PMID- 12560710 TI - Reversed saphenous neurocutaneous island flap: clinical experience and evolution to the posterior tibial perforator-saphenous subcutaneous flap. PMID- 12560711 TI - Distally based sural island flap. PMID- 12560712 TI - Acute adenosine treatment is effective in augmentation of ischemic tolerance in muscle flaps in the pig: an update. PMID- 12560713 TI - Endothelial progenitor cells: the promise of vascular stem cells for plastic surgery. PMID- 12560714 TI - Direct and indirect perforator flaps: the history and the controversy. AB - LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Recognize the major role of the vascular supply to a cutaneous flap. 2. Predict its reliability. 3. Understand basic schemes for classification. 4. Realize that the evolution of these concepts is an ongoing dynamic process. Currently, the vascular supply to the fascial plexus is considered the factor of greatest importance in ensuring the reliability of any skin-bearing flap. The multiplicity of origins of the deep fascial perforators to this plexus has led to a bewildering array of terminology intended to encompass all possible flap options. A brief review of the history of the evolution of cutaneous flaps provides insight essential in understanding a simple proposal for their classification. Because all fascial perforators course either directly from a source vessel or indirectly first through some other tissue to ultimately reach the suprafascial layer, the corresponding flaps based on any such perforators could most simply be termed either direct perforator flaps or indirect perforator flaps, respectively. PMID- 12560715 TI - Botulinum toxin type a: once feared, now revered. PMID- 12560716 TI - Reinforced orbitotemporal lift: contribution to midface rejuvenation. AB - The changes in the aging face occur from progressive ptosis of the skin, fat, and muscle, in conjunction with bone absorption and cartilage atrophy. In the orbital region, hollowness and compartmentalization occur. Conventional face lift procedures correct only the skin flaccidity, and superficial musculoaponeurotic system techniques reposition the skin and platysma without repositioning the middle third of the face, creating an artificial jawline. Subperiosteal rhytidectomy disrupts the anatomy of the periorbita, which gives the patient a certain scarecrow aspect. Composite rhytidectomy associated with brow lift and blepharoplasty may offer better results, with improvement in the malar and orbital regions. The reinforced orbitotemporal lift (ROTEL) is a new procedure in a face lift that allows the orbicularis oculi muscle and all the structures connected to it to be elevated and stretched and the orbitotemporal skin to be raised, repositioning these structures and ending orbital compartmentalization. The result is an impressive improvement in the malar-orbitotemporal region, resulting in a natural and youthful appearance. PMID- 12560717 TI - A new resorbable tack fixation technique for endoscopic brow lifts. AB - The endoscopic brow lift is now widely accepted in aesthetic plastic surgery, and various fixation techniques have been described in the literature. New developments and technology have expanded the use of resorbable devices in different surgical specialties, including plastic surgery. The authors present a technique that offers simple, fast, and reliable forehead fixation for endoscopic brow lifts using resorbable tacks. Successful facial rejuvenation was obtained in the majority of the patients without complications, need for follow-up visits to tighten the flap fixation system, or secondary procedures to extract the fixation system. PMID- 12560718 TI - Ear reconstruction after auricular chondritis secondary to ear piercing. AB - The recent fad of high ear piercing in the pinna has led to an increased incidence of auricular chondritis, which leads to dissolution of the cartilage and residual ear deformity. The typical postpiercing chondritis deformity presents as a structural collapse of the superior helical rim, scaphal cartilage, and the adjacent antihelix. The skin envelope is usually preserved, but it may be scarred from the infectious process and from previous drainage incisions. In the present article, the authors present a systematic approach to reconstruction of these acquired ear deformities. Careful assessment of the residual tissue is requisite to planning and appropriate reconstruction. The greater the cartilage loss, the more structural support is required to expand the skin envelope to its normal size and shape. The choice of cartilage donor site is made on the basis of the size of the defect and may include ipsilateral or contralateral conchal cartilage, bilateral conchal cartilage, or costal cartilage. Redraping of the carefully dissected skin and fixation of the flaps to the newly reconstructed cartilaginous framework usually provide sufficient soft-tissue coverage. A temporal-parietal fascial flap is preserved for the rare cases of extensive full thickness skin loss or badly damaged and scarred auricular skin. PMID- 12560719 TI - Chin augmentation with conchal cartilage. AB - Profile enhancement most frequently involves mentoplasty. For this purpose, the author performs conchal grafts to the chin. A total of 28 women aged 15 to 76 years (mean age, 38.11 +/- 15.11 years) requested mentoplasty by itself or combined with rhinoplasty, rhytidoplasty, or submental lipoplasty. The conchal cartilage was harvested subperichondrally through a posterior 3-cm incision. The specimen measured 3 x 1 cm, which was sufficient to project the chin 2 to 3 mm. For 4 to 5 mm of projection, both cartilages were used. The graft was positioned under the periosteum and held with two 5-0 nylon sutures. In these cases, the conchal cartilage graft was a suitable option for chin augmentations up to 5 mm. PMID- 12560720 TI - Subfascial breast implant: a new procedure. AB - Breast augmentation techniques using the submuscular and subglandular planes to introduce the implant are well known and widely used procedures. The authors have been using the subfascial dissection plane, a new concept for breast augmentation. From October of 1998 to September of 2001, 263 patients underwent breast augmentation. In all cases, the implants were inserted into the subfascial plane. McGhan 410 anatomic biodimensional, cohesive gel implants (size, 155 to 310 g) were used. There are additional benefits using this technique; these include avoiding implant deformation or distortion (as seen in the retromuscular position), leaving additional soft tissue between the implant and the skin, and minimizing implant edge prominence (inherent to retroglandular placement). These technical details lead to fewer patient complaints. Morbidity is similar to that of other techniques. PMID- 12560721 TI - Classification and management of gynecomastia: defining the role of ultrasound assisted liposuction. AB - Gynecomastia, or excessive male breast development, has an incidence of 32 to 65 percent in the male population. This condition has important physical and psychological impacts. Advances in elucidating the pathophysiology of gynecomastia have been made, though understanding remains limited. Recommendations for evaluation and workup have varied and are often arbitrary. A diagnostic algorithm is suggested, with emphasis on a comprehensive history, physical examination, and minimizing unnecessary diagnostic testing. Medical management has had limited success; surgical therapy, primarily through excisional techniques, has been the accepted standard. Although effective, excisional techniques subject patients to large, visible scars. Ultrasound assisted liposuction has recently emerged as a safe and effective method for the treatment of gynecomastia. It is particularly efficient in the removal of the dense, fibrous male breast tissue while offering advantages in minimal external scarring. A new system of classification and graduated treatment is proposed, based on glandular versus fibrous hypertrophy and degree of breast ptosis (skin excess). The authors' series of 61 patients with gynecomastia from 1987 to 2000 at the University of Texas Southwestern Department of Plastic Surgery demonstrated an overall success rate of 86.9 percent using suction-assisted lipectomy (1987 to 1997) and ultrasound-assisted liposuction (1997 to 2000). The authors have found ultrasound-assisted liposuction to be effective in treating most grades of gynecomastia. Excisional techniques are reserved for severe gynecomastia with significant skin excess after attempted ultrasound-assisted liposuction. PMID- 12560722 TI - An update on the role of subcutaneous infiltration in suction-assisted lipoplasty. PMID- 12560723 TI - The Ptolemaic concept versus the Copernican system: rethinking the etiology and results of treatment in craniosynostosis in 2002. PMID- 12560724 TI - Get a life or two or more! PMID- 12560725 TI - Anti-aging medicine: part I. Hormone replacement therapy in women. PMID- 12560726 TI - Beyond survival: establishment of a breast reconstruction support group. PMID- 12560727 TI - The use of tissue expander base as an internal nasal splint. PMID- 12560728 TI - Hyperhidrosis. PMID- 12560729 TI - A clinical definition of an ideal nasal radix. PMID- 12560730 TI - The propanolol-epinephrine interaction revisited: a serious and potentially catastrophic adverse drug interaction in facial plastic surgery. PMID- 12560731 TI - Large-volume tumescent anesthesia for extensive liposuction in oriental patients: lidocaine toxicity and its safe dose level. PMID- 12560732 TI - Skin marking in plastic surgery: a helpful suggestion. PMID- 12560733 TI - The ideal patient for the first breast reconstruction using a diep flap. PMID- 12560734 TI - Tram flap delay: which vessels should be ligated? PMID- 12560735 TI - Treatment of hemangiomas in children. PMID- 12560736 TI - Disseminated basal cell carcinoma. PMID- 12560737 TI - A simple technique for reduction gonioplasty. PMID- 12560738 TI - Treatment of severe neurofibromatosis of the nipple-areola complex. PMID- 12560739 TI - Curve analysis of the aging orbital aperture. PMID- 12560740 TI - A role for the anesthesiologist in elective cosmetic surgery? PMID- 12560741 TI - The electrocardiography dot as a preoperative marker for nipple-areola complex reconstruction. PMID- 12560742 TI - Gestational gigantomastia after reduction mammaplasty: complication or coincidence? PMID- 12560743 TI - Suction-curettage: treatment for axillary hyperhidrosis and hidradenitis. PMID- 12560744 TI - Complete absence of breasts in a 24-year-old woman associated with ectodermal dysplasia. PMID- 12560745 TI - A simple method of harvesting a thin iliac bone graft for reconstruction of the orbital wall. PMID- 12560746 TI - Torpid ulcer on the achilles tendon: conservative treatment with stanozolol and pinch grafting. PMID- 12560747 TI - The small-wave incision for long keloids. PMID- 12560748 TI - Reconstruction of maxilla with free latissimus dorsi-scapular osteomusculocutaneous flap. PMID- 12560749 TI - A useful application of the double-breasted vest principle in skin closure. PMID- 12560750 TI - The effect of facial muscle surgery on migraine. PMID- 12560752 TI - CD40 in hepatocellular carcinoma: relevant or not? AB - The expression of the CD40 receptor in different tumours suggests that it might be involved in tumour development and proliferation. In this article, we discuss recent findings on CD40 expression in hepatocellular carcinoma, its possible biological role for tumour development, and different experimental therapeutic strategies such as gene therapy and immunotherapy protocols. PMID- 12560753 TI - Enteral nutrition in Crohn's disease: fat in the formula. AB - Enteral nutrition is effective in inducing remission in active Crohn's disease. Speculation on the underlying mechanism of action has moved away from the presentation of nitrogen and towards the fat content of the various enteral feeds. Evidence is accumulating that additional long-chain triglyceride in such feeds impairs the response rate in active Crohn's disease, whereas no deleterious effects of additional medium-chain triglyceride have been identified. It has been proposed that long-chain triglycerides composed from n-6 fatty acids may be the most harmful, since such fatty acids are substrates for inflammatory eicosanoid production. However, recent studies comparing different enteral feeds are not consistent in identifying which additional fatty acids impair response rates to the greatest extent. Despite meta-analyses concluding that polymeric diets (typically containing large amounts of fat) are as effective as elemental diets, it would seem sensible to use enteral feeds with minimal fat content when treating active Crohn's disease. PMID- 12560754 TI - Heterogeneous expression and regulation of CD40 in human hepatocellular carcinoma. AB - OBJECTIVE: CD40, a member of the tumour necrosis factor receptor family, plays a major role in adaptive immune responses and contributes to cancer surveillance. Conflicting results have been reported recently on the expression and function of CD40 in carcinomas. The aim of the present study was to investigate the role of CD40 in human hepatoma. DESIGN/METHODS: CD40 expression was examined in hepatomas and derived cell lines by immunohistochemistry, flow cytometry and reverse transcriptase polymerase chain reaction. We investigated in hepatoma cell lines the regulation of CD40 by pro-inflammatory cytokines and the effects of its ligation with soluble CD40L on the expression of co-stimulatory and pro-apoptotic cell-surface molecules and survival. RESULTS: CD40 was detected with a similar frequency of about 40% in hepatoma specimens and derived cell lines but not in normal hepatocytes. Tumour necrosis factor alpha and its combination with interferon gamma upregulated CD40 only in intrinsically positive cell lines. CD40 ligation had no effect on cell viability or surface expression of CD54, CD80, CD86 or CD95. CONCLUSIONS: CD40 is expressed variably in human hepatoma and enhanced by distinct pro-inflammatory cytokines. The lack of detectable effects of CD40 ligation does not support a major role of this molecule in hepatocellular carcinoma biology. PMID- 12560755 TI - Activin A and follistatin in acute liver failure. AB - BACKGROUND: Liver regeneration may be impaired in acute liver failure due to either inhibition of the proliferative response or ongoing liver cell death. Activin A, a member of the TGFbeta superfamily, inhibits hepatocyte DNA synthesis and induces apoptosis. METHODS: Levels of activin A and its binding protein follistatin in the serum of 23 patients with acute liver failure were determined by enzyme-linked immunosorbent assay. RESULTS: Serum activin A was significantly increased in acute liver failure patients (median 2.15 ng/ml, range 0.28-6.87 ng/ml) compared to normal controls (median 0.25 ng/ml, range 0.19-0.53 ng/ml; = 10; 0.001). However, this was not linked to the final disease outcome. Higher levels of activin A were found in the serum of patients with acute liver failure due to paracetamol overdose (median 2.87 ng/ml, range 0.72-6.87 ng/ml; = 17) than in patients with acute liver failure due to non-A to E hepatitis (median 1.10 ng/ml, range 0.28-2.70 ng/ml; = 6; 0.05). Serum follistatin was also increased in acute liver failure patients (median 2.84 ng/ml, range 0.57-13.24 ng/ml) compared to normal controls (median 0.68 ng/ml, range 0.32-3.70 ng/ml; 0.01). CONCLUSION: Serum activin A is increased in acute liver failure and could be a factor in the inhibition of liver regeneration. PMID- 12560756 TI - Viraemia, cryoglobulins and autoantibodies in haemodialysis patients infected with hepatitis C virus. AB - OBJECTIVES: The clinical features of hepatitis C virus (HCV) infection depend on the immune and autoimmune reactions induced by the virus. Chronic renal failure might alter the pattern of these reactions. The aim of this study was to determine the prevalence of cryoglobulinaemia, the frequency of autoantibodies and HCV viral load in HCV infected Greek patients on chronic haemodialysis. METHODS: Seventy-three HCV Ab(+) patients on maintenance haemodialysis and 87 otherwise normal patients with chronic HCV infection were evaluated for the presence of cryoglobulins, autoantibodies and viral markers. RESULTS: Cryoglobulins were detected in 22/73 (30.1%) haemodialysis patients and in 23/87 (26.4%) patients with normal renal function (NS). The mean cryocrit value was significantly lower in the haemodialysis group ( = 0.002). Haemodialysis patients had significantly higher levels of C4 component of complement and lower incidence of rheumatoid factor than those of patients with normal renal function. Serum HCV RNA levels were found significantly lower in the haemodialysis group (median, 2.20 Meq/ml; range, 119.9 Meq/ml) than in the group with normal renal function (median, 4.50 Meq/ml; range, 114.9 Meq/ml; = 0.046). The distribution of genotypes was not different between the two groups. CONCLUSIONS: There are subtle differences in autoimmune features of HCV infection if the patients are also haemodialysed for renal failure. HCV viral load is lower in haemodialysis patients, with no difference in the HCV genotype prevalence. The clinical significance of these findings is unknown. PMID- 12560757 TI - Proliferation of antigen MIB-1 in metastatic carcinoid tumours removed at liver transplantation: relevance to prognosis. AB - BACKGROUND: Metastatic carcinoid tumours are difficult to manage. In spite of a multidisciplinary approach, including orthotopic liver transplantation, the recurrence rate is high with a poor prognosis. Histopathology generally fails to provide prognostic information, hence it is essential to try to identify markers of prognosis in these tumours before considering orthotopic liver transplantation. The MIB-1 antibody, which detects cell proliferative activity, has been shown to be a useful prognostic marker for a variety of neoplasms. AIMS: To assess the value of MIB-1 immunostaining as a prognostic marker of the duration to recurrence and the survival of patients undergoing orthotopic liver transplantation for metastatic carcinoid/neuroendocrine tumours of the liver. METHODS: Fourteen patients were included in the study. Formalin-fixed, paraffin embedded tissue sections of the tumours were stained with routine haematoxylin and eosin and chromogranin. The cell proliferative activity was assessed by MIB-1 antibody labelling using the immunoperoxidase method. Results were correlated with the time of tumour recurrence and the length of patients' survival after transplantation. RESULTS: No correlation was found between MIB-1 labelling index and age, gender, clinical and histological type of tumour (i.e. carcinoid, APUDOMA, secreting or non-secreting). The patients with higher MIB-1 indices ( 5%) showed a trend toward earlier recurrence and poorer survival than those with low MIB-1 indices ( 5%). The predictive value of a MIB-1 index of 2 indicating patient survival of 24 months was 83% (five out of six patients). CONCLUSIONS: The correlation between MIB-1 index and patients' survival suggests that a high proliferative rate, as assessed by MIB-1 immunostaining, may detect those tumours with more aggressive biological behaviour. Prospective studies on a larger number of patients will be needed to determine if, in any individual tumour, this method will provide an additional parameter for a rational approach to therapy. PMID- 12560758 TI - Intestinal mucosal oxidative damage and bacterial translocation in cirrhotic rats. AB - BACKGROUND: Bacterial translocation plays an important role in the pathogenesis of spontaneous bacterial peritonitis mainly due to intestinal bacterial overgrowth. Alterations in the functional integrity of the intestinal barrier caused by an increased production of free radical metabolites as a consequence of portal hypertension could also facilitate bacterial translocation in cirrhotic rats. OBJECTIVE: The aim of the study was to determine intestinal mucosal lipid peroxidation and neutrophil infiltration and their relationship with portal hypertension and bacterial translocation in cirrhotic rats. DESIGN: Eighteen male Sprague-Dawley rats with cirrhosis induced by carbon tetrachloride, administered by gavage, and eight control rats were included in the study. METHODS: Samples of jejunum, ileum and caecum were obtained by laparotomy for the determination of malondialdehyde and myeloperoxidase as indexes of lipid peroxidation and neutrophil infiltration, respectively. Samples of ascitic and pleural fluids, mesenteric lymph nodes and ileal stools were obtained for the culture of microoganisms. RESULTS: The concentration of malondialdehyde was significantly higher in ileal and caecal, but not in jejunal mucosa, in cirrhotic rats, mainly in those with ascites (P< 0.01), as compared to control rats (P< 0.01), and in cirrhotic rats with bacterial translocation compared to those without bacterial translocation (P< 0.01). No differences between groups were observed in the concentrations of myeloperoxidase in jejunum, ileum or caecum. A direct correlation between ileal malondialdehyde and portal pressure was observed (P< 0.01). CONCLUSIONS: Cirrhotic rats, particularly those with ascites and bacterial translocation, show increased malondialdehyde levels in ileal and caecal mucosa. These results suggest that mucosal oxidative damage in ileum and caecum could favour bacterial translocation in cirrhotic rats. PMID- 12560759 TI - Dietary fat attenuates the benefits of an elemental diet in active Crohn's disease: a randomized, controlled trial. AB - OBJECTIVES: Although an elemental diet has been established as the primary treatment for patients with Crohn's disease, the influence of dietary fat on the elemental diet remains unclear. We have designed the first randomized, controlled trial for elemental diets containing different fat percentages in patients with active Crohn's disease. METHODS: Each patient was randomized to receive one of three dose levels of fat in an elemental diet (Elental) for 4 weeks: 10 patients received low fat (3.06 g/day), 10 patients received medium fat (16.56 g/day) and eight patients received high fat (30.06 g/day). The additional fat was composed of long-chain fatty acids. All patients were evaluated using the International Organization of Inflammatory Bowel Disease rating, plus C-reactive protein level and erythrocyte sedimentation rate, which were measured at weekly intervals. RESULTS: Although the International Organization of Inflammatory Bowel Disease rating, C-reactive protein level and erythrocyte sedimentation rate in the low fat group decreased, the values in the medium- and high-fat groups fluctuated during the study. The remission rate after 4 weeks in each group was 80%, 40% and 25% for patients in the low-, medium- and high-fat groups, respectively. CONCLUSIONS: When the fat consisted of long-chain triglycerides, a high amount of this fat in the elemental diet formula decreased its therapeutic effect against active Crohn's disease. PMID- 12560760 TI - Association of coeliac disease with primary biliary cirrhosis in Poland. AB - OBJECTIVES: In western and northern but not southern Europe, the prevalence of coeliac disease among patients with primary biliary cirrhosis (PBC) is higher than in the general population. We analysed the prevalence of coeliac disease among patients with PBC in Poland, a central European country. METHODS: In 115 patients with PBC, immunoglobulin A (IgA) antibodies against guinea-pig tissue transglutaminase (tTGA), monkey endomysium (EMA) and gliadin (AGA) were determined. In patients positive for tTGA and/or EMA, the DNA typing of HLA-DQB gene and small-bowel biopsy were performed. RESULTS: IgA EMA was found in one patient with PBC (0.9%, 95% CI 0-2.5%). tTGA was detected in seven patients (6%, 95% CI 1.8-10.3%). Small-bowel biopsy showed flat mucosa in one subject, who was EMA positive/tTGA negative/AGA negative, and normal histology in four tTGA positive/EMA-negative patients. In the latter four patients, the positive tTGA result was caused by IgA reactivity to proteins other than transglutaminase. Prevalence of coeliac disease in our 115 patients with PBC was 0.9% (95% CI 0 1.9%). In one patient with silent coeliac disease presenting with the HLA-DQ2 allele, introduction of a gluten-free diet was followed by improvement in liver function tests, improvement in histology of the distal duodenum, and disappearance of EMA. CONCLUSIONS: Our results from Poland do not confirm a high prevalence of coeliac disease in PBC patients. Guinea-pig liver transglutaminase immunolinked assay cannot be used as a screening test for coeliac disease in PBC patients. A gluten-free diet may be helpful in restoration of liver function in patients with such an association. PMID- 12560761 TI - Irritable bowel syndrome subtypes according to bowel habit: revisiting the alternating subtype. AB - BACKGROUND: Disturbed bowel habit, diarrhoea or constipation is a key manifestation of irritable bowel syndrome (IBS). In some patients, diarrhoea and constipation alternate, giving rise to the so-called alternating subtype. AIMS: To assess IBS subtype breakdown (constipation (C-IBS), diarrhoea (D-IBS) or alternating (A-IBS)) according to the Rome II criteria and patients' self assessment, the predominance in the alternating subtype (i.e. constipation, diarrhoea or neither), and the medical and personal impact, including health related quality of life (HRQoL), of the different IBS subtypes. SUBJECTS AND METHODS: Two thousand individuals selected randomly to represent the general population were classified as potential IBS subjects (n = 281) or as non potential IBS subjects (n = 1719) according to a validated questionnaire. Bowel habit classification was determined using the Rome II IBS supportive symptoms. RESULTS: Among 201 subjects meeting the Rome I criteria, 15% presented with D IBS, 44% presented with C-IBS, 19% presented with A-IBS, and 22% presented with normal bowel habit. Among the 63 subjects meeting the Rome II criteria, 23% presented with A-IBS. According to the subjects' self-assessment, of those meeting the Rome I criteria, 16% considered themselves to have D-IBS, 66% to have C-IBS and 18% to have A-IBS. In subjects meeting the Rome II criteria, 24% considered themselves to have A-IBS. Among those classified with A-IBS by the Rome II criteria, most considered themselves to be constipated. Regardless of the subtype self-classification, most subjects reported a normal frequency of bowel movements. Clinical manifestations in A-IBS were very similar to those of C-IBS but with the added presence of defecatory urgency. Abdominal discomfort/pain and frequency of visits to physicians were greater in the A-IBS subtype than in the other two IBS subtypes. HRQoL was affected similarly in all IBS subtypes. CONCLUSIONS: Approximately one-quarter of subjects with IBS belong to the A-IBS subtype by the Rome II criteria, although the majority consider themselves to be constipated; indeed, clinical manifestations are more akin to the C-IBS subtype than to the D-IBS subtype. Abdominal discomfort/pain and frequency of visits to physicians are greater in the A-IBS subtype than in the other two IBS subtypes, while HRQoL is impaired similarly. PMID- 12560762 TI - Risk of upper gastrointestinal bleeding associated with non-aspirin cardiovascular drugs, analgesics and nonsteroidal anti-inflammatory drugs. AB - OBJECTIVE: To evaluate the risk of upper gastrointestinal bleeding associated with non-aspirin cardiovascular drug therapy, common analgesics and individual nonsteroidal anti-inflammatory drugs (NSAIDs). METHODS: The case group was made up of 1122 consecutive patients admitted with bleeding from a peptic lesion. The 2231 control subjects consisted of 1109 patients hospitalized for other reasons and 1122 outpatients from the same geographical area. The relative risk was calculated by unconditional logistic regression after adjusting for confounding factors. RESULTS: The use of the antiplatelet agent triflusal, and other commonly used cardiovascular drugs, such as beta-receptor blockers and calcium channel blockers, was not associated with increased risk of upper gastrointestinal bleeding. The use of angiotensin-converting enzyme inhibitors reduced the risk of bleeding by 30% (odds ratio 0.7; 95% confidence interval 0.5-0.96). Use of ketorolac (odds ratio 59.4; 95% confidence interval 7.7-454) and piroxicam (odds ratio 19.6; 95% confidence interval 9.3-35.3) carried the highest risk. Use of paracetamol and tramadol was not associated with increased risk of bleeding, but the non-narcotic agent metamizol was associated with a small increase in risk of upper gastrointestinal bleeding (odds ratio 2.6; 95% confidence interval 1.3 5.2). CONCLUSIONS: The use of the antiplatelet agent triflusal and other cardiovascular drugs apart from low-dose aspirin was not associated with gastrointestinal bleeding. The use of either NSAIDs or aspirin increased the risk of gastrointestinal bleeding but, among the analgesics, only metamizol induced a small increase in the risk of gastrointestinal bleeding. PMID- 12560763 TI - Helicobacter pylori infection in Swiss adolescents: prevalence and risk factors. AB - OBJECTIVE: This investigation sought to study the influence exerted by demographic and socioeconomic factors on the prevalence of infection in adolescents living in Switzerland. DESIGN: Epidemiological study spanning 3 years. METHODS: We included 196 15- to 16-year-old adolescents from a north eastern Swiss city in our study, recruited by the school health service during a medical check-up in the years 1999, 2000 and 2001. infection was detected by ELISA using 2nd generation anti- IgG antibodies. Demographic and socioeconomic data were collected by questionnaire. RESULTS: infection was found in 19 of the 196 (9.7%) tested adolescents. tested positive in, respectively, 13 (7.3%) of the 176 natives and six (30%) of the 20 (P = 0.01 chi-squared) subjects from foreign countries. infection was significantly highly correlated with demographic factors but did not correlate with most of the socioeconomic factors. CONCLUSION: The rate of infection among Swiss adolescents is one of the lowest in Europe. Nevertheless, an important disparity is evident between the rate of infection observed in the native population and that among immigrants. High living standards available to the majority of the population may explain the minor influence of socioeconomic factors on infection in our country. PMID- 12560764 TI - Neoplasia of the tongue in a patient with Crohn's disease treated with azathioprine: case report. AB - Azathioprine is now widely used for the maintenance treatment of Crohn's disease, but there are concerns whether azathioprine could predispose to malignancy in patients with inflammatory bowel disease. We report here a case of a 39-year-old non-smoking male with Crohn's disease who had been treated for 3 years with azathioprine and developed a lingual ulcer. Biopsy revealed squamous cell carcinoma, a tumour not previously associated with Crohn's disease. PMID- 12560765 TI - Alternative approach to massive gastrointestinal bleeding in ulcerative colitis: highly selective transcatheter embolization. AB - Massive haemorrhage in ulcerative colitis mainly occurs as a result of exacerbated disease, usually acute or even fulminant colitis. Emergency proctocolectomy is currently advocated as the only reliable treatment of this patient group. However, this type of surgical intervention has a high rate of morbidity and mortality. Recently, several promising studies have been published that describe transcatheter embolization for the treatment of massive lower gastrointestinal bleeding in cases of bleeding colonic diverticular disease and angiodysplasia. Success rates of 48-88% have been reported. To avoid the high risk associated with emergency colectomy, this modern procedure was performed in an ulcerative colitis patient with intractable lower gastrointestinal bleeding. To our knowledge, this is the first patient with ulcerative colitis presenting with massive lower gastrointestinal blood loss to be successfully treated by highly selective transcatheter embolization. This interventional procedure is suggested as an alternative therapeutic approach to haematochezia in selected ulcerative colitis patients. PMID- 12560766 TI - Small rectal carcinoid with lymph node metastasis diagnosed prior to treatment. AB - We treated a 59-year-old man with a small, yellowish, submucosal rectal tumour that was detected incidentally during a colonoscopic examination. Endoscopic ultrasonography revealed a hypoechoic submucosal tumour 13 mm in diameter. Computed tomography and magnetic resonance imaging studies were performed, and pararectal and para-obturator lymph node involvement was confirmed. The patient underwent Miles' operation with lymph node dissection; 24 months later, he is disease free. We were able to diagnose rectal carcinoid tumour and evaluate the lymph node metastasis prior to surgery. Even with a small carcinoid, it is important to determine the depth of invasion and the presence of lymph node metastasis prior to treatment. PMID- 12560767 TI - Congenital oesophageal stenosis: an atypical presentation in a young woman. AB - Congenital oesophageal stenosis is a very rare disease that commonly occurs in infancy with male predominance. This report describes a highly unusual case of congenital oesophageal stenosis extended throughout the length of the oesophagus, without webs or tracheobronchial remnants in the oesophageal wall, with normal oesophageal motility and normal endosonography in an adult female. PMID- 12560768 TI - Necrotizing enteritis with hepatic portal venous gas and pneumatosis intestinalis: report of a case. AB - A 73-year-old woman visited our hospital because of increasing abdominal distension and lower abdominal pain. On abdominal computed tomography (CT), hepatic portal venous gas (HPVG) and pneumatosis intestinalis of the small intestine were found. HPVG caused by intestinal necrosis was diagnosed, and an emergency laparotomy was thus performed. Necrosis of the small intestine over a 40-cm area from the ileocaecal region toward the mouth was found, and the lesion was resected. Histopathologically, haemorrhagic necrotic enteritis was diagnosed. The patient is alive as of the seventieth day after operation. The prognosis of intestinal necrosis accompanied by HPVG and pneumatosis intestinalis is poor. The presence of HPVG suggests the occurrence of a serious lesion in the abdominal cavity. Therefore, appropriate treatment should be performed immediately. PMID- 12560769 TI - Acute onset dysphagia associated with an intramural oesophageal haematoma in acquired haemophilia. AB - A 78-year-old man presented with a 5-day history of epistaxis and spontaneous bruising, and a 2-day history of acute dysphagia. Barium swallow, computerized tomography scan of the chest and upper gastrointestinal endoscopy were suggestive of an upper oesophageal tumour, although biopsies failed to confirm this. Investigations including a raised activated partial thromboplastin time led to the detection of an inhibitor causing functional factor VIII deficiency. Following treatment with intravenous human immunoglobulin, oral prednisolone and oral cyclophosphamide, the patient's dysphagia resolved. There was a resolution of the findings seen at the initial endoscopy and on computerized tomography scan of the chest, consistent with an oesophageal haematoma. Follow-up endoscopy failed to detect recurrence or an aetiological factor. PMID- 12560770 TI - Multicenter Canadian study of prehospital trauma care. AB - OBJECTIVE: To evaluate whether the type of on-site care a trauma patient receives affects outcome. SUMMARY BACKGROUND DATA: The controversy regarding the prehospital care of trauma patients between Advanced Life Support (ALS) and Basic Life Support (BLS) is ongoing. Due to this unresolved controversy, as well as historical, cultural, and political factors, there are significant variations with respect to the type of prehospital care available for trauma patients. METHODS: This prospective cohort study compared three types of prehospital trauma care systems: Montreal, where physicians provide ALS (MD-ALS); Toronto, where paramedics provide ALS (PMD-ALS); and Quebec City, where emergency medical technicians provide BLS only (EMT-BLS). The study took advantage of this variation to evaluate the association between the type of on-site care and mortality in patients with major life-threatening injuries. All patients were treated at highly specialized tertiary (level I) trauma hospitals. The main outcome measure was death as a result of injury. Follow-up was to hospital discharge. RESULTS: The overall mortality rates by type of on-site personnel were physicians 35%, paramedics 24%, and EMTs 18%. For patients with major but survivable trauma, the overall mortality rates were physicians 32%, paramedics 28%, and EMTs 26%. The overall mortality rate of patients receiving only BLS at the scene was 18% compared to 29% for patients receiving ALS. For the subgroup of patients with major but survivable injuries, the mortality rates were 30% for ALS and 26% for BLS. The adjusted increased risk for mortality in patients receiving ALS at the scene was 21%. CONCLUSIONS: In urban centers with highly specialized level I trauma centers, there is no benefit in having on-site ALS for the prehospital management of trauma patients. PMID- 12560771 TI - Prehospital care: do no harm? PMID- 12560772 TI - Survival of lymph node-negative breast cancer patients in relation to number of lymph nodes examined. AB - OBJECTIVE: To assess the relationship between number of lymph nodes examined and survival of patients diagnosed with node-negative localized breast cancer using a large sample of patients from population-based cancer registries in the United States. SUMMARY BACKGROUND DATA: Conflicting results have been reported from studies on the relationship between number of lymph nodes examined and survival of patients diagnosed with node-negative localized breast cancer. METHODS: The study included 69,543 patients diagnosed in 1988-97 with localized invasive node negative breast cancer reported to nine population-based registries in the U.S. National Cancer Institute's Surveillance, Epidemiology and End Results (SEER) Program of population-based cancer registries. Hazard ratios for groups defined by number of nodes examined were analyzed in Cox proportional hazards regression models that included age, tumor size and grade, race/ethnicity, and other variables. RESULTS: A significantly higher risk of death from breast cancer was found among patients with 0, 1 to 3, or 4 to 10 nodes examined than with 20-plus nodes examined, even among patients with tumors 2 cm or smaller. CONCLUSIONS: Future studies of survival of node-negative patients, by number of nodes examined, should include information on comorbidity and treatment. PMID- 12560773 TI - Failure of routine axillary nodal sampling to predict survival outcomes in lymph node-negative (N0) breast cancer. PMID- 12560774 TI - Percutaneous local ablative therapy for hepatocellular carcinoma: a review and look into the future. AB - OBJECTIVE: To review and compare treatment result for percutaneous local ablative therapy (PLAT) with surgical resection in the treatment of small hepatocellular carcinoma (HCC). SUMMARY BACKGROUND DATA: PLAT is indicated for small unresectable HCC localized to the liver. From the use of ethanol to the latest technology of radiofrequency ablation, ablative techniques have been refined and their role in the management of HCC established. This review aims to give an overview of various ablative methods, including their efficacy, indications, and limitations, and also tries to look into the future of clinical trials in PLAT. METHODS: The authors reviewed recent papers in the English medical literature about the use of local ablative therapy for HCC. Focus was given to the results of treatment in terms of local control, progression-free survival, and overall survival, and to compare treatment results with those of surgery. RESULTS: PLAT for small HCC (<5 cm) with thermal ablation (radiofrequency ablation or microwave coagulation) can achieve effective local control of disease and is superior to ethanol injection. Progressive disease in untreated areas is a common reason for failure. Overall progression-free survival is similar to that of surgical resection. CONCLUSIONS: Thermal ablation gives good local control of small HCC, is superior to ethanol, and may be comparable to surgical resection in long-term outcome. PMID- 12560775 TI - Vein reconstruction in modified right liver graft for living donor liver transplantation. AB - OBJECTIVE: To report the authors' experience with hepatic vein reconstruction and plasty in living donor liver transplantation for adult patients. SUMMARY BACKGROUND DATA: A right liver graft without the middle hepatic vein (MHV) trunk (modified right liver graft) can cause severe congestion of the right paramedian sector. However, the need for MHV reconstruction has not been fully recognized. METHODS: From June 2000 to December 2001, 30 adult patients received a modified right liver graft. Major MHV tributaries were preserved and reconstructed under the authors' criteria. Plasty of recipient hepatic veins for a wide outflow orifice was performed when necessitated. The regeneration of paramedian and lateral sectors of the grafts was examined by computed tomography 1 and 3 months after the operation. RESULTS: MHV tributaries were reconstructed in 18 grafts. Plasty of recipient hepatic veins was performed in 15 patients. All patients survived the operation. The regeneration of paramedian and lateral sectors was equivalent. CONCLUSIONS: A modified right liver graft can provide satisfactory surgical results if hepatic vein reconstruction and plasty are performed using the present techniques. PMID- 12560776 TI - How safe is high-power ultrasonic dissection? AB - OBJECTIVE: To evaluate the safety of ultrasonic dissection. SUMMARY BACKGROUND DATA: High-power ultrasonic dissection is in widespread use for both open and laparoscopic operations and is generally perceived to carry a low risk of collateral damage, but there is no published evidence for this. METHODS: Under controlled experimental conditions, ultrasonic dissections were performed in pigs using Ultracision (Ethicon) or Autosonix (Tyco/USSC) at the three power settings (3, 4, and 5) in random fashion to mobilize the cardia and fundus, bile duct, hepatic artery, portal vein, aorta from the inferior vena cava, renal vessels, colon, and ureters. The dissections (open and laparoscopic) were carried out on pigs at each power setting with each device. Thermal mapping of the tissues during dissection was performed with an infrared thermal camera and associated software. The animals were killed at the end of each experiment and specimens were harvested for quantitative histology. RESULTS: Extreme and equivalent temperature gradients were generated by ultrasonic dissection with both systems. Heat production was directly proportional to the power setting and the activation time. The core body temperature of the animals after completion of the laparoscopic dissections rose by an average of 2.3 degrees C. The zone around the jaws that exceeded 60 degrees C with continuous ultrasonic dissection for 10 to 15 seconds at level 5 measured 25.3 and 25.7 mm for Ultracision and Autosonix, respectively. At this power setting and an activation time of 15 seconds, the temperature 1.0 cm away from the tips of the instrument exceeded 140 degrees C. Although there was no discernible macroscopic damage, these thermal changes were accompanied by significant histologic injury that extended to the media of large vessels and caused partial- to full-thickness mural damage of the cardia, ureter, and bile duct. Collateral damage was absent or insignificant after dissections at power level 3 with both systems and an activation time not exceeding 5 seconds. CONCLUSIONS: High-power ultrasonic dissections at level 5 and to a lesser extent level 4 result in considerable heat production that causes proximity collateral damage to adjacent tissues when the continuous activation time exceeds 10 seconds. Ultrasonic dissections near important structures should be conducted at level 3. At power levels of 4 and 5, the ultrasonic energy bursts to the tissue should not exceed 5 seconds at any one time. PMID- 12560777 TI - Canine ileal motor activity after a model of jejunoileal autotransplantation. AB - OBJECTIVE: To determine mechanisms by which extrinsic innervation to the jejunoileum controls ileal motility. SUMMARY BACKGROUND DATA: Small bowel transplantation is complicated by diarrhea and delayed gastric emptying, possibly secondary to altered motility. Ileal motility after small bowel transplantation is poorly characterized. METHODS: Motor activity was recorded from four dogs during fasting and after feeding small (64 Kcal) or large (256 Kcal) meals. Short chain fatty acids known to induce unique ileal motor patterns were administered into the distal ileum during fasting. Dogs were studied before and after jejunoileal denervation simulating autotransplantation. RESULTS: After jejunoileal denervation, the ileal migrating motor complex (MMC) persisted but was no longer temporally coordinated with duodenal MMCs. Spontaneous giant migrating contractions occurred more frequently after denervation and more commonly originated proximally in the jejunum, but the velocity of migration did not differ. In contrast, the incidence and characteristics of spontaneous discrete clustered contractions (DCCs) did not differ. Short-chain fatty acids reproducibly initiated giant migrating contractions and discrete clustered contractions in the distal ileum without differences before and after denervation. Large but not small meals inhibited the ileal MMC after denervation. CONCLUSIONS: Extrinsic innervation and/or intrinsic neural continuity with the duodenum and/or colon control temporal coordination of ileal motility with the duodenum and modulate postprandial inhibition of fasting motility and presence of giant migrating contractions. These changes in motility patterns may prove important in mediating enteric dysfunction after small bowel transplantation. PMID- 12560778 TI - Immunohistochemically demonstrated lymph node micrometastasis and prognosis in patients with otherwise node-negative hilar cholangiocarcinoma. AB - OBJECTIVE: To investigate whether immunohistochemically demonstrated lymph node micrometastasis has prognostic significance in patients with histologically node negative (pN0) hilar cholangiocarcinoma. SUMMARY BACKGROUND DATA: The clinical significance of immunohistochemically detected lymph node micrometastasis recently has been evaluated in various tumors. However, no reports have addressed this issue with regard to hilar cholangiocarcinoma. METHODS: A total of 954 lymph nodes from surgical specimens of 45 patients with histologically node-negative hilar cholangiocarcinoma who underwent macroscopically curative resection were immunostained with monoclonal antibody against cytokeratins 8 and 18. The results were examined for relationships with clinical and pathologic features and with patient survival. RESULTS: Lymph node micrometastases were detected immunohistochemically in 11 (24.4%) of the 45 patients, being found in 13 (1.4%) of 954 lymph nodes examined. Of the 13 nodal micrometastases, 11 (84.6%) were found in the N2 regional lymph node group rather than N1. Clinicopathologic features showed no associations with lymph node micrometastases. Survival curves were essentially similar between patients with and without micrometastasis. In addition, the grade of micrometastasis showed no effect on survival. The Cox proportional hazard model identified microscopic venous invasion, microscopic resection margin status, and histologic differentiation as significant prognostic factors in patients with pN0 disease. CONCLUSIONS: Lymph node micrometastasis has no survival impact in patients with otherwise node-negative hilar cholangiocarcinoma. The authors do not recommend extensive lymph node sectioning with keratin immunostaining for prognostic evaluation. PMID- 12560779 TI - Portal vein embolization before right hepatectomy: prospective clinical trial. AB - OBJECTIVE: To assess the impact of liver hypertrophy of the future liver remnant volume (FLR) induced by preoperative portal vein embolization (PVE) on the immediate postoperative complications after a standardized major liver resection. SUMMARY BACKGROUND DATA: PVE is usually indicated when FLR is estimated to be too small for major liver resection. However, few data exist regarding the exact quantification of sufficient minimal functional hepatic volume required to avoid postoperative complications in both patients with or without chronic liver disease. METHODS: All consecutive patients in whom an elective right hepatectomy was feasible and who fulfilled the inclusion and exclusion criteria between 1998 and 2000 were assigned to have alternatively either immediate surgery or surgery after PVE. Among 55 patients (25 liver metastases, 2 cholangiocarcinoma, and 28 hepatocellular carcinoma), 28 underwent right hepatectomy after PVE and 27 underwent immediate surgery. Twenty-eight patients had chronic liver disease. FLR and estimated rate of functional future liver remnant (%FFLR) volumes were assessed by computed tomography. RESULTS: The mean increase of FLR and %FFLR 4 to 8 weeks after PVE were respectively 44 +/- 19% and 16 +/- 7% for patients with normal liver and 35 +/- 28% and 9 +/- 3% for those with chronic liver disease. All patients with normal liver and 86% with chronic liver disease experienced hypertrophy after PVE. The postoperative course of patients with normal liver who underwent PVE before right hepatectomy was similar to those with immediate surgery. In contrast, PVE in patients with chronic liver disease significantly decreased the incidence of postoperative complications as well as the intensive care unit stay and total hospital stay after right hepatectomy. CONCLUSIONS: Before elective right hepatectomy, the hypertrophy of FLR induced by PVE had no beneficial effect on the postoperative course in patients with normal liver. In contrast, in patients with chronic liver disease, the hypertrophy of the FLR induced by PVE decreased significantly the rate of postoperative complications. PMID- 12560780 TI - High-grade extremity soft tissue sarcomas: factors predictive of local recurrence and its effect on morbidity and mortality. AB - OBJECTIVE: To identify patient characteristics associated with the development of local recurrence and the effect of local recurrence on subsequent morbidity and mortality in patients with intermediate- to high-grade extremity soft tissue sarcomas. SUMMARY BACKGROUND DATA: Numerous studies on extremity soft tissue sarcomas have consistently shown that presentation with locally recurrent disease is associated with the development of subsequent local recurrences and that large tumor size and high histologic grade are significant factors associated with decreased survival. However, the effect of local recurrence on patient survival remains unclear. METHODS: From 1975 to 1997, 753 patients with intermediate- to high-grade extremity soft tissue sarcomas were treated at UCLA. Treatment outcomes and patient characteristics were analyzed to identify factors associated with both local recurrence and survival. RESULTS: Patients with locally recurrent disease were at a significantly increased risk of developing a subsequent local recurrence. Local recurrence was a morbid event requiring amputation in 38% of the cases. The development of a local recurrence was the most significant factor associated with decreased survival. Once a patient developed a local recurrence, he or she was about three times more likely to die of disease compared to similar patients who had not developed a local recurrence. CONCLUSIONS: Local recurrence in patients with intermediate- to high-grade extremity soft tissue sarcomas is associated with the development of subsequent local recurrences, a morbid event decreasing functional outcomes and the most significant factor associated with decreased survival. Although 85% to 90% of patients with high-grade extremity soft tissue sarcomas are treatable with a limb salvage approach, patients who develop a local recurrence need aggressive treatment and should be considered for trials of adjuvant systemic therapy. PMID- 12560781 TI - Application of staging systems for differentiated thyroid carcinoma in an endemic goiter region with iodine substitution. AB - OBJECTIVE: To evaluate and compare staging systems for differentiated thyroid carcinoma and predicted outcome in an endemic goiter region with iodine substitution and to examine the risk profile of differentiated thyroid carcinoma and compare it against nongoiter regions. SUMMARY BACKGROUND DATA: Differentiated (papillary or follicular) thyroid carcinoma has a favorable prognostic outcome. In numerous studies prognostic factors have been identified and staging systems created, particularly in Anglo-American centers (nonendemic goiter regions), to evaluate individual prognostic outcome. METHODS: In a retrospective study, the authors assessed 440 patients with differentiated thyroid carcinoma (papillary, n = 293; follicular, n = 147) and a long-term follow-up of median 10.6 years to determine the predictive accuracy of nine staging systems applicable to the study population; the systems were compared by calculating the proportion of variation explained. RESULTS: With regard to cause-specific mortality, the difference between the respective stages and/or risk groups was highly significant for every staging system. By means of calculating the proportion of variation explained, MACIS scoring supplied the most reliable prognostic information for differentiated thyroid carcinoma (relative importance 16.93%). EORTC and UICC/AJCC systems had a relative importance of 16.34% and 13.96%, respectively, also a high level of accuracy; this implies that they are superior to the other six staging systems. If we separate papillary and follicular carcinoma, for the former the MACIS score with a relative importance of 15.05% is clearly superior to the other staging systems, whereas for the latter the EORTC score and the UICC/AJCC staging system, with relative importance of 17.04% and 16.58%, respectively, yield the best prognostic information. CONCLUSIONS: By applying staging systems in an endemic goiter region with iodine substitution, the best prognostic information for papillary thyroid carcinoma has been achieved with the MACIS score, while for follicular thyroid carcinoma the EORTC score and the UICC/AJCC system have the best prognostic accuracy. Because of the individual factors, which are easy to obtain and generally available (age, T, N, M classification), the uncomplicated handling, and the widespread use and the good predictive accuracy, the UICC/AJCC classification is the staging system of choice for comparing published results. PMID- 12560782 TI - Ertapenem versus piperacillin/tazobactam in the treatment of complicated intraabdominal infections: results of a double-blind, randomized comparative phase III trial. AB - OBJECTIVE: To examine the clinical efficacy and safety of ertapenem, a novel beta lactam agent with wide activity against common pathogens encountered in intraabdominal infection. SUMMARY BACKGROUND DATA: Ertapenem has a pharmacokinetic profile and antimicrobial spectrum that support the potential for use as a once-a-day agent for the treatment of common mixed aerobic and anaerobic infections. METHODS This prospective, randomized, controlled, and double-blind trial was conducted to compare the safety and efficacy of ertapenem with piperacillin/tazobactam as therapy following adequate surgical management of complicated intraabdominal infections. RESULTS: Six hundred thirty-three patients were included in the modified intent-to-treat population, with 396 meeting all criteria for the evaluable population. Patients with a wide range of infections were enrolled; perforated or abscessed appendicitis was most common (approximately 60% in microbiologically evaluable population). A prospective, expert panel review was conducted to assess the adequacy of surgical source control in patients who were failures as a component of evaluability. For the modified intent-to-treat groups, 245 of 311 patients treated with ertapenem (79.3%) were cured, as were 232 of 304 (76.2) treated with piperacillin/tazobactam. One hundred seventy-six of 203 microbiologically evaluable patients treated with ertapenem (86.7%) were cured, as were 157 of the 193 (81.2%) treated with piperacillin/tazobactam. CONCLUSIONS: In this study, the efficacy of ertapenem 1 g once a day was equivalent to piperacillin/tazobactam 3.375 g every 6 hours in the treatment of a range of intraabdominal infections. Ertapenem was generally well tolerated and had a similar safety and tolerability profile to piperacillin/tazobactam. A formal process for review of adequacy of source control was found to be of benefit. The results of this trial suggest that ertapenem may be a useful option that could eliminate the need for combination and/or multidosed antibiotic regimens for the empiric treatment of intraabdominal infections. PMID- 12560783 TI - Protective effects of medium-chain triglycerides on the liver and gut in rats administered endotoxin. AB - OBJECTIVE: To determine if medium-chain triglycerides (MCTs) prevent organ injuries and mortality in rats administered endotoxin and to investigate effects of MCT on the gut. SUMMARY BACKGROUND DATA: Since dietary MCTs prevent alcohol induced liver injury by inhibiting activation of Kupffer cells in the enteral feeding model, the authors hypothesized that MCT could prevent deleterious conditions in endotoxemia. METHODS: After a preliminary experiment determined the optimal dose of MCT, rats were given MCT (5 g/kg per day) or the same dose of corn oil by gavage daily for 1 week. Then, lipopolysaccharide (LPS) was administered intravenously and survival was assessed for the next 24 hours. For analysis of mechanisms, rats were killed 9 hours after LPS injection and serum and liver sections were collected. To investigate effects of MCT on the gut, pathologic change, permeability, and microflora were assessed. Kupffer cells isolated by collagenase digestion and differential centrifugation were used for endotoxin receptor CD14 immunoblotting, phagocytic index, and TNF-alpha production assay. RESULTS: All rats given corn oil died after LPS administration; however, this mortality was prevented by MCT in a dose-dependent manner. Rats given corn oil showed liver injury after LPS administration. In contrast, MCT prevented this pathologic change nearly completely. MCT blunted CD14 expression on the Kupffer cells and TNF-alpha production by isolated Kupffer cells; however, there were no differences in phagocytic index between the two groups. The length of the intestinal epithelium was increased in the MCT group compared to the corn oil group. Further, after LPS administration, increases in gut permeability and injury were prevented by MCT. Importantly, MCT also prevented hepatic energy charge and gut injuries in this condition. CONCLUSIONS: Enteral feeding using MCT could be a practical way of protecting the liver and intestine during endotoxemia. PMID- 12560784 TI - Graft injury in relation to graft size in right lobe live donor liver transplantation: a study of hepatic sinusoidal injury in correlation with portal hemodynamics and intragraft gene expression. AB - OBJECTIVE: To investigate the degree and mechanism of hepatic sinusoidal injury in different graft sizes in right lobe live donor liver transplantation (LDLT). SUMMARY BACKGROUND DATA: Liver grafts from living donors are likely to be small for-size for adult recipients. Graft injury after reperfusion is common, but the mechanism and degree of injury remain unclear. The hepatic sinusoidal injury in different graft sizes and its relationship with portal hemodynamics and intragraft gene response at the early phase after reperfusion have not been studied in right lobe LDLT. METHODS: From May 2000 to November 2001, 40 adults receiving right lobe LDLT had portal pressure measured continuously before and after reperfusion. Liver biopsies were taken before and after reperfusion for detection of vasoregulatory genes (endothelin-1 and endothelial nitric oxide synthase) and heat shock genes (heat shock protein 70 and heme oxygenase-1), and electron microscope examination. Blood samples from the portal vein and suprahepatic inferior vena cava were taken for the measurement of plasma nitric oxide level. RESULTS: The recipients were grouped according to the ratio of graft weight to estimated standard liver weight: group 1 (n = 10), less than 40%; group 2 (n = 21), 40% to 60%; and group 3 (n = 9), more than 60%. The portal pressures recorded after reperfusion in group 1 were significantly higher within 30 minutes of reperfusion than those in groups 2 and 3. After reperfusion, the intragraft endothelin-1 mRNA level in group 1 increased by 161% of the basal level but decreased by 31.5% and 62% of the basal level in groups 2 and 3, respectively. The intragraft mRNA level of heme oxygenase-1 in groups 1 and 2 decreased by 75.5% and 25.3% of the basal level respectively but increased by 41% of basal level in group 3. The intragraft protein level of heat shock protein 70 decreased by 50 ng/mL after reperfusion in group 1 but increased by 12.4 ng/mL and 0.6 ng/mL in groups 2 and 3, respectively. The portal vein plasma nitric oxide level decreased more significantly after reperfusion in group 1 than in group 2. Electron microscope examination of liver biopsies in group 1 showed tremendous mitochondrial swelling as well as irregular large gaps between the sinusoidal lining cells. There were two hospital deaths in group 1 and none in the other two groups. CONCLUSIONS: Patients implanted with grafts less than 40% of standard liver weight suffered from transient portal hypertension early after reperfusion. The phenomenon was accompanied by intragraft upregulation of endothelin-1 and ultrastructural evidence of sinusoidal damage. The transient portal hypertension after reperfusion, subsequent endothelin-1 overexpression, and plasma nitric oxide level reduction, together with downregulation of heme oxygenase-1 and heat shock protein 70, may account for the small-for-size graft injury. PMID- 12560785 TI - Heart xenograft survival with chimeric pig donors and modest immune suppression. AB - OBJECTIVE: To assess the use of donor pigs with cellular chimerism for prevention of acute rejection with modest immune suppression. The clinical use of pig organ xenografts is currently precluded by severe acute rejection, which resists standard immune suppression. SUMMARY BACKGROUND DATA: For long-term survival of pig organ xenografts, immune suppression significantly greater than used with allografts would currently be necessary, leaving the recipient immune deficient and at increased risk for infections. Induction of immune tolerance and tissue accommodation could enhance xenograft survival but would lead to complications and frequent graft failure. Induction of cellular chimerism within the donor pigs, however, could accomplish these goals before transplantation, significantly reducing the risk. METHODS: Marrow cells from sheep were infused into fetal pigs. Heart xenografts from chimeric or nonchimeric pigs were transplanted heterotopically into recipient sheep, simultaneous with infusion of splenocytes. Posttransplant suppression consisted of cyclosporine and tapered corticosteroids, comparable with allotransplants. RESULTS: All of the control grafts (n = 12) were rejected by acute vascular rejection in 4 to 8 days. In contrast, only one episode of vascular rejection was observed in the experimental group (n = 13). Four experimental recipients had an episode of moderate diffuse cellular rejection (grade 3) and one had moderate focal cellular rejection (grade 2). Each episode responded to pulse steroids. Seven grafts showed no significant rejection. There was little evidence of immune deficiency, infection, or toxicity. CONCLUSIONS: Acute vascular rejection was prevented in a large animal model without the need for severe immune suppression. PMID- 12560786 TI - Gallstone disease in heart transplant recipients. AB - OBJECTIVE: To review the outcome of cholecystectomy after heart transplant. SUMMARY BACKGROUND DATA: The optimal timing for gallbladder surgery in heart transplant patients is controversial. METHODS: Between April 1985 and October 2000, 518 cardiac transplants were performed at Ochsner Foundation Hospital. Data gathered included ultrasound reports, cholecystectomy operative reports, gallbladder pathologic reports, complications, and deaths. RESULTS: Charts were available for 509 patients (98%), 68 (13%) of whom underwent cholecystectomy before transplantation. Of the 509, 53 (10%) had serial ultrasound examinations and 29 of the 53 (55%) developed gallstones. After transplant, 47 (9%) underwent cholecystectomy. Five cholecystectomies were performed during the immediate postoperative course. Two patients who underwent cholecystectomy had acalculous cholecystitis; one was incidental. Four patients died (one with rejection and three with sepsis). After discharge, 42 cholecystectomies were performed: 16 for biliary colic (no deaths, three patients with complications), 19 for acute cholecystitis (one death, nine patients with complications), 5 for biliary pancreatitis (1 death, 1 patient with complications), and 2 others. CONCLUSIONS: The risk of morbidity and mortality from gallstone disease is high in cardiac transplant patients, particularly immediately posttransplant. Posttransplant patients require annual ultrasound examinations to detect the onset of gallstone disease, and this risk is higher than in the general population. Gallstones alone are an indication for cholecystectomy in the cardiac transplant patient. Pretransplant cholecystectomy should be considered in clinically stable patients with gallstones. PMID- 12560787 TI - Influence of bilateral skeletonized harvesting on occurrence of deep sternal wound infection in 1,000 consecutive patients undergoing bilateral internal thoracic artery grafting. AB - OBJECTIVE: To evaluate the risk of deep sternal infection in a large patient cohort following bilateral internal thoracic artery (BITA) grafting using skeletonized BITA dissection. SUMMARY BACKGROUND DATA: Complete myocardial revascularization using BITAs improves long-term survival and lowers the rate of repeat operations. Harvesting of ITAs as skeletonized vessels preserves sternal collateral blood supply, thus enabling rapid sternal healing with less risk of deep sternal infection. METHODS: One thousand consecutive patients (763 men, 340 patients >70 years old, 304 diabetics) underwent skeletonized BITA grafting from April 1996 to July 1999. RESULTS: The 30-day mortality rate was 3.4%. There were 10 perioperative infarcts, 16 strokes, and 22 deep sternal infections. There was an increased risk of deep sternal infection in repeat coronary artery bypass grafting (CABG) operations (15%), chronic obstructive pulmonary disease (COPD) (6.2%), congestive heart failure (4.7%), left ventricular dysfunction (ejection fraction < 35%, 4.5%), and longer aortic cross-clamping time. After adjustment for other demographic, clinical, and surgical predictors, the only independent predictors of deep sternal infection were repeat operations, COPD, and duration of aortic cross-clamping. No patients in the reoperation subgroup died, but three of six COPD patients with deep sternal infection died, and COPD was an independent predictor of overall (early + late) mortality. CONCLUSIONS: Skeletonized BITA grafting carries an acceptable risk of deep sternal infection but is not recommended for repeat CABG or for patients with COPD. PMID- 12560788 TI - Clinical and hematologic benefits of partial splenectomy for congenital hemolytic anemias in children. AB - OBJECTIVE: To assess the role of partial splenectomy for symptomatic children with various congenital hemolytic anemias. SUMMARY BACKGROUND DATA: The use of total splenectomy for symptomatic children with congenital hemolytic anemias is restricted by concern of postsplenectomy sepsis. A partial splenectomy is an alternative procedure, although its utility remains incompletely defined. METHODS: This longitudinal cohort study followed 25 symptomatic children with various congenital anemias who underwent partial splenectomy. Sixteen children had hereditary spherocytosis (HS), and nine children had other erythrocyte disorders. Outcome measures were clinical and laboratory hemolysis, splenic phagocytic and immune function, and splenic regrowth as measured by ultrasonography. Discrete parameters were compared using the Student test. RESULTS: Partial splenectomy was successful in all 25 children, with minimal morbidity. Follow-up ranged from 7 months to 6 years (mean 2.3 +/- 1.5 years). Following surgery, children with HS had increased hemoglobin values, decreased reticulocyte and bilirubin levels, and preserved splenic function. Most children without HS had decreased symptoms of hypersplenism and splenic sequestration. Over time, variable rates of splenic regrowth were noted, although regrowth did not necessarily correlate with recurrent hemolysis. CONCLUSIONS: In children with hereditary spherocytosis, a partial splenectomy appears to control hemolysis while retaining splenic function. In children with other congenital hemolytic anemias, a partial splenectomy appears to control symptoms of hypersplenism and splenic sequestration. PMID- 12560789 TI - Detection of response to neoadjuvant therapy of esophageal squamous cell carcinoma by positron emission tomography (PET). PMID- 12560791 TI - Using a model for structured reflection on palliative care nursing: exploring the challenges raised. AB - Reflection on palliative care nursing practice has to encompass every dimension of the holistic care we offer. In this article a possible structured model for reflection on palliative care nursing is discussed and used to reflect on the care given during a community Macmillan nurse visit. The reflection revealed a number of challenges. These included the possible conflict between patient autonomy and quality of life for carers; the role of intuition in palliative nursing and spiritual care. The article concludes that reflection has a role to play in exploring the innate tensions in palliative care, voicing the often silent intuitive knowledge and promoting expertise in spiritual care. PMID- 12560792 TI - Human research ethics committees: issues in palliative care research. AB - Palliative care research is fraught with many difficulties. There are challenges associated with conducting research with vulnerable patients and families, difficulties with obtaining informed consent, and methodological complexities. Thoughtful construction of research protocols may overcome many of these problems. However, researchers may be powerless to overcome the discomfort of members of human research ethics committees (HRECs) who disallow access to palliative care patients and families. The notion of conducting research with this group is often perceived as abhorrent by those who do not practise in palliative care. This is because of a persistent idea that dying people and their families are so burdened by the dying process and so vulnerable to exploitation that they should not be approached to be involved in research. This over protectiveness regarding palliative care research often distorts the proper gate keeping role of HRECs and health-care professionals. This article draws on the authors' experiences of presenting applications to HRECs over the last 20 years. It explores the responsibilities of HRECs, the responsibilities of palliative care researchers and the rights of patients and families. HRECs and health professionals who endeavour to undertake palliative care research are encouraged to reflect and re-examine the role of ethics committees. PMID- 12560793 TI - A reflection on Cicely Saunders' views on a good death through the philosophy of Charles Taylor. AB - In her writings, Cicely Saunders has developed a total framework of caring for seriously ill people. Saunders' reaction to what she has experienced as bad contemporary post-war health care is to be found in her hospice philosophy. In this article this philosophy is analysed through the writings of Charles Taylor. Taylor is a Canadian philosopher who has delivered important views on what it means to live a 'good' and 'authentic' life in a modern society. By focusing on Saunders' writings from Taylor's perspective it becomes apparent that Saunders has created a very modern theory. The author has found that different aspects of a good and authentic life are important values in Saunders' good and authentic death. These findings are followed by one question: is Saunders' hospice philosophy too focused on values such as individuality, control and autonomy, giving too little attention to fragility and dependency? PMID- 12560794 TI - Communication in the development of culturally competent palliative care services in the UK: a case study. AB - Research suggests that many minority ethnic patients who receive palliative care in the UK are satisfied with the service they are given. However, various studies have revealed that minority ethnic groups' experiences of care are far from perfect. The most significant problem for these patients centres on communication. This article presents some results from an exploratory study, commissioned by Luton Health Action Zone, to explore the role of communication in delivering effective palliative care services to South Asians living in Luton. Overall, it was found that the services provided are, in most cases, valued and seen as being effective. However, as the service providers who were interviewed readily recognized, there were areas where improvements could be made. The main issues were found to be the need to inform South Asian populations of the availability of palliative care services and the need to improve communication between patients and service providers. This article describes the communication problems that service providers and users face. It also identifies possible policy improvements aimed at developing the 'cultural competency' of services. PMID- 12560795 TI - Learning together: use of simulated patients with nursing and medical students for breaking bad news. AB - This article reports a small research study on the use of simulated patients for teaching medical and nursing students how to break bad news. A total of 34 nursing and medical students undertook the activity in mixed groups. Data obtained from free response questionnaires and a focus group were analysed and independently coded to identify recurrent themes across the data and the two student groups. Students reported that simulated patients gave instant feedback about their performance which they found useful. Students felt that working in doctor-nurse pairs helped them formulate strategies for imparting the bad news and that this made them feel better prepared and less isolated. The article outlines why the use of simulated patients may be valuable and suggests that this form of teaching could be used in other types of communications training. The use of simulated patients appears to have the potential to enhance patient practitioner and practitioner-practitioner interactions. PMID- 12560796 TI - A commentary on 'An integrated care pathway for the last two days of life'. PMID- 12560797 TI - The importance of social-historical context. PMID- 12560798 TI - An integrated care pathway for the last two days of life: Wales-wide benchmarking in palliative care. AB - Functional benchmarking assesses performance and practice across a broad range of settings and carries the potential to effect change in practice. An integrated care pathway (ICP) can assist in the benchmarking process, defining desired outcomes for specific patient groups over a designated time frame. Any variations to the agreed course of care are documented using the 'variance sheet'. This article describes the Wales-wide implementation of an ICP for the last two days of life. The project has enabled an ongoing centralized collection and analysis of variance sheets, which reflect the care of the dying patient in four different care settings crossing the voluntary and statutory sectors. Initial analysis of the first 500 variance sheets to be generated by the ICP for the last two days of life indicates that the management of pain, agitation, excess respiratory secretions and mouth care may be problematic. The same problems were experienced across acute, hospice, specialist inpatient units and community care. Closing the audit cycle involves incorporating the information from the variance analysis into clinical practice. PMID- 12560799 TI - Meeting complex needs: an analysis of Macmillan nurses' work with patients. AB - Macmillan nurses, working as Clinical Nurse Specialists (CNS), are in many ways the public face of specialist palliative care in the UK. However, the way in which they manage their clinical workload is not well understood. Questions about this aspect of their work have emerged from a large-scale evaluation of Macmillan nursing in 12 sites in the Trent and Thames regions of the UK, funded by Macmillan Cancer Relief. This article focuses on how Macmillan nurses enact their patient care role, drawing on caseload data, diaries and semi-structured interviews from the wider evaluation. It concentrates on the following issues: characteristics of referred patients; nature of interventions given; division of time between different aspects of the CNS role; nurses' perceptions of their patient-care role. Findings suggest that activities associated with direct patient care account for the majority of nurses' time, with little time spent in any other individual aspect of the CNS role. The delivery of care to older patients at the end of life is a major part of Macmillan nurses' clinical role. Issues relating to the educational and support needs of Macmillan nurses are raised. PMID- 12560800 TI - Living on the outside looking in: a theory of living with advanced breast cancer. AB - Much of the literature on women's experiences of breast cancer relates to diagnosis and initial treatment. The aim of this research was to develop a theory to explain the meaning and experience of living with advanced breast cancer. A grounded theory approach was used and ten women under the care of one hospice explained their experiences during semi-structured interviews. The findings suggest that there are significant issues and difficulties facing women with advanced breast cancer, culminating in feelings of isolation and the experience of living with the fear of advancing disease. A theory, living on the outside looking in, was developed to explain the experiences of the participants. This integrates the following five categories: reconciling a different me; time bomb; media effect; professional waning; rescuers. The category 'reconciling a different me' was identified as the core variable and had an explicatory relationship to the other four categories. The findings raise issues regarding appropriate support. PMID- 12560801 TI - Use of quality-of-life scores in care planning in a hospice setting: the theme of revelation. AB - This article follows an earlier one in which measurement of quality of life (QOL) was used in developing patient-care plans and to identify differences in QOL assessment between nurses and patients as an aid to reflective practice. The present article discusses how the theme of revelation emerged from the insights of those in the study. The process of measuring, and subsequent use of, QOL scores increased understanding between nurse and patients. Some nurses were surprised at how different their perception of a patient's QOL was from that of the patient. Completing QOL questionnaires also gave some patients new insights. How such revelations might be used to improve QOL are discussed. PMID- 12560802 TI - Palliative care for Muslims and issues after death. AB - In this, our second paper looking at issues in providing palliative care to Muslims in the UK, we focus on death customs and bereavement. We cover issues relating to the immediate handling of the body after death, washing the deceased and reasons for early burial. Muslim views on post-mortems and organ transplantation are explored. In addition to sharing information on practices common among Muslim communities, we also provide practical advice on the day-to day issues that may arise when caring for a 'recently deceased' Muslim patient and his or her relatives. PMID- 12560803 TI - A commentary on 'Joe's story: reflections on a difficult interaction between a nurse and a patient's wife'. PMID- 12560804 TI - Crash proof kids? An overview of current motor vehicle child occupant safety strategies. PMID- 12560806 TI - The chicken as a model for large-scale analysis of vertebrate gene function. PMID- 12560807 TI - Signatures of natural selection in the human genome. PMID- 12560808 TI - Caenorhabditis elegans operons: form and function. PMID- 12560809 TI - Ancient horizontal gene transfer. PMID- 12560810 TI - Pattern formation: old models out on a limb. PMID- 12560811 TI - Studying complex biological systems using multifactorial perturbation. PMID- 12560812 TI - Of eponyms, acronyms and...orthonyms. PMID- 12560813 TI - From Cambridge to Srebrenica. PMID- 12560814 TI - Organizing mRNA export. PMID- 12560815 TI - Surprise ending. PMID- 12560816 TI - Dissecting dyskeratosis. PMID- 12560818 TI - Sciona and genetic testing. PMID- 12560820 TI - Generating copy(s). PMID- 12560821 TI - Ambitious French biotech plan faces EU hurdle. PMID- 12560822 TI - Cash crunch quells antibody litigation. PMID- 12560823 TI - Lymphoma market turf war imminent, pending Bexxar approval. PMID- 12560824 TI - US policy on data disclosure debated. PMID- 12560825 TI - India set to embrace GM rice. PMID- 12560826 TI - US policy may encourage counterfeit drugs. PMID- 12560827 TI - The changing norms of the life sciences. PMID- 12560828 TI - The price of conserving agricultural biodiversity. PMID- 12560829 TI - Let the chips fall where they may. PMID- 12560830 TI - Outside expertise. PMID- 12560831 TI - RNAi targeting of DNA virus in plants. PMID- 12560837 TI - Rewiring yeast for drug synthesis. PMID- 12560838 TI - Vitamin C booster. PMID- 12560839 TI - Pseudomonas putida--a versatile biocatalyst. PMID- 12560840 TI - Designer milk from transgenic clones. PMID- 12560844 TI - Workplace programs to protect trade secrets. PMID- 12560848 TI - Transferable skills for a competitive edge. PMID- 12560849 TI - Should the CAG advocate for resources with government and governmental agencies? PMID- 12560851 TI - Hepatology and the Canadian gastroenterologist: interest, attitudes and patterns of practice: results of a national survey from the Canadian Association of Gastroenterology. AB - BACKGROUND: Hepatology has emerged as a subspecialty distinct from gastroenterology. Despite this, there is no formal certification examination or accredited training program, and training remains combined with gastroenterology. AIM: To determine attitudes, perceptions and patterns of practice with respect to liver disease among Canadian gastroenterologists. METHODS: A survey questionnaire was distributed to clinician gastroenterologists who are members of the Canadian Association of Gastroenterology. The responses of subgroups of respondents were compared by univariate and multivariate statistical techniques. RESULTS: Hepatologists constituted 20 of 201 respondents, the rest identifying themselves as gastroenterologists. Among gastroenterologists, liver disease constituted 10% of in- and out-patient practice. Despite this, 85% of gastroenterologists maintain an interest in hepatology, 49% perform liver biopsies, 60% treat hepatitis C, and 54% treat hepatitis B. In all of these areas, university-based gastroenterologists were consistently less likely than community-based gastroenterologists to maintain an interest and practice in hepatology, a finding that remained statistically significant on multivariate analysis. With regard to hepatology training, 90% of hepatologists and 94% of gastroenterologists felt that hepatology training should remain combined with gastroenterology, although 55% of hepatologists felt that current training was adequate compared with 79% of gastroenterologists, who were satisfied with the status quo. CONCLUSIONS: Hepatology remains relevant and important to Canadian gastroenterologists, especially those who have community-based practices. Canadian gastroenterologists and hepatologists are not in favour of separating hepatology training from existing gastroenterology training programs, although hepatologists feel that the current level of training is suboptimal. PMID- 12560852 TI - Role of Na+/H+ exchanger isoform-1 in human inflammatory bowel disease. AB - BACKGROUND: Na+/H+ exchanger (NHE) is responsible for a net uptake of sodium chloride and water from the gastrointestinal tract and maintains electrolyte and water homeostasis. However, its status in human inflammatory bowel disease such as ulcerative colitis (UC) and Crohn's disease (CD) remains poorly understood. OBJECTIVES: To investigate the role of NHE-1 isoform in human CD and UC. METHODS: Expression of NHE-1 protein and messenger ribonucleic acid and sodium pump activity were examined in the colonic biopsy samples taken from UC (n=11) and CD (n=13) patients using enhanced chemiluminescence-Western blot analysis, reverse transcription polymerase chain reaction and spectrophotometry. Subjects presenting with abdominal pain and endoscopically normal colon served as normal controls (n=11). Myeloperoxidase (MPO) activity and histology were performed to confirm tissue inflammation. RESULTS: MPO activity increased significantly (P<0.05) in both UC and CD patients compared with the normal controls. Parallel to MPO activity profile, there was also a significantly higher infiltration of inflammatory cells in both cases. P-nitrophenylphosphatase activity, a marker of the sodium pump, remained unchanged in CD but increased significantly (P<0.05) in UC compared with the normal controls. On the contrary, the level of NHE-1 protein and messenger ribonucleic acid was significantly decreased (P<0.05) in both cases, whereas the internal control, a-actin remained unaltered. CONCLUSIONS: These findings demonstrate a transcriptionally regulated suppression of NHE-1 in both UC and CD. This NHE-1 suppression may reduce an uptake of sodium chloride and water from the inflamed colonic lumen and thus contribute to diarrhea and neuromuscular alterations in these conditions. PMID- 12560853 TI - Nonalcoholic fatty liver disease in patients investigated for elevated liver enzymes. AB - Nonalcoholic fatty liver disease (NAFLD) is a common diagnosis among patients referred to gastroenterology and hepatology clinics for the evaluation of elevated liver enzymes. The diagnosis of NAFLD is supported by blood work to exclude other liver diseases, and by ultrasound evidence of fat in the liver in patients without a significant history of alcohol intake. The gold standard, however, is a liver biopsy to show the typical histological features of NAFLD, which are almost identical to those of alcohol-induced liver damage and can range from mild steatosis to cirrhosis. A variety of retrospective series have linked NAFLD to obesity, diabetes, hyperlipidemia, total parenteral nutrition, jejunoileal bypass surgery and certain medications. A subset of patients with NAFLD that had an initial presentation of elevated liver enzymes was studied. Two hundred and two patients were reviewed, of whom 49 met the inclusion criteria including a liver biopsy. Patients were excluded if insufficient data were available, if the patients had a significant history of ethanol intake or if they had other coexisting liver disease. These patients were seen between 1996 and 2000 in gastroenterology and hepatology clinics in two community hospitals and one regional liver transplant centre in Edmonton, Alberta. NAFLD was associated with a spectrum of changes in the liver ranging from mild steatosis to more significant steatosis with inflammation and fibrosis. Cases of NAFLD with steatosis and mixed inflammatory infiltration but lacking ballooning degeneration or fibrosis were prevalent in young (20 to 40 years of age) patients with no other significant medical history except for obesity. NAFLD with biopsies showing significant fibrosis and ballooning cell degeneration was associated with obesity, diabetes and older age. It was concluded that, in this predominantly outpatient setting, age over 40 years and diabetes at any age are risk factors for both nonalcoholic steatohepatitis and nonalcoholic steatohepatitis with cirrhosis. It is therefore recommended that patients with raised liver enzymes and suspected NAFLD be targeted for liver biopsy in their evaluation. PMID- 12560854 TI - Appendiceal carcinoids in Crohn's disease. AB - Earlier investigations demonstrate an increased risk for colon cancer in Crohn's disease. For other intestinal neoplasms, such as carcinoids, studies are limited. In Crohn's disease, repeated endoscopic and imaging studies along with intestinal resections may facilitate clinical recognition of neoplastic diseases, including appendiceal neoplasms. To date, however, only sporadic cases of appendiceal carcinoids have been described in Crohn's disease. In the present study, in a single clinician database of 1000 Crohn's disease patients, three of the 441 patients who had undergone intestinal resection had appendiceal carcinoids, all of which were pathologically confirmed. All were observed in female patients and were not suspected before surgical treatment. In one case, even though management was not altered, the tumour had already invaded serosal fat indicating a potential for more advanced disease. In this series, a carcinoid tumour was found in a resection specimen during a later clinical case review and another was a microcarcinoid, implying that these tumours may be overlooked in Crohn's disease. The percentage detected in the entire database (0.3%) exceeds the reported rates of detection of appendiceal carcinoids after removal of the appendix for appendicitis, as well as the rate of detection of appendiceal carcinoids in autopsy studies. This percentage would be higher if only those having an intestinal resection were considered (0.68%). Additional studies are needed to further define this risk of appendiceal carcinoids in Crohn's disease. PMID- 12560855 TI - Motion--genetic testing is useful in the diagnosis of nonhereditary pancreatic conditions: arguments for the motion. AB - Mutations of three major genes are associated with an increased risk of acute and chronic pancreatitis: the cationic trypsinogen (PRSS1) gene, the cystic fibrosis transmembrane conductance regulator (CFTR) gene, and the pancreatic secretory trypsin inhibitor (PSTI) or serine protease inhibitor, Kazal type 1 (SPINK1) gene. Some autosomal dominant forms of hereditary pancreatitis are associated with mutations of the PRSS1 gene, which can be readily identified by genetic testing. Mutations of the CFTR gene can lead either to cystic fibrosis or to idiopathic chronic pancreatitis, and to a variety of cystic fibrosis-associated disorders, including congenital bilateral absence of the vas deferens and sinusitis. These mutations, as with those of the SPINK1 (or PSTI) gene, are prevalent in North America; thus, the presence of such a mutation in an asymptomatic person does not confer a high risk of developing pancreatitis. Combinations of mutations of the PRSS1 and SPINK1 genes lead to more severe disease, as indicated by an earlier onset of symptoms, which suggests that SPINK1 is a disease modifier. The major fear expressed by potential candidates for genetic testing is that the results could lead to insurance discrimination. Studies of the positive predictive value of genetic tests are hampered by recruitment bias and lack of knowledge of family history of pancreatitis. Genetic testing is most useful for persons for whom family members have already been found to exhibit a particular pancreatitis-associated mutation. In the future, increased knowledge of the myriad genetic causes of pancreatitis, as well as advances in the diagnosis and treatment of early chronic pancreatitis, should enhance the utility of genetic testing. PMID- 12560856 TI - Motion--genetic testing is useful in the diagnosis of nonhereditary pancreatic conditions: arguments against the motion. AB - Mutations of two genes, the cystic fibrosis transmembrane conductance regulator gene (CFTR) and the pancreatic secretory trypsin inhibitor gene (PSTI), are associated with an increase in the risk of idiopathic chronic pancreatitis. Persons who have mutations of both CFTR alleles (one severely and one mildly affected) are especially susceptible to this disease. Because these compound heterozygotes have sufficient residual CFTR function, they do not develop cystic fibrosis lung disease. One PSTI mutation, N34S, independently increases the risk of pancreatitis. Thus, the risk of pancreatitis is greatest among individuals who are CFTR compound heterozygotes and who also have the PSTI mutation. Nonetheless, most people with CFTR and PSTI mutations do not develop pancreatitis. This fact indicates that environmental influences and gene-gene interactions also affect pancreatitis risk. Although CFTR and PSTI genetic testing can identify persons at an increased risk of pancreatitis, there are several reasons why the routine screening of individuals with nonhereditary pancreatitis is not recommended at this time: most disease-associated mutations are not detected by readily available techniques, genetic counselling guidelines do not exist, most patients with mutations do not develop pancreatitis and the results of testing do not affect the clinical management of pancreatitis. PMID- 12560857 TI - Motion--pancreatic endoscopy is useful for the pain of chronic pancreatitis: arguments for the motion. AB - Pain is the dominant clinical problem in patients with chronic pancreatitis. It can be due to pseudocysts, as well as strictures and stones in the pancreatic ducts. Most experts agree that obstruction could cause increased pressure within the main pancreatic duct or its branches, resulting in pain. Endoscopic therapy aims to alleviate pain by reducing the pressure within the ductal system and draining pseudocysts. Approaches vary according to the specific nature of the problem, and include transgastric, transduodenal and transpapillary stenting and drainage. Additional techniques for the removal of stones from the pancreatic duct include extracorporeal shockwave lithotripsy. Success rates for stone extraction and stenting of strictures are high in specialized centres that employ experienced endoscopists, but pain often recurs during long term follow-up. Complications include pancreatitis, bleeding, infection and perforation. In the case of pancreatic pseudocysts, percutaneous or even surgical drainage should be considered if septae or large amounts of debris are present within the lesion. This article describes the techniques, indications and results of endoscopic therapy of pancreatic lesions. PMID- 12560858 TI - Motion--pancreatic endoscopy is useful for the pain of chronic pancreatitis: arguments against the motion. AB - Endoscopic therapy can be used to dilate strictures in the pancreatic duct, remove stones and drain pseudocysts. In addition, it provides an alternative to surgery for the management of pain in patients with chronic pancreatitis. Pain is a difficult problem in these patients, especially if substance abuse is present, and its medical management is generally unsatisfactory. The concept that pancreatic pain is related to increased pressure in the main pancreatic duct is unproven, and is not supported by the results of surgical intervention. Although pancreatic stenting is often technically successful at achieving drainage of the pancreatic duct and relieving pain over the short term, pain usually recurs with time, complications are frequent, and repeated stent changes are usually necessary. Pancreatic pseudocysts can be drained endoscopically, using transpapillary, cystogastrostomy or cystoduodenostomy approaches, but success rates are less than 50% and bleeding is a major complication. Pseudocysts should not be drained unless they are symptomatic, causing complications or enlarging. There have been no published studies comparing endoscopic with surgical or radiological modalities. Endoscopic therapy of pancreatic disorders is a new and interesting technique, but initial promising results need to be confirmed in large, well-designed clinical trials. Such studies would need to enrol large numbers of patients, and involve measurement of technical success, pain severity and quality of life parameters. At present, endoscopic techniques must be considered experimental. PMID- 12560869 TI - Large variation in MRSA policies, procedures and prevalence in English intensive care units: a questionnaire analysis. AB - OBJECTIVE: Methicillin-resistant Staphylococcus aureus (MRSA) is a major problem in intensive care units in most countries. Despite recommendations for screening and isolation of patients with MRSA our perception has been that there is little uniformity in approach in ICUs besides adherence to basic infection control procedures. We thus sought to identify MRSA prevalence and the variation of infection control policy across intensive care units in England. DESIGN AND SETTING: Postal questionnaire with telephone follow-up in English intensive care units. MEASUREMENTS AND RESULTS: Responses were obtained from 217 (96%) ICUs. Marked variation in practice was noted in terms of patient screening, staff screening, infection control procedures, isolation or cohorting of colonised/infected patients, and ward discharge policy. Point prevalence data showed that 16.2% of ICU patients were known to be colonised or infected with MRSA. There was a regional bias, but no difference was noted between high and low prevalence regions in terms of unit demographics or infection control policies. CONCLUSIONS: This study highlights the lack of consistent policy across English ICUs regarding isolation, screening and discharge practices for MRSA. Prospective studies are urgently needed to determine best practice. PMID- 12560870 TI - Microalbuminuria does not reflect increased systemic capillary permeability in septic shock. AB - OBJECTIVE: To investigate the correlation between microalbuminuria and extravascular lung water in patients in septic shock who require mechanical ventilation for severe respiratory failure. DESIGN AND SETTING: Prospective, observational, clinical study in the 20-bed intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: 25 consecutive patients in septic shock and also in severe respiratory failure requiring mechanical ventilation. INTERVENTIONS: Hemodynamic parameters and extravascular lung water were determined by single arterial thermodilution. Together with each hemodynamic measurement the PaO(2)/FIO(2) ratio and urinary microalbumin to creatinine ratio (M:Cr) was measured. Serum C-reactive protein (CRP) and procalcitonin (PCT) levels were also determined daily. MEASUREMENTS AND RESULTS: The EVLW index was significantly higher than normal throughout the study. Microalbuminuria was in the normal range on entry and remained so for the rest of the study period. Serum PCT and CRP levels were significantly higher than normal at every assessment points. No significant correlation was found between M:Cr and either EVLW or PaO(2)/FIO(2). CONCLUSIONS: In this study patients in septic shock with significantly elevated EVLW had normal urinary M:Cr, and there was no correlation between M:Cr and EWLV, and PaO(2)/FIO(2). Therefore based on the current results routine measurements of microalbuminuria to determine endothelial permeability cannot be recommended in critically ill patients. PMID- 12560871 TI - Genotype and haplotype distributions of MTHFR677C>T and 1298A>C single nucleotide polymorphisms: a meta-analysis. AB - Common single nucleotide polymorphisms (SNPs; 677C>T and 1298A>C) in the methylenetetrahydrofolate reductase gene ( MTHFR) decrease the activity of the enzyme, leading to hyperhomocysteinemia, particularly in folate-deficient states. We calculate herein the haplotype frequencies of the MTHFR 677 and 1298 polymorphisms in pooled general populations derived from published data. We selected 16 articles that provided reliable data on combined MTHFR genotypes in general populations ( n = 5389). The combined data comprised the following totals for each genotype at nucleotide positions 677 and 1298: 838 CC/AA (i.e., 677CC/1298AA), 1225 CC/AC, 489 CC/CC, 1120 CT/AA, 1093 CT/AC, 8 CT/CC, 606 TT/AA, 10 TT/AC, and 0 TT/CC. The estimated haplotype frequencies, and the fractional contribution of each, were 677C/1298A, 0.37; 677C/1298C, 0.31; 677T/1298A, 0.32; and 677T/1298C, 0.0023 to 0.0034. Thus, a vast majority of 677T alleles and 1298C alleles are associated with 1298A alleles and 677C alleles, respectively. There may be an increased frequency of the very rare cis 677T/1298C haplotype in some parts of the United Kingdom and Canada, possibly due to a founder effect. Further studies on both SNPs are needed to determine their exact role in various clinical settings. PMID- 12560872 TI - A novel splice site mutation in neonatal carnitine palmitoyl transferase II deficiency. AB - Mitochondrial beta-oxidation of long-chain fatty acids requires the concerted action of three tightly integrated membrane-bound enzymes (carnitine palmitoyltransferase I and II and carnitine/acylcarnitine translocase) that transport them into mitochondria. Neonatal onset of carnitine palmitoyltransferase II (CPT II) deficiency is an autosomal recessive, often lethal disorder of this transport. We describe a novel splice-site mutation in the CPT II gene, found in a Moroccan family, of which four out of five children have died from the neonatal form of CPT II deficiency. Mutation detection studies at the mRNA level in the CPT II gene implied that the affected children were homozygous for the previously reported 534T insertion followed by a 25-bp deletion (encompassing bases 534-558). Studies of genomic DNA, however, revealed all patients to be compound heterozygous for this 534T ins/del 25 mutation, and for a new g-->a splice-site mutation in the splice-acceptor site of intron 2. Because of these findings, prenatal diagnosis was performed in chorionic villi of three new pregnancies. This did not reveal new compound heterozygous genotypes, and, after uneventful pregnancies, all children appeared to be healthy. The new mutation is the first splice-site mutation ever identified in CPT II deficiency. The fact that it was not discovered in the patient's cDNA makes this study another example of the incompleteness of mutation detection at the mRNA level in cases where a mutation leads to aberrant splicing or nonsense-mediated messenger decay. PMID- 12560873 TI - The interleukin 6-174G/C polymorphism is associated with indices of obesity in men. AB - Obesity represents an expansion of adipose tissue (AT) mass and is closely related to insulin resistance and cardiovascular disease. Several hormonal signals have been shown to originate from AT, one of them being interleukin 6 (IL6), for which one third of circulating levels is accounted for by AT. To study the impact of the IL6 -174G/C polymorphism on obesity-related phenotypes, we genotyped a cohort of 270 French-Canadian men from the greater Quebec City area selected to cover a wide range of body fatness values. The IL6 -174G allele was more commonly observed among lean subjects (body mass index <25 kg/m(2), chi(2) = 7.27, P = 0.007 or waist-line <100 cm, chi(2) = 6.63, P = 0.01). When men were subdivided according to insulin and glucose levels at 180 min following the glucose load, using 160 pmol/l and 4.6 mmol/l, respectively, as cutoff points, the -174G allele was more frequently observed in groups with low concentrations of either insulin or glucose, P = 0.03 and P = 0.01, respectively. When comparisons between genotype groups were performed, -174G/G homozygotes presented the lowest waist circumference ( P < 0.05). In summary, this study showed that, in men, the IL6 -174G/C polymorphism is associated with some indices of body composition and parameters of glucose and insulin homeostasis. PMID- 12560874 TI - DNA polymorphism and mutations in CPN1, including the genomic basis of carboxypeptidase N deficiency. AB - Carboxypeptidase N (EC 3.4.17.3) regulates the activity of peptides such as kinins and anaphylatoxins. Although deficiency of carboxypeptidase N (MIM 212070) produces a severe allergic syndrome, no human mutations have ever been described. Therefore, using archival genomic DNA from a subject with documented carboxypeptidase N deficiency, we sequenced CPN1 (MIM 603103), which encodes the catalytic subunit of carboxypeptidase N. In the genomic DNA of the proband, we discovered three CPN1 variants: (1) 385fsInsG, a frameshift mutation in exon 1 due to a single G insertion at nucleotide 385; (2) 746G>A single-nucleotide polymorphism (SNP), a missense mutation in exon 3 that predicted substitution of aspartic acid for the wild-type conserved glycine at amino acid 178 (G178D); and (3) IVS1 +6C>T, an SNP in intron 1. Among 128 normal Caucasians, the 385fsInsG mutation was absent and the G178D mutation had a frequency of 0.0078, suggesting that these were rare molecular events that likely contributed to the carboxypeptidase N deficiency phenotype. The frequency of the IVS1 +6C>T polymorphism was 0.051. The reagents described here provide tools for further study of association with clinical and biochemical phenotypes related to allergy and immunity. PMID- 12560875 TI - Catalog of 668 SNPs detected among 31 genes encoding potential drug targets on the cell surface. AB - We have been publishing a series of detailed maps of single-nucleotide polymorphisms (SNPs) detected within the genomic loci of 145 genes encoding drug metabolizing enzymes and transporters. As an addition to the maps reported earlier, we provide here high-density SNP maps of 31 genes encoding various receptors and adhesion molecules of medical importance. By examining a total of approximately 382 kb of genomic DNA encompassing these 31 genes, we identified 668 SNPs among 48 healthy Japanese individuals: 86 in 5' flanking regions, 27 in 5' untranslated regions, 45 in coding regions, 399 in introns, 47 in 3' untranslated regions, and 64 in 3' flanking regions. We also discovered 113 variations of other types. Of the 668 SNPs, 371 (55.5%) appeared to be novel, on the basis of comparisons with the dbSNP database of the National Center for Biotechnology Information (US) or with previous publications. The maps constructed in this study will serve as an additional resource for studies of complex genetic diseases and drug-response phenotypes to be mapped by linkage disequilibrium analyses. PMID- 12560876 TI - A double mutation (G11778A and G12192A) in mitochondrial DNA associated with Leber's hereditary optic neuropathy and cardiomyopathy. AB - We report a male patient with Leber's hereditary optic neuropathy (LHON) and hypertrophic cardiomyopathy. Besides a G11778A mutation in the ND4 gene of the mitochondrial DNA (mtDNA), one of the most common mutations in LHON patients, sequencing of total mtDNA revealed a G12192A mutation in the tRNA (His) gene that was recently noted to be a risk factor for cardiomyopathy. Because no case of LHON presenting with cardiomyopathy has been reported, the present finding suggests that the G12192A mutation caused cardiomyopathy as an additional symptom. In the present case, the double pathogenic mtDNA mutations may be associated either synergistically or concomitantly with two different clinical manifestations. PMID- 12560877 TI - Two novel mutations in the EPM2A gene in a Korean patient with Lafora's progressive myoclonus epilepsy. AB - The progressive myoclonus epilepsy of the Lafora type (LD; MIM 254780) is a rare autosomal recessive disorder characterized by epilepsy, myoclonus, progressive neurological deterioration, and the presence of periodic acid-Schiff-positive polyglucosan inclusions (Lafora bodies). Mutations in the EPM2A gene have recently been found to cause LD and about 30 or more mutations have been reported thus far. LD is relatively common in countries of the Mediterranean Basin, the Middle East, India, and Pakistan. Although a few sporadic cases with the typical LD phenotype have also been reported in the Far East including Korea and Japan, a recent effort to find mutations in Japanese LD families was not successful. In the present study, we report two novel mutations in a Korean girl with LD; a 1-bp insertion mutation (c.223insC; G75fsX107) in exon 1 and a missense mutation (c.559A>G; T187A) in exon 3 of the EPM2A gene. To our knowledge, this is the first report of a genetically confirmed case of LD in Koreans and also in the Far East. PMID- 12560878 TI - Assessment of lumbar spinal canal stenosis by magnetic resonance phlebography. AB - There is evidence to suggest that cauda equina intermittent claudication is caused by local circulatory disturbances in the cauda equina as well as compression of the cauda equina. We evaluated the role of magnetic resonance phlebography (MRP) in identifying circulatory disturbances of the vertebral venous system in patients with lumbar spinal canal stenosis. Extensive filling defects of the anterior internal vertebral venous plexus were evident in patients with lumbar spinal canal stenosis ( n = 53), whereas only milder abnormalities were noted in patients with other lumbar diseases ( n = 16) and none in normal subjects ( n = 13). The extent of the defect on MRP correlated with the time at which intermittent claudication appeared. In patients with lumbar spinal canal stenosis, extensive defects of the internal vertebral venous plexus on MRP were noted in the neutral spine position, but the defect diminished with anterior flexion of the spine. This phenomenon correlated closely with the time at which intermittent claudication appeared. Our results highlight the importance of MRP for assessing the underlying mechanism of cauda equina intermittent claudication in patients with lumbar spinal canal stenosis and suggest that congestive venous ischemia is involved in the development of intermittent claudication in these patients. PMID- 12560879 TI - Electrophysiological assessment of the carpal tunnel syndrome in hemodialysis patients: formula for predicting surgical results. AB - Nerve conduction studies performed between 1994 and 1996 on the median nerves of 204 hands of 113 patients [20 patients in hemodialysis (HD) with diabetic nephropathy, 27 without diabetes mellitus in HD for 10 years or more, 18 without diabetes mellitus in HD for less than 10 years, and 48 with diabetes mellitus not undergoing HD] were reviewed retrospectively to develop a discriminating tool that would better inform treatment selection for patients with carpal tunnel syndrome (CTS) in HD. Scatter diagrams were drawn from the results of this study, and a regression formula was obtained. Among the long-term HD patients without diabetic nephropathy, distal motor latency (DML) tended to increase without a significant concomitant reduction in motor nerve conduction velocity (MCV) in the forearm. In contrast, in HD patients with diabetic nephropathy, DML tended to increase and MCV was significantly lower despite shorter HD periods. The study showed that in HD patients, in whom the MCV(m/s) > 11[9 - DML(ms)], the surgical results might be excellent or good. The formula was tested against the results of the 30 surgical cases. The sensitivity of this formula was 0.84, and the specificity was 1.00. The formula produced a predictor of surgical results when used in conjunction with other criteria for CTS in HD patients, especially those with diabetic nephropathy. PMID- 12560880 TI - Histological and radiological analysis of autoclaved bone 2 years after extirpation. AB - We examined grafted distal femoral autoclaved bone radiologically and histologically 24 months after surgery. The patient was a 16-year-old boy with osteoblastic-type osteosarcoma in the distal part of the left femur. The patient received pre- and postoperative chemotherapy and underwent limb reconstruction surgery using an autoclaved autograft. He was forced to undergo hip disarticulation because of local recurrence in the soft tissue. Radiologically and histologically, we were able to detect newly formed bone at the site of the distal junction and surrounding the autoclaved autograft, although most of the autoclaved bone remained without substitution even 24 months after implantation. The layer of newly formed bone surrounding the autoclaved autograft was so thin that it seemed to be ineffective for weight-bearing. Drilling into the autoclaved autograft appeared to promote little bone regeneration inside the autoclaved autograft. A bone scintigram showed newly formed bone around the autoclaved autograft, but the scan tended to exaggerate such bone formation beyond that actually confirmed by histological examination. We should be careful when applying autoclaved bone for weight-bearing parts. PMID- 12560881 TI - Ankle arthrodesis combined with tibial lengthening using the Ilizarov apparatus. AB - We report our experience using the Ilizarov method to perform combined ankle arthrodesis and tibial lengthening in six patients (mean age 47 years; range 25 66 years). The average distraction length was 4.1 cm (range 1.1-6.8 cm), and the mean period of follow-up was 36 months (range 26-44 months). Three patients had active infection of the ankle. Four patients had undergone previous surgery, two of which were primary ankle arthrodeses. We performed compression-distraction in three patients and bone transport in three. In the compression-distraction group, the mean length gained was 1.9 cm, the mean external fixation index (EFI) was 144 days/cm, and the mean external fixation time was 246 days. In the bone transport group, the mean length gained was 6.2 cm, the mean EFI was 35.4 days/cm, and the mean external fixation time was 233 days. All cases achieved a good clinical result with a solid ankle arthrodesis and no infection, deformity, or need for additional support. The Ilizarov method may be practically applied for ankle arthrodesis, especially in complicated cases. The EFI and external fixation time tended to increase for patients with a length gain of 3 cm or less. PMID- 12560882 TI - Treatment outcome of two-stage revision total hip arthroplasty for infected hip arthroplasty using antibiotic-impregnated cement spacer. AB - Infected hip prosthesis, a serious complication of primary total hip arthroplasty (THA), can have severe consequences. We report the treatment outcome of two-stage revision THA for infected hip arthroplasty, including hemiarthroplasty, using an antibiotic-impregnated cement spacer for the interval between the first and second stages. Between 1996 and 2000 we performed this procedure on nine hips in eight patients. Cementless revision THA was performed as the second-stage procedure. Bone defects were restored with frozen allografts. The outcome was evaluated using the hip score of the Japanese Orthopaedic Association (JOA hip score). The mean duration of follow-up was 35.7 months (range 10-55 months). The mean JOA hip score at follow-up improved from 30.1 (range 10-74) to 73.2 (24-96). The mean interval between the first and second stages was 10.1 weeks (range 6-19 weeks). Eight of the nine hips achieved a successful outcome. One hip, with methicillin-resistant Staphylococcus aureus infection, experienced recurrence 4 months after revision THA. This patient was successfully treated 14 months after the first revision THA with a second two-stage procedure using a vancomycin- and arbekacin-impregnated cement spacer and beads. These results suggest that two stage revision THA using an antibiotic-impregnated cement spacer is a useful technique for treating infected hip arthroplasty. PMID- 12560883 TI - Quality of life after several treatments for osteoarthritis of the hip. AB - The purpose of this study was to examine quality of life (QOL) after a rotational acetabular osteotomy (RAO), primary total hip arthroplasty (THA), or conservative treatment (Conservative). A series of 147 patients with osteoarthritis of the hip were grouped according to treatment into the RAO group ( n = 62), THA group ( n = 40), and Conservative group ( n = 45) (treated with medication and rehabilitation alone). The mean age of the RAO group was 40 years, the THA group 54 years, and the Conservative group 50 years. The mean follow-up periods were 3.6, 3.9, and 3.3 years, respectively. All patients were female. As a control group 609 volunteers participating in a town mass screening program were recruited. QOL was determined using the Nottingham Health Profile (NHP). Each area of QOL was evaluated separately in all of the treatment groups at less than 1 year after therapy, at more than 1 year but less than 5 years, and at more than 5 years. With regard to QOL at less than 1 year after therapy, the Conservative group had significantly higher scores for energy, pain, and emotional reaction than the Control group. With regard to physical mobility, all of the treatment groups showed significantly higher scores than the Control group. With regard to QOL at more than 1 year but less than 5 years, the RAO and THA groups showed significantly higher scores than the Control group regarding physical mobility. Regarding pain, the RAO and Conservative groups showed significantly higher scores than the Control group. With regard to QOL at more than 5 years, for physical mobility all of the treatment groups showed significantly higher scores than the Control group. For pain the RAO group showed significantly higher scores than the Control group. NHP is a useful means for evaluating QOL in patients treated for osteoarthritis of the hip. PMID- 12560884 TI - Cervical muscle strength after laminoplasty. AB - To determine changes in cervical muscle strength after laminoplasty and to evaluate the relation between muscle strength and neck pain, we measured maximum isometric muscle strength using a handheld dynamometer. We also investigated neck pain before surgery and every month after surgery in 21 subjects who had undergone French-door laminoplasty. Muscle strength decreased particularly 1 month after surgery, the extensor muscles being affected more than the flexors. The strength of both muscle groups increased gradually; and at 1 year after surgery they had regained their presurgical status. All of the subjects complained of severe neck pain after 1 month. Their complaints began to decrease a few months after surgery, although they were still present in nine patients at 1 year after surgery. The correlation between muscle strength and neck pain was strongly negative for extension and flexion in men and for extension in women. No correlation was found between flexor muscle strength and neck pain in women. The extension/flexion ratio was significantly high in the neck pain group 1 year after surgery. Our results suggest that symptoms within a few months after surgery are due to surgical trauma to the soft tissues but that chronic neck pain derives from an imbalance of the two muscle strengths. PMID- 12560885 TI - Spinal penetration index: new three-dimensional quantified reference for lordoscoliosis and other spinal deformities. AB - We studied and conceptually analyzed a retrospective case series of patients with airway compression due to an anterior vertebral body protrusion. The goal was to describe the pathology, methods of management, and a new concept for quantifying deformity. Case reports have been published on this pathology, but there has been no case series to date. In this study 18 patients with ages ranging from 7.3 to 18.0 years had thoracic lordoscoliosis due to a variety of etiologies; most ( n = 10) had a neuromuscular disorder. Following treatment, which most commonly was anterior subtotal subperiosteal vertebral body resection followed by posterior instrumentation and arthrodesis, atelectasia disappeared and any abnormal blood gases normalized; however, the effect on vital capacity was variable. Based on computed tomographic studies, the concept of the deformity as an endothoracic vertebral hump was developed and quantified. Study of this series of patients with compression of the airway due to vertebral body protrusion into the thorax provided the opportunity to describe treatment, define a new concept (the spinal penetration index), and make general recommendations about the management of both the endothoracic hump and the exothoracic rib hump. PMID- 12560886 TI - Biomechanical study of human cadaveric lumbar spine reinforced by newly developed hydroxyapatite bone cement. AB - The compression strength of the lumbar spine reinforced by newly developed hydroxyapatite (HA) bone cement was evaluated using a mechanical testing machine. Sixteen cadaveric lumbar vertebrae obtained from nine subjects (five men, four women) were used. The specimens were randomly divided into two groups. In group A ( n = 8), HA bone cement was injected into the vertebral body through curetted pedicles using specially designed needles and then pushed into the vertebral body by the surgeon's finger, simulating open surgery. In group B ( n = 8) the cement was injected using 16-gauge Ostycut biopsy needles via the pedicles through both sides, simulating percutaneous injection. The initial ultimate compression strength of the specimens was 28.6 +/- 13.4 MPa in group A and 25.2 +/- 12.6 MPa in group B. The value after reinforcement was 35.6 +/- 12.9 MPa in group A and 30.4 +/- 14.8 MPa in group B. There was no significant difference between the ultimate strength of the intact specimen and that after reinforcement. The present study demonstrated biomechanical characteristics of vertebral body fractures reinforced with newly developed HA bone cement. PMID- 12560887 TI - FDG-PET imaging of lower extremity muscular activity during level walking. AB - We analyzed muscular activity of the lower extremities during level walking using positron emission tomography (PET) with (18)F-fluorodeoxyglucose ((18)F-FDG). We examined 17 healthy male subjects; 11 were assigned to a walking group and 6 to a resting group. After (18)F-FDG injection, the walking group subjects walked at a free speed for 15 min. A whole-body image was then obtained by a PET camera, and the standardized uptake ratio (SUR) was computed for each muscle. The SUR for each muscle of the walking group was compared with that for the corresponding muscles in the resting group. The level of muscular activity of all the muscles we examined were higher during level walking than when resting. The activity of the lower leg muscles was higher than that of the thigh muscles during level walking. The muscular activity of the soleus was highest among all the muscles examined. Among the gluteal muscles, the muscular activity of the gluteus minimus was higher than that of the gluteus maximus and gluteus medius. The concurrent validity of measuring muscular activity of the lower extremity during level walking by the PET method using (18)F-FDG was demonstrated. PMID- 12560888 TI - Vascularized synovial flap promoting regeneration of the cryopreserved meniscal allograft: experimental study in rabbits. AB - The purpose of this investigation was to evaluate whether a vascularized or free synovial flap or a fibrin clot can promote regeneration of meniscal allograft in the rabbit. Seventy-eight mature New Zealand white rabbits were used. The harvested medial meniscus for the allotransplantation was frozen and stored at 80 degrees C for 2 weeks. After resecting the medial meniscus, an allogenic meniscus was transplanted in the anatomical position (group A). The surface of the graft was covered by a vascularized synovial flap (group B), a free synovial flap (group C), or a fibrin clot (group D). The animals were killed 4, 6, 8, 12, and 16 weeks after transplantation, and the transplants were examined by gross inspection, histology, and microangiography. Connective tissue infiltration into the matrix of the graft was found to begin at 6 weeks (2/5 menisci) and to be complete at 8 weeks (5/5 menisci) in group B, whereas it began at 8 weeks (1/5) in group A. The newly formed repair tissue developed from the synovial tissue and consisted of connective tissue at the beginning and fibrocartilage later. The fibrocartilage had appeared at 8 weeks (3/5) in group B but not yet in group A (0/5). A free flap or fibrin clot coverage resulted in delayed revascularization compared to a vascularized synovial flap, but both tended to achieve faster revascularization than the controls. We concluded that regeneration of allografted meniscus with a vascularized synovial flap occurs earlier than under other conditions. Thus, allografts with synovial implantation may be considered for management of the meniscectomized knee. PMID- 12560889 TI - Motor nerve conduction analysis of double crush syndrome in a rabbit model. AB - The double crush syndrome was proposed by Upton and McComas in 1973, but there are still many unclear points regarding its mechanism. We propose a model that enables electrophysiological study of the same nerve from the same individual over time. We employed 29 rabbits and used their sciatic nerves. (Four rabbits died from infection, and in nine rabbits we could not record the M-wave because the electrodes were off during the course of the study.) Sixteen rabbits were grouped into six experimental groups and one control group. Bipolar stimulus electrodes were buried at three locations in their sciatic nerves. The two areas between the proximal and intermediate electrodes and between the intermediate and distal electrodes were defined as the "nerve proximal part" and "nerve distal part," respectively. An electrical stimulus was applied via each electrode, and the muscle action potential (M-wave) was recorded from the gastrocnemius. A narrow tube was inserted in the nerve proximal part and a wide tube in the nerve distal part. A decrease in the motor nerve conduction velocity (MCV) was observed in the nerve proximal part (narrow tube) followed by a decrease in MCV in the nerve distal part (wide tube). A wide tube was then inserted in the nerve proximal part and a narrow tube in the nerve distal part. A decrease in MCV was observed in the nerve distal part (narrow tube) followed by a decrease in MCV in the nerve proximal part (wide tube). We believe that these results are important findings in regard to the double crush syndrome (DCS) hypothesis and the reversed DCS hypothesis. PMID- 12560890 TI - Changes of fiber type ratio and diameter in rabbit skeletal muscle during limb lengthening. AB - Changes in the fiber-type ratio and diameter during limb lengthening in 10 adult rabbits were studied using histochemical techniques. Changes in the ratio and diameter of muscle fibers (classified as type 1, 2A, or 2B fibers) in tibialis anterior muscles were examined after 20% gradual distraction of the tibia. There was an increase in the number of type 1 fibers and a decrease in type 2B fibers after tibial lengthening. Moreover, the average diameter of the type 1 fibers increased, whereas that of the type 2B fibers decreased. The diameters of muscle fibers measured immediately after completion of the lengthening showed a tendency to recover to normal levels within 1 month; however, the ratio profile of the muscle fibers changed both immediately and 1 month after lengthening. The above results demonstrate that stretched skeletal muscle adapts differently to bone lengthening according to the type of muscle fibers present, resulting in qualitative changes in the fiber-type profile. PMID- 12560891 TI - Thalidomide blocking of particle-induced TNFalpha release in vitro. AB - Tumor necrosis factor-alpha (TNFalpha) and interleukin-6 (IL-6), pleiotropic cytokines with osteotropic activities, are produced by multiple cells in the skeletal tissue, including macrophages and osteoblasts. They are thought to be pivotally involved in pathological bone resorption, such as that seen with aseptic loosening. Thalidomide is reported to have antiinflammatory, immunomodulatory effects in a number of inflammatory diseases. We investigated the effect of thalidomide on titanium (Ti) particle-induced TNFalpha and IL-6 production by both human macrophage U937 and osteoblast MG-63 cell lines. They were stimulated with 1 x 10(7) Ti particles/ml and treated simultaneously with or without various concentrations of thalidomide (from 2.5 ng/ml to 25 microg/ml) for 24, 48, or 72 h. Cell viability and proliferation were measured. TNFalpha and IL-6 in the supernatant of the culture media were also analyzed with an enzyme linked immunosorbent assay. We found that with a concentration of thalidomide of less than 2.5 microg/ml the viability of the two cell lines did not differ significantly from that of controls treated simultaneously with 1 x 10(7) Ti particles/ml. Cell proliferation was inhibited to some extent when they were treated with thalidomide 2.5 microg/ml co-cultured with 1 x 10(7) Ti particles/ml. Thalidomide treatment was found to inhibit TNFalpha production in a dose-dependent manner in human macrophages exposed to Ti particles. At the clinically achievable drug dose of 2.5 microg/ml, 34.4% TNFalpha inhibition occurs. Thalidomide had no effect on IL-6 secretion in these cultures. These data support the idea that thalidomide may have potential for treating prosthetic loosening in humans. PMID- 12560892 TI - Existence of brain-derived neurotrophic factor and vanilloid receptor subtype 1 immunoreactive sensory DRG neurons innervating L5/6 intervertebral discs in rats. AB - The rat L5/6 intervertebral disc is innervated by L1 to L6 dorsal root ganglia (DRGs). T13 to L2 DRGs innervate the L5/6 intervertebral disc through paravertebral sympathetic trunks, whereas L3 to L6 DRGs directly innervate through sinuvertebral nerves on the posterior longitudinal ligament. The presence of substance P (SP)-immunoreactive (ir) and calcitonin gene-related peptide (CGRP ir) sensory nerve fibers on the lumbar intervertebral disc has been established. SP and CGRP are markers of sensory neurons mainly involved with pain perception. The existence of SP-ir and CGRP-ir DRG neurons innervating the L5/6 intervertebral disc has been also demonstrated. Brain-derived neurotrophic factor (BDNF), which exists mainly in the small DRG neurons, plays an important neuromodulatory role in inflammatory conditions. Vanilloid receptor subtype 1 (VR1) in the DRG neurons and spinal dorsal horn is a channel that appears to confer responsiveness to heat and chemical stimuli. The presence of BDNF-ir and the VR1-ir DRG neurons innervating the L5/6 intervertebral disc has not. In this study of DRG neurons innervating the L5/6 intervertebral disc, the proportions of BDNF-ir in L1, L2, L3, L4, and L5 DRG neurons were 14%, 12%, 12%, 12%, and 13% and the proportions of VR1-ir L1, L2, L3, L4, and L5 DRG neurons were 10%, 8%, 24%, 19%, and 23%, respectively. Under physiological conditions in rats these neurons may transmit inflammatory and burning pain of the L5/6 intervertebral disc. PMID- 12560893 TI - Histological and electrophysiological assessment of the results of primary and secondary neurorrhaphy in a rabbit model. AB - The effect of the timing of neurorrhaphy on neuromuscular function was studied. The gastrocnemius neuromuscular units of 22 rabbits were used, with repairs performed immediately and 3 weeks after nerve transection. At the end of 3 months, electrophysiological and histological results were compared. The study found that there was a higher rate of neuromuscular function return in primarily sutured nerves (88.0%) than in nerves repaired by secondary suture (63.0%). The histological examination also confirmed a better preserved microscopic structure of the primary repair group. PMID- 12560894 TI - Repair of full-thickness cartilage defects using liposomal transforming growth factor-beta1. AB - Transforming growth factor-beta1 (TGFbeta1) is a well- known, potent growth factor implicated in both in vitro and in vivo chondrogenesis. Liposomes have been employed as a drug delivery system to promote the efficient use of drugs. The objective of this study was to demonstrate that a single injection of liposomal TGFbeta1 has an accelerating effect on the repair of an articular cartilage defect. Full-thickness articular cartilage defects were prepared on the patellar grooves of the femurs in knee joints of Japanese white rabbits. One week after surgery, various reagents including liposomal TGFbeta1, free TGFbeta1, and phosphate-buffered saline were injected into the operated knee joints. At 3 weeks after surgery the specimens obtained from the lesions were evaluated histologically, and the glycosaminoglycan content was quantified. Histological examination revealed that the defects were filled with thicker fibrous cartilage and showed more intense metachromatic staining in the liposomal TGFbeta1 group than in the other groups. The glycosaminoglycan content of the repair tissue was also significantly higher in the liposomal TGFbeta1 group than in the other groups. This study indicated that the intraarticular injection of liposomal TGFbeta1 could accelerate the early-stage repair of full-thickness articular cartilage defects. PMID- 12560895 TI - Enhanced expression of Runx2/PEBP2alphaA/CBFA1/AML3 during fracture healing. AB - The cause of the dramatic increase in expression of the osteopontin gene during fracture healing was studied in a mouse experimental model. Semiquantitative reverse transcription-polymerase chain reaction, Northern blotting, and in situ hybridization analysis showed that the enhanced expression took place prior to callus formation. The change in the expression pattern of collagenous and noncollagenous bone matrix proteins in addition to Ets-1 and Runx2, major transcription factors of osteopontin, were examined and compared to that of osteopontin. Although Ets-1 expression showed no significant change during fracture healing, enhanced expression of Runx2 corresponding to that of osteopontin was observed. Furthermore, in situ hybridization demonstrated that osteopontin-expressing cells also express the Runx2 gene. The results indicated the possibility that Runx2 is a major regulator of osteopontin during fracture healing. PMID- 12560896 TI - Lumbar epidural abscess causing septic shock: case report. AB - This case report describes a lumbar epidural abscess that caused septic shock. A 48-year-old woman who developed a high temperature was diagnosed as having a respiratory tract infection. She became unconscious the next day and was transferred to the intensive care unit for monitoring and treatment. She complained of increasing lower back pain as consciousness improved. Magnetic resonance images of the lumbar spine showed an abscess shadow between the vertebral body and dura mater at the level of L3-S1. Surgical drainage of the epidural abscess was performed as an emergency procedure, and the patient recovered immediately. The primary source of the epidural abscess was probably the respiratory tract infection, which spread to the epidural space through hematogenous dissemination. The initial treatment of the epidural abscess was rest and antibiotics, but surgical treatment was needed to save the patient because of septic shock. PMID- 12560897 TI - Osteochondroma in the lumbar spinal canal causing sciatic pain: report of two cases. AB - A search of the English-language medical literature found only two cases in which expansion of an osteochondroma into the lumbar spinal canal caused sciatica. We report another two cases of spinal nerve root compression by solitary lumbar spinal canal osteochondromas: in a 56-year-old man and a 55-year-old woman with no history of hereditary multiple exostoses. Osteochondromas compressing the spinal nerve root were seen at the inferior articular processes of the lumbar vertebrae by computed tomography (CT), three-dimensional reconstruction of CT scans, myelography, and magnetic resonance imaging. The symptoms disappeared after surgical removal of the lesions. Histopathologic examination confirmed the diagnosis of benign osteochondroma. PMID- 12560898 TI - Distraction osteogenesis for treatment of bone loss in the lower extremity. AB - Distraction osteogenesis has been widely utilized to treat leg length discrepancy, deformity, nonunion, osteomyelitis, and bone loss. It has been found that most difficult conditions can be resolved with this method. This paper introduces the usefulness of distraction osteogenesis for the treatment of nonunion and bone loss after tumor excision. The procedure was found to be extremely effective for the treatment of nonunion accompanied by bone loss and infected nonunion. In particular, the Ilizarov method offers a simultaneous solution for several problems related to nonunion, such as instability, infection, deformity, bone loss, joint contracture, limb length discrepancy, and skin defects. In addition, distraction osteogenesis is safe, useful, and efficient for the treatment of bone loss even after tumor excision. Joint preservation and reconstruction by means of distraction osteogenesis is the most conservative limb-saving surgery available at present. Furthermore, distraction osteogenesis can provide natural limb regeneration. PMID- 12560899 TI - Postoperative complications of pulmonary resection after platinum-based induction chemotherapy for primary lung cancer. AB - PURPOSE: We investigated the postoperative complications that developed in patients who underwent surgery after induction chemotherapy (IC) for primary lung cancer. METHODS: Twenty-seven patients underwent surgery after receiving IC; for advanced non-small cell lung cancer in 16, and for small cell lung cancer in 11. All patients were given the platinum-based chemotherapy regimen. RESULTS: Lobectomies were performed for 18 patients, bilobectomies for 4, pneumonectomies for 2, and partial resections or segmentectomies for 3. There were two postoperative deaths; one caused by adult respiratory distress syndrome (ARDS) and one caused by respiratory failure, resulting in a mortality rate of 7.4%. The postoperative complications included sputum retention in six patients, ARDS in two, anastomotic dehiscence after bronchoplasty in one, and pneumonia in one, resulting in 44.4% morbidity. The morbidity of patients who had received IC (IC group) was higher than that of a comparative group of 560 who underwent lung resection without IC during the same period (non-IC group), but the difference was not significant (44.4% vs 22.6%; P = 0.16). Both ARDS and bronchial insufficiency occurred more frequently in the IC group than in the non-IC group, but the differences were not significant ( P = 0.25). CONCLUSIONS: These findings indicate the feasibility of treating primary lung cancer with IC followed by surgery as long as a cautious operative procedure is used and careful postoperative management is given, paying particular attention to the risk of ARDS and bronchial complications. PMID- 12560900 TI - Difference in the impairment of vital capacity and 6-minute walking after a lobectomy performed by thoracoscopic surgery, an anterior limited thoracotomy, an anteroaxillary thoracotomy, and a posterolateral thoracotomy. AB - PURPOSE: Postoperative vital capacity (VC) and the 6-min walking (6MW) test were used to compare the differences in impairment of the pulmonary function and walking capacity in patients undergoing a lobectomy by video-assisted thoracoscopic surgery (VATS), an anterior limited thoracotomy (ALT), an anteroaxillary thoracotomy (AAT), or a posterolateral thoracotomy without muscle sparing (PLT). METHODS: The study was a retrospective analysis. Lung cancer patients who underwent a lobectomy by VATS, ALT, AAT, or PLT (28 in each group) were matched by sex and age (+/-5 years). VC was measured before surgery and at 1, 2, 4, 12, and 24 weeks after surgery. The distance covered during the 6MW test (6MWD) was measured before surgery and in a postoperative test 1 week after surgery. RESULTS: Compared with the VATS, ALT, and AAT groups, PLT patients showed a significant impairment of VC from 1 to 24 weeks after surgery ( P < 0.05 0.001) and also a significant impairment of 6MWD 1 week after surgery ( P < 0.01 0.001). The AAT group showed a significant impairment of 6MWD 1 week after surgery compared with the VATS and ALT groups ( P < 0.001 and P < 0.05, respectively). There was no significant difference in the impairment of either VC or 6MWD between VATS and ALT. CONCLUSIONS: The PLT without a muscle sparing procedure therefore cannot be recommended for general lung cancer surgery because of the impairment of both walking capacity and pulmonary function which continues long after surgery. VATS and ALT are better procedures than AAT regarding the recovery of walking capacity early after surgery. VATS and ALT are similar to each other regarding the impairment of pulmonary function and walking capacity after surgery. PMID- 12560901 TI - Factors affecting the early mortality of patients with nontraumatic colorectal perforation. AB - PURPOSE: We attempted to identify the factors associated with the early mortality of patients with nontraumatic colorectal perforation. METHODS: Eighty patients who underwent surgery for nontraumatic colorectal perforation between May 1986 and December 1999 were retrospectively reviewed. Age, sex, cause of perforation, duration of symptoms, associated preoperative septic shock, concomitant disorders (including cardiac disease, chronic obstructive pulmonary disease, hemodialysis, and steroid treatment), operative findings (such as the site of perforation and the degree of peritonitis), and results of preoperative laboratory blood tests (such as the white blood cell count and platelet count) were analyzed for their association with early outcome using univariate and multivariate analyses. RESULTS: Fourteen of the 80 patients died during hospitalization. According to the univariate analysis, advanced age, preoperative septic shock, concomitant disabling cardiac disease, hemodialysis, diffuse peritonitis, and a low preoperative platelet count were more frequent in the patients who died during hospitalization. According to the logistic regression analysis, preoperative septic shock (odds ratio 8.443, 95% confidence interval (CI) 1.625-43.873), concomitant end-stage renal failure (odds ratio 13.641, 95% CI 1.643-113.244), and diffuse peritonitis (odds ratio 13.212, 95% CI 1.441-121.102) were the most significant factors related to in-hospital mortality. CONCLUSION: Early diagnosis before the patient's general condition deteriorates is a key to improving the early mortality associated with nontraumatic colorectal perforation, especially in patients with concomitant end-stage renal failure. PMID- 12560902 TI - Efficacy of absorbable clips compared with metal clips for cystic duct ligation in laparoscopic cholecystectomy. AB - PURPOSE: This study was conducted to examine the usefulness and safety of absorbable clips in laparoscopic cholecystectomy (LC). METHODS: We retrospectively compared the clinical data of 328 patients who underwent LC using absorbable clips for cystic duct ligation and 444 patients who underwent LC using metal clips for cystic duct ligation. RESULTS: The mean operative time in the absorbable clip group was significantly shorter than that in the metal clip group. The mean intraoperative blood loss and hospital stay were not significantly different between the two groups. The rate of conversion to open laparotomy in the absorbable clip group was significantly higher than that in the metal clip group, at 6.7% vs 2.3%, respectively. The most common reason for conversion was difficult adhesions and inflammation around the gallbladder. The morbidity of the metal clip group was significantly higher than that of the absorbable clip group, at 8.4% vs 4.0%, respectively. The incidence of major postoperative complications requiring laparotomy did not differ between the two groups. There were no deaths in the absorbable clip group, but 1 of the 444 patients (0.23%) in the metal clip group died from disseminated intravascular coagulation on postoperative day 3. CONCLUSION: The results of this study suggest that absorbable clips are as safe and effective as standard metal clips for vessel and duct ligation in LC. PMID- 12560903 TI - Thyroid hormone supplementation in sepsis: an experimental study. AB - PURPOSE: The levels of thyroid hormones decrease in septic, critically ill, and major trauma patients who show a bad prognosis. The aim of this study was to investigate the effects of thyroid hormone supplementation on the sepsis criteria and mortality in an experimental sepsis model. METHODS: Forty-eight Wistar-Albino rats were divided into four experimental groups. A cecum ligation and puncture was used as a sepsis model. Group I: sham; group II: a cecum ligation and puncture (control); group III: a cecum ligation and puncture +T3 (0.4 microg/100 g body weight/day) intraperitoneally; group IV: a cecum ligation and puncture +T4 (1 500 ng/100 g body weight/day) intramuscularly. The PCO(2), leukocyte count, peritoneal bacterial content, neutropenia, and mortality rates were assessed in all groups. RESULTS: The PCO(2) levels decreased only in the control group ( P < 0.05). The leukocyte counts were lower in the therapy groups ( P < 0.05). The rate of neutropenia was seen much more in the control group ( P < 0.05). The peritoneal bacteria content decreased in the therapy groups ( P < 0.05). The mortality rate significantly decreased in the thyroid hormone-treated animals ( P < 0.05). CONCLUSIONS: Thyroid hormone supplementation in septic rats had a beneficial effect on the sepsis criteria and also resulted in a lower rate of mortality. PMID- 12560904 TI - Inhibition of heme oxygenase ameliorates sepsis-induced liver dysfunction in rats. AB - PURPOSE: The disintegration of heme produces carbon monoxide (CO), a known vasodilator, which is catalyzed by heme oxygenase (HO). This study aimed to clarify the effect of HO inhibition on septic rat livers using two types of HO inhibitors; Sn-protoporphyrin (Sn-PP) and Zn-protoporphyrin (Zn-PP). METHODS: Sepsis was induced in male Sprague-Dawley rats by cecal ligation and puncture (CLP). Either NaOH or HO inhibitors were injected intraperitoneally; first 18 h prior to CLP, then immediately after CLP. The animals were killed 12 and 24 h after CLP and the liver tissue and plasma were harvested. RESULTS: Using Northern blotting, we found that mRNA of the stress-inducible isozyme, HO-1, was dramatically induced 12 h after CLP. Administering the HO inhibitors, Sn-PP and Zn-PP (5 micromol/kg), induced a significant inhibition of the elevation of aspartate aminotransferase plasma levels, the elevation of cyclic guanosine monophosphete (cGMP) in the liver tissue, and the increase in the sinusoidal space ratio, 24 h after CLP. Both Sn-PP and Zn-PP decreased the mortality rate 24 h after CLP compared with normal saline. CONCLUSIONS: CO produced by excessively induced HO-1 after CLP promotes an immoderate dilation of the sinusoidal space through the up-regulation of cGMP, resulting in liver dysfunction. Therefore, administering HO inhibitors at appropriate doses could be beneficial for the amelioration of sepsis-induced liver dysfunction. PMID- 12560905 TI - Possible involvement of antitumor immunity in the eradication of colon 26 induced by low-voltage electrochemotherapy with bleomycin. AB - PURPOSE: The antitumor efficiency of electrochemotherapy using chemotherapeutic agents and high-voltage electric pulse has been reported. This study was done to define the precise nature of the involvement of antitumor immunity in the regression of tumor nodules in electrochemotherapy, and to evaluate the effectiveness of using low-voltage electroporation. METHODS: Balb/c mice and Balb/c nu/nu nude mice were inoculated subcutaneously with Colon 26 cells or Meth A cells. Electrochemotherapy using bleomycin and low-voltage electroporation (CUY21) was performed as a treatment against tumor nodules. RESULTS: Colon 26 tumors were eradicated in the mice given an intratumor (i.t.) injection of 500 microg bleomycin followed by treatment with electric fields ranging from 50 to 150 V/cm, with complete response rates ranging from 80% to 100%. The mice rejected inoculations of rechallenged Colon 26 cells, but not Meth A cells. In the Balb/c nu/nu nude mice, complete regression of the tumor was not seen after electrochemotherapy under the same therapeutic conditions that resulted in almost complete cure in the Balb/c mice. CONCLUSION: Our results suggest that the generation of T-cell-dependent, tumor-specific protective immunity might be involved in the process of tumor nodule regression in low-voltage electrochemotherapy. PMID- 12560906 TI - Salvage surgery for primary non-Hodgkin's lymphoma of the thyroid gland with histopathological complete response to radio-chemotherapy: report of a case. AB - A 74-year-old woman was admitted to a local hospital for investigation of a rapidly growing mass in her neck, and exertional dyspnea. An open biopsy confirmed a diagnosis of non-Hodgkin's lymphoma of the thyroid (NHLT), of a diffuse large cell type. The patient was referred to our department for radio chemotherapy for stage I E NHLT. She was given radiotherapy in the form of 40 Gy radiation directed at her neck and superior mediastinum, with one course of chemotherapy using cyclophosphamide, adriamycin, vincristine, and prednisolone (CHOP). After radio-chemotherapy, the tumor was obviously smaller. Because the patient refused further chemotherapy, she underwent salvage surgery, after being sufficiently advised, and with her informed consent. Histological examination of the removed thyroid tissue showed that the radio-chemotherapy had produced a complete response. Thus, we believe that an open biopsy should be performed early to confirm the diagnosis of lymphoma histologically and to determine the degree of malignancy. We also stress the fact that NHLT is presently most effectively treated by radiotherapy combined with several courses of CHOP chemotherapy. The role of surgery in the treatment of NHLT is diminishing. PMID- 12560907 TI - Multiple thymoma with myasthenia gravis: report of a case. AB - A resected case of multiple thymoma is reported. The patient was a 47-year-old man with myasthenia gravis of Ossermann IIB type. Multiple thymomas were detected on a chest computed tomography scan and an extended thymothymectomy was performed. There were two separate thymomas in the anterior mediastinum: one measuring 60 x 60 mm and another measuring 25 x 25 mm in diameter, and both tumors pathologically consisted of predominantly lymphocytic forms. PMID- 12560908 TI - Techniques for treating aberrant arteries during resection of pulmonary sequestration by video-assisted thoracic surgery: report of two cases. AB - We describe the techniques we used for treating aberrant arteries during resection of pulmonary sequestration by video-assisted thoracic surgery (VATS) in two patients. In patient 1, the aberrant artery was transected after securing six rows of staples with a knifeless vascular endostapler. In patient 2, the aberrant artery was cut after ligation with special forceps that designed by one of us (S.K.) to push a knot, tied outside the body, into the thoracic cavity, then ligate the suture. Left lower lobectomy and right basal segmentectomy were both successfully performed by these methods. These two cases are reported to show that VATS lobectomy is a feasible and minimally invasive technique of treating pulmonary sequestration and other diseases of the lung. PMID- 12560909 TI - Protruding thrombus in the left atrium found 7 years after percutaneous transvenous mitral commissurotomy: report of a case. AB - A 50-year-old man was transferred to our hospital for investigation of cerebellar infarction, thought to have been caused by cardiac thromboembolism. We assumed that the cardiac thromboembolism had occurred as a late complication of a percutaneous transvenous mitral commissurotomy (PTMC) performed 7 years earlier. An echocardiogram and thoracic computed tomography revealed a protruding thrombus in the left atrium and an emergency operation was performed. The protruding thrombus was found to originate from the scar that penetrated into the intra atrial muscular septum caused by the PTMC. After removing the thrombus, the scar was covered with normal endothelium and the mitral valve was replaced with a 27 mm St. Jude Medical prosthetic valve. We think that the thromboembolism was caused by mitral valve restenosis, atrial fibrillation, and endothelial injury in the interatrial septum during PTMC. Therefore, long-term follow-up and appropriate medication is recommended after PTMC, since restenosis and thrombosis are likely to occur. PMID- 12560910 TI - Surgical treatment of chronic intestinal pseudo-obstruction: report of three cases. AB - We report the cases of three patients with chronic intestinal pseudo-obstruction (CIP) whose quality of life (QOL) was improved by palliative surgery in combination with home parenteral nutrition (HPN). Loop enterostomy and shortening of the gastrointestinal tract was performed with good results in all three patients to relieve abdominal distension that was progressive despite treatment with prokinetic drugs. Oral intake was also improved after surgical treatment in one patient who underwent massive resection of the jejunoileum. HPN maintained a satisfactory nutritional state in all patients. These three case reports serve to demonstrate that surgical intervention to improve QOL should be considered for patients with CIP if medication is not effective and symptoms are progressive. PMID- 12560911 TI - A small, incidentally detected pancreatic somatostatinoma: report of a case. AB - We report an asymptomatic 72-year-old woman with a small, incidentally detected, pancreatic somatostatinoma. The tumor, measuring 1 cm in diameter, showed a hypervascular pattern of contrast enhancement on computed tomography, and was found angiographically to receive a blood supply from the posterior superior pancreaticoduodenal artery. The results of preoperative hormonal assays all were normal. No assay for somatostatin was performed. No abnormality in either the pituitary or parathyroid was found. We thus considered the tumor to be a sporadic, nonfunctioning endocrine cell tumor, and enucleation was carried out. As some tumor cells in the resected specimen showed immunoreactivity for somatostatin, a diagnosis of somatostatinoma was made. Therefore, the possibility of somatostatinoma should be kept in mind when making a differential diagnosis of pancreatic endocrine tumors in cases where even a small hypervascular tumor is detected on enhanced computed tomography. PMID- 12560912 TI - Bouveret's syndrome as a rare complication of cholecystolithiasis: report of a case. AB - Bouveret's syndrome, which is gastric outlet obstruction caused by a gallstone in the duodenum or pylorus, is a very rare complication of gallstone disease. It occurs most commonly in women (65%), with a median age of 68.6 years. This disorder is usually treated by surgery, but it has also been successfully treated by endoscopy, with or without extracorporeal shock wave lithotripsy. The mortality rate has improved to 12% in recent years. Herein we report the case of a 76-year-old woman with Bouveret's syndrome, and review the literature on this unusual entity. PMID- 12560913 TI - Hydatid cystic disease of the soft tissues with pulmonary and hepatic involvement: report of a case. AB - Hydatid disease, which is caused by the Echinococcus granulosis parasite, is endemic in certain parts of the world where close contact between humans and sheep exists. The disease most commonly involves the liver and other solid organs. This report describes the case of a teenage girl with hydatid disease involving the right axilla and various other sites concomitantly. The patient was admitted to hospital with a mass in the right axillary region, and further examinations revealed that the liver and right lung were also involved. We report this case to emphasize that hydatid disease should be considered in the differential diagnosis of patients from endemic areas who present with soft tissue masses. PMID- 12560914 TI - A successful right axillary artery graft to repair a ruptured axillary artery due to the involvement of lymphoma: report of a case. AB - Diffuse large B-cell lymphoma is a very common, highly invasive lymphoma, which typically presents as a rapidly enlarging symptomatic mass with local compression of vessels or airways, and often is involved with the peripheral nerves and the destruction of bone. Vascular invasion is extremely rare. We herein describe the case of a successfully treated 42-year-old man who presented with massive bleeding due to a rupture of the axillary artery and vein involvement due to lymphoma. PMID- 12560915 TI - Acute peritonitis caused by intraperitoneal rupture of an infected urachal cyst: report of a case. AB - Embryologically, the urachus is the tubular structure that connects the dome of the bladder to the umbilicus. Incomplete obliteration of the urachal lumen results in several anomalies. The most common urachal abnormality is the urachal cyst and, while intraperitoneal rupture of an infected urachal cyst is very rare, acute peritonitis resulting from intraperitoneal rupture is the most dangerous of all complications associated with urachal anomalies. We report the case of an 80 year-old woman who underwent an emergency laparotomy for lower abdominal pain and signs of acute peritonitis, which revealed intraperitoneal rupture of an infected urachal cyst. Infected urachal cysts with intraperitoneal rupture are often misdiagnosed as a common acute abdomen and result in emergency exploratory laparotomy. These patients should be managed by complete excision of the urachal remnant to prevent any malignant change occurring, as malignant changes have been reported. PMID- 12560916 TI - Cholecystokinin and cholecystokinin receptors. PMID- 12560917 TI - Aberrant expression of CDX2 in Barrett's epithelium and inflammatory esophageal mucosa. AB - BACKGROUND: There have been no detailed reports directly comparing the expression of CDX1 with that of CDX2 in the inflammatory esophageal mucosa and Barrett's epithelium. The present study was designed to examine the expression of CDX 1/2 in inflammatory esophageal mucosa with or without Barrett's epithelium. METHODS: The expression of CDX1/2 genes was analyzed using the reverse transcriptase polymerase chain reaction (RT-PCR) in 34 human esophageal biopsy specimens, and CDX2 expression was also evaluated immunohistochemically, using anti-human CDX2 monoclonal antibody. The biopsy specimens for RNA extraction were taken endoscopically from esophageal mucosa with mucosal break due to gastroesophageal reflux disease (GERD), Barrett's epithelium, and normal epithelium. The expressions of mucin markers (MUC2) and intestine-specific genes (sucrase-isomal tase, human defensin-5, alkaline phosphatase) were also comparatively analyzed. RESULTS: CDX1/2 expression was not found in the normal esophageal mucosa. The prevalence of CDX1/2 mRNA expression was significantly higher in the mucosa with Barrett's epithelium than in the mucosa without Barrett's epithelium. It is noteworthy, however, that the CDX2 mRNA expression was initiated at the stage of esophagitis, when neither CDX1 nor intestine-specific genes had emerged yet. In contrast to CDX2, CDX1 was expressed only in Barrett's epithelium. Immunohistochemical study demonstrated strong and extensive nuclear immunoreactivity for CDX2 in Barrett's epithelium. Furthermore, fine granular cytoplasmic staining was also observed in the cytoplasm in Barrett's epithelium, as well as in inflammatory esophageal mucosa. CONCLUSIONS: We report here, for the first time, that CDX2 is expressed in patients with Barrett's epithelium and inflammatory esophageal mucosa. These findings imply that the expression of CDX2 may be an early event leading to the development of Barrett's esophagus. PMID- 12560918 TI - Prevalence of Barrett's esophagus in an urban Korean population: a multicenter study. AB - BACKGROUND: It had been believed that over 75% of cases of esophageal carcinoma were squamous cell carcinoma (SCC). However, recently, in Western countries, the incidence of SCC has decreased, while the rate of adenocarcinoma has increased dramatically, accounting for almost 50% of esophageal carcinomas. With this change, concern over Barrett's esophagus (BE) has increased. Traditionally, it was believed that BE affected Asians less frequently. However, accelerated westernization has changed the lifestyle of Asians dramatically, and this raises the question that the prevalence might be changed in Asians living in Asia. Lacking reports on the prevalence of BE in the Korean population, we examined the data from endoscopic examinations performed at five referral hospitals in Seoul, Korea, to estimate the period prevalence of BE. METHODS: One thousand five hundred and fifty-three patients who presented themselves to five referral hospitals due to various gastrointestinal symptoms prospectively underwent upper gastrointestinal endoscopic examinations and were enrolled in the study. Two biopsy specimens were taken from any parts of pink mucosa 3 cm or more above the squamocolumnar junction. RESULTS: BE was detected in 5 of the 1553 patients who underwent upper gastrointestinal endoscopic examinations, and the approximate period prevalence of BE in Korean patients in the five referral hospitals was 3.2 per 1000. Of the 5 patients, two were men and three women with a mean age of 65 +/- 11.8 (mean +/- SD) years. All these patients had a body mass index (BMI) of less than 25 and were not obese. Regurgitation was described by 2 (40%) patients with BE, and heartburn was complained of by 4 (80%) patients. Of the 5 patients with BE, 3 (60%) patients had esophagitis, 2 (40%) patients had hiatal hernia, and none of the patients had Helicobacter pylori infection. The gross type of BE consisted of type I (spike) in 2 (40%) patients, type II (circumferential type II) in 1 (20%) patient, type III (circumferential type III) in 1 (20%) patient, and mixed type in 1 (20%) patient. Specialized intestinal metaplasia was found in all 5 patients. CONCLUSIONS: The period prevalence of BE among the patients of five referral hospitals was 3.2 per 1000, and it seems that the prevalence of BE in Korean urban area is lower than that in Western countries. PMID- 12560919 TI - Low presence of p53 abnormalities in H pylori-infected gastric mucosa and in gastric adenocarcinoma. AB - BACKGROUND: Alterations of the p53 gene and/or its abnormal protein accumulation have been observed in gastric cancer and preneoplastic lesions. Our aim was to assess possible associations between different H. pylori strains and p53 abnormalities in patients with dyspepsia and with gastric cancer. METHODS: Seventy-five dyspeptic patients and 40 patients with gastric adenocarcinoma entered the study. H. pylori status was determined by the rapid urease test, histology, and polymerase chain reaction (PCR) analysis. Overexpression of the p53 protein was evaluated by immunohistochemistry. Detection of p53 mutations was done by direct DNA sequencing. RESULTS: Fifty-four of the 75 (72.0%) dyspeptic patients and 27 of the 40 (67.5%) gastric cancer patients showed H. pylori infection. Cytotoxin-associated gene ( cagA)-positive strains were found in 31 of the 54 (58%) dyspeptic patients and in 25 of the 27 (92.6%) neoplastic patients. As regards vacA, s2 strains showed the highest prevalence among dyspeptic patients (24 of 54 patients; 44.4%), whereas s1 strains were more expressed among cancer patients (23 of 27; 85.2%). Among the dyspeptic patients, 1 patient with duodenal ulcer showed p53 overexpression. Three mutations were identified by DNA sequencing: one in a patient with normal endoscopic findings and two in patients suffering from gastritis. Among the neoplastic patients, 16 subjects (40%) showed p53 overexpression (9 had diffuse-type and 7 intestinal-type cancer). Four mutations (10%) occurred in patients with intestinal-type gastric cancer. No association between p53 abnormalities (overexpression/mutation) and H. pylori infection was found in either group of patients. CONCLUSIONS: These results lead us to hypothesize that H. pylori infection does not affect the p53 pattern in gastric mucosa. Moreover, mutations of the p53 gene do not seem to be a predominant event in gastric carcinogenesis, at least in our populations. PMID- 12560920 TI - Clinical utility of grading criteria for submucosal invasion in the prognosis of T1 colorectal carcinomas. AB - BACKGROUND: The clinical utility of relative and absolute grading criteria for submucosal invasion in T1 colorectal carcinomas has been controversial. METHODS: In 51 T1 colorectal carcinomas, depth of submucosal invasion was graded either according to a modified Haggitt's classification (a relative criterion) or by direct measurement using a micrometer (an absolute criterion), and immunostaining for E-cadherin, alpha-catenin, beta-catenin, matrilysin, and CD44 variant 6 was performed on formalin-fixed, paraffin-embedded sections. The associations between lymph node metastasis or local recurrence (locoregional failure) and tumor budding, and clinicopathologic parameters and immunoreactivity were examined statistically. RESULTS: By univariate analysis, tumor budding, histology, and the co-expression pattern of nuclear beta-catenin and CD44 variant 6 were significantly associated with locoregional failure. The relative and absolute grading of submucosal invasion were not significantly associated with locoregional failure. Multivariate analysis showed that tumor budding alone was significantly associated with locoregional failure, and the association between the co-expression pattern of nuclear beta-catenin and CD44 variant 6, and locoregional failure was marginally significant ( P = 0.0502). Lymphatic invasion and absolute grading of depth and width of submucosal invasion were significantly associated with tumor budding, and the associations between tumor budding, and histologic differentiation and membranous alpha-catenin expression were marginally significant ( P = 0.06; P= 0.08), whereas, a relative grading of submucosal invasion was not significant ( P = 0.58). Analysis of variance showed that histologic differentiation and lymphatic invasion were independently and significantly associated with tumor budding ( P = 0.005; P < 0.001). CONCLUSIONS: These results suggest that the grading of submucosal invasion, either relative or absolute, may not be a useful risk factor for lymph node metastasis or local recurrence in T1 colorectal carcinomas. PMID- 12560921 TI - Diagnostic value of the copper/zinc ratio in hepatocellular carcinoma: a case control study. AB - BACKGROUND: The aim of this study was to assess the accuracy of the copper/zinc ratio in the evaluation of a group of patients with hepatocellular carcinoma (HCC). METHODS: A total of 105 patients were studied and separated into three groups: group I ( n = 40), patients with HCC, group II ( n = 25), patients with liver cirrhosis, and group III ( n = 40), patients with benign digestive disease. Serum levels of copper and zinc were measured by atomic absorption spectrophotometry. RESULTS: The serum levels of copper microg/dl) in patients with HCC (97.4 +/- 27.2; P < 0.05) were significantly higher than those in patients with liver cirrhosis (73.7 +/- 17.5) or benign digestive disease (77.1 +/- 20.8), and the serum levels of zinc microg/dl) were significantly lower (71.6 +/- 30.5; P < 0.05) than those in patients with benign digestive disease (81.7 +/ 17.7 microg/dl) and were similar to those in cirrhotic patients (68.5 +/- 17.1). The Cu/Zn ratio was also significantly higher in patients with HCC (1.52 +/- 0.64; P < 0.05) than in patients with liver cirrhosis (1.06 +/- 0.2) or patients with benign digestive disease (0.95 +/- 0.39). Considering a cutoff value of 1.15, the sensitivity of the Cu/Zn ratio was 87.5%, with a specificity of 86.1%, a positive predictive value of 79.5%, and a negative predictive value of 91.8%. CONCLUSIONS: The Cu/Zn ratio was found to be significantly higher in patients with HCC compared with that in age and sex-matched controls, with a sensitivity of 87.5%; this ratio might be useful in the evaluation of suspected hepatocellular malignancy. PMID- 12560922 TI - A scoring system for primary biliary cirrhosis and its application for variant forms of autoimmune liver disease. AB - BACKGROUND: Although primary biliary cirrhosis (PBC) and autoimmune hepatitis (AIH) are two independent autoimmune liver diseases, it is sometimes difficult to characterize the variant forms of autoimmune liver disease. A PBC scoring system, in combination with the AIH scoring system may be helpful to characterize such patients. METHODS: A PBC scoring system was introduced that selected 14 categories characteristic of PBC. One hundred and thirty-four patients with PBC, 31 patients with autoimmune cholangitis (AIC), 22 patients with overlap syndrome, and 48 patients with AIH were included in the study. The AIC patients fulfilled the PBC criteria but were negative for anti-mitochondrial antibody and positive for anti-nuclear antibody. Overlap syndrome patients fulfilled both the PBC and AIH criteria. RESULTS: The total scores (means +/- SD) for the PBC, AIC, overlap syndrome, and AIH patients were 23.3 +/- 4.7, 9.3 +/- 4.4, 18.0 +/- 5.9, and 3.6 +/- 3.3, respectively. When definite and probable PBC patients were defined as those with a total score of over 17 and 9-17, respectively, all except for 1 patient could be classified as definite or probable PBC. Four of the 48 AIH patients were classified as probable PBC. PBC scores for the variant autoimmune liver diseases showed a wide deviation. Plotting both PBC and AIH scores in a rectangular coordinate enabled us to locate each patient with variant forms according to the deviation from classical PBC or AIH. CONCLUSIONS: The PBC scoring system might be useful in characterizing the features of variant forms of autoimmune liver disease. PMID- 12560923 TI - Influence of albumin binding on the substrate transport mediated by human hepatocyte transporters OATP2 and OATP8. AB - BACKGROUND: Despite their strong binding to albumin while circulating in blood, many organic anions, such as bilirubin and fatty acids, are removed efficiently by the liver. The uptake transporters of human hepatocytes, OATP2 (symbol, SLC21A6) and OATP8 (SLC21A8), play important roles in the hepatic uptake of endogenous substances and drugs. The two transporters show different affinities for the organic anion sulfobromophthalein (BSP), which binds with high affinity to albumin in blood. METHODS: In this study, we investigated whether a direct interaction of albumin with OATP2 or OATP8 occurs during the uptake of BSP. The uptake of BSP, at varying concentrations of human serum albumin (HSA), into transfected HEK293 cells expressing recombinant human OATP2 or OATP8 was measured. The influence of other organic anions on the uptake of albumin-bound BSP by OATP2 or OATP8 was also studied. RESULTS: OATP8-mediated transport was affected more strongly by HSA than OATP2-mediated transport. Albumin affected both transporters in the manner of a noncompetitive inhibitor. Uptake studies using OATP2-transfected MDCKII cells indicated that a direct interaction between albumin and OATP2 is not necessary for uptake, a finding that was further confirmed by the effects of bilirubin and palmitate on the binding of BSP to albumin and on the uptake of BSP by OATP2 or OATP8. CONCLUSIONS: Our results indicated that uptake of albumin-bound BSP occurs only from the pool of unbound ligand. PMID- 12560924 TI - Increased cyclooxygenase-2 expression in large flat colorectal tumors (laterally spreading tumors). AB - BACKGROUND: Little is known about the molecular biological features of large flat tumors or laterally spreading tumors (LSTs) of the colon and rectum. METHODS: The cyclooxygenase (COX)-2 protein expression level of LSTs was assessed immunohistochemically and compared with that of exophytic colorectal tumors. COX 2 expression in the dysplastic epithelial cells and the interstitial cells was evaluated independently. RESULTS: Sixty-five LSTs and 65 exophytic tumors were randomly selected from the archives, and 61 and 59, respectively, were successfully analyzed. LSTs showed significantly more intense COX-2 protein expression in the dysplastic epithelial cells than the exophytic tumors ( P > 0.0001). COX-2-positive interstitial cells were also observed more frequently in LSTs than in the exophytic tumors ( P = 0.015). CONCLUSIONS: Overexpression of COX-2 protein may play a crucial role in the tumorigenesis of colorectal LSTs. Chemoprevention, using nonsteroidal anti-inflammatory drugs (NSAIDs) or selective COX-2 inhibitors may be effective for treating this unique neoplasm, which sometimes defies endoscopic treatment. PMID- 12560925 TI - A case of primary esophageal tuberculosis diagnosed by identification of Mycobacteria in paraffin-embedded esophageal biopsy specimens by polymerase chain reaction. AB - We present a case of primary esophageal tuberculosis diagnosed by polymerase chain reaction (PCR) of paraffin-embedded esophageal biopsy specimens. A 42-year old Japanese woman visited our clinic because of dysphagia. Radiologic and endoscopic examinations revealed a stenotic lesion with reddish mucosa and multiple ulcers in the middle esophagus. There was no associated lesion outside the esophagus. Histological and bacteriological studies of esophageal biopsy specimens and gastric aspirates did not give a definitive diagnosis. However, mycobacterial DNA was detected by PCR of paraffin-embedded esophageal biopsy specimens. She then was diagnosed as having primary esophageal tuberculosis. The esophageal mucosal lesion almost healed after 1 month of antituberculosis medication with residual annular stenosis which was resolved later by endoscopic balloon dilation. PMID- 12560926 TI - Association of hereditary spherocytosis with familial adenomatous polyposis in a pedigree: a new syndrome or coincidence? AB - No association of familial adenomatous polyposis (FAP) and hereditary spherocytosis (HS) has been reported, both of which are inherited in an autosomal dominant manner. We present the first reported case of FAP with spherocytosis and construct the family pedigree. In the patient's pedigree, both FAP and spherocytosis were inherited in an autosomal dominant trait. In the 34-year-old Japanese proband's leukocytes, we found no abnormal chromosomal band, and a germline mutation of the APC gene was not detected. All possible genes reported to be linked to HS were located far from chromosome 5q on which the APC gene is located. Although it is unknown if erythrocyte membrane disorder is an additional phenotype of FAP, to the best of our knowledge, this is the first documentation of FAP associated with spherocytosis. PMID- 12560927 TI - Ultrasonographic findings of amebic colitis. AB - We present here a case of a 64-year-old man with amebic colitis who was examined by ultrasonography. Gray-scale ultrasonography revealed marked thickening of the bowel wall, specifically thickening of the submucosal layer. Color Doppler ultrasonography showed hypervascularity of the submucosal and proper muscle layers. With effective treatment, thickening and hypervascularity of the bowel wall disappeared promptly. This is the first report demonstrating ultrasonographic findings of amebic colitis. We conclude that information provided by ultrasonography is useful not only for detecting the bowel abnormality of amebic colitis but also for evaluating the therapeutic effect on amebic colitis. PMID- 12560928 TI - Novel germline mutation of the PTEN gene in a Japanese family with Cowden disease. AB - We report a novel germline mutation of the PTEN gene in a Japanese family with Cowden disease. A 46-year-old Japanese man and his mother were diagnosed as having Cowden disease. Their physical examinations revealed multiple facial trichilemmoma, oral mucosal papillomatosis, palmoplantar keratosis, and gastrointestinal polyposis. The single-strand conformation polymorphism (SSCP) analysis showed an abnormal band on exon 7 of their PTEN gene. Direct sequence analysis of exon 7 detected a TAAA insertion to codon 221, producing a stop codon (c.663ins TAAA). PMID- 12560929 TI - Fulminant amoebic colitis with perforation successfully treated by staged surgery: a case report. AB - Radical surgery for fulminant amoebic colitis leads to extremely high mortality; however, resective surgery is mandatory if a patient develops massive fecal peritonitis. We herein report an extremely rare case of fulminant amoebic colitis with multiple perforations, which was successfully treated by staged surgical procedures. A 48-year-old man who had been treated with predonisolone under a diagnosis of ulcerative colitis was admitted. Biopsy specimens from the colonic mucosa revealed Entamoeba histolytica. On the day of diagnosis, he developed severe abdominal pain and underwent emergency laparoptomy, showing total colonic gangrene with multiple perforations associated with massive fecal peritonitis. Subtotal colectomy, mucous fistula of the rectosigmoid, and ileostomy were performed. He recovered well although disseminated intravascular coagulopathy developed postoperatively. As the middle and upper part of rectum was found to be severely stenotic 4 months after surgery, we performed proctectomy, ileal pouch anal canal anastomosis, and diverting ileostomy, which was reversed 6 months later. The patient has been well with satisfactory anal function 37 months after the initial surgery. This case suggests that (1). early and accurate diagnosis of amoebiasis is important to avoid surgical intervention, and (2). staged surgery including total colectomy should be considered as one of the treatment choices even in patients with total necrotizing amoebic colitis. PMID- 12560930 TI - A case of triple synchronous cancers occurring in the gallbladder, common bile duct, and pancreas. AB - We report a 74-year-old man with triple synchronous cancers occurring in the gallbladder, common bile duct, and pancreas. The patient had consulted a nearby physician because of epigastralgia and icterus. On September 30, 1997, the patient was admitted to our department for further evaluation and treatment. Abdominal computed tomography (CT) showed dilatation of the common bile duct, cystic duct, and intrahepatic bile duct, and swelling of the gallbladder. On CT, the wall of the distal common bile duct was thick and a low-density mass was detected on the left side. Cholangiography, performed via percutaneous transhepatic cholangiodrainage (PTCD), revealed stenosis of the distal common bile duct. Endoscopic retrograde pancreatography (ERP) showed marked dilatation of the main pancreatic duct. On October 17, 1997, pancreatoduodenectomy was performed under the diagnosis of carcinoma of common bile duct and pancreas. Histopathological examination revealed poorly differentiated tubular adenocarcinoma of the common bile duct, well-differentiated tubular adenocarcinoma of the gallbladder, and mucinous cystadenocarcinoma of the pancreas. These three tumors were histopathologically different. Moreover, p53 positive nuclei were recognized only in the pancreas tumor. These findings suggested that the oncogenic mechanisms of multiple synchronous cancers were not the result of only abnormal DNA reparative mechanisms. PMID- 12560931 TI - Transient portal vein thrombosis caused by radiofrequency ablation for hepatocellular carcinoma. PMID- 12560932 TI - The copper/zinc ratio in patients with hepatocellular carcinoma. PMID- 12560933 TI - Evaluation of a new scoring system for primary biliary cirrhosis and its related variant forms. PMID- 12560934 TI - The 'cancer cachectic factor'. AB - The object of this study was to summarize information on catabolic factors produced by tumours which lead to tissue catabolism in cancer cachexia and to use this information for the development of effective therapy. The study population was made up of patients with cancer cachexia and weight loss greater than 1 kg month(-1). They had a varied range of carcinomas, particularly pancreatic, but also of the breast, ovary, lung, colon and rectum. Cachectic factors were isolated by standard biochemical methods, and the mechanism of tissue catabolism was evaluated in vitro and in vivo. We isolated a 24-kDa sulphated glycoprotein produced by cachexia-inducing murine and human tumours, which induces catabolism of myofibrillar proteins in skeletal muscle and for this reason has been named proteolysis-inducing factor (PIF). PIF was shown to be present in a diverse range of carcinomas in patients whose rate of weight loss exceeded 1.0 kg month(-1). Administration of PIF to normal mice produced a rapid decrease in body weight, which arose primarily from a loss of skeletal muscle, accompanied by increased mRNA levels for ubiquitin, the ubiquitin-carrier protein (E2(14k)), and proteasome subunits. This suggests that PIF induces protein catabolism through an increased expression of the key components of the ATP-ubiquitin-dependent proteolytic pathway. The action of PIF was attenuated both in vitro and in vivo by eicosapentaenoic acid (EPA). Oral EPA has been found to stabilize the body weight of patients with advanced pancreatic cancer and, when combined with an energy- and protein-rich nutritional supplement, to produce weight gain arising solely from an increase in lean body mass. Nutritional supplementation alone is unable to reverse the process of muscle wasting in cancer patients, since this arises from activation of the ubiquitin proteasome pathway by PIF, which is independent of nutrient intake. EPA is able to down-regulate the increased expression of this pathway and prevents muscle wasting in cancer patients. PMID- 12560935 TI - Treatment of cancer-related fatigue: an update. AB - Fatigue, a common complaint of cancer patients, requires a multidisciplinary evaluation and treatment approach because of the multiple etiologies and contributing factors. Current treatments for fatigue include educating patients and caregivers about fatigue, applying etiology-specific treatments, utilizing nonpharmacologic interventions, and prescribing pharmacologic therapies. Often, an individualized treatment plan that includes several modalities may be developed. Presently, there is a lack of well-designed clinical trials to evaluate pharmacologic agents for the treatment of cancer-related fatigue. PMID- 12560936 TI - Normal-release and controlled-release oxycodone: pharmacokinetics, pharmacodynamics, and controversy. AB - Oxycodone has become one of the most popular opioids in the United States. It is superior to morphine in oral absorption and bioavailability, and similar in terms of protein binding and lipophilicity. Gender more than age influences oxycodone elimination. Unlike morphine, oxycodone is metabolized by the cytochrome isoenzyme CYP2D6, which is severely impaired by liver dysfunction. Controlled release (CR) oxycodone has become one of the most frequently utilized sustained release opioids in the United States. Both its analgesic benefits and its side effects are similar to those of CR morphine. CR oxycodone is similar to morphine and other opioids in its abuse potential. Deaths attributable to oxycodone are usually associated with polysubstance abuse in which oxycodone is combined with psychostimulants, other opioids, benzodiazepines or alcohol. Oxycodone's kappa receptor binding has little role in abuse or addiction. The cost of CR oxycodone is prohibitive for most American hospices. PMID- 12560937 TI - Granisetron: relating pharmacology to clinical efficacy. AB - The 5-HT(3)-receptor antagonists, which are regarded as the 'antiemetic gold standard', are used for prophylaxis against chemotherapy- and radiotherapy induced emesis. At equally effective doses, the 5-HT(3)-receptor antagonists have been shown to demonstrate broadly equivalent clinical antiemetic activity and safety. However, the pharmacology of granisetron suggests that it may have an improved side-effect and tolerability profile, a lower risk of drug interactions and a longer duration of action than other 5-HT(3)-receptor antagonists. The high affinity of granisetron, coupled with the insurmountable antagonism displayed at 5-HT(3) receptors, is thought to underlie its good clinical efficacy. PMID- 12560938 TI - Granulocyte transfusions from G-CSF-stimulated donors for the treatment of severe infections in neutropenic pediatric patients with onco-hematological diseases. AB - From March 1994 to January 2001, 15 courses of granulocyte transfusion (GTX) were administered to 13 neutropenic patients (6 male and 7 female patients; median age 7 years, range 3 months to 14 years) affected by: acute lymphoblastic leukemia (ALL) in 6 cases, acute myeloid leukemia (AML) in 5, very severe aplastic anemia in 1, and familial erythrophagocytic lymphohistiocytosis (FEL) in 1. Infections were classified as microbiologically defined and clinically defined infections in 8 and 7 episodes, respectively. Before the GTX transfusions, broad-spectrum antibacterial and antifungal therapy had been administered for a median of 12 (range 5-28) and 8 days (range 2-50), respectively, with no improvement. G-CSF was administered prior to GTX in 9 episodes of infection, with a median of 9 days of treatment (range 4-30). Leukapheresis was obtained from 15 related donors (father, 10; mother, 3; sister, 1; aunt, 1) after s.c. stimulation with G-CSF, 300 micro g daily, starting from day -3 (where day 0 was the day of the first granulocyte collection) and continuing throughout the period of GTX treatment. The donors' median white blood cell (WBC) count at leukapheresis was 31.6 x 10(9)/l (range 12-56), and the median yield was 31.39 x 10(9) WBC (range 2.96 64.73 x 10(9)), with a proportion of PMN of 90-95%. Overall, 70 GTX were administered, with a median of 4 GTX per episode of infection (range 2-11). The combination of GTX with antimicrobial therapy led to complete or partial recovery in 6 and in 3 of 15 episodes (60%), respectively. Priming of the donor with G-CSF was well tolerated, the most common side-effects being bone pain, malaise and paresthesia. All donors are alive and well after a median of 4.5 years (range 0.8 7.7) from donation. We conclude that GTX is potentially useful when the severity of the infection and the host's immunodeficiency make any other antimicrobial treatment ineffectual. Long-term safety data on the stimulation of donors with G CSF have been reassuring to date. Further controlled studies are needed to assess the exact role of GTX in the outcome of neutropenic patients with severe infection and any criteria for patient selection and the timing of GTX administration. PMID- 12560939 TI - Compatibility of haloperidol and hyoscine-N-butyl bromide in mixtures for subcutaneous infusion to cancer patients in palliative care. AB - The administration of drugs by s.c. infusion is routinely practiced in palliative medicine for the management of patients who are no longer able to take oral medication. It is not uncommon for two or more drugs to be combined in s.c. infusion solutions. Unfortunately, the compatibility and stability of haloperidol and hyoscine- N-butyl bromide has not yet been determined. The objective of this study was to study the compatibility and stability of solutions containing both drugs in polypropylene syringes. Nine different solutions were assessed for up to 15 days following preparation. The solutions were prepared in polypropylene syringes with 0.9% saline as a diluent and stored at 4 degrees C and 25 degrees C. High-performance liquid chromatography was the analytical technique used to measure haloperidol and hyoscine- N-butyl bromide. The initial concentration ranges were 0.3125-1.25 mg/ml for haloperidol and 2.5-10.0 mg/ml for hyoscine- N butyl bromide. Haloperidol was precipitated at a concentration of >/=1.25 mg/ml when it was combined with hyoscine- N-butyl bromide. Concentrations of hyoscine- N-butyl bromide lower than 10 mg/ml in mixtures with haloperidol or 0.625 mg/ml of haloperidol in mixtures with hyoscine- N-butyl bromide for s.c. infusion allow for the administration of both drugs without any significant loss after storage at 25 degrees C for periods of up to 15 days, with approximately >/=90% and 88%, respectively, of haloperidol and hyoscine- N-butyl bromide remaining. However, after storage of the mixtures for equivalent periods at 4 degrees C the losses of hyoscine- N-butyl bromide observed at the end of the study were higher than 20%, while the percentages of haloperidol remaining after 15 days at this temperature were >/=94.37%. PMID- 12560940 TI - A model of acute symptom control unit: Pain Relief and Palliative Care Unit of La Maddalena Cancer Center. AB - Palliative care in Italy was provided solely on a home care basis until a couple of years ago. In recent years different realities have been created according to personal experience, quite apart from new resources provided by the National Health System. The first Pain Relief and Palliative Care Unit in Sicily, the largest region in Italy, was established in March 1999. Most members of the regular staff of this Unit have a background in anesthesiology. Activity in the Unit has grown progressively, with 460 admissions in the last year. The characteristics of the first 1,000 patients admitted, the principal protocols for diagnostic and therapeutic procedures, such as those for hitherto intractable pain, nutrition and hydration, interventional procedures for symptom control, and emerging problems, a project called "Island with no pain," and activities in the fields of formation and research are described. PMID- 12560941 TI - Aerobic exercise for a patient suffering from metastatic bone disease. AB - A 48-year-old female patient suffering from advanced breast cancer with metastatic bone disease participated in an aerobic exercise program consisting of ergometer cycling three times a week for 1 year. Feasibility, safety and beneficial effects of the program were proven for the patient in this case study. VO(2)max (20.2% after 16 weeks, 52.7% after 12 months) and peak work capacity (15.5% after 16 weeks, 35.7% after 12 months) had increased. The patient experienced a marked improvement in physical performance and in quality of life. Aerobic exercise, initiated and executed with appropriate care, may serve as a useful additional means of palliative treatment in some cancer patients with bone metastases. PMID- 12560942 TI - Post-traumatic stress disorder associated with suspected lung cancer and bereavement: 4-year follow-up and review of the literature. AB - We report here a patient who developed post-traumatic stress disorder (PTSD) after being given a diagnosis of suspected lung cancer. The symptoms of PTSD continued even after it had been confirmed that the lesion was benign after all. A 73-year-old man was referred to our psychiatric outpatient clinic for depressed mood, appetite loss and difficulty in sleeping. On examination the patient explained that he had become preoccupied with intrusive thoughts and memories since his diagnosis of suspected lung cancer and that although he had been told the lesion was not malignant, he had distressing memories of that period and was experiencing severe psychological distress. The patient also explained that he had experienced the death of a close friend through lung cancer and he had a deep fear of developing lung cancer himself. In this patient, the psychological distress evoked by the diagnosis of suspected cancer was severe enough to induce the symptoms of PTSD even though the tumor was benign. From the clinical point of view, it could provide useful information if liaison psychiatrists were to ask patients about any experience of losing significant others to the same disease and this might be helpful in gaining an understanding of the disease process. PMID- 12560943 TI - Successful treatment of intractable hiccup with methylphenidate in a lung cancer patient. AB - In a 56-year-old man with metastatic small-cell lung cancer, a persistent hiccup was refractory to classic treatments. Hyponatremia, neoplasic mediastinal involvement and liver metastases were though to be potential causative factors. Methylphenidate (MTP), a mild central nervous system (CNS) stimulant that is most commonly used for the treatment of attention deficit hyperactivity disorder, was started at a daily dose of 10 mg. It was rapidly efficient and well tolerated. This report suggests a potential advantage of MTP in the treatment of intractable hiccup in cancer patients. PMID- 12560944 TI - GJB2 deafness gene shows a specific spectrum of mutations in Japan, including a frequent founder mutation. AB - Mutations in the GJB2 gene (connexin 26) are the major cause of autosomal recessive non-syndromic hearing impairment in many populations. In contrast to the volume of information regarding the involvement of GJB2 mutations in hearing impairment in populations of European ancestry, less is known regarding other ethnic groups. In this study, we analyzed the GJB2 gene for mutations in 1227 hearing-impaired Japanese individuals. This revealed a unique spectrum of GJB2 mutations, different from that found in the Caucasian population. The most frequent mutation in Japanese, 235delC, has never been reported in Caucasians. To investigate a possible founder effect for the 235delC mutation, we analyzed single nucleotide polymorphisms in the vicinity of the GJB2 gene. Results were consistent with inheritance of the 235delC mutation from a common ancestor. The results of this study have important implications for genetic diagnostic testing for deafness in the Japanese population. PMID- 12560945 TI - The signature motif in human glucose-6-phosphate transporter is essential for microsomal transport of glucose-6-phosphate. AB - Glycogen storage disease type Ib (GSD-Ib) is caused by a deficiency in the glucose-6-phosphate transporter (G6PT). Sequence alignments identify a signature motif shared by G6PT and a family of transporters of phosphorylated metabolites. Two null signature motif mutations have been identified in the G6PT gene of GSD Ib patients. In this study, we characterize the activity of seven additional mutants within the motif. Five mutants lack microsomal G6P uptake activity and one retains residual activity, suggesting that in G6PT the signature motif is a functional element required for microsomal glucose-6-phosphate transport. PMID- 12560947 TI - Austrian Moderate Altitude Study (AMAS 2000) - fluid shifts, erythropoiesis, and angiogenesis in patients with metabolic syndrome at moderate altitude (congruent with 1700 m). AB - It was hypothesized that subjects with metabolic syndrome (hypertension, obesity, hyperlipidemia, diabetes mellitus): (1) develop measurable peripheral edema at moderate altitude and (2) might show differences on erythropoiesis, iron status and vascular endothelial growth factor (VEGF) in comparison to healthy subjects during and after a long-term stay (3-week exposure) at moderate altitude (congruent with 1700 m). Twenty-two male subjects with metabolic syndrome were selected. Baseline investigations (t1) were performed in Innsbruck (500 m). All participants were transferred by bus to 1700 m (Alps) and remained there for 3 weeks with examinations on day 1 (after the first night at altitude, t2), day 4 (t3), day 9 (t4) and day 19 (t5). After returning to Innsbruck, post-altitude examinations were conducted after 7-10 days (t6) and 6-7 weeks (t7), respectively. Body mass was decreased from t1 to t7 (P<0.01). Total body water was decreased at t2 (P<0.01), returned to control level (t3, t4), and was found elevated at t7 (P<0.01). Lean body mass did not change, but body fat decreased during the study (P<0.01). Tissue thickness at the forehead decreased during and after altitude exposure (P<0.01), whereas tissue thickness at the tibia did not alter. Erythropoietin (EPO) was elevated as early as t2 and remained increased until t5. Reticulocyte count was increased at t3 and remained above pre-altitude values. VEGF levels were unchanged. After a 3-week exposure to moderate altitude, patients with metabolic syndrome had reduced their body mass, mainly because of a reduction in body fat. The moderate altitude was found to stimulate erythropoiesis in these patients but this was not sufficient to increase serum VEGF concentration. PMID- 12560946 TI - Relationship between in vivo muscle force at different speeds of isokinetic movements and myosin isoform expression in men and women. AB - In an attempt to explore the relationship between force production during voluntary contractions at different speeds of isokinetic movement and the myofibrillar protein isoform expression in humans, an improved isokinetic dynamometer that corrects for gravitation, controls for acceleration and deceleration, and identifies a maximum voluntary activation was used. Muscle torque recordings were compared at the same muscle length (knee angle) and the torque was calculated as the average torque at each angle over a large knee angle range (75 degrees -25 degrees ) to reduce the influence of small torque oscillation on the calculated torque. Muscle torque at fast (240 degrees s(-1)) versus slow (30 degrees s(-1)) speeds of movement, torque normalized to muscle cross-sectional area (specific tension), and absolute torque at fast speeds of movement were measured in 34 young healthy male and female short-, middle-, and long-distance runners. The relationship between the different measures of muscle function and the expression of myosin heavy chain (MyHC) isoforms using enzyme histochemical and electrophoretic protein separation techniques were investigated. A significant correlation between the 240 degrees s(-1) vs 30 degrees s(-1) torque ratio and the relative area of the type II fibers and type II MyHC isoforms were observed in both the men (r=0.74; P<0.001) and the women (r=0.81; P<0.05). Thus, the present results confirm a significant relationship between in vivo human muscle function and the MyHC isoform expression in the contracting muscle. PMID- 12560948 TI - Austrian Moderate Altitude Study 2000 (AMAS 2000). The effects of moderate altitude (1,700 m) on cardiovascular and metabolic variables in patients with metabolic syndrome. AB - We investigated the changes in the cardiovascular system [resting blood pressure (BP) and heart rate (HR), measured by means of a 24-h ambulatory BP and a holter electrocardiogram (ECG)], glycemic parameters, and lipid metabolism of subjects suffering from metabolic syndrome during a 3-week sojourn at 1,700 m in the Austrian Alps. A total of 22 male subjects with metabolic syndrome were selected. Baseline investigations were performed at Innsbruck (500 m above sea level). During the 3-week altitude stay the participants simulated a holiday with moderate sports activities. Examinations were performed on days 1, 4, 9, and 19. After returning to Innsbruck, post-altitude examinations were conducted after 7 10 days and 6-7 weeks, respectively. The 24-h ambulatory BP and holter ECG revealed a decrease in average HR, BP, and rate pressure product (RPP: systolic blood pressure x HR) after 3 weeks of altitude exposure. In some patients, an increase in premature ventricular beats was observed at the end compared to the beginning of the exposure to moderate altitude. The ECG revealed no ischemic ST segment changes. Maximal physical capacity as measured by symptom-limited maximal cycle ergometry tests remained unchanged during the study. Six weeks after the altitude exposure the blood pressure increased again and returned to pretest levels. The Homeostasis Model Assessment index, which is a measure of insulin resistance, decreased significantly and glucose concentrations obtained after an oral glucose tolerance test were significantly lower after the stay at altitude compared to the basal values. We conclude that after a 3-week exposure to moderate altitude, patients with metabolic syndrome (1) tolerated their sojourn without any physical problems, (2) exhibited short-term favorable effects on the cardiovascular system, and (3) had significant improvements in glycemic parameters that were paralleled by a significant increase in high-density lipoprotein-cholesterol. PMID- 12560949 TI - Dynamic changes of deleted mitochondrial DNA in human leucocytes after endurance exercise. AB - We examined the time course of the appearance and disappearance of deleted mitochondrial DNA (mtDNA) in human leucocytes after endurance exercise. Five untrained healthy females [aged 20.2 (0.4) years] exercised for 30 min at 50-60 W at 60 rpm on a cycle ergometer for 2 or 3 consecutive days. Blood samples were collected immediately before exercise, and on three occasions after the end of the last exercise bout. Levels of mtDNA were analyzed using the nested polymerase chain reaction method. Prior to exercise, a common mtDNA deletion was identified in all subjects. This common deletion was again identified in all subjects 1 day after completion of the exercise regime. However, mtDNA with this common deletion was found to have disappeared 2 days after exercise in three subjects, and after 4 days in the other two subjects. The deletion then reappeared 5 or 6 days after the final exercise bout in three of the five subjects. These findings demonstrate that a common deletion in leucocytes disappears over a period of several days after endurance exercise and reappears a number of days thereafter. Therefore, the status of the appearance and disappearance of common mtDNA deletion in leucocytes is highly dynamic. PMID- 12560950 TI - Gender differences in the viscoelastic properties of tendon structures. AB - The purpose of this study was to investigate the differences in the viscoelastic properties of human tendon structures (tendon and aponeurosis) in the medial gastrocnemius muscle between men (n=16) and women (n=13). The elongation of the tendon and aponeurosis of the medial gastrocnemius muscle was measured directly by ultrasonography, while the subjects performed ramp isometric plantar flexion up to the voluntary maximum, followed by a ramp relaxation. The relationship between the estimated muscle force (Fm) and tendon elongation (L) during the ascending phase was fitted to a linear regression, the slope of which was defined as stiffness. The percentage of the area within the Fm- L loop to the area beneath the curve during the ascending phase was calculated as hysteresis. The L values at force production levels beyond 50 N were significantly greater for women than for men. The maximum strain (100xDeltaL/initial tendon length) was significantly greater in women [9.5 (1.1)%] than in men [8.1 (1.6)%]. The stiffness and Young's modulus were significantly lower in women [16.5 (3.4) N/mm, 277 (25) MPa] than in men [25.9 (7.0) N/mm, 356 (32) MPa]. Furthermore, the hysteresis was significantly lower in women [11.1 (5.9)%] than in men [18.7 (8.5)%, P=0.048]. These results suggest that there are gender differences in the viscoelastic properties of tendon structures and that these might in part account for previously observed performance differences between the genders. PMID- 12560951 TI - Exhausting stretch-shortening cycle (SSC) exercise causes greater impairment in SSC performance than in pure concentric performance. AB - The purpose of the present study was to investigate the fatigue effect of repeated exhaustive stretch-shortening cycle (SSC) exercise on concentric muscle function. Ten healthy male subjects performed SSC exercise [92 (30) jumps] on a special sledge apparatus. Exhaustion occurred on average within 3 min. A squat jump (SJ) test utilizing a concentric-only action was performed immediately before and after the SSC exercise, and then 10 min, 20 min, 2 days and 4 days later. In addition, a drop jump (DJ) test using an SSC was also performed immediately before and 20 min after the SSC exercise, and 2 days and 4 days later. During jump tests, lower limb joint moment, power, and work contributions were analyzed by using the kinetic and kinematic data. The fatigue exercise was characterized by a relatively high blood lactate concentration [7.2 (0.8) mmol x l(-1)] and a 2-day delayed increase in serum creatine kinase activity [486 (300) U x l(-1)]. SJ performance decreased markedly immediately after the SSC exercise (P<0.05) and then recovered within 10 min. In contrast, DJ performance and knee joint contribution showed a delayed decrease 2 days after the SSC exercise bout. The surface electromyographic (EMG) activity of the lower limb muscles showed no obvious change in the SJ in comparison to the DJ, although in the latter there was a delayed decrease of knee extensor EMG during the pre-activation and braking phases. The results suggest that isolated concentric muscle function is affected mainly by acute metabolic fatigue after SSC exercise. During a follow-up period after the exercise, changes in hip and knee joint contribution in SJ showed a different recovery pattern compared to those in eccentric DJ. It could be suggested that exhaustive SSC exercise would mainly influence the relative power work balance between the hip and knee joints during the eccentric phase of SSC. Thus different motor control strategies may account for the distinctive fatigue responses observed in SJ and DJ. PMID- 12560952 TI - Long-term exposure to intermittent hypoxia results in increased hemoglobin mass, reduced plasma volume, and elevated erythropoietin plasma levels in man. AB - While it is well established that highlanders have optimized their oxygen transport system, little is known about the acclimatization of those who move between different altitudes. The purpose of this study was to establish whether the acclimatization to long-term intermittent hypoxic exposure in members of the Chilean Army who frequently move from sea level to 3,550 m altitude is correlated with acute acclimatization or chronic adaptation to hypoxia. A group of officers was exposed intermittently to hypoxia for about 22 years (OI, officers at intermittent hypoxia) and a group of soldiers for 6 months (SI, soldiers at intermittent hypoxia). Both groups were compared to residents at altitude (RA) and to soldiers at sea level (SL). When compared to SL, we observed an 11% increase in total hemoglobin mass (tHb) as well as a corresponding increase in red cell volume (RCV), hemoglobin concentration and hematocrit in all three groups at altitude. Plasma volume (PV) and blood volume (BV) decreased at altitude but increased when OI and SI returned to sea level. Moreover, intermittent hypoxic exposure of OI and SI resulted in increased plasma erythropoietin (Epo) levels, which peaked on day 2 at high altitude followed by decreasing levels during the successive days, and reaching pre-altitude values in SI even when staying at altitude. In conclusion, with regard to tHb and RCV, the acclimatization to long-term intermittent hypoxia resembles the adaptation to chronic hypoxia, while PV and BV regulation mimicked acclimatization to acute hypoxia. Remarkably, finely controlled regulation of Epo expression still occurs after up to 22 years of weekly exposure to altitude. PMID- 12560953 TI - The respiratory effects of two modes of passive exercise. AB - We monitored gas exchange and muscle activity during two commonly used modes of passive leg exercise as a means of assessing the degree of passivity associated with these techniques. Additionally, we measured the rapid changes in ventilation at the start and end of the passive exercise to assess changes that occur in the fast exercise drive to breathe during passive exercise. We monitored seven subjects at rest and during 5 min of passive exercise using (1) cycling movements performed on a tandem bicycle and (2) leg extension movements performed in a chair apparatus. The increase in measured parameters from rest to passive exercise were all higher while using the tandem bicycle compared to the chair apparatus: ventilation [3.09 (0.63) versus 0.35 (0.38) l x min(-1)], leg muscle electromyogram [8.3 (1.6) versus 1.6 (0.6) microV], carbon dioxide production [0.092 (0.018) versus -0.002 (0.001) l x min(-1)] and oxygen consumption [0.158 (0.046) versus -0.008 (0.007) l x min(-1)]. Passive exercise on the tandem bicycle was also associated with a significantly larger fast exercise drive to breathe at both the start and end of motion, compared to the chair apparatus [start: 5.59 (0.74) versus 3.08 (0.79) l x min(-1), end: 3.38 (0.79) versus 1.75 (0.54) l x min(-1)]. The fast exercise drive to breathe at the end of exercise was significantly smaller (60%) than at the start for both modes. We conclude that passive exercise on an upright bicycle contains a significant active component that contributes both neural and metabolic influences toward the physiological adjustments observed. Furthermore, we conclude that adaptation occurs in the fast exercise drive to breathe accompanying passive limb movement. PMID- 12560954 TI - Characteristics of titin in strength and power athletes. AB - The purpose of this investigation was to identify characteristics of the muscle protein titin in different athletic populations with increased levels of strength and power relative to non-athletes. Subjects fell into one of four groups: (1) non-athletes (NA) ( n=5), (2) weightlifters (WL) (n=5), (3) powerlifters (PL) (n=5), (4) sprinters (S) (n=5). A one repetition maximum in the squat exercise was performed to assess strength. In addition, countermovement vertical jump trials were performed to assess power capabilities. Peak power (W(peak)) was calculated for the vertical jumps from force plate measurements. From gel electrophoresis analyses of muscle samples, titin-1 (T1) and titin-2 (T2) protein bands were identified, quantified and expressed relative to each other. In addition the relative mobility (R(f)) of T1 and T2 was determined as an estimate of molecular weight. The NA group [%T1=47.8 (5.1), %T2=52.2 (5.1), mean (SE)] had lower T1 and higher T2 percentages than WL [%T1=62.3 (6.6), %T2=37.7 (6.6)], PL [%T1=66.8 (5.0), %T2=33.2 (5.0)] and S [%T1=65.9 (4.9), %T2=34.1 (4.9)] groups (P< or =0.10, preliminary investigation into titin and exercise justifies more liberal alpha level). No significant differences were found in R(f) of T1 or T2 between the groups. This investigation has shown that there is a differential expression of titin protein bands in competitive athletes with increased levels of strength and power in comparison to untrained non-athletic individuals. Some relationships between titin characteristics and athletic performance were observed; however, no conclusions can be made based on these data as to the contribution of titin to strength or power capabilities. PMID- 12560955 TI - Electromechanical assessment of ankle stability. AB - The goal of this study was to approach ankle instability by measuring the electromechanical delay of the peroneal muscles (foot pronators). For that purpose, supramaximal electrical stimulation of common peroneal nerve was applied when the subject was standing on a force plate in bipedal stance or monopedal stance, postures requiring greater ankle stabilization. The electromechanical delay (EMD) was defined as the time interval between the onset of the peroneus longus (PL) electromyogram detected by surface electrodes and the onset of the lateral ground reaction force (Fy) measured on a force plate. Ten healthy subjects (control group, C-G) and ten subjects with functional ankle instability ("FAI" group, FAI-G) performed the tests. In C-G, the mean EMD values decreased significantly ( P<0.001) from the bipedal [10.54 (0.71) ms; mean (SD)] to the monopedal stance [8.67 (0.63 ms)]. Since a monopedal stance is known to require higher leg muscle tone resulting in a higher ankle stiffness, these results indicate that PL EMDs are sensitive to musculo-tendinous stiffness. They validate the choice of a PL EMD measurement as an indirect index of musculo-tendinous stiffness at the ankle. In both bipedal and monopedal stance conditions, EMD values were significantly higher in FAI subjects. They ranged from 12.64 (1.14) ms in the bipedal stance to 10.85 (1.07) ms in the monopedal stance. This suggests a lower musculo-tendinous stiffness at the ankle, which may contribute to the ankle instability. PMID- 12560956 TI - Effect of eccentric exercise on patients with chronic exertional compartment syndrome. AB - There are a number of similarities between chronic exertional compartment syndrome (CECS) and the effects of high force eccentric contractions in healthy controls. We hypothesised that CECS patients would be particularly susceptible to pain, fatigue and swelling after eccentric exercise. Ten CECS patients [aged 30.3 (8.0) years, mean (SD)], 7 males) and 14 healthy controls [aged 32.3 (9.0) years, 7 males] performed 40 maximal eccentric contractions of the anterior tibial muscles at an angular velocity of 90(o) s(-1). Maximal voluntary isometric contractions (MVC), force generated by electrical stimulation at 10 Hz and 50 Hz and muscle thickness (measured by real-time ultrasound scanning) were measured before exercise and for 10 min afterwards. Ratings of pain and tenderness were made before exercise, then 24 h and 48 h later. The two groups were comparable for initial isometric strength and muscle size. MVC force immediately after exercise fell to 90 (2.5)% [mean (SEM)] in patients and 86.5 (2.4)% in controls (P<0.0005 for both groups) and had not recovered after 10 min. Preferential loss of force at the low stimulation frequency occurred in both groups to a similar extent (20%, P<0.0005) and continued to decline over 10 min of recovery. There was no significant change in muscle thickness in either group. Only mild pain and tenderness were reported by the controls. The patients showed great individual variation but their mean pain scores were significantly higher during dorsiflexion and palpation (P<0.05) 24 h after exercise. After 48 h the patients reported more pain during dorsiflexion (P=0.005), plantarflexion (P<0.05) and palpation (P<0.05). These results suggest that some, but not all, CECS patients may be more susceptible to the pain associated with eccentric contractions that is thought to be the cause of damage and inflammation of connective tissue. PMID- 12560957 TI - Assessment of peak power and short-term work capacity. AB - The purpose of this study was to evaluate conditions for conducting a 30 s Wingate test such as load selection, and the method of starting the test (stationary or flying start). Nine male and four female athletes volunteered to be tested on four laboratory visits. Tests were performed on a modified Monark cycle ergometer (Varberg, Sweden) equipped with force transducers on the friction belt and an optical encoder for velocity measurement. Power was calculated with the moment of inertia (I) of the flywheel taken into consideration. One laboratory visit was used to determine individualized optimal resistance conditions. The other three visits were for performance of one of three Wingate tests: a flying start with 0.834 N x kg(-1) [85 g x kg(-1) body weight (BW)] resistance (FLY-0.8); a stationary start with 0.834 N x kg(-1) BW resistance (ST 0.8), or a stationary start with optimal resistance (ST-OPT). FLY-0.8 gave a lower (P<0.05) value for short-term work capacity [19,986 (827) J] than either ST OPT [23,014 (1,167) J] or ST-0.8 [22,321 (1075) J]. Peak power output per pedal revolution was lower ( P<0.005) for FLY-0.8 [833 (40) W] than for either ST-0.8 [974 (57) W] or ST-OPT [989 (61) W]. The results of this study demonstrate that higher values for peak power and short-term work capacity are obtained with a test from a stationary start. It is apparently not necessary to use an individualized optimal resistance when I is considered in a Wingate test initiated from a standstill. PMID- 12560958 TI - Training improves muscle oxidative capacity and oxygenation recovery kinetics in patients with chronic obstructive pulmonary disease. AB - We studied 21 patients with chronic obstructive pulmonary disease aged [mean (SD)] 63 (10) years, with a mean forced expiratory volume in 1 s of 40 (6)% and a peak oxygen uptake of 67 (11)% of predicted values. Patients trained for 6 weeks on a cycle ergometer at high work-rates (WR). Near-infrared spectroscopy was used to obtain the time-constant of the deoxygenation recovery signal (tauHbO(2)) during three constant WR exercise tests, one below and two above the lactic acidosis threshold (theta(L)). Glycolytic and oxidative enzymes and lactate concentrations were assessed in muscle biopsies. The tauHbO(2) decreased significantly in all three constant WR tests: -18 (24)s, -20 (23) s and -13 (22) s, respectively. Endurance time increased in the higher WR tests, by 5.7 (4.8) min and 3.6 (2.7) min, respectively. The activity of citrate-synthase (CS) and creatine-kinase changed significantly from 20 (10) to 30 (13) micro mol x min(-1) x g(-1)and from 3.825 (950) to 3.402 (526) micro mol x min(-1) x g(-1), respectively. Training also improved significantly the mean response time of the on-transient of oxygen uptake (tau'VO(2)) of the below-theta(L) test. We found significant correlations between changes in CS and changes in tauHbO(2), tau'VO(2) and endurance time. We conclude that leg training accelerates the speed of re-oxygenation of the vastus lateralis muscle after exercise. This improvement is correlated to changes in the oxidative enzymes. PMID- 12560959 TI - Energy consumption in static muscle contraction. AB - Energy consumption during static contraction of the human triceps surae muscles was studied in 11 healthy subjects. The subjects had to stand intermittently on the left and then right foot at different frequencies (for periods of 15 s, 10 s or 5 s), first on the whole foot and then on the forefoot. The mean static energy consumption of all subjects was 13.4 (15) W [mean (s.d.)] at a calf muscle moment of 105 Nm. Assuming that static energy consumption (in W) is proportional to static moment (in Nm), a proportionality factor of 0.17 (0.19) s(-1) is found. Because of the limited attainable accuracy, no significant differences between endurance athletes and sprinters could be found. PMID- 12560960 TI - Effect of prior heavy arm and leg exercise on VO2 kinetics during heavy leg exercise. AB - The aim of the present study was to examine the effect of prior exercise at a remote site on the VO(2) kinetics during subsequent heavy cycle exercise using a model that allowed us to discriminate between the VO(2) fast and slow component responses. Ten male subjects completed a constant-load exercise of 6 min cycling at 90% of the VO(2)peak in three conditions: without prior exercise (LE-C), after heavy cycling exercise (6 min at 90% of the VO(2)peak) (LE-L) and after heavy arm cranking exercise (6 min at 90% of the arm VO(2)peak) (LE-A). Subjects performed four repetitions of each exercise protocol, separated by at least 1 day. VO(2) was measured on a breath-by-breath basis and VO(2) kinetics were determined with a biexponential model. There were no significant differences in the VO(2) fast component parameters between LE-C, LE-L and LE-A. However, the VO(2) slow component amplitude was significantly reduced in LE-L and LE-A compared to LE-C, but the reduction was less pronounced in LE-A [the value of the VO(2) slow exponential term at the end of exercise, A(2)', was 657 (SD 200) ml x min(-1) in LE-C versus 384 (SD 136) ml x min(-1) in LE-L and 551 (SD 169) m x min(-1) in LE A; P<0.05]. The results of this study demonstrate that prior heavy arm exercise alters VO(2) kinetics during cycling exercise by reducing the VO(2)slow component amplitude, though this reduction is smaller than the reduction observed following prior heavy leg exercise. These data indicate that the primary factor causing changes in the VO(2) kinetics is probably located in the involved muscle. PMID- 12560961 TI - Transients of the force and surface mechanomyogram during cat gastrocnemius tetanic stimulation. AB - The aim of the study was to investigate the time relationship between force and muscle surface displacement, detected as the surface mechanomyogram (MMG) by a laser distance sensor, in the transient phases of a tetanic stimulation. For this purpose the motor nerve of the exposed medial gastrocnemius of four cats was supramaximally stimulated at 30, 40 and 50 Hz for 9 s. Force was detected by a transducer connected at the distal tendon while MMG was measured after pointing the laser beam at the muscle belly. We found that the MMG always anticipated and trailed the force changes during the on- and off-phase of the tetani, respectively. Independently of the stimulation rate, the half-times of the two signals were: on-phase, about 76 ms for force and 33 ms for MMG; off-phase, about 83 ms for force and 132 ms for MMG. There are two main comments to make about these results. First, during the on-phase the shortening of the contractile elements results at first in a muscle geometry change with low output force. After this, when the slack of the elastic-connective tissue has been taken up, the tension is efficiently transmitted to the tendon. Second, the different force and MMG dynamics in the on- and off-phases determine a counter-clockwise hysteresis with more force produced at a given muscle surface displacement during relaxation. To explain the results, the possible specific roles of some components of the muscle mechanical model, muscle mechanical properties and intra muscular phenomena taking place during contraction have been discussed. PMID- 12560962 TI - Solvent properties of the 1-butyl-3-methylimidazolium hexafluorophosphate ionic liquid. AB - 1-Butyl-3-methylimidazolium hexafluorophosphate (BMIM-PF(6)) was synthesized and purified to be used as a ionic liquid solvent. Its physicochemical properties were studied. The ionic liquid/water (P(il/water)) and ionic liquid/heptane (P(il/heptane)) distribution coefficients of a set of 40 compounds with various functionalities, including organic acids, organic bases, amino acids, antioxidants, and neutral compounds, were measured using liquid chromatography. For ionizable compounds, the P(il/water) values measured at pH 2, 5.1, and 10 were very different. These allowed the determination of both the molecular P(o)il(/water) values and the ion P(-)il(/water) value for each compound. These coefficients were compared to the corresponding P(oct/water) coefficients. Marked differences in the partitioning behavior of basic, acidic, and neutral compounds were observed. The relationship between P(il/water) and P(oct/water) is different from that reported previously. By using the linear free energy solvation approach and the descriptors found for 12 solutes, the BMIM-PF(6) solvent parameters were calculated for the ionic liquid/water and ionic liquid/heptane biphasic systems. The regression parameters show a low basicity of the BMIM-PF(6) solvent compared to octanol. The high cohesion of the ions in the ionic liquid phase is also indicated by the regression equations obtained. Ionized phenols (phenoxide ions) associate more strongly with BMIM-PF(6) than most other ionized molecules. Amino acids were not soluble in ionic liquid; however, it is possible to extract them partially by adding a crown ether to the ionic liquid phase and working at pH 1. The positive form of amino acids is complexed by the crown ether and the complex is extracted in the ionic liquid phase. PMID- 12560963 TI - The pattern of urinary acylcarnitines determined by electrospray mass spectrometry: a new tool in the diagnosis of diabetes mellitus. AB - l-Carnitine and its esters are products of intermediary metabolism of organisms. The distribution pattern or the favored excretion of individual acylcarnitines tells something about metabolic diseases. The determination of the urinary acylcarnitine pattern by flow injection analysis (FIA)-electrospray ionization (ESI)-mass spectrometry (MS) is presented. Groups of healthy probands and patients suffering from diabetes mellitus were investigated due to their significant acylcarnitine profile. The statistical analysis of data sets obtained clearly shows a difference in the acylcarnitine pattern of healthy and sick probands. In comparison to the controls, diabetes mellitus patients excrete more long-chain carnitine esters ranging from dodecanoyl to palmitoylcarnitine. Thus, the urinary acylcarnitine pattern determined by ESI-MS can be a useful tool in the diagnosis and therapy monitoring of diabetes mellitus. PMID- 12560964 TI - Simultaneous determination of domperidone and cinnarizine in a binary mixture using derivative spectrophotometry, partial least squares and principle component regression calibration. AB - This work is concerned with the simultaneous determination of domperidone maleate (DOM) and cinnarizine (CINN) in a binary mixture form, without previous separation, by two different techniques. The first method is the application of derivative spectrophotometry where the linearity range and percentage recoveries for DOM and CINN were 2.5-30 micro g mL(-1), 5-25 micro g mL(-1) and 100.06+/ 1.157, 99.93+/-1.377, respectively. The second method depends on the application of partial least squares (PLS) and principle component regression (PCR) models. A training set consisting of 10 mixtures containing 5-20 micro g mL(-1) for each component was used for the construction of the PCR and PLS models. These models were used after their validation for the prediction of the concentration of DOM and CINN in their mixtures. The proposed procedures were successfully applied for the simultaneous determination of both drugs in laboratory prepared mixtures and in commercial tablet preparations. The validity of the proposed methods was assessed by applying the standard addition technique where the percentage recovery of the added standard was found to be 99.98+/-0.297 and 99.84+/-0.700 for DOM and CINN, respectively, using the derivative spectrophotometric method and 100.29+/-0.398 and 100.11+/-0.363 for DOM and CINN, respectively, using the PLS and PCR methods. The proposed procedures are rapid, simple, require no preliminary separation steps and can be used for routine analysis of both drugs in quality control laboratories. PMID- 12560965 TI - New approach in the treatment of data from an acid-base potentiometric titrationI. Monocomponent systems of monofunctional acids and bases. AB - Based on precise analysis of the acid-base equilibrium, a new approach in the treatment of experimental data from a potentiometric titration is proposed. A new general formula giving explicitly the relation V=f([H(+)]) is derived, valid for every acid-base titration, which includes mono- and polyfunctional protolytes and their mixtures. The present study is the first practical application of this formula for the simplest case, the analysis of one monofunctional protolyte. The collected mV data during the titration are converted into pH-values by means of an auto pH-calibration procedure, thus avoiding preliminary preparation of the measuring system. The mentioned pH-calibration method is applicable also in water organic mixtures and allows the quantitative determination of sparingly soluble substances (particularly pharmaceuticals). The treatment of the data is performed by means of ready-to-use software products, which makes the proposed approach accessible for a wide range of applications. PMID- 12560966 TI - Determination of polychlorinated biphenyl congeners and chlorinated pesticides in a fish tissue standard reference material. AB - The concentrations of a wide range of polychlorinated biphenyl congeners (PCBs) and chlorinated pesticides in a fish tissue Standard Reference Material (SRM) have been determined using multiple methods of analysis. This material, SRM 1946, Lake Superior Fish Tissue, was recently issued by the National Institute of Standards and Technology (NIST) and complements a suite of marine environmental natural-matrix SRMs that are currently available from NIST for the determination of organic contaminants such as aliphatic hydrocarbons, polycyclic aromatic hydrocarbons (PAHs), PCBs, and chlorinated pesticides. SRM 1946 is a fresh tissue homogenate (frozen) prepared from filleted adult lake trout (Salvelinus namaycush namaycush) collected from the Apostle Islands region of Lake Superior. SRM 1946 has certified and reference concentrations for PCB congeners, including the three non- ortho PCB congeners, and chlorinated pesticides. Certified concentrations are available for 30 PCB congeners and 15 chlorinated pesticides. Reference concentrations are available for 12 PCB congeners and 2 chlorinated pesticides. In addition, SRM 1946 is characterized for additional chemical constituents and properties: fatty acids, extractable fat, methylmercury, total mercury, selected trace elements, proximates, and caloric content. The characterization of chlorinated compounds is described in this paper with an emphasis on the approach used for the certification of the concentrations of PCB congeners and chlorinated pesticides. The PCB congener and chlorinated pesticide data are also compared to concentrations in other marine natural-matrix reference materials available from NIST (fish oil, mussel tissue, whale blubber, and a second fresh frozen fish tissue homogenate prepared from filleted adult lake trout collected from Lake Michigan) and from other organizations such as the National Research Council Canada (ground whole carp), the International Atomic Energy Agency (fish homogenate), and the European Commission Joint Research Centre [fish oils (cod and mackerel) and mussel tissue]. PMID- 12560967 TI - Determination of ultratrace elements in natural waters by solid-phase extraction and atomic spectrometry methods. AB - A study was carried out on the preconcentration of ultratrace amounts of cadmium, lead, manganese, copper and iron from high-salinity aqueous samples and determination by atomic spectrometry methods. Sample volume, amount of resin, loading flow rate, and elution volume were optimized in order to obtain the simultaneous preconcentration of all the analytes. Quantitative recoveries were obtained by using 200 mg of iminodiacetic resin with a loading flow rate of 2 mL min(-1), elution volume of 3 mL and sample volume of 50-450 mL. Only copper in seawater samples was not completely retained by the resin (60-70% recovery), due to unfavorable competition of iminodiacetic-active groups with organically bound metal.To quantify the metals in the eluates, two atomic spectrometry techniques were compared: electrothermal atomization atomic absorption spectrometry (ETAAS) and inductively coupled plasma-optical emission spectrometry (ICP-OES) with simultaneous CCD detection system. Both techniques are suitable for sample analysis with detection limits of 1.0, 4.7, 3.3, 6.8, and 53 ng L(-1) using ETAAS and 12, 122, 3.4, 17, and 21 ng L(-1) using ICP-OES for Cd, Pb, Mn, Cu, and Fe, respectively. Relative standard deviations of the procedures ranged from 1.7 to 14% at the sub-microg L(-1) concentration level. The accuracy of both methods was verified by analyzing various certified reference materials (river water, estuarine water, coastal and off-shore seawater). PMID- 12560969 TI - Triton X-100-stabilized "oil-in-water" emulsions as suitable media for alkalimetric determination of hydrophobic organic acids. AB - The protolytic properties of fat row carbonic acids in Triton X-100-stabilized "oil-in-water" (O/W) emulsions were studied. The differential influence of emulsions on the force of acids was shown to depend on the length of the hydrocarbon radical. The conditions for separate alkalimetric determinations of acids with different hydrophobicities in their mixtures using emulsions stabilized by non-ionic surfactant were suggested. The procedure for the determination of 4-chloro- N-(2-furilmethyl)-5-sulfamoylanthranilic acid (furosemide) main substance content in pharmaceuticals using Triton X-100 stabilized emulsion was proposed. PMID- 12560968 TI - Variable selection by an evolution algorithm using modified Cp based on MLR and PLS modeling: QSAR studies of carcinogenicity of aromatic amines. AB - The variable selection in QSAR studies by MLR and PLS modeling has been performed using the evolution algorithm (EA). The Cp statistic has been modified and used as the objective function in the EA search for different combinations of molecular descriptors. For MLR modeling a few information-rich descriptors are selected for model formulation. In PLS modeling, the proposed procedure selects a relatively large number of information-containing descriptors, and a PLS model is formulated based on a few latent variables, which are linear combinations of the selected descriptors. The proposed procedures were used for the prediction of carcinogenicity of aromatic amines. PMID- 12560970 TI - Electrostatic ion chromatography using a carboxybetaine-type zwitterionic surfactant as the stationary phase. AB - A carboxybetaine-type zwitterionic stationary phase obtained by immobilizing Mitsubishi Reagent EF-700 (C(8)F(17)SO(2)NHC(3)H(6)N(+) (CH(3))(2)-C(2)H(4)-COO( )) onto a reversed-phase column was used for chromatographic separation of ions. When aqueous electrolyte solutions having higher pH values (>8) were used as eluents, the model analyte ions (NO(2)(-), H(2)PO(4)(-), Cl(-), Br(-), NO(3)(-), ClO(3)(-), I(-) and SCN(-)) were co-eluted and appeared at the void volume of this HPLC system. However, when aqueous electrolyte solutions having lower pH values (<5.5) were used as eluents, these anions were well retained and separated. Furthermore, when acetate buffers (NaAc/HAc) were used as eluents, plots of log k' (k', retention factor) versus pH of eluents (at constant [NaAc+HAc]), and log k' versus log [NaAc+HAc] (at constant pH), were linear with negative slopes. Breakthrough curves for acid solutions obtained using conductivity detection showed that H(+) ions and their conjugate anions were both retained on the stationary phase and the degree of binding was found to be independent of the acid species used. The degree to which the eluent cation was bound onto the carboxylate functionality of the zwitterion was found to exert a major effect on the retention of analyte anions. A strongly bound cation, such as H(+), reduced electrostatic repulsion effects exerted by the carboxylate functionality on analyte anions, so that they could freely access the quaternary ammonium sites on the zwitterion. It is concluded based on these experimental results that both the charges on the zwitterionic stationary phase make meaningful contributions to the separation of the analyte ions. PMID- 12560971 TI - Preparative separation and determination of matrine from the Chinese medicinal plant Sophora flavescens Ait by molecularly imprinted solid-phase extraction. AB - Molecularly imprinted microspheres (MIMs) were synthesized by micro-suspension polymerization using matrine (MT) as template. The MIMs were employed for solid phase extraction (SPE) and as chromatographic stationary phase for the determination of MT from the Chinese medicinal plant Sophora flavescens. The effects of the various eluents, their concentrations and volumes on the retention behavior were investigated. The selectivity and capacity of the imprinted microspheres against MT was also discussed. The results showed that the MIMs exhibited stronger specific affinity to MT than to oxymatrine (OMT). Methanol water (3:7, v/v) was used for washing impurities from the MIMs-SPE cartridge loaded with the herb extracts, while methanol-glacial acetic acid (9:1, v/v) was used for eluting MT. The maximum load of MT and the recovery of MIMs cartridge towards MT were 38.7 microg g(-1) and 71.4%, respectively. The method developed might be used to separate and extract effective constituents from Chinese medicinal plants on a large scale. PMID- 12560972 TI - On-line complexation/cloud point preconcentration for the sensitive determination of dysprosium in urine by flow injection inductively coupled plasma-optical emission spectrometry. AB - An on-line dysprosium preconcentration and determination system based on the hyphenation of cloud point extraction (CPE) to flow injection analysis (FIA) associated with ICP-OES was studied. For the preconcentration of dysprosium, a Dy(III)-2-(5-bromo-2-pyridylazo)-5-diethylaminophenol complex was formed on-line at pH 9.22 in the presence of nonionic micelles of PONPE-7.5. The micellar system containing the complex was thermostated at 30 degrees C in order to promote phase separation, and the surfactant-rich phase was retained in a microcolumn packed with cotton at pH 9.2. The surfactant-rich phase was eluted with 4 mol L(-1) nitric acid at a flow rate of 1.5 mL min(-1), directly in the nebulizer of the plasma. An enhancement factor of 50 was obtained for the preconcentration of 50 mL of sample solution. The detection limit value for the preconcentration of 50 mL of aqueous solution of Dy was 0.03 microg L(-1). The precision for 10 replicate determinations at the 2.0 microg L(-1)Dy level was 2.2% relative standard deviation (RSD), calculated from the peak heights obtained. The calibration graph using the preconcentration system for dysprosium was linear with a correlation coefficient of 0.9994 at levels near the detection limits up to at least 100 microg L(-1). The method was successfully applied to the determination of dysprosium in urine. PMID- 12560973 TI - A novel analytical low-cost flow system based on a 0.6 MPa (84 psi) diaphragm pump applied to on-line trace pre-concentration in flame AAS and ICP-OES. AB - A novel inexpensive 0.6 MPa (84 psi) flow system based on a low-cost diaphragm pump has been developed. The unfavourable strong pulsation of the pump has been overcome by using highly flexible silicone tubing as a pulse suppression coil. This results in a smooth pulse-free continuous flow of 100 mL min(-1) in circulation. This flow rate is much too high for a flow-injection system; however, with a restrictor capillary the flow rate required can be tapped off down to a range of 0.1-50 mL min(-1). By employing diaphragm pumps in an analytical flow system the pressure gap between HPLC pumps (2-40 MPa) and peristaltic pumps (<0.2 MPa), mainly used in FIA systems, can be filled. Due to the higher pressure delivered by diaphragm pumps relative to peristaltic pumps, the new flow system can be applied to on-line sample pre-concentration and matrix separation in flame AAS and ICP-OES by using standard HPLC pre-columns or small ion-exchange columns, respectively. In this way, very low detection limits in flame AAS have been reached (Cd 0.07 micro g L(-1), Cu 0.05 micro g L(-1), Co 0.9 micro g L(-1), Ni 0.8 micro g L(-1), Mn 0.7 micro g L(-1), Pb 0.8 micro g L(-1) and Tl 0.2 micro g L(-1)). PMID- 12560974 TI - Flow injection determination of papaverine based on its sensitizing effect on the chemiluminescence reaction of permanganate-sulfite. AB - A novel chemiluminescence (CL) method for the determination of papaverine (PAP) has been developed by combining the flow injection technique and its sensitizing effect on the weak CL reaction between sulfite and acidic permanganate. A mechanism for the CL reaction has been proposed on the basis of fluorescent and chemiluminescence spectra. The CL response is proportional to the concentration of PAP over the range 0.2-10 micro mol L(-1). The detection limit of PAP is 0.10 micro mol L(-1) (3 s) with a relative standard deviation (RSD) of 2.0% for 10 repetitive determinations of 1.0 micro mol L(-1) PAP. Interferences from other alkaloids in the opium, such as morphine and codeine, are negligible except that of narcotine. The method has been satisfactorily used for the determination of PAP in injections and compound liquorice tablets. PMID- 12560976 TI - Electrocatalytic oxidation of nitric oxide at indium hexacyanoferrate film modified electrodes. AB - The electrocatalytic oxidation of nitric oxide (NO) by indium (III) hexacyanoferrate (III) films has been studied. These films are electrodeposited onto glassy carbon electrodes through potential cycling in acidic solutions containing a potassium electrolyte, indium (III), and potassium hexacyanoferrate. The resulting modified electrodes exhibit a reversible redox response ascribed to the oxidation/reduction of iron atoms presents in the electrodeposited film. The films have a potent and persistent electrocatalytic activity towards NO oxidation at neutral pH. The electrocatalytic oxidation of NO takes place at potentials around +0.75 V which represents a moderate diminution in the overpotential. In addition, interferences due to the presence of nitrate and nitrite have been significantly reduced. According to these results, the described modified electrodes have been used as sensors for the determination of NO generated by decomposition of a typical NO-donor, such as S-nitroso- N -acetyl- d, l penicillamine (SNAP). PMID- 12560975 TI - Carbon nanotube-enhanced electrochemical DNA biosensor for DNA hybridization detection. AB - A novel and sensitive electrochemical DNA biosensor based on multi-walled carbon nanotubes functionalized with a carboxylic acid group (MWNTs-COOH) for covalent DNA immobilization and enhanced hybridization detection is described. The MWNTs COOH-modified glassy carbon electrode (GCE) was fabricated and oligonucleotides with the 5'-amino group were covalently bonded to the carboxyl group of carbon nanotubes. The hybridization reaction on the electrode was monitored by differential pulse voltammetry (DPV) analysis using an electroactive intercalator daunomycin as an indicator. Compared with previous DNA sensors with oligonucleotides directly incorporated on carbon electrodes, this carbon nanotube based assay with its large surface area and good charge-transport characteristics dramatically increased DNA attachment quantity and complementary DNA detection sensitivity. This is the first application of carbon nanotubes to the fabrication of an electrochemical DNA biosensor with a favorable performance for the rapid detection of specific hybridization. PMID- 12560977 TI - Copper-mercury film electrode for cathodic stripping voltammetric determination of Se(IV). AB - The copper-mercury film electrode has been suggested for the determination of Se(IV) in a wide range of concentration from 1x10(-9) to 1x10(-6) mol L(-1)by square-wave cathodic stripping voltammetry. Insufficient reproducibility and sensitivity of the mercury film electrode have been overcome by using copper(II) ions during the plating procedure. Copper(II) has been found to be reduced and form a reproducible copper-mercury film on a glassy carbon electrode surface. The plating potential and time, the concentration of copper(II) and the concentration of the supporting electrolyte have been optimised. Microscopy has been used for a study of the morphology of the copper-mercury film. It has been found that it is the same as for the mercury one. The preconcentration step consists in electrodeposition of copper selenide on the copper-mercury film. The relative standard deviation is 4.3% for 1x10(-6) mol L(-1) of Se(IV). The limit of detection is 8x10(-10) mol L(-1) for 5 min of accumulation. PMID- 12560978 TI - Determination of copper in coal fly ash in the presence of excess titanium by dynamic reaction cell inductively coupled plasma mass spectrometry. AB - Inductively coupled plasma quadrupole mass spectrometry (ICP-QMS) was used for the accurate determination of copper in coal fly ash samples in the presence of excess titanium, using the reaction of Cu(+) ions with NH(3) in the cell. The method eliminated the effect of polyatomic isobaric interferences at m/z 63 and 65 caused by the formation of (47)Ti(16)O(+), (49)Ti(16)O(+) and (47)Ti(18)O(+) on (63)Cu(+) and (65)Cu(+) by detecting Cu(+) as the product cluster ion Cu(NH(3))(2)(+). As the signal of (63)Cu(NH(3))(2)(+) overlapped with that of (97)Mo(+) which existed in the samples, (65)Cu(NH(3))(2)(+) was detected at m/z 99. The effect of the operating conditions of DRC system was studied in order to obtain the best signal to noise ratio for Cu(NH(3))(2)(+) at m/z 99. The formation of Cu(NH(3))(2)(+) was through the clustering reaction Cu(+)+2NH(3)- >Cu(NH(3))(2)(+) which resulted in the separation of analyte from the interfering oxide. The detection limit for Cu(NH(3))(2)(+) was 0.015 ng mL(-1) as Cu. The method was applied to the determination of copper in NIST SRM 1633a and 1633b coal fly ash reference materials. The precision between sample replicates was better than 2.0% and the analysis results were in good agreement with the certified values. PMID- 12560979 TI - A core-shell structured BaTiO3 precursor preparation, characterization and potential. AB - A new procedure for the preparation of a core-shell-structured BaTiO(3) precursor (core=TiO(2); shell=BaCO(3)) will be described. The structure of this precursor is characterized by electron microscopy (environmental scanning electron microscopy; energy disperse X-ray spectroscopy), whereas the development of phases during thermal treatment is followed by X-ray powder diffraction. PMID- 12560980 TI - Copper(II)-catalysed oxidative coupling reaction of 3-hydroxyacetanilide with 3 methyl-2-benzothiazolinone hydrazone for the spectrophotometric determination of traces of copper(II). AB - A spectrophotometric method is developed for the determination of traces of copper(II), based on the catalytic oxidative coupling reaction of 3 hydroxyacetanilide with 3-methyl-2-benzothiazolinone hydrazone in the presence of ammonia and hydrochloric acid. Beer's law is obeyed in the copper(II) concentration range of 0.008-0.16 microg mL(-1), and the molar absorptivity at 530 nm is 2.5x10(5) L mol(-1) cm(-1). The Sandell's sensitivity of the product is 0.000254 microg cm(-2). The optimum reaction conditions and other important analytical parameters have been investigated. The proposed method is applied to the analysis of water and soil samples and the results are compared with the literature method. PMID- 12560981 TI - A highly sensitive spectrophotometric determination of platinum(IV) using leuco xylene cyanol FF. AB - A new, simple, highly sensitive and rapid spectrophotometric method has been described for the determination of platinum(IV). The method is based on the oxidation of leuco xylene cyanol FF (LXCFF) to its blue form of xylene cyanol FF by platinum(IV) in sulfuric acid medium (pH 1.0-2.5), the formed dye shows an absorption maximum at 620 nm in acetate buffer medium (pH 3.0-4.5). The method obeys Beer's law over a concentration range of 0.3 to 2.6 micro g mL(-1) platinum, having molar absorptivity and Sandell's sensitivity of 5.1x10(4) L mol( )(1) cm(-1) and 0.0038 micro g cm(-2), respectively. The optimum reaction conditions and other analytical parameters have been evaluated. The developed method has been successfully applied to the determination of platinum in pharmaceutical preparations, soil, natural water, plant material, platinum containing catalyst, and synthetic alloy samples. PMID- 12560982 TI - Application of reflectance micro-infrared spectroscopy in coal structure studies. AB - Fourier transform infrared micro-scale analysis of organic matter has been applied to characterise individual petrographic components of coal. The use of an infrared microscope made it possible to study and compare different parts of a single individual grain of vitrinite. This method enables analysis of particular macerals without the need for previous separation from coals. For this purpose the polished grainy micro-sections of coal have been prepared. The results obtained indicated the progression of aromatisation in the vitrinite structure with an increase of coalification degree of the parent coals. PMID- 12560983 TI - Application of chemiluminescence for the detection of peroxy compounds in high performance liquid chromatography. AB - The possibility of applying post-column reaction and chemiluminescence to determine organic peroxy compounds by RP-HPLC was investigated. Conditions of qualitative and quantitative analyses have been established. The applicability of the method has been demonstrated for a series of compounds representative of the most important groups of peroxy-type compounds, that is, hydroperoxides, dialkyl peroxides, diacyl peroxides, peroxyesters, and peroxyacids. PMID- 12560984 TI - Relevance of airborne fungi and their secondary metabolites for environmental, occupational and indoor hygiene. AB - Airborne fungal contaminants are increasingly gaining importance in view of health hazards caused by the spores themselves or by microbial metabolites. In addition to the risk for infection, the allergenic and toxigenic properties, as well as the inflammatory effects are discussed in this review as possible health impacts of bioaerosols. A major problem is the lack of threshold values for pathogenic and non-pathogenic fungi, both in the workplace and in outdoor air. While the relevance of mycotoxins has been intensely studied in connection with contamination of food and feed, the possible respiratory uptake of mycotoxins from the air has so far not been sufficiently taken into account. Toxic secondary metabolites are expected to be present in airborne spores, and may thus occur in airborne dust and bioaerosols. Potential health risks cannot be estimated reliably unless exposure to mycotoxins is determined qualitatively and quantitatively. Microbial volatile organic compounds (MVOC) have been suggested to affect human health, causing lethargy, headache, and irritation of the eyes and mucous membranes of the nose and throat. The production of MVOC by fungi has been discussed in connection with domestic indoor microbial pollution, but the relevance of fungal metabolites in working environments remains insufficiently studied. PMID- 12560985 TI - I do it my way: Regulation of ammonium uptake and ammonium assimilation in Corynebacterium glutamicum. AB - In order to utilize different nitrogen sources and to survive situations of nitrogen limitation, microorganisms have developed several mechanisms to adapt their metabolism to changes in the nitrogen supply. In this communication, recent advances in our knowledge of ammonium uptake, its assimilation, and connected regulatory systems in Corynebacterium glutamicum are discussed with respect to the situation in the bacterial model organisms Escherichia coli and Bacillus subtilis. The regulatory network of nitrogen control in C. glutamicum differs substantially from that in these bacteria, for example, by the presence of AmtR, the unique "master regulator" of nitrogen control, the absence of a NtrB/NtrC two component signal transduction system, and a different sensing mechanism in C. glutamicum. PMID- 12560986 TI - Development of a genetic system for Magnetospirillum gryphiswaldense. AB - Genetic analysis of bacterial magnetosome biomineralization has been hindered by the lack of an appropriate methodology for cultivation and genetic manipulation of most magnetotactic bacteria. In this report, a genetic system for Magnetospirillum gryphiswaldense is described. The system includes a plating technique that allows the screening of magnetic vs non-magnetic colonies, and a protocol for the transfer of foreign DNA by electroporation and high-frequency conjugation. Various broad-host-range vectors of the IncQ, IncP, and pBBR1 groups were found to be capable of replication in M. gryphiswaldense. Several antibiotic resistance markers that can be expressed in M. gryphiswaldense were identified. Tn 5 transposons delivered on a suicide plasmid showed transpositional insertion into random chromosomal sites. PMID- 12560987 TI - Novel carotenoid glucoside esters from alkaliphilic heliobacteria. AB - Pigments of three species of alkaliphilic heliobacteria of the genus Heliorestis, H. daurensis, H. baculata and an undescribed species Heliorestis strain HH, were identified using spectroscopic methods. In these species, bacteriochlorophyll g esterified with farnesol was present, as for other heliobacteria. The carotenoids consisted of 4,4'-diaponeurosporene, also found in other heliobacteria, plus the novel pigments OH-diaponeurosporene glucoside esters (C16:0 and C16:1). In addition, trace amounts of biosynthetic intermediates, OH-diaponeurosporene and OH-diaponeurosporene glucoside, were found. Trace amounts of a carotenoid with 20 carbon atoms, 8,8'-diapo-zeta-carotene, were also found in these species as well as in the non-alkaliphilic heliobacteria. The non-alkaliphilic species Heliophilum fasciatum also contained trace amounts of the two OH diaponeurosporene glucoside esters. The results are used to predict the pathway of carotenoid biosynthesis in heliobacteria. PMID- 12560988 TI - A zinc-containing mannitol-2-dehydrogenase from Leuconostoc pseudomesenteroides ATCC 12291: purification of the enzyme and cloning of the gene. AB - Mannitol-2-dehydrogenase (EC 1.1.1.67) of Leuconostoc pseudomesenteroides ATCC 12291 catalyzing the NADH-dependent reduction of d-fructose to d-mannitol was purified to homogeneity. Native mannitol-2-dehydrogenase has a molecular mass of 155 kDa as determined by gel filtration chromatography. In SDS-PAGE, a single band appeared corresponding to a molecular mass of 43 kDa which indicated that the enzyme was composed of four identical subunits. Enzyme activity was completely inhibited by EDTA and could be restored by zinc ions, but not by Mn(2+) or Mg(2+) which demonstrated that zinc is a cofactor. Purified mannitol-2 dehydrogenase exhibited a maximal specific activity of 400 micromol fructose reduced min(-1) x (mg protein)(-1), using NADH as electron donor. The enzyme showed a high substrate specificity for d-fructose and d-mannitol, however it accepted NADPH as a cofactor with 32% activity ( V(max)) relative to NADPH (100%). The mdh gene, encoding mannitol-2-dehydrogenase, was identified by hybridization with a degenerate gene probe complementary to the nucleotide sequence encoding the first eight N-terminal amino acids of the enzyme. The mdh gene was cloned on a 4.2-kb DNA fragment, subcloned, and expressed in Escherichia coli. Sequencing of the gene revealed an open reading frame of 1017 bp, encoding a protein of 338 amino acids with a predicted molecular mass of 36.0 kDa. Plasmid encoded mdh was functionally expressed, with 70 U/mg of cell-free protein in E. coli. Multiple sequence alignments showed that mannitol-2-dehydrogenase was affiliated with members of the Zn(2+)-containing medium-chain alcohol/polyol dehydrogenase/reductase protein family (MDR). PMID- 12560989 TI - A comparative study of bchG from green photosynthetic bacteria. AB - The gene bchG, coding for bacteriochlorophyll a synthase from a variety of green sulfur bacteria and the filamentous anoxygenic phototrophic bacteria, Chloroflexus aurantiacus, Chloronema sp., and Roseiflexus castenholzii HL08, was partially sequenced and compared. The deduced amino acid consensus sequences for green sulfur bacteria and green filamentous anoxygenic phototrophic bacteria were found to belong to the UbiA enzyme family of polyprenyltransferases with the most similar sequences being those of photosynthetic organisms. All deduced amino acid sequences showed a highly conserved region, which includes the motif DRXXD, characteristic of polyprenyltransferases, which was extended to DREVDAINEP for green sulfur bacteria. Neighbor-joining analysis of a protein similitude matrix displayed a relatively high distance between green sulfur bacteria and the other groups. Sequences from green sulfur bacteria were more closely related to those of purple bacteria than to those of filamentous anoxygenic phototrophic bacteria. In addition, internal grouping within green sulfur bacteria was congruent regarding taxonomic features including cell shape, presence of gas vacuoles and NaCl requirement. In addition to bchlG, another gene encoding for a second chlorophyll synthetase, previously tentatively identified as chlG, was also found in Chlorobium tepidum, showing the highest similarities with polyprenyltransferases from chlorophyll- a-containing organisms. PMID- 12560990 TI - Molecular and biochemical characterization of two tungsten- and selenium containing formate dehydrogenases from Eubacterium acidaminophilum that are associated with components of an iron-only hydrogenase. AB - Two gene clusters encoding similar formate dehydrogenases (FDH) were identified in Eubacterium acidaminophilum. Each cluster is composed of one gene coding for a catalytic subunit ( fdhA-I, fdhA-II) and one for an electron-transferring subunit ( fdhB-I, fdhB-II). Both fdhA genes contain a TGA codon for selenocysteine incorporation and the encoded proteins harbor five putative iron-sulfur clusters in their N-terminal region. Both FdhB subunits resemble the N-terminal region of FdhA on the amino acid level and contain five putative iron-sulfur clusters. Four genes thought to encode the subunits of an iron-only hydrogenase are located upstream of the FDH gene cluster I. By sequence comparison, HymA and HymB are predicted to contain one and four iron-sulfur clusters, respectively, the latter protein also binding sites for FMN and NAD(P). Thus, HymA and HymB seem to represent electron-transferring subunits, and HymC the putative catalytic subunit containing motifs for four iron-sulfur clusters and one H-cluster specific for Fe only hydrogenases. HymD has six predicted transmembrane helices and might be an integral membrane protein. Viologen-dependent FDH activity was purified from serine-grown cells of E. acidaminophilum and the purified protein complex contained four subunits, FdhA and FdhB, encoded by FDH gene cluster II, and HymA and HymB, identified after determination of their N-terminal sequences. Thus, this complex might represent the most simple type of a formate hydrogen lyase. The purified formate dehydrogenase fraction contained iron, tungsten, a pterin cofactor, and zinc, but no molybdenum. FDH-II had a two-fold higher K(m) for formate (0.37 mM) than FDH-I and also catalyzed CO(2) reduction to formate. Reverse transcription (RT)-PCR pointed to increased expression of FDH-II in serine-grown cells, supporting the isolation of this FDH isoform. The fdhA-I gene was expressed as inactive protein in Escherichia coli. The in-frame UGA codon for selenocysteine incorporation was read in the heterologous system only as stop codon, although its potential SECIS element exhibited a quite high similarity to that of E. coli FDH. PMID- 12560991 TI - Expression of the iron-responsive irpA gene from the cyanobacterium Synechococcus sp strain PCC 7942. AB - Expression of the iron-stress-induced irpA gene of Synechococcus sp. strain PCC 7942 was investigated by constructing luminescent p irpA::luxAB promoter fusions. Growth of Fe-replete and Fe-limited cultures yielded high levels of luminescence only under conditions of iron deficiency. Promoter fusion deletions revealed that low Fe irpA transcription is dependent on a 25-nucleotide sequence that includes a region of dyad symmetry centered 19 nucleotides from the transcription start. Assaying luminescence at defined iron concentrations in trace-metal-buffered media showed that irpA transcription is activated at concentrations below 100 nm Fe. Overall, the expression pattern and promoter structure of irpAsuggests a novel form of metal-dependent regulation in this species. PMID- 12560992 TI - Growth of sulfate-reducing bacteria and methanogenic archaea with methylated sulfur compounds: a commentary on the thermodynamic aspects. AB - Methylated sulfur compounds such as dimethylsulfoniopropionate, dimethylsulfide, methanethiol, and other methylated sulfur compounds can act as sources of carbon and energy for the growth under anoxic conditions of a number of sulfate-reducing bacteria and methanogenic archaea. We summarise the range of degradative reactions that do or might occur in such organisms, and present thermodynamic data for these processes. These data enable estimates of the feasibility of the reactions as growth-supporting systems, and of the possible maximum growth yields of the bacteria and archaea catalysing them. We compare our new estimates with the few data that are currently available from the literature, and show that some published growth-yield assessments need reevaluation. PMID- 12560994 TI - [The Family Conference as an example of involving the patient's relatives in neurological early rehabilitation]. AB - For the patient the social support plays a crucial role when trying to cope with critical life events. Therefore the "Saarland-Heilstatten" hospitals in Saarbrucken usually integrate the relatives of patients in a minimally conscious state ("apallisches Syndrom") or having a severe brain injury into the therapies during the neurological early rehabilitation. All members of the interdisciplinary working team attend to the relatives of those patients: there are one-to-one meetings with therapists, weekly meetings with groups of relatives, and Family Conferences. This article explains why generally the qualified work with patients' relatives is absolutely necessary, and it illustrates that concept by describing the approach of the Family Conference. Finally the therapeutic possibilities and limitations of Family Conferences in the rehabilitation process are discussed as seen after several years of experience. PMID- 12560995 TI - [Aptitude statements after head injury]. AB - In an exemplary way the case study of M. S. shows that the statements concerning aptitude, which are based on intelligence structure tests, lead to false positive judgements under certain preconditions. Particularly if you postulate learning disorder and partial amnesia, the standard status diagnosis via intelligence and achievement tests has to be supplemented by a method of testing by which the dynamics of cognitive processes can be recorded. In the individual case study of M. S., learning disorder and partial amnesia have been disclosed via the dynamic learning test by Seitz (1998). They did not become manifest when only the routine diagnostic methods were applied. Against the background of socio-legal implications of the specific case some demands on psychological diagnosis and valuation are being discussed from the practician's point of view. They subsequently lead to certain demands concerning academic research. This applies above all to the dynamic learning test which is described above. It states as a result a linear relationship between the effort put into learning and the quantity of learning (the slope is interpreted as an individual constant). The dynamic learning test is also likely to be interesting in respect to pure research in general psychology and the formation of theories. PMID- 12560996 TI - [The occupational therapy assessment: implementation in practice]. AB - The "Ergotherapeutische Assessment" (EA, occupational therapy assessment instrument) is designed as a generic instrument assessing disabilities with consequences on everyday life. It is tested for its practicability and measurement properties and suitable for adult patients treated with occupational therapy. It can be used for routine documentation, as a result-indicator for data based quality management and in outcome research. The EA is integrated in an assessment network currently in development intended for occupational therapy, physiotherapy, speech therapy and, optionally, for further professions. The profession-specific approach should enhance the intrinsic motivation for self evaluation and thus the learning organisation. PMID- 12560997 TI - [Individualized outcome measurement -- a comparison between prospective and retrospective problem marking in psychosomatic rehabilitation]. AB - This article reports on a questionnaire concerning individual problems in psychosomatic rehabilitation. A sample of 858 patients answered the questions at the beginning and on a one year catamnestic measurement. First, the frequency of occurrence of several symptoms was asked. Second, the rehabilitants were asked to indicate for each symptom if it was an important problem before treatment. This procedure of identifying problems was done at the beginning (prospective marking) and also at the one year follow up (retrospective marking). The questionnaire allows to compute eight subscales and one global scale regarding all items (classical approach). Frequency of occurrence decreased with an average effect size (ES) of 0.56 on the global scale. In the next step, measurement was individualized by regarding only items which were marked as relevant problem. By prospective marking, effect size was 1.32. By retrospective marking, no growth of effect size in comparison to the classical approach was found (ES = 0.56). The results indicate that the prospective approach of identifying problems is the better way of individualizing measurements. Moreover, the impact of individual problem definition on outcome was demonstrated. PMID- 12560998 TI - [Occupationally Orientated Medical Rehabilitation (BOR) for disabilities caused by orthopedic diseases]. AB - Low back pain and arthrosis are influencing the quality of life of people concerned and are leading to high expenses for the health and social system. A merely medical rehabilitation is often insufficient in case the person concerned is unable to fulfil the requirement at work. But vocational rehabilitation is often connected with long processing and waiting times. To counteract these, cooperative approaches have been developed between rehabilitation clinics and vocational retraining centres (Berufsforderungswerke, BFW) over the last few years. The Bad Eilsen rehabilitation centre of the LVA Hannover has worked with the Bad Pyrmont BFW since 1998. According to this project, "Occupationally Orientated Medical Rehabilitation" (Berufsorientierte medizinische Rehabilitation, BOR), the participants are extensively informed and tested in the retraining centre, they can also perform occupation-specific tasks. The aim of the study was to compare processing and waiting times with and without BOR. The results available so far show that a majority of the participants of the BOR measures could be occupationally reintegrated. In many cases work promotion measures were introduced earlier. BOR measures not only lead to a considerable acceleration of procedures, they also improve the occupational reintegration of the insured to a high degree. PMID- 12560999 TI - [Vocational rehabilitation in German vocational retraining centres and results of Berufsforderungswerk Heidelberg gGmbH]. AB - The article focuses on the results of vocational integration over the last two decades and on actual results from client-surveys as presented by the Arbeitsgemeinschaft Deutscher Berufsforderungswerke (ADB) (working group of the German vocational retraining centres) and the Berufsforderungswerk Heidelberg (BFW Heidelberg, the vocational retraining centre in Heidelberg). Process quality of vocational rehabilitation can be indicated by data like the number of persons dropping out of their training - the ratio to be found in the vocational retraining centres is on a lower level than in other institutions of vocational training. Clients have changed: during the last decade the average age of the clients in the BFW Heidelberg has increased by 3 years, there is a higher proportion of women, but there also are more clients with psychological disorders. A new group of clients is growing - non-handicapped persons who pay for their training themselves. Process quality has improved, with graduates of the BFW Heidelberg saying significantly more often that they can use much or very much of the training courses' contents within their later occupational activities -- the proportion increases with the level of training they had undergone. Additionally the proportion of those is growing who see themselves in an -- according to their training -- adequate occupational situation. These are certainly positive results referring to the indicators of process quality. Concerning outcome quality one has to take the levels of occupational integration into account. Compared with the results of German vocational retraining centres overall, graduates of the Heidelberg centre constantly had a higher level of vocational integration over the last 20 years, one of the reasons being that the vocational training offered in Heidelberg reaches up to the level of degree of a university of applied sciences. The situation on the job market strongly influences occupational integration: it is evident that a high rate of unemployment will extend the time span necessary for placement. But in the long run occupational integration proves to be amazingly high. Variables that influence the results are: level of disability, sex, age and marital status. The level of vocational qualification is a characteristic with high evidence: integration of graduates at the level of "Fachschule" (highly skilled technician or the like) is 10 % higher and that of graduates at the level of "Fachhochschule" (university of applied sciences) another 10 %. Participation in vocational rehabilitation generally is highly estimated -- estimation as given two years after the training increases with the level of qualification. In the Heidelberg centre progress is seen in expanding the proportions of special target groups (which of course is politically aimed at): women now account for some 30 %, there are more elderly clients and for example also more clients with psychological disorders. The vocational integration of these various groups has to be regarded under different aspects. For example, the situation of severely disabled clients is quite ambivalent: their proportion is falling and their rate of occupational integration is relatively low -- yet there are effects that may compensate. So the importance of the disability decreases with a rising level of vocational training. The data presented may permit the conclusion that vocational rehabilitation at the Heidelberg centre has become more successful over the last decade. PMID- 12561000 TI - [Outpatient geriatric rehabilitation: an evaluation of two models assessing trends of medical outcomes]. AB - In Germany, complex and intensified outpatient geriatric rehabilitation is currently scarcely practised, mainly in model projects. The evaluation of these projects is exclusively conducted in uncontrolled studies. In our project "AMBRA", two different organisational models of geriatric rehabilitation are compared: a mobile rehabilitation team based at a geriatric hospital department and an outpatient rehabilitation centre run by GPs trained in geriatrics. Outcomes were assessed in terms of capability of self-care (Barthel-Index), mobility (Tinetti-Test, Timed "Up & Go"-Test, TUG), and depression (Geriatric Depression Scale, GDS). They were documented at three points in time (start of rehabilitation, end of rehabilitation, 6 months after end of rehabilitation) and analysed by multivariate analyses of variance (repeated measurements). 162 complete patients histories were taken in the first 18 months of the project. They show significant improvements in capability of self-care and mobility (both Tinetti-Test and TUG) between the beginning and the end of rehabilitation (adjusted for age, sex, cognitive function, diagnosis, rehabilitation model). On a medium-term basis, these results remained stable (TUG declined, however). Average GDS values did not change significantly. There were no significant sex- or age-related effects. The patients' cognitive function influenced changes in the results of the Barthel-Index and the Tinetti-Test. Patients with skeletal diseases showed less favourable trends in the Barthel-Index as did patients with cognitive impairments caused by vascular disease in the TUG, but these patients also benefited in the course of the model rehabilitation procedures. Differences in trends between patient groups of the two models were observed in the Barthel Index. 96 % of patients previously living at home were still living there at the end of rehabilitation, 91 % were still living there 6 months after the end of rehabilitation. At the end of rehabilitation, 67 % of patients described an improvement of their personal situation associated with the rehabilitation procedure. Six months later, 82 % described an improvement or a stabilisation of their personal situation. Our results show positive medium-term rehabilitation trends concerning medical and subjective outcomes. In order to analyse effectiveness, we will have to wait for the results of a regional control group which is being recruited. PMID- 12561001 TI - [The practice of needs assessment for the supply with technical aids in The Netherlands]. AB - In Germany the assessment of needs for technical aids occurs widely unsystematic and without the possibility to verify the advice process. A systematic assessment of individual needs for technical aids that follows standardised indication criteria so far has not been paid much attention. Compared with this, the advice and supply measures in the Netherlands are increasingly based on a systematic and comprehensible analysis of the individual inquiry. This analysis is called "indication and selection advice" and is usually carried out by therapists. Under Dutch law, consultants can act rather independently of any commercial interest. The basics of the analysis derive from the ICIDH and theoretical models of occupational therapy. Besides an overview of the legislation on the supply with technical aids in the Netherlands, the procedure of needs assessment plus a specific instrument based on occupational therapy knowledge are presented. The realization of advice as a defined part of the supply process required a new definition of tasks for occupational therapists. The resulting changes in the comprehension of professional roles are briefly described, too. The results of this investigation regarding the practice of needs assessment in the Netherlands suggest that changes are also required in the German service delivery system and its legal basis. Because of their specific expert knowledge of indication and selection of suitable technical aids, a more extensive involvement of occupational therapists in this key process of measures of home adaptation and the supply with technical aids is recommended in Germany, too. The development of practicable, standardised instruments to assess individual needs is advisable because it will contribute to increased efficiency of the supply with technical aids. PMID- 12561003 TI - Preparation, premedication and surveillance. AB - The main end points for sedation during endoscopy are patients' satisfaction, short duration of the procedure, and safety. During the last year, attention has focused on attempting to identify the "ideal" candidate for moderate sedation/analgesia and on the importance of providing the patient with appropriate information before the procedure. The increasing pressure to perform more procedures, reduce costs, and achieve shorter patient turnaround times has affected recent approaches to sedation during endoscopy, focusing attention on alternatives to pharmacological sedation such as providing relaxing music, using small-caliber endoscopes for unsedated peroral gastroscopy, and using magnetic endoscopic imaging to increase tolerance and reduce discomfort during colonoscopy. The results, however, have not been convincing. The role of benzodiazepines was discussed in some studies, highlighting the well-known effect of midazolam on postprocedural amnesia, its pharmacological profile and tolerability after intranasal spraying in healthy volunteers, and the efficacy and safety of this route of administration as an alternative to intravenous administration in diagnostic upper gastrointestinal endoscopy. The form of sedation for gastrointestinal endoscopy that has attracted great interest over the last year is the use of intravenous propofol, either alone or with concomitant benzodiazepines or opioids. As expected in view of the drug's known pharmacological properties, the quality of sedation was better and recovery time was shorter in patients treated with propofol. However, important questions involving the narrow therapeutic range and the mode of administration of propofol (by endoscopists or nurses, or by anesthesiologists) remain open. One important aspect of sedation procedures is prevention of cardiopulmonary complications. The use of electronic monitoring techniques, with a pulse oximeter, has been recommended as a standard procedure during digestive endoscopy; however, pulse oximetry no longer reflects the normal ventilatory functions and does not detect episodes of severe CO2 retention. CO2 monitoring by transcutaneous measurement - or better, by capnography - appears to be useful, as an alternative to pulse oximetry, as a measure of hypoventilation, and for detecting potentially important abnormalities in respiratory activity in patients undergoing sedation for gastrointestinal endoscopy. With regard to preparation for endoscopic procedures, several "ideal" formulas for bowel preparation have been presented. These include the use of sodium phosphate compounds as an alternative to polyethylene glycol electrolyte lavage solutions (PEG-ELS); however, the results so far have been conflicting. The best and most cost-effective bowel cleansing procedure for colonoscopy and sigmoidoscopy has yet to be established. PMID- 12561004 TI - Reflux disease and Barrett's esophagus. AB - Gastroesophageal reflux disease (GERD) is one of the most prevalent gastrointestinal disorders. The key feature of GERD is reflux of gastric contents into the esophagus. Medical treatment with proton-pump inhibitors (PPIs) is well established and is considered the standard treatment. Given the high prevalence of the condition and the excellent response to medical therapy, antireflux surgery is an option for patients with volume reflux that is not properly controlled by medical therapy. Adenocarcinoma is a rare but life-threatening complication of GERD. The only known precursor lesion for esophageal adenocarcinoma is Barrett's esophagus. In recent years, a clearer understanding of the development of Barrett's and of its progression toward invasive cancer has developed. Genetic factors almost certainly determine the individual risk. The length of the Barrett's esophagus segment and the size of a hiatal hernia are associated with the risk of developing high-grade dysplasia and esophageal adenocarcinoma.With regard to the clinical management of GERD patients with Barrett's, endoscopic surveillance at 3-year intervals is now considered appropriate in the absence of dysplasia. In patients with high-grade dyspepsia, the situation is more difficult. While a considerable proportion of these patients may already have invasive cancers, there is also the possibility that there is only focal dysplasia. For this reason, it is justifiable to carry out curative endoscopic resection. Mucosal ablation procedures may also be appropriate, but these still need to be properly investigated in clinical trials. PMID- 12561005 TI - Treatment of esophagogastric tumors. AB - Esophageal and gastric tumors are often considered as a single group: they share similar symptoms - upper GI endoscopy with a flexible video-endoscope is the gold standard procedure of detection - similar techniques of endotherapy for cure or palliation are offered for both types of tumors. When the endoscopic procedure is performed for a superficial cancer or its precursors, with a curative intent, endoscopic mucosal resection (EMR) is generally preferred to mucosal ablation with a thermal (Nd:YAG) or non-thermal (photodynamic therapy) procedure. In addition to esophageal squamous cell cancer and gastric cancer, new indications of EMR arise in the Barrett esophagus. Guidelines for safe indications concern diameter, polypoid or non polypoid morphology with the subtypes elevated, flat and depressed, and depth of invasion. A superficial invasion in the sub-mucosa is a relative contra-indication in the esophagus, but not in the stomach. The technique of EMR is now codified with an injection into the submucosa for lifting the lesion and either suction with a cap, grasping with a forceps if a 2 channel instrument is used, or tissue incision with a needle knife. En bloc, gives better results than piecemeal resection. The most frequent complication is bleeding. When legitimate indications are respected, the results of EMR are equivalent to those of surgical resection and have reached the consensus level. The major indication in palliation is the relief of dysphagia from malignant esophageal obstruction. Increased indications are proposed for malignant pyloric obstruction. Multiple models of metal expandable and coated stents with appropriate balance between rigidity and flexibility (nitinol alloy) and enough expansive radial force are now offered. After stenting the survival period is short and there is a toll of complications. PMID- 12561006 TI - Endoscopic ultrasonography. AB - This review again uses a practical arrangement of the topics of endoscopic ultrasonography (EUS) into clinical categories rather than single diseases. The clinical categories include established indications, such as staging of gastrointestinal tract cancers, differential diagnosis of submucosal tumors, evaluation of solid and cystic pancreatic masses, detection of lymph nodes, evaluation of the biliary tree, and fine-needle aspiration (FNA). In addition to established indications, surgical and investigational techniques are presented, such as fine-needle therapeutic procedures and methods of studying portal hypertension and hepatic, abdominal, and retroperitoneal masses. This year, the new techniques and instruments presented have not included new probes or endoscopes, as these have already gained wide application in published papers and clinical practice. However, new techniques capable of facilitating the use of existing equipment are discussed, such as computer-aided EUS image analysis. Finally, some fascinating studies investigated outcomes with EUS and the cost effectiveness of the method - stimulating both endosonographers and non endosonographers to think about current practices and ways of improving them. PMID- 12561007 TI - Variceal bleeding and portal hypertension: has there been any progress in the last 12 months? AB - A review of the literature on the management of esophagogastric varices published in the last 12 months shows that the data are still quite conflicting. In the primary and secondary prophylaxis of variceal bleeding, beta-blockers are still the mainstay of pharmacotherapy. Measurement of the hepatic portal venous pressure gradient is considered to be a reliable parameter for successful reduction of portal pressure using medical therapy. However, intolerance of propranolol requiring discontinuation of therapy has been observed in approximately 30 % of patients. Patients' compliance with medication may represent another drawback of medical therapy. The role of endoscopic band ligation in secondary prophylaxis is now indisputable, especially in comparison with sclerotherapy. In the primary prevention of variceal bleeding, band ligation is beginning to have a competitive edge over pharmacological therapy. Acute variceal bleeding is no longer a frequent morbid emergency. Most cases of bleeding can now be managed successfully with band ligation and N-butyl-2 cyanoacrylate obliteration. N-butyl-2-cyanoacrylate has come into increasingly widespread use in the treatment of bleeding gastric fundal varices in which surgery or transjugular intrahepatic portosystemic shunting were previously regarded as the preferred therapies. PMID- 12561008 TI - Diagnostic cholangiopancreatography. AB - As in recent years, the articles on diagnostic endoscopic retrograde cholangiopancreatography (ERCP) published between July 2001 and June 2002 again reflect a defensive attitude and are mainly concerned with magnetic resonance cholangiopancreatography (MRCP) and the complications associated with the new imaging method. Diagnostic ERCP is declining in importance and has held its position mainly due to new advances in tissue sampling, pressure measurement, and the use of echo probes and miniature endoscopes. Problems of training in ERCP have still not yet been resolved, due to the contradictions inherent in low case loads, the desire to offer a wide spectrum of training, and issues of quality assurance. PMID- 12561009 TI - Therapeutic biliary endoscopy. AB - Some steps of progress have been seen in the area of biliary therapeutic endoscopy, in the understanding and management of primary sclerosing cholangitis, problems relating to liver transplantation, malignant biliary strictures, complications after endoscopic retrograde cholangiopancreatography, sphincter of Oddi dysfunction, and tissue sampling. The benefits (or lack thereof) of preoperative biliary drainage in the setting of malignant obstructive jaundice have received an extensive review. PMID- 12561010 TI - Inflammatory bowel disease. AB - Significant advances have been made in our understanding of the etiology of inflammatory bowel diseases, particularly with the identification of the first gene mutation associated with Crohn's disease; the treatment arsenal has also been expanded, and some new developments have been seen with regard to diagnosis as well. The importance of endoscopic features in relation to the prognosis has been extensively studied. New ultrasound techniques and magnetic resonance-based imaging are on the verge of modifying the diagnostic approach, particularly during follow-up in Crohn's disease of the small bowel. By contrast, virtual colonoscopy using computed tomography or magnetic resonance imaging does not appear to be of much help. However, the development of these new techniques is continuing rapidly, and an explosion of new information may be expected in the coming years. PMID- 12561011 TI - Minimally invasive surgery. PMID- 12561012 TI - Covered expandable metallic stent placement for hemostasis of colonic bleeding caused by invasion of gallbladder carcinoma. AB - A 72-year-old Japanese man was admitted to our hospital complaining of right upper-quadrant abdominal pain, blood in his stool, and symptoms of anemia. On physical examination a hard mass, about 6 cm in diameter, was palpable in the right upper quadrant of the abdomen. Computed tomography revealed a gallbladder carcinoma which had invaded the transverse colon, with liver metastasis. We diagnosed gallbladder carcinoma, stage IVB. Colonoscopy was performed for persistent blood in the stools. This revealed an elevated lesion which appeared to be an invasion of gallbladder carcinoma, with diffuse bleeding from the right side of the transverse colon. It proved difficult to stop this bleeding by ordinary therapeutic endoscopy. In order to achieve hemostasis we therefore inserted a covered Ultraflex metallic stent to compress the tumor. After stent placement, blood was no longer seen in the patient's stools, he became able to eat soft food and was discharged. This treatment was uninvasive and effective. Covered stent placement appears to be a new and useful method in the management of bleeding from malignant gastrointestinal tumors. PMID- 12561013 TI - Newly developed 3D endoscopic system: preliminary experience. AB - Although there have been earlier reports on the use of three-dimensional endoscopic systems in the gastrointestinal tract, real-time three-dimensional imaging has not previously been described. This paper presents experience in three cases with a novel endoscopic system for three-dimensional observation in clinical situations. Case 1 was a hyperplastic gastric polyp, the shape of which was considerably enhanced in comparison with two-dimensional endoscopic viewing. Case 2 was an advanced esophageal cancer, in which the sense of depth was improved in the esophagus. Case 3 was a depressed early gastric cancer, the shape of which was also enhanced. Between July 2001 and March 2002, three experienced endoscopists also used this new system in a further 34 patients. A three dimensional endoscopic environment was visible with the system in all patients, and all of the endoscopists were able to obtain visual information regarding spatial depth. This experience suggests that the three-dimensional video endoscopic system used here can be of value in gastrointestinal endoscopy. PMID- 12561014 TI - A case of gallbladder carcinoma diagnosed preoperatively by the detection of human telomerase reverse transcriptase mRNA in endoscopically obtained gallbladder bile. AB - It is difficult to accurately diagnose the nature of polypoid lesions in the gallbladder. To increase diagnostic accuracy, we have developed an endoscopic technique for obtaining gallbladder bile, termed endoscopic transpapillary catheterization into the gallbladder (ETCG). We describe here a case of gallbladder carcinoma diagnosed preoperatively by the detection of human telomerase reverse transcriptase (hTERT) mRNA, a catalytic subunit of telomerase, in gallbladder bile obtained using the ETCG technique. A patient with a polypoid lesion 15 mm in diameter in the gallbladder was admitted to our hospital for further examinations. Gallbladder bile collected by the ETCG technique was evaluated cytologically and also analyzed for telomerase activity and hTERT mRNA. Although the results for cytology and telomerase activity were negative, that for hTERT mRNA was positive. Open surgery was carried out and it was confirmed that the lesion was an adenocarcinoma invading the subserosa. The molecular biological analysis of gallbladder bile collected using the ETCG technique was shown to be effective for diagnosing the nature of the polypoid lesion in the gallbladder. PMID- 12561015 TI - Use of an endoscopic clipping device to repair a duodenal perforation. PMID- 12561016 TI - A potentially serious disadvantage of multiband device over conventional ligation using an overtube. PMID- 12561017 TI - Esophageal dislodgment of a variceal ligator cap due to size mismatch between the ligator cap and the endoscope. PMID- 12561018 TI - Small bowel obstruction and perforation after colonoscopy. PMID- 12561019 TI - Silastic band migration: an unusual complication following vertical banded gastroplasty. PMID- 12561020 TI - Ascariasis observed by wireless-capsule endoscopy. PMID- 12561021 TI - Correlation of number of intrathyroid lymphocytes with antimicrosomal antibody titer in Hashimoto's thyroiditis. AB - The variants of Hashimoto's thyroiditis (classic, florid, and fibrotic types) differ in clinical manifestations, laboratory tests, and morphology. The titers of autoantibodies correlate with the degree of thyroidal lymphocytic infiltration (Wislon et al., 1998(5)) It appears that the density of lymphocytes is inversely proportional to antimicrosomal antibody (AMA) titer. The aim of the present study is to determine the precise relationship between the number of lymphocytes and AMA titer. It may be useful to predict the onset of the disease in subclinically hypothyroid patients. In 62 cases of cytologically confirmed Hashimoto's thyroiditis, the number of lymphocytes per high-power field was counted and correlated with AMA titer. The grading of lymphocytes was done as 1 (<10/HPF), 2 (11-20/HPF), and 3 (>/=21/HPF). The results showed no correlation between lymphocyte number and AMA titer. PMID- 12561022 TI - Distribution of subtypes of metastatic renal-cell carcinoma: correlating findings of fine-needle aspiration biopsy and surgical pathology. AB - Clear-cell (CRCC), papillary (PRCC), and chromophobe (CHRCC) renal-cell carcinoma (RCC) are the three most frequent subtypes of RCC. The rate and distribution of their metastatic lesions have not been well studied in cytopathological materials. Sixty-two fine-needle aspiration biopsy cases of metastatic RCC were studied and correlated with surgical pathology of RCCs with and without metastasis. Special stains for glycogen and immunostaining for cytokeratins, vimentin epithelial membrane antigen (EMA), and carcinoembryonic antigens, and electron microscopic studies were performed. Fifty-nine cases of CRCC and three of PRCC subtypes were retrieved from the cytopathology files at the Ottawa Hospital in a period of 10 years. Of these cases, 10 metastatic CRCC and one metastatic PRCC were diagnosed prior to the diagnosis of the primary tumor. CHRCC and sarcomatoid RCC were not represented in cytopathological specimens. CRCC displayed characteristic filmy cytoplasm and nuclei with prominent nucleoli. PRCC was characterized by dense cytoplasm, large nuclei with prominent nucleoli, and papillary architectures. In addition, all RCCs were characterized by the presence of glycogen and the absence of mucin by using histochemical techniques and electron microscopic studies and positive reactivity for cytokeratins (CK) and vimentin (VIM). In the same period, there were a total of 380 patients with RCC divided into 310 CRCCs, 55 PRCCs, and 15 CHRCCs associated with metastases in 142, 9, and 1 case, respectively. CRCC is by far the most common subtype found in metastases sampled in cytopathology. PRCC, CHRCC, and sarcomatoid RCC were underrepresented. Awareness of this propensity of RCC and the characteristic cytopathological, histochemical, immunohistochemical, and ultrastructural features are helpful in the diagnosis of metastatic RCC. PMID- 12561023 TI - Ciliated epithelial cells in Warthin's tumor on fine-needle aspiration. PMID- 12561024 TI - Solitary parotid metastasis from columnar cell carcinoma of the thyroid: a diagnostic dilemma. AB - A 65-yr-old woman presented in May 1998 with a left parotid mass of 2 yr duration. The cytology and histopathology of the parotid lesion showed features of a columnar cell carcinoma. In November, 1967, she was diagnosed to have a classic papillary thyroid carcinoma (PTC). Ultrasound examination revealed a persistent thyroid nodule, the aspiration cytology of which confirmed columnar cell variant of papillary thyroid carcinoma (CCV-PTC). CCV-PTC is a recently described rare tumor of the thyroid that has been associated with an aggressive clinical course. Our case was peculiar in that the patient presented with an isolated metastasis in the parotid gland. Second, after an indolent course following the initial diagnosis of usual PTC 31 yr ago, she had now developed a more aggressive variant of the tumor. PMID- 12561025 TI - Fine-needle aspiration cytology of chondroblastoma of soft parts: case report and differential diagnosis with other soft tissue tumors. AB - Chondroblastoma is a benign tumor arising in the epiphysis of long bones. The extraskeletal presentation is most unusual. We report the first cytological description of a soft tissue chondroblastoma. It was a subcutaneous mass in the leg of a 62-yr-old man. Fine-needle aspiration (FNA) rendered a highly cellular material with grouped and single polygonal or round cells with a uniform, sometimes eccentric nucleus. Microvacuolated cytoplasm and hemosiderin pigment were frequent findings. There were rare nuclear grooves and mitoses. A metachromatic, focally calcified stroma was present, occasionally surrounding the cells. There were also numerous multinucleated osteoclast-like giant cells. Histological evaluation was diagnostic of chondroblastoma. The tumor was locally aggressive. A review of other soft tissue masses with similar cytological findings is included in the discussion. FNA cytology is very helpful in the diagnosis of soft tissue chondroblastoma, but additional studies may be necessary for a definitive diagnosis. PMID- 12561026 TI - Fine-needle aspiration cytology of splenic hamartoma. AB - Bench-top fine-needle aspiration biopsy (FNAB) following splenectomy and nephrectomy from a 21-yr-old female of tuberous sclerosis with right renal angiomyolipoma and splenic hamartoma was performed for the correlation of the cytologic features with histologic findings and, along with the immunopathologic studies, for the establishment of a challenging preoperative diagnosis of splenic hamartoma. The postoperative bench-top aspirate of splenic hamartoma yielded small and large clusters of plump, spindly to polygonal cells in the blood stained background with scattered small lymphocytes, comparable with the preoperative echo-guided FNAB. The histopathology and immunopatholgy of the splenic hamartoma from the bench-top aspirate cytoblock and splenectomy tissue were comparable, unveiling the red pulp tissue consisting of wider cord stromal tissue than its normal counterpart with sinus-like vascular channels lined by endothelial cells which were positive for CD8, CD34, factor VIII-related antigen, vimentin, as well as negative for muscle-specific actin, CD21, CD68, and cytokeratin, scattered dispersed CD3- and CD45RO-positive T-lymphocytes, and small clusters of CD20-positive B lymphocytes, with a dearth of fibrous trabeculae and organized white-pulp lymphoid tissue. A combination of cytologic features and immunopathologic results from the aspirate cytoblock along with the salient clinical information should enable an accurate preoperative diagnosis of splenic hamartoma. PMID- 12561027 TI - Cell cannibalism: diagnostic marker of malignancy. AB - In this study, we tried to find out the cytological relevance of cannibalism as a dependable feature of malignancy in effusion and urine cytology. We randomly selected a total of 40 cases consisting of 10 each of malignant effusion, benign effusion, malignant urine samples, and benign urine samples. These smears were assessed for the presence of cell cannibalism. The number of cannibalistic cells/100 tumor cells was counted. The cannibalistic cells were seen more commonly in malignant effusion cases (3.4/100 cells) compared with malignant urine cases (2/100 cells). There was not a single cannibalistic cell in benign conditions. The finding of an increased number of cannibalistic cell was highly significant in malignant versus benign samples (P > 0.0000, Student's t-test). The present study highlights the significance of cannibalism in malignant urine and effusion cytology. We suggest that cell cannibalism is a dependable cytological feature of malignancy. PMID- 12561028 TI - Vitreous cytology in asteroid hyalosis and observations on interpretation of cytologic findings in specimens of the posterior (vitreous) compartment of the eye. AB - Cytologic methods have been used in the pathologic investigation of the posterior (vitreous) compartment of the eye in specimens obtained by fine-needle aspiration, as well as by surgical procedures. The vitreous body, which is in a semiliquid gel state, lends itself to pathologic investigation by cytologic methods. We report on a case of reparative eye surgery for complications of previous cataract extraction; vitrectomy was performed for vitreous opacities, which, by cytologic examination, were diagnostic of asteroid hyalosis, a relatively uncommon condition affecting the elderly. The cytologic findings diagnostic of this disease are described. Observations are made on the cytologic findings commonly encountered in specimens obtained from the posterior (vitreous) compartment of the eye. PMID- 12561029 TI - Fine-needle aspiration of adult rhabdomyoma: a case report with review of the literature. AB - The cytologic features of adult rhabdomyoma, a rare benign tumor of skeletal muscle origin, have been infrequently reported in the literature. We present here a case of a rhabdomyoma involving the floor of the mouth of a 78-year-old man initially diagnosed by fine-needle aspiration cytology. Cytologic features seen on Papanicolaou-stained smear preparation included cohesive clusters of skeletal muscle cells having abundant eosinophilic cytoplasm and often peripherally located nuclei. Although cross-striations and elongated intracytoplasmic inclusions were not identified in the smears, they were noted in the cell block preparation of the aspirated specimen and in the touch preparation and histologic sections of the surgically resected specimen. The cytologic differential diagnosis of this tumor is discussed. PMID- 12561030 TI - Diagnostic value of eosinophils in pleural effusion: a prospective study of 26 cases. AB - Eosinophilic pleural effusions (EPFs), defined as the presence of 10% or more eosinophils in the pleural fluid, are relatively rare. Their diagnostic and prognostic significance, however, remains controversial, as most of the studies are based almost entirely on retrospective case studies. This prospective study examines 26 eosinophilic pleural effusions from among 444 consecutive pleural effusions investigated at this tertiary health care center from October 1999 to April 2002. This study was attempted to unravel the diagnostic and prognostic significance of these eosinophilic effusions and assess their clinical implications, if any. Koss and Light's criteria were applied in the analysis, which comprised macroscopic, biochemical, cytological, and microbiological examinations. Of the 26 EPFs studied, five were associated with tuberculosis and three with metastatic disease. Nineteen patients had significant associated lymphocytosis. Twenty-four patients have been followed up and are in good health to date and have had no recurrence of effusion. Thus, EPF could be associated with inflammatory, benign, and malignant conditions. Hence, a closer search for a definite etiological agent is warranted in the setting of such an effusion, especially in populations endemic for tuberculosis, as in a developing country like India and in populations with a high prevalence of malignancy. PMID- 12561031 TI - Fine-needle aspiration biopsy of a cystic pleomorphic adenoma with extensive adnexa-like differentiation: differential diagnostic pitfall with mucoepidermoid carcinoma. AB - Although fine-needle aspiration biopsy (FNAB) is a highly accurate tool for the diagnosis of pleomorphic adenomas, even this common salivary gland neoplasm can be diagnostically challenging and cause pitfalls in cytodiagnosis. In particular, the presence of either cystic degeneration or squamous and mucinous metaplasia can lead to a false positive diagnosis of malignancy. Here we present the case of a 16-year-old female who presented with a painless, slowly growing mass in the superficial lobe of the right parotid gland. Magnetic resonance imaging of the parotid demonstrated a mass with heterogeneous postcontrast enhancement and a central, nonenhancing area suggestive of necrosis. FNAB of the lesion yielded proteinaceous debris and numerous whorls of keratin, small cohesive clusters of basaloid and squamoid epithelial cells, and many vacuolated and foamy cells. Initially and after consultation at an outside institution, the FNAB was reported as an "atypical neoplasm, cannot exclude mucoepidermoid carcinoma." A total, nerve-sparing parotidectomy and level II neck dissection revealed a pleomorphic adenoma with central cystic degeneration and extensive mixed appendageal differentiation. While foci of squamous metaplastic changes may occur in pleomorphic adenomas, the combination of cystic degeneration and extensive appendageal differentiation with numerous keratotic cysts is rare, and it presents the potential for misinterpretation of the FNAB as indicative of malignancy in general, and mucoepidermoid carcinoma in particular. PMID- 12561032 TI - Spontaneous infarction in fibroadenoma. PMID- 12561034 TI - Intraocular tuberculosis mimicking retinoblastoma: a case report. PMID- 12561035 TI - Solitary pulmonary nodule found incidentally in a smoker at Galveston, Texas. PMID- 12561036 TI - Cytopathology in Germany. PMID- 12561039 TI - IPL technology: a review. AB - BACKGROUND AND OBJECTIVES: Intense pulsed light (IPL) systems are high-intensity light sources, which emit polychromatic light. Unlike laser systems, these flashlamps work with noncoherent light in a broad wavelength spectrum of 515 1,200 nm. These properties allow for great variability in selecting individual treatment parameters and adapting to different types of skin types and indications. The purpose of this article was to critically review international medical publications of the many indication in which IPL technology can be used, including our own evaluations and experiences. STUDY DESIGN/MATERIALS AND METHODS: The range of therapeutic uses for high-intensity flashlamps was reviewed, ranging from benign cavernous hemangiomas, benign venous malformations, essential telangiectasias, leg telangiectasias, poikiloderma of Civatte, and port wine stains to pigmented lesions, cosmetically undesired hypertrichosis, and facial rhydids. The relative benefits and risks were discussed in detail and compared with other laser systems. RESULTS: Because of the wide spectrum of potential combinations of wavelengths, pulse durations, pulse frequency, and fluences, a great deal of experience is required when using IPL technology. Proper patient selection and critical diagnostics serve to keep the adverse effects of the treatment to a minimum. CONCLUSIONS: The distinctive technical conditions involved combine to make IPL technology an alternative and auxiliary treatment option to existing laser systems and conventional therapies. PMID- 12561040 TI - In vitro study examining the effect of sub-lethal QS 755 nm lasers on the expression of p16INK4a on melanoma cell lines. AB - BACKGROUND AND OBJECTIVES: Q-switched lasers had been used in the treatment of lentigo maligna but their role remains controversial. While previous studies have addressed the change in adhesion molecule expression after sub-lethal laser damage, no study has addressed the impact of sub-lethal laser damage at a molecular level. The p16 gene has been proposed as the candidate gene for melanoma. Our objective is to examine the effect of sub-lethal laser damage on p16 expression in melanoma cell lines. STUDY DESIGN/MATERIALS AND METHODS: Three human melanoma cell lines-HTB 66, Sk-mel-24 (HTB 71), and G361-were irradiated by a Q-switched 755 nm Alexandrite laser at fluencies that ranged from 0.85 to 2.0 J/cm(2). HTB 66 was the only cell line with significant expression of p16INK4a while the other two cells lines were p16INK4a negative and served as negative control. Protein and mRNA expression for p16 were assessed by flow cytometry and RT-PCR, respectively. RESULTS: The level of p16INK4a protein in cell line HTB 66 increased significantly after laser irradiation as compared with non-irradiated cells. The level of p16INK4a protein did not change in p16INK4a-negative cell lines (Sk-mel-24 and G361). However, there was only a slight increase in the percentage of G0/G1 phase cells. CONCLUSIONS: Sub-lethal laser damage could increase DNA damage leading to an increase in p16 expression, and such effect would be particularly undesirable for patients with p16 mutation. Further studies are warranted to examine the role of sub-lethal laser damage in inducing p16 mutation. PMID- 12561041 TI - Reduction in lateral thermal damage using heat-conducting templates: a comparison of continuous wave and pulsed CO2 lasers. AB - BACKGROUND AND OBJECTIVES: The advantages of the continuous wave (c.w.) CO(2) laser are offset by the delay in laser wound healing secondary to thermal damage. We have developed novel heat-conducting templates to reduce laser thermal damage. Because shortened pulse durations also decrease thermal damage, we tested the effectiveness of heat-conducting templates with a c.w. CO(2) clinical laser and a short-pulsed CO(2) laser to determine the best method and mechanism to minimize thermal damage. STUDY DESIGN/MATERIALS AND METHODS: Comparison of 0.2-second shuttered c.w. and 5-microsecond pulsed CO(2) lasers were made by doing incisions on 150 tissue samples from reduction mammoplasties and abdominoplasties. Copper, aluminum, glass, and Plexiglass heat-conducting templates were tested against no template (air) with both lasers. Histological samples were evaluated using computerized morphometrics analysis. RESULTS: Statistically significant reductions in lateral thermal damage were seen with the copper (50%) and aluminum (39%) templates used with the c.w. CO(2) laser. Only the copper template (39%) significantly reduced thermal damage when used with the pulsed CO(2) laser. Less thermal damage was seen using the pulsed CO(2) laser compared to the c.w. CO(2) laser with each template. CONCLUSIONS: Heat-conducting templates significantly reduced the amount of lateral thermal damage when used with the c.w. CO(2) laser (copper and aluminum) and short-pulsed CO(2) laser (copper). The c.w. CO(2) laser with the copper template compared favorably to the short-pulsed CO(2) laser without a template. Therefore, both heat conductive templates and short-pulse structure provide successful methods for reducing lateral thermal damage, and a combination of the two appears to provide optimal results. PMID- 12561042 TI - Topical rose bengal: pre-clinical evaluation of pharmacokinetics and safety. AB - BACKGROUND AND OBJECTIVES: Rose bengal (RB) is a potent photosensitizer that has largely been overlooked as a potential photodynamic therapy (PDT) agent. In this study, the feasibility of topical delivery of RB to the epidermis has been evaluated. STUDY DESIGN/MATERIALS AND METHODS: Topical formulations of RB were assessed on murine and rabbit skin for pharmacokinetic properties, cutaneous toxicity, and photosensitization. RESULTS: Hydrophilic formulations ( L(c), T(min) is dependent on deltat and L, while t(r) is a function of deltat only; (2) for spurts at L > L(c), T(min) still depends on L but not on deltat, while t(r) becomes a function of both deltat and L. Finally, for all deltat, t(r) reaches a maximum at L = 40 mm. CONCLUSIONS: Based on our results, a good choice to achieve low T(min) and t(r) for the treatment of superficial skin lesions may be met by using deltat of approximately 30-50 milliseconds and the shortest spray distance that is tolerable by the patient. Spurt durations (deltat) of more than 30-50 milliseconds at spray distances (L) greater than L(c) lead to higher T(min) and longer t(r). These parameters may be appropriate for laser therapy of deeper targets. PMID- 12561051 TI - Using a "non uniform pulse sequence" can improve selective coagulation with a Nd:YAG laser (1.06 microm) thanks to Met-hemoglobin absorption: a clinical study on blue leg veins. AB - BACKGROUND AND OBJECTIVES: Evaluation of the efficacy, on 1-2 mm blue leg telangiectasia, of a 1,064 nm Nd:YAG laser emitting in a non uniform pulse sequence calculated to consider Met-Hb formation during laser irradiation of a blood vessel. MATERIALS AND METHODS: A 1,064 nm Nd:YAG laser (Quantel Medical, Athos, France) was used in a non uniform pulse sequence mode, fluences: 300-360 J/cm(2) spot: 2 mm, + 5 degrees C contact cooling. The clinical evaluation was performed on 11 female patients, average age: 43 (25-57) years, phototype I-VI. All subjects were previously examined with Doppler ultrasound. A treatment site (6 x 4 cm) was selected on each patient. The topography of the vessels network was reported on a tracing plastic frame before each session and 6 weeks after the last one. These frames were digitized and the number of vessels was determined using the Digitized Tracing Frames Technique. Side effects were noted before and after every treatment, and 6 weeks after the last one. This study lasted for 10 months. RESULTS AND DISCUSSIONS: Patients tolerated the procedure without anesthesia. Moderate pain, transient erythema and edema, one hyperpigmentation and one matting were noted. There was no hypopigmentation. 55% (P < 0.002) vessels clearance after one session, 86% after two sessions (P < 0.001), and 98% (P < 0.001) after three sessions were obtained. On two patients, the treatment was completed after two sessions with a full clearance. Data reported in this study were obtained thanks to a computerized calculation of vessels clearance. They are similar or superior to those reported in the literature about 1,064 nm Nd:YAG lasers and leg telangiectasia. CONCLUSIONS: Since, it was developed to consider the modification of blood absorption and the methemoglobin formation which leads to an increase of the 1.06 microm wavelength absorption, the non uniform pulse mode emphasizes the efficacy of this 1,064 nm Nd:YAG laser concerning the treatment of blue leg veins telangiectasia between 1 and 2 mm. This mode gives the possibility to deliver high energy while preserving the surrounding tissue and leads to a rapid vessel clearance with reduced pain and few side effects when compared to previously published clinical studies using a 1.06 microm laser. PMID- 12561053 TI - Diagnostic evaluation of developmental delay/mental retardation: An overview. AB - Mental retardation (MR) is one of the few clinically important disorders for which the etiopathogenesis is still poorly understood. It is a condition of great concern for public health and society. MR is currently defined as a significant impairment of cognitive and adaptive functions, with onset before age 18 years. It may become evident during infancy or early childhood as developmental delay (DD), but it is best diagnosed during the school years. MR is estimated to occur in 1-10% of the population, and research on its etiology has always been a challenge in medicine. The etiopathogenesis encompasses so many different entities that the attending physician can sometimes feel a "virtual panic," starting a wide-range diagnostic evaluation. The Consensus Conference of the American College of Medical Genetics has recently established guidelines regarding the evaluation of patients with MR [Curry et al., 1997], emphasizing the high diagnostic utility of cytogenetic studies and neuroimaging in certain clinical settings. However, since then there has been substantial progress in molecular cytogenetics and neuroimaging techniques, the use of which has allowed recognition and definition of new disorders, thus increasing the diagnostic yield. This review will focus on the most appropriate investigations shown to be, at present, necessary to define the etiology of DD/MR, in the context of recommendations for the clinical evaluation of the patient with undiagnosed MR. PMID- 12561054 TI - Advances in molecular cytogenetics for the evaluation of mental retardation. AB - Recent years have witnessed rapid advances in molecular cytogenetics and its impact in studying mental retardation (MR). We review new molecular cytogenetic methods, including interphase fluorescence in situ hyrbridization (FISH), comparative genomic hybridization (CGH), multicolor karyotyping, telomere FISH, primed in situ labeling (PRINS), genotyping, microdissection, and microarray for the evaluation of MR. These new methods are very useful in two major aspects: further characterization of chromosome abnormalities as detected with routine banding analysis, including additions, duplications, deletions, translocations, markers, or complex aberrations; and screening for "hidden" chromosome aberrations in patients with an apparently normal karyotype. These new methods have great diagnostic potential in prenatal, postnatal, and preimplantational settings. Although powerful, at this point, they are primarily research tools in nature. It is essential that these new methods be used in conjunction with standard methods in order to maximize obtainable information for better management of patients with MR. PMID- 12561055 TI - Neuroimaging studies in the evaluation of developmental delay/mental retardation. AB - The employment of neuroimaging studies in the evaluation of individuals with developmental delay/mental retardation (DD/MR) is still highly debated. The Consensus Conference of the American College of Medical Genetics has suggested that "neuroimaging appears to have an especially important role in patients with microcephaly or macrocephaly, seizures, loss of psychomotor skills and neurologic signs," whereas the value of neuroimaging investigations "in the normocephalic patient without focal neurological signs is unclear" [Curry et al., 1997]. However, recent literature reports show how the latest neuroimaging techniques (in vivo proton magnetic resonance spectroscopy [H-MRS]) may prove to be useful in the diagnostic process of those individuals with DD/MR and no neurological signs/symptoms. The use of these techniques can, in addition, help in monitoring treatment in distinct metabolic disorders. This review will focus on the usefulness of neuroimaging studies in some of the newer metabolic disorders. This paper will also cover those recognizable patterns of human malformation where neuroimaging findings seem to be relevant both toward diagnosis and management, and add to our understanding of the related behavior phenotype. The essential role of magnetic resonance imaging (MRI) on the progress in the diagnostic recognition of malformations of cerebral cortical development is stressed. PMID- 12561056 TI - Metabolic disorders and mental retardation. AB - The metabolic and anatomical substrate of most forms of mental retardation is not known. Because the basis of normal brain function is not sufficiently understood, the basis of abnormal function is understood poorly. Even in disorders where the fundamental biochemical defect is known, such as phenylketonuria (PKU) and other enzyme defects, the exact basis for brain dysfunction is uncertain. The outcome for treated PKU, galactosemia, homocystinuria, and lysosomal disorders is not yet optimal. The various forms of nonketotic hyperglycinemia often respond poorly to current therapy. Less familiar disorders, with or without seizures, such as deficient synthesis of serine or creatine and impaired glucose transport into the brain, and disorders with variable malformations, such as Smith-Lemli-Opitz (SLO) syndrome and the congenital disorders of glycosylation (CDGs), may initially be thought to be a nonspecific form of developmental delay. Less familiar disorders, with or without seizures and disorders with variable malformations may initially be thought to be a nonspecific form of developmental delay. Simple tests of urine, blood, and cerebrospinal fluid may lead to a diagnosis, accurate genetic counseling, and better treatment. Metabolic brain imaging (magnetic resonance spectroscopy (MRS)) has also helped to reveal biochemical abnormalities within the brain. PMID- 12561057 TI - Costello syndrome: an overview. AB - The Costello syndrome is characterized by prenatally increased growth, postnatal growth retardation, coarse face, loose skin resembling cutis laxa, nonprogressive cardiomyopathy, developmental delay, and a outgoing, friendly behavior. Patients can develop papillomata, especially around the mouth, and have a predisposition for malignancies (mainly abdominal and pelvic rhabdomyosarcoma in childhood). Costello syndrome is likely to be an autosomal dominant disorder. The pathogenesis is unclear, but there are many clues for a disturbed elastogenesis, possibly through a disturbed elastin-binding protein reuse by chondroitin sulfate bearing proteoglycans accumulation. A review of the findings in the 73 patients that have been described in sufficient detail is provided. PMID- 12561058 TI - Mental deficiency, alterations in performance, and CNS abnormalities in overgrowth syndromes. AB - Mental deficiency, alterations in performance, and central nervous system (CNS) abnormalities are discussed in the following overgrowth syndromes: Sotos syndrome, Weaver syndrome, Proteus syndrome, neurofibromatosis type 1, fragile X syndrome, syndromes with neonatal hypoglycemia, Simpson-Golabi-Behmel syndrome, hemihyperplasia, Sturge-Weber syndrome, Bannayan-Riley-Ruvalcaba/Cowden syndrome, macrocephaly-autism syndrome, PEHO syndrome, chromosomal syndromes, and other miscellaneous syndromes. PMID- 12561059 TI - Kabuki make-up syndrome: a review. AB - Kabuki make-up syndrome (KMS, OMIM 147920) is an MCA/MR syndrome of unknown cause. It is characterized by a dysmorphic face, postnatal growth retardation, skeletal abnormalities, mental retardation, and unusual dermatoglyphic patterns. Approximately more than 350 cases have been reported from all over the world. Besides these five cardinal manifestations, joint laxity (74%), dental abnormalities (68%), and susceptibility to infections including recurrent otitis media (63%) were well recognized as other frequent features. A variety of visceral anomalies such as cardiovascular anomalies (42%), renal and/or urinary tract anomalies (28%), biliary atresia, diaphragmatic hernia, and anorectal anomaly were also reported. Some patients were said to have normal intelligence (16%) and normal heights, suggesting that they may have reproductive fitness to have their children. At least eight patients had lower lip pits with or without cleft palate, known as a feature of van der Woude syndrome. There have been 13 chromosomal abnormalities associated with KMS. However, no common abnormalities or breakpoints that possibly contribute to positional cloning of the putative KMS gene(s) are known. Although clinical manifestations of KMS are well established, its natural history, useful for genetic counseling, remains to be studied. PMID- 12561060 TI - Genetics, mental retardation, and the forging of new alliances. AB - The lives of people with intellectual disabilities are tightly intermeshed with the philosophies and practices of special educators, therapists, psychologists, social workers, and others who make up the mental retardation field. These nonmedical workers represent a kind of extended family, which can strongly influence the decisions made and services received by the people they serve. By contrast, most individuals with mental retardation do not have extraordinary medical needs, and healthcare professionals play only a minor role in their human services family. There are deeply rooted historical differences between the medical model of mental retardation with which genetics professionals are familiar and the philosophies embraced by nonmedical workers who dominate the mental retardation field. These contrasts have an important impact on the genetic diagnostic process, as well as a more global effect on mental retardation research. Without the full participation of teachers, therapists, psychologists, and others outside the medical setting, many practical applications of genetic research can neither be implemented nor expanded upon, and an important opportunity for cross-fertilization is lost. It is in the interest of the genetics community to take advantage of recent overtures by the mental retardation field to forge new alliances and broaden its perspectives on intellectual disability. PMID- 12561061 TI - Will we ever succeed in resolving the adjuvant treatment dilemma in rectal cancer? PMID- 12561062 TI - Association between extent of axillary lymph node dissection and patient, tumor, surgeon, and hospital factors in patients with early breast cancer. AB - BACKGROUND AND OBJECTIVES: Axillary lymph node dissection (ALND) in patients with breast cancer is crucial for accurate staging, provides excellent regional tumor control, and is included in the standard of care for the surgical treatment of breast cancer. However, the extent of ALND varies, and the extent of dissection and the number of lymph nodes that comprise an optimal axillary dissection are under debate. Despite conflicting evidence, several studies have shown that improved survival is correlated with more lymph nodes removed in both node negative and node-positive patients. The purpose of this study is to determine which patient, tumor, surgeon, and hospital characteristics are associated with the number of nodes excised in early breast cancer patients. METHODS: A random sample of 938 women with node-negative breast cancer was drawn from the Ontario Cancer Registry and the data supplemented with chart reviews. The extent of axillary dissection was studied by examining the number of nodes examined in relation to the patient, tumor, surgeon, and hospital factors. RESULTS: The mean number of lymph nodes excised was 9.8 (SD = 4.8; range, 1-31), and 49% of patients had >/=10 nodes excised. Lower patient age was associated with the excision of more lymph nodes (>/=10 nodes: 63% of patients <40 years vs. 38% of patients >/=80 years). Surgeon academic affiliation and surgery in a teaching hospital were highly correlated with each other and were significantly associated with the excision of >/=10 nodes. The number of nodes excised was not associated with any tumor factors, nor with the breast operation performed. These results were confirmed with multivariable models. CONCLUSIONS: Even though the number of lymph nodes found in the pathologic specimen can be influenced by factors other than surgical technique (e.g., number of nodes present, specimen handling, and pathologic examination), this study shows significant variation of this variable and an association with several patient and surgeon/hospital factors. This variation and the association with survival warrant further study and effort at greater consistency. PMID- 12561063 TI - Intradermal radioisotope injection is superior to subdermal injection for the identification of the sentinel node in breast cancer patients. AB - BACKGROUND AND OBJECTIVES: The purpose of the present study was to evaluate whether the intradermal injection of radiocolloids would improve the identification rate of sentinel nodes over the subdermal injection in breast cancer patients. METHODS: Sentinel node biopsy was performed in T2 breast cancer patients with clinically negative nodes, using subdermal or intradermal injection of radioisotopes with the peritumoral dye injection. We used Tc-99m tin colloid, with a larger particle size (0.4-5 microm), rather than sulfur colloid and colloidal albumin. RESULTS: The initial 55 patients underwent subdermal injection of radiocolloids; the next 61 patients underwent intradermal injection of radiocolloids for sentinel node biopsy. The detection rate of sentinel nodes was significantly (P = 0.048) higher in the intradermal injection group (61/61, 100%) than in the subdermal injection group (51/55, 92.7%). False-negative rates were comparable between the two groups. Lymphoscintigraphy visualized the sentinel nodes significantly (P < 0.0001) more often in the intradermal injection group (59/61, 96.7%) than in the subdermal injection group (20/54, 37.0%). CONCLUSIONS: A significantly higher identification rate of sentinel node biopsy and lymphoscintigraphy can be achieved by intradermal injection of Tc-99m tin colloid with a large particle size than by subdermal injection. PMID- 12561065 TI - Significance of reduction surgery in multidisciplinary treatment of advanced hepatocellular carcinoma with multiple intrahepatic lesions. AB - BACKGROUND AND OBJECTIVES: By comparing the survival rates of patients treated with or without surgery, the significance of, and the indication for, reduction surgery in the multidisciplinary treatment of patients with HCC with multiple intrahepatic lesions were examined. METHODS: In patients with HCC with multiple intrahepatic lesions, cumulative survival rates were determined and compared for 28 patients (group S) who underwent reductive hepatic resection and 43 (group N) who were treated nonsurgically by transcatheter arterial infusion chemotherapy (TAI), transcatheter arterial chemoembolization (TACE), or percutaneous transhepatic ethanol injection therapy. In group S, 20 patients had adjuvant therapy, consisting of ethanol injection therapy or microwave coagulonecrotic therapy for the remaining satellite lesions during hepatectomy, and all patients in this group underwent TAI or TACE postoperatively. The influence of surgery on patient survival was examined by multiple regression analysis using the Cox's hazard model; then, for each prognostic factor, survival rates were obtained and compared between the groups. RESULTS: In group S, the 1-, 3-, and 5-year cumulative survival rates were 58.2%, 27.1%, and 21.7%, whereas the corresponding values in group N were 34.3%, 4.7%, and 4.7%, the difference being statistically significant (P = 0.0239). In group S, the 1-, 3-, and 5-year cumulative survival rates for patients without intraoperative adjuvant therapy were 25%, 0%, and 0%, whereas those for patients with intraoperative adjuvant therapy were 72.7%, 41.3%, and 33.0% (P = 0.001). Multiple regression analysis showed that hepatic resection, the Child-Pugh score, and the size of the main tumor affected survival independently. Univariate analysis of differences in the cumulative survival rates between the groups as a function of prognostic factor showed that group S had statistically significant better survival rates than group N in those subgroups of patients who were <60 years old, with HBV infection, with a Child Pugh score of 5 or 6, with a main tumor of <5-cm diameter, with <5 tumors, or without portal thrombi. CONCLUSIONS: When combined with intraoperative adjuvant therapy for remaining satellite tumors, reduction surgery provided survival benefit for patients with HCC with multiple intrahepatic lesions in those groups of patients selected by criteria determined in this study. PMID- 12561066 TI - Prognostic use of growth characteristics of early gastric cancer and expression patterns of apoptotic, cell proliferation, and cell adhesion proteins. AB - BACKGROUND AND OBJECTIVES: Selection of suitable treatment for early gastric cancers, such as endoscopic mucosal resection or the major surgical option of resection of the cancer together with a radical lymph node dissection, may be assisted by comparing the growth characteristics of the cancer with selected molecular characteristics. The results could be used to predict those cases that have a higher risk of developing secondary metastases. METHODS: A total of 1,196 Japanese patients with early gastric cancers (648 mucosal cancers and 548 submucosal) were included in the selection of two groups: a metastatic group made up 57 cancers with lymph node metastasis (9 mucosal, 48 submucosal), and a nonmetastatic group of 61 cases (6 mucosal, 55 submucosal) without lymph node metastasis. Growth characteristics of the cancers (superficially spreading, penetrating or invasive, lymph node metastasis) were compared with immunohistochemical expression of single-stranded DNA (ssDNA) protein (apoptosis indicator), bcl-2 and p53 (apoptosis-associated), Ki-67 (cell proliferation), and E-cadherin (cell adhesion) proteins. RESULTS: The lesions in the nonmetastatic group had higher levels of apoptosis and lower expression of bcl-2 than in the metastatic group, indicating an inhibitory role for apoptosis in malignant progression. Apoptosis was also higher in the superficial compared with the invasive lesions of both groups. The lesions in the metastatic group had higher p53 expression than that of the nonmetastatic group, whereas apoptosis in the metastatic group was lower than in the nonmetastatic group. An unproved explanation for this finding may be that, although increased, p53 was mutated and ineffective in promoting apoptotic control of metastatic progression. E-cadherin was decreased in the invasive lesions of both groups, indicating a greater ability of these cells to lose adhesion, to invade the submucosa, and to metastasize. Cell proliferation was highest in the superficial lesions of both metastatic and nonmetastatic groups. CONCLUSIONS: Early gastric cancers with low levels of apoptosis, increased bcl-2, and high levels of p53 expression are more likely to invade and metastasize. PMID- 12561067 TI - p53 alteration is not an independent prognostic indicator, but affects the efficacy of adjuvant chemotherapy in human pancreatic cancer. AB - BACKGROUND AND OBJECTIVES: Mutations in the p53 gene are found in more than 50% of human cancers and are observed in 60-80% of pancreatic cancers. The clinicopathologic implications of p53 abnormalities and their effects on the efficacy of the adjuvant chemotherapy for pancreatic cancer remain controversial. METHODS: We investigated the p53 status in core exon-4 to -9 (codon 33-331) by direct DNA sequencing in a series of 72 pancreatic cancers and analyzed the effects of p53 abnormalities on the patients' survival and the efficacy of adjuvant chemotherapy. RESULTS: p53 mutations were found in 62.5% (45/72) of cases, including 38 point mutations and 7 frameshift mutations. The subtypes of p53 mutations included 68.9% (31/45) transitions and 15.6% (7/45) transversions. 39.5% (15/38) of point mutations were CGT (Arg) to CAT (His) mutation at codon 273 of exon-8. 34.2% (13/38) of point mutations were CGG (Arg) to TGG (Trp) mutation at codon-248 of exon-7. Of seven frameshift mutations, four were seen at exon-4, two at exon-5, and one at exon-6. Of overall cases, p53 abnormalities were not associated with a poorly differentiated grade and an advanced stage. The relationship of adjuvant chemotherapy to survival is approaching statistical significance. Univariate analysis showed that in the p53 mutation group, the patients who received adjuvant chemotherapy had a better survival ratio than that of patients who did not do. Multivariate analysis indicated that in the group with p53 mutations, the significant factors for survival were adjuvant chemotherapy, histologic grade, and clinical stage. However, in the group with a wild-type p53 gene, only histologic grade was a significant factor. In addition, 34.7% (25/72) of the cases harbor p53 polymorphism mutation only at codon-72 of exon-4, which did not show any significant effect on the pathology, prognosis, and efficacy of adjuvant chemotherapy of the pancreatic cancers. CONCLUSIONS: A p53 abnormality was not an independent factor for evaluating the prognosis of patients with pancreatic cancer, but was a beneficial indicator for selecting a reasonable strategy of adjuvant chemotherapy against pancreatic cancer. PMID- 12561068 TI - Establishment of a nude mouse model of hepatic metastasis for evaluation of targeted retroviral gene delivery. AB - BACKGROUND AND OBJECTIVES: The goal of this study was to establish a reproducible nude mouse model of liver metastasis and investigate the potential of deploying targeted injectable retroviral vectors for metastatic gastrointestinal cancer. METHODS: Human cancer cells were injected into the portal vein via an indwelling catheter. The animals were sacrificed at specified time intervals, and the number of tumor nodules was counted in histologic sections of harvested livers. A group of animals received either an extracellular matrix-targeted or a nontargeted retroviral vector bearing a beta-galactosidase gene by portal vein infusion. RESULTS: The number of tumor nodules increased progressively over time at 150 min. and dialysis to be independent risk factors for hospital death. Actuarial survival at 7.5 years overall was 39% in the 80-plus age group vs. 53% in the 70 79 age group for VHD and 38% in the 80-plus age group vs. 62% in the 70-79 age group. No significant difference was seen in survival between groups for IHD. Stroke proved to be an independent prognostic factor. CONCLUSIONS: Cardiac surgery is conducted feasibly in selected octogenarians, providing acceptable mortality and results similar to those achieved in those 70 to 79 years old. PMID- 12561092 TI - Minimally invasive diagnosis and treatment of traumatic rupture of the right hemidiaphragm with liver herniation. AB - We report a case of blunt traumatic rupture of the right hemidiaphragm with liver herniation. A 57-year-old man admitted in an emergency after a traffic accident was suspected from chest radiography and computed tomography to have traumatic diaphragmatic rupture. Magnetic resonance imaging was helpful in the final diagnosis. Thoracoscopy was useful in planning surgery and surgically repairing the ruptured diaphragm. PMID- 12561093 TI - Chylopericardium following mitral valve replacement. AB - A 49-year-old man with mitral regurgitation developed chylopericardium after mitral valve replacement. On postoperative day (POD) 1, the patient was started on a regular diet. On POD 2, drainage from retrosternal and intrapericardial tubes increased, becoming turbid and milky. Because the triglyceride concentration was high at 197 mg/dl, we diagnosed the condition as chylopericardium. The patient was given nothing by mouth for 2 days and, from POD 4, received a low-fat diet, after which drainage cleared and volume decreased. The 2 drainage tubes were removed on POD 6, and he was discharged without complications on POD 18. PMID- 12561094 TI - Thoracic aortic aneurysm associated with pseudocoarctation of the aorta. AB - Pseudocoarctation is a rare anomaly in the descending thoracic aorta. A 44-year old man experienced sudden onset of back pain for 5 days prior to admission. Computed tomography showed kinking and stenosis in the distal aortic arch with a distal aneurysm. The patient underwent emergency surgery, with a diagnosis of impending rupture. The aneurysm was lobular with a very thin wall. Pseudocoarctation is rare and most often is asymptomatic. However, the aneurysm should be treated surgically, and the area of stenosis resected. PMID- 12561095 TI - Coronary artery bypass grafting to a calcified right coronary artery. AB - A severely calcified coronary artery demands a special technique in coronary artery bypass surgery. We have successfully developed a "punch-out" technique for a calcified right coronary artery in a dialysis patient. After an incision into the target coronary artery, the calcified arterial wall was resected using a punch to make an oval hole for anastomosis. Limited endarterectomy, which consisted of dissection and removal of the calcified endothelium and media around this hole, preserving the adventitia, enabled the anastomosis. This technique can be used for a severely calcified coronary artery that is unamendable to conventional coronary artery bypass grafting. PMID- 12561096 TI - Off-pump coronary artery bypass grafting in a patient with liver cirrhosis. AB - We report a case of unstable angina pectoris and alcohol-related Child-Pugh class B cirrhosis. The patient was a 60-year-old man who was admitted to hospital with chest pain. He had previously been diagnosed to have Child B cirrhosis due to alcoholic liver dysfunction at 58 years of age. He also had experienced ruptured esophageal varices, moderate ascites, and hyperammonemia. We performed percutaneous catheter intervention; however, he developed re-stenosis in the right coronary artery, and progression in the disease in other coronary arteries. We then performed coronary artery bypass grafting on the beating heart without cardiopulmonary bypass. He was discharged on the 13th postoperative day without any complications. This case demonstrated that off-pump coronary artery bypass grafting was safe for such a patient. PMID- 12561097 TI - Paravalvular leakage causing hemolytic anemia at 8-9 years after mitral valve replacement. AB - We report two similar cases of women, aged 52 and 71 years, who developed shortness of breath or easy fatigability at 8-9 years after mitral valve replacement of the mitral valve in each patient, the echocardiographic study revealed paravalvular leakage. Previous examination before the appearance of symptoms had revealed a well-functioned prosthetic valve, and no hematological abnormality had been detected. The hemoglobin levels at admission were 6.4 and 6.6 g/dl, and their serum lactate dehydrogenase levels were 5218 and 4608 mU/ml. Each successfully underwent another valve replacement, and after surgery, the hemolytic anemia and the symptoms disappeared. PMID- 12561098 TI - Unsuspected primary pulmonary malignant lymphoma. AB - A 70-year-old man referred for treatment of a left lower lung tumor was shown in chest computed tomography to have a homogeneous round tumor 45 mm in diameter with an enhanced thin wall in the lower lobe of the left lung. No specific finding was seen in material obtained by transbronchial and computed tomography (CT)-guided lung tissue biopsy, so the presumptive diagnosis was a lung abscess. Despite antibiotics administered for 2 weeks, radiography showed the tumor had grown, necessitating left lower lobectomy. The permanent section was diagnosed as diffuse large B-cell lymphoma. Because CT findings for the tumor suggested a lung abscess and the central part of the tumor consisted of fibrotic and necrotic tissue, we had difficulty establishing a final diagnosis. The literature showed primary pulmonary lymphomas yielded a variety of findings radiographically, making surgery paramount for ascertaining a final diagnosis. PMID- 12561099 TI - Adenosquamous carcinoma arising in Barrett's esophagus. AB - Primary adenocarcinoma of the esophagus is rare in Japan and, in most cases, arises from Barrett's esophagus epithelium. A 72-year-old man reporting heartburn and dysphagia and preoperatively diagnosed with adenosquamous carcinoma arising from Barrett's esophagus underwent thoracic esophagectomy and lymph node dissection in curative resection. Pathological diagnosis of the resected specimen showed adenosquamous carcinoma (coexistent adenocarcinoma and squamous cell carcinoma) invasive to the submucosal layer; metastasis was found in regional lymph nodes. Pathological staging was pT1bN1M0, stage II. Unfortunately, the man died of liver and lung metastasis 17 months postoperatively. To our knowledge, this rare case is only the fifth reported in the English literature on adenosquamous carcinoma arising from Barrett's esophagus. PMID- 12561100 TI - Coronary revascularization in the 21st century. Emphasis on contributions by Japanese surgeons. AB - The first three decades of coronary artery surgery have provided the foundation for the next century of this evolution. It is apparent that a multitude of events including the development of cardioplegia, improving surgical instrumentation, technological advances including endoscopic approaches and computer assisted robotics and biologic discoveries such as the role of the endothelium have provided the underpinnings for improved surgical outcomes. However, the single most important determinant of late results is the type of bypass conduit used for grafting. Thus, use of the left internal thoracic artery (ITA) grafted to the left anterior descending coronary is a more important determinant of survival than is any other factor (progression of coronary artery disease, increased age, poor left ventricular function, diabetes, female gender and off-pump operations). Use of two ITAs provides further benefit and it is likely that three or more arterial conduits will be shown to be advantageous in this regard in due time. Japanese cardiothoracic surgeons have made significant contributions to the continuing evolution of coronary bypass surgery and particularly to the advance of arterial conduits. This report will address those contributions to this evolution. PMID- 12561101 TI - Markov-type evolution of materials into a polar state. AB - Assembling polar building blocks into a solid material by a Markov-chain process of unidirectional growth principally results in a metastable state that shows effects of macroscopic polarity. Stochastic polarity formation can be described by probabilities for the attachment of building blocks to a surface. Because of the polar symmetry of the building blocks, there is a fundamental difference in the probabilities for attaching them "tip-first" or "back-first" to growth sites at a surface. A difference in the corresponding probabilities drives the evolution of a vectorial property through a gain in configurational entropy. Examples from the mechanical, the crystalline and the biological world demonstrate growth-induced macroscopic polarity. In crystals, growth upon centrosymmetric seeds can produce twinned crystals with a "sectorwise" pyroelectric effect. Polarity formation in connective tissues is explained by a Markov-chain mechanism, which drives the self-assembly of collagen fibril segments. An unified stochastic growth model brings up a general concept for the formation of materials with polar properties. PMID- 12561102 TI - New transition-metal-dependent DNAzymes as efficient endonucleases and as selective metal biosensors. AB - Like proteins and RNA molecules, many DNA molecules have now been shown to catalyze a variety of reactions and are thus called DNAzymes. With limited building blocks, DNAzymes need to recruit other cofactors in order to match other enzymes in terms of reaction diversity and catalytic efficiency. Several unique properties make transition-metal ions an ideal choice of cofactor for DNAzymes. Indeed, new DNAzymes that bind transition-metal ions with high affinity and selectivity have been obtained through the use of a powerful combinatorial biology tool called in vitro selection. This accomplishment now makes it possible to obtain different classes of metallo-DNAzymes in the laboratory within a short period of time. It also offers a rare opportunity to compare and contrast structural and functional properties of metal-binding sites in proteins and in DNAzymes. The resulting transition-metal-dependent DNAzymes have displayed high activity toward cleavage of DNA and RNA and thus hold promise for their biochemical and pharmaceutical applications. Finally, the use of DNAzymes as a new class of highly sensitive and selective biosensors for metal ions has been demonstrated recently. PMID- 12561103 TI - Second-generation mimics of ganglioside GM1 oligosaccharide: a three-dimensional view of their interactions with bacterial enterotoxins by NMR and computational methods. AB - As a step to delineate a strategy of ligand design for cholera toxin (CT), NMR studies were performed on several mimics of the GM1 ganglioside oligosaccharide. The conformation of these analogues was investigated first in solution and then upon binding to cholera toxin by transferred nuclear Overhauser effect (TR-NOE) measurements. It was demonstrated that CT selects a conformation similar to the global minima of the free saccharides from the ensemble of presented conformations. No evidence of major conformational distortions was obtained, but one or two of the available conformers of the hydroxyacid side chain appear to be selected in the bound state. The NMR data were interpreted with the aid of computer models, generated and analyzed by using a combination of different approaches (MacroModels' MC/EM and MC/SD, Autodock, and GRID). Analysis of the NMR data supported by computational studies allowed us to interpret the experimental observations and to derive workable models of the ligand:toxin complexes. These models suggest that the higher affinity of the (R)-lactic acid derivative 3 may stem from lipophilic interactions with a hydrophobic area in the toxin binding site located in the vicinity of the sialic acid side chain binding region of the CT:GM1 complex, and formed by the side chain of Ile-58 and Lys-34. Thus, the models obtained have allowed us to make useful design suggestions for the improvement of ligand affinity. PMID- 12561104 TI - 2,8'-disubstituted-1,1'-binaphthyls: a new pattern in chiral ligands. AB - The title binaphthyls 19 and 26, which are the positional isomers of 2-methoxy-2' (diphenylphosphino)-1,1'-binaphthyl (MOP, 19) and 2-amino-2'-hydroxy-1,1' binaphthyl (NOBIN, 26), have been synthesized by Suzuki coupling as the key step (10 + 15-->18), followed by functional group transformations, involving C-P and C N bond formation (18-->19 and 18-->23). Racemic intermediate 22 was resolved by co-crystallization with N-benzylcinchonidinium chloride and the absolute configuration determined by X-ray crystallography. These novel binaphthyls are configurationally stable and, as such, potentially usable as chiral ligands in asymmetric reactions. Michael addition of the glycine-derived enolate 40 to methyl acrylate, carried out in the presence of (R)-(-)-27 as the chiral phase transfer catalyst, afforded L-glutamic acid (S)-(+)-43 of 92% ee (after hydrolysis of the primary product). PMID- 12561105 TI - Synthesis of stable analogues of thiamine di- and triphosphate as tools for probing a new phosphorylation pathway. AB - Thiamine (vitamin B1) is an essential nutritional factor metabolized inside the body in its mono-, di-, and triphosphate forms. Although the action of thiamine and thiamine diphosphate have been intensely investigated, many questions remain unanswered and the role of thiamine triphosphate is still especially unknown. To probe recent hypotheses on the implication of thiamine triphosphate in a new phosphorylation pathway involving synaptic proteins, we synthesized a series of thiamine di- and triphosphate analogues that are resistant to both enzymatic and chemical hydrolyses. The key step in the preparation of the title compounds is the coupling of thiamine propyl disulfide with adequately protected methylenebis phosphonic acid, the corresponding triphosphate analogue, and difluoromethylenebisphosphonic acid. PMID- 12561106 TI - Engineering a remarkably low HOMO-LUMO gap by covalent linkage of a strong pi donor and a pi-acceptor--tetrathiafulvalene-sigma-polynitrofluorene diads: their amphoteric redox behavior, electron transfer and spectroscopic properties. AB - Novel R3TTF-sigma-A compounds 14, 16 and 19 (R3TTF = trial-kyletrathiafulvalene, sigma = saturated spacer, A = polynitrofluoren-9-dicyano-methylene acceptor) incorporating very strong donor and acceptor moieties have been synthesized by condensation of the corresponding R3TTF-sigma-fluoren-9-one diads with malononitrile. Reversible five-step amphoteric redox behavior has been observed with an extremely low HOMO-LUMO gap (approximately 0.3 eV). For compound 14 a strong EPR signal is observed in the solid state, ascribed to intermolecular complexation: a less intense signal is seen in solution, corresponding to ca. 2% of the molecules existing in a radical form at room temperature. Intramolecular charge transfer in diads 14 and 16 is manifested in strong absorption bands in the near-IR region of their electronic spectra. Spectroelectrochemical data reveal marked electrochromic behavior in the visible and near-IR region of both compounds. The first X-ray crystal structure of a fluorene radical-anion salt is reported, namely the copper salt of 2,4,5,7-tetranitro-9- dicyanomethylenefluorene (1:1 stoichiometry). PMID- 12561107 TI - Two-point self-coordination of a dizinc(II) bispyridylporphyrin ruthenium complex leading selectively to a discrete molecular assembly: solution and solid-state characterization. AB - The dizinc(II) bispyridylporphyrin ruthenium complex trans,cis,cis[RuCl2(CO)2(Zn.4'-cisDPyP)2] (1Zn, 4'-cisDPyP = 5,10-bis(4'-pyridyl) 15,20-diphenylporphyrin) features two donor (the uncoordinated 4'-N(py) atoms) and two acceptor (the Zn atoms) sites and is thus a building block suited for two point coordination. 1H NMR spectroscopy indicates that 1Zn self-assembles in solution through Zn-4'-N(py) interactions to yield selectively a highly symmetrical discrete species, in which all donor and all acceptor sites of 1Zn are mutually saturated. Single-crystal X-ray analysis established that this adduct is a dimeric species, (1Zn)2, with a global S4 symmetry, in which the four porphyrins have a propeller-like arrangement. The dimeric species (1Zn)2 is a meso form derived from the combination of two 1Zn units with opposite helical chirality. The geometry of this highly symmetrical tetraporphyrin assembly in solution, as determined by NMR spectroscopy, is essentially the same as that found in the solid state. Thus 1Zn is an unprecedented example of metal containing self-complementary building block that selectively recognizes itself through four Zn-N(py) interactions, and thus yields a very stable and symmetrical dimeric species, (1Zn)2, that features four porphyrins and six metal atoms (two Ru and four Zn). PMID- 12561108 TI - The trifluoromethoxy carbonyl peroxy radical CF3OC(O)OO. AB - CF3OC(O)OO radicals are generated by low-pressure flash thermolysis of CF3OC(O)OOOC(O)OCF3 highly diluted in inert gases and followed by subsequent isolation in an inert-gas matrix at low temperatures. The by-products CO2, COF2, CF3O, and CF3OO are detected. The new peroxy radical is characterized by IR and UV spectroscopy and by its UV photolytic decay which leads to the formation of CF3OO and CO2. According to DFT calculations the exitence of three stable rotamers is predicted and two of them are found experimentally. PMID- 12561109 TI - N1 and N3 linkage isomers of neutral and deprotonated cytosine with trans [(CH3NH2)2PtII. AB - A series of complexes obtained from the reaction of trans-[(CH3NH2)2PtII] with unsubstituted cytosine (CH) and its anion (C), respectively, has been prepared and isolated or detected in solution: trans-[Pt(CH3NH2)2(CH-N3)Cl]Cl.H2O (1), trans-[Pt(CH3NH2)2(CH-N3)2](ClO4)2 (1a), trans-[Pt(CH3NH2)2(C-N3)2].2H2O (1b), trans-[Pt(CH3NH2)2(CH-N3)2](ClO4)(2).2DMSO (1c), trans-[Pt(CH3NH2)2(CH-N1)2] (NO3)(2).3H2O (2a), trans-[Pt(CH3NH2)2(C-N1)2].2H2O (2b), trans-[Pt(CH3NH2)2(CH N1)(CH-N3)](ClO4)2 (3a), trans-[Pt(CH3NH2)2(C-N1)(C-N3)] (3b), and trans [Pt(CH3NH2)2(N1-CN3)(N3-C-N1)Cu(OH)]ClO(4).1.2H2O (4). X-ray crystal structures of all these compounds, except 3a and 3b, are reported. Complex 2a is of particular interest in that it contains the rarer of the two 2-oxo-4-amino tautomer forms of cytosine, namely that with the N3 position protonated. Since the effect of PtII on the geometry of the nucleobase is minimal, bond lengths and angles of CH in 2a reflect, to a first approximation, those of the free rare tautomer. Compared to the preferred 2-oxo-4-amino tautomer (N1 site protonated) of CH, the rare tautomer in 2a differs particularly in internal ring angles (7-11 sigma). Formation of compounds containing the rare CH tautomers on a preparative scale can be achieved by a detour (reaction of PtII with the cytosine anion, followed by cytosine reprotonation) or by linkage isomerization (N3-->N1) under alkaline reaction conditions. Surprisingly, in water and over a wide pH range, N1 linkage isomers (3a, 2a) form in considerably higher amounts than can be expected on the basis of the tautomer equilibrium. This is particularly true for the pH range in which the cytosine is present as a neutral species and implies that complexation of the minor tautomer is considerably promoted. Deprotonation of the rare CH tautomers in 2a occurs with pKa values of 6.07 +/- 0.18 (1 sigma) and 7.09 +/- 0.11 (1 sigma). This value compares with pKa 9.06 +/- 0.09 (1 sigma) (average of both ligands) in 1a. PMID- 12561110 TI - Structures, bond energies, heats of formation, and quantitative bonding analysis of main-group metallocenes [E(Cp)2] (E = Be-Ba, Zn, Si-Pb) and [E(Cp)] (E = Li Cs, B-Tl). AB - The geometries, metal-ligand bond dissociation energies, and heats of formation of twenty sandwich and half-sandwich complexes of the main-group elements of Groups 1, 2, 13, and 14, and Zn have been calculated with quantum chemical methods. The geometries of the [E(Cp)] and [E(Cp)2] complexes were optimized using density functional theory at the BP86 level with valence basis sets, which have DZP and TZP quality. Improved energy values have been obtained by using coupled-cluster theory at the CCSD(T) level. The nature of the metal-ligand bonding has been analyzed with an energy-partitioning method. The results give quantitative information about the strength of the covalent and electrostatic interactions between En+ and (Cp-)n (n = 1, 2). The contributions of the orbitals with different symmetry to the covalent bonding are also given. PMID- 12561111 TI - Selective and shape-selective Baeyer-Villiger oxidations of aromatic aldehydes and cyclic ketones with Sn-beta zeolites and H2O2. AB - Sn-Beta is used as a heterogeneous catalyst for the Baeyer-Villiger reaction with hydrogen peroxide. Cyclic ketones are transformed into the corresponding lactones, while unsaturated ketones are oxidized to the corresponding unsaturated lactones with very high chemoselectivity. The catalyst is also selective for the oxidation of aromatic aldehydes with H2O2, producing the formate ester or the corresponding hydrolyzed product, that is the alcohol. Shape-selective oxidations are observed for isomeric reactants with different molecular shapes. The catalytic Sn sites have been characterized by 119Sn MAS-NMR spectroscopy, and tetrahedral incorporation into the zeolite framework has been demonstrated. In situ IR spectroscopy and 18O labeling experiments have shown that the oxidation mechanism involves an intermediate of the Criegee type. PMID- 12561113 TI - Selective synthesis of fused cyclooctatetraenes by [4 + 4] coupling between two different diene units. AB - In the presence of CuCl and 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone, the [4 + 4] coupling between zirconacyclopentadienes and 1,3-diiodobutadienes fused through an oxygen or nitrogen five-membered ring proceeded at ambient temperature to afford fully substituted polycyclic cyclo-octatetraenes in good yields. The fused ring moiety of the diiodides plays a critical role. The corresponding acyclic diiodide and a cyclohexane-fused analogue gave no coupling product, and a cyclopentane derivative showed only moderate reactivity. Correlation of the structures of the diiodides and their reactivity was established by an X-ray and density functional study. PMID- 12561112 TI - Ab initio conformational space study of model compounds of O-glycosides of serine diamide. AB - Relative stabilities of rotamers of the N-acetyl-O-(2-acetamido-2-deoxy-alpha-D galactopyranosyl)-L-seryl-N'-methyl amide (1) and eleven analogous molecules containing beta-galactose, alpha- and beta-mannose, alpha- and beta-glucose, and L-threonine were calculated to learn whether they could explain the natural preference for 1 in linkages between the carbohydrate and protein in glycoproteins. The lowest energy rotamers of four O-glycoside models of serine diamide were identified with a Monte Carlo search coupled with molecular mechanics (MM2*). These rotamers were further optimized with an ab initio level of theory (HF/6-31G(d)). Subsequently, B3LYP/6-31 + G(d) single point energies were calculated for the most stable HF structures. The most favorable interactions are present in 1 and its glucose analogue. The monosaccharide for the carbohydrate antenna is anchored to the serine residue with an AcNH...O=C NHMe hydrogen bond in the most stable rotamers. The mannose analogue and the beta anomers are considerably less stable according to the MM2* and especially to the ab inito energy values. The three analogues have HF/6-31 G(d) energies which are 4-6 kcal mol-1 higher; the single point B3LYP/6-31 + G(d)//HF/6-31 G(d) calculations yield preferences of 3-5 kcal mol-1 for 1. The most stable L threonine analogues show a behaviour very similarly to the corresponding serine analogues. The ZPE and thermal correction components of the calculated delta H298 and delta G298 values are relatively small (< 0.4 kcal mol-1). However, the T delta S298 term can be as large as 2.6 kcal mol-1. The entropy terms stabilize the alpha-anomers relative to beta-anomers, and ManNAc relative to GalNAc. The largest stabilization effect is observed for one of the rotamers of the alpha anomer of ManNAc. PMID- 12561114 TI - Core-substituted naphthalene bisimides: new fluorophors with tunable emission wavelength for FRET studies. AB - Highly colored and photoluminescent naphthalene bisimide dyes have been synthesized from 2,6-dichloronaphthalene bisanhydride 1 by means of a stepwise nucleophilic displacement of the two chlorine atoms by alkoxides and/or alkyl amines. The alkoxy-substituted derivatives are yellow dyes with green emission and low photoluminescence quantum yields, whereas the amine-substituted derivatives exhibit a color range from red to blue with strong photoluminescence up to 76%. Structure-property relationships for this class of two-dimensional chromophores were evaluated based on a single-crystal X-ray analysis for dye 5a, the observed solvatochromism, and quantum-chemical calculations. Owing to the simple tuning of the absorption properties over the whole visible range by the respective substituents, the pronounced brilliancy, and the intense photoluminescence, this class of dyes is considered to be highly suited for numerous applications such as fluorescent labeling of biomacromolecules and light harvesting in supramolecular assemblies. As an important step towards such applications efficient FRET (fluorescence resonance energy transfer) has been demonstrated for a covalently tethered bichromophoric compound that contains a red and a blue naphthalene bisimide dye. PMID- 12561115 TI - Highly stable and biocompatible nafion-based capsules with controlled permeability for low-molecular-weight species. AB - Biocompatible hollow capsules have been formed by electrostatic layer-by-layer self-assembly of a perfluorinated ionomer (Nafion) in alternation with ferric ions onto polystyrene latex particles or organic microcrystals, followed by dissolution of the cores by tetrahydrofuran or dimethylformamide. The stepwise growth of multilayers was followed by UV-visible spectroscopy and microelectrophoresis. The formation of hollow capsules was verified by confocal laser scanning microscopy and scanning force microscopy. The hollow Fe3+/Nafion capsules displayed high stability over a wide range of pH values and at high temperature. Fluorescein transport through the Fe3+/Nafion capsule wall was studied by means of photochemical bleaching and recovery (PBR) of the capsule interior. A diffusion model is suggested to calculate the diffusion coefficient for low-molecular-weight species, which was determined to be in the order of 10( 12) cm2s. The permeability can be manipulated by changing the wall thickness of the capsules. PMID- 12561116 TI - The nature of the monomer insertion step in the allylnickel(II)-catalyzed 1,4 polymerization of 1,3-butadiene: sigma-allyl-insertion mechanism versus pi-allyl insertion mechanism. AB - We present a theoretical investigation on the nature of the monomer insertion step in the allylnickel(II)-catalyzed 1,4-polymerization of 1,3-butadiene that employed a gradient-corrected DFT method. We have explored critical elementary steps of the whole polymerization cycle for the trans-1,4 regulating cationic allylnickel(II) [RC3H4NiII(C4H6)L]+ catalyst. These steps are i) cis-butadiene insertion into either the eta 1-sigma-butenyl-NiII bond (sigma-allyl insertion mechanism) or the eta 3-pi-butenyl-NiII bond (pi-allyl insertion mechanism) along with competing pathways for generation of trans-1,4 and cis-1,4 polymer units, and ii) anti-syn isomerization. Based on the analysis of geometric and electronic structures of key species involved and the energetics, we present a detailed insight into the different nature of the monomer insertion step according to the two mechanistic alternatives. An understanding of why the pi-allyl insertion mechanism is favored over the sigma-allyl insertion mechanism is provided. eta 1 sigma-butenyl-NiII Species are predicted to be sparsely populated and also distinctly less reactive than eta 3-pi-butenyl-NiII species. Although they are commonly believed to be reactive intermediates, eta 1-sigma-butenyl-NiII species are, therefore, not likely to be involved along viable pathways for cis-butadiene insertion into the butenyl-NiII bond. The present investigation corroborates our previous conclusion that the pi-allyl insertion mechanism represents the preferred mechanism for the monomer insertion step in the allylnickel(II) catalyzed 1,4-polymerization of 1,3-butadiene. On the other hand, the suggested alternative sigma-allyl insertion mechanism has to be considered to be not operative, for both thermodynamic and kinetic reasons. Furthermore, the sigma allyl insertion mechanism would neither provide a rationalization of cis-trans selectivity nor of chemoselectivity in the allylnickel(II)-catalyzed 1,4 polymerization of 1,3-butadiene. PMID- 12561117 TI - Solid-phase development of chiral phosphoramidite ligands for enantioselective conjugate addition reactions. AB - The development of a method for the optimization of chiral ligands for the steric steering of enantioselective Cu-catalyzed conjugate additions of Znalkyls to enones is described. The method is based on combinatorial principles and solid phase techniques. It includes the combinatorial synthesis of chiral bispidine derived ligands embodying a phosphoramidite group on the solid phase and their investigation in immobilized form in the conjugate addition of ZnEt2 to cyclohexenone as test reaction. The best identified ligands were also synthesized separately and investigated in its soluble form. The results obtained for the polymer-bound ligands correctly mirrored the performance of the soluble ligands. The library embodied members giving ee values varying between 3 and 67%. The "positional scanning" approach proved to be invalid for the study of the ligand system, indicating that this approach in general should be applied with care. Taken together, the method allowed for rapid and efficient optimization of the ligands and led to the development of the first enantioselective, Cu-catalyzed conjugate addition reaction with a polymer-bound ligand. PMID- 12561118 TI - Molecular design of a pyrrole-imidazole hairpin polyamides for effective DNA alkylation. AB - New hairpin polyamide-CPI (CPI = cyclopropylpyrroloindole) conjugates, compounds 12-14, were synthesized and their DNA-alkylating activities compared with the previously prepared hairpin polyamide, compound 1, by high-resolution denaturing gel electrophoresis with 450 base pair (bp) DNA fragments and by HPLC product analysis of the synthetic decanucleotide. In accord with our previous results, alkylation by compound 1 occurred predominantly at the G moiety of the sequence 5'-AGTCAG-3' (site 3). However, compound 12, in which the structure of the alkylating moiety of compound 1 is replaced with segment A of duocarmycin A DU-86 (CPI), did not show any DNA alkylating activity. In clear contrast, the hairpin CPI conjugate 13, which differs from compound 1 in that it lacks one Py unit and possesses a vinyl linker, alkylated the A of 5'-AGTCAG-3' (site 3) efficiently at nanomolar concentrations. Alkylation by compound 14, which has a vinyl linker, occurred at the A of 5'-AGTCCA-3' (site 6) and at several minor alkylation sites, including mismatch alkylation at A of 5'-TCACAA-3' (site 2). The significantly different reactivity of the alkylating hairpin polyamides 1, 12, 13, and 14 was further confirmed by HPLC product analysis by using a synthetic decanucleotide. The results suggest that hairpin polyamide--CPI conjugate 13 alkylates effectively according to Dervan's pairing rule, and with a new mode of recognition in which the Im-vinyl linker (L) pair targets G-C base pairs. These results demonstrate that incorporation of the vinyl-linker pairing with Im dramatically improves the reactivity of hairpin polyamide--CPI conjugates. PMID- 12561119 TI - Monodisperse CdSe nanorods at low temperatures. AB - A new synthetic method is presented that allows the preparation of highly monodisperse CdSe nanorods (so called quantum rods) at relatively low temperatures (160 degrees C). This method is characterized by a high aspect ratio of the particles and affords good reproducibility. The morphology of the resulting nanorods was examined by means of transmission electron microscopy (TEM) and the electro-optic properties by means of fluorescence spectroscopy. The conditions of the reaction of nanoparticle growth were examined by varying the concentration of the organometallic precursors, the growth temperature, and the growth time. The experimental findings correspond well with previously published semiempirical pseudo-potential calculations. PMID- 12561120 TI - Stable oligonucleotide-directed triplex formation at target sites with CG interruptions: strong sequence-specific recognition by 2',4'-bridged nucleic-acid containing 2-pyridones under physiological conditions. AB - A sequence of double-stranded DNA (dsDNA) which can be recognized by a triplex forming oligonucleotide (TFO) is limited to a homopurine-homopyrimidine sequence. To develop novel nucleoside analogues which recognize CG interruption in homopurine-homopyrimidine dsDNA, we synthesized a novel 2'-O,4'-C methyleneribonucleic acid (2'-O,4'-C-methylene bridged nucleic acid; 2',4'-BNA) that bears the unnatural nucleobases, 2-pyridone (PB) or its 5-methyl congener (mPB); these analogues were introduced into pyrimidine TFOs using a DNA synthesizer. A TFO with a 2'-deoxy-beta-D-ribofuranosyl-2-pyridone (P) or 2',4' BNA abasic monomer (HB) was also synthesized. The triplex-forming ability of various synthesized 15-mer TFOs and the corresponding homopurine-homopyrimidine dsDNA, which contained a single pyrimidine-purine (PyPu) interruption, was examined in UV melting experiments. It was found that PB and mPB in the TFOs successfully recognized CG interruption under physiological conditions (7 mM sodium phosphate, 140 mM KCl, 5 mM spermine, pH 7.0). Furthermore, triplex formation between the dsDNA target which contained three CG interruptions and the TFO with three PB units was also confirmed. Additional four-point 2',4'-BNA modifications of the TFO containing three PB units significantly enhanced its triplex-forming ability towards the dsDNA and had a Tm value of 43 degrees C under physiological conditions. These results indicate that a critical inherent problem of TFOs, namely, the sequence limitation of the dsDNA target, may be overcome to a large extent and this should promote antigene applications of TFOs in vitro and in vivo. PMID- 12561121 TI - Design and structural extension of a supramolecular inclusion-compound host made by the formation of dimers of isonicotinic acid and thiocyanato coordinating bridges. AB - A new host design for an inclusion compound with a preference for large planar aromatic guest molecules has been proposed. Our host design includes a rectangular cavity made using a long and a short building block based on the concept of supramolecular chemistry. The long building block facilitates the inclusion of large guests, and the short building block prevents the formation of an interpenetrated structure, which is often observed in frameworks with large void spaces. The long building block is made when dimers of 4-pyridinecarboxylic acid (isoH) form through hydrogen bonding between the two carboxylic acid moieties. This isoH dimer can link two transition metal centers using the N atoms at both ends to act as a long building block. For the short building block, the thiocyanato ion was used. This makes a bent bridge between two metal centers to form a 1D double-chain [M(SCN)2]infinity complex. From the self-assembly of isoH, SCN- and Ni2+, a 2D network of [Ni(SCN)2(isoH)2]infinity, in which the 1D [Ni(SCN)2]infinity complexes are linked by the isoH dimers, is built up. The rectangular cavity is formed as a mesh within the 2D network. The crystal of our inclusion compound has a layered structure of 2D networks, and a 1D channel-like cavity penetrating the layered 2D networks is formed where guests may be included. Moreover, our host design has the advantage of easy extension of the host structure. Replacement of isoH with another component and use of three components is possible for making the long building block. In the latter case, a linear spacer having two carboxy groups is inserted into the isoH dimer to form a long building block with a trimer structure. Based on our host design, a series of new inclusion compounds were synthesized. The crystal structures of three compounds were determined by single crystal X-ray diffraction. These were a biphenyl inclusion compound [Ni(SCN)2(isoH)2].1/2C12H10 (the basic case), a 9,10 dichloroanthracene inclusion compound [Ni(SCN)2(acrylH)2].1/2C14H8Cl2, where isoH is replaced with 3-(4-pyridinyl)-2-propenoic acid (acrylH), and a perylene inclusion compound [Ni(SCN)2(isoH)2(fumaricH2)].1/2C20H12, whose long building block is a trimer inserted with fumaric acid (fumaricH2) as a linear spacer. PMID- 12561122 TI - Double-stranded helices and molecular zippers assembled from single-stranded coordination polymers directed by supramolecular interactions. AB - Using three nonlinear dicarboxylates, isophthalate (ipa), 4,4'-oxybis(benzoate) (oba), and ethylenedi(4-oxybenzoate) (eoba), we have prepared five neutral infinite copper(II) dicarboxylate coordination polymers containing lateral aromatic chelate ligands, namely [Cu(ipa)(2,2'-bpy)]n.2nH2O (1), [Cu2(ipa)2(phen)2H2O]n (2), [Cu(oba)(phen)]n (3), [Cu(oba)(2,2'-bpy)]n (4), and [Cu(eoba)(phen)]n (5; 2,2'-bpy = 2,2'-bipyridine, phen = 1,10-phenanthroline) by hydrothermal synthesis. X-ray single-crystal structural analyses of these complexes reveal that the nonlinear flexible or V-shaped dicarboxylates can induce the helicity or flexuousity of the polymeric chains and aromatic chelate ligands are important in providing potential supramolecular recognition sites for pi-pi aromatic stacking interactions. An appropriate combination of the bridging dicarboxylate and aromatic chelate can induce a pair of single-stranded helical or flexuous chains to generate a double-stranded helix or molecular zipper through supramolecular interactions, respectively. PMID- 12561123 TI - After years of waiting, health care finally has a Baldrige winner! AB - SSM Health Care was not an "overnight success;" quality journey has taken many years. Health care faced unique challenges in competing with industry. Baldrige standards helped SSM crystallize mission, goals. PMID- 12561124 TI - SSM health care shares its best practices. PMID- 12561125 TI - Quality focus shining on corporate ethics. AB - Compliance just scratches the surface of a comprehensive ethics policy. Being true to your mission is a critical component of governance responsibilities. Quality managers play an important role in feedback, implementation. PMID- 12561126 TI - Successful practices benchmarked at CHA. AB - The goal was empowerment of members to move on to more successful practices. Employee involvement in decision making was a key area of focus. Several facilities model identified best practices successfully. PMID- 12561127 TI - U.S. end-of-life care gets a (barely) passing grade. AB - The vast majority of Americans would prefer to die at home. A minority of hospitals offer hospice or palliative care services. Report card enables state-to state comparisons, benchmarking. PMID- 12561128 TI - Med school teaches bioterror response. AB - Subtle, natural course changes are seen as most effective. Students are trained to respond to "all events," not just specific attacks. Efforts are under way to enable benchmarking by other institutions. PMID- 12561129 TI - An analysis of response, direction, and place learning in an open field and T maze. AB - Rats were trained to locate food in a response, direction, or place problem on an open field located at 2 positions. In Experiment 1, both the response and direction groups solved the problem. The place group failed to solve the task in approximately 300 trials. Experiment 2 demonstrated that rats need distinguishable start points to solve a place problem when neither a response nor a direction solution is available. Findings from Experiment 3 suggest that a combination of path traveled and distinct cues help to differentiate start points. Experiment 4 replicated the findings using a T maze. These results suggest "place" solutions are difficult for rats. The data are discussed with respect to conditional learning and modern spatial mapping theory. PMID- 12561130 TI - Interacting Cache memories: evidence for flexible memory use by Western Scrub Jays (Aphelocoma californica). AB - When Western Scrub-Jays (Aphelocoma californica) cached and recovered perishable crickets, N. S. Clayton, K. S. Yu, and A. Dickinson (2001) reported that the jays rapidly learned to search for fresh crickets after a 1-day retention interval (RI) between caching and recovery but to avoid searching for perished crickets after a 4-day RI. In the present experiments, the jays generalized their search preference for crickets to intermediate RIs and used novel information about the rate of decay of crickets presented during the RI to reverse these search preferences at recovery. The authors interpret this reversal as evidence that the birds can integrate information about the caching episode with new information presented during the RI. PMID- 12561131 TI - Trial spacing is a determinant of cue interaction. AB - Four conditioned lick suppression experiments with rats examined the effect of trial spacing on cue interaction. Experiments 1 and 2 found overshadowing to be eliminated with massed compound stimulus-outcome pairings and the usual trial spacing effect to be reversed with compound acquisition trials. Experiment 3 found that whether acquisition compound-outcome pairings were massed or spaced determined the effect of posttraining extinction treatment. Extinction of the overshadowing cue reduced responding following massed training and increased responding following spaced training. Extinction of the context decreased responding following massed training. Experiment 4 found the conditioning and devaluation results to be associative and stimulus specific. These results are in accord with the extended comparator hypothesis (J. C. Denniston, H. I. Savastano, & R. R. Miller, 2001). PMID- 12561132 TI - Perceptual learning in flavor aversion: evidence for learned changes in stimulus effectiveness. AB - Rats were exposed to the compound flavors AX and BX, presented in alternation, and to CX on a separate block of trials. Generalization to BX after aversion conditioning with AX was less than to CX. An equivalent effect was found when the nature of the common element was changed after preexposure but not when the common element was omitted during preexposure, during conditioning and test, or both. Rats conditioned with X alone again showed less aversion to BX than to CX; similarly, rats conditioned with a novel flavor (Y) showed less aversion to BY than to CY. These effects support the proposal that intermixed preexposure to AX and BX enhances the perceptual effectiveness of their unique features, A and B. PMID- 12561133 TI - Complex dynamic processes in sign tracking with an omission contingency (negative automaintenance). AB - Hungry pigeons received food periodically, signaled by the onset of a keylight. Key pecks aborted the feeding. Subjects responded for thousands of trials, despite the contingent nonreinforcement, with varying probability as the intertrial interval was varied. Hazard functions showed the dominant tendency to be perseveration in responding and not responding. Once perseveration was accounted for, a linear operator model of associative conditioning further improved predictions. Response rates during trials were correlated with the prior probabilities of a response. Rescaled range analyses showed that the behavioral trajectories were a kind of fractional Brownian motion. PMID- 12561134 TI - Importance of trials versus accumulating time across trials in partially reinforced appetitive conditioning. AB - Four experiments with rats examined partial reinforcement in appetitive conditioning. In Experiment 1, adding nonreinforced trials to a continuous reinforcement schedule slowed acquisition, whereas deleting reinforcers did not. Trial massing suppressed performance and learning. In Experiment 2, conditioning with a short conditioned stimulus (CS) was rapid, and partial reinforcement with a short CS was as effective as continuous reinforcement with equal accumulated time in the CS. In Experiment 3, conditioning was nevertheless influenced by the probability of reinforcement. In Experiments 3 and 4, conditioning was especially disrupted when nonreinforced trials preceded reinforced trials closely in time. The results underscore the importance of temporal variables in conditioning but are more consistent with trial-based accounts than time-accumulation accounts of conditioning. PMID- 12561135 TI - Differential effects of adding and removing components of a context on the generalization of conditional freezing. AB - Rats were conditioned across 2 consecutive days where a single unsignaled footshock was presented in the presence of specific contextual cues. Rats were tested with contexts that had additional stimulus components either added or subtracted. Using freezing as a measure of conditioning, removal but not addition of a cue from the training context produced significant generalization decrement. The results are discussed in relation to the R. A. Rescorla and A. R. Wagner (1972), J. M. Pearce (1994), and A. R. Wagner and S. E. Brandon (2001) accounts of generalization. Although the present data are most consistent with elemental models such as Rescorla and Wagner, a slight modification of the Wagner-Brandon replaced-elements model that can account for differences in the pattern of generalization obtained with contexts and discrete conditional stimuli is proposed. PMID- 12561136 TI - Pigeons learn stimulus identity and stimulus relations when both serve as redundant, relevant cues during same-different discrimination training. AB - The authors taught pigeons to discriminate displays of 16 identical items from displays of 16 nonidentical items. Unlike most same-different discrimination studies--where only stimulus relations could serve a discriminative function- both the identity of the items and the relations among the items were discriminative features of the displays. The pigeons learned about both stimulus identity and stimulus relations when these 2 sources of information served as redundant, relevant cues. In tests of associative competition, identity cues exerted greater stimulus control than relational cues. These results suggest that the pigeon can respond to both specific stimuli and general relations in the environment. PMID- 12561137 TI - Euthanasia and surgery. PMID- 12561138 TI - Euthanasia. PMID- 12561139 TI - Opinion of the National Council on Euthanasia. Meeting of the National Council of 15th January 2000. PMID- 12561140 TI - Bulletin of the National Council, 9th December 2001. Euthanasia (17/11/2001). PMID- 12561141 TI - [The law relative to euthanasia: conciliate the ethics]. PMID- 12561142 TI - [Euthanasia debate]. PMID- 12561143 TI - The Belgian law on euthanasia. The historical and ethical background. AB - A survey is provided of the genesis of the euthanasia laws in Belgium and the Netherlands, with the intent to provide a clear idea of the concepts involved and of the rationale for their introduction. The role of the Belgian Consultative Committee on Bioethics in the development of the ethical discussion in Belgium is explained as well as the essence of the profound ethical divide between the 'coalition' and the Christian Democrats. After a summary of the basic points of the law, a comparison is made between the Belgian and the Dutch laws and their ethical foundations. PMID- 12561145 TI - [The role of palliative care in the health care system]. PMID- 12561144 TI - [The problem of end of life intensive care and the new law on euthanasia]. PMID- 12561146 TI - The protection of the patient's private life: the computer challenge. Second part. AB - Today, medical practice is invaded by a growing number of technologies of all kinds, among which computer techniques have an important place. Although they have significant advantages, for instance in terms of medical record management, they give rise to several problems, particularly concerning the confidentiality of the patient's data with regards to third party. A great number of specific provisions, complementary to the general texts protecting private life (examined in the first part of this two parts article), endeavour to solve these problems. It is true that these provisions are recent, have various origins and often appear as rules difficult to understand. Yet, they are partially inspired by a common logic. Relying on these common features, the authors make two suggestions for the future, in order to avoid that the growing computerisation of medical practice eventually destabilises the health care relationship: a) Any dictatorship of confidentiality must be rejected b) Stimulating a sense of professionalism is most likely the way to avoid an anarchic and unrealistic development of rules aimed at regulating the health care relationship. PMID- 12561147 TI - Carotid endarterectomy: looking for the gold standard. PMID- 12561148 TI - Carotid artery stenting: which stent for which lesion? AB - Although some early reports describe angiographic as well as clinical success for balloon angioplasty alone in the treatment of carotid occlusive disease, most interventionists prefer stent-assisted balloon angioplasty because of the purported advantages, such as avoiding plaque dislodgement, intimal dissection, elastic vessel recoil and late restenosis. Mainly because of the different characteristics of each carotid artery segment, different types of stents are preferred. A carotid artery lesion located in the intrathoracic brachiocephalic trunc or common carotid artery would need a different stent to a lesion at the carotid bifurcation or a lesion of an intracerebral branch of the internal carotid artery. PMID- 12561149 TI - Causes of early post carotid endartectomy stroke in a recent series: the increasing importance of hyperperfusion syndrome. AB - OBJECTIVE: Early stroke occurs in 0.9% to 7% of patients undergoing carotid endarterectomy (CEA). These have been thought to be mostly due to embolization. However, in our recent clinical experience, we noted hyperperfusion syndrome to be a significant cause of postoperative strokes. Therefore, we reviewed our experience and investigated the distribution of causes of early postoperative strokes. MATERIALS AND METHODS: A retrospective chart review of 444 consecutive patients who underwent CEA at our institution between June 1997 and October 1999 (500 operations) was performed to evaluate the incidence and etiology of early postoperative strokes. Indications for operation included history of stroke correlating with the side of ICA stenosis (50 patients or 10%), symptoms of transient ischemic attacks (84 patients or 16.8%), amaurosis fugax (18 patients or 3.6%), or asymptomatic stenosis (348 patients or 69.6%). All patients were evaluated with duplex scan preoperatively and postoperatively. Diagnosis of early postoperative strokes within one month after CEA was made based on clinical examination. Postoperative CT scan of the brain was available in 100% of patients with suspected diagnosis of CVA. RESULTS: Five patients (3 male and 2 female) were diagnosed with strokes postoperatively (1%). These five were symptomatic patients with ICA stenosis > or = 80% and moderate contralateral ICA stenosis. In two of those patients (40%), the reason for the stroke was considered embolization to the cerebral arteries; one patient suffered a shunt injury as a cause of stroke; two patients (40%) were diagnosed with hyperperfusion syndrome. In both patients diagnosis was made clinically, and in only one of those patients the clinical picture correlated with CT scan. Two patients (0.4%) had asymptomatic ICA occlusion at 2 weeks and 18 days postoperatively. CONCLUSION: Embolization to the cerebral arteries remains the leading cause of early postoperative strokes. However, hyperperfusion syndrome also accounts for a significant portion of these postoperative strokes. The percentage of patients with this syndrome might be even higher, once clinical picture is clearly defined. These data warrant further investigation of hyperperfusion syndrome. PMID- 12561150 TI - Long-term results after video-assisted thoracic surgery for spontaneous pneumothorax. AB - OBJECTIVE: Analysis of the long-term results of video-assisted thoracic surgery (VATS) for spontaneous pneumothorax in patients treated in two surgical centres from May 1994 until December 2000. METHODS: A cohort of 86 patients was studied retrospectively. For final analysis, 74 patients undergoing 76 VATS procedures were included. RESULTS: Sixty-three procedures for primary spontaneous pneumothorax (PSP) and 13 procedures for secondary spontaneous pneumothorax (SSP) were performed. In 78.9% (n = 60) blebs or bullae were resected with an endostapler device. In 2.6% (n = 2) an apical fibrotic zone was resected. In 71.1% (n = 54) a subtotal pleurectomy was performed combined with an abrasion in 41 cases. In 21.1% (n = 16) a total pleurectomy and in 7.8% (n = 6) an abrasion was performed. Operative mortality was 1.3% (n = 1) and total operative morbidity 25.4% in the PSP group and 76.9% in the SSP group. Mean follow-up was 36 months. The global recurrence rate was 5.3% (n = 4), being 4.8% (n = 3) in the PSP group and 7.7% (n = 1) in the SSP group. Recurrences occurred at 1 (n = 2), 2 (n = 1) and 16 (n = 1) months after the initial operation. The incidence of postoperative neuralgia was 17.1% (n = 13). One patient needed analgesics for the neuralgia. CONCLUSION: VATS treatment of spontaneous pneumothorax proves to be effective. In cases of SSP, VATS treatment is feasible but a higher morbidity rate should be anticipated. PMID- 12561151 TI - Emergency aorto-iliac aneurysm surgery with low mortality and morbidity. AB - A retrospective review of urgently operated aortic or iliac aneurysms over a 13 1/2 year period identified 51 patients (50 male, one female). In our consecutive series, 45 patients underwent an emergency operation for an abdominal aortic aneurysm (AAA) and six patients for an iliac aneurysm (IA). Mean age was 69 years. All patients had prominent symptoms: acute low abdominal pain or low back pain in 20 patients, shock in six patients, shock and pain in 25 patients. Free rupture was found in 28 cases, retroperitoneal rupture in 14 cases, fissurisation in seven and arterio-venous fistulisation in two cases. All reconstructions were done by the same vascular surgeon using Dacron prostheses. Intra-operative mortality rate was 3.9% (n = 2), 30-day mortality was 21.6% (n = 11) and cumulative hospital mortality was 23.5% (n = 12). The morbidity was 59%. PMID- 12561152 TI - Repair of atrial septal defects via limited right anterolateral thoracotomy. AB - INTRODUCTION: Right thoracotomy is a well known alternative for median sternotomy to gain access to the right atrium. The Port-Access technique is a surgical option to reduce the skin incision to 5 cm and have a considerable gain in cosmesis and post-operative rehabilitation. PATIENTS AND METHODS: From February 1997 to May 2000, 50 patients (19M/31F) had Port-Access atrial septal defect repair (ASD) with the Heartport Endo-CPB and Endo-aortic clamp system. Mean age was 46 years (range 10.5-74 years). Forty-nine patients had a type II ASD. Most of the patients were asymptomatic (48 were NYHA class I-II). Five patients underwent combined procedures: two mitral valve repairs, one mitral valve replacement, one plasty of the superior vena cava and one mini Cox-Maze. Direct closure was obtained in 34 patients (68%). Mean aortic cross clamp time was 56 minutes (range 24-134 min) and mean perfusion time was 102 minutes (range 32-196 min). RESULTS: There were no conversions to sternotomy. The procedure was complicated in six patients: revision for bleeding (n = 1), stenting of the iliac vein (n = 1), enlargement plasty of the femoral artery (n = 1), transient renal failure (n = 1), sick sinus syndrome requiring pacemaker implantation (n = 1) and one lymphocoele. No thromboembolic or peripheral ischaemic complications were noted. Per- and postoperative echocardiography showed no leakage in any patients. There was no hospital mortality. Mean ICU and hospital stay were 1.14 days (range 1-3 days) and 6.41 days (range 4-10 days) respectively. CONCLUSION: The Port Access approach of ASD closure constitutes a valid alternative to sternotomy with the same standards of results and quality. PMID- 12561153 TI - Dermatofibrosarcoma protuberans: how wide is wide in surgical excision? AB - Dermatofibrosarcoma protuberans is a rare mesenchimal tumour mainly characterized by its local aggressiveness and high reported recurrence rates. Classically, wide excisional surgery with at least 3 cm of free tissue margins is recommended as treatment. We present a series of 18 patients with variable gross free tissue margins but confirmed histological free margins in all cases whatever the surgical excision really was. Only one patient presented a local recurrence (5.5%) of a very large (14 cm) sarcomatous variant of dermatofibrosarcoma protuberans. Mean follow-up in this series was 52.1 months. Those results emphasize that histological rather than theoretical "golden rules" are mandatory to locally control this aggressive neoplasm. PMID- 12561154 TI - False aneurysm of the hepatic artery after laparoscopic cholecystectomy. AB - PURPOSE: A rare complication of laparoscopic cholecystectomy is defined: iatrogenic injuries to hepatic artery system which may evolve to pseudoaneurysms in the late postoperative period. This rare phenomenon may be overlooked and pose a challenge to surgeons. MATERIAL AND METHODS: We will describe three cases with iatrogenic pseudoaneurysms after laparoscopic cholecystectomy. The onset of symptoms and the course of the disease was not uniform. Diagnosis was made after a considerable delay. In the first case, a small, uncomplicated extrahepatic pseudoaneurysm was successfully treated with coil embolization. The second patient who had an intrahepatic pseudoaneurysm with multiple injuries to the common bile duct and portal vein, did not survive despite surgical and endovascular interventions. In the latter, surgical treatment for a large pseudoaneurysm that had ruptured into the liver parenchyma was successfully conducted. Review of the literature reveals fifty-four more cholecystectomy related pseudoaneurysms. The site of injury was the right hepatic artery in 61% of the cases and the presenting symptom was upper gastrointestinal bleeding (haemobilia) in two-third of the patients. Embolization was performed in 82% of the cases, and surgery was undertaken in the remaining 18%. CONCLUSION: Pseudoaneurysm is an uncommon complication of laparoscopic cholecystectomy. Prompt attention is necessary since the lesion has a high risk of rupture. Embolization is the first line of treatment and surgery is reserved for more complex injuries and cases with life-threatening rupture of the aneurysm. PMID- 12561155 TI - Giant ancient schwannoma of the posterior mediastinum cytologically misdiagnosed as a malignant tumour. A case report. AB - We report a case of a 45-year old woman who was found to have a giant mediastinal tumour with radiological degenerative changes. She underwent thoracotomy to remove the mass, which was eventually diagnosed histologically as an ancient schwannoma, whereas cytological interpretation of the accompanying pleural fluid was malignant. Ancient schwannoma is a rare variant of schwannoma, histologically showing atypical features that may result in erroneous diagnosis of a malignant tumour. Clinical and radiological findings are important aids for further consideration of surgical removal of these potentially resectable tumours. PMID- 12561156 TI - Popliteal artery pseudo-aneurysm and hereditary multiple exostoses. AB - We describe a rare case of a 21-year-old man presenting with hereditary multiple exostosis and a pseudoaneurysm of the popliteal artery caused by femoral osteochondroma. Principles of management and surgical technique are discussed. PMID- 12561157 TI - Nocardia asteroides cerebral abscess in a renal transplant recipient: short report. AB - Opportunist central nervous system infections occur in about 5% to 10% of all renal transplant recipients, but reports of brain abscesses are very rare (1). Nocardia asteroides cerebral abscesses are scarce intracranial lesions. They account for only 2% of brain abscesses (2). Published data about these lesions have taken the form of short reports, small cases series and reviews. A universally accepted and effective treatment approach has not yet been established. We present a renal transplant patient with a cerebral abscess caused by Nocardia asteroides. PMID- 12561158 TI - Tuberculosis in Canada 2000: executive summary. PMID- 12561159 TI - Tuberculosis among the foreign-born in Canada. AB - Increasingly, reducing the burden of TB disease in Canada will depend on controlling the disease in foreign-born people. The foreign-born have a greatly elevated risk of TB disease within the first 2 years after arrival, but the risk of developing TB is still present for many years afterwards. Drug resistant TB is seen predominantly in this population but has not yet become a problem in Canada. PMID- 12561160 TI - [Regulation effect of forest vegetation on watershed runoff in the Loess Plateau]. AB - The runoff and rainfall data of typical forested and non-forested watersheds in Ziwuling region, which is located in the hinterland of the Loess Plateau, were used to analyze their annual runoff variation comparatively. The result showed that the annual distribution of runoff in forested watersheds was uniform, compared with that in non-forested watersheds. The total runoff in flood season decreased by 8.88 mm and 7.1 mm. Regression analyses between runoff in dried-up season and rainfall in flood season or in dried-up season showed that the rainfall in flood season in forested watersheds could be changed into underground runoff by forest vegetation, and increased the runoff in dried-up season. The annual variation of runoff also indicated that the total effect of forest vegetation on dried-up runoff complement from October to December was 1.69 mm and 0.5 mm, and that from January to May was not significant. PMID- 12561161 TI - [Community characteristics of Hippophae rhamnoides forest and water and nutrient condition of the woodland in Loess hilly region]. AB - To improve the productivity of Hippophae rhamnoides forest and to manage the forest reasonably, the community characteristics of Hippophae rhamnoides and the water and nutrient condition of the woodland were analyzed. The results showed that Hippophae rhamnoides grew fast and its adaptability was strong. 4-5 years old Hippophae rhamnoides could form shrubs-grass community. The biomass 1-8 years old Hippophae rhamnoides increased rapidly, but that of 8-11 years old increased more slowly. Hippophae rhamnoides older than 11 years could keep reasonable community structure and higher primary productivity by natural thinning. The utilization of soil moisture by Hippophae rhamnoides could be classified into four layers as faint utilization layer of root system (0-20 cm), utilization of root system (20-300 cm), adjustment layer supplied by soil moisture (300-400 cm), and faint adjustment layer (400-500 cm). Due to the soil improvement by Hippophae rhamnoides and the water retaining by litter, the soil moisture of 1-1.5 m layer could be easy to restore. Hippophae rhamnoides could keep the balance of nutrition in the woodland through the fixation of nitrogen by root nodule and the decomposition of litter. In the 6-9 years old Hippophae rhamnoides plantation, the total nitrogen content of soil might be improved from 0.05-0.1% to 0.2%. PMID- 12561162 TI - [Comparative analysis on species diversity of hillclosed afforested plant community in Beijing Jiulong Mountain]. AB - The plant communities in Beijing Jiulong Mountain were surveyed with the Quadrate. The species diversity characteristics of the tree layer and the shrub layer and herb layer of these plant communities were measured by applying Shannon Wiener diversity index(D), evenness index(J) and ecological dominance(C), and the comparison of the characteristics was conducted on shade slope and sun-faced slope. The results showed that these indices could effectively reveal the characteristics of species composition for plant communities in warm temperate zone. On shade slope or sun-faced slope, the D value of shrub layer in Jiulong shrub communities (SC) was higher than that of tree layer in arbor forest (AF), but lower than that of shrub layer in AF; the D value of herb layer in SC was lower than that of herb layer in AF; and the D value of herb layer was higher than that of shrub layer in AF, which was higher than that of tree layer in AF. C value presented quite the opposite with D value, and J presented the different feature in the comparison, respectively on shade slope and sun-faced slope. In addition, the succession series and the restoration measures on accelerating the progressive succession of the hillclosed afforested vegetation communities were tentatively discussed. PMID- 12561163 TI - [Storage and decomposition of fallen wood in dark coniferous forest on the north slope of Changbai Mountain]. AB - The storage, biomass, and decomposition of fallen woods in dark coniferous forest on the north slope of Changbai Mountain at elevations of 1260 m and 1620 m were studied. The results showed that the volume of fallen woods was 180.87 m3.hm-2, occupying 21.83% of the stand volume. The volume and biomass of fallen woods at elevation of 1260 m were higher than those at 1620 m. The volume of fallen woods was 52.57 m3.hm-2 at 1260 m, and was 193.85 m3.hm-2 at 1620 m, and their biomass was 6.21 t.hm-2 at 1260 m, and was 53.33 t.hm-2 at 1620 m. Monomial exponential attenuation model could be used to simulate the decomposition process of fallen woods. The results indicated that the decomposition constants of fallen woods were different among different tree species and different elevations. The decomposition constant of fallen woods of Abies was higher than that of Picea, and was higher at 1260 m than at 1620 m. The results also showed that the decomposition of Picea lasted longer. PMID- 12561164 TI - [Regulation function of calcium on photosynthesis of Dimocarpus longana Lour. cv. wulongling under simulated acid rain stress]. AB - Studies on the regulation function of calcium on photosynthesis of Dimocarpus longana under simulated acid rain stress showed that the photoreduction activity of chloroplasts was activated when the concentration of calcium ion in reaction medium ranged from 0 to 5 mmol.L-1, and peaked at the 3.5 mmol.L-1, which was 41.90% higher than that of control. Conversely, the activity of chloroplasts reduced 26.06% in the reaction medium with a concentration of 2 mmol.L-1 EGTA, as compared with the control. Both Mn2+ and Mg2+ could inhibit photoreduction activity. The photophosphorylation activity increased when the concentration of calcium ion in reaction medium ranged from 0 to 6 mmol.L-1, and peaked at the 4.5 mmol.L-1, while superoxidase dismutase (SOD) activity rose from 0 to 6 mmol.L-1 and peaked at 6 mmol.L-1. Calcium ion with the concentration of both 10 mmol.L-1 and 15 mmol.L-1 could increase the content of chlorophyll(Chl), stabilize the membrane structure of leaf discs, and reduce the membrane permeability under simulated acid rain with pH value of 3.0. The effect in 15 mmol.L-1 were better than in 10 mmol.L-1. However, the injury of acid rain to leaves was strengthened when the concentration of calcium was higher than 20 mmol.L-1. Net photosynthesis rate (Pn) rose when leaves sprayed with 15 mmol.L-1 Ca(NO3)2 before treatment of acid rain stress of pH 2.5. All of the results represented the excellent protection function of calcium on D. longana leaves under simulated acid rain. PMID- 12561165 TI - [Biological characteristics of different forest soils in Nanjing-Zhenjiang mountan area]. AB - Studies on the biological characteristics, including soil microbe, soil enzyme activity, soil nutritient content, and litter decomposition of different forest soils in Nanjing-Zhenjiang mountain area showed that the amounts of microbes and the activities of six enzymes in forest soils changed regularly in different forests during different months. The contents of nutritional elements varied regularly with forest growth bio-cycles, and were inerrelated prominently with the amounts of soil microbes and the activities of soil enzymes. There existed temporal-spatial differences in the decomposition rate of litters and the reverted velocity of nutrients among different forests. The comparisons of various biological characteristics among secondary Quercus variabilis forest, Phllostachys pubescens forest, and Cunninghamia lanceolata forest indicated that secondary Quercus variabilis forest had the most abundant nutrients in soil, and possessed the strongest ability of self-fertilization. Therefore, to construct coniferous forests with broadleaf trees in this area could avoid or abate the decline of soil fertility. PMID- 12561166 TI - [Mechanism and universality of Yoda's -3/2 law of self-thinning in plant population]. AB - According to plant modular theory, the Yoda's-3/2 law of self-thinning was theoretically rederived in this paper. With the parameters in the expression of Yoda's-3/2 law being given biological meanings, this article offered universal and rational explanations for relevant data in literatures that could not be explained before. Based on the authors' hypothesis, the experiments on the self thinning process of spring wheat (Triticum aestivum L.) were conducted. Individual morphological and population quantitative indexes were examined at regular intervals. The results showed that the self-thinning process of spring wheat conformed to Yoda's-3/2 law of self-thinning. At the same time, the universality of Yoda's-3/2 law of self-thinning was explained theoretically by analyzing individual morphological data. PMID- 12561167 TI - [A mathematical model of water stress and light condition effects on cotton dry matter and yield formation]. AB - A mathematical model was developed to analyze the effects of water stress and light condition on crop dry matter accumulation and yield formation based on canopy carbon net assimilation rate. The function leaf water potential (psi l) indicating the water status of canopy was incorporated into this model, according to the assumption that the canopy resistance (Rc) was increased under the conditions of water stress and low light density. Psi l was estimated by a simplified regression equation, in which, the independent variables were relative soil moisture (Aw), ambient temperature (Ta), and vapor pressure deficit (VPD). The aerodynamic resistance (Ra) in the model was defined as a function of wind speed (u), and the yield was calculated by a linear increase in harvest index (hi) with time. The modeled data agreed well with the data observed from pot experiment. Sensitivity analysis and simulation results suggested that the model could be useful in identifying environment factors, especially soil water content and light density effects on crop growth and yield formation. PMID- 12561168 TI - [Difference of several major nutrients accumulation in vegetable and cereal crop soils]. AB - Investigation and determination of several major nutrients in different types of vegetable and cereal crop soils were carried out in the Guanzhong Plain in Shaanxi Province. The results showed that organic matter, nitrogen, phosphorus and potassium accumulated more in the 0-200 cm profile of vegetable soils than in that of cereal crop soils. However, the accumulation degrees were different with different nutrient forms. The total amount of nitrate-N in the soil profile of plastic greenhouse and usual vegetable field was 1520.9 kg.hm-2 and 1358.8 kg.hm 2, being 5.2 and 4.5 times higher than that in the cereal crop fields respectively. The total amount of available-P was respectively 978.1 kg.hm-2 and 503.3 kg.hm-2 in the two vegetable soils, and 136.2 kg.hm-2 in cereal crop soils, and the former two were 6.2 and 2.7 times higher than the latter. For other nutrients, organic matter in cereal crop soil was 249.4 Mg.hm-2, and that in plastic greenhouse and usual vegetable soil was 280.5 Mg.hm-2 and 269.3 Mg.hm-2, respectively, which were only 12.5% and 8.0% higher than that in cereal crop soils. The total-N in plastic greenhouse and usual vegetable soils was 37.5 Mg.hm 2 and 32.7 Mg.hm-2, which was 36.2% and 18.6% higher than that (27.5 Mg.hm-2) in cereal crop soils, respectively. The ammonium-N was 211.5 kg.hm-2 and 197.8 kg.hm 2, which was 29.6% and 21.2% higher than that (163.2 kg.hm-2) in cereal crop soils. The available-K was 6567.8 kg.hm-2 and 5523.6 kg.hm-2, which was 30.6% and 9.8% higher than that (5029.7 kg.hm-2) in cereal crop soils. Furthermore, serious nutrient leaching occurred in vegetable soil profiles due to over-fertilization and irrigation. PMID- 12561169 TI - [Obstacles of soybean continuous cropping, III. Mechanism of soybean yield increment by marine actinomyces MB-97]. AB - An isolate of marine actinomyces MB-97 identified as Streptomyces microflavus could successfully colonize in the rhizoshpere of soybean, and inhibit Penicillium purpurogenum, a soybean deleterious rhizospheric microorganisms. After applied MB-97, the ratio of bacteria/fungi in the rhizosphere of soybean was increased, and the soil became to be "Bacterial type" from "Fungal type". The populations of P. purpurogenum were apparently suppressed about 80%, and the harm of toxins in soil was weak. The soybean root rot caused by soilborne fungi such as Fusarium was decreased 50%, and MB-97 could stimulate the growth of soybean seedlings. In field study, the mean yield of soybean raised by 15.2%, implying that Streptomyces microflavus was an effective plant growth-promoting rhizobacteria on soybean. PMID- 12561171 TI - [Preliminary research on environmental characteristics of oasis-desert ecotone]. AB - Taking Minqin Oasis in the downstream of Shiyanghe Rivers as example, the shrub characteristics of sand dune, which was closely related to the vegetation of the interface between the outside of oasis and desert, was studied in this paper. The change of vegetation coverage and dune density indicated that the vegetation coverage in the ecotone was obviously higher than that in the outside of oasis and in drift desert. There existed an area outside oasis that was obviously affected by human activies and 0-600 m away from the outside of oasis. Meanwhile, the appearing position of area outside oasis was different in different place. The naked area ratio of dune in the area outside oasis was the biggest, so that the area outside oasis was the main resource of drift sand resulting in desertification of oasis. Over extracting ground water in oasis district resulted in a forming of dropping funnel of ground water there, and resulted in a fluid of ground water from desert to oasis. Ultimately, human activites aggravated those processes to some extent. PMID- 12561170 TI - [Response of N transformation related soil enzyme activities to inhibitor applications]. AB - With an aerobic incubation test, this paper studied the response of soil urease, nitrate reductase, nitrite reductase, and hydroxylamine reductase to urease inhibitor hydroquinone (HQ) applied in combination with nitrification inhibitor encapsulated calcium carbide (HQ + ECC) or dicyandiamide (HQ + DCD). The results showed that HQ + DCD could inhibit urease activity and increase activities of nitrate reductase, nitrite reductase, and hydroxylamine reductase significantly in comparison with CK, HQ and HQ + ECC. Under the condition of our test, there existed a significant relationship between soil urease, nitrate reductase, nitrite reductase, and hydroxylamine reductase activities and soil NH4+ and NO3- contents, NH3 volatilization and N2O emission rate, and regression analysis indicated that there were significantly positive relationships between soil urease, nitrite reductase and hydroxylamine reductase activities. PMID- 12561172 TI - [Physiological response of two divergent Leymus chinensis types to drought stress in the Songnen Plain]. AB - The physiological response of two divergent Leymus chinensis types (grey-green type and yellow-green type) to drought stress was studied by pot experiment. At seedling stage, some physiological indexes of the plants and leaves were measured by taking the divergent types under stress of polyethylene glycol (PEG). Quantitative analyses showed that under drought stress, the electrolyte leakage rate, proline content, water content, fresh weight, and SO(4)2- and NO3- contents of two divergent types varied significantly. According to the simulating results by equation y = a + bx, the b values of electrolyte leakage rate and chlorophyll content were higher in grey-green type than in yellow-green type, and the b value of water content of grey-green type was lower than that of yellow-green type. The proline content of grey-green type was higher than that of yellow-green type. Factor analyses showed that proline was the most important factor of physiological response to drought stress of the two types. The two types of L. chinensis had certain capabilities of drought resistance, but the capability of grey-green type was greater than that of yellow-green type. Therefore, the physiological characteristics of drought resistance were significantly differentiated. PMID- 12561173 TI - [Application of SPOT 4 remote sensing imagine in classification of Qira oasis landscape in China]. AB - The landscape surounding Qira osais at the southern fringe of the Taklamakan Desert in China was analyzed by using the SPOT 4 multispectrum remote sensing data and GPS (global positioning system) data. The SPOT 4 scene was projected into an UTM grid, and a supervised classification by ERDAS IMAGINE software was applied. In total, 13 landscape units could be distinguished, and a "frame of reference" was set up for establishing monitoring system of landscape patterns dynamics. Limitations, possible improvements and further applications of the approach were discussed. PMID- 12561174 TI - [Ecological functions of green land system in Harbin]. AB - Patch and strip green lands in Harbin with plantation structure of tree + shrub + herb, tree + herb, shrub + herb, tree, shrub, and lawn were selected as studied objects. Through testing PAR, temperature (t), relative humidity (RH), and CO2 concentration in the center and outside of the selected green lands, the ecological functions of the green lands on regulating city microclimate were analyzed. The results showed that except lawn, patch green lands had the function of shading. Green lands with plantation structure of tree + shrub + herb, tree, tree + herb, and shrub had greater effects on lowering temperature, increasing humidity, and adjusting CO2 concentration than that of lawn. Strip green lands also had certain effects on shading, lowering temperature, increasing humidity, and adjusting CO2 concentration. Among different plantation structures of stripe green lands, shrub had the best effect on shading, and tree + shrub had the best effects on lowering temperature, increasing humidity, and adjusting CO2 concentration. The ecological functions of patch green lands were better than those of strip green lands with the same plantation structure, and green lands with multistoried structure had greater ecological effects than those with single layer. PMID- 12561175 TI - [Dust removal effect of urban tree species in Harbin]. AB - Dust removal by plants was proposed as an important basis of designing urban green space system for future urbanization. Dust removal measurement and leaf surface electron-microscopic scanning were taken for 28 tree species in Harbin city. The results showed that there were obviously different effects of dust removal among different tree species. Picea koraiensis and Juniperus rigida were the ideal conifers for dust removal, and their dust removal abilities were 5.7 g.m-2 and 4.4 g.m-2 after 4 weeks, respectively. Populus alba x berolinensis, Lonicera maackii, and Prunus maackii were the ideal deciduous tree species, and their dust removal abilities were respectively 1.9 g.m-2, 2.0 g.m-2, and 2.45 g.m 2 after 2 weeks. Observation under electronic microscope showed that tree species, having deep channels or dense hair on leaf surface had greater effects of dust removal, while tree species, which had strumose projections on leaf surface, had weaker effects. Different modes of dust removal by plants were also discussed. PMID- 12561176 TI - [Spring propagation and size dynamics characteristics of two kinds of bee populations in Anhui Province]. AB - Systematical observations and researches were conducted on the population size dynamics of Apis mellifera Ligustica Spi. and Apis cerana cerana Feb. in Wanzhong, Wanxi and Wannan mountainous area in Anhui Province in 1997-1999. The results showed that the bee population size was influenced by climate and flower fertility, which was higher in Spring and Autumn, and lower in Summer and Winter. The propagation and renewal of A. mellifera in Autumn were quicker than those of A. cerana cerana, while the effect of overcoming Summer was inferior to that of Apis cerana cerana. The sex ratio of A. mellifera was (314.4 +/- 289.9):1-(329.4 +/- 305.8):1, and that of A. cerana cerana was (334.2 +/- 235.5):1-(413.1 +/- 377.2):1. The birth of drones was seasonal, and the age structure of each bee population was variable. PMID- 12561177 TI - [Theories and methodologies of engineering designs on sustainable agricultural land consolidation project--a case study of Xuemeiyang land consolidation project in Changtai County, Fujian Province]. AB - The concept and characteristics of engineering designs on sustainable agricultural land consolidation project were discussed in this paper. Principles, basic methods and procedures of engineering designs on agricultural land consolidation project were put forward, which were successfully adopted for designing agricultural land consolidation in Xuemeiyang region of Changtai County, including diversity designs of sustainable land use, engineering designs of soil improvement, roads, ditches, and drains for protecting existent animal environments, and design of ecological shelter-forests in farmland. Moreover, from sustainable economic, ecological and social points, the results of these engineering designs were evaluated based on fouteen important indexes. After carrying out these engineeringdesigns, the eco-environments and agricultural production conditions were significantly improved, and the farm income was increased in planned regions. PMID- 12561178 TI - [Bioremediation for petroleum-contaminated soil by composting technology]. AB - With composting technology of off-site bioremediation, the bioremediation of soil contaminated by crude oil from Liaohe Oil Field was studied. 4 treatments units were set, each units being 118.5 cm in length, 65.5 cm in width, and 12.5 cm in height. The results showed that when the soil was contaminated with 5.22 g.100 g 1 TPH, the degradation rate of TPH reached 54.2% after 55 days operation by Phanerochaete chrysosporium. The results also indicated that the major factors that effected the TPH bioremediation results were the content of O2 and CO2, the amounts of microorganism that degraded petroleum hydrocarbons, and the changes of pH in contaminated soil. These factors could directly reflect the effect of composting treatment technology, and be used to optimize the operation conditions of composting technology to reach the best result. The treatment engineering adopted periodical aired means, operated very simple, and costed very cheap, whtch provided a practical technology for the bioremediation of petroleum contaminated soil. PMID- 12561179 TI - [Characteristic of binding forms of arsenic in polluted rice seed and their stability]. AB - The protein-binding forms of As in polluted rice seed and their stability were studied using the methods of Sephadex chromatography. The results showed that As in rice seed was mainly combined with the proteins with apparent molecular weights of 54.5 KD and 5.5 KD. The protein-binding form of As (54.5 KD) was unstable, and was easily decomposed into stable and tiny molecular under the treatment of steaming and digestive enzyme. PMID- 12561180 TI - [Conformation transformation of lead in rhizosphere]. AB - With rhizoboxes, this study dealt with the distribution and conformation transformation of lead in the rhizosphere and non-rhizosphere of wheat and rice grown on red soil. The results showed that the predominant forms of lead were acid dissolved and Carb-Pb (bound to carbonates), Exch-Pb (exchangeable-Pb), and FeMnO-Pb (boud to iron and manganese oxides). Exch-Pb in the rhizosphere of both rice and wheat was much higher than those in the non-rhizosphere, which means that the activation process in the rhizosphere was very strong, and bio-available Pb greatly increased. Various levels of Pb treatment and Pb-Cd interaction also had certain effects on the distribution of lead. In treatment of low Pb concentration, FeMnO-Pb was higher than Exch-Pb, but the contrary result was observed in treatment of high Pb concentration. Both Exch-Pb and FeMnO-Pb in wheat rhizosphere decreased with an increase of Cd. Exch-Pb in rice rhizosphere was correlative to the Cd content in soil. Compared with the treatment in the absence of Cd, the activation of Exch-Pb in rice rhizosphere was weaker in 5 mg Cd.kg-1 treatment, but stronger in 10 mg Cd.kg-1 treatment. PMID- 12561181 TI - [Ecological reconstruction of large opencut coal mine through ectomycorrhizal biotechnology]. AB - Pot experiment with discarded greenish mudstone from large opencut coal mine as test soil was conducted to study the application of ectomycorrhizal fungi in the ecological reconstruction of fragile ecological belt of large opencut coal mine of Huolinhe region, Inner Mongolia. The result showed that after the greenish mudstone was inoculated with ectomycorrhizal fungi, the rates of seedling infection and survival were 1.21-2.78 higher in treatment with no fertilizer application than in that with fertilizer application. The test ectomycorrhizal inoculum 010, A and B had a stronger regulation ability in infertile and low penetration coefficient soil. The ammonification of the mudstone could improve its physical and chemical properties, but shorten the survival duration of seedling on account of the accumulation of ammonia and phosphorus in it. PMID- 12561182 TI - [Effect of different environmental factors on the activities of digestive enzymes and alkaline phosphatase of Macrobrochium nipponense]. AB - The activities of digestive enzymes and alkaline phosphatase from the hepatopancreas of Macrobrochium nipponense were determined under different environmental factors (calcium concentrations 20 mg.L-1, 35 mg.L-1, 60 mg.L-1, 80 mg.L-1, 150 mg.L-1; salinity 7@1000, 14@1000, pH 7.6, 8.8, 9.8). The results showed that higher Ca2+ concentration could enhance the pepsin activity, but inhibit the trysin-like activity in hepatopancreas of M. nipponense. The activities of pepsin, trysin-like, alkaline phosphatase in hepatopancreas of M. nipponense were higher under salinity of 14@1000 than under salinity of 7@1000 and 20@1000. It showed that the activities of digestive enzymes and alkaline phosphatase of shrimp increased gradually with increasing pH value from 7.6 to 9.8. PMID- 12561183 TI - [Oxygen consumption and ammonia-N excretion rates of Chlamys farreri]. AB - The oxygen consumption and ammonia-N excretion rates of Chlamys farreri were studied from April to June, 1999. The results showed that the oxygen consumption and ammonia-N excretion rates of Chlamys farreri were positively correlated with water temperature, and negatively correlated with its body weight under appropriate temperature conditions. Under the test temperature (8-28 degrees C), the oxygen consumption rate ranged from 0.48 to 9.09 mg.g-1.h-1, and the ammonia N excretion rate ranged from 0.05 to 1.01 mg.g-1.h-1. With the raising water temperature, the oxygen consumption rate reached peak value at 23 degrees C, and began to decrease at 28 degrees C. The ammonia-N excretion rate continuously increased from 8 to 28 degrees C. The routine metabolism of Chlamys farreri under treatment was obvious higher than the standard metabolism, and the oxygen consumption and ammonia-N excretion rates increased by 35.8% and 75.9%, respectively. PMID- 12561184 TI - [Thermal tolerance, diel variation of body temperature, and thermal dependence of locomotor performance of hatchling soft-shelled turtles, Trionyx sinensis]. AB - The thermal tolerance, body temperature, and influence of temperature on locomotor performance of hatchling soft-shelled turtles (Trionyx sinensis) were studied under dry and wet conditions, and the selected body temperature of hatchlings was 28.0 and 30.3 degrees C, respectively. Under wet condition, the critical thermal maximum and minimum averaged 40.9 and 7.8 degrees C, respectively. In the environments without thermal gradients, the diel variation of body temperature was highly consistent with the variation of both air and water temperatures, and the body temperature was more directly affected by water temperature than by air temperature, which implied that the physiological thermoregulation of hatchling T. sinensis was very weak. In the environments with thermal gradients, hatchling turtles could maintain relatively high and constant body temperatures, primarily through behavioral thermoregulation. The locomotor performance of hatchling turtles was highly dependent on their body temperature. Within a certain range, the locomotor performance increased with increasing body temperature. In our study, the optimal body temperature for locomotor performance was 31.5 degrees C, under which, the maximum continuous running distance, running distance per minute, and number of stops per minute averaged 1.87 m, 4.92 m.min 1, and 6.2 times.min-1, respectively. The correspondent values at 33.0 degrees C averaged 1.30 m, 4.28 m.min-1, and 7.7 times.min-1, respectively, which indicated that the locomotor performance of hatchling turtles was impaired at 33.0 degrees C. Therefore, extremely high body temperatures might have an adverse effect on locomotor performance of hatchling turtles. PMID- 12561185 TI - [Inhibitory effect of flavones from Ageratum conyzoides on the major pathogens in citrus orchard]. AB - Inter-cropping with Ageratum conyzoides may effectively control major pathogens in citrus orchard. The research showed that flavones produced by and released from A. conyzoides could inhibit the major fungal pathogens, such as Elsimoe fawcetti Bit. Et Jenk, Collectrichum gloeosporimdes Penz, Oidium tingitaninum Carter, Capondium citri Berk et Desm in citrus orchard. 10 flavones, including one glycoside, from A. conyzoides plant were isolated and identified. Most of them could gradually degraded in soil and only 3 flavones could be accumulated and last a long time. Just these 3 flavones could obviously possess inhibitory effect on fungal pathogens in citrus orchard. The inhibitory effect of the 3 flavones was stronger, not only than those of other allelochemicals produced by and released from A. conyzoides, but also than that of Carbenzin, a commercial fungicide. Therefore, the flavones produced by and released from A. conyzoides played an important role in pathogens control in citrus orchard. PMID- 12561186 TI - [Effect of atmospheric CO2 and temperature increment on crop physiology and ecology]. AB - The effect of atmospheric CO2 and temperature increment on crop growth, photosynthesis, yield, and water and nutrient use efficiency was summarized and discussed in this paper. Under high CO2 concentration in the future, the photosynthetic rate of crops may be enhanced to some extent, and their biomass and yield may also be increased. Their stomatal conductance may be reduced, while water use efficiency (WUE) may be increased under elevated CO2 concentration. The aboveground biomass and root production, especially fine root production, will be greatly enhanced, and consequently, litter production increased, C/N ratios increased, plant litter decomposition rates declined under CO2 enrichment. Plants grown under elevated CO2 concentration may release more substrate into rhizosphere, which may lead to an increased activity of soil microorganisms, acclerated mineralization of nutrients, and improved nutritional status of the plant. PMID- 12561187 TI - [Crop productivity model and its application]. AB - Within agricultural ecosystems the interaction between crop and surroundings is quite complicated. In order to know the feedbacks of crop to surroundings in agricultural ecosystem, a research on crop productivity model is becoming important. This paper discussed the backgrounds of the emergency of crop productivity model, thinking that the crop productivity model was a means which could be used to simulate growth process by mathematical ways and computer technology, and analyzed the physiology of crop growth and the correlation between crop and surroundings. At the same time, it discussed four phases of crop productivity model developing, including infancy, youth, adolescent and maturity. Secondly, the paper summarized the crop productivity model's function of agricultural eco-environment protection from three aspects, including scientific research, management of crop, and analysis of agricultural decision, and discussed the disadvantages of crop productivity model. The disadvantages embodied in three facets: 1) though the simple crop productivity model could be used easily in practice, its regional adaptiability was too weak; 2) complicated crop productivity model had too much parameters to obtain, which made it difficult to use in practice; 3) the inconsistency of basic data format in different research areas also made the model's regional adaptability too weak. It's indicated in this paper that the establishment of universal and uniform basic data format is favorable to the extension and application of crop productivity model in other regions. Geographic information system (GIS) was emerged in 1960, which has powerful functions of obtaining, depositing, managing and analyzing spatial data, and is useful to solve the problem of weakly regional adaptability. Meanwhile, friendly interface is favorable to the extension in common users. Finally, the paper discussed the combination of crop productivity model and geographic information system, summarized some researches on friendly interface of crop productivity model, and pointed out that the establishment of universal crop productivity model interface is the focus of the future. PMID- 12561188 TI - [Advances in studies on the effect of surfactant on bioavailability of polycylcic aromatic hydrocarbons (PAHs) in soil]. AB - The solubility and adsorption/desorption equilibrium of PAHs in soil and their interaction with soil bacterium can be altered by surfactants, which lead to the alternation of PAHs bioavailability. The bioavailability of PAHs can be enhanced by the decrease of interface tension between soil and water, the increasement of PAHs' solubility, and the transportation facilitation of PAHs in the presence of surfactants. It also can be inhibited by the surfactant toxicity to the bacteria or by the competitive ultilization between non-toxicitic surfactants and PAHs by the bacteria. In addition, the effects of surfactants on the PAHs of different existence-forms in soils are different. The bioavailability of PAHs can be affected by the type and concentrion of surfactants, PAHs and soil microorganisms, and also by soil physi-chemical characteristics. PMID- 12561189 TI - [Advances in researches of environmental behavior of sulfonylurea herbicides in soil]. AB - In this article, the environmental behavior of sulfonylurea herbicides in soil, which included absorption and de-sorption, degradation, resistance, mobility, and inhibition on soil enzyme activity, was reviewed, and the influences of some soil factors, such as pH, soil type, water content, and organic matter on their environmental behavior were also discussed. Distributed Reactivity Model for Sorption was used to explain the hysteretic behavior of sulfonylurea in soil, and the biotic and abiotic degradation mechanisms were summarized. Some prospects on their ecological toxicity, adsorption strength, and degradation model were also discussed. PMID- 12561190 TI - [River ecosystem health and its assessment]. AB - Ecosystem health is a hot topic in ecological research. In this paper, the concept of river ecosystem health was generalized, and the main methods used to assess the health condition of river ecosystem was introduced, which are based on bioindictors such as epilithic algae (diatom specially), benthic invertebrate and fish, and were classified into two types in terms of their assessment principles. Based on research actuality, the development tendency in this area is put forward. The authors suggest that the relevant study should be carried out as soon as possible in China. PMID- 12561191 TI - [Effect of Pinus sylvestriformis seedlings density on net photosynthetic rate under elevated CO2]. AB - The effect of Pinus sylvestriformis seedlings density on net photosynthetic rate was studied under elevated CO2. Atmospheric CO2 concentration was controlled in OTC (Open Top Chamber). The results showed that elevated CO2 not only made net photosynthetic rates (NPRs) of two Pinus sylvestriformis seedlings densities increased, but also mitigated their intra-specific competition. Meanwhile, the difference of seedling NPRs between 100 and 400 plant.m-2 under 500 mumol.mol-1 air CO2 concentration was less tha that under 350 mumol.mol-1 with the same PAR levels. When air CO2 concentration reached 700 mumol.mol-1, the NPRs of seedlings under both planting densities were close to each other with the same PAR levels. The intra-specific competition was minimized under air CO2 concentration of 700 mumol.mol-1. PMID- 12561192 TI - [Correspondence analysis between occurrence type of Dendrolimus punctatus and topographical and vegetative features]. AB - The relationship between the occurrence type of Dendrolimus punctatus and the geographical and vegetative features in subcompartments was analyzed using correspondence analysis and shown by a figure of cluster. The results showed that the majority subcompartments in which D. punctatus occurred frequently were at low elevation and low slope, and distributed among low hills or middle hills, while the subcompartments at high elevation or high slopes, or in low mountains or middle mountains were safe-type. The subcompartments with only one dominant species, especially Pinus elliottii, were easily attacked by D. punctatus. The subcompartments in conifer and broadleaf mixed forest had less insect pests than those in conifer mixed forest. PMID- 12561193 TI - [Progress in the study of the structure and function of Cre recombinase]. AB - The Cre recombinase, an integrase from bacteriophage P1, catalyzes site-specific recombination between 34-bp repeats termed loxP sites, in the absence of any additional cofactors and energy. Mediated by Cre recombinase, specific DNA fragments can be excised, inversed or integrated depending on the orientation or position of loxP sites in vitro or in vivo. Because of its simplicity and high efficiency, Cre/loxP site-specific recombination system has been widely used in gene deletion and function identification, gene site-specific integration, gene trapping and chromosome engineering. It has been used as a useful tool for DNA recombination in transgenic yeast, plants, insects and mammals. Here progress in the study of the structure and function of Cre recombinase is discussed. PMID- 12561194 TI - [High expression and identification of DNA mismatch repair gene mutS in Escherichia coli]. AB - DNA mismatch repair gene mutS (2.56 kb) was PCR modified and cloned into a secretive prokaryotic expression vector pET32a (+) which carries a N-terminal His.tag + and thioredoxin sequence. MutS protein was expressed with high level after IPTG induction using the strain E. coli AD494(DE3). SDS-PAGE revealed that the expected protein with a molecular weight of 108 kD which is about 35% of the total bacterial proteins is almost soluble. The expected protein was purified directly by immobilized metal (Ni2+) chelation affinity chromatography and the purity is over 90%. MutS protein activity verified using mismatch DNA showed that the expression product can recognize and bind to base-pair mismatch specifically. PMID- 12561195 TI - [Fusion expression of human thymosin alpha 1 in Escherichia coli]. AB - Engineering E. coli strain, BL21 (DE3)/pGEX-4T-human Thymosin alpha 1, was constructed by oligonucleotide annealing and PCR amplifying the target gene, then ligating it with pGEX-4T-3 vector and transferring into BL21 host. The yield of fusion protein of GST-Thymosin alpha 1 expressed from BL21 (DE3)/pGEX-4T-thymosin alpha 1 is about 35%-40% of total protein after fermentation. Following the simple cut of thrombin or CNBr, about 0.2 g/L thymosin alpha 1 can be harvested. The product is checked by MS and activity test, which indicates that the recombinant product has full biological activity of native thymosin alpha 1. PMID- 12561196 TI - [The expression of humanized Fab fragment of the anti-HBsAg antibody in methylotropic yeast Pichia pastoris]. AB - Using of two-step integrating technology, transducted the H and L chain gene of humanized Fab fragment of anti-HB-sAg antibody into the genome of methylotropic yeast P. pastoris. Constructed a engineering yeast to produce humanized Fab fragment of the anti-HBsAg antibody. The Fab fragment was efficiently secreted into the medium at a concentration of 50-80 mg/L. The Fab fragment was purified from culturing supernatant of the recombinant yeas by affinity chromatography. The ELISA analysis showed the high affinity of the expressed humanized Fab fragment to the HBsAg. PMID- 12561197 TI - [Regulation of Gal beta 1,3GalNAc alpha 2,3-sialyltransferase (ST3GalI) by hepatitis B virus MHBst/HBx transactivator]. AB - Hepatitis B virus MHBst and HBx fragments were amplified to construct eukaryotic expression vector pCDNA3.1-MHBst and pCDNA3.1-HBx. ST3GalI promoter region was obtained by the method of PCR and GFP report plasmid pEGFP-N1-Psial was constructed. pCDNA3.1-MHBst or pCDNA3.1-HBx with pEGFP-N1-Psial were transiently co-transfected into QGY-7701 cells using calcium phosphate-DNA co-precipitation, respectively. The expressions of Psial-directed GFP were analyzed by FAC Scalibur. It was found that MHBst/HBx could upregulate ST3GalI promoter activity by 35.2% and 43.8%, respectively. We report the regulation of ST3GalI by MHBst and HBx transactivators. It would be helpful to further investigate the relation between hepatitis B virus infection and sialyltransferase expression. PMID- 12561198 TI - [Screening of TNF-alpha antagonist peptides from a random peptide library displayed with Escherichia coli flagellar]. AB - Tumor necrosis factor(TNF-alpha) plays an improtant role in the process of anti infection and anti-cancer. It can both protect and make damage to the host. In order to find new way of inhibiting its host-damaging activity, An E. coli flagella displayed random peptide library was constructed and TNF-alpha antagonist peptides were screened using the peptide library. After 5 rounds of panning and DNA sequencing, six peptide sequences were obtained. Two of them(TBP2, TBP3) have the same sequence frame of V--N-WG. After primary comparation of TNF-alpha binding ability, four peptides were synthesised and purified with RP-HPLC. The activity of inhibiting TNF-alpha was detected with L929 cell and MTT method. The data show that TBP2 and TBP3 can inhibit 90% of TNF alpha activity when TNF-alpha gives about 30% cell toxicity on L929. The two sequences have not been reported. PMID- 12561199 TI - [Purification and characterization of antifungal proteins in triticale seed]. AB - Using Trichoderma as an indicative fungus, three antifungal proteins in Triticale Zhongsi 237 seed were purified and characterized. These protein components were considered to be a new Class II chitinase and two kinds of beta-1, 3-glucanases. Chitinase molecular mass was 30.5 kD and enzyme activity was maximal at pH 6.0 and 37 degrees C. Two beta-glucanases molecular masses were 51 kD and 23 kD. N terminal amino acid sequences of Triticale chitinase share high homology with barley chitinase. In some conditions, the chitinase and beta-glucanases all had strong antifungal activity and were able to inhibit Trichoderma growth synergistically. Moreover, the chitinase and beta-1, 3-glucanases were able to inhibit powdery mildew growth on detached susceptible wheat leaves. PMID- 12561200 TI - [Cloning and sequencing of lactase gene from Aspergillus candidus]. AB - Genomic DNA and cDNA sequences of lactase from Aspergillus candidus were cloned. Sequences analysis revealed that the genomic DNA was 3458 bp containing eight introns, cDNA was 3015 bp and encoding a polypeptide of 1005 amino acid residues. Signal peptide was 19 amino acid residues, eleven potential N-glycosylation sites were assumed. Comparing the gene cDNA and amino acid sequences with other lactase sequences from various sources, it showed a very low homology with most of other sequences. Although the gene had a higher homology to Aspergillus oryzae lactase sequence, characterization of both enzymes exhibited obvious difference. The gene from Aspergillus candidus was a promising new gene for food industry. PMID- 12561201 TI - [BjCHI1 from Brassica juncea displays both chitinase and agglutination activity]. AB - The proteins encoded by the Brassica juncea chitinase gene BjCHI1 and its derived genes BjCHI2 and BjCHI3 were expressed by Multi-copy Pichia expression system. The chitinase activity of FPLC purified BjCHI1, BjCHI2 and BjCHI3 were tested and the results showed that all the three proteins degraded both CM-chitin-RBV and colloidal chitin. The Km values of BjCHI1, BjCHI2 and BjCHI3 for CM-chitin-RBV were estimated as 0.799 mg/mL, 0.544 mg/mL and 0.793 mg/mL, respectively. When the colloidal chitin was used as substrate, the Km values were 0.281 mg/mL, 0.388 mg/mL and 1.643 mg/mL, respectively, indicating chitin-binding domain can increase affinity of chitinase to insoluble substrate. In the agglutination activity assay, only BjCHI1 shows activity when the protein concentration was more than 33 micrograms/mL, while BjCHI2 and BjCHI3 without agglutination activity even when the concentration was increased as high as 800 micrograms/mL. This means that the two chitin-binding domains in BjCHI1 are essential for agglutination and BjCHI1 is the first protein which shows both chitinase and agglutination activity identified so far in plants. PMID- 12561202 TI - [The analysis of Bacillus thuringiensis vegetative insecticical protein gene cloning and expression]. AB - Three kinds of Bacillus thuringiensis serotype-subsp. Leesis(H33) strain YBT-833, subsp. Aizawai(H7) strain YBT-1416 and subsp. Kurstaki(H3ab) strain YBT-1535, which were isolated by our lab, are chosen as original strain to clone vegetative insecticidal protein gene. Southern hybridization showed that vip genes are all localized at roughly 4-5 kb size-fractionated XbaI fragments of total DNA from YBT-833, YBT-1416 and YBT-1535. Three subgenomic libraries containing the vip gene fragment, were constructed with pUC19 as vector. Then, three vegetative insecticidal protein gene vip83, vip14 and vip15 are obtained from the libraries through the methods of colony-blot-in-situ screening and enzyme-cut detection. Comparision of DNA sequence made out that only vip83 gene exist five different base pairs with known vip genes. Because the sequences of vip14 and vip15 are the same, two of the three genes, vip83 and vip14, were subcloned to shuttle vehicle pHT315 to get recombinant plasmids pBMB8901 and pBMB8902 in turn. The plasmids were separately transformed into vip Bt. receptors BMB171 and 4Q7 to obtain four engineered strains BMB8901-171, BMB8902-171, BMB8901-4Q7 and BMB8902-4Q7. SDS PAGE results indicated that all recombinant strains express 88 kD vegetative insecticidal protein. Bioassay also showed that the proteins of genes vip83 and vip14 both have certain toxicity to Lepidopteran insect larvae such as Heliochis armigera, Spodotera exigua and Plutella xylostella. While the toxicity of vip protein from four engineered strains to Plutella xylostellas are highest, whose LC50 value is 28.6, 31.6, 45.4 and 37.6 microL/mL respectively. This study will contributed to construct high efficacy and wide spectrum engineered strains on theory and reality. PMID- 12561203 TI - [Characterization of inductive synthesis of levoglucosan kinase by a combined strategy of enzymological and fast atom bombardment mass spectrometric analysis]. AB - Levoglucosan is the main product derived from pyrolysis of cellulose. A mutant Aspergillus niger CBX-209 could grow on levoglucosan well fermenting it into citric acid with a yield comparable to that on glucose. Levoglucosan hydrolase was absent by measuring glucose formation with the glucose oxidase and peroxidase coupling system. Cell extracts were partly purified by ammonium sulfate fractionation and ion-exchange chromatograph. Direct formation of glucose 6 phosphate from levoglucosan in the presence of ATP and MgCl2 was observed when it was reacted with partly purified enzyme by a combined strategy of enzymological and fast atom bombardment mass spectrometric analysis. These data showed that the mutant used a novel enzyme, levoglucosan kinase, to convert levoglucosan into glucose 6-phosphate. Levoglucosan kinase was an inductive enzyme. PMID- 12561204 TI - [Design and preparation of the multimeric self-cleavable hammerhead ribozyme targeting apple scar skid viroid and its activity detection in vitro]. AB - A self-cleaving hammerhead ribozyme gene containing a 14nt target sequence of ASSVd at the 3' end of hammerhead ribozyme was synthesized, amplified and cloned at the Xho I-Hind III site of pGEM7Zf(+). The ends produced by Xho I or Sal I can link together, thus the recognition sites of both enzymes vanish and can't be cut by either one. We used this property to get the recombinant plasmid bearing 2, 4, 6, 8, 10 and 12 copies of self-cleavable ribozyme respectively after successively sub-cloning five times. Linearized recombinat plasmid model catalyzed by T7 RNA polymerase was transcribed in vitro. The multimeric ribozyme molecules efficiently self-cleaved via cis-acting to release many ribozyme molecules It indicates that the concentration of ribozyme transcripts has been enhanced during transcription. Trans-cleavage reaction was carried out by incubating monomeric and multimeric ribozymes with same mol concentration and 32P labeled target ASSVd. Both ribozymes and target transcripts were mixed in 1:1 ratio. Autoradiograms showed the transcripts of multimeric ribozyme were substantially more effective against the ASSVd target RNA than the monomeric ribozymes. We confer that the multimeric self-clevable ribozyme is likely to provide more valuable application in vivo. PMID- 12561205 TI - [Cloning and expression of kringle 1-3 gene of human plasminogen and the purification and bioactivity of its expressed product]. AB - Kringle 1-3 domain is a recently found angiogenesis inhibitor with anti angiogenesis and anti-tumor activity. The kringle 1-3 gene was amplified by PCR technique using angiostatin gene as template. After DNA sequencing, the PCR product was cloned into pPIC9K resulting in recombinant plasmid pPIC9K13 which was then transformed into Pichia pastoris GS115. The high copy integration transformants screened by PCR and G418 methods were cultivated in flasks. The K1 3 was expressed and secreted to the medium and has immunogenic activity as shown by SDS-PAGE and Western blotting. High cell density culture was carried out in 30 liter and 80-liter bioreactor, the biomass reaches 300 OD after methanol induction, and the expressed product is 200 mg/L. The fermentation supernatant was purified by Streamline SP and Phenyl Sepharose Chromatography, the product appears as a single band on SDS-PAGE, with a purity of 95%-96%. The purified product has anti-angiogenesis and anti-tumor activity. PMID- 12561206 TI - [Cloning of staphylococcal enterotoxin B gene and its highly expression in Escherichia coli]. AB - An about 700 bp DNA fragment was amplified from genome DNA of S. aureus TSTw by PCR. This fragment was cloned into pGEM-7Zf(+) and the recombinant plasmid was transformed into E. coli DH5 alpha. The sequencing result of the recombinant plasmid demonstrated that it contains seb gene with 717 bp (without signal encoding region of 81 bp) which has the same nucleotide sequence as described in literature. The seb gene was cloned into expression vector 7ZTS and was transformed into E. coli JM109 (DE3). The expression level of SEB was as high as 33.3% of the cell total proteins. PMID- 12561207 TI - [Induction of immune responses in mice by hepatitis B virus large envelope DNA vaccine delivered by attenuated Salmonella typhimurium]. AB - The enhanced green fluorescent protein (EGFP) expression plasmid was transformed into an attenuated AroA- autotrophic mutant of Salmonella typhimurium SL7207, the resultant bacteria was administered orally to BALB/c mice. EGFP expressed in spleen cells was detected by flow cytometry. A DNA vaccine encoding HBV large envelope protein was immunized BALB/c mice by oral delivery through SL7207 or by direct intramuscular injection. The serum antibodies, T lymphocyte proliferative response and cytotoxic T lymphocyte response of mice were detected. The results showed that both DNA immunization methods could induce cellular and humoral immune responses, whereas oral vaccination elicited stronger immune responses than intramuscular vaccination did. Therefor, oral administration with HBV DNA vaccine using attenuated Salmonella may be a simple and effective method for the therapy of hepatitis B. PMID- 12561208 TI - [Cloning of the major antigen region of E2 gene of hog cholera virus and expression in Escherichia coli]. AB - The major antigen region of E2 gene of Hog Cholera Prevalent Strain (Guangxi Yuling Strain) and Chinese Hog Cholera Lapinised Virus (C-strain) derived from hog and rabbit spleen tissue, was amplified by reverse transcription polymerase chain reaction(RT-PCR) and the nested Polymerase Chain Reaction (nPCR). After the amplified fragments were cloned into the expression vector pPROEX-HTb, the recombinant plasmids pPROEX-GXYL and pPROEX-C were obtained. The insert position, the size and the reading frame were right by PCR, restriction digestion and the sequence analysis. SDS-PAGE indicated that both of the reciepient germs transducted and induced by the recombinant plasmids pPROEX-GXYL and pPROEX-C could express the major antigen region of E2 gene. Western-blot indicated that the expressed antigen protein could be recognized by the positive serum of CSFV. PMID- 12561209 TI - [Preparation of bovine lipid-free hemoglobin]. AB - A new method for preparation of Hb solution free of stromal lipid was described. Almost all the lipid in fresh hemolysate of bovine red blood was removed with hydrophobic interaction chromatography (HIC) in the presence of 2% PEG4000, 5% PEG4000, 2%PEG10000 or 5%PEG10000. With the adding of 5%PEG4000, the 80% of recovery of Hb in HIC was obtained and the maximum lipid absorbed by hydrophobic medium, butyl agarose -6B was 86.6 mg/mL. The activity (P50) of hemoglobin preparation was 3386.4 Pa torrs, and the Hill number was 2.54, which were near to that of the native red blood cells. The mechanism of removing lipid by HIC and the function of PEG in the process were discussed. PMID- 12561210 TI - [Biosynthesis and accumulation of poly(3-hydroxybutyrate) in Vibrio natriegens]. AB - Accumulation of poly(3-hydroxybutyrate) [poly(3HB)] by V. natriegens was studied. Results indicated that V. natriegens used glucose, gluconate, fructose and molasses as carbon sources for poly(3HB) synthesis. When molasses was used, up to 28.4% of poly(3HB) to cellular dry weight was accumulated. The accumulation of poly(3HB) followed, was not simultaneously to, the cell growth. Analysis of the PHA polymerase, beta-ketothiolase, and acetoacetyl-CoA reductase showed that the poly(3HB) accumulation was correlated to the increase of their activities in cells. Poly(3HB) accumulation was also related to the de novo fatty acid synthesis, as revealed by the results that cerulenin, a specific inhibitor to the de novo fatty acid synthesis, significantly reduced accumulation of poly(3HB). Based on the results from this study, the synthetic pathway of poly(3HB) was proposed. PMID- 12561211 TI - [Micropatterning of biotin-avidin layers and cell location]. AB - The located culture of cells on patterned surfaces is useful for tissue engineering, biosensor development and fundamental research of cell biology. It is presented here a rapid fabrication method of Biotin-Avidin protein layers micropattern, which is based on soft-lithography technology. The bovine aortic endothelial cells are cultured on the micropatterned surface. It is found that cell location can be controlled on the scale of individual cell by this method. PMID- 12561212 TI - [Analysis on metabolic flux shift during guanosine fermentation]. AB - Taking the typical metabolic control product-guanosine as an example, the method of metabolic flux shift investigation based on process multi-levels parameter correlation analysis was established. The metabolic pathway, multi-parameter correlation, accumulation of amino acid and organic acid during guanosine fermentation process were integratively analyzed. The metabolic flux shift from HMP to EMP was ascertained, which was assumed to be caused by the accumulation of ammonium ion. The subsequent optimization based on controlling flux distribution between EMP and HMP did improve the yield by 35% when the metabolic flux shift was prevented. PMID- 12561213 TI - [Calcium-regulated growth and differentiation of the mouse epidermal keratinocytes]. AB - The effect of the calcium ion (Ca2+) on the growth and differentiation of the mouse epidermal keratinocytes cultured in serum-free medium was investigated. It was found that the optimal level of calcium ion in the medium was about 0.2 mmol/L. Under such a culture condition the colony forming efficiency, attachment percentage, percentage of the cells with cornified envelops, and percentage of the senesced cells were measured to be about 10.8%, 30.8%, 5.1%, and 26.8%, respectively. However, the Ca2+ concentrations in the medium above 0.6 mmol/L resulted in significant differentiation and senescence of the keratinocytes, which was found to be harmful for keratinocyte growth and expansion in vitro. PMID- 12561214 TI - [The effect of antioxidants on the in vitro life-span of keratinocyte]. AB - The effect of antioxidants on the in vitro life span of mouse keratinocytes was investigated in this work. It was found that the life span of the keratinocytes cultured in the medium supplemented with antioxidants was extended significantly. The most beneficial antioxidant used in this work was the mercaptoethanol, followed by the catalase and SOD. However, the growth rates of keratinocytes in vitro under all the experimental conditions still declined with the culture time. It was also found that the antioxidants added in the medium were also helpful to enhance the keratinocyte colony formation. In addition, the aging kinetics of the mouse epidermal keratinocytes in vitro were analyzed, and finally the aging rate constants corresponding to antioxidants used were calculated. PMID- 12561215 TI - [Influence of pH control on the production of curdlan by Alcaligenes faecalis strain]. AB - A two-stage pH control method was employed in batch fermentation of curdlan by Alcaligenes faecalis WX-C12 where cell-growth stage was constantly controlled at pH 7.0 and stationary stage was controlled at a constant pH as well. The influence of pH control on the curdlan production was investigated. The optimal pH control of batch process for curdlan production was obtained when cell-growth stage was controlled at pH 7.0 and stationary stage was constantly controlled at pH 5.6. Production and productivity of curdlan, QP and YP/S reached 28.19 g/L, 291 mg/(L.h), 132.27 mg/(L.h.g) and 0.659, an improvement of 20.4%, 38.1%, 38.1% and 29.5% compared to a pH uncontrolled operation respectively. PMID- 12561216 TI - [Induction of petal-like structures from petals of Crocus sativus L. and the differentiation of style-stigma-like structures in vitro]. AB - Firstly the petal of Crocus sativus L was cultured on the medium that supplemented with different combinations of hormones. The petal-like structures(PLS) were induced on medium, but the induction rates were different in various medium. The highest induction rate of petal-like structures was obtained on the media that was supplemented with NAA (4 mg/L) and KT (8 mg/L). The petal like structures were subcultured on another media when the structure was produced on the explants and proliferate groups. The later media was used for inducing style-stigma-like structures(SSLS). The induction rate of style-stigma-like structures in the petal-like structures group is much higher than the rate in the preceding work, and the maximum of style-stigma-like structures produced per explant was 30. The best result of style-stigma-like structures was observed on the petal-like structure groups which came from the third treatment. The differentiation rate of style-stigma-like structures is stable in the subcultures of petal-like structures. The result revealed that the induction frequency of style-stigma-like structures formed on the petal-like structures is higher than that form on the petals of C. sativus L. PMID- 12561217 TI - [Amplification, cloning and sequence analysis of spider dragline silk cDNA]. AB - Spider dragline silk is synthesized in special gland named major ampulate (MA) gland. The MA glands were dissected from the abdomen of the spiders Nephila clavata and the total RNA was extracted by the TRIZOL. The cDNA of dragline silk was amplificated by RT-PCR (reverse transcription polymerase chain reaction), multiplex PCR and cloned. PCR identification, restriction analysis and DNA sequence analysis were carried out to verify the recombinant plasmids. The codon usage frequencies of the cloned cDNA were added up, and the predicted amino acid sequence was compared with Spidroin2 of Nephila clavipes. Predicted secondary structure of the predicted amino-acid sequence was analysized by DNAStar software. All results showed that the cloned cDNA we got (GenBank Accession No. AF441245) was the very fragment of spider dragline silk Spidroin2 cDNA. PMID- 12561218 TI - [The PTD domain of Tat protein enhance GFP protein delivering into myeloma cell SP2/0]. AB - In order to detect the protein delivery mediated by the PTD (protein transduction domain) of TAT Protein, a expression vector, named pT7460-GFP, was constructed by insert the PTD DNA Sequence, followed by a GFP (green fluorescent protein) gene fused in-frame, into the pT7450 vector. The TAT-GFP fusion protein was expressed in the E. coli ER2566. Most of the fusion protein was presented in the inclusion body. The protein was purified by Ni2+ affinity chromatography under denature conditions, then by a Sepharose Q column to remove urea. The soluble denatured protein was added directly to medium containing the Myeloma Cell SP2/0. It came out that the fusion protein could be detected delivered into the cells under fluorescent microscope in a short time. PMID- 12561219 TI - [Molecular cytogenetic detection of minute chromosomal structural abnormality on the chromosomal terminal regions]. AB - In order to identify those easily overlooked minute chromosomal structural abnormality on the chromosomal regions, and to provide a valuable guidance for pregnancy, fluorescence in situ hybridization (FISH) technique by whole chromosome 7 painting probe, Xq probe and subterminal probe of 7q36-->qter was performed to analyze two cases. Case 1 had a history of recurrence spontaneous abotion and with an uncertain minute translocation on the chromosomal terminal regions. Case 2 was a premature ovarian failure patient with a balanced translocation between chromosome X and chromosome 7 by G banding. The results showed that case 1 was a cryptic minute translocation on the chromosomal terminal regions, and the breakpoint of case 2 was accurately determined, that is, the breakpoint was not on 7q36 but on 7qter. Therefore FISH technique with whole chromosome painting probe and subterminal probe could be used to diagnose the minute chromosomal structural abnormality on the chromosomal regions. It could be used widely in the clinical genetics and was an effective tool for genetic counseling and reproductive guidance. PMID- 12561220 TI - [Association between apolipoprotein E gene polymorphism and the patients with persistent vegetative state in the Chinese]. AB - We studied the relationship between apolipoprotein E gene polymorphism and persistent vegetative state (PVS) to explore the genetics background of PVS, and evaluated the effect of ApoE gene polymorphism on lipid levels in plasma. The ApoE genotype of fifty-six patients with PVS and fifty-three controls were determined by PCR and restriction fragment length polymorphism (PCR-RFLP). Plasma lipid levels were measured by using routine methods. Results demonstrated that there were five genotypes in the two groups: E3/3, E3/4, E2/2, E2/3 and E2/4. The genotype frequencies of ApoE gene in PVS were 21(37.5%), 26(46.4%), 2 (3.6%), 5(8.9%), 2(3.6%) and that in control were 37(69.8%), 7(13.2%), 2(3.8%) 5(9.4%), 2(3.8%) respectively. We compared the genotype frequencies between the two groups and found there was a significantly increase in E3/4(chi 2 = 14.236, P < 0.001) and decrease in E3/3(chi 2 = 5.348, P < 0.05). epsilon 2, epsilon 3 and epsilon 4 allele frequencies of ApoE were 11(9.8%), 73(65.2%), 28(25%) in PVS and were 11 (10.4%), 86(81.1%), 9(8.5%) in control respectively. Allele frequencies, significantly increased in epsilon 4 (chi 2 = 10.533, P < 0.001) and decreased in epsilon 3 (chi 2 = 7.022, P < 0.01). We also found that E3/4, E2/4 genotype and epsilon 4 allele can largely increase total cholesterol (TC) and lower density lipoprotein cholesterol (LDL-C) levels in plasma, and epsilon 2 alleles also can largely increase LDL-C levels inplasma. Our finding indicates that the ApoE gene polymorphism may be in association with the PVS, and may be a factor in the genetic susceptibility to PVS in Chinese; Genotype and alleles of ApoE in PVS can affect the lipid levels in plasma. PMID- 12561221 TI - [STR polymorphisms in four Xinjiang ethnic groups in China]. AB - Population genetic studies were performed in Xinjiang's Uygur, Sibe, Ozbek and Kirgiz. Allele frequency distributions were analyzed for ten loci, i.e., D3S1358, VWA, CSF1PO, FGA, THO1, TPOX, D5S818, D13S317 and D7S820 by GeneScan. The results showed that there were 66 STR alleles and 148 genotypes in Uygur; 72 STR alleles and 163 genotypes in Sibe; 65 STR alleles and 168 genotypes in Ozbek; 71 STR alleles and 191 genotypes in Kirgiz. Significant differences were identified among ethnic groups (African-American, US-Caucasian and Chinese-Oriental), but similarity was found among the four Xinjiang population. These findings indicated that the nine STR loci and Amelogenin locus were very useful for individual identification in forensic science. It is further certified that Chinese nationality is a whole. PMID- 12561222 TI - [Molecular cloning and analysis of bovine prolactin full-long genomic as well as cDNA sequences]. AB - In this report, a full-length sequence of bovine prolactin (bPRL) genomic DNA with 9388 bp, which has been accepted by GenBank (Accession Number: AF426315), was firstly cloned by Long PCR procedures. This sequence consists of 5 exons, 4 introns, 854 bp of 5' upstream regulatory region and 69 bp of 3' UTR. Accession number of protein encoded by AF426315 sequence in GenBank is AAL28075 that is composed of 229 amino acid residues, in which signal sequence resides in 1-30 sites and mature polypeptide consists of 199 amino acid residues. The recombinant plasmid containing bPRL genomic DNA was then transfected into eukaryotic cells (COS-7), followed by RT-PCR procedure. An 804 bp of bPRL cDNA containing all the encoding region was obtained, indicating that the bPRL genomic DNA reported herein had its biological function at the transcriptional level. Results derived from information searching by Blast program revealed that there were various SNP sites in the sequences of bPRL mRNAs and ESTs collected in GenBank, which located mainly in downstream encoding region and 3' UTR. These SNP sites did not alter the related amino acids encoded. In addition, mRNA sequences encoding 5' signal sequence of bPRL gene was highly conserve. PMID- 12561223 TI - [Studies of population genetic relationships among 24 Chinese and exotic pig breeds using AFLP analysis]. AB - A total of 12 AFLP primer combinations were used to detect genetic variation of pooled DNA in a sample of 19 Chinese native pig breeds, 1 cultivated pig breed and 4 European and American pig breeds. The genetic similarity coefficient of 24 pig beeds was calculated from AFLP data, UPGMA cluster analysis was also performed. The 12 primer combinations generated more than 1000 bands, of which 208 bands were polymorphic, 17.3 polymorphic markers were detected by one primer combination on the average. Thirteen putative breed specific bands were produced in the pooled DNA of 8 pig breeds. The cultivated pig breed and 4 exotic pig breeds were clustere into one group, while 19 Chinese native pig breeds were gathered into the other group in the UPGMA tree. The result indicated that AFLP analysis had high assay efficiency index (Ai) and provided a valuable tool for assaying genetic diversity and breed characterization in pigs. Chinese native pig breeds and exotic pig breeds show remarkable genetic differentiation, which had farther genetic relationships. Nanchang White pig and Large White pig, Yushan Black pig and Yanshan Black pig had intimate genetic relationships with each other respectively, which were consistent with its breeding history, geographical distribution and RAPD analysis results. In addition, the reasons for cluster results of some pig breeds from AFLP data were not consistent with morphology, geographical distribution and existing classification were discussed. PMID- 12561224 TI - [Molecular cloning of the DNA sequence of activin beta A subunit gene mature peptides from panda and related species and its application in the research of phylogeny and taxonomy]. AB - Activin, which is included in the transforming growth factor-beta (TGF beta) superfamily of proteins and receptors, is known to have broad-ranging effects in the creatures. The mature peptide of beta A subunit of this gene, one of the most highly conserved sequence, can elevate the basal secretion of follicle stimulating hormone (FSH) in the pituitary and FSH is pivotal to organism's reproduction. Reproduction block is one of the main reasons which cause giant panda to extinct. The sequence of Activin beta A subunit gene mature peptides has been successfully amplified from giant panda, red panda and malayan sun bear's genomic DNA by using polymerase chain reaction (PCR) with a pair of degenerate primers. The PCR products were cloned into the vector pBlueScript+ of Esherichia coli. Sequence analysis of Activin beta A subunit gene mature peptides shows that the length of this gene segment is the same (359 bp) and there is no intron in all three species. The sequence encodes a peptide of 119 amino acid residues. The homology comparison demonstrates 93.9% DNA homology and 99% homology in amino acid among these three species. Both GenBank blast search result and restriction enzyme map reveal that the sequences of Activin beta A subunit gene mature peptides of different species are highly conserved during the evolution process. Phylogeny analysis is performed with PHYLIP software package. A consistent phylogeny tree has been drawn with three different methods. The software analysis outcome accords with the academic view that giant panda has a closer relationship to the malayan sun bear than the red panda. Giant panda should be grouped into the bear family (Uersidae) with the malayan sun bear. As to the red panda, it would be better that this animal be grouped into the unique family (red panda family) because of great difference between the red panda and the bears (Uersidae). PMID- 12561225 TI - [Genetic diversity analysis of mitochondrial D-loop region of Chinese sucker (Myxocyprinus asiaticus)]. AB - Chinese sucker, Myxocyprinus asiaticus, is an endemic species of China and also the only representatives of family Catostomidae in Asia. The fish was naturally distributed in Yangtze River and Mingjiang River and now few could be captured because of pollution and overexploitation. The fish has been listed in the second class of preserved animal in China. Studying and assessing its population structure is an imperative and fundamental work for making effective protection strategies. We amplified and sequenced the D-loop region of mtDNA of 8 samples. The size of the D-loop region is about 958 bp. A total of 32 variation loci were detected and the mutation rate was 0.033. All the mutation came from nucleotide substitution except one nucleotide deletion. Most of the nucleotide variations were found between the 55-365 bp region. The individual mutation rate varied from 0-1.36%, which exhibited nucleotide polymorphism to some extent among 8 samples. Compared with RAPD and other PCR-based methods, the directily sequencing of mtDNA D-loop region revealed much more genetic diversity. Meanwhile, the D-loop region of Moxostoma robustum derived from GenBank was aligned with that of Chinese sucker through CLUSTAL software. By comparison, we found that the mutation rate (0.033) of D-loop of Chinese sucker is higher than that of Moxostoma robustum (0.016). PMID- 12561226 TI - [Molecular mapping of QTLs for rice milling yield traits]. AB - QTLs of three milling yield traits, including brown rice (BR,%), milled rice (MR,%) and head milled rice (HR,%), were mapped using a set of 212 Lemont/Teqing RI population, an RFLP linkage map with 182 markers and a mixed model approach(QTLMapper V1.0). The population showed continuous distributions with transgressive segregation on both sides for all traits while HR had a wider variation than BR and MR. One and four main effect QTLs were detected for MR and HR. Two QTLs for HR(QHr6 and QHr7) had large additive effects. Twelve, five and sixteen pairs of digenic epistatic loci were associated with BR, MR and HR respectively. Epistasis was more important than main effect QTLs according to the mapping result. A complex relationship was observed for epistatic pairs mapped in same trait or among different traits by sharing intervals. PMID- 12561227 TI - [Mapping of two fertility-restoring gene for WA cytoplasmic male sterility in minghui63 using SSR markers]. AB - F2 population derived from Shanyou63, F1 hybrid developed from the cross Zhenshan97 A/Zhenshan97B, was used in this study. Fertile bulk was constructed by polling equal amount of 15 highly fertile lines. Sterile bulk was obtained by pooling equal amount of 15 highly sterile lines. Minghui63 and Zhenshan97A, parents of Shanyou63, were analyzed with 302 pairs of SSR primers. 244 pairs of primers gave amplification products, of which 58 pairs of primers on 12 different chromosomes showed polymorphism between the two parents with polymorphic frequency up to 23.77%. Gene bulks were further assayed with the 5 pairs of primers. RM1 on chromosome 1 and RM258, RM304 on chromosome 10 was found to be polymorphic between the two gene bulks. In theory, there should be no difference detected between the two gene bulks except for the target traits governed by fertility-restoring genes. RM1, RM258 and RM304 were probably related to the restorer genes. Ten highly fertile and ten highly sterile lines were selected from F2 population of Shanyou63 to screen the gene bulks. The results showed that specific bands of Minghui63 were detected in all ten highly fertile lines while not observed in all the sterile lines. It indicated that the three SSR markers might be linked to fertility-restoring genes. Dominant lines were not selected due to their inalbility to distinguish recombinant lines from non-recombinant lines. Pure recessive lines were chosen to conduct mapping analysis. A total of 53 highly sterile lines were selected from 900 lines of Shanyou63 F2 population to estimate the genetic distance between three SSR markers and fertility restoring genes respectively. The results demonstrated that recombination occurred in 2, 3, lines with RM1 and RM258 while no one with RM304. Using MAPMAKER/EXP 3.0, the genetic distance between RM1, RM258, RM304 and the related restorer genes were calculated as 1.9, 2.9 and 0.0 cM, respectively. It is possible that the fertility restoring gene(s) on chromosome 10 for three different types of cytoplasmic male sterility(WA, BT and HL) are of the same, or belong to a gene family. PMID- 12561228 TI - [Genetic analysis of blast resistance in japonica rice landrace heikezijing from Taihu region]. AB - Japonica rice landrace Heikezijing (HKZJ) from Taihu region is highly resistant to several Chinese and Japanese differential strains of Magnaporthe grisea. The F1, F2 and RIL populations from the cross between the resistant variety Heikezijing and the susceptible variety Lijiangxintuanheigu (LJXTHG) were inoculated by spray with two strains of Ken 54-04 and Hoku 1 in seedling stages. Based on the R:S ratios of segregation in F1, F2 and RIL populations it was showed that there were two independent dominant genes in Heikezijing in responsible for resistance to strain Ken 54-04 and one dominant R gene to strain Hoku 1 which is the same to one of the two genes resistant to Ken 54-04. The allelic test indicated that the gene with resistance to both Hoku 1 and Ken 54-04 is non-allelic to loci of Pi-k, Pi-z, Pi-ta, Pi-b and Pi-t, also neither Pi-i nor Pi-a gene. It is necessary to confirm whether it is an unknown gene. PMID- 12561229 TI - [Cloning and sequencing of fragments associated with cytoplasm male sterility of rice]. AB - Mitochorndrial DNA (mtDNA), nuclear DNA (nDNA), and genomic DNA (gDNA) were individually extracted from Zhenshan 97A, Xieqingzao A and A-23, which were cytoplasmic male sterile (wild abortive, dwarf abortive and BT), and from Zhenshan 97B, Xieqingzao B and BT, which were maintainers lines. Each of them was analyzed with 500 random primers by RAPD. Six fragments specific to male sterile lines were amplified. A fragment, PWH-17, associated with wild and dwarf abortive type cytoplasmic male sterility was analyzed by Southern blotting and sequencing, and also tested by SCAR. It was 1879 bp in length, which contained 6 open reading frames and 8 repeated sequences. BLAST search revealed that partial sequence in PWH-17 showed high similarity to the upstream sequence of tRNA-Asp gene in Elytrigia elongate (GenBank accession number U14335) and wheat (X13243). The similarity ratios were 97% and 100%. It was inferred that the possible position of PWH-17 in rice (wild abortive) could be located between tRNA-Asp and Cox II, which will be discussed further. PMID- 12561230 TI - [cDNA libraries construction and screening in gene expression profiling of disease resistance in wheat]. AB - A wheat line, Bai Nong 3217/Mardler BC5F4 with resistance to powdery mildew, was used to construct a conventional cDNA library and a suppression subtractive hybridization (SSH) cDNA library from wheat leaves inoculated by Erysiphe graminis DC. Three hundred and eighty-seven non-redundant ESTs from the conventional cDNA library and 760 ESTs from the SSH cDNA library were obtained, and the ESTs similarity analysis using BLASTn and BLASTx were conducted by comparing these ESTs with sequences in GenBank. The results showed that the redundancy of some kinds of genes such as photosynthesis related genes and ribosome related genes was higher in the conventional cDNA library but the varieties and quantities of disease resistance genes were less than those in the SSH cDNA library. The SSH cDNA library was found to have obvious advantages in gene expression profiling of disease resistance such as simple library construction procedure, rich specific DRR (disease-resistance-related) genes and decreased sequencing amount. To acquire genes that were involved in the powdery mildew resistance of wheat, hybridization with high-density dots membranes was used to screen the two libraries. The result showed that the method was relatively simple in operation, and the membranes could be used for many times. But some problems also existed with this screening method. For instance, a large amount of mRNA and radioactive isotope were needed and the hybridization procedure must be repeated several times to obtain stable hybridization results. About 54.1% function-known ESTs in the SSH cDNA library were identified to be DRR genes by screening. There were 247 clones of the SSH cDNA library that had positive signal in the repeated hybridizations with the pathogen uninfected probe. The identified DRR genes distributed in the whole procedure of powdery mildew resistance, but mainly focused on the SAR (systemic of acquired resistance). PMID- 12561231 TI - [Genetic analysis and molecular marker of Avr-Pi1, Avr-Pi2 and Avr-Pi4a of Magnaporthe grisea]. AB - The isolate 81278ZB15 is a fertile strain of predominant virulence of Magnaporthe grisea in Fujian province, China. Its rep-PCR (repetitive element-based polymerase chain reaction) DNA fingerprint and virulence spectrum are different from those of GUY11, a hermophrodite. Ascospores of the cross 81278ZB15 and GUY11 were randomly isolated and their virulence was tested on CO39 NILs (near isogenic lines, C101LAC Pi-1, C101A51 Pi-2, C104PKT Pi-3, C101PKT Pi-4a, C105TTP-4L-23 Pi 4b and CO39). The results showed that 81278ZB15 has Avr-Pil, Avr-Pi2, Avr-Pi4a and Avr-Pi4b genes. The segregation of avirulence and virulence on Pi-1, Pi-2, Pi 4a in the progeny fits 1:1, but there were 8 recombinant ascospore progeny in these three avirulence loci. The results suggest that Avr-Pil, Avr-Pi2 and Avr Pi4a in 81278ZB15 are three tightly linked but independent loci. A 1.2 kb DNA fragment based on rep-PCR fingerprint (RPF1.2) was found to link to these avirulence loci. The genetic distance between RPF1.2 and Avr-Pil, Avr-Pi2, and Avr-Pi4a is estimated to be 5.9 cM, 2.2 cM and 2.2 cM respectively. Furthermore, some progenies are avirulent on Pi-3, CO39 and 8 prognies are virulent on Pi-4b although both parents are virulent on Pi-3, CO39 and avirulent on Pi-4b. The possible causes of irregular recombination were also discussed. PMID- 12561232 TI - [Major-polygene effect analysis of super quality fiber properties in upland cotton (G. hirsutum L.)]. AB - The modern textile industry depends on the improvement of fiber quality, especially strength to meet the needs of higher spinning speed. Inheritance of super quality fiber properties in Upland cotton was conducted in the present paper. P1, P2, F1, B1, B2 and F2 of eight crosses from five parents with different fiber strength, i.e. 7235 x TM1, TM1 x 7235, HS42 x TM1, PD69 x TM1, MD51 x TM1, 7235 x HS42, 7235 x PD69 and HS42 x PD69, and F2:3 for 7235 x TM1, were used in the study. The materials were planted in Nanjing or Hainan in 1998 and 1999, the individual plant fiber samples were tested with HVI system in Cotton Research Institute of CAAS at Anyang. The segregation analysis methods for major genes plus polygene mixed inheritance model developed by Gai were used to identify the genetic system of fiber qualities. The results from joint analyses of multiple segregating generations as well as single segregating generations, especially for F2:3, showed one major gene plus polygene mixed inheritance model in all fiber quality characters. The heritability values of major gene in F2 of 7235 x TM1 with great parent difference were estimated as 19.6% for fiber strength, 32.0% for micronaire and 13.9% for fiber length, but little in B1 and B2 for fiber qualities. The fiber length showed high and positive dominant effect, but negative value or zero of major or polygene dominant effects for other fiber qualities. Therefore, Mid-parent value or tendency to lower parent in F1 for most of fiber qualities lead to low selection efficiency, which suggests that molecular assisted selection should be considered at first in the improvement of fiber qualities. PMID- 12561233 TI - [Analysis of genetic effects for growth traits of Eucalyptus globulus Labill. in a 6 x 6 diallel design]. AB - The genetic effects of general and specific combining ability, male, female, and specific reciprocal cross were estimated for height, diameter of breast height (dbh), volume, crown-width, under branch height, crook degree and ratio of branch with shoot in Eucalyptus globulus Labill. Three quantitative characters, i.e. tree height, dbh, volume were focused, and their effects were discussed in detail. It was found that the GCA, SCA and specific reciprocal cross effects (SRE) were most important, and female effect (FEM) and male effect (MAL) were less important. Three parents with high GCA were good materials used for establishing seed orchard. Eight combinations significantly improved quantitative characters. One of them was improved on quantitative character and trunk quality at the same time. The breeding strategy and methods were also suggested for E. globulus. PMID- 12561234 TI - A general frame of theory on genetic load. AB - Genetic load expresses the loss in mean fitness of a population because of the genetic variability present. The quantitative discussion of various genetic loads is provided with important meaning for research into evolution of species on realistic level. The past theory on genetic load starts off with equilibrium of population to investigate the evolution of species on realistic level. However, the evolution is a displacement on equilibrium of population. This is just made up of a contradiction between theory and praxis. We branch out the past theory on genetic load, and give out a general theoretic frame describing the various genetic loads. By the use of this theoretic frame the genetic loads in an equilibrium population can be represented, and the genetic loads in a non equilibrium population and their change can also be simulated. Thus this theoretic frame overcomes the shortcoming of past theory on genetic loads, which can't describe non-equilibrium population and conflicts with praxis of biological evolution sometimes, and offers a sort of credible simulation methods for research into evolution of species. PMID- 12561235 TI - Formulation development of epidermal growth factor. AB - The information in this paper is presented to summarise various investigations in recent years to develop epidermal growth factor (EGF) delivery systems. EGF is a promising and well-characterised polypeptide that can be used in the treatment of various types of wounds and ulcers. Many approaches have been employed to deliver EGF as a stimulant for cellular activities involved in the processes of wound healing and tissue repair. Recently, emphasis is placed on the development of sustained or controlled delivery technology to optimise EGF delivery. PMID- 12561236 TI - Piperidine-containing histamine H3 receptor antagonists of the carbamate series: the influence of the additional ether functionality. AB - Recently novel leads for histamine H3 receptor antagonists of the non-imidazole type have been described. As a continuation of this research eleven new carbamate derivatives possessing an additional ether functionality were prepared. The compounds were evaluated in vitro for their antagonist activity on isolated organs of guinea-pig (GP) H3 as well as H2, H1, and M3 receptors, respectively. All compounds investigated possessed moderate antagonist affinities at guinea-pig histamine H3 receptors (pA2 6.11-6.76). An ether functionality introduced in different places of the lipophilic part of carbamates differently influenced activity and selectivity toward H3, M3, and other histamine receptors tested. PMID- 12561237 TI - Synthesis of new 2-substituted-[1,3,4]-oxadiazino-[6,5-b]-indoles with H1 antihistaminic, antimuscarinic and antimicrobial activity [corrected]. AB - New 2-substituted-[1,3,4]-oxadiazino-[5,6-b]-indoles have been prepared and tested for their antibacterial, antifungal, H1-antihistaminic and antimuscarinic activities. Among them, compounds 5b, 5d, 5k exhibited higher H1-antihistaminic activity than pheniramine maleate. Compounds 5c, 5d showed higher antibacterial activity than ampicillin against Staphylococcus aureus and E. coli, respectively. PMID- 12561238 TI - Synthesis of some new bis-thiazoles as possible anticancer agents. AB - Several new 5-(2,3-dihydrothiazol-2-yledinyl)rhodanines 3a-c and 5-(4 oxothiazolidinon-2-ylidenyl)rhodanine 4 were synthesized through the reaction of 5-thiocarbamoyl rhodanines 2 with phenacyl bromides or chloroacetic acid, respectively. The synthesis of the arylidene derivatives 5a-c were also described. The 5-(4-amino-5-cyano-2,3-dihydrothiazol-2-yledinyl)rhodanines 10a, b were obtained through reaction of rhodanines 1a, b with thiazolium salt 9. All the prepared compounds were screened for their anticancer activity using the NCI in vitro anticancer screening program. Three compounds showed promising anticancer activity against particular human cell lines used in the assay. PMID- 12561239 TI - [Benzo(f)furo(3,4-c)(2,7)naphthyridines from furo(3,4-b)pyridines]. AB - The pyridines 5, obtained by dehydrogenation of the 4-(2-nitrophenyl)-1,4 dihydropyridine lactones 4, give the tetracycles 6 and 7 by reduction of the nitro group with zinc/ammonium chloride followed by ring closure. Irradiation of the compounds 4 with UV-A light leads to the 4-(2-nitrosophenyl)pyridines 8. During the reaction of 8 with conc. hydrochloric acid the nitroso group cyclizes with the 5-acyl group yielding the chloro substituted tetracycles 9 and 10. The hydroxamic acids 6 and 9 are deoxygenated by phosphorus trichloride to yield the lactames 11, while O-acetylation with acetic anhydride affords 12. Phosphorus trichloride reduces the N-oxides 7 and 10 to the bases 13. Using the reaction with acetic anhydride the acetates 14 are formed by Boekelheide-transformation. Alcaline saponification of 14 gives the alcohols 15. PMID- 12561240 TI - [Unexpected formation of an estrone derivative from androsta-1,4-diene-3,17 dione]. AB - Reaction of androsta-1,4-diene-3,17-dione (1) with pyrrolidine and p toluenesulfonic acid in toluene gives 3-pyrrolidino-estra-1,3,5(10)-triene-6,17 dione (7) in poor yield, whose structure is shown by X-ray analysis. Compared with 3H-estradiol, compound 7 shows only a weak receptor binding activity tested in an in vitro screening using rabbit uterus cytosol. PMID- 12561241 TI - Validated HPLC method for determination of chlorzoxazone in human serum and its application in a clinical pharmacokinetic study. AB - A high performance liquid chromatographic (HPLC) method for the determination of chloroxazone in human serum using phenacetin as internal standard (IS) is described. Protein precipitation is used for preparation of the sample. A mobile phase consisting of acetonitrile and 0.5% acetic acid in water mixture (40:60 v/v) was used at a flow rate of 1 ml/min on a C18 column. The eluate was monitored using an UV/VIS detector set at 287 nm. Ratio of peak area of analyte to IS was used for quantification of serum samples. The absolute recovery was greater than 96% over a concentration range of 1 to 100 micrograms/ml and the limit of quantitation was 0.05 microgram/ml. The intra-day relative standard deviation (RSD) measured at 1, 10, 50, and 100 micrograms/ml ranged from 0.9 to 5.1%. The inter-day RSD ranged from 0.6 to 3.0%. The method is simple, sensitive and has been successfully used in pharmacokinetic study conducted in healthy human volunteers. PMID- 12561242 TI - A validated RP-HPLC method for the determination of mosapride citrate in bulk drug samples and pharmaceutical formulations. AB - Mosapride citrate, a selective serotonin 5-HT4 agonist, is a novel and potent gastroprokinetic drug. So far no assay procedure has been reported for the estimation of this drug either in bulk drug samples, pharmaceutical formulations or in biological samples. A rapid and sensitive high-performance liquid chromatographic method was developed for the estimation of mosapride citrate in bulk drug samples and pharmaceutical dosage forms. Risperidone was used as an internal standard (ISD). A HPLC system consisting of gradient pump, reverse phase C-18 analytical column, a variable UV-visible detector set at 274 nm and an integrator was used. The mobile phase consisted of acetonitrile: 0.02 M potassium dihydrogen phosphate buffer (pH adjusted to 4.0 with o-phosphoric acid) in the ratio of 50:50 (v/v), and was pumped at 1 ml/min at 40 degrees C. The drug and ISD were eluted at 8.10 and 2.27 min, respectively. The peak drug/ISD area ratio versus drug concentration relationship was linear (r = 0.9998). The method was validated for its linearity, precision and accuracy. The calibration curve was linear in the range of 0.5 to 30 micrograms/ml. The lower detection limit was found to be 0.23 microgram/ml. The intra- and inter-day variation was found to be less than 1% showing high precision of the assay method. The mean recovery of the drug from the solutions containing 2, 4 or 10 micrograms/ml was 101.55 +/- 0.97% indicating high accuracy of the proposed HPLC method. PMID- 12561243 TI - Rapid spectrophotometric method for quantitative determination of simvastatin and fluvastatin in human serum and pharmaceutical formulations. AB - A spectrophotometric method has been developed and applied to the determination of simvastatin and fluvastatin in human serum and in tablets. Simvastatine and fluvastatin were determined by measurement of their first derivative signals at 241.6, 245.9, 249.1 nm (for simvastatin) and 259.6 nm (for fluvastatin), respectively. Calibration curves were linear and the ranges of quantification were 12.0-28.0 micrograms.ml-1 for simvastatin and 10.0-28.0 micrograms.ml-1 for fluvastatin. The procedure was successfully applied to the determination of these compounds in pharmaceutical formulations as well as in human serum with a high percentage of recovery, good accuracy and precision, and without measurable interference by the excipients. PMID- 12561244 TI - Solubility, chemical and photochemical stability of curcumin in surfactant solutions. Studies of curcumin and curcuminoids, XXVIII. AB - The natural compound curcumin was incorporated into various micellar systems in order to improve the water solubility and the hydrolytic and photochemical stability. The presence of micellar structures resulted in an increase in water solubility at pH 5 by a factor of at least 10(5). The hydrolytic stability of curcumin under alkaline conditions was strongly improved by incorporation into micelles while the photodecomposition rate was increased compared to curcumin in hydrogen bonding organic solvents or aqueous solutions. The ability of curcumin to act as a photosensitizer was dependent on the type of micelles and pH of the medium. PMID- 12561245 TI - Differential scanning calorimetry to investigate the compatibility of ciprofloxacin hydrochloride with excipients. AB - The compatibility between ciprofloxacin hydrochloride (CFX) and some excipients was evaluated using differential scanning calorimetry (DSC). Physical mixture, coground mixture, compressed mixture and kneaded mixture were prepared to study the effect of sample manipulation. In addition, the samples of physical mixture were also accelerated at 55 degrees C for three weeks to obtain more reliable conclusions. Different types of excipients currently used in tablet or capsule formulations namely, calcium phosphate dibasic dihydrate (Emcompress), magnesium stearate lactose, sorbitol, mannitol, croscarmellose sodium (Ac-Di-Sol), sodium carboxymethyl starch (Primojel), microcrystalline cellulose (Avicel PH 101, Emcocil) were examined. The DSC scan of CFX displayed two endothermic peaks probably as a result of a fusion process followed by a decomposition process. CFX appeared to interact with sorbitol, mannitol, Ac-Di-Sol, Primojel, Avicel PH 101 and Emcocil. PMID- 12561246 TI - Metabolism of metamizol in early stages of the incubated hen's egg. AB - In a previous paper we introduced a model for studying the metabolism of xenobiotics in the incubated hen's egg. This model is characterized by application of the xenobiotic into the yolk sac and identification of metabolites in the allantoic fluid (AF). Depending on the stage of development, the incubated hen's egg can be used both conservatively--in the sense of an animal experiment- and alternatively as a complementary method (appr. first half of incubation period). In the present investigation we used the sodium salt of [(1,5-dimethyl-3 oxo-2-phenyl-2,3-dihydro-lH-pyrazol-4-yl)-N-methylamino] methanesulfonic acid (1; metamizol, dipyrone) as model substance. Concentrations of 1 up to 32 mg/egg did not affect the viability of the incubated eggs. After inoculation of 1 (10 mg/egg) on the sixth day of incubation (DI) eight compounds were identified in the AF on DI 11 (TLC, GC, GC-MS, synthesis, MS, 13C NMR): 4-methylamino-(2), 4 amino-(3), 4-hydroxy-(4), 4-acetylamino-(5), 4-N-acetyl-N-methylamino-(6), 4-N hydroxyacetyl-N-methylamino-(7), 4-N-hydroxyoxalyl-N-methylamino-(8) and 4-N formyl-N-methylamino-1,5-dimethyl-2-phenyl-1,2-dihydro-3H-pyrazol- 3-one ( antipyrine; 9), respectively. Main metabolites are 2 and 5; 4 and 7 occur as sulfate or glucuronide conjugates. Compound 7 is also eliminated in it's free form. To our knowledge 7 has been detected as a new metabolite of 1 for the first time. The metabolic route of 7 was elucidated after application of 6. Compounds 8 and 9 are artefacts of 7. The data presented here demonstrate the usefulness of the developed model also in early stages of the incubated hen's egg. Consequently, the model is suitable as a complementary method for xenobiotic metabolism. PMID- 12561247 TI - Physicochemical and microbiological properties as well as stability of ointments containing aloe extract (Aloe arborescens Mill.) or aloe extract associated to neomycin sulphate. AB - The aim of the study was to work out methods of quality assessment of ointments containing dry extract from fresh leaves of Aloe arborescens Mill. (Lilliaceae) and also of ointments containing both of dry extract and neomycin sulphate. The stability of the ointments, stored at 20 degrees C, was studied and the following criteria were considered: chromatographic analysis (TLC), pH of the ointments, the content of the substances in the dry extract converted to aloenin, the content of aloenin and aloin, anti-microbial activity of neomycin in the ointments, the size of the particles of the dry extract and of neomycin sulphate in the ointment suspension and the sterility of the ointments. After two years of storage at 20 degrees C, the ointments prepared with the anhydrous lipophilic base, did not change their physicochemical characteristics and neomycin in those ointments retained almost 100% of starting anti-microbial activity. Water or propylene glycol significantly decreased the stability of the biologically active substances of the dry extract in the ointments. Besides, in the ointments containing the dry extract and neomycin sulphate, the presence of water or propylene glycol induced degradation of the biologically active substances of the dry extract and a decrease in the anti-microbial activity of neomycin in the ointments. Considering the physicochemical and microbiological stability, the most advisable base for the ointments with aloe and neomycin sulphate was composed of white vaseline, liquid paraffin, solid paraffin, cholesterol. PMID- 12561248 TI - Transdermal delivery of glibenclamide and glipizide: in vitro permeation studies through mouse skin. AB - The purpose of this investigation was to study the feasibility of transdermal delivery of glibenclamide and glipizide. In vitro permeation of these drugs was studied through mouse skin using various penetration enhancers like Tween -20, polyethyleneglycol-400, ethanol and d-limonene by simultaneous application of drug and enhancer solution or by pretreatment of the skin with neat enhancer. The partition coefficient values indicated that both drugs partition well into the skin. Glipizide did not show any skin metabolism, while glibenclamide showed a minimal metabolism during in vitro skin metabolism studies. The flux values (microgram/cm2/h) of both drugs significantly (p < 0.05) increased in the presence of penetration enhancers. The glibenclamide flux values ranged from 1.39 +/- 0.13 without enhancer, to 19.01 +/- 2.14 in a combination of 50% ethanol and 5% d-limonene. Glipizide flux values ranged from 3.01 +/- 0.74 without enhancer, to 62.97 +/- 7.10 in a combination of 50% ethanol and 5% d-limonene. Skin retention and solubility of both drugs increased with all penetration enhancers compared to control. The target permeation rates for glibenclamide and glipizide were calculated to be 193.8 and 184.8 micrograms/h respectively. The present study showed that the target permeation rates for both drugs could be achieved with the aid of enhancers by increasing the area of application in an appreciable range. PMID- 12561249 TI - Effect of limonene on the in vitro permeation of nicardipine hydrochloride across the excised rat abdominal skin. AB - The aim of the present investigation was to study the effect of limonene on in vitro permeation of nicardipine hydrochloride across the excised rat abdominal skin from a 2% w/w hydroxypropyl cellulose (HPC) gel. The HPC gel formulations containing 1% w/w of nicardipine hydrochloride and selected concentrations of limonene (0% w/w to 12% w/w) were prepared, and subjected to in vitro permeation of the drug through excised rat abdominal epidermis. The drug content in the gels was found to be uniform and the drug was found to be stable in the HPC gel formulations. The permeation flux of nicardipine hydrochloride across rat epidermis was increased markedly by the addition of limonene to the HPC gels. A maximum flux was observed (246 +/- 1 micrograms/cm2/h) with an enhancement ratio of about 8 when limonene was incorporated at a concentration of 4% w/w. However, there was no further increase in the permeability of nicardipine hydrochloride beyond 4% w/w of limonene. The DSC and FT-IR data indicated that limonene increased the permeability of nicardipine hydrochloride across the rat epidermis by partial extraction of lipids in the stratum corneum. The results suggest that limonene may be useful for enhancing the skin permeability of nicardipine hydrochloride from transdermal therapeutic system containing HPC gel as a reservoir. PMID- 12561250 TI - Direct cytotoxicity of non-steroidal anti-inflammatory drugs in acidic media: model study on human erythrocytes with DIDS-inhibited anion exchanger. AB - Non-steroidal anti-inflammatory drugs (NSAID) elicit gastric damage through inhibition of the synthesis of prostaglandins that protect gastric cells and direct effect on mucous layer. As the latter effect is not well understood, we used acid hemolysis test in a model study on the cytotoxicity of nine NSAIDs. Human erythrocytes were used as model cells after their band 3 membrane protein was inhibited with DIDS (4,4'-diisothiocyano-2,2'-stilbenedisulfonate) that strongly suppressed the entry of acid into cytosole and postponed acid-induced hemolysis. These drugs did not produce measurable hemolysis in media buffered at pH 7.2. However, in acidic media (pH 3.4) they markedly reduced to a variable extent the prelytic interval (time spent by acid to accumulate overcritically in cytosole) and time for 50% hemolysis (acid resistance). The cytotoxicity of NSAID to erythrocytes at acidic medium was expressed by the inverse of the concentration (C50%) that reduced twofold acid resistance. It was related to the hydrophobicity of drug as the log of C50% depended linearly on the log of its critical concentration for the formation of micelles. Hence, the cytotoxicity of NSAIDs to model cells in acidic media apparently involved the transfer of protonated forms and accumulation of the drug and acid into cytosole. We conclude, the protonophore mechanism could be involved in the direct damage of erythrocytes in acidic media. Based on this cytotoxicity the NSAIDs were ranked as aspirin < paracetamol < nimesulide < diclofenac < piroxicam < meloxicam < ibuprofen < naproxen < indomethacin. This is roughly the same row that expresses the relative in vivo gastropathogenicity of NSAIDs, hence, it is likely this mechanism might damage gastric epithelial cells by generation of influx of NSAID and back diffusion of acid and producing stress conditions and apoptosis. PMID- 12561251 TI - Protection against damaged DNA in the single cell by polyphenols. AB - The protective properties of seven polyphenols against hydrogen peroxide induced DNA damage in human peripheral blood lymphocytes (PBL) were studied using single cell micro-gel electrophoresis. Hydrogen peroxide causes a concentration dependent increase in single cell DNA strand breakage in human PBL. Quercetin and 7,8-dihydroxy-4-methyl coumarin exhibited the strongest protection, significantly inhibiting 50 microM H2O2-induced DNA damage at a range of concentrations of 3.1 25 microM. Curcumin, resveratrol and vanillin protected against DNA damage induced by 50 microM H2O2 at a range of concentrations of 6.25-25 microM, but rutin and 7-hydroxy-4-methyl coumarin failed to provide any protection even at concentrations up to 50 microM. Quercetin, 7,8-dihydroxy-4-methyl coumarin, curcumin, resveratrol and vanillin are therefore effective in protection of human single cell DNA from oxidative attack. PMID- 12561252 TI - Antihepatotoxic activity of Phyllanthus fraternus. AB - Different fractions of alcoholic extracts of aerial parts and roots of Phyllanthus fraternus Webster (Euphorbiaceae) were screened for antihepatotoxic activity on carbon tetrachloride induced liver damage in albino rats. The degree of protection was measured using biochemical parameters like serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvate transaminase (SGPT), total protein (TP) and total albumin (TA). The methanol fraction was found to be the most active, which was further supported by a significant recovery of hepatocytes in the histopathological study of the liver showing almost complete normalization of the tissues as neither the fatty accumulation nor the necrosis was observed. PMID- 12561253 TI - Two flavones from Scutellaria baicalensis Georgi and their binding affinities to the benzodiazepine site of the GABAA receptor complex. AB - A new flavone 6,2'-dihydroxy-5,7,8,6'-tetramethoxyflavone (1) together with one known flavone 5,7,2'-trihydroxy-6,8-dimethoxyflavone (2) were isolated from the roots of Scutellaria baicalensis Georgi. Their structures were elucidated on the basis of spectral evidence and their affinities for the benzodiazepine (BDZ) site of the GABAA receptor complex were evaluated with a radioligand receptor binding assay. PMID- 12561254 TI - Acylated flavonol glycoside from Psidium gauijava L. seeds. AB - Ten phenolic and flavonoid compounds including one new acylated flavonol glycoside were isolated from Psidium gauijava seeds. The structures of the new compound quercetin-3-O-beta-D-(2"-O-galloyl glucoside)-4'-O-vinylpropionate and of the known compounds were elucidated by different chemical and physical methods, 1H- and 13C NMR spectral analysis. PMID- 12561255 TI - Describing solubility of polymorphs in mixed solvents by CNIBS/R-K equation. PMID- 12561256 TI - Oral antiplatelet therapies have no effect on circulating levels of RANTES in patients with coronary artery disease. PMID- 12561257 TI - Jacobson and Soliman's analysis of the public health effects of tobacco and firearm litigation. PMID- 12561258 TI - Thoughts in dark times of a world made new. PMID- 12561259 TI - Introduction: merging law, human rights, and social epidemiology. PMID- 12561260 TI - Integrating law and social epidemiology. PMID- 12561261 TI - Health and human rights: old wine in new bottles? PMID- 12561262 TI - HIV and the law: integrating law, policy, and social epidemiology. PMID- 12561263 TI - Criminal law, policing policy, and HIV risk in female street sex workers and injection drug users. PMID- 12561264 TI - Does indebtedness influence health? A preliminary inquiry. PMID- 12561265 TI - Zoning law, health, and environmental justice: what's the connection? PMID- 12561266 TI - Thinking about AIDS and stigma: a psychologist's perspective. PMID- 12561267 TI - The impact of law on coronary heart disease: some preliminary observations on the relationship of law to "normalized" conditions. PMID- 12561268 TI - Patents on drugs: manufacturing scarcity or advancing health? PMID- 12561269 TI - Health and the governance of security: a tale of two systems. PMID- 12561270 TI - Juridical action for the protection of collective rights and its legal impact: a case study. PMID- 12561271 TI - Rethinking health and human rights: time for a paradigm shift. PMID- 12561272 TI - Regulation of the global marketplace for the sake of health. PMID- 12561273 TI - Human rights and public health: dichotomies or synergies in developing countries? Examining the case of HIV in South Africa. PMID- 12561274 TI - How the law affects gun policy in the United States: law as intervention or obstacle to prevention. PMID- 12561275 TI - Co-opting the health and human rights movement. PMID- 12561276 TI - Review of methodologies measuring human rights implementation. PMID- 12561277 TI - Human rights and wrongs: could health impact assessment help? PMID- 12561278 TI - The evolving field of health and human rights: issues and methods. PMID- 12561279 TI - Tobacco litigation: medical monitoring of healthy smokers. PMID- 12561280 TI - Duty to warn: learned intermediary doctrine not applicable when pharmacist knows of drug contraindication. PMID- 12561281 TI - Administrative law: rulemaking authority and standing to sue for office surgery procedures. PMID- 12561283 TI - A review of 50 years of research on naturally occurring family routines and rituals: cause for celebration? AB - This article is a qualitative review of 32 publications appearing since J. Bossard and E. Boll's (1950) seminal work on family rituals was conducted. Definitions are offered whereby a distinction is made between family routines as observable practices and family rituals as symbolic representations of collective events. The relative occurrence of family routines, as described in the literature, appears to follow a developmental course and is affected by the cultural environment. Family routines and rituals were found to be related to parenting competence, child adjustment, and marital satisfaction. The studies were limited by inconsistent methods of assessing family routines, reliance on samples of convenience, and a failure to distinguish between direct and indirect effects. Recommendations are made to better integrate theory with empirical efforts to demonstrate the importance of family routines and rituals in contemporary life. PMID- 12561284 TI - Intimate culture of families in the early socialization of literacy. AB - The intimate family culture for early literacy socialization was documented for a socioculturally heterogeneous sample of 66 children enrolled in pre-kindergarten through third grade at public elementary schools in a large U.S. city. Parents were interviewed about 3 types of indexes of their family's intimate culture: the child's engagement in various literacy-related activities at home, the parents' orientation towards the significance of literacy for early child development, and the family's routines of dinnertime, reading aloud, and doing homework for school. Basic reading competencies were assessed with the Woodcock-Johnson Psychoeducational Battery--Tests of Achievement, Revised (1989). Multiple regression analysis found that a significant proportion of variance in the children's literacy development was predicted by each of the quantitative indexes of intimate family culture, leaving little or no additional variance that was due to family income or ethnicity. PMID- 12561285 TI - Adolescent satisfaction in family rituals and psychosocial development: a developmental systems theory perspective. AB - Adolescent satisfaction in family rituals and psychosocial development (E. Greenberger & A. B. Sorenson, 1974) were explored in the context of adolescent personality characteristics (International Personality Item Pool, 1999) and family environment characteristics (S. M. Gavazzi, M. J. Reese, & R. M. Sabatelli, 1998; D. H. Olson et al., 1983). Data were collected from 159 female undergraduates with the Adolescent Satisfaction in Family Rituals Scale (D. G. Eaker & L. H. Walters, 1999). Family ritual satisfaction was positively related to late adolescent psychosocial development and mediated the relation between family boundaries and psychosocial development. Furthermore, the relation between personality (measured as discontentedness, an aspect of neuroticism) and satisfaction with family rituals was found to be mediated by family boundaries in preliminary analyses. These results suggest that the relevance of family ritual experiences to adolescent psychosocial development is in part a function of an individual's personality and the family environment. PMID- 12561286 TI - Longitudinal changes in the time parents spend in activities with their adolescent children as a function of child age, pubertal status, and gender. AB - This study examined the time Dutch mothers (N = 301) and fathers (N = 255) spend per day engaging in 4 activities (going somewhere, doing something, watching TV, and eating together) with their adolescent children both concurrently and 5 years later. Also assessed was whether parent-child shared time was related to parent or child gender and whether age-related differences could be explained by adolescent pubertal status, family conflict, adolescent and parent work or volunteer hours, parental work stress, and adolescent computer use. Finally, the study examined whether family conflict predicted changes in shared time and whether shared time predicted changes in conflict. The findings showed that age changes depended on the activity and that pubertal status mediated age differences in TV viewing among mixed-gender parent-child pairs. Shared time during pre-, early, and mid-adolescence was linked to decreases in family conflict 5 years later. PMID- 12561287 TI - Inclusion and belonging in the family rituals of gay and lesbian people. AB - Survey data collected from 400 non-metropolitan gay men and lesbians were used to examine what factors lead them to attend a family-of-origin ritual and affect their sense of belonging during the event. The present study was inspired by qualitative findings regarding the production of outsider status during rituals. Attendance and belonging were both predicted by type of ritual and the quality of relationships with families of origin. Also, partners were more likely to be invited when the couple relationship was more visible. Residential community climate, age, income, and gender were not significant. The family membership complexities of gay and lesbian people are discussed, and a more nuanced understanding of membership during ritual is encouraged. PMID- 12561288 TI - Integrating family routines and rituals with other family research paradigms: comment on the special section. AB - The articles in this special section represent cutting edge research on family routines and rituals. This comment discusses how this work may inform and be informed by other family research. This includes studying how routines develop, are maintained, and reorganize as well as how they influence individual outcomes through effects on socialization, security, and adaptation to stress. Applications of research on family routines to enhance prevention and treatment are also discussed. New directions for the study of family rituals are discussed, including how ritual events may have long-term effects on family interaction through discontinuous changes in relationship expectations of family members. PMID- 12561289 TI - Family routines and rituals--the importance of family functioning: comment on the special section. AB - This diverse collection of papers highlights routines and rituals in family life. Just as individual family members bring unique styles and perspectives to the family table, to be incorporated into a family framework, so do each of these papers present rich and varied research questions, designs, and measurement strategies that enrich our understanding of family routine behavior patterns and ritual meanings. This series adds to our conceptualization of family routines and rituals within a systems perspective highlighting: 1) families are comprised of multiple levels that operate individually; interact with each other; and as a whole, reveal properties distinct from separate parts; 2) families tend toward stability in meaningful patterns of functioning; and 3) family functioning has meaning for individual outcomes. PMID- 12561290 TI - Family rituals--from research to the consulting room and back again: comment on the special section. AB - This article offers a comment on the special research section on Family Routines and Rituals from the point of view of a family therapist who has made the use of rituals a specialty in clinical practice. Ways that the research usefully informs clinicians seeking to use rituals in therapy are highlighted. A critique of the articles, particularly in the area of ritual definition, is offered. The comment concludes with several suggestions for future research. PMID- 12561291 TI - Parents' emotional availability and infant emotional competence: predictors of parent-infant attachment and emerging self-regulation. AB - One-year-old infants (N = 62) and their mothers and fathers were observed in free play and teaching sessions in order to examine parents' emotional availability and the infant's emotional competence. Mothers were more emotionally available than fathers, and infants exhibited more effortful attention with mothers than with fathers. Similar relations between parental emotional availability and infant emotional competence were found for mother-infant and father-infant dyads. Change in parental emotional availability covaried with change in infant emotional competence. Individual differences in parental emotional availability and infant emotional competence were more consistent across contexts than across parents. Infant effortful attention at 12 months was a mediator between maternal emotional availability at 12 months and toddler situational compliance at 16 months. PMID- 12561292 TI - Marital conflict and boys' peer relationships: the mediating role of mother-son emotional reciprocity. AB - This study examined the role of mother-son emotional reciprocity in connections between marital conflict and the quality of boys' peer relationships. Parents from 84 intact families with preadolescent boys reported on the level of conflict in their marital relationship. Observations of mother-son interaction were coded for emotional reciprocity, and assessments of boys' peer relationships were obtained from both teachers and classmates. No direct connection between marital conflict and boys' peer relationships was observed. Rather, marital conflict affected boys' social competence indirectly, through its effect on the emotions expressed between mother and son. The findings support the proposal that emotional processes play an important role in connections between marital conflict and children's peer relationships and suggest that family emotional expressiveness deserves greater attention in both research and intervention. PMID- 12561293 TI - Children's responses to mothers' and fathers' emotionality and tactics in marital conflict in the home. AB - Addressing a gap in methodological approaches to the study of links between marital conflict and children, 51 couples were trained to complete home diary reports on everyday marital conflicts and children's responses. Parental negative emotionality and destructive conflict tactics related to children's insecure emotional and behavioral responses. Parental positive emotionality and constructive conflict tactics were linked with children's secure emotional responding. When parents' emotions and tactics were considered in the same model, negative emotionality was more consistently related to children's negative reactions than were destructive conflict tactics, whereas constructive conflict tactics were more consistently related to children's positive reactions than parents' positive emotionality. Differences in children's responding as a function of specific parental negative emotions (anger, sadness, fear) and parent gender were identified. PMID- 12561294 TI - Distributions of caregiving tasks among family members: the place of gender and availability. AB - To extend family-oriented approaches to caregiving, participants in 2 studies were asked to distribute tasks among a set of adult children, first with information only about gender and then with systematically varied information about commitments to paid work, marriage, and/or parenting. Making the distributions, using a computer-based program, were 2 groups of older adults (ages 60 to 90 years). In Study 1, gender composition was kept constant (2 sons and 2 daughters). In Study 2, it was varied. The results showed several ways in which people combine attention to gender and to availability. The results also pointed to the need to consider both the number and type of tasks allocated. The results are discussed in terms of implications for the way caregiving is regarded, the development of multiple-factor models for variations among family members, and the possible replications and extensions to other circumstances and populations. PMID- 12561295 TI - Perceiving the division of family work to be unfair: do social comparisons, enjoyment, and competence matter? AB - Past research has revealed that women, working or not, perform more family labor (i.e., housework and child care) than do men. Yet, women often do not perceive this as unfair. Drawing on the theoretical work of L. Thompson (1991) and B. Major (1993), the authors hypothesized that this paradox might be explained by women perceiving greater fairness in the lopsided division of family work (a) when they compare the amount of family work they do with other women (who perform similar amounts) rather than with the spouse; (b) when they enjoy performing family work; and/or (c) when they and their spouses believe that they are especially competent at family work. Data from a 3-panel, longitudinal study of married couples expecting their first child were consistent with the second and third predictions but not the first. Further, prospective analyses suggested that perceiving inequity in family work leads wives and husbands to make more frequent comparisons with the spouse and sometimes with same-sex others. PMID- 12561296 TI - ORYX data to play key role in new JCAHO survey process. PMID- 12561297 TI - Tabletop drills not enough for testing disaster plan. PMID- 12561298 TI - VA's surgical QI program could be available to all. PMID- 12561299 TI - Improve performance by taking outsiders' view. Part 2. PMID- 12561300 TI - HHS launches national nursing home QI. PMID- 12561301 TI - Patient safety alert. Construction brings opportunity to boost patient safety. PMID- 12561302 TI - Test interventions for improving safety. PMID- 12561303 TI - 'Everybody wins' as CM, home health join forces. PMID- 12561304 TI - A quantum-chemical study of dinitrogen reduction at mononuclear iron-sulfur complexes with hints to the mechanism of nitrogenase. AB - The mechanism of biological dinitrogen reduction is still unsolved, and the structure of the biological reaction center, the FeMo cofactor with its seven iron atoms bridged by sulfur atoms, is too complicated for direct attack by current sophisticated quantum chemical methods. Therefore, iron-sulfur complexes with biologically compatible ligands are utilized as models for studying particular features of the reduction process: coordination energetics, thermodynamic stability of intermediates, relative stability of isomers of N2H2, end-on versus side-on binding of N2, and the role of states of different multiplicity at a single iron center. From the thermodynamical point of view, the crucial steps are dinitrogen binding and reduction to diazene, while especially the reduction of hydrazine to ammonia is not affected by the transition metal complex, because the complex-free reduction reaction is equally favored. Moreover, the abstraction of coordinated ammonia can be easily achieved and the complex is recovered for the next reduction cycle. Our results are discussed in the light of studies on various model systems in order to identify common features and to arrive at conclusions which are of importance for the biological mechanism. PMID- 12561305 TI - An intramolecular N-H...(mu-H)Re2 dihydrogen bond and a novel mu 3-eta 2 coordination mode of the pyrazolate anion on a triangular cluster face. AB - The quantitative addition of pyrazole (Hpz) to the 44 valence-electron, triangular cluster anion [Re3(mu 3-H)-(mu-H)3(CO)9]- gives the novel unsaturated anion [Re3(mu-H)4(CO)9(Hpz)]- (1, 46 valence electrons), which contains a pyrazole molecule that is terminally coordinated on a cluster vertex. Solidstate X-ray and IR analyses reveal a rather weak hydrogen-bonding interaction between the NH proton and one of the hydrides bridging the opposite triangular cluster edge (delta H degree = -3.1 kcal mol-1 from the Iogansen equation). Both IR and NMR data indicate that such a proton-hydride interaction is maintained in the major conformer present in CD2Cl2, but also provide evidence of the presence of minor conformers of 1 in which the NH proton is involved in an intermolecular hydrogen bond with the solvent. The mu-H...HN bond length evaluated in solution through the T1 minimum value (2.07 A) and that determined in the solid state by X ray diffraction (2.05 A) are in good agreement. NMR experiments show that, in acetone, intermolecular N-H...solvent interactions replace the intramolecular dihydrogen bond. At room temperature in CH2Cl2, the pyrazole ligand in 1 is labile and 1 slowly "disproportionates" to [Re3(mu 3-H)-(mu-H)3(CO)9]- and [Re3(mu-H)3(CO)9-(mu-eta 2-pz)(Hpz)]-, with H2 evolution. Slow H2 evolution also leads to the formation of the anion [Re3(mu-H)3-(CO)9(pz)]- (5), in which the pyrazolate anion adopts a novel mu 3-eta 2-coordination mode, as revealed by a single-crystal X-ray analysis. The analysis of the bond lengths indicates that the pyrazolate anion in 5 acts as a six-electron donor, with loss of the aromaticity. The formation of 5 from 1 is much faster in solvents with a high dielectric constant, such as acetone or DMF. Anion 5 was also obtained from the reaction of pyrazole with [Re3(mu-H)3(CO)9(mu 3-CH3)]- through the intermediate formation of two isomeric addition derivatives and following CH4 evolution. PMID- 12561306 TI - Mechanisms for NH3 decomposition on Si(100)-(2 x 1) surface: a quantum chemical cluster model study. AB - In this paper, we present a detailed mechanism for the complete decomposition of NH3 to NHx(a) (x = 0-2). Our calculations show that the initial decomposition of NH3 to NH2(a) and H(a) is facile, with a transition-state energy 7.4 kcal mol-1 below the vacuum level. Further decomposition to N(a) or recombination-desorption to NH3(g) is hindered by a large barrier of approximately 46 kcal mol-1. There are two plausible NH2 decomposition pathways: 1) NH2(a) insertion into the surface Si-Si dimer bond, and 2) NH2(a) insertion into the Si-Si backbond. We find that pathway (1) leads to the formation of a surface Si = N unit, similar to a terminal Si = Nt pair in silicon nitride, Si3N4, while pathway (2) leads to the formation of a near-planar, subsurface Si3N unit, in analogy to a central nitrogen atom (Nc) bounded to three silicon atoms in the Si3N4 environment. Based on these results, a plausible microscopic mechanism for the nitridation of the Si(100)-(2 x 1) surface by NH3 is proposed. PMID- 12561307 TI - Use of low-temperature thermal alkylation to eliminate ink migration in microcontact printed patterns. AB - We demonstrate aqueous hydrogel-based microcontact printing of amine ligands into solvent-templated nanocavities of chloromethylphenyl-based siloxane or thin polymer films. Migration of pyridine ligands within films following printing, which can compromise pattern fidelity, is eliminated by heat treatment of the substrate. Gentle heating (e.g., 50 degrees C, 5 min) leads to the efficient alkylation of mobile pyridine adsorbate by the C-Cl bonds of the film, covalently tethering the adsorbate to the surface as a pyridinium salt. Subsequent binding of a Pd-based colloid to surface pyridinium (and remaining strongly bound and immobile pyridine ligand) sites permits selective electroless metal deposition and fabrication of patterned metal films. PMID- 12561308 TI - Covalently bonded platinum(II) complexes of alpha-amino acids and peptides as a potential tool for protein labeling. AB - Arylplatinum(II) complexes have been covalently bonded to the N and C termini and to the alpha-carbon of various amino acid derivatives. These organometallic functionalized amino acid compounds can be converted into the corresponding free amino acids under both basic and acidic conditions; this demonstrates the excellent stability properties of these biomolecules. Due to the NMR activity displayed by the 195Pt nucleus (natural abundance 33.8%, I = 1/2) these compounds are functional bio-markers. Furthermore, the ability of the arylplatinum functional group to bind SO2 gas, selectively and reversibly as indicated by changes in the spectroscopic properties (1H, 13C, 195Pt NMR and UV spectra) of these compounds, allows for the potential use of these complexes as in vitro biosensors. PMID- 12561309 TI - A novel and useful oxidative intramolecular coupling reaction of phenol ether derivatives on treatment with a combination of hypervalent iodine(III) reagent and heteropoly acid. AB - The oxidative intramolecular coupling reaction of phenol ether derivatives (nonphenolic derivatives) on treatment with a novel combination of a hypervalent iodine(III) reagent, phenyliodine bis(trifluoroacetate) (PIFA), and heteropoly acid (HPA) was studied. Biaryl compounds were obtained in excellent yields on treatment of highly substituted phenol ethers. On the other hand, spirodienones were specifically formed when one of the preferred arylic coupling sites was substituted with a methoxy group in the para position. PMID- 12561310 TI - Oligomerization and hydroamination of terminal alkynes promoted by the cationic organoactinide compound [(Et2N)3U][BPh4]. AB - The three ancillary amido moieties in the cationic complex [(Et2N)3U][BPh4] are highly reactive and are easily replaced when the complex is treated with primary amines. The reaction of [(Et2N)3U][BPh4] with excess tBuNH2 allows the formation of the cationic complex [(tBuNH2)3(tBuNH)3U][BPh4]. X-ray diffraction studies on the complex indicate that three amido and three amine ligands are arranged around the cationic metal center in a slightly distorted octahedral mer geometry. The cationic complex reacts with primary alkynes in the presence of external primary amines to primarily afford the unexpected cis dimer and, in some cases, the hydroamination products are obtained concomitantly. The formation of the cis dimer is the result of an envelope isomerization through a metal-cyclopropyl cationic complex. In the reaction of the bulkier alkyne tBuC identical to CH with the cationic uranium complex in the presence of various primary amines, the cis dimer, one trimer, and one tetramer are obtained regioselectively, as confirmed by deuterium labeling experiments. The trimer and the tetramer correspond to consecutive insertions of an alkyne molecule into the vinylic CH bond trans to the bulky tert-butyl group. The reaction of (TMS) C identical to CH with the uranium catalyst in the presence of EtNH2 followed a different course and produced the gem dimer along with the hydroamination imine as the major product. However, when other bulkier amines were used (iPrNH2 or tBuNH2) both hydroamination isomeric imines Z and E were obtained. During the catalytic reaction, the E (kinetic) isomer is transformed into the most stable Z (thermodynamic) isomer. The unique reactivity of the alkyne (TMS) C identical to CH with the secondary amine Et2NH is remarkable because it afforded the trans dimer and the corresponding hydroamination enamine. The latter probably results from the insertion of the alkyne into a secondary metal-amide bond, followed by protonolysis. PMID- 12561311 TI - Adaptation of the Rothemund reaction for carbaporphyrin synthesis: preparation of meso-tetraphenylazuliporphyrin and related benzocarbaporphyrins. AB - Electrophilic substitution of azulene has recently been shown to provide the means by which carbon-carbon bonds can be generated to form novel macrocyclic systems such as calixazulenes. These studies inspired us to develop a "one-pot" Rothemund-type synthesis of meso-tetraphenylazuliporphyrin. Azuliporphyrins, a group of cross-conjugated carbaporphyrinoids that exhibit intriguing chemistry and metallation properties, have previously only been available by multistep syntheses. In this work, azulene, pyrrole and benzaldehyde were shown to react in a 1:3:4 ratio in the presence of boron trifluoride etherate to give meso tetraphenylazuliporphyrin 7a. The free base shows only a minor diatropic ring current, but addition of TFA generates the related dication which shows greatly enhanced diatropicity where the internal CH shifts from delta = +3.35 to -0.5 ppm. Addition of pyrrolidine to 7a gave rise to a carbaporphyrin adduct which showed a porphyrin-like UV/Vis spectrum and the internal CH shifted further upfield to give a resonance near delta = -5.7 ppm. Treatment of 7a with tertbutyl hydroperoxide in the presence of potassium hydroxide afforded a mixture of benzocarbaporphyrins 9a-c. These tetraphenylcarbaporphyrins were fully aromatic by NMR spectroscopy and gave typical porphyrin-type UV/Vis spectra with a strong Soret band near 446 nm. This new methodology makes these important porphyrin analogues readily available for further study. PMID- 12561312 TI - A direct link between annulene and porphyrin chemistry--21-vacataporphyrin. AB - A novel molecule, aza-deficient porphyrin 5,10,15,20-tetraaryl-21-vacataporphyrin has been synthesised by a substraction of a tellurium atom from 5,10,15,20 tetraaryl-21-telluraporphyrin under treatment of HCl. The new macrocycle is an annulene-porphyrin hybrid and at the same time is directly related to 21 heteroporphyrins but has a vacant space instead of heteroatomic bridge. The molecule preserves the fundamental structural and spectroscopic features of the parental 5,10,15,20-tetraarylporphyrin with three nitrogen atoms and two CH groups favorably prearranged for coordination. PMID- 12561313 TI - Photoinduced electron transfer between metal-coordinated cyclodextrin assemblies and viologens. AB - Two novel tris(bipyridine)ruthenium(II) complexes bearing two and six beta cyclodextrin binding sites on their ligands have been synthesised and characterised. Complex 1, bearing two cyclodextrins, adopts a conformation in aqueous solution where parts of the aromatic ligands are self-included into the cyclodextrin moieties. This results in a loss of symmetry of the complex and gives rise to a much more complicated 1H NMR spectrum than expected. Photophysical studies indicate that the appended cyclodextrins protect the luminescent ruthenium core from quenching by oxygen, which results in longer excited state lifetimes and higher emission quantum yields compared with the reference compound, the unsubstituted ruthenium tris(bipyridine). Inclusion of suitable guests such as dialkyl-viologens leads to a quenching of the luminescence of the central unit. In these supramolecular donor-acceptor dyads an efficient photoinduced electron transfer from the excited ruthenium moiety (the donor) to the viologen unit (the acceptor) is observed. The alkyl chain length of the acceptor plays an important role on the binding properties; when it exceeds a certain limit the binding becomes strong enough for electron transfer to occur. Interestingly, a viologen with only one long alkyl tail instead of two shows no efficient quenching; this indicates that cooperative interactions between two cyclodextrins binding one viologen are essential to raise the binding constant of the supramolecular dyad. PMID- 12561314 TI - Orientational effect on the photophysical properties of quaterthiophene-C60 dyads. AB - Two quaterthiophene-[60]fullerene dyads in which C60 is singly (4TsC) or doubly (4TdC) connected to the inner beta-position of the terminal thiophene rings have been synthesized. The electronic properties of these donor-acceptor compounds were analyzed by UV/Vis spectroscopy and cyclic voltammetry, and their photophysical properties in solution and in the solid state by (time-resolved) photoluminescence (PL) and photoinduced absorption (PIA) spectroscopy. Both the flexible and geometrically constrained 4TsC and 4TdC dyads exhibit photoinduced charge transfer from the quaterthiophene to the fullerene in toluene and o dichlorobenzene (ODCB). In toluene, charge transfer occurs in both dyads by an indirect mechanism, the first step of which is a singlet-energy transfer from the 4T(S1) state to the C60(S1) state. In the more polar ODCB, direct electron transfer from 4T(S1) competes with energy transfer, and both direct and indirect charge transfers are observed. The geometrical fixation of the donor and acceptor chromophores in 4TdC results in rate constants for energy and electron transfer that are more than an order of magnitude larger than those of the flexible 4TsC system. For both dyads, charge recombination is extremely fast, as inferred from picosecond-resolved temporal evolution of the excited state absorption of the 4T.+ radical cation both in toluene and ODCB. PMID- 12561315 TI - Dinuclear CoII/GdIII and CoIII/GdIII complexes derived from hexadentate Schiff bases: synthesis, structure, and magnetic properties. AB - Two heterodimetallic complexes of formulae [LCo(MeOH)Gd (NO3)3] (1) and [LCo(AcO)2Gd(NO3)2] (2) (H2L = 1,3-bis[(3-methoxysalicylidene)amino]-2,2' dimethylpropane) have been synthesized and characterized. The structure of 1 consists of discrete dinuclear entities. The cobalt(II) ion exhibits a square pyramidal geometry, in which the basal plane is formed by the N2O2 set of the inner Schiff base site and the apical position is occupied by the methanol oxygen atom. The gadolinium(III) ion is ten-coordinate to three bidentate nitrate groups and the four oxygen atoms of the Schiff base. The phenolate oxygen atoms act as a bridge between both metal ions. Complex 2 is also formed by isolated dinuclear species. The cobalt(III) ion shows a distorted octahedral geometry in which the equatorial plane is formed by the N2O2 set of the Schiff base, and the axial positions are occupied by two oxygen atoms from both acetate groups. The gadolinium(III) ion is ten-coordinate to two bidentate nitrate groups, two oxygen atoms of the acetate groups, and the four oxygen atoms of the Schiff base. The metal ions are bridged through both the phenolate oxygen and the acetate groups, the latter acting as mu 2 ligands. Magnetic measurements on compound 1 allowed, for the first time, a quantitative evaluation of the J(Co,Gd) ferromagnetic interaction parameter (J = 0.90 cm-1). The CoII zero-field splitting has to be taken into account to fit the experimental data at low temperature (D = 4.2 cm 1). In complex 2, the magnetically isolated gadolinium center obeys a Curie law. PMID- 12561316 TI - Chemical triple-mutant boxes for quantifying cooperativity in intermolecular interactions. AB - Chemical double-mutant cycles have been used to quantify intermolecular functional-group interactions in H-bonded zipper complexes in chloroform. If the same interaction is measured in zippers of different overall stability, the double-mutant cycles can be combined to produce a triple-mutant box. This construct quantifies cooperativity between the functional group interaction of interest and the other interactions that are used to change the overall stability of the complexes. The sum of two edge-to-face aromatic interactions (-2.9 +/- 0.5 kJ mol-1) is shown to be insensitive to changes of up to 13.7 +/- 0.2 kJ mol-1 in the overall stability of the complex. In principle, enthalpic cooperative effects caused by entropy-enthalpy compensation could perturb the measurement of intermolecular interactions when using the double-mutant cycle approach, but these experiments show that, for this system, the magnitude of the effect lies within the error of the measurements. PMID- 12561317 TI - Chemoenzymatic syntheses of linear and branched hemithiomaltodextrins as potential inhibitors for starch-debranching enzymes. AB - Oligosaccharides embodying the S-maltosyl-6-thiomaltosyl structure have been readily synthesised by using convergent chemoenzymatic approaches. The key steps for the preparation of these molecules involved: 1) transglycosylation reactions of maltosyl fluorides onto suitable acceptors catalysed by the bacterial transglycosylase, cyclodextrin glycosyltransferase (CGTase), and 2) the SN2-type displacement of a 6-halide from acetylated acceptors by activated 1-thioglycoses. The target molecules, which were obtained in good overall yields, proved to be useful for investigating substrate binding in the active sites of several enzymes that act upon the alpha-1,6-linkage of pullulan and/or amylopectin. The compounds exhibit Ki values in the 2.5-1350 microM range with the different enzymes, and the highest affinity found by using these molecules was seen for the pullulanase from Bacillus acidopullulyticus. Both barley-malt limit dextrinase and pullulanase type II from Thermococcus hydrothermalis only recognised the longest linear thiooligosaccharide, while a branched heptasaccharide was the strongest inhibitor of pullulanase from Klebsiella planticola. PMID- 12561318 TI - Dimorphic intra- and intermolecular aryl motifs in symmetrical hexafaceted molecules (ArnX)3Y-Z-Y(XArn)3. AB - A variety of crystals containing molecules formulated generally as (ArnX)3Y-Z Y(XArn)3 with exact or pseudo S6 symmetry have been analysed for the presence of intra- and intermolecular multiple aryl embraces composed of offset-face-to-face (OFF) and edge-to-face (EF) local motifs. Ar is phenyl or substituted phenyl; n = 1, 2 or 3; the linkage Z is linear diatomic, linear monatomic, absent (Y-Y bond), or void (Y atoms unconnected). Representative molecular systems analysed are (PhCH2)3Si-O-Si(CH2Ph)3, (PhCH2)3Sn-Sn(CH2Ph)3, (4-Me-C6H4CH2)3Ge-O-Ge(CH2C6H4-4 Me)3, [(PhS)3Fe-O-Fe(SPh)3]-, [(PhCH2)3P-Cu-P(CH2Ph)3]+ and [(Ph3P)3Ag-CN Ag(PPh3)3]+. One characteristic intramolecular motif is a sixfold phenyl embrace (6PE) in which aryl groups fold back to the central domain of the molecule forming a rhombohedral box maintained by (EF)6. When the aryl groups are in exo positions there is an (EF)3 motif, like a tractor wheel, at each end of the molecule: [(Ph2N)3Ti-O-Ti(NPh2)3] possesses both intramolecular motifs as (EF)3 (EF)6-(EF)3. The molecules in this set are hexafaceted, and the crystal packing is generally based on intermolecular EF or OFF motifs with these faces, either from the central (EF)6 set or the end (EF)3-(EF)3 sets. Three types of subtle crystal packing isomerism occur: 1) the faces of the rhombohedral boxlike molecules slide over each other with variation of the intermolecular motifs, forming dimorphic crystals in space groups R3 or P1; 2) the faces of the tractor wheels (EF)3 slide over each other or 3) very similar molecules crystallise with the rhombohedral box or tractor wheel structures. In general the molecules considered are shape auspicious rather than shape awkward; solvent is included in a small proportion of the crystals and the crystal packing in these compounds is evidently efficient. Some principles for elaboration of these systems and the design of molecular crystal lattices are considered. PMID- 12561319 TI - Self-organized lipid-porphyrin bilayer membranes in vesicular form: nanostructure, photophysical properties, and dioxygen coordination. AB - An amphiphilic tetraphenylporphyrin and its iron complex bearing four phospholipid substituents, in which a trimethylolethane residue connects the two acyl chains (lipid-porphyrins), have been synthesized. The free-base lipid porphyrin 6a self-organizes in aqueous media to form spherical unilamellar vesicles with a diameter of 100 nm and a uniform thickness of 10 nm, which corresponds to twice the length of the molecule. In the visible absorption spectrum, the porphyrin Soret band was significantly red-shifted (12 nm) relative to that of the monomer in benzene/MeOH solution due to the excitonic interaction of the porphyrin chromophores. The [symbol: see text]-A isotherm of 6a gave an area per molecule of 2.2 nm2, which allowed the estimation of the number of molecules in a single vesicle (2.3 x 10(4)). Double-layered Langmuir-Blodgett (LB) films of 6a on a glass surface exhibited an absorption spectrum identical to that of the 6a vesicles in bulk aqueous solution, and this suggests that they contain similar geometric arrangements of the porphyrin moieties. Exciton calculations on the basis of our structural model reproduced the bathochromic shift of the Soret band well. In the photophysical properties of the 6a vesicles, the characteristics of J-aggregated porphyrins substantially predominate: strong fluorescence and extremely short triplet lifetime. The iron complex 6b with a small molar excess of 1-dodecylimidazole (DIm) also formed spherical unilamellar vesicles (100 nm phi). Scanning force microscopy after evaporation on a graphite surface revealed 6b/DIm vesicles with a vertical height of 19.8 nm, which coincided with the thickness of the double bilayer membranes. The ferrous 6c formed a bis(DIm)-coordinated low-spin FeII complex under an N2 atmosphere. Upon addition of O2 to this solution, a kinetically stable O2 adduct was formed at 37 degrees C with a half-life of 17 h. Distinct gel-phase (liquid-crystal) transitions of the lipid-porphyrin membranes were clearly observed; the free base 6a displayed a higher transition temperature (56 degrees C) than the iron complex. Magnetic circular dichroism and infrared spectroscopic studies proved that molecular O2 coordinates to the self-organized lipid-porphyrinatoiron(II) vesicles in aqueous media. PMID- 12561320 TI - Hollow polymer shells from biological templates: fabrication and potential applications. AB - Three-dimensional ultrathin polymer shells have been produced by a combination of step-by-step adsorption of polyelectrolytes on glutaraldehyde-treated human erythrocytes and subsequent solubilization of the cytoplasmatic constituents by means of a deproteinizing agent. The obtained hollow films preserve both the size and shape of the templating cells. This opens a pathway for the fabrication of polymeric capsules within a wide range of size and shape by using various biological templates. They may have exciting potential applications, such as templates for nanocomposites, as containers for a large class of materials, or as cages for chemical reactions. The thickness of the films can be adjusted over a large range: from a few nm up to several tens of nm. The polymer shells are permeable to small molecules and ions but not to macromolecules. An increase in the ionic strength of the solution up to 100 mmol make the capsules permeable for proteins. Permeability and conductivity studies have provided evidence that the adsorption of lipids on polyelectrolyte layers is a means of producing capsules with controlled permeability properties. 6-Carboxyfluorescein and Rhodamin 6G were precipitated within the capsules. PMID- 12561321 TI - Stereospecificity and enantioselectivity in the binding of the platinum(II) complex [PtCl2(tmdz)] (tmdz = 5,5,7-trimethyl-1,4-diazacycloheptane) to dinucleotides and oligonucleotides. AB - The two stereoisomers formed on reaction of each of the enantiomers of [PtCl2(tmdz)] with d(GpG) have been identified by using one- and two-dimensional 1H NMR spectroscopy. For both isomers formed with the R enantiomer the 3'-H8 shifts are downfield from those for the 5'-H8. For the S enantiomer the reverse is observed, showing that the bulky tmdz ligand determines the pattern of shifts. Models of these isomers generated by molecular mechanics show that the bulky tmdz ligand limits the rotation of the guanine bases and enforces right-handed (R2) canting for both isomers formed by the R enantiomer and left-handed (L1) canting for those formed by the S enantiomer. The pattern of H8 shifts is the opposite to that expected for these cantings; this suggests that other factors may play a role in determining these shifts. The interactions between the tmdz and d(GpG) ligands are also shown by molecular mechanics and the broadness of the H8 NMR signals to influence the tendency of the coordinated guanine bases to rotate about their Pt-N7 bonds. Reaction of each of the enantiomers with a 52 base-pair nucleotide, with a total of six GpG binding sites, resulted in the formation of only one of the stereoisomers in each case, the first reported case of complete stereoselectivity, or stereospecificity, in the reaction of Pt complexes with DNA. The observed stereoisomers were identified by comparison with the properties of the d(GpG) complexes. Molecular mechanics models of the adducts with duplex DNA show that the nonformation of one stereoisomer is consistent with the steric bulk of the tmdz ligand preventing closure from the monofunctional adduct to the bifunctional adduct. Enantioselectivity is also observed in that the R enantiomer forms more monofunctional adducts than bifunctional (59:41), whereas the S enantiomer forms more bifunctional adducts (27:73). The origins of this enantioselectivity must be at the level of monofunctional adduct formation and this has been investigated by molecular mechanics modelling. PMID- 12561322 TI - Morphological control of helical solid bilayers in high-axial-ratio nanostructures through binary self-assembly. AB - Mixed molecular species of cardanyl glucoside derived from renewable resources provide nanotubes upon self-assembly in water, while the saturated homologue generated a twisted fibrous morphology. The cardanyl glucoside mixture was fractionated into four individual components in order to study their contribution to the nanotube formation. The rational control of self-assembled helical morphologies was achieved by binary self-assembling of the saturated and monoene derivatives. This method can generate a diversity of self-assembled high-axial ratio nanostructures (HARNs), ranging from twisted ribbons and helical ribbons to nanotubes. PMID- 12561323 TI - The gas-phase route from Cp*2P6 to neutral hexaphosphorus. AB - Density functional theory has been applied to gain insight into the fragmentation and redox behavior of CpnP6+/0 and Cp*nP6+/0 cations and neutral species (n = 1, 2) in the gas phase. Particular attention is paid to the previously reported generation of neutral hexaphosphorus upon high-energy collisions of the Cp*P6+ cation. Theory provides an explanation for the experimentally observed effect that collisional electron transfer to the Cp*P6+ cation is negligible in that the associated Franck-Condon factors are predicted to be unfavorable. In contrast, dissociation of Cp*P6+ into Cp*(+) + P6 has a relatively low energy demand, thereby accounting for the efficient formation of neutral P6 in the gas phase. Theoretical exploration of the parent compound Cp2P6 reveals that the unsubstituted cyclopentadienyl ligand is much less suitable in this respect, thereby sustaining the previous suggestion that Cp* is a particularly good leaving group. PMID- 12561324 TI - [Cytotoxic activity of dammarane triterpenoids from birch leaves]. AB - Cytotoxic activity of dammarane triterpenoids isolated from beach leaves was studied. These substances differ from the native ginseng genin (20(S) protopanaxadiol) by the number, location, or configuration of OH-groups. Using fertilized egg cells of sea urchin Stronglyocentrotus intermedius we demonstrate that the orientation of C-3 OH-group has no effect on cytotoxic activity of triterpenoids as well as a higher activity of a triterpenoid with 3 alpha,12 beta OH as compared to a C-3 ketone but lower activity as compared to a triterpenoid with 3 alpha,17 alpha-OH. Depending on the number of OH-groups the cytotoxic activity of triterpenoids decreases in the row: tetraol > pentaol > triol. Dammar 24-ene-3 alpha 2 beta,17 alpha,20(S)-tetraol (compound IV) is cytotoxic for the Ehrlich ascite carcinoma cells and this effect is additive to cytotoxic activity of anthracycline antibiotic carminomycin in vitro. Compound IV changes the permeability and microviscosity of the tumor cell membranes. PMID- 12561325 TI - [Ecdysterone modulates antitumor activity of cytostatics and biosynthesis of macromolecules in tumor-bearing animals]. AB - The influence of therapeutic and half doses of cisplatin and adriamicin combination with the anabolic drug ecdisteron (20-hydroecdison) on development of subcutaneously and intraperitonially transplanted P388 and L1210 leukemia and metastasizing B16 melanoma was studied. Ecdisteron significantly stimulated the chemotherapeutic effect of low doses of the cytostatics: inhibition of tumor growth, mice survival rate, their lifespan, and the antimetastatic activity index were comparable or better than after therapy with high doses of the antitumor drugs. The influence of high and low doses of cisplatin and its low dose in combination with ecdisteron on the dynamics of protein and DNA biosynthesis in the liver, pancreas, thymus, spleen, and adrenals of tumor-bearing mice were also studied. Although the therapeutic effect of 4 mg/kg cisplatin by activated protein biosynthesis and DNA repair is comparable or better than that of its low dose (2 mg/kg) in combination with ecdisteron, in terms of chemotherapy the combination looks preferable since the therapeutic dose of cisplatin is toxic for the intact tissues. PMID- 12561326 TI - [Changes in sensitivity of lymphocytic leukemia P388 cells to oxidative stress and platidiam upon tumor growth]. AB - We studied the effect of oxidative stress induced by hyperoxia, hydrogen peroxide, or menadione on mouse leukemia P388 cells at early (4 days) and late (7 days) stages of tumor growth. Oxidative stress proved to inhibit cell division and to induce apoptosis. Seven-day leukemia cells feature lower proliferative potential and higher sensitivity to oxidative stress and platidiam. PMID- 12561328 TI - [Specific features of microsporogenesis in the hemp of the Shu valley]. AB - Specific features of microsporogenesis were studied in the narcotic drug containing hemp of the Shu valley. Nuclear divisions were investigated from the moment of appearance of the archesporial tissue until the formation of mature pollen grains. A special attention is paid to the detailed cytological analysis of meiosis and defects at all meiotic phases. The defects at anaphase 1 and anaphase 2 did not affect the karyotype constancy and their absence at the tetrad stage can be considered as an evidence of cytological stability of the studied material. PMID- 12561327 TI - [Expression of regulatory homeobox genes during retina regeneration in adult newts]. AB - We studied molecular-genetic mechanisms of retina regeneration in amphibians and, specifically, expression of the homeobox genes Pax6, Prox1, and Six3 in normal development and during retina regeneration in the newt. Based on the structural analysis of genes in closely related amphibian species, primers were constructed that flank certain regions of these genes. PCR fragments of calculated length were obtained. The affiliation of PCR products to the above genes was confirmed by sequencing. A comparative PCR analysis of expression of Pax6, Prox1, and Six3 was carried out in the native and regenerating newt retina, which allowed estimation of the level of expression. cDNA libraries obtained from the native and regenerating retina were used as templates. The libraries were preliminary standardized according to glyceraldehydes-3-phosphate dehydrogenase, an enzyme of general cell metabolism. The genes we studied were expressed in both native and regenerating retina. The level of Pax6 and Prox1 expression increased during regeneration, while that of Six3 decreased. The decrease in the level of Six3 expression could be due to antagonistic interrelations of Prox1 and Six3. The changed level of Prox1 and Six3 expression is a new fact and requires further studies. The interactions between these and other regulatory genes and localization of their expression in the cells of native and regenerating retina will be studied using in situ hybridization and immunohistochemistry. PMID- 12561329 TI - [Influence of exogenous salicylic acid on the level of phytohormones in tissues of Phlox paniculata and Phlox setacea leaves with special reference to resistance against the powdery mildew causative agent Erysiphe cichoracearum D.C. f. phlogis Jacz]. AB - We studied the effects of exogenous salicylic acid on the level of endogenous cytokinins and abscisic and salicylic acids in the tissues of leaves of phloxes contrasting in resistance against the powdery mildew causative agent: susceptible Phlox paniculata L. and resistant Ph. setacea L. Studies were carried out under the conditions of biotic stress. The initial level of salicylic and abscisic acids and cytokinins is the highest in the resistant phlox species. After treatment with salicylic species, the total level of cytokinins and endogenous salicylic acid increased in both species. When the treated phlox species were infected by the powdery mildew causative agent, the level of abscisic and salicylic acids increased in the susceptible Ph. paniculata, while that of cytokinins increased in the resistant Ph. setacea. The role of salicylic acid in the induction of plant defense reactions against phytopathogens is discussed. PMID- 12561330 TI - [Mitochondria respiration in developing sunflower seeds]. AB - We studied the oxidative and phosphorylative activity of mitochondria in the seeds of three sunflower cultivars (Polevik, Peredovik, and Yubileinyi) during development of the seed embryo within 1 to 54 days after flowering. The rates of succinate oxidation by the mitochondria were 1.5-2 times those of malate or alpha ketoglutarate oxidation. The ratio of substrate oxidation rates underwent changes during the seed growth. The differences were recorded between cultivars as concerns the times when the maximum oxidation rates were reached. Oxidation was coupled with phosphorylation during the entire period of seed development: the value of respiratory control after Chang changed from 1.4 to 7. By the time of transition to maturation, the rates of oxidation of both substrates and the values of respiratory control and ADP/O decreased. The results we obtained suggest that by days 13-15 of seed embryo growth, the rate of ATP production decreases upon oxidation of Krebs cycle products. PMID- 12561331 TI - [Detection of hepatoid glands and distinctive features of hepatoid acinus]. AB - In the 1920s-1930s, skin glands of a new type, hepatoid glands, were described in 13 mammal species (Rodentia, Canidae, and Bovidae). The hepatoid glands resemble sebaceous glands in their morphology, bur radically differ from them in specific structure of the acinus and another type of secretion. Later, these data either could not be confirmed or were considered insignificant and the hepatoid glands were described as modified sebaceous glands, glands with uncertain function, or modifications of epidermis. Based on the studies of various hepatoid glands in 22 species of Carniviora and Artiodactyla, the authors described in detail the characteristic features of the hepatoid acinus, which allow a precise discrimination of hepatoid and sebaceous glands. Extracellular secretory canaliculi have been described in the hepatoid glands, as well as the richness of hepatoid glands in protein, distribution of hydrophobic lipids in certain hepatoid glands, and formation of excretory ducts and cysts. The hepatoid glands are a source of great amounts of protein secreted in the merocrine way; the secretory substance of some of these glands has a strong odor. PMID- 12561332 TI - [The mechanisms underlying the formation of systemic hemodynamic reactions: role of initial vessel tone and arterial pressure]. AB - A review of the experimental materials describing the relationships between the initial (background) vascular tone and arterial pressure, on the one hand, and reactivity of the arterial system, on the other. Experiments were carried out on anesthetized rats under arterial hypotension caused by orthostasis or vasoactive agents, such as papaverin and sodium nitroprusside, and arterial hypertension resulting from volume expansion. It has been proposed that these relationships are based on changes in the amplitude of transmural pressure in arterial vessles. PMID- 12561333 TI - [A marker of early stages of diabetic retinopathy]. AB - A comparative estimate of the hormonal status and coagulatory activity was carried out in patients with diabetes mellitus at the functional and subclinical stages of diabetic retinopathy and in those without signs of diabetic retinopathy. In all patients, the blood level of hormones was elevated, while in patients with functional and subclinical stages of diabetic retinopathy, the changes in hormonal status were accompanied by increased local and systemic potentials, disturbed microcirculation, and decreased functional activity of the retina outer layers. The increased hormone level affected the hemostatic potentials and induced their elevation at a certain stage. The reliably increased local hemostatic potential was one of the first signs of pathological action of the increased hormone level, while the increase of systemic hemostatic potential was unreliable. The combination of elevated blood level of hormones and coagulatory activity of the tear fluid is a marker fore revealing the group of risk for development of diabetic retinopathy in patients with diabetes mellitus. PMID- 12561335 TI - [Energy bases and scale of the production-destruction balance in the Black sea: photosynthesis and flows of C(org), O2 and CO2]. AB - The scale of production-destruction biotic balance in the Black Sea ecosystem, flows of organic carbon, carbon dioxide, and oxygen, and rate and volume of organic matter input in the Black Sea sediment as a source of formation and accumulation of fossilized hydrocarbon fuel minerals are estimated on the basis of the current concept of efficiency of solar radiation utilization by photosynthesis. The volume of "excessive" oxygen formed in the ecosystem and its contribution to global oxygenation of the atmosphere are shown. PMID- 12561334 TI - [Anticoagulant, fibrinolytic and antithrombotic effects of the heparin-insulin complex]. AB - Formation of heparin-insulin complex at the 1:10 molar ratio of the components has been demonstrated by spectral methods. The derived complex had anticoagulant, antithrombotic, and fibrinolytic properties of non-enzymatic nature in vitro. Intravenous injection of the complex in the animals (rats) increased anticoagulant and fibrinolytic background and at the same time decreased the plasma coagulation factors fibrinogen and factor XIIIa in the bloodstream. We propose the heparin-insulin complex as a promising antithrombotic drug. PMID- 12561336 TI - [Morphofunctional features of nutrition of certain predatory gastropods]. AB - Spectra and modes of nutrition as well as morphobiochemical adaptations to nutrition in certain predatory gastropods have been considered. Hemoglobin content in the radular tissues as a function of nutrition mode was assessed. PMID- 12561337 TI - [Using biological, hydrochemical and hydrological data on freshwater in Russia and neighboring countries for constructing a computerized information system]. AB - Raw hydrobiological (population, biomass, and numbers of phytoplankton, zooplankton, zoobenthos, periphyton, and bacterioplankton species) and physicochemical indices of ecosystems are integrated into information system "Ecology of Freshwater in Russia and Neighboring Countries." This system is dedicated to environmental control by biotic indices in former USSR countries. The information system is realized as an Ecograde application relying on Microsoft Access software as well as a web site. PMID- 12561338 TI - [Ecology of testate amoebae from oligotrophic peatlands: specific features of polytypic and polymorphic species]. AB - Relationships between species abundance and water table depth and soil moisture have been modeled using weighted averaging and species niches have been calculated as optima and tolerance for these parameters. Niche separation of closely related taxa has been examined in detail and it has been shown that there is often a gradient of hydrological preference within each taxon. Wet to dry gradients include those found in the Trigionopyxis arcula group (T. arcula var. major > T. arcula > T. minuta), Assulina-Valkanovia group (A. seminulum > A. muscorum > V. elegans), and Trinema lineare group (T. lineare var. truncatum/T. lioneare > T. lineare var. terricola), all of which are associated with a large to small size gradient. In addition, spined forms within the Euglypha and Placocista genera have been shown to consistently occur in wetter habitats than glabrous forms of those with shorter spines. A conclusion has been drawn that palaeoecological studies should cover the lowest taxa possible within these groups to maximize the ecological indicator value of the assemblages recorded. PMID- 12561339 TI - [Testaceans (Testacea, Protozoa) of taiga soils in Western Siberia (Surgut woodland)]. AB - We studied population structure of testaceans of automorphic and hydromorphic soils in the middle taiga of Western Siberia. Rich testacean fauna has been revealed. It was represented to a similar extent as compared to the soils of Ciscaucasian Russia. The bryophilic group was the most typical for the region, since its representatives inhabited the predominating hydromorphic soils of bogs. The typical species included Amphitrema flavum, Heleopera petricola, Placocista spinosa, Trigonopyxis arcula, Centropyxis laevigata, Nebela tincta, Arcella catinus, as well as other moss forms. Flood plain proved to be the richest by testacean species variability among the studied habitats. The set of species here included both representatives of watershed and typical aquatic forms present due to the flood conditions of a given area. The highest quantitative indices of the testacean complexes were revealed in the forest area soils. Quadrulella quadrigera Deflandre, 1936 has been found in a flood plain area for the first time in Russia. PMID- 12561340 TI - [Influence of different ecological groups of earthworms on the intensity of nitrogen fixation]. AB - The results of studying nitrogen fixation in organic substrates processed by different ecological groups of earthworms suggest that the earthworms Aporrectoidea caliginosa actively stimulate nonsymbiotic nitrogen fixation. In this respect, they exceed the manure worms Eizenia foetida tens of times due to the formation in the organic substrate of conditions favorable for the nitrogen fixing bacteria, and namely: low content of nitrogen easily accessible for microorganisms, changes in the structure of the microbial community of the substrate in favor of non-spore forms of bacteria, and suppression of the growth of saprophyte bacilli, the main competitors of nitrogen fixing bacteria for carbon nourishment sources. PMID- 12561341 TI - [Respondence to electricity in the muscles of rat's jaw]. AB - 15 male Wistar rats of 8 weeks old were used in this experment. After deeply anesthesia, the masseter muscles, digastric muscles, lateral pterygoid muscles were stimulated in the apartment that was made by us. The curves that express the relations of force-electronic stimulates were recorded and the constitutive equations of these muscles were given. When a single electronic signal stimulates the muscle, the respondence of the muscles can be expressed as F = A(e-alpha t - e-beta t) and the constant A, alpha, beta were determined. When the frequency of the electronic stimulation was higher than 3 Hz, the respondence was expressed as F = Ce-gamma/t + Dsin omega t and the constants C, D and gamma were determined. When the frequency of the electronic stimulation was thirty or higher, the tetanic convulsion occurred. PMID- 12561342 TI - [Simulation of the deformation of the endothelial cell under a shear flow]. AB - The coupling between the endothelium and blood flow is an important biomedical problem and has drawn extensive research. Endothelial cells are known to adapt their shapes and functions in response to applied shear flow. Shear Stress being regarded as a primary triggering signal for cellular remodeling, it is important to understand the interaction mechanism between applied shear flow and endothelial cells. In present study we have established a theoretical model to simulate the coupling between the deformation of an endothelial cell and applied shear flow. A two dimensional computational fluid dynamic (CFD) is conducted to determine the local distributions of mechanical stress and pressure on cell surface. Our results show that: (1) the deformation of endothelial cell changes with alpha (corresponding to the shear stress imposed on cell surface by flow fluid). When alpha is greater than 0.021, the cell deformability increases greatly; (2) the distributions of stress and pressure on cell surface are not uniform, but the maximal shear stress and displacement are always at the top point of the cell. Meanwhile, we have measured the deformation of cultured human aortic endothelial cells (HAECs) exposed to shear flow by using a flow chamber. We found that the numerical results are well consistent with those of experiment. These results suggest that the non-uniformity distributions of mechanical stress and pressure on cell surface may play a particular role in the mechanism of cell activation and in the regulation of endothelial cells functions (modification of cytoskeleton, distributions of adhesion molecules, etc.). The present study offers a framework to facilitate the development of a comprehensive dynamic model for endothelial cells. PMID- 12561343 TI - [Study on relationship between hemocompatibility and protein adsorption for DLC]. AB - In this paper, the adsorption of human serum albumin(HSA), human serum fibrinogen (HFG) and human serum immunoglobin(IgG) on diamond like carbon film(DLC) has been studied in comparison with diamond film (DF) and graphite. The isothermal adsorption of the protein solution with single component and the competitive adsorption of binary protein system have been investigated by radio isotope 125I labelling method. Results showed that (1) the adsorptive amounts of three proteins on three material surfaces are all increased with the increasing concentration of protein solution, then the adsorption tends to reach an equilibrium; (2) the adsorptive amounts of three protein on graphite far exceed that on DLC and DF; (3) the adsorptive amounts of HSA on DLC are more than that on DF, while the adsorptive amounts of HFG and IgG on DF and graphite are apparently more than that on DLC; (4) the differences among the adsorptive amounts of three proteins on DLC are small, but adsorptive amounts of HFG and IgG on DF and graphite are much more than that of HSA; (5) the relative competitive adsorption ability of three proteins on DF and graphite is HFG > IgG > HSA, but on DLC, the sequence is HFG approximately HSA > IgG. Comparing with HSA, HFG has no apparent competitive adsorption superiority to DLC. These results indicate that there is no apparent difference for the adsorption of three human serum proteins on DLC, but the adsorption of HFG and IgG on DF and graphite takes precedence of various degrees. It probably makes a rational explanation for the result of hemocompatibility tests in vitro that DLC is superior to, DF and graphite. PMID- 12561344 TI - [Study on the sealing capability of a calcium phosphate root canal filling material]. AB - To evaluate the sealing potential of self-designed root canal filling material made of calcium phosphate cement (alpha-TCP/TTCP, CPC), the apices of root canals of six adult dogs were purposely perforated and enlarged up to the No 40 instrument. Then CPC was used to fill the root canal. Mean while either calcium hydroxide (Ca(OH)2) paste or hydroxyapatite (HA) paste was used as control. The animals were killed at 4, 12, 20 weeks postoperatively. The different materials about ways of apical closure, restoration periapocal tissues and adaptability to the dentinal surface were observed by histomorphology and scanning electron microscopic. This study revealed that CPC had excellent biocompatibility and adaptability to the dentinal wall. Its osteoconduction can promote the formation of calcific barriers and healing of periapical tissue. The apex can be closed completely. Compared with the control pastes it has advantages of ease of manipulation and better sealing capability. The results showed that CPC could be used as a root canal filling material for pupless teeth with open apex and destructive periapical tissue. PMID- 12561345 TI - [The impedance frequency response of the human skull]. AB - The human skull impedance was quantitatively measured by means of Frequency Response Analyzer and its software. The impedance-frequency response curves of human skull were obtained. By analyzing the curves, we found that the characteristic frequency of human skull should be around 10 KHz. PMID- 12561346 TI - [Preparation of interferon-alpha-loaded poly-L-lactic acid lamellar particles]. AB - Lamellar particles(lamellae) were prepared by non-solvent precipitation from crystalic poly-L-lactic acid (PLLA). The PLLA lamellae exhibite a diamond or stepped irregular shape with a size range between 3-5 microns. Prepared without any surfactants and dispersing agents, the lamellar particles have clean surface, which is advantageous for the adsorption of proteins. The PLLA lamellar particles adsorbed protein cytokine interferon-alpha (IFN-alpha) with an adsorption efficiency of > or = 95%. The release of loaded IFN could continue for more than 10 days. The cell incubation experiments showed that the PLLA lamellar particles were easy to be phagocytosed by macrophages. The immunological experiments showed that the biological activity of IFN-alpha loaded on the PLLA lamellar particle was effectively retained. PMID- 12561347 TI - [An experiment study on repairing residual dental root with a horn-shaped orifice by different materials]. AB - When a traditional method is used to repair residual dental root with horn-shaped orifice, this kind of dental root often has to be extracted since the success rate in clinic is very low. The main reason is that the stress often concentrates on the neck and root apex, therefore the wall at the orifice is too thin to bear this kind of pressure. A new repairing method is proposed trying to preserve the residual dental root with horn-shaped orifice. Then compare the anti-fracture ability of this kind of tooth with that of no repairing. The results show that it is very effective to prevent the fracture of residual dental root repaired by chemical hard synthetic resin followed by making the post and core. However, there is no obvious difference between non-repaired and residual dental root groups repaired by traditional glass ionic adhesives. The results also provide the great possibility to preserve the residual dental root with horn-shaped orifice in clinic. PMID- 12561348 TI - [The influence of vagus nerve stimulation on NMDAR1 mRNA and GABAAR alpha 1 mRNA in thalamic reticular neucus of pentylenetetrazole-induced epileptic rats]. AB - To study the antiepileptic mechanism of vagus nerve stimulation (VNS), we used the methods of in situ hybridyzation and image analysis to detect the expression of NMDAR1 mRNA and GABAA receptor alpha 1 subunit mRNA (GABAAR alpha 1 mRNA) in the thalamic reticular nuclus. The results show that the NMDAR1 mRNA expression of rats administered pentylenetetrazole(PTZ) is higher than that of control group. By treating with VNS, it decreased. On the contrary, the expression of GABAAR alpha 1 mRNA in the thalamic reticular nuclus of PTZ group rats is lower than that of control group. For rats treated with VNS, it increased. Therefore, it is concluded that VNS may reduce the excitability of cerebral cortices by depressing the activities of glutamic acid receptors (GluR) and by promoting the activities of gamma-aminobutyric acid receptors(GABAR) in thalamic reticular nuclus. So the formation and development of seizures are inhibited. PMID- 12561349 TI - [Design of MC-III low frequency pulsed strong magnetic fields generator]. AB - In this paper, We designed and accomplished a low frequency pulsed strong magnetic fields generator, which provides a pulsed magnetic field with the intensity range from 0.1-2.5 T and the adjusted time interval of pulse. This device is easy to operate and performs reliably. It can work steady for a long time and has been successful used in the experiments of biological effects of electromagnetics. PMID- 12561350 TI - [Study on adhesion of platelets to surface of copoly(benzyl glutamate-hydroxy ethyl glutamine)]. AB - Copoly (benzyl glutamate-hydroxy ethyl glutamine) were prepared by partially aminolysis of poly benzyl glutamate with hydroxyl ethylamine at 60 degrees C for predeternined period. As aminolysis was proceeding, the content of hydroxy ethyl glutamine in copolymer increased and the yield of aminolysis decreased. When aminolysis time reached 16.5 hours, a copolymer with 0.357 mole fraction of hydroxy ethyl glutamine was obtained. After 16.5 hours a soluble yellow viscous product of reaction was obtained. The aminolysized specimens displayed more swelling degree in water, that is, more hydrophilits. The meassurement of solid liquid contact angles showed that an increase in critical surface tention with content of hydroxy ethyl glutamine in copolymer was observed. Likewise a obvious increase in polar component and a slight decrease in dispersive of surface free energy, thus an increase in total surface free energy with content of hydroxy ethyl glutamine was found. The platelet adhesion and deformation test indicated that less platelets were adhered to surfaces of all aminolysized specimens than that of both poly benzyl glutamate and polydimethylsilicone. The least adhered platelets on surface of copolymer with 0.133 mole fraction of content of hydroxy ethyl glutamine were observed. The partiall aminolysis of poly benzyl glutamate films is an effective method to improve its hydrophility and antithrombogenicity. PMID- 12561351 TI - [HAP-glutin-acetic acid coating on degradable polyamides implant]. AB - Hydroxyapatite (HAP) formation on the surface of the degradable polyamides implant by a coating technology was studied. A suspension of HAP-glutin-acetic acid was prepared and brushed on the surface of the implant at room temperature. Scanning electron microscopy (SEM), X-ray diffractometry (XRD) and Fourier transform infrared spectroscopy (FT-IR) were employed to study the composition and the microstructure of the coating. The results obtained indicate that the main composition of coating is crystal HAP, its structural formula is Ca10(PO4)6(OH)2. So the coating has bioactivity. The coating has also small pores, which can stimulate bone regrowth and adsorption of other bioactive materials or mediine. This technology is an effective method to improve medical performance of the polyamides implant. PMID- 12561352 TI - [The research on computer modeling for artificial bio-valve]. AB - In this paper, the structure parameters and the equations to calculate these parameters are presented for the model design of the artificial bio-valve on the basis of heart anatomy, shell theory and the optimal flow of heart valve. No-root bio-valve is designed and manufactured according to the above theoies and technology. Its perfect property has been confirmed by measurement in vitro and clinical application in vivo. PMID- 12561353 TI - [The theoretical calculation for optimal doping content of blood compatible titanium oxide films]. AB - The quantitative theory of optimal doping content of transparent conductive films is introduced in this paper. In this theory, the relationship among the physical properties and crystal structure, preparation method and doping content is set up. The parabola equation that can be fixed to test curve and that has the reliable physical meaning is given. The extreme value of this equation confirms the quantitative relationship between the optimal doping content, the crystal structure and the preparation method. This an expression of the optimal doping content is obtained. The experimental results of doping modified for blood compatibility of titanium oxide films are analyzed. Using this expression to calculate the Ta5+ optimal doping content in blood compatibility of titanium oxide films. PMID- 12561354 TI - [Comparative study on using TTCP and CTCP ceramic artificial bone for repairing segment defect of long bone]. AB - To develop ideal ceramic artificial bone for accelerating the process of repairing segment defect of long bone, tubing tricalcium phosphate (TTCP) and column tricalcium phosphate (CTCP) ceramic artificial bones were implanted separately into radial defects of 1 cm in adult rabbits. Specimens were taken in 4 and 12 weeks. The samples were examined by gross histomorphological observation, quantitative measurement of new bone formation and bending strength. The amount of new bone formation in TTCP was more than that in CTCP at 4 and 12 weeks postoperatively. At 12 weeks after operation, the grafted materials were bonded closely with host bone. The degradation rate of TTCP was faster than that of CTCP, and the bending strength of TTCP was stronger than that of CTCP(P < 0.01). The results showed that the design of TTCP is relatively rational and TTCP could accelerate the repair of segment defect of long bone more rapidly than CTCP. TTCP is an ideal artificial bone material. PMID- 12561355 TI - [Corrosion rate measurements of biomedical TiNi shape memory alloy and cobalt alloys]. AB - The corrosion rates of TiNi, CoCrNiW and CoCrNiMo were measured in Tyrode's solution with potentiodynamic linear polarization, fore-point weak polarization, Cao Chunan weak polarization, transient linear polarization and atomic absorption spectroscopy. Results indicated that corrosion rates of these three alloys were very low due to their excellent corrosion resistance and the corrosion resistance of CoCrNiMo was the best. Corrosion rates of TiNi, CoCrNiW and CoCrNiMo were 0.691, 0.0595, 0.0490 micron/a and 0.0528, 0.0383, 0.0387 micron/a, respectively. The results measured by the first three methods were about ten times of those by the latter two methods, this was related to the applicability of each method and the alloy surface state. Transient linear polarization technique can determine low corrosion rate conveniently and quickly. Atomic absorption spectroscopy method, determining directly the concentration of ion in solution, and thus provide reference for material biocompatibility. And these two methods are properly used in measuring corrosion rates for biomedical materials. PMID- 12561356 TI - [Improved design of permanent maglev impeller assist heart]. AB - Magnetic bearing has no mechanical contact between the rotor and stator. And a rotary pump with magnetic bearing has therefore no mechanical wear and thrombosis due to bearing. The available magnetic bearings, however, are devised with electric magnets, need complicated control and remarkable energy consumption. Resultantly, it is difficult to apply an electric magnetic bearing to rotary pump without disturbing its simplicity, implantability and reliability. The authors have developed a levitated impeller pump merely with permanent magnets. The rotor is supported by permanent magnetic forces radially. On one side of the rotor, the impeller is fixed; and on the other side of the rotor, the driven magnets are mounted. Opposite to this driven magnets, a driving motor coil with iron corn magnets is fastened to the motor axis. Thereafter, the motor drives the rotor via a rotating magnetic field. By laboratory tests with saline, if the rotor stands still or rotates under 4,000 rpm, the rotor has one-point contact axially with the driving motor coil. The contacting point is located in the center of the rotor. As the rotating speed increases gradually to more than 4,000 rpm, the rotor will detache from the stator axially. Then the rotor will be fully levitated. Since the axial levitation is produced by hydraulic force and the driven magnets have a gyro-effect, the rotor rotates very steadly during levitation. As a left ventricular assist device, the pump works in a rotating speed range of 5,000-8,000 rpm, the levitation of the impeller hence is ensured by practical use of the pump. PMID- 12561357 TI - [Observation of the antithrombotic property of a new prosthetic heart valve in sheep]. AB - To observe the antithrambotic property of a new prosthetic made double bileaflet valve of all-pyrolytic carbon. METHOD: Fifteen sheeps underwent mitral valve replacement (MVR) with this new mechanical valve. Each animal was placed on cardiopulmonary bypass. All of the sheep were given in an anticoagulation protocol and followed for three months. Final studies were performed on all animals surviving for more than 12 weeks. RESULT: Ten sheeps died within 48 h after operation, five sheeps survived. The long-term survivors were killed forautopsies, two sheeps were killed in the 1st week after operation, one in 12th weeks and two in 2.5 years. Pathological examination of 10 sheeps which died early after operation revealed that there was no either evidence of thromboembotic phenomenon, or any failures of the mechanical valve. Autopsy studies in the five long-term survivors revealed that the sewing ring was covered by a thin layer of microthrombi consisted most of platelets and fibrin in 1th week after operation. The sewing rings was covered by a tissue layer, with partial organized micro thrombi and endothelialization in 12th weeks after the operation. Two and half years after operation, the complete endothelialization presented around the sewing ring without restriction of bileaflet motion. The tissue ingrowth tends to stop at the pyrolytic carbon-fabric interface on both surfaces of left atrium and ventricle. This study demonstrated that the new prosthetic heart valve had not promoted thrombosis and thrombembolism. PMID- 12561358 TI - [Method of multi-resolution 3D image registration by mutual information]. AB - Maximization of mutual information is a powerful criterion for 3D medical image registration, allowing robust and fully accurate automated rigid registration of multi-modal images in a various applications. In this paper, a method based on normalized mutual information for 3D image registration was presented on the images of CT, MR and PET. Powell's direction set method and Brent's one dimensional optimization algorithm were used as optimization strategy. A multi resolution approach is applied to speedup the matching process. For PET images, pre-procession of segmentation was performed to reduce the background artefacts. According to the evaluation by the Vanderbilt University, Sub-voxel accuracy in multi-modality registration had been achieved with this algorithm. PMID- 12561359 TI - [Antigen expression and the biomechanical characteristics of the biologic blood vessel matrix]. AB - To explore the changes of the antigen expression and the biomechanical characteristics of blood vessel in Banna little ear pig before and after trypsin treatment, and provide data for xenotransplantation and pig vessel using for tissue engineering. Geometric morphology and microstructure of pig cartoid artery were stuided quantitatively by histologic method and computer image analysis. The relationship between pressure and diameter was observed at different period of time before and after trypsin treatment. Affinity-immunohistochemistry assay was conducted to detect the expression of xenoantigens (alpha-Gal). The results showed that alpha-Gal antigen is only expressed in vascular endothelial cellsouly. There is no significant difference in blood vessel compliance. These demonstrate that the antigenicity of pig carotid artery is significantly reduced, however, the mechanical characteristics did not change significantly. We suppose that pig vessels treated by trypsin can be used as the substrate material for vascular tissue engineering. PMID- 12561360 TI - [Correlation of length of vascular segment in vitro with endothelin-1 metabolism of endothelial cells--study on accumulative effect of tensile stress in the upper endothelial cell membrane. III]. AB - A few of flow experiments were utilized to verify a theoretical hypothesis proposed by Fung and coworker which showed that the tensile stress in the upper cell membrane of the vascular endothelium could accumulate upstream against the direction of blood flow. Endothelial cells from replicate human umbilical vein segment (HUVSEC) in vitro with length of 11 cm and 21 cm were exposed to the same pulsatile laminar shear stress averaged of 0.12 N/m2 for 42 hours. The average production rate of endothelin-1(ET-1), at 11 cm segment is 50% lower than that at 21 cm segment(16.93 +/- 0.89) vs. (26.13 +/- 1.79) pg/cm2.h respectively. The average production rate of ET-1 under pulsatile laminar flow was significantly higher than that under steady laminar flow. It showed that, high correlation of the length of HUVSEC with their ET-1 metabolism exists, suggesting that the tensile stress in the upper endothelial cell membrane could accumulate. PMID- 12561361 TI - [The computer simulation of magnetic resonance imaging]. AB - Based on the Bloch equation, we can describe the working principle of the magnetic resonance imaging system with mathematic model, the inputs are the three templates of appropriate rho, T1 and T2. In this paper, the simulation process is illustrated by spin echo examples of the signal exciting, phase coding, reading out and image reconstruction. The satisfactory results show that this method can be widely used in simulation researches. PMID- 12561362 TI - [Effects of sampling parameter variation on the complexity analysis of EEG]. AB - The algorithmic complexity and the approximate entropy of EEG were calculated and analyzed with different data points, different sample frequencies and different sample time duration. The results showed that under fixed sample frequency, the longer the data was, the more stable the complexity values were. With fixed sample time duration or fixed data point, lower sample frequency would be better both for EEG distinguishing and for computing time saving. PMID- 12561363 TI - [Study on micro-hardness and wear resistance of human tooth at different depth]. AB - Micro-hardness of human teeth was measured by micro hardnessmentry at different depths i.e. enamal, enamelo-dentinal junction and dentin. A simulated dental friction test was conducted of respectively in these three areas, opposing pure titanium ball under a modified fretting machine. The results showed that micro hardness is 320.40 +/- 22.77 HV of enamal, 232.26 +/- 32.31 HV of enamelo dentinal junction, and 61.17 +/- 9.42 HV of dentin (P < 0.05). Wear depth is 17.33 +/- 0.58 microns of enamal, 49.44 +/- 16.47 microns of enamelo-dentinal junction and 95.20 +/- 15.07 microns of dentin(P < 0.05). Wear resistance of enamal is much better than that of dentin's. This preliminary investigation should be very useful in clinical applications. PMID- 12561364 TI - [Effect of moulding and extruding conditions on mechanical properties of poly(D,L lactide) and MDI chain-extending poly(D,L-lactide)/hydroxyapatite composite]. AB - In this paper, poly(D,L-lactide) (PDLLA), MDI chain-extending poly(D,L-lactide) (PDLLA/MDI) and MDI chain-extending poly(D,L-lactide)/hydroxyapatite composite (PDLLA/HA/MDI) were prepared respectively and the effects of moulding and extruding conditions on their mechanical properties were also investigated. At the optimal conditions, bending strength of PDLLA and PDLLA/MDI is 35.1 MPa and 51.3 MPa, respectively, and their bending modulus is 2413.6 MPa and 1830.9 MPa, respectively. Bending strength of PDLLA/HA and PDLLA/HA/MDI is 31.2 MPa and 55.4 MPa, respectively, and their bending modulus is 1735.0 MPa and 2068.5 MPa, respectively. These results have shown that the mechanical properties of PDLLA/MDI and PDLLA/HA/MDI have enhanced significantly by MDI chain-extending. PMID- 12561365 TI - [Registration and 3D rendering of serial tissue section images]. AB - It is an important morphological research method to reconstruct the 3D imaging from serial section tissue images. Registration of serial images is a key step to 3D reconstruction. Firstly, an introduction to the segmentation-counting registration algorithm is presented, which is based on the joint histogram. After thresholding of the two images to be registered, the criterion function is defined as counting in a specific region of the joint histogram, which greatly speeds up the alignment process. Then, the method is used to conduct the serial tissue image matching task, and lies a solid foundation for 3D rendering. Finally, preliminary surface rendering results are presented. PMID- 12561366 TI - [Characteristics of tenocyte adhesion to biologically-modified surface of polymer]. AB - In this study we examined the in vitro characteristics of tenocyte adhesion to biologically-modified surface of polymer. Polylactic-co-glycolic acid (PLGA) 85/15 films were prepared by a solvent-casting technique. Each film was adhered onto the bottom of a chamber. The film was precoated with poly-D-lysine (PDL), and then coated with serum-free F12 medium containing various concentrations of fibronectin (FN), type I collagen (CN I), and insulin-like growth factor1 (IGF 1). The monoclonal antibodies (to FN and to CN I) with various dilutions were used to inhibit attachment of tenocytes to surface precoated with FN or CN I. Human embryonic tendon cells (HETCs) and transformed human embryonic tendon cells (THETCs) were used as the seeding cells. The system used for the measurement of adhesion force was the micropipette aspiration experiment system. The micropipette was manipulated to aspirate a small portion of the tenocyte body by using a small aspiration pressure. Then the pipette was pulled away from the adhesion area by micromanipulation. The minimum force required to detach the tenocyte from the substrate was defined as the adhesion force. The results showed that modification of FN or CN I by precoating significantly enhanced attachment of tenocytes to surface of polymer (P < 0.05). As antibodies to FN or CN I were added to a polymer film precoated with FN or CN I, the adhesion force decreased significantly (P < 0.05). We concluded that the specific adhesion forces of tenocytes to extracellular matrix adhesion proteins (FN and CN I) had coordinated action and showed good dependence on their precoating concentrations, and were inhibited by the antibodies to these adhesion proteins. Films precoated with IGF 1 strongly accelerated the adhesion of tenocytes to polymer. These results indicate that the specific adhesion of tenocytes to polymer can be promoted by coating extracellular matrix adhesive proteins and insulin-like growth factor1. It is of great importance to construct tissue-engineered tendon. PMID- 12561367 TI - [Microencapsulation of immortalized mandibular condylar chondrocytes]. AB - To explore the possibility of microencapsulation of chondrocytes in cartilage tissue engineering, immortalized manibular condylar chondrocytes (IMCCs) were microencapsuled by Alginate-polylysine-alginate (APA) method, according to air pressure shearing model. Phase contrast microscopy, trypan blue staining exclusion, cell number counting, HE staining and immunohistochemistry method were used to observe the morphology of the microencapsules, the growth character of cells, cartilage characteristics, and so on. The results showed that IMCC could survive and grow in microencapsule, and the viability rate of cells is more than 80 per cent. The diameter of microcapsule is 779 microns in average. The number of cell increased with time, and cells went into platform in about 20 days. Cells grew in clusters and cartilage specific proteoglycans and type II collagen were highly expressed. It was concluded that IMCC could form cartilage-like tissue within microencapsulation, implying that microencapsule technique might be applicable to cartilage tissue engineering. PMID- 12561368 TI - [Research on stability of hemocompatibility of DLC film/Ti6Al4V gradient material using image analysis method]. AB - The stability of hemocompatibility of Ti6Al4V alloy and DLC film/Ti6Al4V gradient material has been studied. The platelet consumption ratio of Ti6Al4V alloy to DLC film/Ti6Al4V gradient material has been measured respectively by digital image analysis method. It has been found that the platelet consumption ratio of Ti6Al4V alloy increased remarkedly about 50% after 2,000 m sliding tribological test under the lubrication of Hank's solution, while that of DLC film/Ti6Al4V gradient material almost didn't increase under the same condition, which means that the DLC film/Ti6Al4V gradient material has better stability of hemocompatibility. PMID- 12561369 TI - [Theoretical calculation and experimental study of membrane thickness of alginate (poly-L-lysine)-alginate microcapsules]. AB - Alginate-(Poly-L-Lysine)-Alginate(APA) microcapsules were prepared by Electrostatic Droplet Generator(EDG) technique and the thickness of microcapsule membrane, which was composed by polyelectrolyte complex, were studied in this paper. The theoretical formula was given for the measurement of membrane thickness of APA microcapsules by element analysis of membrane and calculation. The membrane thickness was 7-10 microns by theoretical calculation. On the other hand, the thickness of membrane was measured by SEM and optical microscopy and the results were 7 microns and 12 microns, respectively. The results showed that theoretical calculation is in good accordance with experimental determoination of mermbrane thickness and the membrane thickness of APA microcapsule is about 7-10 microns. The optical microscopy is an easy way to measure membrane thickness. PMID- 12561370 TI - [Effects of shear stress and lysophosphatidylcholine on adhesion molecules expression of endothelial cells]. AB - Extensive monocyte recruitment is an early phenomenon associated with the development of atherosclerotic lesion. Although the molecular mechanisms are not completely understood, monocyte recruitment into these early lesions may involve changes in endothelial adhesion for monocyte, in which adhesion molecules expressed by endothelial cell play an active role. In vivo, the function of endothelial cells is not only affected by the chemical factors, but also by the mechanical factors. The purpose of this article was to investigate the induction of adhesion molecules expression by synergistic effects of Lysophosphatidylcholine (Lyso-PC) and shear stress in cultured human umbilical vein endothelial cells (HUVEC). The surface expression of ICAM-1, VCAM-1 and E selectin on HUVEC induced by Lyso-PC(30 micrograms/ml) and shear stress(2.23, 4.20 dyne/cm2) were analyzed using flow cytometry. The results showed that: Compared with what were simultaneously exposed to shear stress and Lyso-PC, activating the cells with Lyso-PC prior to shear stress, or pre-conditioning the cells exposed shear stress prior to Lyso-PC incubation, a significantly higher expression of ICAM-1 and VCAM-1(P < 0.05) was resulted. HUVEC were exposed to shear stress and Lyso-PC at the same time or treated with each agonist alone, E selectin expression was not significantly different from the control group. However, a sequential action of the two stimuli significantly increased E selectin expression(P < 0.05). We concluded that: a sequential action of the shear stress and Lyso-PC induced an even greater expression of ICAM-1 and VCAM-1, thus it could be understood that the flows-hear stress in combination with endothelial activated by chemical factors may increase the ability of endothelial cells to recruit leukocytes even under the mechanical environment unfavorable for cell adhesion. PMID- 12561371 TI - [The effects of tensile strain and loading time on the secretion of IL-1 beta of human PDLF]. AB - This study aims at exploring the effects of tensile strain and loading time on the secretion of IL-1 beta of human periodontal ligament fibroblast. Five tensile strain values including 0%, 8%, 12%, 16%, 20% and three loading time including 24 h, 48 h, 72 h are set in this study. The prepared cell samples are mounted on the self-devised loading apparatus in vitro. The content of IL-1 beta in each sample was determined using double-antibody ELISA. The secretion amount of IL-1 beta per day is directly proportional to the loading time and tensile strain value in tensile strain group of 8%, 12%, 16%. The secretion amount of IL-1 beta reaches its maximum at tensile strain value of 16% in loading time groups. Loaded strain for 24 h and 48 h, the secretion amount of IL-1 beta at tensile strain value of 20% is obviously more than that at value of 0%, but the amount already begin to decrease apparently. Loaded strain for 72 h, the secretion amount of IL-1 beta decreases to a great extent that it is less than the amount at strain value of 0%. The tensile strain stimulates the secretion of IL-1 beta by human periodontal ligament fibroblast when the strain is under normal physiological extent, but the stimulation effect fades out as time goes on. PMID- 12561372 TI - [An algorithm of a wavelet-based medical image quantization]. AB - The compression of medical image is the key to study tele-medicine & PACS. We have studied the statistical distribution of wavelet subimage coefficients and concluded that the distribution of wavelet subimage coefficients is very much similar to that of Laplacian distribution. Based on the statistical properties of image wavelet decomposition, an image quantization algorithm is proposed. In this algorithm, we selected the sample-standard-deviation as the key quantization threshold in every wavelet subimage. The test has proved that, the main advantages of this algorithm are simple computing and the predictability of coefficients in different quantization threshold range. Also, high compression efficiency can be obtained. Therefore, this algorithm can be potentially used in tele-medicine and PACS. PMID- 12561373 TI - [Long-term culture of dermis-derived multipotent stem cells and the effects of collagen sponge on their growth in vitro]. AB - Autologous multipotent stem cells are most relevant cells for regenerative medicine and show prosperous future in the treatment of human diseases. Previous reports have indicated that multipotent stem cells (MSCs) can be obtained from bone marrow and adipose tissues. In this study, we proved that dermis may be another source of these cells. MSCs were isolated from the dermis of newborn rats one day old by adhesion competition and successive culture. These cells conserved the ability to differentiate to osteoblasts, chondrocytes and adipocytes by induction media containing dexamethasone. After long term of more than 6 months, till 25th generation, the cells still maintained the characteristics of stem cells: high activity of self-renewal and multipotency. Mixed collagen matrix from dermis could promote the growth of dermis-derived multipotent stem cells and collagen sponge stent could promote their three-dimensional growth in vitro. PMID- 12561374 TI - [PIV: a computer-aided portal image verification system]. AB - Portal image verification (PIV) is one of the key actions in QA procedure for sophisticated accurate radiotherapy. The purpose of this study was to develop a PIV software as a tool for improving the accuracy and visualization of portal field verification and computing field placement errors. PIV was developed in the visual C++ integrated environment under Windows 95 operating system. It can improve visualization by providing tools for image processing and multimode images display. Semi-automatic register methods make verification more accurate than view-box method. It can provide useful quantitative errors for regular fields. PIV is flexible and accurate. It is an effective tool for portal field verification. PMID- 12561376 TI - [Evaluation of the performance of the electrochemiluminescent immunoassay system of Elecsys 2010]. AB - Performance of the electrochemiluminescent immunoassay system of Elecsys 2010 was evaluated by quantitative assay. The functional sensitivity of the TSH assay was 0.004 microU/ml. The extra-assay imprecision was RSD = 1.45%-4.65% in the range of TSH concentrations of 0.43-88.15 microU/ml. The intra-assay imprecision was RSD = 1.38%-3.14% in the range of TSH concentrations of 0.41-87.28 microU/ml. The recovery of TSH was in the range of 98.6%-104.4%. The correlation between theoretical values and obtained values of TSH was adequate(r = 0.999). We conclude that the Elecsys 2010 is provably satisfactory in sensitivity, precision, accuracy, reliability, practicability and rapidity. PMID- 12561375 TI - [Assessment of the role of TLR-4 in shear-stress-induced IL-8 gene transcription activation in vascular endothelial cells by gene mutation and gene transfection technology]. AB - This study is to evaluate the role of TLR-4 in lower laminar shear stress-induced interleukin-8 gene transcription activation in vascular endothelial cells by using gene mutation and transfection techniques. RT-PCR, Northern hybridization and immunocytochemical fluorescent staining showed that TLR-4 was expressed in human umbilical vein endothelial cells(HUVEC). When stimulated with 4.2 dyne/cm2 shear stress for 1 hour, an increase of TLR-4 mRNA expression was observed in HUVECs detected by RT-PCR and Northern hybridization. The intracellular domain deletion mutant TLR-4 cDNA (lacking the 155 COOH terminal amino acids of the wild type (TLR-4) and -102 -61 bp DNA sequence in 5'-flanking region of IL-8 gene (IL 8USCS) were isolated from endothelial cells by RT-PCR and PCR. These two DNA fragments were cloned into pcDNA3 and pEGFP1 respectively to construct TLR-4 dominant negative mutant pcDNA3-mTLR4 and IL-8 reporter gene pEGFP1-IL8USCS. ECV304 cells were transfected with the pEGFP1-IL8USCS or co-transfected with pEGFP1-IL8USCS and pcDNA3-mTLR4 by Dosper liposome transfectional reagent and selected by G418, then stimulated with 4.2 dyne/cm2 shear stress for 3 hours. Flow cytometric analysis showed that when exposed to 4.2 dyne/cm2 shear stress for 3 hours, there was a marked increase in the green fluorescent protein expression in pEGFP1-IL8USCS-transfected ECV304 cells. In contrast, there was almost no change in the green fluorescent protein expression in the cells co transfected with pEGFP1-IL8USCS and pcDNA3-mTLR4 after the stimulation, suggesting the TLR-4 mutant depressed TLR-4 signaling. These experiments suggest that the inflammatory TLR-4/NF-kappa B signaling pathway would probably be involved in flow shear stress-induced IL-8 gene expression in human vascular endothelial cells. PMID- 12561377 TI - [The advances in the application of amniotic membrane stroma in promoting tissue repair]. AB - Amniotic membrane is composed of amniotic epithelium, basement-membrane and stroma. Amniotic membrane is an easily obtained biomaterial and easily to be also processed, preserved and transported. However, its applicability will not be destroyed after it has been preserved for a long time(about one year). Thus it has been utilized widely in laboratory and clinical surgery. Generally, homologous amniotic membrane does not induce rejection after allotransplantation, and it is a bio-absorbable and degradable material. The purpose of this paper is to review the characteristics of amniotic membrane that makes it potentially useful in promoting tissue repair. PMID- 12561378 TI - [Structure, function and regulation of MMPs]. AB - The matrix metalloproteinases (MMPs) are a large family of proteolytic enzymes, which are involved in the degradation of many different components of the extracellular matrix. The MMPs have been classified into different groups including collagenases, gelatinases, stromelysins, membrane-type MMPs, etc. There is increasing evidence to indicate that MMPs play important roles in tumour invasion and metastasis. Moreover, MMPs not only play a direct role in tumour invasion by facilitating extracellular matrix degradation, but as a consequence they also have an important role in maintaining the tumour micro-environment and thus promoting tumour growth. In this paper, the structure, function and regulation of MMPs were reviewed. PMID- 12561379 TI - [Advances and prospective application of stem cell technique in stomatology]. AB - In this paper the background and advances of stem cell technique in stomatology were reviewed, especially the lately research of repair of maxillofacial defects with bone marrow stem cells, repair or reconstitution of teeth with dental pulp stem cells and repair of other tissues such as parotid with embryonic stem cell. Stem cell technique provides a new choice and extensive prospect of application for stomatology, therefore, deserves further research. PMID- 12561380 TI - [Progress in research and application of the tissue engineering blood vessels]. AB - Vascular tissue engineering is a novel approach by which an ideal vascualr graft constructed in vitro that will not be obstructed for a long time without immunological reaction after implantation. This article reviewed the definition of the tissue engineering blood vessel (TEBV), cellular resourses, the selection of biocompatible materials, the devising methods and the research achievements. Furthermore, it also discussed the current problems of TEBV and looked forward to future clinical application. PMID- 12561381 TI - [Research on signal sources for prosthetic limb control]. AB - Proper signal sources for prosthetic limb control are the premises in designing upper extremities with high effectiveness and multi-degree of freedom(DOF). With an overview of input sources adopted in prosthetic design, this paper reviews comprehensively on characters of signals available for arm control. PMID- 12561382 TI - [The study on the interleukin-8 (IL-8)]. AB - Interleukin-8 (IL-8), which is a member of C-X-C chemokine subfamily, is an important activator and chemoattractant for neutrophils and has been implicated in a variety of inflammatory diseases. Numerous reports show that various cells express IL-8 mRNA and produce IL-8 protein rapidly, including monocytes, T lymphocytes, neutrophils, fibroblasts, endothelial cells and epithelial cells. The human IL-8 gene has a length of 5191 bp and contains four exons separated by three introns. It maps to human chromosome 4q12-q21. The mRNA consists of a 101 bases 5' untranslated region, an open reading frame of 297 bases, and a long 3' untranslated region of 1.2 kb. The 5' flanking region of the IL-8 gene contains potential binding sites for several nuclear factors including activated protein-1 (AP-1), activated protein-2 (AP-2), nuclear factor-gene binding (NF-kappa B), nuclear factor-interleukin-6 (NF-IL-6, also calls CAAT/enhancer-binding proteins, C/EBP), IFN regulatory factor-1 (IRF-1), hepatocyte nuclear factor-1 (HNF-1), and so on. IL-8 gene expression is regulated initially at the level of gene transcription. The rapid induction of IL-8 gene expression is likely mediated by latent transcription factors that bind the IL-8 promoter. AP-1 and NF-IL-6 physically interact with NF-kappa B, and functional cooperativity among these factors appears to be critical for optimal IL-8 promoter activity in different cell types. The IL-8 receptor (IL-8R) is a dimeric glycoprotein consisting of a 59 KDa and a 67 KDa subunit. It has been given the name CDw128. It is expressed in many different cell types including those not responding to IL-8. The receptor density is approximately 20,000/cell in neutrophils, 1,040/cell in monocytes, and 300/cell in T-lymphocytes. The IL-8R is a member of the family of G-protein coupled receptors. There are at least two different IL-8 receptor types (CXCR1 and CXCR2). The activities of IL-8 are not species-specific. IL-8 affects the adhesion of neutrophils to the endothelium and induces the transendothelial migration of neutrophils. IL-8 also exhibits in vitro chemotactic activities against of T-lymphocytes and basophils. IL-8 gene expression can be regulated by fluid shear stress, which may play an important role in the genesis and development of both inflammation and arterosclerosis. PMID- 12561383 TI - [Gene therapy on spine fusion]. AB - Spine fusion stabilize adjacent vertebral segments by achieving bone union. The preferred method of spine fusion involves decortication of the host bed and transplantation of autologous bone graft from the iliac crest. Although autologous bone grafting is the gold standard, this procedure has significant complication. Gene therapy represents the new frontier of medical science that holds much promise for the improvement of spinal arthrodesis. Studies have shown the efficacy in using liposome-mediated and adenovirus-mediated gene transfer of bone morphogenetic protein (BMP) and related gene in animal models. Several studies in which gene transfer has been used specifically to enchance spine fusion in animal models are reviewed. Current main areas of research, including the elucidation of gene expression profiles during bone formation and the development of new gene transfer vehicles, promises the development of clinically applicable techniques in the near future. PMID- 12561384 TI - [Radioprotective effects on head and neck tumors of amifostine--a broad-spectrum cytoprotection]. AB - Differentiated thyroid cancer can be effectively treated with high-dose 131I and the other head and neck cancer can also be effectively treated with extra radiotherapy, but these treatments often result in a reduction in salivary gland function, causing xerostomia. Collectively, these effects can lead to severe secondary complications, including difficulty in speaking and swallowing, decreasing appetite even affecting nutrition and sleep. Amifostine, an analog of cysteamine, is a phosphorlyated aminothiol prodrug and its active metabolite, WR 1065 etc, can selectively protect normal tissues from the cytotoxic effects of drugs and/or radiation while preserve antitumor effects. Many studies have demonstrated that amifostine protects normal tissues from both acute and late extra-radiation damage without protecting the tumor. It has been approved by FDA to be used for protecting the salivary gland from xerostomia caused by radiotherapy. It has also show protecting effects on intra-radiotherapy, but there are many problems waiting for study. PMID- 12561385 TI - [Time-resolved fluorescence anisotropy measurement and its application in the research of biomacromolecule]. AB - In this symposium, we reviewed the principle and development of time-resolved fluorescence anisotropy measurement. Its method of measurement, characteristics and applications in the research of biomacromolecule, configuration and molecular structure have been discussed. Its potential applications are also illustrated. PMID- 12561386 TI - IRS ruling makes patient power a reality for all employers. AB - In June, 2002, the Internal Revenue Service released guidance on the use of Health Reimbursement Arrangements (HRAs) by employers. This action added another important tool to the effort to restore patient power to a financing system that has become ever more dysfunctional over the years. HRAs are far more flexible in design than earlier efforts such as Medical Savings Accounts or Flexible Spending Accounts. Ironically, because of the IRS, innovation is likely to continue in the area of employee benefits. PMID- 12561387 TI - Consumer driven healthcare and the birth of health reimbursement arrangements. AB - Consumers are quickly becoming more involved in the decisionmaking process as consumer-driven healthcare plans are surfacing across the country. When benefit programs are designed properly and when employees are properly informed, they can make wise decisions about healthcare that fits their needs and can help them save money. The process of engaging the plan member has come to be called consumer driven or consumer centric healthcare. The strategy of redefining responsibilities and costs between employer and plan member is generically referred to as a defined contribution strategy. Embedded in these efforts are choice, flexibility, and the belief that plan members can play an active part in managing costs when they are informed and empowered. Communication, education, and the use of Web-enabled technology are critical elements of this process. PMID- 12561388 TI - Case management redesign in a managed care system: one company's experience. AB - Strategies for redesign are necessary in successful managed care organizations as the member population grows and changes. This article presents a redesign project initiated by a managed care plan located in the Southwest region of the United States, where the rural frontier blends with metropolitan cities in a culturally diverse population. The focus of this project is the redesign of the Health Services department in which case management resides. PMID- 12561389 TI - Solving the claims conundrum. AB - Healthcare providers are being pushed to provide better service, yet maintain costs; and one of those costs is claims processing. There are many hurdles providers must jump over in the transition from capitation to "fee for service" billing structures, as well as potential issues surrounding new claim payment timeliness legislation. This article presents some possible solutions for making the claims process more efficient. PMID- 12561390 TI - Assessing managed care market variation in reports of coding accuracy. AB - The implementation of larger, faster and more comprehensive databases in healthcare delivery settings is an emerging outgrowth of evidence-based medicine. This study seeks to assess, at a national level, the degree of uniformity across markets in utilization and management of coded medical information. Implications for managers and policymakers, related to comparative managed care data benchmarks, are reviewed. PMID- 12561391 TI - Specialty pharmaceuticals: developing a management plan. AB - This is the first in a series of articles that address the complex issues associated with specialty pharmaceuticals in the development of a successful specialty pharmaceutical program, a critical component of managing this high-cost and highly fragmented sector. This article focuses on how to define specialty pharmaceuticals. Other articles in this series will explore such topics as the mechanics of developing and managing a specialty pharmaceutical program, how and when to establish clinical protocols and authorizations, the importance of data management, and the benefits from automated processes. PMID- 12561392 TI - Health plan responses to managed care regulation. AB - Little effort has been made to understand operational responses of health plans to the cascade of regulation imposed on them in the past decade. Using data from the Community Tracking Study, we cast some light on this area and illustrate how regulatory initiatives have produced both intended and unintended consequences. The findings both confirm speculation about regulatory effects and reveal some surprising and troubling developments. PMID- 12561394 TI - Can your members understand their health plan? PMID- 12561393 TI - Information therapy: prescribing the right information to the right person at the right time. PMID- 12561395 TI - Regulatory oversight of case management services and other medical management functions. PMID- 12561396 TI - Tiered benefit HMO plans: the next generation of managed care plans? PMID- 12561397 TI - Cannabis use in South Africa. PMID- 12561398 TI - Novel bicycle saddle--readers taken for a ride? PMID- 12561399 TI - The anaesthetist and the defibrillator--an urgent plea. PMID- 12561400 TI - Nevirapine all set to succeed in Gauteng. PMID- 12561401 TI - MCC slow on life-saving child vaccine. PMID- 12561402 TI - GP's vision kick-starts palliative training. PMID- 12561403 TI - Limb-saving prosthetics get cheaper. PMID- 12561404 TI - Gunderson links medicine and religion. Interview by Chris Bateman. PMID- 12561405 TI - A brief history of Pretoria Academic Hospital. PMID- 12561406 TI - Completeness of data on daily patient encounter forms at Mankweng Hospital. PMID- 12561407 TI - Incidence and outcome of caesarean section in the private sector--3-year experience at Pretoria Gynaecological Hospital. PMID- 12561408 TI - My suprapubic catheter. PMID- 12561409 TI - Kava kava may cause irreversible liver damage. PMID- 12561410 TI - The Pretoria academic complex--embracing the future. PMID- 12561411 TI - Fibrinogen Pretoria--a clinical and biochemical study of a Pretoria family with fibrinogen dysfunction resulting from a gamma chain defect. PMID- 12561412 TI - Conversion disorder and calcium homeostasis. PMID- 12561413 TI - Vascular disease in HIV/AIDS patients. AB - OBJECTIVES: An ongoing prospective clinical survey to determine the spectrum of vascular disease in HIV/AIDS patients and the risk factors affecting clinical outcome in order to formulate a management protocol for future use. METHODS: Comprehensive screening for risk factors for vascular disease as well as HIV/AIDS related conditions. Disease pattern and presentation are noted and patients treated accordingly. Vascular emergencies are managed regardless of HIV status because this information is usually not available at the time of presentation. Elective management is based on immune status and risk stratification. RESULTS: 42 patients tested positive for HIV. The majority of patients presented with occlusive disease (57%), followed by aneurysms (21%) and vascular trauma (19%). A variety of vascular surgical procedures were performed on 36 patients. There was no surgical mortality and 10 patients developed complications, including 2 amputations and 7 cases of minor wound sepsis. The 3 patients who received preoperative antiretroviral therapy showed a marked reduction in viral count and a significant improvement in CD4 T-cell count. CONCLUSION: Surgery can be safe and effective in HIV-positive patients provided the necessary precautions are taken to reduce surgical morbidity. PMID- 12561414 TI - Screening for childhood anaemia using copper sulphate densitometry. AB - OBJECTIVE: To evaluate copper sulphate densitometry to screen for childhood anaemia in a primary care setting, with a view to identifying children requiring definitive diagnostic testing and treatment. DESIGN: A cross-sectional screening study. Results of densitometry with a copper sulphate solution of specific gravity (SG) 1.048, corresponding to a haemoglobin (Hb) concentration of 10 g/dl, were compared with laboratory Hb determination. SETTING: Outpatient department of Pretoria Academic Hospital (73 children) and a local creche (27 children). SUBJECTS: One hundred consecutive children, aged between 6 months and 6 years, with informed written consent by parents. OUTCOME MEASURE(S): Accuracy of copper sulphate densitometry in screening for Hb concentration below 10 g/dl in terms of sensitivity, specificity, positive and negative predictive values, as well as likelihood ratio. RESULTS: The prevalence of anaemia (Hb < 10 g/dl) was 17% (95% confidence interval (CI) 10.2; 25.8). Copper sulphate densitometry had a sensitivity of 88.2% (95% CI 62.3; 97.9), a specificity of 89.2% (95% CI 79.9; 94.6), a positive predictive value of 62.5% (95% CI 40.8; 80.5) and a negative predictive value of 97.4% (95% CI 90.0; 99.5) in screening for anaemia. The likelihood ratio of a positive screening test was 8.17. CONCLUSIONS: Copper sulphate densitometry was accurate in screening for childhood anaemia. PMID- 12561415 TI - Sacrocolpopexy--a report on 262 consecutive operations. AB - OBJECTIVES: This report analyses the outcome and complications of 262 consecutive sacrocolpopexy procedures for the repair of vaginal vault prolapse and enterocele. METHODS: From March 1994 to February 2001, 262 patients underwent surgical repair using a standardised retroperitoneal technique. Initially dura mater strips were used and from the 19th patient onwards, Gore-tex soft tissue patch was used to suspend the vaginal apex to the anterior sacral ligament. Halban-type occluding sutures were placed in the pouch of Douglas. All patients were followed up and the minimum duration of follow-up was 16 months. RESULTS: Vaginal vault prolapse was successfully managed in 259 of 262 patients giving a success rate of 98.8%. In addition, 4 patients had a repeat enterocele that required surgical repair. The overall surgical complication rate was low. Erosion of the patch through the vaginal vault occurred in 10 patients, necessitating removal of the patch. Prolapse did not recur in any of these patients. CONCLUSION: Abdominal sacrocolpopexy is a very successful and safe surgical management of vaginal vault prolapse. PMID- 12561416 TI - Incidence of retinopathy of prematurity in very-low-birth-weight infants born at Kalafong Hospital, Pretoria. AB - INTRODUCTION: Retinopathy of prematurity (ROP) is a complication of prematurity, is diagnosed by ophthalmological screening of infants at risk (birth weight < or = 1,500 g), and may lead to blindness. The incidence of ROP is under-reported in developing countries, including South Africa. Published data from the USA (CRYO ROP) show that black infants have a lower incidence of threshold ROP than their white counterparts (3.2% v. 7.4%). Preliminary results of a screening programme initiated at Kalafong Hospital in 1999 are reported. AIM: To determine the incidence of ROP in infants with a birth weight of < or = 1,500 g born at Kalafong Hospital. PATIENTS AND METHODS: Consecutive infants were enrolled at birth and screened for ROP 4-6 weeks later by indirect ophthalmoscopy. Repeat examinations were performed until vascularisation was complete or until the infant reached a postconceptional age of 40 weeks. Infants with stage 3 ROP who developed threshold disease were treated with cryotherapy or laser therapy. RESULTS: One hundred and forty-five infants were enrolled over 10 months (15 February 1999-25 December 1999); of these 94 were screened. Of the remaining 51 infants, 24 died before screening and 27 were discharged before screening and were lost to follow-up. ROP was diagnosed in 23 of the 94 infants screened (24.5%). Stage 1 and 2 ROP occurred in 17 of the infants screened (18.1%) and stage 3 ROP in 6 (6.4%), of whom 4 (median birth weight 995 g, range 900-1,450 g) developed threshold ROP and were treated. CONCLUSIONS: The incidence of ROP in black very-low-birth-weight infants born at Kalafong Hospital is 24.5%. The incidence of threshold ROP is 4.3% (3.2% in infants < or = 1,250 g) and correlates with published data from the USA. Infants with a birth weight < or = 1,500 g should receive ophthalmological screening to diagnose stage 3 ROP timeously. PMID- 12561417 TI - Regulation of calcium homeostasis in activated human neutrophils--potential targets for anti-inflammatory therapeutic strategies. AB - OBJECTIVES: The objectives of the current study were to: (i) present an integrated model for the restoration of calcium homeostasis in activated human neutrophils based on current knowledge and recent research; and (ii) identify potential targets for the modulation of calcium fluxes in activated neutrophils based on this model and to investigate the effects of intracellular probes which target key processes involved in calcium homeostasis and pro-inflammatory activity in these cells. DESIGN AND SETTING: Laboratory-based experimental research using purified human neutrophils from healthy, adult human volunteers. OUTCOME MEASURES: Calcium metabolism and pro-inflammatory activity of neutrophils. RESULTS: Modulation of calcium fluxes in activated human neutrophils can be achieved by cAMP-dependent upregulation of the activity of the endomembrane Ca(2+)-ATPase which resequesters cytosolic Ca2+. Formoterol, a long acting beta 2-agonist, elevates intracellular cAMP levels, accelerates Ca2+ restoration in activated neutrophils and downregulates the pro-inflammatory responses of these cells. Alterations in the membrane potential of activated neutrophils may play a role in regulating calcium reuptake into the cells as attenuation of the membrane depolarisation response is associated with accelerated calcium influx. CONCLUSIONS: Modulation of the activity of the endomembrane Ca(2+)-ATPase in human neutrophils represents an important target for anti-inflammatory PMID- 12561418 TI - C20-diterpenoid alkaloids. PMID- 12561419 TI - The IBOGA alkaloids and their role as precursors of anti-neoplastic bisindole Catharanthus alkaloids. PMID- 12561420 TI - OR automation systems. AB - An operating room (OR) automation system is a combination of hardware and software designed to address efficiency issues in the OR by controling multiple devices via a common interface. Systems range from the relatively basic--allowing control of a few devices within a single OR--to advanced designs that are capable of not only controlling a wide range of devices within the OR but also exchanging information with remote locations. PMID- 12561421 TI - Intensive care ventilators. AB - Intensive care ventilators provide temporary ventilatory support to critically ill patients who cannot breathe on their own or who require assistance to maintain adequate ventilation. These ventilators offer a wide range of capabilities and can meet the needs of many types of patients. We last evaluated intensive care ventilators for adult use in our July-August 2000 issue. In the current Evaluation, we revisit the ratings of nine previously evaluated ventilators and describe our findings for one newly evaluated unit, the Siemens Servo-i. As ventilator technology has evolved, so have our ratings. Several units have added new features and capabilities since we last examined them. As a result, we have rated them higher in this study. Conversely, a few models have been given lower ratings because they haven't kept up with the advances in the technology; although they perform adequately, we can no longer recommend their purchase because better choices are available. PMID- 12561422 TI - Medical errors and patient safety. A report on ECRI's latest audio conference. PMID- 12561423 TI - Erratic control knobs affect Siemens Servo 300 and 300A ventilators. PMID- 12561424 TI - Rupture of Dyonics batteries creates risk of injury. PMID- 12561425 TI - Rainout puts ventilator-dependent patients at risk. PMID- 12561426 TI - [Identification of down-regulated expressed sequence tag at chromosome 3p21 in nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: It was reported that a locus of high frequency loss of heterozygosity was found at 3p14-25 of nasopharyngeal carcinoma (NPC) cells. This study was designed to identify the expressed sequence tags (ESTs) associated with NPC at chromosome 3p21 in order to clone new candidate NPC-associated genes at the locus. METHODS: The expression of relative ESTs at 3p21 was determined in nasopharyngeal carcinoma tissues and normal nasopharyngeal epithelia using ESTs homology analysis with bioinformatics in computer network combined with reverse transcription-polymerase chain reaction (RT-PCR). The expression of EST was detected in other normal tissues and tumor cell lines using Northern blot analysis. RESULTS: Comparing with expression of normal nasopharyngeal epithelia, EST (N31985) was down-regulated in 60.00% (3/5) nasopharyngeal carcinoma cell lines and 47.06% (16/34) nasopharyngeal carcinoma tissues (P < 0.05). CONCLUSION: N31985 is down-regulated in NPC, suggesting it may play a role in NPC carcinogenesis. PMID- 12561427 TI - [Abrogation of radiation-induced G2 arrest and radiosensitization by 7 hydroxystaurosporine (UCN-01) in human nasopharyngeal carcinoma cell line]. AB - BACKGROUND & OBJECTIVE: To enhance the radiosensitivity of cancer cell is one of the most important way to improve the effect of radiotherapy. This study was designed to investigate the radiosensitization of 7-hydroxystaurosporine(UCN-01) by its abrogation of radiation-induced G2 arrest. METHODS: Flow cytometry was used to observe the effect of irradiation and UCN-01 on cell cycle of human nasopharyngeal carcinoma cells (CNE-1) with mutated p53. The effect on radiosensitivity was determined by clonogenic assay and was quantified by calculating the sensitive enhancement ratio (SER). RESULTS: Irradiation resulted in G2 arrest in a dose-dependent manner. The proportion of cells at G2 phase, comparing to the control group (18.4%), was increased to 43.6%, 77.4% and 86.4% after 12 hours with irradiation of 2, 4 and 6 Gy, respectively. Radiation-induced G2 arrest was abrogated by UCN-01 in a concentration-dependent manner. UCN-01 at the concentrations of 50, 100, 200 and 400 nmol/L decreased the proportion of cells in G2 phase from 63.5% in the control group to 28.5%, 25.0%, 16.1% and 13.7%, respectively. UCN-01 resulted in a radiosensitization in CNE-1 cells. The SERs were 2.60 and 3.09, for 100 nmol/L and 200 nmol/L of UCN-01, respectively. CONCLUSION: Radiosensitization of UCN-01 in CNE-1 cells characterized by mutated p53 is associated with its abrogation of radiation-induced G2 arrest. PMID- 12561428 TI - [Apoptotic effect of bcl-XL antisense oligodeoxynucleotide mediated by lipofectin on cell strain CNE-2Z]. AB - BACKGROUND & OBJECTIVE: Recent studies have shown that overexpression of bcl-XL was detected in human nasopharyngeal carcinoma (NPC) cell strain CNE-2Z, suggesting it may play a pivotal role in tumorigenesis of NPC. The current study was designed to explore the effect of bcl-XL antisense oligodeoxynucleotide (ASODN) on CNE-2Z. METHODS: A 20-mer gapmer ASODN with a full phosphorothioate backbone targeting a sequence unique of the bcl-XL coding region was artificially synthesized. Bcl-XL ASODN was transfected into CNE-2Z cells through lipofectin. The survival rate was assessed by MTT assay and internucleosomal fragmentation of genomic DNA was detected by agarose gel electrophoresis. Apoptotic changes after treatment with ASODN were observed by fluorescence microscopy and flow cytometry. RESULTS: MTT assay showed that the proliferation of CNE-2Z cells decreased significantly after treatment with ASODN/Lip as compared with control (P < 0.01). ASODN/Lip reduced the proliferation of CNE-2Z in a dose-dependent manner. After treatment with ASODN/Lip for 36 hours, most cells stained with Hoechst 33258/Pl exhibited apoptotic cell morphology such as cell shrinkage, nuclear condensation, and nuclear fragmentation under fluorescence microscope; a apoptotic peak appeared on flow cytometry; a ladder-like pattern of DNA fragmentation appeared on agarose gel electrophoresis. CONCLUSION: ASODN can inhibit proliferation of CNE-2Z cells and induce apoptosis of CNE-2Z cells. The results suggest that bcl XL is a promising target for gene therapy of NPC. PMID- 12561429 TI - [Mutation and expression of SEMA3B and SEMA3F gene in nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: Though the molecular etiology of nasopharyngeal carcinoma(NPC) is currently unknown, evidence from both loss of heterozygosity analysis and functional studies suggested that there are NPC-associated tumor suppressor genes(TSGs) residing in chromosome 3p21.3. Recently, two members of semaphorin family, SEMA3B and SEMA3F gene, located at 3p21.3, were characterized as TSGs. Studies showed that SEMA3B and SEMA3F are capable of suppressing the growth of tumor cells and inducing apoptosis. Loss of SEMA3B mRNA expression or aberrant SEMA3F cellular localization were found in lung cancers. In order to investigate the involvement of SEMA3B and SEMA3F in NPC, the authors examined both mutation and expression of these two genes in NPC. METHODS: The entire coding regions, the splice donor/acceptor sites, and partial regulatory regions of SEMA3B and SEMA3F gene were screened for mutations by PCR-sequencing in 21 primary NPC tumors and 2 NPC cell lines(CNE2 and SUNE1). The mRNA expression levels were determined by semi-quantitative RT-PCR analysis. RESULTS: No somatic mutation was found in either SEMA3B or SEMA3F gene. However, two missense polymorphisms including Thr415Ile and lle242Met were found in SEMA3B in NPC. For the Thr415Ile polymorphism, the Ile allele type which leads to SEMA3B function defects was predominant in NPC with the allele frequency of 64% (27/42). SEMA3B mRNA was expressed in all 6 non-neoplastic nasopharyngeal epithelia, but was absent or down-regulated in 76% (16/21) of primary NPC tumors. No significant difference of SEMA3B expression was observed between NPC and noncancerous controls. CONCLUSION: High frequency of SEMA3B expression alterations suggests that the inactivation of this gene was strongly associated with NPC. SEMA3B may be a tumor suppressor on 3p21.3 involved in NPC. PMID- 12561430 TI - [Arsenic trioxide induces differentiation of human nasopharyngeal carcinoma in BALB/C nude mice xenograft model]. AB - BACKGROUND & OBJECTIVE: There is evidence that arsenic trioxide (AS2O3) induce differentiation of leukemia cells; however, little is known about its effect on solid tumors. The aim of this study was to investigate whether AS2O3 can induce cell differentiation and its association with growth inhibition in human nasopharyngeal carcinoma using BALB/C nude mice xenograft model. METHODS: Poorly differentiated human nasopharyngeal squamous carcinoma cells from CSNE-1 cell strain were transplanted subcutaneously to BALB/C nude mice to produce tumors. AS2O3 at a dose of 5 mg.(kg.d)-1 was given intraperitoneally for 10 consecutive days, and then 3 times a week for the following 3 weeks. The xenograft tumor growth in mice was observed after drug administration. The morphological changes of the tumors were examined under light and electron microscopy. Proliferating cell nuclear antigen (PCNA) expression was determined by immunohistochemistry. RESULTS: AS2O3 at dose of 5 mg.(kg.d)-1 significantly inhibited the tumor growth in vivo, with a inhibitory rate of 75.4%. Remarkable cell differentiation induced by AS2O3 was observed under light microscope and transmission electron microscope, which was characterized by keratinization of tumor cells, decreased nuclear/cytoplasm ratio, increased cytoplasmic organelles and rich tonofibrils in cytoplasm. Desmosomes and micro-processes were much more frequently observed in tumors treated with AS2O3. Significantly decreased PCNA expression was observed in AS2O3-treated tumor cells. The PCNA-positive cell index (PI) was 53.6 +/- 7.0% in AS2O3-treated mice, and 95.2 +/- 5.0% in control, respectively (P < 0.001). CONCLUSION: The growth of human nasopharyngeal carcinoma xenograft in BALB/C nude mice can be significantly inhibited by AS2O3, which might be related to the cell differentiation induced by AS2O3. PMID- 12561431 TI - [Influence of selenium-rich rice on transformation of umbilical blood B lymphocytes by Epstein-Barr virus and Epstein-Barr virus early antigen expression]. AB - BACKGROUND & OBJECTIVE: Selenium (Se), an antioxidant, is an essential trace element to human body. It can be used as an anti-aging agent and a tumor cell proliferation inhibitor. To further investigate the effect of selenium in cancer prevention, the authors observed the influence of Se-rich rice extract on the transformation of umbilical blood B lymphocytes stimulated by Epstein-Barr virus (EBV) and expression of EBV early antigen(EBV-EA) in Raji cells. METHODS: (1) Se rich rice and general rice extract (dilution of 1:4 or 1:8) were added to mixture of EBV, and then umbilical blood mononuclear cells were added. Lymphoblasts transformation test was then performed. The inhibition rate of B lymphocytes transformation was calculated. (2) Raji cells stimulated by butyrate and croton oil were incubated with Se-rich rice extract. The EBV-EA positive expression rate and the inhibition rate were counted using indirect immunological flurescence method. RESULTS: The transformation of umbilical blood B lymphocytes stimulated by EBV was significantly inhibited by Se-rich rice extract at a concentration of 0.11 g/ml (1:8 diluted). The inhibition rate was 83.4% (P < 0.01), which was significantly higher than that of the control rice (63.1%) (P < 0.05). Se-rich rice extract showed significant inhibition on EBV-EA in Raji cells. As the extract concentration was at 0.016 microgram/ml, 0.078 g/ml, and 0.388 microgram/ml, the inhibition rates of EA were 2.85%, 12.88%, and 20.75%, respectively. CONCLUSION: The transformation of umbilical blood B lymphocytes stimulated by EB virus and expression of EBV-EA in Raji cells may be significantly inhibited by Se-rich rice extract, suggesting that Se-rich rice can be used for preventing nasopharyngeal carcinoma. PMID- 12561432 TI - [Influences of PAX3-FKHR transfection on expression of MRF4 and differentiation of cultured human rhabdomyosarcoma RD cells]. AB - BACKGROUND & OBJECTIVE: There were a few reports about the role of PAX3-FKHR fusion gene in the oncogenesis of alveolar rhabdomyosarcoma (ARMS); however, its effect on muscle-producing factor (MPF) is not clear. This study was designed to observe the effects of fusion gene PAX3-FKHR transfection on expression of MRF4 and differentiation of cultured human rhabdomyosarcoma RD cells. METHODS: To construct an eukaryotic expression vector of PAX3-FKHR gene; the plasmids of MRF4 cDNA and PAX3-FKHR cDNA were transfected into cultured RD cells with lipofectin methods. The changes of cells growth, morphologic and the expression of MHC and alpha-actin were observed. RESULTS: There was no significant changes in RD cells after transfection with PAX3-FKHR gene in growth rate, growth manner and the expression of MHC and alpha-actin. In MRF4 expressing RD cells after transfected with PAX3-FKHR, although the expression of MRF4 was not changed, morphologic appearance showed a low differentiation; expression of MHC and alpha-actin significantly decreased. CONCLUSION: There is no obvious influence on biological character and differentiation in RD cells with transfection of PAX3-FKHR gene; PAX3-FKHR does not inhibit the expression of MRF4, but can reverse the effect of MRF4 on promoting the differentiation of RD cell. PMID- 12561433 TI - [Quantitative analysis of p53 and related genes mRNA in rat hepatocarcinogenesis induced by 3'-Me-DAB]. AB - BACKGROUND & OBJECTIVE: p53 gene mutations and abnormal expression of p53 in hepatocarcinoma have been reported, but alteration in mRNA level is not yet understood. In order to find out the alteration in mRNA levels of p53, glutathione S-transferase P (GST-P), alpha-fetoprotein (AFP), and albumin in genesis, developing, and prognosis of hepatocarcinoma, quantitative analysis of mRNA levels of p53, GST-P, AFP, and albumin in prehepatocarcinoma and hepatocarcinoma foci was performed. METHODS: During hepatocarcinogenesis of F344 rat induced by 3'-methyl-4-dimethylamino-azobenzene (3'-Me-DAB), the levels of these mRNAs were quantitatively analyzed by LightCycler V3 System real-time RT PCR after capturing accurately micro-foci in prehepatocarcinoma and hepato carcinoma of rats with laser capture microdissection (LCM). RESULTS: At the 6th, 12th and 24th experiment weeks, the p53 mRNA levels in all of prehepatocarcinoma foci were markedly higher than those in the adjacent normal tissues (all of P < or = 0.001), and gradually decreased from the 6th week to the 24th week (P < 0.01). The content of p53 mRNA in hepatocarcinoma foci was higher than that in normal tissue (P = 0.028 and 0.013), but lower than that in prehepatocarcinoma foci. At 24th weeks, the sections of livers exhibited intensive immunostaining of p53 protein in prehepatocarcinoma and hepatocarcinoma foci. At any time-point of experiment, GST-P mRNA levels in prehepatocarcinoma foci were significantly higher than those in the adjacent normal liver tissue and hepatocarcinoma foci (all of P < 0.001). The concentration of AFP mRNA was the highest (P < 0.001) and that of albumin mRNA was the lowest (P < 0.01) in hepatocarcinoma foci as compared with adjacent normal tissue and prehepatocarcinoma foci. The GST-P protein and AFP protein were expressed strongly in prehepatocarcinoma and hepatocarcinoma foci, respectively. CONCLUSION: GST-P and AFP mRNA overexpressed in prehepatocarcinoma and hepatocarcinoma foci respectively will be profitable markers for diagnosis during hepatocarcinogenesis. The p53 mRNA highly expressed at early stage of prehepatocarcinoma and hepatocarcinoma, but the increasing concentration of p53 protein was found at later stage. PMID- 12561434 TI - [Expression of phosphatase and tensin homology deleted on chromosome ten(PTEN) and p53 protein and their significance in human hepatocellular carcinomas]. AB - BACKGROUND & OBJECTIVE: Tumor suppressor gene phosphatase and tensin homology deleted on chromosome ten (PTEN) was first reported in 1997 and has become a hot spot shortly after discovery. Mutations of PTEN gene in human hepatocellular carcinomas (HCCs) have been examined by several groups abroad, while little is known about expression of PTEN in HCCs. It has been believed that p53 mutation plays a role in the oncogenesis of HCCs. However, divergence exist as regards the relationship between p53 mutation and some biological behaviors of HCCs. The current study was designed to investigate the correlations of expression of PTEN and mutant p53 with carcinogenesis and tumor progression of HCCs. METHODS: Sixty two paraffin-embedded hepatocellular carcinoma samples were composed and all the tissue sections included both HCC and surrounding non-tumor liver tissues. Expression of PTEN and mutant p53 was examined using SP immunohistochemical technique. RESULTS: All sixty-two samples showed decidedly positive staining of PTEN in surrounding non-tumor liver tissues (plasma). While, only twenty-nine tumors were detected positive staining of PTEN (46.8%). The expression level of PTEN in HCC samples without invasiveness or with complete capsule was higher than that in HCC samples with invasiveness or without complete capsule. No positive staining of p53 was detected in surrounding non-tumor liver tissues for all sixty two samples. However, twenty-one tumors showed positive staining of p53 (nucleon). The expression level of p53 protein was not related with invasion or differentiation of HCCs. CONCLUSION: Loss of PTEN expression and mutation of p53 may play a role in occurrence of HCCs. Loss of PTEN expression may be pertinent to deterioration of HCCs. PMID- 12561435 TI - [Influence of p16 gene on the biological behavior of human hepatocellular carcinoma]. AB - BACKGROUND & OBJECTIVE: Inactivation of the tumor suppressor gene p16INK4a is one of the most common genetic alterations in human hepatocellular carcinomas (HCC), making it an ideal target gene for treatment of HCC. The objective of this study was to investigate the influence of wild p16 gene on the biological behavior of HCC. METHODS: HCC cell strains SNU-449 (loss of p16 protein expression) and HepG2.2. 15 (positive p16 protein expression) were respectively infected by a retrovirus expression vector of p16 gene (pcLXSN-p16). The stable p16 protein expression cell strains were selected. The biological behaviors of the p16 gene transfected HCC cells were observed. RESULTS: SNU-449 with negative p16 protein expression demonstrated that pcLXSN-p16 treatment significantly inhibited cell growth (the amount of cells at G0-G1 phase increased). However, there was no treatment effect when pcLXSN-p16 was transfected in HepG2.2. 15 which has positive p16 protein expression. Subsequent study in a nude mouse model demonstrated that the p16 gene transfected SNU-449 had a lower succeeding rate of first time establishment of tumors and grew more slowly in the nude mice as compared with non-transfected SNU-449. Moreover, the nude mice inoculated with transfected SNU-449 had a longer survival time than those inoculated with non transfected SNU-449. CONCLUSION: The transfer of wild p16 gene can inhibit the proliferation and reduce the invasion ability of HCC cells with p16 negative expression, but can not affect the HCC cells with p16 positive expression. PMID- 12561436 TI - [Mutations of fragile histidine triad gene in Peutz-Jeghers syndrome and canceration]. AB - BACKGROUND & OBJECTIVE: Peutz-Jeghers syndrome (PJS) is an autosomal dominantly inherited disease. Fragile histidine triad (FHIT) gene is an important tumor suppressor gene at the fragile sites region of 3p14. The authors' previous study suggested that PJS patients might have a susceptible gene at the region of 3p14.2. This study was designed to reveal the relationship between the variant of FHIT gene in PJS and its canceration. METHOD: Mutations of FHIT gene in 15 PJS patients and 20 unaffected members in 6 PJS families were determined using denaturing high-performance liquid chromatography (DHPLC), polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) and DNA sequencing techniques. RESULTS: A non-sense mutation and a frame-shift mutation were identified at codon 54(GAA to TAA) (exon 6) which led to the change of the amino acid from glutamic acid (Glu) to stop codon, and a guanine insertion at codon 62 in exon 6 resulting in a premature stop codon TGA at codon 111 in one PJS patient. A homozygous deletion and a synonymous mutation were detected in exon 8. The homozygous deletion of exon 8 in FHIT gene was found in two polyps tissues and two cancerous tissues. And in 3 sporadic cases, the patients and their mothers have the same bands of SSCP and the same elution profiles of DHPLC when exon 8 was amplified. The DNA sequencing result showed that a synonymous mutation (polymorphism) occurred at codon 98 [CAT (H)-->CAC (H)], this mutation resulted in no change of amino acid. In addition, one base substitute from A to G mutation at 5'end, +42 nucleotide in intron 6 of FHIT gene was detected in seven patients and two unaffected members. CONCLUSION: PJS patients have low frequency point mutation of FHIT gene and their cancerous tissues had homozygous deletions in FHIT gene. This study indicated that the mutations and deletions of FHIT gene in PJS may play a role in the development of PJS and their cancerations. PMID- 12561437 TI - [Modified intestinal in situ neobladder in functional reconstruction of lower urinary tract after radical cystectomy: report of 15 cases]. AB - BACKGROUND & OBJECTIVE: Quality of life (QOL) in the patients who are performed radical cystectomy is affected by the methods of urinary diversion. The patients with continent or non-continent urinary diversion could not void by themselves and their QOL was poor. However, the intestinal neobladder in situ could provide the patients with good voiding and improve QOL. Unfortunately, some degree of poor empty and incontinence would occur in these patients. The aim of this paper was to report the authors' experience of modified intestinal in situ neobladder in reconstructing the lower urinary tract after radical cystectomy. METHODS: Reconstruction of the lower urinary tract using modified sigmoid neobladder (in 12 patients) and modified ileal neobladder (in 3 patients) was carried out in 15 patients (male 14, female 1; age 33-68 years, mean 53 years) who underwent radical cystectomy for invasive bladder cancer. The patients were followed up for 3-30 months. Nine of them were followed up for more than 16 months. Clinical outcome of these patients including the function of the neobladder, urinary function, sexual status, renal function, serum electrolytes, and QOL was evaluated. RESULTS: All patients voided well by themselves and did not need self catheterization. Thirteen patients were continent in daytime and night-time. One patient was continent in daytime, but had mild incontinence at night. A female patient had moderate stress incontinence. The capacity of the neobladder was 240 640 ml and the residual volume was 0-250 ml. Renal function was normal in all patients. Serum electrolytes were normal in 14 patients. Metabolic acidosis and unilateral ureteral dilation was encountered in one patient respectively. Nine male patients restored sexual function and 13 patients came back to work. All patients felt satisfied with the function of the neobladder. CONCLUSION: Satisfactory urinary continence and voiding function was achieved with modified intestinal neobladder, which was believed an ideal procedure for lower urinary tract reconstruction after radical cystectomy. PMID- 12561438 TI - [Serum level and expression of vascular epithelial growth factor in ovarian epithelial carcinoma]. AB - BACKGROUND & OBJECTIVE: This study was designed to examine the level of serum vascular epithelial growth factor (VEGF) and VEGF expression in the patients with ovarian epithelial carcinoma (EOC) and to evaluate the relationship between VEGF and the clinicopathological factors. METHODS: Seventy-three patients with ovarian epithelial tumor were selected as study group, including 24 benign epithelial ovarian tumor (BET), 7 borderline epithelial ovarian tumor (BOT), 42 EOC. Twenty women without obvious gynecologic diseases were selected as serum control group (CON). VEGF expression in the epithelial ovarian tumor tissues from 73 patients were analyzed immunohistochemically. Serum VEGF in 71 patients and ascite VEGF in 10 patients were detected by enzyme-linked immunosorbent assay (ELISA) and 7 cases with EOC were continuously determined after operation. RESULTS: (1) Positive immunostaining of VEGF was observed in 86.3% of EOC, which was significantly higher than that of BOT (66.67%, P < 0.005) and BET (37.5%, P < 0.005). (2) There were significant differences in serum VEGF level among BET, BOT, and EOC group (P < 0.01). Serum VEGF levels before operation in patients with EOC were significantly higher than those with BOT, BET, and CON (P < 0.01). Serum VEGF levels in EOC patients at advanced stage (stage III or IV), low differentiation (G3) were apparently higher than those at early stage (stage I or II) (P < 0.05) and high differentiation (G1, G2) (P < 0.01), respectively. Serum VEGF levels decreased after the successful removal of tumor in ovarian cancer patients. The serum VEGF levels were re-elevated during relapse in three patients. CONCLUSION: (1) The elevated serum VEGF levels and increased VEGF expression in EOC were associated with its maligment behavior. (2) Serum VEGF could be used as a marker for monitoring the clinical course of the patients with epithelial ovarian carcinoma. PMID- 12561439 TI - [Prognostic significance of cellular DNA content and AgNOR count in ovarian cystadenoma]. AB - BACKGROUND & OBJECTIVE: It has been showed that cellular DNA content and silver staining nucleolar organizer region (AgNOR) count were significantly associated with the prognosis of some tumors; however, it is not yet clear whether these two parameters be helpful in predicting the prognosis of ovarian cystadenoma. This study was designed to evaluate the prognostic significance of these two parameters in ovarian cystadenoma. METHODS: Cellular DNA content and AgNOR count of the tumor cells for the samples from 42 patients with ovarian cystadenoma were measured by using computerized image analysis. The relationship between clinical as well as histomorphologic factors and the overall survival was evaluated by multivariable analysis. RESULTS: Univariate analysis showed that AgNOR count, DNA index, FIGO stage, degree of differentiation, presence of distant metastasis, residual tumor, lymph node metastasis, and age were the clinical variates with significant difference (P < 0.05). Multivariate analysis showed that DNA index > 1.4, FIGO stage, degree of differentiation, and residual tumor were prognostic predictors of ovarian cystadenoma. CONCLUSION: The DNA index may be a useful independent factor for predicting the diagnosis of ovarian cystadenoma. PMID- 12561440 TI - [Clinical and pathological related factors of occult cervical lymph node metastasis in squamous cell carcinoma of tongue]. AB - BACKGROUND & OBJECTIVE: Some clinical and histopathological features of squamous cell carcinoma of tongue (TSCC) were closely associated with occult cervical lymph node metastasis (OCLNM). This study was designed to investigate the correlation between occult cervical lymph node metastasis in squamous cell carcinoma of tongue and the clinicopathological indexes of primary tumor tissue. METHODS: Thirty-five TSCC patients with OCLNM treated in Cancer Center, Sun Yat sen University from 1988 to 1996 were enrolled. According to random principle, 35 TSCC patients without OCLNM at the same period were selected. Tumor deepness was measured and pathological parameters were assessed under microscope. RESULTS: The rates of OCLNM in group of infiltrative plus ulcerative type, group of infiltrative type, group of ulcerative type and group of exogenous type were 70.37%, 41.67%, 40.00%, and 27.27%, respectively; the rates of OCLNM in T1, T2, and T4 were 44.00%, 43.48%, and 62.64%, respectively. The rates of OCLNM in groups of tumor deepness < 4 mm, 4-7.9 mm, and > or = 8 mm were 14.29%, 61.54%, and 88.89%, respectively. The rates of OCLNM in groups of response of peritumoral lymphocyte +, ++ and +++ were 73.68%, 58.62%, and 18.18%, respectively. The rates of OCLNM in groups of with vascular invasion and without vascular invasion were 85.71% and 46.03%, respectively. The rates of OCLNM in groups of well, moderate, and poor differentiation were 52.63%, 42.31%, and 66.67%, respectively. The rates of OCLNM in groups of invasive patterns of type I, type II, type III, and type IV were 40.90%, 50.00%, 52.00%, and 80.00%, respectively. Univariate analysis indicated that OCLNM in TSCC was closely associated with gross appearance of primary tumor, tumor deepness, response degree of peritumoral lymphocyte and vascular invasion (P < 0.05), but was not associated with T stage, pathological differentiation, and invasion way (P > 0.05); Logistic multivariate analysis manifested that only tumor deepness and response degree of peritumoral lymphocyte significantly correlated with OCLNM in TSCC. CONCLUSION: There was significant correlation between gross appearance of primary tumor, tumor deepness, response degree of peritumoral lymphocyte and vascular invasion and OCLNM in TSCC. These parameters can be used to predict occult cervical lymph node metastasis in TSCC. PMID- 12561441 TI - [Application of microwave coagulation in treatment of cancer of oral cavity: summary of 96 cases]. AB - BACKGROUND & OBJECTIVE: There was few report about the protection of oral function of microwave coagulation in treatment of cancer of floor of the mouth. This study was designed to systematically clarify the principle, operation, treatment of the primary cancer and lymph nodes, efficiency, function of the mouth, complications and indications of the microwave coagulation in treatment of the cancer of the oral cavity. Based on this, the significance of the two special methods and prevention of bleeding during the cause were discussed. METHODS: Ninety-six cases with cancer of oral cavity who were treated with microwave coagulation were analyzed retrospectively. The survival rates and the recurrent rates of those with the coagulated tissue be resected or unresected, and of the positive margin group and the negative margin group were compared for analyzing the complications and prevention of them. RESULTS: The difference of the survival rates of those whose coagulated tissue pathologically positive and negative was insignificant (Log Rank = 0.70, P = 0.4033), and the differences of the recurrent rates of the primary site, secondary site and the uncertain site of them were also insignificant (chi 2 = 1.65, 0.837, 0.003; P = 0.684, 0.36, 0.959). The overall survival rate of the patients with negative margin was better than those of with positive margin (Log Rank = 6.08, P = 0.0136). However, the difference of the recurrent rates of them was insignificant (chi 2 = 0.327, P = 0.567). The total complication rate of 96 patients was 9.6% and all of them were acceptable. The complication rate of those with the coagulated tissue be resected was lower than that of the unresected (P = 0.013). CONCLUSION: Ligating the tongue artery, resecting the coagulated tissue, and packing the wound margin were effective methods to prevent the bleeding. The pathological result of the coagulated tissue could not be used to predict the prognosis. It is best to hold enough treatment range for microwave coagulation. Application of microwave coagulation in treatment of cancer of oral cavity has the advantage of less and slight complications. PMID- 12561442 TI - [Correlation between expression of p53 and prognosis of laryngeal squamous carcinoma]. AB - BACKGROUND & OBJECTIVE: p53 expression was a marker to predict the poor prognosis of laryngeal cancer. Detection of p53 in the pathological negative margin can be used to find more patients with poor prognosis. The aim of this study was to reveal the correlation between p53 expression and prognosis in primary cancer and surgical margin of laryngeal squamous cell carcinoma (SCC). METHODS: The clinical data of 67 laryngeal SCC with pathological negative margin were analyzed retrospectively. The expression of p53 protein in the margin and primary cancer was detected by immunohistochemistry. The survival rates and the recurrent rates were compared between p53 positive and negative group both in surgical margin and primary cancer. RESULTS: (1) The positive rates of p53 in surgical margin and primary cancer were 19.4% (13/67) and 50.7% (34/67), respectively. (2) The recurrent rate of positive margin group was higher than that of negative margin group (69.2% vs. 33.3%, P = 0.018). The survival rate of positive margin group was lower than that of negative margin group (24.62% vs. 75.69%, P = 0.0012). The recurrent rate of p53 positive primary cancer was higher than that of p53 negative primary cancer (64.7% vs. 15.2%, P = 0.0000). The survival rate of p53 positive primary cancer was lower than that of p53 negative primary cancer (42.86% vs. 90.86%, P = 0.0000). CONCLUSION: The prognosis of those with p53 protein positive in the pathological margin or the primary cancer were worse than that of the negative group. PMID- 12561443 TI - [Expression of tumor marker CDw75 and its clinical impact in gastric carcinoma]. AB - BACKGROUND & OBJECTIVE: Tumor-associated expression of sialylated glycoconjugates was found to be closely associated with aggressive behavior and increased metastatic potential of malignant cells. The CDw75 (cluster of differentiation antigen 75) epitope is a sialylated carbohydrate determinant generated by beta galactosyl alpha 2, 6-sialyltransferase. This study was designed to investigate the expression of CDw75 in gastric carcinoma and normal gastric mucosa, and the relationship between its expression and the infiltration, metastasis, and clinical stage of gastric cancer. METHODS: The expression of CDw75 antigen in 54 cases of gastric carcinoma and normal gastric mucosa was detected using immunohistochemical method. The percentage of CDw75 positive cells was counted through the computer analysis of picture. RESULTS: There was no expression of CDw75 antigen in normal gastric mucosa, while positive staining was detected in 26 (48.1%) of the 54 gastric cancer (P < 0.05). The expression of CDw75 antigen was significantly associated with invasive depth, TNM stage, and lymph node metastasis (P < 0.05 or P < 0.01). CONCLUSION: The expression of CDw75 in tumor tissue may be a new marker of the biological behavior of gastric carcinoma. PMID- 12561444 TI - [Expression of cyclin D1 and vascular endothelial growth factor(VEGF) in non small cell lung carcinoma and their association with the prognosis]. AB - BACKGROUND & OBJECTIVE: The occurrence and development of tumors are controled by oncogene, antioncogene and tumor metastasis-related gene. Cyclin D1 is the expressive product of CCND1(a kind of oncogen). Vascular endothelial growth factor (VEGF) regulates the growth of neoplastic angiogenesis, which plays a role in the invasion and metastasis of tumor. The objective of this study was to detect the expression of Cyclin D1 and VEGF in non-small cell lung cancer (NSCLC), and to explore their association with the prognosis of NSCLC. METHODS: Immunohistochemistry was used to detect the expression of Cyclin D1 and VEGF in pathological tissue sections of 55 cases of NSCLC and 10 cases of non-malignant tissue from lung lesions. The relationship between the expression of Cyclin D1 and VEGF in 55 NSCLC sections and the age, sex, smoke, size of tumor, histopathological type, differentiation, stage, lymph node metastasis and survival time were analyzed statistically. RESULTS: The expression rates of Cyclin D1 and VEGF in the 55 NSCLC tissue sections were 61.82% and 74.55%, respectively. In 10 cases of non-malignant tissue sections, cyclin D1 expression was not detected and VEGF expression (+/-) was only in 2 cases. The expression of Cyclin D1 and VEGF showed: (1) there was no significant difference among age, sex, histopathological type, stage, differentiation, tumor size and smoking level; (2) there were significant differences between the patients with and without lymph node metastasis; (3) there were significant differences of survival time between positive and negative expression groups. It meant Cyclin D1 and VEGF would be the poor prognostic factors. CONCLUSION: The expression of Cyclin D1 and VEGF occurred in more than 60% cases of NSCLC, which may play a role in the biologic behavior and prognosis of NSCLC. PMID- 12561446 TI - [Expression of androgen receptor protein in hormone refractory prostate cancer]. AB - BACKGROUND & OBJECTIVE: It was reported that androgen receptor (AR) gene amplification was found in hormone refractory prostate (HRPC) and it has been thought to be a possible molecular mechanism for the treatment failure. In this article, the expression of AR protein before and after hormone therapy was examined quantitatively in order to further clarify the relationship between AR expression and the genesis of HRPC. METHODS: AR protein content was determined using radioligand binding assay before and after hormone therapy in 28 patients with advanced prostate cancer. RESULTS: The average levels of AR protein before and after hormone therapy were 390.0 +/- 204.1 fmol/mg protein and 690.4 +/- 444.0 fmol/mg protein, respectively (P < 0.001). In 10 patients whose diseases recurred in 12 months after therapy, the mean levels of AR protein before and after therapy were 398.2 +/- 199.5 fmol/mg protein and 448.2 +/- 274.1 fmol/mg protein, respectively (P > 0.20). In other 18 patients with response duration more than 12 months, the mean levels of AR protein before and after therapy were 386.4 +/- 212.3 fmol/mg protein and 824.9 +/- 468.6 fmol/mg protein, respectively (P < 0.001). CONCLUSION: The elevation of AR protein level was most likely to occur in the prostatic tumors that had good response to hormone therapy and might be involved in the genesis of HRPC. PMID- 12561445 TI - [Cox regression analysis of the prognostic factors of unresectable esophageal carcinoma after stenting]. AB - BACKGROUND & OBJECTIVE: There are many factors affect the prognosis of the patients with unresectable esophageal carcinoma who underwent intubation, however, it is unclear which ones are main causes so far. This study was designed to analyze the prognostic factors of unresectable esophageal carcinoma after stenting in order to find the reasonable modalities of palliative therapy. METHODS: Consecutive 102 patients with unresectable esophageal carcinoma who were eligible for inclusion criteria were analyzed after stenting. Twelve factors including gender, age tumor site, tumor length, stricture degree in diameter, pathologic type, grade of cell differentiation, clinical tumor stage (T, N, M), pre-stenting therapy and post-stenting therapy (radiotherapy and/or chemotherapy) were used for Cox regression model analysis. The survival rate was calculated by life table. RESULTS: The technical sucess rate of stenting was 98.3%. There were significant improvement on the grade of swallowing function (P = 0.000) and quality of life (KPS scores, P = 0.000). The incidence rate of complications was 43.1% (44/102). The survival rates of 3, 6, 9, 12 months were 67.53%, 40.59%, 27.43%, and 18.65%, respectively. The results of Cox regression showed that invasion degree of primary tumor (T, P = 0.0410) and distal metastasis (M, P = 0.006) were the statistically significant prognostic factors. The odds ratio(ORT) was 1.750 (95% CI, 0.996-3.074) on T stage of primary carcinoma, ORM was 1.527 (95% CI, 1.126-2.069) on M stage. There was no statistical significance in the survival of the patients affected by radiotherapy and/or chemotherapy after intubation. CONCLUSION: T stage and M stage are the major prognostic factors affecting the survival of patients with unresectable esophageal carcinoma after stenting. There is no benefit for survival of patients treated with radiotherapy and/or chemotherapy after intubation. PMID- 12561447 TI - [Research advances on bladder cancer associated genes]. AB - Bladder cancer is the most common urinary tumors in China. Carcinogenesis of bladder is a multistep process. Accumulation of abnormal genotypes in a long period leads to malignant phenotypes. The genes associated with bladder carcinogenesis include oncogenes (such as H-ras, FGFR3, erbB2, CCND1, mdm2), tumor suppressor genes (such as INK4A/ARF, Rb, TP53, PTEN, TSC1, PTCH, DBCCR1), and DNA mismatch repair genes, etc. In this review, the authors discussed the recent research advances on the genes associated with bladder carcinoma. PMID- 12561449 TI - Greenhouse suits. Litigation becomes a tool against global warming. PMID- 12561448 TI - [Research advances on alpha-fetoprotein physiological function and clinical potential]. AB - Alpha-fetoprotein (AFP), a tumor-associated fetal protein, has been employed as a serum fetal defect/tumor marker to monitor disease progression. Besides this, AFP has many important physiological functions. As a tool of binding and transporting ligands, AFP may bind and transport fatty acids, steroids, heavy metal ions, drugs and some environmental contaminants. As a growth regulator, AFP is capable of both enhancement and inhibition. This function is dose-dependent. The regulatory mechanisms might involve apoptotic regulation, cytoplasmic signaling modulation, and receptor desensitization. As an anticancer drugs-ligand carrier, AFP can transport drugs into target tumor cells by binding AFP receptors overexpressed in the surface of tumor cells, and thus increase antitumor efficiency. In addition, AFP can be employed to chemoprevent breast cancer and possibly other tumors on the basis of its growth suppressive properties. PMID- 12561450 TI - Bad medicine. Why data from drug companies may be hard to swallow. PMID- 12561451 TI - Storm before the calm. Can knockout gases really be nonlethal? PMID- 12561452 TI - T cell triumph. Immunotherapy may have finally turned a corner. PMID- 12561453 TI - Scaled-up superposition. Supersizing Schrodinger's cat--by a billion times. PMID- 12561455 TI - Psychic drift. Why most scientists do not believe in ESP and psi phenomena. PMID- 12561454 TI - Reverse-engineering clinical biology. A peacetime dividend yields drug trials on virtual patients. PMID- 12561456 TI - Magnetars. PMID- 12561457 TI - New research addresses the wrenching question left when someone ends his or her own life. Why? The neuroscience of suicide. PMID- 12561458 TI - Evolving inventions. PMID- 12561460 TI - Satellite-guided munitions. PMID- 12561459 TI - Explaining frog deformities. PMID- 12561461 TI - Drink to your health? PMID- 12561462 TI - Synthetic diamonds. Carbon copy. PMID- 12561463 TI - Robots that suck. Have they finally come out with a robot for the rest of us? PMID- 12561464 TI - Why do some people get more cavities than others do? PMID- 12561465 TI - Why are snowflakes symmetrical? PMID- 12561466 TI - [Association of CYP2E1 of PON2311 polymorphisms in neonates with preterm]. AB - The objective is to investigate whether Rsa I polymorphism in the 5'-flanking region of CYP2E1 and PON2311 polymorphism in neonates are associated with preterm. Using standard questionnaires, 194 singleton live born mother-neonate pairs (including preterm cases and term controls) were investigated by the trained field workers with cross-sectional survey at the hospitals in Anqing, Anhui Province, China. Epidemiological and clinical data and blood samples were obtained from 194 mother-neonate pairs. CYP2E1 homozygous wild-type (cut/cut) is not associated with a shortened gestation among neonates, compared with CYP2E1 homozygous mutant-type (uncut/uncut)/CYP2E1 heterozygote (cut/uncut) before and after adjustment confounders. However, PON2 Ser311Ser homozygote is significantly associated with a shortened gestation among neonates. When Rsa I polymorphism in the 5'-flanking region of CYP2E1 and PON2311 polymorphism were considered jointly, a significant shortened gestation was observed among neonates with the combined genotype of CYP2E1 homozygous wild-type and PON2 Ser311Ser homozygote. In conclusion, Rsa I polymorphism in the 5'-flanking region of CYP2E1 in neonates is not associated with preterm, however, PON2311 polymorphism in neonates is significantly associated with preterm. Furthermore, the gene interaction between Rsa I polymorphism in the 5'-flanking region of CYP2E1 and PON2311 polymorphism in neonates is significantly associated with preterm. PMID- 12561467 TI - [Genetic polymorphism of six STR loci in the Han population in Henan province]. AB - To investigate the allele frequencies of six short tandem repeats (STR) loci: F13A1, F13B, D8S1179, CSF1PO, D5S818 and TPOX in Han population in Henan province, DNA was extracted with chelex from EDTA-blood samples of the unrelated individuals in Henan province and amplified with PCR technique. The PCR product was analyzed with the undenatured PAGE vertical electrophoresis and silverstain. The results were obtained through genetical analysis. The authors got the number of allele of six loci, Three alleles of F13A1 could be detected in this population. The F13A1 * 7 is the most common allele with a frequency of 45.2%, followed by F13A1 * 5 and F13A1 * 4 respectively. F13B had four alleles among which F13B * 10 has the highest frequency, followed by F13B * 9, F13B * 8, F13B * 11. The 8 alleles of D8S1179 in the order from high to low frequency is D8S1179 * 14, * 15, * 12, * 11, * 10. And the most common frequencies of alleles of CSF1PO, D5S818, TPOX ore CSF1PO * 12, D5S818 * 11, TPOX * 8, respectively followed by the allele of * 11, * 14, * 10, * 13 in CSF1PO locus, and * 12, * 13, * 10, * 9, * 14, * 8, * 15, * 7 in D5S818 locus, * 11, * 9, * 12, * 10 in TPOX locus. The heterozygosities of the six loci were 0.62, 0.46, 0.83, 0.59, 0.78 and 0.65; the discrimination powers were 0.78, 0.66, 0.95, 0.79, 0.92 and 0.82. The heterozygosities of the six loci are high and the frequencies are in good agreement with Hardy-Weinberg equilibrium. The six loci are good as genetic markers. We can use these loci in dividualidentification and partenity test in forensic area. PMID- 12561468 TI - [Establishment of mouse ZF-12+/- embryonic stem cells]. AB - By using the mouse zinc finger protein gene ZF-12 genomic DNA fragment, pSSC-TV 10.5 was designed and constructed as a replacement vector. Structure of pSSC-TV 10.5 was identified by restrictive digestion analysis and partly sequencing. Then linearized vector was electroporated into ES cells, and transfected cells were screened by G418 and GANC selection. Among 508 G418r/GANCr colonies, 4 were proved to have taken place the homologous recombination of ZF-12 by PCR and southern blotting analysis. This study lays the foundations of preparing mouse models of ZF-12+/- or ZF-12-/-. PMID- 12561469 TI - [The cloning and expression of a novel mouse gene mLPTS and its subcellular localization]. AB - A novel mouse gene mLPTS was cloned by EST assembling, RT-PCR and DNA sequencing. The gene fragment for mLPTS is 1244 bp in length, encoding a protein of 332 amino acids. The amino acid sequence of mLPTS has 78% homologue with that of LPTS gene, which is a novel liver cancer-related gene identified through positional candidate cloning stratage by our laboratory. The expression of LPTS gene was ubiquitous in normal human tissues, whereas levels appeared to be significantly reduced, or sometime undetectable in HCC cells and neoplastic tissues, and it might be involved in the negative regulation of cell proliferation. The expression of mLPTS gene was found in all mouse tissues analyzed, same with that of LPTS gene in human. There was only one transcript for mLPTS gene in mouse tissues. The phylogenetic tree was constructed through the amino acids sequence analysis and the study of the sequence homologue among different species. Next, mLPTS gene was cloned into green fluorescent protein eukarytic expression vector and then transfected into CHO cell line. The green fluorescent was mostly limited in the nucleolus, showing that the gene products of mLPTS in eukaryocytes were located in the nucleolus. PMID- 12561470 TI - [Isolation, localization and expression of a novel expressed sequence tag from pig skeletal muscular tissue]. AB - A novel expressed sequence tag (ESThp9-1, GenBank accession number: B1596262) was isolated from pig skeletal muscular tissue by using the mRNA differential display technique. BLAST analysis revealed that the 196 bp long EST (ESThp9-1) was not homologous to any of the known porcine genes in the database but similar to rat U3A small nuclear RNA (87% identity over 93 nucleotides) and mouse U3B.4 small nuclear RNA (85% identity over 96 nucleotides). Semi-quantitative reverse transcription polymerase chain reaction indicated that EST9hp-1 was expressed in most of tissue of the pig. ESThp9-1 was physically mapped on sus scrofa chromosome 12q1.1-q1.5 and linked with microsatellite S0090 by using somatic cell hybrid panel and radiation hybrid panel analysis. According to the homologous information and result of physical mapping, ESThp9-1 was presumed to be one member of the porcine U3 gene family. PMID- 12561471 TI - [Analysis and mapping of homologous sequences of barley disease resistance gene Mlo and maize disease resistance gene Hm1 in rice]. AB - The encoding products of the disease resistance (R) gene of barley, Mlo, and the maize R gene, Hm1, do not contain the recognizable structural domains of most of the known plant R gene products. The two genes do not comply with the gene-for gene theory in the response to pathogen infection either. Two sequences, OsMlo-1 and DFR-1 which were homologous to the barley Mlo and maize Hm1, respectively, were isolated from rice. The OsMlo-1 was mapped between two molecular markers, RZ667 and RG424, and located at a distance of 20.6 centi-Morgan (cM) from RZ667 and 6.0 cM from RG424 on rice chromosome 6. The DFR-1 was flanked by molecular markers R2635 and RG462 with an estimated genetic distance of 11.3 cM from R2635 and 23.9 cM from RG462 on rice chromosome 1. Using other published rice molecular linkage maps as references, it was found that the chromosomal locations of OsMlo 1 and DFR-1 were correspondence with two previously identified rice blast resistant QTLs (quantitative trait loci), respectively. The results suggest that the genes homologous to barley Mlo and maize Hm1 in rice may be involved in defense responses induced by pathogen infection. PMID- 12561472 TI - [Genetic mapping of T-DNA integration sites in Xa21 transgenic rice]. AB - The transformation mediated by Agrobacterium has been successfully applied to rice in recent years. In the previous research we have transferred the Xa21 gene into five rice varieties of China, using Agrobacterium-mediated trasformation. In this study, T-DNA flanking sequences of Xa21 transgenic rice lines were obtained by using thermal asymmetric interlaced PCR (TAIL-PCR). The flanking sequences which are actual rice DNA were identified and located on molecular linkage map developed from a ZYQ8/JX17 double haploid (DH) population. A total of 22 T-DNA flanking rice sequences were isolated. Nineteen of them displayed RFLPs between the two parents, ZYQ8 and JX17, and were mapped on the rice chromosomes, 3, 4, 7, 9, 10, 11 and 12, respectively. The genetic mapping of T-DNA integration sites in Xa21 transgenic rice will benefit the study of position effect and stable inheritance of the transgene Xa21. PMID- 12561473 TI - [Analysis of gene expression profiles during host-Magnaporthe grisea interactions in a pair of near isogenic lines of rice]. AB - A pair of near isogenic lines G205 and G71 were selected from recombinant inbred lines (RIL) of Zhong156 x Gumei2. On the resistance locus Pi-25(t), G205 had the resistant allele that was from Gumei 2 while G71 had the susceptible allele that was from Zhong156. For the genetic background, different alleles were detected on only 24 loci out of the 672 RFLP or SSLP loci surveyed. The expression profiles of G205 and G71 in response to Magnaporthe grisea were investigated using cDNA microarray containing 2200 Expression Sequence Tags (ESTs). The leaves were inoculated with the pathogen for 12 hours at 4-leaf stage and 998 genes were identified in total. Three genes were up-regulated significantly by the fungus in G205 only. The functions of two genes were known but that of the third gene were unknown. The two genes encoded casein kinase II alpha subunit and retrotransponson TOS17 insertion element respectively. Other thirty-five genes had similar expression patterns between NILs. Among them, 17 genes were up regulated while 18 genes were down-regulated by the inoculation. The functions of 33 out of the 35 genes were known. BLAST analysis showed that all thirty-five. BLAST analysis showed that all thirty-five genes with known functions were relative to defense reactions, signal transduction, stress response, photosynthesis and sugar metabolism. Northern blot confirmed that four of five differentially displayed genes randomly selected had the same expression patterns as those detected in cDNA microarray. Two of them were up-regulated genes encoding casein kinase II alpha subunit and glycine-rich protein (Grp), and the other two down-regulated genes encoding nitrilase-associated protein and 18S small subnit ribosomal RNA gene respectively. Northern blot also revealed that the expression of Grp was consistently up-regulated from 0 to 36 h after the inoculation of the fungus. These results showed that cDNA microarray was a useful tool to study the molecular mechanisms of disease resistance in plants. PMID- 12561474 TI - [Construction and analysis of near-isogenic line derived from wheat cultivar sumai 3]. AB - Wheat scab can cause significant yield lost and quality decrease as well as toxicoses in animals and humans. Sumai 3, a resistant cultivatr to scab, is widely used in the wheat breeding of scab resistance. To study the inheritance of resistance to scab in Sumai 3, the highly susceptible cultivar Chuan 980 was crossed with Sumai 3 and backcrossed with Sumai 3 as a recurrent parent for seven times, thus a near-isogenic line S016 susceptible to scab was developed. S016 was evaluated in five regions and a period of two years for resistance to spread of scab in spike. Results showed that S016 was as highly susceptible to wheat scab as one of the parent Chuan 980. Molecular markers (RFLP, RAPD) were screened to identify chromosome regions of negative element of resistance gene (s) in S016. The DNA polymorphism between the near-isogenic lines was showed using six restrict enzymes (EcoRI, EcoRV, HindIII, Dra I, BamH I, Xba I) and 85 probes located in wheat chromosome 2D. Among the amplified bands of 450 10-mer random primers, OPH191400 and OPH191200 were perhaps linked to negative regulator element. Some reported probes linking to scab resistance gene(s) did not show any polymorphism between this near-isogenic line and their generation. PMID- 12561475 TI - [Studies of the relationship between the breeding efficiency of anther culture and the use of different hybrid generations as anther-donor plants in wheat]. AB - The wheat haploid plantlets from anther culture were of theoretical and applied interest. Induced haploid spirophytes may play a key role, not only in increasing the efficiency of breeding programs by producing pure lines rapidly, but also in the elucidation of fundamental genetics. However large number of experiments turned out that there are many problems with low rate of green plantlet from anther culture, difficulties of saving plantlets though summer, low rate of double chromosome, extrusive contradiction between technique of wheat anther culture and variety improvement. So wheat anther culture has not played a role in wheat breeding. Studies were carried out on the relationship between the breeding efficiency of anther culture and the use of different hybrid generations as anther-donor plants in wheat. It was shown that previously the low breeding efficiency of anther culture in wheat cross breeding was mainly due to the use of F1 as anther donors. Experimental results demonstrated that the breeding efficiency of anther culture in F1 generation was very low, especially when the doubled haploid population was very small. Based on the experimental results as well as the theoretical analysis this paper proposes that the use of F2 or F3 (especially F3) as anther donors in combination with the conventional selection in the fields will increase the breeding efficiency of anther culture. Under the guidance of this proposal two wheat varieties have been developed within 5-7 years through anther culture. PMID- 12561476 TI - [Inheritance of resistance of southern corn rust in the inbred line Qi319]. AB - Southern corn rust (SCR) is a destructive disease in maize. The inbred line Qi319 is highly resistant to southern corn rust. The inheritance of resistance to southern rust in Qi319 was investigated. Five F1 hybrids were derived from Qi319 crossed with five susceptible inbred lines respectively. The F2 generations were produced by F1 self-pollinated and BC1 F1 generations were abtained by backcrossing F1 with the susceptible parents. Inoculation of the P1, P2, F1 s and 10 individuals of F2, BC1 F1 were completed with the southern corn rust pathogen and showed that all of the 10 F1 s were resistant, the all 5 F2 populations segregated in a ratio of 3R:1S, and all of the 5 BC1 F1 populations segregated in a ratio of 1R:1S. Therefore, it is considered that Qi319 carries one dominant gene for resistance to southern corn rust. PMID- 12561477 TI - [Sequence variation of the chloroplast gene ndh D region in cytoplasmic male sterile sorghum]. AB - A fragment SAAU-02(700) was amplified specifically from total DNA of seven sorghun varieties with male-fertile cytoplasm (N-cytoplasm). PCR assays indicated that it was amplified from chloroplast (cp) DNA. Sequence analysis revealed this newly cloned fragment contained a portion of chloroplast gene psa C (88 bp) and part of ndh D gene (192 bp). Total DNA, mitochondrial (mt) DNA, and cpDNA were digested with EcoR I + Hind III and probed with fragment SAAU-02(700). The Southern hybridization patterns displayed a 0.74 kb band both in total DNA and cpDNA, but an additional faint band 0.45 kb in size was found only in the latter. No polymorphic hybridization signal between the N-cytoplasm and male-sterile cytoplasm (S-cytoplasm) was observed. Southern hybridization of total DNA of CMS line A1 Tx623 and fertile line Tx623 digested with Hae III gave a band 4.9 kb in size in the former and a 4.45 kb band in the latter. This revealed that the sequence of ndh D from CMS line was likely altered. Further studies designed to determine whether or not the variation has some effect on the metabolism of mitochondria and chloroplast, even on the occurrence of male sterility in sorghum are underway. PMID- 12561478 TI - [Analysis of the inter-species relationships on lycoris Amaryllidaceae) by use of RAPD]. AB - The fingerprints of 13 species in genus Lycoris were generated by use of RAPD method. Forty-one primers were screened from 520 random primers, and a total of 350 DNA fragments were amplified ranging from 0.3-3.0 kb, 253 (72.3%) of which were polymorphic. The average number of DNA band produced by each primer was 6.2. Nei's similarity coefficients and genetic distances were calculated by use of the software of TFPGA version 1.3 and dendrogram of Lycoris was constructed using UPGMA. It is indicated that the 13 species of the genus Lycoris were divided into two groups, and five species of the genus including L. rosea, L. haywardii, L. straminea, L. sprengeri and L. radiata with monotype karyotypes (I-shaped) were clustered together respectively. The basic chromosome number was x = 11. The others which have two-types karyotypes (I-shaped and V-shaped) were clustered together respectively. They were L. houdyshelii, L. albiflora, L. chinensis, L. longituba, L. anhuiensis, L. squmigera, L. caldwellii and L. aurea. The closest relationship was between L. rosea and L. haywardii. L. radiata is highly divergent from L. aurea. The results were in consistence with that of the analysis of chromosome karyotype. The present paper discussed the problems whether L. rosea, L. haywardii and L. stramina originated as natural hybrids and taxonomy position of L. albiflora, L. straminea and L. houdyshelii based on the RAPD analysis. PMID- 12561479 TI - [Specific molecular markers of the rust resistance gene M4 in flax]. AB - Flax (Linum usitatissimum L.) is an important fiber and oil-producing crop. Flax rust, caused by Melampsora lini Ehrenb. Lev., occurs worldwide and can cause severe losses in seed yield and fiber quality. In order to identify molecular markers linked to the flax rust resistant gene M4, RAPD analysis of NM4, a near isogenic line containing the M4 gene, and the recurrent parent Bison was carried out with 540 decamer primers. The primer OPA18 could stably amplify a specific fragment, OPA18(432), in the NM4 line. The OPA18(432) marker was testified to be closely linked to the M4 gene with a genetic distance of 2.1 cM through the analysis of the F2 mapping population derived from a cross of Bison x NM4. Based on the sequence of OPA18(432), the specific PCR primers were designed, and a SCAR marker for the M4 gene was produced. Amplification of different resistant materials proved that the maker is specific for the M4 gene. This marker has been used successfully in marker-assisted selection in the flax breeding program. PMID- 12561480 TI - [Quantitative genetic analysis and multiple trait selection of pharmaceutical composition on Ginkgo biloba L. leaves]. AB - The China possesses more than 70% Ginkgo resources in the world. The purposes of this research are to collect fine seedling and grafting clones in China, to probe genetic law on flavone glycosides and terpenes on Ginkgo leaves, and select leaf used cultivars of high-pharmaceutical composition. In this research, we have collected 87 clones from 13 provenances and have carried out randomized block experiment at Tancheng, Laizhou and Yaoxiang in Shandong Province. Flavone glycosides and terpenes were determined through HPLC method from an improved Van Beek (1991) techniques and Shimadzu Lc-10 AD (Japan). Data and breeding analyses were carried out through IBMPC and SPQG30 software. The results of variance analyses show that there are significant differences to flavone glycosides, terpenes in clones, and the law of genetic parameters on heritability (h2) and genetic variability coefficient (Gcv), is clone > sex > provenance to flavone glycosides in ginkgo leaves. The sigma g2, h2, Gcv and delta G' in male tree clone leaves are higher than female clone leaves on flavone glycosides. We have found that there is a maximum flavone content clone among males and a maximum terpene clone among females. The results of Q-cluster analyses are consistent with R-factor analyses of twenty higher terpenes clones. The results of index selection show that the ri.Y2, E(I) and CGS' of multiple traits selection including (gamma) trait are higher than single trait and multiple traits selection excluding gamma. The direct or index selection is more suited to leaf used cultivars of Ginkgo. The genetic stability of each clone was appraised by Wricke's ecovalence method and Nassar & Huhu noparameter method. Flavone glycosides and terpenes are more than 2.09%-2.57% and 0.33%-0.41%, respectively, and we have selected four clones. PMID- 12561481 TI - [Analysis of gene cluster of Tat-dependent protein export system of Vibrio cholerae and its function]. AB - The Tat (Twin-arginine translocatin) system is a recently defined protein export pathway that serves to translocate folded proteins. The substrates of the Tat pathway contain specific amino-terminal signal peptides that exhibit a conserved amino acid consensus motif-S/T-R-R-x-F-L-X. Here is the report of knocked out the tatA, tatB and tatC genes of the V. cholerae by suicide plasmid homologous recombination technology. Mutant strains showed obvious changes of growth characteristics. The transport of a typical Tat pathway substrate, trimethylamine N-oxide reductase (molybdoenzyme), was completely blocked. The physiochemical reactions of the parent and mutant strains were also analyzed. Four physiochemical reactions using D-galactose, L-asparagine, glyl-L-aspartic acid and D-L-alpha-glycerol phosphate as substrates were negative in mutant strains, which might be affected by the inactivation of the Tat-dependent system. PMID- 12561482 TI - Putting the emphasis on employees. AB - Baptist Health Care Corp., Pensacola, Fla., has been racking up awards and kudos for its employee-friendly culture. This environment has boosted patient satisfaction and staff retention dramatically while also improving quality of care. PMID- 12561483 TI - 'Concierge' medicine. Who really pays for gold standard access to doctors? AB - Unhappy with health plans that limit their patient contact time and require too much paperwork, some doctors are joining groups that require patients to pay a membership fee for "gold-standard" care. Experts weigh in on this trend, its future and its ethics. PMID- 12561484 TI - Trustee workbook 1. Trends in board accountability: conducting the audit. PMID- 12561485 TI - Forward motion. Interview by Mary Grayson. AB - AHA chair-elect, David L. Bernd, CEO of Sentara Healthcare, Norfolk, Va., discusses the current state of health care and his hopes for the future. PMID- 12561486 TI - So, greed's not good after all. PMID- 12561487 TI - Maximizing construction value through a design competition. PMID- 12561488 TI - Opening doors to patient safety: a board checklist. PMID- 12561490 TI - Brothers in arms. PMID- 12561489 TI - State of emergency. PMID- 12561491 TI - Primary care. More than words. AB - There are few models for involving patients in primary care. Research in one primary care group, involving 92 patients from nine general practices, established a list of 20 standards most important to patients. When asked to assess themselves against these standards, practices rated their services higher than patients did. The research found considerable resistance to patient involvement from GPs, which needs to be addressed if the government's aims are to be achieved. PMID- 12561492 TI - Primary care. Central reservations. AB - A primary care trust has established a call centre for booking appointments for community services. The service handles bookings for physiotherapy, minor surgery and retinal screening. Since it went live in April 2002, the service has booked 2,000 appointments, reduced waiting times for physiotherapy appointments from 16 to four weeks and cut non-attendance rates from 18 to 2 per cent. It is hoped to increase operating hours to 7.30 pm and extend to cover other services. PMID- 12561493 TI - First person. Open and shut case. PMID- 12561494 TI - UCLA expands automatic patient data collection system. PMID- 12561496 TI - Electronic signature system speeds up data preparation. PMID- 12561495 TI - Michigan hospitals polish data to qualify for payment bonuses. PMID- 12561497 TI - Data flow easily between departments at Florida Hospital in Orlando. PMID- 12561498 TI - Hospitals predict market growth with the help of public databases. PMID- 12561499 TI - Annual risk of tuberculous infection in rural areas of Junagadh district. AB - Paucity of epidemiological data on tuberculosis in India prompted the National Tuberculosis Institute, Bangalore to embark upon a nation-wide survey to estimate the Annual Risk of Tuberculosis Infection in different parts of the country. The survey in Junagadh district, one of the 26 districts drafted under the nation wide survey is reported here. The prevalence of infection from the analysis of 3164 children not displaying scars of the BCG vaccination was 4.16% (CI: 3.17 5.14) and from this data the ARI was computed as 0.73% (CI: 0.55-0.91). The inclusion of vaccinated children into the study group yielded similar results. The estimate of the ARI in Junagadh district is lesser than that in several other parts of India which is probably in consonance with its better socio-economic development. PMID- 12561500 TI - Detection of tubercle bacilli in sputum: application of sodium hypochlorite concentration method. AB - Tuberculosis has emerged as the leading cause of death due to single infectious agent. Sputum microscopy forms mainstay of diagnosis as it is most reliable, specific and objective method available especially in developing countries. However, this method is hampered by lack of sensitivity. Liquefaction and concentration of sputum before Ziehl Neelsen staining improves yield and also makes examination of smears rapid and convenient. A total of 304 sputum samples were studied by direct staining and after sodium hypochlorite treatment and centrifugation. Direct smears stained with Auramine-Phenol were also examined. Use of sodium hypochlorite method increased the number of positive samples from 52 to 96. No false positives were detected by either method. The method is simple as the only extra reagent required is sodium hypochlorite that is easily available. Additional advantage is that sodium hypochlorite, being a potent disinfectant, reduces the risk of laboratory acquired infections. PMID- 12561501 TI - A community survey of dog bites, anti-rabies treatment, rabies and dog population management in Bangalore city. AB - This survey was conducted by using a 30 cluster random sampling technique to comprehensively study the nature and magnitude of rabies and its related problems in human and dog population. The city has an estimated dog population of 3.25 lakhs of which 2 lakhs (61.5%) are stray dogs and 1.25 lakhs (38.5%) are pet dogs. The dog:man ratio was about 1:12 and the pet:stray dog ratio of 1:2. The awareness about Animal Birth Control (ABC) programme was low (34%) and only 20% had faith in it. The annual incidence of dog bites was 1.9% persons. It was more in males (64%) and mostly from stray dogs (64%). About 86% of them received anti rabies vaccination and none received life saving Anti-Rabies Serum (ARS). The annual incidence of human rabies was 15 and dog rabies 58 resulting in a ratio of 1:4 approximately as a rabies contagion index. The implementation of ABC programme was weak (10.4% coverage) and is recommended for acceleration and improvement. Similarly modern anti-rabies immunization, including anti-rabic serum, shall replace the outdated sample (sheep brain) vaccine. PMID- 12561502 TI - Rapid diagnosis of Mycobacterium tuberculosis by multiplex polymerase chain reaction from clinical specimens. AB - An essential element in the control of tuberculosis is the rapid, sensitive and specific identification of the causative agent. Until now, screening and diagnosis are largely based on clinical signs, radiological examination, tuberculin tests, sputum examination under the microscope, or culture for mycobacteria. Tuberculin tests lack specificity and only give an indication of previous exposure to mycobacteria. Direct microscopic examination of sputum is neither specific nor sensitive enough, and mycobacterial isolation is time consuming. As an alternative to these classical methods, new nucleic acid-based technologies show promise as a more rapid, sensitive, and specific means of identification of mycobacteria. Two commercial standardized nucleic acid-based amplification techniques have been reported to yield reliable results within 5 to 7 hrs. Roche Amplicor MTB (Roche Diagnostic System, Somerville, N.J.) and Gen Probe AMTB (Gen-Probe Inc., San Diego, Calif.). The amplified target is part of the 16S rRNA gene which is common to all the mycobacteria. An attempt has been made to describe the use of the target DNA, SenX3-RegX3, in a multiplex PCR to detect and differentiate M. tuberculosis from other mycobacteria directly from clinical specimens. PMID- 12561503 TI - A study on the status of Japanese encephalitis infection in Arunachal Pradesh. AB - Suspected cases of Japanese Encephalitis (J.E.) were recorded in the hospitals of Arunachal Pradesh from 1986 to 1995. 162 cases were diagnosed as J.E. with predominance in male sex and lower age group. Maximum cases were recorded between June to October. 11.3% pig sera showed J.E. antibodies when tested by ELISA test. PMID- 12561504 TI - Animal bite management practices: a survey of health care providers in a community development block of Haryana. AB - It is seen that outcome of animal bites is influenced by various factors including the treatment procedures practiced by health care providers (HCPs). A cross sectional study of health care providers was conducted during May 2000 in PHC Kurali and Naraingarh town of community development block Naraingarh in district Ambala, Haryana. A total of forty-four HCPs were interviewed at their health facility. They were asked about the qualification and number of years in practice. Health care providers were assessed for their knowledge regarding history taking, immediate management of animal bite, post bite anti-rabies treatment, follow up advice and availability of vaccines. Pre-exposure prophylaxis was known to 18.8% of HCPs. Fifty-nine per cent of HCPs were confident in managing dog bites and 93.1% knew about tissue culture vaccine. Vaccine cost was the commonest barrier (38.8%) in the management of animal bites. This study shows a gross difference between awareness and actual practice of management of animal bites. PMID- 12561505 TI - An outbreak of dengue fever in rural areas of northern India. AB - During the past few decades, epidemics of dengue fever are causing concern in several South-East Asian countries including India. The rural areas of Hissar district of Haryana state, situated about 170 km North-West of Delhi, experienced an outbreak of febrile illness during July-August 1996. A total of 13 villages in eight affected primary health centres reported fever cases. The clinical, epidemiological and entomological findings indicated that the present episode was due to dengue fever. The aetiological agent of the current outbreak, the DEN-2 virus, was isolated from 12 acute-phase sera specimens. Though, in the recent past outbreaks have been reported from the rural areas of southern and western India, the present episode is the first outbreak being reported from the rural areas of northern India. The increasing frequency of dengue fever outbreaks in rural areas of various Indian states reflects the changing life style of the rural population as a result of urbanization process and calls for a suitable prevention and control policy based on strengthened surveillance, appropriate health education to the community coupled with proper training of health personnel. PMID- 12561506 TI - Epidemiology of cholera--a five year study. AB - A total of 286 strains of Vibro Cholerae were isolated and tested over a period of five years. The strains were identified by standard methods and confirmed by slide agglutination tests with polyvalent, Ogawa and Inaba antisera. The non agglutinating strains were tested with O-139 antisera. The maximum number of cases were found in the age group of 0-10 years. The number of females affected was more than the males. V. cholerae O-139 was isolated in the year 1998 and then again in 2000. V. cholerae serotype Inaba was found only in the year 1999. All of the other isolates belonged to the serotype Ogawa. The periodic shift between O1 and O-139 serogoups is reminiscent of the shifts from the Ogawa to the Inaba serotypes periodically witnessed among V. cholerae, possibly mediated by the immune pressure in the population. PMID- 12561507 TI - Knowledge amongst adult population regarding vectors of malaria in 21 states of India. AB - Malaria is the world's most important tropical disease which kills more people than any other disease except tuberculosis. It is a public health problem in more than 90 countries, inhabited by a total of some 2400 million people, 40% of the world's population. More than 90% of all malaria cases are in sub Saharan Africa, with two thirds of the remainder concentrated in six countries viz. India, Brazil, Sri Lanka, Afghanistan, Vietnam and Columbia in decreasing order of prevalence. Even now the problem of malaria in India is grossly underestimated. A rough estimate of morbidity due to malaria made on the basis of consumption of antimalarials comes to 35.5 million episodes in addition to malaria cases treated by the National Anti Malaria Programme (NAMP). In addition to large scale morbidity and mortality, it affects agricultural and industrial produce causing great socioeconomic losses. PMID- 12561508 TI - Prevalence of intestinal parasitic infections among patients attending Adichunchanagiri Hospital and Research Centre, B.G. Nagar, Mandya (Karnataka). PMID- 12561509 TI - [Studies on the specificity of target organs and repair dynamics of DNA damage induced by 1,2-dichloroethane]. AB - In order to investigate the genotoxicity of 1,2-dichloroethane (DCE) and to determine its carcinogenic targets, the DNA damage and repair dynamics were detected by using comet assay in the isolated nuclei with tissue-homogenization. Liver, lung, kidney, spleen, bone marrow, testicle, stomach, colon, bladder, and peripheral lymphocytes of mice were isolated at 3 h, 8 h, 24 h after DCE being gavaged. The results showed that: a) liver, lung, kidney, bone marrow, colon, stomach and peripheral lymphocytes were the genotoxic target organs of DCE in mice; b) DNA damage and repair dynamics were obviously different among the test organs, and the organs with DNA damage and slow repaired (lung, stomach and hemocyte) were in accordance with the carcinogenic targets. The results indicate that multiple organ comet assay in vivo is very useful for the evaluation of genotoxicity and prediction of carcinogenic target organ. The comet tail of peripheral lymphocyte is suitable as a biomarker for DNA damage in the target organs of living creatures and population exposed to DCE. It is suggested that in vivo comet assay is also very important to select right marker cells and biomarker in the epidemiological studies on genotoxicity. PMID- 12561510 TI - [Effect of cadmium on the phagocytic function of polymorphonuclear leukocyte in mice tested by chemiluminescence of whole blood]. AB - The effect of cadmium chloride on the immunological toxicity of polymorphonuclear leukocyte (PMN) and on lipid peroxidation in liver were measured by using the chemiluminesence of whole blood and thiobarbituric acid (TBA) method. The results showed that 0.1 and 10 mmol/L of cadmium chloride could accelerate the chemiluminesence of whole blood in vitro, but 1000 mmol/L of cadmium chloride inhibit the chemiluminesence. Two hours after the injection of cadmium chloride through caudal vein, chemiluminesence of whole blood was induced obviously at the dose of cadmium 1 and 100 micrograms/kg BW in vivo, but the chemiluminesence was inhibited at the dose of 500 and 1000 micrograms/kg BW. At the dose of 1 microgram/kg BW, the peroxidation of liver lipids was induced. The results suggested that cadmium at low concentration accelerated the immunity of PMN, while high concentration was toxic to PMN, cadmium at low concentration could accelerate the phagocytic function of phagocytes. Phagocytosis brings about excessive active oxygen in organism and intensifies the action of active oxygen on the lipid peroxidation in tissue cells, at last, results in organic oxidation injury. PMID- 12561511 TI - [Study on the form and stability of alpha and beta domain of metallothionein binding to lead (II)]. AB - For studying the actions of metallothionein(MT) binding to lead, the alpha and beta structural domain of metallothionein binding to Cd (Cd7-alpha-MT and Cd7 beta-MT) was extracted and purified from a strain of BL2I by dissolving them in acid to eliminate protein. The apo-alpha-MT and apo-beta-MT were separated by gel Sephadex-G25 to eliminate Cd. Different doses of lead were added to the apo-alpha MT and apo-beta-MT. The structure of MT metal Cluster with lead was formed after reacting completely, and was then examined by ultraviolet spectrophotometer. The UV spectrum showed that the interaction of lead with alpha domain leading to two kind of products, one binding with 4 lead(Pb4-alpha-MT) and the other binding with 7 lead (Pb7-alpha-MT). Only one product, the beta domain binding with 3 lead(Pb3-beta-MT) was formed. The reaction rate of these products with DTNB was also measured in this study. Results showed that the domain binding to 4 lead was more stable than the others. All these works provide strong evidence that further studies on the mechanism of MT interacting with lead and the function of MT in eliminating lead were needed. PMID- 12561512 TI - [Effects of microwave radiation on the content of five elements in mice bone tissue]. AB - Mice were radiated with 2450 MHz, 10 mW/cm2 microwave for 12 days, 1.5 h/day. After microwave radiation, compared with the normal control, the content of calcium and zinc in mice bone were significantly decreased (P < 0.05) copper, iron and manganese decreased, appulsively After Libido, a composed traditional herb medicine, and asshide asafetida were supplied seperatively, the content of calcium and trace element zinc in mice bone increased (P < 0.05). It is concluded that Libido was effective on the resistance of mice to microwave radiation. The toxicity of organotin compounds and the current pollution status. PMID- 12561513 TI - [Effects of cactus, alove veral, momorcica charantia on reducing the blood glucose of diabetic mice]. AB - The effects of cactus, alove veral and momorcica charantia on reducing the blood glucose level of mice were observed. The diabetic model with no symptom in mice was established by injection of streptozotocin(STZ) 80 mg/kg BW into abdominal cavity for 11 days. The diabetic mice were randomly divided into 8 groups: STZ diabetic model, diet A, diet B, cactus, alove veral, momordica charantia and glyburide groups. Cactus (60 g/kg BW), alove veral (60 g/kg BW), and momordica charantia (30 g/kg BW) were administrated orally each day to the diabetic mice for another 21 days. Serum glucose of mice fasting for 12 hours and 2 hours after meal was determined with the method of glucose-oxidase at the 21th day of the experiment. The results showed that serum glucose levels of diabetic mice were significantly higher than the normal control group (P < 0.01). After giving diet A, cactus, alove veral and momorcica charantia juice for 21 days, the serum glucose concentration of these diabetic mice were significantly lower than STZ diabetic model group (P < 0.01) but still higher than the normal control group. PMID- 12561514 TI - [Study on the hypotensive effect of manascus]. AB - The hypotensive effect of manascus was studied by using three well established rat models with hypertension: spontaneously hypertensive rat (SHR), renalvascular hypertensive rat (RHR, 2K1C) and DOCA-salt hypertensive rat. Manascus of 0.0, 0.4, 0.8, 1.2 g/kg BW (p.o.) were administrated daily to these animals for 3-4 weeks. Systolic blood pressure (SBP) and heart rate were measured once a week by using tail-cuff method. The hypotensive effect of manascus was observed in SHR and DOCA-salt hypertensive rats; and the effect was more potent in DOCA-salt hypertensive rat. Manascus showed less potential hypotensive effect in RHR. PMID- 12561515 TI - [Studies on the hydroxyl free radical-scavenging effect of combined selenium and germanium]. AB - The effect of selenium, carboxyethyl-germanium sesquioxide (Ge-132) and the combination of selenium and Ge-132 on the production of hydroxyl free radical in liver microsomes of rats treated with Fe2SO4-NADPH system was studied with electron spin resonance technique (ESR). The results showed that the production of hydroxyl free radical was decreased significantly by adding selenium, Ge-132 and combined selenium and Ge-132, indicating a direct scavenging effect on hydroxyl free radical. It was also observed a enhanced scavenging effect at the low concentration of combined selenium and Ge-132. PMID- 12561516 TI - [Effects of olive oil and fish oil on serum lipids and lipid peroxidation in rats]. AB - The effects of olive oil and fish oil on serum lipids and lipid peroxidation were studied in rats. The rats were divided into four groups: negative control, high fat control, fish oil (high fat diet and tube feeding with fish oil 4 ml.kg-1.d 1) and olive oil (high fat diet and tube feeding with olive oil 4 ml.kg-1.d-1) groups. The results showed that the total serum cholesterol and the triglyceride decreased significantly both in fish oil group and olive oil group, and there was a significant increase of serum high-density-lipoprotein (HDL) cholesterol in fish oil group. Both fish oil and olive oil increased superoxide dismutase (SOD) in erythrocyte and decreased serum MDA. PMID- 12561517 TI - [Inhibitory effects of beta-carotene on hepatic cancer cell line SMMC-7721]. AB - The inhibitory effect of beta-carotene on the proliferation of hepatic cancer was studied. Cells from a hepatic cancer cell line SMMC-7721 were incubated in culture media with 20, 40 and 80 mumol/L beta-carotene for 12, 24 and 48 h respectively. MTT test, Trypan blue exclusion test and DNA gel electrophoresis were used. The results of MTT test revealed that beta-carotene (20-80 mumol/L) could inhibit the proliferation of SMMC-7721 cells in a dose-dependent manner. DNA gel electrophoresis showed that the apoptosis of hepatic cancer cells could be induced by beta-carotene (40 mumol/L). It is concluded that the proliferation of hepatic cancer cells inhibited by beta-carotene, probably through interfering DNA metabolism and inducing cell apoptosis. PMID- 12561518 TI - [Antioxidation of flavones of wheat germ on mammary tumor of rats]. AB - The effect of flavonoids of wheat germ on mammary tumor of rats induced by 7,12 dimethylben(a) anthracene (DMBA) was investigated. Sprague-Dawley female rats (50 day-old, weighted around 176 g) were randomly divided into 4 groups. The negative and positive control group were fed on stoke diet. The high and low dose test groups were fed on diets with wheat germ flavonoids 10 and 2 g/kg respectively. Except rats in the negative control group, each rat was given DMBA 15 mg dissolved in 1.5 ml vegetable oil by tube feeding. After the administration of DMBA for 24 weeks, the incidence of tumor in the high dose test group was lower than that in the positive control group. The activity of blood and liver glutathione peroxidase (GSH-Px) and peroxidase dismutase(SOD) in the test groups was significantly higher than those in the positive control group, while the MDA level was significantly lower. The results suggested that the effect of flavonoids on inducing peroxidase might be one of the chemical prevention mechanisms on mammary tumors. PMID- 12561519 TI - [Establishment of fetal limb bud culture model of mouse]. AB - In order to study the development of bone and the effect of each single factor on the metabolism of bone during gestation age, a self-made rotating apparatus was designed. The limb bud of fetal mouse at the 16th day of gestation was examined with the techniques of X-roentgenography and histological slices. Comparing with the uncultued control limbs, the test limb cultured for six days showed longer bone length and higher bone density. The cultured bone cells showed active differentiation, proliferation and increased bone trabecula by histological examination. It is concluded that the cultured bone continued developing and differentiating, which confirming the feasibility and reliability of the method. PMID- 12561521 TI - [Transition of dietary pattern in China]. AB - The implications of dietary transition in China during the past 50 years were investigated. Data were collected from the representative national surveys, China Health and Nutrition Survey (1989-1997), China National Nutrition Survey (1982 and 1992), the Annual Household Consumption Surveys of the State Statistic Bureau and the Annual Death Report of China. In the early part of the major economic transformation, cereal intake increased before 1985 and decreased thereafter. There was also a long-term reduction of vegetable consumption, which has stabilized now. Intakes of animal foods increased slowly before 1979 and more quickly after the economic reforms occurred. The total energy intake has reduced, as energy expenditure has, large changes in the composition of energy have occurred. The proportion of energy from fat increased quickly and reached 27.3% in average and 32.8% in urban residents in 1997. More than one third of all Chinese adults and 58.4% of adults in urban areas consuming over 30% of their energy from fat. These changes resulted in rapid increases the prevalence of overweight and obesity and dietary-related chronic non-communicable diseases in urban residents. The traditional Chinese diet is shifting towards a diet with high fat, high energy density and low dietary fiber. The Chinese diets have entered into a new stage of transition. PMID- 12561520 TI - [Changes and relations of leptin, growth hormone and insulin during puberty in obese and non-obese children]. AB - In order to study the regularity of changing leptin, growth hormone (GH) and insulin (INS) during puberty and their relationships, a group of 300 obese and 300 non-obese children, aged 10-15 were selected. Leptin level increased with age and then decreased in boys, but it had only increasing tendency in girls. The GH level increased and then decreased suddenly in all groups. There was no obvious regularity in the change of INS. Serum leptin and INS levels were higher (P < 0.01) in obese children than that in non-obese children, while GH levels were significantly lower (P < 0.01) in obese boys. Results also showed that GH was negatively correlated with leptin in boys (obese group: r = -0.74, P < 0.05; non obese group: r = -0.69, P < 0.05) and positively correlated with leptin in girls (obese group: r = 0.58, P < 0.05; non-obese group: r = 0.67, P < 0.01). There was a positive correlation between INS and leptin in non-obese girls (r = 0.54, P < 0.05). It is concluded that leptin might play an important role during the initial stage of puberty in children and the effect of leptin on pubescent development in girls is greater compared with boys. The correlation between leptin and GH are gender dependent in boys and girls, which may cause the timing differences of sudden growth in boys and girls. PMID- 12561522 TI - [An analysis on the forearm bone mass density of rural female and the environmental risk factors]. AB - The distribution of distal and proximal forearm bone mass densities (BMD) with age was discribed and the environmental risk factors of rural female analyzed. A group of 1432 rural female aged 15 and over were sellected. Their demographic characteristics, living and eating habit were obtained by standardized questionnaire. The distal and proximal forearm bone mass density were measured by peripheral dual-energy X ray absorptionmetry (pDEXA). The results showed that the distal and proximal forearm BMDs were increased with age before age 25 and 30 respectively, and reached the peak value at age 30-35. The distal forearm bone density decreased significantly at age 40 while the proximal forearm BMD decreased at age 45. Bone loss rate of the two bone sites was increased significantly at age 50 and reached the peak value at age 55-60. Only the the density of proximal forearm bone, and the year of menopause was the main cause of low bone density. Body weight was the positive factor for bone density at age less than 60. Height only positively affected the proximal forearm bone of those at age 30-45. More ever, drinking tea, parity and educational status may affect distal forearm bone in certain age group while parity, educational status, occupation and marital status were possible risk factors of proximal forearm BMD. It is concluded that environmental risk factors of BMD varied with bone site and age. The prevention of low BMD must rely on subject's age and bone site. The surveillance of low bone density must put the emphasis on spony bone. PMID- 12561523 TI - [Detection, identification and investigation on small round structured virus in the fecal specimen of diarrhea patients]. AB - The indirect ELISA method with rabbit anti-SRSV superimmune serum on detecting small round structured virus(SRSV) in fecal specimen was established. The result was distinctly different in positive and negative controls, which demonstrated that the ELISA method established in this study was accurate and credible for detecting SRSV. Fecal specimens have been collected from 45 diarrhea patients in Beijing during the autumn and winter in 1998-1999. One specimen showed strong SRSV-positive result and two specimens showed weak SRSV-positive results. Furthermore, SRSV-like virion could be observed in the specimen with strong SRSV positive result under electronic microscope with phosphate tungsten negative stain and 60000-fold magnification. But no typical SRSV-like virion could be observed in specimens with weak SRSV-positive results. The rate for SRSV infection in diarrhea patients observed in this study was about 2.2%. The results implicated that SRSV infection was sporadic in Beijing during the autumn and winter seasons in 1998-1999. PMID- 12561524 TI - [Investigation and evaluation on the harmfulness of partial vibration and the contact vibration time]. AB - The harmfulness from partial vibration with extended contact vibration time was studied by investigating into the symptoms and signs of workers with local vibration disease in Taiyuan Iron & Steel Company. The sense of hand, the function of nerve ending and the microcirculation of fingernail wrinkles of 321 workers working on local vibration operations were examined and observed. The indexes examined are worse with the increase of contact time and are better in low contact group. A positive correlation between the abnormal of microcirculation, the pain and numbness of hand was observed. It is concluded that the degree of harmfulness on hands and upper limbs of whom working on local vibration operation is related to the time of contact vibration. The function of nerve ending and the ending circulation were injured at the same time. PMID- 12561525 TI - [Studies on the genotoxic effects of acrylonitrile]. AB - The effect of acrylonitrile (AN) on the reproduction was investigated in 341 male and 365 female exposed workers, 384 male and 438 female unexposed controls by questionnaires on the process of childbearing and the outcome of child birth. The results showed that AN caused menstrual disorder and dysgenesis in exposed female workers and the wife of exposed male workers. It is concluded that AN is harmful to the reproduction of exposed male and female workers. PMID- 12561526 TI - [Effects of air pollution from burning coal on respiratory diseases in adults]. AB - A cross sectional epidemiological study on the effect of air pollution from burning coal on the occurrence of respiratory symptoms and diseases in adults was carried out in Taiyuan city of Shanxi province. Adult residents above 25 year-old living in three areas with different degrees of pollution were selected. The results showed that the prevalence of respiratory symptoms and diseases increased with the extent of air pollution after age, sex, occupation, smoking, indoor coal burning and history of diseases were adjusted with Logistic model. PMID- 12561527 TI - [Optimizing the synthesis of hydroxyapatite as a method for the defluoridation of drinking water]. AB - As the cheaper raw materials for the synthesis of hydroxyapatite as a defluoridator, calcium hydroxide and phosphoric acid are used. Hydroxyapatite is made by a hydro-synthesis method, and optimum conditions are determined by batch test for defluoridation with respect to the consumption of phosphoric acid, reaction time and acid-washed product. The results showed that the defluoridation capacity of the synthetic hydroxyapatite for break through (F- 1.0 mg/L is considered as the break through point) was F- 5.6 mg/g in column test, and F- 10.7 mg/g for saturation in column test. After optimization of the synthesis method, the defluoridation capacity of hydroxyapatite is much higher than activated alumina and bone charcoal, which are used currently. PMID- 12561528 TI - [Determination of domoic acid in water and aquatic animals by high performance liquid chromatography]. AB - The content of domoic acid in water and in the body of some aquatic animals was detected by HPLC in order to find out the pollutant condition. The result showed that HPLC analysis was a simple and effective method for determining domoic acid in water and aquatic animals. Domoic acid has been found in some marine shellfish and not in seawater or freshwater. It is necessary to strengthen monitoring the contamination of domoic acid to prevent the toxin poisoning. PMID- 12561529 TI - [Prospects on the application of toxicogenomics in assessing the safety of chemical compounds]. AB - Toxicogenomics has been developing with the rapid advances in Human Genome Project in recent two years. The categories and specialty of tox-gene; the application of tox-gene microarray in safety assessment, analysis of gene mutation and gene polymorphism, extrapolation of animal tests results, clarification of mechanism and safety assessment of mixtures; some problems of toxicogenomics need to be solved in spite of its great prospective application. PMID- 12561530 TI - [Characters of uncoupling protein and its relation with obesity]. AB - Uncoupling proteins(UCP) are carrier proteins in mitochondria. In eukaryotic cells, ATP is generated by oxidative phosphorylation, an energetic coupling at mitochondria level. The oxidative reactions occurring in the respiratory chain generate an electrochemical proton gradient at both sides of the inner membrane of mitochondria. This gradient is used by the ATP synthase to phosphorylate ADP into ATP. The coupling of cell respiration with ADP phosphorylation is only partial in brown adipose tissue (BAT) mitochondria, where UCP causes a reentry of protons into the matrix and abolishes the electrochemical proton gradient. The liberated energy is then dissipated as heat and the synthesis of ATP is reduced. Recently, the cloning of new UCPs expressed in other tissues revealed the importance of this kind of regulation of respiratory control in metabolism and energy expenditure. The newly characterized UCPs are potential target drugs for obesity treatment, which could be favor of energy expenditure and diminish the metabolic efficiency. PMID- 12561531 TI - [Toxicity of manganese exposure on the postnatal development of brain in mice]. AB - The toxicity of manganese exposure on the postnatal development of brain in mice was investigated. Muscle motor ability was examined by Morris Water Maze, spontaneous action by Open Field Maze, neuro-anxiety by Elevated Plus Arms Maze. Lipid peroxides (LPO) level and superoxide dismutase (SOD) activity of brain was detected by neurobiochemical assay; glial fibrillary protein acid (GFAP) immunoreactivity was observed by cytoimmunochemistry. The results showed that in Mn-exposed group: (1) the length of body was significantly reduced; (2) the latency for reaching the platform above water in Morris Water Maze Test was longer, the walking squares and spontaneous rears in Open Field Maze, and the entries to open arms and the ratio of entries for open arms to entries for total arms in Elevated Plus Arms Maze were significantly increased; (3) LPO was obvious increased and SOD activity was decreased in brain; (4) GFAP immuno-reactivity and the average proportional density of GFAP-positive products in dendritic gyrus of cerebral cortex were significantly increased. More significant changes had been observed in the high Mn-exposed group. It was suggested that manganese exposure could lead to behavioral, neurobiochemical and cytoimmunochemical toxicity of brain in postnatal development. PMID- 12561532 TI - [Analysis on the components of rat urinary stone induced by terephthalic acid]. AB - In order to analyze the components of rat urinary stone and to explore the possible mechanism of chemically induced bladder cancer, terephthalic acid (TPA) is orally administrated to rats at the doses of 5000 and 500 mg/kg/BW/everyday for consecutive 90 days and stones in bladder and kidney are collected. Inductive Coupled Plasma Quantomer (ICP), Element Analyzer (EA) and Fourier Transform Infared Spectrometer (FT-IR) are applied for the analysis on the components of stone in bladder and kidney. The results showed that the main components of the stone are calcium, carbon, nitrogen, hydrogen and phosphorus. The FT-IR showed that the stones both in bladder and kidney might contain nitrous, carbonate, ammonium salt, and para-position replaced benzene-ring compounds, but no original TPA is detected. The findings indicate that TPA might have a metabolic turnover inside the rat body, and, at least, might not be completely excreted in its original form. PMID- 12561533 TI - [Study on lipid peroxidation of electrolyzing-aluminum workers]. AB - In order to study the changes in lipid peroxidation level and anti-oxidase activity in electrolyzing-aluminum workers exposed to both aluminum and fluoride and to find out the sensitive biological monitoring indicators, 65 electrolyzing aluminum workers are recruited as exposed group and 52 healthy workers as control group. Serum and urine aluminum and fluoride concentration, serum glutathione peroxidase(GSH-Px) activity, serum malonyl dialdehyde (MDA) concentration, whole blood superoxide dismutase(SOD) activity, serum cuprum and zinc concentration, and X-ray of skeleton are detected and taken. The results showed that aluminum and fluoride concentration in serum and urine increase, GSH-Px activity reduces, MDA concentration increases and SOD activity increases, cuprum concentration increases and zinc concentration reduces significantly (P < 0.001) in exposed group compared with those of control group. Both serum aluminum and fluoride concentration are correlated inversely with GSH-Px activity, and correlated with MDA concentration and SOD activity. It is suggested that serum aluminum and fluoride concentration can clearly reflect the loading capacity of body aluminum and fluoride. Occupational exposure to both aluminum and fluoride lead to rising of lipid peroxidation and abnormal metabolism of cuprum and zinc before changes in skeleton occurred. Serum MDA concentration and GSH-Px activity might be the potential early-stage indicators of biological surveillance for electrolyzing aluminum workers' health. The increase of SOD activity may be a kind of body compensations. PMID- 12561534 TI - [Study on metabolism of preventive anti-fluorine agent on rats]. AB - In order to study the metabolism of preventive anti-fluorine agent, 30 rats are randomly divided into high-dose, low-dose and a control groups. The high dose (400 mg/kg.d) and low dose (16 mg/kg.d) are orally administrated respectively, and the content of boron and/or zinc in urine, dung, serum, bone, liver, muscles, brain tissues is determined. The results showed that during the administration of this agent, the content of boron in urine and the content of zinc in dung increases obviously in both high-dose and low-dose groups and their discharge rate is consistent with the dose given. The content of boron and zinc in bone, liver, and zinc in serum, muscles, brain tissues increases evidently compared to that in control group but decreases rapidly after administration of the agent. The findings revealed that there is a rapid metabolism of boron and zinc in the body of rats. The highest content of the agent is observed in bones. The content ranks second in liver and muscles but no accumulative effects are observed. PMID- 12561535 TI - [Study on the regeneration methods of the synthetic hydroxyapatite as a material for defluoridation of drinking water]. AB - The regeneration efficiencies of the synthetic hydroxyapatite saturated by fluoride using either sodium hydroxide method or surface coating method are studied and compared. Both static and dynamic conditions for surface coating regeneration method are also tested. The results showed that the regeneration efficiency of the synthetic hydroxyapatite saturated by fluoride treated by surface coating method is 46%-64%, more than two times of that of it treated by the traditional sodium hydroxide regeneration method. PMID- 12561536 TI - [Oxidative stress of cooking oil fume on rat type II lung cells]. AB - The relationship between the oxidative stress and damages in rat type II lung cells exposed to cooking oil fume (COF) was studied. The cytotoxicity, DNA cross links and DNA single strand breaks could be observed in the cells exposed to COF. The contents of MDA were increased and GSH were decreased significantly with exposure doses of COF and with time dependence. Pretreatment with antioxidant N acetylcysteine (NAC) could reduce the toxicity of COF to the cells. The results suggested that cytotoxicity and DNA damages of rat type II lung cells induced by cooking oil fume might be related to the oxidative stress and the possible pathways might be the formation of lipid peroxides and interfering the GSH anti oxidative systems of the cells. PMID- 12561537 TI - [Effects of chronic alcohol intake on serum glucose and insulin of rats]. AB - In order to study the effects of chronic alcohol intake on serum glucose, insulin and glucose tolerance test (GTT), a oral administration of alcohol solution (6 ml/kg.d) in different concentration (10%, 30% and 50%) is given to rats for 60 days. The results showed that the fast serum glucose of rats in the groups orally administrated with 30% and 50% alcohol solution increases obviously in comparison with that in control group. There exist abnormalities in 1 hour, 2 hour and, 3 hour GTT (P < 0.05) in these groups and the difference in gender in both groups is statistically significant. It is also shown that the level of serum insulin of rats decreased along with the increase of alcohol dosage and is significantly lower in the group of rats administered 50% alcohol solution than that in the normal control group (P < 0.05). These results demonstrated that chronic alcohol intake causes the increases of fast serum glucose and abnormality of glucose tolerance test (GTT), and likely, the final abnormality of glucose metabolism in which the decrease of insulin secretion might be one of the contributing factors to the pathogenesis. PMID- 12561538 TI - [Effect of zinc deficiency on the behavior and calcium status of hippocampal neurons in rats]. AB - In order to explore the effects of zinc deficiency on the behavior, [Ca2+]i and active CaM contents of hippocampal cells of rats, 21 male Wistar rats were randomly divided into 3 groups, i.e. zinc deficiency group (ZD), pair-fed group (PF), and control group (CT) fed with zinc deficiency (ZD group) and normal diets (PF and CT group). At the end of the third week, the behavior in open-field test was observed. Then all the rats were killed and [Ca2+]i concentration and active CaM contents of hippocampal cells were determined by Fura-2 two wave-length fluorophotometry and flow cytometry respectively. It showed a significant difference between ZD and CT group in behavior was observed. [Ca2+]i concentration of ZD group was remarkably higher than that of CT and PF group. Active CaM contents of ZD and PF groups were significantly lower than that of CT group, and the active CaM content of ZD was also significantly lower than that of PF group. It showed that the effect of zinc deficiency on behaviors may be related with the changes in calcium status of hippocampal cells in growing rats. PMID- 12561539 TI - [Effects of selenium-enriched garlic on blood lipids and lipid peroxidation in experimental hyperlipidemic rats]. AB - The modulating effects of selenium-enriched garlic on lipid metabolism disorder and lipid peroxidation were studied in hyperlipidemic rats induced by high fat diet. Adult male and female Wistar rats were divided into 6 groups: (A) normal control; (B) high fat diet (HFD) control; (C) HFD + selenite; (D) HFD + selenium enriched garlic; (E) HFD + common garlic; (F) HFD + common garlic + selenite. The selenium content of diets in groups A, B and E was 0.08 mg/kg diet, while that of the other 3 groups was 2.7 mg/kg diet. At the end of the experimental period (12 weeks), blood and liver were collected for biochemical measurements and for histopathological examination of liver. The results showed that the serum concentrations of TC, TG and LDL-C in groups C, D and F were significantly lower and HDL-C higher than group B. Female rats were more sensitive to HFD exposure than male rats. The peroxidative status of all four experimental groups was significant inhibited as shown by the lower lipid peroxide (MDA) in liver and higher activities of GPX in erythrocytes and liver and SOD in plasma. Selenium contents in liver and kidney of male rats in groups D and F were higher than group C. Significant accumulation of selenium in erythrocytes was observed in groups D and F. The liver of all four experimental groups revealed ameliorated fatty liver induced by HFD. The amelioration of group D was more prominent than other three experimental groups. The results suggested that selenium-enriched garlic is superior to selenite or common garlic in decreasing the blood lipid level and peroxidative status and slightly better than combined common garlic and selenite. PMID- 12561540 TI - [Study on animal models for hyperlipidemia]. AB - To establish a better animal model for hyperlipidemia, three animal models with SD rats, Wistar rats and golden hamsters were explored. The semi-purified diet for rats was composed of 1% cholesterol, 10% lard, 10% yolk powder and 79% normal diet. The semi-purified diet for hamster was composed of 0.2% cholesterol, 15% lard and 85% normal diet. Serum TC, TG, HDL-C, liver cholesterol and triglyceride were assayed. Serum TG of SD rat, Wistar rat and hamster was 194.4%, 86.2% and 84.3% respectively higher than that of control after 5 weeks of experiment. Serum TC of the three groups was 76.8%, 48.3% and 134.8% respectively higher than that of control. The results suggested that hamster might be a better animal model for hypercholesterolemia and SD rat might be better for hypertriglyceridemia. PMID- 12561541 TI - [Effect of serum leptin on nutritional status of patients with cancer]. AB - In order to investigate the effect of serum leptin and its potential as a parameter for the accessment of nutritional status of patients with cancer, serum leptin concentration, body mass index (BMI), blood erythrocyte, hemoglobin, serum albumin, lipid and lipoprotein concentration of 325 cancer patients and 66 healthy controls are meatured. The incidence of patients with BMI < 18.5, hypoalbumia, anemia, hypercholesterolemia, and hypertriglyceridemia is 23%, 14%, 42%, 17.2%, and 21.4% respectively. The incidence of patients with BMI < 18.5 in pulmonary carcinoma is obviously lower than that of those with gastric carcinoma (P = 0.022). The incidence of patients with hyperglycosemia and hypertriglyceridemia in pulmonary carcinoma is obviously higher than that of those with gastric carcinoma (P = 0.003 and P = 0.029 respectively). Serum leptin concentration in the patients with malnutrition is significantly lower than that of those with no malnutrition and that of those obese patients (P = 0.000). There is no significant difference in serum leptin concentration between patients with cancer and healthy control persons with same gender and with BMI value ranged from 18.5 to 25. It is shown that the BMI, gender and serum albumin concentration are all influencing factors to the serum leptin concentration and the serum leptin concentration is significantly correlated with BMI, gender and serum albumin concentrations (r = 0.599-0.698, P = 0.000). The above mentioned results from this study indicate that there is a high anemia incidence of patients with cancer. Serum leptin concentration can reflect the changes in BMI and nutritional status of the patients with cancer. The serum leptin concentration has the potential of being a parameter for assessing nutritional status of the patients with cancer. PMID- 12561542 TI - [Cloning of ZnT3 cDNA and expression of ZnT3 mRNA in mice]. AB - In order to determine the level of mouse zinc transporter3 (ZnT3) mRNA expression, the fragment is cloned by RT-PCR method and is sequenced by dideoxynucleotide method. The expression of ZnT3 mRNA is also examined by RT-PCR method and is compared with beta-actin that served as the control. As the result, a proper single fragment of 700 bp is obtained with the sequence conformable to the corresponding fragment sequence of reported mouse ZnT3 cDNA. The ZnT3 mRNA is detected in cerebrum, hippocampus and in testis in that the highest expression is observed. However, the expression of ZnT3 mRNA is not detected in heart, liver, lung, spleen, kidney, intestine, olfactory bulb and cerebellum. The expression of ZnT3 mRNA in brain and testis suggests a physiologically functional importance of ZnT3 in mental and reproductive activities of mouse. PMID- 12561543 TI - [Inhibitory effects of genistein on the synthesis of DNA and the protein expression of cyclin D1 in human gastric carcinoma cell-line]. AB - In order to study the inhibitory effects of genistein on the growth of human gastric carcinoma cells and to explore its possible mechanism, cell proliferation and DNA incorporation tests are applied. Immunocytochemistry is also applied in this study to detect the protein expression of cyclin D1 after the cell-line is exposed to genistein. The results showed that the growth of the cells and the synthesis of DNA is decreased in a marked dose-dependent manner after the treatment with genistein. Genistein also decreased the protein expression of cyclin D1. The above mentioned results indicate that genistein inhibits the growth of gastric carcinoma cells and the possible mechanism of this inhibition might be resulted from inhibiting the synthesis of DNA and the protein expression of cyclin D1. PMID- 12561544 TI - [Production and characterization of monoclonal antibody against the O antigen of Enterohemorrhagic Escherichia coli]. AB - Spleen cells collected from Balb/c mice immunized with the somatic antigen of Enterohaemorrhagic E. coli O157:H7 are fused with murine Sp2/0 myeloma cells. A hybridoma cell line specific to E. coli O157 is established after screening and sub-cloning and is designated as 3A5. It belongs to a subtype of IgM and the ascetic titer of the antibody is 1:1 x 10(6). The detection limit is 10(5)-10(6) cfu/ml. This monoclonal antibody(McAb) also has activity with Enterohaemorrhagic E. coli O113:H21. PMID- 12561545 TI - [Effects of sterigmatocystin on interleukin-2 secretion of human peripheral blood mononuclear cells in vitro]. AB - Sterigmatocystin (ST) is one of predominant contaminating mycotoxins in foodstuffs and grains of high incidence areas of malignant tumors in China. The effect of ST on interleukin-2 (IL-2) secretion of human peripheral blood mononuclear cells (HPBMc) in vitro was determined with enzyme-linked immunosorbent assay (ELISA) method to explore its putative effects on human immune function. ELISA analysis revealed that ST treatment generally showed negative effects on the IL-2 secretion of HPBMc in vitro. As the ST concentration changes, the inhibiting effects were different. The inhibiting effects at low concentrations (0.03125-0.125 mg/L) and high concentrations (1-2 mg/L) were stronger than the other concentrations(P < 0.05). The time-effect analysis (ST 1 mg/L) showed that inhibiting effects of ST on IL-2 secretion of HPBMc could be seen to a variable degree from 1 to 64 h after ST treatment, while a significant time-effect correlation could be found from 8 to 64 h (r = 0.822, P < 0.05). The results obtained in present study showed that ST has certain inhibiting effects on the secretion of IL-2 of HPBMc in vitro. PMID- 12561546 TI - [Effect of grape seed extracts on blood lipids in rabbits model with hyperlipidemia]. AB - The effect of grape seed extracts on serum lipids was observed in rabbits fed with high fat diet. The control group fed with normal stock diet. The diet of the four experimental groups was high in fat and cholesterol. The experimental control group fed with the high fat diet only, and the other three experimental groups fed the high fat diet added with grape seed extracts (5, 10 or 15 g/d) for three months. At the 0th, 6th and 12th week of the experiment, there was a lower serum TC, TG, LDL-C and a higher HDL-C in experimental groups in comparison with experimental control group. PMID- 12561547 TI - [Effect of excessive Zn2+ and deficient Zn2+ on the expression of wild type-p53 mRNA in rat glicoma cells]. AB - RT-PCR is applied to observe the effects of excessive and deficient Zn2+ on the expression of wild type-p53 (wt-p53) in rat glicoma cells. Rat glicoma cells in garithmic growth phase is treated with 0.3 mmol/L Zn2+ and 5 mumol/L TPEN respectively for 24 hours. Results show that the expression of wt-p53 of rat glicoma cells increased obviously in the deficient zinc group but does not increased in the excessive zinc group. These indicate that the expression of wt p53 mRNA of rat glicoma cells is up-regulated by deficient zinc and inhibited by excessive zinc. PMID- 12561548 TI - [Study on the composition of Lycium barbarum polysaccharides and its effects on the growth of weanling mice]. AB - In order to observe the effects of lycium barbarum polysaccharides (LBP-4) on the growth of weanling mice and the absorption of some metals in the their body, the composition of LBP-4 is determined. 120 female weanling mice are divided in random into 4 groups. They are fed on LBP-4 at the dose of 5, 10 and 20 mg/(kg.d) respectively. The taken feed weight and the body weight of mice are recorded everyday. After 21 days, the content of calcium, magnesium, zinc and iron in pygal muscles and femora of mice is determined. The results showed that LBP-4 is composed of six kinds of monosaccharides that can enhance food conversion rate and the content of zinc and iron in body of mice, and reduce the body weight. PMID- 12561549 TI - [Effects of refined konjac meal on lipid metabolism and blood viscosity of rats fed by high fat forage]. AB - Male SD rats are fed by high fat diet supplemented with refined konjac meal for 6 weeks and the effect of refined konjac meal on the serum lipid peroxides (LPO) and blood viscosity are observed. The results showed that the refined konjac meal can obviously decrease serum cholesterol, triglyceride and serum LPO of rats in comparison with those of rats fed only by high fat forage, and can elevate, at the same time, the high density lipoprotein-cholesterol and high density lipoprotein-cholesterol/triglyceride value. It is also shown that the refined konjac meal can decrease the blood viscosity, but has no effect on forage intake and weight gain of rats. PMID- 12561550 TI - [A simple method for screening of highly inhibitory Lactic acid bacteria on Aspergillus flavus growth]. AB - A simple and rapid method for screening of Lactic acid bacteria with high inhibitory power is developed. The methodology employed was the agar-diffusion assay, which was standardized for medium composition, and incubation conditions. The assay was composed of two layers of medium, MRS agar at the bottom of plate, PDA at the top of MRS agar. Lactic acid bacteria culture was inoculated to the media between MRS and PDA, then a known number of mold spores was surface plated on PDA plates, and incubated at 25 degrees C for 72 h. PMID- 12561551 TI - [Determination of chloroform and tetrachloro-methane in water with capillary column-microECD]. AB - The method of determination of chloroform and tetrachloro-methane in water with capillary column-microECD had better and wider linear range and lower detection limit. At the range of 0-10 micrograms/L chloroform and 0-1 microgram/L tetrachloro-methane, the regression equation were calculated as Y = 1.959X + 4.816 (r = 0.9910) and Y = 3.947X + 42.58 (r = 0.9970). The lowest detection limit were 0.1032 microgram/L chloroform and 0.01219 microgram/L tetrachloro methane. This method not only simplified the operating processes, but also increase the sensitivity, and reavled better precision and accuracy. PMID- 12561552 TI - [A new measurement for assessing the intra-household food distribution]. AB - Discrepancy Score and Food Share to Energy Share were used to describe food distribution within a family. The results showed that dietary and nutritional status of people with discrepancy score below zero was worse than that of people with discrepancy score above zero. It proved that food allocation was closely related to dietary and nutritional status. PMID- 12561553 TI - [Individual susceptibility to hematotoxicity from benzene exposure and the genetic polymorphism of metabolic enzymes]. AB - Benzene is one of wildly used chemicals. Long-term exposure to benzene causes hematotoxicities and further, the development of including anemia, myelodysplastic syndrome (MDS), aplastic anemia, etc., with the leukemia as the worst. People vary greatly in their susceptibility to adverse health outcomes from benzene exposure. The author reviewed the relationship between genetic polymorphism of I metabolic enzymes(CYP2E1, NQO1, MPO) and II metabolic enzymes(GST, PST) involving benzene metabolite and interindividual variation in their genetic susceptibility to hematotoxicity from benzene exposure in this paper. PMID- 12561554 TI - [Analytical methods used in pharmacokinetics studies for biotechnology drugs]. AB - In order to provide a reference for selecting analytical methods in pharmacokinetics studies for biotechnology drugs such as recombinant proteins, monoclonal antibodies or genetic engineering antibodies and oligonucleotide drugs, the following methods such as radiolabels, immunoassays, bioassays, chromatography and even the novel protein-chip technique are reviewed. Their mechanisms, application fields, and advantages and limitations are also summarized. PMID- 12561555 TI - [Manufacture of hydroxyapatite as a defluoridator and the mechanism of defluoridation for drinking water]. AB - Hydroxyapatite as a defluoridator for drinking water is characterized by a large capacity of defluoridation, and easy and simple operation in application. The quality of drinking water is not changed, the sorbent is easy to be regenerated, and the second pollution does not occur after the treatment of drinking water with the sorbent. Hydroxyapatite method has advantage over the methods such as actived alumina, bone char and electrodialysis commonly used in defluorination of drinking water. It is for this reason that the components, structural feature, synthesis and application of hydroxyapatite and its mechanism of defluoridation are summarized in this paper. PMID- 12561557 TI - [Application of single cell gelelectrophoresis assay in environmental hygiene]. AB - The single cell gelelctrophoresis assay is a rapid, simple and sensitive technique for measuring DNA damage and repair at the single cell level. It is introduced into environmental hygiene and plays an important role in many aspects such as detecting and evaluating the genotoxity of environmental toxic substances. In this paper, the underlying principles of this assay in detecting DNA damage and repair, and its application in environmental hygiene are reviewed. PMID- 12561556 TI - [Molecular mechanisms of environmental estrogens on reproductive and developmental toxicity]. AB - It has been shown that numerous natural and synthetic chemicals with estrogenic activities are reproductive or developmental toxic factors in humans or animals by epidemiological investigations and animal experiments. The mechanisms of toxicity, however, are very complicated, especially the molecular mechanisms. This article introduced the current progress on molecular mechanisms of toxicity, which included modulating signaling pathways, interfering gene expression during embryo stage, inducing mutation, etc. PMID- 12561558 TI - [Application of dietary intake references in the diet assessment]. PMID- 12561559 TI - [Effects of low-level lead exposure on neurobehaviour development in 1-3 year-old children and the intervention guideline]. AB - In order to study the effects of low-level lead exposure on neurobehaviour development, the neurobehaviour functions in eighty-five 1-3 year-old children were assessed by Neurobehaviour Psychological Test (recommended by the Institute of Psychology, Chinese Academy of Sciences). The results showed that the increase of blood lead (BPb) level in each 100 micrograms/L was associated with an average loss of 3-4 points of Development Quotient (DQ) in these subjects. Age, hand mouth habits and environmental lead pollution are the main risk factors. It is concluded that even in a low-level lead exposure, DQ in young children may be affected. Certain intervention steps based on the levels of lead exposure were recommended. PMID- 12561560 TI - [Effect of methylmercuric chloride on apoptosis and c-fos expression of brain glial cells]. AB - In order to explore the injury of brain development caused by methylmercuric chloride(MMC) and the relationship between the injury and c-fos expression, the damaging effects of MMC on the morphology of cultured brain glial cells derived from neonatal Wistar rat were studied by microscope and electronic microscope, and the effect of MMC on the c-fos expression of glial cells were studied by immunocytochemical method. The results showed that the characteristics of apoptosis, such as condensed cytoplasm, marginal nuclear chromatin and pyknotic nuclei were found after cultured glial cells was exposed to MMC. The expressed c Fos protein began to increase in the nuclei of cultured glial cells at 0.5 h after MMC was added to DMEM medium for 10 min and increased markedly with time prolonged. The percentage of Fos-positive nuclei reached a peak at 2 h and declined gradually from 4 to 6 h. The percentage of Fos-positive glial nuclei in the group exposed to 0.32 mmol/L MMC was significantly higher than those in other groups, which were exposed to 0.00125, 0.005 and 0.02 mmol/L MMC respectively (P < 0.01) at 2 h after MMC was added. It is concluded that MMC can induce the expression of c-fos and the apoptosis of glial cells in vitro. The findings suggested that the apoptosis might be mediated by c-fos expression. PMID- 12561561 TI - [Study on the bladder calculi and bladder cancer induced by terephthalic acid in rats]. AB - The carcinogenicity and calculogenesis of terephthalic acid on bladder was investigated. Terephthalic acid at doses of 5000, 500 and 50 mg/kg body weight/d were administrated to SD rats for 90 days consecutively. Control animals received normal feeds. Bladder calculi were induced in 5000 mg/kg (10/17) and 500 mg/kg (2/18) group. Calculi incidence of male rats was higher than that of female rats in 5000 mg/kg dose group. Bladder transitional cell cancer (4/17), atypical hyperplasia (5/17) and single hyperplasia (7/17) could be observed in 5000 mg/kg group. Atypical hyperplasia (10/18) and single hyperplasia (5/18) could be observed in 500 mg/kg group. Single hyperplasia (10/17) could be observed in 50 mg/kg group. The controls were normal except 1 case of single hyperplasia (1/18). There was a positive correlation between pathologic changes and dosage, but no significant correlation between pathologic changes and calculi incidence. Those rats without calculi produced some changes too, but no squamous metaplasia and carcinoma happened. The results suggested that under the study conditions, squamous cell carcinogenicity might be induced by terephthalic acid, and the calculi might not be the major factor inducing cancer. PMID- 12561562 TI - [Study on the injury of liver induced by terephthalic acid ethylene glycol and/or dowtherm A in rats]. AB - The joint injury actions and mechanisms of terephthalic acid (TPA), ethylene glycol (EG) and/or dowtherm A (DOW) on liver in rats were investigated. A subchronic toxicity study was designed by a 2(3) factorial method. Some enzymes, biochemical and morphologic indices reflecting the injury of liver were studied. The results showed that serum ALT and serum total bile acid (TBA) of rats in the combined intoxication groups were significantly higher than those in the groups with single toxic agent and control group. The results of factorial analysis showed that the joint action induced by TPA, EG and/or DOW were characterized as additive (TPA + EG), synergistic (EG + DOW), synergistic (TPA + DOW) and additive(TPA + EG + DOW) actions. The deduction was identified by morphologic observations. PMID- 12561563 TI - [Effects of chlorine on the antigenicity and RNA polymorphism of hepatitis A virus]. AB - The mechanisms of HAV inactivated by chlorine was investigated by enzyme-linked immunosorbent assay and RNA fingerprinting. Arbitrary primed PCR were involved to detect the changes of antigenicity and RNA polymorphism respectively before and after inactivation by chlorine. The results showed that the infectivity was completely inactivated at 10 mg/L or 20 mg/L of chlorine for 30 minutes and the contacting time should be prolonged to 60 minutes when the loss of antigenicity was observed at the same condition. The changes of RNA polymorphism were often observed while the infectivity was completely inactivated except the concentration of chlorine was 10 mg/L for 30 minutes. The results implied that HAV inactivation by chlorine might be the result of destruction of some fractions of HAV genome but not the whole HAV genome. PMID- 12561564 TI - [Study on the denitrification of drinking water with upflow anaerobic sludge blanket reactor]. AB - The characteristics of denitrification was investigated with a pilot scale upflow anaerobic sludge blanket (UASB) reactor at room temperature. The results showed that when brewery waste degrading sludge was used as seed, the starting process was completed within 7 weeks, with hydraulic residence time shortened from 11.1 h to 4.7 h, COD/N/P = 200/5/1 and influent NO3-(-)N concentration increased from 5 mg/L to 100 mg/L. After the process starting, the most probable number [n(MPN)] of denitrifying bacteria was 60 folds and the maximum velocity of CH4 produced was 10 folds higher than before. The removal efficiency of No3-(-)N was 99%, C/N ratio and pH value were investigated as effect factors. When C/N > = or 1.0, the NO3(-)-N removal efficiencies were not different from those of C/N < 1.0 groups significantly. The pH value could meet the discharge standards. PMID- 12561565 TI - [Experimental study on the potential carcinogenicity of cooking oil fume condensate]. AB - The potential carcinogenicity of cooking oil fume condensate (COFC) to human was studied. Human embryo lung diploid fibroblast cell strain KMB-17 cell was applied to establish a human diploid cell transformation system in vitro. Different concentrations of COFC were added into the media and co-incubated with cells. The malignant degree of transformation was assessed by the biological characteristics of the cells. The concentrations of COFC within the dose range of the experiment could induce the malignant transformation of KMB-17 cell, and with a obvious dose response relationship (r = 0.9811). Transformed cells have exhibited many characteristics associating with malignant transformation, such as loss of density and contact-dependent inhibition, growth at low serum concentration, agglutination by low concentration of Con A, alteration of karyotype from diploid to aneuploid, and lose of anchorage dependence. It suggested that the malignant transformation of human embryo lung diploid fibroblast cell strain KMB-17 cell could be induced by COFC, which might have potential carcinogenicity to human. PMID- 12561566 TI - [Effect of Jingtian compound on the delay of skin aging]. AB - In order to study whether Jingtian compound extracted from herbs could delay the process of skin aging, the skin on the back of Guinea pigs (6 and 15 months old) were shaved topically and applied with and without 0.5% Jingtian compound for 30 days by self-control design. The possible alterations caused by Jingtian compound were observed by histological and biochemical techniques. Results showed that the number and the activity of fibroblast in dermis was increased prominently compared with the control. The activities of superoxide dismutase, glutathion peroxidase and the hydroxyproline level of acid soluble collagen in dermis were enhanced, and the malondialdehyde content was inhibited concomitantly. It is suggested that Jingtian compound might play a protective role on skin aging. PMID- 12561567 TI - [Anti-lipoperoxidation of grape seed extracts in vitro]. AB - The anti-lipoperoxidation of grape seed extracts (GSE) was observed by malondialdehyde (MDA) generated in liver and brain homogeneates spontaneously or induced by CCl4, H2O2, Fe(2+)-ascorbic acid. The exhaustion of glutathione (GSH) in liver homogenate initiated by Fe(2+)-ascorbic acid was also tested. Results showed that GSE significantly inhibited the generation of MDA in rat liver and brain homogenates both spontaneously formed or induced by CCl4, H2O2 and Fe(2+) ascorbic acid with dose-effect relationships. GSE could also reduce the exhaustion of GSH levels in mice liver. The study indicated that GSE is effective on the inhibition of lipid peroxide and on the protection of liver from injury caused by lipid peroxidation. PMID- 12561568 TI - [Effect of lycium barbarum L on defending free radicals of mice caused by hypoxia]. AB - The effect of Lycium Barbarum L. (LB) on the free radical defending system during hypoxia was tested. Fifty six mice were divided into 3 groups. Experimental mice were fed with LB and the control mice were fed with distilled water by tube feeding for 16 days. Animals were put into each enclosed bottle for testing the effect of hypoxia. The results showed that LB treatment could not prolong the survival time (P > 0.05), but increase the activities of superoxide dismutase (SOD), catalase(CAT) and total anti-oxidative capacity as compared with the control group(P < 0.05). The study indicated that LB could not increase the hypoxic tolerance, but it might have a protective effect on free radical injury caused by hypoxia. PMID- 12561569 TI - [Inhibition on the endocytosis of early visceral yolk sac induced by homocysteine in rats]. AB - The effect of homocysteine (HCY) on the development and endocytosis of rat visceral yolk sac (VYS) was investigated. A preliminary method on early (GD9.5) rat yolk-sac placenta cultured in vitro was established by techniques using microdissection and transplantation. The results showed that HCY did damage VYS development and blood vessel formation, the damages were characterized by shrunk surface, small or defective blood island and significant reduction of diameter (P < 0.01). HCY also inhibited the process of VYS transport, specifically the endocytosis (as measured by [U-14C] sucrose uptake), the rate of endocytosis was significantly decreased when HCY was included in the medium (P < 0.01). These results suggested that the inhibition of sanguimotor development and the reduction of endocytosis induced by HCY may directly correlate with the teratologic and embryopathologic events. PMID- 12561570 TI - [Study on the food consumption pattern of elderly people in four "cuisine" areas of China]. AB - In order to investigate the situation of food intake, diet habit and dietary pattern in people living in different "cuisine" areas of China, and to search for the relations between diet and health status, a total of 538 subjects (269 couples) aged between 50-60 years in four rural areas were surveyed by a food frequency questionnaire method. Selected items of food consumption pattern and food intake of the subjects were analyzed. The results showed significant differences on food intake among the people in the four cuisine areas. This study provided references for the future study on the relationship between diet and health. PMID- 12561571 TI - [Study on the identification of genomic modified foods]. AB - Nucleotide-based amplification method is an important system for the identification of genomic modified foods (GMF). Roundup Ready Soybeans (Monsanto company), Bt 176 GM maize (Novartis/Ciba-Geigy company) and Cecropin D capsicum was used as material to search for the feasibility of investigating the safety of GMF by PCR method. Primers specific for inserted genes and crop endogenous genes in Roundup Ready Soybeans, Bt 176 maize and Cecropin D capsicum were applied. The discrimination system for GM soybeans, GM maize and GM capsicum from the counterpart of non-GM products and the detection system for correlating marker genes and transgenes are established. The method was easy and fast, and the corresponding results fixed the standard or declared data. PMID- 12561572 TI - [Effects of dietary calcium-deficiency on bone development in growing rats and the mechanism involved]. AB - The effects of calcium on bone development in growing rats were studied by bone histomorphometry and the hormone changes in a model of calcium-deficient rats. Thirty growing Wistar rats were randomly allocated into three groups fed with low calcium diet, calcium-sufficient diet at libitum or pair fed with low calcium diet for 8 weeks. The calcium-deficient rats showed low bone mass, significantly decreased bone density and mineral content. The disorder of bone molding and ultrastructure at the femoral distal diaphysis of the calcium deficient rats were showed by histomorphometric parameters. The concentration of serum parathyroid hormone was significantly higher, calcitonin was significantly lower, and the content of hydroxyproline in bone was significantly lower. The results indicated that dietary calcium-deficiency might stimulate the secretion of parathyroid hormone, decrease bone mineral content and inhibit the synthesis of bone matrix, which might induce a defect on bone mineralization. PMID- 12561573 TI - [Development of a rapid separating incubation method for detection of Escherichia coli O157:H7 in food]. AB - To detect Escherichia coli O157:H7 in food, a selective and differential medium, CTV-MUG-RSMAC agar, was designed. Using this medium, the growth of most interfering organisms could be inhibited, and three main biochemical characteristics, including sorbitol-negative, rahmnose-negative and MUG-negative, to identify E. coli O157:H7 could also be simultaneously recognized. Sorbitol positive aberrance strains of E. coli O157:H7 could be distinguished on the medium. The procedure of detecting food sample includes adding sample to selective broth and incubating overnight at 42 degrees C, streaking the inoculum to CTV-MUG-RSMAC agar and incubating at 37 degrees C for 24 h, and picking the typical colonies to react with sera for O157 and H7 antigens, and then take biochemical tests. The procedure could be finished within 3 days. This method has been applied to 126 food samples, and E. coli O157:H7 was detected from 3 samples. The results were in coincidence with those using FDA method. PMID- 12561574 TI - [Inhibition of Lactobacillus species on the germination of Aspergillus flavus spore]. AB - Actively growing Lactobacillus plantarum CGMCC 1.1856 cells totally inhibited the germination of mold spores. Cell-free supernatant broth from the fermentation of Lactobacillus plantarum could not destroy the viability of mold spore. While the pH of the culture broth and supernatant were about 4.0, and the acidification of non-fermented broth to pH 4.0 with lactic acid could not cause a similar inhibition on spore germination. This experiment discounted the possibility that the inhibitory effect on mold growth was due to the lactic acid produced by Lactobacillus, and suggested that the effect was because of both low pH and microbial competition. PMID- 12561575 TI - [Expression of fused protein A-green fluorescent protein (PA-GFP)]. AB - The green fluorescent protein (GFP) of jellyfish victoria is an unusual protein with strong visible fluorescence both in vivo and in vitro. In this study, about 750 bp fragment of gfp gene was amplified by PCR and was inserted into pRIT2T vector containing protein A gene. The recombinant plasmid was transferred into E. coli JM101. GFP and protein A were fused and expressed in E. coli Top10. The bright green fluorescence on the plates could be visible under UV at 365 nm. The bacteria cells were disrupted by sonic oscillation. The fused protein PA-GFP could be obtained from the supernatants. PMID- 12561576 TI - [Determination of ochratoxin A by ELISA I. Study on the preparation of ochratoxin a antigen]. AB - Ochratoxin A (OTA) antigen was prepared by activated ester method. Factors influencing OTA antigen were discussed and the optimum conditions was found by L9(3(4)) orthogonal design. The results showed that when the mole ratio was OTA:BSA = 20:1, OTA:NHS(N-hydroxy-succinamide):DCC(dicyclohexylcarbodiimide) = 1:2:4, the activate time was 120 min and the conjugation time was 90 min, the utilization of OTA could reach 48.2% and the best conjugation ratio of OTA and BSA was 9.64. PMID- 12561577 TI - [Study on the absorption of quercetin and rutin at different segments of intestine]. AB - The absorption of quercetin and rutin at different segments of intestine was examined. Seventy two Wistar rats were divided randomly into 3 groups (control group, quercetin group and rutin group), each group was divided into 4 subgroups (duodenum group, jejunum group, ileum group and colon group). Different solutions (1% dimethyl sulfoxide, 20 mumol/L of quercetin and rutin) were perfused respectively. At the end of perfusion, effluent solution samples were analyzed with HPLC. The results showed that both quercetin and rutin could be absorbed at different segments of intestine, but the absorption of rutin was significantly lower than quercetin. On the length basis, the absorption rate of quercetin was colon > ileum > duodenum > jejunum. On the area basis, the absorption rate was ileum > jejunum > duodenum > colon. After the perfusion of quercetin, there were unidentified peaks appearing on HPLC chromatogram. The results suggested that quercetin was derived from intestine and then re-excreted into intestinal lumen. PMID- 12561578 TI - [Dietary fat on the procreation of male rats induced by dimethoate]. AB - The effect of dietary fat on the toxicity of dimethoate for the procreation of male rats was observed. Thirty two male rats were divided into 4 groups. 3 groups treated with dimethoate 12.5 mg/(kg.d) for 60 days were fed with high, moderate or low dietary fat respectively. The rats in control group were fed with water and moderate dietary fat. The results showed that serum testosterone(To) and LH and the activity of acid phosphatase and lactate dehydrogenase in testicle were higher in the high or low-fat diet groups than those in the moderate-fat diet group. It suggested that the highor low-fat diet could reduce the degree of harm in the testicle tissue of rats induced by dimethoate than that of moderate-fat diet. PMID- 12561579 TI - [Determination of serum type III procollagen in patients with rheumatoid arthritis and interstitial lung disease]. AB - Type III serum procollagen (Pc III) was measured by radio-immunossay in 30 rheumatoid arthritis(RA) patients with or without intestitial lung disease(ILD) and in 15 normal control subjects. The results showed that Pc III of RA patients were significantly higher than that of normal controls. Pc III levels is higher in RA patients with ILD than that in RA patients without ILD. The results suggested that RA patients with higher serum Pc III is more vulnerable to ILD, and the determination of Pc III might be valuable to show the complicated ILD. PMID- 12561580 TI - [Effect of tea polypheonls and ascorbic acid on the activity of antioxidant enzymes in rats with experimental silicosis]. AB - To explore the effect of tea polypheonls (TP) combined with ascorbic acid on the activity of antioxidant enzymes of rats induced by silica, the activities of nitric oxide synthase (NOS), glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) were determined. Compared with the normal saline control group, the activities of SOD and NOS of the silica-induced group increased (P < 0.05), but the activity of GSH-PX was gradually decreased. The activities of SOD and NOS decreased and GSH-PX increased in the intervention group. It is concluded that TP combined with ascorbic acid can alleviate the abnormal change of antioxidant enzymes in rats induced by silica. PMID- 12561581 TI - [Effect of taurine on blood and liver lipids in rats]. AB - The effect of taurine on serum and liver cholesterol level in rats and its mechanism was investigated. Female weanling SD strain rats (80-100 g) were divided into four groups, eight rats in each group. Group A fed with normal diet. The diet of groups B, C and D was normal diet incorporated with taurine at the levels of 6, 13.5 and 30 g/kg diet respectively. The experiment lasted for five weeks. Results showed that the blood cholesterol levels of rats with diets containing taurine were decreased. The decrease of plasma total cholesterol(TC), low density lipoprotein-cholesterol in the 6 g/kg taurine group was the most significant. Plasma triglycerides and apolipoprotein(Apo)-B in the 13.5 g/kg taurine group was decreased, and the amount of fecal bile acids was increased in this group. Plasma Apo-AI of the 30 g/kg taurine group was increased and the liver TC was decreased. PMID- 12561582 TI - [Evaluation and analysis on the micro-determination of fluoride in serum and finger blood]. AB - The relationship of fluoride concentration between blood serum from vein and blood from finger tip was explored. Ninety people exposed to different levels of fluoride were selected. Group A was exposed directly to high level of fluoride, group B was exposed indirectly to median level of fluoride, group C was exposed to low level of fluoride and group D had no fluoride exposure. The fluoride concentration was determined by using a modified micro-fluoride electrode. The concentration of fluoride changed obviously with the delay of reservation. The fluoride concentrations of finger blood tested within one day from the four groups were different (A > B > C > D). The fluoride concentration in serum was lower compared with that in finger blood. The results showed that serum fluoride was correlated with finger blood fluoride within one day of reservation (r = 0.87). The regression equation was Y = 0.62X + 0.023. It was concluded that the determination of fluoride from finger blood could substitute for serum to reflect the level and degree of fluoride burden of body. PMID- 12561583 TI - [Study on the influencing factor of cultivating mungbean sprouts rich in chromium]. AB - Getting organic chromium by cultivating chromium-enriched mungbean sprouts was studied. Chromium content in mung-bean sprouts was determined and the growth performance was observed under different conditions. The result showed that the optimal condition of cultivating chromium-enriched mungbean sprouts was soaking mungbean seeds in 40 mg/kg chromium solution about 24 hours, then cultivating under 25 degrees C for 5 days. The supernatant of chromium-enriched mungbean sprouts and normal mungbean sprouts were tested by scanning with ultraviolet spectrometry within the range of 200-320 nm wavelength. It is found that a characteristic ultraviolet absorption peak appears at 257 nm, and its optical density is descended with the increasing of chromium in the chromium-enriched mung-bean sprouts. PMID- 12561584 TI - [Irradiation of 235 uranium on the growth, behavior and some biochemical changes of brain in neonatal rats]. AB - The effects of enriched uranium on growth, behavior and biochemical changes of brain in neonatal Wistar rats was studied. Neonatal rats were irradiated with a single injection of 2 microliters enriched uranium (containing 0, 1, 5 and 10 micrograms of 235U) into left lateral ventricle of brain at the first day (< 24 h) after birth. The growth and neurobehavior development of the rats was delay and abnormal. The microautoradiographic tracing technique showed that the radionuclide was mainly accumulated in the nucleus, also appeared in the cytoplasm of cells and between cells. The activity of superoxide dismutase (SOD) and the level of endothelin (ET) of cerebellum, cerebral cortex, hippocampus and diencephalons of brain after exposure to alpha irradiation were examined with radioimmunoassay. The data indicated that SOD and ET was elevated by low dose irradiation and distinctly inhibited by high dose irradiation. It was concluded that alpha irradiation could have damages on the developing brain of neonatal rats characterized by changing the sensibility, fragility and compensation of nerve cells. PMID- 12561585 TI - [Effect of manganese exposure on brain development in postnatal mice]. AB - Effects of manganese (Mn) exposure on brain development in postnatal mice were studied. The results were as follows: (1) The gain of body weight, brain weight and the ratio of brain weight to body weight in the high dose Mn-exposed group was significantly decreased. (2) In the Morris Water Maze Test, the average latency discovering the hidden platform was obviously increased every day during the first 5 days in the high Mn-exposed group, but the latency probing the quadrant in which the hidden platform was located previously was reduced at the 6th day. (3) The protein content of brain tissue was significantly decreased in the high Mn-exposed mice, but the activity of AChE in brain was decreased. (4) The immunoreactivity of glial fibrillary acid protein (GFAP) and the average relative density of GFAP-positive products in hippocampus (area CA3) of both low and high Mn-exposed group, especially the high dose ones, was significantly higher than that of the control group. PMID- 12561586 TI - [Factors of occupational stress on the working ability of petroleum workers]. AB - The relationship between occupational stress and working ability and their influence factors have been studied. 1230 oil workers in 122 work types of oil industry were selected and an occupational stress questionnaire (OSQ) was filled. The working ability index (WAI) in the female was significantly higher than that in the male (P < 0.0001). The working ability of petroleum workers became lower with the increasing of age, the duration of working time and stress levels (P < 0.001). The correlation on the labor load, adverse factors in the working environment and WAI was negative. It was concluded that the occupational stress could decrease the working ability of petroleum workers. PMID- 12561587 TI - [Study on the genotoxicity of dibromoacetic acid in drinking water]. AB - Dibroacetic acid (DBA) is a byproduct of drinking water disinfected by halogenation and it belongs to halogenated acetic acids. It also can be found in ozonated drinking water. Four short-term assays were performed to detect the genotoxic activity of DBA. In the Ames test, DBA showed mutagenic activity in Salmonella typhimurium strain TA100 with or without rat liver-derived metabolic activation system (S9). In UDS test, DBA was found to induce DNA damage in rat primary hepatocyte. Mouse bone marrow micronucleus test indicated that DBA induced the damage of chromosome. In NIH3T3cell micronucleus test, DBA increased the number of micronuclei in cells. Our results suggest that DBA induce DNA damage and has genotoxic activity. PMID- 12561588 TI - [Effect of airborne fungi on interleukin-4 mRNA expression in the peripheral blood mononuclear cells of rats]. AB - The effect of indoor airborne fungi on interleukin-4 (IL-4) mRNA expression in the peripheral blood mononuclear cells (PBMCs) of rats was studied by inoculating via trachea with fungi suspension. A simplified competitive RT-PCR method was used to measure the IL-4 mRNA expression in plasma. The results showed that interleukin-4 expression was much higher in the affected groups than that in the control one. It suggested that IL-4 might play an important role in the sensitization progress of fungi. PMID- 12561589 TI - [Study on cadmium-induced hepatocellular apoptosis in rats]. AB - The effect of cadmium chloride on hepatocellular apoptosis in rats was studied in vivo at the doses of 5, 10 and 20 mumol/kg by using TUNEL (TdT-mediated dUTP Nick End Labelling), DNA ladder electrophoresis and flow cytometry. The results showed that TUNEL-positive cells could be detected in cadmium-treated rat liver cells. DNA fragmentation (ladder pattern) in cadmium-treated liver cells could be displayed by agarose gel electrophoresis. The rates of apoptotic cells in the liver of cadmium-treated rats at these doses were higher than those of control rats significantly. It was suggested that cadmium chloride could induce hepatocellular apoptosis of rats at certain doses in vivo. PMID- 12561590 TI - [Effect of multi-micronutrient on heat adaptation and its probable mechanism]. AB - A multi-micronutrient compound is composed of ascorbic acid, thiamin, riboflavin, vitamin B6, niacin, folic acid, K, Na, Ca, Mg, Cu, Mn, Fe and Zn. In order to explore the effect of the multi-micronutrient on heat adaptation and its probable mechanism, heat-exposed Drosophila melanogaster, mice and rats were observed. The results showed that the multi-micronutrient could significantly increase the survival rate of heat-exposed Drosophila melanogaster and prolong the survival time and swimming time of mice under high temperature. The results indicated that the significant influence of multi-micronutrients on heat adaptation could be the result of enhancing the glucocorticoid receptor activity in hepatic cytosol in the heat-exposed rats, inhibiting liver phospholipase A2 activity in the heat exposed rats, increasing serum SOD activity and decreasing serum MAD content in the heat-exposed rats, as well as increasing level of HSP70 in the heat-exposed Drosophila melanogaster. The results suggested that the effect of enhancing heat adaptation might be related to improving neuroendocrine regulation, enhancing anti-oxidative function against the damage of free radicals as well as increasing HSP70 content. PMID- 12561591 TI - [Effect of Ce3+ on reactive oxygen species in cells]. AB - The effect of Ce3+ on the content of (reactive oxygen species ROS) in hepatocytes and V79 cells was studied with microfluorometry. The results showed that Ce3+ could enhance the concentration of ROS in hepatocytes in high dose and reduce the concentration of ROS in low dose. These results suggested that Ce3+ might decrease the damage of lipid peroxidation and DNA in low dose and might increase the damage of lipid peroxidaion and DNA in high dose. PMID- 12561592 TI - [Effects of zinc deficiency on the distribution of elements in the tissue of pregnant rats and their fetuses]. AB - The influence of zinc(Zn) deficiency on the distribution of elements in tissues of pregnant rats and their fetuses was investigated. Pregnant Wistar rats were divided into zinc deficiency (ZD), pair-fed(PF), zinc supplement (ZS) and control (C) groups. ZD rats were fed for 21 days a zinc deficient (0.7 mg/kg) diet and other rats were fed a similar diet supplemented with zinc (100 mg/kg). On the 21st day of gestation, blood was drawn from heart, laparotomy was performed, and liver, spleen, kidney, placentas, fetuses were removed for analysing on zinc, copper (Cu), iron (Fe), manganese (Mn) and calcium (Ca). The results showed that the element levels in blood, organs and fetuses from ZD rats were significantly lower than those from ZS and C rats. Furthermore, the levels of Cu, Fe, Mn in blood, liver, spleen, kidney of ZD rats were significantly lower than those of PF rats. It suggests that there are some coordination relationships in intestinal absorption and tissue distribution between Zn and other elements, and the decrease of elements in the body does not mainly result from the reduction of dietary intake caused by zinc deficiency. The Fe content in placentas and Fe, Ca content in fetuses of ZD rats showed a significant increase as compared with those of rats in other groups. There may be a competitive mechanism on the transportation of Zn and Fe in the placenta of rats. In brief, the distribution of minerals in maternal tissues induced by zinc deficiency might be different from those in fetuses. PMID- 12561593 TI - [Gene polymorphism at apoB locus and the serum lipids profile in children]. AB - The relations of gene polymorphism at the apolipoprotein B locus and serum lipid profile in children was studied in 308 normal 7-11 year-old children, including 151 boys and 157 girls. Blood samples were collected for all subjects, and then the serum and blood clot were separated. Serum lipids, including TC, TG, LDL-C, HDL-C, apoB, apoA I and Lp(a) were detected. Genome DNA was extracted from blood clot, then apoB-Xba I gene polymorphism were tested by PCR-RFLP method. The results showed that the distribution of apoB-Xba I genotype in 308 children accorded with Hardy-Weiberg inheritance equilibrium law. The frequency of heterozygote(+/-) was 13.3%, allele(+) was 0.067. The frequency of allele(+) was closed to the internal and Japanese reports (0.033 and 0.04), but much less than the Caucasians (0.50). This showed a ethnic and population difference in the inheritance variation. The average LDL-C levels of the heterozygotes(+/-) were 2.17 mmol/L, no difference compared with homozygotes(-/-) (2.21 mmol/L, P > 0.05). There was also no difference for the genotype distribution between the hyperlipidemia group and control group, which may be the results of no enough sample size and the sample selection, and so on. On the other hand, in normal children, serum lipids controlled by many genes, the effect of a single gene might be small. More studies and analysis on the relationship between serum lipids and multiple genes in multisites should be the next step. PMID- 12561594 TI - [Effects of chromium and fish oil on insulin resistance and leptin resistance in obese developing rats]. AB - In order to study the effects of chromium and fish oil on insulin resistance and leptin resistance in obese rats, 50 rats were divided randomly into five groups: basal diet group was fed on normal diet, other four groups were fed on high fat diet. In addition to high fat diet, chromium group fed chromium (3 mg/kg BW), fish oil group fish oil(5 ml/kg BW) and chromium + fish oil group fed chromium(3 mg/kg BW) plus fish oil(5 ml/kg BW). Blood samples were collected in the following five weeks from tail each week to determine blood sugar, insulin and leptin. The results showed that blood sugar, insulin and leptin in fish oil group and chromium group were lower than those in high fat diet control group. Chromium and fish oil improved the insulin resistance and leptin resistance. PMID- 12561595 TI - [Research on the relationship between populations' long-term exposure to fluoride in drinking water and bone fracture in China]. AB - There are contradictory reports on the prevalence of bone fractures associated with long-term fluoride exposure from drinking water. The prevalence of bone fracture in six rural areas of China and the exposure of fluoride in drinking water was investigated. The data including medical history and demographic information, bone fractures, fluoride content in drinking water, physical activity, cigarette smoking, alcohol consumption and dietary intakes were collected. A retrospective epidemiological study by using the same design, method, quality control and the same questionnaire was conducted. A total of 8266 male and female over 50 years of age were divided into 6 groups by the fluoride concentrations in drinking water. The subjects in each group exposed to different levels of fluoride (0.25-0.34, 0.58-0.73, 1.00-1.06, 1.45-2.19, 2.62-3.58 and 4.32-7.97 mg/L) were 1363, 1407, 1370, 1574, 1051 and 1501 respectively. It has been confirmed that drinking water was the only major source of fluoride exposure in the studied populations. The total bone fracture rates were 7.41%, 6.40%, 5.11%, 6.04%, 6.09% and 7.40% in each group. Natural bone fracture rates in each group were 3.01%, 2.21%, 1.31%, 1.65%, 1.43% and 3.66% respectively. The prevalence of bone fracture and water fluoride level appeared a U-shaped relationship. The prevalence of total bone fracture and natural bone fracture in the population with fluoride 1.00-1.06 mg/L in drinking water was the lowest, compared with the groups exposed to fluoride higher than 4.32 mg/L and lower than 0.73 mg/L. The highest prevalence of hip fracture was in the group with higher water fluoride (4.32-7.97 mg/L) exposure. In general, the prevalence of hip fracture was lower and stable up to 1.06 mg/L of fluoride in drinking water, and then it appeared to rise. Based on the data collected in this investigation, it is concluded that the long-term fluoride exposure from drinking water higher than 4.32 mg/L might increase the risk of overall fractures as well as hip fractures. The risk of overall fractures and natural fractures might be lower while the water fluoride level is at 1.00-1.06 mg/L, however, no protective benefits of fluoride for the risk of hip fracture was observed. PMID- 12561596 TI - [A dietary intervention study on the hypertensive high risk population in a northern rural area of Beijing]. AB - To investigate the relationship between optimal living mode, healthy dietary pattern and the incidence of primary hypertension, a dietary intervention study on hte hypertensive high risk population was carried out in Fangshan district of Beijing during 1997-1999. The subjects were divided into two groups: control group and intervention group. A 3-day-dietary-recall combined with weighing method and a healthy diet education program were used in the study. The results indicated that the dietary pattern was not reasonable in the hypertensive high risk population. After two-year-intervention, the dietary pattern of people in the intervention group has changed distinctively. Compared with the results in 1997, the dietary risk factors decreased obviously in the intervention group, but increased in control group, such as high energy, fat, oil and cholesterol which can promote hypertension. Furthermore, in the past two years, the number of over weight + obese people increased by 24.6% in control group, but was no change in the intervention group. The intervention group subjects experienced an average decline of 3.7 and 5.1 mmHg of systolic blood pressure(SBP) and diastolic blood pressure(DBP) respectively, and new cases of hypertension observed in the intervention group were significantly lower than in control group. Thus it can be considered that the dietary intervention may be effective for the high risk population and play a very important role in the prevention of hypertension. PMID- 12561597 TI - [Effect of NaFeEDTA fortified soy sauce on iron deficiency anemia in students]. AB - NaFeEDTA fortified soy sauce was given to 300 students with iron deficiency anemia (IDA) for 3 months. The subjects were divided into three groups: control group, low iron supplemented group (Fe 5 mg/d) and high iron supplemented group (Fe 20 mg/d). Hemoglobin (Hb), serum iron, serum ferrtin, erythrocyte protoporphyrin, total iron binding capacity and serum transferrin were evaluated before and after the study. The results showed that Hb, serum iron, ferrtin and transferrin of the intervention groups had improved and erythrocyte protoporphyrin, total iron binding capacity decreased. There were no differences between the two intervention groups after the trial. The parameters of both intervention groups were much better than those of the control group. The results suggested that NaFeEDTA fortified soy sauce had positive influence on IDA. PMID- 12561598 TI - [Changes in dietary pattern and certain nutrition-related diseases in urban and rural residents of Jiangsu province in 1990s]. AB - A study on the relations of dietary pattern and mortality of some chronic diseases in urban and rural residents of Jiangsu Province in the past decade was made on the basis of the results of food consumption survey and disease surveillance. The results showed that food consumption and dietary pattern changed remarkably during the 1990s. Grain consumption was decreased, but the consumption of animal food was markedly increased. Though the score of desirable dietary pattern(DDP) in urban residents was more than 90, the deducted scores owing to over-consumption of animal food were increased. The mortality form infectious diseases was evidently decreased. While the death rates of some chronic diseases, such as diabetes, hypertension, coronary heart disease, colorectal cancer and breast cancer was increased rapidly in the urban residents. Sufficient attention should be paid to the negative effects of the change of dietary pattern on the health, especially of urban residents. PMID- 12561599 TI - [Comparative analysis on the hearing ability of children in iodine deficiency areas born before and after iodine supplementation]. AB - According to the detection of hearings ability of children in iodine deficiency areas who was born before and after iodine supplement, the effects of iodine supplement on hearing system development was evaluated. METHOD: Hearing was tested by AS-72 type pure zone diagnostic audiometer(made in Denmark). Hearing of 11-14 years old students were tested before iodine supplement in iodine deficiency areas in 1984, the control was students living in non-iodine deficiency areas. Iodine salt (50 mg/kg) were supplied by the end of 1984, the hearings of children born after one year of iodine supplied were tested in 1999. The result showed that the average hearing threshold of students before iodine supplied in iodine deficiency areas was significance higher than of non-iodine deficiency areas. The hearing of children born after one year of iodine salt supplied in deficient areas had no significant difference from that of normal areas. The development of hearing system might be deteriorated by iodine deficiency during pregnant. It was able to meet the need of iodine that pregnant women ate 1:20 thousands iodine salt. PMID- 12561600 TI - [Complementary feeding practice in Chinese rural children]. AB - The information of complementary foods consumed by 14649 children aged from 4 to 24 months in poor rural areas was analyzed. On average, it was at 6 months old for children to start complementary feeding. Approximately one third of children started complementary feeding at 4-6 months old and 16.4% children were fed cereal foods before 4 months old. Only 50% of children aged 12 months and above consumed vegetables/fruits daily and 38% of them consumed animal foods. However, 15.8% children never ate animal foods, and 20% children were fed formula milk daily after weaning time. The impact of family dietary pattern on children's diet increased along with the growth of children. It is obvious that the irrational food pattern would undermine growth of children and such diet behavior would eventually result in malnutrition. Poor complementary feeding practice in some selected rural areas in China played a negative role on children's growth. Hence, nutrition education to the mothers and caretaker should be emphasized so that to improve their awareness of the importance of good nutrition practice and guide them to feed weaning children reasonably. PMID- 12561601 TI - [Determination of arsenic in Chinese herbs by hydride generation atomic absorption spectrometry]. AB - Arsenic in Chinese herbs was determined by Hydride Generation Atomic Absorption Spectrometry (HGAAS). The method was simple and fast, and could be applied to great amount of samples. The detection limit of arsenic was 0.020 mg/kg (dry weight). The recovery and relative standard deviation (RSD) of all samples were 81.5%-125% and 6.2-18.4% respectively. The determined arsenic content of GBW08501 SRM was 82.4%-114.7% of the certified value, and the RSD was 12.6%. PMID- 12561602 TI - [Rapid detection of enterohemorrhagic Escherichia coli in food with multiplex PCR assay]. AB - Rapid detection of enterohemorrhagic Escherichia coli (EHEC) in food was achieved by using multiplex PCR assay. The assay was designed to amplify the 1109, 302 and 228 bp regions of corresponding virulence genes eaeA, hlyAB, slt1 and/or slt2, by using specific primers for each one reaction. In 60 EHEC and other strains, the above three virulence gene sequences were co-amplified from DNA isolated from 12 EHEC O157:H7 strains, 1 O26:H11 strain and 1 O111:H8 strain; eaeA sequence was amplified from DNA isolated from 2 EAEC strains; slt1/2 sequence was amplified from DNA isolated from 1 VTEC strain. No specific amplification product was produced from other 43 non-EHEC strains. For the detection of EHEC in food, results could be obtained within 8 hrs, and the sensitivity of assay was less than or equal to 1.6 cfu/g(ml) of food. The method was applied to 126 food samples, and EHEC was detected in 3 samples. PMID- 12561603 TI - [Studies on the determination of serum thymic factor by capillary zone electrophoresis]. AB - The determination of serum thymic factor (FTS) by capillary zone electrophoresis was described. The optimum ionic concentration and the pH value of buffer system was examined through orthogonal analysis. The optimum conditions found to be 0.05 mol/L sodium tetraborate, 0.01 mol/L phosphate buffer at pH 8.70, working voltage at 12 kV and wavelength at 200 nm. The recoveries for FTS standards added to the bovine serum albumin and serum samples were measured. The values of the relative standard deviation(RSD), recovery and detection limit were 7.62%, 80.28% and 40 ng/ml respectively. The development of this method provide another way for studying the relations of zinc and thymic factor and investigating the effect of zinc on immune function. PMID- 12561604 TI - [Study on the elimination of bacterial endotoxins by ultrafiltration device]. AB - An external-pressed polysulfone hollow fiber UF membrane, was made by phase inversion method. The cutoff molecular weight was 6000. The water permeation rate of this membrane module (psi 90 x 500 mm) is more than 150 L/hr.atm. The operating results show that the elimination of bacterial endotoxins by this membrane has reached the national standard in pharmacopoeia, and this technology is worthy to be extended over to medical industry. In addition, the essence of pyreotogen, the quantification units and measure methods for pyreotogen, and the operating conditions of UF device in the process of eliminating pyreotogen are introduced. Some critical points in operation was discussed. PMID- 12561605 TI - [Analysis on the surveillanca data of newborn death from 1990 to 1999 in Rugao City, Jiangsu province]. AB - A surveillance and data analysis on the cause and death rate of newborn death in Rugao city, Jiangsu Provinces was conducted from 1990 to 1999. There were 1300 cases died in these 10 years. The mortality rate was 0.92% and the proportion of male to female was 1.53:1. More cases (75%) were died in the early stage after birth. The causes of the newborn death were in the order of premature, asphyxia, pneumonia, congenital anomaly and accidental asphyxia. More babies (75.62%) were died in hospitals. The emphasis on newborn health care was pointed out and had discussions. Taking comprehensive measures would be the key point to reduce newborn mortality rate. PMID- 12561606 TI - [Harmful effect of ultraviolet radiation and sunscreening cosmetics]. PMID- 12561607 TI - [Comparing the two neurobehavioral toxicological methods on the toxic effect of methyl mercury]. AB - To compare the two experimental methods for testing neurobehavioral toxicological effects, methyl mercury chloride at the dosage of 0.00, 0.01, 0.05 and 2.00 mg/(kg.d) was force-fed to pregnant Wistar rats during the 6-9th day of conception. The offspring of the 4 groups were randomly selected for Y maze or operant behavior test at the age of 7 or 10 weeks, respectively. The reinforcements of the performance were electric shock for Y maze and food pellets for operant behavioral test. The results showed that no sex-dependent effects were observed both in maze and operant behavior tests. The performances of offspring in 3 exposure groups were worse than that of control group both in Y maze and operant behavior tests(P < 0.05). It was demonstrated that a clear dose effect relationship in maze test on error time(rs = 0.257, P < 0.05), arrival time(rs = 0.216, P < 0.01); in operant behavior test on differential reinforcement of high rate (rs = -0.7273, P < 0.01), but not on differential reinforcement of low rate(rs = -0.2238, P > 0.05). It is concluded that prenatal methyl mercury exposure caused some negative effects on the development of nervous system. But the negative reinforcement by human operation in Y maze test could affect the results. The positive reinforcement and automatic monitoring in operant behavioral test gave more reliable results. PMID- 12561608 TI - [A pilot study on arsenic removal facilities for central supply of drinking water]. AB - In order to promote the application and industrialization of an arsenic removal agent developed in this project, and to improve the technology, it is necessary to have a median sized facility to test the effects, capacity and regeneration of the agent. A facility with the capacity of 1 m3/h for arsenic removal was installed in a bottled water plant, which has been operated since April 1999. The results showed that the arsenic content of effluent water has met the requirement of the national standard for drinking water. The effect of arsenic removal was by 2 mg/g. The annual cost per person was only 1.46 Yuan RMB. The operation and maintenance of the facility was simple and no need to adjust the pH of influents and effluents. The chemical regenerative agent was safe, low cost and effective. Therefore, the facility can be used in rural areas and the water plants while the arsenic in source water was higher than 0.05 mg/L. PMID- 12561609 TI - [Study on the defluoridation of drinking water with synthetic hydroxyapatite]. AB - Synthetic hydroxyapatite was used as a material for the defluoridation of drinking water. The defluoridation capacity, regeneration the capability of defluoridator and the corresponding parameters of defluoridation and regeneration have been studied with batch and column test. The results showed that the defluoridation capacity of synthetic hydroxyapatite on F- was 3.7-4.3 mg/g in batch test, 5.6 mg/g for break through (1.0 mg/L is considered as the break through point) in column test, and 10.7 mg/g for saturation in column test. The efficiency of regeneration was 46%-64% for saturated hydroxyapatite with surface coating method. PMID- 12561610 TI - [Field study on the removal of arsenic from dispersed drinking water]. AB - Shanyin county, an area prevalent of endemic arsenic poisoning in Shanxi Province, was selected as the experimental field to verify the methods studied for removal arsenic from dispersed drinking water. The results showed that the desired goal has been achieved from the field study. The results confirmed that arsenate could be removed from drinking water by using ferric sulfate or aluminum sulfate as coagulants and employing sand filtration. Javelle water(sodium hypochlorite) is an effective oxidant for arsenite. The performance of iron oxide coated sand developed in this study was good for arsenic removal. Further study and application is needed. PMID- 12561611 TI - [Effect of zinc deficiency and zinc replenishment on HOX3.5 gene expression in mouse embryos]. AB - The effect of zinc deficiency and zinc replenishment on HOX3.5 gene expression in mouse embryos was investigated. Sixty Kunming female mice (25-30 g) were divided into three groups: zinc-deficient groups (ZD), zinc-replenished group (ZR) and zinc normal controlled group (ZN). ZD mice fed with ZD diet (Zn 3.0 +/- 0.5 mg/kg), ZN mice fed with ZN diet (Zn 30 mg/kg), and ZR mice fed with ZD diet at first and then with ZN diet from the 7th day of pregnancy. After feeding different diet for 25 days, the mice were made to copulate (female: male = 2:1). At the 12th day of pregnancy, the mice were killed and the embryos were stored at -70 degrees C. The content of mRNA of HOX3.5 in the embryos were determined by in situ hybridization with digxigenin-labeled cDNA probes. The result showed that the expression on area density and L-density (average optical density) of Hox3.5 in ZD and ZR groups were decreased in comparing with those in ZN group (P < 0.05). It was suggested that the decreased HOX3.5 gene expression by zinc deficiency might influence the normal growth of mouse embryos, and this effect might occurr in the early stage of embryo development, which can not be prevented by replenishing zinc at the 7th day of pregnancy. PMID- 12561612 TI - [Inhibition the growth of human leukemia cells by Lycium barbarum polysaccharide]. AB - The effect and the mechanism of Lycium barbarum polysaccharide (LBP-X) on inhibiting the growth of human leukemia HL-60 cells were examined. LBP-X(20, 100, 500, 1000 mg/L) could inhibit the growth of HL-60 cells in dose-dependent manner and decrease the membrane fluidity of the cell. Agarose gel electrophoresis of DNA from the cells treated with LBP-X revealed a "DNA ladder" and positive TUNEL test. The results showed that the apoptosis of HL-60 cells induced by LBP-X maybe its important mechanism on anti-tumorgenesis. PMID- 12561613 TI - [Extraction of the antioxidative substances from Chinese rose and its antioxidative effect on lard]. AB - The antioxidative effect of extracts from Chinese rose was measured with Na2S2O3 I2 titrimetric method by using lard as substrates. It was found that the antioxidative activities of the Chinese rose extracts retrieved by 95% ethanol solution were the highest, and the activities were strengthened with the increase of dosage. Citric acid and amino acid showed synergistic effects in antioxidative activities with extracts of Chinese rose. Ascorbic acid and alpha-tocophrol also showed strong synergistic antioxidative effect with the extracts of Chinese rose. The combined antioxidative activities were higher than the activity of tea polyphenols, and almost as good as that of tertiary butyl P-dihydroxy bengene(TBHQ). PMID- 12561614 TI - [Distributions of body mass index of Chinese adults]. AB - Based on the Third China National Nutrition Survey conducted in 1992, the distribution of body mass index (BMI) in 44,485 adults aged 20-60 y was analyzed, and the cutoff points of BMI for overweight and obesity were also explored. The results showed that the BMI of Chinese adults was in logarithmic normal distribution. The average BMI of this population was 22.1 +/- 3.1. The 50th, 85th and 95th percentiles of BMI were 21.6, 24.9 and 27.4 kg/m2 respectively. The 85th percentile of BMI was much closer to the BMI cutoff points for overweight recommended by WHO, but the 95th percentile was much lower than WHO's cutoff points for obesity. The distribution of BMI based on age, gender and urban/rural areas was different and with statistical significance. It is considered that age and gender should be taken into account while setting up BMI cutoff points for the overweight and obesity of a population, as well as considering the relationships among BMI and morbidity and mortality, body fat composition, and some blood biochemical indexes. PMID- 12561615 TI - [Effect of folic acid and supplemented with vitamin A and vitamin E on depressing teratogenesis induced by cyclophosphamide]. AB - The effect of folic acid and the combined effect of folic acid(FA) + vitamin A + vitamin E on the prevention of teratogenesis induced by cyclophosphamide during pregnancy were observed. The results showed that cyclophosphamide resulted a decreased weight and length of the fetuses, an increased rate of neural tube defects and a retardation of ossification centers. The neural tube defects rates in the groups with no any supplementation, supplemented with folic acid alone and supplemented with FA + vitamin A + vitamin E were 87.27%, 42.93% and 30.08% respectively(P < 0.01). The supplementation of FA or the combined supplementation resulted also in greater fetal weight and length, increased ossification of supraoccipital bone (P < 0.01). The study suggested that the combined effect of FA, vitamin A and vitamin E was superior to FA alone. PMID- 12561616 TI - [Inhibition effect of soybean isoflavones on the oxidative modification of low density lipoprotein]. AB - In order to investigate the effect of soybean isoflavones(SI) on the oxidative modification to low-density lipoprotein(LDL) and to differentiate the effect of SI and alpha-tocopherol, in vitro and in vivo test were conducted. An in vitro model of LDL oxidative modification induced by copper-ion was established by monitoring the production of thiobarbituric acid-reactive substances (TBARS) and conjugated dienes after SI or alpha-tocopherol was added. The in vivo test was conducted by feeding rats with a high fat diet supplemented with SI and measured the sensitivity of LDL oxidative modification mediated by Cu2+ in vitro. The results revealed that when SI was added into the in vitro LDL oxidation system, the content of TBARS or conjugated dienes in the system was much reduced with a dose-effect relationship, whether lipid oxidation being initiated or not by copper-ion at 37 degrees C. In comparison with SI, only a significant inhibiting effect on lipid oxidation while alpha-tocopherol was added before the initiation of oxidation. High fat diet induced a rising of LDL sensitivity of oxidative stress, and adding SI to the high fat diet could counteract the sensitivity of LDL oxidative modification significantly. It is concluded that SI is a valuable natural antioxidant different from alpha-tocopherol in inhibiting LDL oxidative modification both in vitro and inv vivo. PMID- 12561617 TI - [Effect of dietary protein on the function and morphology of diaphragm in rats with experimental emphysema]. AB - The effects of dietary protein on the function and morphology of diaphragm of rats with emphysema were observed in 50 SD rats, 10 rats in each group. The emphysema in groups E(emphysema control), HP(emphysema with high protein diet) and LP(emphysema with low protein diet) was induced by intra-tracheal instillation of elastase (750 U/kg BW). The control rat (C1, C2) were instilled with normal saline. After 6 weeks of experiment, the total lung volume and the average area of alveolus was significantly increased in the group E compared with group C1 (P < 0.05) and the number of alveolus per unit area was also reduced obviously (P < 0.05). The average contractile force of diaphragm at a stimulus rate of 20 Hz expressed as a ratio of the maximal contractile force (F20/Fmax) was increased significantly in group E compared with group C (P < 0.01). The cross-sectional area of slow-twitch fibers increased significantly in group E. After 10 weeks of oral supplementation with 24% casein(HP) or 8% casein (LP) to the rats with emphysema, the contractility of the diaphragm in group LP was lower than that in the control group and the HP group. It was concluded that high protein diet might be beneficial to maintain the function of diaphragm in patients with emphysema. PMID- 12561618 TI - [Research on the dynamic variation and elimination of nitrite content in sauerkraut during pickling]. AB - To investigate the dynamic variation and elimination of nitrite content in pickles and sauerkraut during pickling, green tuber growing in Fuling was selected to make semi-manufactured mustard according to the process technology. Green tuber, cabbage and cauliflower were selected as raw materials to make sauerkraut in family way. The nitrite content in pickles and sauerkraut was determined by spectrophotometry with 1-Amino-2-(a-naphthy lamine) ethoine dihydrochloride. The nitrite content in mustard was 258.8 mg/kg when 6% salt was used and pickling for 8 days; the nitrite content was 235.9 mg/kg when 15% salt was used and pickling for 15 days. There was no nitrite in mustard when pickling for 35 days. The nitrite content in the sauerkraut was 258.7 mg/kg when pickling for 6 days and no nitrite was found after 20 days. The higher nitrite content in pickle and sauerkraut could be eliminated by water. The elimination ratio was 91% for leafy sauerkraut and semi-manufactured cut mustard and only 72% for whole mustard. It was concluded that pickles and sauerkraut that pickled for enough time (for leafy vegetable more than 20 days, for tuber more than 35 days) had no harm for people to eat, The higher nitrite content in pickles and sauerkrauts could be reduced by 91% with water for several times. PMID- 12561619 TI - [Mutagenic activity of organic pollutants extracted from waters in Guanting Reservoir and Yongding River]. AB - Water samples collected from Guanting Reservoir and Yongding River in July 1999 were examined by Ames test to evaluate the mutagenic activity of organic pollutants. The samples have been concentrated by using XAD-2 resin filled columns. In the test with strain TA98, direct mutagenic effects (-S9) were observed in water extracted from Bridge No.8 and Yanchi at the dosage of 0.8 L/plate, indicating the existence of direct frame shift mutagens. The mutagenic effects disappeared after addition of S9, indicating no pro-mutagen in the samples. In the test with strain TA100, direct base pair substitute mutagenic effect was observed only in the sample from Bridge No. 8 at the dosage of 0.4 L/plate. When adding S9, suspicious positive reaction could exist at the dosage of 0.8 L/plate. The results demonstrated that the water in Yongding River has been heavily polluted by mutagens, for which further studies should be carried out to identify the sources and types of these mutagens. PMID- 12561620 TI - [Survey on the microbial pollution of pure water in water dispenser and its disinfection]. AB - The counts of bacteria, total coliforms and fungi of pure water in the water dispensers were detected and the regular pattern of bacterial reproduction in water were studied after the dispensers and water were disinfected. The results showed the bacterial pollution in the water dispensers was serious and the bacteria counts increased with the time of using dispensers lasted. The disinfectant could killed effectively the bacteria in the water sources but it could not control the new bacteria polluted and reproduced while the water dispensers were in use. It was suggested that the bacteria polluted from air should be prevented by taking effective methods and selecting appropriate organic materials for making dispensers. PMID- 12561621 TI - [Mutation of beta 3-adrenergic-receptor gene and the response to dietary intervention in obese children]. AB - In order to study the relationship between mutation of beta 3-adrenergic-receptor gene and the response to dietary intervention in obese children, 47 obese children were randomly separated into two groups. One was dietary intervention group(36 subjects); the other was control group(11 subjects). The subjects were investigated by dietary survey and anthropometry before and after dietary intervention carried out for 3 months. The mutation of beta 3-adrenergic receptor gene resulting from the replacement of tryptophan by arginine at position 64(Trp64Arg) in encoding amino acid residues was detected by using a polymerase chain reaction-restriction fragment length polymorphisms. The results showed that dietary fat was higher (40.7% of energy from fat) in the obese children. After dietary intervention, the increase of weight(kg) and BMI(kg/m2) were significantly lower in the obese children without mutation than in control group(P < 0.05), but the changes of weight and BMI in the obese children with mutations were similar to those of control group. It was concluded that the Trp64Arg mutation of beta 3-adrenergic receptor gene might be one of the reasons why the effect of dietary intervention was poor in some obese children. PMID- 12561622 TI - [Influences of rearing style on the intellectual development of infants]. AB - The factors causing the imbalance on intellectual development of infants in the early period after birth were studied and some better styles of rearing infants were found. The developmental quotient in 68 infants of 3-months-old was tested with Gesell Developmental Schedules and the style for rearing infants was obtained by inquiring their parents. The results showed that: (1) the development of adaptive behavior, language and personal--social behavior was better than that of gross motor and fine motor; (2) giving infants opportunity to move could make the development of adaptive behavior, gross motor and fine motor better. The development of fine motor could be hampered if a pair of mitten was put on hands to avoid scratching infant own face. Out doors activities were more important on the development of adaptive behavior, gross motor, fine motor and personal- social behavior; (3) in addition to the mother, with the cooperation of grandparents or father to look after infants was better. The higher education level of parents always associated with a better mental development of infants; (4) the complex structure with more people in a family make the development of language better. It is suggested that if parents have more knowledge on caring infants, give infants more opportunity to move and communicate with other people, it would be beneficial to the development of infants. PMID- 12561623 TI - [Survey on the fumonisins intake and the urinary Sa/So ratio of people suffered from a high incidence of esophageal cancer]. AB - To explore the correlation between fumonisins and human esophageal cancer (HEC), the estimated fumonisin intake and the urinary sphinganine/sphingosine (Sa/So) ratio were investigated in the population with a high HEC incidence in the suburb of Hebi city, Henan province. Fumonisins were detected from 95 percent of corn samples in the endemic area, and the concentration of total fumonisins (fumonisin B1, B2 and B3) was in the range from 7.2 to 726.8 micrograms/kg. The estimated daily intake of fumonisins was in the range from 0 to 4.84 micrograms/(kg.d) in males, while from 0 to 4.04 micrograms/(kg.d) in females. Compared with the population in a low HEC incidence area, Suixian county, Henan province, the urinary Sa/So ratio of males in the endemic area was statistically significant higher, although no difference was observed in females. The results indicated that fumonisin might be one of the risk factors for the occurrence of HEC. PMID- 12561624 TI - [Synthesis of hapten 3-phenoxybenzyl-2,2-dimetheyl-cyclopropane-1,3 dicarboxylate]. AB - In order to establish the immunoassay for insecticides of pyrethroids, hapten (3 phenoxybenzyl-2,2-dimethyleyclopropane-1,3-dicarboxylate) has been synthesized from chrysanthemumcarboxylic acid via chloridization, esterification and oxidation. The structure of the product was determined by IR, 1HNMR and MS analysis. PMID- 12561625 TI - [Analysis on the dermal irritation caused by imported cosmetics]. AB - Dermal irritation tests on 902 kinds of cosmetics were conducted. The acute dermal irritation/corrosion tests for 175 samples showed no irritation in 65.1% of them, and 34.9% of them were considered as slight irritation. The chronic dermal irritation/corrosion tests for another 727 samples showed no irritation for 32.2% of them, and 67.8% of them were considered as slight irritation, most of which are lipsticks, eye shadows, fluid powder foundations and powder foundations. PMID- 12561626 TI - [Evaluation on the growth and development of children from gangsan primary school in Kundulun district of Baotou City]. AB - In order to evaluate the growth and development of primary school children and the effect of anemia on body height and weihgt, a routine examination on anthropometry and heomoglobin was carried out among the children from Gangsan Primary School in the Kundulun District of Baotou City, including 580 boys and 561 girls for anthropometry and 579 boys and 560 girls for hemoglobin in the Spring of 2001. The results showed that the growth and development of children were higher than the average results of children published in 1992 National Nutrition Survey, and also higher than the results from the Chinese school children physique development in 1995. The average prevalence of anemia was 26.2%, and the girls suffered from anemia were much higher than the boys (30.9% and 21.8% respectively). The highest prevalence of anemia in boys was in the second grades(29.3% and 28.9%), however, the highest prevalence of anemia in girls was in the second grade(43.2%). Compared with normal children, the children suffered from anemia were in lower height, lower body weight and lower body mass index significantly(P < 0.001); The present study indicated that anemia might have adverse effects on the growth and development of children. PMID- 12561627 TI - [Determination of total, soluble and insoluble dietary fiber in foods by enzymatic-gravimetric method]. AB - For studying the contents of dietary fiber in general foods and functional foods, a enzymatic-gravimetric method recommended by AOAC was established in our laboratory. The method for the determination of total, soluble and insoluble dietary fiber in foods and functional foods could be used for many other kind of foods. The relative standard deviations (RSD) of reproducibility between-run and within-run were 2.04%-7.85%, 3.42%-55.23% respectively. The repeatability of the methods was good, and the methods are suitable for many foods. PMID- 12561628 TI - [Advances on the biological effect indices for fine particles (PM2.5) in air]. AB - Fine particles are complex mixture of air pollutants containing not only a great deal of organic compounds but also varied kind of metals, such as B(a)p, Pb, Cd, Cr etc. Most of those compounds are toxic. Some of them are the cause of lung inflammation and asthma, some are genotoxic with a potential of carcinogenesis. Because the fine particles can be inhaled into lung and deposit in lung tissue, which have adverse effects on human health. Epidemiological studies indicated that increased mortality and morbidity, especially for cardiovascular and lung diseases, was associated with the amount of ambient fine particles. Although some scientists thought fine particles damaging the body through oxidant stress, inflammation and genetic materials and so on, the mechanism for them is not known well. Therefore, it is necessary to explore the biological effects of fine particle on human health in the future. PMID- 12561629 TI - [Advances in the study of thermal environmental conditions and health in China]. PMID- 12561630 TI - [Some clinical and radiographic features of tuberculosis in young adults and its early diagnosis]. AB - Clinical and X-ray findings were analyzed in 123 patients aged 18-35 years who suffered from primary tuberculosis. Adenopathy was infrequently encountered and in most patients it appeared as multiple pulmonary forms. Some specific features were revealed. There were seldom limited processes and predominantly exudative and necrotic forms involving mainly the lower lobe. The characteristic features were the frequent involvement of the sixth segment of one or two lungs in the process and the upper-lobar tuberculosis. In 79.7% of the examinees, areas of tuberculous lymphangitis were defined, which preceded the process in some patients or accompanied it or were identified after resorption of parenchymatous changes in other patients. The above specific features of pulmonary processes may be useful for early diagnosis of primary diagnosis in young adults. PMID- 12561631 TI - [Experience with kipferon use in the treatment of patients with pulmonary tuberculosis]. AB - Kipferon that is a combination of recombinant human (2-interferon and a complex of immunoglobulins G, M and A, was used in suppositories as an auxiliary agent in the routine chemotherapy in 36 new cases of pulmonary tuberculosis. A control group included 19 patients identical in sex, age, and the pattern of pulmonary tuberculosis. The clinical, X-ray, and laboratory indices (primarily cellular immunity) were studies before and 1 and 3 months after treatment. The beneficial effect of kipferon was manifested by a more rapid arrest of symptoms of total intoxication eliminated after 2 weeks in 39% of patients in the experimental group and only in 21% in the controls. Normalization of blood parameters occurred following a month in 58.3 and 47% of patients, respectively. Mycobacteria tuberculosis disappeared in the sputum smears following a month of treatment in 62% of those isolating bacteria in the experimental group and only in 37.5% in the controls (P > 0.1; t = 1.6). Positive lung X-ray changes as resolved infiltration, the reduction and closure of caverns were more pronounced in the patients of the experimental group. The most characteristic change in the parameters of cellular immunity during kipferon was a short (as long as 1-1.5 months) decrease in RBT to FGA, which was noted in 47% and 6.7% of patients in the experimental and control groups, respectively (P < 0.01) and which was followed by an increase in the count of CD8+ cytotoxic lymphocytes. This is indicative of the enhancement of these mechanisms of immunity and a reduced need of enhancing or maintaining the activity of proliferative reactions of immunocompetent cells under the conditions of a favourable influence on the course of tuberculous infection. PMID- 12561632 TI - [Interaction of types of drug resistance and immunity in patients with pulmonary tuberculosis]. AB - The authors carried out a comparative clinical and immunological study of different types of drug-resistance (mono-, poly-, MDR of primary and secondary genesis) and immunity in 54 patients with a specific process in the lung. The level of suppression, immunodeficiency, and the incidence of adverse effects were found to be on the increase in the group with the preserved sensitivity to the that with mono-, poly-, and MDR. Impaired immune homeostasis was more pronounced in patients with secondary MDR versus those with primary MDR. PMID- 12561633 TI - [Surgical treatment of diffuse pulmonary emphysema: state-of-the-art]. PMID- 12561634 TI - [Results of surgical treatment in patients with drug-resistant tuberculosis]. AB - Analysis of the results of surgical treatment in 105 patients with pulmonary tuberculosis revealed that 25 patients had postoperative complications. Complications, outcomes, dysfunctions, and life quality were analyzed in late postoperative periods after 4 types of operations: segmentectomy (n = 52), lobectomy (n = 26), pulmonectomy (n = 12), palliative operations (n = 15). The best immediate and long-term results were achieved after segmentectomy in patients with antibiotic-sensitive forms of MBT. PMID- 12561635 TI - [Reconstructive surgical procedures on the bronchi in patients with tuberculosis]. PMID- 12561636 TI - [Cerebral sarcoidosis: clinical aspects and course]. AB - Central nervous system sarcoidosis lesion is rare among patients with extrapulmonary tuberculosis. The lesion was encountered in 3 of the examined 120 patients with this condition. These 3 patients with cerebral sarcoidosis are used as an example to describe the clinical manifestations, diagnosis, and treatment of the disease. Its diagnosis is difficult. When such symptoms as fever, headache, thirst, etc., much alert is required in order to recognize CNS lesion as soon as possible. Its treatment has its own features. PMID- 12561637 TI - [Clinical effects of smoking]. AB - The paper analyzes the data available in the literature on the prevalence of smoking and its impact on different systems of the human body. Basic documents of the World Health Organization on smoking control policy are presented. PMID- 12561638 TI - [Analysis of the efficiency of treatment and causes of death in nosocomial pneumonia in the Republic of Tatarstan]. AB - A comprehensive study was undertaken to examine morbidity due nosocomial pneumonia. An epidemiological survey demonstrated a growth in morbidity and mortality from this disease among the adult population of the Republic of Tatarstan. Pharmacological and epidemiological survey indicated that antibacterial therapy was in conformity with the federal protocols in 43.9% of cases, revealed the optimization of the use of antibacterial agents after introduction of the federal standards. At the same time the low social status of patients dying from pneumonia was accompanied by their appeal for medical aid, which leads to late hospitalization and untimely intensive therapy. PMID- 12561639 TI - [Differential diagnosis of tuberculous spondylitis under the conditions of a specialized sanatorium]. AB - A total of 477 patients admitted to hospital for tuberculous spondylitis were examined. Further studies revealed that the process in the vertebral column was regarded as degenerative and dystrophic changes in 140 patients, nonspecific osteomyelitis in 94, and cancer in 22. The basis of diagnostic errors was the similarity of some clinical and X-ray manifestations and the pathomorphosis of tuberculous spondylitis, older patients, the wide use of antibiotics, which changes the clinical course of spinal tuberculosis. The causes of great difficulties in the differential diagnosis of tuberculous spondylitis lie in the insufficient diagnostic capacities of district tuberculosis dispensaries and in the clinical and X-ray manifestations of spinal tuberculosis which have changed today. PMID- 12561641 TI - [Epidemic significance of trans-ovarian transmission of the pathogens of tuberculosis of different types adapted to the avian body]. PMID- 12561640 TI - [Immunity indicators and the degree of adenosine deaminase activity in patients with pulmonary tuberculosis]. PMID- 12561642 TI - [Markers of alteration of tissue structures as indicators of pulmonary tuberculosis activity]. AB - Some indices that reflect the damaging processes in the cells and tissue structures have been found in 152 patients with pulmonary tuberculosis and in those with inactive lung tissue changes. There is evidence for that increases in the activity of plasma 5'-nucleotidase, ceruloplasmin/transferrin ratio and the appearance of free radical compounds in the plasma are accompaniments of active pulmonary tuberculosis. It is found that the greatest changes in these parameters are, the more acutely the disease runs, the more pronounced an exudative component in the general picture of inflammation is. If there are positive changes in these indices during combined therapy, the activity of 5'-nucleotidase becomes normal just after 5-6 months, and the values of other indices tends to normalize after treatment termination. A combined determination of the ceruloplasmin/transferrin ratio and the plasma levels of free radical compounds allows the activity of a tuberculous process to be established in 70% of cases. PMID- 12561643 TI - [Genetic variants of haptoglobin in patients with nephrotuberculosis]. AB - Clinical, X-ray, ultrasound, and laboratory studies were made in 152 patients with nephrotic tuberculosis. Haptoglobin phenotypes were defined in the examinees. Nephorotic tuberculosis was found to occur in patients with haptoglobin 2-2 most commonly. There was a relationship between the carriage of a genetically determined haptoglobin phenotype and a difference in the clinical manifestations of hephrotic tuberculosis. The patients with haptoglobin 2-2 were seen to have more severe hephrotic tuberculosis. PMID- 12561644 TI - [T cell subsets undergoing apoptosis and anergy in patients with pulmonary tuberculosis]. AB - T-cell apoptosis and anergy as possible causes of impaired antigen specific responses and their subpopulation targets in patients with pulmonary tuberculosis were investigated. A decrease in PPD-stimulated proliferative responses were revealed in 43% of the examinees. The impaired PPD response was shown to be associated with both increased lymphocytic apoptosis and the arrest of cell cycle progression. CD4 and CHD8 T cells underwent apoptosis in PPD-stimulated cultures. Whereas a moderate apoptosis of CD4 cell could occur in PPD-reactive patients, accelerated apoptosis of CD8 cells developed only in PPD-unresponsive group. Both T-cell subpopulations displayed a decreased count of cells in S,G2/M phases of a cell cycle. Similar to apoptosis, the anergy of CD8 T cells was typical of PPD unresponsive patients. Elevated apoptosis and anergy of CD4 and CD8 T cells in vitro were accompanied by a decline in the proportion of T cells and their subpopulations in patients with impaired PPD responses. PMID- 12561645 TI - [The Moscow Regional Extrapulmonary Tuberculosis Sanatorium Krasnaia Roza (Red Rose): 75 years old]. PMID- 12561646 TI - [How to write a good scientific article for a journal]. PMID- 12561647 TI - [Clinico-diagnostic trial in adults allergic to pollen extracts from Cynodon dactylon (L). Pers]. AB - BACKGROUND: There are no antecedents in Cuba of studies developed to prove the efficacy of on allergy test to pollen Cynodon dactylon (L) Pers. OBJECTIVE: To evaluate the allergenic extract pollen Cynodon dactylon (L) Pers elaborated at the reference hospital William Soler City Havana. MATERIAL AND METHODS: We selected 100 patients of 18 to 50 years with a positive clinic history of inhalant allergy and 100 healthy person. All the persons selected were skin prick tested with dilutions of the extract (1:100 and 1:100 P/V) and positive and negative control. We measured the average diameter of the wheal and the perpendicular diameter, then we calculated the average. We also calculated size of the wheal in the duplicated, which constituted the size of the reaction. We estimated the validity of the test calculating the sensibility and specificity. We used the Epitable program to do this. RESULTS: The skin prick tested positive in 81% of the patients and 11% in the healthy person, for sensitivity of 81% the interval of confidence 71.7, 87.9 (CI) and specificity of 89%, CI 80.8, 94.1; the predictive positive value 88.0%, IC 79.2, 93.6 and the predictive negative value 82.4%, IC 73.6, 88.8. CONCLUSIONS: The allergenic extract of pollen Cynodon dactylon (L) Pers is very sharp to test allergy to its pollen produced by the allergology laboratory of Hospital William Soler, and is effective in diagnosing alterations caused by this pollen. PMID- 12561648 TI - [Allergen sensitization and asthma in children from 1 to 3 years of age]. AB - BACKGROUND: Asthma is a multifactorial disease. Children with atopy history could be sensitized with indoor and food allergens from early stages of the life and this predispose to develop asthma. Controversy exists about the factors that increase or decrease the risk of disease, such as breast feeding, early weaning, smoking exposition, immunizations. OBJECTIVE: To determine if these risk factors and inhaled and food allergen sensitization predispose to asthma development. METHODS: A case-control study was performed. Cases (1 to 3 year-old 74 children) with asthma and control (74 healthy age-matched children) were included, r2 = 0.90. Skin-prick test (SPT) was performed by using indoor and food allergens. Test was considered positive if skin reaction was equal or higher than 3 mm. The results were compared by means of the x2 test. A logistic regression analysis was performed for obtaining odds ratio (OR) (95% confidence interval for each factor). RESULTS: Out of the SPT carried out with aero-allergens, 23 (31%) were positive in the two groups. Regarding the food, 27 cases (36.4%) and 15 controls (20.2%) were positive (x2 = 4.7); p < 0.05, OR = 2.2, IC 95% (1.6-3.1). Atopy was positive in 50 (67.5%) cases and 39 (52.7%) of the controls (x2 = 3.4); p < 0.05, OR 1.8 (1.3 to 2.5). The weaning before the first three months of life was positive in 44 (59.4%) of the cases against 15 (20.2%) of the controls (x2 = 23.7) p < 0.05 with a OR 5.7 (4.9 at 6.6). CONCLUSIONS: We found a probability 2.2 times higher for developing asthma when children were sensitive to at least one food. We also reported that 100% of our sensitized cases with food had received breast feeding during the first four months of life, which is similar to results of other authors where the breast feeding during an intermediate time (one to six months) does not protect against allergic disease. Early introduction of solid food (before the three months of age) was the main factor increasing probability for developing asthma. In our study, atopy almost duplicated this probability. PMID- 12561649 TI - [New prospects for the treatment of systemic lupus erythematosus]. AB - The systemic lupus erythematosus is a significant therapeutic challenge: Multiorgan involvement and a variable disease course, characterized by clinical exacerbations and remissions, make difficult to predict the outcome. Few products have been specifically developed for this affection, and most accepted therapies have not been tested in randomized controlled trials of systemic lupus erythematosus. A variety of biological agents under investigation as potential treatments for systemic lupus erythematosus are designed to interfere with specific immunologic responses, hopefully avoiding generalized immunosuppression. Agents, which interfere with T cell-B cell collaboration, such as CTLA-4-Ig and anti-CD154 ligand monoclonal antibodies, may result in long-term therapeutic benefit. Products designed to decrease production of anti-dsDNA antibodies or inhibit complement activation, may prevent immune complex deposition and ameliorate organ-specific manifestations, such as renal disease. More aggressive interventions include gene therapy and stem cell transplantation. PMID- 12561650 TI - [Allergic Rhinitis and its Impact on Asthma (ARIA) in Latin America]. AB - Allergic rhinitis is the commonest chronic respiratory disorder in children and young adults having an important impact for those suffering this condition, as well as for the public health. Allergic rhinitis is frequently associated to other co-morbidities, particularly asthma and conjunctivitis but, also, sinusitis and otitis media. Most of patients suffering rhinitis are cared by GPs and pediatricians and there are evidences that allergic rhinitis is undertreated, particularly the moderate/severe persistent forms. Clinical guidelines have become an important tool providing recommendations for diagnosis and treatment of different medical conditions. They help the process of decision making for GPs and pediatricians, and many of them, contain an update on basic science and epidemiology. In respiratory medicine, guidelines on asthma and rhinitis are available; however, they do not look at the patients globally and focus the disorder on an organ-specific basis without recommendations on co-morbidities. ARIA, Allergic rhinitis and its impact on asthma, has not been developed only to update specialists in allergy/immunology, otorhinolaryngology and neumology on rhinitis and its comorbidities but, also, to provide recommendations for non specialists. A new classification and severity of allergic rhinitis is proposed replacing the classic perennial and seasonal forms for persistent and intermittent, mild to moderate/severe. ARIA is an initiative in collaboration with the World Health Organization and the master document has been endorsed by many national and international scientific societies and organizations. ARIA is an evidence-based document also stressing on pediatric aspects and providing recommendations for low-income countries. PMID- 12561651 TI - Central nervous system involvement in a child with polyarteritis nodosa and severe atopic dermatitis. AB - Polyarteritis nodosa is a diffuse vasculitis of small- and medium-sized muscular arteries that can involve the vessels of one or several organ systems. We report an 11-year-old boy with severe atopic dermatitis further complicated with erythroderma and cerebellar symptoms. Laboratory studies showed negative antinuclear antibodies and antineutrophil cytoplasmic antibodies; increased serum complement, IgG cryoglobulins 96.8 mg/dL (0-80) and serum IgG 2,160 mg/dL (613 1,295), and positive alpha-cardiolipin test. The IgE value was high. Skin prick tests and RAST were positive for dust mites and casein. The viral profile was negative. Imaging studies disclosed tortuousity in the vessels and ischemic infarction in the cerebellum and thalamus. A diagnosis of polyarteritis nodosa with central nervous system involvement was made. The patient improved with immunosuppressive therapy and thalidomide. Currently, the patient is controlled and in a rehabilitation program. Other cases of polyarteritis nodosa associated with atopic dermatitis have not been reported. PMID- 12561653 TI - Physicians, public not overly concerned about medical errors. PMID- 12561652 TI - [Eosinophilic proctocolitis induced by foods. Report of a case]. AB - The food-induced eosinophilic proctocolitis is a major cause of blood-tinged stools and appears in the first two months of life. The infant is generally described as well, but the clinical features and laboratory results are often nonspecific. We present an early infant with fresh blood stools at 50 days. The colonoscopy and biopsy of the rectum and lower sigmoid revealed lymphoid nodular hyperplasia of the submucose and eosinophil infiltration (40 to 50 per high power field) of the lamina propria and intraepithelial. Elimination of the offending protein from the diet, through the use of an extensively hydrolyzed casein-based formula and soy, lead to clinical resolution of bleeding at 48 hours. We made a review of the case. PMID- 12561654 TI - Safety of cancer drug use improves. PMID- 12561655 TI - Report criticizes FDA's oversight of DTC ads. PMID- 12561656 TI - Interest grows in consumer-driven health plans. PMID- 12561657 TI - Improving patient care with collaborative rounds. PMID- 12561658 TI - Projecting future drug expenditures--2003. AB - Drug expenditure projections for 2003 and factors likely to influence drug costs are discussed. The United States continues to face the challenge of increased growth in health expenditures, and drug expenditures are continuing to increase faster than the growth in total health care expenditures. These increases can be largely attributed to an increase in the average age of the U.S. population and technological advancement. On the basis of price inflation and non-price inflationary factors, including increases in volume, shifts in patient and therapeutic intensity, and expected approval of new drugs, a 10-12% increase in drug expenditures in 2003 for the inpatient setting and a 13.5-15.5% increase for ambulatory care settings are forecasted. While few new drugs are expected to greatly influence expenditures in 2003, the continued diffusion of recently approved drugs such as drotrecogin alfa and nesiritide will have a dramatic impact on total drug expenditures and must be carefully considered in the budgeting process. An agent likely to have a significant impact on HIV treatment is enfuvirtide, the first in a new class of antiretrovirals (fusion inhibitors), but its high cost ($10,000-$15,000 per year) may limit patients' access to this medication. An expanded user's guide is provided to assist the reader in appropriate application of this information in the drug budgeting process. Technological, demographic, and market-based changes and changes in public policy will continue to influence pharmaceutical expenditures in the coming year. An understanding of the overall drivers of medication expenditures and vigilance in monitoring pharmaceutical innovation are critical in the effective management of these resources. PMID- 12561659 TI - Documenting pharmacist interventions on an intranet. AB - The process of developing and implementing an intranet Web site for clinical intervention documentation is described. An inpatient pharmacy department initiated an organizationwide effort to improve documentation of interventions by pharmacists at its seven hospitals to achieve real-time capture of meaningful benchmarking data. Standardization of intervention types would allow the health system to contrast and compare medication use, process improvement, and patient care initiatives among its hospitals. After completing a needs assessment and reviewing current methodologies, a computerized tracking tool was developed in house and integrated with the organization's intranet. Representatives from all hospitals agreed on content and functionality requirements for the Web site. The site was completed and activated in February 2002. Before this Web site was established, the most documented intervention types were Renal Adjustment and Clarify Dose, with a daily average of four and three, respectively. After site activation, daily averages for Renal Adjustment remained unchanged, but Clarify Dose is now documented nine times per day. Drug Information and i.v.-to-p.o. intervention types, which previously averaged less than one intervention per day, are now documented an average of four times daily. Approximately 91% of staff pharmacists are using this site. Future plans for this site include enhanced accessibility to the site with wireless personal digital assistants. The design and implementation of an intranet Web site to document pharmacists' interventions doubled the rate of intervention documentation and standardized the intervention types among hospitals in the health system. PMID- 12561660 TI - Immediate free recall of drug names: effects of similarity and availability. AB - The prescribing frequency, subjective familiarity, and two measures of similarity as predictors of error in immediate free recall of drug names were assessed. The study design utilized prospective, computer-based, word memory experiments in which 30 pharmacists and 66 college students were asked to immediately recall 15 lists of three three-syllable drug names. Intralist similarity was systematically varied. The number of words forgotten or incorrectly recalled was then examined as a function of similarity, subjective familiarity, and prescribing frequency. The primary outcome measure was the number of item errors in free recall. Pharmacists made fewer errors than college students. Familiarity reliably enhanced item recall among both pharmacists and college students. Prescribing frequency enhanced recall among both pharmacists and college students except when college students recalled generic names. Orthographic (i.e., spelling) similarity was reliably associated with item recall in both groups. Fewer errors were made when lists were more orthographically similar. Among pharmacists, there was an inverted U-shaped relationship between phonologic (i.e., sound) similarity and item errors, with the fewest errors being made on the most similar lists. Among college students, phonologic similarity was not reliably associated with item errors. Frequently prescribed and subjectively familiar drug names are more accurately recalled than rarely prescribed and unfamiliar names. Orthographically similar lists of drug names are easier to recall than dissimilar lists because similarity provides cues that facilitate the retrieval of degraded short-term memories. The effects of similarity, familiarity, and frequency on short-term memory of drug names vary as a function of task and stimulus characteristics. PMID- 12561661 TI - Cefazolin dosing protocol for patients receiving long-term hemodialysis. PMID- 12561662 TI - Pediatric congenital errors of metabolism: the pharmacist's extended role. PMID- 12561663 TI - Comparison of the political contributions of pharmacists with other health care professionals. PMID- 12561664 TI - Dosage conversions as a potential cause of adverse drug events. PMID- 12561665 TI - Heat-associated increase in transdermal fentanyl absorption. PMID- 12561666 TI - Visual compatibility of doxapram hydrochloride with drugs commonly administered via a Y-site in the intensive care nursery. PMID- 12561667 TI - Corticosteroid-induced adverse reactions in a university teaching hospital. PMID- 12561668 TI - Make pharmacy a medical specialty? PMID- 12561669 TI - Sterility of fat emulsion in plastic syringes. PMID- 12561670 TI - Sterility of fat emulsion in plastic syringes. PMID- 12561671 TI - Immunogenicity and immune response in breast cancer. PMID- 12561672 TI - The assessment of cellular immunity in subjects presenting different immune disorders. PMID- 12561673 TI - Flow cytometric determination of Enterococcus spp. susceptibility to four antimicrobial agents. AB - Two Enterococcus strains (E. faecalis and E. faecium) isolated from 2 patients in an intensive care unit (blood and drain, respectively) were analyzed for susceptibility to 4 antibiotics (penicillin, vancomycin, gentamicin, streptomycin) by agar dilution standard method (MICs), time-kill and flow cytometry. We compared the data from classical methods of antibiotic susceptibility detection, that are compulsory 24 hrs long and flow cytometry results at 5 and 24 hrs cultivation. The results from both classical and flow cytometric analyses were highly cogent and revealed the fact that flow cytometry is very useful in early diagnosis of bacterial resistance to antibiotics. PMID- 12561674 TI - Venous ulcer infection caused by Arcanobacterium haemolyticum. AB - A. haemolyticum has been described as an unusual pathogen causing pharyngotonsillitis and extra pharyngeal infections like ulcerative lesions infection mainly in patients with underlying condition (peripheral vascular disease, diabetes, alcoholism). A case of A. haemolyticum venous leg ulcer infection is reported in a 66 year-old diabetic male patient. Bacteriological diagnosis is based on conventional culture technique. But, due to its coryneform appearance and time growing, it is probably underreported. PMID- 12561675 TI - Involvement of Listeria monocytogenes in the abortive disease. AB - Listeria monocytogenes, an intracellular facultative germ that causes the invasion, sometimes fatal, in susceptible hosts is a food borne pathogen with ubiquitary spread that has generated a public health problem for such risk groups as: pregnant women, foetuses, new borns. 504 women with abortive disease were serologically investigated in 1999 for serotype 1a circulating in Romania. The most affected age group proved to be that in the range of 20-30 yrs: 378 (75%) cases. 107 (21.23%) female patients had the diagnostic titer (> or = 1/320): among these, 38 (7.53%) had miscarriages in the IVth-VIIIth month and 18 (3.57%) gave birth to dead foetuses; during pregnancy, 10 (1.98%) female patients received treatment with Ampicillin and 2 (0.39%) treatment with Erythromycin. In the age group > 31 yrs, the 1/320 titer was noticed in 21 (4.16%) female patients but among these only 4 (0.79%) had a history of miscarriage in the final pregnancy months; they were administered Ampicillin during pregnancy. Although there is no clear-cut evidence, our results point to the conclusion that these female patients were contaminated with Listeria monocytogenes. PMID- 12561676 TI - Characterization of rpo-B mutant of a strain of Escherichia coli to confirm its suitability for routine work related to recombinant DNA technology. AB - Rifampicin resistant spontaneous mutant of a popular laboratory strain of Escherichia coli (DH5 alpha) was isolated and found to resist high level of the drug in growth medium. The growth of the isolate was found to be slower than its wild-type counterpart. Its ability to get transformed into drug-resistant state through transformation by chemical means as tested using plasmid DNA from three different size categories, was found to be at par with the wild type. Other properties, viz., alpha-complementation and ability to express foreign gene remained unaltered. The utility of the rifampicin-resistant phenotype as a potential chromosomal genetic marker was demonstrated in a typical conjugation experiment to establish the ability of the mutant to act as recipient strain for a recombinant, mobilizable plasmid DNA molecule with the advantage of drug mediated, high efficiency selection. Substitution of the wild strain with the mutant for routine experimentations related to recombinant DNA technology was concluded to be appropriate and of advantage. PMID- 12561677 TI - A possible correlation between unscheduled DNA repair and cholesterolemia in Wistar rat, modulated by vitamin E. AB - Cholesterol plays a critical role not only in modulating membrane structure and dynamics but also in its metabolic pathway, to interfere with cell growth and proliferation processes. Having this aim in mind, we have suggested an investigation, by radioisotopic techniques, of the effect of vitamin E (alpha tocopherol, 3 mg/kg b.w. in daily doses, for 7 days) on the unscheduled DNA biosynthesis, induced by Romanian cytostatic Lomustin (Nipalkin or CCNU at the dose of 10 mg/kg b.w in acute administration, for 24 h), both on normally fed animals and on rats having a hypercholesterolemic diet, for 30 days. Considering the scientific data from the literature, according to which there is an inverse correlation between the serum cholesterol level and the risk of developing cancer, we tried to investigate the possible influence of Wistar rat cholesterolemic background on the nuclear unscheduled DNA biosynthesis, essential for the conservation of the cell genome integrity. So, it has been noticed that: 1) the DNA lesions induced by the alkylant and tested by the uptake of 3H Thymidine (200 microCi/100 g b.w.) are reduced after vitamin E treatment, suggesting a protective effect of the antioxidant on the genetic material. 2) on a hypercholesterolemic background the administration of Lomustin produces a decrease of cholesterolemia, suggesting the development of a "facilitating environment" for CCNU action, which appears to confirm the data from the biographical sources. 3) using 3H-Cholesterol (150 microCi/100 g b.w.) to estimate its intracellular liver incorporation suggests a possible displacement of cholesterol from the tissue compartment to the serum one and reverse, event which appears to be correlated with unscheduled DNA biosynthesis. This sustains the idea of the intracellular cholesterol necessity during the nucleic acid biosynthesis as well as in genome aggression. PMID- 12561678 TI - Skin manifestations in parasite infection. AB - BACKGROUND: Intestinal parasites stimulate IgE synthesis by their proteinases. Because of their ability to induce IgE production and mast cell degranulation (either directly, e.g. Ascaris, or via specific IgE) it is possible that some parasites induce allergic manifestations. Some parasites may be more allergenic than others due to their allergenicity. Genetic predisposition, nutritional status, and psychosocial variables of the host and the time and degree of allergen exposure may also play an important role. THE AIM: Of our study was to investigate which parasites are more frequently related to the skin manifestations and if atopic background of the host has a role in these skin allergic hives in intestinal parasites. MATERIAL AND METHOD: 55 patients (13 men, mean age 36 years old) infested with digestive parasites were skin prick tested to inhalant allergens (house dust mites, pollens, animal dander) in order to evaluate the atopic status. All patients had allergic skin manifestations (rash and hives) and angioedema due to their intestinal parasites, which remitted after antiparasitic treatment. RESULTS: Forty (73.3%) patients presented angioedema and 15 patients (26.6%) had chronic rash. Patients who were infected with Ascaris had more severe symptoms: more than 2/3 had angioedema, and only one third had chronic rash. More than 2/3 of the patients were infested with Ascaris lumbricoides: 38 patients (69.09%), and only one third were infested with Giardia lamblia: 18 patients (32.72%). Only one third of the patients (17-30.9%) had positive skin prick tests for at least one inhalant allergen. CONCLUSIONS: The parasite most frequently implicated in skin allergic manifestations in our region is Ascaris lumbricoides. Acute angioedema or chronic rash are the major skin manifestations in parasitic infestation. The atopic trait has no importance in the appearance of allergic manifestations in parasitic infestation. PMID- 12561679 TI - Reviewers deserve better recognition. PMID- 12561680 TI - Homeopathic principles not followed. PMID- 12561681 TI - Not classical homeopathy. PMID- 12561682 TI - A poor test of homeopathy. PMID- 12561683 TI - Studies of homeopathic treatments need to involve both homeopaths and allopaths. PMID- 12561685 TI - Study defies the most basic tenets of homeopathy. PMID- 12561686 TI - An ethicist's commentary on animal rights versus welfare. PMID- 12561687 TI - [Working group of the ACMV on the formation, agreement and enlargement of the door to veterinary medicine]. PMID- 12561688 TI - An evaluation of the relative efficacy of a new formulation of oxytetracycline for the treatment of undifferentiated fever in feedlot calves in western Canada. AB - A field trial was performed under commercial feedlot conditions in western Canada to compare the efficacy of a new formulation of long-acting oxytetracycline (LA 30) to a standard long-acting oxytetracycline formulation (LA 20) and florfenicol (FLOR) for the treatment of undifferentiated fever (UF) in calves that received metaphylactic tilmicosin upon arrival at the feed-lot. Seven hundred and ninety seven recently weaned, auction market derived, crossbred, beef calves suffering from UF were allocated to 1 of 3 experimental groups as follows: LA 30, which received intramuscular long-acting oxytetracycline (300 mg/mL formulation) at the rate of 30 mg/kg body weight (BW) at the time of allocation; LA 20, which received intramuscular long-acting oxytetracycline (200 mg/mL formulation) at the rate of 20 mg/kg BW at the time of allocation; or FLOR, which received intramuscular florfenicol administered at the rate of 20 mg/kg BW at the time of allocation and again 48 hours later. Two hundred and sixty-six animals were allocated to the LA 30 group, 265 animals were allocated to the LA 20 group, and 266 animals were allocated to the FLOR group. The relative efficacy of the LA 30 group, as compared with the LA 20 and FLOR groups, was assessed by comparing relapse, chronicity, wastage, and mortality rates. The overall mortality (RR = 0.50) rate in the LA 30 group was significantly (P < 0.05) lower than in the LA 20 group. However, the overall chronicity (RR = 2.56) and overall wastage (RR = 6.97) rates of the LA 30 group were significantly (P < 0.05) higher than in the LA 20 group. There were no significant (P > or = 0.05) differences in UF relapse rates or cause specific mortality rates between the LA 30 and LA 20 groups. In the economic analysis, there was an advantage of $28.59 CDN per animal in the LA 30 group compared with the LA 20 group. The overall chronicity (RR = 2.25) and overall wastage (RR = 2.80) rates of the LA 30 group were significantly (P < 0.05) higher than the FLOR group. There were no significant (P > or = 0.05) differences in UF relapse rates, overall mortality rates, or cause specific mortality rates between the LA 30 and FLOR groups. In the economic analysis, there was an advantage of $12.90 CDN per animal in the LA 30 group compared with the FLOR group. In summary, the results of this study indicate that it is more cost-effective to use a new formulation of long-acting oxytetracycline (300 mg/mL formulation administered at a rate of 30 mg/kg BW) than a standard long-acting oxytetracycline formulation (200 mg/mL formulation administered at a rate of 20 mg/kg BW) or florfenicol for the treatment of UF in feedlot calves that have previously received metaphylactic tilmicosin upon arrival at the feedlot. PMID- 12561689 TI - The evolution of bovine viral diarrhea: a review. AB - The economic importance of bovine viral diarrhea is increasing with the emergence of seemingly more virulent viruses, as evidenced by outbreaks of hemorrhagic syndrome and severe acute bovine viral diarrhea beginning in the 1980s and 1990s. It appears that evolutionary changes in bovine viral diarrhea virus were responsible for these outbreaks. The genetic properties of the classical bovine viral diarrhea virus that contribute to the basis of current diagnostic tests, vaccines, and our understanding of pathogenic mechanisms are now being reevaluated because of these "new" virus strains. This shift in virulence has confounded both nomenclature and the significance of current bovine viral diarrhea virus categorization. The purpose of this review is to summarize our current understanding of bovine viral diarrhea virus with a chronological review of prevailing scientific tenets and practices as described in clinical and scientific North American veterinary journals and textbooks. The first part of this review describes how we have arrived at our current understanding of the viruses, the diseases, and their nomenclature. The second part of the review deals with current concepts in virology and how these concepts may both explain and predict bovine viral diarrhea virus pathogenesis. By reviewing how knowledge of bovine viral diarrhea has evolved and the theories of how the virus itself is able to evolve, the interpretation of diagnostic tests are more effectively utilized in the control and treatment of bovine viral diarrhea virus associated disease. PMID- 12561691 TI - Veterinarians, the Royal Society of Canada, and the future of veterinary medicine: Part 1. PMID- 12561690 TI - Resurgence of leptospirosis in dogs in Ontario: recent findings. AB - A marked increase in leptospirosis in dogs was observed in 2000, part of an increasing trend observed in previous years in Ontario. The highest frequency of seropositive cases occurred from September to December 2000, with the peak in November. Large breed dogs were particularly affected. Clinical and clinicopathological data for 31 dogs admitted between 1998 and 2000 to the Ontario Veterinary College Veterinary Teaching Hospital were analyzed. Major clinical presenting features were acute onset of anorexia, depression, fever, and vomiting. Ninety percent of dogs, on admission, showed biochemical evidence of injury to several organs, notably combinations in the order of kidney, muscle, pancreas, and liver. Almost all dogs showed increased serum urea and creatinine levels, and the majority had increased total creatine kinase, bilirubin, alkaline phosphatase, and leukocytosis with neutrophilia. One-third were thrombocytopenic. Of dogs with liver-related abnormalities, most had evidence of cholestasis, with or without hepatocellular damage. Based on serologic studies, in the year 2000, the major serovar involved was autumnalis, but bratislava, grippotyphosa, and pomona were also implicated. The microscopic agglutination test often gave a confusing pattern of reactivities to the serovars that were tested. The high reactivity to serovar autumnalis may represent an erroneous or "paradoxical" reaction typical of early leptospiral serology. The year 2000 was the warmest in Ontario in each of the 4 fall months (September-December) of the previous decade, as well as being the third wettest in the fall period in the last decade. The increase in canine leptospirosis, therefore, may, in part, reflect climate change. The number of positive cases declined in 2001 by about one-third of those in 2000, but the number of submissions of sera for diagnosis increased markedly over previous years. Further work is required to isolate and to identify definitively serovars involved in resurgent canine leptospirosis and the common sources for dogs. PMID- 12561692 TI - Diagnosis of a uterine leiomyoma using hysteroscopy and a partial ovariohysterectomy in a mare. AB - A 14-year-old quarter horse mare had a 2-year history of infertility and the presence of a mass in the right uterine horn. Prior to surgery, the mass was determined to be invasive by using ultrasonography and hysteroscopy. A partial ovariohysterectomy was performed. The mass was diagnosed histologically as a uterine leiomyoma. PMID- 12561693 TI - Nonsteroidal anti-inflammatory drug-induced duodenal ulceration and perforation in a mature rottweiler. AB - A mature male rottweiler was evaluated for acute collapse and abdominal pain. The history consisted of concurrent administration of meloxicam and aspirin. On exploratory laparotomy, a large perforated ulcer was discovered in the proximal duodenum, with secondary peritonitis. The pathogenesis of NSAID-induced gastrointestinal ulceration and the supposed safety of COX-2 selective agents are discussed. PMID- 12561694 TI - Capillaria in the bladder of an adult cat. PMID- 12561695 TI - Joint tumors. PMID- 12561696 TI - The argument that never ends. PMID- 12561697 TI - On target. A new generation of drugs offers customized cures. PMID- 12561698 TI - Is there a market for personal medicine? PMID- 12561699 TI - No ordinary life. PMID- 12561700 TI - Portrait: Svante Paabo. The human factor. PMID- 12561701 TI - Pretty horses. Keeping the sea horse from falling victim to its own charisma. PMID- 12561702 TI - [A new stage]. PMID- 12561703 TI - [Risk for congenital malformations in pregnant women exposed to pesticides in the state od Nayarit, Mexico]. AB - OBJECTIVE: To measure the association between the use of pesticides and congenital malformations. MATERIALS AND METHODS: We studied 279 newborn from mothers living in rural area of Nayarit, a state in the northwest region of Mexico. Cases (n = 93) were defined as newborn with central nervous, face, genital, hip, foot or finger congenital malformations. Controls (n = 186) were newborns without any malformation. We considered as exposure any type of contact with any of the agrochemicals used as pesticides. We evaluated other risk factors such as medical drugs, fever, exposure to radiation, obstetric and family factors, as confoundings. RESULTS: We registered 22 genital malformations, 20 from hip, 19 from the central nervous system, 18 from extremities and 14 cleft lip or palate. Exposed mothers had high risk of having a malformed child (OR = 3.5, CI95% 2.05-6.34, p < 0.05). Risk was higher if the mother had occupational exposure to pesticides (OR = 6.33, CI95% 2.95-13.7, p < 0.0001) and in mothers living near areas under pesticides treatment (OR = 3.47, CI95% 1.91-6.33, p < 0.0001). Among obstetric factors, abortion and early delivery (OR = 15.05, CI95% 1.82-124.30, p < 0.01) were significant. CONCLUSIONS: This study shows association between exposure to pesticides and congenital malformation. This is a public health problem in Nayarit state and in other rural areas with similar exposure to pesticides. PMID- 12561704 TI - [Eclampsia in Central America]. AB - OBJECTIVE: To evaluate the incidence, characteristic and maternal outcome of eclampsia in some countries of Central America. METHOD: We reviewed all patients with the diagnosis of eclampsia at the Complejo Hospitalario "AAM" de la Caja de Seguro Social de Panama, Hospital Santo Tomas de Panama, Hospital Bertha Calderon, Managua, Nicaragua and Hospital Pritnero de Mayo, JSSS, San Salvador, EI Salvador from January 1998 through December 1998. Incidence, demographics characteristics, complications, cardinal signs, deaths and cause of death by hospital were analyzed. RESULTS: The total number of deliveries during the study period was 48,645. One hundred two women had eclampsia, for an incidence of 1 in 477 deliveries. Blood pressure was elevated beyond 160/110 mmHg in 39 (38%) patients, and 25% had < 140/90 during the seizures. Proteinuria before seizure was negative in 10 patients. Twenty-six patients had HELLP syndrome. There were 7 maternal deaths in the entire cohort of patients (6.8%), cerebral hemorrhage was cause of death in two women, multiple organ failure in two, disseminated intravascular coagulation in two and sepsis in one. In addition, six deaths had HELLP syndrome. CONCLUSIONS: Eclampsia is directly related to young women, nulliparous, pregnancy without or low prenatal care and gestational age near to term. HELLP syndrome is a high-risk factor to death among patients with eclampsia. PMID- 12561705 TI - [Active management of labor with cesarean index 6.4% and very good materno-fetal outcome]. AB - OBJECTIVE: The objective of this study is to reduce the incidence of cesarean section and to evaluate the impact on fetal-maternal morbidity and mortality by strictly following the guidelines of the National Health Office (Secretaria de Salud) at the Hospital of Gynecology and Obstetrics of Garza Garcia in Nuevo Leon, Mexico. METHOD: A prospective and observational study was made. All the patients who culminated their pregnancy on Saturdays and Sundays between 7:00 and 19:00 hrs, in the period June 2000 to June 2001, were included in this study. The incidence of Cesarean sections and its impact on maternal-fetal morbidity and mortality were observed. RESULTS: A total of 798 patients were included in the study. The total number of newborns was 801. The age average was of 23.4 years (14 years-40 years). Of the 798 patients, 280 (35.1%) presented dystocia at vaginal delivery, 467 patients (58.5%) had an eutocic vaginal delivery and 51 patients (6.4%) went through a Caesarean section. The Apgar scores were: 8 at the first minute and 9 at the fifth minute for 681 neonates (85.01%); and 7/9 for 53 (6.6%). CONCLUSIONS: According to the results obtained in the present study we can conclude that the incidence of Cesarean section can be reduced by strictly following the guidelines of the National Health Office without affecting the maternal-fetal morbidity and mortality. PMID- 12561706 TI - [Clinical competence evaluation in undergraduate gynecology and obstetrics]. AB - OBJECTIVE: Assess the clinical competence in Gynecology and obstetrics to the Internship students of the Faculty of Medicine, UNAM. METHOD: The study design was descriptive, transverse type. We assessed 64 students, which had finished their gynecology field rotation with the objective structured clinical examination. The criteria to consider a competent performance level, was arbitrarily set up in 60%, both for individual problems and for the exam's global result. RESULTS: In 15 stations, the result was a 56.2 global average. The best performances were achieved in the following stations: take the pap smear (74.7), Pregnancy diagnostic (67.9), history of Gynecology and obstetrics (67.1), self examination of breast explanation (62.2) preclampsia (61.7) and cervicovaginitis (60). All the rest got a mark lower than 60. DISCUSSION: The results are lower than the ones obtained in written exams, because these cannot assess clinical skills. It could be observed that a student's performance in a clinical problem does not certainly predict his performance in other, so it seems to be determined more by the specific knowledge and the student's experience related to the case, than by a general problem-solving skill. CONCLUSIONS: The results show the advantages of this instrument to assess clinical skills, that justify its application in the formative process. This work evidences that its necessary to improve the acquisition of basic clinical skills trough systematic instructionals strategies and greater opportunities of learning. PMID- 12561707 TI - [Causes of non-use of contraceptives in the immediate postpartum period]. AB - OBJECTIVE: To determine the causes of not use of contraceptive during immediate postpartum period. METHODOLOGY: Cross-section observational descriptive study. We include women that went for attention of obstetric event, we identified those were in immediate postpartum period, and we selected the women which not started contraceptive use. Data were collected directly with an interview; the causes of not use of contraceptive were classified in three groups. Group I: causes be derived by patient: personal, religious, moral, families, culture, etc. reasons, when they received information, and advice or when they did not attend to educational actions. Group II: causes be derived by the hospital: technique administrative factors, insufficient educational communication activities by service provider. Group III: Medic Indication: presence of risk factors for health women. We found 2,593 women, we identified 1,493 (57.5%) in immediate postpartum period, 478 (32%) not started contraceptive use. In 349 (73%) women the causes were group I, in 91 (19%) group II, and 38 (8%) group III. CONCLUSION: Is necessary more research to know users concerns, ideas and perspectives in relation with methods of contraception, contraceptive counseling, informants, advisers, and with health service institution, to improve educational communication strategies; and to unify medical criterions for not use contraceptive during immediate postpartum period. PMID- 12561708 TI - [Vaginal misoprostol in the prevention of postpartum hemorrhage]. AB - OBJECTIVE: To show the advantages of the use of vaginal misoprostol, a prostaglandine E1 analogue, in the prevention of the post-partum haemorrhage. MATERIAL AND METHODS: This was a prospective, observational, comparative study. The study included 400 patients with high risk of postpartum haemorrhage at our center between January 1999 and may 2001. Patients were divided in two groups. In group I (208 patients) misoprostol was used in a dose of 800 ugr and in group II (192 patients) in whom misoprostol was not used. Both groups were treated initially with our conventional oxytocin protocol. We evaluated the use of additional oxytocin or ergotamine, haemoglobin levels pre and post-partum, the amount of blood loss, and the need for blood transfusion or hysterectomy. RESULTS: The need for additional oxytocin or ergotamine was reduced to less than 10% in group I when compared to group II; the drop in haemoglobin levels and the amount of blood loss were also less in group I (p: 0.03). In this group only one patient needed for blood transfusion and no patient needed hysterectomy. In group II six patients need a blood transfusion and there was the need for two hysterectomies. CONCLUSION: The use of vaginal misoprostol is effective to control the postpartum bleeding, reducing the blood loss after birth in women with high risk of post-partum haemorrhage as well as the need for blood transfusion. It's use has mild side effects and is of low cost. PMID- 12561709 TI - [Menstrual disorders. Amenorrhea, hypomenorrhea, and oligomenorrhea]. AB - After a summary of the physiology of the normal menstrual cycle, the author studies the symptoms of the different menstrual disorders, explaining the physiopathological basis of each. Special attention is given to amenorrhea, hypomenorrhea and oligomenorrhea. This paper is part of a symposium in which the author dealt with this type of disorders. The author insists on the importance of making hormonal dosifications. As far as the treatment is concerned it has to be by means of hormonal products in the correct dosage, and at the appropriate time in the cycle. The author mentions that he has not had experience with radiation treatment. PMID- 12561710 TI - The identification process. PMID- 12561711 TI - Forensic odontology: "where should I begin?". PMID- 12561712 TI - Ancient esthetic dentistry in Mesoamerica. PMID- 12561713 TI - Mass disaster management: the organization of a mass disaster dental identification team. PMID- 12561714 TI - "Be prepared". PMID- 12561715 TI - Bite marks. AB - A simple and practical method for the evaluation of bite mark evidence has been described. There are many other methods, some quite complicated, using overlays, computer analysis and mathematical formulae. This author prefers to adhere to the "KISS principle: "Keep It Simple, Stupid." A jury is generally more willing to accept the positioning of a study model on a one-to-one life-size photograph rather than be led through a complicated analytical procedure. If the juror can hold the model and photograph in his/her hand and see that the teeth and bite mark match (or do not match), data summation can be much more direct. Numerous publications describe more complex matching procedures should circumstances warrant. PMID- 12561716 TI - Forensic odontology in Israel. PMID- 12561718 TI - Patients' rights and responsibilities. PMID- 12561719 TI - Hospital prognosis? PMID- 12561717 TI - Dentistry's great founder, Solyman Brown, and the Jews. PMID- 12561720 TI - Reducing failure rates: system changes lead to better care. PMID- 12561721 TI - Case of the month. Acute frosted angiitis. AB - A 42-year-old woman presented with acute visual loss in the left eye with funduscopic evidence of marked retinal periphlebitis (frosted angiitis) and a macular star. Extensive serologic testing identified no etiology. The purpose of this report is to review the clinical features and systemic associations of acute frosted angiitis, a rare ophthalmologic entity. PMID- 12561722 TI - Early childhood intervention in Arkansas: what role is the gatekeeper? PMID- 12561723 TI - Taking the pain out of student loan repayment. PMID- 12561724 TI - [Extrachromosomal structures containing small nuclear RNP and coilin in the late vitellogenic oocytes of hibernating grass frogs]. AB - We have studied extrachromosomal structures in the germinal vesicle (GV) of the late vitellogenic oocytes of hibernating frogs Rana temporaria. During this period of oogenesis, chromosomes are completely inactivated to be surrounded by a fibrillar karyosphere capsule (Gruzova, Parfenov, 1993). Using immunostaining and in situ nucleic acid hybridization, we have identified three types of extrachromosomal structures: Cajal bodies (CB), nucleoli, and micronucleoli. Immunostaining of GV spreads has revealed that CB and nucleoli contain coilin, a marker protein for CB. The nucleoli were also positively stained with antibodies against Sm-epitope and trimetylguanosine cap of small nuclear (sn) RNP. According to the results of in situ nucleic acid hybridization, the nucleoli contain U6 snRNA. To further investigate a distribution of coilin in GV of the late vitellogenic oocytes of R. temporaria, we injected myc-tagged transcripts of full length human coilin (Wu et al., 1994) into the cytoplasm of oocytes and followed the pathway of the newly translated protein with an antibody specific for the tag. Immunofluorescent staining of spread GV contents demonstrated a specific staining of the nucleoli within 3 h after injection. We suggest that the newly synthesized myc-coilin may be phosphorilated and targeted to the nucleoli. PMID- 12561725 TI - [Morphofunctional changes in hepatocytes during the early postnatal development of rats experimentally infected with the intestinal protozoan pathogen, Cryptosporidium parvum (Coccidia, Sporozoa)]. AB - Morphofunctional changes in hepatocytes of 10-14-day old rats were followed in norm and after experimental infection with different doses of oocysts of Cryptosporidium parvum. The liver index (ratio between the liver and body masses) varied with the intensity of invasion on the background of slowing down up to the total cessation of animal growth rates, and all this obviously pointed to severe pathology. In the infected rats, some cytological indices were shifted compared to the norm: protein amount and the average number of genomes per hepatocyte were seen to increase, the normal ratio between cells with different ploidy levels being violated. The particular correlation analysis was employed to distinguish between the ontogenetic (animal growth related) and pathologic (related to the infection intensity) polyploidization and hypertrophy in hepatocytes. In 10-14 day old rats, the former is affected primarily by the increase in the share of multinuclear hepatocytes, whereas the latter is accomplished by the increase in the number of cells with polyploid nuclei (4c and 4c x 2 cells). In the heavily infected rats, the ontogenetic polyploidy was almost totally suppressed due, presumably, to their growth rate inhibition, the rise in hepatocyte ploidy resulting form the obvious pathological changes in the liver. In the infected rats, the ontogenetic hypertrophy of hepatic parenchymatous cells was not manifested, and the observed protein accumulation in hepatocytes also resulted from the pathological changes in the liver. It is obvious that changes in cell hypertrophy (protein content) may serve as a more susceptible tool that readily perceives the host's stress experienced due to the parasitic infection (cryptosporidiosis), than cell ploidy: the levels of the respective responses of these two parameters differing by 4 times. However, due to the known reversible nature of hypertrophy, it cannot be used for the aims of a long-term prediction about the future mode of liver functioning in the animal that survived cryptosporidiosis. Unlike, such a parameter as frequencies of hepatocytes with different ploidy levels is much more useful in this respect. PMID- 12561726 TI - Genome multiplication of extravillous trophoblast cells in human placenta in the course of differentiation and invasion into endometrium and myometrium. I. Dynamics of polyploidization. AB - Polyploidization of the extravillous trophoblast (EVT) cells at different stages of differentiation and invasion into the uterine wall in human placenta has been studied. An increase in the ploidy level of EVT cells in the course of their differentiation within cell columns (CC) was shown. Stem cells were mainly diploid (86.2%); incidence of polyploid nuclei of highly proliferative cells of the proximal part of CC increased progressively. In the distal part of CC, where EVT cells did not divide mitotically, polyploid cells prevailed, with 58.0 and 3.5% nuclei being 4c and 8c, respectively. The highest percentage of polyploid cells was found in the population of EVT cells attached directly to the surface of the decidualized endometrium: percentage of tetraploid cells turned out to be 74.7% and the share of octaploid nuclei rose up to 4.9%; however, there appeared a few (0.3%) 16c cells. The majority of EVT cells invading the decidualized endometrium were polyploid, the share of octaploid and hexadecaploid cells rose up to 9.7 and 1.4%, respectively. On the other hand, the percentage of diploid cells also increased up to 29.2% as compared to EVT cells attached to decidua (20.0%). The same tendency proved to be even stronger in myometrium: the share of diploid EVT cells increased up to 46.0%, a prominent amount of tetraploid (45.1%) and highly polyploid (8c and 16c) cells retained in the EVT cell population (7.4 and 1.1%, respectively). Immunohistochemical staining of Ki-67 protein (MIB1), which labels cells held in the cell cycle, showed a high incidence of MIB1 positive stem cells (93.7%) and the EVT cells of the proximal part of CC (85.5%) characterized by high mitotic activity. A lower MIB1-positivity (43.2%) was found in the distal part of CC, whereas invasive EVT cells showed no MIB1-labeling. The presence of MIB1-positive nuclei in the distal part of CCs in the absence of mitoses, taken together with data on polyploidization of these cells, indicates their switch to the endoreduplication cycle. As a whole, the data obtained evidence that differentiation of EVT cells of the invasive pathway is accompanied by polyploidization. However, in a population of trophoblast cells capable of most profound invasion (up to myometrium), the proportion of diploid cells rose. These results suggest that the human cytotrophoblast invasion into the uterine wall requires an optimum, not the highest, ploidy level, whereas highly polyploid cells may form a subpopulation at the border between the maternal and fetal parts of placenta. PMID- 12561727 TI - [Flow cytometric analysis of ICRF-193 influence on cell passage through mitosis]. AB - Studying the effect of topoisomerase II (topo II) inhibitors on cell passage through mitosis seems to be important for understanding the role of this enzyme during chromosome condensation and segregation. A flow cytometric assay (Zenin et al., 2001) allowed to determine the mitotic index, and to discriminate between not only cells in G2 and M phases (including metaphase and anaphase cells), but also cells in pseudo-G1 with 4c DNA content. It is shown that topo II catalytic inhibitor ICRF-193 blocks G2-M transition in a lymphoblastoid cell line GM-130. Addition of caffeine to cells abrogated a block of their entering mitosis but not the inhibitor action. Cells entered mitosis, which was proven by the presence of chromosomes in the examined specimen, and, bypassing anaphase, appeared in pseudo G1 with 4c DNA content. We have found that in the presence of ICRF-193 cells, GM 130 and Hep-2 lines, previously blocked by nocodazole when in mitosis and then washed, pass through metaphase, enter anaphase and leave it to pass to pseudo-G1 with the 4c DNA content. Thus, by inhibiting topo II activity ICRF-193 causes abnormal mitotic transition. PMID- 12561728 TI - [Hyperthermia induced signal for apoptosis and pathways of its transduction in the cell]. AB - Hyperthermia (HT) is a physiological agent able to induce apoptosis in normal and tumor cells. The ability of HT to damage different cellular components is underlying the mechanism of apoptogenic activity of HT. The review is aimed to consider the studies representing mainly HT production of genotoxic initial signal and pathways of its transduction to programmed cytotoxic event or apoptosis. In this regard we analyse the membrane effect of HT, HT-induced chromatin and nuclear matrix structure changes, DNA damage, effect of HT on DNA repair mechanism, and the role of heat shock proteins in apoptosis and cancerogenesis. PMID- 12561729 TI - [The route of a bacterium Holospora in the cell of Paramecium (Ciliophora, Protista) from phagosome to the nucleus]. AB - Problems encountered at the initial stages of stable symbiotic system formation are discussed in the review. The most studied models for interaction between pathogenic bacteria and metazoan cells are compared with a similar system including Paramecium (a ciliatte)--Holospora (a bacterium). Literary and our own data on the infection of P. caudatum with specific endocytobionts inhabiting the nuclear apparatus (H. obtusa in the macronucleus), and H. undulata (in the micronucleus) are analysed with respect to the modern understanding of the intracellular vesicle trafficking. PMID- 12561730 TI - [The influence of epidermal growth factor and insulin on proliferation and DNA synthesis in ciliates Tetrahymena pyriformis]. AB - The influence of the epidermal growth factor (EGE) (10(-8) M), insulin (10(-6) M) and EGF (10(-8) M) in combination with insulin (10(-6) M) on proliferation and DNA synthesis in the nuclei of ciliates Tetrahymena pyriformis GL was studied. Insulin and EGF, known to stimulate growth of many types of mammalian cells revealed a mitogen influence on the unicellular eukaryotes. This effect involves stimulation of DNA synthesis, rising synchronization of cell division (upon the influence of EGF), and increase in cell number during the exponential growth. The mitogen effect may be evoked by cell progression in G1-phase under the action of growth factors and, consequently by earlier entry of cells into S-phase of the first cell cycle. Insulin repressed division of cells that entered into the generative cycle. These cells were delayed in late S-phase and G2-phase of the cycle. Part of these cells perished, while other cells could successively overcome the cell block to start their division by the 4th hours of cultivation. A collateral cytotoxic effect of insulin was found, being most prominent in early periods of Tetrahymena cultivation. PMID- 12561731 TI - [Expression of genes coding NGF and BDNF human proteins in embryos of transgenic stocks of Drosophila melanogaster]. AB - Transgenic Drosophila stocks were selected. These stocks contained human ngf and bdnf genes under Drosophila heat-shock promoter. The expression of foreign genes during Drosophila development was demonstrated with Northern and whole mount hybridization. PMID- 12561732 TI - [Protein composition of mesoglea and mesogloeal cells of medusa Aurelia aurita]. AB - Protein composition of mesoglea of the scyphomedusa Aurelia aurita was revealed in SDS-PAGE. Some major bands are visible in mesoglea of a mature medusa: 30, 45 47, 85 kDa, three bands between 100-200 kDa, and several bands with molecular weights > 300 kDa. Polyclonal antisera RA45/47 against protein 45 kDa were raised. RA45/47 react with 45-47 kDa protein in mesogleal sample and protein 120 kDa in mesogleal cells on immunoblot. Immunohistochemical analysis of A. aurita histological sections of young and mature medusae showed antigen localization in mesogleal cell granules and in the apical part of ectodermal cells. In mature medusae, the antigen was localized also in elastic fibers. We can conclude that in A. aurita mesogleal cells, along with ectodermal cells, take part in the formation of extracellular matrix of mesoglea. PMID- 12561733 TI - [Results of estimation of mutation rates for translocation trisomy 21]. AB - Among 1332 cases of trisomy 21 born within 1979-1999 in St. Petersburg, 76(5.7%) were carriers of a translocation between chromosome 21 and other acrocentrics. Among 43 Dq; 21q translocations, 17 were inherited from the mother, and one was inherited from the father, 16 were of sporadic occurrence, and in 9 cases the mode of inheritance was not established. Out of 31 cases displaying Gq;21 translocation, 23 were mutants and 8 of unknown origin. One case of non Robertsonian translocation 21;22 was maternal in origin. It was assumed that the proportion of sporadic cases among translocations of unknown origin is the same as that among translocations of the known origin. However, it is conceivable that the parents of a child with a sporadic anomaly, previously having an uncomplicated reproductive history and healthy children, tend to avoid cytogenetic examination more often than the carriers of translocation. Hence, the reported proportion of de novo cases (-0.6) might be underestimated. The analysis of pregnancy outcomes in mothers of children with Down syndrome, who inherited translocation (n = 12), sporadic translocation (n = 12) and translocation of unknown origin (n = 8), supports this suggestion. Analysis of the data from 8 reports, where the origin of Dq;21 was specified, revealed that in those samples, where the origin was traced in almost all families, the proportion of de novo cases (0.75-0.82) was higher than in samples where an appreciable part of families was not examined (0.46-0.73). Therefore, with the aim of correct determination of mutation rate for Dq;21 translocation, the true proportions in D;21 cases merit evaluation. Meanwhile, using average estimation from all the above mentioned reports (0.67), the mutation rate for translocations Dq;21 in St. Petersburg was calculated to be 1.2 x 10(-5) and 0.8 x 10(-5) in 1980-1989 and 1990-1999, respectively. For Gq;21 translocations/isochromosomes, the corresponding figures were 1.6 x 10(-5) and 1.5 x 10(-5). PMID- 12561734 TI - [Tartrate-resistant acid phosphatase in free-living Amoeba proteus]. AB - Tartrate-resistant acid phosphatase (TRAP) of Amoeba proteus (strain B) was represented by 3 of 6 bands (= electromorphs) revealed after disc-electrophoresis in polyacrylamide gels with the use of 2-naphthyl phosphate as a substrate at pH 4.0. The presence of MgCl2, CaCl2 or ZnCl2 (50 mM) in the incubation mixture used for gel staining stimulated activities of all 3 TRAP electromorphs or of two of them (in the case of ZnCl2). When gels were treated with MgCl2, CaCl2 or ZnCl2 (10 and 100 mM, 30 min) before their staining activity of TRAP electromorphs also increased. But unlike 1 M MgCl2 or 1 M CaCl2, 1 M ZnCl2 partly inactivated two of the three TRAP electromorphs. EDTA and EGTA (5 mM), and H2O2 (10 mM) completely inhibited TRAP electromorphs after gel treatment for 10, 20 and 30 min, resp. Of 5 tested ions (Mg2+, Ca2+, Fe2+, Fe3+ and Zn2+), only the latter reactivated the TRAP electromorphs previously inactivated by EDTA or EGTA treatment. In addition, after EDTA inactivation, TRAP electromorphs were reactivated better than after EGTA. The resistance of TRAP electromorphs to okadaic acid and phosphatase inhibitor cocktail 1 used in different concentrations is indicative of the absence of PP1 and PP2A among these electromorphs. Mg2+, Ca2+ and Zn2+ dependence of TRAP activity, and the resistance of its electromorphs to vanadate and phosphatase inhibitor cocktail 2 prevents these electromorphs from being classified as PTP. It is suggested that the active center of A. proteus TRAP contains zinc ion, which is essential for catalytic activity of the enzyme. Thus, TRAP of these amoebae is metallophosphatase showing phosphomonoesterase activity in acidic medium. This metalloenzyme differs from both mammalian tartrate resistant PAPs and tartrate-resistant metallophosphatase of Rana esculenta. PMID- 12561736 TI - Docs respond to group therapy. PMID- 12561735 TI - [The hormone-sensitive adenylyl cyclase system of the ciliate Dileptus anser]. AB - The hormone-sensitive adenylyl cyclase system (AC system) was found and characterized for unicellular eukaryotes--the ciliatae Dileptus anser. It has been first shown that hormones of higher eukaryotes--biogenic amines (adrenalin, isoproterenol and serotonin) and peptide glucagon--stimulate in dose-dependent manner the activity of adenylyl cyclase (AC) of D. anser. The enzymatic activity was stimulated also by guanine nucleotides--GTP and their non-hydrolysable analogue Gpp[NH]p. Stimulating effects of hormones and guanine nucleotides strongly depend on the level of AC basal activity, which is relatively easy to reach (1430 to 3900 pmol cAMP/min per 1 mg of protein). The sensitivity of D. anser AC system to hormones and guanine nucleotides shows the presence of receptor or receptor-related molecules, capable of interacting with the hormone and activating AC through heterotrimeric G-proteins, in ciliatae. On the base of obtained data, a conclusion is made about the similarity of the structural functional organization of AC systems of D. anser and higher eukaryotes. PMID- 12561737 TI - e-health rises from the ashes. PMID- 12561738 TI - Consolidation in 2002 a serious case of deja vu. PMID- 12561740 TI - Readers perspectives. Wireless networks will be mission-critical I.T. tools for most health care provider organizations in the year 2010. PMID- 12561739 TI - Electronic claims growth remains stubbornly slow. PMID- 12561741 TI - Driving forces in cancer diagnostics. PMID- 12561743 TI - Using a creatinine ratio in urinalysis to improve the reliability of protein and albumin results. PMID- 12561742 TI - Talking with Randy Daniel. Interview by Celia Stevens. PMID- 12561744 TI - Can you keep a secret? Give your lab results a HIPAA privacy checkup. PMID- 12561745 TI - When should a phlebotomist ask for help with a difficult draw? PMID- 12561746 TI - Developing a winning strategy in the neurosciences field. PMID- 12561747 TI - How severe is the nurse shortage? PMID- 12561748 TI - Experts highlight the importance of monitoring cardiology programs. PMID- 12561749 TI - Cap Gemini forecasts health care's top 10 business issues for 2003. PMID- 12561750 TI - HCA wins $8.8 million judgment against Humana for nonpayment. PMID- 12561751 TI - Eating for better vision and healthy eyes. PMID- 12561753 TI - Home cooking. How to get started. PMID- 12561752 TI - Bariatric surgery. An option for long-term weight loss. PMID- 12561754 TI - The healing value of fun. PMID- 12561756 TI - Cooking from the diabetes food pyramid. Milk. PMID- 12561757 TI - Diabetes quiz. How much to you know about carbohydrate? PMID- 12561755 TI - What your doctor is reading. Vitamins. PMID- 12561758 TI - For parents. Talking to your kids about substance abuse. PMID- 12561759 TI - Clinical trials? Should you sign up? PMID- 12561760 TI - Women & diabetes. Top 10 health tips for women over 65. PMID- 12561761 TI - Beyond brushing. Other oral health tools. PMID- 12561762 TI - Limited joint mobility. PMID- 12561763 TI - Supermarket smarts. Lunch meat. PMID- 12561764 TI - Tools of the trade 2002. PMID- 12561765 TI - Handling holiday stress. PMID- 12561766 TI - Cooking from the diabetes food pyramid. Fats, sweets, and alcohol. PMID- 12561767 TI - Handheld computers in diabetes management. PMID- 12561768 TI - Diabetes quiz. How much do you know about handling sick days? PMID- 12561769 TI - Spiritual self-care and the use of prayer. PMID- 12561770 TI - Taking cardiovascular risk factors to heart. PMID- 12561771 TI - A new take on New Orleans cuisine. PMID- 12561772 TI - What your doctor is reading. Hormone replacement therapy. PMID- 12561773 TI - [Complementary Medicine--subtle materialism? Or: the question of consciousness]. PMID- 12561774 TI - Unlocking the mystery of oral cancer: HSDM student researcher finds link in chain of oral carcinogenesis. PMID- 12561775 TI - Creative researcher tests a resin that may reduce patient discomfort. PMID- 12561776 TI - Practice, research and teaching: the HSDM model. PMID- 12561777 TI - Pre-doctoral student examines the mysteries of c-myc. PMID- 12561778 TI - [Cloning and sequence analysis of the phytase phyA gene of Aspergillus niger N25]. AB - The phyA encoding phytase of Aspergillus niger N25 was amplified by the polymerase chain reaction (PCR) with primers designed according to the sequences of the phyA in GenBank. The amplified fragment was cloned and sequenced. The results show that: the coding region is 1506 bp in size, includes a 102 bp intron, and encodes a peptide of 476 amino acid residues, in which there is a signal peptide with 19 amino acids and a mature peptide of 448 amino acids. Comparison of this sequence with the phyA of the natural A. niger NRRL3135 (GenBank Accession: M94550), the most highly secreting-phytase strain, shows that the nucleotide homology is as high as 96.746%, and the amino acid homology comes up to 97.64%. The phyA of A. niger N25 strain in this paper is appropriate to be used to construct the phytase gene-engineering bacteria. PMID- 12561779 TI - [Environmental epidemiology research leads to a decrease of the exposure limit for mercury]. AB - The central nervous system is particularly vulnerable to prenatal exposure to methylmercury. Due to the widespread exposure to methylmercury from fish, several prospective environmental epidemiology studies have been initiated, in which the maternal exposure during the pregnancy is related to the neurobehavioural development of the children. We have studied a Faroese birth cohort prenatally exposed to methylmercury from maternal intake of contaminated pilot whale meat. At seven years of age, clear dose-response relationships were observed for deficits in attention, language, and memory. An increase in blood pressure was also associated with the prenatal exposure level. The exposure limit for mercury has therefore been decreased. PMID- 12561780 TI - Contextual influences on reproductive health service use in Uttar Pradesh, India. AB - This study examines the determinants of the use of four types of reproductive health-care services in Uttar Pradesh, India: contraceptive services, antenatal care, delivery in a medical institution, and services dealing with reproductive tract and sexually transmitted infections. The analysis uses a multilevel modeling strategy to assess the presence of household- and community-level variation in service use. The influence of community-level characteristics and reproductive health-care service attributes on service use is examined. The results highlight strong community-level influences on service use, although the type of community effect varies by service type. The role of some individual and household factors in determining a person's use of services is mediated by the characteristics of the community in which the individual lives. The results demonstrate the need to look beyond individual factors when examining health-care seeking behavior, and illustrate that there is no singular "community" effect on service use. PMID- 12561781 TI - [Diagnostic and therapeutic techniques for arterial diseases]. PMID- 12561782 TI - 2002 year in review. ACHE: a year of progress. PMID- 12561783 TI - [Remarks of the Zurich Discussion Panel on the gene test]. PMID- 12561785 TI - [German Association for Rehabilitation of Disabled Persons: European Specialty Congress "Nursing Need--Challenge for Rehabilitation", October 2001]. PMID- 12561784 TI - [Pharmacogenomics in gynecology. Replies to the Zurich Discussion Panel]. PMID- 12561786 TI - [Change in the perspective of disability policy is a fact--Europe needs an anti discrimination guideline]. PMID- 12561787 TI - Wide-band spectral tuning of heat receptors in the pit organ of the copperhead snake (Crotalinae). AB - Receptors located in the facial pit organ of certain species of snake signal the presence of prey. Infrared radiation is an effective stimulus suggesting that these receptors may be low-threshold temperature receptors. We recorded from the nerve innervating the pit organ of snakes belonging to the family of Crotalinae while stimulating the receptive area with well-defined optical stimuli. The objective was to determine the sensitivity of these receptors to a wide range (0.400-10.6 micro m) of optical stimuli to determine if a temperature-sensitive or photosensitive protein initiated signal transduction. We found that receptors in the pit organ exhibited a unique broad response to a wide range of electromagnetic radiation ranging from the near UV to the infrared. The spectral tuning of these receptors parallels closely the absorption spectra of water and oxyhemoglobin, the predominant chromophore in tissue. Our results support the hypothesis that these are receptors activated by minute temperature changes induced by direct absorption of optical radiation in the thin pit organ membrane. PMID- 12561788 TI - Experience and preliminary results in human genome diversity research. PMID- 12561789 TI - The Human Genome Diversity Project: cogitations of an African native. PMID- 12561790 TI - The HGDP and the politics of bioethics. PMID- 12561791 TI - Ethical opportunities offered by the Human Genome Diversity Project. PMID- 12561792 TI - Reflections on the Human Genome Diversity Project. PMID- 12561793 TI - Global human diversity and ethics. PMID- 12561794 TI - The Human Genome Diversity Project: ethical concerns. PMID- 12561795 TI - Legitimate and illegitimate views of the HGDP. PMID- 12561796 TI - A needed boost for the HGDP. PMID- 12561797 TI - The hazards of studying human genetic variation. PMID- 12561798 TI - In review. AB - In a year that saw the chancellor bestow riches on the NHS beyond its wildest dreams, the rejection of the consultant contract and the furore around foreign medical teams shows all was not easy. Paul Stephenson remembers 2002. PMID- 12561799 TI - Shock to the system. Interview by Paul Smith. PMID- 12561800 TI - Best evidence in anesthetic practice: prevention: dimenhydrinate prevents postoperative nausea and vomiting. PMID- 12561801 TI - Spontaneous rupture of bladder presenting as peritonitis. PMID- 12561802 TI - [Karen Blixen and her physicians]. AB - In March 1941, two months after her wedding, Karen Blixen was diagnosed as having syphilis in the second stage. She was treated initially with mercury and later on in Denmark with salvarsan. Years later she received more treatment with mercury, salvarsan and bismuth, but in fact she was cured already in 1915 and told so by her venerologist Carl Rasch. However, she did not believe him, and several physicians, including well-known specialists in internal medicine and neurology told her many years later that she had to accept the diagnosis tabes dorsalis, i.e., syphilis in the third chronic stage. This paper claims, based on her medical records from several hospitals, that her physicians' attitude resulted in the delay of right treatment for her real disease for many years and led to at least one unwarrented surgical procedure (chordotomy). In 1956 she finally received surgical treatment of her stomach ulcer which for many years had caused her attacks of abdominal pain. The procedure was delayed for ten years because of a lumbar sympathectomy, which removes the pain for some years but not the ulcer itself, nor the bout of vomiting. Many doctors (and biographers) have been puzzled by her life-long bowel symptoms. It was often called tropic dysentery, in spite of the fact that this diagnosis was never confirmed by stool analyses. Instead it is suggested that most likely the Baroness caused the symptoms. She misused strong laxatives during her whole adult life. She did not tell her doctors about this until very late in her life and then it was far too late. Many times barium enemas showed a severe chronic condition with dehaustration and dilatation. The reason for her misuse was the fact that she was afraid of gaining too much weight. She used amphetamine during her life in Denmark after her return in 1931 in order to reduce her appetite, and probably she used Chat in Africa. She also constantly smoked cigarettes which in combination with minimal food intake facilitated the development of her stomach ulcer. It is concluded that Karen Blixen would have had a much better life, if communication between her and her physicians had been better. She should have told them and they should have been better to listen to that which was unsaid. PMID- 12561803 TI - America's other drug problem: how the drug industry distorts medicine and politics. PMID- 12561804 TI - [The first decades of Danish lung cancer surgery]. AB - The paper describes the first decades in Danish lung cancer surgery. The pioneers were Erik Husfeldt, Head of the Department of Thoracic Surgery at Rigshospitalet and Tage Kjaer, Head of the municipal Department of Thoracic Surgery, Oresundshopitalet Copenhagen. Up-to-date operative treatment depended on early diagnosis which required education of the general public and cooperation with other medical specialities, especially tuberculosis out-patient clinics. The results in a geographical region demonstrated that surgery had an effect on 5 year survival, but the problem og lung cancer deaths could not be solved by surgery alone, due to the increasing number of new cases. The second generation of thoracic surgeons were educated by the pioneers, but they were also fully trained in general surgery. New departments were established in local hospitals, whose catchment areas were to small. Therefore, for many years the surgical activity of these departments was not confined to thoracic surgery. The establishment, activities and results of one of these new departments are described. The question remains: When can we tell a lung cancer patient that he has been cured by the operation? The answer according to a cautious estimate is: After fourteen years. PMID- 12561805 TI - Musculoskeletal oncology. PMID- 12561806 TI - Trocar site recurrence in laparoscopic surgery for colorectal cancer. PMID- 12561807 TI - Effect of preoperative radiochemotherapy on lymph node retrieval after resection of rectal cancer. PMID- 12561808 TI - Anal sphincter injury after vaginal delivery in primiparous females. PMID- 12561809 TI - Combined pelvic floor rehabilitation. PMID- 12561810 TI - Radiofrequency coagulation: an alternative treatment in early grade bleeding hemorrhoids. PMID- 12561811 TI - Breast brachytherapy. PMID- 12561812 TI - Unleashing the long-distance PCR for detection of the intron 22 inversion of the factor VIII gene in severe haemophilia A. PMID- 12561813 TI - Latent coeliac disease, hyperhomocysteinemia and pulmonary thromboembolism: a close link? PMID- 12561814 TI - A response to: laboratory testing for thrombophilia in pediatric patients. PMID- 12561815 TI - Obituary: Michael John Rand (1927-2002). PMID- 12561816 TI - [Some knowledge to the art of medicine: a short view over two alternative methods in Iceland]. AB - Homeopathy was taken up by a vicar in Northern Iceland around 1850 and was very well received right from the beginning. Soon other lay-men and folk-healers took up homeopathy, either partly or entirely, and their practice expanded quickly. Hydropathy was brought to the country at about the same time as homeopathy, but it was brought by a physician, Jon Hjatalin, who became Director General of Public Health in Iceland in 1855. He introduced the method to other physicians by his teachings and to the public in his writings. By his untiring preachings about the benefits of water he most likely got the public to think and act about hygiene sooner than they would have done otherwise. Regarding homeopathy, Dr. Hjaltalin, tried repeatedly to point out that those who wanted to practice it, had to have some qualifications but both the population and the authorities saw that as unnecessary and the Icelandic parliament (Altinget) sided strongly with the homeopaths, even to the point of taking a stand against the physicians. The parliament tried to legalise homeopathy and in 1911 it passed a law that made it legal for anybody to practice homeopathy. No licence or proof of qualification was demanded. The result was that almost every folk-healer took up homeopathy, to a lesser or greater degree, with or without much knowledge of the method. A comparison of the number of patients who visited a homeopath and a local doctor in the same area show that as soon as a doctor became easily accessible, and medicine became better and safer, patients preferred the doctor to the homeopath. The numbers also show that the homeopath always kept a number of patients, indicating that there may always be a market for some alternative to doctors. PMID- 12561817 TI - Declaration of Interests for WHO experts. PMID- 12561818 TI - Volume of procedures and outcome of treatment. Danish study found no association between hospital and surgeon volume. PMID- 12561819 TI - Electronic tagging of people with dementia. Tagging should be reserved for babies, convicted criminals, and animals. PMID- 12561820 TI - Melanin, nutrition, and the lion's mane. PMID- 12561821 TI - Electronic tagging of people with dementia. Technologies may be enabling. PMID- 12561822 TI - Questions of methodology. PMID- 12561823 TI - Squamous cell carcinomas of the head and neck. Precancerous lesions in oral cavity of Indian schoolchildren may hint at epidemic. PMID- 12561824 TI - On teething symptoms. PMID- 12561825 TI - Cylindrical channels from concave helices. PMID- 12561826 TI - Quality of impact factors of general medical journals. PMID- 12561827 TI - Efficacy should drive atypical antipsychotic treatment. PMID- 12561828 TI - Screening for aortic aneurysm. National screening programme is long overdue. PMID- 12561829 TI - Haemophilus influenzae type b epiglottitis. Article gives timely lesson. PMID- 12561830 TI - EU reimporting drugs meant for Africa is only part of story. PMID- 12561831 TI - [Mrs. Popp - and all the others: the first patients admitted to the Kommunehospital in Copenhagen on the opening day September 19, 1863]. AB - The gates of the Kommunehospital in Copenhagen opened for the first time on September 19th, 1863. It is recorded that on this day 89 patients were admitted to the hospital. These patients have been subdivided according to occupation, their own or that of the family, as it was recorded on admission. The occupational groups are: journeymen, workers, seamen, servants, self-supporting and paupers. Most of the patients are men, reflecting the fact that on the opening day only male patients were moved from Almindelig Hospital that the new Kommunehospital was to replace. Most of the patients came from the working class, a few belonged to the middle-class. That was the general trend at Danish hospitals in the 1860s. The case records provide information about hospital life and about social conditions in 1863, as regards work and family life. According to an article in the 125 year's Anniversary publication of the hospital, the first patient admitted was a widow Mrs. Popp, who suffered from eczema. This study has shown that probably the first patient was a mate by the name of Frits Nygaard. He suffered from smallpox and was admitted already on the 9th of August to the isolation-house, which was taken into use before the official opening of the hospital. PMID- 12561832 TI - Stem cells mixing may form a human-mouse hybrid: mice with human cells would be likely. PMID- 12561833 TI - Drug makers battle a U.S. plan to curb rewards for doctors. PMID- 12561834 TI - Claim of human cloning provokes harsh criticism: many fear backlash on stem cell research. PMID- 12561835 TI - To study disease, Britain plans a genetic census. PMID- 12561836 TI - All clones are not the same. PMID- 12561837 TI - [How, why, when and for how long time did the Danish veterinarians have the right to dispense drugs?]. AB - Danish veterinarians are still allowed to prescribe and distribute drugs sold by Danish pharmacies to be used for treatment of animals under their care in agreement with the Danish rules and EU directives (4). Since the first student graduated from Danish Veterinary School in 1775, Danish veterinarians have also, in agreement with their education, had the right to dispense drugs produced in their own dispensary. However, due to the development of the methods for drug production, the introduction of registered products from medical companies, and the justified demands for documentation of drug efficacy and safety was abolished in 1988, and no exemptions were granted after January 1st 1990 (1). The controlled use of drugs with documented efficacy and safety has improved the clinical effect of animal treatment, minimized side effects in treated animals and unwanted effects on the environment and the consumers of animal products (22). PMID- 12561838 TI - Sterilization offer to addicts reopens ethics issue. PMID- 12561839 TI - Artificial heart implant leads to suit over consent process: recipient's widow says she and her husband were misinformed and misled on risks, benefits. PMID- 12561840 TI - The grim mission of a Swiss group: visitors' suicides: travelers from all over seek assistance from Dignitas, raising government alarm. Marie Hascoet's final hours. PMID- 12561841 TI - [A Danish nurse serving in Rome and Paris during the post-war years after World War I]. AB - In letters to her aunt a Danish nurse, sister in law to the Danish medical historian professor dr.med Evd. Gotfredsen, describes her life, professionally and as a tourist, in Rome and Paris. In 1920 she lived in Rome at the Dinesen Pension, established around the change of the century by a Danish lady. The pension developed into the favorite resort for many Scandinavian Visitors to Rome, some of them famous. During her stay in Paris 1920-26 she had close relations to the nursing bureau established under the auspices of the Danish Council of Nurses (D.S.R). For one and a half years she was the private nurse of the famous ethnologist Roland Bonaparte, grandson of Napoleon's brother Lucien, during his terminal bladder cancer. PMID- 12561842 TI - Materials samples face rigors of space. PMID- 12561843 TI - Bioreactor grows beating heart tissue. PMID- 12561844 TI - [Spaceflight experiments for the study of gravimorphogenesis and hydrotropism in plants]. PMID- 12561845 TI - [Application of nitrifying and denitrifying processes to waste management of aquatic life support in space]. PMID- 12561846 TI - [Toward the promotion of researches utilizing microgravity]. AB - Although many opportunities to make experiments under microgravity conditions have been given in the recent decade, it does not seem that a dramatic or decisive result has been obtained by use of microgravity. Promising some experiments in the space station are now near at hand, but it may be necessary to reconsider what the microgravity experiment is or how the microgravity field should be effectively utilized. PMID- 12561847 TI - [Video image analysis of respiratory and cardiac activity of tadpoles dedicated to space experiment]. PMID- 12561848 TI - [Effects of the spaceflight on morphogenesis and gene expression in the growing rats]. PMID- 12561849 TI - [Changes in the mechanical and chemical properties of rice coleoptile cell walls under microgravity conditions]. PMID- 12561850 TI - [Role of light as a gravity-substituting factor in growth regulation of plants II -azuki bean epicotyls]. PMID- 12561851 TI - Mutation induction by heavy ion irradiation of gpt delta transgenic mice. AB - Using the new transgenic mice produced by mating gpt delta with p53 knockout, mutation induction by heavy-ion irradiation and the effect of p53 background on such induction were studied. After the whole body irradiation with 10 Gy of 135 MeV/u carbon-ion beam, the genomic DNA was isolated from the different organs and the lambda DNA was rescued as a lambda phage. Mutations in the transgene on the lambda DNA were determined by the spi(-) selection (deletion assay). The spi(-) mutation was induced by the above irradiation, but enhancement of the mutant frequency by the knockout of p53 gene was found not in the phages recovered from liver but in those from kidney. We are now making an effort to determine the nature of spi(-) mutation to confirm such p53 effect. PMID- 12561852 TI - [Space radiation dosimetry by a combination of CR-39 and TLD]. PMID- 12561853 TI - [Effects of a simulated microgravity condition and externally applied phytohormones on cell elongation and expansion in higher plants]. PMID- 12561854 TI - [Effects of microgravity conditions simulated by a 3-dimensional clinostat on osmotic concentrations and mechanical properties of cell walls in relation to plant growth and development]. PMID- 12561855 TI - [Mechanism of automorphic curvature in rice coleoptiles under simulated microgravity conditions]. PMID- 12561856 TI - [Effects of anxiolytics on hemodynamic responses to acute microgravity environment]. PMID- 12561857 TI - [Effects for pregnancy of mice in parabolic flight]. PMID- 12561858 TI - [Activities, which modulate mutability of cells, in human serum after parabolic flight (Report No. 2)]. PMID- 12561859 TI - [The gene expressions and morphogenesis of Xenopus kidney A6 cells cultured in simulated microgravity and hypergravity]. PMID- 12561860 TI - [Gravitational response of Xenopus A6 cells]. PMID- 12561861 TI - [Factors affecting emesis induced by parabolic flight in anurans]. PMID- 12561862 TI - Theoretical analysis of mixed irradiation. AB - We revised the models for mixed irradiation by Zaider and Rossi and by Suzuki, substituting second-order repair function for a first-order one in reduction and interaction factors of the models. The reduction factor, which reduces the contribution of the square of a dose to cell killing in the models, and the interaction factor, which also reduces the contribution of the interaction of two or more doses of different types of radiation, were formulated by using the second-order repair function. These newly modified models for mixed irradiation can express or predict cell survival more accurately than the older ones, especially when irradiation is prolonged at low dose rates. PMID- 12561863 TI - [Effect of hypergravity on X-ray-induced cell cycle arrest in mouse m5S cells]. PMID- 12561864 TI - [Enhanced mutation of HPRT locus by ionizing radiation in combination with high gravity]. PMID- 12561865 TI - [Effect of hyper-gravity on radiation induced mutation induction]. PMID- 12561866 TI - [Directional asymmetry in otolithic nystagmus elicitation]. PMID- 12561867 TI - [Effects of body tilt on the torsional eye movements of goldfish during linear acceleration]. PMID- 12561868 TI - [Studies on the balance sense nervous systems in upside-down swimming catfish]. PMID- 12561869 TI - [The gravity-regulated formation of peg and auxin transport in cucumber seedlings]. PMID- 12561870 TI - Direct observation of boundary fluctuation generated by Joule's heat in isotachophoretic migration. PMID- 12561871 TI - Free-flow isotachophoresis under micro-gravity. AB - When free flow electrophoresis (FFE) is applied to the separation and purification process under microgravity, higher separability is expected than that under usual 1 G environment, because sedimentation of samples and heat convection can be neglected. Although FFE under micro-gravity is a very important technique for efficient separation of macromolecules because of the above merits, FFE unit (FFEU) should be more sophisticated to give stable operation, high separability, high through-put and high separation efficiency. In this study, fundamental research to achieve FFE with such high performance was done by using a commercial FFEU. The research contains (1) the quantitative study on the separation mode of FFE and separation efficiency, (2) the feasibility analysis of free-flow bidirectional isotachophoresis (FFBITP) which enables separation of cations anions simultaneously by isotachophoresis (ITP) and zone electrophoresis (ZE), (3) the optimization of hardware factors such as thickness of the separation chamber which may cause deterioration of separation due to heat convection, and (4) the analysis of hardware factors depending on the long-time operation. The ultimate goal of this study is to obtain fundamental data for the development of FFE for the international space station. In this paper, the experimental results obtained in a reduced-gravity aircraft was mainly. PMID- 12561872 TI - [Clinostat rotation culture modulates gene expression of osteoclastogenesis regulating factors via a cyclic-AMP dependent mechanism]. PMID- 12561873 TI - [Effects of clinorotation on production of PGE2 and expression of COX-1 and COX-2 mRNA in MC3T3-E1 mouse osteoblasts]. PMID- 12561874 TI - [Effect of simulated microgravity conditions on auxin polar transport of transgenic Arabidopsis]. PMID- 12561875 TI - [Thigmotactic behavior of bottom dwelling tadpoles and their behavior under microgravity]. PMID- 12561876 TI - [100th anniversary of the death of Rudolf Virchow (1821-1902). Model of a politically responsible physician]. PMID- 12561877 TI - Role of recombinent interleukin-12 as an adjuvant on vaccine-induced immunity in murine Schistosoma mansoni infection. AB - In the present work, the role of recombinent Interleukin-12 (rIL-12) as an adjuvant to soluble worm antigen preparation (SWAP) in protection against primary murine S. mansoni infection, using certain immunization protocol, was studied. A highly significant statistical increase in resistance (P<0.001) was observed between the group immunized with SWAP and rIL-12 (group I) and those immunized with SWAP only (group II), when each was compared to the infection control group (group III). Moreover, resistance to challenge infection was higher in group I (73.6%) than in group II (66.1%). In comparison to group III, histopathological examination of liver sections of groups I and II showed marked reduction in granuloma sizes, with more reduction in group I to the extent that some ova were seen without cellular reaction around them. Liver necrosis and fibrosis were detected only in the infection control group. In contrast to group III, sections in the small intestine did not show any granulomatous reaction in groups I and II. rIL-12, when administered with SWAP could inhibit mesangial cells proliferation in the kidney glomeruli of mice in group I. However, minimal mesangial cells proliferation was observed in the kidney sections from mice in group II, when compared to the prominent proliferation seen in group III. rIL-12 has a prominent role when administered as an adjuvant to SWAP, against primary murine S. mansoni infection and for preventing granulomatous reaction, decreasing worm burden and increasing resistance to infection. PMID- 12561878 TI - IL-10, IFN-gamma and TNF-alpha in acute and chronic human fascioliasis. AB - The three cytokines, IL-10 (Th2 product), IFN-gamma (Th1 product) and TNF-alpha work in concert. The present work was undertaken to study the level of these cytokines during the acute and chronic stages of human fascioliasis in an attempt to assess the involvement of Th1 and Th2 cells in regulation of the immune response in this disease. Sera of twenty six Fasciola patients were obtained and divided into two groups: twelve with acute and fourteen with chronic fascioliasis, sera of a control group were studied simultaneously. During the acute stage, a significant increase was observed in IL-10, IFN-gamma as well as TNF-alpha and specific antibody level. In the chronic phase, a significant increase of IL-10 level was observed. IFN-gamma showed a lower level as compared to the acute stage but TNF-alpha was still high. Accordingly, in fascioliasis in the early phase of infection B cells, macrophages, Th1 and Th2 cells were all activated. They cooperate in overcoming the parasite and work to the benefit of the host. With time and after maturation of the evading worms, Th2 action predominates. IL-10 (Th2 cytokine) which is antagonistic to IFN-gamma (Th1 product) and consequently to TNF-alpha limits the immunopathology that may be caused by the latter. PMID- 12561879 TI - On Atractis species: a nematode from the caecum of dabb-lizard, Uromystax aegyptius (Anderson, 1896) in Saudi Arabia. AB - Six dabb-lizard, Uromystax aegyptius were caught from Riyadh and Jeddah areas, and examined for natural helminth parasitic infection. They were all parasites free except for innumerable small nematodes which were identified Atractis species. The parasites were collected from the caeum and lower gut. The whole results were discussed. PMID- 12561880 TI - DNA analysis of hepatocytes: a novel method for estimating the value of liver tissue vaccine (LTV). AB - Because of the socio-economic condition of African countries, the traditional types of vaccines should be encouraged. LTV in a preliminary study, proved to be of value in reducing morbidity and mortality in Schistosoma mansoni infection. In the present study and because of the vaccine contains liver tissue homogonate, the present authors analyed the DNA content of the hepatocytes of the vaccinated and control groups of animals to know any effects of the vaccine on the hepatocytes ploidy. The DNA content of vaccinated animals was higher. The results were discussed. It was concluded that for testing S. mansoni vaccine, DNA analysis of heptocytes for detecting proliferative S-phase is an important as a positive immune reaction. PMID- 12561881 TI - Fascioliasis an increasing zoonotic disease in Egypt. AB - Fascioliasis is now imposing itself as a zoonotic disease in sheep and cattle raising countries. In Egypt, human fascioliasis is increasing. During the years 1994 to 1997 the overall slaughtered animals in Egyptian abattoirs was 2,003,200 sheep and goats, 2,624,239 cattle and 3,536,744 buffaloes. The overall rates of fascioliasis were 2.02% for sheep and goats, 3.54% for cattle and 1.58% for buffaloes. Macroscopic examination of sheep liver showed up to 100 flukes per liver inside a largely dilated thick walled bile ducts. Cattle liver showed up to 275 flukes per liver inside thickened dilated and calcareous bile ducts with offensive yellowish brown bile. Buffaloes liver showed up to 330 flukes per liver. Microscopic examination showed mainly thickened wall, hyperplasia and marked fibrosis. The discussion focused on the zoonotic importance of fascioliasis in Egypt, diagnosis, treatment and prevention. No doubt, adequate control of fascioliasis is more or less a problem, since it requires the control of snail intermediate host(s) and control of infection in all affected animals and man. PMID- 12561882 TI - Sensitive and specific assay for detecting circulating antigens in individuals infected with Schistosoma mansoni. AB - Monoclonal antibody 128C3/3/21 in an antigen-capture ELISA was used to detect circulating antigen in individuals infected with Schistosoma mansoni. This antibody recognizes a carbohydrate epitope expressed on the major group of acidic egg glycoproteins and on glycoproteins and glycolipids in all other stages of parasite development. The overall sensitivity of the assay was 78%, with a sensitivity of 100% for patients excreting >100 egg/g feces (EGF) and 72% for those excreting <100 EGF. By increasing the degree of antibody biotinylation, the authors have now achieved sensitivities of 92.4% overall and 82% for those excreting <100 EGF. A direct increase in the mean level of circulating antigen was found with increasing egg counts. The difference between those excreting >100 EGF (53 individuals) and those excreting <100 EGF (39 cases) was statistically significant (P<0.01). None of the control sera (23 uninfected individuals and 16 patients infected with other parasites) had circulating antigen levels >80 ng/ml. Thus, the test specificity was >99%. The test accuracy was 94.7%, the positive predictive value 100%, and the negative predictive value 84.8%. PMID- 12561883 TI - Studies on prenatal infections in children with unknown cause of mental retardation and examination of their mothers. AB - Specific IgG antibodies against Toxoplasma, rubella, cytomegalovirus and herpes simplex (TORCH agents) were investigated in 32 infants and children with unexplained mental retardation (MR) and their mothers. 16 mentally normal infants and children of the same age group and their mothers were chosen as controls. Specific Toxoplasma IgG antibodies were 43.75% for MR cases and 37.5% for their mothers. Six (18.75%) mothers gave history of eating raw meat and canned food, while 8 (25%) had pet cats. 56.25% of the MR cases, and 81.25% of their mothers were positive to specific rubella IgG antibodies. 75% of MR cases and 81.25% of their mothers were positive for CMV specific IgG antibodies. Also, 18.75% of the MR cases and 25% of their mothers showed positivity to herpes simplex specific IgG antibodies. 93.75% showed positivity to one or more of the mentioned pathogenic agents. PMID- 12561884 TI - Natural Lieshmania infection in sand cats captured in Riyadh district, Saudi Arabia. AB - Felis margarita or sand cat is a nocturnal carnivore occurs in sandy areas of desert in many African and Arabin countries. Ten sand cats were captured alive in Riyadh district. Indirect haemagglutioation reactions against leishmaniasis were detected in four of them at dilutions 1/64 (1), 1/512 (1) and 1/1024 (2). On the other hand, Leishmania amastigotes were detected in the splenic smears of four (40%), two of which had amastigotes in the liver smears as well (20%). Unfortunately, culture for typing was not done. The role of sand cats as zoonotic host of leishmaniasis was discussed. PMID- 12561885 TI - Rodenticidal activity of two natural coumarines from ether extract of Citrus bergamia. AB - The rodenticidal activity, expressed by LD50 values of ether and petroleum ether extracts of Citrus bergamina pulp against two rat species, Rattus norvegicus (Norway rat) and Rattus rattus (roof rat) was evaluated. Based on LD50 values, ether extract was nearly 1.6 and 1.3 times more toxic to Norway rat and roof rat than ethanol respectively, while pet. ether extract was non toxic to the two rat species. From ether extract, two natural coumarines, Bergapten and Coumurrayin were isolated and tried against the two rat species as anticoagulant rodenticides. Norway rats were more susceptible to both compounds than the roof ones. A direct parallel relationship was found between the toxicity and changes in the blood picture. PMID- 12561886 TI - A spotlight survey of sandflies of the genus Phlebotomus in Jeddah, Saudi Arabia. AB - Leishmaniasis in the Old World is vectored by phlebotomine sandflies of the genus Phlebotomus. In a survey of sandflies in Jeddah area carried out by sticky traps and CDC light traps, three species were identified. These were P. papatasii, P. bergeroti and P. alexandri. The diagnostic morphology was illustrated. The results were discussed. PMID- 12561887 TI - The efficacy of the volatile oils of three plants for controlling Lucilia sericata. AB - Myiasis is one of the most important parasitic diseases, which affects the human welfare. The volatile oils of three plants were tested against adult Lucilia sericata. The LC50 was 80 ppm by Anethum graveolens, 180 ppm by Conyza dioscoridis and 130 ppm by Mentha microphylla. The slope functions were 1.9, 1.79 and 2.2 respectively. The whole results were discussed. PMID- 12561888 TI - Diagnosis of gastrointestinal nematodes infecting sheep in Qalubia Governorate by infective thrid stage larvae. AB - A total of 300 faecal samples obtained from living sheep in Qalubia Governorate, 128 (42.66%) harboured eggs of nematodes. The gastrointestinal nematodes had been differentiated through the infective third stage larvae. Details of the general and differential categories of each were given. The recovered species were Trichostrongylus axei, T. colubriformis, Ostertagia circumcincta, Haemonchus contortus, Bunostomum trigonocephalum and Oesophagostomum venulosum. PMID- 12561889 TI - Outdoor resting and nectar feeding behaviour of some mosquito species in two villages in Egypt. AB - Mosquito resting on different host plants were collected from two localities in Egypt, Kafr Tohoria village, Qualubiya governorate and El Abtal village, Ismailia governorate. Collected mosquitoes were identified, classified to their feeding stage and tested for carbohydrate feeding. Results indicated that Cx. pipiens represented 96% and above of collected species in the two localities. The available plants and crops showed different attraction levels for resting female mosquitoes. Most of the collected females were empty. Mosquitoes may be attracted to certain plants due to the presence of some phagostimulant chemicals rather than the presence of the sugars. PMID- 12561890 TI - Cryptosporidium infection among sheep in Qalubia Governorate, Egypt. AB - The present study was carried to explore the incidence of Cryptosporidium infection among lambs in Qalubia governorate as well as to study the ability of three staining techniques developed for rapid identification of Cryptosporidium oocysts in faecal samples of 120 neonatal lambs less than one month old. The stains used in this investigation were modified Zeihl-Neelsen stain (MZN), Safranin-methylene blue (SMB) and Giemsa stain (GS) to detect the recovered oocysts with their incidence variation. The diagnostic value was raised by direct smear and concentration floatation technique. The infection rates with Cryptosporidium oocysts among the investigated faecal samples from neonatal sheep were 82, 58, 36, out of 120 (68.3, 48.3 and 30%) by modified Zeihl-Neelsen stain, Safranin-methylene blue and Giemsa stain respectively. The lambs less than one week had higher infection rate. The relationship between age susceptibility and infection was discussed. The use of modified Zeihl-Neelsen stain was currently recommended procedure with the concentration floatation technique for the accurate diagnosis and rapid identification of Cryptosporidium oocysts. PMID- 12561891 TI - Specific anti-Toxoplasma antibodies in relation to infection and reinfection using different infective stages. AB - The possibility of rat re-infection with different Toxoplasma gondii stages was studied in this work. Two groups of rats were infected with either intra peritoneal injection of 1000 trophozoites of virulent Toxoplasma strain or 1000 oocysts through oral routes. IgA, IgM and IgG were detected 15, 30, 60 and 120 days post infections. IgA and IgM appeared as early as 15 days post infection but declined afterwards. IgA antibody response was more prominent with oocyst infection and IgM was more prominent with tachyzoite infection with insignificant statistical difference. On the other hand, IgG started to increase 60 days post infection, then increased gradually till the end of the experiment (120 days). The primary orally infected rats were either re-infected orally with oocysts or intra-peritoneal trophozoites and re-examined after 15 and 120 days for the same immunological parameters. Those rats which primary infected with intra-peritoneal trophozoites were re-examined after oral infection with oocysts or injection of trophozoites intra-peritoneal. The level of IgA and IgM in one rat of group A and 2 rats of group B were significantly increased and this occurred in presence of anti-Toxoplasma IgG from the primary infection. In this study, the rats were lab bred so, the factors affect the immune system could be under control, also strain variability was excluded as the infection was only by one strain, and as the IgG level was high, these rats were presumed to be immune from the primary infection. So, the rats which became positive after challenge most probably re-infected. PMID- 12561892 TI - Studies on house dust mites in Great Cairo, Egypt. AB - Nowadays, house dust mites have showed themselves as one of the health problem worldwide. They have to be considered in the differential diagnosis of allergy particularly in children. House dust mites were collected from eight different areas in Great Cairo. Nine species of mites were recovered from indoors. In order of abundance, they were Dermatophagoides pteronyssinus, D. farinae, Tyrophagous putrescentiae, Acarus siro, Cheyletus malaccensis, Blomia kulagini, Acheles graciles, Ornithonyssus bacoti and Lepidoglyphus destructor. All these mites were collected mainly from Bolak Al Dakrour (28.8%) and Al Wayly (27.6%). The least number (1.8%) and species (only three) were collected from Madent Al Salam. Pet and stray dogs and cats, domestic birds and commensal rodents were more or less reported in the majority of the examined areas. Also, patients with history of allergy of unknown cause, were reported in ten out of 80 houses examined (12.5%). The role played by house dust mites in causing allergic manifestations was important environmental disease agents. PMID- 12561893 TI - Parasitological aspects of Capillaria philippinensis recovered from Egyptian patients. AB - Seven new cases of intestinal capillariasis were detected in Kasr El-Aini University Hospitals, Cairo, during the period from July 1997 to June 1998. A parasitological study concerning the morphological features of Capillaria philippinensis recovered from the stool of Egyptian cases was carried out. Eggs were separated, cultured and used to infect Oreochromis nilotica niloticus, a famous popularly consumed type of fresh water fish in Egypt in a trial for identification of the fish intermediate host for this parasite, but infection had not developed in the intestine of this species of fish. PMID- 12561894 TI - Oxygen free radical and nitric oxide production in single or combined human schistosomiasis and fascioliasis. AB - The levels of superoxide (O(-)2) and nitric oxide (NO) production by monocytes have been measured in 15 patients with S. mansoni, 15 patients with Fasciola and six patients with combined infection as well as in control group (15). The levels of both radicals were significantly higher in all patient groups than in the control group, indicating that these radicals may have a role in the immunity against such infections. Patients with chronic fascioliasis showed lower level of O(-)2 and NO than those with schistosomiasis. This may be due to the lodging of the mature Fasciola spp. away from the immune system and subsequently decreased amount of antigens reaching the circulation. In combined infection, the levels of these products were at the highest value, due to increased antigenic stimulation and cross reactivity between the two parasites which may have lead to augmented immune response. PMID- 12561895 TI - Diagnosis of Blastocystis hominis by different staining techniques. AB - One hundred and fifty stool samples were collected from diarrheic patients of different ages, and examined for Blastocystis hominis by direct smears and concentrated by Sheather's sugar flotation. Staining was done by: Giemsa, two modifications of trichrome stain, modified Ziehl-Neelsen, safranin-methylene blue and two-auramine stains. Out of the 150 cases nine were positive for blastocystosis. The best stains were safranin-methylene blue and modified Ziehl Neelsen stains. They had the advantage of staining cysts and amoeboid forms besides being rapid and easy to perform. The modified trichrome stains identified 8 ie, less specific and were time consuming. The auramine dyes stained the cyst, both the wall and internal body fluoresced brightly. Giemsa stain was not an efficient stain. Scanning and transmission electron microscopy (SEM, TEM) were performed to study the fine ultrastructure. PMID- 12561896 TI - Oral immunization of mice against Schistosoma mansoni using drinking water from trays containing Biomphalaria alexandrina infected with Schistosoma mansoni. AB - Water collected from trays containing Biomphalaria alexandrina infected with Schistosoma mansoni at the time of cercariae shedding (SmISW) and trays containing clean, non-infected, B. alexandrina (NISW) and underground water (UW), were filtered used as a drinking water for 3 groups of albino mice males. After two months, blood samples were collected from the 3 groups and serum was tested for anti-cercarial IgG, then mice were infected with 150 S. mansoni cercariae. Eight weeks after infection, mice were perfused and adult S. mansoni worms were counted. Anti-cercarial IgG was positive in 23 (82.1%) out of the 28 samples collected from mice drinking SmISW and only in 2 (9.5%) out of the 21 samples collected from mice drinking NISW, while all samples collected from mice drinking UW were negative for anti-cercarial IgG (X2=45.897; P<0.001). Worm load was significantly lower in the group of mice drinking SmISW than mice drinking NISW (P=0.032) and mice drinking UW (P=0.02). In mice drinking SmISW, adult worm count showed significant negative correlation with anti-cercarial IgG concentration (Kendall's taub =-0.325 and P=0.018). The results indicate that antigens present in drinking water stimulate a level of immunity against schistosomiasis, (inhabitants of endemic areas) resulting in a lower intensity and severity of infection. Also, it may reduce the specificity of serological tests used for diagnosis of Schistosoma infection, based on antibody determination. PMID- 12561897 TI - Spinal cord schistosomiasis and neurologic complications. AB - Affection of the spinal cord with schistosome eggs, although rare but do occur, causing many neurologic complications. The resulting mass may give rise to controversy in diagnosis. This can be more or less easily judged by MRI. However, radical surgical excision is not always recommended. Biopsied materials for histopathological studies should be done first before any surgical treatment. PMID- 12561898 TI - Studies on the molluscicidal actvity of Calendula micrantha officinalis (Compositae) on fascioliasis transmitting snails. AB - The molluscicidal activity of ethanolic extracts of Calendula m. officinalis (flowers and leaves) was used as plant molluscicides against Lymnaea cailliaudi. The results indicated that flowers extract possess a molluscicidal activity more than leaves extract and the LC50 was 35 and 52.17 ppm respectively. The mortality rate of exposed snails was increased by prolongation of the exposure time. The molluscicidal effect resulted in enhancing energy utilization and nutrient consumption since glucose, lipids, proteins and triglycerids were greatly reduced. The stomach and digestive gland of the treated L. cailliaudi snails were greatly altered. PMID- 12561899 TI - Resistance of Musca domestica (L.) (Diptera: Muscidae) from Gamasa City to some insecticides. AB - Susceptibility tests for house fly Musca domestica collected from Gamasa City and laboratory bred one were carried out. The data suggested that the levels of resistance in Gamasa population against malathion, diazinon, diamethoate (organophosphorus compounds) and permethrin (pyrethroid one) were developed while deltamethrin and cypermethrin were still effective. The highest homogenity response was for malathion followed by dimethoate, deltamethrin and cypermethrin. On the other hand, the highest homogenity response in laboratory population was that of cypermethrin and deltamethrin followed by malathion, dimethoate, permethrin and then diazinon. The EST activity was higher in laboratory population (three heavy bands), than Gamasa one (two faint bands), while the activity of LDH and G-6PDH were higher in Gamasa population than in laboratory one. The ADH activity was the same in both populations. This means that EST was not the main detoxifying target in Musca domestica. PMID- 12561900 TI - Assessment of splenic functions in patients with hepato-splenic schistosomiasis using non-invasive techniques. AB - This study was carried out on forty cases, classified into 3 groups; group I: 10 healthy controls subjects, group II: 20 patients with hepatosplenic schistosomiasis and group III: 10 bilharzial patients who underwent total splenectomy. All cases were subjected to clinical examination, abdominal ultrasonography, rectal snips and laboratory investigations which included: stool and urine analysis, complete blood picture, IHAT for bilharziasis, liver function tests, viral markers, estimaton of T-lymphocyte subpopulations (CD(+)3, CD(+)4, & CD(+)8) by flow Cytometry, silver stained blood films to detect argyrophilic inclusions and 99mTc sulphur colloid splenic scan which was applied to group II only. The present results revealed varying degrees of hypersplenism (anaemia, leukopenia & thrombocytopenia) in GII. Seventy percent of this group was positive for HbsAg, HCV or both in association with schistosomiasis. Abnormal red blood cells (acanthocytes, target cells, pitted cells & normoblasts) and inclusion bodies (Howell Jolly bodies, argyrophilic inclusions & pappenhiemer bodies) were detected with different values in GIII. CD(+)4 cells were moderately reduced in GII while they were markedly decreased in GIII. CD(+)8 cells were elevated in GII and returned nearly to the normal values in GIII with decrease in number of total T-lymphocytes. Most patients of GII showed marked squestration of 99mTc labelled R.B.Cs. in the spleen with reversed hepatic/splenic ratio (normally hepatic/splenic ratio is over two). IHAT showed positivity in 90% of patients in GII while it was 50% in GIII. Although total splenectomy improved the haematological pattern and the cytopenias, which are prominent features in hepatosplenic schistosomiasis, yet the immunological profile was still altered. So, it is recommended to perform segmental splenectomy with retention of a normal mass of functioning residual spleen to preserve more immunological function and to protect against life-threatening occurrence of post-splenectomy sepsis. PMID- 12561901 TI - Plasma levels of nitric oxide in association with severe Plasmodium falciparum in Yemen. AB - One hundred and five patients with Plasmodium falciparum were included, forty three with cerebral malaria and sixty-two without cerebral manifestations. The main clinical presentations in cerebral malaria patients were fever (76.4%), pallor (72%), splenomegaly (60.5%), deep coma (39.5%), jaundice (18.6%), pulmonary oedema (13.9%), subconjunctival haemorrhage (13.9%), severe anemia (Hb<5mg/l) (53.5%), hypoglycemia (glucose<40mg/dl) (67.4%) and haemoglobinuria (6.9%) while in non cerebral malaria patients the clinical presentations were fever (83.8%), pallor (67.7%), splenomegaly (66%), jaundice (9.7%), severe anemia (Hb<5gm/dl) (51.6%) and hypoglycemia (glucose<40mg/dl) (3.2%). Nine patients from cerebral malaria group died after admission. Serum level of nitric oxide (nitrite plus nitrate) were assayed for all patients, serum level of nitric oxide were highly significant in patients with cerebral malaria than those without (34.6 +/- 2.3n. mol/ml VS 12.9 +/- 1.3n. mol/ml; P<0.01). In cerebral malaria, nitric oxide levels were highly elevated in patients with deeper coma than those with lighter coma (48.2 +/- 3.1n. mol/ml VS 24.4 +/- 1.3n. mol/ml; P<0.001) and also higher among patients with longer duration of coma (>72 hours) than among patients with shorter duration of coma (<72 hours) (54.5 +/- 2.8 n. mol/ml V.S. 23.6 +/- 3.1n. mol/ml; P<0.001). Also, nitric oxide levels were correlated with clinical outcome, fatal cases (9 patients) having significantly higher nitric oxide levels than survivors (56.2 +/- 3.1 n. mol/ml VS 32.5 +/- 1.3 n. mol/ml; P<0.001). Thus, higher levels of nitric oxide are associated with indices of disease severity and may predict outcome in-patients with cerebral malaria. These data are consistent with the hypothesis that nitric oxide is involved in the pathogenesis of cerebral malaria. PMID- 12561902 TI - Value of the quantitative buffy coat capillary tube test (QBC) in the microscopic diagnosis of bancroftian filariasis. AB - The diagnostic performance of the commerical quantitative buffy coat capillary tube technique was compared with the standard diagnosis of filariasis by microscopical examination of Giemsa stained thick blood films. The comparison was conducted among 83 subjects. The QBC test was highly sensitive than the thick blood film in detection of microfilaraemic patients. The percentage of positive cases among 35 subjects with fever and enlarged lymph nodes in the axilla or in the groin was 48.6% versus 42.8% by thick blood film. The sensitivity was 94.4% versus 83.3% in 18 subjects with lymphatic varices or hydrocele. The sensitivity of the QBC capillary tubes was poor (20%) versus 6.6% among 30 filarial patients with different degrees of lymphatic edema. It is concluded that the QBC capillary test is very simple, rapid and convenient as a diagnostic method for microfilaraemic patients but not suggested to be used neither in the quantitation of the parasite numbers or in species identification. PMID- 12561903 TI - Epidemiology and immunodiagnosis of schistosomiasis haematobium in low endemic area in Egypt. AB - A survey was performed in Behbeet village in Giza governorate including 370 individuals (172 males and 198 females) representing 10% of the house holds. Clinical, stool, urine and serological tests accompanied by a questionnaire were applied to all participants to find out the prevalence, intensity of infection of S. haematobium, underlying sociodemographic factors, morbidity indicators and the awareness and treatment status among the infected population. It was revealed that the overall prevalence of S. haematobium based on the detection of eggs in urine was 18.1% while the prevalence of antibodies to S. haematobium species specific microsomal antigen was 57.6% detected by enzyme-linked immuno-transfer blot (EITB). The highest age specific prevalence and intensity of infection were detected among school children in the early teenage. Males were at a higher risk of contracting infection than females with a sex ratio of 2.5:1. Occupational and recreational water contact were significantly more frequent among the egg positives than the negative ones. Present history of haematuria and microhaematuria detected by reagent strips had the strongest association with S. haematobium infection followed by leucocyturia and dysuria. Microhaematuria had the strongest negative predictive value (85.7%) in discrimination between egg positive and negative groups while its positive predictive value was the highest (92.9%) when seropositives and negatives were discriminated. Less than half of the infected population were aware of having the disease (43.3% and 41.8% among the egg positives and seropositives, respectively) and lower percentages reported receiving previous treatment for schistosomiasis. No significant differences were detected between groups (P>0.05). Culturally appropriate and effective health education of the population, and training of the staff of rural health units to improve diagnostic and outreach skills are recommended. PMID- 12561904 TI - Evaluation of FAST-ELISA for the diagnosis of experimental trichniosis. AB - In order to compare between FAST-ELISA and ELISA for the diagnosis of experimental trichinosis and study the kinetics of antibody and eosinophilic responses, six New Zeland rabbits were infected orally by Trichinella spiralis larvae. Blood was collected every other day for the first 2 weeks, then weekly for eleven weeks post infection. T. spiralis crude larval antigen was prepared for coating of ELISA plates and FAST-ELISA beads. Blood was examined for eosinophilic count and for serum antibody level by ELISA and FAST-ELISA techniques. The burden of infection was assessed by counting encysted larvae in muscle samples of the infected rabbits. By FAST-ELISA antibodies were detected seven days post infection (P.I.), while with ELISA technique antibodies were detected after 10 days. Both tests detected maximum antibody levels on the 4th week. The eosinophilic count reached its peak by the 2nd week. There was a significant inverse correlation between the mean eosinophilic count and the mean larval count. FAST-ELISA proved to be more sensitive than ELISA in early detection of infection, besides being a simple, fast and sensitive assay for antibody detection against T. spiralis. PMID- 12561905 TI - Study of factors affecting growth of Leishmania in cultures. AB - The rate of growth of Leishmania major and L. infantum in El-On's culture media supplemented with human, dog, rat and avian blood was studied in vitro. Rabbit blood was used as a control. The effect of culture with these types of blood on the infectivity of both Leishmania strains to albino mice was also studied. The results showed that a good yield of both L. major and L. infantum parasites can be obtained in culture by using avian blood as substitute for rabbit blood in El ON's medium. In addition, rat blood gave good results with L. infantum. The morphological forms of L. major and L. infantum on all types of blood supplemented media: elongated promastigotes, spindle promastigotes, paramastigotes and amastigoes were present all through the culture period with variable percentages. The infectivity to experimental animals was not affected by culture of both Leishmania strains on rabbit, human, rat, dog as well as avian blood supplemented media. PMID- 12561906 TI - Eimeria chiropteri n.sp. (Apicomplexa: Eimeriidae) from Saudi Arabian bat Pipistrellus kuhlii (Chiroptera: Vespertilionidae). AB - Oocysts of Eimeria chiropteri n.sp. are described from faeces of Pipistrellus kuhlii from Saudi Arabia. The sporulated oocyst of E. chiropteri is broadly ellipsoidal 23.5 x 20.6 (18.8 - 25.9 x 16.5 - 25.2) microm, and bilayered wall 1.2 (1.1 - 1.3) microm thick. Micropyle is absent, but a single, spherical polar body present. Oocyst residuum is present as a coarse granular sphere. Sporocyst is ovoid 10.8 x 7.5 (10.1 - 11.7 x 6.8 - 8.1). Stieda body is present, but sub and parastieda bodies are absent. Sporozoites are elongated laying head to tail in the sporocyst each with a large spherical refractile globule. E. chiropteri is the 4th species from the genus Eimeria detected in bats of the genus Pipistrellus. PMID- 12561907 TI - An enterotoxic activity of 18-20 kDa Cryptosporidium parvum secretory coproantigen (CCA 18-20 kDa). AB - An 18-20 kDa Cryptosporidum coproantigen (18-20 CCA), had been detected in the stool of infected humans and calves. A purified, electroeluted and concentrated 18-20 kDa antigen was tested in Ussing chamber, and electric parameters were tested before and after the addition of this antigen. A significant increase in the short circuit current (Isc) was detected. The enterotoxic effectof 18-20 kDa CCA, was time and dose dependent, heat labile, and Cl- dependent. The detected change in the short circuit current was not detected when the 18-20 kDa antigen wa incubated with its monospecific antibodies. These results indicate an enterotoxic activity of the 18-20 kDa antigen secreted from the released parasite and detected in stool of infected humans and calves. Additionally, they may help in developing an appropriate anti-secretory therapy for the intractable diarrhoea caused by cryptosporidiosis. PMID- 12561908 TI - Coccidian parasites as a cause of watery diarrhoea among protein energy malnourished and other immunocompromised Egyptian children. AB - The role of coccidian parasites in the pathogenesis of watery diarrhea was studied among children with protein energy malnutrition (PEM), immunocompromised due to causes other than PEM and immunocompetent diarrheic of matched age and sex, as controls. The results showed that the prevalence of infection was 15.48%, Cryptosporidium was the most prevalent and showed 14.19% (18.3%, 17.5% and 7.3% in PEM, immunocompromised and immunocompetent cases respectively). Cyclospora oocysts were detected only among 2 cases (1.29%) of PEM group. Isospora oocysts were not detected in any of the studied groups. Modified Ziehl-Neelsen (Z.N.) technique was found to be the most reliable technique for identification of coccidian protozoa infection in stool. Giardia lamblia cysts were found in 10.97% and Entamoeba histolytica in 5.16% of cases. Mixed infection (G. lamblia and E. histolytica) was found in 2.58% of the cases. The duration of diarrhoea was more prolonged in Cryptosporidium and Cyclospora infections among PEM and immunocompromised cases. Cryptosporidium is one of the important casuses of watery diarrhoea in infants and children in PEM and immunocompromised patients. Therefore, it is indicated to use modified Z.N. technique as a routine test for stool examination and immunocompromised patients must avoid contaminated water. PMID- 12561909 TI - Induction of some digestive enzymes in the midgut of the sandfly Phlebotomus langeroni after sugar and blood meals. AB - Changes associated with blood and sugar meal digestion in the sandfly, Phlebotomus langeroni were characterized. Different types of sugars: sucrose, glucose, melibiose, cellobiose, lactose, starch, fig fruits, honey dew and a mixture of sucrose and sugar sources were used for the sandfly feeding. Activities of glycosidases and proteases in the sandfly guts after blood and sugar meals were determined using the endpoint method. The results showed that glycosidases (alpha-glycosidase, beta-glycosidase, alpha-galactosidase, and beta galactosidase) are present in the sandfly midguts. No activities of the glycosidases (alpha-mannosidase and alpha-amylase) were detected in the sandfly gut. Proteases: trypsin and aminopeptidase showed activities in the sandfly midguts. It is concluded that the midgut glycosidase may play an important role in the vector-parasite interaction. Trypsin and aminopeptidase induction after a blood meal is controlled by a secretogogue mechanism which indirectly influences the outcome of the Leishmania parasite infection. PMID- 12561910 TI - Infection of two lymnaeid snails with Fasciola gigantica in Giza, a field study. AB - Population density of Lymnaea natalensis and L. columella and their natural infection with Fasciola gigantica were followed in two water bodies in Abu Rawash area, Giza Governorate over two years. L. natalensis snails were more abundant during December-February, while L. columella snails had high density during autumn. According to the snail size, we found that the reproduction of the two vector snails occurred in summer and early autumn. Fasciola was present in 0.6 and 2.0% of the examined L. natalensis and L. columella snails respectively. A high prevalence of F. gigantica in L. columella snails occurred in summer and early autumn, while their presence in L. natalensis was irregular. Naturally infected Lymnaea snails, with Fasciola, are usually of large size (> 10mm) of a given population sample. Other three types of trematode cercariae were found in both snail species; Xiphidiocercariae, Echinostomes and Amphistomes. PMID- 12561911 TI - Efficacy of BaySir-8514 and precocene II against the grey flesh fly Parasarcophaga dux (Thomson) (Diptera: Sarcophagidae). AB - Five doses (1, 0.5, 0.25, 0.125 and 0.0625) of the chitin inhibitor BaySir-8514, and anti-juvenoid precocene II were topically applied to the third larval instar of the grey flesh fly Parasarcophaga dux. Toxicity of both compounds were recorded against different developmental stages at the different dose levels. The lowest dose (0.0625) of both compounds caused a low toxicity percent (0.1). On the other hand, the highest dose (1mg) increased the larval, pupal and adult malformations after larval treatment by both compounds. Highest percentage of larval deformations was obtained by the maximal dose-level (1mg), which in turn did not lead to the highest percentage of adult deformations. Shortening of larval-pupal durations and adult longevities were observed. Sterilizing action of precocene II on P. dux could be remarkably detected by the effect of higher doses (0.5 and 1%) on the 3rd larval instar because no larvae were deposited by the emerging adult females. Doses of 0.125 and 0.0625 mg inhibited the female natality. PMID- 12561912 TI - Evaluation of ELISA kit hypodermosis in serodiagnosis of Przhevalskiana silenus in goats and Cephalopenia titillator in camels. AB - Myiasis is a problem of Medical and Veterinary importance worldwide. It affects human welfare both directly and indirectly. ELISA kit hypodermosis was evaluated in detecting antibodies against two oestrid larvae, Przhevalskiana silenus in goats and Cephalopina titillator in camels. Both animals were parasitologically proved to have oestrid myiasis infestation. Antibodies against P. silenus were detected in 38 out of 40 (95%) infested goats and negative results among the control group. With C. titillator, antibodies were detected in 39 out of 40 (97.5%) infested camels. But one of the control (10%) gave false seropositivity. ELISA kit hypodermosis diagnosed antibodies against P. silenus in goats and C. titillator in camels. Consequently, there are cross reactions between anti hypoderma antibodies and antibodies against both Przhevalskiana and Cephalopina. ELISA is confirmed to be an easy and economic tool in diagnosing myiasis infestation when direct examination is not a simple mean and/or for epidemiological studies. The whole results were discussed. Focusing on the clinical pictures and pathogenesis of human myiasis was included to clarify the zoonotic importance of myiasis in Egypt. PMID- 12561913 TI - Study of gamma-interferon in schistosomiasis mansoni, autoimmune diseases and schistosomal arthropathy. AB - IFN-gamma, a Th1 cytokine was quantitatively estimated using EASIA in the sera of patients with different stages of schistosomiasis mansoni autoimmune diseases and schistosomal arthropathy. Level of IFN-gamma was significantly elevated in all the test groups compared to the control group. The highest level of IFN-gamma observed was in RA and SLE, with insignificant statistical difference (P > 0.05) between them. Cases with early S. mansoni infection showed significant higher level (P < 0.05) of IFN-gamma compared to hepatosplenic schistosomiasis with and without ascites. A significant statistical difference (P < 0.05) concerning IFN gamma level was observed when comparing RA and SLE to different stages of S. mansoni infection and schistosomal arthropathy. This might lead to a conclusion that IFN-gamma estimation in sera of patients having joint complaint could be a good marker for differentiation between autoimmune collagen induced arthritis and schistosomiasis induced arthropathy. PMID- 12561914 TI - Value of estimating intraocular antibody production in diagnosis of typical and atypical lesions of ocular toxoplasmosis. AB - Serum and aqueous humor (AH) samples were collected from 45 patients: 20 with typically active or reactivated retinal lesions of Toxoplasma (Group I), 16 with atypical lesions (Group II) and 9 with old quiescent scars (Group III). Also, serum and AH samples were collected from 10 patients with chronic toxoplasmosis without any ocular manifestation (Group IV). T. gondii specific IgG, IgA and IgM antibodies were measured by ELISA in AH and serum and the intraocular (local) antibody production was determined by calculating Goldman-Witmer coefficient (G.W.C.). IgG antibodies were the only class detected in all sera of patients with ocular and nonocular toxoplasmosis. An intraocular IgG antibody synthesis was confirmed in 95% and 37.5% of patients with typical (Group I) and atypical (Group II) posterior uveitis respectively and in none of either patients with quiescent scars (Group III) or the ophthalmologically free patients (Group IV). As regard the typical active lesions, the sensitivity of the IgG assay (95%) was higher than that of IgA (60%) and IgM (5%) assays. Beside the conclusion that AH analysis to detect local antibody production is more reliable than the estimating of serum antibodies for the diagnosis of ocular toxoplasmosis, the detection of AH specific antibodies in 6 atypical cases, who were treated successfully by antitoxoplasmic therapy, represent a help to increase the number of uveitis cases in which specific treatment can be established. PMID- 12561915 TI - Mites infesting two migratory birds, Coturnix c. coturnix (quail or Simmaan) and Sturnus v. vulgaris (starling or zarzuur) with reference to avian zoonosis. AB - Birds are a marvelous group of creatures. Their beautiful coloration, singing, dancing and their attractive ways of life offer great pleasure to birdwatcher. Egypt is one of the most important countries of migratory birds. Not less than 300 species of birds visit Egypt annually from allover the world. The mite fauna of two migratory birds was studied in North Sinai Governorate and Suez Canal Zone. Thirty-one species of mites were recovered from quail and 39 from starling. Both types of birds serve hosts for 26 species of mites. Besides, five species were only recovered from quail and 13 species from starling. These totaled 44 species belonging to 30 families of three suborders (Mesostigmata, Trombidiformes and Sarcoptidiformes). The mite index on quail ranged between 1.0 to 5.0 in North Sinai G. and 1.0 to 17.0 in Suez Canal Z. The mite index on starling ranged between 1.0 to 4.75 in North Sinai G. and 1.0 to 4.5 in Suez Canal Z. Sixteen of the recovered species of mites were not recorded before on the Egyptian resident birds (house sparrow and laughing dove). The medical and veterinary importance of avian zoonosis was discussed. It is hoped to stimulate the awareness to migratory birds as reservoir hosts for microorganisms and parasites from allover the world. Besides, there is an urgent need to protect the resident and the visiting birds. PMID- 12561916 TI - The potential use of N,N-diethyl-m-toluamide (DEET) as a prophylactic agent in the control of schistosomiasis. AB - DEET (N, N-diethyl-m-toluamide) is one of the reliable and most widely used insect repellents. This work was planned to study the effect of DEET on Schistosoma mansoni cercariae viability (in vitro) and skin penetration and migration (in vivo). DEET concentrations of 30%, 15% and 7.5% were highly efficient in killing all cercariae in vitro within 30, 60 and 240 minutes respectively. Two concentrations of DEET (15%) and (7.5%) were tested separately for their antipenetrant and protective effects in mice by immersing their tails in the drug and then exposed to cercariae, 30 minutes later. DEET antischistosomal activity was assessed by the number of non penetrating cercariae, worm burden, egg count/gram liver and number of liver granulomata. The maximum prophylactic effect was recorded using DEET (15%) where the percentage of resistance reached 100% compared to 92% using DEET (7.5%). DEET was highly effective in preventing penetration of cercariae as well as its further migration to liver which is proved histopathologically in the liver. The persistence of the antipenetrant and protective effects of DEET over several days post application and following different water wash durations was examined. Treatment of mice tails with DEET (15%) and (7.5%), three days prior to infection, showed significant levels of resistance of 94.8% and 82.7% respectively. DEET (15%) provided 100% and 97.9% resistance even when treated tails were washed for 1 and 2 hours respectively. PMID- 12561917 TI - Immunofluorescent detection of both Giardia lamblia and Cryptosporidium parvum using anti-Cryptosporidium oocyst antibodies. AB - Giardia lamblia and Cryptosporidium parvum are two protozoal parasites proved to have a major role in gastroenteritis in humans. Both are documented to coexist in many waterborne parasitic transmission, as well as in outbreaks. In the present work, a polyspecific anti-Cryptosporidium oocyst antibodies were used for simultaneous detection of both parasites in human stool. Known positive formalinized human stool specimens of Giardia sp. (n = 10), Cryptosporidium sp. (n = 7), mixed infection (n = 3), and negative specimens (n = 20), were tested using direct fluorescent technique against the developed antibodies. All positive stool samples for Cryptosporidium and 9 out of 10 Giardia samples, or each alone showed fluorescence with variable intensities, while no negative sample harboured other parasites had fluorescence. This newly used polyspecific antibodies offer the advantages of screening large number of patients, particularly in outbreaks. Additionally, it represents a cheaper alternative for the most sophisticated and costly immunoassay kits using the monoclonal antibodies, with more or less the same diagnostic potentials. PMID- 12561918 TI - Human nasopharyngeal linguatuliasis (Pentasomida) caused by Linguatula serrata. AB - A human nasopharyngeal linguatuliasis was reported for the second time in Egypt. The patient (20 years old male) was presented with main conspicuous complaints, fever, urticaria (face and neck), coughing, vomiting and passage of small (less than 1 cm. in length) worm-like structures in his nasal discharge and vomitus. Symptomatic treatment was given followed by a single dose of praziquantel after identification of the causative parasite. Human linguatuliasis (pentastomiasis) was discussed. PMID- 12561920 TI - Detection of hydatid antigen in human cyst fluid by reverse latex agglutination test. AB - Diagnosis of cystic echinococcosis (CE) has, sometimes, proved to be of some difficulties whether by serological, radiological, or even by direct microscopic examination of cyst fluid. Therefore, with an aim to apply and assess another method of diagnosis, of pre - intra - or post-operative application, the present work had studied the use of reverse latex agglutination (RLA) test in the diagnosis of CE, as a test that could directly detect hydatid antigen in human cyst fluid samples. The results were reliable when compared with the direct microscopic examination of cyst fluid samples. Also, it was more reliable than indirect haemagglutination test. It showed 100% sensitivity. No false positive reaction was observed with samples from cystic tumours, with a resultant specificity of 100%. Moreover, the test is easy to perform with a visually interpreted results within 2-3 minutes. The reagents are inexpensive, stable and easily available. These merits make the RLA test suitable for the diagnosis of CE, particularly in suspected cases with seronegative results or cases with sterile cysts. PMID- 12561919 TI - Effect of deferoxamine alone and combined with pyrimethamine on acute toxoplasmosis in mice. AB - The standard regimen of treatment for toxoplasmosis is pyrimethamine with sulfadiazine. However, it is not suitable in some conditions, and nontolerable in AIDS patients. Deferoxamine (DFO), an iron chelator, is well tolerated clinically in transfusion induced - iron overload. The present study had shown that DFO is a promising drug against acute toxoplasmosis in mice. Three doses of 200, 300 and 400 mg/kg DFO were used either alone or in combination with pyrimethamine. Alone, it was effective in a dose- related response with the resultant of 70% protection of infected mice. When DFO was combined with a low minimally effective dose of pyrimethamine, a 100% protection was recorded with the prolongation in duration of survival of mice. Different possible mechanisms of action of DFO against Toxoplasma gondii were discussed. PMID- 12561921 TI - Human cystic echinococcosis: diagnostic value of different antigenic fractions of hydatid cyst fluid with different specific immunoglobulin G subclasses by enzyme linked immunoelectrotransfer blot. AB - The present study intended to evaluate the usefulness of immunoblot analysis of hydatid cyst fluid (HCF) for diagnostic verification of human cystic echinococcosis (CE). HCF of camel origin was resolved by SDS-PAGE gel electrophoresis and transblotted on nitrocellulose membrane. Immunoglobulin G (IgG) subclass-specific antibody responses in 25 sera from surgically confirmed CE cases, 44 persons with other parasitic infections and 20 normal controls were analyzed. Total IgG and IgG subclass1-4 in CE sera preferentially recognized several polypeptide bands in the range of 14-200 kDa. The most predominant band recognized by total IgG antibodies was at 21 kDa (sensitivity, 96%; specificity, 98.5%; J index, 94.5%), by IgG1 at 38 kDa (sensitivity, 92%; specificity, 100%; J index, 92%) and by IgG3 at 60 kDa (sensitivity, 96%; specificity, 100%; J index 96%). Sera from the normal controls did not recognize any of these polypeptides. These data suggest that detection of any of these polypeptides bands could be used for confirmation of human cystic echinococcosis in Egypt. PMID- 12561922 TI - A new host and locality records of the two trematodes Gymnotergestia chaetodipteri and Opechona sardinellae described by Nahhas and Cable in 1964, with review of the two genera. AB - The two trematode genera Gymnotergestia Nahhas and Cable, 1964 and Opechona Looss, 1907 were reviewed and discussed. Gymnotergestia chaetodipteri Nahhas and Cable, 1964 and Opechona sardinellae Nahhas and Cable, 1964 were redescribed from Serranus scriba and Labrus bergylata respectively from Libya. This represents a new host record and new locality. A comparison between the original description and the present work was presented. PMID- 12561923 TI - Scanning electron microscopical studies on cercariae and metacercariae of Fasciola gigantica. AB - The surface topography of Fasciola gigantica cercariae and metacercariae was studied by scanning electron microscopy. The head of F. gigantica cercaria is covered with several small knobs and its tail is provided with two lateral folds, fused ventrally, near the distal end of the tail. The oral sucker is smaller than the ventral one and possesses a characteristic surface structure. The outermost layer (layer I) of the outer cyst wall of the metacercaria is roughened with irregular furrows. The inner surface of the outer cyst wall (layer II) is more homogeneous and nearly smooth. The outermost layer of the inner cyst wall (layer III) is smooth and lacks any furrows or tubercles. The differences between F. gigantica and F. hepatica cercariae and metacercariae are discussed. PMID- 12561924 TI - An enzyme-linked immunoelectrotransfer blot assay for diagnosis of human cystic echinococcosis. AB - An enzyme-linked immunoelectrotransfer blot (EITB) test was assessed for diagnosis of 47 pulmonary cystic echinococcosis (CE) cases admitted to Ain Shams University Hospitals for Surgery. Diagnosis of these cases was established on clinical examination, X-ray, sonography and indirect hemagglutination (IHA) test, which was negative in four cases (sensitivity 91.5%). Sera from patients with other parasitic infections, carcinomas or normal sera were used as controls. Human and Camel hydatid cyst fluids were used as antigens after separation and characterization of their antigenic components using 12.5% SDS-PAGE under reducing conditions. Six molecular weight antigens with molecular masses approximately 7, 20, 28, 35 and 127 kDa were of diagnostic importance. They were strongly recognized by sera of all CE patients specially with camel hydatid cyst fluid (HCF) giving a 100% sensitivity to the EITB test. Sera of patients with other parasitic infections as well as carcinomas and normal control sera couldn't recognize any of the above antigens and therefore were negative for the test. This results in 100% specificity of the EITB test. These data support the concept that EITB using camel hydatid fluid is a good diagnostic test for cystic echinococcosis. PMID- 12561925 TI - Effect of dual infections of Escherichia coli and pure caecal Eimeria sp. in broiler chickens. AB - The interactive effect of Escherichia coli and pure caecal Eimeria spp. in broiler chickens was studied. Single and dual orally inoculations were assessed in six groups of 20 chicks each. Neither mortalities nor clinical signs were recorded on inoculated groups or control. There was a significant decrease in body weights in groups infected, either with E. coli or caecal Eimeria spp. or with both pathogens. The total oocyst output and caecal lesion score were high and severe in chicks infected with caecal Eimeria sp. alone or with E. coli prior to caecal Eimeria infection. Low oocyst output and moderate caecal lesion score were noticed in chicks inoculated with E. coli prior to or together with caecal Eimeria infection. The results suggested a synergistic action between E. coli and caecal Eimeria sp. in producing gross lesions. The electrophoretic distribution of serum proteins decreased in all chicks compared with control. The changes in serum protein fractions differed in inoculated groups and all protein components decreased quantitatively compared with controls. It was also shown that E. coli did not always invade the blood stream. PMID- 12561927 TI - Evaluation of five stains in diagnosing human intestinal coccidiosis. AB - Three hundreds cases over the year 1998 complaining of diarrhoea were examined. The stools were examined by the traditional diagnostic methods and confirmed to be free from intestinal parasites. Cryptosporidium parvum, Isospora belli and Cyclospora cayetanensis were detected by using different types of faecal stains namely modified Ziehl-Neelsen, Kinyoun acid-fast, Auramine-rhodamine, Gomori's trichrome and Giemsa. The number of positive cases were 87 C. parvum (29%), 5 cases L. belli (1.7%) and 12 cases C. cayetanensis (4%). This study showed that the sensitivity of modified Ziehl-Neelsen and Kinyoun Acid-fast were very high (100%) in comparison with the other stains. PMID- 12561926 TI - Evaluation of the role of Ancylostoma caninum in humans as a cause of acute and recurrent abdominal pain. AB - Ancylostoma caninum is responsible for cases with eosinophilic enteritis (EE) and unexplained abdominal pain with peripheral eosinophilia in man. Ninety-five patients with obscure acute or recurrent abdominal pain and ten asymptomatic healthy parasite free were subjected to thorough history taking, clinical examination, sonography, routine laboratory investigations and serotesting by IgG ELISA to detect antibodies to excretory/secretory (ES) antigens of adult A. caninum and by IgG and IgG4 Western blot (W.B.) to detect antibodies to Ac68 antigen. Eleven male patients (11.6%) (5 with acute abdomen, 3 diagnosed as appendicitis and 3 had recurrent mild to moderate abdominal pain) fulfilled the criteria of case definition of human enteric infection with A. caninum (G.I). The study also detected human hookworm infection in 14 patients (G.IIb) other parasites in 34 patients (GIIc) and 36 patients had no parasites (G.IIa). Although 3 patients from group I were diagnosed as appendicitis and were dealt with surgically, the pain recurred and mebendazole only put an end to the patient's complaints. The obtained appendices of these operated cases showed marked eosinophilic infiltration but no adult canine hookworms were detected. IgG ELISA was positive in 72.7%, 8.3%, 100%, 23.5% and 0% in groups and control respectively. IgG and IgG4 W.B. did not increase the sensitivity but IgG4 W.B. elevated specificity to 100% excluding those with HH infection (Group Iib) who showed 100% cross-reactions. Stool analysis was the only differentiation between these two types of hookworms. These findings confirmed the presence of human enteric infection with A. caninum as clinical entity in the study community and referred to its value in differential diagnosis of the obscure abdominal pain. PMID- 12561928 TI - A four years old girl with phthiriasis pubis infestation. AB - A four years old girl had Phthirus pubis infestation in the scalp, eyelids, eyelashes, neck and shoulders. She had fever, severe itching, allergic reaction and anxiety. The girl was successfully treated with 2.5% permethrin cream. The risk of lice infestation as a community health problem was discussed. PMID- 12561929 TI - Study on rodents infestation in Alexandria and prevalence of Trichinella spiralis infection among them. AB - Rodents were monthly collected from the six districts of Alexandria all year round using live traps. The 1025 collected rodents were divided according to species, gender, age, and weight El-Amryia and El-Montaza were the most infested districts, while West, East, Middle and El-Gomrok Districts were the lowest infested ones. Rallus norvegicus was the highest abundant species in the city (41.9%) followed by R. rattus (37.2%), then Mus musculus (20.9%). There was no difference in abundance of rodents during summer compared with winter. Meanwhile, males represented 57.6% of the total collected rodents. Most of the collected rodents were < or = 2 months old (63.8%) or light weighed (61.6%) however, number of caught rodents became less and less as age or weight increased. Concerning El Amryia and El-Montaza Districts where abattoirs are located, number of collected rodents from and around the abattoirs represented 70.4% of the total collected rodents compared with 29.6% collected from and around the houses of the same districts. The total prevalence of T. spiralis infection among collected rodents using digestion technique was 13.3%. El-Amryia, El-Montaza, West and El-Gomrok Districts were, in order, the only infected districts. Neither gender nor season of collection has influence on the prevalence of infection. The present study proved the role of abattoirs in the prevalence of T. spiralis infection. However, high prevalence of T. spiralis infection was detected among higher age or higher body weight groups of rodents. Sensitivity of the digestion diagnostic technique is proved compared with compressorium diagnostic technique. Nevertheless, diaphragm was the most sensitive site for detection of light infection in all studied rodents followed by thigh then tongue; expressed as number of larvae per gram tissue. PMID- 12561930 TI - Detection of Fasciola-specific excretory/ secretory (E/S) protein fraction band (49.5 kDa) and its utilization in diagnosis of early fascioliasis using different diagnostic techniques. AB - The objective of the present work is to evaluate Fasciola E/S antigens for diagnosis of early fascioliasis utilizing different diagnostic techniques. Using enzyme-linked immunoelectro-transfer blot (EITB), Fasciola-specific E/ S protein fraction band (49.5 kDa) was determined and electroeluted. The mono-specific antibodies against this specific fraction band were prepared by immunizing pathogen-free rabbit. Assessment of the prepared mono-specific antibodies in diagnosis of human fascioliasis was performed through the detection of E/S copro antigens by enzyme-linked immunosorbent assay (ELISA) in stool eluates obtained from patients with confirmed fascioliasis, other parasites as well as from other healthy individuals. Serum samples were collected and tested to detect serum antibodies against Fasciola E/S antigen using EITB and counter immunoelectrophoresis (CIEP). Analysis of Fasciola adult worm E/S products by SDS/PAGE revealed a number of bands, the molecular weight (MW) of which ranged from 14-200 kDa; with three major bands (27.5, 32.5 and 55 kDa). Fasciola EIS 49.5 kDa protein fraction proved to be specific to F. gigantica. Cross reaction with S. mansoni was observed at higher MW (110-120 kDa). The sensitivity, specificity and diagnostic accuracy of EITB were 45.2%, 100% and 70.7%, respectively, while those of CIEP were 38.7%, 100% and 67.2%, respectively. ELISA technique using mono-specific anti-49.5 kDa to detect copro-antigens proved to be practical and reliable. It showed higher sensitivity (91.4%) and higher diagnostic accuracy (91.8%), while the specificity was 92.3%. In addition, ELISA had higher negative predictive value (88.9%) and fair positive predictive value (94.1%). PMID- 12561931 TI - Quantitative inhibition ELISA for diagnosis of schistosomiasis using human IgG labeled with fluorescein and anti-fluorescein/peroxidase. AB - Usually mouse monoclonal antibodies are used in inhibition assays for antibody determination. Interference may occur in these inhibition assays due to presence of naturally occurring anti-mouse antibodies in some human serum samples. To avoid such interference, human IgG isolated from a pool of serum samples of S. mansoni patients and highly positive for IgG against S. mansoni soluble egg antigen (SEA) was used in inhibition ELISA for diagnosis of S. mansoni infection. The assay was based on inhibition of binding of human IgG labeled with fluorescein to S. mansoni SEA coating microtitration plates by tested serum samples. Plates were washed and labeled human IgG reacted with SEA was linked to peroxidase enzyme by incubation with anti-fluorescein/peroxidase conjugate. The assay showed 90% sensitivity and 96.3% specificity. The level of inhibition in ELISA showed highly significant positive correlation with stool egg output (Kandall's tau b = 0.512, P < 0.001). To make the assay quantitative, serial dilutions of the highly positive human serum pool, used for preparation of human IgG, were applied in each plate and concentration of anti-SEA antibodies in serum samples tested was calculated from a 4-parameters logistic curve equation. The highly positive serum pool used as a standard was considered to contain one million arbitrary units of immunoglobulins against S. mansoni SEA. Human IgG is expected to be more practical in inhibition assays than mouse monoclonal antibodies due to elimination of interference caused by naturally occurring human anti-mouse antibodies. Also, large amount of human IgG could be purfied from remnants of serum samples highly positive for the proposed antibodies. A higher specificity and sensitivity could be obtained if IgG is isolated by affinity purification instead of ammonium sulphate precipitation. In conclusion, human IgG isolated from highly positive serum samples could be used in sensitive and specific diagnostic antibody determination inhibition assays for diagnosis of infectious and autoimmune diseases. PMID- 12561932 TI - The insecticidal activity of Cyanobacteria against four insects, two of medical importance and two agricultural pests with reference to the action on albino mice. AB - Acute lethal toxicity of Cyanobacteria (blue-green algae) was investigated against 3rd larval stages of Musca domestica, and Culex pipiens, and 4th larval stages of Spodoptera littoralis and Agrotis ipsilon. The LD50s were 1.94, 2.33, 7.59 and 9.10 microg for M. domestica, C. pipiens, S. littoralis and A. ipsilon respectively. The slope functions were 4,045 +/- 0.332 (M. domestica), 4.122 +/- 0.336 (C. pipiens), 4.15 +/- 0.35 (S. littoralis) and 3.72 +/- 0.34 (A. ipsilon). These LD50s markedly affected the larval, pupal and adult time of durations as well as suppressed the oviposition of the survivor adults. The LD50 didn't show any marked effect as serum acetyl cholinesterase and gave nearly the same level of AchE activity. No doubt, Cyanobacteria are safe and promising agent for insect control. PMID- 12561933 TI - Opportunistic intestinal protozoal infections in immunocompromised children. AB - Hundred immunocompromised children and 100 house contact controls were chosen. Patients included: 52 nephrotic syndrome children receiving corticosteroids for more than one month (age 5.28 +/- 2.32 years), 14 protein-calorie malnutrition (PCM) patients (8 cases of marasmus aged 6 +/- 2.27 months and 6 cases of marasmic kwashiorkor aged 1.39 +/- 0.88 years) and 34 lymphomas patients (22 cases of Hodgkin's disease and 12 cases of non-Hodgkin's lymphoma; age 4.5 +/- 3.54 years). Examination of concentrated stool was done using iodine stain of fresh mounts and modified Ziehl-Neelsen (cold acid-fast) to fixed smears. T-cell subsets were counted after staining with mouse monoclonal antibodies against CD4 and CD8 labeled with fluorescein. Both nephrotic syndrome and lymphomas groups showed affection of cellular immunity in the form of significant decrease in T helper and H/S ratio and significant increase in suppressor T-cell subsets. Giardia lamblia, Entamoeba histolytica, Cryptosporidium parvum and Blastocystis hominis were the most frequent in patients group and were significantly more prevalent among patients than controls. No significant difference in the prevalence of Entamoeba coli and Chylomastix mesnili between the two groups. C. parvum infection were strictly confined to groups with T-cell subsets abnormalities i.e. nephrotic syndrome and lymphomas groups. PMID- 12561934 TI - Reciprocal heterologous protection between Trichinella spiralis and Toxoplasma gondii concurrently present in experimental murine models. AB - Mixed parasitic infections have been long seen as a double impact on the host. In this study a different view of polyparasitism is demonstrated. The influence of the immunological environments created by two biologically different parasites on the pathogenesis of each other was evaluated. Swiss albino mice were sequentially infected with the intracellular protozoan Toxoplasma gondii (acute and latent), which elicits a T-helper 1 (Th1)-polarized immune response and the helminth parasite Trichinella spiralis, whose infection is predominated by a Th2 response. The results show a significant heterologous protection from one parasite towards the other. There was a highly significant lower Trichinella muscle larvae burden in mixed infection group compared to single Trichinella infection, in spite of delayed intestinal adult worm expulsion in the mixed infection group. A highly significant lower burden of T. gondii brain cysts in mixed infection compared to single latent Toxoplasma infection was also demonstrated. Concerning the anti Toxoplasma antibody response, there was a significant lower levels in the latent Toxo-Trich. group compared to the group of latent toxoplasmosis only. These significant lower antibody titers were reproducible by two different assays; dye test and direct agglutination test. PMID- 12561935 TI - Studies on the liver fluke Fasciola in Egypt: I--Morphological and morphometrical studies. AB - A comparative scanning electron microscopy and morphometrical study of the two geographically isolated species of Fasciola (F. heptica European isolate and F. gigantica from Egypt) were studied in order to clarify their genetic relationships and specific identification. Although the present study has revealed that most of the diagnostic morphological and morphometrical criteria in the two species are highly variable, the position of the ventral sucker relative to the whole body length produced the most significant differentiating criterion, in addition to the presence of markedly larger tegumental papillae on the ventral surface of F. gigiantica. PMID- 12561936 TI - Pathological alterations in the ovaries of Culex pipiens induced by fixed oil extracts from Thevetia peruvine, Datura stramonium and Acacia sp. AB - The fixed oil extracts from Thevetia peruvine, Datura stramonium and Acacia sp. were tested on Culex pipiens larvae. The estimated sublethal concentrations (LC50) were used in the present studies. The reproductive potential of females and the histochemistry of their ovaries were determined. The results indicated that oil treatments of larvae caused drastic changes in reproduction potential of female mosquitoes including the ovarian development in the first gonotrophic cycle, fecundity and fertility of treated females. Histological and histochemical studies revealed clear changes in the oocytes of females derived from treated larvae. Treatment with the oils reduced the vitellogenesis, synthesis of proteins, carbohydrates, lipid contents and DNA and RNA materials. The reproduction was always significant and may reflect the shortage in reproductive potential, the state of being disturbed. Generally, these fixed oils merit further investigations as a potential suppressant of Culex pipiens because their abilities to kill the early stages, inhibit growth and development of ovaries and reduce the reproductive potential of adult females. The three oils have variable activities but they are similar in their mode and site of actions. PMID- 12561937 TI - Production of pro-inflammatory cytokines (GM-CSF, IL-8 and IL-6) by monocytes from fasciolosis patients. AB - The production of pro-inflammatory cytokines by monocytes in vitro has been measured in eight patients with acute fasciolosis and 15 patients in the chronic stage of the disease, before and after stimulation by excretory/secretory Fasciola antigen. Results were compared with those of a control group of 12 individuals. The monocytes from patients with acute fasciolosis produced significantly higher levels of GM-CSF, IL-8 and IL-6 as compared to controls. With chronicity, the production of these cytokines was decreased as compared to the acute stage probably due to decreased antigen level in blood. Stimulation of monocytes of healthy control with E/S Fasciola antigen was accompanied with a markedly increased production of pro-inflammatory cytokines, while monocytes from patients with acute or chronic fasciolosis revealed minimal increase in production. This denoted the importance of E/S Fasciola antigen as an activator of monocytes. A second exposure to the same antigen was accompanied with a limited response. PMID- 12561938 TI - Parasitological effects of simultaneous infections induced by Schistosoma mansoni and aflatoxin B1 in Syrian golden hamsters. AB - To determine the effect of S. mansoni and aflatoxin B1, 120 hamsters were divided into six groups of 20 each as follows: group I, S. mansoni only, group II, aflatoxin B1 only, group III, S. mansoni then aflatoxin B1 eight week, group IV, aflatoxin B1 then infected with S. mansoni six weeks, later, group V, aflatoxin B1 and S. mansoni simultaneously, group VI control. Loss of body, liver, and spleen weight was more prominent in groups IV and V than in other four groups. The higher mortality rate was in group III. Animals treated with aflatoxin followed by S. mansoni infection appeared to have a less deteriorating effect on the liver (group IV) than group III treated with by S. mansoni first followed by aflatoxin treatment. No morphological abnormalities were detected in the worms including testicular changes in males but a significant number of females was immature even in copula (P < 0.01) in groups III, IV, and V. The average number of S. mansoni eggs was less in groups III, IV, and V in comparison to group I. No abnormalities were detected in the eggs for groups infected with S. mansoni. The diameters of granulomas around eggs, were large on an average in groups III, IV and V as compared with groups I and II. PMID- 12561939 TI - Diagnosis of symptomatic and asymptomatic Trichomonas vaginalis infection by applying one tube nested PCR to vaginal discharge. AB - One tube nested-PCR targeting a species-specific Tv-E650 repeat family of T. vaginalis genome, was applied to vaginal discharge specimen to diagnose symptomatic and asymptomatic trichomoniasis. The test was compared with axenic culture examination, wet mount preparation and Papanicolaou stained smears. Four hundred and fifty cases symptomatic and symptomatic were collected over two years. Out of 290 symptomatic women with cervicovaginitis and 160 asymptomatic women, 35 were culture positive for trichomoniasis. All culture positive specimens were PCR-positive giving a single product at 290 bp by agarose gel electrophoresis and recording 100% sensitivity, similar to culture examination. Among the 35 culture positive specimens, 12 were positive by wet mount and 21 were positive by Pap smear giving a 34.2% and 60% sensitivity, respectively. The standard and boiling DNA extraction methods were equally reliable but the latter was more simple, rapid and cheap. No specimens negative by PCR for trichomoniasis were positive by culture, wet mount or Pap smear. Moreover, specimens from cases with cervicovaginitis of non-trichomonal origin were negative by PCR. All samples of extracted DNA of T. vaginalis from positive culture tubes, used as positive controls, were also PCR positive at the expected product in gel, while no PCR product was detected with the negative DNA control including Chylamidia trachomatis and human DNA. It was found that the one-tube nested-PCR targeting the Tv-E650 repeat family of T. vaginalis is a simple, rapid sensitive and specific accurate method for diagnosis of symptomatic and asymptomatic vaginal trichomoniasis when applied to vaginal discharge. PMID- 12561940 TI - Metabolic programming by nutrition during early development. AB - Incidence of obesity and diabetes is increasing at an alarming rate not only among the populations of the affluent nations but also amongst the populations of the developing nations. Understanding the mechanisms that cause the onset of these pathological conditions is a requisite to effectively tackling this problem. In this context the role of early nutritional experiences as a causative factor is being extensively investigated. This article briefly reviews the field of metabolic programming vis-a-vis an altered nutritional milieu during perinatal period and consequent adaptive metabolic patterning and metabolic imprinting in adult and/or consequent offspring. PMID- 12561941 TI - Mechanism of plant growth promotion by rhizobacteria. AB - Plant growth results from interaction of roots and shoots with the environment. The environment for roots is the soil or planting medium which provide structural support as well as water and nutrients to the plant. Roots also support the growth and functions of a complex of microorganisms that can have a profound effect on the growth anti survival of plants. These microorganisms constitute rhizosphere microflora and can be categorized as deleterious, beneficial, or neutral with respect to root/plant health. Beneficial interactions between roots and microbes do occur in rhizosphere and can be enhanced. Increased plant growth and crop yield can be obtained upon inoculating seeds or roots with certain specific root-colonizing bacteria- 'plant growth promoting rhizobacteria'. In this review, we discuss the mechanisms by which plant growth promoting rhizobacteria may stimulate plant growth. PMID- 12561942 TI - Active immunization against riboflavin carrier protein results in peri implantation embryonic loss leading to pregnancy termination in rats: use of alternate adjuvants. AB - To investigate the mechanism of pregnancy termination following immuno neutralization of riboflavin carrier protein (RCP) and to use acceptable adjuvants, we actively immunized female rats with reduced and carboxymethylated RCP (RCM-RCP) using various adjuvants (during primary immunization) such as sodium phthalylated lipopolysaccharide (SPLPS), purified S. typhi outer membrane proteins (porins) and a combination of them. Rats (5-14 per group) were immunized with alugel adsorbed RCM-RCP (100 microg/dose) either alone or with SPLPS or porins or SPLPS+porins. Control animals received RCM-RCP emulsified with Fruend's completelincomplete adjuvants (FCA/FIA). All animals received five boosters at intervals of 21 days. The lowest (4 X 10(-3)) and the highest (> 70 X 10(-3)) anti-RCM-RCP antibody titers were observed in alugel adsorbed-RCM-RCP group and control groups, respectively. Immunized animals showed reduced fertility following 3rd, 4th and 5th boosters. Reduction in fertility was 30-60% in alugel adsorbed RCM-RCP group, 90-100% in FCA-RCM-RCP group and 80-90% in SPLPS+porins group. Fertility reduction was not strictly correlatable with the serum antibody titers. RCP-specific IgG could be localized in the uterine endometrial glands and luminal epithelial cells in the immunized animals. Animals in the FCA/FIA group showed abnormal implantation/resorption sites and their histological sections showed degenerated embryos. But, day 5 preimplantation embryos were normal. These results show that (a) SPLPS+porins can be used as adjuvants in place of FCA/FIA for active immunization against RCM-RCP and (b) early termination of pregnancy in the immunized animals is due largely to the failure of normal embryo implantation. PMID- 12561943 TI - GnRH and / or testosterone induced changes in reproductive activities during nonbreeding season in Calotes versicolor (Daud.). AB - Administration of Gonadotrophin releasing hormone (GnRH) to male C versicolor during nonbreeding season increases the weight of testis;diameter of testis, seminiferous tubule, Sertoli and Leydig cell nuclei. It also activates the spermatogenic process. Increase in the weight of epididymis and lowered cholesterol level of testis indicate androgen production. Treatment of tesotsterone along with GnRH further enhances the activities of testis as a few spermatozoa appeared in the lumen of seminiferous tubule along with increase in other spermatogenic elements. It may be concluded that the exogenous GnRH can induce reproductive activities during nonbreeding season when the environmental conditions are unfavourable. Testosterone administration has the additive effect on these activities. PMID- 12561944 TI - Antioxidant activity of tannoid principles of Emblica officinalis (amla) in chronic stress induced changes in rat brain. AB - Effect of tannoid principles emblicanin A, emblicanin B, punigluconin, and pedunculagin of E. officinalis was assessed on chronic unpredictable footshock induced stress-induced perturbations in oxidative free radical scavanging enzymes in rat brain frontal cortex and striatum. Chronic stress, administered over a period of 21 days, induced significant increase in rat brain frontal cortical and striatal superoxide dismutase (SOD) activity, concomitant with significant reduction in catalase (CAT) and glutathione peroxidase (GPX) activity. The changes in the enzyme activities was accompanied by an increase in lipid peroxidation, in terms of augmented thiobarbituric acid-reactive products. Administration of Emblica tannoids (10 and 20 mg, po) for 21 days, concomitant with the stress procedure, induced a dose-related alteration in the stress effects. Thus, a tendency towards normalization of the activities of SOD, CAT and GPX was noted in both the brain areas, together, with reduction in lipid peroxidation. The results indicate that the reported antistress rasayana activity of E. officinalis may be, at least partly due to its tendency to normalize stress induced perturbations in oxidative free radical scavenging activity, in view of the postulate that several stress-induced diseases, including the process of aging, may be related to accumulation of oxidative free radicals in different tissues. PMID- 12561945 TI - Phenyldiguanide activates cardiac receptors to produce responses by involving three different efferent pathways in anaesthetized rats. AB - The present study was undertaken to determine the afferent and efferent pathways involved in the phenyldiguanide (PDG)-induced reflex response in rats. Intravenous (iv) injection of PDG (10 microg/kg), produced hypotension, bradycardia and apnea over a period of time. Bilateral vagotomy abolished the PDG induced reflex changes. Atropine (2 mg/kg; iv) blocked only the bradycardiac response produced by PDG, while prazosin (0.5 mg/kg; iv) blocked the hypotensive response, and bilateral vagotomy in these animals abolished the apneic response. In separate series of experiments, intrapericardial injection of lignocaine abolished the hypotensive and bradycardiac responses evoked by PDG in artificially ventilated rats. The results reveal that the PDG-induced reflex is mediated through vagal afferents originating from the heart and efferents involve three different pathways. The bradycardiac response was through the muscarinic receptors, the hypotension is mediated through alpha1 adrenoceptors and the apnea presumably through the spinal motoneurones supplying the respiratory muscles. PMID- 12561946 TI - Poly (DL-lactide-co-glycolide) microparticles as carriers for antimycobacterial drug rifampicin. AB - Poly (DL-lactide-co-glycolide) polymers were investigated as carriers for the first line antitubercular drug rifampicin. Different formulations of PLG microparticles viz. porous, non porous and hardened exhibited sustained release of rifampicin up to 7 weeks in vitro. However, hardened PLG microparticles exhibited the most sustained release in vivo in different organs up to 6 weeks. In case of free rifampicin, release was detected in vivo only up to 48 hr. In addition, no hepatotoxicity was observed on a biochemical basis (levels of SGPT, ALP and total bilirubin) in comparison to control animals. Taken together, these results suggest that polymer encapsulated antitubercular drug rifampicin may serve as an ideal therapeutic approach for treatment of tuberculous infections. PMID- 12561947 TI - Role of volatile oil pretreatment and skin cholesterol on permeation of ion paired diclofenac sodium. AB - This study was designed to investigate the influence of volatile oil pretreated skin on in vitro permeation from films containing ionized and dodecylamine ion paired diclofenac sodium (DS). The involvement of skin cholesterol was investigated to determine its possible role in enhancing the permeation of ion paired DS. Cardamom oil produced the maximum (10 x) in vitro permeation enhancement for ion-paired DS. The carrageenan induced rat paw oedema reduction (up to 12 hr) by cardamom oil was comparable to that of diclofenac injection (s c). Leaching of cholesterol from excised skin in addition to increased partition coefficient following volatile oil skin pretreatment appears to be responsible for in vitro permeation enhancement of DS. Whereas, a mild barrier perturbation effect due to altered cholesterol levels following pretreatment with volatile oils appears to increase the permeation of ion-paired DS across viable skin, thereby producing significant reduction of carrageenan induced paw oedema. PMID- 12561949 TI - Ethinyl estradiol: its interaction on blood-lipid. AB - Lipophilicity (log P) of the drug plays an important role when drug reaches in the critical reaction site, i.e., active site cum receptors where the major constituent is lipid moieties. The drug molecule may be responsible for altering the lipid constituents, which is measured in terms of phosphorus content and can be explained by their fatty acid changes that are linked with biological effect of the drug. Having considered the lipophilicity of ethinyl estradiol (log P = 3.67), its interactions with the whole lipid of goat blood have been investigated along with fatty acid changes and lipid peroxidation phenomena. There was significant loss of phosphorus content of phospholipid and change of fatty acid constituents of whole lipid. This may be ascribed to binding affinity of ethinyl estradiol with lipid constituents in blood. Lipid binding potential of the drug may have role in its therapeutic effect. The peroxidation induced by drug has been quantitatively measured along with its suppression by using antioxidant. The results reveal that ethinyl estradiol caused significant extent of lipid peroxidation. Ascorbic acid, a promising antioxidant could significantly reduce drug induced lipid peroxidation. PMID- 12561948 TI - Pharmacokinetic study of niosome encapsulated insulin. AB - Pharmacokinetic profile and hypoglycemic effect, after intraperitoneal injection of insulin and insulin encapsulated in niosomes were determined in diabetic rats. Niosomes (non-ionic surfactant vesicles) of different doses and different lipid compositions were prepared by lipid layer hydration method. Plasma samples were collected at specified time intervals and plasma concentration of insulin was determined by HPLC. Blood glucose level was estimated spectrophotometrically using commercial glucose assay kit. In vitro release and pharmacokinetic profile of niosomal formulation and free insulin were evaluated. Though there was a slight delay in the in vitro drug release due to cholesterol content in the niosomes, there was no difference between the two preparations when plasma levels were compared in vivo. Niosomes significantly reduced the blood glucose level in diabetic rats. Fall in blood glucose level was almost 92% of initial value. In case of the niosomal form the half-life of insulin was prolonged by 4 -5 hr in contrast to 2 hr for free drug. Niosomes maintained the plasma insulin level up to 12 hr, but free drug was cleared quickly. The area under the plasma concentration-time curve for niosomal forms was, 26.07 degrees +/- 0.99 mIU. hr/ml and for free insulin was 11.722 +/- 1.00 mIU. hr/ml. More than 80% of the drug was successfully encapsulated to give a formulation with sustained release characteristics. Entrapment efficiency increased with increasing lipid concentration and decreased with increasing drug concentration. The results showed that insulin entrapped in niosomes prolongs the existence of drug in the body therefore increasing its therapeutic value. PMID- 12561950 TI - Effect of aspirin on blood-lipid interaction and lipid peroxidation phenomena in relation to partition coefficient and biological activity. AB - Considering the lipophilicity of aspirin (log P = -1.15), a significant contributor to its action mechanism, interaction of the drug with the whole lipids of goat blood have been investigated using phospholipid binding and lipid peroxidation phenomena as the parameters under investigation. The lipid content change along with the peroxidation induced by aspirin and its suppression with ascorbic acid had been quantitatively measured. Significant loss in phospholipid was observed after incubation of whole blood with aspirin in varying periods of time. This may be ascribed to binding affinity of aspirin with lipid constituents in blood, which may have potential role in its therapeutic effect. Lipid peroxidation induction potential of aspirin caused significant extent of peroxidation. Ascorbic acid, an antioxidant could significantly reduce aspirin induced lipid peroxidation. PMID- 12561951 TI - Hepatic and renal oxidative stress in acute toxicity of N-nitrosodiethylamine in rats. AB - Nitrosoamines such as N-nitrosodiethylamine (NDEA) produce oxidative stress due to generation of reactive oxygen species and may alter antioxidant defence system in the tissues. NDEA was administered ip as a single dose to rats in LD50 or in lower amounts and the animals were sacrificed after 0-48 hr of treatment. The results showed that lipid peroxidation in liver increased, however no significant increase in kidney LPO was observed after NDEA administration. Superoxide dismutase (SOD) and glutathione reductase (GSH-R) activity increased in liver, however, catalase (CAT) activity in liver was inhibited in NDEA treated rats. Kidney showed an increase in SOD activity after an initial decrease along with increase in GSH-R activity in NDEA treated rats. However, kidney CAT activity was not significantly altered in NDEA intoxicated rats. Serum transaminases, serum alkaline phosphatase blood urea nitrogen, serum creatinine and scrum proteins were elevated in NDEA treated rats. The results indicate NDEA-induced oxidative stress and alteration in antioxidant enzymes in liver and kidney to neutralise oxidative stress. PMID- 12561952 TI - Expression of heat shock protein 70 in freshwater prawn Macrobrachium malcolmsonii (H. Milne Edwards) following exposure to Hg and Cu. AB - Juveniles of freshwater prawn M. malcolmsonii were exposed to 1/6th concentration of LC50 of Hg and Cu for 48 hr. Sampling was done at 1 8, 12, 16, 24, 30, 36, 42 and 48 hr of exposure. Gill and hepatopancreas were dissected and subjected to one-dimensional electrophoresis. Western blotting was employed to determine the relative concentration of heat shock protein, hsp 70 (stress-70) in each sample. In the gill tissue of the prawn that had been exposed to Hg (0.024 mgHg 1(-1)), stress-70 was detected from the 1st hr till the 16th hr of exposure. But in the gills of Cu exposed prawn, synthesis of stress-70 appeared from the 1st hr till the 24th hr. Synthesis of hsp70 was not recorded after the 24th hr in the gills of exposed prawns. Synthesis of stress-70 was also found to be tissue-specific for both metals in this prawn. When the antibody probe raised against stress-70 was used, synthesis of stress-70 was not observed in hepatopancreas of prawns exposed to Hg or Cu, during the entire period of exposure of 48 hr. The expression of stress-70 in M. malcolmsonii following exposure to Hg and Cu is apparently only transient, and also a differential expression of stress-70 between gill and hepatopancreas was observed for both the metals. PMID- 12561953 TI - Auxin regulated poly(A)polymerase activity in Cicer arietinum epicotyls. AB - There was a linear increase in poly (A+) polymerase activity in the C. arietinum epicotyls during germination. Six-day-old auxin treated seedlings showed about 3 4 fold stimulation in enzyme activity, accompanied with 3- fold rise in the relative abundance of poly (A+) RNA levels. Actinomycin D, cycloheximide, cordycepin and amino acid analogues caused dramatic decline in poly (A+) polymerase as well as poly (A+) RNA levels. It seems that auxin induced a de novo synthesis of this enzyme. PMID- 12561954 TI - Characterization of carbamoyl phosphate synthetase of Streptomyces spp. AB - Carbamoyl phosphate synthetase (CPS) activity in Streptomyces lividans was repressed (70%) by addition of arginine and uracil in the growth medium. Enzyme activity was also inhibited by UMP and activated by ornithine and IMP. Pattern of inhibition and activation was similar irrespective of whether the cells were grown in medium supplemented with arginine or with uracil. A mutant of S. coelicolor with dual auxotrophy for arginine and uracil possessed only about 20% of CPS activity compared to the wild-type strain. An activity staining protocol has been developed for CPS enzyme. Using this method a single CPS band has been observed in the crude extracts of Escherichia coli as well as in S. lividans. Taken together, our results supported the conclusion that Streptomyces species might possess a single CPS enzyme unlike other gram-positive bacteria, which show the presence of two pathway-specific isozymes (Bacillus) or none (Lactobacillus and Leuconostoc). PMID- 12561955 TI - PTTH--a potential growth activator in silkworm, Bombyx mori L. for enhancing silk production. AB - In silkworm, prothoracicotropic hormone (PTTH), directly or indirectly controls silk production and spinning activity along with juvenile hormone (JH). An effort was made to exploit the potential of PTTH by indirectly activating silk gland for increasing silk productivity using short chain synthetic analogues of PTTH. The analogy in action was also established using PTTH extracted from the silkmoth. Different doses of 42 synthetic PTTH analogues, viz., 2.5, 5, 10 and 20ppm and 3.3 mg/ml of PTTH extracted from silkmoth heads were administered orally to V instar silkworm larvae (Race:KAxNB4D2 and PMxNB4D2) at 0-144 hr at an interval of 24 hr. The analysed data showed an improvement of about 14 - 23% in KA x NB4D2 and about 10-14% in PMxNB4D2 in respect of cocoon shell weight on administration of some of the synthetic PTTH analogues. The PTTH extracted from the adult brain also showed similar effect. The structural analogy of synthetic PTTHs (which improved the shell weight) with original PTTH and its probable mode of action in silkworm are discussed. PMID- 12561956 TI - Fertilising capacity of human sperms: a simple predictive assay. AB - Male subfertility is a growing reason for assisted reproduction. A limiting factor in male subfertility is asthenospermia. Motility is a cardinal indication of sperm vitality. Thus prognostic assays are aimed at quantitative determination of progression to assess the fertilising potential. However, a method permitting reliable prognosis of the fertilising capacity has yet to be developed. The assay presented here is the outcome of empirical data based on 590 IVF (in vitro fertilisation) trials. It is essentially a further exploitation of the Swim Up procedure, the selected sperms being maintained in culture under identical conditions employed in IVF. Semi-quantitative daily recordings of linear progression until complete extinction provided an index on vitality which is directly related to the fertilising potential. The findings indicated that a threshold of 50% linear motility after 24 hr culture was required to initiate fertilisation. The fertilising potential was guaranteed when at least 60% linear motility was observed at 24 hr, making the assay a predictive one. Its simplicity is an attractive feature. PMID- 12561957 TI - Effect of Emblica officinalis tannoids on a rat model of tardive dyskinesia. AB - Effect of active tannoid principles of E. officinalis, comprising of emblicanin A (37%), emblicanin B (33%), punigluconin (12%) and pedunculagin (14%), was investigated on a rat model of tardive dyskinesia (TD) induced by once daily administration of haloperidol (1.5 mg/kg, ip) for 28 days. Involuntary orofacial movements (chewing movements, buccal tremors and tongue protusion) were assessed as TD parameters. The tannoid principles of E. officinalis (EOT) were administered concomitantly with haloperidol in the doses of 10, 20 and 50 mg/kg, po, for 28 days. Sodium valproate (200 mg/kg, po), a Gaba-mimetic agent, and vitamin E (400 mg/kg, po), an antioxidant, were used as the standard drugs and administered for the same period. EOT induced a dose-related inhibition of all the three TD parameters assessed, as did vitamin E. The effect of sodium valproate remained statistically insignificant. The results suggest that EOT exerts a prophylactive effect against neuroleptic-induced TD which is likely to be due to its earlier reported antioxidant effects in rat brain areas, including striatum. PMID- 12561958 TI - Aluminium related changes in brain histology: protection by calcium and nifedipine. AB - Aluminium (Al; 50 mg AlCl3/kg body wt/day) treatment caused a marked change in histological picture of normal brain as indicated by an increased number of vacuolated spaces. These changes returned to normal partially by simultaneous treatment with nifedipine (0.7 mg/kg body wt/day) and completely by similar treatment with 50 ppm calcium (CaCl2; 12.5 mg/kg body wt./day). Neither nifedipine nor calcium treatment alone altered the normal histological condition. The histological changes could not be correlated with the decrease in calcineurin activities in brain as nifedipine decreases calcineurin activity without any histological changes. Hence the histological changes may be considered as specific for Al and not due to a general decrease in calcineurin activity. PMID- 12561959 TI - RAPD markers linked to brown planthopper Nilaparvatha lugens resistance locus in rice. AB - Brown plant hopper, a major pest in rice causes "hopper burn" in the field. The resistance gene for brown planthopper was mapped by using 20 recombinant inbred lines (RIL's) derived from a cross between resistant line Oryza. officinalis derivative (IR 54742-2-21-12-17-6) and a susceptible rice cultivar ASD 16 using bulked segregant analysis. On an average of 4 loci were amplified and two RAPD primers amplified loci that co-segregated with resistance/susceptibility. The segregating RAPD loci were mapped using Mapmaker programme into 13 groups. The expected and the 95% confidence level were found to be 15.2 and 47.7 cM respectively, confirming the location of the brown planthopper resistant gene on the region of chromosome 4. These RAPD markers will accelerate breeding programme for brown planthopper resistance. PMID- 12561960 TI - Susceptibility of diamond back moth, Plutella xylostella (L) to entomopathogenic nematodes. AB - Eight entomopathogenic nematode species / strains, Steinernema glaseri (steiner), S. carpocapsae (Weiser), S. feltiae (Filipjev), Steinernema sp. Ecomax strain, Heterorhabditis bacteriophora (Pioner), Heterorhabditis sp. Ecomax strain, two locally isolated strains called as JFC and TFC were tested against the final instar larvae of diamond back moth, Plutella xylostella (L.). All nematodes were found pathogenic. However, H. bacteriophora was adjudged the most pathogenic amongst the test nematodes on the basis of LD50 (9.16 IJS/larva), LT50 (43.26 hr), Lex T50 (3.24 hr) and the propagation potential (average of 271.42 IJS/mg) on the host body weight. PMID- 12561961 TI - Herbal formulations as pharmacotherapeutic agents. AB - Herbal medicines are popular as remedies for diseases by vast majority of world's population. Polyherbal preparations are products from medicinal plants. These are considered as safe since they are natural products. Herbal formulations which have reached widespread acceptability as therapeutic agents in India include noortropics, antidiabetics, hepatoprotective agents and lipid lowering agents. Pharmacological effects of many plants have been studied in various laboratories in India. However, there are many limitations regarding safety and efficacy of these preparations. Knowledge about active principles of herbal preparations are not well defined, information on toxicity and adverse effects of these formulations are lacking. Information regarding pharmacokinetics and bioavailability is not available. Packet inserts providing details regarding safety, and warning are not required for sale of these which are available as over the counter preparations. The risk of untested and unregulated remedies should be known to the lay public. Selection of plant material based on quality, standardisation of methods of preparation, enforcement of regulation regarding appropriate labels, are measures which will improve the quality and acceptability of herbal preparations as therapeutic agents. Documentation of research publications in journals and availability of information on website, are other measures to assist research in this field. PMID- 12561962 TI - Low level radiation exposure the radiobiologist's challenge in the next millennium. AB - A formal definition for low level exposure does not exist. This has arbitrarily been defined here as exposures from 0 to 5 cGy. The health implications of exposures within this dose range are highly controversial since the effects are exclusively stochastic. As such, the effects can only be detected in large populations. The Oxford Survey of Childhood Cancers (OSCC) established leukaemia as a predominant effect. After the chernobyl nuclear disaster, studies in European countries have correlated perinatal mortality with radioactive contaminations which could only have raised the radiation burdens by levels which are currently regarded as negligible. The reported risk indices for childhood leukaemia arising from low level exposures are generally comparable to those ascertained for high exposures, thus posing an enigma to radiobiologists. This paper reviews the progress in various areas of radiobiological research and attempts to make a synthesis of the facts with the view to provide an explanation. The purpose is also to stimulate an understanding of multifactorial biological mechanisms. Environmental radiation exposures must be expected to be concomitant with other toxic agents which must be taken into account in risk assessment. The challenge in the future will be to realise this goal. PMID- 12561963 TI - Insulin like growth factor 1 and regulation of ovarian function in mammals. AB - Various growth factors have been proposed to play endocrine and/or paracrine role in mammalian ovarian follicular development. The insulin like growth factor 1 (IGF-1) is one such factor. More and more reports now support the existence of an intra-ovarian IGF system including receptors and binding proteins. The role of IGF-1 in ovary is to amplify gonadotropin hormone action in terms of increased steroidogenesis by ovarian granulosa cell and granulosa cell proliferation. The synthesis and proteolysis of insulin like growth factor binding proteins, under the control of follicle stimulating hormone, regulate the intra-follicular availability of IGF-1, which further determines the sensitivity of granulosa cells to gonadotropins. Besides gonadotropins, IGF-1 has been implicated in somatotropin hormone action in the ovarian function. Exact mechanism of IGF-1 action in the ovarian follicles needs to be worked out to elucidate whether or not IGF-1 is indispensable in addition to know endocrine factors like gonadotropic and ovarian steroid hormones. This will pave the way for better understanding of control(s) which ensure final development of dominant follicle(s) and atresia of other follicles of the cohort. PMID- 12561964 TI - Effect of chronic treatment with losartan on streptozotocin induced diabetic rats. AB - Treatment of rats with streptozotocin (STZ, 45mg/kg, i.v.,single dose) produced cardinal symptoms of diabetes mellitus including hyperglycemia, hypoinsulinemia and increase in blood pressure. Treatment with losartan--an angiotensin (AT1) receptor antagonist, 2 mg/kg, po for 6 weeks decreased the blood glucose levels by 16.5%. There was 190% increase in AUCglucose and 59.4% decrease in AUCinsulin in STZ-diabetic rats as compared to control rats. Treatment with losartan caused slight decrease in AUCglucose and slight increase in AUCinsulin. There was no significant difference in insulin sensitivity (K(ITT)) index of STZ-diabetic group as compared to control. Losartan treatment failed to alter these levels significantly. Serum cholesterol and creatinine levels were found to be increased significantly in STZ-diabetic rats. Treatment with losartan significantly prevented the rise in cholesterol and creatinine levels by 20.1 and 81% respectively. The results suggest that losartan produces some beneficial effects in STZ-diabetic rats. PMID- 12561965 TI - Effect of prenatal alprazolam exposure on anxiety patterns in rat offspring. AB - Prenatal alprazolam (APZ) treatment in 0.1 and 0.2 mg/kg/day doses during 13-20 days of gestation induced significant increase in open-field ambulation, rearings, self-grooming and faecal pellets in rat offspring. Prenatal APZ treated rats displayed significantly increased anxiogenic behaviour on elevated plus maze (spent less time on open arms, more time on enclosed arms and made less number of entries on open arms) and increased anxiogenecity on elevated zero maz e(APZ treated rats spent less time on open arms and made less number of head dips and stretched attend postures in comparison to control rat offspring). The results indicate persistent behavioural alterations in the rat offspring after prenatal exposure to APZ. PMID- 12561966 TI - Male reproductive toxic effects of carbendazim: hitherto unreported targets in testis. AB - Carbendazim (MBC), a widely used fungicide, is toxic to male reproductive mechanisms. Various cellular targets in the testis for MBC action are being understood only recently and still more targets have been conceived. The present study was aimed at finding such newer targets. Male rats were administered through oral route a single dose of carbendazim (400 mg/kg) and the testis was studied adopting routine histological technique. It has been observed that pachytene spermatocytes could also be targets for MBC action in the testis. The study also reports selective loss of step 14 spermatids, asynchrony of the stages in the spermatogenic cycle and development of Sertoli cell fibrosis of the seminiferous tubules of carbendazim-treated rats. From the different kinds of responses seen in the seminiferous tubules in the same testis to MBC, particularly in the Sertoli cell, MBC action in the testis appears dependent on the stage in the spermatogenic cycle at first exposure. PMID- 12561967 TI - Effect of Ruta graveolens L. and Euphorbia peplus L. anti-inflammatory extracts on nutritional status of rats and the safety of their use. AB - A significant increase in body weight with remarkable increase in total food intake and significant increase in protein efficiency ratio were observed following oral administration of R. graveolens ether extract (500 mg/kg body wt) to growing rats for 3 weeks. Serum albumin was significantly decreased after administration of declofenac (15 mg/kg body wt). Albumin/globulin ratio decreased significantly on administration of E. peplus ether extract (500 mg/kg body wt). No significant changes were observed in other biochemical and nutritional parameters on administration of either of the extracts or declofenac. However, only a significant elevation of alkaline phosphatase was noticed during treatment with R. graveolens. The results suggest that both plant extracts have no harmful effect on nutritional status and are safe towards kidney functions, while Euphorbia is more safe than Ruta in relation to liver functions. PMID- 12561968 TI - Nutritional evaluation of extruded faba bean (Vicia faba L.) as a protein supplement in cereals based diet in rats. AB - The quality of extruded faba bean (EFB) (Vicia faba L.) as a protein supplement in cereals diet was evaluated. Growth rate and protein efficiency ratio was better in 20% EFB blended dietary group of rats than those fed either on cereals alone or at 40% EFB supplementation. Subsequent studies on digestibility co efficient, biological value, net protein utilization and net protein ratio also confirmed biological utilization of EFB proteins in a better way at low level (20%). PMID- 12561969 TI - Role of male scent glands in improving poison bait acceptance in female rats, Rattus norvegicus. AB - The poison (Zn3P2) bait mixed with preputial gland and cheek gland extract was highly effective as compared to poison bait mixed without the scent gland extract, in increasing the food consumption and mortality rate in rats. Among these two scent glands, preputial gland extract was more effective than cheek gland extract in increasing the bait acceptance. The scent gland extract mixed with poison bait was capable of improving the poison bait acceptance. A 10% concentration of scent gland extract was more effective than 5% concentration in acceptance of poison bait. The results suggest efficacy of scent gland in improving the poison bait acceptance. PMID- 12561970 TI - Prevention of HCl-ethanol induced gastric mucosal injury in rats by Garcinia cambogia extract and its possible mechanism of action. AB - Oral pretreatment of rats with G. cambogia fruit extract (1 g/kg body weight/day at interval of 7 and 15 days) protected gastric mucosa against HCl-ethanol induced damage by decreasing the volume and acidity of gastric juice. Increased lipid peroxidation, decreased activity of antioxidant enzymes, altered levels of protein and glycoproteins in the ulcerated mucosa, and gastric juice were maintained at near normal levels in G. cambogia pretreated rats. The results suggest the anti-ulcer activity of G. cambogia by virtue of its ability to decrease acidity and increase mucosal defense. PMID- 12561971 TI - Baculovirus studies in new, indigenous lepidopteran cell lines. AB - Eight lepidopteran cell lines were established recently and their susceptibility to different insect viruses was studied. Two Spodoptera litura cell lines from the larval and pupal ovaries, were found highly susceptible to S. litura nuclear polyhedrosis virus (SLNPV, 5-6 x 10(6) NPV/ml). The Helicoverpa armigera cell line from the embryonic tissue was highly susceptible to H. armigera NPV (HaNPV, 6.3 x 10(6) NPV/ml). These in vitro grown SLNPV and HaNPV caused 100% mortality to respective 2nd instar larvae. The susceptibility of the cryo-preserved cell lines to respective baculoviruses (SLNPV/HaNPV) was studied and no significant difference in their susceptibility status was observed. The cultures could grow as suspension culture on shakers and may find application for in vitro production of wild type/recombinant baculoviruses as bio-insecticides. S. litura and Bombyx mori cell lines from larval ovaries, were highly susceptible to Autographa californica NPV (5.5 x 10(6) NPV/ml) and Bombyx mori NPV (BmNPV, 6.1 x 10(6) NPV/ml) respectively. These cell lines may find application in baculovirus expression vector studies for the production of recombinant proteins, useful in the development of diagnostic kits or as vaccines. PMID- 12561973 TI - Responses of isolated scale melanophores of a fresh water carp, Cirrhinus mrigala (Ham.) to zidovudine. AB - Zidovudine (AZT) induced concentration related aggregation in C. mrigala melanophores. Denervated melanophores failed to respond to AZT. Specific and nonspecific alpha adrenoceptor antagonists completely blocked the responses of fish melanophores to AZT. Histamine and prostaglandin antagonists also inhibited aggregation of the melanophores induced by AZT. The results suggest that AZT may release a mixture of neurotransmitter like substances, which cause the aggregation of this fish melanophores. PMID- 12561972 TI - New Helicoverpa armigera Hbn cell line from larval hemocyte for baculovirus studies. AB - A new cell line from the larval hemocytes of H. armigera was established in Grace's medium modified by adding lactalbumin hydrolysate and yeastolate (3.3g/l), and supplemented with fetal bovine serum (20%). The cell line was designated as NIV-HA-1195. The cell population at P-78 consisted mainly of epithelial-like cells (89.36%), fibroblast-like cells (8.31%) and giant cells (2.13%). The population doubling time was 96hr at P-8, 60hr at P-43. The chromosome number ranged from 45 to 200. The cell line is susceptible to the baculoviruses, Autographa californica nucleopolyhedrovirus (AcNPV), Spodoptera litura NPV and the homologous HaNPV. Isoenzyme profile and results of 16S rRNA heteroduplex analysis clearly indicated the species specificity of the new cell line. PMID- 12561974 TI - Evidence for presence of GABA-ergic receptor mediated dispersion in isolated scale melanophores of a carp, Cirrhinus mrigala Ham. AB - Effects of GABA-ergic agonists and antagonists were examined on the melanophores of a carp C. mrigala in vitro. GABA and baclofen both induced concentration - related dispersion in fish melanophores. Denervation of the melanophores by reserpine treatment potentiated the sensitivity of the melanophores to GABA. While denervation by cooling treatment inhibited the sensitivity of the melanophores to GABA, atropine, bicuculline and pentylenetetrazole all inhibited the dispersal responses of the melanophores induced by higher concentrations of GABA. 5-aminovaleric acid also significantly inhibited the dispersion of the melanophores induced either by GABA or baclofen. It is concluded that GABA-ergic agonist induced dispersal responses in C mrigala melanophores are mediated through specific GABA receptors. The presence of both GABAA and GABAB receptors in this fish melanophores has been indicated. PMID- 12561975 TI - Effect of formulated diet on digestive enzymes of Labeo rohita (Ham.). AB - Six sets of feeding experiments were carried out using formulated diets containing prawn head waste (PW), chicken intestine waste (CW), banana flower (BF), cauliflower waste (CAU) Dolicos lab lab (DLL) and groundnut leaf (GNL) in four levels of inclusion (15, 30, 45 and 60%) to assess the pattern of distribution and activities of digestive enzymes like cellulase, amylase, maltase, invertase, protease and lipase in the digestive tracts of Labeo rohita fingerlings. A control group of fish was fed with diets containing antibiotics to destroy the digestive tract microflora which may induce digestive functions. In general, the activity of digestive enzymes depended on the amount and type of the ingredients present in the diets ingested by the fish. Test animals showed both endogenous and bacterial cellulase activities which suggests the necessity for including cellulose (plant protein source) as dietary ingredient. Occurrence of higher amount of cellulase in the foregut and amylase in the fore and midgut influenced by DNL and GNL diets revealed the possibility of including less than 40% of the respective ingredients in the diet of rohu. Maltase and invertase were highly influenced by GNL, DLL and BF diets than PW and CW diets. More than 40% inclusion of PW and CW was found to increase protease and lipase secretion in the midgut and hindgut regions. The higher secretion of lipase in the midgut suggested the physiological versatility for lipid digestion in rohu fingerlings. PMID- 12561976 TI - Effect of alternaria pathotoxin(s) on expression of p53-like apoptotic protein in calli and leaves of Brassica campestris. AB - Possible involvement of apoptosis was investigated in pathotoxin-treated and nutritionally-depleted in vitro cultured calli by comparing levels of p53-like protein. Antibodies raised against human p53 were used to detect and quantify p53 in B. campestris. Expression of p53-like protein increased from proliferating to static growth stage and reached to constant level at decaying stage. Both ELISA and dot immuno-binding assay showed that p53-like protein was over expressed in toxin treated and nutritionally depleted calli. Almost similar changes were seen in senescent damage in Brassica species indicating involvement of p53 dependent pathways. PMID- 12561977 TI - Environmental radiation as the conditioning factor for the survival of yeast Saccharomyces cerevisiae. AB - Whether natural radiation can be a conditioning factor for the growth and survival of a living organism was investigated using diploid yeast S. cerevisiae D7. Yeast cells were conditioned by growing them continuously for at least 100 generation in 3 different radiation background such as i) ambient radiation (1.1 mSv/y), ii) sub-ambient radiation (0.44 mSv/y, within a shielded chamber) and iii) an elevated background radiation (88 and 880 mSv/y in a gamma-field). At the end, the cells were challenged with 60Co gamma, 100 Gy and the viable fractions were determined. Conditioning the cells in 880 mSv/y and in ambient radiation, enabled the cells to reduce the deleterious effect of the challenging dose significantly (P < 0.05) compared to that of sub-ambient radiation. The cellular viability of yeast cultures seems to be influenced by the prevailing radiation background, apart from starvation. Comparatively, a rapid decline in viability was noticed when the cultures were incubated for 60 days in the shielded chamber. The results indicate that some amount of radiation equivalent to background level or little above is needed to confer fitness in biological systems against stress factors, including radiation. The adaptive dose for the diploid yeast was also determined by single exposure. The priming dose ranged from 0.01 to 1.2 Gy. An adaptive dose of 0.25 or 0.4 Gy, almost nullified the deleterious effect of the challenging dose. The adaptive response may have a greater role in the field of cancer therapy and in radiation risk assessment. Understanding the response of an organism at different radiation-background will be helpful for successful space management. PMID- 12561978 TI - Degradation of predigested distillery effluent by isolated bacterial strains. AB - Batch studies were conducted on degradation of anaerobically digested distillery wastewater by three bacterial strains, viz. Xanthomonas fragariae, Bacillus megaterium and Bacillus cereus in free and immobilized form, isolated from the activated sludge of a distillery wastewater treatment plant. The removal of COD and colour with all the three strains increased with time up to 48 hr and only marginal increase in COD and colour removal efficiency was observed beyond this period up to 72 hr. After this period removal efficiency remained fairly constant up to 120 hr. The maximum COD and colour removal efficiency varied from 66 to 81% and 65 to 75%, respectively for both free and immobilized cells of all the three strains. The strain Bacillus cereus showed the maximum efficiency of COD (81%) and colour (75%) removal out of the three strains. An interrelationship between the percent COD and colour removal was carried out by correlation and regression analysis and was justified by high values of coefficient of correlation (r = 0.99) for all the cases. The first order removal rate kinetics was also applied and rate constants were evaluated for COD and colour removal efficiencies. PMID- 12561979 TI - Heat modifiability of outer membrane protein of Pasteurella multocida serotype B:2. AB - Outer membrane proteins (OMP) are generally porins, functioning as molecular sieves assisting in the transmembrane transportation. Heat modifiable characteristics of OMP from P. multocida B: 2 have been explored to know their basic characteristics on event of temperature rise. A major band of 32 kDa and two minor bands of approximately 39 and approximately 28 kDa were found to be heat modifiable. It is suggested that boiling at 100 degrees C in presence of beta mercaptoethanol for 5 min is sufficient for characterisation of OMP by Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis. PMID- 12561980 TI - Cloning and sequencing of beta toxin gene of Clostridium perfringens type C. AB - A gene encoding beta toxin was amplified by polymerase chain reaction from C. perfringens type C isolate and cloned in pUC 19 vector. The nucleotide sequence was identical with C. perfringens type B beta toxin gene sequence. The Southern hybridization using labelled beta toxin gene probe revealed the presence of positive signals only in beta producing C. perfingens. PMID- 12561981 TI - Effect of ciprofloxacin on specific immune response in rabbits. AB - Ciprofloxacin (10 mg/kg body weight, iv, twice daily for 4 days) failed to alter specific antibody titres, total immunoglobulin concentration, total serum protein concentration, total leukocyte count, lymphocyte percentage, phagocytic index and skin thickness in DNCB skin sensitivity test against Brucella plain killed antigen in New Zealand White rabbits. It can be concluded that ciprofloxacin at the dose and duration employed did not adversely affect specific immune response in normal rabbits. PMID- 12561982 TI - Pharmacological and haematological study of shol fish (Channa striatus) skin extract on experimental animal. AB - Snake head fish Channa striatus (locally called 'shol') skin extract (SFSE) was examined for certain pharmacological and haematological effects on experimental animals. LD50 of SFSE was found to be 6 mg/20gm (iv) in male albino mice. SFSE potentiated pentobarbitone induced sleeping time in male albino mice and produced hypothermia. Low dose of SFSE decreased respiratory rate in rat and guineapig and high dose produced apnoea leading to death. On isolated toad and guineapig heart, SFSE significantly decreased rate and amplitude of contraction leading to temporary blockade, which returned after repeated wash. On isolated nerve muscle preparations, SFSE produced irreversible blockade of twitch response. SFSE induced quick contraction on isolated guineapig ileum, which was antagonised by atropine and cyproheptadine. SFSE did not possess haemolytic and haemorrhagic activity but produced anaemia in male albino mice. A neurotoxic compound (fluoroscent and ninhydrin positive) was isolated from SFSE by thin layer chromatography. This compound (CS-NT) was lethal in male albino mice, produced death by apnoea in rat and produced irreversible blockade of isolated nerve muscle preparation. This study confirms that the skin of Channa striatus possesses toxic, and lethal components, which needs further detailed study. PMID- 12561983 TI - Use of meristem tip culture to eliminate carnation latent virus from carnation plants. AB - A successful protocol for meristem tip culture to eliminate carnation latent virus from carnation cv. scania has been described . The virus was found to be mechanically transmissible to Chenopodium quinoa, C. amaranticolor, Dianthus barbatus and Saponaria vaccaria. Murashige and Skoog'smedium (MS) supplemented with NAA (1.0 microM) and Kn (20.0 microM) proved best for meristem establishment and microshoots were rooted in MS medium supplemented with IBA (5.0 microM). Meristems measuring 0.1 and-0.2 mm yielded virus free plants and larger meristems were not effective. PMID- 12561984 TI - 49th I.A.P.M. Conference APCON-2000, at Safdarjung Hospital, New Delhi. PMID- 12561985 TI - AIDS related malignant disease at regional institute of medical sciences. AB - 26 AIDS patients within a period of 3 years were studied for associated malignant disease. There were 4 associated malignancy. The commonest AIDS associated malignant disease found is Kaposi's sarcoma seen in 2 patients. In one each of the AIDS patients, association of Non-Hodgkin's lymphoma and acute Promyelocytic leukemia (M3) was found. Findings by other workers are reviewed. With new trends in management of AIDS patients that prolong their survival, an increase in the incidence of AIDS associated malignancy is anticipated. PMID- 12561986 TI - Cardiac pathology of permanent pacemaker implementation. AB - The cardiac findings in five cases of permanent pacemaker implantation seen at autopsy in a three-year period is reported. Implantation was done for complete heart block in four patients and sick sinus syndrome in one. The periods of implantation ranged from seven days to four years. The common findings were right sided valvar and mural bland thrombus formation. Additional findings included superior vena caval thrombosis, endocardial 'tunnel' formation for the pacing wires, perforation of the tricuspid leaflet and a gross morphology similar to that seen in right sided endomyocardial fibrosis. PMID- 12561987 TI - A study of the haematologic spectrum of myelodysplastic syndrome. AB - In 31 patients of myelodysplastic syndrome, RAEB-t was the commonest subtype (29%), and RARS, the lease common (6.4%); 19.4% were characterised as the unclassifiable (UC) group. Pallor was the dominant sign (90.3%). Low haemoglobin in RA & RARS (p<0.05), thrombocytopenia in RAEB-t (p<0.01) and high leuco/monocyte counts in CMML (p<0.001) were observed. Neutropenia occurred most frequently in RAEB & RAEB-t and circulating blasts in all cases of RAEB-t and CMML. Bicytopenia was the commonest finding (58.1%) and pancytopenia the least (16.1%). 84% of marrows were hypercellular and trilineage dysplasia was seen in 68% of patients. Megaloblastoid dyserythropoiesis was the predominant feature in all cases, dysgranulopoiesis in all cases of RAEB, RAEB-t and CMML, and micromegokaryocytes in all cases of RARS, RAEB & CMML were seen. RAEB-t and RAEB (33.3% each) were the predominant groups which progressed to leukemia, FAB AML M2, being the commonest type (60%). PMID- 12561988 TI - Imagecytometry: a new tool for diagnosis of glomerular haematuria. AB - Differentiation between glomerular and nonglomerular haematuria is a major challenge in clinical medicine, which is very important for a definitive diagnosis and management in individual cases. Phase contrast microscopy of red cells in urine is the standard practice for diagnosis of glomerular haematuria. Urine cell flowcytometry is recently being used for such diagnosis. In this context, the role of determination of haemoglobin content of urine red cells is not know. Application of image analysis to study the red cells in urine may be more objective and accurate for the diagnosis. The present study has been undertaken to evaluate the urine red cells with the help of an automated computerized image analysis system for determination of hemoglobin content by integrated optical density (IOD). The morphometric parameters were also analyzed. The glomerular RBCs were significantly smaller in diameter, area and perimeter than nonglomerular RBCs with a greater variation in shape and lower [OD (p<0.0001 to <0.00002). With the help of morphometric parameters the percentage of cases diagnosed correctly varied from 90 to 95. The IOD helped to diagnose 100% cases. Thus application of this new technique may be very useful diagnostic tool in the investigation of haematuria. PMID- 12561989 TI - Rhinosporidiosis--a clinicopathological study of 34 cases. AB - Rhinosporidiosis was the commonest (68%) fungal lesion encountered during the period of 11 1/2 years from January 1987 to July 1998. Men in 2nd, 3rd, 4th decade were commonly affected. Nose and nasopharynx were the commonest (85%) sites involved followed by ocular tissue (9%). One patient had involvement of bone (tibia). Generally a lymphoplasmacytic response was observed in all cases. Polymorphonuclear leukocytic response mostly observed at the site of rupture of sporangia. Epithelioid cell granulomatous and giant cell response observed in 47% of cases. Transepithelial migration of sporangia observed in 76% of cases. Rhinosporidium seeberi could be easily identified in haematoxylin and eosin stained sections. The walls of young trophic forms are delineated well with the PAS stain and verhoeff van Gieson stain. PMID- 12561990 TI - Glucose-6-phosphate dehydrogenase deficiency in blood donors: screening by micromethaemoglobin reduction test. AB - 500 blood donors were screened for G6PD deeiciency using micromethaemoglobin reduction (microMRT) test. Most of the blood donors were young adult males (95.8%). The overall incidence of G6PD deficiency was found to be 0.8%. There, was apparently decreased frequency of G6PD deficient blood donors with increasing age, and no relation could be ascertained between G6PD) deficiency and blood groups. PMID- 12561991 TI - Pattern of cancer in Dayanand Medical College & Hospital, Ludhiana (a ten year retrospective study). AB - A ten year retrospective study was undertaken to ascertain the pattern and incidence of cancer in Ludhiana which is an industrial town with catchment area of Ludhiana consisting of Ludhiana distt., Sangrur, Jalandhar, Hoshiarpur, Faridkot, Ferozepur, Ropar, Kapurthala, some parts of Himachel Pradesh and Haryana. A total number of 56,565 biopsies were received for histopathological examinaiton in 10 years and 4,730 cases of cancer were diagnosed. The incidence of total malignant tumors was 8.36%. Females out numbered males in the incidence of cancer, with male to female ratio being 1:1.09. Most of the cancers were seen in the age group of 41-50 years. In females the two most common cancer sites were breast (21.07%) and cervix (19.4%) while in males hypopharynx--larynx (13.94%) and prostate (9.65%) were the most common sites of cancer. PMID- 12561992 TI - Factors regulating the metastatic potential of benign giant cell tumour of bone- study of an unusual case with short review of literature. AB - Benign giant cell tumour of bone with metastases to other bones and lungs is extremely rare. Benign metastasising giant cell tumour is distinctly separate from multicentric giant cell tumour, primary and secondary malignant giant cell tumour. Factors regulating the local recurrence and metastatic potential of this benign tumour depend on its aggressiveness which can be better assessed by clinical and radiological parameters rather than the histopathological appearance. A benign giant cell tumour of ischium with metastasis to vertebra and lung over an eleven year period is discussed. Extreme paucity of literature prompted to publish the article. A short review of factors determining the recurrence and metastatic spread of benign giant cell tumour of bone is highlighted. PMID- 12561993 TI - Vaginal carriage of group B Streptococcus in infertile women. AB - From May, 1999 to April, 2000, 256 high vaginal swabs were culture from asymptomatic infertile women attending the out-patient department of the Institute of Reproductive Medicine. 41 strains of Streptococcus agalactiae were isolated from 33 patients (12.89%). Five patients had repeated isolations even after adequate therapy. PMID- 12561994 TI - Experience of human rabies in north India. AB - Rabies is one of the most common causes of human encephalitis in developing countries. This study shows the diagnosis of rabies among suspected human rabies encephalitis cases by Seller stain, Flourescent stain as well as mouse inoculation test. Out of 71 postmortem brain specimens, 26 were diagnosed as rabies positive. Negri bodies were demonstrated in 18 (25.4%) brain saples by Seller stain. Flourescent antibody technique could detect rabies antigen in 21 (29.6%) samples. Rabies virus could be isolated in 15 (42.9%) of the 35 samples by intracerebral inoculation in 15 (42.9%) of the 35 samples of intracerebral inoculation in newborn Swiss albino mice. Of the 26 confirmed cases, 61.5% occurred during the months of June to August and history of dog bite was present in 9 (34.6%) cases. PMID- 12561995 TI - Candidemia in neonatal intensive care unit. AB - The present study was conducted over a period of 6 months to determine the Candida species causing candidemia in a neonatal intensive care unit and to analyse the risk factors associated with acquisition of significant fungemia. Speciation of the 19 isolated Candida spp was done by the standard techniques. Antimicrobial susceptibility of these isolates was determined by disc diffusion method against Amphotericin B, Fluconazole, Ketoconozole and 5-Flucytosine. Candida glabrata was the most common species involved (42.1%). Other species isolated were C. tropicalis (31.6%). Calbicans (21.1%) and C.parapsilosis (5.2%). All the isolates were sensitive to Amphotericin B. Resistance to other antigungal agents was seen only in C. globrata. Significant candidemia was seen in 14/19 (72.6%) of neonates. Risk factors found to be associated with significant candidemis in these neonates included intake of multiple broad-spectrum antibiotics (p<0.0001), use of total parenteral nutrition (p<0.045) and ventilators (p<0.0001). PMID- 12561996 TI - Smears from rat liver suspension as modified substrate for antinuclear antibody test. AB - Immunofluorescence still remains a standard method for documenting antinuclear antibodies (ANA). Cryostat cut sections of rodent liver or Hep2 cell nuclei have been used as substrate in the test but are often difficult to arrange in laboratories in developing countries. Hence, a modification was developed using smears from rat liver suspensions. The smears were compared with the cryostat cut sections over 338 sera samples of suspected cases of collagen diseases, rheumatoid arthritis, thyroid disorders, hepatitis B, enteric fever, tuberculosis and normal subjects. The sera from suspected collagen diseases cases were also compared with ANA test using Hep2 cells. The modified smear technique was well comparable and the clarity of the immunofluorescence was even better than for cryostat cut sections. Using the modified smear technique 272 sera out of 2,851 sera gave positive test for ANA. The homogenous, speckled and peripheral patterns were seen for 203, 66 and 3 samples respectively. To conclude: The smears prepared from homogenised rat liver suspension and fixed like bacterial smears offer a very convenient and reliable tissue substrate for ANA test. PMID- 12561997 TI - Gelatinous transformation of bone marrow following chemotherapy for myeloma. AB - Gelatinous transformation of the marrow (GTBM) has been associated with various conditions. We present a unique case of GTBM in a patient with myeloma following treatment with Melphalan. PMID- 12561998 TI - Endometrial stromal sarcoma with sex-cord like features. AB - Endometrial stromal sarcomas are rare, low grade, malignant uterine tumours. Sometimes, they manifest an epithelial like or sex-cord like differentiation. This is a report of one such case in a 35 year old female. PMID- 12561999 TI - Malignant fibrous histocytoma of bone in neurofibromatosis--a case report. AB - Multiorgan neoplasms are encountered frequently in patients with neurofibromatosis, which is a genetic disorder. Though skeletal abnormalities are common in these patients, the occurrence of primary bone sarcomas is very rare. We hereby report a case of post-radiation malignant fibrous-histiocytoma of bone developing in an elderly patient with neurofibromatosis. PMID- 12562000 TI - An incidental case of breast cysticercosis associated with fibroadenoma. AB - It is unusual for cysticercosis to occur in the substance of the breast. Only a few cases have been described in the literature. We report a case of cysticercosis perched on fibroadenoma of breast found on surgical excision in a 35 year old Nepali woman. The patient had no discernible evidence of cysticercosis in the other organs. PMID- 12562001 TI - Malignant Sertoli cell tumour--a case report. AB - Sertoli cell tumours are rare sexcord stromal tumours of testis. Malignant behaviour is observed in one tenth of such tumours. A malignant sertoli cell tumour is reported here in a 70 years old man. The tumour was of large size and showed necrosis, marked celllar pleomorphism, and mitotic figures. PMID- 12562003 TI - Niemann Pick disease--a case report. AB - Niemann Pick disease is a rare disorder occuring in paediatric age group. The clinical features, hematological characteristics and autopsy findings in an one year old male diagnosed to have Niemann Pick disease are presented. PMID- 12562002 TI - Botryoid rhabdomyosarcoma of prostate--a case report. AB - Botryoid rhabdomyosarcoma of the prostate is a rare tumour in children, mostly below 6 years of age. A case of 5 year old male child presenting with acute retention of urine due to botryoid rhabdomyosarcoma of the prostate is described. PMID- 12562004 TI - Piloleiomyoma--a report of five cases. AB - Piloleiomyomas are uncommon smooth muscle neoplasms of the skin with a few reproted cases in Indian literature (1,2,3,4,5). They are often misdiagnosed clinically. A correct biopsy report is important because patients may have to be managed medically since surgery is associated with a high rate of recurrence. The classical histologic findings, and Masson's stain to confirm the smooth muscle origin aids in the correct diagnosis. PMID- 12562005 TI - Haematological findings in neonates with sepsis. PMID- 12562006 TI - Supravital staining and bright field microscopy: a simple technique for urine sediment analysis. PMID- 12562007 TI - Acute myeloid leukemia (M4) presenting as generalized urticaria and papulonodular lesions on skin. PMID- 12562008 TI - Mycetoma at unusual site--orbit & patella. PMID- 12562009 TI - Prevalence of acinetobacter anitratus in urinary isolates at G.M. & Associated Hospitals, KC's Medical College, Lucknow, UP. PMID- 12562010 TI - Internet resources for the human geneticist. AB - The Internet is a massive expanding body of information, which is likely to play a significant role for clinicians and researchers across the world. Since its inception in December 1969 the Internet has grown rapidly and is anticipated to expand 1,000% in the coming next few years. Various useful databases on human genetics are already in 'the Net' and many more are being added constantly. The future of human geneticist is in handling of information. In this review of Internet and compilation of important web site addresses we expect to stimulate and instruct human geneticists in navigating the Net. The list of web sites provided in this article is expected to facilitate their search. PMID- 12562011 TI - Pharmacological activities of Genistein, an isoflavone from soy (Glycine max): part I--anti-cancer activity. AB - Phytoestrogens represent a family of plant compounds such as isoflavones, flavones and lignans. A variety of these plant compounds have been identified in various human body fluids. A wide range of commonly consumed foods contains appreciable amounts of these different phytoestrogens, viz. soy products are particularly good sources of isoflavones and lignans. Accumulating evidences from molecular and cellular biology experiments, animal studies, and to a limited extent, human clinical trials suggests that phytoestrogens may potentially confer health benefits related to various cancers and diseases such as cardiovascular disorder. The evidences reviewed here represent the beneficial effects of most potential and promising isoflavone, Genistein in various types of cancers. PMID- 12562012 TI - Pharmacological activities of Genistein, an isoflavone from soy (Glycine max): part II--anti-cholesterol activity, effects on osteoporosis & menopausal symptoms. AB - Phytoestrogens represent a family of plant compounds such as isoflavones, flavones and lignans. A wide range of commonly consumed foods contains appreciable amounts of different phytoestrogens such as isoflavones and lignans. Soy and its products are particularly good sources of isoflavones and lignans. The evidence reviewed here represents the beneficial effects of most potential and promising isoflavone, Genistein in various types of diseases such as osteoporosis, cardiovascular diseases, menopausal symptoms by accumulating evidence from molecular and cellular biology experiments, animal studies, and, to a limited extent, human clinical trials. This review suggests that phytoestrogens may potentially confer health benefits related to various diseases such as cardiovascular disorder, menopausal symptoms, and osteoporosis. PMID- 12562013 TI - Radiolabelled monoclonal antibodies against human cardiac myosin. AB - Myocardial infarction (MI) can be monitored using several protein markers including human cardiac myosin (HCM). Monoclonal antibodies were raised against HCM by hybridoma technique. Antimyosin antibody producing clones were identified by ELISA and monoclonality was established by limiting dilution. The antibodies were purified, isotyped and their cross reactions with myosin from other species were estimated. All the clones showed negligible cross reaction with rabbit myosin, but reacted with bovine skeletal myosin to different extents (40-100%). The most avid antibody Mab 4G4 which also strongly reacted with rat cardiac myosin, was labelled with 125I using different oxidising agents such as iodogen, chloramine-T and lactoperoxidase. More than 95% pure radiolabelled antibody could be obtained by gel filtration. The immunoreactivity was retained. Mab 4G4 was also labelled with 99mTc using stannous tartrate as the reducing agent. Radiolabelling yield was approximately 60%, the purity was >95%. Both the radiolabelled preparations were tested for biodistribution in rats--both normal and those with induced MI. Approximately 0.7 % of the injected activity/g was found in the infarcted region and the accumulation of activity in the infarcted heart was 1.5 times that in the normal heart. A very high percentage of activity (80%) accumulated in the thyroid. With further optimisation of labelling and use of F(ab')2 fragments, better delineation of the infarct sites may become possible. PMID- 12562014 TI - Evaluation of rabies virus neutralizing antibody titres induced by intramuscular inoculation of rabies DNA vaccine in mice and Bonnet monkeys (Macaca radiata). AB - A rabies DNA vaccine consisting of plasmid DNA expressing the rabies virus surface glycoprotein was injected (im) twice at two week interval to outbred swiss mice or Bonnet monkeys (Macaca radiata) and the levels of rabies virus neutralizing antibody (VNA) titres were examined over a one year period. In mice, the VNA titre was maintained above the minimum protective level (0.5 I.U./ml) up to 10 months after primary immunization, while in monkeys, the titre dropped below the protective level by 6 months. An anamnestic B cell response was seen in both mice and monkeys following the administration of a booster dose, 10 and 6 months after the primary immunization, respectively. These results indicate that im injection of rabies DNA vaccine induces VNA in nonhuman primates and mice unlike intradermal (id) immunization, which was shown to induce VNA only in mice but not in monkeys. This is the first report on the induction of VNA in nonhuman primates by im inoculation of rabies DNA vaccine. PMID- 12562015 TI - Studies on digoxin--14C-acetate incorporation in to digoxin and degenerative changes in the brain in rats administered digoxin. AB - The human hypothalamus produces an endogenous membrane Na+-K+ ATPase inhibitor digoxin. Digoxin is a steroidal glycoside and could be synthesised by the isoprenoid pathway. The other metabolites of the isoprenoid pathway are cholesterol, dolichol and ubiquinone. We have tried to find out the extent of incorporation of 14C acetate into digoxin in rat brain. The effects of digoxin administration on the rat brain was also studied. The results show that the percentage incorporation of 14C acetate into digoxin is low but detectable. The maximum incorporation was observed for cholesterol, followed by dolichol and finally ubiquinone. The histopathological changes observed after digoxin administration were focal degeneration of the ganglion cells in the cerebrum and cerebellum. The carbohydrate components of the glycoproteins were reduced and the concentration of serotonin, dopamine, and epinephrine showed a significant increase. The role of digoxin in mediating neuronal cell death is discussed. PMID- 12562016 TI - Voltage activated calcium channels in somatic muscle of filarial nematode Setaria cervi. AB - Acetylcholine (Ach), levamisole and pyrantel pamoate all cause stimulation of spontaneous rhythmic movements of whole worm and nerve muscle preparation of filarial nematode Setaria cervi. These stimulant effects are manifested only in the presence of available Ca2+ or extracellular Ca2+. Electrical stimulation of nerve muscle preparation of Setaria cervi elicited depolarization and increase in amplitude and tone of contractions. Electrical current stimulates Ca2+ entry leading to depolarization and during the phase of depolarization addition of any of the three stimulants viz. Ach, levamisole or pyrantel pamoate fails to elicit any response on nerve muscle preparation. The findings indicate that electrical stimulation, excitatory neurotransmitter Ach and stimulant anthelmintics levamisole and pyrantel pamoate all produce their stimulant effect by triggering entry of Ca2+ into the muscle cell. Further, blocking the calcium channels by nifedepine and thereby the entry of Ca2+ into the cells blocks the stimulant effect of Ach levamisole and pyrantel pamoate. PMID- 12562017 TI - Protection against rotavirus diarrhoea in mice by trypsin inhibitor. AB - To investigate the role of soyabean trypsin inhibitor (TI) during rotavirus (RV) diarrhoea, changes in enzyme activities of six relevant mucosal enzymes (lactase, sucrase, maltase, trehalase, glucoamylase and alkaline phosphatase) were assayed following inoculation of suckling mice with EB rotavirus (serotype 3) along with the TI and compared with the age-matched healthy control mice. The animals were divided into three groups i.e. group 1 (controls), group 2 (RV inoculated) and group 3 (RV + TI inoculated and sacrificed under light anaesthesia on 0, 1, 3, 5, 7 and 10 day post inoculation (dpi). Then intestines were excised and divided into two parts (jejunum and ileum). They were separately homogenized in 0.9% cold normal saline and activities of mucosal enzyme were measured. Alkaline phosphatase and disaccharidases were found to be decreased significantly in RV inoculated animals in both the anatomical portions of small intestine of mice. These enzyme levels were restored with the administration of TI i.e. in group 3 and became comparable to the controls in both intestinal portions. These studies suggest that activity of intestinal enzymes which are important in digestive absorptive functions of small intestine were restored with the addition of TI whengiven to infant mice showing its protective efficacy during rotavirus infection. PMID- 12562018 TI - Differential effect of polyherbal, antiobesity preparation, OB-200G in male and female mice and monosodium glutamate-treated rats. AB - Effect of administration of different doses (0.25, 0.5, 1 and 2 g/kg, twice daily, po) of a polyherbal preparation, OB-200G and fluoxetine (10 mg/kg, ip) for 21 days was studied on food intake and body weight in male and female Laka mice. The study further investigated the effect of administration of 0.5 g/kg dose of OB-200G for 40 days on body weight, fat pad weights, locomotor activity and biochemical parameters in monosodium glutamate (MSG)-treated male and female Wistar rat pups. Administration of OB-200G produced dose dependent decrease in body weight in both male and female mice. On the other hand, fluoxetine decreased body weight only in female mice. The food intake was significantly (P < 0.05) increased in both fasted male and female mice after treatment with the lower dose (0.25 g/kg, po) of OB-200G. However, significant (P < 0.05) decrease in food intake was recorded with the administration of higher doses (0.5, 1 and 2 g/kg, po) of OB-200G and fluoxetine in fasted female mice on day 1, 7, 14 and 21. But in male mice differential effect on food intake was recorded at different doses on day 1, 7, 14 and 21. Further, OB-200G administration significantly (P < 0.05) decreased body weight and fat pad weights, increased serum glucose levels and ambulatory activity in MSG-treated female rats but not in MSG-treated male rats. The results suggest that OB-200G involves gender differences in mediating its antiobesity effect and may supplement the current armamentarium for the treatment of obesity. PMID- 12562019 TI - Effects of photodynamic treatment on biological antioxidant systems in rats. AB - Effects of photodynamic treatments on inherent antioxidant metabolites and cellular defence enzymes have been investigated in rats. Wistar rats were grouped into untreated controls, light controls, hematoporphyrin derivative (Hpd) (treated with 5 and 10 mg Hpd/kg body weight and kept in dark) and sets treated with both Hpd and red light (dose 172 and 344 j/m2 ). After 2, 24, 48 and 72 hr of Hpd injection the rats sacrificed, livers quickly excised to analyze Hpd uptake, activities of enzymes like catalase, GSH-Px and antioxidants like GSH, vitamin A, vitamin E and vitamin C. The results showed that the loss of Hpd from liver as a function of post- injection time was non- linear. An increased generation of lipid radicals was observed in the groups treated with 5 mg Hpd and higher dose of light and in groups treated with 10 mg Hpd at both the doses of light. Combination of light and Hpd reduced hepatic GSH content with a concomitant reduction in GSH-Px. At higher doses of Hpd and light, there was a significant reduction in hepatic vitamin A levels. Combination of Hpd and light in all doses reduced vitamin E content in liver. The decreased biological antioxidant contents and GSH-Px may be attributed to their utilization for the scavenging of free radicals generated by Hpd and light in tissues. However, no change in catalase activity and vitamin C content in liver was noted in experimental rats. The results suggest that exposure to higher doses of Hpd with light alters oxidant stress system and TBARS content in rat. PMID- 12562020 TI - Permeabilization and in situ adsorption studies during growth and coumarin production in hairy root cultures of Cichorium intybus L. AB - Effect of addition of a permeabilizing agent dimethyl sulfoxide (DMSO) and a solid adsorbent, XAD -7, on growth and coumarin production in hairy root cultures of C. intybus was studied. Continuous permeabilization of the hairy root cultures of C. intybus with DMSO has been shown to be an effective strategy for enhanced release of coumarins while preserving the root viability. DMSO at 0.2% (v/v) level showed the maximum growth and coumarin production but was less as compared to control on day 28. Treatment of cells with increasing concentrations of DMSO (0.3 - 0.6 % v/v) to hairy root cultures of C. intybus, showed an inverse relationship with growth and coumarin production. Growth and production of coumarins increased with 1% media filtrate (MF) of cultures of Phytopthora parasitica var. nicotiana treatment. It was observed that treatment with DMSO (0.2% v/v) and 1% MF of P. parasitica showed the better growth and coumarin production with an increased release of coumarins as compared to the control and other treatments. It was observed that treatment of hairy root cultures with XAD 7 resulted in lesser growth and coumarin production as compared to control during the culture period. Addition of XAD-7 along with 1% MF of P. parasitica showed enhanced growth, coumarin production and increased adsorption as compared to control and lone XAD-7 treatment. Combined addition of DMSO/XAD-7 with fungal elicitor showed synergistic response in terms of biomass and coumarin production. Excretion of coumarins in both the cases was dependent on the presence of DMSO/XAD-7. These results showed that continuous permeabilization of hairy root cultures of C. intybus by using DMSO at 0.2% (v/v) level coupled with 1% MF of P. parasitica maintained viability of tissues and produced coumarins at higher level. PMID- 12562021 TI - Growth, CO2 exchange rate and dry matter partitioning in mungbean (Vigna radiata L.) grown under elevated CO2. AB - This study was conducted to determine the effects of anticipated future level of CO2 on growth and dry matter partitioning of mungbean (Vigna radiata). Plants were grown from seedlings to maturity inside the open top chamber under amhient CO2 (350 +/- 25 microL L(-1)) and elevated CO2 (600 +/- 50 microL L(-1)) at Indian Agricultural Research Institute, New Delhi (India). Plants were harvested at 20, 35 and 50 days after germination. Mungbean plants grown under elevated CO2 concentration resulted in greater photosynthetic rate on a leaflet area basis and no acclimation in photosynthesis was recorded due to high CO2. Plants grown under CO, enrichmcnt were taller and attained greater leaf area along with more dry matter than ambient CO2 grown plants at all growth stages. Response to high CO, depends upon the growth stage of the plant and it was more at early growth stages compared to maturity stages. The high CO2 grown mungbean plants also exhibited increased root growth along with stem and leaves. There was a substantial increase in pod number and seed number/plant under elevated CO2 conditions. The increase in dry matter and growth of root, stem and leaves proved that CO2 enrichment of the atmosphere can stimulate photosynthetic rate which can ultimately lead to an increase in dry matter and growth. PMID- 12562022 TI - A study of extracellular alkaline protease from Bacillus subtilis NCIM 2713. AB - An alkaline protease was isolated from culture filtrate of B. subtilis NCIM 2713 by ammonium sulphate precipitation and was purified by gel filtration. With casein as a substrate, the proteolytic activity of the purified protease was found to be optimal at pH 8.0 and temperature 70 degrees C. The purified protease had molecular weight 20 kDa, Isoelectric point 5.2 and km 2.5 mg ml(-1). The enzyme was stable over the pH range 6.5-9.0 at 37 degrees C for 3 hr. During chromatographic separation this protease was found to be susceptible to autolytic degradation in the absence of Ca2+. Ca2+ was not only required for the enzyme activity but also for the stability of the enzyme above 50 degrees C. About 62% activity was retained after 60 min at pH 8.0 and 55 degrees C. DFP and PMSF completely inhibited the activity of this enzyme, while in the presence of EDTA only 33% activity remained. However, it was not affected either by sulfhydryl reagent, or by divalent metal cations, except SDS and Hg2+. The results indicated that this is a serine protease. PMID- 12562023 TI - 14C-[lignin]-lignocellulose biodegradation by bacteria isolated from polluted soil. AB - Four bacterial species [Branhamella catarrhalis (gram -ve), Brochothrix species (gram -ve), Micrococcus luteus (gram +ve) and Bacillus firmus (gram +ve)], isolated from the soil polluted with cane sugar factory effluents, were found capable of growing on solid media supplemented with indulin AT (a polymeric industrial lignin) as sole C source. All the four species could metabolize cinnamic acid (a non-hydroxylated phenylpropanoid) as sole carbon source with significant suppression on addition of readily metabolizable carbon source (glucose). However, Br. catarrhalis and Brochothrix sp. were capable of metabolizing ferulic acid, but could not do so on addition of glucose. Of the four species, Br. catarrhalis could evolve significant amount of 14CO2 from U-14C (lignin)-lignocellulose prepared from rice stalks (ca. 10% of the added radioactivity in 3 weeks), in addition to solubilization of another 11.7% radioactivity in culture filtrate. The other three species could not significantly evolve 14CO2, though a significant fraction of added 14C-lignin (6.1 to 11.2%) could be solubilized into culture filtrate, suggesting lack of ring-cleavage or other CO2 evolving mechanisms in these species. PMID- 12562024 TI - Removal of copper by Pseudomonas putida strain S4 isolated from copper mines. AB - A bacterial strain, Pseudomonas putida S4, was isolated from smelter drainage of copper mines. The strain exhibited resistance to several heavy metals, like aluminium (Al), zinc (Zn), nickel (Ni), cobalt (Co) besides copper (Cu). Strain S4 could accumulate Cu from the Cu-supplemented growth medium. In the present study, we have demonstrated the Cu2+ removal capacity of this strain from various samples such as mine effluent, low-grade ore and ore-tailings, collected from the mining site. Moreover, approximately 80% of the accumulated Cu2+ could be recovered from the loaded biomass by a simple desorption procedure. PMID- 12562025 TI - Micropropagation of Paulownia fortuneii through in vitro axillary shoot proliferation. AB - Primary cultures were established with nodal segments from juvenile shoots of two year-old Paulownia fortuneii trees from a clonal plantation in Andhra Pradesh. A medium containing half-strength MS salts + RAP (1 mg/L) + sucrose (2%) produced optimum bud break in nodal explants. The same basal medium with reduced hormone level (0.5 mg/L) supported maximum multiplication of secondary cultures of P. fortuneii (1:6 in 6 weeks). Specific treatments were tested to enhance this rate of multiplication. In one approach, five to six week old in vitro grown shoots were ratooned (cutting the main shoot at the bottom leaving one node). The stumps (ratooned basal node) produced 2 to 3 axillary shoots, which grew into 4 to 5 nodes by 3 weeks; thus, providing additional shoots from the same explant. This provided 30% additional shoots in 4 cycles. Secondly, reducing the light intensity to 1200 lux resulted in higher shoot elongation, i.e, formation of 8 nodes in 5 weeks with healthier shoots than the normal intensity of 3000 lux under which only 6 nodes were produced in 6 weeks. In vitro-grown shoots could be successfully rooted ex vitro in vermiculite + cocopeat mixture (1:1 v/v) under 90% humidity, transferred to soil in polybags for hardening in the green house for 2 weeks and shifted to shade net for further hardening. After one month, the plants could be successfully transplanted to field with 95% survival. Micropropagated plants showed an excellent growth in the field attaining a height of 1.5 m and a collar diameter of 2.8 cm in 3 months. PMID- 12562026 TI - A systemic resistance inducing antiviral protein with N-glycosidase activity from Bougainvillea xbuttiana leaves. AB - An antiviral protein from Bougainvillea xbuttiana leaves induced systemic resistance in host plants N. glutinosa and Cyamopsis tetragonoloba against TMV and SRV, respectively which was reversed by actinomycin D, when applied immediately or shortly after antiviral protein treatment. When the inhibitor was applied to the host plant leaves post inoculation, it was effective if applied upto 4 h after virus infection. It also delayed the expression of symptoms in systemic hosts of TMV. The inhibitor showed characteristic N-glycosidase activity on 25S rRNA of tobacco ribosomes, suggesting that it could also be interfering with virus multiplication through ribosome-inactivation process. PMID- 12562027 TI - JU-2, a novel phosphorous-containing antifungal antibiotic from Streptomyces kanamyceticus M8. AB - A novel phosphorous-containing antifungal antibiotic JU-2 was isolated from Streptomyces kanamyceticus M8. Quantitative chemical analysis shows the presence of two phenylalanines, two glucose, one linoleic acid, one crucic acid and one phosphonamide moiety per molcule of the antibiotic. JU-2 shows strong inhibitory activity against various pathogenic and non-pathogenic fungi but no activity against bacteria and yeast. PMID- 12562028 TI - Adherence to antiretroviral therapy: where are we, and where do we go from here? PMID- 12562029 TI - Adherence to antiretroviral therapy among patients with HIV: a critical link between behavioral and biomedical sciences. AB - Rapid advances in biomedical science, such as pharmaceutical developments for HIV disease, must be integrated with advances in behavioral science to further our understanding of medication adherence. This article evaluates the current state of the science in adherence to antiretroviral therapy for persons with HIV. The primary objectives are to 1) identify critical determinants of adherence, and 2) describe interventions to improve adherence. Adherence is a complex dynamic behavior influenced by characteristics of the patient, treatment regimen, disease, patient-provider relationship, and clinical setting. Therefore, the most promising interventions are multifaceted and target different locations in this matrix simultaneously. Unfortunately, nonadherence remains a formidable barrier in the management of HIV, resulting in the development of resistance and drug failure. Moreover, adherence is a public health concern, with implications for the transmission of HIV in general and the transmission of drug-resistant strains of HIV specifically. Despite substantial attention to adherence in recent years, much more remains to be done to better understand and promote adherence to antiretroviral therapy through effective interventions. From this integration of biomedical and behavioral science, effective clinical interventions can be developed and implemented to enhance the health of patients with HIV. PMID- 12562030 TI - Measurement of adherence to antiretroviral medications. AB - Measurement of adherence may be important in determining why patients fail antiretroviral therapy. Although patient self-report is by far the most frequently used means of assessing adherence, it overestimates adherence. However, patients who state they are nonadherent almost always are. The pill identification test is a recently described tool that may be useful in clinical practice. The best methods of adherence measurement are pill counts and electronic monitoring. Pill counts suffer from inability to record the time of consumption of therapy. Electronic monitoring enables timing of pill consumption and is the closest to a gold standard for measuring adherence. However, this is only the case if patients are carefully instructed in how to use the device, e.g., not to remove extra doses from their pill bottle. A composite adherence score has been developed that uses electronic monitoring, pill counts, and patient self-report. The authors believe that careful measurement of adherence is essential in the assessment of a patient failing to respond to antiretroviral therapy. PMID- 12562031 TI - Computer-assisted self-interviewing (CASI) to improve provider assessment of adherence in routine clinical practice. AB - Adherence to HIV antiretroviral therapy is closely associated with viral load suppression, progression to AIDS, and death in HIV-infected people. There is, however, no standardized approach to adherence assessment in routine clinical practice, and several studies suggest that providers rarely estimate adherence better than chance. Computer-assisted self-interview technology is an efficient means to identify HIV antiretroviral regimen errors and monitor adherence in order to maximize the benefits of therapy. PMID- 12562032 TI - Virologic and immunologic response, clinical progression, and highly active antiretroviral therapy adherence. AB - A growing body of evidence suggests that a high degree of adherence is required to achieve and maintain a successful virologic response both in the short and long term. This holds true despite the definition of adherence or how it is measured. Reported differences in the degree of adherence required are likely due to differences in study design, difficulty measuring patient adherence, patient population studied, and the antiretroviral regimen studied. Virologic suppression and immunologic response often go hand in hand, but the impact of adherence on change in CD4 count tends to be delayed and, therefore, less apparent than the impact on HIV viral load. Degree of adherence has also been shown to be associated with AIDS-related morbidity, mortality, and hospitalizations. PMID- 12562033 TI - Virologic correlates of adherence to antiretroviral medications and therapeutic failure. AB - Adherence to antiretroviral therapy affects the pharmacokinetics of antiviral drugs and activates a cascade of events ultimately leading to therapeutic success or failure. An optimal adherence usually affords minimal rounds of virus replication and rare spontaneous mutations, which are unable to be fixed in the genome because of the competition of wild-type (more fit) strains. Therefore, adherence-based therapeutic success is mostly accompanied by the prevalence of wild-type strains. In case of poor adherence, virus replication is substantial, and mutations randomly occurring tend to be fixed within the genome. Under these conditions, mutated-resistant strains will outgrow wild-type virus (sensitive to antivirals and thereby unable to compete enough with resistant strains for cellular targets): thus, therapeutic failure occurs, and mutated resistant strains are predominant. In the case of very low or absent adherence, virologic failure occurs, although wild-type virus (whose replication is not significantly affected by antivirals) is not outgrown by mutated strains randomly produced but unable to be fixed within the genome. Taken together, these events and their consequences strongly support the relevance of a tight and continuous monitoring of adherence to antiretroviral drugs to prevent the risk of development of mutated strains often cross-resistant to the majority of antiretroviral drugs currently available. PMID- 12562034 TI - Correlates and predictors of adherence to highly active antiretroviral therapy: overview of published literature. AB - Knowledge of factors associated with medication adherence could help HIV clinicians to target persons in need of intervention, design these interventions, and help researchers to plan studies of adherence. This review summarizes the results of 20 studies investigating the issue of barriers to optimal highly active antiretroviral therapy (HAART) adherence. Only a few determinants were consistently associated with nonadherence. Symptoms and adverse drug effects, psychologic distress, lack of social or family support, complexity of the HAART regimen, low patient self-efficacy, and inconvenience of treatment were the factors most consistently associated with nonadherence. There were inconsistent findings regarding the relationship of adherence and the following variables: sociodemographic characteristics, substance abuse, depressive symptoms, quality of life, CD4+ cell count, knowledge and beliefs about treatment, patients' satisfaction with health care, and patient-provider relationship. A synthesis of findings relating various factors to adherence to HAART is difficult to reach because of several limitations of the existing body of research. These limitations concern the measurement of adherence, the assessment of correlates and predictors of adherence, the study population, and the study design. PMID- 12562035 TI - Treatment-related factors and highly active antiretroviral therapy adherence. AB - Adherence to highly active antiretroviral therapy (HAART) plays a critical role in the effectiveness of HIV treatment. Nevertheless, the complexity of regimens and frequent side effects make HAART extraordinarily difficult to take, and many HIV-infected persons fail to adhere. The current study offers an overview of the relationship between adherence and antiretroviral treatment-related variables. As for other chronic diseases, medication regimen complexity also has an impact on adherence in the management of HIV infection. In particular, the authors discuss the effect of pill burden, dosing frequency, dietary instructions, number and type of different medications prescribed, short- and long-term side effects, convenience, and ability to incorporate the treatment regimen into a daily routine. Medication side effects are common in HAART-treated persons and are associated with concurrent and future nonadherence. Simplification of regimens, adjustment of the drug schedule to the patient's specific lifestyle, and anticipation and self-management of side effects are treatment-based strategies to optimize HAART adherence and ensure the most effective, convenient, safe, and well-tolerated antiretroviral treatment. PMID- 12562036 TI - Neurocognitive aspects of medication adherence in HIV infection. AB - Strict adherence to the current highly active antiretroviral therapies (HAART) is a prerequisite for treatment success. The complexities of the treatment regimens places high demands on motivational and cognitive factors, and studies of cognitive predictors of adherence are just beginning to emerge. Traditional measures of new learning and recall may not be among the best neuropsychologic predictors of adherence. Rather, certain aspects of frontal and executive functioning appear to be more strongly associated with good adherence. PMID- 12562037 TI - Depression is a risk factor for suboptimal adherence to highly active antiretroviral therapy. AB - Affective disorders have been reported as the most common mental health problem in persons with HIV infection. Depression has a significant impact on the quality of life of persons living with HIV and AIDS and is associated with HIV disease progression and mortality, even after controlling for sociodemographic and clinical characteristics and substance abuse. Depression has been also reported as one of the main causes of poor adherence with antiretroviral regimens. However, no published investigation has specifically focused on the relationship between depression and adherence to antiretroviral therapy. Nonetheless, information on the association between depressive symptoms and adherence may be gathered from investigations carried out to explore determinants of adherence with antiretroviral therapy. Findings from available studies show a substantial and consistent relationship between adherence to antiretroviral regimens and depression. Early recognition and proper management of depressive comorbidity could be an effective intervention strategy to improve adherence and may make a difference in the quality of life, social functioning, and disease course of people with HIV. PMID- 12562038 TI - Relationship between HAART adherence and adipose tissue alterations. AB - Adipose tissue alterations (ATA), which are common among persons treated with highly active antiretroviral therapy (HAART), can have substantial psychologic repercussions, with a subsequent negative impact on the patient's quality of life and on HAART adherence. However, the cross-sectional nature of the studies precludes establishing the direction and causality of the relationship. The authors evaluated the longitudinal relationship between ATA and adherence to HAART. The analysis included all participants in the AdICoNA and the LipolCoNA substudies of the Italian Cohort Naive Antiretrovirals (ICoNA). Adherence was assessed using a 16-item self-administered questionnaire, which also included a question on self-perceived fat accumulation experienced during the past 4 weeks. ATA was diagnosed by physicians at enrollment and evaluated every 6 months thereafter. There were 207 patients, with a median age of 35 years; 73% were men; and 34% acquired HIV through injection drug use. At baseline, nonadherence was reported by 63% of participants, and ATA was self-perceived by 15% and clinically diagnosed in 25%. Using Cox regression analysis, patients with good adherence at baseline were more likely to develop ATA (RH = 2.58; 95% CI, 1.09-6.11) and developed it sooner. Self-perceived ATA at baseline was independently related to subsequent nonadherence (OR, 4.67; 95% CI, 1.01-22.4), but clinically diagnosed ATA was not (OR, 0.77; 95% CI, 0.37-1.61). Patients' adherence to HAART is a dynamic process that interacts with ATA. Better adherence is associated with a higher risk of subsequent occurrence of ATA, while patient-perceived onset of morphologic alterations can reduce adherence to antiretroviral therapy. PMID- 12562039 TI - Behavioral correlates of adherence to antiretroviral therapy. AB - The objective of this study was to analyze the relationships between adherence to treatment and sexual and drug-taking behaviors among persons with HIV, who started combination antiretroviral therapy as their first regimen. The authors analyzed data from 366 patients enrolled in a multicenter observational cohort study conducted in infectious disease hospital units in Italy. Adherence measurement was based on responses to a self-administered questionnaire regarding following HIV physician advice on taking medications and missed appointments. Questions on sexual and drug-taking behaviors were also included in the questionnaire. The median time since starting antiretroviral therapy was 11.8 months; 37.4% of patients were on a two-drug regimen and 62.6% were on a three drug regimen. Overall, 68 patients (18.6%) could be classified as nonadherent. The proportion of patients with viral load < or = 500 copies/mL was significantly higher among adherent patients (68%) compared with nonadherent patients (40.4%; p = .001). In multivariable analysis, age (odds ratio [OR], 0.65; 95% confidence interval [CI], 0.42-0.98, per 10-year increment) and current use of injection (OR, 3.47; 95% CI, 1.40-8.5) or noninjection drugs (OR, 4.23; 95% Cl, 1.85-9.67) were significantly associated with nonadherence. No significant association was found between adherence and sexual behaviors. The data do not support the hypothesis that among HIV-infected person on antiretroviral therapy, poor adherence is associated with high-risk sexual behaviors that may further spread the infection. PMID- 12562040 TI - Nonadherence among HIV-infected injecting drug users: the impact of social instability. AB - The authors tested the impact of social instability on adherence to highly active antiretroviral therapy (HAART) among patients infected with HIV through injection drug use (IDU; MANIF2000 cohort). In the study, they analyzed sociodemographic baseline characteristics to develop an indicator of social instability. Information concerning adherence to HAART was collected through questionnaires during a 2-year follow-up period. Factors associated with nonadherence were studied in two different groups: 1) patients who had stopped injection drug use (ex-IDUs) and who were not in drug maintenance programs (DMT) during the entire follow-up period, and 2) those who were still opiate dependent. Among the 210 eligible patients, 114 were classified as ex-IDUs and 96 as opiate dependent. Ex IDUs reported nonadherence behaviors in 96 of 384 visits (25.0%), while opiate dependent patients were nonadherent in 111 of 308 visits (36.0%; p = .02). Among ex-IDUs, the only factor associated with nonadherence was social instability, while among opiate-dependent patients, injection behavior was the only determinant of nonadherence behavior. For opiate-dependent patients, DMT may enhance adherence to HAART, but only if it is successful in reducing abuse of injection practices. For ex-IDUs, it is very important that the management of social difficulties be taken into account to increase adherence to HAART. PMID- 12562041 TI - Interventions to improve adherence to antiretroviral therapy. AB - Recent research has shown that very high levels of adherence are required to obtain the maximum benefit of highly active antiretroviral therapy (HAART). This situation justifies the importance of developing efficient strategies to improve adherence to drugs against HIV. A comprehensive range of factors are the targets of the most effective interventions to improve adherence, including cognitive, behavioral, emotional, and social aspects. The authors describe three theoretical models that can help identify barriers and guide interventions. Most of the interventions are complex and may include more convenient care, provision of information, counseling, reminders, reinforcement, self-monitoring, family therapy, or additional supervision or attention. The authors suggest aspects that should be included in interventions to promote adherence based on a review of the literature and clinical experience. Limited evidence suggests that interventions to enhance adherence to antiretroviral therapy in people with HIV are most likely to be successful when they are comprehensive, longitudinal, and tailored to the person. PMID- 12562042 TI - Physician estimates of adherence and the patient-physician relationship as a setting to improve adherence to antiretroviral therapy. AB - Physicians estimate their patients' adherence to medications, and base decisions about treatment on these estimates. In HIV, misjudgment of patient adherence can have adverse consequences, including withholding of therapy, unnecessary changes in therapy, or unnecessary laboratory testing. A review of the literature demonstrates that physicians are often inaccurate in estimating patient adherence with antiretroviral therapy. These findings have implications for practice. Standardized methods for adherence assessment are currently available that can be used to enhance physicians' ability to understand adherence behavior and barriers. The patient-physician relationship presents a unique setting for improving adherence. The authors propose interventions to improve adherence within the context of the patient-physician relationship at the physician level, interpersonal level, and organizational level. Improved communication, including discussion about patient lifestyle and preferences, can facilitate a frank exchange of information, negotiation, and a spirit of cooperation. Active patient participation in the decision-making process is crucial. PMID- 12562043 TI - Chronic disease self-management and adherence to HIV medications. AB - Although it is widely recognized that potent antiretroviral therapies have transformed HIV from an acute to a chronic illness, it is less often recognized that with this change, a different model of care is appropriate. People living with chronic HIV must function independently, taking personal long-term, day-to day responsibility for care. The role of doctors and the healthcare system is to help them do this. The authors present a conceptual framework for the self management of chronic HIV disease and discuss the components of an HIV self management program. The ability to take antiretroviral medications well and consistently is a key part of self-management but should be nested within an array of important skills, including symptom-management skills, goal setting and planning, communication, and accessing information and resources. PMID- 12562044 TI - Highly active antiretroviral therapy adherence: the patient's point of view. AB - There is a widespread acknowledgment that successful treatment with highly active antiretroviral therapy (HAART) requires the patient to maintain consistent adherence to the prescribed regimen on a long-term basis. Adherent patients have been shown to have reduced viral loads and increased CD4 counts, live longer, and have better quality-adjusted survival. The consequences of nonadherence are also obvious: poorer adherence leads to inadequate viral suppression and continued disease progression. With the astonishing advances in medical therapeutics during the past two decades, one may expect a similar proliferation of studies of the nature of nonadherence and tests of strategies to help patients overcome it. However, the literature and the know-how concerning interventions to improve adherence are still surprisingly weak. The unique challenges posed by combination therapies for HIV and AIDS require healthcare providers to address the issues created by adherence to complex regimens through multifactorial interventions tailored to patients' needs. PMID- 12562045 TI - Mechanisms of bisphosphonate effects on osteoclasts, tumor cell growth, and metastasis. AB - Bisphosphonates are potent inhibitors of osteoclast-mediated bone resorption that also exhibit antitumor activity. There is now extensive in vitro evidence that bisphosphonates inhibit proliferation and induce apoptosis of tumor cell lines. In addition, they appear to inhibit tumor cell adhesion and invasion of the extracellular matrix. These data are supported by a growing body of evidence from animal models demonstrating that bisphosphonates can reduce skeletal tumor burden. This may reflect direct antitumor effects or indirect effects via osteoclast inhibition and alteration of the bone microenvironment. Research has begun to shed light on the complex mechanisms by which bisphosphonates inhibit bone resorption and interfere with the formation and growth of bone lesions. Nitrogen-containing bisphosphonates inhibit protein prenylation and thereby short circuit intracellular signaling via small guanine triphosphatases, such as Ras, which require membrane localization. As a result of these biochemical effects on the mevalonate pathway, bisphosphonates appear to modulate the expression of bcl 2 leading to caspase-dependent apoptosis, inhibit matrix metalloproteinases, downregulate alphavbeta3 and alphavbeta5 integrins, and increase expression of osteoprotegerin, thereby antagonizing osteoclastogenesis. Further preclinical studies are ongoing to fully elucidate these biochemical mechanisms, and well designed clinical trials are necessary to investigate whether the antitumor potential of bisphosphonates can be realized in the clinical setting. PMID- 12562046 TI - Efficacy of bisphosphonates in the management of skeletal complications of bone metastases and selection of clinical endpoints. AB - Bisphosphonates are the current standard of palliative care for patients with bone lesions from breast cancer and multiple myeloma. This article discusses the selection of endpoints and statistical methods used to assess clinical efficacy of bisphosphonate therapies in patients with bone metastases. Recent studies of pamidronate and zoledronic acid have set the standards for the design and conduct of multicenter, randomized, placebo-controlled trials to assess the clinical benefit of bisphosphonates in patients with bone metastases. Studies of zoledronic acid have demonstrated objective, significant, and enduring benefits in patients with a broad range of primary cancers, including prostate cancer, using robust clinical endpoints. Other bisphosphonates, including clodronate, have been investigated for the treatment of bone metastases, but these studies have been relatively small. This review first considers issues of trial design and analysis, with particular emphasis on the statistical requirements for the rigorous analysis of multiple events occurring in the same patient, and then reviews the results of bisphosphonate trials in patients with breast or prostate cancers metastatic to bone in the light of these statistical considerations. PMID- 12562047 TI - Broad clinical activity of zoledronic acid in osteolytic to osteoblastic bone lesions in patients with a broad range of solid tumors. AB - Bone metastases are a common feature of a variety of solid tumors and are associated with substantial skeletal morbidity, including severe bone pain and pathologic fractures. Treatment with bisphosphonates, primarily pamidronate, is the current standard of care for patients with breast cancer and multiple myeloma who have predominantly osteolytic lesions. However, until recently no bisphosphonate had demonstrated efficacy in patients with osteoblastic lesions, which are common during the progression of prostate cancer and other solid tumors. Zoledronic acid, a potent, new-generation, nitrogen-containing bisphosphonate, has demonstrated significant benefits for patients with bone metastases resulting from a broad range of primary tumors, including multiple myeloma and breast, lung, kidney, and prostate cancers, and other solid tumors. Benefits include a decreased incidence of pathologic fractures and longer time to the first skeletal complication. Zoledronic acid is the first and only bisphosphonate to be proved effective in patients with all types of bone lesions, from osteolytic to osteoblastic, and therefore represents an important therapeutic advancement in the treatment of bone metastases. PMID- 12562049 TI - Future directions in the treatment and prevention of bone metastases. AB - Preclinical studies are providing a growing body of evidence that bisphosphonates, particularly nitrogen-containing bisphosphonates, have antitumor activity. Bisphosphonates induce tumor cell apoptosis and reduce skeletal tumor burden in tumor xenograft models. Clinical studies with daily oral clodronate suggest that bisphosphonates can prevent bone metastases when used in the adjuvant setting, but the effect on overall survival is less certain. The more potent nitrogen-containing bisphosphonates, i.e., pamidronate and zoledronic acid, have demonstrated antitumor activity at approximately 10- to 100-fold lower concentrations than clodronate in vitro. A number of important unanswered questions must be addressed regarding the optimal use of bisphosphonates for prevention of bone metastases. For example, when should treatment begin, how long must treatment be continued, and what are the optimal dose and schedule to achieve clinically meaningful antitumor effects? Adjuvant studies of zoledronic acid in patients with breast and prostate cancer are under development, and the results are eagerly anticipated. PMID- 12562048 TI - Efficacy of zoledronic acid and pamidronate in breast cancer patients: a comparative analysis of randomized phase III trials. AB - Results of a large, randomized, comparative phase III trial involving 1,130 breast cancer patients demonstrated that 4 mg zoledronic acid and 90 mg pamidronate were equally effective at reducing the occurrence and delaying the onset of skeletal-related events in patients with bone metastases secondary to breast cancer. However, zoledronic acid has demonstrated a treatment advantage over pamidronate with respect to the proportion of patients receiving radiation therapy to bone. Moreover, among patients with at least one osteolytic lesion (N = 352), zoledronic acid achieved a 17% reduction in the proportion of patients with a skeletal-related event compared with pamidronate and significantly prolonged the time to first event. In general, zoledronic acid and pamidronate had comparable effects on time to progression of bone metastases, overall disease progression, pain and analgesia scores, and overall survival. Both zoledronic acid and pamidronate effectively suppressed markers of bone resorption. However, zoledronic acid demonstrated a greater and more sustained suppressive effect. Zoledronic acid (4 mg via 15-minute infusion) also demonstrated a safety profile similar to that of pamidronate (90 mg via 2-hour infusion). In summary, the increased efficacy of zoledronic acid in specific patient subsets and the convenience of a 15-minute infusion suggest that 4 mg zoledronic acid has advantages over 90 mg pamidronate for treatment of bone metastases in patients with breast cancer. PMID- 12562050 TI - Urinary incontinence and menopause: an evidence-based treatment approach. AB - OBJECTIVE: To review the evidence on risk factors, evaluation, diagnosis, and treatments in postmenopausal women with incontinence. DATA SOURCES: Articles on urinary incontinence were located by searching Medline and the Cochrane Database of Systematic Reviews in March 2000. RESULTS: Urinary incontinence is common with aging. The relationship between urinary incontinence and menopause is not well understood. There are 3 major types of urinary incontinence: stress, urge, and mixed. The diagnosis can often be established with a good history. For a long time estrogen was thought to be of benefit in the treatment of incontinence, however, there is now evidence it may exacerbate urinary incontinence. There are multiple new agents shown to be effective in the treatment of incontinence. Surgical approaches for the treatment of stress incontinence have not been studied in large clinical trials, however, open abdominal retropubic suspension seems to be better than anterior vaginal repair. CONCLUSIONS: As the aging population grows, urinary incontinence will become an increasing public health problem. Increasing awareness is needed by women and their providers to identify, evaluate, and apply evidence-based treatment approaches to urinary incontinence. PMID- 12562051 TI - Radiographic measurements, bone mineral density, and the Singh Index in the proximal femur of white and black postmenopausal women. AB - BACKGROUND: Radiographic measurements, bone mineral density (BMD), and the Singh Index were examined to assess ethnic differences in the architecture and trabecular patterns in the proximal femur. METHODS: We measured height (cm), weight (kg), and the following radiographic variables in 326 white and black postmenopausal women participating in the Women's Health Initiative at the Clinical Center in Detroit, MI: neck breadth, inferior neck cortical thickness, head diameter, subtrochanteric breadth, and subtrochanteric medial and lateral cortical thicknesses. Bone densitometry was performed by dual-energy x-ray absorptiometry (Hologic QDR 1000 plus; Hologic Inc, Bedford, MA) at 5 regional sites in the proximal femur. The Singh Index was read by its originator. RESULTS AND CONCLUSIONS: There was no significant ethnic difference in mean age, height, or body mass index, but weight and BMD was higher in the black group at all regional sites. The inferior neck cortical thickness was significantly greater in the black group. The Singh Index was found to be grade VI (normal) in 87%, grade V in 9%, and grades II-IV in 4% of all subjects. Multiple regression models explained 35% to 60% of the variance in the regional BMDs; the Singh Index, weight, and subtrochanteric cortical thicknesses were significant contributors to all regional hip BMD models. Although there were ethnic differences in BMD, there were no ethnic differences in the distribution of the Singh Index scores. PMID- 12562052 TI - Evaluating women with chest pain for the diagnosis of coronary artery disease. AB - The evaluation of women with chest pain for the presence of important coronary artery disease may be challenging. Coronary artery disease is the leading cause of death, but it is much more prevalent in women of older age. However, chest discomfort is a very common symptom among women of all ages. Younger women are much less likely (but not completely immune) from having significant coronary disease, but the existing forms of cardiac testing are more likely to yield false results when applied to low-risk groups. Women should be assessed for their overall level of risk for coronary disease and the severity and nature of symptoms. Noninvasive cardiac testing is best applied to women at intermediate risk. There are strengths and weaknesses for each of the available imaging modalities, but both nuclear scans and echocardiography have reasonable accuracy and reliability for detecting serious coronary disease in women. Importantly, a negative study imparts an excellent prognosis. The application of a systematic and comprehensive evaluation of women will improve detection of important coronary disease and permit reassurance to women who do not have evidence for ischemia producing coronary disease. For women whose symptoms are determined not to be caused by significant coronary disease, additional evaluation for noncardiac causes can then be undertaken. PMID- 12562053 TI - Subclinical hypothyroidism in women: who to treat. PMID- 12562054 TI - Herbs commonly used by women: an evidence-based review. AB - OBJECTIVE: To review the evidence of herbs commonly used by women. DATA SOURCES: Articles were located by searching Medline, Cochrane Database of Systemic Reviews, and the Combined Health Information Database and by hand searching the reference lists of recent systematic reviews. The databases were searched in January 2000 and October 2000 by using the Latin and common name of each herb. METHODS OF STUDY SELECTION: Preference was given to randomized, placebo controlled trials. When available, English language studies were reviewed. If not, data are presented from review articles that summarize the foreign study. RESULTS: Many women use herbal therapies. In the United States, herbs are considered dietary supplements. The Food and Drug Administration (FDA) cannot remove them from the market unless they are proven unsafe. The herb industry plans to improve monitoring. Many prospective randomized controlled trials are being funded. Gingko biloba seems to slow the progression of dementia but increases the risk of bleeding. St John's Wort is efficacious for treating mild to moderate depression but has many drug interactions. Ginseng seems to improve well being in perimenopausal women, but it is often impure and has side effects and drug interactions. Garlic slightly lowers blood pressure and lipids. Echinacea slightly decreases the duration of colds but does not prevent them. Valerian is beneficial for insomnia, but there is no long-term safety data. Black cohosh may help the symptoms of perimenopause, and chasteberry may improve premenstrual syndrome. More study is needed on both herbs. CONCLUSION: Some herbs are medically useful, but the American public would benefit from increased regulation. Manufacturers should be able to ensure that herbs contain pure ingredients. Side effects and drug interactions should be listed. Well-designed studies are being conducted. The results will be helpful to physicians and patients when the clinical evidence becomes available. PMID- 12562055 TI - Overview and neurobiology of attention-deficit/hyperactivity disorder. AB - Although attention-deficit/hyperactivity disorder (ADHD) impairs millions of people worldwide, both the prevalence and existence of the disorder are being reevaluated at the phenotypic level. To safeguard against overdiagnosis, the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV), demands that individuals with ADHD have pervasive impairment, that is, impairment in more than 1 setting. However, the appropriateness of the DSM-IV classification of ADHD is also undergoing reevaluation. Like the symptoms of a developmental disability, the symptoms of ADHD must be evaluated in the context of age-based norms; therefore, the current criteria for ADHD, which are not age referenced, may minimize the rate of persistence of ADHD into adulthood. In an effort to better understand the pathophysiology of ADHD, recent research has focused on identifying the etiology of ADHD. These studies have revealed that the disorder is highly heritable and may be associated with neurobiological deficits in the prefrontal cortex and related subcortical systems. Etiologic studies have also identified candidate genes and prenatal and perinatal risk factors for ADHD. As the causes and course of ADHD are better understood, a new generation of medications is being developed for the disorder. Although stimulants are often effective in reducing the symptoms of the disorder, as a class they have limitations such as a lack of 24-hour-a-day coverage, unwanted side effects, potential for abuse, and lessened effectiveness in the context of some comorbidities. Therefore, the treatment characteristics of newer, more selective treatments such as atomoxetine should continue to be explored in ADHD. PMID- 12562056 TI - Major life activity and health outcomes associated with attention deficit/hyperactivity disorder. AB - People with attention-deficit/hyperactivity disorder (ADHD) are affected by the disorder throughout their lifetimes. Children with ADHD often have comorbid oppositional defiant disorder and conduct disorder in addition to having developmental and social problems. The persistence of ADHD into adolescence and young adulthood varies according to who is being interviewed and the criteria used to define the disorder. For those adolescents and adults in whom ADHD does persist, educational difficulties continue, and problems in the areas of employment, driving, and sexual relationships emerge. ADHD is also associated with increased health care costs even when controlled for psychiatric treatment. Because most ADHD research has been conducted with male children and adolescents with ADHD, combined type, most outcomes for ADHD should be thought of as male outcomes for this subtype. In the future, ADHD researchers should study outcomes for girls and women and for people with ADHD, predominately inattentive type. PMID- 12562058 TI - Adaptive changes related to medication treatment of ADHD: listening to parents of children in clinical trials of a novel nonstimulant medication. AB - The DSM-IV diagnostic criteria for attention-deficit/hyperactivity disorder (ADHD) have proved useful in providing a common language for diagnosing, treating, and researching the disorder. Despite the utility of current ADHD diagnostic criteria, sophisticated theoretical conceptualizations of the etiology of ADHD have described a much more complex disorder that includes a range of neuropsychological impairments (such as working memory deficits and other executive dysfunction) and underlying structural and functional neuropathology (e.g., caudate nucleus volume, frontal lobe activity). Inattention, hyperactivity, and impulsivity, the hallmark triumvirate symptoms of ADHD, may be better viewed as some of the many meaningful symptoms with roots in executive functioning impairment. Outcomes of brain-imaging studies, public skepticism about diagnosis and treatment, and a demand for meaningful clinical outcomes of treatment point to a considerable need to broaden treatment-outcome criteria beyond the DSM-IV domains. The wide-ranging decrements in adaptive function and quality of life reported by parents of children diagnosed with ADHD further support core executive dysfunction. Emerging findings concerning medication related improvements in adaptive functioning (e.g., social, emotional, academic), as well as the rapid search for the neuropathology that may underlie these improvements, are fueling interest in the assessment of adaptive function in clinical trials. In a series of ongoing clinical trials of a novel nonstimulant medication for ADHD, many parents reported significant improvements in the lives of their children beyond the DSM-IV criteria. These parental reports, despite their inherent sources of error, underscore the importance of including broader and more meaningful clinical outcome assessment in clinical trials. Research protocols that omit parental interviews that assess adaptive and executive function may well overlook several meaningful and consequential medication related improvements. PMID- 12562057 TI - Novel treatments for attention-deficit/hyperactivity disorder in children. AB - Optimal medications for children with attention-deficit/hyperactivity disorder (ADHD) would be effective, well tolerated, and long acting and not cause mood swings or worsen comorbid conditions. Current medications work on brain dopamine and/or norepinephrine systems, which are thought to be involved in ADHD. The medication class with the most evidence of efficacy in ADHD is stimulants, but they may be abused, are effective for only 4 to 12 hours, and may cause mood swings or increase tic severity. In recent years, alternative treatments have been explored. Tricyclic antidepressants have efficacy comparable to that of stimulants but may cause constipation, dry mouth, tremors, blood pressure changes, and potentially serious side effects including cardiac conduction and repolarization delays. Monoamine oxidase inhibitors may improve ADHD symptoms but are associated with severe dietary restrictions. Serotonin reuptake inhibitors have little or no effect in ADHD but may improve comorbid depression. Bupropion, although less effective than stimulants, may improve both ADHD symptoms and comorbid depression. Antihypertensive agents may improve impulsivity, hyperactivity, and comorbid tics but cause sedation or rebound hypertension. Atomoxetine, which is being developed for ADHD, reduces symptoms of ADHD without exacerbating comorbid conditions and is associated with only minor side effects, including subtle changes in blood pressure and heart rate. Before prescribing a treatment, physicians should consider the appropriateness and effectiveness of any medication for children with ADHD, who may be less tolerant of side effects and less able to monitor and express concerns about their well-being than adults. PMID- 12562059 TI - Management of ADHD in adults. AB - Although first identified in children in the 19th century, attention deficit/hyperactivity disorder (ADHD) in adults was not described in the literature until 1976. The symptoms of adult ADHD resemble the symptoms of childhood ADHD, but symptom intensity, especially hyperactivity, may decrease over time. However, due to the challenges and responsibilities of adulthood, a normal day is extremely complicated for the ADHD adult. Molecular genetics and neuroimaging studies confirm that ADHD is a heterogeneous, neurobiological disorder, mainly of dopaminergic and noradrenergic pathways. Trials of pharmacologic treatments in adults with ADHD have produced mixed results due to considerable variability in diagnostic criteria, dosing, and response. This article reviews the history, neurobiology, and pharmacologic management of adult ADHD. PMID- 12562060 TI - Psychosocial treatments for attention-deficit/hyperactivity disorder in children. AB - This article provides a brief overview of the major psychosocial treatments that have some efficacy for the management of attention-deficit/hyperactivity disorder (ADHD) in children. Parent training in effective child behavior management methods, classroom behavior modification methods and academic interventions, and special educational placement appear to have the greatest promise of efficacy. Augmenting these, additional family therapy in problem-solving and communication skills and the coordination of multiple school resources across the day may be necessary. To be effective in improving prognosis, treatments must be maintained over extended periods of time. PMID- 12562061 TI - Drug development process for a product with a primary pediatric indication. AB - This article reviews the drug development process in the United States, focusing on practical issues and new U.S. Food and Drug Administration (FDA) regulations and guidance for developing a drug with a primary pediatric indication. Atomoxetine, a novel treatment for attention-deficit/ hyperactivity disorder (ADHD), is used to illustrate how the modern drug development process works and to highlight changes in the development of ADHD treatments since the introduction of the stimulants over 50 years ago. In addition to dealing with unique regulatory requirements and guidance, developing a drug for use in a pediatric population poses novel challenges in diverse areas including biomedical ethics, developmental pharmacology, and clinical trial design and implementation. PMID- 12562062 TI - Safety profile of atomoxetine in the treatment of children and adolescents with ADHD. AB - Atomoxetine is a selective norepinephrine reuptake inhibitor that is being developed for the treatment of attention-deficit/hyperactivity disorder (ADHD). Atomoxetine will be the first nonstimulant medication approved by the U.S. Food and Drug Administration (FDA) for the treatment of ADHD. Throughout the testing phases, more than 2000 children and adolescents have been exposed to atomoxetine in clinical trials, with both the number of exposures and the length of exposure time increasing. Serious adverse events have not been clearly associated with the drug, and there have been few discontinuations due to adverse events. The most common drug-related event reported in trials has been decreased appetite and an initial period of weight loss followed by an apparently normal rate of weight gain. These events tend to appear early in the course of treatment with atomoxetine and then decline. Atomoxetine has also been associated with mild increases in blood pressure and pulse that plateau during treatment and resolve upon discontinuation. There have been no effects seen on the QT interval, and the cytochrome P450 2D6 metabolism of patients seems to have little effect on safety or tolerability of the drug. This article will review the data from completed and ongoing clinical trials available at the time the New Drug Application was submitted to the FDA. Described are serious adverse events, discontinuations, and treatment-emergent adverse events. Specifically, cardiac effects and effects on weight, height, and metabolism that are related to treatment of ADHD with atomoxetine in children and adolescents are discussed. PMID- 12562063 TI - Development of training and education in rehabilitation technology in Tanzania. PMID- 12562065 TI - Sexuality in persons with limb amputation. AB - Eighty-five (85) persons with limb amputation participated in a self-report survey with the aim of describing their physical and psychological status in their sexual life. The questionnaire was organised to reveal not only the physical aspect of sexual life but also life satisfaction. The results indicated that: 1) 43.5% of the respondents talked about sexual issues with somebody in daily life; 2) 60.0% of the respondents engaged in coitus; and 3) the existence of a partner was shown to be a factor associated with satisfaction with sexual life. PMID- 12562064 TI - Psychological issues in prosthetic and orthotic practice: a 25 year review of psychology in Prosthetics and Orthotics International. AB - In the inaugural edition of Prosthetics and Orthotics International, Fishman identified the psychological sciences as one of six indispensable areas of skill and knowledge in professional prosthetic-orthotic practice. Given the journal's significant role in reporting and developing pertinent research and practice, this review assesses the profile of psychology in prosthetic and orthotic research, as evidenced by the content of Prosthetics and Orthotics International since its inception. A MEDLINE search of the journal's abstracts over a twenty five year period was conducted using the search terms: 'psychology', 'psychosocial', 'quality of life', 'developmental' and 'coping'. Results of this search are summarised under the following headings: (a) body image; (b) coping and adjustment; (c) developmental issues; (d) psychosocial well-being; (e) quality of life; and (f) psychological factors leading to amputation. On the basis of this review, the authors conclude by highlighting a number of key areas where the psychological aspects of prosthetics and orthotics warrant further investigation and dissemination. PMID- 12562066 TI - Comparative study of the strength of the flexor and extensor muscles of the knee through isokinetic evaluation in normal subjects and patients subjected to trans tibial amputation. AB - This study compares the strength of the flexor and extensor muscles of the knee in a group (A) of 25 patients with unilateral trans-tibial amputation, regardless of cause, and a group (B) of 27 normal volunteers. Measured parameters were peak bending moment, total work, maximum power and flexor/extensor relation. The dynamometer used was a Cybex model 6000, set at velocities of 60 degrees/s and 180 degrees/s (4 and 20 repetitions). Exercise type was concentric, both for flexion and extension of the knee joint. Patients were grouped according to stump size, age and time since amputation. These patients were tested with their prosthesis. Mean age in group A was 35.9 +/- 13 years (age range: 12-59 years). Mean age in group B was 34 +/- 8 years (age range: 19-56 years). Comparison according to subject sex was similar. Data analysis between the amputated and the sound limb reveals strength deficit (bigger deficit at 180 degrees/s), which relates to age but not to stump size. When compared to non-amputated subjects in the measured parameters, negative relations both in the amputated side and the non-amputated side were found. The authors' conclusion is that revision of the parameters used until then for rehabilitation of the patients subjected to trans tibial amputation is needed. PMID- 12562067 TI - Comparison of the effects of patellar tendon bearing and total surface bearing sockets on prosthetic fitting and rehabilitation. AB - Patellar tendon bearing (PTB) and total surface bearing (TSB) sockets have been used respectively in the prosthetic treatment of 20 trans-tibial amputees to investigate the effectiveness of both sockets on prosthetic fitting and rehabilitation. Data analysis showed that prostheses with TSB sockets were lighter than the prostheses with PTB sockets and better suspension was obtained from TSB prosthetic socket (p<0.05). It was also found that weight acceptance on the amputated side advanced to a more normal value with TSB prostheses (p<0.05). There was a statistically significant difference between the two socket types in walking and in other ambulation activities except sitting and standing up from a chair, in favour of the TSB socket (p<0.05). Consequently, due to the outcome of this study it can be said that TSB prosthetic sockets can be used effectively in the rehabilitation of trans-tibial amputees. PMID- 12562068 TI - A comparison of traditional prosthetic training versus proprioceptive neuromuscular facilitation resistive gait training with trans-femoral amputees. AB - Reports in the rehabilitation literature suggest that patients with trans-femoral amputation ambulate well after suitable prosthetic treatment. The effects of exercise protocols on function, however, have not been documented in this population. This study was conducted to compare the outcome of traditional and proprioceptive neuromuscular facilitation (PNF) techniques on weight bearing and gait. Fifty unilateral trans-femoral amputees who were attending for their first prosthesis, participated in this study. Amputees were randomly assigned into groups receiving the traditional training or PNF. Traditional treatment was consisted of weight-shifting, balancing, stool-stepping and gait exercises. In the other group the same activities were given by PNF. Amputees were trained 30 minutes daily, for a total of 10 treatments. Pre- and post-training assessment included weight bearing measurements by using two bathroom scales and time distance characteristics of gait from footprints. A statistically significant difference was found in all parameters within the groups due to pre- and post training evaluation data (p<0.05), but more obvious improvement was observed in the group who received PNF (p<0.05). The results of the study suggest that the prosthetic training based on proprioceptive feedback was more effective to improve weight bearing and gait when compared with a traditional programme. PMID- 12562069 TI - Water activity limbs. AB - With advances in technology there is an increasing availability of Water Activity Limbs (WALs) and subsequently a greater number of requests for their provision. This study aims to establish a national consensus for indications, recommended best practice and procedures. The study was conducted with 2 rounds of questionnaires sent to 40 doctors, prosthetists and therapists each. The first questionnaire had a list of possible tasks requiring a WAL and respondents were asked to record their personal rating for prescription of each of the indications. Following analysis of the 91 responses, a list of indications, guidelines and procedures was sent to the same 120 respondents, enquiring whether they agreed or disagreed to each recommendation. The tasks orientated questionnaire showed that more than 50% of respondents considered occupation as an absolute indication. Other indications were some specific water sports. Occasional swimming and beach activity were only considered as possible indications. Showering was not considered an indication. The second questionnaire showed an overwhelming agreement to most of the procedures and indications recommended except social reasons for leisure. The authors present recommendations for prescription of WALs as guidelines and procedures based on the national consensus amongst peers. They also recommend a process for establishing evidence in a speciality where there is very little published evidence to recommend best practice. PMID- 12562070 TI - Development and characterisation of electromechanical muscles for driving trans humeral myoelectric prostheses. AB - Recently, attempts have been made to construct actuators with similar behaviour to natural muscles. However the results have been inadequate for application to practical prostheses. For example, muscle wire, which has too low an efficiency to be powered by batteries and McKibben muscles which require two power supplies, one electric and one pneumatic. Electrochemical muscles are still in the development stage and cannot yet be used for prostheses. In this paper, a new electromechanical actuator is presented, which provides rectilinear movement and linear characteristics. This electromechanical actuator is based on a ball screw and rare earth magnet coreless motors. The system has been characterised and some of the most important results are that it produces a force of 167N while developing a velocity of 7x10(-3)m/s. The force developed is proportional to the current drained. Its efficiency is about 32%, its total mass 0.19kg and it is light and compact enough to be used in practical clinical prosthesis. PMID- 12562071 TI - Health complaints and disabilities in patients supplied with foot orthoses for degenerative foot disorders. AB - This exploratory study attempted to estimate the severity of health complaints and disabilities in patients supplied with foot orthoses for degenerative foot disorders in the Netherlands. Information on the severity and the distribution of the complaints in these patients is important to acquire insight in the problems which these patients experience, and moreover is essential for further research, especially for evaluating effects of patients undergoing foot orthosis treatment. Patients with degenerative foot disorders aged 18 years and over were recruited from nine orthopaedic workshops over a period of three months in 2000. One hundred and twenty-two (122) patients were included in the study. Two approaches were used to obtain data. Firstly, shoe technicians and orthotists inspected patients' feet and legs and interviewed them at their initial visit. Data on gender, age, height, weight, existing and prescribed orthotic devices were recorded on a report form. Secondly, patients were asked to fill in a questionnaire assessing type, location, frequency, intensity and duration of health complaints, and disability. Data indicate that females with degenerative foot disorders and foot orthoses are twice as common as men. Patients supplied with foot orthoses are twice as often overweight compared to subjects in the general population. Besides frequent and protracted pain there are also fatigue complaints, particularly in the feet and lower leg. Duration and frequency of the complaints suggest that these patients suffer from a chronic pain syndrome. In addition, the results indicate that the functional level of these patients is below that of the general population, in particular regarding physical activities. PMID- 12562072 TI - Interface pressure and shear stress changes with amputee weight loss: case studies from two trans-tibial amputee subjects. AB - Interface pressures and shear stresses were measured at monthly intervals on two trans-tibial amputee subjects who lost more than 12% of their body weight over the course of the study. For one subject interface pressures and shear stresses during the weight-acceptance phase of gait decreased over the study interval at all 13 sites monitored, while the other subject experienced increased pressures distally but decreased pressures proximally. Subjects' stumps appeared to atrophy over the study interval, increasing distal end and patellar tendon loading, but not increasing interface shear stresses at other locations. Adding socks at the end of the study did not return interface pressures to first session values at all sites. It is expected that local stump shape changes occurred, causing a non uniform change in interface stress patterns. PMID- 12562073 TI - Knee disarticulation after total-knee replacement. AB - An 89-year-old woman who had a total-knee replacement in the past, underwent a knee disarticulation of the same leg because of an ischaemic foot. Eight (8) months postoperatively the stump is fully weight-bearing and the patient is able to walk safely, using a prosthesis and a walking frame. PMID- 12562074 TI - A test model of hip brace for prevention of dislocation after total hip arthroplasty (Zetton Band). AB - Hip dislocations remain an intractable problem in patients with soft tissue impairment, particularly in those with muscle weakness around the hip, such as those who have undergone revision total hip arthroplasty (THA). At the authors' hospital, postoperative dislocations were observed in 10 of 154 hips between January 1985 and June 1988. Five hips required re-replacement. Conventional measures to prevent or treat post-THA dislocations have been anti-dislocation pants for soft fixation and a cast or abduction-forcing braces for firm fixation. However, the anti-dislocation pants for soft fixation were not as effective as indicated by the above 10 postoperative dislocations. The firm fixation techniques are considered to cause a reduction in muscle strength, causing psychological stress and poor activity of daily living (ADL). The authors devised a soft brace for easy application and prepared its test model to prevent muscle weakening, allow stability of the hip during rotation and avoid restrictions in ADL. This brace was applied to a patient who had 3 dislocations in a short period after being discharged who sustained a postoperative dislocation and achieved good results. PMID- 12562075 TI - Gait analysis in prosthetics: opinions, ideas and conclusions. PMID- 12562076 TI - Lepidolide, a novel seco-ring-A cucurbitane triterpenoid from Russula lepida (Basidiomycetes). AB - A novel compound, lepidolide [structure: see text], was isolated from the fruiting bodies of the Basidiomycete Russula lepida. Its structure was established by spectroscopic and chemical means. PMID- 12562077 TI - New diacylated 2-hydroxythymol derivatives from Melampodium divaricatum. AB - Melampodium divaricatum is a medicinal plant, which occurs in Central America. In a recent paper we reported the occurrence of acylated 2-hydroxy thymol glycosides as main constituents in this plant. This paper deals with the isolation of two new 2,5-dihydroxythymol ester derivatives. The formerly reported sesquiterpene lactone mikanokryptin was not found in our plant material. PMID- 12562078 TI - Iridoid and phenolic glycosides from Wulfenia carinthiaca. AB - Two new phenylpropanoid glycosides (2'-O-acetylplantamajoside and 2'-O, 6"-O diacetylplantamajoside), a new iridoid glycoside (10-O-(cinnamoyl)-6'-O (desacetylalpinosidyl)-catalpol), the two known iridoid glycosides globularin and isoscrophularioside, and the known phenylpropanoid glycoside platamajoside were isolated from the methanolic extract of the underground parts of Wulfenia carinthiaca. Structure elucidations were based on high-resolution mass spectrometry and extensive 1-D and 2-D NMR spectroscopy. PMID- 12562079 TI - Sesquiterpenes and flavonoid aglycones from a Hungarian taxon of the Achillea millefolium group. AB - The investigation of a dichloromethane extract of flower heads of a Hungarian taxon of the Achillea millefolium group led to the isolation of three flavonoid aglycones, one triterpene, one germacranolide and five guaianolides. Their structures were elucidated by UV-VIS, EI- and CI-MS, 1H NMR and 13C NMR spectroscopic methods as well as by 2D-NMR studies and by selective 1D-NOE experiments. Besides apigenin, luteolin and centaureidin, beta-sitosterol, 3beta hydroxy-11alpha,13-dihydro-costunolide, desacetylmatricarin, leucodin, achillin, 8alpha-angeloxy-leucodin and 8alpha-angeloxy-achillin were isolated. Both latter substances are reported here for the first time. Their NMR data were compared with those of the other guaianolides. The stereochemistry of 3beta-hydroxy 11alpha,13-dihydro-costunolide was discussed and compared with data of the literature. PMID- 12562080 TI - Flavonoids and arbutin from Turnera diffusa. AB - The infusion of the aerial parts of Turnera diffusa was phytochemically examined. Chromatographic procedures led to the isolation of a new flavone glycoside, five known flavonoids and p-arbutin. Structures were determined by 1D- and 2D NMR experiments, as well as ES-MS and UV spectra. PMID- 12562082 TI - Chemical composition of volatile oil from Cinnamomum zeylanicum buds. AB - The hydro-distilled volatile oil of the Cinnamomum zeylanicum (C. zeylanicum) buds was analyzed using GC and GC-MS for the first time. Thirty-four compounds representing approximately 98% of the oil was characterized. It consists of terpene hydrocarbons (78%) and oxygenated terpenoids (9%). alpha-Bergamotene (27.38%) and alpha-copaene (23.05%) are found to be the major compounds. A comparison of the chemical composition of the oil was made with that of flowers and fruits. PMID- 12562081 TI - A new phenylpropanoid ester from the bark of Zanthoxylum scandens (Rutaceae). AB - The alkaloids norchelerythrine, magnoflorine and (-)(S)-O-methylbalfourodinium cation were isolated from Zanthoxylum scandens bark collected in Vietnam, together with the flavanone glycoside hesperidin and the phenylpropanoids (E)-O geranylconiferyl alcohol and (E)-O-geranylconiferyl alcohol (9Z, 12Z)-linoleate. This latter is a novel compound whose structure was elucidated on the basis of its spectral data and confirmed by chemical correlation. PMID- 12562083 TI - Sesquiterpene lactones in a hairy root culture of Cichorium intybus. AB - A transformed root culture of Cichorium intybus L. (Asteraceae) was found to produce sesquiterpene lactones of guaiane and germacrane type. Lactucopicrin, 8 desoxylactucin and three sesquiterpene lactone glycosides: crepidiaside B, sonchuside A and ixerisoside D were isolated from the roots. The yield of 8 desoxylactucin reached 0.03 g l(-1) at the early stationary phase of the culture. PMID- 12562084 TI - The chemical composition of Microbiota decussata. AB - From the leaves of Microbiota decussata (Cupressaceae) biflavones: cupressuflavone, amentoflavone and 7-O-methylamentoflavone were isolated and identified. The amount of cupressuflavone in Microbiota decussata and xCupressocyparis leylandii (Leyland cypress) (Cupressaceae) was determined by HPLC (1.82% and 0.83%, respectively). The chemical composition of essential oils from bark and leaves of Microbiota decussata was established by GC-MS (GC-FID) analysis. As a major component thujopsene (39.2% and 45.9%, respectively) was identified. Wiridiflorol (3.0%) and tau-muurolol (0.3%) were present only in leaves but globulol (1.5%) exclusively in bark. The content of essential oils in M. decussata was high--5.4% in bark and 12.6% in leaves. The essential oils from M. decussata and xC. leylandii were bioassayed towards different fungi of the genus Fusarium. Leyland cypress essential oil at 2% concentration fully inhibited the growth of all fungi. PMID- 12562085 TI - 5-(2'-oxoheptadecyl)-resorcinol and 5-(2'-oxononadecyl)-resorcinol, cytotoxic metabolites from a wood-inhabiting basidiomycete. AB - 5-(2'-oxoheptadecyl)-resorcinol [structure: see text] and 5-(2'-oxononadecyl) resorcinol [structure: see text] were isolated from fermentations of an imperfect basidiomycete. The structures of the compounds were determined by spectroscopic techniques. Both compounds exhibit cytotoxic effects against the human colon tumor cell lines COLO-320, DLD-1 and HT-29 and the human promyeloid leukemia cell line HL-60, the human leukemia T cell JURKAT, the human hepatocellular carcinoma cell line HEP-G2 as well as the J774 mouse macrophage cell line. The compounds induce morphological and physiological differentiation of HL-60 cells into granulocytes, which subsequently die by apoptosis. Both compounds show no antibacterial and antifungal activity. PMID- 12562086 TI - Photosynthetic electron transport inhibition by 2-substituted 4-alkyl-6 benzylamino-1,3,5-triazines with thylakoids from wild-type and atrazine-resistant Chenopodium album. AB - The effect of 2-benzylamino-1,3,5-triazines on photosynthetic electron transport (PET) was measured with thylakoids isolated from atrazine-resistant, wild-type Chenopodium album, and spinach to find novel 1,3,5-triazine herbicides bearing a strong PET inhibition. The PET inhibition assay with Chenopodium (wild-type and resistant), yielded a resistance ratio (R/W = I50 (resistant)/I50 (wild-type)) of 324 for atrazine while for benzylamino-1,3,5-triazine derivatives of diamino 1,3,5-triazines a R/W of 11 to 160 was found. The compounds having a benzylamino group at one of the amino groups in the diamino-1,3,5-triazines have a resistant ratio down to one half to 1/30 of the atrazine value. The average resistance ratio of 21 benzylamino derivatives of monoamino-1,3,5-triazines was found to be about 4.0. The inhibition of 21 benzylamino-1,3,5-triazines assayed with atrazine resistant Chenopodium thylakoids, indicated by pI50 (R)-values, correlated well with the PET inhibition pI50 (W) of wild-type thylakoids from Chenopodium. PMID- 12562087 TI - Antibacterial diterpenoids from Astragalus brachystachys. AB - Antibacterial bioassay guided fractionation of acetone extracts of Astragalus brachystachys resulted in isolation of sclareol and two related labdane-type diterpenoids, 14R-epoxysclareol and 6beta-hydroxysclareol. The antibacterial activity of the isolated compounds was measured and it was deduced that the epoxidation at the double bond of sclareol or hydroxylation at C-6 decreased the activity of the resulting compounds. Salvigenin (5-hydroxy-4',6,7 trimethoxyflavone) was also separated from this plant for the first time. PMID- 12562088 TI - Herbal remedies traditionally used against malaria in Ghana: bioassay-guided fractionation of Microglossa pyrifolia (Asteraceae). AB - Different extracts from 11 West African plants traditionally used against malaria in Ghana were tested against both the chloroquine-sensitive strain PoW and the chloroquine-resistant clone Dd2 of Plasmodium falciparum. Due to the promising in vitro activity of the lipophilic extract [IC50: 10.5 microg/ml (PoW); 13.1 microg/ml (Dd2)], Microglossa pyrifolia (Lam.) Kuntze (Asteraceae) was chosen for further phytochemical investigation. From active fractions 13 compounds were isolated; their structures were established on the basis of spectroscopic methods. 1-Acetyl-6E-geranylgeraniol-19-oic acid and sinapyl diangelate represent new natural compounds. The two diterpenes E-phytol [IC50: 8.5 microM (PoW); 11.5 microM (Dd2)], and 6E-geranylgeraniol-19-oic acid [IC50: 12.9 microM (PoW); 15.6 microM (Dd2)] proved to be the most active constituents in our test system. PMID- 12562089 TI - Selective action of acetogenin mitochondrial complex I inhibitors. AB - Five annonaceous acetogenins, rolliniastatin-1 [structure: see text], rolliniastatin-2 [structure: see text], laherradurin [structure: see text], squamocin [structure: see text], annonacin [structure: see text], and rotenone as a reference, differing in their NADH oxidase inhibition activity, have been evaluated for antifeedant, insecticidal, trypanocidal and cytotoxic effects on insect, mammalian and tumor cells. All the test compounds were toxic to Leptinotarsa decemlineata, demonstrated selective cytotoxicity to insect Sf9 cells and a panel of tumor cell lines with the multidrug-resistant SW480 (P glycoprotein+, Pgp+) being the most sensitive one. Compounds [structure: see text] and rotenone had post-ingestive effects against Spodoptera littoralis larvae while [structure: see text] and rotenone were active against Trypanosoma cruzi. Based on their biochemical properties (inhibition of the mitochondrial NADH oxidase activity), the in vivo effects of these compounds on S. littoralis and their cytotoxic effects on Sf9 and tumor cells were more predictable than their effect on T. cruzi and mammalian cells. PMID- 12562090 TI - Continuous decomposition of sporopollenin from pollen of Typha angustifolia L. by acidic methanolysis. AB - Sporopollenin from the pollen of Typha angustifolia L. was exposed to a series of 36 subsequent acidic methanolysis procedures. The remaining decomposition products were investigated using several spectroscopic methods including Fourier transform infrared spectroscopy (FT-IR), solid state 13C nuclear magnetic resonance spectroscopy (13C-CPMAS-NMR) and X-ray photoelectron spectrometry (XPS). Substantial weight losses of the sporopollenin material occur after each acidic methanolysis step, while FT-IR and 13C-CPMAS-NMR spectra display no noticeable differences after 12, 24 and 36 steps. These findings are interpreted as a hint that the sporopollenin polymer has a uniform composition, i.e. relatively small monomer moieties of similar primary structure are present. Moreover, the weight losses account for the presence of substantial amounts of ether linkages in the sporopollenin polymer. PMID- 12562091 TI - Purification and properties of an enzyme capable of degrading the polysaccharide of the cyanobacterium, Nostoc commune. AB - A novel Nostoc commune-polysaccharide (NPS)-degrading enzyme with a molecular mass of 128.5 kDa was purified from Paenibacillus glycanilyticus DS-1. The optimum pH and temperature of the enzyme activity were 5.5 and 35 degrees C, respectively. The enzyme completely degraded NPS to oligosaccharides, ranging from tetra to hexasaccharides and could degrade the xylan weakly whereas xanthan, gellan, cellulose, curdlan and p-nitrophenyl-beta-D-xylopyranoside were not degraded. Homology analysis of the N-terminal amino acid sequence of the NPS degrading enzyme against the PIR and SWISS-PROT databases indicated that the sequence was not homologous to any other polysaccharide-degrading enzyme. PMID- 12562092 TI - Degradation of aqueous 4-chloroaniline by ozonolysis and combined gamma-rays ozone processing. AB - The decomposition of 4-chloroaniline (4-ClA), used as a model for water pollutants, was studied by ozonolysis as well as by gamma-rays in the presence of ozone under comparable conditions. The degradation process was followed by absorption spectroscopy and by HPLC-method as well. Depending on the ozone concentration (mg O3/min) introduced into the aqueous solution the substrate is decomposed to a mixture of carboxylic acids, which can be entirely degradated by prolonged treatment. The combined processing of 4-ClA by gamma-irradiation in the presence of ozone proved to lead even to more efficient degradation of the substrate. Some primary reaction steps are briefly discussed. PMID- 12562094 TI - Isolation and structure elucidation of deformylflustrabromine from the North Sea bryozoan Flustra foliacea. AB - The brominated pyrrolo[2,3-b]indole deformylflustrabromine was isolated as a new natural product from the bryozoan Flustra foliacea, collected in the North Sea. Deformylflustrabromine appears to be the missing link in the biosynthetic sequence from flustrabromine to flustraminol A. Flustramines A, D, and dihydroflustramine C were determined as other major constituents of the investigated sample. Deformylflustrabromine is cytotoxic against the human colon cancer cell line HCT-116 (IC50 5.8 microM). PMID- 12562093 TI - Quick preparative separation of natural naphthopyranones with antioxidant activity by high-speed counter-current chromatography. AB - The natural naphthopyranones paepalantine [structure: see text], paepalantine-9-O beta-D-glucopyranoside [structure: see text] and paepalantine-9-O-beta-D allopyranosyl-(1-->6)-O-beta-D-glucopyranoside [structure: see text] were separated in a preparative scale from the ethanolic extract of the capitula of Paepalanthus bromelioides by high-speed counter-current chromatography (HSCCC). The solvent system used was composed of water-ethanol-ethyl acetate-hexane (10:4:10:4, v/v/v/v). This technique led to the separation of the three different naphthopyranone glycosides in pure form in approximately 7 hours. Paepalantine showed a good antioxidant activity when assayed by the DPPH radical spectrophotometric assay. PMID- 12562095 TI - Adaptation of the phosphotungstate method for the determination of vitamin C contents in animal and human tissues. AB - The usefulness of phosphotungstate reagent for vitamin C determination in tissue homogenates has been confirmed. An optimal homogenization medium was selected: 1.8 M solution of HPO3 in 1.3 M CH3COOH. With this medium the analytical curve (at 700 nm) demonstrated the right linearity, correlation and recovery coefficients were appropriately high (0.999 and 99.8%) and the values of intraserial and interserial variation coefficient were low (< 5% and < 10%, respectively). It makes this method sensitive, easily repeatable, and useful for vitamin C determination in animal and human tissues, including neoplastic ones. PMID- 12562096 TI - Effect of some psychotropic drugs on luminol-dependent chemiluminescence induced by O2-, *OH, HOCl. AB - We studied antioxidant activity of six neuroleptics (chlorpromazine, levomepromazine, promethazine, trifluoperazine and thioridazine) and two antidepressants (imipramine and amitriptyline) in the range of concentration of 10(-7)-10(-4) M. We applied luminol-dependent chemiluminescence to test the ability of these drugs to scavenge the biologically relevant oxygen-derived species: hydroxyl radical, superoxide radical, hypochlorous acid in vitro. We found that the phenothiazines were powerful scavengers of hydroxyl and superoxide radicals. Chlorprothixene, amitriptyline and imipramine had no scavenge activity to the superoxide radical. All drugs showed a moderate scavenger effect on hypochloric anion. PMID- 12562097 TI - Kinetics and mechanism of hydrolysis of acetylthiocholine by butyrylcholine esterase. AB - Kinetics and mechanism of hydrolysis of acetylthiocholine by the enzyme butyrylcholine esterase was studied. The spectrophotometric Ellman's method and potentiometric pH-stat method were used for continuous determination of the actual concentration of the products thiocholine and acetic acid in the reaction mixture. The validity of the Michaelis-Menten (Briggs-Haldane) equation in the whole course of the reaction under used conditions was proved. The corresponding kinetics parameters (Vm and KM) were calculated from the obtained dependences of concentration of thiocholine or acetic acid vs. time and compared. From this comparison the deciding kinetic role of the step producing thiocholine was derived. The values of initial molar concentration of the enzyme and of the rate constants of the kinetic model were estimated. PMID- 12562098 TI - Enzymatic activity and inhibition of the neurotoxic complex vipoxin from the venom of Vipera ammodytes meridionalis. AB - Vipoxin from the venom of Vipera ammodytes meridionalis is an unique neurotoxic complex between a toxic phospholipase A2 and a highly homologous non-toxic protein inhibitor. It is an example of evolution of a catalytic and toxic function into inhibitory and non-toxic one. The activity of the V. ammodytes meridionalis toxin is 1.7 times higher than that of the closely related (92% sequence identity) neurotoxic complex RV4/RV7 from the venom of Vipera russelli formosensis The enhanced enzymatic activity of vipoxin is attributed to limited structural changes, in particular to the substitutions G54R and Q78K in the PLA2 subunit of the complex and to the T54R substitution in the inhibitor. Oleyloxyethylphosphocholine, aristolochic acid and vitamin E suppressed the enzymatic activity of vipoxin and its isolated PLA2 subunit. These compounds influence inflammatory processes in which PLA2 is implicated. The peptide Lys-Ala Ile-Tyr-Ser, which is an integral part of the PLA2 components of the two neurotoxic complexes from V. ammodytes meridionalis and V. russelli formosensis (sequence 70-74) activated vipoxin increasing its PLA2 activity by 23%. This is in contrast to the inhibitory effect of the respective pentapeptides with 70-74 sequences on other group II PLA2s. Surprisingly, the same peptide inhibited 46% of the V. russelli formosensis PLA2 activity. The limited changes in the structure of the two highly homologous neurotoxins lead to considerable differences in their interaction with native peptides. PMID- 12562099 TI - Fluorescence spectroscopy of the tryptophan microenvironment in Carcinus aestuarii hemocyanin. AB - The steady-state and time-resolved fluorescence properties of the multitryptophan minimal subunit CaeSS2 from Carcinus aestuarii hemocyanin have been studied with the aim of probing the environment of the fluorophores within the protein matrix. Subunit a of Panulirus interruptus hemocyanin, whose X-ray structure is known, has been also studied. The results are compared with those collected with other two monomeric fractions (CaeSS1, CaeSS3) produced by dissociation of the native, oligomeric protein as well as with those of the hexameric aggregate. Three classes of tryptophan residues can be singled out by a combination of fluorescence quenching and lifetime measurements on the holo-Hc (the copper containing, oxygen binding form) and the apo-Hc (the copper-free derivative). One class of tryptophans is exposed to the protein surface. Some of these residues are proposed to be involved in the intersubunit interactions in CaeSS1 and CaeSS3 fractions whereas in CaeSS2 the protein matrix masks them. This suggests the occurrence of conformational rearrangements after detachment of the subunit from the native aggregate, which could explain the inability of CaeSS2 to reassociate. A second class of tryptophan has been correlatively assigned, by comparison with the results obtained with Panulirus interruptus hemocyanin, to residues in close proximity to the active site. The third class includes buried, active site distant, residues. PMID- 12562100 TI - Flavonols from Scurrula ferruginea Danser (Loranthaceae). AB - Three natural flavonols compounds have been isolated from the ethyl acetate fraction of Scurrula ferruginea Danser (Loranthaceae). Besides quercetin and quercitrin, an unusual flavonol glycoside 4"-O-acetylquercitrin was isolated. Structures were determined using spectroscopic methods including UV, NMR and HRMS EI. The incidence of 4"-O-acetylquercitrin, not previously reported in the Loranthaceae, is discussed. Cytotoxic evaluation on four human cancer cell lines showed quercetin to be the most active with IC50 of 35 microM on U251 (human glioblastoma cells). PMID- 12562101 TI - Severity of substance abuse and psychiatric problems among perinatal drug dependent women. AB - The present study assessed 108 women enrolled in a residential treatment program for perinatal substance abusers in order to examine possible subtypes based on current alcohol, other drug and psychiatric problem severity. Most participants were African-American (81%), unmarried (91%), unemployed (97%), high school graduates (63%) with a mean age of 30 years. The primary substances of abuse were cocaine/crack (87%) and opiates (9%). The majority (79%) of subjects were also nicotine dependent. Drug severity did not distinguish between these groups; however, when cluster analysis was applied to ASI alcohol and psychiatric composite scores, three clusters emerged: (1) high alcohol, drug, and psychiatric problem severity (57%); (2) high drug and psychiatric problem severity (20%); and (3) high drug severity only (23%). Neither current nor childhood demographic characteristics differed among the three groups. However, significant differences were found among selected childhood risk factors, including perceived quality of relationships with family, age of onset of substance use, childhood abuse history, and family history of mental illness. These results emphasize the need for careful assessment and treatment planning that addresses the unique needs of these women. PMID- 12562102 TI - Sexual abuse among women entering methadone treatment. AB - Women who enter drug abuse treatment programs are likely to report histories of sexual abuse that may impact psychosocial functioning, retention, and outcomes. This study investigates differences at admission between women with and without sexual abuse histories who entered an outpatient methadone treatment program in Texas. In a sample of 137 women, 39% reported prior sexual abuse. Findings show that women with sexual abuse histories were more likely also to have experienced physical and emotional abuse, to report poorer family-of-origin relationships, and to report more drug-related problems. These clients also reported more depression, anxiety, thoughts of suicide, trouble concentrating, and trouble controlling violent behavior. Results are discussed in terms of the need for adequate assessment of abuse history at intake and the need for targeted outcome studies to better define the impact of past abuse on treatment participation and outcome. PMID- 12562103 TI - Treatment concerns of women with co-occurring serious mental illness and substance abuse disorders. AB - The treatment needs of persons with co-occurring mental illness and substance abuse disorders is a relatively new area of inquiry. Even less information is available concerning the treatment concerns of women with dual diagnosis. A focus group was conducted with seven women as part of a larger study of effective treatments for adults with co-occurring disorders. Women responded to questions about what worked and what did not work in their past treatment experiences and what needs to be added for effective treatment. Five primary themes emerged: negative treatment experiences, negative system experiences, desirable treatment characteristics, therapeutic client characteristics, and life issues affecting treatment engagement. Two of the main treatment recommendations that emerged were the need for advocacy assistance with child protective service agencies and the need for providers to use client-centered treatment methods. PMID- 12562104 TI - Drug-scene roles and HIV risk among Puerto Rican injection drug users in East Harlem, New York and Bayamon, Puerto Rico. AB - This article describes and compares distributions of drug-scene roles, frequency of engaging in role behaviors, and relationships of role-holding to high-risk behaviors and sexual partnerships among Puerto Rican injection drug users in New York and Puerto Rico. For this study 561 street-recruited injection drug users in East Harlem, New York, and 312 in Bayamon, Puerto Rico were asked the number of days (in the last 30) in which they earned money or drugs in each of seven drug scene roles; and about behaviors and egocentric risk partner characteristics in the last 30 days. East Harlem subjects were more likely to get resources by selling drugs and syringes, and buying drugs for someone else; Bayamon subjects were more likely to be "hit doctors," buy needles for others, operate a shooting gallery, or escort others to shooting galleries. All roles were part-time except shooting gallery management in East Harlem. About 27% of respondents at each site engaged in two or more roles. Many roles were associated with increased odds of injecting more than twice a day, receptive syringe sharing, distributive syringe sharing, receptive paraphernalia sharing, and having a drug-injecting sex partner. Drug-scene role structures vary between cities. Most roles are part-time pursuits. Role-holders have higher-risk behaviors and sexual partnerships than other drug injectors. Although further research is needed, drug-scene role holders should be targeted for interventions to affect their own risk and their communications with others. PMID- 12562105 TI - Denial in addiction: toward an integrated stage and process model--qualitative findings. AB - Focus groups were conducted at five chemical dependency treatment facilities assessing residents' (N = 65) conceptualizations of denial and the role it may have played in the pathogenesis and progression of their substance use disorders. Two of the authors read verbatim transcripts of the focus group proceedings and independently developed models of denial based on those data. The first reader identified a core set of interpersonal and attributional processes that appeared to explain why many participants did not perceive their substance abuse and associated problems as clearly aberrant and/or chose not to self-identify as alcohol or other drug dependent until late in the course of their chemical dependency. The second reader identified five stages that clients appeared to pass through as they progressed from no awareness of their substance use disorder to full awareness. A second set of readers then independently developed an integrated stage and process model of denial that incorporated elements of the models identified earlier. Though obviously exploratory in nature, these findings suggest that denial is a far more complex and dynamic set of intra- and interpersonal processes than is generally recognized. PMID- 12562106 TI - Looking beyond stereotypes: exploring variations among crack smokers. AB - Since the onset of the crack epidemic in the U.S. in the 1980s, the social and health consequences to the drug's users from their perspective has received limited attention. This exploratory study was designed to explore heterogeneity among crack users. Based on in-depth interviews with 22 active crack users in Atlanta, Georgia, variations among them were explored. Data analysis involved the constant comparison method as part of the grounded theory approach. A typology was derived that describes four subsets of users. The two most salient dimensions of the typology are the level of protective strategies that enable the user to avoid negative consequences and their frequency of crack use. Daily life structure and the extent to which they have access to crack contributed most strongly to use frequency. The typology includes: (1) stable users; (2) tempted users; (3) grappling users; and (4) immersed users. Increased insight into the heterogeneity among crack users is relevant when designing comprehensive prevention and intervention programs. PMID- 12562107 TI - Exploring benzodiazepine use among Houston arrestees. AB - While marijuana and cocaine are the two most prevalent drugs among arrestees, benzodiazepine use has surpassed that of opiates in several jurisdictions across the United States. Despite this proliferation, few scholarly works have focused on benzodiazepine use among individuals under criminal justice supervision. In the present study, the authors used Chi-square statistics and logistic regression to identify significant associations between recent benzodiazepine use (as measured by urinalysis), demographic characteristics, and alcohol and other drug (AOD) use among a sample of 1,572 adult Houston arrestees surveyed through the Arrestee Drug Abuse Monitoring (ADAM) Program in 1999. Compared to nonusers, benzodiazepine-positive arrestees were more likely to be Black, less likely to have a high school diploma, and more likely to be arrested for a drug- or alcohol related offense. Moreover, analyses indicated that recent barbiturate, heroin, PCP, and marijuana use, as measured by urinalysis, were the strongest predictors of recent benzodiazepine use. Policy implications are assessed in light of the current findings. PMID- 12562108 TI - Hearing loss in amphetamine users. AB - Sensory perceptions are modified by amphetamines. Prolonged and heavy use can lead to neuronal damage, neurotransmitter depletion and receptor supersensitivity. Sensorineural deafness had been reported in chronic users of volatile inhalants, alcohol and heroin. There are no reports of hearing problems with amphetamines. Hearing loss was identified in seven amphetamine-dependent inpatients in the detox unit at Al Amal Hospital, Jeddah. Subjects were men aged 18 and over who met DSM IV criteria for substance dependence. The hearing loss was suspected on clinical grounds during the admission interview, and was quite noticeable. Both ears were similarly affected. The symptom mostly appeared three to eight hours after ingestion of the tablets, and did not develop every time the drug was used. Hearing was apparently recovered in every case within four to ten days of cessation of amphetamine use. During the hearing loss, sounds were perceived as less loud and less sharp compared to normal. These cases suggest that chronic amphetamine use may result in reversible hearing loss. The various mechanisms by which this might occur are discussed. The author concludes that these observations should be replicated with a large group of subjects and recommends further investigations to elucidate the nature and site of the damage. PMID- 12562109 TI - Engaging dual diagnosis patients into treatment: the T.E.E.A.C.H. program. AB - This article outlines a model for engaging dually-diagnosed patients into treatment, particularly those patients in the precontemplation and contemplation phases of treatment. The model first focuses on the development of a positive therapeutic relationship with the patient. It then helps patients to realize both the positive and the negative aspects of their substance use. By acknowledging this central conflict and their ambivalence about using, and delivering hope that patients can learn other ways to obtain the benefits of their substance use without the negative consequences, patients can be engaged into treatment and helped to begin the process of change. PMID- 12562110 TI - Amlodipine reduces blood pressure and headache frequency in cocaine-dependent outpatients. AB - Blood pressure and headache frequency were evaluated in normotensive male and female cocaine-dependent patients (N=43) participating in a placebo-controlled, double-blind trial of amlodipine for the treatment of cocaine dependence. Amlodipine produced a significant reduction in both systolic (p=0.04) and diastolic (p=0.01) blood pressures without producing dizziness or faintness. Placebo subjects had about three times the frequency of headaches compared to the amlodipine-treated subjects (p=0.004). The high frequency of headaches reported by cocaine-dependent individuals was significantly reduced by amlodipine and may reflect improved cerebrovascular tone. PMID- 12562111 TI - Can HCV be prevented among injection drug users? AB - The hepatitis-C virus (HCV) spreads rapidly among injection drug users (IDUs) because each act of sharing injection equipment carries a high risk of transmission. IDUs are hard to reach, especially in the early stages of their drug-using careers. IDUs are also poorly organized for self-help. For these three reasons, prevention campaigns that worked against the spread of HIV among gay men will be far less effective against the spread of HCV among IDUs. The best hope for such HCV prevention lies in (1) education of middle and high school youth on parenteral risks, and (2) normalization of needle exchange. PMID- 12562112 TI - The epidemiology of anxiety disorders: prevalence and societal costs. AB - Anxiety disorders are the most prevalent of psychiatric disorders, yet less than 30% of individuals who suffer from anxiety disorders seek treatment. Prevalence of anxiety disorders is difficult to pinpoint since even small changes in diagnostic criteria, interview tools, or study methodology affect results. Analyses of the largest prevalence studies of psychiatric illnesses in the United States find that anxiety disorders afflict 15.7 million people in the United States each year, and 30 million people in the United States at some point in their lives. Currently, the European Study of Epidemiology of Mental Disorders and the World Health Organization World Mental Health 2000 studies are underway. These studies, which share a similar methodology, will facilitate future worldwide comparisons of the prevalence of anxiety disorders. Anxiety disorders impose high individual and social burden, tend to be chronic, and can be as disabling as somatic disorders. Compared with those who have other psychiatric disorders, people with anxiety disorders are high care utilizers who present to general practitioners more frequently than to psychiatric professionals, placing a strain upon the health care system. The economic costs of anxiety disorders include psychiatric, nonpsychiatric, and emergency care; hospitalization; prescription drugs; reduced productivity; absenteeism from work; and suicide. PMID- 12562114 TI - The management of panic disorder. AB - The evidence for benzodiazepines in panic disorder is compelling; along with the selective serotonin reuptake inhibitors (SSRIs), they are a standard treatment for panic and other anxiety disorders. However, extended-release formulations of these agents may prove to be as effective as the immediate-release formulations, and extended-release agents have clinical benefits that may make them more attractive treatments than the currently available, shorter-acting benzodiazepines. Because of their longer duration of action, extended-release benzodiazepines can protect against breakthrough anxiety and need to be taken only once or twice a day, which may improve compliance in some patients. Because the other standard treatments of panic disorder, the SSRIs, have a slow onset of action, adding an extended-release benzodiazepine to the treatment regimen for the initial 6 to 8 weeks could serve as an effective bridge until the desired SSRI effect is realized. PMID- 12562113 TI - Pharmacotherapy of generalized anxiety disorder. AB - Less than one third of people afflicted with generalized anxiety disorder (GAD) experience spontaneous remission, and the symptoms of GAD wax and wane throughout a person's life. The burden of GAD may be reduced with psychopharmacologic therapy. The medications with the most evidence of efficacy in GAD are the benzodiazepines, including a new long-acting formulation of alprazolam. These drugs have a low incidence of side effects but may cause physical dependence, withdrawal, and sedation. Antidepressants are also efficacious in GAD but act less quickly than benzodiazepines. Tricyclic antidepressants such as imipramine may substantially reduce symptoms of anxiety but are not considered a first-line therapy because of their side effects spectrum. The extended-release formulation of venlafaxine and selective serotonin reuptake inhibitors such as paroxetine and sertraline are also efficacious in GAD. While their association with sexual dysfunction may be intolerable for some adults, these drugs may be more appropriate than the benzodiazepines because their chronic use does not lead to dependence. Buspirone also significantly reduces symptoms of GAD and is associated with less sexual dysfunction than SSRIs and less sedation than benzodiazepines. Combining antidepressant and benzodiazepine therapy or medication treatment and psychotherapy may lead to an increase in improvement in patients not responding to 1 treatment approach alone. The most effective treatment for managing the recurrent symptoms of this chronic disorder will remain unknown until more long-term studies using both drug and nondrug therapies are conducted. Remission rates are still only about 40%, signifying the need for improved treatment interventions. PMID- 12562115 TI - Anxiety associated with comorbid depression. AB - Historically, the clinical term for mixed depression and anxiety was anxious depression. With the publication of DSM-III-R, 2 categories were established for the purpose of classifying disorders that involve both anxiety and depression, and that classification system is currently used in DSM-IV as well. These more specific diagnostic criteria have given us a much better understanding of the anxiety spectrum, but have created a need for a better understanding of the place of benzodiazepines in clearly defined indications on the anxiety spectrum. In spite of warnings about side effects, misuse, and dependence, benzodiazepines are frequently prescribed as adjunctive therapy to antidepressants for comorbid anxiety and depression. This article presents data on the prevalence, course, and outcome of comorbid anxiety and depression. It also compares efficacy data from trials of benzodiazepines used alone and in combination with antidepressants for the treatment of anxiety disorders comorbid with depression. PMID- 12562116 TI - The role of extended-release benzodiazepines in the treatment of anxiety: a risk benefit evaluation with a focus on extended-release alprazolam. AB - Immediate-release (IR) benzodiazepines have a short duration of therapeutic effect and are generally less effective for anxiety than selective serotonin reuptake inhibitors in reducing concomitant depressive symptomatology. Common criticisms of benzodiazepines also include the patient's tendency to develop a tolerance to the anxiolytic effect and a dependence on the drug itself. The newer extended-release (XR) benzodiazepine formulation was designed to increase efficacy, duration of therapeutic effect, tolerance, compliance, and ease of discontinuation. The XR benzodiazepine alprazolam has shown efficacy in panic disorder and generalized anxiety disorder comparable to the older benzodiazepine formulations. Pharmacokinetic data show that the XR formulation has a longer therapeutic effect compared with IR formulations, which reduces the potential for breakthrough anxiety symptoms. Data also indicate that the XR formulation has less abuse liability than the IR formulation. This article reviews the efficacy, safety, and discontinuation data from clinical trials of IR and XR benzodiazepines in the treatment of anxiety disorders and provides guidelines to minimize the risk of withdrawal syndrome during benzodiazepine discontinuation. PMID- 12562117 TI - Can color Doppler sonography aid in the prediction of malignancy of thyroid nodules? AB - OBJECTIVE: To determine whether color Doppler interrogation of a thyroid nodule can aid in the prediction of malignancy. METHODS: We obtained color Doppler images of thyroid nodules undergoing sonographically guided fine-needle aspiration. The color Doppler appearance of each nodule was graded from 0 for no visible flow through 4 for extensive internal flow. The size, sonographic appearance, results of fine-needle aspiration, and surgical pathologic findings, if available, were recorded for each nodule. RESULTS: There were 254 nodules sampled, of which 32 were malignant (all confirmed at surgery) and 177 were benign. Fourteen (43.8%) of the 32 malignant nodules were color type 4, compared with only 26 (14.7%) of the 177 benign nodules (P = .0004, Fisher exact test). Thirteen (40.1%) of the 32 malignant nodules were solid, as were 18 (10.2%) of the 177 benign nodules (P = .006, Fisher exact test). Among solid nodules, the prevalence of malignancy was greater when the nodule was hypervascular (13 [41.9%] of 31) than when the color type was less than 4 (11 [14.7%] of 77; P = .004, Fisher exact test). CONCLUSIONS: Solid hypervascular thyroid nodules have a high likelihood of malignancy (nearly 42% in our series). The color characteristics of a thyroid nodule, however, cannot be used to exclude malignancy, because 14% of solid nonhypervascular nodules were malignant. PMID- 12562118 TI - Differential diagnosis of focal epididymal lesions with gray scale sonographic, color Doppler sonographic, and clinical features. AB - OBJECTIVE: To determine whether focal epididymal lesions can be differentiated on gray scale sonographic, color Doppler sonographic, and clinical features. METHODS: This was a retrospectiveanalysis of 60 focal epididymal lesions in 57 patients. Focal epididymal lesions were classified into 3 groups: nonspecific epididymitis (n = 43), tuberculous epididymitis (n = 10), and benign epididymal masses (n = 7). The following gray scale sonographic, color Doppler sonographic, and clinical features were analyzed: size, location, echogenicity, and heterogeneity of the lesion; hypoechoic or hyperechoic rim presence; hydrocele presence; degree of blood flow in the lesion; patient's age; duration of symptoms; and scrotal tenderness. RESULTS: Lesions were larger in patients with tuberculous epididymitis than in those with either nonspecific epididymitis (P = .007) or benign epididymal masses (P = .0017). The hypoechoic or hyperechoic rim of the lesion was more common in patients with benign epididymal masses than in those with nonspecific epididymitis (P = .002). The degree of blood flow in the lesion was greater in patients with nonspecific epididymitis than in those with either tuberculous epididymitis (P = .0019) or benign epididymal masses (P < .001). The duration of symptoms was shorter in patients with nonspecific epididymitis than in those with either tuberculous epididymitis (P < .001) or benign epididymal masses (P = .0092). The frequency of scrotal tenderness was higher in patients with nonspecific epididymitis than in those with either tuberculous epididymitis (P < .001) or benign epididymal masses (P < .001). CONCLUSIONS: Gray scale sonographic, color Doppler sonographic, and some clinical features may be helpful for differential diagnosis of focal epididymal lesions. PMID- 12562119 TI - Power Doppler assessment of vascular changes during fracture treatment with low intensity ultrasound. AB - OBJECTIVE: To study the use of power Doppler sonography for assessing changes in vascularity during treatment of fracture sites with low-intensity ultrasound. METHODS: In 6 dogs (3 pairs), subcutaneous dissection of the midshaft of the ulna allowed symmetrical osteotomies to be made with a reciprocation saw. Three dogs were treated and 3 were used as controls. The fracture site was subjected to 1.5 MHz low-intensity ultrasound (30 mW/cm2) for 20 minutes daily from a therapeutic ultrasonic device. Gray scale sonography was performed to evaluate the fracture site. Power Doppler sonography was used to assess flow patterns at the fracture site and surrounding soft tissue. A sonographic contrast agent was administered intravenously. The dose was 0.2 mL/kg. RESULTS: Power Doppler sonography showed an increase in flow around the fracture site in the treated dogs compared with that in control dogs. Color pixel values on day 7 were 3-fold higher in treated legs than control legs and on day 11 were 33% higher in treated legs than control legs. Enhancement after contrast agent administration was consistently higher in treated legs than control legs. CONCLUSIONS: Power Doppler sonography showed increased vascularity around the fracture sites in treated dogs with and without contrast agent administration. PMID- 12562120 TI - Association of shoulder sonographic findings with subsequent surgical treatment for rotator cuff injury. AB - OBJECTIVE: To identify any clinical and radiologic findings of rotator cuff injury that predict whether patients will undergo shoulder surgery. METHODS: We retrospectively studied all shoulder sonograms obtained at a single institution over 12 months. Possible predictors of surgical treatment were documented, including patient age and sex, duration and types of symptoms, and the location and severity of tendon damage on sonography. One hundred twenty-eight patients underwent shoulder sonography; 34 patients eventually underwent shoulder surgery. Multivariate logistic regression was performed to identify clinical and sonographic factors associated with the use of surgical therapy. RESULTS: The only statistically significant predictor of surgical intervention was the finding of full-thickness tears (with or without tendon retraction) on sonography (P = .03). Patients with full-thickness tears were 4.3 times more likely to undergo surgery than those with no tears (odds ratio, 4.3). CONCLUSIONS: The sonographic diagnosis of full-thickness rotator cuff tears is the only finding statistically associated with the use of surgical treatment. No single clinical variable was consistently associated with subsequent surgery. Partial-thickness tears on sonography also do not show any statistical association with the eventual use of surgery for rotator cuff symptoms. PMID- 12562121 TI - The ultrasonographic appearance of the femoral nerve and cases of iatrogenic impairment. AB - OBJECTIVE: To assess the feasibility of ultrasonography of femoral nerves in a cadaveric specimen, healthy volunteers, and patients. METHODS: In 1 unembalmed cadaveric specimen (female, 90 years) and 20 healthy volunteers (9 male and 11 female, 18-50 years; n = 40 scans), the topographic features, cross-sectional shapes (oval or triangular), and cross-sectional areas of the femoral nerves were evaluated by ultrasonography (5- to 12-MHz broadband linear array). In a subsequent study, 7 consecutive patients with postoperative findings assigned to the femoral nerve were evaluated and assessed by a neurologist. RESULTS: The mean +/- SD anteroposterior and mediolateral diameters of the femoral nerves in the volunteers were 3.1 +/- 0.8 and 9.8 +/- 2.1 mm, respectively, at an average cross sectional area of 21.7 +/- 5.2 mm2. The cross-sectional shape was oval in 67.5% superior to the inguinal ligament and in 95% inferior to the ligament. The infrainguinal femoral nerve showed variable distances to the femoral artery. In the subsequent patient study, 5 patients had swelling of the femoral nerve in the affected side. In 1 patient, the nerve had a blurred echo structure due to a hematoma. In 1 patient, major damage of the femoral nerve was ruled out clearly. CONCLUSIONS: Ultrasonography allows the depiction and assessment of the femoral nerve from about 10 cm superior to 5 cm inferior to the inguinal ligament. In this region, ultrasonography is helpful in detection of impairments and, therefore, in decisions about planning and even acceleration of further treatment. PMID- 12562122 TI - Transthoracic sonography of diffuse parenchymal lung disease: the role of comet tail artifacts. AB - OBJECTIVE: The value of transthoracic sonography in diffuse parenchymal lung disease is not established. This prospective study was conducted to analyze pleural and parenchymal alterations in patients with diffuse parenchymal lung disease by transthoracic sonography and to compare the results with the findings of a control group. METHODS: Fifty-three patients with diffuse parenchymal lung disease and 35 control subjects underwent transthoracic sonography for assessment of (1) basal pleural effusion, (2) the presence and number of comet tail artifacts, (3) thickening, (4) an irregular, fragmented pleural line, and (5) subpleural alterations. RESULTS: Basal pleural effusion was revealed in 37.7% of the patients with diffuse parenchymal lung disease (0% of the control subjects; P < .001), and 98.1% had multiple (>6 per scan) comet tail artifacts (14.3% of the control subjects; P < .001). Only a few artifacts (< or = 6 per scan; P < .001) were shown in 85.7% of the control subjects. All the control subjects had strongly localized comet tail artifacts and no thickened pleural line (84.9% of patients; P < .001). In 98.1 % of the patients, an irregular, fragmented pleural surface could be detected (82.8% of the control subjects; P < .05). Subpleural alterations were found in 37.7% of the patients and in 8.6% of the control subjects (P < .001). CONCLUSIONS: Diffuse parenchymal lung disease should be considered if multiple comet tail artifacts distributed over the whole surface of the lung together with a thickened and irregular, fragmented pleural line are visible. Transthoracic sonography may reflect the distribution of pleural involvement and may show subpleural alterations. PMID- 12562123 TI - Comparison of three- and two-dimensional sonography in diagnosis of gallbladder diseases: preliminary experience. AB - OBJECTIVE: To compare three-dimensional sonography with two-dimensional sonography for diagnosis of gallbladder diseases. METHODS: Sixty-three patients with gallbladder diseases were examined with two- and three-dimensional sonography. The morphologic features presented on both modalities were analyzed, and the diagnostic accuracies of both modalities were evaluated and compared. RESULTS: Both two- and three-dimensional sonography facilitated correct diagnosis in all patients with gallstones. Three-dimensional sonography had no influence on the diagnosis of gallstones compared with two-dimensional sonography. Three dimensional sonography showed the granular surfaces in 18 (81.8%) of 22 cases of polypoid lesions and the pedunculated fundus in 19 (86.4%) of 22, whereas two dimensional sonography displayed them in 10 (45.5%) and 3 (13.6%) of 22, respectively (P < .05 and .001). Three- and two-dimensional sonography made correct differential diagnoses between non-neoplastic and neoplastic polyps in 20 (90.9%) and 12 (54.5%) of 22, respectively (P < .05). In gallbladder carcinoma, both three- and two-dimensional sonography accurately showed all the associated gallstones and intrahepatic bile duct dilatation. Two-dimensional sonography could not correctly define the locations of the lesions in 2 (28.6%) and tumor extension in 1 (14.3%) of 7 cases with pathologically proved gallbladder cancer, whereas three-dimensional sonography improved the diagnosis in these cases. CONCLUSIONS: Three-dimensional sonography adds no advantages for diagnosis of gallstones compared with two-dimensional sonography, but it is better than two dimensional sonography for differential diagnosis of gallbladder polyps and may improve the localization and staging for gallbladder carcinoma; however, additional studies are needed for further determination. PMID- 12562124 TI - Complications after interventional sonography of focal liver lesions: a 22-year single-center experience. AB - OBJECTIVE: To analyze the complications of diagnostic and therapeutic sonographically guided interventional procedures of focal liver lesions observed during a 22-year period in a single center. METHODS: Complications of sonographically guided diagnostic and therapeutic procedures on focal liver lesions, observed during a 22-year period in a single center, were reviewed. From 1979 to 2001, 13,222 patients (age range, 7-89 years; mean, 59 years; 8,688 male and 4,534 female) with 13,777 focal liver lesions underwent 16,648 sonographically guided biopsies and 3,035 therapeutic procedures: pyogenic and amebic abscess aspiration, ethanol injection of hydatid liver cysts, and percutaneous ablative treatments (ethanol injection in either multiple or one shot sessions, radio frequency ablation, and interstitial laser photocoagulation) of primary and secondary liver tumors. RESULTS: The overall mortality was 0.06%. No death or major complication occurred after diagnostic procedures and liver abscess drainage. In the therapeutic group mortality was 0.6%: 1 patient died of anaphylactic shock during treatment of a hydatid cyst; 7 patients died after liver tumor ablation with ethanol injection (6 after one-shot treatments and 1 after multisession treatments). Major complications after liver tumor ablative procedures included 10 cases of acute liver failure, 2 cases of acute tubular necrosis, 2 cases of self-limiting hemoperitoneum, 2 cases of paralytic ileum, 2 abscesses, and 1 case of cholangitis. One case of a biliary cyst fistula and 1 case of intracystic hemorrhage occurred after treatment of hydatid liver cysts. CONCLUSIONS: Sonographically guided diagnostic biopsy of focal liver lesions and liver abscess drainage are safe procedures. In contrast, liver tumor ablation procedures have a low but definite risk of mortality and major complications. Puncture of hydatid cysts must be performed only in institutions that can treat anaphylactic shock. PMID- 12562125 TI - Interlaboratory acoustic power measurement. AB - OBJECTIVE: This article describes an American Institute of Ultrasound in Medicine sponsored intercomparison of the results of acoustic power measurements performed by several laboratories. METHODS: Two primary calibration techniques, namely, planar scanning and radiation force balance, were used in the frequency range typical of that in which sonographic imaging devices operate. The same reference source, the National Institute of Standards and Technology (Gaithersburg, MD) standard ultrasonic power source, capable of producing acoustic fields in the frequency range from approximately 1 to 21 MHz, was circulated to 3 laboratories. RESULTS: The results of the calibrations indicate that the overall uncertainty in acoustic power measurements depends on the target and the measurement method. In the case of radiation force balance measurements with an absorbing target, the largest discrepancy between the available National Institute of Standards and Technology-calibrated results and the reported data was 10.6% at approximately 2.5 MHz. At higher frequencies, beyond 10 MHz, the largest discrepancy reported with an absorbing target was 8.4%. For a reflecting target, the largest discrepancies were 16.2% at approximately 3.7 MHz and 15.4% at about 10 MHz. The largest discrepancy identified for the planar scanning technique below 10 MHz was 7.4% at 3.7 MHz. CONCLUSIONS: The results obtained suggest that an absorbing target may be preferable for acoustic power measurements with radiation force balance. In a group that consists of 2 research laboratories and 1 manufacturer, the power measurements agreed within 16%. PMID- 12562126 TI - Value of four-dimensional ultrasonography in ultrasonographically guided biopsy of hepatic masses. AB - OBJECTIVE: To assess the value of four-dimensional ultrasonography (dynamic three dimensional ultrasonography with the added dimension of time) in ultrasonographically guided biopsy procedures of focal hepatic masses. METHODS: Four-dimensional ultrasonographically guided biopsy experiments using a freehand technique were performed in phantoms that simulated human liver tissue and had target material. After the protocol was determined and Institutional Review Board approval was obtained, 12 patients underwent four-dimensional ultrasonographically guided biopsy of focal hepatic masses by informed consent. Planar images using 3 orthogonal planes plus a volume-rendered image were used for real-time guidance during the biopsy procedure. RESULTS: Four-dimensional ultrasonography improved visualization of biopsy devices in all 12 patients (100%) regardless of minor changes in transducer position during the biopsy procedures. Four-dimensional ultrasonography allowed more intuitive apprehension of the spatial relationship of the needle and the target lesion and thus helped in adjusting the needle to an optimal prefiring position in 8 patients (67%). With regard to procedure time, four-dimensional ultrasonographically guided biopsy was comparable with standard two-dimensional ultrasonographically guided techniques. CONCLUSIONS: Compared with standard two-dimensional ultrasonographically guided biopsy, four-dimensional ultrasonography provides improved visualization of biopsy devices and more perceptible information on the spatial relationship between the biopsy needle and the target lesion. PMID- 12562127 TI - Gonococcal arthritis of the shoulder and septic extensor tenosynovitis of the wrist: sonographic appearances. PMID- 12562128 TI - Renal agenesis and seminal vesicle cyst. PMID- 12562129 TI - Twinkling artifact in gallbladder adenomyomatosis. PMID- 12562130 TI - Valvular-type membranous obstruction of the inferior vena cava with vertebral collaterals: color Doppler findings. PMID- 12562131 TI - Private matters. PMID- 12562132 TI - Adopted children: core issues and unique challenges. AB - TOPIC: Psychological impact of adoption and resulting core issues for adopted children. PURPOSE: To raise awareness of the psychological impact of adoption. SOURCES: Published literature and personal observations. CONCLUSIONS: Adoption raises unique issues and challenges for the child and adoptive parents. Resolving the issues of adoption is a lifelong process. Through an awareness of the issues inherent in adoption, nurses and parents can use strategies that will enhance children's self-esteem and decrease their emotional vulnerability. Search terms: Adoption, intervention, loss, self-esteem, strategies PMID- 12562133 TI - Caregiver understanding of adolescent development in residential treatment. AB - PROBLEM: Caretaker knowledge and understanding of adolescent development and its application to clinical practice with severely emotionally disturbed adolescents in residential treatment. METHODS: Twenty child-care workers and registered nurses participated in semistructured interviews analyzed using dimensional analysis, a grounded theory method. FINDINGS: Three distinct categories of caregivers were identified based on level of expertise and engagement in developmentally appropriate treatment practices: inexperienced, party-liners, and transcenders. Developmental issues identified included lack of resident preparation for puberty and staff discomfort with adolescent sexuality. CONCLUSIONS: Caregiver, institutional, and social barriers to developmentally sensitive practice were identified. Practice recommendations include direct preparation of children and adolescents in residential treatment for pubertal changes and sexual development, and careful discernment of age-appropriate and psychopathological adolescent behaviors. PMID- 12562135 TI - Cardiac conduction and antipsychotic medication: a primer on electrocardiograms. PMID- 12562134 TI - Survival against drugs: education for school-age children. AB - PROBLEM: Alcohol and drug use of young school-age children continue to escalate. Comprehensive, effective interventions are needed to treat and prevent future alcohol and drug use. METHODS: The alcohol and drug use of 69 school-age children participating in afterschool programs was explored; parents completed a family climate scale. An investigator-developed educational program was evaluated for its effect on self-efficacy of the children to prevent drug and alcohol use. FINDINGS: Although family climate scales indicated functioning families, 25% of the children indicated they have used drugs or alcohol and 49% indicated a possible problem with alcohol or drugs being used at home by parents. A significant correlation with a child's self-efficacy and drug use was found. CONCLUSIONS: The educational program taught children survival skills to resist the use of alcohol and drugs. Children with a plan to resist the use of drugs were more likely not to use drugs. PMID- 12562136 TI - On ADHD and smoking. PMID- 12562137 TI - Neurobiology of substance P and the NK1 receptor. AB - Substance P belongs to a group of neurokinins (NKs), small peptides that are broadly distributed in the central nervous system (CNS) and peripheral nervous system (PNS). The biological effects of substance P in the CNS, namely regulation of affective behavior and emesis in the brain and nociception in the spinal cord, are mediated by its binding to the NK1 receptor. The substance P-NK1 (SP-NK1) receptor system is the most extensively studied NK pathway, and in contrast to receptors for other neurotransmitters, such as glutamate, which have high expression throughout the CNS, only a minority of neurons (5% to 7%) in certain CNS areas express the NK1 receptor. The NK1 receptor is distributed in the plasma membrane of cell bodies and dendrites of unstimulated neurons, but upon substance P binding, the NK1 receptor undergoes rapid internalization, followed by rapid recycling to the plasma membrane. Release of substance P is induced by stressful stimuli, and the magnitude of its release is proportional to the intensity and frequency of stimulation. More potent and more frequent stimuli allow diffusion of substance P farther from the site of release, allowing activation of an approximately 3- to 5-times greater number of NK1 receptor-expressing neurons. Recent studies employing pharmacologic or genetic inactivation of NK1 receptors demonstrate the important role of the SP-NK1 receptor system in the regulation of affective behavior and suggest that inhibition of this pathway may be a useful approach to treatment of depression and associated anxiety. PMID- 12562138 TI - Behavioral and physiologic effects of genetic or pharmacologic inactivation of the substance P receptor (NK1). AB - Depression and anxiety are among the most common diseases in the United States, thus constituting a substantial financial burden for the health care system. Experimental studies of these affective disorders to date have largely focused on the neurotransmitter pathways with well-established pathophysiologic roles, such as serotonergic, noradrenergic, and gamma-aminobutyric acid (GABA)-ergic systems; agents modulating the activity of these pathways are known to be clinically effective. More recently, the neuropeptide substance P (SP) and its receptor (the neurokinin-1 receptor [NK1R]) have been implicated in the pathophysiology of affective disorders, including depression. Earlier preclinical and clinical studies, though, did not provide a clear consensus on the role of SP in the regulation of affective behavior and related pathologic conditions. Recent studies in mice clearly demonstrate that both the genetic disruption and acute pharmacologic blockade of the NK1R result in marked reduction in anxiety-like behavior and stress-related responses. In parallel with these behavioral effects, physiologic changes, such as an increased firing rate of 5-hydroxytryptamine (5 HT) neurons in the dorsal raphe nuclei and a desensitization of presynaptic 5 HT1A inhibitory autoreceptors, were observed. These findings provide further evidence for the regulatory role of the SP-NK1R system in modulation of affective behavior and indicate that its effects are mediated, at least in part, via the serotonergic system. Future studies will attempt to delineate the interaction between the SP-NK1R system and various neurotransmitter pathways in greater detail and to address the specific role(s) of this system in different brain regions. PMID- 12562139 TI - Imaging substance P receptors (NK1) in the living human brain using positron emission tomography. AB - Substance P (SP)-neurokinin-1 (NK1) receptor pathways have been implicated in the pathophysiology of emesis and depression. Autoradiographic studies in monkey and human brains have shown a high expression of NK1 receptors in regions important for the regulation of affective behaviors and the neurochemical response to stress. Furthermore, clinical studies demonstrated that treatment with the SP (NK1 receptor) antagonist (SPA) aprepitant (also known as MK-0869) significantly improves depression symptoms and reduces the incidence of chemotherapy-induced nausea and vomiting. An important objective of all neuroscience drug discovery and development programs is to establish the correlation between dose, receptor occupancy, and the observed clinical effect (the dose-response relationship). These goals can be achieved using radioactive receptor-specific tracers and dynamic noninvasive brain imaging modalities, such as positron emission tomography (PET). In the SPA program, a tracer [18F]SPA-RQ was chosen for PET studies on the basis of several criteria, including high affinity for the NK1 receptor, low nonspecific binding, and good blood-brain barrier penetration. PET imaging studies in rhesus monkeys and humans confirmed these tracer features and established the usefulness of this probe for in vivo NK1 receptor occupancy studies. Subsequent PET occupancy studies in humans predicted that very high levels of central NK1 receptor occupancy (> 90%) were associated with therapeutically significant antidepressant and antiemetic effects. Future PET imaging studies will focus on quantification of NK1 receptor expression in depressed patients, both before and after successful treatment with antidepressants. PMID- 12562140 TI - Clinical experience with substance P receptor (NK1) antagonists in depression. AB - Substance P (SP) belongs to the neurokinin (NK) family of neuropeptides and exerts its biological effects via interaction with the NK1 receptor. The SP-NK1 receptor system is one of the best-characterized neurotransmitter pathways in both the central and peripheral nervous systems. It has been postulated that this pathway may have important roles in a variety of centrally regulated pathophysiologic conditions, including depression. In animal models, central injection of SP was associated with a series of anxiety-like behaviors, and this response could be abolished by pretreatment with SP (NK1) receptor antagonists (SPAs). On the basis of these and other encouraging preclinical results, several clinical trials have examined the potential of SPAs in the treatment of depression. In phase 2 trials, therapy with the SPAs aprepitant (MK-0869) and compound A resulted in improvements in depression and anxiety symptoms that were quantitatively comparable with those seen with selective serotonin reuptake inhibitors (SSRIs) and significantly greater than those seen with placebo. These positive results have established a proof of concept that the inhibition of the SP-NK1 receptor pathway may be a potentially useful novel treatment option for management of patients with depression. The apparent lack of benefit with SPAs versus placebo in subsequent dose-finding studies with aprepitant and compound A is not surprising, considering the fact that the outcomes with an active control (SSRI) in these trials were also similar to those observed with placebo. Future trials with SPAs will focus on the identification of appropriate patients and drug regimens and will also define the role of these agents in the treatment of depression. PMID- 12562141 TI - Quetiapine: preclinical studies, pharmacokinetics, drug interactions, and dosing. AB - Quetiapine is a novel dibenzothiazepine atypical antipsychotic. Quetiapine shows affinity for various neurotransmitter receptors including serotonin, dopamine, histamine, and adrenergic receptors and has binding characteristics at the dopamine-2 receptor similar to those of clozapine. In animal models, the drug has a preclinical profile suggestive of antipsychotic activity with a reduced tendency to cause extrapyramidal symptoms (EPS) and sustained prolactin elevation. For example, quetiapine alters neurotensin neurotransmission and c-fos expression in limbic but not motor brain regions. The drug also demonstrates clozapine-like activity in a range of behavioral and biochemical tests and may possess neuroprotective properties. In humans, quetiapine exhibits linear pharmacokinetics with a mean terminal half-life of 7 hours. The primary route of elimination of quetiapine is through hepatic metabolism. Although not affected by smoking, alterations in quetiapine disposition due to age or hepatic impairment are manageable by appropriate dosage reduction. The optimal dosing range for quetiapine is 150 to 750 mg/day, and recent results suggest that once-daily dosing may be suitable for some patients. Finally, imaging studies with positron emission tomography confirm significant differences between quetiapine and typical antipsychotics that may be indicative of their differences in mechanism of action and propensity for producing EPS. PMID- 12562142 TI - Efficacy, safety, and tolerability of quetiapine in patients with schizophrenia. AB - Schizophrenia is a serious and disabling psychiatric disorder affecting approximately 1% of the world's population, with its economic cost in the United States alone estimated to exceed that of all cancers combined. The new generation of atypical antipsychotics introduced over the past decade have comparable or greater efficacy than traditional antipsychotics in treating the psychotic symptoms of schizophrenia and a much improved neurologic side effect profile. Quetiapine, the fourth atypical antipsychotic marketed in the United States, was approved by the U.S. Food and Drug Administration in September 1997 and is also currently approved in over 70 countries worldwide for the treatment of psychosis associated with schizophrenia. This article will review the clinical trials examining the efficacy, safety, and tolerability of quetiapine in the treatment of patients with schizophrenia. PMID- 12562143 TI - Use of quetiapine in elderly patients. AB - Behavioral problems associated with psychosis in the elderly have a significant negative impact on patients' quality of life and can lead to placement in a nursing home. Because of their decreased propensity to produce extrapyramidal symptoms, atypical antipsychotics such as quetiapine hold promise in the treatment of these vulnerable patients. Quetiapine may, in theory, be particularly advantageous in this regard because of its lack of anticholinergic activity and its relatively loose binding to dopamine receptors. This article reviews the somewhat limited number of clinical studies of the use of quetiapine in treating older patients with schizophrenia and other psychotic disorders, patients with psychosis associated with Alzheimer's disease or dementia with Lewy bodies, and patients with Parkinson's disease and drug-induced psychosis. PMID- 12562144 TI - Use of quetiapine in children and adolescents. AB - The atypical antipsychotic quetiapine has been examined in children and adolescents in a randomized clinical trial, a number of open-label studies, and several chart review studies. Although only a small amount of information exists, most studies indicate that quetiapine is effective and well tolerated in various pediatric populations. Because quetiapine appears to be well tolerated in the young and associated with manifest salutary effects, it seems to be a promising agent that has potential for use in children and adolescents. This article reviews studies of quetiapine in the treatment of children and adolescents with a variety of psychiatric disorders. Despite these encouraging findings, the number of studies is small, and some have methodological limitations. Methodologically rigorous studies with substantive numbers of subjects are needed to confirm or refute these preliminary impressions. PMID- 12562145 TI - Clinical use of quetiapine in disease states other than schizophrenia. AB - Although quetiapine was introduced as an atypical antipsychotic drug with clinical efficacy in schizophrenia patients, it has been used in a variety of disease states over the last 5 years. The most common conditions have included mood and anxiety disorders, obsessive-compulsive disorder, aggression, hostility, posttraumatic stress disorder, borderline personality disorder, delirium, and comorbid substance abuse. Considering its efficacy in a wide variety of neuropsychiatric conditions and its excellent tolerability profile, quetiapine could emerge as a broad-spectrum psychotropic medication that may be helpful in psychiatry across various diagnostic categories. Traditionally, studies on the predictive validity of psychiatric disorders help with nosologic issues and controversies. Assessing quetiapine's tolerability and its overall treatment response might help tease out the predictive validity of various psychiatric syndromes (based currently on an atheoretical descriptive approach) and may shape psychiatric nosology in the future. Quetiapine's low affinity and fast dissociation from postsynaptic dopamine-2 receptors give the least risk of producing acute extrapyramidal side effects, tardive dyskinesia, and neuroleptic malignant syndrome. These factors suggest that the clinical utility of quetiapine in psychiatric conditions other than schizophrenia has not been fully exploited thus far. PMID- 12562146 TI - Prevention and control of infections with hepatitis viruses in correctional settings. Centers for Disease Control and Prevention. AB - This report consolidates previous recommendations and adds new ones for preventing and controlling infections with hepatitis viruses in correctional settings. These recommendations provide guidelines for juvenile and adult correctional systems regarding 1) identification and investigation of acute viral hepatitis; 2) preexposure and postexposure immunization for hepatitis A and hepatitis B; 3) prevention of hepatitis C virus infection and its consequences; 4) health education; and 5) release planning. Implementation of these recommendations can reduce transmission of infections with hepatitis viruses among adults at risk in both correctional facilities and the outside community. These recommendations were developed after consultation with other federal agencies and specialists in the fields of corrections, correctional health care, and public health at a meeting in Atlanta, March 5-7, 2001. This report can serve as a resource for those involved in planning and implementing health-care programs for incarcerated persons. PMID- 12562147 TI - Differential diagnosis for chronic fatigue syndrome. PMID- 12562148 TI - Using a metal detector to track a swallowed penny. PMID- 12562149 TI - Erythrasma and common bacterial skin infections. PMID- 12562150 TI - Novel cardiovascular risk factors: do they add value to your practice? PMID- 12562151 TI - Diagnosis of eating disorders in primary care. AB - Eating disorders, particularly anorexia nervosa and bulimia nervosa, are significant causes of morbidity and mortality among adolescent females and young women. Eating disorders are associated with devastating medical and psychologic consequences, including death, osteoporosis, growth delay, and developmental delay. Prompt diagnosis is linked to better outcomes. A good medical history is the most powerful tool. Simple screening questions, such as "Do you think you should be dieting?" can be integrated into routine visits. Physical findings such as low body mass index, amenorrhea, bradycardia, gastrointestinal disturbances, skin changes, and changes in dentition can help detect eating disorders. Laboratory studies can help diagnose these conditions and exclude underlying medical conditions. The family physician can play an important role in diagnosing these illnesses and can coordinate the multidisciplinary team of psychiatrists, nutritionists, and other professionals to successfully treat patients with eating disorders. PMID- 12562152 TI - Information from your family doctor. How do I know if I have an eating disorder? PMID- 12562153 TI - Acute respiratory distress syndrome. AB - Acute respiratory distress syndrome is a manifestation of acute injury to the lung, commonly resulting from sepsis, trauma, and severe pulmonary infections. Clinically, it is characterized by dyspnea, profound hypoxemia, decreased lung compliance, and diffuse bilateral infiltrates on chest radiography. Provision of supplemental oxygen, lung rest, and supportive care are the fundamentals of therapy. The management of acute respiratory distress syndrome frequently requires endotracheal intubation and mechanical ventilation. A low tidal volume and low plateau pressure ventilator strategy is recommended to avoid ventilator induced injury. Timely correction of the inciting clinical condition is essential for preventing further injury. Various medications directed at key stages of the pathophysiology have not been as clinically efficacious as the preceding experimental trials indicated. Complications such as pneumothorax, effusions, and focal pneumonia should be identified and promptly treated. In refractory cases, advanced ventilator and novel techniques should be considered, preferably in the setting of clinical trials. During the past decade, mortality has declined from more than 50 percent to about 32 to 45 percent. Death usually results from multisystem organ failure rather than respiratory failure alone. PMID- 12562154 TI - Seizure disorders in the elderly. AB - Seizure disorders become increasingly common after the age of 60 years and can have a significant impact on functional status. The goal of antiepileptic drug therapy is to control seizures but preserve quality of life. If possible, seizure control should be achieved with one agent given in the lowest effective dosage. Clinical response, rather than drug levels, should guide dosage changes. All antiepileptic drugs can cause dose-dependent sedation and cognitive impairment. Although the newer agents may have theoretical advantages over standard antiepileptic agents, higher cost may limit their use. Drugs for first-line monotherapy of seizures in elderly patients include carbamazepine, valproic acid, oxcarbazepine, gabapentin, and lamotrigine. PMID- 12562155 TI - Alternative therapies for traditional disease states: osteoarthritis. AB - Americans spend more on natural remedies for osteoarthritis than for any other medical condition. In treating osteoarthritis, glucosamine and chondroitin sulfate, two of the molecular building blocks found in articular cartilage, are the most commonly used alternative supplements. In randomized trials of variable quality, these compounds show efficacy in reducing symptoms, but neither has been shown to arrest progression of the disease or regenerate damaged cartilage. Although few clinical trials on S-adenosylmethionine exist, preliminary evidence indicates that it relieves pain to a degree similar to that of nonsteroidal anti inflammatory drugs but with fewer side effects. Clinical trials of dimethyl sulfoxide offer conflicting results. Neither ginger nor cetyl myristoleate has proven clinical usefulness. PMID- 12562156 TI - Chronic illness and sexual functioning. AB - Chronic illness and its treatments can have a negative impact on sexual functioning. The mechanism of interference may be neurologic, vascular, endocrinologic, musculoskeletal, or psychologic. Patients may mistakenly perceive a medical prohibition to the resumption of sexual activity, or they may need advice on changes in sexual activity to allow satisfactory sexual functioning. Family physicians must be proactive in diagnosing and managing the alterations in sexual functioning that can occur with chronic illness. Patient education and reassurance are essential. Before sexual activity is resumed, patients with cardiovascular disease should be stratified according to risk. Patients with musculoskeletal disease should be educated about positional changes that may improve comfort during sexual activity. Psychosocial concerns should be addressed in patients with human immunodeficiency virus infection or acquired immunodeficiency syndrome. In patients with cancer, it is important to discuss sexual problems that may arise because of negative body image and the effects of chemotherapy. Patients who have disabilities can benefit from the use of muscle relaxants, technical adaptations, and expansion of their sexual repertoire. PMID- 12562157 TI - Information from your family doctor. Health problems and your sex life. PMID- 12562158 TI - Postmenopausal hormone replacement therapy for the primary prevention of chronic condition. Recommendations and rationale. U.S. Preventive Services Task Force. PMID- 12562159 TI - Information from your family doctor. Exercise: how to get started. PMID- 12562160 TI - Information from your family doctor. Weight-training and weight-lifting safety. PMID- 12562161 TI - Information from your family doctor. Running: avoiding running injuries. PMID- 12562162 TI - NIH releases statement on managing pain, depression, and fatigue in cancer. PMID- 12562163 TI - The use of vibration as an exercise intervention. AB - The use of vibration as a means for enhancing athletic performance is a recent issue in exercise physiology. Current evidence suggests that vibration is effective in enhancing strength and the power capacity of humans, although the mechanisms mediating this effect are unknown. PMID- 12562164 TI - Role of body size in the relation between muscle strength and movement performance. AB - Body size relates differently on the outcome of various tests of muscle strength and movement performance. Thus the estimation of functional performance from tests of muscle strength, and vice versa, should be based on the normalization of both muscle strength and movement performance. PMID- 12562165 TI - Adrenergic contribution during acclimatization to high altitude: perspectives from Pikes Peak. AB - We have examined the sympathoadrenal responses to both acute and chronic high altitude exposure at the summit of Pikes Peak, CO, in both men and women. A dissociation between the adrenal medullary response (acute) with that of the sympathetic nervous system (chronic) is observed. Both alpha- and beta-adrenergic contributions to key metabolic and physiologic adjustments to high-altitude exposure are evident. PMID- 12562166 TI - Movement termination and stability in standing. AB - This brief review develops a new conceptual framework for describing the influence of center-of-mass position and velocity on ability to terminate movement. Limitations of this dynamic conceptual framework and its applications in physical rehabilitation are explored. PMID- 12562167 TI - Exercise-induced vascular remodeling. AB - Exercise produces a powerful angiogenic stimulus within the active muscle that leads to a functionally important increase in capillarity. Further, exercise can increase flow capacity by enlarging the caliber of arterial supply vessels. These adaptations are achieved by the processes of angiogenesis and arteriogenesis, respectively. PMID- 12562168 TI - Non-thermoregulatory modulation of sweating in humans. AB - Sweating in humans is critical for appropriate thermoregulation during exercise and/or exposure to warm environmental temperatures. In addition to thermal controllers of sweating, a number of non-thermal factors modulate the sweating response. This review summarizes the primary non-thermal neural modifiers of sweating in humans. PMID- 12562169 TI - Influence of estrogen on skeletal muscle damage, inflammation, and repair. AB - Estrogen may influence the disruption and diminish or delay leukocyte infiltration that occurs after muscle damage. This paper examines a mechanism by which estrogen may diminish the early postdamage migration of neutrophils into muscle and discusses the potential implications of these effects on muscle repair. PMID- 12562170 TI - Designing exercise regimens to increase bone strength. AB - Exercise is a very effective way to strengthen bones, particularly during childhood and adolescence. A collection of studies from the clinic and laboratory have provided new insights into how bone building effects of exercise can be maximized. From the available data we have calculated an "osteogenic index" for exercises. PMID- 12562171 TI - Motoneuron and sensory neuron plasticity to varying neuromuscular activity levels. AB - The size and phenotypic properties of the neural and muscular elements of the neuromuscular unit are matched under normal conditions. When subjected to chronic decreases or increases in neuromuscular activity, however, the adaptations in these properties are much more limited in the neural compared with the muscular elements. PMID- 12562172 TI - Hierarchical assembly of homochiral porous solids using coordination and hydrogen bonds. AB - A family of homochiral metal carboxylate coordination polymers have been synthesized by treating 2,2'-dihydroxy-1,1'-binaphthalene-6,6'-dicarboxylic acid (H(2)BDA) with metal salts at elevated temperatures. BDA ligands link adjacent metal centers to form 1D coordination polymeric chains using the carboxylate functionality, while the hydroxyl groups of BDA ligands form H-bonds with carboxylate oxygen atoms to link 1D coordination polymeric chains into open frameworks of higher dimensionality. We also present evidence for the important role played by H-bonds in the stabilization of open framework structures which allows for the hierarchical assembly of chiral porous solids. PMID- 12562173 TI - The fascinating noncentrosymmetric copper(II) phosphates synthesized via CsCl salt-inclusion. AB - Two new noncentrosymmetric (NCS) solids were isolated via high-temperature, salt inclusion reactions, Cs(2)Cu(7)(P(2)O(7))(4).6CsCl (1, CU-9) and Cs(2)Cu(5)(P(2)O(7))(3).3CsCl (2, CU-11). These copper(II) phosphates exhibit novel open-framework structures conceptually templated by extended Cs-Cl salt. The latter resides cooperatively in the channels upon the formation of the NCS Cu P-O frameworks, leading to the formation of fascinating salt lattices centered by the NaCl-type core. These new discoveries are significant for they may give rise to a new route for the templated synthesis of NCS solids. We are to show here the structural correlation of the newly discovered hybrid solids, and the role of the chlorine atoms in the "intergrowth" of covalent/ionic sublattices. PMID- 12562174 TI - Toward an arsenic analogue of Hittorf's phosphorus: mixed pnicogen chains in Cu(2)P(1.8)As(1.2)I(2). AB - The new compound Cu(2)P(1.8)As(1.2)I(2), with neutral, mixed pnicogen chains, has been synthesized from a stoichiometric combination of CuI, P, and As heated in vacuo at 550 degrees C. Cu(2)P(1.8)As(1.2)I(2) represents substitution of 40% of P in Cu(2)P(3)I(2) by As, with the As predominantly occupying "roof" positions in the tubular chains. The mixed pnicogen modification demonstrates increased stability (T(d) = 330 degrees C) relative to Cu(2)P(3)I(2) (T(d) = 290 degrees C) but a similar optical band gap (E(g) approximately 1.2 eV). Crystallographic data: Cu(2)P(1.8)As(1.2)I(2), monoclinic P2(1)/c, a = 15.477(3) A, b = 13.200(3) A, c = 15.421(3) A, beta = 115.406(4) degrees, Z = 16 (T = 293 K). PMID- 12562175 TI - Isolation of the novel dirhodium(II/II) thiolate compound Rh(2)(eta(1) C(6)H(5)S)(2)(mu-C(6)H(5)S)(2)(bpy)(2). AB - The reaction of the anticancer active compound [Rh(2)(mu O(2)CCH(3))(2)(bpy)(2)(CH(3)CN)(2)][BF(4)](2) (1) (bpy = 2,2'-bipyridine) with NaC(6)H(5)S under anaerobic conditions yields Rh(2)(eta(1)-C(6)H(5)S)(2)(mu C(6)H(5)S)(2)(bpy)(2).CH(3)OH (2), which was characterized by UV-visible, IR, and (1)H NMR spectroscopies as well as single-crystal X-ray crystallography. Compound 2 crystallizes as dark red platelets in the monoclinic space group C2/c with cell parameters a = 20.398(4) A, b = 11.861(2) A, c = 17.417(4) A, beta = 108.98 degrees, V = 3984.9(14) A(3), Z = 4. The main structural features are the presence of a [Rh(2)](4+) core with a Rh-Rh distance of 2.549(2) A bridged by two benzene thiolate ligands in a butterfly-type arrangement. The axial positions of the [Rh(2)](4+) core are occupied by two terminal benzene thiolates. Cyclic voltammetric studies of 2 reveal that the compound exhibits an irreversible oxidation at +0.046 V in CH(3)CN, which is in accord with the fact that the compound readily oxidizes in the presence of O(2). The fact that this unusual dirhodium(II/II) thiolate compound is formed under these conditions is an important first step in understanding the metabolism of dirhodium anticancer active compounds with thiol-containing peptides and proteins. PMID- 12562176 TI - Molecular motions in the solid state: the thermochromic nitro-nitrito Interconversion in nickel(II) bis(diamine) complexes. AB - The NO(2)(-) ion, in the trans-octahedral [Ni(II)(N,N' dimethylethylenediamine)(2)(NO(2))(2)](H(2)O) complex, coordinates the metal through the nitrogen atom (nitro form). On heating the solid complex, the anion rotates to give nitrito coordination (oxygen bound), according to a reversible process. The coordination mode of NO(2)(-) to Ni(II) is related to the steric interplay between the anion and the alkyl substituents on the diamine. PMID- 12562177 TI - Novel double insertion of carbon disulfide into two Ru-H bonds of [(dppm)(2)Ru(H)(2)] (dppm = Ph(2)PCH(2)PPh(2)): synthesis and crystal structure of a methanedithiolate complex. AB - Addition of excess carbon disulfide to cis/trans-[(dppm)(2)Ru(H)(2)] results in the methanedithiolate complex [(dppm)(2)Ru(eta(2)-S(2)CH(2))] 4 via the intermediacy of cis-[(dppm)(2)Ru(H)(SC(S)H)] 2. The X-ray crystal structure of this species has been determined. PMID- 12562178 TI - The first structurally confirmed paddlewheel compound with an M(2)(7+) core: [Os(2)(hpp)(4)Cl(2)](PF(6)). AB - Oxidation of Os(2)(hpp)(4)Cl(2), 1 (hpp = the anion of 1,3,4,6,7,8-hexahydro-2H pyrimido[1,2-a]pyrimidine), with (FeCp(2))PF(6) produces air-stable [Os(2)(hpp)(4)Cl(2)]PF(6), 2. This is the first structurally confirmed metal metal bonded paddlewheel compound having an M(2)(7+) core. The Os-Os distances for two crystalline forms, 2.2acetone and 2.hexane, are 2.3309(4) and 2.3290(6) A, respectively. EPR, (1)H NMR, and magnetization data indicate that 2 has an unpaired electron and an exceptionally low g value of 0.791 +/- 0.037. An electrochemical study shows that there is a quasireversible wave corresponding to a more highly oxidized species with an unprecedented Os(2)(8+) core. PMID- 12562179 TI - Synthesis and crystal structure of unprecedented phosphine adducts of d(1)-aryl imido-vanadium(IV) complexes. AB - The reaction of the imido precursor [V(NAr)Cl(2)](n)() (1) (Ar = 2,6-i Pr(2)C(6)H(3)) with 3 equiv of PMe(2)Ph yields the monomeric complex [V(=NAr)Cl(2)(PMe(2)Ph)(2)] (2). Reacting 1 with 1.5 equiv of dmpe or 1 equiv of dppm affords the dimeric complexes [V(=NAr)Cl(2)(dmpe)](2)(mu-P,P'-dmpe) (3) and [V(=NAr)Cl(2)(dppm)](2) (4), respectively. Complexes 2-4 have been fully characterized by spectroscopic methods, magnetism studies, and X-ray crystallography. PMID- 12562180 TI - A new metal-organic open framework consisting of threefold parallel interwoven (6,3) nets. AB - Threefold parallel interwoven (6,3) nets were assembled from Ni(II) cyclam complex and 1,3,5-tris[2-(4-carboxyphenyl)-1-ethynyl]benzene. The network generates triangular voids of effective size ca. 18.4 x 14.7 x 9.5 A. It contains 35% free volume of the crystal volume and is stable up to 300 degrees C. PMID- 12562181 TI - Electronic structure contributions to electron-transfer reactivity in iron-sulfur active sites: 1. Photoelectron spectroscopic determination of electronic relaxation. AB - Electronic relaxation, the change in molecular electronic structure as a response to oxidation, is investigated in [FeX(4)](2)(-)(,1)(-) (X = Cl, SR) model complexes. Photoelectron spectroscopy, in conjunction with density functional methods, is used to define and evaluate the core and valence electronic relaxation upon ionization of [FeX(4)](2)(-). The presence of intense yet formally forbidden charge-transfer satellite peaks in the PES data is a direct reflection of electronic relaxation. The phenomenon is evaluated as a function of charge redistribution at the metal center (Deltaq(rlx)) resulting from changes in the electronic structure. This charge redistribution is calculated from experimental core and valence PES data using a valence bond configuration interaction (VBCI) model. It is found that electronic relaxation is very large for both core (Fe 2p) and valence (Fe 3d) ionization processes and that it is greater in [Fe(SR)(4)](2)(-) than in [FeCl(4)](2)(-). Similar results are obtained from DFT calculations. The results suggest that, although the lowest energy valence ionization (from the redox-active molecular orbital) is metal based, electronic relaxation causes a dramatic redistribution of electron density ( approximately 0.7e) from the ligands to the metal center corresponding to a generalized increase in covalency over all M-L bonds. The more covalent tetrathiolate achieves a larger Deltaq(rlx) because the LMCT states responsible for relaxation are significantly lower in energy than those in the tetrachloride. The large observed electronic relaxation can make significant contributions to the thermodynamics and kinetics of electron transfer in inorganic systems. PMID- 12562182 TI - Electronic structure contributions to electron-transfer reactivity in iron-sulfur active sites: 2. Reduction potentials. AB - This study utilizes photoelectron spectroscopy (PES) combined with theoretical methods to determine the electronic structure contributions to the large reduction potential difference between [FeCl(4)](2)(-)(,1)(-) and [Fe(SR)(4)](2)( )(,1)(-) (DeltaE(0) approximately 1 V). Valence PES data confirm that this effect results from electronic structure differences because there is a similarly large shift in the onset of valence ionization between the two reduced species (DeltaI(vert) = 1.4 +/- 0.3 eV). Specific electronic contributions to DeltaI(vert) have been investigated and defined. Ligand field effects, which are often considered to be of great importance, contribute very little to DeltaI(vert) (DeltaE(LF) < -0.05 eV). By contrast, electronic relaxation, a factor that is often neglected in the analysis of chemical reactivity, strongly affects the valence ionization energies of both species. The larger electronic relaxation in the tetrathiolate allows it to more effectively stabilize the oxidized state and lowers its I(vert) relative to that of the chloride (DeltaE(rlx) = 0.2 eV). The largest contribution to the difference in redox potentials is the much lower effective charge () of the tetrathiolate in the reduced state, which results in a large difference in the energy of the Fe 3d manifold between the two redox couples (DeltaE(Fe)( )(3d) = 1.2 eV). This difference derives from the significantly higher covalency of the iron-thiolate bond, which decreases and significantly lowers its redox potential. PMID- 12562183 TI - Electronic structure contributions to electron-transfer reactivity in iron-sulfur active sites: 3. Kinetics of electron transfer. AB - The kinetics of electron transfer for rubredoxins are examined using density functional methods to determine the electronic structure characteristics that influence and allow for fast electron self-exchange in these electron-transport proteins. Potential energy surfaces for [FeX(4)](2-,1-) models confirm that the inner-sphere reorganization energy is inherently small for tetrathiolates ( approximately 0.1 eV), as evidenced by the only small changes in the equilibrium Fe-S bond distance during redox (Deltar(redox) approximately 0.05 A). It is concluded that electronic relaxation and covalency in the reduced state allow for this small in this case relative to other redox couples, such as the tetrachloride. Using a large computational model to include the protein medium surrounding the [Fe(SCys)(4)](2-,1-) active site in Desulfovibrio vulgaris Rubredoxin, the electronic coupling matrix element for electron self-exchange is defined for direct active-site contact (H0(DA)). Simple Beratan-Onuchic model is used to extend coupling over the complete surface of the protein to provide an understanding of probable electron-transfer pathways. Regions of similar coupling properties are grouped together to define a surface coupling map, which reveals that very efficient self-exchange occurs only within 4 sigma-bonds of the active site. Longer-range electron transfer cannot support the fast rates of electron self-exchange observed experimentally. Pathways directly through the two surface cysteinate ligands dominate, but surface-accessible amides hydrogen-bonded to the cysteinates also contribute significantly to the rate of electron self-exchange. PMID- 12562184 TI - Magneto-structural xorrelations in 2D and 3D extended structures of manganese(II) malonate systems. AB - Two polymeric malonato-bridged manganese(II) complexes of formula [Mn(mal)(H(2)O)(2)](n)() (1) and [Mn(2)(mal)(2)(4,4'-bipy)(H(2)O)(2)](n)() (2) have been synthesized and characterized (mal = malonate dianion; 4,4'-bipy = 4,4' bipyridine). The crystal structure of complex 1 was already known. Complex 2 crystallizes in monoclinic space group P2(1)/n, Z = 2, with unit cell parameters of a = 7.2974(10) A, b = 18.7715(10) A, c = 7.514(3) A, and beta = 91.743(12) degrees. The structure determination reveals that the complex [Mn(2)(mal)(2)(4,4' bipy)(H(2)O)(2)](n)() (2) is a 3D network being composed of Mn-malonate sheets which are pillared by bidentate 4,4'-bipy spacer forming small voids. The Mn-Mn distances through Mn-mu-(O3-C8-O4)-Mn, Mn-mu(O1-C6-O2)-Mn, and Mn-mu-4,4'-bipy-Mn bridges are 5.561, 5.410, and 11.723 A, respectively. The magnetic behaviors of complexes 1 and 2 in the temperature range 300-2 K are very close, corresponding to a weak antiferromagnetic coupling. The magnetic pathways of complex 1 are through two Mn-O-C-O-Mn with anti-anti conformation and two Mn-O-C-O-Mn with syn anti conformations and in complex 2 through all Mn-O-C-O-Mn with syn-anti conformations. Both syn-anti and anti-anti conformations create weak antiferromagnetic coupling, and the susceptibility data are fitted by the expansion series of Lines and the Curely formula for an S = 5/2 antiferromagnetic quadratic layer, based on the exchange Hamiltonian H = -Sigma(nn)()JS(i)()S(j)(). The best fit is given by the superexchange parameters J = -0.32 cm(-)(1) and g = 2.00 for complex 1 and J = -0.14 cm(-)(1), J(inter) = -0.031 cm(-)(1), and g = 2.00 for complex 2. Finally, in both the complexes there is a magnetic pathway Mn O-C-C-C-O-Mn, and this pathway through the three carbon atoms of the malonato bridging ligand could be considered negligible. PMID- 12562185 TI - Structural and spectroscopic studies of nickel(II) complexes with a library of Bis(oxime)amine-containing ligands. AB - A library of tripodal amine ligands with two oxime donor arms and a variable coordinating or noncoordinating third arm has been synthesized, including two chiral ligands based on l-phenylalanine. Their Ni(II) complexes have been synthesized and characterized by X-ray crystallography, UV-vis absorption, circular dichroism, and FTIR spectroscopy, mass spectrometry, and room temperature magnetic susceptibility. At least one crystal structure is reported for all but one Ni/ligand combination. All show a six-coordinate pseudo octahedral coordination geometry around the nickel center, with the bis(oxime)amine unit coordinating in a facial mode. Three distinct structure types are observed: (1) for tetradentate ligands, six-coordinate monomers are formed, with anions and/or solvent filling out the coordination sphere; (2) for tridentate ligands, six-coordinate monomers are formed with Ni(II)(NO(3))(2), with one monodentate and one bidentate nitrate filling the remaining coordination positions; (3) for tridentate ligands, six-coordinate, bis(mu-Cl) dimers are formed with Ni(II)Cl(2), with one terminal and two bridging chlorides filling the coordination sphere. The UV-vis absorption spectra of the complexes show that the value of 10 Dq varies according to the nature of the third arm of the ligand. The trend based on the third arm follows the order alkyl/aryl < amide < carboxylate < alcohol < pyridyl < oxime. PMID- 12562186 TI - Thallium pi-cation complexation by calix[4]tubes: (205)Tl NMR and x-ray evidence. AB - Thallium cation complexation by calix[4]tubes has been investigated by a combination of (205)Tl, (1)H NMR and ES MS demonstrating the solution formation of a dithallium complex in which the cations are held in the calix[4]arene cavities. In addition, the structure of the complex has been determined in the solid state revealing the cations to be held exclusively by pi-cation interactions. Furthermore, this crystal structure has been used as the basis for molecular dynamics simulations to confirm that binding of the smaller K(+) cation in the calix[4]tube cryptand like array occurs via the axial route featuring a pi cation intermediate. PMID- 12562187 TI - Investigation of the extraction complexes of light lanthanides(III) with bis(2,4,4-trimethylpentyl)dithiophosphinic acid by EXAFS, IR, and MS in comparison with the americium(III) complex. AB - The structure of the extraction complexes of light lanthanides (La(III), Nd(III), Eu(III)) with bis(2,4,4-trimethylpentyl)dithiophosphinic acid (HBTMPDTP) have been characterized with extended X-ray absorption fine structure spectroscopy (EXAFS), IR, and MS; the IR spectrum of the extraction complex of (241)Am with HBTMPDTP has been studied too. The molecular formula of the extraction complexes of lanthanides is deduced to be HML(4).H(2)O (M = La, Nd, Eu; L = anion of HBTMPDTP). The coordination number of Ln(III) in the complexes is 8; the coordinated donor atoms are 7 sulfur atoms from 4 HBTMDTP molecules and 1 O atom from a hydrated water molecule. With the increase of the atomic number of Ln, the coordination bond lengths of Ln-O and Ln-S decrease in the complexes. For La(III), Nd(III), and Eu(III), the coordination bond lengths of Ln-O are 2.70, 2.56, and 2.50, respectively, the coordination bond lengths of Ln-S are 3.01, 2.91, and 2.84, respectively, and the average distances between Ln and P atoms are 3.60, 3.53, and 3.46, respectively. The structure of the extraction complexes of Ln(III) with HBTMDTP is different from that of the Am(III) extraction complex. The results of IR show that there is no water coordinated with Am in the extraction complex. The molecular formula of the complex of Am(III) is deduced as being HAmL(4), and there are 8 S atoms from 4 HBTMPDTP molecules coordinated with Am. Composition and structure differences of the extraction complexes may be one of the most most important factors affecting the excellent selectivity of HBTMPDTP for Am(III) over Ln(III). PMID- 12562188 TI - Solution-state and solid-state structural characterization of complexes of a new macrocyclic ligand containing the 1,5-diazacyclooctane subunit. AB - The synthesis and characterization of a new constrained tetraazamacrocyclic ligand, 1,4,8,11-tetraazabicyclo[9.3.3]heptadecane (1,11-C(3)-cyclam), is reported. Because of its basicity, this ligand (pK(a) of the protonated form >13.5) requires aprotic solvents for its metalation reactions. Two complexes of this ligand, [Ni(1,11-C(3)-cyclam](OTf)(2) and [Co(1,11-C(3) cyclam)(NCS)(2)](OTf), have been characterized by single-crystal X-ray crystallography. For the Ni(II) complex, the 1,5-diazacyclooctane (daco) subunit of the ligand is in the chair-boat conformation, whereas that same subunit in the Co(III) complex is in the chair-chair conformation. For the Ni(II) complex, C(12) and H(12a) block one of the coordination sites. The (1)H and (13)C NMR spectra of the Ni(II) complex in D(2)O have very sharp resonances, indicative of low-spin Ni(II). The resonance for H(12a) appears at 4.5 ppm, suggesting an interaction with Ni(II). In acetonitrile, the (1)H and (13)C spectra are broadened, indicative of a low-spin/high-spin equilibrium due to axial coordination by acetonitrile. C(12) experiences the greatest degree of broadening in the (13)C NMR spectrum. Variable-temperature NMR spectroscopy from -70 to +80 degrees C shows no significant change as a function of temperature. The electronic spectrum of the Ni(II) complex (lambda(max) = 449.9 nm) is consistent with steric and electronic factors for this complex. PMID- 12562189 TI - Binuclear manganese compounds of potential biological significance. 1. Syntheses and structural, magnetic, and electrochemical properties of dimanganese(II) and (II,III) complexes of a bridging unsymmetrical phenolate ligand. AB - Reactions of the unsymmetrical phenol ligand 2-(bis(2-pyridylmethyl)aminomethyl) 6-((2-pyridylmethyl)(benzyl)aminomethyl)-4-methylphenol with Mn(OAc)(2).4H(2)O or Mn(H(2)O)(6)(ClO(4))(2) in the presence of NaOBz affords the dimanganese(II) complexes 1(CH(3)OH), [Mn(2)(L)(OAc)(2)(CH(3)OH)](ClO(4)), and 2(H(2)O), [Mn(2)(L)(OBz)(2)(H(2)O)](ClO(4)), respectively. On the other hand, reaction of the ligand with hydrated manganese(III) acetate furnishes the mixed-valent derivative 3(H(2)O), [Mn(2)(L)(OAc)(2)(H(2)O)](ClO(4))( 2). The three complexes have been characterized by X-ray crystallography. 1(CH(3)OH) crystallizes in the monoclinic system, space group P2(1)/c, with a = 10.9215(6) A, b = 20.2318(12) A, c = 19.1354(12) A, alpha = 90 degrees, beta = 97.5310(10) degrees, gamma = 90 degrees, V = 4191.7 A(3), and Z = 4. 2(H(2)O) crystallizes in the monoclinic system, space group P2(1)/n, with a = 10.9215(6) A, b = 20.2318(12) A, c = 19.1354(12) A, alpha = 90 degrees, beta = 97.5310(10) degrees, gamma = 90 degrees, V = 4191.7 A(3), and Z = 4. 3(H(2)O) crystallizes in the monoclinic system, space group P2(1)/c, with a = 11.144(6) A, b = 18.737(10) A, c = 23.949(13) A, alpha = 90 degrees, beta = 95.910(10) degrees, gamma = 90 degrees, V = 4974(5) A(3), and Z = 4. Magnetic measurements revealed that the three compounds exhibit very similar magnetic exchange interactions -J = 4.3(3) cm( )(1). They were used to establish tentative magneto-structural correlations which show that for the dimanganese(II) complexes -J decreases when the Mn-O(phenoxo) distance increases as expected from orbital overlap considerations. For the dimanganese(II,III) complexes, crystallographic results show that the Mn(II) O(phenoxo) and Mn(III)-O(phenoxo) bond lengths are inversely correlated. An interesting magneto-structural correlation is found between -J and the difference between these bond lengths, delta(Mn)(-)(O) = d(Mn)()II(-)(O) - d(Mn)()III(-)(O): the smaller this difference, the larger -J. Electrochemical studies show that the mixed-valence state is favored in 1-3 by ca. 100 mV with respect to analogous complexes of symmetrical ligands, owing to the asymmetry of the electron density as found in the analogous diiron complexes. PMID- 12562190 TI - Synthesis, crystal structure, and magnetic properties of ([Cu(oxbe)]Mn(H(2)O)[Cu(oxbe)(DMF)])(n).nDMF.nH(2)O: From dissymmetrical mononuclear entities to a 3D heterometallic supramolecular coordination polymer. AB - Self-assembly of the building block [Cu(oxbe)](-) with Mn(II) led to a novel coordination polymer ([Cu(oxbe)]Mn(H(2)O)[Cu(oxbe)(DMF)])n).nDMF.nH(2)O, where H(3)oxbe is a new dissymmetrical ligand N-benzoato-N'-(2-aminoethyl)oxamido and DMF = dimethylformamide. The crystal forms in the triclinic system, space group Ponemacr;, with a = 9.260(4) A, b = 12.833(5) A, c = 15.274(6) A, alpha = 76.18(3) degrees, beta = 82.7(3) degrees, gamma = 82.31(3) degrees, and Z = 2. The crystal structure of the title complex reveals that the two-dimensional bimetallic layers are constructed of Cu(II)Mn(II)Cu(II) chains linked together by carboxylate bridge and hydrogen bonds help to produce a novel three-dimensional channel-like structure. The magnetic susceptibility measurements (5-300 K) were analyzed by means of the Hamiltonian H = -2JS(Mn)(S(Cu1) + S(Cu2)), leading to J = -17.4 cm(-1). PMID- 12562191 TI - Structure and reactivity of a chromium(v) glutathione complex. AB - Chromium(V) glutathione complexes are among the likely reactive intermediates in Cr(VI)-induced genotoxicity and carcinogenicity. The first definitive structure of one such complex, [Cr(V)O(LH(2))(2)](3)(-) (I; LH(5) = glutathione = GSH), isolated from the reaction of Cr(VI) with excess GSH at pH 7.0 (O'Brien, P.; Pratt, J.; Swanson, F. J.; Thornton, P.; Wang, G. Inorg. Chim. Acta 1990, 169, 265-269), has been determined by a combination of electrospray mass spectrometry (ESMS), X-ray absorption spectroscopy (XAS), EPR spectroscopy, and analytical techniques. In addition, Cr(V) complexes of GSH ethyl ester (gamma-Glu-Cys GlyOEt) have been isolated and characterized by ESMS, and Cr(III) products of the Cr(VI) + GSH reaction have been isolated and characterized by ESMS and XAS. The thiolato and amido groups of the Cys residue in GSH are responsible for the Cr(V) binding in I. The Cr-ligand bond lengths, determined from multiple-scattering XAFS analysis, are as follows: 1.61 A for the oxo donor; 1.99 A for the amido donors; and 2.31 A for the thiolato donors. A significant electron withdrawal from the thiolato groups to Cr(V) in I was evident from the XANES spectra. Rapid decomposition of I in aqueous solutions (pH = 1-13) occurs predominantly by ligand oxidation with the formation of Cr(III) complexes of GSH and GSSG. Maximal half-lives of the Cr(V) species (40-50 s at [Cr] = 1.0 mM and 25 degrees C) are observed at pH 7.5-8.0. The experimental data are in conflict with a recent communication (Gaggelli, E.; Berti, F.; Gaggelli, N.; Maccotta, A.; Valensin, G. J. Am. Chem. Soc. 2001, 123, 8858-8859) on the formation of a Cr(V) dimer as a major product of the Cr(VI) + GSH reaction, which may have resulted from misinterpretation of the ESMS and NMR spectroscopic data. PMID- 12562192 TI - Ruthenium(II) complexes containing 8-(dimethylphosphino)quinoline (Me(2)Pqn): preparation, crystal structures, and electrochemical and spectroscopic properties of [Ru(bpy or phen)(3)(-)(n)(Me(2)Pqn)(n)](PF(6))(2) (bpy = 2,2'-bipyridine; phen = 1,10-phenanthroline; n = 1, 2, or 3). AB - Several new ruthenium(II) complexes containing 8-(dimethylphosphino)quinoline (Me(2)Pqn) were synthesized, and their structures and electrochemical/spectroscopic properties have been investigated. In addition to the mono(Me(2)Pqn) complex [Ru(bpy or phen)(2)(Me(2)Pqn)](PF(6))(2) (1 or 1'; bpy = 2,2'-bipyridine; phen = 1,10-phenanthroline), the geometrical isomers trans(P)- and C(1)-[Ru(bpy)(Me(2)Pqn)(2)](PF(6))(2) (tP-2 and C(1)-2) and mer- and fac [Ru(Me(2)Pqn)(3)](PF(6))(2) (m-3 and f-3) were also selectively synthesized and isolated. It was found that complexes tP-2 and m-3 were converted quantitatively to the corresponding C(1)-2 and f-3 isomers, respectively, by irradiation of light corresponding to the MLCT transition energy. The strong trans influence of the Me(2)P- donor group of Me(2)Pqn was confirmed by the X-ray structural analyses for 1, tP-2, m-3, and f-3. Cyclic voltammetry of a series of complexes, [Ru(bpy)(3)](PF(6))(2), 1, C(1)-2, and f-3, exhibited a reversible one-electron oxidation wave and two or three one-electron reduction waves. The oxidation potentials of the complexes gave a large positive shift with increasing number of coordinated Me(2)Pqn molecules, indicating a larger pi-acceptability of the Me(2)P- group compared with bpy or qn. Complex f-3 in EtOH/MeOH (4:1) glass at 77 K exhibited an intense long-lived (tau = 920 microseconds) emission arising from the quinoline-based (3)(pi-pi) excited state. In contrast, the mixed-ligand complexes 1, 1', and C(1)-2 showed a characteristic dual emission, giving a double-exponential emission decay, and the dual emission originates from both the bpy-based (3)MLCT and the quinoline-based (3)(pi-pi) emitting states. PMID- 12562193 TI - Synthesis and reactions of molybdenum triamidoamine complexes containing hexaisopropylterphenyl substituents. AB - We have synthesized a triamidoamine ligand ([(RNCH(2)CH(2))(3)N](3)(-)) in which R is 3,5-(2,4,6-i-Pr(3)C(6)H(2))(2)C(6)H(3) (hexaisopropylterphenyl or HIPT). The reaction between MoCl(4)(THF)(2) and H(3)[HIPTN(3)N] in THF followed by 3.1 equiv of LiN(SiMe(3))(2) led to formation of orange [HIPTN(3)N]MoCl. Reduction of MoCl (Mo = [HIPTN(3)N]Mo) with magnesium in THF under dinitrogen led to formation of salts that contain the ((Mo(N(2)))(-) ion. The (Mo(N(2)))(-) ion can be oxidized by zinc chloride to give Mo(N(2)) or protonated to give MoN=NH. The latter was found to decompose to yield MoH. Other relevant compounds that have been prepared include (Mo=N-NH(2))(+) (by protonation of MoN=NH), M=1;N, (Mo=NH)(+) (by protonation of M=N), and (Mo(NH(3)))(+) (by treating MoCl with ammonia). (The anion is usually (B(3,5-(CF(3))(2)C(6)H(3))(4))(-) = (BAr'(4))(-).) X-ray studies were carried out on (Mg(DME)(3))(0.5)[Mo(N(2))], MoN=NMgBr(THF)(3), Mo(N(2)), M=N, and (Mo(NH(3)))(BAr'(4)). These studies suggest that the HIPT substituent on the triamidoamine ligand creates a cavity that stabilizes a variety of complexes that might be encountered in a hypothetical Chatt-like dinitrogen reduction scheme, perhaps largely by protecting against bimolecular decomposition reactions. PMID- 12562194 TI - Peripheral metal-ion binding to tris(thia-oxo crown) porphyrazines. AB - We report the preparation and solution properties of metal-free [1b(H(2))] and cobalt [1c(Co)] porphyrazines (pz's), where three pyrroles are functionalized with a S(2)O(3) crown ether for metal-ion binding and the fourth pyrrole is appended with two long-chain alcohols (bis((11-hydroxyundecyl)thio)) for potential use as surface anchors. Compounds 1b(H(2) and 1c(Co) exhibit ion specific optical changes in the presence of Ag(+) and Hg(2+). Binding of Ag(+) to 1b(H(2)) is described by a 1:1 binding isotherm, with K(D) approximately 147 microM, whereas binding of Hg(2+) to 1b(H(2)) appears more complex. For 1c(Co), binding of Ag(+) and Hg(2+) also can be fit to a 1:1 isotherm, with K(D) approximately 109 and 83 microM, respectively. All four titrations show nonisosbesticity, including those apparently describable by the 1:1 isotherm, which indicates that there are intermediate stages as multiple ions bind. Neither 1b(H(2)) or 1c(Co) gives optical responses to Ni(2+), Zn(2+), Pb(2+), or Cu(2+) or to alkali (Li(+), Na(+), K(+), and Cs(+)) and alkaline earth (Mg(2+), Ca(2+), and Ba(2+)) metal ions. Nonetheless, "hard" ions are sensed electrochemically: Na(+) and Li(+) strongly shift the pz/pz(-) couple of 1b(H(2)()) and Co(III)/Co(II) couple of 1c(Co). For the addition of 4 equiv of Li(+) to 1c(Co), the Co(III)/Co(II) shifts +40 mV, and for the addition of 4 equiv of Na(+) to 1c(Co), the shift is +155 mV. The shifted redox waves of 1c(Co) all retain their reversibility. In contrast, for the addition of either 4 equiv of Li(+) or Na(+) to 1b(H(2)), the shifts of the pz/pz(-) couple are essentially the same, approximately 170 mV, and the shifted redox waves become broadened and less reversible, due to ion-induced aggregation. For 1c(Co) the shape of the titration curves [M]/[1b(H(2))] versus E(1/2) is concave to the x-axis, implying cooperative binding of multiple M(+) ions as part of the redox/binding system of equilibria. PMID- 12562195 TI - New heterometallic coordination polymers self-assembled from copper(II) nitrate and (diamine)Pt(II)(pyridinecarboxylates)(2). AB - One-dimensional (1-D), two-dimensional (2-D), and three-dimensional (3-D) coordination polymers were prepared by self-assembly of binary metal complex systems, copper(II) nitrate and (en)Pt(II)(nic)(2) or (dmpda)Pt(II)(isonic)(2) (en = ethylenediamine, dmpda = 2,2'-dimethyl-1,3-propanediamine, nic = nicotinate, and isonic = isonicotinate), in aqueous solutions. Equimolar reactions of copper(II) nitrate with (dmpda)Pt(II)(isonic)(2) and (en)Pt(II)(nic)(2) resulted in 1-D ([(dmpda)Pt(isonic)(2)Cu(OH(2))(3)](NO(3))(2))(n)() (1) and 2-D ([(en)Pt(nic)(2)Cu(OH(2))](NO(3))(2))(n) (2), respectively, but the reaction of (en)Pt(II)(nic)(2) with excess copper(II) nitrate gave 3-D ([((en)Pt(nic)(2))(3)Cu(5)(OH)(2)(OH(2))(6)](NO(3))(8))(n) (3). The local structure of crystal 1 has a mononuclear copper unit, 2 has a dinuclear copper unit with a Cu-Cu distance of 2.659(5) A, and 3 has a pentanuclear copper unit. The methyl groups of the dmpda ligand are located in the space between two isonicotinate ligands of 1, which is presumed to be an important factor to determine the final structure of the crystal formed by self-assembly. Magnetic behaviors of crystals 1-3 examined in the temperature range of 4-300 K appear to be governed by the local structures around the copper(II) ions and do not indicate any significant long-range magnetic exchange interactions along the polymeric chain. PMID- 12562196 TI - The effect of a 3-benzyl group on the coordination chemistry of homoscorpionate ligands. AB - New homoscorpionate ligands containing a 3-benzyl substituent, hydrotris(3-benzyl 5-methylpyrazol-1-yl)borate, Tp(Bn,Me), and hydrotris(3-benzyl-4-phenylpyrazol-1 yl)borate, Tp(Bn,4Ph), have been synthesized, and the dynamic behavior of a number of metal complexes was studied by NMR. Structures of the complexes Tl[Tp(Bn,Me)], 1, Tl[Tp(Bn,4Ph)], 2, Co[Tp(Bn,Me)][Tp(Np)], 3, Mo[Tp(Bn,Me)](CO)(2)NO, 4, Co[Tp(Bn,4Ph)][Tp], 5, and Mo[Tp(Bn,Me)](CO)(2)(eta(3) methallyl), 6, were determined by X-ray crystallography. In the Tp(Bn,Me) ligand, the benzyl group is freely rotating and provides less steric hindrance to the coordinated metal than a neopentyl group, but steric hindrance is increased in the Tp(Bn,4Ph) ligand, where the rotation of the benzyl substituent is restricted by the 4-phenyl substituent. PMID- 12562197 TI - Factors affecting the electrochemical and spectroelectrochemical properties of diruthenium(III,II) complexes containing four identical unsymmetrical bridging ligands. AB - Factors affecting the electrochemical and spectroelectrochemical properties of diruthenium(III,II) complexes containing four unsymmetrical bridging ligands are reported for seven related compounds which were isolated in one or two of the four possible isomeric forms. The investigated compounds are represented as Ru(2)(2-CH(3)ap)(4)Cl, Ru(2)(2,5-F(2)ap)(4)Cl, Ru(2)(2,6-F(2)ap)(4)Cl, and Ru(2)(2,4,6-F(3)ap)(4)Cl where 2-CH(3)ap, 2,5-F(2)ap, 2,6-F(2)ap, and 2,4,6 F(3)ap are, respectively, the 2-(2-methylanilino)pyridinate anion, the 2-(2,5 difluoroanilino)pyridinate anion, the 2-(2,6-difluoroanilino)pyridinate anion, and the 2-(2,4,6-trifluoroanilino)pyridinate anion. Ru(2)(2-CH(3)ap)(4)Cl and Ru(2)(2,5-F(2)ap)(4)Cl exist only in a (4,0) conformation while Ru(2)(2,4,6 F(3)ap)(4)Cl is present in both (3,1) and (4,0) isomeric forms. Ru(2)(2,6 F(2)ap)(4)Cl also exists in two isomeric forms, but only the (3,1) isomer was generated in sufficient quantities to be isolated and structurally characterized. This series of seven closely related metal-metal bonded complexes thus provides the first possibility to systematically examine how differences in position and number of the electron-donating or electron-withdrawing groups on the anionic bridging ligands might be related to the electronic properties and structural features of the compound as well as the type and number of geometric isomers which are formed. Each diruthenium derivative undergoes three one-electron transfers in CH(2)Cl(2) containing 0.1 M tetra-n-butylammonium perchlorate (TBAP). The first reduction and first oxidation products were characterized by thin-layer UV-vis spectroelectrochemistry, and the spectroscopic data, along with E(1/2) values, were then related via linear free energy relationships to the type of isomer and/or position of the electron-donating or electron-withdrawing substituents on the anionic ap bridge. The electrogenerated Ru(2)(6+) and Ru(2)(4+) forms of the compounds were assigned on the basis of electrochemical and UV-vis spectroscopic data as having the electronic configuration sigma(2)pi(4)delta(2)pi(2) and sigma(2)pi(4)delta(2)pi(3)delta, respectively, and seemed to be independent of the isomer type ((3,1) or (4,0)). The spectral and electrochemical properties of the compounds both vary substantially as a function of the isomer type, but this is not reflected in the structural features of the compounds which are within the range of what is seen for other Ru(2)(5+) species described in the literature. The Ru-Ru bond lengths of the four structurally characterized (4,0) isomers of the ap complexes range from 2.275 to 2.296 A while those of the three structurally characterized (3,1) isomers of ap derivatives fall in the range 2.284-2.286 A and show no significant difference among the three compounds. The Ru-Cl bond lengths of the (3,1) isomers do not vary significantly with the bridging ligand and range from 2.458 to 2.471 A whereas those of the (4,0) isomers range from 2.437 to 2.487 A and show larger variations among the compounds. The Ru-Ru-Cl bond angle is virtually independent of the bridging ligand in the case of the (4,0) isomers but decreases with the electron withdrawing effect of the substituent in the case of the (3,1) isomers. PMID- 12562198 TI - Subtle role of polyatomic anions in molecular construction: structures and properties of AgX bearing 2,4'-thiobis(pyridine) (X(-) = NO(3)(-), BF(4)(-), ClO(4)(-), PF(6)(-), CF(3)CO(2)(-), and CF(3)SO(3)(-)). AB - Studies on the subtle effects and roles of polyatomic anions in the self-assembly of a series of AgX complexes with 2,4'-Py(2)S (X(-) = NO(3)(-), BF(4)(-), ClO(4)( ), PF(6)(-), CF(3)CO(2)(-), and CF(3)SO(3)(-); 2,4'-Py(2)S = 2,4' thiobis(pyridine)) have been carried out. The formation of products appears to be primarily associated with a suitable combination of the skewed conformers of 2,4' Py(2)S and a variety of coordination geometries of Ag(I) ions. The molecular construction via self-assembly is delicately dependent upon the nature of the anions. Coordinating anions afford the 1:1 adducts [Ag(2,4'-Py(2)S)X] (X(-) = NO(3)(-) and CF(3)CO(2)(-)), whereas noncoordinating anions form the 3:4 adducts [Ag(3)(2,4'-Py(2)S)(4)]X(3) (X(-) = ClO(4)(-) and PF(6)(-)). Each structure seems to be constructed by competition between pi-pi interactions of 2,4'-Py(2)S spacers vs Ag.X interactions. For ClO(4)(-) and PF(6)(-), an anion-free network consisting of linear Ag(I) and trigonal Ag(I) in a 1:2 ratio has been obtained whereas, for the coordinating anions NO(3)(-) and CF(3)CO(2)(-), an anion-bridged helix sheet and an anion-bridged cyclic dimer chain, respectively, have been assembled. For a moderately coordinating anion, CF(3)SO(3)(-), the 3:4 adduct [Ag(3)(2,4'-Py(2)S)(4)](CF(3)SO(3))(3) has been obtained similarly to the noncoordinating anions, but its structure is a double strand via both face-to face (pi-pi) stackings and Ag.Ag interactions, in contrast to the noncoordinating anions. The anion exchanges of [Ag(3)(2,4'-Py(2)S)(4)]X(3) (X(-) = BF(4)(-), ClO(4)(-), and PF(6)(-)) with BF(4)(-), ClO(4)(-), and PF(6)(-) in aqueous media indicate that a [BF(4)(-)] analogue is isostructural with [Ag(3)(2,4' Py(2)S)(4)]X(3) (X(-) = ClO(4)(-) and PF(6)(-)). Furthermore, the anion exchangeability for the noncoordinating anion compounds and the X-ray data for the coordinating anion compounds establish the coordinating order to be NO(3)(-) > CF(3)CO(2)(-) > CF(3)SO(3)(-) > PF(6)(-) > ClO(4)(-) > BF(4)(-). PMID- 12562199 TI - Theoretical study of mixed MLaX(4) (M = Na, K, Cs; X = F, Cl, Br, I) rare earth/alkali metal halide complexes. AB - The structure, bonding, and vibrational properties of the mixed MLaX(4) (M = Na, K, Cs; X = F, Cl, Br, I) rare earth/alkali metal halide complexes have been studied using the MP2 method in conjunction with polarized triple-zeta valence basis sets and quasi-relativistic effective core potentials for the heavy atoms. From the three characteristic structures, possessing 1- (C(3)(v)), 2- (C(2)(v)), or 3-fold coordination (C(3)(v)) between the alkali metal and the bridging halide atoms, the bi- and tridentate forms are stable isomers with close dissociation energies. In general, for the complexes existing of lighter alkali metals and halogens, the bidentate structure corresponds to the global minimum of the potential energy surface, while the heavier analogues favor the tridentate structure. At experimentally relevant temperatures (T > 800 K), however, the isomerization entropy leads to a domination of the bidentate structures over the tridentate forms for all complexes. An important effect of the size of the alkali metal is manifested in the larger stabilities of the K and Cs complexes. The natural atomic charges are in agreement with strong electrostatic interactions in the title complexes. The marginal covalent contributions show a slight increasing trend in the heavier analogues. The calculated vibrational data indicate that infrared spectroscopy may be an effective tool for experimental investigation and characterization of MLaX(4) molecules. PMID- 12562200 TI - Spectroscopic and computational studies of a Ni(+)-CO model complex: implications for the acetyl-CoA synthase catalytic mechanism. AB - The four-coordinate Ni(+) complex [PhTt(t)(Bu)]Ni(I)CO, where PhTt(t)()(Bu) = phenyltris((tert-buthylthio)methyl)borate (a tridentate thioether donor ligand), serves as a possible model for key Ni-CO reaction intermediates in the acetyl-CoA synthase (ACS) catalytic cycle. Resonance Raman, electronic absorption, magnetic circular dichroism (MCD), variable-temperature variable-field MCD, and electron paramagnetic resonance spectroscopies were utilized in conjunction with density functional theory and semiemperical INDO/S-CI calculations to investigate the ground and excited states of [PhTt(t)()(Bu)]Ni(I)CO. These studies reveal extensive Ni(+) --> CO pi-back-bonding interactions, as evidenced by a low C-O stretching frequency (1995 cm(-)(1)), a calculated C-O stretching force constant of 15.5 mdyn/A (as compared to k(CO)(free CO) = 18.7 mdyn/A), and strong Ni(+) - > CO charge-transfer absorption intensities. Calculations reveal that this high degree of pi-back-bonding is due to the fact that the Ni(+) 3d orbitals are in close energetic proximity to the CO pi acceptor orbitals. In the ACS "paramagnetic catalytic cycle", the high degree of pi-back-bonding in the putative Ni(+)-CO intermediate (the NiFeC species) is not expected to preclude methyl transfer from CH(3)-CoFeSP. PMID- 12562201 TI - Century-known copper salt Cu(OAc)(OMe) proven to be a unique magnetic lattice composed of tetranuclear copper(II) species with a rare binding mode of the acetate anion. AB - The structure, spectroscopy, and magnetism of a century-old copper salt, Cu(OAc)(OMe), is reported. The crystal structure contains two independent Cu(II) ions, which are both five-coordinated and which are bridged by methoxo and acetate anions to form an infinite 2D network. Thereby the methoxo groups connect Cu1 and Cu2 with their symmetry-generated counterparts Cu1(i) and Cu2(i), respectively, resulting in Cu.Cu distances of 2.9803(10) and 2.9874(10) A. Cu1 and Cu2 themselves are bridged via the carboxylate groups of two acetates leading to a Cu1.Cu2 distance of 2.9473(7) A. The tetranuclear units thus generated are cross-linked via acetate oxygens to form a 2D sheet structure. One of the two independent acetate ligands has a rare binding mode, whereby it acts as a tetradentate syn-anti, syn-anti bridging ligand. The temperature dependence of the magnetic susceptibility was assigned to be dominated by the very strong antiferromagnetic exchange coupling via bis(mu-methoxo) bridges (J(1) = -409(1) cm(-)(1)). PMID- 12562202 TI - Synthesis and characterization of chiral tetraaza macrocyclic nickel(II) and palladium(II) complexes. AB - Chiral tetraaza macrocyclic nickel(II) and palladium(II) complexes 2a-e, containing one or two (R,R)-(-)-1,2-cyclohexanediyl bridges, were synthesized by template condensation reactions and characterized by (1)H and (13)C NMR, IR, UV vis, and mass spectrometry. The electrophilic reactivity of 2a was explored. Crystal structures of Ni complex 2b and metal-free ligand 5 were determined by single-crystal X-ray diffraction. PMID- 12562203 TI - Stabilization of duplex DNA structure and suppression of transcription in vitro by bis(quinone diimine) complexes of rhodium(III) and ruthenium(II). AB - The ability of octahedral complexes possessing quinone diimine ligands to inhibit transcription by stabilization of the DNA duplex structure was investigated. Rh(III) and Ru(II) complexes possessing two quinone diimine ligands in their coordination sphere were found to significantly increase the melting temperature (DeltaT(m)) of a 15-mer duplex DNA. [Rh(phi)(2)phen](3+) and [Ru(phi)(2)phen](2+) (phi = 9,10-phenanthrenequinone diimine, phen = 1,10-phenanthroline) exhibit DeltaT(m) values of +21 and +15 degrees C relative to free 15-mer duplex (T(m) = 55 degrees C) at [complex]/[DNA bases] = 0.067 (two complexes/duplex). Similarly, a shift in the melting temperature of +14 degrees C was measured for [Rh(bqdi)(2)phen](3+) (bqdi = 1,2-benzoquinone diimine), which possesses the nonintercalating bqdi ligand. In contrast, [Ru(phen)(2)phi](2+) and [Rh(phen)(2)L](3+) (L = phi, bqdi), which possess a single quinone diimine ligand, the parent [Ru(phen)(3)](2+) and [Rh(phen)(3)](3+) complexes, and ethidium bromide result in small shifts in the melting temperature of the duplex oligonucleotide. A distinct correlation between DeltaT(m) and the relative concentration of each complex required to inhibit 50% of the transcription (R(inh)(50)) was observed, independent of the presence of an intercalative ligand. The duplex stabilization by bis(quinone diimine) complexes results in inhibition of transcription in vitro at significantly lower complex concentrations than for the corresponding [Ru(phen)(2)phi](2+) and [Rh(phen)(2)L](3+) (L = phi, bqdi) complexes. Possible explanations for these observations are discussed. PMID- 12562204 TI - Five different fluxional processes in polyfluorophenyl palladium(II) complexes with 2,4,6-tris(3,5-dimethylpyrazol-1-yl)-1,3,5-triazine. The driving effect of the solvent. AB - The polydentate N-donor ligand 2,4,6-tris(3,5-dimethylpyrazol-1-yl)-1,3,5 triazine (Me(2)-TpzT) has been used to synthesize the new palladium derivatives Pd(R)(2)(Me(2)-TpzT), R = C(6)F(5), 1; R = m-C(6)ClF(4), 2. In the case of complex 2, four different atropisomers have been detected at low temperature. The new complexes exhibit a rich dynamic behavior, including three metallotropic processes (metal-hurdling, 1,4-metallotropic shifts, and an intermolecular process) and two processes involving restricted rotation of aromatic rings (the polyfluorophenyl groups and the uncoordinated pyrazole group adjacent to the metal fragment). The fluxional behavior has been studied by (1)H and (19)F NMR spectroscopy using variable temperature NMR studies and (1)H,(1)H and (19)F,(19)F EXSY experiments. The study of solutions of 2 in 1,1',2,2'-tetrachloroethane-d(2) gave the following order for the free energy of activation: pyrazole rotation < 1,4-metallotropic shift < intermolecular exchange < polyfluorophenyl rotation. The process of metal hurdling was not found in this solvent. However, in acetone d(6) such a process was detected and was found to be of lower energy than the 1,4 metallotropic shift. In dilute acetone-d(6) or 1,1',2,2'-tetrachloroethane-d(2 )()solutions, the intermolecular process was not observed. Conclusions concerning the different mechanisms have been deduced from the data obtained. PMID- 12562205 TI - A Route to 1,2,4-oxadiazoles and their complexes via platinum-mediated 1,3 dipolar cycloaddition of nitrile oxides to organonitriles. AB - A significant activation of the Ctbd1;N group in organonitriles upon their coordination to a platinum(IV) center has been found in the reaction of [PtCl(4)(RCN)(2)] (R = Me, Et, CH(2)Ph) with the nitrile oxides 2,4,6 R'(3)C(6)H(2)CNO (R' = Me, OMe) to give the (1,2,4-oxadiazole)platinum(IV) complexes (R = Me, R' = Me (1); R = Et, R' = Me (2); R = Et, R' = OMe (3); R = CH(2)Ph, R' = Me (4)); the [2 + 3] cycloaddition was performed under mild conditions (unless poor solubility of [PtCl(4)(RCN)(2)] precludes the reaction) starting even from complexed acetonitrile and propionitrile, which exhibit low reactivity in the free state. The reaction between complexes 2-4 and 1 equiv of Ph(3)P=CHCO(2)Me in CH(2)Cl(2) leads to the appropriate platinum(II) complexes (5 7); the reduction failed only in the case of 1 insofar as this complex is insoluble in the most common organic solvents. All the platinum compounds were characterized by elemental analyses, FAB mass spectrometry, and IR and (1)H, (13)C((1)H), and (195)Pt NMR spectroscopies, and three of them also by X-ray crystallography. The oxadiazoles formed in the course of the metal-mediated reaction were liberated almost quantitatively from their Pt(IV) complexes by reaction of the latter (complexes 2-4) with an excess of pyridine in chloroform, giving free 1,2,4-oxadiazoles and trans-[PtCl(4)(pyridine)(2)]; the sequence of the Pt(IV)-mediated [2 + 3] cycloaddition and the liberation opens up an alternative route for the preparation of this important class of heterocycles. PMID- 12562206 TI - Synthesis and characterization of transition-metal zintl phases: K(10)NbInAs(6) and K(9)Nb(2)As(6). AB - The two title compounds were prepared by direct reactions of the corresponding elements at high temperature. The structures were determined by single-crystal X ray diffraction: K(10)NbInAs(6), monoclinic, P2(1)/n, Z = 2, a = 9.107(1) A, b = 8.2878(8) A, c = 15.139(1) A, beta = 91.112(9) degrees; K(9)Nb(2)As(6), monoclinic, P2(1)/c, Z = 2, a = 9.348(1) A, b = 9.113(1) A, c = 12.798(1) A, beta = 95.98(1) degrees. They contain isolated dimers made of edge-sharing tetrahedra of [NbAs(4)] and [InAs(4)] in the former, NbInAs(6)(10)(-), and only [NbAs(4)] in the latter, Nb(2)As(6)(9)(-). Magnetic measurements show that K(10)NbInAs(6) is diamagnetic, i.e., a d(0) transition-metal Zintl phase, while K(9)Nb(2)As(6) exhibits a Curie-Weiss behavior consistent with the presence of one unpaired electron. The latter defines K(9)Nb(2)As(6) as a mixed-valence (presumably of type III) transition-metal Zintl phase, only the third example of such phases. PMID- 12562207 TI - Hydrothermal syntheses and crystal structures of complex-linked three-Dimensional coordination vanadium selenites: M(4,4'-bipy)(H(2)O)V(2)Se(2)O(10) (M = Co, Ni). AB - Two inorganic-organic hybrid compounds with the formula M(4,4' bipy)(H(2)O)V(2)Se(2)O(10) (M = Co, Ni) were hydrothermally synthesized and characterized by single-crystal X-ray diffraction. Compounds Co(4,4' bipy)(H(2)O)V(2)Se(2)O(10) (1) and Ni(4,4'-bipy)(H(2)O)V(2)Se(2)O(10) (2), which are structural analogues, crystallize in the triclinic space group Ponemacr; with crystal data a = 7.9665(3) A, b = 8.1974(3) A, c = 13.8096(4) A, alpha = 85.704(2) degrees, beta = 73.5180(10) degrees, gamma = 75.645(2) degrees, V = 837.76(5) A(3), and Z = 2 and a = 7.9489(19) A, b = 8.128(2) A, c = 13.709 A, alpha = 85.838(6) degrees, beta = 73.736(8) degrees, gamma = 75.594(9) degrees, V = 823.5(4) A(3), and Z = 2, respectively. [M(4,4'-bipy)(H(2)O)V(2)Se(2)O(10)] (M = Co, Ni) have a three-dimensional structure and consist of two subunits, [(VO(2))(SeO(3))](-) infinite chains and [M(4,4'-bipy)(H(2)O)](2+) fragments. The [(VO(2))(SeO(3))](-) chains are composed of [V(2)Se(4)O(14)](4)(-) clusters linked by VO(4)N triangular bipyramids. The 4,4'-bipy molecule as a bifunctional organic ligand is directly linked to Co or Ni and V atoms, affording the three dimensionality. The compounds were characterized by infrared spectroscopy and differential thermal and thermogravimetric analyses. PMID- 12562208 TI - Synthesis, crystal structures, and third-order nonlinear optical properties of a series of ferrocenyl organometallics. AB - Three novel ferrocenyl complexes [Zn(4-PFA)(2)(NO(3))(2)](H(2)O) (1), [Hg(2)(OAc)(4)(4-BPFA)(2)](CH(3)OH) (2), and [Cd(2)(OAc)(4)(4-BPFA)(2)] (3) (4 PFA = [(4-pyridylamino)carbonyl]ferrocene, 4-BPFA = 1,1'-bis[(4 pyridylamino)carbonyl]ferrocene) were prepared, and complexes 1 and 2 were structurally characterized by means of X-ray single-crystal diffraction. In complex 1, the zinc(II) atom is coordinated at a distorted tetrahedral environment by two nitrogen atoms from two 4-PFA moieties and two oxygen atoms from two nitrate anions; [Zn(4-PFA)(2)(NO(3))(2)] units are linked by hydrogen bonds N-H.O and O-H.O forming one-dimensional chains. Complex 2 is a tetranuclear macrocycle compound consisting of two 4-BPFA moieties and two Hg atoms; [Hg(2)(OAc)(4)(4-BPFA)(2)] units form 1-D chains by hydrogen bonds N-H.O as complex 1. Some complexes with 1,1'-bisubstituted pyridine-containing ferrocene ligands have been described, but their crystal data are limited. Compound 2 is the first example of a macrocyclic pyridine-containing ferrocenyl complex. The third-order nonlinear optical (NLO) properties of 4-PFA, 4-BPFA, and complexes 1 3 were determined by Z-scan techniques. The results indicate that all the compounds exhibit strong self-focusing effect. The hyperpolarizability gamma values are calculated to be in the range 1.51 x 10(-)(28) to 3.12 x 10(-)(28) esu. The gamma values are nearly twice as large for complexes 1-3 as for their individual ligands, showing that the optical nonlinearity of the complexes is dominated by the ligands. PMID- 12562209 TI - Large-scale synthesis of tungsten oxide nanowires with high aspect ratio. AB - Through the controlled removal of surfactant from the presynthesized mesolamellar precursor (WO-L) at elevated temperature, tungsten oxide nanowires with diameters ranging from 10 to 50 nm and lengths up to several micrometers were obtained on a large scale. The structure, morphology, and composition of the nanowires were characterized by the XRD, TEM, HRTEM, EDX, and Raman spectra. PMID- 12562210 TI - Modeling proteome networks with range-dependent graphs. AB - In this paper we consider the problem of characterizing and modeling large-scale protein-protein association networks using a class of range-dependent graphs which possess appropriate small world properties. These graphs may be employed in representing given association network using a maximum likelihood approach. This in turn annotates every observed association with its 'range', representing the tendency for such an association to be transitive. The application of a very rapidly developing field of graph theory to the emerging field of proetemics is novel and allows for a many-to-many relationship between individual proteins and groupings of proteins, which in turn may correspond to distinct functional behavior. PMID- 12562211 TI - Genetic polymorphisms of estrogen receptor-alpha: possible implications for targeted osteoporosis therapy. AB - Genetic factors play an important role in the determination of bone mass and osteoporosis. A number of candidate genes have been implicated in osteoporosis, including genes encoding type 1 collagen, vitamin D receptor, estrogen receptor alpha (ERalpha), and others. A number of association studies have been performed with single nucleotide polymorphisms in the ERalpha gene to assess their relation with bone mineral density in pre- and postmenopausal women, as well as the rate of bone loss after menopause and skeletal response to estrogen administration. The polymorphisms studied thus far mostly involved intronic polymorphisms in intron 1. Other less frequently studied polymorphisms include those in exons 1, 4, and 8. Although most studies demonstrated associations with various bone related parameters, the results are still disputed. Assessing genetic factors including ERalpha polymorphisms, if their significances are confirmed, can be helpful in targeting preventive measures to individuals with higher risk of developing osteoporosis and render the preventive effort more cost-effective. Moreover, pharmacogenetically, it may help identify postmenopausal women who tend to have better skeletal responses after estrogen replacement. It is not known, however, if patients who possess favorable polymorphisms in terms of skeletal responsiveness will also have an undesirably higher risk of adverse effects. This issue needs to be further investigated before clinical decisions based on the balance between benefits and risks can be made. PMID- 12562212 TI - Catechol-O-methyl transferase (COMT) inhibitors in patients with Parkinson's disease: is COMT genotype a useful indicator of clinical efficacy? AB - In clinical practice, two potent and selective catechol-O-methyl transferase (COMT) inhibitors are available for the control of motor fluctuation in patients with Parkinson's disease. However, because of the complexity of fluctuating motor symptoms, it is difficult to evaluate the clinical efficacy of COMT inhibitors in each individual. Therefore, an objective factor predicting the clinical efficacy of COMT inhibitors is needed. Individual variation in COMT activity is regulated by a single nucleotide of the COMT gene on the long arm of chromosome 22. Therefore, there could be a correlation between COMT genotype and the clinical efficacy of COMT inhibitors. Three double-blind studies evaluating the efficacy of a single or repeated doses of a COMT inhibitor failed to find significant difference in the improvement in the duration of daily 'on' time and degree of motor abilities between patients with different COMT genotypes. Furthermore, there were no significant differences in the severity and frequency of dopaminergic adverse effects between patients with different COMT genotypes. These data suggest that the COMT genotype is not a major factor in deciding the clinical efficacy of COMT inhibitors. PMID- 12562213 TI - Predicting response to lithium in mood disorders: role of genetic polymorphisms. AB - Lithium is considered to be the first choice mood stabilizer in recurrent mood disorders. Its widespread and large-scale use is the result of its proven efficacy. In spite of this fact, patients have been observed to show a variable response to lithium treatment: in some cases it is completely effective in preventing manic or depressive relapses, while in other cases it appears to show no influence on the disease course. The possible definition of a genetic liability profile for adverse effects and efficacy will be of great help, as lithium therapy needs at least 6 months to be effective in stabilizing mood disorders. During the last few years, a number of groups have reported possible liability genes. Lithium long-term prophylactic efficacy has been associated with serotonin transporter protein, tryptophan hydroxylase and inositol polyphosphate 1-phosphatase variants. A number of other candidate genes and anonymous markers did not yield positive associations. Therefore, even if some positive results have been reported, no unequivocal susceptibility gene for lithium efficacy has been identified. Although the available data may not currently allow a meaningful prediction of lithium response, future research is aimed at the development of individualized treament of mood disorders, including the possibility of 'pharmacological genetic counseling'. PMID- 12562214 TI - Targeting mature T cell leukemia: new understanding of molecular pathways. AB - The best studied T cell leukemia/lymphoma from a genetic and biochemical point of view is T-cell chronic lymphocytic/prolymphocytic leukemia (T-CLL/T-PLL). This neoplasia commonly shows chromosomal rearrangements at 14q32.1 including translocations [t(14;14)(q11;q32), t(7;14)(q35;q32)], and inversions [inv(14)(q11;q32)]. The investigation of the locus in question at 14q32.1 resulted in the identification of two related genes named T cell leukemia/lymphoma 1 (TCL1) and TCL1b. Both genes are activated in T-CLL/T-PLL by the chromosomal aberrations mentioned above. Mice from a transgenic mouse strain expressing the TCL1 gene under the thymocyte specific lck promoter developed a mature T cell leukemia late in life, thereby demonstrating that over-expression of TCL1 induces the neoplastic transformation of T cells. Biochemically, Tcl1 protein works as a co-factor of the Akt kinase, a key regulator of antiapoptotic and proliferative signals. Tcl1 interacts physically with Akt, increases its kinase activity and facilitates its transport to the nucleus. The pathogenesis of T-CLL/T-PLL may also involve Nur77, a T cell transcription factor required for T cell receptor-mediated apoptosis. Akt phosphorylates Nur77, thereby blocking its DNA-binding ability and rendering the transcription factor inactive. The recently emerged insights into the molecular mechanisms of T cell leukemogenesis will allow for the development of specific pharmacological tools for the treatment of these hematopoietic malignancies. PMID- 12562215 TI - Genetic variation of human UDP-glucuronosyltransferase: implications in disease and drug glucuronidation. AB - The uridine diphosphate (UDP)-glucuronosyltransferases (UGTs) are key enzymes in human detoxication of xeno- and endobiotics. Potentially toxic endogenous compounds such as bilirubin, or exogenous compounds such as drugs, pesticides, and carcinogens, are generally transformed into water-soluble glucuronides for excretion in bile and urine. The UGTs are encoded by a multigene family in humans. A relatively small number of human enzymes catalyze the glucuronidation of thousands of compounds. Genetic variations and single nucleotide polymorphisms (SNPs) within the UGT genes are remarkably common, and lead to genetic polymorphisms. The multiplicity of transferases, some exhibiting overlapping substrate specificity, may provide functional compensation for genetic deficit in some cases. Genetic variation may cause different phenotypes by affecting expression levels or activities of individual UGTs. This inter-individual variation in UGTs has resulted in functional deficit affecting endogenous metabolism and leading to jaundice and other diseases. Disruption of the normal metabolic physiology, by the reduction of bile acid excretion or steroid glucuronidation, may lead to cholestasis and organ dysfunction. Deficient glucuronidation of drugs and xenobiotics have an important pharmacological impact, which may lead to drug-induced adverse reactions, and even cancer. Additional novel polymorphisms in this gene family are yet to be revealed and studied, but will have a profound effect on the development of new drugs and therapies. PMID- 12562216 TI - Baculovirus vectors: novel mammalian cell gene-delivery vehicles and their applications. AB - Various methods have been developed to transfer and express genes in mammalian cells. Each method, whether virally, non-virally, or physically-based, has unique favorable features, but also drawbacks with respect to meeting desired and specific needs. Baculoviruses have been used since 1983 to express recombinant genes controlled by strong insect-virus promoters in their natural host (insect) cells. Today this is a well-established and easy to handle system for producing large quantities of recombinant proteins for numerous purposes. In 1995 it was first published that recombinant baculoviruses are able to deliver genes into mammalian cells. These genes are expressed provided that they are controlled by a promoter which is active in mammalian cells. Since then, various vector variants have been developed and numerous potential and meaningful applications have been described. It is not surprising that the use of baculovirus vectors as mammalian cell gene delivery vectors is constantly increasing and that the system is undergoing permanent improvements. Based on the convenience of the system to transfer genes into mammalian cells, baculoviruses can be applied in cell-based assays for drug screening to overcome the long periods of time required to generate stable cell lines. Baculovirus vectors are able to deliver very large DNA sequences into mammalian cells and vectors for toxic gene products can also be generated. In addition, baculoviruses are valuable tools for launching viral infection in cases where there is no appropriate cell culture system. Moreover, recent research has shown that the vectors can be applied in vivo. Depending on the design of the study, baculovirus vectors allow for sustained gene expression or are able to induce an immune response directed against the delivered and/or displayed gene product. The latter offers the opportunity to generate monoclonal antibodies against certain proteins that have failed by other means. In addition, it points to the potential usefulness of baculovirus vectors as new kinds of vaccines. Baculovirus vectors are therefore considered an enabling technology for various product opportunities. PMID- 12562217 TI - Gene and protein domain-specific patterns of genetic variability within the G protein coupled receptor superfamily. AB - INTRODUCTION: Guanine nucleotide binding proteins (G-proteins) represent the targets for >50% of all therapeutics. There is substantial interindividual variation in response to agonists and antagonists directed to these receptors, which may, in part, be due to genetic polymorphisms. As a class, the sequence variability of G-protein-coupled receptor (GPCR) genes has not been characterized. STUDY DESIGN: This variability was investigated by sequencing promoter, 5'- and 3'-UTR, coding blocks, and intron-exon boundaries, of 64 GPCR genes in an ethnically diverse group of 82 individuals. RESULTS: Of the 675 single-nucleotide variations found, 61% occurred in > or =1% of the population sample and the nature of these 412 single nucleotide polymorphisms (SNPs) was assessed. 5'-UTR (p = 0.002) and coding (p = 0.006) SNPs were observed more often in GPCR genes, compared with 309 non-GPCR genes similarly interrogated. The prevalence of non-synonymous coding SNPs was unexpectedly high, with 65% of GPCR genes having at least one. Intron-containing genes had half as many non synonymous coding SNPs compared with intronless genes (p = 0.0009), suggesting that when introns are not available coding regions provide sites for variation. A distinct relationship between the prevalence of non-synonymous SNPs and receptor structural domains was evident (p = 0.0006 by ANOVA), with variability being most prominent in the transmembrane spanning domains (38%) and the intracellular loops (24%). Phosphoregulatory domains, particularly the carboxy terminus, often the site for agonist-promoted phosphorylation by G-protein coupled receptor kinases, were the least polymorphic (8%). CONCLUSIONS: There is substantial genetic variability in potentially pharmacologically relevant coding and noncoding regions of GPCRs. Such variability should be considered in the development of new agents, or optimization of existing agents, targeted to these receptors. PMID- 12562218 TI - Alphabet soup: what's the meaning of all those degrees and credentials?? PMID- 12562219 TI - Assessment of abdominal pain in school-age children. AB - Pediatric abdominal pain can be a difficult condition to accurately assess for the nurse to determine whether the child's need is for teaching, treating, or transferring. This article describes the process as well as practical tips to be used by the nurse in the school setting. Distinguishing characteristics and findings, including key physical assessments, of some of the most frequent or most important conditions are described. Atypical, but not to be overlooked, causes are also discussed. PMID- 12562220 TI - Developing a nursing protocol for over-the-counter medications in high school. AB - Management of medications in school is one of the critical roles that school nurses carry out in the school setting. In recent years, parents have come to question the medication procedures that school districts follow. Parents question why a physician's order is required for school personnel to provide over-the counter (OTC) medications to their child at school. How do school districts balance the safety of students with the needs of parents wanting their children to have access to OTC medications at school? Following legal guidelines helps to reduce the risk for school nurses. Through the development and utilization of Nursing Standardized Protocols, high school nurses are able to provide nonprescription analgesics for specific common student complaints such as noninjury headaches and dysmenorrhea. On the basis of nursing knowledge and judgment, school nurses provide this service, which results in students returning to class quickly, feeling better, and being ready to learn. PMID- 12562221 TI - Data speak: influencing school health policy through research. AB - School nurses occupy a unique position in relation to school health policy. In addition to facing the demands of promoting and maintaining the health of students, they collect the information that is used to document the implementation of school health policy. Effective school health policy is guided by reliable, credible data regarding what interventions achieve desired results. Research data that truly reflect school nurses' contributions to student health and educational outcomes enhance the visibility of professional school nurses. School nurses can learn to use data as a policy tool by knowing data collection methods, when to select a particular method, and how outcomes research can benefit school nurse practice. PMID- 12562222 TI - A mentoring program for new school nurses. AB - Until recent years, school nursing practice consisted mainly of screenings and first aid. However, the changing health, social, and emotional needs of children in the school setting have brought about an expansion of school nursing services. Now school nurses must not only perform routine first aid and screenings, but they must also carry out independent health assessments, monitor various medications, counsel students from troubled homes, collaborate with their educational partners, and complete numerous reports. Historically, school nurses began their careers with little specialized education or orientation. However, with the increased demands in school nursing, it is important to guide, advise, and mentor new school nurses so they can reach a level of competency and confidence more quickly. In this descriptive study, new school nurses were asked to indicate their level of satisfaction with the School Nurse-Mentoring Program. The findings of this study indicate that a mentoring program for new school nurses results in increased career satisfaction, a higher level of competency, and health benefits for the students being served. PMID- 12562224 TI - Parents' perspectives on homelessness and its effects on the educational development of their children. AB - This qualitative study explored parents' perceptions of how their homelessness affected the development and academic achievement of their children. Grounded theory with symbolic interactionism was the framework for this study. Data were collected through semistructured interviews with 34 homeless families in a variety of settings. Multiple factors were found, including unstable relationships, abuse and violence, abdication of parental responsibility, poor parenting models, and resilient children. The findings present a case for supportive educational services for homeless school-age children. School nurses play a dual role. They can ensure that school personnel and resource providers understand the culture of homelessness, and they can develop and implement innovative programs for parents and school personnel to help homeless children. PMID- 12562223 TI - Anger assessment in rural high school students. AB - Anger and aggression in school children are a major concern in American society today. Students with high anger levels and poor cognitive processing skills are at risk for poor relationships, underachievement in school, and health problems. This article describes characteristics of children who are at risk for high anger levels and aggression as well as those who are able to modulate their anger. Results of a survey are reported which describe levels of anger in 624 rural high school students. This sample reported lower levels of anger, compared to the normative group. Differences among the sample include higher internal anger expression in girls, higher trait anger in boys ages 15-16, and higher trait anger in girls ages 16-17. Recommendations are made for future research and specific steps that school nurses can take. PMID- 12562225 TI - Nursing discipline: demystifying the process. AB - This article provides a general overview of the disciplinary process for nurses. It also outlines at least one example of an alternative, often called a diversion program, to the standard disciplinary process. It urges school nurses to obtain legal counsel when responding to an inquiry from the board of nursing and discusses several factors to consider when seeking an attorney. The article also briefly discusses how the Nurse Licensure Compact affects the disciplinary process. PMID- 12562226 TI - The detection of precancerous cervical lesions can be significantly increased. PMID- 12562228 TI - Six-year trends in laboratory computer availability. AB - CONTEXT: Failure of a clinical laboratory computer system can disrupt work flow and charge capture and affect patient care. The first comprehensive survey of computer downtime was conducted in 1995 and demonstrated significant interinstitutional variation in system availability. Despite numerous changes in the laboratory and computer industries since 1995, no follow-up study has been reported. OBJECTIVES: To quantify current laboratory computer availability and compare it with 1995 performance. DESIGN: Ninety-seven laboratories prospectively recorded the frequency and duration of computer downtime during 30 days in 2001. Results were compared with 1995 survey data. RESULTS: For the median facility, the number of downtime episodes decreased from 8 events per 30 days during 1995 to 3 events per 30 days during 2001 (P <.01). The frequency of unscheduled downtime also improved, from a median of 2 to 0.5 events per 30 days (P <.01). Reduced downtime events were paralleled by reduced cumulative downtime (14.3 vs 4.0 hours per 30 days; P <.01). Improvements were not restricted to the median facility; laboratories performing in the bottom quartile in 2001 recorded substantially less downtime than laboratories in the bottom quartile in 1995. When the comparison was restricted to the 37 institutions that participated in both the 1995 and 2001 surveys, a significant reduction in overall downtime and unscheduled downtime events was still evident (P <.01). More recent installation of vendor software patches was associated with a reduced frequency of downtime events in the 2001 data set. CONCLUSION: Laboratory computer downtime was less frequent in 2001 than in 1995; industry performance appears to be improving. PMID- 12562227 TI - Multilaboratory evaluation of serum analysis for HLA antibody and crossmatch reactivity by lymphocytotoxicity methods. AB - CONTEXT: This report presents results of the serum antibody analysis and crossmatch challenges in the proficiency testing program for histocompatibility testing jointly sponsored by the American Society for Histocompatibility and Immunogenetics and the College of American Pathologists. OBJECTIVE: To obtain information about consensus rates among participating laboratories that reported antibody screening and crossmatch results by direct complement-dependent lymphocytotoxicity (CDC) and/or anti-human globulin (AHG)-augmentation methods. DESIGN: We analyzed responses from approximately 165 laboratories participating in 32 surveys during 1993-2000. Most of the testing was done by CDC methods, but increasing proportions of laboratories are using AHG augmentation of these techniques; almost one half of the serum screenings and crossmatches were done by AHG. RESULTS: A total of 40 serum specimens were screened to determine the percent panel-reactive antibody (PRA) and identify HLA-specific antibodies. Participants often reported very wide ranges of PRA values. Panel-reactive antibody ranges exceeded 60 percentage points for 16 (40%) of the serum screening results by CDC and for 31 (77%) of the results by AHG. The interlaboratory variability of PRA values suggests that in many laboratories, the CDC or AHG procedures were often too insensitive or overly sensitive. The antibody identification results revealed inconsistent patterns among the participants performing CDC or AHG screening. Most participants reported the same primary antibody specificities by both methods. The consensus levels were generally high for the monospecific sera. On the other hand, there was much less agreement among the participants if the sera reacted with 2 or more HLA antigens. Participants using the more sensitive AHG method reported additional antibody specificities in many specimens, but invariably the consensus levels were rather low. A total of 192 serum-cell combinations were used for the crossmatch challenges. There was considerable interlaboratory variability; 21% of the CDC crossmatches and 36% of AHG crossmatches failed to reach the 90% consensus threshold. CONCLUSIONS: This experience demonstrates considerable inconsistencies in serum screening and crossmatching among laboratories participating in the American Society for Histocompatibility and Immunogenetics/College of American Pathologists surveys. A lack of uniformity in test results may limit the efficient application of these methods in a clinical setting. Standardization of crossmatch and antibody screening techniques is highly desirable. PMID- 12562229 TI - Specimen collection volumes for laboratory tests. AB - CONTEXT: Unnecessary tests, inefficient ordering practices, and collection of more blood than is required for testing contribute to iatrogenic anemia in hospitalized patients. Laboratories accredited by the College of American Pathologists are expected to review phlebotomy practices for specimen collection volumes periodically. OBJECTIVE: To report specimen collection, analytic, and discard volumes for routine laboratory tests and to identify practice variables associated with overcollection and blood wastage. DESIGN: Clinical laboratories participating in the College of American Pathologists Q-Probes laboratory improvement program recorded collection container size, laboratory-defined requested volume, manufacturer-defined analytic volume, and average discard volume for routine complete blood cell counts and electrolyte panels ordered for patients in intensive care units. Participants provided information about their specimen collection, processing, and analytic practices in a questionnaire. SETTING AND PARTICIPANTS: A total of 140 public and private institutions. MAIN OUTCOME MEASURES: Overcollections for routine collections and for situations in which a reduced volume of specimen is collected, and average discard volume per tube. RESULTS: Laboratories collected a median of 2.76 mL (or 8.5 times) more than their instrument's analytic volume for routine complete blood cell counts and 1.75 mL (or 12 times) more than their instrument's analytic volume for routine electrolyte panels. For clinical situations in which reduced collection volumes were necessary, overcollection for the same analytes was 0.5 mL (3 times) and 0.44 mL (4.2 times), respectively. The median discard volume was 2.8 mL/tube for complete blood cell counts and 2.0 mL/tube for electrolyte panels. Specimen collection container size was directly associated with overcollections and discard volumes. Instrument analytic volume was not a determinant of blood wastage. CONCLUSIONS: Most laboratories can decrease collection volumes without compromising the ability of the laboratory to report a reliable and timely result. Use of smaller collection tubes can help reduce blood wastage. PMID- 12562230 TI - Age and laboratory costs for hospitalized medical patients. AB - OBJECTIVE: To examine the hypothesis that older hospitalized patients have higher laboratory costs than younger patients in the same severity-adjusted diagnosis related group (DRG). DESIGN: We obtained hospital case mix data sets (1995-1997) from the Massachusetts Division of Health Care Finance and Policy. We selected discharge abstracts from 4 medical DRGs, at 5 large academic hospitals (n = 15,265) and 5 midsized community hospitals (n = 10,540), for analysis. We converted laboratory and blood product charges to direct costs using the department-specific ratio of cost to charges. We adjusted diagnostic groups for severity of comorbid conditions and complications using the refined DRG method. MAIN OUTCOME MEASURES: Hospital length of stay (LOS), laboratory direct cost (LDC) per hospitalization, LDC per hospital day, and ratio of LDC to total direct cost. RESULTS: Hospital LOS was longer for older patients in all comparisons. Laboratory direct cost per hospitalization was higher for older patients in some DRGs, but lower in other DRGs. Laboratory direct cost per hospital day was almost always less for older patients than for younger patients, both at academic and community hospitals. Data stratification by gender, admission status, and principal diagnosis yielded substantially the same pattern of cost differences observed within the larger data set. CONCLUSIONS: Older medical patients have longer hospital stays and generally higher costs. These patients also have a significantly decreased rate of laboratory resource consumption over the course of hospitalization (LDC per hospital day), as well as lower laboratory costs as a proportion of total costs. Age-specific differences in LOS and cost parameters were essentially unchanged after controlling for several potential sources of bias. PMID- 12562232 TI - Clinical-histopathologic concordance of tumors of the nervous system at the Manuel Velasco Suarez National Institute of Neurology and Neurosurgery in Mexico City. AB - CONTEXT: When making a diagnosis, the main purpose of clinicians should not be to achieve certainty, but to decrease diagnostic uncertainty in order to make optimal therapeutic decisions. Diagnostic concordance is an essential characteristic if a measurement is to be considered scientific. In the case of tumors of the nervous system (TNS), one of the most accurate diagnostic tests is magnetic resonance imaging. However, histopathologic analyses are essential, because they refine the diagnosis, benefit the patient, and improve our understanding of the disease. By determining the clinical-histopathologic correlation of TNS in one of the main neurologic centers in Mexico, we sought to project reliable morbidity and/or mortality statistics. OBJECTIVE: To assess clinical and histopathologic diagnostic agreement in cases involving TNS admitted to the Manuel Velasco Suarez National Institute of Neurology and Neurosurgery between 1990 and 1999. DESIGN: Cross-sectional diagnostic concordance study, including all clinical hospital records of patients with histopathologically diagnosed TNS, classified according to World Health Organization criteria. RESULTS: Among 2041 TNS cases, the 3 most frequent types were those affecting the neuroepithelial tissue (32.9%), tumors of the sellar region (29.2%), and tumors of the meninges (25.6%). We found that, overall, clinical-histopathologic concordance for these 3 categories was substantial and statistically significant. CONCLUSIONS: Tumors of the nervous system constitute a heterogeneous group of neoplasms. In the present study, clinical diagnoses substantially agreed with pathologic diagnoses. The a priori clinical diagnosis allowed prompt treatment even before diagnostic confirmation by histopathologic analysis, which is the best way to confirm, clarify, and correct a diagnosis. PMID- 12562233 TI - Incidence and significance of cytoplasmic thyroid transcription factor-1 immunoreactivity. AB - CONTEXT: The immunohistochemical identification of thyroid transcription factor-1 (TTF-1) is regarded as the presence of a nuclear pattern of staining and is used to identify tumors of thyroid or pulmonary origin. Although there have been reports of cytoplasmic expression of TTF-1, the significance of this pattern has not been studied in detail. OBJECTIVES: To determine the incidence at which cytoplasmic immunostaining for TTF-1 occurs and to analyze the diagnostic value of this pattern of immunoreactivity. DESIGN: Histologic sections of 361 consecutive cases of neoplasms stained for TTF-1 were reviewed, and those showing cytoplasmic staining in the tumor cells or in nonneoplastic cells were selected. Clinical correlation was obtained on the latter cases regarding the origin of the tumor. SETTING: An immunohistochemistry laboratory in a tertiary-care institution. RESULTS: The 361 tumors were obtained from 29 organ sites and corresponded to primary and metastatic neoplasms. Twenty-three (6.3%) tumors showed cytoplasmic staining for TTF-1. In 13 of these, the primary site of origin was established with certainty: 7 were lung carcinomas (3 primary lung adenocarcinomas, 1 primary large cell carcinoma, 1 metastatic small cell carcinoma to the liver, 1 metastatic adenocarcinoma to a neck lymph node, and 1 metastatic adenocarcinoma to thigh soft tissue), 3 colonic adenocarcinomas (2 metastases to vertebrae and 1 to lung), 1 metastatic breast ductal adenocarcinoma to femur, 1 metastatic laryngeal squamous cell carcinoma to liver, and 1 meningioma involving the orbit bone. There were 3 lung carcinomas with concomitant nuclear immunostaining. Nonneoplastic liver tissue was the most frequent nontumoral tissue in which the cytoplasm stained with TTF-1 antibody. CONCLUSION: Occasional cytoplasmic staining for TTF-1 in tumors is seen, but it is a nonspecific finding; when present, it should be disregarded for diagnostic purposes. PMID- 12562231 TI - Innate immune-related receptors in normal and psoriatic skin. AB - CONTEXT: A precise role for the innate immune system in psoriasis remains to be determined. Surface receptors, including Toll-like receptors (TLRs) that recognize bacterial ligands and CD91, which recognizes heat shock proteins (HSPs), are implicated in both innate and adaptive immunity. OBJECTIVE: Since skin is exposed to various exogenous stimuli, which can provoke or exacerbate psoriasis, we characterized expression and function of TLRs, CD91, and HSPs in normal and psoriatic skin. DESIGN: A variety of skin-derived cells and blood derived cells were analyzed both in vivo and in vitro; samples were obtained from 24 different individuals for innate immune-related receptor expression and function. By comparing and contrasting individuals with healthy skin and psoriatic patients, several specific differences were identified. RESULTS: Immunohistochemistry-based expression profiling revealed TLR1 expression in epidermal dendritic cells (DCs) and dermal dendritic cells (DDCs) in normal skin, as well as in pre-psoriatic skin and psoriatic plaques, with enhanced basal layer keratinocyte (KC) expression in pre-psoriatic and psoriatic plaques compared with normal skin; TLR2 expression primarily by DDCs; and TLR4 expression by epidermal DCs and DDCs, with mid-epidermal-layer KCs displaying cell surface staining. No TLR9 or CD14 was detected on DCs or KCs, although psoriatic plaques contained CD14-positive macrophages. Analysis of psoriatic epidermis revealed HSPs 27, 60, and 70. Keratinocytes were CD91 negative, but CD91 was expressed by fibroblasts and DDCs in normal and pre-psoriatic skin, with prominent accumulation of CD91 positive DDCs in psoriatic plaques. Cultured KCs revealed no surface expression of TLR2, TLR4, TLR9, or CD91. Exposure of fibroblasts, but not KCs, to lipopolysaccharide or HSPs triggered nuclear factor (NF)-kappaB activation. Heat shock proteins did induce maturation of blood-derived DCs accompanied by increased interleukin-12 production and enhanced antigen-presenting function. CONCLUSIONS: These data demonstrate distinctive patterns of innate immune-related receptors by specific subsets of cells in normal and psoriatic skin, suggesting functional roles for HSPs and DCs in psoriasis. PMID- 12562234 TI - Low nuclear proliferative activity is associated with nonmetastatic islet cell tumors. AB - CONTEXT: Traditional morphologic features of tumor aggression are of limited value in predicting the malignant behavior of endocrine neoplasms. We explored the potential value of nuclear proliferative activity (using Ki-67 immunostaining with semiquantitative scoring) in predicting the clinical behavior of pancreatic islet cell tumors (ICTs), and we correlated this characteristic with hormone expression. OBJECTIVE: To evaluate whether Ki-67 immunostaining using a semiquantitative scoring system has value in predicting the clinical behavior of pancreatic ICTs. DESIGN: We studied 39 pancreatic ICTs from 39 patients. Twenty two ICTs did not metastasize in a median follow-up period of 91 months. The remaining 17 neoplasms did produce metastases (8 in liver, 7 in regional lymph nodes, and 2 in peritoneum). Immunohistochemistry was performed using antibodies to Ki-67 and pancreatic hormones (insulin, glucagon, gastrin, somatostatin, pancreatic polypeptide, vasoactive intestinal polypeptide, and corticotropin). A semiquantitative Ki-67 grading system was followed. The nuclear proliferative activity, as determined by a positive reaction for Ki-67, was considered low (<5% of cells staining positively), intermediate (5%-25% of cells staining positively), or high (>25% of cells staining positively). RESULTS: The majority of the nonmetastatic ICTs (16 cases, 73%) demonstrated either negative or low staining for Ki-67 (P <.001). Conversely, all metastatic ICTs expressed at least an intermediate-grade reaction. High nuclear proliferative activity was only seen in metastatic neoplasms (3 cases, 17%). There was no relationship between immunoexpression of pancreatic hormones and nuclear proliferative activity by either group of tumors. CONCLUSION: An ICT with low nuclear proliferative activity is unlikely to metastasize, whereas high proliferative activity is associated with a metastatic phenotype. Immunohistochemical assessment of Ki-67 using a semiquantitative scoring system is a simple and reliable detection method of cellular proliferative activity in ICTs of the pancreas. PMID- 12562235 TI - Comparative analysis of conventional Papanicolaou tests and a fluid-based thin layer method. AB - CONTEXT: A fluid-based, direct-to-vial method of thin-layer gynecologic cytology (ThinPrep Pap Test) is reported to be more effective than the conventional Papanicolaou test in the detection of squamous intraepithelial lesions. OBJECTIVE: This retrospective analysis evaluated the validity of the findings on the thin-layer method using case material at a large independent laboratory and represented a comparison of performance of both methods over an identical period. METHODS: Data for conventional and ThinPrep tests were compared for 2 periods. Period 1 included 1,421,080 conventional and 56,835 ThinPrep specimens, and period 2 included 564,270 conventional and 109,784 ThinPrep specimens. Squamous intraepithelial lesions were used to determine detection of disease. These 2 sets of data were also analyzed to eliminate effects of any selection bias toward ThinPrep for high-risk patients. RESULTS: Use of ThinPrep showed a greater than 100% increase in the detection rate of squamous intraepithelial lesions (1.3% 3.4% in period 1 and 1.3%-2.9% in period 2), which was statistically significant after correcting for selection bias. We also found a significant decrease in the false-negative proportion (57% in period 1 and 35% in period 2). There was a marked improvement (233%) in the detection of high-grade squamous intraepithelial lesions in high-risk cases and a decrease in the atypical squamous cells of undetermined significance to squamous intraepithelial lesion ratio from 3.1 to 1.5 in period 2. CONCLUSION: ThinPrep is better than the conventional Papanicolaou test in detecting squamous intraepithelial lesions and is a superior screening test in detection of precancerous changes of the cervix. PMID- 12562236 TI - Conformity to guidelines for pathologic examination of the placenta. AB - CONTEXT: The College of American Pathologists developed a consensual guideline for placental examination that included indications for the submission of placentas for pathologic examination. The adherence to this guideline is not known. OBJECTIVES: To identify the number of placentas that were and that should have been examined by a tertiary-care hospital according to the College of American Pathologists' practice guideline and to compare the indications listed by medical staff on their pathology request forms with the clinical events recorded on the hospital's databases. DESIGN: Data from the hospital computer databases and from pathology request forms were collected for all 987 deliveries occurring at a tertiary-level maternity hospital from April through June 2000. RESULTS: Fewer than 20% of placentas were examined, but about 50% should have been. Maternal fever and suspected neonatal infection were the indications with the lowest examination rates. Neonatal indications were infrequently listed. CONCLUSIONS: This hospital examined approximately one third of the placentas that should have been examined. When the placentas were examined, the medical staff often failed to appropriately list the indications on their pathology request forms. PMID- 12562238 TI - A microdissection and molecular genotyping assay to confirm the identity of tissue floaters in paraffin-embedded tissue blocks. AB - CONTEXT: A recurring problem in surgical pathology practice is specimen mix-up and floater contamination. While many cases can be resolved histologically, a significant number remain unclear and may have serious clinical and medicolegal implications. OBJECTIVES: To design a microdissection and genotyping assay to identify contaminating floater tissues in paraffin-embedded tissues that is optimized for small samples, and to use the assay to resolve a series of clinical cases with floater tissues. MATERIALS AND METHODS: Twenty-one cases of possible tissue floater contamination in paraffin-embedded tissue blocks were included. Using 4 unstained, 4-microm-thick histologic sections, multiple sites were microdissected under direct visualization either by hand or by laser capture microdissection. Nonneoplastic and neoplastic tissues were sampled. Polymerase chain reaction was performed for a panel of 10 polymorphic microsatellite markers at 1p34, 3p26, 5q21, 9p21, 10q23, and 17p13. Allele size and content were analyzed semiquantitatively by fluorescent capillary electrophoresis, and the genotypes for the tissues in the paraffin-embedded tissue blocks were compared for identity. RESULTS: Tissue identification was successful in all cases, despite small tissue sample size and fixation effects. Comparative analysis of neoplastic tissue floaters and the presumptive source tumor was performed when possible to control for possible allelic loss or microsatellite instability. CONCLUSIONS: Microdissection and genotyping are effective and reliable means to objectively resolve problems of possible floater contamination. Even minute tissue samples provide sufficient DNA template for polymerase chain reaction microsatellite analysis. Because of the potential clinical implications of floaters, we recommend that all suspected floaters that would change a diagnosis from benign to malignant be subjected to genotyping assay to confirm the identity of the floater tissue. PMID- 12562237 TI - Expression of p53, c-Myc, or Bcl-6 suggests a poor prognosis in primary central nervous system diffuse large B-cell lymphoma among immunocompetent individuals. AB - CONTEXT: Primary central nervous system (CNS) diffuse large B-cell lymphoma (DLBCL) in immunocompetent individuals, although rare, has been rising in incidence. Currently, no reliable prognostic markers are available for these individuals. OBJECTIVE: To study the implications of expression of a panel of oncogenic proteins (Bcl-2, Bcl-6, and c-Myc) and p53 for predicting clinical outcome, particularly overall survival, in immunocompetent individuals with primary CNS DLBCL. DESIGN: Fourteen primary CNS DLBCL cases were retrospectively studied by immunohistochemistry on formalin-fixed, paraffin-embedded sections for the expression of c-Myc, Bcl-2, Bcl-6, and p53. RESULTS: The overall frequencies of expression for p53, c-Myc, Bcl-2, and Bcl-6 in these cases were 29%, 50%, 71%, and 57%, respectively. Cases with expression of p53, c-Myc, or Bcl-6 had a poorer overall survival than those without (Kaplan-Meier survival analysis: 50% cumulative overall survival, 2 months vs 30-60 months, P =.02, log-rank test; 9 16 months vs 21-60 months, P =.03, log-rank test; and 9-16 months vs 21-60 months, P =.16, log-rank test, respectively). The expression of Bcl-2 or proliferation activity by MIB-1 showed no correlation with overall survival. Likewise, the clinical parameters, including age, location of tumors, multiplicity of tumor lesions, and lactase dehydrogenase levels revealed no impact on overall survival. CONCLUSION: Our results suggest that patients with expression of p53, c-Myc, or Bcl-6 have a poorer overall survival than those without. Since traditional prognostic markers in non-CNS DLBCL, such as staging and International Prognostic Index scores, are not applicable to primary CNS DLBCL, evaluation of p53, c-Myc, and Bcl-6 by immunohistochemistry may be warranted as part of prognostic evaluation in immunocompetent patients with primary CNS DLBCL. Further studies are indicated to confirm our observations. PMID- 12562239 TI - Multiple Langerhans cell nodules in an incidental thymectomy. AB - The isolated appearance of Langerhans cell histiocytosis in the thymus is quite rare. We report the finding of multiple small Langerhans cell nodules in the thymus of an 11-month-old infant who had undergone thymectomy in the course of surgical correction of tetralogy of Fallot. The subtle appearance of some of these nodules and aggregates was unlike the pattern of massive thymic infiltration seen in 7 previously reported cases of thymic Langerhans cell histiocytosis in childhood. This difference led us to consider whether these aggregates might be common in the thymus but overlooked in thymus glands removed incidental to cardiac surgery. We examined histologic material from 22 sequential thymectomy specimens removed during cardiac surgery from infants and children. None (0/22) had cohesive aggregates of Langerhans cells, as were seen in the index patient's thymus. Our patient's small thymic nodules resemble more closely a pattern found in adult patients with myasthenia gravis who have been incidentally shown to have Langerhans cell histiocytosis. PMID- 12562240 TI - Large cell neuroendocrine carcinoma of the ampulla of vater. AB - We report a large cell neuroendocrine carcinoma arising in the ampulla of Vater. The patient, a 74-year-old woman, presented with a 3-cm ulcerated mass located in the ampullary region. She died of disease 8 months after surgery. Microscopically, the tumor was extensively necrotic. It was composed of islands and trabeculae irregularly infiltrating the muscular wall of the duodenum. Neoplastic cells were large and had a high mitotic index. Immunohistochemically, they expressed cytokeratin, chromogranin, synaptophysin, and neuron-specific enolase. Large cell neuroendocrine carcinoma is very rare in the ampulla of Vater, and it shares with its more common pulmonary counterpart the same morphology and probably the same poor prognosis. PMID- 12562241 TI - Nocardiosis with brain abscess due to an unusual species, Nocardia transvalensis. AB - The identification of Nocardia transvalensis, an unusual and probably underrecognized cause of nocardial infection, is clinically significant because of this species' resistance to aminoglycosides, a standard antinocardial therapy. Diagnosis requires analytic methods available predominately in reference laboratories. We report a case of disseminated infection with N transvalensis with primary pulmonary involvement and subsequent development of brain abscesses, and review the literature to date. Familiarity with the epidemiology, pathologic findings, and clinical significance of this and other unusual Nocardia species may increase early identification and antibiotic susceptibility testing in cases of nocardial infection. PMID- 12562242 TI - A case of myxoid adrenocortical carcinoma with extensive lipomatous metaplasia. AB - We report a combination of unusual myxoid change and extensive lipomatous metaplasia of an adrenocortical carcinoma. The patient was a 38-year-old man with hypertension and heart failure. Radiographic examination revealed the presence of a left adrenal tumor, and adrenalectomy was performed. The tumor weighed 380 g and appeared encapsulated. The cut surface was predominantly gelatinous. Histologically, the tumor was composed of atypical round cells with eosinophilic to vacuolated cytoplasm. The tumor was diagnosed as adrenocortical carcinoma. The stroma accumulated copious mucinous material. In addition, individual to nodular mature adipocytes were admixed throughout the tumor. The transition from carcinoma cells to mature adipocytes was recognized. Myxoid change is a very rare phenomenon in adrenocortical carcinoma, and only 10 similar cases have been reported to date. Lipomatous metaplasia is another peculiar feature of adrenocortical lesions that has been reported only in benign conditions. To our knowledge, this is the first reported case of adrenocortical carcinoma with lipomatous metaplasia. PMID- 12562243 TI - Herpes simplex pancreatitis. AB - Lesions of the pancreas induced by viral infection have drawn relatively little attention because of their low incidence, and the histopathologic features of viral pancreatitis have not been fully elucidated. We report the autopsy findings of 2 patients, a 59-year-old woman with allergic granulomatous angiitis and a 73 year-old man with invasive pulmonary aspergillosis who had a disseminated visceral herpes simplex virus (HSV) infection. In both cases, the liver was the organ most severely affected by the viral infection. The pancreas showed multiple small foci of hemorrhagic necrosis, which were not accompanied by fat necrosis of the surrounding adipose tissue. Histopathologically, Cowdry type A intranuclear inclusions and a ground-glass appearance of the nuclei were found in many degenerated acinar cells around the necrotic foci. The gross appearance and histopathologic features of HSV pancreatitis were characteristic and, in particular, distinct from those of the more common acute hemorrhagic pancreatitis. Immunohistochemistry using an anti-HSV antibody revealed immunoreactivity in the intranuclear inclusions and ground-glass nuclei, and polymerase chain reaction analysis disclosed that the causative virus in these 2 cases was HSV-1. Herpes simplex virus pancreatitis constitutes a rare, but distinct pathologic entity among a group of acute pancreatitis diseases with diverse etiopathogenesis. PMID- 12562244 TI - Human parainfluenza virus giant cell pneumonia following cord blood transplant associated with pulmonary alveolar proteinosis. AB - Giant cell pneumonia secondary to human parainfluenza virus 3 has been reported only rarely in immunocompromised hosts. The few cases documented after bone marrow transplant have resulted in significant morbidity and mortality. To our knowledge, this entity has not been described following umbilical cord blood transplant. Pulmonary alveolar proteinosis, a rare condition that has been reported with increasing frequency in association with immunocompromise and infections, has not been documented in the setting of either umbilical cord blood transplant or human parainfluenza viral infection. We report what we believe is the first documented case of giant cell pneumonia caused by human parainfluenza virus 3 in an umbilical cord blood transplant recipient. To our knowledge, a unique associated feature of this case, a pulmonary alveolar proteinosis-like reaction, has not been reported previously in association with human parainfluenza virus pneumonia. PMID- 12562245 TI - Pathologic quiz case: a scrotal sac mass incidentally discovered during autopsy. PMID- 12562246 TI - Pathologic quiz case: clear cell tumor of the pancreas associated with hypercalcemia. PMID- 12562247 TI - Pathologic quiz case: a patient with acquired immunodeficiency syndrome and an unusual biliary infection. PMID- 12562248 TI - Pathologic quiz case: a renal tumor in a 62-year-old woman. PMID- 12562249 TI - Pathologic quiz case: refractory hiccups in a man after liver transplantation for hepatitis C. PMID- 12562250 TI - Bilateral intrathyroidal lymphoepithelial cysts. PMID- 12562251 TI - Hairy cell leukemia with marked lymphocytosis. PMID- 12562253 TI - Hereditary nonpolyposis colorectal cancer syndrome in a patient with urothelial carcinoma of the upper urothelial tract. AB - Urothelial carcinoma of the upper urinary tract is relatively uncommon but may develop as a manifestation of the hereditary nonpolyposis colorectal cancer syndrome (HNPCC), which is characterized by mutations in a number of DNA mismatch repair genes and detectable as microsatellite instability or loss of the respective protein by immunostaining. No well-established screening test is available for urothelial carcinomas of the upper urinary tract, and little is known of the clinical impact of screening for HNPCC in patients with upper urinary tract cancer. We describe herein a patient with a urothelial carcinoma of the ureter and a strongly positive history of cancer, who was subsequently found to have HNPCC. Our findings reinforce the importance of obtaining a comprehensive history of cancer in patients with urothelial carcinoma of the renal pelvis and ureter. Subsequent identification of individuals with HNPCC enables the patient and at-risk relatives to benefit from targeted surveillance and management programs. PMID- 12562254 TI - Synchronous adenocarcinoma and mantle cell lymphoma of the colon. AB - Synchronous occurrence of malignant lymphoma and carcinoma, both located in the intestinal tract, is unusual. We report a unique case of an adenocarcinoma of the cecum and a simultaneous mantle cell lymphoma of the colon, terminal ileum, and regional lymph nodes in an 85-year-old man. Grossly, the adenocarcinoma was identified as a cecal mass. Lymphomatous involvement of the gastrointestinal tract was evident only on microscopic examination. The terminal ileum and colon showed microscopic disseminated multiple mucosal nodules, with involvement of the regional lymph nodes. There was no involvement of distant organs, suggesting that the mantle cell lymphoma was early in its evolution without formation of polyps or a mass lesion. To our knowledge, this is the fourth reported case with this association and the second case that showed early involvement of the gastrointestinal tract with mantle cell lymphoma without polyp formation. PMID- 12562255 TI - Pulmonary embolization of bovine collagen. AB - A bovine collagen matrix is sometimes used as a delivery medium during direct intratumoral injection of a chemotherapeutic agent. The bovine collagen enhances the dose and duration of local drug delivery and limits systemic toxicity. Although this strategy is advocated as a means of easy and effective delivery of chemotherapeutic drugs, the associated risks are not well defined. We report the case of a 71-year-old man with hepatocellular carcinoma who underwent weekly intratumoral injections of cisplatin in a bovine collagen matrix. During the third injection, he suddenly and unexpectedly underwent cardiac arrest and died. An autopsy disclosed diffuse occlusion of the pulmonary microcirculation by bovine collagen. The collagen emboli were associated with an inflammatory infiltrate typical of bovine collagen-induced hypersensitivity. This case identifies a fatal complication of intratumoral chemotherapy injections using a bovine collagen matrix, which does not appear to have been previously reported. This case underscores the valuable role of the traditional autopsy examination as a means of identifying possible complications of novel oncologic strategies, which are being rapidly developed and implemented. PMID- 12562256 TI - Two cases of epithelioid angiosarcoma involving the thyroid and a brief review of non-Alpine epithelioid angiosarcoma of the thyroid. AB - Epithelioid angiosarcoma involving the thyroid is a rare entity, more often described in the Alpine region. Two cases of epithelioid angiosarcoma that affected the thyroid in patients from a non-Alpine location were diagnosed during a 10-year period in our department. The first case occurred in an 89-year-old Chinese man with a history of longstanding goiter, whereas the second case involved a 74-year-old Chinese man with a history of angiosarcoma of the scalp. On histologic examination, both thyroid tumors were composed of plump epithelioid cells with vesicular chromatin and prominent nucleoli, forming vascular structures and solid sheets. Positive staining for CD31 and factor VIII-related antigen confirmed endothelial differentiation in both cases. Both patients died within 5 months following the diagnosis of thyroid disease. The relationship of the scalp angiosarcoma and thyroid disease of the second patient is unclear. A brief review of non-Alpine primary thyroid epithelioid angiosarcoma is presented. PMID- 12562257 TI - Rosai-Dorfman disease presenting in the gastrointestinal tract. AB - Rosai-Dorfman disease (RDD) rarely presents a diagnostic challenge. However, unusual sites of presentation can cause confusion. We present a case of RDD of the colon in a symptomatic 51-year-old woman. The gastrointestinal tract is the organ system least commonly affected by RDD, with only 11 cases reported. Most cases have been found incidentally or at autopsy. The widespread use of endoscopic procedures makes it more likely that this entity will be encountered. Most patients with gastrointestinal tract involvement have evidence of widespread extranodal disease and have a protracted clinical course. PMID- 12562258 TI - Composite thymoma and chronic lymphocytic leukemia/small lymphocytic lymphoma involving the anterior mediastinum. AB - Simultaneous involvement of the anterior mediastinum by thymoma and B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), to our knowledge, has not been previously reported. We describe a composite tumor composed of thymoma and CLL/SLL incidentally discovered in a 62-year-old man who had no history of malignant diseases or immunologic disorders. The preoperative peripheral blood specimen showed a normal complete blood cell count and differential count. The diagnosis was established by histologic examination and immunophenotypic studies of the surgically excised anterior mediastinal mass. Postoperatively, bone marrow aspiration and biopsy specimens showed morphologic evidence of CLL/SLL, and the presence of neoplastic cells in peripheral blood and bone marrow was confirmed by flow cytometry immunophenotypic analysis. PMID- 12562259 TI - Glomerular calcification in hypercalcemic nephropathy. AB - Hypercalcemic nephropathy has been classified as a tubulointerstitial renal disease. The presence of glomerular pathologic findings attributable to hypercalcemia has been observed in only a few patients and therefore has been considered an unusual finding. In the current study, calcium deposition within glomeruli was investigated in 2 patients with extreme elevations in serum calcium levels and hypercalcemic nephropathy. The study material consisted of a renal biopsy specimen from a 31-year-old woman (patient 1) who had T-cell lymphoma/leukemia and a serum calcium level of 20.2 mg/dL (5.0 mmol/L) and autopsy kidney specimens from a 19-year-old woman (patient 2) who was being evaluated for primary hyperparathyroidism and a calcium level of 18.4 mg/dL (4.6 mmol/L). The renal biopsy specimen for patient 1 exhibited calcium deposits present in the glomerular capillary basement membranes, where they were associated with segmental sclerosing lesions (21% of glomeruli). Nine percent of the cortical tubules contained calcifications. In patient 2, calcium was found in the mesangial areas in 95% of glomeruli, filling the Bowman space in 7% of glomeruli, or associated with capillary basement membranes and segmental sclerosing lesions (12% of glomeruli). Fifteen percent of cortical tubules, 4% of outer medullary tubules, and 40% of inner medullary tubules were calcified. In neither case was there immunofluorescence or electron microscopic evidence of primary glomerular disease. Thus, glomerular calcification may exceed that occurring in the cortical and outer medullary tubules and may play a significant role in the loss of renal function in hypercalcemic nephropathy. Glomerular calcinosis may also be recognized as an additional cause of segmental glomerulosclerosis and nephrotic range proteinuria in patients with extremely high levels of serum calcium. PMID- 12562260 TI - Hidradenoma papilliferum of nasal skin. AB - Pathologic examination of an excisional biopsy specimen of a subepidermal nodule from the nasal tip of a 37-year-old black man demonstrated papillae covered by eosinophilic columnar epithelial cells, suggesting cylindric cell papilloma or well-differentiated papillary adenocarcinoma of nasal mucosa. The patient did not submit to further investigation of the sinonasal tract but sought attention 15 months later because of local recurrence of the lesion. A protuberant, 3-cm, cystic nodule of the subcutis over the left nasal ala and tip was excised completely and found to have characteristics of hidradenoma papilliferum, an apocrine gland neoplasm most commonly occurring in the anogenital skin of white women. This case demonstrates how easily an apocrine neoplasm arising near a mucocutaneous junction can be confused with a mucosal tumor, potentially leading to inappropriate treatment. PMID- 12562261 TI - Renal cell carcinoma with intratumoral calcium oxalate crystal deposition in patients with acquired cystic disease of the kidney. AB - We describe 2 cases of renal cell carcinoma arising in acquired cystic disease of the kidney (ACDK) in patients with end-stage renal disease undergoing hemodialysis for more than 5 years and provide a brief review of the complications of ACDK. In both cases, abundant calcium oxalate crystals were observed within the tumors. Histologically, one of the tumors was a conventional (clear cell) renal cell carcinoma. The other tumor was a bilateral papillary renal cell carcinoma. Both tumors were high-grade carcinomas with extensive oncocytic (acidophilic) features. Also noted within the kidneys were cysts with atypical papillary hyperplasia. The clinicopathologic findings along with review of the literature suggest a relationship between tumor growth and calcium oxalate crystal deposition in patients undergoing hemodialysis with ACDK. PMID- 12562262 TI - Acute leukemia with myeloid, B-, and natural killer cell differentiation. AB - Biphenotypic acute leukemias account for 4% to 8% of all acute leukemias. Most of these leukemias are of myeloid-B-cell or myeloid-T-cell lineage. Acute myeloid natural killer cell leukemia has been recognized recently. We report the first case, to our knowledge, of CD56(+) acute leukemia showing unequivocal myeloid and B-cell differentiation in a 20-year-old woman, whose blast cells were positive for myeloperoxidase, CD13, CD33, CD117, terminal deoxynucleotidyl transferase, CD19, CD20, CD22, CD34, HLA-DR, and CD56 but negative for CD3, CD5, CD7, and CD10. Rare Auer rods were identified in the blast cells. Polymerase chain reaction assays showed rearrangement of immunoglobulin heavy-chain gene and absence of Epstein-Barr virus DNA. We propose that this novel form of multilineage leukemia may represent the neoplastic counterpart of a progenitor that can give rise to myeloid, B, and natural killer cells. PMID- 12562264 TI - Involvement of Meckel diverticulum in Crohn disease associated with pancreatic heterotopia. AB - Although the association of Crohn disease and Meckel diverticulum has been widely reported, the direct involvement of a Meckel diverticulum by Crohn disease is less common and is usually the result of contiguous spread. The presence of heterotopic tissues in an affected diverticulum is rare and most often consists of gastric mucosa. We report herein the presence of Crohn disease within a Meckel diverticulum, with a normal small bowel immediately adjacent to the diverticulum, associated with pancreatic heterotopia. PMID- 12562263 TI - Malignant perivascular epithelioid cell tumor involving the prostate. AB - Perivascular epithelioid cell tumor (PEComa) is a neoplasm chiefly composed of HMB-45-positive epithelioid cells with clear-to-granular cytoplasm and a perivascular distribution. We describe such a tumor involving the prostate and seminal vesicle in a 46-year-old man. The tumor had characteristic histologic features of PEComa. Immunohistochemically, the tumor cells were positive for HMB 45 but negative for epithelial markers, Melan-A, and S100 protein. The tumor behaved in a malignant fashion, and the patient died of the disease 4 years after diagnosis. PMID- 12562265 TI - Pathologic quiz case: a man with chronic otorrhea. PMID- 12562266 TI - Pathologic quiz case: a 5-month-old infant with respiratory distress. PMID- 12562267 TI - Pathologic quiz case: a 58-year-old man with a pancreatic mass. PMID- 12562268 TI - Pathologic quiz case: a 49-year-old woman with a left axillary mass. PMID- 12562269 TI - Pathologic quiz case: an enlarging, firm nodule on the abdominal skin in an otherwise healthy 73-year-old woman. PMID- 12562270 TI - Pathologic quiz case: a paratesticular mass in a 55-year-old man. PMID- 12562271 TI - Pathologic quiz case: a 50-year-old man with progressive worsening of neurological symptoms. PMID- 12562272 TI - The pathologist's intraoperative role during parathyroid surgery. PMID- 12562273 TI - Therapeutic drug monitoring in human immunodeficiency virus/acquired immunodeficiency syndrome. Quo vadis? PMID- 12562274 TI - Practice guidelines on the reporting of smudge cells in the white blood cell differential count. PMID- 12562275 TI - Pathologic quiz case: An 85-year-old woman with a rapidly growing right neck nodule. PMID- 12562276 TI - Pathologic quiz case: A subcutis nodule of the right thoracic wall. PMID- 12562277 TI - Pathologic quiz case: Pulmonary symptoms and lymphadenopathy in a human immunodeficiency virus-infected woman. PMID- 12562278 TI - Pathologic quiz case: A 43-year-old woman with a subcutaneous mass in the upper arm. PMID- 12562279 TI - Bronchus-associated lymphoid tissue lymphoma. PMID- 12562280 TI - Exflagellating Plasmodium vivax in peripheral blood. PMID- 12562281 TI - Adrenoleukodystrophy. PMID- 12562283 TI - Left-sided splenorenal fusion with marked extramedullary hematopoiesis and concurrent lithium toxicity. A case report and review of the literature. AB - Occasionally, heterotopic splenic tissue can occur in the renal fossa secondary to splenosis following splenic trauma or splenectomy. More rarely, it can represent a developmental anomaly secondary to the fusion of splenic and renal tissues. Splenorenal fusion can present as a renal mass, mimicking primary or secondary renal neoplasms on imaging studies, and patients can also present with symptoms of hypersplenism (anemia). We report a case of splenorenal fusion in a 51-year-old woman who initially presented with lithium toxicity, anemia, thrombocytosis, and a large renal mass that mimicked a primary renal neoplasm. The possible embryologic origin of splenorenal fusion, effects of lithium toxicity, and utility of various imaging modalities are discussed. The literature on renal heterotopic splenic tissue is also briefly reviewed. PMID- 12562285 TI - Lymphangitic spread of hepatocellular carcinoma. AB - Pulmonary lymphangitic carcinomatosis is a well-documented phenomenon caused by spread of carcinoma to the pulmonary vasculature and lymphatics, often resulting in respiratory failure and cor pulmonale. It has been described in numerous types of carcinoma, most commonly occurring with carcinomas of the breast and stomach and with choriocarcinoma. We report the case of a patient who presented with increasing shortness of breath and dyspnea on exertion. Autopsy findings revealed diffuse pulmonary vascular spread of a hepatocellular carcinoma to the lungs. To our knowledge, this is the first reported case of lymphangitic spread of a hepatocellular carcinoma causing respiratory compromise (lymphangitic carcinomatosis). PMID- 12562284 TI - Automated analysis of differentiation-induced leukemic cells during all-trans retinoic Acid therapy of acute promyelocytic leukemia. AB - During differentiation-induction therapy of acute promyelocytic leukemia (APL) patients with all-trans retinoic acid (ATRA), a variety of APL-derived bizarre granulocytic cells appear in the peripheral blood. To evaluate the differentiation induction of leukemic cells, we have developed a new scattergram analyzing program with an automated hematology analyzer and compared the data with the flow cytometry measuring the expression of differentiation-associated cell surface antigens, CD11b and CD16. We used the fluorescence intensity and side scatter as parameters of granulocytic maturation in the analysis with the automated hematology analyzer. The analysis of 2 ATRA-treated APL patients and in vitro study using HL-60 cells demonstrated that the levels of fluorescence intensity and side scatter decreased as accompanied with granulocytic maturation, and these changes were parallel with the results of flow cytometry. Our automated scattergram analysis of cell differentiation will contribute to general, objective, and real-time evaluation of differentiation-induction therapy of APL with ATRA. PMID- 12562286 TI - Sclerosing adenosis of the prostate. Histologic features in needle biopsy specimens. AB - Sclerosing adenosis of the prostate is a pseudoneoplastic lesion that can mimic prostate cancer. Because the lesion is more common in the transition zone, which is only rarely sampled in needle biopsy, it is uncommon to see examples of this lesion in biopsy specimens. Because sampling of the transition zone of the prostate is likely to become more frequent, practicing surgical pathologists must be aware of the morphologic features of sclerosing adenosis of the prostate in needle biopsy specimens, in order to avoid misinterpretation of sclerosing adenosis of the prostate, a benign lesion, as prostate adenocarcinoma. We report the morphologic findings of sclerosing adenosis of the prostate in 3 needle biopsy specimens from 2 patients diagnosed as having sclerosing adenosis. We found a combination of histologic (mainly a cellular myxoid stroma and a double cell population of acinar cells) and immunohistochemical features demonstrating a continuous basal cell layer with myoepithelial differentiation to be diagnostic. PMID- 12562287 TI - Pseudothrombocytopenia associated with infectious mononucleosis. AB - A 22-year-old man was hospitalized for assessment of thrombocytopenia and fever. Examination showed that he had infectious mononucleosis and moderately severe thrombocytopenia that was asymptomatic. Examination of blood smears revealed that the thrombocytopenia was caused by the clumping of platelets. We made a diagnosis of ethylenediaminetetraacetic acid-dependent pseudothrombocytopenia after excluding other infectious mononucleosis-related mechanisms of thrombocytopenia. When the patient recovered from infectious mononucleosis 2 months later, his thrombocytopenia improved, and no platelet clumping in peripheral blood smears was noted. Ethylenediaminetetraacetic acid-dependent pseudothrombocytopenia should always be considered as a possible cause of reported low platelet counts, even in patients with infectious mononucleosis and splenomegaly. PMID- 12562288 TI - Favorable outcome after infusion of coagulase-negative staphylococci-contaminated peripheral blood hematopoietic cells for autologous transplantation. AB - Bacterial contamination of peripheral blood hematopoietic cells collected for autologous bone marrow transplantation occurs sporadically. Although transfusion of contaminated hematopoietic cells without adverse clinical sequelae has been reported, detailed guidelines for transfusing cells with contamination are not available. We report a case of autologous hematopoietic cell transplantation that necessitated using multiple aliquots of peripheral blood hematopoietic cells known to be contaminated with coagulase-negative Staphylococcus bacteria. Prophylactic intravenous antibiotic therapy was given with the infusion of contaminated hematopoietic cells. The patient had positive results on a blood culture, but engraftment was successful, and serious adverse effects did not occur. With appropriate microbial identification and prophylactic antibiotic therapy, contaminated hematopoietic products can be safely infused when necessary with a good clinical outcome. PMID- 12562289 TI - Carcinoid tumors of the urinary bladder. Immunohistochemical study of 2 cases and review of the literature. AB - We report the clinical, histopathologic, and immunophenotypic features of 2 carcinoid tumors of the urinary bladder and review the literature. Both tumors were located in the bladder neck, presented with hematuria, were small, and appeared to be completely excised cystoscopically. The tumors were smooth surfaced sessile polypoid nodules covered by urothelium. Both tumors had glandular architecture. One tumor had regular bland nuclei, and the other had focal moderate nuclear pleomorphism. Neuroendocrine differentiation was readily confirmed by the immunohistochemical markers chromogranin, synaptophysin, and neural cell adhesion molecule (CD56/NCAM). In 1 of the tumors, 20% of the cells were positive for the beta subunit of human chorionic gonadotropin, and 20% were positive for thyroid transcription factor 1. PMID- 12562290 TI - Benign heterotopic epithelial inclusions in axillary lymph nodes. Histological and immunohistochemical patterns. AB - Benign heterotopic epithelial inclusions in axillary lymph nodes are an extremely rare condition that must be differentiated from metastatic carcinoma. We describe 2 histologically different examples of benign epithelial inclusions in nonsentinel axillary lymph nodes, each with an unusual clinical presentation. PMID- 12562291 TI - Primary plasmacytoma arising in an endocervical polyp with detection of neoplastic cells on papanicolaou test. A case report and review of the literature. AB - Primary plasmacytomas of the female genital tract are extremely rare and present a substantial diagnostic challenge. Five cases morphologically representing plasmacytomas and localized to the uterine cervix have been reported previously; however, only 1 was shown to be monotypic for immunoglobulin light-chain expression. We report the case of a 37-year-old woman who had highly atypical plasma cells on her Papanicolaou test. A clinically detected endocervical polyp was removed and revealed a plasmacytoma, the diagnosis of which was substantiated by demonstrating monotypic lambda-light-chain restriction and a clonal immunoglobulin heavy-chain gene rearrangement. The cytologic and histopathologic findings of plasmacytomas of the uterine cervix are discussed, including the utility of immunophenotypic and molecular techniques to confirm the neoplastic diagnosis. PMID- 12562292 TI - Pleomorphic leiomyosarcoma of the adrenal gland. AB - Primary leiomyosarcomas arising in the adrenal gland are exceedingly rare, with only 3 cases reported in the literature. We present the clinical, morphologic, and immunohistochemical features of a pleomorphic leiomyosarcoma, a variant of leiomyosarcoma that has not been described in the adrenal gland. A 63-year-old man presented with a 1-year history of enlarging right upper quadrant mass and pulmonary nodule. A diagnosis of metastatic pulmonary carcinoma to the adrenal gland was rendered on a needle biopsy specimen. Preoperative chemotherapy reduced only the pulmonary mass but not the adrenal mass, which continued to enlarge. Documented by computed tomography and confirmed at surgery, the tumor had completely replaced the right adrenal gland, invading into both the posterior aspect of the right liver and the superior pole of the right kidney. Histologic sections showed a diffuse proliferation of pleomorphic, large, and polygonal neoplastic cells with prominent nucleoli. Many bizarre mitotic figures were present. The neoplastic cells were strongly positive for desmin, calponin, and vimentin. Approximately 80% of the neoplastic cells were positive for the proliferation marker Ki-67. They were negative for smooth muscle actin, muscle specific actin, myoglobin, myogenin, CD117, cytokeratins, carcinoembryonic antigen, epithelial membrane antigen, chromogranin, CD34, CD31, S100 protein, and HMB-45. PMID- 12562293 TI - Molecular evidence of Munchausen syndrome by proxy. AB - Many perpetrators of Munchausen syndrome by proxy present bloodstained materials as counterfeit evidence of proxy hemorrhage. Although blood grouping may show that the blood is not the proxy's, DNA typing may specifically identify the blood's source. A mother claimed that she alone had witnessed gastrointestinal bleeding of her son and presented bloodstained towels as evidence. Several clinical investigations had failed to reveal a bleeding source. I compared the DNA types of the bloodstains and the child's buccal cells. The bloodstain and epithelial cells differed at 4 of 8 microsatellite loci and at the amelogenin locus. The blood and buccal cells shared 1 allele at every locus, suggesting that their sources were closely related. The probability that the source of the blood was maternal was 0.9915 (prior probability, 0.5). I recommend DNA matching in suspected cases of Munchausen syndrome by proxy whenever blood is presented as evidence. PMID- 12562294 TI - Renal cell carcinoma, chromophobe type, with collecting duct carcinoma and sarcomatoid components. AB - We report a case of a 72-year-old man with a chromophobe renal cell carcinoma that had both sarcomatoid and collecting duct carcinoma components. The 7-cm tumor occupied the entire lower pole of the kidney and infiltrated the renal parenchyma and the pelvic-calyceal system. Histologically, it had an area of classic chromophobe renal cell carcinoma that merged into a sarcomatoid component. Closely intermixed with the sarcomatoid component was a collecting duct carcinoma component characterized by highly pleomorphic, epithelioid cells arranged in cords, nests, and tubulomicrocystic structures. The cords, nests, and tubules were associated with a florid desmoplastic stromal response and numerous inflammatory cells. In addition, dysplastic changes were noted in adjacent nonneoplastic collecting duct epithelium. Immunohistochemical studies confirmed the presence of 3 distinct components in this patient's tumor. To the best of our knowledge, this is the first reported case of a chromophobe renal cell carcinoma with sarcomatoid and collecting duct carcinoma components. PMID- 12562295 TI - Pathologic quiz case: A 42-year-old woman with a recurrent right axillary tumor following bilateral mastectomies and axillary resections. PMID- 12562296 TI - Pathologic quiz case: A 23-year-old woman with a polypoid gastric mass. PMID- 12562297 TI - Pathologic quiz case: A cystic mass in the cerebellopontine angle. PMID- 12562298 TI - Pathologic quiz case: An unusual lesion of the mesentery. PMID- 12562299 TI - Pathologic quiz case: A localized pulmonary consolidation in a young woman. PMID- 12562300 TI - Pathologic quiz case: An elderly woman with lymphocytosis. PMID- 12562301 TI - Pathologic quiz case: A 79-year-old woman with an asymptomatic oropharyngeal mass. PMID- 12562302 TI - Pathologic quiz case: Ovarian mass in a 2-year-old girl presenting with pleural effusions. PMID- 12562303 TI - Type II pneumocytes in the evaluation of drug antimycobacterial activity. AB - In order to combat the worldwide increase in the prevalence of multi-drug resistant tuberculosis and Mycobacterium avium complex infections, a number of new antimycobacterial drugs has been synthesised and developed. There is great promise for drugs designed by new strategies, especially those based on the information on mycobacterial genome sequences and a host-parasite relationship. Moreover, the development of new protocols for chemotherapy of intractable mycobacterioses is also needed. For this purpose, better in vitro drug activity assay models that enable prediction of therapeutic activity, particularly those predicting the in vivo sterilising activity, are urgently needed, since the ordinary in vitro methods are inefficient indicators of clinical efficacy. In this context, the in vitro models using Type II pneumocytes, which play an important role in the establishment of mycobacterial pulmonary infections as a portal of mycobacterial organisms to the lungs, are considerably useful, especially in predicting the in vivo activity of certain drugs against Mycobacterium avium complex infection. PMID- 12562304 TI - An evaluation of current cholera treatment. AB - Cholera, caused by Vibrio cholerae O1 and O139, is characterised by profuse purging of watery stools, and vomiting and dehydration. The mainstay of therapy of cholera patients is rehydration with oral rehydration salt solution or intravenous Ringer's lactate depending upon the degree of dehydration. Antibiotics such as tetracycline, doxycycline, norfloxacin, ciprofloxacin and furazolidone may be used as an adjunct to rehydration therapy for severely purging cholera patients to reduce the rate of stool output. This shortens the duration of hospital stay, stops excretion of vibrios in the stool and minimises the requirement of fluids. Resistance to many of these drugs has been observed and is a matter of concern. Other antidiarrhoeals are not recommended. Many antisecretory drugs have been tried as an adjunct therapy, unfortunately, until today, none has been found useful in the treatment of cholera. Feeding during and after cholera is emphasised. PMID- 12562305 TI - Antifungal pharmacotherapy for invasive mould infections. AB - The incidence of invasive mould infections is increasing and is associated with significant morbidity and mortality. Among the most prevalent of these infections are those caused by Aspergillus and Fusarium species. Invasive disease caused by moulds frequently presents as a pulmonary infection, but haematogenous infection can occur. Some moulds cause cutaneous disease through either direct inoculation of the skin or secondary spread to the skin after dissemination from another body site. Early diagnosis can often be difficult and, unfortunately, diagnosis occurs late in the course of illness in many cases. Treatment options have historically been limited by the need for intravenous administration (amphotericin B), significant toxicities (amphotericin B), lack of reliable in vitro activity (e.g., amphotericin B in Fusarium and Scedosporium apiospermum infections) and relative lack of clinical experience with newer agents. The recent approval of voriconazole (Vfend, Pfizer) introduces a treatment option that demonstrates both in vitro and in vivo activity against a variety of moulds. With the recent development of the new echinocandin class of antifungal agents and newer broad spectrum azole antifungal agents with in vitro mould activity, there is a renewed emphasis on fungal treatment strategies. Antimould therapy presents challenges in adverse effect avoidance and management, drug interactions and pharmacoeconomic considerations. Furthermore, combination therapy is being explored with these various new antifungal agents. The administration of an optimal fungicidal therapy early in the course of the illness and control of the underlying disease are vital to prevent complications and mortality from these tenacious mycoses. PMID- 12562306 TI - Antimicrobial therapy of methicillin resistant Staphylococcus aureus infection. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is now one of the commonest causes of nosocomial infection worldwide. The mainstay of treatment until now has been the glycopeptides (vancomycin and teicoplanin). They are not without toxicity and need parenteral administration and monitoring of levels. The increasing frequency of MRSA infections, coupled with the emergence of glycopeptide resistance in S. aureus has made the introduction of new drugs active against Gram-positive organisms essential. New agents active against Gram positive organisms represent either genuinely novel classes of antimicrobials (e.g., oxazolidinones and lipoproteins) or those derived from existing classes (e.g., tetracyclines, glycopeptides, streptogramins and cephalosporins). Some of these newer antibiotics appear to be effective against multi-resistant organisms including MRSA. PMID- 12562307 TI - Preventive strategies of perinatal HIV-1 transmission: an experience from Thailand. AB - There are at least 6 million people in the Asia-Pacific region living with HIV/AIDS. The number of HIV-1 infected women and children is increasing. Important components in order to prevent perinatal HIV-1 transmission include counselling, HIV testing, family planning, obstetric care, antiretroviral agents and avoidance of breast feeding. Zidovudine or nevirapine used as monotherapy or in combination with other antiretroviral agents are recommended as a standard of care in the prevention of perinatal HIV-1 transmission. Each country needs to have its own data concerning HIV/AIDS epidemiology, and the infrastructure and resources to select appropriate strategies in reducing the perinatal HIV-1 transmission rate. PMID- 12562308 TI - Andropause: an androgen deficiency state in the ageing male. AB - An age-related decline in circulating testosterone levels has been shown to occur in the adult male population starting as early as middle age and continuing on into old age. This decline in testosterone has been associated with a number of changes in body composition and sexual performance in the male that are directly attributable to an androgen deficiency state, which can be restored by the use of testosterone replacement therapy. The term 'andropause' has been used to describe this gradual drop in testosterone in the ageing male and is characterised by different endocrine, somatic and psychic changes that become more pronounced as the male gets older. For some males, the adjustment of circulating testosterone levels with replacement therapy to levels seen in young men can improve physical performance, induce a sense of well-being and restore the androgen-dependent sex drive that declines with ageing. PMID- 12562309 TI - Inhaled insulin--a new therapeutic option in the treatment of diabetes mellitus. AB - The beneficial effects of maintaining an optimal glucose control on chronic complications of diabetes have been firmly established in patients with both Type 1 and 2 diabetes mellitus. Effective glucose control usually requires multiple daily injections of subcutaneous insulin. Limitations of these intensive regimens include inconvenience and poor patient adherence and acceptability. As a consequence, several alternative methods for insulin delivery have been developed with the aim of eliminating the pain, inconvenience and disruption of lifestyle associated with the need for insulin injections. Recent evidence suggests that intrapulmonary insulin delivery may be an effective, non-invasive alternative to subcutaneous regular insulin. In fact, clinical trials in the last few years have shown that intrapulmonary insulin was as good as subcutaneous insulin in controlling glucose levels in patients with both Type 1 and 2 diabetes mellitus. It was well-tolerated and, so far, there is no evidence of an increased risk of hypoglycaemia or adverse bronchopulmonary effects. Although the bio-availability of inhaled insulin is lower than that of subcutaneous insulin, the former has a more physiological pharmacokinetic profile than the latter. Inhaled insulin currently represents the most viable alternative insulin delivery method to preprandial subcutaneous injection. PMID- 12562310 TI - Vaginal ring delivery of hormone replacement therapy--a review. AB - Vaginal ring delivery systems are a highly promising approach to hormone replacement therapy (HRT). The vagina is an excellent route for steroid absorption. Menopausal vaginal rings can be designed to allow very low dose local delivery of oestrogen to the vagina for the large percentage of postmenopausal women experiencing urogenital atrophy, or can be modified to deliver much higher doses sufficient for systemic oestrogen replacement therapy, with or without simultaneous progestogen release. Clinical studies of ultra-low dose rings have shown them to be as efficacious as established treatments of urogenital atrophy (creams, pessaries) and to be preferred by most users. Pharmacokinetic and limited clinical studies of vaginal rings designed to deliver systemic doses of HRT (oestrogen alone or combined) have also been highly promising. Vaginal ring delivery of HRT offers women convenient, long-acting therapy (>/= 3 months) that is under their own personal control. PMID- 12562311 TI - Anti-cocaine vaccines: antibody protection against relapse. AB - The past decade has seen the development of several vaccines against illicit drugs. These include vaccines for producing antibodies against cocaine, heroin, methamphetamine and nicotine. The present focus is on anti-cocaine vaccines, as more research has been conducted with these vaccines than other vaccines targeted against a drug of abuse. Attention needs to be given to the structure of the hapten being conjugated, the characteristics of the carrier protein for conjugation with the hapten and the immunisation regimen for antibody production. These issues have an impact on the level of and variability in the anti-cocaine antibodies actively induced and, consequently, on the pharmacokinetic and pharmacodynamic properties of the vaccine. These issues also have an impact on the preclinical and clinical success of the vaccine in protecting against drug use and relapse. If an anti-cocaine vaccine is to be clinically useful, it must induce a sufficient level of antibody in the blood to prevent easy surmountability of protection by continued cocaine use and should be compatible with other treatment medications that may be simultaneously administered. PMID- 12562312 TI - Therapeutic angiogenesis: protein-based therapy for coronary artery disease. AB - Therapeutic angiogenesis is a promising treatment for ischaemic heart disease, particularly for patients who are not candidates for current methods of revascularisation. The goal of angiogenic therapy is the relief of symptoms of coronary artery disease and improvement of cardiac function by increasing perfusion to the ischaemic myocardium. Angiogenic cytokines such as fibroblast growth factor and vascular endothelial growth factor have been studied extensively in preclinical studies. Protein-based therapy with these growth factors has produced functionally significant angiogenesis in several animal models. Enthusiasm following these preclinical results led the way to clinical trials, which so far have shown only modest improvements in myocardial perfusion and clinical outcome. The attenuated angiogenic response to growth factor therapy observed in patients with coronary artery disease may be related to associated conditions such as endothelial dysfunction, regimens of single as opposed to multiple angiogenic agents and inefficiency of current delivery modalities, as illustrated by the disappointing results of the Phase II clinical trials using intravascular techniques of administration. The ultimate role angiogenesis will play clinically in the treatment of ischaemic heart disease will be determined by adequately powered, randomised, double-blind, placebo-controlled trials that include multi-agent angiogenic therapy and intramyocardial methods of delivery. PMID- 12562313 TI - Focal adhesion kinase as a potential target in oncology. AB - Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that plays a pivotal role in signal transduction at integrin-linked cellular adhesions, which mediate cell contact with the extracellular matrix. It has been shown to play a role in the survival of anchorage-dependent cells and to be essential for integrin-linked cell migration - processes that are likely to play important roles in the development of malignancies. FAK is upregulated in a wide variety of human epithelial cancers, with expression being closely correlated to invasive potential. Recently, evidence has emerged directly linking FAK expression to tumour development in vivo, raising the possibility that intervention strategies to block FAK function may potentially provide an opportunity for the development of anticancer therapeutics. PMID- 12562314 TI - Acute pancreatitis--the current status in management. AB - Acute pancreatitis is a life-threatening inflammatory disorder of the pancreas. Currently, there is no effective pharmacological therapy available for this disorder. The management strategies remain supportive. Given the remarkable morbidity and mortality associated with acute pancreatitis, there is clearly a desperate need for effective novel therapies. This paper presents a review on the epidemiology, aetiology, pathogenesis and management of acute pancreatitis and highlights the need for the development of novel and more specific therapies to battle this disorder. PMID- 12562315 TI - Review of lamotrigine and its clinical applications in epilepsy. AB - Lamotrigine is an anti-epileptic agent with broad efficacy. Lamotrigine works at voltage-sensitive sodium channels, thereby stabilising the neuronal membrane and inhibiting the release of excitatory neurotransmitters such as glutamate and aspartate. Early preclinical animal studies indicate its broad-spectrum efficacy, which was later confirmed in clinical trials. Multiple randomised, placebo controlled and comparative trials demonstrate its efficacy against partial and secondarily generalised seizures. Open-label trials show its efficacy against generalised seizures, especially absence seizures of childhood absence epilepsy and generalised seizures of juvenile myoclonic epilepsy. Lamotrigine has a wide clinical dose range and possesses favourable pharmacokinetic properties. It has a good tolerability and safety profile, which enhance compliance. Its small risk of serious skin rash should be weighed against its potential benefits when choosing lamotrigine on an individual basis. Lamotrigine is an excellent therapeutic option in epilepsy. PMID- 12562316 TI - Review of esomeprazole in the treatment of acid disorders. AB - Esomeprazole (Nexium, AstraZeneca) is the (S)-isomer of omeprazole and the first proton pump inhibitor to be developed as an optical isomer. Esomeprazole has an improved pharmacokinetic profile, resulting in increased systemic exposure and less interindividual variability compared with omeprazole, and more effective suppression of gastric acid production compared with other proton pump inhibitors. In several large, double-blind, randomised trials, significantly higher rates of endoscopically-confirmed healing of erosive oesophagitis and resolution of heartburn have been achieved in patients with gastro-oesophageal reflux disease receiving 8 weeks of esomeprazole 40 mg o.d. compared with those receiving omeprazole 20 mg o.d. or lansoprazole 30 mg o.d. In the maintenance of healed erosive oesophagitis, esomeprazole 10, 20 or 40 mg o.d. was significantly more effective than placebo in two 6-month, randomised, double-blind trials. Additionally, esomeprazole 20 mg o.d. was more effective than lansoprazole 15 mg in the maintenance of healed erosive oesophagitis in another 6-month, randomised, double-blind trial. Healing of oesophagitis was also effectively maintained by esomeprazole 40 mg o.d. in a 12-month non-comparative trial. Esomeprazole 20 or 40 mg o.d. effectively relieved heartburn in patients with gastro-oesophageal reflux disease without oesophagitis in two 4-week, placebo-controlled trials. Clinical trials have shown that triple therapy with esomeprazole 40 mg o.d. in combination with amoxicillin and clarithromycin produced Helicobacter pylori eradication rates similar to those obtained using triple therapy involving twice daily dosing with other proton pump inhibitors. Esomeprazole is well-tolerated, with a spectrum and incidence of adverse events similar to those associated with omeprazole. PMID- 12562317 TI - Clinical pharmacology of etoricoxib: a novel selective COX2 inhibitor. AB - The development of COX2 inhibitors with improved biochemical selectivity (such as etoricoxib and valdecoxib) over that of commercially available coxibs has been driven by the potential advantage of safety using higher coxib doses for increased efficacy. Etoricoxib has been approved in the UK as a once-daily medicine for symptomatic relief in the treatment of osteoarthritis (OA), rheumatoid arthritis (RA) and acute gouty arthritis. It is currently approved with additional indications (i.e., for relief of acute pain associated with dental surgery, for primary dysmenorrhoea and for chronic musculo-skeletal pain, including chronic lower-back pain) in Mexico, Brazil and Peru. Etoricoxib has an in vitro COX1/COX2 IC(50) ratio of 344, the highest of any coxib. The administration of therapeutic doses of etoricoxib to healthy subjects does not affect COX1 activity in circulating platelets and gastric biopsies. The profound inhibition of monocyte COX2 activity at 24 h after dosing, as predicted by a pharmacological half-life of approximately 22 h, supports a once-daily dosing regimen of etoricoxib. In randomised, well-controlled clinical trials, etoricoxib has been shown to have a comparable clinical efficacy with traditional NSAIDs. Combined analysis of efficacy trials with etoricoxib versus non-selective NSAIDs has shown that the drug halves both investigator-reported upper gastrointestinal perforation, ulcers and bleeds (PUBs) and confirmed PUBs, and reduces the need for gastroprotective agents and gastrointestinal comedications by approximately 40%. The risk of lower extremity oedema and hypertension adverse experiences with etoricoxib was low and generally similar to comparator NSAIDs in a combined analysis of eight Phase III studies in OA, RA, chronic low-back pain and surveillance endoscopy. Large, randomised clinical trials have been planned to confirm the renal, gastrointestinal and cardiovascular safety of etoricoxib. PMID- 12562318 TI - Reviparin as prophylaxis for thromboembolism after leg injury and hip replacement. PMID- 12562319 TI - Is levosimendan a breakthrough in the development of positive inotropes? PMID- 12562320 TI - Phenotypic profile and functional characterization of rat lymph node-derived gammadelta T cells: implication in the immune response to cytomegalovirus. AB - Gammadelta T cells are unique, and their localization at sites of infection is considered critical in immune defence. We demonstrate the accumulation of gammadelta T cells in rat regional popliteal lymph nodes (PLNi) starting 2 days after inoculation of cytomegalovirus (CMV) into the footpad. Early-appearance PLNi gammadelta T cells significantly inhibited plaque development and the spread of CMV infection. These gammadelta T cells were negative for CD4 and CD8beta receptors, proliferated in response to interleukin-2 (IL-2) and contained high levels of interferon-gamma (IFN-gamma), the appearance of which correlated with the curing of fibroblasts from virus infection. The addition of anti-IFN-gamma abolished the ability of fibroblast monolayers to be cured from CMV infection. In contrast, this protection was not abolished by the addition of anti-rat IL-2 or anti-rat TNF-alpha, or by the depletion of NKR-P1-bearing cells within gammadelta T cells. In addition, the present study shows that while gammadelta T cells derived from naive and CMV-infected rats are able to kill both YAC-1 targets and CMV-infected syngeneic fibroblasts in vitro, only the latter are able to clear CMV-infected fibroblast monolayers. Finally, our data suggest that the expression of NKR-P1 by gammadelta T cells is critical for cytotoxicity, but its contribution to the curing from CMV infection was limited. PMID- 12562321 TI - Interleukin-18 plays a role in both the alum-induced T helper 2 response and the T helper 1 response induced by alum-adsorbed interleukin-12. AB - Previous studies have shown that the antigen-specific T helper 2 (Th2) response induced by alum adjuvants is interleukin (IL)-4 independent. As a role for IL-18 in Th2 induction has recently been described, in addition to its role in enhancing Th1 responses, we have studied the Th2 response induced by ovalbumin (OVA) adsorbed to alum in wild-type and IL-18-deficient mice. Our results indicate that while endogenous IL-18 facilitates alum-induced IL-4 production, OVA-specific immunoglobulin G1 (IgG1) and IgE production remain unaffected. Furthermore, antigen-specific Th1 responses induced with alum/IL-12-adsorbed OVA were demonstrated to be highly IL-18 dependent. Despite these observations, injection of BALB/c mice with exogenous IL-18 adsorbed to alum/OVA did not alter IL-4 or interferon-gamma production by T cells and had little effect on the relative production of IgG1/IgG2a antibody subclasses compared with alum/OVA inoculated mice. However, the previously described synergism between IL-12 and IL 18 in Th1 induction was evident as the Th1-promoting activity of alum/IL-12 against adsorbed OVA was greatly augmented by the coadministration of IL-18. These results indicate that while alum-induced IL-18 can facilitate Th2 induction, the addition of exogenous IL-18 cannot further enhance the alum induced Th2 response. PMID- 12562322 TI - Haptoglobin directly affects T cells and suppresses T helper cell type 2 cytokine release. AB - T helper cell type 1 (Th1) and type 2 (Th2) immune responses are characterized by a different pattern of cytokine expression following T-cell activation. Alterations of the ratio of Th1 to Th2 cells are important determinants of susceptibility to viral and parasitic infections, allergies, anti-tumour responses, and autoimmunity. In this work we bring new evidence for an effect of haptoglobin (Hp), a positive acute-phase protein, on T-lymphocyte functions. We show that Hp specifically interacts with both resting and activated CD4+ and CD8+ T cells. This specific binding results in a strong suppression of induced T-cell proliferation. In addition, Hp exhibits a strong in vitro inhibitory effect on Th2 cytokine release, while the production of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) is only slightly inhibited at high Hp doses. As a result, the presence of Hp promotes Th1 activation over Th2 activation in vivo as evidenced in Hp-deficient mice. Anti-CD3 monoclonal antibody injection indeed resulted in predominant IL-4 production in Hp-/- mice, in contrast to predominant IFN-gamma production in Hp+/+ mice. We conclude that Hp plays a modulating role on the Th1/Th2 balance by promoting a dominant Th1 cellular response. This points to a role of acute-phase proteins in balancing immune responses. PMID- 12562323 TI - The outcome of T-cell costimulation through intercellular adhesion molecule-1 differs from costimulation through leucocyte function-associated antigen-1. AB - Optimal T-cell activation requires both an antigen-specific and a costimulatory signal. The outcome of T-cell activation can be influenced by the nature of the costimulatory signal the T cell receives. We recently demonstrated the ability of stimulation through intercellular adhesion molecule-1 (ICAM-1), resident on the T cell surface, to provide a second signal for T-cell activation, and have extended that work here to begin an examination of the functional outcome of this set of signals. Costimulation through ICAM-1 resulted in a greater percentage of cells having undergone more than three divisions when compared to costimulation through leucocyte function-associated antigen-1 (LFA-1). Costimulation through ICAM-1 also had an effect similar to costimulation through CD28 in its ability to down regulate the cyclin dependent kinase inhibitor p27kip1. Costimulation through ICAM-1 provided greater protection from apoptosis than costimulation through LFA 1, especially in cells having divided more than three times. This was supported by the ability of costimulation through ICAM-1 to up-regulate the anti-apoptotic protein Bcl-2. Finally, costimulation through ICAM-1 or CD28 produced a greater number of T cells with a memory phenotype than costimulation through LFA-1. PMID- 12562324 TI - Roles for CD40, B7 and major histocompatibility complex in induction of enhanced immunity by cryptococcal polysaccharide-pulsed antigen-presenting cells. AB - Immunization of mice with activated antigen-presenting cells (APC) pulsed ex vivo with cryptococcal capsular polysaccharide, a glucuronoxylomannan (GXM-APC) results in prolongation of survival and delayed-type hypersensitivity (DTH) responsiveness following infection with Cryptococcus neoformans (NU-2). GXM-APC has both non-specific and GXM-specific effects that influence the immune responses that develop in mice after infection with NU-2. Type 1 cytokine responses are augmented after immunization with APC alone, while GXM must be present for the vaccine to influence survival and DTH reactions. This investigation evaluated the role that major histocompatibility complex (MHC) and co-stimulatory molecules play in the non-specific and GXM-specific responses induced by GXM-APC. APC from CD40 knockout mice were as effective as wild-type APC for the induction of non-specific and GXM-specific responses. Blocking activity of B7-1 and B7-2 by treatment of immunized mice with monoclonal antibodies specific for these molecules just before and for 6 days following GXM APC immunization decreased the splenic interferon-gamma response of mice subsequently infected with NU-2, but only in mice that were treated with both antibodies. These antibody treatments had no effect on DTH reactivity in similarly treated animals. MHC class I molecules were not involved in the antigen non-specific or GXM-specific activities of the vaccine. MHC class II molecules were not required for augmentation of type 1 cytokine responses but were needed for induction of the GXM-specific response that regulates the expression of DTH reactivity. This investigation has shown that an MHC class II-restricted, GXM specific response is responsible for altering DTH responsiveness which is the correlate of immunity in this model. PMID- 12562325 TI - Production of interleukin-13 by human dendritic cells after stimulation with protein allergens is a key factor for induction of T helper 2 cytokines and is associated with activation of signal transducer and activator of transcription-6. AB - Dendritic cells (DC) are able to induce not only T helper 1 (Th1) but also Th2 immune responses after stimulation with allergens. While DC-derived interleukin (IL)-12 and IL-18 are the key factors for the induction of Th1 cells, early signals being involved in Th2 differentiation are less well characterized so far. To analyse such early signals we used an antigen-specific setting with CD4+ T cells from atopic donors stimulated in the presence of autologous mature DC, which were pulsed with different allergen doses. The addition of increasing amounts of allergen during DC maturation with tumour necrosis factor-alpha, IL 1beta and prostaglandin E2 resulted in enhanced secretion of IL-6 and IL-12 by DC followed by increased production of Th1 (interferon-gamma; IFN-gamma) as well as Th2 (IL-4, IL-5) cytokines by CD4+ T cells. The coculture of allergen-treated DC and CD4+ T cells also led to a dose-dependent expression of active signal transducer and activator of transcription-6 (STAT6), which was visible already after 1 hr. Additionally, rapid phosphorylation of STAT6 was seen in immature DC after stimulation with allergens but not with lipopolysaccharide or human serum albumin. STAT6 phosphorylation was associated with the production of IL-13 by DC. The addition of neutralizing anti-IL-13 antibodies during maturation of DC inhibited STAT6 phosphorylation in CD4+ T cells as well as the production of IL 4, and to a lesser extent of IL-5, while IFN-gamma production was not affected. Addition of exogenous IL-13 enhanced mainly the secretion of IL-4. Taken together, DC-derived IL-13, which is released after exposure to allergens appears to be one of the critical factors for DC to acquire the capability to induce Th2 cytokine production. PMID- 12562326 TI - Tumour necrosis factor-alpha (TNF-alpha) transgene-expressing dendritic cells (DCs) undergo augmented cellular maturation and induce more robust T-cell activation and anti-tumour immunity than DCs generated in recombinant TNF-alpha. AB - Tumour antigen presentation by dendritic cells (DCs) to T cells in lymphoid organs is crucial for induction of anti-tumour immune responses. It has been previously reported that tumour necrosis factor-alpha (TNF-alpha) is required for DC activation and subsequent induction of optimal immune responses, and thus DCs for anti-tumour vaccination are often generated by culture in exogenous TNF alpha. In the present study, we investigated the effect on anti-tumour immunity of vaccination with Mut1 tumour peptide-pulsed DCs engineered to express a TNF alpha transgene. Our data shows that transfection of DCs with recombinant adenovirus AdV-TNF-alpha resulted in greater maturation of the DCs than occurred with control DCs cultured in exogenous TNF-alpha, as determined by up-regulated expression of pro-inflammatory cytokines (e.g. interleukins 1beta and 18), chemokines [e.g. interferon-gamma-inducible protein-10 and macrophage inflammatory protein-1beta (MIP-1beta)], the CC chemokine receptor CCR7, and immunologically important cell surface molecules (CD40, CD86 and intercellular adhesion molecule-1). These transgenic DCs stimulated stronger allogeneic T-cell responses in vitro and T-cell activation in vivo; displayed 2.4-fold enhanced chemotactic responses to the MIP-3betain vitro (P<0.05); and, perhaps most importantly, trafficked into the draining lymph nodes dramatically (seven-fold, P<0.01) more efficiently than the control DCs. Our data also demonstrate that vaccination of mice with Mut1 peptide-pulsed, AdV-TNF-alpha-transfected DCs stimulated more efficient in vitro Mut1-specific CD8+ cytotoxic T-cell responses and solid tumour immunity in vivo, when compared to the in vitro TNF-alpha cultivated DCs. Thus, DCs engineered to secrete TNF-alpha may offer a new strategy in DC cancer vaccines. PMID- 12562327 TI - Breast milk macrophages spontaneously produce granulocyte-macrophage colony stimulating factor and differentiate into dendritic cells in the presence of exogenous interleukin-4 alone. AB - Peripheral blood monocytes extravasate and differentiate into tissue macrophages to mediate effective local defence, but how tissue-specific stimuli and environments may influence their functions remains unknown. Here, we found that peripheral blood monocytes gained the ability to produce granulocyte-macrophage colony-stimulating factor (GM-CSF) upon exposure to breast milk and differentiated into CD1+ dendritic cells (DCs) in the presence of exogenous interleukin-4 (IL-4) alone. This in vitro observation appeared physiologically relevant since macrophages that were freshly isolated from breast milk were also found to produce GM-CSF spontaneously. Furthermore, in contrast to peripheral blood monocytes that differentiated into DCs only in the presence of both exogenous GM-CSF and IL-4, differentiation of breast milk macrophages into DCs was induced by incubation with exogenous IL-4 alone. These IL-4-stimulated breast milk macrophages were efficient in stimulating T cells, suggesting their potential role in mediating T-cell-dependent immune responses in situ. On the other hand, unexpected expression of DC-SIGN, a DC-specific receptor for human immunodeficiency virus (HIV), even in unstimulated breast milk macrophages, may favour HIV infection, resulting in an increased risk of mother-to-infant vertical transmission of the virus via breast milk. Thus, tissue-specific development of macrophages is often linked to effective local immunity, but may potentially provide an opportunity for a pathogen to spread and transmit. PMID- 12562328 TI - Administration of exogenous interleukin-18 and interleukin-12 prevents the induction of oral tolerance. AB - Interleukin-18 (IL-18), a pro-inflammatory member of the IL-1 family, has been associated with autoimmunity and allergic disease. This type of pathology is thought to be the result of a defect in immunological tolerance and is often observed in inflammatory disorders of the gut such as coeliac disease, Crohn's disease and ulcerative colitis. IL-18 has been implicated in a number of mucosal immune disorders, where it synergizes with IL-12 to induce the production of interferon-gamma (IFN-gamma). We have therefore investigated the effects of IL-18 and IL-12 administration on the induction of oral tolerance to ovalbumin. The suppression of specific Ig G2a production, delayed-type hypersensitivity responses and IFN-gamma production by antigen-specific T cells were all abrogated by the presence of exogenous IL-12 and IL-18, suggesting that oral tolerance was broken. The expression of the co-stimulatory molecule CD80 on dendritic cells was also shown to be increased by this combination of cytokines. As dendritic cells are thought to be of major importance in the induction of tolerance, this suggests a mechanism by which tolerance to mucosal antigens may be broken in vivo. PMID- 12562329 TI - Regulation of epithelial cell cytokine responses by the alpha3beta1 integrin. AB - Epithelial cells (EC) from various tissues can produce important cytokines and chemokines when stimulated by proinflammatory cytokines. These EC also receive signals from cell surface integrins, like the alpha3beta1 integrin, which is important in cell migration and wound healing of epithelial monolayers. However, little is known of the effect of integrin signals on cytokine responses by EC. Colonic Caco-2 cells treated with an anti-alpha3 integrin antibody prior to stimulation with the proinflammatory cytokine interleukin (IL)-1 yielded suppressed levels of mRNA and secreted IL-6, IL-8 and monocyte chemoattractant protein-1 as compared to cells treated with normal mouse immunoglobulin G. Lung A549 cells also showed a similar suppression of cytokine secretion. Likewise, treatment of the Caco-2 cells with the same antibody suppressed tumour necrosis factor-alpha-stimulated IL-6 secretion. Fab fragments of the anti-alpha3 integrin antibody did not induce the suppressive effect but did block the suppressive effect of the whole antibody suggesting that the effect of the antibody required cross-linking of the integrins. Finally, culture of the Caco-2 cells on laminin type 5 (the major ligand for this integrin) yielded depressed levels of IL-1 induced IL-6 secretion as compared to cells on laminin type 1. These data are the first indication that the alpha3beta1 integrin may cause a suppression of cytokine responses by EC which may be important in regulating the capacity of EC to respond during inflammation or wound healing. PMID- 12562330 TI - Functional and Vbeta repertoire characterization of human CD8+ T-cell subsets with natural killer cell markers, CD56+ CD57- T cells, CD56+ CD57+ T cells and CD56- CD57+ T cells. AB - We investigated the individual CD8+ populations with natural killer (NK) cell markers (NK-type T cell); CD56 single positive (CD56)-T cells, CD56/CD57 double positive (DP)-T cells and CD57 single positive (CD57)-T cells in the peripheral blood. All NK-type T-cell populations expressed CD122 and intermediate levels of T-cell receptor (TCR; regular CD8+ T cells are CD122- and express high levels of TCR). The number of both DP-T cells and CD57-T cells, but not CD56-T cells, gradually increased with age. All NK-type T-cell populations produced larger amounts of interferon-gamma than did regular CD8+ T cells after stimulation with interleukin (IL)-2, IL-12 and IL-15. However, CD56-T cells and CD57-T cells but not DP-T cells showed a potent antitumour cytotoxity to NK-sensitive K562 cells, whereas only CD56-T cells showed a potent cytotoxity to NK-resistant Raji cells. Furthermore, although NK-type T cells produced large amounts of soluble Fas ligands, their cytotoxic activities appeared to be mediated by the perforin/granzyme pathway. The oligoclonal or pauciclonal expansions of certain VbetaT cells were found in each NK-type T-cell population. The non-variant CDR3 region(s) for the TCRbeta chain(s) showed CD57-T cells and CD56-T cells to be derived from distinct origins, while the DP-T cell population consisted of a mixture of the clones seen in both CD56-T cells and CD57-T cells. Our results suggest that CD57-T cells and CD56-T cells are functionally and ontogenically different populations while DP-T cells appear to originate from both CD56-T cells and CD57-T cells. PMID- 12562331 TI - Recognition of bovine respiratory syncytial virus proteins by bovine CD8+ T lymphocytes. AB - CD8+ T lymphocytes play a major role in the clearance of bovine respiratory syncytial virus (BRSV), an important respiratory pathogen of young calves that shares many of the epidemiological and pathological features of human respiratory syncytial virus (HRSV) in infants. Recombinant vaccinia virus (rVV) and recombinant fowlpox virus (rFPV), expressing individual BRSV proteins, were used to demonstrate that the F, N and M2 proteins were the major antigens recognized by bovine CD8+ T cells in major histocompatibility complex (MHC)-defined cattle. BRSV protein recognition by CD8+ T cells was analysed using cytotoxic T lymphocyte (CTL) assays or by the production of interferon-gamma (IFN-gamma) following restimulation with BRSV proteins. Strong recognition of the G protein by CD8+ T cells was observed in cattle that had been vaccinated with rVV expressing this protein and subsequently challenged with BRSV. Although there is variation in the number of expressed MHC genes in cattle with different class I haplotypes, this did not appear to influence BRSV protein recognition by CD8+ T cells. Knowledge of the antigenic specificity of BRSV-specific CD8+ T cells will facilitate the qualitative and quantitative analysis of BRSV-specific CD8+ T-cell memory in cattle and help to ensure that potential vaccines induce a qualitatively appropriate CD8+ T-cell response. PMID- 12562332 TI - Resistant mice lacking interleukin-12 become susceptible to Trypanosoma cruzi infection but fail to mount a T helper type 2 response. AB - Interleukin-12 (IL-12) is essential to resistance to Trypanosoma cruzi infection because it stimulates the synthesis of interferon-gamma (IFN-gamma) that activates macrophages to a parasiticidal effect. Investigation of mice deprived of IL-12 genes (IL-12 knockout mice) has confirmed the important role of IL-12 and IFN-gamma in controlling parasitism in T. cruzi infection. However, it has not yet been addressed whether a shift towards a T helper type 2 (Th2) pattern of cytokine response occurred in these mice that might have contributed to the aggravation of the infection caused by IL-12 deprivation. We examined the course of T. cruzi (Y strain) infection and the regulation of cytokine responses and nitric oxide production in C57BL/6 IL-12 p40-knockout mice. The mutant mice were extremely susceptible to the infection as evidenced by increased parasitaemia, tissue parasitism and mortality in comparison with the control C57BL/6 mouse strain (wild-type) that is resistant to T. cruzi. A severe depletion of parasite antigen-specific IFN-gamma response, without an increase in IL-4 or IL-10 production, accompanied by reduced levels of nitric oxide production was observed in IL-12 knockout mice. We found no evidence of a shift towards a Th2-type cytokine response. In IL-12 knockout mice, the residual IFN-gamma production is down-regulated by IL-10 but not by IL-4 and nitric oxide production is stimulated by tumour necrosis factor-alpha. Parasite-specific immunoglobulin G1 antibody levels were similar in IL-12 knockout and wild-type mice, whereas IL-12 knockout mice had much higher levels of immunoglobulin G2b. PMID- 12562338 TI - Effect of wax epilation before hair removal with a long-pulsed alexandrite laser: a pilot study. AB - BACKGROUND: Recent reports indicate that laser hair removal is most effective on anagen hairs. However, no published trials have examined laser epilation after hair cycle synchronization. OBJECTIVE: To evaluate the potential for enhanced laser hair removal after the induction of telogen hairs into anagen by wax epilation. METHODS: We identified four 2.5-cm square areas with equivalent hair length and density on the backs of 13 dark-haired white men. To induce typically telogen hairs into anagen, two areas on each patient were wax epilated. Two weeks later, one waxed area and one unwaxed area were treated with a long-pulsed alexandrite laser. One month after laser treatment, a subjective comparison was made based on hair density, length, and thickness. RESULTS: In 12 of 13 patients, lasered areas that had been pretreated with wax epilation were clearer of hair as compared with areas that had been pretreated by shaving (P=0.0034). No significant difference was noted between waxed and unwaxed control areas that had not been laser treated (P=1.0). CONCLUSION: Wax epilation 2 weeks before laser hair removal improves cosmetic outcomes at 1 month. This effect may be secondary to the recruitment and heightened sensitivity of early anagen hairs. PMID- 12562333 TI - Inducible nitric oxide synthase deficiency in mice increases resistance to chronic infection with Echinococcus multilocularis. AB - The production of nitric oxide (NO) by intraperitoneal macrophages of mice during secondary infection with Echinococcus multilocularis mediates immunosuppression at early and late stages of infection. We addressed the role of NO in host resistance against this extracellular metazoan parasite by infecting inducible nitric oxide synthase knockout ((iNOS-KO) mice (of the C57BL/6 background) with 100 metacestode vesicles. The parasite weight was significantly lower in iNOS-KO mice when compared with wild-type (WT) mice at 4 months postinfection (late stage), thus demonstrating that iNOS deficiency confers a certain degree of resistance against persistent chronic infection. However, histological analysis of periparasitic tissue showed no differences between WT and iNOS-KO mice, as both exhibited granuloma formation and the presence of giant cells. Together with histology, the production of a high level of interferon-gamma (IFN-gamma) in infected iNOS-KO mice upon stimulation with concanavalin A (Con A) and VF-antigen indicated normal T-cell signalling in these animals. As expected, peritoneal exudate cells (PEC) from infected iNOS-KO mice produced no detectable NO, while the PEC from infected WT mice produced high levels of NO after stimulation with lipopolysaccharide (LPS) and parasite protein or carbohydrate antigen, or even without in vitro stimulation. Consequently, the high level of NO production observed during chronic infection in WT mice appears to contribute more to immunosuppression than to limitation of parasite growth. This is also reflected by the fact that splenocyte proliferation was significantly higher and parasite masses lower in iNOS-KO mice (at 1 and 4 months postinfection) than in WT mice. PMID- 12562334 TI - A parallel signal-transduction pathway for eotaxin- and interleukin-5-induced eosinophil shape change. AB - Interleukin-5 (IL-5) and eotaxin are the most important cytokines/chemokines responsible for regulating eosinophil locomotion and are known to play a co operative role in the selective recruitment of eosinophils to inflamed tissues. Following exposure to chemoattractants, eosinophils undergo a series of events, including reorganization of actin filaments and subsequent rapid shape changes, culminating in chemotaxis. In this study we examined the signalling pathways for eosinophil shape change regulated by eotaxin and IL-5, primarily using a gated autofluorescence/forward-scatter assay. Eotaxin and IL-5 were able to elicit shape change with peaks at 10 and 60 min, respectively, and IL-5 triggered the shape change more efficiently than eotaxin. The pharmacological inhibitors of mitogen-activated protein kinase (MAP kinase) and p38 blocked both eotaxin- and IL-5-induced eosinophil shape change in a dose-dependent manner. In addition, depletion of intracellular Ca2+ and inhibition of protein kinase A (PKA) strongly reduced eosinophil shape change. In contrast, even when used at high concentrations, protein tyrosine kinase (PTK) inhibitors caused only a slight reduction in the ability to change shape. However, treatment with protein kinase C (PKC) inhibitors, such as GF109203X and staurosporine, resulted in a striking inhibition of eosinophil shape change by IL-5, but not eotaxin. Data from the inhibition of activation and chemotaxis of the extracellular signal-regulated kinases (ERK1/2) by the PKC inhibitors were also consistent with findings from the experiments on shape change. Collectively, two eosinophil-selective cytokines/chemokines probably regulate eosinophil shape change via a largely overlapping signalling pathway, with involvement of PKC restricted to the IL-5 signal alone. PMID- 12562339 TI - Treatment of facial rhytides with a nonablative 1,450-nm diode laser: a controlled clinical and histologic study. AB - OBJECTIVE: To evaluate the safety and effectiveness of a nonablative 1,450-nm midinfrared diode laser in the treatment of facial rhytides. METHODS: Twenty-five patients (skin phototypes I-III) with mild-to-moderate perioral or periorbital rhytides received four successive treatments at 3- to 4-week intervals with a 1,450-nm diode laser (SmoothBeam; Candela Corp., Wayland, MA). Symmetrical matched areas were left untreated to serve as controls. Patients were evaluated with digital photography at each treatment session and at 1, 3, 6, and 12 months after the final laser treatment. Skin biopsies were obtained from treatment and control facial sites for histologic evaluation before treatment, immediately after the first treatment, and 3, 6, and 12 months after the fourth treatment. Patient satisfaction scores were obtained at each follow-up visit. RESULTS: Mild to moderate improvement in treated facial rhytides was observed in all patients. Increased dermal collagen was seen 6 months after four successive treatments. Patient satisfaction scores paralleled the photographic and histopathologic changes seen. Side effects were limited to transient erythema, edema, and postinflammatory hyperpigmentation. CONCLUSION: The nonablative 1,450-nm diode laser is safe and effective for the treatment of mild-to-moderate facial rhytides. PMID- 12562340 TI - Lymphocytic infiltrates and subclinical epithelial tumor extension in patients with chronic leukemia and solid-organ transplantation. AB - BACKGROUND: Dense infiltrates in association with squamous cell carcinoma (SCC) or basal cell carcinoma (BCC) in patients with underlying chronic lymphocytic leukemia (CLL) may complicate pathologic interpretation of histologic margins. OBJECTIVE: The study was conducted to determine the frequency of identifying dense inflammatory infiltrates in frozen histologic sections during Mohs operation for BCC or SCC in patients with CLL and organ-transplant recipients, to characterize the infiltrate (reactive versus leukemic) in CLL, and to estimate the subclinical tumor extension in patients with CLL, transplant recipients, and control subjects undergoing Mohs procedure. METHODS: Frozen sections of head and neck BCC and SCC obtained during Mohs procedures in patients with CLL, organ transplant recipients, and a control group were reviewed retrospectively. Biopsy specimens of CLL with dense infiltrates were assessed with immunohistochemical stains. Subclinical tumor extension (postoperative defect size minus preoperative tumor size) was evaluated in each group. RESULTS: Dense infiltrates were found in tumors of 20 of 55 patients with CLL (36%), 1 of 8 transplant recipients (13%), and 1 of 105 controls (1%). In patients with CLL, 75% of the dense infiltrates were B-cell leukemic. Compared with controls, the mean subclinical tumor extension was larger in patients with CLL (P=0.029) and in transplant recipients (P=0.55). CONCLUSION: Dense leukemic infiltrates associated with BCC or SCC in CLL may complicate pathologic interpretation of Mohs surgical histologic margins and may be associated with larger postoperative defects relative to preoperative clinical tumor appearance. In patients with CLL, as in transplant recipients, SCC seems more likely to develop than BCC. PMID- 12562341 TI - Utility of sentinel lymphadenectomy in the management of patients with high-risk cutaneous squamous cell carcinoma. AB - BACKGROUND: High-risk cutaneous squamous cell carcinoma (SCC) is an aggressive tumor with a significant rate of metastasis, especially nodal, and deaths yearly. Sentinel lymphadenectomy may be a valuable adjunct in the treatment of patients with cutaneous SCC at high risk for metastases with a clinical N0 status. OBJECTIVE: To report the clinical and pathologic features in nine patients who underwent this procedure at a tertiary-care cancer center. METHODS: Since 1995, a total of nine patients from the Cutaneous Oncology Program at the H. Lee Moffitt Cancer Center received preoperative lymphoscintigraphy and sentinel lymphadenectomy for high-risk cutaneous SCC with a clinical N0 status. RESULTS: Histologically positive nodes were found in 4 of 9 cases (44%). Two of the four patients with positive sentinel nodes died of metastatic disease within 2 years. All five patients with negative sentinel nodes are alive and well at a median follow-up of 8 months (mean of 13 months). Preoperative lymphoscintigraphy and sentinel lymphadenectomy were well tolerated by all patients. CONCLUSION: In this small series of predominantly trunk and extremity high-risk SCCs, sentinel lymph node biopsy was technically feasible with low morbidity. Sentinel lymphadenectomy may prove to have an important role in the management of high-risk cutaneous SCC with a clinical N0 status. PMID- 12562342 TI - Sentinel node detection in malignant melanoma patients: radiation safety considerations. AB - BACKGROUND: The surgical management of malignant melanoma necessitates correct sentinel lymph node localization. The highest reported sensitivities are those of lymphoscintigraphy and intraoperative gamma-probe detection combined with a vital blue dye technique. OBJECTIVE: Control of the radiation doses experienced by surgical personnel untrained in the use of unsealed radioactive materials. METHODS: Sentinel lymph nodes were localized, and biopsies were performed in 25 patients with malignant melanoma. Radiation doses during surgery were determined with energy-compensated silicon pin diode detectors and LiF thermoluminescent ring dosimeters. RESULTS: In 21 cases (24%), the measured doses were less than 1 microSv, but in 4 operations (16%), 1 to 4.5 microSv was received. The equivalent dose rate was generally less than 1 microSv/h. The finger-absorbed doses for the surgeon and the assistant surgeon were (mean+/-SD) 159+/-23 and 48+/-17 microGy per intervention, respectively. CONCLUSION: Personal dosimetric survey and limitation of the number of surgical interventions do not appear to be essential. PMID- 12562343 TI - Importance of subfascial collaterals in deep-vein malformations. AB - BACKGROUND: The removal of varicose veins in patients with absent or hypoplastic deep veins is contraindicated. OBJECTIVE: To determine a reliable examination for detection of the function of subfascial collaterals. METHODS: Five crural ulcer limbs with deep-vein aplasia and hypoplasia were operated on. In all cases, a segmental or total deep-vein aplasia (four cases) or hypoplasia (one case) was found in the affected limb by venography. Patients were selected for surgery using a new modification of the Perthes test. Radical varicectomy was performed in every case using stripper and open varicectomy. RESULTS: There was no circulatory complication during or after the operation. Crural ulcers healed. CONCLUSION: The agenesia or hypoplasia of a shorter or longer segment of deep veins is not a contraindication to radical varicectomy in every patient. PMID- 12562344 TI - Coding multiple diagnoses for patient visits at which procedures were performed: no evidence for abuse by physicians. AB - BACKGROUND: A problem area in the reimbursement of physicians is the office visit at which both procedures and evaluation/management services are performed. Insurers are concerned that frequent reporting of diagnoses unrelated to procedures may represent an abuse of the claims process. OBJECTIVE: To determine the frequency of secondary diagnoses that are present at visits at which procedures are performed and to compare the frequency with that reported in Medicare claims. METHOD: The 1998 and 1999 National Ambulatory Medical Care Survey (NAMCS) data were used to provide data unrelated to claims for payment. The results were compared with Medicare claims data from the 1998 to 1999 Medicare Current Beneficiary Survey. NAMCS visits were limited to patients who were 65 year old or greater to limit the analysis to the Medicare population. RESULTS: In the NAMCS dataset, 52% of the visits for actinic keratoses at which a procedure was performed were associated with multiple diagnoses. Similarly, in the Medicare Current Beneficiary Survey dataset, 52% of these visits had multiple diagnoses listed. CONCLUSION: The frequency of visits with multiple diagnoses is the same in both administrative claims and epidemiologic survey databases. There is no evidence for widespread abusive coding by physicians. It is incumbent on payers to respect the integrity of the coding system in order to assure fair reimbursement for physician services. PMID- 12562345 TI - Clinical trial of bleaching treatment with 10% all-trans retinol gel. AB - BACKGROUND: Although an aggressive use of tretinoin along with hydroquinone enables an efficient treatment of hyperpigmented skin lesions, irritant dermatitis remains to be solved. OBJECTIVE: To evaluate the efficiency and adverse effects of 10% all-trans retinol (ROL) gel for improvement of skin hyperpigmentation. METHODS: Ten-percent ROL gel was used instead of 0.1% tretinoin gel in our two-phased bleaching protocol (bleaching and healing phases); 5% hydroquinone and 7% lactic acid ointment were used along with ROL gel in the bleaching phase (2 to 6 weeks). Five-percent hydroquinone and 7% ascorbic acid ointment were used alone during the healing phase (4 to 6 weeks). Twenty-one Japanese patients with hyperpigmented lesions on the face were enrolled in this study, and 18 patients who were followed for more than 10 weeks were analyzed. RESULTS: Improvement of pigmentation was seen in 16 of 18 patients after an average treatment period of 11.3 weeks, and in 6 patients, pigmentation was almost eliminated after treatment. Erythema and scaling were seen, however, during the bleaching phase as well as the bleaching treatment with tretinoin gel. CONCLUSION: ROL can improve skin hyperpigmentation to a similar extent to tretinoin when used at high concentration, whereas it induces irritant dermatitis as well. PMID- 12562346 TI - Nonablative dermal remodeling with a 585-nm, 350-microsec, flashlamp pulsed dye laser: clinical and ultrastructural analysis. AB - BACKGROUND: A nonablative 350-microsec, 585-nm, flashlamp pulsed dye laser is currently being used for the treatment of rhytides. OBJECTIVE: To analyze both clinical rhytid improvement and electron microscopic evidence of ultrastructural changes after treatment with a nonablative 350-microsec, 585-nm, flashlamp pulsed dye laser. RESULTS: At 6 months after two treatments, 40% of the treated subjects noted mild improvement in rhytid appearance. Nontreating physician evaluation revealed some degree of improvement in 50% of the treated subjects. Mild improvement in quality and texture of the skin was also reported by 50% of the subjects. Electron microscopic evaluation showed ultrastructural changes that are consistent with new collagen formation. CONCLUSION: Treatment with a nonablative 350-microsec, 585-nm, flashlamp pulsed dye laser can lead to both clinical and electron microscopic evidence of improvement in photo-damaged skin. PMID- 12562347 TI - Breast enlargement observed after power liposuction: a retrospective review. AB - BACKGROUND: After undergoing power liposuction to various body areas, a significant number of subjects reported an increase in their breast size that occurred spontaneously several months after their procedure. OBJECTIVE: To establish the incidence of breast enlargement among subjects who underwent power liposuction and to identify variables that are associated with the phenomenon. METHODS: A retrospective chart review and patient interview were performed from among 73 subjects who had undergone power liposuction at our center. Variables such as age, original weight, volume of aspirated fat, estrogen supplements, amount of weight loss or gain, and body areas aspirated were compared between those who experienced breast enlargement and those who did not. RESULTS: Thirty four percent of the subjects (25 of 73) reported an increase in breast size. Of this group, 32% (8 of 25) reported an increase in cup size of one or more. Variables associated with the cohort who experienced breast enlargement included larger volume of fat aspiration and liposuction in the abdomen and hip location. CONCLUSION: Breast enlargement was observed in 34% of the subjects after power liposuction either in the form of a larger bra cup size or a subjective feeling of increased fullness. We hypothesize that an altered androgen to estrogen ratio after liposuction may be responsible for this change. PMID- 12562348 TI - Use of honey as an adjunct in the healing of split-thickness skin graft donor site. AB - BACKGROUND: Different techniques are being used in treatment of split-thickness skin graft donor sites; however, there is not a widely accepted method established for these partial-thickness wounds. It is well known that honey has been very effective in the treatment of various types of wounds, but there is not any information about the usage of honey as split-thickness skin graft donor site dressing in the literature. OBJECTIVE: To evaluate and compare the effectiveness of honey-impregnated gauzes, hydrocolloid dressings, and as a conventional dressing, saline-soaked gauzes for skin graft donor sites. METHODS: This is a nonrandomized, prospective, open-label (noncontrolled), side-by-side comparison trial of various options that are available for second-intention healing of donor site for split-thickness skin grafts. Eighty-eight patients who underwent skin grafting were observed using two different groups. In the first group, the donor site was divided into two equal halves, with each half being treated with honey soaked gauzes and the other half with paraffin gauzes (group 1A), hydrocolloid dressings (group 1B), and saline-soaked gauzes (group 1C) alternatively. In the second group, two separate donor sites were formed, with one of them being treated with honey-impregnated gauzes (groups 2A-C) and the other one treated with either paraffin gauzes (group 2A), hydrocolloid dressings (group 2B), or saline-soaked gauzes (group 2C). The healing time, rate of infection, and sense of pain were evaluated. RESULTS. : In the treatment of split-thickness skin graft donor sites, honey-impregnated gauzes showed faster epithelization time and a low sense of pain than paraffin gauzes and saline-soaked gauzes. There was no significant difference between honey-impregnated gauzes and hydrocolloid dressings with regard to epithelization time and sense of pain. CONCLUSION: The use of honey-impregnated gauzes is effective, safe, and practical. Honey can be an alternative material for the split-thickness skin graft donor site treatment. PMID- 12562349 TI - Surgical treatment of chronic gluteal hidradenitis suppurativa: reused skin graft technique. AB - BACKGROUND: The treatment of chronic lesions in hidradenitis suppurativa (HS) remains a challenge for dermatologists. In most cases, wide surgical excision of the affected skin reduces the recurrence rate to a minimum. Split-thickness skin grafts have usually been applied to resurface large postoperative defects. OBJECTIVE. : The aim of this study is to introduce an alternative method of skin grafting, called "reused" or "recycled" skin graft, for the reconstruction of the large skin defect with chronic gluteal HS. METHODS: The study consisted of six patients (two females and four males) with gluteal HS. After a wide en bloc excision, the wound was immediately recovered with meshed-skin graft, made from the resected skin itself. Thus, the sacrifice of the skin donor is spared. The drum dermatome (Padgett-Hood) is suitable to take the split-skin graft from the resected skin of the affected buttock. The thickness of grafts was set between 12/1,000 and 20/1,000 inches, and all grafts were meshed with 1.5 times the expansion. The skin grafts were secured in place on the wound and a tie-over dressing was applied. RESULTS: Postoperative complications were usually minor ones, such as hematoma, discharge, and small areas of graft skin necrosis (less than 1 cm2), although one patient developed a 3 x 4 cm2 graft necrosis and wound infection. The follow-up period after surgery ranged from 8 to 36 months. No patient experienced any functional disabilities or recurrence during follow-up years. CONCLUSION: When the epidermal involvement remains mild to moderate, this reused skin graft technique is an alternative choice to resurface the surgical defect of gluteal HS. It is superior to the conventional procedure, which requires fresh skin donor site. PMID- 12562350 TI - Chondroid syringoma: a diagnosis more frequent than expected. AB - BACKGROUND: Chondroid syringoma or mixed tumor of the skin is a rare subcutaneous tumor that may be confused with various skin lesions. OBJECTIVE: To elucidate the incidence of condroid syringomas among skin lesions that were excised under local anesthesia. METHODS: The histopathologic diagnosis of 16,200 skin lesions that had been operated between 1986 and 2002 were retrospectively evaluated. The cases with condroid syringoma were histopathologically re-examined, and confirmed cases were further analyzed for preoperative diagnosis, age, gender, and lesion location. RESULTS: Sixteen patients were found to have histopathologic diagnosis of chondroid syringoma constituting 0.098% of the excised skin lesions in this series. All of these 16 cases were misdiagnosed preoperatively. The typical presentation was a solitary skin lesion located in the head and neck region in a middle-aged male patient. CONCLUSION: In the evaluation of a middle-aged male patient with a small subcutaneous nodule in the head and neck region, chondroid syringoma should be also taken into consideration for differential diagnosis. For such a lesion, excisional biopsy without destroying aesthetic and functional structures is the preferred diagnostic approach. PMID- 12562351 TI - Repair of a large surgical defect involving the lower lid, cheek, and temporal region. PMID- 12562352 TI - Treatment of toxic epidermal necrolysis with moisture-retentive ointment: a case report and review of the literature. AB - Toxic epidermal necrolysis (TEN) is a rare condition that was described by Lyell in 1956. It is a severe, acute, adverse, primarily drug-induced, potentially fatal, cutaneous reaction that is characterized by large areas of skin desquamation and sloughing, similar in many aspects to second-degree burns. The treatment of cutaneous drug reactions rests essentially on immediate diagnosis and recognition of the disease process, accurate history, thorough physical examination, prompt discontinuation of the offending drug, and supportive care. TEN patients are best managed in specialized burn units. Nevertheless, the management remains very much individualized, based on the clinical setting. Topical wound care remains an essential factor in the treatment of burn-like syndromes and is a main determining parameter for morbidity and mortality. As the value of moist environment in wound healing is being fully appreciated, we report on the use of a newly introduced ointment, the Moist Exposed Burn Ointment (Julphar; Gulf Pharmaceutical industries, Ras El-Khaymah, United Arab of Emirutes), a moisture-retentive ointment, in the successful management of a case of TEN. PMID- 12562353 TI - Successful treatment of a hypochromic scar with manual dermabrasion: case report. AB - BACKGROUND: Manual dermabrasion is a simple and safe cosmetic procedure that is used to improve unsightly cutaneous disorders. OBJECTIVE: To describe an 11-year old white female patient who was successfully treated with manual dermabrasion for a hypochromic scar on the left forearm. RESULTS: The hypochromic wound healed without scar and with good cosmetic appearance. CONCLUSION: Although similar to conventional dermabrasion in terms of cosmetic results, manual dermabrasion presents a less expensive and safe alternative. PMID- 12562354 TI - On glomus tumors, warts, and razors. AB - BACKGROUND: Occam's razor encourages ascribing a set of clinical findings to a single diagnosis. OBJECTIVE: To present a patient with a subungual wart and a glomus tumor of the same nail unit and to review the literature regarding these conditions. METHODS: We describe a 58-year-old female with a 2-year history of nail dystrophy and related symptoms that were initially attributed to verruca but recalcitrant to extensive therapies. RESULTS: Ultimately the persistence of her symptoms was found to be secondary to a subungual glomus tumor. CONCLUSION: Clinicians must maintain a high index of suspicion for the presence of multiple diagnoses to prevent the delay of appropriate management, particularly when symptoms do not respond to initial treatment. PMID- 12562355 TI - Unusual herpes simplex virus infection mimicking foreign body reaction after cosmetic lip augmentation with expanded polytetrafluoroethylene threads. AB - BACKGROUND: Adverse reactions after cosmetic lip augmentation with synthetic implant materials include inflammation, infection, and extrusion of and foreign body reaction to the implant. Clinically, all of them are characterized by pain, erythema, swelling, and formation of papules and/or pustules. OBJECTIVE: To describe an unusual herpes simplex virus infection in a 48-year-old female patient after cosmetic lip augmentation with expanded polytetrafluoroethylene threads clinically resembling a foreign body reaction. METHODS: Case report and literature review. RESULTS: A histopathologic examination showed no signs of a foreign body reaction. Instead, herpes simplex virus-specific DNA was demonstrated by polymerase chain reaction from lesional tissue. CONCLUSION: Cosmetic lip augmentation with expanded polytetrafluoroethylene threads may be complicated by a delayed local herpes simplex virus infection clinically mimicking a foreign body reaction to the implant. PMID- 12562356 TI - Cutaneous metaplastic synovial cyst: unusual presentation with "a bag of worms". AB - BACKGROUND: Cutaneous metaplastic synovial cyst (CMSC) is a rare cystic tumor that is unfamiliar to most dermatologists. A traumatic history usually precedes its onset. CMSC is often misdiagnosed as suture granuloma clinically. A correct diagnosis requires histopathologic examination to reveal its characteristic features, that is, a cystic tumor line by synovium-like structure. OBJECTIVE: To describe a case of CMSC encountered during dermatologic surgery practice. We present an unusual case with elongated projections. METHODS: A case report and literature review are presented. CONCLUSION: The differential diagnosis of tender cutaneous cysts should include CMSC. Projections from the cyst wall may attain significant size and give the intraoperative impression of a "bag of worms." PMID- 12562357 TI - Rapid development of keratoacanthomas after a body peel. AB - Resurfacing techniques have been traditionally limited to the face because of a lack of predictability and standardization for peeling nonfacial skin. There is a need for medical and surgical intervention for treating nonfacial skin that is actinically damaged. Medium-depth chemical peels (Jessner +35% trichloroacetic acid) remove the photodamaged epidermis to stimulate the production of new collagen in the dermis and remove lesions associated with facial actinic damage, including lentigines and actinic keratoses. Widespread actinic damage is common on the arms and chest. A 70% glycolic acid gel plus 40% trichloroacetic acid peel (Cook Body Peel) is a controlled peel that predictably enables peeling of nonfacial skin in a uniform and safe fashion with specific clinical endpoints. An unusual complication of this body peel is reported. PMID- 12562358 TI - Melasma in Indian males. PMID- 12562359 TI - Appositional bone formation in marginal defects at implants. AB - In a previous experiment, it was demonstrated that a wide marginal defect around an implant can heal with a high degree of osseointegration. The present experiment was performed to evaluate the degree and quality of de novo bone formation and osseointegration in marginal defects adjacent to submerged titanium implants. All mandibular premolars and 1st molars were extracted in four Labrador dogs. Four experimental sites were identified in the right side of the mandible. In two sites, custom-made implants with a sandblasted, large grit, acid-etched (SLA) surface were installed without further ostectomy (control sites). In the two remaining sites (test sites), a specially designed step drill was used to widen the marginal 5 mm of the canal. A barrier membrane was used to cover the implants in the defect sites. All implants were submerged. One month later, an identical procedure, including site preparation and implant installation, was performed in the left side of the mandible. Two months following the first implant installation procedure, biopsies were collected and prepared for sectioning. Ostectomy and implant installation in the control location resulted in a series of bone tissue alterations which eventually allowed newly formed bone to establish contact with the SLA surface. The marginal defect lateral to the implant in the test locations gradually became filled with newly formed bone. De novo bone formation started within the walls of the surgically prepared defect. Bone-to-implant contact was first established in the apical portion of the gap. This new bone tissue was in the coronal direction continuous with a dense, non mineralized 'implant attached' soft tissue which, over time, also became mineralized to increase the height of the zone of bone-to-implant contact. The results suggest that healing of a wide marginal defect around an implant is characterized by appositional bone growth from the lateral and apical bone walls of the defect. PMID- 12562360 TI - IL-1 gene polymorphism and smoking as risk factors for peri-implant bone loss in a well-maintained population. AB - The aim of the present study was (i) to investigate the relation between specific interleukin-1 (IL-1) gene polymorphisms and peri-implant bone loss at osseointegrated ITI(R) dental implants and (ii) to explore the association between these allelic variants of the IL-1 gene complex and peri-implant mucosal inflammation, in both smoking and non-smoking individuals. A sample of 90 consecutive Caucasian patients (aged 33-88 years), treated with at least one ITI implant participated in this retrospective investigation. Standardized periapical radiographs were taken after prosthetic rehabilitation (133.6 days, SD 136.9 days) and at the time of the re-examination, on average 5.6 years (SD 2.5 years) thereafter. The radiographs were analyzed by a calibrated examiner for changes in peri-implant bone levels. The examiner was blind with respect to clinical parameters and IL-1 status. The distance between the implant shoulder and the first visible bone-implant contact (DIB) at the respective time points were measured using a computerized method. The absolute bone level difference during the years of service (ABL) and the annual bone loss (DeltaBL/year) were calculated for all the implants. Percentages of full mouth bleeding on probing (BOP), as well as of BOP calculated separately for teeth and implants, were determined for all visits and averaged for the entire observation period. Out of the total patient sample, there were 14 heavy smokers (= 20 cigarettes/day), 14 moderate smokers (5-19 cigarettes/day), 23 previous smokers (smoking cessation > 5 years) and 39 non-smokers. Twenty-eight (31.11%) patients were IL-1 genotype positive. Upon stratification for smoking status, significant differences were found for the variables ABL (P < 0.04, U-test) and DeltaBL/year (P < 0.04, U test) between non-smokers and heavy smokers for the IL-1 genotype positive group but not for the IL-1 genotype negative group. Moreover, significant differences in ABL (P < 0.04, U-test) and DeltaBL/year (P < 0.04, U-test) were identified between former smokers and heavy smokers for the IL-1 genotype positive group. The differences in inflammatory parameters (BOP) did not reach statistical significance. This study suggests that in heavy cigarette smokers, carriage of a functionally significant IL-1 gene complex polymorphism is associated with an increased risk for peri-implant bone loss following prosthetic reconstruction and during the supportive periodontal care phase of the treatment. PMID- 12562361 TI - Advanced imaging: Magnetic resonance imaging in implant dentistry. AB - For accurate and safe placement of dental implants, and planning of associated surgery, a full assessment of the surgical anatomy of the site is necessary. Thus, it is highly desirable to have tomographic, sectional information available, to permit the implant to be aligned correctly. In recent years, X-ray computed tomography (CT) has become accepted as the gold standard in assessment, but the exposure to ionising radiation can be substantial. Artefacts due to dental restorations can also be significant, and some doubts may exist over the accuracy of reformatted CT. Magnetic resonance imaging (MRI) entails no exposure to ionising radiation, and allows direct acquisition of tomographic information in any desired plane. Sequential studies may be safely performed, allowing us a valuable insight into bone graft behaviour. Other than in a small number of cases, MRI may be safely used for presurgical assessments. Artefacts are few and in most cases localised. The surgical confidence from the sectional information gained is a significant step forward in the safe placement of dental implants. PMID- 12562362 TI - The microflora recovered from the outer-surfaces of the Frialit-2 implanto prosthetic connector. AB - The aim of the present investigation was to examine the outer-surface microbiota of the prosthetic connector of Frialit-2 implants, and to compare the microbial findings with the peri-implant parameters 2 years after functional loading. In 16 implant-treated patients (55.8 +/- 9.5 years) the outer-surface micro-organisms of the prosthetic connectors were determined in 32 Frialit-2 implants. The functional loading time of the prosthetic suprastructures was 24.1 +/- 13.8 months on average. After removal of the implant-supported restoration, microbial samples were obtained from the outer-surfaces of the Frialit-2 prosthetic connector. The microbial plaque samples were specified on CDC-blood agar as percentages of the total cultivable flora. Actinobacillus actinomycetemcomitans was semiquantitively determined on TSBV-agar in CFU/ml. The microbial plaque samples were dominated by Actinomyces israelii (68.8%), Eubacterium lentum (56.3%) and Veillonella parvula (43.8%) with proportions ranking between 3.9% (V. parvula) and 11.1% (A. israelii). The most frequently detected gram-negative microorganisms were Fusobacterium nucleatum (87.5%), Porphyromonas gingivalis (81.3%), and Peptostreptococcus micros (68.8%) with enhanced proportions for P. gingivalis (11.4%) and P. micros (11.4%). No statistical significant correlation could be established between the microbiota present on the outer-surfaces of the F2-connector and the peri-implant parameters examined. The outer-surface microflora recovered from the implanto-prosthetic-connector of Frialit-2 implants reveals a colonization with gram-positive bacteria and potentially harmful gram negative micro-organisms that were frequently detected, but present at low levels. After 2 years of restorative loading, the outer-surface microbial colonization is compatible with peri-implant soft tissue health. PMID- 12562363 TI - The jumping distance revisited: An experimental study in the dog. AB - Following tooth extraction, a socket often presents dimensions that may be considerably greater that the diameter of a conventional implant. The present experiment was performed to study the healing that occurred adjacent to implants placed in recipient sites with a wide marginal defect. Four Labrador dogs were used. In the right side of the mandible, four experimental sites were prepared to receive titanium implants [sandblasted, large-grit, acid-etched (SLA) surface]. Traditional implant installation (control) was performed in one site. In the remaining three sites (test), a step drill was used to widen the marginal 5 mm of the canal. Following placement of an implant in a test site, a circumferential gap about 1-1.25 mm wide and 5 mm deep was present lateral to the implant. A resorbable barrier membrane was used to cover the implant and the bone tissue of two sites, while one site was left uncovered. Four months following implant installation, block biopsies of each implant site were obtained and prepared for ground sectioning. After 4 months of healing, the large marginal defect had been filled with newly formed bone. The degree of bone-to-implant contact between the newly formed tissue and the SLA surface was at all test sites high and similar to that obtained at control sites. The placement of a barrier membrane following implant installation did not improve the outcome of healing. We conclude that a marginal defect wider than 1 mm may heal with new bone and a high degree of osseointegration to an implant designed with a SLA surface. PMID- 12562364 TI - Rat bone marrow cell response to titanium and titanium alloy with different surface roughness. AB - In general, cell response is affected by both chemical composition and surface roughness of implant materials. The aim of this study was to evaluate the effect of titanium (Ti) chemical composition and surface roughness on the response of rat bone marrow cells, examining cell attachment, cell proliferation, total protein content, alkaline phosphatase (ALP) activity, and bone-like nodule formation. Cells were cultured on both commercially pure titanium (cpTi) and titanium-6-aluminum-4-vanadium alloy (Ti-A) discs with four different average roughnesses (Ra). For attachment evaluation, cells were cultured for 2 h. After 14 days, cell proliferation, total protein content, and ALP activity were evaluated. Bone-like nodule formation was evaluated after 21 days. Data were compared by anova and Duncan's multiple range test when appropriate. Cell attachment and total protein content were affected by neither Ti chemical composition (P = 0.201, and P = 0.639, respectively) or surface roughness (P = 0.972, and P = 0.660, respectively). Proliferation, ALP activity, and bone-like nodule formation were affected only by Ti chemical composition (P = 0.0001, P = 0.064, and P = 0.0001, respectively). These results suggest that cpTi would optimize osteoblastic differentiation by rat bone marrow cells, including reduced cell proliferation, and increased ALP activity and bone-like nodule formation, while surface roughness, within the Ra parameters used, would not affect significantly the rat bone marrow cell response. PMID- 12562365 TI - Production of transforming growth factor beta1 and prostaglandin E2 by osteoblast like cells cultured on titanium surfaces blasted with TiO2 particles. AB - The surface roughness of an implant to which osteoblasts attach may influence endogenous expression of growth factor and cytokines at the implant-tissue interface, modulating the healing process and affecting the quality of bone formation. The present study, using cells derived from human mandibular bone, investigated the effect of varying roughness of titanium surfaces on production of transforming growth factor beta1 (TGF-beta1) and prostaglandin E2 (PGE2). The titanium surfaces were turned (control) and then roughened by blasting with 63-90 micro m, 106-180 micro m or 180-300 micro m TiO2 particles. The cells were cultured onto the surfaces till confluence was achieved. Fresh media were added in the presence or absence of 1,25-dihydroxyvitamin D3[1,25-(OH)2D3], the stimulator of osteogenic differentiation, and aliquots of conditioned cell media were used for assay 24 h later. Cellular morphology was determined by scanning electron microscopy. Cellular production of TGF-beta1 and PGE2 on each surface was assessed by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA), respectively, using commercially available kits. All blasted surfaces showed significantly higher production of TGF-beta1 than the turned surfaces (P < 0.05). In response to stimulation by 1,25-(OH)2D3 cellular production of TGF beta1, was also significantly greater (P < 0.05) on the blasted surfaces than on the turned one. The total amount of PGE2 in the conditioned media was higher than on the turned surfaces in the presence or absence of 1,25-(OH)2D3. There were no significant differences among the three blasted surfaces with respect to production of the local factors. However, we could not show a synergistic effect of surface roughness and vitamin D on the production of both TGF-beta1 and PGE2 using primary cell culture model. PMID- 12562366 TI - Ion implantation: surface treatment for improving the bone integration of titanium and Ti6Al4V dental implants. AB - Dental implants subjected to surface treatment have shown better bone integration than implants which have only been turned (machined). Three main types of treatment are presently available: the addition of material or coating, the removal of material, and surface modification. Ion implantation corresponds to the third approach. A histomorphometric study is made following the rabbit tibial bone placement of 88 commercial dental implants of pure titanium and Ti6AI4V subjected to surface treatment in the form of different ion implants (C+, CO+, N+, Ne+). Light microscopic, scanning electron microscopic (SEM), electron microsonde (EDS) and X-ray photoelectron spectroscopy (XPS) studies were made. The results indicate improved bone integration (expressed as percentage bone implant contact) in those specimens subjected to ion implantation versus the non treated controls, the difference being statistically significant for the groups treated with C+ and CO+. In these groups, XPS showed a Ti-O-C junction (bone implant interface) involving covalent type bonds, these being stronger and more stable than the ion-type bonds usually established between the titanium oxide and bone. PMID- 12562367 TI - Bone augmentation by means of a stiff occlusive titanium barrier. AB - It has already been shown that occlusive titanium barriers have osteoconductive properties. These barriers, however, cover only a limited surface area and have only been used in animal experiments. The aim of this study was to evaluate bone neogenesis under a pre-shaped titanium barrier placed over the top of the rabbit skull and the top of highly resorbed edentulous upper-jaw bone in patients. Computed tomography (CT) scans made it possible to pre-shape the titanium barrier according to individual bone shape in human experiments. On the rabbit skull, tissue augmentation of up to 6 mm 1 year after barrier placement was observed, while the original thickness of skull bone was on average between 1.5 and 2.5 mm. The bone, which remained histologically immature for 1 year, grew systematically along the titanium surface, illustrating its osteoconductivity. Even after removal of the barrier, on average, 75.3 and 59.4% of the newly created tissue volume was maintained after 3 and 9 months, respectively. Clinical observations on 10 consecutive patients showed that, in those (5/10) in which the barrier remained unexposed for several months, an increase of the jawbone height and width of up to 16 mm could be observed when the barrier was removed after 12-18 months. As in the rabbits at barrier removal, the bone demonstrated a limited degree of mineralization as ascertained from biopsies. This newly formed osteoid tissue allowed the insertion of 33 screw-shaped titanium implants which in most cases (30/33) successfully osseointegrated to support a fixed prosthesis. The surrounding marginal bone level remained stable even up to 5 years after implant placement. Both animal and clinical data demonstrate that guided bone neogenesis under a subperiosteally placed titanium barrier can reach large volumes. PMID- 12562368 TI - Anatomic site evaluation of the zygomatic bone for dental implant placement. AB - Thirty human zygomatic bone specimens (15 females mean age 81.60 +/- 11.38 years, 15 males, mean age 78.47 +/- 6.58 years) were examined by quantitative computed tomography and histomorphometry. The aim of the study was to assess the bone mineral density, the trabecular bone volume and the trabecular bone pattern factor. Moreover, the anterior-posterior and the medio-lateral dimensions and the estimated implant length within the zygomatic bone were determined. For quantitative computed tomography the specimens were scanned together with a bone mimicking anthropomorphic reference phantom. The bone mineral density was calculated for the specimens in the plane of the intended direction of the implant placement. Subsequently, with the sawing and grinding technique, the specimens were prepared in the same plane for histomorphometry. The trabecular bone mineral density was 369.95 +/- 188.80 mg/cm3 for the female and 398.94 +/- 99.11 mg/cm3 for the male specimens (P = 0.23). The male trabecular bone volume showed a value of 27.32 +/- 9.49%, while the female group reached a value of 19.99 +/- 7.60% (P = 0.23). The trabecular bone pattern factor was 1.2 x 10-2 +/- 1.28 mm-1 for the male and 1.02 +/- 0.96 mm-1 for the female specimens (P = 0.045). The study reveals that the zygomatic bone consists of trabecular bone with parameters that are unfavourable for implant placement. However, the success of implants placed in the zygomatic bone is secured by the employment of at least four cortical portions. PMID- 12562369 TI - Comparative analysis of collagen membranes for the treatment of implant dehiscence defects. AB - Guided bone regeneration (GBR) evolved from the concept of guided tissue regeneration (GTR) and has been used for reconstructing sites with bone deficiencies associated with dental implants. For GBR, the use of absorbable collagen membranes has been increasing, but, at present, scientific information on the use of collagen membranes for GBR is limited. This study was aimed to clinically and histomorphometrically compare two collagen membranes, Bio-Gide(R) and BioMend ExtendTM, for the treatment of implant dehiscence defects in eight mongrel dogs. Implant dehiscence defects were surgically created in edentulous ridges, followed by the placement of three endosseous implants bilaterally in the mandible. Each implant dehiscence defect was randomly assigned to one of three treatment groups: (1) control (no membrane), (2) porcine dermis collagen barrier (Bio-Gide) or (3) bovine tendon collagen barrier (BioMend Extend). Dogs were sacrificed at 4 and 16 weeks (four dogs each) after treatment. Histomorphometric analysis included percentage linear bone fill (LF), new bone-to-implant contact (BIC) and area of new bone fill (BF). The results of the study revealed no significant differences among groups for any parameter at 4 weeks. However, at 16 weeks, more LF, BIC, and BF were noted in the membrane-treated groups than controls. BioMend Extend-treated defects demonstrated significantly greater BIC than control (P < 0.05) at this time point. BIC at 16 weeks was significantly greater than 4-week BIC (P < 0.05). Membrane exposure occurred in 9 out of 15 sites examined, resulting in significantly less LF and BIC than the sites without membrane exposure (P < 0.05). The results of this study indicate that: (1) GBR treatment with collagen membranes may significantly enhance bone regeneration, manifested at late stage (16 weeks) of healing; and (2) space maintenance and membrane coverage were the two most important factors affecting GBR using bioabsorbable collagen membranes. PMID- 12562370 TI - Temperature changes induced by 809-nm GaAlAs laser at the implant-bone interface during simulated surface decontamination. AB - The aim of the study was to investigate temperature changes at the implant-bone interface during simulated implant surface decontamination with a 809-nm gallium aluminium-arsenid (GaAlAs) semiconductor laser. Stepped cylinder implants with a diameter of 3.8 mm and a length of 11 mm with two different surfaces (sand blasted and acid etched, and hydroxyapatite-coated) were inserted into bone blocks cut from freshly resected pig femurs. Access holes of 0.5 mm were drilled into the bone, to allow K-type thermocouples to contact periimplant bone in different parts of the cavity. An artificial periimplant bone defect provided access for laser irradiation in the coronal third. A 600-micrometer optic fiber was used at a distance of 0.5 mm from the implant surface. Power output varied between 0.5 and 2.5 W in the continuous wave mode. The bone block was placed into a 37 degrees C water bath in order to simulate in vivo thermal conductivity and diffusitivity of heat. Temperature elevations during irradiation were registered for a period of 120 s. In mean, the critical threshold of 47 degrees C was exceeded after 9.0 s at 2.5 W, 12.5 s at 2.0 W, 18.0 s at 1.5 W and 30.5 s at 1.0 W. Surface characteristics did not have a significant effect on temperature elevations. In an energy-dependent manner, implant surface decontamination with an 809-nm GaAlAs laser must be limited in time to allow the implant and bone to cool down. Clinical guidelines are presented to avoid tissue damage. PMID- 12562371 TI - Biomechanical stress in bone surrounding an implant under simulated chewing. AB - The concept of reducing nonaxial loading of dental implants has been widely regarded as the standard procedure. The aim of this study was to reveal the biomechanical stress distribution in supporting bone around an implant and a natural tooth under chewing function. Three-dimensional finite element models of the mandibular first molar and the titanium implant both with the mandible in the molar region were constructed. The directions of displacement constraints were determined according to the angles of the closing pathways of chopping type and grinding type chewing patterns. The tooth model showed smooth stress distribution in the supporting bone with low stress concentration around the neck of the tooth. The implant model showed stress concentration in the supporting bone around the neck of the implant, especially in the buccal area. The grinding type model of the implant showed higher tensile stress concentration than the chopping type model at the lingual neck of the implant. The results of this study suggested the importance of considering occlusion under chewing function for understanding the biomechanics of oral implants. PMID- 12562372 TI - Mandibular flexure and stress build-up in mandibular full-arch fixed prostheses supported by osseointegrated implants. AB - The biomechanical effect of mandibular functional flexure on stress build-up in implant-supported fixed restorations is discussed. The relative deformations and stress distributions in six different designs of implant-supported prosthetic systems (six or four implants, with or without distal cantilevers, cross-arch or midline-divided bar into two free-standing bridges) were analysed by a three dimensional finite element (FE) model of a human edentulous mandible. A significant amount of stress in the more distal implants and the superstructure at the symphysis arises as a consequence of mandible functional flexure. The analysis of the stress distributions generated by the different restorative patterns suggests that a division of the superstructure at the level of the symphysis significantly restores the natural functional flexure of the mandible. PMID- 12562373 TI - Effect of anxiety on the experience of pain in implant insertion. AB - Pain is a complex experience affected by such factors as stress, anxiety and cognitions. The purpose of this study was to examine the inter-relationship between anxiety and acute pain perception under an oral surgery procedure of implant insertion. The study population consisted of 60 dental patients (58% female, mean age 42 year), who were scheduled for implant insertion in a private clinic specializing in oral surgery. Patients were evaluated on three consecutive occasions: immediately preoperatively (T1), immediately post-operatively (T2), and at 4 weeks post-operative follow-up (T3). Patients were requested to complete questionnaires concerning their anxiety on each occasion and to indicate their subjective evaluations concerning pain (on visual analogue scales). Patient anxiety and pain evaluation were highest immediately before the surgical procedure (T1) with a significant decrease immediately afterwards (T2). The best predictor of the patient's pain evaluation at each time point was their state of anxiety at that time (T1: mean square = 7844.36, F = 16.26, P < 0.001; T2: mean square = 7652.74, F = 15.86, P < 0.001; T3: mean square = 5433.04, F = 8.99, P < 0.005). Pain experienced by patients in oral surgery is best predicted by their anxiety at each time point. Proper understanding of the variables that affect pain is important as they may produce emotional responses that could influence compliance. PMID- 12562374 TI - Patient satisfaction and quality of single-tooth restorations. AB - This retrospective study focuses on patient opinion and professionally assessed quality of single-tooth restorations of Branemark fixtures. Patients were treated by periodontists for fixture installation and/or abutment connection and by referring general practitioners for prosthetic rehabilitation. Seventy-eight consecutively treated patients were sent a questionnaire regarding aesthetics, phonetics and overall satisfaction. Forty-eight patients (52 fixtures) returned the questionnaire. Forty were able to attend a clinical examination by an independent examiner. The quality of 43 single implant crowns was evaluated according to the modified guidelines for assessment of quality and professional performance used for evaluation of design, fit, occlusion/articulation and aesthetics. Bone to implant level was assessed radiographically. Patients were very positive about aesthetics, phonetics, eating comfort and overall satisfaction. Nevertheless, 6/40 patients would not undergo the same treatment again, yet all of them would recommend it to others. The professionally rated quality of the crowns was perfect in 17 and acceptable in 25 cases. One crown needed modification to prevent future complications. Two crowns had already been replaced as a result of porcelain fracture; one restoration showed abrasion and one was damaged. With a mean loading time of 33 months (range: 3-89 months), none of the patients returning the questionnaire had encountered fixture loss. All except five fixtures showed bone remodelling at or above the first implant thread. It is concluded that single Branemark fixtures installed by periodontists and restored by general practitioners are of an acceptable clinical quality to satisfy the patient. PMID- 12562375 TI - Within-subject comparisons of maxillary fixed and removable implant prostheses: Patient satisfaction and choice of prosthesis. AB - Dental implants provide patients with restorative options for the edentulous maxilla. Both fixed and removable prostheses can be attached to the edentulous maxilla, but the efficacy of different designs has not been determined. In this two-session within-subject crossover trial we compared maxillary implant retained fixed prostheses with removable implant overdentures opposed by mandibular implant-supported overdentures. Sixteen patients, who had previously received mandibular implants, entered the study and received four to six maxillary implants. After dropouts as a consequence of a lack of osseointegration and general health problems, 13 remained. Of these, five received the removable prosthesis first and eight the fixed prosthesis. After 2 months, the prostheses were exchanged and the second was also worn for 2 months. Psychometric measurements of general satisfaction with the prostheses as well as comfort, ability to speak, stability, esthetics, ease of cleaning and occlusion were obtained once each prosthesis had been worn for 2 months. Chewing ability was assessed for seven types of food. Removable long-bar overdentures received significantly higher ratings of general satisfaction than fixed prostheses (P = 0.003). Patients also rated their ability to speak and ease of cleaning significantly better with the removable overdentures. Nine patients chose to keep the removable prosthesis and four preferred to keep the fixed prosthesis. The results suggest that maxillary removable overdentures on multiple implants may provide patients with better function than fixed prostheses. PMID- 12562376 TI - Accuracy of drilling guides for transfer from three-dimensional CT-based planning to placement of zygoma implants in human cadavers. AB - The accuracy of surgical drilling guides was assessed for placement of zygoma implants. Six zygoma fixtures of length 45 mm (Nobel Biocare, Goteborg, Sweden) were placed in three formalin-fixed human cadavers using surgical drilling guides. The fabrication of these custom-made drilling guides was based on three dimensional computerized tomography (3D-CT) data for the maxillary-zygomatic complex. The installation of the implants was simulated preoperatively using an adopted 3D-CT planning system. In addition, anatomical measurements of the zygomatic bone were performed on the 3D images. The preoperative CT images were then matched with postoperative ones in order to assess the deviation between the planned and installed implants. The angle between the planned and actually placed implants was < 3 degrees in four out of six cases. The largest deviation found at the exit point of one of the six implants was 2.7 mm. The present study showed that the use of surgical drilling guides should be encouraged for zygoma implant placement because of the lengths of the implants involved and the anatomical intricacies of the region. PMID- 12562377 TI - RSV bronchiolitis, gammadelta T cells and asthma: are they linked? PMID- 12562378 TI - Role of interleukin-12 in the regulation of CD4+ T cell apoptosis in a mouse model of asthma. AB - Allergic asthma, a chronic inflammatory disease of the airways, is characterized by the presence of T helper 2 cells and eosinophils in sputum, bronchoalveolar lavage, and mucosal biopsy specimens. Although the T helper 1-promoting cytokine, interleukin-12, is capable of inhibiting the T helper 2-driven asthma symptoms and bronchial responsiveness, the specific mechanisms underlying these interleukin-12 actions are unclear. The anti-allergic response to interleukin-12 is only partially dependent on interferon-gamma, which induces apoptosis by enhancing expression of Fas antigen. We therefore investigated in vivo whether the anti-allergic action of interleukin-12 is mediated through induction of apoptosis. C57BL/6 mice immunized to ovalbumin by intraperitoneal injection were challenged three times with an ovalbumin aerosol every second day for 7 days. Recombinant interleukin-12 was administered intravenously after the final challenge. After the last ovalbumin challenge, mice were examined for effects of interleukin-12 on inflammatory cell infiltration and apoptosis in the lung as detected by terminal deoxynucleotidyl transferase-mediated deoxyribonucleoside triphosphate nick end-labelling. Administration of interleukin-12 reduced ovalbumin-induced pulmonary eosinophilia (P < 0.01) and CD4+ T cell infiltration (P < 0.01). Moreover, treatment with interleukin-12 shortly after ovalbumin inhalation resulted in both increased interferon-gamma production (P < 0.01) and enhanced apoptosis of CD4+ T cells in allergic airway infiltrates (P < 0.05). These results suggest that the beneficial effects of interleukin-12 in asthma may include enhancement of apoptosis of CD4+ T cells in airways. PMID- 12562379 TI - Interleukin 10 induced augmentation of delayed-type hypersensitivity (DTH) enhances Mycobacterium bovis bacillus Calmette-Guerin (BCG) mediated antitumour activity. AB - Intravesical BCG therapy is effective in the treatment of superficial bladder cancer. Both clinical and experimental results suggest a role for cytokines and delayed-type hypersensitivity (DTH) in BCG-induced antitumour immunity. We characterized the modulatory effects of BCG on bladder cytokine expression and determined the relationship between DTH and BCG antitumour activity. The bladders of mice were instilled with BCG through a catheter. Bladder tissue RNA and urine were collected for evaluation of cytokine expression using reverse transcriptase polymerase chain reaction (RT-PCR) and/or ELISA. IFN-gamma and TNF-alpha, the two major cytokines associated with DTH, were efficiently induced by BCG. IL10, an important down-regulator of DTH, was also induced by BCG. Constitutive levels of IL4 and IL5 were observed, but neither IL4 nor IL5 were modulated by BCG. Similar results were observed in the kinetic analysis of urinary cytokines in patients after intravesical BCG therapy. Production of Th1 (T helper type 1) cytokines (IFN-gamma, IL2 and IL12) preceded that of the Th2 (T helper type 2) cytokine IL10. A tendency toward higher ratios of IFN-gamma versus IL10 for BCG responders also was observed. In animal studies the absence of IL10 abrogated either by antibody inhibition or the use of genetically modified, IL10 deficient (IL10-/-) mice resulted in enhanced DTH responses. Under conditions of enhanced DTH, a significant enhancement in antitumour activity was observed. These data demonstrate that DTH and its associated mononuclear infiltration and cytokine production are important to the antitumour activity of intravesical BCG therapy, and suggest that effects to diminish IL10 production may have therapeutic value. PMID- 12562380 TI - Endogenous glucocorticoids attenuate Shiga toxin-2-induced toxicity in a mouse model of haemolytic uraemic syndrome. AB - The concept that during an immune challenge the release of glucocorticoids (GC) provides feedback inhibition on evolving immune responses has been drawn primarily from studies of autoimmune and/or inflammatory processes in animal models. The epidemic form of haemolytic uraemic syndrome (HUS) occurs secondary to infection with Gram-negative bacteria that produce Shiga toxin (Stx). Although Stx binding to the specific receptors present on renal tissue is the primary pathogenic mechanism, inflammatory or immune interactions are necessary for the development of the complete form of HUS. The aim of this study was to investigate the influence of endogenous GC on Stx-toxicity in a mouse model. Stx2 was injected into GC-deprived mice and survival rate, renal damage and serum urea levels were evaluated. Plasma corticosterone and cytosolic GC receptor (GR) concentration were also determined at multiple intervals post-Stx2 treatment. Higher sensitivity to Stx2 was observed in mice lacking endogenous GC, evidenced by an increase in mortality rates, circulating urea levels and renal histological damage. Moreover, Stx2 injection was associated with a transient but significant rise in corticosterone secretion. Interestingly, 24 h after Stx inoculation significant increases in total GR were detected in circulating neutrophils. These results indicate that interactions between the neuroendocrine and immune systems can modulate the level of damage significantly during a bacterial infection. PMID- 12562381 TI - Enhanced chemokine response in experimental acute Escherichia coli pyelonephritis in IL-1beta-deficient mice. AB - The aim of the present study was to investigate the effects of IL-1beta and Escherichia coli on the expression and secretion of MIP-2, the mouse equivalent to human IL-8, MCP-1 and RANTES in the kidneys of mice with acute pyelonephritis. Female Bki NMRI, as well as IL-1beta deficient mice and their wild-type littermates, were transurethrally infected with either E. coli CFT 073 or injected with NaCl 0.9% (w/v) and thereafter obstructed for 6 h. The Bki NMRI mice were killed at 0, 24, 48 h and 6 days and the IL-1beta-deficient mice at 48 h. Chemokine mRNA and protein levels peaked at 24 h for the tested chemokines with the mRNA expression localized in the tubular epithelial cells and for MIP-2 also in neutrophils. Obstruction per se, also induced a chemokine expression similar to E. coli infection although at a lower level. Interestingly, MIP-2 levels were higher in the IL-1beta deficient mice as compared with the wild-type littermates. Likewise, the inflammatory changes were more frequent and, when present, more widespread in the IL-1beta-deficient mice than in the wild-type mice. Stimulation of a human renal tubular epithelial cell line (HREC), A498 and of primary human mesangial cells (HMC) with the same bacterial antigen depicted gene expression of the same chemokines. A rapid release of IL-8 and MCP-1 was observed from both cell types. RANTES response was delayed both in the HREC and the HMC. We conclude that acute E. coli pyelonephritis induces a MIP-2/IL-8, MCP 1 and RANTES expression and secretion localized primarily to the epithelial cells and that this production is confirmed after in vitro stimulation with the same bacterial antigen of human epithelial and mesangial cells. Blockade of induction of chemokine response may thus be an attractive target for possible therapeutic intervention. PMID- 12562382 TI - Dendritic cells infected with adenovirus expressing the thyrotrophin receptor induce Graves' hyperthyroidism in BALB/c mice. AB - Dendritic cells (DCs) are the most potent antigen-presenting cells and a prerequisite for the initiation of primary immune response. This study was performed to investigate the contribution of DCs to the initiation of Graves' hyperthyroidism, an organ-specific autoimmune disease in which the thyrotrophin receptor (TSHR) is the major autoantigen. DCs were prepared from bone marrow precursor cells of BALB/c mice by culturing with granulocyte macrophage-colony stimulating factor and interleukin-4. Subcutaneous injections of DCs infected with recombinant adenovirus expressing the TSHR (but not beta-galactosidase) in syngeneic female mice induced Graves'-like hyperthyroidism (8 and 35% of mice after two and three injections, respectively) characterized by stimulating TSHR antibodies, elevated serum thyroxine levels and diffuse hyperplasitc goiter. TSHR antibodies determined by ELISA were of both IgG1 (Th2-type) and IgG2a (Th1-type) subclasses, and splenocytes from immunized mice secreted interferon-gamma (a Th1 cytokine), not interleukin-4 (a Th2 cytokine), in response to TSHR antigen. Surprisingly, IFN-gamma secretion, and induction of antibodies and disease were almost completely suppressed by co-administration of alum/pertussis toxin, a Th2 dominant adjuvant, whereas polyriboinosinic polyribocytidylic acid, a Th1 inducer, enhanced splenocyte secretion of IFN-gamma without changing disease incidence. These observations demonstrate that DCs efficiently present the TSHR to naive T cells to induce TSHR antibodies and Graves'-like hyperthyroidism in mice. In addition, our results challenge the previous concept of Th2 dominance in Graves' hyperthyroidism and provide support for the role of Th1 immune response in disease pathogenesis. PMID- 12562383 TI - Antigen inhibition of collagen-induced arthritis is associated with up-regulation of IL-4 mRNA and induction of Ox40 on T cells in draining lymph nodes. AB - The addition of a foreign antigen to an inoculum completely inhibits the development of collagen-induced arthritis (CIA). However, the mechanism of this phenomenon, antigen -inhibition, is incompletely understood. Previous studies have demonstrated that the inhibition of arthritis is not mediated through suppression of the antibody response to cartilage antigens. In this paper we investigated cytokine mRNA levels in lymph nodes cells recovered 3, 7 or 16 days from animals immunized with either collagen II in IFA or OVA + collagen II in IFA. At day 7, but not at other time-points, IL-4 mRNA was up-regulated in the lymph nodes of OVA-inhibited non-arthritic animals compared to control animals which all developed arthritis. No significant differences between the two groups could be detected when expression of IFN-gamma, IL-2, TNF-alpha, IL-1beta or IL 10 mRNA was analysed. Flow cytometry analysis of draining lymph node cells demonstrated that the T cell marker Ox40 was up-regulated in the OVA-inhibited group. Our results indicate that the complete inhibition of CIA caused by addition of OVA to the collagen II inoculum is due to the presence of a TH2 environment resulting from an increased production of IL-4 mRNA and a parallel increase in Ox40+ T cells. PMID- 12562384 TI - Differential cytokine expression of human retinal pigment epithelial cells in response to stimulation by C5a. AB - Human retinal pigment epithelial (RPE) cells form part of the blood-retina barrier where they potentially can regulate leucocyte function. RPE cells are known to secrete several cytokines in response to stimulation by other cytokines. Anaphylatoxin C5a, a potent inflammatory mediator produced during complement activation, binds to G-protein coupled C5a receptors (C5aR) on monocytes/macrophages and releases various cytokines from the cells. We previously reported that the human RPE cell line ARPE-19 possesses C5aR and expresses IL-8 mRNA in response to C5a stimulation. In this study, we used a primary human RPE cell line (RPE43) and found that C5a induces increased expression of IL-1beta, IL-6, MCP-1 and GM-CSF mRNAs as well as IL-8 mRNA. ARPE 19 cells showed similar increases in the same cytokines. Interestingly, the kinetics of expression of the various cytokines differed. These results provide further evidence that C5a stimulation of RPE cells may play a role in regulating leucocyte function during ocular inflammation in which there is complement activation. PMID- 12562385 TI - Complement resistance of human carcinoma cells depends on membrane regulatory proteins, protein kinases and sialic acid. AB - Nucleated cells employ several strategies to evade killing by homologous complement. We studied complement resistance in the human carcinoma cell lines (CA) T47D (mammary), SKOV3 (ovarian), and PC-3 (prostate) with emphasis on the following mechanisms of defense: 1. Expression and shedding of the membrane complement regulatory proteins (mCRP) CD46, CD55 and CD59; 2. Resistance based on protein phosphorylation; 3. Cell surface expression of sialic acid residues; 4. Desensitization to complement upon exposure to sublytic complement doses. Anti mCRP antibody blocking experiments demonstrated that CD59 is the main mCRP protecting these CA from complement. Soluble CD59 was also found in supernates of PC-3> SKOV3 > T47D cells. Second, inhibitors of PKC, PKA and MEK sensitized the CA to lysis, thus implicating these protein kinases in CA complement resistance. Third, removal of sialic acid residues with neuraminidase also sensitized CA to lysis. Finally, exposure of CA to sublytic doses of complement conferred on them enhanced resistance to lytic complement doses in a PKC-dependent process. Combined treatment of CA with anti-CD59 antibodies, PD98059 (a MEK inhibitor) and neuraminidase produced a large enhancement in CA sensitivity to complement. Our results show that CD59 and sialic acid residues present on the cell surface, and intracellular processes involving protein phosphorylation act additively to secure CA resistance to complement-mediated lysis. Therefore, the effectiveness of antibody- and complement-based cancer immunotherapy will markedly improve by suppression of the various complement resistance mechanisms. PMID- 12562386 TI - Atorvastatin suppresses interferon-gamma -induced neopterin formation and tryptophan degradation in human peripheral blood mononuclear cells and in monocytic cell lines. AB - Inhibitors of 3-hydroxy-3methylglutaryl-co-enzyme A (HMG-CoA) reductase, so called statins, are used in medical practice because of their lipid-lowering effect and to reduce the risk of coronary heart disease. Recent findings indicate that statins also have anti-inflammatory properties and can modulate the immune response. In vitro, we investigated the effect of atorvastatin on the T cell/macrophage system in peripheral blood mononuclear cells (PBMC) and in the human monocytic cell lines THP-1 and MonoMac6. We monitored neopterin production and tryptophan degradation in PBMC after treatment with 10 micro m and 100 micro m atorvastatin in the presence or absence of 100 U/ml IFN-gamma, 10 micro g/ml phytohaemagglutinin (PHA) or 10 micro g/ml concanavalin A (ConA) and in monocytic cell lines THP-1 and MonoMac6 with or without stimulation with 100 U/ml IFN-gamma or 10 ng/ml to 1 micro g/ml lipopolysaccharide (LPS). In stimulated PBMC 100 micro m atorvastatin inhibited neopterin formation and tryptophan degradation completely, whereas 10 micro m atorvastatin was only partially effective. Also in monocytic cell lines THP-1 and MonoMac6, atorvastatin was able to suppress IFN gamma- and LPS-induced formation of neopterin and degradation of tryptophan. Our data from PBMC agree well with previous investigations that statins inhibit T cell activation within the cellular immune response. In addition we demonstrate that atorvastatin directly inhibits IFN-gamma-mediated pathways in monocytic cells, suggesting that both immunoreactivity of T cells and of monocyte-derived macrophages are down-regulated by this statin. PMID- 12562387 TI - Benzylpenicillin differentially conjugates to IFN-gamma, TNF-alpha, IL-1beta, IL 4 and IL-13 but selectively reduces IFN-gamma activity. AB - It is known that beta-lactam antibiotics can conjugate to lysine and histidine residues on proteins via the carbonyl group of the opened beta-lactam ring. However, it is not known which proteins these drugs target and there is little work addressing whether conjugation is preferential for some proteins over others or if conjugation has functional consequences for the protein. We have previously shown that the beta-lactam antibiotic benzylpenicillin (BP) conjugates to IFN gamma and reduces its activity. This interaction demonstrates selectivity, as BP does not bind to IL-4. Here, we extend our study to include other Th1 and Th2 cell-associated cytokines and two cytokines associated with inflammatory responses. We demonstrate by Western blotting that BP also conjugates to IL 1beta, IL-2, IL-5, IL-13 and TNF-alpha but not to IL-10. Densitometric analysis of leading cytokine bands on blots revealed that IFN-gamma always gave more intense BP-positive bands than any other cytokine analysed. Cytokines pre incubated with BP at 37 degrees C in a protein-containing, serum-free medium were assayed for their biological activity. By in vitro bioassay, BP inhibited the ability of IFN-gamma but not IL-1beta or TNF-alpha to induce CD54 expression on epithelial cells. In addition, BP did not affect IL-4 or IL-13 inhibition of mast cell proliferation. When the pre-incubation temperature was reduced to 4 degrees C, BP did not conjugate to IFN-gamma or modulate its activity. BP retained its inhibitory effect on IFN-gamma activity when 20% FCS was added to the pre incubation medium. In conclusion, BP conjugates to some cytokines but not others and this does not appear to be related to primary protein structure. Furthermore, of the cytokines studied, conjugation only to IFN-gamma is accompanied by inhibition of activity. This phenomenon is temperature dependent and occurs in the presence of serum. These findings provide further evidence for differential, direct drug-cytokine interactions. Such interactions may have therapeutic implications in terms of targeting cytokines to regulate their activity. PMID- 12562388 TI - High positive frequency of antibodies to metallothionein and heat shock protein 70 in sera of patients with metal allergy. AB - Two principal types of stress protein, heat shock proteins (hsps) and metallothionein (MT), are induced in cells responding to a variety of stresses. They play an important role in protecting cells from these stresses. However, many reports indicate that antibodies to hsps are present in human serum and are associated with several autoimmunity diseases. Metals, which are commonly allergenic to humans, induce both MT and hsp70 (one of the hsps family). Until now, there has been no report of any antibody to MT in human serum. In the present study, serum samples from healthy controls (Group I), and patients suffering from atopic dermatitis without (Group II) or with (Group III) metal allergy, were measured for antibodies to MT and hsp70, using an enzyme-linked immunosorbent assay (ELISA). Metal allergy was confirmed by patch testing. We first found that antibody to MT exists in human serum. We also found a high positive frequency of antibody to MT (51.3%) and to hsp70 (43.6%) in the sera of Group III, compared to those of Group I (3.8% and 5.1%) or Group II (6.4% and 5.1%). Furthermore, there was a strong positive correlation between antibody to MT and antibody to hsp70 in Group III (P = 0.0013), but not in Group I and Group II. Our results indicate that antibody to MT exists in human serum, as do antibodies to hsps, and suggest that elevated levels of MT and hsp70 antibodies are associated with metal allergy in atopic patients. PMID- 12562389 TI - Molecular characterization of homozygous hereditary factor I deficiency. AB - We have studied the molecular basis of factor I (fI) deficiency in two Brazilian sisters from a consanguineous family. By reverse transcription-polymerase chain reaction we observed that all fI cDNA amplified products from one sister had the same size as those of normal cDNA, however, they were significantly less intense. Sequencing analysis of subcloned cDNA revealed a dinucleotide insertion (AT) between positions 1204 and 1205 in the 11th exon that creates a stop codon 13 bp downstream of the insertion site. Genomic DNA sequencing and heteroduplex analysis confirmed that both probands are homozygous for this mutation, whereas their parents are heterozygous. The stop codon and the diminished amounts of fI cDNA could indicate increased fI mRNA instability, perhaps due to a mechanism of nonsense-mediated decay. This hypothesis is consistent with our observation that treatment with the translation inhibitor cycloheximide stabilized fI mRNA expression in proband's fibroblasts. PMID- 12562390 TI - Gamma/delta T cell subsets in patients with active Mycobacterium tuberculosis infection and tuberculin anergy. AB - Earlier data suggest that gamma/delta T cells may play an important role in the immune response to Mycobacterium tuberculosis. The aim of this study was to determine the percentage of different gamma/delta subsets in peripheral blood of active tuberculosis patients with a positive or negative tuberculin reaction. Thirty-eight patients infected with M. tuberculosis and 22 healthy controls were included in the study. Venous blood was taken before starting antimycobacterial treatment. Lymphocytes were reacted with monoclonal antibodies specific for different gamma/delta V chains (Vdelta1, Vdelta2, Vgamma9 and Vgamma4). The results were analysed in the context of tuberculin reactivity and X-ray findings. Our results revealed a selective loss of Vgamma9/Vdelta2 T cells in the peripheral blood of tuberculin-negative patients with active tuberculosis compared to healthy controls, while the ratio of Vgamma9/Vdelta2 T cells in the peripheral blood of patients with a positive skin test did not differ from that of healthy controls. These findings demonstrate a relationship between the loss of the major M. tuberculosis-reactive subset of gammadelta T cells and the absence of tuberculin reactivity. The data are consistent with the hypothesis that gammadelta T cells play a role in the protective immune response to M. tuberculosis infection. PMID- 12562391 TI - Influence of Mycobacterium tuberculosis on differential activation of helper T cells. AB - Host defence against tuberculosis infection involves T-lymphocyte mediated cellular immune responses. In this study we assessed T-cell activation by studying the early signal transduction events and production of cytokines by human CD4+ T-cells. The study constituted of five groups of subjects: (a) untreated acid fast bacilli (AFB)+ve TB patients who have not started anti tuberculosis therapy (ATT) [New]; (b) patients who have taken ATT for two months [2T]; (c) patients who have taken ATT for six months [6T]; (d) mantoux positive healthy controls [T+ve]; (e) mantoux negative healthy controls [T-ve]. We found that mantoux positive healthy controls produced significantly higher levels of IP3, intracellular Ca2+ and presented increased PKC activity when CD4+ T-cells were stimulated with M. tuberculosis H37Rv cell lysate as compared to mantoux negative controls. Furthermore, decreased expression of CD54 (ICAM-1) and reduced [Ca2+]i were seen in TB patients as compared to T+ve healthy controls. TB patients showed significantly lower levels of IL-2 and IFNgamma and higher levels of IL-4 as compared to normal healthy controls, suggesting a diminished Th1 response. Thus, the reciprocal changes in cytokines, reduced [Ca2+]i levels, and CD54 expression in patients imply phenotype shifting of Th precursors to Th2 type in TB patients. PMID- 12562392 TI - Chlamydia trachomatis heat shock protein-60 induced interferon-gamma and interleukin-10 production in infertile women. AB - Chlamydia trachomatis-associated tubal factor infertility (TFI) involves enhanced humoral and cell-mediated immune response to the chlamydial 60 kDa heat shock protein (CHSP60). We evaluated the role of CHSP60-induced immune response in TFI by studying lymphocyte proliferation and cytokine (interferon (IFN)-gamma, interleukin (IL)-12 and IL-10) secretion in response to C. trachomatis elementary body (EB) and CHSP60 antigens in 57 women with TFI and in 76 women with other causes of infertility. Positive proliferative response of PBMC to CHSP60 was more common in the TFI group (20/57; 36%) than in the other groups (17/76; 22%) although the frequency or the median responses did not differ significantly (1.6, range 0.2-22.1 versus 1.4; 0.2-24.4). C. trachomatis EB induced significantly higher IFN-gamma and lower IL-10 secretion in the TFI group compared to the other groups. The EB and CHSP60 induced IL-12 secretion was similar in all study groups and correlated with IFN-gamma secretion in the other but not in the TFI group. The lack of correlation between EB-induced IL-12 and IFN-gamma production and simultaneously found prominent IL-10 secretion in response to CHSP60 in the TFI group suggests that the CHSP60 may have a specific role in regulating the immune reactions during chlamydial infection and may consequently contribute to the immunopathogenesis of TFI. PMID- 12562393 TI - Decreases in plasma TNF-alpha level and IFN-gamma mRNA level in peripheral blood mononuclear cells (PBMC) and an increase in IL-2 mRNA level in PBMC are associated with effective highly active antiretroviral therapy in HIV-infected patients. AB - In this study, we investigated the cytokine profiles of 14 treatment-naive HIV infected patients on the initiation of highly active antiretroviral therapy (HAART). At baseline, plasma levels of TNF-alpha and its mRNA in peripheral blood mononuclear cells (PBMC) were highest in the most severely immunocompromised patients (<200 CD4+ cells/mm3). After 12 months of HAART, the virus was undetectable in the plasma of all patients (<200 copies/ml), and median CD4 T cell counts had increased (+164 cells/mm3). We also observed a gradual decrease in the number of proviral DNA copies in PBMC and in immune activation, with lower levels of IFN-gamma mRNA in PBMC associated with weaker activation of CD8+ T cells and lower levels of plasma TNF-alpha. IL-2 mRNA levels in PBMC were found to increase in parallel. The decrease in TNF-alpha and IFN-gamma levels and the increase in IL-2 production appear to be correlated with the efficacy of HAART in naive immunocompromised HIV-infected individuals. PMID- 12562394 TI - Respiratory syncytial virus infection suppresses IFN-gamma production of gammadelta T cells. AB - The immunological mechanisms by which respiratory syncytial virus (RSV) contributes to the development of asthma are poorly understood. gammadelta T cells are important in mucosal defence, and may contribute to the establishment of primary immune responses by producing cytokines early during respiratory infections. Thus, we used flow cytometry and intracellular cytokine staining to investigate the expression of interferon (IFN)-gamma and interleukin (IL)-4 by mitogen-stimulated gammadelta T cells from the peripheral blood of 15 hospitalized infants with RSV bronchiolitis, seven rotavirus-infected infants and eight normal controls. gammadelta T cells from RSV-infected infants had a lower proportion of IFN-gamma-producing cells (median, 4.00%; range, 0.58-6.60%) and a slightly but significantly higher proportion of IL-4-producing cells (median, 0.40%; range, 0.13-2.76%) than rotavirus-infected infants (median, 32.10%; range, 14.43-61.21%; P < 0.01, median, 0.00%; range, 0.00-0.00%; P < 0.05) in the acute phase. By contrast, differences in cytokine production by total CD3+ T cells did not differ significantly between patient groups. Thus, reduced IFN-gamma production by gammadelta T cells in the peripheral blood of RSV-infected infants is accompanied by increased Th2 cytokine production during the acute phase of disease. At follow-up, eight children had recurrent episodes of wheezing. The frequencies of IFN-gamma-producing gammadelta T cells were significantly lower in patients who developed recurrent wheezing (median, 0.65%; range, 0.02-1.75%) than in patients without recurrent wheezing (median, 6.90%; range, 5.25-10.98%; P < 0.005). Cytokine production by gammadelta T cells may therefore be important in the pathogenesis of acute RSV disease, and play a part in the development of recurrent childhood wheezing after bronchilolitis. PMID- 12562395 TI - Local immune responses and systemic cytokine responses in zoster: relationship to the development of postherpetic neuralgia. AB - Varicella zoster virus (VZV) causes varicella (chickenpox) as the primary infection and zoster (shingles) on reactivation from latency, often many years later. One of the most common and most severe sequela of zoster is postherpetic neuralgia (PHN). Apart from age, factors which predispose towards PHN are unknown. In the present study, the concentration of a variety of Th1 and Th2 cytokines in the serum of 30 zoster patients at the time of the acute disease were correlated with the subsequent development of PHN in nine of these patients, but no association was found. In addition, although some cytokines such as IFN gamma, IL-6 and IL-8 were slightly raised in the zoster group compared with a group of normal healthy subjects of a similar age distribution, these differences only verged on significance. Antibody titres to VZV were raised in the zoster group compared with the controls but these did not differ between the patients who developed PHN and those who did not. Biopsies of zoster lesions were collected from nine patients. There were significantly fewer infiltrating lymphocytes in the lesions of the three patients who subsequently developed PHN compared with the six who did not, although the expression of the neuropeptide, substance P, did not differ between the two groups. It is possible that the poor inflammatory response at the time of the acute zoster may result in less effective containment of the VZV and more damage in the dermatome, thus contributing to the persistence of the neuralgia. PMID- 12562396 TI - Monocyte-derived cytokines in multiple sclerosis. AB - MS is an inflammatory, presumably autoimmune, disease mediated by the activation of T cells, B cells and monocytes (MO). Inflammation is thought to occur early during the relapsing-remitting phase of MS (RRMS), whereas in the later phases of MS such as secondary progressive MS (SPMS), inflammation tends to diminish. Our objective was to compare the types and amounts of proinflammatory and regulatory cytokines produced by MO from relapsing-remitting patients with or without treatment with IFN-beta (RRMS+ therapy, RRMS- therapy), respectively, from secondary progressive patients (SPMS) and from healthy controls (HC). MO were isolated by a density-gradient technique and three different techniques (RNase protection assay, ELISA and intracellular cytokine staining) were used to assess cytokine levels. An increase in IL6, IL12 and TNF-alpha was observed by all three methods for RRMS- therapy and for SPMS patients compared to HC and RRMS+ therapy patients. We conclude that proinflammatory and regulatory monokines can be derived from MO of MS patients and that these levels are modulated by IFN-beta therapy. Although it is believed that inflammation tends to diminish in SPMS patients, our data show that inflammatory cytokines continue to be released at high levels, suggesting that IFN-beta or IL10 treatment may be beneficial for this group. PMID- 12562397 TI - Flow cytometric determination of glucocorticoid receptor (GCR) expression in lymphocyte subpopulations: lower quantity of GCR in patients with post-traumatic stress disorder (PTSD). AB - Assessment of the intracellular glucocorticoid receptor (GCR) level may be useful in monitoring functional disturbances of the hypothalamic-pituitary adrenocortical axis or effects of prolonged steroid therapy. Cytosolic ligand binding assays have recently been supplemented by flow cytometric determination of receptor expression in individual cells. A method based on multiparametric analysis of whole blood by simultaneous labelling of intracellular GCRs and surface markers of lymphocyte subsets is described. We examined 25 healthy male volunteers and 35 age- and sex-matched post-traumatic stress disorder (PTSD) patients within 8 years from traumatic event. PTSD patients had a lower relative quantity of GCR in all lymphocyte populations tested as compared with healthy volunteers. NK cells of both groups showed higher expression of GCR than other lymphocyte subsets. In PTSD patients, the expression of GCR in B lymphocytes was also higher than in T cell. Although serum cortisol level was lower in PTSD patients, there was no correlation between cortisol level and GCR expression. Multiparameter flow cytometric determination of GCR expression in lymphocyte subpopulations may provide a useful tool for monitoring immunoregulatory action of glucocorticoids. PMID- 12562398 TI - Antigens shared by malignant plasma cells and normal B cells may be involved in graft versus myeloma. AB - Cytotoxic T cells play an important role in graft-versus-host-disease (GvHD) and graft-versus-leukaemia/myeloma, which may occur in patients treated with an allogeneic stem cell transplantation (ASCT). Here, we describe the selection of a myeloma reactive CD4+ cytotoxic T cell-line (CTL) and two CD4+ clones from this CTL. The CTL was generated from the blood from a patient with multiple myeloma (MM) with graft versus myeloma/GvHD, following an ASCT. The CTL was stimulated using irradiated peripheral blood mononuclear cells and EBV transformed B cells from the myeloma patient (EBVp), both of which were obtained prior to ASCT. Both the CTL and the two T cell clones specifically lysed EBVp and secreted IFN-gamma after coculture with EBVp and autologous myeloma tumour cells in a class II restricted fashion. These results show that myeloma tumour cells and autologous B cells present a common polymorphic peptide that functions as a target for graft derived cytotoxic T cells. Identification of these proteins will give insight into the relationship between graft versus myeloma (GvM) and GvHD and may provide immunotherapeutical targets in the treatment of MM. PMID- 12562399 TI - Autoimmune diseases in vitiligo: do anti-nuclear antibodies decrease thyroid volume? AB - An increased prevalence of autoimmune thyroiditis (AT) in vitiligo patients is well known. The aim of this study was firstly, to evaluate the clinical course of patients with both vitiligo and AT and secondly, to identify additional autoimmune disorders affecting the thyroid gland in a large cohort of vitiligo patients. We analysed a study group of 106 vitiligo patients and 38 controls. A detailed thyroid examination including sonography was performed in all study participants. In addition, the study participants were HLA typed and screened for various autoimmune disorders. AT was significantly more frequent in vitiligo patients than in controls (21%versus 3%; P < 0.01). In 12 of the 22 patients with AT, vitiligo was the initial disease preceding AT by 4-35 years. In the other 10 patients with AT, both vitiligo and AT were diagnosed within one year. There were two individuals with diabetes mellitus type 1 and a single patient with Addison's disease. Anti-nuclear antibody (ANA), anti-smooth muscle cell antibody, and parietal cell antibody levels occurred with a similar frequency in patients and controls. In all vitiligo patients with both elevated ANA levels and AT (n = 6), the atrophic but not the goitrous variant was diagnosed. These vitiligo patients with both AT and elevated ANA levels had a significantly smaller thyroid volume compared to the vitiligo patients with AT whose ANA levels were normal (6.7 +/- 4.5 ml versus 13.4 +/- 9.1 ml, respectively; P < 0.05). The same was found in the entire study group: Thyroid volume of all vitiligo patients (with or without concomitant AT) was significantly smaller in the presence of ANA (6.9 +/- 5.3 versus 10.5 +/- 5.9 ml, respectively; P < 0.05). However, this phenomenon was not observed in the control group. There was a trend for a decreased frequency of HLA DR3 (6.7%versus 23%) in our study group, but after correction for the number of comparisons, no HLA-allele was statistically significant associated neither with vitiligo nor with multiple autoimmune diseases in our patient sample. Our findings suggest that AT is the most frequent autoimmune disease associated with vitiligo. In our patients, AT presented simultaneously or after the onset of vitiligo but not before. Elevated ANA levels were associated with the atrophic variant of AT and may affect the volume of the thyroid gland, and there was no statistically significant association with the HLA system. PMID- 12562400 TI - High circulating levels of biologically inactive IL-6/SIL-6 receptor complexes in systemic juvenile idiopathic arthritis: evidence for serum factors interfering with the binding to gp130. AB - We previously demonstrated that high levels of IL-6/sIL-6R complexes are present in sera of patients with systemic juvenile idiopathic arthritis (s-JIA) and that the amount of IL-6 estimated in the IL-6/sIL-6R complexes is markedly higher than that measured by the B9 assay. Here, we show that two additional bioassays, employing human myeloma XG-1 cells and human hepatoma Hep3B cells, detected serum IL-6 levels similar to those measured by the B9 assay and approximately 10-fold lower than the IL-6 levels estimated to be present in the IL-6/sIL-6R complex. Using an assay for the measurement of the amount of circulating IL-6 complexed with the sIL-6R and available for binding to gp130 (gp130 binding activity), we show that the IL-6/gp130 binding activity is similar to that detected by the bioassays and again significantly lower than that estimated to be present in the IL-6/sIL-6R complex. Addition of recombinant human IL-6 (rhIL-6) to sera of patients or controls results in a markedly lower increase in the gp130 binding activity in patients than in controls. Moreover, sera from s-JIA patients inhibited in a dose dependent manner the gp130 binding activity assay. These results show that sera from patients with s-JIA contain a factor, or factors, that inhibit(s) the binding of the IL-6/sIL-6R complex to gp130. This inhibitory activity does not appear to be due to soluble gp130, C-reactive protein or autoantibodies to IL-6. PMID- 12562401 TI - Human IgG Fc-binding phage antibodies constructed from synovial fluid CD38+ B cells of patients with rheumatoid arthritis show the imprints of an antigen dependent process of somatic hypermutation and clonal selection. AB - The persistent presence of rheumatoid factors (RFs) in the circulation is a characteristic phenomenon in patients with rheumatoid arthritis (RA). Recent data indicate that RFs associated with seropositive RA are derived from terminally differentiated CD20-, CD38+ plasma cells (PCs) present in synovial fluids of the inflamed joints. These cells were shown to secrete RFs actively and are thought to originate from germinal centre (GC)-like structures present in the inflamed synovium. To obtain a representative image of the structural properties of IgM and IgG RFs associated with RA, phage antibody display libraries were constructed from CD38+ PCs isolated from the inflamed joints of RF-seropositive patients with RA. Subsequently, human IgG Fc-binding monoclonal phage antibodies were selected and analysed. The data suggest that RA-associated RFs are encoded by a diverse set of VL and a more restricted set of VH regions. VH gene family usage of PC derived IgM- and IgG-RFs was found to be restricted to the VH1 and 3 gene families, with a preference for VH3, and many different VL genes were shown to contribute to RF specificity. Clonally related VH as well as VL sequences were identified, based on the presence of identical CDR3 regions and shared somatic mutations. In this B cell selection process base-pair substitutions as well as deletions of triplets in CDR regions, leaving the transcripts in frame, were involved. Together, these data provide further evidence for an Ag-driven immune response in the terminal differentiation into RF-producing PCs in patients with RA, including expansion of clonally related B cells, selection and isotype switching, all hallmarks of a GC reaction. PMID- 12562402 TI - T(H)1/T(H)2 cytokine profile, metalloprotease-9 activity and hormonal status in pregnant rheumatoid arthritis and systemic lupus erythematosus patients. AB - During the course of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), several immune and neuroendocrine changes associated with pregnancy may exert positive (amelioration) or negative (exacerbation) effects on the clinical outcome. In order to shed light on the mechanisms underlying these responses, we performed a prospective longitudinal study in RA and SLE pregnant women, including healthy pregnant women as a control group. Cytokine messenger RNA (mRNA) expression assessed by quantitative competitive polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC), cytokine levels and lymphocyte proliferation responses (LPR) following phytohaemagglutinin (PHA) stimulation of PBMC, plasma metalloprotease-9 activity (MMP-9) and hormonal status during pregnancy were determined. TNFa was the most abundant cytokine mRNA expressed in PBMC in all groups studied (healthy pregnant women, RA and SLE pregnant patients). However, a general TH2 response reflected by high IL-10 levels was found in RA, as well as SLE, patients. A significant change in IFN gamma was observed in RA patients but only during the first trimester of pregnancy. This compared with a major TH1 response in healthy pregnant women. Interestingly, our study showed a homogeneous hormonal pattern in RA and SLE patients. Although decreased cortisol levels were observed in all patients studied, this is possibly related to the remission of disease activity status brought about by steroid treatment before and during pregnancy. In summary, we suggest that complex immune and hormonal networks are involved in pregnancy and that rheumatic diseases are very dynamic immune processes that cannot be described with a clear-cut cytokine profile. Furthermore, the observations in this study may reflect treatment-related immune effects more than those associated with disease. PMID- 12562404 TI - Novel airway devices: spoilt for choice? PMID- 12562403 TI - Modulation of autoreactive responses of peripheral blood lymphocytes of patients with systemic lupus erythematosus by peptides based on human and murine anti-DNA autoantibodies. AB - Two peptides, based on the sequences of the complementarity-determining regions (CDR) 1 and 3 of a pathogenic murine monoclonal anti-DNA autoatibody that bears the 16/6 idiotype (Id), were shown to either prevent or treat an already established systemic lupus erythematosus (SLE) in two murine models of lupus. Two additional peptides based on the human monoclonal anti-DNA, 16/6 Id were synthesized. This study was undertaken in order to investigate the ability of the CDR-based peptides to immunomodulate SLE-associated responses of peripheral blood lymphocytes (PBL) of SLE patients. PBL of 24 of the 62 SLE patients tested proliferated in vitro following stimulation with the human 16/6 Id. Peptides based on the CDRs of both the human and murine anti-DNA autoantibodies inhibited efficiently and specifically the 16/6 Id-induced proliferation and IL-2 production. The latter inhibitions correlated with an up-regulated production (by 2.5-3.5-fold) of the immunosuppressive cytokine, TGF-beta. Overall, the results of our study demonstrate that the CDR-based peptides are capable of down regulating in vitro autoreactive T cell responses of PBL of SLE patients. Thus, these peptides are potential candidates for a novel specific treatment of SLE patients. PMID- 12562405 TI - Alveolar recruitment strategy improves arterial oxygenation after cardiopulmonary bypass. AB - Atelectasis occurs during general anaesthesia. This is partly responsible for the impairment of gas exchange that occurs peri-operatively. During cardiopulmonary bypass, this atelectasis is exacerbated by the physical collapse of the lungs. As a result, poor arterial oxygenation is often seen postoperatively. We tested the effect of an 'alveolar recruitment strategy' on arterial oxygenation in a prospective randomised study of 78 patients undergoing cardiopulmonary bypass. Patients were divided equally into three groups of 26. Group 'no PEEP' received a standard post bypass manual lung inflation, and no positive end-expiratory pressure was applied until arrival at intensive care unit. Group '5 PEEP' received a standard post bypass manual inflation, and then 5 cmH2O of positive end-expiratory pressure was applied and maintained until extubation on intensive care. The third group, 'recruitment group', received a pressure-controlled stepwise increase in positive end-expiratory pressure up to 15 cmH2O and tidal volumes of up to 18 ml x kg(-1) until a peak inspiratory pressure of 40 cmH2O was reached. This was maintained for 10 cycles; the positive end-expiratory pressure of 5 cmH2O was maintained until extubation on intensive care. There was a significantly better oxygenation in the recruitment group at 30 min and 1 h post bypass when compared with the no PEEP and 5 PEEP groups. There was no significant difference in any of the groups beyond 1 h. Application of 5 cmH2O positive end expiratory pressure alone had no significant effect on oxygenation. No complications due to the alveolar recruitment manoeuvre occurred. We conclude that the application of an alveolar recruitment strategy improves arterial oxygenation after cardiopulmonary bypass surgery. PMID- 12562406 TI - Pre-operative screening: criteria for referring to anaesthetists. AB - Nurses assess patients pre-operatively using screening questionnaires and locally developed protocols. Our objectives were to determine which questions might identify patients who should be seen by an anaesthetist before the day of surgery. A review of the literature and a preliminary questionnaire to establish questions to be tested was followed by a modified, two-round Delphi questionnaire to determine the level of agreement by anaesthetists. There was agreement for referring patients who gave a positive response to questions that query: restricted exercise tolerance; previous anaesthetic problems; family history of anaesthetic problem; pathology affecting neck movement; angina; arrhythmia; heart failure; asthma; epilepsy; insulin-dependent diabetes mellitus; liver disease and unspecified kidney disease. There was equivocal agreement on questions that report a myocardial infarction over one year ago, cerebrovascular accident, non insulin-dependent diabetes mellitus and thyroid disease. Nurses should use these criteria during pre-operative assessment to decide the timing of evaluation by an anaesthetist. PMID- 12562407 TI - Multivariate analysis of factors associated with umbilical arterial pH and standard base excess after Caesarean section under spinal anaesthesia. AB - We have investigated the factors predicting umbilical arterial pH (UA pH) and standard base excess (UA BE) in 337 consecutive elective Caesarean sections performed under spinal anaesthesia. Multiple linear regression analysis was performed with UA pH and UA BE as the dependent factors. We found that the significant factors predicting UA pH were: use of ephedrine, uterine incision-to delivery time, maximum decrease in systolic arterial pressure and the interaction between ephedrine use and duration of hypotension (adjusted R2 = 0.39, F15,321 = 15.4, p < 0.0001). The significant factors predicting UA BE were: use of ephedrine and the interaction between ephedrine use and duration of hypotension (adjusted R2 = 0.52, F15,321 = 25.0, p < 0.0001). We conclude that, in order to minimise the risk of fetal acidosis, ephedrine should not be used before delivery, uterine incision-to-delivery time should be as short as possible, and alpha-agonists such as metaraminol or phenylephrine should be used to minimise both the magnitude and duration of hypotension. PMID- 12562409 TI - Laboratory and clinical comparisons of the Streamlined Liner of the Pharynx Airway (SLIPA) with the laryngeal mask airway. AB - The Streamlined Liner of the Pharynx Airway (SLIPA) is a new inexpensive disposable supraglottic airway designed to seal without the use of an inflatable cuff. It comprises a hollow blow-moulded soft plastic airway shaped to form a seal in the pharynx. Being hollow, liquid entrapment is possible and this may provide effective protection against aspiration. A model silicone rubber pharynx with an 'oesophageal' tube for injecting volumes of regurgitant liquid was designed to evaluate the SLIPA and the standard and ProSeal laryngeal mask airways during positive-pressure ventilation. A linear relationship between the volume 'regurgitated' and the volume 'aspirated' was found with the laryngeal mask airway and the ProSeal laryngeal mask airway with the drainage tube clamped. Both the ProSeal laryngeal mask airway with an open drainage tube and the SLIPA, but not the standard laryngeal mask airway, provided effective protection against 'aspiration' during positive-pressure ventilation using the model. In a clinical study, 120 patients were randomly allocated to receive controlled ventilation of the lungs via the standard laryngeal mask airway or the SLIPA. Both devices were equally easy to insert and satisfactory for airway management. PMID- 12562408 TI - Effect of intra-operative magnesium sulphate on pain relief and patient comfort after major lumbar orthopaedic surgery. AB - The effects of intra-operative magnesium sulphate on pain relief after major lumbar surgery were investigated in 24 patients. Patients were randomly allocated to receive either an infusion of 50 mg x kg(-1) magnesium sulphate or an equivalent volume of saline at induction of anaesthesia. Anaesthesia was induced with propofol and remifentanil. Tracheal intubation was facilitated using rocuronium. Maintenance was achieved with remifentanil and sevoflurane in nitrous oxide/ oxygen. Intra-operative monitoring included standard equipment and neuromuscular transmission. During surgery, neuromuscular block recovery was longer in the magnesium group. Postoperative opioid consumption and pain scores were lower in the magnesium group. The first night's sleep and the global satisfaction scores were better in the magnesium group. The results of the study support magnesium sulphate as a useful adjuvant for postoperative analgesia after major lumbar surgery. PMID- 12562410 TI - Assessment of the Ventrix parenchymal intracranial pressure monitoring probe (NL950-P) and Monitor (NL950-100) in a 3 Tesla magnetic resonance scanner. AB - Magnetic resonance (MR) imaging and spectroscopy provide important information in patients with acute head injury. However, optimal patient management requires intracranial pressure (ICP) monitoring. There are few reports on the use of ICP sensors in an MR environment. We tested the Ventrix parenchymal intracranial pressure monitoring probe and monitor (Integra Neurosciences, USA), modified by the use of a fibre-optic extension cable, within a 3 Tesla MR system. The device performed well in the MR environment, but one element within the fibre-optic extension was significantly ferromagnetic. The ICP probe produced a small susceptibility artefact on spin echo images, and a larger artefact on gradient echo images. The MR safety of the integrated system is probably acceptable, but could be easily improved with minor modifications. Although the system is MR compatible and produces generally acceptable imaging even at 3 Tesla, there is significant degradation of image quality during gradient echo sequences. PMID- 12562411 TI - Mainstream vs. sidestream capnometry for prediction of arterial carbon dioxide tension during supine craniotomy. AB - We compared the performance of mainstream capnometry as a measure of arterial carbon dioxide tension (Paco2) with sidestream recordings in adult neurosurgical patients undergoing supine craniotomy. Two hundred and forty patients were randomly assigned so that the end-tidal carbon dioxide tension (PEco2) was measured using either a mainstream or sidestream infrared capnometer. All patients received propofol anaesthesia and ventilation was adjusted according to clinical requirement. Arterial blood gas analyses were performed after induction, prior to dural incision, during surgery and before wound closure. Simultaneous haemodynamic and ventilatory parameters were also recorded. For 1007 paired measurements of PEco2 and Paco2 (mainstream, n = 503; sidestream, n = 504), the mean (SD) mainstream arterial to end-tidal carbon dioxide tension difference, 0.64 (0.16) kPa, was smaller than the corresponding sidestream values, 0.99 (0.40) kPa (p < 0.001). The limits of agreement for the mainstream analyser, 0.32 0.96 kPa, were also narrower than the sidestream recordings, 0.19-1.79 kPa (p < 0.001). In both capnometers, the arterial to end-tidal difference in carbon dioxide tension did not change with time. However, there was greater within patient variation in the sidestream group. Our study showed that mainstream PEco2 provided a more accurate estimation of Paco2 than sidestream measurement. PMID- 12562412 TI - Calibration of three capnographs for use with helium and oxygen gas mixtures. AB - Capnography is considered essential in the management of mechanically-ventilated patients. Helium, as an adjunct to mechanical ventilation, is the subject of renewed interest and used increasingly. However, helium affects the performance of infrared capnometry. We constructed a simple device to generate variable mixtures of helium, oxygen and carbon dioxide within the normal physiological range, and tested the performance of two side-stream and one in-line capnographs. We found that addition of helium to the gas mixture caused all three capnographs to underestimate the concentration of carbon dioxide. The underestimation increased as the proportion of helium increased. The maximum underestimation (30%) occurred in a 79:21 helium/oxygen mixture. PMID- 12562413 TI - Peri-operative optimisation. PMID- 12562414 TI - Making herbal medicines safe. PMID- 12562415 TI - The role of evidence-based methods in scientific study. PMID- 12562416 TI - Unfinished business. PMID- 12562417 TI - Emergency physicians: additional providers of emergency anaesthesia? PMID- 12562418 TI - Airway management training for all. PMID- 12562419 TI - A 'foreign body' in the circuit filter. PMID- 12562420 TI - Unexpected difficult intubation with Portex tracheal tube introducer. PMID- 12562421 TI - Post intubation airway obstruction in thyroid surgery. PMID- 12562422 TI - Treatment of laryngospasm. PMID- 12562423 TI - Another use for capnography. PMID- 12562424 TI - Improved submental intubation. PMID- 12562425 TI - Finding a use for the lumen in the Portex Tracheal Tube Guide. PMID- 12562426 TI - Oxygen delivery failure. PMID- 12562427 TI - An evaluation of the PAxpress pharyngeal airway. PMID- 12562428 TI - Bougie trauma--what trauma? PMID- 12562429 TI - Air-oxygen flowmeter confusion. PMID- 12562430 TI - Bronchospasm and difficult intubation. PMID- 12562431 TI - Anaphylaxis to rocuronium. PMID- 12562432 TI - Remifentanil for phaeochromocytoma resection. PMID- 12562433 TI - Cricoid ring thing. PMID- 12562434 TI - Sedation using remifentanil. PMID- 12562435 TI - Todd's paralysis following sedation for sleep nasendoscopy. PMID- 12562436 TI - Local clinical research and changes in local clinical practice. PMID- 12562437 TI - A vaginal ultrasound probe is better than nothing. PMID- 12562438 TI - Bretylium in the treatment of complex regional pain syndrome: uncommon side effect of a common drug. PMID- 12562439 TI - Donepezil and succinylcholine. PMID- 12562440 TI - Ipsilateral arm swelling--a rare complication of jugular venous catheterisation. PMID- 12562441 TI - Blood bath. PMID- 12562442 TI - Review article: Helicobacter pylori and reflux disease. AB - The falling prevalence of Helicobacter pylori infection and related diseases (peptic ulcer disease, gastric cancer) in developed countries has been paralleled by an increased recognition of gastro-oesophageal reflux and its complications. These epidemiological data do not support a role for H. pylori in the pathogenesis of reflux disease, but suggest a negative association with the increasing incidence of oesophageal diseases. This has led some investigators to propose a 'protective' role of H. pylori infection against the development of oesophageal diseases. In these patients, pre-existing lower oesophageal sphincter dysfunction, susceptibility to reflux, unmasking of latent reflux and the patterns and severity of gastritis are probably important factors contributing to the development of oesophageal diseases. The most likely mechanism by which H. pylori infection may protect against reflux is by decreasing the potency of the gastric refluxate in patients with corpus-predominant gastritis. The prevalence of H. pylori infection in patients with reflux disease is probably no greater than that in those without reflux, and there are conflicting data indicating that reflux symptoms or erosive oesophagitis develop after H. pylori eradication. It is also unclear whether H. pylori augments the antisecretory effects of proton pump inhibitors or accelerates the development of atrophic gastritis. PMID- 12562443 TI - Review article: nutrition and adult inflammatory bowel disease. AB - Major advances in the understanding of the aetio-pathogenesis and genetics of inflammatory bowel disease have been accompanied by an escalation in the sophistication of immunomodulatory inflammatory bowel disease therapeutics. However, the basic 'triple' therapy (5-aminosalicylates, corticosteroids, azathioprine) and nutrition have maintained their central role in the management of patients with inflammatory bowel disease over recent decades. This review provides an overview of the supportive and therapeutic perspectives of nutrition in adult inflammatory bowel disease. The objective of supportive nutrition is to correct malnutrition in terms of calorie intake or specific macro- or micronutrients. Of particular clinical relevance is deficiency in calcium, vitamin D, folate, vitamin B12 and zinc. There is justifiably a growing sense of unease amongst clinicians and patients with regard to the long-term use of corticosteroids in inflammatory bowel disease. This, rather than arguments about efficacy, should be the catalyst for revisiting the use of enteral nutrition as primary treatment in Crohn's disease. Treatment failure is usually related to a failure to comply with enteral nutrition. Potential factors that militate against successful completion of enteral nutrition are feed palatability, inability to stay on a solid-free diet for weeks, social inconvenience and transient feed related adverse reactions. Actions that can be taken to improve treatment outcome include the provision of good support from dietitians and clinicians for the duration of treatment and the subsequent 'weaning' period. There is evidence to support a gradual return to a normal diet through exclusion-re-introduction or other dietary regimen following the completion of enteral nutrition to increase remission rates. We also review the evidence for emerging therapies, such as glutamine, growth factors and short-chain fatty acids. The future may see the evolution of enteral nutrition into an important therapeutic strategy, and the design of a 'Crohn's disease-specific formulation' that is individually tailored, acceptable to patients, cost-effective, free from adverse side-effects and combines enteral nutrition with novel pre- and pro-biotics and other factors. PMID- 12562444 TI - Review article: NSAIDs, gastroprotection and cyclo-oxygenase-II-selective inhibitors. AB - In patients at high risk of NSAID-associated serious upper gastrointestinal complications, gastroprotection with misoprostol or a proton pump inhibitor should be considered. Only misoprostol, 800 micro g/day, has been shown to reduce serious upper gastrointestinal complications in a large clinical outcome trial. The benefit of Helicobacter pylori eradication in reducing NSAID-associated gastrointestinal toxicity is controversial, and routine testing for and eradication of H. pylori in NSAID users are not currently advised. The gastrointestinal safety of rofecoxib and celecoxib has been assessed in large clinical outcome trials which, on first analysis, show benefits over non selective NSAIDs in the incidence of serious upper gastrointestinal complications. However, longer term gastrointestinal data from the celecoxib study (CLASS) and cardiovascular adverse event data from the rofecoxib study (VIGOR) have questioned the risk-benefit profile of these new drugs and, until they are better understood, it seems sensible not to use them routinely in large numbers of individuals. The gastrointestinal safety of meloxicam and etodolac has not been adequately assessed in such trials. Therefore, evidence for their use instead of non-selective NSAIDs, or instead of celecoxib or rofecoxib, is not robust. PMID- 12562445 TI - Esomeprazole 20 mg and lansoprazole 15 mg in maintaining healed reflux oesophagitis: Metropole study results. AB - AIM: To compare the efficacy of esomeprazole, 20 mg once daily, vs. lansoprazole, 15 mg once daily, for the maintenance treatment of patients with healed reflux oesophagitis. METHODS: During the initial open healing phase, 1391 patients with endoscopically verified reflux oesophagitis and a history of heartburn, with or without acid regurgitation, received esomeprazole 40 mg for 4-8 weeks. Patients who were healed (identified by endoscopy at 4 or 8 weeks) and symptom free were then randomized to receive 6 months of treatment with esomeprazole, 20 mg once daily, or lansoprazole, 15 mg once daily. RESULTS: Esomeprazole, 20 mg once daily, maintained a significantly higher proportion of patients in remission than lansoprazole, 15 mg once daily, over 6 months [83% (95% CI, 80-86%) of esomeprazole recipients compared with 74% (95% CI, 70-78%) of lansoprazole recipients; P < 0.0001; life table estimates]. When data were analysed according to baseline Los Angeles grade classification, esomeprazole, 20 mg once daily, achieved consistently higher remission rates across all grades of disease severity, whereas the efficacy of lansoprazole decreased to a greater extent with increasing severity of reflux oesophagitis. CONCLUSION: Esomeprazole, 20 mg once daily, is more effective than lansoprazole, 15 mg once daily, in maintaining remission in patients with healed reflux oesophagitis. PMID- 12562446 TI - A randomized, double-blind trial of the efficacy and safety of 10 or 20 mg rabeprazole compared with 20 mg omeprazole in the maintenance of gastro oesophageal reflux disease over 5 years. AB - BACKGROUND: Gastro-oesophageal reflux disease has a chronic course, and often requires long-term treatment. Proton pump inhibitors are the treatment of choice for both acute and maintenance treatment, but little is known from randomized controlled trials of their effects beyond 1 year. AIM: To compare the efficacy and safety of two doses of rabeprazole with 20 mg omeprazole in the maintenance treatment of erosive gastro-oesophageal reflux disease over 5 years. METHODS: Two hundred and forty-three patients who had previously responded to acute treatment for erosive gastro-oesophageal reflux disease were prospectively randomized to receive 5 years of treatment with rabeprazole (10 or 20 mg daily) or omeprazole (20 mg daily). The primary outcome measure was endoscopically confirmed relapse of erosive gastro-oesophageal reflux disease. RESULTS: One hundred and twenty three patients (51%) completed all 5 years of the study, with similar completion rates in the three groups. Relapses occurred in nine of 78 (11.5%), eight of 82 (9.8%) and 11 of 83 (13.3%) patients in the rabeprazole 20 mg, rabeprazole 10 mg and omeprazole 20 mg groups, respectively. Gastric biopsy showed no evidence of any harmful effects. All treatments were well tolerated. CONCLUSIONS: Rabeprazole 10 mg, rabeprazole 20 mg and omeprazole 20 mg all had similar efficacy in the maintenance treatment of gastro-oesophageal reflux disease. All three were safe and well tolerated during 5 years of treatment. PMID- 12562447 TI - Circadian pattern of intragastric acidity in patients with non-erosive reflux disease (NERD). AB - BACKGROUND: Most patients with gastro-oesophageal reflux disease have non-erosive reflux disease. Proton pump inhibitors are less effective than expected in these patients, but no previous study has measured their 24-h gastric pH values. AIMS: To evaluate whether there are differences in 24-h intragastric acidity between reflux patients with and without oesophagitis and controls. The influence of Helicobacter pylori on the gastric pH of reflux patients was also assessed. METHODS: Sixty-three consecutive patients with gastro-oesophageal reflux disease symptoms who agreed to undergo endoscopy and 24-h pH-metry were recruited. Twenty five (39%) had erosive oesophagitis and 38 (61%) did not. H. pylori was diagnosed by CLO test, histology and 13C-urea breath test. Gastric pH was also measured in 30 controls without digestive symptoms. RESULTS: H. pylori was found in seven of the 25 (28%) patients with oesophagitis and 14 of the 38 (37%) patients with non erosive reflux disease. Oesophageal pH-metry was abnormal in 21 of the 25 (84%) patients with oesophagitis and in 32 of the 38 (84%) patients with non-erosive reflux disease. The median gastric pH did not differ between patients with and without oesophagitis or between them and controls during the 24 h (P = 0.8) and other time intervals (P = 0.2-0.4). The gastric pH did not differ between infected and non-infected patients with oesophagitis (P = 0.2-0.4) or non-erosive reflux disease (P = 0.3-0.8). CONCLUSIONS: The circadian pattern of intragastric acidity does not differ between patients with non-erosive reflux disease and oesophagitis. Moreover, the study confirms that H. pylori infection does not affect the gastric pH in either group of reflux patients. PMID- 12562448 TI - Comparative pharmacokinetics and safety of lansoprazole oral capsules and orally disintegrating tablets in healthy subjects. AB - BACKGROUND: Many individuals with acid-related gastrointestinal disorders have difficulty in swallowing oral agents. AIM: To compare the bio-availability of a single dose of lansoprazole orally disintegrating tablet with that of an intact capsule. METHODS: One hundred and twenty healthy subjects participated in two prospective, Phase I, open-label, two-period cross-over studies to receive lansoprazole, 15 mg or 30 mg. Within each study, subjects were randomized into two parallel cohorts consisting of 30 subjects per regimen, dispensed in opposing sequence over two periods separated by a 7-day washout period. Blood samples were collected on day 1 of both periods to determine the pharmacokinetic parameters. RESULTS: Tmax occurred at 1.8 and 2.0 h with the 15-mg and 30-mg tablets, respectively. Dose proportional increases in Cmax, AUCt and AUC infinity were observed in the 15-mg and 30-mg groups. The terminal elimination half-lives (t1/2) were identical in both dose groups (1.18 h). Lansoprazole administered as the orally disintegrating tablet was bio-equivalent to the intact capsule formulation with respect to Cmax, AUCt and AUC infinity. CONCLUSIONS: Lansoprazole orally disintegrating tablets, 15 mg and 30 mg, are bio-equivalent to the respective dose administered as the intact capsule. This novel dosage formulation represents an option for patients who have difficulty in swallowing oral agents. PMID- 12562449 TI - Symptomatic response to lansoprazole predicts abnormal acid reflux in endoscopy negative patients with non-cardiac chest pain. AB - AIM: To determine whether symptomatic response to lansoprazole predicts abnormal acid reflux in endoscopy-negative patients with non-cardiac chest pain. METHODS: Patients who complained of chest pain, but had normal coronary angiography, were asked to undergo upper endoscopy. Those without gastric and oesophageal lesions were recruited for 24-h ambulatory oesophageal pH monitoring, and were randomly given lansoprazole 30 mg or placebo, both daily for 4 weeks. Chest pain symptoms were recorded before and 1 month after treatment on a locally validated questionnaire. The symptom score was calculated by multiplying the severity and frequency of the symptom, and symptom improvement was defined as > 50% reduction in symptom score. RESULTS: Overall, 68 patients, 36 on lansoprazole and 32 on placebo, completed the trial. The symptom score was reduced significantly in both groups (P < 0.001). In the lansoprazole group, more patients with than without abnormal reflux showed symptom improvement (92% vs. 33%; odds ratio = 22; 95% confidence interval, 2.3-201.8; chi2 = 10.9; P = 0.001), giving a sensitivity, specificity, positive predictive value, negative predictive value and accuracy of 92%, 67%, 58%, 94% and 75%, respectively. In the placebo group, the rates of symptom improvement were similar between those with and without abnormal reflux (33% vs. 35%, P = N.S.). CONCLUSIONS: Treatment with lansoprazole is a useful test in diagnosing endoscopy-negative gastro-oesophageal reflux disease in Chinese patients with non-cardiac chest pain. PMID- 12562450 TI - Cyclo-oxygenase-2 expression in Barrett's oesophageal carcinogenesis: an immunohistochemical study. AB - BACKGROUND: The incidence of Barrett's oesophageal adenocarcinoma is increasing more rapidly than any other malignancy in industrialized countries. Cyclo oxygenase-2 appears to play an important role in gastrointestinal carcinogenesis. Previous studies on cyclo-oxygenase-2 expression in Barrett's oesophageal carcinogenesis have utilized tissue samples obtained from different patients. We sought a definitive comparison of cyclo-oxygenase-2 expression in the sequence of Barrett's metaplasia-dysplasia-adenocarcinoma within the same patients. METHODS: Paraffin-embedded oesophago-gastrectomy specimens from 20 patients, containing successive stages of Barrett's metaplasia, high-grade dysplasia and adenocarcinoma, were analysed for cyclo-oxygenase-2 expression by immunohistochemistry. RESULTS: Cyclo-oxygenase-2 was constitutively expressed in the basal layers of cells in the adjacent normal squamous oesophageal epithelium, but a higher cyclo-oxygenase-2 expression was observed in Barrett's metaplasia. A further increase in cyclo-oxygenase-2 expression was detected in high-grade dysplasia, but cyclo-oxygenase-2 was decreased in adenocarcinoma tissue, regardless of its stage or level of differentiation. CONCLUSIONS: Cyclo-oxygenase 2 expression is progressively increased when squamous oesophageal epithelium develops into Barrett's metaplastic epithelium and then into high-grade dysplasia, but appears to decrease when adenocarcinoma develops. These findings may be significant for an effective chemo-prevention strategy with selective cyclo-oxygenase-2 inhibitors. PMID- 12562451 TI - Response of fistulating Crohn's disease to infliximab treatment assessed by magnetic resonance imaging. AB - AIMS: To assess fistula track healing after infliximab treatment using magnetic resonance imaging. METHODS: Magnetic resonance imaging and clinical evaluation were performed before and after three infliximab infusions given over a 6-week period. Magnetic resonance images were evaluated for abscesses and fistula tracks. Paired magnetic resonance image examinations were rated 'better', 'unchanged' or 'worse'. Magnetic resonance imaging and clinical outcomes were then compared. RESULTS: Of the 12 referred patients, pre-treatment magnetic resonance imaging detected abscesses in three (two not treated). Of the 10 treated patients, seven had peri-anal fistulas, two of whom also had recto vaginal fistulas, and three had abdominal wall entero-cutaneous fistulas. After infliximab, four were in remission, one had a response and five were non responders. One developed a peri-anal abscess. Magnetic resonance imaging improved in six, was unchanged in two and was worse in two. In four of the six with improvement in magnetic resonance imaging, the fistula track resolved, but two of these had clinically persistent entero-cutaneous fistulas. The clinical outcome and magnetic resonance imaging correlated in seven of the 10 patients; in three (two entero-cutaneous and one peri-anal), there was discordance. CONCLUSIONS: Magnetic resonance imaging identifies clinically silent sepsis. Fistulas may persist despite clinical remission. Clinical response to infliximab and clinical correlation with magnetic resonance imaging were poor in patients with abdominal entero-cutaneous fistulas. PMID- 12562452 TI - Gastrointestinal transit and 5-ASA release from a new mesalazine extended-release formulation. AB - BACKGROUND: Mesalazine (5-aminosalicylic acid, 5-ASA)-containing formulations represent a cornerstone in the treatment of inflammatory bowel diseases. Recently, a new formulation has been developed to provide selective and more homogeneous release of 5-ASA compared to traditional systems. METHODS: In a first study, gastrointestinal transit was followed by gamma-scintigraphy after single dose application of tablets containing 1200 mg mesalazine to 12 healthy male volunteers. 5-ASA release was verified by the assessment of plasma pharmacokinetics. In a second, 7-day, multiple-dose study, the steady state plasma pharmacokinetics, urinary excretion and safety profile were characterized after twice-daily tablet administration to 12 healthy volunteers. RESULTS: Tablet erosion started after 6.9 +/- 1.1 h in the ascending or transverse colon. Radioactivity spread homogeneously throughout the colon, indicating the sustained release of active 5-ASA. Plasma kinetics indicated an earlier initial absorption of 5-ASA, i.e. during transit of the small intestine and ileum. Mean Cmax values (350.6 +/- 322.6 ng/mL) were observed during location in the ileo-caecal region. The mean relative absorption of 5-ASA was 19.9 +/- 18.2% in the small intestine and ileum and 80.1 +/- 18.2% in the colon. CONCLUSIONS: The administration of the new mesalazine formulation was well tolerated, and 5-ASA was continuously released along the whole colon, a favourable prerequisite for the therapy of distally located inflammatory bowel disease. PMID- 12562453 TI - Pharmacokinetics of budesonide controlled-release capsules when taken with omeprazole. AB - AIM: To investigate whether omeprazole affects the pharmacokinetics and systemic effects of budesonide controlled-release capsules when the two medications are taken together. METHODS: Thirteen healthy volunteers were enrolled into a randomized, double-blind, placebo-controlled, cross-over study. Participants received omeprazole, 20 mg/day, or placebo every morning for 5 days, with three 3 mg budesonide controlled-release capsules being given with omeprazole or placebo on day 5. After a 12-day washout period, participants were switched from omeprazole to placebo, or vice versa, and the trial was repeated. Blood samples for pharmacokinetic evaluation and urine samples for cortisol assessments were collected before and after the budesonide doses. RESULTS: No statistically significant differences were seen between omeprazole and placebo treatment with regard to any of the parameters analysed, including the maximum budesonide plasma concentration, time to concentration maximum, area under the concentration-time curve, mean residence time and urinary excretion of cortisol. Very few adverse events were reported during the trial, and the majority were of mild to moderate severity. CONCLUSION: Omeprazole treatment does not affect the pharmacokinetics or systemic effects of budesonide controlled-release capsules when the two medications are taken simultaneously. PMID- 12562454 TI - Patients with refractory Crohn's disease or ulcerative colitis respond to dehydroepiandrosterone: a pilot study. AB - BACKGROUND: Dehydroepiandrosterone is a steroid hormone used as an 'over-the counter' drug in the USA. Treatment with dehydroepiandrosterone was effective in randomized controlled trials in patients with systemic lupus erythematosus. Dehydroepiandrosterone sulphate concentrations are decreased in patients with inflammatory bowel disease. Dehydroepiandrosterone inhibits nuclear factor-kappaB and the secretion of interleukin-6 and interleukin-12 via the peroxisome proliferator-activated receptor alpha. AIM: A phase II pilot trial was started to evaluate the effect of dehydroepiandrosterone in active inflammatory bowel disease. METHODS: Twenty patients with chronic active inflammatory bowel disease [seven Crohn's disease (Crohn's disease activity index, 242 +/- 51; mean +/- s.d.); 13 ulcerative colitis (clinical activity index, 7.8 +/- 2.1)] took 200 mg dehydroepiandrosterone per day orally for 56 days. RESULTS: Six of the seven patients with Crohn's disease and eight of the 13 patients with ulcerative colitis responded to treatment, with a decrease in the Crohn's disease activity index of > 70 points and a decrease in the clinical activity index of > 4 points, respectively. Six Crohn's disease patients and six ulcerative colitis patients went into remission (Crohn's disease activity index < 150; clinical activity index 0 to < 4 months, 4-6 months and> 6 months. Associations between breast feeding duration and PPVT-R at 6 years and Block Design at 8 years were estimated before and after adjustment for gender, gestational age, maternal age, maternal education, parental smoking and the presence of older siblings. The early cessation of full breast feeding was associated with reduced verbal IQ and the performance subtest. In unadjusted analysis, mean standardised PPVT-R scores were 6.44 points greater (P < 0.0001) in children fully breast fed for> 6 months compared with those never breast fed. After adjustment, mean PPVT-R scores were 3.56 points higher in children fully breast fed for> 6 months compared with those children never breast fed (P = 0.003). Similarly, Block Design scores were higher in those fully breast fed for> 6 months compared with those never breast fed in unadjusted (P = 0.001) but not adjusted analyses (P = 0.223). Interactions between maternal education (four levels) and breast feeding demonstrated a positive association of maternal education on verbal IQ (F = 2.64; P = 0.005) in children breast fed for longer but not on performance (F = 0.74; P = 0.67). The early introduction of milk other than breast milk was associated with reduced verbal IQ after adjustment for social and perinatal confounders. Although these effects were interacting with maternal education, they may act through undefined mechanisms in human milk. PMID- 12562476 TI - Maternal factors associated with the duration of breast feeding in Jeddah, Saudi Arabia. AB - Recently, there has been increasing concern about the decline in breast-feeding pattern in developing countries. The objectives of this study were to document the recent breast-feeding trends in Jeddah during the first year of an infant's life and identify the probable maternal risk factors implicated in breast-feeding cessation. Data were collected from six randomly selected primary health care centres in Jeddah City. All married women with an infant 0.05). The time to recovery to an Aldrete score of 10 was significantly shorter in group B than group BM (P < 0.05). Although not significant, it was also shorter in group B than group BF. There was no difference in additional analgesic requirements between the groups in the first 24 h. Sedation score was higher in the midazolam group at 60 and 90 min postoperatively than the other groups. CONCLUSIONS: Caudal block with 0.75 ml.kg( 1) 0.25% bupivacaine and 50 microg.kg(-1) midazolam or 1 microg.kg(-1) fentanyl provides no further analgesic advantages to bupivacaine alone when administered immediately after induction of anaesthesia in children undergoing unilateral inguinal herniorrhaphy. PMID- 12562486 TI - Parents' perceptions and use of analgesics at home after children's day surgery. AB - BACKGROUND: Children are found to suffer from unnecessarily severe postoperative pain following day surgery. Reasons for parents' insufficient use of analgesics may be based on misleading perceptions of children's analgesics. The purpose of this study was to describe parents' perceptions and use of analgesics for children after discharge at home. METHODS: In this survey, 840 questionnaires were given to parents, and answers from 201 mothers and 114 fathers whose children, aged 1-6 years, had undergone day surgery in 10 Finnish hospitals are presented. RESULTS: One-third (36%) of the children were assessed as having moderate or severe postoperative pain after discharge. More than three-quarters of the parents had given analgesics to the child. Analgesics were given mostly to children who were assessed to have pain and found to have several types of pain behaviour. Most of the parents had accurate perceptions of children's analgesics, but some of them had misleading perceptions of the nature and adverse effects of children's analgesics, which were related to giving analgesics to the child. Fathers, more often than mothers, seem to have such misleading perceptions. CONCLUSIONS: Parents tended to give analgesics to children who actually needed pain alleviation. Parents' perceptions of children's analgesics were mostly accurate. However, misleading perceptions of children's analgesics also exist among the parents. These misleading perceptions appear to decrease parents' use of children's postoperative pain medication. PMID- 12562487 TI - A comparison of five techniques for detecting cardiac activity in infants. AB - BACKGROUND: The new guidelines for cardiopulmonary resuscitation recommend that laypersons should begin chest compressions without checking for a pulse because the pulse check has serious limitations in accuracy. We determined the efficacy of the most suitable method to search for cardiac activity in infants. METHODS: Twenty-eight nurses tried to detect infants' cardiac activity and determined their heart rates with five different techniques: palpation of brachial pulse, carotid pulse, femoral pulse, apical impulse and auscultation of apical impulse with the naked ear (direct auscultation technique). RESULTS: The mean time interval required to find the pulse within 30 s in the auscultation, the apical, the brachial, the carotid and the femoral were 2.4 +/- 1.2, 3.5 +/- 2.7, 4.0 +/- 2.7, 9.9 +/- 7.0 and 9.1 +/- 5.9 s, respectively. The required time was significantly shorter in the auscultation method than in the palpation of carotid and femoral pulses. The percentage and 95% confidence intervals (95% CI) of pulses identified within 10 s (= the number of the correct identified within 10 s/the number of all cases) in auscultation, apical, brachial, carotid and femoral palpations were 100.0% (95% CI 51.8, 100), 75.0% (95% CI 28.9, 89.3), 73.1% (95% CI 52.2, 88.4), 50.0% (95% CI 30.6, 69.4) and 42.9% (95% CI 24.5, 62.8), respectively. These values were greater in the auscultation method than in all the palpation methods. CONCLUSIONS: The direct auscultation technique was more rapid and accurate than any other techniques to determine cardiac activity without instruments. It is suggested that direct a auscultation technique is also superior to the palpation of brachial artery in cardiopulmonary resuscitation in infants. PMID- 12562488 TI - Sedation and neuromuscular blockade in paediatric intensive care: a review of current practice in the UK. AB - BACKGROUND: Our aim was to investigate the current practice of sedation and neuromuscular blockade in critically ill children in paediatric intensive care units (PICUs) in the UK. METHODS: A postal questionnaire was sent to all PICUs in the UK. RESULTS: The most commonly used sedative agents were midazolam in combination with morphine. Written clinical guidelines for the sedation of critically ill children were available in 45% of units. Sedation is formally assessed in 40% of units. Vecuronium is the most commonly used neuromuscular blocking agent. In the UK, 31% of critically ill children are likely to receive neuromuscular blocking agents. Depth of neuromuscular blockade is routinely assessed in 16% of patients. CONCLUSIONS: Relatively few units possess clinical guidelines for the sedation of critically ill children, and only a minority formally assess sedation levels. Where neuromuscular blocking agents are administered, sedation is frequently inadequately assessed and the depth of neuromuscular blockade is rarely estimated. PMID- 12562489 TI - Regional anaesthesia of the lower extremity in infants and children. PMID- 12562490 TI - Is ilioinguinal/iliohypogastric nerve block always totally safe in children? AB - We report a case of accidental puncture of the small bowel during an ilioinguinal/iliohypogastric nerve block procedure for hernia repair. The diagnosis was made a few days later during a laparoscopic exploration owing to the progressive onset of clinical and radiological intestinal obstruction. A large, obstructing subserosal haematoma was found without any apparent perforation of the mucosa, and the damaged loop was resected. Such a complication has already been reported once in the literature after use of a long bevel needle for the puncture. This case is the first reported using an atraumatic short bevel needle. We discuss the technical aspects of the procedure and underline the fact that regional anaesthesia in children is never totally risk free. PMID- 12562491 TI - Anaesthetic management in McKusick-Kaufman syndrome. AB - The cardinal features of McKusick-Kaufman syndrome (MKS) are polydactyly and hydrometrocolpos. Sometimes, this abdominopelvic mass may restrict pulmonary function and decrease the oxygenation of the newborn. We present a case of MKS and review the clinical features and appropriate anaesthetic management. A 45-day old girl, weighing 4450 g, born at term, presented to our Paediatric Surgery Department with abdominal distention, diarrhoea and vomiting, which started on the 15th day of life. On physical examination, peripheral cyanosis, tachycardia and tachypnoea were found. A prominent mass was palpated in the lower abdomen. There was vaginal atresia and a sixth digit was found on her left foot. There was a large cystic mass extending from the pelvis to the level of L1 displacing the diaphragm, and bilateral hydroureteronephrosis were seen on plain abdominal X ray, ultrasonography and computerized tomography. In the preanaesthetic assessment, the patient was evaluated for multiple congenital anomalies. Our management of anaesthesia, for a neonate with severe hydrometrocolpos compressing the diaphragm, consisted of gastric decompression and preoxygenation before tracheal intubation, routine and airway pressure monitoring, periodic analyses of blood gases and maintenance of anaesthesia with a low concentration of volatile agent, together with an opioid. PMID- 12562492 TI - Single-dose caudal anaesthesia for two infants undergoing diagnostic brain magnetic resonance imaging: high risk and nonhigh risk. AB - We present a case report of two infants given a bupivacaine caudal anaesthetic as a means of achieving sedation for a diagnostic brain magnetic resonance imaging (MRI). Patient 1 was born at 27 weeks of gestation and presented to our hospital at 39 weeks postconception with a history of bronchopulmonary dysplasia and apnoea/bradycardia spells. He was undergoing a brain MRI for the work up of persistent apnoea and craniofacial abnormalities. Patient 2 was a term infant at 42 weeks postconception undergoing a MRI for the work up of a seizure disorder. Both infants fell asleep shortly after placement of the caudal blocks and the studies were completed successfully without complications. Utilizing this technique, we avoided dealing with a potentially difficult airway (patient 1), exacerbating postoperative apnoea and the negative implications of intravenous sedation during a study where there is limited access to the patient's airway. PMID- 12562493 TI - Laryngotracheo-oesophageal cleft in an infant: airway management during bronchoscopy. AB - A case of laryngotracheo-oesophageal cleft scheduled for bronchoscopy and repair is described. The authors highlight the difficulties of bronchoscopy in such a patient and suggest simple manoeuvres to negotiate the rigid bronchoscope into the trachea. PMID- 12562494 TI - Rett syndrome: anaesthesia management. PMID- 12562496 TI - Counting the way to induction of anaesthesia. PMID- 12562497 TI - Laryngeal mask airway for ventilatory support over a 4-day period in a neonate with Pierre Robin sequence. PMID- 12562498 TI - Melatonin effects on bone: experimental facts and clinical perspectives. AB - Bone formation proceeds through a remodeling process that runs continuously, involving the resorption of old bone by osteoclasts, and the subsequent formation of new bone by osteoblasts. This is controlled by growth factors and cytokines produced in bone marrow microenvironment and by the action of systemic hormones, like parathyroid hormone, estradiol or growth hormone (GH). One candidate for hormonal modulation of osteoblast and osteoclast formation is melatonin. Because circulating melatonin declines with age, its possible involvement in post menopausal and senescence osteoporosis is considered. This review article discusses early studies on melatonin-bone relationships and recent data that suggest a direct effect of melatonin on bone. Melatonin could act as an autacoid in bone cells as it is present in high quantities in bone marrow, where precursors of bone cells are located. Melatonin dose-dependently augmented proteins that are incorporated into the bone matrix, like procollagen type I c peptide. Osteoprotegerin, an osteoblastic protein that inhibits the differentiation of osteoclasts is also augmented by melatonin in vitro. Another possible target cell for melatonin is the osteoclast, which degrades bone partly by generating free radicals. Melatonin through its free radical scavenger and antioxidant properties may impair osteoclast activity and bone resorption. At least in one study melatonin was both inhibitory to osteoclastic and osteoblastic cells. Therefore, the documented bone-protecting effect of melatonin in ovariectomized rats can depend in part on the free radical scavenging properties of melatonin. Additionally, melatonin may impair development of osteopenia associated with senescence by improving non-rapid eye movement sleep and restoring GH secretion. Whether melatonin can be used as a novel mode of therapy for augmenting bone mass in diseases deserves to be studied. PMID- 12562499 TI - The validity and feasibility of saliva melatonin assessment in the elderly. AB - Recent work in young and middle-aged subjects suggests that melatonin levels in saliva may represent a viable alternative to serum melatonin measurement. We hypothesized that it may be a valid measure of melatonin levels in older adults as well, but features unique to the elderly may limit its utility. To study this, subjects were admitted to an academic medical center where saliva and serum specimens were collected concurrently in dim light conditions during a 14-hr overnight study period and analyzed for melatonin levels with radioimmunoassays (RIAs). Eighty-five subjects over the age of 65 with a broad range of medical conditions participated in the study. Subjects with dementia, depression and anemia were excluded. We found that saliva volume was inadequate for analysis (<200 microL) in 23.6% of specimens, with the majority of inadequate volume specimens occurring after midnight and inadequate specimens occurring more frequently in females than in males. The correlation coefficient for saliva melatonin and serum melatonin was r = 0.659 (Spearman, P < 0.001), and r = 0.466 for saliva dim light melatonin onset (DLMO) and serum DLMO. Saliva melatonin levels were 30.9% of serum melatonin levels, with a wide range of ratios noted between subjects. Overall melatonin levels influenced both the correlation and ratio of saliva melatonin to serum melatonin; higher correlations and lower ratios were noted when melatonin levels were high. Saliva specimens provide an economical and practical method for melatonin assessment, however, in older adults, issues such as hyposalivation and low melatonin levels limit the feasibility and validity, respectively, of saliva melatonin. PMID- 12562500 TI - Melatonin attenuates kainic acid-induced hippocampal neurodegeneration and oxidative stress through microglial inhibition. AB - The antioxidant and anti-inflammatory effects of melatonin on kainic acid (KA) induced neurodegeneration in the hippocampus were evaluated in vivo. It has been suggested that the pineal secretory product, melatonin, protects neurons in vitro from excitotoxicity mediated by kainate-sensitive glutamate receptors, and from oxidative stress-induced DNA damage and apoptosis. In this study, we injected 10 mg/kg kainate intraperitoneally (i.p.) into adult male Sprague-Dawley rats. This results in selective neuronal degeneration accompanied by intense microglial activation and triggers DNA damage in the hippocampus. We tested the in vivo efficacy of melatonin in preventing KA-induced neurodegeneration, oxidative stress and neuroinflammation in the hippocampus. Melatonin (2.5 mg/kg, i.p.) was given 20 min before, immediately after, and 1 and 2 hr after KA administration. Rats were killed 72 hr later and their hippocampi were examined for evidence of DNA damage (in situ dUTP end-labeling, i.e. TUNEL staining), cell viability (hematoxylin and eosin staining), and microglial (isolectin-B4 histochemistry) and astroglial responses (glial fibrillary acidic protein immunohistochemistry), as well as lipid peroxidation (4-hydroxynonenal immunohistochemistry). A cumulative dose of 10 mg/kg melatonin attenuates KA-induced neuronal death, lipid peroxidation, and microglial activation, and reduces the number of DNA breaks. A possible mechanism for melatonin-mediated neuroprotection involves its antioxidant and anti-inflammatory actions. The present data suggest that melatonin is potentially useful in the treatment of acute brain pathologies associated with oxidative stress-induced neuronal damage such as epilepsy, stroke, and traumatic brain injury. PMID- 12562501 TI - Norepinephrine-dependent phosphorylation of the transcription factor cyclic adenosine monophosphate responsive element-binding protein in bovine pinealocytes. AB - Norepinephrine (NE)-dependent activation of transcription factors is of central importance for the rhythmic production of melatonin in the rodent pineal gland. At variance with rodents, NE regulates melatonin biosynthesis through post translational mechanisms in ungulates, and it is not yet known whether transcription factors play any role in ungulate pineal functions. Here, we investigated in isolated bovine pinealocytes the NE-dependent phosphorylation of the transcription factor cyclic adenosine monophosphate (cAMP) responsive element binding protein (CREB) and compared the effects of NE with those of vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP). Treatment with 10(-7) m NE for 30 min induced a strong nuclear phosphorylated CREB (pCREB) immunoreaction in cells that were identified as pinealocytes by immunocytochemical demonstration of serotonin, a pinealocyte specific marker. Immunoblots showed that the NE-induced immunoreaction was due to phosphorylation of the transcription factor CREB and another protein, presumably the activating transcription factor 1 (ATF-1). 10(-7) m isoproterenol (ISO) or 10(-5) m forskolin mimicked the response to NE indicating that NE acts through the beta-adrenergic/cAMP pathway. Also 10(-7) m PACAP, but not 10(-7) m VIP enhanced CREB phosphorylation; however, only a subpopulation of cells was responsive to PACAP. Our results suggest that, irrespective of whether or not melatonin production is controlled via transcriptional mechanisms, NE-induced CREB phosphorylation represents a very conserved element in pineal physiology of mammals because NE increases pCREB levels in all mammalian species investigated so far. However, the genes targeted by pCREB may vary from one mammalian species to the other. Our results also suggest that transcription factors other than pCREB, like ATF-1, may play a role in pineal functions of mammals. PMID- 12562502 TI - Melatonin reduces nitric oxide level during ischemia but not blood-brain barrier breakdown during reperfusion in a rat middle cerebral artery occlusion stroke model. AB - Melatonin is a potent antioxidant and free radical scavenger. Previously, we showed that a single injection of melatonin before ischemia significantly reduced the infarct volume in both permanent and 3-hr middle cerebral artery occlusion (MCAO) rat stroke models. Nitric oxide (NO) and other free radicals play an important role in the pathogenesis of cerebral ischemia, and they have been postulated to mediate the breakdown of the blood-brain barrier (BBB) during ischemia. In this study, we evaluated the influence of melatonin, given at 30 min before MCAO, on brain NO concentration and BBB breakdown. Brain NO concentration was measured at 15 min of MCAO using electron paramagnetic resonance spectroscopy. BBB breakdown at 3 hr of reperfusion following 3 hr of MCAO was assessed using Evans blue extravasation. The relative brain NO concentration was increased to 141.69 +/- 9.71% (mean +/- S.E.M.; n = 9) at 15 min of MCAO. Treatment with melatonin at 1.5, 5, or 50 mg/kg significantly reduced the brain NO concentration to 104.20 +/- 11.20% (n = 8), 55.67 +/- 5.58% (n = 11), and 104.86 +/- 12.56% (n = 9), respectively. Melatonin at 5 mg/kg did not affect Evans blue extravasation. Our results suggest that a single injection of melatonin protects against focal cerebral ischemia partly via inhibition of ischemia-induced NO production and that this regimen does not prevent BBB breakdown following ischemia-reperfusion. PMID- 12562503 TI - Melatonin exerts a therapeutic effect on cholestatic liver injury in rats with bile duct ligation. AB - We examined whether melatonin exerts a therapeutic effect on cholestatic liver injury in rats treated with bile duct ligation (BDL). Cholestatic liver injury was induced in male Wistar rats aged 4 wk by ligating the bile duct. Cholestatic liver injury developed 5 days after BDL and continued to 13 days, judging from the levels of serum hepatobiliary injury markers. The serum concentration of thiobarbituric acid reactive substances (TBARS), an index of lipid peroxidation, and the hepatic level of TBARS and the activity of hepatic myeloperoxidase, an index of tissue neutrophil infiltration, increased 5 days after BDL, and these increases were enhanced at 13 days. A similar increase in the serum total cholesterol concentration occurred 5 and 13 days after BDL, while the hepatic cholesterol concentration tended to increase at 13 days. When melatonin [10 or 100 mg/kg body weight (BW)] was orally administered to BDL-treated rats everyday for 8 days, starting 5 days after BDL, the indoleamine attenuated cholestatic liver injury observed at 13 days after BDL was more effective at the higher dose than at the lower dose. The administered melatonin (10 or 100 mg/kg BW) reduced the increases in serum and hepatic TBARS concentrations and hepatic myeloperoxidase activity observed at 13 days after BDL and the higher dose of indoleamine was more effective than the lower dose. Neither dose of melatonin affected the increased serum total cholesterol concentration or the hepatic cholesterol concentration observed at 13 days after BDL. These results indicate that orally administered melatonin at pharmacological doses exerts a therapeutic effect on cholestatic liver injury in rats with BDL possibly through its antioxidant and anti-inflammatory actions. PMID- 12562504 TI - Suppression of hypothalamic pro-opiomelanocortin (POMC) gene expression by daily melatonin supplementation in aging rats. AB - Both plasma melatonin levels and hypothalamic arcuate nucleus pro opiomelanocortin (POMC) (biosynthetic precursor to the endogenous opioid ss endorphin and other opiomelanocortins) mRNA content decrease with aging. To test whether the decline in melatonin is responsible for the decline in POMC mRNA, we investigated the effects of daily melatonin treatment on hypothalamic POMC mRNA content in middle-aged and older Sprague-Dawley rats. Daily nocturnal melatonin treatment (50 microg kg bw(-1) night(-1), in the night-time drinking water) for 7 months, starting at 13 months of age, did not significantly alter female arcuate nucleus POMC mRNA content determined at the end of the light period (i.e., before nightly melatonin administration), but suppressed (24%, P < 0.05) POMC mRNA content at the end of the dark period (i.e., following melatonin administration). Likewise, nocturnal administration of 50 or 500 microg melatonin kg bw(-1) night( 1) to male rats for 7 months suppressed (31 or 28%, respectively; P < 0.05) POMC mRNA content at the middle of the dark period at 20 months of age. Finally, 10 wk administration of 30 microg melatonin kg bw(-1) day(-1) suppressed (31%, P < 0.01) POMC mRNA content in middle-aged male rats killed at the end of the dark period. Melatonin treatments did not significantly alter estradiol or testosterone levels. Thus, moderate-dosage nocturnal melatonin supplementation suppressed nocturnal hypothalamic POMC gene expression in both middle-aged males and females, suggesting that melatonin supplementation during aging decreases, rather than increases, forebrain opiomelanocortinergic activity. These POMC responses were apparently not dependent on gonadal steroid responses and did not become refractory to melatonin treatment maintained until old age. PMID- 12562505 TI - Some biochemical properties of melatonin and the characterization of a relevant metabolite arising from its interaction with H2O2. AB - Melatonin is an efficient protector against hydrogen peroxide(H2O2)-induced lipid peroxidation and acts in a concentration-dependent manner. Hydrogen peroxide is rather a water stable molecule which is able to cross the cell membrane much better than some important free radicals such as superoxide anion, and consequently its local production can lead to significant spread by diffusion. In this paper we report data regarding some biochemical properties of melatonin as well as the chemical characterization of the major product formed from the interaction between melatonin and H2O2 (N1-acetyl-N2-formyl-5-methoxykynuramine) that are consistent with previous data reported by other authors. The effect of melatonin on catalase, glutathione peroxidase and superoxide dismutase in in vitro and in vivo experiments is also reported. PMID- 12562506 TI - Melatonin increases oestradiol-induced bone formation in ovariectomized rats. AB - To assess the effect of melatonin on bone metabolism in ovariectomized rats, receiving oestradiol therapy or not, melatonin was administered in the drinking water (25 microg/mL water) and oestradiol (10 microg/kg body weight) or vehicle was given subcutaneously 5 days/week for up to 60 days after surgery. Urinary deoxypyridinoline (a marker of bone resorption) and circulating levels of bone alkaline phosphatase activity (a marker of bone formation), as well as serum calcium and phosphorus levels, were measured every 15 days. Bone area (BA), bone mineral content (BMC), bone mineral density (BMD) and total body fat (expressed as 100 g body weight) were measured by dual-energy X-ray absorptiometry at the end of the experiment. Body weight and total body fat were augmented after ovariectomy, and decreased after melatonin or oestradiol treatment. The effect of melatonin on body weight was seen in sham-operated rats only. Ovariectomy augmented, and melatonin or oestradiol lowered, urinary deoxypyridinoline excretion. This effect of melatonin and oestradiol was seen mainly in ovariectomized rats. The efficacy of oestradiol to counteract ovariectomy-induced bone resorption was increased by melatonin. Melatonin or oestradiol lowered serum bone alkaline phosphatase activity. Melatonin inhibition was seen mainly on the increase of bone alkaline phosphatase activity that followed ovariectomy. Serum phosphorus levels decreased after melatonin administration and were augmented after oestradiol injection; overall, melatonin impaired the increase of serum phosphorus caused by oestradiol. Ovariectomy decreased, and oestradiol increased, serum calcium levels while melatonin augmented serum calcium in sham-operated rats only. On day 60 after surgery, BMD and content decreased after ovariectomy and were increased after oestradiol injection. Melatonin augmented BA of spine and BMC of whole of the skeleton and tibia. The highest values observed were those of rats treated concurrently with oestradiol and melatonin. The present results indicate that: (i) melatonin treatment restrained bone remodelling after ovariectomy; (ii) the effect of melatonin required adequate concentrations of oestradiol; (iii) melatonin augmented oestradiol effects on bone in ovariectomized rats; (iv) a counter-regulation by melatonin of the increase in body fat caused by ovariectomy was uncovered. The melatonin doses employed were pharmacological in terms of circulating melatonin levels but not necessarily for some other fluids or tissues. PMID- 12562507 TI - Somatostatin receptors (sst2) are coupled to Go and modulate GTPase activity in the rabbit retina. AB - The role of somatostatin and its mechanism of action in the retina remains an important target for investigation. Biochemical and pharmacological studies were engaged to characterize the somatostatin receptors in the rabbit retina, and their coupling to G-proteins. The ability of selective ligands to inhibit [125I]Tyr11-somatostatin-14 binding to rabbit retinal membranes was examined. The sst2 analogues SMS201-995, MK678, and BIM23014, displayed IC50 values of 0.28 +/- 0.12, 0.04 +/- 0.01 and 1.57 +/- 0.39 nm, respectively. The sst1 analogue CH275 moderately displaced the [125I]Tyr11-somatostatin-14 binding, while selective analogues for sst3, sst4 and sst5 had minimal effect. Immunoblotting and/or immunohistochemistry studies revealed the presence of the pertussis toxin sensitive Gi1/2, and Go proteins, as well as Gs. Somatostatin-14 and MK678 stimulated GTPase activity in a concentration-dependent manner with EC50 values of 42.8 +/- 16.8 and 70.0 +/- 16.5 nm, respectively, thus supporting the functional coupling between the receptor and the G-proteins. CH275 stimulated the GTPase activity moderately, in agreement with its binding profile. The antisera raised against Goalpha and Gi1/2alpha inhibited the somatostatin-induced high affinity GTPase activity, but only anti-Goalpha inhibited the MK678 stimulation of the enzyme. These results suggest that somatostatin mediates its actions in the rabbit retina by interacting mainly with sst2 receptors that couple to Goalpha. PMID- 12562508 TI - Inhibition of NADH oxidation by chloramphenicol in the freely moving rat measured by picosecond time-resolved emission spectroscopy. AB - Owing to the lack of methods capable to monitor the energetic processes taking place within small brain regions (i.e. nucleus raphe dorsalis, nRD), the neurotoxicity of various categories of substances, including antibiotics and psycho-active drugs, still remains difficult to evaluate. Using an in vivo picosecond optical spectroscopy imaging method, we report that chloramphenicol (CAP), besides its well-known ability to inhibit the mitochondria protein synthesis, also influences the NADH/NAD+ redox processes of the respiratory chain. At a 200-mg/kg dose, CAP indeed produces a marked increase in the fluorescent signal of the nRD which, according to clear evidence, is likely to be related to the NADH concentration. This effect also implies an efficient inhibition of complex I of the respiratory chain by CAP. It refers to the mechanism through which the adverse effects of the antibiotic may take place. It could explain why paradoxical sleep, a state needing aerobic energy to occur, is suppressed after CAP administration. The present approach constitutes the first attempt to determine by fluorescence methods the effects of substances on deep brain structures of the freely moving animal. It points out that in vivo ultrafast optical methods are innovative and adequate tools for combined neurochemical and behavioural approaches. PMID- 12562509 TI - Ceramide increases mitochondrial free calcium levels via caspase 8 and Bid: role in initiation of cell death. AB - We investigated how the mitochondrial phase of ceramide-mediated cell death is initiated in nerve growth factor (NGF)-differentiated PC12 cells. We distinguished three independent effects of ceramide: free radical production; a transient increase in cytosolic free calcium; and a long-lasting increase in mitochondrial free calcium. Only the latter led to cell death, which could be prevented by buffering of mitochondrial calcium with the calcium binding protein calbindin D-28K ectopically expressed in mitochondria. We showed that mitochondrial calcium did not increase as a result of the increase in cytosolic free calcium levels. Rather, it appears to derive from the endoplasmic reticulum (ER) since dantrolene, which inhibits release of calcium from ER into cytosol through ryanodine receptors, prevented the increase in cytosolic free calcium but potentiated the increase in mitochondrial free calcium. This suggests that a transfer of calcium occurs directly, or very locally, between the two organelles. This transfer implicated activation of caspase 8 and cleavage of its substrate Bid, a previously unknown function of these cell death intermediaries. The increase in mitochondrial free calcium was also responsible for the release of cytochrome c into the cytosol, underlining the critical role it plays in ceramide mediated cell death. PMID- 12562511 TI - Transmitter uptake and release in PC12 cells overexpressing plasma membrane monoamine transporters. AB - Transmitter uptake and exocytosis of secretory vesicles are two essential aspects of neurotransmission. Here we show that transient overexpression of plasma membrane monoamine transporters in rat pheochromocytoma PC12 cells induced an approximate 20-fold enhancement of cellular uptake of monoamines. Intravesicular amine concentration was greatly increased, as demonstrated directly by carbon fibre amperometry. However, the amount of stored monoamines diminished over a 5-h period, unless monoamine oxidase was inhibited, indicating that monoamines leak out from secretory vesicles. This efflux of monoamines accounts for the reported dependence of vesicular monoamine content (the quantal size) on the kinetics of vesicular monoamine uptake. Measuring radiolabelled monoamines release from the cell population provided accurate determination of the secretory activity of the subpopulation (10-20%) of cells transfected with monoamine transporters, since they contained about 95% of the radiolabel. Accordingly, significant modification of the secretory responses was observed, at the cell population level, upon transient expression of the serotonin transporter and of proteins known to interfere with exocytosis, such as botulinum neurotoxin C1, GTPase-deficient Rab3 proteins, truncated Rabphilin constructs or Rim. The co-transfection assay described here, based on transient expression of monoamine transporters, should prove useful in functional studies of the secretory machinery. PMID- 12562510 TI - Palmitic and stearic fatty acids induce caspase-dependent and -independent cell death in nerve growth factor differentiated PC12 cells. AB - Apoptotic cell death has been proposed to play a role in the neuronal loss observed following traumatic injury in the CNS and PNS. The present study uses an in vitro tissue culture model to investigate whether free fatty acids (FFAs), at concentrations comparable to those found following traumatic brain injury, trigger cell death. Nerve growth factor (NGF)-differentiated PC12 cells exposed to oleic and arachidonic acids (2 : 1 ratio FFA/BSA) showed normal cell survival. However, when cells were exposed to stearic and palmitic acids, there was a dramatic loss of cell viability after 24 h of treatment. The cell death induced by stearic acid and palmitic acid was apoptotic as assessed by morphological analysis, and activation of caspase-8 and caspase-3-like activities. Western blotting showed that differentiated PC12 cells exposed to stearic and palmitic acids exhibited the signature apoptotic cleavage fragment of poly (ADP-ribose) polymerase (PARP). Interestingly, blockade of caspase activities with the pan caspase inhibitor z-VAD-fmk failed to prevent the cell death observed induced by palmitic or stearic acid. RT-PCR and RNA blot experiments showed an up-regulation of the Fas receptor and ligand mRNA. These findings are consistent with our hypothesis that FFAs may play a role in the cell death associated with trauma in the CNS and PNS. PMID- 12562512 TI - The basic helix-loop-helix differentiation factor Nex1/MATH-2 functions as a key activator of the GAP-43 gene. AB - Nex1/MATH-2 is a neurogenic basic Helix-Loop-Helix (bHLH) transcription factor that belongs to the NeuroD subfamily. Its expression parallels that of the GAP-43 gene and peaks during brain development, when neurite outgrowth and synaptogenesis are highly active. We previously observed a direct correlation between the levels of expression of Nex1 and GAP-43 proteins, which resulted in extensive neurite outgrowth and neuronal differentiation of PC12 cells in the absence of nerve growth factor. Since the GAP-43 gene is a target for bHLH regulation, we investigated whether Nex1 could regulate the activity of the GAP 43 promoter. We found that among the members of the NeuroD subfamily, Nex1 promoted maximal activity of the GAP-43 promoter. The Nex1-mediated activity is restricted to the conserved E1-E2 cluster located near the major transcription start sites. By electrophoretic mobility shift assay and site-directed mutagenesis, we showed that Nex1 binds as homodimers and that the E1 E-box is a high affinity binding site. We further found that Nex1 released the ME1 E-protein mediated repression in a concentration dependent manner. Thus, the E1-E2 cluster has a dual function: it can mediate activation or repression depending on the interacting bHLH proteins. Finally, a series of N-terminal and C-terminal deletions revealed that Nex1 transcriptional activity is linked to two distinct transactivation domains, TAD1 and TAD2, with TAD1 being unique to Nex1. Together, our results suggest that Nex1 may engage in selective interactions with components of the core transcriptional machinery whose assembly is dictated by the architecture of the GAP-43 promoter and cellular environment. PMID- 12562513 TI - Far-upstream elements are dispensable for tissue-specific proenkephalin expression using a Cre-mediated knock-in strategy. AB - Several cis-regulatory DNA elements are present in the 5' upstream regulatory region of the enkephalin gene (ENK) promoter. To determine their role in conferring organ-specificity of ENK expression in mice and to circumvent the position effects from random gene insertion that are known to often frustrate such analysis in transgenic mice, we used a Cre-mediated gene knock-in strategy to target reporter constructs to a "safe haven" loxP-tagged locus in the hypoxanthine phosphoribosyltransferase (HPRT) gene. Here we report reliable and reproducible reporter gene expression under the control of the 5' upstream regulatory region of the mouse ENK gene in gene-modified mice using this Cre mediated knock-in strategy. Comparison of two 5'ENK regulatory regions (one with and the other without known cis-regulatory DNA elements) in the resulting adult mice showed that conserved far-upstream cis-regulatory DNA elements are dispensable for correct organ-specific gene expression. Thus the proximal 1.4 kb of the murine ENK promoter region is sufficient for organ-specificity of ENK gene expression when targeted to a safe-haven genomic locus. These results suggest that conservation of the far-upstream DNA elements serves more subtle roles, such as the developmental or cell-specific expression of the ENK gene. PMID- 12562514 TI - Cannabinoid CB1 receptor knockout mice exhibit markedly reduced voluntary alcohol consumption and lack alcohol-induced dopamine release in the nucleus accumbens. AB - The mechanisms underlying predisposition to alcohol abuse and alcoholism are poorly understood. In this study, we evaluated the role of cannabinoid (CB1) receptors in (i) voluntary alcohol consumption, and (ii) acute alcohol-induced dopamine (DA) release in the nucleus accumbens, using mice that lack the CB1 receptor gene (CB1-/-). CB1-/- mice exhibited dramatically reduced voluntary alcohol consumption, and completely lacked alcohol-induced DA release in the nucleus accumbens, as compared to wild-type mice. The gender difference, with female mice consuming significantly more alcohol than wild-type male mice, was observed in wild-type mice, whereas this gender difference was nonexistent in CB1 mutant male and female mice. There was also a significant gender difference, with the wild-type, heterozygous, and mutant females consuming significantly more liquid and food than wild-type, heterozygous and mutant males. However, the total volume of fluid consumption and food intake did not differ between wild-type, heterozygous, and mutant mice. These results strongly suggest that the CB1 receptor system plays an important role in regulating the positive reinforcing properties of alcohol. PMID- 12562515 TI - Novel glucocorticoid effects on acute inflammation in the CNS. AB - The CNS can mount an inflammatory reaction to excitotoxic insults that contributes to the emerging brain damage. Therefore, anti-inflammatory drugs should be beneficial in neurological insults. In contrast, glucocorticoids (GCs), while known for their anti-inflammatory effects, can exacerbate neurotoxicity in the hippocampus after excitotoxic insults. We investigated the effect of GCs on the inflammatory response after a neurological insult. Intact control (INT; intact stress response GC profile), adrenalectomized/GC-supplemented (ADX; low basal GC profile) and GC-treated (COR; chronically high GC profile) rats were injected with kainic acid into the hippocampal CA3 region. Lesion size was determined 8-72 h later. The inflammatory response was characterized using immunohistochemistry, RNAse protection assay and ELISA. The INT and COR rats developed larger CA3 lesions than ADX rats. We found that GCs surprisingly caused an increase in relative numbers of inflammatory cells (granulocytes, monocytes/macrophages and microglia). Additionally, mRNA and protein (IL-1beta and TNF-alpha) levels of the pro-inflammatory cytokines IL-1alpha, IL-1beta and TNF-alpha were elevated in COR rats compared with INT and ADX rats. These data strongly question the traditional view of GCs being uniformly anti-inflammatory and could further explain how GCs worsen the outcome of neurological insults. PMID- 12562516 TI - Rhythmic changes in metabolism of dopamine in the chick retina: the importance of light versus biological clock. AB - Rhythmic changes in dopamine (DA) content and metabolism were studied in retinas of chicks that were adapted to three different lighting conditions: 12-h light : 12-h dark (LD), constant darkness (DD) and continuous light (LL). Retinas of chicks kept under LD conditions exhibited light-dark-dependent variations in the steady-state level of DA and the two metabolites of DA, i.e. 3,4 dihydroxyphenylacetic acid (DOPAC) and homovanilic acid (HVA). Concentrations of DA, DOPAC and HVA were high in light hours and low in dark hours of the LD illumination cycle. In retinas of chicks kept under DD, the content of DA, DOPAC and HVA oscillated in a rhythmic manner for 2 days, with higher values during the subjective light phase than during the subjective dark phase. The amplitudes of the observed oscillations markedly and progressively declined compared with the amplitudes recorded under the LD cycle. In retinas of chicks kept under LL conditions, levels of DA, DOPAC and HVA were similar to those found during the light phase of the LD cycle. Changes in the retinal contents of DA and HVA did not exhibit pronounced daily oscillations, while on the first day of LL the retinal concentrations of DOPAC were significantly higher during the subjective light phase than during the subjective dark phase. Acute exposure of chicks to light during the dark phase of the LD cycle markedly increased DA and DOPAC content in the retina. In contrast, light deprivation during the day decreased the retinal concentrations of DA and DOPAC. It is suggested that of the two regulatory factors controlling the level and metabolism of DA in the retina of chick, i.e. light and biological clock, environmental lighting conditions seem to be of major importance, with light conveying a stimulatory signal for the retinal dopaminergic cells. PMID- 12562517 TI - Mechanisms by which 2',3'-dideoxyinosine (ddI) crosses the guinea-pig CNS barriers; relevance to HIV therapy. AB - The influence of transport mechanisms at the blood-brain barrier (BBB) and blood CSF barrier (choroid plexus) on the CNS distribution of anti-human immunodeficiency virus (HIV) drugs was examined using guinea-pig brain perfusion and incubated choroid plexus models. 2',3'-dideoxyinosine (ddI) passage across the BBB was demonstrated to be via non-saturable (Kd = 0.22 +/- 0.3 microL/min/g) and saturable (Km = 20.1 +/- 15.0 microm, Vmax = 6.5 +/- 2.1 pmol/min/g) processes. Cross competition studies implicated an equilibrative nucleoside transporter in this influx. The brain distribution of ddI was unchanged in the presence of additional nucleoside reverse transcriptase inhibitors (NRTIs). ddI transport from blood into choroid plexus was demonstrated to involve an organic anion transporting polypeptide 2-like transporter. The NRTIs, abacavir, 3'-azido 3'-deoxythymidine and (-)-beta-L-2',3'-dideoxy-3'-thiacytidine, competed with ddI for transporter binding sites at the choroid plexus, altering the tissue concentration of ddI. This has clinical implications as the choroid plexus is a site of HIV replication, and suboptimal CNS concentrations of anti-HIV drugs could result in neurological complications. Furthermore, this may promote the selection of drug resistant variants of HIV within the CNS, which could re-infect the periphery and lead to HIV therapy failure. This study indicates that understanding drug interactions at the transporter level could prove valuable when selecting drug combinations to treat HIV within the CNS. PMID- 12562518 TI - Functional analysis of connexin-26 mutants associated with hereditary recessive deafness. AB - The physiological importance of connexin-26 (Cx26) gap junctions in regulating auditory function is indicated by the finding that autosomal recessive DFNB1 deafness is associated with mutations of the Cx26 gene. To investigate the pathogenic role of Cx26 mutation in recessive hearing loss, four putative DFNB1 Cx26 mutants (V84L, V95M, R127H, and R143W) were stably expressed in N2A cells, a communication-deficient cell line. In N2A cells expressing (R127H) Cx26 gap junctions, macroscopic junctional conductance and ability of transferring neurobiotin between transfected cells were greatly reduced. Despite the formation of defective junctional channels, immunoreactivity of (R127H) Cx26 was mainly localized in the cell membrane and prominent in the region of cell-cell contact. Mutant (V84L), (V95M), or (R143W) Cx26 protein formed gap junctions with a junctional conductance similar to that of wild-type Cx26 junctional channels. (V84L), (V95M), or (R143W) Cx26 gap junctions also permitted neurobiotin transfer between pairs of transfected N2A cells. The present study suggests that (R127H) mutation associated with hereditary sensorineural deafness results in the formation of defective Cx26 gap junctions, which may lead to the malfunction of cochlear gap junctions and hearing loss. Further studies are required to determine the exact mechanism by which mutant (V84L), (V95M), and (R143W) Cx26 proteins, which are capable of forming functional homotypic junctional channels in N2A cells, cause the cochlear dysfunction and sensorineural deafness. PMID- 12562519 TI - GABAB receptor mRNA in the raphe nuclei: co-expression with serotonin transporter and glutamic acid decarboxylase. AB - We have used double-label in situ hybridization techniques to examine the cellular localization of GABAB receptor mRNA in relation to serotonin transporter mRNA and glutamic acid decarboxylase mRNA in the rat dorsal raphe, median raphe and raphe magnus nuclei. The degree of cellular co-localization of these markers notably varied among the different nuclei. In the dorsal raphe, cell bodies showing GABAB receptor mRNA were very abundant, the 85% being also labelled for serotonin transporter mRNA, and a low proportion (5%) showing glutamic acid decarboxylase mRNA. In the median raphe, the level of co-expression of GABAB receptor mRNA with serotonin transporter mRNA was significantly lower. Some cells were also identified that contained GABAB receptor mRNA in the absence of either one of the other mRNA species studied. Our results support the presence of GABAB receptors in serotonergic as well as GABAergic neurones in the dorsal and median raphe, providing the anatomical basis for the reported dual inhibitory/disinhibitory effect of the GABAB agonist baclofen on serotonergic function. PMID- 12562520 TI - Nicotine trapping causes the persistent desensitization of alpha4beta2 nicotinic receptors expressed in oocytes. AB - To determine whether prolonged nicotine exposure persistently inactivates rat alpha4beta2 nicotinic receptors expressed in Xenopus oocytes, we measured the voltage-clamped alpha4beta2 response to acetylcholine (ACh) before and 24 h after, 1-h or 12-h incubations in 10 microm nicotine. A 12-h incubation in 10 microm nicotine depressed the alpha4beta2 ACh response for 24 h without affecting total or surface alpha4beta2 expression. To determine whether oocyte-mediated nicotine release caused this depression, we co-incubated an alpha4beta2 expressing oocyte with an un-injected one (pre-incubated in 10 microm nicotine for 12 h) for 24 h and measured the change in the alpha4beta2 ACh response. The response decreased by the same factor after the co-incubation as it did after a 12-h incubation in 10 microm nicotine and a 24-h incubation in nicotine-free media. Thus, oocyte-mediated nicotine release caused the persistent desensitization we observed after a 12-h incubation in 10 microm nicotine. Consistent with this result, measurements of [3H]nicotine release show that oocytes release enough nicotine into the wash media to desensitize alpha4beta2 receptors and that prolonged incubation in 300 microm ACh (which cannot readily cross the membrane or accumulate in acidic vesicles) did not persistently depress the alpha4beta2 response. PMID- 12562521 TI - Possible regulation by N-methyl-d-aspartate receptors of proliferative progenitor cells expressed in adult mouse hippocampal dentate gyrus. AB - An immunohistochemical technique was employed to analyze mechanisms underlying modulation by N-methyl-d-aspartate (NMDA) receptors of proliferation of neural progenitor cells in adult mouse brain. The systemic administration of NMDA at 100 mg/kg resulted in marked expression of c-Fos, Fra-2 and c-Jun proteins in the granule cell layers of the dentate gyrus in murine hippocampus 2 h later, followed by a significant reduction of the incorporation of 5-bromo-2' deoxyuridine (BrdU) in a manner sensitive to the antagonist dizocilpine 2 days after administration. The administration of NMDA also suppressed constitutive expression of both nestin and proliferating cell nuclear antigen (PCNA) in the dentate granule cells 2 days later, without markedly affecting cell viability for up to 8 weeks after administration. In the subventricular zone and olfactory bulb, however, NMDA failed to affect either the incorporation of BrdU or the expression of nestin and PCNA. The NR1 subunit was highly expressed in the dentate gyrus in addition to the stratum oriens in the hippocampus, but not in the subventricular zone and olfactory bulb. These results suggest that NMDA receptors may play a role crucial for maintenance of the integrity and function of proliferative neural progenitor cells through expression of the nuclear transcription factor activator protein-1 in granule cells of the dentate gyrus in adult mouse brain. PMID- 12562522 TI - Altered cAMP-dependent protein kinase subunit immunolabeling in post-mortem brain from patients with bipolar affective disorder. AB - Previous findings of reduced [3H]cAMP binding and increased activities of cAMP dependent protein kinase (PKA) in discrete post-mortem brain regions from patients with bipolar affective disorder (BD) suggest that PKA, the major downstream target of cAMP, is also affected in this illness. As prolonged elevation of intracellular cAMP levels can modify PKA regulatory (R) and catalytic (C) subunit levels, we sought to determine whether these PKA abnormalities are related to changes in the abundance of PKA subunits in BD brain. Using immunoblotting techniques along with PKA subunit isoform-specific polyclonal antisera, levels of PKA RIalpha, RIbeta, RIIalpha, RIIbeta and Calpha subunits were measured in cytosolic and particulate fractions of temporal, frontal and parietal cortices of post-mortem brain from BD patients and matched, non-neurological, non-psychiatric controls. Immunoreactive levels of cytosolic Calpha in temporal and frontal cortices, as well as that of cytosolic RIIbeta in temporal cortex, were significantly higher in the BD compared with the matched control brains. These changes were independent of age, post-mortem interval or pH and unrelated to ante-mortem lithium treatment or suicide. These findings strengthen further the notion that the cAMP/PKA signaling system is up-regulated in discrete cerebral cortical regions in BD. PMID- 12562523 TI - Plasticity of glutamatergic control of striatal acetylcholine release in experimental parkinsonism: opposite changes at group-II metabotropic and NMDA receptors. AB - To investigate whether adaptive changes of glutamatergic transmission underlie dysfunction of the cholinergic system in experimental parkinsonism, the effects of group-II metabotropic glutamate and NMDA receptor ligands on acetylcholine release was studied in striatal slices and synaptosomes obtained from naive rats, 6-hydroxydopamine hemi-lesioned rats and 6-hydroxydopamine hemi-lesioned rats chronically treated with levodopa (L-DOPA) plus benserazide (non-dyskinetic). Group-II metabotropic glutamate receptor agonists LY354740, DCG-IV and L-CCG-I inhibited the electrically-evoked endogenous acetylcholine release from slices, while NMDA facilitated it. LY354740 also inhibited K+-evoked acetylcholine release from synaptosomes. LY354740-induced inhibition was prevented by the group II metabotropic glutamate receptor antagonist LY341495. In hemi-parkinsonian rats, sensitivity towards LY354740 was reduced while that to NMDA was enhanced in the lesioned (denervated) compared with unlesioned striatum. Moreover, dizocilpine inhibited acetylcholine release in the lesioned compared with unlesioned striatum. Chronic treatment with L-DOPA normalized sensitivity towards glutamatergic agonists. We conclude that striatal dopamine denervation results in plastic changes at group-II metabotropic glutamate and NMDA receptors that may shift glutamatergic control of acetylcholine release towards facilitation. From a clinical perspective, L-DOPA and NMDA antagonists appear effective in counteracting overactivity of striatal cholinergic interneurones associated with Parkinson's disease. PMID- 12562524 TI - Regulation of alpha-synuclein by bFGF in cultured ventral midbrain dopaminergic neurons. AB - Alpha-synuclein is a neuronal protein that is implicated in the control of synaptic vesicle function and in Parkinson's disease (PD). Consequently, alterations of alpha-synuclein levels may play a role in neurotransmission and in PD pathogenesis. However, the factors that regulate alpha-synuclein levels are unknown. Growth factors mediate neurotrophic and plasticity effects in CNS neurons, and may play a role in disease states. Here we examine the regulation of alpha-synuclein levels in primary CNS neurons, with particular emphasis on dopaminergic neurons. E18 rat cortical neurons and dopaminergic neurons of E14 rat ventral midbrain showed an induction of alpha-synuclein protein levels with maturation in culture. Application of basic Fibroblast growth factor (bFGF) promoted alpha-synuclein expression selectively within dopaminergic, and not GABAergic or cortical neurons. This induction was blocked by actinomycin D, but not by inhibition of bFGF-induced glial proliferation. alpha-Synuclein levels were not altered by glial-derived neurotrophic factor (GDNF), or by apoptotic stimuli. We conclude that bFGF promotes alpha-synuclein expression in cultured ventral midbrain dopaminergic neurons through a direct transcriptional effect. These results suggest that distinct growth factors may thus mediate plasticity responses or influence disease states in ventral midbrain dopaminergic neurons. PMID- 12562525 TI - Selective E2F-dependent gene transcription is controlled by histone deacetylase activity during neuronal apoptosis. AB - The alteration of chromatin through histone acetylation and deacetylation participates in the regulation of gene expression. We have investigated the effects of histone deacetylase inhibition on neuronal fate. We show that treatment of primary neurones with trichostatin A (TSA) or sodium butyrate (NaBu) induces typical features of apoptosis, a cell death that relies on specific genetic programmes. We have further explored the molecular mechanisms implicated in the TSA response and demonstrated that TSA-induced apoptosis is partly dependent on the activation of the transcription factor E2F-1, which has pro apoptotic functions in these neurones. Furthermore, the increased e2f-1 transcriptional response is probably the result of mechanisms occurring through E2F-responsive elements. Histone acetylation also takes place at the e2f-1 promoter, but this modification is neither required nor by itself sufficient to induce increased transcription at the e2f-1 promoter. Activation might thus occur through acetylation of non-histone proteins binding this regulatory element. Finally, we show that TSA induces the transcription of E2F-dependent genes, such as its cell cycle target cyclin E, but also pro-apoptotic genes, such as Apaf1. Taken together, our results suggest that, in neuroprotective conditions, histone deacetylase activity allows a constitutive repression of the e2f-1 gene in mature neurones in order to ensure survival. Deregulation of this repression will ultimately lead to an E2F-dependent cell death. PMID- 12562526 TI - Cloning and characterization of a novel variant (mM-rdgBbeta1) of mouse M-rdgBs, mammalian homologs of Drosophila retinal degeneration B gene proteins, and its mRNA localization in mouse brain in comparison with other M-rdgBs. AB - We report the cloning, characterization and localization in the brain of a novel isoform termed mM-rdgBbeta1 (mouse type of mammalian retinal degeneration Bbeta1 protein) in comparison with the localization of three known mammalian homologs (M rdgBbeta, M-rdgB1, M-rdgB2). mM-rdgBbeta1 cDNA contains a sequence of 119 bp as a form of insertion in the open reading frame of the known mM-rdgBbeta, and encodes a protein of 269 amino acids with a calculated molecular mass of 31.7 kDa, different from the molecular mass of 38.3 kDa of mM-rdgBbeta. It also contains a phosphatidylinositol transfer protein (PITP)-like domain similar to the known three homologs, as well as D-rdgB. The recombinant mM-rdgBbeta1 protein shows the specific binding activity to phosphatidylinositol but not to other phospholipids. This novel molecule is localized not only in the cytoplasm but also in the nucleus, different from the cytoplasmic localization of mM-rdgBbeta. In in situ hybridization analysis, the gene expression for mM-rdgBbeta1 in the brain, though weak, is rather confined to the embryonic stage, different from wider expression of mM-rdgBbeta in the gray matters of pre- and post-natal brains. Taken together, mM-rdgBbeta1 is suggested to play a role in the phosphoinositide-mediated signaling in the neural development. PMID- 12562527 TI - New Otx2 mRNA isoforms expressed in the mouse brain. AB - The mouse Otx2 gene is essential throughout head and brain development, from anterior-posterior polarity determination and neuroectoderm induction to post natal sensory organ maturation. These numerous activities must rely on a very finely tuned regulation of expression. In order to understand the molecular control of the Otx2 gene, we set out to isolate its promoter. During this quest, we identified three remote transcription start sites, two defining two new upstream exons and one mapping within the previously reported first exon. The three transcripts differed in their 5' non-coding region but encoded the same protein. The transcription start nucleotides of each mRNA species have been mapped by RNase protection assays and by an RNA circularization technique. We have demonstrated that they are all used and linked to functional promoters. In addition to leader versatility, we also detected alternative splicing within the coding sequence that gives rise to a new protein endowed with an 8 amino-acid insertion upstream of the homeodomain. Combined analysis of the relative abundance of Otx2 mRNA isoforms in representative tissues and in situ hybridization studies revealed distinct spatial and temporal, although partially overlapping, expression patterns of the mRNA isoforms. These findings provide new clues to a better understanding of the relationships between Otx2 gene architecture and its complex regulatory requirements. PMID- 12562528 TI - Human substance P receptor lacking the C-terminal domain remains competent to desensitize and internalize. AB - Substance P receptor (SPR) and its naturally occurring splice-variant, lacking the C-terminal tail, are found in brain and spinal cord. Whether C-terminally truncated SPR desensitizes like full-length SPR is controversial. We used a multivaried approach to determine whether human SPR (hSPR) and a C-terminally truncated mutant, hSPRDelta325, differ in their desensitization and internalization. In HEK-293 cells expressing either hSPRDelta325 or hSPR, SP induced desensitization of the two receptors was similar when measured by inositol triphosphate accumulation or by transient translocation of coexpressed PKCbetaII-GFP to the plasma membrane. Moreover, translocation of beta-arrestin 1 or 2-GFP (betaarr1-GFP or betaarr2-GFP) to the plasma membrane, and receptor internalization were also similar. However, hSPR and hSPRDelta325 differ in their phosphorylation and in their ability to form beta-arrestin-containing endocytic vesicles. Unlike hSPR, hSPRDelta325 is not phosphorylated to a detectable level in intact HEK293 cells, and whereas hSPR forms vesicles containing either betaarr1-GFP or betaarr2-GFP, hSPRDelta325 does not form any vesicles with betaarr1-GFP, and forms fewer vesicles with betaarr2-GFP. We conclude that truncated hSPR undergoes agonist-dependent desensitization and internalization without detectable receptor phosphorylation. PMID- 12562530 TI - Essential role for integrin linked kinase in Akt-mediated integrin survival signaling in hippocampal neurons. AB - Activation of integrin receptors in neurons can promote cell survival and synaptic plasticity, but the underlying signal transduction pathway(s) is unknown. We report that integrin signaling prevents apoptosis of embryonic hippocampal neurons by a mechanism involving integrin-linked kinase (ILK) that activates Akt kinase. Activation of integrins using a peptide containing the amino acid sequence EIKLLIS derived from the alpha chain of laminin protected hippocampal neurons from apoptosis induced by glutamate or staurosporine, and increased Akt activity in a beta1 integrin-dependent manner. Transfection of neurons with a plasmid encoding dominant negative Akt blocked the protective effect of the integrin-activating peptide, as did a chemical inhibitor of Akt. Although inhibitors of phosphoinositide-3 (PI3) kinase blocked the protective effect of the peptide, we found no increase in PI3 kinase activity following integrin stimulation suggesting that PI3 kinase was necessary for Akt activity but was not sufficient for the increase in Akt activity following integrin activation. Instead, we show a requirement for ILK in integrin receptor-induced Akt activation. ILK was activated following integrin stimulation and dominant negative ILK blocked integrin-mediated Akt activation and cell survival. Activation of ILK and Akt were also required for neuroprotection by substrate associated laminin. These results establish a novel pathway that signals cell survival in neurons in response to integrin receptor activation. PMID- 12562529 TI - Involvement of c-Jun N-terminal kinase in amyloid precursor protein-mediated neuronal cell death. AB - Amyloid precursor protein (APP), the precursor of Abeta, has been shown to function as a cell surface receptor that mediates neuronal cell death by anti-APP antibody. The c-Jun N-terminal kinase (JNK) can mediate various neurotoxic signals, including Abeta neurotoxicity. However, the relationship of APP-mediated neurotoxicity to JNK is not clear, partly because APP cytotoxicity is Abeta independent. Here we examined whether JNK is involved in APP-mediated neuronal cell death and found that: (i) neuronal cell death by antibody-bound APP was inhibited by dominant-negative JNK, JIP-1b and SP600125, the specific inhibitor of JNK, but not by SB203580 or PD98059; (ii) constitutively active (ca) JNK caused neuronal cell death and (iii) the pharmacological profile of caJNK mediated cell death closely coincided with that of APP-mediated cell death. Pertussis toxin (PTX) suppressed APP-mediated cell death but not caJNK-induced cell death, which was suppressed by Humanin, a newly identified neuroprotective factor which inhibits APP-mediated cytotoxicity. In the presence of PTX, the PTX resistant mutant of Galphao, but not that of Galphai, recovered the cytotoxic action of APP. These findings demonstrate that JNK is involved in APP-mediated neuronal cell death as a downstream signal transducer of Go. PMID- 12562531 TI - Methyl-beta-cyclodextrin but not retinoic acid reduces EAAT3-mediated glutamate uptake and increases GTRAP3-18 expression. AB - The Na+-dependent glutamate transporter EAAT3 facilitates glutamate uptake into neurons as well as many other cell types. GTRAP3-18 (JWA, Arl6ip5) is a novel protein that interacts with EAAT3 and negatively modulates EAAT3-mediated glutamate uptake. Previous studies suggest that retinoic acid (RA) decreases Na+ dependent glutamate uptake and increases GTRAP3-18 protein expression. However, the RA used in those studies was complexed with methyl-beta-cyclodextrin (MebetaCD). In the present study we found that MebetaCD, but not RA, significantly reduced Na+-dependent EAAT3-mediated [3H]glutamate uptake in human embryonic kidney 293 (HEK293) cells. MebetaCD also significantly increased GTRAP3 18 protein expression in HEK293 cells as well as in rat hypothalamic neuron cultures. Intracerebroventricular administration of MebetaCD to the mouse brain resulted in a significant increase in GTRAP3-18 immunoreactivity in the hippocampus and cerebral cortex. In conclusion, we have shown that MebetaCD reduces EAAT3-mediated glutamate uptake and induces the expression of GTRAP3-18 protein. PMID- 12562532 TI - Risk prediction: increasingly important, but questions still remain. PMID- 12562533 TI - C-reactive protein and lipids may respond differently to statins. PMID- 12562534 TI - Issues that led to the withdrawal of cerivastatin. PMID- 12562535 TI - Patient compliance is a critical factor in the effectiveness of cardiac prevention. PMID- 12562536 TI - The challenges of conducting clinical endpoint studies. AB - In order to evaluate the effects of a particular treatment strategy on mortality and major morbidity within a disease entity, large, multinational, relatively long-term clinical endpoint studies are often conducted. The primary challenge of conducting these studies is to maintain consistency in the interpretation of the clinical endpoints across different geographic areas and over the long time course of the study. The success of a clinical endpoint study depends on understanding the challenges and incorporating the special requirements of these studies into the protocol design and operational procedures throughout the study. PMID- 12562537 TI - Efficacy of over-the-counter nutritional supplements. AB - More than 100 million people in the United States report using nutritional supplements. Most people are under the impression that nutritional supplements offer health benefits and are closely regulated to ensure safety and efficacy. Unfortunately, the Dietary Supplement Health and Education Act of 1994 allows for the promotion of nutritional supplements without review by the United States Food and Drug Administration; therefore, it is important to evaluate the efficacy and safety of these supplements. There is strong scientific evidence supporting the use of plant sterols/stanols, omega-3 fatty acids, niacin, folate, vitamin B(6)/B(12), and tree nuts. There is potential evidence for the health benefits of soy protein, tea extracts, policosanol, guggulipids, coenzyme Q10, and L arginine. There has been a lack of evidence for the health benefits of garlic and antioxidants. PMID- 12562538 TI - Small, dense low-density lipoprotein: risk or myth? AB - Based on the particle diameter of the major subpopulation of low-density lipoprotein (LDL) in plasma, an individual may be classified either as having phenotype A (desirable phenotype; large, buoyant LDL) or phenotype B (high risk; small, dense LDL). This article reviews the clinical significance of LDL particle diameter determination and proposes a strategy for incorporating this information in the new guidelines of the National Cholesterol Education Program's Adult Treatment Panel III. PMID- 12562539 TI - The development of combination drugs for atherosclerosis. AB - The complexity of medical care has led to a dramatic increase in the number of drugs taken by patients. In an effort to improve patient compliance and the effectiveness of clinical care, many drugs are being studied for combination in a single capsule or tablet. The recent success of many combination drugs for hypertension has accelerated the interest in combination drugs for lipids and atherosclerosis. Advicor (Kos Pharmaceuticals, Miami, FL), a combination of nicotinic acid and a statin (mevacor), is the first combination lipid drug. Recently, the United States Food and Drug Administration approved the combination of another statin, pravachol, with aspirin. Therefore, combination drugs are likely to be a significant contribution to clinical practice and drug development. PMID- 12562540 TI - Vascular pleiotropy of statins: clinical evidence and biochemical mechanisms. AB - The ability of statins to lower serum cholesterol and reduce coronary heart disease endpoints has confirmed portions of the lipid hypothesis. However, the time to benefit and increased benefit in overlapping populations have suggested that nonlipid or pleiotropic effects of statins may be present. The apparent benefit of statins in cerebrovascular disease may imply a similar final common pathway among the diverse mechanisms of vascular diseases. Statins' inhibition of isoprenoid intermediates may modify GTP binding proteins such as Rho. The augmentation of collateral blood flow downstream of activated plaque through endothelial cell nitric oxide synthase may be the biochemical basis of statins' vascular pleiotropy. Eventual clinical paradigms of statin use may include higher doses to enhance pleiotropic effects and treatment, even when lipid markers are within guidelines. PMID- 12562541 TI - Are cholesterol-lowering medications and antihypertensive agents preventing stroke in ways other than by controlling the risk factor? AB - Statins and angiotensin-converting enzyme (ACE) inhibitors are an important component of our armamentarium for stroke prevention. Both of these classes of agents have a primary mechanism of action of reducing the level of the respective risk factor. They also have mechanisms of action that may confer benefits beyond what is believed to be the primary action of the agent. This has led to speculation that statins reduce stroke risk by means beyond cholesterol lowering, and ACE inhibitors reduce stroke risk by means beyond blood pressure lowering. We review the mounting evidence that suggests that statins and ACE inhibitors have so-called pleiotropic effects that may lead to stroke prevention. PMID- 12562542 TI - Early use of statins in acute coronary syndromes. AB - This review examines the use of statin medications early in the clinical course of acute coronary syndrome (ACS). Available data demonstrate that there are clear clinical benefits to this practice. Numerous previous studies have documented the primary and secondary benefits of statins in the prevention of coronary events. Recent trials show that when statins are used during hospital admissions for ACS, patients experience decreased recurrent myocardial infarction, lower death rates, and fewer repeat hospitalizations for ischemia or revascularization. Several studies suggest that the positive effects of statins on plaque stabilization, inflammation, thrombosis, and endothelial function may be independent of lipid levels. There is also an emerging view that beneficial lipid-lowering with statins in high-risk patients has no lower limit. This information suggests that all patients admitted for ACS should be treated with statins, regardless of cholesterol levels. PMID- 12562543 TI - Naturally available oils contain phytosterols that affect cholesterol absorption. PMID- 12562544 TI - What is the cardioprotective role of hormone replacement therapy? AB - A very large body of literature has yielded strong biologic and mechanistic plausibility for the consistent observational findings that estrogen is cardioprotective. Recently completed randomized, controlled trials have been interpreted as challenging the doctrine that hormone replacement is cardioprotective for postmenopausal women. However, other than the Estrogen in the Prevention of Atherosclerosis Trial, none of the currently completed (and no ongoing) randomized, controlled trials have appropriately tested the hypothesis generated from observational data that estrogen replacement is cardioprotective. This mainly results from the fact that randomized, controlled trials have not tested the same pattern and type of hormone use in the same population of women observed in the epidemiologic studies. On the other hand, recently completed randomized, controlled trials provide important but limited information concerning the clinical use of a specific regimen of hormone replacement for the prevention of cardiovascular disease in a particular population of postmenopausal women. Observations made from epidemiologic studies will have to be appropriately tested in randomized, controlled trials before any real conclusions can be drawn as to whether hormone replacement is cardioprotective. PMID- 12562545 TI - Pharmacoeconomics of lipid-lowering drugs. AB - The widespread use and high cost of lipid-lowering drugs have resulted in numerous investigations of their cost effectiveness. Pharmacoeconomic analyses have considered initiation of treatment, maintenance therapy, and long-term outcomes of lipid-lowering therapy. Analyses have yielded a number of consistent messages, but the story continues to evolve as new trials, models, and drugs are introduced. Certain medications are more efficacious and associated with more cost-effective lipid-lowering therapy initiation. Other medications are lower priced and may be more cost effective for maintenance, contingent upon having achieved lipid-lowering goals. Finally, some medications have proven long-term cost effectiveness in trials and comparative cost effectiveness in models. Research is certain to further our understanding of the costs of lipid-lowering therapy and the methods needed to optimize expenditures. PMID- 12562546 TI - Cardiovascular and medical ramifications of treatment of subclinical hypothyroidism. AB - Subclinical hypothyroidism can be diagnosed in 1% to 10% of the adult population, is more common in women, and increases with age. In many patients, treatment with L-thyroxine reduces low-density lipoprotein cholesterol, improves cardiac function, reduces symptoms of hypothyroidism, and diminishes neuropsychiatric symptoms. Treatment also reduces the likelihood of statin-induced myopathy. However, in double-blind, placebo-controlled trials of L-thyroxine therapy in subclinical hypothyroidism, cardiovascular and symptomatic benefits have been neither uniform nor definitive. In the absence of a large-scale, multicenter, randomized trial, physicians have to individualize therapy for each patient. Benefits of therapy are most likely to be realized in patients with thyroid stimulating hormone levels greater than 10 mU/L on repeated measures, those with hypothyroid symptoms, those who are pregnant, those with a documented family history of hypothyroidism, and those with severe hyperlipidemia. PMID- 12562548 TI - Interaction of environmental allergens with airway epithelium as a key component of asthma. AB - Epithelial cells in the airway wall actively interact with environmental antigens/allergens, both in healthy individuals and patients with asthma. In patients with (allergic) asthma, the epithelium is abnormal, showing damaged structures and continuous activation similar to a repair phenotype cell. Epithelial cells bind allergens by a diversity of innate receptors, similar and in part identical to the Toll-like receptor family, which can induce the release of cytokines, chemokines, and growth factors. Protease-containing extracts (house dust mite, fungi) may additionally cause damage of the epithelial cell layer, thereby enhancing the repair phenotype of epithelial cells in patients with asthma. These interactions may result in facilitation of transport of allergens and enhanced presentation to the immune system (Th2-type response). The inflammatory response induces a second phase of Th2-type cytokines and cytotoxic products that will enhance growth factor-mediated airway remodeling, as is found in asthma. An understanding of the largely unknown innate responses of epithelial cells with environmental antigens/allergens may open new treatment modalities for asthma and other airway diseases. PMID- 12562549 TI - Childhood asthma. AB - Asthma is the most common chronic disease of childhood. Despite increasing evidence indicating that genetic and environmental factors have significant effects on airway development and function in early life, our understanding of the natural history of the disease is limited. The development of objective outcome measures will enable us to distinguish between children with transient early wheeze and those who will progress to persistent asthma, and to investigate factors affecting airway function throughout early childhood. A thorough knowledge of lung physiology in early life and its relationship with environmental exposures and prior or subsequent development of asthma would have a profound effect on our understanding of the natural history of asthma and the design of future strategies for disease prevention. PMID- 12562550 TI - Indoor allergen exposure in the development of allergy and asthma. AB - Research evidence supports a causative relationship between indoor allergen exposure and the development of asthma. Epidemiologic studies demonstrate that exposure to indoor allergens, particularly house-dust mites and cockroaches and, to a lesser extent, animal allergens and mold, is a risk factor for the development of sensitization and perhaps respiratory symptoms. Sensitization to indoor allergen is clearly a major risk factor for the development of asthma and allergy. There is also epidemiologic and experimental evidence that, in sensitized subjects, higher exposure to indoor allergen causes morbidity. Most, but not all, prospective studies support the causative link between allergen exposure and asthma. Additional evidence comes from the prevention studies, in which reduction in indoor allergen exposure may lead to improvement in symptoms, pulmonary function, and bronchial hyperresponsiveness, and, if practiced, in early years of life, perhaps the development of asthma and allergy. PMID- 12562552 TI - Dietary factors in the pathogenesis of asthma and chronic obstructive pulmonary disease. AB - This review focuses on the role of dietary factors in the pathogenesis of asthma and chronic obstructive pulmonary disease. Reduced pulmonary function and indicators of airway obstruction have been shown to be strong indicators of mortality; therefore, a link with diet in the pathogenesis of these diseases would have important public health implications. The results of the studies identified in the review suggest that people who have a diet rich in fruit and vegetables have a lower risk of poor respiratory health, and that this may be due to the antioxidant nutrients these foods contain. On the basis of the evidence, it seems justified to promote a healthy diet, high in fruits, vegetables, and whole grain foods and low in alcohol and fatty foods, as set out in existing guidelines for prevention of cardiovascular disease and cancer, to protect respiratory health in both children and adults. PMID- 12562551 TI - Nitric oxide, nitrotyrosine, and nitric oxide modulators in asthma and chronic obstructive pulmonary disease. AB - Nitric oxide (NO), a simple free-radical gas, elicits a diverse range of physiologic and pathophysiologic effects, and plays an important role in pulmonary diseases. Nitrosative stress and nitration of proteins in airway epithelium may be responsible for steroid resistance in asthma and their ineffectiveness in chronic obstructive pulmonary disease (COPD), supporting the potential role of future therapeutic strategies aimed at regulating NO synthesis in asthma and COPD. In this article, we review the potential role of NO modulators (NO synthase inhibitors and NO donors), which, if given on a regular basis, may have clinical benefit in asthma and COPD. PMID- 12562553 TI - Rhinoviruses in the pathogenesis of asthma. AB - Using sensitive diagnostic methodologies, epidemiologic studies during the past decade have allowed the identification of human rhinoviruses (RVs), generally recognized as "common cold viruses," as major asthma precipitants. This association was further established by evaluating the impact of RV infection in airway obstruction and inflammation during naturally acquired or experimentally induced RV colds. There is now strong evidence that RVs can infect and propagate not only in the upper but also in the lower airways. Bronchial and pulmonary epithelia infected by RVs are rich sources of inflammatory mediators, which may initiate or augment airway inflammation and obstruction. Furthermore, in an atopic environment, responses to the virus are skewed by and toward an "atopic," Th2-like balance, which may further enhance inflammation and exacerbate asthma. PMID- 12562554 TI - Diesel fumes and the rising prevalence of atopy: an urban legend? AB - Recently, the incidence of allergic diseases has increased in most industrialized countries of the world. Persistent exposure to particulate air pollution from motor vehicles has been implicated as one of the factors that is responsible for the observed increased prevalence of atopy. Epidemiologic studies conducted in different parts of the world have demonstrated an important association between ambient levels of motor vehicle traffic emissions and increased symptoms of asthma and rhinitis. Additionally, recent human and animal laboratory-based studies have shown that particulate toxic pollutants, and in particular diesel exhaust particles (DEP), can enhance allergic inflammation and induce the development of allergic immune responses. In this article, our current understanding of the mechanisms by which pollutants such as DEPs enhance the underlying allergic inflammatory response is reviewed, and the evidence that supports the causative link between particulate air pollution from motor vehicles and increasing allergic diseases is discussed. PMID- 12562555 TI - Airway vascular remodeling in asthma. AB - Several characteristic changes occur in the bronchial wall in asthma, including specific changes to the vasculature. These result in an increase in vessel numbers per unit area, as well as increased vessel activity suggested by vasodilatation, vessel leakage, and cellular margination with transmigration to target tissues. This combined action in asthma leads to airway-wall thickening and reduced airflow. Each component of the vascular response has been shown to be controlled by a range of inflammatory mediators and growth factors. These factors are themselves regulated by a complex process initially involving gene expression, transcription, and translation at the molecular level, then subsequent protein release, binding to matrix elements, endothelial cell activation, and a proliferative endothelial response. Many commonly used airway medications are capable of modulating the vascular response to inflammatory stimuli. New therapies might improve airflow through better regulation of vessel growth, dilatation, and leakage in the airway wall. PMID- 12562556 TI - Neutrophils in asthma pathophysiology. AB - Although the role of eosinophils, mast cells, and T cells in asthma has long been recognized, several reports suggest that neutrophils may also be involved. In most studies of people with mild asthma, neutrophil numbers in the airways are not different from controls. However, in severe asthma, including asthma deaths, neutrophils are usually raised. Furthermore, most pediatric studies suggest that neutrophils are raised in some children, in particular in the young or infantile wheeze group. Measurements of inflammatory mediators in the airways of asthmatic subjects suggest that neutrophils are activated. PMID- 12562557 TI - Physiopathology of airway hyperresponsiveness. AB - Airway hyperresponsiveness (AHR), the tendency of the airways to narrow too much and too easily in response to various stimuli, is a universal feature of asthma, although it is not exclusive to this disease. Airway responsiveness shows a unimodal distribution in the general population and might vary with time, increasing after exposure to allergens, industrial substances, or infectious agents in predisposed individuals, or decreasing for variable time periods after environmental or pharmacologic interventions. Airway inflammation and structural airway changes can lead to this heightened airway response, but the mechanisms by which they modify airway function are still unclear. They might be associated with increased contractile properties of the airways--from an increase in contractile elements, a change in smooth muscle mechanical properties, or a reduction of forces opposing bronchoconstriction, such as reduced airway parenchymal interdependence. Other factors, such as neurohumoral influences and "geometric factors" (eg, airway caliber), can modulate the degree of AHR. PMID- 12562558 TI - Single-isomer levalbuterol: a review of the acute data. AB - Levalbuterol, the pure (R)-isomer of racemic albuterol, is a new therapeutic option for patients with asthma. Racemic albuterol comprises a 50:50 mixture of (R)- and (S)-albuterol, with (R)-albuterol conferring all of the bronchodilator effects of the racemate. Numerous preclinical and in vitro studies have indicated that (S)-albuterol is not an inert isomer, but may have proinflammatory effects. Results from clinical trials in adults and children with asthma have demonstrated that 0.63 mg levalbuterol provides effective bronchodilation with lower b mediated side effects compared with 2.5 mg racemic albuterol. In the emergency department, levalbuterol provided greater bronchodilation and significantly reduced hospital admissions compared with racemic albuterol. Recent studies have supported that levalbuterol use in acute settings may reduce the cost of asthma treatment by decreasing the total treatments and subsequent respiratory therapy resources. Levalbuterol provides heath care professionals with a safe, effective, and potentially cost-saving alternative to racemic albuterol for the treatment of patients with asthma. PMID- 12562559 TI - Managing outpatient asthma exacerbations. AB - Asthma is a chronic inflammatory disease that renders individuals prone to acute exacerbations. Several allergic and nonallergic triggers can incite an asthma exacerbation. The goals of managing an asthma exacerbation are prompt recognition, rapid reversal of airflow obstruction, prevention of relapses, and forestalling future episodes. A written asthma home-management plan is essential to minimize the severity of exacerbations. Short-acting b-agonists, oxygen, and corticosteroids are fundamental to early intervention in acute asthma exacerbation. Anticholinergics and magnesium sulfate can help nonresponders. Newer agents such as levalbuterol and long-acting b-agonists might be future additions to our armamentarium of drugs to treat acute exacerbations. Initiation or intensification of long-term controller therapy, treatment of co-morbid conditions, and avoidance of possible triggers along with prompt follow-up can help prevent relapses. Listening to patient preferences and concerns to enhance adherence and regular follow-up care can help prevent future episodes. PMID- 12562560 TI - ChTX induces oscillatory contraction in guinea pig trachea: role of cyclooxygenase-2 and PGE2. AB - We examined the possible role of cyclooxygenase (COX) in charybdotoxin (ChTX) induced oscillatory contraction in guinea pig trachea. Involvement of prostaglandin E(2) (PGE(2)) in ChTX-induced oscillatory contraction was also investigated. ChTX (100 nM) induced oscillatory contraction in guinea pig trachea. The mean oscillatory frequency induced by ChTX was 10.7 +/- 0.8 counts/h. Maximum and minimum tensions within ChTX-induced oscillatory contractions were 68.4 +/- 1.8 and 14.3 +/- 1.7% compared with K(+) (72.7 mM) contractions. ChTX-induced oscillatory contraction was completely inhibited by indomethacin, a nonselective COX inhibitor. Valeryl salicylate, a selective COX-1 inhibitor, did not significantly inhibit this contraction, whereas N-(2 cyclohexyloxy-4-nitro-phenyl)-methanesulfonamide, a selective COX-2 inhibitor, abolished this contraction. Exogenously applied arachidonic acid enhanced ChTX induced oscillatory contraction. SC-51322, a selective PGE receptor subtype EP(1) antagonist, significantly inhibited ChTX-induced oscillatory contraction. Exogenously applied PGE(2) induced only a slight phasic contraction in guinea pig trachea, but PGE(2) induced strong oscillatory contraction after pretreatment with indomethacin and ChTX. Moreover, ChTX time-dependently stimulated PGE(2) generation. These results suggest that ChTX specifically activates COX-2 and stimulates PGE(2) production and that ChTX-induced oscillatory contraction in guinea pig trachea is mediated by activation of EP(1) receptor. PMID- 12562561 TI - Classical isoforms of PKC as regulators of CAT-1 transporter activity in pulmonary artery endothelial cells. AB - We examined which isoforms of protein kinase C (PKC) may be involved in the regulation of cationic amino acid transporter-1 (CAT-1) transport activity in cultured pulmonary artery endothelial cells (PAEC). An activator of classical and novel isoforms of PKC, phorbol 12-myristate-13-acetate (PMA; 100 nM), inhibited CAT-1-mediated l-arginine transport in PAEC after a 1-h treatment and activated l arginine uptake after an 18-h treatment of cells. These changes in l-arginine transport were not related to the changes in the expression of the CAT-1 transporter. The inhibitory effect of PMA on l-arginine transport was accompanied by a translocation of PKCalpha (a classical PKC isoform) from the cytosol to the membrane fraction, whereas the activating effect of PMA on l-arginine transport was accompanied by full depletion of the expression of PKCalpha in PAEC. A selective activator of Ca(2+)-dependent classical isoforms of PKC, thymeleatoxin (Thy; 100 nM; 1-h and 18-h treatments), induced the same changes in l-arginine uptake and PKCalpha translocation and depletion as PMA. The effects of PMA and Thy on l-arginine transport in PAEC were attenuated by a selective inhibitor of classical PKC isoforms Go 6976 (1 micro M). Phosphatidylinositol-3,4,5 triphosphate-dipalmitoyl (PIP; 5 micro M), which activates novel PKC isoforms, did not affect l-arginine transport in PAEC after 1-h and 18-h treatment of cells. PIP (5 micro M; 1 h) induced the translocation of PKCepsilon (a novel PKC isoform) from the cytosolic to the particulate fraction and did not affect the translocation of PKCalpha. These results demonstrate that classical isoforms of PKC are involved in the regulation of CAT-1 transport activity in PAEC. We suggest that translocation of PKCalpha to the plasma membrane induces phosphorylation of the CAT-1 transporter, which leads to inhibition of its transport activity in PAEC. In contrast, depletion of PKCalpha after long-term treatment with PMA or Thy promotes dephosphorylation of the CAT-1 transporter and activation of its activity. PMID- 12562562 TI - High tidal volume ventilation induces NOS2 and impairs cAMP- dependent air space fluid clearance. AB - Tidal volume reduction during mechanical ventilation reduces mortality in patients with acute lung injury and the acute respiratory distress syndrome. To determine the mechanisms underlying the protective effect of low tidal volume ventilation, we studied the time course and reversibility of ventilator-induced changes in permeability and distal air space edema fluid clearance in a rat model of ventilator-induced lung injury. Anesthetized rats were ventilated with a high tidal volume (30 ml/kg) or with a high tidal volume followed by ventilation with a low tidal volume of 6 ml/kg. Endothelial and epithelial protein permeability were significantly increased after high tidal volume ventilation but returned to baseline levels when tidal volume was reduced. The basal distal air space fluid clearance (AFC) rate decreased by 43% (P < 0.05) after 1 h of high tidal volume but returned to the preventilation rate 2 h after tidal volume was reduced. Not all of the effects of high tidal volume ventilation were reversible. The cAMP dependent AFC rate after 1 h of 30 ml/kg ventilation was significantly reduced and was not restored when tidal volume was reduced. High tidal volume ventilation also increased lung inducible nitric oxide synthase (NOS2) expression and air space total nitrite at 3 h. Inhibition of NOS2 activity preserved cAMP-dependent AFC. Because air space edema fluid inactivates surfactant and reduces ventilated lung volume, the reduction of cAMP-dependent AFC by reactive nitrogen species may be an important mechanism of clinical ventilator-associated lung injury. PMID- 12562563 TI - From Giacometti to Botero: images of eating disorders under investigation. PMID- 12562564 TI - Gene therapy for psychiatric disorders. AB - There has been tremendous progress in developing techniques for manipulating genetic material, and the birth of gene therapy as a discipline has been one consequence of this. Most considerations of gene therapy in the nervous system have focused on attempts to transfer novel genes in for the purpose of protecting neurons from neurological insults. In this review, the author considers the progress in that field and the possible application of related gene therapy approaches to the far more difficult task of buffering against a psychiatric disorder. As an emphasis, the author reviews how the biology of psychiatric disorders is so often one of vulnerability to particular environments. Because of this context dependency, it would be likely that many possible gene therapeutic interventions would need to be context dependent as well. Thus, the author considers the plausibility of developing gene vector therapies that use conditional expression systems, in particular ones whose expression would be induced by the same environmental perturbations that exacerbate psychiatric symptoms themselves. In particular, the author considers the role of stress as a predisposing factor in certain psychiatric disorders and the ways in which stress signals can be harnessed as inducers of conditional expression systems in gene therapy. PMID- 12562565 TI - Chronic fatigue syndrome: a review. AB - OBJECTIVE: Chronic fatigue syndrome is an illness characterized by disabling fatigue of at least 6 months, accompanied by several other symptoms. This review summarizes the current state of knowledge about chronic fatigue syndrome. METHOD: The case definition, prevalence, clinical presentation, evaluation, and prognosis of chronic fatigue syndrome are discussed. Research on the pathophysiology and treatment of chronic fatigue syndrome is reviewed. RESULTS: Chronic fatigue syndrome is diagnosed on the basis of symptoms. Patients with chronic fatigue syndrome experience significant functional impairment. Pathophysiological abnormalities exist across many domains, suggesting that chronic fatigue syndrome is a heterogeneous condition of complex and multifactorial etiology. Evidence also is beginning to emerge that chronic fatigue syndrome may be familial. Although chronic fatigue syndrome has significant symptom overlap and comorbidity with psychiatric disorders, several lines of research suggest that the illness may be distinct from psychiatric disorders. Patients' perceptions, attributions, and coping skills, however, may help perpetuate the illness. Treatment for chronic fatigue syndrome is symptom-based and includes pharmacological and behavioral strategies. Cognitive behavior therapy and graded exercise can be effective in treating the fatigue and associated symptoms and disability. CONCLUSIONS: Chronic fatigue syndrome is unlikely to be caused or maintained by a single agent. Findings to date suggest that physiological and psychological factors work together to predispose an individual to the illness and to precipitate and perpetuate the illness. The assessment and treatment of chronic fatigue syndrome should be multidimensional and tailored to the needs of the individual patient. PMID- 12562566 TI - Depression, IV: STAR*D treatment trial for depression. PMID- 12562569 TI - Childhood obsessive-compulsive personality traits in adult women with eating disorders: defining a broader eating disorder phenotype. AB - OBJECTIVE: The authors retrospectively examined a spectrum of childhood traits that reflect obsessive-compulsive personality in adult women with eating disorders and assessed the predictive value of the traits for the development of eating disorders. METHOD: In a case-control design, 44 women with anorexia nervosa, 28 women with bulimia nervosa, and 28 healthy female comparison subjects were assessed with an interview instrument that asked them to recall whether they had experienced various types of childhood behavior suggesting traits associated with obsessive-compulsive personality. The subjects also completed a self-report inventory of obsessive-compulsive disorder (OCD) symptoms. RESULTS: Childhood obsessive-compulsive personality traits showed a high predictive value for development of eating disorders, with the estimated odds ratio for eating disorders increasing by a factor of 6.9 for every additional trait present. Subjects with eating disorders who reported perfectionism and rigidity in childhood had significantly higher rates of obsessive-compulsive personality disorder and OCD comorbidity later in life, compared with eating disorder subjects who did not report those traits. CONCLUSIONS: Childhood traits reflecting obsessive-compulsive personality appear to be important risk factors for the development of eating disorders and may represent markers of a broader phenotype for a specific subgroup of patients with anorexia nervosa. PMID- 12562570 TI - Risk factors for the onset of eating disorders in adolescent girls: results of the McKnight longitudinal risk factor study. AB - OBJECTIVE: This study examined the importance of potential risk factors for eating disorder onset in a large multiethnic sample followed for up to 3 years, with assessment instruments validated for the target population and a structured clinical interview used to make diagnoses. METHOD: Participants were 1,103 girls initially assessed in grades 6-9 in school districts in Arizona and California. Each year, students completed the McKnight Risk Factor Survey, had body height and weight measured, and underwent a structured clinical interview. The McKnight Risk Factor Survey, a self-report instrument developed for this age group, includes questions related to risk factors for eating disorders. RESULTS: During follow-up, 32 girls (2.9%) developed a partial- or full-syndrome eating disorder. At the Arizona site, there was a significant interaction between Hispanics and higher scores on a factor measuring thin body preoccupation and social pressure in predicting onset of eating disorders. An increase in negative life events also predicted onset of eating disorders in this sample. At the California site, only thin body preoccupation and social pressure predicted onset of eating disorders. A four-item screen derived from thin body preoccupation and social pressure had a sensitivity of 0.72, a specificity of 0.80, and an efficiency of 0.79. CONCLUSIONS: Thin body preoccupation and social pressure are important risk factors for the development of eating disorders in adolescents. Some Hispanic groups are at risk of developing eating disorders. Efforts to reduce peer, cultural, and other sources of thin body preoccupation may be necessary to prevent eating disorders. PMID- 12562571 TI - Topiramate in the treatment of binge eating disorder associated with obesity: a randomized, placebo-controlled trial. AB - OBJECTIVE: Binge eating disorder is associated with obesity. Topiramate is an antiepileptic agent associated with weight loss. The objective of this study was to evaluate topiramate in the treatment of binge eating disorder associated with obesity. METHOD: For this 14-week, double-blind, flexible-dose (25-600 mg/day) topiramate trial, 61 outpatients (53 women, eight men) with binge eating disorder who were obese (body mass index >/=30 kg/m(2)) were randomly assigned to receive topiramate (N=30) or placebo (N=31). The primary efficacy measure was binge frequency. The primary analysis of efficacy was a repeated-measures random regression with treatment-by-time as the effect measure. RESULTS: Compared with placebo, topiramate was associated with a significantly greater rate of reduction in binge frequency, binge day frequency, body mass index, weight, and scores on the Clinical Global Impression severity scale and the Yale-Brown Obsessive Compulsive Scale (modified for binge eating). Topiramate was also associated with significantly greater reductions in binge frequency (topiramate: 94%, placebo: 46%) and binge day frequency (topiramate: 93%, placebo: 46%) and with a significantly higher level of response than placebo. The mean weight loss for topiramate-treated subjects who completed the study was 5.9 kg. Median topiramate dose was 212 mg/day (range=50-600). Nine patients (three receiving placebo, six given topiramate) discontinued because of adverse events. The most common reasons for discontinuing topiramate were headache (N=3) and paresthesias (N=2). CONCLUSIONS: Topiramate was efficacious and relatively well tolerated in the short-term treatment of binge eating disorder associated with obesity. PMID- 12562572 TI - Differential effects of developmental cerebellar abnormality on cognitive and motor functions in the cerebellum: an fMRI study of autism. AB - OBJECTIVE: Recent years have seen a revolution in views regarding cerebellar function. New findings suggest that the cerebellum plays a role in multiple functional domains: cognitive, affective, and sensory as well as motor. These findings imply that developmental cerebellar pathology could play a role in certain nonmotor functional deficits, thereby calling for a broader investigation of the functional consequences of cerebellar pathology. Autism provides a useful model, since over 90% of autistic cerebella examined at autopsy have shown well defined cerebellar anatomic abnormalities. The aim of the present study was to examine how such pathology ultimately impacts cognitive and motor function within the cerebellum. METHOD: Patterns of functional magnetic resonance imaging (fMRI) activation within anatomically defined cerebellar regions of interest were examined in eight autistic patients (ages 14-38 years) and eight matched healthy comparison subjects performing motor and attention tasks. For the motor task, subjects pressed a button at a comfortable pace, and activation was compared with a rest condition. For the attention task, visual stimuli were presented one at a time at fixation, and subjects pressed a button to every target. Activation was compared with passive visual stimulation. RESULTS: While performing these tasks, autistic individuals showed significantly greater cerebellar motor activation and significantly less cerebellar attention activation. CONCLUSIONS: These findings shed new light on the cerebellar role in attention deficits in autism and suggest that developmental cerebellar abnormality has differential functional implications for cognitive and motor systems. PMID- 12562573 TI - The longitudinal course of borderline psychopathology: 6-year prospective follow up of the phenomenology of borderline personality disorder. AB - OBJECTIVE: The syndromal and subsyndromal phenomenology of borderline personality disorder was tracked over 6 years of prospective follow-up. METHOD: The psychopathology of 362 inpatients with personality disorders was assessed with the Revised Diagnostic Interview for Borderlines (DIB-R) and borderline personality disorder module of the Revised Diagnostic Interview for DSM-III-R Personality Disorders. Of these patients, 290 met DIB-R and DSM-III-R criteria for borderline personality disorder and 72 met DSM-III-R criteria for other axis II disorders (and neither criteria set for borderline personality disorder). Most of the borderline patients received multiple treatments before the index admission and during the study. Over 94% of the total surviving subjects were reassessed at 2, 4, and 6 years by interviewers blind to previously collected information. RESULTS: Of the subjects with borderline personality disorder, 34.5% met the criteria for remission at 2 years, 49.4% at 4 years, 68.6% at 6 years, and 73.5% over the entire follow-up. Only 5.9% of those with remissions experienced recurrences. None of the comparison subjects with other axis II disorders developed borderline personality disorder during follow-up. The patients with borderline personality disorder had declining rates of 24 symptom patterns but remained symptomatically distinct from the comparison subjects. Impulsive symptoms resolved the most quickly, affective symptoms were the most chronic, and cognitive and interpersonal symptoms were intermediate. CONCLUSIONS: These results suggest that symptomatic improvement is both common and stable, even among the most disturbed borderline patients, and that the symptomatic prognosis for most, but not all, severely ill borderline patients is better than previously recognized. PMID- 12562574 TI - Impaired fasting glucose tolerance in first-episode, drug-naive patients with schizophrenia. AB - OBJECTIVE: This study examined the prevalence of impaired fasting glucose tolerance in first-episode, drug-naive patients with schizophrenia. METHOD: In this cross-sectional study, fasting plasma levels of glucose, insulin, lipids, and cortisol were measured in 15 male and 11 female hospitalized Caucasian patients with DSM-IV schizophrenia (mean age=33.6 years) and age- and sex-matched healthy comparison subjects. The patients and comparison subjects were also matched in terms of various life-style and anthropometric measures. RESULTS: More than 15% of the drug-naive, first-episode patients with schizophrenia had impaired fasting glucose tolerance, compared to none of the healthy volunteers. Compared with the healthy subjects, the patients with schizophrenia had significantly higher fasting plasma levels of glucose (mean=88.2 mg/dl, SD=5.4, for the healthy subjects versus mean=95.8 mg/dl, SD=16.9, for the patients), insulin (mean=7.7 micro u/ml, SD=3.7, versus mean=9.8 micro u/ml, SD=3.9), and cortisol (mean=303.2 nmol/liter, SD=10.5, versus mean=499.4 nmol/liter, SD=161.4) and were more insulin resistant, as measured with homeostasis model assessment (mean=1.7, SD=0.7, for the healthy subjects versus mean=2.3, SD=1.0, for the patients). CONCLUSIONS: First-episode, drug-naive patients with schizophrenia have impaired fasting glucose tolerance and are more insulin resistant and have higher levels of plasma glucose, insulin, and cortisol than healthy comparison subjects. PMID- 12562576 TI - Attenuation of olanzapine-induced weight gain with reboxetine in patients with schizophrenia: a double-blind, placebo-controlled study. AB - OBJECTIVE: Since increased norepinephrine availability may account for the weight reducing effect of appetite suppressants, the authors hypothesized that the addition of the selective norepinephrine reuptake inhibitor reboxetine may prevent or attenuate olanzapine-induced weight gain. METHOD: Twenty-six patients hospitalized for first-episode DSM-IV schizophrenic disorder participated in the study. In addition to 6 weeks of treatment with olanzapine, 10 mg/day, patients were randomly allocated in a double-blind design to receive either reboxetine, 4 mg/day, (N=13) or placebo (N=13). RESULTS: Ten patients in each group completed the 6-week trial. Patients given olanzapine and reboxetine demonstrated a significantly lower increase in body weight (mean=2.5 kg, SD=2.7) than those given olanzapine and placebo (mean=5.5 kg, SD=3.1). Significantly fewer patients in the olanzapine/reboxetine group (N=2 of 10) than in the olanzapine/placebo group (N=7 of 10) gained at least 7% of their initial weight, the cutoff for clinically significant weight gain. The addition of reboxetine to olanzapine treatment was safe and well tolerated by the patients. A between-group difference in the reduction of Hamilton depression scale scores was seen that favored the olanzapine/reboxetine group (mean difference=-3.1, SD=1.25). CONCLUSIONS: The selective norepinephrine reuptake inhibitor reboxetine may reduce olanzapine induced weight gain in schizophrenia patients, and activation of the adrenergic system may attenuate weight gain induced by atypical antipsychotic agents. PMID- 12562575 TI - Changes in glucose and cholesterol levels in patients with schizophrenia treated with typical or atypical antipsychotics. AB - OBJECTIVE: The association of hyperglycemia and hypercholesterolemia with use of atypical antipsychotics has been documented in case reports and uncontrolled studies. The authors' goal was to assess the effects of clozapine, olanzapine, risperidone, and haloperidol on glucose and cholesterol levels in hospitalized patients with schizophrenia or schizoaffective disorder during a randomized double-blind 14-week trial. METHOD: One hundred fifty-seven patients with schizophrenia or schizoaffective disorder who were inpatients at four hospitals were originally included in the study. The 14-week trial consisted of an 8-week fixed-dose period and a 6-week variable-dose period. Planned assessments included fasting glucose and cholesterol, which were collected at baseline and at the end of the 8-week period and the following 6-week period. RESULTS: One hundred eight of the 157 patients provided blood samples at baseline and at least at one point after random assignment to clozapine, olanzapine, risperidone, or haloperidol during the treatment trial. Seven of these patients had diabetes; their glucose levels were >125 mg/dl at baseline. Data from 101 patients were used for statistical analyses. During the initial 8-week period there was an overall significant increase in mean glucose levels. There were significant increases in glucose levels at the end of the 8-week fixed-dose period for patients given clozapine (N=27) and those given haloperidol (N=25). The olanzapine group showed a significant increase of glucose levels at the end of the 6-week variable-dose period (N=22). Fourteen of the 101 patients developed abnormal glucose levels (>125 mg/dl) during the trial (six with clozapine, four with olanzapine, three with risperidone, and one with haloperidol). Cholesterol levels were increased at the end of the 8-week fixed-dose period for the patients given clozapine (N=27) and those given olanzapine (N=26); cholesterol levels were also increased at the end of the 6-week variable-dose period for patients given olanzapine (N=22). CONCLUSIONS: In this prospective randomized trial, clozapine, olanzapine, and haloperidol were associated with an increase of plasma glucose level, and clozapine and olanzapine were associated with an increase in cholesterol levels. The mean changes in glucose and cholesterol levels remained within clinically normal ranges, but approximately 14% of the patients developed abnormally high glucose levels during the course of their participation in the study. PMID- 12562577 TI - Subjective experience and D2 receptor occupancy in patients with recent-onset schizophrenia treated with low-dose olanzapine or haloperidol: a randomized, double-blind study. AB - OBJECTIVE: The authors tested the hypothesis that a dopamine D(2) receptor occupancy level between 60% and 70% in patients with recent-onset schizophrenia would result in optimal subjective experience. In addition, they sought preliminary evidence on whether subjective experience is better with low-dose olanzapine than with low-dose haloperidol. METHOD: Subjects (N=24) who met DSM-IV criteria for schizophrenia were randomly assigned to 6 weeks of double-blind treatment with either olanzapine, 7.5 mg/day, or haloperidol, 2.5 mg/day. Subjective experience, psychopathology, and extrapyramidal symptoms were assessed at baseline and at endpoint. After 6 weeks, D(2) receptor occupancy was assessed with [(123)I]iodobenzamide single photon emission computed tomography. RESULTS: The two study groups were similar at baseline. After 6 weeks, patients receiving olanzapine had a significantly lower mean dopamine D(2) receptor occupancy (51.0%, range=36%-67%) than those given haloperidol (65.5%, range=45%-75%). Receptor occupancy between 60% and 70% was associated with optimal subjective experience, and subjective experience improved significantly in the haloperidol group. CONCLUSIONS: A level of D(2) receptor occupancy between 60% and 70% is optimal for subjective experience of patients with recent-onset schizophrenia. Substantial interindividual variation in D(2) receptor occupancy was seen at fixed low-dose levels of olanzapine and haloperidol. Olanzapine, 7.5 mg/day, showed no superior subjective response over haloperidol, 2.5 mg/day. Olanzapine may need to be dosed higher than 7.5 mg/day for most patients with recent-onset schizophrenia, and haloperidol needs to be individually titrated in the very low dose range to reach optimal occupancy. PMID- 12562578 TI - Use of system-wide outcomes monitoring data to compare the effectiveness of atypical neuroleptic medications. AB - OBJECTIVE: Outcomes monitoring data in a large health care system were used to evaluate the effectiveness of atypical neuroleptic medications in actual clinical practice. METHOD: National administrative data from the Department of Veterans Affairs were used to identify patients with schizophrenia who had no change in antipsychotic medication over a 3-month index period in 1999 and for whom Global Assessment of Functioning Scale ratings were available from both the index period and the following year (N=9,066). Analysis of covariance, including potentially confounding factors, was used to compare change in functioning scores between patients who continued to take the same medication and those whose medication was switched during the follow-up year. Among patients whose medication was switched, change in functioning scores was compared by whether patients received a conventional neuroleptic or each of the four then available atypical neuroleptics after the medication switch. RESULTS: Patients who continued to take the same medication (N=7,157, 78.9%) had improved functioning scores from the index period to the follow-up year, while patients whose medication was switched (N=1,909, 21.1%) had declining functioning scores (mean change in scores of 0.6% and -3.7%, respectively). Among the patients whose medication was switched, no significant differences in functioning were found between patients grouped by the new type of medication received. CONCLUSIONS: In this large administrative database, patients with schizophrenia whose neuroleptic medication was switched showed significantly less improvement in Global Assessment of Functioning Scale scores than patients who continued to take their original medication. No significant differences in functioning scores were found for patients whose medication was switched to any of the atypical neuroleptics. PMID- 12562579 TI - Reward value of cigarette smoking for comparably heavy smoking schizophrenic, depressed, and nonpatient smokers. AB - OBJECTIVE: The study goal was to determine whether schizophrenic and depressed smokers perceive the reinforcement value of cigarette smoking differently from nonpsychiatric smokers who smoke as heavily. METHOD: The authors assessed the preferences for smoking cigarettes versus engaging in other pleasant activities, the perceived advantages and disadvantages of smoking, and the amount of reinforcement that would be needed to attain smoking abstinence among 26 schizophrenic, 26 depressed, and 26 nonpsychiatric heavy smokers. RESULTS: Both schizophrenic and depressed participants chose smoking as their preferred activity more often than nonpsychiatric smokers, and they did not differ from each other. The patients also exceeded the comparison group in the benefits they ascribed to smoking and felt they would require more incentives to quit, but they attributed comparable drawbacks to smoking. CONCLUSIONS: Schizophrenic and depressed smokers recognize many drawbacks associated with smoking, but compared to nonpatients who smoke as heavily, they also perceive more benefits and find cigarettes more appealing than alternative rewards. The heightened reward value of smoking warrants attention in tailoring tobacco control interventions for schizophrenic and depressed smokers. PMID- 12562580 TI - PET studies of the influences of nicotine on neural systems in cigarette smokers. AB - OBJECTIVE: The effects of acute nicotine administration and smoking on brain function were investigated in two studies, with the primary goal of identifying neural systems that mediate these effects. METHOD: In study 1, 18 healthy volunteer cigarette smokers were exposed to three conditions in a single session: 1) smoking a nicotine-containing cigarette, 2) smoking a denicotinized cigarette, or 3) receiving intravenous nicotine injections in conjunction with smoking a denicotinized cigarette. In study 2, 16 subjects smoked a nicotine-containing and denicotinized cigarette in each of two sessions 2 hours after receiving the nicotinic antagonist mecamylamine (10 mg) or placebo orally. Regional cerebral blood flow (rCBF) was assessed by using the bolus (15)O-labeled water method and positron emission tomography. Subjective measures of smoking withdrawal symptoms were also collected. RESULTS: A principal-components analysis of rCBF data pooled from the two studies identified three factors consisting of frontal, striatal, and reticular systems. The amygdala was considered as a separate region of interest. Nicotine increased normalized rCBF in the left frontal region and decreased rCBF in the left amygdala. The rCBF in the right hemisphere reticular system was related to nicotine dose in an inverted-U-shaped pattern and was strongly related to self-reported craving for cigarettes and to the addiction scale of a smoking motivation questionnaire. The effects of mecamylamine on rCBF were generally opposite to those of nicotine. CONCLUSIONS: The results indicate that nicotine influences brain regions involved in arousal and reward and suggest specific functional systems that may be linked to motivationally significant aspects of tobacco dependence. PMID- 12562581 TI - Inhibitory effect of hippocampal 5-HT1A receptors on human explicit memory. AB - OBJECTIVE: Recent studies have indicated that the serotonergic (5-HT) system plays important roles in memory function. However, the specific relationship between 5-HT(1A) receptors and memory function is not clear in the human brain. To clarify this relationship, the authors determined the availability of 5-HT(1A) receptors in the human brain and the relationship between regional receptor binding and memory function. METHOD: Using positron emission tomography (PET) with [(11)C]WAY-100635, the authors examined 5-HT(1A) receptors and assessed their relationship with memory function. The 5-HT(1A )agonist tandospirone was then administered to investigate the effect of 5-HT(1A) receptor stimulation on cognitive function and neuroendocrinological response. RESULTS: There was a significant negative correlation between explicit memory function and 5-HT(1A) receptor binding localized in the bilateral hippocampus where the postsynaptic 5 HT(1A) receptors are enriched. Furthermore, the administration of tandospirone dose-dependently impaired explicit verbal memory, while other cognitive functions showed no significant changes. The change in memory function paralleled those of body temperature and secretion of growth hormone, which were reported to be induced by the stimulation of postsynaptic 5-HT(1A) receptors. CONCLUSIONS: Postsynaptic 5-HT(1A )receptors localized in the hippocampal formation have a negative influence on explicit memory function, which raises the possibility that the antagonistic effect of postsynaptic 5-HT(1A) receptors in the hippocampus leads to improvement of human memory function. Drugs that work as antagonists on postsynaptic 5-HT(1A) receptors may be favorable for improved control of memory impairment. PMID- 12562582 TI - Delusional thoughts and regional frontal/temporal cortex metabolism in Alzheimer's disease. AB - OBJECTIVE: Delusional thoughts are common in patients with Alzheimer's disease and contribute prominently to morbidity. The pathophysiologic underpinnings for delusions in Alzheimer's disease are not well understood. In this study the authors examined the relationship between delusional thoughts and regional cortical metabolism in patients with Alzheimer's disease. METHOD: Twenty-five patients with probable Alzheimer's disease were included. None was taking psychotropic medication. Severity of delusions and other neuropsychiatric symptoms was assessed by using a semistructured interview and the Neurobehavioral Rating Scale just before the imaging procedure. [(18)F]Fluorodeoxyglucose positron emission tomography was used to measure resting cerebral glucose metabolic rates in the cortical lobes and in anatomically defined subregions of the frontal and temporal cortexes. RESULTS: A linear regression model, controlling for the effects of cognitive deficits, revealed a significant relationship between severity of delusional thought and the metabolic rates in three frontal regions: the right superior dorsolateral frontal cortex (Brodmann's area 8), the right inferior frontal pole (Brodmann's area 10), and the right lateral orbitofrontal region (Brodmann's area 47). Bivariate partial correlation analysis indicated that severity of delusions was associated with hypometabolism in additional prefrontal and anterior cingulate regions. Robust relationships with metabolism in regions of the temporal cortex were not apparent. CONCLUSIONS: Dysmetabolism in specific regions of the right prefrontal cortex may be associated with delusional thought in Alzheimer's disease. Delusions appear to reflect the pathophysiologic state of particular cortical regions. Activity across distributed neuronal networks and the specific content of delusional thoughts may modulate these relationships. PMID- 12562583 TI - Fatigue in obstructive sleep apnea: driven by depressive symptoms instead of apnea severity? AB - OBJECTIVE: Obstructive sleep apnea is a common and frequently devastating illness that often includes significant fatigue. Fatigue is also a hallmark depressive symptom. The authors wondered if depressive symptoms in patients with obstructive sleep apnea would account for some of the fatigue beyond that explained by obstructive sleep apnea severity. METHOD: Sixty patients with obstructive sleep apnea-i.e., score >/=15 on the respiratory disturbance index (mean score=49; range=15-111)-underwent polysomnography and completed the Center for Epidemiological Studies Depression Scale (CES-D Scale), Profile of Mood States (POMS), and Medical Outcomes Study surveys. Data were analyzed by using hierarchical regression, with POMS fatigue score as the dependent variable (step 1, forced entry of apnea severity variables; step 2, forced entry of CES-D Scale score). RESULTS: Whereas score on the respiratory disturbance index and the percent of time oxygen saturation was <90% together accounted for 4.2% of variance in scores on the POMS fatigue scale, the CES-D Scale score accounted for 10 times the variance (i.e., an additional 42.3%) in POMS fatigue scale score. CONCLUSIONS: After obstructive sleep apnea severity was controlled, higher levels of depressive symptoms were dramatically and independently associated with greater levels of fatigue. Assessment and treatment of mood symptoms-not just treatment of the disordered breathing itself-might reduce the fatigue experienced by patients with obstructive sleep apnea. PMID- 12562584 TI - The Declaration of Helsinki and clinical trials: a focus on placebo-controlled trials in schizophrenia. AB - OBJECTIVE: The authors' goal was to consider ethical approaches to placebo controlled clinical trials in the light of the evolving Declaration of Helsinki, with special attention to applications to research on schizophrenia. METHOD: They review the Helsinki position on placebos, including the 2002 Clarification, exploring the potential negative effects of banning placebos in studies involving conditions for which at least partially effective treatments exist. The Clarification is examined as an approach to this issue that, in contrast to earlier formulations, better acknowledges the complexity of clinical research and the need for protocol-specific determinations. Placebo controls in schizophrenia studies are used to illustrate issues relevant to all clinical research on therapeutic interventions. RESULTS: The Helsinki Clarification provides a basis for operationalizing criteria for review of placebo use in clinical trials. Six criteria are proposed for judging the ethical acceptability of placebo controls, including the likelihood that the intervention being tested will have clinically significant advantages over existing treatments, the presence of compelling reasons for placebo use, subject selection that minimizes the possibility of serious adverse consequences, and a risk-versus-benefit analysis that favors the advantages from placebo use over the risks to subjects. CONCLUSIONS: The Helsinki Clarification constitutes an important advance in international approaches to placebo use, requiring protocol-by-protocol judgments on complex issues of clinical research ethics. When operationalized, it provides review boards with a useful methodology for reaching determinations on the appropriateness of placebo controls in particular studies. PMID- 12562585 TI - Are top journals biased against eating disorders topics? AB - OBJECTIVE: This study investigated whether there is a bias against eating disorders research among the leading psychiatric, psychological, and medical journals. METHOD: The authors performed a comparison between the number of empirical articles published about anorexia nervosa and/or bulimia nervosa and the number of articles published about panic disorder and/or agoraphobia (i.e., disorders of comparable disease burden) in 29 high-impact journals over a 5-year period (1996-2001). RESULTS: There were almost twice as many published empirical articles about panic disorder and/or agoraphobia (N=365) as there were about anorexia nervosa and/or bulimia nervosa (N=169). CONCLUSIONS: The findings indicate a possible bias against eating disorders research among some leading psychiatric journals. Alternative explanations and implications are discussed. PMID- 12562586 TI - The relation between eating disorders and components of perfectionism. AB - OBJECTIVE: The authors' goal was to explore the relation between perfectionism and psychopathology, including eating disorders. METHOD: Using logistic regression, the authors calculated odds ratios for the associations between perfectionism subscale scores and psychiatric disorders in 1,010 female twins who completed the Multidimensional Perfectionism Scale and participated in diagnostic interviews. RESULTS: Elevated concern over mistakes was associated with anorexia and bulimia nervosa but not with other psychiatric disorders. Doubts about actions was associated with eating and anxiety disorders. Multivariable models confirmed that higher scores on the subscales for concern over mistakes and doubts about actions were most strongly associated with eating disorders. CONCLUSIONS: The aspect of perfectionism captured by scores on a subscale measuring concern over mistakes may be particularly associated with eating disorders and not generically predictive of psychopathology. PMID- 12562587 TI - Implications of childhood sexual abuse for adult borderline personality disorder and complex posttraumatic stress disorder. AB - OBJECTIVE: This study examined whether women with a history of early-onset sexual abuse or those with late-onset sexual abuse were more likely to meet diagnostic criteria for both borderline personality disorder and complex posttraumatic stress disorder (PTSD). METHOD: The Revised Diagnostic Interview for Borderlines and the Trauma Assessment Package were administered to 65 women from three outpatient clinics in a metropolitan area. Thirty-eight subjects met criteria for early-onset abuse, while 27 subjects met criteria for late-onset abuse. RESULTS: The diagnoses of both borderline personality disorder and complex PTSD were significantly higher in women reporting early-onset abuse than in those with late onset abuse. The trauma variables sexual abuse and paternal incest were significant predictors of both diagnoses. CONCLUSIONS: In contrast to those with comorbid diagnoses, some women with a history of childhood sexual abuse may be extricated from the diagnosis of borderline personality disorder and subsumed under that of complex PTSD. PMID- 12562588 TI - Reduction of nightmares and other PTSD symptoms in combat veterans by prazosin: a placebo-controlled study. AB - OBJECTIVE: Prazosin is a centrally active alpha(1) adrenergic antagonist. The authors' goal was to evaluate prazosin efficacy for nightmares, sleep disturbance, and overall posttraumatic stress disorder (PTSD) in combat veterans. METHOD: Ten Vietnam combat veterans with chronic PTSD and severe trauma-related nightmares each received prazosin and placebo in a 20-week double-blind crossover protocol. RESULTS: Prazosin (mean dose=9.5 mg/day at bedtime, SD=0.5) was superior to placebo for the three primary outcome measures: scores on the 1) recurrent distressing dreams item and the 2) difficulty falling/staying asleep item of the Clinician-Administered PTSD Scale and 3) change in overall PTSD severity and functional status according to the Clinical Global Impression of change. Total score and symptom cluster scores for reexperiencing, avoidance/numbing, and hyperarousal on the Clinician-Administered PTSD Scale also were significantly more improved in the prazosin condition, and prazosin was well tolerated. CONCLUSIONS: These data support the efficacy of prazosin for nightmares, sleep disturbance, and other PTSD symptoms. PMID- 12562589 TI - Changes in regional cerebral blood flow with venlafaxine in the treatment of major depression. AB - OBJECTIVE: Neuroimaging studies reveal abnormalities of regional cerebral blood flow (rCBF) in major depression. In this study the authors prospectively investigated rCBF and clinical response to venlafaxine, a novel antidepressant. METHOD: A trial of venlafaxine was performed with seven patients referred with ICD-10 major depression. At entry and 6-week follow-up, the Beck Depression Inventory and Hamilton Depression Rating Scale were administered and rCBF was measured by means of single photon emission computed tomography with [(99m)Tc]hexamethylpropyleneamine oxime. Blood flow changes were explored with statistical parametric mapping. RESULTS: The subjects showed significant improvement after treatment. Statistical parametric mapping analysis revealed increased rCBF bilaterally in the thalamus and decreased rCBF in the left occipital lobe, right cerebellum, and temporal cortex bilaterally. CONCLUSIONS: These data confirm limbic cortical rCBF changes associated with effective antidepressant treatment. PMID- 12562590 TI - Differentiation of geriatric major depression from Alzheimer's disease with CSF tau protein phosphorylated at threonine 231. AB - OBJECTIVE: Differentiation of geriatric major depression from Alzheimer's disease is hampered by overlapping symptoms. Increased CSF concentrations of tau protein phosphorylated at threonine 231 (p-tau(231)) have been suggested as a biomarker for Alzheimer's disease. The authors asked whether p-tau(231) levels improve the differential diagnosis between geriatric major depression and Alzheimer's disease. METHOD: Included were 34 depression subjects, 64 with probable Alzheimer's disease, 17 with possible Alzheimer's disease, and 21 healthy comparison subjects. P-tau(231) concentrations were measured with an enzyme linked immunosorbent assay. RESULTS: P-tau(231) levels were significantly higher in Alzheimer's disease than in geriatric major depression patients and healthy comparison subjects. For differentiation of probable Alzheimer's disease from major depression, p-tau(231) correctly allocated 87% of subjects. When possible mild Alzheimer's disease was compared to major depression, p-tau(231) correctly allocated 78% of subjects. CONCLUSIONS: CSF p-tau(231) should be evaluated as a potential biological marker for differentiation of geriatric depression from Alzheimer's disease. PMID- 12562591 TI - Parahippocampal volume deficits in subjects with aging-associated cognitive decline. AB - OBJECTIVE: Neuropathological evidence suggests that the earliest changes in Alzheimer's disease selectively affect the parahippocampal regions of the brain. This study was conducted to determine if otherwise healthy elderly subjects with mild cognitive impairment had structural volume deficits affecting the parahippocampal gyrus. METHOD: Magnetic resonance imaging (MRI) was used to compare global and regional brain volumes in 21 subjects with mild cognitive deficits defined according to the criteria for aging-associated cognitive decline, 22 cognitively intact comparison subjects, and 12 patients with Alzheimer's disease. RESULTS: Compared with the cognitively intact subjects, the subjects with aging-associated cognitive decline had a significantly smaller mean volume of the right parahippocampal gyrus. The subjects with aging-associated cognitive decline had a mean parahippocampal volume that was intermediate between that of the Alzheimer's disease patients and that of the cognitively intact subjects. CONCLUSIONS: Parahippocampal atrophy underlies the observed cognitive deficits in aging-associated cognitive decline. These findings support the hypothesis that aging-associated cognitive decline represents a preclinical stage of Alzheimer's disease. PMID- 12562592 TI - Older patients with schizophrenia: nature of dwelling status and symptom severity. AB - OBJECTIVE: This cross-sectional study enrolled elderly patients with diagnoses of schizophrenia or schizoaffective disorder. METHOD: The 85 subjects were dichotomized into two groups on the basis of dwelling status: those living independently (N=35) and those living in residential settings (N=50). The groups were compared with regard to scores on the Mini-Mental State Examination (MMSE), the Brief Psychiatric Rating Scale, the Scale for the Assessment of Negative Symptoms (SANS), the Geriatric Depression Scale and by age. RESULTS: Patients living independently had significantly higher MMSE scores, lower SANS scores, more years of education, and were younger than the patients living in residential settings. CONCLUSIONS: These data suggest that although cognition, negative symptoms, and age are important discriminators with regard to dwelling status, cognition and negative symptoms appear to have the strongest impact. PMID- 12562594 TI - Electrolyte-balanced sports drink for polydipsia-hyponatremia in schizophrenia. PMID- 12562593 TI - Omega-3 fatty acids for depression in pregnancy. PMID- 12562595 TI - Biperiden for excessive sweating from methadone. PMID- 12562597 TI - Change in brain function with placebo. PMID- 12562598 TI - Real-life research. PMID- 12562599 TI - Diabetes and atypical neuroleptics. PMID- 12562600 TI - Hagiographic treatment of C.G. Jung. PMID- 12562601 TI - Diabetes and atypical neuroleptics. PMID- 12562602 TI - Brain changes and placebo. PMID- 12562606 TI - Misdiagnosis of conversion disorder. PMID- 12562608 TI - Symptom factors and clinical subtypes in mania. PMID- 12562609 TI - Physical anomalies and schizophrenia spectrum disorders. PMID- 12562610 TI - Affective instability in personality disorders. PMID- 12562625 TI - Carcinogenesis bioassays and protecting public health. Commemorating the lifework of Cesare Maltoni and colleagues. Conference proceedings. April 29-30, 2002, New York, New York, USA. PMID- 12562626 TI - Remembering Professor Cesare Maltoni. AB - Professor Cesare Maltoni, a renowned leader in the research of the hazards of carcinogens in the workplace, died on January 22, 2001 at the age of 70. Born in Faenza (Ravenna), Italy on November 17, 1930, he received his M.D. degree from the University of Bologna in 1954-1955. He was Director of the Institute of Oncology of Bologna (1964 to 1997), Director of the Bologna Centre for the Prevention and Detection of Tumours and Oncological Research (1966 to 1989), and Scientific Director, European Foundation of Oncology and Environmental Sciences "B. Ramazzini" from 1993 until he died. Maltoni conducted long-term carcinogenic studies on some 200 agents. He was the first to demonstrate that vinyl chloride is a carcinogen that produces, among other tumors, angiosarcoma of the liver. He was the first to show that benzene is a powerful multipotential carcinogen. Maltoni authored more than 700 original scientific publications, books, and proceedings. He was editor and coeditor of many journals. Among his many awards were the Stokinger Award, American Conference of Governmental Industrial Hygienists (ACGIH), Kansas City, 1995; International Award "B. Ramazzini" of the Collegium Ramazzini, Washington, 1995; International I.J. Selikoff Memorial Award, Washington, 1995; and the Sigillum Magnum of the University of Bologna, 1997. Many fellow Ramazzinians and his coworkers wish to be considered part of the group of, as he once wrote, his "always family friends" and to remember happy moments when we were together. A man of great stature and many contributions, Cesare Maltoni will never be forgotten. PMID- 12562627 TI - Ramazzini Foundation cancer program: history and major projects, life-span carcinogenicity bioassay design, chemicals studied, and results. AB - The Ramazzini Foundation research program was started over thirty years ago. The features of this program are: (1) systematic and integrated project design; (2) consistency over time; (3) homogeneity of approach: key members of the team remain unchanged; and (4) choice to work on new frontiers of scientific research. The program centers mainly on three projects: Project 1: experimental carcinogenicity bioassays; Project 2: experimental anticarcinogenesis assays to identify factors and active principles (compounds) capable of opposing the onset of tumors while being suitable for preventive/chemopreventive intervention; Project 3: epidemiological studies, both descriptive and analytical, on tumor incidence and mortality in persons professionally and environmentally exposed to industrial carcinogenic risks. The project involving experimental carcinogenicity bioassays for the identification of exogenous carcinogens (environmental and industrial above all) began in 1966. This project has included 398 experimental bioassays on 200 compounds/agents using some 148,000 animals monitored until their spontaneous death. Among the studies already concluded, 47 agents have shown "clear evidence" of carcinogenicity. The results have demonstrated for the first time that (1) vinyl chloride can cause liver angiosarcoma as well as other tumors; (2) benzene is carcinogenic in experimental animals for various tissues and organs; (3) formaldehyde may produce lymphomas and leukemias; and (4) methyl tert-butyl ether (MTBE), the most common oxygenated additive used in gasolines, can cause lymphomas/leukemias. Many of the results achieved have led to the introduction of norms and measures of primary prevention. PMID- 12562628 TI - Results of long-term experimental studies on the carcinogenicity of methyl alcohol and ethyl alcohol in rats. AB - Methyl alcohol was administered in drinking water supplied ad libitum at doses of 20,000, 5,000, 500, or 0 ppm to groups of male and female Sprague-Dawley rats 8 weeks old at the start of the experiment. Animals were kept under observation until spontaneous death. Ethyl alcohol was administered by ingestion in drinking water at a concentration of 10% or 0% supplied ad libitum to groups of male and female Sprague-Dawley rats; breeders and offspring were included in the experiment. Treatment started at 39 weeks of age (breeders), 7 days before mating, or from embryo life (offspring) and lasted until their spontaneous death. Under tested experimental conditions, methyl alcohol and ethyl alcohol were demonstrated to be carcinogenic for various organs and tissues. They must also be considered multipotential carcinogenic agents. In addition to causing other tumors, ethyl alcohol induced malignant tumors of the oral cavity, tongue, and lips. These sites have been shown to be target organs in man by epidemiologic studies. PMID- 12562629 TI - Results of long-term carcinogenicity bioassays on tert-amyl-methyl-ether (TAME) and di-isopropyl-ether (DIPE) in rats. AB - Tert-amyl-methyl ether (TAME) was administered by gavage in extra virgin olive oil solution at concentrations of 750, 250, or 0 mg/kg bw to groups of 100 male and 100 female Sprague-Dawley rats 8 weeks old at the start of the experiment. Di isopropyl ether (DIPE) was administered in the same manner at the doses of 1000, 250, or 0 mg/kg body weight to groups of 100 male and 100 female Sprague-Dawley rats. TAME and DIPE were each delivered in 1-mL solution 4 days a week for 78 weeks. Control animals received 1 mL of extra virgin olive oil without TAME or DIPE. At the end of the treatment period, all animals were kept under observation until spontaneous death. Under these test conditions, TAME and DIPE were found to be potential carcinogenic agents for various organs and tissues. PMID- 12562630 TI - Results of long-term experimental studies on the carcinogenicity of formaldehyde and acetaldehyde in rats. AB - Formaldehyde was administered for 104 weeks in drinking water supplied ad libitum at concentrations of 1500, 1000, 500, 100, 50, 10, or 0 mg/L to groups of 50 male and 50 female Sprague-Dawley rats beginning at seven weeks of age. Control animals (100 males and 100 females) received tap water only. Acetaldehyde was administered to 50 male and 50 female Sprague-Dawley rats beginning at six weeks of age at concentrations of 2,500, 1,500, 500, 250, 50, or 0 mg/L. Animals were kept under observation until spontaneous death. Formaldehyde and acetaldehyde were found to produce an increase in total malignant tumors in the treated groups and showed specific carcinogenic effects on various organs and tissues. PMID- 12562631 TI - Results of long-term carcinogenicity bioassay on vinyl acetate monomer in Sprague Dawley rats. AB - Vinyl acetate monomer (VAM) was administered in drinking water supplied ad libitum at doses of 5,000, 1,000, and 0 ppm (v/v) to 17-week-old Sprague-Dawley rats (breeders) and to 12-day embryos (offspring). Treatment lasted for 104 weeks; thereafter, animals were kept under control conditions until spontaneous death. VAM was found to cause an increase in total malignant tumors and in carcinomas and/or precursor lesions of the oral cavity, lips, tongue, esophagus, and forestomach. Based on these data, VAM must be considered a multipotent carcinogen. PMID- 12562632 TI - Results of long-term experimental studies on the carcinogenicity of ethylene-bis dithiocarbamate (Mancozeb) in rats. AB - Mancozeb, an ethylene-bis-dithiocarbamate (EBDC), has been one of the most commonly used fungicides in commercial use for several decades. Nevertheless, up to now, no adequate published experimental studies on the carcinogenicity of Mancozeb have been published. Because of the importance of the compound and of the number of people potentially exposed (workers engaged in the production and use of the fungicide, people living in agricultural areas where the compound is sprayed, and people consuming polluted products), a long-term experimental study of Mancozeb was begun at the Cancer Research Center of the Ramazzini Foundation. Groups of 150 male and female Sprague-Dawley rats, 8 weeks old at the start of the treatment, were administered Mancozeb at the concentration of 1000, 500, 100, 10, and 0 ppm in feed supplied ad libitum for 104 weeks. At the end of the treatment, animals were kept under controlled conditions until spontaneous death. Mancozeb caused an increase in (1) total malignant tumors, (2) malignant mammary tumors, (3) Zymbal gland and ear duct carcinomas, (4) hepatocarcinomas, (5) malignant tumors of the pancreas, (6) malignant tumors of the thyroid gland, (7) osteosarcomas of the bones of the head, and (8) hemolymphoreticular neoplasias. On the basis of these data, Mancozeb must be considered a multipotent carcinogenic agent. PMID- 12562633 TI - Carcinogenic effects of benzene: Cesare Maltoni's contributions. AB - Cesare Maltoni's contributions to understanding, identifying, and characterizing widely used commercial chemicals in experimental animals are among the most important methods developed in the history of toxicology and serve to protect working men and women, the general population, and our environment from hazardous substances. Maltoni developed experimental methods that have reached the "platinum standard" for protection of public health. Benzene was among the 400 or more chemicals that Maltoni and his associates tested for carcinogenicity. In 1976, Maltoni reported that benzene is a potent experimental carcinogen. Maltoni's experiments clearly demonstrated that benzene is carcinogenic in Sprague-Dawley rats, Wistar rats, Swiss mice, and RF/J mice when administered by inhalation or ingestion. Benzene caused carcinomas of the Zymbal gland, oral cavity, nasal cavities; cancers of the skin, forestomach, mammary glands, and lungs; angiosarcomas and hepatomas of the liver; and hemolymphoreticular cancers. Thus, benzene was shown to be a multipotential carcinogen that produced cancers in several species of animals by various routes of administration. On November 2, 1977, Chemical Week reported that Maltoni provided a "bombshell" when he demonstrated the "first direct link" between benzene and cancer. In this paper, I shall summarize early experiments and human studies and reports; Maltoni's experimental contribution to understanding the carcinogenicity of benzene in humans and animals; earlier knowledge concerning benzene toxicity; and benzene standards and permissible exposure levels. PMID- 12562634 TI - Carcinogenicity of methyl-tertiary butyl ether in gasoline. AB - Methyl tertiary butyl ether (MTBE) was added to gasoline on a nationwide scale in 1992 without prior testing of adverse, toxic, or carcinogenic effects. Since that time, numerous reports have appeared describing adverse health effects of individuals exposed to MTBE, both from inhalation of fumes in the workplace and while pumping gasoline. Leakage of MTBE, a highly water-soluble compound, from underground storage tanks has led to contamination of the water supply in many areas of the United States. Legislation has been passed by many states to prohibit the addition of MTBE to gasoline. The addition of MTBE to gasoline has not accomplished its stated goal of decreasing air pollution, and it has posed serious health risks to a large portion of the population, particularly the elderly and those with respiratory problems, asthma, and skin sensitivity. Reports of animal studies of carcinogenicity of MTBE began to appear in the 1990s, prior to the widespread introduction of MTBE into gasoline. These reports were largely ignored. In ensuing years, further studies have shown that MTBE causes various types of malignant tumors in mice and rats. The National Toxicology Program (NTP) Board of Scientific Counselors' Report on Carcinogens Subcommittee met in December 1998 to consider listing MTBE as "reasonably anticipated to be a human carcinogen." In spite of recommendations from Dr. Bailer, the primary reviewer, and other scientists on the committee, the motion to list MTBE in the report was defeated by a six to five vote, with one abstention. On the basis of animal studies, it is widely accepted that if a chemical is carcinogenic in appropriate laboratory animal test systems, it must be treated as though it were carcinogenic in humans. In the face of compelling evidence, NTP Committee members who voted not to list MTBE as "reasonably anticipated to be a human carcinogen" did a disservice to the general public; this action may cause needless exposure of many to health risks and possibly cancers. PMID- 12562635 TI - Asbestos fibers contributing to the induction of human malignant mesothelioma. AB - To elucidate the features of the asbestos fibers contributing to the induction of human malignant mesothelioma, we used high-resolution analytical electron microscopy to determine the type, number, and dimensions of asbestos fibers in lung and mesothelial tissues in 168 cases of mesothelioma. RESULTS: 1. Asbestos fibers were present in almost all of the lung and mesothelial tissues from the mesothelioma cases. 2. The most common types of asbestos fibers in lung were either an admixture of chrysotile with amphiboles, amphibole alone, and occasionally chrysotile alone. In mesothelial tissues, most asbestos fibers were chrysotile. 3. In lung, amosite fibers were greatest in number followed by chrysotile, crocidolite, tremolite/actinolite, and anthophyllite. In mesothelial tissues, chrysotile fibers were 30.3 times more common than amphiboles. 4. In some mesothelioma cases, the only asbestos fibers detected in either lung or mesothelial tissue were chrysotile fibers. 5. The average number of asbestos fibers in both lung and mesothelial tissues was two orders of magnitude greater than the number found in the general population. 6. The majority of asbestos fibers in lung and mesothelial tissues were shorter than 5 micro m in length. CONCLUSIONS: 1) Fiber analysis of both lung and mesothelial tissues must be done to determine the types of asbestos fibers associated with the induction of human malignant mesothelioma; 2) short, thin asbestos fibers should be included in the list of fiber types contributing to the induction of human malignant mesothelioma; 3) RESULTS support the induction of human malignant mesothelioma by chrysotile. PMID- 12562636 TI - Carcinogenicity and mechanistic insights on the behavior of epoxides and epoxide forming chemicals. AB - Many epoxides and their precursors are high production volume chemicals that have major uses in the polymer industry and as intermediates in the manufacture of other chemicals. Several of these chemicals were demonstrated to be carcinogenic in laboratory animal studies conducted by the Ramazzini Foundation (e.g., vinyl chloride, acrylonitrile, styrene, styrene oxide, and benzene) and by the National Toxicology Program (e.g., ethylene oxide, 1,3-butadiene, isoprene, chloroprene, acrylonitrile, glycidol, and benzene). The most common sites of tumor induction were lung, liver, harderian gland, and circulatory system in mice; Zymbal's gland and brain in rats; and mammary gland and forestomach in both species. Differences in cancer outcome among studies of epoxide chemicals may be related to differences in study design (e.g., dose, duration, and route of exposure; observation period; animal strains), as well as biological factors affecting target organ dosimetry of the DNA-reactive epoxide (toxicokinetics) and tissue response (toxicodynamics). N7-Alkylguanine, N1-alkyladenine, and cyclic etheno adducts, as well as K-ras and p53 mutations, have been detected in animals and/or workers exposed to several of these chemicals. The classifications of these chemical carcinogens by IARC and NTP are based on animal and human data and results of mechanistic studies. Reducing occupational and environmental exposures to these chemicals will certainly reduce human cancer risks. PMID- 12562637 TI - Primary prevention protects public health. AB - It is widely accepted that epidemiological data provide the only reliable evidence of a carcinogenic effect in humans, but epidemiology is unable to provide early warning of a cancer risk. The experimental approach to carcinogenicity can ascertain and predict potential cancer risks to humans in time for primary prevention to be successful. Unfortunately, only in rare instances were experimental data considered sufficiently convincing per se to stimulate the adoption of preventive measures. The experimental testing of environmental agents is the second line of defense against potential human carcinogens. The first line is the testing of synthesized agents, be these pesticides, medical drugs, or industrial chemical/physical agents, at the time of their development. We do not know, however, how many substances have been prevented from entering the environment because most tests are carried out by commercial or private laboratories and results are rarely released. A better understanding of the mechanisms underlying the sequence of events of the carcinogenesis process will eventually lead to a more accurate characterization and quantification of risks. However, the ways that mechanistic data have been used lately for evaluating evidence of carcinogenicity have not necessarily meant that the evaluations were more closely oriented toward public health. A tendency has surfaced to dismiss the relevance of long-term carcinogenicity studies. In the absence of absolute certainty, rarely if ever reached in biology, it is essential to adopt an attitude of responsible caution, in line with the principles of primary prevention, the only one that may prevent unlimited experimentation on the entire human species. PMID- 12562638 TI - The National Toxicology Program rodent bioassay: designs, interpretations, and scientific contributions. AB - The National Toxicology Program rodent cancer bioassay program design evolved from that of the National Cancer Institute in the 1970s. Groups of 50 or more mice are assigned to control or treatment groups. Test substances are given at three dose levels by intubation, dietary or drinking water consumption, or dermal or inhalation exposure. Dosing starts at age 5-6 weeks and lasts for 2 years, when surviving animals receive a complete histopathologic examination. Statistical approaches accommodate survival differences and no longer require differentiation between fatal and incidental tumors. Photocarcinogenicity studies, employing SKH-1 hairless mice, evaluate onset of skin papillomas and incidences at 1 year. Top doses are chosen to expose animals to a minimally toxic challenge and lower doses to operate within the linear range of kinetics. This dosing allows comparison of results across studies. Bioassay and ancillary studies successfully identify tumor-causing agents in rodents, provide information on dose-response, and characterize other chemical-related toxicities. NTP and Ramazzini Foundation bioassay designs differ in several aspects, but bioassays at both institutions provide chemical-specific information for predicting human carcinogens, thus providing for protection of public health. Bioassays constitute an essential information reference set for new assay development and further investigations into mechanisms of action. The scientific community and the public owe a huge debt of gratitude to Dr. Cesare Maltoni of the European Foundation of Oncology and Environmental Sciences and to Dr. David P. Rall of the National Institute of Environmental Health Sciences for their foresight and wisdom in creating and nurturing these bioassay programs. PMID- 12562639 TI - Chemicals studied and evaluated in long-term carcinogenesis bioassays by both the Ramazzini Foundation and the National Toxicology Program: in tribute to Cesare Maltoni and David Rall. AB - The Ramazzini Foundation (RF) in Bentivoglio, Italy and the National Toxicology Program (NTP) in Research Triangle Park, North Carolina have carried out several hundred chemical carcinogenesis bioassays: 200 by RF and 500 by NTP. Of these, 21 have been evaluated by both laboratories. The 14 chemicals for which both laboratories have designed, conducted, and reported bioassay results are: acrylonitrile, benzene, chlorine, diesel fuel, ethylbenzene, methylene chloride (dichloromethane), propylene, styrene, styrene oxide, toluene, trichloroethylene, trichlorofluoromethane, vinylidene chloride, and xylenes. The other seven chemicals (two are fibers) were evaluated by both laboratories, but results have not yet been published. Results of these 14 interlaboratory studies were compared both to explore consistency of carcinogenic responses and to identify possible factors that may reveal reasons for any differences observed. Individual carcinogenesis results from each laboratory were duplicated and complementary. Of the 14 chemicals compared, 11 (80%) were either carcinogenic (9 chemicals) or noncarcinogenic (2 chemicals) in both studies. Eight of the paired chemicals had at least one carcinogenic target site in common. The other three were carcinogenic in one laboratory but not in the other. Possible explanations for these differences include dose, method of administration, duration of follow-up, and whether or not total tumors are counted. The collaboration between these two pioneering bioassay laboratory programs contributes greatly to our understanding of chemical carcinogenesis and results in better protection of workers and the general population from chemical diseases, especially cancers. PMID- 12562640 TI - Chemotherapy in gastric cancer: a never ending saga. PMID- 12562641 TI - Prediction of response to therapy by biomolecular markers: from the research laboratory to the clinic. PMID- 12562642 TI - The pharmacological treatment of aggressive fibromatosis: a systematic review. AB - BACKGROUND: Despite the use of surgery and radiotherapy, 20-35% of patients with aggressive fibromatosis (AF) will have local recurrence. The purpose of this review was to collect and analyze all available information regarding the role of non-cytotoxic and cytotoxic chemotherapy in AF that has been accumulated over the past few decades. PATIENTS AND METHODS: A systematic review of published clinical trials, studies and case series was carried out using the Medline Express Databases and the Cochrane Collaboration Database from 1970 to October 2000. RESULTS: Most studies published in the literature are in the form of successful case reports and single-arm series with small patient numbers. Most commonly used agents include hormonal agents, non-steroidal anti-inflammatory drugs (NSAIDs), interferons and cytotoxics. The literature data support the use of hormonal agents. Several questions, however, remain unresolved, such as which is the most suitable endocrine manipulation and what is the optimal dose and duration of treatment. NSAIDs and interferons have demonstrated activity against AF either alone or in combination with hormone therapy or chemotherapy but the precise mechanism of action is still unknown. Finally, there is growing evidence in the literature that chemotherapy is effective against AF with almost one in two patients being likely to respond. CONCLUSIONS: The evidence in the literature supports the opinion that both non-cytotoxic and cytotoxic chemotherapies are effective against AF. However, the lack of sufficient patient numbers and randomized trials compromises the validity of the reported results and mandates further investigation with properly designed prospective studies including larger patient numbers, with main end points to include not only tumor response rate and survival but also quality-of-life issues. PMID- 12562643 TI - The unknown biology of the unknown primary tumour: a literature review. AB - The unknown primary tumour (UPT) is an intriguing clinical phenomenon found in approximately 5% of all newly diagnosed patients with cancer. It is unclear whether UPT forms a distinct biological entity with specific genetic and phenotypic characteristics, or whether it is merely a clinical presentation of metastases in patients in whom the primary tumour cannot be detected and does not result in any visible clinical signs. Understanding the basic biology of UPT may shed light on this issue and, moreover, may have a direct impact on clinical care. A review of the literature revealed only a limited number of publications describing the genetic and phenotypic features of UPT, most of which focus only on the potential of these markers to predict prognosis. The question as to whether the biology of UPT is different from tumours of known primaries therefore remains unanswered. Further insight into the molecular mechanisms underlying the oncogenesis of UPT, e.g. by applying newly available DNA and gene profiling microarray techniques, will be necessary to understand its specific biology and to develop more effective treatments. PMID- 12562644 TI - Weekly oral paclitaxel as first-line treatment in patients with advanced gastric cancer. AB - BACKGROUND: Pharmacokinetic study has shown that co-administration of cyclosporin A (CsA), which acts as a P-glycoprotein (P-gp) and CYP-3A blocker, resulted in an 8-fold increase in the systemic exposure of oral paclitaxel. Two doses of oral paclitaxel on 1 day in combination with CsA resulted in higher systemic exposure than single dose administration. PATIENTS AND METHODS: In this phase II study, chemonaive patients with advanced gastric cancer received oral paclitaxel weekly in two doses of 90 mg/m(2) on the same day; CsA (10 mg/kg) was given 30 min before each dose of oral paclitaxel. RESULTS: In 25 patients, the main toxicities were: nausea CTC grade 2/3, 10 patients (40%); vomiting grade 2/3, 4 patients (20%); diarrhea grade 2/3, 6 patients (24%); neutropenia grade 3/4, 5 patients (20%). In the 24 evaluable patients, eight partial responses were observed, resulting in an overall response rate (ORR) of 33% [95% confidence interval (CI) 18% to 52%]. Eleven patients had stable disease (46%) and 5 patients showed progressive disease (21%). The ORR in the total population was 32% (95% CI 17% to 50%). The median time to progression was 16 weeks (95% CI 9-22). Pharmacokinetic analyses revealed that the mean area under the plasma concentration-time curve (AUC) of orally administered paclitaxel (+/- standard deviation) was 3757.6 +/- 939.4 ng.h/ml in week 1 and 3928.4 +/- 1281 ng.h/ml in week 2. The intrapatient variability in the AUC was 12%. CONCLUSIONS: Oral paclitaxel in combination with CsA is both active and safe in chemonaive patients with advanced gastric cancer. Toxicities were mainly gastrointestinal. PMID- 12562645 TI - Weekly gemcitabine and cisplatin chemotherapy: a well-tolerated but ineffective chemotherapeutic regimen in advanced pancreatic cancer patients. A report from the Italian Group for the Study of Digestive Tract Cancer (GISCAD). AB - BACKGROUND: This phase II study was initiated to determine the activity and toxicity of a combination of gemcitabine (GEM) and cisplatin (CDDP) in patients with pancreatic cancer. PATIENTS AND METHODS: CDDP 35 mg/m(2) was given as a 30 min infusion and GEM 1000 mg/m(2) as a 30-min infusion. Both drugs were administered once weekly for 2 consecutive weeks out of every 3 weeks to chemonaive patients with locally advanced or metastatic pancreatic cancer. RESULTS: Forty-five advanced pancreatic cancer patients received this regimen for a total of 180 cycles of chemotherapy. One complete and four partial responses have been observed for an overall response rate of 9% (95% confidence interval 10% to 11%). Twenty-one patients (46%) had stable disease and 19 progressed on therapy. The median time to progression was 3.6 months, with a median survival of 5.6 months. A clinical benefit was obtained in nine of 37 patients (24%). Side effects were mainly represented by hematological toxicity. Grade 3/4 WHO toxicities included neutropenia (6% of the patients) and thrombocytopenia (11%). The dose of GEM and CDDP was reduced in 14 patients (31%) and treatment was delayed in 10 patients (22%). CONCLUSIONS: Our results in terms of response rate, clinical benefit and survival do not support an advantage for the combination of GEM and CDDP given by this schedule. PMID- 12562646 TI - Cigarette tar yield and risk of upper digestive tract cancers: case-control studies from Italy and Switzerland. AB - BACKGROUND: Tobacco smoking is one of the main risk factors for oral, pharyngeal and oesophageal cancers in developed countries. Information on the role of the tar yield of cigarettes in upper digestive tract carcinogenesis is sparse and needs to be updated because the tar yield of cigarettes has steadily decreased over the last few decades. PATIENTS AND METHODS: We analysed two case-control studies, from Italy and Switzerland, conducted between 1992 and 1999, involving 749 cases of oral and pharyngeal cancer and 1770 controls, and 395 cases of squamous-cell oesophageal carcinoma and 1066 matched controls. Odds ratios (ORs) were estimated by unconditional multiple logistic regression models, including terms for age, sex, study centre, education and alcohol consumption. RESULTS: Based on the brand of cigarettes smoked for the longest time, the multivariate ORs for current smokers compared with never smokers were 6.1 for <20 mg and 9.8 for >or=20 mg tar for oral and pharyngeal neoplasms, and 4.8 and 5.4 for oesophageal cancer, respectively. For the cigarette brand smoked in the previous six months, the ORs for >or=10 mg compared with <10 mg were 1.9 for cancer of the oral cavity and pharynx and 1.8 for oesophageal cancer, after allowance for number of cigarettes and duration of smoking. CONCLUSIONS: The present study confirms the direct relationship between the tar yield of cigarettes and upper digestive tract neoplasms, and provides innovative information on lower tar cigarettes, which imply reduced risks compared with higher tar ones. However, significant excess risks were observed even in the lower tar category, thus giving unequivocal indications for stopping smoking as a priority for prevention of upper digestive tract neoplasms. PMID- 12562647 TI - Chemosensitivity and p53-Bax pathway-mediated apoptosis in patients with uterine cervical cancer. AB - OBJECTIVES: To determine whether and how apoptosis through the p53-Bax pathway affects sensitivity to chemotherapy in cervical cancer. MATERIALS AND METHODS: Thirty patients with cervical squamous cell carcinoma, who had human papilloma virus (HPV) and underwent neoadjuvant chemotherapy, were entered in the present study. Tumor specimens were obtained before and after chemotherapy. HPV was detected by polymerase chain reaction. The expression of Ki-67, p53, Bax and Bcl 2 proteins was determined by immunohistochemical staining. Apoptotic cells were identified by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labeling method. RESULTS: Of 30 patients, 18 responded to chemotherapy and 12 did not. The apoptotic index in tumors of responders was significantly higher than in non-responders after chemotherapy. The Ki-67 labeling index (LI) in responders was significantly higher than in non responders before chemotherapy. Patients with tumors >33% of the LI, which was determined by a receiver operating characteristic curve, had a better survival rate. The incidence of p53 protein expression did not differ between responders and non-responders. After chemotherapy, the expression of Bax protein in responders was more frequent and Bcl-2 protein expression was less frequent than in non-responders. CONCLUSIONS: Chemosensitivity in cervical cancer may be associated with apoptosis via the p53-Bax pathway. PMID- 12562648 TI - Activation of the receptor protein tyrosine kinase EphB4 in endometrial hyperplasia and endometrial carcinoma. AB - BACKGROUND: Members of the Eph family of tyrosine kinases have been implicated in embryonic pattern formation and vascular development; however, little is known about their role in the adult organism. We have observed estrogen-dependent EphB4 expression in the normal breast suggesting its implication in the hormone controlled homeostasis of this organ. Since the endometrium is a similarly hormone dependent organ and endometrial carcinoma is thought to result from estrogenic stimulation, we have investigated EphB4 expression in normal human endometrium and during its carcinogenesis. PATIENTS AND METHODS: EphB4 expression was analyzed immunohistochemically in 26 normal endometrium specimens, 15 hyperplasias and 102 endometrioid adenocarcinomas and correlated with clinical and prognostic tumor characteristics. RESULTS: In normal endometrial tissue no EphB4 protein was detected. Strikingly, we observed a drastic increase (P <0.0001) in the number of EphB4 protein-expressing glandular epithelial cells in the majority of hyperplasias and carcinomas. Moreover, we found a statistically highly significant positive correlation between EphB4 expression and post menopausal stage of the patient (P = 0.007). CONCLUSIONS: These findings indicate that in the endometrium, EphB4 is an early indicator of malignant development and, thus, EphB4 may represent a potent tool for diagnosis and therapeutic intervention. PMID- 12562649 TI - Accelerated versus standard cyclophosphamide, epirubicin and 5-fluorouracil or cyclophosphamide, methotrexate and 5-fluorouracil: a randomized phase III trial in locally advanced breast cancer. AB - BACKGROUND: The purpose of this study was to evaluate the impact of a dose-dense primary chemotherapy on pathological response rate (pCR) in patients with locally advanced breast cancer (LABC) treated with combined modality therapy. PATIENTS AND METHODS: Stage IIIA/IIIB patients received three courses of induction chemotherapy (ICT) with cyclophosphamide, epirubicin and 5-fluorouracil (CEF) followed by local therapy (total mastectomy or segmental mastectomy with axillary nodes dissection) and adjuvant chemotherapy (ACT) with three courses of CEF alternated with three courses of cyclophosphamide, methotrexate, 5-fluorouracil (CMF). Patients were randomized to receive ICT and ACT every 3 weeks (arm A, 'standard treatment') or every 2 weeks with granulocyte-macrophage colony stimulating factor (GM-CSF) support (arm B, 'dose-dense treatment'). In both arms radiotherapy was administered after the end of chemotherapy (in selected cases) and patients with hormonal receptor-positive tumors received tamoxifen for 5 years. RESULTS: A total of 150 patients were randomized (77 arm A and 73 arm B) and demographics were well balanced between the two arms. Compliance to treatment was excellent: 95% and 93% of patients in arms A and B, respectively, completed the treatment program with no modification or delay. Median duration of treatment (ICT+local+ACT) was 183 days (range 0-265) in arm A and 139 days (0-226) in arm B. The average relative dose intensity (ARDI) of chemotherapy was 1.3 with a 30% increase in the dose intensity in arm B in comparison with arm A. No difference in clinical [62%; 95% confidence interval (CI) 49% to 73.2%] and pathological response rates to ICT was observed between the two arms. Median follow-up was 5 years (range 1-96 months); median disease-free survivals were 4.8 years in arm A and 4.5 years in arm B. Median overall survival was 7.8 years in standard therapy: this figure has not yet been reached in the dose-dense treatment. CONCLUSIONS: In LABC a dose-dense regimen, while allowing a 30% increase in the dose intensity of chemotherapy, did not provide significant improvement in pathological response rates. However, accelerated chemotherapy reduced the duration of the combined-modality program (6.1 versus 4.6 months) with no additional toxicities. PMID- 12562650 TI - Idoxifene versus tamoxifen: a randomized comparison in postmenopausal patients with metastatic breast cancer. AB - BACKGROUND: More efficacious and safer hormonal agents are needed for breast cancer treatment and prevention. Idoxifene is a novel selective estrogen receptor modulator (SERM) that, in preclinical models, has greater antiestrogenic but lower estrogenic activity than tamoxifen. PATIENTS AND METHODS: Three hundred and twenty-one postmenopausal patients with hormone receptor-positive or -unknown metastatic breast cancer were randomized to receive either tamoxifen or idoxifene as initial endocrine therapy for advanced disease. Data were analyzed based on intention to treat and all the responses were subject to independent review. RESULTS: At the time of a second planned interim analysis, the trial was stopped for economic considerations, not for reasons related to safety or efficacy. Complete data for the 219 patients included in the second interim analysis are fully available and reported here. Median age was 59.1 years for idoxifene patients and 59.9 years for tamoxifen patients. Complete response (CR) plus partial response (PR) rates were as follows: tamoxifen, 9%; idoxifene, 13% (P = 0.39). Clinical benefit rate [CR + PR + stable disease (SD) >or=6 months] was 34.3% for idoxifene and 38.7% for tamoxifen (P = 0.31). Median time to progression and duration of response were 140 days and 151.5 days, respectively, for tamoxifen compared with 166 days and 218 days for idoxifene. None of these endpoints was significantly different for the two drugs, nor was survival. Adverse events (lethal, serious but not lethal and important but not life threatening) were similar in the two arms. CONCLUSIONS: Idoxifene was both active and well tolerated in postmenopausal women with metastatic breast cancer. Idoxifene had similar efficacy and toxicity to tamoxifen in this randomized comparison. PMID- 12562651 TI - Preclinical and clinical evaluation of four gemcitabine plus carboplatin schedules as front-line treatment for stage IV non-small-cell lung cancer. AB - BACKGROUND: To explore the activity and tolerability of gemcitabine (GEM) and carboplatin (CBDCA) in non-small-cell lung cancer (NSCLC) we tested four administration sequences on H460 NSCLC cells, and at the same time performed a randomized phase II trial using analogous schedules. PATIENTS AND METHODS: GEM was given first in two in vitro sequences, and CBDCA first in the other two; interaction was quantified calculating a combination index. Eighty-eight chemotherapy-naive, stage IV NSCLC patients were randomly assigned to receive either: GEM (1000 mg/m(2)) on days 1 and 8 and CBDCA (AUC 5 mg.min/ml) on day 1, 4 h before GEM (arm A); same as arm A except CBDCA given 4 h after GEM (arm B); GEM on days 1 and 8 and CBDCA on day 2 (arm C); GEM on days 2 and 9 and CBDCA on day 1 (arm D). Courses were repeated every 21 days. RESULTS: In the preclinical study, CBDCA given before GEM produced a synergistic cytotoxic effect. Two complete and 29 partial responses occurred in 86 of 88 treated patients (intention-to-treat analysis 35%; 95% confidence interval 25.5% to 46.8%). One- and 2-year survivals were 44% and 11%, respectively. Grade 3/4 thrombocytopenia occurred in 11%; grade 3/4 neutropenia in 17%; and non-hematological toxicity was insignificant. Median survival was 11 months (range 7-18+), but better in patients receiving CBDCA first (arms A and D) (13 versus 9 months) than in patients receiving GEM first (arms B and C). The response was greater (50% versus 31%) in arm A than in the other arms. CONCLUSIONS: The CBDCA/GEM combination is safe and active against stage IV NSCLC. Our preclinical and clinical findings suggest that administration of CBDCA before GEM gives the better outcome. PMID- 12562652 TI - Economic evaluation of antibiotic prophylaxis in small-cell lung cancer patients receiving chemotherapy: an EORTC double-blind placebo-controlled phase III study (08923). AB - BACKGROUND: To determine whether the cost of prophylactic antibiotics during chemotherapy is offset by cost savings due to a decreased incidence of febrile leukopenia (FL). PATIENTS AND METHODS: Small-cell lung cancer (SCLC) patients were randomised to standard or intensified chemotherapy with granulocyte colony stimulating factor to assess the impact on survival (n = 244). In addition, patients were randomised to prophylactic ciprofloxacin and roxithromycin or placebo to assess the impact on FL (n = 161). The economic evaluation examined the costs and effects of patients taking antibiotics versus placebo. Medical resource utilisation was documented prospectively, including 33 patients from one centre in The Netherlands (NL) and 49 patients from one centre in Germany (GE). The evaluation takes the perspective of the health insurance systems and of the hospitals. Sensitivity analyses were performed. RESULTS: In the main trial, prophylactic antibiotics reduced the incidence of FL, hospitalisation due to FL and use of therapeutic antibiotics by 50%. In GE, the incidence of FL was not reduced by prophylaxis. This resulted in an average cost difference of only 35 Euros [95% confidence interval (CI) (-)1.713-2.263] in favour of prophylaxis (not significant). In NL, prophylaxis reduced the incidence of FL by nearly 50%, comparable with the results of the main trial, resulting in a cost difference of 2706 Euros [95% CI 810-5948], demonstrating savings in favour of prophylactic antibiotics of nearly 45%. Sensitivity analyses indicate that with an efficacy of prophylaxis of 50%, and with expected costs of antibiotic prophylaxis of 500 Euros or less, cost savings will incur over a broad range of baseline risks for FL; that is, a risk >10-20% for FL per cycle. CONCLUSIONS: Giving oral prophylactic antibiotics to SCLC patients undergoing chemotherapy is the dominant strategy in both GE and NL, demonstrating both cost-savings and superior efficacy. The sensitivity analyses demonstrate that, due to the efficacy of prophylactic antibiotics and their low unit cost, cost savings will incur over a broad range of baseline risks for FL. We recommend the use of prophylactic antibiotics in patients at risk for FL during chemotherapy. PMID- 12562653 TI - A randomised multicentre trial of modified CHOP versus MCOP in patients aged 65 years and over with aggressive non-Hodgkin's lymphoma. AB - BACKGROUND: The aim of this study was to determine in a randomised trial whether there is any significant difference in toxicity between modified CHOP and MCOP chemotherapy in elderly patients with aggressive non-Hodgkin's lymphoma (NHL) and to determine whether this reduced dose chemotherapy can be administered with full dose intensity, low toxicity and produce acceptable survival. PATIENTS AND METHODS: Between 1993 and 2000, 155 eligible patients were randomised into this trial mainly from three centres (Nottingham, Birmingham and Leeds, UK). The patients were newly diagnosed with aggressive NHL and had a median age of 74 years (range 65-91 years). Ninety-six patients (62%) had bulky stage I or II disease; 59 patients (38%) had either stage III or IV disease; 77% had one or more extranodal sites involved at presentation; and 31% showed B symptoms. Seventy-seven patients were randomised to receive six cycles of modified CHOP (cyclophosphamide 600 mg/m(2) i.v., doxorubicin 30 mg/m(2) i.v., vincristine 1 mg i.v. all on day 1 with prednisolone 20 mg bd for days 1-5) every 21 days and 78 patients to MCOP (mitozantrone 10 mg/m(2) i.v. substituted for doxorubicin). Growth factors were not used routinely. After completion of chemotherapy, 39 patients received involved field radiotherapy (35-40 Gy) in 20 fractions. RESULTS: One hundred and one patients (65%) completed all six cycles of chemotherapy. The median course dose intensity was 97%. The median follow-up for 53 surviving patients was 51 months. The median survival was 19 months (95% confidence interval 10-36 months) with an actuarial survival of 47% at 2 years and 42% at 3 years (CHOP versus MCOP, P = 0.79). There was no significant difference in any of the toxicities experienced with either CHOP or MCOP, except for white cell count (46 patients on MCOP and 27 patients on CHOP had grade 3 or 4 toxicity, P = 0.002) and red cell transfusion (37 patients, MCOP; 17 patients, CHOP; P = 0.001). Grade 3 or 4 neutropenia was documented in 75 patients (50%). One patient died from toxicity whilst in remission and seven patients died with septicaemia and persistent NHL. CONCLUSION: This multicentre randomised trial provides further information on the dose intensity achievable with CHOP or MCOP regimens in elderly patients (median age 74 years) with aggressive NHL. These dose-reduced regimens can be given with nearly 100% dose intensity with 65% of patients completing all the treatment. Survival is comparable to that observed with the more intensive regimens given in this age group. PMID- 12562654 TI - Primary systemic treatment of advanced Hodgkin's disease with EVA (etoposide, vinblastine, doxorubicin): 10-year follow-up. AB - BACKGROUND: The most commonly used regimen for the treatment of advanced Hodgkin's disease (HD) is ABVD (doxorubicin, bleomycin, vinblastine and dacarbazine). Two of these components, bleomycin and dacarbazine, have defined toxicities such as pulmonary fibrosis and nausea/vomiting, and also uncertain single-drug activity. The EVA regimen (etoposide, vinblastine, doxorubicin) is an attempt to substitute a known active agent, etoposide, for bleomycin and dacarbazine. PATIENTS AND METHODS: A series of 51 patients with advanced HD without prior systemic therapy were treated. The series included 12 stage II patients with bulky (>10 cm) mediastinal tumors, 10 of whom received complementary radiation therapy. The remaining patients received EVA only. Response, duration of response, survival, toxicity and the efficacy of salvage therapy were evaluated in all patients. The median follow-up time was 111 months and permitted an assessment of the long-term effects of treatment and natural history of a cohort of treated patients. RESULTS: EVA achieved a complete response (or clinical complete response) in 48/51 patients (94%). Of these 48 responders, 16 relapsed in a median of 11 months (range 3-48 months). In follow up, 32/51 patients had no evidence of relapsed HD, although three died from other causes (two from vascular events and one from large cell lymphoma), resulting in progression-free survival for the entire group of 57% at 111 months. Eight of the 16 were alive and free from disease at follow-up at 111 months. In the entire series, only seven patients (14%) died of HD. 37 patients (73%) continued free from disease. There was no pulmonary toxicity. CONCLUSIONS: The EVA regimen appears to have an overall survival (OS) outcome comparable to ABVD, but without the lung toxicity. The high salvage rate of second-line therapy, in most instances at conventional dosage, suggests an absence of cross-resistance to alkylating agents in patients treated with EVA. PMID- 12562655 TI - Optimum trephine length in the assessment of bone marrow involvement in patients with diffuse large cell lymphoma. AB - BACKGROUND: The National Cancer Institute has recommended a bone marrow biopsy length of >/=20 mm for the staging and surveillance of patients with non Hodgkin's lymphoma. However, there are few published data to support this recommendation, particularly the role of examining multiple levels. PATIENTS AND METHODS: Bone marrow biopsies from 172 patients with newly diagnosed diffuse large cell lymphoma (DLCL) entered in two consecutive trials of the Australasian Leukaemia and Lymphoma Group were analysed. The original haematoxylin and eosin stained trephine biopsy and two or more deeper sections cut at 0.1-0.2 mm intervals were assessed with respect to the morphology, extent and pattern of lymphomatous involvement. The rate of positive diagnosis was correlated with the length of the biopsy specimen and the number of sections examined. RESULTS: Forty seven biopsies (27%) demonstrated marrow involvement on examination of a mean of four trephine biopsy sections. The rate of positivity increased with the examination of multiple levels and correlated with increasing trephine length but was not dependent on the number of sites sampled. Twenty per cent of biopsies <20 mm in length were positive for lymphoma; this increased to 35% for biopsies >/=20 mm (P = 0.023). CONCLUSIONS: Morphological bone marrow involvement in DLCL is optimally demonstrated by a 20-mm long trephine biopsy from a single site which is examined at multiple levels (four or more). This obviates the need for bilateral sampling, thereby reducing patient morbidity from the procedure. This study provides evidence to support the National Cancer Institute recommendations regarding trephine biopsy in the staging of DLCL, providing multiple levels are examined. PMID- 12562656 TI - Early cardiotoxicity of the CHOP regimen in aggressive non-Hodgkin's lymphoma. AB - BACKGROUND: To determine the incidence of early cardiotoxicity induced by the CHOP regimen in patients with aggressive non-Hodgkin's lymphoma (NHL) and to identify associated risk factors. PATIENTS AND METHODS: A retrospective analysis included 135 consecutive patients who had been treated with the CHOP (cyclophosphamide, doxorubicin, vincristin, prednisone) regimen as first-line therapy between 1994 and 2000. The cardiac evaluation was based on a determination of the resting left ventricular ejection function (LVEF) by gated blood-pool imaging. Cardiotoxicity was defined as a significant decrease in LVEF or clinical evidence of congestive heart failure (CHF). RESULTS: Twenty-seven (20%) patients developed a cardiac event within 1 year of treatment. Among these, 14 patients had clinical signs of CHF. Three patients died suddenly from presumed cardiac causes. In multivariate analysis, a cumulative dose of doxorubicin >200 mg/m(2) [odds ratio (OR) = 4.2, P = 0.005)] and age over 50 years (OR = 2.9, P = 0.03) appeared to be significant risk factors. CONCLUSION: Early clinical and subclinical cardiotoxicity was frequent in patients receiving the CHOP regimen. The threshold of the cumulative dose of doxorubicin appeared to be low: at doses >200 mg/m(2), 27% of patients had cardiac events. Elderly patients appeared to be at higher risk. The development of cardioprotective strategies or alternative treatments are mandatory for aggressive NHL patients. PMID- 12562657 TI - The effect of Epstein-Barr virus status on outcome in age- and sex-defined subgroups of patients with advanced Hodgkin's disease. AB - BACKGROUND: Conflicting data on the effect of the Epstein-Barr virus (EBV) on outcome in Hodgkin's disease (HD) might be due to the heterogeneous nature of this disease. In this study we have investigated whether the effect of EBV status on outcome is different between aetiologically defined age groups (15-34, 35-44, 45+ years) and also between males and females. PATIENTS AND METHODS: Paraffin embedded sections from 273 patients with advanced HD from two related clinical trials were analysed for the presence of EBV using in situ hybridisation. RESULTS: EBV was detected in 78 (29%) of cases. For all patients, after a median follow-up of 5 years, there were no significant differences in survival by EBV status although there was a trend towards longer failure-free survival times for EBV-positive patients. Multivariate analyses suggested that EBV and sex, when in combination, were prognostic factors for failure-free survival (P = 0.06 for both). For subgroups, the effect of EBV on failure-free survival was significant for males and 15-34 years age group (P = 0.05 and P = 0.03, respectively). CONCLUSION: This study suggests that with a median follow-up of 5 years, EBV status does not affect survival but being EBV-positive may be beneficial in terms of failure-free survival, particularly for males and younger adults. PMID- 12562658 TI - A randomized double-blind trial to compare the clinical efficacy of granisetron with metoclopramide, both combined with dexamethasone in the prophylaxis of chemotherapy-induced delayed emesis. AB - BACKGROUND: The prophylactic use of 5-HT(3) receptor antagonists (setrons), after the first 24 h (acute phase) of exposure to emetic chemotherapy, to decrease the incidence of 'delayed phase' emesis increases costs. We designed a study to evaluate the efficacy of a setron (granisetron) in the delayed phase, compared with metoclopramide, each combined with a corticosteroid. PATIENTS AND METHODS: Patients on their first course of single-day emetic chemotherapy (cisplatin, carboplatin, doxorubicin, cyclophosphamide and others) received granisetron 2 mg p.o. and dexamethasone 8 mg p.o. on day 1, followed for 5 days by dexamethasone 4 mg p.o. od combined with either metoclopramide 20 mg p.o. tds or granisetron 1 mg bd in a double-blinded double-dummy protocol. Patients evaluated the results using a diary card. Randomization was stratified by institution, sex, emetic chemotherapy naive versus previous, alcohol consumption and platinum versus non platinum regimen. RESULTS: 131 evaluable patients received granisetron in the delayed phase, and 127 received metoclopramide. Control of acute emesis in both arms was similar (86% granisetron; 85% metoclopramide). The 35 patients experiencing acute emesis had poor control in the delayed phase, with only four granisetron and three metoclopramide patients having no or mild nausea and no vomiting. CONCLUSIONS: In daily practice, a combination of oral dexamethasone and oral granisetron achieves an extremely high control of acute emesis (86% protection). Our data suggest that routine prescription of setrons for delayed phase control is not advisable as it increases costs without any benefit for the majority of patients. Delayed emesis in the rare patients with acute phase emesis remains an unsolved problem. PMID- 12562659 TI - Phase I study of combined radiation, hyperthermia and intra-arterial carboplatin for local recurrence of cervical cancer. AB - BACKGROUND: Patients with cervical cancer who develop pelvic recurrence after primary surgery are usually treated with radiation-based therapy. However, their prognoses are dismal. We conducted a phase I study of combined radiation, hyperthermia and intra-arterial (IA) carboplatin for local recurrence of cervical cancer. PATIENTS AND METHODS: Patients with local recurrence of cervical cancer without extrapelvic recurrence were included in this study. Carboplatin was given as a 5-min IA infusion without hydration just before pelvic radiation every day. External pelvic irradiation (1.8 Gy/day for 28 days) was performed according to local standard schedules. After 20 Gy had been administered, hyperthermia was performed once a week with a radio frequency heating system for four cycles. RESULTS: Fifteen patients were entered through the four dose levels of carboplatin. The maximum tolerated dose was determined to be 25 mg/m(2 )and the dose-limiting toxicities were leukocytopenia, neutrocytopenia and diarrhea. Grade 3/4 leukocytopenia and diarrhea were observed in nine (60%) and three (20%) of 15 patients. Tumor responses included five complete responses and nine partial responses, and the overall response rate was 93.3% (14 of 15) (95% confidence interval 59.4% to 100%). Tumor reductions were observed only at 20 Gy in 10 cases of 14 responders (71.4%). CONCLUSION: The combination therapy of radiation, hyperthermia and IA carboplatin is safe and well-tolerated for locally recurrent cervical cancer. PMID- 12562660 TI - A dose-escalation and pharmacokinetic study of gemcitabine and oxaliplatin in patients with advanced solid tumors. AB - BACKGROUND: Gemcitabine and oxaliplatin have broad antineoplastic activity and favorable toxicity. We conducted a phase I study to determine the maximum tolerated doses (MTDs) and dose-limiting toxicities (DLTs) of the combination in patients with advanced solid tumors. PATIENTS AND METHODS: Sixty-eight patients with advanced stage solid tumors were enrolled. Treatment was first-line for 35% of patients, second-line for 27%, and third-line for 38%. Gemcitabine was administered at escalating doses of 1000-2000 mg/m(2) as a 30-min intravenous (i.v.) infusion on days 1 and 8 and oxaliplatin at 60-130 mg/m(2) as a 4-h i.v. infusion on day 8 every 21 days without growth factor support. RESULTS: The MTD was defined at gemcitabine 1800 mg/m(2) on days 1 and 8 and oxaliplatin 130 mg/m(2) on day 8. Twelve dose levels were evaluated and DLTs occurring during the first cycle consisted of grade 4 neutropenia, grade 3 asthenia or mucositis and grade 1-3 neutropenia or thrombocytopenia resulting in treatment delays. A total of 266 cycles were administered with only one episode of febrile neutropenia and no toxic deaths. Seven (3%) and 26 (10%) cycles were complicated by grade 4 and 3 neutropenia, respectively, three (1%) and 13 (5%) by grade 4 and 3 thrombocytopenia, and eight (3%) by grade 3 anemia. The most common non hematological toxicity was grade 2/3 asthenia observed in 23% of cycles. Responses were observed in patients with a variety of epithelial neoplasms. The pharmacokinetic study revealed no significant interaction between the two drugs. CONCLUSIONS: The combination of gemcitabine and oxaliplatin has excellent tolerability and promising activity in patients with advanced solid tumors. As the MTD exceeds the recommended single-agent dose for gemcitabine, and a dose response effect has not been established, we recommend using both drugs at full doses, e.g. gemcitabine 1200-1400 mg/m(2) on days 1 and 8 and oxaliplatin 130 mg/m(2) on day 8 for further phase II studies. PMID- 12562661 TI - Cancer prevalence in Central Europe: the EUROPREVAL Study. AB - BACKGROUND: Information on cancer prevalence is either absent or largely unavailable for central European countries. MATERIALS AND METHODS: Austria, Germany, The Netherlands, Poland, Slovakia, Slovenia and Switzerland cover a population of 13 million inhabitants. Cancer registries in these countries supplied incidence and survival data for 465 000 cases of cancer. The prevalence of stomach, colon, rectum, lung, breast, cervix uteri, corpus uteri and prostate cancer, as well as skin melanoma, Hodgkin's disease, leukaemia and all malignant neoplasms combined was estimated for the end of 1992. RESULTS: A large heterogeneity was observed within central European countries. For all cancers combined, estimates ranged from 730 per 100 000 in Poland (men) to 3350 per 100 000 in Germany (women). Overall cancer prevalence was the highest in Germany and Switzerland, and the lowest in Poland and Slovenia. In Slovakia, prevalence was higher than average for men and lower than average for women. This was observed for almost all ages. As shown by incidence data, breast cancer was the most frequent malignancy among women in all countries. Among men, prostate cancer was the leading malignancy in Germany, Austria and Switzerland, and lung cancer was the major cancer in Slovenia, Slovakia and Poland. The Netherlands had a high prevalence of both prostate and lung cancer. Time-related magnitude of prevalence within each country and the variability of such proportions across the countries has been estimated and cancer prevalence is given by time since diagnosis (1 year, 1-5 years, 5-10 years, >10 years) for each site. The weight of 1-year prevalence (248 per 100 000 among men and 253 per 100 000 among women) was <15% of total prevalence. Prevalent cases between 1 and 5 years since diagnosis represented between 22% and 34% of the total prevalence. Prevalent cases diagnosed from 5 to 10 years before (335 per 100 000 for men and 505 per 100 000 for women) represented between 17% and 23% of prevalent cancers. Finally, long term cancer prevalence (diagnosed >10 years before), reflecting long-term survival, and number of people considered as cured from cancer were 490 per 100 000 for men and 1028 per 100 000 for women, with a range between 26% (The Netherlands, men) and 50% (Slovakia, women). CONCLUSION: It is clear from observing countries in Central Europe, that high cancer prevalence is associated with well-developed economies. This burden of cancer could be interpreted as a paradoxical effect of better treatments and thereby survival. It could also be taken as a sign for not being satisfied with the advances in treating patients diagnosed with cancer, and for supporting more primary prevention. PMID- 12562662 TI - Cancer mortality in Ireland, 1926-1995. AB - BACKGROUND: Investigation of long time series of cancer data can still be very useful in helping to identify Cancer Control priorities and achievements. Since the partition of Ireland into the independent Republic of Ireland and Northern Ireland, which remained part of the United Kingdom, cancer mortality data have been published in an essentially similar format in both countries. The information presented here will contribute to providing a basis for the collaborative Cancer Research programme initiated recently. PATIENTS AND METHODS: Cancer mortality data have been assembled and analysed separately for the Republic of Ireland and Northern Ireland: the data have then been combined to present mortality rates for the whole of Ireland, covering the period from 1926 to 1995. Several rubrics had to be aggregated to provide data continuously over the time span (e.g. colon and rectum and cervix and body of the uterus). When data were only available in 10-year classes of age, the EM algorithm was employed to obtain 5-year age-specific rates. All rates presented are age-standardised, employing the World Standard Population. RESULTS: In women, the death rate from all neoplasms combined increased very slightly from 117 per 100 000 in 1946-1950 to 120 per 100 000 in 1991-1995. In men, the death rate increased from 127 per 100 000 to 172 per 100 000 over the same time period. The overall cancer death rate in Ireland is currently similar to the European average in men, although in women it is among the top fifth of national cancer mortality rates in European countries. While cancer is a major cause of death in Ireland, there is no evidence of an evolving epidemic building up: the death rates from most forms of cancer are declining towards the end of the time period considered. CONCLUSIONS: As demonstrated by falling death rates from Hodgkin's disease and testicular cancer, major treatment advances appear to have been incorporated effectively into clinical practice in Ireland. Progress is apparent in tobacco control and further initiatives in this area must be undertaken since tobacco appears to be the only major new carcinogen introduced recently into the Irish environment during the period covered by this study. Effective population-based screening programmes for cervix and breast cancer and, more controversially, consideration of a National Prostate Cancer Screening programme, offer scope for further improvement in mortality. Examination of this long time series of mortality data from Ireland provides information about the evolving cancer pattern and provides the necessary background to evaluate the impact of the cross-border cancer research activities now being launched. PMID- 12562663 TI - Taxane-induced nail changes: incidence, clinical presentation and outcome. AB - The clinical characteristics of nail changes in seven patients receiving taxane containing chemotherapy are described. They include nail pigmentation, subungual hematoma, Beau's lines and onycholysis and subungual suppuration. The incidence of such changes (ranging from 0% to 44%) is reviewed from a Medline search of the literature. PMID- 12562664 TI - Amelioration of docetaxel/cisplatin induced polyneuropathy by alpha-lipoic acid. PMID- 12562665 TI - Fatal herpes simplex virus hepatitis complicating chemotherapy with weekly docetaxel. PMID- 12562666 TI - Lethal 5-fluorouracil toxicity associated with a novel mutation in the dihydropyrimidine dehydrogenase gene. PMID- 12562667 TI - Prognostic markers for survival after high-dose chemotherapy with autologous stem cell transplantation for breast cancer. PMID- 12562668 TI - Debate on colorectal cancer screening by faecal occult blood. PMID- 12562669 TI - Training-induced changes in skeletal muscle Na+-K+ pump number and isoform expression in rats with chronic heart failure. AB - The mechanisms responsible for the decrements in exercise performance in chronic heart failure (CHF) remain poorly understood, but it has been suggested that sarcolemmal alterations could contribute to the early onset of muscular fatigue. Previously, our laboratory demonstrated that the maximal number of ouabain binding sites (B(max)) is reduced in the skeletal muscle of rats with CHF (Musch TI, Wolfram S, Hageman KS, and Pickar JG. J Appl Physiol 92: 2326-2334, 2002). These reductions may coincide with changes in the Na(+)-K(+)-ATPase isoform (alpha and beta) expression. In the present study, we tested the hypothesis that reductions in B(max) would coincide with alterations in the alpha- and beta subunit expression of the sarcolemmal Na(+)-K(+)-ATPase of rats with CHF. Moreover, we tested the hypothesis that exercise training would increase B(max) along with producing significant changes in alpha- and beta-subunit expression. Rats underwent a sham operation (sham; n = 10) or a surgically induced myocardial infarction followed by random assignment to either a control (MI; n = 16) or exercise training group (MI-T; n = 16). The MI-T rats performed exercise training (ET) for 6-8 wk. Hemodynamic indexes demonstrated that MI and MI-T rats suffered from severe left ventricular dysfunction and congestive CHF. Maximal oxygen uptake (Vo(2 max)) and endurance capacity (run time to fatigue) were reduced in MI rats compared with sham. B(max) in the soleus and plantaris muscles and the expression of the alpha(2)-isoform of the Na(+)-K(+)-ATPase in the red portion of the gastrocnemius (gastrocnemius(red)) muscle were reduced in MI rats. After ET, Vo(2 max) and run time to fatigue were increased in the MI-T group of rats. This coincided with increases in soleus and plantaris B(max) and the expression of the alpha(2)-isoform in the gastrocnemius(red) muscle. In addition, the expression of the beta(2)-isoform of the gastrocnemius(red) muscle was increased in the MI-T rats compared with their sedentary counterparts. This study demonstrates that CHF induced alterations in skeletal muscle Na(+)-K(+)-ATPase, including B(max) and isoform expression, can be partially reversed by ET. PMID- 12562670 TI - Enhancement of whole body glucose uptake during and after human skeletal muscle low-frequency electrical stimulation. AB - There is considerable evidence to suggest that electrical stimulation (ES) activates glucose uptake in rodent skeletal muscle. It is, however, unknown whether ES can lead to similar metabolic enhancement in humans. We employed low frequency ES through surface electrodes placed over motor points of quadriceps femoris muscles. In male subjects lying in the supine position, the highest oxygen uptake was obtained by a stimulation pattern with 0.2-ms biphasic square pulses at 20 Hz and a 1-s on-off duty cycle. Oxygen uptake was increased by approximately twofold throughout the 20-min stimulation period and returned to baseline immediately after stimulation. Concurrent elevation of the respiratory exchange ratio and blood lactate concentration indicated anaerobic glycogen breakdown and utilization during ES. Whole body glucose uptake determined by the glucose disposal rate during euglycemic clamp was acutely increased by 2.5 mg. kg(-1). min(-1) in response to ES and, moreover, remained elevated by 3-4 mg. kg( 1). min(-1) for at least 90 min after cessation of stimulation. Thus the stimulatory effect of ES on whole body glucose uptake persisted not only during, but also after, stimulation. Low-frequency ES may become a useful therapeutic approach to activate energy and glucose metabolism in humans. PMID- 12562671 TI - Density-dependent reduction of nitric oxide diffusing capacity after pneumonectomy. AB - Airway lengthening after pneumonectomy (PNX) may increase diffusive resistance to gas mixing (1/D(G)); the effect is accentuated by increasing acinar gas density but is difficult to detect from lung CO-diffusing capacity (Dl(CO)). Because lung NO-diffusing capacity (Dl(NO)) is three- to fivefold that of Dl(CO), whereas 1/D(G) for NO and CO are similar, we hypothesized that a density-dependent fractional reduction would be greater for Dl(NO) than for Dl(CO). We measured Dl(NO) and Dl(CO) at two tidal volumes (Vt) and with three background gases [helium (He), nitrogen (N(2)), and sulfur hexafluoride (SF(6))] in immature dogs 3 and 9 mo after right PNX (5 and 11 mo of age). At maturity (11 mo), background gas density had no effect on Dl(NO), Dl(CO), or Dl(NO)-to-Dl(CO) ratio in sham controls. In PNX animals, Dl(NO) declined 25-50% in SF(6) relative to He and N(2), and Dl(NO)/Dl(CO) declined approximately 50% in SF(6) relative to He at a Vt of 15 ml/kg, consistent with a significant 1/D(G). At 5 mo of age, Dl(NO)/Dl(CO) declined 25-45% in SF(6) relative to He and N(2) in both groups, but Dl(CO) increased paradoxically in SF(6) relative to N(2) or He by 20-60%. Findings suggest that SF(6), besides increasing 1/D(G), may redistribute ventilation and/or enhance acinar penetration of the convective front. PMID- 12562672 TI - Changes in actomyosin ATP consumption rate in rat diaphragm muscle fibers during postnatal development. AB - Early postnatal development of rat diaphragm muscle (Dia(m)) is marked by dramatic transitions in myosin heavy chain (MHC) isoform expression. We hypothesized that the transition from the neonatal isoform of MHC (MHC(Neo)) to adult fast MHC isoform expression in Dia(m) fibers is accompanied by an increase in both the maximum velocity of the actomyosin ATPase reaction (V(max) ATPase) and the ATP consumption rate during maximum isometric activation (ATP(iso)). Rat Dia(m) fibers were evaluated at postnatal days 0, 14, and 28 and in adults (day 84). Across all ages, V(max) ATPase of fibers was significantly higher than ATP(iso). The reserve capacity for ATP consumption [1 - (ratio of ATP(iso) to V(max) ATP(ase))] was remarkably constant ( approximately 55-60%) across age groups, although at day 28 and in adults the reserve capacity for ATP consumption was slightly higher for fibers expressing MHC(Slow) compared with fast MHC isoforms. At day 28 and in adults, both V(max) ATPase and ATP(iso) were lower in fibers expressing MHC(Slow) followed in rank order by fibers expressing MHC(2A), MHC(2X), and MHC(2B). For fibers expressing MHC(Neo), V(max) ATPase, and ATP(iso) were comparable to values for adult fibers expressing MHC(Slow) but significantly lower than values for fibers expressing fast MHC isoforms. We conclude that postnatal transitions from MHC(Neo) to adult fast MHC isoform expression in Dia(m) fibers are associated with corresponding but disproportionate changes in V(max) ATPase and ATP(iso). PMID- 12562673 TI - Measurement of mitochondrial DNA synthesis in vivo using a stable isotope-mass spectrometric technique. AB - We describe here a new stable isotope-mass spectrometric technique for measuring mitochondrial DNA (mtDNA) synthesis. Growing (2-4 mo old) and weight-stable (8-10 mo old) Sprague-Dawley rats were primed with (2)H(2)O (deuterated water) to 2.0 2.5% body water enrichment, via intraperitoneal injection, and then given 4% (2)H(2)O in drinking water for 3-11 wk. Mitochondria were isolated from cardiac and hindlimb muscle, and mtDNA was isolated and enzymatically hydrolyzed to deoxyribonucleosides. PCR confirmed the absence of nuclear DNA contamination. The isotopic enrichment of the deoxyribose moiety of deoxyadenosine was determined by GC-MS analysis, and percent new mtDNA was calculated by comparison to genomic DNA enrichments in a tissue with nearly complete turnover (bone marrow). Initial label incorporation into deoxyadenosine of mtDNA was linear, and turnover of mtDNA was observed in nongrowing adult female rats (1.1-1.3% new mtDNA per day in cardiac and skeletal muscle). Die-away curves of mtDNA after discontinuing (2)H(2)O administration gave a similar turnover rate constant. Human subjects were also given (2)H(2)O for up to 6 wk, and mitochondria from platelets were isolated. Incubation with DNase removed any contaminating genomic DNA; platelet mtDNA exhibited linear incorporation from (2)H(2)O and reached plateau values identical to those in genomic DNA from fully turned over cells (circulating monocytes). In conclusion, replication of mtDNA can be directly measured in vivo in rodents and humans without the use of radioactivity. Use of this technique may allow improved understanding of the regulation of mitochondrial biogenesis in health and disease. PMID- 12562674 TI - Measurement of nutritional status in simulated microgravity by bioelectrical impedance spectroscopy. AB - The potential of bioelectrical impedance spectroscopy (BIS) for assessing nutritional status in spaceflight was tested in two head-down-tilt bed-rest studies. BIS-predicted extracellular water (ECW), intracellular water (ICW), and total body water (TBW) measured using knee-elbow electrode placement were compared with deuterium and bromide dilution (DIL) volumes in healthy, 19- to 45 yr-old subjects. BIS was accurate during 44 h of head-down tilt with mean differences (BIS - DIL) of 0-0.1 kg for ECW, 0.3-0.5 for ICW, and 0.4-0.6 kg for TBW (n = 28). At 44 h, BIS followed the within-individual change in body water compartments with a relative prediction error (standard error of the estimate/baseline volume) of 2.0-3.6% of water space. In the second study, BIS did not detect an acute decrease (-1.41 +/- 0.91 kg) in ICW secondary to 48 h of a protein-free, 800 kcal/day diet (n = 18). BIS's insensitivity to ICW losses may be because they were predominantly (65%) localized to the trunk and/or because there was a general failure of BIS to measure ICW independently of ECW and TBW. BIS may have potential for measuring nutritional status during spaceflight, but its limitations in precision and insensitivity to acute ICW changes warrant further validation studies. PMID- 12562675 TI - Assessment of exhaled nitric oxide kinetics in healthy infants. AB - Exhaled nitric oxide (Fe(NO)) measurements provide a noninvasive approach to the evaluation of airway inflammation. Flow-independent NO exchange parameters [airway NO transfer factor (D(NO)) and airway wall NO concentration (Cw(NO))] can be estimated from Fe(NO) measurements at low flows and may elucidate mechanisms of disturbances in NO exchange. We measured Fe(NO) in sedated infants by using an adaptation of a raised lung volume rapid thoracic compression technique that creates forced expiration through a mass-flow controller that lasts 5-10 s, at a constant preset flow. We measured Fe(NO) at expired flows of 50, 25, and 15 ml/s in five healthy infants (7-31 mo). Median Fe(NO) increased [24, 40, and 60 parts per billion (ppb)] with decreasing expiratory flows (50, 25, and 15 ml/s). Group median (range) for D(NO) and Cw(NO) were 12.7 (3.2-37) x 10(-3) nl. s(-1). ppb( 1) and 108.9 (49-385) ppb, respectively, similar to values reported in healthy adults. Exhaled NO is flow dependent; flow-independent parameters of exhaled NO kinetics can be assessed in infants and are similar to values described in adults. PMID- 12562676 TI - Chronic O2 exposure enhances vascular and airway smooth muscle contraction in the newborn but not adult rat. AB - Neonatal rats exposed to 60% O(2) for 14 days develop lung changes compatible with human bronchopulmonary dysplasia and pulmonary hypertension. Our aim was to evaluate and compare the newborn and adult rat pulmonary vascular and airway smooth muscle force generation and relaxation potential after exposure to 60% O(2) for 14 days. Vascular and airway intrapulmonary rings 100 microm in diameter were mounted on a myograph and bathed in Krebs-Henseleit solution bubbled with air- 6% CO(2) at 37 degrees C. Significant age-dependent changes in intrapulmonary arteries and their neighboring airway muscle properties were observed. Whereas hyperoxia enhanced force in neonatal vascular and airway muscle, the opposite was seen in adult samples. No changes in endothelium dependent vascular relaxation were observed at either age, but the dose response to an endothelium-independent NO donor was altered. In the newborn experimental animals, the relaxation was reduced, whereas, in their adult counterparts, it was enhanced. After O(2) exposure, the bronchial muscle relaxation response to epithelium-dependent and -independent stimulation was not altered in either age group, whereas the epithelium-dependent response was decreased only in the adult. The antioxidant Trolox, or an endothelin-A and -B receptor antagonist, reversed the vascular and airway muscle's hyperoxia-induced changes. We conclude that chronic O(2) exposure in the newborn rat results in enhanced lung vascular and airway muscle contraction potential via a mechanism involving reactive oxygen species and the endothelin pathway. The present findings also suggest that the newborn is more susceptible to airway hyperresponsiveness after chronic O(2) exposure. PMID- 12562677 TI - Preexercise hypervolemia does not affect arterial hypoxemia in Thoroughbreds performing short-term high-intensity exercise. AB - It is reported that preexercise hyperhydration caused arterial O(2) tension of horses performing submaximal exercise to decrease further by 15 Torr (Sosa-Leon L, Hodgson DR, Evans DL, Ray SP, Carlson GP, and Rose RJ. Equine Vet J Suppl 34: 425-429, 2002). Because hydration status is important to optimal athletic performance and thermoregulation during exercise, the present study examined whether preexercise induction of hypervolemia would similarly accentuate the arterial hypoxemia in Thoroughbreds performing short-term high-intensity exercise. Two sets of experiments (namely, control and hypervolemia studies) were carried out on seven healthy, exercise-trained Thoroughbred horses in random order, 7 days apart. In resting horses, an 18.0 +/- 1.8% increase in plasma volume was induced with NaCl (0.30-0.45 g/kg dissolved in 1,500 ml H(2)O) administered via a nasogastric tube, 285-290 min preexercise. Blood-gas and pH measurements as well as concentrations of plasma protein, hemoglobin, and blood lactate were determined at rest and during incremental exercise leading to maximal exertion (14 m/s on a 3.5% uphill grade) that induced pulmonary hemorrhage in all horses in both treatments. In both treatments, significant arterial hypoxemia, desaturation of hemoglobin, hypercapnia, acidosis, and hyperthermia developed during maximal exercise, but statistically significant differences between treatments were not found. Thus preexercise 18% expansion of plasma volume failed to significantly affect the development and/or severity of arterial hypoxemia in Thoroughbreds performing maximal exercise. Although blood lactate concentration and arterial pH were unaffected, hemodilution caused in this manner resulted in a significant (P < 0.01) attenuation of the exercise induced expansion of the arterial-to-mixed venous blood O(2) content gradient. PMID- 12562678 TI - Genetic Models in Applied Physiology. Differential role of nitric oxide synthase isoforms in fever of different etiologies: studies using Nos gene-deficient mice. AB - Male C57BL/6J mice deficient in nitric oxide synthase (NOS) genes (knockout) and control (wild-type) mice were implanted intra-abdominally with battery-operated miniature biotelemeters (model VMFH MiniMitter, Sunriver, OR) to monitor changes in body temperature. Intravenous injection of lipopolysaccharide (LPS; 50 microg/kg) was used to trigger fever in response to systemic inflammation in mice. To induce a febrile response to localized inflammation, the mice were injected subcutaneously with pure turpentine oil (30 microl/animal) into the left hindlimb. Oral administration (gavage) of N(G)-monomethyl-l-arginine (l-NMMA) for 3 days (80 mg. kg(-1). day(-1) in corn oil) before injection of pyrogens was used to inhibit all three NOSs (N(G)-monomethyl-d-arginine acetate salt and corn oil were used as control). In normal male C57BL/6J mice, l-NMMA inhibited the LPS induced fever by approximately 60%, whereas it augmented fever by approximately 65% in mice injected with turpentine. Challenging the respective NOS knockout mice with LPS and with l-NMMA revealed that inducible NOS and neuronal NOS isoforms are responsible for the induction of fever to LPS, whereas endothelial NOS (eNOS) is not involved. In contrast, none of the NOS isoforms appeared to trigger fever to turpentine. Inhibition of eNOS, however, exacerbates fever in mice treated with l-NMMA and turpentine, indicating that eNOS participates in the antipyretic mechanism. These data support the hypothesis that nitric oxide is a regulator of fever. Its action differs, however, depending on the pyrogen used and the NOS isoform. PMID- 12562679 TI - Biophysical basis for inner ear decompression sickness. AB - Isolated inner ear decompression sickness (DCS) is recognized in deep diving involving breathing of helium-oxygen mixtures, particularly when breathing gas is switched to a nitrogen-rich mixture during decompression. The biophysical basis for this selective vulnerability of the inner ear to DCS has not been established. A compartmental model of inert gas kinetics in the human inner ear was constructed from anatomical and physiological parameters described in the literature and used to simulate inert gas tensions in the inner ear during deep dives and breathing-gas substitutions that have been reported to cause inner ear DCS. The model predicts considerable supersaturation, and therefore possible bubble formation, during the initial phase of a conventional decompression. Counterdiffusion of helium and nitrogen from the perilymph may produce supersaturation in the membranous labyrinth and endolymph after switching to a nitrogen-rich breathing mixture even without decompression. Conventional decompression algorithms may result in inadequate decompression for the inner ear for deep dives. Breathing-gas switches should be scheduled deep or shallow to avoid the period of maximum supersaturation resulting from decompression. PMID- 12562680 TI - Strength training reduces arterial blood pressure but not sympathetic neural activity in young normotensive subjects. AB - The effects of resistance training on arterial blood pressure and muscle sympathetic nerve activity (MSNA) at rest have not been established. Although endurance training is commonly recommended to lower arterial blood pressure, it is not known whether similar adaptations occur with resistance training. Therefore, we tested the hypothesis that whole body resistance training reduces arterial blood pressure at rest, with concomitant reductions in MSNA. Twelve young [21 +/- 0.3 (SE) yr] subjects underwent a program of whole body resistance training 3 days/wk for 8 wk. Resting arterial blood pressure (n = 12; automated sphygmomanometer) and MSNA (n = 8; peroneal nerve microneurography) were measured during a 5-min period of supine rest before and after exercise training. Thirteen additional young (21 +/- 0.8 yr) subjects served as controls. Resistance training significantly increased one-repetition maximum values in all trained muscle groups (P < 0.001), and it significantly decreased systolic (130 +/- 3 to 121 +/- 2 mmHg; P = 0.01), diastolic (69 +/- 3 to 61 +/- 2 mmHg; P = 0.04), and mean (89 +/- 2 to 81 +/- 2 mmHg; P = 0.01) arterial blood pressures at rest. Resistance training did not affect MSNA or heart rate. Arterial blood pressures and MSNA were unchanged, but heart rate increased after 8 wk of relative inactivity for subjects in the control group (61 +/- 2 to 67 +/- 3 beats/min; P = 0.01). These results indicate that whole body resistance exercise training might decrease the risk for development of cardiovascular disease by lowering arterial blood pressure but that reductions of pressure are not coupled to resistance exercise induced decreases of sympathetic tone. PMID- 12562681 TI - Sex differences in human skeletal muscle fatigue are eliminated under ischemic conditions. AB - Several studies have suggested that women may be more resistant to muscle fatigue than men (Fulco CS, Rock PB, Muza SA, Lammi E, Cymerman A, Butterfield G, Moore, LG, Braun B, and Lewis SF. Acta Physiol Scand 167: 233-239, 1999) possibly because of differences in muscle oxidative metabolism. We evaluated muscle fatigue produced by intermittent, maximal volitional isometric contractions of the dorsiflexor muscles of healthy young (21-34 yr) men (n = 8) and women (n = 8) under two conditions: free-flow (FF) circulation and ischemia. Measures of voluntary and stimulated (10- and 50-Hz) force, central activation ratio (CAR), and compound muscle action potential (CMAP) were collected in each session. The ischemic protocol induced greater fatigue than the FF protocol, in both sexes, and was associated with greater reductions in CAR, CMAP, stimulated force, and the ratio of 10- to 50-Hz force compared with the FF condition. Women fatigued less than men in FF but not during ischemia, and this difference was roughly paralleled by a difference in CAR. No sex effects on the CMAP, tetanic force, and measures of excitation-contraction coupling function were found in the FF condition, suggesting that the primary mechanism behind the difference in fatigue was a relatively greater impairment of central activation in men. The observation that ischemia eliminated the sex differences in fatigue is consistent with a number of studies (Kent-Braun JA, Ng AV, Doyle JW, and Towse TF. J Appl Physiol 93: 1813-1823, 2002) relating fatigue to muscle metabolism and might be the result of sex-based differences in metabolic pathway utilization during muscle contraction. PMID- 12562682 TI - HIV protease inhibitors: antiretroviral agents with anti-inflammatory, anti angiogenic and anti-tumour activity. PMID- 12562683 TI - Stopping HIV fusion with enfuvirtide: the first step to extracellular HAART. PMID- 12562684 TI - The use of tenofovir disoproxil fumarate for the treatment of nucleoside resistant HIV-1. PMID- 12562685 TI - Treatment of advanced HIV infection. PMID- 12562686 TI - Impact of clade diversity on HIV-1 virulence, antiretroviral drug sensitivity and drug resistance. AB - HIV-1 infection is characterized by genetic diversity wherein distinct viral subtypes (clades A, B, C, D, E, F, G, K and O) are expanding in different geographical regions. This article deals with the topic of HIV-1 subtype diversity in the context of sensitivity to antiretroviral drugs, drug resistance and viral fitness. Increasing evidence suggests that all clades of HIV probably display similar sensitivity to antiviral drugs. However, viruses from some subtypes and/or geographical regions may have a greater propensity to develop resistance against certain drugs than do other viral variants. In addition, differences in regard to replication capacity or fitness may exist among various HIV subtypes and differences in this regard may potentially become magnified under conditions of drug resistance. Immunological pressures may also play an important role in the evolution of viral subtypes that may impact on ultimate drug resistance profiles. PMID- 12562687 TI - Herbal medicines for treatment of bacterial infections: a review of controlled clinical trials. AB - OBJECTIVES: Many hundreds of plant extracts have been tested for in vitro antibacterial activity. This review is a critical evaluation of controlled clinical trials of herbal medicines with antibacterial activity. METHODS: Four electronic databases were searched for controlled clinical trials of antibacterial herbal medicines. Data were extracted and validated in a standardized fashion, according to predefined criteria, by two independent reviewers. RESULTS: Seven clinical trials met our inclusion criteria. Four of these studies were randomized. Three trials of garlic and cinnamon treatments for Helicobacter pylori infections reported no significant effect. Bacterial infections of skin were treated in four trials. Positive results were reported for an ointment containing tea leaf extract in impetigo contagiosa infections. Two trials of tea tree oil preparations used for acne and methicillin-resistant Staphylococcus aureus, and one trial of Ocimum gratissimum oil for acne, reported results equivalent to conventional treatments. CONCLUSIONS: Few controlled clinical trials have been published and most are methodologically weak. The clinical efficacy of none of the herbal medicines has so far been demonstrated beyond doubt. This area seems to merit further study through rigorous clinical trials. PMID- 12562688 TI - Physiological and molecular analysis of a mecA-negative Staphylococcus aureus clinical strain that expresses heterogeneous methicillin resistance. AB - Staphylococcus aureus clinical isolate 61/5896 exhibited methicillin resistance (MIC 64 mg/L), but lacked mecA, which encodes penicillin-binding protein 2'. The strain was isolated in England in 1961, and exhibited unstable heterogeneous methicillin resistance. When cultivated in drug-free medium, the methicillin resistance of 61/5896 increased after three daily passages, then decreased and was completely lost after 12 days' passage. Electron microscopy revealed that strain 61/5896 had a thicker and rougher cell wall than its methicillin susceptible derivatives. It produced about three times more penicillin-binding protein 2 (PBP2) than methicillin-susceptible derivatives. The strain was characteristically a non-producer of autolytic enzyme, though the phenotype, which was lost easily, was not directly correlated with methicillin resistance. PMID- 12562689 TI - On functional and structural heterogeneity of VIM-type metallo-beta-lactamases. AB - The VIM metallo-beta-lactamases are emerging resistance determinants, encoded by mobile genetic elements, that have recently been detected in multidrug-resistant nosocomial isolates of Pseudomonas aeruginosa and other Gram-negative pathogens. In this work a T7-based expression system for overproduction of the VIM-2 enzyme by Escherichia coli was developed, which yielded approximately 80 mg of protein per litre of culture. The enzyme was mostly released into the medium, from which it was recovered at >99% purity by an initial ammonium sulphate precipitation followed by two chromatography steps, with almost 80% efficiency. Determination of kinetic parameters of VIM-2 under the same experimental conditions previously used for VIM-1 (the first VIM-type enzyme detected in clinical isolates, which is 93% identical to VIM-2) revealed significant differences in K(m) values and/or turnover rates with several substrates, including penicillins, cephalosporins and carbapenems. Compared with VIM-1, VIM-2 is more susceptible to inactivation by chelators, indicating that the zinc ions of the latter are probably more loosely bound. These data indicated that at least some of the amino acid differences between the two proteins have functional significance. Molecular modelling of the two enzymes identified some amino acid substitutions, including those at positions 223, 224 and 228 (in the BBL numbering), that could be relevant to the changes in catalytic behaviour. PMID- 12562690 TI - Molecular and biochemical characterization of a carbapenem-hydrolysing beta lactamase from Flavobacterium johnsoniae. AB - Flavobacterium johnsoniae CIP100931 is resistant to most beta-lactam antibiotics and has a decreased susceptibility to carbapenems. A beta-lactamase gene was cloned and expressed in Escherichia coli DH10B. The purified beta-lactamase, JOHN 1, with a pI value of 9.0 and with a determined relative molecular mass of approximately 27 kDa was found to be a monomeric zinc-dependent enzyme that hydrolyses penicillins, narrow- and expanded-spectrum cephalosporins, carbapenems, but not monobactams. Sequence analysis revealed that JOHN-1 is a molecular class B beta-lactamase that is most closely related to BlaB from Chryseobacterium meningosepticum and IND-1 from Chryseobacterium indologenes (47% and 41% amino acid identity, respectively). JOHN-1 is a new member of the highly divergent subclass B1 lineage of metallo-enzymes. Although F. johnsoniae and Chryseobacterium spp. are phylogenetically related bacteria, this report further underlines the heterogeneity of class B beta-lactamases that are naturally produced by environmental Gram-negative aerobes and that are now recognized as the most important reservoir for these beta-lactamase genes. PMID- 12562691 TI - Development and mechanism of fluoroquinolone resistance in Legionella pneumophila. AB - The potential for selection in vitro of Legionella pneumophila mutants resistant to fluoroquinolones was investigated. Six distinct clinical isolates of L. pneumophila were subcultured in subinhibitory concentrations of ciprofloxacin, levofloxacin, clinafloxacin, trovafloxacin and moxifloxacin until MICs increased at least eight-fold. The numbers of serial passages required in microbroth dilution series were determined. The gyrA gene of the six parental strains, and 12 selected mutant strains, was sequenced. The five quinolones differed markedly in their ability to select mutants with decreased susceptibility. The average number of serial passages required was low in the cases of clinafloxacin (n = 10.6), ciprofloxacin and levofloxacin (both n = 13), but notably higher for trovafloxacin (n = 26.6) and moxifloxacin (n = 22.5). Five mutants treated with ciprofloxacin and three treated with moxifloxacin showed Thr83-->Lys or Thr83- >Ile amino acid changes in the gyrA gene. In conclusion, different quinolones lose their antimicrobial effect after a varying number of passages. This study demonstrated, for the first time to our knowledge, that gyrA in L. pneumophila is a possible target of fluoroquinolones. PMID- 12562692 TI - Treatment of plastic and extracellular matrix components with chlorhexidine or benzalkonium chloride: effect on Candida albicans adherence capacity in vitro. AB - This study investigates the influence of treatment of plastic and extracellular matrix (ECM) proteins with chlorhexidine or benzalkonium chloride on subsequent adherence of Candida albicans. Three concentrations were tested for each antiseptic: (i) chlorhexidine, MIC (6.25-12.5 mg/L), 80 x MIC and 800 x MIC; and (ii) benzalkonium chloride, MIC (3.12 mg/L), 40 x MIC and 1600 x MIC. Chlorhexidine and benzalkonium chloride activities were correlated with the tested concentrations. Antiseptics used at MIC were unable to modify the adherence to plastic or ECM proteins. Chlorhexidine (80 x MIC) induced a decrease in plastic adherence of 31% of the 15 strains used and an increase in ECM protein adherence of 13% of strains. Benzalkonium chloride (40 x MIC) induced a decrease in adherence to ECM proteins or plastic of 13-27% of strains. Our results indicated that the treatment with 1600 x MIC benzalkonium chloride could induce the opposite effect on adherence, depending on the surface: 60% of the strains showed an increase in their adherence to ECM proteins, whereas 93% of the strains showed a decrease in their adherence to plastic. A similar phenomenon was observed after treatment with 800 x MIC chlorhexidine: 60% of the strains showed an increase in their adherence to ECM proteins, whereas 67% showed a decrease in adherence to plastic. Treatment of medical devices with at least 5000 mg/L of chlorhexidine or benzalkonium chloride could therefore reduce C. albicans adherence to plastic surfaces, but would be unable to prevent fungal adherence to ECM proteins. PMID- 12562693 TI - Evaluation of selective and enrichment media for isolation of glycopeptide resistant enterococci from faecal specimens. AB - OBJECTIVES: Vancomycin-resistant enterococcus enrichment broth (VEB) and vancomycin-resistant enterococcus selective agar with vancomycin 6 mg/L (VSA) are novel azide-aesculin agar-based media that contain meropenem as an additional selective agent. The media were compared with enterococcosel broth (EB) and enterococcosel agar with vancomycin 6 mg/L (EA) for the isolation of glycopeptide resistant enterococci (GRE) from routine faecal screening specimens. METHODS: Two hundred and eighteen routine faecal screening specimens from patients at Addenbrooke's Hospital were examined. The majority were from patients on haematology wards (155) or the intensive therapy unit (ITU) (21). Specimens were inoculated on to VSA and EA directly, and after enrichment in VEB and EB, respectively. RESULTS: One hundred and twenty-eight GRE isolates were recovered from 93 (43%) specimens with enterococci carrying vanA or vanB genes. There were no statistically significant differences between media (specimens positive; numbers of GRE isolates) on direct plating on VSA (87; 104) or EA (86; 97) or following 24 h enrichment in VEB (89; 103) or EB (86; 98). There was no significant advantage to enrichment compared with direct plating. Incubation of enrichment broth cultures for only 6 h appeared detrimental. Enterococci with vanC were isolated significantly less frequently from VEB and VSA than from EB and EA. Growth of organisms other than GRE was more common on VSA than on EA. CONCLUSIONS: VEB and VSA were at least as effective as EB and EA for the recovery of GRE from faecal screening specimens, but substantially more non-GRE grew on VSA than on EA. Enrichment culture offered no significant advantages over direct plating. PMID- 12562694 TI - Comparison of three methods for in vitro susceptibility testing of Candida species with flucytosine. AB - Optimal methods for susceptibility testing of Candida spp. with flucytosine have not been determined. Breakpoints were recommended in 1984, but never validated. In this study, we compared the 1984 recommended macrodilution broth method (using an 80% endpoint) with a modification of the more recent NCCLS-recommended microdilution broth method with three endpoints-spectrophotometric 50% and 80% and a no growth endpoint determined by eye. NCCLS and British Society for Medical Mycology (BSMM) breakpoints were also compared. One hundred and fifty isolates comprised of Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, Candida parapsilosis and Candida lusitaniae were tested. Reproducibility was excellent. For C. albicans (n = 65), the correlation between tests was excellent (>75%), with few major discrepancies (<5%). For C. tropicalis (n = 27), correlation was good (59%), but there were a small number of major discrepancies (up to 11%, depending on breakpoint used). Results by the broth macrodilution method were generally higher than both microdilution methods for C. glabrata (n = 16; correlation of 18.8%), but only one major discrepancy was seen. Ten of the 11 C. parapsilosis isolates tested were susceptible by all methods, regardless of breakpoint chosen, with a correlation of 18.2%, but no major discrepancies were seen. A correlation between all methods (50%) was seen with C. lusitaniae (n = 10), with many isolates resistant or intermediate. In contrast, correlation between methods for C. krusei was poor (<5%); NCCLS microtitre modification produced results that were classified as intermediate or resistant, regardless of the breakpoint used. The methodology for susceptibility testing C. albicans is robust. Additional work to optimize susceptibility testing with flucytosine is necessary for non-albicans Candida species, especially C. krusei. PMID- 12562695 TI - The activity of amphotericin B against Candida albicans is not directly associated with extracellular calcium concentration. AB - The ability of amphotericin B to increase intracellular calcium concentrations in human cells is associated with the toxicity of this antifungal agent. The present study was performed to determine whether amphotericin B affects the influx or efflux of calcium in Candida albicans, and whether the antifungal activity of amphotericin B is dependent upon extracellular calcium concentrations. Concentration-response studies demonstrated that the addition of up to 1 mM EGTA to standard growth medium, with a more than 4000-fold decrease in extracellular calcium concentration, had no effect on the activity of amphotericin B against C. albicans. Amphotericin B did affect the kinetics of calcium influx acutely (< or =10 min), but had no net effect on long-term (1-24 h) calcium accumulation. Calcium efflux was also not affected by amphotericin B. These results indicate that, unlike its effects on mammalian cells, the toxicity of amphotericin B against C. albicans is not dependent upon increased movement of calcium across the cell membrane or the presence of extracellular calcium. PMID- 12562696 TI - Combination of caspofungin with inhibitors of the calcineurin pathway attenuates growth in vitro in Aspergillus species. AB - Work in Saccharomyces cerevisiae and Cryptococcus neoformans suggests that caspofungin could interact with the calcineurin pathway. We examined the in vitro interaction of caspofungin with calcineurin inhibitors (FK506, cyclosporin, FK520 and L685,818) and the TOR inhibitor rapamycin in 13 isolates of Aspergillus species. Caspofungin activity was enhanced by calcineurin/TOR inhibitors for all Aspergillus isolates studied. Further investigation of this pathway is warranted. PMID- 12562697 TI - Prevalence and types of class 1 integrons in aminoglycoside-resistant Enterobacteriaceae from several Chilean hospitals. AB - Members of the family Enterobacteriaceae are responsible for a variety of nosocomial infections, treatment of which is limited due to their increasing resistance to antibiotics. Some bacterial genes encoding antibiotic resistance comprise the major part of gene cassettes, most of which are associated with integrons. In this work, the carriage of class 1, 2 and 3 integrons was investigated in 191 Enterobacteriaceae isolates from clinical specimens of hospitalized patients. Class 1 integrons were found to be the most common, whereas no class 3 integrons were detected. The variable regions of 13 class 1 integrons were characterized and four types were found. Type 1 harbours only ant(3")I, type 2 harbours ant(2")I and ant(3")I, type 3 harbours aac(6')Ib and ant(3")I and type 4 lacks inserted gene cassettes. PMID- 12562698 TI - Prevalence of resistance to ampicillin, gentamicin and vancomycin in Enterococcus faecalis and Enterococcus faecium isolates from clinical specimens and use of antimicrobials in five Nordic hospitals. AB - We determined the species distribution and prevalence of ampicillin resistance, high-level gentamicin resistance (HLGR) and vancomycin resistance among clinical enterococcal isolates from five Nordic laboratories (Bergen, Tromso, Uppsala, Aarhus and Reykjavik). Isolates represented three different groups: (i) all blood culture isolates from 1999; (ii) consecutive in-patient isolates (maximum 40); and (iii) consecutive outpatient isolates (maximum 40) collected during March to May 2000. Antimicrobial use data were collected at the national and hospital level. A high proportion (31.4%) of Enterococcus faecium was detected among blood culture isolates, in contrast to only 4.2% among isolates from outpatients. Ampicillin resistance was not found in Enterococcus faecalis, in contrast to 48.8% in E. faecium isolates. HLGR rates varied considerably between laboratories (1.1-27.6%). Acquired vancomycin resistance was not detected. There were no significant differences in the prevalences of HLGR between in-patient and outpatient isolates at individual hospitals. A cluster of clonally related ampicillin-resistant and HLGR E. faecium isolates was demonstrated in one of the hospitals. The lowest level of hospital antimicrobial use, the lowest proportion of E. faecium and the lowest prevalence of resistance were observed in Reykjavik. The study showed a relatively low level of resistance in enterococci, as compared with most European countries and the USA. However, there were large differences between hospitals with regard to the relative proportion of E. faecium isolates, their susceptibility to ampicillin and gentamicin, as well as the prevalence of HLGR in E. faecalis isolates. This indicates a potential for further improvement of antibiotic policies, and possibly hospital infection control, to maintain the low resistance levels observed in these countries. PMID- 12562699 TI - Significant increase in the prevalence of erythromycin-resistant, clindamycin- and miocamycin-susceptible (M phenotype) Streptococcus pyogenes in Spain. AB - In 1998 we conducted a multicentre study in Spain on the susceptibility of Streptococcus pyogenes isolates to different 14-, 15- and 16-membered macrolides and clindamycin, in which the number of strains examined was proportional to the number of inhabitants in each geographical area. The aim of the present work was to re-examine the antimicrobial susceptibility of S. pyogenes in 2001, using the same methodology and centres as in 1998, to determine the different susceptibility phenotypes to macrolides-lincosamides, and to compare the results from the 2 years by statistical tests. A total of 529 unique isolates of S. pyogenes, collected in 21 laboratories, were studied. Throat swabs provided 417 isolates (78.8%), and the remaining 112 were from other sources. Four hundred and thirty-five (82.2%) were isolated from children and 94 (17.8%) from adults. One hundred and fifty-seven (29.7%) of the isolates were resistant to erythromycin and azithromycin, whereas resistance to miocamycin, a 16-membered macrolide, was 1.5%. The prevalence of resistance to clindamycin was 1.3%. The majority (98.7%) of the 157 erythromycin-resistant strains presented the M phenotype. When we compared the results obtained in 1998 and 2001, we observed a statistically significant increase in resistance to erythromycin and azithromycin (P = 0.02, chi(2) test), but not to clindamycin or miocamycin (P = 0.47, chi(2) test with Yates' correction). The significant increase in the prevalence of resistance to some macrolides of S. pyogenes in Spain underscores the need for continuous surveillance of antimicrobial resistance in this species. PMID- 12562700 TI - Multicentre evaluation of the in vitro activity of linezolid in the Western Pacific. AB - Multiresistance to antimicrobial agents is common in staphylococci and pneumococci isolates in the Western Pacific region. The activity of linezolid, a new oxazolidinone, was evaluated against a spectrum of Gram-positive species collected in the region. Eighteen laboratories from six countries in the Western Pacific examined the linezolid susceptibility of 2143 clinical isolates of Staphylococcus aureus, coagulase-negative staphylococci (CoNS) and Enterococcus spp. using broth microdilution or disc diffusion methodology. For Streptococcus pneumoniae (n = 351) and other streptococci (n = 83), Etest (AB Biodisk, Solna, Sweden) strips were used. Results were compared with other common and important antimicrobials. Linezolid-resistant strains were not detected among streptococci or staphylococci, including a significant proportion of S. aureus strains that were multiresistant. Almost all enterococci, including 14 vancomycin-resistant Enterococcus faecium, were linezolid susceptible. A small proportion of enterococci (0.8%) were intermediate to linezolid, and one strain of Enterococcus faecalis had a zone diameter of 20 mm (resistant). The linezolid MIC ranges (MIC(90)) of those strains tested by broth microdilution or Etest were: 1-4 mg/L (2 mg/L) for S. aureus, 0.5-4 mg/L (2 mg/L) for CoNS, 0.5-4 mg/L (2 mg/L) for Enterococcus spp., 0.12-2 mg/L (1 mg/L) for S. pneumoniae and 0.25-2 mg/L (1 mg/L) for Streptococcus spp. There was no difference in linezolid susceptibility between countries or between multiresistant and susceptible strains of each species monitored. PMID- 12562701 TI - Multicentre surveillance of Pseudomonas aeruginosa susceptibility patterns in nosocomial infections. AB - OBJECTIVES: To determine susceptibility rates and patterns in Pseudomonas aeruginosa strains isolated from nosocomial infections. METHODS: Seven hundred and sixteen P. aeruginosa isolates from 40 different hospitals in Belgium and the Grand Duchy of Luxembourg were collected in 1999. RESULTS: Resistance rates varied significantly between hospitals. Of the fluoroquinolones, ciprofloxacin showed least resistance (24%), levofloxacin showed 27.5% resistance and ofloxacin 37.5%. Of the aminoglycosides, amikacin was the most potent antibiotic (10.5% resistance), followed by isepamicin (12%), tobramycin (19.5%) and gentamicin (23.5%). Of the beta-lactam antibiotics, meropenem was the most active (9.5% resistance); piperacillin and piperacillin/tazobactam had, respectively, 24% and 17.5% resistance, ceftazidime 28.5%, cefepime 29.5%, ticarcillin/clavulanic acid 37% and aztreonam 55.5%. MIC distribution curves show the presence of significant subpopulations, with MICs just below breakpoint for many antibiotics. CONCLUSION: Resistance of P. aeruginosa to penicillins, cephalosporins, fluoroquinolones and aminoglycosides varies between hospitals, but is increasing. PMID- 12562702 TI - Antimicrobial-induced release of endotoxin from Pseudomonas aeruginosa: comparison of in vitro and animal models. AB - This study was designed to compare the amount of lipopolysaccharide (LPS) induced following exposure to doripenem, imipenem/cilastatin, meropenem and ceftazidime in an in vitro computerized-simulation system (simulating the drug concentration pattern in human plasma after administration of a drug), with that induced by exposure to a drug at a constant concentration. When Pseudomonas aeruginosa was exposed to the test drugs at constant concentrations of 0.1 x, 1 x and 10 x MIC, differential relative induction of LPS was observed as follows: ceftazidime > meropenem, doripenem > imipenem/cilastatin. In the computerized-simulation system, however, the amount of LPS induced by treatment with ceftazidime (1 g) was similar to that by doripenem (250 mg), imipenem/cilastatin (500 mg) and meropenem (500 mg). In a rat model of P. aeruginosa bacteraemia, rates of eradication of bacteria from the blood were similar for carbapenems and ceftazidime except for 1 h post-administration of ceftazidime. Serum LPS levels induced by treatment with doripenem (30 mg/kg), imipenem/cilastatin (30 mg/kg), meropenem/cilastatin (30 mg/kg) and ceftazidime (50 mg/kg) were almost the same at 3 h after administration of each drug. Data obtained from computerized simulation systems might be more applicable than those obtained from organisms exposed to constant drug concentrations for estimating the amount of LPS in the plasma of human patients infected with Gram-negative bacteria. PMID- 12562703 TI - Enzymic degradation of a beta-lactam antibiotic, ampicillin, in the gut: a novel treatment modality. AB - Antibiotics can cause severe alterations in the gut microflora and promote diarrhoea and overgrowth of pathogenic bacteria. The present study investigated the potency of targeted recombinant beta-lactamase (TRBL) to degrade a beta lactam antibiotic in the jejunum of fistula-operated beagles. We used different peroral doses of purified beta-lactamase (PenP) of Bacillus licheniformis in enteric-coated pellets together with intravenous ampicillin. Serum and jejunal samples were collected for ampicillin and beta-lactamase analysis. A dose response effect of TRBL on ampicillin concentrations in the jejunal samples could be observed. The highest doses applied decreased the jejunal ampicillin concentrations to undetectable levels. In the serum samples, the ampicillin concentrations were not affected by the beta-lactamase dose used. Our results indicate that it may be possible to evolve a targeted treatment to degrade beta lactam antibiotics intestinally and, thus, decrease antibiotic-induced adverse effects on the gut microflora. PMID- 12562704 TI - Monotherapy with mastic does not eradicate Helicobacter pylori infection from mice. AB - OBJECTIVE: To determine the ability of mastic monotherapy to eradicate Helicobacter pylori infection from mice. MATERIALS AND METHODS: The susceptibility of H. pylori SS1 to mastic was assessed by broth dilution determination of the MIC and MBC. Mice were inoculated intragastrically with either a suspension of H. pylori SS1 (n = 70) or brain-heart infusion broth alone (n = 10). Mice were given antimicrobial chemotherapy 4 weeks after infection and were administered the mouse equivalent of either 2 g of mastic twice daily for 7 days or a triple therapy regimen containing the mouse equivalent of 400 mg of metronidazole, 250 mg of clarithromycin and 20 mg of omeprazole twice daily for 7 days. Mice were killed either immediately or 1 month after the completion of treatment, and their stomachs cultured for H. pylori. RESULTS: The mastic MIC and MBC of H. pylori SS1 were 7.80 and 31.25 mg/L, respectively. The triple therapy regimen eradicated infection from 19 of 20 SS1-infected mice. Mastic failed to eradicate infection from any of the 18 SS1-infected mice (P < 0.001) and there was no signifi- cant reduction in gastric bacterial load in mice treated with this regimen. CONCLUSION: Despite reported beneficial effects in ulcer patients and the good in vitro activity of mastic against H. pylori, this compound is unable to eradicate H. pylori infection from mice. PMID- 12562705 TI - Identical pattern of highly variable absorption of clavulanic acid from four different oral formulations of co-amoxiclav in healthy subjects. AB - The aims of this investigation were to calculate the pharmacokinetic parameters of amoxicillin and clavulanic acid, and to identify parameters that may affect their observed differences in absorption. Data were obtained from plasma concentration-time curves from four different open, randomized, two-treatment, two-period, two-sequence, crossover Phase I bioequivalence studies, with the following co-amoxiclav formulations: tablets 250/125, 500/125 and 875/125 mg, or 10 mL of an oral suspension 250/62.5 mg per 5 mL. Data from 144 subjects and 288 drug administrations were available for evaluation. After a 125 mg clavulanic acid dose (administered as potassium clavulanate) for all four different formulations, the clavulanic acid AUC(t) data ranged from 1.5 to 8 mg.h/L, varying by a factor of 5. The absorption of clavulanic acid was not related to the absorption of amoxicillin, or demographic factors, and we were unable to identify the reasons for the large variability in the absorption of clavulanic acid. We conclude that the absorption of clavulanic acid, after oral administration, is highly variable and may vary over a five-fold range between patients. PMID- 12562706 TI - Laboratory antibiotic susceptibility reporting and antibiotic prescribing in general practice. AB - This study set out to investigate whether there was an association between antibiotic susceptibility reporting from microbiology laboratories and antibiotic prescribing for urinary tract infections in the community. Data were collected over a 3 month period using a prospective questionnaire survey of general practitioners, who submitted and received a mid-stream urine (MSU) result from selected microbiology laboratories in England and Wales. In addition, prescribing analyses and cost (PACT) data were requested from the Prescription Prescribing Authority. The study demonstrated an association between laboratory reporting of antibiotic susceptibilities and antibiotic prescribing for treatment of urinary tract infections. The reporting of susceptibilities to oral cephalosporins and nitrofurantoin from microbiology laboratories was associated with increased prescribing of each antibiotic. This association was demonstrated for the choice of empirical antibiotic therapy and the choice of antibiotic prescribed for each studied episode of urinary tract infection. PACT data demonstrated a consistently greater use of antibiotics that were reported by the servicing laboratory, although this was only statistically significant for nitrofurantoin. This study demonstrates that there is an association between antibiotic susceptibility reporting from microbiology laboratories and antibiotic prescribing for the treatment of urinary tract infections. PMID- 12562707 TI - Influence of population density on antibiotic resistance. AB - Antibiotic consumption and population density as a measure of crowding in the community were related to the prevalence of antibiotic resistance of three cities in three different countries: St Johns in Newfoundland (Canada), Athens in Greece and Groningen in The Netherlands. Antibiotic consumption was expressed in DDD (defined daily dose), as DID (DDD/1000 inhabitants/day) and as DSD (DDD/km(2)). The prevalence of antibiotic-resistant Escherichia coli and enterococci was determined in faecal samples of healthy volunteers. In both Newfoundland (28 DID) and Greece (29 DID) the overall consumption of antibiotics was more than three times higher compared with that of The Netherlands (9 DID). The lowest prevalence of resistant E. coli against the majority of antibiotics tested was found for the samples from Newfoundland and was significant (P < 0.05) for cefazolin, oxytetracycline and trimethoprim. A poor correlation between the number of DID and the prevalence of resistance was observed [the Pearson correlation coefficient (Pcc) ranged between -0.93 and 0.87]. However, when population density was taken into consideration and antibiotic consumption was expressed in DSD, a strong correlation was observed (and Pcc ranged between 0.86 and 1.00). This study suggests that population density is an important factor in the development of antibiotic resistance and warrants special attention as a factor in resistance epidemiology. PMID- 12562708 TI - Cost-minimization analysis and audit of antibiotic management of bone and joint infections with ambulatory teicoplanin, in-patient care or outpatient oral linezolid therapy. AB - Bone and joint infections are significant causes of morbidity, mortality and healthcare costs. The cost of treatment for such infections is driven primarily by the length of hospital stay. Many of these infections will require treatment with prolonged periods of parenteral antibiotic therapy. Clinicians and healthcare managers are being attracted increasingly by administering treatment in the ambulatory setting as this offers clinical, economic and quality of life advantages from both the hospital's and patient's perspective. Our retrospective audit of managing 55 treatment episodes of bone and joint infections with teicoplanin delivered in the outpatient or home setting revealed that the mean cost of care per episode of infection was less with treatment in the ambulatory setting ( pound 1749.15) compared with the in-patient setting ( pound 11 400) or compared with the hypothetical situation of treatment with oral linezolid in the home setting ( pound 2546). Teicoplanin therapeutic drug monitoring appears to be valuable in establishing optimal serum levels, which appear to correlate with good clinical outcomes. The potential for alternative day or thrice weekly dosing with teicoplanin may offer further cost advantages whilst maintaining equivalent clinical effectiveness. PMID- 12562709 TI - Characterization of a new integron containing VIM-2, a metallo- beta-lactamase gene cassette, in a clinical isolate of Enterobacter cloacae. AB - We report the first description of a new integron containing bla(VIM-2), a metallo-beta-lactamase gene from Enterobacter cloacae KU680, which was isolated from peritoneal fluid of a liver cirrhosis patient in South Korea. Antibiotic susceptibility testing, and modified Hodge and EDTA-disc synergy tests, were carried out to screen for metallo-beta-lactamase-producing strains. PCR and sequence analysis were used to identify and analyse the bla(VIM-2)-containing integron. The isolate was resistant to most beta-lactams, including imipenem, and demonstrated a positive modified Hodge and EDTA-disc synergy test, which are findings suggesting a metallo-beta-lactamase. Preliminary PCR-based experiments detected the metallo-beta-lactamase gene bla(VIM-2). Sequencing of the 4392 bp cloned PCR amplicon, containing the gene cassette bla(VIM-2), revealed the structure of the class 1 integron. The integron also contained additional insert gene cassettes, aadA, and unknown open reading frames 'orfII' and 'orfIII'. To the best of our knowledge, this is the first time that this metallo-beta lactamase gene has been detected in E. cloacae. PMID- 12562710 TI - Effect of ciprofloxacin on adhesive properties of non-P mannose-resistant uropathogenic Escherichia coli isolates. AB - The influence of sub-MIC of ciprofloxacin on the surface properties of 25 non-P mannose-resistant uropathogenic Escherichia coli (UPEC) strains was studied. Thirteen isolates responded to antibiotic treatment with an increase in haemagglutination titre and/or surface hydrophobicity, which correlated with a higher expression of surface proteins. Only UPEC strains with ciprofloxacin enhanced hydrophobicity increased their adhesiveness to urinary catheters that correlated, in one analysed case, with a dramatic increase in the number of fimbriae peripherally located. The overall results indicate that sub-MICs of ciprofloxacin could increase the adhesiveness, and hence the risk of colonization by UPEC strains expressing mannose-resistant adhesins different from type P. PMID- 12562711 TI - Heat-induced superaggregation of amphotericin B attenuates its ability to induce cytokine and chemokine production in the human monocytic cell line THP-1. AB - The cytokine and chemokine response elicited by heat-treated amphotericin B (HT AmB) was compared with that of untreated amphotericin B (AmB-DOC) in the human monocyte cell line THP-1. AmB-DOC produced dose-dependent increases in interleukin (IL)-1beta, IL-1alpha, tumour necrosis factor-alpha, macrophage inflammatory protein (MIP)-1alpha and MIP-1beta at 2 h. HT-AmB induced cytokine and chemokine production at a lower level than those observed with corresponding concentrations of AmB-DOC, while retaining antifungal activity. These results indicate that heat treatment of amphotericin B may prove to be a cost-effective approach to improving the therapeutic index of this antifungal agent. PMID- 12562712 TI - Two major Spanish clones of penicillin-resistant Streptococcus pneumoniae in Portuguese isolates of clinical origin. AB - We studied the genetic relatedness of 47 Portuguese penicillin-resistant 9V and 23F Streptococcus pneumoniae of clinical origin, using pulsed-field gel electrophoresis (PFGE) and restriction fragment length polymorphism analysis. PFGE fingerprinting showed that 24 isolates of serotype 9V and 23 isolates of serotype 23F were variants of the Spain(9V)-3 and Spain(23F)-1 clones, respectively. Fingerprinting of pbp1a, pbp2b and pbp2x genes showed that all the penicillin-resistant clones gave similar HinfI restriction patterns. In this study, serotypes 9V and 23F have different clonal origins and identical PBP genotypes, suggesting a horizontal transfer of resistance. Visual and computer assisted analysis of PFGE patterns correlated well (r = 0.983). PMID- 12562713 TI - Occurrence and detection of AmpC beta-lactamases among Gram-negative clinical isolates using a modified three-dimensional test at Guru Tegh Bahadur Hospital, Delhi, India. AB - AmpC enzymes can be differentiated from other extended-spectrum beta-lactamases by their ability to hydrolyse cephamycins as well as other extended-spectrum cephalosporins. The present study was designed to determine the occurrence of AmpC enzyme-harbouring Gram-negative clinical isolates in a tertiary care hospital in Delhi, India. Among the 135 clinical isolates of Gram-negative bacilli tested, 20.7% were found to harbour AmpC enzymes using a modified three dimensional test. Inhibition of zone distortion in the presence of cloxacillin confirmed AmpC-harbouring isolates. Maximal incidence of AmpC producers was found among Acinetobacter spp. (42.8%) followed by Klebsiella pneumoniae isolates (33.3%). No AmpC-harbouring isolates revealed decreased susceptibility to cefoxitin. Therefore, Gram-negative bacilli showing resistance to any cephalosporin or aztreonam irrespective of cefoxitin susceptibility should be screened for the AmpC enzyme. The modified three-dimensional test is easy to carry out and can be applied as a phenotypic screening method for detection of AmpC-harbouring Gram-negative organisms. This is the first study to determine the occurrence of AmpC beta-lactamases from India. PMID- 12562714 TI - Detection of methicillin resistance in coagulase-negative staphylococci and in staphylococci directly from simulated blood cultures using the EVIGENE MRSA Detection Kit. AB - The EVIGENE MRSA Detection Kit was evaluated on coagulase-negative staphylococci (CoNS) from agar plates and on staphylococci directly from positive spiked blood cultures. For the CoNS study, a total of 242 isolates were tested, and of these 237 gave valid test results. For the 237 valid tests, all gave correct mecA classification. For the blood culture procedure, a collection of 51 mecA-positive Staphylococcus aureus, 21 mecA-negative S. aureus, 31 mecA-positive CoNS and 28 mecA-negative CoNS were used for the simulated blood cultures. For the S. aureus strains, all gave valid test results and correct mecA classification. One of the MRSA isolates gave a very faint nuc signal, and another four isolates gave results close to the cut-off of the kit; however, these were still clearly positive when read by the naked eye. For the CoNS isolates, 51 of the 59 strains gave valid results. All of these 51 strains gave correct mecA status. Thus the EVIGENE MRSA Detection Kit can provide fast and accurate determination of methicillin resistance in CoNS. This preliminary study of the blood culture procedure indicates that it is possible to achieve determination of methicillin resistance in staphylococci 8 h after positivity of the blood culture, making same-day detection of methicillin resistance possible. PMID- 12562715 TI - Ex vivo synergy of arachidonate-enriched serum with ceftazidime and amikacin on multidrug-resistant Pseudomonas aeruginosa. AB - Three multidrug-resistant strains of Pseudomonas aeruginosa were incubated ex vivo with sera sampled after a 10 min intravenous infusion of 25 mg/kg of arachidonic acid (AA) in 10 rabbits in the presence of ceftazidime and amikacin. Lipid peroxidation was assessed during bacterial growth. A statistically significant decrease in bacterial cells was found by the interaction of antimicrobials and serum sampled in the middle of infusion and 15 and 30 min after infusion of AA and was accompanied by elevated levels of malonodialdehyde. This effect of AA is probably attributed to lipid peroxidation and raises the possibility of its application in experimental infections. PMID- 12562716 TI - Growing antimicrobial resistance of Shigella isolates. AB - Appropriate antibiotic treatment of shigellosis depends on identifying resistance patterns. By studying Shigella isolates in central Israel from 1998 to 2000, and comparing with the period 1991-1992, we identified a significantly increased resistance to tetracycline (from 23% to 87%, P < 0.00001), high resistance to trimethoprim-sulfamethoxazole (94%) and ampicillin (85%) and emerging resistance to quinolones (0.5-2%). These data mandate local monitoring of resistance and its consideration in empirical therapy. PMID- 12562717 TI - Antifungal susceptibility patterns of yeast isolates causing bloodstream infections. AB - A broth microdilution method following the NCCLS recommendations was used for testing fluconazole, itraconazole, posaconazole, 5-fluorocytosine and amphotericin B against 83 yeast isolates causing bloodstream infections in 59 patients hospitalized between January 1998 and June 2001 at the University Hospital of Bari, Italy. Isolates belonged to four species of Candida and three other yeast genera. Of the isolates, 97%, 95% and 100% were susceptible to fluconazole, itraconazole and posaconazole, respectively. Similarly, 97% and 100% of the isolates were susceptible to 5-fluorocytosine and amphotericin B, respectively. PMID- 12562718 TI - Antimycobacterial activity of diospyrin derivatives and a structural analogue of diospyrin against Mycobacterium tuberculosis in vitro. AB - Three derivatives and one structural analogue of diospyrin were synthesized and investigated for their inhibitory activity against Mycobacterium tuberculosis employing the rapid radiometric method in vitro. A novel aminoacetate derivative was found to be more active than the parent compound, the MICs being 50 and 100 mg/L, respectively, for a drug-susceptible strain, H37Rv, of M. tuberculosis. This derivative also exhibited an MIC of 50 mg/L for a few multidrug-resistant strains of M. tuberculosis. The other two derivatives and the analogue did not show any significant antimycobacterial activity at the highest concentration (100 mg/L) tested. PMID- 12562719 TI - Antimicrobial activity of esomeprazole versus omeprazole against Helicobacter pylori. AB - OBJECTIVE: Esomeprazole is an enantiomorph of omeprazole, which inhibits gastric acid secretion more effectively than omeprazole. As proton pump inhibitors also exert an antibacterial activity, we aimed to compare esomeprazole and omeprazole for their antimicrobial activity against Helicobacter pylori in vitro. METHODS: We studied 52 H. pylori isolates obtained from gastric biopsies and inoculated onto agar plates containing the acid-converted drugs at different concentrations. The minimal concentrations that inhibited the growth of 50% and 90% of isolates were defined as MIC(50) and MIC(90). RESULTS: The MIC(50) and MIC(90) of esomeprazole were 16 and 32 mg/L; and those of omeprazole were 32 and 64 mg/L. Overall, 63.5% of isolates showed the same susceptibility to both drugs; 17 isolates were two- to 64-fold more susceptible to esomeprazole and two isolates were two-fold more susceptible to omeprazole. CONCLUSIONS: The increased antimicrobial activity in vitro of esomeprazole against H. pylori could contribute to improving the outcome of the eradication treatment of such an infection. PMID- 12562720 TI - Bactericidal activity of daptomycin against Streptococcus pneumoniae compared with eight other antimicrobials. AB - A spectrum of pneumococci with varying susceptibilities to beta-lactams, macrolides and quinolones was tested for susceptibility to nine antibiotics, including the novel lipopeptide daptomycin. Daptomycin was active against all strains (MIC range 90% killing at 1 h. PMID- 12562721 TI - A national surveillance of antimicrobial resistance in Escherichia coli isolated from food-producing animals in Japan. AB - A nationwide investigation of antimicrobial susceptibility in Escherichia coli isolated from food-producing animals was performed in Japan. MICs of 18 antimicrobial agents were determined for a total of 1018 isolates. Higher resistance rates were observed against sulfadimethoxine, oxytetracycline and dihydrostreptomycin, followed by ampicillin and kanamycin. Resistance was more frequently observed among broiler isolates, followed by isolates from pigs. Almost 10% of broiler isolates were resistant to fluoroquinolones and extremely high MICs (100 mg/L) were observed. In general, antimicrobial resistance rates in E. coli have declined in recent years, with the exception of resistance to fluoroquinolones among broiler isolates, which has increased. PMID- 12562722 TI - A randomized comparative study to determine the effect of omeprazole on the peak serum concentration of itraconazole oral solution. AB - To determine the effect of omeprazole on peak serum concentrations (C(max)) of itraconazole oral solution (IOS), we carried out a randomized, open-label, prospective, crossover study. Fifteen healthy, non-pregnant adults received a single dose of IOS 400 mg on two occasions, at least 7 days apart, with omeprazole 40 mg nightly for 7 days before either IOS dose 1 or 2. C(max), time to C(max) (T(max)) and AUC(0-8) were determined for itraconazole and its active metabolite, hydroxyitraconazole, for each dose and compared. Omeprazole did not significantly affect the C(max), T(max) or AUC(0-8) of itraconazole or hydroxyitraconazole when administered as IOS. PMID- 12562723 TI - Tissue and serum concentrations of levofloxacin 500 mg administered intravenously or orally for antibiotic prophylaxis in biliary surgery. AB - OBJECTIVES: Levofloxacin is a third-generation fluoroquinolone with a broad spectrum of antibacterial activity, comprising enterobacteria, non-fermenters, Gram-positive cocci and some anaerobic species. Members of these species are common pathogens in acute and chronic cholecystitis. This suggests that levofloxacin may be used as peri-operative prophylaxis in gall-bladder surgery. The purpose of our study was to determine serum and tissue levels of levofloxacin in cholecystectomy patients following pre-operative dosing. PATIENTS AND METHODS: Patients with gall-bladder surgery were given levofloxacin 500 mg as a single dose either intravenously (iv) or orally pre-operatively, at the treating physician's decision. Gall-bladder tissue and serum samples were collected, and drug concentrations were determined by HPLC with fluorescence detection. Additionally, all tissue samples underwent routine microbiological diagnostics. MICs for aerobic isolates were determined using the Etest. RESULTS: A total of 61 patients (48 female, 13 male) were included. The medians of the levofloxacin concentrations in serum were 11.37 mg/L (iv) and 9.65 mg/L (oral), and in gall bladder tissue they were 15.61 mg/kg (iv) and 17.93 mg/kg (oral). Eleven pathogens were isolated from gall-bladder samples. Post-operative wound infection was observed in two of the 61 patients. CONCLUSION: Our data suggest that levofloxacin may be considered for peri-operative prophylaxis in biliary tract surgery. PMID- 12562724 TI - Dual activity of fluoroquinolones against Streptococcus pneumoniae. PMID- 12562726 TI - Variation in the content of ciprofloxacin susceptibility testing discs: its effect on the BSAC disc method. PMID- 12562727 TI - Importance of using current NCCLS breakpoints to interpret cefotaxime and ceftriaxone MICs for Streptococcus pneumoniae. PMID- 12562728 TI - Detection of intrinsic oxacillin resistance in non-multiresistant, oxacillin resistant Staphylococcus aureus (NORSA). PMID- 12562729 TI - CTX-M extended-spectrum beta-lactamase arrives in the UK. PMID- 12562730 TI - First isolation of CTX-M15-producing Escherichia coli from two French patients. PMID- 12562731 TI - Persistent acute tubular toxicity after switch from conventional amphotericin B to liposomal amphotericin B (Ambisome). PMID- 12562732 TI - The Human Rights Act and mental health legislation. PMID- 12562733 TI - Psychiatric genetics in the post-genome age. PMID- 12562734 TI - Neurobiology of addiction and implications for treatment. PMID- 12562735 TI - The relativity of relative risks: disadvantage or opportunity? PMID- 12562736 TI - For: does workplace counselling work? PMID- 12562737 TI - Ethnic variations in pathways to and use of specialist mental health services in the UK. Systematic review. AB - BACKGROUND: Inequalities of service use across ethnic groups are important to policy makers, service providers and service users. AIMS: To identify ethnic variations in pathways to specialist mental health care, continuity of contact, voluntary and compulsory psychiatric in-patient admissions; to assess the methodological strength of the findings. METHOD: A systematic review of all quantitative studies comparing use of mental health services by more than one ethnic group in the UK. Narrative analysis supplemented by meta-analysis, where appropriate. RESULTS: Most studies compared Black and White patients, finding higher rates of in-patient admission among Black patients. The pooled odds ratio for compulsory admission, Black patients compared with White patients, was 4.31 (95% CI 3.33-5.58). Black patients had more complex pathways to specialist care, with some evidence of ethnic variations in primary care assessments. CONCLUSIONS: There is strong evidence of variation between ethnic groups for voluntary and compulsory admissions, and some evidence of variation in pathways to specialist care. PMID- 12562738 TI - Migration as a risk factor for schizophrenia: a Danish population-based cohort study. AB - BACKGROUND: A growing body of evidence suggests that migration is a risk factor for the development of schizophrenia, although the putative mechanism remains obscure. AIMS: To examine immigrant background and history of foreign residence as risk factors for schizophrenia. METHOD: Using data from the Danish Civil Registration System, we established a population-based cohort of 2.14 million persons resident in Denmark by their fifteenth birthday. Schizophrenia in cohort members and parental psychiatric disorder were identified by cross-linkage with the Danish Psychiatric Case Register. RESULTS: The relative risk of developing schizophrenia was 2.45 (95% Cl 2.25-2.67) and 1.92 (95% Cl 1.74-2.12) among first and second-generation immigrants respectively, and 1.60 (95% Cl 1.25-2.05) among Danes with a history of foreign residence. CONCLUSIONS: Migration confers an increased risk for schizophrenia that is not solely attributable to selection factors and may also be independent of foreign birth. PMID- 12562739 TI - Predicting engagement with services for psychosis: insight, symptoms and recovery style. AB - BACKGROUND: Treatment non-adherence and service disengagement are commonly attributed to impaired insight. There is evidence that recovery style (i.e. psychological adjustment) may underlie service engagement. AIMS: We examined whether insight, psychotic symptoms or individuals' recovery style ('integration' v. 'sealing-over') predicts service engagement. METHOD: Fifty patients with schizophrenia were assessed during acute psychosis and at 3-month and 6-month follow-ups. Measures included recovery style, psychosis symptoms, insight and service engagement. RESULTS: Sealing-over at 3 months following onset of an episode of psychosis predicted low service engagement at 6 months. Neither insight nor symptoms predicted engagement. The clear shift from integration to sealing-over within the first 3 months was independent of changes in symptoms or insight. Sealing-over between 3 and 6 months was associated with improvement in psychosis symptoms. CONCLUSIONS: Recovery style contributed more to engagement than did insight, appears to be dynamic in the short term and is orthogonal to insight. Overall, this study demonstrated the importance of addressing psychological adjustment to psychosis as well as illness status when investigating treatment engagement in people with psychosis. PMID- 12562740 TI - Impact of comorbid personality disorder on violence in psychosis: report from the UK700 trial. AB - BACKGROUND: The impact of comorbid personality disorder on the occurrence of violence in psychosis has not been fully explored. AIMS: To examine the association between comorbid personality disorder and violence in community dwelling patients with psychosis. METHOD: A total of 670 patients with established psychotic illness were screened for comorbid personality disorder. Physical assault was measured from multiple data sources over the subsequent 2 years. Logistic regression was used to assess whether the presence of comorbid personality disorder predicted violence in the sample. RESULTS: A total of 186 patients (28%) were rated as having a comorbid personality disorder. Patients with comorbid personality disorder were significantly more likely to behave violently over the 2-year period of the trial (adjusted odds ratio=1.71, 95% CI 1.05-2.79). CONCLUSIONS: Comorbid personality disorder is independently associated with an increased risk of violent behaviour in psychosis. PMID- 12562741 TI - Dialectical behaviour therapy for women with borderline personality disorder: 12 month, randomised clinical trial in The Netherlands. AB - BACKGROUND: Dialectical behaviour therapy (DBT) is widely considered to be a promising treatment for borderline personality disorder (BPD). However, the evidence for its efficacy published thus far should be regarded as preliminary. AIMS: To compare the effectiveness of DBT with treatment as usual for patients with BPD and to examine the impact of baseline severity on effectiveness. METHOD: Fifty-eight women with BPD were randomly assigned to either 12 months of DBT or usual treatment in a randomised controlled study. Participants were recruited through clinical referrals from both addiction treatment and psychiatric services. Outcome measures included treatment retention and the course of suicidal, self-mutilating and self-damaging impulsive behaviours. RESULTS: Dialectical behaviour therapy resulted in better retention rates and greater reductions of self-mutilating and self-damaging impulsive behaviours compared with usual treatment, especially among those with a history of frequent self mutilation. CONCLUSIONS: Dialectical behaviour therapy is superior to usual treatment in reducing high-risk behaviours in patients with BPD. PMID- 12562742 TI - Mood stabilisers plus risperidone or placebo in the treatment of acute mania. International, double-blind, randomised controlled trial. AB - BACKGROUND: Few double-blind trials have examined the efficacy of a combination of a mood stabiliser and an atypical antipsychotic in acute mania. AIMS: To determine the efficacy of risperidone in combination with a mood stabiliser in acute mania. METHOD: Patients taking a mood stabiliser were randomised to 3 weeks' treatment with risperidone (n=75) or placebo (n=76). RESULTS: Young Mania Rating Scale (YMRS) scores improved rapidly with significantly greater reductions at week 1 in the risperidone group compared with the placebo group. At end-point YMRS scores decreased by 14.5 and 10.3 points in the risperidone and placebo groups, respectively. Significant improvements v. placebo (P<0.05) were noted in the risperidone group on several other clinically meaningful measures. Additionally, a post hoc analysis excluding carbamazepine-treated patients (plasma concentrations of risperidone active moiety were 40% lower in this group) revealed significantly greater reductions (P=0.047) in YMRS scores in the risperidone group than in the placebo group. Incidence of adverse events was similar in both groups. CONCLUSIONS: Risperidone is superior to placebo when used in combination with lithium or divalproex in acute mania. PMID- 12562743 TI - Status of bipolar disorder research. Bibliometric study. AB - BACKGROUND: Bibliometric research has used publication or funding databases to compare the amount of research activity on different illnesses. Only one study has examined bipolar disorder and schizophrenia in this way, and it was restricted to one database. AIMS: The primary aim is to compare levels of research activity in bipolar disorder and schizophrenia. Secondary aims are to examine how research activity on the disorders varies over time and across scientific fields. METHOD: The numbers of publications, projects, journals and funding awards on bipolar disorder and schizophrenia were extracted from nine computer databases to compare research activity on the two conditions. RESULTS: Ratios (bipolar disorder: schizophrenia) ranged from 1:1.3 for the number of research funding awards to 1:7.6 for the number of clinical trials. CONCLUSIONS: There is a relative dearth of research activity on bipolar disorder compared with schizophrenia. PMID- 12562744 TI - Hospital doctors' management of psychological problems - Mayou & Smith revisited. AB - BACKGROUND: In 1986 the British Journal of Psychiatry published a study of physicians' and surgeons' management of psychological problems, later cited as evidence of barriers limiting liaison psychiatry. AIMS: To repeat the study after 16 years of liaison psychiatry. METHOD: Anonymous, confidential questionnaires were distributed to doctors at St George's Hospital, London, replicating the original study. RESULTS: Most of the 225 respondents believed that psychological factors could influence physical prognosis and should be routinely assessed, with greater sense of responsibility for overdoses and dying patients. Most respondents found emotional assessment impractical. Although 78% wanted more psychiatric input, referrals were avoided because of 'stigmatisation'. Men were more likely than women to hold pejorative views, but outcomes no longer varied with seniority or specialty. CONCLUSIONS: Compared with 1986, hospital doctors appear more aware of the psychological needs of patients. PMID- 12562745 TI - Flashbacks and post-traumatic stress disorder: the genesis of a 20th-century diagnosis. AB - BACKGROUND: It has been argued that post-traumatic stress disorder (PTSD) is a timeless condition, which existed before it was codified in modern diagnostic classifications but was described by different names such as 'railway spine' and 'shellshock'. Others have suggested that PTSD is a novel presentation that has resulted from a modern interaction between trauma and culture. AIMS: To test whether one core symptom of PTSD, the flashback, has altered in prevalence over time in soldiers subjected to the intense stress of combat. METHOD: Random selections were made of UK servicemen who had fought in wars from 1854 onwards and who had been awarded war pensions for post-combat disorders. These were studied to evaluate the incidence of flashbacks in defined, at-risk populations. RESULTS: The incidence of flashbacks was significantly greater in the most recent cohort, veterans of the 1991 Persian Gulf War; flashbacks were conspicuous by their absence in ex-servicemen from the Boer War and the First and Second World Wars. CONCLUSIONS: Although this study raises questions about changing interpretations of post-traumatic illness, it supports the hypothesis that some of the characteristics of PTSD are culture-bound. Earlier conflicts showed a greater emphasis on somatic symptoms. PMID- 12562746 TI - Influence of cohort effects on patterns of suicide in England and Wales, 1950 1999. AB - BACKGROUND: Age- and gender-specific suicide rates in England and Wales have changed considerably since 1950. AIMS: To assess whether cohort effects underlie some of these changes. METHOD: Graphical displays to assess age-period-cohort effects on suicide for the period 1950-1999. RESULTS: Successive male birth cohorts born after 1940 carried with them, as they aged, a greater risk of suicide than their predecessors although this effect diminished for the 1975 and 1980 birth cohorts. There was less clear evidence of any increased risk of suicide in post-war female birth cohorts. CONCLUSIONS: Succeeding generations of males born in the post-war years have experienced increasing rates of suicide at all ages, an observation in keeping with patterns seen in other countries. If these trends continue into middle- and old-age they will lead to a great increase in overall male suicide rates. PMID- 12562747 TI - Risks of combination neuroleptic treatment. PMID- 12562748 TI - Antipsychotics, HERG and sudden death. PMID- 12562749 TI - Decision-making and euthanasia. PMID- 12562750 TI - More to social capital than Putnam. PMID- 12562752 TI - Retention in psychiatry. PMID- 12562753 TI - Seasonal affective disorder. PMID- 12562754 TI - The proton transfer step catalyzed by yeast pyruvate kinase. AB - The nature of the proton donor to the C-3 of the enolate of pyruvate, the intermediate in the reaction catalyzed by yeast pyruvate kinase, was investigated by site-directed mutagenesis and physical and kinetic analyses. Thr-298 is correctly located to function as the proton donor. T298S and T298A were constructed and purified. Both mutants are catalytically active with a decrease in k(cat) and k(cat)/K(m)(,PEP). Mn(2+)-activated T298S and T298A do not exhibit homotropic kinetic cooperativity with phosphoenolpyruvate (PEP) in the absence of fructose 1,6-bisphosphate, although PEP binding to enzyme-Mn(2+) is cooperative. The pH dependence of k(cat) for T298A indicates the loss of pK(a)(,2) = 6.4-6.9. Thr-298 affects the ionization (pK(a) approximately 6.5) responsible for modulation of k(cat). Fluorescence studies show altered dissociation constants of ligands to each enzyme complex upon Thr-298 mutations. The rates of the phosphoryl transfer and proton transfer steps in the pyruvate kinase-catalyzed reaction are altered; pyruvate enolization is affected to a greater extent. Proton inventory studies demonstrate solvent isotope effects on k(cat) and k(cat)/K(m)(,PEP). Fractionation factors are metal-dependent and significantly <1. The data suggest that a water molecule in a water channel is the direct proton donor to enolpyruvate and that Thr-298 affects a late step in catalysis. PMID- 12562755 TI - Homologous binding sites in yeast isocitrate dehydrogenase for cofactor (NAD+) and allosteric activator (AMP). AB - Yeast NAD(+)-specific isocitrate dehydrogenase (IDH) is an allosterically regulated octameric enzyme composed of two types of homologous subunits designated IDH1 and IDH2. Based on sequence comparisons and structural models, both subunits are predicted to have adenine nucleotide binding sites. This was tested by alanine replacement of residues in putative sites in each subunit. Targets included adjacent aspartate/isoleucine residues implicated as important for determining cofactor specificity in related dehydrogenases and a residue in each IDH subunit in a position occupied by histidine in other cofactor binding sites. The primary kinetic effects of D286A/I287A and of H281A replacements in IDH2 were found to be a dramatic reduction in apparent affinity of the holoenzyme for NAD(+) and a concomitant reduction in V(max). Ligand binding assays also showed that the H281A mutant enzyme fails to bind NAD(+) under conditions that are saturating for the wild-type enzyme. In contrast, the primary effect of corresponding D279A/D280A and of R274A replacements in IDH1 is a reduction in holoenzyme binding of AMP, with concomitant alterations in kinetic and isocitrate binding properties normally associated with activation by this allosteric effector. These results suggest that the nucleotide cofactor binding site is primarily contributed by the IDH2 subunit, whereas the homologous nucleotide binding site in IDH1 has evolved for regulatory binding of AMP. These results are consistent with previous studies demonstrating that the catalytic isocitrate binding sites are comprised of residues primarily contributed by IDH2, whereas sites for regulatory binding of isocitrate are contributed by analogous residues of IDH1. In this study, we also demonstrate that a prerequisite for holoenzyme binding of NAD(+) is binding of isocitrate/Mg(2+) at the IDH2 catalytic site. This is comparable to the dependence of AMP binding upon binding of isocitrate at the IDH1 regulatory site. PMID- 12562756 TI - Sip2, an N-myristoylated beta subunit of Snf1 kinase, regulates aging in Saccharomyces cerevisiae by affecting cellular histone kinase activity, recombination at rDNA loci, and silencing. AB - Saccharomyces cerevisiae has evolved a number of mechanisms for sensing glucose. In the present study we examine the mechanism by which one of these pathways, involving Snf1, regulates cellular aging. Snf1 is a heterotrimer composed of a catalytic alpha subunit (Snf1p) that phosphorylates target proteins at Ser/Thr residues, an activating gamma subunit (Snf4p), and a beta subunit (Sip1p, Sip2p, or Gal83). We previously showed that forced expression of Snf1p or loss of Sip2p, but not the other beta subunits, causes accelerated aging, while removal of Snf4p extends life span (Ashrafi, K., Lin, S. S., Manchester, J. K., and Gordon, J. I. (2000) Genes Dev. 14, 1872-1885). We now demonstrate that in wild type cells, there is an age-associated shift in Sip2p from the plasma membrane to the cytoplasm, a prominent redistribution of Snf4p from the plasma membrane to the nucleus, a modest increase in nuclear Snf1p, and a concomitant increase in cellular Snf1 histone H3 kinase activity. Covalent attachment of myristate to the N-terminal Gly of Sip2p is essential for normal cellular life span. When plasma membrane association of Sip2p is abolished by a mutation that blocks its N myristoylation, Snf4p is shifted to the nucleus. Rapidly aging sip2 Delta cells have higher levels of histone H3 kinase activity than their generation-matched isogenic wild type counterparts. Increased Snf1 activity is associated with augmented recombination at rDNA loci, plus desilencing at sites affected by Snf1 catalyzed Ser(10) phosphorylation of histone H3 (the INO1 promoter plus targets of the transcription factor Adr1p). The rapid-aging phenotype of sip2 Delta cells is fully rescued by blocking recombination at rDNA loci with a fob1 Delta allele; rescue is not accompanied by amelioration of an age-associated shift toward gluconeogenesis and glucose storage. Together, these findings suggest that Sip2p acts as a negative regulator of nuclear Snf1 activity in young cells by sequestering its activating gamma subunit at the plasma membrane and that loss of Sip2p from the plasma membrane to the cytoplasm in aging cells facilities Snf4p entry into the nucleus so that Snf1 can modify chromatin structure. PMID- 12562757 TI - A eukaryotic type serine/threonine kinase and phosphatase in Streptococcus agalactiae reversibly phosphorylate an inorganic pyrophosphatase and affect growth, cell segregation, and virulence. AB - Protein phosphorylation is essential for the regulation of cell growth, division, and differentiation in both prokaryotes and eukaryotes. Signal transduction in prokaryotes was previously thought to occur primarily by histidine kinases, involved in two-component signaling pathways. Lately, bacterial homologues of eukaryotic-type serine/threonine kinases and phosphatases have been found to be necessary for cellular functions such as growth, differentiation, pathogenicity, and secondary metabolism. The Gram-positive bacteria Streptococcus agalactiae (group B streptococci, GBS) is an important human pathogen. We have identified and characterized a eukaryotic-type serine/threonine protein kinase (Stk1) and its cognate phosphatase (Stp1) in GBS. Biochemical assays revealed that Stk1 has kinase activity and localizes to the membrane and that Stp1 is a soluble protein with manganese-dependent phosphatase activity on Stk1. Mutations in these genes exhibited pleiotropic effects on growth, virulence, and cell segregation of GBS. Complementation of these mutations restored the wild type phenotype linking these genes to the regulation of various cellular processes in GBS. In vitro phosphorylation of cell extracts from wild type and mutant strains revealed that Stk1 is essential for phosphorylation of six GBS proteins. We have identified the predominant endogenous substrate of both Stk1 and Stp1 as a manganese-dependent inorganic pyrophosphatase (PpaC) by liquid chromatography/tandem mass spectrometry. These results suggest that these eukaryotic-type enzymes regulate pyrophosphatase activity and other cellular functions of S. agalactiae. PMID- 12562758 TI - Maize C4 NADP-malic enzyme. Expression in Escherichia coli and characterization of site-directed mutants at the putative nucleoside-binding sites. AB - Malic enzymes catalyze the oxidative decarboxylation of l-malate to yield pyruvate, CO(2), and NAD(P)H in the presence of a bivalent metal ion. In plants, different isoforms of the NADP-malic enzyme (NADP-ME) are involved in a wide range of metabolic pathways. The C(4)-specific NADP-ME has evolved from C(3)-type malic enzymes to represent a unique and specialized form of NADP-ME as indicated by its particular kinetic and regulatory properties. In the present study, the mature C(4)-specific NADP-ME of maize was expressed in Escherichia coli. The recombinant enzyme has essentially the same physicochemical properties and K(m) for the substrates as those of the naturally occurring NADP-ME previously characterized. However, the k(cat) was almost 7-fold higher, which may suggest that the previously purified enzyme from maize leaves was partially inactive. The recombinant NADP-ME also has a very low intrinsic NAD-dependent activity. Five mutants of NADP-ME at the postulated putative NADP-binding site(s) (Gsite5V, Gsite2V, A392G, A387G, and R237L) were constructed by site-directed mutagenesis and purified to homogeneity. The participation of these residues in substrate binding and/or the catalytic reaction was inferred by kinetic measurements and circular dichroism and intrinsic fluorescence spectra. The results obtained were compared with a predicted three-dimensional model of maize C(4) NADP-ME based on crystallographic studies of related animal NAD(P)-MEs. The data presented here represent the first prokaryotic expression of a plant NADP-ME and reveals valuable insight regarding the participation of the mutated amino acids in the binding of substrates and/or catalysis. PMID- 12562759 TI - Characterization of cyclopropane fatty-acid synthase from Sterculia foetida. AB - Cyclopropane synthase from Sterculia foetida developing seeds catalyzes the addition of a methylene group from S-adenosylmethionine to the cis double bond of oleic acid (Bao, X., Katz, S., Pollard, M., and Ohlrogge, J. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 7172-7177). To understand this enzyme better, differential expression in leaf and seed tissues, protein properties, and substrate preferences of plant cyclopropane synthase were investigated. Immunoblot analysis with antibodies raised to recombinant S. foetida cyclopropane synthase (SfCPA-FAS) revealed that SfCPA-FAS is expressed in S. foetida seeds, but not in leaves, and is a membrane protein localized to microsomal fractions. Transformed tobacco cells expressing SfCPA-FAS were labeled in vivo with L [methyl-(14)C]methionine and assayed in vitro with S-adenosyl-L-[methyl (14)C]methionine. These kinetic experiments demonstrated that dihydrosterculate was synthesized from oleic acid esterified at the sn-1 position of phosphatidylcholine (PC). Furthermore, analysis of acyl chains at sn-1 and sn-2 positions that accumulated in PC from S. foetida developing seeds and from tobacco cells expressing SfCPA-FAS also demonstrated that greater than 90% of dihydrosterculate was esterified to the sn-1 position. Thus, we conclude that SfCPA-FAS is a microsomal localized membrane protein that catalyzes the addition of methylene groups derived from S-adenosyl-L-methionine across the double bond of oleic acid esterified to the sn-1 position of PC. A survey of plant and bacterial genomes for sequences related to SfCPA-FAS indicated that a peptide domain with a putative flavin-binding site is either fused to the methyltransferase domain of the plant protein or is often found encoded by a gene adjacent to a bacterial cyclopropane synthase gene. PMID- 12562761 TI - Adenylate kinase 1 deficiency induces molecular and structural adaptations to support muscle energy metabolism. AB - Genetic ablation of adenylate kinase 1 (AK1), a member of the AK family of phosphotransfer enzymes, disturbs muscle energetic economy and decreases tolerance to metabolic stress, despite rearrangements in alternative high energy phosphoryl transfer pathways. To define the mechanisms of this adaptive response, soleus and gastrocnemius muscles from AK1(-/-) mice were characterized by cDNA array profiling, Western blot and ultrastructural analysis. We demonstrate that AK1 deficiency induces fiber-type specific variation in groups of transcripts involved in glycolysis and mitochondrial metabolism and in gene products defining structural and myogenic events. This was associated with increased phosphotransfer capacities of the glycolytic enzymes pyruvate kinase and 3 phosphoglycerate kinase. Moreover, in AK1(-/-) mice, fast-twitch gastrocnemius, but not slow-twitch soleus, had an increase in adenine nucleotide translocator (ANT) and mitochondrial creatine kinase protein, along with a doubling of the intermyofibrillar mitochondrial volume. These results provide molecular evidence for wide-scale remodeling in AK1-deficient muscles aimed at preservation of efficient energetic communication between ATP producing and utilizing cellular sites. PMID- 12562760 TI - Ure2, a prion precursor with homology to glutathione S-transferase, protects Saccharomyces cerevisiae cells from heavy metal ion and oxidant toxicity. AB - Ure2, the protein that negatively regulates GATA factor (Gln3, Gat1)-mediated transcription in Saccharomyces cerevisiae, possesses prion-like characteristics. Identification of metabolic and environmental factors that influence prion formation as well as any activities that prions or prion precursors may possess are important to understanding them and developing treatment strategies for the diseases in which they participate. Ure2 exhibits primary sequence and three dimensional homologies to known glutathione S-transferases. However, multiple attempts over nearly 2 decades to demonstrate Ure2-mediated S-transferase activity have been unsuccessful, leading to the possibility that Ure2 may well not participate in glutathionation reactions. Here we show that Ure2 is required for detoxification of glutathione S-transferase substrates and cellular oxidants. ure2 Delta mutants are hypersensitive to cadmium and nickel ions and hydrogen peroxide. They are only slightly hypersensitive to diamide, which is nitrogen source-dependent, and minimally if at all hypersensitive to 1-chloro-2,4 dinitrobenzene, the most commonly used substrate for glutathione S-transferase enzyme assays. Therefore, Ure2 shares not only structural homology with various glutathione S-transferases, but ure2 mutations possess the same phenotypes as mutations in known S. cerevisiae and Schizosaccharomyces pombe glutathione S transferase genes. These findings are consistent with Ure2 serving as a glutathione S-transferase in S. cerevisiae. PMID- 12562762 TI - Localization and function of the yeast multidrug transporter Tpo1p. AB - In Saccharomyces cerevisiae four transporters, Tpo1p-Tpo4p, all members of the major facilitator superfamily, have been shown to confer resistance to polyamines. It was suggested that they act by pumping their respective substrate into the lumen of the vacuole depending on the proton gradient generated by the V ATPase. Using sucrose gradient ultracentrifugation we found that an hemagglutinin (HA)-tagged Tpo1p as well as its HA-tagged Tpo2p-4p homologues co-localize with plasma membrane markers. Because the HA-tagged Tpo1p carrier protein proved to be functional in conferring resistance to polyamines in TPO1 knockouts, a function of Tpo1p in transport of polyamines across the plasma membrane seemed to be likely. The polyamine transport activity of wild type cells was compared with the respective activity of a TPO1 knockout strain. The results obtained strongly suggest that Tpo1p is a plasma membrane-bound exporter, involved in the detoxification of excess spermidine in yeast. When studying polyamine transport of wild type cells, we furthermore found that S. cerevisiae is excreting putrescine during the fermentative growth phase. PMID- 12562763 TI - Rotational on-off switching of a hybrid membrane sensor kinase Tar-ArcB in Escherichia coli. AB - Signal transduction in biological systems typically involves receptor proteins that possess an extracytosolic sensory domain connected to a cytosolic catalytic domain. Relatively little is known about the mechanism by which the signal is transmitted from the sensory site to the catalytic site. At least in the case of Tar (methyl-accepting chemotaxis protein for sensing aspartate) of Escherichia coli, vertical piston-like displacements of one transmembrane segment relative to the other within the monomer induced by ligand binding has been shown to modulate the catalytic activity of the cytosolic domain. The ArcB sensor kinase of E. coli is a transmembrane protein without a significant periplasmic domain. Here, we explore how the signal is conveyed to the catalytic site by analyzing the property of various Tar-ArcB hybrids. Our results suggest that, in contrast to the piston-like displacement that operates in Tar, the catalytic activity of ArcB is set by altering the orientation of the cytosolic domain of one monomer relative to the other in the homodimer. Thus, ArcB represents a distinct family of membrane receptor proteins whose catalytic activity is determined by rotational movements of the cytosolic domain. PMID- 12562764 TI - In vivo cloning and characterization of a new growth suppressor protein TOE1 as a direct target gene of Egr1. AB - Egr1, an immediate early transcription factor, responds to diverse stimuli and affects gene transcription to accomplish its biological effects. One important effect of Egr1 expression is to decrease the growth and tumorigenic potential of several tumor cell types. To identify important Egr1 target genes, we have adapted a methodology involving formaldehyde-induced protein-DNA cross-linking, chromatin immunoprecipitation, and multiplex PCR. Using this approach, we report the cloning of a new Egr1 target gene that is able to account, at least in part, for the growth inhibitory activity of Egr1. We have named this new protein TOE1 for target of Egr1. PMID- 12562765 TI - Ataxia telangiectasia mutated proteins, MAPKs, and RSK2 are involved in the phosphorylation of STAT3. AB - Phosphorylation at Ser(727) is known to be required for complete activation of STAT3 by diverse stimuli including UV irradiation, but the kinase(s) responsible for phosphorylating STAT3 (Ser(727)) is still not well discerned. In the present study, we observed that activation of ATM is required for a UVA-stimulated increase in Ser(727) phosphorylation of STAT3 as well as in activation and phosphorylation of p90 ribosomal protein S6 kinases (RSKs). Moreover, UVA stimulated activation of upstream kinases, such as c-Jun N-terminal kinases (JNKs) and ERKs, involved in mediating phosphorylation of RSKs and STAT3 was defective or delayed in ATM-deficient cells. Furthermore, we provide evidence that RSK2-deficient cells were defective for UV-induced Ser(727) phosphorylation of STAT3, and the defect was restored after ectopic expression of transfected full-length RSK2. In vitro experiments showed that active RSK2 and JNK1 induce the phosphorylation of STAT3 precipitates from immunoprecipitation but not from glutathione S-transferase (GST) pull-down. Interestingly, the GST fusion STAT3 proteins mixed together with STAT3 immunoprecipitates can be phosphorylated by JNK. However, the in vitro phosphorylation of STAT3 was reduced by the GST-STAT3 beta protein, a dominant negative form of STAT3. Taken together, our results demonstrate that the STAT3 phosphorylation at Ser(727) is triggered by active RSK2 or JNK1 in the presence of a downstream kinase or a cofactor, and thereby the intracellular phosphorylation process is stimulated through a signaling pathway involving ATM, MAPKs, RSK2, and an as yet unidentified kinase or cofactor. Additionally, RSK2-mediated phosphorylation of STAT3 (Ser(727)) was further determined to be required for basal and UVA-stimulated STAT3 transcriptional activities. PMID- 12562766 TI - Subunit exchange demonstrates a differential chaperone activity of calf alpha crystallin toward beta LOW- and individual gamma-crystallins. AB - The chaperone activity of native alpha-crystallins toward beta(LOW)- and various gamma-crystallins at the onset of their denaturation, 60 and 66 degrees C, respectively, was studied at high and low crystallin concentrations using small angle x-ray scattering (SAXS) and fluorescence energy transfer (FRET). The crystallins were from calf lenses except for one recombinant human gamma S. SAXS data demonstrated an irreversible doubling in molecular weight and a corresponding increase in size of alpha-crystallins at temperatures above 60 degrees C. Further increase is observed at 66 degrees C. More subtle conformational changes accompanied the increase in size as shown by changes in environments around tryptophan and cysteine residues. These alpha-crystallin temperature-induced modifications were found necessary to allow for the association with beta(LOW)- and gamma-crystallins to occur. FRET experiments using IAEDANS (iodoacetylaminoethylaminonaphthalene sulfonic acid)- and IAF (iodoacetamidofluorescein)-labeled subunits showed that the heat-modified alpha crystallins retained their ability to exchange subunits and that, at 37 degrees C, the rate of exchange was increased depending upon the temperature of incubation, 60 or 66 degrees C. Association with beta(LOW)- (60 degrees C) or various gamma-crystallins (66 degrees C) resulted at 37 degrees C in decreased subunit exchange in proportion to bound ligands. Therefore, beta(LOW)- and gamma crystallins were compared for their capacity to associate with alpha-crystallins and inhibit subunit exchange. Quite unexpectedly for a highly conserved protein family, differences were observed between the individual gamma-crystallin family members. The strongest effect was observed for gamma S, followed by h gamma Srec, gamma E, gamma A-F, gamma D, gamma B. Moreover, fluorescence properties of alpha crystallins in the presence of bound beta(LOW)-and gamma-crystallins indicated that the formation of beta(LOW)/alpha- or gamma/alpha-crystallin complexes involved various binding sites. The changes in subunit exchange associated with the chaperone properties of alpha-crystallins toward the other lens crystallins demonstrate the dynamic character of the heat-activated alpha-crystallin structure. PMID- 12562767 TI - Campylobacter jejuni binds intestinal H(O) antigen (Fuc alpha 1, 2Gal beta 1, 4GlcNAc), and fucosyloligosaccharides of human milk inhibit its binding and infection. AB - The most common cause of infant mortality is diarrhea; the most common cause of bacterial diarrhea is Campylobacter jejuni, which is also the primary cause of motor neuron paralysis. The first step in campylobacter pathogenesis is adherence to intestinal mucosa. We found that such binding was inhibited in vitro by human milk and, with high avidity, by alpha1,2-fucosylated carbohydrate moieties containing the H(O) blood group epitope (Fuc alpha 1,2Gal beta 1,4GlcNAc em leader ). In studies on the mechanism of adherence, campylobacter, which normally does not bind to Chinese hamster ovary cells, bound avidly when the cells were transfected with a human alpha1,2-fucosyltransferase gene that caused overexpression of H-2 antigen; binding was specifically inhibited by H-2 ligands (lectins Ulex europaeus and Lotus tetragonolobus and H-2 monoclonal antibody), H 2 mimetics, and human milk oligosaccharides. Human milk oligosaccharides inhibited campylobacter colonization of mice in vivo and human intestinal mucosa ex vivo. Campylobacter colonization of nursing mouse pups was inhibited if their dams had been transfected with a human alpha1,2-fucosyltransferase gene that caused expression of H(O) antigen in milk. We conclude that campylobacter binding to intestinal H-2 antigen is essential for infection. Milk fucosyloligosaccharides and specific fucosyl alpha1,2-linked molecules inhibit this binding and may represent a novel class of antimicrobial agents. PMID- 12562768 TI - Molecular evolution by change of function. Alkaloid-specific homospermidine synthase retained all properties of deoxyhypusine synthase except binding the eIF5A precursor protein. AB - Deoxyhypusine synthase participates in the post-translational activation of the eukaryotic initiation factor 5A (eIF5A). The enzyme transfers the aminobutyl moiety of spermidine to a specific lysine residue in the eIF5A precursor protein, i.e. eIF5A(lys). Homospermidine synthase catalyzes an analogous reaction but uses putrescine instead of eIF5A(lys) as substrate yielding the rare polyamine homospermidine as product. Homospermidine is an essential precursor in the biosynthesis of pyrrolizidine alkaloids, an important class of plant defense compounds against herbivores. Sequence comparisons of the two enzymes indicate an evolutionary origin of homospermidine synthase from ubiquitous deoxyhypusine synthase. The two recombinant enzymes from Senecio vernalis were purified, and their properties were compared. Protein-protein binding and kinetic substrate competition studies confirmed that homospermidine synthase, in comparison to deoxyhypusine synthase, lost the ability to bind the eIF5A(lys) to its surface. The two enzymes show the same unique substrate specificities, catalyze the aminobutylation of putrescine with the same specific activities, and exhibit almost identical Michaelis kinetics. In conclusion, homospermidine synthase behaves like a deoxyhypusine synthase that lost its major function (aminobutylation of eIF5A precursor protein) but retained unaltered its side activity (aminobutylation of putrescine). It is suggested as having evolved from deoxyhypusine synthase by gene duplication and being recruited for a new function. PMID- 12562769 TI - Procyclic Trypanosoma brucei do not use Krebs cycle activity for energy generation. AB - The importance of a functional Krebs cycle for energy generation in the procyclic stage of Trypanosoma brucei was investigated under physiological conditions during logarithmic phase growth of a pleomorphic parasite strain. Wild type procyclic cells and mutants with targeted deletion of the gene coding for aconitase were derived by synchronous in vitro differentiation from wild type and mutant (Delta aco::NEO/Delta aco::HYG) bloodstream stage parasites, respectively, where aconitase is not expressed and is dispensable. No differences in intracellular levels of glycolytic and Krebs cycle intermediates were found in procyclic wild type and mutant cells, except for citrate that accumulated up to 90-fold in the mutants, confirming the absence of aconitase activity. Surprisingly, deletion of aconitase did not change differentiation nor the growth rate or the intracellular ATP/ADP ratio in those cells. Metabolic studies using radioactively labeled substrates and NMR analysis demonstrated that glucose and proline were not degraded via the Krebs cycle to CO(2). Instead, glucose was degraded to acetate, succinate, and alanine, whereas proline was degraded to succinate. Importantly, there was absolutely no difference in the metabolic products released by wild type and aconitase knockout parasites, and both were for survival strictly dependent on respiration via the mitochondrial electron transport chain. Hence, although the Krebs cycle enzymes are present, procyclic T. brucei do not use Krebs cycle activity for energy generation, but the mitochondrial respiratory chain is essential for survival and growth. We therefore propose a revised model of the energy metabolism of procyclic T. brucei. PMID- 12562770 TI - Structure of human carnitine acetyltransferase. Molecular basis for fatty acyl transfer. AB - Carnitine acyltransferases are a family of ubiquitous enzymes that play a pivotal role in cellular energy metabolism. We report here the x-ray structure of human carnitine acetyltransferase to a 1.6-A resolution. This structure reveals a monomeric protein of two equally sized alpha/beta domains. Each domain is shown to have a partially similar fold to other known but oligomeric enzymes that are also involved in group-transfer reactions. The unique monomeric arrangement of the two domains constitutes a central narrow active site tunnel, indicating a likely universal feature for all members of the carnitine acyltransferase family. Superimposition of the substrate complex of a related protein, dihydrolipoyl trans-acetylase, reveals that both substrates localize to the active site tunnel of human carnitine acetyltransferase, suggesting the location of the ligand binding sites for carnitine and coenzyme A. Most significantly, this structure provides critical insights into the molecular basis for fatty acyl chain transfer and a possible common mechanism among a wide range of acyltransferases utilizing a catalytic dyad. PMID- 12562771 TI - Identification and characterization of ADAMTS-20 defines a novel subfamily of metalloproteinases-disintegrins with multiple thrombospondin-1 repeats and a unique GON domain. AB - We have cloned a mouse brain cDNA encoding a new protein of the ADAMTS family (a disintegrin and metalloproteinase domain, with thrombospondin type-1 repeats), which has been called ADAMTS-20. This protein shows a domain organization similar to that described for other ADAMTSs including signal sequence, propeptide, metalloproteinase domain, disintegrin domain, central TS-1 motif, cysteine-rich region, and C-terminal TS module. However, this last module is more complex than that of other ADAMTSs, being composed of a total of 14 repeats. The structural complexity of ADAMTS-20 is further increased by the presence of an additional domain 200 residues long and located immediately adjacent to the TS module. This domain has been tentatively called GON domain and can also be recognized in some ADAMTSs such as gon-1 from Caenorhabditis elegans and human and mouse ADAMTS-9. The presence of this domain is a hallmark of a novel subfamily of structurally and evolutionarily related ADAMTSs, called GON-ADAMTSs. Expression analysis demonstrated that ADAMTS-20 transcripts can be detected at low levels in several human and mouse tissues, especially in testis. This gene is also overexpressed in some human malignant tumors, including brain, colon, and breast carcinomas. Western blot analysis using polyclonal antibodies raised against recombinant ADAMTS-20 produced in Escherichia coli showed the presence of a 70-kDa band in mouse brain and testis extracts. This recombinant ADAMTS-20 hydrolyzed a synthetic peptide used for assaying matrix metalloproteinases. These data suggest that this novel enzyme may play a role in the tissue remodeling process occurring in both normal and pathological conditions. PMID- 12562772 TI - Ubiquitin-independent mechanisms of mouse ornithine decarboxylase degradation are conserved between mammalian and fungal cells. AB - The polyamine biosynthetic enzyme ornithine decarboxylase (ODC) is degraded by the 26 S proteasome via a ubiquitin-independent pathway in mammalian cells. Its degradation is greatly accelerated by association with the polyamine-induced regulatory protein antizyme 1 (AZ1). Mouse ODC (mODC) that is expressed in the yeast Saccharomyces cerevisiae is also rapidly degraded by the proteasome of that organism. We have now carried out in vivo and in vitro studies to determine whether S. cerevisiae proteasomes recognize mODC degradation signals. Mutations of mODC that stabilized the protein in animal cells also did so in the fungus. Moreover, the mODC degradation signal was able to destabilize a GFP or Ura3 reporter in GFP-mODC and Ura3-mODC fusion proteins. Co-expression of AZ1 accelerated mODC degradation 2-3-fold in yeast cells. The degradation of both mODC and the endogenous yeast ODC (yODC) was unaffected in S. cerevisiae mutants with various defects in ubiquitin metabolism, and ubiquitinylated forms of mODC were not detected in yeast cells. In addition, recombinant mODC was degraded in an ATP-dependent manner by affinity-purified yeast 26 S proteasomes in the absence of ubiquitin. Degradation by purified yeast proteasomes was sensitive to mutations that stabilized mODC in vivo, but was not accelerated by recombinant AZ1. These studies demonstrate that cell constituents required for mODC degradation are conserved between animals and fungi, and that both mammalian and fungal ODC are subject to proteasome-mediated proteolysis by ubiquitin independent mechanisms. PMID- 12562773 TI - NMR study of mersacidin and lipid II interaction in dodecylphosphocholine micelles. Conformational changes are a key to antimicrobial activity. AB - Mersacidin belongs to the type B lantibiotics (lanthionine-containing antibiotics) that contain post-translationally modified amino acids and cyclic ring structures. It targets the cell wall precursor lipid II and thereby inhibits cell wall synthesis. In light of the emerging antibiotics resistance problem, the understanding of the antibacterial activity on a structural basis provides a key to circumvent this issue. Here we present solution NMR studies of mersacidin lipid II interaction in dodecylphosphocholine (DPC) micelles. Distinct solution structures of mersacidin were determined in three different states: in water/methanol solution and in DPC micelles with and without lipid II. The structures in various sample conditions reveal remarkable conformational changes in which the junction between Ala-12 and Abu-13 (where Abu is aminobutyric acid) effectively serves as the hinge for the opening and closure of the ring structures. The DPC micelle-bound form resembles the previously determined NMR and x-ray crystal structures of mersacidin in pure methanol but substantially deviates from the other two states in our current report. The structural changes delineate the large chemical shift perturbations observed during the course of a two-step (15)N-(1)H heteronuclear single quantum coherence titration. They also modulate the surface charge distribution of mersacidin suggesting that electrostatics play a central role in the mersacidin-lipid II interaction. The observed conformational adaptability of mersacidin might be a general feature of lipid II-interacting antibiotics/peptides. PMID- 12562774 TI - Crystal structure of an alpha 1,4-N-acetylhexosaminyltransferase (EXTL2), a member of the exostosin gene family involved in heparan sulfate biosynthesis. AB - EXTL2, an alpha1,4-N-acetylhexosaminyltransferase, catalyzes the transfer reaction of N-acetylglucosamine and N-acetylgalactosamine from the respective UDP sugars to the non-reducing end of [glucuronic acid]beta1-3[galactose]beta1-O naphthalenemethanol, an acceptor substrate analog of the natural common linker of various glycosylaminoglycans. We have solved the x-ray crystal structure of the catalytic domain of mouse EXTL2 in the apo-form and with donor substrates UDP-N acetylglucosamine and UDP-N-acetylgalactosamine. In addition, a structure of the ternary complex with UDP and the acceptor substrate analog [glucuronic acid]beta1 3[galactose]beta1-O-naphthalenemethanol has been determined. These structures reveal three highly conserved residues, Asn-243, Asp-246, and Arg-293, located at the active site. Mutation of these residues greatly decreases the activity. In the ternary complex, an interaction exists between the beta-phosphate of the UDP leaving group and the acceptor hydroxyl of the substrate that may play a functional role in catalysis. These structures represent the first structures from the exostosin gene family and provide important insight into the mechanisms of alpha1,4-N-acetylhexosaminyl transfer in heparan biosynthesis. PMID- 12562775 TI - Specific nitration at tyrosine 430 revealed by high resolution mass spectrometry as basis for redox regulation of bovine prostacyclin synthase. AB - Treatment of bovine aortic microsomes containing active prostacyclin synthase (PGI(2) synthase) with increasing concentrations of peroxynitrite (PN) up to 250 microm of PN yielded specific staining of this enzyme on Western blots with antibodies against 3-nitrotyrosine (3-NT), whereas above 500 microm PN staining of additional proteins was also observed. Following treatment of aortic microsomes with 25 microm PN, PGI(2) synthase was about half-maximally nitrated and about half-inhibited. It was then isolated by gel electrophoresis and subjected to proteolytic digestion with several proteases. Digestion with thermolysin for 24 h provided a single specific peptide that was isolated by high performance liquid chromatography and identified as a tetrapeptide Leu-Lys-Asn Tyr(3-nitro)-COOH corresponding to positions 427-430 of PGI(2) synthase. Its structure was established by precise mass determination using Fourier transform ion cyclotron resonance-nanoelectrospray mass spectrometry and Edman microsequencing and ascertained by synthesis and mass spectrometric characterization of the authentic Tyr-nitrated peptide. Complete digestion by Pronase to 3-nitrotyrosine was obtained only after 72 h, suggesting that the nitrated Tyr-430 residue may be embedded in a tight fold around the heme binding site. These results provide evidence for the specific inhibition of PGI(2) synthase by nitration at Tyr-430 that may occur already at low levels of PN as a consequence of endothelial co-generation of nitric oxide and superoxide. PMID- 12562776 TI - Mutations of Arg440 and Gly455/Gly456 oppositely change pH sensing of Na+/H+ exchanger 1. AB - To identify important amino acid residues involved in intracellular pH (pH(i)) sensing of Na(+)/H(+) exchanger 1, we produced single-residue substitution mutants in the region of the exchanger encompassing the putative 11th transmembrane segment (TM11) and its adjacent intracellular (intracellular loop (IL) 5) and extracellular loops (extracellular loop 6). Substitution of Arg(440) in IL5 with other residues except positively charged Lys caused a large shift in pH(i) dependence of (22)Na(+) uptake to an acidic side, whereas substitution of Gly(455) or Gly(456) within the highly conserved glycine-rich sequence of TM11 shifted pH(i) dependence to an alkaline side. The observed alkaline shift was larger with substitution of Gly(455) with residues with increasing sizes, suggesting the involvement of the steric effect. Interestingly, mutation of Arg(440) (R440D) abolished the ATP depletion-induced acidic shift in pH(i) dependence of (22)Na(+) uptake as well as the cytoplasmic alkalinization induced by various extracellular stimuli, whereas with that of Gly(455) (G455Q) these functions were preserved. These mutant exchangers did not alter apparent affinities for extracellular transport substrates Na(+) and H(+) and the inhibitor 5-(N-ethyl-N-isopropyl)amiloride. These results suggest that positive charge at Arg(440) is required for normal pH(i) sensing, whereas mutation-induced perturbation of the TM11 structure may be involved in the effects of Gly mutations. Thus, both Arg(440) in IL5 and Gly residues in the conserved segment of TM11 appear to constitute important elements for proper functioning of the putative "pH(i) sensor" of Na(+)/H(+) exchanger 1. PMID- 12562777 TI - Essential role of a GXXXG motif for membrane channel formation by Helicobacter pylori vacuolating toxin. AB - Helicobacter pylori secretes a toxin, VacA, that can form anion-selective membrane channels. Within a unique amino-terminal hydrophobic region of VacA, there are three tandem GXXXG motifs (defined by glycines at positions 14, 18, 22, and 26), which are characteristic of transmembrane dimerization sequences. The goals of the current study were to investigate whether these GXXXG motifs are required for membrane channel formation and cytotoxicity and to clarify the role of membrane channel formation in the biological activity of VacA. Six different alanine substitution mutations (P9A, G13A, G14A, G18A, G22A, and G26A) were introduced into the unique hydrophobic region located near the amino terminus of VacA. The effects of these mutations were first analyzed using the TOXCAT system, which permits the study of transmembrane oligomerization of proteins in a natural membrane environment. None of the mutations altered the capacity of ToxR-VacA maltose-binding protein fusion proteins to insert into a membrane, but G14A and G18A mutations markedly diminished the capacity of the fusion proteins to oligomerize. We then introduced the six alanine substitution mutations into the vacA chromosomal gene of H. pylori and analyzed the properties of purified mutant VacA proteins. VacA-G13A, VacA-G22A, and VacA-G26A induced vacuolation of HeLa cells, whereas VacA-P9A, VacA-G14A, and VacA-G18A did not. Subsequent experiments examined the capacity of each mutant toxin to form membrane channels. In a planar lipid bilayer assay, VacA proteins containing G13A, G22A, and G26A mutations formed anion-selective membrane channels, whereas VacA proteins containing P9A, G14A, and G18A mutations did not. Similarly, VacA-G13A, VacA-G22A, and VacA-G26A induced depolarization of HeLa cells, whereas VacA-P9A, VacA-G14A, and VacA-G18A did not. These data indicate that an intact proline residue and an intact G(14)XXXG(18) motif within the amino-terminal hydrophobic region of VacA are essential for membrane channel formation, and they also provide strong evidence that membrane channel formation is essential for VacA cytotoxicity. PMID- 12562778 TI - Syntaxin 1A regulates ENaC via domain-specific interactions. AB - The epithelial sodium channel (ENaC) is a heterotrimeric protein responsible for Na(+) absorption across the apical membranes of several absorptive epithelia. The rate of Na(+) absorption is governed in part by regulated membrane trafficking mechanisms that control the apical membrane ENaC density. Previous reports have implicated a role for the t-SNARE protein, syntaxin 1A (S1A), in the regulation of ENaC current (I(Na)). In the present study, we examine the structure-function relations influencing S1A-ENaC interactions. In vitro pull-down assays demonstrated that S1A directly interacts with the C termini of the alpha-, beta-, and gamma-ENaC subunits but not with the N terminus of any ENaC subunit. The H3 domain of S1A is the critical motif mediating S1A-ENaC binding. Functional studies in ENaC expressing Xenopus oocytes revealed that deletion of the H3 domain of co-expressed S1A eliminated its inhibition of I(Na), and acute injection of a GST-H3 fusion protein into ENaC expressing oocytes inhibited I(Na) to the same extent as S1A co-expression. In cell surface ENaC labeling experiments, reductions in plasma membrane ENaC accounted for the H3 domain inhibition of I(Na). Individually substituting C terminus-truncated alpha-, beta , or gamma-ENaC subunits for their wild-type counterparts reversed the S1A induced inhibition of I(Na), and oocytes expressing ENaC comprised of three C terminus-truncated subunits showed no S1A inhibition of I(Na). C terminus truncation or disruption of the C terminus beta-subunit PY motif increases I(Na) by interfering with ENaC endocytosis. In contrast to subunit truncation, a beta ENaC PY mutation did not relieve S1A inhibition of I(Na), suggesting that S1A does not perturb Nedd4 interactions that lead to ENaC endocytosis/degradation. This study provides support for the concept that S1A inhibits ENaC-mediated Na(+) transport by decreasing cell surface channel number via direct protein-protein interactions at the ENaC C termini. PMID- 12562779 TI - Triiodothyronine increases brain natriuretic peptide (BNP) gene transcription and amplifies endothelin-dependent BNP gene transcription and hypertrophy in neonatal rat ventricular myocytes. AB - Brain natriuretic peptide (BNP) gene expression is a well documented marker of hypertrophy in the cardiac myocyte. Triiodothyronine (T(3)), the bioactive form of thyroid hormone, triggers a unique form of hypertrophy in cardiac myocytes that accompanies the selective activation or suppression of specific gene targets. In this study, we show that the BNP gene is a target of T(3) action. BNP secretion was increased 6-fold, BNP mRNA levels 3-fold, and BNP promoter activity 3-5-fold following T(3) treatment. This was accompanied by an increase in myocyte size, sarcomeric organization, and protein synthesis. Of note, several of the responses to T(3) synergized with those to the conventional hypertrophic agonist endothelin. The response to the liganded thyroid hormone receptor (TR) was mediated by an unusual thyroid hormone response element located between -1000 and -987 relative to the transcription start site. Both TR homodimers and TR.retinoid X receptor heterodimers associated with this element in an electrophoretic mobility shift assay. Protein fragments harboring the LXXLL motifs of the coactivators GRIP1 and SRC1 or TRAP220 interacted predominantly with the TR.retinoid X receptor heterodimeric pair in a ligand-dependent fashion. Both TR homodimers and heterodimers in the unliganded state selectively associated with glutathione S-transferase-nuclear receptor corepressor fragments harboring one of three receptor interaction domains containing the sequence (I/L)XX(I/V)I. These interactions were dissociated following the addition of T(3). Collectively, these findings identify the BNP gene as a potential model for the investigation of TR dependent gene regulation in the heart. PMID- 12562780 TI - New insights into the developmental history of the bacterial cell division site. PMID- 12562782 TI - Branching of Escherichia coli cells arises from multiple sites of inert peptidoglycan. AB - Some strains of Escherichia coli defective for dacA, the gene coding for penicillin-binding protein 5, exhibit a strong branching phenotype when cell division is blocked. Since such branch formation implies a differentiation of polar caps at ectopic locations in the cell envelope, we analyzed murein segregation and observed a strong correlation between areas of inert murein and these morphological anomalies. In particular, the tips of branches exhibited the same properties as those described for polar caps of wild-type cells, i.e., the synthesis and turnover of murein were inhibited. Also, the mobility of cell envelope proteins was apparently constrained in areas with morphological defects. Polar regions of branching cells and sacculi had aberrant morphologies with a very high frequency. Of special interest was that areas of inert murein at polar caps were often split by areas of active synthesis, a situation unlike that observed in wild-type cells. These observations suggest that in dacA mutants, branches and other morphological anomalies may arise from split polar caps or by de novo generation of new poles built around inert peptidoglycan patches in the side walls of the cell. PMID- 12562781 TI - Prokaryotic development: emerging insights. PMID- 12562784 TI - YfiK from Escherichia coli promotes export of O-acetylserine and cysteine. AB - yfiK was discovered as a gene augmenting cysteine production when it was overexpressed in an industrial Escherichia coli production strain. The gene product is an integral membrane protein with about six predicted transmembrane helices; it belongs to the RhtB family of export proteins. YfiK overproduction from a plasmid leads to drastic and parallel secretion of O-acetylserine and cysteine into the medium but only when the organism possesses a serine transacetylase that is feedback insensitive to cysteine. Externally provided O acetylserine obviated this requirement for cysteine secretion both in the yfiK carrying transformant and in the wild type. A DeltayfiK mutant did not show any phenotype, and it exported O-acetylserine and cysteine when transformed with a plasmid carrying ydeD, a previously characterized, alternate O acetylserine/cysteine exporter. Since a ydeD-yfiK double mutant showed the same pattern, it appears that YfiK and YdeD act independently. The necessity for the cell to regulate the size of the internal pool of O-acetylserine via synthesis of exporter proteins could be connected to the fact that this compound (when supplied externally) inhibits growth. Overexpression of either ydeD or yfiK leads to alleviation of this inhibition paralled by increased resistance to azaserine, which is an analog of O-acetylserine. PMID- 12562783 TI - Molecular analysis of the gene encoding a novel cold-adapted chitinase (ChiB) from a marine bacterium, Alteromonas sp. strain O-7. AB - The chitinase B (ChiB) secreted by Alteromonas sp. strain O-7 was purified, and the corresponding gene (chiB) was cloned and sequenced. The open reading frame of the chiB gene encodes a protein of 850 amino acids with a calculated molecular mass of 90,223 Da. ChiB is a modular enzyme consisting of two reiterated domains and a catalytic domain belonging to chitinase family 18. The reiterated domains are composed of chitin-binding domain (ChtBD) type 3 and two fibronectin type III (Fn3)-like domains. Expression plasmids coding for ChiB or deletion derivatives thereof were constructed in Escherichia coli. Deletion analysis showed that the ChtBD of ChiB plays an important role in efficient hydrolysis of insoluble chitin. The optimum pH and temperature of ChiB were 6.0 and 30 degrees C, respectively. The enzyme showed relatively high catalysis, even at low temperatures close to 0 degrees C, and remarkable thermal lability compared to ChiA and ChiC, which are the mesophilic chitinases of the same strain. The kca)/Km value for the ChiB reaction at 10 degrees C was about 4.7 times higher than that of ChiC. These results suggest that ChiB is a cold-adapted enzyme. The RNA transcript of chiB was induced by 1% GlcNAc, and along with a rise in temperature, the RNA transcript showed a tendency to decrease. Thus, among the ChiA, ChiB, and ChiC chitinases, production of ChiB may be advantageous for the strain, allowing it to easily acquire nutrients from chitin and to survive in cold environments. PMID- 12562785 TI - In vivo assay for low-activity mutant forms of Escherichia coli ribonucleotide reductase. AB - Ribonucleotide reductase (RNR) catalyzes the essential production of deoxyribonucleotides in all living cells. In this study we have established a sensitive in vivo assay to study the activity of RNR in aerobic Escherichia coli cells. The method is based on the complementation of a chromosomally encoded nonfunctional RNR with plasmid-encoded RNR. This assay can be used to determine in vivo activity of RNR mutants with activities beyond the detection limits of traditional in vitro assays. E. coli RNR is composed of two homodimeric proteins, R1 and R2. The R2 protein contains a stable tyrosyl radical essential for the catalysis that takes place at the R1 active site. The three-dimensional structures of both proteins, phylogenetic studies, and site-directed mutagenesis experiments show that the radical is transferred from the R2 protein to the active site in the R1 protein via a radical transfer pathway composed of at least nine conserved amino acid residues. Using the new assay we determined the in vivo activity of mutants affecting the radical transfer pathway in RNR and identified some residual radical transfer activity in two mutant R2 constructs (D237N and W48Y) that had previously been classified as negative for enzyme activity. In addition, we show that the R2 mutant Y356W is completely inactive, in sharp contrast to what has previously been observed for the corresponding mutation in the mouse R2 enzyme. PMID- 12562786 TI - Interactions between phage-shock proteins in Escherichia coli. AB - Expression of the pspABCDE operon of Escherichia coli is induced upon infection by filamentous phage and by many other stress conditions, including defects in protein export. Expression of the operon requires the alternative sigma factor sigma54 and the transcriptional activator PspF. In addition, PspA plays a negative regulatory role, and the integral-membrane proteins PspB and PspC play a positive one. In this study, we investigated whether the suggested protein protein interactions implicated in this complex regulatory network can indeed be demonstrated. Antisera were raised against PspB, PspC, and PspD, which revealed, in Western blotting experiments, that PspC forms stable sodium dodecyl sulfate resistant dimers and that the hypothetical pspD gene is indeed expressed in vivo. Fractionation experiments showed that PspD localizes as a peripherally bound inner membrane protein. Cross-linking studies with intact cells revealed specific interactions of PspA with PspB and PspC, but not with PspD. Furthermore, affinity chromatography suggested that PspB could bind PspA only in the presence of PspC. These data indicate that regulation of the psp operon is mediated via protein protein interactions. PMID- 12562787 TI - CDP-2,3-Di-O-geranylgeranyl-sn-glycerol:L-serine O-archaetidyltransferase (archaetidylserine synthase) in the methanogenic archaeon Methanothermobacter thermautotrophicus. AB - CDP-2,3-di-O-geranylgeranyl-sn-glycerol:L-serine O-archaetidyltransferase (archaetidylserine synthase) activity in cell extracts of Methanothermobacter thermautotrophicus cells was characterized. The enzyme catalyzed the formation of unsaturated archaetidylserine from CDP-unsaturated archaeol and L-serine. The identity of the reaction products was confirmed by thin-layer chromatography, fast atom bombardment-mass spectrum analysis, and chemical degradation. The enzyme showed maximal activity in the presence of 10 mM Mn2+ and 1% Triton X-100. Among various synthetic substrate analogs, both enantiomers of CDP-unsaturated archaeols with ether-linked geranylgeranyl chains and CDP-saturated archaeol with ether-linked phytanyl chains were similarly active toward the archaetidylserine synthase. The activity on the ester analog of the substrate was two to three times higher than that on the corresponding ether-type substrate. The activity of D-serine with the enzyme was 30% of that observed for L-serine. A trace amount of an acid-labile, unsaturated archaetidylserine intermediate was detected in the cells by a pulse-labeling experiment. A gene (MT1027) in M. thermautotrophicus genome annotated as the gene encoding phosphatidylserine synthase was found to be homologous to Bacillus subtilis pssA but not to Escherichia coli pssA. The substrate specificity of phosphatidylserine synthase from B. subtilis was quite similar to that observed for the M. thermautotrophicus archaetidylserine synthase, while the E. coli enzyme had a strong preference for CDP-1,2-diacyl-sn glycerol. It was concluded that M. thermautotrophicus archaetidylserine synthase belongs to subclass II phosphatidylserine synthase (B. subtilis type) on the basis of not only homology but also substrate specificity and some enzymatic properties. The possibility that a gene encoding the subclass II phosphatidylserine synthase might be transferred from a bacterium to an ancestor of methanogens is discussed. PMID- 12562788 TI - Effect of intracellular pH on rotational speed of bacterial flagellar motors. AB - Weak acids such as acetate and benzoate, which partially collapse the transmembrane proton gradient, not only mediate pH taxis but also impair the motility of Escherichia coli and Salmonella at an external pH of 5.5. In this study, we examined in more detail the effect of weak acids on motility at various external pH values. A change of external pH over the range 5.0 to 7.8 hardly affected the swimming speed of E. coli cells in the absence of 34 mM potassium acetate. In contrast, the cells decreased their swimming speed significantly as external pH was shifted from pH 7.0 to 5.0 in the presence of 34 mM acetate. The total proton motive force of E. coli cells was not changed greatly by the presence of acetate. We measured the rotational rate of tethered E. coli cells as a function of external pH. Rotational speed decreased rapidly as the external pH was decreased, and at pH 5.0, the motor stopped completely. When the external pH was returned to 7.0, the motor restarted rotating at almost its original level, indicating that high intracellular proton (H+) concentration does not irreversibly abolish flagellar motor function. Both the swimming speeds and rotation rates of tethered cells of Salmonella also decreased considerably when the external pH was shifted from pH 7.0 to 5.5 in the presence of 20 mM benzoate. We propose that the increase in the intracellular proton concentration interferes with the release of protons from the torque-generating units, resulting in slowing or stopping of the motors. PMID- 12562789 TI - Analysis of residues determining specificity of Vibrio cholerae TonB1 for its receptors. AB - In gram-negative organisms, high-affinity transport of iron substrates requires energy transduction to specific outer membrane receptors by the TonB-ExbB-ExbD complex. Vibrio cholerae encodes two TonB proteins, one of which, TonB1, recognizes only a subset of V. cholerae TonB-dependent receptors and does not facilitate transport through Escherichia coli receptors. To investigate the receptor specificity exhibited by V. cholerae TonB1, chimeras were created between V. cholerae TonB1 and E. coli TonB. The activities of the chimeric TonB proteins in iron utilization assays demonstrated that the C-terminal one-third of either TonB confers the receptor specificities associated with the full-length TonB. Single-amino-acid substitutions near the C terminus of V. cholerae TonB1 were identified that allowed TonB1 to recognize E. coli receptors and at least one V. cholerae TonB2-dependent receptor. This indicates that the very C-terminal end of V. cholerae TonB1 determines receptor specificity. The regions of the TonB dependent receptors involved in specificity for a particular TonB protein were investigated in experiments involving domain switching between V. cholerae and E. coli receptors exhibiting different TonB specificities. Switching the conserved TonB box heptapeptides at the N termini of these receptors did not alter their TonB specificities. However, replacing the amino acid immediately preceding the TonB box in E. coli receptors with an aromatic residue allowed these receptors to use V. cholerae TonB1. Further, site-directed mutagenesis of the TonB box -1 residue in a V. cholerae TonB2-dependent receptor demonstrated that a large hydrophobic amino acid in this position promotes recognition of V. cholerae TonB1. These data suggest that the TonB box -1 position controls productive interactions with V. cholerae TonB1. PMID- 12562790 TI - Molecular genetic analysis of a group A Streptococcus operon encoding serum opacity factor and a novel fibronectin-binding protein, SfbX. AB - The group A Streptococcus (GAS) sof gene encodes the serum opacity factor protein, which is capable of opacifying mammalian sera and binding at least two host proteins, fibronectin and fibrinogen. The sof gene exists in approximately 50% of clinical isolates, and there is a classical association of so-called nephritogenic strains with the opacity factor-positive phenotype. In both a type emm49 strain and a type emm12 strain, the sequences upstream of the 5' end of sof and downstream of the putative terminator were determined to be nearly identical to a region in the M type 1 genome approximately 10 kb upstream of the emm1 gene. This close genetic linkage is likely reflected in the strict correlation of opacity factor phenotype with specific emm genotypes. A new fibronectin-binding protein gene, sfbX, was discovered immediately downstream of sof in emm12 and emm49 strains and in several other sof-positive strains. The sof and sfbX genes were found to be expressed on the same transcription unit, which was correlated with the putative promoter and rho-independant terminator sequences that flank these two genes. The sfbX genes from different emm types are predicted to encode approximately 650-residue surface-bound proteins sharing 89 to 92% sequence identity. SfbX residues approximately 1 to 480 are not highly similar to those of other known proteins, with the closest match being the Staphylococcus aureus coagulase protein. The remaining portions of these proteins (residues 481 to 650) contain four putative fibronectin-binding repeats highly similar to those of other streptococcal fibronectin-binding proteins and a potential LP(X)SG cell wall anchor motif. Targeted in-frame allelic-exchange mutagenesis, complementation, and heterologous-expression studies found that serum opacification is encoded by sof alone and that sfbX encodes a fibronectin-binding function. A recombinant SfbX protein was found to bind immobilized fibronectin and to partially inhibit GAS adherence to fibronectin. The sfbX gene was found to be present only in sof-positive strains, and together these genes could influence the spectrum of tissues colonized by sof-positive GAS. PMID- 12562791 TI - In vitro and in vivo functional activity of Chlamydia MurA, a UDP-N acetylglucosamine enolpyruvyl transferase involved in peptidoglycan synthesis and fosfomycin resistance. AB - Organisms of Chlamydia spp. are obligate intracellular, gram-negative bacteria with a dimorphic developmental cycle that takes place entirely within a membrane bound vacuole termed an inclusion. The chlamydial anomaly refers to the fact that cell wall-active antibiotics inhibit Chlamydia growth and peptidoglycan (PG) synthesis genes are present in the genome, yet there is no biochemical evidence for synthesis of PG. In this work, we undertook a genetics-based approach to reevaluate the chlamydial anomaly by characterizing MurA, a UDP-N acetylglucosamine enolpyruvyl transferase that catalyzes the first committed step of PG synthesis. The murA gene from Chlamydia trachomatis serovar L2 was cloned and placed under the control of the arabinose-inducible, glucose-repressible ara promoter and transformed into Escherichia coli. After transduction of a lethal DeltamurA mutation into the strain, viability of the E. coli strain became dependent upon expression of the C. trachomatis murA. DNA sequence analysis of murA from C. trachomatis predicted a cysteine-to-aspartate change in a key residue within the active site of MurA. In E. coli, the same mutation has previously been shown to cause resistance to fosfomycin, a potent antibiotic that specifically targets MurA. In vitro activity of the chlamydial MurA was resistant to high levels of fosfomycin. Growth of C. trachomatis was also resistant to fosfomycin. Moreover, fosfomycin resistance was imparted to the E. coli strain expressing the chlamydial murA. Conversion of C. trachomatis elementary bodies to reticulate bodies and cell division are correlated with expression of murA mRNA. mRNA from murB, the second enzymatic reaction in the PG pathway, was also detected during C. trachomatis infection. Our findings, as well as work from other groups, suggest that a functional PG pathway exists in Chlamydia spp. We propose that chlamydial PG is essential for progression through the developmental cycle as well as for cell division. Elucidating the existence of PG in Chlamydia spp. is of significance for the development of novel antibiotics targeting the chlamydial cell wall. PMID- 12562792 TI - QscR, a LysR-type transcriptional regulator and CbbR homolog, is involved in regulation of the serine cycle genes in Methylobacterium extorquens AM1. AB - A new gene, qscR, encoding a LysR-type transcriptional regulator that is a homolog of CbbR, has been characterized from the facultative methylotroph Methylobacterium extorquens AM1 and shown to be the major regulator of the serine cycle, the specific C1 assimilation pathway. The qscR mutant was shown to be unable to grow on C1 compounds, and it lacked the activity of serine-glyoxylate aminotransferase, a key enzyme of the serine cycle. Activities of other serine cycle enzymes were decreased during growth on C1 compounds compared to the activities found in wild-type M. extorquens AM1. Promoter fusion assays, as well as reverse transcription-PCR assays, have indicated that the serine cycle genes belong to three separate transcriptional units, sga-hpr-mtdA-fch, mtkA-mtkB-ppc mcl, and gly. Gel retardation assays involving the purified QscR have demonstrated the specific binding of QscR to the DNA regions upstream of sga, mtkA, gly, and qscR. We conclude that QscR acts as a positive transcriptional regulator of most of the serine cycle enzymes and also as an autorepressor. PMID- 12562793 TI - Roles of NhaA, NhaB, and NhaD Na+/H+ antiporters in survival of Vibrio cholerae in a saline environment. AB - Vibrio cholerae, the causative agent of cholera, is a normal inhabitant of aquatic environments, where it survives in a wide range of conditions of pH and salinity. In this work, we investigated the role of three Na+/H+ antiporters on the survival of V. cholerae in a saline environment. We have previously cloned the Vc-nhaA gene encoding the V. cholerae homolog of Escherichia coli. Here we identified two additional antiporter genes, designated Vc-nhaB and Vc-nhaD, encoding two putative proteins of 530 and 477 residues, respectively, highly homologous to the respective antiporters of Vibrio species and E. coli. We showed that both Vc-NhaA and Vc-NhaB confer Na+ resistance and that Vc-NhaA displays an antiport activity in E. coli, which is similar in magnitude, kinetic parameters, and pH regulation to that of E. coli NhaA. To determine the roles of the Na+/H+ antiporters in V. cholerae, we constructed nhaA, nhaB, and nhaD mutants (single, double, and triple mutants). In contrast to E. coli, the inactivation of the three putative antiporter genes (Vc-nhaABD) in V. cholerae did not alter the bacterial exponential growth in the presence of high Na+ concentrations and had only a slight effect in the stationary phase. In contrast, a pronounced and similar Li+-sensitive phenotype was found with all mutants lacking Vc-nhaA during the exponential phase of growth and also with the triple mutant in the stationary phase of growth. By using 2-n-nonyl-4-hydroxyquinoline N-oxide, a specific inhibitor of the electron-transport-linked Na+ pump NADH-quinone oxidoreductase (NQR), we determined that in the absence of NQR activity, the Vc-NhaA Na+/H+ antiporter activity becomes essential for the resistance of V. cholerae to Na+ at alkaline pH. Since the ion pump NQR is Na+ specific, we suggest that its activity masks the Na+/H+ but not the Li+/H+ antiporter activities. Our results indicate that the Na+ resistance of the human pathogen V. cholerae requires a complex molecular system involving multiple antiporters and the NQR pump. PMID- 12562794 TI - Analysis of DNA binding and transcriptional activation by the LysR-type transcriptional regulator CbbR of Xanthobacter flavus. AB - The LysR-type transcriptional regulator CbbR controls the expression of the cbb and gap-pgk operons in Xanthobacter flavus, which encode the majority of the enzymes of the Calvin cycle required for autotrophic CO2 fixation. The cbb operon promoter of this chemoautotrophic bacterium contains three potential CbbR binding sites, two of which partially overlap. Site-directed mutagenesis and subsequent analysis of DNA binding by CbbR and cbb promoter activity were used to show that the potential CbbR binding sequences are functional. Inverted repeat IR1 is a high-affinity CbbR binding site. The main function of this repeat is to recruit CbbR to the cbb operon promoter. In addition, it is required for negative autoregulation of cbbR expression. IR3 represents the main low-affinity binding site of CbbR. Binding to IR3 occurs in a cooperative manner, since mutations preventing the binding of CbbR to IR1 also prevent binding to the low-affinity site. Although mutations in IR3 have a negative effect on the binding of CbbR to this site, they result in an increased promoter activity. This is most likely due to steric hindrance of RNA polymerase by CbbR since IR3 partially overlaps with the -35 region of the cbb operon promoter. Mutations in IR2 do not affect the DNA binding of CbbR in vitro but have a severe negative effect on the activity of the cbb operon promoter. This IR2 binding site is therefore critical for transcriptional activation by CbbR. PMID- 12562795 TI - Characterization of extradiol dioxygenases from a polychlorinated biphenyl degrading strain that possess higher specificities for chlorinated metabolites. AB - Recent studies demonstrated that 2,3-dihydroxybiphenyl 1,2-dioxygenase from Burkholderia sp. strain LB400 (DHBDLB400; EC 1.13.11.39) cleaves chlorinated 2,3 dihydroxybiphenyls (DHBs) less specifically than unchlorinated DHB and is competitively inhibited by 2',6'-dichloro-2,3-dihydroxybiphenyl (2',6'-diCl DHB). To determine whether these are general characteristics of DHBDs, we characterized DHBDP6-I and DHBDP6-III, two evolutionarily divergent isozymes from Rhodococcus globerulus strain P6, another good polychlorinated biphenyl (PCB) degrader. In contrast to DHBDLB400, both rhodococcal enzymes had higher specificities for some chlorinated DHBs in air-saturated buffer. Thus, DHBDP6-I cleaved the DHBs in the following order of specificity: 6-Cl DHB > 3'-Cl DHB approximately DHB approximately 4'-Cl DHB > 2'-Cl DHB > 4-Cl DHB > 5-Cl DHB. It also cleaved its preferred substrate, 6-Cl DHB, three times more specifically than DHB. Interestingly, some of the worst substrates for DHBDP6-I were among the best for DHBDP6-III (4-Cl DHB > 5-Cl DHB approximately 6-Cl DHB approximately 3'-Cl DHB > DHB > 2'-Cl DHB approximately 4'-Cl DHB; DHBDP6-III cleaved 4-Cl DHB two times more specifically than DHB). Generally, each of the monochlorinated DHBs inactivated the enzymes more rapidly than DHB. The exceptions were 4-Cl DHB for DHBDP6-I and 2'-Cl DHB for DHBDP6-III. As observed in DHBDLB400, chloro substituents influenced the reactivity of the dioxygenases with O2. For example, the apparent specificities of DHBDP6-I and DHBDP6-III for O2 in the presence of 2'-Cl DHB were lower than those in the presence of DHB by factors of >60 and 4, respectively. DHBDP6-I and DHBDP6-III shared the relative inability of DHBDLB400 to cleave 2',6'-diCl DHB (apparent catalytic constants of 0.088 +/- 0.004 and 0.069 +/- 0.002 s(-1), respectively). However, these isozymes had remarkably different apparent K(m) values for this compound (0.007 +/- 0.001, 0.14 +/- 0.01, and 3.9 +/- 0.4 micro M for DHBDLB400, DHBDP6-I, and DHBDP6-III, respectively). The markedly different reactivities of DHBDP6-I and DHBDP6-III with chlorinated DHBs undoubtedly contribute to the PCB-degrading activity of R. globerulus P6. PMID- 12562796 TI - Pyoverdine-mediated regulation of FpvA synthesis in Pseudomonas aeruginosa: involvement of a probable extracytoplasmic-function sigma factor, FpvI. AB - A search of the pvd pyoverdine biosynthesis locus of Pseudomonas aeruginosa identified an open reading frame, PA2387, whose product exhibited a sequence similar to those of a number of so-called extracytoplasmic- function sigma factors responsible for siderophore-dependent expression of iron-siderophore receptors in Escherichia coli and Pseudomonas putida. Deletion of this gene, dubbed fpvI, compromised pyoverdine-dependent FpvA ferric pyoverdine receptor production and fpvA gene expression, while the cloned gene stimulated fpvA expression. A Fur-binding site was identified immediately upstream of fpvI, consistent with the observed iron-regulated expression of fpvI and fpvA. PMID- 12562797 TI - Molecular characteristics of spontaneous deletions in the hyperthermophilic archaeon Sulfolobus acidocaldarius. AB - Prokaryotic genomes acquire and eliminate blocks of DNA sequence by lateral gene transfer and spontaneous deletion, respectively. The basic parameters of spontaneous deletion, which are expected to influence the course of genome evolution, have not been determined for any hyperthermophilic archaeon. We therefore screened a number of independent pyrimidine auxotrophs of Sulfolobus acidocaldarius for deletions and sequenced those detected. Deletions accounted for only 0.4% of spontaneous pyrE mutations, corresponding to a frequency of about 10(-8) per cell. Nucleotide sequence analysis of five independent deletions showed no significant association of the endpoints with short direct repeats, despite the fact that several such repeats occur within the pyrE gene and that duplication mutations in pyrE reverted at high frequencies. Endpoints of the spontaneous deletions did not coincide with short inverted repeats or potential stem-loop structures. No consensus sequence common to all the deletions could be identified, although two deletions showed the potential of being stabilized by octanucleotide sequences elsewhere in pyrE, and another pair of deletions shared an octanucleotide at their 3' ends. The unusually low frequency and low sequence dependence of spontaneous deletions in the S. acidocaldarius pyrE gene compared to other genetic systems could not be explained in terms of possible constraints imposed by the 5-fluoroorotate selection. PMID- 12562798 TI - Transcriptional switch on of ssgA by A-factor, which is essential for spore septum formation in Streptomyces griseus. AB - A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) triggers morphological development and secondary metabolism in Streptomyces griseus. A transcriptional activator (AdpA) in the A-factor regulatory cascade switches on a number of genes required for both processes. AdBS11 was identified in a library of the DNA fragments that are bound by AdpA and mapped upstream of ssgA, which is essential for septum formation in aerial hyphae. Gel mobility shift assays and DNase I footprinting revealed three AdpA-binding sites at nucleotide positions about -235 (site 1), -110 (site 2), and +60 (site 3) with respect to the transcriptional start point, p1, of ssgA. ssgA had two transcriptional start points, one starting at 124 nucleotides (p1) and the other starting at 79 nucleotides (p2) upstream of the start codon of ssgA. Of the three binding sites, only sites 1 and 2 were required for transcriptional activation of p1 and p2 by AdpA. The transcriptional switch on of ssgA required the extracytoplasmic function sigma factor, sigma(AdsA), in addition to AdpA. However, it was unlikely that sigma(AdsA) recognized the two ssgA promoters, since their -35 and -10 sequences were not similar to the promoter sequence motifs recognized by sigma(BldN), a sigma(AdsA) homologue of Streptomyces coelicolor A3(2). An ssgA disruptant formed aerial hyphae, but did not form spores, irrespective of the carbon source of the medium, which indicated that ssgA is a member of the whi genes. Transcriptional analysis of ssfR, located just upstream of ssgA and encoding an IclR-type transcriptional regulator, suggested that no read-through from ssfR into ssgA occurred, and ssgA was transcribed in the absence of ssfR. ssgA was thus found to be controlled by AdpA and not by SsfR to a detectable extent. SsfR appeared to regulate spore septum formation independently of SsgA or through interaction with SsgA in some unknown way, because an ssfR disruptant also showed a whi phenotype. PMID- 12562799 TI - The small subunit of M. AquI is responsible for sequence-specific DNA recognition and binding in the absence of the catalytic domain. AB - AquI DNA methyltransferase (M. AquI) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine to the C5 position of the outermost deoxycytidine base in the DNA sequence 5'-CCCGGG-3'. M. AquI is a heterodimer in which the polypeptide chain is separated at the junction between the two equivalent structural domains in the related enzyme M. HhaI. Recently, we reported the subcloning, overexpression, and purification of the subunits (alpha and beta) of M. AquI separately. Here we describe the DNA binding properties of M. AquI. The results presented here indicate that the beta subunit alone contains all of the information for sequence-specific DNA recognition and binding. The first step in the sequence-specific recognition of DNA by M. AquI involves the formation of binary complex with the target recognition domain in conjunction with conserved sequence motifs IX and X, found in all known C5 DNA methyltransferases, contained in the beta subunit. The alpha subunit enhances the binding of the beta subunit to DNA specifically and nonspecifically. It is likely that the addition of the alpha subunit to the beta subunit stabilizes the conformation of the beta subunit and thereby enhances its affinity for DNA indirectly. Addition of S-adenosyl-L methionine and its analogues S-adenosyl-L-homocysteine and sinefungin enhances binding, but only in the presence of the alpha subunit. These compounds did not have any effect on DNA binding by the beta subunit alone. Using a 30-mer oligodeoxynucleotide substrate containing 5-fluorodeoxycytidine (5-FdC), it was found that the beta subunit alone did not form a covalent complex with its specific sequence in the absence or presence of S-adenosyl-L-methionine. However, the addition of the alpha subunit to the beta subunit led to the formation of a covalent complex with specific DNA sequence containing 5-FdC. PMID- 12562800 TI - KtrAB and KtrCD: two K+ uptake systems in Bacillus subtilis and their role in adaptation to hypertonicity. AB - Recently, a new type of K+ transporter, Ktr, has been identified in the bacterium Vibrio alginolyticus (T. Nakamura, R. Yuda, T. Unemoto, and E. P. Bakker, J. Bacteriol. 180:3491-3494, 1998). The Ktr transport system consists of KtrB, an integral membrane subunit, and KtrA, a subunit peripherally bound to the cytoplasmic membrane. The genome sequence of Bacillus subtilis contains two genes for each of these subunits: yuaA (ktrA) and ykqB (ktrC) encode homologues to the V. alginolyticus KtrA protein, and yubG (ktrB) and ykrM (ktrD) encode homologues to the V. alginolyticus KtrB protein. We constructed gene disruption mutations in each of the four B. subtilis ktr genes and used this isogenic set of mutants for K+ uptake experiments. Preliminary K+ transport assays revealed that the KtrAB system has a moderate affinity with a Km value of approximately 1 mM for K+, while KtrCD has a low affinity with a Km value of approximately 10 mM for this ion. A strain defective in both KtrAB and KtrCD exhibited only a residual K+ uptake activity, demonstrating that KtrAB and KtrCD systems are the major K+ transporters of B. subtilis. Northern blot analyses revealed that ktrA and ktrB are cotranscribed as an operon, whereas ktrC and ktrD, which occupy different locations on the B. subtilis chromosome, are expressed as single transcriptional units. The amount of K+ in the environment or the salinity of the growth medium did not influence the amounts of the various ktr transcripts. A strain with a defect in KtrAB is unable to cope with a sudden osmotic upshock, and it exhibits a growth defect at elevated osmolalities which is particularly pronounced when KtrCD is also defective. In the ktrAB strain, the osmotically mediated growth defect was associated with a rapid loss of K+ ions from the cells. Under these conditions, the cells stopped synthesizing proteins but the transcription of the osmotically induced proHJ, opuA, and gsiB genes was not impaired, demonstrating that a high cytoplasmic K+ concentration is not essential for the transcriptional activation of these genes at high osmolarity. Taken together, our data suggest that K+ uptake via KtrAB and KtrCD is an important facet in the cellular defense of B. subtilis against both suddenly imposed and prolonged osmotic stress. PMID- 12562801 TI - Genetic and biochemical analysis of phosphatase activity of Escherichia coli NRII (NtrB) and its regulation by the PII signal transduction protein. AB - Mutant forms of Escherichia coli NRII (NtrB) were isolated that retained wild type NRII kinase activity but were defective in the PII-activated phosphatase activity of NRII. Mutant strains were selected as mimicking the phenotype of a strain (strain BK) that lacks both of the related PII and GlnK signal transduction proteins and thus has no mechanism for activation of the NRII phosphatase activity. The selection and screening procedure resulted in the isolation of numerous mutants that phenotypically resembled strain BK to various extents. Mutations mapped to the glnL (ntrB) gene encoding NRII and were obtained in all three domains of NRII. Two distinct regions of the C-terminal, ATP-binding domain were identified by clusters of mutations. One cluster, including the Y302N mutation, altered a lid that sits over the ATP-binding site of NRII. The other cluster, including the S227R mutation, defined a small surface on the "back" or opposite side of this domain. The S227R and Y302N proteins were purified, along with the A129T (NRII2302) protein, which has reduced phosphatase activity due to a mutation in the central domain of NRII, and the L16R protein, which has a mutation in the N-terminal domain of NRII. The S227R, Y302N, and L16R proteins were specifically defective in the PII-activated phosphatase activity of NRII. Wild-type NRII, Y302N, A129T, and L16R proteins bound to PII, while the S227R protein was defective in binding PII. This suggests that the PII-binding site maps to the "back" of the C-terminal domain and that mutation of the ATP-lid, central domain, and N-terminal domain altered functions necessary for the phosphatase activity after PII binding. PMID- 12562802 TI - Whole-genome sequence variation among multiple isolates of Pseudomonas aeruginosa. AB - Whole-genome shotgun sequencing was used to study the sequence variation of three Pseudomonas aeruginosa isolates, two from clonal infections of cystic fibrosis patients and one from an aquatic environment, relative to the genomic sequence of reference strain PAO1. The majority of the PAO1 genome is represented in these strains; however, at least three prominent islands of PAO1-specific sequence are apparent. Conversely, approximately 10% of the sequencing reads derived from each isolate fail to align with the PAO1 backbone. While average sequence variation among all strains is roughly 0.5%, regions of pronounced differences were evident in whole-genome scans of nucleotide diversity. We analyzed two such divergent loci, the pyoverdine and O-antigen biosynthesis regions, by complete resequencing. A thorough analysis of isolates collected over time from one of the cystic fibrosis patients revealed independent mutations resulting in the loss of O-antigen synthesis alternating with a mucoid phenotype. Overall, we conclude that most of the PAO1 genome represents a core P. aeruginosa backbone sequence while the strains addressed in this study possess additional genetic material that accounts for at least 10% of their genomes. Approximately half of these additional sequences are novel. PMID- 12562803 TI - Effects of the chromosome partitioning protein Spo0J (ParB) on oriC positioning and replication initiation in Bacillus subtilis. AB - Spo0J (ParB) of Bacillus subtilis is a DNA-binding protein that belongs to a conserved family of proteins required for efficient plasmid and chromosome partitioning in many bacterial species. We found that Spo0J contributes to the positioning of the chromosomal oriC region, but probably not by recruiting the origin regions to specific subcellular locations. In wild-type cells during exponential growth, duplicated origin regions were generally positioned around the cell quarters. In a spo0J null mutant, sister origin regions were often closer together, nearer to midcell. We found, by using a Spo0J-green fluorescent protein [GFP] fusion, that the subcellular location of Spo0J was a consequence of the chromosomal positions of the Spo0J binding sites. When an array of binding sites (parS sites) were inserted at various chromosomal locations in the absence of six of the eight known parS sites, Spo0J-GFP was no longer found predominantly at the cell quarters, indicating that Spo0J is not sufficient to recruit chromosomal parS sites to the cell quarters. spo0J also affected chromosome positioning during sporulation. A spo0J null mutant showed an increase in the number of cells with some origin-distal regions located in the forespore. In addition, a spo0J null mutation caused an increase in the number of foci per cell of LacI-GFP bound to arrays of lac operators inserted in various positions in the chromosome, including the origin region, an increase in the DNA-protein ratio, and an increase in origins per cell, as determined by flow cytometry. These results indicate that the spo0J mutant produced a significant proportion of cells with increased chromosome content, probably due to increased and asynchronous initiation of DNA replication. PMID- 12562804 TI - Growth phase-coupled alterations in cell structure and function of Escherichia coli. AB - Escherichia coli cultures can be fractionated into more than 20 cell populations, each having a different bouyant density and apparently representing a specific stage of cell differentiation from exponential growth to stationary phase (H. Makinoshima, A. Nishimura, and A. Ishihama, Mol. Microbiol. 43:269-279, 2002). The density increase was found to be impaired at an early step for a mutant E. coli with the disrupted rpoS gene, which encodes the RNA polymerase RpoS (sigma S) for stationary-phase gene transcription. This finding suggests that RpoS is need for the entire process of cell density increase. In the absence of RpoF sigma factor, the flagella are not formed as observed by electron microscopy, but the growth phase-coupled density increase takes place as in wild-type E. coli, confirming that the alteration in cell density is not directly correlated with the presence or absence of flagella. In the stationary-phase cells, accumulation of electron-dense areas was observed by electron microscopic observation of bacterial thin sections. By chemical determination, the increase in glycogen (or polysaccharides) was suggested to be one component, which contributes to the increase in weight-to-volume ratio of stationary-phase E. coli cells. PMID- 12562805 TI - Glycerol-3-phosphate acquisition in spirochetes: distribution and biological activity of glycerophosphodiester phosphodiesterase (GlpQ) among Borrelia species. AB - Relapsing-fever spirochetes achieve high cell densities (>10(8)/ml) in their host's blood, while Lyme disease spirochetes do not (<10(5)/ml). This striking contrast in pathogenicity of these two groups of bacteria suggests a fundamental difference in their ability to either exploit or survive in blood. Borrelia hermsii, a tick-borne relapsing-fever spirochete, contains orthologs to glpQ and glpT, genes that encode glycerophosphodiester phosphodiesterase (GlpQ) and glycerol-3-phosphate transporter (GlpT), respectively. In other bacteria, GlpQ hydrolyzes deacylated phospholipids to glycerol-3-phosphate (G3P) while GlpT transports G3P into the cytoplasm. Enzyme assays on 17 isolates of borreliae demonstrated GlpQ activity in relapsing-fever spirochetes but not in Lyme disease spirochetes. Southern blots demonstrated glpQ and glpT in all relapsing-fever spirochetes but not in the Lyme disease group. A Lyme disease spirochete, Borrelia burgdorferi, that was transformed with a shuttle vector containing glpTQ from B. hermsii produced active enzyme, which demonstrated the association of glpQ with the hydrolysis of phospholipids. Sequence analysis of B. hermsii identified glpF, glpK, and glpA, which encode the glycerol facilitator, glycerol kinase, and glycerol-3-phosphate dehydrogenase, respectively, all of which are present in B. burgdorferi. All spirochetes examined had gpsA, which encodes the enzyme that reduces dihydroxyacetone phosphate (DHAP) to G3P. Consequently, three pathways for the acquisition of G3P exist among borreliae: (i) hydrolysis of deacylated phospholipids, (ii) reduction of DHAP, and (iii) uptake and phosphorylation of glycerol. The unique ability of relapsing-fever spirochetes to hydrolyze phospholipids may contribute to their higher cell densities in blood than those of Lyme disease spirochetes. PMID- 12562806 TI - Detection of other microbial species by Salmonella: expression of the SdiA regulon. AB - Salmonella, Escherichia, and Klebsiella do not encode any recognized type of N acylhomoserine lactone (AHL) synthase, and consistent with this, they do not synthesize AHLs under any conditions tested. However, they do encode an AHL receptor of the LuxR family, named SdiA. MudJ fusions in four loci are known to respond to plasmid-encoded sdiA in Salmonella, but only the rck locus has been described. Here we report the location and sequence analysis of the remaining three loci. The srg-6::MudJ is within gtgA of the gifsy-2 prophage, and the srg 7::MudJ is within PSLT61 of the virulence plasmid. Both fusions are in the antisense orientation. The third fusion, srgE5::MudJ, is within a horizontally acquired gene of unknown function at 33.6 centisomes that we have named srgE. Previously, sdiA expressed from its natural position in the chromosome was demonstrated to activate a plasmid-based transcriptional fusion to the rck promoter in response to AHL production by other bacterial species. However, the MudJ fusions did not respond to chromosomal sdiA. Here we report that MudJ fusions to three of the four loci (not srg-6) are activated by AHL in an sdiA dependent manner during growth in motility agar (0.25% agar) but not during growth in top agar (0.7% agar) or on agar plates (1.2% agar). In motility agar, the srgE promoter responds to sdiA at 30 degrees C and higher while the rck and srg-7 promoters respond only at 37 or 42 degrees C. Substantial AHL-independent SdiA activity was observed at 30 degrees C but not at 37 degrees C. PMID- 12562807 TI - Modulation of pPS10 host range by plasmid-encoded RepA initiator protein. AB - We report here the isolation and analysis of novel repA host range mutants of pPS10, a plasmid originally found in Pseudomonas savastanoi. Upon hydroxylamine treatment, five plasmid mutants were selected for their establishment in Escherichia coli at 37 degrees C, a temperature at which the wild-type form cannot be established. The mutations were located in different functional regions of the plasmid RepA initiation protein, and the mutants differ in their stable maintenance, copy number, and ability to interact with sequences of the basic replicon. Four of them have broadened their host range, and one of them, unable to replicate in Pseudomonas, has therefore changed its host range. Moreover, the mutants also have increased their replication efficiency in strains other than E. coli such as Pseudomonas putida and Alcaligenes faecalis. None of these mutations drastically changed the structure or thermal stability of the wild-type RepA protein, but in all cases an enhanced interaction with host-encoded DnaA protein was detected by gel filtration chromatography. The effects of the mutations on the functionality of RepA protein are discussed in the framework of a three dimensional model of the protein. We propose possible explanations for the host range effect of the different repA mutants, including the enhancement of limiting interactions of RepA with specific host replication factors such as DnaA. PMID- 12562808 TI - Role of two novel two-component regulatory systems in development and phosphatase expression in Myxococcus xanthus. AB - We have cloned a two-component regulatory system (phoR2-phoP2) of Myxococcus xanthus while searching for genes that encode proteins with phosphatase activity, where phoR2 encodes the histidine kinase and phoP2 encodes the response regulator. A second system, phoR3-phoP3, was identified and isolated by using phoP2 as a probe. These two systems are quite similar, sharing identities along the full-length proteins of 52% on the histidine kinases and 64% on the response regulators. The predicted structures of both kinases suggest that they are anchored to the membrane, with the sensor domains being located in the periplasmic space and the kinase domains in the cytoplasm. The response regulators (PhoP2 and PhoP3) exhibit a helix-loop-helix motif typical of DNA binding proteins in the effector domains located in the C-terminal region. Studies on two single-deletion mutants and one double-deletion mutant have revealed that these systems are involved in development. Mutant fruiting bodies are not well packed, originating loose and flat aggregates where some myxospores do not reshape properly, and they remain as elongated cells. These systems are also involved in the expression of Mg-independent acid and neutral phosphatases, which are expressed during development. The neutral phosphatase gene is especially dependent on PhoP3. Neither PhoP2 nor PhoP3 regulates the expression of alkaline phosphatases and the pph1 gene. PMID- 12562809 TI - Identification and characterization of a Vibrio cholerae gene, mbaA, involved in maintenance of biofilm architecture. AB - The formation of biofilms is thought to play a key role in the environmental survival of the marine bacterium Vibrio cholerae. Although the factors involved in V. cholerae attachment to abiotic surfaces have been extensively studied, relatively little is known about the mechanisms involved in the subsequent maturation of the biofilms. Here we report the identification of a novel gene, which we have named mbaA (for maintenance of biofilm architecture), that plays a role in the formation and maintenance of the highly organized three-dimensional architecture of V. cholerae El Tor biofilms. We demonstrate that although the absence of mbaA does not significantly affect the initial attachment of cells onto the surface, it leads to the formation of biofilms that lack the typical structure, including the pillars of cells separated by fluid-filled channels that are evident in mature wild-type biofilms. Microscopic analysis indicates that the absence of mbaA leads to an increase in the amount of extracellular matrix material in the biofilms. The predicted mbaA product is a member of a family of regulatory proteins, containing GGDEF and EAL domains, suggesting that MbaA regulates the synthesis of some component of the biofilm matrix. PMID- 12562810 TI - Subcellular localization of a small sporulation protein in Bacillus subtilis. AB - SpoVM is an unusually small (26-residue-long) protein that is produced in the mother cell chamber of the sporangium during the process of sporulation in Bacillus subtilis. We investigated the subcellular localization of SpoVM, which is believed to be an amphipathic alpha-helix, by using a fusion of the sporulation protein to the green fluorescence protein (GFP). We found that SpoVM GFP is recruited to the polar septum shortly after the sporangium undergoes asymmetric division and that the fusion protein localizes to the mother cell membrane that surrounds the forespore during the subsequent process of engulfment. We identified a patch of three residues near the N terminus of the proposed alpha-helix that is needed both for proper subcellular localization and for SpoVM function. We also identified a patch of residues on the opposite face of the helix and residues near both ends of the protein that are needed for SpoVM function but not for subcellular localization. Subcellular localization of SpoVM GFP was found to require an unknown gene(s) under the control of the mother cell transcription factor sigmaE. We propose that the N-terminal patch binds to an unknown anchoring protein that is produced under the control of sigmaE and that other residues important in SpoVM function to recruit an unknown sporulation protein(s) to the mother cell membrane that surrounds the forespore. Our results provide evidence that SpoVM function depends on proper subcellular localization. PMID- 12562811 TI - Biofilm growth and detachment of Actinobacillus actinomycetemcomitans. AB - The gram-negative, oral bacterium Actinobacillus actinomycetemcomitans has been implicated as the causative agent of several forms of periodontal disease in humans. When cultured in broth, fresh clinical isolates of A. actinomycetemcomitans form tenacious biofilms on surfaces such as glass, plastic, and saliva-coated hydroxyapatite, a property that probably plays an important role in the ability of this bacterium to colonize the oral cavity and cause disease. We examined the morphology of A. actinomycetemcomitans biofilm colonies grown on glass slides and in polystyrene petri dishes by using light microscopy and scanning and transmission electron microscopy. We found that A. actinomycetemcomitans developed asymmetric, lobed biofilm colonies that displayed complex architectural features, including a layer of densely packed cells on the outside of the colony and nonaggregated cells and large, transparent cavities on the inside of the colony. Mature biofilm colonies released single cells or small clusters of cells into the medium. These released cells adhered to the surface of the culture vessel and formed new colonies, enabling the biofilm to spread. We isolated three transposon insertion mutants which produced biofilm colonies that lacked internal, nonaggregated cells and were unable to release cells into the medium. All three transposon insertions mapped to genes required for the synthesis of the O polysaccharide (O-PS) component of lipopolysaccharide. Plasmids carrying the complementary wild-type genes restored the ability of mutant strains to synthesize O-PS and release cells into the medium. Our findings suggest that A. actinomycetemcomitans biofilm growth and detachment are discrete processes and that biofilm cell detachment evidently involves the formation of nonaggregated cells inside the biofilm colony that are destined for release from the colony. PMID- 12562812 TI - cis Elements necessary for developmental expression of a Myxococcus xanthus gene that depends on C signaling. AB - Cell contact-mediated C signaling coordinates morphogenesis and gene expression during development of Myxococcus xanthus. One promoter that depends on C signaling for transcription lies upstream of Omega4403, the site of a Tn5 lac insertion in the genome. The Omega4403 promoter has a C-box sequence centered at 49 bp that matches the consensus 5'-CAYYCCY-3', which is found in several C signal-dependent promoters. Mutational analysis of the Omega4403 promoter region was performed to test the importance of the C box and to identify other cis acting elements. A 6-bp change in the -10 region eliminated promoter activity, but a 6-bp change in the -35 region decreased activity only about twofold. Certain single-base-pair changes in the C box centered at -49 bp abolished promoter activity, establishing the importance of this sequence element. Single base-pair changes in a C-box-like sequence centered at -77 bp also abolished promoter activity, but the pattern of mutational effects was different from that for the C box centered at -49 bp. Additional single-base-pair changes indicated that all 10 bp from -79 to -70 bp are important for Omega4403 promoter activity. Mutations at -59, -61, -62, and -63 bp also abolished promoter activity, defining a 5-bp element from -63 to -59 bp. This 5-bp element is separated from the 10-bp element (i.e., -79 to -70 bp) by 6 bp that can be changed without loss of promoter activity. Likewise, the 5 bp between the 5-bp element and the C box can be changed without loss of activity, but deletion of these 5 bp abolished activity, indicating that spacing is important. Sequences similar to the 5- and 10-bp elements, as well as the C box, are present in other C-signal-dependent promoters, suggesting some similarity in the regulatory mechanisms, but there are also indications that these cis elements do not function identically in the different promoters. PMID- 12562813 TI - ldpA encodes an iron-sulfur protein involved in light-dependent modulation of the circadian period in the cyanobacterium Synechococcus elongatus PCC 7942. AB - We generated random transposon insertion mutants to identify genes involved in light input pathways to the circadian clock of the cyanobacterium Synechococcus elongatus PCC 7942. Two mutants, AMC408-M1 and AMC408-M2, were isolated that responded to a 5-h dark pulse differently from the wild-type strain. The two mutants carried independent transposon insertions in an open reading frame here named ldpA (for light-dependent period). Although the mutants were isolated by a phase shift screening protocol, the actual defect is a conditional alteration in the circadian period. The mutants retain the wild-type ability to phase shift the circadian gene expression (bioluminescent reporter) rhythm if the timing of administration of the dark pulse is corrected for a 1-h shortening of the circadian period in the mutant. Further analysis indicated that the conditional short-period mutant phenotype results from insensitivity to light gradients that normally modulate the circadian period in S. elongatus, lengthening the period at low light intensities. The ldpA gene encodes a polypeptide that predicts a 7Fe-8S cluster-binding motif expected to be involved in redox reactions. We suggest that the LdpA protein modulates the circadian clock as an indirect function of light intensity by sensing changes in cellular physiology. PMID- 12562814 TI - Peptidoglycan synthesis in the absence of class A penicillin-binding proteins in Bacillus subtilis. AB - Penicillin-binding proteins (PBPs) catalyze the final, essential reactions of peptidoglycan synthesis. Three classes of PBPs catalyze either trans-, endo-, or carboxypeptidase activities on the peptidoglycan peptide side chains. Only the class A high-molecular-weight PBPs have clearly demonstrated glycosyltransferase activities that polymerize the glycan strands, and in some species these proteins have been shown to be essential. The Bacillus subtilis genome sequence contains four genes encoding class A PBPs and no other genes with similarity to their glycosyltransferase domain. A strain lacking all four class A PBPs has been constructed and produces a peptidoglycan wall with only small structural differences from that of the wild type. The growth rate of the quadruple mutant is much lower than those of strains lacking only three of the class A PBPs, and increases in cell length and frequencies of wall abnormalities were noticeable. The viability and wall production of the quadruple-mutant strain indicate that a novel enzyme can perform the glycosyltransferase activity required for peptidoglycan synthesis. This activity was demonstrated in vitro and shown to be sensitive to the glycosyltransferase inhibitor moenomycin. In contrast, the quadruple-mutant strain was resistant to moenomycin in vivo. Exposure of the wild type strain to moenomycin resulted in production of a phenotype similar to that of the quadruple mutant. PMID- 12562815 TI - Identification of genes required for synthesis of the adhesive holdfast in Caulobacter crescentus. AB - Adhesion to both abiotic and biotic surfaces by the gram-negative prothescate bacterium Caulobacter crescentus is mediated by a polar organelle called the "holdfast," which enables the bacterium to form stable monolayer biofilms. The holdfast, a complex polysaccharide composed in part of N-acetylglucosamine, localizes to the tip of the stalk (a thin cylindrical extension of the cell wall and membranes). We report here the isolation of adhesion mutants with transposon insertions in an uncharacterized gene cluster involved in holdfast biogenesis (hfs) as well as in previously identified polar development genes (podJ and pleC), and the holdfast attachment genes (hfa). Clean deletions of three of the four genes in the hfs gene cluster (hfsDAB) resulted in a severe holdfast biogenesis phenotype. These mutants do not bind to surfaces or to a fluorescently labeled lectin, specific for N-acetylglucosamine. Transmission electron microscopy indicated that the hfsDAB mutants fail to synthesize a holdfast at the stalk tip. The predicted hfs gene products have significant sequence similarity to proteins necessary for exopolysaccharide export in gram-negative bacteria. HfsA has sequence similarity to GumC from Xanthomonas campestris, which is involved in exopolysaccharide export in the periplasm. HfsD has sequence similarity to Wza from Escherichia coli, an outer membrane protein involved in secretion of polysaccharide through the outer membrane. HfsB is a novel protein involved in holdfast biogenesis. These data suggest that the hfs genes play an important role in holdfast export. PMID- 12562817 TI - Differentiation and anaerobiosis in standing liquid cultures of Streptomyces coelicolor. AB - Streptomyces coelicolor differentiates on solid agar media by forming aerial hyphae that septate into spores. We here show that differentiation also occurs in standing liquid minimal media. After a period of submerged growth, hyphae migrate to the air interface, where they become fixed by a rigid reflecting film. Colonies that result from these hyphae form sporulating aerial hyphae. In addition, submerged hyphae in the liquid minimal medium may attach to the surface. Liquid standing cultures easily become anoxic only 1 to 2 mm below the surface. Yet, biomass increases, implying the existence of metabolic pathways supporting anaerobic growth. PMID- 12562816 TI - Proteomic analysis of the spore coats of Bacillus subtilis and Bacillus anthracis. AB - The outermost proteinaceous layer of bacterial spores, called the coat, is critical for spore survival, germination, and, for pathogenic spores, disease. To identify novel spore coat proteins, we have carried out a preliminary proteomic analysis of Bacillus subtilis and Bacillus anthracis spores, using a combination of standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation and improved two-dimensional electrophoretic separations, followed by matrix assisted laser desorption ionization-time of flight and/or dual mass spectrometry. We identified 38 B. subtilis spore proteins, 12 of which are known coat proteins. We propose that, of the novel proteins, YtaA, YvdP, and YnzH are bona fide coat proteins, and we have renamed them CotI, CotQ, and CotU, respectively. In addition, we initiated a study of coat proteins in B. anthracis and identified 11 spore proteins, 6 of which are candidate coat or exosporium proteins. We also queried the unfinished B. anthracis genome for potential coat proteins. Our analysis suggests that the B. subtilis and B. anthracis coats have roughly similar numbers of proteins and that a core group of coat protein species is shared between these organisms, including the major morphogenetic proteins. Nonetheless, a significant number of coat proteins are probably unique to each species. These results should accelerate efforts to develop B. anthracis detection methods and understand the ecological role of the coat. PMID- 12562818 TI - 2-aminopurine allows interspecies recombination by a reversible inactivation of the Escherichia coli mismatch repair system. AB - 2-Aminopurine treatment of Escherichia coli induces a reversible phenotype of DNA mismatch repair deficiency. This transient phenotype results in a 300-fold increase in the frequency of interspecies conjugational recombination with a Salmonella enterica serovar Typhimurium Hfr donor. This method can be used for the generation of biodiversity by allowing recombination between diverged genes and genomes. PMID- 12562819 TI - Identification and characterization of the gerH operon of Bacillus anthracis endospores: a differential role for purine nucleosides in germination. AB - We identified a tri-cistronic operon, gerH, in Bacillus anthracis that is important for endospore germination triggered by two distinct germination response pathways termed inosine-His and purine-Ala. Together, the two pathways allow B. anthracis endospores a broader recognition of purines and amino acids that may be important for host-mediated germination. PMID- 12562820 TI - The periplasmic protein MppA requires an additional mutated locus to repress marA expression in Escherichia coli. AB - Escherichia coli strain TP985, which has an insertional mutation in the gene for the periplasmic murein tripeptide binding protein MppA, was previously reported to overproduce MarA and exhibit a multiple-antibiotic resistance (Mar) phenotype (H. Li and J. T. Park, J. Bacteriol. 181:4842-4847, 1999). We found that TP985 contained a previously unrecognized marR mutation which was responsible for the Mar phenotype. Transduction of the mppA mutation from TP985 to another wild-type strain did not affect antibiotic susceptibility. Overproduction of MppA repressed marA transcription in TP985 but not in other mppA or marR mutants. Therefore, TP985 contains an additional unknown mutation(s) that facilitates the repression of marA expression by MppA. PMID- 12562821 TI - Overrepresentation of a gene family encoding extracytoplasmic solute receptors in Bordetella. AB - A family of genes that are likely to encode extracytoplasmic solute receptors is strongly overrepresented in several beta-proteobacteria, including Bordetella pertussis. This gene family, of which members have been called bug genes, contains some examples that are contained within polycistronic operons coding for tripartite uptake transporters of the TTT family, while the vast majority are "orphan" genes. Proteomic and functional analyses demonstrated that several of these genes are expressed in B. pertussis, and one is involved in citrate uptake. The bug genes probably form an ancient family that has been subjected to a large expansion in a restricted phylogenic group. PMID- 12562822 TI - Corrected sequence of the bacteriophage p22 genome. AB - We report the first accurate genome sequence for bacteriophage P22, correcting a 0.14% error rate in previously determined sequences. DNA sequencing technology is now good enough that genomes of important model systems like P22 can be sequenced with essentially 100% accuracy with minimal investment of time and resources. PMID- 12562824 TI - Comparison of the intestinal uptake of cholesterol, plant sterols, and stanols in mice. AB - The recent identification of the aberrant transport proteins ABCG5 and ABCG8 resulting in sitosterolemia suggests that intestinal uptake of cholesterol is an unselective process, and that discrimination between cholesterol and plant sterols takes place at the level of sterol efflux from the enterocyte. Although plant sterols are structurally very similar to cholesterol, differing only in their side chain length, they are absorbed from the intestine to a markedly lower extent. In order to further evaluate the process of discrimination, three different sterols (cholesterol, campesterol, sitosterol) and their corresponding 5 alpha-stanols (cholestanol, campestanol, sitostanol) were compared concerning their concentration in the proximal small intestine, in serum, and in bile after a single oral dose of deuterated compounds. The data obtained support the hypothesis that i) the uptake of sterols and stanols is an extremely rapid process, ii) discrimination probably takes place on the level of reverse transport back into the gut lumen, iii) plant stanols are taken up, but not absorbed to a measurable extent, and iv) the process of discrimination probably also exists at the level of biliary excretion. The range of structural alterations that decrease intestinal absorption and increase biliary excretion is: 1) campesterol, 2) cholestanol-sitosterol, and 3) campestanol-sitostanol. PMID- 12562823 TI - Prokaryotic utilization of the twin-arginine translocation pathway: a genomic survey. AB - The twin-arginine translocation (Tat) pathway, which has been identified in plant chloroplasts and prokaryotes, allows for the secretion of folded proteins. However, the extent to which this pathway is used among the prokaryotes is not known. By using a genomic approach, a comprehensive list of putative Tat substrates for 84 diverse prokaryotes was established. Strikingly, the results indicate that the Tat pathway is utilized to highly varying extents. Furthermore, while many prokaryotes use this pathway predominantly for the secretion of redox proteins, analyses of the predicted substrates suggest that certain bacteria and archaea secrete mainly nonredox proteins via the Tat pathway. While no correlation was observed between the number of Tat machinery components encoded by an organism and the number of predicted Tat substrates, it was noted that the composition of this machinery was specific to phylogenetic taxa. PMID- 12562825 TI - Appearance of atypical 3 alpha,6 beta,7 beta,12 alpha-tetrahydroxy-5 beta-cholan 24-oic acid in spgp knockout mice. AB - Bile formation and its canalicular secretion are essential functions of the mammalian liver. The sister-of-p-glycoprotein (spgp) gene was shown to encode the canalicular bile salt export protein, and mutations in spgp gene were identified as the cause of progressive familial intrahepatic cholestasis type 2. However, target inactivation of spgp gene in mice results in nonprogressive but persistent cholestasis and causes the secretion of unexpectedly large amounts of unknown tetrahydroxylated bile acid in the bile. The present study confirms the identity of this tetrahydroxylated bile acid as 3 alpha,6 beta,7 beta,12 alpha tetrahydroxy-5 beta-cholan-24-oic acid. The data further show that in serum, liver, and urine of the spgp knockout mice, there is a significant increase in the concentration of total bile salts containing a large amount of tetrahydroxy-5 beta-cholan-24-oic acid. The increase in total bile acids was associated with up regulation of the mRNA of cholesterol 7 alpha-hydroxylase in male mice only. It is suggested that the lower severity of the cholestasis in the spgp knockout mice may be due to the synthesis of 3 alpha,6 beta,7 beta,12 alpha-tetrahydroxy-5 beta cholan-24-oic acid, which neutralizes in part the toxic effect of bile acids accumulated in the liver. PMID- 12562826 TI - Different modulation of phospholipase A2 activity by saturated and monounsaturated N-acylethanolamines. AB - The physiological functions of N-acylethanolamines (NAEs) are poorly understood, although many functions were suggested for these naturally occurring membrane components of plants and animals. The binding with cannabinoid receptors CB1 and CB2 was demonstrated for some NAEs, such as anandamide. However, the chemical nature of these molecules suggests that some of their biological effects on biomembranes could be related, at least partially, to physical interactions with the lipid bilayer. The present work studies the effect of saturated and monounsaturated NAEs on phospholipase A2 (PLA2) activity, which is dependent on lipid bilayer features. The present study, performed by 2-dimethylamino-(6 lauroyl)-naphthalene (Laurdan) fluorescence, demonstrates that the acyl chain length and the presence of a single double bond are crucial for the enzymatic activity modulation by NAEs. In fact, saturated NAEs with 10 carbon atoms don't affect the PLA2 activity, while NAEs with 12 and 16 carbon atoms largely activate the enzyme. On the other hand, an acyl chain length of 18 carbon atoms, with or without the presence of a double bond, only slightly affects the enzymatic activity. A structural model for NAE-lipid interactions is proposed in order to explain the differences in PLA2 activity modulation by these fatty acid derivatives. PMID- 12562827 TI - Red blood cell fatty acid ethyl esters: a significant component of fatty acid ethyl esters in the blood. AB - Although alcohol abuse is known to cause an array of ethanol-induced red blood cell (RBC) abnormalities, the underlying molecular mechanisms remain poorly understood. Fatty acid ethyl esters (FAEEs) are toxic, nonoxidative ethanol metabolites that have been found in blood, plasma, and tissues. Because FAEEs have been shown to be incorporated into phospholipid bilayers, we conducted a controlled ethanol intake study to test the hypothesis that FAEEs accumulate and persist within RBCs following ethanol ingestion. We demonstrated that RBC FAEEs account for approximately 5% to 20% of total whole-blood FAEEs, and that the fatty acid composition of FAEEs in RBCs and plasma are different and vary differently over time. These data indicate that a significant percentage of FAEEs in the blood is associated with RBCs and that the metabolism of RBC FAEEs and that of plasma FAEEs (bound to albumin or lipoproteins) are largely independent. PMID- 12562828 TI - A cluster of eight hydroxysteroid dehydrogenase genes belonging to the aldo-keto reductase supergene family on mouse chromosome 13. AB - A subclass of hydroxysteroid dehydrogenases (HSD) are NADP(H)-dependent oxidoreductases that belong to the aldo-keto reductase (AKR) superfamily. They are involved in prereceptor or intracrine steroid modulation, and also act as bile acid-binding proteins. The HSD family members characterized thus far in human and rat have a high degree of protein sequence similarity but exhibit distinct substrate specificity. Here we report the identification of nine murine AKR genes in a cluster on chromosome 13 by a combination of molecular cloning and in silico analysis of this region. These include four previously isolated mouse HSD genes (Akr1c18, Akr1c6, Akr1c12, Akr1c13), the more distantly related Akr1e1, and four novel HSD genes. These genes exhibit highly conserved exon/intron organization and protein sequence predictions indicate 75% amino acid similarity. The previously identified AKR protein active site residues are invariant among all nine proteins, but differences are observed in regions that have been implicated in determining substrate specificity. Differences also occur in tissue expression patterns, with expression of some genes restricted to specific tissues and others expressed at high levels in multiple tissues. Our findings dramatically expand the repertoire of AKR genes and identify unrecognized family members with potential roles in the regulation of steroid metabolism. PMID- 12562829 TI - Plasma ceramide and lysophosphatidylcholine inversely correlate with mortality in sepsis patients. AB - Recent data indicate that ceramide (Cer) and lysophosphatidylcholine (LPC) regulate immune cell functions. Since these bioactive lipids are generated in blood plasma by inflammatory lipases, we hypothesized that they may be involved in the process of acute systemic sepsis. In order to provide support for this hypothesis, we analyzed the plasma levels of Cer and LPC by quantitative tandem mass spectrometry in 102 sepsis patients starting with the day at which the sepsis criteria were fulfilled for the first time, as well as on day 4 and day 11. The values were compared with 56 healthy controls and correlated with sepsis related mortality within 30 days of study entry. Most Cer species were increased in sepsis patients, while all LPC species were markedly decreased. In addition, we determined the molar ratios with their precursor molecules sphingomyelin (SPM) and phosphatidylcholine (PC), which reflect the enzymatic reactions responsible for their formation. Species-specific as well as total Cer-SPM ratios were increased, whereas LPC-PC ratios were decreased in sepsis patients. The increased Cer-SPM ratios as well as the decreased LPC-PC ratios showed a strong predictive power for sepsis-related mortality. Together with existing data from in vitro experiments and animal models, the results provide the first ex vivo indication for the role of Cer and lysophospholipids in systemic inflammation in humans. PMID- 12562830 TI - Dietary fat modulation of apoA-II metabolism and prevention of senile amyloidosis in the senescence- accelerated mouse. AB - Senescence-accelerated mouse-prone (SAMP1; SAMP1@Umz) is an animal model of senile amyloidosis with apolipoprotein A-II (apoA-II) amyloid fibril (AApoAII) deposits. This study was undertaken to investigate the effects of dietary fats on AApoAII deposits in SAMP1 mice when purified diets containing 4% fat as butter, safflower oil, or fish oil were fed to male mice for 26 weeks. The serum HDL cholesterol was significantly lower (P < 0.01) in mice on the diet containing fish oil (7.4 +/- 3.0 mg/dl) than in mice on the butter diet (38.7 +/- 12.5 mg/dl), which in turn had significantly lower (P < 0.01) HDL levels than mice on the safflower oil diet (51.9 +/- 5.6 mg/dl). ApoA-II was also significantly lower (P < 0.01) in mice on the fish oil diet (7.6 +/- 2.7 mg/dl) than on the butter (26.9 +/- 7.3 mg/dl) or safflower oil (21.6 +/- 3.7 mg/dl) diets. The mice fed fish oil had a significantly greater ratio (P < 0.01) of apoA-I to apoA-II, and a smaller HDL particle size than those fed butter and safflower oil. Severe AApoAII deposits in the spleen, heart, skin, liver, and stomach were shown in the fish oil group compared with those in the butter and safflower oil groups (fish oil > butter > safflower oil group, P < 0.05). These findings suggest that dietary fats differ in their effects on serum lipoprotein metabolism, and that dietary lipids may modulate amyloid deposition in SAMP1 mice. PMID- 12562831 TI - Prooxidant and antioxidant properties of human serum ultrafiltrates toward LDL: important role of uric acid. AB - Oxidized LDL is present within atherosclerotic lesions, demonstrating a failure of antioxidant protection. A normal human serum ultrafiltrate of Mr below 500 was prepared as a model for the low Mr components of interstitial fluid, and its effects on LDL oxidation were investigated. The ultrafiltrate (0.3%, v/v) was a potent antioxidant for native LDL, but was a strong prooxidant for mildly oxidized LDL when copper, but not a water-soluble azo initiator, was used to oxidize LDL. Adding a lipid hydroperoxide to native LDL induced the antioxidant to prooxidant switch of the ultrafiltrate. Uric acid was identified, using uricase and add-back experiments, as both the major antioxidant and prooxidant within the ultrafiltrate for LDL. The ultrafiltrate or uric acid rapidly reduced Cu2+ to Cu+. The reduction of Cu2+ to Cu+ may help to explain both the antioxidant and prooxidant effects observed. The decreased concentration of Cu2+ would inhibit tocopherol-mediated peroxidation in native LDL, and the generation of Cu+ would promote the rapid breakdown of lipid hydroperoxides in mildly oxidized LDL into lipid radicals. The net effect of the low Mr serum components would therefore depend on the preexisting levels of lipid hydroperoxides in LDL. These findings may help to explain why LDL oxidation occurs in atherosclerotic lesions in the presence of compounds that are usually considered to be antioxidants. PMID- 12562832 TI - Immunochemical evidence that human apoB differs when expressed in rodent versus human cells. AB - LDL from human apolipoprotein B-100 (apoB-100) transgenic (HuBTg+/+) mice contains more triglyceride than LDL from normolipidemic subjects. To obtain novel monoclonal antibody (MAb) probes of apoB conformation, we generated hybridomas from HuBTg+/+ that had been immunized with LDL isolated from human plasma. One apoE-specific and four anti-apoB-100-specific hybridomas were identified. Two MAbs, 2E1 and 3D11, recognized an epitope in the amino-terminal 689 residues of apoB in native apoB-containing lipoproteins (LpBs) from human plasma or from the supernatant of human hepatoma HepG2 cells, but did not react with LpB from HuBTg+/+ mice or LpB secreted by human apoB-100-transfected rat McArdle 7777 hepatoma cells. 2E1 reacted weakly and 3D11 reacted strongly with apoB from HuBTg+/+ mice after SDS-PAGE. The lack of expression of the 2E1 and 3D11 epitopes on native LpB from HuBTg+/+ mice did not solely reflect the abnormal lipid composition of murine LpB. Both epitopes were detected in all human plasma samples tested and in all human plasma LpB classes. Therefore, human apoB expressed by rodent hepatocytes or hepatoma cells appears to adopt a different conformation or undergoes different posttranslational modification than apoB expressed in human hepatocytes or hepatoma cells. PMID- 12562833 TI - Postprandial recruitment of neutrophils may contribute to endothelial dysfunction. AB - Atherosclerosis is a low-grade inflammatory disease involving leukocytes, lipids, and glucose leading to endothelial dysfunction. Since activation of neutrophils by triglycerides and glucose has been described in vitro, we hypothesized that the postprandial phase is an inflammatory state affecting leukocytes, possibly contributing to endothelial dysfunction. We measured postprandial blood leukocyte counts, cytokines, hydroperoxides (HPOs), and flow-mediated vasodilation (FMD) in eight healthy males (age 23 +/- 2 years) after a FAT (50 g/m2) and GLUCOSE challenge (37.5 g/m2), a combination of both (MIXED test), and after WATER. All tests, except WATER, resulted in significantly impaired FMD (10% reduction) between t = 1 h and t = 3 h, accompanied by a significant increase of neutrophils (59% after FAT and 28% after GLUCOSE and MIXED), total plasma HPOs (15 to 31% increase), and plasma interleukin-8 (IL-8) (50-130% increase). WATER did not affect FMD, neutrophils, HPOs, or IL-8. Lymphocytes increased gradually in all tests (40-70% increase at t = 10 h compared with t = 0; P < 0.005), paralleling a gradual 3- to 5-fold interleukin-6 increase. Monocyte and erythrocyte counts did not change in any test. In conclusion, the neutrophil increment during postprandial lipemia and glycemia with concomitant IL-8 and HPO increases may contribute to endothelial dysfunction. Lymphocyte increment is a nonspecific diurnal process. Postprandial intravascular inflammatory changes may be relevant for the pathogenesis of atherosclerosis. PMID- 12562834 TI - Apolipoprotein composition and particle size affect HDL degradation by chymase: effect on cellular cholesterol efflux. AB - Mast cell chymase, a chymotrypsin-like neutral protease, can proteolyze HDL3. Here we studied the ability of rat and human chymase to proteolyze discoidal pre beta-migrating reconstituted HDL particles (rHDLs) containing either apolipoprotein A-I (apoA-I) or apoA-II. Both chymases cleaved apoA-I in rHDL at identical sites, either at the N-terminus (Tyr18 or Phe33) or at the C-terminus (Phe225), so generating three major truncated polypeptides that remained bound to the rHDL. The cleavage sites were independent of the size of the rHDL particles, but small particles were more susceptible to degradation than bigger ones. Chymase-induced truncation of apoA-I yielded functionally compromised rHDL with reduced ability to promote cellular cholesterol efflux. In sharp contrast to apoA I, apoA-II was resistant to degradation. However, when apoA-II was present in rHDL that also contained apoA-I, it was degraded by chymase. We conclude that chymase reduces the ability of apoA-I in discoidal rHDL particles to induce cholesterol efflux by cleaving off either its amino- or carboxy-terminal portion. This observation supports the concept that limited extracellular proteolysis of apoA-I is one pathophysiologic mechanism leading to the generation and maintenance of foam cells in atherosclerotic lesions. PMID- 12562835 TI - Separate myocardial ethanolamine phosphotransferase activities responsible for plasmenylethanolamine and phosphatidylethanolamine synthesis. AB - Ethanolamine phosphotransferase (EPT) is a key enzyme responsible for the synthesis of ethanolamine glycerophospholipids. Plasmenylethanolamine is a predominant molecular subclass of ethanolamine glycerophospholipids in the heart. The present study was designed to identify the selective use of 1-O-alk-1'-enyl-2 acyl-sn-glycerol as a substrate for EPT as a mechanism responsible for the predominance of plasmenylethanolamine in the rabbit heart. EPT activity in rabbit myocardial membranes using 1,2-diacyl-sn-glycerol as substrate is activated by Mn2+, inhibited by dithiobisnitrobenzoic acid (DTNB) and is unaffected by Ca2+. In contrast, ethanolamine phosphotransferase activity using 1-O-alk-1'-enyl-2 acyl-sn-glycerol as substrate is inhibited by Mn2+ and Ca2+, but is activated by DTNB. Additionally, ethanolamine phosphotransferase activity using 1-O-alk-1' enyl-2-acyl-sn-glycerol substrate was more sensitive to thermal denaturation compared with that of 1,2-diacyl-sn-glycerol. Taken together, these results suggest that separate ethanolamine phosphotransferase activities are present in heart membranes that are responsible for the synthesis of phosphatidylethanolamine and plasmenylethanolamine. PMID- 12562836 TI - Effects of LDL enriched with different dietary fatty acids on cholesteryl ester accumulation and turnover in THP-1 macrophages. AB - LDL enriched with either saturated, monounsaturated, n-6 polyunsaturated, or n-3 polyunsaturated fatty acids were used to study the effects of dietary fatty acids on macrophage cholesteryl ester (CE) accumulation, physical state, hydrolysis, and cholesterol efflux. Incubation of THP-1 macrophages with acetylated LDL (AcLDL) from each of the four diet groups resulted in both CE and triglyceride (TG) accumulation, in addition to alterations of cellular CE, TG, and phospholipid fatty acyl compositions reflective of the individual LDLs. Incubation with monounsaturated LDL resulted in significantly higher total and CE accumulation when compared with the other groups. After TG depletion, intracellular anisotropic lipid droplets were visible in all four groups, with 71% of the cells incubated with monounsaturated AcLDL containing anisotropic lipid droplets, compared with 30% of cells incubated with n-3 AcLDL. These physical state differences translated into higher rates of both CE hydrolysis and cholesterol efflux in the n-3 group. These data suggest that monounsaturated fatty acids may enhance atherosclerosis by increasing both cholesterol delivery to macrophage foam cells and the percentage of anisotropic lipid droplets, while n-3 PUFAs decrease atherosclerosis by creating more fluid cellular CE droplets that accelerate the rate of CE hydrolysis and the efflux of cholesterol from the cell. PMID- 12562837 TI - Serum paraoxonase: effect of the apolipoprotein composition of HDL and the acute phase response. AB - Genetic variations of paraoxonase (PON) correlate with HDL cholesterol and apolipoprotein A-I (apoA-I), suggesting antiatherogenic properties. Atherosclerosis occurs naturally in humans and rabbits but not in mice. We compared variations of PON arylesterase activity (PON AEase, phenylacetate substrate) in humans, rabbits, and mice. In humans and rabbits, >95% of PON AEase is HDL associated. In mice, about 30% of PON AEase is lipid poor. In the absence of apoA-I in mice, total PON AEase is reduced and >60% is lipid poor. PON AEase level and distribution is restored in apoA-I-/- mice injected with adenoviruses encoding human apoA-I and in transgenic mice expressing human apoA-I at a steady state level. Thus, while apoA-I is not required for the HDL association of PON AEase, induced variations in apoA-I correlate with changes in HDL-associated, but not lipid-poor, PON AEase. PON AEase associates only with apoA-I- or apoE containing HDL but not VLDL. In the absence of both apoA-I and apoE, PON AEase is all-lipid-poor. PON AEase is displaced from HDL by ultracentrifugation and following incubation with serum amyloid A. Variations in the PON distribution between HDL and lipid-poor fractions may have important consequences in its antioxidant activity and in atherogenesis. PMID- 12562838 TI - Side chain oxidized oxysterols in cerebrospinal fluid and the integrity of blood brain and blood-cerebrospinal fluid barriers. AB - The side chain oxidized oxysterol 24S-hydroxycholesterol (24-OH-chol) is formed almost exclusively in the brain, and there is a continuous passage of this oxysterol through the circulation to the liver. 27-Hydroxycholesterol (27-OH chol) is produced in most organs and is also taken up by the liver. The 27-OH chol-24-OH-chol ratio is about 0.1 in the brain and about 2 in the circulation. This ratio was found to be about 0.4 in cerebrospinal fluid (CSF) of asymptomatic patients, consistent with a major contribution from the circulation in the case of 27-OH-chol. In accordance with this, we demonstrated a significant flux of deuterium labeled 27-OH-chol from plasma to the CSF in a healthy volunteer. Patients with a defective blood-brain barrier were found to have markedly increased absolute levels (up to 10-fold) of both 27-OH-chol and 24-OH-chol in CSF, with a ratio between the two sterols reaching up to 2. There was a significant positive correlation between the levels of both oxysterols in CSF and the albuminCSF-albuminplasma ratio. The 27-OH-cholCSF-24-OH-cholCSF ratio was found to be about normal in patients with active multiple sclerosis and significantly increased in patients with meningitis, polyneuropathy, or hemorrhages. Results are discussed in relation to the possible use of 24-OH cholCSF as a surrogate marker of central nervous system demyelination and/or neuronal death. PMID- 12562839 TI - Single session exercise stimulates formation of pre beta 1-HDL in leg muscle. AB - Physical activity can raise the level of circulating HDL cholesterol. Pre beta 1 HDL is thought to be either the initial acceptor of cellular cholesterol or virtually the first particle in the pathway of the formation of HDL from apolipoprotein A-I and cellular lipids. We have therefore sought to identify pre beta 1-HDL in arterial and venous circulations of exercising legs in healthy individuals and in subjects with stable Type 2 diabetes mellitus. Blood samples were taken simultaneously from the femoral artery and vein before and after 25 min cycling exercise. The major findings were, first, that exercise significantly increased plasma concentration of pre beta 1-HDL (20% increase, P < 0.05) and second, that the pre beta 1-HDL concentration was significantly higher in the venous compared with the arterial blood both before and after exercise in both diabetics and controls. In the combined population, formation of pre beta 1-HDL at rest was 9.9 +/- 5.2 mg/min and exercise enhanced pre beta 1-HDL formation 6.6 fold in both groups. PMID- 12562841 TI - Lack of a direct role for macrosialin in oxidized LDL metabolism. AB - Murine macrosialin (MS), a scavenger receptor family member, is a heavily glycosylated transmembrane protein expressed predominantly in macrophage late endosomes. MS is also found on the cell surface where it is suggested, on the basis of ligand blotting, to bind oxidized LDL (oxLDL). Here we report on the regulation of MS by an atherogenic high-fat diet and oxLDL, and on the inability of MS in transfected cells to bind oxLDL. MS expression was markedly increased in the livers of atherosclerosis-susceptible C57BL/6 and atherosclerosis-resistant C3H/HeJ mice fed an atherogenic high-fat diet. In resident-mouse peritoneal macrophages, treatment with oxLDL upregulated MS mRNA and protein expression 1.5- to 3-fold. MS, overexpressed in COS-7 cells through adenovirus mediated gene transfer, bound oxLDL by ligand blotting. However, no binding of oxLDL to MS was observed in intact transfected COS-7 and Chinese hamster ovary cells, despite significant cell surface expression of MS. Furthermore, inhibition of MS through gene silencing did not affect the binding of oxLDL to macrophages. We conclude that although MS expression in macrophages and Kupffer cells is responsive to a proatherogenic inflammatory diet and to oxLDL, MS does not function as an oxLDL receptor on the cell surface. PMID- 12562840 TI - Impact of simvastatin, niacin, and/or antioxidants on cholesterol metabolism in CAD patients with low HDL. AB - The HDL Atherosclerosis Treatment Study (HATS) demonstrated a clinical benefit in coronary artery disease patients with low HDL cholesterol (HDL-C) levels treated with simvastatin and niacin (S-N) or S-N plus antioxidants (S-N+A) compared with antioxidants alone or placebo. Angiographically documented stenosis regressed in the S-N group but progressed in all other groups. To assess the mechanism(s) responsible for these observations, surrogate markers of cholesterol absorption and synthesis were measured in a subset of 123 HATS participants at 24 months (on treatment) and at 38 months (off treatment). Treatment with S-N reduced desmosterol and lathosterol levels (cholesterol synthesis indicators) 46% and 36% (P < 0.05), respectively, and elevated campesterol and beta-sitosterol levels (cholesterol absorption indicators) 70% and 59% (P < 0.05), respectively, relative to placebo and antioxidant but not S-N+A. Treatment with antioxidants alone had no significant effect. Combining S-N with antioxidants reduced desmosterol and lathosterol by 37% and 31%, and elevated campesterol and beta sitosterol levels by 54% and 46%, but differences did not attain significance. Mean change in percent stenosis was positively associated with a percent change in lathosterol (r = 0.26, P < 0.005) and negatively associated with a percent change in beta-sitosterol (r = -0.21, P < 0.01). These data suggest that changes in stenosis were attributable, in part, to changes in cholesterol metabolism. PMID- 12562842 TI - Caveolin-1 does not affect SR-BI-mediated cholesterol efflux or selective uptake of cholesteryl ester in two cell lines. AB - Free cholesterol (FC) has been reported to efflux from cells through caveolae, which are 50-100 nm plasma membrane pits. The 22 kDa protein caveolin-1 is concentrated in caveolae and is required for their formation. The HDL scavenger receptor BI (SR-BI), which stimulates both FC efflux and selective uptake of HDL derived cholesteryl ester (CE), has been reported to be concentrated in caveolae, suggesting that this localization facilitates flux of FC and CE across the membrane. However, we found that overexpression of caveolin-1 in Fischer rat thyroid (FRT) cells, which lack caveolin-1 and caveolae, or HEK 293 cells, which normally express very low levels of caveolin-1, did not affect FC efflux to HDL or liposomes. Transient expression of SR-B1 did not affect this result. Similarly, caveolin-1 expression did not affect selective uptake of CE from labeled HDL particles in FRT or HEK 293 cells transfected with SR-BI. We conclude that basal and SR-BI-stimulated FC efflux to HDL and liposomes and SR-BI-mediated selective uptake of HDL CE are not affected by caveolin-1 expression in HEK 293 or FRT cells. PMID- 12562843 TI - Apolipoprotein[a] secretion from hepatoma cells is regulated in a size-dependent manner by alterations in disulfide bond formation. AB - Apolipoprotein[a] (apo[a]) is a large disulfide linked glycoprotein synthesized by hepatocytes. We have examined the role of disulfide bond formation in the processing of apo[a] using human and rat hepatoma cells expressing apo[a] isoforms containing varying numbers of kringle 4 (K4) domains, following treatment with DTT. Hepatoma cells expressing 6- or 9-K4 isoforms revealed approximately 90% inhibition of apo[a] secretion following DTT treatment, although larger isoforms containing 13- or 17-K4 domains demonstrated continued secretion (up to 30% of control values), suggesting that a fraction of the larger isoforms is at least partially DTT resistant. Wash-out experiments demonstrated that these effects were completely reversible for all isoforms studied, with no enhanced degradation associated with prolonged intracellular retention. DTT treatment was associated with enhanced binding of apo[a] with the endoplasmic reticulum-associated chaperone proteins calnexin, calreticulin, and BiP, which was reversible upon DTT removal. The chemical chaperone 6-aminohexanoic acid, previously demonstrated by others to rescue defective apo[a] secretion associated with alterations in glycosylation, failed to alter the secretion of apo[a] following DTT treatment. The demonstration that DTT modulates apo[a] secretion in a manner influenced by both the type and number of K4 repeats extends understanding of the mechanisms that regulate its exit from the endoplasmic reticulum. PMID- 12562844 TI - Differences in the regulation of adipose tissue and liver lipogenesis by carbohydrates in humans. AB - We assessed the contributions of human liver and adipose tissue de novo lipogenesis (DNL) to triacylglycerol (TAG) synthesis. Volunteers were fed a high energy, high-carbohydrate diet (HC, n = 5) or a normocaloric diet (NC, n = 10). NC subjects remained in the fasting state (Study 1, n = 5) or received oral glucose (Study 2, n = 5) throughout the test (12 h). HC subjects remained in the fasting state (Study 3). They ingested deuterated water and [U-13C]acetate to trace lipogenesis. Adipose tissue fatty-acid (FA) synthase (FAS), acetyl-CoA carboxylase 1 (ACC1), and SREBP-1c mRNA were measured. Plasma TAG-FA was labeled by 13C and deuterium showing active liver lipogenesis, which was stimulated (P < 0.05) by oral glucose and HC diet. Adipose tissue TAG had no detectable 13C enrichment in any test, showing no significant incorporation of TAG-FA provided by liver lipogenesis, but were labeled by deuterium in all tests, showing active DNL in situ; however, rough quantitative estimates showed that adipose DNL was minimal (<1 g), and poorly stimulated by oral glucose or HC diet. mRNA levels were not increased by the HC diet. Adipose DNL is active in humans, but contributes little to TAG stores and is less responsive than liver DNL to stimulation by carbohydrates. PMID- 12562845 TI - Synthetic amphipathic helical peptides promote lipid efflux from cells by an ABCA1-dependent and an ABCA1-independent pathway. AB - In order to examine the necessary structural features for a protein to promote lipid efflux by the ABCA1 transporter, synthetic peptides were tested on ABCA1 transfected cells (ABCA1 cells) and on control cells. L-37pA, an l amino acid peptide that contains two class-A amphipathic helices linked by proline, showed a 4-fold increase in cholesterol and phospholipid efflux from ABCA1 cells compared to control cells. The same peptide synthesized with a mixture of l and d amino acids was less effective than L-37pA in solubilizing dimyristoyl phosphatidyl choline vesicles and in effluxing lipids. In contrast, the 37pA peptide synthesized with all d amino acids (D-37pA) was as effective as L-37pA. Unlike apoA-I, L-37pA and D-37pA were also capable, although at a reduced rate, of causing lipid efflux independent of ABCA1 from control cells, Tangier disease cells, and paraformaldehyde fixed ABCA1 cells. The ability of peptides to bind to cells correlated with their lipid affinity. In summary, the amphipathic helix was found to be a key structural motif for peptide-mediated lipid efflux from ABCA1, but there was no stereoselective requirement. In addition, unlike apoA-I, synthetic peptides can also efflux lipid by a passive, energy-independent pathway that does not involve ABCA1 but does depend upon their lipid affinity. PMID- 12562846 TI - Effect of fluid mechanical stresses and plasma constituents on aggregation of LDL. AB - LDL aggregates when exposed to even moderate fluid mechanical stresses in the laboratory, yet its half-life in the circulation is 2-3 days, implying that little aggregation occurs. LDL may be protected from aggregation in vivo by components of plasma, or by a qualitative difference in flows. Previous studies have shown that HDL and albumin inhibit the aggregation induced by vortexing. Using a more reproducible method of inducing aggregation and assessing aggregation both spectrophotometrically and by sedimentation techniques, we showed that at physiological concentrations, albumin is the more effective inhibitor, and that aggregation is substantially but not completely inhibited in plasma. Heat denatured and fatty-acid-stripped albumin were more effective inhibitors than normal albumin, supporting the idea that hydrophobic interactions are involved. Aggregation of LDL in a model reproducing several aspects of flow in the circulation was 200-fold slower, but was still inhibited by HDL and albumin, suggesting similar mechanisms are involved. Within the sensitivity of our technique, LDL aggregation did not occur in plasma exposed to these flows. Thus, as a result of the characteristics of blood flow and the inhibitory effects of plasma components, particularly albumin, LDL aggregation is unlikely to occur within the circulation. PMID- 12562847 TI - Inhibition of both the apical sodium-dependent bile acid transporter and HMG-CoA reductase markedly enhances the clearance of LDL apoB. AB - Discovery of the ileal apical sodium-dependent bile acid transporter (ASBT) permitted development of specific inhibitors of bile acid reabsorption, potentially a new class of cholesterol-lowering agents. In the present study, we tested the hypothesis that combining the novel ASBT inhibitor, SC-435, with the HMG-CoA reductase inhibitor, atorvastatin, would potentiate reductions in LDL cholesterol (LDL-C) and LDL apolipoprotein B (apoB). ApoB kinetic studies were performed in miniature pigs fed a typical human diet and treated with the combination of SC-435 (5 mg/kg/day) plus atorvastatin (3 mg/kg/day) (SC-435+A) or a placebo. SC-435+A decreased plasma total cholesterol by 23% and LDL-C by 40%. Multicompartmental analysis (SAAM II) demonstrated that LDL apoB significantly decreased by 35% due primarily to a 45% increase in the LDL apoB fractional catabolic rate (FCR). SC-435+A significantly decreased hepatic concentrations of free cholesterol and cholesteryl ester, and increased hepatic LDL receptor mRNA consequent to increased cholesterol 7alpha-hydroxylase expression and activity. In comparison, SC-435 (10 mg/kg/day) monotherapy decreased LDL apoB by 10% due entirely to an 18% increase in LDL apoB FCR, whereas atorvastatin monotherapy (3 mg/kg/day) decreased LDL apoB by 30% due primarily to a 22% reduction in LDL apoB production. We conclude that SC-435+A potentiates the reduction of LDL-C and LDL apoB due to complementary mechanisms of action. PMID- 12562848 TI - New perspectives on the regulation of intermembrane glycerophospholipid traffic. AB - In eukaryotes, phosphatidylserine (PtdSer) can serve as a precursor of phosphatidylethanolamine (PtdEtn) and phosphatidylcholine (PtdCho), which are the major cellular phospholipids. PtdSer synthesis originates in the endoplasmic reticulum (ER) and its subdomain named the mitochondria-associated membrane (MAM). PtdSer is transported to the mitochondria in mammalian cells and yeast, and decarboxylated by PtdSer decarboxylase 1 (Psd1p) to form PtdEtn. A second decarboxylase, Psd2p, is also found in yeast in the Golgi-vacuole. PtdEtn produced by Psd1p and Psd2p can be transported to the ER, where it is methylated to form PtdCho. Organelle-specific metabolism of the aminoglycerophospholipids is a powerful tool for experimentally following lipid traffic that is now enabling identification of new proteins involved in the regulation of this process. Genetic and biochemical experiments demonstrate that transport of PtdSer between the MAM and mitochondria is regulated by protein ubiquitination, which affects events at both membranes. Similar analyses of PtdSer transport to the locus of Psd2p now indicate that a membrane-bound phosphatidylinositol transfer protein and the C2 domain of Psd2p are both required on the acceptor membrane for efficient transport of PtdSer. Collectively, these recent findings indicate that novel multiprotein assemblies on both donor and acceptor membranes participate in interorganelle phospholipid transport. PMID- 12562849 TI - Lipid rafts: bringing order to chaos. AB - Lipid rafts are subdomains of the plasma membrane that contain high concentrations of cholesterol and glycosphingolipids. They exist as distinct liquid-ordered regions of the membrane that are resistant to extraction with nonionic detergents. Rafts appear to be small in size, but may constitute a relatively large fraction of the plasma membrane. While rafts have a distinctive protein and lipid composition, all rafts do not appear to be identical in terms of either the proteins or the lipids that they contain. A variety of proteins, especially those involved in cell signaling, have been shown to partition into lipid rafts. As a result, lipid rafts are thought to be involved in the regulation of signal transduction. Experimental evidence suggests that there are probably several different mechanisms through which rafts control cell signaling. For example, rafts may contain incomplete signaling pathways that are activated when a receptor or other required molecule is recruited into the raft. Rafts may also be important in limiting signaling, either by physical sequestration of signaling components to block nonspecific interactions, or by suppressing the intrinsic activity of signaling proteins present within rafts. This review provides an overview of the physical characteristics of lipid rafts and summarizes studies that have helped to elucidate the role of lipid rafts in signaling via receptor tyrosine kinases and G protein-coupled receptors. PMID- 12562850 TI - Lipid compositional analysis of pulmonary surfactant monolayers and monolayer associated reservoirs. AB - Pulmonary surfactant is a lipid:protein complex containing dipalmitoyl phosphatidylcholine (DPPC) as the major component. Recent studies indicate adsorbed surfactant films consist of a surface monolayer and a monolayer associated reservoir. It has been hypothesized that the monolayer and its functionally contiguous reservoir may be enriched in DPPC relative to bulk phase surfactant. We investigated the compositional relationship between the monolayer and its reservoir using paper-supported wet bridges to transfer films from adsorbing dishes to clean surfaces on spreading dishes. Spreading films appear to form monolayers in the spreading dishes. We employed bovine lipid extract surfactant [BLES(chol)] containing [3H]DPPC and either [14C]palmitoyl, oleoyl phosphatidylcholine (POPC), [14C]dipalmitoyl-phosphatidylglycerol (DPPG), [14C]palmitoyl, oleoyl-phosphatidylglycerol (POPG), or [14C]cholesterol. Radiolabeled phosphatidylglycerols were prepared using phospholipase D. The studies demonstrated that the [3H]DPPC-[14C] POPC ratios were the same in the prepared BLES dispersions as in Langmuir-Blodgett films, indicating a lack of DPPC selectivity during film formation. Furthermore, identical 3H-14C isotopic ratios were observed with DPPC and either 14C-labeled POPC, DPPG, POPG, or cholesterol in the original dispersions, the bulk phases in adsorption dish D1, and monolayers recovered from spreading dish D2. These relationships remained unperturbed with 2-fold increases in bulk concentrations in D1 and 10-fold variations in D1-D2 surface area. These results indicate adsorbed surfactant monolayers and their associated reservoirs possess similar lipid compositions and argue against selective adsorption of DPPC. PMID- 12562851 TI - Conversion of hexadecanoic acid to hexadecenoic acid by rat Delta 6-desaturase. AB - A higher content of C16:1 n-10 has recently been reported in the preputial gland of mice with a targeted disruption of the gene encoding stearoyl-CoA desaturase 1 (SCD1-/- mice) when compared with wild-type mice. This result has provided the first physiological evidence for the presence and regulation of a palmitoyl-CoA Delta 6-desaturase in mammals. To investigate the putative involvement of the known Delta 6-desaturase (FADS2) in this process, COS-7 cells expressing rat Delta 6-desaturase were incubated with C16:0. Transfected cells were able to synthesize C16:1 n-10, while nontransfected cells did not produce any C16:1 n-10. Evidence is therefore presented that the rat Delta 6-desaturase, which acts on the 18- and 24-carbon fatty acids of the n-6 and n-3 series, is also able to catalyze palmitic acid Delta 6 -desaturation. PMID- 12562852 TI - Lipid droplet targeting domains of adipophilin. AB - Adipophilin (ADPH), a prominent protein component of lipid storage droplets (LSDs), is postulated to be necessary for the formation and cellular function of these structures. The presence of significant sequence similarities within an approximately 100 amino acid region of the N-terminal portions of ADPH and related LSD binding proteins, perilipin and TIP47, has implicated this region, known as the "PAT" domain, in LSD targeting. Here we investigate the role of the PAT domain in targeting ADPH to LSDs by expressing this region, as well as selected N- and C-terminal truncations of mouse ADPH in COS7 cells as epitope tagged fusion proteins. Our studies show that truncations lacking either the PAT domain or the C-terminal half of ADPH both correctly targeted LSDs and increased the LSD content of transfected cells. Neither the PAT domain nor the C-terminal half of ADPH appeared to target LSDs or affect the LSD number. Instead, targeting fragments encompassed a putative alpha-helical region between amino acids 189 and 205, implicating this region in both LSD targeting and regulation of LSD formation. PMID- 12562853 TI - Analytical performance of a sandwich enzyme immunoassay for pre beta 1-HDL in stabilized plasma. AB - We have established an immunoassay for pre beta 1-HDL (the initial acceptor of cellular cholesterol) using a monoclonal antibody, MAb55201. Because pre beta 1 HDL is unstable during storage, fresh plasma must be used for pre beta 1-HDL measurements. In this study, we describe a method of stabilizing pre beta 1-HDL, and evaluate the analytical performance of the immunoassay for pre beta 1-HDL. Fresh plasma was stored under various conditions with or without a pretreatment consisting of a 21-fold dilution into 50% (v/v) sucrose. Pre beta 1-HDL concentration was measured by immunoassay. In nonpretreated samples, pre beta 1 HDL decreased significantly from the baseline after 6 h at room temperature. Although pre beta 1-HDL was more stable at 0 degrees C than at room temperature, it increased from 30.2 +/- 8.5 (SE) to 56.5 +/- 5.5 mg/l apolipoprotein A-I (apoA I) (P < 0.001) in hyperlipidemics, and from 18.4 +/- 1.2 to 37.9 +/- 3.3 mg/l apoA-I (P < 0.001) in normolipidemics after 5-day storage. After 30-day storage at -80 degrees C, pre beta 1-HDL increased from 29.0 +/- 4.0 to 38.0 +/- 5.7 mg/l apoA-I (P < 0.001) in hyperlipidemics, whereas it did not change in normolipidemics. In pretreated samples, pre beta 1-HDL concentration did not change significantly under any of the above conditions. Moreover, pre beta 1-HDL concentrations determined by immunoassay correlated with those determined by native two-dimensional gel electrophoresis (n = 24, r = 0.833, P < 0.05). An immunoassay using MAb55201 with pretreated plasma is useful for clinical measurement of pre beta 1-HDL. PMID- 12562854 TI - Novel mass spectrometric immunoassays for the rapid structural characterization of plasma apolipoproteins. AB - Novel mass spectrometric immunoassays (MSIAs) for the isolation and structural characterization of plasma apolipoprotein A-I (apoA-I), apoA-II, and apoE have been developed. The assays combine selective isolation of apolipoprotein species via affinity capture with mass-specific detection using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In application, plasma (from 50 microl of whole blood drawn from individuals, using finger lancet) was addressed with affinity pipette tips derivatized with antibodies toward the specific apolipoprotein. The time required for each assay was approximately 15 min, less if assays on multiple individuals were performed in parallel. In a brief study of five individuals, several recently reported apoA-II variants were identified and observed consistently in all individuals. Additionally, the apoE phenotype of E3/E3 was observed in three of the individuals, and E2/E3 and E3/E4 observed in the remaining two individuals, the latter of whom suffers from Alzheimer's disease. Overall, the MSIA approach offers a rapid, sensitive, and highly accurate means of profiling apolipoproteins from small volumes of plasma. PMID- 12562855 TI - Biotinylated theta-toxin derivative as a probe to examine intracellular cholesterol-rich domains in normal and Niemann-Pick type C1 cells. AB - BCtheta is a proteolytically nicked and biotinylated derivative of a cholesterol binding protein perfringolysin O (theta-toxin), and has been used to detect cholesterol-rich domains at the plasma membrane (PM). Here we show that by modifying the cell fixation condition, BCtheta can also be used to detect cholesterol-rich domains intracellularly. When cells were processed for PM cholesterol staining, the difference in BCtheta signals between the CT43 (CT) cell, a mutant Chinese hamster ovary cell line lacking the Niemann-Pick type C1 (NPC1) protein, and its parental cell 25RA (RA) was minimal. However, when cells were fixed with 4% paraformaldehyde, they became permeable to BCtheta. Under this condition, BCtheta mainly stained cholesterol-rich domains inside the cells, with the signal being much stronger in CT cells than in RA cells. The sensitivity of BCtheta staining was superior to that of filipin staining. The staining of cholesterol-rich domain(s) inside RA cells was sensitive to beta-cyclodextrin treatment, while most of the staining inside CT cells was relatively resistant to cyclodextrin treatment. Clear differences in intracellular BCtheta staining were also seen between the normal and mutant NPC1 fibroblasts of human or mouse origin. Thus, BCtheta is a powerful tool for visually monitoring cholesterol-rich domains inside normal and NPC cells. PMID- 12562856 TI - A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl CoA hydratase deficiency. AB - D-bifunctional protein (D-BP) plays an indispensable role in peroxisomal beta oxidation, and its inherited deficiency in humans is associated with severe clinical abnormalities. Three different subtypes of D-BP deficiency can be distinguished: 1) a complete deficiency of D-BP (type I), 2) an isolated D-BP enoyl-CoA hydratase deficiency (type II), and 3) an isolated D-BP 3-hydroxyacyl CoA dehydrogenase deficiency (type III). In this study, we developed a method to measure D-BP dehydrogenase activity independent of D-BP hydratase (D-BP HY) activity to distinguish between D-BP deficiency type I and type II, which until now was only possible by mutation analysis. For this assay, the hydratase domain of D-BP was expressed in the yeast Saccharomyces cerevisiae. After a coincubation of yeast homogenate expressing D-BP HY with fibroblast homogenate of patients using the enoyl-CoA ester of the bile acid intermediate trihydroxycholestanoic acid as substrate, D-BP dehydrogenase activity was measured. Fibroblasts of patients with a D-BP deficiency type II displayed D-BP dehydrogenase activity, whereas type I and type III patients did not. This newly developed assay to measure D-BP dehydrogenase activity in fibroblast homogenates provides a quick and reliable method to assign patients with deficient D-BP HY activity to the D BP deficiency subgroups type I or type II. PMID- 12562857 TI - Quantification of phosphatidic acid and lysophosphatidic acid by HPLC with evaporative light-scattering detection. AB - Phosphatidic acid (PA) and lysophosphatidic acid (LPA) are lipids that regulate cellular processes. PA stimulates kinases and may play a role in exocytosis and membrane fusion. LPA can induce cell proliferation, platelet aggregation, and microfilament formation. Due to the growing interest in these lipids, rapid purification and quantification of these lipids is desirable. We now describe a method that utilizes one HPLC run to separate trace amounts of PA and LPA from large amounts of lipids found in cellular extracts. A two-pump HPLC with a solvent system consisting of chloroform, methanol, water, and ammonium hydroxide was employed to produce a reliable, efficient purification of the two lipids. Lipid mass was quantified by a sensitive evaporative light-scattering detector. Using this new method, insulin addition increased both PA (87%) and LPA (217%) mass in Xenopus laevis oocytes. PMID- 12562858 TI - Monitoring hepatic cholesterol 7alpha-hydroxylase activity by assay of the stable bile acid intermediate 7alpha-hydroxy-4-cholesten-3-one in peripheral blood. AB - We describe an accurate method for monitoring the enzymatic activity of hepatic cholesterol 7alpha-hydroxylase (C7alphaOH; CYP7A1), the rate-limiting and major regulatory enzyme in the synthesis of bile acids. Assay of 7alpha-hydroxy-4 cholesten-3-one (C4), an intermediate in bile acid synthesis, revealed that the level of C4 in peripheral blood serum or plasma showed a strong correlation to the enzymatic activity of hepatic C7alphaOH, both at steady-state conditions (r = 0.929) as well as during the rapid changes that occur during the diurnal phases. This assay should be of value in clarifying the regulation of bile acid synthesis in vivo in laboratory animals and humans since it allows for the monitoring of hepatic C7alphaOH activity using peripheral blood samples. PMID- 12562859 TI - Different effects of n-6 and n-3 polyunsaturated fatty acids on the activation of rat smooth muscle cells by interleukin-1 beta. AB - There is good evidence that the n-3 polyunsaturated fatty acids (PUFAs) in fish oil have antiinflammatory effects and reduce the pathogenesis of atherosclerosis. However, the mechanisms underlying these actions are largely unknown. This study was designed to investigate the effects of membrane incorporation of two major components of fish oil [eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)], on rat smooth muscle cells (SMCs) activation induced by interleukin-1 beta (IL1 beta). We compared their effects with those of n-6 arachidonic acid (AA). Expression of vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1 adhesion molecules involved in SMCs migration was enhanced by AA, whereas EPA and DHA had no similar effects. We established that AA potentiates IL1 beta-induced expression of the type IIA secreted phospholipase A2 (sPLA2) gene, whereas EPA and DHA reduce this stimulation. EPA and DHA also abolished proinflammatory prostaglandin PGE2 production by inhibiting the IL1 beta-induced production of cyclooxygenase-2 (COX-2) mRNA. Much interest was then focused on three transcriptional factors implicated in inflammation control and especially in modulating rat sPLA2 and COX-2 gene transcription: nuclear factor kappa B, CCAAT/enhancer binding protein beta, and E26 transformation-specific-1. electrophoretic mobility shift assay revealed that the binding activity of all three factors was increased by AA and reduced (or not affected) by n-3 PUFA. These results indicate that EPA and DHA act in opposition to AA by modulating various steps of the inflammatory process induced by IL1 beta, probably by reducing mitogen-activated protein kinase p42/p44 activity. PMID- 12562860 TI - Effects on apoB-100 secretion and bile acid synthesis by redirecting cholesterol efflux from HepG2 cells. AB - This study determined the effects of apoA-I, HDL3, or hydroxy-beta-cyclodextrin on apoB-100 secretion and bile acid synthesis by HepG2 cells. The principal observations were that: 1) ApoB-100 secretion into the medium was significantly less after the addition of any of the three agents. 2) Triglyceride mass was not significantly changed from control in the medium but was significantly, although modestly, reduced in the cells. 3) Neither free cholesterol (FC) nor cholesteryl ester (CE) mass in the medium was changed; by contrast, CE mass was reduced within the cells although FC was not. 4) Although the total mass of cholesterol in the medium was unaffected, the proportion associated with apoB-100 was reduced, whereas the proportion associated with the non-apoB-100 fraction was increased. 5) There was also an unanticipated, but substantial, increase in bile acid synthesis induced by apoA-I, HDL3, or hydroxy-beta-cyclodextrin, which was time and concentration dependent, and which was associated with marked increases in cholesterol 7 alpha-hydroxylase activity. There were no significant changes in ACAT activity and only modest increases in HMG-CoA reductase activity. These findings support previous clinical observations that an elevated apoB-100 can accompany a low HDL cholesterol in normotriglyceridemic subjects. They also point to physiologically important, although still only partially understood, metabolic relationships amongst hepatic apoB-100 secretion, cholesterol efflux, and bile acid synthesis. PMID- 12562861 TI - Regulation of human delta-6 desaturase gene transcription: identification of a functional direct repeat-1 element. AB - The rate-limiting step in 20:4(n-6) and 22:6(n-3) synthesis is the desaturation of 18:2(n-6) and 18:3(n-3) by Delta-6 desaturase. In this report, we demonstrate that n-6 and n-3 PUFAs suppressed the hepatic expression of rodent Delta-6 desaturase by inhibiting the rate of Delta-6 desaturase gene transcription. In contrast, consumption of the peroxisome proliferator-activated receptor (PPAR)alpha activator WY 14,643 significantly enhanced the transcription of hepatic Delta-6 desaturase by more than 500%. Transfection reporter assays with HepG2 cells revealed that the PUFA response region for the human Delta-6 desaturase gene involved the proximal promoter region of -283/+1 human Delta-6 desaturase gene, while the WY 14,643 response element (RE) was identified as an imperfect direct repeat (DR-1) located at -385/-373. The WY 14,643 induction of the human Delta-6 desaturase promoter activity was dependent upon the expression of PPARalpha. Electrophoretic mobility shift assays revealed that nuclear proteins extracted from HepG2 cells expressing PPARalpha specifically interacted with the -385/-373 DR-1 sequence of the human Delta-6 desaturase gene. The interaction was eliminated by the unlabeled PPARalpha RE of the rat acyl-CoA oxidase gene, and the protein-DNA complex was super-shifted by treatment with anti-PPARalpha. The -385/-373 sequence also interacted with a mixture of in vitro translated PPARalpha-retinoic acid receptor X (RXR)alpha, but by themselves neither PPARalpha nor RXRalpha could bind to the Delta-6 desaturase DR-1. These data indicate that the 5'-flanking region of the human Delta-6 desaturase gene contains a DR-1 that functions in the regulation of human Delta-6 desaturase gene transcription, and thereby plays a role in the synthesis of 20- and 22-carbon polyenoic fatty acids. PMID- 12562862 TI - Linoleic acid supplementation of Barth syndrome fibroblasts restores cardiolipin levels: implications for treatment. AB - The object of this study was to investigate whether the levels of cardiolipin in cultured skin fibroblasts of patients with Barth syndrome (BTHS) can be restored by addition of linoleic acid to growth media. To this end, fibroblasts from controls and BTHS patients were grown in the presence or absence of linoleic acid. High-performance liquid chromatography-electrospray ionization tandem mass spectrometry was used for quantitative and compositional analysis of cardiolipin. Incubation of cells from both BTHS and controls with different concentrations of linoleic acid led to a dose- and time-dependent increase of cardiolipin levels. The increased levels of cardiolipin in fibroblasts of BTHS patients after treatment with linoleic acid indicate that an increased amount of linoleic acid in the diet might be beneficial to BTHS patients. PMID- 12562863 TI - Enhanced expression of hepatic lipogenic enzymes in an animal model of sedentariness. AB - The hindlimb-suspended rat was used as animal model to investigate the effects induced by immobilization of the skeletal muscle in the expression of the genes encoding hepatic lipogenic enzymes. Following a 14-day period of immobilization, rats were injected intraperitoneally with radioactive acetate, and the labeling of hepatic lipids and cholesterol was evaluated 15 min after the isotope injection. The incorporation of labeled acetate in lipids and cholesterol was almost three times higher in the liver of immobilized rats than in control animals as a consequence of the enhanced transcription of the genes encoding acetyl-CoA synthase, acetyl-CoA carboxylase, fatty acid synthase, and 3-hydroxy-3 methylglutaryl-CoA reductase. The high expression of the key enzymes for fatty acid and cholesterol synthesis induced by immobilization was not paralleled by an increase of the hepatic sterol-regulatory element binding protein (SREBP)-1 and SREBP-2 mRNA content. However, the expression of the mature form of SREBP-1 and SREBP-2 was higher in the nuclear fraction of immobilized rat liver than in controls due to a significant increase of the cleavage of the native proteins. Immobilization also affected the expression of proteins involved in lipid degradation. In fact, the hepatic content of peroxisome proliferator-activated receptor-alpha (PPARalpha) mRNA and of PPARalpha target genes encoding carnitine palmitoyl transferase-1 and acyl-CoA oxidase were significantly increased upon immobilization. PMID- 12562864 TI - Oxidized cholesterol in the diet is a source of oxidized lipoproteins in human serum. AB - The aim of this study was to determine in humans whether oxidized cholesterol in the diet is absorbed and contributes to the pool of oxidized lipids in circulating lipoproteins. When a meal containing 400 mg cholestan-5alpha,6alpha epoxy-3beta-ol (alpha-epoxy cholesterol) was fed to six controls and three subjects with Type III hyperlipoproteinemia, alpha-epoxy cholesterol in serum was found in chylomicron/chylomicron remnants (CM/RM) and endogenous (VLDL, LDL, and HDL) lipoproteins. In controls, alpha-epoxy cholesterol in CM/RM was decreased by 10 h, whereas in endogenous lipoproteins it remained in the circulation for 72 h. In subjects with Type III hyperlipoproteinemia, alpha-epoxy cholesterol was mainly in CM/RM. In vitro incubation of the CM/RM fraction containing alpha-epoxy cholesterol with human LDL and HDL that did not contain alpha-epoxy cholesterol resulted in a rapid transfer of oxidized cholesterol from CM/RM to both LDL and HDL. In contrast, no transfer was observed when human serum was substituted with rat serum, suggesting that cholesteryl ester transfer protein is mediating the transfer. Thus, alpha-epoxy cholesterol in the diet is incorporated into the CM/RM fraction and then transferred to LDL and HDL, contributing to lipoprotein oxidation. Moreover, LDL containing alpha-epoxy cholesterol displayed increased susceptibility to further copper oxidation in vitro. It is possible that oxidized cholesterol in the diet accelerates atherosclerosis by increasing oxidized cholesterol levels in circulating LDL and chylomicron remnants. PMID- 12562865 TI - Omega-3 fatty acid supplementation accelerates chylomicron triglyceride clearance. AB - Omega-3 fatty acids (FAs) reduce postprandial triacylglycerol (TG) concentrations. This study was undertaken to determine whether this effect was due to reduced production or increased clearance of chylomicrons. Healthy subjects (n = 33) began with a 4-week, olive oil placebo (4 g/d) run-in period. After a 4-week wash-out period, subjects were randomized to supplementation with 4 g/d of ethyl esters of either safflower oil (SAF), eicosapentaenoic acid (EPA), or docosahexaenoic acid (DHA) for 4 weeks. Results for EPA and DHA were similar, and therefore the data were combined into one omega-3 FA group. Omega-3 FA supplementation reduced the postprandial TG and apolipoprotein B (apo B)-48 and apoB-100 concentrations by 16% (P = 0.08), 28% (P < 0.001), and 24% (P < 0.01), respectively. Chylomicron TG half-lives in the fed state were reduced after omega 3 FA treatment (6.0 +/- 0.5 vs. 5.1 +/- 0.4 min; P < 0.05), but not after SAF (6.9 +/- 0.7 vs. 7.1 +/- 0.7 min). Omega-3 FA supplementation decreased chylomicron particle sizes (mean diameter; 293 +/- 44 vs. 175 +/- 25 nm; P < 0.01) and increased preheparin lipoprotein lipase (LPL; 0.6 +/- 0.1 vs. 0.9 +/- 0.1 micromol/h/ml; P < 0.05) activity during the fed state, but had no effect on postheparin LPL or hepatic lipase activities. The results suggest that omega-3 FA supplementation accelerates chylomicron TG clearance by increasing LPL activity, and that EPA and DHA are equally effective. PMID- 12562866 TI - Alkylacylglycerolipid domain of GPI molecules of Leishmania is responsible for inhibition of PKC-mediated c-fos expression. AB - Glycosylphosphatidylinositols (GPIs) are the most abundant molecules present in the membranes of the parasitic protozoa Leishmania responsible for multiple forms of leishmaniasis. Among the prominent biological activities displayed by the major Leishmania GPIs [lipophosphoglycan (LPG) and glycoinositolphospholipids (GIPLs)] is the inhibition of macrophage functions such as the protein kinase C (PKC)-dependent signaling pathway. The bioactivity of Leishmania GPIs is in contrast to Trypanosoma brucei and Plasmodium falciparum GPIs, which activate the macrophage functions. To address the question as to which structural domain of Leishmania GPIs is responsible for dramatic down-regulation of PKC-dependent transient c-fos expression, the chemically synthesized defined alkylacylglycerolipids domain of corresponding GPIs, and LPG and GIPLs isolated from Leishmania donovani, were evaluated for inhibition of PKC and c-fos expression in macrophages. The results presented here demonstrate that the unusual lipid domain of Leishmania GPIs is primarily responsible for inhibition of PKC-dependent transient c-fos expression. PMID- 12562867 TI - pp90RSK- and protein kinase C-dependent pathway regulates p42/44MAPK-induced LDL receptor transcription in HepG2 cells. AB - We have previously shown that different extracellular stimuli require signaling through the Raf/MEK/p42/44MAPK cascade to induce LDL receptor expression. The present studies were designed to delineate the molecular mechanisms underlying p42/44MAPK-induced LDL receptor transcription in HepG2-Delta Raf-1:ER cells, a modified HepG2 cell line in which the Raf-1/MEK/p42/44MAPK cascade can be specifically activated by anti-estradiol ICI182,780 in an agonist-specific manner. Using these cells, we show that: a) LDL receptor induction was reduced in reporter constructs containing mutation in either Sp1 or sterol-regulatory element-1 (SRE-1) sites, whereas inactivation of both sites abolished the induction; b) E1A, which inhibits CREB binding protein (CBP), a common activator of SRE-1 binding protein and Sp1, strongly repressed the induction; c) intracellular inhibition of the 90 kDa ribosomal S6 kinase (pp90RSK) cascade reduced LDL receptor induction; d) highly selective protein kinase C (PKC) inhibitors effectively abrogated the induction without affecting activation of pp90RSK; and e) overexpression of PKC beta significantly induced LDL receptor promoter activity. Taken together, these results demonstrate that pp90RSK and PKC beta are downstream effectors and Sp1, SRE-1 binding protein, and CBP are part of the transcriptional complex resulting in induction of LDL receptor expression in response to activation of the Raf/MEK/p42/44MAPK cascade. These findings identify for the first time a role for PKC beta in determining the specificity of p42/44MAPK signaling by participating with pp90RSK in regulating gene expression. PMID- 12562868 TI - Effects of dietary fat type and energy restriction on adipose tissue fatty acid composition and leptin production in rats. AB - To investigate whether dietary fatty acid (FA) composition and energy restriction (ER) interactively influence obese (ob) gene expression, rats consumed diets containing beef tallow, safflower, or fish oil ad libitum (AL) or at 60% AL intake. Circulating leptin concentrations were higher (P < 0.0001) after AL feeding, but were not influenced by dietary fat. ER decreased (P < 0.0001) weight gain and visceral adipose weight, which were positively correlated (r = 0.40 P < 0.001, r = 0.58 P < 0.0001) with circulating leptin levels. Visceral adipose ob mRNA levels were greater in animals fed unsaturated fats, particularly safflower oil, which had the highest ob mRNA levels. Circulating leptin levels did not parallel ob mRNA levels, except for the greater abundance detected in AL adipose in comparison to ER animals. In addition, visceral FA profiles reflected dietary fat source and were influenced by an interaction of dietary fat and energy. These data demonstrate that dietary fat, particularly from a plant or marine source, and ER interactively influence ob mRNA levels; however, alterations in ob mRNA do not confer changes in circulating leptin, with the exception of ER, which is a key determinant. Thus, dietary intake is an important regulator of leptin production; however, the significance of these modest changes in diet-induced obese animals requires further study. PMID- 12562869 TI - Circulating oxidized LDL forms complexes with beta2-glycoprotein I: implication as an atherogenic autoantigen. AB - Beta2-glycoprotein I (beta2-GPI) is a major antigen for antiphospholipid antibodies (Abs, aPL) present in patients with antiphospholipid syndrome (APS). We recently reported (J. Lipid Res., 42: 697, 2001; J. Lipid Res., 43: 1486, 2002) that beta2-GPI specifically binds to Cu2+-oxidized LDL (oxLDL) and that the beta2-GPI ligands are omega-carboxylated 7-ketocholesteryl esters. In the present study, we demonstrate that oxLDL forms stable and nondissociable complexes with beta2-GPI in serum, and that high serum levels of the complexes are associated with arterial thrombosis in APS. A conjugated ketone function at the 7-position of cholesterol as well as the omega-carboxyl function of the beta2-GPI ligands was necessary for beta2-GPI binding. The ligand-mediated noncovalent interaction of beta2-GPI and oxLDL undergoes a temperature- and time-dependent conversion to much more stable but readily dissociable complexes in vitro at neutral pH. In contrast, stable and nondissociable beta2-GPI-oxLDL complexes were frequently detected in sera from patients with APS and/or systemic lupus erythematodes. Both the presence of beta2-GPI-oxLDL complexes and IgG Abs recognizing these complexes were strongly associated with arterial thrombosis. Further, these same Abs correlated with IgG immune complexes containing beta2-GPI or LDL. Thus, the beta2 GPI-oxLDL complexes acting as an autoantigen are closely associated with autoimmune-mediated atherogenesis. PMID- 12562870 TI - Dietary sodium chloride restriction enhances aortic wall lipid storage and raises plasma lipid concentration in LDL receptor knockout mice. AB - This study aimed at measuring the influence of a low salt diet on the development of experimental atherosclerosis in moderately hyperlipidemic mice. Experiments were carried out on LDL receptor (LDLR) knockout (KO) mice, or apolipoprotein E (apoE) KO mice on a low sodium chloride diet (LSD) as compared with a normal salt diet (NSD). On LSD, the rise of the plasma concentrations of TG and nonesterified fatty acid (NEFA) was, respectively, 19% and 34% in LDLR KO mice, and 21% and 35% in apoE KO mice, and that of plasma cholesterol was limited to the LDLR KO group alone (15%). Probably due to the apoE KO severe hypercholesterolemia, the arterial inner-wall fat storage was not influenced by the diet salt content and was far more abundant in the apoE KO than in the LDLR KO mice. However, in the less severe hypercholesterolemia of the LDLR KO mice, lipid deposits on the LSD were greater than on the NSD. Arterial fat storage correlated with NEFA concentrations in the LDLR KO mice alone (n = 14, P = 0.0065). Thus, dietary sodium chloride restriction enhances aortic wall lipid storage in moderately hyperlipidemic mice. PMID- 12562871 TI - Plasma kinetics of a cholesterol-rich emulsion in subjects with or without coronary artery disease. AB - A cholesterol-rich emulsion (LDE) that resembles the LDL lipidic structure is taken-up by LDL receptors after intravenous injection by means of apolipoprotein E it acquires in the circulation and can be used to probe LDL metabolism. In this study, LDE was labeled with [14C]cholesteryl oleate and [3H]cholesterol and injected into 19 patients with coronary artery disease (CAD) and into 14 subjects without CAD to verify whether the kinetic behavior of the radioactive lipids is different in CAD. Blood was sampled over 24 h for radioactivity measurement after lipid extraction and separation by thin-layer chromatography. Fractional clearance rate (FCR, in h-1) of [14C]cholesteryl ester was not different in CAD and nonCAD expressed as median (25%; 75%): 0.08 (0.062; 0.134) h-1 versus 0.06 (0.04; 0.083) h-1, P = 0.167. However, [3H]cholesterol FCR was greater in CAD than in nonCAD (mean +/- SEM): 0.163 +/- 0.016 h-1 versus 0.077 +/- 0.014 h-1, P < 0.001. Esterification of the LDE [3H]cholesterol was also greater in CAD subjects than nonCAD at 10 h and 24 h after emulsion injection (P = 0.029 and 0.024 respectively). In conclusion, both removal from the plasma and esterification of the LDE-cholesterol were increased in CAD. These findings may contribute for unraveling pro-atherogenic mechanisms and the establishment of novel CAD markers. PMID- 12562872 TI - HDL regulates the displacement of hepatic lipase from cell surface proteoglycans and the hydrolysis of VLDL triacylglycerol. AB - We have previously shown that hepatic lipase (HL) is inactive when bound to purified heparan sulfate proteoglycans and can be liberated by HDL and apolipoprotein A-I (apoA-I), but not by LDL or VLDL. In this study, we show that HDL is also able to displace HL directly from the surface of the hepatoma cell line, HepG2, and Chinese hamster ovary cells stably overexpressing human HL. ApoA I is more efficient at displacing cell surface HL than is HDL, and different HDL classes vary in their ability to displace HL from the cell surface. HDL2s have a greater capacity to remove HL from the cell surface and intracellular compartments, as compared with the smaller HDL particles. The different HDL subclasses also uniquely affect the activity of the enzyme. HDL2 stimulates HL mediated hydrolysis of VLDL-triacylglycerol, while HDL3 is inhibitory. Inhibition of VLDL hydrolysis appears to result from a decreased interlipoprotein shuttling of HL between VLDL and the smaller, more dense HDL particles. This study suggests that high HDL2 levels are positively related to efficient triacylglycerol hydrolysis by their ability to enhance the liberation of HL into the plasma compartment and by a direct stimulation of VLDL-triacylglycerol hydrolysis. PMID- 12562873 TI - Fatty liver in familial hypobetalipoproteinemia: triglyceride assembly into VLDL particles is affected by the extent of hepatic steatosis. AB - Familial hypobetalipoproteinemia (FHBL) subjects may develop fatty liver. Liver fat was assessed in 21 FHBL with six different apolipoprotein B (apoB) truncations (apoB-4 to apoB-89) and 14 controls by magnetic resonance spectroscopy (MRS). Liver fat percentages were 16.7 +/- 11.5 and 3.3 +/- 2.9 (mean +/- SD) (P = 0.001). Liver fat percentage was positively correlated with body mass index, waist circumference, and areas under the insulin curves of 2 h glucose tolerance tests, suggesting that obesity may affect the severity of liver fat accumulation in both groups. Despite 5-fold differences in liver fat percentage, mean values for obesity and insulin indexes were similar. Thus, for similar degrees of obesity, FHBL subjects have more hepatic fat. VLDL triglyceride (TG)-fatty acids arise from plasma and nonplasma sources (liver and splanchnic tissues). To assess the relative contributions of each, [2H2]palmitate was infused over 12 h in 13 FHBL subjects and 11 controls. Isotopic enrichment of plasma free palmitate and VLDL-TG-palmitate was determined by mass spectrometry. Non-plasma sources contributed 51 +/- 15% in FHBL and 37 +/- 13% in controls (P = 0.02). Correlations of liver fat percentage and percent VLDL-TG-palmitate from liver were r = 0.89 (P = 0.0001) for FHBL subjects and r = 0.69 (P = 0.01) for controls. Thus, apoB truncation-producing mutations result in fatty liver and in altered assembly of VLDL-TG. PMID- 12562874 TI - Effects of oleic acid and its congeners, elaidic and stearic acids, on the structural properties of phosphatidylethanolamine membranes. AB - Fatty acid derivatives are abundant in biological membranes, mainly as components of phospholipids and cholesterol esters. Their presence, free or bound to phospholipids, modulates the lipid membrane behavior. The present study shows the differential influence of the C-18 fatty acids (FAs), oleic, elaidic, and stearic acids on the structural properties of phosphatidylethanolamine (PE). X-ray diffraction of PE-FA systems demonstrated that oleic acid (OA) produced important concentration-dependent alterations of the lipid membrane structure: it induced reductions of up to 20-23 degrees C in the lamellar-to-hexagonal transition temperature of 1-palmitoyl-2-oleoyl PE and dielaidoyl PE and regulated the dimensions of the hexagonal lattice. In contrast, elaidic and stearic acids did not markedly alter the phospholipid mesomorphism. The above effects were attributed to the different "molecular shape" of OA (with a kink at the middle of the molecule) with respect to their congeners, elaidic and stearic acids. The effects of free fatty acids (FFAs) on membrane structure are relevant for several reasons: i) some biological membranes contain very high levels of FFAs. ii) Mediterranean diets with high OA intake have been shown to exert protective effects against tumoral and hypertensive pathologies. iii) FFA derivatives have been developed as antitumoral and antihypertensive drugs. PMID- 12562875 TI - Differential modulation of Toll-like receptors by fatty acids: preferential inhibition by n-3 polyunsaturated fatty acids. AB - Human subjects consuming fish oil showed a significant suppression of cyclooxygenase-2 (COX-2) expression in blood monocytes when stimulated in vitro with lipopolysaccharide (LPS), an agonist for Toll-like receptor 4 (TLR4). Results with a murine monocytic cell line (RAW 264.7) stably transfected with COX 2 promoter reporter gene also demonstrated that LPS-induced COX-2 expression was preferentially inhibited by docosahexaenoic acid (DHA, C22:6n-3) and eicosapentaenoic acid (EPA, C20:5n-3), the major n-3 polyunsaturated fatty acids (PUFAs) present in fish oil. Additionally, DHA and EPA significantly suppressed COX-2 expression induced by a synthetic lipopeptide, a TLR2 agonist. These results correlated with the preferential suppression of LPS- or lipopeptide induced NF kappa B activation by DHA and EPA. The target of inhibition by DHA is TLR itself or its associated molecules, but not downstream signaling components. In contrast, COX-2 expression by TLR2 or TRL4 agonist was potentiated by lauric acid, a saturated fatty acid. These results demonstrate that inhibition of COX-2 expression by n-3 PUFAs is mediated through the modulation of TLR-mediated signaling pathways. Thus, the beneficial or detrimental effects of different types of dietary fatty acids on the risk of the development of many chronic inflammatory diseases may be in part mediated through the modulation of TLRs. PMID- 12562876 TI - Autoimmune response to advanced glycosylation end-products of human LDL. AB - Advanced glycosylation end-products (AGEs) are believed to play a significant role in the development of vascular complications in diabetic patients. One such product, AGE-LDL, has been shown to be immunogenic. In this report, we describe the isolation and characterization of human AGE-LDL antibodies from the sera of seven patients with Type 1 diabetes by affinity chromatography using an immobilized AGE-LDL preparation that contained primarily the AGE N epsilon (carboxymethyl)lysine (CML, 14.6 mmol/mol lysine), and smaller amounts of N epsilon (carboxyethyl)lysine (CEL, 2.7 mmol/mol lysine). The isolated antibodies were predominantly IgG of subclasses 1 and 3, and considered proinflammatory because of their ability to promote Fc gamma R-mediated phagocytosis and to activate complement. We determined dissociation constants (Kd) for the purified antibodies. The average Kd values (4.76 +/- 2.52 x 10(-9) mol/l) indicated that AGE-LDL antibodies are of higher avidity than oxidized LDL antibodies measured previously (Kd = 1.53 +/- 07 x 10(-8) ml/l), but of lower avidity than rabbit polyclonal LDL antibodies (Kd = 9.34 x 10(-11)). Analysis of the apolipoprotein B rich lipoproteins isolated with polyethylene glycol-precipitated antigen-antibody complexes from the same patients showed the presence of both CML and CEL, thus confirming that these two modifications are recognized by human autoantibodies. A comparative study of the reactivity of purified AGE-LDL antibodies with CML-LDL and CML-serum albumin showed no cross-reactivity. PMID- 12562877 TI - Salivary biochemistry in Buffalo: the legacy of Michael J. Levine. PMID- 12562878 TI - Resolution of inflammation: a new paradigm for the pathogenesis of periodontal diseases. AB - The periodontal diseases are infectious diseases caused by predominantly Gram negative bacteria. However, as our understanding of the pathogenesis of the periodontal diseases grows, it is becoming clear that most of the tissue damage that characterizes periodontal disease is caused by the host response to infection, not by the infectious agent directly. Investigation into the mechanism of action of host-mediated tissue injury has revealed that the neutrophil plays an important role in destruction of host tissues. In this paper, we review the biochemical pathways and molecular mediators that are responsible for regulation of the inflammatory response in diseases such as periodontitis, with a focus on lipid mediators of inflammation. Pro-inflammatory mediators, such as prostaglandins and leukotrienes, are balanced by counter-regulatory signals provided by a class of molecules called lipoxins. The role of lipoxins in the control and resolution of inflammation is discussed, as is the possibility of the development of new therapeutic strategies for the control and prevention of neutrophil-mediated tissue injury in inflammatory diseases like periodontitis. PMID- 12562879 TI - Transient co-localization of c-Jun N-terminal kinase and c-Jun with heat shock protein 70 in pulp cells during apoptosis. AB - The c-Jun N-terminal kinase (JNK) pathway and heat-shock proteins (HSPs) are involved in stress-induced apoptosis. Here we examined the association of JNK, c Jun, and anti-apoptotic HSPs with pulp apoptosis during wound healing. In normal pulp, c-Jun was activated only in a few pulp cells, but JNK was not. HSP70 was expressed in the cytoplasm of pulp cells. One day after injury, active JNK and c Jun were detected in apoptotic pulp cells, whereas HSP70 was detected in non apoptotic cells. We also found the translocation of HSP70 into nuclei of pulp cells, and co-localization with active JNK and c-Jun. Four days after injury, active JNK and c-Jun disappeared in pulp cells, and HSP70 was relocalized from nuclei to the cytoplasm. These results suggest that the JNK pathway may be one of the compartments inducing apoptosis in pulp cells, and that HSP70 may have an inhibitory role in the apoptosis of pulp cells during wound healing. PMID- 12562880 TI - Production of colony-stimulating factor in human dental pulp fibroblasts. AB - Class II major histocompatilibity complex (MHC)-expressing cells are usually distributed in dental pulp, and it was postulated that the colony-stimulating factor (CSF) derived from dental pulp fibroblasts contributes to the migration of class II MHC-expressing cells into pulp tissue. This study aimed to investigate the CSF production of human dental pulp fibroblasts. In pulp tissue sections, granulocyte (G)-CSF was detected from normal teeth, while G-CSF, macrophage (M) CSF, and granulocyte-macrophage (GM)-CSF were detected from teeth with dentinal caries. In cultured dental pulp fibroblasts, G-CSF was detected by immunostaining, immunoprecipitation, and ELISA, and mRNAs of G-CSF, M-CSF, and GM CSF were detected by RT-PCR. The dental pulp fibroblasts cultured with TNF-alpha were found to increase the G-CSF expression and to produce M-CSF and GM-CSF. These findings suggest that dental pulp fibroblasts usually produce G-CSF. In the presence of TNF-alpha, dental pulp fibroblast express M-CSF and GM-CSF. PMID- 12562881 TI - Destruction of parotid glands affects nitrate and nitrite metabolism. AB - The role of salivary glands in nitrate and nitrite metabolism is poorly understood. The aim of the present study was to investigate the effect of parotid gland ablation on dynamic metabolism of nitrate and nitrite in miniature pigs. The parotid glands of 5 healthy miniature pigs were bilaterally ablated by methyl violet. Concentrations of nitrate and nitrite of whole saliva, serum, and urine samples were analyzed by high-performance liquid chromatography. Results showed that bilateral ablation of the parotid glands led to a significant decrease of nitrate secretion from blood to saliva (P < 0.05) and thus low nitrite levels. Dysfunction of the parotid glands temporarily increased the systemic level of nitrate in miniature pigs after nitrate loading. This study suggests that the parotid glands play an important role in the balance of nitrate and nitrite levels in both whole saliva and the body. PMID- 12562882 TI - Functional heterogeneity in the superior head of the human lateral pterygoid. AB - The activity of the superior head of the human lateral pterygoid muscle (SHLP) is controversial. Given the non-parallel alignment of some SHLP fibers, the SHLP may be capable of differential activation. The aims were to clarify SHLP activity patterns in relation to location within SHLP. In 18 subjects, SHLP single motor units were intramuscularly recorded at computer-tomography-verified sites during horizontal (e.g., protrusion) and vertical (e.g., opening) jaw tasks (recorded by a jaw-tracking device) and at resting postural jaw position. None of 92 units was active at the resting postural position. Medially located units (21) showed activity during contralateral movement, protrusion, and opening; 5 were also active on jaw closing. There was a significant association between unit location and the number of units active during vertical tasks (i.e., jaw closing and clenching). Analysis of the data suggests differential activation within SHLP and raises the possibility of functional heterogeneity within SHLP. PMID- 12562883 TI - Sex differences in masticatory muscle pain after chewing. AB - Neither the etiology of muscle-related temporomandibular disorders (TMD) nor the reason for the disproportionate number of women suffering from these disorders is well-established. We tested the hypothesis that physiologically relevant exercise (i.e., chewing bubble gum for 6 min) increases masticatory muscle pain in patients, but not in asymptomatic control subjects, and that female patients experience a significantly greater increase than males. Chewing increased pain in both female and male patients and, unexpectedly, also in female control subjects. One hour after chewing, the pain remained above pre-test levels for female patients but not for the other groups. Thus, sex differences in chewing-induced pain were found in control subjects but not as hypothesized in patients. Because chewing-induced masticatory muscle pain was significantly greater in female control subjects than in males, and persisted longer in female patients than in males, these results suggest greater susceptibility in women. PMID- 12562884 TI - Construct and predictive validity of clinical caries diagnostic criteria assessing lesion activity. AB - Even though there is no "gold standard" for determining caries lesion activity, it is nonetheless possible to evaluate the validity of such diagnostic measures. The aim of this study was to estimate the construct and predictive validity of caries lesion activity assessments by means of their ability to reflect known effects of fluoride on caries. A three-year trial of the effect of daily supervised brushing with fluoride toothpaste was carried out among 273 12-year old children. All children were examined clinically according to diagnostic criteria for activity assessment. The relative risk (fluoride vs. control) for caries lesion transitions among diagnostic categories was calculated. Fluoride inhibited progression of caries at all stages of lesion formation while at the same time enhancing lesion regression. The effects were most pronounced for active non-cavitated lesions. It is concluded that the clinical diagnostic criteria have construct and predictive validity for the assessment of caries lesion activity. PMID- 12562885 TI - Association of edentulism and diet and nutrition in US adults. AB - Dental status and its relationship to diet and nutritional status have been little explored. In this study of a representative sample of the US civilian, non institutionalized population (NHANES III), we predicted that the intake of nutritious foods, dietary fiber, and levels of biochemical analytes would be lower, even after adjusting for potential social and behavioral factors, among those who were edentulous and wore complete dentures than for those who had all their natural teeth. Multivariate analyses indicated that intake of carrots and tossed salads among denture-wearers was, respectively, 2.1 and 1.5 times less than for the fully dentate (p < 0.0001), and dietary fiber intake was 1.2 times less (p < 0.05). Serum levels of beta carotene (9.8 microg/dL), folate (4.7 ng/dL), and vitamin C (0.87 mg/dL) were also lower among denture-wearers (p < 0.05). Intakes of some nutrient-rich foods and beta carotene, folate, and vitamin C serum levels were significantly lower in denture-wearers. PMID- 12562886 TI - Adjustment of the occlusal vertical dimension in the bite-raised guinea pig. AB - In humans, the inappropriate occlusal vertical dimension (OVD) causes several orofacial disorders, such as bruxism and pain in the masticatory muscles and temporomandibular joint. However, it is still unclear how strictly the OVD is adjusted. To answer this question, we studied the temporal change of the OVD in bite-raised young guinea pigs. The OVD was raised by fixation of a bite-raising appliance to the lower incisors, and increased by either 3 or 1.5 mm at the first molars. After the space produced between the upper and lower molars was filled within 10 days due to eruption of the molars, the appliance was removed. In the bite-raised animals, the raised OVD was reduced until it attained that observed in the control animals, after which the OVD increased according to cranial growth. These results show that the OVD is developmentally changed and strictly controlled. PMID- 12562887 TI - Rabbit calvarial wound healing by means of seeded Caprotite scaffolds. AB - Autologous bone is the most successful bone-grafting material; however, limited supply and donor site morbidity are problematic. Synthetic bone substitutes are effective, but healing is slow and unpredictable. Osseous wound healing may be enhanced if bone substitutes are combined with autologous bone marrow cells. To test this hypothesis, we created 40 calvarial defects in 20 12-week-old New Zealand White rabbits, divided into four groups: (1) unrepaired controls, (2) autologous bone grafts, (3) unseeded Caprotite (a polymer-ceramic composite) grafts, and (4) Caprotite grafts seeded with autologous bone marrow stromal cells. CT scans were obtained at 0, 6, and 12 weeks post-operatively, and defects were harvested for histology. Defects repaired with autologous bone had significantly (p < 0.05) more bone than the other three groups, although seeded Caprotite defects showed different wound-healing sequelae. Results suggest that seeded Caprotite scaffolds did not significantly enhance osseous defect healing compared with controls. PMID- 12562888 TI - Four-year water degradation of total-etch adhesives bonded to dentin. AB - Resin-dentin bonds degrade over time. The objective of this study was to evaluate the influence of variables like hybridization effectiveness and diffusion/elution of interface components on degradation. Hypotheses tested were: (1) There is no difference in degradation over time between two- and three-step total-etch adhesives; and (2) a composite-enamel bond protects the adjacent composite-dentin bond against degradation. The micro-tensile bond strength (microTBS) to dentin of 2 three-step total-etch adhesives was compared with that of 2 two-step total-etch adhesives after 4 years of storage in water. Quantitative and qualitative failure analyses were conducted correlating Fe-SEM and TEM. Indirect exposure to water did not significantly reduce the microTBS of any adhesive, while direct exposure resulted in a significantly reduced microTBS of both two-step adhesives. It is concluded that resin bonded to enamel protected the resin-dentin bond against degradation, while direct exposure to water for 4 years affected bonds produced by two-step total-etch adhesives. PMID- 12562889 TI - Hybridization efficiency of the adhesive/dentin interface with wet bonding. AB - Although it is generally proposed that dentin bonding results from adhesive infiltration of superficially demineralized dentin, it is not clear how well the resin monomers seal the dentin collagen fibrils under wet bonding conditions. The aim of this study was to determine the quality and molecular structure of adhesive/dentin (a/d) interfaces formed with wet bonding as compared with adhesive-infiltrated demineralized dentin (AIDD) produced under controlled conditions (optimum hybrid). From each extracted, unerupted human 3rd molar, one fraction was demineralized, dehydrated, and infiltrated with Single Bond (SB) adhesive under optimum conditions; the remaining, adjacent fraction was treated with SB by wet bonding. AIDD and a/d interface sections were stained with Goldner's trichrome; corresponding sections were analyzed with micro-Raman spectroscopy. The histomorphologic and spectroscopic results suggest that, under wet bonding, the a/d interface is a porous collagen web infiltrated primarily by the hydrolytically unstable HEMA. PMID- 12562890 TI - Effect of smear layer on root demineralization adjacent to resin-modified glass ionomer. AB - The cariostatic effect of resin-modified glass ionomer (RMGI) on secondary root caries is well-documented. However, this beneficial effect may be dependent upon the mode of cavity surface treatment. To investigate this relationship, we studied 4 cavity surface treatments prior to the placement of RMGI: no treatment (None), polyacrylic acid (PAA), phosphoric acid (H(3)PO(4)), and Scotchbond Multi Purpose adhesive (SMP) as a control. Specimens were aged for two weeks in synthetic saliva, thermocycled, and subjected to an artificial caries challenge (pH 4.4). Polarized light microscopy (PLM) and microradiography (MRG) showed significantly less demineralization with the H(3)PO(4) cavity surface treatment as revealed by ANOVA and Tukey's multiple comparisons (p < or = 0.05). Dentin fluoride profiles determined by electron probe microanalysis (EPMA) supported PLM and MRG findings. It may be concluded that removal of the smear layer with phosphoric acid provides significantly enhanced resistance to secondary root caries formation adjacent to RMGI restorations. PMID- 12562891 TI - The actions of anandamide on rat superficial medullary dorsal horn neurons in vitro. AB - Whole-cell patch-clamp recordings were made from neurons in the trigeminal nucleus caudalis and trigeminal ganglion, in vitro, to investigate the cellular actions of the endogenous cannabinoid, anandamide. Anandamide has been shown to act through both the cannabinoid receptor 1 (CB1) and the vanilloid receptor 1 (VR1). Anandamide (30 microM) caused a 54 % increase in the rate of miniature excitatory post-synaptic currents (mEPSCs), without affecting their amplitude. The effect of anandamide was blocked by the VR1 antagonist capsazepine (20 microM), but not by the CB1-specific antagonist AM251 (3 microM). Application of the VR1 receptor agonist capsaicin (300 nM) caused a 4200 % increase in the mEPSC rate. In dissociated trigeminal ganglion neurons, both anandamide and capsaicin caused an outward current in neurons that were voltage clamped at +40 mV. The maximal outward current produced by anandamide (EC50, 10 microM) was 45 % of that produced by capsaicin (10 microM). Co-application of the VR1 antagonist capsazepine (30 microM) completely reversed the effects of both capsaicin and anandamide. The anandamide transport inhibitor, AM404 (30 microM) caused a 40 % increase in mEPSC rate in the slice preparation and an outward current in dissociated neurons. The latter current was reversed by the VR1 antagonist iodoresiniferatoxin (1 microM). The fatty acid amide hydrolase (FAAH) inhibitors phenylmethylsulfonyl fluoride (PMSF) (20 microM) and OL53 (1 microM) did not enhance the effect of anandamide in either the slice or dissociated neuron preparations. These results suggest that within the superficial medullary dorsal horn, anandamide (30 microM) acts presynaptically to enhance the release of glutamate via activation of the VR1 receptor. PMID- 12562892 TI - Matrix volume measurements challenge the existence of diazoxide/glibencamide sensitive KATP channels in rat mitochondria. AB - A mitochondrial sulphonylurea-sensitive, ATP-sensitive K+ channel (mitoKATP) that is selectively inhibited by 5-hydroxydecanoate (5-HD) and activated by diazoxide has been implicated in ischaemic preconditioning. Here we re-evaluate the evidence for the existence of this mitoKATP by measuring changes in light scattering (A520) in parallel with direct determination of mitochondrial matrix volumes using 3H2O and [14C]sucrose. Incubation of rat liver and heart mitochondria in KCl medium containing Mg2+ and inorganic phosphate caused a decrease in light scattering over 5 min, which was accompanied by a small (15-30 %) increase in matrix volume. The presence of ATP or ADP in the buffer from the start greatly inhibited the decline in A520, whilst addition after a period of incubation (1-5 min) induced a rapid increase in A520, especially in heart mitochondria. Neither response was accompanied by a change in matrix volume, as measured isotopically. However, the effects of ATP and ADP on A520 were abolished by carboxyatractyloside and bongkrekic acid, inhibitors of the adenine nucleotide translocase (ANT) that lock the transporter in two discrete conformations and cause distinct changes in A520 in their own right. These data suggest that rather than matrix volume changes, the effects of ATP and ADP on A520 reflect changes in mitochondrial shape induced by conformational changes in the ANT. Furthermore, we were unable to demonstrate either a decrease in A520 or increase in matrix volume with a range of ATP-sensitive K+ channel openers such as diazoxide. Nor did glibencamide or 5-HD cause any reduction of matrix volume, whereas the K+ ionophore valinomycin (0.2 nM), produced a 10-20 % increase in matrix volume that was readily detectable by both techniques. Our data argue against the existence of a sulphonylurea-inhibitable mitoKATP channel. PMID- 12562893 TI - Creatine kinase injection restores contractile function in creatine-kinase deficient mouse skeletal muscle fibres. AB - Viable genetically engineered animals generally exhibit adaptations to the altered genotype, which may mask the role of the protein of interest. We now describe a novel method by which the direct effects of the altered genotype can be distinguished from secondary adaptive changes in isolated adult skeletal muscle cells. We studied contractile function and intracellular Ca2+ handling in single skeletal muscle fibres that are completely deficient of creatine kinase (CK; CK-/-) before and after microinjection of purified CK (injected together with the fluorescent Ca2+ indicator indo-1). The mean total CK activity after CK injection was estimated to be approximately 4 mM s-1, which is approximately 5 % of the activity in wild-type muscle fibres. After CK injection, CK-/- fibres approached the wild-type phenotype in several aspects: (a) the free myoplasmic [Ca2+] ([Ca2+]i) increased and force showed little change during a period of high intensity stimulation (duty cycle, i.e. tetanic duration divided by tetanic interval = 0.67); (b) [Ca2+]i did not decline during a brief (350 ms) tetanus; (c) during low-intensity fatiguing stimulation (duty cycle = 0.14), tetanic [Ca2+]i increased over the first 10 tetani, and thereafter it decreased; (d) tetanic [Ca2+]i and force did not display a transient reduction in the second tetanus of low-intensity fatiguing stimulation. Conversely, tetanic force in the unfatigued state was lower in CK-/- than in wild-type fibres, and this difference persisted after CK injection. Injection of inactivated CK had no obvious effect on any of the measured parameters. In conclusion, microinjection of CK into CK-/- fibres markedly restores many, but not all, aspects of the wild-type phenotype. PMID- 12562895 TI - Effects of dynamic exercise intensity on the activation of hormone-sensitive lipase in human skeletal muscle. AB - It has been proposed that hormone-sensitive lipase (HSL) regulates intramuscular triacylglycerol hydrolysis in skeletal muscle. The primary purpose of this study was to examine the early activation of HSL and the changes in the putative intramuscular and hormonal regulators of HSL activity at various aerobic exercise intensities. Eight male subjects cycled for 10 min at power outputs corresponding to 30, 60 and 90 % peak oxygen uptake (VO(2,peak)). Muscle samples were obtained at rest and following 1 and 10 min of exercise. Intramuscular triacylglycerol (mean +/- S.E.M.: 24.3 +/- 2.3 mmol (kg dry mass (DM))(-1)), long-chain fatty acyl CoA (13.9 +/- 1.4 micromol (kg DM)(-1)) and HSL activity (1.87 +/- 0.07 mmol min(-1) (kg DM)(-1))) were not different between trials at rest. HSL activity increased at 1 min of exercise at 30 and 60 % VO(2,peak), and to a greater extent at 90 % VO(2,peak). HSL activity remained elevated after 10 min of exercise at 30 and 60 % VO(2,peak), and decreased at 90 % VO(2,peak) from the rates observed at 1 min (1 min: 3.41 +/- 0.3 mmol min(-1) (kg DM)-1; 10 min: 2.92 +/- 0.26 mmol min(-1) (kg DM)(-1)), P < 0.05). There were no effects of exercise power output or time on long-chain fatty acyl CoA content. At 90 % VO(2,peak), skeletal muscle contents of ATP and phosphocreatine were decreased (P < 0.05), and free ADP and free AMP were increased (P < 0.05) during exercise. No changes in these metabolites occurred at 30 % VO(2,peak) and only modest changes were observed at 60 % VO(2,peak). Plasma adrenaline increased (P < 0.05) during exercise at 90 % VO(2,peak) only. These data suggest that a factor related to the onset of exercise (e.g. Ca2+) activates HSL early in exercise. Given the activation of HSL early in exercise, at a time when intramuscular triacylglycerol hydrolysis and fat oxidation are considered to be negligible, we propose that the control of intramuscular triacylglycerol hydrolysis is not solely related to the level of HSL activation, but must also be regulated by postactivational factors. PMID- 12562894 TI - Protein kinase C is necessary for recovery from the thyrotropin-releasing hormone induced r-ERG current reduction in GH3 rat anterior pituitary cells. AB - The biochemical cascade linking activation of phospholipase C-coupled thyrotropin releasing hormone (TRH) receptors to rat ERG (r-ERG) channel modulation was studied in situ using perforated-patch clamped adenohypophysial GH3 cells and pharmacological inhibitors. To check the recent suggestion that Rho kinase is involved in the TRH-induced r-ERG current suppression, the hormonal effects were studied in cells pretreated with the Rho kinase inhibitors Y-27632 and HA-1077. The TRH-induced r-ERG inhibition was not significantly modified in the presence of the inhibitors. Surprisingly, the hormonal effects became irreversible in the presence of HA-1077 but not in the presence of the more potent Rho kinase inhibitor Y-27632. Further experiments indicated that the effect of HA-1077 correlated with its ability to inhibit protein kinase C (PKC). The hormonal effects also became irreversible in cells in which PKC activity was selectively impaired with GF109203X, Go6976 or long-term incubation with phorbol esters. Furthermore, the reversal of the effects of TRH, but not its ability to suppress r-ERG currents, was blocked if diacylglycerol generation was prevented by blocking phospholipase C activity with U-73122. Our results suggest that a pathway involving an as yet unidentified protein kinase is the main cause of r ERG inhibition in perforated-patch clamped GH3 cells. Furthermore, they demonstrate that although not necessary to trigger the ERG current reductions induced by TRH, an intracellular signal cascade involving phosphatidylinositol 4,5-bisphosphate hydrolysis by phospholipase C, activation of an alpha/betaII conventional PKC and one or more dephosphorylation steps catalysed by protein phosphatase 2A, mediates recovery of ERG currents following TRH withdrawal. PMID- 12562896 TI - Hydrogen-peroxide-induced toxicity of rat striatal neurones involves activation of a non-selective cation channel. AB - Striatal neurones are particularly vulnerable to hypoxia/ischaemia-induced damage, and free radicals are thought to be prime mediators of this neuronal destruction. It has been shown that hydrogen peroxide (H2O2), through the production of free radicals, induces rat insulinoma cell death by activation of a non-selective cation channel, which leads to irreversible cell depolarization and unregulated Ca2+ entry into the cell. In the study presented here, we demonstrate that a subpopulation of striatal neurones (medium spiny neurones) is depolarized by H2O2 through the production of free radicals. Cell-attached recordings from rat cultured striatal neurones demonstrate that exposure to H2O2 opens a large conductance channel that is characterized by extremely long open times (seconds). Inside-out recordings show that cytoplasmically applied beta-nicotinamide adenine dinucleotide activates a channel with little voltage dependence, a linear current voltage relationship and a single-channel conductance of between 70 and 90 pS. This channel is permeable to Na+, K+ and Ca2+ ions. Fura-2 imaging from cultured striatal neurones reveals that H2O2 exposure induces a biphasic intracellular Ca2+ increase in a subpopulation of neurones, the second, later phase resulting in Ca2+ overload. This later component of the Ca2+ response is dependent on the presence of extracellular Ca2+ and is independent of synaptic activity or voltage gated Ca2+ channel opening. Consequently, this channel may be an important contributor to free radical-induced selective striatal neurone destruction. These results are remarkably similar to those observed for insulinoma cells and suggest that this family of non-selective cation channels has a widespread distribution in mammalian tissues. PMID- 12562897 TI - Arm blood flow and oxygenation on the transition from arm to combined arm and leg exercise in humans. AB - The cardiovascular response to exercise with several groups of skeletal muscle implies that work with the legs may reduce arm blood flow. This study followed arm blood flow (Yarm) and oxygenation on the transition from arm cranking (A) to combined arm and leg exercise (A+L). Seven healthy male subjects performed A at approximately 80 % of maximum work rate (Wmax) and A at ~80 % Wmax combined with L at approximately 60 % Wmax. A transition trial to volitional exhaustion was performed where L was added after 2 min of A. The Yarm was determined by constant infusion thermodilution in the axillary vein and changes in biceps muscle oxygenation were measured with near-infrared spectroscopy. During A+L Yarm was lowered by 0.38 +/- 0.06 l min-1 (10.4 +/- 3.3 %, P < 0.05) from 2.96 +/- 1.54 l min-1 during A. Total (HbT) and oxygenated haemoglobin (HbO2) concentrations were also lower. During the transition from A to A+L Yarm decreased by 0.22 +/- 0.03 l min-1 (7.9 +/- 1.8 %, P < 0.05) within 9.6 +/- 0.2 s, while HbT and HbO2 decreased similarly within 30 +/- 2 s. At the same time mean arterial pressure and arm vascular conductance also decreased. The data demonstrate reduction in blood flow to active skeletal muscle during maximal whole body exercise to a degree that arm oxygen uptake and muscle tissue oxygenation are compromised. PMID- 12562899 TI - Role of the transverse-axial tubule system in generating calcium sparks and calcium transients in rat atrial myocytes. AB - Cardiac atrial cells lack a regular system of transverse tubules like that in cardiac ventricular cells. Nevertheless, many atrial cells do possess an irregular internal transverse-axial tubular system (TATS). To investigate the possible role of the TATS in excitation-contraction coupling in atrial myocytes, we visualized the TATS (labelled with the fluorescent indicator, Di-8-ANEPPS) simultaneously with Ca2+ transients and/or Ca2+ sparks (fluo-4). In confocal transverse linescan images of field-stimulated cells, whole-cell Ca2+ transients had two morphologies: 'U-shaped' transients and irregular or 'W-shaped' transients with a varying number of points of origin of the Ca2+ transient. About half (54 %, n =289 cells, 13 animals) of the cells had a TATS. Cells with TATS had a larger mean diameter (13.2 +/- 2.8 microm) than cells without TATS (11.7 +/ 2.0 microm) and were more common in the left atrium (n = 206 cells; left atrium: 76 with TATS, 30 without TATS; right atrium: 42 with TATS, 58 without TATS). Simultaneous measurement of Ca2+ sparks and sarcolemmal structures showed that cells without TATS had U-shaped transients that started at the cell periphery, and cells with TATS had W-shaped transients that began simultaneously at the cell periphery and the TATS. Most (82 out of 102 from 31 cells) 'spontaneous' (non depolarized) Ca2+ sparks occurred within 1 microm of a sarcolemmal structure (cell periphery or TATS), and 33 % occurred within 1 pixel (0.125 microm). We conclude that the presence of a sarcolemmal membrane either at the cell periphery or in the TATS in close apposition to the sarcoplasmic reticulum is required for the initiation of an evoked Ca2+ transient and for spontaneous Ca2+ sparks. PMID- 12562898 TI - Polarized distribution of HCO3- transport in human normal and cystic fibrosis nasal epithelia. AB - The polarized distribution of HCO3- transport was investigated in human nasal epithelial cells from normal and cystic fibrosis (CF) tissues. To test for HCO3- transport via conductive versus electroneutral Cl-/HCO3- exchange (anion exchange, AE) pathways, nasal cells were loaded with the pH probe 2',7' bis(carboxyethyl)-5(6)-carboxyfluorescein and mounted in a bilateral perfusion chamber. In normal, but not CF, epithelia, replacing mucosal Cl- with gluconate caused intracellular pH (pHi) to increase, and the initial rates (Delta pH min-1) of this increase were modestly augmented (approximately 26 %) when normal cells were pretreated with forskolin (10 microM). Recovery from this alkaline shift was dependent on mucosal Cl-, was insensitive to the AE inhibitor 4,4' diisothiocyanatodihydrostilbene-2,2'-disulfonic acid (H2DIDS; 1.5 mM), but was sensitive to the cystic fibrosis transmembrane conductance regulator (CFTR) channel inhibitor diphenylamine-2-carboxylate (DPC; 100 microM). In contrast, removal of serosal Cl- caused pHi to alkalinize in both normal and CF epithelia. Recovery from this alkaline challenge was dependent on serosal Cl- and blocked by H2DIDS. Additional studies showed that serosally applied Ba2+ (5.0 mM) in normal, but not CF, cells induced influx of HCO3- across the apical membrane that was reversibly blocked by mucosal DPC. In a final series of studies, normal and CF cells acutely alkaline loaded by replacing bilateral Krebs bicarbonate Ringer (KBR) with Hepes-buffered Ringer solution exhibited basolateral, but not apical, recovery from an alkaline challenge that was dependent on Cl-, independent of Na+ and blocked by H2DIDS. We conclude that: (1) normal, but not CF, nasal epithelia have a constitutively active DPC-sensitive HCO3- influx/efflux pathway across the apical membrane of cells, consistent with the movement of HCO3- via CFTR; and (2) both normal and CF nasal epithelia have Na+-independent, H2DIDS-sensitive AE at their basolateral domain. PMID- 12562901 TI - The quality of maximum likelihood estimates of ion channel rate constants. AB - Properties of maximum likelihood estimators of rate constants for channel mechanisms are investigated, to see what can and cannot be inferred from experimental results. The implementation of the HJCFIT method is described; it maximises the likelihood of an entire sequence of apparent open and shut times, with the rate constants in a specified reaction mechanism as free parameters. The exact method for missed brief events is used. Several methods for testing the quality of the fit are described. The distributions of rate constants, and correlations between them, are investigated by doing sets of 1000 fits to simulated experiments. In a standard nicotinic receptor mechanism, all nine free rate constants can be estimated even from one single channel recording, as long as the two binding sites are independent, even when the number of channels in the patch is not known. The estimates of rate constants that apply to diliganded channels are robust; good estimates can be obtained even with erroneous assumptions (e.g. about the value of a fixed rate constant or the independence of sites). Rate constants that require distinction between the two sites are less robust, and require that an EC50 be specified, or that records at two concentrations be fitted simultaneously. Despite the complexity of the problem, it appears that there exist two solutions with very similar likelihoods, as in the simplest case. The hazards that result from this, and from the strong positive correlation between estimates of opening and shutting rates, are discussed. PMID- 12562900 TI - Properties of the human muscle nicotinic receptor, and of the slow-channel myasthenic syndrome mutant epsilonL221F, inferred from maximum likelihood fits. AB - The mechanisms that underlie activation of nicotinic receptors are investigated using human recombinant receptors, both wild type and receptors that contain the slow channel myasthenic syndrome mutation, epsilonL221F. The method uses the program HJCFIT, which fits the rate constants in a specified mechanism directly to a sequence of observed open and shut times by maximising the likelihood of the sequence with exact correction for missed events. A mechanism with two different binding sites was used. The rate constants that apply to the diliganded receptor (opening, shutting and total dissociation rates) were estimated robustly, being insensitive to the exact assumptions made during fitting, as expected from simulation studies. They are sufficient to predict the main physiological properties of the receptors. The epsilonL221F mutation causes an approximately 4 fold reduction in dissociation rate from diliganded receptors, and a smaller increase in opening rate and mean open time. These are sufficient to explain the approximately 6-fold slowing of decay of miniature synaptic currents seen in patients. The distinction between the two binding sites was less robust, the estimates of rate constants being dependent to some extent on assumptions, e.g. whether an extra short-lived shut state was included or whether the EC50 was constrained. The results suggest that the two binding sites differ by roughly 10 fold in the affinity of the shut receptor for ACh in the wild type, and that in the epsilonL221F mutation the lower affinity is increased so the sites become more similar. PMID- 12562902 TI - Mechanisms of cold sensitivity of paramyotonia congenita mutation R1448H and overlap syndrome mutation M1360V. AB - Missense mutations of the human skeletal muscle voltage-gated Na+ channel (hSkM1) cause a variety of neuromuscular disorders. The mutation R1448H results in paramyotonia congenita and causes cold-induced myotonia with subsequent paralysis. The mutation M1360V causes an overlapping syndrome with both K+ induced muscle weakness and cold-induced myotonia. The molecular mechanisms of the temperature dependence of these disorders are not well understood. Therefore we investigated physiological parameters of these Na+ channel mutations at different temperatures. Channel proteins were recombinantly expressed in human embryonic kidney cells and studied electrophysiologically, using the whole-cell patch-clamp technique. We compared the wild-type (WT) channel with both mutants at different temperatures. Both mutations had slower inactivation and faster recovery from inactivation compared to WT channels. This effect was more pronounced at the R1448H mutation, leading to a larger depolarization of the cell membrane causing myotonia and paralysis. The voltage dependence of activation of R1448H was shifted to more negative membrane potentials at lower temperature but not at the M1360V mutation or in the WT. The window current by mutation R1448H was increased at lower temperatures. The results of this study may explain the stronger cold-induced clinical symptoms resulting from the R1448H mutation in contrast to the M1360V mutation. PMID- 12562903 TI - Sour taste stimuli evoke Ca2+ and pH responses in mouse taste cells. AB - Sour taste is elicited by acids. How taste cells transduce sour taste is controversial because acids (specifically protons) have diverse effects on cell membranes. Consequently, it is difficult to differentiate between events related to sour taste transduction per se and unrelated effects of protons. We have studied acid taste transduction in mouse taste buds using a lingual slice preparation where it is possible to measure changes in pH and [Ca2+]i simultaneously in taste cells. Focal application of citric acid or HCl to the apical tips of taste buds produced widespread acidification of the entire taste bud. Citric acid was effective at a pH of approximately 4, but HCl only at a pH of approximately 1.5. Despite acidification of the whole taste bud, only a select few taste cells exhibited Ca2+ responses. Acid-evoked Ca2+ responses were dose dependent in a range consistent with them being sour-taste responses. Cells exhibiting acid-evoked Ca2+ responses also responded to KCl depolarization. Acid evoked Ca2+ responses were blocked by Ba2+ (2 mM) and Cd2+ (500 microM), suggesting that acid responses are generated by Ca2+ influx through depolarization-gated Ca2+ channels. Removing extracellular Ca2+ reduced acid evoked Ca2+ responses, but depleting intracellular Ca2+ stores with thapsigargin had no effect, suggesting that acid taste responses are generated by an influx of extracellular Ca2+. Neither Cs+ (500 microM) nor amiloride (100 microM) affected acid-evoked Ca2+ responses, suggesting that neither hyperpolarization-activated cyclic nucleotide-gated cation (pacemaker) channels nor epithelial Na+ channels, respectively, transduce sour taste. Collectively, the results indicate that acids, especially weak acids, acidify the taste bud and evoke depolarization induced Ca2+ entry into a select subset of taste cells. The primary transducer protein(s) for sour taste remain undiscovered. PMID- 12562904 TI - Multiple conductance states of single Ca2+-activated Cl- channels in rabbit pulmonary artery smooth muscle cells. AB - Ca2+-activated Cl- channels contribute to agonist-evoked contraction and spontaneous activity in some smooth muscle preparations. Patch pipette techniques were used to study the properties of single Ca2+-activated Cl- channels in freshly dispersed rabbit pulmonary artery myocytes. In the cell-attached recording mode, two conductance states of 3.5 and 1.8 pS were recorded either spontaneously or in response to increasing [Ca2+]i. With inside-out patches, the 3.5 pS channel current predominated at 50 nM [Ca2+]i, but at 500 nM [Ca2+]i most channels opened to the 1.8 pS level and an additional 1.2 pS channel conductance was resolved. At 1 microM [Ca2+]i all of the Cl- channels opened either to the 1.8 pS or 1.2 pS level. In 0 [Ca2+]i, no channel activity was observed at -100 mV to +100 mV, but with 10-250 nM [Ca2+]i the total single channel open probability (NP(o)) increased with depolarisation. This voltage dependence was not seen at higher values of [Ca2+]i. The plot of NPo vs. [Ca2+]i yielded Ca2+ affinity constants of 8 and 250 nM and Hill slopes of 1.3 and 2.3 at +100 and -100 mV, respectively. The distribution of open times was fitted by two exponentials of about 5 and 30 ms, which were neither voltage nor Ca2+ dependent. Replacement of external Cl- by I- shifted the reversal potential by about -30 mV and lengthened the longer of the two mean open times without significant effects on other kinetic parameters. Based on these data, a model for the activation of Ca2+ activated Cl- channels is proposed. PMID- 12562905 TI - Spatial profiles of store-dependent calcium release in motoneurones of the nucleus hypoglossus from newborn mouse. AB - Hypoglossal motoneurones (HMN) are selectively damaged in both human amyotrophic lateral sclerosis (ALS) and corresponding mouse models of this neurodegenerative disease, a process which has been linked to their low endogenous Ca2+ buffering capacity and an exceptional vulnerability to Ca2+-mediated excitotoxic events. In this report, we investigated local Ca2+ profiles in low buffered HMNs by utilizing multiphoton microscopy, CCD imaging and patch clamp recordings in slice preparations. Bath application of caffeine induced highly localized Ca2+ release events, which displayed an initial peak followed by a slow 'shoulder' lasting several seconds. Peak amplitudes were paralleled by Ca2+-activated, apamin sensitive K+ currents (IKCa), demonstrating a functional link between Ca2+ stores and HMN excitability. The potential involvement of mitochondria was investigated by bath application of CCCP, which collapses the electrochemical potential across the inner mitochondrial membrane. CCCP reduced peak amplitudes of caffeine responses and consequently IKCa, indicating that functionally intact mitochondria were critical for store-dependent modulation of HMN excitability. Taken together, our results indicate localized Ca2+ release profiles in HMNs, where low buffering capacities enhance the role of Ca2+-regulating organelles as local determinants of [Ca2+]i. This might expose HMN to exceptional risks during pathophysiological organelle disruptions and other ALS-related, cellular disturbances. PMID- 12562906 TI - Enhanced G protein-dependent modulation of excitatory synaptic transmission in the cerebellum of the Ca2+ channel-mutant mouse, tottering. AB - Tottering, a mouse model for absence epilepsy and cerebellar ataxia, carries a mutation in the gene encoding class A (P/Q-type) Ca2+ channels, the dominant exocytotic Ca2+ channel at most synapses in the mammalian central nervous system. Comparing tottering to wild-type mice, we have studied glutamatergic transmission between parallel fibres and Purkinje cells in cerebellar slices. Results from biochemical assays and electrical field recordings demonstrate that glutamate release from parallel fibre terminals of the tottering mouse is controlled largely by class B Ca2+ channels (N-type), in contrast to the P/Q-channels that dominate release from wild-type terminals. Since N-channels, in a variety of assays, are more effectively inhibited by G proteins than are P/Q-channels, we tested whether synaptic transmission between parallel fibres and Purkinje cells in tottering mice was more susceptible to inhibitory modulation by G protein coupled receptors than in their wild-type counterparts. GABAB receptors and alpha2-adrenergic receptors (activated by bath application of transmitters) produced a three- to fivefold more potent inhibition of transmission in tottering than in wild-type synapses. This increased modulation is likely to be important for cerebellar transmission in vivo, since heterosynaptic depression, produced by activating GABAergic interneurones, greatly prolonged GABAB receptor-mediated presynaptic inhibition in tottering as compared to wild-type slices. We propose that this enhanced modulation shifts the balance of synaptic input to Purkinje cells in favour of inhibition, reducing Purkinje cell output from the cerebellum, and may contribute to the aberrant motor phenotype that is characteristic of this mutant animal. PMID- 12562907 TI - Inactivation of ICa-L is the major determinant of use-dependent facilitation in rat cardiomyocytes. AB - Two models have been proposed to explain facilitation of the L-type calcium current (ICa-L). A positive feedback model proposes that calcium released during a conditioning pulse (I1) facilitates the subsequent pulse (I2) via calmodulin/calmodulin kinase II (CaMKII) mechanisms. The negative feedback model proposes that the calcium release of each pulse feeds back on itself via calcium dependent inactivation. The relative physiological roles were evaluated in rat ventricular myocytes. Paired pulses (450 ms interpulse interval) elicited facilitation (I2 of 872 +/- 145 versus I1 of 777 +/- 132 pA, P < 0.01). Inactivation time (T0.37) was prolonged for I2 versus I1 (22 +/- 2 and 16 +/- 2 ms, P > 0.01). Evidence for the negative feedback mechanism includes: (a) ryanodine (0.3 mM ) eliminated facilitation, surprisingly by increasing the amplitude of I1 more than that of I2 (1039 +/- 216 and 977 +/- 186 pA) and eliminated the difference in T0.37 between I2 and I1 (33.1 +/- 4.5 versus 32.5 +/ 4.6 ms); (b) an outward I2, which does not trigger sarcoplasmic reticulum (SR) Ca2+ release, eliminated facilitation even when it was conditioned by an inward I1; (c) facilitation decayed as the I1-I2 interval lengthened (time constant (tau) = 16.9 +/- 1.4 s); (d) thapsigargin (0.1 microM ) slowed this decay (tau = 43.8 +/- 11.7 s) whereas isoproterenol accelerated it (tau = 5.6 +/- 1.4 s, P < 0.01) and T0.37 paralleled this decay; and (e) the magnitude of ICa-L was negatively correlated with the sodium-calcium exchange current (INa/Ca) elicited by the SR-Ca2+ release. In conclusion, Ca2+-dependent inactivation of ICa-L is the major mechanism underlying facilitation. PMID- 12562908 TI - Regulation of supply and demand for maternal nutrients in mammals by imprinted genes. AB - The placenta has evolved in eutherian mammals primarily to provide nutrients for the developing fetus. The genetic control of the regulation of supply and demand for maternal nutrients is not understood. In this review we argue that imprinted genes have central roles in controlling both the fetal demand for, and the placental supply of, maternal nutrients. Recent studies on Igf2 (insulin-like growth factor 2) knockout mouse models provide experimental support for this hypothesis. These show effects on placental transport capacity consistent with a role of IGF-II in modulating both the placental supply and fetal demand for nutrients. Imprinting of genes with such functions may have coevolved with the placenta and new evidence suggests that transporter proteins, as well as the regulators themselves, may also be imprinted. These data and hypotheses are important, as deregulation of supply and demand affects fetal growth and has long term consequences for health in mammals both in the neonatal period and, as a result of fetal programming, in adulthood. PMID- 12562909 TI - Local suppression of epileptiform activity by electrical stimulation in rat hippocampus in vitro. AB - High frequency electrical stimulation of deep brain structures (DBS) has been effective at controlling abnormal neuronal activity in Parkinson's patients and is now being applied for the treatment of pharmacologically intractable epilepsy. The mechanisms underlying the therapeutic effects of DBS are unknown. In particular, the effect of the electrical stimulation on neuronal firing remains poorly understood. Previous reports have showed that uniform electric fields with both AC (continuous sinusoidal) or DC waveforms could suppress epileptiform activity in vitro. In the present study, we tested the effects of monopolar electrode stimulation and low-duty cycle AC stimulation protocols, which more closely approximate those used clinically, on three in vitro epilepsy models. Continuous sinusoidal stimulation, 50 % duty-cycle sinusoidal stimulation, and low (1.68 %) duty-cycle pulsed stimulation (120 micros, 140 Hz) could completely suppress spontaneous low-Ca2+ epileptiform activity with average thresholds of 71.11 +/- 26.16 microA, 93.33 +/- 12.58 microA and 300 +/- 100 microA, respectively. Continuous sinusoidal stimulation could also completely suppress picrotoxin- and high-K+-induced epileptiform activity with either uniform or localized fields. The suppression generated by the monopolar electrode was localized to a region surrounding the stimulation electrode. Potassium concentration and transmembrane potential recordings showed that AC stimulation was associated with an increase in extracellular potassium concentration and neuronal depolarization block; AC stimulation efficacy was not orientation selective. In contrast, DC stimulation blocked activity by membrane hyperpolarization and was orientation-selective, but had a lower threshold for suppression. PMID- 12562910 TI - Enhanced excitability of myenteric AH neurones in the inflamed guinea-pig distal colon. AB - The electrical and synaptic properties of myenteric neurones in normal and inflamed guinea-pig distal colons were evaluated by intracellular microelectrode recording. Chronic inflammation was established 6 days following administration of trinitrobenzene sulfonic acid (TNBS). In S neurones, inflammation only altered synaptic inputs as the amplitude of fast excitatory postsynaptic potentials were significantly larger (31 +/- 2 mV compared to 20 +/- 1 mV) and they were more likely to receive slow excitatory synaptic input (85% compared to 55%). AH neurones displayed altered electrical properties in colitis compared to control tissues: they generated more action potentials during a maximal depolarising current pulse (7 +/- 1 compared to 1.6 +/- 0.2); they had a smaller after hyperpolarisation (9 +/- 2 mV s compared to 20 +/- 2 mV s); and they were more likely to receive fast excitatory synaptic input (74% compared to 17%), possess spontaneous activity (46% compared to 3%), and generate anodal break action potentials (58% compared to 19%). Although the resting membrane potential, input resistance and action potential characteristics were unaltered in AH neurones from inflamed tissues, they exhibited an enhanced Cs+-sensitive rectification of the current-voltage relationship. This suggests that the increase in excitability of AH neurones may involve a colitis-induced augmentation of the hyperpolarisation-activated cation current (Ih) in these cells. An increased excitability, selectively in AH neurones, suggests that the afferent limb of intrinsic motor reflexes is disrupted in the inflamed colon and this may contribute to dysmotility associated with inflammatory diseases. PMID- 12562911 TI - Molecular basis of the effect of potassium on heterologously expressed pacemaker (HCN) channels. AB - Hyperpolarization-activated cyclic-nucleotide-gated (HCN) channels modulate the firing rates of neuronal and cardiac pacemaker cells. HCN channels resemble voltage-gated K+ channels structurally, but much less is known about their structure-function correlation. Although modulation of K+ channel gating by external K+ is a well-known phenomenon, such a link has not been established for HCN channels. Here we examined the effects of external permeant (K+, Na+ and Li+) and non-permeant (NMG+) ions on HCN1 and HCN2 gating. Substituting 64 of 96 mM external K+ with Na+, Li+ or NMG+ positively shifted steady-state activation (approximately 13 mV), and preferentially slowed activation of HCN1. Mutating the pore variant C-terminal to the GYG motif in HCN1, A352, to the analogous conserved Asp in K+ channels or Arg in HCN2 produced a significant hyperpolarizing activation shift (by 5-15 mV), slowed gating kinetics (up to 6 fold), and abolished or attenuated gating responses to external K+. Whereas Na+, Li+ and NMG+ substitutions produced depolarizing activation shifts of HCN2 similar to those of HCN1, deactivation but not activation of HCN2 was exclusively decelerated. We conclude that gating and permeation of HCN channels are coupled, and that modulation of this 'pore-to-gate' coupling by external K+ is isoform specific. PMID- 12562912 TI - Effects of electrical stimulation and insulin on Na+-K+-ATPase ([3H]ouabain binding) in rat skeletal muscle. AB - Exercise has been reported to increase the Na+-K+-ATPase (Na+-K+ pump) alpha2 isoform in the plasma membrane 1.2- to 1.9-fold, purportedly reflecting Na+-K+ pump translocation from an undefined intracellular pool. We examined whether Na+ K+ pump stimulation, elicited by muscle contraction or insulin, increases the plasma membrane Na+-K+ pump content ([3H]ouabain binding) in muscles from young rats. Stimulation of isolated soleus muscle for 10 s at 120 Hz caused a rapid rise in intracellular Na+ content, followed by an 18-fold increase in the Na+ re extrusion rate (80 % of theoretical maximum). Muscles frozen immediately or 120 s after 10-120 s stimulation showed 10-22 % decrease in [3H]ouabain binding expressed per gram wet weight, but with no significant change expressed per gram dry weight. In soleus muscles from adult rats, [3H]ouabain binding was unaltered after 10 s stimulation at 120 Hz. Extensor digitorum longus (EDL) muscles stimulated for 10-60 s at 120 Hz showed no significant change in [3H]ouabain binding. Insulin (100 mU ml-1) decreased intracellular Na+ content by 27 % and increased 86Rb uptake by 23 % soleus muscles, but [3H]ouabain binding was unchanged. After stimulation for 30 s at 60 Hz soleus muscle showed a 30% decrease in intracellular Na+ content, demonstrating increased Na+-K+ pump activity, but [3H]ouabain binding measured 5 to 120 min after stimulation was unchanged. Stimulation of soleus or EDL muscles for 120-240 min at 1 Hz (continuously) or 10 Hz (intermittently) produced no change in [3H]ouabain binding per gram dry weight. In conclusion, the stimulating effects of electrical stimulation or insulin on active Na+, K+-transport in rat skeletal muscle could not be even partially accounted for by an acute increase in the content of functional Na+ -K+ pumps in the plasma membrane. PMID- 12562913 TI - Pulsatile control of the human masticatory muscles. AB - Spectral analysis of jaw acceleration confirmed that the human mandible 'trembles' at a peak frequency around 6 Hz when held in its rest position and at other stationary jaw openings. The 6 Hz tremor increased during very slow movements of the mandible, but other lower-frequency peaks became prominent during more rapid jaw movements. These lower-frequency peaks are likely to be the result of asymmetries in the underlying, voluntarily produced, 'saw-tooth' movements. In comparison, finger tremor at rest and during slow voluntary movements had a mean peak frequency of about 8 Hz: this frequency did not change during rhythmical finger flexion and extension movements, but the power of the tremor increased non-linearly with the speed of the movement. The resting jaw tremor was weakly coherent with the activity of the masseter and digastric muscles at the tremor frequency in about half the subjects, but was more strongly coherent during voluntary movements in all subjects. The masseter activity was at least 150 deg out of phase with the digastric activity at the tremor frequency (and at all frequencies from 2.5-15 Hz). The alternating pattern of activity in antagonistic muscles at rest and during slow voluntary movements supports the idea that the masticatory system is subject to pulsatile control in a manner analogous to that seen in the finger. PMID- 12562914 TI - ATP regulation of epithelial Cl- channels--new challenges? PMID- 12562915 TI - Troponin I in the murine myocardium: influence on length-dependent activation and interfilament spacing. AB - Cyclic AMP-dependent protein kinase (PKA) targets contractile proteins, troponin I (TnI) and myosin binding protein C (MyBP-C) in the heart and induces a decrease in myofilament Ca2+ sensitivity. Yet, the effect of sarcomere length (SL) change on Ca2+ sensitivity (length-dependent activation: LDA) following PKA-dependent phosphorylation is not clear. To clarify the role of PKA-dependent phosphorylation of TnI and MyBP-C on LDA in the heart, we examined LDA in skinned myocytes from a non-transgenic (NTG) and a transgenic murine model in which the native cardiac isoform (cTnI) was completely replaced by the slow skeletal isoform of TnI (ssTnI-TG) lacking the phosphorylation sites for PKA, while retaining PKA sites on MyBP-C. In NTG myocytes, PKA treatment decreased Ca2+ sensitivity at each SL, but enhanced the impact of SL change on Ca2+ sensitivity. Despite a greater sensitivity to Ca2+ and a reduction in LDA, neither Ca2+ responsiveness nor LDA was affected by PKA treatment in ssTnI-TG myocytes. To determine whether the above observations could be explained by the lateral separation between thick and thin filaments, as suggested by others, we measured interfilament spacing by X-ray diffraction as a function of SL in skinned cardiac trabeculae in the passive state from both NTG and ssTnI-TG models before and following treatment with PKA. Phosphorylation by PKA increased lattice spacing at every SL in NTG trabeculae. However, the relationship between SL and myofilament lattice spacing in ssTnI-TG was markedly shifted downward to an overall decreased myofilament lattice spacing following PKA treatment. We conclude: (1) PKA dependent phosphorylation enhances length-dependent activation in NTG hearts; (2) replacement of native TnI with ssTnI increases Ca2+ sensitivity of tension but reduces length-dependent activation; (3) MyBP-C phosphorylation by PKA does not alter calcium responsiveness and induces a decrease in myofilament lattice spacing at all sarcomere lengths and (4) length-dependent activation in the heart cannot be entirely explained by alterations in myofilament lattice spacing. PMID- 12562916 TI - Beta-oxidation of 5-hydroxydecanoate, a putative blocker of mitochondrial ATP sensitive potassium channels. AB - 5-Hydroxydecanoate (5-HD) inhibits ischaemic and pharmacological preconditioning of the heart. Since 5-HD is thought to inhibit specifically the putative mitochondrial ATP-sensitive K+ (KATP) channel, this channel has been inferred to be a mediator of preconditioning. However, it has recently been shown that 5-HD is a substrate for acyl-CoA synthetase, the mitochondrial enzyme which 'activates' fatty acids. Here, we tested whether activated 5-HD, 5 hydroxydecanoyl-CoA (5-HD-CoA), is a substrate for medium-chain acyl-CoA dehydrogenase (MCAD), the committed step of the mitochondrial beta-oxidation pathway. Using a molecular model, we predicted that the hydroxyl group on the acyl tail of 5-HD-CoA would not sterically hinder the active site of MCAD. Indeed, we found that 5-HD-CoA was a substrate for purified human liver MCAD with a Km of 12.8 +/- 0.6 microM and a kcat of 14.1 s-1. For comparison, with decanoyl CoA (Km approximately 3 microM) as substrate, kcat was 6.4 s-1. 5-HD-CoA was also a substrate for purified pig kidney MCAD. We next tested whether the reaction product, 5-hydroxydecenoyl-CoA (5-HD-enoyl-CoA), was a substrate for enoyl-CoA hydratase, the second enzyme of the beta-oxidation pathway. Similar to decenoyl CoA, purified 5-HD-enoyl-CoA was also a substrate for the hydratase reaction. In conclusion, we have shown that 5-HD is metabolised at least as far as the third enzyme of the beta-oxidation pathway. Our results open the possibility that beta oxidation of 5-HD or metabolic intermediates of 5-HD may be responsible for the inhibitory effects of 5-HD on preconditioning of the heart. PMID- 12562917 TI - Regulation of an inactivating potassium current (IA) by the extracellular matrix protein vitronectin in embryonic mouse hippocampal neurones. AB - Integrins are a class of intrinsic membrane receptors for extracellular matrix ligands. In the central nervous system, integrins and their ligands influence neuronal growth and synaptic function, but relatively little is known about their potential to regulate intrinsic excitability. To explore this area, we examined the effects of matrix components on potassium currents in developing mouse hippocampal neurones, using electrophysiological and immunochemical approaches. We tested the effects of three integrin ligands present in the hippocampus, fibronectin, laminin and vitronectin, on electrogenesis in late embryonic hippocampal pyramidal neurones. Explants cultured in serum-free medium were exposed to ligands (fibronectin at 3 microg ml-1, laminin at 5 microg ml-1, vitronectin at 10 microg ml-1) for 3-4 days, and voltage-gated potassium currents were recorded from presumptive CA3 pyramidal neurones. Of the three matrix components, only vitronectin affected potassium currents, selectively increasing the amplitude of the inactivating potassium current (IA, or A-current) by about 75 % over control levels, and its density (current per unit area) by about 40 % (measured after 3 day exposures from embryonic day 15.5). Other potassium currents were spared, except to the extent that membrane area was increased. The actions of vitronectin were sensitive to RGD (Arg-Gly-Asp)-sequence-containing peptide, indicating the involvement of integrins as vitronectin receptors. The kinetic properties of IA, including the voltage-dependence of activation and inactivation, inactivation rate and the rate of recovery from inactivation, were minimally affected by vitronectin and were consistent with enhanced functional expression of Kv4-family subunits. Analyses of Kv4.2 and Kv1.4 immunoreactivity also suggested a preferential increase in Kv4.2 levels, with lesser effects on Kv1.4 levels. These results indicate that vitronectin can selectively regulate IA, and together with other observations suggest that modulation of neuronal excitability by integrins and their ligands occurs commonly. PMID- 12562918 TI - Mitogen-activated protein kinase (MAPK) pathway activation: effects of age and acute exercise on human skeletal muscle. AB - The purpose of this investigation was to examine the activation (phosphorylation) and total protein content of MAPK signalling cascade proteins (ERK 1/2, p90RSK, Mnk 1, eIF4E, p38 MAPK, JNK/SAPK, MKP 1) at rest and following exercise, in sedentary young and old men. Eight young (22 +/- 1 years; YM) and eight old (79 +/- 3 years; OM) men underwent a resting muscle biopsy of the vastus lateralis; they then performed a knee extensor resistance exercise session (29 contractions at approximately 70 % of max), followed by a post-exercise biopsy. Western immunoblot analysis demonstrated that the OM had higher resting phosphorylation of ERK 1/2, p90RSK, Mnk 1, p38 MAPK and JNK/SAPK proteins versus YM (P < 0.05). The resistance exercise bout caused an increase in phosphorylation of the ERK 1/2, p90RSK and Mnk 1 proteins (P < 0.05) in the YM. Conversely, the OM had a decrease in ERK 1/2, p90RSK, Mnk 1, p38 MAPK and JNK/SAPK phosphorylation (P < 0.05) after the exercise bout. Neither group showed a change in eIF4E phosphorylation. The total amount of protein expression of the MAPK signalling proteins was not different between the YM and OM, except that there was a higher (P < 0.05) MKP 1 protein content in the OM. This investigation is the first to provide evidence that MAPK proteins are differentially activated at rest and in response to a bout of resistance exercise in skeletal muscle of young and old men. These findings may have implications for other processes (e.g. transcription and translation) involved in skeletal muscle type and growth, when examining the changes occurring with ageing muscle before and after resistance exercise/training. PMID- 12562919 TI - Lifetime consequences of abnormal fetal pancreatic development. AB - There is ample evidence that an adverse intrauterine environment has harmful consequences for health in later life. Maternal diabetes and experimentally induced hyperglycaemia result in asymmetric overgrowth, which is associated with an increased insulin secretion and hyperplasia of the insulin-producing B-cells in the fetuses. In adult life, a reduced insulin secretion is found. In contrast, intrauterine growth restriction is associated with low insulin secretion and a delayed development of the insulin-producing B-cells. These perinatal alterations may induce a deficient adaptation of the endocrine pancreas and insulin resistance in later life. Intrauterine growth restriction in human pregnancy is mainly due to a reduced uteroplacental blood flow or to maternal undernutrition or malnutrition. However, intrauterine growth restriction can be present in severe diabetes complicated by vasculopathy and nephropathy. In animal models, intrauterine growth retardation can be obtained through pharmacological (streptozotocin), dietary (semi-starvation, low protein diet) or surgical (intrauterine artery ligation) manipulation of the maternal animal. The endocrine pancreas and more specifically the insulin-producing B-cells play an important role in the adaptation to an adverse intrauterine milieu and the consequences in later life. The long-term consequences of an unfavourable intrauterine environment are of major importance worldwide. Concerted efforts are needed to explore how these long-term effects can be prevented. This review will consist of two parts. In the first part, we discuss the long-term consequences in relation to the development of the fetal endocrine pancreas and fetal growth in the human; in the second part, we focus on animal models with disturbed fetal and pancreatic development and the consequences for later life. PMID- 12562920 TI - Cl- flux through a non-selective, stretch-sensitive conductance influences the outer hair cell motor of the guinea-pig. AB - Outer hair cells underlie high frequency cochlear amplification in mammals. Fast somatic motility can be driven by voltage-dependent conformational changes in the motor protein, prestin, which resides exclusively within lateral plasma membrane of the cell. Yet, how a voltage-driven motor could contribute to high frequency amplification, despite the low-pass membrane filter of the cell, remains an enigma. The recent identification of prestin's Cl- sensitivity revealed an alternative mechanism in which intracellular Cl- fluctuations near prestin could influence the motor. We report the existence of a stretch-sensitive conductance within the lateral membrane that passes anions and cations and is gated at acoustic rates. The resultant intracellular Cl- oscillations near prestin may drive motor protein transitions, as evidenced by pronounced shifts in prestin's state-probability function along the voltage axis. The sensitivity of prestin's state probability to intracellular Cl- levels betokens a more complicated role for Cl- than a simple extrinsic voltage sensor. Instead, we suggest an allosteric modulation of prestin by Cl- and other anions. Finally, we hypothesize that prestin sensitivity to anion flux through the mechanically activated lateral membrane can provide a driving force that circumvents the membrane's low-pass filter, thus permitting amplification at high acoustic frequencies. PMID- 12562921 TI - Mouse muscle denervation increases expression of an alpha7 nicotinic receptor with unusual pharmacology. AB - Neuronal nicotinic alpha7 subunits have been found in chick and rat skeletal muscle during development and denervation. In the present study, reverse transcriptase-polymerase chain reaction was used to detect alpha7 subunit mRNA in denervated mouse muscle. To determine whether the alpha7 subunit forms functional nicotinic acetylcholine receptors (nAChRs) in muscle, choline was used to induce a membrane depolarization because choline has been considered a specific agonist of alpha7-containing (alpha7*) nAChRs. We found, however, that choline (3-10 mM) also weakly activates muscle nAChRs. After inhibiting muscle nAChRs with a specific muscle nAChR inhibitor, alpha-conotoxin GI (alphaCTxGI), choline was used to activate the alpha7* nAChRs on muscle selectively. Four weeks after denervation, rapid application of choline (10 mM) elicited a substantial depolarization in the presence of alphaCTxGI (0.1 microM). This component of the depolarization was never present in denervated muscles obtained from mutant mice lacking the alpha7 subunit (i.e. alpha7-null mice). The depolarization component that is resistant to alphaCTxGI was antagonized by pancuronium (3-10 microM) and by a 4-oxystilbene derivative (F3, 0.1-0.5 microM) at concentrations considered highly specific for alpha7* nAChRs. Another selective alpha7 antagonist, methyllycaconitine (0.05-5 microM), did not strongly inhibit this choline-induced depolarization. Furthermore, the choline-sensitive nAChRs showed little desensitization over 10 s of application with choline (10-30 mM). These results indicate that functional alpha7* nAChRs are significantly present on denervated muscle, and that these receptors display unusual functional and pharmacological characteristics. PMID- 12562922 TI - Metabolic regulation of Ca2+ release in permeabilized mammalian skeletal muscle fibres. AB - In the present study, the link between cellular metabolism and Ca2+ signalling was investigated in permeabilized mammalian skeletal muscle. Spontaneous events of Ca2+ release from the sarcoplasmic reticulum were detected with fluo-3 and confocal scanning microscopy. Mitochondrial functions were monitored by measuring local changes in mitochondrial membrane potential (with the potential-sensitive dye tetramethylrhodamine ethyl ester) and in mitochondrial [Ca2+] (with the Ca2+ indicator mag-rhod-2). Digital fluorescence imaging microscopy was used to quantify changes in the mitochondrial autofluorescence of NAD(P)H. When fibres were immersed in a solution without mitochondrial substrates, Ca2+ release events were readily observed. The addition of L-glutamate or pyruvate reversibly decreased the frequency of Ca2+ release events and increased mitochondrial membrane potential and NAD(P)H production. Application of various mitochondrial inhibitors led to the loss of mitochondrial [Ca2+] and promoted spontaneous Ca2+ release from the sarcoplasmic reticulum. In many cases, the increase in the frequency of Ca2+ release events was not accompanied by a rise in global [Ca2+]i. Our results suggest that mitochondria exert a negative control over Ca2+ signalling in skeletal muscle by buffering Ca2+ near Ca2+ release channels. PMID- 12562923 TI - An inwardly rectifying K+ channel in bovine parotid acinar cells: possible involvement of Kir2.1. AB - Using electrophysiological and molecular techniques, we investigated the molecular nature of an inwardly rectifying K+ channel in bovine parotid acinar (BPA) cells and examined its role in setting resting membrane potential. In whole cell recordings from freshly isolated BPA cells, a predominant current was a K+ current rectified strongly in the inward direction. An inward conductance of the inwardly rectifying K+ (Kir) current was proportional to [K+]o(0.57). The selectivity sequence based on permeability ratios was K+ (1.00) > Rb+ (0.63) >> Li+ (0.04) = Na+ (0.02) and the sequence based on conductance ratios was K+ (1.00) >> Rb+ (0.03) = Li+ (0.03) = Na+ (0.02). The current was blocked by extracellular Ba2+ and Cs+ in a voltage- and a concentration-dependent manner, with a Kd at 0 mV of 11.6 microM and 121 mM, respectively. Cell-attached patch measurements identified 27 pS K+ channels as being the most likely to mediate whole-cell Kir currents. Addition of Ba2+ (100 microM) to the bathing solution reversibly depolarized the resting membrane potential in intact unstimulated cells. RT-PCR of RNA from bovine parotid cells revealed transcripts of bovine Kir2.1 (bKir2.1). HEK293 cells stably expressing bKir2.1 cloned from bovine parotid exhibited whole-cell and single channel Kir currents, of which electrophysiological characteristics were quantitatively similar to those of native Kir currents. Immunohistochemical studies showed a bKir2.1 immunoreactivity in BPA cells. Collectively, these results suggest that Kir2.1 may mediate native Kir currents responsible for setting resting membrane potential in BPA cells and might be, at least in part, involved in spontaneous secretion in ruminant parotid glands. PMID- 12562924 TI - The smooth muscle myosin seven amino acid heavy chain insert's kinetic role in the crossbridge cycle for mouse bladder. AB - The seven amino acid insert in the smooth muscle myosin heavy chain is thought to regulate the kinetics of contraction, contributing to the differences between fast and slow smooth muscle. The effects of this insert on force and stiffness were determined in bladder tissue of a transgenic mouse line expressing the insert SMB at one of three levels: an SMB wild type (+/+), an SMA homozygous type (-/-) and a heterozygous type (+/-). For skinned muscle, an increase in MgADP or inorganic phosphate (Pi) should shift the distribution of crossbridges in the actomyosin ATPase (AMATPase) to increase the relative population of the crossbridge state prior to ADP release and Pi release, respectively. Exogenous ADP increased force and stiffness in a manner consistent with increasing the Ca2+ concentration in both the +/+ and +/- mouse types. However, the -/- type showed a significantly greater increase in force than in stiffness suggesting that immediately prior to ADP release, the AMATPase either has an additional force producing isomerization state or a slower ADP dissociation rate for the -/- type compared to the +/+ or +/- types. Exogenous Pi led to a significantly greater decrease in stiffness than in force for all three mouse types suggesting that there is a force producing state prior to Pi release. In addition, the increase in Pi showed similar changes in the +/+ and -/- types whereas in the +/- type the decreases in both force and stiffness were greater than the other two mouse types indicating that the insert can affect the cooperativity between myosin heads. In conclusion, the seven amino acid insert modulates the kinetics and/or states of the AMATPase, which could lead to differences in the kinetics of contraction between fast and slow smooth muscle. PMID- 12562925 TI - Ryanodine stores and calcium regulation in the inner segments of salamander rods and cones. AB - Despite the prominent role played by intracellular Ca2+ stores in the regulation of neuronal Ca2+ homeostasis and in invertebrate photoreception, little is known about their contribution to the control of free Ca2+ concentration ([Ca2+]i) in the inner segments of vertebrate photoreceptors. Previously, caffeine-sensitive intracellular Ca2+ stores were shown to play a role in regulating glutamate release from photoreceptors. To understand the properties of these intracellular stores better we used pharmacological approaches that alter the dynamics of storage and release of Ca2+ from intracellular compartments. Caffeine evoked readily discernible changes in [Ca2+]i in the inner segments of rods, but not cones. Caffeine-evoked Ca2+ responses in cone inner segments were unmasked in the presence of inhibitors of the plasma membrane Ca2+ ATPases (PMCAs) and mitochondrial Ca2+ sequestration. Caffeine-evoked responses were blocked by ryanodine, a selective blocker of Ca2+ release and by cyclopiazonic acid, a blocker of Ca2+ sequestration into the endoplasmic reticulum. These two inhibitors also substantially reduced the amplitude of depolarization-evoked [Ca2+]i increases, providing evidence for Ca2+-induced Ca2+ release (CICR) in rods and cones. The magnitude and kinetics of caffeine-evoked Ca2+ elevation depended on the basal [Ca2+]i, PMCA activity and on mitochondrial function. These results reveal an intimate interaction between the endoplasmic reticulum, voltage gated Ca2+ channels, PMCAs and mitochondrial Ca2+ stores in photoreceptor inner segments, and suggest a role for CICR in the regulation of synaptic transmission. PMID- 12562926 TI - Amyloid beta(1-42) peptide alters the gating of human and mouse alpha bungarotoxin-sensitive nicotinic receptors. AB - The beta-amyloid(1-42) peptide (Abeta(1-42)), a major constituent of the Alzheimer's disease amyloid plaque, specifically binds to the neuronal alpha bungarotoxin (alpha-BuTx)-sensitive alpha7 nicotinic acetylcholine receptor (alpha7 nAChR). Accordingly, Abeta1-42 interferes with the function of alpha7 nAChRs in chick and rodent neurons. To gain insights into the human disease, we studied the action of Abeta(1-42) on human alpha7 nAChRs expressed in Xenopus oocytes. In voltage-clamped oocytes expressing the wild-type receptor, Abeta(1 42) blocked ACh-evoked currents. The block was non-competitive, required over 100 s to develop and was partially reversible. In oocytes expressing the mutant L248T receptor, Abeta(1-42) activated methyllycaconitine-sensitive currents in a dose dependent manner. Peptide-evoked unitary events, recorded in outside-out patches, showed single-channel conductances and open duration comparable to ACh-evoked events. Abeta(1-42) had no effect on the currents evoked by glutamate, GABA or glycine in oocytes expressing human or mouse receptors for these transmitters. Muscle nAChRs are also alpha-BuTx-sensitive and we therefore investigated whether they respond to Abeta(1-42). In human kidney BOSC 23 cells expressing the fetal or adult mouse muscle nAChRs, Abeta(1-42) blocked ACh-evoked whole-cell currents, accelerating their decay. Outside-out single-channel recordings showed that the block was due to a reduced channel open probability and enhanced block upon ACh application. We also report that the inverse peptide Abeta(42-1), but not Abeta(40-1), partially mimicked the effects of the physiological Abeta(1-42) peptide. Possible implications for degenerative neuronal and muscular diseases are discussed. PMID- 12562927 TI - Calcium channel heterogeneity in canine left ventricular myocytes. AB - Regional variations in the electrophysiological properties of myocytes across the left ventricular wall play an important role in both the normal physiology of the heart and the genesis of arrhythmias. To investigate the possible contributions of calcium channels to transmural electrical heterogeneity, whole-cell patch clamp recordings were made from isolated canine epicardial and endocardial left ventricular myocytes. Two major differences in Ca2+ channel properties were found between epi- and endocardial cells. First, the L-type Ca2+ current was larger in endocardial than in epicardial myocytes. The average peak current density at +10 mV in endocardial myocytes was 3.4 +/- 0.2 pA pF-1, and was 45 % higher than that in epicardium (2.3 +/- 0.1 pA pF-1). The kinetic properties of the L-type current in epi- and endocardial cells were not significantly different. Second, a low threshold, rapidly activating and inactivating Ca2+ current that resembled the T type current was present in all endocardial myocytes but was small or absent in epicardial myocytes. This T-like current had an average peak density of 0.5 pA pF 1 at -40 mV in endocardial cells. In most endocardial cells the T-like Ca2+ current comprised two components: a Ni2+-sensitive T-type current and a tetrodotoxin-sensitive Ca2+ current. We conclude that there are considerable regional variations in the density and properties of Ca2+ channels across the canine left ventricular wall. These variations may contribute to the overall transmural electrical heterogeneity. PMID- 12562928 TI - An antisense oligonucleotide against H1 inhibits the classical sodium current but not ICa(TTX) in rat ventricular cells. AB - ICa(TTX) is a sodium current component, functionally distinct from the main body of sodium current, seen in cardiac and other cells. To determine if ICa(TTX) channels are a separate isoform from the classical cardiac sodium channels, we exposed rat ventricular cells in primary culture to an antisense oligonucleotide (AON) directed against rH1 (rNav1.5): 5'-CTCCTCATACCCTCT-3'. The homologous human sequence has been identified (and confirmed by us on HEK 293 cells) as effective against hH1 expressed heterologously. Scrambled sequence (5'-CCCCCCTTATCTACT-3') controls were also included. The AON (10 microM; day 2 of exposure) reduced the classical sodium current by 69.6 % compared to untreated and 60.8 % compared to scrambled sequence (10 microM; day 2 of exposure) controls (mean +/- S.E.M. maximum peak inward current density of -8.23 +/- 0.60 pA pF-1, 18 cells, for untreated; -6.37 +/- 0.79 pA pF-1, 16 cells, for scrambled sequence; and -2.50 +/ 0.31 pA pF-1, 18 cells, for AON-treated cells). The two control groups are not significantly different from each other, but are both significantly different from the AON-treated group (P < 0.001). The inhibition was specific for sodium channels, with no significant AON effect on the L-type calcium current. This confirms that H1 generates the classical cardiac sodium current. This same AON at the same concentration and time of exposure had no significant effect on ICa(TTX) (mean of -4.72 +/- 0.55, 15 cells; -5.47 +/- 0.53, 13 cells; and -5.04 +/- 0.63 pA pF-1, 15 cells, for untreated controls, scrambled controls and AON treated, respectively). Hence, ICa(TTX), which is functionally distinct from the classical cardiac sodium current, is encoded by a distinct gene. PMID- 12562929 TI - Effects of oxidation and cytosolic redox conditions on excitation-contraction coupling in rat skeletal muscle. AB - In this study the effects of oxidation and reduction on various steps in the excitation-contraction (E-C) coupling sequence was examined in mammalian skeletal muscle. In mechanically skinned fast-twitch fibres, electric field stimulation was used to generate action potentials in the sealed transverse-tubular (T-) system, thereby eliciting twitch responses, which are a sensitive measure of Ca2+ release. Treatment of fibres with the oxidant H2O2 (200 microM and 10 mM) for 2-5 min markedly potentiated caffeine-induced Ca2+ release and the force response to partial depolarisation of the T-system (by solution substitution). Importantly, such H2O2 treatment had no effect at all on any aspect of the twitch response (peak amplitude, rate of rise, decay rate constant and half-width), except in cases where it interfered with the T-system potential or voltage-sensor activation, resulting in a reduction or abolition of the twitch response. Exposure to strong thiol reductants, dithiothreitol (DTT, 10 mM) and reduced glutathione (GSH, 5 mM), did not affect the twitch response over 5 min, nor did varying the glutathione ratio (reduced to oxidised glutathione) from the level present endogenously in the cytosol of a rested fibre (30:1) to the comparatively oxidised level of 3:1. In fibres that had been oxidised by H2O2 (10 mM) (or by 2,2'-dithiodipyridine, 100 microM), exposure to GSH (5 mM) caused potentiation of twitch force (by approximately 20 % for H2O2); this effect was due to the increase in the Ca2+ sensitivity of the contractile apparatus that occurs under such circumstances and was fully reversed by subsequent exposure to 10 mM DTT. We conclude that: (a) the redox potential across the sarcomplamsic reticulum has no noticeable direct effect on normal E-C coupling in mammalian skeletal muscle, (b) oxidising the Ca2+-release channels and greatly increasing their sensitivity to Ca2+-induced Ca2+ release does not alter the amount of Ca2+ released by an action potential and (c) oxidation potentiates twitches by a GSH-mediated increase in the Ca2+ sensitivity of the contractile apparatus. PMID- 12562930 TI - Crucial role of sodium channel fast inactivation in muscle fibre inexcitability in a rat model of critical illness myopathy. AB - Critical illness myopathy is an acquired disorder in which skeletal muscle becomes electrically inexcitable. We previously demonstrated that inactivation of Na+ channels contributes to inexcitability of affected fibres in an animal model of critical illness myopathy in which denervated rat skeletal muscle is treated with corticosteroids (steroid denervated; SD). Our previous work, however, did not address the relative importance of membrane depolarization versus a shift in the voltage dependence of fast inactivation in causing inexcitability. It also remained unknown whether changes in the voltage dependence of activation or slow inactivation play a role in inexcitability. In the current study we found that a hyperpolarizing shift in the voltage dependence of fast inactivation of Na+ channels is the principal factor underlying inexcitability in SD fibres. Although depolarization tends to decrease excitability, it is insufficient to account for inexcitability in SD fibres since many normal and denervated fibres retain normal excitability when depolarized to the same resting potentials as affected SD fibres. Changes in the voltage dependence of activation and slow inactivation of Na+ channels were also observed in SD fibres; however, the changes appear to increase rather than decrease excitability. These results highlight the importance of the change in fast inactivation in causing inexcitability of SD fibres. PMID- 12562931 TI - Role for the cholecystokinin-A receptor in fever: a study of a mutant rat strain and a pharmacological analysis. AB - The involvement of the cholecystokinin (CCK)-A receptor in fever was studied. The polyphasic febrile responses to lipopolysaccharide (LPS; 10 microg kg-1, I.V.) were compared between wild-type Long-Evans (LE) rats and the CCK-A-receptor deficient Otsuka LE Tokushima Fatty (OLETF) rats. The response of the wild-type rats was biphasic, which is typical for LE rats. Phases 1 and 2 of the response of the OLETF rats were similar to those of the LE rats, but the OLETF rats also developed a robust phase 3. This late enhancement of the febrile response could reflect either the absence of the A receptor per se or a secondary trait of the mutant strain. To distinguish between these possibilities, we conducted a pharmacological analysis. We studied whether the normally low phase 3 of LE rats can be enhanced by a CCK-A-receptor antagonist, sodium lorglumide (4.3 microg kg 1 min-1, 120 min, I.V.), and whether the normally high phase 3 of Wistar rats can be attenuated by a CCK-A receptor agonist, sulphated CCK-8 (up to 0.17 microg kg 1 min-1, 120 min, I.V.). The dose of sodium lorglumide used was sufficient to increase food intake (to block satiety), but it did not affect the fever response. In both febrile and afebrile rats, CCK-8 induced dose-dependent skin vasodilatation and decreased body temperature, but it failed to produce any effects specific for phase 3. We conclude that the exaggeration of phase 3 in OLETF rats reflects a secondary trait of this strain and not the lack of the CCK A receptor per se. None of the three known phases of the febrile response of rats to LPS requires the CCK-A receptor. PMID- 12562932 TI - Ketamine increases human motor cortex excitability to transcranial magnetic stimulation. AB - Subanaesthetic doses of the N-methyl-D-aspartate (NMDA) antagonist ketamine have been shown to determine a dual modulating effect on glutamatergic transmission in experimental animals, blocking NMDA receptor activity and enhancing non-NMDA transmission through an increase in the release of endogenous glutamate. Little is known about the effects of ketamine on the excitability of the human central nervous system. The effects of subanaesthetic, graded incremental doses of ketamine (0.01, 0.02 and 0.04 mg kg-1 min-1, I.V.) on the excitability of cortical networks of the human motor cortex were examined with a range of transcranial magnetic and electric stimulation protocols in seven normal subjects. Administration of ketamine at increasing doses produced a progressive reduction in the mean resting motor threshold (RMT) (F(3, 18) = 22.33, P < 0.001) and active motor threshold (AMT) (F(3, 18) = 12.17, P < 0.001). Before ketamine administration, mean RMT +/- S.D. was 49 +/- 3.3 % of maximum stimulator output and at the highest infusion level it was 42.6 +/- 2.6 % (P < 0.001). Before ketamine administration, AMT +/- S.D. was 38 +/- 3.3 % of maximum stimulator output and at the highest infusion level it was 33 +/- 4.4 % (P < 0.002). Ketamine also led to an increase in the amplitude of EMG responses evoked by magnetic stimulation at rest; this increase was a function of ketamine dosage (F(3, 18) = 5.29, P = 0.009). In contrast to responses evoked by magnetic stimulation, responses evoked by electric stimulation were not modified by ketamine. The differential effect of ketamine on responses evoked by magnetic and electric stimulation demonstrates that subanaesthetic doses of ketamine enhance the recruitment of excitatory cortical networks in motor cortex. Transcranial magnetic stimulation produces a high-frequency repetitive discharge of pyramidal neurones and for this reason probably depends mostly on short-lasting AMPA transmission. An increase in this transmission might facilitate the repetitive discharge of pyramidal cells after transcranial magnetic stimulation which, in turn, results in larger motor responses and lower thresholds. We suggest that the enhancement of human motor cortex excitability to transcranial magnetic stimulation is the effect of an increase in glutamatergic transmission at non NMDA receptors similar to that described in experimental studies. PMID- 12562933 TI - Rod and cone contributions to the a-wave of the electroretinogram of the macaque. AB - The electroretinogram (ERG) of anaesthetised dark-adapted macaque monkeys was recorded in response to ganzfeld stimulation and rod- and cone-driven receptoral and postreceptoral components were separated and modelled. The test stimuli were brief (< 4.1 ms) flashes. The cone-driven component was isolated by delivering the stimulus shortly after a rod-saturating background had been extinguished. The rod-driven component was derived by subtracting the cone-driven component from the mixed rod-cone ERG. The initial part of the leading edge of the rod-driven a wave scaled linearly with stimulus energy when energy was sufficiently low and, for times less than about 12 ms after the stimulus, it was well described by a linear model incorporating a distributed delay and three cascaded low-pass filter elements. Addition of a simple static saturating non-linearity with a characteristic intermediate between a hyperbolic and an exponential function was sufficient to extend application of the model to most of the leading edge of the saturated responses to high energy stimuli. It was not necessary to assume involvement of any other non-linearity or that any significant low-pass filter followed the non-linear stage of the model. A negative inner-retinal component contributed to the later part of the rod-driven a-wave. After suppressing this component by blocking ionotropic glutamate receptors, the entire a-wave up to the time of the first zero-crossing scaled with stimulus energy and was well described by summing the response of the rod model with that of a model describing the leading edge of the rod-bipolar cell response. The negative inner retinal component essentially cancelled the early part of the rod-bipolar cell component and, for stimuli of moderate energy, made it appear that the photoreceptor current was the only significant component of the leading edge of the a-wave. The leading edge of the cone-driven a-wave included a slow phase that continued up to the peak, and was reduced in amplitude either by a rod suppressing background or by the glutamate analogue, cis-piperidine-2,3 dicarboxylic acid (PDA). Thus the slow phase represents a postreceptoral component present in addition to a fast component of the a-wave generated by the cones themselves. At high stimulus energies, it appeared less than 5 ms after the stimulus. The leading edge of the cone-driven a-wave was adequately modelled as the sum of the output of a cone photoreceptor model similar to that for rods and a postreceptoral signal obtained by a single integration of the cone output. In addition, the output of the static non-linear stage in the cone model was subject to a low-pass filter with a time constant of no more than 1 ms. In conclusion, postreceptoral components must be taken into account when interpreting the leading edge of the rod- and cone-driven a-waves of the dark-adapted ERG. PMID- 12562934 TI - Variation in large-conductance, calcium-activated potassium channels from hair cells along the chicken basilar papilla. AB - The mechanism for electrical tuning in non-mammalian hair cells rests within the widely diverse kinetics of functionally distinct, large-conductance potassium channels (BK), thought to result from alternative splicing of the pore-forming alpha subunit and variable co-expression with an accessory beta subunit. Inside out patches from hair cells along the chicken basilar papilla revealed 'tonotopic' gradations in calcium sensitivity and deactivation kinetics. The resonant frequency for the hair cell from which the patch was taken was estimated from deactivation rates, and this frequency reasonably matched that predicted from the originating cell's tonotopic location. The rates of deactivation for native BK channels were much faster than rates reported for cloned chicken BK channels including both alpha and beta subunits. This result was surprising since patches were pulled from hair cells in the apical half of the papilla where beta subunits are most highly expressed. Heterogeneity in the properties of native chicken BK channels implies a high degree of molecular variation and hinders our ability to identify those molecular constituents. PMID- 12562936 TI - Response of human jaw muscles to axial stimulation of the incisor. AB - The role of periodontal mechanoreceptors (PMRs) in the reflex control of the jaw muscles has thus far been mainly derived from animal studies. To date, the work that has been done on humans has been limited and confined to orthogonal stimulation of the labial surface of the tooth. The purpose of this study was to investigate the response of the masseter and digastric muscles in humans to controlled axial stimulation of the upper left central incisor, both before and during a local anaesthetic block of the PMRs. Ten neurologically normal young adult females were tested, each on two separate occasions to confirm the reproducibility of the results. It was found that the reflex response in the masseter was modulated by the rate of rise of the stimulus used and, to a lesser degree, the level of background muscle activity. There was little detectable change in the activity of the digastric muscle under the tested conditions and what was found could be attributed to cross-talk with the masseter. The reflex responses obtained were significantly different between subjects; however retesting the same subject on a different occasion yielded similar results. The results indicate that the most common response of the masseter muscle to brisk axial stimulation of the incisor is a reflex inhibition at 20 ms, followed by a late excitation at 44 ms. However, it is possible that this late excitation could be due to delayed action potentials and hence be artefactual. As the application of a local anaesthetic block removed or significantly reduced both of these responses, it was concluded that they originated from the PMRs. Unlike during orthogonal stimulation, slowly rising stimuli did not produce any excitatory reflex activity. This indicated a difference in jaw reflexes to forces applied in different directions, possibly due to the activation of different receptor types when stimulating the tooth in either the orthogonal or axial directions. PMID- 12562935 TI - Release of ATP induced by hypertonic solutions in Xenopus oocytes. AB - ATP mediates intercellular communication. Mechanical stress and changes in cell volume induce ATP release from various cell types, both secretory and non secretory. In the present study, we stressed Xenopus oocytes with a hypertonic solution enriched in mannitol (300 mM). We measured simultaneously ATP release and ionic currents from a single oocyte. A decrease in cell volume, the activation of an inward current and ATP release were coincident. We found two components of ATP release: the first was associated with granule or vesicle exocytosis, because it was inhibited by tetanus neurotoxin, and the second was related to the inward current. A single exponential described the correlation between ATP release and the hypertonic-activated current. Gadolinium ions, which block mechanically activated ionic channels, inhibited the ATP release and the inward current but did not affect the decrease in volume. Oocytes expressing CFTR (cystic fibrosis transmembrane regulator) released ATP under hypertonic shock, but ATP release was significantly inhibited in the first component: that related to granule exocytosis. Since the ATP measured is the balance between ATP release and ATP degradation by ecto-enzymes, we measured the nucleoside triphosphate diphosphohydrolase (NTPDase) activity of the oocyte surface during osmotic stress, as the calcium-dependent hydrolysis of ATP, which was inhibited by more than 50 % in hypertonic conditions. The best-characterized membrane protein showing NTPDase activity is CD39. Oocytes injected with an antisense oligonucleotide complementary to CD39 mRNA released less ATP and showed a lower amplitude in the inward current than those oocytes injected with water. PMID- 12562937 TI - Effect of atropine on the biomechanical properties of the oesophageal wall in humans. AB - Recently, we reported a novel ultrasound technique to assess biomechanical properties of the oesophagus in human subjects. In the present study, we use the technique, in combination with atropine, to determine the active and passive biomechanical properties of the oesophagus in normal healthy humans. A manometric catheter equipped with a high-compliance bag and a high-frequency intraluminal ultrasonography probe was used to record pressure and oesophageal geometry. Oesophageal distensions with either isovolumic (5-20 ml water) or with isobaric (10-60 mmHg) technique were performed. Intra-bag pressure and ultrasound images of the oesophagus were recorded simultaneously. Following injection of atropine (15 microg kg-1, I.V.), the oesophageal distensions were repeated. The oesophageal wall compliance, circumferential wall tension, stress, strain and elastic modulus were calculated. Atropine resulted in an increase in the oesophageal wall compliance during isobaric distension, but no change in compliance was observed during isovolumic distension. The stress-strain relationship was found to be linear during both types of distension, before as well as after atropine. The Young's modulus, which is the slope of a linear stress-strain relationship, was significantly higher after atropine in the isovolumic study but not in the isobaric study. The stress-strain relationship of the active component (muscle contraction) was different during isovolumic and isobaric distensions but the passive components were similar. The passive and active stress-strain relationships of the human oesophagus resemble those of other soft biological tissues. Furthermore, the method of oesophageal distension has significant influence on the active but not the passive biomechanical properties due to a strain-rate effect. PMID- 12562938 TI - A novel chloride conductance activated by extracellular ATP in mouse parotid acinar cells. AB - Salivary gland fluid secretion is driven by transepithelial Cl- movement involving an apical Cl- channel whose molecular identity remains unknown. Extracellular ATP (ATP(o)) has been shown to activate a Cl- conductance (I(ATPCl)) in secretory epithelia; to gain further insight into I(ATPCl) in mouse parotid acinar cells, we investigated the effects of ATP(o) using the whole-cell patch-clamp technique. ATP(o) and 2'- and 3'-O-(4-benzoylbenzoyl)adenosine 5' triphosphate triethylammonium salt (Bz-ATP) produced concentration-dependent, time-independent Cl- currents with an EC50 of 160 and 15 microM, respectively. I(ATPCl) displayed a selectivity sequence of SCN- > I- = NO3- > Cl- > glutamate, similar to the Cl- channels activated by Ca2+, cAMP and cell swelling in acinar cells. In contrast, I(ATPCl) was insensitive to pharmacological agents that are known to inhibit these latter Cl- channels, was independent of Ca2+ and was not regulated by cell volume. Moreover, the I(ATPCl) magnitude from wild-type animals was comparable to that from mice with null mutations in the Cftr, Clcn3 and Clcn2 Cl- channel genes. Taken together, our results demonstrate that I(ATPCl) is distinct from the channels described previously in acinar cells. The activation of I(ATPCl) by Bz-ATP suggests that P2 nucleotide receptors are involved. However, inhibition of G-protein activation with GDP-beta-S failed to block I(ATPCl), and Cibacron Blue 3GA and 4,4'-diisothyocyanostilbene-2,2'-disulphonic disodium salt selectively inhibited the Na+ currents (presumably through P2X receptors) without altering I(ATPCl), suggesting that neither P2Y nor P2X receptors are likely to be involved in I(ATPCl) activation. We conclude that I(ATPCl) is not associated with Cl- channels previously characterized in mouse parotid acinar cells, nor is it dependent on P2 nucleotide receptor stimulation. I(ATPCl) expressed in acinar cells reflects the activation of a novel ATP-gated Cl- channel that may play an important physiological role in salivary gland fluid secretion. PMID- 12562939 TI - Melatonin modulates the light-induced sympathoexcitation and vagal suppression with participation of the suprachiasmatic nucleus in mice. AB - In mammals, the autonomic nervous system mediates the central circadian clock oscillation from the suprachiasmatic nucleus (SCN) to the peripheral organs, and controls cardiovascular, respiratory and gastrointestinal functions. The present study was conducted in mice to address whether light signals conveyed to the SCN can control peripheral autonomic functions, and further examined the impact of centrally administered melatonin on peripheral autonomic functions via activation of melatonin receptor signalling. In vivo electrophysiological techniques were performed in anaesthetised, open-chest and artificially ventilated mice whilst monitoring the arterial blood pressure and heart rate. Light induced an increase of the renal sympathetic nerve activity, arterial blood pressure and heart rate immediately after lights on. Conversely, light rapidly suppressed the gastric vagal parasympathetic nerve activity, which was affected neither by hepatic vagotomy nor by total subdiaphragmatic vagotomy. These autonomic responses were mediated by the SCN since bilateral SCN lesion totally abolished the light-evoked neuronal and cardiovascular responses. Melatonin administered intracerebroventricularly (I.C.V.) attenuated the sympathetic and vagal nerve activities in a dose-dependent manner with a threshold of 0.1 ng and these effects were blocked by I.C.V. pre-treatment of the competitive melatonin receptor antagonist luzindole. These results suggest that light induces sympathoexcitation and vagal suppression through the SCN and that melatonin modulates the light-induced autonomic responses via activation of the central melatonin receptor signalling. PMID- 12562940 TI - Postnatal cardiovascular function after manipulation of fetal growth by embryo transfer in the horse. AB - This study used between-breed embryo transfer in the horse to investigate the effects of maternal size and uterine capacity on fetal growth and postnatal cardiovascular and neuroendocrine functions. Equine embryos were transferred to establish eight Thoroughbred-in-Thoroughbred (TinT), seven Pony-in-Pony (PinP), five Thoroughbred-in-Pony (TinP) and eight Pony-in-Thoroughbred (PinT), pregnancies. Maternal and foal weights and placental microscopic area were measured at birth. At 6 days of postnatal life, arterial blood pressure and heart rate were monitored and blood samples were taken for hormone analysis before, during and after a 10 min period of nitroprusside-induced hypotension. Values for maternal and foal weights and placental area at birth were larger in TinT than in PinP pregnancies (P < 0.05). PinT pregnancies resulted in larger placentae and heavier foals relative to PinP (P < 0.05). TinP had smaller placentae and lighter foals relative to TinT (P < 0.05). Growth-enhanced (PinT) foals showed elevated basal arterial blood pressure and baroreflex threshold, reduced baroreflex sensitivity, diminished plasma catecholamine responses to acute stress, and increased cortisol responsiveness to ACTH. Conversely, growth-restricted (TinP) foals showed no change in basal arterial blood pressure, baroreflex threshold or adrenocortical responsiveness to ACTH, but had enhanced baroreflex sensitivity and augmented plasma catecholamine responses to acute stress. The data show that fetal growth acceleration as well as fetal growth restriction, resulting from between-breed embryo transfer in the horse, leads to altered postnatal regulation of blood pressure and the circulating concentrations of cortisol. These findings suggest that deviations in the pattern and rate of fetal growth both above and below the normal trajectory may influence cardiovascular function in postnatal life. PMID- 12562941 TI - Adaptation of ovine fetal pancreatic insulin secretion to chronic hypoglycaemia and euglycaemic correction. AB - Fetal pancreatic adaptations to relative hypoglycaemia, a characteristic of intra uterine growth restriction, may limit pancreatic beta-cell capacity to produce and/or secrete insulin. The objective of this study was to measure beta-cell responsiveness in hypoglycaemic (H) fetal sheep and ascertain whether a 5 day euglycaemic recovery period would restore insulin secretion capacity. Glucose stimulated insulin secretion (GSIS) was measured in euglycaemic (E) control fetuses, fetuses made hypoglycaemic for 14 days, and in a subset of 14-day hypoglycaemic fetuses returned to euglycaemia for 5 days (R fetuses). Hypoglycaemia significantly decreased plasma insulin concentrations in H (0.13 +/ 0.01 ng ml(-1)) and R fetuses (0.11 +/- 0.01 ng ml(-1)); insulin concentrations returned to euglycaemic control values (0.30 +/- 0.01 ng ml(-1)) in R fetuses (0.29 +/- 0.04 ng ml(-1)) during their euglycaemic recovery period. Mean steady state plasma insulin concentration during the GSIS study was reduced in H fetuses (0.40 +/- 0.07 vs. 0.92 +/- 0.10 ng ml(-1) in E), but increased (P < 0.05) in R fetuses (0.73 +/- 0.10 ng ml(-1)) to concentrations not different from those in the E group. Nonlinear modelling of GSIS showed that response time was greater (P < 0.01) in both H (15.6 +/- 2.8 min) and R (15.4 +/- 1.5 min) than in E fetuses (6.3 +/- 1.1 min). In addition, insulin secretion responsiveness to arginine was reduced by hypoglycaemia (0.98 +/- 0.11 ng ml(-1) in H vs. 1.82 +/- 0.17 ng ml( 1) in E, P < 0.05) and did not recover (1.21 +/- 0.15 ng ml(-1) in R, P < 0.05 vs. E). Thus, a 5 day euglycaemic recovery period from chronic hypoglycaemia reestablished GSIS to normal levels, but there was a persistent reduction of beta cell responsiveness to glucose and arginine. We conclude that programming of pancreatic insulin secretion responsiveness can occur in response to fetal glucose deprivation, indicating a possible mechanism for establishing, in fetal life, a predisposition to type 2 diabetes. PMID- 12562942 TI - Maternal protein restriction in the rat impairs resistance artery but not conduit artery function in pregnant offspring. AB - Dietary protein restriction during gestation has been shown to produce vascular dysfunction in pregnant rats and hypertension in their offspring. However, no studies have to date examined the effects of such 'programming' on the vascular function of female offspring when they in turn become pregnant. We have therefore studied isolated conduit and resistance artery function from pregnant female offspring of control (C, 18 % casein) and protein-restricted (PR, 9 % casein) pregnant dams. There were no differences in birth weight, weight gain during pregnancy, litter size, fetal weight, placental weight, fetal : placental weight ratio or organ weights between the C and PR groups. In isolated mesenteric arteries, the vasodilatation in response to the endothelial-dependent vasodilator acetylcholine (ACh) and the beta-adrenoceptor agonist isoprenaline was decreased in the PR group, while there were no differences in the constriction in response to potassium (125 mM) or the alpha1-adrenoceptor agonist phenylephrine (PE). No differences in any responses were seen in the isolated thoracic aorta. We conclude that dietary protein restriction in pregnancy programmes vasodilator dysfunction in isolated resistance arteries of female offspring when they become pregnant, but does not affect conduit arteries. PMID- 12562943 TI - Glucocorticoid exposure at the dose used clinically alters cytoskeletal proteins and presynaptic terminals in the fetal baboon brain. AB - Glucocorticoids have been used for 30 years to accelerate fetal lung maturation in human pregnancy at risk of preterm delivery. Exposure to inappropriate levels of steroid, however, leads to altered maturation of the cardiovascular, metabolic and central nervous systems. The effects of betamethasone on neuronal development and function were determined in the fetal baboon brain by examination of cytoskeletal microtubule associated proteins (MAPs) and the presynaptic marker protein synaptophysin. At 0.73 gestation, commencing 28 weeks of gestation, pregnant baboons received four doses of saline (n = 8) or 87.5 microg (kg body weight)(-1) betamethasone I.M. (n = 7) 12 h apart. This dose is equivalent to 12 mg betamethasone administered daily over two consecutive days to a 70 kg woman. Baboons underwent Caesarean section 12 h after the last injection. Paraffin sections of the fetal neocortex and the underlying white matter were labelled immunohistochemically against MAP1B, MAP2abc, MAP2ab and synaptophysin and stained histochemically with hematoxylin-eosin and silver. Tissue staining was quantified morphometrically. Betamethasone exposure resulted in decreased immunoreactivity (IR) of MAP1B by 34.3 % and MAP2abc by 34.1 % (P < 0.05). Loss of MAP2 IR was due to loss of IR of the juvenile isoform MAP2c (P < 0.05). MAP1B and MAP2c are involved in neuritogenesis and neuronal plasticity. Synaptophysin IR was reduced by 51.8 % (P < 0.01). These changes might reflect functional neuronal disturbances because they were not accompanied by an alteration of the density of neurofibrils or neuronal necrosis. These results are in agreement with earlier findings of alterations of cytoskeletal proteins and presynaptic terminals in the fetal sheep brain after betamethasone infusion directly to the fetus and support a common effect of inappropriate fetal exposure to glucocorticoids on neuronal cytoskeleton and synapses in mammalian species. PMID- 12562944 TI - External perturbation of the trunk in standing humans differentially activates components of the medial back muscles. AB - During voluntary arm movements, the medial back muscles are differentially active. It is not known whether differential activity also occurs when the trunk is perturbed unpredictably, when the earliest responses are initiated by short latency spinal mechanisms rather than voluntary commands. To assess this, in unpredictable and self-initiated conditions, a weight was dropped into a bucket that was held by the standing subject (n = 7). EMG activity was recorded from the deep (Deep MF), superficial (Sup MF) and lateral (Lat MF) lumbar multifidus, the thoracic erector spinae (ES) and the biceps brachii. With unpredictable perturbations, EMG activity was first noted in the biceps brachii, then the thoracic ES, followed synchronously in the components of the multifidus. During self-initiated perturbations, background EMG in the Deep MF increased two- to threefold, and the latency of the loading response decreased in six out of the seven subjects. In Sup MF and Lat MF, this increase in background EMG was not observed, and the latency of the loading response was increased. Short-latency reflex mechanisms do not cause differential action of the medial back muscles when the trunk is loaded. However, during voluntary tasks the central nervous system exerts a 'tuned response', which involves discrete activity in the deep and superficial components of the medial lumbar muscles in a way that varies according to the biomechanical action of the muscle component. PMID- 12562945 TI - Alterations in slow-twitch muscle phenotype in transgenic mice overexpressing the Ca2+ buffering protein parvalbumin. AB - The purpose of this study was to determine whether induced expression of the Ca2+ buffering protein parvalbumin (PV) in slow-twitch fibres would lead to alterations in physiological, biochemical and molecular properties reflective of a fast fibre phenotype. Transgenic (TG) mice were generated that overexpressed PV in slow (type I) muscle fibres. In soleus muscle (SOL; 58 % type I fibres) total PV expression was 2- to 6-fold higher in TG compared to wild-type (WT) mice. Maximum twitch and tetanic tensions were similar in WT and TG but force at subtetanic frequencies (30 and 50 Hz) was reduced in TG SOL. Twitch time-to-peak tension and half-relaxation time were significantly decreased in TG SOL (time-to peak tension: 39.3 +/- 2.6 vs. 55.1 +/- 4.7 ms; half-relaxation time: 42.1 +/- 3.5 vs. 68.1 +/- 9.6 ms, P < 0.05 for TG vs. WT, respectively; n = 8-10). There was a significant increase in expression of type IIa myosin heavy chain (MHC) and ryanodine receptor at the mRNA level in TG SOL but there were no differences in MHC expression at the protein level and thus no difference in fibre type. Whole muscle succinate dehydrogenase activity was reduced by 12 +/- 0.4 % in TG SOL and single fibre glycerol-3-phosphate dehydrogenase activity was decreased in a subset of type IIa fibres. These differences were associated with a 64 % reduction in calcineurin activity in TG SOL. These data show that overexpression of PV, resulting in decreased calcineurin activity, can alter the functional and metabolic profile of muscle and influence the expression of key marker genes in a predominantly slow-twitch muscle with minimal effects on the expression of muscle contractile proteins. PMID- 12562946 TI - The dangerous road of catch-up growth. AB - Many epidemiological studies have now shown a strongly increased risk of developing type 2 diabetes and the metabolic syndrome in adults who as neonates showed signs of poor early (fetal and early postnatal) growth. The thrifty phenotype hypothesis was proposed to provide a conceptual and experimentally testable basis of these relationships. We have used protein restriction of rat dams, as a means to test this hypothesis. In vivo and in vitro studies of the growth-restricted offspring of such pregnancies have provided findings showing remarkable parallels with the human conditions. Permanent changes in the expression of regulatory proteins in liver, muscle and adipose tissue provide at least part of the explanation of the changes observed and offer potential markers for testing in the human context. These studies have also raised the question as to whether 'catch up' growth following early growth retardation may add to the risks posed by this early handicap. Male rats growth-retarded during fetal life and cross-fostered shortly after birth to normal lactating dams reach normal body and organ weights by weaning but have a reduced longevity. This finding raises the possibility that catch up growth, whilst potentially beneficial in the short term, may be detrimental to long-term survival. Human epidemiological studies may point in the same direction. Work by others on other models of early growth restriction have produced similar, although more limited, data. These findings raise the interesting possibility that the response to fetal stress, be it nutritional or other, may evoke a somewhat restricted and uniform pattern of adaptive response. PMID- 12562947 TI - The effect of birth weight on hypothalamo-pituitary-adrenal axis function in juvenile and adult pigs. AB - Programming of the hypothalamo-pituitary-adrenal (HPA) axis during prenatal and early postnatal life may explain, in part, the association between low birth weight (BW) and the increased incidence of cardiovascular and metabolic disease in later life. This study examined the effect of natural variations in BW on HPA axis function in juvenile and adult pigs. Low (< 1.47 kg) and high (> 1.53 kg) BW pure-bred Large White piglets from 15 litters were studied at 3 (n = 47) and 12 (n = 17) months of age. At each age, HPA axis function was tested by hypoglycaemic challenge (I.V. insulin; 0.5 IU (kg body weight)(-1)) and ACTH challenge (I.V. Synacthen, 2 microg (kg body weight)(-1)). At 3 months of age, adrenal size, the ratio of adrenal cortical to medullary area and stimulated cortisol concentrations were elevated in pigs that were of low BW and that remained small after birth. At 12 months of age, thinness at birth was associated with elevated adrenal responsiveness to insulin-induced hypoglycaemia. These results are consistent with the hypothesis that impaired fetal and early postnatal growth are associated with altered HPA axis function in later life. PMID- 12562948 TI - Placental glucose transport in growth-restricted pregnancies induced by overnourishing adolescent sheep. AB - Glucose clamp procedures were used to determine whether the slowing of fetal growth during the final third of gestation in overnourished adolescent ewes is due to a reduction in placental glucose transport capacity. Singleton pregnancies to a single sire were established by embryo transfer and thereafter adolescent dams were offered a high (n = 11) or moderate (n = 7) nutrient intake. Studies were conducted at 130 +/- 0.5 days gestation. Uterine and umbilical blood flows were studied by the steady-state transplacental diffusion technique and glucose fluxes quantified by the Fick principle. To determine the relationship between the transplacental glucose gradient and umbilical (fetal) glucose uptake, studies were conducted with maternal arterial glucose clamped at 5 micromol ml(-1) and fetal glucose at spontaneously occurring and two additional higher levels. Maternal body weight gain during gestation averaged 282 and 57 g day(-1) for high and moderate-intake dams, respectively. Total placentome weight (209 +/- 23 vs. 386 +/- 34 g) and fetal weight (3072 +/- 266 vs. 4670 +/- 196 g) were lower (P < 0.001) in high- than in moderate-intake groups. The growth-restricted pregnancies in the high-intake dams were associated with reduced uterine (P < 0.05) and umbilical (P < 0.02) blood flows and, in the non-perturbed state, the fetuses were relatively hypoxic (2.1 vs. 3.0 micromol ml(-1), P < 0.05) and hypoglycaemic (0.90 vs. 1.31 micromol ml(-1), P < 0.002). Linear regression analysis of umbilical glucose uptake at three steady-state uterine-umbilical arterial transplacental plasma glucose concentration gradients revealed that absolute placental glucose transport capacity was lower in high- than in moderate-intake dams (mean slope, 0.8 vs. 1.5 dl min(-1), P < 0.05; and mean intercept, 1.84 vs. 3.40 micromol ml(-1)). However, glucose transfer capacity was not different between the two groups when expressed on a placental weight-specific basis. This confirms that the small size of the placenta per se is the major limitation to placental glucose transfer in the overnourished adolescent pregnant sheep. PMID- 12562949 TI - Augmentation of coronary conductance in adult sheep made anaemic during fetal life. AB - Maximal coronary conductance with adenosine in anaemic fetal sheep is twice that of non-anaemic fetuses. To investigate whether this increase in conductance persists into adulthood we studied twin sheep as fetuses and again as adults. Nine anaemic fetuses (118 days gestation) underwent isovolaemic haemorrhage for 18.0 +/- 4.6 days (means +/- S.D.) during which time the haematocrit was reduced from 39.9 +/- 5.2 % to 16.3 +/- 3.4 % and oxygen content from 8.6 +/- 1.3 to 2.3 +/- 0.2 ml dl-1. At 138 days the anaemic fetuses were transfused; at delivery the haematocrit was 29.3 +/- 6.8 % compared to nine control fetuses in which the haematocrit was 38.5 +/- 4.3 %. The weight at delivery was 3.5 +/- 0.36 kg in the anaemic fetuses vs. 4.2 +/- 0.83 kg in controls. Twenty-eight weeks later, we placed an occluder on the descending thoracic aorta and inferior vena cava, a flow probe around the proximal left circumflex coronary artery, and catheters in the left atrial appendage, jugular and carotid vessels. Maximal coronary conductance was determined in the adults by recording coronary blood flow as driving pressure was altered by inflating the occluders while adenosine was infused into the left atrium. Right atrial, left atrial, systolic and mean arterial pressures, systemic vascular resistance and haematocrit were not different between 'in utero anaemic' and control adults. The adults that were anaemic in utero weighed less than the controls 39.4 +/- 4.6 kg vs. 45.0 +/- 5.6 kg. Maximal conductance was greater in the adults that were anaemic in utero: 11.2 +/- 4.0 ml min(-1) (100 g)(-1) mmHg-1 as compared to 6.1 +/- 1.8 ml min(-1) (100 g)(-1) mmHg(-1) in the controls. Vascular reactivity of the mesenteric arteries was not different. These data suggest that coronary conductance can be modified in utero by anaemia (high flow and hypoxaemia) and that the remodelled coronary tree persists to adulthood. PMID- 12562950 TI - Surgery in the human fetus: the future. AB - Fetal surgery was born of clinical necessity. Observations by pediatric surgeons and neonatologists of neonates that were born with irreversible organ damage led to the conclusion that one possible approach to prevent this alteration of developmental physiology, was fetal surgical intervention. This led to experimental validation of the pathophysiology of specific fetal defects in animal models and to the development of techniques for their prenatal surgical correction. The demonstration in animal models that the correction of an anatomical defect could reverse the associated pathophysiology led to the first systematic application of fetal surgery at the University of California, San Francisco, in the early 1980s. Since that time, fetal surgery has been applied in only a few centres and has remained relatively limited in scope. Nevertheless, there has been a dramatic improvement in our ability to diagnose, select and safely operate on an expanding number of fetal anomalies. The purpose of this article is to briefly summarize the present status of fetal surgery and to speculate about what may be in store for the future. Inherent in such an effort is a definition of what constitutes fetal surgery. In this discussion I will take considerable latitude with the definition of what constitutes fetal surgery in the future, as it is my belief that technological progress in a number of areas will result in dramatic changes in the practice and perception of fetal surgery. PMID- 12562951 TI - Chronic hypoxia causes angiogenesis in addition to remodelling in the adult rat pulmonary circulation. AB - Chronic hypoxia caused by migration of native sea-level dwellers to high altitude or chronic lung disease leads to the development of increased pulmonary vascular resistance and pulmonary hypertension. This altitude-induced hypertension offers no obvious benefit and may indeed be maladaptive. A major mechanism thought to contribute to the development of pulmonary hypertension is hypoxia-induced loss of small blood vessels, sometimes termed rarefaction or pruning. More recent evidence caused us to question this widely accepted concept including the potent angiogenic effect of chronic hypoxia in all other vascular beds and the demonstration that new vessels can form in the pulmonary circulation when stimulated by chronic infection and lung resection. We tested the hypothesis that chronic environmental hypoxia causes angiogenesis in the adult pulmonary circulation by using stereological techniques combined with confocal microscopy to examine the resultant changes in pulmonary vascular structure in rats. We found that chronic hypoxia resulted in increased total pulmonary vessel length, volume, endothelial surface area and number of endothelial cells in vivo. This is the first reported demonstration of hypoxia-induced angiogenesis in the mature pulmonary circulation, a structural adaptation that may have important beneficial consequences for gas exchange. These findings imply that we must revise the widely accepted paradigm that hypoxia-induced loss of small vessels is a key structural change contributing to the development of pulmonary hypertension in high altitude adaptation and chronic lung disease. PMID- 12562952 TI - Allosteric regulation and spatial distribution of kainate receptors bound to ancillary proteins. AB - A diverse range of accessory proteins regulates the behaviour of most ligand- and voltage-gated ion channels. For glutamate receptor 6 (GluR6) kainate receptors, two unrelated proteins, concanavalin-A (Con-A) and postsynaptic density protein 95 (PSD-95), bind to extra- and intracellular domains, respectively, but are reported to exert similar effects on GluR6 desensitization behaviour. We have tested the hypothesis that distinct allosteric binding sites control GluR6 receptors via a common transduction pathway. Rapid agonist application to excised patches revealed that neither Con-A nor PSD-95 affect the onset of desensitization. The rate of desensitization elicited by 10 mM L-glutamate was similar in control (taufast = 5.5 +/- 0.4 ms), Con-A-treated patches (taufast = 6.1 +/- 0.5 ms) and patches containing PSD-95 and GluR6 receptors (taufast = 4.7 +/- 0.6 ms). Likewise, the time course of recovery from GluR6 desensitization was similar in both control and Con-A conditions, whereas PSD-95 accelerated recovery almost twofold. Peak and steady-state (SS) dose-response relationships to glutamate were unchanged by lectin treatment (e.g. control, EC50(SS) = 31 +/- 28 microM vs Con-A, EC50(SS) = 45 +/- 9 microM, n = 6), suggesting that Con-A does not convert non-conducting channels with high agonist affinity into an open conformation. Instead, we demonstrate that the effects of Con-A on macroscopic responses reflect a shift in the relative contribution of different open states of the channel. In contrast, the effect of PSD-95 on recovery behaviour suggests that the association between kainate receptors and cytoskeletal proteins regulates signalling at glutamatergic synapses. Our results show that Con-A and PSD-95 regulate kainate receptors via distinct allosteric mechanisms targeting selective molecular steps in the transduction pathway. PMID- 12562953 TI - Acute shifts of baroreflex control of renal sympathetic nerve activity induced by treadmill exercise in rats. AB - The present study aimed to investigate whether there was a resetting of the baroreflex control of renal sympathetic nerve activity (RSNA) and heart rate (HR) during exercise. Wistar female rats (n = 11) were chronically implanted with catheters for the measurement of systemic arterial (Pa) and central venous pressures and with electrodes for measurement of RSNA and electrocardiogram (ECG) at least 3 days before study. The baroreflex curve for RSNA was determined by changing Pa using rapid intravenous infusions of phenylephrine and nitroprusside. The baroreflex response curves for RSNA and HR were characterized by an inverse sigmoid function curve from which the response range, gain, centering point and minimum response were estimated. Exercise shifted the Pa-RSNA baroreflex curve upward and to the right and was associated with increases in response range of 122 +/- 44 % (P < 0.05), maximum response of 173 +/- 40 % (P < 0.05), maximum gain of 149 +/- 66 % (P < 0.05) and midpoint pressure of 15 +/- 5 mmHg (P < 0.05) compared with the pre-exercise level. After cessation of exercise, the Pa-RSNA baroreflex curve was suppressed vertically with a significant decrease in maximum response of 57 +/- 14 % (P < 0.05) compared with the pre-exercise level. These data suggest that the right-upward shift of baroreflex control of sympathetic nerve activity may play a critical role in raising and stabilizing Pa during exercise. The suppression of the baroreflex control of sympathetic nerve activity may partly explain the post-exercise inhibition of sympathetic nerve activity and contribute to the post-exercise hypotension. PMID- 12562954 TI - Activity of brainstem respiratory neurones just before the expiration-inspiration transition in the rat. AB - Inspiratory activity of the hypoglossal nerve (XIIn) often precedes that of the phrenic nerve (PHRn). By manipulating artificial respiration, this preceding activity (pre-I XIIn activity) can be lengthened or isolated prematurely (decoupled XIIn activity) without developing into overt PHRn-associated inspiratory bursts. We hypothesized that these pre-I and decoupled XIIn activities, collectively termed 'XIIn-w/o-PHRn activity', reflect certain internal states of the respiratory centre at the period just prior to the transition from the expiratory phase to the inspiratory phase. In decerebrate, neuromuscularly blocked and artificially ventilated rats, the firing properties of medullary respiratory neurones were examined during the period of the XIIn-w/o PHRn activity. The majority of the inspiratory neurones examined could be classified into two types: one was active (XIIn-type) and the other was inactive (PHRn-type) during the XIIn-w/o-PHRn period. On the other hand, augmenting expiratory (E-AUG) neurones of the Botzinger complex (BOT) and the caudal ventral respiratory group (VRG) fired intensively during this period. Their firing stopped at the onset of the overt inspiratory bursts in the XIIn and PHRn, suggesting that BOT E-AUG neurones inhibit PHRn-type, but not XIIn-type, inspiratory neurones. We hypothesize that XIIn-type inspiratory activity facilitates the phase change from expiration to inspiration, through activation of certain inspiratory neurones that inhibit the firing of BOT E-AUG neurones and generation of the overt inspiratory bursts in XIIn-type and PHRn-type inspiratory neurones. PMID- 12562955 TI - Local routes revisited: the space and time dependence of the Ca2+ signal for phasic transmitter release at the rat calyx of Held. AB - During the last decade, advances in experimental techniques and quantitative modelling have resulted in the development of the calyx of Held as one of the best preparations in which to study synaptic transmission. Here we review some of these advances, including simultaneous recording of pre- and postsynaptic currents, measuring the Ca2+ sensitivity of transmitter release, reconstructing the 3-D anatomy at the electron microscope (EM) level, and modelling the buffered diffusion of Ca2+ in the nerve terminal. An important outcome of these studies is an improved understanding of the Ca2+ signal that controls phasic transmitter release. This article illustrates the spatial and temporal aspects of the three main steps in the presynaptic signalling cascade: Ca2+ influx through voltage gated calcium channels, buffered Ca2+ diffusion from the channels to releasable vesicles, and activation of the Ca2+ sensor for release. Particular emphasis is placed on how presynaptic Ca2+ buffers affect the Ca2+ signal and thus the amplitude and time course of the release probability. Since many aspects of the signalling cascade were first conceived with reference to the squid giant presynaptic terminal, we include comparisons with the squid model and revisit some of its implications. Whilst the characteristics of buffered Ca2+ diffusion presented here are based on the calyx of Held, we demonstrate the circumstances under which they may be valid for other nerve terminals at mammalian CNS synapses. PMID- 12562956 TI - Enhanced nitric oxide activity offsets peripheral vasoconstriction during acute hypoxaemia via chemoreflex and adrenomedullary actions in the sheep fetus. AB - We tested the hypothesis that enhanced nitric oxide (NO) opposes fetal peripheral vasoconstrictor responses to acute hypoxaemia via actions involving the carotid chemoreflex and the adrenal medulla. The hypothesis was tested in the late gestation ovine fetus using a novel NO clamp technique, which involves fetal combined treatment with the NO synthase inhibitor, L-NAME, and the NO donor, sodium nitroprusside. In contrast to treatment with L-NAME alone, combined fetal treatment with L-NAME and nitroprusside prevents generalized vasoconstriction and pronounced hypertension, not only maintaining basal cardiovascular function, but also permitting blockade of the de novo synthesis of NO during hypoxaemia while compensating for the tonic production of the gas. Under general anaesthesia, seven sheep fetuses were surgically prepared with catheters and a femoral Transonic flow probe. Five days after surgery, fetuses were subjected to a 3 h protocol: 1 h normoxia, 1 h hypoxaemia and 1 h recovery. Fetal hypoxaemia was induced during either fetal infusion with saline or treatment with the NO clamp. During saline infusion, fetuses responded to hypoxaemia with transient bradycardia, femoral vasoconstriction and increases in plasma noradrenaline and adrenaline. During fetal treatment with the NO clamp, bradycardia persisted and there were greater peripheral vasoconstrictor and catecholaminergic responses to hypoxaemia. Further analysis showed that NO clamp treatment enhanced the chemoreflex component of the fetal cardiovascular defence to acute hypoxaemia. These data support the hypothesis that enhanced NO synthesis during acute hypoxaemia offsets fetal peripheral vasoconstrictor responses to hypoxaemia via chemoreflex and adrenomedullary actions. PMID- 12562957 TI - Prenatal dexamethasone leads to both endothelial dysfunction and vasodilatory compensation in sheep. AB - We investigated long-term cardiovascular effects in the offspring of sheep exposed to prenatal dexamethasone (DM). We assessed in vitro vascular responsiveness and evaluated endothelial nitric oxide synthase (eNOS) message and protein levels in femoral muscle removed from 5-month-old sheep. Dexamethasone was administered i.m. to pregnant ewes as 3 weekly courses (4 x 2 mg at 12 h intervals), starting on day 103 of gestation (term approximately 149 days). Ewes were allowed to lamb. At 5 months of age a carotid catheter was placed for blood pressure measurement and hamstring muscle was removed from the lambs under general anaesthesia. We demonstrate that following prenatal DM exposure in the 5 month-old offspring: (1) blood pressure is unchanged; (2) as previously reported in the fetus, sensitivity to endothelin-1 (ET) is increased; (3) acetylcholine induced relaxation is increased; (4) L-NAME suppressible vasodilatory response to ET is abolished; (5) there is no change in endothelium-independent vasodilatation; and (6) there is no change in eNOS RNA and protein levels, when compared to saline treated controls. We speculate that decreased agonist-induced NO release is not due to alteration in gene expression, but is more likely to be a post-transcriptional event. In summary, the lack of a difference in resting mean arterial pressure (MAP) between DM and control lambs indicates that the compensation we have previously demonstrated in the fetus following glucocorticoid exposure persists to 5 months postnatal age. Compensation is likely due to non-NO-dependent mechanisms, since no evidence was found of upregulated NOS. PMID- 12562958 TI - Basal leg blood flow in healthy women is related to age and hormone replacement therapy status. AB - Basal leg blood flow declines with age in healthy men, an effect that is mediated by augmented sympathetic vasoconstriction. However, in women the presence or absence of oestrogen and selective use of hormone replacement therapy (HRT) may alter these relationships with ageing. We studied 103 healthy women: 73 postmenopausal (41 HRT, mean +/- S.E.M. 61 +/- 1 years; 32 no-HRT, 63 +/- 2 years) and 30 premenopausal (29 +/- 1 years). Compared with the premenopausal controls, absolute femoral artery blood flow (duplex ultrasound) was 23 % lower (P < 0.001) in the postmenopausal no-HRT group, but only 13 % lower in the HRT group (P < 0.01). The age and HRT group differences in leg blood flow were consistently associated with differences in leg vascular conductance, but not with femoral artery lumen diameter, leg muscle sympathetic nerve activity or cardiac output (systemic arterial blood flow). Leg fat-free mass was smaller in the postmenopausal groups (P < 0.05). Femoral blood flow normalized for leg fat free mass was 17 % lower (P < 0.01) in the postmenopausal no-HRT compared with the premenopausal women, but was not different in the postmenopausal HRT and premenopausal groups. Femoral artery shear stress was similar in the postmenopausal HRT and premenopausal women, but was lower in the postmenopausal no-HRT group (P < 0.01). Basal whole-leg blood flow declines with age in healthy, oestrogen-deficient women, a phenomenon that is mediated primarily by reductions in leg vascular conductance. Among postmenopausal women, chronic HRT use is associated with augmented basal leg blood flow and vascular conductance. Leg blood flow normalized for leg fat-free mass is preserved with age in women taking chronic HRT. In contrast to men, differences in leg sympathetic vasoconstrictor nerve activity do not explain group differences in leg blood flow and vascular conductance with ageing in women. PMID- 12562959 TI - A mechanism underlying the sexually dimorphic ACTH response to lipopolysaccharide in rats: sex steroid modulation of cytokine binding sites in the hypothalamus. AB - It is well established that the hypothalamic-pituitary-adrenal responses to immune stressors are sexually dimorphic in rodents (females > males), but the underlying mechanism is still unclear. To investigate the mechanism, in this study we examined whether the sex steroid environment affects the following variables in male and female rats: (1) plasma levels of ACTH, interleukin (IL) 1beta, IL-6 and tumour necrosis factor-alpha (TNF-alpha) after systemic lipopolysaccharide (LPS) administration; (2) static concentrations of corticotropin-releasing hormone (CRH) and arginine vasopressin (AVP) in the mediobasal hypothalamus (MBH) and those of ACTH in the anterior pituitary (AP); and (3) the binding characteristics of IL-1beta, IL-6 and TNF-alpha in the MBH and AP. LPS-induced ACTH release was significantly higher in female than in male rats, and this sexual difference was abolished by performing gonadectomy in both sexes. Administration of physiological doses of testosterone and oestradiol to gonadectomized males and females, respectively, restored the altered ACTH responses to normal. Changes in the sex steroid milieu did not affect plasma cytokine responses to LPS, tissue contents of CRH, AVP and ACTH, or the IL-6 binding characteristics in the MBH and AP. However, the number of IL-1beta and TNF-alpha binding sites, but not their binding affinities, in the MBH showed significant changes according to altered sex hormone milieu, in the same direction as the LPS-induced ACTH response. These results suggest that the hypothalamic sensitivity to peripheral IL-1beta and TNF-alpha may be an important mechanism underlying the sexually dimorphic ACTH response to LPS in rats. PMID- 12562960 TI - Expression of IGF-I splice variants in young and old human skeletal muscle after high resistance exercise. AB - The mRNA expression of two splice variants of the insulin-like growth factor-I (IGF-I) gene, IGF-IEa and mechano growth factor (MGF), were studied in human skeletal muscle. Subjects (eight young, aged 25-36 years, and seven elderly, aged 70-82 years) completed 10 sets of six repetitions of single legged knee extensor exercise at 80 % of their one repetition maximum. Muscle biopsy samples were obtained from the quadriceps muscle of both the control and exercised legs 2.5 h after completion of the exercise bout. Expression levels of the IGF-I mRNA transcripts were determined using real-time quantitative RT-PCR with specific primers. The resting levels of MGF were significantly (approximately 100-fold) lower than those of the IGF-IEa isoform. No difference was observed between the resting levels of the two isoforms between the two subject groups. High resistance exercise resulted in a significant increase in MGF mRNA in the young, but not in the elderly subjects. No changes in IGF-IEa mRNA levels were observed as a result of exercise in either group. The mRNA levels of the transcription factor MyoD were greater at rest in the older subjects (P < 0.05), but there was no significant effect of the exercise bout. Electrophoretic separation of myosin heavy chain (MHC) isoforms showed the older subjects to have a lower (P < 0.05) percentage of MHC-II isoforms than the young subjects. However, no association was observed between the composition of the muscle and changes in the IGF-I isoforms with exercise. The data from this study show an attenuated MGF response to high resistance exercise in the older subjects, indicative of age-related desensitivity to mechanical loading. The data in young subjects indicate that the MGF and IGF-IEa isoforms are differentially regulated in human skeletal muscle. PMID- 12562961 TI - Isometric force and endurance in skeletal muscle of mice devoid of all known thyroid hormone receptors. AB - The importance of thyroid hormone receptors for isometric force, endurance and content of specific muscle enzymes was studied in isolated slow-twitch soleus and fast-twitch extensor digitorum longus (EDL) muscles in mice deficient in all known subtypes of thyroid hormone receptors (i.e. TR alpha1, beta1, beta2 and beta3). The weights of soleus and EDL muscles were lower in TR-deficient (TRalpha1-/-beta-/-) mice than in wild-type controls. The force per cross sectional area was not significantly different between TRalpha1-/-beta-/- and wild-type muscles. Soleus muscles of TRalpha1-/-beta-/- mice showed increased contraction and relaxation times and the force-frequency relationship was shifted to the left. Soleus muscles of TRalpha1-/-beta-/- mice were more fatigue resistant than wild-type controls. Protein analysis of TRalpha1-/-beta-/- soleus muscles showed a marked increase in expression of the slow isoform of the sarcoplasmic reticulum Ca2+ pump (SERCa2), whilst expression of the fast type (SERCa1) was decreased. There was also a major decrease in the alpha2-subunit of the Na+-K+ pump in TRalpha1-/-beta-/- soleus muscles. EDL muscles from TRalpha1-/ beta-/- and wild-type mice showed no significant difference in contraction and relaxation times, fatigue resistance and protein expression. In conclusion, the present data show changes in contractile characteristics of skeletal muscles of TRalpha1-/-beta-/- mice similar to those seen in hypothyroidism. We have previously shown that muscles of mice deficient in TRalpha1 or TRbeta display modest changes in muscle function. Thus, in skeletal muscle there seems to be functional overlap between TRalpha1 and TRbeta, so that the lack of one of the receptors to some extent can be compensated for by the presence of the other. PMID- 12562962 TI - Mast cell tryptase and proteinase-activated receptor 2 induce hyperexcitability of guinea-pig submucosal neurons. AB - Mast cells that are in close proximity to autonomic and enteric nerves release several mediators that cause neuronal hyperexcitability. This study examined whether mast cell tryptase evokes acute and long-term hyperexcitability in submucosal neurons from the guinea-pig ileum by activating proteinase-activated receptor 2 (PAR2) on these neurons. We detected the expression of PAR2 in the submucosal plexus using RT-PCR. Most submucosal neurons displayed PAR2 immunoreactivity, including those colocalizing VIP. Brief (minutes) application of selective PAR2 agonists, including trypsin, the activating peptide SL-NH2 and mast cell tryptase, evoked depolarizations of the submucosal neurons, as measured with intracellular recording techniques. The membrane potential returned to resting values following washout of agonists, but most neurons were hyperexcitable for the duration of recordings (> 30 min-hours) and exhibited an increased input resistance and amplitude of fast EPSPs. Trypsin, in the presence of soybean trypsin inhibitor, and the reverse sequence of the activating peptide (LR-NH2) had no effect on neuronal membrane potential or long-term excitability. Degranulation of mast cells in the presence of antagonists of established excitatory mast cell mediators (histamine, 5-HT, prostaglandins) also caused depolarization, and following washout of antigen, long-term excitation was observed. Mast cell degranulation resulted in the release of proteases, which desensitized neurons to other agonists of PAR2. Our results suggest that proteases from degranulated mast cells cleave PAR2 on submucosal neurons to cause acute and long-term hyperexcitability. This signalling pathway between immune cells and neurons is a previously unrecognized mechanism that could contribute to chronic alterations in visceral function. PMID- 12562963 TI - Analysis of the differential modulation of sulphonylurea block of beta-cell and cardiac ATP-sensitive K+ (K(ATP)) channels by Mg-nucleotides. AB - Sulphonylureas stimulate insulin secretion by binding with high-affinity to the sulphonylurea receptor (SUR) subunit of the ATP-sensitive potassium (K(ATP)) channel and thereby closing the channel pore (formed by four Kir6.2 subunits). In the absence of added nucleotides, the maximal block is around 60-80 %, indicating that sulphonylureas act as partial antagonists. Intracellular MgADP modulated sulphonylurea block, enhancing inhibition of Kir6.2/SUR1 (beta-cell type) and decreasing that of Kir6.2/SUR2A (cardiac-type) channels. We examined the molecular basis of the different response of channels containing SUR1 and SUR2A, by recording currents from inside-out patches excised from Xenopus oocytes heterologously expressing wild-type or chimeric channels. We used the benzamido derivative meglitinide as this drug blocks Kir6.2/SUR1 and Kir6.2/SUR2A currents, reversibly and with similar potency. Our results indicate that transfer of the region containing transmembrane helices (TMs) 8-11 and the following 65 residues of SUR1 into SUR2A largely confers a SUR1-like response to MgADP and meglitinide, whereas the reverse chimera (SUR128) largely endows SUR1 with a SUR2A-type response. This effect was not specific for meglitinide, as tolbutamide was also unable to prevent MgADP activation of Kir6.2/SUR128 currents. The data favour the idea that meglitinide binding to SUR1 impairs either MgADP binding or the transduction pathway between the NBDs and Kir6.2, and that TMs 8-11 are involved in this modulatory response. The results provide a basis for understanding how beta-cell K(ATP) channels show enhanced sulphonylurea inhibition under physiological conditions, whereas cardiac K(ATP) channels exhibit reduced block in intact cells, especially during metabolic inhibition. PMID- 12562964 TI - Endothelial nitric oxide synthase, caveolae and the development of atherosclerosis. AB - Early hypercholesterolaemia-induced vascular disease is characterized by an attenuated capacity for endothelial production of the antiatherogenic molecule nitric oxide (NO), which is generated by endothelial NO synthase (eNOS). In recent studies we have determined the impact of lipoproteins on eNOS subcellular localization and action, thereby providing a causal link between cholesterol status and initial abnormalities in endothelial function. We have demonstrated that eNOS is normally targeted to cholesterol-enriched caveolae where it resides in a signalling module. Oxidized low density lipoprotein (LDL; oxLDL) causes displacement of eNOS from caveolae by binding to endothelial cell CD36 receptors and by depleting caveolae cholesterol content, resulting in the disruption of eNOS activation. The adverse effects of oxLDL are fully prevented by high density lipoprotein (HDL) via binding to scavenger receptor BI (SR-BI), which is colocalized with eNOS in endothelial caveolae. This occurs through the maintenance of caveolae cholesterol content by cholesterol ester uptake from HDL. As importantly, HDL binding to SR-BI causes robust stimulation of eNOS activity in endothelial cells, and this process is further demonstrable in isolated endothelial cell caveolae. HDL also enhances endothelium- and NO-dependent relaxation in aortae from wild-type mice, but not in aortae from homozygous null SR-BI knockout mice. Thus, lipoproteins have potent effects on eNOS function in caveolae via actions on both membrane cholesterol homeostasis and the level of activation of the enzyme. These processes may be critically involved in the earliest phases of atherogenesis, which recent studies suggest may occur during fetal life. PMID- 12562965 TI - Interactions between neutrophils and macrophages promote macrophage killing of rat muscle cells in vitro. AB - Current evidence indicates that the physiological functions of inflammatory cells are highly sensitive to their microenvironment, which is partially determined by the inflammatory cells and their potential targets. In the present investigation, interactions between neutrophils, macrophages and muscle cells that may influence muscle cell death are examined. Findings show that in the absence of macrophages, neutrophils kill muscle cells in vitro by superoxide-dependent mechanisms, and that low concentrations of nitric oxide (NO) protect against neutrophil-mediated killing. In the absence of neutrophils, macrophages kill muscle cells through a NO-dependent mechanism, and the presence of target muscle cells causes a three fold increase in NO production by macrophages, with no change in the concentration of inducible nitric oxide synthase. Muscle cells that are co cultured with both neutrophils and macrophages in proportions that are observed in injured muscle show cytotoxicity through a NO-dependent, superoxide independent mechanism. Furthermore, the concentration of myeloid cells that is necessary for muscle killing is greatly reduced in assays that use mixed myeloid cell populations, rather than uniform populations of neutrophils or macrophages. These findings collectively show that the magnitude and mechanism of muscle cell killing by myeloid cells are modified by interactions between muscle cells and neutrophils, between muscle cells and macrophages and between macrophages and neutrophils. PMID- 12562966 TI - Sodium transport across the chorioallantoic membrane of porcine placenta involves the epithelial sodium channel (ENaC). AB - The properties of chorioallantoic membrane derived from Large White Landrace sows at 45, 65 and 100 days gestation are examined. Under short circuit conditions positive charge flows from fetal to maternal sides of the tissue. Na+ is shown to be the sole charge carrier as the short circuit current is inhibited reversibly by fetal applications of amiloride and replacement of Na+ by choline in the Ringer solution, and irreversibly by both fetal and maternal applications of ouabain. The initial short circuit current is smaller at day 100 compared to days 45 and 65. The dose responses to amiloride indicate that the epithelial sodium channel (ENaC) is involved in the movement of Na+ and that it is accessible on the fetal side of the tissue only. Immunostaining shows that the ENaC-alpha subunit is present in both the allantoic membrane and the trophoblast. Uptake studies using microvillous (apical) membrane vesicles suggest it is either inactive or only weakly active at this site. The trophoblast at day 100 has a higher content of ENaC than at days 45 and 65. This is the first report of the presence of ENaC in placental tissues. The effects of ouabain indicate the presence of a Na+ pump that is more readily inhibited by applications of the drug on the maternal aspect of the tissue than on the fetal side. Differential mechanisms may be present that would allow net movement of Na+ in either direction across the chorioallantoic membrane according to the changing demands of the developing fetus. PMID- 12562967 TI - Decreased surface tension of upper airway mucosal lining liquid increases upper airway patency in anaesthetised rabbits. AB - The obstructive sleep apnoea syndrome (OSA) is a disorder characterised by repetitive closure and re-opening of the upper airway during sleep. Upper airway luminal patency is influenced by a number of factors including: intraluminal air pressure, upper airway dilator muscle activity, surrounding extraluminal tissue pressure, and also surface forces which can potentially act within the liquid layer lining the upper airway. The aim of the present study was to examine the role of upper airway mucosal lining liquid (UAL) surface tension (gamma) in the control of upper airway patency. Upper airway opening (PO) and closing pressures (PC) were measured in 25 adult male, supine, tracheostomised, mechanically ventilated, anaesthetised (sodium pentabarbitone), New Zealand White rabbits before (control) and after instillation of 0.5 ml of either 0.9 % saline (n = 9) or an exogenous surfactant (n = 16; Exosurf Neonatal) into the pharyngeal airway. The gamma of UAL (0.2 microl) was quantified using the 'pull-off' force technique in which gamma is measured as the force required to separate two curved silica discs bridged by the liquid sample. The gamma of UAL decreased after instillation of surfactant from 54.1 +/- 1.7 mN m-1 (control; mean +/- S.E.M.) to 49.2 +/- 2.1 mN m-1 (surfactant; P < 0.04). Compared with control, PO increased significantly (P < 0.04; paired t test, n = 9) from 6.2 +/- 0.9 to 9.6 +/- 1.2 cmH2O with saline, and decreased significantly (P < 0.05, n = 16) from 6.6 +/- 0.4 to 5.5 +/ 0.6 cmH2O with surfactant instillation. Findings tended to be similar for PC. Change in both PO and PC showed a strong positive correlation with the change in gamma of UAL (both r > 0.70, P < 0.001). In conclusion, the patency of the upper airway in rabbits is partially influenced by the gamma of UAL. These findings suggest a role for UAL surface properties in the pathophysiology of OSA. PMID- 12562968 TI - Modulation of AMPA receptors by cAMP-dependent protein kinase in preBotzinger complex inspiratory neurons regulates respiratory rhythm in the rat. AB - We hypothesize that phosphorylation of AMPA receptors or associated synaptic proteins modulates the excitability of respiratory neurons in the preBotzinger Complex (preBotC), affecting respiratory rhythm. Using neonatal rat medullary slices that spontaneously generate respiratory rhythm, we examined the role of the cAMP-PKA pathway (PKA: cAMP-dependent protein kinase) in modulating glutamatergic synaptic transmission, the excitability of inspiratory neurons in the preBotC and respiratory rhythm. Microinjection of forskolin, an activator of adenylate cyclase, into the preBotC with or without the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), decreased the period (increased the frequency) of respiratory-related rhythmic motor output in the hypoglossal nerve (XIIn) to 84 % (without IBMX) and to 72 % (with IBMX) of the pre-injection baseline. In the presence of MK-801, a non-competitive NMDA receptor antagonist, microinjection of forskolin plus IBMX decreased the period to 66 % of baseline levels. Microinjection of Rp-adenosine 3',5'-cyclic monophosphothioate (Rp cAMPS), a PKA inhibitor, increased the period to 145 % of baseline levels. Concurrent microinjection of Rp-cAMPS and forskolin had no effect on the period. Bath application of 7beta-deacetyl-7beta-[gamma-(morpholino)butyryl]-forskolin hydrochloride (7Db-forskolin, a water-soluble derivative of forskolin): (1) decreased the period to 67 % of baseline levels without affecting the amplitude of integrated XIIn inspiratory discharge, (2) induced a tonic inward current of 29 pA and enhanced inspiratory drive current (the amplitude increased to 183 % and the integral increased to 184 % of baseline) in voltage-clamped (holding potential = -60 mV) preBotC inspiratory neurons and (3) increased the frequency to 195 % and amplitude to 118 % of spontaneous excitatory postsynaptic currents (sEPSCs) during expiratory periods. Dideoxy-forskolin did not have these effects. Intracellular perfusion with the catalytic subunit of PKA (cPKA) into preBotC inspiratory neurons progressively enhanced inspiratory drive currents and, in the presence of TTX, increased the inward currents induced by local ejection of AMPA; the latter currents were blocked by 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo benzo[f]quinoxaline-7-sulphonamide (NBQX, an AMPA/kainate receptor antagonist). The effects of cPKA were blocked by co-application of PKA inhibitor (6-22) amide (PKI). These results suggest that phosphorylation of postsynaptic AMPA receptors through the cAMP-PKA pathway modulates both tonic and phasic excitatory amino acid synaptic transmission and excitability of inspiratory neurons in the preBotC and, therefore, regulates respiratory rhythm. Moreover, the basal level of endogenous PKA activity appears to be a determinant of resting respiratory frequency. PMID- 12562969 TI - Changes in respiratory control after three hours of isocapnic hypoxia in humans. AB - Despite the obvious role of hypoxia in eliciting respiratory acclimatisation in humans, the function of the peripheral chemoreflex is uncertain. We investigated this uncertainty using 3 h of isocapnic hypoxia as a stimulus (end-tidal PCO2, 0.5-1.0 mmHg above eucapnia; end-tidal PO2, 50 mmHg), hypothesising that this stimulus would induce an enhancement of the peripheral chemoreflex ventilatory response to hypoxia. Current evidence conflicts as to whether this enhancement is mediated by an increase in the sensitivity or a decrease in the threshold of the peripheral chemoreflex ventilatory response to carbon dioxide. Employing a modified rebreathing technique to assess chemoreflex function, we found evidence of the latter in nine healthy volunteers (six male, three female). Testing consisted of pairs of isoxic rebreathing tests at high and low levels of oxygen, performed before, immediately after and 1 h after a 3 h isocapnic hypoxic exposure. No parameters changed significantly in the high-oxygen rebreathing tests. In the low-oxygen rebreathing tests there were no changes in non chemoreflex ventilatory drives, or in the sensitivity to carbon dioxide, but the carbon dioxide response threshold decreased (approximately 1.5 mmHg) immediately after exposure, and the decrease persisted for 1 h (one-way repeated-measures ANOVA; P < 0.05). We repeated the protocol in five of the original nine volunteers, but this time exposing them to isocapnic normoxia. No trends or significant changes were observed in any of the rebreathing test parameters. These findings demonstrate that in the earliest stages of acclimatisation, there is a decrease in the threshold of the peripheral chemoreflex response to carbon dioxide, which persists for at least 1 h after the return to normoxia. We suggest that ventilatory acclimatisation to hypoxia results from this decreased threshold, reflecting an increase in the activity of the peripheral chemoreflex. PMID- 12562970 TI - Position and velocity responses to galvanic vestibular stimulation in human subjects during standing. AB - Galvanic vestibular stimulation (GVS) in animals modulates the firing of otolith and semicircular canal afferents alike. Here, we look for postural responses evoked by GVS from the otolith organs and semicircular canals. To minimise the modifying effects of somatosensory input on the response, low-intensity (0.3-0.5 mA) GVS was applied for 8 s while subjects stood on foam rubber with the feet together and strapped to the floor. The response had three phases: (i) a rapid movement during the first second, (ii) a slower movement that persisted throughout the stimulus, and (iii) a rapid partial return movement after GVS stopped. The three movement velocities were significantly different. The GVS response therefore appears to be the sum of a step response that returns to the starting point when the stimulus stops, and a constant-velocity ramp response for the duration of the stimulus without a return movement. Subjects' responses differed in size and profile, some with the step or ramp responses almost exclusively but most with a combination of both. The 'step-plus-ramp' model was tested by comparing the three velocities. If the responses add, the initial velocity should not be different from the sum of the velocities during the ramp only period and the step-only period at offset. ANOVA and pairwise comparisons confirmed this. It is concluded that postural responses to GVS arise through stimulation of both otolith and canal afferents. PMID- 12562972 TI - Preventive cardiology: talking it through. PMID- 12562971 TI - Somatic exocytosis of serotonin mediated by L-type calcium channels in cultured leech neurones. AB - We studied somatic exocytosis of serotonin and its mediation by L-type calcium (Ca2+) channels in cultured Retzius neurones of the leech. Exocytosis was induced by trains of impulses at different frequencies or by depolarisation with 40 mM potassium (K+), and was quantified by use of the fluorescent dye FM 1-43. Stimulation increased the membrane fluorescence and produced a pattern of FM 1-43 fluorescent spots of 1.28 +/- 0.01 microm in diameter, provided that Ca2+ was present in the bathing fluid. Individual spots lost their stain during depolarisation with 40 mM K+. Electron micrographs showed clusters of dense core vesicles, some of which were in contact with the cell membrane. Presynaptic structures with clear vesicles were absent from the soma. The number of fluorescent spots per soma, but not their diameter or their fluorescence intensity, depended on the frequency of stimulation. Trains at 1 Hz produced 19.5 +/- 5 spots per soma, 77.9 +/- 13.9 spots per soma were produced at 10 Hz and 91.5 +/- 16.9 spots per soma at 20 Hz. Staining patterns were similar for neurones in culture and in situ. In the presence of the L-type Ca2+ channel blocker nimodipine (10 microM), a 20 Hz train produced only 22.9 +/- 6.4 spots per soma, representing a 75 % reduction compared to control cells (P < 0.05). Subsequent incubation with 10 mM caffeine to induce Ca2+ release from intracellular stores increased the number of spots to 73.22 +/- 12.5. Blockers of N-, P-, Q- or invertebrate Ca2+ channels did not affect somatic exocytosis. Our results suggest that somatic exocytosis by neurones shares common mechanisms with excitable endocrine cells. PMID- 12562973 TI - Fast-track surgical referral in a population displaced by war and conflict. AB - After the 1988-1994 conflict between Armenia and Azerbaijan, fought over the territory of Nagorno-Karabakh, large numbers of people were resettled in camps in southern Azerbaijan. Healthcare in the camps was generally good but there was no access to hospitals. The Leonard Cheshire Centre of Conflict Recovery (LCC) organized a 'fast-track' system of surgical care in the southern camps by securing the help of still-functioning hospitals in the distant capital, Baku. Regular clinics were held in the camps for visiting specialists; and, by arrangement with the Government of Azerbaijan and various non-governmental organizations, treatment was offered to those who fell within strict selection criteria. After a pilot study yielded clear benefits, the scheme was transferred to a local non-governmental organization, which successfully operated an expanded version. The hidden cost of war often includes the neglect of chronic medical conditions that require secondary and tertiary care. The 'fast-track' system illustrates the potential of existing facilities to meet these needs at modest cost, given sufficient support. PMID- 12562974 TI - Homeopathic arnica for prevention of pain and bruising: randomized placebo controlled trial in hand surgery. AB - Homeopathic arnica is widely believed to control bruising, reduce swelling and promote recovery after local trauma; many patients therefore take it perioperatively. To determine whether this treatment has any effect, we conducted a double-blind, placebo-controlled, randomized trial with three parallel arms. 64 adults undergoing elective surgery for carpal tunnel syndrome were randomized to take three tablets daily of homeopathic arnica 30C or 6C or placebo for seven days before surgery and fourteen days after surgery. Primary outcome measures were pain (short form McGill Pain Questionnaire) and bruising (colour separation analysis) at four days after surgery. Secondary outcome measures were swelling (wrist circumference) and use of analgesic medication (patient diary). 62 patients could be included in the intention-to-treat analysis. There were no group differences on the primary outcome measures of pain (P=0.79) and bruising (P=0.45) at day four. Swelling and use of analgesic medication also did not differ between arnica and placebo groups. Adverse events were reported by 2 patients in the arnica 6C group, 3 in the placebo group and 4 in the arnica 30C group. The results of this trial do not suggest that homeopathic arnica has an advantage over placebo in reducing postoperative pain, bruising and swelling in patients undergoing elective hand surgery. PMID- 12562975 TI - Logistic issues and potential prescribing costs associated with use of neuraminidase inhibitors for the treatment of influenza in primary care. AB - In the UK, the National Institute for Clinical Excellence has recommended the use of neuraminidase inhibitors for elderly and at-risk patients who present with influenza-like illness within 36 hours of symptom onset. However, few data exist to enable primary care trusts to evaluate the logistics and costs of prescribing. We sought to determine, during a confirmed influenza outbreak, the proportion of eligible patients who currently present in time to benefit from treatment with a neuraminidase inhibitor, and to develop the findings into a model for evaluating potential prescribing costs. Within a single primary care group, demographic and co-morbidity data were collected on all patients consulting their general practitioner or attending an out-of-hours centre with influenza-like illness during the outbreak period. A typical primary care trust serving 100 000 patients might expect to prescribe a neuraminidase inhibitor to 140 eligible at-risk patients in a season of low influenza activity, rising to 300 in a large epidemic. At-risk patients were more likely than non-at-risk patients to consult within 36 hours of the onset of symptoms. However, only 20% of such patients, rising to 47% in out-of-hours centres, consulted in time to benefit from treatment. The low proportion of elderly and at-risk patients who consult their general practitioner in time to benefit from treatment with a neuraminidase inhibitor emphasizes the overriding importance of annual vaccination in these groups. If the full benefits of neuraminidase inhibitors are to be realized, access to treatment for eligible patients must be improved. PMID- 12562976 TI - A hypothesis to explain how the sensory cortices respond in the appropriate sensory mode. AB - How does an area of sensory cortex recognize the specific nature of the sensory mode of the stimulus that arrives from the peripheral sensory receptor, when nerve impulses are only all-or-nothing action potentials? Work in animals has shown that an area of sensory cortex can learn in which mode to respond. A period of cortical learning is required for phantom limb phenomena to develop, and for the ocular blind to dream in the visual mode. Arguing from these facts I develop the hypothesis that within the sensory cortices there are neurons that learn by neurotropic factor transport from their sensory receptors to function as surrogates for those receptors, thus enabling sensory cortical response to be modally specific. PMID- 12562977 TI - The doctor, the patient and the world-wide web: how the internet is changing healthcare. PMID- 12562978 TI - Chronic headaches due to occult hydrocephalus. PMID- 12562979 TI - Apparent cellulitis with a prolonged APTT. Activated partial thromboplastin time. PMID- 12562980 TI - Acute scrotum after myocardial infarction. PMID- 12562981 TI - Cough and tachypnoea in a toddler. PMID- 12562982 TI - The components of consultation. PMID- 12562983 TI - Gut feelings: a psychologist's 20-year journey with paediatric surgeons. PMID- 12562984 TI - What protection for the unborn child of a psychologically vulnerable adult? PMID- 12562985 TI - Stonehenge: a view from medicine. PMID- 12562986 TI - Shaken baby syndrome. PMID- 12562987 TI - Pooled waiting lists. PMID- 12562988 TI - Capacity and consent. PMID- 12562989 TI - The purpose of RITA. Record of in-training assessment. PMID- 12562990 TI - Supporting individuals with disabling multiple sclerosis. PMID- 12562991 TI - Background and tandem-pore potassium channels in magnocellular neurosecretory cells of the rat supraoptic nucleus. AB - Magnocellular neurosecretory cells (MNCs) were isolated from the supraoptic nucleus of rat hypothalamus, and properties of K(+) channels that may regulate the resting membrane potential and the excitability of MNCs were studied. MNCs showed large transient outward currents, typical of vasopressin- and oxytocin releasing neurons. K(+) channels in MNCs were identified by recording K(+) channels that were open at rest in cell-attached and inside-out patches in symmetrical 150 mM KCl. Eight different K(+) channels were identified and could be distinguished unambiguously by their single-channel kinetics and voltage dependent rectification. Two K(+) channels could be considered functional correlates of TASK-1 and TASK-3, as judged by their single-channel kinetics and high sensitivity to pH(o). Three K(+) channels showed properties similar to TREK type tandem-pore K(+) channels (TREK-1, TREK-2 and a novel TREK), as judged by their activation by membrane stretch, intracellular acidosis and arachidonic acid. One K(+) channel was activated by application of pressure, arachidonic acid and alkaline pH(i), and showed single-channel kinetics indistinguishable from those of TRAAK. One K(+) channel showed strong inward rectification and single channel conductance similar to those of a classical inward rectifier, IRK3. Finally, a K(+) channel whose cloned counterpart has not yet been identified was highly sensitive to extracellular pH near the physiological range similar to those of TASK channels, and was the most active among all K(+) channels. Our results show that in MNCs at rest, eight different types of K(+) channels can be found and six of them belong to the tandem-pore K(+) channel family. Various physiological and pathophysiological conditions may modulate these K(+) channels and regulate the excitability of MNCs. PMID- 12562992 TI - Low doses of ethanol and a neuroactive steroid positively interact to modulate rat GABA(A) receptor function. AB - Fast inhibitory responses in the central nervous system are mediated by the GABA(A) receptor. The activation and function of the GABA(A) receptor can be modulated by a variety of compounds including benzodiazepines, barbiturates and neuroactive steroids. Modulation of the GABA(A) receptor function by ethanol has been observed in some but not all studies. We have studied the effect of ethanol at concentrations corresponding to light intoxication on the function of the recombinant GABA(A) receptor containing alpha1beta2gamma2 subunits. The experiments were performed both in the absence and presence of low, subthreshold concentrations of a neuroactive steroid. The results demonstrate that, in the presence of the steroid, 0.05 % (9 mM) ethanol potentiates the GABA(A) receptor function by increasing the channel mean open duration. No effect was observed on the channel closed time durations. The data suggest that ethanol influences channel closing with no effect on the affinity of the receptor for GABA or the channel opening rate constant. PMID- 12562993 TI - Temperature and redox state dependence of native Kv2.1 currents in rat pancreatic beta-cells. AB - In pancreatic beta-cells, voltage-dependent K(+) (Kv) channels repolarise glucose stimulated action potentials. Kv channels are therefore negative regulators of Ca(2+) entry and insulin secretion. We have recently demonstrated that Kv2.1 mediates the majority of beta-cell voltage-dependent outward K(+) current and now investigate the function of native beta-cell Kv2.1 channels at near-physiological temperatures (32-35 degrees C). While beta-cell voltage-dependent outward K(+) currents inactivated little at room temperature, both fast-inactivation (111.5 +/ 14.3 ms) and slow-inactivation (1.21 +/- 0.12 s) was observed at 32-35 degrees C. Kv2.1 mediates the fast-inactivating current observed at 32-35 degrees C, since it could be selectively ablated by expression of a dominant-negative Kv2.1 construct (Kv2.1N). The surprising ability of Kv2.1N to selectively remove the fast-inactivating component, together with its sensitivity to tetraethylammonium (TEA), demonstrate that this component is not mediated by the classically fast inactivating and TEA-resistant channels such as Kv1.4 and 4.2. Increasing the intracellular redox state by elevating the cytosolic NADPH/NADP(+) ratio from 1/10 to 10/1 increased the rates of both fast- and slow-inactivation. In addition, increasing the intracellular redox state also increased the relative contribution of the fast-inactivation component from 38.8 +/- 2.1 % to 55.9 +/- 1.8 %. The present study suggests that, in beta-cells, Kv2.1 channels mediate a fast-inactivating K(+) current at physiological temperatures and may be regulated by the metabolic generation of NADPH. PMID- 12562994 TI - Group I metabotropic glutamate receptors activate a calcium-sensitive transient receptor potential-like conductance in rat hippocampus. AB - In CA3 pyramidal neurons from organotypic slice cultures, activation of G(q) coupled group I metabotropic glutamate receptors (mGluRs) induces a non-selective cationic conductance that enhances excitability. We have found that this response shares several properties with conductances that are mediated by the transient receptor potential (TRP) family of ion channels, including inhibition by La(3+), 2-aminoethoxydiphenylborane (2APB), cis-N-(2-phenylcyclopentyl)azacyclotridec-1 en-2-amine (MDL 12,330A) and a doubly rectifying current-voltage relationship. Stimulation of mGluR1 and mGluR5 converged to activate the TRP-like conductance in a synergistic manner, and activation of either subtype alone produced only a fraction of the normal response. Activation of the cationic current required elevated intracellular Ca(2+). Chelating intracellular Ca(2+) or blocking Ca(2+) entry through voltage-gated Ca(2+) channels attenuated responses to the activation of mGluRs. Conversely, raising intracellular Ca(2+) potentiated mGluR activation of the TRP-like conductance. Under control conditions, blocking G protein activation using intracellular GDPbetaS with or without N-(2, 6 dimethylphenylcarbamoylmethyl) triethylammonium chloride (QX-314) prevented mGluR mediated activation of the TRP-like conductance. Following G protein blockade, however, the coupling between mGluRs 1 and/or 5 and the TRP-like conductance was rescued by increasing intracellular Ca(2+). This suggests that a G protein independent signalling pathway is also activated by group I mGluRs. Such a pathway may represent an alternative transduction mechanism to maintain metabotropic responses under conditions where G proteins are functionally uncoupled from their cognate receptors. PMID- 12562995 TI - Mossy fibre synaptic NMDA receptors trigger non-Hebbian long-term potentiation at entorhino-CA3 synapses in the rat. AB - Hippocampal CA3 pyramidal cells receive two independent afferents from the enthorinal cortex, i.e. a direct input via the temporoammonic pathway (TA, perforant path) and an indirect input via the mossy fibres (MF) of dentate granule cells. In spite of past suggestions that the TA is assigned an important role in exciting the pyramidal cells, little is known about their physiological properties. By surgically making an incision through the sulcus hippocampi and a small part of the dentate molecular layer, we succeeded in isolating TA-mediated monosynaptic responses in CA3 stratum lacunosum-moleculare. The TA-CA3 synaptic transmission was completely blocked by a combination of D,L-2-amino-5 phosphonopentanoic acid (AP5) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), NMDA and non-NMDA receptor antagonists, respectively, and displayed paired-pulse facilitation and NMDA receptor-dependent long-term potentiation, which are all typical of glutamatergic synapses. We next addressed the heterosynaptic interaction between TA-CA3 and MF-CA3 synapses. The TA-CA3 transmission was partially attenuated by single-pulse MF pre-stimulation at inter-pulse intervals of up to 70 ms. However, surprisingly, burst stimulation of the MF alone induced long-lasting facilitation of TA-CA3 synaptic efficacy. This non-Hebbian form of synaptic plasticity was efficiently prevented by local application of AP5 into the MF synapse-rich area. Therefore, MF-activated NMDA receptors are responsible for the heterosynaptic modification of TA-CA3 transmission, and thereby, the history of MF activity may be etched into TA-CA3 synaptic strength. Our findings predict a novel form of spatiotemporal information processing in the hippocampus, i.e. a use-dependent intersynaptic memory transfer. PMID- 12562996 TI - Orthologous myosin isoforms and scaling of shortening velocity with body size in mouse, rat, rabbit and human muscles. AB - Maximum shortening velocity (V(0)) was determined in single fibres dissected from hind limb skeletal muscles of rabbit and mouse and classified according to their myosin heavy chain (MHC) isoform composition. The values for rabbit and mouse V(0) were compared with the values previously obtained in man and rat under identical experimental conditions. Significant differences in V(0) were found between fibres containing corresponding myosin isoforms in different species: as a general rule for each isoform V(0) decreased with body mass. Myosin isoform distributions of soleus and tibialis anterior were analysed in mouse, rat, rabbit and man: the proportion of slow myosin generally increased with increasing body size. The diversity between V(0) of corresponding myosin isoforms and the different myosin isoform composition of corresponding muscles determine the scaling of shortening velocity of whole muscles with body size, which is essential for optimisation of locomotion. The speed of actin translocation (V(f)) in in vitro motility assay was determined with myosins extracted from single muscle fibres of all four species: significant differences were found between myosin isoforms in each species and between corresponding myosin isoforms in different species. The values of V(0) and V(f) determined for each myosin isoform were significantly correlated, strongly supporting the view that the myosin isoform expressed is the major determinant of maximum shortening velocity in muscle fibres. PMID- 12562997 TI - Glycinergic mIPSCs in mouse and rat brainstem auditory nuclei: modulation by ruthenium red and the role of calcium stores. AB - Spontaneous miniature inhibitory postsynaptic currents (mIPSCs) recorded in central neurons are usually highly variable in amplitude due to many factors such as intrinsic postsynaptic channel fluctuations at each release site, site-to-site variability between release sites, electrotonic attenuation due to variable dendritic locations of synapses, and the possibility of synchronous multivesicular release. A detailed knowledge of these factors is essential for the interpretation of mIPSC amplitude distributions and mean quantal size. We have studied glycinergic mIPSCs in two auditory brainstem nuclei, the rat anteroventral cochlear nucleus (AVCN) and the mouse medial nucleus of the trapezoid body (MNTB). Our previous results have demonstrated the location of glycinergic synapses on these neurons to be somatic, thus avoiding electrotonic complications. Spontaneous glycinergic mIPSCs were recorded from AVCN and MNTB neurons in brainstem slices, in the presence of TTX to block action potentials, and 6-cyano-7-nitroquinoxaline-2, 3-dione, (+/-)-2-amino-5-phosphonopentanoic acid and bicuculline to block glutamatergic and GABAergic synaptic currents. Ruthenium red (RuR), which was used to increase the frequency of mIPSCs, significantly changed the shape of most (90 %) mIPSC amplitude distributions by increasing the proportion of large-amplitude mIPSCs. The possibility was investigated (following previous evidence at GABAergic synapses) that large amplitude glycinergic mIPSCs are due to synchronous multivesicular release initiated by presynaptic calcium sparks from ryanodine-sensitive calcium stores. Interval analysis of mIPSCs indicated that the number of potentially undetected (asynchrony < 0.5 ms) multivesicular mIPSCs was low in comparison with the number of large-amplitude mIPSCs. Ryanodine, thapsigargin and calcium-free perfusate did not reduce the frequency of large-amplitude mIPSCs (> 150 pA), arguing against a significant role for presynaptic calcium stores. Our results support previous evidence suggesting that RuR increases miniature postsynaptic current (mSC) frequency by a mechanism that does not involve presynaptic calcium stores. Our results also indicate that at glycinergic synapses in the AVCN and MNTB, site-to site variability in mIPSC amplitude, rather than multivesicular release, is a major factor underlying the large range of amplitudes of glycinergic mIPSCs. PMID- 12562998 TI - Modulation of inhibitory autapses and synapses on rat CA1 interneurones by GABA(A) receptor ligands. AB - To determine whether autaptic inhibition plays a functional role in the adult hippocampus, the action potential afterhyperpolarisations (spike AHPs) of CA1 interneurones were investigated in 25 basket, three bistratified and eight axo axonic cells. The spike AHPs showed two minima in all regular-spiking (5), burst firing (3) and in many fast-spiking cells (17:28). The fast component had a time to-peak (TTP) of 1.2 +/- 0.5 ms, the slower TTP was very variable (range of 3.3 103 ms). The AHP width at half-amplitude (HW) was 12.5 +/- 5.7 ms in fast spiking, 29.3 +/- 18 ms in regular-spiking and 99.7 +/- 42 ms in burst-firing cells. Axo-axonic cells never establish autapses, and the fast-spiking variety showed narrow (HW: 3.9 +/- 0.7 ms) spike AHPs with only one AHP minimum (TTP: 0.9 +/- 0.1 ms). When challenged with GABA(A) receptor modulators, spike AHPs in basket and bistratified cells were enhanced by zolpidem (HW by 18.4 +/- 6.2 % in 10:15 cells tested), diazepam (45.2 +/- 0.5 %, 6:7), etomidate (43.9 +/- 36 %, 6:8) and pentobarbitone sodium (41 %, 1:1), and were depressed by bicuculline ( 41 +/- 5.7 %, 5:8) and picrotoxin (-54 %, 1:1), and the enhancement produced by zolpidem was reduced by flumazenil (-31 +/- 13 %, relative to the AHP HW during exposure to zolpidem, 3:4). Neuronal excitability was modulated in parallel. The spike AHPs of three axo-axonic cells tested showed no sensitivity to etomidate, pentobarbitone or diazepam. Interneurone-to-interneurone inhibitory postsynaptic potentials (IPSPs), studied with dual intracellular recordings, had time courses resembling those of the spike AHPs. The IPSP HW was 13.4 +/- 2.8 ms in fast spiking (n = 16) and 28.7 +/- 5.8 ms in regular-spiking/burst-firing cells (n = 6), and the benzodiazepine1-selective modulator zolpidem strongly enhanced these IPSPs (45 +/- 28 %, n = 5). Interneurones with spike AHPs affected by the GABA(A) receptor ligands exhibited 3.8 +/- 1.9 close autaptic appositions. In three basket cells studied at the ultrastructural level 6 of 6, 1 of 2 and 1 of 2 close appositions were confirmed as autapses. Therefore, in the hippocampus autaptic connections contribute to spike AHPs in many interneurones. These autapses influence neuronal firing and responses to GABA(A) receptor ligands. PMID- 12562999 TI - Glutamate modulation of GABA transport in retinal horizontal cells of the skate. AB - Transport of the amino acid GABA into neurons and glia plays a key role in regulating the effects of GABA in the vertebrate retina. We have examined the modulation of GABA-elicited transport currents of retinal horizontal cells by glutamate, the likely neurotransmitter of vertebrate photoreceptors. Enzymatically isolated external horizontal cells of skate were examined using whole-cell voltage-clamp techniques. GABA (1 mM ) elicited an inward current that was completely suppressed by the GABA transport inhibitors tiagabine (10 microM) and SKF89976-A (100 microM), but was unaffected by 100 microM picrotoxin. Prior application of 100 microM glutamate significantly reduced the GABA-elicited current. Glutamate depressed the GABA dose-response curve without shifting the curve laterally or altering the voltage dependence of the current. The ionotropic glutamate receptor agonists kainate and AMPA also reduced the GABA-elicited current, and the effects of glutamate and kainate were abolished by the ionotropic glutamate receptor antagonist 6-cyano-7-nitroquinoxaline. NMDA neither elicited a current nor modified the GABA-induced current, and metabotropic glutamate analogues were also without effect. Inhibition of the GABA-elicited current by glutamate and kainate was reduced when extracellular calcium was removed and when recording pipettes contained high concentrations of the calcium chelator BAPTA. Caffeine (5 mM) and thapsigargin (2 nM), agents known to alter intracellular calcium levels, also reduced the GABA-elicited current, but increases in calcium induced by depolarization alone did not. Our data suggest that glutamate regulates GABA transport in retinal horizontal cells through a calcium-dependent process, and imply a close physical relationship between calcium-permeable glutamate receptors and GABA transporters in these cells. PMID- 12563000 TI - Oscillations in ciliary beat frequency and intracellular calcium concentration in rabbit tracheal epithelial cells induced by ATP. AB - To investigate how Ca(2+) regulates airway ciliary activity, changes in ciliary beat frequency (CBF) and intracellular calcium concentration ([Ca(2+)](i)) of rabbit tracheal ciliated cells, in response to ATP, were simultaneously quantified with high-speed phase-contrast and fast fluorescence imaging. [ATP] 1 but < 16 microM induced a similar increase in [Ca(2+)](i) and CBF but this was followed by oscillations in CBF and [Ca(2+)](i). The minimum CBF of the oscillations in CBF remained elevated above the basal rate while the minimum concentration of the [Ca(2+)](i) oscillations returned to the basal level. The minimum and maximum CBF of the oscillations in CBF were independent of the [ATP], whereas the frequency of the oscillations in CBF was dependent on the [ATP]. Similar oscillations in CBF and [Ca(2+)](i) were induced by ATP- gamma -S. Although ADP, AMP and adenosine induced a Ca(2+)-independent increase in CBF, neither ATP nor ATP- gamma -S induced an increase in CBF when the Ca(2+) increases were abolished by 20 microM BAPTA AM, a result suggesting that ATP hydrolysis was minimal. [ATP] >or=16 microM induced a sustained elevation in CBF and only a temporary, non-oscillating increase in [Ca(2+)](i). A similar response was induced by thapsigargin (2 microM). Flash photolysis of caged Ca(2+) (NP-EGTA) produced both transient and prolonged increases in [Ca(2+)](i) which were accompanied by transient and sustained increases in CBF, respectively. From these results, we propose that CBF can be increased by a direct Ca(2+) -dependent mechanism that generates the rapid increases in CBF associated with the oscillations or by an indirect Ca(2+)-dependent mechanism that is responsible for the sustained minimum increase in CBF. PMID- 12563001 TI - Involvement of intramuscular interstitial cells in nitrergic inhibition in the mouse gastric antrum. AB - Intracellular recordings were made from isolated bundles of the circular muscle layer of mouse gastric antrum and the responses evoked by stimulating intrinsic nerve fibres were examined. Transmural nerve stimulation evoked a fast inhibitory junction potential (fast-IJP) which was followed initially by a smaller amplitude long lasting inhibitory junction potential (slow-IJP) and a period of excitation. The excitatory component of the response was abolished by atropine, suggesting that it resulted from the release of acetylcholine and activation of muscarinic receptors. Fast-IJPs were selectively reduced in amplitude by apamin and slow IJPs were abolished by N(omega)-nitro-L-arginine. Slow-IJPs were associated with a drop in membrane noise, suggesting that inhibition resulted from a reduced discharge of unitary potentials by intramuscular interstitial cells of Cajal (ICC(IM)). The chloride channel blocker, anthracene-9-carboxylic acid, reduced the discharge of membrane noise in a manner similar to that detected during the slow-IJP. When recordings were made from the antrum of W/W(V) mice, which lack ICC(IM), the cholinergic and nitrergic components were absent, with only fast IJPs being detected. The observations suggest that neurally released nitric oxide selectively targets ICC(IM) causing a hyperpolarization by suppressing the discharge of unitary potentials. PMID- 12563002 TI - Ubiquitin-proteasome-dependent muscle proteolysis responds slowly to insulin release and refeeding in starved rats. AB - The central role of the ubiquitin-proteasome system in the loss of skeletal muscle protein in many wasting conditions has been well established. However, it is unclear what factors are responsible for the suppression of this system during periods of protein gain. Thus, the aim of these studies was to examine the short term effects of insulin release and nutrients on skeletal muscle protein turnover in young rats starved for 48 h, and then infused intravenously with amino acids (AA), or fed an oral diet. Forty-eight hours of starvation (i.e. prolonged starvation in young rats) decreased muscle protein synthesis and increased proteasome-dependent proteolysis. Four-hour AA infusion and 4 h of refeeding increased plasma insulin release and AA concentrations, and stimulated muscle protein synthesis, but had no effect on either total or proteasome-dependent proteolysis, despite decreased plasma corticosterone concentrations. Both muscle proteasome-dependent proteolysis and the rate of ubiquitination of muscle proteins were not suppressed until 10 h of refeeding. The temporal response of these two measurements correlated with the normalised expression of the 14-kDa E2 (a critical enzyme in substrate ubiquitination in muscle) and the expression of the MSS1 subunit of the 19S regulatory complex of the 26S proteasome. In contrast, the starvation-induced increase in mRNA levels for 20S proteasome subunits was normalised by refeeding within 24 h in muscle, and 6 h in jejunum, respectively. In conclusion, unlike protein synthesis, skeletal muscle proteasome dependent proteolysis is not acutely responsive in vivo to insulin, AA, and/or nutrient intake in refed starved rats. This suggests that distinct and perhaps independent mechanisms are responsible for the nutrient-dependent regulation of protein synthesis and ubiquitin-proteasome-dependent proteolysis following a prolonged period of catabolism. Furthermore, factors other than the expression of ubiquitin-proteasome pathway components appear to be responsible for the suppression of skeletal muscle proteasome-dependent proteolysis by nutrition. PMID- 12563003 TI - Evolution of regional performance after an acute anterior myocardial infarction in humans using magnetic resonance tagging. AB - Regional remodelling after a left ventricular myocardial infarction is the first step in a cascade that may lead to heart failure and death. To understand better the mechanisms underlying this process, it is important to study not only the evolution in local deformation parameters but also the corresponding loading conditions. Using magnetic resonance (MR) myocardial tagging, we measured the regional contribution to ejection (regional ejection fraction) and loading (systolic blood pressure x radius of curvature (mean of short and long axes)/wall thickness) in 32 regions throughout the left ventricle (LV) in patients 1 week (1W) and 3 months (3M) after a first anterior infarction. Using positron emission tomography (PET), the LV was divided into infarct, adjacent and remote regions. In the remote regions the average deformation decreased between 1W and 3M (from 59.3 +/- 5.6 to 57.9 +/- 6.7 %, P < 0.05) due to an increase in loading conditions only (from 730 +/- 290 to 837 +/- 299 mmHg, P < 0.05). In the adjacent myocardium, no change in function was observed (49.0 +/- 10.8 to 49.0 +/- 6.5 %, P = n.s.), although loading increased (806 +/- 297 to 978 +/- 287 mmHg, P < 0.05). In the infarct region only, an increase in deformation was seen (30.7 +/- 14.2 to 37 +/- 6.9 %, P < 0.05), together with a higher loading level (1229 +/- 422 to 1466 +/- 284 mmHg, P < 0.05), which indicates a true improvement in function. The fact that MR tagging can identify both regional deformation and loading permits us to differentiate between changes due to alterations in regional loading conditions and true changes in function. After an acute myocardial infarction (MI), an improvement can be observed in the deformation loading relation in the adjacent and infarct regions, but the improvement is mainly in the infarct region. Using this technique, types of intervention leading to even more functional gain could be evaluated. PMID- 12563004 TI - Functional analysis of neurotransmission at beta2-laminin deficient terminals. AB - beta2-Laminin is important for the formation of neuromuscular junctions in vertebrates. Previously, we have inactivated the gene that encodes for beta2 laminin in mice and observed predominantly prejunctional structural defects. In this study, we have used both intra- and extracellular recording methods to investigate evoked neurotransmission in beta2-laminin-deficient mice, from postnatal day 8 (P8) through to day 18 (P18). Our results confirmed that there was a decrease in the frequency of spontaneous release, but no change in the postjunctional response to such release. Analysis of evoked neurotransmission showed an increase in the frequency of stimuli that failed to elicit an evoked postjunctional response in the mutants compared to litter mate controls, resulting in a 50 % reduction in mean quantal content at mutant terminals. Compared to littermate controls, beta2-laminin-deficient terminals showed greater synaptic depression when subjected to high frequency stimulation. Furthermore, the paired pulse ratio of the first two stimuli was significantly lower in beta2 laminin mutant terminals. Statistical analysis of the binomial parameters of release showed that the decrease in quantal content was due to a decrease in the number of release sites without any significant change in the average probability of release. This suggestion was supported by the observation of fewer synaptic vesicle protein 2 (SV2)-positive varicosities in beta2-laminin-deficient terminals and by ultrastructural observations showing smaller terminal profiles and increased Schwann cell invasion in beta2-laminin mutants; the differences between beta2-laminin mutants and wild-type mice were the same at both P8 and P18. From these results we conclude that beta2-laminin plays a role in the early structural development of the neuromuscular junction. We also suggest that transmitter release activity may act as a deterrent to Schwann cell invasion in the absence of beta2-laminin. PMID- 12563005 TI - Strong, reliable and precise synaptic connections between thalamic relay cells and neurones of the nucleus reticularis in juvenile rats. AB - The thalamic reticular nucleus (nRT) is composed entirely of GABAergic inhibitory neurones that receive input from pyramidal cortical neurones and excitatory relay cells of the ventrobasal complex of the thalamus (VB). It plays a major role in the synchrony of thalamic networks, yet the synaptic connections it receives from VB cells have never been fully physiologically characterised. Here, whole-cell current-clamp recordings were obtained from 22 synaptically connected VB-nRT cell pairs in slices of juvenile (P14-20) rats. At 34-36 degrees C, single presynaptic APs evoked unitary EPSPs in nRT cells with a peak amplitude of 7.4 +/- 1.5 mV (mean +/- S.E.M.) and a decay time constant of 15.1 +/- 0.9 ms. Only four out of 22 pairs showed transmission failures at a mean rate of 6.8 +/- 1.1 %. An NMDA receptor (NMDAR)-mediated component was significant at rest and subsequent EPSPs in a train were depressed. Only one out of 14 pairs tested was reciprocally connected; the observed IPSPs in the VB cell had a peak amplitude of 0.8 mV and were completely abolished in the presence of 10 microM bicuculline. Thus, synaptic connections from VB cells to nRT neurones are mainly 'drivers', while a small subset of cells form closed disynaptic loops. PMID- 12563006 TI - The effect of chronic skeletal muscle stimulation on capillary growth in the rat: are sensory nerve fibres involved? AB - Indirect chronic electrical stimulation of skeletal muscle activates not only efferent but also afferent nerve fibres. To investigate effects specific to this on capillary growth, one of the earliest changes, cell proliferation and capillary ultrastructure were studied in ankle flexors of rats with and without deafferentation of the stimulated side. Two weeks after preganglionic section of dorsal roots L4-L6, the peroneal nerve was stimulated (10 Hz, 8 h day(-1)) for 2 or 7 days. Proliferating nuclei labelled by bromodeoxyuridine or proliferating cell nuclear antigen staining were colocalized to alkaline phosphatase-stained capillaries (Lc) or other interstitial nuclei (Li) in frozen sections of extensor digitorum longus. Capillary fine structure was examined in extensor hallucis proprius by transmission electron microscopy. The stimulation-induced increase in capillary and interstitial proliferation (Lc 9.9 +/- 1.9 %, Li 8.8 +/- 2.1 % vs. Lc 2.6 +/- 0.4 %, Li 1.9 +/- 0.3 % in controls, P < 0.05) was depressed at 2 days by dorsal root section (Lc 4.8 +/- 0.7 %, Li 3.2 +/- 0.9 %, P < 0.05), an effect likely to be mainly on fibroblasts; no depression was seen at 7 days. Dorsal root section reduced stimulation-induced capillary endothelial swelling at both time points. In contralateral muscles of intact rats, stimulation increased interstitial cell proliferation and capillary swelling, both effects being eliminated by dorsal root section. Capillary growth induced by stimulation (24 % increase in capillary : fibre ratio at 7 days) was unaffected by deafferentation. The reduction in capillary ultrastructural changes and interstitial proliferation in both stimulated and contralateral muscles implies that stimulation of afferent fibres leads directly to release of humoral factors and/or activation via dorsal roots of fibres that release humoral substances. Contralateral muscles are an inadequate control for the effects of chronic stimulation in the intact animal. PMID- 12563007 TI - Essential role of rho kinase in the Ca2+ sensitization of prostaglandin F(2alpha) induced contraction of rabbit aortae. AB - Inhibition of dephosphorylation of the 20 kDa myosin light chain (MLC(20)) is an important mechanism for the Ca(2+)-induced sensitization of vascular smooth muscle contraction. We investigated whether this mechanism operates in prostaglandin F(2alpha) (PGF(2alpha))-induced contraction of rabbit aortic smooth muscle and, if so, whether protein kinase C (PKC) or rho-associated kinase (rho kinase) contribute to the inhibition of dephosphorylation. In normal medium, PGF(2alpha) (10 microM) increased the phosphorylation of MLC(20) and developed tension. The rho-kinase inhibitors fasudil and hydroxyfasudil inhibited these changes, despite having no effect on a phorbol-ester-induced MLC(20) phosphorylation. After treatment with verapamil or chelation of external Ca(2+) with EGTA, PGF(2alpha) increased the MLC(20) phosphorylation and tension without an increase in [Ca(2+)](i), all of which were sensitive to fasudil and hydroxyfasudil. ML-9, a MLC kinase inhibitor, quickly reversed the KCl-induced MLC(20) phosphorylation and contraction to the resting level. However, fractions of PGF(2alpha)-induced contraction and MLC(20) phosphorylation were resistant to ML-9 but were sensitive to fasudil. Ro31-8220 (10 microM), a PKC inhibitor, did not affect the phosphorylation of MLC(20) and the tension caused by PGF(2alpha), thus excluding the possibility of the involvement of PKC in the PGF(2alpha) induced MLC(20) phosphorylation. PGF(2alpha) increased phosphorylation at Thr654 of the myosin binding subunit (MBS) of myosin phosphatase, which is a target of rho kinase, and fasudil decreased the phosphorylation. These data suggest that the PGF(2alpha)-induced contraction is accompanied by the inhibition of MLC(20) dephosphorylation through rho kinase-induced MBS phosphorylation, leading to Ca(2+) sensitization of contraction. An actin-associated mechanism may also be involved in the PGF(2alpha)-induced sensitization. PMID- 12563008 TI - Autonomic nervous control of myoepithelial cells and secretion in submandibular gland of anaesthetized dogs. AB - In dog submandibular gland, the activity of myoepithelial cells was assessed by simultaneous measurement of intraductal pressure (P(du)) and subcapsular pressure (P(ca)) using catheter-tip pressure transducers; their resting values were 2.5 +/ 0.21 and 3.0 +/- 0.19 mmHg, respectively (n = 40). Retrograde infusion of saliva (collected from preceding parasympathetic nerve stimulation) increased P(du) (coefficient of 50 mmHg ml(-1) for rates < 1 ml min(-1) and 85 mmHg ml(-1) for higher rates) and P(ca) (coefficient of 0.47 mmHg ml(-1) for all rates). Blood flow changes did not affect P(du) but increased P(ca) (coefficient of 0.04 mmHg ml(-1)). Parasympathetic nerve stimulation increased P(du) but decreased P(ca) abruptly; the response threshold was 0.1 Hz, with maximal responses at 16 Hz. The coefficients for P(du) and P(ca) on salivary secretion to parasympathetic nerve stimulation in glands with spontaneous blood flow (5.3 x 10(-3) and 4.87 x 10(-2) ml min(-1) g(-1) mmHg(-1)) were close to their values in glands with constant flow vascular perfusion (4.9 x 10(-3) and 3.68 x 10(-2) ml min(-1) g(-1) mmHg( 1)). The finding that P(ca) fell despite concomitant increased blood flow suggests contraction of myoepithelial cells. Additional ductal occlusion further increased P(du) and enhanced the fall in P(ca), suggesting that the myoepithelial cells can contract when distended. Atropine blocked salivary secretion and responses of P(du) and P(ca) to parasympathetic nerve stimulation. ACh elicited responses similar to that of parasympathetic nerve stimulation. VIP caused very scanty salivary secretion and gradual slight increases in P(du) and P(ca); the change in P(ca) was abolished in glands with constant-flow vascular perfusion. Hence, contraction of myoepithelial cells to parasympathetic nerve stimulation is via muscarinic receptors. Sympathetic nerve stimulation increased P(du) and decreased P(ca) abruptly; the response threshold was 0.1 Hz, with maximal responses at 16 Hz. The coefficients for P(du) and P(ca) on salivary secretion to sympathetic nerve stimulation in glands with spontaneous blood flow (3.0 x 10(-3) and 3.2 x 10(-3) ml min(-1) g(-1) mmHg(-1)) were similar to their values in glands with constant-flow vascular perfusion (3.2 x 10(-3) and 3.1 x 10(-3) ml min(-1) g(-1) mmHg(-1)). The finding that P(ca) fell even in glands with constant flow vascular perfusion suggests contraction of myoepithelial cells. Superimposed sympathetic nerve stimulation immediately enhanced the pressure changes and secretory response to parasympathetic nerve stimulation, indicating that the two autonomic nerves act synergistically to evoke myoepithelial cell contraction. Phentolamine and prazosin but not propranolol and yohimbine blocked the sympathetic enhancement. The finding that phenylephrine, but not clonidine and isoproterenol, abruptly decreased P(ca) in glands with constant-flow vascular perfusion suggests that the sympathetic activation of myoepithelial cells is via the alpha(1)-adrenoceptors. PMID- 12563009 TI - Exercise induces transient transcriptional activation of the PGC-1alpha gene in human skeletal muscle. AB - Endurance exercise training induces mitochondrial biogenesis in skeletal muscle. The peroxisome proliferator activated receptor co-activator 1alpha (PGC-1alpha) has recently been identified as a nuclear factor critical for coordinating the activation of genes required for mitochondrial biogenesis in cell culture and rodent skeletal muscle. To determine whether PGC-1alpha transcription is regulated by acute exercise and exercise training in human skeletal muscle, seven male subjects performed 4 weeks of one-legged knee extensor exercise training. At the end of training, subjects completed 3 h of two-legged knee extensor exercise. Biopsies were obtained from the vastus lateralis muscle of both the untrained and trained legs before exercise and after 0, 2, 6 and 24 h of recovery. Time to exhaustion (2 min maximum resistance), as well as hexokinase II (HKII), citrate synthase and 3-hydroxyacyl-CoA dehydrogenase mRNA, were higher in the trained than the untrained leg prior to exercise. Exercise induced a marked transient increase (P < 0.05) in PGC-1alpha transcription (10- to > 40-fold) and mRNA content (7- to 10-fold), peaking within 2 h after exercise. Activation of PGC 1alpha was greater in the trained leg despite the lower relative workload. Interestingly, exercise did not affect nuclear respiratory factor 1 (NRF-1) mRNA, a gene induced by PGC-1alpha in cell culture. HKII, mitochondrial transcription factor A, peroxisome proliferator activated receptor alpha, and calcineurin Aalpha and Abeta mRNA were elevated (approximately 2- to 6-fold; P < 0.05) at 6 h of recovery in the untrained leg but did not change in the trained leg. The present data demonstrate that exercise induces a dramatic transient increase in PGC-1alpha transcription and mRNA content in human skeletal muscle. Consistent with its role as a transcriptional coactivator, these findings suggest that PGC 1alpha may coordinate the activation of metabolic genes in human muscle in response to exercise. PMID- 12563010 TI - PSD-95 regulates synaptic transmission and plasticity in rat cerebral cortex. AB - PSD-95 is one of the most abundant proteins found in the postsynaptic density of excitatory synapses. However, the precise functional role played by PSD-95 in regulating synaptic transmission and plasticity remains undefined. To address this issue, we have overexpressed PSD-95 in cortical pyramidal neurons in organotypic brain slices using particle-mediated gene transfer and assessed the consequences on synaptic transmission and plasticity. The AMPA receptor/NMDA receptor (AMPAR/NMDAR) ratio of evoked EPSCs recorded at +40 mV was greater in PSD-95-transfected pyramidal neurons than in controls. This difference could not be accounted for by a change in rectification of AMPAR-mediated synaptic currents since the current-voltage curves obtained in controls and in PSD-95-transfected neurons were indistinguishable. However, the amplitude of AMPAR-mediated evoked EPSCs was larger in PSD-95-transfected neurons compared to matched controls. Paired-pulse ratio analysis suggested that overexpression of PSD-95 did not alter presynaptic release probability. Transfection of PSD-95 was further accompanied by an increase in the frequency, but not amplitude, of AMPAR-mediated mEPSCs. Together, these results indicate that transfection of PSD-95 increased AMPAR mediated synaptic transmission. Furthermore, they suggest that this phenomenon reflects an increased number of synapses expressing AMPARs rather than an increased number or function of these receptors at individual synapses. We tested the consequences of these changes on synaptic plasticity and found that PSD-95 transfection greatly enhanced the probability of observing long-term depression. These results thus identify a physiological role for PSD-95 and demonstrate that this protein can play a decisive role in controlling synaptic strength and activity-dependent synaptic plasticity. PMID- 12563011 TI - Fetal lamb cerebral blood flow (CBF) and oxygen tensions during hypoxia: a comparison of laser Doppler and microsphere measurements of CBF. AB - This study was undertaken to compare microsphere and laser Doppler flowmetry techniques for the measurement of cerebral blood flow, to assess the effect of probe implantation at the tip of the sensing probe and to measure brain tissue P(O2) (tP(O2)) in response to acute hypoxia. Fetal sheep of ~131 days gestation (n = 8) were chronically instrumented with bilateral laser Doppler probes in the parietal cortices and catheters for injection of fluorescent microspheres. Five days after surgery fetuses were subjected to 1 h periods of baseline control breathing, hypoxia and recovery. Microspheres were injected 10 min prior to and 10, 30, 50 and 120 min after initiation of hypoxia. Microspheres were counted in four 12 mm(3) tissue samples from each hemisphere, the tip of the laser Doppler probe being positioned in the centre of one of the cubes. The cube containing the probe tip was also subdivided into 4 mm(3) pieces of tissue. In response to hypoxia, fetal arterial P(O2) declined from 21 +/- 2 to 12 +/- 1 Torr and brain tissue P(O2) fell from 10 +/- 1 to a nadir of 1 +/- 1 Torr. Each method detected a significant increase in CBF that reached a maximum after 30-45 min, although the increase of flow measured by laser Doppler flowmetry was less than that measured by spheres after 10 and 30 min (P < 0.05). Microspheres did not detect altered flow at the probe tip or heterogeneity of flow in surrounding volumes of cortical tissue. In summary, laser Doppler flowmetry is a useful measure of continuous relative changes of CBF in the chronically instrumented fetal sheep. Flow compensations in acute hypoxia are not adequate to sustain O(2) delivery, and other compensations, including reduced metabolic rate, are possible. PMID- 12563012 TI - Phosphorylation of the myosin phosphatase targeting subunit and CPI-17 during Ca2+ sensitization in rabbit smooth muscle. AB - Myosin phosphatase (MLCP) plays a critical regulatory role in the Ca(2+) sensitivity of myosin phosphorylation and smooth muscle contraction. It has been suggested that phosphorylation at Thr(695) of the MLCP regulatory subunit (MYPT1) and at Thr(38) of the MLCP inhibitor protein CPI-17 results in inhibition of MLCP activity. We have previously demonstrated that CPI-17 Thr(38) phosphorylation plays an important role in G-protein-mediated inhibition of MLCP in tonic arterial smooth muscle. Here, we attempted to evaluate the function of MYPT1 in phasic rabbit portal vein (PV) and vas deferens (VD) smooth muscles. Using site- and phospho-specific antibodies, phosphorylation of MYPT1 Thr(695) and CPI-17 Thr(38) was examined along with MYPT1 Thr(850), which is a non-inhibitory Rho kinase site. We found that both CPI-17 Thr(38) and MYPT1 Thr(850) were phosphorylated in response to agonists or GTPgammaS concurrently with contraction and myosin phosphorylation in alpha-toxin-permeabilized PV tissues. In contrast, phosphorylation of MYPT1 Thr(695) did not increase. Comparable results were also obtained in both permeabilized and intact VD. The Rho-kinase inhibitor Y-27632 and the protein kinase C (PKC) inhibitor GF109203X suppressed phosphorylation of MYPT1 Thr(850) and CPI-17 Thr(38), respectively, in intact VD while MYPT1 Thr(695) phosphorylation was insensitive to both inhibitors. These results indicate that phosphorylation of MYPT1 Thr(695) is independent of stimulation of G-proteins, Rho-kinase or PKC. In the phasic PV, phosphorylation of CPI-17 Thr(38) may contribute towards inhibition of MLCP while the phasic visceral VD, which has a low CPI-17 concentration, probably utilizes other Ca(2+) sensitizing mechanisms for inhibiting MLCP besides phosphorylation of MYPT1 and CPI-17. PMID- 12563013 TI - The role of neuropeptide Y in the ovine fetal cardiovascular response to reduced oxygenation. AB - This study investigated the role of neuropeptide Y (NPY) in mediating cardiovascular responses to reduced oxygenation in the late gestation ovine fetus by: (1) comparing the effects on the cardiovascular system of an exogenous infusion of NPY with those elicited by moderate or severe reductions in fetal oxygenation; and (2) determining the effect of fetal I.V. treatment with a selective NPY-Y(1) receptor antagonist on the fetal cardiovascular responses to acute moderate hypoxaemia. Under general anaesthesia, 14 sheep fetuses (0.8-0.9 of gestation) were surgically prepared with vascular and amniotic catheters. In 5 of these fetuses, a Transonic flow probe was also implanted around a femoral artery. Following at least 5 days of recovery, one group of fetuses (n = 9) was subjected to a 30 min treatment period with exogenous NPY (17 microg kg(-1) bolus plus 0.85 microg kg(-1) min(-1) infusion). In this group, fetal blood pressure and heart rate were monitored continuously and the distribution of the fetal combined ventricular output was assessed via injection of radiolabelled microspheres before and during treatment. The second group of fetuses instrumented with the femoral flow probe (n = 5) were subjected to a 3 h experiment consisting of 1 h of normoxia, 1 h of hypoxaemia, and 1 h of recovery during a slow I.V. infusion of vehicle. One or two days later, the acute hypoxaemia protocol was repeated during fetal I.V. treatment with a selective NPY Y(1) receptor antagonist (50 microg kg(-1) bolus + 1.5 microg kg(-1) min(-1) infusion). In these fetuses, fetal arterial blood pressure, heart rate and femoral vascular resistance were recorded continuously. The results show that fetal treatment with exogenous NPY mimics the fetal cardiovascular responses to asphyxia, and that treatment of the sheep fetus with a selective NPY-Y(1) receptor antagonist does not affect the fetal cardiovascular response to acute moderate hypoxaemia. These results support a greater role for NPY in mediating the fetal cardiovascular responses to acute asphyxia than to acute moderate hypoxaemia. PMID- 12563014 TI - Centre and surround responses of marmoset lateral geniculate neurones at different temporal frequencies. AB - The responses of marmoset lateral geniculate neurones to stimuli that were composed of a sinusoidally modulating centre stimulus and a surround that was modulated in counterphase were measured. The size of the stimulus centre was varied. These measurements were repeated at different temporal frequencies between 1 and 30 Hz. The response amplitudes and phases depended in a characteristic manner on the stimulus centre size. The response behaviour could be modelled by assuming Gaussian responsivity profiles of the cells' receptive field (RF) centres and surrounds and a phase delay in the RF surround responses, relative to the centre, enabling the description of RF centre and surround response characteristics. We found that the RF centre-to-surround phase difference increased linearly with increasing temporal frequency, indicating a constant delay difference of about 4.5 to 6 ms. A linear model, including low pass filters, a lead lag stage and a delay, was used to describe the mean RF centre and surround responses. The separate RF centre and surround responses were less band pass than the full receptive field responses of the cells. The linear model provided less satisfactory fits to M-cell responses than to those of P cells, indicating additional nonlinearities. PMID- 12563015 TI - Sympathetic neural overactivity in healthy humans after prolonged exposure to hypobaric hypoxia. AB - Acute exposure to hypoxia causes chemoreflex activation of the sympathetic nervous system. During acclimatization to high altitude hypoxia, arterial oxygen content recovers, but it is unknown to what degree sympathetic activation is maintained or normalized during prolonged exposure to hypoxia. We therefore measured sympathetic nerve activity directly by peroneal microneurography in eight healthy volunteers (24 +/- 2 years of age) after 4 weeks at an altitude of 5260 m (Chacaltaya, Bolivian Andes) and at sea level (Copenhagen). The subjects acclimatized well to altitude, but in every subject sympathetic nerve activity was highly elevated at altitude vs. sea level (48 +/- 5 vs. 16 +/- 3 bursts min( 1), respectively, P < 0.05), coinciding with increased mean arterial blood pressure (87 +/- 3 vs. 77 +/- 2 mmHg, respectively, P < 0.05). To examine the underlying mechanisms, we administered oxygen (to eliminate chemoreflex activation) and saline (to reduce cardiopulmonary baroreflex deactivation). These interventions had minor effects on sympathetic activity (48 +/- 5 vs. 38 +/- 4 bursts min(-1), control vs. oxygen + saline, respectively, P < 0.05). Moreover, sympathetic activity was still markedly elevated (37 +/- 5 bursts min(-1)) when subjects were re-studied under normobaric, normoxic and hypervolaemic conditions 3 days after return to sea level. In conclusion, acclimatization to high altitude hypoxia is accompanied by a striking and long-lasting sympathetic overactivity. Surprisingly, chemoreflex activation by hypoxia and baroreflex deactivation by dehydration together could account for only a small part of this response, leaving the major underlying mechanisms unexplained. PMID- 12563016 TI - The effect of diazepam on motor cortical oscillations and corticomuscular coherence studied in man. AB - EEG recordings from sensorimotor cortex show oscillations around 10 and 20 Hz. These modulate with task performance, and are strongest during periods of steady contraction. The 20 Hz oscillations are coherent with contralateral EMG. Computer modelling suggests that oscillations arising within the cortex may be especially dependent on inhibitory systems. The benzodiazepine diazepam enhances the size of GABA(A) IPSPs; its effects are reversed by the antagonist flumazenil. We tested the effect of these drugs on spectral measures of EEG and EMG, whilst eight healthy human subjects performed a precision grip task containing both holding and movement phases. Either an auxotonic or isometric load was used. EEG changes following electrical stimulation of the contralateral median nerve were also assessed. The EEG power showed similar changes in all task/stimulation protocols used. Power around 20 Hz doubled at the highest dose of diazepam used (5 mg), and returned to control levels following flumazenil. EEG power at 10 Hz was by contrast little altered. The peak frequency of EEG power in both bands was not changed by diazepam. Corticomuscular coherence at ca 20 Hz was reduced following diazepam injection, but the magnitude of this effect was small (mean coherence during steady holding in the auxotonic task was 0.062 in control recordings, 0.051 after 2.5 mg and 5 mg doses of diazepam). These results imply that 20 Hz oscillations in the sensorimotor cortex are at least partially produced by local cortical circuits reliant on GABA(A)-mediated intracortical inhibition, whereas 10 Hz rhythms arise by a different mechanism. Rhythms generated during different tasks, or following nerve stimulation, are likely to arise from similar mechanisms. By examining the formulae used to calculate coherence, we show that if cortical oscillations are simply transmitted to the periphery, corticomuscular coherence should increase in parallel with the ratio of EEG to EMG power. The relative constancy of coherence even when the amplitude of cortical oscillations is perturbed suggests that corticomuscular coherence itself may have a functional role in motor control. PMID- 12563017 TI - Distribution of inspiratory drive to the external intercostal muscles in humans. AB - The external intercostal muscles in humans show marked regional differences in respiratory effect, and this implies that their action on the lung during breathing is primarily determined by the spatial distribution of neural drive among them. To assess this distribution, monopolar electrodes were implanted under ultrasound guidance in different muscle areas in six healthy individuals and electromyographic recordings were made during resting breathing. The muscles in the dorsal portion of the third and fifth interspace showed phasic inspiratory activity with each breath in every subject. However, the muscle in the ventral portion of the third interspace showed inspiratory activity in only three subjects, and the muscle in the dorsal portion of the seventh interspace was almost invariably silent. Also, activity in the ventral portion of the third interspace, when present, and activity in the dorsal portion of the fifth interspace were delayed relative to the onset of activity in the dorsal portion of the third interspace. In addition, the discharge frequency of the motor units identified in the dorsal portion of the third interspace averaged (mean +/- S.E.M.) 11.9 +/- 0.3 Hz and was significantly greater than the discharge frequency of the motor units in both the ventral portion of the third interspace (6.0 +/- 0.5 Hz) and the dorsal portion of the fifth interspace (6.7 +/- 0.4 Hz). The muscle in the dorsal portion of the third interspace started firing simultaneously with the parasternal intercostal in the same interspace, and the discharge frequency of its motor units was even significantly greater (11.4 +/- 0.3 vs. 8.9 +/- 0.2 Hz). These observations indicate that the distribution of neural inspiratory drive to the external intercostals in humans takes place along dorsoventral and rostrocaudal gradients and mirrors the spatial distribution of inspiratory mechanical advantage. PMID- 12563018 TI - Regulation of signaling protein function and trafficking by the hsp90/hsp70-based chaperone machinery. AB - Nearly 100 proteins are known to be regulated by hsp90. Most of these substrates or "client proteins" are involved in signal transduction, and they are brought into complex with hsp90 by a multiprotein hsp90/hsp70-based chaperone machinery. In addition to binding substrate proteins at the chaperone site(s), hsp90 binds cofactors at other sites that are part of the heterocomplex assembly machinery as well as immunophilins that connect assembled substrate*hsp90 complexes to protein trafficking systems. In the 5 years since we last reviewed this subject, much has been learned about hsp90 structure, nucleotide-binding, and cochaperone interactions; the most important concept is that ATP hydrolysis by an intrinsic ATPase activity results in a conformational change in hsp90 that is required to induce conformational change in a substrate protein. The conformational change induced in steroid receptors is an opening of the steroid-binding cleft so that it can be accessed by steroid. We have now developed a minimal system of five purified proteins-hsp90, hsp70, Hop, hsp40, and p23- that assembles stable receptor*hsp90 heterocomplexes. An hsp90*Hop*hsp70*hsp40 complex opens the cleft in an ATP-dependent process to produce a receptor*hsp90 heterocomplex with hsp90 in its ATP-bound conformation, and p23 then interacts with the hsp90 to stabilize the complex. Stepwise assembly experiments have shown that hsp70 and hsp40 first interact with the receptor in an ATP-dependent reaction to produce a receptor*hsp70*hsp40 complex that is "primed" to be activated to the steroid binding state in a second ATP-dependent step with hsp90, Hop, and p23. Successful use of the five-protein system with other substrates indicates that it can assemble signal protein*hsp90 heterocomplexes whether the substrate is a receptor, a protein kinase, or a transcription factor. This purified system should facilitate understanding of how eukaryotic hsp70 and hsp90 work together as essential components of a process that alters the conformations of substrate proteins to states that respond in signal transduction. PMID- 12563019 TI - Renal dopamine receptors and hypertension. AB - Dopamine has been recognized as an important modulator of central as well as peripheral physiologic functions in both humans and animals. Dopamine receptors have been identified in a number of organs and tissues, which include several regions within the central nervous system, sympathetic ganglia and postganglionic nerve terminals, various vascular beds, the heart, the gastrointestinal tract, and the kidney. The peripheral dopamine receptors influence cardiovascular and renal function by decreasing afterload and vascular resistance and promoting sodium excretion. Within the kidney, dopamine receptors are present along the nephron, with highest density on proximal tubule epithelial cells. It has been reported that there is a defective dopamine receptor, especially D(1) receptor function, in the proximal tubule of various animal models of hypertension as well as in humans with essential hypertension. Recent reports have revealed the site of and the molecular mechanisms responsible for the defect in D(1) receptors in hypertension. Moreover, recent studies have also demonstrated that the disruption of various dopamine receptor subtypes and their function produces hypertension in rodents. In this review, we present evidence that dopamine and dopamine receptors play an important role in regulating renal sodium excretion and that defective renal dopamine production and/or dopamine receptor function may contribute to the development of various forms of hypertension. PMID- 12563020 TI - The effects of dietary n-3 polyunsaturated fatty acids delivered in chylomicron remnants on the transcription of genes regulating synthesis and secretion of very low-density lipoprotein by the liver: modulation by cellular oxidative state. AB - The influence of chylomicron remnants enriched in n-3 or n-6 polyunsaturated fatty acids (PUFA) (derived from fish or corn oil, respectively) on the expression of mRNA for four genes involved in the regulation of the synthesis, assembly, and secretion of very-low-density lipoprotein (VLDL) in the liver was investigated in normal rat hepatocytes and after manipulation of the cellular oxidative state by incubation with N-acetyl cysteine (NAC) or CuSO(4). The four genes investigated were those encoding apolipoprotein B (apoB), the microsomal triacylglycerol transfer protein (MTP), and the enzymes acyl coenzyme A:diacylglycerol acyltransferase (DGAT) and acyl coenzyme A:cholesterol acyltransferase 2 (ACAT2), which play a role in the regulation of triacylglycerol and cholesteryl ester synthesis, respectively. mRNA levels for apoB, MTP, and DGAT were unaffected by either fish or corn oil chylomicron remnants, but the amount of ACAT2 mRNA was significantly reduced after incubation of the hepatocytes with fish oil remnants as compared with corn oil remnants or without remnants. These findings indicate that the delivery of dietary n-3 PUFA to hepatocytes in chylomicron remnants downregulates the expression of mRNA for ACAT2, and this may play a role in their inhibition of VLDL secretion. However, when the cells were shifted into a pro-oxidizing or pro-reducing state by pretreatment with CuSO(4) (1 mM) or NAC (5 mM) for 24 hr, levels of mRNA for MTP were increased by about 2- or 4-fold, respectively, by fish oil remnants, whereas corn oil remnants had no significant effect. Fish oil remnants also caused a smaller increase in apoB mRNA in comparison with corn oil remnants in NAC-treated cells (+38%). These changes would be expected to lead to increased VLDL secretion rather than the decrease associated with dietary n-3 PUFA in normal conditions. These findings suggest that relatively minor changes in cellular redox levels can have a major influence on important liver functions such as VLDL synthesis and secretion. PMID- 12563021 TI - Maternal hyperglycemia leads to gender-dependent deficits in learning and memory in offspring. AB - Pregnancy in the diabetic woman has long been associated with an increased risk of congenital malformation in the offspring. However, little is known about the effects of maternal diabetes on development of the central nervous system. To begin to gain an understanding of this problem, diabetes was induced in adult female Sprague-Dawley rats by injection with streptozotocin. Only animals with serum glucose levels greater than 200 mg/dl were used. Diabetic and control females were bred, and all newborn pups were cross-fostered to nondiabetic mothers. At 60 days of age, pups were tested in an elevated plus-maze to assess differences in emotionality and anxiety. There were no significant differences between offspring of diabetic dams and controls on this measure. All pups were then housed individually, put on food restriction, and maintained at 85% of their ad libitum weight. They were then trained in a Lashley III maze, which assesses learning and retention capability. The female offspring of diabetic dams performed poorer than controls, a finding that was supported by inhibitory avoidance data from a separate group of animals. All animals were then trained in a radial-arm maze. Results failed to find differences between experimental and control animals. It was concluded that the diabetic intrauterine environment has gender-specific effects on central nervous system development. PMID- 12563022 TI - Pyrroloquinoline quinone improves growth and reproductive performance in mice fed chemically defined diets. AB - Growth, reproductive performance, and indices of collagen maturation and expression were investigated in Balb/c mice fed chemically defined, amino acid based diets with or without the addition 6 micro Mpyrroloquinoline quinone (PQQ)/kg diet. The diets were fed to virgin mice for 8 weeks before breeding. At weaning, the pups from successful pregnancies were fed the same diet as their respective dams. Reproductive performance was compromised in mice fed diets devoid of PQQ, and their offspring grew at slower rates than offspring from mice fed diets supplemented with PQQ. Successful mating (confirmed vaginal plugs) was not affected by the presence or absence of PQQ; however, pup viability (number of pups at parturition/number of pups at Day 4 of lactation) was decreased in PQQ deprived mice. Conception (percentage of females giving live births) and fertility (percentage of births) were also decreased in PQQ-deprived mice. The slower rates of growth in offspring from PQQ-deprived mice were associated with decreased steady-state mRNA levels for Type I procollagen alpha(1)-chains in skin and lungs from neonatal mice. Values for lysyl oxidase accumulation as protein in PQQ-deficient mice also tended to be lower than corresponding values from PQQ supplemented or -replete mice. Skin collagen solubility was increased in PQQ deprived mice. These results indicate that PQQ supplementation can improve reproductive performance, growth, and may modulate indices of neonatal extracellular matrix production and maturation in mice fed chemically defined, but otherwise nutritionally complete diets. PMID- 12563023 TI - A measure of heart rate variability is sensitive to orthostatic challenge in women with chronic fatigue syndrome. AB - The use of symptoms generated by head up tilt (HUT) is not a useful tool in identifying chronic fatigue syndrome (CFS). We investigated whether heart rate variability (HRV) assessed early during HUT might be useful. A sample of 46 female subjects (24 with CFS and 22 sedentary, age-matched healthy controls; CON) who had exhibited no difference in time to syncope during tilt was examined for HRV responses to 10 min of 70 degrees HUT after 5 min of baseline in the supine position. HRV data were analyzed by the method of coarse graining spectral analysis. Variables compared between groups included mean and standard deviation (SD(RRI)) of RR intervals (RRI), amplitudes of low- (A(LF); 0.04-0.15 Hz) and high-frequency (A(HF); >0.15 Hz) harmonic as well as aperiodic, fractal (A(FR); 1/f(beta)) spectral components, the spectral exponent beta, and the difference in these values between baseline and HUT for each subject. In the supine baseline, only mean RRI was significantly (P < 0.01) lower in CFS than in CON. During HUT, however, mean RRI (P < 0.01), SD(RRI) (P < 0.01), A(HF) (P < 0.05), and A(FR) (P < 0.01) were significantly lower in CFS than in CON. When the difference in values between baseline and HUT for each subject was examined, only the difference for A(FR) (deltaA(FR)) was significantly (P < 0.01) lower in CFS than in CON, suggesting that A(FR)is a disease-specific response of HRV to HUT. When a cut-off level was set to deltaA(FR) = -2.7 msec, the sensitivity and the specificity in differentiating CFS from controls were 90% and 72%, respectively. The data suggest that a decrease in aperiodic fractal component of HRV in response to HUT can be used to differentiate patients with CFS from CON. PMID- 12563024 TI - Leptin constrains phospholipase C-protein kinase C-induced insulin secretion via a phosphatidylinositol 3-kinase-dependent pathway. AB - Leptin-deficient Lep(ob)/Lep(ob)mice hypersecrete insulin in response to acetylcholine stimulation of the phospholipase C-protein kinase C (PLC-PKC) pathway, and leptin constrains this hypersecretion. Leptin has been reported to activate phosphatidylinositol 3-kinase (PI 3-K) and subsequently phosphodiesterase (PDE) to impair protein kinase A (PKA)-induced insulin secretion from cultured islets of neonatal rats. We determined if PKA-induced insulin secretion was also hyperresponsive in islets from Lep(ob)/Lep(ob)mice, and if leptin impaired this pathway in islets from these mice. Additionally, the possible role for PI 3-K and PDE in leptin-induced control of acetylcholine induced insulin secretion was examined. Stimulation of insulin secretion with GLP 1, forskolin (an activator of adenylyl cyclase), or IBMX (an inhibitor of PDE) did not cause hypersecretion of insulin from islets of young Lep(ob)/Lep(ob)mice, and leptin did not inhibit GLP-1-induced insulin secretion from islets of these mice. Inhibition of PDE with IBMX also did not block leptin-induced inhibition of acetylcholine-mediated insulin secretion from islets of Lep(ob)/Lep(ob)mice. But, preincubation of islets with wortmannin, an inhibitor of PI 3-K activity, blocked the ability of leptin to constrain acetylcholine-induced insulin secretion from islets of Lep(ob)/Lep(ob)mice. We conclude that the capacity of the PKA pathway to stimulate insulin secretion is not increased in islets from young Lep(ob)/Lep(ob)mice, and that leptin does not regulate this pathway in islets from mice. Leptin may stimulate PI 3-K to constrain PLC-PKC-induced insulin secretion from islets of Lep(ob)/Lep(ob)mice. PMID- 12563025 TI - Leptin administration normalizes insulin secretion from islets of Lep(ob)/Lep(ob) mice by food intake-dependent and -independent mechanisms. AB - Leptin-deficient Lep(ob)/Lep(ob) mice exhibit elevations in plasma insulin early in development. The present study tested the hypothesis that absence of leptin during neonatal development permanently programs islets from these mice to hypersecrete insulin. Administration of leptin for 8 days to young adult Lep(ob)/Lep(ob) mice normalized their food intake, plasma insulin concentration, and insulin secretion in response to glucose, acetylcholine, and leptin. Restriction of food intake per se of Lep(ob)/Lep(ob) mice lowered, but did not normalize, plasma insulin concentrations. Food-restricted Lep(ob)/Lep(ob) mice continued to hypersecrete insulin in response to glucose, but islets from these mice did not hyperrespond to acetylcholine or respond to leptin as occurs in ad libitum-fed Lep(ob)/Lep(ob) mice. We conclude that neonatal leptin deficiency does not permanently program islets from mice to hypersecrete insulin. The hyperphagia associated with leptin deficiency contributes substantially to the hypersecretion of insulin, but leptin also appears to have more direct effects on regulation of insulin secretion. PMID- 12563026 TI - Phytosterols affect endocrinology and metabolism of the field vole (Microtus agrestis). AB - Phytosterols or plant sterols (PS) enter the ecosystem via pulp mill effluents. They are also consumed by the general population of developed countries in natural remedies and margarines to lower elevated serum cholesterol levels. This study screened the endocrine and enzymatic parameters of the field vole (Microtus agrestis) for the effects of subchronic PS exposure at three doses (0, 5, or 50 mg of PS kg(-1) day(-1)). PS at 5 or 50 mg kg(-1) day(-1) decreased the relative liver weight of the voles. The kidney glycogen phosphorylase activity decreased at 5 or 50 mg kg(-1) day(-1), but the liver glycogen phosphorylase activity increased at 5 mg kg(-1) day(-1). The plasma estradiol and testosterone concentrations of males were higher due to PS supplement at 5 mg kg(-1) day(-1). This can be due to increased sex steroid synthesis from PS precursors. Biotransformation enzyme activities were not affected. PS caused multiple, previously unreported effects that were more pronounced at a low dose. As 5 mg PS kg(-1) day(-1) is the recommended dose for various health products, a thorough risk assessment of the effects and interactions of PS is warranted. PMID- 12563027 TI - Cardiac norepinephrine release: modulation by ovariectomy and estrogen. AB - Previously, we have demonstrated that in contrast to male rats, female rats do not show an age-related reduction of depolarization-elicited norepinephrine (NE) release from cardiac synaptosomes (resealed nerve terminals). These results suggest that sex hormones such as estrogen may modulate NE release from cardiac synaptosomes prepared from female rats. The present study was designed to test the hypotheses that long-term estrogen depletion, resulting from ovariectomy, and estrogen replacement alters depolarization-elicited NE release from cardiac synaptosomes. Female F344 rats were divided into two groups, one of which underwent bilateral ovariectomy, whereas the other underwent a sham operation. Three ovariectomized subgroups received daily injections of conjugated equine estrogens, delta8,9-dehydroestrone or 17 alpha-dihydroequilenin. Another ovariectomized control subgroup and the sham-operated animals received daily injections of vehicle. After 90 or 270 days of treatment, the animals were sacrificed. Cardiac synaptosomes were prepared from each heart, incubated with [(3)H]-NE, and used to evaluate NE release capacity by exposure to 50 mM K(+). The effectiveness of the ovariectomy and the estrogenic actions of the test compounds was confirmed by evaluating vaginal smears, determining uterine weights, and measuring serum luteinizing hormone (LH) concentrations. Ovariectomy (after both 90 and 270 days) significantly increased depolarization-induced NE release compared with sham-operated rats. Treatment with all three estrogenic preparations reduced NE release in ovariectomized rats to values similar to those observed in sham-operated animals. Interestingly, NE release rates from rats treated with conjugated estrogens for 270 but not 90 days were significantly below that observed in age-matched sham animals. These results demonstrate that estrogen modulates depolarization-elicited NE release from cardiac nerve terminals. Such modulation may represent a protective action by estrogen at the cardiac synapse. PMID- 12563028 TI - Stretch-activated atrial natriuretic peptide secretion in atria with heat shock protein 70 overexpression. AB - The mechanical overload of the heart is known to induce the expression of atrial natriuretic peptide (ANP) and heat-shock protein 70 (HSP70) in the heart. However, the relationship between these two substances remains unknown. In the present study, we characterized ANP secretion from mouse atria and examined a possible role that HSP70 may play in the regulation of ANP synthesis and secretion by using atria in transgenic mice in which HSP70 was overexpressed. We generated transgenic mice harboring the human HSP70gene under the transcriptional control of human myosin heavy-chain promoter. In these mice, the transgene was overexpressed in the heart. Both atrial ANP messenger RNA and its concentration in the HSP70 transgenic mice were measured; these were not significantly different from those in wild-type mice. In isolated perfused nonbeating atria, basal secretion of ANP was similar in both groups. When atrial volume was increased by changing atrial pressure, extracellular fluid (ECF) translocation and ANP secretion proportionately increased. Changes in atrial volume and ECF translocation and ANP secretion were positively correlated. However, these parameters did not significantly differ between the two groups. Endothelin-1 (ET 1), the strongest paracrine stimulus of ANP secretion, accentuated stretch activated ANP secretion without significantly changing mechanically stimulated ECF translocation, as compared with that in the wild-type mice. The increased ANP secretion due to ET-1 in the transgenic mice was similar to that in the wild-type mice. The results suggest that both atrial stretching and ET-1 are important stimuli to ANP secretion from mouse atria, and the responsiveness of the ANP system to those stimuli are unlikely coupled to the pathway involving HSP70. PMID- 12563029 TI - Enlargement of interscapular brown adipose tissue in growth hormone antagonist transgenic and in growth hormone receptor gene-disrupted dwarf mice. AB - Growth hormone (GH) acts on adipose tissue by accelerating fat expenditure, preventing triglyceride accumulation, and facilitating lipid mobilization. To investigate whether GH is involved in the development and metabolism of interscapular brown adipose tissue (BAT), a site of nonshivering thermogenesis, we employed three lines of transgenic mice. Two of the lines are dwarf due to expression of a GH antagonist (GHA) or disruption of the GH receptor/binding protein gene. A third mouse line is giant due to overexpression of a bovine GH (bGH) transgene. We have found that the body weights of those animals are proportional to their body lengths at 10 weeks of age. However, GHA dwarf mice tend to catch up with the nontransgenic (NT) littermates in body weight but not in body length at 52 weeks of age. The increase of body mass index (BMI) for GHA mice accelerates rapidly relative to controls as a function of age. We have also observed that BAT in both dwarf mouse lines but not in giant mice is enlarged in contrast to nontransgenic littermates. This enlargement occurs as a function of age. Northern analysis suggests that BAT can be a GH-responsive tissue because GHR/BP mRNAs were found there. Finally, the level of uncoupling protein-1 (UCP1) RNA was found to be higher in dwarf mice and lower in giant animals relative to controls, suggesting that GH-mediated signaling may negatively regulate UCP1 gene expression in BAT. PMID- 12563030 TI - Kep1 interacts genetically with dredd/caspase-8, and kep1 mutants alter the balance of dredd isoforms. AB - The Drosophila kep1 gene encodes an RNA binding protein related to the murine QUAKING apoptotic inducer. We have previously shown that kep1 can induce apoptosis when transfected into different cell lines. To better define the role of Kep1 in apoptosis, we generated kep1 null flies. These flies were viable, but females displayed reduced fertility, with approximately half of the eggs laid from kep1- homozygotes failing to hatch. In addition, loss of kep1 suppressed GMR rpr-mediated apoptosis in the Drosophila eye, and kep1 mutant flies had increased susceptibility to Escherichia coli infection. We found that Kep1 bound dredd RNA in vitro, and that extracts prepared from kep1 mutant ovaries had markedly reduced proteolytic cleavage activity toward the caspase-8 target substrate IETD 7-amino-4-trifluoromethyl coumarin. We observed increased levels of the beta isoform of dredd mRNA in kep1 mutants as compared with wild-type. Taken together, our results suggest that Kep1 regulates apoptosis by influencing the processing of dredd RNA. PMID- 12563031 TI - Facultative bacterial symbionts in aphids confer resistance to parasitic wasps. AB - Symbiotic relationships between animals and microorganisms are common in nature, yet the factors controlling the abundance and distributions of symbionts are mostly unknown. Aphids have an obligate association with the bacterium Buchnera aphidicola (the primary symbiont) that has been shown to contribute directly to aphid fitness. In addition, aphids sometimes harbor other vertically transmitted bacteria (secondary symbionts), for which few benefits of infection have been previously documented. We carried out experiments to determine the consequences of these facultative symbioses in Acyrthosiphon pisum (the pea aphid) for vulnerability of the aphid host to a hymenopteran parasitoid, Aphidius ervi, a major natural enemy in field populations. Our results show that, in a controlled genetic background, infection confers resistance to parasitoid attack by causing high mortality of developing parasitoid larvae. Compared with uninfected controls, experimentally infected aphids were as likely to be attacked by ovipositing parasitoids but less likely to support parasitoid development. This strong interaction between a symbiotic bacterium and a host natural enemy provides a mechanism for the persistence and spread of symbiotic bacteria. PMID- 12563032 TI - Crystal structure of a photoactive yellow protein from a sensor histidine kinase: conformational variability and signal transduction. AB - Photoactive yellow protein (E-PYP) is a blue light photoreceptor, implicated in a negative phototactic response in Ectothiorhodospira halophila, that also serves as a model for the Per-Arnt-Sim superfamily of signaling molecules. Because no biological signaling partner for E-PYP has been identified, it has not been possible to correlate any of its photocycle intermediates with a relevant signaling state. However, the PYP domain (Ppr-PYP) from the sensor histidine kinase Ppr in Rhodospirillum centenum, which regulates the catalytic activity of Ppr by blue light absorption, may allow such issues to be addressed. Here we report the crystal structure of Ppr-PYP at 2 A resolution. This domain has the same absorption spectrum and similar photocycle kinetics as full length Ppr, but a blue-shifted absorbance and considerably slower photocycle than E-PYP. Although the overall fold of Ppr-PYP resembles that of E-PYP, a novel conformation of the beta 4-beta 5 loop results in inaccessibility of Met-100, thought to catalyze chromophore reisomerization, to the chromophore. This conformation also exposes a highly conserved molecular surface that could interact with downstream signaling partners. Other structural differences in the alpha 3-alpha 4 and beta 4-beta 5 loops are consistent with these regions playing significant roles in the control of photocycle dynamics and, by comparison to other sensory Per-Arnt-Sim domains, in signal transduction. Because of its direct linkage to a measurable biological output, Ppr-PYP serves as an excellent system for understanding how changes in photocycle dynamics affect signaling by PYPs. PMID- 12563033 TI - PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. AB - Streptomycetes are high G+C Gram-positive, antibiotic-producing, mycelial soil bacteria. The 8.7-Mb Streptomyces coelicolor genome was previously sequenced by using an ordered library of Supercos-1 clones. Here, we describe an efficient procedure for creating precise gene replacements in the cosmid clones by using PCR targeting and lambda-Red-mediated recombination. The cloned Streptomyces genes are replaced with a cassette containing a selectable antibiotic resistance and oriT(RK2) for efficient transfer to Streptomyces by RP4-mediated intergeneric conjugation. Supercos-1 does not replicate in Streptomyces, but the clones readily undergo double-crossover recombination, thus creating gene replacements. The antibiotic resistance cassettes are flanked by yeast FLP recombinase target sequences for removal of the antibiotic resistance and oriT(RK2) to generate unmarked, nonpolar mutations. The technique has been used successfully by >20 researchers to mutate around 100 Streptomyces genes. As an example, we describe its application to the discovery of a gene involved in the production of geosmin, the ubiquitous odor of soil. The gene, Sco6073 (cyc2), codes for a protein with two sesquiterpene synthase domains, only one of which is required for geosmin biosynthesis, probably via a germacra-1 (10) E,5E-dien-11-ol intermediate generated by the sesquiterpene synthase from farnesyl pyrophosphate. PMID- 12563034 TI - Histidine-rich amphipathic peptide antibiotics promote efficient delivery of DNA into mammalian cells. AB - Gene delivery has shown potential in a wide variety of applications, including basic research, therapies for genetic and acquired diseases, and vaccination. Most available nonviral systems have serious drawbacks such as the inability to control and scale the production process in a reproducible manner. Here, we demonstrate a biotechnologically feasible approach for gene delivery, using synthetic cationic amphipathic peptides containing a variable number of histidine residues. Gene transfer to different cell lines in vitro was achieved with an efficiency comparable to commercially available reagents. We provide evidence that the transfection efficiency depends on the number and positioning of histidine residues in the peptide as well as on the pH at which the in-plane to transmembrane transition takes place. Endosomal acidification is also required. Interestingly, even when complexed to DNA these peptides maintain a high level of antibacterial activity, opening the possibility of treating the genetic defect and the bacterial infections associated with cystic fibrosis with a single compound. Thus, this family of peptides represents a new class of agents that may have broad utility for gene transfer and gene therapy applications. PMID- 12563035 TI - JNK-interacting protein-1 promotes transcription of A beta protein precursor but not A beta precursor-like proteins, mechanistically different than Fe65. AB - Processing of the amyloid beta protein precursor (A beta PP) by the beta and gamma secretases leads to the production of two small peptides, amyloid beta and the A beta PP intracellular domain (AID, or called elsewhere AICD). Whereas the role of amyloid beta in the pathogenesis of Alzheimer's disease has been studied extensively, only recently has information begun to accumulate as to the role of AID. Functions identified for AID include its ability to trigger apoptosis and a role in regulating gene transcription, particularly in combination with the A beta PP binding protein Fe65. Here, we report that AID in combination with Janus kinase interacting protein-1 (JIP-1) can activate gene expression. We demonstrate that the mechanism is different from activation in combination with Fe65 by first showing that although Fe65 enters the nucleus in the absence of full-length A beta PP, JIP-1 does not. Additionally, JIP-1-induced activation is Tip60 independent, whereas a complex with AID, Fe65, and Tip60 is formed for Fe65 induced activation. Finally, and probably most interestingly, we show that although the A beta PP family members APLP1 and APLP2 (for amyloid beta precursor like protein) can cause activation in combination with Fe65, APLP1 and APLP2 show little or no activation in combination with JIP-1. This activity for the AID fragment may help explain the unique functions of A beta PP relative to its other family members, and changes in gene expression found in Alzheimer's disease. PMID- 12563036 TI - Decreased embryonic retinoic acid synthesis results in a DiGeorge syndrome phenotype in newborn mice. AB - Retinoic acid (RA), the active derivative of vitamin A, is involved in various developmental and homeostatic processes. To define whether certain developmental events are particularly sensitive to a decrease in embryonic RA levels, we generated mice bearing a hypomorphic allele of the RA-synthesizing enzyme Raldh2. The resulting mutant mice, which die perinatally, exhibit the features of the human DiGeorge syndrome (DGS) with heart outflow tract septation defects and anomalies of the aortic arch-derived head and neck arteries, laryngeal-tracheal cartilage defects, and thymus/parathyroid aplasia or hypoplasia. Analysis of Raldh2 hypomorph embryos reveal selective defects of the posterior (third to sixth) branchial arches, including absence or hypoplasia of the corresponding aortic arches and pharyngeal pouches, and local down-regulation of RA-target genes. Thus, a decreased level of embryonic RA (through genetic and/or nutritional causes) could represent a major modifier of the expressivity of human 22q11del-associated DiGeorge/velocardiofacial syndromes and, if severe enough, could on its own lead to the clinical features of the DiGeorge syndrome. PMID- 12563037 TI - The strategy for coupling the RanGTP gradient to nuclear protein export. AB - The Ran GTPase plays critical roles in both providing energy for and determining the directionality of nucleocytoplasmic transport. The mechanism that couples the RanGTP gradient to nuclear protein export will determine the rate of and limits to accumulation of export cargoes in the cytoplasm, but is presently unknown. We reasoned that plausible coupling mechanisms could be distinguished by comparing the rates of reverse motion of export cargoes through the nuclear pore complex (NPC) with the predictions of a mathematical model. Measurement of reverse export rates in Xenopus oocytes revealed that nuclear export signals can facilitate RanGTP-dependent cargo movement into the nucleus against the RanGTP gradient at rates comparable to export rates. Although export cargoes with high affinity for their receptor are exported faster than those with low affinity, their reverse transport is also greater. The ratio of the rates of reverse and forward export of a cargo is proportional to its rate of diffusion through the NPC, i.e., to the ability of the cargo to penetrate the NPC permeability barrier. The data substantiate a diffusional mechanism of coupling and suggest the existence of a high concentration of RanGTP-receptor complexes within the NPC that decreases sharply at the cytoplasmic boundary of the NPC permeability barrier. PMID- 12563038 TI - Movement disorders in children. PMID- 12563039 TI - Neutrophil abnormalities. PMID- 12563040 TI - Index of suspicion. PMID- 12563041 TI - Managing temper tantrums. PMID- 12563042 TI - Question from the clinician: fluoridated water. PMID- 12563043 TI - Prevention of hospital-acquired hyponatremia: a case for using isotonic saline. AB - OBJECTIVE: The current standard of care in pediatrics is to administer hypotonic saline in maintenance parenteral fluids. The safety of this approach has never been evaluated. METHODS: A review of the literature reveals that the administration of hypotonic fluids is potentially dangerous and may not be physiologic for the hospitalized child. RESULTS: There have been >50 reported cases of neurologic morbidity and mortality, including 26 deaths, in the past 10 years resulting from hospital-acquired hyponatremia in children who were receiving hypotonic parenteral fluids. Common childhood conditions requiring parenteral fluids, such as pulmonary and central nervous system infections, dehydration, and the postoperative state, are associated with a nonosmotic stimulus for antidiuretic hormone production, which can lead to free water retention and hyponatremia. Children are at particularly high risk of developing symptomatic hyponatremia as they have a larger brain-to-skull size ratio. CONCLUSIONS: The administration of isotonic saline in maintenance parenteral fluids is the most important prophylactic measure that can be taken to prevent the development of hyponatremia in children who receive parenteral fluids. PMID- 12563044 TI - Misconceptions about colds and predictors of health service utilization. AB - OBJECTIVE: Colds accounted for 1.6 million emergency department (ED) visits and 25 million ambulatory visits by children and adults in 1998. Although most colds are caused by viruses and do not require medical intervention, many families seek health care for the treatment of colds. Parental misconceptions about the cause and appropriate treatment of colds may contribute to unnecessary health service utilization. The objective of this study was to determine predictors of reported ED use and ambulatory care use for colds among families with young children. METHODS: This study was an observational, prospective cohort study to determine attack rates for respiratory illnesses within families that have at least 1 child who is 6 months to 5 years of age and enrolled in out-of-home child care. Families were randomly selected from 5 pediatric practices in Massachusetts and were considered eligible when the child was enrolled in child care with at least 5 other children for >or=10 hours per week. Enrolled families were asked to complete a survey that assessed knowledge about colds, antibiotic indications, and frequency of health service utilization. Predictors of self-reported use of health care services were assessed in multivariate logistic regression models. RESULTS: Of the 261 families enrolled in the study, 197 families (75%) returned completed surveys. Although 93% of parents understood that viruses caused colds, 66% of parents also believed that colds were caused by bacteria. Fifty-three percent believed that antibiotics were needed to treat colds. Parents reported that they would visit the ED (23%) or their doctor's office (60%) when their child had a cold. Predictors of ED use on multivariate analysis included Medicaid insurance (odds ratio [OR]: 17.6 [2.2-139.3]), history of wheezing (OR: 18.3 [4.4 75.8]), and belief that antibiotics treat colds (OR: 4.2 [1.4-12.9]). Predictors of ambulatory care use included parent younger than 30 years (OR: 10.0 [1.6 64.3]), history of wheezing (OR: 5.6 [1.1-29.7]), and belief that antibiotics treat colds (OR: 3.8 [1.7-8.5]). CONCLUSIONS: Misconceptions about the appropriate treatment of colds are predictive of increased health service utilization. Targeted educational interventions for families may reduce inappropriate antibiotic-seeking behavior and unnecessary health service utilization for colds. PMID- 12563045 TI - Geographic variation in the prevalence of stimulant medication use among children 5 to 14 years old: results from a commercially insured US sample. AB - OBJECTIVE: The purpose of this study was to evaluate geographic variation in the prevalence of prescription stimulant use and predictors of use among a nationally representative, commercially insured population 5 to 14 years old. METHODS: Prescription claims activity from January 1, 1999 through December 31, 1999 for a continuously eligible population 5 to 14 years old was evaluated. Age-gender adjusted prevalence rates were estimated for each state. Multivariate logistic regression using hierarchical linear modeling was used to evaluate the impact of age, gender, number of child dependents, and region of the country on stimulant prevalence. The contextual effects of urban or rural residence, median income, percent white, and physician rate per 100 000 residents were also controlled for. RESULTS: The 1-year prevalence of stimulant treatment for the entire study sample was 4.2%. Multivariate logistic regression indicated that stimulant prescription use was positively associated with age, male gender, fewer child dependents, living in higher income communities, and living in communities with greater percent white. Compared with children living in the Western region of the country, children living in the Midwest and South were 1.55 (99% confidence interval: 1.28-1.87) and 1.71 (99% confidence interval: 1.42-2.06) times more likely to consume at least 1 stimulant medication, respectively. Differences in stimulant prevalence across urban and rural residence were also noted. CONCLUSIONS: Geographic variation in the prevalence of stimulant use exists nationally, despite controlling for important predictors of use including age and gender. Possible reasons for the variation are discussed as are calls for additional research. PMID- 12563046 TI - Treatment of term infants with head cooling and mild systemic hypothermia (35.0 degrees C and 34.5 degrees C) after perinatal asphyxia. AB - OBJECTIVE: To assess the safety of selective head cooling in birth-asphyxiated term newborn infants while maintaining the rectal temperature at 35.0 degrees C or 34.5 degrees C. METHODS: Twenty-six term infants with Apgar 2499 g) subjects in late adolescence, and to determine whether growth restriction in utero was related to BP in VLBW survivors at this age. METHODS: This was a cohort study of 210 preterm survivors with birth weights <1501 g born from January 1, 1977, to March 31, 1982, and 60 randomly selected NBW subjects from the Royal Women's Hospital, Melbourne. BP was measured at 18+ years of age in 156 (74%) VLBW subjects and 38 (63%) NBW subjects with both a standard mercury sphygmomanometer and an ambulatory BP monitor. RESULTS: VLBW subjects had higher sphygmomanometer systolic and diastolic BPs than NBW subjects (mm Hg; mean difference [95% confidence interval]; systolic, 8.6 [3.4, 13.9]; diastolic, 4.3 [1.0, 7.6]). VLBW subjects also had significantly higher mean systolic ambulatory BPs (mm Hg; mean difference [95% confidence interval]) for the 24-hour period (4.7 [1.4, 8.0]), and for both the awake (5.0 [1.6, 8.5]) and asleep (3.6 [0.04, 7.1]) periods. There were no significant differences between the birth weight groups for any ambulatory diastolic BPs. Within the VLBW subjects, there was no significant relationship between birth weight standard deviation score and any measure of BP. CONCLUSIONS: BP was significantly higher in late adolescence in VLBW survivors than in NBW subjects. Growth restriction in utero was not significantly related to BP in VLBW survivors. PMID- 12563048 TI - Mevalonate kinase deficiency: enlarging the clinical and biochemical spectrum. AB - OBJECTIVE: Mevalonic aciduria as a result of mevalonate kinase deficiency is an inborn error of cholesterol biosynthesis characterized by dysmorphology, psychomotor retardation, progressive cerebellar ataxia, and recurrent febrile crises, usually manifesting in early infancy, accompanied by hepatosplenomegaly, lymphadenopathy, arthralgia, and skin rash. The febrile crises are similar to those observed in hyperimmunoglobulinemia D and periodic fever syndrome (HIDS). Pathogenic mutations in the mevalonate kinase gene in both disorders have demonstrated a common genetic basis. Our aim was to describe the clinical picture of adolescent patients with mevalonate kinase deficiency and to expand the clinical and biochemical spectrum of mevalonate kinase deficiency, particularly with regard to HIDS. METHODS: We report the clinical history and biochemical findings of 3 patients with mevalonic aciduria. RESULTS: In 2 siblings with mevalonic aciduria, a 15-year-old girl and a 14-year-old boy, the phenotype shifted with age. Ataxia has become the predominant clinical manifestation, whereas the febrile attacks occur less frequently but as yet have not disappeared. Both of them show marked elevations of immunoglobulin D (IgD). Psychomotor development is retarded but not regressive. Short stature developed in both patients. Additional findings include the development of retinal dystrophy and cataracts in both of them. The third patient is a 6-year-old boy who presented at the age of 5 years with cerebellar ataxia and retinal dystrophy. He is different from all known patients with mevalonic aciduria because of the mild neurologic involvement and because he has never developed febrile crises. In addition, levels of IgD were repeatedly normal. CONCLUSION: The clinical and biochemical spectrum of patients with mevalonic aciduria is heterogeneous. Manifestations of the disease seem to be age dependent, as evident from this first report of adolescent patients. In patients who survive infancy, short stature, ataxia caused by cerebellar atrophy, and ocular involvement with retinal dystrophy become predominant findings. Recurrent febrile crises seem to diminish with increasing age and may not even be an obligatory finding. Elevation of IgD is most likely a secondary phenomenon that seems to be linked to recurrent febrile crises. PMID- 12563049 TI - Childhood behavior disorders and injuries among children and youth: a population based study. AB - CONTEXT: While an association between pediatric behavioral disorders and injuries is generally acknowledged, no studies have measured the risk for injury in the context of a large, population-based study that is free of cohort biases. OBJECTIVES: To examine the association between childhood behavior disorders ([CBDs] as indicated by prescription for methylphenidate [MPH]) and a variety of injury outcomes, and to evaluate the risk for injury among these children after controlling for known demographic correlates. DESIGN: Population-based database analysis of all children in British Columbia (BC) under the age of 19 as of December 31, 1996; comparison of those who had been prescribed MPH and therefore placed in the CBD group (n = 16, 806) and those who were not (n = 1,010,067). Demographic information collected was as follows: age, sex, measures of socioeconomic status, and region of residence. OUTCOME MEASURES: Common types of childhood injury in BC: International Classification of Diseases, Ninth Revision N-codes (fractures, open wounds, poisoning/toxic effect, concussion, intracranial, burns) and E-codes (falls, postoperative complications, motor vehicle accidents, struck by object, adverse effects of drugs, suffocation, drowning). DATA SOURCE: BC Linked Health Data Set and the BC Triplicate Prescription Program. RESULTS: After controlling for known demographic correlates, odds for injury was greater among those treated with MPH and presumed to have a behavioral disorder, when injury was characterized either by type (1.67; 99% confidence interval: 1.54-1.81) or cause (1.52; 99% confidence interval: 1.40-1.66) of injury. This increased risk extended to unexpected categories of injury such as postoperative complications and adverse effects of drugs. CONCLUSIONS: Children with CBDs have >1.5 times the odds of sustaining injuries of a variety of types from a variety of causes, even after controlling for known demographic correlates, than those without behavioral disorders. The risks for these children extend beyond those that might be directly associated with impulsivity and overactivity. Injury prevention strategies aimed at this group of children and youth would be beneficial. Policy-makers should account for increased risk of a wide variety of injuries in this group of children and youth. PMID- 12563050 TI - Stunting of growth as a major feature of anorexia nervosa in male adolescents. AB - OBJECTIVE: To assess growth retardation in male adolescent patients who have a diagnosis of anorexia nervosa (AN) and the effect of weight restoration on catch up growth. METHODS: Medical charts of all male adolescent AN patients (n = 12) who were admitted to the Pediatric Psychosomatic Department at the Sheba Medical Center from January 1, 1994, to December 31, 1998, were reviewed. Height and weight measurements were obtained before the onset of AN, at admission, and thereafter routinely during hospitalization and follow-up. RESULTS: Eleven patients exhibited growth retardation during the course of their illness, as evident in a decrease in their height standard deviation score (SDS). The mean height SDS at the time of admission (-0.81 +/- 0.93) was significantly lower than the premorbid SDS (-0.21 +/- 0.91). Weight restoration resulted in accelerated linear growth (up to 2 cm/mo) in all patients. Positive weight gain (weight gain rate >1 kg/y) was associated with a mean height gain of 6.97 +/- 6.48 cm/y, whereas weight loss or failure to gain weight (weight gain rate 15% at the beginning of 1994. CONCLUSIONS: Poliovirus extraimmunization rates have fallen dramatically in association with the change-over to the all inactivated polio virus schedule. PMID- 12563055 TI - Sleep duration from infancy to adolescence: reference values and generational trends. AB - OBJECTIVE: The main purpose of the present study was to calculate percentile curves for total sleep duration per 24 hours, for nighttime and for daytime sleep duration from early infancy to late adolescence to illustrate the developmental course and age-specific variability of these variables among subjects. METHODS: A total of 493 subjects from the Zurich Longitudinal Studies were followed using structured sleep-related questionnaires at 1, 3, 6, 9, 12, 18, and 24 months after birth and then at annual intervals until 16 years of age. Gaussian percentiles for ages 3 months to 16 years were calculated for total sleep duration (time in bed) and nighttime and daytime sleep duration. The mean sleep duration for ages 1 to 16 years was estimated by generalized additive models based on the loess smoother; a cohort effect also had to be included. The standard deviation (SD) was estimated from the loess smoothed absolute residuals from the mean curve. For ages 3, 6, and 9 months, an alternative approach with a simple model linear in age was used. For age 1 month, empirical percentiles were calculated. RESULTS: Total sleep duration decreased from an average of 14.2 hours (SD: 1.9 hours) at 6 months of age to an average of 8.1 hours (SD: 0.8 hours) at 16 years of age. The variance showed the same declining trend: the interquartile range at 6 months after birth was 2.5 hours, whereas at 16 years of age, it was only 1.0 hours. Total sleep duration decreased across the studied cohorts (1974 1993) because of increasingly later bedtime but unchanged wake time across decades. Consolidation of nocturnal sleep occurred during the first 12 months after birth with a decreasing trend of daytime sleep. This resulted in a small increase of nighttime sleep duration by 1 year of age (mean 11.0 +/- 1.1 hours at 1 month to 11.7 +/- 1.0 hours at 1 year of age). The most prominent decline in napping habits occurred between 1.5 years of age (96.4% of all children) and 4 years of age (35.4%). CONCLUSIONS: Percentile curves provide valuable information on developmental course and age-specific variability of sleep duration for the health care professional who deals with sleep problems in pediatric practice. PMID- 12563056 TI - Are hospital characteristics associated with parental views of pediatric inpatient care quality? AB - OBJECTIVE: Patient assessments of care are increasingly being considered an important dimension of quality of care. Few studies have examined the types and extent of problems identified by parents in the care of hospitalized children and whether hospital characteristics are associated with some of these problems. The objective of this study was to describe the quality of pediatric inpatient care as perceived by parents of hospitalized children and test whether hospital characteristics (academic status, market competition, freestanding children's hospital) are associated with variations in quality. METHODS: We performed a cross-sectional analysis of surveys from 6030 parents of children who were discharged for a medical condition from 38 hospitals that used the Picker Institute's Pediatric Inpatient Survey. The Pediatric Inpatient Survey measures 7 dimensions of inpatient care quality: partnership, coordination, information to parent, information to child, physical comfort, confidence and trust, and continuity and transition. Our main outcome measures included an overall quality of care rating (1 = poor, 5 = excellent), as well as overall and dimension specific problem scores (0 = no problems, 100 = problems with 100% of processes asked about in the survey). We used Pearson correlation to determine the strength of association between the overall quality of care rating and dimension problem scores. We tested for associations between hospital characteristics and problem scores using linear regression models, controlling for patient health status and other socioeconomic status variables. RESULTS: Parents on average rated their child's care as very good (mean: 4.2) but reported problems with 27% of the survey's hospital process measures. Information to the child (33%) and coordination of care (30%) had the highest problem rates. Parent communication problems correlated most strongly with overall quality of care ratings (r = 0.49). Parents of children who were hospitalized at academic health centers (AHCs) reported 4% more problems overall (29.8% vs 25.5%) and almost 9% more problems with coordination of care (34.1% vs 25.6%) compared with those at non AHCs. Parents in more competitive markets reported almost 3% more problems than those in the less competitive ones (28.9% vs 26.3%). The freestanding children's hospital classification was not associated with overall problem scores. We found wide variation in problem scores by hospital, even among AHCs. Hospital and patient characteristics explained only 6% of the variance in problem scores. CONCLUSIONS: Despite high subjective ratings of quality of care, measures of specific processes of care reveal significant variations among hospitals and identify areas with opportunities for improvement. Improving the quality of communication with the parent of a hospitalized child may have the most positive impact on a hospital's overall quality of care rating. AHCs and hospitals in more competitive markets may be more prone to problems. With wide variation in parental perceptions of hospital quality of care, a systems analysis of individual hospitals may provide strategies for hospitals to deliver higher quality care. PMID- 12563057 TI - Parental factors, mass media influences, and the onset of eating disorders in a prospective population-based cohort. AB - OBJECTIVE: To identify risk factors for eating disorders. METHODS: A community cohort study was conducted in Navarra, Spain. A region-wide representative sample of 2862 girls who were 12 to 21 years of age completed the Eating Attitudes Test (40-item version) and other questionnaires in 1997. Girls who scored high in the Eating Attitudes Test-40 were interviewed by a psychiatrist who applied Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria to diagnose prevalent cases of eating disorders. Girls who were free of any eating disorder in 1997 were reassessed after 18 months of follow-up using the same methods. RESULTS: Ninety new cases of eating disorders according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria were identified during the follow-up. In the multivariate logistic analysis, a higher risk of incident eating disorder was found for several exposures assessed at the beginning of follow-up, such as younger age, usually eating alone (odds ratio [OR]: 2.9; 95% confidence interval: 1.9-4.6), and frequently reading girls' magazines or listening to radio programs (OR: 2.1; 1.2-3.8 for those most frequently using both media). No independent association was found for television viewing or socioeconomic status. A marital status of parents different from "being married" was associated with a significantly higher risk in the multivariate analysis (OR: 2.0; 1.1-3.5). CONCLUSIONS: Our results support the role of mass media influences and parental marital status in the onset of eating disorders. The habit of eating alone should be considered as a warning sign of eating disorders. PMID- 12563058 TI - Genome scan for childhood and adolescent obesity in German families. AB - OBJECTIVE: Several genome scans have been performed for adult obesity. Because single formal genetic studies suggest a higher heritability of body weight in adolescence and because genes that influence body weight in adulthood might not be the same as those that are relevant in childhood and adolescence, we performed a whole genome scan. METHODS: The genome scan was based on 89 families with 2 or more obese children (sample 1). The mean age of the index patients was 13.63 +/- 2.75 years. A total of 369 individuals were initially genotyped for 437 microsatellite markers. A second sample of 76 families was genotyped using microsatellite markers that localize to regions for which maximum likelihood binomial logarithm of the odd (MLB LOD) scores on use of the concordant sibling pair approach exceeded 0.7 in sample 1. RESULTS: The regions with MLB LOD scores >0.7 were on chromosomes 1p32.3-p33, 2q37.1-q37.3, 4q21, 8p22, 9p21.3, 10p11.23, 11q11-q13.1, 14q24-ter, and 19p13-q12 in sample 1; MLB LOD scores on chromosomes 8p and 19q exceeded 1.5. In sample 2, MLB LOD scores of 0.68 and 0.71 were observed for chromosomes 10p11.23 and 11q13, respectively. CONCLUSION: We consider that several of the peaks identified in other scans also gave a signal in this scan as promising for ongoing pursuits to identify relevant genes. The genetic basis of childhood and adolescent obesity might not differ that much from adult obesity. PMID- 12563059 TI - Inadequate supervision as a cause of injury deaths among young children in Alaska and Louisiana. AB - OBJECTIVE: Inadequate supervision of children has contributed to injuries. However, the association of inadequate supervision with injury events in children has not been quantified. The purpose of this study was to describe and quantify the role of inadequate supervision of children in injury deaths. METHODS: Injury deaths among children aged 0 to 6 years in Alaska during 1993 to 1995 and Louisiana during 1994 were classified using 10 child safety standards to assess the role of parent/caregiver supervision in the circumstances of injury death. RESULTS: The leading categories of injury death for both states combined were motor vehicle injury and fire-related injury. Of the classifiable injury deaths in both states (157 [77%] of 203 deaths), the most commonly violated safety standard was "children should be supervised by a responsible care provider" (64 deaths [41%]). Of these deaths, the caregiver was absent in 38%, and the caregiver increased the danger to the child in 17%. Male injury deaths more typically involved a supervision standard violation. Drowning and pedestrian deaths typically involved a supervision standard violation, whereas asphyxiation, homicide, and occupant motor vehicle injury deaths did not. CONCLUSION: Alaska and Louisiana child injury deaths were mostly attributed to preventable violations of 10 child safety standards, most commonly the supervision standard. The methods in this report were useful in identifying target populations and causes of death, which can be used to plan and implement interventions to improve supervision of children. PMID- 12563060 TI - Physical fitness and C-reactive protein level in children and young adults: the Columbia University BioMarkers Study. AB - OBJECTIVE: To examine the association of physical fitness with C-reactive protein (CRP) level in children and young adults. METHODS: Subjects (N = 205) aged 6 to 24 years were enrolled in the Columbia University BioMarkers Study (1994-1998). Physical fitness was assessed using a non-effort-dependent treadmill testing protocol (physical work capacity at heart rate of 170 beats per minute). CRP level was measured using a high-sensitivity assay. RESULTS: Subjects were 54% female and 65% of Hispanic origin. Mean fitness level was higher in boys than in girls, but CRP levels did not differ by gender. Fitness level was inversely correlated with CRP (r = -0.22). This relationship was significant in boys (r = 0.32) but not in girls (r = -0.15). After multivariate regression adjustment for age, race/ethnicity, body mass index, and family history of early-onset ischemic heart disease, physical fitness remained inversely associated with CRP level in boys (beta = -0.02; standard error = 0.01). CONCLUSIONS: These findings indicate that physical fitness is inversely related to CRP level in children and that this relationship is more pronounced in boys than in girls. PMID- 12563061 TI - Can changes in clinical practice decrease the incidence of severe retinopathy of prematurity in very low birth weight infants? AB - OBJECTIVE: A wide variability in the incidence of severe retinopathy of prematurity (ROP) is reported by different centers. The altered regulation of vascular endothelial growth factor from repeated episodes of hyperoxia and hypoxia is 1 important factor in the pathogenesis of ROP. Strict management of O(2) delivery and monitoring to minimize these episodes may be associated with decreased rates of ROP. The objective of this study was to compare the incidence of and need for surgery for severe ROP (stages >or=3) in infants of 500 to 1500 g birth weight before and after the implementation of a new clinical practice of O(2) management in a large level 3 neonatal intensive care unit (NICU). METHODS: An oxygen management policy that included strict guidelines in the practices of increasing and weaning of fraction of inspired oxygen (FIO(2)) and the monitoring of O(2) saturation parameters in the delivery room, during in-house transport of infants to the NICU, and throughout hospitalization was implemented in April 1998. The main objectives were to monitor oxygenation levels more precisely and to avoid hyperoxia and repeated episodes of hypoxia-hyperoxia in very low birth weight infants. Included in the policy were equipment for monitoring, initiation of monitoring at birth, avoidance of repeated increases and decreases of the FIO(2), minimization of "titration" of FIO(2), modification of previously used alarm limits, and others. After an educational process, each staff member signed an agreement stating understanding of and future compliance with the guidelines. Examinations were performed by experienced ophthalmologists following international classification and American Academy of Pediatrics recommendations. ROP data from January 1997 to December 2002 for infants of 500 to 1500 g were analyzed as usual and also have been reported to Vermont Oxford Network since 1998. RESULTS: The incidence of ROP 3 to 4 at this center decreased consistently in a 5-year period from 12.5% in 1997 to 2.5% in 2001. The need for ROP laser treatment decreased from 4.5% in 1997 to 0% in the last 3 years. CONCLUSION: We observed a significant decrease in the rate of severe ROP in very low birth weight infants in association with an educational program provided to all NICU staff and the implementation and enforcement of clinical practices of O(2) management and monitoring. Although several confounders cannot be excluded, it is likely that differences in these clinical practices may be, at least in part, responsible for the documented intercenter variability in rates of ROP. PMID- 12563062 TI - Early versus late diagnosis: psychological impact on parents of children with cystic fibrosis. AB - OBJECTIVES: To explore the relationship between the period preceding diagnosis and the way parents of children with cystic fibrosis (CF) experience and handle their child's disease. DESIGN: A retrospective study. SETTING: CF Center "Noordwest Nederland," the Netherlands. PARTICIPANTS: Participants were the parents of children <13 years old with CF who were treated at the CF Center "Noordwest Nederland." The participants were divided into 2 groups according to the duration of prediagnostic period: <3 months (defined as early diagnosis) and >or=3 months (defined as late diagnosis). MAIN OUTCOME MEASURES: Experience of the prediagnostic period, contact with the medical profession, coping, future perspective, and attitudes toward neonatal screening for CF. RESULTS: Parents of 55 children were eligible for study participation; 45 were enrolled. Retrospectively, the period preceding an early diagnosis was less negatively experienced by parents than the period preceding a late diagnosis. Parents of children with an early diagnosis had retrospectively more confidence in the medical profession before confirmation of diagnosis. In general, parents in this study used adaptive coping styles. Duration of prediagnostic period was not significantly related to future perspective. Hopelessness seemed to be mainly determined by a severe course of disease as experienced by the parents. Ninety eight percent of all parents were in favor of neonatal screening for CF. CONCLUSIONS: A short prediagnostic period is associated with less negative feelings and increased confidence in the medical profession among parents of children with CF. Neonatal screening for CF may be of benefit to parents by removing the stress of a delayed diagnosis. PMID- 12563063 TI - Amplitude-integrated electroencephalography coupled with an early neurologic examination enhances prediction of term infants at risk for persistent encephalopathy. AB - OBJECTIVES: The objectives of this study were to determine, first, whether an early neurologic examination could predict a persistent abnormal neonatal neurologic state comparable to the amplitude-integrated electroencephalography (a EEG) and, second, whether a combination of the 2 methods would further enhance early identification of high-risk infants. METHODS: Fifty term infants were enrolled prospectively when they had evidence of intrapartum distress, Apgar score or=2 courses of oral glucocorticoids and were prescribed the same therapy for the index exacerbation. Children were considered unexposed when they had no exposure to oral glucocorticoids and were not prescribed any for the index exacerbation. Indices of bone metabolism were measured during the subsequent month. Cortisol responses to adrenocorticotrophic hormone stimulation and bone density were assessed 30 days after the index exacerbation. RESULTS: Eighty-three children (48 exposed, 35 unexposed) aged 2 to 17 years were enrolled. The median exposure level was 4 courses (range: 3-11) in the preceding year. Among exposed children, a transient decrease in serum osteocalcin was observed at the end of the 5-day course with a return to baseline by 30 days; no change was observed in urine pyridinoline cross links. Mean bone density z score was similar in the exposed (-0.61 +/- 1.0 [standard deviation]) and unexposed (-0.67 +/- 0.9) groups. No cases of abnormal response to adrenocorticotrophic hormone suggestive of adrenal insufficiency were documented in the exposed (95% confidence interval: 0%-7%) or unexposed (0%-10%) groups. CONCLUSIONS: Repeated short courses of oral glucocorticoids in the treatment of asthma seem to be reasonably safe; this practice was not associated with any lasting perturbation in bone metabolism, bone mineralization, or adrenal function. PMID- 12563068 TI - High rates of behavioral problems in perinatally HIV-infected children are not linked to HIV disease. AB - OBJECTIVE: Descriptive studies and clinical reports have suggested that human immunodeficiency virus (HIV)-positive children are at risk for behavioral problems. Inadequate control groups and sample sizes have limited the ability of investigators to consider multiple influences that place HIV-positive children at risk for poor behavioral outcomes. We examined the unique and combined influences of HIV, prenatal drug exposure, and environmental factors on behavior in children who were perinatally exposed to HIV. METHODS: Participants included 307 children who were born to HIV-positive mothers (96 HIV infected and 211 seroreverters) and enrolled in a natural history, longitudinal study of women to infant HIV transmission. Caregivers completed parent behavioral rating scales, beginning when the children were 3 years old. Data were also collected on prenatal drug exposure; child age, gender, and ethnicity; caregiver relationship to child; and birth complications. RESULTS: Multivariate analyses comparing the HIV-infected children with perinatally exposed but uninfected children from similar backgrounds failed to find an association between either HIV status or prenatal drug exposure and poor behavioral outcomes. The strongest correlates of increased behavioral symptoms were demographic characteristics. CONCLUSIONS: This study suggests that although a high prevalence of behavioral problems does exist among HIV-infected children, neither HIV infection nor prenatal drug exposure is the underlying cause. Rather, other biological and environmental factors are likely contributors toward poor behavioral outcomes. PMID- 12563069 TI - Availability of adolescent health services and confidentiality in primary care practices. AB - BACKGROUND: Little is known about availability of services and confidential care for adolescents in primary care practices or how availability among pediatric practices compares to that among other primary care practices. The objective of this study was to assess self-reported availability of services for medically emancipated conditions and confidential care in primary care practices, to compare physician responses to those from office staff who answer appointment lines, and to compare availability in pediatric practices to other primary care practice types. METHODS: We conducted a telephone survey of randomly selected practices from the Washington, DC, metropolitan area in pediatrics (Peds), internal medicine (IM), and family medicine (FM). We asked staff who answer appointment lines about availability of services for medically emancipated conditions and confidential appointments for adolescents. Physicians received the same questions via a mail survey. Responses from office staff and physicians in the same practice were linked for comparison. RESULTS: Of 434 practices contacted by telephone, 372 (86%) responded. Of the 615 physicians surveyed from these 372 practices, 264 (43%) from 170 practices responded to the mail survey. Peds practices were less likely than FM and IM practices to offer services for medically emancipated conditions and were less likely than FM practices to offer confidential services to adolescents. Office staff and physicians from FM and IM had higher agreement compared with Peds about availability of services for medically emancipated conditions. Agreement between office staff and physicians about provision of confidential appointments to adolescents was low among all practice types. However, having a written office policy on adolescent confidentiality was significantly associated with agreement between office staff and physicians about availability of confidential services. CONCLUSIONS: Care for medically emancipated conditions and confidential services for adolescents are limited among primary care practices, especially among pediatric practices. All primary care practice types had significant disagreement between office staff and physicians about availability of confidential services to adolescents. Adolescents who call appointment lines are likely to receive inaccurate information about confidentiality policies. Establishing written office policies on adolescent confidentiality may help to improve access to confidential care for adolescents. PMID- 12563070 TI - Adolescent participation in tobacco promotions: the role of psychosocial factors. AB - OBJECTIVES: To identify psychosocial factors that place adolescents at risk for participation in tobacco promotions, and to further investigate the hypothesis that psychosocial vulnerabilities have an indirect effect on smoking initiation among youth by way of involvement with tobacco promotions. METHODS: Data were from a follow-up telephone survey of youth in Massachusetts. A subset of adolescents who were not established smokers, had not smoked cigarettes in the past 30 days, and did not own a promotional item at baseline in 1993 was used for the analyses (n = 468). Bivariate analyses were used to evaluate the association between psychosocial vulnerabilities and subsequent acquisition of cigarette promotional items. Logistic regression was conducted to identify the set of factors that best predict attainment of tobacco promotional items, and to examine the mediating influence of item acquisition on the relation of between psychosocial vulnerabilities and smoking initiation. RESULTS: Adolescents who were academically disengaged at baseline were more likely to acquire a tobacco promotional item at follow-up. Academic disengagement was significantly associated with item acquisition, above and beyond the other psychosocial vulnerabilities. The direct effect of academic disengagement changed from marginally significant to nonsignificant when item acquisition was introduced. CONCLUSIONS: This study provides evidence that receptivity to tobacco promotional items is greatest among youth who are disengaged from school. PMID- 12563071 TI - Discovering fragile X syndrome: family experiences and perceptions. AB - We used surveys from 274 families who had at least 1 child with fragile X syndrome (FXS) to determine their experiences in discovering FXS, factors associated with the timeliness of discovery, and the perceived consequences of obtaining this information. For families of male children who were born in the last decade, someone first became concerned about the child's development at an average age of 13 months. Professional confirmation of a developmental delay did not occur until an average age of 21 months, and a FXS diagnosis occurred at an average age of nearly 32 months. Families reported several barriers to discovering FXS and frustration with the process. Many families had additional children with FXS without knowing reproductive risk. A range of perceived benefits and challenges associated with the discovery were reported. We conclude that selected pediatric practices could promote earlier identification but in only a limited way and predict that disorders such as FXS will continue to challenge current criteria for determining viable candidate disorders for newborn screening. PMID- 12563072 TI - Reducing errors in fluid therapy management. PMID- 12563073 TI - Scope of practice issues in the delivery of pediatric health care. AB - In recent years, there has been an increase in the number of nonphysician pediatric clinicians and an expansion in their respective scopes of practice. This raises critical public policy and child health advocacy concerns. The American Academy of Pediatrics (AAP) believes that optimal pediatric health care depends on a team-based approach with coordination by a physician leader, preferably a pediatrician. The pediatrician is uniquely suited to manage, coordinate, and supervise the entire spectrum of pediatric care, from diagnosis through all stages of treatment, in all practice settings. The AAP recognizes the valuable contributions of nonphysician clinicians, including nurse practitioners and physician assistants, in delivering optimal pediatric care. The AAP also believes that nonphysician clinicians who provide health care services in underserved areas should be supported by consulting pediatricians and other physicians using technologies including telemedicine. Pediatricians should serve as advocates for optimal pediatric care in state legislatures, public policy forums, and the media and should pursue opportunities to resolve scope of practice conflicts outside state legislatures. The AAP affirms that as nonphysician clinicians seek to expand their scopes of practice as providers of pediatric care, standards of education, training, examination, regulation, and patient care are needed to ensure patient safety and quality health care for all infants, children, adolescents, and young adults. PMID- 12563074 TI - Hearing assessment in infants and children: recommendations beyond neonatal screening. AB - Congenital or acquired hearing loss in infants and children has been linked with lifelong deficits in speech and language acquisition, poor academic performance, personal-social maladjustments, and emotional difficulties. Identification of hearing loss through neonatal hearing screening as well as objective hearing screening of all infants and children can prevent or reduce many of these adverse consequences. This report outlines the risk indicators for hearing loss, provides guidance for when and how to assess hearing loss, and addresses hearing referral resources for children of all ages. PMID- 12563075 TI - Heliox questions. PMID- 12563076 TI - Maternal report of prenatal alcohol use. PMID- 12563077 TI - Psychological maltreatment. PMID- 12563078 TI - Nosocomial enterococcal bacteremia in children. PMID- 12563079 TI - Discharge criteria for bronchiolitis patients. PMID- 12563080 TI - Ducks have ducks unless... PMID- 12563081 TI - Kawasaki syndrome hospitalizations among children in the United States, 1988 1997. PMID- 12563082 TI - "They're too smart for that": predicting what children would do in the presence of guns. AB - OBJECTIVES: We examined parents' beliefs about how children would react to finding guns, with particular emphasis on how parents reasoned about children's actions. METHODS: Based on a randomized telephone survey of Northeast Ohio residents, we focused on the 317 urban and 311 rural respondents who had children 5 to 15 years old in their homes. Respondents were asked about gun ownership and their expectations of how children would react to finding guns. Analysis examined responses in relation to various demographic and socioeconomic variables. RESULTS: All respondents--regardless of gun ownership, geography, race, gender, education level, income, or child age--were equally likely (approximately 87%) to believe that their children would not touch guns they found. Fifty-two percent of those reasoned that children were "too smart" or "knew better." Only 40% based their predictions on specific instructions they had given their children. Only 12% (15/122) of owners stored guns locked and unloaded. Only 3 of 13 variables tested were positively associated with safe storage: having a child 5 to 9 years old, having at least a 4-year college education, and having an income >or=65,000 dollars per year. CONCLUSION: Results indicate that parental beliefs may effectively relieve adults of responsibility and place the burden on children to protect themselves. The implication for injury prevention is that caregivers' unrealistic expectations of children's developmental levels and impulse control may influence storage decisions or the inclination to address gun safety issues with children or other adults with whom children spend time (relatives, playmates' parents). PMID- 12563083 TI - Childhood drowning: barriers surrounding private swimming pools. AB - OBJECTIVE: To investigate the causes of child drowning and determine the need for changes in the legislation as well as improvements to the inspection and enforcement of current legislation related to barriers surrounding private swimming pools. METHODS: There were 3 stages to the study: a retrospective review of coroner's data, an audit of swimming pool inspections, and in-depth interviews with swimming pool inspectors in Western Australia. The incidence of childhood drowning (per population) and compliance rates of swimming pools (per 1000 swimming pools) to the legislation were measured. RESULTS: During the 12-year observational period (1988-2000) 50 children younger than 5 years drowned in private swimming pools in Western Australia with an overall incidence of drowning of 4.4 per 100 000 children per year. Sixty-eight percent of drownings occurred in pools that did not have 4-sided fencing with an almost 2-fold increased risk (incidence rate ratio: 1.78; 95% confidence interval: 1.40-1.79) of a child's drowning in a swimming pool with 3-sided versus 4-sided fencing. The compliance rate of swimming pools (compliance to the current legislation) at first inspection was approximately 400 per 1000 swimming pools. CONCLUSIONS: Almost two thirds of the swimming pools in which children drowned had only 3-sided fencing. With a combination of a change in legislation, enhanced inspection processes, and public education, the incidence of drowning in private swimming pools in Western Australia could be reduced in the coming years. PMID- 12563084 TI - Examination of the communication practices between state newborn screening programs and the medical home. AB - BACKGROUND: As outlined in the Newborn Screening Task Force report published in August 2000, the newborn screening system is more than just testing, but also involves follow-up, diagnosis, treatment, and evaluation. As such, multiple professional and public partners need to be adequately involved in the system to help ensure success. In addition, newborn screening programs are state-based; therefore, policies and procedures vary from state to state. Historically, there has been little uniformity between state newborn screening programs. OBJECTIVE: To examine the communication practices of state newborn screening programs in the United States, particularly in relation to the medical home. METHODS: A facsimile survey of program staff in all US newborn screening programs. Survey data were collected in August 2000. RESULTS: All 51 programs participated. States were questioned about whether or not they had a procedure to identify the infant's medical home before the child's birth. Twelve states (24%) indicated that there was a procedure in place, whereas 39 states (76%) indicated that either no procedure existed or that they were unsure. In contrast, all state programs (except 1) indicated they notified the primary care physician about abnormal results and the need for follow-up. In addition, state programs reported that primary care physicians have responsibilities within the newborn screening system, particularly related to communicating with parents about screen-positive results and coordinating the collection of a second specimen. Thirty states reported that they directly notified parents of screen-positive infants of results and the need for follow-up as well. In regard to informing parents about newborn screening, 45% of states indicated that primary care physicians had some responsibility in informing parents about newborn screening. Most often, parents were informed about newborn screening just before specimen collection, and the most commonly used techniques to educate parents were informational brochures and conversation. Thirty-five states reported that they engaged in long-term tracking of infants after diagnosis confirmation. Only about half of these states provided long-term tracking of all of the conditions included in their state's newborn screening test panel. Of these 35 states that engaged in long-term tracking, 25 reported that they requested patient information from the primary care physician and/or subspecialist about ongoing treatment and follow-up. CONCLUSIONS: Newborn screening roles and responsibilities vary tremendously between states. Improvements in communication and better-defined protocols are needed, particularly between state newborn screening programs and the medical home. Many states identified the medical home as having significant responsibilities related to the short-term follow-up of screen-positive infants. Identification of the correct medical home before testing would help to reduce unnecessary time and frustration for state newborn screening programs, especially in the follow-up of infants that are difficult to locate. In addition, primary care physicians (ie, the medical home) need to have appropriate and ongoing involvement, including a mechanism to provide feedback to their state newborn screening program. This is particularly important given the adoption of tandem mass spectrometry by an increasing number of states, and the likely expansion of newborn screening in the future. Recommendations include the following: Primary care physicians should have appropriate and ongoing involvement in the newborn screening system and should be appropriately represented on state newborn screening advisory committees. States should develop protocols to identify the medical home before heelstick screening. States should work with families, primary care physicians, and prenatal health care professionals to develop well-defined systems for pretesting education of parents. All newborn screening results (both positive and negative) should be sent to the infant's medical home. If results are not received by the medical home, efforts should be made to obtain results. Medical homes and subspecialists should submit follow-up information on screen-positive infants and infants with confirmed diagnoses to the state newborn screening program, regardless of the existence of state requirements to do so, and efforts to build enhanced direct communication systems, linking state newborn screening programs to community-based medical homes, should continue. PMID- 12563085 TI - Racial disparity and modifiable risk factors among infants dying suddenly and unexpectedly. AB - BACKGROUND: Racial disparity in rates of death attributable to sudden infant death syndrome (SIDS) has been observed for many years. Despite decreased SIDS death rates following the "Back to Sleep" intervention in 1994, this disparity in death rates has increased. The prone sleep position, unsafe sleep surfaces, and sharing a sleep surface with others (bedsharing) increase the risk of sudden infant death. The race-specific prevalence of these modifiable risk factors in sudden unexpected infant deaths-including SIDS, accidental suffocation (AS), and cause of death undetermined (UD)-has not been investigated in a population-based study. Death rates attributable to AS and UD are also higher in African Americans (AAs) than in other races (non-AA). The potential contribution of unsafe sleep practices to this overall disparity in death rates is uncertain. OBJECTIVE: The objective of this study was to compare death rates attributable to SIDS and related causes of death (AS and UD) in AA and non-AA infants and the prevalence of unsafe sleep practices at time of death. Our hypothesis was that there is a large racial disparity in these modifiable risk factors at the time of death, and that public awareness of this could lead to improved intervention strategies to reduce the disparity in death rates. METHODS: In this population-based study, we retrospectively reviewed death-scene information and medical examiners' investigations of deaths in St Louis City and County between January 1, 1994, and December 31, 1997. The deaths of all infants <2 years old with the diagnoses of SIDS, AS, or UD were included. Sleep surfaces other than those specifically designed and approved for infant use were termed nonstandard (adult beds, sofas, etc). Denominators for our rate estimates were the number of births (AA and non AA) in St Louis City and County during the study period. RESULTS: The deaths of 119 infants were studied (81 AA and 38 non-AA). SIDS rates were much higher in AA than non-AA infants (2.08 vs 0.65 per 1000 live births), as was the rate of AS (0.47 vs 0.06). There was a trend for increased deaths diagnosed as UD in AA infants (0.36 vs 0.06). Bedsharing deaths were nearly twice as common in AAs (67.1% vs 35.1% of deaths), as were deaths on nonstandard sleep surfaces (79.0% vs 46.0%). Forty-nine percent (49.1%) of all infants who died while bedsharing were found on their backs or sides compared with 20.4% of infants who were not bedsharing. Overall, the fraction of infants found in these nonprone positions was not different for AA infants and non-AA infants (43.3% vs 38.5%). In AA and non-AA infants, factors that greatly increase the risk of bedsharing, such as sofa sharing or all-night bedsharing, were present in all or many bedsharing deaths. CONCLUSION: Among AA infants dying suddenly and unexpectedly, the high prevalence of nonstandard bed use and bedsharing may underlie, in part, their increased death rates. Public health messages tailored for the AA community have stressed first and foremost using nonprone sleep positions. The observation that there was no difference between AA and non-AA infants in position found at death suggests that racial disparity in sleep position is not the most important contributor to racial disparity in death rates. The finding that more infants died on their back or side while bedsharing than otherwise suggests that these sleep positions are less protective when associated with bedsharing. We conclude that public health information tailored for the AA community should give equal emphasis to risks and alternatives to bedsharing as to avoidance of the prone position. PMID- 12563086 TI - Review of multiple endocrine neoplasia type 2A in children: therapeutic results of early thyroidectomy and prognostic value of codon analysis. AB - OBJECTIVES: The aim of this study was first to investigate whether early total thyroidectomy (ETT; 1-5 years of age) can prevent medullary thyroid carcinoma with persistent or recurrent disease (PRD) in pediatric patients with multiple endocrine neoplasia type 2A (MEN-2A) and second, to evaluate the strength of codon analysis in children with MEN-2A as prognostic parameter. METHODS: Case reports and review of the literature for pediatric patients with MEN-2A were conducted. Inclusion criteria were age (0-20 years) and histologic degree of C cell disease (normal = N, C-cell hyperplasia = CCH, medullary thyroid carcinoma = MTC, metastatic MTC = MMTC). To evaluate therapeutic results of ETT (1-5 years) versus late total thyroidectomy (LTT; 6-20 years), age-dependent histologic stages of C-cell disease and postoperative occurrence of PRD were compared. Prognostic value of specific codons, age-dependent histologic distribution, and long-term outcome were analyzed. RESULTS: In a total of 260 cases, 42 (16%) underwent ETT, and 218 (84%) underwent LTT. Histologic analysis showed significant difference between ETT versus LTT (57% vs 76%) regarding malignant stage of C-cell disease (of combined rate of MTC and MMTC). Long-term outcome was documented in 74 patients (28%). During a median follow-up period of 2 years (range: 0-15 years), 21 of 65 of the LTT group versus 0 of 9 of the ETT group suffered PRD. Information about codon analysis was available in 150 patients (58%). Mutated codons were c634 (63%), c618 (19%), c620 (9%), and c804 (6%). Codon-related histologic analysis resulted in prognostic differences: 81% of patients with c634-mutation had MCT or MMTC in contrast to c804 (44%), c618 (34%), and c620 (7%). Fifteen of 17 MMTC and 7 of 9 PRD occurred in patients with c634-mutation. CONCLUSIONS: 1) ETT until 5 years of age in MEN-2A gene carriers results in significant reduction of MTC and MMTC in favor of CCH and improved disease-free long-term outcome. 2) Codon analysis is an important prognostic factor. Timing of TT could be individualized based on codon-specific prognosis. Until more detailed knowledge is available, consequent genetic and biochemical screening is mandatory for appropriate individual timing of ETT before age of 5 years. PMID- 12563087 TI - Expression of CD40 ligand on CD4+ T-cells and platelets correlated to the coronary artery lesion and disease progress in Kawasaki disease. AB - OBJECTIVE: Kawasaki disease (KD) is an acute febrile vasculitic syndrome in children. CD40 ligand (CD40L) has been implicated in certain types of vasculitis. We proposed that CD40L expression might be correlated with coronary artery lesions in KD. METHODS: Blood samples were collected from 43 patients with KD before intravenous immunoglobulin (IVIG) treatment and 3 days afterward. Forty three age-matched febrile children with various diseases were studied in parallel as controls. CD40L expression on T-cells and platelets were detected by flow cytometry, and soluble CD40L (sCD40L) levels were measured by enzyme-linked immunosorbent assay. RESULTS: We found that CD40L expression on CD4(+) T-cells was significantly higher in patients with KD than in the febrile control (FC) group (28.69 +/- 1.17% vs 4.37 +/- 0.36%). CD40L expression decreased significantly 3 days after IVIG administration (28.69 +/- 1.17% vs 13.53 +/- 0.55%). CD40L expression on platelets from patients with KD was also significantly higher than in the FC group (8.20 +/- 0.41% vs 1.26 +/- 0.12%) and decreased after IVIG therapy. sCD40L levels were also significantly higher in KD patients with those of FC (9.69 +/- 0.45 ng/mL vs 2.25 +/- 0.19 ng/mL) but were not affected by IVIG treatment 3 days afterward (9.69 +/- 0.45 ng/mL vs 9.03 +/- 0.32 ng/mL). More interesting, we found that in KD patients, CD40L expression on CD4(+) T-cells and platelets but not on CD8(+) T-cells or sCD40L was correlated with the occurrence of coronary artery lesions. CONCLUSIONS: CD40L might play a role in the immunopathogenesis of KD. IVIG therapy might downregulate CD40L expression, resulting in decrease of CD40L-mediated vascular damage in KD. This implicates that modulation of CD40L expression may benefit to treat KD vasculitis. PMID- 12563088 TI - Effect of exercise on immunologic factors in breast milk. AB - OBJECTIVE: Although it is well documented that breast milk provides optimal nutrition and immune benefits to the infant, factors that influence the immunologic composition of breast milk are less understood. A recent study reported that immunoglobulin A (IgA) levels in breast milk are lower after exercise compared with resting concentrations. However, the women exercised until exhaustion. The effect of moderate exercise on immunologic components in breast milk has not been reported. Therefore, the purpose of this study was to 1) compare the levels of immunologic compounds in breast milk of exercising women with the milk of sedentary women and 2) determine whether 30 minutes of moderate exercise affects immunologic properties of breast milk. METHODS: Exclusively lactating women were studied at 3 months' postpartum. Women in the exercise group (EG; n = 29) reported exercising aerobically at least 30 minutes/d for 3 days/wk, and women in the sedentary group (SG; n = 24) had exercised once a week or less during the previous 6 weeks. Cardiovascular fitness levels and concentrations of IgA, lactoferrin, and lysozyme in milk were measured. A subsample of the EG (n = 17) participated in a 30-minute exercise session at 75% of maximum heart rate and a rest session of 30 minutes of sitting rest on 2 separate days. Breast milk samples were collected before and 10 and 60 minutes after exercise and rest sessions. IgA, lactoferrin, and lysozyme concentrations were measured. RESULTS: Women in the EG had a higher level of cardiovascular fitness than women in the SG (39.7 +/- 1.0 vs 32.4 +/- 1.0 mL O(2)/kg/min). Milk concentrations of IgA, lactoferrin, or lysozyme were not significantly different between groups. In addition, there were no significant differences in the concentrations of IgA, lactoferrin, or lysozyme after moderate exercise compared with sitting rest. CONCLUSION: Moderate exercise during lactation improves cardiovascular fitness without affecting levels of IgA, lactoferrin, or lysozyme in breast milk. PMID- 12563089 TI - Temperament and physical performance in children with osteogenesis imperfecta. AB - OBJECTIVE: Children with osteogenesis imperfecta (OI) must participate in therapy to achieve motor performance objectives. Their behavioral style may influence motor performance. For this reason, the temperament of children with types III or IV OI was assessed prospectively to 1) compare their temperament with that of nondisabled children, 2) investigate the relationship between temperament and gross motor performance, and 3) examine relationships among temperament, parental overprotection and coping, physical activity, muscle strength, and motor performance. METHODS: Age-appropriate Carey Temperament Scales, Brief Assessment of Motor Function (BAMF), and the Vulnerable Child/Overprotecting Parents Scale were completed for 35 children 1 to 12 years old. Additional measures included the Childhood Health Assessment Questionnaire, Parent Daily Hassles Scale, manual muscle testing, Pediatric Activity Record, and a Summed Severity Score. Spearman correlations and multiple regression were used to identify and predict significant relationships. RESULTS: Temperament of children with OI differed from age-based norms in only 1 domain: activity. Motor performance (BAMF) correlated significantly with 3 domains of temperament: persistence (r = -.48), approach (r = -.34), and activity (r =.40). Activity was also related to the ratio of head circumference to body length (r = -.45) and the number of fractures in the preceding year (r = -.35). Parents' reports of their daily hassles significantly correlated with several domains of the child's temperament. No significant relationships were identified between parental overprotection and temperament or motor performance. CONCLUSIONS: The temperament of children with types III and IV OI does not differ from that of their nondisabled peers, with the exception of lower activity scores. Although it is considered a biological attribute, the expression of temperament, specifically activity, may be influenced by learned behaviors. Because gross motor performance is related to activity, persistence, and approach/avoidance, knowledge of an individual's temperament may enhance the child's ability to benefit from interventions to improve motor skill and activity levels. PMID- 12563090 TI - Tobacco counseling at well-child and tobacco-influenced illness visits: opportunities for improvement. AB - OBJECTIVE: To assess the frequency of clinician-reported delivery of counseling for avoidance of child environmental tobacco smoke (ETS) exposure and tobacco use at periodic well-child visits and at illness visits for asthma and otitis media (OM). METHODS: Combined data from the National Ambulatory Medical Care Survey and the outpatient portion of the National Hospital Ambulatory Medical Care Survey from 1997 to 1999 were analyzed. The frequency of pediatric visits (25% (95% confidence interval: 1.2-13.9) and remained statistically significant after adjustment for treatment. When we compared the PE-positive group with the acquired immunodeficiency syndrome (AIDS)/PE-negative group (children who developed clinical category C and without neurologic manifestations) in a cross sectional study within 12 months before PE or AIDS diagnosis, respectively, the %CD8(+) T-lymphocytes were significantly lower in the PE-positive group. Normalized absolute counts of CD8(+) T-lymphocytes with respect to seroreverting children were significantly lower in the group of children with encephalopathy with respect to the AIDS/PE-negative group (data not shown). It is interesting that a statistically significant increase was observed in circulating monocyte percentages and absolute counts shortly before the first neurologic symptoms compared with values after PE was established and with those from HIV-1-infected controls. With respect to AIDS-related events, PE was strongly associated with anemia and lymphoid interstitial pneumonitis in the PE-positive group with respect to a group of children with AIDS but without PE. CONCLUSION: HIV-1 infection of the central nervous system (CNS) remains an important clinical concern. The first step toward PE prevention in HIV-1-infected children should be directed at predicting risk of PE and thus the prompt and reliable identification of infants who are at risk for CNS disease progression. Low blood CD8(+) T lymphocytes is a strong early predictive marker of PE emergence in vertical HIV-1 infection. Indeed, among all of the immunologic and virologic variables assessed in this observational study, the only significant difference during the first months of life are the CD8(+) T-lymphocytes. A peak of significantly higher peripheral monocytes before the onset of PE with respect to established PE has not been previously described, and strengthens the growing evidence that an increased traffic of monocytes to the brain may be a key factor in triggering neurologic symptoms. The suppression of HIV-1 replication is dependent on the presence of a relatively small number of HIV-1-specifof HIV-1-specific CD8(+) T lymphocytes, and it is possible that the duration of the neurologically asymptomatic phase for any given child may depend mostly on the magnitude of specific CD8(+) T-lymphocyte responses. Thus, a decrease of CD8(+) T-lymphocytes would diminish the host capacity to control viral infection, as reported in animal models, enabling infected macrophages to cross the blood-brain barrier. Our results advocate the use of CD8(+) T-lymphocyte and monocyte counts to follow up HIV-1-infected children. We suggest that CD8(+) T-lymphocytes may be the nexus for many different aspects of the disease, namely loss of control of HIV-1 replication determining higher VL, increased traffic of activated and/or infected monocytes, spread of infection to immune sanctuaries, and finally clinical neurologic emergence of PE. Moreover, we suggest that CD8(+) T-lymphocytes or/and monocytes may be used as putative biological markers of neuropathogenicity. This might suggest their use in decision making of when to start more effective antiretroviral regimens for HIV-1 infection of the CNS and the need of new therapies either to preserve or to augment an adequate CD8(+) T-lymphocyte immune response. Early detection of children who are at risk for developing PE is particularly important because aggressive highly active antiretroviral therapy improves neurologic symptoms, allows possible use of neuroprotective treatment to prevent further development of encephalopathy, and emphasizes the relevance of developing therapies aimed to enhance CD8(+) T-lymphocyte function. In conclusion, the surrogate markers routinely used in clinical practice for HIV-1 infection (ie, CD4(+) T-lymphocyte counts and VL) seem to be insufficient to evaluate the clinical involvement of the CNS. Other systemic markers, as the recent proposed markers for PE evolution (cerebrospinal fluid VL by lumbar puncture and brain atrophy by cerebral magnetic resonance imaging) are undoubtedly more invasive than measuring CD8(+) T-lymphocyte and monocyte counts, when the neurologic manifestations of PE are still preventable. PMID- 12563092 TI - Infectious disease hospitalizations among American Indian and Alaska native infants. AB - OBJECTIVE: To describe the burden and trends in hospitalizations associated with infectious diseases among American Indian and Alaska Native (AI/AN) infants. METHODS: First-listed infectious disease hospitalizations and hospitalization rates among AI/AN infants and infants in the general US population from 1988-1999 were analyzed by using Indian Health Service/tribal hospital discharge data and the National Hospital Discharge Survey data, respectively. RESULTS: Infectious disease hospitalizations accounted for 53% of all AI/AN infant hospitalizations and approximately 43% of all US infant hospitalizations during 1988-1999. The annual hospitalization rate for infectious diseases among AI/AN infants declined from 27,486 per 100,000 infants in 1988 to 14,178 per 100,000 infants in 1999. However, the rates for AI/AN infants within the Alaska, Southwest, and Northern Plains regions remained higher than that for the general US infant population at the end of the study period. Lower respiratory tract infection hospitalizations accounted for almost 75% of AI/AN infant infectious disease hospitalizations, and the lower respiratory tract infection hospitalization rate for AI/AN infants was twice that for US infants. CONCLUSIONS: Although infectious disease hospitalization rates for AI/AN infants have declined, AI/AN infants continue to have a higher infectious disease burden than the general US infant population. PMID- 12563093 TI - Myocardial infarction and transient ventricular dysfunction in an adolescent with sickle cell disease. AB - We report a case of an adolescent who had sickle cell disease and previous evidence of myocardial damage and presented with abdominal pain and rapid progression to cardiogenic shock and subsequent development of myocardial infarction. To our knowledge, this represents only the second report of a case of acute myocardial ischemia and subsequent infarction resulting transient ventricular dysfunction reported in a child with sickle cell disease successfully treated with exchange transfusion. The pathophysiology of this complication remains unclear, and cardiac complications may remain undetected as lung, bone, and brain infarcts are more common and the pain associated with sickle cell crisis may mask the ischemic symptoms. Multiple factors may contribute to ischemia in addition to the presence of a vaso-occlusive crisis or infection. Acute or chronic myocardial ischemia are probably more prevalent than currently known. PMID- 12563094 TI - Alteplase in the treatment of complicated parapneumonic effusion: a case report. AB - The treatment of complex parapneumonic effusions in children remains controversial, with some advocating less invasive, strictly medical management and others supporting a more aggressive approach of thoracotomy with or without decortication. Recent advances, including video-assisted thoracoscopic surgery and intrapleural fibrinolytic therapy, offer new options for effective treatment. We report the first case of successful resolution of a complex parapneumonic effusion in a 16-month-old girl with the use of tissue plasminogen activator (alteplase), infused via a catheter in the pleural space. PMID- 12563095 TI - Accidental scald burns in sinks. AB - Scald burns to the feet and lower extremities in children are described in the literature as often resulting from forced immersions. This report illustrates 3 cases of burns whose distribution and historical factors identify them as accidental. The location of these accidental burns is similar to those found in inflicted injury, but the patterns were indicative of flowing water burns, not forced immersions. Burns in these locations may be confused with abuse. Medical providers need to be aware of information that may enable them to distinguish the 2 causes. Effective caregiver education regarding the importance of lowering the temperature of water heaters and discouraging play in household sinks is critical to prevent additional tap water scald burn injuries. PMID- 12563096 TI - Septic arthritis in an infant with vesicoureteral reflux and urinary tract infection. AB - A 4-week-old boy with previous urinary tract infection and documented vesicoureteral reflux presented with urosepsis and septic arthritis of the right hip. Compliance with prophylactic antibiotic therapy had been poor at home. Complications such as bone and joint infection are known to occur after urinary tract infection in children with urologic abnormalities. However, previous similar reports describe discovery of the urinary tract anomalies only as part of an evaluation performed after the systemic complications have occurred. The purpose of this report is to stress the importance of defining urinary tract abnormalities in a case of antenatal hydronephrosis or at the time of the first urinary tract infection in infants so that appropriate investigations, management, and support of parental compliance can be undertaken to avoid systemic complications. PMID- 12563098 TI - Hormesis and toxicological risk assessment. PMID- 12563099 TI - Chemical allergy: considerations for the practical application of cytokine profiling. AB - Chemical respiratory allergy is an important occupational health problem, but there are currently available no validated methods for hazard identification. This is due in part to the fact that the relevant cellular and molecular mechanisms of sensitization of the respiratory tract have been unclear, with particular controversy regarding the role of IgE. There is now increasing evidence that respiratory sensitization is associated with the preferential activation of type 2 T lymphocytes and the expression of type 2 cytokines interleukin (IL)-4, IL-5, IL-10, and IL-13. Type 2 cell products favor immediate type hypersensitivity reactions, serving as growth and differentiation factors for mast cells and eosinophils, the cellular effectors of the clinical manifestations of the allergic responses, and promoting IgE antibody production. There has been considerable interest in the application of cytokine profiling for the characterization of chemical allergens, with cytokine phenotypes analyzed in freshly isolated tissue, or following culture in the presence or absence of mitogen at the level of protein secretion or mRNA expression. Experience to date suggests that the measurement of induced cytokine secretion profiles shows promise for the hazard identification and characterization of chemical respiratory allergens. The purpose of this brief review article is to consider the approaches available and to highlight key procedural issues. PMID- 12563100 TI - Disruption of gap junctions in toxicity and carcinogenicity. AB - Although the specific role of connexin-mediated gap junctional intercellular communication in the control of cell homeostasis, proliferation, and death are still not clear, several lines of evidence support these roles. The disturbance of this communication, through multiple mechanisms, may in the short term be a protective mechanism to limit the spread of toxicity in a tissue following chemical or radiation damage. However, sustained downregulation confers a loss of tumor-suppressive action. Consequently, connexin dysfunction has been associated with both the action of many carcinogens and being a feature of cancer per se. Connexins offer not only a target for cancer chemoprevention but also for exploitation in chemotherapy through the "bystander" effect. PMID- 12563101 TI - Cadmium pathways during gestation and lactation in control versus metallothoinein 1,2-knockout mice. AB - Effects of metallothionein (MT) on cadmium absorption and transfer pathways during gestation and lactation in mice were investigated. Female 129/SvJ metallothionein-knockout (MT1,2KO) and metallothionein-normal (MTN) mice received drinking water containing trace amounts of (109)CdCl(2) (0.15 ng Cd/ml; 0.074 micro Ci (109)Cd/ml). (109)Cd and MT in maternal, fetal, and pup tissues were measured on gestation days 7, 14, and 17 and lactation day 11. In dams, MT influenced both the amount of (109)Cd transferred from intestine into body (two- to three-fold higher in MT1,2KO than MTN dams) and tissue-specific (109)Cd distribution (higher liver/kidney ratio in MT1,2KO dams). Placental (109)Cd concentrations in MT1,2KO dams were three- and seven-fold higher on gestation days 14 and 17, respectively, than in MTN dams. Fetal (109)Cd levels were low in both mouse types, but at least 10-fold lower in MTN fetuses. MT had no effect on the amount of (109)Cd transferred to pups via milk; furthermore, 85-90% of total pup (109)Cd was recovered in gastrointestinal tracts of both types, despite high duodenal MT only in MTN pups. A relatively large percentage of milk-derived intestinal (109)Cd was transferred to other pup tissues in both MT1,2KO and MTN pups (14 and 10%, respectively). These results demonstrate that specific sequestration of cadmium by both maternal and neonatal intestinal tract does not require MT. Although MT decreased oral cadmium transfer from intestine to body tissues at low cadmium exposure levels, MT did not play a major role in restricting transfer of cadmium from dam to fetus via placenta and to neonate via milk. PMID- 12563102 TI - Comparison of the hemoglobin adducts formed by administration of N methylolacrylamide and acrylamide to rats. AB - Acrylamide (AM) and N-methylolacrylamide (NMA) are used in the formulation of grouting materials. AM undergoes metabolism to a reactive epoxide, glycidamide (GA). Both AM and GA react with hemoglobin to form adducts that can be related to exposure to AM. The objective of this study was to evaluate the extent to which NMA could form the same adducts as AM. N-(2-carbamoylethyl)valine (AAVal derived from AM) and N-(2-carbamoyl-2-hydroxyethyl)valine (GAVal derived from GA) were measured following a single oral dose of AM (50 mg/kg) or NMA (71 mg/kg) in male F344 rats. AAVal and GAVal were measured by a modified Edman degradation to produce phenylthiohydantoin derivatives and liquid chromatography/tandem mass spectrometry. In AM-treated rats, AAVal was 21 +/- 1.7-pmol/mg globin (mean +/- SD, n = 4), and GAVal was 7.9 +/- 0.8 pmol/mg. In NMA-treated rats, AAVal was 41 +/- 4.9 pmol/mg, and GAVal was 1.4 +/- 0.1 pmol/mg. Whether AAVal was derived from reaction of NMA with globin followed by loss of the hydroxymethyl group, or loss of the hydroxymethyl group to form AM prior to reaction with globin, is not known. However, the higher ratio of AAVal:GAVal in NMA-treated rats (29 vs. 2.6 in AM-treated rats) suggests that reaction of NMA with globin is the predominant route to AAVal in NMA-treated rats. The detection of GAVal in NMA-treated rats indicates oxidation of NMA, either directly or following conversion to AM. The lower levels of GAVal on NMA administration suggest that a much lower level of epoxide was formed than compared with AM treatment. PMID- 12563103 TI - Mode of action and pharmacokinetic studies of 2-butoxyethanol in the mouse with an emphasis on forestomach dosimetry. AB - Chronic inhalation studies with 2-butoxyethanol (BE) conducted by the National Toxicology Program identified the forestomach and liver of B6C3F1 mice as target organs for tumorigenicity (NTP, 2000). Previous studies have shown that the liver tumors likely resulted from chronic hemolysis-induced oxidative stress. For the forestomach lesions seen in mice, chronic contact irritation (cytotoxicity) and regenerative hyperplasia are hypothesized to result in forestomach tumor development. To test this hypothesis, several experiments were conducted to address the sensitivity of the mouse forestomach to BE administered by various routes. Oral administration of undiluted BE was shown to cause irritation and a compensatory proliferative response in the mouse forestomach, confirming that direct contact between the forestomach and BE, which can occur via grooming of BE condensed on the fur during inhalation exposures, can cause irritation. However, only small amounts of BE (<10 mg/kg) were detected on the fur of mice at the end of 6-h, whole-body or nose-only inhalation exposures to the highest concentration used in the NTP chronic inhalation studies (250 ppm). Furthermore, no significant differences were detected in the end-exposure blood concentrations of BE and butoxyacetic acid (BAA) between these types of exposures. In addition, parenteral administration of BE (ip and sc injection) also resulted in forestomach lesions, indicating that there may be sources other than grooming for BE- or BAA-induced forestomach irritation. In the pharmacokinetic study, BE and, to a lesser extent, BAA was eliminated more slowly from the forestomach tissue of mice than from blood or other tissues, following either oral gavage or ip injection. The forestomach was the only tissue with detectable levels of BE at 24 h. BE and BAA were both excreted in the saliva and were present in stomach contents for a prolonged period of time following these routes of exposure, which may further contribute to forestomach tissue dosimetry. Thus, there appear to be multiple mechanisms behind the increased levels of BE and BAA in the forestomach tissue of mice, which together can contribute to a prolonged contact irritation, compensatory hyperplasia, and tumorigenicity in mice. The relevance of these effects in humans, who lack a forestomach, is questionable. PMID- 12563104 TI - Inhibition of mouse hepatocyte gap junctional intercellular communication by phenobarbital correlates with strain-specific hepatocarcinogenesis. AB - The inhibition of gap junctional intercellular communication (GJIC) is a common effect of nongenotoxic carcinogens and might be a biomarker for these agents. To further test this relationship, we hypothesized that phenobarbital would inhibit mouse hepatocyte GJIC and this would correlate with strain-specific hepatocarcinogenicity. Phenobarbital is a strong nongenotoxic hepatocarcinogen in B6C3F1 mice, but not in C57BL/6 mice. Hepatocytes were isolated from males of both strains, placed in coculture with rat liver epithelial cells, and treated with phenobarbital for up to 14 days. Male mice were also administered PB by single intraperitoneal injection (0.1 mg/kg), then sacrificed 24 h later, or given phenobarbital in the drinking water (500 ppm) for 14 days before sacrifice. GJIC was assayed in cocultures by fluorescent dye microinjection and in isolated liver tissue by fluorescent dye "cut-loading." Phenobarbital decreased GJIC only in cultured B6C3F1 hepatocytes; this was dose-responsive and temporary, because hepatocyte GJIC returned to control levels within 24 h of phenobarbital exposure. Administration of phenobarbital to mice for 14 days also decreased hepatocyte dye coupling in B6C3F1 liver, but this effect was not seen in C57BL/6 mice or observed after a single administration of the drug. Phenobarbital did not alter connexin32 and connexin26 expression, but increased hepatic Cyp2b1 expression and the liver weight:body weight ratio in both strains. In summary, phenobarbital inhibited mouse hepatocyte GJIC in vivo and in vitro and in correlation with strain-specific hepatocarcinogenicity. These data support the hypothesis that decreased GJIC is a biomarker for nongenotoxic carcinogens and involved in their carcinogenic mechanism. PMID- 12563105 TI - Toxicology and carcinogenesis studies of microencapsulated citral in rats and mice. AB - Citral, a widely used natural ingredient, is added to foods and cosmetics as a flavoring and fragrance agent. Male and female F344/N rats and B6C3F1 mice were exposed to microencapsulated citral in the feed for 14 weeks or two years. All studies included untreated and vehicle control groups. In the 14-week studies, rats and mice were given diets containing 3,900, 7,800, 15,600, or 31,300 ppm citral. In rats, food consumption was reduced in the two highest dose groups. In mice an apparent increase in food consumption was observed, but was due to mice scattering the feed. Body weights of all treated animals were less than controls. All rats and four male mice were killed moribund in the high dose groups. In rats, forestomach and kidney lesions were observed. At the higher doses, lesions observed in the bone marrow, testes, and thymus in rats and in the ovary in mice were considered related to inanition and resultant moribundity. In the two-year studies, rats were exposed to 1,000, 2,000, or 4,000 ppm citral. Body weights were reduced in the 4,000 ppm rats. Mice were exposed to 500, 1,000, or 2,000 ppm citral. Body weights in the 1,000 and 2,000 ppm groups were reduced. No neoplasms were attributed to citral in rats or mice. Malignant lymphoma occurred with a positive trend and was significantly greater than controls in female mice in the 2,000 ppm group. However, the incidences were within the NTP historical control range and could not be clearly related to citral administration. PMID- 12563106 TI - Respiratory exposure to diesel exhaust particles decreases the spleen IgM response to a T cell-dependent antigen in female B6C3F1 mice. AB - We investigated the systemic immunotoxic potential of respiratory exposure to diesel exhaust particles (DEP) in this study. Female B6C3F1 mice (approximately 8 weeks old) were exposed to increasing concentrations of DEP intratracheally, 3 times every two weeks, and sacrificed 2 or 4 weeks after the first exposure. The systemic toxicity and immune status in mice were evaluated. Mice exposed to DEP (1 to 15 mg/kg) showed no significant changes in body, spleen, or liver weights. Lung weights were increased in the mice exposed to 15 mg/kg DEP for 2 or 4 weeks. Except for a decreased platelet count, no significant alterations occurred in hematological parameters following DEP exposure. The number of splenic anti-sheep red blood cell (sRBC) IgM antibody-forming cells (AFC) decreased following DEP exposure for 2 weeks. This effect was less severe following 4 weeks of exposure and was only evident in the high dose group. Exposure to DEP also resulted in a significant decrease in the absolute numbers and the percentages of total spleen cells for total, CD4(+), and CD8(+) T cells, while the numbers of B cells and total nucleated cells in spleen were not significantly changed. The proliferative response of splenocytes to the T-cell mitogen, concanavalin A (ConA), as well as their production of IL-2 and IFN-gamma, was decreased dose-dependently following exposure of mice to DEP for 2 weeks, whereas proliferation was not changed in response to anti-CD3 monoclonal antibody. In summary, short-term respiratory exposure of mice to DEP resulted in systemic immunosuppression with evidence of T cell-mediated and possibly macrophage-mediated mechanisms. PMID- 12563107 TI - Significance of 2-methoxypropionic acid formed from beta-propylene glycol monomethyl ether: integration of pharmacokinetic and developmental toxicity assessments in rabbits. AB - Commercial grade propylene glycol monomethyl ether (PGME), which is composed of > 99.5% alpha-isomer and < 0.5% beta-isomer, has been shown in several studies to have a low potential for developmental toxicity. Nonetheless, questions have been raised about potential human developmental toxicity due to beta-PGME, because it can be metabolized to 2-methoxypropionic acid (MPA), a compound bearing structural similarity to the teratogen, methoxyacetic acid (MAA). Accordingly, a series of in vivo developmental toxicity, whole embryo culture, and in vivo pharmacokinetic experiments were conducted in New Zealand White rabbits (highly sensitive to these compounds) to better understand the developmental toxicity potential of MPA and the kinetics of its formation from beta-PGME. For the in vivo developmental toxicity studies, groups of 20 inseminated rabbits were gavaged with 0, 10, 26, or 78 mg/kg/day of MPA on gestation day (GD) 7-19, followed by fetal evaluation on GD 28. Results with MPA were compared with those of rabbits similarly dosed with 0, 2.5, 7.5, or 15 mg/kg/day of MAA. Developmental toxicity no-observable-effect levels (NOEL) were approximately 10 fold higher for MPA (26 mg/kg/day) than for MAA (2.5 mg/kg/day). Also, the severity of effects caused by MPA was less than that of MAA, and unlike MAA, MPA was not selectively toxic to the fetus. This differential toxicity was also seen in whole embryo cultures of GD 9 rabbit embryos, in which there were no adverse effects of MPA (1.0, 5.0 mM) or its parent compound, beta-PGME (0.5, 2.0 mM), but severe dysmorphogenesis in 100% of embryos cultured in 5.0 mM MAA. The pharmacokinetics study showed rapid and complete conversion of beta-PGME to MPA, with a relatively long elimination half-life (33-44 h) for MPA. However, peak and AUC concentrations of MPA in blood associated with the MPA LOEL dose of 78 mg/kg/day were 1.3 mM and 52.9 mM-h/l, respectively, suggesting a relatively high threshold based on internal dosimetry. Taken together, these data indicate a negligible risk of developmental toxicity due to MPA formation from the small amounts of beta-isomer present in commercial PGME. PMID- 12563108 TI - 3-methylindole-induced toxicity to human bronchial epithelial cell lines. AB - Transfected BEAS-2B cells that express different cytochrome P450 enzymes were used to assess whether human bronchial epithelial cell lines are target cells for 3-methylindole (3MI)-induced damage. Four different transfected BEAS-2B lines overexpressing P450s 2A6, 3A4, 2F1, and 2E1 (B-CMV2A6, B-CMV3A4, B-CMV2F1, and B CMV2E1), respectively, were compared. The B-CMV2F1 and B-CMV3A4 cells were the most susceptible to 3MI-mediated cytotoxicity, measured by leakage of lactate dehydrogenase into the medium after a 48-h incubation. The toxicity was ameliorated by pretreatment with 1-aminobenzotriazole (ABT). Depletion of glutathione with diethylmaleate decreased the onset and increased the extent of cell death with 3MI. Thus, 3MI is cytotoxic to immortalized bronchial epithelial cells overexpressing 2F1 without concomitant depletion of GSH, but depletion of GSH modestly enhances the cytotoxicity of 3MI to human lung cells. Additional studies clearly demonstrated that a low concentration of 3MI (10 micro M) induced apoptosis in BEAS-2B cells that was measured by DNA fragmentation, and apoptosis was inhibited by the presence of ABT. The B-CMV2F1 cells overexpressing 2F1 demonstrated increased apoptosis (measured by Annexin-V binding) at 24 h with 100 micro M 3MI. Therefore, CYP2F1 in human bronchial epithelial lung cells may bioactivate 3MI to 3-methyleneindolenine, which induces programmed cell death at relatively low concentrations. Human lung cells may be susceptible to this prototypical pneumotoxicant. PMID- 12563109 TI - Inhaled environmental combustion particles cause myocardial injury in the Wistar Kyoto rat. AB - Epidemiologists have associated particulate matter (PM) air pollution with cardiovascular morbidity and premature mortality worldwide. However, experimental evidence demonstrating causality and pathogenesis of particulate matter (PM) induced cardiovascular damage has been insufficient. We hypothesized that protracted, repeated inhalation by rats of oil combustion-derived, fugitive emission PM (EPM), similar in metal composition to selected sources of urban air PM, causes exposure duration- and dose-dependent myocardial injury in susceptible rat strains. Zinc was the only primary water-leachable/bioavailable element of this EPM. Male Sprague-Dawley (SD), Wistar Kyoto (WKY), and spontaneously hypertensive (SH) rats were exposed nose-only to EPM (2, 5, or 10 mg/m(3), 6 h/day for 4 consecutive days or 10 mg/m(3), 6 h/day, 1 day/week for 4 or 16 consecutive weeks). Two days following the last EPM exposure, cardiac and pulmonary tissues were examined histologically. The results showed that particle laden alveolar macrophages were the only pulmonary lesions observed in all three rat strains. However, WKY rats exposed to EPM (10 mg/m(3) 6 h/day, 1 day/week for 16 weeks) demonstrated cardiac lesions with inflammation and degeneration. To further characterize the nature of EPM-associated lesions, more rigorous histopathological and histochemical techniques were employed for WKY and SD rats. We examined the hearts for myocardial degeneration, inflammation, fibrosis, calcium deposits, apoptosis, and the presence of mast cells. Decreased numbers of granulated mast cells, and multifocal myocardial degeneration, chronic-active inflammation, and fibrosis were present in 5 of 6 WKY rats exposed to EPM for 16 weeks. None of these lesions were present in WKY exposed to clean air. EPM related cardiac lesions were indistinguishable from air-exposed controls in SD and SH rats. This study demonstrates that long-term inhalation exposures to environmentally relevant PM containing bioavailable zinc can cause myocardial injury in sensitive rats. These findings provide supportive evidence for the epidemiological associations of cardiovascular morbidity and ambient PM. PMID- 12563110 TI - The hormetic dose-response model is more common than the threshold model in toxicology. AB - The threshold dose-response model is widely viewed as the most dominant model in toxicology. The present study was designed to test the validity of the threshold model by assessing the responses of doses below the toxicological NOAEL (no observed adverse effect level) in relationship to the control response (i.e., unexposed group). Nearly 1,800 doses below the NOAEL, from 664 dose-response relationships derived from a previously published database that satisfied a priori entry criteria, were evaluated. While the threshold model predicts a 1:1 ratio of responses "greater than" to "less than" the control response (i.e., a random distribution), a 2.5:1 ratio (i.e., 1171:464) was observed, reflecting 31% more responses above the control value than expected (p < 0.0001). The mean response (calculated as % control response) of doses below the NOAEL was 115.0% +/- 1.5 standard error of the mean (SEM). These findings challenge the long standing belief in the primacy of the threshold model in toxicology (and other areas of biology involving dose-response relationships) and provide strong support for the hormetic-like biphasic dose-response model characterized by a low dose stimulation and a high-dose inhibition. These findings may affect numerous aspects of toxicological and biological/biomedical research related to dose response relationships, including study design, risk assessment, as well as chemotherapeutic strategies. PMID- 12563111 TI - The combined effects of vinclozolin and procymidone do not deviate from expected additivity in vitro and in vivo. AB - The combination effects of the well-known antiandrogenic fungicides, vinclozolin and procymidone, were tested both in vitro and in vivo. In vitro both vinclozolin and procymidone significantly inhibited the binding of agonist to the androgen receptor with the concentration that resulted in 50% inhibition (IC(50)) values of 0.1 and 0.6 micro M, respectively. By applying the isobole method, the effect of combining the two pesticides in vitro was found to be additive. In castrated testosterone-treated rats the administration of vinclozolin starting at 10 mg/kg led to a decrease in organ weight of all tested reproductive organs. The levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were increased significantly with doses of 100 mg/kg vinclozolin and above. Expression of the androgen-responsive gene, TRPM-2, was increased starting at 100 mg/kg vinclozolin. For procymidone, reproductive organ weights were diminished at 10 mg/kg and LH was increased at a concentration of 25 mg/kg and above, compared to the testosterone-treated controls. FSH was significantly increased only at 25 mg/kg procymidone. The studied gene expressions were changed by 100 mg/kg procymidone. Dosing the animals with a combination of a 1:1 mixture of vinclozolin and procymidone resulted in a weight reduction in the reproductive organs and an increase of serum LH and FSH as early as with 10 mg/kg combined dose. The relative expressions of TRPM-2 and PBP C3 were changed compared to controls at 100 mg/kg. The level of 5-HT in the rat brain was increased after a dose of 10 mg/kg. Using the isobole method, comparisons of the observed and predicted effects assuming additivity on reproductive organ weights, hormone levels, and gene expression showed agreement and thus the combination effects are suggested to be additive in vivo as well as in vitro. PMID- 12563112 TI - Effects of chronic intrathecal infusion of a partial differential opioid agonist in dogs. AB - To define the effects of chronic spinal exposure to a highly selective partial differential opioid agonist c[DPen(2),DPen(5)]enkephalin (DPDPE), adult beagles were prepared with chronic lumbar intrathecal catheters. Groups of dogs received intrathecal infusions (100 micro l/h) of saline (vehicle), DPDPE 3 mg/ml or 6 mg/ml for 28 days. Over the 28-day period, saline or 3 mg/ml showed minimal changes in neurological function, whereas in the 6 mg/ml animals, prominent hind limb dysfunction evolved over the 28-day interval. Histopathology in control animals displayed a modest pericatheter reaction considered normal for this model. Dogs receiving DPDPE (three of four at 6 mg/ml and one of four at 3 mg/ml) but not saline (zero of four) developed large inflammatory masses (granulomas) in the intrathecal space located proximal to the catheter tip. In these masses, severe chronic inflammatory changes in combination with necrosis and fibrosis was detected. Occasional focal destruction of neuropil was detected also in the adjacent spinal cord parenchyma. These masses contained extensive accumulation of mouse antihuman macrophages (MAC)-positive inflammatory cells expressing tumor necrosis factor-alpha (TNF-alpha), revealing infiltration of macrophages, granulocytes, and monocytes. In separate animals, prepared with dual intrathecal catheters, lumbar CSF was sampled at specified time points following intrathecal bolus (3 mg/ml) and 24 h DPDPE infusion (3 mg/ml and 6 mg/ml). Steady-state cerebrospinal fluid (CSF) DPDPE levels were 18.6 +/- 1.0 and 22.6 +/- 4.0 micro g/ml for 3 mg/ml and 6 mg/ml infusions respectively. These results indicate that this partial differential opioid agonist DPDPE produces a concentration and time dependent formation of an intrathecal inflammatory mass. PMID- 12563113 TI - Cyclosporin A induced internalization of the bile salt export pump in isolated rat hepatocyte couplets. AB - Isolated rat hepatocyte couplets were used to perform the comparative study of two widely used immunosuppressors, cyclosporin A (CsA) and tacrolimus (FK506) on hepatocanalicular function. We assessed canalicular function by counting the percentage of couplets that were able to accumulate the fluorescent cholephile, cholyl-lysyl-fluorescein (CLF), into the canalicular vacuole between the two cells, i.e., canalicular vacuole accumulation (CVA) of CLF. Compared to controls (DMSO-treated cells), CsA, in the approximate range of concentrations used therapeutically, caused inhibition of CVA of CLF, disorganization of the bile salt export pump (Bsep) localization at canalicular level resulting in its relocation into the cell, and disruption of the pericanalicular F-actin cytoskeleton. In contrast, FK506, at both approximately therapeutic and supratherapeutic concentrations, had no deleterious effect upon CVA of CLF, upon the localization of the bile salt transporter at the canalicular membrane, or on the organization of the pericanalicular F-actin cytoskeleton. These results point to transporter and cytoskeletal disorganization as contributors or determinants of CsA-induced cholestasis at canalicular level, whereas FK506 does not appear to produce these cholestasis-determining responses even at supratherapeutic concentrations. PMID- 12563115 TI - Agree that idiosyncratic hepatotoxicity in TRO-treated patients is uncertain, but concerned about the data. PMID- 12563116 TI - Measurement variability and confidence intervals in medicine: why should radiologists care? AB - In radiology, appropriate diagnoses are often based on quantitative data. However, these data contain inherent variability. Radiologists often see P values in the literature but are less familiar with other ways of reporting statistics. Statistics such as the SD and standard error of the mean (SEM) are commonly used in radiology, whereas the CI is not often used. Because the SEM is smaller than the SD, it is often inappropriately used in order to make the variability of the data look tighter. However, unlike the SD, which quantifies the variability of the actual data for a single sample, the SEM represents the precision for an estimated mean of a general population taken from many sample means. Since readers are usually interested in knowing about the variability of the single sample, the SD often is the preferred statistic. Statistical calculations combine sample size and variability (ie, the SD) to generate a CI for a population proportion or population mean. CIs enable researchers to estimate population values without having data from all members of the population. In most cases, CIs are based on a 95% confidence level. The advantage of CIs over significance tests (P values) is that the CIs shift the interpretation from a qualitative judgment about the role of chance to a quantitative estimation of the biologic measure of effect. Proper understanding and use of these fundamental statistics and their calculations will allow more reliable analysis, interpretation, and communication of clinical information among health care providers and between these providers and their patients. PMID- 12563117 TI - RSNA Board restructures cabinet to strengthen support of radiologic science and research. PMID- 12563119 TI - Patient-doctor relationships. PMID- 12563120 TI - The RSNA Editorial Fellowship: editorial fellows' perspective. PMID- 12563121 TI - Superior semicircular canal dehiscence syndrome and multi-detector row CT. PMID- 12563122 TI - Neuroimaging and early diagnosis of Alzheimer disease: a look to the future. AB - Alzheimer disease (AD), a progressive neurodegenerative disorder, is the most common cause of dementia in the elderly. Current consensus statements have emphasized the need for early recognition and the fact that a diagnosis of AD can be made with high accuracy by using clinical, neuropsychologic, and imaging assessments. Magnetic resonance (MR) or computed tomographic (CT) imaging is recommended for the routine evaluation of AD. Coronal MR images can be useful to document or quantify atrophy of the hippocampus and entorhinal cortex, both of which occur early in the disease process. Both volumetric and subtraction MR techniques can be used to quantify and monitor dementia progression and rates of regional atrophy. MR measures are also increasingly being used to monitor treatment effects in clinical trials of cognitive enhancers and antidementia agents. Positron emission tomography (PET) and single photon emission CT offer value in the differential diagnosis of AD from other cortical and subcortical dementias and may also offer prognostic value. In addition, PET studies have demonstrated that subtle abnormalities may be apparent in the prodromal stages of AD and in subjects who carry susceptibility genes. PET ligands are in late-stage development for demonstration of amyloid plaques, and human studies have already begun. Functional MR-based memory challenge tests are in development as well. PMID- 12563123 TI - CT evaluation of bone dehiscence of the superior semicircular canal as a cause of sound- and/or pressure-induced vertigo. AB - PURPOSE: To describe the computed tomographic (CT) findings at different collimation widths associated with superior semicircular canal (SSC) dehiscence syndrome and to determine the frequency of these findings in a control population. MATERIALS AND METHODS: Temporal bone CT scans with 1.0-mm and/or 0.5 mm collimation were obtained in 50 patients with sound- and/or pressure-induced vestibular symptoms. The control population consisted of 50 patients undergoing CT at 1.0-mm collimation and 57 patients undergoing CT at 0.5-mm collimation for other reasons. RESULTS: SSC dehiscence was documented on CT scans in all 36 patients with the clinical syndrome, with bilateral findings in six patients. Six other patients without specific clinical signs appeared to have dehiscence on 1.0 mm-collimated scans. Intact bone overlaying the SSC was subsequently identified with 0.5-mm-collimated CT in each case. On the 1.0-mm-collimated scans in 50 control patients, an area judged as possible or definite dehiscence was identified in 18 of 100 ears. The bone overlaying the SSC was intact in each of the 114 control ears evaluated with 0.5-mm-collimated CT. CT findings from the patients with vestibular symptoms combined with those in the control population indicated that the positive predictive value of an apparent dehiscence in the diagnosis of SSC dehiscence syndrome improved from 50% with 1.0-mm-collimated CT with transverse and coronal images to 93% with 0.5-mm-collimated CT with reformation in the plane of the SSC. CONCLUSION: The positive predictive value of CT in identification of SSC dehiscence syndrome improves with 0.5-mm-collimated helical CT and reformation in the SSC plane. PMID- 12563124 TI - Salivary gland tumors: diagnostic value of gadolinium-enhanced dynamic MR imaging with histopathologic correlation. AB - PURPOSE: To evaluate the diagnostic value of gadolinium-enhanced dynamic magnetic resonance (MR) imaging of salivary gland tumors and correlate the MR imaging and histopathologic findings. MATERIALS AND METHODS: Thirty-three salivary gland tumors in 29 patients were examined preoperatively at gadolinium-enhanced dynamic MR imaging. There were 22 benign and 11 malignant tumors. Dynamic contrast material-enhanced MR images were obtained for 5 minutes. Time of peak enhancement (T(peak)) and washout ratio (WR) were determined from time-signal intensity curves (TICs). Microvessel count and cellularity-stromal grade were evaluated histopathologically. The strengths of correlations between T(peak) and microvessel count and between WR and cellularity-stromal grade were statistically analyzed. Statistical analysis was also performed to determine whether any differences among the various histopathologic tumor types existed [corrected]. In a validation study, 13 salivary gland tumors in 13 patients were examined consecutively. RESULTS: At a T(peak) of 120 seconds, malignant tumors could be differentiated from pleomorphic adenomas but not from Warthin tumors. A WR of 30%, however, enabled differentiation between malignant and Warthin tumors. Classification of TICs on the basis of a T(peak) of 120 seconds and a WR of 30% had high sensitivity (91%) and specificity (91%) in the differentiation of benign and malignant tumors. Correlations between T(peak) and microvessel count (P <.0001, rho = -0.800) and between WR and cellularity-stromal grade (P =.0105, rho = 0.572) were significant. The validation study also yielded high sensitivity (100%) and specificity (80%) in the differentiation between benign and malignant tumors. CONCLUSION: Gadolinium-enhanced dynamic MR imaging is useful for differentiating benign from malignant salivary gland tumors. PMID- 12563125 TI - Optic neuritis: evaluation with orbital Doppler sonography. AB - PURPOSE: To evaluate orbital blood flow velocities with Doppler sonography in patients with acute unilateral optic neuritis. MATERIALS AND METHODS: Orbital Doppler sonography was performed in 20 patients with acute unilateral optic neuritis. Optic neuritis was diagnosed by a neurologist on the basis of clinical presentation, presence of decreased visual acuity, and assessment of visual evoked potentials. The peak systolic and end diastolic velocities and the resistive index were measured in the ophthalmic and central retinal arteries of both orbits. The values obtained from affected and unaffected orbits were compared by using the paired t test. RESULTS: The peak systolic and end diastolic velocities in the ophthalmic artery were significantly increased in the affected orbits (for peak systolic velocity P <.001, for end diastolic velocity P <.05). Resistive indexes in the ophthalmic arteries did not differ (P >.05). The difference between the peak systolic and end diastolic velocities and resistive indexes in the central retinal arteries of affected and normal eyes was not statistically significant (P >.05). CONCLUSION: Peak systolic and end diastolic velocities in the ophthalmic artery are increased in patients with acute optic neuritis. PMID- 12563126 TI - Posterior pituitary bright spot in large adenomas: MR assessment of its disappearance or relocation along the stalk. AB - PURPOSE: To evaluate the disappearance or relocation patterns of posterior pituitary bright spot (PPBS) in the infundibuloneurohypophyseal (INH) system and endocrinologic implications in large pituitary adenomas. MATERIALS AND METHODS: Sixty-nine patients with adenoma and supradiaphragmatic extension were classified into PPBS-visible and PPBS-nonvisible groups on the basis of findings at preoperative T1-weighted magnetic resonance (MR) imaging. The adenoma shapes were classified into hourglass type with indentation and barrel type without indentation at the diaphragmatic level. RESULTS: The PPBS-visible group included 55 (80%) patients. PPBS most commonly occurred at the distal pituitary stalk immediately above the diaphragm in 48 patients with hourglass-type adenoma. In the remaining seven patients with barrel-type adenoma, PPBS occurred in the sella or in varying sites along the pituitary stalk. Postoperatively, two patients, whose PPBS became nonvisible, developed persistent diabetes insipidus. The PPBS nonvisible group included 14 (20%) patients. Five had hourglass-type and nine had barrel-type adenoma. Occurrence of the barrel type was marked. In these patients, four developed postoperative permanent diabetes insipidus. CONCLUSION: The diaphragm, a probable major anatomic determinant of indentation, may serve as a transportation blockade and facilitate proximal accumulation of PPBS material, as evidenced in the hourglass-type adenoma. PPBS was more commonly nonnvisible in the barrel-type adenoma. The presence of PPBS in the INH system indicates its functional maintenance in large adenomas. PMID- 12563127 TI - Vertebral compression fractures: pain reduction and improvement in functional mobility after percutaneous polymethylmethacrylate vertebroplasty retrospective report of 245 cases. AB - PURPOSE: To describe the immediate outcome of a large cohort of patients who underwent percutaneous polymethylmethacrylate (PMMA) vertebroplasty for treatment of one or more vertebral fractures. MATERIALS AND METHODS: This retrospective cohort study included seven university-based and private hospitals in the United States. Of 488 consecutive patients (mean age, 76 years) who underwent percutaneous PMMA vertebroplasty between 1996 and 1999, 245 were successfully interviewed retrospectively after vertebroplasty (median time, 7 months). Through telephone interview, patients completed our self-developed questionnaire designed to measure pain (10-point scale), ambulation (five-point scale), and ability to perform activities of daily living (ADL) (five-point scale) before and after vertebroplasty. Differences in reported pain, ambulation, and ability to perform ADL before and after vertebroplasty were evaluated with paired t tests. Differences in proportions were compared with the McNemar test. Subgroup analyses were performed to assess the consistency of differences in pre- and postprocedural pain and functional status by patient age, number of fractures, time from fracture to vertebroplasty, and time from vertebroplasty to questionnaire completion. RESULTS: On a 10-point scale, mean pain decreased from 8.9 before vertebroplasty to 3.4 afterward (P <.001). Seventy-two percent of patients had substantially impaired ambulation before vertebroplasty compared with 28% afterward (P <.001). Ability to perform ADL was also significantly improved following vertebroplasty (P <.001). Twelve patients (4.9%) experienced symptomatic complications (none major or life threatening). CONCLUSION: Treatment of vertebral fractures with percutaneous PMMA vertebroplasty appears to be safe and results in substantial immediate pain reduction and improved functional status. A randomized controlled trial appears warranted to assess the efficacy and safety of vertebroplasty. PMID- 12563128 TI - Osteoarthritis: MR imaging findings in different stages of disease and correlation with clinical findings. AB - PURPOSE: To determine whether knee pain, stiffness, and limited function in patients with different stages of osteoarthritis correlate with the degree of disease assessed on magnetic resonance (MR) images and radiographs. MATERIALS AND METHODS: Radiographs in 50 patients with varying degrees of osteoarthritis of the knee were assessed by using the the Western Ontario and McMaster University (WOMAC) osteoarthritis index and the Kellgren-Lawrence (KL) scale. MR images were obtained and analyzed by two readers for cartilage lesions, bone marrow edema pattern, and ligamentous and meniscal lesions. RESULTS: Thirteen of 16 knees with a KL score of 4 showed full-thickness cartilage lesions and bone marrow edema pattern. Cruciate ligament tears were found in five of 12 knees with a KL score of 3 and in nine of 16 knees with a KL score of 4. While the KL score correlated significantly (P <.05) with the grade of cartilage lesions, and a substantially higher percentage of lesions with higher KL scores were found on MR images, the correlations between MR imaging findings and KL score versus clinical findings were not significant (P >.05). Significant differences between WOMAC scores were found only for the grades of cartilage lesions (P <.05). CONCLUSION: Cartilage lesions, bone marrow edema pattern, and meniscal and ligamentous lesions were frequently demonstrated on MR images in patients with advanced osteoarthritis. Clinical findings showed no significant correlations with KL score and extent of findings at MR imaging. PMID- 12563129 TI - Cartilage lesions in the hip: diagnostic effectiveness of MR arthrography. AB - PURPOSE: To evaluate the diagnostic performance of magnetic resonance (MR) arthrography in the detection of articular cartilage lesions in patients suspected of having femoroacetabular impingement and/or labral abnormalities. MATERIALS AND METHODS: Forty-two MR arthrograms obtained in 40 patients with a clinical diagnosis of femoroacetabular impingement and/or labral defect were retrospectively analyzed. Two readers independently interpreted the images for cartilage lesion location, depiction, and characteristics. Within 6 months after MR arthrography, each patient underwent open hip surgery, during which the entire cartilage of the hip joint was inspected. Sensitivity, specificity, accuracy, and positive and negative predictive values were calculated. kappa values were calculated to quantify the level of interobserver agreement. RESULTS: At surgery, most (37 [88%] of 42) cartilage defects were identified in the anterosuperior part of the acetabulum. In 23 (55%), 12 (29%), 10 (24%), and 10 (24%) hips, lesions were found in the posterosuperior acetabulum, anteroinferior acetabulum, posteroinferior acetabulum, and femoral head, respectively. The sensitivities and specificities of MR arthrographic detection of cartilage damage in all regions combined were 79% (73 of 92 regions) and 77% (91 of 118 regions), respectively, for reader 1 and 50% (46 of 92 regions) and 84% (99 of 118 regions), respectively, for reader 2. At interobserver comparison, agreement was fair (kappa = 0.31) for detection of cartilage lesions in the femoral head and poor (kappa 3.0 cm) tumors, presence of a tumor component in the renal sinus was a significant negative predictor of technical success (P =.004); only five of these 11 tumors were completely treated, compared with 11 of 11 tumors without a renal sinus component. A similar analysis was not possible for small tumors because no small tumors involved the renal sinus. Four complications occurred in a total of 54 ablation sessions: one minor hemorrhage, two major hemorrhages, and one ureteral stricture. CONCLUSION: RF ablation of RCC can be successful in exophytic RCC tumors up to 5.0 cm in size. Tumors larger than 3.0 cm with a component in the renal sinus are more difficult to treat but can be ablated successfully. PMID- 12563136 TI - Uterine fibroids: uterine artery embolization versus abdominal hysterectomy for treatment--a prospective, randomized, and controlled clinical trial. AB - PURPOSE: To evaluate the effectiveness of uterine artery embolization (UAE) in the management of bleeding in patients with uterine fibroids and to compare UAE with hysterectomy, particularly with regard to length of hospital stay and associated complications (ie, safety). MATERIALS AND METHODS: A prospective clinical trial was performed with patients who were randomly assigned to one of two groups: patients who were offered the option of undergoing either UAE or hysterectomy (group 1) and patients who were not informed of the alternative treatment-that is, UAE (group 2). The primary variables that were considered for evaluation of the effectiveness, efficiency, and safety of the two procedures were, respectively, bleeding cessation, total length of hospital stay, and resulting complications. The lengths of hospital stay in the two study arms were compared on an intent-to-treat basis. Owing to crossover between the treatment arms, however, effectiveness and safety were evaluated on the basis of the actual treatment received. RESULTS: The clinical success rate for the patients who underwent UAE, which was based on the cessation of bleeding, was 86% (31 of 36 patients). The mean hospital stay for group 1 was 4.14 days shorter than that for group 2 (P <.001). Ten (25%) of the 40 patients who underwent UAE experienced minor complications, in contrast to four (20%) of the 20 who underwent hysterectomy and experienced major complications. CONCLUSION: Compared with hysterectomy, UAE is safe and effective for treatment of bleeding fibroids, necessitates a shorter hospital stay, and results in fewer major complications. PMID- 12563137 TI - Recurrent traumatic urethral strictures near the external sphincter: treatment with a covered, retrievable, expandable nitinol stent--initial results. AB - PURPOSE: To investigate the clinical effectiveness of a covered, retrievable, expandable nitinol stent in treating traumatic urethral strictures near the external sphincter. MATERIALS AND METHODS: The stent was 10 mm in diameter when fully expanded and 40-50 mm long. To make it removable, two nylon drawstrings were attached to its lower inner margin. Twelve consecutive men (19-67 years; mean age, 47 years) with traumatic urethral strictures near the external sphincter that were refractory to endoscopic urethrotomy were treated. With fluoroscopic guidance, a stent was placed to completely bridge the external sphincter. The stent was electively removed with a retrieval hook wire 2 months after placement. Stent placement and removal were successful in all patients, with no procedural complications. RESULTS: Mean maximum urine flow rate was 5 mL/sec (range, 3-7 mL/sec) before stent placement and 27 mL/sec (range, 16-40 mL/sec) at 1 week after placement. During the mean follow-up of 20 months (range, 2-37 months) after the first stent removal, strictures recurred in eight of 12 patients. For the eight patients with recurrence, a second stent was placed and 4 months later was removed from seven of the eight patients. During the mean follow up of 18 months (range, 4-32 months) after the second stent removal, strictures recurred in two patients, in whom a third stent was placed and then removed 4 months later, with good results. Four (33%) of 12 patients, five (62%) of eight patients, and two (100%) of two patients were successfully treated with placement of the first stent, the second stent, and the third stent, respectively. In the remaining patient, the second stent remains in place. CONCLUSION: Placement of a covered, retrievable, expandable nitinol stent seems to be effective in treating urethral strictures near the external sphincter and warrants further investigation. PMID- 12563138 TI - Treatment of focal liver tumors with percutaneous radio-frequency ablation: complications encountered in a multicenter study. AB - PURPOSE: To report complications encountered by members of a collaborative group who performed radio-frequency (RF) ablation in patients with focal liver cancer. MATERIALS AND METHODS: Members of 41 Italian centers that were part of a collaborative group used a percutaneous internally cooled RF ablation technique and a standardized protocol for follow-up. They completed a questionnaire regarding number of deaths, presumed cause of death, and likelihood of its relationship to the RF procedure; number and types of major complications; and types of minor complications and side effects. Enrollment included 2,320 patients with 3,554 lesions (size, 3.1 cm +/- 1.1 [SD] in diameter): 1,610 had hepatocellular carcinoma with chronic liver disease; 693 had metastases, predominantly from colorectal cancer (n = 501); and 17 had cholangiocellular carcinoma. Number and characteristics of complications (ie, deaths and major and minor complications) attributed to the procedure were reported. Data were subsequently analyzed with analysis of variance to determine whether the major complication rate was related to tumor size, number of ablation sessions, or electrode type (single or cluster). RESULTS: In total, 3,554 lesions were treated. Six deaths (0.3%) were noted, including two caused by multiorgan failure following intestinal perforation; one case each of septic shock following Staphylococcus aureus-caused peritonitis, massive hemorrhage following tumor rupture, liver failure following stenosis of right bile duct; and one case of sudden death of unknown cause 3 days after the procedure. Fifty (2.2%) patients had additional major complications. The most frequent of these were peritoneal hemorrhage, neoplastic seeding, intrahepatic abscesses, and intestinal perforation. An increased number of RF sessions were related to a higher rate of major complications (P <.01), whereas the number of complications was not significantly different when tumor size or electrode type were compared. Minor complications were observed in less than 5% of patients. CONCLUSION: Results of this study confirm that RF ablation is a relatively low-risk procedure for the treatment of focal liver tumors. PMID- 12563139 TI - Femoral pseudoaneurysms: management with percutaneous thrombin injections- success rates and effects on systemic coagulation. AB - PURPOSE: To determine the success rate of percutaneous ultrasonographically (US) guided thrombin injection in the treatment of femoral pseudoaneurysms and to evaluate the effects of thrombin injection on systemic coagulation parameters. MATERIALS AND METHODS: Fifty femoral pseudoaneurysms (37 simple pseudoaneurysms with one lobe and 13 complex pseudoaneurysms with two or three lobes) were treated with US-guided percutaneous thrombin injections. Pseudoaneurysm size, neck length and width, thrombin dose, outcome of therapy, and complications were documented prospectively. Duplex sonographic follow-up examinations were performed at 12-24 hours and 5-7 and 21-25 days. In 25 patients, activated thromboplastin time, Quick test (prothrombin time), thrombin time, fibrinogen, D dimer, antithrombin III, thrombin-antithrombin III complex, and prothrombin fragments 1 and 2 were determined before and at 2, 5, and 10 minutes after thrombin injection. Differences in results before and those after thrombin injection were evaluated by means of the one-sample t test. RESULTS: Mean volume of pseudoaneurysms was 5.84 cm(3) +/- 4.89 (SD). Fifty-eight thrombin injections were performed. Mean thrombin dose was 357 IU +/- 291 in simple and 638 IU +/- 549 in complex pseudoaneurysms. Primary success rate was 36 of 37 (97%) for simple and eight of 13 (61%) for complex pseudoaneurysms. Reperfusion occurred in four complex pseudoaneurysms (none in simple ones). Secondary success rate was 100%. No thromboembolic, infectious, or allergic complications occurred. During follow-up, reperfusion was detected in one patient with a complex pseudoaneurysm. Levels of thrombin-antithrombin III complex increased significantly (P <.05) after thrombin injection, whereas changes in all other laboratory tests were not significant. CONCLUSION: US-guided percutaneous injection of thrombin is successful and safe in the management of femoral pseudoaneurysms. The increase of thrombin-antithrombin III complex indicates the possibility of thrombin passage into the arterial circulation. PMID- 12563140 TI - MR image-guided endovascular procedures with the ultrasmall superparamagnetic iron oxide SH U 555 C as an intravascular contrast agent: study in pigs. AB - PURPOSE: To evaluate the feasibility of using the ultrasmall superparamagnetic iron oxide (USPIO) SH U 555 C as an intravascular contrast agent for magnetic resonance (MR) image-guided vascular procedures with an open MR imaging system. MATERIALS AND METHODS: All experiments were performed with MR imaging at 0.2 T. MR image-guided interventions were performed in USPIO-enhanced vessels in four pigs. With near real-time MR image guidance (acquisition time, 0.64 second per section), the splenic and renal arteries were consecutively catheterized by using a susceptibility artifact-based catheter-guide wire combination. Angioplasty and stent implantation were performed four times in the renal artery and twice in the iliac artery. Intraaortal signal intensity (SI) was measured during the interventions. RESULTS: After administration of SH U 555 C (40 micromol of iron per kilogram of body weight), a three-dimensional MR angiographic sequence was performed that allowed visualization of the abdominal and pelvic vessels that were as small as 2 mm in diameter. Catheterization, angioplasty, and stent implantation were successfully guided in the USPIO-enhanced vasculature. Sixty minutes after contrast agent injection, the mean aortic SI was 70% of the maximum measured enhancement levels. CONCLUSION: One intravenous injection of SH U 555 C enabled long, continuous intravascular SI enhancement at MR angiography, and, in combination with susceptibility artifact-based device tracking, the injection allowed the performance of MR imaging-guided intravascular interventions in an open MR imaging system. PMID- 12563142 TI - Endovascular stents in pulmonary valve and artery in swine: feasibility study of MR imaging-guided deployment and postinterventional assessment. AB - PURPOSE: To assess the feasibility of using magnetic resonance (MR) imaging to guide stent deployment in the pulmonary valve and artery and evaluate, after stent deployment, the position and morphology of and blood flow through the stent. MATERIALS AND METHODS: Angiography and 1.5-T MR imaging were performed in a dual-imaging suite. Nitinol stents were placed in the pulmonary valve and main pulmonary artery in five pigs by using MR imaging guidance. For interactive MR imaging monitoring of catheter manipulation and stent delivery, balanced fast field-echo and T1-weighted turbo field-echo sequences were used. Visualization of the delivery system was based on T2* (with air as the contrast material) or T1 (with gadodiamide as the contrast material). After stent deployment, the position and morphology of and flow through the stent were verified with multiphase multisection balanced fast field-echo and velocity-encoded cine MR imaging. Findings at angiography and postmortem examination also helped verify stent placement. The paired Student t test was used for data analysis. RESULTS: The stent was successfully deployed in all animals. The stent was placed distal to the pulmonary valve in four animals and across the pulmonary valve in one animal. The position and morphology of the stent were clearly depicted on balanced fast field-echo images. In the animal with the stent placed across the pulmonary valve, the pulmonary regurgitant fraction was 37%; this was not seen in the animals with stents placed distal to the pulmonary valve. No complication (eg, stent migration, intramural injury, or vascular perforation) was noted during the intervention. Findings at angiography and postmortem examination confirmed the position of the stents. CONCLUSION: MR imaging has the potential to guide stent placement in the pulmonary valve or artery and to evaluate flow volume within the stent lumen after the intervention. PMID- 12563141 TI - Imaging with 99mTc ECDG targeted at the multifunctional glucose transport system: feasibility study with rodents. AB - PURPOSE: To evaluate the feasibility of technetium 99m ((99m)Tc) ethylenedicysteine-deoxyglucose (ECDG) imaging in tumor-bearing rodents. MATERIALS AND METHODS: ECDG was synthesized by means of reacting ethylenedicysteine with glucosamine, with carbodiimide as the coupling agent. Hexokinase assays were performed at an ultraviolet wavelength of 340 nm. To determine whether blood glucose level could be altered, ECDG or glucosamine was injected into six rats. In a separate study, ECDG followed by insulin was administered to three rats. To determine biodistribution, lung tumor cells were intramuscularly injected into the hind legs of 18 nude mice. The animals were then injected with (99m)Tc ECDG or fluorine 18 ((18)F) fluorodeoxyglucose (FDG) (0.037-0.074 MBq per mouse). Radioactivity was measured in tissue excised from the animals. Scintigraphy was performed in three groups: in group 1 to demonstrate that different-sized tumors could be imaged after (99m)Tc ECDG administration, in group 2 to ascertain whether tumor uptake of (99m)Tc ECDG was perfusion related, and in group 3 to demonstrate that tumor uptake of (99m)Tc ECDG occurred by means of a glucose-mediated process. RESULTS: ECDG was positive for phosphorylation at hexokinase assay. Blood glucose level increased with ECDG injection and decreased with insulin administration. Tumor-to-brain tissue and tumor-to-muscle tissue ratios of (99m)Tc ECDG uptake were higher than those of (18)F FDG uptake. Scintigraphic results demonstrated the feasibility of (99m)Tc ECDG imaging. CONCLUSION: There are similarities between (99m)Tc ECDG uptake and (18)F FDG uptake in tumors, and study findings supported the potential use of (99m)Tc ECDG as a functional imaging agent. PMID- 12563143 TI - Coronary calcification compared in patients with acute versus in those with chronic coronary events by using dual-sector spiral CT. AB - PURPOSE: To compare underlying calcific atherosclerotic lesions in acute versus chronic coronary events in patients with hypertension by using dual-sector spiral computed tomography (CT). MATERIALS AND METHODS: Eight hundred eighty-four calcific lesions were analyzed in a cohort of 50 patients (39 men, 11 women; age range, 55-79 years; mean age, 66 years +/- 6 [SD]) with hypertension who sustained a coronary event during 3-year follow-up. All underwent dual-sector spiral CT within 12 months before the event. Twenty-nine patients had an acute event (acute group): acute myocardial infarction, 20; unstable angina pectoris, six; acute ischemia, two; sudden death, one. Twenty-one patients had chronic manifestations of obstructive coronary disease (chronic group): severe stable angina, five; angiographically identified disease, 12; disease requiring angioplasty, two; and disease requiring bypass surgery, two. To examine differences between the two study groups, the chi(2) or Fisher exact test was applied to categorical parameters and the two-sample t test or Wilcoxon rank sum test to quantitative parameters. RESULTS: High prevalence of coronary calcium (total coronary calcium score [TCS] >0) was observed in both groups: 93% (27 of 29) in the acute and 95% (20 of 21) in the chronic group. There were 518 lesions in the chronic and 366 in the acute group, with a median number of 35 and nine lesions per patient, respectively (P <.001). The median TCS was 906 for the chronic and 63 for the acute group (P <.01). CONCLUSION: A mild degree of calcification characterizes patients with acute coronary events, while diffuse high-attenuation calcific plaques are associated with chronic coronary events. PMID- 12563144 TI - Small pulmonary nodules: detection at chest CT and outcome. AB - PURPOSE: To determine the outcome of pulmonary nodules less than 1 cm in diameter detected at chest computed tomography (CT). MATERIALS AND METHODS: Reports of chest CT performed during 6 months were reviewed to find patients with pulmonary nodules smaller than 1 cm in long axis for which repeat CT was recommended. Records were studied to determine whether follow-up had been performed, the initial nodules had changed in size, or nodules had been resected. RESULTS: A total of 3,446 chest CT examinations were performed, with 334 patients meeting inclusion criteria. Three patients underwent nodule resection and had pathologic examination results positive for cancer; 185 underwent follow-up, of whom 13 had results excluded as indeterminate. In the remaining 172 patients, 88 had incomplete characterization because of follow-up of less than 2 years, which left 84 with nodule characterization at follow-up. When these 84 patients were combined with the three patients with nodule resection, the number yielded was 87 patients. Seventy-seven of 87 had benign nodules because of resolution or 2-year stability, and 10 of 87 had malignant nodules because of growth or positive histologic examination results. Nine of 10 with malignant nodules had a known primary neoplasm. CONCLUSION: CT commonly helped identify small nodules. Increase in size occurred infrequently and almost exclusively in patients with a known malignancy. PMID- 12563145 TI - Analysis of 172 subtle findings on prior normal mammograms in women with breast cancer detected at follow-up screening. AB - PURPOSE: To retrospectively review nonspecific findings on prior screening mammograms to determine what features were most often deemed normal or benign despite the development of breast cancer in the same location detected at follow up screening. MATERIALS AND METHODS: Four hundred ninety-three pairs of consecutive mammographic findings were collected from 13 institutions, consisting of initial normal screening findings and a subsequent finding of cancer at screening (mean interval between examinations, 14.6 months). One designated radiologist reviewed each pair of mammograms and determined that 286 findings were judged visible at prior examination in locations where cancer later developed. Five blinded radiologists independently reviewed the prior findings in these 286 cases, identifying 169 mammograms (172 cancers) with findings so subtle that none or only one or two of the five radiologists recommended screening recall. Two unblinded radiologists reviewed the initial and subsequent findings and recorded descriptors and assessments for each finding and subjective factors influencing why, although the lesion was perceptible, it might have been undetected or not recalled. RESULTS: Of 172 cancers, 129 (75%) were invasive (112 T1 tumors and 17 T2 tumors or higher; median diameter, 10 mm), and 43 (25%) were ductal carcinoma in situ (median size, 10 mm). On the prior mammograms, 80% (137 of 172) of these cancers had subtle nonspecific findings where cancer later developed, and most were assessed as being normal or benign in appearance. CONCLUSION: There is a subset of cancers that display perceptible but nonspecific mammographic findings that do not warrant recall, as judged by both a majority of blinded radiologists and by unblinded reviewers. We believe failure to act on these nonspecific findings prospectively does not necessarily constitute interpretation below a reasonable standard of care. PMID- 12563146 TI - Breast lesions on sonograms: computer-aided diagnosis with nearly setting independent features and artificial neural networks. AB - PURPOSE: To develop a computer-aided diagnosis (CAD) algorithm with setting independent features and artificial neural networks to differentiate benign from malignant breast lesions. MATERIALS AND METHODS: Two sets of breast sonograms were evaluated. The first set contained 160 lesions and was stored directly on the magnetic optic disks from the ultrasonographic (US) system. Four different boundaries were delineated by four persons for each lesion in the first set. The second set comprised 111 lesions that were extracted from the hard-copy images. Seven morphologic features were used, five of which were newly developed. A multilayer feed-forward neural network was used as the classifier. Reliability, extendability, and robustness of the proposed CAD algorithm were evaluated. Results with the proposed algorithm were compared with those with two previous CAD algorithms. All performance comparisons were based on paired-samples t tests. RESULTS: The area under the receiver operating characteristic curve (A(z)) was 0.952 +/- 0.014 for the first set, 0.982 +/- 0.004 for the first set as the training set and the second set as the prediction set, 0.954 +/- 0.016 for the second set as the training set and the first set as the prediction set, and 0.950 +/- 0.005 for all 271 lesions. At the 5% significance level, the performance of the proposed CAD algorithm was shown to be extendible from one set of US images to the other set and robust for both small and large sample sizes. Moreover, the proposed CAD algorithm was shown to outperform the two previous CAD algorithms in terms of the A(z) value. CONCLUSION: The proposed CAD algorithm could effectively and reliably differentiate benign and malignant lesions. The proposed morphologic features were nearly setting independent and could tolerate reasonable variation in boundary delineation. PMID- 12563148 TI - Case 55: fetus in fetu. PMID- 12563149 TI - Acute appendicitis: effect of increased use of CT on selecting patients earlier. AB - PURPOSE: To explore the possibility of a relationship between increased use of computed tomography (CT) for diagnosis of appendicitis and increased occurrence of minimal or subtle CT and surgical findings. MATERIALS AND METHODS: Two groups, each with 50 consecutive patients who underwent CT before appendectomy in 1997 and 2000, were compared. CT scans and surgery-pathology reports were evaluated on a six-grade scale from normal to abscess or inflammatory mass. The demographics, surgical techniques, hospital stay, and grade distribution for the two groups were compared, and the CT results were correlated with surgical findings. RESULTS: In 1997, CT was performed in 33% (50 of 152) of patients undergoing appendectomy, as compared with 59% (50 of 85) 3 years later (P <.001). There was excellent correlation between surgical-pathologic and CT grades (weighted kappa, 0.75; P <.001; Spearman rank correlation, 0.83). There was no significant difference in demographics, rate of surgery, or surgical techniques used, but there was a significant decrease in the median surgical-pathologic grades, from 3.0 to 2.5 (P =.05) for all patients and from 3.5 to 2.6 (P =.003) for patients who underwent CT. Similarly, the median CT grade decreased from 4 to 3 (P <.001). Seven patients had subtle CT findings in 1997 compared with 16 in 2000 (P =.004), and there was a significant decrease in mean hospital stay, from 2.8 days +/- 4 (SD) to 1.5 days +/- 2 (P =.008). CONCLUSION: With increased CT use, there were less severe imaging findings, including absence of periappendiceal stranding, and a significant decrease in surgical-pathologic severity of appendiceal disease and hospital stay. PMID- 12563150 TI - Malignant gastrointestinal stromal tumor: distribution, imaging features, and pattern of metastatic spread. AB - PURPOSE: To investigate and describe the anatomic distribution, imaging features, and pattern of metastatic spread of malignant gastrointestinal stromal tumors (GISTs). MATERIALS AND METHODS: The medical records of all patients at our institution with a histologic diagnosis of GIST were reviewed. Two radiologists with knowledge of the diagnosis reviewed the radiologic findings by means of consensus. Sixty-seven patients underwent computed tomography, and scans of the primary tumor were available in 38 patients. RESULTS: One hundred sixteen patients with malignant GISTs were identified (76 men and 40 women; mean age, 54.6 years +/- 13.5 [SD]). The primary tumor locations in descending order of frequency were the small bowel (n = 49), stomach (n = 43), colon (n = 7), rectum (n = 6), other (n = 3), and not specified (n = 8). Mean primary tumor size was 13 cm +/- 6. Tumors were typically well defined (31 of 36 [86%]), with a heterogeneous rim of soft tissue with lower signal intensity than that of the contrast material-enhanced liver. Central fluid attenuation was seen in 24 of 36 (67%) patients. Metastases were seen in 23 of 38 (61%) patients at presentation and in 53 of 61 (87%) patients during follow-up. Spread was usually to the liver or peritoneum. Visceral obstruction rarely occurred, even in the presence of extensive peritoneal metastatic disease. Ascites was an unusual finding. CONCLUSION: Malignant GISTs are typically large, well-circumscribed, heterogeneous, centrally necrotic tumors that arise in the wall of the small bowel or stomach. They rarely obstruct viscera, despite their large size and propensity to metastasize to the liver and peritoneum. PMID- 12563151 TI - Detection of primary hepatic malignancy in liver transplant candidates: prospective comparison of CT, MR imaging, US, and PET. AB - PURPOSE: To determine and compare the diagnostic performance of computed tomography (CT), magnetic resonance (MR) imaging, ultrasonography (US), and positron emission tomography (PET) in the detection of hepatocellular carcinoma (HCC) or cholangiocarcinoma in liver transplant candidates and to determine interobserver variability between the readers. MATERIALS AND METHODS: Twenty-five patients were examined prospectively with CT, MR imaging, US, and PET. Each test result was interpreted independently by two radiologists. Explanted liver specimens were examined histologically to determine presence and type of lesion. Results were analyzed on a patient-by-patient basis with marginal homogeneity and effect likelihood ratio tests. RESULTS: HCC was diagnosed in nine patients. US diagnostic performance was superior to that of CT and MR imaging on a patient-by patient basis. Sensitivities were higher for US (0.89 for both US readers) than they were for CT (0.67 and 0.56 for readers 1 and 2, respectively), MR imaging (0.56 and 0.50 for readers 1 and 2, respectively), and PET (0 for both readers). None of the differences (within test) between readers were significant (P >or=.32). Ratings by US and MR observers and one CT observer were significantly associated with truth (P .05). Limb salvage rate in women exceeded that in men (93% versus 88%) at 10 years. Subgroup analysis by conduit also revealed no difference in patency. CONCLUSION: Infrainguinal arterial reconstruction can be performed safely with comparable results in women and men. Although women may present older and more often for limb salvage, outcomes do not appear to be adversely affected. PMID- 12563214 TI - The influence of peripheral vascular disease on the carotid and femoral wall mechanics in subjects with abdominal aortic aneurysm. AB - AIM: Aortic and carotid stiffness is elevated in patients with abdominal aortic aneurysm (AAA). Peripheral vascular disease (PVD) frequently coexists with AAA and may further impair the arterial wall mechanics and increase the cardiovascular load. We therefore studied the elastic carotid and muscular femoral biomechanical properties and intima-media thickness (IMT) in this group of patients. METHODS: The elastic indices and IMTs of the common carotid and common femoral arteries were determined in 30 patients with AAA (15 with PVD) with a duplex scanner coupled with a wall tracking system. Fasting plasma creatinine level, glucose and lipid concentrations, and their physiologic variables known to influence the arterial wall mechanics were also assessed. RESULTS: Patients with AAA and PVD have significantly stiffer carotid (Petersen's elastic modulus, 2207 +/- 905 mm Hg versus 1268 +/- 432 mm Hg; P =.001; stiffness index, 22.73 +/- 9.63 versus 12.60 +/- 4.24; P =.001] and femoral (Petersen's elastic modulus, 4906 +/- 4057 mm Hg versus 2599 +/- 1169 mm Hg; P =.043; stiffness index, 49.02 +/- 40.04 versus 26.07 +/- 13.22; P =.044) arteries than subjects with AAA alone. Although patients with PVD have thicker carotid and femoral IMTs, no statistical difference was seen between the two groups. The subjects were matched for age, body mass index, heart rate, systolic and diastolic blood pressures, total vascular risk score, plasma creatinine level, and fasting lipid and glucose concentrations. CONCLUSION: Subjects with PVD and AAA have significantly stiffer carotid and femoral arteries, which may indicate increased cardiovascular load and may account for the highest mortality rate seen in these patients in the UK Small Aneurysm Trial. Therefore, treatment of associated cardiovascular risk factors is important and may have to be tailored on an individual basis according to the findings of the arterial wall mechanics. PMID- 12563215 TI - Evaluation of outcomes in chronic venous disorders of the leg: development of a scientifically rigorous, patient-reported measure of symptoms and quality of life. AB - OBJECTIVE: The purpose of this study was to develop a practical and scientifically rigorous, patient-reported outcome measure to evaluate quality of life and symptoms across the range of conditions (eg, telangiectasias, varicose veins, edema, skin changes, leg ulcers) in chronic venous disorders of the leg (CVDL). METHODS: This study was a psychometric study within the VEnous INsufficiency Epidemiological and Economic Study (VEINES), an international, prospective cohort study to evaluate clinical outcomes, quality of life, costs, and use of health services in CVDL. The study was set in the 166 general practices and 116 specialist clinics in Belgium, France, Italy, and Canada (Quebec) that participated in the VEINES study plus in additional specialist clinics in Ottawa and Montreal. Field testing was carried out in three samples of patients in four countries (Belgium, France, Italy, Canada), including participants in the VEINES study (n dagger 1531) and patients recruited in additional samples of 88 English-speaking patients (Canada) and 53 French speaking patients (Belgium, France). The reliability and validity sample (n = 615) included 527 VEINES patients and 88 patients from the supplementary English speaking sample. The test-retest sample (n = 135) included 53 French-speaking and 82 English-speaking patients from the supplementary samples. The responsiveness sample included 1516 VEINES patients. The 26-item VEINES-QOL/Sym is a new, patient-reported questionnaire to evaluate symptoms and quality of life and is available in four language versions (English, French, Italian, French Canadian). RESULTS: Standard psychometric tests confirmed the acceptability (missing data, item endorsement frequencies, floor and ceiling effects), reliability (internal consistency, item-total, inter-item correlations) and validity (content, construct, convergent, discriminant, known groups) of the four language versions of the VEINES-QOL/Sym and the test-retest reliability of the English and French versions and provided preliminary evidence of responsiveness in a pooled language sample. CONCLUSION: The VEINES-QOL/Sym is a practical and scientifically sound, patient-reported measure of outcomes in CVDL that has been developed with rigorous methods. As the only fully validated measure of quality of life and symptoms that is appropriate for use across the full spectrum of CVDL-related conditions, that is quick and easy to administer, and that is available in four languages, the VEINES-QOL/Sym provides a rigorous tool for improving the evaluation of outcomes in clinical trials, epidemiologic studies, and audit. PMID- 12563216 TI - Effect of elastic compression stockings on venous hemodynamics during walking. AB - PURPOSE: Venous hemodynamics evaluated during walking better reflect changes that occur under active physiologic conditions than do conventional static modes of exercise such as tip-toe exercise, knee bending, or dorsiflexion. We prospectively studied the efficacy of air-plethysmography (APG) in monitoring venous hemodynamics during ambulation, and with this method we determined the hemodynamic effects of graduated elastic compression stockings on the lower limb during walking at various speeds. METHODS: The residual volume fraction (RVF%) during treadmill walking was monitored with APG in 10 limbs with primary chronic venous insufficiency (CVI)(CEAP(2-4)) at four speeds (1.0, 1.5, 2.0 and 2.5 km/h consecutively), with and without elastic compression (21 mm Hg at the ankle). The method was validated in comparison with standard APG, which is based on tip-toe exercise. RVF obtained during treadmill walking at 1.5 km/h was correlated with RVF measured with standard APG in 30 subjects: 12 healthy volunteers, 11 patients with primary CVI, and 7 postthrombotic limbs. Data were analyzed with nonparametric statistics. RESULTS: RVF measurements during walking were reproduced with an intra-day coefficient of variation of 5.1% to 16.5%. RVF during walking correlated well with RVF during standard APG (tip-toe) (r = 0.5, P =.004). At each of the investigated walking speeds, stockings improved venous hemodynamics by decreasing RVF, from a median of 50.5% without stockings to 40.5% with stockings at 1.0 km/h (19.8% decrease), from 49% to 39.5% at 1.5 km/h (19.4% decrease), from 50.5% to 41% at 2.0 km/h (18.8% decrease), and from 53% to 45.5% at 2.5 km/h (14.2% decrease) (all speeds, P <.02). Efficacy of the stockings in decreasing RVF (percent change in RVF) was similar across the spectrum of examined speeds (P =.47). During walking with elastic stockings, nominal RVF values were also similar across the spectrum of walking speeds, except at 2.5 km/h (P =.012). During walking without stockings, RVF did not change with treadmill speed, nor did it differ from that obtained with conventional APG (tip toe) (P =.46). The percentage decrease in RVF generated with elastic stockings correlated with the venous filling index (r = 0.73, P =.017) at 1.0 km/h. CONCLUSIONS: APG is a reproducible and valid method for monitoring venous hemodynamics during walking. Graduated elastic compression stockings significantly improved venous hemodynamics by reducing RVF in limbs with primary CVI at all examined walking speeds (1.0 to 2.5 km/h). The effect was linearly correlated with the amount of reflux (1.0 km/h). The modified application of APG during walking offers a new noninvasive method for assessment of venous hemodynamics in limbs with CVI, enabling quantification of the actual effect of elastic compression therapy during ambulation. PMID- 12563217 TI - Does ischemic preconditioning reduce spinal cord injury because of descending thoracic aortic occlusion? AB - OBJECTIVE: Ischemic preconditioning has been found to protect various organs from a subsequent longer ischemic insult. We investigated whether the late phase of ischemic preconditioning reduces spinal cord injury from occlusion of the descending thoracic aorta. METHODS: Twenty-four pigs (27 to 30 kg) were randomly divided in four groups: group I (n = 4) underwent a sham operation, group II (n = 4) underwent aortic occlusion for 20 minutes, group III (n = 8) underwent aortic occlusion for 35 minutes, and group IV (n = 8) underwent aortic occlusion for 20 minutes and, 48 hours later, aortic occlusion for 35 minutes. Aortic occlusion was accomplished with two balloon occlusion catheters placed fluoroscopically at T(6) to T(8) above the diaphragm and at the aortic bifurcation. Neurologic evaluation was performed by an independent observer according to Tarlov's scale (0 to 4, with 4 as normal). The lower thoracic and lumbar spinal cords were harvested at 120 hours and examined histologically with hematoxylin and eosin stain. Histologic results (number of neurons and grade of inflammation) were scored 0 to 4 (4, intact spinal cord; 0, no neurons and high inflammation) and were similarly analyzed. Results were expressed as the mean +/- the standard error of the mean, and statistical analysis used the Kruskal-Wallis test. RESULTS: Group IV had a better neurologic outcome at 24, 48, and 120 hours in comparison with group III (P <.001), although 120 hours after the end of the experiment, the neurologic outcome in group IV was worse than at 24 hours (P =.014). The histologic changes were proportional to the neurologic test scores, with the more severe and extensive gray matter damage in the animals of group III (number of neurons, P <.001; and grade of inflammation, P <.001). CONCLUSION: Ischemic preconditioning (late phase, 48 hours after the first occlusion) reduces spinal cord injury after aortic occlusion, as estimated with Tarlov's score and histopathology. PMID- 12563218 TI - Overexpression of von Willebrand factor is an independent risk factor for pathogenesis of intimal hyperplasia: preliminary studies. AB - OBJECTIVE: Deposition of von Willebrand factor (vWF) is increased in hyperplastic intima of grafts, vWF levels are elevated in patients with cardiovascular diseases, and there is resistance to progression of atherosclerosis in pigs with von Willebrand disease. We hypothesize that increased expression of endothelial vWF has mitogenic effects on smooth muscle cell (SMC) proliferation. METHODS: In an in vitro study, mouse aortic smooth muscle cells (SMC) were exposed to vWF in various concentrations (0, 5, 20, 100, 500, and 1000 ng/mL). DNA synthesis of SMC was measured with (3)H-thymidine incorporation. In an in vivo study, 108 mice from inbred strains of C57BL/6J (control) and RIIIS/J (characteristic of low plasma vWF) underwent carotid artery ligation (flow cessation model) and were divided into three groups: C57BL/6J, RIIIS/J, and RIIIS/J treated with desmopressin (DDAVP; intraperitoneal injection at 3 micro g/kg/d). At 2 and 4 weeks, carotid arteries were harvested for analysis with immunohistochemical analysis, morphometric studies, and reverse transcriptase polymerase chain reaction; plasma vWF was measured with an enzyme-linked immunosorbent assay. RESULTS: In vitro SMC proliferation showed a positive dose-response curve with vWF stimulation. Intimal hyperplasia (IH) in carotid arteries was prominent in C57BL/6J mice, absent in RIIIS/J mice, and moderate in RIIIS/J treated with DDAVP (intima-media ratio, 71% +/- 18%, 0, and 32% +/- 12%, respectively; P <.01). vWF deposition occurred in all hyperplastic intima subjacent to intact endothelium. Plasma vWF correlated with degree of IH (110% +/- 10%, 21% +/- 7%, and 45% +/- 8%, respectively; P <.05). vWF-messenger RNA was 9 times higher in carotid arteries of C57BL/6J mice and 4 times higher in RIIIS/J with DDAVP, compared with RIIIS/J. CONCLUSIONS: vWF directly stimulates SMC proliferation in vitro via a direct dose-response effect. In vivo low shear stress accelerates IH proportional to vWF expression. This could occur under intact endothelium without platelet activation and platelet-derived growth factor release. In effect, control of IH may entail modulation of vWF expression. PMID- 12563219 TI - The cleaved peptide of PAR1 is a more potent stimulant of platelet-endothelial cell adhesion than is thrombin. AB - PURPOSE: Platelet-endothelial cell adhesion is an important pathologic response to vessel injury or inflammation. On binding to its endothelial or platelet G protein-linked seven-transmembrane domain receptor, protease-activated receptor-1 (PAR1), thrombin releases a 41-amino acid peptide (TR(1-41)). We examined the effect of TR(1-41) on platelet activation and on platelet-endothelial cell adhesion. METHODS: A monolayer of confluent human saphenous vein endothelial cells was incubated with washed human platelets. Platelets were stimulated with either TR(1-41), TR(21-41), scrambled TR(1-41), adenosine diphosphate (ADP) epinephrine (EPI), thrombin, or thrombin receptor activating peptide (TRAP). Platelet activation was identified with flow cytometry. The magnitude of platelet endothelial cell adhesion was determined with a laser scanning cytometer that scanned the monolayer of endothelial cells and identified fluorescently bound platelets. RESULTS: Maximal thrombin stimulation (0.1 U/mL) induced a threefold increase in platelets bound to endothelial cells compared with buffer alone. Stimulation with TR(1-41) (20 mmol/L) tripled the number of platelets bound to endothelial cells compared with thrombin. Scrambled sequence of TR(1-41) (20 mmol/L) and TR(21-41) (20 mmol/L), neither of which induces platelet activation, had minimal effect on platelet adhesion. Both TRAP (20 mmol/L) and ADP-EPI (20 mmol/L) induced less platelet-endothelial cell adhesion than did thrombin. TR(1 41)-induced platelet-endothelial cell adhesion was partially blocked by glycoprotein (GP)IIb-IIIa-specific monoclonal antibody, 10E5 (10 mg/mL). CONCLUSIONS: TR(1-41), the cleaved peptide of PAR1, is a more potent stimulant of platelet-endothelial cell adhesion than is thrombin, TRAP, or ADP-EPI, and this adhesion is at least in part mediated by the platelet GPIIb-IIIa receptor. PMID- 12563220 TI - Oxidative damage and reduction of redox factor-1 expression after transient spinal cord ischemia in rabbits. AB - OBJECTIVE: The mechanism of spinal cord injury has been thought to be related to the vulnerability of spinal motor neuron cells against ischemia. However, the mechanisms of such vulnerability are not fully understood. We previously reported that spinal motor neurons may be lost by programmed cell death and thus now investigate a possible mechanism of neuronal death with immunohistochemical analysis for 8-hydroxy-2'-deoxyguanosine (8-OHdG) and redox factor-1 (Ref-1). METHODS: We used a rabbit spinal cord ischemia model with a balloon catheter. The spinal cord was removed at 8 hours, 1, 2, or 7 days after 15 minutes of transient ischemia, and histologic changes were studied with hematoxylin-eosin staining. Western blot analysis for Ref-1, temporal profiles of 8-OHdG and Ref-1 immunoreactivity, and double-label fluorescence immunocytochemical studies were performed. RESULTS: Most motor neurons were preserved until 2 days but were selectively lost at 7 days of reperfusion. Western blot analysis of a sample from sham control spinal cord showed a characteristic 37-kDa band that was reduced after ischemia. Immunohistochemistry showed the nuclear expression of Ref-1 in motor neurons of control spinal cords, and immunoreactivity was decreased 1 day after ischemia. On the other hand, no nuclear expression was seen of 8-OHdG in motor neurons of control spinal cords, and immunoreactivity was increased 1 day after ischemia. Double-label fluorescence immunocytochemical study revealed that both 8-OHdG and Ref-1 were positive at 8 hours of reperfusion in the same motor neurons, which eventually die. CONCLUSION: These results suggest that Ref-1 decreased in motor neurons after transient spinal cord ischemia and that this reduction preceded oxidative DNA damage. The reduction of Ref-1 protein at the moderately late stage of reperfusion may be one of the factors responsible for the delay in neuronal death after spinal cord ischemia. PMID- 12563221 TI - Preoperative erythropoietin and blood conservation management for thoracoabdominal aneurysm repair in a Jehovah's Witness. PMID- 12563222 TI - Treatment of a primary type IA endoleak with a liquid embolic system under conditions of aortic occlusion. AB - We present the case of a primary type IA endoleak after deployment of a bifurcated Ancure endograft (Guidant Endovascular Solutions, Menlo Park, Calif) to treat a 9-cm abdominal aortic aneurysm with a short angulated neck. The endoleak was treated unsuccessfully with repeat balloon angioplasty, placement of a Palmaz aortic stent (Cordis Endovascular, Miami, Fla), and deployment of an AneuRx aortic extender cuff (Medtronic AneuRx, Santa Rosa, Calif). The endoleak then was sealed with injection of n-butyl cyanoacrylate into the aneurysm sac at the site of the leak with occlusion of aortic flow. We suggest the use of this liquid embolic agent be considered as an adjunct to control primary type IA endoleaks when other forms of therapy have failed. PMID- 12563223 TI - Aortoduodenal fistula occurring after type II endoleak treatment with coil embolization of the aortic sac. AB - A bifurcated stent graft device was successfully deployed to exclude an asymptomatic abdominal aortic aneurysm (AAA) with adequate proximal aortic neck morphology. At 6 months, a type II endoleak was successfully embolized through a proximal perigraft channel with metallic coils. The patient was seen with upper gastrointestinal bleeding and a pulsatile abdominal mass 11 months later. Surgical exploration revealed an aortoduodenal fistula in the vicinity of the previous embolization. We discuss the possible causes of this complication and review the literature on the subject. We conclude that aortoduodenal fistula can occur after endovascular AAA repair despite the absence of endoleak or AAA diameter increase on follow-up computed tomographic scan. PMID- 12563224 TI - Aortoduodenal fistula and associated rupture of abdominal aortic aneurysm after endoluminal stent graft repair. AB - The main complications of endovascular repair of abdominal aortic aneurysms are vascular leaks and rupture, although infection and aortoduodenal fistulas have also been reported rarely. We report a case of aortoduodenal fistula with separate retroperitoneal rupture of an abdominal aortic aneurysm after endovascular stent graft repair. The initial implantation was uneventful, without any leaks at 1 month. The patient underwent open repair and did well. To our knowledge, this is the first case report of aortoduodenal fistula and associated retroperitoneal rupture of the aneurysm after endovascular stent graft repair of an abdominal aortic aneurysm. PMID- 12563225 TI - Peroneal-plantar artery bypass: a prone approach. AB - The potential space behind the distal tibia and ventral to the Achilles' tendon provides a route for bypass grafts to the plantar artery. Grafts may follow the communicating branch of the distal peroneal artery to reach the medial aspect of the foot. A prone approach for distal bypasses from the peroneal artery may facilitate exposure, conserve scarce venous conduit, and avoid scarred or infected sites. This peroneal-plantar bypass route has not been described previously. This technique was successfully used in a patient with diabetes mellitus, recurrent foot ischemia, infection, and limited venous conduit after a failed femorotibial bypass. PMID- 12563226 TI - Biomaterials in the development and future of vascular grafts. AB - Recent developments in the field of tissue engineering have re-invigorated the quest for more suitable biomaterials that are applicable to novel cardiovascular devices, including small-diameter vascular grafts. This review covers both commercially available and relevant newly developed experimental materials, including elastic polymers (polyurethane), the biodegradable and bioresorbable materials, and the naturally occurring materials, focusing on their potential applications in the development of future vascular substitutes. PMID- 12563227 TI - Branched device to preserve hypogastric arterial flow with thoracoabdominal aneurysm repair. PMID- 12563228 TI - The ethics of sham surgery in research. PMID- 12563229 TI - Regarding "impaired vasoreactivity despite an increase in plasma nitrite in patients with abdominal aortic aneurysm". PMID- 12563231 TI - Decision analysis models: opening the black box. PMID- 12563232 TI - Determining an appropriate threshold for referral to surgery for gastroesophageal reflux disease. AB - BACKGROUND: Persistent symptomatic gastroesophageal reflux disease (GERD) can be treated with medication or surgery. The purposes of this study were (1) to determine how poor the quality of life on medication would need to be to justify assuming the risks of surgery, and (2) to estimate the proportion of patients currently on medication whose quality of life is below this value. METHODS: We developed a Markov decision analysis model to simulate health outcomes (measured in quality adjusted life years [QALY]) over 10 years for medication and surgery in patients with typical GERD symptoms. We included probabilities of events obtained from a systematic literature review. Quality of life adjustments, expressed as utilities, were drawn from a survey of 131 patients 1 to 5 years after antireflux surgery. By using this model, we calculated what quality of life on medications would change the optimal strategy from medication to surgery (threshold). To determine the proportion of patients below this value, we prospectively surveyed 40 medically treated GERD patients at our hospital. RESULTS: Surgery resulted in more QALYs than medical therapy when the utility with medication use was below 0.90. Sensitivity analysis showed this value to be relatively insensitive to reasonable variations in surgical risks (mortality, failures, reoperation) and quality of life after surgery. Among those surveyed on medications, 48% fell below this threshold and would be predicted to benefit from surgery. CONCLUSION: Our model suggests that surgery would likely benefit a high proportion of medically treated GERD patients. Individual assessment of quality of life with GERD should be considered to aid clinical decision making. PMID- 12563233 TI - Residency program models, implications, and evaluation: results of a think tank consortium on resident work hours. PMID- 12563234 TI - Treatment response and prognosis of patients after recurrence of esophageal cancer. AB - BACKGROUND: Although radical operation and adjuvant chemoradiotherapy improve survival in patients with advanced esophageal cancer, more than half of these patients have recurrence. The aim of this study was to explore treatment responses and prognostic factors in patients with recurrent esophageal cancer. METHODS: The operative specimens from 258 patients undergoing radical esophagectomy with extended lymphadenectomy for esophageal squamous cell carcinoma between 1990 and 1999 were analyzed. Depth of tumor invasion, and the extent and location of lymph node metastases were determined. Postoperative recurrence was identified from positive findings on successive 3-month examinations of tumor markers, 6-month examinations of ultrasonography, and annual computed tomography. Of 258 patients, 95 had recurrence by the end of 2000 (mean follow-up was 22 months, range, 2-113). Of those 95 patients, 76 received nonsurgical treatment, 7 received operative intervention, and 12 received no treatment. Clinicopathologic features of recurrent tumors were analyzed to determine prognostic values. Serum anti-p53 antibodies (S-p53-Abs), serum C reactive protein concentration (S-CRP), and albumin concentration were also analyzed. RESULTS: The main recurrent patterns were nodal (n = 45) and organ (n = 35). Of the nonsurgical treatment group, 47 patients received chemoradiotherapy; 17, chemotherapy; and 12, radiotherapy. Overall clinical response was observed in 26 of 76 patients (34%). Treatment response was significantly associated with the type of recurrence, history of perioperative adjuvant therapy, time of recurrence, number of recurrent tumors, albumin concentration, S-CRP, and S-p53 Abs. Multivariate analysis suggested that S-p53-Abs and S-CRP were independent prognostic factors. CONCLUSION: The status of S-p53-Abs and S-CRP may predict response and outcome of patients with recurrence of esophageal cancer after radical operation. PMID- 12563235 TI - Clinical outcomes and fiscal consequences of bilateral neck exploration for primary idiopathic hyperparathyroidism without preoperative radionuclide imaging or minimally invasive techniques. AB - BACKGROUND: Radionuclide imaging-directed, minimally invasive parathyroid operation is promoted in the surgical literature and public domain as the fastest, most successful, and cheapest means of treating primary idiopathic hyperparathyroidism. The validity of these claims is unproven. This study reviews the treatment outcome results of a large series of patients treated with standard parathyroid operation without preoperative localization studies. Cost comparisons are made between this series and previous reports of selected patients in whom preoperative radionuclide imaging preceded minimally invasive parathyroid operation. METHODS: Diagnosis, treatment, and outcome data for 688 consecutive patients undergoing first neck exploration for primary idiopathic hyperparathyroidism were prospectively collected. All patients in our series underwent standard bilateral neck exploration without preoperative localization studies. Intraoperative methylene blue was used to aid identification of all parathyroid glands. Surgical findings, pathological diagnosis, operative time, length-of-stay, and treatment success data were collected. Cost data were calculated for our series using the identical calculations used in previous reports. Our outcome and calculated cost data were compared with previous reports by centers advocating scan-directed, minimally invasive parathyroid operation. RESULTS: Of 2,752 predicted total glands, 2,520 (91.6%) were identified using standard neck exploration without radionuclide localization studies. Single adenoma, with at least 1 normal gland, was found in 542 patients (78.8%), with 8 in a fifth gland. Multiple-gland hyperplasia was identified in 98 patients (14.2%) and of these 22 (3.2%) were double adenomas. Ten patients had parathyroid carcinoma (1.5%), and all received definitive surgical treatment during the primary operation. Cure rates were assessed by measurement of normal serum calcium and parathyroid hormone levels at 3 and 12 months after operation, and were 97.7% in our series. Mean operating time for the entire series was 65 minutes, decreased to 35 minutes in patients with single adenomas, and mean recovery room time was 30 minutes. Mean total costs for patients undergoing standard exploration for single adenoma was US dollars 1,107, and increased to US dollars 1,243 when patients with multigland disease, hyperplasia, or malignancy were included. CONCLUSIONS: Our series demonstrates operative times and treatment outcomes with costs that are approximately one-third less than those for scan directed, minimally invasive operation for primary idiopathic hyperparathyroidism. Thus, claims that scan-directed parathyroid operation is the cheapest, fastest, and most successful means of treatment are not supported by these data. PMID- 12563236 TI - Technical complications of continuous intra-arterial chemotherapy with 5 fluorodeoxyuridine and 5-fluorouracil for colorectal liver metastases. AB - BACKGROUND: Intra-arterial chemotherapy is an effective modality to treat unresectable hepatic metastases from colorectal primaries if systemic chemotherapy has failed. Response rates of more than 40% and a median survival of 15 to 25 months have been reported from randomized trials. In this retrospective study, we analyzed specific technical complications associated with continuous intra-arterial chemotherapy for colorectal liver metastases. METHODS: From 1982 to 1995, single-center clinical data from 180 patients with colorectal liver metastases were evaluated. Continuous intra-arterial chemotherapy was administered using either an implanted infusion pump or an intra-arterial port with an external infusion pump. The intra-arterial catheter was implanted according to the Watkins' technique. The treatment protocols consisted of 5 fluorouracil- or 5-fluorodeoxyuridine-based regimens. RESULTS: A total of 70 patients (39%) received an intra-arterial infusion pump and 110 patients (61%) an intra-arterial port. Sixty-eight technical complications affected port systems (62%), whereas 29 patients with pumps (41%) were affected by technical complications. Therapy-relevant complications were observed in 47% of the ports and 30% of the infusion pumps. The median complication-free survival was 12.2 months for infusion pumps and 7.3 months for ports (P =.0016). CONCLUSIONS: Our data demonstrate that pumps are superior to ports in terms of complication rate and complication-free survival. On the basis of our results, pumps have a potential for a longer treatment, which may result in a prolonged median survival. PMID- 12563237 TI - Supplemental perioperative fluid administration increases tissue oxygen pressure. AB - BACKGROUND: Wound infections are common and serious surgical complications. Wound perfusion delivers oxygen, inflammatory cells, growth factors, and cytokines to injured tissues. Hypoperfused regions experience low oxygen tensions that do not support adequate oxidative killing or wound healing. Clinicians may fail to recognize clinically important hypovolemia because hemodynamic stability and urine output are maintained after peripheral perfusion is compromised. We tested the hypothesis that supplemental fluid administration during and after elective colon resection increases tissue perfusion and tissue oxygen pressure. METHODS: Fifty-six patients undergoing colon resection were randomly assigned to conservative (8 mL x kg(-1) x h(-1), n = 26) or aggressive (16 to 18 mL x kg(-1) x h(-1), n = 30) fluid management. Anesthetic technique was standardized. We used 60% nitrous oxide in 40% oxygen. During surgery and postanesthetic recovery, subcutaneous oxygen tension (P(sq)O(2)) was measured by using a polarographic sensor implanted subcutaneously into 1 upper arm. Capillary blood flow was evaluated postoperatively with a thermal diffusion system. Data were analyzed with 2-tailed t tests; P value less than.05 was considered statistically significant. RESULTS: Hemodynamic and renal responses were similar in the groups. Intraoperative tissue oxygen tension was significantly greater in patients given supplemental fluid: 81 +/- 26 vs 67 +/- 18 mm Hg, P =.03. Postoperative P(sq)O(2) (77 +/- 26 vs 59 +/- 15 mm Hg, P =.009) and capillary blood flow (69 +/- 12 vs 53 +/- 12, P <.001) were also greater in the supplemental fluid patients. CONCLUSIONS: Supplemental perioperative fluid administration significantly increases tissue perfusion and tissue oxygen partial pressure. Optimizing tissue perfusion will require providing more fluid than indicated by normal clinical criteria or use of invasive monitoring to guide treatment. The actual effect of supplemental fluid administration on incidence of wound infection requires further investigation. PMID- 12563238 TI - A prospective evaluation of anorectal function after total mesorectal excision in patients with a rectal carcinoma. AB - BACKGROUND: Rectum resection with total mesorectal excision (TME) and neorectal anastomosis often compromises anorectal function. Insight into the underlying mechanisms is lacking. Therefore, a prospective study was designed to investigate the relationship between clinical and functional outcomes preoperatively and postoperatively. METHODS: Eleven patients with rectal cancer were examined before and 4 and 12 months after surgery and compared with 11 healthy volunteers (HVs). Anorectal (neorectal) function was examined by clinical outcome questionnaire, anal manometry, rectal compliance, and sensation. Six HVs also underwent barostat measurements in the sigmoid colon. RESULTS: Clinical parameters of soiling and passive incontinence (loss of stool without sensation) increased significantly until 12 months postoperatively, whereas urgency and tenesmus increased temporarily, returning to preoperative values at 12 months. In anorectal measurements, anal sphincter function was grossly preserved; however, rectal-anal inhibitory reflex (RAIR) was decreased at 4 months but recovered after 1 year. Neorectal compliance was similar to that of HV sigmoid, increasing slightly after 12 months but still significantly lower than that of normal rectum. Neorectal sensation to pressure distention was similar to that of normal rectum, however accompanied by smaller volumes. Neorectal distention induced contractions of large amplitude at 4 months, returning to normal after 12 months. CONCLUSIONS: Our results suggest that the transient increase in urgency and tenesmus after surgery results from a temporary increase in neorectal "irritability" accompanied by some adaptation of compliance in time. In contrast, episodes of incontinence and soiling are increased after 1 year most likely because of reduced neorectal capacity and RAIR recovery in the presence of a low basal anal sphincter pressure. PMID- 12563239 TI - Editorial comment: total mesorectal excision for all rectal cancers? PMID- 12563240 TI - Editorial comment: feasibility of central gastrectomy for gastric cancer. AB - BACKGROUND: Central gastrectomy (CG) for gastric cancer was developed to preserve pyloric function and maintain a large gastric volume. Whether this procedure is feasible for limited cases of gastric cancer is unclear. METHODS: On the basis of Union Internationale Contre le Cancer TNM classification, pathologic characteristics, perioperative parameters, and long-term results, we analyzed 100 patients who underwent CG. RESULTS: Pathologic findings included T1 (tumor depth, mucosal or submucosal) in 82 patients and T2 (muscularis propria or subserosal) in 18 patients. Mean number of dissected lymph nodes was 17.3, and pathologic N1 (node metastasis, 6 or less) was found in 14 patients. There were no operative deaths, but 5 patients had postoperative complications: anastomotic leakage in 1, severe gastric stasis in 2, ischemic gastric ulcer in 1, and intra-abdominal bleeding in 1. No patient had a cancer recurrence in a mean follow-up of 49 months. New early gastric cancer was detected in 3 patients during follow-up endoscopic examination. The 5-year cumulative survival was 0.97. One year after CG, 63 patients had early satiety after food intake. Mean ratio of 1-year postoperative/preoperative body weight was 95%. CONCLUSIONS: Central gastrectomy with sufficient node dissection resulted in good long-term survival and minimal postoperative weight loss. CG is a safe and useful procedure for selected patients with gastric cancer, although close follow-up for recurrence and a more precise analysis on physiologic states is needed. PMID- 12563241 TI - Mammaglobin-A is a tumor-associated antigen in human breast carcinoma. AB - BACKGROUND: Mammaglobin-A is an attractive target for immune-based therapy for patients with breast cancer because of its exclusive expression in breast cancer. In this study, we attempted to identify immunogenic T cell epitopes restricted by human leukocyte antigen (HLA)-A2 in mammaglobin-A protein. METHODS: To identify HLA-A2-restricted immunogenic epitopes from mammaglobin-A, 7 candidate peptides were synthesized and tested for immunogenicity. Each peptide was tested for binding to HLA-A2 in a HLA-A2 stabilization assay. Furthermore, T lymphocytes from 7 healthy donors and 1 patient with breast cancer received 3 weekly stimulations with autologous peptide-pulsed dendritic cells. Stimulated T cells were tested for specific recognition of peptide and tumor cells by interferon gamma enzyme-linked immunosorbent assay. RESULTS: HLA-A2 binding assays showed that all designed peptides could bind to HLA-A2. Two of the 7 peptides (MAM3 and MAM7) successfully induced peptide-specific T cells. However, only MAM3-specific T cells recognized the mammaglobin overexpressing breast cancer cell line, MDA415 transfected with HLA-A2. In contrast, MAM3-specific T cell did not recognize wild type MDA415 or MDA415 transfected with HLA-A24, or the mammaglobin negative, HLA A2 positive breast cancer cell line, MCF-7. CONCLUSIONS: Mammaglobin-A-derived peptide, MAM3, can induce mammaglobin-A-specific immunity and could be useful for vaccine strategies for patients with breast cancer. PMID- 12563242 TI - Is ischemic preconditioning of the kidney clinically relevant? AB - BACKGROUND: Renal ischemic preconditioning (IPC) is a phenomenon whereby a brief period of ischemia and reperfusion (I/R) provides tolerance to subsequent periods of ischemia. IPC has been demonstrated to protect rodent kidneys during I/R. The applicability to large mammals, including human beings, is unclear. The objective of this study was to determine if renal IPC has a beneficial effect in a large animal model of warm I/R and hypothermic preservation injury, which occurs with renal allografting. METHODS: Renal ischemia (45 minutes) and reperfusion was studied in untreated dogs and in dogs receiving IPC (10-minute/10-minute I/R). IPC was administered immediately before I/R (early IPC) or 24 hours before I/R (delayed IPC). In another group of dogs, pharmacologically induced IPC was attempted with local intra-arterial administration of dipyridamole (2.4 mg/kg/min) to increase local adenosine concentrations. Finally, IPC was induced in kidneys before harvest, cold stored for 24 hours in University of Wisconsin flush solution, and subsequently reperfused for 4 hours in allogeneic recipients. Renal functional parameters, including vascular resistance, glomerular filtration rate, urine production, oxygen consumption, and proximal tubular fluid reabsorption, were monitored during the reperfusion period and were compared with the control ischemic group. RESULTS: Renal function significantly declined during I/R, relative to the nonischemic contralateral kidney but was not different with any form of IPC, relative to the ischemic control group not treated with IPC. IPC pretreatment also did not affect the preservation injury observed in cold-stored kidneys reperfused after transplantation. CONCLUSIONS: It is concluded that IPC has no significantly measurable effects in warm or hypothermic renal I/R injury in large animals. The clinical usefulness of IPC in human renal ischemic conditions remains uncertain. PMID- 12563243 TI - Ischemia/reperfusion injury of skeletal muscle: plasma taurine as a measure of tissue damage. AB - BACKGROUND: Cell membrane rupture by oxygen-derived free radicals is a systematic feature of ischemia/reperfusion (I/R) injury. High taurine concentration gradients in skeletal muscle prompted us to evaluate whether plasma taurine levels (pTau) are a useful marker of I/R injury after different periods of ischemia. METHODS: Rabbits were randomly assigned to either 1 or 2.5 hours of hind-limb ischemia followed by 2 hours of reperfusion (groups IR1 [n = 12] and IR2.5 [n = 13], respectively). Corresponding sham groups (SHAM1 [n = 8] and SHAM2.5 [n = 9]) were used as controls. Analyzed parameters included histomorphometry and electron microscopy of skeletal muscle biopsies, pTau, and plasma level of malondialdehyde. Skeletal muscle function was assessed 3 weeks after I/R injury. RESULTS: No significant morphologic changes were detectable at the end of ischemia. After reperfusion, mild interstitial edema with intact muscle cell membranes developed in IR1 group; pTau was not increased. IR2.5 group, by contrast, showed severe interstitial edema formation (interfiber area increased by 112%, P <.005), microvascular constriction (microvessel area decreased by 33%, P <.0005), and damage to the muscle cell membranes that was confirmed by the increased plasma malondialdehyde. pTau was higher than in the SHAM2.5 group (P <.0005). Pronounced cell damage in IR2.5 group resulted in impaired muscle function (maximal tetanic tension was reduced 2 times, P <.005) but not in IR1 group. CONCLUSION: Skeletal muscle tolerates 1 h/2 h but not 2.5 h/2 h of I/R, the latter resulting in interstitial edema formation, microvascular constriction, and a late muscle dysfunction. Cell membrane rupture through stimulated lipid peroxidation promotes leakage of intracellular taurine, leading to increased pTau after reperfusion and may be considered as prognostically unfavorable in terms of organ function reversibility. In the rabbit model, pTau seems to be a sensitive marker of I/R injury to skeletal muscle. PMID- 12563245 TI - A surgeon's obligations to a Jehovah's Witness child. PMID- 12563244 TI - Tumor necrosis factor-alpha and interleukin-6 reduce bile canalicular contractions of rat hepatocytes. AB - BACKGROUND: Surgeons sometimes encounter hyperbilirubinemia without mechanical obstruction of the biliary tree postoperatively. Many of these patients have bacterial infections and endotoxemia. Kupffer's cells stimulated by endotoxin secrete inflammatory cytokines such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-6. We hypothesized that TNF-alpha and IL-6 might be involved in the pathogenesis of hyperbilirubinemia. METHODS: Effects of TNF-alpha and IL-6 on the contractions of bile canaliculi (BC) of rat hepatocyte couplets were examined and time-lapse images using phase-contrast microscopy were taken. Bile was collected from rats treated with or without the cytokines. The livers, perfused with lanthanum after the injection of cytokines, were examined ultrastructurally using electron microscopy. RESULTS: The number of BC contractions decreased in the couplets treated with both cytokines. The rapid movement of a droplet from BC was observed at the intercellular space of the hepatocyte couplet treated with TNF-alpha. Systolic blood pressure and hepatic tissue blood flow of rats injected with TNF-alpha were not changed, whereas the hepatic tissue blood flow of rats treated with IL-6 decreased (Dunnett test, P <.05). Bile secretion was reduced in both groups of rats (Dunnett test, P <.05). In rats treated with TNF-alpha the total serum bile acid concentration increased and lanthanum temporarily accumulated in BC. CONCLUSIONS: These results suggest that TNF-alpha and IL-6 may reduce BC contractions and thereby decrease bile flow. PMID- 12563246 TI - Francis Daniels Moore. PMID- 12563247 TI - Hemosuccus pancreaticus. PMID- 12563248 TI - Inverted Meckel's diverticulum. PMID- 12563249 TI - Successful steroid therapy for postoperative mesenteric panniculitis. PMID- 12563250 TI - Rapunzel syndrome causing gastric emphysema and small bowel obstruction. PMID- 12563251 TI - Sedation for patient in persistent vegetative state. PMID- 12563255 TI - Women in surgery. PMID- 12563253 TI - Ethical dilemmas in surgery series. PMID- 12563256 TI - Overinterpretation of the role of cytokeratin 19 RT-PCR of sentinel nodes in breast carcinoma. PMID- 12563257 TI - Lineage relationship and protective immunity of memory CD8 T cell subsets. AB - Memory CD8 T cells can be divided into two subsets, central (T(CM)) and effector (T(EM)), but their lineage relationships and their ability to persist and confer protective immunity are not well understood. Our results show that T(CM) have a greater capacity than T(EM) to persist in vivo and are more efficient in mediating protective immunity because of their increased proliferative potential. We also demonstrate that, following antigen clearance, T(EM) convert to T(CM) and that the duration of this differentiation is programmed within the first week after immunization. We propose that T(CM) and T(EM) do not necessarily represent distinct subsets, but are part of a continuum in a linear naive --> effector --> T(EM) --> T(CM) differentiation pathway. PMID- 12563258 TI - PI3K and Btk differentially regulate B cell antigen receptor-mediated signal transduction. AB - Phosphoinositide-3 kinase (PI3K) is thought to activate the tyrosine kinase Btk. However, through analysis of PI3K-/- and Btk-/- mice, B cell antigen receptor (BCR)-induced activation of Btk in mouse B cells was found to be unaffected by PI3K inhibitors or by a lack of PI3K. Consistent with this observation, PI3K-/- Btk-/- double-deficient mice had more severe defects than either single-mutant mouse. NF-kappaB activation along with Bcl-xL and cyclin D2 induction were severely blocked in both PI3K-/- and Btk-/- single-deficient B cells. Transgenic expression of Bcl-xL restored the development and BCR-induced proliferation of B cells in PI3K-/- mice. Our results indicate that PI3K and Btk have unique roles in proximal BCR signaling and that they have a common target further downstream in the activation of NF-kappaB. PMID- 12563259 TI - CDR3 loop flexibility contributes to the degeneracy of TCR recognition. AB - T cell receptor (TCR) binding degeneracy lies at the heart of several physiological and pathological phenomena, yet its structural basis is poorly understood. We determined the crystal structure of a complex involving the BM3.3 TCR and an octapeptide (VSV8) bound to the H-2K(b) major histocompatibility complex molecule at a 2.7 A resolution, and compared it with the BM3.3 TCR bound to the H-2K(b) molecule loaded with a peptide that has no primary sequence identity with VSV8. Comparison of these structures showed that the BM3.3 TCR complementarity-determining region (CDR) 3alpha could undergo rearrangements to adapt to structurally different peptide residues. Therefore, CDR3 loop flexibility helps explain TCR binding cross-reactivity. PMID- 12563260 TI - Pten inactivation alters peripheral B lymphocyte fate and reconstitutes CD19 function. AB - Phosphoinositide 3-kinase (PI3K) and phosphatase and tensin homolog (PTEN) phosphatase serve essential functions in the regulation of cell growth, differentiation and survival by modulating intracellular phosphatidylinositol 3,4,5-trisphosphate (PI-3,4,5-P3) concentrations. Here we show that the conditional deletion of Pten in B cells led to the preferential generation of marginal zone (MZ) B cells and B1 cells. PTEN-deficient B cells were hyperproliferative in response to mitogenic stimuli, and exhibited a lower threshold for activation through the B cell antigen receptor. Inactivation of PTEN rescued germinal center, MZ B and B1 cell formation in CD19-/- mice, arguing that recruitment and activation of PI3K are the dominant roles for CD19 in these B cell subpopulations. These findings establish the central role of PI-3,4,5-P3 regulation in the differentiation of peripheral B cell subsets. PMID- 12563261 TI - Essential role of Src-family protein tyrosine kinases in NF-kappaB activation during B cell development. AB - The nature of signals that govern the development of immunoglobulin heavy chain dependent B cells is largely unknown. Using mice deficient for the B cell expressed Src-family protein tyrosine kinases (SFKs) Blk, Fyn and Lyn, we show an essential role of these kinases in pre-B cell receptor (pre-BCR)- mediated NF kappaB activation and B cell development. This signaling defect is SFK specific, as a deficiency in Syk, which controls pre-B cell development, does not affect NF kappaB induction. Impaired NF-kappaB induction was overcome by the activation of protein kinase C (PKC)-lambda, thus suggesting the involvement of PKC-lambda in pre-BCR-mediated SFK-dependent activation of NF-kappaB. Our data show the existence of a functionally distinct SFK signaling module responsible for pre-BCR mediated NF-kappaB activation and B cell development. PMID- 12563262 TI - A role for CK2 in the Drosophila circadian oscillator. AB - The posttranslational modification of clock proteins is critical for the function of circadian oscillators. By genetic analysis of a Drosophila melanogaster circadian clock mutant known as Andante, which has abnormally long circadian periods, we show that casein kinase 2 (CK2) has a role in determining period length. Andante is a mutation of the gene encoding the beta subunit of CK2 and is predicted to perturb CK2beta subunit dimerization. It is associated with reduced beta subunit levels, indicative of a defect in alpha:beta association and production of the tetrameric alpha2:beta2 holoenzyme. Consistent with a direct action on the clock mechanism, we show that CK2beta is localized within clock neurons and that the clock proteins Period (Per) and Timeless (Tim) accumulate to abnormally high levels in the Andante mutant. Furthermore, the nuclear translocation of Per and Tim is delayed in Andante, and this defect accounts for the long-period phenotype of the mutant. These results suggest a function for CK2 dependent phosphorylation in the molecular oscillator. PMID- 12563263 TI - Identification and function of thermosensory neurons in Drosophila larvae. AB - Although the ability to sense temperature is critical for many organisms, the underlying mechanisms are poorly understood. Using the calcium reporter yellow cameleon 2.1 and electrophysiological recordings, we identified thermosensitive neurons and examined their physiologic response in Drosophila melanogaster larvae. In the head, terminal sensory organ neurons showed increased activity in response to cooling by < or =1 degrees C, heating reduced their basal activity, and different units showed distinct response patterns. Neither cooling nor heating affected dorsal organ neurons. Body wall neurons showed a variety of distinct response patterns to both heating and cooling; the diverse thermal responses were strikingly similar to those described in mammals. These data establish a functional map of thermoresponsive neurons in Drosophila larvae and provide a foundation for understanding mechanisms of thermoreception in both insects and mammals. PMID- 12563264 TI - Combinations of muscle synergies in the construction of a natural motor behavior. AB - A central issue in motor control is how the central nervous system generates the muscle activity patterns necessary to achieve a variety of behavioral goals. The many degrees of freedom of the musculoskeletal apparatus provide great flexibility but make the control problem extremely complex. Muscle synergies- coherent activations, in space or time, of a group of muscles--have been proposed as building blocks that could simplify the construction of motor behaviors. To evaluate this hypothesis, we developed a new method to extract invariant spatiotemporal components from the simultaneous recordings of the activity of many muscles. We used this technique to analyze the muscle patterns of intact and unrestrained frogs during kicking, a natural defensive behavior. Here we show that combinations of three time-varying muscle synergies underlie the variety of muscle patterns required to kick in different directions, that the recruitment of these synergies is related to movement kinematics, and that there are similarities among the synergies extracted from different behaviors. PMID- 12563266 TI - Cloning terror. PMID- 12563265 TI - Translocation of CaM kinase II to synaptic sites in vivo. PMID- 12563267 TI - Signalling pathways that mediate skeletal muscle hypertrophy and atrophy. AB - Atrophy of skeletal muscle is a serious consequence of numerous diseases, including cancer and AIDS. Successful treatments for skeletal muscle atrophy could either block protein degradation pathways activated during atrophy or stimulate protein synthesis pathways induced during skeletal muscle hypertrophy. This perspective will focus on the signalling pathways that control skeletal muscle atrophy and hypertrophy, including the recently identified ubiquitin ligases muscle RING finger 1 (MuRF1) and muscle atrophy F-box (MAFbx), as a basis to develop targets for pharmacologic intervention in muscle disease. PMID- 12563268 TI - Killer wiles: growing interest in Fas. PMID- 12563269 TI - How to attract a sperm. PMID- 12563270 TI - Caveat experimentor--is your myosin really inhibited? PMID- 12563271 TI - ATMachine. PMID- 12563272 TI - Mitochondria, AIF and caspases--rivaling for cell death execution. PMID- 12563275 TI - Developmental control of cell morphogenesis: a focus on membrane growth. AB - To date, the role of transport and insertion of membrane in the control of membrane remodelling during cell and tissue morphogenesis has received little attention. In contrast, the contributions of cytoskeletal rearrangements and both intercellular and cell-substrate attachments have been the focus of many studies. Here, we review work from many developmental systems that highlights the importance of polarized membrane growth and suggests a general model for the role of endocytic recycling during cell morphogenesis. We also address how the spatio temporal control of membrane insertion during development can account for various classes of tissue rearrangements. We suggest that tubulogenesis, tissue spreading and cell intercalation stem mostly from a remarkably small number of cell intrinsic surface remodelling events that confer on cells different modes of migratory behaviours. PMID- 12563276 TI - The signal flow and motor response controling chemotaxis of sea urchin sperm. AB - The signalling pathway and the behavioural strategy underlying chemotaxis of sperm are poorly understood. We have studied the cellular events and motor responses that mediate chemotaxis of sperm from the sea urchin Arbacia punctulata. Here we show that resact, a chemoattractant peptide, initiates a rapid and transient rise in the concentration of cyclic GMP, followed by a transient influx of Ca2+. The binding of a single resact molecule elicits a Ca2+ response, and 50-100 bound molecules saturate the response. The ability to register single molecules is reminiscent of the single-photon sensitivity of rod photoreceptors. Both resact and cyclic nucleotides cause a turn or brief tumbling in the swimming path of sperm. We conclude that a cGMP-mediated increase in the Ca2+ concentration induces the primary motor response of sperm to the chemoattractant. PMID- 12563277 TI - Native Myosin-IXb is a plus-, not a minus-end-directed motor. AB - Myosin-IXb (Myo9b) is a single-headed, processive motor that contains a Rho GTPase-activating protein (GAP) domain within its tail. Although tail-less myosin IXb motor domain moves towards the minus end of the actin filament, we show here that full-length myosin-IXb is a plus-end-directed motor. This suggests that the tail domain of myosin-IXb regulates motor directionality. PMID- 12563278 TI - In situ assembly of enzyme inhibitors using extended tethering. AB - Cysteine aspartyl protease-3 (caspase-3) is a mediator of apoptosis and a therapeutic target for a wide range of diseases. Using a dynamic combinatorial technology, 'extended tethering', we identified unique nonpeptidic inhibitors for this enzyme. Extended tethering allowed the identification of ligands that bind to discrete regions of caspase-3 and also helped direct the assembly of these ligands into small-molecule inhibitors. We first designed a small-molecule 'extender' that irreversibly alkylates the cysteine residue of caspase-3 and also contains a thiol group. The modified protein was then screened against a library of disulfide-containing small-molecule fragments. Mass-spectrometry was used to identify ligands that bind noncovalently to the protein and that also form a disulfide linkage with the extender. Linking the selected fragments with binding elements from the extenders generates reversible, tight-binding molecules that are druglike and distinct from known inhibitors. One molecule derived from this approach inhibited apoptosis in cells. PMID- 12563279 TI - Site-specific cassette exchange and germline transmission with mouse ES cells expressing phiC31 integrase. AB - Currently two site-specific recombinases are available for engineering the mouse genome: Cre from P1 phage and Flp from yeast. Both enzymes catalyze recombination between two 34-base pair recognition sites, lox and FRT, respectively, resulting in excision, inversion, or translocation of DNA sequences depending upon the location and the orientation of the recognition sites. Furthermore, strategies have been designed to achieve site-specific insertion or cassette exchange. The problem with both recombinase systems is that when they insert a circular DNA into the genome (trans event), two cis-positioned recognition sites are created, which are immediate substrates for excision. To stabilize the trans event, functional mutant recognition sites had to be identified. None of the systems, however, allowed efficient selection-free identification of insertion or cassette exchange. Recently, an integrase from Streptomyces phage phiC31 has been shown to function in Schizosaccharomyces pombe and mammalian cells. This enzyme recombines between two heterotypic sites: attB and attP. The product sites of the recombination event (attL and attR) are not substrates for the integrase. Therefore, the phiC31 integrase is ideal to facilitate site-specific insertions into the mammalian genome. PMID- 12563281 TI - The impact of systemic infection on the progression of neurodegenerative disease. PMID- 12563280 TI - Binding, spatial attention and perceptual awareness. PMID- 12563283 TI - The role of histamine and the tuberomamillary nucleus in the nervous system. PMID- 12563282 TI - Wnts and TGF beta in synaptogenesis: old friends signalling at new places. PMID- 12563284 TI - Neurobiology of butyrylcholinesterase. PMID- 12563285 TI - Human prefrontal cortex: processing and representational perspectives. PMID- 12563286 TI - Neuroethics: an agenda for neuroscience and society. PMID- 12563287 TI - NALPs: a novel protein family involved in inflammation. AB - A newly discovered family of cytoplasmic proteins--the NALPs--has been implicated in the activation of caspase-1 by the Toll-like receptors (TLRs) during the cell's response to microbial infection. Like the structurally related apoptotic protease-activating factor-1 (APAF-1), which is responsible for the activation of caspase-9, the NALP1 protein forms a large, signal-induced multiprotein complex, the inflammasome, resulting in the activation of pro-inflammatory caspases. PMID- 12563288 TI - Acquisition of siderophores in gram-negative bacteria. AB - The outer membrane of Gram-negative bacteria constitutes a permeability barrier that protects the cell from exterior hazards, but also complicates the uptake of nutrients. In the case of iron, the challenge is even greater, because of the scarcity of this indispensable element in the cell's surroundings. To solve this dilemma, bacteria have evolved sophisticated mechanisms whereby the concerted actions of receptor, transporter and energy-transducing proteins ensure that there is a sufficient supply of iron-containing compounds, such as siderophores. PMID- 12563289 TI - Tor signalling in bugs, brain and brawn. AB - TOR--a highly conserved atypical protein kinase and the 'target of rapamycin', an immunosuppressant and anti-cancer drug--controls cell growth. TOR controls the growth of proliferating yeast, fly and mammalian cells in response to nutrients. Recent findings, however, indicate that TOR also controls the growth of non proliferating cells, such as neurons and muscle cells. Furthermore, TOR, by associating with regulatory proteins and inhibiting phosphatases, controls the activity of multiphosphorylated effectors. PMID- 12563290 TI - Temporal and spatial coordination of exocytosis and endocytosis. AB - In secretory cells, exocytosis and compensatory endocytosis are tightly coupled membrane trafficking processes that control the surface area and composition of the plasma membrane. While exocytic and endocytic processes have been studied independently in great detail, at present there is much interest in understanding the mode of their coupling. This review discusses emerging insights into the coupling of these processes, both in the chemical synapses of neurons and in non neuronal cells. PMID- 12563291 TI - Transglutaminases: crosslinking enzymes with pleiotropic functions. AB - Blood coagulation, skin-barrier formation, hardening of the fertilization envelope, extracellular-matrix assembly and other important biological processes are dependent on the rapid generation of covalent crosslinks between proteins. These reactions--which are catalysed by transglutaminases--endow the resulting supramolecular structure with extra rigidity and resistance against proteolytic degradation. Some transglutaminases function as molecular switches in cytoskeletal scaffolding and modulate protein-protein interactions. Having knowledge of these enzymes is essential for understanding the aetiologies of diverse hereditary diseases of the blood and skin, and various autoimmune, inflammatory and degenerative conditions. PMID- 12563292 TI - Bioinformatics goes back to the future. AB - The need to turn raw data into knowledge has led the bioinformatics field to focus increasingly on the manipulation of information. By drawing parallels with both cryptography and artificial intelligence, we can develop an understanding of the changes that are occurring in bioinformatics, and how these changes are likely to influence the bioinformatics job market. PMID- 12563293 TI - Caveolae--from ultrastructure to molecular mechanisms. AB - Almost 50 years after the first sighting of small pits that covered the surface of mammalian cells, investigators are now getting to grips with the detailed workings of these enigmatic structures that we now know as caveolae. PMID- 12563294 TI - Hide, shield and strike back: how HIV-infected cells avoid immune eradication. AB - Viruses that induce chronic infections can evade immune responses. HIV is a prototype of this class of pathogen. Not only does it mutate rapidly and make its surface components difficult to access by neutralizing antibodies, but it also creates cellular hideouts, establishes proviral latency, removes cell-surface receptors and destroys immune effectors to escape eradication. A better understanding of these strategies might lead to new approaches in the fight against AIDS. PMID- 12563295 TI - Alloreactive killer cells: hindrance and help for haematopoietic transplants. AB - Haematopoietic-cell transplantation is a treatment for leukaemia and lymphoma. To reduce the incidence of graft-versus-host disease (GVHD) caused by transplanted T cells, donors and recipients are HLA matched. For patients for whom a matched donor is not available, one option is transplantation from an HLA-mismatched relative who shares one HLA haplotype. This procedure is distinguished by the use of a stronger conditioning regimen for the patient and of a T-cell-depleted graft containing numerous stem cells. After transplantation, natural killer cells are prevalent, and they can include alloreactive cells that kill tumour cells and prevent GVHD. The alloreactions seem to be determined by the mismatched HLA class I ligands and their killer-cell immunoglobulin-like receptors. PMID- 12563296 TI - CD3-specific antibody-induced active tolerance: from bench to bedside. AB - Although they were used initially as non-specific immunosuppressants in transplantation, CD3-specific monoclonal antibodies have elicited renewed interest owing to their capacity to induce immune tolerance. In mouse models of autoimmune diabetes, CD3-specific antibodies induce stable disease remission by restoring tolerance to pancreatic beta-cells. This phenomenon was extended recently to the clinic--preservation of beta-cell function in recently diagnosed patients with diabetes was achieved by short-term administration of a CD3 specific antibody. CD3-specific antibodies arrest ongoing disease by rapidly clearing pathogenic T cells from the target. Subsequently, they promote long-term T-cell-mediated active tolerance. Recent data indicate that transforming growth factor-beta-dependent CD4+CD25+ regulatory T cells might have a central role in this effect. PMID- 12563297 TI - Interleukin-12 and the regulation of innate resistance and adaptive immunity. AB - Interleukin-12 (IL-12) is a heterodimeric pro-inflammatory cytokine that induces the production of interferon-gamma (IFN-gamma), favours the differentiation of T helper 1 (T(H)1) cells and forms a link between innate resistance and adaptive immunity. Dendritic cells (DCs) and phagocytes produce IL-12 in response to pathogens during infection. Production of IL-12 is dependent on differential mechanisms of regulation of expression of the genes encoding IL-12, patterns of Toll-like receptor (TLR) expression and cross-regulation between the different DC subsets, involving cytokines such as IL-10 and type I IFN. Recent data, however, argue against an absolute requirement for IL-12 for T(H)1 responses. Our understanding of the relative roles of IL-12 and other factors in T(H)1-type maturation of both CD4+ and CD8+ T cells is discussed here, including the participation in this process of IL-23 and IL-27, two recently discovered members of the new family of heterodimeric cytokines. PMID- 12563298 TI - The complementary roles of deletion and regulation in transplantation tolerance. AB - Neonatal tolerance of alloantigens was described in mice nearly half a century ago, but unfortunately, the translation of these early findings into the clinical arena proved to be much more challenging than was first anticipated. However, the past decade has seen considerable progress in our understanding of the mechanisms that contribute to transplantation tolerance in experimental models. This review outlines our current understanding of the mechanisms of allograft tolerance, emphasizing the complementary roles of deletion and regulation of alloreactive T cells. PMID- 12563299 TI - Semaphorins in interactions between T cells and antigen-presenting cells. AB - Although semaphorins were identified originally as guidance cues for developing neuronal axons, accumulating evidence indicates that several semaphorins are expressed also in the immune system. SEMA4D (CD100), which is expressed constitutively by T cells, enhances the activation of B cells and dendritic cells (DCs) through its cell-surface receptor, CD72. SEMA4A, which is expressed by DCs, is involved in the activation of T cells through interactions with TIM2. So, these semaphorins seem to function in the reciprocal stimulation of T cells and antigen-presenting cells (APCs). Emerging evidence indicates that additional semaphorins and related molecules are involved in T-cell-APC interactions also. PMID- 12563300 TI - Innate immune sensing and its roots: the story of endotoxin. AB - How does the host sense pathogens? Our present concepts grew directly from longstanding efforts to understand infectious disease: how microbes harm the host, what molecules are sensed and, ultimately, the nature of the receptors that the host uses. The discovery of the host sensors--the Toll-like receptors--was rooted in chemical, biological and genetic analyses that centred on a bacterial poison, termed endotoxin. PMID- 12563301 TI - Kinases as therapeutic targets for heart failure. PMID- 12563302 TI - Understanding biofilm resistance to antibacterial agents. PMID- 12563303 TI - Molecular imaging in drug discovery and development. PMID- 12563304 TI - A proton-pump inhibitor expedition: the case histories of omeprazole and esomeprazole. PMID- 12563305 TI - The basics of mass spectrometry in the twenty-first century. PMID- 12563306 TI - Modern biomedical research: an internally self-consistent universe with little contact with medical reality? PMID- 12563307 TI - Protein folding and disease: a view from the first Horizon Symposium. PMID- 12563308 TI - Disruption of differentiation in human cancer: AML shows the way. AB - Although much is understood about the ways in which transcription factors regulate various differentiation systems, and one of the hallmarks of many human cancers is a lack of cellular differentiation, relatively few reports have linked these two processes. Recent studies of acute myeloid leukaemia (AML), however, have indicated how disruption of transcription-factor function can disrupt normal cellular differentiation and lead to cancer. This model involves lineage-specific transcription factors, which are involved in normal haematopoietic differentiation. These factors are often targeted in AML--either by direct mutation or by interference from translocation proteins. Uncovering these underlying pathways will improve the diagnosis and treatment of AML, and provide a working model for other types of human cancer, including solid tumours. PMID- 12563309 TI - Inhibiting the p53-MDM2 interaction: an important target for cancer therapy. AB - p53 is an attractive therapeutic target in oncology because its tumour-suppressor activity can be stimulated to eradicate tumour cells. Inhibiting the p53-MDM2 interaction is a promising approach for activating p53, because this association is well characterized at the structural and biological levels. MDM2 inhibits p53 transcriptional activity, favours its nuclear export and stimulates its degradation, so inhibiting the p53-MDM2 interaction with synthetic molecules should lead to p53-mediated cell-cycle arrest or apoptosis in p53-positive stressed cells. PMID- 12563310 TI - The endothelin axis: emerging role in cancer. AB - Collectively, the endothelins and their receptors--referred to as the endothelin (ET) axis--have key physiological functions in normal tissue, acting as modulators of vasomotor tone, tissue differentiation, development, cell proliferation and hormone production. Based on new data, the ET axis also functions in the growth and progression of various tumours. Preliminary results from clinical trials, such as those with atrasentan--an ET(A)-receptor antagonist -in prostate cancer, are encouraging. The place of ET-receptor antagonists in cancer therapy for a range of malignancies merits further investigation. PMID- 12563311 TI - The role of p53 in determining sensitivity to radiotherapy. AB - Ionizing radiation (IR) has proven to be a powerful medical treatment in the fight against cancer. Rational and effective use of its killing power depends on understanding IR-mediated responses at the molecular, cellular and tissue levels. Tumour cells frequently acquire defects in the molecular regulatory mechanisms of the response to IR, which sensitizes them to radiation therapy. One of the key molecules involved in a cell's response to IR is p53. Understanding these mechanisms indicates new rational approaches to improving cancer treatment by IR. PMID- 12563312 TI - Coordinated regulation of life and death by RB. AB - Recent studies have shown that RB can inhibit apoptosis, independently of its ability to block cell proliferation. This poses the question of how cells choose to grow or to die when RB becomes inactivated. RB is phosphorylated following mitogenic stimulation, but it is degraded in response to death stimuli. Most sporadic cancers also inactivate RB by phosphorylation, rather than losing RB entirely--possibly to exploit the survival advantage conferred by RB under stress. Drawing from the different mechanisms of RB inactivation, we propose two models for ways in which cells use RB to make the choice of life versus death. PMID- 12563313 TI - An analysis of cancer research funding in the UK. AB - Until recently, a lack of comparable and reliable data on ongoing research activity has been a significant limiting factor in strategic planning for cancer research. This article describes a new initiative in the UK, which is aimed at facilitating the coordination and strategic planning of cancer research at the national level. PMID- 12563314 TI - Host defense against Pseudomonas aeruginosa requires ceramide-rich membrane rafts. AB - Pseudomonas aeruginosa infection is a serious complication in patients with cystic fibrosis and in immunocompromised individuals. Here we show that P. aeruginosa infection triggers activation of the acid sphingomyelinase and the release of ceramide in sphingolipid-rich rafts. Ceramide reorganizes these rafts into larger signaling platforms that are required to internalize P. aeruginosa, induce apoptosis and regulate the cytokine response in infected cells. Failure to generate ceramide-enriched membrane platforms in infected cells results in an unabated inflammatory response, massive release of interleukin (IL)-1 and septic death of mice. Our findings show that ceramide-enriched membrane platforms are central to the host defense against this potentially lethal pathogen. PMID- 12563315 TI - PUMA-G and HM74 are receptors for nicotinic acid and mediate its anti-lipolytic effect. AB - Nicotinic acid (niacin), a vitamin of the B complex, has been used for almost 50 years as a lipid-lowering drug. The pharmacological effect of nicotinic acid requires doses that are much higher than those provided by a normal diet. Its primary action is to decrease lipolysis in adipose tissue by inhibiting hormone sensitive triglyceride lipase. This anti-lipolytic effect of nicotinic acid involves the inhibition of cyclic adenosine monophosphate (cAMP) accumulation in adipose tissue through a G(i)-protein-mediated inhibition of adenylyl cyclase. A G-protein-coupled receptor for nicotinic acid has been proposed in adipocytes. Here, we show that the orphan G-protein-coupled receptor, 'protein upregulated in macrophages by interferon-gamma' (mouse PUMA-G, human HM74), is highly expressed in adipose tissue and is a nicotinic acid receptor. Binding of nicotinic acid to PUMA-G or HM74 results in a G(i)-mediated decrease in cAMP levels. In mice lacking PUMA-G, the nicotinic acid-induced decrease in free fatty acid (FFA) and triglyceride plasma levels was abrogated, indicating that PUMA-G mediates the anti-lipolytic and lipid-lowering effects of nicotinic acid in vivo. The identification of the nicotinic acid receptor may be useful in the development of new drugs to treat dyslipidemia. PMID- 12563317 TI - Getting a grip on genetic testing. PMID- 12563316 TI - Activated protein C blocks p53-mediated apoptosis in ischemic human brain endothelium and is neuroprotective. AB - Activated protein C (APC) is a systemic anti-coagulant and anti-inflammatory factor. It reduces organ damage in animal models of sepsis, ischemic injury and stroke and substantially reduces mortality in patients with severe sepsis. It was not known whether APC acts as a direct cell survival factor or whether its neuroprotective effect is secondary to its anti-coagulant and anti-inflammatory effects. We report that APC directly prevents apoptosis in hypoxic human brain endothelium through transcriptionally dependent inhibition of tumor suppressor protein p53, normalization of the pro-apoptotic Bax/Bcl-2 ratio and reduction of caspase-3 signaling. These mechanisms are distinct from those involving upregulation of the genes encoding the anti-apoptotic Bcl-2 homolog A1 and inhibitor of apoptosis protein-1 (IAP-1) by APC in umbilical vein endothelial cells. Cytoprotection of brain endothelium by APC in vitro required endothelial protein C receptor (EPCR) and protease-activated receptor-1 (PAR-1), as did its in vivo neuroprotective activity in a stroke model of mice with a severe deficiency of EPCR. This is consistent with work showing the direct effects of APC on cultured cells via EPCR and PAR-1 (ref. 9). Moreover, the in vivo neuroprotective effects of low-dose mouse APC seemed to be independent of its anti-coagulant activity. Thus, APC protects the brain from ischemic injury by acting directly on brain cells. PMID- 12563318 TI - Thermotaxis of mammalian sperm cells: a potential navigation mechanism in the female genital tract. PMID- 12563319 TI - Is alloantigen expression by host epithelium required for acute graft-versus-host disease? PMID- 12563321 TI - Cyclin E is a more powerful predictor of breast cancer outcome than proliferation. PMID- 12563322 TI - US scientific panels Bush-whacked. PMID- 12563323 TI - Europe dithers over stem cell patents. PMID- 12563324 TI - PLoS snaps up Cell editor. PMID- 12563325 TI - Finding a funding niche. PMID- 12563326 TI - Scientific openness faces reality check. PMID- 12563327 TI - New Stanford institute sparks cloning quarrel. PMID- 12563328 TI - American universities operate fewer industry trials. PMID- 12563329 TI - Peter Piot. PMID- 12563331 TI - Murder mysteries in type 1 diabetes. PMID- 12563332 TI - Tat's seductive side. PMID- 12563333 TI - B cell memory: sapping the T cell. PMID- 12563334 TI - Splitting vessels: keeping lymph apart from blood. PMID- 12563335 TI - eLiXiRs for restraining inflammation. PMID- 12563336 TI - Orchestrating sleep-wake functions in the brain. PMID- 12563337 TI - Stress in the sarcomere. PMID- 12563341 TI - NTP Toxicology and Carcinogenesis Studies of Glutaraldehyde (CAS NO. 111-30-8) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Glutaraldehyde is used in large volume in a variety of industries as a disinfectant, preservative, fixative and cross-linking agent, and as a chemical intermediate in the synthesis of pharmaceuticals and pesticides. Glutaraldehyde was nominated by the National Cancer Institute, the Occupational Safety and Health Administration, and the National Institute of Environmental Health Sciences for carcinogenicity studies because of potential occupational exposure. Male and female F344/N rats and B6C3F1 mice were exposed to glutaraldehyde (25% aqueous solution) (approximately 93% pure) by inhalation for 2 years. In vitro genetic toxicology studies were conducted in Salmonella typhimurium, L5178Y mouse lymphoma cells, and cultured Chinese hamster ovary cells; in vivo studies were conducted to measure sex-linked recessive lethal mutations in Drosophila melanogaster, chromosomal aberrations and micronucleated erythrocytes in mouse bone marrow, and micronucleated erythrocytes in mouse peripheral blood. The results of 13-week inhalation studies with glutaraldehyde were reported previously (NTP, 1993 -- TOX-25 ). 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to 0, 250, 500, or 750 ppb glutaraldehyde vapor by inhalation, 6 hours per day, 5 days per week, for 104 weeks. Survival of 500 and 750 ppb female rats was less than that of the chamber controls. Mean body weights of all exposed groups of male rats and 500 and 750 ppb female rats were generally less than those of the chamber controls. Some female rats exposed to 750 ppb were thin to emaciated at the time they were killed moribund. Increased incidences of nonneoplastic nasal lesions occurred primarily within the anterior section of the nose in 500 and 750 ppb rats and to a lesser extent in 250 ppb rats. The more significant lesions included hyperplasia and inflammation of the squamous and respiratory epithelia and squamous metaplasia of the respiratory epithelium. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were exposed to 0, 62.5, 125, or 250 ppb glutaraldehyde vapor by inhalation, 6 hours per day, 5 days per week, for 104 weeks. Survival of exposed mice was similar to that of the chamber controls. Mean body weights of female mice exposed to 250 ppb were generally less than those of the chamber controls throughout the study. Incidences of squamous meta-plasia of the respiratory epithelium were increased in 250 ppb males and females and 125 ppb females. Incidences of hyaline degeneration of the respiratory epithelium were increased in all exposed groups of females. The incidence of inflammation of the nose was marginally increased in 250 ppb females. GENETIC TOXICOLOGY: In genetic toxicity studies, glutaraldehyde was muta-genic with and without S9 metabolic activation in S. typhimurium strains TA100, TA102, and TA104. Glutaraldehyde was mutagenic in mouse L5178Y lymphoma cells in the absence of S9 and induced sister chromatid exchanges in cultured Chinese hamster ovary cells with and without S9. No increase in chromosomal aberrations was induced by glutaraldehyde in cultured Chinese hamster ovary cells with or without S9 at one laboratory; at another laboratory, chromosomal aberrations were induced in the absence of S9 only. Glutaraldehyde did not induce sex-linkedrecessive lethal mutations in germ cells of male D. melanogaster treated as adults by feeding or injection or treated as larvae by feeding. In vivo, glutaraldehyde induced a significant increase in chromosomal aberrations in mouse bone marrow cells 36 hours after a single intraperitoneal injection. In a subset of the 36-hour chromosomal aberrations test, there was a small increase in the number of micronucleated bone marrow polychromatic eryth-rocytes, which was judged to be equivocal. Addi-tional short-term (3-day) and subchronic (13-week) micronucleus tests in mice, using the intraperitoneal or inhalation routes, respectively, yielded negative results. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was no evidence of carcinogenic activinogenic activity of glutaraldehyde in male or female F344/N rats exposed to 250, 500, or 750 ppb. There was no evidence of carcinogenic activity in male or female B6C3F1 mice exposed to 62.5, 125, or 250 ppb. Incidences of nonneoplastic lesions of the nose were significantly increased in male and female rats and mice. Synonyms: 1,3-Diformylpropane; glutaral; glutardialdehyde; glutaric dialdehyde; 1,5-pentanedial; 1,5-pentanedione; potentiated acid glutaraldehyde Trade names: Cidex; Sonacide. PMID- 12563342 TI - NTP Studies of Magnetic Field Promotion (DMBA Initiation) in Female Sprague Dawley Rats (Whole-body Exposure/Gavage Studies). AB - Electric and magnetic fields are associated with the production, transmission, and use of electricity; thus, the potential for human exposure is high. These elec-tric and magnetic fields are predominantly of low fre-quency (60 Hz in the United States and 50 Hz in Europe) and generally of low intensity. Because some epidemiology studies and initiation/promotion studies in rats have suggested a potential for increased breast cancer rates with increasing magnetic field exposure, the ability of 50- and 60-Hz magnetic fields to pro-mote mammary gland tumors initiated by the administration of 7,12-dimethylbenz(a)anthracene (DMBA) was examined in female Sprague-Dawley rats in 13- and 26-week whole-body exposure studies. Additional animals were evaluated for changes in pineal gland and serum melatonin concentrations. FIRST 13-WEEK STUDY: Groups of 100 female Sprague Dawley rats were ad-ministered 20 mg DMBA (four weekly gavage doses of 5 mg in sesame oil) and exposed to 1 G 50-Hz, 5 G 50-Hz, or 1 G 60-Hz magnetic fields for 18.5 hours per day, 7 days per week, for 13 weeks. A group of 100 rats administered 20 mg DMBA served as DMBA controls. A group of 100 vehicle control rats was administered only sesame oil on the same schedule. Additional groups of 10 rats receiving similar treatment were evaluated for pineal gland and serum melatonin concentrations at 4, 8, or 12 weeks. All vehicle control rats survived to the end of the study. Of the animals administered 20 mg DMBA, 6 rats in the DMBA control group, 13 in the DMBA/1 G 50-Hz group, eight in the DMBA/5 G 50-Hz group, and five in the DMBA/1 G 60-Hz group died or were removed from the study prior to the final necropsy. Final mean body weights and body weight gains of the DMBA/1 G 50-Hz and DMBA/1 G 60-Hz groups and the mean body weight gain of the DMBA/5 G 50-Hz group were slightly greater than those of the DMBA control group. Clinical findings including torso masses and ulcers (on the mammary masses) were attributed to DMBA administration. The numbers of palpable mammary gland tumors, tumor sizes, and total tumor areas in DMBA/magnetic field groups were similar to those in the DMBA control group. Relative to the DMBA control group, exposure to magnetic fields did not significantly affect overall incidences of mammary gland neoplasms or nonneoplastic lesions in the DMBA/magnetic field groups. SECOND 13 WEEK STUDY: Groups of 100 female Sprague-Dawley rats were ad-ministered 8 mg DMBA (four weekly gavage doses of 2 mg in sesame oil) and exposed to 1 G 50-Hz or 5 G 50-Hz magnetic fields for 18.5 hours per day, 7 days per week, for 13 weeks. A group of 100 female rats administered 8 mg DMBA served as DMBA controls. Additional groups of 10 rats receiving similar treatment were evaluated for pineal gland and serum melatonin concentrations at 4, 8, or 12 weeks. Except for one rat in the DMBA/5 G 50-Hz group, all rats survived until the end of the study. Final mean body weights of DMBA/magnetic field groups were similar to those of the DMBA control group. Clinical findings including torso masses and ulcers were attributed to DMBA administration. The numbers of palpable mammary gland tumors, tumor sizes, and total tumor areas in DMBA/magnetic field groups were similar to those in the DMBA control group. Relative to the DMBA control group, exposure to magnetic fields did not significantly affect overall incidences of mammary gland neoplasms or nonneoplastic lesions in the DMBA/magnetic field groups. 26-WEEK STUDY: Groups of 100 female Sprague-Dawley rats were administered 10 mg DMBA (in sesame oil) by gavage followed by exposure to 1 G 50-Hz, 5 G 50-Hz, or 1 G 60-Hz magnetic fields for 18.5 hours per day, 7 days per week, for 26 weeks. A group of 100 female rats administered 10 mg DMBA served as DMBA controls. Another 100 vehicle control rats were administered only sesame oil. Additional groups of 10 rats receiving similar treatment were evaluated for pineal gland and serum melatonin concentrations at 4, 8, or 12 weeks. All rats in the vehicle control group survived until the end of the study. Twelve rats in the DMBA control group, 15 in the DMBA/1 G 50-Hz group, 9 in the DMBA/5 G 50-Hz group, and six in the DMBA/1 G 60-Hz group died or were removed during the study. The final mean body weights and body weight gains of the DMBA/1 G 50-Hz and DMBA/5 G 50-Hz groups were significantly greater than those of the DMBA control group. Clinical findings including torso masses, abscesses, and ulcers were attributed to DMBA administration. The pineal gland melatonin concentrations of DMBA/5 G 50-Hz and DMBA/1 G 60-Hz rats were significantly greater than that of the DMBA controls at week 12; however, these data were highly variable between individual animals within each group. The numbers of palpable mammary gland tumors, tumor sizes, and total tumor areas in DMBA/magnetic field groups were similar to those in the DMBA controls. The incidences of mammary gland carci-noma (including multiple) in the DMBA/1 G 60-Hz group were significantly decreased relative to the DMBA control group. CONCLUSIONS: In an initiation/promotion study in which female Sprague-Dawley rats were initiated by four weekly doses of 5 mg DMBA per rat beginning at 50 days of age and exposed to 50-Hz magnetic fields at 1 or 5 G field intensities or to 1 G 60-Hz magnetic fields for 13 weeks, there was no evidence that magnetic fields promoted the development of mammary gland neoplasms. The prevalence and multiplicity of mammary gland carcinomas in all DMBA groups limited the ability of this assay to detect a promoting effect of magnetic fields. In an initiation/promotion study in which female Sprague-Dawley rats were initiated by four weekly doses of 2 mg DMBA per rat beginning at 50 days of age and exposed to 50-Hz magnetic fields at 1 or 5 G field intensities for 13 weeks, there was no evidence that magnetic fields promoted the development of mammary gland neoplasms. In an initiation/promotion study in which female Sprague-Dawley rats were initiated by a single 10 mg DMBA dose at 50 days of age and then exposed to 50-Hz magnetic fields at 1 or 5 G field intensities or to 1 G 60-Hz magnetic fields for 26 weeks, there was no evidence that magnetic fields promoted the development of mammary gland neoplasms. PMID- 12563343 TI - NTP Toxicology and Carcinogenesis Studies of 60-HZ Magnetic Fields IN F344/N Rats and B6C3F1 Mice (Whole-body Exposure Studies). AB - Electric and magnetic fields (EMF) are associated with the production, transmission, and use of electricity; thus, the potential for human exposure is high. These electric and magnetic fields are predominantly of low frequency (60 Hz in the United States and 50 Hz in Europe) and generally of low intensity. Epidemiology studies have suggested a potential for increased breast cancer, brain cancer, and leukemia rates with increasing magnetic field exposure. Therefore, given the widespread exposure to low-intensity, 60-Hz magnetic fields in industrialized societies, standard toxicology studies and long-term carcinogenesis studies were conducted using traditional rodent models. Male and female F344/N rats and B6C3F1mice were exposed to 60-Hz magnetic fields by whole body exposure for 2 years. 2-YEAR STUDY IN RATS: Groups of 100 male and 100 female rats were exposed to 60-Hz magnetic fields at intensities of 0.02, 2, or 10 G for 18.5 hours per day, 7 days per week, for 106 weeks. Groups of 100 male and 100 female control rats were housed in the same exposure chambers without applied magnetic fields. Additional groups of 100 male and 100 female rats were intermittently exposed (1 hour on and 1 hour off) to a 10 G 60-Hz field 18.5 hours per day, 7 days per week, for 106 weeks. The highest field intensity (10 G) is approximately 5,000-fold greater than what was considered high intensity for homes in epidemiology studies in humans. Survival and Body Weights: Survival and mean body weights of exposed groups of male and female rats was similar to those of the control groups. Pathology Findings: The incidences of thyroid gland C-cell adenoma and carcinoma in 0.02 G male rats, adenoma in 2 G males, and adenoma or carcinoma (combined) in 0.02 and 2 G males were significantly greater than in the control group. The incidence of mononuclear cell leukemia in males in the 10 G intermittent group was significantly less than in the control group. 2-YEAR STUDY IN MICE: Groups of 100 male and 100 female mice were exposed to 60-Hz magnetic fields at intensities of 0.02, 2, or 10 G for 18.5 hours per day, 7 days per week, for 106 weeks. Groups of 100 male and 100 female control mice were housed in the same exposure chambers without applied magnetic fields. Additional groups of 100 male and 100 female mice were intermittently exposed (1 hour on and 1 hour off) to a 10 G 60-Hz field 18.5 hours per day, 7 days per week, for 106 weeks. Survival and Body Weights: Survival of male mice exposed to 10 G was significantly less than that of control mice after 2 years; survival of all other exposed groups of mice was similar to that of control mice. Mean body weights of exposed groups of male and female mice were similar to those of the control groups throughout the study. Pathology Findings: The incidences of alveolar/bronchiolar adenoma were significantly decreased in 0.02 and 2 G male mice and 2 G female mice relative to the control groups; the incidences of alveolar/bronchiolar adenoma or carcinoma (combined) were significantly less in males and females exposed to 2 G than in the control groups. In female mice, the incidence of malignant lymphoma in the 10 G intermittent group was significantly less than in the controls. CONCLUSIONS: Under the conditions of these 2-year whole-body exposure studies, there was equivocal evidence of carcinogenic activity of 60-Hz magnetic fields in male F344/N rats based on increased incidences of thyroid gland C-cell neoplasms in the 0.02 and 2G groups. There was no evidence of carcinogenic activity in female F344/N rats or male or female B6C3F1 mice exposed to 0.02, 2, or 10 G, or 10 G intermittent 60-Hz magnetic fields. In exposed rats and mice there were no increased incidences of neoplasms at sites for which epidemiology studies have suggested an association with magnetic fields (brain, mammary gland, leukemia). PMID- 12563344 TI - NTP Toxicology and Carcinogenesis Studies of Isobutene (CAS No. 115-11-7) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Isobutene is produced during the fractionation of refinery gases or through the catalytic cracking of methyl-t-butyl ether. Isobutene is primarily used to produce diisobutylene, trimers, butyl rubber, and other polymers. In addition, it is used in the production of isooctane, high-octane aviation gasoline, methyl-t butyl ether, and copolymer resins with butadiene and acrylonitrile. Isobutene was selected for evaluation because of the potential for human exposure due to its large production volume and the lack of adequate data on its carcinogenic potential. The toxicity and carcinogenicity of isobutene were determined in male and female F344/N rats and B6C3F1 mice exposed to isobutene (greater than 98% pure) by inhalation for 14 weeks or 2 years. The mutagenicity of isobutene was assessed in Salmonella typhimurium, and the frequency of micronuclei was determined in the peripheral blood of mice exposed by inhalation for 14 weeks. 14 WEEK STUDIES: Groups of 10 male and 10 female F344/N rats and B6C3F1 mice were exposed to isobutene at concentrations of 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm 6 hours per day, 5 days per week, for 14 weeks. Concentrations greater than 8,000 ppm isobutene were not used because of the danger of explosion. All rats and mice survived to the end of the study. The final mean body weights and body weight gains of all exposed groups were similar to those of the chamber controls. No exposure-related gross lesions were observed in male or female rats or mice at necropsy. Microscopically, minimal hypertrophy of goblet cells lining the nasopharyngeal duct in the most caudal nose section was observed in some rats in each exposed group of males and females. 2-YEAR STUDIES: Based on the lack of significant exposure-related toxicologic effects in the 14-week rat and mouse studies, 8,000 ppm was selected as the highest exposure concentration in the 2 year studies. Concentrations of 0, 500, 2,000, and 8,000 ppm were selected for rats and mice with the 500 and 2,000 ppm selection based on published metabolic elimination rates for Sprague-Dawley rats and B6C3F1 mice. Rats: Groups of 50 male and 50 female F344/N rats were exposed to isobutene at concentrations of 0, 500, 2,000, or 8,000 ppm 6 hours per day, 5 days per week, for 105 weeks. Survival of exposed males and females was similar to that of the chamber controls. Mean body weights of exposed groups were generally similar to those of the chamber controls throughout the study. 2-Hydroxyisobutyric Acid - Biomarker of Exposure 2-Hydroxyisobutyric acid (HIBA), the major urinary metabolite of isobutene, was measured in the urine of male and female rats as an indicator of isobutene exposure at 6, 12, and 18 months. The amount of HIBA excreted increased with increasing exposure concentration. However, when HIBA concentration was normalized to isobutene exposure concentration, the relative amount of HIBA excreted decreased with increasing exposure concentration, implying nonlinear kinetics. Pathology Findings: The incidence of thyroid gland follicular cell carcinoma in male rats exposed to 8,000 ppm was increased compared to the chamber control group and exceeded the historical control range. The incidences of hyaline degeneration of the olfactory epithelium were marginally increased in exposed rats; however, the severities of hyaline degeneration increased with increasing exposure concentration in males and females. Mice: Groups of 50 male and 50 female B6C3F1 mice were exposed to isobutene at concentrations of 0, 500, 2,000, or 8,000 ppm 6 hours per day, 5 days per week, for 105 weeks. Survival of exposed males and females was similar to that of the chamber controls. Mean body weights of exposed mice were generally similar to those of the chamber controls throughout the study except for female mice exposed to 2,000 or 8,000 ppm, which weighed slightly less than chamber controls from about week 52 until week 92. 2 Hydroxyisobutyric Acid - Biomarker of Exposure: HIBA was measured in the urine of male and female mice as an indicator of isobutene exposure at 6, 12t 6, 12,and 18 months. The amount of HIBA excreted increased with increasing exposure concentration. However, when HIBA concentration was normalized to isobutene exposure concentration, the relative amount of HIBA excreted decreased with increasing exposure concentration, implying nonlinear kinetics. Pathology Findings: The incidences of hyaline degeneration of the respiratory epithelium in all groups of exposed males and females were significantly greater than those in the chamber control groups. The incidences of hyaline degeneration of the olfactory epithelium in 2,000 and 8,000 ppm mice were greater than those in the chamber controls. GENETIC TOXICOLOGY: Isobutene was not mutagenic in any of four strains of S. typhimurium, with or without S9 metabolic activation, and no increase in the frequency of micronucleated erythrocytes was seen in peripheral blood of male or female mice treated with isobutene by inhalation for 14 weeks. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was some evidence of carcinogenic activity of isobutene in male F344/N rats based on an increased incidence of follicular cell carcinoma of the thyroid gland. There was no evidence of carcinogenic activity of isobutene in female F344/N rats or male or female B6C3F1 mice exposed to 500, 2,000, or 8,000 ppm. Exposure to isobutene by inhalation for 2 years resulted in increased incidences and/or severities of nasal lesions including hyaline degeneration of the olfactory epithelium in male and female rats and mice and hyaline degeneration of the respiratory epithelium in male and female mice. Synonyms: Isobutylene, 2 methylpropene, liquified petroleum gas, g-butylene. PMID- 12563345 TI - NTP Toxicology and Carcinogenesis Studies of 2,3-Dibromo-1-Propanol (CAS No. 96 13-9) in F344/N Rats and B6C3F1 Mice (Dermal Studies). AB - 2,3-Dibromo-l-propanol, a colorless liquid, has been used as a flame retardant, as an intermediate in the preparation of the flame retardant tris(2,3 dibromopropyl) phosphate, and as an intermediate in the manufacture of pesticides and pharmaceutical preparations. Toxicology and carcinogenicity studies were conducted by applying 2,3-dibromo-1-propanol (approximately 98% pure) in ethanol to the subscapular area of the skin of male and female F344/N rats and B6C3F1 mice 5 days per week for 16 days, 13 weeks, 48 to 51 weeks (male rats), 52 to 55 weeks (female rats), 36 to 39 weeks (male mice), or 39 to 42 weeks (female mice). Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, Drosophila melanogaster, mouse Iymphoma cells, and mouse bone marrow cells. 16-DAY STUDY IN RATS Groups of five male and five female rats received dermal applications of 0, 44, 88, 177, 375, or or 750 mg/kg 2,3 dibromo-1-propanol 5 days per week for 16 days. One male and one female receiving 750 mg/kg died before the end of the study. The mean body weight gains and final mean body weights of dosed rats were similar to those of the controls. There were no clinical findings or gross lesions associated with chemical application. 16 DAY STUDY IN MICE Groups of five male and five female mice received dermal applications of 0, 44, 88, 177, 375, or 750 mg/kg 2,3-dibromo-1-propanol 5 days per week for 16 days. Four males and one female receiving 750 mg/kg died before the end of the study. The mean body weight gains and final mean body weights of dosed mice were similar to those of the controls. There were no clinical findings or gross lesions associated with chemical application. 13-WEEK STUDY IN RATS Groups of 10 male and 10 female rats received dermal applications of 0, 44, 88, 177, 375, or 750 mg/kg 2,3-dibromo-1-propanol 5 days per week for 13 weeks. All rats survived until the end of the for 16 days. One male and one female receiving study. For rats in the 750 mg/kg groups, the mean 750 mg/kg died before the end of the study. The body weight gain was 11% lower than that of the controls for males and 13% lower for females. The mean liver weights and liver-weight-to-body weight ratios of males receiving 375 or 750 mg/kg and of females receiving 750 mg/kg were increased. Chemical-related lesions occurred in the kidney of male rats and in the liver of female rats. The average severity of nephropathy was slightly increased in males receiving dermal applications of 750 mg/kg, while individual cell necrosis was observed in the liver of all female rats in the 750 mg/kg group. 13-WEEK STUDY IN MICE Groups of 10 male and 10 female mice received dermal applications of 0, 44, 88, 177, 375, or 750 mg/kg 2,3-dibromo-1-propanol 5 days per week for 13 weeks. Eight male mice receiving 750 mg/kg died during the study, while all female mice survived. The final mean body weights of dosed and control mice were similar. The mean liver weights and liver weight-to-body-weight ratios of males receiving 375 or 750 mg/kg and of females receiving 750 mg/kg were increased. Chemical-related lesions occurred in the liver and lung of mice. Centrilobular hepatocellular necrosis occurred in all males in the 750 mg/kg group that died during the study, while individual cell necrosis was observed in the liver of females receiving 177, 375, or 750 mg/kg. Pleomorphism of the epithelium in pulmonary bronchioles occurred with a dose related increased incidence in males and females. Necrosis of the bronchiolar epithelium was observed in males receiving 750 mg/kg. LONG-TERM STUDY IN RATS Originally planned to last for 2 years, the chronic study in rats was terminated early because of reduced survival in the high-dose groups related to chemical induced neoplasms and because of the detection of antibodies to Iymphocytic choriomeningitis virus in sentinel mice. Groups of 50 male and 50 female rats received dermal applications of 0,188, or 375 mg/kg 2,3-dibromo-1-propanol 5 days per week for 48 to 51 weeks (males) or 52 to 55 weeks (females). Su5 weeks (females). Survival, Body Weights, and Clinical Findings The survival of 375 mg/kg male and female rats was significantly lower than that of the controls (males: 50/50, 41/50,16/50; females: 48/50, 38/50, 24/50). In the 375 mg/kg groups, the final mean body weight was 23% lower than that of the controls for males and 14% lower for females. There were no chemical related clinical findings. Pathology Findings Application of 2,3-dibromo-1-propanol to the skin produced significant dose-related increases in the incidences of neoplasms at numerous sites in male and female rats. Almost all dosed rats had malignant neoplasms; only one control male and one control female had malignant neoplasms. In male rats, the incidences of benign or malignant neoplasms of the skin, nose, Zymbal's gland, oral mucosa, esophagus, and small and large intestines were significantly increased in the low- and high-dose groups, while the incidences of neoplasms of the forestomach and liver were significantly increased only in the high-dose group. Neoplasms of the kidney, vascular neoplasms of the spleen, and mesotheliomas in males occurred with a significant positive trend. In female rats, the incidences of benign or malignant neoplasms of the nose, Zymbal's gland, oral mucosa, esophagus, large intestine, and liver were significantly increased in the low- and high-dose groups, while the incidences of neoplasms of the skin, forestomach, small intestine, mammary gland, and clitoral gland were significantly increased in the high-dose group only. Neoplasms of the kidney in females occurred with a significant positive trend. LONG-TERM STUDY IN MICE Originally planned to last for 2 years, the chronic study in mice was terminated early because of the detection of antibodies to lymphocytic choriomeningitis virus in sentinel mice. Groups of 50 male and 50 female mice received dermal applications of 0, 88, or 177 mg/kg 2,3-dibromo-1-propanol 5 days per week for 36 to 39 weeks (males) or 39 to 42 weeks (females). Survival, Body Weights, and Clinical Findings All mice (except two low-dose females) survived until study termination. Mean body weights of control and dosed mice were similar throughout the study, and there were no clinical findings attributed to 2,3-dibromo-l propanol. Pathology Findings Application of 2,3-dibromo-1-propanol to the skin produced significant dose-related increases in the incidences of neoplasms at several sites in male and female mice. Benign or malignant neoplasms were observed in 40% of the low-dose males, 66% of the high-dose males, 52% of the low-dose females, and 56% of the high-dose females. In control groups, neoplasms occurred in 6% of the males and 10% of the females. In male and female mice, the incidences of benign or malignant neoplasms of the forestomach were significantly increased in the low- and high dose groups, while the incidences of neoplasms of the skin were significantly increased only in the high-dose groups. The incidences of liver and lung neoplasms were increased in high-dose males. GENETIC TOXICOLOGY 2,3-Dibromo-l propanol was mutagenic in a variety of short-term tests, independent of exogenous metabolic activation (S9). It induced gene mutations in three strains of Salmonella typhimurium (TA98, TA100, and TA1535) and was positive in the mouse Iymphoma assay for induction of trifluorothymidine resistance in L5178Y cells. 2,3-Dibromo-l-propanol induced sister chromatid exchanges and chromosomal aberrations in cultured Chinese hamster ovary cells. In germ cells of male Drosophila melanogaster, 2,3-dibromo-1-propanol induced sex-linked recessive lethal mutations and reciprocal translocations. Results of an in vivo bone marrow micronucleus assay in male mice treated with 2,3-dibromo-1-propanol were negative. CONCLUSIONS Under the conditions of these long-term dermal studies, there was clear evidence of carcinogenic activity of 2,3-dibromo-1-propanol in male F344/N rats based on increased incidences of neoplasms of the skin, nose, oral mucosa, esophagus, forestomach, small and large intestine, Zymbal's gland, liver, kidney, tunica vaginalis, and spleen. There was clear evidence of carcinogenic activity of 2,3-dibromo-1-propanol in female F344/N rats based on increased incidences of neoplasms of the skin, nose, oral mucosa, esophagus, forestomach, small and large intestine, Zymbal's gland, liver, kidney, clitoral gland, and mammary gland. There was clear evidence of carcinogenic activity of 2,3-dibromo-1-propanol in male B6C3F1 mice based on increased incidences of neoplasms of the skin, forestomach, liver, and lung. There was clear evidence of carcinogenic activity of 2,3-dibromo-1-propanol in female B6C3F1 mice based on increased incidences of neoplasms of the skin and the forestomach. The increased incidences of alveolar/bronchiolar adenomas in female mice may have been related to chemical administration. In rats, 2,3-dibromo-1-propanol caused increased incidences of hyperkeratosis in the skin, forestomach, and esophagus, epithelial dysplasia in the nose, pleomorphism and basophilic and clear cell changes in the liver, and nuclear enlargement in the kidney. There were also chemical-related increases in the incidences of forestomach ulcers and acanthosis, angiectasis in the liver, and renal hyperplasia in male rats and epithelial dysplasia of the forestomach and bile duct hyperplasia in the liver in female rats. Chemical related increases occurred in the incidences of hyperplasia in the skin, epithelial dysplasia of the forestomach, and bronchiolar epithelial pleomorphism and hyperplasia in male and female mice and in the incidence of eosinophilic cytoplasmic change in the liver in males. Synonyms: 2,3-dibromopropanol; 2,3 dibromopropyl alcohol. PMID- 12563346 TI - Toxicology and carcinogenesis studies of sodium nitrite (CAS NO. 7632-00-0) in F344/N rats and B6C3F1 mice (drinking water studies). AB - Sodium nitrite is used as a color fixative and preservative in meats and fish. It is also used in manufacturing diazo dyes, nitroso compounds, and other organic compounds; in dyeing and printing textile fabrics and bleaching fibers; in photography; as a laboratory reagent and a corrosion inhibitor; in metal coatings for phosphatizing and detinning; and in the manufacture of rubber chemicals. Sodium nitrite also has been used in human and veterinary medicine as a vasodilator, a bronchial dilator, an intestinal relaxant, and an antidote for cyanide poisoning. Sodium nitrite was nominated by the FDA for toxicity and carcinogenesis studies based on its widespread use in foods. Male and female F344/N rats and B6C3F1 mice were exposed to sodium nitrite (99% pure) in drinking water for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, rat and mouse bone marrow, and mouse peripheral blood. 14 WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to 0, 375, 750, 1500, 3,000, or 5000 ppm sodium nitrite (equivalent to average daily doses of approximately 30, 55, 115, 200, or 310 mg sodium nitrite/kg body weight to males and 40, 80, 130, 225, or 345 mg/kg to females) in drinking water for 14 weeks. Clinical pathology study groups of 15 male and 15 female rats were exposed to the same concentrations for 70 or 71 days. One female exposed to 3000 ppm died before the end of the study. Body weights of males exposed to 3000 or 5000 ppm and females exposed to 5000 ppm were significantly less than those of the controls. Water consumption by 5000 ppm males and 3000 and 5000 ppm females was less than that by the controls at weeks 2 and 14. Clinical findings related to sodium nitrite exposure included brown discoloration in the eyes and cyanosis of the mouth, tongue, ears, and feet of males exposed to 3000 or 5000 ppm and of females exposed to 1500 ppm or greater. Reticulocyte counts were increased in males and females exposed to 3000 or 5000 ppm. The erythron was decreased on day 19 but increased by week 14 in males and females exposed to 5000 ppm. Methemoglobin concentrations were elevated in almost all exposed groups throughout the 14 week study; a no-observed-adverse-effect level was not achieved. The relative kidney and spleen weights of males and females exposed to 3000 or 5000 ppm were significantly greater than those of the controls. Sperm motility in 1500 and 5000 ppm males was significantly decreased. Increased erythropoietic activity in the bone marrow of exposed males and females was observed. The incidences of squamous cell hyperplasia of the forestomach in 5000 ppm males and females were significantly increased. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were exposed to 0, 375, 750, 1500, 3000, or 5000 ppm sodium nitrite (equivalent to average daily doses of approximately 90, 190, 345, 750, or 990 mg/kg to males and 120, 240, 445, 840, or 1230 mg/kg to females) in drinking water for 14 weeks. Body weights of males exposed to 5000 ppm were significantly less than those of the controls. Water consumption by males exposed to 1500 ppm or greater was slightly less than that by the controls at week 13. Relative spleen weights of 3000 and 5000 ppm males and absolute and relative heart, kidney, liver, and spleen weights of females exposed to 3000 or 5000 ppm were greater than those of the control groups. Sperm motility was decreased in 5000 ppm males, and the estrous cycles of 1500 and 5000 ppm females were significantly longer than in the controls. There were increased incidences of squamous cell hyperplasia of the forestomach in 5000 ppm males and females, extramedullary hematopoiesis of the spleen in 3000 and 5000 ppm males and 1500 ppm or greater females, and degeneration of the testis in 3000 and 5000 ppm males. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed to 0, 750, 1500, or 3000 ppm sodium nitrite (equivalent to average daily doses of approximately 35, 70, or 130 mg/kg to males and 40, 8d 40, 80, or 150 mg/kg to females) in drinking water for 2 years. For toxicokinetic studies of plasma nitrite and blood methemoglobin, 10 male and 10 female special study rats were exposed to the same concentrations for 12 months. Survival of exposed groups was similar to that of the controls. Mean body weights of males and females exposed to 3000 ppm were less than those of the controls throughout the study. Water consumption by males and females exposed to 3000 ppm was less than that by the controls throughout the study, and that by the other exposed groups was generally less after week 14. The incidences of hyperplasia of the forestomach epithelium in males and females exposed to 3000 ppm were significantly greater than those in the control groups. The incidence of fibroadenoma of the mam mary gland was significantly increased in females exposed to 1500 ppm, and the incidences of multiple fibroadenoma were increased in 750 ppm and 1500 ppm females; however, these neoplasms occur with a high background incidence, and no increase was seen in the 3000 ppm group. The incidences of mononuclear cell leukemia were significantly decreased in males and females exposed to 1500 or 3000 ppm. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were exposed to 0, 750, 1500, or 3000 ppm sodium nitrite (equivalent to average daily doses of approximately 60, 120, or 220 mg/kg to males and 45, 90, or 165 mg/kg to females) in drinking water for 2 years. Survival of exposed groups was similar to that of the controls; mean body weights of 3000 ppm females were less than those of the controls throughout the study. Exposed groups generally consumed less water than the control groups. The incidences of squamous cell papilloma or carcinoma (combined) in the forestomach of female mice occurred with a positive trend. The incidence of hyperplasia of the glandular stomach epithelium was significantly greater in 3000 ppm males than in the controls. GENETIC TOXICOLOGY: Sodium nitrite was mutagenic in Salmonella typhimurium strain TA100, with and without Aroclor 1254-induced hamster and rat liver S9 enzymes; no mutagenicity was observed in strain TA98. Results of acute bone marrow micronucleus tests with sodium nitrite in male rats and mice by intraperitoneal injection were negative. In addition, a peripheral blood micronucleus assay conducted with mice from the 14-week study gave negative results. CONCLUSIONS: Under the conditions of this 2 year drinking water study, there was no evidence of carcinogenic activity of sodium nitrite in male or female F344/N rats exposed to 750, 1500, or 3000 ppm. There was no evidence of carcinogenic activity of sodium nitrite in male B6C3F1 mice exposed to 750, 1500, or 3000 ppm. There was equivocal evidence of carcinogenic activity of sodium nitrite in female B6C3F1 mice based on the positive trend in the incidences of squamous cell papilloma or carcinoma (combined) of the forestomach. Exposure to sodium nitrite in drinking water resulted in increased incidences of epithelial hyperplasia in the forestomach of male and female rats and in the glandular stomach of male mice. Decreased incidences of mononuclear cell leukemia occurred in male and female rats. PMID- 12563347 TI - NTP Toxicology and Carcinogenesis Studies of EMODIN (CAS NO. 518-82-1) Feed Studies in F344/N Rats and B6C3F1 Mice. AB - Emodin is a naturally occurring anthraquinone present in the roots and bark of numerous plants of the genus Rhamnus. Extracts from the roots, bark, and/or dried leaves of buckthorn, senna, cascara, aloe, frangula, and rhubarb have been used as laxatives since ancient times and currently are widely used in the preparation of herbal laxative preparations. Anthraquinone glycosides are poorly absorbed from the gastrointestinal tract but are cleaved by gut bacteria to produce aglycones (such as emodin) that are more readily absorbed and are responsible for the purgative properties of these preparations. There is extensive exposure to emodin and other anthraquinones resulting from the use of herb-based stimulant laxatives. Reports that 1,8-dihydroxyanthraquinone, a commonly used laxative ingredient, caused tumors in the gastrointestinal tract of rats raised the possibility of an association between colorectal cancer and the use of laxatives containing anthraquinones. Because emodin is a hydroxyanthraquinone structurally similar to 1,8-dihydroxyanthraquinone, is present in herbal laxatives, and was reported to be mutagenic in bacteria, it was considered a potential carcinogen and was selected for in-depth evaluation. Male and female F344/N rats and B6C3F1 mice were exposed to emodin (at least 94% pure) in feed for 16 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, rat and mouse bone marrow cells, and mouse peripheral blood erythrocytes. 16-DAY STUDY IN RATS: Groups of five male and five female rats were fed diets containing 0, 600, 2,000, 5,500, 17,000, or 50,000 ppm emodin (equivalent to average daily doses of approximately 50, 170, 480, 1,400, or 3,700 mg emodin/kg body weight to males and 50, 160, 460, 1,250, or 2,000 mg/kg to females) for 15 (males) or 16 (females) days. Three female rats died before the end of the study. Mean body weights of males and females exposed to 5,500 ppm or greater were significantly less than those of the controls. Feed consumption by males and females receiving 17,000 or 50,000 ppm was decreased throughout the study. Macroscopic lesions were present in the kidney of rats exposed to 17,000 or 50,000 ppm. 16-DAY STUDY IN MICE: Groups of five male and five female mice were fed diets containing 0, 600, 2,000, 5,500, 17,000, or 50,000 ppm emodin (equivalent to average daily doses of approximately 120, 400, 1,200, or 3,800 mg/kg to males and 140, 530, 1,600, or 5,000 mg/kg to females; 50,000 ppm equivalents were not calculated due to high mortality) for 15 (males) or 16 (females) days. All mice exposed to 50,000 ppm died before the end of the study. Mice in the 17,000 ppm groups lost weight during the study. Feed consumption by 5,500 ppm females was greater than that by the controls throughout the study. Macroscopic lesions were present in the gallbladder and kidney of mice exposed to 17,000 ppm. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were fed diets containing 0, 312.5, 625, 1,250, 2,500, or 5,000 ppm emodin (equivalent to average daily doses of approximately 20, 40, 80, 170, or 300 mg/kg to males and females) for 14 weeks. Mean body weights of males exposed to 2,500 ppm or greater and females exposed to 1,250 ppm or greater were significantly less than those of the controls. During the first week of the study, feed consumption by males exposed to 2,500 or 5,000 ppm and females exposed to 5,000 ppm was less than that by the controls. Feed consumption by these groups was similar to that by the controls for the remainder of the study. In rats exposed to 2,500 or 5,000 ppm, there were increases in platelet counts in males and females and segmented neutrophil counts in females. Total serum protein and albumin concentrations were decreased in females exposed to 2,500 or 5,000 ppm. Relative kidney weights of rats exposed to 1,250 ppm or greater and relative lung weights of rats exposed to 625 ppm or greater were significantly increased compared to the control groups. Relative liver weights were incree increased in females exposed to 625 ppm or greater. The estrous cycle length wassignificantly increased in females exposed to 1,250 or 5,000 ppm. All male rats exposed to 1,250 ppm or greater and all exposed female rats had pigment in the renal tubules; and the severity of pigmentation generally increased with increasing exposure concentration. The incidences of hyaline droplets in the cortical epithelial cytoplasm were increased in all groups of exposed males and in females exposed to 312.5, 625, or 1,250 ppm. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were fed diets containing 0, 312.5, 625, 1,250, 2,500, or 5,000 ppm emodin (equivalent to average daily doses of approxi mately 50, 100, 190, 400, or 800 mg/kg to males and 60, 130, 240, 500, or 1,100 mg/kg to females) for 14 weeks. All mice survived to the end of the study. Mean body weights of males exposed to 2,500 or 5,000 ppm were significantly less than those of the controls. Feed consumption by exposed groups was generally similar to that by the controls. Relative kidney weights of male mice exposed to 1,250 ppm or greater, relative lung weights of males exposed to 625 ppm or greater, and relative liver weights of female mice exposed to 625 ppm or greater were increased. The incidences and severities of nephropathy were increased in males and females exposed to 1,250 ppm or greater. The incidences of renal tubule pigmentation were significantly increased in males exposed to 625 ppm or greater and in females exposed to 1,250 ppm or greater. 2-YEAR STUDY IN RATS: Groups of 65 male and 65 female rats were fed diets containing 0, 280, 830, or 2,500 ppm emodin (equivalent to average daily doses of approximately 110, 320, or 1,000 mg/kg to males and 120, 370, or 1,100 mg/kg to females) for 105 weeks. Ten male and ten female rats from each group were necropsied at 6 months. Blood samples from five male and five female rats in each group were evaluated at 3, 6, and 12 months for plasma emodin concentrations; these rats were necropsied at 12 months. Survival, Body Weights, and Feed Consumption: Survival of exposed males and females was similar to that of the controls. The mean body weights of rats in the 2,500 ppm groups were less than those of the controls beginning at week 2 of the study. Feed consumption by exposed groups was similar to that by the controls throughout the study. Pathology Findings: Three Zymbal's gland carcinomas were observed in female rats exposed to 2,500 ppm. This incidence exceeded the range observed for current historical controls and was considered an equivocal finding. At the 6- and 12 month interim evaluations and at 2 years, emodin-related increases in the incidences of renal tubule hyaline droplets occurred in all exposed groups. The incidences of renal tubule pigmentation were significantly increased in all exposed groups of males at 2 years. There were negative trends in the incidences of mononuclear cell leukemia in male and female rats, and the incidences in the 2,500 ppm groups were significantly decreased. In females exposed to 2,500 ppm, the incidence was below the historical control range; the incidence in males exposed to 2,500 ppm was at the lower end of the historical control range. 2-YEAR STUDY IN MICE: Groups of 60 male mice were fed diets containing 0, 160, 312, or 625 ppm emodin (equivalent to average daily doses of approximately 15, 35, or 70 mg/kg) for 105 weeks. Groups of 60 female mice were fed diets containing 0, 312, 625, or 1,250 ppm emodin (equivalent to average daily doses of approximately 30, 60, or 120 mg/kg) for 105 weeks. Ten male and ten female mice from each group were necropsied at 12 months. Survival, Body Weights, and Feed Consumption Survival and mean body weights of exposed males and females were similar to those of the controls. No differences in feed consumption were noted between exposed and control groups. Pathology Findings: Low incidences of renal tubule adenoma and carcinoma occurred in exposed male mice; these incidences included one carcinoma each in the 312 and 625 ppm groups. Renal tubule neoplasms are rare in male mice, and their presence in these groups suggested a possible association with emodin exposure. At the 12-month interim evaluation, the severity of nephropathy was slightly increased in males exposed to 625 ppm. Also at 12 months, the severity of nephropathy increased from minimal in the lower exposure groups to mild in females exposed to 1,250 ppm; the incidence in this group was significantly increased compared to the control group. At 2 years, the severities of nephropathy were slightly increased in males exposed to 625 ppm and females exposed to 1,250 ppm. The incidences of nephropathy were significantly increased in all exposed groups of females. At the 12-month interim evaluation, the incidences of renal tubule pigmentation were significantly increased in all exposed groups of males and in females exposed to 625 or 1,250 ppm. The severities increased with increasing exposure concentration. At 2 years, the incidences of renal tubule pigmentation were significantly increased in all exposed groups; severities increased with increasing exposure concentration. GENETIC TOXICOLOGY: Emodin was mutagenic in Salmonella typhimurium strain TA100 in the presence of S9 activation; no mutagenicity was detected in strain TA98, with or without S9. Chromosomal aberrations were induced in cultured Chinese hamster ovary cells treated with emodin, with and without S9. Three separate in vivo micronucleus tests were performed with emodin. A male rat bone marrow micronucleus test, with emodin administered by three intraperitoneal injections, gave negative results. Results of acute-exposure (intraperitoneal injection) micronucleus tests in bone marrow and peripheral blood erythrocytes of male and female mice were negative. In a peripheral blood micronucleus test on mice from the 14-week study, negative results were seen in male mice, but a weakly positive response was observed in similarly exposed females. CONCLUSIONS: Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity of emodin in male F344/N rats exposed to 280, 830, or 2,500 ppm. There was equivocal evidence of carcinogenic activity of emodin in female F344/N rats based on a marginal increase in the incidence of Zymbal's gland carcinoma. There was equivocal evidence of carcinogenic activity of emodin in male B6C3F1 mice based on a low incidence of uncommon renal tubule neoplasms. There was no evidence of carcinogenic activity of emodin in female B6C3F1 mice exposed to 312, 625, or 1,250 ppm. Exposure of rats to emodin resulted in increased incidences of renal tubule hyaline droplets and pigmentation in males, increased incidences of renal tubule hyaline droplets in females, and increased severities of renal tubule pigmentation in males and females. Emodin exposure resulted in increased incidences of renal tubule pigmentation in male and female mice and increased incidences of nephropathy in female mice. Incidences of mononuclear cell leukemia decreased in male and female rats exposed to 2,500 ppm. Synonyms: Archin; C.I. 75440; C.I. Natural Green 2; C.I. Natural Yellow 14; emodol; frangulic acid; frangula emodin; 6-methyl- 1,3,8-trihydroxyanthraquinone; Persian Berry Lake; rheum emodin; schuttgelb; 1,3,8-trihydroxy-6-methyl-9,10- anthracenedione; 1,3,8 trihydroxy-6-methylanthraquinone; 4,5,7-trihydroxy-2-methylanthraquinone. PMID- 12563348 TI - NTP Toxicology and Carcinogenesis Studies of Gallium Arsenide (CAS No. 1303-00-0) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Gallium arsenide is used primarily to make light- emitting diodes, lasers, laser windows, and photodetectors and in the photoelectronic transmission of data through optical fibers. Gallium arsenide was nominated for study because of its widespread use in the microelectronics industry, the potential for worker exposure, and the absence of chronic toxicity data. Male and female F344/N rats and B6C3F1 mice were exposed to gallium arsenide particles (greater than 98% pure; mass median aerodynamic diameter = 0.8 to 1.0 &mgr;m) by inhalation for 16 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, and the frequency of micronuclei was determined in the peripheral blood of mice exposed to gallium arsenide for 14 weeks. 16-DAY STUDY IN RATS: Groups of five male and five female rats were exposed to particulate aerosols of gallium arsenide with a mass median aerodynamic diameter of approximately 1 &mgr;m at concentrations of 0, 1, 10, 37, 75, or 150 mg/m(3) by inhalation, 6 hours per day, 5 days per week, for 16 days. All rats survived to the end of the study. The final mean body weights of all exposed groups of males and females were similar to those of the chamber controls. Compared to chamber controls, the liver and lung weights of males exposed to 1 mg/m(3) or greater and females exposed to 10 mg/m(3) or greater were increased; the thymus weights of all exposed groups of males were decreased. Gallium arsenide particles were visible in the alveolar spaces and, to a lesser extent, within alveolar macrophages of exposed rats. Moderate proteinosis (surfactant mixed with small amounts of fibrin) and minimal histiocytic cellular infiltrate were observed in the alveoli of exposed males and females. Epithelial hyperplasia and squamous metaplasia of the larynx were observed primarily in males exposed to 150 mg/m(3). 16-DAY STUDY IN MICE: Groups of five male and four or five female mice were exposed to particulate aerosols of gallium arsenide with a mass median aerodynamic diameter of approximately 1 &mgr;m at concentrations of 0, 1, 10, 37, 75, or 150 mg/m(3) by inhalation, 6 hours per day, 5 days per week, for 16 days. The final mean body weights were similar among exposed and chamber control groups. Compared to chamber controls, the lung weights of males and females exposed to 10 mg/m(3) or greater were increased. Gallium ar senide particles were visible in alveolar spaces and macrophages in some mice exposed to 150 mg/m(3). Moderate proteinosis, mild epithelial hyperplasia, and histiocytic infiltration of the lung were observed in males and females exposed to 10 mg/m(3) or greater. In the larynx, mild squamous metaplasia was seen in mice exposed to 10 mg/m(3) or greater, and mild chronic inflammation occurred in mice exposed to 75 or 150 mg/m(3). 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed by inhalation to gallium arsenide particulate at concentrations of 0, 0.1, 1, 10, 37, or 75 mg/m(3), 6 hours per day, 5 days per week, for 14 weeks. All rats survived until the end of the study. The final mean body weight and body weight gain of males exposed to 75 mg/m(3) were significantly less than those of the chamber controls. Hematology and clinical chemistry results indicated that exposure to gallium arsenide induced a microcytic responsive anemia with an erythrocytosis and increased zinc protoporphyrin/heme ratios in exposed groups of rats. There were also increases in platelet and neutrophil counts, a transient decrease in leukocyte counts, and increases in the serum activities of alanine aminotransferase and sorbitol dehydrogenase. These changes were of greater magnitude in male rats. The lung weights of all exposed groups of rats were increased, while testis, cauda epididymis, and epididymis weights of males exposed to 37 or 75 mg/m(3) were generally less than those of chamber controls. Total spermatid heads and spermatid counts were significantly decreased in males exposed to 75 mg/m(3), while epididymal spermatozoa motility was significantly reduced in males ees exposed to 10 mg/m(3) or greater. Gallium arsenide particles were visible in alveolar spaces and macrophages in the lungs of exposed rats. Minimal to marked proteinosis and minimal histiocytic cellular infiltration of the alveoli were observed in all exposed groups; minimal squamous metaplasia in the larynx and lymphoid cell hyperplasia of the mediastinal lymph node were observed in some males and females exposed to 37 or 75 mg/m(3). Exposure-related increases in the incidences of plasma cell hyperplasia of the mandibular lymph node, testicular atrophy, epididymal hypospermia, bone marrow hyperplasia (males), and hemosiderosis in the liver were observed in the 37 and 75 mg/m(3) groups. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were exposed by inhalation to gallium arsenide particulate at concentrations of 0, 0.1, 1, 10, 37, or 75 mg/m(3), 6 hours per day, 5 days per week, for 14 weeks. One female mouse exposed to 75 mg/m(3) died before the end of the study. Final mean body weights and body weight gains of males in the 75 mg/m(3) group were signifi cantly less than the chamber controls. Hematology and clinical chemistry results indicated that exposure to gallium arsenide affected the circulating erythroid mass and induced a microcytic responsive anemia with an erythrocytosis and increased zinc protoporphyrin/heme ratios in male and female mice. There were also increases in platelet and neutrophil counts. Compared to the chamber controls, the lung weights of males exposed to 1 mg/m(3) or greater and females exposed to 10 mg/m(3) or greater were increased. Testis, cauda epididymis, and epididymis weights, total spermatid heads, spermatid counts, and concentration and motility of epididymal spermatozoa were generally decreased. Gallium arsenide particles were visible in alveolar spaces and macrophages in the lungs of mice exposed to 1 mg/m(3) or greater. Mild to marked proteinosis, histiocytic infiltration, and epithelial hyperplasia were observed in the alveoli of males and females exposed to 1 mg/m(3) or greater. Minimal to mild suppurative inflammation and granuloma in the lung and squamous metaplasia in the larynx were present in males and females exposed to 10 mg/m(3) or greater. Min imal hyperplasia was observed in the tracheobronchial lymph node of males exposed to 10 mg/m(3) or greater and females exposed to 37 or 75 mg/m(3). Exposure- related increases in the incidences of testicular atrophy, epididymal hypospermia, hematopoietic cell proliferation of the spleen, and hemosiderosis of the liver and spleen were observed in groups of male and female mice exposed to 10 mg/m(3) or greater. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed by inhalation to gallium arsenide particulate at concentrations of 0, 0.01, 0.1, or 1.0 mg/m(3), 6 hours per day, 5 days per week, for 105 weeks. Survival and Body Weights: Survival of exposed male and female rats was similar to the chamber controls. Mean body weights of males exposed to 1.0 mg/m(3) were generally less than those of the chamber controls throughout the study; females exposed to 1.0 mg/m(3) had slightly lower mean body weights during the second year. Pathology Findings: Compared to the chamber controls, the incidences of alveolar/bronchiolar neoplasms were significantly increased in females exposed to 1.0 mg/m(3) and exceeded the historical control ranges. Exposure-related nonneoplastic lesions in the lungs of male and female rats included atypical hyperplasia, alveolar epithelial hyperplasia, chronic active inflammation, proteinosis, and alveolar epithelial metaplasia. In the larynx of males exposed to 1.0 mg/m(3), the incidences of hyperplasia, chronic active inflammation, squamous metaplasia, and hyperplasia of the epiglottis were significantly increased. The incidences of benign pheochromocytoma of the adrenal medulla occurred with a positive trend in female rats, and the incidence was significantly increased in the 1.0 mg/m(3) group and exceeded the historical control range. The incidence of mononuclear cell leukemia was significantly increased in females exposed to 1.0 mg/m(3) and exceeded the historical control range. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed by inhalation to gallium arsenide particulate at concentrations of 0, 0.1, 0.5, or 1.0 mg/m(3), 6 hours per day, 5 days per week, for 105 (males) or 106 (females) weeks. Survival and Body Weights: Survival of male and female mice was similar to the chamber controls. Mean body weights of exposed groups of males were similar to those of the chamber controls throughout the study; mean body weights of exposed groups of females were greater than those of the chamber controls from week 13 until the end of the study. Pathology Findings: Exposure-related nonneoplastic lesions in the lung of all groups of exposed mice included suppurative focal inflammation, chronic focal inflammation, histiocyte cellular infiltration, alveolar epithelial hyperplasia, and proteinosis. Increased incidences of minimal lymphoid hyperplasia of the tracheobronchial lymph node occurred in mice exposed to 1.0 mg/m(3) and in 0.5 mg/m(3)mg/m(3) males. GENETIC TOXICOLOGY: Gallium arsenide was not mutagenic in several strains of Salmonella typhimurium, with or without S9 metabolic activation enzymes, and no increase in the frequency of micronucleated erythrocytes was observed in peripheral blood of male or female mice exposed to gallium arsenide by inhalation for 14 weeks. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was no evidence of carcinogenic activity of gallium arsenide in male F344/N rats exposed to 0.01, 0.1, or 1.0 mg/m(3). There was clear evidence of carcinogenic activity in female F344/N rats based on increased incidences of benign and malignant neoplasms in the lung. Increased incidences of benign neoplasms of the adrenal medulla and increased incidences of mononuclear cell leukemia were also considered to be exposure related. There was no evidence of carcinogenic activity in male or female B6C3F1 mice exposed to 0.1, 0.5, or 1.0 mg/m(3). Exposure to gallium arsenide caused a spectrum of nonneoplastic lesions in the lung of rats and mice, the larynx of male rats and hyperplasia of the tracheobronchial lymph node in mice. Synonym: Gallium monoarsenide. PMID- 12563349 TI - NTP Toxicology and Carcinogenesis Studies of Methyleugenol (CAS NO. 93-15-2) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Methyleugenol is used as a flavoring agent in jellies, baked goods, nonalcoholic beverages, chewing gum, candy, pudding, relish, and ice cream. It is also used as a fragrance in perfumes, creams, lotions, detergents, and soaps. Methyleugenol has also been used as an insect attractant in eradication programs and as an anesthetic in rodents. Methyleugenol was nominated for testing because of its widespread use and because of its structural resemblance to safrole, a known carcinogen, and isosafrole and estragole. Male and female F344/N rats and B6C3F1 mice received methyleugenol (approximately 99% pure) in 0.5% methylcellulose by gavage for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and mouse peripheral blood erythrocytes. 14-WEEK STUDY IN RATS: Groups of 9 or 10 male and 10 female F344/N rats were administered 0, 10, 30, 100, 300, or 1,000 mg methyleugenol/kg body weight in 0.5% methylcellulose by gavage 5 days per week for 14 weeks. A water control group of 10 male and 10 female rats received deionized water by gavage. All rats survived until the end of the study. The final mean body weights of 300 and 1,000 mg/kg males and of all dosed groups of females were significantly less than those of the vehicle controls. Erythrocyte microcytosis was demonstrated by decreased mean cell volumes in 300 mg/kg males and 1,000 mg/kg males and females. There was evidence of a thrombocytosis at all time points, demonstrated by increased platelet counts in the 100 mg/kg or greater groups. The serum activities of alanine aminotransferase and sorbitol dehydrogenase were increased in the 100 mg/kg or greater rats at various time points, suggesting hepatocellular injury. Additionally, bile acid concentrations were generally increased in the 300 and 1,000 mg/kg groups at all time points, consistent with cholestasis or altered hepatic function. A hypoproteinemia and hypoalbuminemia, evidenced by decreased total protein and albumin concentrations, occurred in rats in the 300 and 1,000 mg/kg groups at all time points. Liver weights of 100, 300, and 1,000 mg/kg males and 300 and 1,000 mg/kg females and testis weights of 1,000 mg/kg males were significantly increased. Increased incidences of liver lesions occurred in 300 and 1,000 mg/kg males and females and hepatocellular adenoma occurred in one 1,000 mg/kg male. The incidences of atrophy and chronic inflammation of the mucosa of the glandular stomach were significantly increased in rats administered 300 or 1,000 mg/kg. Increased incidences of adrenal gland cortical hypertrophy and/or cytoplasmic alteration in the submandibular gland occurred in the 100 mg/kg or greater groups. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice received methyleugenol in 0.5% methylcellulose by gavage at doses of 0, 10, 30, 100, 300, or 1,000 mg/kg, 5 days per week for 14 weeks. A water control group of 10 male and 10 female mice received deionized water by gavage. All but one male and all females receiving 1,000 mg/kg died before the end of the study. The mean body weight gains of mice in the 300 mg/kg groups were significantly less than those of the vehicle controls. The only clinical finding was toxicity manifested as generalized morbidity in mice administered 1,000 mg/kg. Liver weights of 30, 100, and 300 mg/kg males and of 300 mg/kg females were significantly increased. Male mice administered 10 or 30 mg/kg had significantly lower cauda epididymis, epididymis, and testis weights; males receiving 100 mg/kg had significantly lower spermatozoal concentrations. Increased incidences of liver lesions occurred in 1,000 mg/kg males and 300 and 1,000 mg/kg females. The incidences of lesions of the glandular stomach were increased in one or more groups administered 30 mg/kg or greater. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats received methyleugenol in 0.5% methylcellulose by gavage at doses of 37, 75, or 150 mg/kg, 5 days per week for 105 weeks; groups of 60 male and 60 female rats received the 0.5% me60 female rats received the 0.5% methylcellulose vehicle only. Stop exposure groups of 60 male and 60 female rats received 300 mg/kg in 0.5% methylcellulose by gavage for 52 weeks followed by just the 0.5% methylcellulose vehicle for the remaining 53 weeks of the study. Special study groups of 10 male and 10 female rats administered 36, 75, 150, or 300 mg/kg were designated for toxicokinetic studies. Survival and Body Weights: All 150 and 300 mg/kg males died before the end of the study, and survival of 150 mg/kg females was slightly less than that of the vehicle controls. Mean body weights of all dosed groups of rats were less than those of the vehicle controls throughout most of the 2-year study. Pathology Findings: Chemical-related liver neoplasms occurred in all dosed groups of rats and included hepatocellular adenoma, hepatocellular carcinoma, hepatocholangioma, and hepatocholangiocarcinoma; at 2 years, there were positive trends in the incidences of hepatocellular adenoma, carcinoma, and adenoma or carcinoma (combined) in core study rats and in the numbers of rats with multiple liver neoplasms. Nonneoplastic lesions included eosinophilic and mixed cell foci, hepatocellular hypertrophy, oval cell hyperplasia, cystic degeneration, and bile duct hyperplasia (females); the incidences of these lesions in dosed groups of male and female rats were increased at 6 months, 12 months, and/or 2 years. Chemical-related neoplasms and nonneoplastic lesions of the glandular stomach included benign and malignant neuroendocrine tumors in the 150 and 300 mg/kg groups and females in the 75 mg/kg group. In all dosed groups of rats at all time points, the incidences of mucosal atrophy were significantly greater than in the vehicle controls. Neuroendocrine cell hyperplasia was observed in females at 6 months and males and females at 12 months and at 2 years. In core study female rats, there was a positive trend in the incidences of squamous cell papilloma or carcinoma (combined) of the forestomach, and the incidence in the 150 mg/kg group exceeded the historical control range. The incidences of renal tubule proliferative lesions in male rats were suggestive of a neoplastic effect in the kidney. Therefore, additional step sections of the kidneys of male rats were prepared. The incidences of renal tubule hyperplasia and adenoma in the extended evaluation and the combined incidences of standard and step sections in the 75, 150, and 300 mg/kg groups were greater than those in the vehicle controls. The incidences of nephropathy were increased in all dosed groups of females, and the increase was significant in the 300 mg/kg group. In dosed groups of male rats, there was a positive trend in the incidences of malignant mesothelioma, and the incidences were significantly greater in 150 and 300 mg/kg males than in the vehicle controls. The incidences of mammary gland fibroadenoma in 75 and 150 mg/kg males were significantly increased. The incidences of fibroma of the subcutaneous tissue in 37 and 75 mg/kg males and the combined incidences of fibroma or fibrosarcoma in 37, 75, and 150 mg/kg males were significantly increased. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice received methyleugenol in 0.5% methylcellulose by gavage at doses of 0, 37, 75, or 150 mg/kg for 105 weeks. Special study groups of 10 male and 10 female mice administered 37, 75, or 150 mg/kg were designated for toxicokinetic studies. Survival and Body Weights: Survival of all dosed groups of male mice was similar to that of the vehicle controls. Survival of dosed groups of females was significantly less. Mean body weights of dosed mice were generally less than those of the vehicle controls throughout the studies. Pathology Findings: Chemical-related increases in the incidences of liver neoplasms and nonneoplastic lesions in mice included hepatocellular adenoma and carcinoma, hepatoblastoma, hepatocholangiocarcinoma, eosinophilic foci, oval cell hyperplasia, bile duct hyperplasia, hemosiderin pigmentation, chronic active inflammation, and hematopoietic cell proliferation. In all dosed groups ofmales and females, the incidences of hepatocellular neoplasms and the multiplicity of neoplasms were generally greater than in the vehicle controls. The incidences of hepatoblastoma were significantly increased in all dosed groups of females and slightly increased in 150 mg/kg males. Hepatocholangiocarcinoma was observed in 150 mg/kg females. The incidences of eosinophilic foci, oval cell hyperplasia, portal hypertrophy, hepatocyte necrosis, hematopoietic cell proliferation, bile duct hyperplasia, and hemosiderin pigmentation were significantly increased in two or more dosed groups of male and/or female mice. The incidences of glandular ectasia, mucosal atrophy, chronic active inflammation, epithelial hyperplasia, and neuroendocrine cell hyperplasia of the glandular stomach were increased in one or more dosed groups of male and female mice. In addition, malignant neuroendocrine tumors were observed in the glandular stomach of two 150 mg/kg male mice; one male in this group had a carcinoma. TOXICOKINETIC STUDIES: Methyleugenol is rapidly absorbed following oral administration to rats and mice. The kinetic data are consistent with rapid clearance from the blood, metabolism in the liver, and excretion of the parent and various metabolites in the urine. GENETIC TOXICOLOGY: Methyleugenol was not mutagenic in S. typhimurium strain TA98, TA100, TA1535, or TA1537, with or without exogenous metabolic activation (S9). In cytogenetic tests with cultured Chinese hamster ovary cells, methyleugenol induced sister chromatid exchanges in the presence of S9, but no induction of chromosomal aberrations was noted in cultured Chinese hamster ovary cells following exposure to methyleugenol, with or without S9. In vivo, no increase in the frequency of micronucleated normochromatic erythrocytes was seen in male or female mice administered methyleugenol by gavage for 14 weeks. PHYSIOLOGICALLY BASED PHARMACOKINETIC MODEL: A physiologically based pharmacokinetic (PBPK) model resulting from intravenous and oral exposure was created to characterize tissue concentrations of methyleugenol in rats and mice. Data used to create the model were obtained from the literature or from current studies. The primary conclusions that can be reached from the PBPK model are: 1) absorption of oral doses of methyleugenol in rats and mice is rapid and complete, 2) distribution of methyleugenol to tissues is not hampered by capillary permeability, and 3) metabolism of methyleugenol is saturable and must have some extrahepatic component in the mouse. Model-based plasma methyleugenol concentrations were not found to be good dosimeters for evaluating neoplasm dose-response data. CONCLUSIONS: Under the conditions of these 2-year gavage studies, there was clear evidence of carcinogenic activity of methyleugenol in male and female F344/N rats based on the increased incidences of liver neoplasms and neuroendocrine tumors of the glandular stomach in male and female rats and the increased incidences of kidney neoplasms, malignant mesothelioma, mammary gland fibroadenoma, and subcutaneous fibroma and fibroma or fibrosarcoma (combined) in male rats. A marginal increase in the incidence of squamous cell neoplasms of the forestomach may have been related to methyleugenol administration in female rats. There was clear evidence of carcinogenic activity of methyleugenol in male and female B6C3F1 mice based on the increased incidences of liver neoplasms. Neuroendocrine tumors of the glandular stomach in male mice were also considered related to methyleugenol administration. In male and female rats and mice, methyleugenol administration caused significant increases in nonneoplastic lesions of the liver and glandular stomach. Synonyms: 1-Allyl-1,2-dimethoxybenzene; 4-allylveratrole; 4-allyl-1,2-dimethoxy-benzene; 1,2-dimethoxy-4-allylbenzene; 3,4 dimethoxyallylbenzene; ENT 21040; 1-(3,4-dimethoxyphenyl)-2-propene; eugenol methyl ether; 1,3,4-eugenol methyl ether; veratrole methyl ether. PMID- 12563350 TI - [The course of cancer mortality in Hungary between 1975 and 2001]. AB - The course of cancer mortality in this country between 1975 and 2001 was analysed solely with mathematical methods using the mortality data provided by the Central Statistical Office. Mortality data were studied according to patient s sex and tumour localisation and in relation to total cancer mortality. The increase and decrease in cancer mortality were found to differ by sex and tumour localisation: e.g. death rate caused by cancers of the oral cavity showed low deviation with an even increase just like the mortality caused by colorectal cancer, the latter, however, was steeper with men. In case of melanoma higher deviation was associated with increased mortality, again at a higher rate with men. Dying of testicular cancer and of gastric cancer in either sex showed decreasing tendencies. Lung cancer mortality assumed different patterns in the two genders: with men it kept increasing at an even pace until 1994 then the increase stopped. With women, however, the increase since 1985 was steeper than earlier. The breast cancer mortality rates can also be divided into two periods. There was an even rise until 1994 followed by stagnation. As to the total cancer mortality values, the authors state that the rhythm of increase during the first 20 years of the study period had changed, the steepness of trends in the last seven years can be expressed in a small positive number not differing from zero at significant level which means that the increase in cancer mortality has stopped. PMID- 12563351 TI - [Report on the first year of the activity of the National Oncological RD Consortium]. AB - The project focuses on cancer types with outstanding publich health importance (breast-, colorectalhead and neck cancers and childhood tumors). Epidemiological studies revealed significant regional differences in the mobidity/mortality of these cancer types in Hungary. Molecular epidemiological studies revealed characteristic BRCA1 mutation patterns of familiar breast cancer and DNA repair enzyme polymorphism in head and neck cancer. New methods have been developed for the screening (lactoferrin), prognostication (c-met expression) or the prediction of therapeutic sensitivity (TS expression) of colorectal cancer. In the pediatric oncology program alternative way of MRD monitoring (WT1 expression) and a potential new therapeutic modality (IFN-alpha) of ALL was developed. Experimental studies demonstrated that the tumoral matrix significantly influences the effects of the classic chemotherapeutic agents. We have identified several genes the expression of which could serve efficiently as markers or targets for therapy of the progression of melanoma (alphaIIbbeta3 integrin, CD44v3 and decorin proteoglycans, AMF receptor). PMID- 12563352 TI - [Evolution of adjuvant chemotherapy of breast cancer from Bonadonna to the taxanes]. AB - The author summarizes the progress of adjuvant chemotherapy of breast cancer from the classical Bonadonna-type CMF over the anthracyclines to the taxanes. The CMF regimen represented the prototype of combination chemotherapy which significantly improved early and long term results. After 20 years the patients given adjuvant combination chemotherapy with CMF had significantly better rates of relapse-free survival (p<0.001) and overall survival (p=0.03) compared with no chemotherapy. 6 cycles of CMF was the gold standard of adjuvant chemotherapy in breast cancer for decades. The Milan research group decided in the early 1980s to challange this popular CMF combination by introducing doxorubicin within the adjuvant program. Compared with standard CMF, anthracyclin-containing regimens reduced the annual risk of recurrence by 12% and the annual risk of death by 11%, equating to a 3.2% absolute reduction in recurrence and a 2.7% absolute reduction in mortality at 5 years. This small but real difference seen with regimens containing three or more agents (e.g. CEF and CAF, FAC, FEC, etc.), whereas 4 cycles of 2-drug regimens (e.g. AC or EC) appears to be equivalent to 6 cycles of CMF. Among the novel chemotherapeutic drugs introduced in the 1990s the taxanes have emerged as the most powerful compounds in breast cancer. Several large, adjuvant clinical trials are currently ongoing or have recently completed accrual. The available results from innumerable clinical studies are still inconclusive and do not support the routine use of taxanes in the adjuvant setting - with the exception of the BCIRG 001 docetaxel trial, in which significant improvement was documented in disease free survival with 6 x TAC compared with 6 x FAC (82% vs 74%). Studies on the effect of the new trastuzumab (an antibody against the extracellular domain of the HER2) in adjuvant setting was initiated in early 2000. The Herceptin adjuvant trial programme is extensive, involving more than 12,000 patients worldwide. This trials will potentially offer many women with HER2-positive disease the chance of improved survival. PMID- 12563353 TI - [Sentinel lymph node biopsy in Hungary. Results with a revolutionary new method in surgical oncology]. AB - This review deals with results attained by Hungarian authors in the field of sentinel nodes and presents the current status of sentinel lymphadenectomy in Hungary. After a short historical overview, results with melanoma and breast cancer are summarized, and feasibility studies on other possible sites (gastrointestinal tract, thyroid gland) are also mentioned. Pathological aspects are also dealt with in a separate section. The summary of these results suggests that Hungarian authors have investigated several facets of sentinel node biopsy, their results are in keeping with international results. Despite the favourable results, the method still needs time to be more widely accepted and reflected in the literature. PMID- 12563354 TI - [Radiotherapy and delayed breast reconstruction with implant: examination of compatibility]. AB - PURPOSE: Delayed breast reconstruction with implant is frequently used after mastectomy. However, given that many breast cancer patients receive adjuvant radiotherapy, the adaptation of this method raises questions concerning possible cumulative complications. In the present study the compatibility of post mastectomy radiotherapy and delayed breast reconstruction with implant was examined. PATIENTS AND METHODS: We evaluated a consecutive series of 66 invasive breast cancer patients who have received delayed breast reconstruction with implant after modified radical mastectomy between January 1997 and June 1999. The average time was 51 months from primary surgery to reconstruction. The median dose of loco-regional irradiation was 50 Gy. Grade III atrophic dermatitis was observed in none of the irradiated women. We identified two patient groups: 29 patients did not and 37 patients did receive post-mastectomy radiotherapy. The types and time of reconstruction related chronic complications (capsular contracture, defect of implant shell and skin necrosis) were recorded. Incidence of complications was estimated by the Kaplan-Meier method. Cancer related events were also studied. RESULTS: When expander was used, minor discomfort of the patients was common during the filling course. At four (11%) irradiated patients the expansion with expander was incomplete, due to severe pain. At a median follow up time of 53 months the incidence of capsular contracture was 24.1% without and 29.7% with radiotherapy (p=0.4121). The five-year estimated rate of late complications was 40% and 50%, respectively (relative risk: 1.29, 95% confidence interval: 0.58-2.89, p=0.5173). The position of implant had an impact on the incidence of capsular contracture: 46.2% with subcutaneous and 22.6% with submuscular position (p=0.0881). Four patients (6%) developed local relapse (three in the skin and one subcutaneous). All were treated with tumor excision without implant removal. CONCLUSION: Delayed breast reconstruction with implant after post-mastectomy radiotherapy can be offered to patients who are interested in breast reconstruction and had no severe late radiation skin toxicity. Post mastectomy radiotherapy does not significantly increase the risk of complications. The use of skin expander is less tolerated by irradiated patients. Submuscular position of implants moderates the risk of capsular contracture. PMID- 12563355 TI - [Serum levels of sex steroid and pituitary hormones in chronic alcoholics and head and neck cancer patients as compared to normal controls]. AB - Head and neck squamous cell carcinoma (HNSCC) is diagnosed mainly in male patients (more than 80% of the cases) with a history of smoking and heavy alcohol consumption. However, only a few percent of all alcoholics develop head and neck cancer. OBJECTIVE OF THE STUDY: to investigate the hormonal status in HNSCC patients as compared to healthy controls and alcoholic persons in order to find changes, if any, characteristic for cancer. METHOD: The liver function expressed by gamma-GT levels, the hypophysis gonadotrop hormone (FSH, LH, prolactin) and sex steroid hormone serum levels were examined in 130 male HNSCC patients, in 54 men with alcoholic liver disease but without any known cancer and in 56 healthy men as controls. RESULTS: When compared to the healthy controls, both alcoholics and tumor patients had abnormal liver function, testosterone, sex hormone binding globuline and prolactin levels, reflecting the presence of alcoholic liver disease in tumor patients as well. However, abnormally elevated circulating FSH (p<0.005) and LH (p<0.0003) levels were present only in the tumor patients. CONCLUSION: Sex steroid hormone abnormalities are common among head and neck cancer patients, mainly as results of the chronic alcoholic liver disease. Elevation of FSH and LH levels suggests a potential role of these hormones in the formation of head and neck cancer. The exact role of the hypothalamus-hypophysis liver axis in the biology of head and neck cancer requires further investigations. PMID- 12563356 TI - [Progression of leukoplakia is associated with changes in apoptotic and mitotic index as well as in p53 and Ki-67 expression]. AB - INTRODUCTION: Leukoplakia is the most frequent preblastomatous alteration in the oral cavity. Its potential for malignant transformation is unpredictable. The aim of the present study was to provide data to the prognosis and molecular genetic background of this disease. MATERIALS AND METHODS: Biopsy material from 15 leukoplakia patients and three oral squamous cell carcinoma patients treated at the Department of Periodontology, Semmelweis University were studied with histological and immunohistochemical methods. Hemotoxylin and eosin staining, Apop-Detect kit (for TUNEL reaction), immunohistochemical reactions for Ki-67 and p53 were applied. The severity of dysplasia, mitotic and apoptotic index and expression as well as distribution of Ki-67 and p53 were examined and related to the clinical appearance of leukoplakia. RESULTS: Mitotic and apoptotic index, Ki 67 expression increased significantly in parallel with the severity of dysplasia and also with the clinical stage (homogenous, nodular erythroleukoplakia). Positivity and intracellular localisation of mutant p53 varied according to the clinical forms of leukoplakia. Homogenous and nodular forms showed cytoplasmic while erythroleukoplakia and carcinoma were characterized by nuclear positivity. CONCLUSION: Increased mitotic, apoptotic and Ki-67 index may indicate unfavourable prognosis of leukoplakia. The expression of Ki-67 and p53 in various forms of leukoplakia varies in parallel with the severity of leukoplakia. PMID- 12563357 TI - [Lung resistance protein analysis in testicular cancer]. AB - Germ cell testicular cancers are well-curable neoplasms, because total remission can be achieved in about 80% of the cases. However, 15-20% of the patients die due to drug resistance (DR). A number of mechanisms of the multidrug resistance phenotype are known, including MDR/P-glycoprotein (P-gp) and the so-called multidrug resistance associated protein (MRP). Lung Resistance Protein (LRP) is an ATP dependent membrane transporter protein associated with MDR. In our present work we studied the expression of LRP in testicular cancers. LRP expression was determined by immunohistochemistry (IH), Western blot (WB) and RT-PCR techniques. Clinical resistance was defined in accordance with the clinical oncologic rules. In 29 (41%) of 70 primary testicular tumours and in 22 (63%) of 35 cases, elevated LRP levels were established by IH and WB, respectively. In the latter 63%, the LRP mRNA levels were elevated as well. Six cases of the 15 seminomas and 23 cases of the nonseminomatous germ cell tumours (NSGCT) proved to be positive. No relationship was demonstrated between LRP expression and the stage of the disease. Despite the LRP positivity of 6 tumour samples, all of the seminomas proved sensitive. Of the 39 sensitive NSGCT, 27 cases were LRP-negative, whereas 11 tumour samples of 16 patients belonging to the resistant group proved LRP positive (p=0.04). The authors concluded that the expression of LRP is responsible for clinical drug resistance in non-seminomatous testicular cancer patients. PMID- 12563358 TI - [Lingual thyroid: a rare disease resembling base of tongue neoplasm]. AB - The ectopic thyroid gland occurring in the midline of the base of tongue is a rare developmental anomaly. It may cause differential diagnostic problems with real malignant tumor. Symptoms, if where are any: foreign-body-feeling, swallowing difficulties, dyspnea, articulation disorders, bleeding and hypothyroidism, but in many cases the diagnosis is accidental. We describe two cases of lingual thyroid gland operated in our department, and discuss the present trends of the treatment of this disease. We agree with most of the authors that only cases presenting with symptoms should be operated, and if possible the normal thyroid tissue should be replaced into the body. However, all discovered cases have to be followed to avoid late hormonal disturbances. PMID- 12563359 TI - [Renal and ureter complications from the treatment of Hodgkin's disease]. AB - PURPOSE: The authors analysed renal and ureter complications in Hodgkin's disease patients after treatment. PATIENTS AND METHODS: We examined retrospectively 512 primary treated and followed up patients with Hodgkin's disease. RESULTS: We observed renal, ureter or bladder complications after irradiation in 16 cases (3.125%). Five patients had injured left kidney, out of them only two had pyuria or proteinuria and 4 received radiotherapy and chemotherapy as well. We observed complications of both kidneys in 7 patients. Four patients had pyelonephritis or cystitis. We did not find severe cystitis in patients treated wsith cyclophosphamide. CONCLUSIONS: Acute and chronic irradiation nephropathy are rather anatomic than functional lesions. Planning, dose and period of the radiotherapy, irradiation volume play parts in the development of complications. Prior chemotherapy increases incidence of irradiation nephropathy. These rare complications of Hodgkin's disease are usually avoided with the use of modern radiotherapy apparatus and the up to date treatment methods. PMID- 12563360 TI - [Incidence of valvulopathy in Hodgkin-disease patients in long-term complete remission]. AB - AIM: To study the occurrence of valvulopathies after treatment in Hodgkin's disease patients. PATIENTS AND METHODS: 124 Hodgkin's disease patients in complete remission for at least 1 year were echocardiographically examined. RESULTS: Abnormal finding was observed in 48/124 (38.4%) of patients, all of them presented with regurgitation, no stenosis was observed. Regurgitation of grade I or II was recorded in most cases. We have found single valvulopathy in 25/48 (52.1%) of patients, and multiple valvulopathy in 23/48 (47.9%) of patients. In most cases (78.7%) the valvulopathy was detected in left heart. Among these 48 patients the ratio of females was significantly higher than those of males, and also the ratio of the patients in early phase compared with those in late phase. We could detect vitium mostly in those patients who had mediastinal irradiation. The combined treatment, including anthracycline therapy, did not increase the frequency of vitium. CONCLUSION: The occurrence of valvulopathy is frequent in Hodgkin's disease patients, particularly in patients treated by radiation. This is the reason why Hodgkin's disease patients should be examined regularly with echocardiography. These patients with valvulopathy need treatment adjusted to their state of health, and where possible, the complications should be prevented. In the future, when planning of the radiotherapy mediastinal irradiation of the Hodgkin's disease patients their heart protection from radiation should be taken into account. PMID- 12563361 TI - [The role of adjuvant clodronate therapy for prevention of bone metastasis in breast cancer]. AB - Bisphosphonates are effective against increased bone resorption because they inhibit osteoclast activity. The use of these drugs is well established for the treatment of metastatic breast and other cancers; they reduce skeletal complications, hypercalcemia, bone pain, and metastatic progression and they can improve the overall survival and quality of life. Preclinical observations and early clinical data indicate that early bisphosphonate treatment reduces the incidence and the extent of newly developed metastases in breast cancer. There is considerable interest in determining whether bisphosphonate treatment is to prevent the incidence of bone metastases and associated complications. To date three randomized, controlled clinical trials have examined the effect of long term use of clodronate (1600 mg/d po.) on the incidence of bone metastases, other metastases, the survival of patients, and the side effects of the study drug as well. All the trials have observed significant reduction of the occurrence of bone metastases, although this reduction was significant only during the medication period. One of the trials mentioned have shown an unexpected reduction in non-osseous metastases, and two of them have revealed significant improvements in the death rates. These promising results need further evaluation by large clinical trials with longer treatment periods to establish the clinical role of adjuvant bisphosphonate treatment of primary breast cancer. PMID- 12563362 TI - [The treatment of bone metastases with bisphosphonates--the patients' view]. AB - The Hungarian bisphosphonate market has been increasing for years; last year the number of patients was approximately 3200-3500. We decided to start a research among patients having malignant disease with bone metastases, in order to find out how patients evaluate the drugs, how they are informed and what is the role of doctors and nurses in compliance. Nearly 300 patients filled questionnaires and we have made 16 in-depth interviews. The average age of patients was 57 years. More than 60% of patients were younger than 60 and the male-female ratio was 1:2. We found that more than one quarter of the patients arrived to their doctor from farther than 50 km and 70% of them meet their doctor at least once a month. The results showed that 95% of patients would prefer oral treatment (tablets or capsules), and 75% of the patients would choose tablets, if they were asked. Most of the patients wanted to be informed primarily by doctors. Nurses were the hostesses of emotional problems. Doctor-patient relationship was characterised by paternalism. Female patients were more open to nurses, they talked over smaller problems emerging during the treatment and the same occurred with some male patients, too. Only 5% of patients received the treatment of their choice. Patients want to be involved more actively in the planning of their treatment process than doctors think, they expect that doctors should prefer their interest. Cancer patients are frequently underinformed and they expect more help to solve their psychological problems. PMID- 12563363 TI - Relapsing polychondritis: commentary. PMID- 12563364 TI - [Surgical treatment of the impingement syndrome and of the rotator cuff tears: personal experience in 134 cases]. AB - The time-course covered by the original definition of scapulo-humeral periarthritis suggested by Duplay through the more recent term of subacromial impingement syndrome coined by Neer, follows the identification of the pathogenetic mechanisms leading to chronic subacromial impingement and degenerative tears of the rotator cuff. The Authors recall the functional anatomic development evolution of the shoulder and the disequilibrium between the depressor and the elevator muscles which may promote the chronic friction against the acromion. However, the actual pathogenesis of the impingement still remains controversial. We evaluated 134 patients (81F, 53M, mean age 56.4 years) with chronic subacromial impingement syndrome. In 92 cases (69%) non traumatic tears of the rotator cuff were also present as confirmed by ultrasonography in 94 cases, CT in18 cases and magnetic resonance in 102 cases. The different tear patterns of the rotator cuff were classified as suggested by Ellman (L shaped, L reverse, triangular, trapezoidal and massive with retraction) and clinical results were analyzed following Sahlstrand clinical criteria and Costant numerical scale. According to the anatomical damage, patients were divided into those with impingement without severe cuff tendinopathy (42 cases), those with rotator cuff tears without loss of motion of the shoulder (32 cases) and those with cuff tears and loss of active motion (60 cases). The different surgical techniques and rehabilitation procedures after surgery are also reported. After a mean follow-up of 1.3 years (range 8 months - 2 years), good or excellent results were obtained in 91% of the patients without rotator cuff tears, in 87% of the patients with tears but without loss of motion and in 75% of the cases with loss of active motion. Our data demonstrate that in the majority of patients with chronic impingement syndrome and rotator cuff tears, surgical treatment shows high success rates. When surgery is associated with a careful rehabilitation programme it may frequently allow patients to return to their baseline function. PMID- 12563365 TI - [Hepatitis C virus infection and arthritis. A clinico-serological investigation of arthritis in patients with or without cryoglobulinemic syndrome]. AB - OBJECTIVE: To compare the clinico-serological features of arthritis from two HCV+ patient groups characterized by mixed cryoglobulinemia (MC) or chronic hepatitis (CH). METHODS: We retrospectively studied 157 MC patients (119 females, mean age 64.8 yrs, range 36-88) and 155 CH patients (103 females, mean age 58.8 yrs, range 30-81). Patients with HBV and/or HIV co-infections and a follow-up shorter than 1 year were excluded. MC was classified according to standard criteria, while CH classification was based on Desmet's criteria. RESULTS: No differences in epidemiology were demonstrated between the two series of patients. Although significantly prevalent in MC patients, extra-hepatic manifestations including nephropathy, neuropathy, pneumopathy, mixed cryoglobulins, RF positivity and hypocomplementemia were detected in both patient groups. Arthritis was present in 23 CH (15%) and 12 MC (8%) patients. A symmetrical polyarthritis was observed in 87% of 23 CH patients, while cryoglobulinemic arthritis was invariably asymmetrical and pauciarticular. Unlike MC patients, who had a constantly non erosive arthritis, in 7/23 CH patients arthritis was erosive. Steroids and/or hydroxycloroquine or D-penicillamine were safe and useful in controlling the arthritis over the short-medium time, although clinical response was more evident in MC patients. Instead, in 5/23 CH and 3/12 MC patients, interferon-alpha treatment was able to trigger or exacerbate the arthritis despite a good control of liver function. CONCLUSIONS: HCV infection seems to be, possibly in genetically predisposed patients, responsible for arthritis at times similar to rheumatoid arthritis. In these patients a careful assessment of the interferon alpha treatment is mandatory owing to the potential "arthritogenic effect" due to its immunoregulatory properties. PMID- 12563366 TI - [Detection of serum anti-B/B' UsnRNP antibodies in patients with connective tissue diseases by immunoblotting]. AB - OBJECTIVE: To investigate the reliability of the immunoblot method in the detection of serum immunoreactivity towards the B/B' polypeptides of U small nuclear ribonucleoproteins (UsnRNP) and to assess the significance of these antibodies in connective tissue disease (CTD) patients. METHODS: We tested the sera of 348 patients with CTD (101 SLE, 51 systemic sclerosis, 53 primary Sjogren's syndrome, 27 poly/dermatomyositis, 15 rheumatoid arthritis and 101 overlap CTD), of 31 matched healthy subjects and 13 patients with primary Epstein Barr virus (EBV) infection with high titre of IgG anti-EBV antibodies. IgG anti UsnRNP antibodies were determined by immunoblotting on nuclear extract from Raji cells (an EBV-immortalised human B lymphoid cell line) and Jurkat cells (a human T lymphoid cell line). Anti-dsDNA antibodies were detected by indirect immunofluorescence on Crithidia luciliae and anti-ENA by counterimmunoelectrophoresis. Anti-dsDNA activity and avidity were measured in SLE sera by ELISA with Scatchard analysis. Results were statistically analysed by chi-square and Mann-Whitney tests. RESULTS: A high frequency of anti-B/B' antibodies was found in the sera of CTD patients, confined to SLE (54.4%) and overlap CTD with SLE features (55,2%). Anti-B/B' immune reactivity was closely associated with other anti-UsnRNP specificities, gel precipitating anti-nRNP and anti-P antibodies. Nine out of 15 (60%) anti-B/B' positive/anti-ENA negative lupus sera on Raji blots were confirmed to be positive also on Jurkat blots. The sera from patients with EBV infection provided, on Raji blots, completely different band patterns from those obtained with auto-immune sera. CONCLUSIONS. The Sm B/B' proteins are the predominant or, at least, the most frequently targeted antigens of the UsnRNP auto-immune response in SLE and "lupus-like" overlap CTD. Moreover, anti-B/B' is diagnostically specific for CTD with SLE features. Immunoblotting on human B lymphoid cells is a reliable method, in terms of sensitivity and specificity, for the detection of anti-Sm B/B' antibodies. PMID- 12563367 TI - [Evaluation model of the effect of Rofecoxib on the co-prescription of gastroprotective agents observed during the treatment of osteoarthritis]. AB - OBJECTIVE: This study was conducted to define an evaluation model to estimate changes in the co-prescription of gastroprotective agents (GPAs) induced by rofecoxib in the treatment of osteoarthritis (OA). METHODS: On the basis of a cross-linking information, which were stored in different administrative and clinical databases, a multivariate regression analysis was used to develop the model. Data were collected by 30 general practitioners of the Local Health Unit of Ravenna (middle-north of Italy). RESULTS: The study population consisted of 2,944 patients treated with non-steroidal anti-inflammatory drugs (NSAIDs) and 487 treated with rofecoxib. Patients treated with rofecoxib generally presented a higher number of gastrointestinal damage risk factors and also a lower level of GPAs co-prescription compared to those treated with NSAIDs. Including in the model variables such as type of anti-inflammatory treatment (NSAIDs or rofecoxib), gender, age by class, previous hospital admissions due to gastrointestinal complications, number of different NSAIDs used, and prescription of corticosteroids, the regression equation and its coefficients were identified. A non-linear relationship between the percentage of patients treated with rofecoxib and the relative reduction of GPAs co-prescription was found. It has been estimated the basis of the registered percentage of patients treated with rofecoxib (17,6%) adjusting for gastrointestinal damage risk factors, and on a 63% (CI95%: 55%-70%) relative reduction of GPA use with rofecoxib with respect to NSAIDs was estimated. CONCLUSIONS: Based on data collected in the clinical practice after the introduction of rofecoxib, a model evaluating the relationship between the frequency of its use in the OA population and the expected reduction of GPAs, has been developed. PMID- 12563368 TI - [Use of nonsteroidal anti-inflammatory drugs in patients with vertebral osteoporotic fractures]. AB - OBJECTIVE: The aim of the study was to assess the use of Non Steroidal Anti Inflammatory Drugs (NSAIDs) in patients with a history of osteoporotic vertebral fractures. METHODS: We investigated 119 patients with postmenopausal osteoporosis complicated by one or more non recent vertebral fractures. RESULTS: More than 60% of the patients took at least one dose of NSAID weekly. The most prescribed NSAID was nimesulide, at a dose with an exclusively antalgic effect. Patients with wedge fracture and those with a documented vertebral fracture in the last 12 months were those taking NSAIDs more frequently. 77% of the patients that used NSAIDs had concomitant features of osteoarthritis, mainly at the spine or at the knee. The use of NSAIDs was negatively related to the use of specific therapy for osteoporosis, particularly for oral daily tablets. CONCLUSIONS: This study highlights the significant use of NSAIDs in patients with osteoporotic vertebral fractures and the overlap between osteoporosis, osteoarthritis and related treatments. PMID- 12563369 TI - [Anti-tissue transglutaminase antibodies in inflammatory and degenerative arthropathies]. AB - Recent studies identified tissue transglutaminase (tTG) as the antigen eliciting antiendomysial antibodies (EMA) in celiac disease (CD). Anti-tTG antibodies have therefore been proposed as a serological test for CD. Nevertheless, IgA anti-tTG but not EMA have also been found in inflammatory bowel disease patients, suggesting that these antibodies are linked to a tissue lesion rather than to an auto-immune component of CD. To confirm this hypothesis, we evaluated the presence of IgA anti-tTG in patients with inflammatory and degenerative diseases, in whom tissue lesions presented far away from the intestinal mucosa. The study was carried out on the serum and synovial fluid (SF) of 68 patients with rheumatoid arthritis (RA=33), psoriatic arthritis (PsA=26) and osteoarthritis (OA=9). In RA, PsA and OA sera, IgA anti-tTG were positive in 33%, 42% and 11% of patients, respectively. Serum anti-tTG levels were significantly higher in RA (p<0.0001), PsA (p<0.0001) and OA (p<0.02) with respect to healthy controls. SF anti-tTG levels were significantly higher in PsA (p<0.018) than in OA. A good correlation between serum and synovial fluid anti-tTG levels was found in all arthropathies This study suggests that tTG is not the only antigen of EMA and, furthermore, that IgA anti-tTG antibodies represent a general lesion-associated event. Moreover, the significant correlation between serum and synovial fluid anti-tTG levels allow us to hypothesise that these antibodies could be synthesized in the site of arthritic lesions. PMID- 12563370 TI - [Autologous stem cell transplantation in a patient with diffuse systemic sclerosis]. AB - Systemic Sclerosis (SSc) is a systemic disease of unknown etiology presenting with disseminated skin thickening and fibrotic impairment of various organs including lung and kidney. According to the rate and degree of skin involvement, SSc can be classified in a limited and a diffuse form, the latter showing a severe and progressive lung involvement, which is responsible for its high related morbidity and mortality along with resistance to standard therapeutic protocols. High dose chemotherapy, followed by autologous stem cell transplantation, is a standard therapeutic regimen for haematological diseases: re-infusion of mobilised peripheral blood progenitor cells overcomes the myeloablative effect of super-maximal eradicative doses of chemotherapeutic agents. Recently, this therapeutic approach has been applied in some cases of resistant SSc and, albeit the low number of cases, it has been proven effective in early diagnosed and rapidly progressive forms of the disease showing a clinical improvement and an instrumentally detectable decrease of fibrosis extent. We report the case of a young woman affected by diffuse SSc with a rapid progression of clinical signs and instrumentally detectable lesions who underwent a conditioning regimen with fludarabine, cyclophosphamide and anti thymoglobulines followed by re-infusion of autologous peripheral blood stem cells. Two years after transplantation a clinical and instrumental evidence of treatment was observed, with good control of disease evolution. The only sign of disease resumption was a slow worsening of skin involvement. PMID- 12563371 TI - [Case report: rheumatoid arthritis and large granular lymphocytes syndrome]. AB - Felty's syndrome (FS) is a rare complication (less than 1%) of rheumatoid arthritis (RA), with the clinical feature of splenomegaly and neutropenia. Approximately 10-40% of FS patients have an expansion of peripheral blood large granular lymphocytes (LGL). This cell population mainly consists of two subsets: cytotoxic T cells (CD8+, CD57+) and natural killer cells (CD3-,CD8-,CD56+). It has been hypothesised that LGL expansion could be responsible for neutropenia by suppressing neutrophil precursors in the bone marrow, but various mechanisms have been proposed to explain this association. We report a case of a 60-year-old woman with rheumatoid factor positive RA who developed LGL expansion responsible for splenomegaly, but without neutropenia. In conclusion, LGL expansion is an uncommon complication of RA and may be responsible for both FS and clinical pictures resembling FS. PMID- 12563372 TI - [Power Doppler e mezzi di contrasto nello studio della membrana sinoviale reumatoide]. AB - Pannus formation is a fundamental event in the pathogenesis of rheumatoid arthritis and its hypervascularisation seems to be crucial to the development of joint damage. High-resolution greyscale ultrasonography is a safe, quick, and inexpensive imaging tool that allows an accurate detection of even minimal morphostructural changes in patients with rheumatoid arthritis, including joint effusion, thickening of synovial membrane and bone erosions. More recently, power Doppler sonography has proved to be a reliable tool for semiquantitative assessment of the vascularity of the synovial tissue. The contrast-enhanced power Doppler sonography seems to be a helpful adjunct in assessing synovitis and the therapeutic response to the different therapies in patients with rheumatoid arthritis. The aim of this radiological vignette was to show a representative example of use of power Doppler sonography with contrast agent in assessing rheumatoid synovitis. PMID- 12563373 TI - [Tissue engineering applications: cartilage lesions repair by the use of autologous chondrocytes]. AB - Promising new therapies based on tissue engineering have been recently developed for cartilage repair. The association of biomaterials with autologous chondrocytes expanded in vitro can represent a useful tool to regenerate this tissue. The scaffolds utilised in such therapeutical applications should provide a pre-formed three-dimensional shape, prevent cells from floating out of the defect, have sufficient mechanical strength, facilitate uniform spread of cells and stimulate the phenotype of transplanted cells. Hyaff-11 is a hyaluronic-acid based biodegradable polymer, that has been shown to provide successful cell carrier for tissue-engineered repair. From our findings we can state that human chondrocytes seeded on Hyaff-11 are able to maintain in vitro the characteristic of differentiated cells, expressing and producing collagen type II and aggrecan which are the main markers of cartilage phenotype, down-regulating collagen type I. Moreover, it seems to be a useful scaffold for cartilage repair both in animal models and clinical trials in humans, favouring the formation of a hyaline-like tissue. In the light of these data, we can hypothesise, for the future, the use of autologous chondrocyte transplantation together with gene therapy as a treatment for rheumatic diseases such as osteoarthritis. PMID- 12563374 TI - ["Gout and the spider" by Jean de La Fontaine (1621-1695), or the metamorphoses of a rheumatology tale]. AB - By starting from the critical analysis of the fable "Gout and the spider"(Fables Choisies mises en vers, III,8) by Jean de La Fontaine, the author inquires into the historical and literary sources of such a tale. They lie in a short story by Francesco Petrarca ("Aranea et Podagra"), that can be found in a letter from the "Epistolae de Rebus Familiaribus" (III,13). Antecedent sources can be recognized in the Exempla by Jacques de Vitry and Jean Gobi, as well as in a brief poem by Paulus Diaconus. PMID- 12563375 TI - Juvenile dermatomyositis: clinical, laboratorial, histological, therapeutical and evolutive parameters of 35 patients. AB - This study was based on a prospective and a retrospective analysis of 35 patients who met Bohan and Peter criteria for juvenile dermatomyositis diagnosis. The mean follow-up time was three years ten months. Calcinosis was present in five (14.28 %) patients, cutaneous ulcers in four (11.42%), and systemic involvement in nine (27.71%) patients. All patients presented alterations in the serum levels of muscle enzymes, and all of them were submitted to muscle biopsy as a diagnostic procedure. Nine (25.71%) patients received corticotherapy prior to and 26 (74.28%) after the muscle biopsy. Chloroquine, methotrexate, cyclosporine, cyclophosphamide and intravenous immunoglobulin were used in patients with poor response to corticotherapy. Continuation of cutaneous manifestations was observed in 4 (11.43%) patients, laboratorial activity in 1 (2.85%), cutaneous and laboratorial activities in 3 (8.57%). Ten (28.57%) patients were out of activity, and 17 (48.57%) in remission at study end-point, on March 2002. Two (5.71%) patients died. PMID- 12563376 TI - The cerebral correlates of set-shifting: an fMRI study of the trail making test. AB - The trail making test (TMT) pertains to a family of tests that tap the ability to alternate between cognitive categories. However, the value of the TMT as a localizing instrument remains elusive. Here we report the results of a functional magnetic resonance imaging (fMRI) study of a verbal adaptation of the TMT (vTMT). The vTMT takes advantage of the set-shifting properties of the TMT and, at the same time, minimizes the visuospatial and visuomotor components of the written TMT. Whole brain BOLD fMRI was performed during the alternating execution of vTMTA and vTMTB in seven normal adults with more than 12 years of formal education. Brain activation related to the set-shifting component of vTMTB was investigated by comparing performance on vTMTB with vTMTA, a simple counting task. There was a marked asymmetry of activation in favor of the left hemisphere, most notably in dorsolateral prefrontal cortex (BA 6 lateral, 44 and 46) and supplementary motor area/cingulate sulcus (BA 6 medial and 32). The intraparietal sulcus (BA 7 and 39) was bilaterally activated. These findings are in line with clinico-anatomic and functional neuroimaging data that point to a critical role of the dorsolateral and medial prefrontal cortices as well as the intraparietal sulci in the regulation of cognitive flexibility, intention, and the covert execution of saccades/anti-saccades. Many commonly used neuropsychological paradigms, such as the Stroop, Wisconsin Card Sorting, and go - no go tasks, share some patterns of cerebral activation with the TMT. PMID- 12563377 TI - Diffusion-weighted sequence on MRI for the diagnosis of Creutzfeldt-Jakob disease. AB - Creutzfeldt-Jakob disease (CJD) is a progressive and fatal dementing illness caused by a virus like agent called prion. Currently, the definitive diagnosis can only be made through brain biopsy. Given its potential transmissibility, it is paramount to have noninvasive and reliable means to detect the disease. The present case reports on a 63 year-old man with biopsy proven CJD, and evaluates the dependability of diffusion-weighted MRI in this condition, stressing the importance of this particular sequence to its diagnosis. PMID- 12563378 TI - Number and viability of parasite influence seizure frequency in children with neurocysticercosis. AB - We investigated the relationship between CT findings - number and the viability phase of the parasites - and the seizure frequency in children with neurocysticercosis before and short-term after antiepileptic drug (AED) introduction. We only found a significant interaction between stage of parasitic infection and number of lesions on seizure frequency after AED treatment. Patients with more than five lesions on active or transitional stages had higher seizure frequency predicting a worse short-term prognosis. PMID- 12563379 TI - Clinical features of episodic migraine and transformed migraine: a comparative study. AB - Transformed migraine (TM) is one of the most frequent types of chronic daily headache. Eighty patients: 40 with episodic migraine (EM) and 40 with TM with ages ranging from 18 to 60 years old were studied. Females were the majority. At first examination, the mean age was similar in both groups. The initial age of migraine attacks was significantly smaller in the TM group. Time history of episodic attacks was similar in both groups. In the EM group, the headache was predominantly located on only one side of the head; whereas in the TM group, on more than one side. There was variation in the character of pain and intensity in the TM group. Nocturnal awakening with headache, aura and family history did not show significant association with EM or TM. The TM was distinguished from the EM in relation to the frequency, location and pain intensity of the headache. Patients with early migraine headache onset may exhibit a further risk of developing TM. PMID- 12563380 TI - Progressive supranuclear palsy in a sample of Brazilian population: clinical features of 16 patients. AB - Progressive supranuclear palsy (PSP) is an uncommon disorder characterized by marked postural instability, vertical gaze abnormalities and axial rigidity. The purpose of this study is to report the clinical features of 16 consecutive subjects seen over a 10-year period at a Movement Disorders Clinic. These subjects fulfilled criteria for probable PSP namely those of the National Institute of Neurologic Disorders and Stroke (NINDS) and the Society for PSP (SPSP). This patient-group represented 2.1% of all degenerative parkinsonians observed and the mean age of onset of the disease was 64.7 years (sd = +/- 7.2). Postural instability with falls was the most frequent initial feature presented in PSP patients (62.5%). The hallmark of the disease, the supranuclear vertical gaze palsy, appeared after 2.3 years of disease onset, and only 12.5% had such manifestation at the first evaluation. Transient tremor was observed with a relatively high frequency in this group (44%), but only 19% had rest tremor. Chronic dacryocystitis, probably related to a paucity of blinking, was observed in two patients as an inaugural manifestation. In the first evaluation, only 19% of the 16 patients were diagnosed as probable PSP. The mean interval prior to the final diagnosis was 2.4 years. PMID- 12563381 TI - Usefulness of additional nerve conduction techniques in mild carpal tunnel syndrome. AB - This study was done to assess the percentage of abnormality in additional nerve conduction techniques after normal median distal latency (routine) in mild carpal tunnel syndrome (CTS). Bilateral nerve conduction studies were carried out in 116 consecutive symptomatic CTS patients (153 hands). Mild cases were based on normal routine (< 3.7 ms, peak-measured, 14 cm) and at least one technique abnormal of the following: sensory median-radial difference (MR); sensory median-ulnar difference (MU4); mixed palm median-ulnar difference (MUP); median palm latency (PW); and motor median distal latency (MDL). After normal cut-off values for routine, 3.1 to 3.6 ms (< 3.7 ms), we found an abnormal MR, ranging from 86.6 to 93.4%, followed by MU4 (40 to 81.7%), MUP (20 to 71.2%), PW (0 to 41.1%), and MDL (0 to 19.6%). The most frequent abnormal association were MR plus MU4 in 90.1%, followed by MR plus MUP and MU4 plus MUP. The most frequent abnormal additional nerve conduction technique for mild CTS electrodiagnosis was MR, followed by MU4 and MUP. Percentage of MR abnormality was very high regardless of the median routine latency cut-off (< 3.1 to < 3.6 ms). PMID- 12563382 TI - Neonatal treatment with fluoxetine reduces depressive behavior induced by forced swim in adult rats. AB - Serotonin plays a role at the pathophysiology of depression in humans and in experimental models. The present study investigated the depressive behavior and the weigh evolution in adult rats (60 days) treated from the 1st to the 21st postnatal day with fluoxetine, a selective serotonin reuptake inhibitor (10 mg/kg, sc, daily). The depressive behavior was induced by the forced swim test (FST). The animals were submitted to two sessions of FST: 1st session for 15 min and the 2nd session 24h later, for 5 min. During the 2nd session the Latency of the Attempt of Escape (LAE) and Behavioral Immobility (BI) were appraised. The Fluoxetine group when compared to the Control group, showed an increase in LAE and a decrease in BI. The neonatal administration of fluoxetine reduced the depressive behavior in adult rats, possibly by increase in the brain serotonergic activity. This alteration can be associated to process of neuroadaptation. PMID- 12563383 TI - Third ventriculostomy through the lamina terminalis for intracranial pressure monitoring after aneurysm surgery: technical note. AB - OBJECTIVE: A new ventriculostomy technique through the lamina terminalis is described. This technique is applied mainly during aneurysm surgery at the acute stage. METHOD: Thirteen patients were operated on intracranial aneurysms and, during the procedure, had the lamina terminalis fenestrated. A ventricular catheter was inserted into the third ventricule, left in place and connected to an external drainage system for further intracranial pressure (ICP) monitoring and/or cerebrospinal fluid (CSF) drainage. RESULTS: ICP readings and CSF drainage were obtained in all cases. No complication was recorded. CONCLUSION: Third ventriculostomy through the lamina terminalis is a simple and easy technique that can be used as an alternative to conventional ventriculostomy. This procedure can be indicated in cases where the ventricule is not reached by means of another technique, and when the decision to perform ventriculostomy is made at the end of aneurysm surgery. PMID- 12563384 TI - Unilateral subthalamic nucleus lesioning: a safe and effective treatment for Parkinson's disease. AB - The present study, the largest in the literature, was performed to assess the effectiveness and safety of unilateral subthalamic nucleus (STN) lesioning for Parkinson's disease (PD). From August 1999 to September 2000, 21 consecutive patients evaluated pre- and postoperatively by a single examiner were operated. Levodopa intake and dyskinesia, Hoehn & Yahr, Schwab & England and UPDRS motor scores were recorded. Stereotactic CT and MRI and the effects of macrostimulation were used to determine STN coordinates. A single radiofrequency lesion was made (60-75 degrees C/60"). Concomitant ipsilateral Vim/VOp lesions were made in 8 patients. Using a new technique, we were able to determine the territory of STN involved by the surgical lesion. The Wilcoxon and Mann-Whitney statistical tests were applied to evaluate the surgical results. All recorded parameters showed stable improvement after a mean follow up of 13.5 months. Recurrence occurred in two patients. Contralateral tremor arrest and decrease of rigidity and bradykinesia should be regarded as STN hallmarks to stimulation. Hyperintense lesions in the early-phase MRI seem to be a poor prognostic factor. Lateral territory lesioning correlates with better results. There was no significant difference between the cohorts with and without a Vim/VOp lesion. Dyskinesias happened in two patients (promptly abolished by a Vim/VOp lesion). Other complications were transient and/or rare. In conclusion, STN lesioning is a safe and very effective procedure to treat PD and probably an underutilized operation for those who can not afford the costs of DBS. PMID- 12563385 TI - Structural and functional analyses of the occipital cortex in visual impaired patients with visual loss before 14 years old. AB - Single photon emission tomography (SPECT) perfusion images of the brain of individuals with complete visual loss before the age of 14 were carried out and compared to those of visually normal subjects, in order to assess hypothetical differences in brain structural and metabolism between the two groups. Study group was comprised by 2 females and 3 males, aged 30 +/- 10 years. Controls were composed by 6 females and 5 males aged 41.5 +/- 3.8 years. All individuals were submitted to physical and neurological examinations, and to MRI and to SPECT. Blind subjects presented larger perfusion measurements bilaterally in their medial temporal lobes (p=0.030, right side; p=0.01, left side), but smaller perfusion measurements in their left frontotemporal area than controls (p=0.026). Intragroup analysis of the study group disclosed asymmetric perfusion, lesser in the left temporal and parietal areas (p=0.026 and p<0.0001, respectively) compared to the right side. In the healthy controls, reduced perfusion was also noted at the left parietal areas when compared to the right side (p=0.035). The study revealed that completely blind patients that became visually impaired before the age of 14 in spite of not having MRI detectable changes in their brain's anatomy do present increases in perfusion of their left and right medial temporal lobes, and a reduction in the perfusion of the left frontotemporal area, as compared to normal controls. While the increases in blood flow may reflect compensatory mechanisms for visual deprivation, the significance of the diminished perfusion in the left frontotemporal area remains elusive. PMID- 12563386 TI - Developmental study of early posture control in preterm and fullterm infants. AB - In a prospective study, 10 infants born between 32 to 36 weeks of gestation and 10 infants born at fullterm were observed on day 15, and in months 1, 2 and 3. The investigation showed that the development of early postural control takes place in a sequential way in preterm infants, similar to that of in fullterm infants. However, some movement components for the acquisition of motor abilities showed a different trend in the development of preterm infants when compared to fullterm infants: the onset for the acquisition of the extensor and flexor patterns was slower and the distribution of the load bearing was less mature. PMID- 12563387 TI - Cerebrovascular disease in pediatric patients. AB - Although rare in childhood, stroke may have a serious impact when it happens in this stage of life. Also, it may be the first sign of a systemic disease. We report 12 cases of patients with stroke treated in the Neuropediatrics Unit of Hospital de Clinicas de Porto Alegre (HCPA) from March 1997 to March 2000. All patients, from term infants to 12-year-old children hospitalized in the Pediatrics Unit of HCPA, had clinical suspicion of stroke, which was later confirmed by radiological studies. Patient follow up ranged from 1 to 6 years (mean = 3.4 years). Presenting symptoms were hemiparesis in 9 patients, seizures in 7, deviation of labial commissure in 3, and loss of consciousness in 1. The increase in the number of cases of childhood stroke identified and later confirmed by noninvasive methods had helped in the determination of different ethiologies of stroke: the most frequent being hematologic, cardiac and genetic diseases. However, our study included 6 newborns with stroke whose ethiology was not identified. Seven children with seizures received phenobarbital. Six term infants had neonatal seizures secondary to stroke and restricted to the first 72 hours of life. PMID- 12563388 TI - Parachute and lateral propping reactions in preterm children. AB - A non-controlled, prognostic cohort study was performed with the aim of establishing markers of neurological development and defining a clinical and epidemiological profile of preterm newborns at 3, 6, 9, and 12 months of gestation-corrected age in terms of parachute and lateral propping reactions. Newborns with gestational age of up to 36 weeks and 6 days, weighing 2,000 g or less at birth, were included in the study At 6 months of age, parachute and lateral propping reactions were present in 8.1% of the patients. At 9 months, the parachute reaction was present in 87.5%, and the lateral propping reaction was present in 90% of the children. It was possible to assess parachute and lateral propping reactions in preterm children in the first year of life. Alterations in trunk-limb coordination may be evidenced in the 1st year of life through postural reactions, which are maintained as prematurity markers until school age. PMID- 12563389 TI - [Dementia in patients of Hospital das Clinicas da UNICAMP]. AB - OBJECTIVE: To report the relative frequencies of the causes of dementia in patients seen at UNICAMP Hospital. METHOD: From 1989 to 1998, 261 dementia patients were admitted (89.7% studied retrospectively, and 10.3%, prospectively); mean age 63.5 years (+/-13.2), and education 3.6 years +/- 3.9; 25% illiterate). Diagnosis of dementia and its subtypes was based on DSM-IV criteria, CAMDEX, NINCDS-ADRDA, NINDS AIREN/ADDTC, Lund-Manchester, besides tap-test, including laboratory and neuroimaging examinations. RESULTS: Sixty five patients (24.9%) had vascular dementia, 62 ( 23.7%) Alzheimer's, 29 (11.1%) depressive pseudodementia, 28 (10.9%) normal pressure hydrocephalus, 14(5.4%) mixed, 12 (4.6%) post-traumatic, 9 (3.4%) frontotemporal and 38 (14.5%) had diverse or unknown causes. Dementia was minimal or mild in 123 (47.1%), moderate in 95 (36.4%), and severe in 43 (16.5%). CONCLUSION: Frequency of vascular dementia was 24.9%, Alzheimer's 23.7%, and normal pressure hydrocephalus 10.7%, in disagreement with that found in the neurological literature (20-30%, 50%, and 1 4%, respectively), probably because we are dealing with cases admitted to a tertiary university hospital. Epidemiological studies are needed to disclose the true proportion of these dementia syndromes in the population. PMID- 12563390 TI - [Analysis of 52 patients with head trauma assisted at pediatric Intensive Care Unit: considerations about intracranial pressure monitoring]. AB - OBJECTIVES: Analysis of 52 pediatric patients with head trauma assisted at Intensive Care Unit; to present considerations about epidemiologic factors of trauma, clinical presentation, tomografic aspects, hemodynamic changes and treatment options of intracranial hypertension; to present considerations about the intracranial pressure (ICP) monitoring. METHOD: Retrospective study involving 52 patients with head trauma and 17 patients submitted to a ICP monitoring. RESULTS: We found a male predominance, mean age 7.75 years-old, main cause was run over (38.5%); 21.2% patients presentd arterial hypotension; 67.3% were considered severe head trauma. According to Marshall tomografic grading we had 19.2% type I, 65.4% type II, 3.8% type III, 3.8% type IV and 7.7% type V. Seizures occurred in 25% children ICP monitoring was made in 32.7% of all patients. Mortality rate was 11.5%. In 58% the maximum ICP level occured at the second day of trauma. CONCLUSION: Prognosis was related to severity of trauma, arterial hypotension, Marshall's tomografic gradind III and IV and ICP high values. The ICP monitoring was considered useful to allow the identification and treatment of intracranial hypertension. PMID- 12563391 TI - [The baby retraction reaction evaluation in pediatric diseases: a validity study]. AB - The evaluation of the withdrawal reaction in small children using a standardized instrument should take into account the child's psychopatologic context and its child's development moment. We present the BADS questionnaire validation by its application in 35 children aged 0 to 2 years-old with pediatric diseases, coming from Santa Casa de Sao Paulo. The results were compared with those of 90 normal children who were evaluated in a previous study of the same author. The test used was the independent t test, with mean scores of 5.90 +/- 2.57 for normal children and 6.37 +/- 4.83 for sick children, with p= 0,651 and significance level of 5%. Thus, it was observed that the withdrawal reaction in sick children is not significantly different for that for normal children. With these results, the scale shows its importance as a screening instrument. PMID- 12563392 TI - [Study of motor function development at 8 and 12 months of age in preterm and at term children]. AB - OBJECTIVE: To compare the development of motor function in children born preterm with those born at term, at 8 and 12 months of age. To investigate the relation of motor function quality at the age of 8 months with motor ability at 12 months. METHOD: Thirty-two children participated in this study: 16 were born preterm (risk group) and 16 were born at term (control group). The spontaneous movements of the children were assessed at 8 months and their mobility skills and independence were assessed at 12 months (corrected ages for the preterm group), using standardized developmental tests (AIMS and PEDI, respectively). Data were analysed using independent t-tests (between-group comparison) and Pearson correlation coefficients (within-group comparison). RESULTS: There was no significant difference in motor function, between those born preterm with those born at term, either at 8 or at 12 months of age. In the control group, there was significant association (r=0.67; p=0.004) between movement at 8 months and mobility skills at 12 months. In the risk group, there was significant relationship between skills and independence in mobility, at 12 months corrected age (r=0.80; p=0.0001). CONCLUSION: Preterm born children, without other disorders and with age correction, might show a similar motor development as those born at term. The path for the acquisition of motor abilities in preterm born children appears to differ among those infants. PMID- 12563393 TI - [Characteristics of the communication in individuals with fragile X syndrome]. AB - The aim of this study was to make a characterization of the communication in a group with fragile X syndrome (FXS), based on normal development scale. The sample has 10 boys, between 6 and 13 years and with FXS. All of them were submitted to language evaluation. The most part of them had the communication in linguistic form (80%), until 3 years old. It was considered that the data confirmed the significant delay in the acquisition and development of the communication, being necessary an early and specialized intervention. PMID- 12563394 TI - [Clinical, endoscopical and manometric evaluation of swallowing in patients with acquired autoimmune myasthenia gravis]. AB - Dysphagia, or difficulty swallowing, is a common problem following myasthenia gravis (MG) and may lead to aspiration of saliva, food or liquids. We herein present 22 MG patients, with complaint of dysphagia, evaluated by phonoaudiological evaluation, nasofibrolaryngoscopical analysis and manometry of upper esophageal sphincter. The main objective was to evaluate the phases of the swallowing process and anatomical and functional aspects of oropharyngeal musculature. The age of patients varied from 19 to 74 years; being 19 female and 3 male. The main data were: (1) statistically significant relation between the phonoaudiological clinical evaluation and nasofibrolaryngoscopical one; (2) stomatognatical system disorders present in 100%; (3) swallowing and chewing disorders present in 100%, when clinically evaluated, and in 81,8% when evaluated by the nasofibrolaryngoscopy; (4) statistically significant relation between penetration/aspiration and antecedents of pneumonia; (5) stomatognatical muscles very altered, difficulty controlling the bolus, collection of food in the pharynx or larynx and presence of penetration and/or aspiration showed significant correlation with weakness of a pharyngeal muscles. These findings justify the necessity to evaluate clinically the swallowing phases in MG patients, with the main purpose to prevent both aspiration and dehidration from inadequate oral intake. PMID- 12563395 TI - Pathological laughter in a patient with trigeminal neurinoma. AB - We present a 47-year-old woman with a long history of anxiety and a more recent history of shock-like facial pain and episodes of laughter without any motivation. She could not explain the laughing bursts and did not have a sense of mirth preceding it. On neurological examination she presented a VI nerve palsy and trigeminal hypoesthesia (V2 and V3) on the right side. Magnetic resonance imaging exhibited a large cystic lesion on the right middle fossa causing significant compression on the brain stem. A frontoorbitozygomatic and pretemporal combined approach was performed. During intra and extradural exploration a large tumor was found on the trigeminal nerve. The whole lesion was resected, revealing to be a neurinoma on pathological exhamination. She maintained a VI nerve palsy but had complete remission of the unmotivated laughing episodes during the one year follow up. PMID- 12563396 TI - Organophosphate-induced delayed neuropathy: case report. AB - Organophosphate induced delayed neuropathy (OPIDN) is an uncommon clinical condition. It occurs in association with the ingestion of great amounts of organophosphate after the stimulation of cholinergic receptor. The clinical picture is characterized by a distal paresis in lower limbs associated with sensitive symptoms. Electrodiagnostic studies show a motor axonal neuropathy. Involvement of the central nervous system may occur. We describe a 39 years-old female patient who developed hyperesthesia associated with lower limbs paresis, fourteen days after she had ingested a Dichlorvos-based insecticide. Electrophysiological study was characterized by an axonal polyneuropathy pattern. Pyramidal tract dysfunction was observed later in upper limbs. Considering that both peripheral and central nervous systems are involved we believe that the more appropriated term would be organophosphate induced delayed neuropathy (OPIDN) instead of organophosphate induced delayed polyneuropathy (OPIDP). PMID- 12563397 TI - Choroid plexus papilloma and Aicardi syndrome: case report. AB - A case of Aicardi syndrome associated with a choroid plexus papilloma of the third and both lateral ventricles in a 15 months old baby girl is repported. The tumor was completely removed via three craniotomies. Reports of the literature with the association of choroid plexus papilloma and Aicardi syndrome are rare. We suggest that children diagnosed with Aicardi syndrome should routinely undergo imaging studies of the brain, such as computed tomography or magnetic resonance. PMID- 12563398 TI - A further case of a Prader-Willi syndrome phenotype in a patient with Angelman syndrome molecular defect. AB - Angelman syndrome (AS) and Prader-Willi syndrome (PWS) are distinct human neurogenetic disorders; however, a clinical overlap between AS and PWS has been identified. We report on a further case of a patient showing the PWS phenotype with the AS molecular defect. Despite the PWS phenotype, the DNA methylation analysis of SNRPN revealed an AS pattern. Cytogenetic and FISH analysis showed normal chromosomes 15 and microsatellite analysis showed heterozygous loci inside and outside the 15q11-13 region. The presence of these atypical cases could be more frequent than previously expected and we reinforce that the DNA methylation analysis is important for the correct diagnosis of severe mental deficiency, congenital hypotonia and obesity. PMID- 12563399 TI - [Paracoccidioidomycosis meningitis: case report]. AB - The involvement of the central nervous system in paracoccidioidomycosis and its meningeal form of clinical presentation have rarely been described. This is a case report of a 23 years old man who developed paracoccidioidomycosis meningitis achieved by fungus' presence in direct research and culture of cerebrospinal fluid. The meningeal form of paracoccidioidomycosis, its laboratorial and imaging diagnostic are discussed, and the importance of the routine investigation of the fungus is emphasized. PMID- 12563400 TI - [Aneurysm of the anterior inferior cerebellar artery: case report]. AB - The intracranial aneurysms of the posterior circulation have been reported between 5 and 10% of all cerebral aneurysms and the aneurysms of the anterior inferior cerebellar artery (AICA) are considered rare, can cause cerebello pontine angle (CPA) syndrome with or without subarachnoid hemorrhage. Since 1948 few cases were described in the literature. We report on a 33 year-old female patient with subarachnoid hemorrhage due to sacular aneurysm of the left AICA. She was submitted to clipage of the aneurysm without complications. PMID- 12563401 TI - [Worsening of autosomal dominant restless legs syndrome after use of mirtazapine: case report]. AB - We report the case of a 78 years old female patient with primary restless legs syndrome (RLS) with an autosomal dominant pattern of inheritance. In addition, the patient also had depression. We emphasize the worsening of symptoms of RLS after the use of a selective serotonin uptake inhibitor (mirtazapine), with improvement after the drug was discontinued, and an excellent recovery with the use of low dose dopaminergic agonist (pramipexol). PMID- 12563402 TI - [Central pontine and extra-pontine myelinolysis in an alcoholic patient without hydro-electrolyte disturbances: case report]. AB - Central pontine myelinolysis (CPM) and extra-pontine myelinolysis (EPM) are different presentations of a demyelinating disorder of the brain more commonly associated with rapid correction of hyponatremia, spastic tetraparesia and pseudo bulbar palsy. There are in the literature a few cases of CPM/EPM in patients without electrolyte disturbances. We report the case of a 39 year-old man with severe alcoholism, who presented with spastic tetraparesis and palsy of several cranial nerves, associated with lesions in the magnetic resonance compatible with CPM/EPM. The patient had a good follow-up after pulse therapy with corticosteroids. PMID- 12563403 TI - [Spontaneous cervical spondylodiscitis caused by Salmonella typhi in an immunocompetent patient]. AB - We report a case of spontaneous cervical spondylodiscitis caused by Salmonella typhi. A 52-year-old man presented in the neurosurgical service with complaints of pain in the cervical and scapular region. Cervical inflammatory disease was suggested by bone scintigraphy and magnetic resonance imaging. The diagnosis of Salmonella typhi spondylodiscitis was established by blood culture and culture of needle biopsy specimen taken from the C5 vertebra. The agglutinin titers for Salmonella were elevated. Intravenous ciprofloxacin therapy and external immobilization with a halo vest were instituted. A review of literature was performed evaluating the clinical, diagnostic and therapeutic aspects of this unusual pathology. PMID- 12563404 TI - [Occipital condyle fracture associated with Jefferson's fracture and injury of lower cranial nerves: case report]. AB - Occipital condyle fracture(OCF) is rarely seen and can be missed during medical evaluation due to the variety of clinical presentations and the difficulty to be visualized radiographically. This fracture can be associated with cranial nerves injuries (31%), being the hipoglossal nerve the most frequently involved (67%). We report a 58 years old female patient who presented with OCF, injury of lower cranial nerves and Jefferson's fracture. The patient was treated with cervical traction for six weeks followed by halo immobilization for three months. There was bone consolidation recovery of the nervous injury after this period. This report emphazises the importance of investigating the skull-cervical transition in all patients with cervical trauma. Although Jefferson's fracture is rarely associated with OCF, it should be remembered and treated appropriately when diagnosed. PMID- 12563405 TI - Gamma probe-assisted brain tumor microsurgical resection: a new technique. AB - OBJECTIVES: The pioneering performance of gamma probe-assisted surgery (GPAS) for brain tumors, aiming not only an improvement of tumor detection, but mainly assurance of its complete removal and the study of the usual distribution of the 99mTc-MIBI in the brain SPECT of normal individuals. METHOD: Patient's informed consent and demonstration of the tumor by the preoperative MIBI SPECT were the inclusion criteria adopted for GPAS, which was performed in one patient with a right parietal lobe metastatic tumor. The radiotracer (99mTc-MIBI) was injected in a peripheral vein 5 hours before the operation. A tumor to-normal tissue count ratio equal to or greater than 2/1 was considered indicative of tumor. MIBI SPECT was performed in five normal individuals in a pilot study. RESULTS: The gamma probe greatly facilitated intraoperative tumor detection (tumor to-normal brain count ratio was 5/1) and indicated a small piece of residual tumor after what was thought to be a complete tumor removal, allowing its resection, which, otherwise, would have been left behind. Postoperative CT confirmed complete tumor resection. The MIBI SPECT in normal individuals showed an increased uptake by the hypophisis, choroid plexus, skull, scalp and salivary glands and absence of uptake by the normal brain tissue. There were no complications. CONCLUSION: GPAS proved to be, in this single case, a safe and reliable technique to improve brain tumor detection and to confirm the presence or absence of residual tumor. PMID- 12563413 TI - A yeast-based model system for cloning secreted and membrane proteins. AB - The targeting of proteins to cell organelles and membranes, or of proteins destined to secretion, is coordinated by signal sequences located at the 5'-end of their respective genes. A signal sequence trap system was envisaged in which a truncated version of the yeast acid phosphatase pho5 gene lacking the start codon and signal sequence could serve as a reporter gene. A fraction enriched in 5'-end fragments obtained by PCR from a potato guard-cell cDNA library was cloned in frame to the acid phosphatase gene and the acid phosphatase activity was assayed directly in yeast colonies grown on selective medium. Putative signal sequences targeting the acid phosphatase to the membrane or to the outside of the cell were used to screen the cDNA bank in order to recover the original full-size sequence which gave rise to the signal sequence. Two unknown sequences displaying marked tissue-specific expression were retrieved, one of them (YE139) with a higher expression level in green buds and stem cells, and the other one (YE290) with a higher expression level in androceum, gyneceum, and roots. The limitations of the system are further analyzed using other sequences as control. PMID- 12563414 TI - A second note on the geographical differentiation of Amphisbaena fuliginosa L., 1758 (Squamata, Amphisbaenidae), with a consideration of the forest refuge model of speciation. AB - The geographical differentiation of Amphisbaena fuliginosa L. is studied with basis on 220 specimens from 99 definite and 2 generic localities, ranging from Panam to Puno in Peru and Goi s in Brasil. The pattern found is considered to support Vanzolini's 1951 scheme of five subspecies, defined by color pattern and by four meristic characters, and differentiated in consequence of Quaternary paleoecological events. Recent developments relating to models of differentiation in Quaternary tropical South America are briefly considered. PMID- 12563415 TI - The surface charge of trypanosomatids. AB - The surface charge of trypanosomatids was evaluated by means of the binding of cationic particles, as visualized by electron microscopy and by direct measurements of the electrophoretic mobility of cells. The results obtained indicate that most of the trypanosomatids exhibit a negatively charged surface whose value is species specific and varies according to the developmental stages. Sialic acids associated with glycoproteins, glycolipids and phosphate groups are the major components responsible for the net negative surface charge of the trypanosomatids. PMID- 12563416 TI - Removal of the cortical projections alters expression of NOS in the different cell types of the superficial layers of the superior colliculus in rats. AB - Nitric oxide has several biological roles and nitric oxide synthase (NOS) is expressed in the nervous system, and co-localizes with NADPH-diaphorase. The superficial layers of the superior colliculus (SC), which receive retinal and cortical inputs, present NADPH-d staining in a sub-population of neurons that include all cell types. We have previously shown, by NADPH-diaphorase, that eye enucleation alters the intracellular distribution of NOS. Here, we studied the effect of cortical ablation on NOS expression by neurons in collicular superficial layers. Our results show that cortical ablation alters the proportion of different NOS-positive cell types, but not the intracellular distribution of the enzyme. PMID- 12563417 TI - Role of laminin bioavailability in the astroglial permissivity for neuritic outgrowth. AB - The mechanisms involved in the failure of an adult brain to regenerate post lesion remain poorly understood. The reactive gliosis which occurs after an injury to the CNS and leads to the glial scar has been considered as one of the major impediments to neurite outgrowth and axonal regeneration. A glial scar consists mainly of reactive, hypertrophic astrocytes. These reactive cells acquire new properties, leading to A non-permissive support for neurons. Astrogial reactivity is mainly characteriized by a high overexpression of the major component of the gliofilaments, the glial fibrillary acidic protein (GFAP). This GFAP overexpression is related to the astroglial morphological response to injury. We hypothesized that modulation of GFAP synthesis, reversing the hypertrophic phenotype, might also reverse the blockage of neuritic outgrowth observed after a lesion. In this article, we review findings of our group, confirming our hypothesis in a model of lesioned neuron-astrocyte cocultures. We demonstrate that permissivity for neuritic outgrowth is related to phenotypic changes induced in reactive astrocytes transfected by antisense GFAP-mRNA. We also found that this permissivity was related to a neuron-regulated extracellular laminin bioavailability. PMID- 12563418 TI - Modulators of axonal growth and guidance at the brain midline with special reference to glial heparan sulfate proteoglycans. AB - Bilaterally symmetric organisms need to exchange information between the left and right sides of their bodies to integrate sensory input and to coordinate motor control. Thus, an important choice point for developing axons is the Central Nervous System (CNS) midline. Crossing of this choice point is influenced by highly conserved, soluble or membrane-bound molecules such as the L1 subfamily, laminin, netrins, slits, semaphorins, Eph-receptors and ephrins, etc. Furthermore, there is much circumstantial evidence for a role of proteoglycans (PGs) or their glycosaminoglycan (GAG) moieties on axonal growth and guidance, most of which was derived from simplified models. A model of intermediate complexity is that of cocultures of young neurons and astroglial carpets (confluent cultures) obtained from medial and lateral sectors of the embryonic rodent midbrain soon after formation of its commissures. Neurite production in these cocultures reveals that, irrespective of the previous location of neurons in the midbrain, medial astrocytes exerted an inhibitory or non-permissive effect on neuritic growth that was correlated to a higher content of both heparan and chondroitin sulfates (HS and CS). Treatment with GAG lyases shows minor effects of CS and discloses a major inhibitory or non-permissive role for HS. The results are discussed in terms of available knowledge on the binding of HSPGs to interative proteins and underscore the importance of understanding glial polysaccharide arrays in addition to its protein complement for a better understanding of neuron-glial interactions. PMID- 12563421 TI - Insights on diagnosis, prognosis and screening of renal cell carcinoma. PMID- 12563422 TI - Incidental and symptomatic renal tumors: impact on patient survival. AB - CONTEXT: Renal cell carcinoma is the third most frequent genitourinary neoplasia, and there is currently an increase in the incidental diagnosis of tumors confined to the kidneys. OBJECTIVE: To study the survival of patients with incidental and symptomatic renal tumors who have undergone nephrectomy. DESIGN: Retrospective. SETTING: Hospital Sirio Libanes and Hospital Beneficencia Portuguesa de Sao Paulo. PARTICIPANTS: 115 patients with diagnosis of renal cell carcinoma, operated on by the same group of surgeons and evaluated by a single pathologist. MAIN MEASUREMENTS: Sex, age and diagnosis method, analyzed in two groups, according to the tumor diagnosis: Group 1 with incidental diagnosis and Group 2 with symptomatic tumors. The anatomopathological characteristics and patient survival in both groups were evaluated. A statistical analysis was performed using the Student t, chi-squared, log rank and Kaplan-Meyer tests. RESULTS: Among the studied patients, 59(51%) had an incidental diagnosis, with 78% diagnosed by ultrasonography, 20% by computerized tomography scan and 2% during surgeries; 56 patients (49%) were symptomatic. Tumor locations were equally distributed between the two kidneys, and the surgery was conservative for 24% of the incidental and 9% of the symptomatic group. In the incidental group only one patient had tumor progression and there was no death, while in the symptomatic group there were 5 progressions and 10 deaths. The 5-year specific cancer-free survival was 100% in the incidental and 80% in the symptomatic group (p = 0.001) while the disease free rate was 98% in the incidental and 62% in the symptomatic group (p < 0001). CONCLUSION: Incidental renal tumor diagnosis offers better prognosis, providing longer disease-free survival. PMID- 12563423 TI - Descriptive study of 192 adults with speech and language disturbances. AB - CONTEXT: Aphasia is a very disabling condition caused by neurological diseases. In Brazil, we have little data on the profile of aphasics treated in rehabilitation centers. OBJECTIVE: To present a descriptive study of 192 patients, providing a reference sample of speech and language disturbances among Brazilians. DESIGN: Retrospective study. SETTING: Speech Pathology Unit linked to the Neurology Division of the Hospital das Clinicas of the Faculdade de Medicina da Universidade de Sao Paulo. SAMPLE: All patients (192) referred to our Speech Pathology service from 1995 to 2000. PROCEDURES: We collected data relating to demographic variables, etiology, language evaluation (functional evaluation, Boston Diagnostic Aphasia Examination, Boston Naming and Token Test), and neuroimaging studies. MAIN MEASUREMENTS: The results obtained in language tests and the clinical and neuroimaging data were organized and classified. Seventy aphasics were chosen for constructing a profile. Fourteen subjects with left single-lobe dysfunction were analyzed in detail. Seventeen aphasics were compared with 17 normal subjects, all performing the Token Test. RESULTS: One hundred subjects (52%) were men and 92 (48%) women. Their education varied from 0 to 16 years (average: 6.5; standard deviation: 4.53). We identified the lesion sites in 104 patients: 89% in the left hemisphere and 58% due to stroke. The incidence of aphasia was 70%; dysarthria and apraxia, 6%; functional alterations in communication, 17%; and 7% were normal. Statistically significant differences appeared when comparing the subgroup to controls in the Token Test. CONCLUSIONS: We believe that this sample contributes to a better understanding of neurological patients with speech and language disturbances and may be useful as a reference for health professionals involved in the rehabilitation of such disorders. PMID- 12563424 TI - Addition of exogenous cytokines in mixed lymphocyte culture for selecting related donors for bone marrow transplantation. AB - CONTEXT: Mixed lymphocyte culturing has led to conflicting opinions regarding the selection of donors for bone marrow transplantation. The association between a positive mixed lymphocyte culture and the development of graft-versus-host disease (GVHD) is unclear. The use of exogenous cytokines in mixed lymphocyte cultures could be an alternative for increasing the sensitivity of culture tests. OBJECTIVE: To increase the sensitivity of mixed lymphocyte cultures between donor and recipient human leukocyte antigen (HLA) identical siblings, using exogenous cytokines, in order to predict post-transplantation GVHD and/or rejection. TYPE OF STUDY: Prospective study. SETTING: Bone Marrow Transplantation Unit, Universidade Estadual de Campinas. PARTICIPANTS: Seventeen patients with hematological malignancies and their respective donors selected for bone marrow transplantation procedures. PROCEDURES: Standard and modified mixed lymphocyte culturing by cytokine supplementation was carried out using donor and recipient cells typed for HLA. MAIN MEASUREMENTS: Autologous and allogenic responses in mixed lymphocyte cultures after the addition of IL-4 or IL-2. RESULTS: In comparison with the standard method, average responses in the modified mixed lymphocyte cultures increased by a factor of 2.0 using IL-4 (p < 0.001) and 6.4 using IL-2 (p < 0.001), for autologous donor culture responses. For donor-versus recipient culture responses, the increase was by a factor of 1.9 using IL-4 (p < 0.001) and 4.1 using IL-2 (p < 0.001). For donor-versus-unrelated culture responses, no significant increase was observed using IL-4, and a mean response inhibition of 20% was observed using IL-2 (p < 0.001). Neither of the cytokines produced a significant difference in the unrelated control versus recipient cell responses. CONCLUSION: IL-4 supplementation was the best for increasing the mixed lymphocyte culture sensitivity. However, IL-4 also increased autologous responses, albeit less intensively than IL-2. Thus, with this loss of specificity we believe that it is not worth modifying the traditional mixed lymphocyte culture method, even with IL-4 addition. PMID- 12563425 TI - Partial HELLP Syndrome: maternal and perinatal outcome. AB - CONTEXT: HELLP syndrome is a severe complication of pregnancy characterized by hemolysis, elevated liver enzymes and low platelet count. Some pregnant women develop just one or two of the characteristics of this syndrome, which is termed Partial HELLP Syndrome (PHS). OBJECTIVE: The objective of this study was to evaluate the repercussions on maternal and perinatal outcomes among women that developed PHS and to compare these women with those whose gestational hypertension or preeclampsia did not show alterations for HELLP syndrome in laboratory tests. DESIGN: Observational, retrospective and analytical study. SETTING: Maternity Department of Hospital das Cl nicas, Faculdade de Medicina de Botucatu, Universidade Estadual Paulista, Botucatu, S o Paulo, Brazil. SAMPLE: Pregnant or post-delivery women who had a blood pressure elevation that was first detected after mid-pregnancy, with or without proteinuria, between January 1990 and December 1995. MAIN MEASUREMENTS: Analysis was made of maternal age, race, parity, hypertension classification, gestational age at the PHS diagnosis, alterations in laboratory tests for HELLP syndrome, time elapsed to discharge from hospital, maternal complications, mode of delivery, incidence of preterm birth, intrauterine growth restriction, stillborn and neonatal death. RESULTS: Three hundred and eighteen women were selected; forty-one women (12.9%) had PHS and 277 of them (87.1%) did not develop any of the alterations of the HELLP syndrome diagnosis. Preeclampsia was a more frequent type of hypertension in the PHS group than in the hypertension group. None of the women with isolated chronic hypertension developed PHS. The rate of cesarean delivery, eclampsia, and preterm delivery was significantly greater in the PHS group than in the hypertension group. CONCLUSION: We observed that aggressive procedures had been adopted for patients with PHS. These resulted in immediate interruption of pregnancy, with elevated cesarean rates and preterm delivery. Such decisions need to be reviewed, in order to reduce the cesarean rate and the incidence of preterm delivery. PMID- 12563426 TI - Staggered spondaic word test in epileptic patients. AB - CONTEXT: Auditory processing during childhood may be altered if there is any predisposing factor during the course of development. Neurological disorders are among the risk factors for auditory processing impairment. From this perspective, epileptic children present such a risk factor and could present auditory processing dysfunction. OBJECTIVE: To evaluate central auditory processing in epileptic patients using the Staggered Spondaic Word Test (SSW) in order to verify whether these patients presented auditory disorders and whether the type of crisis partial or generalized played a role in the occurrence and type of disorder. SETTING: Tertiary care hospital. SAMPLE: Thirty-eight children and adolescents, ranging from 7 to 16 years old, with a diagnosis of epilepsy divided into two groups: 23 patients with partial crisis and 15 patients with generalized crisis. MAIN MEASUREMENTS: Performance in the Staggered Spondaic Word Test versus epileptic crisis type (partial or generalized). RESULTS: The majority of epileptic patients showed central auditory processing disorders. There were no differences in relation to crisis type. Both groups showed similar performance, although the results observed for these patients differ from what is obtained with normal populations. With regard to response bias, there were also no differences in performance between subjects with partial or generalized seizures. All possible disorders were found in both groups, without the prevalence of one specific disorder over the other. CONCLUSIONS: This study revealed a high prevalence of disorders among epileptic patients in relation to processing partially overlapped verbal sounds in a dichotic paradigm. PMID- 12563427 TI - Hand-assisted bilateral nephrectomy in a patient with adult polycystic kidney disease. AB - CONTEXT: Dominantly autosomal polycystic disease is characterized by multiple bilateral and non-functional cysts, which lead to progressive kidney failure. OBJECTIVE: Our objective was to report on a case of hand-assisted bilateral nephrectomy in a 28-year-old female patient with adult polycystic disease and recurring pyelonephritis in a kidney transplant program. CASE REPORT: A hand assisted bilateral nephrectomy was performed through a supra-umbilical median incision of approximately 6 cm, and with 3 ports of 10 mm. The length of the surgery was 3 hours and 15 minutes. The kidneys were removed after the aspiration of some cysts through the supra-umbilical incision. Pain control was achieved via the use of analgesics. The blood loss during surgery was 160 ml. During the postoperative period, the patient developed right-side pneumothorax, which was drained with no further occurrence. This drain was kept in place for 48 hours. The length of hospitalization was 4 days. PMID- 12563428 TI - Agenesis of the gallbladder and cystic duct. AB - Agenesis of the gallbladder and cystic duct is a rare anomaly that is usually asymptomatic. The patient may present symptoms characteristic of cholelithiasis. Its surgical confirmation requires careful dissection of the common bile duct and intraoperative cholangiography or ultrasonography to be performed, to exclude the possibility of an ectopic gallbladder. The authors describe two cases of this unusual affection and comment on its clinical, pathophysiological and diagnostic aspects. PMID- 12563429 TI - Anterior cruciate ligament ganglion: case report. AB - CONTEXT: A ganglion is a cystic formation close to joints or tendinous sheaths, frequently found in the wrist, foot or knee. Intra-articular ganglia of the knee are rare, and most of them are located in the anterior cruciate ligament. The clinical picture for these ganglia comprises pain and movement restrictions in the knee, causing significant impairment to the patient. Symptoms are non specific, and anterior cruciate ligament ganglia are usually diagnosed through magnetic resonance imaging or arthroscopy. Not all ganglia diagnosed through magnetic resonance imaging need to undergo surgical treatment: only those that cause clinical signs and symptoms do. Surgical results are considered good or excellent in the vast majority of cases. CASE REPORT: A 29-year-old male presented with pain in the left knee during a marathon race. Physical examination revealed limitation in the maximum range of knee extension and pain in the posterior aspect of the left knee. Radiographs of the left knee were normal, but magnetic resonance imaging revealed a multi-lobed cystic structure adjacent to the anterior cruciate ligament, which resembled a ganglion cyst. The mass was removed through arthroscopy, and pathological examination revealed a synovial cyst. Patient recovery was excellent, and he resumed his usual training routine five months later. PMID- 12563450 TI - [Fibromyalgia: evidence of a neurophysiologic substrate]. PMID- 12563451 TI - [Authorship of scientific work]. PMID- 12563452 TI - [Predictive factors for the postoperative treatment of peptic ulcer hemorrhage]. PMID- 12563454 TI - [Hydrops fetalis in infant newborn with congenital cytomegaly ]. PMID- 12563456 TI - [Predictors of operative treatment in patients sustaining bleeding peptic ulcers]. AB - OBJECTIVE: To analyze the predictors of operative treatment in patients sustaining bleeding peptic ulcers. METHODS: Retrospective analysis of the patients admitted owing to bleeding peptic ulcers from 1998 to 2001. Vital signs upon admission, associated diseases, endoscopy data, evolution and definitive care were evaluated. Patients who underwent surgery were compared with the others in order to identify the predictors of the operative treatment. Student's t and chi squared test were employed for statistical analysis, considering p< 0,05 as significant. RESULTS: Two hundred patients were included in the study group. The age ranged between 17 and 97 years (mean 52 + 18), and 153 patients (76.5%) were male. Shock upon admission was present in 27 (13.5%). The endoscopic exam performed at admission showed that 101 patients sustained ulcers in the duodenum and 99 in the stomach, which were classified as Forrest Ia in 5 (2.5%), Ib in 20 (10%), IIa in 48 (24%) and IIb in 48 (24%). Twenty five (12.5%) patients needed operation, and 23 (92%) were submitted to gastrectomy with Billroth's II reconstruction. The operative treatment were more frequently performed in patients sustaining chronic alcoholic intake (p=0.002), cigarette smoking (p=0.02), diabetes mellitus (p=0.01), chronic peptic disease (p=0.05), shock upon admission (p<0.001) and spurting activity at the endoscopy (p<0.001). Forrest IIb lesions which did not receive any form of endoscopic treatment at admission were more likely to rebleed (p=0.012). CONCLUSIONS: The predictors of operative treatment were chronic alcoholic intake, smoke addiction, diabetes mellitus, chronic peptic disease, shock upon admission and Forrest Ia ulcers. PMID- 12563457 TI - [Neonatal and pediatric intensive care in Rio de Janeiro: distribution of beds and analysis of equity]. AB - OBJECTIVES: To identify the pediatric ICUs at Rio de Janeiro, number of beds, geographical distribution, public or private nature, type of hospital and assistance, studying population demand and to propose measures for improving equity. METHODS: All ICUs of the State were visited from July 97 to June 98, identifying number of beds and average length of stay. With this information along with demographic data from IBGE, the necessity of beds were estimated, comparing availability and demand by region, and proposing improving equity strategies. RESULTS: 80 ICUs were identified (6 excluded), totaling 1080 beds; 60% intensive and 40% semi-intensive; 57% public and 43% private; 52% in exclusive neonatal ICUs; 14% in pediatric and 34% in mixed (65% neonatal beds), totaling 791 neonatal beds (73%). The majority of ICUs (75%) were part of general hospitals, 20% were in obstetric or obstetric-pediatric hospitals, and only 5% were part of university centers; the majority were in metropolitan area (89%), with 93% of beds for 74% of state children population, of whom, the majority were in Rio de Janeiro city (76%), with 73% of beds for 37% of population, contrasting with the inner of the State, with only 8 units (11%) and 79 beds (7%) for 26% of children. CONCLUSIONS: There is no equity in the distribution and accessibility to the available beds, with lack in public and excess in private sectors, a great concentration in the metropolitan area and only 5% of ICUs at university hospitals, recommending a policy of redistribution and allocation of new beds in more needy areas, associated with the creation of an admission center along with an efficient reference and transportation system. PMID- 12563458 TI - [Effect of acetylsalicylic acid on the reproductive performance and on offspring from wistar rats]. AB - The aim of this paper was to perform a randomized, controlled and blinded study to investigate if a therapeutic dose of acetylsalicylic acid (ASA), taken by pregnant women, may also cause embryotoxic or congenital abnormalities on experimental animal. METHODS: Females were confirmed to have mated by observations of sperm in a vaginal smear. The day on which spermatozoa were found in the vaginal smear was considered as day 1 of gestation (GD1). After randomization, mated females were assigned to experimental groups and individually caged, were given 50 mg/kg/day of acetylsalicylic acid, by needle gavage once daily, during two different periods of pregnancy. One group of dams (n=11) received aspirin from day 1 to 4 of pregnancy (before embryonic implantation) for evaluation of the blastocysts, and another group received aspirin from day 6 to 15 of pregnancy (organogenic period) for fetal evaluation. Control groups (n=12) received distilled water in same volume and during same periods as their respective experimental groups. RESULTS AND CONCLUSION: The treatment of the dams with ASA, according to minimal therapeutic dose used for humans, did not cause embryotoxic or major malformations on experimental animal but was responsible for rate increased of fetuses presenting ureteric dilatation. After analysis of the data, it appears that, although direct conclusive evidence of adverse effects in humans is lacking, a potential hazard dose exists and thus the indiscriminate use of acetylsalicylic acid (aspirin) is contraindicated. PMID- 12563459 TI - [Relationship between the outcome and the frequency of micronuclei in cells of patients with oral and oropharyngeal carcinoma]. AB - BACKGROUND: To verify the correlation between the micronucleus frequency and the presence of local recurrences or second primary lesion in patients with carcinoma of the oral cavity. METHODS: We studied the frequency of micronucleus of the oral mucosa in 27 untreated patients with carcinoma of the oral cavity and oropharynx. The patients were monthly followed after initial treatment, in an attempt to identify local recurrences or second primary lesions. RESULTS: Of the 24 patients evaluated during the whole time, 5 cases (20,8%) were alive and free of disease, and 19 cases died, 18 (75%) owing to cancer and 1 to perfurative peptic ulcer. In relation to micronucleus frequency, no difference was observed among the patients with local recurrence of the disease (N=14) and the patients who died of other causes or were alive and free of disease (N=10) (p = 0.83). There was higher micronucleus frequency in the stages T3 and T4 (p = 0.01). CONCLUSION: In the present study was not possible to find a clinical correlation between the frequency of micronucleus of the oral mucosal and the risk of development of local recurrence or second primary tumor in patients with upper aerodigestive tract carcinoma. PMID- 12563460 TI - [Oximetry of stomach: comparative study between proximal gastric vagotomy and anterior lesser curve seromyotomy with posterior truncal vagotomy in chronic duodenal ulcer]. AB - Delayed gastric emptying of solid food is greater after proximal gastric vagotomy (PGV) than after anterior lesser curve seromyotomy with posterior truncal vagotomy (ASPTV) and may be due to the hypoxia in the gastric wall and mainly in the gastric pacemaker. PURPOSE: To verify if operative devascularization of the lesser curve and the gastric fundus could affect the entire stomach and particularly the pacemaker area. METHODS: Measures of intraoperative oxyhemoglobin saturation (SpO2) were taken by pulse oximetry on the anterior gastric wall in 20 patients with chronic duodenal ulcer, randomly allocated in two groups of 10 individuals for surgical treatment, by PGV or ASPTV. RESULTS: Measurements, before partial fundoplication, showed that the area of the proximal lesser curve and the gastric fundus had a significant decrease in the SpO2 when compared to the gastric corpus (p < 0,05); the SpO2 was not reduced significantly by the procedures on the pacemaker area; the ligature of short gastric vessels, in association with PGV or ASPTV, reduced significantly the SpO2 (p < 0,05); and, PGV resulted in a SpO2 significantly lower than in ASPTV (p < 0,05). CONCLUSION: When PGV is associated with the ligature of the short gastric vessels, it causes acute ischemic changes in the proximal lesser gastric curve and in the fundus more intensively than ASPTV. The gastric pacemaker area does not present hypoxia immediately after operative procedures. PMID- 12563461 TI - [Latent TGFbeta1 activation in the lung irradiated in vivo]. AB - PURPOSE: assess the latent and active TGFb1 localization in the lung, whether or not radiation induces latent TGFbeta1 activation, and the distribution of collagen fibers in the irradiated lung. METHODS: Thirty two C57BL mice were randomly assigned in two groups: GI (non irradiated animals) and GII (irradiated animals). The mice from GII received a single whole - body radiation dose of 7Gy, using a 60Co source at a dose rate of 0.97 Gy/min. They were sacrificed by cervical dislocation at 1, 14, 30 and 90 days after radiation. RESULTS: The irradiated lungs showed: 1) vascular congestion and thickness of the alveolar septa 30 days and more intense 90 days after irradiation; 2) significant increase of collagen deposition in all time periods after irradiation; 3) weak latent TGFbeta1 activation 1 day and strong activation 14 days after irradiation in the bronchi and alveoli. Our results suggest that some bronchial and alveolar cells may have a role in the complex process of radiation-induced lung fibrosis acting as cellular sources of active TGFbeta. PMID- 12563462 TI - [Treatment of autonomous and cystic thyroid nodules with intranodular ethanol injection]. AB - Intranodular ethanol injection has been used for the past 10 years as an efficient modality for treating patients with thyroid nodules. Several studies have reported the success of this therapy in autonomous and cystic nodules and, more recently, in cold benign nodules. PURPOSE: To evaluate the efficacy of this therapeutic modality on the treatment of autonomous and cystic thyroid nodules. METHODS: 42 patients (26 with cystic and 16 with autonomous nodules) were treated with ultrasound guided intranodular 99% ethanol injection and followed for 6 months. RESULTS: No major complications were observed during or after treatment, however, most of the patients reported slight to moderate pain and/or discomfort after the injection. Most of the nodules showed reduction after the treatment. Autonomous nodules had a mean reduction of 50.3% and cystic nodules of 69.3%. No significant differences in pretreatment serum total T3, total T4 or TSH were observed among the patients in the cystic group. Patients in the autonomous group with hyperfunctioning nodules showed a decrease in serum total T3, total T4 and an increase in serum TSH levels, hence, proving the effectiveness of this therapy. CONCLUSIONS: Intranodular ethanol injection is a safe and efficient treatment for autonomous and cystic nodules of the thyroid. PMID- 12563463 TI - [Squamous cell carcinoma of the mouth in the elderly in Sao Paulo]. AB - OBJECTIVE: To assess the influence of sex and primary tumor location on the outcome of mouth cancer in elderly patients. METHODS: A retrospective study of 1,440 patients with squamous cell carcinoma of mouth in the Head and Neck Department of Heliopolis Hospital, Sao Paulo during 1978-1997 period. There were 562 elderly patients (aged 60 or more years, according to WHO criteria for developing countries) and 878 patients younger than 60 years old. The variables sex, location of the tumor and life time after detection were compared between the two groups of patients. The chi squared test (x2 ) was used for statistical analysis with a 0.05 significance level (confidence interval of 95%). RESULTS: The frequency of the mouth cancer in elderly patients remained stable in the studied period (39,5% in 1978-87 versus 38.2% in 1988-1997). The male/female ratio in squamous cell carcinoma of the oral cavity changed from 3:1 before 60 years old to 8:1 after this age. There was a predominance in buccal mucosa (56%) and palate (47%) in aged people when compared with the tumors of younger patients preferentially in floor/tongue (67%) and tongue (62%). No differences were found in the percentage of deceased patients before the beginning of therapeutic procedures (11.6% versus 10.5% in younger) or in the percentages of survivors, in different periods (6 months the five years). CONCLUSIONS: A higher frequency of women with mouth cancer was observed in the elderly patients. The upper floor of the mouth site was more frequent in the patients aged 60 years or more while the inferior location (tongue and floor) were more frequent in patients under 60. The observed differences in relation to patient's sex and tumor location did not influence outcome in mouth cancer patients. PMID- 12563464 TI - [Analysis of coagulation parameters in patients undergoing controlled ovarian hyperstimulation for in vitro fertilization]. AB - BACKGROUND: Hyperestrogenic conditions have been related to alterations in the coagulation parameters. The objective of this study was to identify changes in coagulation parameters in women undergoing in vitro fertilization and embryo transfer (IVF-ET). METHODS: A group of 15 patients was studied prospectively, immediately before and during the course of an IVF-ET cycle. Blood was drawn in the preovulatory period of the cycle preceeding IVF-ET for complete blood count, fibrinogen, prothrombin time (TP), thromboplastin partial actived time (ATTP), coagulation time and estradiol. A another blood sample for the same tests was collected during controlled ovarian hyperstimulation, on the hCG day. Both samples were compared by a paired t-test. RESULTS: There was an increase in fibrinogen (340.9 pg/mL to 383.1 pg/mL, p = 0.01). On the other hand, hematocrit levels decreased (40.5% to 38.4 %, p = 0.0001). The other coagulation tests showed no difference. Estradiol levels increased (167 pg/mL to 1435 pg/ml, p=0.0001), as expected during such treatment. CONCLUSIONS: The data suggest a tendency towards an increase in coagulability, as expected in hyperestrogenic situations. However, even considering the statistical significance of the results, they might have limited clinical impact, since they were within the normal range. This study suggests that, despite the marked increase in estradiol and changes in coagulation parameters, IVF-ET is a safe procedure in terms of thromboembolic risks. PMID- 12563465 TI - [Early feeding after intestinal anastomoses: risks or benefits?]. AB - BACKGROUND: Oral intake after intestinal anastomoses has traditionally been prescribed only after the resolution of ileus. The aim of this study was to evaluate the immediate results of early oral feeding in postoperative course of intestinal anastomoses. METHODS: 43 consecutive patients submitted to operations associated with intestinal anastomoses were randomized to two groups: early group (N=23) allowed to oral intake after 24 h of operation and conventional group (N=20) in which feeding was allowed only after the ileus resolution. Groups were statistically similar for age, gender, nutritional status, duration of operation, local and type of anastomoses, type of anesthesia and use of morphine. RESULTS: Three (13%) patients of early group and two (10%) of conventional presented vomiting requiring nasogastric decompression (P.0.05). Three (7.3%) patients, two (10%) in early feeding group and one (5%) in conventional group died (P>0.05). There was no difference between the groups for morbidity. Two anastomotic dehiscences occurred in each group. Passage of flatus was reported earlier (P=0.01) in early feeding group (35 +/- 13 h) when compared with conventional group (44 +/- 12 h). Hospital stay was similar between the two groups (conventional group = 12 [6-36] days versus early feeding group = 10 [5-29] days). CONCLUSIONS: The return of oral feeding on the first postoperative day in patients submitted to intestinal anastomoses is safe, not associated with the occurrence of anastomotic dehiscence and moreover, related to a shorter resolution of ileus. PMID- 12563466 TI - [Distribution of the lateral cutaneous nerve of the thigh in the area of intramuscular injection]. AB - The technique of intramuscular injection (IM) into the antero-lateral region of the thigh is widely used. Nevertheless, despite this area being indicated as the second best location for this practice, the technique is still observed to be very painful for both adult and child patients. OBJECTIVE: To study the localization, distribution and course of the lateral cutaneous nerve of the thigh, and its topographic relationship with the area recommended for the practice of intramuscular injection, relating these characteristics to the pain resulting from such procedures. METHOD: By means of exposing the antero-lateral region by classical dissection, the lateral cutaneous nerve of the thigh was identified and isolated in 20 fixed adult male cadavers, giving emphasis to the viewing of its nerve rami across the iliotibial tract. RESULTS: In 100% of the cases, the lateral cutaneous nerve emerged medially in relation to the upper anterior iliac spine. After this, it issued three wide-caliber rami in 70% of the specimens and only two in the remaining 30%. In the upper third and in the upper portion of the middle third of the thigh, a network of numerous small nerve rami was observed, enveloped in a variable quantity of adipose tissue. However, in the lower portion of the middle third of the thigh and in the lower third, no significant nerve rami were seen. CONCLUSION: Based on our data, we recommend whenever possible that the distal half of the region displayed by the classical technique be utilized as the location of choice for the practice of intramuscular injection into the antero-lateral region of the thigh. This is because this region is less innervated by the lateral cutaneous nerve of the thigh, which will cause less pain in this area during such procedures, thereby affording greater comfort to the patient. PMID- 12563467 TI - [Smallpox, vaccine prevention and the bioterrorism threat]. AB - With the international threat of bioterrorism, smallpox is again a matter of intense debate. The authors review this disease and raise considerations on the exploitation of the etiologic agent as a biological weapon. Also bring to discussion the available efforts to produce and develop vaccines against the disease. PMID- 12563468 TI - Prevalence of Chlamydia trachomatis in human immunodeficiency virus-infected women in Cuba. AB - To determine the prevalence rates and serovar distribution of Chlamydia trachomatis cervical infections in Cuban women, two different groups were selected. Group I consisted of 60 human immunodeficiency virus (HIV-1) seropositive women from different regions of Cuba and group II of 60 randomly selected women HIV seronegative and apparently healthy. C. trachomatis was detected in cervical scrapes by mean of nested polymerase chain reaction (PCR) specific for major out membrane protein. The overall prevalence rate of C. trachomatis in cervical scrapes determined by nested PCR was 10% in group I and the estimated prevalence was 6.6% for group II; 83.3% of HIV seropositive women with C. trachomatis infection reported history of pelvic inflammatory disease followed by cervicitis (50%). The control group C. trachomatis-infected women referred a history of cervicitis in 75% of cases. Other reports in the latter group included infertility and pelvic inflamatory disease in 50%. The present study is the first report of C. trachomatis prevalence in Cuba. It showed that there was not significantly difference in the prevalence rate of C. trachomatis between both groups. PMID- 12563469 TI - Epidemiological and immunological aspects of human visceral leishmaniasis on Margarita Island, Venezuela. AB - Sixty-five patients were diagnosed with visceral leishmaniasis (VL) on Margarita Island in the decade from 1990 to1999; 86.2% were <= 3 years old. All were leishmanin-negative at diagnosis. Evaluation of 23 cured patients in 1999 revealed that 22/23 had converted to leishmanin-positive; five had persisting antibodies to rK39 antigen, with no clinical evidence of disease. Leishmanin tests were positive in 20.2% of 1,643 healthy individuals from 417 households in endemic areas. Of the positive reactors, 39.8% were identified in 35 (8.4%) of the households, 15 of which had an antecedent case of VL, a serologically positive dog or both. Weak serological activity to rK39 antigen was detected in 3 of 488 human sera from the endemic areas. The presence of micro-foci of intense peri-urban transmission and the apparent absence of other Trypanosomatidae causing human disease offer a unique opportunity for the study of reservoirs, alternative vectors and evaluation of control measures on the Island. PMID- 12563470 TI - Phenotypic and genotypic characteristics of shiga toxin-producing Escherichia coli strains isolated from children in Sao Paulo, Brazil. AB - The biochemical and serological characteristics, virulence properties, and genetic relatedness of Shiga toxin-producing Escherichia coli (STEC) strains isolated in S o Paulo, from April 1989 through March 1990, were determined. This is also the first report on clinic findings of human STEC infections in Brazil. The only three STEC strains identified in that period were lysine decarboxylase negative, belonged to serotype O111ac: non-motile, were Stx1 producers, carried the eae and astA genes, and 2 of them also presented the EHEC-hly sequence. The children carrying STEC were all boys, with less than two years old, and had no previous history of hospitalization. None of them presented blood in stools. Vomiting, cough and coryza were the most common clinical manifestations observed. Although the STEC strains were isolated during summer months, and presented similar phenotypic and genotypic characteristics, carbohydrate fermentation patterns and PFGE analysis suggested that these diarrheal episodes were not caused by a single clone. PMID- 12563471 TI - Humoral immune responses induced by Kudoa sp. (Myxosporea: Multivalvulida) antigens in BALB/c mice. AB - The majority of Kudoa species infect the somatic muscle of fish establishing cysts. As there is no effective method to detect infected fish without destroying them these parasited fish reach the consumer. This work was developed to determine whether this parasite contains antigenic compounds capable of provoking an immune response in laboratory animals, in order to consider the possible immunopathological effects in man by the ingestion of Kudoa infected fish. BALB/c mice were injected by the subcutaneous route with the following extracts suspended in aluminium hydroxide: group 1 (black Kudoa sp. pseudocyst extract), group 2 (white Kudoa sp. pseudocyst extract), and group 3 (non-infected hake meat extract). Specific antibody levels were measured by ELISA against homologous and heterologous antigens. The highest responses were obtained from the black Kudoa sp. pseudocyst extract (group 1). The low optic density levels detected in group 3 proved that the results obtained in groups 1 and 2 were a consequence of the parasitic extract injection. The IgG1 was the predominant subclass. IgE detected in groups 1 and 2 showed the possible allergenic nature of some of the components of the parasitic extract. High IgA levels and medium IgG2a and IgG3 levels were obtained in groups 1 and 2. Low IgG2b responses were shown. No cross-reactions between Kudoa sp. pseudocyst extracts and the non-infected hake meat extract were observed. PMID- 12563472 TI - Kinetics of T cell-activation molecules in response to Mycobacterium tuberculosis antigens. AB - The phenotypic features acquired subsequent to antigen-specific stimulation in vitro were evaluated by means of the kinetic expressions of CD69 and CD25 activation molecules on T lymphocytes and assayed by flow cytometry in response to PPD, Ag85B, and ferritin in PPD-positive healthy control individuals. In response to PHA, CD69 staining on both CD4+ and CD8+ T cells became initially marked after 4 h, peaked at 24 h, and quickly decreased after 120 h. For CD25, a latter expression was detected around 8 h, having increased after 96 h. As expected, the response rate to the mycobacterial antigens was much lower than that to the mitogen. Positive staining was high after 96 h for CD25 and after 24 h for CD69. CD69 expression was significantly enhanced (p < 0.05) on CD8+ as compared to CD4+ T cells. High levels were also found between 96-120 h. Regarding Ag85B, CD25+ cells were mostly CD4+ instead of CD8+ T cells. Moreover, in response to ferritin, a lower CD25 expression was noted. The present data will allow further characterization of the immune response to new mycobacterial specific antigens and their evaluation for possible inclusion in developing new diagnostic techniques for tuberculosis as well in a new vaccine to prevent the disease. PMID- 12563473 TI - Biological characterization of a beta-galactosidase expressing clone of Trypanosoma cruzi CL strain. AB - Clone CL B5 of Trypanosoma cruzi is a beta-galactosidase expressing organism that was genetically transfected to be used for in vitro pharmacological screening. Biological parameters were determined, evaluating growth kinetics of epimastigotes, metacyclogenesis, infectivity to mammalian cell lines, parasitemia kinetics in mice and sensibility to nifurtimox and benznidazole. Differences in relation to other strains and CL parental strain were found, the most important being the incapability to produce death to mice in spite of the high inoculum used. However, it possesses the required features to be used for in vitro drug screening. Data obtained demonstrate that heterogeneity of T. cruzi appears even among clones of the same strain, and that these differences found do not prevent the use of clone CL B5 for the purpose that was engineered. PMID- 12563474 TI - Isolation of fungi in Musca domestica Linnaeus, 1758 (Diptera: Muscidae) captured at two natural breeding grounds in the municipality of Serop dica, Rio de Janeiro, Brazil. AB - The objective of this study was to isolate and identify fungal species found in natural association with adults of Musca domestica. The adult insects were collected from two natural breeding grounds: hog pens and an urban sanitary landfill. The isolated fungi were identified as: Aspergillus flavus (23.8%), A. niger var. niger (14.4%), Penicillium corylophilum (21.4%), P. fellutanum (11.9%), Cladosporium cladosporoides (4.7%), Fusarium sp. (4.7%), Alternaria alternata (11.9%), Curvularia brachyspora (2.4%), Mycelia sterilia (2.4%) and the Mucorales order (2.4%). PMID- 12563475 TI - Effect of sequential heat and cold shocks on nuclear phenotypes of the blood sucking insect, Panstrongylus megistus (Burmeister) (Hemiptera, Reduviidae). AB - Thermal shocks induce changes in the nuclear phenotypes that correspond to survival (heterochromatin decondensation, nuclear fusion) or death (apoptosis, necrosis) responses in the Malpighian tubules of Panstrongylus megistus. Since thermal tolerance increased survival and molting rate in this species following sequential shocks, we investigated whether changes in nuclear phenotypes accompanied the insect survival response to sequential thermal shocks. Fifth instar nymphs were subjected to a single heat (35 or 40 degrees C, 1 h) or cold (5 or 0 degrees C, 1 h) shock and then subjected to a second shock for 12 h at 40 or 0 degrees C, respectively, after 8, 18, 24 and 72 h at 28 degrees C (control temperature). As with specimen survival, sequential heat and cold shocks induced changes in frequency of the mentioned nuclear phenotypes although their patterns differed. The heat shock tolerance involved decrease in apoptosis simultaneous to increase in cell survival responses. Sequential cold shocks did not involve cell/nuclear fusion and even elicited increase in necrosis with advancing time after shocks. The temperatures of 40 and 0 degrees C were more effective than the temperatures of 35 and 5 degrees C in eliciting the heat and cold shock tolerances, respectively, as shown by cytological analysis of the nuclear phenotypes. It is concluded that different sequential thermal shocks can trigger different mechanisms of cellular protection against stress in P. megistus, favoring the insect to adapt to various ecotopes. PMID- 12563476 TI - Temperature influence on embryonic development of Anopheles albitarsis and Anopheles aquasalis. AB - Temperature influence on the embryonic development of Anopheles aquasalis and An. albitarsis was investigated. At 26 degrees C, 75% and 60% of respectively An. aquasalis and An. albitarsis eggs hatched, with one peak of eclosion, between the 2nd and 3rd day after oviposition. At 20 +/- 2 degrees C, around 66-70% of An. aquasalis eggs hatched, with one eclosion peak, on the 5th day. On the other hand, An. albitarsis eclosion at 21+/- 2 degrees C decreased to 10-22%, with two eclosion peaks, on the 4th-5th day and on the 9th-12th day. These data indicate a stronger temperature influence over An.albitarsis than over An. aquasalis embryos. PMID- 12563477 TI - Relationship between antennal sensilla pattern and habitat in six species of Triatominae. AB - In order to determine if habitat similarity is correlated with a similarity of sensilla pattern, we analyzed six species of Triatominae present in two biogeographic regions of Brazil: the "caatinga" and the "cerrado". In broad terms Triatoma infestans (cerrado) and T. brasiliensis (caatinga) are found in human domiciles, T. sordida (cerrado) and T. pseudomaculata (caatinga) colonize peridomestic habitats, and Rhodnius neglectus (cerrado) and R. nasutus (caatinga) inhabit palm tree crowns. The number and distribution of four sensilla types (bristles, thin and thick walled trichoidea, and basiconica) were compared in these species. Sexual dimorphism of sensilla patterns was noted in T. sordida, T. brasiliensis and T. pseudomaculata. A principal component analysis showed three main groups: (i) species that live in the palms, (ii) domiciliated species and (iii) those living in the peridomestic habitat. T. infestans almost exclusively domestic, was placed at the centre of the canonical map and some individuals of other species overlapped there. These results support the idea that the patterns of antennal sensilla are sensitive indicators of adaptive process in Triatominae. We propose that those species that inhabit less stable habitats possess more types of sensilla on the pedicel, and higher number of antennal sensilla. PMID- 12563478 TI - Triatoma rubrovaria (Blanchard, 1843) (Hemiptera, Reduviidae, Triatominae) II: trophic resources and ecological observations of five populations collected in the State of Rio Grande do Sul, Brazil. AB - Triatoma rubrovaria has become the most frequently captured triatomine species after the control of T. infestans in the State of Rio Grande do Sul (RS), Brazil. Isoenzymatic and chromatic studies indicate the existence of, at least, two distinct phenotypic patterns of T. rubrovaria in RS. The geographic variation noted through molecular tools may also result in distinct profiles of vectorial potentiality. In order to enhance our understanding of the bionomic knowledge of T. rubrovaria separate batches of the species were collected from different municipalities of RS distant from 72 to 332 km: Santana do Livramento (natural ecotope), Santana do Livramento (artificial ecotope), Santiago (natural ecotope), Cangu u (peridomicile) and Encruzilhada do Sul (natural ecotope). A total of 285 specimens were collected, 85 specimens kept sufficient fecal material in their guts for the precipitin analysis. The results indicated the food eclecticism for this species and the anti-rodent serum showed the highest positivity in most localities. From the total of analyzed samples, only 1.3% of unique positivity for human blood was registered, all of them for Santiago population. This reactivity to human blood may be associated to pastures activities in the field. PMID- 12563479 TI - Ticks (Acari: Ixodidae) on wild animals from the Porto-Primavera Hydroelectric power station area, Brazil. AB - From June 2000 to June 2001, a total of 741 ticks were collected from 51 free living wild animals captured at the Porto-Primavera Hydroelectric power station area, located alongside an approximately 180 km course of the Paran river, between the states of S o Paulo and Mato Grosso do Sul, comprising 9 species of 3 genera: Ambly-omma (7 species), Boophilus (1) and Anocentor (1). A total of 421 immature Amblyomma ticks were reared in laboratory until the adult stage, allowing identification of the species. A. cajennense was the most frequent tick species (mostly immature stages) collected on 9 host species: Myrmecophaga tridactyla, Tamandua tetradactyla,Cerdocyon thous, Puma concolor,Tayassu tajacu, Mazama gouazoubira,Hydrochaeris hydrochaeris,Alouatta caraya, Cebus apella. Other tick species were less common, generally restricted to certain host taxa. PMID- 12563480 TI - Larval dispersal and predation in experimental populations of Chrysomya albiceps and Cochliomyia macellaria (Diptera: Calliphoridae). AB - In this study we investigated the larval dispersal associated with larval predation in experimental populations of Chrysomya albiceps and Cochliomyia macellaria. Frequency distribution of sampling units (G test) in the substrate was used to evaluate variation in larval dispersal. An experimental acrylic channel (1 x 0.1 x 0.2 m) covered with wood shavings was used to observe larval dispersal prior to pupation. The acrylic channel was graduated at 0.05 m intervals, each representing a sampling unit; hence, 20 sampling units were set up. A Petri dish containing third instar larvae of single and double species was deposited at one edge of the acrylic channel allowing larvae to disperse. The number of buried pupae (0, 1, 2, n) present in each sampling unit was recorded. For double species, the number of recovered larvae of C. albiceps was similar to the number initially released on the dish Petri. On the other hand, the number of recovered larvae of C. macellaria was significantly smaller than the initially released number. The results show that C. albiceps attacks C. macellaria larvae during the larval dispersal process. The larval distribution of C. albiceps did not differ significantly from C. macellaria in double species, but it differed significantly in single species. The larval aggregation level of C. macellaria decreased when C. albiceps was present and the larval aggregation level of C. albiceps increased when C. macellaria was present. The implications of such findings for the population dynamics of these species are discussed. PMID- 12563481 TI - Brazilian distribution of Amblyomma varium Koch, 1844 (Acari: Ixodidae), a common parasite of sloths (Mammalia: Xenarthra). AB - Amblyomma varium, commonly known in Brazil as the "carrapato-gigante-da-pregui a" (sloth's giant tick) is found from southern Central America to Argentina. The present study adds information on the geographical distribution of A. varium, as well as on their hosts, based on material deposited in the main Brazilian collections and on the available literature. Eighty-two vials, containing 191 adult specimens, deposited in five Acari collections between 1930 and 2001, were examined. These vials included data on the host and collection localities. The biology of A. varium is unknown. However it is known that, during the adult stage, the tick presents a high host specificity and is found almost exclusively on the sloths Bradypus tridactylus, B. variegatus, B.torquatus (Bradypodidae), Choloepus hoffmanni and C. didactylus (Megalonychidae). Based on the material examined, the states of Rond nia, Amazonas, Bahia and Alagoas are newly assigned to geographic distribution of A. varium in Brazil. PMID- 12563482 TI - Drug resistance and genotypes of strains of Mycobacterium tuberculosis isolated from human immunodeficiency virus-infected and non-infected tuberculosis patients in Bauru, Sao Paulo, Brazil. AB - Little is known about transmission and drug resistance of tuberculosis (TB) in Bauru, State of S o Paulo. The objective of this study was to evaluate risk factors for transmission of Mycobacterium tuberculosis strains in this area. Strains were collected from patients attended at ambulatory services in the region and susceptibility towards the main first line antibiotics was determined and fingerprinting performed. A total of 57 strains were submitted to susceptibility testing: 23 (42.6%) were resistant to at least one drug while 3 (13%) were resistant against both rifampicin and isoniazide. Resistant strains had been isolated from patients that had not (n = 13) or had (n = 9) previously been submitted to anti-TB treatment, demonstrating a preoccupying high level of primary resistance in the context of the study. All strains were submitted to IS6110 restriction fragment length polymorphism (IS6110-RFLP) and double repetitive element PCR (DRE-PCR). Using IS6110-RFLP, 26.3% of the strains were clustered and one cluster of 3 patients included 2 HIV-infected individuals that had been hospitalized together during 16 days; clustering of strains of patients from the hospital was however not higher than that of patients attended at health posts. According to DRE-PCR, 55.3% belonged to a cluster, confirming the larger discriminatory power of IS6110-RFLP when compared to DRE-PCR, that should therefore be used as a screening procedure only. No clinical, epidemiological or microbiological characteristics were associated with clustering so risk factors for transmission of TB could not be defined in the present study. PMID- 12563483 TI - Detection of hilA gene sequences in serovars of Salmonella enterica subspecies enterica. AB - hilA gene promoter, component of the Salmonella Pathogenicity Island 1, has been found in Salmonella serovar Typhimurium, being important for the regulation of type III secretion apparatus genes. We detected hilA gene sequences in Salmonella serovars Typhi, Enteritidis, Choleraesuis, Paratyphi A and B, and Pullorum, by polymerase chain reaction (PCR) and hybridization techniques. The primers to carry out PCR were designed according to hilA sequence. A low stringency hybridization with the probe pVV441 (hilA open-reading-frame plasmid) was carried out. To find hilA gene sequences in other Salmonella sp. suggest that these serovars could have similar sequences of this kind of virulence genes. PMID- 12563484 TI - Mtp-40 and alpha antigen gene fragment amplification for the detection of Mycobacterium tuberculosis in Colombian clinical specimens. AB - In this study, the use of Mtp-40 and alpha antigen polymerase chain reaction (PCR) amplification fragments for the precise tuberculosis (TB) diagnosis was evaluated. One hundred and ninety two different samples were obtained from 113 patients with suspected TB. Mtp-40 and alpha antigen protein genes were amplified by the PCR technique and compared to both the "gold standard" (culture) test, as well as the clinical parameters (including a clinical record and X-ray film exam in 113 patients). Thirty-eight of the 113 patients had a presumptive clinical diagnosis of TB; 74% being detected by PCR technique, 58% by culture and 44% by direct microscopic visualization. Weconclude that it is possible to use PCR as a suitable technique for the detection of any mycobacteria by means of the alpha antigen product, or the specific infection of Mycobacterium tuberculosis by means of the mtp-40 gene. This might be a good supporting tool in difficult clinical TB diagnosis and pauci-bacillary cases. PMID- 12563485 TI - Detection of Giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies. AB - The present study developed and standardized an enzime-linked immunosorbent assay (ELISA) to detect Giardia antigen in feces using rabbit polyclonal antibodies. Giardia cysts were purified from human fecal samples by sucrose and percoll gradients. Gerbils (Meriones unguiculatus) were infected to obtain trophozoites. Rabbits were inoculated with either cyst or trophozoite antigens of 14 Colombian Giardia isolates to develop antibodies against the respective stages. The IgG anti-Giardia were purified by sequential caprylic acid and ammonium sulfate precipitation. A portion of these polyclonal antibodies was linked to alkaline phosphatase (conjugate). One hundred and ninety six samples of human feces, from different patients, were tested by parasitologic diagnosis: 69 were positive for Giardia cysts, 56 had no Giardia parasites, and 71 revealed parasites other than Giardia. The optimal concentration of polyclonal antibodies for antigen capture was 40 g/ml and the optimal conjugate dilution was 1:100. The absorbance cut-off value was 0.24. The parameters of the ELISA test for Giardia antigen detection were: sensitivity, 100% (95% CI: 93.4-100%); specificity, 95% (95% CI: 88.6 97.6%); positive predictive value, 91% (95% CI: 81.4-95.9%); and negative predictive value, 100% (95% CI: 96.1-100%). This ELISA will improve the diagnosis of Giardia infections in Colombia and will be useful in following patients after treatment. PMID- 12563486 TI - Morphologic and morphometric analysis of Hepatozoon spp. (Apicomplexa, Hepatozoidae) of snakes. AB - Hepatozoon species are the most abundant hemoparasites of snakes. Its identification has been based mainly on the morphologic characterization of the gamonts in the peripheral blood of the vertebrate host and also of the cysts found in the internal organs of the vertebrate and invertebrate hosts. Using a computerized image analysis system, we studied five species of Hepatozoon from recently captured snakes in Botucatu, State of S o Paulo, Brazil, to evaluate the importance of the morphology and morphometry of the gamonts for the characterization of Hepatozoon species and to analyze the morphologic changes induced in the erythrocytes by the parasite. The studied species were H. terzii of Boa constrictor amarali, Hepatozoon sp. of Crotalus durissusterrificus, H. philodryasi of Philodryas patagoniensis, and H. migonei and H. cyclagrasi of Hydrodynastes gigas. We observed three different groups, one of them including the species H. terzii, H. philodryasi and Hepatozoon sp. of C. durissus terrificus; and the other two consisting of H. migonei and H. cyclagrasi. Degree of alterations in the erythrocytes was variable and it may be useful for characterization of Hepatozoon species. PMID- 12563487 TI - Systematic studies on Anopheles galvaoi Causey, Deane & Deane from the subgenus Nysssorhynchus blanchard (Diptera: Culicidae). AB - Anopheles galvaoi, a member of the subgenus Nyssorhynchus, is redescribed based on morphological characters of the adults male and female, fourth-instar larva and pupa. Female, male genitalia, larval and pupal stages are illustrated. Data about medical importance, bionomics, and distribution are given based on literature records. Adult female of An. galvaoi can be easily misidentified as An. benarrochi Gabald n and An. aquasalis Curry. A few characters are indicated for identifying female and immatures of An. galvaoi. Phylogenetic relationships among An. galvaoi and six other species of the Oswaldoi Subgroup are estimated using COII mtDNA and ITS2 rDNA gene sequences. Lectotype of An. galvaoi, an adult female from Rio Branco, State of Acre, is invalidated. PMID- 12563488 TI - Morphometric discrimination of females of five species of Anopheles of the subgenus Nyssorhynchus from Southern and Northwest Colombia. AB - The most important vectors of human Plasmodium in the neotropics belong to the subgenus Nyssorhynchus. These species are generally sympatric in terms of their geographical distributions. Some are difficult to identify based solely on examination of adult females using the available morphological keys, in these cases examination of immature stages and male genitalia is required to make correct determinations. However, in epidemiological studies it is necessary to identify the species of adult females which are found near humans, i.e. in studies of malaria transmission or evaluation of control measures. The purpose of the present study was to evaluate the discrimination of adult females of different species of Nyssorhynchus isolated mainly from Southern Colombia (department of Putumayo), using morphometric analysis. Adult females were obtained after rearing larvae collected in natural breeding places and from the progeny of females collected on humans. The morphological characteristics of the immature stages allowed the identification of four species of the subgroup Oswaldoi from Southern Colombia: Anopheles rangeli Gabaldon, Cova Garcia & Lopez, An. oswaldoi (Peryassu), An. benarrochi Gabaldon, Cova Garcia & Lopez and An. triannulatus (Neiva & Pinto). The species An. nuneztovari (Gabaldon) from the Northwest of Colombia was included for comparison. Morphometric analysis allowed differentiation of the females of all species to a confidence level approaching 90% using principal components analysis of 10 wing and leg variables, followed by canonical variate analysis of the first four principal components. We conclude that morphometrics may represent a useful taxonomic tool for this group and that its use should be further studied. PMID- 12563489 TI - A new species of Anoplodiscus (Monogenea: Anoplodiscidae) parasitic on Pagrus pagrus (Osteichthyes: Sparidae) from the coastal zone of the State of Rio de Janeiro, Brazil. AB - A new species of Anoplodiscus (Monogenea, Anoplodiscidae), parasitic on gills of the red porgy, Pagrus pagrus, from the coastal zone of the State of Rio de Janeiro, Brazil, is described and illustrated. The new species can be differentiated from the other species of this genus by the shape of the accessory piece of the copulatory complex, and the length of the vagina. This is the first record of a species of Anoplodiscus in the Neotropical region. PMID- 12563490 TI - Acute inflammatory response in the stomach of BALB/c mice challenged with coccoidal Helicobacter pylori. AB - An experimental murine model was used to verify the viability and pathogenicity of coccoid Helicobacter pylori. For this purpose, 27 BALB/c mice were inoculated intragastrically with 1 ml broth culture (10(8)organisms/ml) of a coccoid H. pylori clinical isolate. The animals were divided into two groups. Nine were infected on a one-time basis (GA1) and 18 were infected on two consecutive days (GA2). Other 27 mice were inoculated with Brucella broth and divided in the same way; they composed the control group. Mice were killed at 2, 3, 7, 14 and 21 days post inoculation (pi). Fragments of stomach and duodenum were collected, fixed with 12% formalin and stained by hematoxilin-eosin and Giemsa for histopathological examination. Until the 14th()day, only reinfected mice had mild to-moderate inflammatory infiltrate in the stomach. The infiltration was predominantly lymphomonocytic, although plasma cells and eosinophils could be seen. However, at 21st day, severe eosinophilic infiltration was present in the lamina propria and submucosa of gastric corpus. In subgroup GA1, animals presented lymphomonocytic infiltration in the stomach from 14th()day pi. Our results showed that coccoid H. pylori was able to induce an acute inflammatory response in stomach of reinfected mice since the initial periods of infection. PMID- 12563491 TI - Screening and fractionation of plant extracts with antiproliferative activity on human peripheral blood mononuclear cells. AB - Three hundred and thirteen extracts from 136 Brazilian plant species belonging to 36 families were tested for their suppressive activity on phytohemaglutinin (PHA) stimulated proliferation of human peripheral blood mononuclear cells (PBMC). The proliferation was evaluated by the amount of [3H]-thymidine incorporated by the cells. Twenty extracts inhibited or strongly reduced the proliferation in a dose dependent manner at doses between 10 and 100 micro g/ml. Three of these extracts appeared to be non-toxic to lymphocytes, according to the trypan blue permeability assay and visual inspection using optical microscopy. Bioassay guided fractionation of Alomia myriadenia extract showed that myriadenolide, a labdane diterpene known to occur in this species, could account for the observed activity of the crude extract. Using a similar protocol, an active fraction of the extract from Gaylussacia brasiliensis was obtained. Analysis of the 1H and 13C NMR spectra of this fraction indicates the presence of an acetylated triterpene whose characterization is underway. The extract of Himatanthus obovatus is currently under investigation. PMID- 12563492 TI - Production of amastigotes from metacyclic trypomastigotes of Trypanosoma cruzi. AB - Attempts to recreate all the developmental stages of Trypanosoma cruzi in vitro have thus far been met with partial success. It is possible, for instance, to produce trypomastigotes in tissue culture and to obtain metacyclic trypomastigotes in axenic conditions. Even though T. cruzi amastigotes are known to differentiate from trypomastigotes and metacyclic trypomastigotes, it has only been possible to generate amastigotes in vitro from the tissue-culture-derived trypomastigotes. The factors and culture conditions required to trigger the transformation of metacyclic trypomastigotes into amastigotes are as yet undetermined. We show here that pre-incubation of metacyclic trypomastigotes in culture (MEMTAU) medium at 37 degrees C for 48 h is sufficient to commit the parasites to the transformation process. After 72 h of incubation in fresh MEMTAU medium, 90% of the metacyclic parasites differentiate into forms that are morphologically indistinguishable from normal amastigotes. SDS-PAGE, Western blot and PAABS analyses indicate that the transformation of axenic metacyclic trypomastigotes to amastigotes is associated with protein, glycoprotein and antigenic modifications. These data suggest that (a) T. cruzi amastigotes can be obtained axenically in large amounts from metacyclic trypomastigotes, and (b) the amastigotes thus obtained are morphological, biological and antigenically similar to intracellular amastigotes. Consequently, this experimental system may facilitate a direct, in vitro assessment of the mechanisms that enable T. cruzi metacyclic trypomastigotes to transform into amastigotes in the cells of mammalian hosts. PMID- 12563493 TI - Effects of chloroquine and sulfadoxine/pyrimethamine on gametocytes in patients with uncomplicated Plasmodium falciparum malaria in Colombia. AB - The effect of antimalarials on gametocytes can influence transmission and the spread of drug resistance. In order to further understand this relationship, we determined the proportion of gametocyte carriers over time post-treatment in patients with uncomplicated Plasmodium falciparum malaria who were treated with either chloroquine (CQ) or sulfadoxine/pyrimethamine (SP). The overall proportion of gametocyte carriers was high (85%) and not statistically significantly different between the CQ and SP treatment groups. However, an increased risk of carrying gametocytes on day 14 of follow up (1.26 95% CI 1.10-1.45) was found among patients having therapeutic failure to CQ compared with patients having an adequate therapeutic response. This finding confirms and extends reports of increased risk of gametocytaemia among CQ resistant P. falciparum. PMID- 12563494 TI - Clinical presentation and survival of smear-positive pulmonary tuberculosis patients of a university general hospital in a developing country. AB - From January 1995 to August 1997 we evaluated prospectively the clinical presentation, laboratory findings and short-term survival of smear-positive pulmonary tuberculosis (TB) patients who sought care at our hospital. After providing informed, written consent, the patients were interviewed and laboratory tests were performed. Information about survivorship and death was collected through September 1998. Eighty-six smear-positive pulmonary TB patients were enrolled; 26.7% were HIV-seropositive. Seventeen HIV-seronegative pulmonary TB patients (19.8%) presented chronic diseases in addition to TB. In the multiple logistic regression analysis a CD4+ cell count <= 200 cell/mm was independently associated with HIV seropositivity. In the Cox regression model, fitted to all patients, HIV seropositivity and age > or = 50 years were independently associated with decreased survival. Among HIV-seronegative persons, the presence of an additional disease increased the risk of death of almost six-fold. Use of antiretroviral drugs was associated with a lower risk of death among HIV seropositive smear-positive pulmonary TB patients (RH = 0.32, 95% CI 0.10-0.92). In our study smear-positive pulmonary TB patients had a low short-term survival rate that was strongly associated with HIV infection, age and co-morbidities. Therapy with antiretroviral drugs reduced the short-term risk of death among HIV seropositive patients after TB diagnosis. PMID- 12563496 TI - Retrenchment and restructuring in an age of austerity: what (if anything) can be learned from the affluent democracies? AB - The article discusses some difficulties of drawing implications from welfare state reform in the Organisation for Economic Co-operation and Development for "middle-income countries", and whose welfare states have been less well institutionalized. It is argued that globalization's role in contemporary social policy dynamics were not unimportant, but social processes occurring within national contexts were probably more important. There is not a single "new politics of the welfare state" but distinct political dynamics in different "regimes" or "configurations", which are characterized by different problem loads and different structures of political opportunity. Different factors are crucial in explaining outcomes in different configurations and there is little reason to expect much convergence in social policy outcomes across regime type. Contemporary welfare state reform is depicted as a process of restructuring rather than dismantling, what can encourage the possibilities for developing coalitions advancing multi-dimensional agendas of welfare state reform. PMID- 12563497 TI - [Social policy and macro economic adjustment]. AB - This paper analyzes responses by welfare states to globalization in some OECD countries in the 1980s and 90s, demonstrating that the hypothesis concerning the dismantling of social welfare systems cannot be proven due to lack of evidence of substantial changes in the funding and selection of beneficiaries. The paper also focuses on the effects of internationalization of the Brazilian economy on the country's social protection system. The conclusion is that restrictions imposed on public expenditures severely limited the capacity to promote broad coverage in social policies. During the decade the option was made for targeted, selective social policies based on the adoption of a diagnosis of low effectiveness in the Brazilian social protection system. The case of health policy was an exception, due to capacity for interest mediation in the social arena. PMID- 12563498 TI - [Equity and health sector reform in Latin America: a necessary debate]. AB - Reform and equity are terms that have frequented political discourse, technical documents, and conceptual discussions in recent decades in different proposals with different ideological references. To understand the importance and centrality of these themes in the contemporary debate in Latin America implies a more in-depth reflection on health policy in the sphere of social policies and to define which reforms were are discussing and the place equity occupies in this discussion. The first part of this essay discusses the concepts of reform and equity, followed by a review of health policy as a social policy. The article ends by discussing the central elements on the health systems reform agenda in Latin America. The conclusion is that the situation is dramatic and that recent reforms have exacerbated inequalities and created new problems by replacing the values of solidarity and equal opportunity with those of "radical utilitarian individualism"; meanwhile, the principle of "health needs" has been replaced by that of "risk", "monetarized" and defined according to the individual's social and economic position. It becomes necessary to resume the discussion of social and health policies as the matrix of principles justifying the order ascribed to any other policies. PMID- 12563499 TI - [Social inequalities and two forms of particularism in Brazilian society]. AB - Based on a discussion by contemporary social theorists concerning Brazil's historical formation and its incorporation into a project for a modern society, the article identifies two forms of particularism differences and personal relations which limit access to the market and to rights in the public sphere. The particularism of differences produces social and symbolic exclusion, hindering the formation of feelings of belonging and social interdependence, necessary for effective institutionalization of universalism in the public sphere. The particularism of personal relations permeates new institutional arrangements that incorporate participation by society in public policy management. Still, although these new arrangements and arenas bear the political and cultural marks of Brazil's historical background, they constitute open possibilities for a new equation involving universalism and particularism. PMID- 12563500 TI - [Social capital and health councils in Brazil: a virtuous circle?]. AB - This article begins by discussing the concept of social capital and its applications and limitations. Second, it examines these theoretical contributions as approaches actually used by scholars to analyze the disenchantment with the fragile political institutions and citizens' apathy that emerged together with re democratization processes in Latin America. Third, it focuses on the role of the Brazilian health councils, seeking insights from this vast and rich experience of popular mobilization and participation to shed light on the councils' contributions in fostering behaviors inherent to social capital, such as civic values and cooperation. PMID- 12563501 TI - [Racial inequalities in Brazil: a synthesis of social indicators and challenges for public policies]. AB - This article aims to systematize and analyze social data that reveal the dimension of racial inequalities in Brazil. The point of departure is that racial inequalities affect the capacity for integration of Blacks into Brazilian society and jeopardize the proposal to build a democratic society with equal opportunities for all. Such inequalities are present at different moments in the individual life cycle, beginning in childhood and continuing through school years, in access to urban infrastructure, and crystallizing in the labor market, consequently determining the income and living conditions of Afro-Brazilians as a whole. The article also analyzes the main positions in the political debate on racial inequalities in Brazil, identifying recent initiatives by both the Brazilian government and civil society to deal with racial discrimination and racial inequalities. The article concludes by identifying key challenges for health policy-makers in this context. PMID- 12563502 TI - [Family resources and educational transitions]. AB - In this paper we explore the use of a conditional logits model that allows for varying effects of socioeconomic determinants on the schooling process. The results indicate that these determinants in general have a maximum effect in the middle of transitions in primary schooling and are weaker at both the beginning and especially at the end of this schooling process. Exceptions to this pattern are the effects of "head of household's schooling", which decline over the process, and those of "per capita family income" and "skin color", which tend to increase over the course of educational transitions. PMID- 12563503 TI - [Social inequality and health in Brazil]. AB - This paper studies the relationship between social inequality and health in Brazil. The strategy adopted by the authors was to analyze needs and uses of medical care as well as access to health insurance plans according to income distribution. Determinants of health care consumption were also studied by means of logistic regression. The main source of data was the 1998 National Sample Household Survey of the Brazilian Institute of Geography and Statistics (PNAD IBGE). In general, individuals in the lowest income distribution deciles had less access to health insurance, greater need for medical care, and lower consumption of such services. Other determinants of health care consumption were heavily associated with the most privileged social strata (greater access to schooling, water supply, sewerage, electricity, garbage collection, and health insurance) and with factors pointing to the capacity to supply these services in country. PMID- 12563504 TI - [Social inequalities in the city of Rio de Janeiro: a comparison of the 1991 and 1996 censuses]. AB - This article presents a comparative model for the results of the 1996 and 1991 censuses, based on the relations indicated by the results from 1991. The authors conduct a reclassification of the social quality index (SQI) proposed by Najar (1997) for comparison of the two censuses, controlling for the variable "heads of households with 15 or more years of schooling". As is known, the 1996 census was conducted with the purpose of counting the population, and its scope of variables was quite limited. In this sense, the comparison presented here is exploratory and is limited to the variable that registers level of schooling for heads of private, permanent households, seeking to identify some changes. After situating the discussion from the perspective of analytical traditions on which it is based, the authors present and discuss the results, indicating important changes in the way the literature classically interpreted the distribution of social structure in Greater Metropolitan Rio de Janeiro. An increase was observed in years of schooling in the Rio de Janeiro population, especially among female heads of households. From the perspective of methodological tradition, this study is also in keeping with the groundbreaking work of Vetter (1981) and Pin on Charlot et al. (1986). PMID- 12563505 TI - [Poverty, inequality, and equity in health: considerations based on a transversal gender perspective]. AB - This article discusses the modernization of gender inequalities which has occurred in Brazil in the last 20 years under the hegemony of neo-liberal macro economic policies. A concept of gender as transversal is applied to questions of reproductive health (contraception and abortion, prenatal care and birthing, adolescent pregnancy, maternal and reproductive mortality, STIs/AIDS, and violence, among others), permitting analysis by both gender and social class. The history of the PAISM (Program for Integral Health Care for Women), on the other hand, reveals the complex articulation of this national public health program which, although strongly influenced by the Brazilian women's movement, has been infiltrated by heterogeneous interests in the international context. PAISM serves as an example of the appropriation of proposals and principles that were generated by this social movement, but re-articulated to gloss over the process of deepening gender and class inequality. PMID- 12563506 TI - [The challenge of health care provision politics for children and adolescents as part of equitable public policies]. AB - The author analyzes the new social policy shaped by the assimilation of the notion of integral social protection with a view towards equity. The premises marked the social reform contained in the wording of the 1988 Constitution and in the laws regulating the respective rights during the 1990s, including public social assistance, health care, and education. The article demonstrates how, in the former context, Brazil's children and adolescents were subject to great vulnerability due to their position in the country's social structure, aggravated by differential access to public goods and services. An analysis of the health care policy for children and adolescents, emphasizing an inter-sectoral approach and redefining social and health programs and measures in the 1990s, was not intended to be conclusive but did point to some trends in the reorganization of public social assistance policy for low-income youth, in keeping with gains obtained in social indicators of vulnerability in the areas of health, education, and labor during the last decade. Still, the author concludes that this policy reorientation renewed the tension between targeting more vulnerable segments of the population with selective measures as compared to universal and comprehensive access to social protection. PMID- 12563507 TI - [Combating poverty and rebuilding social ties: the lessons of Citizens' Action in the Struggle Against Hunger and Destitution and in Defense of Life]. AB - To reflect on Citizens' Action in the Struggle Against Hunger and Destitution and in Defense of Life is to seek to approach the dilemmas and challenges involving the consolidation of citizenship and social justice, as well as the forces in action in contemporary Brazilian society. However, many questions remain open in this effort. The intertwining issues of poverty, politics, and solidarity and the concrete shapes and multiple social developments of Brazil's "Campaign Against Hunger" leave room for various possible interpretations. Thus, considering the breadth of the theme on the one hand and the limits of this article on the other, the objective is to explore some relevant issues in the debate on destitution and exclusion and the process of constructing new kinds of public social intervention and civic participation, arising over the course of the study conducted by the "Citizens' Action Committees" in Rio de Janeiro during 1996 and 1997. The basic idea is to focus on volunteer practices to discuss the dilemmas posed for solving impoverishment and social fragmentation through cooperative activities and mutual help in Brazil. PMID- 12563508 TI - [Significant changes in the health system decentralization process in Brazil]. AB - This article discusses the trends and limits of the Brazilian health system decentralization process, identifying the three elements that constitute the strategic induction performed by the national system administrator in accordance with the guidelines contained in the Operational Norms of the Unified National Health System: systemic rationality, intergovernmental and service provider financing, and health care model. The effects of the Federal regulations are analyzed based on the results of the evaluation study focused on the implementation of the full management scheme at the Municipal level. The decentralization strategy induced by Basic Operational Norm 96 has succeeded in improving institutional conditions, management autonomy, and supply, as measured by the Federal resources transferred, installed capacity, production, and coverage of outpatient and hospital services, with the Municipalities authorized to conduct fully autonomous management, without altering the existing patterns of inequity in the distribution of funds to poorer Municipalities. PMID- 12563509 TI - [Health promotion and surveillance in the context of health care regionalization in the Unified National Health System in Brazil]. AB - This article discusses the limits and possibilities in the implementation of health promotion and surveillance measures, in light of the regionalization proposal contained in the Health Care Operational Norm (NOAS) adopted by the Brazilian Ministry of Health in 2001. The proposal is characterized as an opportunity to move forward in the establishment of micro-regional systems guaranteeing integral health care, in keeping with the operational capacity of the Unified National Health System in each Region and State of the country. The article goes on to analyze the contribution of various proposals for change in health practices for the development of health promotion, prevention of risks and diseases, and reorientation of individual and collective care as they are incorporated into the NOAS implementation process in various States of the country. PMID- 12563510 TI - [Brazilian Ministry of Health policy providing incentives for basic health care: a threat to the autonomy of Municipal administrators and the principle of integrality?]. AB - Thirteen years after the Unified National Health System was implemented in Brazil, the Federal government increased the use of norms and regulations aimed at rationalizing resources and prioritizing basic care within the system. In other words, although actions and services are the responsibility of Municipal governments, the Federal government used financing to reinforce its role in determining the policy to be adopted. The first part of this article analyzes trends in health care expenditures and financing in the country and priorities and strategies for financing. The second part relates the 1996 Basic Operational Norm to the Federal government policy of transferring resources to the Municipalities, discussing the importance of the Family Health Program as part of the financing logic and the role of the Health Care Operational Norm in reinforcing rationalization and emphasizing basic care. PMID- 12563511 TI - [Equity and health reforms in the 1990s]. AB - The first section of this article analyzes current issues on the agenda for research and debate in Brazilian Health Reform, from the perspective of social inclusion and exclusion. In light of the issues discussed initially, the second section analyzes the experience involving partnership between the public sector (the University Hospital at the School of Medicine, University of S o Paulo) and the Supplementary Health Care System (SSAM). The authors' hypothesis focuses on the depletion of the original set of ideals underlying the Brazilian Health Reform movement after the gains it obtained in the 1988 Constitution and in the face of the country's new reality, with the resulting need to recuperate the emphasis on the political dimension in health studies, highlighting the issue of constructing the identities of social stakeholders. Based on this initial approach, the authors proceed to analyze this partnership, demonstrating the existence of a "double waiting line" users of the Unified National Health System, or SUS, and those of the SSAM which does not imply discrimination per se in access to technology, but reproduces within the University Hospital the forms of discrimination that already exist in Brazilian society. PMID- 12563512 TI - [Delphi Study: social actors and trends in the Brazilian health care system]. AB - This article summarizes the Final Report published by the Instituto de Pesquisa Econ mica Aplicada, containing the results of the Delphi Study conducted in 2000 with the objective of identifying the main health trends in Brazil in the first decade of the 21st century, according to opinion-makers selected from various social segments most directly involved in issues in their respective sectors. A total of 138 panelists were interviewed, of whom 105 (76.1%) participated in both stages of the study (1st and 2nd questionnaires). The first major general conclusion is the improbability of any scenario involving a break with the current model. The second refers to the presence of contradictory opinions among the different groups on numerous issues. The healthy absence of a single line of thought is displayed even when all segments represented in the panel point in the same direction, since the degree of optimism (or pessimism) differs from one group to another. PMID- 12563513 TI - [Primary care policies and health reforms: an evaluative approach based on an analysis of the Family Health Program in Florianopolis, Santa Catarina, Brazil, 1994-2000]. AB - This article discusses the evaluation of primary health care policies based on a case study of the Family Health Program in Florianopolis, Santa Catarina, Brazil. The results are presented in two stages. First, the Program is placed in the context of the Municipality's policies, identifying perceptions of the Program at the administrative level. Second, the author studied the Program's practices based on a sample of five family health care teams using an evaluative framework including variables on accessibility and comprehensiveness. The Program began relatively late in the State capital, but developed a unique model. There is a greater supply of services available to the population not covered by private health plans, although there is a major diversity of practices. The degree of implementation is moderately adequate: the care provided is more comprehensive, but there are problems with access (in the ratio between staff and number of families covered). It is suggested that such proposals for reorienting health care models tend towards rationalization, political legitimization, or democratization of services. The accessibility policy can set the predominant direction. PMID- 12563514 TI - [Equity and health policy: some reflections on the Family Health Program in Brazil]. AB - This article focuses on key elements contributing to the public social policy debate, specifically from the perspective of promoting distributive justice. The reference is the Family Health Program in Brazil and its relation to recent changes in national health policy and the literature on the issues of equity and social justice. This concern is due to recent changes in social interventions by the Brazilian state, where targeting assumes a central place in the reform process. Specifically concerning health policy, the issue of equity has gained visibility, linked to the discussion on the profile of health expenditures. One of the central aspects in this context lies in the debate between notions such as the regressive and inequitable nature of targeted measures and programs, on the one hand, and the expanded perspective of access by what have traditionally been socially excluded sectors, on the other. The debate between such perspectives highlights the controversies concerning the effectiveness of health measures, equity, and the distributive concept of social justice, with the latter as the central discussion in this study. PMID- 12563515 TI - [Report. Agenda for the reduction of inequalities in health: A report from the Health and Inequality seminar: Institutions and Public Policies in the 21st Century]. PMID- 12563516 TI - Design and applications of modified oligonucleotides. AB - Oligonucleotides have a wide range of applications in fields such as biotechnology, molecular biology, diagnosis and therapy. However, the spectrum of uses can be broadened by introducing chemical modifications into their structures. The most prolific field in the search for new oligonucleotide analogs is the antisense strategy, where chemical modifications confer appropriate characteristics such as hybridization, resistance to nucleases, cellular uptake, selectivity and, basically, good pharmacokinetic and pharmacodynamic properties. Combinatorial technology is another research area where oligonucleotides and their analogs are extensively employed. Aptamers, new catalytic ribozymes and deoxyribozymes are RNA or DNA molecules individualized from a randomly synthesized library on the basis of a particular property. They are identified by repeated cycles of selection and amplification, using PCR technologies. Modified nucleotides can be introduced either during the amplification procedure or after selection. PMID- 12563517 TI - Glutamine and glutamate as vital metabolites. AB - Glucose is widely accepted as the primary nutrient for the maintenance and promotion of cell function. This metabolite leads to production of ATP, NADPH and precursors for the synthesis of macromolecules such as nucleic acids and phospholipids. We propose that, in addition to glucose, the 5-carbon amino acids glutamine and glutamate should be considered to be equally important for maintenance and promotion of cell function. The functions of glutamine/glutamate are many, i.e., they are substrates for protein synthesis, anabolic precursors for muscle growth, they regulate acid-base balance in the kidney, they are substrates for ureagenesis in the liver and for hepatic and renal gluconeogenesis, they act as an oxidative fuel for the intestine and cells of the immune system, provide inter-organ nitrogen transport, and act as precursors of neurotransmitter synthesis, of nucleotide and nucleic acid synthesis and of glutathione production. Many of these functions are interrelated with glucose metabolism. The specialized aspects of glutamine/glutamate metabolism of different glutamine-utilizing cells are discussed in the context of glucose requirements and cell function. PMID- 12563518 TI - Neuroendocrine control of body fluid homeostasis. AB - Angiotensin II and atrial natriuretic peptide (ANP) play important and opposite roles in the control of water and salt intake, with angiotensin II promoting the intake of both and ANP inhibiting the intake of both. Following blood volume expansion, baroreceptor input to the brainstem induces the release of ANP within the hypothalamus that releases oxytocin (OT) that acts on its receptors in the heart to cause the release of ANP. ANP activates guanylyl cyclase that converts guanosine triphosphate into cyclic guanosine monophosphate (cGMP). cGMP activates protein kinase G that reduces heart rate and force of contraction, decreasing cardiac output. ANP acts similarly to induce vasodilation. The intrinsic OT system in the heart and vascular system augments the effects of circulating OT to cause a rapid reduction in effective circulating blood volume. Furthermore, natriuresis is rapidly induced by the action of ANP on its tubular guanylyl cyclase receptors, resulting in the production of cGMP that closes Na+ channels. The OT released by volume expansion also acts on its tubular receptors to activate nitric oxide synthase. The nitric oxide released activates guanylyl cyclase leading to the production of cGMP that also closes Na+ channels, thereby augmenting the natriuretic effect of ANP. The natriuresis induced by cGMP finally causes blood volume to return to normal. At the same time, the ANP released acts centrally to decrease water and salt intake. PMID- 12563519 TI - Mitochondria, calcium and pro-apoptotic proteins as mediators in cell death signaling. AB - Cellular Ca2+ signals are crucial in the control of most physiological processes, cell injury and programmed cell death through the regulation of a number of Ca2+ dependent enzymes such as phospholipases, proteases, and nucleases. Mitochondria along with the endoplasmic reticulum play pivotal roles in regulating intracellular Ca2+ content. Mitochondria are endowed with multiple Ca2+ transport mechanisms by which they take up and release Ca2+ across their inner membrane. During cellular Ca2+ overload, mitochondria take up cytosolic Ca2+, which in turn induces opening of permeability transition pores and disrupts the mitochondrial membrane potential (deltapsim). The collapse of deltapsim along with the release of cytochrome c from mitochondria is followed by the activation of caspases, nuclear fragmentation and cell death. Members of the Bcl-2 family are a group of proteins that play important roles in apoptosis regulation. Members of this family appear to differentially regulate intracellular Ca2+ level. Translocation of Bax, an apoptotic signaling protein, from the cytosol to the mitochondrial membrane is another step in this apoptosis signaling pathway. PMID- 12563520 TI - Influence of the neural tube/notochord complex on MyoD expression and cellular proliferation in chicken embryos. AB - Important advances have been made in understanding the genetic processes that control skeletal muscle formation. Studies conducted on quails detected a delay in the myogenic program of animals selected for high growth rates. These studies have led to the hypothesis that a delay in myogenesis would allow somitic cells to proliferate longer and consequently increase the number of embryonic myoblasts. To test this hypothesis, recently segmented somites and part of the unsegmented paraxial mesoderm were separated from the neural tube/notochord complex in HH12 chicken embryos. In situ hybridization and competitive RT-PCR revealed that MyoD transcripts, which are responsible for myoblast determination, were absent in somites separated from neural tube/notochord (1.06 and 0.06 10(-3) attomol MyoD/1 attomol beta-actin for control and separated somites, respectively; P<0.01). However, reapproximation of these structures allowed MyoD to be expressed in somites. Cellular proliferation was analyzed by immunohistochemical detection of incorporated BrdU, a thymidine analogue. A smaller but not significant (P = 0.27) number of proliferating cells was observed in somites that had been separated from neural tube/notochord (27 and 18 for control and separated somites, respectively). These results confirm the influence of the axial structures on MyoD activation but do not support the hypothesis that in the absence of MyoD transcripts the cellular proliferation would be maintained for a longer period of time. PMID- 12563521 TI - Cissampelos sympodialis Eichl (Menispermaceae) leaf extract induces interleukin 10-dependent inhibition of Trypanosoma cruzi killing by macrophages. AB - The aqueous fraction of the ethanolic extract (AFL) of Cissampelos sympodialis Eichl (Menispermaceae), popularly known as milona, has been shown to have both immunosuppressive and anti-inflammatory effects. In the present study we investigated the modulation of macrophage antimicrobicidal activity by in vitro treatment with the extract from C. sympodialis. Normal and thioglycolate-elicited mouse peritoneal macrophages were infected in vitro with the protozoan Trypanosoma cruzi DM28c clone. We observed that the AFL (used at doses ranging from 13 to 100 microg/ml) increased T. cruzi growth and induced a 75% reduction in nitric oxide production. This inhibition could be mediated by the stimulation of macrophage interleukin-10 (IL-10) secretion since the in vitro treatment with the AFL stimulated IL-10 production by T. cruzi-infected macrophages. These results suggest that the anti-inflammatory effect of the AFL from C. sympodialis could be, at least in part, mediated by the inhibition of macrophage functions and that the inhibition of macrophage microbicidal activity induced by the C. sympodialis extract may be mediated by the decrease in macrophage function mediated by interleukin-10 production. PMID- 12563522 TI - A direct contact between astrocyte and vitreous body is possible in the rabbit eye due to discontinuities in the basement membrane of the retinal inner limiting membrane. AB - Different from most mammalian species, the optic nerve of the rabbit eye is initially formed inside the retina where myelination of the axons of the ganglion cells starts and vascularization occurs. Astrocytes are confined to these regions. The aforementioned nerve fibers known as medullated nerve fibers form two bundles that may be identified with the naked eye. The blood vessels run on the inner surface of these nerve fiber bundles (epivascularization) and, accordingly, the accompanying astrocytes lie mostly facing the vitreous body from which they are separated only by the inner limiting membrane of the retina. The arrangement of the astrocytes around blood vessels leads to the formation of structures known as glial tufts. Fragments (N = 3) or whole pieces (N = 3) of the medullated nerve fiber region of three-month-old male rabbits (Orictolagus cuniculus) were fixed in glutaraldehyde followed by osmium tetroxide, and their thin sections were examined with a transmission electron microscope. Randomly located discontinuities (up to a few micrometers long) of the basement membrane of the inner limiting membrane of the retina were observed in the glial tufts. As a consequence, a direct contact between the astrocyte plasma membrane and vitreous elements was demonstrated, making possible functional interactions such as macromolecular exchanges between this glial cell type and the components of the vitreous body. PMID- 12563523 TI - Characterization of bovine respiratory syncytial virus isolated in Brazil. AB - This paper presents the first isolation of bovine respiratory syncytial virus in Brazil and its physicochemical, morphological and molecular characterization. The virus was isolated from 33 samples of nasotracheal secretions, successively inoculated into a Madin-Darby bovine kidney cell culture, which was characterized by physicochemical tests and morphological observation by electron microscopy. The Brazilian sample is an RNA pleomorphic, enveloped, thermolabile and non hemagglutinating spicular virus. Reverse transcription, followed by nested polymerase chain reaction (nRT-PCR) assay was carried out using oligonucleotides B1, B2A, B3 and B4 for the fusion proteins (F) and B5A, B6A, B7A and B8 for the attachment protein (G). The nRT-PCR-F amplified a fragment of 481 bp corresponding to part of the gene that codes for protein F, whereas nRT-PCR-G amplified a fragment of 371 bp, in agreement with part of the G gene. The virus isolated from Brazilian samples in this study corresponded to the bovine respiratory syncytial virus, and RT-PCR proved to be useful for the diagnosis of bovine clinical samples. PMID- 12563524 TI - Absence of peripheral blood mononuclear cells priming in hemodialysis patients. AB - As a consequence of the proinflammatory environment occurring in dialytic patients, cytokine overproduction has been implicated in hemodialysis co morbidity. However, there are discrepancies among the various studies that have analyzed TNF-alpha synthesis and the presence of peripheral blood mononuclear cell (PBMC) priming in this clinical setting. We measured bioactive cytokine by the L929 cell bioassay, and evaluated PBMC TNF-alpha production by 32 hemodialysis patients (HP) and 51 controls. No difference in TNF-alpha secretion was observed between controls and HP (859 +/- 141 vs 697 +/- 130 U/10(6) cells). Lipopolysaccharide (5 microg/ml) did not induce any further TNF-alpha release, showing no PBMC priming. Paraformaldehyde-fixed HP PBMC were not cytotoxic to L929 cells, suggesting the absence of membrane-anchored TNF-alpha. Cycloheximide inhibited PBMC cytotoxicity in HP and controls, indicating lack of a PBMC TNF alpha pool, and dependence on de novo cytokine synthesis. Actinomycin D reduced TNF-alpha production in HP, but had no effect on controls. Therefore, our data imply that TNF-a production is an intrinsic activity of normal PBMC and is not altered in HP. Moreover, TNF-alpha is a product of de novo synthesis by PBMC and is not constitutively expressed on HP cell membranes. The effect of actinomycin D suggests a putative tighter control of TNF-alpha mRNA turnover in HP. This increased dependence on TNF-alpha RNA transcription in HP may reflect an adaptive response to hemodialysis stimuli. PMID- 12563525 TI - Dose-related effects of propericiazine in rats. AB - We evaluated the effects of the neuroleptic agent propericiazine on animal models of anxiety and memory. Adult male Wistar rats (250 to 350 g) received intraperitoneal injections of propericiazine (0.05, 0.075 and 0.1 mg/kg), diazepam (1 mg/kg), saline, or diazepam vehicle (20% propylene glycol and 80% saline) 30 min prior to the experimental procedure. Animals (10-15 for each task) were tested for step-down inhibitory avoidance (0.3-mA footshock) and habituation to an open-field for memory assessment, and submitted to the elevated plus-maze to evaluate the effects of propericiazine in a model of anxiety. Animals treated with 0.075 mg/kg propericiazine showed a reduction in anxiety measures (P<0.05) similar to that observed in those treated with diazepam. Propericiazine at the doses of 0.05 and 0.1 mg/kg had no significant anxiolytic effects (P>0.05) in the elevated plus-maze model of anxiety. Memory was not affected by propericiazine in any of the tests, but was impaired by diazepam. The results indicate a dose related, inverse U-shaped effect of propericiazine in an anxiety model, but not on memory tasks, perhaps reflecting involvement of the dopaminergic system in the mechanisms of anxiety. PMID- 12563526 TI - Effects of septal cholinergic lesion on rat exploratory behavior in an open field. AB - The medial septum participates in the modulation of exploratory behavior triggered by novelty. Also, selective lesions of the cholinergic component of the septohippocampal system alter the habituation of rats to an elevated plus-maze without modifying anxiety indices. We investigated the effects of the intraseptal injection of the cholinergic immunotoxin 192 IgG-saporin (SAP) on the behavior of rats in an open-field. Thirty-nine male Wistar rats (weight: 194-230 g) were divided into three groups, non-injected controls and rats injected with either saline (0.5 microl) or SAP (237.5 ng/0.5 microl). Twelve days after surgery, the animals were placed in a square open-field (120 cm) and allowed to freely explore for 5 min. After the test, the rats were killed by decapitation and the septum, hippocampus and frontal cortex were removed and assayed for acetylcholinesterase activity. SAP increased acetylcholinesterase activity in the septum, hippocampus and frontal cortex and decreased the total distance run (9.15 +/- 1.51 m) in comparison to controls (13.49 +/- 0.91 m). The time spent in the center and at the periphery was not altered by SAP but the distance run was reduced during the first and second minutes (2.43 +/- 0.36 and 1.75 +/- 0.34 m) compared to controls (4.18 +/- 0.26 and 3.14 +/- 0.25 m). SAP-treated rats showed decreased but persistent exploration throughout the session. These results suggest that septohippocampal cholinergic mechanisms contribute to at least two critical processes, one related to the motivation to explore new environments and the other to the acquisition and storage of spatial information (i.e., spatial memory). PMID- 12563527 TI - Gender-related differences in the effects of nitric oxide donors on neuroleptic induced catalepsy in mice. AB - It has been suggested that nigrostriatal dopaminergic transmission is modulated by nitric oxide (NO). Since there is evidence that gonadal hormones can affect extrapyramidal motor behavior in mammals, we investigated the effects of isosorbide dinitrate (ISD), linsidomine (SIN-1) and S-nitroso-N acetylpenicillamine (SNAP), three pharmacologically different NO donors, on neuroleptic-induced catalepsy in 60- to 80-day-old male and female albino mice. Catalepsy was induced with haloperidol (1 mg/kg, ip) and measured at 30-min intervals by means of a bar test. Drugs (or appropriate vehicle) were injected ip 30 min before haloperidol, with each animal being used only once. ISD (5, 20 and 50 mg/kg) caused a dose-dependent inhibition of catalepsy in male mice (maximal effect 120 min after haloperidol: 64% inhibition). In the females only at the highest dose of ISD was an attenuation of catalepsy observed, which was mild and short lasting. SIN-1 (10 and 50 mg/kg) did not significantly affect catalepsy in female mice, while a significant attenuation was observed in males at the dose of 50 mg/kg (maximal inhibition: 60%). SNAP (20 mg/kg) significantly attenuated catalepsy in males 120 min after haloperidol (44% inhibition), but had no significant effect on females. These results basically agree with literature data showing that NO facilitates central dopaminergic transmission, although the mechanisms are not fully understood. They also reveal the existence of gender related differences in this nitrergic modulation in mice, with females being less affected than males. PMID- 12563528 TI - Priming effects of a peripheral visual stimulus in simple and go/no-go tasks. AB - The early facilitatory effect of a peripheral spatially visual prime stimulus described in the literature for simple reaction time tasks has been usually smaller than that described for complex (go/no-go, choice) reaction time tasks. In the present study we investigated the reason for this difference. In a first and a second experiment we tested the participants in both a simple task and a go/no-go task, half of them beginning with one of these tasks and half with the other one. We observed that the prime stimulus had an early effect, inhibitory for the simple task and facilitatory for the go/no-go task, when the task was performed first. No early effect appeared when the task was performed second. In a third and a fourth experiment the participants were, respectively, tested in the simple task and in the go/no-go task for four sessions (the prime stimulus was presented in the second, third and fourth sessions). The early effects of the prime stimulus did not change across the sessions, suggesting that a habituatory process was not the cause for the disappearance of these effects in the first two experiments. Our findings are compatible with the idea that different attentional strategies are adopted in simple and complex reaction time tasks. In the former tasks the gain of automatic attention mechanisms may be adjusted to a low level and in the latter tasks, to a high level. The attentional influence of the prime stimulus may be antagonized by another influence, possibly a masking one. PMID- 12563529 TI - A brain microdialysis study on 5-HT release in freely moving rat lines selectively bred for differential 5-HT1A receptor function. AB - Breeding for high and low hypothermic responses to systemic administration of a serotonin1A (5-HT1A) receptor agonist (8-hydroxy-2-(di-n-propylamino)tetralin, 8 OH-DPAT) has resulted in high DPAT-sensitive (HDS) and low DPAT-sensitive (LDS) lines of rats, respectively. These lines also differ in several behavioral measures associated with stress. In the present microdialysis study we observed that basal 5-HT concentrations in the prefrontal cortex and dorsal hippocampus did not differ significantly between HDS and LDS rats. Thus, behavioral differences between the HDS and LDS lines might not be attributed to differences in basal 5-HT release. However, both lines had lower basal levels of 5-HT release than their randomly bred control group (random DPAT-sensitive, RDS) in the prefrontal cortex (mean +/- SEM, pg/20 microl, was 3.0 +/- 0.4 for LDS, 3.8 +/- 0.3 for HDS and 6.4 +/- 0.6 for RDS; F(2,59) = 5.8, P<0.005). The administration of (+/-)-fenfluramine (10 mg/kg) induced a greater increase in hippocampal 5-HT levels in HDS rats (500%) as compared with LDS (248%) or RDS (243%) rats (P<0.0001). There were no significant differences in the prefrontal cortex among lines, with a fenfluramine-induced 5-HT increase of about 900% in the three groups. This differential response to fenfluramine may be due to functional alterations of hippocampal 5-HT reuptake sites in the HDS line. PMID- 12563530 TI - Distribution of liposome-encapsulated antimony in dogs. AB - The achievement of complete cure in dogs with visceral leishmaniasis is currently a great challenge, since dogs are the main reservoir for the transmission of visceral leishmaniasis to humans and they respond poorly to conventional treatment with pentavalent antimonials. In order to improve the efficacy of treatment, we developed a novel formulation for meglumine antimoniate based on the encapsulation of this drug in freeze-dried liposomes (LMA). The aim of the present study was to evaluate the biodistribution of antimony (Sb) in dogs following a single intravenous bolus injection of LMA. Four healthy male mongrel dogs received LMA at 3.8 mg Sb/kg body weight and were sacrificed 3, 48 and 96 h and 7 days later. Antimony was determined in the blood, liver, spleen and bone marrow. In the bone marrow, the highest Sb concentration was observed at 3 h (2.8 microg/g wet weight) whereas in the liver and spleen it was demonstrated at 48 h (43.6 and 102.4 microg/g, respectively). In these organs, Sb concentrations decreased gradually and reached levels of 19.1 microg/g (liver), 28.1 microg/g (spleen) and 0.2 microg/g (bone marrow) after 7 days. Our data suggest that the critical organ for the treatment with LMA could be the bone marrow, since it has low Sb levels and, presumably, high rates of Sb elimination. A multiple dose treatment with LMA seems to be necessary for complete elimination of parasites from bone marrow in dogs with visceral leishmaniasis. PMID- 12563531 TI - Circadian rhythms in blood pressure in free-ranging three-toed sloths (Bradypus variegatus). AB - Blood pressure (BP) profiles were monitored in nine free-ranging sloths (Bradypus variegatus) by coupling one common carotid artery to a BP telemetry transmitter. Animals moved freely in an isolated and temperature-controlled room (24 degrees C) with 12/12-h artificial light-dark cycles and behaviors were observed during resting, eating and moving. Systolic (SBP) and diastolic (DBP) blood pressures were sampled for 1 min every 15 min for 24 h. BP rhythm over 24 h was analyzed by the cosinor method and the mesor, amplitude, acrophase and percent rhythm were calculated. A total of 764 measurements were made in the light cycle and 721 in the dark cycle. Twenty-four-hour values (mean +/- SD) were obtained for SBP (121 +/- 22 mmHg), DBP (86 +/- 17 mmHg), mean BP (MBP, 98 +/- 18 mmHg) and heart rate (73 +/- 16 bpm). The SBP, DBP and MBP were significantly higher (unpaired Student t-test) during the light period (125 +/- 21, 88 +/- 15 and 100 +/- 17 mmHg, respectively) than during the dark period (120 +/- 21, 85 +/- 17 and 97 +/- 17 mmHg, respectively) and the acrophase occurred between 16:00 and 17:45 h. This circadian variation is similar to that observed in cats, dogs and marmosets. The BP decreased during "behavioral sleep" (MBP down from 110 +/- 19 to 90 +/- 19 mmHg at 21:00 to 8:00 h). Both feeding and moving induced an increase in MBP (96 +/- 17 to 119 +/- 17 mmHg at 17:00 h and 97 +/- 19 to 105 +/- 12 mmHg at 15:00 h, respectively). The results show that conscious sloths present biphasic circadian fluctuations in BP levels, which are higher during the light period and are mainly synchronized with feeding. PMID- 12563532 TI - Different biochemical strategies of two neotropical fish to cope with the impairment of nitrogen excretion during air exposure. AB - The exposure of fish to air is normally expected to interfere with the nitrogen excretion process. Hoplias malabaricus and Hoplerythrinus unitaeniatus, two teleost species, display distinct behaviors in response to decreases in natural reservoir water levels, although they may employ similar biochemical strategies. To investigate this point, plasma levels of ammonia, urea, uric acid, and the two urea cycle enzymes, ornithine carbamoyl transferase (OCT) and arginase (ARG), as well as glutamine synthetase (GS) were determined for both species after exposure to air. Plasma ammonia increased gradually during exposure to air, but only H. malabaricus showed increased concentrations of urea. Plasma uric acid remained very low in both fish. Enzymatic activities (mean +/- SD, micromol min(-1) g protein(-1)) of H. malabaricus showed significant increases (P<0.05, N = 6) in OCT from 0.84 +/- 0.05 to 1.42 +/- 0.03, in ARG from 8.07 +/- 0.47 to 9.97 +/- 0.53 and in GS from 1.15 +/- 0.03 to 2.39 +/- 0.04. The OCT and ARG enzymes remained constant in H. unitaeniatus (N = 6), but GS increased from 1.49 +/- 0.02 to 2.06 +/- 0.03. Although these species are very closely related and share the same environment, their biochemical strategies in response to exposure to air or to increased plasma ammonia are different. PMID- 12563533 TI - Exploratory eye movements in schizophrenia: effects of figure size and the instruction on visual search. AB - It has been reported that patients with schizophrenia show restricted eye scanning in comparison with normal controls; however, the precise mechanism underlying the limited eye movement pattern remains unknown. The purpose of this study was to determine the factors affecting restricted eye-scanning in schizophrenic patients by examining exploratory eye movements during demonstration of two different sizes of the S-shaped figure. The second purpose was to determine the effect of the instruction for performance on the restricted viewing pattern in patients with schizophrenia. Eye movements during demonstration of the S-shaped figure of the original or half size were examined in 15 patients with schizophrenia and 15 normal controls using an infrared eye mark recorder. The patients showed lower search scores than control subjects for both sizes of the figure. The subjects were then instructed to compare a slightly modified figure with the original one. Lower responsive search scores were found for the patients when "fixation point" was defined as a point at which a gaze was held for at least 200 ms, while the patients and control subjects performed equally at the 100-ms setting. Direct instruction to scrutinize the S-shape abolished the difference in the search scores between patients and control subjects at both the 100-ms and 200-ms settings. These findings suggest that the size of the S-figure is not a factor of restricted eye movements, and that the direct instruction improves the visual performance in patients with schizophrenia. PMID- 12563534 TI - A prospective 2-year follow-up study of neurocognitive functioning in patients with first-episode schizophrenia. AB - To investigate the temporal stability, or progressivity, of neuropsychological (NP) impairment in schizophrenia, 50 patients with first episode (FE) schizophrenia and 50 healthy controls were given a battery of tests at the outset of the study and after a two-year interval. Both patient and control groups were balanced with respect to age, gender, education and parental socioeconomic status. Summary rating scales for semantic memory (SEM), visual memory (VIM), verbal learning (VBL), visual-motor processing and attention (VSM) and abstraction/flexibility (ABS) were constructed. FE schizophrenics showed improvement in VBL, stability of function in SEM, VSM and ABS and absence of improvement in VIM. While performance in VSM and VIM is influenced by medication status, SEM seems to be trait-related and stable; VBL, however, seems to be state related. Our data suggest that there is no proof for the assumption of progressive deterioration in NP functioning during the first few years of illness. PMID- 12563535 TI - Volumetric magnetic resonance imaging study of the anterior cingulate gyrus in schizotypal disorder. AB - Lack of normal structural asymmetry of the anterior cingulate gyrus (ACG) in patients with schizophrenia has been reported in our previous study. However, to our knowledge, no morphological studies of the brain have examined changes in ACG volume in patients with schizotypal features. We investigated the volume of the gray matter and the white matter of the ACG by three-dimensional magnetic resonance imaging (MRI) in 24 patients who met the ICD-10 criteria for schizotypal disorder (12 males, 12 females) in comparison with 48 age- and gender matched healthy control subjects (24 males, 24 females) and 40 patients with schizophrenia (20 males, 20 females). As we reported previously, right ACG gray matter volume was significantly reduced in the female patients with schizophrenia compared with the female controls. On the other hand, the gray and white matter volume of the ACG in the patients with schizotypal disorder did not differ significantly from the values in the healthy controls or the patients with schizophrenia. However, the female patients with schizotypal disorder showed a lack of right-greater-than-left asymmetry of the ACG gray and white matter found in the female controls. These results suggest that both schizotypal and schizophrenic subjects share, at least in part, the same cerebral asymmetry abnormalities. PMID- 12563536 TI - Public attitudes towards people with mental illness in six German cities: results of a public survey under special consideration of schizophrenia. AB - OBJECTIVES: Attitudes of the urban population in Germany towards people with mental illness were investigated in this study. The results are compared with those of attitude surveys conducted by other research centres participating in the World Psychiatric Association's (WPA) global anti-stigma-programme "Fighting Stigma and Discrimination because of Schizophrenia - Open the Doors" (WPA 1998). METHODS: A total of 7246 German-speaking persons aged 16 and over were interviewed in private households in six German cities by telephone using a standardised questionnaire. The respondents were asked about their knowledge in regard to schizophrenia, their social distance towards people with schizophrenia and estimations of the social stigmatisation of mental patients in general. RESULTS: 33.1 % of the interviewees were able to name causes of schizophrenia. 76.5 % of the interviewees believe that people with schizophrenia often or very often need prescription drugs to control their symptoms. 81.1 % believe that most people would pass over the job application of a former mental patient in favour of another applicant. CONCLUSIONS: Improvements in the education of the public about mental illnesses and provision of the opportunity for personal contact with mentally ill people are considered to be important measures for promoting the acceptance of the mentally ill by the public. PMID- 12563537 TI - Can only reversed vegetative symptoms define atypical depression? AB - BACKGROUND: The definition of atypical depression (AD) has recently seen a rebirth of studies, as the evidence supporting the current DSM-IV atypical features criteria is weak. Study aim was to compare a definition of AD requiring only oversleeping and overeating (reversed vegetative symptoms) to the DSM-IV AD definition (always requiring mood reactivity, plus overeating/weight gain, oversleeping, leaden paralysis, and interpersonal sensitivity [at least 2]). METHODS: Consecutive 202 major depressive disorder (MDD) and 281 bipolar II outpatients were interviewed, during a major depressive episode (MDE), with the Structured Clinical Interview for DSM-IV. The DSM-IV criteria for AD were compared to a new AD definition based only on oversleeping and overeating, which was the one often used in community studies. Associations were tested by univariate logistic regression. RESULTS: The frequency of DSM-IV AD was 42.8 %, and that of the new AD definition was 38.7 %. DSM-IV AD, and the new AD definition, had almost all the same significant associations: bipolar II, female gender, lower age, lower age of onset, axis I comorbidity, depressive mixed state, MDE symptoms lasting more than 2 years, and bipolar family history. DSM-IV AD was present in 86 % of the new AD definition sample. The new definition of AD was significantly associated with all the other DSM-IV AD symptoms not included in it. The new AD definition was strongly associated with DSM-IV AD (odds ratio = 17.8), and had sensitivity = 77.7 %, specificity = 90.5 %, positive predictive value = 86.1 %, negative predictive value = 84.4 %, and ROC area curve = 0.85, for predicting DSM-IV AD. CONCLUSIONS: Results support a simpler definition of AD, requiring only oversleeping and overeating, and support the similar AD definition previously used in community studies. This definition is easier and quicker to assess by clinicians than the DSM-IV definition (mood reactivity and interpersonal sensitivity are more difficult to assess). Some pharmacological studies support this new AD definition (by showing better response to MAOI than to TCA, as shown in DSM-IV AD). PMID- 12563538 TI - Weight gain and glucose and lipid metabolism disturbances during antipsychotic medication: a review. AB - Antipsychotic medication is the mainstay of treatment for functional psychotic illnesses. However, for some patients weight gain and the disturbances in blood lipid levels and glucose balance associated with their use are significant disadvantages, and pose health risks that may affect prognosis. A substantial body of evidence suggests that weight gain is at least partly related to the blocking effects of antipsychotic medication on serotonin- and histamine-mediated neurotransmission. The disadvantages associated with weight gain can be reduced by an appropriate choice of antipsychotic and avoidance of polypharmacy, by regular monitoring of the patient's weight, and, if necessary, by the patient's participation in a dieting programme. PMID- 12563539 TI - Is hemispheric language dominance relevant in musical hallucinations? Two case reports. AB - It has been suggested that musical hallucinations are often due to a focal brain lesion predominantly of right hemispheric pathologies. We report about two patients with musical hallucinations associated with a brain tumor of the right hemisphere. Although both patients were right-handed, one of them had a right hemispheric dominance for language as proven by the Wada test. The musical ability as measured by the Seashore test was not impaired. We conclude that the language dominance of the hemisphere is not primarily relevant for the development of musical hallucinations. PMID- 12563540 TI - Sphingosylphosphorylcholine stimulates cellular fibronectin expression through upregulation of IL-6 in cultured human dermal fibroblasts. AB - Sphingosylphosphorylcholine (SPC) has been reported to stimulate wound healing by its potent mitogenic effect. Fibronectin (FN) is a cell-adhesion protein that plays an important role in cell migration and collagen deposition during wound healing. In order to elucidate further the mechanism involved in the accelerated wound healing stimulated by SPC, we studied the role of SPC in FN production in cultured human dermal fibroblasts. We demonstrated that SPC dose- and time dependently enhanced the expression of FN in human dermal fibroblast at the protein and mRNA levels. IL-6 is known to stimulate the production of FN in fibroblasts. SPC also markedly induced IL-6 production in cultured human dermal fibroblasts in a dose- and time-dependent manner. We also demonstrated that FN mRNA expression in human dermal fibroblasts was upregulated 4 h after IL-6 treatment. Moreover, pretreatment with neutralizing anti-IL-6 antibodies partially blocked the upregulation of FN mRNA expression induced by SPC in human dermal fibroblasts. These results indicate that SPC may stimulate FN synthesis through IL-6 production in cultured human dermal fibroblasts. PMID- 12563541 TI - A pitfall in clinical photography: the appearance of skin lesions depends upon the illumination device. AB - BACKGROUND: Pictures of skin lesions are usually recorded with a camera equipped with a ring-type electric flash. In some cases the photographs do not reproduce the image perceived by the naked eye. OBJECTIVE: To investigate the quality of photographs taken with the ring-type electric flash. METHODS: Photographs of skin lesions and an experimental skin model were taken under different illumination conditions including a standard light source, full-spectrum fluorescent ceiling lights and the ring-type electric flash. RESULTS: Images taken under illumination with a standard light source box resembled those perceived by the naked eye. The full-spectrum fluorescent ceiling lights produced adequate results. Pictures taken with the ring-type electric flash showed some deficiencies including reduction in contrast, loss of shading and excess reflection. CONCLUSIONS: The ring-type electric flash is not an ideal device for recording skin lesions. An illumination device compatible with a standard light source should be employed to yield maximum fidelity. PMID- 12563542 TI - Fibroblasts play a stimulatory role in keratinocyte proliferation but an inhibitory role in melanocyte growth and pigmentation in a skin equivalent system from skin type IV. PMID- 12563543 TI - Helicobacter pylori not detected in cutaneous mucosa-associated lymphoid tissue (MALT) lymphomas. PMID- 12563544 TI - Pharmacological characterization of the effects of 3,4 methylenedioxymethamphetamine ("ecstasy") and its enantiomers on lethality, core temperature, and locomotor activity in singly housed and crowded mice. AB - RATIONALE: Few studies have directly compared the effects of methylenedioxymethamphetamine (MDMA, "ecstasy") and its enantiomers across measures. OBJECTIVES: To investigate the capacity of MDMA and its stereoisomers to produce aggregate toxicity in mice, the influence of 5-HT(2) receptors, 5-HT transporters, and ambient temperature on this effect, and to directly compare the racemate and its enantiomers in terms of their effects on core temperature and locomotor activity with and without various serotonergic pretreatments. METHODS: Mice were injected with various doses of MDMA and its stereoisomers in various housing conditions, with and without pretreatments of serotonergic drugs, and at two distinct ambient temperatures; lethality was quantified 2 h after MDMA administration. For temperature/activity studies, mice were injected with various doses of MDMA and its enantiomers, with and without ketanserin, MDL100907, or fluoxetine pretreatments, and core temperature and locomotor activity data were collected for 24 h. RESULTS: Racemic MDMA and its isomers produced aggregate toxicity in mice. The lethal effects of racemic MDMA and its enantiomers were differentially attenuated by the various serotonergic pretreatments and manipulation of the ambient temperature across housing conditions. Racemic and S(+)-MDMA produced hyperthermic effects in mice, while R(-)-MDMA did not. The pretreatment drugs attenuated the hyperthermic effects of racemic MDMA, but were less effective in blocking S(+)-MDMA-induced hyperthermia. Racemic MDMA and both enantiomers stimulated locomotor activity, although R(-)-MDMA was least effective. The pretreatments all reduced the locomotor stimulant effects of racemic MDMA but potentiated S(+)-MDMA-induced hyperlocomotion. CONCLUSIONS: The MDMA isomers have heterogeneous effects that can be demonstrated across a wide range of endpoints. PMID- 12563545 TI - Allelic variation in the serotonin transporter promoter affects neuromodulatory effects of a selective serotonin transporter reuptake inhibitor (SSRI). AB - RATIONALE: Antidepressant efficacy of selective serotonin reuptake inhibitors (SSRIs) has been shown to depend on functional polymorphisms within the promoter region of the serotonin transporter gene (5-HTTLPR). This gene gives rise to a biallelic polymorphism designated long (l) and short (s). Homozygosity for the long variant (ll-genotype) is associated with a two times more efficient 5-HT uptake compared to the s/l- or s/s-genotype. Paired pulse transcranial magnetic stimulation is a feasible tool in detecting changes of motor cortex excitability induced by SSRIs. OBJECTIVE: Our study aimed to measure neuromodulatory effects of SSRIs on cortical excitability in healthy volunteers characterized by distinct genotypes of the 5-HTTLPR. METHODS: Cortical excitability was determined in eight genetically defined subjects pre- and post-ingestion of 60 mg citalopram. RESULTS: Subjects with the ll-genotype of the 5-HTTLPR showed a significant enhancement of a particular component of motor cortex excitability (intracortical inhibition) as compared to volunteers without the ll-genotype. CONCLUSION: Distinct neuromodulatory effects after intake of citalopram based on allelic variations of the 5-HTTLPR may explain variable response of patients treated with SSRIs. PMID- 12563546 TI - The use of multiple informants in identifying the risk factors of depressive and disruptive disorders--are they interchangeable? AB - OBJECTIVE: The aim of this study was to asses whether children and their parents identify the same risk factors for disruptive and depressive disorders and to analyze whether combining informant data with a rule that classifies the diagnosis as present if confirmed by at least one informant (OR rule) masks distinctive patterns identified in informant-specific analyses. METHOD: Bivariate logistic regression equations were estimated using the diagnostic classification, based on DISC 2.1, as the outcome variable and informant (parent or child), characteristics of the youth (gender and age), indicators of the context of the interview (site), and family characteristics (income, parental monitoring, and adverse family environment) as predictors. The same predictors were also analyzed with the combined informant (OR rule) as outcome variable. RESULTS: Prevalence of all diagnoses varied with informant. Depressive disorders were more prevalent when the informant was the youth and disruptive disorders when the informant was the parent. The effect of age varied with informant. Odds of being classified as having a DISC disorder increased with age when the informant was the youth but the same effect was not observed when the parent was the informant. When information from parents and youth are combined (with an OR rule) the age effect for disruptive disorders vanishes, and its effect for depressive disorders weakens. CONCLUSIONS: Informants are not interchangeable. Parent- and youth-based estimates of the prevalence of disruptive and depressive disorders were different and showed distinctive age relationships. Combining information from different sources (parents and youths) obscures the apparent effect of age noted in the two informant groups. PMID- 12563547 TI - Psychological distress and socioeconomic status in single mothers and their children in a German city. AB - BACKGROUND: The proportion of single mothers in western countries is continuously growing. In contrast to other western countries, support programs in Germany especially for single mothers are rare. This study identifies for the first time in Germany within a large epidemiological sample different subgroups of higher distressed single mothers as important potential target groups for psychosocial support interventions. Facing limited resources, data about these subgroups are helpful to develop and establish specific support programs. METHOD: In a cross section study, family status of a complete cohort of school beginners in Duesseldorf (N = 5178 children, aged 5-7 years) was screened within the school eligibility test (recruitment rate 97.5 %, N = 5048). Out of a total of 891 single mothers, 531 fulfilling inclusion criteria (speaking fluent German, not married, living together with the tested child, informed consent) were compared to a control group of married mothers out of the same sample (N = 278). Socioeconomic status, social network and psychological distress of mothers (SCL 90-R) and children (CBCL) were investigated. RESULTS: Socioeconomic status (income, education) of single mothers was lower while psychological distress (SCL 90-R) was elevated compared to the control group. Single mothers without additional personal support for their child, younger, as well as poor single mothers showed higher values of psychological distress. In sons of single mothers, increased behavior problems were found (CBCL). Of all children screened, 907 (18 %) lived in single-parent families. CONCLUSION: An increased psychological distress of single mothers and their sons could be shown for the first time in Germany within a large epidemiologic sample. This has implications for the planning of preventive interventions and evaluation of associations, e. g., between social variables and distress of single mothers and their children within longitudinal study designs. PMID- 12563548 TI - Effectiveness of psychoeducational intervention for rural Chinese families experiencing schizophrenia--a randomised controlled trial. AB - BACKGROUND: The aim of this study was to explore the characteristics and efficacy of psychoeducational family intervention for persons with schizophrenia in rural China. METHODS: A cluster randomised controlled trial of psychoeducational family intervention for families experiencing schizophrenia (three groups, 326 cases) was conducted in Xinjin County, Chengdu. Treatment groups consisted of family intervention and medication, medication alone, and a control. RESULTS: The results showed a gain in knowledge, a change in the relatives' caring attitudes towards the patients, and an increase in treatment compliance in the psychoeducational family intervention group (p < 0.05, 0.001). Most importantly, the relapse rate over 9 months in this group (16.3 %) was half that of the drug only group (37.8 %), and just over one-quarter of that of the control group (61.5 %) (p < 0.05). Antipsychotic drug treatment and families' attitudes towards patients after the 9-month follow-up were significantly associated with clinical outcome (p < 0.05). CONCLUSIONS: In rural China, family intervention should focus on improving the relatives' recognition of illness, the caring attitude towards the patients, treatment compliance, relapse prevention, and the training of the patients' social functioning. This trial, one of the largest in the literature, has shown that psychoeducational family intervention is effective and suitable for psychiatric rehabilitation in Chinese rural communities. PMID- 12563549 TI - Predicting vocational functioning and outcome in schizophrenia outpatients attending a vocational rehabilitation program. AB - BACKGROUND: Vocational rehabilitation is a central issue in the rehabilitation of patients with chronic schizophrenia. However, even with the help of comprehensive integration programs, achieving this objective remains a very ambitious and difficult undertaking. Therefore, a profound and up-to-date knowledge of vocational functioning and outcome predictors in patients who have the goal to return into competitive employment is imperative. The objective of the present study was to test the predictors summarized in the recent review of Cook and Razzano, as well as to test those predictors specified in the nine hypotheses put forward by Anthony and Jansen in schizophrenia patients enrolled in a vocational rehabilitation program. METHODS: The predictive value of ten hypotheses centering on vocational functioning and outcome were consecutively tested in a sample of 53 schizophrenia patients. Those predictors identified as significant were then taken into a 'winner take all' regression in order to determine which of them were the best. RESULTS: The overall work performance observed in a workshop proved to be the best predictor of vocational functioning. Contrary to the pivotal claim in Anthony and Jansen's review, in our sample, negative symptoms indeed influenced vocational functioning, outcome and functional skills. Cognitive impairments, social competence and fatalistic control beliefs also had predictive value for vocational functioning and outcome. CONCLUSIONS: These results reflect the accumulated findings of the past decade as summarized by Cook and Razzano. Moreover, they serve to substantiate the necessity of promoting the concept of cognitive remediation and associated programs designed to transform fatalistic beliefs into feelings of hopefulness, thereby enhancing the readiness of schizophrenia patients to enroll in rehabilitation programs. PMID- 12563550 TI - Predictors of improvement in quality of life of severely mentally ill substance abusers during 18 months of co-operation between psychiatric and social services. AB - BACKGROUND: The aim of this study was to investigate predictors of Quality of Life in a group of severely mentally ill substance abusers. These patients took part in a multi-centre study aimed at improving co-operation between psychiatric and social services in Sweden during the years 1995 to 1998. METHODS: Two hundred and eighty-eight patients, 62.4 % men, were included in the study. The criteria to enter the study were to have a diagnosis of severe mental illness and a diagnosis of substance dependence according to the DSM-III-R criteria. Quality of Life (QoL) was measured by a global assessment, Cantril's ladder (1965). Initially and after 18 months the following measurements were also used: Addiction Severity Index (ASI), Symptom Check List 90 (SCL-90) and The Clinical Rating Scale (CRS) for Alcohol Use (AUS) and Drug Use (DUS). RESULTS: Initially those who were older and those who had an apartment of their own or who lived in sheltered living had a higher QoL than the others. Those belonging to the borderline personality disorder subgroup had a lower QoL than those belonging to other psychiatric diagnostic subgroups. At follow-up QoL had improved significantly. Improvement in QoL was related to improvements in physical health, legal and family problems, psychiatric symptoms and a reduction of alcohol and drug problems (ASI), global functioning (GAF) and psychological problems (SCL 90). A multiple stepwise regression analysis showed that improvement in QoL primarily was predicted by improvements in psychiatric symptoms. Number of months without alcohol and drugs were positively associated with improvement in QoL. As a whole, at follow-up the QoL is still not high. CONCLUSIONS: In this group of severely mentally ill substance abusers, improvement in QoL was primarily predicted by improvements in psychiatric symptoms. Further, less alcohol and drug abuse seems to augment the subjective feeling of QoL. PMID- 12563551 TI - Sociodemographic and clinical characteristics of patients with conversion disorder in Eastern Turkey. AB - BACKGROUND: Conversion disorder (CD) is a common disease and its importance still continues in Turkey and particularly in Eastern Turkey. The aim of this study is to examine sociodemographic and clinical characteristics of CD. METHOD: Among 198 consecutive patients having CD diagnosed by structured DSM-III-R clinical interview, the psychosocial characteristics of the patients were clinically investigated. RESULTS: The most common subtype of CD was non-epileptic seizure (NES) (41.4 %). The psychosocial stress factors were found in the initiation or at the last episode of the disorder (88.9 %). The most prominent problem related with primary support group was traumatic event (37.9 %) followed by problems associated with migration and related economical problems which are the most important problems of the study area. The incidence of depressive disorders was high in patients with CD, and the histrionic personality disorder was the most prominent personality pathology among the patients. Direct referral to psychiatry clinics appeared to be low (12.1 %). CONCLUSION: Our findings have shown that traumatic events may have an important role in the occurrence, severity and duration of CD, and most of the patients seek help from religious healers. The study has also revealed that lower education level and socioeconomic and sociocultural problems may play a role in the occurrence of the disorder as well as regarding its course. PMID- 12563552 TI - The development of the Chinese version of Impact of Event Scale--Revised (CIES R). AB - BACKGROUND: This is an exploratory study on the psychometric properties and normative data of the Chinese translation of the Impact of Event Scale - Revised (CIES-R). METHODS: The original Impact of Event Scale - Revised was translated into Chinese and the comparability of content was verified through back translation procedures. The study consisted of a sample of 116 patients from the Accident and Emergency Department. RESULTS: The reliability of the Chinese version was verified. The validity of the Chinese version as a measure of psychological distress was supported by the positive correlations between the various subscale scores of CIES-R (i. e., Intrusion, Avoidance, and Hyperarousal) and General Health Questionnaire-20. However, the independence of the three subscales was not supported. CONCLUSIONS: The present result suggests that normative information for different age groups and clinical samples might be different. Further study with clinical sample and investigation of the relationship between CIES-R and other diagnostic measures of PTSD are discussed. PMID- 12563553 TI - Temporal sequencing of lifetime mood disorders in relation to comorbid anxiety and substance use disorders--findings from the Netherlands Mental Health Survey and Incidence Study. AB - BACKGROUND: Little is known about the temporal sequencing of psychiatric disorders. The aim of this study was to obtain insight into patterns of co occurrence of DSM-III-R mood disorders in relation to anxiety and substance use disorders, their temporal sequencing and the sociodemographic and long-term vulnerability predictors of this temporal sequencing. METHODS: Data are from the Netherlands Mental Health Survey and Incidence Study (NEMESIS), a psychiatric epidemiological study in a representative sample of 7,076 adults aged 18-64. RESULTS: Of those who had ever experienced a mood disorder, 46 % of males and 57 % of females had a history of anxiety disorders, and 43 % and 15 % of substance use disorders. Mood disorders were associated with all anxiety and substance use disorders, except with alcohol abuse among males. In the majority of anxiety comorbid cases, the mood disorder arose after the anxiety disorder; the pattern for substance use-comorbid disorders was more variable. Deviation from the usual sequence of major depression and anxiety disorders was more often seen among females, subjects with a higher educational level, subjects who experienced childhood parental divorce, and subjects who experienced childhood emotional neglect. CONCLUSIONS: When comorbid with anxiety disorders, mood disorders clearly tend to be secondary. Few of the studied demographic factors, familial vulnerability factors and childhood life events predict the sequencing of mood disorders in relation to other disorders. PMID- 12563554 TI - Gender differences in the association of adult hopelessness with adverse childhood experiences. AB - BACKGROUND: The impact of childhood traumatic events on long-term psychological development has been widely studied. Nevertheless, little research has been carried out on possible associations between adverse childhood experiences (ACEs) and hopelessness in adulthood, and whether any gender differences exist. AIM: The aim of this study was to examine the association between ACEs (poor relationship between parents, unhappiness of childhood home, hard parenting, physical punishment, domestic violence, alcohol abuse in primary family) and current hopelessness without any mental disorder in a general population sample. METHOD: 1598 adults (43 % were men), aged 25-64 years, completed self-report measures to assess ACEs and hopelessness by means of the Beck Hopelessness Scale (HS). Logistic regression was used to adjust for the effects of sociodemographic factors on the association between the cumulative number of ACEs and hopelessness. RESULTS: Whereas several bivariate associations were found between ACEs and hopelessness, none of them remained significant in multivariate analysis. However, men who reported three or more ACEs were 2.79 times (95 % CI 1.17-6.63) and women 2.19 times (95 % CI 1.04-4.65) more likely to be hopeless compared with those without any ACEs. In women (OR 2.25, 95 % CI 1.01-5.00), but not in men, this relationship remained significant after adjusting for several current covariates. CONCLUSION: Clustering of ACEs may have long-lasting effects by increasing the risk of hopelessness in adulthood, especially in women. Increased awareness of the frequency of ACEs and their subsequent consequences, such as hopelessness, may encourage health care professionals to undertake preventive work in primary and mental health care. PMID- 12563555 TI - Self-reported suicidal and help seeking behaviours in the general population in Latvia. AB - BACKGROUND: The aims of this study were to assess the prevalence of suicidal behaviours in the general population in Latvia, to identify risk groups for suicidal behaviour, to examine a possible continuous sequence of suicidal behaviours with underlying gradient of severity, to assess patterns of help seeking behaviours and preferences of different types of services for suicidal persons. METHOD: A postal survey of a stratified proportional sample of the general population aged 18 and older was carried out. RESULTS: The last year and lifetime prevalence of any type of suicidal behaviours was 52.6 and 60.2 %. The incidence of self-reported suicide attempts was 1.8 and 5.1 %, respectively. Females reported significantly less serious types (includes ideation, plans and/or attempts) of suicidal behaviours (OR 0.04, p = 0.001) during last year than males. Younger age, lower level of education, urban residency and Latvian ethnicity were identified as risk factors for serious types of suicidal behaviours in both genders. Non-cohabitation status (OR 5.3, p = 0.01) and lower level of education for males, but higher levels of education for females were identified as significant risk factors for mild types (solely life-weariness and/or death wishes) of suicidal behaviours. The results indicated no simple cumulative relationship between the suicidal behaviours. Age, gender, previous help seeking experience and severity of self-reported suicidal behaviours influenced help seeking behaviours. The overall acceptance of professional services was high, but people who reported suicidal behaviours ranked them lower. CONCLUSIONS: A postal survey can provide a reasonable coverage of suicidal behaviours and description of risk groups in the general population. Higher prevalence of suicidal behaviours among males might indicate that nowadays males are under certain stress in Latvia. PMID- 12563556 TI - Schizophrenia: illness impact on family members in a traditional society--rural Ethiopia. AB - BACKGROUND: Studies have consistently shown that both the subjective and objective dimensions of burden among family members of schizophrenia patients and other psychiatric disorders are prevalent. However, as most of these reports were from western societies, we lack information on the subject in developing countries. METHOD: The study was conducted within the framework of the ongoing epidemiological study of course and outcome of schizophrenia and bipolar disorders in a rural population of 15-49 years of age. Three hundred and one cases of schizophrenia and their close relatives participated in the study. RESULTS: Family burden is a common problem of relatives of cases with schizophrenia. Financial difficulty is the most frequently endorsed problem among the family burden domains (74.4 %). Relatives of female cases suffered significantly higher social burden (Z = 2.103; p = 0.036). Work (Z = 2.180; p = 0.029) and financial (Z = 2.088; p = 0.037) burdens affected female relatives more often than males. Disorganised symptoms were the most important factors affecting the family members in all family burden domains. Prayer was found to be the most frequently used coping strategy in work burden (adj. OR = 1.99; 95 % CI = 1.08-3.67; p = 0.026). CONCLUSION: Negative impact of schizophrenia on family members is substantial even in traditional societies such as those in Ethiopia where family network is strong and important. The scarce existing services in the developing countries should include family interventions and support at least in the form of educating the family members about the nature of schizophrenia illness and dealing with its stigma and family burden. PMID- 12563557 TI - Traumatic events, migration characteristics and psychiatric symptoms among Somali refugees--preliminary communication. AB - BACKGROUND: Each refugee group experiences specific migration and resettlement experiences. There are no epidemiological data on risk factors for psychiatric symptoms among adult Somalis in the UK. METHODS: We interviewed a community sample of 180 Somalis. We assessed the relationship between symptoms of psychosis (BPRS), anxiety and depression (SCL-90) and suicidal thinking (BDI) and migration related experiences such as traumatic events, immigration difficulties, employment and income. RESULTS: Anxiety and depression was incrementally more common with each pre-migration traumatic event (OR per trauma event = 1.31, 1.06 1.62, p = 0.01). Shortages of food, being lost in a war situation, and being close to death and suffering serious injury were each related to specific psychiatric symptoms. Suicidal thinking was more common among Somalis who were unemployed before migration and those using qat in the UK. CONCLUSIONS: War related experiences, occupational status before migration and current Qat use are risk factors for psychiatric symptoms among Somali refugees. PMID- 12563558 TI - Interventions and outcomes of health and social care service provision for people with severe mental illness in England. AB - BACKGROUND: Mental health policy in England is undergoing radical change involving the integration of services aimed at improving outcomes for patients. At the same time, there is limited evidence about how conventional services are performing. The present paper reports data on the services provided and short term outcomes achieved in eight community services in England. METHOD: A survey of caseloads of nurses and social workers was undertaken using a single-page assessment tool (MARC1) (n = 3024). After 5 months a random sample of psychotic cases was followed up (n = 393). RESULTS: A tendency was observed for health and social care practitioners to use the services available from within their own organisation. Over time, in the most severe cases, there was a substantial increase in provision of the services of the other organisation. Outcomes in terms of changes in HoNOS, GAS and MARC1 scores were similar for both professional groups, and both reported similar amounts of met and unmet need (and in the same categories) at follow-up. CONCLUSION: The most likely explanation for the change in service provision is the separate operation of different professional groups acting as gatekeepers for their own resources. PMID- 12563559 TI - [Achilles tendon ruptures: 25 year's experience in sport-orthopedic treatment]. AB - From 1972 - 1996 570 Achilles tendon ruptures in 565 patients were treated in the Sportklinik Stuttgart. The 499 men and 66 women had an average age of 38 years. For the diagnosis of a Achilles tendon rupture Ultrasound and MRI are important procedures, but clinical history and examination are still the best methods to find an Achilles tendon rupture (100%). However,the Actiology of the Achilles tendon rupture is still controversial and cannot be answered by these methods. Opposed to the degenerative theory, biomechanical experiments show that any Achilles tendon can tear when the calf muscle is tensed before the tendon is quickly stretched. We found that 69.8% of the patients with Achilles tendon rupture had a real trauma. Regardless of that, the treatment of the ruptured Achilles tendon has considerably changed over the last ten years. Responsible for this development are the positive experiences at the field of sports medicine with minimally invasive methods and the early functional treatment after knee surgery. Since we use an early functional rehabilitation concept instead of plaster immobilisation, all methods to treat a ruptured Achilles tendon have been improved. 43.5% of the patients after plaster immobilisation and 28.8% of the patients after early functional rehabilitation had a subjectively felt force reduction. Other important selecting criteria are the risk factors related to treatment method. Minimal invasive percutaneous Achilles tendon repair is considerably better than conservative therapy with a high rate of re-rupture (9.8%) and better than the open surgical repair, which carries a higher risk of infection (2.2%) PMID- 12563560 TI - Confirmation of linkage of prostate cancer aggressiveness with chromosome 19q. AB - Regions on chromosomes 7 and 19 were recently reported to contain susceptibility loci that regulate tumor aggressiveness of prostate cancer. To confirm these findings, we analyzed genome scan data from 161 pedigrees affected with prostate cancer. Using the Gleason score as a quantitative measure of tumor aggressiveness, we regressed the squared trait difference, as well as the mean corrected cross product, on the estimated proportion of alleles shared identical by-descent at each marker position. Our results confirm the previous linkage results for chromosome 19q (D19S902, P<.00001). In addition, we report suggestive evidence for linkage on chromosome 4 (D4S403, P=.00012). The results of previous findings, together with our results, provide strong evidence that chromosome 19 harbors a gene for tumor aggressiveness. PMID- 12563561 TI - Mapping the Wolf-Hirschhorn syndrome phenotype outside the currently accepted WHS critical region and defining a new critical region, WHSCR-2. AB - In an attempt to define the distinctive Wolf-Hirschhorn syndrome (WHS) phenotype, and to map its specific clinical manifestations, a total of eight patients carrying a 4p16.3 microdeletion were analyzed for their clinical phenotype and their respective genotypes. The extent of each individual deletion was established by fluorescence in situ hybridization, with a cosmid contig spanning the genomic region from MSX1 (distal half of 4p16.1) to the subtelomeric locus D4S3359. The deletions were 1.9-3.5 Mb, and all were terminal. All the patients presented with a mild phenotype, in which major malformations were usually absent. It is worth noting that head circumference was normal for height in two patients (those with the smallest deletions [1.9 and 2.2 Mb]). The currently accepted WHS critical region (WHSCR) was fully preserved in the patient with the 1.9-Mb deletion, in spite of a typical WHS phenotype. The deletion in this patient spanned the chromosome region from D4S3327 (190 b4 cosmid clone included) to the telomere. From a clinical point of view, the distinctive WHS phenotype is defined by the presence of typical facial appearance, mental retardation, growth delay, congenital hypotonia, and seizures. These signs represent the minimal diagnostic criteria for WHS. This basic phenotype maps distal to the currently accepted WHSCR. Here, we propose a new critical region for WHS, and we refer to this region as "WHSCR-2." It falls within a 300-600-kb interval in 4p16.3, between the loci D4S3327 and D4S98-D4S168. Among the candidate genes already described for WHS, LETM1 (leucine zipper/EF-hand-containing transmembrane) is likely to be pathogenetically involved in seizures. On the basis of genotype phenotype correlation analysis, dividing the WHS phenotype into two distinct clinical entities, a "classical" and a "mild" form, is recommended for the purpose of proper genetic counseling. PMID- 12563562 TI - Regression of left ventricular hypertrophy: hoping for a longer life. AB - Left ventricular hypertrophy (LVH) is more than just an adaptive response to hypertension. It predicts a poor prognosis independently of the blood pressure (BP) level. There is increasing evidence from studies such as Heart Outcomes Prevention Evaluation (HOPE) and Losartan Intervention For Endpoint reduction in hypertension (LIFE) that LVH should be a target for treatment, above and beyond BP control. It is likely that drugs blocking the renin-angiotensin-aldosterone system cause greater regression of LVH than other agents and this is probably the mechanism that explains the superiority of losartan over atenolol in the LIFE study. In order to achieve the stringent BP goals suggested by modern guidelines, most patients will require multiple antihypertensive agents and the clinical choices relate more often to which combinations of drugs are most appropriate, rather than which single drug is the best. Tight BP control should be the first priority and this is likely to lead to regression of LVH. PMID- 12563563 TI - Developments in restenosis. AB - Restenosis is a major complication leading to the failure of vascular procedures, including surgery, angioplasty and stenting. Major efforts including over 100 clinical trials have been made to overcome this complication, with little success to date. Issues relating to trial rationale, design, measurement and biology are addressed in this review. PMID- 12563564 TI - The potential benefits of aldosterone antagonism in Type 2 diabetes mellitus. AB - Interest in the renin-angiotensin-aldosterone system (RAAS) has increased since the development of angiotensin-converting enzyme (ACE) inhibitors. It has been discovered that the potential uses of this class of treatment extend far beyond their initial developmental role as antihypertensives, and they are now used routinely in the treatment of heart failure, nephropathy, myocardial infarction and diabetes. However, there is more to RAAS blockade than just inhibition of angiotensin II, and inhibition of aldosterone is becoming recognised as an additional therapeutic manoeuvre in chronic heart failure. Since inhibition of the RAAS at the level of ACE is now seen to be an important therapy in diabetes; the purpose of this article is to explore the potential benefits of additional aldosterone inhibition in Type 2 diabetes mellitus. PMID- 12563565 TI - Vasopeptidase inhibitors in heart failure. AB - Considerable attention has recently focused on the vasopeptidase inhibitors (VPI), a new class of drug that combines angiotensin-converting enzyme (ACE) inhibitor activity with inhibition of natriuretic peptide breakdown. In theory, a drug with these properties may be beneficial both in hypertension and in heart failure. Whilst the efficacy of VPIs in hypertension has been consistently demonstrated in pre-clinical and clinical studies, the role of VPIs, if any, in heart failure is less clear, since numerous small studies have produced conflicting results. Furthermore, preliminary results from the recently completed Omapatrilat Versus Enalapril Randomised Trial of Utility in Reducing Events (OVERTURE) study have failed to establish the VPI, omapatrilat, as a first line therapy in the treatment of chronic heart failure. We review the literature on VPIs in heart failure and discuss possible reasons for the reported lack of benefit over ACE inhibitors. PMID- 12563566 TI - Stimulation of collagen gel contraction by angiotensin II and III in cardiac fibroblasts. AB - OBJECTIVE: The aim of the present study was to investigate whether angiotensin II (Ang II), angiotensin III (Ang III) or Ang II (2-8), angiotensin IV (Ang IV) or Ang II (3-8) and Ang II (1-7), Ang II (4-8), Ang II (5-8) and Ang II (1-4) can stimulate collagen gel contraction in cardiac fibroblasts in serum-free conditions. METHODS: Cardiac fibroblasts (from male adult Wistar rats) from passage 2 were cultured to confluency and added to a hydrated collagen gel in a Dulbecco's Modified Eagle's Medium, with or without foetal bovine serum, for one, two or three days. The area of the collagen gels embedded with cardiac fibroblasts was determined by a densitometric analysis. Collagen gel contraction was characterised by a decrease in the gel area. RESULTS: Ang II dose-dependently stimulated the contraction of collagen mediated by cardiac fibroblasts after one, two or three days of incubation in a serum-free medium. Telmisartan completely blocked the Ang II-induced collagen contraction by cardiac fibroblasts. PD 123319 and des-Asp(1)-Ile(8)-Ang II had no effect on the Ang II-induced collagen contraction by cardiac fibroblasts. Ang III also stimulated the contraction of collagen mediated by cardiac fibroblasts after one, two or three days of incubation in a serum-free medium. des-Asp(1)-Ile(8)-Ang II and telmisartan completely blocked the Ang III-induced collagen gel contraction by cardiac fibroblasts. des-Asp(1)-Ile(8)-Ang II, however, had no effect on the Ang II induced collagen gel contraction by cardiac fibroblasts. Ang IV and Ang II (4-8), (5-8), (1-7) and (1-4), however, had no effect on collagen gel contraction by cardiac fibroblasts. Addition of telmisartan, PD 123319 or des-Asp(1)-Ile(8)-Ang II alone did not affect collagen gel contraction by cardiac fibroblasts. CONCLUSION: Our data demonstrate that the effects of Ang II on the collagen gel contraction by adult rat cardiac fibroblasts in serum-free conditions are Ang II type 1(AT(1))-receptor- mediated, because they are abolished by the specific AT(1)-receptor antagonist, telmisartan, and not by the AT(2)-receptor antagonist PD 123319 or by the Ang III antagonist des-Asp(1)-Ile(8)-angiotensin. The Ang III stimulated contraction of collagen by cardiac fibroblasts is completely blocked by the Ang III receptor antagonist, des-Asp(1)-Ile(8)-angiotensin II, and by telmisartan. PMID- 12563567 TI - Antiproteinuric effect of candesartan cilexetil in patients with chronic glomerulonephritis. AB - A prospective, randomised, double-blind, parallel-group, dose-response trial was conducted to investigate the antiproteinuric effect of candesartan cilexetil, the angiotensin II type 1 receptor blocker, in patients with chronic glomerulonephritis. Patients (n=280) were treated for 12 weeks with candesartan cilexetil 2, 4, or 8 mg given orally once-daily (o.d.). The improvement in urinary protein excretion observed at the end of the treatment period was 15.9% in the 2 mg group, 25.6% in the 4 mg group, and 34.6% in the 8 mg group, respectively, showing a clear dose-response (2 mg <4 mg <8 mg; p=0.003). The mean reduction in urinary protein excretion was 11.3% in the 2 mg group, 26.3% in the 4 mg group, and 26.0% in the 8 mg group, showing a dose-response pattern, in that the effect of 4 mg and 8 mg was greater than that of 2 mg (2 mg <4 mg asymptotically equal to 8 mg; p=0.010). As the observed reduction in urinary protein excretion failed to correlate with changes in mean blood pressure, it could not be attributed to the antihypertensive effect of the study drug alone. This suggests that candesartan cilexetil, 4 8 mg o.d., has antiproteinuric effects in patients with chronic glomerulonephritis. PMID- 12563568 TI - Effects of telmisartan on arterial stiffness in Type 2 diabetes patients with essential hypertension. AB - Arterial wall stiffness, an important independent risk factor for cardiovascular disease in patients with hypertension, is worsened by the coexistence of diabetes mellitus. This randomised, prospective, double-blind, crossover trial assessed the effects of telmisartan on arterial stiffness in patients with Type 2 diabetes with essential hypertension. After a two-week placebo wash out period, 28 ambulatory patients received telmisartan (40 mg) or placebo for three weeks. Following a second two-week placebo wash out period, patients received the alternate treatment for a further three weeks. Augmentation index and central blood pressure (BP) were determined using the SphygmoCor device and pulse wave velocity (PWV) was measured using an automatic device, the Complior trade mark, at the beginning and the end of each period. Telmisartan significantly reduced the carotid femoral PWV compared with placebo (mean adjusted treatment difference 0.95 m/s; 95% CI: 1.67, 0.23 m/s; p=0.013). Peripheral and central diastolic, systolic and pulse pressures were also significantly reduced with telmisartan compared with placebo. In conclusion, telmisartan reduces arterial stiffness and peripheral and central BPs as assessed by PWV and pulse contour analysis in hypertensive patients with Type 2 diabetes. These properties of telmisartan suggest that it may improve cardiovascular outcome in this patient population. PMID- 12563570 TI - Mortality benefit of angiotensin-converting enzyme inhibitors after cardiac events in patients with end-stage renal disease. AB - HYPOTHESIS/INTRODUCTION: The risks and benefits of angiotensin-converting enzyme (ACE) inhibitors in patients with end-stage renal disease (ESRD) after cardiac events are unknown. We sought to determine the independent effect of ACE inhibitors (ACE-I) on long-term mortality in ESRD patients after cardiac events. MATERIALS AND METHODS: We analysed a prospective coronary care unit registry and identified 527 ESRD patients, 368 with complete data on medications prescribed, over eight years at a single, tertiary centre. RESULTS: The overall mean age was 64.4+13.8 years with 54.9% men, and 59.2% African-American. A total of 143/386 (37.0%) were prescribed ACE-I during the hospital stay for cardiac reasons, including congestive heart failure (CHF) 52.8% and acute coronary syndromes (ACS) 47.2%. There were no significant differences in the rates of hypotension or arrhythmias in those who were treated with ACE-I versus those who were not. Survival analysis over three years, adjusted for known confounders, demonstrated a 37% reduction in all-cause mortality in those who received ACE-I, (p=0.0145). CONCLUSIONS: In the setting of coronary care unit admission for CHF and ACS, ESRD patients selected for ACE-I, did not have increased rates of adverse haemodynamic or arrhythmic complications. The use of ACE-I conferred an independent mortality reduction over long-term follow-up. PMID- 12563569 TI - Involvement of the AT(2)-receptor in angiotensin II-induced facilitation of sympathetic neurotransmission. AB - Angiotensin II (Ang II) causes facilitation of sympathetic neurotransmission via prejunctionally-located AT(1)-receptors. The pithed rat is a suitable model to study the interactions between endogenously produced Ang II and the sympathetic nervous system at the peripheral level. Previously, we demonstrated that inhibition of the facilitatory actions of Ang II is a class effect of all AT(1) receptor blockers (ARB). However, all ARBs caused less than maximal inhibition after the highest dose, thus causing a U-shaped dose-response curve with respect to sympatho-inhibition. In the present study, we investigated whether the AT(2) receptor is involved in this upturn of the dose-response relationship. Accordingly, we studied the effect of the ARB, irbesartan (1 60 mg/kg), on the sequelae of electric stimulation of the thoraco-lumbar sympathetic outflow in the presence and absence of the AT(2)-blocker, PD 123319 (0.5 mg/kg +50 g/kg/min). Additionally, the effect of the combined (non- selective) AT(1)/AT(2)-receptor antagonist saralasin (0.001, 0.003, 0.01 or 0.03 mg/kg/min), on stimulation induced responses was studied. In addition, we measured PRA-levels after administration of irbesartan, in this model. The stimulation-induced increase in diastolic blood pressure (DBP) could be dose-dependently reduced by irbesartan. Co-infusion with PD 123319 increased the sympatho-inhibitory potency of irbesartan, possibly through displacement of irbesartan from plasma protein binding sites. The U-shaped dose-response relationship observed with irbesartan, which is illustrative for other ARBs in this model, was not observed when PD 123319 was co-administered with irbesartan, nor with the non-selective AT(1)/AT(2)-blocker, saralasin. PRA-levels increased from 111.0+17.8 to 198.7+22.2 ng/ml/hour after administration of irbesartan. PRA-levels did not differ when measured after the three highest doses of irbesartan. CONCLUSIONS: The present findings indicate a facilitatory role for the AT(2)-receptor, which is unmasked by the highest dose of irbesartan. Different plasma Ang II-levels are unlikely to have caused the less than maximal inhibition after the highest dose of irbesartan. PMID- 12563571 TI - Prediction of traumatic wound infection with a neural network-derived decision model. AB - The objective of this study was to develop and validate a decision model, using an artificial neural network, that predicts infection in uncomplicated, traumatic, sutured wounds. The study was a prospective, cohort study of all patients presenting to the emergency department of a county teaching hospital with uncomplicated wounds that required suturing. In evaluating and treating wounds, emergency medicine (EM) faculty and residents, resident physicians in primary-care specialties, and supervised medical students on EM clerkships followed a standardized wound-management protocol. Clinicians estimated the likelihood of subsequent infection using a 5-point scale. Wound healing was followed until sutures were removed. Wound outcome data were collected by medical personnel blinded to the initial prediction. Student's t-tests and Pearson's chi square statistic were used to identify independent predictors that served as input variables. Wound infection was the single output variable. Neural network analysis was used to assign weights to input variables and derive a decision equation. A total of 1,142 wounds were analyzed in the study. The overall infection rate was 7.2%. The most predictive factors for wound infection were wound location, wound age, depth, configuration, contamination, and patient age. To derive a decision equation for the model, the network was trained on data from half of the subjects and tested on the remainder. When used as a diagnostic test for wound infection, the decision model had a sensitivity of 70%, as compared to 54% for physicians, and a specificity of 76%, as compared to 78% for physicians. We conclude that through the use of combinations of 7 clinical variables available at the time of initial wound management, a neural network-derived decision model may be used to identify uncomplicated, traumatic wounds at higher risk for infection. PMID- 12563572 TI - Medical staff knowledge of EMTALA at a large, tertiary-care hospital. AB - The study objective was to determine what percentage of a hospital's medical staff have heard of the Emergency Medical Treatment and Active Labor Act (EMTALA) statute, and of those who have heard of it, to determine the extent of their knowledge. A questionnaire was mailed to 600 members of the active medical staff of an urban, 900-bed, private, tertiary-care hospital asking if they had ever heard of the EMTALA statute, and if so, to complete a 20-question multiple-choice quiz on specifics of EMTALA law. The main results were whether they had ever heard of EMTALA; if knowledge of EMTALA was related to specialty, age, years in practice, or frequency of ED call; and the quiz scores and any relationship they might have to those same demographic factors. Questionnaires were returned by 41.5% (n = 249). Thirty-one specialties were represented and seven had greater than 10 physicians each. Physician age and years in practice were skewed toward older ages. Approximately one-third (34.5%) took ED call at least monthly. Only 29.3% had ever heard of EMTALA. There was a significant linear correlation between on-call frequency and positive knowledge of the law, but of those who took call at least monthly, 50% had never heard of EMTALA. The average score on the 20-question quiz was 69%. Questions concerning definition of a transfer, definition of an emergency medical condition, investigations of EMTALA, on-call staff responsibilities, and the number of hospitals cited for EMTALA violations were answered correctly by <50% of the respondents. Most members of the medical staff of a large, tertiary-care hospital have never heard of EMTALA, and of those who have, the extent of their knowledge is limited. This presents challenges for the hospital to facilitate education efforts to reduce liability. PMID- 12563573 TI - Feasibility of screening and intervention for alcohol problems among young adults in the ED. AB - This study evaluates the feasibility of screening and brief intervention (SBI) for alcohol problems among young adults (18-39 years) in a rural, university ED. Research staff screened a convenience sample of patients waiting for medical treatment with the Alcohol Use Disorders Identification Test (AUDIT), used motivational interviewing techniques to counsel screen-positive patients (AUDIT >/= 6) during the ED visit, and referred patients to off-site alcohol treatment as appropriate. Patients were interviewed again at 3 months. Eighty-seven percent of age-eligible drinkers (2,067 of 2,371) consented to participate. Forty-three percent (894 of 2,067) screened positive, of which 94% were counseled. Forty percent of those counseled set a goal to decrease or stop drinking and 4% were referred for further treatment. Median times for obtaining consent, screening, and intervention were 4, 4, and 14 minutes, respectively. Project staff reported that 3% of patients screened or counseled were uncooperative. Seventy percent of 519 patients who participated in follow-up interviews agreed the ED is a good place to help patients with alcohol problems. High rates of informed consent and acceptance of counseling confirmed this protocol's acceptability to patients and indicated patients were comfortable divulging alcohol-related risk behavior. The modest times required for the process enhanced acceptability to patients as well as ED staff. The high prevalence of alcohol problems and the broad acceptance of SBI in this sample provide evidence of the ED's promise as a venue for this clinical preventive service. PMID- 12563574 TI - Predicting adverse outcomes in a diagnosis-based protocol system for rapid sequence intubation. AB - Our ED at Louisiana State University developed a unique approach to airway management by having four diagnosis-based protocols for rapid sequence intubation (RSI). This study examines protocol use and outcome from RSI in an academic ED. The study objective was to identify variables that are predictive of adverse outcomes in patients requiring RSI. This was a 4-year prospective, observational, data-gathering study of all intubations in an academic ED setting with >250,000 patient visits per year. Four protocols were established for 1) children <10 years of age, 2) adults with increased intracranial pressure, 3) adults with chronic obstructive pulmonary disease/asthma, and 4) other adults not fitting B or C. A special continuing quality improvement (CQI) committee was established to examine each case of RSI. Prospective data were collected, including age, race, gender, protocol, diagnostic group, intubation indication, and preintubation oxygen saturation. Diagnostic group was categorized as medical, blunt trauma, or penetrating trauma. Adverse outcome was defined as any case with hemodynamic changes, those requiring surgical or bronchoscopic intervention, and those requiring more than three attempts at intubation. Data were analyzed using univariate analysis, logistic regression, and a binomial regression tree analysis with SPSS 9.0 (Chicago, IL) and Answer Tree (SPSS). A total of 1,320 consecutive intubated patients were included. Protocol A was used in 4%, B in 43%, C in 15%, and D in 38%. Significant differences in number of cases with adverse outcome were based on protocol (P =.03) and final diagnosis (P <.03). Protocol C was less likely to be associated with adverse outcome than protocol D (odds ratio [OR] = 0.2, 95% confidence interval [CI] = 0.1-0.7). Penetrating trauma was more likely to be associated with adverse outcome (OR = 1.8, 95%, CI = 1.1-3.2) than blunt trauma. A regression tree analysis yielded the following, all cases using protocol A or C or medical cases using B had an adverse event in 11 of 458 (2.4%), whereas nonmedical cases using protocols B or D and medical cases using D had adverse outcomes in 73 of 862 cases (8.5%). The decision rules lead to a better classification of cases with adverse outcomes (2.4 vs 8.5%, of = 6.1%, 95% CI = 3.7-8.4). Adult trauma patients who fit the protocols B or D or adult medical patients who fit protocol B were at higher risk for adverse outcomes with RSI. This could alert the physician to a population at higher risk for adverse outcomes. Variables available in a diagnosis-based protocol RSI system can be used to predict adverse outcome among patients requiring RSI. PMID- 12563575 TI - Association between early returns and frequent ED visits at a rural academic medical center. AB - The objective of this study was to examine the influence of frequent emergency department (ED) use on early returns to the ED at a large rural academic medical center. An analysis was done of all 35,440 visits by 22,442 individuals to a large rural academic medical center ED during calendar year 2000. Of 35,440 ED visits, there were 1,992 (5.62%) return visits within 72 hours (early return). Frequent ED visits (visits made by individuals making 4 or more visits per year) was a predictor of early return visits (odds ratio [OR] 3.21, 95% confidence interval [CI] 2.93-3.52; Wald chi(2), P <.0001). Of 22,442 individuals who came to the ED during the study period, 1,601 (7.13%) returned within 72 hours. Frequency of ED use by a particular individual (4 or more visits per year) was also a predictor of early return for that individual (OR 14.55, 95% CI 12.84 16.48; Wald chi(2), P <.000001). The high rate of early returns to this rural academic ED was significantly associated with frequent visits (4 or more times per year) to the ED by particular individual. PMID- 12563576 TI - Patterns of presentation in heroin overdose resulting in pulmonary edema. AB - The study objective was to describe the morbidity of patients presenting with heroin overdose (HOD)-induced noncardiogenic pulmonary edema (NCPE) at an urban ED. A retrospective chart review of patients presenting between 1996 and 1999 with the diagnosis of HOD was conducted. Using a standardized data abstraction form, information on prehospital care, ED care, demographics, and cointoxications was collected. One hundred twenty-five charts (78%) were available for review. Of these, 13 (10%) were diagnosed with NCPE and all were male. In the field, NCPE patients had an average relative risk of 6, a Glasgow Coma Scale of 4, and all needed naloxone. The average admitted duration of use was 2.9 years for those who developed NCPE compared with 13.2 years for those who did not. Five (42%) NCPE patients tested positive for cocaine use and 7 (58%) tested positive for alcohol. In this cohort, the NCPE patients were male and less experienced users with initial low relative risk and Glasgow Coma Scale which demanded prehospital naloxone use. (Am J Emerg Med 2003;21:32-34. PMID- 12563577 TI - Supplemental oxygen is not required in trauma patients treated with IV opiates. AB - The risk of respiratory depression can prevent the proper use of opioids in trauma patients and lead to use of supplemental oxygen. However, high FiO(2) might contribute to atelectasis formation and consequently to relative hypoxia. Supplemental oxygen also can cause a risk of fire. In a randomized, controlled study we evaluated the need and effects of supplemental oxygen in 13 patients with extremity trauma who were treated pain-free with an intravenous opioid, oxycodone (dose range 6.75-13.6 mg). After opioid injection, 7 patients received 40% supplemental oxygen and 6 were breathing room air. Pulse oxygen saturation (SpO(2)), arterial blood gases, and hemodynamic parameters were monitored for 30 minutes. Atelectasis formation was evaluated with a computed tomography scan. No hypoxia, hypoventilation, or significant atelectasis formation was detected in any of the patients. Accordingly, routinely given supplemental oxygen was not considered necessary in these patients because no complications were seen. PMID- 12563578 TI - A prospective, randomized, controlled trial of benzodiazepines and nitroglycerine or nitroglycerine alone in the treatment of cocaine-associated acute coronary syndromes. AB - The purpose of the present study was to compare the use of lorazepam plus nitroglycerine (NTG) versus NTG alone in the reduction of cocaine induced chest pain in the emergency department. The secondary objective of the study was to help determine the safety of lorazepam in the treatment of cocaine- associated chest pain. The study was a prospective, randomized, single-blinded, controlled trial conducted at an university-affiliated urban emergency department (ED). All patients who presented with cocaine-associated chest pain were enrolled. Exclusion criteria included age greater than 45 years, documented coronary artery disease, chest pain of more than 72 hours duration, or pretreatment with nitroglycerin. Patients were given either sublingual nitroglycerine (SL NTG) (Group 1) or SL NTG plus 1 mg of lorazepam intravenously (Group 2) every 5 minutes for a total of 2 doses. Chest pain was recorded on an ordinal scale of 0 to 10 at baseline, and then at 5 minutes after each dose. Adverse reactions to medication were also recorded. Twenty-seven patients met the inclusion criteria and were enrolled in the study. The average age of these subjects was 34.1 years, and 67% were men. The NTG-only group consisted of 15 patients and the NTG-plus lorazepam group consisted of 12 patients. Baseline mean chest-pain scores were 6.87 in Group 1 and 6.54 in Group 2, with no differences between groups. Five minutes after initial treatment, mean scores for the two groups were 5.2 and 3.9, respectively, with a difference in means of 1.24 (95% confidence interval [CI] 0.8-3.8). Five minutes after the second treatment, the mean scores were 4.6 and 1.5, respectively, with a difference in means of 3.1 (95% CI 1.2-5). Kruskal Wallis testing showed a significant difference in pain relief between the two study groups (P =.003), with greater pain relief noted at 5 and 10 minutes in the NTG-plus-lorazepam group (P =.02 and P =.005, respectively). All patients in the study were admitted to the hospital, but no patient in either group had an acute myocardial infarction or cardiac complications in the ED. No adverse side effects were noted for either group. The early use of lorazepam with NTG was more efficacious than NTG alone, and appears to be safe in relieving cocaine associated chest pain. PMID- 12563579 TI - How distracting is distracting pain? AB - The study was to determine the effect of preexisting pain on the perception of a painful stimulus. We conducted a cross-section study at an urban ED using convenience sampling. Adult patients who had a 20-g IV catheter placed as part of their ED care were eligible for the study. Patients were excluded for the following reasons: more than one IV attempt, altered mental status, visual impairment, intoxication, or a physical abnormality at the IV site. Patients were asked to indicate on a 10-cm visual analog scale (VAS) the amount of pain they had at baseline immediately before IV placement. They were then asked to indicate on a separate VAS the amount of pain caused by the IV placement. Correlation between baseline pain and pain of the IV was assessed using Pearson's rho. One hundred patients were enrolled in the study. The pain of IV placement did not differ significantly by gender, race, who placed the IV, or the location of the IV. The correlation between baseline pain and pain of the IV placement was poor (rho =.14, confidence interval:-.06-.33). The response to a standardized painful stimulus among ED patients does not correlate highly with the severity of preexisting pain. PMID- 12563580 TI - Unique cutpoints for sitting-to-standing orthostatic vital signs. AB - The study objective was to describe the distribution of normal changes in vital signs related to moving from a sitting to a standing position. Subjects were free of cardiovascular disease and determined to be euvolemic by structured interview. Vital signs were measured after a minimum of 5 minutes of sitting and after 1 minute of standing. Among 176 subjects, mean differences in parameters (+/- standard deviation) were as follows: pulse increase, 5.3 +/- 6.6 beats per minute (bpm); systolic blood pressure (SBP) decrease, 1.2 +/- 9.8 mmHg; and shock index (SI) increase, 0.05 +/- 0.07 bpm/mmHg. These changes are less extreme than those previously published for changes from lying to standing. Unique tilt test positivity criteria and their expected specificities are as follows: pulse increase >/=20 bpm (0.98), SBP decrease >/=20 mmHg (0.97), SI increase >/=0.2 (0.99), ROSI >/=1.3 (0.95). Sitting-to-standing tilt tests require positivity criteria distinct from those published for lying-to-standing tilt tests. PMID- 12563581 TI - Patients discharged with a prescription for acetaminophen-containing narcotic analgesics do not receive appropriate written instructions. AB - To determine whether patients given prescriptions for acetaminophen-containing narcotic analgesics receive appropriate discharge instructions to reduce their intake of or stop taking other acetaminophen-containing products, we evaluated discharge instructions given by an urban, tertiary-care emergency department from September 1 to September 18, 2001 to find patients discharged while taking narcotic-analgesic compounds containing acetaminophen. We evaluated these discharge instructions to determine whether they included instructions to reduce or discontinue the use of acetaminophen compounds. Among the 1,505 discharge instructions evaluated, we found that 108 patients were discharged with a narcotic-analgesic combination product containing acetaminophen. Of these 108 patients, none of those given a prescription for such an acetaminophen-containing product was instructed to reduce or discontinue the use of other acetaminophen products. Emergency physicians, during the specified study period, did not explain to their patients the need to reduce the use of other acetaminophen containing products when prescribing acetaminophen-containing medications. Further inquiry into this potentially dangerous activity is warranted. PMID- 12563582 TI - Tetrodotoxin poisoning. AB - Tetrodotoxin (TTX) poisoning, although uncommon, is frequently seen in Taiwan, Japan, and Southeast Asia. It is rare but significant in the United States as well. Only three cases have been reported in the EM literature. We report an outbreak of six cases of TTX poisoning from eating puffer fish. On April 17, 2001, an outbreak of TTX poisoning occurred among Mainland Chinese fishermen who shared puffer fish on their boat in the Taiwan Strait. All six cases were middle aged men (aged 32-49 yr). Onset of symptoms began approximately 2 to 3 hours after ingestion; symptoms included orolingual numbness, acroparesthesia, and breathlessness. As a result of delayed transportation and initial resuscitation, one patient presented in full cardiac arrest, with recovery of spontaneous circulation after successful cardiopulmonary resuscitation. With the exception of this patient, the initial acid-base abnormalities were inconsistent with severity of illness and mild hypercapnia was common (4 out of 5). The patient who presented in full arrest died 1 day after admission as a result of intractable bradycardia (complete atrioventricular block), a finding rarely mentioned in the literature, despite intravenous atropine and dopamine infusion. The remaining patients survived without significant sequelae and were discharged after short term observation and supportive care, although some had neurologic and cardiopulmonary manifestations (muscle weakness, hypotension, hypoxemia, and hypercapnia). Some mildly hypoventilated patients recovered well without endotracheal intubation and ventilatory support. Favorable outcomes in most patients can be obtained if aggressive supportive treatment is provided in time. Thus, appropriate prehospital and ED ventilatory support (the implementation of a bag-valve mask or endotracheal intubation with good ventilatory support) is mandatory for those patients with respiratory failure. Most patients experience onset of symptoms within 6 hours of ingestion, but a few have a delayed onset up to 20 hours. Therefore, for those TTX-intoxicated patients without immediate prominent respiratory insufficiency, at least 24 hours of intensive monitoring of their respiratory state is necessary because of the different susceptibility and unpredictability of an individual course. PMID- 12563583 TI - Healthcare economics for the emergency physician. AB - Although the principles of healthcare economics are not usually part of the fundamental education of emergency physicians, an understanding of these elements will enhance our ability to contribute to improved health-care value. This article introduces the practical aspects of microeconomics, insurance, the supply and-demand relationship, competition, and costs as they affect the practice of medicine on a daily basis. Being cognizant of how these elements create a dynamic interplay in the health-care industry will allow physicians to better understand the expanded role they need to assume in the ongoing cost and quality debate. PMID- 12563584 TI - Orthopedic pitfalls in the ED: osteomyelitis. AB - Osteomyelitis can present to the emergency department as an acute, subacute, or chronic orthopedic concern. The presentation can range from subtle, as with acute hematogenous osteomyelitis in the pediatric patient population, to obvious, as with the diabetic patient who presents with a foot ulcer overlying exposed bone. Accurate early diagnosis and prompt treatment, whether with antibiotics, debridement, or both, are important in helping determine the ultimate outcome in this potential orthopedic pitfall. We review here the clinical presentation, diagnostic techniques, and management options for osteomyelitis that are applicable to the emergency practitioner. PMID- 12563585 TI - Evaluation of chest pain in the ED: factors affecting triage decisions. AB - The emergency physician's (EP) fast and correct diagnosis of patients with chest pain is crucial for preventing inappropriate discharge and dire consequences. To determine which factors affect admission decisions in the ED, we studied epidemiologic characteristics of both discharged and admitted patients, and the percentage of discharged patients who returned to the ED with acute myocardial infarction. The study included 185 patients seen in the ED because of chest pain between July 1 and 31, 1997 (every third day not included). Ninety patients were admitted: 36.7% were admitted for "observation of chest pain" and 63.3% met the criteria for active coronary heart disease. A form was used to collect personal data, medical history, risk factors, clinical examination, electrocardiogram interpretation, laboratory data, and admittance decision. EPs' diagnosis of cardiac chest pain demonstrated a sensitivity of 93.4%, a specificity of 73.4%, and a positive predictive value of 63.3%. Sensitivity for diagnosing acute myocardial infarct was 100%, with no erroneous discharges. The EP's ability to integrate the medical history information, including risk factors and pain characteristics, had a marked influence on the admittance decision. Efforts to reduce missed diagnoses are warranted. PMID- 12563586 TI - An unrecognized cause of paralysis in ED: thyrotoxic normokalemic periodic paralysis. AB - Hypokalemic paralysis associated with hyperthyroidism (TPP) is a well-known acute electrolyte and muscle function disorder. Lesser known is normokalemic periodic paralysis associated with hyperthyroidism. We describe two cases of young men with acute muscular paralysis and bilateral impairment of sensation over the lower legs who had normal plasma potassium concentrations. They were initially misdiagnosed as having Guillain-Barre syndrome or hysterical paralysis. However, thyroid function tests showed elevated serum T(3) and T(4) and markedly depressed thyroid-stimulating hormone findings consistent with hyperthyroidism. Control of the hyperthyroidism completely abolished their periodic paralysis. Thyrotoxic normokalemic periodic paralysis (TNPP) should be kept in mind as a cause of acute muscle weakness to avoid missing a treatable and curable condition. PMID- 12563587 TI - Indian salutation test in acute dorsal carpometacarpal joint dislocation of the ulnar four fingers. AB - Complete dislocation of the ulnar 4 carpometacarpal joints of the hand (excluding the thumb) is an uncommon injury. Anatomic reduction of the carpometacarpal joint (CMCJ) is essential to maintain function of the hand, and this can often be accomplished by prompt closed reduction and percutaneous Kirschner-wire fixation. The diagnosis is sometimes delayed or missed in the emergency department (ED), and a true lateral radiograph is mandatory in suspected cases. We present a case of dorsal unilateral dislocation of the ulnar 4 CMCJs without associated fracture. The injury was difficult to diagnose because of gross swelling of the hand. We describe a simple clinical test (Indian salutation test) that we found to be very helpful in raising the index of suspicion and hence increasing the likelihood of avoiding any clinical and medicolegal difficulties in such cases. A true lateral radiograph of the wrist confirmed the diagnosis, and prompt closed reduction and percutaneous Kirschner-wire fixation resulted in a good outcome in the case described here. PMID- 12563588 TI - Meeting the challenge of bioterrorism: lessons learned from West Nile virus and anthrax. AB - Hospital emergency departments (EDs) and ambulatory clinics may be the first to recognize illness related to a bioterrorist event. Every health-care institution must develop a weapons-of-mass- destruction (WMD) preparedness plan as part of its all-hazards disaster planning. As part of an all-hazards disaster plan, WMD preparedness should use the incident-command model to insure the required chain of command for effectively coordinating activities between hospital departments and external agencies. Preparedness for bioterrorism poses unique challenges. In the event of a biological attack, the hospital infection control staff and administration must already have in place the means to communicate with local and state public health agencies, the Centers for Disease Control and Prevention (CDC), local law-enforcement agencies, and the Federal Bureau of Investigation (FBI). Local and regional planners must consider how to coordinate the responses of emergency medical services (EMS), police, and fire departments with healthcare providers and the news media. Most hospitals are ill equipped to deal with a catastrophic event caused by WMD. The burden of responding to such events will fall initially on ED physicians and staff members. The severity of such an incident might be mitigated with careful planning, training and education. The responses of one hospital network to the outbreak of West Nile virus and, more recently, to the threat of anthrax, are presented as guides for bioterrorism preparedness. PMID- 12563589 TI - The intubating laryngeal-mask airway may be an ideal device for airway control in the rural trauma patient. AB - A review of the literature on advanced airway management indicates that the intubating laryngeal-mask airway (ILMA) may be an ideal device for airway control in the rural trauma patient. The ILMA is an advanced laryngeal-mask airway designed to allow oxygenation of the unconscious patient as well as blind tracheal intubation with an endotracheal tube. The ILMA is an easy-to-use airway with a high success rate of insertion, and requires little training. For the rural physician managing a difficult airway in a trauma patient, the ILMA has been found to be reliable and successful when other techniques fail, such as fiberoptic intubation and direct laryngoscopy. The ILMA has also been reported to cause less hemodynamic change and less injury to the teeth and lips than direct laryngoscopy. Further, the ILMA was found to be easier and faster to use with a higher success rate than either the combitube or endotracheal tube for unskilled healthcare providers. Limitations and complications of the ILMA may include aspiration, esophageal intubation, damage to the larynx or other tissues during blind passage of a tracheal tube, and edema of the epiglottis. PMID- 12563590 TI - Severe hypothermia caused by clonidine. PMID- 12563591 TI - Periclitoral abscess. PMID- 12563592 TI - Withdrawal syndrome from tramadol hydrochloride. PMID- 12563593 TI - Unusual presentation of acetaminophen toxicity. PMID- 12563594 TI - Dapsone use in Loxosceles reclusa envenomation: is there an indication? PMID- 12563596 TI - Overview of tropical nephrology. AB - Tropical nephrology covers renal diseases commonly seen in the tropics and elsewhere and specific tropical renal diseases seen mostly or only in the tropical area. Emphasis in this article is placed on the latter category, which includes renal involvement in tropical infectious diseases, natural toxin poisoning, and environmental renal problems. Pathologically, all renal structures can be affected. There is, therefore, a broad spectrum of pathologic changes, and clinical renal manifestations vary from mild urinary sediment changes to acute renal failure. Inflammatory processes plays an essential role in the pathogenesis of renal involvement in infection and toxin groups. Both models share the same inflammatory pathways through cytokines, chemokines, and mediators. Hemodynamic alterations, immune response, and direct nephrotoxicity are involved in the development of renal lesions. PMID- 12563597 TI - Glomerular disease in the tropics. AB - Glomerular disease in the tropics differs considerably from that in temperate climates and between the developed and the developing world. The prevalence and pattern of glomerular disease in the tropics in turn varies widely in different geographic regions and is influenced by environmental, nutritional, and socioeconomic factors. Among the infective agents that have been identified as contributing to an increased prevalence of glomerular disease in the tropics are Plasmodium malaria, Schistsomiasis mansoni, Mycobacterium leprosy, and hepatitis B virus (HBV). A decline in the prevalence can only be expected with the eradication of infections. Accurate knowledge of the prevalence, histologic features, and causes is essential to achieve prevention and management of glomerular diseases in the tropics. PMID- 12563598 TI - Malarial nephropathy. AB - Malaria is widely prevalent in the tropics. Clinically significant renal and renal-related disorders commonly occur in infection with Plasmodium falciparum and P. malariae. Falciparum malaria causes fluid and electrolyte disorders, transient and mild glomerulonephritis, and acute renal failure (ARF). It appears that ARF is mediated by a complex interaction of mechanical, immunologic, cytokine, humoral, acute phase response, nonspecific factors, and hemodynamic factors. Parasitized erythrocytes play a central role in all aforementioned pathogenic factors of ARF. Antimalarial drugs are still the cornerstone of treatment of falciparum infection. Because of the hypercatabolic state of falciparum malaria-induced ARF, hemodialysis as well as peritoneal dialysis should be immediately performed when there is a rapid increase of creatinine concentration. P. malariae, in contradistinction, can cause chronic glomerulopathy that may relentlessly progress to end-stage renal disease. Antimalarial drugs, corticosteroids, and immunosuppressive agents are not effective. PMID- 12563599 TI - Schistosomiasis and the kidney. AB - Schistosomiasis is a parasitic infection that affects 200 million people and is directly responsible for an annual death of 20,000 patients. Three species are responsible for most of the morbidity in humans: Schistosoma hematobium in Africa, S. mansoni in Africa and South America, and S. Japonicum in the Far East. Renal involvement occurs mostly with S. hematobium infection as a consequence of fibrosis and calcification of tissue-trapped ova in the lower urinary tract, leading to obstruction, reflux, infection, and stone formation. The resulting interstitial nephritis may present with tubular dysfunction syndrome before it progresses to end-stage renal disease. The bladder lesions also are precancerous. Immune complexes containing S. hematobium or S. mansoni worm antigens may deposit in the glomeruli leading to 5 classes of glomerulonephritis: mesangioproliferative; exudative; mesangiocapillary (membranoproliferative); focal segmental sclerosis; and amyloidosis. Exudative lesions occur in the presence of Salmonella coinfection. Membranoproliferative and focal segmental sclerosis correlate with the degree of associated schistosomal hepatic fibrosis, with immunoglobulin (Ig) A playing a major role in their pathogenesis. Amyloidosis occurs in prolonged infection and correlates with the antigen load. Although the acute and early chronic lesions regress under antiparasitic treatment (eg, praziquantel), chronic sequelae are irreversible. End-stage renal disease obviously requires dialysis and transplantation. PMID- 12563600 TI - Leptospiral nephropathy. AB - Renal involvement is common in leptospirosis. Clinical manifestations vary from urinary sediment changes to acute renal failure. Renal failure is observed in 44% to 67% of patients. Hypokalemia frequently occurs. Severe hypotension is an important warning sign for the later development of renal and pulmonary complications. Prognosis of the disease is generally good except for its association with pulmonary complications, especially pulmonary hemorrhage and acute respiratory distress syndrome. Interstitial nephritis is the basic renal lesion. Vasculitis is observed in the acute phase of the disease. Tubular necrosis and interstitial nephritis are responsible for renal failure. Glomerular changes usually are not remarkable. Hemodynamic alterations, immune response, and direct nephrotoxicity are responsible for the development of renal lesions. As in many infectious diseases, decreased renal blood flow and glomerular filtration rate play a basic role. Bacterial invasion and toxicity of outer membrane with generation of cytokines, chemokines, and cellular infiltration are important in cellular injury. PMID- 12563601 TI - Nephropathy associated with animal, plant, and chemical toxins in the tropics. AB - Widespread human exposure to a variety of drugs, chemicals, and biologic products and recent awareness of their toxic manifestations has led to the recognition of toxic nephropathy as an important segment of renal disease in the tropical countries. Tropical nephrotoxins are distinctly different from those seen in the rest of the world and are derived from local fauna and flora or plant and chemical sources. The spectrum of exposure varies from country to country and even from community to community, depending on variations in the distribution of local plants and animal species and prevalent social practices. Acute renal failure (ARF), either alone or in association with liver failure, neurologic abnormalities, metabolic acidosis, disseminated intravascular coagulation, or pulmonary infections is the most common form of presentation. Traditional medicines prescribed by witch doctors (traditional healers) constitute a special class of nephrotoxins among several communities in Africa and Asia. The prevalence of nephropathy caused by traditional medicines is directly related to a combination of ignorance, poverty, lack of medical facilities, lax legislation, and widespread belief in indigenous systems of medicine in rural areas. These medicines are a mix of herbs and unknown chemicals administered orally or as enemas. Clustering of cases after exposure to a particular agent suggests the possibility of a toxic insult. Common animal nephrotoxins are venoms of viper snakes, sea snakes, stinging insects, and raw gallbladder and bile of carp and sheep. Botanical nephrotoxins are encountered both in common edible plants (djenkol beans, mushrooms) and medicinal herbs (impila, cat's claw). Mistaken identification of medicinal herbs by untrained workers and even deliberate trials of toxic substitutes derived from plants frequently lead to renal disease, the most commonly reported being the Chinese herbal nephropathy. Nephrotoxicity caused by chemicals can be secondary to accidental occupational exposure in industrial work places (eg, chromic acid), or after suicidal or homicidal use (eg, copper sulphate, ethylene dibromide, ethylene glycol). Late presentation and multiorgan dysfunction are associated with a high mortality. A high index of suspicion, careful history taking, and an awareness of local practices are essential for proper diagnosis and management of toxic nephropathies in the tropics. PMID- 12563602 TI - Sickle cell disease and the kidney. AB - Sickle cell disease (SCD) affects the kidney by acute mechanisms, as a form of the sickle crisis, and insidiously with renal medullary/papillary necrosis, with resulting tubular defects. Glomerular hyperperfusion and hypertrophy results in a chronic sickle cell nephropathy that results in a significant morbidity in the progression to end-stage kidney disease. Kidney transplantation offers a major advantage to survival, and should be coupled with efforts toward prevention of recurrent disease. PMID- 12563603 TI - Renal stones in the tropics. AB - Urolithiasis is a problem that is generally increasing in the tropics as it is in most Western countries. There are 2 main types of the disorder-bladder stones in children, a form of the disorder that disappeared from Europe in the late 19th and early 20th centuries, and upper urinary tract stones in adults. The former has been decreasing in most countries in the so-called endemic bladder stone belt with gradual improvements in levels of nutrition. However, as living standards increase, particularly in the urban areas of the more affluent developing countries, so the incidence of upper urinary tract stones in adults is increasing. The types of stones formed depend mainly on the composition of urine, which, in turn, reflects the type of diet consumed in the countries concerned. The main factor that leads to the formation of bladder stones in children is a nutritionally poor diet that is low in animal protein, calcium, and phosphate, but high in cereal and is acidogenic. This leads to the formation of urine with a relatively high content of ammonium and urate ions and consequently to the formation of ammonium acid urate crystals and stones. In countries where there is also a high intake of oxalate from local leaves and vegetables, urinary oxalate is increased and, as a result, the ammonium acid urate stones often contain calcium oxalate as well. The stone problem in the tropics is compounded by low urine volumes resulting in some areas from poor drinking water, which causes chronic diarrhea, and in others from the hot climate and fluid losses through the skin. As nutrition improves in these countries, the formation of bladder stones gives way to upper urinary tract stones consisting of calcium oxalate, often mixed with calcium phosphate or uric acid, such as are formed in most Western countries. PMID- 12563604 TI - Pathology of renal diseases in the tropics. AB - Renal diseases unique to the tropics are those that occur in association with infectious diseases including dengue hemorrhagic fever, typhoid fever, shigellosis, leptospirosis, lepromatous leprosy, malaria, opisthorchiasis, and schistosomiasis. These renal complications can be classified on the basis of their clinical and pathologic characteristics into acute transient reversible glomerulonephritis, chronic progressive irreversible glomerulonephritis, amyloidosis, and acute renal failure (ARF) resulting from acute tubular necrosis, acute tubulointerstitial nephritis, and thrombotic microangiopathy. Certain primary glomerular diseases including immunoglobulin (Ig) M nephropathy and focal segmental and global glomerulosclerosis are prevalent in some tropical countries. Renal complications of venomous snakebites also are common in the tropics. This article discusses and summarizes important works in the literature in respect to the clinical syndromes, pathologic features, and pathogenesis of tropical renal diseases both in humans and experimental animal models. PMID- 12563605 TI - End-stage renal disease in the Asian-Pacific region. AB - Information on end-stage renal disease (ESRD) is important in assisting health care providers in planning renal replacement therapy. A questionnaire was sent to various countries in the Asian Pacific region and 10 countries responded. Data from Australia and New Zealand was obtained from their registry report. The questionnaire requested information on incidence, prevalence, transplantation rate, demographic data, causes of ESRD, causes of death, and mortality rates for the years 1998 to 2000. All the countries surveyed had national registries and there was a greater than 90% response rate in 7 of 12 countries. The incidence and prevalence rates of ESRD were linked to funding of dialysis, with higher reported rates in countries where dialysis was totally or heavily subsidized by the government. There was an increase in both incidence and prevalence rates between 1998 and 2000, with the mean annual percentage increase of 1.2% to 14.1% for incidence and 4.2% to 17.3% for prevalence. Diabetic nephropathy was the most common cause of ESRD in 9 of the 12 countries surveyed and 6 of the 12 countries had greater than 35% of their dialysis patients age 60 years and older. Peritoneal dialysis (PD) use varied between 3.9% to 81% of the dialysis population and reflected the health care policies of the individual countries. The transplantation rate was influenced by socioeconomic, religious, and cultural attitudes and varied between 3.1 per million population (pmp) to 32 pmp with the percentage of cadaveric transplants ranging between 0% of 85% of total transplants. Cardiovascular mortality remained the most common cause of death in the ESRD patients. Collaboration between the various national registries in the form of specific international studies may yield useful information of ESRD patients in the Asian-Pacific region. PMID- 12563606 TI - Chronic myeloid leukemia--past, present, and future. PMID- 12563607 TI - Prognosis and prognostic factors for patients with chronic myeloid leukemia: nontransplant therapy. AB - Reliable knowledge about an individual's prognosis is needed to select the appropriate treatment for patients with chronic myeloid leukemia (CML). The New CML score using age, spleen size, blast cell count, eosinophil count, basophil count, and platelet count shows good discrimination for survival (96, 65, or 42 months, P 1.20, independent of their residual renal function, were evaluated. Patients (61 women, 61 men), aged from 23 to 89 years (53.4+/-14.0 years)(+/-SD), had been undergoing MHD for one month to 17 years. INTERVENTION: Review of laboratory values and clinical outcome. MAIN OUTCOME MEASURES: Twelve-month mortality and hospitalization. RESULTS: Delivered Kt/V(sp) ranged from 1.23 to 2.71 (1.77+/ 0.34), nPNA from 0.5 to 2.15 (1.13+/-0.29 g/kg/day), and serum albumin, from 1.9 to 4.6 (3.76+/-0.37 g/dL). During the 12-month follow-up, 55 patients were hospitalized overnight at least once; 12 patients died; 5 patients underwent renal transplantation, and 6 patients left the study. The nPNA and Kt/V(sp) did not correlate significantly (r=.09) except when analysis was limited to Kt/V values < 1.5 (r=.54). Serum nPNA and albumin were the only variables with statistically significant correlations with both mortality and 3 measures of hospitalization (H): total days of H (H(D)), total number of H (H(F)), and time to first H (H(T)). The case-mix adjusted correlations for serum albumin and nPNA versus total days (r(HD)) and frequency of H (r(HF)) were significant, and Cox analysis based on H(T) and time to death resulted in significant odds ratios for each standard deviation decrement for both serum albumin and nPNA. Serum total iron binding capacity (TIBC) and creatinine concentrations also correlated with some but not all outcome measures: lower serum concentrations of these values were each significantly associated with poor clinical outcomes. CONCLUSIONS: Both nPNA and serum albumin predict prospective hospitalization and mortality in MHD patients with Kt/V > 1.20. Serum TIBC and creatinine concentrations appear to have association with some outcome measures as well. These data are consistent with the possibility that protein intake affects the clinical course even in the setting of an adequate to high hemodialysis dose. Studies based on randomized assignments to different protein intakes would be helpful to confirm these conclusions. PMID- 12563620 TI - Modifications in cyclosporine (CsA) microemulsion blood concentrations by olestra. AB - OBJECTIVE: Determine whether olestra alters the absorption of cyclosporine microemulsion in pediatric renal transplant recipients. DESIGN: Prospective, open label, crossover pharmacokinetic study. SETTING: General clinical research center in a university medical setting providing tertiary care. PARTICIPANTS: Seven pediatric-adolescent renal transplant recipients, ages 9 to 18, 5 to 24 months post-transplant with mean serum creatinine of 0.9 mg/dL (range, 0.7-1.6 mg/dL). METHODOLOGY: Patients participated in 2 study periods: 1. Patients were given their usual dose of Neoral (Novartis Pharmaceuticals Corporation, East Hanover, NJ) without olestra, 2. patients were given their usual dose of Neoral combined with 0.35 g/kg (maximum of 16 g of olestra or approximately 2 ounces of Lays WOW [Frito Lay, Plano, TX] potato chips). The 2 study periods were separated by a minimum 7-day washout period. CsA blood concentrations were obtained at 1, 2, 3, 4, 6, 8, and 12 hours after drug administration. RESULTS: Each patient in the study had a consistent decrease in area under the curve (AUC) when given olestra along with their usual dose of Neoral, compared with giving Neoral alone (5,018 ng*hr/mL versus 4,086 ng*hr/mL; P <.001). There also was a decrease in maximum concentration (Cmax) when Neoral was given with olestra compared with giving Neoral alone (1,202 ng/mL versus 876 ng/mL; P =.015). There was no statistical difference in the mean elimination rate or the trough values for both regimens (half-life 4.767 hours versus 4.771 hours and trough levels of 143 ng/mL versus 124 ng/mL). CONCLUSION: Olestra decreases total CsA exposure in pediatric renal transplant recipients. The noted decrease in AUC was not adequately predicted by CsA trough values which could lead to rejection episodes in the clinical setting. PMID- 12563621 TI - Nutritional status assessed from anthropometric measures in the HEMO study. AB - OBJECTIVE: Anthropometric methods are screening techniques for assessing nutritional stores of fat and lean tissues among persons with renal disease. This report presents cross-sectional baseline data on anthropometric indicators of nutritional status from a group of hemodialysis patients in a multicenter clinical trial, the Hemodialysis (HEMO) Study. DESIGN: The HEMO Study is a prospective, multicenter, randomized, 2 x 2 factorial clinical trial to evaluate the efficacy of the delivered dose of dialysis, defined by Kt/V, and membrane flux in reducing morbidity and mortality in (maintenance) hemodialysis patients. Standardized measures of weight, stature, body mass index (BMI), arm and calf circumference, and triceps and subscapular skinfolds were taken immediately after dialysis. The analytic methods consisted of univariate statistics, including means, standard deviations, and selected percentiles presented as tables of descriptive statistics. Study findings were compared with corresponding national reference data from the Second National Health and Nutrition Examination Survey (NHANES II). PATIENTS: Eligible patients between 18 and 80 years of age on chronic hemodialysis for at least 3 months, receiving hemodialysis 3 times per week and with a residual renal clearance of < 1.5 mL/min were examined. Patients also had to be able to attain an eKt/V of > or = 1.45 in 4.5 hours or an anthropometric volume < 45 to 50 L thus excluding persons with body weights over about 85 kg. The study sample consisted of the first 1,000 randomized patients, 464 men and 536 women; 642 blacks, 318 whites; and 40 of other racial backgrounds out of 1,847 randomized. RESULTS: Differences in nutritional status by sex, race, duration of dialysis, and comorbid disease were found among these patients enrolled in the HEMO Study. In comparison with NHANES II, these hemodialysis patients were, on average, lighter with less adipose and muscle tissue than healthy persons of the same ages. These findings can be indicators of persons with chronic disease. Those with diabetes were overweight based on their BMI values. CONCLUSION: These HEMO Study data provide a clinical reference for the use of these anthropometric indicators in assessing the nutritional status of contemporary hemodialysis patients weighing < 85 kg. PMID- 12563622 TI - Effects of an oxalate load on urinary oxalate excretion in calcium stone formers. AB - OBJECTIVE: To investigate the oxalate intake and the effect of an oxalate load on urinary oxalate excretion in calcium stone-forming (CSF) patients. DESIGN: Prospective study. SETTING: University-affiliated outpatient Renal Lithiasis Unit. PATIENTS AND CONTROLS: Seventy (70) CSF and 41 healthy subjects (HS) collected a 24-hour urine sample and were submitted to a 3-day dietary record to determine mean oxalate (Ox), calcium (Ca) and vitamin C intake. Fifty-eight (58) CSF patients were randomly selected to receive milk (N = 28) or dark (N = 30) chocolate as an oxalate load. INTERVENTION: Administration of either milk (94 mg Ox + 430 mg Ca) or dark chocolate (94 mg Ox + 26 mg Ca) for 3 days. A 24-hour urine sample was obtained before and after the load to determine calcium, oxalate, sodium, potassium, urea, and creatinine. MAIN OUTCOME MEASURE: Oxalate intake and excretion. RESULTS: CSF patients presented mean Ox intake of 98 +/- 137 mg/d, similar to that of HS (108 +/- 139 mg/d). Mean Ox and vitamin C intake was directly correlated with Ox excretion only in CSF. The consumption of dark chocolate induced a significant increase in mean urinary Ox (36 +/- 14 versus 30 +/- 10 mg/24 hr) not observed in the milk chocolate group. Thus, a 2-fold increase in Ox intake in this population of CSF patients produced a significant 20% increase in oxaluria, not observed when Ca was consumed simultaneously. CONCLUSION: The present study suggests that even small increases in Ox intake affect oxalate excretion and the mitigation of urinary oxalate increase by Ca consumption reinforces that Ca and Ox intakes for CSF patients should be in balance. Further studies are necessary to assess whether or not a 20% increase in oxaluria will lead to a higher risk of stone formation. PMID- 12563624 TI - High prevalence of overweight in a stable Spanish hemodialysis population: a cross sectional study. AB - OBJECTIVE: In the general population, the prevalence of overweight is high and is considered a mortality risk factor. In maintenance hemodialysis (MHD) patients reports regarding overweight and its predictors are scarce. Our aim was to evaluate the prevalence and predictors of overweight in MHD patients, supplemented with additional follow-up data on mortality. METHODS: Retrospective, observational, cross-sectional study of 190 white, noncomplicated patients on MHD recruited from 5 Spanish dialysis centers. Three anthropometric indexes were scored (relative body weight, skinfold thickness, and midarm muscle circumference), and body mass index (BMI) and dietary intake during a 5-day period were recorded. Patient survival was evaluated during a mean follow-up period of 25 +/- 20 months. RESULTS: Undernutrition (score < 7) was detected in only 15% of patients, and no patient had severe malnutrition (score < 4). The percentage of patients scored below 7, was similar in nondiabetics and type 2 diabetic patients, whereas it was significantly higher in type 1 diabetics (P =.002). Notably, 38% of patients (38% of nondiabetics, 50% of type 2 diabetics, and none of the type 1 diabetics) were overweight (BMI > or = 25 kg/m(2)). To evaluate the predictors of overweight, a stepwise logistic regression analysis was performed entering age, sex, time on dialysis, caloric intake normalized for ideal energy requirements, and protein intake. Overweight was independently influenced only by ageing (odds ratio [OR], 1.04; confidence interval [CI], 1.02 1.07; P =.0007) and female gender (OR, 2.05; CI, 1.09-3.86; P <.0001). By Cox proportional multivariate analysis, survival was positively influenced by BMI (RR, 0.88, CI, 0.79-0.97; P <.01). As expected, albumin also had a positive influence whereas age and diabetes had a negative influence on survival. This preliminary result suggests that a higher BMI may exert a protective role on survival. CONCLUSION: Overweight represents the predominant nutritional abnormality in our MHD population, especially in the elderly, women, and type 2 diabetics. PMID- 12563623 TI - Oral use of iron with vitamin C in hemodialyzed patients. AB - OBJECTIVE: To investigate if oral use of Sorbifer Durules (EGIS Pharmaceutical Ltd, Budapest, Hungary) (1 tablet/d) is adequate for the maintenance of serum iron and vitamin C in normal range during recombinant human erythropoietin treatment in hemodialyzed patients. One tablet of Sorbifer Durules contains 100 mg of Fe(2+) and 60 mg of vitamin C. DESIGN: Short-term, open-label clinical trial. SETTING: Hemodialysis units. PATIENTS: Twenty-four adult patients with end stage renal disease on hemodialysis. INTERVENTION: Four-week treatment period of Sorbifer Durules, preceded and followed by iron and vitamin C washout periods. MAIN OUTCOME MEASURE: Fasting predialysis serum samples were collected on days 0, 28, 56, and 84 to determine hematocrit, blood hemoglobin, serum iron, total iron binding capacity, transferrin saturation, ferritin, vitamin C, and plasma oxalate. RESULTS: Four-week treatment in hemodialyzed patients by Sorbifer Durules led to significant increase of hematocrit, blood hemoglobin, serum iron and vitamin C. This treatment did not influence the level of plasma oxalate. CONCLUSION: Oral dose of one tablet of Sorbifer Durules per day is adequate for the maintenance of serum iron in normal range during recombinant human erythropoietin treatment in hemodialyzed patients. This treatment simultaneously prevented the development of serum vitamin C deficiency and did not lead to further increase of plasma oxalate in these patients. PMID- 12563626 TI - Message from the chairperson. PMID- 12563625 TI - [Patient information. List of foods low in phosphorus and potassium. 1988]. PMID- 12563630 TI - Effectiveness of rheumatoid hand surgery: contrasting perceptions of hand surgeons and rheumatologists. AB - PURPOSE: Surgical management of rheumatoid hand diseases is controversial with large variation in practice pattern in the U.S. The purpose of this study is to evaluate the attitudes of hand surgeons and rheumatologists toward the effectiveness of rheumatoid hand surgery. METHODS: We designed a survey instrument to examine physicians' opinions about the effectiveness of different surgical treatments for rheumatoid hand deformities. The self-administered survey was mailed to a national random sample of 500 members of the American Society for Surgery of the Hand and 500 members of the American College of Rheumatology. RESULTS: Of survey responders, 82.5% of hand surgeons versus 34.1% of rheumatologists believe metacarpophalangeal joint arthroplasty improves hand function; 93.2% and 54.6%, respectively, believe prophylactic extensor tenosynovectomy prevents tendon rupture; and 52.5% and 12.6%, respectively, believe small joint synovectomy delays joint destruction. CONCLUSIONS: Rheumatologists view rheumatoid hand surgery as significantly less effective than do hand surgeons, which highlights the disagreements between the 2 specialties about the management of this clinical problem. PMID- 12563632 TI - Grade III radial collateral ligament injuries of the thumb metacarpophalangeal joint: treatment by soft tissue advancement and bony reattachment. AB - PURPOSE: The purpose of this study is to report the results of intact radial collateral ligament (RCL) soft tissue sleeve advancement with bony reattachment without additional soft tissue reinforcement as the sole surgical repair technique for the treatment of grade III RCL injuries to the thumb metacarpophalangeal (MCP) joint in acute, chronic, and late cases. METHODS: During a 20-year period, 38 of 45 patients (84%) with grade III RCL injuries were found to have tears at or adjacent to the metacarpal head (25 patients) or the proximal phalanx (13 patients) and underwent surgical repair at a mean of 10.8 months after injury. Patients were evaluated at a mean follow-up time of 3.8 years (minimum, 1 year), comparing pre- and postoperative standard thumb x-rays, varus (adduction) MCP joint stress x-rays, joint range of motion, pinch and grip strengths, clinical deformity, and subjective symptoms. RESULTS: Thirty-three patients (87%) were symptom free. Thirty-five patients (92%) regained normal pinch and grip strength. Thirty patients (79%) had full range of motion at the MCP joint, and no patient lost any interphalangeal joint motion. No recurrent symptomatic MCP joint instability was noted. Three patients (8%) had mild residual asymptomatic MCP joint volar subluxation. One patient had progression of pre-existing degenerative joint changes. CONCLUSION: Acute, chronic, and late grade III RCL instability of the thumb MCP joint can be successfully treated in the majority of cases by RCL soft tissue sleeve advancement and bony reattachment alone without the need for other soft tissue reinforcement. PMID- 12563633 TI - The relationship between basal joint arthritis and carpal tunnel syndrome: an MRI pilot study. AB - PURPOSE: This investigation explored 2 questions. First, does basal joint arthritis lead to morphologic alterations that significantly narrow the carpal tunnel? Second, does trapezial excision alter the morphology of the carpal tunnel and decompress the median nerve? METHOD: Four patients with basal joint arthritis alone were treated with ligament reconstruction and tendon interposition (LRTI) arthroplasty. Three patients with basal joint arthritis and carpal tunnel syndrome were treated with LRTI and carpal tunnel release. Preoperative and postoperative magnetic resonance imaging (MRI) scans were obtained to assess differences in carpal tunnel volume and morphology between the groups before and after surgery. RESULTS: The difference in preoperative carpal tunnel volume between groups was not significantly different. Carpal tunnel volume increased by 7% with LRTI and by 24% after LRTI and carpal tunnel release. The anteroposterior diameter of the carpal canal increased in both groups to allow a volar migration of the median nerve. CONCLUSION: LRTI increases the anteroposterior diameter and volume of the carpal tunnel. A larger comparative study would be necessary to determine whether LRTI is sufficient to decompress the carpal tunnel. PMID- 12563634 TI - Predictors for return to work in patients with median and ulnar nerve injuries. AB - PURPOSE: One of the consequences of median and ulnar nerve trauma is delayed return to work. The aim of this study was to determine return to work (RTW) and risk factors for delayed RTW in addition to time off work (TOW). Differences among median, ulnar, and combined median-ulnar nerve injuries were examined. METHOD: In this study 96 patients who were employed at the time of injury and who had undergone surgery for median, ulnar, or combined nerve injuries between 1990 and 1998 were evaluated. The response rate was 84% (n = 81). RESULTS: Within 1 year after injury, 59% (n = 48) returned to work. Mean TOW was 31.3 weeks. Return to work after combined nerve injuries was 24% versus after isolated median (80%) and ulnar (59%) nerve injuries. Level of education, type of job, and compliance to hand therapy were predictors for RTW. Furthermore, grip strength loss, tip pinch strength loss, and sensory recovery differed strongly between the RTW and no-RTW population. CONCLUSIONS: The predictors found in this study increase our understanding of delayed RTW after median and ulnar nerve injury and may be used to optimize postinjury rehabilitation. PMID- 12563635 TI - Three tendon transfer methods in reconstruction of ulnar nerve palsy. AB - PURPOSE: This study was designed to investigate the efficacy of 3 different tendon transfer techniques in restoring grip strength, correcting claw hand deformity, and improving hand function after irreparable ulnar nerve palsy. METHOD: A total of 44 patients were assessed 14 to 96 months after surgery. Twenty-four patients were reconstructed with the flexor digitorum 4-tail (FDS 4 tail) procedure, 11 with the extensor carpi radialis 4-tail (ECRL 4-tail) procedure and 9 with Zancolli's Lasso procedure (ZLP) with mean paralysis times of 47, 51, and 32 months, respectively. Grip strength measurements, improvement in active range of motion at the PIP and wrist joints, patients' ability to fully open and close their hands, as well as the sequence of phalangeal flexion were analyzed. RESULTS: Age, sex, mean follow-up duration, and surgical technique did not relate statistically to the functional outcome. Preoperative extensor lag of the proximal interphalangeal (PIP) joint and mean paralysis time, however, significantly affected the functional outcome. The ZLP and the ECRL 4-tail were found to be the most effective technique in restoring grip strength. The FDS 4 tail procedure, however, was the most successful in correcting the claw hand deformity, especially in long-standing paralysis in which there was elongation of the extensor apparatus. CONCLUSIONS: In short-term paralysis in which patients predominantly need grip strength and claw finger deformity correction, the ZLP or ECRL 4-tail procedures are recommended. In long-standing cases with extensor lag, asynchronous finger motion should be corrected with the FDS 4-tail procedure. PMID- 12563636 TI - Acute posterior interosseous nerve palsy caused by septic arthritis of the elbow: a case report. AB - A rare case of acute posterior interosseous nerve palsy caused by septic elbow arthritis is reported. The nerve was compressed beneath the arcade of Frohse by hypertrophied synovium and joint fluid at the anterior aspect of the radial neck. Decompression of the nerve, synovectomy, and irrigation of the elbow joint were done. effective. Six months after the surgery the nerve palsy had recovered completely. Two years after surgery there was no recurrence of the infection or nerve palsy. PMID- 12563637 TI - Demyelinating focal motor neuropathy of the ulnar nerve masquerading as compression in Guyon's canal: a case report. AB - Ulnar nerve-innervated intrinsic muscle weakness, in the absence of sensory complaints or deficits, usually is the result of compression at the ulnar nerve in zone II of Guyon's canal. In rare instances the problem is not caused by a compressive neuropathy but by a demyelinating focal motor neuropathy. Demyelinating neuropathies have been well documented in the neurologic literature but they have received little attention in the hand surgery literature. We report on one such case and the importance of differentiating the 2 neuropathies. Although surgery often is necessary for a compressive neuropathy it is contraindicated for a demyelinating neuropathy. PMID- 12563638 TI - Morphology and topography of intraosseous ganglion cysts in the carpus: an anatomic, histopathologic, and magnetic resonance imaging correlation study. AB - PURPOSE: To assess the morphology, topography, frequency, and etiology of intraosseous carpal ganglions. METHOD: Two hundred and eighty formalin fixed cadaveric wrists (mean age 80.3 +/- 9.7 years, range 40 to 101 years) were radiographed. Fifty specimens suspicious for cyst-like bone lesions underwent magnetic resonance imaging (T1w and proton density-fat-saturated images, 1.5T). Dissection, articular surface assessment, and histological examination were performed. Cystic lesions with adjacent destruction of the hyaline cartilage were classified as degenerative and were excluded. RESULTS: In 27 of the 50 specimens, 48 ganglion cysts (GC) were found. Prevalence of GC was 9.6%. Of 48 intraosseous carpal GC, 41 (85%) were in a peripheral location; 27/48 (56%) were located at the palmar carpus. GC had a macroscopic and microscopic relationship to the insertion of degenerated ligaments. They developed near longitudinally orientated bone surfaces, which serve as insertion for ligaments and are exposed to tension loading. CONCLUSIONS: Unlike degenerative cysts, ganglion cysts do not erode the hyaline articular cartilage and almost always have a continuity with the capsular ligaments. Intraosseous carpal ganglion cysts are probably the result of mucoid degeneration of adjacent ligaments. PMID- 12563639 TI - Arthroscopically assisted treatment of intraosseous ganglions of the lunate: a new technique. AB - PURPOSE: A minimally invasive technique of debridement and grafting the intraosseous ganglions of the lunate was investigated. MATERIALS AND METHODS: Eight patients with persistent symptoms and typical radiographic and bone scan findings were assessed independently pre- and postoperatively by using a modified Green and O'Brien wrist score. The intraosseous cyst was drilled under arthroscopic and fluoroscopic guidance via either a volar or dorsal portal, depending on the position identified on the computed tomography scan. Average follow-up time was 3.8 years (range, 1-5.6 years). RESULTS: All patients returned to employment within 4 months. Wrist scores improved 34 points, from 51 to 85 points, by 1 year after surgery with trabeculation being noted within the grafting lunate. The greatest improvements were seen in visual and analog pain scores, reducing from 68.3 to 11.2, and flexion-extension arcs, which increased from 98 degrees to 114 degrees. CONCLUSIONS: The technique of debridement of intraosseous ganglions of the lunate is safe, with minimal morbidity and recurrence of symptoms during the follow-up period. PMID- 12563640 TI - Arthroscopic assessment of the volar region of the scapholunate interosseous ligament through a volar portal. AB - PURPOSE: The volar region of the scapholunate interosseous ligament is one of the key structures that maintains scapholunate alignment. It is, however, difficult to evaluate this ligament using standard diagnostic procedures, including the arthroscopy through a dorsal portal. We have performed arthroscopic evaluation of this ligament through a volar portal. METHODS: Skin incision was made at just radial of the flexor carpi radialis tendon and the portal was opened through the tendon sheath. RESULTS: The volar approach enabled us to directly observe the extent of the tear of the volar region of scapholunate interosseous ligament. A shaver is inserted through the dorsal portal, and debridement can be performed by using the volar portal vein. We have not had any complications using volar portal. CONCLUSION: Volar approach is a feasible and safe procedure to evaluate and treat the volar region of the scapholunate interosseous ligament. PMID- 12563641 TI - The Lichtman classification for Kienbock's disease: an assessment of reliability. AB - PURPOSE: The correct identification of Lichtman stage 3A and 3B Kienbock's disease is crucial for treatment purposes. The present study evaluates the reliability of the Lichtman classification, with specific attention to differentiating stage 3A and 3B. METHODS: Four reviewers evaluated wrist radiographs from 39 patients with Kienbock's disease. Radiographs were graded according to the traditional Lichtman classification, a modification of the Lichtman classification using a radioscaphoid angle of 60 degrees to subdivide stage 3, and 8 other radiographic measures. RESULTS: The interobserver reliability of the Lichtman classification was substantial (kappa coefficient 0.63), but Stage 3A was less reliably identified (kappa 0.38). The new modification of the Lichtman classification increased both the overall (kappa 0.81) and the stage 3A (kappa 0.75) interobserver reliability. CONCLUSION: The use of the radioscaphoid angle increases the interobserver reliability of the Lichtman classification. This should allow a better understanding of the clinically important distinction between stage 3A and 3B disease. PMID- 12563642 TI - In vivo analysis of carpal kinematics and comparative review of the literature. AB - PURPOSE: Techniques have been developed very recently with which it is possible to quantify accurately in vivo 3-dimensional (3-D) carpal kinematics. The aim of this study was to evaluate the feasibility of our novel 3-D registration technique by comparing our data with data found in the literature. METHOD: The right wrists of 11 healthy volunteers were imaged by spiral computed tomography (CT) during radial-ulnar deviation and 5 of those wrists were imaged also during flexion-extension motion. With a matching technique relative translations and rotations of the carpal bones were traced. We compared our in vivo results with data presented in the literature. RESULTS: We found our in vivo data largely to concur with in vitro data presented in the literature. In vivo studies revealed only larger out-of-plane motions within the proximal carpal row than described in most in vitro studies. In vivo studies also showed larger interindividual variations. CONCLUSIONS: A single functional model of carpal kinematics could not be determined. We expect that in vivo 3-D CT studies on carpal kinematics, especially when applied to dynamic wrist motion, will have future diagnostic applications and provide information on long-term results of surgical interventions. PMID- 12563643 TI - Ulna shortening osteotomy using a compression device. AB - PURPOSE: To report the functional and radiographic outcomes of a cohort of patients treated for ulnar impaction syndrome with a single technique of ulnar shortening osteotomy. METHOD: We performed ulnar shortening osteotomy on 18 consecutive patients over a 10-year period by using an oblique osteotomy and compression plating technique with an AO compression device (Synthes, Paoli, PA). There were 11 men and 7 women in the series, with an average age of 32.7 years. All patients were graded before and after surgery with the modified wrist grading system of Chun and Palmer. RESULTS: All 18 osteotomies healed over an average of 6-8 weeks. There were significant improvements in pain, function, strength, and range of motion at an average follow-up of 3 years. Thirteen wrists were graded excellent, 3 good, and 2 fair. There were no postoperative complications, however, 8 patients ultimately required plate removal for local discomfort. CONCLUSIONS: This study showed that ulnar shortening osteotomy using an oblique osteotomy and an AO compression device is easy to execute and is associated with satisfactory outcomes. Healing time is rapid and postoperative cast immobilization is not required. PMID- 12563645 TI - Wide excision of the distal ulna: biomechanical testing of a salvage procedure. AB - PURPOSE: To study changes in the relationship of the distal radius and ulna after progressive resection of the distal ulna. METHOD: Six-degree-of-freedom motion sensors were used to determine relative motion and a displacement transducer was used to determine strain changes in the interosseous membrane (IOM). These were applied in cadaveric specimens loaded to simulate wrist motion and axial compressive forces. RESULTS: Strain in the IOM decreased with increasing resection of the distal ulna. The distance between the radius and ulna decreased with increasing resection up to 20% of ulna length. CONCLUSIONS: The decreased strain in the IOM and stabilization of the distance between the radius and ulna at 20% resection level provides biomechanical support for the satisfactory clinical results seen after wide ulnar resection. Wide excision of the distal ulna may be a viable salvage alternative to creation of a one-bone forearm. PMID- 12563644 TI - Comparison of different distal radius dorsal and volar fracture fixation plates: a biomechanical study. AB - PURPOSE: To compare the biomechanical properties of 6 dorsal and volar fracture fixation plate designs in a cadaver model. METHOD: Six different plating techniques were used on surgically simulated, unstable, extra-articular distal radius fractures in fresh-frozen cadavers. Specimens were tested to failure in axial compression with the Materials Testing System machine, and were analyzed with a motion analysis system. The 6 different fixation systems studied included an AO stainless steel Pi plate (group 1), an AO titanium Pi plate (group 2), a Forte plate (group 3), a dorsally placed Symmetry plate (group 4), a volarly placed Symmetry plate (group 5), and a volarly placed SCS/V plate (group 6). RESULTS: All dorsal plates (groups 1, 2, 3, 4) failed in apex dorsal angulation and all volar plates (groups 5, 6) failed in apex volar angulation. No group developed an average angular deformity greater than 5 degrees with a load of 100 N, which compares with the physiologic loads expected with active wrist motion. Only the volarly placed SCS/V plated specimens (group 6) resisted deformation of 5 degrees or more at loads up to 250 N, which compares with the physiologic loads expected with active finger motion, and was significantly stronger and more rigid than the other 5 plate groups. CONCLUSIONS: The SCS/V plate fixation system is the most rigid of the systems tested and may offer adequate stability for the treatment of the distal radius fracture in which the anterior and/or posterior metaphyseal cortex is comminuted severely. PMID- 12563646 TI - Anatomic parameters for planning of interosseous ligament reconstruction using computer-assisted techniques. AB - PURPOSE: Longitudinal radioulnar dissociation may result when both interosseous ligament (IOL) disruption and radial head fracture occur. Although radial head salvage or arthroplasty and temporary distal radioulnar joint pinning constitute the standard treatment for this injury, IOL reconstruction has been proposed to restore more normal forearm axis mechanics. To help provide an anatomic basis for IOL reconstruction, the purpose of this study was to characterize the geometry of the central band of the IOL and simulated IOL reconstructions. METHODS: Twenty forearms free of pathology were dissected to bone-IOL-bone and computed tomography scans were taken. Computer models of radius-IOL-ulna were created from the computed tomography CT images, and computer-aided design software was used to measure key parameters for IOL reconstruction and simulate anatomic IOL reconstructions. RESULTS: The insertion site locations of the IOL central band along the radius and ulna from the wrist were 57 +/- 3% and 34 +/- 4% of bone length, respectively. The angle at which the IOL central band inserts with the ulna was 24 +/- 4 degrees, which agrees with previously reported values. We found that the minimum graft length needed to anatomically span both cortices through tunnels was 112 +/- 14 mm. CONCLUSIONS: These data will help to provide a basis for planning and performing IOL reconstruction in cases of longitudinal radioulnar dissociation. PMID- 12563647 TI - Partial olecranon excision: the relationship between triceps insertion site and extension strength of the elbow. AB - PURPOSE: Partial excision with triceps reattachment is an accepted method for treating comminuted fractures of the olecranon. The currently recommended site for reattachment is along the articular margin of the remaining olecranon process. With computer models suggesting that this may lead to weakness of elbow extension, this study was performed to evaluate different sites for triceps reattachment with respect to extension strength of the elbow. METHOD: Ten fresh frozen human cadavers were potted and the elbow placed in varying degrees of flexion (45 degrees, 90 degrees, and 135 degrees ). Incremental loads were applied to the triceps mechanism and the force generated was recorded using an Instron materials testing machine (Instron Corp., Canton, MA). A 50% olecranon excision was then performed, simulating a comminuted olecranon fracture. The triceps tendon was attached either anteriorly along the articular margin or posteriorly along the cortical margin. All specimens were tested at both insertion sites in random order. RESULTS: A significant decrease in extension strength was observed between intact specimens and anterior attachment of the triceps tendon at all angles of elbow flexion. A decrease was only observed with posterior reattachment at 90 degrees. Posterior reattachment provided greater extensor strength than anterior attachment, with greatest differences seen at increasing positions of elbow extension. CONCLUSIONS: Although anterior reattachment of the triceps tendon after partial excision of the olecranon is currently recommended by some, this results in diminished strength of the triceps mechanism. A greater mechanical advantage with increased strength is provided by a more posterior reattachment using the technique described. These differences are greatest at increasing positions of elbow extension where triceps strength is more functionally important. PMID- 12563648 TI - Congenital flexion deformity of the middle finger and sagittal band hypoplasia. AB - PURPOSE: To report a congenital anomaly of the middle finger. METHOD: Nine patients (16 digits) are reported with congenital flexion deformity of the metacarpophalangeal (MCP) joint of the middle finger. Three patients (4 digits) had isolated deformities to the middle finger and in 6 the deformity was part of congenital ulnar drift (CUD) of the hand. Three patients had Freeman-Sheldon syndrome, 2 had nonsyndromic CUD, and 1 had arthrogryposis multiplex congenita. In CUD patients the middle finger had substantially greater flexion deformity of the MCP joint in comparison with other digits. Seven patients were treated surgically and 2 were treated nonsurgically. Five of the surgical patients had bilateral middle finger involvement. RESULTS: During surgery on 12 digits sagittal band hypoplasia of varying degrees was encountered in all patients and in all patients the extensor tendon of the middle finger was underdeveloped and often ulnarly displaced. Longitudinal imbrication of the remnants of the extensor tendon and centralizing the tendon if necessary by radial sagittal band reefing improved MCP joint flexion deformity. CONCLUSIONS: Congenital middle finger-in palm deformity in our patients was caused by sagittal band and extensor tendon hypoplasia. PMID- 12563649 TI - Abductor digiti minimi musculocutaneous island flap as an opposition transfer: a case report. AB - We described the use of an abductor digiti minimi musculocutaneous (ADM) island flap as an opposition transfer. This procedure is easy and safe, and provides more of a bulge in the thenar area than the Huber transfer. This procedure can be used in the treatment of Blauth grade 2 and 3A hypoplastic thumb. PMID- 12563650 TI - Derotational osteotomy at the shafts of the radius and ulna for congenital radioulnar synostosis. AB - PURPOSE: The purpose of this study was to describe the results of a newly developed method of correction osteotomy for congenital radioulnar synostosis. With this method the osteotomy is performed at the distal one third of the radius and proximal one third of the ulna. After K-wires are inserted intramedullarly into both bones the forearm is derotated manually to the position planned before surgery followed by cast immobilization. METHODS: Four patients with an average age of 3.9 years underwent surgery with this method and were followed-up for 45.8 months. All of their forearms were fixed before surgery at over 70 degrees of pronation. RESULTS: The average correction after surgery was 65 degrees and bone union occurred at 8 weeks after surgery without any complications. The patients' ability to perform daily activities showed a marked improvement after surgery, but there was a 20 degrees loss of correction during cast immobilization in one case. CONCLUSION: This method is a simple and safe technique to derotate the forearms of the patients with congenital radioulnar synostosis that are fixed in pronation. PMID- 12563651 TI - Biomechanical evaluation of flexor tendon function after hamate hook excision. AB - PURPOSE: Hamate hook fractures are uncommon injuries for which treatment is controversial. Excision of the hamate hook is considered to be the preferred method of treatment but the effects of hamate excision are not clearly delineated. The purpose of this study was to determine what effect, if any, excision of the hamate has on flexor tendon function. METHOD: The biomechanical effects of hamate hook excision on flexor tendon function were studied in fresh cadaveric forearm specimens with wrists fixed in 3 positions (neutral, 30 degrees extension, 30 degrees extension with 30 degrees ulnar deviation). Flexor tendon force, flexor tendon excursion, and flexor tendon shift were evaluated. RESULTS: Flexor tendon force decreased after hamate hook excision (11% in neutral, 14% in 30 degrees extension, and 15% in 30 degrees extension with 30 degrees ulnar deviation). The flexor profundus tendons had a 7- to 11-mm increase in proximal tendon excursion after hamate hook excision depending on the position of the wrist, and the flexor profundus tendons of the small finger shifted 4 to 5 mm in ulnar direction. CONCLUSIONS: The hamate hook provides some biomechanical advantage for flexor tendon function and cadaveric changes in tendon force after its excision suggest that power grip may be decreased after hamate hook excision. PMID- 12563652 TI - A corrective factor for scoring small finger function using the Littler's line and area under a Gaussian curve method. AB - PURPOSE: The Littler's line and area under a Gaussian curve method for scoring digital range of motion is an extremely convenient method for evaluating functional return after injury. This method, although being sufficiently accurate for other fingers, leads to a faulty estimation of function in the small finger. METHOD: Twenty-six normal hands in 14 volunteers were scored by using the earlier described method. The statistical significance of the resulting error in small finger function was determined by using the standard error of difference between means. Corrective methods were determined and their statistical validity was established by using the same test. RESULTS: There was a consistent error in scoring of small finger flexion by about 2.26% using the earlier-described method, which was statistically significant. The error was caused by a radial deviation of the small finger on full flexion. To eliminate the error the proximal reference point of the small finger needs to be shifted radially. The amount of this shift is determined either by using the opposite hand if normal, or by an amount equal to 0.16 times the length of the small finger. CONCLUSIONS: The use of the new reference point for calculating the functional score of the small finger has added to the accuracy of a simple and realistic scoring system for flexion injuries. This eliminates a potential source of error and makes our analysis that much more reliable. PMID- 12563653 TI - Spontaneous flexor tendon rupture in the palm: the role of a variation of tendon anatomy. AB - Spontaneous flexor tendon rupture is rare. We report 2 cases that occurred in association with a common flexor digitorum profundus (FDP) tendon to the ring and small fingers. The tendon bifurcated at the midpalmar level and ruptured at the point of bifurcation. To our knowledge this tendon variation has not been reported previously and has not been associated with spontaneous tendon rupture. Both ruptures occurred in the dominant small finger of elderly men in the absence of trauma. We believe that this unusual tendon arrangement represents a potential weak point in the musculotendinous unit. We discuss the role of this anatomic variant in the pathomechanics of tendon rupture. PMID- 12563654 TI - Vascular malformation within the flexor tendon sheath. AB - We report an unusual location of a capillary vascular malformation found entirely within the flexor tendon sheath of an 8-year-old girl. Vascular malformations have been found in many locations throughout the hand but have, to our knowledge, not been reported to involve the vinculum of the flexor tendon sheath. The vascular malformation was contained completely within the sheath. Therapeutic management involved surgical excision of the vascular malformation. PMID- 12563655 TI - The Fibonacci sequence: relationship to the human hand. AB - PURPOSE: The motion path of the digits follows the path of an equiangular spiral in which a constant angle is formed by all radial vectors along the curve. This implies that the lengths of the metacarpals, proximal, middle, and distal phalanges approximate a Fibonacci sequence in which the ratio of any 2 consecutive numbers approaches the number 1.61803 (phi). This study tested the hypothesis that the metacarpal and phalangeal bone lengths follow the Fibonacci relationship. METHODS: Standardized x-rays were taken of the hands of 100 healthy volunteers. The proximal phalanx length was subtracted from the sum of the lengths of the middle and distal phalanges and the metacarpal length was subtracted from the sum of the lengths of the middle and proximal phalanges. Confidence intervals for the quotients of the measured lengths of the adjacent bones of the hand also were used for statistical analysis. RESULTS: Only 1 of 12 bone length ratios contained the ratio phi in the 95% confidence interval, that of the small finger metacarpal and proximal phalanx. The largest variability was seen in the small finger phalangeal relationships. CONCLUSION: The application of the Fibonacci sequence to the anatomy of the human hand, although previously accepted, is a relationship that is not supported mathematically. The difference between individual bone lengths as measured at the joint line and the center of rotation of the joints may explain our finding. PMID- 12563656 TI - Sterling Bunnell, MD: the founding father. AB - Sterling Bunnell, MD, was the key person in the founding of the specialty of hand surgery in the United States. In 1944 he published Surgery of the Hand, which was to remain the authoritative hand text for almost 30 years. At about the same time he organized and was the guiding light of 9 United States Army Hand Centers spread throughout the United States. For the most part the surgeons who directed these hand centers became the founders of the American Society for Surgery of the Hand. PMID- 12563657 TI - A cost-minimisation study of alternative discharge policies after hip fracture repair. AB - It is widely assumed that health care costs can be reduced considerably by providing care in appropriate health care institutions without unnecessary technological overhead. This assumption has been tested in a prospective study. Conventional discharge after hip fracture surgery was compared with an early discharge policy in which patients were discharged to a nursing home with specialised facilities for rehabilitation. We compared costs for both strategies from a societal perspective, using comprehensive and detailed data on type of residence and all kinds of medical consumption during a 4-month follow-up period. As expected, early discharge reduced the hospital stay (with 13 days, p=0.001). More patients were discharged to a nursing home (76% versus 53%). Total medical costs during follow-up were reduced from an average of euro;15338 to euro;14281, representing relatively small and not significant savings (p=0.3). There are two explanations for this unexpected result. First, costs incurred by hip fracture patients were relatively less while in hospital. Hence, nursing home costs almost equalled hospital costs per admission day. Second, compared with the conventionally discharged group early discharged patients were subjected to more medical procedures during the first post-operative days. We conclude that: (1). early discharge shifted rather than reduced costs; (2). the details of costing have a major influence on the cost-effectiveness of alternative discharge policies. PMID- 12563658 TI - Medical student indebtedness and the propensity to enter academic medicine. AB - This paper considers the potential impact of medical school indebtedness and other variables on the propensity of US doctors to enter academic medicine. Probit models provide some evidence that indebtedness reduces the likelihood that physicians will choose academic medicine as their primary activity. Nevertheless, the magnitude of this effect is not large. As indebtedness may be endogenous, the probits are rerun using an instrumental variables approach. These estimates imply that over time indebtedness may have an important impact on the propensity of physicians to enter academic medicine. PMID- 12563659 TI - The determinants of health care expenditure: testing pooling restrictions in small samples. AB - Health care expenditure has increased substantially in all Western industrialized countries in the last decades. The necessity to contain the increase in health care expenditure has motivated the analysis of its determinants to explain differences across countries and health systems. However, recent studies have questioned the use of cross section data arguing that health systems are too different to allow for such comparisons. In this paper we investigate whether this criticism is really justified. We analyze the variations of health care expenditure in OECD countries relative to income, population aging and technological change. Our analysis is based on pooled cross section data and time series. Firstly, formulating error correction models for individual countries we demonstrate that in almost all cases the investigated variables are cointegrated. Secondly, we use a bootstrap framework for inference and examine whether the influence of explanatory variables is unique across countries. Applying recursive estimation procedures we find evidence for cross country homogeneity during the period 1961-1979. In the last two decades health care dynamics become more and more country specific thus indicating divergence of health systems and the growing importance of country-specific effects in the explanation of differences in health care expenditure. PMID- 12563660 TI - Deadweight loss of bacterial resistance due to overtreatment. AB - Widespread use of antibiotics is considered the major driving force behind the development of antibiotic resistance. The benefits of exceeding the welfare maximizing level of antibiotic use are below the costs of resistance created by this excess quantity of antibiotics used, thereby resulting in a welfare deadweight loss. This paper uses a simple economic model to examine the theoretical and empirical aspects of the welfare loss generated by resistance and analyzes its policy implications. The annual deadweight loss associated with outpatient prescriptions for amoxicillin in the United States is estimated at US dollars 225 million. PMID- 12563661 TI - How measures of perception from survey data lead to inconsistent regression results: evidence from adolescent and peer substance use. AB - In studies of peer group behavior, the direct measure of peer group behavior is often not available, and so is replaced by perceptions from survey respondents. This study shows that regression estimators are inconsistent when the correctly measured independent variable of group behavior is replaced with perceived measures from survey respondents. The inconsistency is due to three sources: projection of own behavior onto the group, rescaling the marginal effect of the group, and simple random measurement error. We discuss why each effect may cause inconsistency, derive formulas for the probability limit to quantify their effects, and illustrate with three examples of adolescent smoking and drinking. PMID- 12563662 TI - Managed care and technical efficiency. AB - By focusing exclusively on consumer benefit, previous studies of the effects of managed care have ignored important hospital efficiency gains. This study uses the HCUP sample of hospitals for 1992-1996 to estimate a stochastic frontier model of hospital technical efficiency. After controlling for hospital and market area variables, the study finds strong evidence that increased managed care insurance in a given market is associated with improved technical efficiency in the area's hospitals, especially in tertiary cases. Using Battese and Coelli's one-stage method (1995), the coefficients estimates are more efficient than for two-stage methods found in the literature. PMID- 12563663 TI - Determining the optimal combinations of mutually exclusive interventions: a response to Hutubessy and colleagues. AB - Hutubessy and colleagues have suggested a technique for decision-makers to employ in determining the optimal combination of interventions. This technique requires the adoption of Monte Carlo simulation analysis to identify the probability that a certain program will be included in an optimal mix given the uncertainty around the program's expected costs and benefits. In this response, it will be demonstrated that this methodology can lead to potential inefficiencies arising through the dependence of such probabilities on decisions relating to other programs and the failure to consider the opportunity costs of obtaining increased health benefits. A simple alternative approach is suggested which avoids these problems. PMID- 12563665 TI - Double discounting of QALYs. AB - Quality-adjusted life-years (QALYs) calculated from time tradeoff (TTO) based preferences have a time preference component. To impose a conventional discount rate on these implicitly discounted QALYs introduces some degree of double discounting. The purpose of this study was to determine the magnitude of the double discounting effect and the effectiveness of a suggested method for avoiding double discounting in a TTO-based QALY model. Our analysis used holistic and composite preference scores obtained with the TTO technique in a prior study of four hypothetical treatment paths in type 2 diabetes. Discounted composite preference scores were significantly discrepant from holistic preference scores. Adjusting TTO-based quality weights prior to external discounting reduced the discrepancy only slightly. Since time preference effects may vary with health state context, the double discounting effect needs further investigation. PMID- 12563669 TI - Retinoids and rexinoids for the 21st century: a brave new world for arthritis. PMID- 12563670 TI - Human immunodeficiency virus infection, antiphospholipid antibodies, and the antiphospholipid syndrome. PMID- 12563671 TI - Expression of lactoferrin on neutrophil granulocytes from synovial fluid and peripheral blood of patients with rheumatoid arthritis. AB - OBJECTIVE: To analyze lactoferrin expression on synovial fluid (SF) and peripheral blood neutrophils of patients with rheumatoid arthritis (RA) and to compare it with the lactoferrin expression on neutrophils from patients with osteoarthritis (OA). METHODS: Paired samples of peripheral blood and SF were obtained from 14 patients with RA and 9 patients with OA. Lactoferrin expression was evaluated on cell surfaces by cytofluorimetric analysis utilizing both polyclonal antibodies and the monoclonal anti-lactoferrin antibody AGM 2.29. Data are presented as mean fluorescence intensity. RESULTS: In patients with RA, the expression of membrane lactoferrin was significantly increased on SF neutrophils in comparison with those in peripheral blood. This increase was found using both polyclonal antibodies and AGM 2.29 (p = 0.0001, p = 0.0017, respectively). In patients with OA, the difference was not significant. In addition, lactoferrin expression on SF neutrophils of patients with RA was significantly increased compared with that found on SF neutrophils of patients with OA (polyclonal antibodies, p = 0.0015; AGM 2.29, p = 0.005). In patients with RA, no correlation was found between lactoferrin expression and disease activity. CONCLUSION: Our results provide evidence for an activation of neutrophil granulocytes at site of inflammation in RA and indicate that lactoferrin surface expression represents a reliable neutrophil activation marker. PMID- 12563672 TI - Interleukin 1 receptor antagonist anakinra improves functional status in patients with rheumatoid arthritis. AB - OBJECTIVE: This study evaluated the benefit of anakinra, a human recombinant form of interleukin 1 receptor antagonist, on the functional status of patients with active rheumatoid arthritis (RA) despite taking maximally tolerated doses of methotrexate (MTX). METHODS: Patients (n = 419) were randomized to receive, in addition to MTX (15 to 25 mg/wk), placebo or anakinra doses of 0.04, 0.1, 0.4, 1, or 2 mg/kg once daily for 24 weeks. Functional status measured on 8 scales (dressing and grooming, arising, eating, walking, hygiene, reach, grip, and activities) was evaluated at baseline and every 4 weeks with the Health Assessment Questionnaire (HAQ). A weighted sum of the scale scores is the HAQ disability index (HAQ-DI). Primary analysis of the HAQ-DI was based on an omnibus test for a positive dose-response relationship between anakinra treatment and change in HAQ-DI from baseline to week 24. RESULTS: Patients receiving anakinra experienced rapid and sizeable improvements in their HAQ-DI scores in a dose related fashion. For patients receiving either of the 2 highest doses of anakinra, improvements in the HAQ-DI occurred by week 4 that were of a magnitude considered clinically important and statistically significantly superior to placebo. CONCLUSION: In patients with persistence of RA despite MTX therapy, treatment with anakinra results in a rapid improvement in functional status as measured by the HAQ-DI. PMID- 12563673 TI - Neutrophil migration and production of reactive oxygen species during treatment with a fully human anti-tumor necrosis factor-alpha monoclonal antibody in patients with rheumatoid arthritis. AB - OBJECTIVE: To evaluate the effects of therapy with a fully human anti-tumor necrosis factor (TNF)-alpha monoclonal antibody on the production of superoxide and other reactive oxygen species (ROS) and on the migration capacity of neutrophils in patients with rheumatoid arthritis (RA). METHODS: A total of 29 patients with active RA and 25 healthy controls participated. Assessments were performed at baseline and 2 weeks after the first administration of anti-TNF alpha. The production of ROS was studied in unstimulated conditions and after stimulation of receptor dependent (serum treated zymosan, STZ) and receptor independent (phorbol mystrate acetate, PMA) pathways by luminol enhanced chemiluminescence. As well, the PMA induced burst production of superoxide was measured using the cytochrome-c reduction assay. Potential changes in neutrophil migration to joints were assessed by scintigraphy with autologous leukocytes. RESULTS: Baseline production of ROS (both spontaneously and after STZ stimulation) and superoxide and the ex vivo chemotaxis were similar in RA patients (n = 25) and controls (n = 25) and remained unchanged after administration of anti-TNF-alpha. The production of ROS after PMA stimulation was slightly higher in patients than in controls (p = 0.04) and this difference disappeared 2 weeks after the first dose of anti-TNF-alpha (p < 0.05). The scintigraphic study showed that a single dose of anti-TNF-alpha, but not placebo, markedly decreased the influx of leukocytes to inflamed joints. CONCLUSION: In patients with RA, anti-TNF-alpha therapy rapidly decreases the influx of leukocytes into inflamed joints but does not impair neutrophil chemotaxis and production of ROS. PMID- 12563674 TI - Modified Larsen scoring of digitized radiographs in rheumatoid arthritis. AB - OBJECTIVE: To facilitate storage, retrieval, and analysis of radiographic images we assessed the validity of a film digitizer and computer based system. METHODS: A total of 101 sets of radiographs of hands and feet from patients with early rheumatoid arthritis were digitized according to standards defined by the American College of Radiology. Two blinded observers applied the modified Larsen score to the digitized images and the original radiographs. RESULTS: There was good concordance between the scoring of digitized images and radiographic films; intraclass correlation coefficient was 0.89. Regression analysis of a Bland Altman plot revealed that there was no significant systematic bias. It was noted that scoring the digital images took more than twice as long as scoring the radiographic films. CONCLUSION: Our results indicate that modified Larsen scoring of adequately digitized films on standard computer displays is valid and shows good agreement with conventional techniques. Scoring digitized images appears to require more time, but this may be outweighed by the considerable advantages of computer storage and retrieval of radiographic images. PMID- 12563675 TI - The use of disease modifying antirheumatic drugs in women with rheumatoid arthritis of childbearing age: a survey of practice patterns and pregnancy outcomes. AB - OBJECTIVE: To describe the practices of rheumatologists when prescribing the disease modifying antirheumatic drugs (DMARD) methotrexate (MTX), leflunomide (LF), etanercept (ET), and infliximab (IN) to women of childbearing age with rheumatoid arthritis (RA) and the pregnancy outcomes of patients who become pregnant while taking these medications. METHODS: A questionnaire was mailed to 600 members of the American College of Rheumatology inquiring about their perception of fetal risk, their recommendations regarding the use of birth control in women of childbearing age taking DMARD, and the pregnancy outcomes of women with DMARD exposure. RESULTS: One hundred seventy-five rheumatologists (29%) returned completed surveys. Respondents were more likely to agree that pregnancy is contraindicated in women taking MTX (95%) or LF (92.7%) than for women taking ET (38.6%) or IN (46.5%). Accordingly, most required birth control for women taking MTX (95.7%) and LF (97.3%), and fewer for women taking ET (75.4%) or IN (73.4%). A total of 65 pregnancies exposed to these DMARD were reported (MTX 38, LF 10, ET 14, IN 2, MTX and ET 1). Only 3 congenital malformations, all in the MTX group, were reported among the 52 pregnancies with known outcomes. CONCLUSION: Rheumatologists agree that there is a risk of teratogenicity with MTX and LF and usually require the use of reliable methods of birth control in women taking these medications. There is no consensus about ET and IN; however, physicians still tend to discuss reliable birth control methods with their female patients. We have confirmed there is a risk of congenital malformations with in utero exposure to MTX. No malformations were reported in infants exposed to LF, ET, or IN, but the number of reported pregnancy outcomes was small. PMID- 12563676 TI - Effect of NOS2 gene deficiency on the development of autoantibody mediated arthritis and subsequent articular cartilage degeneration. AB - OBJECTIVE: To determine the effect of NOS2 gene deletion on articular cartilage degradation in autoantibody mediated arthritis (AMA). METHODS: Female C57BL/6Ai [ko] NOS2 N5 (NOS2-/-) mice (7-8 weeks old) and the counterpart C57/Bl6 Crj mice (wild-type, WT) were studied. Arthritis was induced by intraperitoneal injection of 4 mg of an arthritogenic cocktail of 4 monoclonal antibodies raised against type II collagen twice on Day 0 and Day 1 followed by intraperitoneal injection of 50 micro g of lipopolysaccharide on Day 2. Individual limbs were scored for arthritis in 4 grades; the total maximum score per mouse was 16. Femoral condyles and tibial plateaus of both knee joints were collected on Day 15 for immunohistological studies on nitrotyrosine and matrix metalloproteinase (MMP)-3 and -9. DNA fragmentation in chondrocytes was detected by the nick-end labeling (TUNEL) method. Blood was also collected on Day 15 to determine serum levels of nitrite/nitrate and interleukin 1 beta (IL-1 beta). RESULTS: Both NOS2-/- and WT mice with AMA developed clinically apparent arthritis. In WT mice, the arthritis progressed rapidly and reached the peak score 11.4 +/- 2.9 on Day 12, whereas the arthritis in NOS2-/- mice was milder and the peak score was 7.7 +/- 2.8 on Day 13 (p < 0.05). The serum nitrite/nitrate levels, histological grades of articular cartilage degradation, and numbers of apoptotic chondrocytes and nitrotyrosine positive chondrocytes were significantly lower in NOS2-/- mice with AMA than in WT mice with AMA. Conversely, significant differences were not observed in MMP-3 or -9 expression in chondrocytes, or in serum IL-1 beta levels between these 2 groups of mice. CONCLUSION: NOS2 gene deletion did not affect the inflammatory responses, but reduced the cartilage degradation. PMID- 12563677 TI - Subcutaneous administration of polymerized-type I collagen for the treatment of patients with rheumatoid arthritis. An open-label pilot trial. AB - OBJECTIVE: To determine the efficacy, tolerance and safety of subcutaneous injections of porcine type I collagen-polyvinylpyrrolidone (PVP) in patients with rheumatoid arthritis (RA). METHODS: Eleven patients with active RA on stable therapy with methotrexate (MTX) were enrolled in a 3 month prospective and longitudinal study. Patients were treated weekly with subcutaneous injections of 0.2 ml of collagen-PVP (1.7 mg of collagen) in the 8 most painful joints. The primary endpoints included the Ritchie index (RI), swollen joint count, disease activity score (DAS), erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP). The secondary endpoints included morning stiffness, pain intensity on a visual analog scale (VAS), and the Spanish-Health Assessment Questionnaire Disability Index (HAQ-DI). Improvement was determined using American College of Rheumatology (ACR) response criteria. RESULTS: Collagen-PVP was safe and well tolerated and there were no adverse events. Patients had a statistically significant improvement (p < 0.05) in basal versus 3 month's treatment in morning stiffness (Delta -32.3, -68.6%), RI (Delta -10.2, -46.4%), swollen joint count (Delta -10.7, -71.8%), VAS (Delta -39.9, -63.8%), HAQ-DI (Delta -0.5, -48.5%), DAS (Delta -1.35, -70.5%) and ACR20, 50, and 70 (80.0%; 60.0% and 20.0% respectively). We found no differences in serologic or hematologic variables. CONCLUSION: Collagen-PVP was a safe and well-tolerated drug for the short term treatment of RA. The combination of collagen-PVP plus MTX was more efficacious than MTX alone. However, double-blind placebo-controlled phase II and III clinical trials are necessary to determine whether this drug could be useful in the longterm treatment of RA. PMID- 12563678 TI - Angiopoietins, growth factors, and vascular morphology in early arthritis. AB - OBJECTIVE: To examine angiogenic growth factors in patients with early, untreated inflammatory arthritides and controls. METHODS: Synovial membrane (SM) infiltrate and Ang1, Ang2, and vascular endothelial growth factor (VEGF) mRNA and protein expression were examined using immunohistochemistry and in situ hybridization. Synovial fluid (SF) VEGF, transforming growth factor-beta (TGF-beta 1), and tumor necrosis factor-alpha (TNF-alpha) protein were measured by ELISA. Vascular morphology was assessed at arthroscopy. RESULTS: Ang2 mRNA and protein expression was observed in early psoriatic arthritis (PsA) and rheumatoid arthritis (RA) SM. Expression of Ang2 and VEGF was significantly greater in early PsA SM and correlated strongly. SF VEGF and TGF-beta 1 concentrations were also significantly higher in early PsA compared to RA. Distinct vascular morphology, with tortuous vessels in PsA, correlated with microscopic vascular scores (r = 0.54, p = 0.005) and VEGF levels (r = 0.51, p = 0.01). Ang1 mRNA and protein expression was observed, but concentrations were markedly lower than for Ang2 and VEGF. Clinical disease activity, SM infiltration, and SF TNF-alpha concentrations were similar in both groups. CONCLUSION: This is the first report of angiopoietin expression in early inflammatory arthritis. There is a close relationship between angiopoietins, VEGF, TGF-beta, and vascular morphology. There is differential angiogenesis at an early stage of inflammation, with major pathogenic and therapeutic implications. PMID- 12563679 TI - Serum levels of pregnenolone and 17-hydroxypregnenolone in patients with rheumatoid arthritis and systemic lupus erythematosus: relation to other adrenal hormones. AB - OBJECTIVE: In patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), low levels of adrenal steroids have been repeatedly demonstrated, but the site of alteration has not been exactly described because measurements of serum pregnenolone and 17-hydroxypregnenolone (17OHPreg) together with other adrenal steroids have never been performed. METHODS: We measured serum levels of adrenal hormones such as pregnenolone, 17OHPreg, dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), progesterone (P), 17-hydroxyprogesterone (17OHP), androstenedione (ASD), and cortisol in 24 healthy controls, 24 patients with RA, and 24 patients with SLE. RESULTS: Serum levels of pregnenolone were similar in RA and SLE patients as compared to healthy controls irrespective of prior prednisolone therapy. In all RA and SLE patients (including those with prior prednisolone treatment), serum levels of all measured hormones except pregnenolone were significantly lower as compared to controls. In RA patients without prior prednisolone treatment, serum levels of 17OHPreg, DHEA, cortisol, and ASD were similar to controls, and serum levels of P, 17OHP, and DHEAS were significantly lower as compared to controls. In SLE patients without prior prednisolone treatment, serum levels of 17OHPreg and cortisol were similar, and serum levels of P, 17OHP, ASD, DHEA, and DHEAS were significantly lower as compared to controls. CONCLUSION: The primary hormone of the adrenal steroid cascade, pregnenolone, is almost normal in RA and SLE irrespective of corticosteroid treatment. In patients with RA, we believe that there is a near normal P450scc reaction and a normal double step P450c17 reaction. In SLE patients, the P450scc reaction also seems normal but the second step of the P450c17 reaction seems to be inhibited. In both diseases, cortisol levels remain relatively stable at the expense of other adrenal hormones. This study revealed distinct changes of steroid pathways that are related to the disease entities. PMID- 12563680 TI - Glutathione S-transferase M null homozygosity and risk of systemic lupus erythematosus associated with sun exposure: a possible gene-environment interaction for autoimmunity. AB - OBJECTIVE: Multiple genetic factors modulate predisposition to systemic lupus erythematosus (SLE). The glutathione S-transferase (GST) genes GSTM1, GSTT1, and GSTP1 catalyze metabolic pathways for the excretion of reactive oxygen species that may be generated by cellular oxidative stress induced by ultraviolet radiation in sunlight. We hypothesized that risk of SLE associated with occupational sun exposure is modulated by GSTM1, GSTT1, and GSTP1 genotypes. METHODS: DNA samples and occupational history were collected from 243 cases and 298 controls in the Carolina Lupus Study, a population based case-control study of patients with recently diagnosed SLE. RESULTS: There was no independent association between SLE and presence of the homozygous null GSTM1 or GSTT1 genotype, the homozygous Val/Val or heterozygous Val/Ile GSTP1 genotype, or occupational sunlight exposure. The prevalence of Ro autoantibodies was significantly increased among Caucasians with the GSTM1 null genotype (OR 2.6, 95% CI 1.0, 6.8), but was somewhat weaker among African-Americans (OR 1.5, 95% CI 0.7, 3.5). In the combined analysis of occupational sunlight exposure and GSTM1 genotype, the effect of sun exposure among Caucasians varied depending on GSTM1 genotype. There was a 3-fold increased risk (OR 3.1, 95% CI 0.9, 10.8) of SLE associated with 24 or more months' occupational sun exposure among Caucasians with the GSTM1 null genotype, but sun exposure was not associated with risk among GSTM1 positive Caucasians (OR 0.6, 95% CI 0.3, 1.5). The interaction was statistically significant (p = 0.028). CONCLUSION: Our results suggest that GSTM1 homozygous null genotype may modify the effect of occupational sun exposure on the risk of SLE in caucasians. PMID- 12563681 TI - Fatigue in patients with systemic lupus erythematosus: the psychosocial aspects. AB - OBJECTIVE: Studies to prove a relationship between fatigue and immunological, inflammatory, or other disease characteristics of systemic lupus erythematosus (SLE) have shown no consistent findings. To further elucidate the basis for fatigue in SLE, we examined the affective states, personality traits, and mental health status in an unselected group of patients with SLE. METHODS: Fifty-seven Caucasian patients with SLE were examined. Fatigue was measured by the Fatigue Severity Scale. Personality traits and psychological function were evaluated by the Minnesota Multiphasic Personality Inventory-2 (MMPI-2), the affective states by Beck Depression Inventory, and mental health status by the General Health Questionnaire version 30 (GHQ-30). RESULTS: Fatigue was closely associated with high scores on subscales Depression (D-2) and Hysteria (Hy-3) on MMPI-2 (R2 = 0.31; p = 0.0002), as well as with high scores on BDI (R2 = 0.22; p = 0.0006) and GHQ (R2 = 0.33; p < 0.0001). CONCLUSION: Fatigue does not seem to be caused by any easily recognizable single or multiple factor(s) of an inflammatory or immunological state. Our results point to fatigue being a multifaceted phenomenon where several psychosocial factors are strongly related, and indicate that fatigue is part of a complex response to chronic disease. PMID- 12563682 TI - Single photon emission computed tomography dual isotope myocardial perfusion imaging in women with systemic lupus erythematosus. II. Predictive factors for perfusion abnormalities. AB - OBJECTIVE: We have reported that 40% of patients with systemic lupus erythematosus (SLE) had abnormal myocardial perfusion studies. Here we investigated risk factors for abnormal myocardial perfusion in a cohort of women with SLE without history of coronary artery disease. METHODS: Consecutive women with SLE followed at a large lupus clinic underwent single photon emission computed tomography dual isotope myocardial perfusion imaging (DIMPI) following pharmacological stress using dipyridamole. At the time of study each patient had a clinical and laboratory assessment performed by a standard protocol. We compared traditional risk factors as well as disease and therapy related factors in those with and without perfusion abnormalities. RESULTS: A total of 129 patients were studied. The mean +/- SD age was 44.8 +/- 10.9 yrs, and mean SLE Disease Activity Index was 4.2 +/- 5.1. Forty-nine (38%) patients had an abnormality of myocardial perfusion. Factors associated with an abnormal DIMPI included current hypertension (OR 2.11, p = 0.05), elevated cholesterol ever (OR 2.51, p < 0.05), and total cholesterol:high density lipoprotein-cholesterol ratio (OR 1.96 for each increase of 1.0, p < 0.008). CONCLUSION: Myocardial perfusion abnormalities are common in women with SLE without known coronary artery disease (CAD), suggesting a high burden of subclinical CAD. Several metabolic and therapy related factors appear to be associated with the process of atherogenesis in SLE. These results suggest that SLE should be considered a predisposing factor for atherosclerosis. PMID- 12563683 TI - Item weightings for the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Disease Damage Index using Rasch analysis do not lead to an important improvement. AB - OBJECTIVE: To develop Systemic Lupus International Collaborating Clinics/American College of Rheumatology Disease Damage Index (SLICC/ACR-DI) item weightings that improve the ability of the measure to predict patient mortality in systemic lupus erythematosus (SLE). METHODS: Disease damage was measured for 738 patients followed at the University of Toronto Lupus Clinic since diagnosis. Using Rasch analysis, item weightings were determined and tested for their ability to predict death in a logistic regression model. Receiver operating characteristic (ROC) curves were produced to compare the original and weighted scales' ability to discriminate patients that died during the followup period from those who remained alive. RESULTS: The average SLICC/ACR-DI score per patient was 1.66. In total, 138 of the patients died during a mean followup of 9.2 years. A Rasch analysis derived weighting scheme using weighted domain scores (SLICC/ACR-DI weighted) was the best weighted scale, with item reliability = 94%, model mean square infit = 1.01 (STD = 0.05); model mean square outfit = 0.99 (STD = 0.3), separation 4.08. The SLICC/ACR-DI-weighted was modestly better than the SLICC/ACR DI in discriminating patients who died from those who remained alive. Using standardized scores for comparability, the SLICC/ACR-DI-weighted was better in predicting patient death than the unweighted SLICC/ACR-DI [OR(death)(SLICC/ACR-DI weighted) = 1.7 vs OR(death)(SLICC-ACR-DI) = 1.4; p < 0.005]. ROC curve analysis supports that the SLICC/ACR-DI-weighted was somewhat superior to the SLICC/ACR-DI for predicting mortality. CONCLUSION: In this test set, the SLICC/ACR-DI-weighted was modestly better in predicting death than the traditional unweighted SLICC/ACR DI. However, the SLICC/ACR-DI-weighted is more difficult to apply and the weightings appear not to have provided a clinically relevant improvement of the SLICC-ACR-DI. PMID- 12563684 TI - Homocysteine plasma concentration is related to severity of lung impairment in scleroderma. AB - OBJECTIVE: To investigate the correlation between plasma concentration of total homocysteine and pulmonary involvement in patients with limited or diffuse scleroderma (systemic sclerosis, SSc). METHODS: Seventy-one patients with scleroderma were divided into 3 groups based on pulmonary involvement: Group A comprised patients without lung involvement (9 cases); Group B patients with lung involvement of mild and moderate stages (44 cases); and Group C patients with lung involvement of severe stage and endstage (18 cases). At the time of evaluation of lung involvement all patients underwent determination of plasma homocysteine concentration. Homocysteine concentration was also measured in 30 healthy controls homogeneous for sex and age. RESULTS: In patients with scleroderma the homocysteine concentration was significantly higher than in controls (11.1 and 6.9 micromol/l, respectively; p < 0.001). We found a significant association between plasma homocysteine concentration and severity of lung involvement that was not modified by correction for age, time from the diagnosis, type of scleroderma pattern, and serum creatinine and folate levels. Homocysteine concentration progressively increases in scleroderma patients with more severe pulmonary involvement. Subjects with high homocysteine concentration (i.e., > or = 75th percentile of homocysteine concentration in patients with scleroderma without lung involvement) were mostly present in the group with the greatest lung involvement. CONCLUSION: High level of homocysteinemia is associated with an increased risk of pulmonary disease in patients with scleroderma. We hypothesize that hyperhomocysteinemia may worsen injury of the endothelium, a key lesion in scleroderma disease, favoring the development of lung involvement. Our data support the hypothesis that homocysteine could be involved in the pathogenetic process of scleroderma pulmonary involvement. PMID- 12563685 TI - NOD2/CARD15 gene mutation is not associated with susceptibility to Wegener's granulomatosis. AB - OBJECTIVE: Polymorphisms and mutations in the NOD2/CARD15 gene have been reported to increase susceptibility to Crohn's disease (CD) and the rare Blau syndrome, respectively. Both conditions are characterized by granuloma formation. We assessed the influence of variants in the CARD15 gene in another disorder characterized by granuloma, Wegener's granulomatosis (WG). METHODS: Direct DNA sequencing of the CARD15 gene was performed on 25 patients with WG, and an additional 73 patients were genotyped for the 3 CD associated variants, R702W, G908R, and fs1007. RESULTS: In the WG patients, 10 previously reported single nucleotide polymorphisms (SNP) were identified. No SNP were present in the WG patients at significantly different frequencies than the control population. CONCLUSION: Our data provide no evidence to support an association between CARD15 and WG. PMID- 12563686 TI - Male sex coupled with articular manifestations cause a 4-fold increase in susceptibility to amyloidosis in patients with familial Mediterranean fever homozygous for the M694V-MEFV mutation. AB - OBJECTIVE: Familial Mediterranean fever (FMF) is an autosomal recessive disorder characterized by attacks of fever, serositis, and a predisposition to the development of amyloidosis. The wide clinical variability of the disease has been partly attributed to MEFV allelic heterogeneity and partly to the influence of additional genetic and/or environmental modifiers. Of these, male sex was found to influence disease penetrance and susceptibility to amyloidosis. We investigated the role of sex as an independent contributor to the phenotypic profile in FMF and further defined the factors affecting disease expression and severity. METHODS: A total of 124 patients with FMF who were all homozygous for the M694V mutation, including 47 patients with nephropathic amyloidosis, were identified. A detailed chart review and physical examination were undertaken to determine demographic characteristics, history, clinical manifestations, and treatment, and we calculated the disease severity score from the Tel-Hashomer key. RESULTS: A preponderance of male patients was documented (73:51; 1.4). The overall male:female ratio was significantly higher among patients with amyloidosis (32:15; 2.1) compared to patients without amyloidosis (41:36; 1.1). FMF severity scores, independently calculated for male and female patients, were equally high (9.5 +/- 3.0 and 9.7 +/- 2.8, respectively). The frequency of arthritic attacks, significantly higher in women than men (p = 0.015), remained notably higher in male FMF patients with amyloidosis compared to male FMF patients without amyloidosis (p = 0.002). Significant correlation between arthritis attacks and amyloidosis was found (R > 0.285, p < 0.001). CONCLUSION: Susceptibility to renal amyloidosis is influenced both by sex and the occurrence of joint attacks, acting as 2 MEFV independent factors (OR 2.37, 95% CI 1.06-5.26 and OR 3.27, 95% CI 1.23-8.68, respectively). PMID- 12563687 TI - Increased soluble FAS suggests delayed apoptosis in familial Mediterranean fever complicated with amyloidosis. AB - OBJECTIVE: Interactions of the FAS with FAS ligand have been proposed as a major regulatory mechanism of immune homeostasis. Soluble FAS (sFAS) acts as a competitive antagonist to FAS, thereby inhibiting FAS mediated apoptosis. sFAS concentrations have been studied in various autoimmune diseases, with controversial results. In this cross sectional study, we investigated the role of sFAS protein in attack-free patients with familial Mediterranean fever (FMF) with and without amyloidosis. METHODS: Twelve FMF patients without amyloidosis (male/female: 7/5; median age 23.5 yrs, range 17-38), 10 FMF patients with amyloidosis (male/female: 5/5; median age 41.5 yrs, range 33-51), and 14 controls (male/female: 6/8; median age 46 yrs, range 38-57) were enrolled in the study. Serum sFAS concentrations were studied by ELISA. RESULTS: Median serum sFAS concentrations were 4630 (2580-12,270), 1338 (453-3240), and 3430 (2110-5960) pg/ml in FMF patients without amyloidosis, FMF patients with amyloidosis, and controls, respectively. Intergroup differences were all statistically significant (p < 0.05). CONCLUSION: Elevated serum sFAS concentrations in attack-free FMF patients might be due to dysregulated apoptosis of polymorphonuclear leukocytes together with the ongoing subclinical inflammatory activity. On the other hand, decreased sFAS concentrations could contribute to the augmented apoptosis together with the alterations in immune response leading to the amyloidosis. PMID- 12563688 TI - Predictors of longterm outcome in ankylosing spondylitis. AB - OBJECTIVE: To determine predictors of longterm outcome in ankylosing spondylitis (AS). METHODS: Data were collected retrospectively on constitutional and environmental factors that may predict outcome in AS in 311 patients (252 men, 81%). Univariate statistics and multivariable linear regression analyses were used to identify factors correlated with disease outcome, which was defined in terms of radiological (Bath AS Radiology Index, BASRI) and functional status (Bath AS Functional Index, BASFI). RESULTS: Disease duration, sex, and iritis are independently associated with BASRI and account for 23% (p < 0.001) of variation in radiological scores (BASRI-t), a measure that includes the hip joint in the score. Radiological hip involvement is significantly associated with higher scores of spinal radiological change (BASRI-s) (p < 0.001). Cigarette smoking, radiological status, and Bath AS Disease Activity Index score (BASDAI) are independently associated with and account for 50% of variability in functional status (p < 0.001). CONCLUSION: Much of the variability in disease severity in AS remains unexplained. All but one of the factors associated with outcome in this study are inherent. This suggests that genetic factors have a greater influence than environmental factors on radiological progression and disability in AS. It may, however, be possible to improve longterm functional outcome in AS by targeting high risk individuals early in the disease course with more aggressive management strategies and encouraging smoking cessation in all patients with AS. PMID- 12563689 TI - HLA-B27 and its subtypes in 4 Taiwanese Aborigine tribes: a comparison to Han Chinese patients with ankylosing spondylitis. AB - OBJECTIVE: We surveyed B27 and its subtypes in 5 ethnic groups in Taiwan. METHODS: Blood was obtained from 281 Aborigine people of the Atayal tribe (I Lan), 141 Paiwan (Ping-Ton), 38 Rukai (Ping-Ton), and 40 Yami (Orchid Island), and also 47 B27+ healthy Han Chinese subjects and 82 B27+ Han patients with ankylosing spondylitis (AS). HLA-B27 and its subtypes were determined by standard methods. RESULTS: A much higher prevalence of B27 was found among the Atayal Aborigines (9.2%), which was significantly different from a lower prevalence in the Paiwan (2.1%; p = 0.004) and even higher than that of Han Chinese (5.59%; p = 0.04). No blood sample from the 38 Rukai Aborigines showed any B27. B2704 was the only subtype (100%) found in the 28 healthy Aborigines and 2 Aborigine patients with AS. However, in Chinese subjects, 40 of 47 (85%) B27+ healthy subjects were B2704, and 7 of 47 (15%) were B2705. In Chinese B27+ AS patients, 77 of 82 (94%) were B2704 and 5 of 82 (6%) were B2705. No other subtypes were found. Only the Aborigines without AS carrying B2704 showed a significant difference from the Chinese without AS carrying B2704 (p = 0.041). CONCLUSION: The different prevalence of B27, but similar frequency of the B2704 subtype, between Aborigines and Han Chinese suggests Aborigines are a unique population and may originate from an Asian country, possibly mainland China. PMID- 12563690 TI - Intermittent hydrostatic pressure inhibits shear stress-induced nitric oxide release in human osteoarthritic chondrocytes in vitro. AB - OBJECTIVE: To test the effects of intermittent hydrostatic pressure (IHP) on nitric oxide (NO) release induced by shear stress and matrix macromolecule gene expression in human osteoarthritic chondrocytes in vitro. METHODS: Chondrocytes isolated from cartilage samples from 9 patients with osteoarthritis were cultured and exposed to either shear stress or an NO donor. Nitrite concentration was measured using the Griess reaction. Matrix macromolecule mRNA signal levels were determined using reverse-transcriptase polymerase chain reaction and quantified by imaging analysis software. RESULTS: Exposure to shear stress upregulated NO release in a dose and time-dependent manner. Application of IHP inhibited shear stress induced NO release but did not alter NO release from chondrocytes not exposed to shear stress. Shear stress induced NO or addition of an NO donor (sodium nitroprusside) was associated with decreased mRNA signal levels for the cartilage matrix proteins, aggrecan, and type II collagen. Intermittent hydrostatic pressure blocked the inhibitory effects of sodium nitroprusside but did not alter the inhibitory effects of shear stress on cartilage macromolecule gene expression. CONCLUSION: Our data show that shear stress and IHP differentially alter chondrocyte metabolism and suggest that a balance of effects between different loading forces preserve cartilage extracellular matrix in vivo. PMID- 12563691 TI - X-ray technologists' reproducibility from automated measurements of the medial tibiofemoral joint space width in knee osteoarthritis for a multicenter, multinational clinical trial. AB - OBJECTIVE: To determine the reproducibility of x-ray technologists, 26 in North America (NA), 24 in Europe (EU), in reliably repositioning patient's osteoarthritic (OA) knees, from computerized measurements of minimum joint space width (JSW) and reproducibility in joint repositioning, during their training for the clinical trial. METHODS: Technologists from 12 NA and 12 EU clinical radiology units received identical training, at one site on each continent, in performing the fluoroscopically assisted semiflexed knee examination and in quality control criteria (QCC) for film acceptance. Subjects recruited were 129 in NA and 70 in EU, with both knees radiographed for some subjects. Each technologist radiographed 5 OA knees and repeated the process on the same knees 2 to 7 days later. Minimum medial JSW was measured at a single center on digitized images with computer software that corrected for radiographic magnification. Technologists' reproducibility in joint repositioning and JSW measurement was determined from the difference between test and retest. RESULTS: In all, only 3/50 technologists failed qualification criteria with a repeat-film JSW difference > 0.50 mm. The mean, standard deviation (SD) of the difference in JSW between test/retest for 146 NA film-pairs of -0.020 (0.16) mm was not statistically different from that in 120 EU film-pairs: -0.001 (0.18) mm. In NA and EU 45% of examinations achieved high quality, i.e., JSW difference between repeat films < 0.1 mm, and 92% achieved excellent to good quality with a difference between repeat films < 0.3 mm. NA and EU technologists' reproducibility was unaffected by subject's sex, age, and degree of JSW loss. Reproducibility in joint reposition for all technologists was excellent. CONCLUSION: Between-continent precision of JSW measurements from all accepted pairs of semiflexed views was excellent to very good and similar to the high technical quality achieved in the authors' original report. The value of training incorporating both test/retest radiographs and film QCC is essential for the high technical quality required for multinational clinical trials. PMID- 12563692 TI - Blood-induced joint damage: longterm effects in vitro and in vivo. AB - OBJECTIVE: We previously showed that 4-day in vitro exposure of human cartilage to blood, as well as a single experimental joint bleeding in dogs, resulted in a disturbed cartilage matrix turnover lasting at least 2 weeks. We now evaluate the longterm outcome of the adverse in vitro and in vivo effects of blood on cartilage matrix turnover. METHODS: Human and canine articular cartilage tissue was cultured in the presence of homologous whole blood during 4 days. The in vitro cartilage matrix turnover was analyzed directly after blood exposure or following culture for additional periods of 2, 5, and 10 weeks in the absence of blood. The in vivo longterm effects were determined by injecting autologous blood into the right knee of 12 Beagle dogs. Six dogs were killed shortly after blood injections; the 6 remaining dogs were killed 10 weeks later. Cartilage matrix turnover and the cartilage destructive properties of the synovial tissue were analyzed. RESULTS: Short term (4 days) in vitro exposure of human or canine cartilage to whole blood inhibited proteoglycan synthesis by more than 98% (day 4), an inhibition which lasted until week 10 (70 and 75% inhibition, respectively). Also the in vivo short term exposure of cartilage to blood induced the adverse changes in cartilage proteoglycan turnover seen shortly after exposure. However, in vivo 10 weeks after the last injection, normalization of cartilage matrix turnover was observed. Synovial inflammation was absent and no destructive activity was found. CONCLUSION: These data show a discrepancy between the in vitro and in vivo longterm effects of blood on cartilage. A possible explanation for the in vivo recovery after experimental joint bleeding in dogs could be that the observed changes in cartilage only predispose to acute damage but that additional (e.g., mechanical) factors are needed to induce permanent joint damage. PMID- 12563693 TI - Enhanced lipid peroxidation in synoviocytes from patients with osteoarthritis. AB - OBJECTIVE: To evaluate the degree of lipid peroxidation of synoviocytes from patients with rheumatoid arthritis (RA), osteoarthitis (OA), and controls and to look at the production of nitric oxide (NO) and its involvement in this process. METHODS: Human synoviocytes were isolated from synovial tissues from patients with RA, OA, and from healthy controls. Cells were maintained in culture for up to 3 culture passages. Lipid peroxidation, verified by the production of malonaldehyde (MDA) and 4-hydroxy-2(E)-nonenal (4-HNE), was determined by colorimetric assay. NO was evaluated by estimating the stable NO metabolite nitrite by the Griess method in the supernatants of unstimulated and interleukin (IL)-1beta and tumor necrosis factor (TNF)-a stimulated cells. RESULTS: Increased levels of lipid peroxidation were observed for OA-derived synoviocytes compared to RA and controls. The cells in each experimental group produced low amounts of NO both in basal and in stimulated conditions. CONCLUSION: In OA, synovial cells underwent a lipid peroxidation process that did not occur in synoviocytes from RA or controls even in the absence of a detectable production of the reactive nitrogen intermediate NO. We can postulate that this peroxidation process might be due to the action of NO secreted by chondrocytes that are known to produce higher levels of this radical in OA compared to RA. PMID- 12563694 TI - Abnormalities in the replicative capacity of osteoblastic cells in the proximal femur of patients with osteonecrosis of the femoral head. AB - OBJECTIVE: Aseptic nontraumatic osteonecrosis (ON) of the femoral head is a painful disorder that often leads to femoral head collapse due to subchondral fracture. We postulated that alteration of osteoblast function might play a role in the pathophysiology of ON. We evaluated the ex vivo proliferation rate and differentiation capacity of osteoblasts derived from the intertrochanteric region of the femur and of the iliac crest of patients with ON of the femoral head and compared it with patients with hip osteoarthritis (OA). METHODS: We examined the function of osteoblastic cells in cultures derived from bone biopsies of the intertrochanteric region of the femur and of the iliac crest obtained from 13 patients with ON of the femoral head and 8 patients with hip OA. The replicative capacity was assessed by the proliferation rate in secondary culture. The phenotypic characterization was evaluated by the level of alkaline phosphatase activity, the sensitivity to 1,25 (OH)2 vitamin D3, and collagen synthesis. RESULTS: The replicative capacity of the osteoblastic cells of the intertrochanteric area of the femur in ON patients was significantly reduced compared to patients with OA. The capacity of differentiation, however, was not different between ON and OA patients. CONCLUSION: The replicative capacity of osteoblastic cells is significantly reduced in the femur of patients with ON. Our results confirm that altered osteoblastic function plays a role in the pathophysiology of ON of the femoral head. PMID- 12563695 TI - Ultrasonographic evaluation of pes anserinus tendino-bursitis in patients with type 2 diabetes mellitus. AB - OBJECTIVE: To assess musculoskeletal ultrasonographic (US) findings in patients with type 2 diabetes mellitus (DM) with and without pes anserinus (PA) tendinitis or bursitis syndrome; and to determine possible etiologic factors such as systemic diabetic microvascular disease complications in these patients. METHODS: The knee joints were examined with an ultrasound real-time scanner using a 10 MHz electronic linear transducer in 48 patients with type 2 DM and 25 controls. The presence of systemic diabetic microvascular disease complications was evaluated. RESULTS: On examination 23 (23.9%) knees of the 14 (29.1%) patients with type 2 DM were found to have PA tendinitis or bursitis syndrome. US revealed that only 4 (8.3%) of the diabetic patients with PA tendinitis or bursitis syndrome had PA tendonitis findings. There were no significant differences in the thickness of PA tendons between the diabetic patients with bilateral knee PA tendinitis or bursitis syndrome (9 patients) and controls, or between the asymptomatic and symptomatic knees in patients with unilateral PA tendinitis or bursitis syndrome (5 patients). The prevalence of morphologic changes of the medial meniscus, effusion and synovitis in the suprapatellar recess, popliteal cyst, and radiographic osteoarthritis (OA) in the diabetic patients with PA tendinitis or bursitis syndrome was found to be increased. CONCLUSION: The prevalence of PA tendinitis or bursitis syndrome is not uncommon on examination in patients with type 2 DM. However, patients with clinically diagnosed PA tendinitis or bursitis syndrome less frequently have morphologic US changes of the PA tendons. Our results also suggest that structural changes such as meniscus lesions that occur in consequence of OA might have a role in the etiology of medial knee pain in diabetic patients. PMID- 12563696 TI - Inhibition of disease progression by a novel retinoid antagonist in animal models of arthritis. AB - OBJECTIVE: To investigate the usefulness of a novel retinoic acid receptor (RAR) antagonist (BMS-189453) in animal models of arthritis. METHODS: BMS-189453 was tested in HIG-82 rabbit synovial fibroblasts to determine its ability to repress collagenase (matrix metalloproteinase-1, MMP-1) mRNA expression in vitro. Cells were stimulated with phorbol myristate acetate or interleukin 1 beta and mRNA quantified by slot-blot analysis. In vivo, BMS-189453 was evaluated in 2 animal models of arthritis: collagen induced arthritis (CIA) in mice and streptococcal cell wall induced arthritis (SCWA) in rats. Clinical scores for arthritis were recorded weekly. At the end of each study, limbs were evaluated histologically. In CIA, these results were correlated with mRNA levels for collagenase-3 (MMP-13) and stromelysin-1 (MMP-3) as determined by Northern blot. RESULTS: BMS-189453 reduced MMP-1 expression in HIG-82 synovial fibroblasts in culture. BMS-189453 treatment blocked the clinical progression of arthritis beyond soft tissue inflammation in the CIA model. In the SCWA model, BMS-189453 treatment resulted in significantly reduced swelling with no notable progression to joint distortion/destruction. Histological evaluation of the joints from animals in both models confirmed this result. Analysis of mRNA from the CIA paws showed that BMS-189453 prevented the overexpression of MMP-13 and MMP-3 in arthritic joints. CONCLUSION: Improvement in clinical and histologic variables in 2 separate animal models, along with simultaneous reduction in MMP expression in the affected joint, suggests that RAR antagonists such as BMS-189453 may be useful as agents to treat rheumatoid arthritis and for determining the role of MMP in disease progression. This is the first study to show the clinical potential of RAR antagonists in arthritis. PMID- 12563697 TI - Function of beta-adrenergic receptors on mononuclear cells in female patients with fibromyalgia. AB - OBJECTIVE: To investigate the beta-adrenergic receptors (beta AR) in patients with chronic fibromyalgia syndrome (FM). These receptors are present on circulating mononuclear cells, and activation of G-protein coupled receptors like beta AR leads to an increase in the intracellular level of cyclic aminosine monophosphate (cAMP). Therefore, cAMP levels can be used to indirectly assess the functional status of the receptor. METHODS: Eight female patients with FM and 9 matched healthy female controls participated in this study. Blood samples were drawn from subjects' anterocubital vein in the morning. Mononuclear cells were isolated from the whole blood according to Boyum's method. Basal and stimulated intracellular cAMP levels were determined by enzyme immunoassay. Aliquots of 10(6) cells were incubated with or without stimulation of beta-agonist isoproterenol for 5 min. Two different concentrations of isoproterenol (10(-3) M and 10(-5) M) were utilized. RESULTS: The basal cAMP levels in patients with FM (3.02 +/- 0.44 pmol/10(6) cells) were slightly more elevated (but not statistically different; p = 0.124, Mann-Whitney U test) than that of the control group (2.26 +/- 0.39 pmol/10(6) cells). Proterenol 10(-3) M stimulation significantly increased intracellular cAMP from the basal levels in both groups (FM group, p = 0.008; control group, p = 0.011). However, isoproterenol 10(-5) M did not increase mean intracellular cAMP levels in the FM group (p = 0.74), while a significant increase was observed in the control group (p = 0.012). CONCLUSION: These preliminary results suggest that diminished beta AR function is associated with the chronic FM state. PMID- 12563698 TI - Pain extent and diagnosis: development and validation of the regional pain scale in 12,799 patients with rheumatic disease. AB - OBJECTIVE: To develop and validate a pain scale that measures the extent of body pain. METHODS: A total of 12,799 patients with rheumatoid arthritis (RA), osteoarthritis (OA), and fibromyalgia (FM) completed a mailed survey regarding the location and intensity of their pain in 38 articular and nonarticular regions. The data were analyzed using item response theory (IRT) by nonparametric Mokken analysis followed by Rasch analysis. The resultant scale was examined for its association with clinical severity variables and its ability to distinguish patients diagnosed with and without FM. RESULTS: The resultant 19 item regional pain scale (RPS) was composed primarily of nonarticular regions. The scale had strong scalability as measured by the Mokken H statistic (H = 0.52), and satisfied the Mokken monotonicity and double monotonicity criteria. The RPS also fit the Rasch model and had satisfactory reliability and separation statistics. Of all clinical variables assessed by survey, the RPS best identified patients diagnosed with FM. In addition, the scale correlated with measures of clinical severity, regardless of diagnosis, and predicted measures of utilization. CONCLUSION: The RPS is a valid scale of pain extent. It can be useful to identify patients with FM or can be used to develop a new definition of FM, even among patients with concomitant illnesses such as RA and OA. In addition, it is a measure of pain extent that is disease independent, and works as well in RA and OA as in FM to identify patients with increased severity and resource utilization. PMID- 12563699 TI - Translation and cultural adaptation of quality of life questionnaires: an evaluation of methodology. AB - OBJECTIVE: To evaluate the methodological steps currently proposed in translation and cultural adaptation of quality of life questionnaires. METHODS: Fifty patients with rheumatoid arthritis were invited to participate. Two versions each of the Stanford Health Assessment Questionnaire (HAQ), the MOS 36 Item Short-Form Health Survey (SF-36), and the Arthritis Impact Measurement Scales 2 (AIMS-2) were administered: version 1 was a literal translation of the questionnaire; version 2 resulted from a process of translation and cultural adaptation following internationally accepted guidelines. For each patient we applied 2 questionnaires before and after consultation. The questionnaire, the order of administration, and the version were randomly assigned. The interviews were performed by a single interviewer. Several clinical and laboratory outcome measures were assessed simultaneously. Descriptive statistical analysis was used to characterize the data. Spearman and intraclass correlation coefficients were used to evaluate reliability and validity of each version of each questionnaire. RESULTS: Patients' mean (SD) age was 47 (12) years and mean (SD) disease duration was 10 (7) years. The differences of the mean in the physical and affective AIMS 2 components between the literal and adapted versions (0.21 and 0.11, respectively) were similar to the differences in the intraobserver application of the same version of culturally adapted AIMS-2 components (0.03 and 0.20) (the component scores range from 0 to 10). The same results were observed when considering other components of AIMS and SF-36, as well as HAQ scores. Version 1 and 2 presented a similar clinically and statistically significant correlation with clinical and laboratory measures used in the validation process of the questionnaires. CONCLUSION: The complex methodologies proposed in the translation and validation of the questionnaires should be carefully reevaluated. The simplification of this methodology should be studied. PMID- 12563700 TI - Prognostic factors in juvenile rheumatoid arthritis: a case-control study revealing early predictors and outcome after 14.9 years. AB - OBJECTIVE: To describe the physical and psychosocial outcome in patients with juvenile rheumatoid arthritis (JRA), compared with subjects in the general population, and to determine patient characteristics, HLA alleles, and disease variables within the first 6 months of disease onset that predict persistent disease, joint erosions, and physical disability. METHODS: A cohort of 268 (85%) of 316 patients with JRA first admitted to the hospital between 1980 and 1985 were examined after a median of 14.9 years (range 11.7-25.1) of disease duration. Controls matched for age, sex, and geographic region were randomly selected from the general population. Patients' medical records were retrospectively reviewed. Clinical examinations and radiographs of the hips, ankles, and affected joints were obtained. HLA-DRB1 and DPB1 alleles were determined by genotyping and HLA B27 by serologic testing. Physical and psychosocial health status was assessed using the Short-Form Health Survey (SF-36) and the Health Assessment Questionnaire (HAQ). RESULTS: At followup, 133 patients with JRA (50%) were in remission, 63 (24%) had developed joint erosions, and 93 (36%) had impaired physical functioning (HAQ > 0.0). Patients had greater disability, more bodily pain, and poorer general health than controls. Comparable levels of education, social function, and mental health were found, but the patients had higher rates of unemployment than controls (19% vs 7%; p < 0.001). Predictors of persistent disease and joint erosions were: young onset age and large numbers of affected joints, long duration of elevated erythrocyte sedimentation rate (ESR), and positive IgM rheumatoid factor (RF) within the first 6 months. Additionally, persistent disease was predicted by the presence of DRB1*08, and joint erosions were predicted by symmetric arthritis and DRB1*08 and HLA-B27 in combination. DRB1*01 was a predictor of joint erosions in the pauciarticular onset type (n = 163). Predictors of physical disability were: female sex, symmetric arthritis, hip joint involvement, long duration of elevated ESR and IgM RF. CONCLUSION: Compared with healthy controls, patients with JRA had impaired physical health and lower employment rates after more than 11 years of disease duration. Elevated ESR, extensive and symmetric arthritis, positive IgM RF, DRB1*08, DRB1*01, HLA B27 and DRB1*08 in combination, early onset, and female sex were early risk factors for an unfavorable outcome. PMID- 12563701 TI - Health status of patients with juvenile rheumatoid arthritis at 1 and 5 years after diagnosis. AB - OBJECTIVE: To describe the health and functional status of children with juvenile rheumatoid arthritis (JRA) diagnosed in the early 1990s. METHODS: Patients were obtained from the Pediatric Rheumatology Disease Registry, a database of patients seen in pediatric rheumatology centers across the United States. Questionnaires designed to be filled out after retrospective chart review were sent to pediatric rheumatologists caring for children diagnosed with JRA between 1992 and 1997. RESULTS: We studied 703 patients -- 376 with pauciarticular onset (pauci), 232 with polyarticular onset (poly), and 95 with systemic onset JRA (systemic). At 1 year after diagnosis, half of the pauci and systemic patients no longer required medication, compared to 78% of the poly patients; 98% of the patients functioned in Steinbrocker classes I and II. Six percent of pauci, 27% of poly, and 11% of systemic patients had limitations in school function. Nearly 1/3 of poly patients already had joint space narrowing on radiograph. By 5 years after diagnosis, all pauci, 88% of poly, and 70% of systemic patients were in Steinbrocker classes I and II; but 6% of pauci, 28% of poly, and 44% of systemic patients had limitations in school function. Nearly 2/3 of poly and systemic patients had joint space narrowing. CONCLUSION: In these children treated prior to the era of biologic therapy, at 5 years after onset, > 25% of poly and nearly half of systemic patients had functional limitations that required modifications in their school schedule. Radiographically evident joint space damage was seen within a year of onset in poly patients, and by 5 years 2/3 of poly and systemic patients had damage. PMID- 12563702 TI - Macrophage activation syndrome following initiation of etanercept in a child with systemic onset juvenile rheumatoid arthritis. AB - Systemic onset juvenile rheumatoid arthritis (JRA) accounts for 10-15% of all JRA. Macrophage activation syndrome (MAS), which can also be considered as secondary hemophagocytic lymphohistiocytosis, is a potentially life-threatening complication of systemic onset JRA. We describe a child with systemic onset JRA who developed MAS after initiation of etanercept therapy. PMID- 12563703 TI - Salazosulfapyridine-induced remission of Felty's syndrome along with significant reduction in neutrophil-bound immunoglobulin G. AB - Felty's syndrome is characterized by neutropenia, splenomegaly, and leg ulcers in patients with rheumatoid arthritis. The pathogenesis of the neutropenia is an immune-mediated process that involves immune complexes, antineutrophil antibodies, and abnormal white cell kinetics. We prescribed salazosulfapyridine to a 65-year-old woman with this syndrome. The neutropenia improved along with a reduction in neutrophil-bound IgG, demonstrated by flow cytometric analysis. Salazosulfapyridine may be of benefit for the treatment of Felty's syndrome, and flow cytometry can be used to monitor disease activity and therapeutic efficacy. PMID- 12563704 TI - Infliximab in the treatment of an HIV positive patient with Reiter's syndrome. AB - Reiter's syndrome is an acute inflammatory arthritis with no standard treatment options for patients unresponsive to nonsteroidal antiinflammatory drugs (NSAID). In patients positive for human immunodeficiency virus (HIV), HIV-RNA levels have been correlated with elevated tumor necrosis factor-alpha (TNF-alpha) levels. We investigated the safety and activity of infliximab, an anti-TNF-alpha chimeric monoclonal antibody, in the treatment of an HIV positive patient with Reiter's refractory to NSAID therapy. A 41-year-old HIV positive man with Reiter's syndrome was treated with infliximab 300 mg intravenously at Weeks 0, 2, and 6 and then every 6 to 7 weeks thereafter. He presented with severe fatigue, pain, muscle wasting, synovitis of the elbows, wrists and knees, a scaly rash in the groin area, burning during urination, and severe onycholysis on all digits. Laboratory assessment revealed hemoglobin 7.8 g/dl, erythrocyte sedimentation rate (ESR) 152 mm/h, white blood cell count 5700 cells/mm3, and C-reactive protein (CRP) 65.7 mg/dl. HIV viral load on presentation was 1600 quantitative:ultrasensitive (Qn:US) copies/ml, decreased from a maximum of 428,000 Qn:US copies/ml at the start of antiretroviral therapy. After 6 months taking infliximab, all complaints resolved, nails regrew, and the rash cleared. CRP decreased to 0.8 mg/dl and ESR to 22 mm/h. During this 6 month period antiretroviral therapy remained unchanged, and the viral titer remained below 400 Qn:US copies/ml. PMID- 12563705 TI - B cell loss leading to remission in severe systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) pathogenesis is mediated in part by autoantibodies. We describe a patient with central nervous system lupus who developed a loss of B cells with associated hypogammaglobulinemia and sinopulmonary infections requiring intravenous immunoglobulin. The SLE went into complete remission. Of 18 reported patients with SLE developing persistent hypogammaglobulinemia, only 5 patients including ours had a nearly complete loss of circulating B cells. Of those whose SLE and B cell status was reported, 5/5 with B cell loss and 1/10 without B cell loss experienced a durable response of SLE (p = 0.002). These cases illustrate that B cell ablative therapies may have efficacy for SLE. PMID- 12563706 TI - Chronic arsenic poisoning mimicking Gottron's rash. PMID- 12563707 TI - Gout related upper limb cellulitis: an ultrasound study. PMID- 12563708 TI - [The enlarged translabyrinthine removal of large acoustic neuromas: 18 cases report]. AB - OBJECTIVE: To investigate the surgical methods and results of enlarged translabyrinthine approach in the removal of large acoustic neuromas. METHOD: A large mastoidectomy involved complete exposure of sigmoid sinus, the dura behind the sinus for at least 1 cm, the superior petrosal sinus and the middle fossa dura. The jugular bulb was exposed and pressed downwards if necessary. The internal auditory meatus was skeletonized and uncovered for at least 270 degrees. The debulking of the tumor began inside the anterior and inferior poles in order to find the brainstem and the facial nerve root as early as possible, then the dissection of the nerve was performed medially to laterally. Intraoperative facial nerve monitoring and postoperative CT and MRI were performed in all cases. RESULT: Total removal was achieved in all 18 patients with tumors larger than 3 cm (mean size: 4.2 cm). There were no death as well as other complications such as intracranial infection and persistent cerebrospinal fluid leakage. There were no obvious cerebral sequela. The facial nerve was preserved both anatomically and functionally in 14 cases, with Grade I or II in 8 cases, Grade III or IV in 6 cases. Nerve interruption occurred in 4 cases who all had severe facial paralysis or nerve interruption before operation. 16 patients returned to work within 1-3 months. CONCLUSION: Total removal of large acoustic neuroma could be accomplished via the translabyrinthine approach, with good result of facial nerve function and minimum incidence of morbidity. PMID- 12563709 TI - [Evaluation of the effect of streptomycin perfusion of the labyrinth in the treatment of Meniere's disease and endolymphatic hydrops]. AB - OBJECTIVE: To investigate the clinical effect of streptomycin perfusion of the labyrinth (SPL) in the treatment of Meniere's disease. METHOD: 13 patients with Meniere's disease and 3 patients with delayed endolymphatic hydrops (DEH) underwent SPL. The follow-up time was between 5 and 7 years (average 6.3 years). RESULT: Vertigo was completely controlled in 9 patients, substantialy controlled in 4, limitedly controlled in 2 and not controlled in 1. Hearing was improved in 1 patients, stabilized in 5 and worse in 10. Tinnitus was improved in 3 patients, stabilized in 5 and worse in 8. CONCLUSION: SPL is a safe and effective method in the treatment of Meniere's disease and DEH, but the hearing may be insulted and tinnitus may be worse in some patients. PMID- 12563710 TI - [Clinical observe of the middle ear malformation]. AB - OBJECTIVE: To analyze the diagnosis and therapeutic effect of the middle ear abnormalities. METHOD: 65 ears from 1967 to 1996 were reviewed. The degrees of the ossicular abnormalities were analyzed, operative effect were valued. RESULT: In the 65 ears, incus abnormalities was 57 ears (87.7%), stapes was 55 ears (84.6%), mixed malformation was dominant. 47 ears were operated with ossicular chain reconstruction. 44 ears (93.6%) had hearing improvement, 32 ears (68.1%) had obviously improvement (> 20 dB). They were followed up over one year, 44 ears had steadily hearing and had not any complications. CONCLUSION: If diagnosis and interference were all proper, operation would obtain good result. PMID- 12563711 TI - [Reconstruction for posterior wall of external meatus and sound transmission apparatus after radical mastoidectomy]. AB - OBJECTIVE: To reconstruct middle ear structure for open mastoid antrum with external auditory canal after radical mastoidectomy in one-stage. METHOD: 71 ears of post-mastoidectomy (discharging 53 ears and dried up 18 ears) were undergone with reconstruction of middle ear. The posterior wall of external auditory canal, mastoid cavity and chain of ossicles were reconstructed with homologous costal cartilage. RESULT: 69 ears of 71 cases were near normal structure followed up 6 months to 5 years after operations. The result showed hearing improvement over 15 dB were 55 ears (77.5%) and under 15 dB were 11 cases (15.5%). Five cases (7.0%) were failed. CONCLUSION: Reconstruction of middle ear with homologous costal cartilage is a ideal surgery to reconstruct hearing structure and avoid infection of middle cavity after radical mastoidectomy. PMID- 12563712 TI - [Application of autotransfusion hemodilution in laryngectomy]. AB - OBJECTIVE: To study the application and clinical significance of autotransfusion hemodilution in laryngectomy. METHOD: Autotransfusion hemodilution was adopted in 30 patients with III or IV stage laryngeal carcinoma during operation as the experimental group, and the blood transfusion only was adopted in the patients with excessively blood loss. The other 30 patients with the same stages of disease and the same kinds of operation were performed as the control group, without autotransfusion hemodilution and the blood transfusion was adopted if necessary. RESULT: Blood transfusion was necessary to be performed only in 8 patients of the experimental group, on the contrary, it was necessary in 19 of 30 patients of the control group (P < 0.01). There were no complications during the operations, and the postoperative complications were no significance (P > 0.05) between two groups. CONCLUSION: The application of autotransfusion hemodilution may be of help to a plenty of patients with laryngeal carcinoma in advanced stage to keep off the blood transfusion during laryngectomy. PMID- 12563713 TI - [Expression of CD44s and CD44v6 in laryngeal squamous cell carcinoma and their significance]. AB - OBJECTIVE: To study the correlation between the expression of CD44s, CD44v6 and the clinicopathological characters of laryngeal squamous cell carcinoma (LSCC) so as to analyze the role of them in occurrence and progression of LSCC. METHOD: Expression of CD44 and CD44v6 in 46 cases of LSCC and 20 cases of adjacent normal tissues was inspected with immunohistochemical SP method. RESULT: The expression of CD44s increased significantly in lymphnode metastasis group (94.4%) and stage III-IV group (96.2%), but decreased in non-lymphnode metastasis group (67.9%) and stage I-II group (55.0%). The expression of CD44v6 was lower (21.7%), which wasn't associated with any clinicopathological characters. CONCLUSION: The role of CD44 and CD44v6 in carcinoma maybe dependent on the species, type of carcinoma, and the expression of CD44s may be a biologic marker to evaluate metastasis of LSCC. PMID- 12563714 TI - [The evaluation of cervical lymph node metastasis of laryngeal cancer using magnetic resonance imaging (MRI)]. AB - OBJECTIVE: To assess the potential or limitation of MRI for demonstrating the pathological cervical lymphnodes. METHOD: A prospective diagnostic study on cervical lymphnode metastasis from laryngeal carcinoma was performed for 19 cases undergone neck dissection (5 cases undergone bilateral neck dissection, 24 specimens of neck dissection were collected in total). With pathological findings as the criterion, sensitivity and specificity and accuracy were calculated for palpation and MRI examination of all patients. RESULT: MRI had obviously higher sensitivity specificity and accuracy than palpation. Besides the changes in size and shape of the metastatic lymphnodes, the intensity of MRI inside the metastatic lymphnodes was showed as mixed hypo-isointensity in T1WI and hyper intensity in T2WI. CONCLUSION: MRI examination was accurate in detecting cervical lymphnode metastasis and could image occult lymphnodes which are inaccessible on palpation. Thus, MRI will probably play an important role in the evaluation of malignant lymphnode metastasis. PMID- 12563715 TI - [Nasopharyngeal submucous carcinoma with the first show of cervical lymphnode swell]. AB - OBJECTIVE: To understand the nasopharyngeal submucous carcinoma with the first show of cervical lymph node swell. METHOD: To collect the clinical data of 4 patients with such disease. RESULT: The 4 cases of nasopharyngeal submucous carcinoma were not diagnosed until 3-6 months after cervical lymphnode metastasis. All patients received radiotherapy, three died within 1 year and another survived. CONCLUSION: If some patients have cervical lymph node swell, pay attention to the metastasis of nasopharyngeal submucous carcinoma firstly. Even the examinations such as cytologic test, VCA-IgA, CT, MRI were negative, or even tuberculomas had been diagnosed, nasopharyngeal submucous carcinoma still can not be excluded. PMID- 12563716 TI - [Detection of Epstein-Barr virus DNA in fine needle aspiration specimen from cervical lymphnodes with polymerase chain reaction]. AB - OBJECTIVE: For the diagnosis of occult nasopharyngeal carcinoma. METHOD: Epstein Barr virus in 58 fine needle aspirations from cervical metastatic lymphnodes were detected with polymerase chain reaction. RESULT: The positive reaction was found in 28/35 metastatic carcinoma samples from middle or upside neck. Negative reaction was found in 3 lymphomas, 4 metastatic carcinomas in clavicular superior lymphnodes and 15/16 inflammatory diseases of cervical lymphnodes. The sensibility of this method for the diagnosis of nasopharyngeal carcinoma was 89.3%. The specificity was 86.7%. CONCLUSION: The detection of Epstein-Barr virus in metastatic carcinoma from cervical lymphnodes had some clinical values for the diagnosis of occult nasopharyngeal carcinoma. PMID- 12563717 TI - [Radiofrequency treatment of hemangioma of nasal cavity under nasal endoscopy]. AB - OBJECTIVE: To further understand the advantage of radiofrequency treatment of hemangioma of nasal cavity under nasal endoscopy. METHOD: Fifteen cases with hemangioma of nasal cavity were treated with radiofrequency under nasal endoscopy. RESULT: The hemangioma of fifteen cases could be removed completely. The complication was absent. None of them recurred in six months to four years following up. CONCLUSION: This method has many advantages such as clear operation visual field, less hemorrhage and postoperative pain, no facial scar. The radiofrequency under endoscopy is valuable in treatment for hemangioma of nasal cavity. PMID- 12563718 TI - [The first branchial fistula: an analysis of 16 cases]. AB - OBJECTIVE: To discuss how to improve the level of diagnosis and treatment about the first branchial fistula. METHOD: 16 cases with the first branchial fistula were analyzed retrospectively. RESULT: Of all the patients underwent surgery, 14 cases were free from disease postoperatively within 0.5 years follow-up, 2 patients underwent re-operation because of recurrence. CONCLUSION: Knowing about the disease fully, selecting proper surgical incision and possessing skilled surgical technique is important to improve the level of diagnosis and treatment about the first branchial fistula. PMID- 12563719 TI - [Stereociliary cross-links on the guinea pig utricle hair cells]. AB - OBJECTIVE: To observe the cross-links between stereocilia on guinea pig utricle hair cells. METHOD: Twenty-four utricles from pigmented guinea pigs were examined using high resolution scanning (SEM) and transmission electron microscopy (TEM). RESULT: An extensive network of cross-links occurs between stereocilia of utricle hair cells. These links fall into three categories. Upward-pointing links: from the tips of shorter stereocilia to the side of the adjacent taller stereocilia; Side-to-side links: between stereocilia of the same row; Row-to-row links: between adjacent rows. CONCLUSION: The existence of the stereociliary cross-links binds the hair bundles together and may be associated with the transduction process. PMID- 12563720 TI - [Zrytec drops in the treatment of allergy rhinitis in children]. AB - OBJECTIVE: To assess the efficacy and safety of Zyrtec drops which is a patent product of UCB pharma in treatment of allergy rhinitis in children. METHOD: A total of 47 children of both sexes aged between 2 to 6 years with allergy rhinitis (AR) were included in this study, who were randomly selected to be treated with Zyrtec (Cetirizine 2 HCL) drops 5 mg daily for 3 weeks. Investigators made a global evaluation with symptom scores before treatment and once a week for 3 weeks after treatment. RESULT: An improvement was shown in each week after treatment with statistical significance (P < 0.01). Zyrtec drops provided an excellent improvement at the end of the study. Tolerance was good. CONCLUSION: For acting fast, potent, good tolerance and convenience in use, Zyrtec drops is a better treatment in AR in kids. PMID- 12563721 TI - [Induction of protective immunity in mice against Schistosoma japonicum by nucleic acid vaccine encoding the full-length paramyosin]. AB - AIM: To investigate the immune efficacy of nucleic acid vaccination in mice against full-length paramyosin of Chinese Schistosoma japonicum. METHODS: C57BL/6 and BALB/c mice were vaccinated intramuscularly with the nucleic acid vaccine (pCMV-SjC97) encoding the full-length gene of paramyosin of Chinese S. japonicum. Each group was immunized three times at weeks 0, 3 and 6. Mice vaccinated with pCMV blank vector served as negative control. Mice were challenged three weeks after final DNA boosting by percutaneous infection with cercariae. Six weeks after infection the mice were perfused, worm burden and eggs in the livers, spleens and intestines were counted. Sera from vaccinated mice were collected from the tail vein at weeks 0, 3, 6 and 9, respectively. RESULTS: C57BL/6 mice vaccinated with pCMV-SjC97 produced predominantly IgG2a and IgG2b; whereas in BALB/c mice, IgG1, IgG2a and IgG2b antibodies. Immunization with the pCMV-SjC97 in C57BL/6 mice could confer significant worm reduction rate (35.5%-41.4%, P < 0.05) and egg reduction rate (liver: 44.5%-59.6%, P < 0.05; spleen: 56.7%-82.4%, P < 0.05; intestines: 57.9%, P < 0.05), but not in BALB/c mice. CONCLUSION: The nucleic acid vaccine, pCMV-SjC97, could induce protective immunity in C57BL/6 mice significantly. PMID- 12563722 TI - [Studies on immune responses in mice to expressed products from CSP gene of Plasmodium falciparum FCC1/HN isolate]. AB - AIM: To construct the eukaryotic expression system with pcDNA3-PfCSP/HeLa for CSP gene of Plasmodium falciparum, and to observe the immune responses in BALB/c mice induced by the expressed proteins. METHODS: The recombinant plasmid pcDNA3-PfCSP was transformed into mammalian cell line of human HeLa cells. The expressed protein was isolated and analyzed by SDS-PAGE and used for immunization of BALB/c mice by subcutaneous, intravenous or intraperitoneal administration, respectively. ELISA, Western blotting, T lymphocyte proliferation test, natural killer cell activity and CD4+ and CD8+ T cells detection were used for observation of the level of humoral and cellular immune responses. RESULTS: Immune sera strongly reacted with the expressed protein. The titer of the antibodies was up to 1:6,400 by ELISA. Western blotting analysis revealed a specific band at 38.3 kDa. The specific proliferative response with immunized BALB/c mice spleen cells was remarkablely higher than that with control ones. The ratio of CD4+ and CD8+ T cells and the NK cell activity were significantly higher in the immunization group than that in the control groups. CONCLUSION: The humoral and cell-mediated immune responses and elevated NKC activity to expressed products with the eukaryotic expression system can specifically be detected in BALB/c mice to generate, indicating that the expressed protein could enhance immune activity in mice. PMID- 12563723 TI - [Effects of IFN-gamma gene-modified hepatocytes on TGF-beta 1 and its receptor in mice infected with Schistosoma japonicum]. AB - AIM: To explore the anti-schistosomal hepatic fibrosis effect and the changes in transforming growth factor-beta 1 (TGF-beta 1) and its receptors (TGF-beta RII) in S. japonicum infected mice after intrasplenic transplantation of gamma interferon (IFN-gamma) gene-modified hepatocytes. METHODS: At 16 wk after infection with cercariae of Schistosoma japonicum, the mice were intrasplenically transplantated with murine hepatocytes which had been transfected with IFN-gamma gene-combinant adenovirus vector. ELISA, immunohistochemical and dot blot techniques were used to observe the dynamic changes in IFN-gamma, TGF-beta 1, TGF beta RII and type I, III collagen. RESULTS: The intrasplenic transplantation of IFN-gamma gene modified hepatocytes effectively expressed IFN-gamma and obviously reduced the production and deposition of type I, III collagen as well as TGF-beta 1 and TGF-beta RII. CONCLUSION: IFN-gamma gene transplantation has anti-hepatic fibrosis efficacy in Schistosoma japonicum-infected mice, being related to its role of decreasing the expression of TGF-beta 1 and TGF-beta RII. PMID- 12563724 TI - [A case of ancylostomiasis manifested by fresh bloody stool]. PMID- 12563725 TI - [Immune responses in mice vaccinated with recombinant plasmid pcDNA3 containing ROP1 gene from Toxoplasma gondii]. AB - AIM: To observe the immune responses in BALB/c mice vaccinated with the constructed recombinant plasmid, pcDNA3, containing ROP1 gene from Toxoplasma gondii. METHODS: The plasmid DNA obtained by alkaline lysis were injected into the left leg of each mouse at a dosage of 100 micrograms. A booster vaccine was given using the same dosage two weeks later. Control groups were injected with the pcDNA3 blank plasmid and normal saline, respectively. 30, 50 and 70 days after the booster injection, the proliferation activity of T lymphocytes and the NK cell activity were determined using MTT assay, the number of CD4+/CD8+ T cells with indirect immunofluorescence assay and the titer of IgG antibody by ELISA. RESULTS: The spleens of mice enlarged obviously after immunization with pcDNA3 ROP1. The activity of proliferation of spleen T lymphocytes was higher in the immunization group than in the control groups; The NK cell activity in the immunization group was higher than those of control ones. The number of CD4+ T cells showed no obvious increase but CD8+ T cells obviously increased. The titer of IgG antibody in the immunization group showed no significant increase until 90 days later. CONCLUSION: Vaccination of mice with the recombinant plasmid, pcDNA3 ROP1, could elicit cellular and humoral immune response in mice. PMID- 12563726 TI - [Misdiagnosis of 4 cases of amoebic dysentery]. PMID- 12563727 TI - [Establishment and identification of the monoepitopic monoclonal antibody of Plasmodium falciparum]. AB - AIM: To prepare a monoepitopic monclonal antibody of Plasmoduim falciparum. METHODS: According to the theory of protein structure, a peptide with 6-9 residues representting the antigenicity of the original histidine-rich protein II (HRP-II) of P. falciparum was synthesized and used to immunize BALB/c mice after purity identification by capillary electrophoresis. RESULTS: Four hybridoma cell lines targeting a suitable peptide from pf HRP-II were obtained by using spleen embedment method and hybridoma technology. CONCLUSION: It is the first report to prepare a monoepitopic monoclonal antibody against original protein by selecting a suitable peptide from the primary sequence of protein. PMID- 12563728 TI - [Two misdiagnosed cases of cerebral schistosomiasis]. PMID- 12563729 TI - [Effect of triclabendazole on the ultrastructure of body wall and vitelline cells of Paragonimus westermani]. AB - AIM: To observe the ultrastructural changes in the body wall and the vitelline cells of Paragonimus westermani in vitro and in vivo before and after triclabendazole treatment. METHODS: The worms were obtained from in vitro and in vivo tests. All of the samples were processed by conventional techniques, and observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). RESULTS: The external plasma membrane and matrix were cracked or disappeared after the treatment. The necrosis of the muscular layer differed. The cell membranes of cortex and vitelline cells were damaged. Nuclear membrane was damaged partially, heterochromatin solidified and condensed to brim and dissolved. The Golgi complex disappeared, endoplasmic reticulum expanded, mitochodria denatured and dissolved. The damage was more serious in vivo than in vitro. CONCLUSION: Triclabendazole is remarkablely effective against Paragonimus westermani by damaging the body wall and vitelline cells, mainly affecting the nuclei, membrane structures and microtubular system. PMID- 12563730 TI - [Analysis of kinetoplast DNA of Leishmania isolates in China by PCR-SSCP]. AB - AIM: To analyse the kDNA of the pathogens of leishmaniasis in China. METHODS: Based on leishmaniasis specific primers 13A, 13B and a set of oligonucleotide primers I and II with Leishmania donovani (L. d.) Sichuan isolate specificity, PCR were conducted to amplity minicircle kDNA fragments (297 bp and 120 bp) in the pathogens of leishmaniasis from different epidemiologic foci in China. The products were analyzed by single strand conformation polymorphism technology (SSCP). RESULTS: PCR amplified 297 bp product occurred in L. d. isolates from hill and desert foci, but no product was found in L. d. isolates from plain foci in China. SSCP of these 297 bp kDNA fragments showed that there was no difference in the mobility of ssDNA between isolates from hill foci, but there was apparent difference in the mobility of ssDNA between L. d. isolates from hill and desert foci. PCR amplified 120 bp products occurred in L. d. Sichuan isolate, L. d. Wenchuan isolate, L. d. Gansu isolate from hill foci and L. d. Shandong isolate and L. d. Jiangsu isolate from plain foci. SSCP of the 120 bp products showed that no difference in the mobility of ssDNA was found between two isolates from plain foci. There was also no difference in the mobility of ssDNA between L. d. Wenchuan isolate and L. d. Gansu isolates from hill foci. But there was apparent difference in the mobility of ssDNA between L. in fantum and L. d. isolates from different foci. CONCLUSION: Heterogeneity does exist between the kDNA of L. d. isolates from different foci of leishmaniasis of China. PMID- 12563731 TI - [A case of myiasis of the knee joint caused by Musca domestica vicina]. PMID- 12563732 TI - [Anti-Toxoplasma effect of activated mouse macrophages induced by interferon gamma combined with TNF-alpha]. AB - AIM: To compare the inhibitory effect of the macrophages activated by IFN-gamma combined with TNF-alpha against RH strain and Fukaya strain. METHODS: The average parasite proliferation rates of the two strains within the cytokine-activated M phi s were calculated at different times post-challenge, the nitric oxide (NO) levels in the medium supernatant were simultaneously determined. RESULTS: In the macrophages activated by 100 U each of IFN-gamma and TNF-alpha, the invaded tachyzoites of RH strain were completely killed, while the invaded tachyzoites of Fukaya strain remained slow proliferation with significantly lower levels of NO detected at 24 h post challenge. CONCLUSION: The difference in the anti Toxoplasma effect of the activated macrophages against RH and Fukaya strains might be attributed to the different amount of NO produced by the macrophages. PMID- 12563733 TI - [Comparison of sensitivity of artesunate-sensitive and artesunate-resistant Plasmodium falciparum to chloroquine and amodiaquine]. AB - AIM: To explore the in vitro sensitivity of artesunate-sensitive and -resistant Plasmodium falciparum to chloroquine and amodiaquine and to observe the effect of artesunate combined with chloroquine and artesunate combined with amodiaquine on artesunate-resistant P. falciparum. METHODS: The sensitivity of the artesunate sensitive and -resistant P. falciparum to chloroquine and amodiaquine and their combination with artesunate was compared by using Rieckmann's in vitro micro technique. RESULTS: The ID50 and ID95 values of chloroquine, amodiaquine and artesunete were 90.9, 50.9, 9.6 nmol/L and 320.0, 320.0, 40.0 nmol/L to the artesunate-sensitive P. falciparum respectively and were 112.0, 133.5, 85.1 nmol/L and 320.0, 320.0, 400.0 nmol/L to artesunate-resistant P. falciparum, respectively. When artesunate was combined with chloroquine, the ID50 values of the 2 drugs were 3.2 and 20.0 nmol/L to the artesunate-resistant P. falciparum. When artesunate was combined with amodiaquine, the ID95 values of the 2 drugs were 3.2 and 5.0 nmol/L to the artesunate-resistant P. falciparum, respectively. CONCLUSION: The artesunate-resistant P. falciparum has no cross resistance to chloroquine and amodiaquine, however, artesunate combined with chloroquine or amodiaquine exhibit an apparent synergistic effect in vitro. PMID- 12563734 TI - [Detection of Plasmodium vivax by polymerase chain reaction in Hainan]. AB - AIM: To establish a modified polymerase chain reaction(PCR) method for the detection of Plasmodium vivax in the endemic areas of malaria and compare the method with the conventional light microscopy in the field. METHODS: A PCR method was modified by improving the collection procedures of blood samples, template extraction, primer design and optimizing the reaction condition. The method was evaluated by examining blood samples from 310 patients with vivax malaria and compared with the conventional light microscopy in endemic areas of Hainan Province. RESULTS: The positive rates of the modified PCR method and microscopic method were 34.2% and 31.9%, respectively. CONCLUSION: The modified PCR method is simple, sensitive and specific for the detection of vivax malaria patients in endemic areas. PMID- 12563735 TI - Expression of Plasmodium falciparum-infected erythrocyte membrane protein from cerebral malaria patients. AB - AIM: To provide theoretical evidence for studying the molecular pathogenesis of human cerebral malaria. METHODS: The expressions of Plasmodium falciparum erythrocyte membrane protein 1(PfEMP1) on the surface of parasitized erythrocyte (PE) specimens from 19 cases of cerebral malaria patients in Yunnan Province were quantitatively analyzed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) technique. 43 patients of falciparum malaria, 9 patients of vivax malaria and 6 healthy controls were also investigated. RESULTS: The expressions of higher molecular mass (Mr) 260-320 kDa forms of PfEMP1 were found on PE from cerebral malaria patients. By contrast, the expression of PfEMP1 and P. vivax erythrocyte membrane protein (PvEMP1) on PE from falciparum malaria patients and vivax malaria patients had a PfEMP1 with Mr 240 kDa and a PvEMP1 with Mr 180 kDa band, respectively. Healthy controls expressed an EMP of Mr 140 kDa. CONCLUSION: The binding of 260-320 kDa PfEMP1 proteins expressed on PE from cerebral malaria patients to diverse receptor molecules on the endothelial cell(EC) of the cerebral microvessels such as CD36, thrombospondin (TSP), intercellular adhesion molecule 1(ICAM-1), vascular cell adhesion molecule 1(VCAM 1), endothelial leukocyte adhesion molecule 1(ELAM-1) and chondroitin sulfate A (CSA) might be the molecular basis for the pathogenesis of cerebral malaria. PMID- 12563736 TI - [Finding of new FCC1/HN antigenic expressed sequence tag(ESTs) of Plasmodium falciparum]. AB - AIM: To sequence the strong positive clones obtained by immuno-screening of Plasmodium falciparum FCC1/HN lambda gt11 cDNA expression library, and to elucidate the antigenic expressed sequence tags through sequencing the cDNA insert of these positive clones, and new antigenic ESTs could serve as a resource to pursue their corresponding antigen genes. METHODS: cDNA inserts of positive lambda gt11 phage clones were amplified by PCR. The PCR products, after purification, were cloned into the M13 mp18 sequencing vector. Single-stranded M13 DNA was prepared and sequenced. Then the acquired sequences were compared in homologies with EMBL/GenBank database on the PC/GENE software system and searched in NCBI (National Center for Biotechnology Information) GenBank using BLAST (Basic Local Alignment Search Tool) commond. RESULTS: Sequence C03 was part of the known P. falciparum antigenic heat shock protein 70 (Pfhsp70) gene, while the other 5 sequences were new P. falciparum antigenic expressed sequence tags (ESTs). CONCLUSION: The 5 new antigenic ESTs generated could serve as the breaking through points in our efforts to find out new P. falciparum antigen genes. PMID- 12563737 TI - [SDS-PAGE and EITB analysis of the protein components of different isolates of Schistosoma japonicum in China and Japan]. AB - AIM: To make a supplementary observation on the protein components prepared from adult S. japonicum of 4 different isolates from Zhejiang, Jiangxi, Taiwan Provinces of China and Japan in origin and to observe antigen reactivity of the above-mentioned isolates together with adult worms from Anhwi, Hubei, Sichuan and Yunnan isolates against heterologous anti-Oncomelania h. hupensis (collected from Guangxi, China and Japan) sera by EITB. METHODS: SDS-PAGE and EITB. RESULTS AND CONCLUSION: SDS-PAGE showed that by Coomassie blue staining, male S. japonicum from Jiangxi, Zhejiang, Taiwan Provinces and Japan isolales revealed 7-17 bands while female worms revealed 1-6 bands. The protein patterns of Taiwan and Zhejiang male worm were similar, but slight difference could be seen above 81 kDa. By silver staining, male worms of the 4 isolates revealed 10-23 bands while female worms revealed 1-19 bands. Male worms from Japan not only showed less bands but also differed in their pattern as compared to those of other Chinese isolates. Results of EITB revealed that each of the 4 isolates had slightly different pattern but all of the tested isolates had common antigens with their heterologous snail hosts i.e., Oncomelania h. hupensis from Guangxi, China and Japan. PMID- 12563738 TI - [Biological characteristics of the juvenile Paragonimus westermani]. AB - AIM: To observe the biological characteristics of the juvenile of Paragonimus westermani. METHODS: Kunmin strain mice and Wistar rats inoculated orally with 100-150 metacercariae of Paragonimus westermani were autopsied 10-30 days and 80 580 days after infection, respectively. Life-span, survival time in dead hosts or physiological saline at 5 degrees C to 8 degrees C and invasiveness of juveniles to new hosts were detected by host transfer. RESULTS: All the worms recovered from the mice and rats were stunted juveniles. The worm detection rates in the mice and rats were 19.3% and 22.2%-37.5%, respectively. The life-span of the juveniles was rather long, being not limited by the life-span of a host. Host transfer could prolong longevity and preserve invasiveness. The juveniles recovered from dead mice still possess strong vitality and invasiveness. CONCLUSION: The juvenile of P. westermani is an infective stage, having strong vitality and invasiveness. PMID- 12563739 TI - [Surveillance of imported falciparum malaria in Yongkang city, Zhejiang Province in 1985-1998]. PMID- 12563741 TI - [Comparison of the efficacy of albendazole-medicated sweets and praziquantel for the treatment of clonorchiasis]. PMID- 12563740 TI - [Immunoregulation of IL-2 in Trichinella-infected mice]. AB - AIM: To study the immunoregulation of IL-2 in Trichinella-infected mice. METHODS: Mice infected respectively with HL strain and AM strain larvae of Trichinella spiralis were treated i.p. with IL-2 from the second day postinfection for 3 days. Serum IgG antibody levels were determined by ELISA and the infection capacity was determined using reproductive capacity index (RCI). RESULTS: In HL, higher dosage of IL-2 injection induced lower RCI and showed apparent anti Trichinella effect. In AM, both low and high dose of IL-2 had no measurable effect on RCI, however, high dose of IL-2 reduced the infectivity of newborn larvae. CONCLUSION: IL-2 exhibits apparent suppressive effect on the infectivity of T. spiralis of HL strain. PMID- 12563742 TI - [Dynamic changes in IL-4, IL-5 and IL-10 in liver and bone marrow of mice infected with Schistosoma japonicum]. AB - AIM: To observe the changes in Th2 cytokines in the liver and bone marrow of mice infected with schistosomiasis japonica. METHODS: ABC immunohistochemical staining technique and multimedia pathological picture analysis were used. Meanwhile, IL 4, IL-5 and IL-10 derived from livers and bone marrows of infected mice were observed at wk 8, 10 and 12 after infection. RESULTS AND CONCLUSION: In the livers of infected mice, the levels of IL-4, IL-5 and IL-10 increased obviously with a prolongation of the infection duration, IL-4 being the highest. However, in the bone marrow of infected mice, IL-4 level increased slowly with the duration of the infection, being lower than those in the liver at wk 10 and 12 after infection. The level of IL-5 was higher than that in the liver within 12 wk. At wk 10 after infection IL-10 tended to increase, but decreased thereafter and was obviously lower than that in the liver. In mice infected with Schistosoma japonicum, the immune response occurred mainly in the liver. IL-5 in the bone marrow showed auto-secretion. PMID- 12563743 TI - [Studies on detection of Plasmodium falciparum and Plasmodium vivax in blood samples by multiplex polymerase chain reaction]. AB - AIM: To establish a sensitive and specific PCR-based method to detect Plasmodium falciparum and P. vivax in blood samples in a single amplification reaction. METHODS: Malaria parasite DNA in blood was amplified by the multiplex polymerase chain reaction using two sets of primers derived from the P. f. moderately repetitive DNA sequence and COIII gene of P.v. RESULTS: A 206-bp product for P. f. and a 370-bp product for P.v. were amplified by multiplex PCR, being able to detect parasitemia level as low as 5 x 10(-7) for P. f. and 1.02 x 10(-6) for P. v. and having no cross-reaction with human leucocyte DNA. A total of 783 blood samples on the filter paper collected from patients attending to malaria clinics in malaria endemic areas were detected. The positive rate of multiplex PCR was 85.8%, the misdiagnosis rate was 0, and the under-diagnosis rate was 0.1%, while these three rates of microscopic examination were 84.9%, 3.1% and 1.0%, respectively. The concordance between the two methods was 95.8%. CONCLUSION: The multiplex PCR method made the malaria detection more sensitive and specific than the microscopic examination and should be suitable for the diagnosis of malaria in mixed endemic areas, large-scale epidemiological studies, follow-up of drug treatment and donor blood screening. PMID- 12563744 TI - [Cellular immune response in mice vaccinated with uv-attenuated Toxoplasma gondii ZS1 strain trophozoites]. AB - AIM: To explore the protective effect and cellular immune response of uv attenuated ZS1 strain trophozoites of Toxoplasma gondii in mice. METHODS: The ZS1 strain trophozoites of T. gondii were irradiated by uv-light with 2537 A wave length for 60 minutes. Mice were divided into 4 groups. Group 1 was vaccinated alone, group 2 was challenged with normal ZS1 trophozoites on d45 after vaccination, group 3 was infected alone, and group 4 was normal control. The changes in splenic T cell proliferation, level of CD4+ and CD8+ T cell, and NK cell activity were compared. RESULTS: Group 1 mice survived normally, no trophozoite, pseudo-cyst or cyst was detected in the tissues on d49 after vaccination. Group 2 mice survived longer than those of group 3. The T lymphocyte proliferation in response to soluble antigen of T. gondii was significantly enhanced in group 2, and suppressed in group 3. The level of CD4+ T cell in group 2 was decreased, resulting in a reverse of CD4+/CD8+ ratio. The NK cell activities in groups 1, 2 and 3 were all significantly increased. CONCLUSION: The uv-attenuated vaccine of T. gondii ZS1 strain could induce certain protective immunity against challenge infection, in which CD8+ T cell and NK cell might play an important role. PMID- 12563745 TI - [The protective effect against Toxoplasma infection in mice immunized with laser irradiated Toxoplasma tachyzoites]. AB - AIM: To observe the level of immune responses and protective immunity in mice induced by laser-irradiated Toxoplasma tachyzoites (LIT). METHODS: ICR mice were immunized with LIT. After one month, the mice were challenged with live Toxoplasma tachyzoites to observe the effect of LIT. Indirect immunofluorescence assay and ELISA were used to detect CD4+/CD8+ lymphocyte subpopulation and IgG antibody. RESULTS: Immunization with LIT could partially protect the mice from Toxoplasma infection, prolong the survival time, enhance the CD4+/CD8+ ratio and produce specific IgG antibody. CONCLUSION: Intraperitoneal injection with LIT can induce partial protective immunity and specifically elicit ICR mice to generate humoral and cell-mediated immune responses. PMID- 12563747 TI - [Resource for malaria research information on Internet]. PMID- 12563748 TI - [Comparison of two stool examination methods for the diagnosis of clonorchiasis]. PMID- 12563749 TI - [Dynamic CT observation of 5 cases of cerebral paragonimiasis]. PMID- 12563746 TI - [Changes in computed tomogram in cerebral parenchymal cysticercosis treated with albendazole]. AB - AIM: To observe the changes in cerebral computed tomogram (CT) in cerebral parenchymal cysticercosis after albendazole treatment. METHODS: Cerebral CT scanning was conducted in 57 patients with cerebral cysticercosis in our hospital before, during and after albendazole treatment. RESULTS: Cerebral CT might be normal before onchospheres became cysticercariae in the brain. Small cystic lesions could turn into other CT signs of cerebral cysticercosis after albendazole treatment. CONCLUSION: Normal cerebral CT cannot rule out the disease. Small cystic lesions are the earliest pathological and active signs of the disease. Nodular focus occurs after the death of Cysticercus and calcification is the final outcome of the disease. PMID- 12563750 TI - [Survey of the relationship between hookworm disease and socioeconomic factors]. PMID- 12563751 TI - [Laboratory rearing technique of Anopheles sinensis]. PMID- 12563752 TI - [Clinical analysis of 68 newborns with malaria in Mali]. PMID- 12563753 TI - [A survey of clonorchiasis in the New Zhuzhao River valley, Shandong Province]. PMID- 12563754 TI - [Malaria situation in the People's Republic of China in 1998]. AB - Notwithstanding there was a flood over the five provinces located at the middle and lower reaches of the Changjiang River, i.e. Hunan, Hubei, Jiangxi, Anhui and Jiangsu, in the summer and autumn of 1998, the prevalence of malaria in these areas was basically stable or slightly decreased as a result of timely implementing active and effective preventive measures. According to the case reporting system, the number of malaria cases of the whole nation totaled 31,319 in 1998, with a mean incidence of 2.53 per a hundred thousand, and 24 deaths. A decrease of 12.0% in incidence was revealed as compared with that in 1997. The county-based reckoning showed that absence of malaria cases or drop of the incidence to lower than 1@10000 was reported from areas in 2,774 counties with a coverage of 1.2016 billion people; areas with 30.6 million people distributed in 75 counties showed an incidence of 1.1-10@10000, and areas with 6.50 million people distributed in 27 counties showed an incidence of 10.1@10000-100@10000. Of the 17 provinces, autonomous regions and municipalities (P/A/M) where major malarious areas are located, 6 showed more or less increase in malaria incidence, particularly noteworthy for Hainan and Yunnan Provinces, where 5,043 and 12,988 malaria cases were reported respectively, accounting for 57.6% of the national total. However, the actual number of malaria cases of the two provinces was estimated to be ten folds the number of the reported cases. Malaria surveillance was performed in areas covering 1,298 counties in 17 P/A/M with a coverage of 589 million people. Blood examination on febrile patients for 8,712,454 man/time detected 27,090 cases positive for malaria parasite, showing a mean positive rate of 0.31%, of which the number of falciparum malaria cases was 4,042 (including mixed infections with vivax malaria, the same below), a proportion of 14.9% of the total parasite positives was exhibited. Altogether 597,111 targeted people, inhabitants in epidemic spots and migratory people were also examined, 4,326 people were found to be positive for malaria parasites, showing a mean parasite rate of 0.72%, while falciparum malaria carriers were noted as 748 among them. As a whole, the endemicity of falciparum malaria was still confined to 47 counties of Hainan and Yunnan Provinces. Imported falciparum malaria cases were reported in 84 counties of 15 P/A/M (including non-endemic areas of Hainan and Yunnan), whereas no introduced falciparum malaria cases were recorded. The distribution of malaria did not show conspicuous changes in recent years. The prevalence was most severe in mountainous regions in Hainan with Anopheles dirus as the major vector, followed by areas south to 25 degrees N.L. where An. minimus distributed and areas between 25 degrees N.L. with An. anthropophagus as the vector. The three mentioned anopheline species were distributed in areas where more than 200 million people resided, the number of malaria cases amounted to 90% odd of the national total, the prevalence of malaria was unstable there, and focal outbreaks were often reported. Nevertheless, in areas only distributed with An. sinensis, the endemicity of malaria was stable, and its incidence has been reduced to less than 0.1@10000. PMID- 12563755 TI - [Cloning, sequencing and expression of the full-length gene encoding paramyosin of Schistosoma japonicum in vivo]. AB - AIM: To clone and sequence the gene encoding paramyosin of S. japonicum (Chinese strain) and to study the expression of the DNA-based vaccine encoding the full length paramyosin of S. japonicum in vivo. METHODS: Total RNA was isolated from adult S. japonicum using TRIzol reagent. The full-length cDNA encoding paramyosin of S. japonicum was amplified by RT-PCR and cloned into pGEM-T vector and sequenced by the method of dideoxy-mediated chain-termination. The cDNA encoding paramyosin of S. japonicum was subcloned into the expressive plasmid vector pCDNA/AMP(pCMV-Sjc97), and the recombinants were identified by restriction enzyme digestion and sequencing. The immunofluorescence assay was used to study the expression of Sjc97 in vivo in mice. RESULTS AND CONCLUSION: The 2.6 kb cDNA encoding the full-length paramyosin of Chinese S. japonicum has been successfully cloned and sequenced for the first time. The full-length sequence of paramyosin of S. japonicum was determined. Comparison of the nucleotide sequence and the deduced amino acid sequence of Sjc97 with that of S. japonicum paramyosin (Philippine strain) (Sjp97), S. japonicum paramyosin(Japanese strain) (Sjj97), S. mansoni (Sm97), B6 and Y6 clone (the partial cDNA encoding paramyosin of Chinese strain) showed that Sjc97 differed from Sjp97 by 16/2,601 nucleotide and 3/866 amino acid substitutions (99.4% on nt-level and 99.7% on aa-level in homology); from Sjj97 by 20/2,601 nucleotide and 2/866 amino acid substitutions (99.2% on nt level and 99.8% on aa-level in homology); and from B6 by 11/1,329 nucleotide and 1/443 amino acid substitutions (99.0% on nt-level and 99.8% on aa-level in homology); from Y6 by 13/1,329 nucleotide and 1/443 amino acid substitutions (98.9% on nt-level and 99.8% on aa-level in homology); Sjc97 differed from the Sm97 by 2,235/2,601 nucleotide and 34/866 amino acid (91.0% on nt-level and 96.0% on aa-level in homology). The plasmid expression vector encoding the full-length paramyosin of Chinese S. japonicum has been successfully constructed. The pCMV Sjc97 vaccine could express Sjc97 protein in vivo in mice after intramuscular immunization. PMID- 12563756 TI - [Malaria surveillance in Huzhou city, Zhejiang province in 1990-1998]. PMID- 12563757 TI - [Hybridization experiments using Anopheles minimus from Hainan and Yunnan]. AB - AIM: To observe whether there were any intra-species differences between Anopheles minimus from Hainan(H) and Yunnan(Y). METHODS: Anopheles minimus were collected from cattle shed on the spot. Each isofemale line was set up in the laboratory. Hybridization experiments were conducted by using forced mating between Anopheles minimus from Hainan and Yunnan, for observing the reproductive ability of F1 hybrids. Ovarian nurse cell polytene chromosomes of F1 hybrid females were examined, to observe any synapsis in different zones of chromosomes. RESULTS: No embryo formation was found within the eggs produced by group Y female x H male, the hatching rate was zero. Low hatching rate was shown in other groups with (H female x Y male) F1, except for groups with (H x Y) F1 x Y. Ovarian nurse cell polytene chromosomes from (H x Y) F1 hybrid females showed constant asynapsis at the 29th, 36th and 37th zones in the chromosome 3R, and at the 4th and 6th zones in the chromosome X. CONCLUSION: Reproductive isolation did appear in An. minimus from Hainan and Yunnan. PMID- 12563758 TI - [Experimental study on the contamination of the brook with Paragonimus metacercariae]. PMID- 12563759 TI - [Studies on the feasibility of detecting circulating antibodies in saliva of Schistosoma japonicum infected rabbits]. AB - AIM: To investigate the feasibility of detecting anti-Schistosoma japonicum antibodies in saliva. METHODS: Saliva and serum samples of 5 infected, 7 reinfected and 8 treated rabbits were collected at different times periods. The CAb in saliva and serum was detected by using ELISA. RESULTS: The sensitivity of ELISA was 94.7% for saliva and 100% for serum. The specificity of ELISA was 100% for both saliva and serum. CONCLUSION: Saliva can be used to detect circulating antibodies for the diagnosis of schistosomiasis japonica. PMID- 12563760 TI - [Expression and identification of recombinant 22.6 kDa fusion protein of Schistosoma japonicum]. AB - AIM: To obtain a large amount of purified 22.6 kDa antigen of Schistosoma japonicum (Sj22.6) in large quantity. METHODS: The sequence of the gene fragment encoding Sj22.6 was reformed by PCR and subcloned into plasmid vector pGEX-1 lambda T that coded for the 26 kDa GST antigen of Schistosoma japonicum (Sj26 GST). The recombinant plasmid was transformed into E. coli TG2 and then the positive recombinant clone was expressed by induction with IPTG. RESULTS: The recombinant Sj22.6/Sj26 GST fusion protein was expressed in 5.1% of total bacterial protein and was easy to be purified with glutathione sepharose 4B. Moreover, the purified recombinant Sj22.6 antigen could be cut off easily from the fusion protein with thrombin and had high immunogenicity. CONCLUSION: The purified recombinant Sj22.6 protein and Sj22.6/Sj26 GST fusion protein had the same immunological activity as the native Sj22.6 kDa protein. PMID- 12563761 TI - [A case with amoebic vaginitis]. PMID- 12563762 TI - [Studies on the continuous culture and the growth of Entamoeba gingivalis]. AB - AIM: To establish a method for continuous culture of Entamoeba gingivalis(E. g.). METHODS: The culture conditions of E. g. were compared by observing its size and survival time at room temperature. RESULTS: The growth of E. g. under different culture conditions including culture medium, temperature and pH were compared. The accompanying bacteria associate with E. g. FJ4 were isolated and identified. The average size of E. g. was 13.19 microns-49.93 microns x 9.88 microns-30.74 microns. The optimal culture conditions of E. g. were: modified LES or YES medium, pH 6.4-6.7, nutritional liquids such as Locke's solution or yolk liquid with 20% bovine serum, penicillin, streptomycin and rice flour at 35 degrees C. Reproduction of E. g. peaked at the fourth day of incubation, and the survival time of E. g. was 120 h-168 h. CONCLUSION: E. g. could be continuously cultured in modified LES or YES medium by inoculating once every four days. PMID- 12563763 TI - [Studies on the relationship between the infection of intermediate hosts of Paragonimus and ecological environment]. AB - AIM: To explore the relationship between the infection of the intermediate hosts of Paragonimus and their ecological environment. METHODS: Three different villages in Fujian Province were chosen to conduct the study. The infection of snails and crabs were surveyed. RESULTS: 1. In Ningyang Village, Wuyi mountain(stream type), the infection rate of the Potamon crabs (723.9/each) in section I (rapid stream) was 5 times higher than that in section II (rapid stream) 142.6/each. 2. In Xikou Village, Mingching county(ditch type): The infection rate of the snails (Semisulcospira libertina) to cercaria and the Potamon crabs were 4.2% and 1,738.3/each, respectively in 1996, being 28.9 and 8.5 times higher than those (0.2% and 204.7/each) in 1982. 3. In Jishang Village, Jianou City, the infection rate of the snails (Tricula) and the Potamon crabs were 4.3% and 355.4/each respectively in ditch III, 4.3 and 8.3 times higher than those (1% and 42.9%/each) in ditch II. CONCLUSION: P. westermani distributed along the stream system, usually concentrating in the small, slow-running water bodies of the stream. P. skrjabini distributed in still smaller water bodies. The environment suitable for the breedings of the snails and crabs are also the suitable places for Paragonimus parasites to complete their life cycles. PMID- 12563764 TI - [Forty-one cases of severe malaria treated with novoxacin]. PMID- 12563765 TI - [Fusion expression of porcine IFN-gamma and Cysticercus cellulosae antigen cC1 in Escherichia coli cells]. AB - AIM: To construct an expression vector, including a chimeric cDNA of porcine IFN gamma and Cysticercus cellulosae antigen cC1. METHODS: DNA fragments of porcine IFN-gamma and cC1 including linkers were generated by polymerase chain reaction (PCR). The recombinant vector pJLA-PRcC1 was constructed by inserting a chimeric cDNA of porcine IFN-gamma and Cysticercus cellulosae antigen cC1, and transformed to E. coli XL-Blue. RESULTS: The recombinant vector was identified by restriction analysis. An inserted fragment about 1.5 kb could be found. After induction, a 52 kDa new protein band appeared in SDS-PAGE. CONCLUSION: The fusion expression of porcine IFN-gamma and cC1 antigen in Escherichia coli cells is successful. PMID- 12563766 TI - [A case of congenital cerebral toxoplasmosis]. PMID- 12563768 TI - [Modified method for staining malaria parasite in thick blood smear]. PMID- 12563767 TI - [Expression of Schistosoma japonicun fatty acid binding protein gene in silkworm cells and larvae]. AB - AIM: To express the fatty acid binding protein (Sj14FABP) gene of Schistosoma japonicun in the silkworm cells and larvae. METHODS: A 600 bp DNA fragment containing Sj14FABP gene was cloned into baculovirus transfer vector of pBacPAK His1 to construct recombinant transfer vector Sj14-pBac PAK His1. Coinfection was accomplished with this vector and Bombyx mori nuclear polyhedrosis virus (BmNPV) DNA in BmN cells. The recombinant virus of Bm-Sj14 was screened using dot blotting. The BmN cells and silkworm larvae were infected with Bm-Sj14 to express Sj14FABF gene. Western blotting and ELISA were used to identify the antigenicity of the recombinant protein. RESULTS: Sj14FABP gene was successfully expressed in the BmN cells and silkworm larvae infected with Bm-Sj14. The product was a 18 kDa fusion protein. The yield in BmN cells was about 100 micrograms/1 x 10(6) cells and 33 micrograms/ml cell supernatant. In silkworm larvae, the product yield was 4 mg/ml haemolymph as well as 4.6 mg/g silkworm tissue. The recombinant protein could be recognized by Western blotting and ELISA using the sera from mice immunized with SWAP. CONCLUSION: Sj14FABP gene has been successfully expressed in BmNPV system and the product has high antigenicity. PMID- 12563769 TI - [Immunocytochemical localization of two facilitated glucose transporters of Schistosoma mansoni in the tegument of Schistosoma japonicum]. AB - AIM: To observe the distribution of Schistosoma mansoni glucose transport proteins, SGTP1 and SGTP4, in the tegument of Schistosoma japonicum. METHODS: The rapidly frozen fixation technique and ultracryomicrotomy were adopted for preparing ultrathin cryosections of S. japonicum. Anti-SGTP1 and anti-SGTP4 antibodies were used to localize the corresponding antigens in the tegument of adult S. japonicum by immunocytochemical technique. RESULTS: SGTP1 was localized on the basal membrane of the tegument and its infoldings, SGTP4 was localized on the apical membrane of the tegument and its invaginations of S. japonicum. CONCLUSION: The same localization for SGTP 1 and SGTP4 in the tegument of S. japonicum and S. mansoni exhibited apparent homology between SGTPs of the two schistosomes. PMID- 12563770 TI - Establishment of immunoglobulin M(IgM)-immunosorbent agglutination assay (ISAGA) for diagnosis of toxoplasmosis. AB - AIM: To establish an immunosorbent agglutination assay (ISAGA) for detection of IgM antibodies against Toxoplasma gondii. METHODS: In the ISAGA, wells of microtiter plates were coated with anti-human IgM antibodies and sealed with 1% bovine serum albumin. After the test sera were added and incubated, the plates were washed, T. gondii tachyzoite antigen suspension was added, and incubated overnight at 37 degrees C. The ISAGA results were evaluated by comparing with those detected by Danish ISAGA and ELISA and those detected by slide enzyme immunoassay (S-EIA). RESULTS: Forty-four sera from Danish pregnant women were tested by the IgM-ISAGA, 41(93.2%) were consistent with the Danish results. Sixty seven sera from Danish and Shanghai pregnant women were detected by IgM-IgM-ISAGA and S-EIA, the total consistency rate was 92.5%. A significant correlation was found between the titers of the ISAGA and S-EIA, with a Pearson correlation coefficient of 0.589(P < 0.001). The titers of ISAGA were eighteen times higher than those of S-EIA. This method enables the detection of IgM antibodies as low as approximately 0.08 IU/ml. CONCLUSION: The IgM-ISAGA is therefore sensitive, specific, easy to perform, and is useful for mass screening and diagnosing recent Toxoplasma infection or reactivation. PMID- 12563771 TI - [Isolation and quantitation of chloroquine-binding proteins in Plasmodium berghei]. AB - AIM: To isolate and quantitate chloroquine (CQ)-binding proteins in both CQ sensitive (CS) AND CQ-resistant (CR) Plasmodium berghei. METHODS: P. berghei infected mice were each given i.g. CQ 400 mg/kg, respectively, 3 h later, the proteins of the parasites collected were separated using Ultrogel AcA34 gel column, followed by CQ extraction, the CQ concentration was determined by HPLC (extra standard). RESULTS: There were 58 protein peaks in the CS, all of which were bound with CQ, fraction Nos. 64-86 (peak 9) being the predominant, accounting for 16.4% of the total amount of CQ-binding protein. In the CR there were 40 protein peaks, among which 32 were bound with CQ, the predominant fraction Nos. 60-83 (peaks 6, 7), had a higher CQ-binding ability than CS. CONCLUSION: The distribution and amount of each CQ-binding protein peak in both CS and CR strains of P. berghei were demonstrated, exhibiting differences in the ability of CQ-binding not only between CS and CR strains but also within the same strain. PMID- 12563772 TI - [Effect of artemether on nucleoside uptake and nucleic acid content in Schistosoma japonicum]. AB - AIM: To observe the effect of artemether (Art) on nucleoside uptake and nucleic acid content in Schistosoma japonicum. METHODS: RNA and DNA contents of both male and female worms harbored in mice treated intragastrically (i.g.) with Art 300 mg/kg for 24 h or 48 h were determined, respectively. After in vivo drug treatment, the schistosomes recovered were in vitro maintained in drug-free medium containing [3H]adenosine, [5-(3)H] uridine or [methyl-3H]thymidine at a final concentration of 37 MBq/L or 74 MBq/L for 2 h or 4 h, the tritiated nucleoside uptake and incorporation into nucleic acid of schistosomes were measured. RESULTS: The RNA and DNA contents of female worms recovered from the host 48 h after dosing were markedly decreased by 51.6% and 23.5%, respectively, while the RNA content of male worms showed 42.4% reduction. When the above mentioned schistosomes were in vitro exposed to the tritiated nucleoside for 2 h or 4 h, apparent decrease in tritiated nucleoside uptake with reduction rates of 35.2%-50.1% was seen in female worms. The incorporation of [methyl-3H]thymidine into the female worm DNA 2 h after incubation was reduced by 71.4% while the incorporation of [3H]adenosine into the female worm RNA and DNA 4 h after incubation was reduced by 65.2% and 50.0%, respectively. CONCLUSION: Art exhibited an apparent effect on the nucleic acid metabolism in schistosomes, especially in female worms. PMID- 12563773 TI - [A boy infant with Trichomonas vaginalis infection of the urethra]. PMID- 12563774 TI - [Evaluation of immunochromatographic test in the diagnosis of Plasmodium falciparum and Plasmodium vivax]. AB - AIM: To evaluate the effectiveness of immunochromatographic test(ICT) in detecting Plasmodium falciparum and Plasmodium vivax in malaria endemic areas. METHODS: ICT was used to detect P. falciparum and P. vivax among patients with fever in the outpatient clinics by comparason with thick blood smear method. RESULTS: The sensitivity of ICT to detect P. falciparum and P. vivax was 96.7% and 90.4%, respectively. The specificity of ICT was 98.6%, and the coincidence rate was 94.7%. There is no cross reaction between P. falciparum and P. vivax. CONCLUSION: ICT could detect P. falciparum and P. vivax simutaneously, being more rapid and simple than blood smear method. PMID- 12563775 TI - [Protective effects of leishmanial antigens against Leishmania infantum infection in Lagurus lagurus]. AB - AIM: To determine the protective effect of leishmanial surface antigens against experimental visceral leishmaniasis in Lagurus lagurus caused by Leishmania infantum. METHODS: Recombinant surface protein (rGP63) and lipophosphoglycan (LPG) of Leishmania were used with Corinebacterium parum vaccine as ajuvant to immunize Lagurus lagurus against a challenge with virulent strain of Leishmania infantum. The efficacy of immunoprotection was observed. RESULTS: When challenged with up to 2 x 10(7) promastigotes, the number of LD on the liver printing sections in the rGP63 + LPG + CP immunised animals was significantly decreased, the parasite reduction rate being 89.79%. LPG + CP gave a parasite reduction rate of 60.6% and rGP63/beta-galactosidase fusion protein + CP showed a parasite reduction rate of 42.45%. Purified rGP63 showed no protection. Immunization with rGP63 + LPG + CP followed by challenge inifection with 1 x 10(6), 5 x 10(6) and 1 x 10(7) promastigotes also showed significantly reduced infection rates. CONCLUSION: A combination of rGP63 + LPG + CP antigens could provide significant immunoprotection against L. infantum challenge in L. lagurus. PMID- 12563776 TI - [Field application of oral artesunate for preventing Schistosoma japonicum infection]. AB - AIM: To assess the preventive effect of oral artesunate against S. japonicum infection. METHODS: Residents in two pilots in the schistosomiasis endemic regions, 562 cases in Yanghe pilot, Wangjiang County and 218 cases in Shashan pilot, Guichi City, Anhui Province, were selected for this study. The residents were divided into two groups. Group I received artesunate 6 mg/kg once every 2 weeks for 4 times 2 wk after contacting with infested water from July to September in 1997. Group II received the same dosage of placebo at the corresponding times. Four weeks after the last administration, stool examination using hatching method and Kato's method was conducted to evaluate the effect. RESULTS: In Yanghe pilot, 2 cases were hatching positive in the artesunate treated group with an infection rate of 0.7% (2/273), while 11 cases were stool positive in placebo group with an infection rate of 3.8% (11/289) and a mean EPG of 26.40 +/- 1.49. In Shashan pilot, all cases in artesunate-treated group were stool negative, while 7 cases were stool positive in the placebo group with an infection rate of 6.3% (7/111), and a mean EPG of 14.23 +/- 2.14. The protection rate of artesunate was 80.9% and 100%, respectively, in the two pilots. CONCLUSION: Artesunate can protect the residents from S. japonicum infection effectively. PMID- 12563777 TI - [The role of chemotherapy in reducing the schistosom infection rate of livestock and schistosomiasis transmission in Dongting lake area]. PMID- 12563778 TI - [Mollusciciding action and toxicity of bromoacetamide]. PMID- 12563779 TI - [Progress in the research on expression system of recombinant malaria vaccine]. PMID- 12563780 TI - [Detection of heterophyid trematode egg in the blood of a case]. PMID- 12563781 TI - [Effect of water resource exploitation on endemicity of schistosomiasis]. PMID- 12563782 TI - [Purification and specific detection of two major Schistosoma gut-associated circulating antigens, CAA and CCA]. AB - AIM: To investigate the specificity differences of the 2 major gut-associated diagnostic circulating molecules in Schistosoma japonicum infection, CAA and CCA, and to obtain sufficient purified material for setting up a standard series in quantitative determinations. METHODS: Isolation and purification of the two worm fractions from a trichloroacetic acid (TCA) soluble preparation of S. japonicum adult worm antigen (AWAj-TCA) by Mono-Q anion exchange chromatography were performed and the specific reactivity of the eluted fractions by antigen-capture ELISA (specific for CAA or CCA) with reference to affinity purified preparations of S. mansoni CAA and CCA was analysed. RESULTS: By using an ionic strength gradient, CCA was eluted in two major peaks, an unbound fraction CCA-1, and a major bound fraction CCA-2. Two additional minor peaks, CCA-3 and CCA-4, were eluted at higher ionic strengths. CAA was only detected in a bound fraction, partly overlapping with CCA-3. In the CCA-1 and CCA-2 fractions reactivity was only found in the antigen-capture ELISA using anti-CCA McAbs both for capture and detection. The CAA fraction was predominantly found to be positive in the antigen capture ELISA using anti-CAA McAbs both for capture and detection. However, when using combinations of anti-CCA and anti-CAA McAbs for capture and detection by ELISA this fraction showed some reactivity. CONCLUSION: Two CCA fractions contain molecules which bear at least two CCA-epitopes; while the CAA fraction contains molecules which contain at least two CAA-epitopes, and one CCA-epitope. PMID- 12563783 TI - [Observation on the effect of Sj32DNA vaccine of Schistosoma japonicum]. AB - AIM: To explore the protective immunity against cercariae challenge in mice immunized with Sj32DNA vaccine of Schistosoma japonicum. METHODS: Primers were designed and the full length and part cDNA encoding 32 kDa(Sj32) amplified from pSj32 by PCR were cloned into pBK-CMV. Eukaryotic expression vector pBK-Sj32-1 containing full-length Sj32 cDNA and pBK-Sj32-2 containing fragment of Sj32 cDNA were constructed. After selection and identification, two plasmids were transfected into NIH 3T3 cells. The expression of Sj32 in transfected cells was confirmed by IFA. After identification, the two plasmids were prepared and purified on batches. Each BALB/c mouse was vaccinated at weeks 0, 3, and 5, by injection with 100 micrograms of purified Sj32DNA into quadriceps muscle. Mice immunized with constructs and the control mice were challenged four weeks after final DNA injection, and worms and the number of eggs in the liver tissue were counted at 8 weeks after challenge. RESULTS: Perfusion and egg counts showed significant differences in terms of worm reduction rate 38.6%-30.9% and egg reduction rate 55.7%-55.2% between the vaccinated and control mice. CONCLUSION: Sj32 DNA vaccine could result in protection against a subsequent challenge of S. japonicum in mice. PMID- 12563784 TI - [Studies on the features of protective immune response induced by recombinant Sjc26GST of Schistosoma japonicum]. AB - AIM: To compare the differences in the protective immune response induced by reSjc26GST between C57BL/6 and BALB/c mice. METHODS: Mice were immunized subcutaneously with reSjc26GST emulsified with Freund's adjuvant. The specific antibody isotypes elicited by reSjc26GST, in vitro lymphocyte proliferation and cytokine responses to reSjc26GST were examined. RESULTS: The level of anti reSjc26GST IgG was higher in immunized C57BL/6 mice than in BALB/c mice, and examination of the IgG subclasses demonstrated that reSjc26GST-immunized C57BL/6 mice resulted in predominantly IgG1 and IgG2a/IgG2b antibody responses, whereas in BALB/c mice, IgG1 antibody. Cytokine production assays revealed that the quantity of IFN-gamma and IL-2 and IL-5 released from the reSjc26GST-primed splenocytes were significantly higher in C57BL/6 mice than in BALB/c mice. These results showed that the immunization with reSjc26GST in C57BL/6 mice elicited a mixed immune reaction of Th1 and Th2 cell responses as demonstrated by the elevated levels of IgG1, IgG2a/IgG2b and IL-2, IFN-gamma, IL-5, whereas in BALB/c mice, reSjc26GST immunization elicited Th2 response as demonstrated by the elevated levels of IgG1, IL-2 and IFN-gamma. CONCLUSION: Immunization with reSjc26GST could elicit higher humoral and cellular immune responses in C57BL/6 mice than in BALB/c mice. PMID- 12563785 TI - [Evaluation of diagnostic value of 18 kDa antigen in alveolar echinococcosis]. AB - AIM: To evaluate the 18 kDa antigen in diagnosis of human alveolar echinococcosis. METHODS: A total of 214 sera from patients with alveolar echinococcosis (AE, 33), cystic echinococcosis (CE, 69), cysticercosis (30) and healthy controls (82) were examined by Western blotting to detect the antibodies against 18 kDa antigen using both antigens from protoscoleces of E. granulosus (Eg) and E. multilocularis (Em). All the sera were tested simultaneously by ELISA to detect the antibodies against hydatid cystic fluid antigens of sheep as a comparison. RESULTS: The positive rate by ELISA was 93.9% in AE, 85.5% in CE, 50% in cysticercosis and 6.1% in healthy controls, respectively, suggesting a critical cross-reaction and a certain degree of nonspecific reaction. The serum positive rates by Western blotting with the 18 kDa antigen of Eg and Em were 90.9% and 90.9% in AE, 10.1% and 13% in CE, 13.3% and 16.7% in cysticercosis, respectively. No positive was found in the sera from healthy controls. CONCLUSION: The 18 kDa antigen may be used in the differential diagnosis of AE from CE. PMID- 12563786 TI - [Sequencing of cytochrome C oxidase 1 gene of Ancylostoma duodenale and Necator americanus]. AB - AIM: To identify the genetic diversity between Ancylostoma duodenale and Necator americanus. METHODS: Mitochondrial cytochrome C oxidase subunit 1 (CO1) gene was amplified from genomic DNA of human hookworms collected from infected patients in Hejiang County, Sichuan Province, and the purified PCR products were directly sequenced by using Licor auto-sequencer. RESULTS: The PCR products were about 700 bp. Alignment of CO1 gene fragment sequences showed 89.7% similarity between Ancylostoma duodenale and Necator americanus, but still certain nucleotide variations (10.3%) existed. CONCLUSION: CO1 gene sequence can be used as a marker to identify the two species of human hookworms. PMID- 12563787 TI - [Studies on allozyme of Gammatricula]. AB - AIM: To furnish molecular genetic evidences for taxonomy of Gammatricula. METHODS: A total of 24 enzymes of 6 populations of Gammatricula songi and 1 population of Gammatricula chinensis collected from Kaihua County and Chunan County of Zhejiang Province were studied using horizontal starch gel electrophoresis. RESULTS: 29 loci were found. The percentages of polymorphic loci of G. songi populations were 6.9%-13.8%. All loci of G. chinensis were monomorphic. The Nei's distance among G. songi populations did not exceed 0.12. The Nei's distance between G. songi and G. chinensis was 0.73. CONCLUSION: The allozyme variations of inter-G. songi are limited, but the allozyme variation between G. songi and G. chinensis is significant. PMID- 12563788 TI - [Anti-hemolytic and anti-membranous lipid peroxidation effects of daphnetin]. AB - AIM: To investigate the anti-hemolytic and anti-membranous lipid peroxidation effects of daphnetin. METHODS: The inhibition rates of daphnetin on AQD-induced hemolysis and erythrocytic membranous lipid peroxidation were determined by routine in vitro assay for detecting hemolytic toxicity and by HPLC for detecting membranous lipid peroxidation products. RESULTS: Daphnetin at a range of 10-80 mumol/L inhibited dose-dependently AQD-induced hemolysis and lipid peroxidation by 33.0%-69.2% and 11.9%-58.2%, respectively. CONCLUSION: Daphnetin has anti hemolytic and anti-erythrocytic membranous lipid peroxidation effects. PMID- 12563789 TI - [Studies on recombinant chitinase and SXP-1 antigens as antimicrofilarial vaccines]. AB - AIM: To determine whether immunization with recombinant filarial chitinase or a fragment containing the epitope recognized by McAbMF1 and SXP-1 could protect jirds against microfilaremia resulting from infection with B. malayi. METHODS: Test jirds were immunized with the following recombinant parasite antigens: filarial chitinase, the c-terminal fragments F7R2 or F8R2 of r-chitinase, filarial SXP-1, myosin or maltose binding protein (MBP). Employing immunochemical techniqe (SDS-PAGE, Western Blotting) and serology (ELISA) measured antifilarial antibodies level. RESULTS: Immunization of jirds with recombinant chitinase induced partial protection against microfilaremia resulting from subsequent infection with B. malayi, but did not reduce adult worm burdens. Vaccination was much less effective when administered during the prepatent stage of infection and was ineffective when given to microfilaremic jirds. Immunization of jirds with SXP-1, an antigen present in multiple worm stages also reduced microfilaremia and, in some experiments, adult worm burdens. CONCLUSION: The recombinant chitinase, fragments F7R2 and F8R2 and SXP-1 could provide partial protection against microfilaremia in jirds. PMID- 12563790 TI - [First discovery of Taenia saginata asiatica infection in Yunnan province]. AB - AIM: To investigate the aetiology, species and epidemiological factors of Taenia infection in a pilot area of Lanping County, Yunnan Province. METHODS: Two patients with taeniasis were treated with antiscolex capsule and praziquantel, respectively. Five local weaning pigs free from tapeworm infection were fed with gravid proglottids obtained from the patients. After 2-3 months, the pigs were necropsied to examine cysticerci. In addition, one pig naturally infected with Taenia was treated as well. RESULTS: Based on morphological characteristics of the scolex and gravid proglottid of four tapeworms obtained from the patients, the worms were similar to T. saginata. 23 cysticerci recovered from two experimentally infected pigs were found in the livers, omentum majus and mesenterium, and 3 cysticerci were found in the liver and omentum majus in a naturally infected pig as well. The protoscolex of mature Cysticercus had two rows of rudimentary hooklets, one rostellum and four acetabula resembling to C. cellulosae. According to morphological characteristics and the location of cysticerci, the tapeworm was identified as Taenia saginata asiatia. CONCLUSION: T. saginata asiatica infection was first reported in Yunnan Province. PMID- 12563791 TI - [Association expression of genes encoding gst of Schistosoma japonicum and enterotoxigenic Escherichia coli]. AB - AIM: To study the association expression of two genes encoding GST of Schistosoma japonicum and K99 of enterotoxigenic Escherichia coli. METHODS: PCR technique was used to gain the K99 gene. After digestion with BamH I and EcoR I, the gene was cloned into the plasmid vector pUC18 by using recombinant DNA techniques. The other target gene of GST in pSj5 plasmid was obtained by EcoR I digestion, and was then ligated into the recombinant plasmid pUC18-K99. The expressed product was assayed by SDS-PAGE, Western blotting, reversal IHA, ELISA and transmission electron microscopy. RESULTS: Co-expression product of around 50 kDa was obtained. The protein was recognized by anti-GST and anti-K99 antibodies (ELISA and IHA), and acted as pilus on the surface of transformed DH5 alpha E. coli bacteria. CONCLUSION: Co-expression of the genes encoding GST and K99 was successfully achieved. PMID- 12563792 TI - [Studies on immunoprotection of monoclonal antibodies against Cryptosporidium parvum]. AB - AIM: To explore the immune protection of the monoclonal antibody (McAb) against Cryptosporidium parvum. METHODS: On the basis of Madin-Darby canine kidney (MDCK) cell culture, the immunoprotection of McAb against C. parvum was screened by means of neutralization test in vitro, and verified by rat model and transmission electron microscopy of infected MDCK cells. RESULTS: The number of C. parvum sporozoites recovered on the surface of rat intestinal epithelial cells and the mean output of oocyst were significantly reduced by McAb Z3D2 (P < 0.001, P < 0.05). The decreased Cryptos poridium at each development phase within infected MDCK cells and less damage of cell ultrastructure caused by Cryptosporidium were found in Z3D2 treated group. CONCLUSION: McAb Z3D2 possesses highly effective protection against C. parvum. PMID- 12563793 TI - [Effect of IL-6 on the multiplication of Toxoplasma gondii]. AB - AIM: To observe the effect of IL-6 on the multiplication of Toxoplasma gondii. METHODS: Peritoneal macrophages from C57BL/6 mouse were incubated with 3H-uracil labelled T. gondii in vitro. RESULTS: Pretreatment (but not post-treatment) of Mphi with IL-6 enhanced T. gondii multiplication in a dose-dependent manner. Pretreatment with IFN gamma resulted in active killing of parasites whereas the addition of IL-6 resulted in a partial reversal of IFN gamma-mediated toxoplasmacidal activity. Combining TNF alpha with IL-6 and IFN gamma pretreatment resulted in restoration of toxoplasmacidal activity. Addition of polyclonal anti-TNF alpha antibodies to IL-6 and IFN gamma pretreatment resulted in enhancement in the IL-6-mediated impairment of IFN gamma function. CONCLUSION: IL-6 could enhance intraperitoneal multiplication of T. gondii and reverse IFN gamma-mediated toxoplasmacidal activity. PMID- 12563794 TI - [Establishment of a multiplex PCR system to detect plasmodium]. AB - AIM: To establish a simple, rapid and practical multiplex PCR system to detect Plasmodium vivax(P. v) and Plasmodium falciparum(P. f). METHODS: A common upper primer S1 and two species-specific lower primers of P. v and P. f, S2, S3 were designed according to the sequences of the small subunit ribosomal DNA (SSUrDNA) fragments of the two Plasmodium species. Using these three oligonucleotide primers, the two-temperature-point multiplex PCR system was established and applied to detect P. v and P. f in the stock blood samples of clinically confirmed patients. RESULTS: DNA fragments of about 705 and 575 base pairs were successfully amplified by multiplex PCR from the genomic DNA of P. v and P. f, but no fragments were obtained from that of P. knowlesi, P. cynomolgi and blood of healthy persons. By means of restriction endonuclease digestion, the amplified fragments were confirmed to be the SSUrDNA fragments of P. v and P. f as expected. This method was successfully used in detecting parasitemia 2-10 parasite/microliter whole blood. Of 104 samples tested by this system, 81 were coincident with microscopic examination. The multiplex PCR system also found 17 samples of mixed infection, which were not detected microscopically. Another 2 samples detected as P. v by microscopic examination were verified to be P. f infection by the multiplex PCR. CONCLUSION: The ease of operation together with high sensitivity and specificity, particularly the sensitive detection for mixed infection in a single run of amplification suggests that the multiplex PCR system might be a useful tool for malaria diagnosis. PMID- 12563795 TI - [Preparation of DNA probe for Cryptosporidium parvum]. AB - AIM: To prepare a probe with high specificity and sensitivity for the detection of Cryptosporidium parvum. METHODS: Using PCR method, a fragment from the DNA of C. parvum was amplified. The PCR product, 452 bp DNA, was labeled with hapten digoxigenin. RESULTS: Examination of sensitivity showed that the DNA probe could detect as low as 2 pg DNA from C. parvum. The dot-blot hybridizition assay showed that the probe hybridized with the DNA of C. parvum, but not hybridized with DNA of E. histolytica, G. lamblia, E. coli and D. bacilli. CONCLUSION: The probe was highly specific and sensitive for the detection of C. parvum. PMID- 12563796 TI - [Preliminary studies on microwave irradiation ELISA for diagnosis of Schistosomiasis japonica]. AB - AIM: To explore a fast and highly efficient method for the diagnosis of schistosomiasis japonica. METHODS: Using microwave irradiation ELISA (MWI-ELISA) and fast-ELISA to detect specific antibodies in sera from 118 cases with schistosomiasis japonica, 61 healthy individuals and 12 paragonimiasis cases. RESULTS: The positive rates of schistosomiasis cases were 88.1% (104/118) by MWI ELISA and 91.5% (108/118) by fast-ELISA, respectively (chi 2 = 0.74, P > 0.05). The false positive reaction of healthy individuals was found in 2 cases(3.3%) by MWI-ELISA and 1 case (1.6%) by fast-ELISA, respectively (chi 2 = 0.34, P > 0.05). No false positive reaction was found in paragonimiasis cases. CONCLUSION: The sensitivity and specificity of the two tests were similar, however, MWI-ELISA was faster than fast-ELISA. PMID- 12563797 TI - [Detecton of circulating antigen in sera from mice infected with Toxoplasma tachyzoites]. AB - AIM: To explore an assay method for the early diagnosis of Toxoplasma infection. METHODS: Serum samples collected from three groups of mice infected experimentally with different doses of Toxoplasma trophozoites were detected for the presence of Toxoplasma circulating antigen by using fast ELISA. RESULTS: Toxoplasma circulating antigen was detected on days 4, 4 and 3 after infection in light, moderate and heavy infection groups, respectively. CONCLUSION: The level of circulating antigen was in parallel with the duration of infection. The determination of circulating antigen is useful in the early diagnosis of Toxoplasma infection. PMID- 12563798 TI - [Advances in researches on anti-Cryptosporidium parvum hyperimmune bovine colostrum]. PMID- 12563799 TI - [Susceptibility of Oncomelania snail from MiaoRiver area in Hubei province to Schistosoma japonica]. PMID- 12563800 TI - [Long-term effect of health education in schistosome infection control among school children in a lake region]. PMID- 12563801 TI - [Diagnosis of cerebral cysticercosis in children]. PMID- 12563802 TI - [Controlling schistosomiasis transmission by combining with agricultural development in lake regions]. PMID- 12563803 TI - [Investigation of Enterobius infection in children in the rural area of Ji'nan]. PMID- 12563804 TI - [Experimental observation on killing of maggots with Macleay codata]. PMID- 12563805 TI - [A pilot study on malaria control by using a new strategy of combining strengthening infection source treatment and health education in mountainous areas of Hainan province]. AB - AIM: To explore a new malaria control strategy that fits current epiodemiological condition and coincides with modern medicine model and the principle of cost benefit. METHODS: The new strategy highlights the risk villages and risk population as the focal point and integrates health education with behavioral intervention. The main anti-malaria measures consists of carrying out health education in risk villages, giving mass drug administrations in risk population staying overnight in the mountain, following up malaria cases for implementing radical cure, but without using traditional residual spraying or impregnating bednets with insecticides. RESULTS: After having adopted the new strategy and taken the control measures, the people's knowledge about malaria increased to a higher level and the indices of malaria reduced to a lower level. The rate of bednet-using in the population was increased from 26.8% to 72.6%. The annual parasite incidence (API) of malaria was declined from 3.5% in 1994 to 1.1% in 1996 and 0.8% in 1997, and the API of falciparum malaria was declined from 1.0% to 0.3% and 0.3% respectively in the townships at the same time. The parasite rate(PR) of malaria was declined from 7.2% in May, 1995 to 2.1% in November, 1996 and 1.2% in October, 1997 and the PR of falciparum malaria was declined from 1.2% in May, 1995 to 0.1% in October, 1997. The proportion of villages without malaria cases was increased from 18.6% in 1994 to 54.2% in 1997, and the number of risk villages with a malaria incidence above 5% was reduced from 14 to 2 at the same time. The ratio of cost/benefit was 1:2.4 in 1995-1996 and 1:4.4 in 1997, showing a better economic benefit. CONCLUSION: The expectant result has been obtained, thereby providing new experience for the malaria control in the mountainous areas of Hainan Province. PMID- 12563806 TI - [Determination of circulating antigen in urine of rabbits infected with Schistosoma japonicum]. AB - AIM: To develop an immunoassay for the detection of circulating schistosome antigen (CSA) in host urine. METHODS: CSA extracted from the urine of the rabbits infected with Schistosoma japonicum was used for preparing and selecting specific monoclonal antibody. A one-step dot-ELISA using this specific McAb and a 2nd antibody was used for detection of circulating schistosome antigen in the urine of infected rabbits. RESULTS: No CSA was detected in urine from all of 22 rabbits before infection. The positive rate of CSA in infected rabbits was correlated with the intensity of infection and the time of infection. No CSA was detected in the urine rabbits of 3 weeks after infection with 25 cercariae while the positive rate of CSA in the urine from rabbits infected with 200 cercariae was 40% and 100% after 3 and 6 weeks, respectively. A combination of CSA detection in both urine and serum may increase the detectability. CONCLUSION: The specific McAb prepared by urine CSA of Schistosoma japonicum-infected rabbits can be used as a probe for detecting CSA in the urine of infected rabbits. PMID- 12563807 TI - [In vivo anti-fecundity effect of recombinant Sjc26 GST against Schistosoma japonicum in immunized mice]. AB - AIM: To observe the anti-fecundity effect of recombinant Sjc26 GST against Schistosoma japonicum and its mechanism. METHODS: Male ICR mice were immunized with rSjc26 GST. Five days after final injection, mice of the immunization and control groups were challenged with 40 +/- 1 S. japonicum cercariae. All mice were sacrified at 6 weeks after challenge. RESULTS: The worm reduction rate was 30.0%, and the egg reduction rates in the liver and in the spleen were 57.1% and 79.9%, respectively. Inhibition of the vitelline gland and testis was also observed by transmission electron microscopy. The number of vitelline globes, vitelline droplets and lipid droplets in the cytoplasm of vitelline cell and the number of lipid droplets in the supporting cells and the number of spermatids in the testis were apparently reduced. CONCLUSION: The damage of the main reproductive organs of Schistosoma japonicum was one of the factors responsible for rSjc26 GST-bound anti-fecundity action. PMID- 12563808 TI - [Recombination and cloning of MSP1(19) and PfCMR of Plasmodium falciparum]. AB - AIM: To construct a recombinant plasmid DNA encoding multiantigens of Plasmodium falciparum and to provide the requirements for DNA immunization. METHODS: Two oligonucleotide primers were designed to amplify MSP1(19), the purified PCR products were digested by Sal I + Xba I, and the recombinant plasmid pWR450 1/PfCMR was digested by EcoR I + Sal I to recover PfCMR gene. PfCMR and MSP1(19) gene fragments were linked and recombined with mammalian expression vector pcDNA3. RESULTS: The MSP1(19) gene fragment with about 363 base pairs were specifically amplified by using PCR technique. The positive recombinant pcDNA3 PfCMR-MSP1(19) (named pcDNA3-Pf8) was screened and identified by agarose gel electrophoresis, endonulease digestion and PCR technique, the whole length of Pf8 is 618 bp. CONCLUSION: By specifically amplifying MSP1(19) gene at the C-terminal of MSP1, a recombinant plasmid pcDNA3-Pf8 encoding multiantigens of Plasmodium falciparum was successfully constructed. PMID- 12563809 TI - [Differences in haemozoin production and pathogenicity between chloroquine sensitive and chloroquine-resistant strains of Plasmodium berghei]. AB - AIM: To better understand the differences in haemozoin formation and pathogenicity between chloroquine-sensitive(N) and chloroquine-resistant(RC) strains of Plasmodium berghei. METHODS: IRC mice were grouped as follows: group Xba I (Normal control, NC), group II (control treated with chloroquine alone, CC), group III (mice infected with N strain), group IV (mice infected with RC strain, RC) and group V (mice infected with RC strain and treated with chloroquine, RCC). Morphologic features of the parasites, parasitaemia, histological and ultrastructural changes of livers among the groups were compared. RESULTS: Severe damages of the hepatic cells of the N group including increased lysosomes and swollen and fused mitochondria were detected. On the contrary, the prominent features in liver section of the RC group were inflammatory cell (notably mononuclear) infiltration and Kupffer cell activation. Numerous trophozoites and schizonts were sequestrated in hepatic sinusoids and less degeneration of parenchymal cells was found except for some swollen and vacuolated mitochondria. Internal food vacuoles containing haemozoin were found in the parasites of the N group, whereas external food vacuoles without haemozoin granules were arranged in foamy appearance within the parasitized cell of the RC group. CONCLUSION: P. berghei RC strain may modify the mode of ingestion and degradation of hemoglobin in the parasites, resulting in impeding haemozoin formation. The difference in virulence between the N and RC strain of P. berghei is probably attributed to the significant differences in the induction of immune response of the host. PMID- 12563810 TI - Testosterone inhibits apoptosis of Leishmania donovani-infected macrophages. AB - AIM: Investigate the effect of the male sex hormone, testosterone (Te), on apoptosis of bone marrow derived macrophages (BMMs) from female C57BL/6j mice METHODS: Propidium iodide (PI) staining and transmission electron microscopy (TEM) techniques were used to investigate apoptosis specific-morphological changes. BMMs derived from Te-treated mice and Oil-treated mice were challenged with Leishmania donovani (L. d.), Oligo-nucleosomal DNA were extracted 24 h post infection to detect apoptosis. RESULTS: The removal of M-CSF from the medium could induce BMM apoptofsis. And the DNA fragmentation assay also indicated that: 1. there was no difference in the amount of apoptotic cells between Te and Oil group; 2. Te + L. d. group had significantly less dead cells than Oil + L. d. group demonstrating that Te could prevent apoptosis of macrophage infected with L. d. to a greater extent. CONCLUSION: Te inhibits apoptosis of the macrophages infected with L. d., however, this inhibition did not occur in the macrophages uninfected with L. d. Te-induced macrophage apoptosis-inhibition may play an important role in Te-induced immunosuppression. PMID- 12563811 TI - [Observation on the clinical symptoms and sporocyst excretion in human volunteers experimentally infected with Sarcocystis hominis]. AB - AIM: To investigate the excretion of sporocysts and clinical manifestations in humans experimentally infected of Sarcocystis hominis. METHODS: Three volunteers were infected by eating raw beef containing cysts of S. hominis. One ingested about 1,567 cysts in skeletal muscles of a naturally infected cattle; two volunteers each ingested about 14,740 cysts from an experimentally infected water buffalo meat. Fecal examination by zinc sulfate flotation method was conducted daily since d4 postinfection (pi). RESULTS: Free sporocysts and oocysts were found in their faeces from d11-40, d12-23, d10-30 pi, and peaked at d18, d14, d14. All of them presented clinical symptoms such as abdominal pain, distension, watery diarrhea and eosinophilia 1 wk approximately 4 wk pi and were spontaneously cured within 29 days pi without taking any medicine. CONCLUSION: All the experimentally infected persons had gastrointestinal symptoms and passed sporocysts and oocysts in faeces 10-12 days after infection and persisted for 11 29 days. PMID- 12563812 TI - Diagnosis of trichinosis by indirect fluorescent antibody test with Trichinella larva section. AB - AIM: To evaluate the value of indirect fluorescent antibody test (IFAT) in diagnosing trichinosis. METHODS: IFAT using frozen section of the purified Trichinella larvae as antigen was employed to detect anti-Trichinella antibodies in sera from mice infected with Trichinella spiralis and the patients with trichinosis. Sera from patients with trichinosis were collected at different time intervals after onset of the disease and after treatment with albendazole. Sera from patients with other parasitic diseases and healthy individuals were used as control. RESULTS: The anti-Trichinella antibodies were detected in 15 infected mice as early as 2 weeks after infection, and in 86.5% of 467 patients with trichinosis. Sera from patients with filariasis, echinococcosis and healthy controls were all negative. 8 of 48 (16.7%) cases with paragonimiasis showed positive cross reaction. The antibody positive rate of patients with trichinosis was 70.2% one week after onset of disease and increased to 91.0%, 95.8% and 100% at two, three and four weeks after onset, respectively. The frozen section of the larvae can be stored at -20 degrees C for 5 years without loss of antigenicity. The IFAT positive rate was increased from 87.5% before treatment to 100% one week after treatment, and the serum antibody titers were elevated as well. The antibody negative conversion rate of the patients was 24% at one month after teratment and increased to 75% at four months after treatment. CONCLUSION: IFAT using frozen section of the purified Trichinella larvae as antigen is highly sensitive and specific for the diagnosis of trichinosis and evaluation of the therapeutic effect. PMID- 12563813 TI - [Changes in chloroquine resistance of Plasmodium falciparum in Hainan province]. AB - AIM: To observe the variation in resistance of Plasmodium falciparum after the cessation of chloroquine medication. METHODS: WHO standard in vitro microtest and in vivo test were used. RESULTS: In vitro test showed that the rate of chloroquine-resistant P. falciparum dropped from 97.9% in 1981 to 26.7% in 1997 (P < 0.001). The mean concentration of chloroquine for complete inhibition of schizont formation declined from 10.46 +/- 7.14 pmol/ul blood in 1981 to 1.63 +/- 1.47 pmol/microliter blood in 1997(P < 0.001). In vivo test showed that the rate of chloroquine-resistant P. falciparum decreased from 84.2% in 1981 to 18.4% in 1997(P < 0.001). The proportion of R III cases to the total resistant cases dropped from 53.1% in 1981 to 14.3% in 1997. CONCLUSION: A tendency of progressive decline of resistance of the parasite was revealed after the cessation of chloroquine medication. PMID- 12563814 TI - [Experimental therapy of Pagumogonimus skrjabini infection in rats with triclabendazole]. AB - AIM: To observe the therapeutic effect of triclabendazole in rats infected with Pagumogonimus skrjabini. METHODS: Metacercariae of P. skrjabini were isolated from crabs (Sinopotamon) collected from two endemic areas (Weiyuan County of Sichuan Province and Wansheng District of Chongqing Municipality). Wistar rats were each infected intraperitoneally with metacercariae. One month and two months after infection, the infected rats were treated with triclabendazole at the total dosage of 300 mg/kg over 2 days, 450 mg/kg over 3 days and 600 mg/kg over 3 days, respectively. RESULTS: The worm reduction rates were 50%, 80% and 87% respectively one month after completion of tretment. Dead worms (about 1 mm size) recovered from the muscles, liver, abdominal cavity, chest cavity and lungs were signicantly diminished in size and weight in comparison with those of the control group. Many large (about 1 cm) black-colored distended worm cysts usually with two adult worms and many eggs were found in the lungs of the control rats. Most worm cysts in the treated groups of rats were found to be atrophied, changing into hemorrhagic-necrotic patches. CONCLUSION: Triclabendazole is highly active against P. skrjabini in rats. PMID- 12563815 TI - [Molecular cloning of a cDNA encoding an antigenic polypeptide containing repeat units of Spirometra erinaceieuropaei plerocercoid]. AB - AIM: To study the gene structure encoding some antigenic polypeptides of plerocercoid of Spirometra erinaceieuropaei (SEP). METHODS: A cDNA library constructed from SEP was immunoscreened using mouse anti-SEP polyclonal antibody. The gene structure was analyzed by computer after the insert of positive clone was subcloned and the nucleotide sequences of the insert were determined. The length of mRNA hybridized with cDNA was detected by Northern blotting. RESULTS: A cDNA clone of 1,084 bp encoding an antigenic polypeptide was isolated. The clone contained one open reading frame composed of 828 bp encoding 276 amino acids. The open reading frame contained tandem repeating unit of 123 bp which appeared 5 times in this clone. The 41 amino acids deduced from 123 bp repeating unit consisted of 53.7% of hydrophobic amino acid residues. In Northern blot assay of poly (A)+RNA, a strong band of about 1.1 kb and a weak band of about 1.7 kb were found in the plerocercoids but no band was found in the adult worms. CONCLUSION: The repeating element derived from the plerocercoids may be associated with the migration of the plerocercoids in the host tissue. The polypeptides with repeating element may act as evasive antigens of the parasite to escape from the destruction of host immune reactions. PMID- 12563816 TI - [Assay of sensitivity of Plasmodium falciparum to chloroquine, amodiaquine, piperaquine, mefloquine and quinine in Yunnan province]. AB - AIM: To determine the sensitivity of P. falciparum to chloroquine, amodiaquine, piperaquine, mefloquine and quinine in Yunnan Province of China in 1992-1995. METHODS: Rieckmann's in vitro microtechnigue was used. The sensitivity of P. falciparum was tested to the above-mentioned antimalarials. RESULTS: The resistance rates of isolates of P. falciparum from the south, southeast and the west part of Yunnan to chloroquine and amodiaquine were 96.7% (29/30), 78.9% (30/38), 95.7% (22/23) and 100% (30/30), 85.3% (29/34), 8/9, respectively, with their corresponding ID50 of 125 nmol/L, 136 nmol/L and 176 nmol/L, and 52 nmol/L, 64 nmol/L and 72 nmol/L, respectively. All the isolates were sensitive to quinine and their ID50 were 480 nmol/L, 352 nmol/L, 608 nmol/L, respectively. The resistance rates of P. falciparum from the south part and southeast part of Yunnan to piperaquine were 96.4% (27/29), 72.9 (27/37), respectively, their ID50 were 320 nmol/L and 228 nmol/L; all the cases were sensitive to mefloqine, their ID50 were 68 nmol/L and 88 nmol/L. CONCLUSION: P. falciparum generally produces resistance to chloroquine, amodiaquine and piperaquine in Yunnan Province; the degree of resistance to chloroquine of P. falciparum from the west part of Yunnan were higher than the P. falciparum from the southeast part of Yunnan; all the isolates were sensitive to mefloquine and quinine in this region. PMID- 12563817 TI - [Advances in molecular biological research on cytochrome P450 of insects]. PMID- 12563818 TI - [Molecular mechanism of virulence variation of Toxoplasma strains]. PMID- 12563819 TI - [Effect of schistosomiasis on the growth, physical fitness and working capacity of the human population]. PMID- 12563820 TI - [Data analysis of human trichinosis epidemic in Yunnan province from 1964 to 1997]. PMID- 12563821 TI - [Studies on the degradation of Schistosoma japonicum proteinase on hemoglobins of different hosts]. PMID- 12563822 TI - [Clinical analysis of 30 cases of toxoplasmosis]. PMID- 12563823 TI - [Clinical special manifestations of 4 cases with vivax malaria]. PMID- 12563824 TI - [Preliminary observation on detection of Schistosoma japonicum cercariae in the water of marshland]. PMID- 12563825 TI - [Three cases of cysticercosis misdiagnosed as brain tumor]. PMID- 12563826 TI - [Establishment and application of dipstick sandwich colloidal dye immunoassay for circulating antigen detection in schistosomiasis patients]. AB - AIM: To set up a simple immunoassay for the detection of circulating antigens in host serum. METHODS: A kind of colloidal pale purple dye (HFRL) used as a staining reagent for immunoassay was first selected from the dyes produced in China and an optimum condition for labelling the dye onto the combined IgM antibodies was explored. A dipstick sandwich colloidal dye immunoassay (DS-DIA) for the detection of circulating antigens in host sera was established. RESULTS: The minimal concentration of SEA detected was 5 ng/ml by DS-DIA. Serum circulating antigen detection in 14 cases of acute schistosomiasis japonica, 113 chronic cases and in 113 healthy controls revealed that the sensitivity was 100% in acute cases, 52.2% in chronic cases and the specificity was 92.9% for uninfected students. The sensitivity and specificity of DIA were similar to those of dot-ELISA. A certain degree of diagnostic complementarity up to 76.1% in sera from these patients was seen when DS-DIA and dot-ELISA were used in paralle. CONCLUSION: DS-DIA is a simple, economical and reliable method for detecting circulating antigens of Schistosoma japonicum, having a wide potential value especially for field use. PMID- 12563827 TI - [Preliminary study on the levels of natural antibodies against Schistosoma japonicum in Microtus fortis in Dongting lake area]. AB - AIM: To explore the detection rate and the levels of natural antibodies to cercarial antigen(CA), juvenile antigen(JA), adult worm antigen (AWA) and soluble egg antigen (SEA), respectively, derived from Schistosoma japoncum in Microtus fortis distributed in Dongting Lake area. METHODS: The antibodies in wild Microtus fortis(WMF), laboratory-bred Microtus fortis (BMF) and laboratory-bred Microtus fortis 15 days after S. japonicum infection (BMFi15) were detected by indirect ELISA and a comparison of the antibody levels between Microtus fortis including WMF, BMF, BMFi15 and albino mice without infection (AM), 15 days after infection(AMi15) and 42 days after infection (AMi42) was made. RESULTS: The positive rates of detection for anti-JA-Ab in WMF and BMF were 94.6% and 94.4%, respectively, followed by 85.5% and 83.3% for anti-AWA-Ab, 58.1% and 59.4% for anti-SEA-Ab, and 14.3% and 13.9% for anti-CA-Ab. The levels of antibodies to the above four antigens in BMFi15 increased significantly, the detection rates being 100% for anti-JA-Ab and anti-AWA-Ab, 84.4% for anti-SEA-Ab and 65.6% for anti-CA Ab. The levels of antibodies in WME, BMF and BMFi15 had the similar trend(JA > AWA > SEA > CA), and those in BMFi15 to CA, JA, AWA and SEA increased by 1.9, 2.2, 2.1 and 1.5 folds, respectively, when compared with BMF. CONCLUSION: Natural antibodies to Schistosoma japonicum were found in wild and labortorybred Microtus fortis. The levels of antibodies in BMFi15 to four above antigens were apparently higher than those in AMi15. PMID- 12563828 TI - [Taxonomic studies on Echinochasmus fujienensis and its related species by random amplified polymorphic DNA analysis and experimental infection]. AB - AIM: To explore the identification and differentiation of Echinochasmus fujianensis, Echinochasmus japonicus(Jiangxi strain and Fujian strain), Echinochasmus liliputanus Anhui and Echinochasmus per foliatus Hubei. METHODS: Random amplified polymorphic DNA analysis (RAPD) and experimental animal infection were performed. RESULTS: 469 polymorphic DNA fragments were obtained by 28 primers from 4 Echinochasmus species and strains in Fujian, Anhui and Jiangxi. 20.8% and 97.6% of the fragments in Echinochasmus fujianensis were the same as those in Echinochasmus japonicus Fujian strain and in Echinochasmus liliputanus Anhui, respectively, 99.7% of the fragments were the same between Echinochasmus japonicus Jiangxi strain and Fujian strain. CONCLUSION: Echinochasmus fujianensis and Echinochasmus liliputanus Anhui are the same species. Echinochasmus fujienensis is an independent species different from Echinochasmus japonicus. Polyinfection of Echinochasmus fujianensis, Echinochasmus japonicus and Echinochasmus perfoliatus exist in all the 3 provinces, Hubei, Anbui and Fujian, of which Echinochasmus fujienensis is a dominant species. PMID- 12563829 TI - [Changes in transforming growth factor beta 1 mRNA and its clinical value in patients with schistosomal liver fibrosis]. AB - AIM: To explore the changes in TGF-beta 1 mRNA in patients with schistosomal liver fibrosis. METHODS: Reverse-transcription polymerase chain reaction and dot blot analysis were used to determine the level of TGF-beta 1 mRNA in PBMC. The serum levels of hyaluronic acid, type IV collagen, laminine and procollagen III peptide were also measured simultaneously. RESULTS: The mean level (m +/- s) of PBMC TGF-beta 1 mRNA was significantly higher in late-stage schistosomiasis group (LSS) (1.26 +/- 0.14), liver cirrhosis group (LC) (2.05 +/- 0.81) and hepatocellular carcinoma group (HCC) than that in control group (0.62 +/- 0.40) (P < 0.05). PBMC TGF-beta 1 mRNA level was higher in LC and HCC than that in LSS (P < 0.05). No significant difference was found in TGF-beta 1 mRNA between PBMC and liver tissue in patients with HCC. All patients with increased fibrogenic activity (i.e., abnormal levels of serum HA, Col-IV and LN) had increased levels of TGF-beta 1 mRNA. CONCLUSION: The level of TGF-beta 1 mRNA is correlated with the fibrotic activity of liver, the increasement of TGF-beta 1 mRNA in schistosomiasis patients after praziquantel treatment might induce liver fibrosis. PMID- 12563830 TI - [Cloning and sequence analysis of histidine-rich protein-II gene fragment of Plasmodium falciparum yunnan strain]. AB - AIM: To determine the nucleotide sequence of the partial gene of the histidine rich protein II (HRP II) the Plasmodium falciparum PFD-3/YN and find out the differences of the HRP II gene sequence between this isolate and other isolates. METHODS: The histidine-rich protein-II gene fragment was amplified by polymerase chain reaction and cloned into M13 bacteriophage. M13-HRP II single strand DNAs of three positive clones were extracted, respectively. The nucleotide sequence of the HRP II gene fragment was determined by the dideoxy chain termination method. Using PCGENE software to compare and analyze the HRP II gene sequence among the different isolates. RESULTS: Different degrees of diversity of the HRP II gene sequences were found among Plasmodium falciparum PFD-3/YN(from China) and two other isolates (7G8 from Brazil and D10 from Gambia). The HRP II in the three isolates exhibited 70.3% homology in amino acid sequences and 68.6% homology in the nucleic acid sequences. CONCLUSION: There were differences in HRP II gene sequence among the Plasmodium falciparum PFD-3/YN and two other isolates (7G8 and D10). PMID- 12563831 TI - [Detection of circulating antigen in serum and cerebrospinal fluid for diagnosis of cerebral cysticercosis]. AB - AIM: To detect circulating antigen(CAg) in serum and cerebrospinal fluid(CSF) from patients with cerebral cysticercosis with specific antiserum by sandwich ELISA. METHODS: Antisera were prepared from rabbits immunized respectively with 3 antigens with molecule weights of 64 kDa, 53 kDa, 32 kDa-30 kDa extracted from Cysticercus cellulosae and purified by SDS-PAGE. RESULTS: When samples from patients were tested with antiserum against 53 kDa antigen, the CAg positive rate was 93.8% in serum and 91.7% in CSF of 32 patients with active cysticercosis, whereas only one positive was found in CSF in 16 patients with inactive cysticercosis. The detection rate of CAg was significantly higher with anti-53 kDa antiserum than with anti-64 kDa and anti-32 kDa-30 kDa antiserum. CONCLUSION: Sandwich-ELISA using antiserum against 53 kDa antigen to detect CAg was found to be a promising assay for diagnosis and evaluation of treatment efficacy of active cerebral cysticercosis in terms of its high sensitivity and specificity. PMID- 12563832 TI - [Study on family aggregation of cases of advanced schistosomiasis japonica]. AB - AIM: To explore the family aggregation of advanced schistosomiasis japonica. METHODS: Eighty-one cases of advanced schistosomiasis(AS) and 67 cases of non advanced schistosomiasis with history of infections in Yushan County, Jiangxi Province were chosen as proband groups and control groups respectively, then grades 1 and 2 relatives of them were investigated on AS. Family aggregation of AS was analyzed through comparing the prevalence rate between the close and distant relatives of probands and controls and fitting the observed distribution of AS cases among the population by zero-truncated Poisson distribution and zero truncated negative binomial distribution. RESULTS: The prevalence rate was higher in the close relatives (Group I relatives) of the probands than in the distant relatives(Group II relatives) of the probands and in the controls' relatives. The observed distribution of AS was beyond the probability of the zero-truncated Poisson distribution, but consistent with the zero-truncated negative binomial distribution. CONCLUSION: Family aggregation of advanced schistosomiasis does exist. PMID- 12563833 TI - [Morpholgical, histological and histochemical observations on the effect of albendazole on encysted larvae of Trichinella spiralis in mice]. AB - AIM: To study the mechanism of action of albendazole on encysted larvae of Trichinella spiralis in mice. METHODS: Twelve Kunming strain mice each infected with 200 T. spiralis larvae were equally divided into two groups. Six mice of the treatment group were treated with 30 mg of albendazole/kg daily for 5 days. Morphological, histological and histochemical methods were used. RESULTS: The encysted larvae of albendazole-treated group became significantly damaged, most worms were shrunken and surrounded by inflammatory cells. Histochemical study demonstrated that the glycogen and RNA content of the larvae in the treated group was decreased, the activities of SDH, ATPase, ACP were lower than those of the control group. CONCLUSION: Albendazole can affect the physiological function of Trichinella spiralis. PMID- 12563834 TI - [Genotype and sequence analysis of merozoite surface protein 1 of Plasmodium falciparum isolates in Yunnan province]. AB - AIM: To identify the genotype of merozoite surface protein 1 of Plasmodium falciparum in Yunnan Province and explore the polymorphism of MSP1 genes in geographical characteristics and genetics. METHODS: Nested polymerase chain reaction was applied to genotyping of P. falciparum isolated in Yunnan Province. Some of parasite alleles were sequenced by dye primer cycle sequencing. RESULTS: In 30 P. falciparum infections, 38 different alleles were found. Of them, the dominant allele was a variant of MAD20, while was K1 less and the RO33 was few. In addition, incidences of mixed allele infections were observed. Sequence analysis showed that DNA sequences of MAD20, K1 and RO33 alleles from Yunnan were highly homologous with those of international standard strains, respectively. CONCLUSION: Composition and sequence characteristics of P. falciparum arasite population in the endemic area can be detected by genotyping with MSP1 as genetic marker, which would be useful for the prevention and treatment of malaria. PMID- 12563835 TI - [Preparation of monoclonal antibodies by intrasplenic immunization of mice with urine circulating antigen from patients infected with Schistosoma japonicum]. AB - AIM: To explore an efficient method for preparing monoclonal antibodies against circulating antigen of S. japonicum in the urine of schistosomiasis patients. METHODS: The urine of schistosomasis patients were precipitated with 5% trichloroacetic acid, and then used to immunize the BALB/c mice by intrasplenic immunization to prepare McAb. RESULTS: Two monoclonal cell lines against the CAg in the urine of schistosomiasis patients have been established. The immunoglobulin subclass of the 2 McAbs (2E6 and 2B11) were identified as IgM. The titres of 2E6 and 2B11 were 2.56 x 10(5) and 6.4 x 10(4). On immunodiffusion, McAb 2E6 showed one precipitation line with IHU-CAg, however, both McAbs did not react with NHU-CAg. Using ascitic fluid of 2B11 and 2E6 as capture antibody separately, and HRP-H11 as labelled antibody to detect the urine of 12 acute schistosomasis patients, 2B11 2E6 gave 6 and 3 positive, respectively, whereas 8 normal urine were all negative. CONCLUSION: It is feasible to use CAg from schistosomasis patients' urine to prepare anti-CAg McAb by intrasplenic immunization in mice. PMID- 12563836 TI - [Observation on the dynamics of specific anti-GST antibodies in rabbits immunized with recombinant 26 kDa GST of Schistosoma japonicum]. AB - AIM: To investigate the dynamics of the anti-GST antibodies in rabbits immunized with recombinant 26 kDa GST of Schistosoma japonicum. METHODS: The specific antibodies were detected weekly by GST-ELISA in rabbits vaccinated with purified recombinant 26 kDa GST antigen of S. japonicum plus FCA/IFCA; rabbits immunized with 0.85% saline plus FCA/IFCA served as control group. RESULTS: The specific anti-GST antibodies began to increase at week 4 post-immunization, the mean OD value in rabbits vaccinated with recombinant GST could reach 1.03 +/- 0.46 (adjuvant control group only 0.42 +/- 0.04). The experiment was completed at week 65 post-immunization when the mean OD values for specific anti-GST antibody in immunized rabbits were 0.94 +/- 0.26(0.29 +/- 0.16 in the control group). CONCLUSION: Strong anti-GST antibody responses could be induced in rabbits vaccinated with recombinant 26 kDa GST of S. japonicum. PMID- 12563837 TI - [Longitudinal observation on a schistosome-infected snail spot as a source of infection in a mountainous village of Dali city]. AB - AIM: To observe longitudinally the changes in survival and infection rate of Oncomelania snails in a positive snail spot in Yunnan. METHODS: A positive snail spot in a ditch in Shalimuzhuang Village, Dali City, was continually observed on the changes in the density of snail population and natural infection rate of the snails by random environmental sampling for 8 times in 4 successive years. RESULTS: During the 8 times of snail survey, living snails were found relatively stable in the ditch, in which positive snails were obtained in 6 occasions, the average infection rate being 3.99%. CONCLUSION: The density and the positive rate of snails in the mountain region remains relatively stable as a source of human infection. Elimination of the positive snail spots is necessary for controlling schistosomiasis. PMID- 12563838 TI - Pilot study of schistosomiasis control in Poyang lake region. AB - AIM: To perform a longitudinal survey of the control of schistosomiasis in Fanhu Village hyperendemic for schistosomiasis, Poyang Lake region of Jiangxi Province from 1992 to 1996. METHODS: Annual mass chemotherapy and health education were implemented in all investigated individuals, in 1996, artesunate was used in high risk population in transmission season as a prophylactic measure. RESULTS: The prevalence of S. japonicum infection reduced from 26.0% in 1992 to 10.7% in 1994, the intensity of infection (geometric mean of EPG of feces) in residents decreased from 1.92 in 1992 to 0.55 in 1994, and the morbidity indicators such as hepatomegaly and splenomegaly in the cases also have been improved significantly after chemotherapy. In 1995, both the infection rate and intensity of infection of schistosomiasis were recurred from 10.7% and 0.55 in 1994 to 18.4% and 1.04 respectively due to the floods. In 1996, the infection rate reduced to 5.0% and the intensity of infection also decreased to 0.25 due to the additional measure. CONCLUSION: A mass chemotherapy in conjunction with health education and oral artesunate for preventing reinfection in transmission season is effective for the control of schistosomiasis in lake regions. PMID- 12563839 TI - [Construction of DNA vaccine including a chimeric gene encoding Cysticercus cellulosae antigen and porcine interleukin-4]. AB - AIM: To construct a fusion expression vector for DNA vaccine including porcine interleukin-4(IL-4) and antigen cC1 to enhance the protective immunity of Cysticercus cellulosae antigen cC1. METHODS: The cDNA fragments encoding porcine IL-4 and cC1 were amplified respectively by PCR and the fused. The obtained chimeric gene IL-4cC1 contained a synthetic linker of ten amino acids and the sequence surrounding its 5' AUG initiatory codon was changed to optimized translational initiation. RESULTS: Identified by restriction enzyme analysis, an insert fragment of 1.5 kb was demonstrated. It had the same sequence as reported and designed by DNA sequencing analysis. CONCLUSION: A fusion expression plasmid containing porcine IL-4 and cC1 was constructed. PMID- 12563840 TI - [Observation on efficacy of artemether compound against vivax malaria]. AB - AIM: To observe the efficacy of artemether compound against vivax malaria. METHODS: Each artemether compound tablet contains 120 mg benflumetolum and 20 mg artemether. 132 patients with vivax malaria were divided into 3 groups. Group A, 36 patients received 8 tablets as an initial dose, followed by 4 tablets daily for 2 days; group B, 41 patients received 8 tablets as an initial dose, followed by 3 tablets daily for 4 days; group C, 55 patients receiving chloroguine primaquine served as control. Two patients of group A were voluntarily to be bitten by Anopheles dirus before and after medication to observe the influence of artemether compound on the sporogony. RESULTS: The average defervescence times for groups A, B and C were 22.3 h, 23.2 h and 25.0 h (P > 0.05), respectively, the average parasite clearance times were 33.5 h, 30.5 h and 44.9 h, respectively, the average parasite clearance times of groups A and B were all significantly shorter than that of group C (P < 0.01). The replase rates of groups A, B and C were 84.9%, 78.8% and 22.9%, respectively, followed-up at nine months, the relapse rates of groups A and B were higher than that of group C(P < 0.01). CONCLUSION: Two regimens of artemether compound have the advantage of high efficacy against vivax malaria. PMID- 12563841 TI - Advances in experimental studies on albendazole and other drugs against hydatid disease. PMID- 12563842 TI - [Advances in the research of genetic variation of schistosomes]. PMID- 12563843 TI - [Misidentification of Pentastomida parasite in a case of human pentas to miasis in China]. PMID- 12563844 TI - [Survey on filariasis in Yunnan Province]. PMID- 12563845 TI - [Cloning of cDNA encoding asparagine peptidase of Chinese strain of Schistosoma japonicum and its expression in BALB/c mice]. PMID- 12563846 TI - [Distribution of chloroquine in mitochondria, microsome and acid-precipitated protein from Plasmodium berghei]. PMID- 12563847 TI - [Observation on the control of Ascaris, Trichuris and hookworm infection by deworming treatment]. PMID- 12563848 TI - [Achievements in the research on the prevention and treatment of malaria in China]. PMID- 12563849 TI - [Advances in the research on the endemicity and control of schistosomiasis in China]. PMID- 12563850 TI - [Achievements in the mosquito study in China in 50 years]. PMID- 12563851 TI - [Eight-year surveillance of imported malaria in YongKang city]. PMID- 12563852 TI - [Achievements in the research on filariasis in China in the past 50 years]. PMID- 12563853 TI - [Control of an outbreak of vivax malaria in Qiongzhong county]. PMID- 12563854 TI - [Achievements and experiences in the prevention and treatment of clornorchiasis]. PMID- 12563855 TI - [Analysis of 5 cases with cysticercosis of the occipital cisterna major]. PMID- 12563856 TI - [Safety analysis of dust mite allergen for diagnosis and immunotherapy of asthma and rhinitis]. AB - AIM: To make a retrieval investigation of safety in using Dermatophagoides farinae extract in diagnosis and immunotherapy with patients of asthma and rhinitis. METHODS: A questionnaire evaluation of the safety use of D. farinae extract (SMU-Df) during diagnosis and immunotherapy of patients from 1974 to 1995 was carried out. RESULTS: A total of 8,46,342 injections were statistically analysed, among whom 142 systemic adverse reactions occurred involving urticaria 0.82@10000, severe attack of asthma 0.77@10000, anaphylactic shock 0.07@10000 (CL = 1.4-12.0/million), and angioedma 0.02@10000. The time of onset of systemic reactions of immediate type was < 30 min in 32 cases-times, and 1 h and 2 h in 6 cases-times; the time to onset of late response type was 3 h-48 h in 23 cases times with highest incidence of severe asthmatic attacks. The systemic reactions occurred in 18 subjects during skin test, in 96 cases-times during increasing dose phase, and in 14 cases-times during. Among them 6 cases were of anaphylactic shock, and none of it after emergency treatment. The major cause of manifestation of systemic reactions for 41 cases after immunotherapy with SMU-Df extract was due to over dosage errors. CONCLUSION: The application of SMU-Df extract in skin test and immunotherapy of asthma and rhinitis sensitive to mite for the past 22 years in this country indicated that the efficacy and safety have been high. PMID- 12563857 TI - [Cloning and sequencing of the genes coding for the histidine-rich protein II of Plasmodium falciparum]. AB - AIM: To compare and analyze the homology of genes encoding histidine-rich proteinII (HRPII) of different Plasmodium falciparum isolates. METHODS: Using PCR technique, the complete genes coding for HRPII of P. falciparum isolates FCC1/HN and VN isolates were amplified. PCR products were digested by HindIII/BamHI and cloned into plasmid pUC19. The recombinant plasmid HRPII/pUC19 was screened and identified by PCR and restriction analysis. The cloned HRPII genes were sequenced by Sanger's method. RESULTS: HRPII genes of FCC1/HN and VN isolates were successfully amplified and cloned into pUC19. DNA sequencing showed that the coding length of HRPII gene was 1,020 bp without introns in FCC1/HN and VN isolates, however, there were ten points mutations between them. FCC1/HN isolate exhibited 98.8%, 92.2% and 98.7% homology in amino acids with isolates VN, IMTM22, and Itg2, respectively. Though the numbers of repeat sequences were different in four isolates, they had the same hydrophobic leader sequence and a single putative glycosylation site. The secondary structure analysis showed that the main antigenic determinants of four isolates were located on 5' end non repeat region (amino acids 1-60). CONCLUSION: FCC1/HN isolate was highly homologous in the coding region of HRPII with VN, IMTM22, and Itg2 isolate. Four isolates exhibited similar structural characteristics and antigenic determinants in HRPII. PMID- 12563858 TI - [Effect of nitroquine on the membrane phospholipid of intraerythrocytic Plasmodium yoelii in vitro]. AB - AIM: To study the mechanism of antimalarial action of nitroquine. METHODS: Intraerythrocytic P. yoelii was cultured by the method of Trager and Jensen. The amount of [3H]-ethanolamine incorporation was measured as an index of the phospholipid synthesis. DPH was used as a probe to measure the plasmodial fluorescent polarization. RESULTS: The incorporation of [3H]-ethanolamine into the P. yoelii infected erthrocytes was markedly inhibited by nitroquine. The plasmodial membrane polarization and viscosity were significantly increased by nitroquine. CONCLUSION: Nitroquine could inhibit the phospholipid synthesis and decrease the membrane fluidity of P. yoelii. PMID- 12563859 TI - [Sequencing and analysis of a cDNA clone CO111 of Plasmodium falciparum]. AB - AIM: To sequence and analyse a cDNA clone CO111, reacting with immune sera obtained from rabbits immunized with Plasmodium falciparum and one McAb against P. falciparum. METHODS: cDNA clone was amplified by PCR. The PCR product was purified and polished with Klenow enzyme and ligated into the M13mp18 vector (digested by SamI) and then transformed into E. coli JM109. The positive recombinant was screened out by PCR and sequenced by the PRISM Dye Primer Sequencing Kit(ABI). The sequence was analyzed by the program from Geneva University and was compared by GenBank of EMBL. RESULTS: The nucleotide sequence of this cDNA clone contains an open-reading frame of 233 bp, which encodes a predicted polypeptide of 77 amino acid residues. The ratio between A + T and G + C is 3.16. The polypeptide is highly hydrophilic and flexible. Comparison among cDNA of P. falciparum from GenBank of EMBL showed that no sequence identical to this cDNA was found. CONCLUSION: A novel cDNA clone reacted with the antibodies against P. falciparum was isolated. PMID- 12563860 TI - [Studies on immunoprotection in mice after immunization with Schistosoma japonicum 22.6 kDa recombinant protein]. AB - AIM: To evaluate the immunoprotective effect of Schistosoma japonicum recombinant 22.6 kDa (rSj22.6) and Sj22.6/Sj26 GST fusion protein. METHODS: The Sj22.6/Sj26 GST fusion protein was prepared by affinity chromatography using glutathione Sepharose 4B. The purified rSj22.6 could be cleaved easily from the fusion protein with Thrombin. 17 and 12 mice immunized with rSj22.6 and Sj22.6/Sj26 GST separately were each challenged with 40 +/- 1 S. japonicum cercariae. RESULTS: In BALB/c mice, the rSj22.6 and Sj22.6/Sj26 GST could induce 32.1 (P < 0.005) and 34.9% (P < 0.02) worm reduction, respectively, as well as 28.4% (P < 0.02) and 45.1% (P < 0.005) total egg reduction, respectively. CONCLUSION: Bpth rSj22.6 and Sj22.6/Sj26 GST fusion protein are partially effective against S. japonicum. PMID- 12563861 TI - [Effect of liposomal albendazole on the ultrastructure of Echinococcus granulosus cysts in mice]. AB - AIM: To observe the histopathological changes of Echinococcus granulosus cysts in mice treated with liposomal albendazole and co-administration with cimetidine by light microscopy and electron microscopy. METHODS: An oral dose of liposomal ABZ with different formulations was given at 200 mg/kg.d. Cimetidine was administered daily at an oral dose of 100 mg/kg.d. Sixty-seven mice were orally given different drugs six days per week for a total of twelve weeks. RESULTS: The histopathological changes indicated that there were significant differences (P < 0.01) between treated groups and control group. The degeneration and necrosis of E. granulosus cysts were marked in liposomal albendazole combined with cimetidin group. CONCLUSION: Liposomal albendazole was more effective against E. granulosus cyst than albendazole. Cimetidine had an apparent synergistic effect when given in combination with liposomal albendazle. PMID- 12563863 TI - [Dynamic changes in the expression of type VI collagen in mice with schistosomal liver fibrosis]. AB - AIM: To clarify the dynamic changes in type VI procollagen mRNA expression and collagen content in mice with schistosomal liver fibrosis. METHODS: Northern blot hybridization and immunohistochemical technique. RESULTS: The expression of alpha 1(VI) procollagen mRNA measured by Northern blot hybridization in the liver of schistosome infected mice was elevated significantly at 8 wk post infection, peaked at 10 wk pi, slightly decreased at 12-16 wk pi, and sustained at high levels until 20 wk pi. Immunohistochemical assay revealed that the staining pattern of type VI collagen first appeared in the walls of central veins and liver sinusoids at 8 wk pi, then extended gradually to the egg granuloma and the portal tract, within and surrounding the egg granuloma to form dense reticular septum at the 12, 16 and 20 wk pi, respectively. The VI collagen content as indicated by the intensity of stain peaked at 16 wk pi. CONCLUSION: Both the expression of alpha 1(VI) procollagen and the content of collagen VI were significantly increased in the liver of mice with early schistosomal fibrosis, suggesting that the type VI collagen detection might be of importance for evaluating the intensity of schistosomal liver fibrosis. PMID- 12563862 TI - Sequence analysis of the MSP 1 gene of Plasmodium falciparum isolates from Hainan, China. AB - AIM: To obtain the complete sequence and analyze the diversity of the MSP 1 molecule from the Chinese isolates of Plasmodium falciparum. METHODS: Genomic DNA was prepared directly from blood samples spotted on filter papers from 2 malaria patients from Baoting County, Hainan Province. PCR amplification of the target gene was carried out using 5 pairs of oligonucleotides specific for the MSP 1 gene. Direct sequencing of the target gene fragments was performed using ABI PRISM Dye Terminator Cycle Sequencing Ready Reaction Kit (Perkin Elmer) in a automatic ABI PRISM 310 Genetic Analyzer. RESULTS: For the first time, two complete sequences of the MSP1 gene from two Chinese isolates of Plasmodium falciparum were obtained. A comparison with the previously reported sequences identified them as members of the MAD20 allelic family. The deduced amino acid sequences of the MSP1 from this two Chinese isolates were identical with each other except for Blocks 2, 4 and 8. CONCLUSION: The sequences of the MSP 1 from two Chinese isolates of P. falciparum belong to the MAD20 allelic family. Minor variations through the whole sequences exist compared with the MAD20 sequence. The results provide the first evidence of the diversity of the MSP 1 molecule from Chinese isolates of Plasmodium falciparum. PMID- 12563864 TI - [Survey of taenia saginata infection in students in Harbin city]. PMID- 12563865 TI - [Humoral immune response in mice to hybrid nucleic acid vaccines containing Plasmodium falciparum merozoite surface protein 1 block 17-based gene]. AB - AIM: To analyse the humoral immune response in mice to nucleic acid vaccines (VR1012/HG-MSP1-17 for intracellular expression or VR1012/TPA/HG-MSP-17 for secretion) containing Plasmodium falciparum merozoite surface protein 1 (MSP1) 17 block gene and gene fragment of several T cell epitopes from MSA1, MSA2, RESA, IL 1 and TT. METHODS: BALB/c or C57BL/6 mice received three times intramuscular immunization with 200 micrograms/100 microliters or 100 micrograms/100 microliters of VR1012/HG-MSP1-17 or VR1012/TPA/HG-MSP1-17 per mouse each time. Anti-HG or anti-MSP1-17 antibodies were monitored by indirect ELISA. RESULTS: BALB/c and C57BL/6 mice immunized with 100 micrograms/100 microliters of VR1012/HG-MSP1-17 per mouse raised significantly anti-HG and anti-MSP1-17 antibodies, but the levels of antibodies were not high. BALB/c mice immunized with 200 micrograms/100 microliters of VR1012/HG-MSP1-17 per mouse raised higher anti-HG antibodies but not anti-MSP1-17 antibodies. BALB/c mice immunized with 200 micrograms/100 microliters of VR1012/TPA/HG-MSP1-17 per mouse raised low level of anti-HG antibodies only. CONCLUSION: VR1012/HG-MSP1-17 is more immunogenic than VR1012/TPA/HG-MSP1-17. PMID- 12563866 TI - [Studies on allelic zymogram of periodic Brugia malayi from Zhejiang Province]. AB - AIM: To study the allelic zymogram of periodic B. malayi from Zhejiang Province. METHODS: 180 adult worms(male 60, female 120), 32,000 microfilaria and 1,500 infective larvae of periodic B. malayi were examined for the isoenzymes of 14 enzymes by horizontal starch gel electrophoresis with 14 different isomyme. RESULTS: Twenty-seven allelic loci were found in three developing stages of B. malayi, most of them were homozygotes, however, two of them(MPI,MDH) were heterozygotes(7.4%). 17, 19 and 9 loci were presented in adult worms, microfilariae and infective larvae, respectively. At the same time, the allelic zymogram of B. malayi from Guizhou Province was also examined. CONCLUSION: The allelic zymogram of periodic B. malayi from Zhejiang Province was similar to that of periodic B. malayi from Guizhou Province but passaged in the laboratory. PMID- 12563867 TI - [Establishment of a rabbit model of Acanthamoeba keratitis]. AB - AIM: To establish an animal model of Acanthamoeba keratitis. METHODS: Six New Zealand white rabbits were each injected intrastromally with Acanthamoeba suspension 3 days after subconjunctival injection with dexamethasone. RESULTS: All of the 6 rabbits developed keratitis. Acanthamoeba protozoa were identified by the methods of corneal scraping with 10% potassium hydroxide wet mount examined under microscope, corneal protozoa culture and pathological section examination. CONCLUSION: A rabbit model of Acanthamoeba keratitis was established. PMID- 12563868 TI - [Application of expressed sequence tag (EST) in parasite genome research]. PMID- 12563869 TI - [Detection of Cysticercus bovis in a calf injected intravenously with taenia saginata hexacanth]. PMID- 12563870 TI - [Follow-up survey of the hydrocele testis in Changle city where filariasis had been basically eradicated]. PMID- 12563871 TI - [Survey and treatment of clonorchiasis in Fushan city]. PMID- 12563872 TI - [Survey of toxoplasma infection in fertile age women and children in Urumqi city]. PMID- 12563873 TI - [Relationship between hygienic habit and parasitic infection]. PMID- 12563874 TI - [Clinical analysis of 450 cases with liver and abdominal hydatid disease]. PMID- 12563875 TI - [33 children with cerebral paragonimiasis]. PMID- 12563876 TI - [Analyse of the site of lesion in auditory neuropathy]. AB - OBJECTIVE: To analyze the clinic signs, the site of lesion and terminology of a kind of hearing loss which is characterized by speech hearing loss. METHOD: The data of 11 patients were collected and analyzed including the clinic symptoms, pure tone audiometry, acoustic impedance audiometry, auditory brainstem response (ABR), electrocochleogram(ECochG), distortion product otoacoustic emission (DPOAE), CT/MRI. RESULT: All these cases had following contradictory phenomena: Pure tone audiograms were low-frequency hearing loss; ABRs were absent or seriously abnormal, and ABR response threshold was worse than pure tone threshold and click subjective threshold; ECochG could be recorded in some patients; The stapes muscle reflex was absent; DPOAEs were normal or near normal, the average amplitude of low-frequency was higher than normal hearing group, but the average amplitude of high frequency was lower than normal hearing group; The imaging examination did not show any abnormality. CONCLUSION: All data of this group show that the sites of lesion in most patients may be in lower brainstem, but the sites of lesion in some patients may be in the spiral ganglion of cochlea. The research of this disease should be continued. PMID- 12563877 TI - [Comparisons of auditory event-related potential P300 recorded with active and passive modes]. AB - OBJECTIVE: To study the responsibilities and the clinical applications of auditory event-related response P300 recorded with active and passive modes in normal young adults. METHOD: P300s were recorded with active and passive modes, that is, to count and not to count the numbers of target stimuli during measuring respectively, in young adults as subjects. The waveforms, latencies and amplitudes of P300 were analyzed. RESULT: The resemble P300 in waveform and responsibility could be recorded with active and passive mode. There were some differential responsibilities between active and passive recording modes, which the latencies in active mode were shorter than the ones in passive mode, the amplitude of N1-P2 in active mode was lower than the one in passive mode, nevertheless, the amplitude of N2-P3 in active mode was significantly higher than the one in passive mode. CONCLUSION: The resemble P300 in waveform and responsibility could be recorded with active and passive mode. The differences of latencies and amplitudes between two modes demonstrated that the responsibility of P300 was related to the recognition process of auditory center. On the other hand, the passive recording mode of P300 is a way to measure for the uncooperative subjects. PMID- 12563878 TI - [The study of the distortion product emission in the people with hypertriglyceride]. AB - OBJECTIVE: To study the change of the amplitude and latency of the distortion product emission (DPOAE) in the people with hypertriglyceride. METHOD: The DPOAE amplitude and latency of 30 people with hypertriglyceride and 18 normal was acquired with ILO-V5. RESULT: Although the hearing threshold of pure tone was not affected, but DPOAE amplitude of the people with hypertriglyceride was significantly decreased with the normal people, the changes of latency was not significantly affected. CONCLUSION: The cochlea of patient with hyperlipidemia may be damage in its early-stage. The generating mechanism of DPOAE latency and amplitude may be different. PMID- 12563879 TI - [The clinical value of click stimuli on the measurement of ipsilateral acoustic reflex thresholds]. AB - OBJECTIVE: To define that click can substitute for pure tone as a stimulus to measure ipsilateral acoustic reflex thresholds, and to find out the effect of probe frequency on acoustic reflex thresholds. METHOD: Using middle ear analyzer (GSI 33, version 2), we measured 23 normal voluntary participants (46 ears) for getting the pure-tone generated ipsilateral acoustic reflex thresholds at different probe frequency and the click evoked ipsilateral acoustic reflex thresholds at click rate 100/s and 180/s for every ear. RESULT: For pure tone, the ipsilateral acoustic reflex thresholds at 500 Hz, 1000 Hz, 2000 Hz, were (84.35 +/- 6.96) dB, (84.65 +/- 5.93) dB, (87.96 +/- 6.36) dB by using 226 Hz probe; (93.13 +/- 6.54) dB, (92.70 +/- 6.33) dB, (93.35 +/- 5.99) dB by using 678 Hz probe; (86.52 +/- 4.72) dB, (87.48 +/- 5.00) dB, (88.30 +/- 6.16) dB by using 1000 Hz probe. For click, the ipsilateral acoustic reflex thresholds at click rate 100/s and 180/s were (89.78 +/- 6.83) dB, (92.07 +/- 7.42) dB by using 226 Hz probe; (90.44 +/- 6.76) dB, (90.65 +/- 6.38) dB by using 678 Hz probe; (88.04 +/- 6.87) dB, (88.69 +/- 7.42) dB by using 1000 Hz probe. The acoustic reflex thresholds for 678 Hz probe were higher than those obtained with 226 Hz and 1000 Hz probes. CONCLUSION: There were no artifact appearance while stimulus had been click at low rate, so click can be used for the measurement of ipsilateral acoustic reflex thresholds. PMID- 12563880 TI - [Influence of cleftpalate on middle ear conduction and eustachian function]. AB - OBJECTIVE: To explore the influence and its degree of cleftpalate on middle ear conduction and eustachian function. METHOD: 41 cleftpalate patients (82 ears) were performed with otopharyngeal examinations, form of the ostium pharyngeum tubae auditivae observations and acoustic impedance measurements. The results were compared with healthy persons of normal hearing. RESULT: In the cases with cleftpalate, tympanic membrane pathological change incidence was 89.0% (73/82), ostium pharyngeum tubae auditivae of cracked shape was 59.5% (25/42), abnormal tympanogram was 83.1% (64/77), the negative rate of stapedius muscle reflex was 84.4% (65/77). There are significant different in comparison with healthy persons. In synthetic analyses, the rate of secretory otitis media with varied degree in the cleftpalate cases was 74.0% (57/77). CONCLUSION: There was a high incidence of middle ear pathological change and secretory otitis media in the cleftpalate patients. This high incidence was due to susceptivity of the nasopharynx on the infection, weak strength of the dilator tubae eustachii and the deformed shape of the ostium pharyngeum tubae auditivae. PMID- 12563881 TI - [Study on the coagulation state of sudden deafness]. AB - OBJECTIVE: To explore the coagulation, anticoagulation and fibrinolyic activity in sudden deafness (SD). METHOD: The plasma samples from 42 patients with SD before treatment, 23 patients after treatment and 32 normal controls were examined for coagulation factors, anticoagulation, fibrinolyic activity and platelet functions. RESULT: The functions of coagulation and platelet enhanced, the functions of anticoagulation and fibrinolyic system decreased in 42 patients with SD before treatment. We treated with vitamin, dextranum, energetic compound, vasodialator, antiviral ect in 23 patients for a month. The functions of coagulation, anticoagulation and platelet were no significant difference between the after treatment group and control group (P > 0.05). The level of D-dimer obviously elevated (P < 0.01). CONCLUSION: The functions of coagulation and platelet enhanced, the functions of anticoagulation and fibrinolyic system decreased in SD before treatment, The secondary fibrinolyic activity enhanced after treatment. The thrombosis is one of important pathogenesis of SD. PMID- 12563882 TI - [Malformations of chorda tympani nerve]. AB - OBJECTIVE: To analyze the characteristics of anomalies of chorda tympani nerve. METHOD: Chorda tympani nerve and its adjacent structure were investigated during microsurgery in 667 cases of congenitally malformed external-middle ear. RESULT: Anomalies of chorda tympani nerve were found in 433 cases (64.91%). In the group of isolated middle ear malformation of chorda tympani nerve was 16/147 (10.88%); in the group of stenosis of external auditory canal 22/95 (23.15%); and in the group of atresia 394/394 (100%). In 385 cases (97.7%) chorda nerve were not discovered during operations; It were situated at atresia plate (far from malleoincudal interspace) in 9 cases (2.28%). Different feature were found such as: elongation of chorda tympani posterior canaliculus, high or low position, reduplication of chorda tympani nerve, significant low position of chorda tympani nerve in the atresia plate. CONCLUSION: Malformation of chorda tympani nerve is related to the embryologic development of the facial nerve, ossicle, oval window et. It is imparative to identify and to protect the ectopic chorda tympani nerve during operation early. PMID- 12563883 TI - [Analysis on voice acoustic parameters and electroglottographic parameters in normal children of Han, Naxi, Bai nationality]. AB - OBJECTIVE: To explore the normal vocal acoustic parameters and electroglottographic (EGG) parameters and their differences in normal children of the Han, the Naxi, the Bai nationality. METHOD: By using computer and Dr. speech software, We studied six parameters of voice acoustic and the E.G.G. in 951 children (Han 342, Naxi 224, Bai 385), aged 4-8 years old. We also examined 30 cases who suffered from vocal nodule and local proliferation. RESULT: The major parameters 1. Jitter, 2. Shimmer, 3. Mean F0, 4. SD F0, 5. NNE) of the voice acoustic and E.G.G. of the three nationality. CONCLUSION: 1. All parameters in Han nationality school-age children and preschool children were not different in years old. But Naxi and Bai nationality school-age children's Mean F0 and SD F0 were different from preschool children. The parameters(Mean F0, SD F0) of school age children in Han, Naxi and Bai nationality were significant different. 2. The tested parameters were significant different between normal children of the Han nationality and the vocal nodule patients. PMID- 12563884 TI - [Thyroidectomy and injury of recurrent laryngeal nerve]. AB - OBJECTIVE: To study factors related to recurrent laryngeal nerve (RLN) paralysis, a major complication of thyroidectomy. METHOD: Retrospective study of outcome of patient's medical records in Xijing Hospital and Tumor Hospital of Liaoning Province. Records of 1,563 patients who underwent thyroidectomy by departments of general surgery, ENT and head & neck surgery at these two hospitals were reviewed for RLN paralysis. MAIN OUTCOME MEASURES: RLN injury was analyzed in relation to types of surgery, RLN identification, and histopathology. RESULT: The incidence of RLN paralysis was 7.8%, and significantly related to the histopathologic findings of malignancy (P < 0.01). The positive identification and prevention of RLN would not be in either permanent RLN paralysis or temporary RLN paralysis, and the types of surgery were not found to be significant factors in both paralysis. CONCLUSION: RLNs should be identified to avoid iatrogenic injury and subsequent paralysis, and meticulous surgical technique should be applied in patients whose results of biopsy suggested malignancy. PMID- 12563885 TI - [The detection and analysis of human papillomavirus in infantile respiratory tract papillomas by PCR/RFLP study]. AB - OBJECTIVE: To study the relationship between human papillomavirus (HPV) and pathological characteristics of the patients with infantile respiratory tract papillomas (IRP). METHOD: Thirteen cases of infantile respiratory tract papillomas tissue were analysed for Papillomavirus by the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). RESULT: The amplified products of the HPV-L1 region by PCR by using the consensus primers showed that the presence of HPV was 100 percent. Eight cases(61.5 percent) were positive for HPV6 only. Four cases (30.8 percent) were positive for HPV11 only. One case (7.7 percent) showed the presence of HPV18. None were positive for HPV16, 31, 33, 52, 58. CONCLUSION: Infantile respiratory tract papillomas are associated with HPV6, 11, 18 infection, but the subtypes of HPV are not associated with clinical pathological characteristics. PMID- 12563886 TI - [The semiconductor diode laser application in tympanoplasty]. AB - OBJECTIVE: To investigate the method, indications and initial results of semiconductor diode laser application in tympanoplasty. METHOD: The diode laser was used in 14 tympanoplasty procedures. The laser manipulation was mainly used in cases of malleus and incus bony fixation with the tympanum, the fibrous scar tissue around the ossicular chain and the malleus head resection. RESULT: There was no postoperative vertigo and facial paralysis. The average hearing improvement was 24 dB. Seven cases (50%) reached an air-bone gap within 20 dB. CONCLUSION: The application of diode laser in tympanoplasty could effectively reduce the manipulation trauma of ossicular chain and control the bleeding of operative field, especially in managing the bony adhesion of the ossicular chain. PMID- 12563887 TI - [Examination and significance of ABR in non-deafness tinnitus patients]. AB - OBJECTIVE: Research on examination and significance of the auditory brain stem response (ABR) in non-hearing loss tinnitus patients. METHOD: Recording from forehead-mastoid comparing ABR of non-hearing loss tinnitus patients with normal hearing youths. RESULT: In the tinnitus patients, the prolongate latency percentage of wave I was 51.6%; the wave III was 20.9%; wave V was 17.9%. CONCLUSION: Although hearing-loss was not founded in the patients, but the cochlea of some patients had been damaged. PMID- 12563888 TI - [Methylcobalamin, antagonist to transient ototoxic action of gentamicin]. AB - OBJECTIVE: To explore whether is methylcobalamin effective in prophylaxis of AmAn ototoxic action. METHOD: We attempted to use a single injection of gentamicin(125 mg/Kg b.i.w.) to make an animal (guinea pig) model of transient sensory disturbance of hearing, and by a series observation of the changes in ECochG to find out the antagonistic action of methylcobalamin which was simultaneously given to animals(1 mg/Kg, i.m.) followed injection of gentamicin. RESULT: 1. The transient ototoxic action of gentamicin may cause delay of latency and diminution of amplitude of CAP N1 and N2 in animals with a single injection of gentamicin. Though the interval between N1 and N2 prolonged, the changes were still within normal ranges. 2. By contrast, the changes of delay at N1 and N2 as mentioned above were significantly improved in animals administered simultaneously with methylcobalamin. CONCLUSION: It indicated that in addition to the improvement of physiologic function of nerve fibres, methylcobalamin may be also an agent to prevent the transient ototoxic action of gentamicin. Due to methylcobalamin acts as a methyl donor, it participates the synthesis of SAM (S-adenosylmethionine) and thus products GSH(Glutathion SH), the latter may inhibit ototoxic side effects of gentamicin. PMID- 12563889 TI - [The effect of functional endoscopic sinus surgery on mucociliary clearance]. AB - OBJECTIVE: To explore the effect of functional endoscopic sinus surgery(FESS) on the mucociliary clearance of nasal mucosa. METHOD: 105 patients suffered from chronic sinusitis and undergone FESS from Jan. 1998 to Dec. 1998 were staged and analyzed according to the standard set up in Haikou in 1997. The nasal mucociliary transport rate (MTR) was measured by "saccharin" before, three months and six months after surgery. RESULT: 1. The difference of the stages for chronic sinusitis affected directly the surgical result. 2. The difference of properties, limits and extents of pathological changes in sinuses was significantly associated with the MTR. The MTR showed a marked reduction in stage II-II, II-III and III. The MTR was increased after surgery. 3. The difference of the stages affected directly the recovery of MTR. The MTR in stage I-I, I-II, I-III and II-I recovered more quickly. CONCLUSION: The MTR of nasal mucosa tended to recover after FESS. The study confirmed that the present staging system for chronic sinusitis was seasonable. We therefore propose the "saccharin" technique as a simple, safe, reliable and objective method of assessing the results of surgery. PMID- 12563890 TI - [Study of the relationship between sIgE of dust mite in serum and nasal secretion of allergic rhinitis]. AB - OBJECTIVE: To search for the relationship between specific IgE of Dermatophagoides farinae in the serum and that in the local secretion of perennial allergic rhinitis patients. METHOD: The serum and nasal secretion of suspected patients were collected, then test the concentration of mite sIgE in the serum as well as that in the secretion. The results were analyzed at last. RESULT: The relationship of mite sIgE concentration and logarithm of the concentration between the serum and the local secretion is obvious (relative coefficient is 0.675 and 0.846 respectively.) CONCLUSION: Dertermining the Dermatophagoides farinae sIgE in the serum can reflect the local condition of suspected patients fairly well. While for some patients whose local condition is not according to the serum sIgE, testing the local sIgE is necessary to get a correct diagnose. PMID- 12563891 TI - [Effect of intranasal treatment with capsaicin on polyp recurrence after polypectomy and ethmoidectomy]. AB - OBJECTIVE: To evaluate the influence of polypectomy and anterior ethmoidectomy associated with intranasal applications of capsaicin on the recurrence of nasal polyps. METHOD: Fifty one patients (19 females, 32 males, mean age 43) undergoing polypectomy and anterior ethmoidectomy were included in the study. In 29 patients, a cotton pellet soaked with capsaicin (3 x 10(-6) mol, dissolved in 70% ethanol) was left into the middle meatus for 20 min under local anaesthesia. As a control group, 22 patients were treated with the capsaicin vehicle alone (70% ethanol). All patients studied received the intranasal capsaicin or vehicle treatment once a week for 5 weeks. The clinical objective examination and the subjective evaluation of nasal obstruction, rhinorrhea were recorded by means of a visual analogic scale graded from 0 to 4 throughout the study. RESULT: Patients treated with intranasal capsaicin application reported 9 months later and 55 months later separately, a marked reduction of their subjective evaluation of nasal obstruction when compared to the pretreatment evaluation (P < 0.01). In contrast, patients treated with the capsaicin vehicle alone did not have any significant improvement of their subjective evaluation of nasal obstruction. Subjective rhinorrhea was not modified by either treatment. Of 29 patients with capsaicin treatment, 25 (86.2%) patients (9 months later) and 20 (69%) patients (55 months later) showed satisfactory result, compared to 10 (45.4%) patients and 8 (36.4%) patients separately in the control group, the difference was significant (P < 0.01). CONCLUSION: Intranasal capsaicin application can reduce polyp recurrence and nasal obstruction after polypectomy and ethmoidectomy. PMID- 12563892 TI - [Detection of Epstein-Barr virus and human papillomavirus in sinonasal malignant neoplasms]. AB - OBJECTIVE: To investigate the existance of Epstein-Barr virus (EBV) and papilloma virus(HPV) in sinonasal malignant neoplasms. METHOD: EBV and HPV (including HPV6,11,16,18,33) gene were detected in paraffin-embedded tissues with polymerase chain reaction(PCR) from 32 cases of sinonasal malignant neoplasms. RESULT: EBV was detected in 12(37.5%), HPV in 21(65.6%); The coinfection of EBV and HPV was found in 6 cases with stage III-IV by TNM; 10 cases of nasal polyps contained neither EBV nor HPV. CONCLUSION: Like HPV, there may be relationship between EBV and sinonasal malignant neoplasms. The coinfection of EBV and HPV is related to stage III-IV by TNM of sinonasal malignant neoplasms. PMID- 12563893 TI - [Pediatric intranasal and skull base needle endoscopic surgery and examination]. AB - OBJECTIVE: To explore the values of the application of needle endoscope in pediatric nasal cavity and skull base surgery. METHOD: Using a kind of needle endoscope with the diameter of 1.9 mm and digital video image system, 8 cases of pediatric nasal cavity and skull base examination and surgery were performed. RESULT: Of 3 cases of congenital choanal atresia, one improved, the others recovered. One case (2.5 years old) of traumatic cerebrospinal fluid (CSF) fistulae was successfuly repaired under the 1.9 mm needle endoscope. There were 2 cases of congenital meningoencephalocele. One was 1 month newborn. When the diagnosis was confirmed by needle endoscopy and CT scan, the operation was postponed. The other got endoscopic operation and had recovered. Of 2 cases of skull base malignant tumour, the diagnosis was confirmed by intranasal biopsy under needle endoscope. CONCLUSION: With the application of needle endoscope, the pediatric skull base surgery and examination can be performed in the narrow nasal cavities and sinuses with less invasion and good result. PMID- 12563894 TI - [The study of the nasal airway reactivity in specific nasal provocation test]. AB - OBJECTIVE: To explore efficient evaluation method for specific nasal provocation test(SNPT) and to observe the pathogenic process and patho-physiological changes of allergic rhinitis. METHOD: 30 patients with perennial allergic rhinitis(patients group) and 14 normal adults(controls group) were challenged unilaterally with the Coca's solution and/or the solution for allergen extracts. The nasal resistance variations in all subjects were monitored with active anterior rhinomanometry and the nasal symptoms were observed. RESULT: The symptoms of nasal stiffness, sneezing and rhinorrhea appeared in most patients between 0.5 and 3 minutes after provocation. The ratio of continual sneezes above 5 in patients group(83.3%) was significantly higher than in controls group (21.4%) (P < 0.01). The increased ratio of total nasal resistance and unilaterally-challenged nasal resistance were significantly higher in patients group than in controls group (P < 0.05) and no difference between coca's solution in patients group and controls group. CONCLUSION: Positive SNPT was indicated if obvious nasal symptoms appeared and the unilaterally-challenged nasal resistance increased 100% as well as the total nasal resistance increased 40% after SNPT. PMID- 12563895 TI - [Treatment of diseased middle turbinate in endoscopic sinus surgery]. AB - OBJECTIVE: To study the clinical significance of reservation of the middle turbinate in functional endoscopic sinus surgery (FESS) for sinusitis and/or nasal polyps. METHOD: According to anatomic criteria of the middle turbinate surgery in adult, 32 cases who had sinusitis and/or nasal polyps were treated and followed up. The recovery of middle turbinate contour, postoperative ethmoid cavity and the rate of patency of the maxillary sinus ostium were observed postoperatively. RESULT: 1. After 6 months, the form of middle turbinate returned to normal in 24 cases (75%), adhesions occurred in 8 cases (25%), including the closure of ethmoid sinus cavity in 2 cases. 2. The patency of maxillary sinuses ostium maintained well in 25 cases (78.1%), stenosed in 6 cases (18.8%) and closed in 1 case (3.1%). The diseased middle turbinate can recover to normal contour in most cases, often plastics in FESS. CONCLUSION: Reservation of middle turbinate plays an important role in promoting the clinical cure rate. PMID- 12563896 TI - [Timing changes of apoptosis and proliferating cells nuclear antigen after intra arteiral infusion chemotherapy for nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the timing changes of apoptosis (APO) and PCNA after intra-arterial infusion chemotherapy for nasopharyngeal carcinoma. METHOD: Ten patients were subjected to percutaneous superficial temporal artery catheterization and infusion of anticancer drugs: 5-Fu, DDp, BLM, nicotinamide. The biopsy of nasopharyngeal tumor tissues was performed before chemotherapy and 7 days after chemotherapy. Apoptotic cells were examined by TOT-mediated DUTP fluorescein and labeling. The expression of proliferating cells nuclear antigen (PCNA) was detected with immunohistochemical staining. RESULT: AI of nasopharyngeal cancer cells before chemotherapy and 7 days after chemotherapy was 0.52% and 1.42%, PI was 42.4% and 51.2%. CONCLUSION: Intra-arterial infusion chemotherapy not only induced apoptosis effectively, but also inhibited temporarily tumor cells proliferation in patients. The curative surgical treatment should be performed as soon as possible after chemotherapy. PMID- 12563897 TI - [Usefulness of CT scan in secretory otitis media in patient with nasopharyngeal carcinoma]. AB - OBJECTIVE: To explore the usefulness of CT scan in nasopharyngeal carcinoma (NPC) with secretory otitis media (SOM). METHOD: CT of the nasopharynx and mastoid in 127 cases with NPC pro- and post-irradiation were analyzed. RESULT: Total incidence rate of SOM was 59% before irradiation. The incidence corresponded to T stage and tumor location of NPC. In the patients without SOM before irradiation, 70 percent of the ears developed SOM, and the peak stage of the formation of SOM was in three to six month after irradiation. CONCLUSION: CT scan for the patient with NPC is useful for the diagnosis and prognosis of SOM induced by NPC. PMID- 12563898 TI - [Voice rehabilitation of modified Amatsu operation after total laryngectomy]. AB - OBJECTIVE: To evaluate voice rehabilitation of one stage tracheoesophagus shunt operation after total laryngectomy. METHOD: One stage voice rehabilitation of 20 cases for post total laryngectomy was performed with modified Amatsu tracheoesophgeal shunt operation. RESULT: 16 cases developed speech with voice rehabilitation among 20 cases of postlaryngectomy. CONCLUSION: One stage tracheoesophageal shunt voice rehabilitation applie to all patients with total laryngectomy and with unilateral or bilateral neck dissection at the same time of total laryngectomy and with radiation therapy. The technique is easy and safe. It can resolve the problem of restoration of voice. PMID- 12563899 TI - [The study of facial neurapraxia]. AB - OBJECTIVE: To study the clinical characteristics and mechanism of facial neurapraxia. METHOD: Patients were tested with nerve excitability method and judged the degree of facial paralysis. Animal mode of neurapraxia was made and threshold of evoked electromyographic potential was tested. The fibers of facial nerve were observed under transmission electromicroscope. RESULT: Average restoration time of evoked electromyographic potential was 52.5 min. The lamellar separation of myelin sheath of facial nerve was found under electromicroscope. Patients with facial neurapraxia was judged as denervation negative and residual function of facial muscles was above 65%. CONCLUSION: Facial neurapraxia is mild reversible facial paralysis and relevant to the lamellar separation. PMID- 12563900 TI - [Observation of deaf-mute students' audition examine and otacoustic effect]. AB - OBJECTIVE: To explore the state of the remnant hearing of deaf-mute students and to check the effect of hearing aids to them. METHOD: 38 cases of deaf and dumb students were examined with the pure tone auditometry, acoustic immitance test, auditory brainstem response, and the environmental sound discrimination test. RESULT: In 17 cases, the average threshold of air conduction of speech frequency is below 89 dB HL; In 18 cases, it is between 90 dB and 110 dB; In 3 cases, it is above 110 dB. 33 cases have model A curve and 5 cases have model B curve. When 123 dB peSPL was presented, 24 cases generated the waveform but the others didn't. 16 cases could discriminate the environmental sound and the other 22 cases could not, but if the patients weared the hearing aids, the number of the persons who could discriminate the environmental sound increased to 24, only 14 cases had no effect by this means. CONCLUSION: Examination of deaf-mutes audition can find persons with residual audition. If they wear the hearing aids and go into hearing rehabilitation training at the same time as early as possible, it will improve the environmental sound discrimination rate. PMID- 12563901 TI - [Evaluation of the effects of the ostiomeatal complex corrective surgery]. AB - OBJECTIVE: To study the effects of the ostiomeatal complex corrective surgery (OCRS) for finding out more suitable surgical style for the nasal obstructive diseases. METHOD: 135 cases treated with OCRS were summed up and analyzed. RESULT: The effect is marked by comparison with the traditional surgery such as middle turbinectomy, ethmoidectomy etc. CONCLUSION: The developing direction of the nasal surgical treatment ought to recover the nasal physiological structure. The OCRS makes great efforts in this direction and the achievements have been confirmed. PMID- 12563902 TI - [Culture of human nasal respiratory epithelial cells in serum free medium supplemented with hormones and growth factors and measurement of human nasal ciliary motility using videomicroscopy]. AB - OBJECTIVE: To establish a culture model of human nasal respiratory epithelial cells and a method of measuring human nasal ciliary motility. METHOD: The human nasal respiratory epithelial cells were detached with collagenase and cultured in serum free medium supplemented with hormones and growth factors, the ciliary beat frequency was measured by videomicroscopy. RESULT: After inoculation, cells cultured with this method adhered in 24 hours, confluented in 6-8 days and lived for 16 days. During that time ciliary beating was active, both acidic and neutral mucoitin granules were rich in goblet cells and all chromosome of 23 pairs were normal, the ciliary beat frequency in 29 subjects' nasal mucosa was (411 +/- 24) beats/min (mean +/- s). CONCLUSION: A culture model of human nasal respiratory epithelial cells in serum free medium supplemented with hormones and growth factors and a method of measuring human nasal ciliary motility was successfully established. PMID- 12563903 TI - [The effect of noise exposure on the expression of NOSmRNA in the cochlea]. AB - OBJECTIVE: To study the effect of NO in noise-induced deafness. METHOD: The effect of noise exposure on the expression of NOSmRNA in the guinea pig cochlea was observed with NADPH-diaphorase histochemistry staining, NOSmRNA hybridization in situ and Northern blotting analysis. RESULT: NADPH-diaphorase histochemistry indicated that NOS mainly presents in the inner hair cells, outer hair cells of Corti's organ, spiral ganglion cells and the marginal cells of stria vascularis in the guinea pig cochlea; NOSmRNA hybridization in situ showed that there is positive signal in the inner hair cells, outer hair cells and spiral ganglion cells, but there is no positive signal in the stria marginal cells of vascularis; Northern blotting showed that the expression of noise exposure group is more intensive than that of the control group. CONCLUSION: There is a Glu: NMDA receptor, NO: cGMP pathway in the cochlear afferent possibly; noise exposure induced overproduction of NO mediates the effect of neurotoxicity; the marginal cells of stria vascular produce and release NO to regulate the microcirculation of the cochlea. PMID- 12563904 TI - [Anatomic study on the nervous distribution of the human recurrent laryngeal nerve]. AB - OBJECTIVE: In order to supply the anatomic data for the selective reinnervation operation of the recurrent laryngeal nerve, the nervous distribution of the recurrent laryngeal nerve was investigated. METHOD: 50 total fresh human larynx were systematically dissected by the method of microdissection. RESULT: In most cases, the recurrent laryngeal nerve diverges at the level of 1.0-2.5 cm under the cricothyroid articulation for forming the extralaryngeal division. There are three types of extralaryngeal division. Except the cricothyroid muscle, all the laryngeal intrinsic muscles are innervated by the anterior branch of the recurrent laryngeal nerve. In 88% of the cases, the posterior cricoarytenoid muscle is innervated by two nervous pedicle, the others are innervated by a single nervous pedicle. CONCLUSION: At the level of the posteroinferior part of the thyroid cartilage, by opening a small window, we can easily find the abducent branch of the recurrent laryngeal nerve, following this branch, we can see the adducent branch and the thyroarytenoid branch. The anatomic specifity of the nervous distribution can supply us the data for the operation of selective nervous reinnervation to overcome the possibility of syncinesis of the laryngeal muscle. PMID- 12563905 TI - [Telomerase activity and expression of its RNA component in laryngeal cancer]. AB - OBJECTIVE: Expression of telomerase activity and telomerase RNA component have been reported in various human cancers. In the article, we ascertained whether telomerase activity and hTR changes are characteristic of laryngeal cancer. METHOD: Telomerase activity were examined by telomeric repeat amplification protocol (Trap) and hTR examined by in situ hybridization, in tissue samples from 50 laryngeal cancer, 12 laryngeal polyps, 14 precancerous, 9 normal laryngeal tissue. RESULT: 1. All tissue expressed various levels of hTR. Positive rate of hTR increased with the clinical stage, but no significant correlation was observed, as well as lymph node metastasis. 2. The telomerase activity were detected in samples 39 of the 50 laryngeal cancer(78%). However, in 12 cases of begin lesions only 8.3% (1/12). Telomerase activity was found to have a relationship with the grade of tumor differentiation. Poorly differentiated laryngeal cancer expressed high level telomerase activity. CONCLUSION: Telomerase activity were an early event of laryngeal cancer. It suggests that detection of telomerase activity could be a good diagnostic marker for distinguish cancer from normal cell. PMID- 12563906 TI - [Quantitative assay of telomerase activity in head and neck squamous cell carcinoma]. AB - OBJECTIVE: To detect and quantify telomerase activity in tissues from head and neck squamous cell carcinoma (HNSCC), and to explore whether the levels of telomerase activity can be a useful marker for cancer risk assessment in HNSCC. METHOD: The levels of telomerase activity of 55 fresh specimens obtained from 25 patients with HNSCC were detected by the method of telomeric repeat scintillation radioactive count, including 25 primary tumors, 7 samples of neck metastases and 23 corresponding normal tissues as control. In 7 patients, both primary and metastatic tumors were obtained. The levels of telomerase activity were determined with a liquid scintillation counter. The basic principle of the technique is that a G-rich oligonucleotide strand of telomeric sequence is used as primer, 3H-dTTPs are incorporated into the products while telomerase elongates the primers, and then free 3H-dTTPs are removed by rinsing step, finally radioactive count per minute (cpm) of products is detected, and the levels telomerase activity can be evaluated according to the value (cpm). RESULT: 1. The levels of telomerase activity (cpm) in tissues with HNSCC were significantly higher than those in the normal tissues (P < 0.05); 2. The levels of telomerase activity in the primary tumors were higher than those of their neck metastases, but the difference was not statistical significance (P > 0.05); 3. The levels of telomerase activity in the neck metastases were remarkably higher than those in the normal tissues (P < 0.05). CONCLUSION: 1. Detection of telomerase activity in HNSCCs can be a useful marker for cancer assessment. 2. To quantify telomerase activity may have clinical diagnostic value for HNSCC. 3. The levels of telomerase activity in the metastatic lymph nodes were higher. PMID- 12563907 TI - [Effects on vestibular function by irradiation in the patients with nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate effects on vestibular function by irradiation in the patients with nasopharyngeal carcinoma. METHOD: Twenty patients with nasopharyngeal carcinoma were routinely irradiated with dose of 70 Gy (3 cases > 70 Gy). RESULT: Nine cases (45%) were abnormal in vestibular function test after radical irradiation in 20 cases (spontaneous nystagmus n = 3; unlateral canal paresis n = 4; bilateral canal paresis n = 2; bilateral vestibular function absent n = 2). The other 11 cases(55%) were normal in vestibular function test. CONCLUSION: Vestibular function may be damaged after radical irradiation in patients with nasopharyngeal carcinoma. During professional identification, this factor must be respected and vestibular function, cochlear function and tubal function must be separately evaluated. PMID- 12563908 TI - [The treatment of nasal adhesion after radiotherapy in the patients with nasopharyngeal carcinoma]. AB - OBJECTIVE: In order to find a best way for treating the nasal adhesion after radiotherapy in the patients with nasopharyngeal carcinoma (NPC). METHOD: 4 cases of nasal adhesion after radiotherapy in the patients with NPC were operated by the way through palate. 32 cases were operated under endoscope, in them, 26 cases were set a tube in nasal cavity after operation, and the other 6 cases were not. And the postoperative consequences were analyzed. RESULT: The curative effect of the reconstruction of nasal cavity with a tube through palate are favorable, but the processes are too complex and the trauma are too large for patients. Whereas the way which only sever the nasal adhesion under endoscope is simple, but the disease recur easily. The reconstruction of nasal cavity with a tube under endoscope not only predigest the operation, but also obtain the well effect. CONCLUSION: The reconstruction of nasal cavity with a tube under endoscope is a good way to treat nasal adhesion. PMID- 12563909 TI - [Assay of Epstein-Barr virus in nasopharyngeal tissues and serum of patients with nasopharyngeal carcinoma]. AB - OBJECTIVE: Epstein-Barr virus (EBV) is associated with nasopharyngeal carcinoma (NPC). Determination of EBV-VCA-IgA, EBV-EA-IgA in serum and assay of EBV DNA (PCR) in biopsy tissue has been used for diagnosis of NPC. This paper evaluated the value of the three determinations for diagnosis of NPC. METHOD: 146 patients were investigated. For each patient determination of EBV-VCA-IgA, EBV-EA-IgA in serum and EBV-DNA (PCR) in biopsy tissue were performed with double-blind studies, of 146 patients, 76 were NPC and 70 non nasopharyngeal carcinoma as control, identified with histopathological examination. The differences of the determinations between NPC and control were compared statistically with chi 2 test. RESULT: Of 76 NPC, the positive rates of EBV-DNA (PCR), EBV-VCA-IgA and EBV EA-IgA were 90.8%, 75.0% and 26.3% respectivelly. For 70 control cases, the positive rates 2.9%, 38.6% and 2.9% respectivelly (P < 0.005). CONCLUSION: Our results suggested that assay of EBV-DNA (PCR) has better sensitivity and specificity than EBV-VCA-IgA and EBV-EA-IgA for diagnosis of NPC. PMID- 12563910 TI - [Immunohistochemical study of the sinus mucosa during chronic maxillary sinusitis]. AB - OBJECTIVE: To study the protective role of some immunocompetent cells and some humoral immune factors in the local mucosa during chronic sinusitis. METHOD: Using immunohistochemical streptavidinbiotin peroxide complex (SABC) method to investigate the distribution and the numbers about the T-lymphocytes and subsets, B-cells, macrophages, NK cells, mast cells eosinophils, HLA-DR antigen and IgA, IgG, complement C3, lysozyme in the local mucosa of two type chronic sinusitis. RESULT: The numbers of T-lymphocytes, T helper/inducer cells, B-cells and macrophages in the mucosa of chronic sinusitis type 1 was significantly more than these in the healthy control (P < 0.01). The number of T-lymphocytes, T suppressor cells, B cells, mast cells and eosinophils in the type 2 was significantly more than these in the healthy control (P < 0.01). The ratio of the helper cells to T suppressor cells significant different in the two type chronic sinusitis. CONCLUSION: The study showed that the infiltrating of immunocompetent cells was different between the two type chronic sinusitis. PMID- 12563911 TI - [Revision endoscopic sinus surgery for recurrent nasal polyps]. AB - OBJECTIVE: To study the surgical effect and experience of revision endoscopic sinus surgery for the patients with recurrent nasal polyps. METHOD: From Jul. 1997 to Oct. 1998, a subset of 21 cases with recurrent nasal polyps had been operated by the way of revision endoscopic sinus surgery. All the patients were operated bilaterally at the same time. The operation type included: total ethmoidectomy or sub-total ethmoidectomy, maxillary natural orifice enlargement and maxillary sinus pathological changes tissue eliminated by canine fossa access. RESULT: We have followed these patients for 6-12 months and evaluated surgical effect: 7 cases of 21 were cured, 8 were improved, 6 were of no effect, and the total effective rate was 71.4%. There are not any heavy complications in this subset. CONCLUSION: Revision endoscopic sinus surgery is an effective way for recurrent nasal polyps patients, six anatomic marks proposed by May will do good for us to resect the pathological changes tissue thoroughly and prevent heavy complications as could as possible. PMID- 12563912 TI - [Bacteriological analysis of chronic sinusitis in school-age children (36 cases report)]. AB - OBJECTIVE: To research the bacteriological characters of chronic sinusitis in school-age children and to provide basis for clinical treatment. METHOD: The purrent discharges taken from middle meatus of 36 patients were cultured for bacteria and the antimicrobial suscepility was determined. RESULT: The positive rate of bacteria culture was 83.3%, the compound infection rate was 36.1%. Aerobic bacteria were present in 76.7%, anaerobic bacteria were present in 55.6%. The result of antimicrobial susceptibility test was scattered. CONCLUSION: Infection rate of anaerobic bacteria was high in chronic sinusitis in school-age children and result of antimicrobial susceptibility was scattered. It was important to have anti-anaerobic treatment in chronic sinusitis in school-age children and to take an antimicrobial susceptibility test before medical treatment. PMID- 12563913 TI - [A study on the cranium internal facial nerve root comb scraping for the treatment of facial spas]. AB - OBJECTIVE: Studying about the good qualities of the treatment for facial spas by cranium internal facial never root comb scraping. METHOD: After improve sigmoid sinus carding operation and several stratums of nerve root being carded, use fine knife to card obliquityly the small never finbers one by one again, part of the small fibers can be scraped off if the spasm is serious. RESULT: One recurrence ocurred after three years' follow-up survey of 23 patients who have received the operation. CONCLUSION: The good quality is "double insurance", increasing curative effect, decreasing the rate of recurrence and preventing the complication. PMID- 12563914 TI - [Diagnosis and treatment of sphenoid sinus cyst]. AB - OBJECTIVE: To better understand the sphenoid sinus cyst and make correct diagnosis and treat it efficiently. METHOD: 47 patients with sphenoid sinus cyst were investigated. Presenting symptoms, radiographic appearances and treatment were analyzed, respectively. RESULT: The present study demonstrated that the incidence of sphenoid sinus cyst had apparently increased in recent years. The most common symptom was headache, accident rate was 64%, followed by ocular disturbance, nose was involved in 15% patients. The optic nerve is the most frequently involved cranial nerve it makes decompression surgery necessary to recovery visual loss. Radiographic finding made contribution to location the lesion and exposition nature of the lesion. With the help of nasal endoscope, more information could be get to establish correct diagnosis, and operation under nasal endoscope would be safe with minimal blood loss, reduced operating time, less pains and short postoperative hospitalization. CONCLUSION: CT, MRI and nasal endoscope provide the greatest diagnosis information and guide management, nasal endoscope plays an important role in treatment of sphenoid sinus cyst. Emergency surgery should be performed on patients with visual loss. PMID- 12563915 TI - [Microwave treatment of ethmoidal nerve under nasal endoscopy in the patients with allergic rhinitis]. AB - OBJECTIVE: To study the effect of microwave treatment of ethmoidal nerve on allergic rhinitis. METHOD: 106 cases of allergic rhinitis patients were treated with ethmoidal nerve microwave under nasal endoscope. RESULT: It showed marked effect in 89 cases(84.0%), immprovement in 12 cases(11.3%) and failed in 5 cases (4.7%) in one year follow-up. The overall effective rate was 95.3%. CONCLUSION: This result indicated that the microwave tissue coagulation in treating allergic rhinitis unde nasal endoscope is a good method for clinical treatment. PMID- 12563916 TI - [Effects on fiber regeneration following facial nerve transection treated with NOS antagonist]. AB - OBJECTIVE: To elucidate the role of nitric oxide synthase antagonist, N-Nitro-L Arginine Methyl Ester (L-NAME) in the regeneration of facial nerve following facial nerve transection. METHOD: The facial nerves of 40 guinea pigs were exposed and a segment of the nerve at 3 mm length were excided. The two ends of the transected nerve were sutured and enveloped in a silicone tubule to form a regeneration chamber. L-NAME and saline were respectively injected into the regeneration chambers just after transection. The experiments were observed quantificational by the means of axon and myelin staining under light microscope. RESULT: The findings in the work indicate that inhibition of NOS with L-NAME substantially better myelinated facial nerve regeneration than that associated with administration of saline. CONCLUSION: L-NAME improve the regeneration of myelinated fibers. The mechanism seems to be related to the blockade of NO's neurotoxicity. PMID- 12563917 TI - [The effect of stress-relaxation plate fixation on the remodeling of the cortex under plate]. AB - OBJECTIVE: To explore the influence of stress-relaxation plate(SRP) fixation on the remodeling of cortex under plate. METHODS: Twenty-eight New Zealand rabbits were used in this study, the bilateral tibia were osteotomized in the middle and fixed with SRP (experimental group) and rigid plate (control group) respectively. The scanning electron microscopy was used to observe the bone remodeling process from 2 to 48 weeks after operation. RESULTS: There was cortex osteoporosis beneath plate in different degree in both experimental and control groups before 8 weeks, it showed as the disorganization of collagen fiber structure and formation of resorption cavities. In comparison, the osteoporosis degree in experimental group showed milder than that of the control group. After 12 weeks, the resorption cavities became smaller, and the structure of collagen fibers became regular with the alignment parallel to the long axis of cortex. In contrast to the experimental group, the bone osteoporosis under plate of control group exacerbated continuously. CONCLUSION: Without removal of the bone plate, SRP fixation not only reduce the degree of plated bone osteoporosis, but also make the osteoporosic bone return to normal. PMID- 12563918 TI - [Bone and joint revision surgery after limb salvage procedure of malignant bone tumor]. AB - OBJECTIVE: To analyze the indications for revision surgery after limb salvage procedure of malignant bone tumor and summarize the experiences in revision surgery. METHODS: From January 1994 to December 1997, 8 cases were re-operated after primary limb salvage procedure. The average survival period with no-tumor occurrence was 8 years. The common causes for the revision were traumatic osteoarthritis, fracture, and bone resorption. The main difficulties in revision were soft tissue contracture and limb discrepancy from limb shortening. RESULTS: In this study, there was total hip replacement in 1 case, large segmental allograft for reconstruction of distal femur in 3 cases, total knee replacement upon composite of previously transplanted allograft in 3 cases, removing of intramedullary nail and re-internally fixed with intramedullary nail in 1 cases. The isotopic bone scan before the revision showed active bone metabolism in all 4 transplanted segmental allograft. The pathologic study of the transplanted allograft after revision confirmed new bone formation in allograft. The revision procedure reduced the pain, and improved the limb function. CONCLUSION: The main causes of revision surgery after limb salvage procedure of malignant bone tumor are fracture of transplanted allograft segment or devitalized tumor segment, and poor function of the affected joint. Constrained knee prostheses with rotating hinges or semi-constrained ball-axis resurfacing knee prostheses improve the function of knee joint postoperatively. PMID- 12563919 TI - [The application of transposition of the gastrocnemius muscle in the limb-salvage operations of the proximal tibial tumors]. AB - OBJECTIVE: To evaluate the efficacy of transposition of the gastracnemius muscle in the limb-salvage operation of the proximal tibial tumor. METHODS: From 1995, transposition of the gastrocnemius muscle was applied to cover the devitalized bone, bone cement or autologous bone graft in 15 cases with tumors of the proximal tibia (transposition of medial heads of gastrocnemius muscle in 12 cases, and lateral heads of gastrocnemius muscle in 3 cases respectively). Among them, there were 7 osteosarcomas, 5 giant cell tumors, 1 malignant fibrous histocytoma, 1 chondrosarcoma and 1 osteoblastoma. The operations included segmental devitalization with 95% alcohol in 7 cases, knee reconstruction of the unilateral tibial plateau with iliac graft in 5 cases, segmental devitalization with microwave in 2 cases, local resection and bone graft in 1 cases. RESULTS: Apart from 2 cases whose wounds needed suturing again due to the liquefaction of the subcutaneous fat around the incision, no wound complications were occured in other 13 patients. No significant loss in the function of the leg and ankle was observed after transposition of the gastrocnemius muscle. There was no local recurrence, but 3 patients died due to lung metastases. CONCLUSION: Transposition of the gastrocnemius muscle after resection of promixal tibial tumors can improve the local blood supply, cover the deep structures and prevent from the failure of limb-salvage operation due to wound complications. PMID- 12563920 TI - [Study on adriamycin-porous tricalcium phosphate ceramic drug delivery system and its drug release test in vivo]. AB - OBJECTIVE: To manufacture adriamycin-porous tricalcium phosphate (A-PTCP) ceramic drug delivery system (DDS) as a possible method for bone defect treatment after bone tumor operation. METHODS: A-PTCP DDS was made from putting adriamycin into PTCP. Thirty rabbits were divided randomly into group A(24 rabbits) and group B(6 rabbits). A-PTCP was implanted in the greater trochanter of the right femur in group A. Adriamycin were injected into veins in group B. Muscle around A-PTCP and plasma were taken out at different period. Adriamycin concentrations in muscle and plasma were measured by high performance liquid chromatography (HPLC). RESULTS: A-PTCP could gradually release adriamycin over 10 weeks. Adriamycin concentrations in the muscle were higher than that in plasma. CONCLUSION: A-PTCP may be a new method for repairing bone defects after bone tumor operation. PMID- 12563921 TI - [Repair of bone defect with benign tumor by embryonic bone transplantation]. AB - OBJECTIVE: To evaluate the clinical results of repair of bone defect by embryonic bone transplantation. METHODS: From January 1994 to June 1999, 148 cases of bone defect were repaired by embryonic bone transplantation following alcohol treatment, there were 63 cases with bone cyst, 42 cases with fibrous dysplasia of bone, 26 cases with giant cell tumor of bone, and 17 cases with enchondroma among them. The maximal bone defect was 3.5 cm x 10.0 cm, while the minimal defect was 0.5 cm x 1.0 cm. RESULTS: All of those bone defect with benign tumor were bone union used by embryonic bone transplantation after 3 months to 1 year of operation, the average healing course was 6.2 months, followed up 1 to 6 years, averaged 14 months, no tumor recurrence and no obvious local or system response were observed. CONCLUSION: Embryonic bone can be used as a good repairing material of postoperative bone defect of benign tumors, the clinical results are satisfactory. PMID- 12563922 TI - [Ectopic osteogenesis of bone marrow stromal cells induced by bone morphogenetic protein]. AB - OBJECTIVE: To investigate the ectopic osteogenesis of bone marrow stromal cells (MSC) induced by bone morphogenetic protein(BMP) in vitro and in vivo, providing the experimental evidence for making an artificial bone with its own capacity of bone formation. METHODS: MSC were separated and cultured from bone marrow of Wistar rats, MSC were co-cultured with BMP in vitro (cultured in plate and diffuse chamber). Artificial coral hydroxyapatites (CHA) with MSC and BMP were implanted into dorsal muscles of Wistar rats, their bone formation were observed by morphological examination, histochemistry and immunohistochemistry. RESULTS: Only cartilaginous matrix were produced by MSC in vitro (cultured in plate and diffuse chamber), and both cartilaginous and bone matrix production within the combined grafts were seen. The bone formation of experimental groups (CHA + BMP + MSC) was stronger than that of control A(CHA + MSC) and control B(CHA). CONCLUSION: It may be possible to produce an artificial bone with its own capacity of bone formation by combined graft (CHA + BMP + MSC). There may be multiple factors as well as BMP inducing bone formation both in the whole body and the location of the implantation. Further research on these factors will have the significance for making the ideal artificial bone. PMID- 12563923 TI - [A new method of bone defect repairing after bone cyst curettage]. AB - OBJECTIVE: To introduce a new method of bone defect repairing after bone cyst curettage. METHODS: Eight cases with bone cyst were treated with this new method. The pieces of autogenous periosteum were implanted into the hematoma within the enveloped bone defect created after the bone cyst curettage. Among these patients, there were 5 males and 3 females, aged from 14 to 36 years old. All the lesions located in the upper of femur except one being located in humerus. The results were evaluated through the postoperative radiological findings with the preoperative ones and analysis of clinical functions. RESULTS: All the patients were followed up for 2 to 11 years. X-ray films showed that osteogenesis developed well and that the enveloped bone defects had been repaired. No recurrence was found and the function of the affected limbs were maintained. CONCLUSION: Autogenous periosteum grafting is effective in the treatment of solitary bone cyst. PMID- 12563924 TI - [Autogenous bone marrow graft for the management of nonunion of tibia]. AB - OBJECTIVE: To investigate the effect of percutaneous bone marrow graft for the management of nonunion of tibia. METHODS: From March 1996 to June 2000, 56 cases with nonunion of tibia were treated by autogenous bone marrow graft. Among them, there were 38 males and 18 females, aged from 19 to 72 years. A marrow needle was inserted into the site of the nonunion under the X-ray, the autogenous bone marrow was injected into the site of nonunion. Compression bandage and appropriate immobilization material were applied after operation. This procedure was repeated every month, 2 or 3 times was needed. RESULTS: 56 patients were followed-up for 4 months to 4 years and 2 months, averaged 2.8 years. Fracture healed in 53 cases and X-ray displayed fracture line disappeared and a great deal of continuous callus formed, and nonunion in 3 cases. CONCLUSION: Percutaneous autogenous bone marrow graft can play a role in osteogenesis at the site of nonunion. It is easy to aspirate bone marrow and the operation is simple. It has clinical application value for the satisfactory effect. PMID- 12563926 TI - [Bone union of distracted region after limb lengthening]. AB - OBJECTIVE: To investigate the factors which affect the bone union of distracted region after limb lengthening, so as improve the curative effect and diminish the incidence of complication. METHODS: To look up the latest literatures dealing with the bone union in limb lengthening, then review the procedure of osteogenesis and the affecting factors. RESULTS: The osteogenesis of distracted region after limb lengthening is a sophisticated procedure. It can be affected by the velocity of lengthening, the period of lengthening, the site and method of osteotomy, the age etiology of patient. CONCLUSION: The bone union of distracted region after limb lengthening can be facilitated by following factors: 1. the velocity of lengthening slower than 1.0 mm/day; 2. moderate delay in distraction; 3. axial shortening of distracted region; 4. micromovement stimulation. PMID- 12563925 TI - [Anatomical study and clinical application of medial femoral condylar bone flap pedicled with the saphenous vessels]. AB - OBJECTIVE: To investigate a new operative method for repairing nonunion and defect of tibia. METHODS: First, observe the relation between the joint branches of descending genicular artery and the saphenous branch of descending genicular artery in 10 cadevers. Then the medial femoral condylar bone flap pedicled with the saphenous vessels was prepared in 4 cadavers for simulated purpose. Clinically, two patients with tibial nonunion were treated by transplantation of this pedicled bone flap. RESULTS: In the 4 simulated operations, the ink which was injected into the femoral artery could be released from the joint branches. And in the clinical operation, the area of the pedicled bone flaps were 5.0 cm x 2.0 cm x 1.5 cm and 4.5 cm x 1.5 cm x 1.5 cm respectively. Followed up for 8 to 12 weeks, the fracture unioned well. CONCLUSION: This operation is simple and reliable, it is characterised by 1. unnecessary to anastomosis the vessels 2. reliable blood supply 3. high quality of bone flap 4. either bone flap or bone skin flap can be chosen 5. long vessel pedicle. PMID- 12563927 TI - [Experimental study of actin and myosin in the scar tissue]. AB - OBJECTIVE: To investigate the different expression of actin, myosin II in hypertrophic scars, keloids and normal skins, and to understand the relationship of actin, myosin II and the scar contracture. METHODS: Fifteen cases with hypertrophic scars, 10 cases with keloids and 15 cases with normal skins were chosen randomly. The expression of actin and myosin II were detected by immunohistochemical method in the hypertrophic scars, keloids and normal skins. The fibroblasts isolated from three types of tissue were cultured in vitro, then actin and myosin II in three different fibroblasts were measured using flow cytometry. RESULTS: The immunohistochemical staining of myosin II in hypertrophic scars was positive, while the staining in keloids and normal skins were negative. The positive rate of myosin II expression in hypertrophic scars, keloids and normal skins were (95.11 +/- 2.78)%, (16.86 +/- 7.11)%, and (5.31 +/- 1.79)% respectively. There were significant difference between keloids and the two others(P < 0.01). The actin expression in three difference tissues were positive, there were no significant difference in hypertrophic scars, keloids and normal skins(P > 0.05). The positive rate of actin expression in hypertrophic scars, keoids and normal skins were(77.77 +/- 15.43)%, (88.89 +/- 10.29)%, and (82.92 +/ 13.48)% respectively, and there were no significant difference(P > 0.05). CONCLUSION: Myosin II may play an important role in the scar contracture. Actin is the contractile protein of cell, it plays important role in cellular movement. Actin is necessary protein in the cell. PMID- 12563928 TI - [Repairing deformity of the head and face with tissue flap pedicled with the superficial temporal artery in children]. AB - OBJECTIVE: To evaluate the effect of tissue flap pedicled with the superficial temporal artery in repairing deformity of the head and face in children. METHODS: From October 1986 to December 1996, 13 children with deformity of the head and face were repaired by this tissue flap. Among them, there were congenital deformity in 9 cases, burned scar in 3 cases and infection scar in 1 case. Among the flaps, 1 was temporal skin flap, 3 were temporal flap with hairbearing scalp, 1 was frontal skin flap, and 8 were posterio-uricular superficial fascia flap and skin flap. The area of tissue flap was ranged from 5.0 cm x 1.2 cm to 10.0 cm x 5.0 cm. The length of the pedicle was 5-8 cm. RESULTS: All tissue flaps healed with first intention. Followed up for 6 months to 12 years, the appearance and function of tissue flaps were satisfactory. CONCLUSION: The tissue flap pedicled with the superficial temporal artery is suitable to repair many kinds of deformities of the head and face in children. It has the advantages of good blood supply, closely acceptor area, easy operation and satisfactory appearance. PMID- 12563929 TI - [Prophylaxis for deep vein thrombosis with low molecular weight heparin following hip and knee surgery]. AB - OBJECTIVE: To evaluate the efficacy and safety of low molecular weight heparin(LMWH) in prophylaxis of postoperative deep vein thrombosis (DVT) following hip and knee surgery. METHODS: From April 1997 to October 1998, 46 patients undergoing hip and knee orthopedic procedures were randomized into 2 groups for studying. The following eligibility criteria were applied: age over 40 years old, no recently history of venous thromboembolism (over 3 months), normal result of preoperative hemostasis test and normal result of Doppler examination of the lower extremities. One group was control group and the other group received subcutaneously a low molecular weight heparin(Fraxiparine) with anti factor X, activity of 41 IU/kg.day for three days, then 62 IU/kg.day from the 4th day to 10th day. All patients had venegraphy performed in the operated leg at 4 to 7 days after surgery. RESULTS: eight patients(34.8%) developed DVT in the control group of 23 patients and 1 patient (4.3%) in the experimental group, also of 23 patients(P < 0.05). Two groups had no any bleeding complications. CONCLUSION: The low molecular weight heparin is safe and effective in preventing postoperative deep vein thrombosis in patients following hip and knee surgery. PMID- 12563930 TI - [Mechanism study of electrothrombosis by copper needle]. AB - OBJECTIVE: To investigate the mechanism of electrothrombosis by copper needle, in order to supply the referential data for clinical treatment of vessel deformity. METHODS: The mechanism and condition of thrombus formation by copper needle were studied in vivo and in vitro using electrophysics, atom absorption spectrophtometry, histological, and histochemical methods. RESULTS: Great deal of copper ion was dissociated, and agglutination of red blood cells(RBC) in blood could be observed in vitro after the current applied by copper needles. Formation of stable thrombus was related to voltage and time of application of electric current. CONCLUSION: Dissociation of copper ion and agglutination of RBC are the basic principle of electrothrombosis with copper needle. A 4V direct current and 17.5 minutes are the safe and effective conditions for thrombus formation in the blood vessels. PMID- 12563931 TI - [Relationship between exposure time and cryopreservation of articular cartilage]. AB - OBJECTIVE: To search an optimal method for improving viability of cryopreserved articular cartilage. METHODS: Articular cartilage which was sampled from the rabbits were randomly divided into 5 groups. Fresh cartilage was group I, other groups were frozen. Before frozen, other cartilage was exposured in 10% DMSO at 4 degrees C for 30 minutes(group II), 1 hour(group III), 2 hours (group IV), 4 hours(group V), then were stored in liquid nitrogen for 1 week. Viabilities of the chondrocytes were detected by Typan-blue staining, electron transmission microscope, and determination of incorporation 3H-TdR after the temperature returned to normal. RESULTS: 1. The cells were injuried at different extent after the cartilage was frozen. In group I, survival rate of cells was 96% and incorporation of 3H-TdR was (4,953.13 +/- 583.27)%, statistic difference was significant between group I and other groups(P < 0.01). The microstructure of group I was normal while other groups all had damage of the organella, 2. Structures and functions of cells in group IV were best among frozen groups. Organella were less damaged than group II, III, V, survival rate of cells was 56% and incorporation of 3H-TdR was (1,139.88 +/- 146.39)%, statistic difference was significant between group IV and group II, III, V(P < 0.01). CONCLUSION: If cartilage are exposured in 10% DMSO at 4 degrees C for 2 hours before frozen, optimal cryopreservation can be achieved. PMID- 12563932 TI - [The primary study on rabbit bone marrow mesenchymal stem cells in constructing tissue-engineered cartilage]. AB - OBJECTIVE: To study the feasibility of constructing tissue engineered cartilage by differentiated rabbit bone marrow mesenchymal stem cells(MSC) cultured in vitro and in vivo. METHODS: The MSC were isolated from the nucleated cells fraction of autologous bone marrow by density gradient centrifuge, and then induced into chondrogenic differentiation by adding dexamethasone, transforming growth factor-beta 1 (TGF-beta 1) and ascorbic acid in vitro. After 3 weeks, some cells turned to round shape and secreted metachromatic matrix. The cartilaginoid grafts composed of chondrogenic MSC. Bovine type I collagen and human fibrin were cultured within the chondrogenic medium for 2 weeks in vitro or transplanted subcutaneously adjacent to the knee joint for 3 weeks in vivo. RESULTS: The most cells in the grafts were degenerated and disappeared after cultured in vitro. But the residual cells were survival and secreted metachromatic staining proteoglycan with toluidine blue, which was characteristic cartilage matrix. The grafts developed into matured cartilage tissue assessed by histological examination after 3 weeks of transplantation in vivo. CONCLUSION: MSC can be used as functional cells to constructing tissue engineered cartilage. PMID- 12563933 TI - [Repair of growth plate defects of rabbits with cultured cartilage transplantation]. AB - OBJECTIVE: To prevent early closure of growth plate and developmental deformities of limbs by allografts of cultured cartilages into growth plate defects of rabbits. METHODS: Chondrocytes isolated from articular cartilage of 1-month rabbits formed cartilage after cultivation in centrifuge tubes. The cartilages cultured for two weeks were implanted into growth plate defects of proximal tibiae of 6-weeks rabbits. At 4th and 16th weeks, X-ray, histologic and immunohistochemical examination were performed. RESULTS: The tibiae had no marked deformities after 4 weeks of operation. Histologic examinations showed that the defects were filled with cartilage. Immunohistochemical results of type II collagen were positive. The tibiae with allografts of cultured cartilages had no evident deformities after 16 weeks of operation. Histologic examination showed nearly closure of growth plates. On the contrary, the tibiae on control side formed severe deformities and growth plate were closed. CONCLUSION: Allograft of cultured cartilages into growth plate defects may replace lost growth plate tissues, maintain normal growth of limbs and prevent developmental deformity. PMID- 12563934 TI - [The preliminary result of osteoid callus allograft for bone healing]. AB - OBJECTIVE: To investigate the feasibility of osteoid callus allograft as a kind of bone healing promoting materials. METHODS: The osteoid callus was obtained at one week after bilateral femoral fracture of a SD rat, then was kept at -196 C for 2 weeks. The bone defect model which bone repair was in intra-membranous osteogenesis was made at bilateral tibial shaft in 5 rats, and filled with the osteoid callus in the left defect area, the right side was filled with allogenous cancellous as control group. The specimen were processed with undecalcified technique and the sections were staining with light blue and sofranin T. RESULTS: After 2 weeks, there were cartilage and bone formation in the defect area of osteoid callus graft group(3/4), medullary cavity formation in bone tissue with cartilage arround it, fibrous tissues between new bone and host bone. While there were no cartilage or bone formation in the control group. CONCLUSION: The allograft osteoid callus is not absorbed by immunological rejection, but changed into bone tissue through endochondral osteogenesis. It is inspiring to develop osteoid callus allograft as a kind of material for bone healing. PMID- 12563935 TI - [The expression of transforming growth factor alpha and its receptor in nasal polyps]. AB - OBJECTIVE: To assess the possible role of expression of TGF alpha and EGFR in nasal polyps and its relationship with PCNA labeling index. METHOD: Specimens from 20 patients of nasal polyps were studied with immunohistochemical technique. RESULT: The expression of TGF alpha, EGFR and PCNA were increased in the epithelium, gland cells and inflammatory cells of nasal polyps. There was a close correlation between the intensities of TGF alpha, EGFR and PCNA. CONCLUSION: TGF alpha may play a key role in epithelial cell proliferation in nasal polyps. PMID- 12563936 TI - [A study on biological characteristics of normal adult human nasal septal chondrocytes cultured in vitro]. AB - OBJECTIVE: To study the characteristics of normal adult human nasal chondrocytes cultured in vitro. METHOD: The adult human nasal septal chondrocytes were cultured in vitro. The changes of cellular morphology from primary to subculture were investigated; to observe the proliferation of chondrocytes by the growth curve and the survive rate of chondrocytes; and to determine the biologic synthesis of the chondrocytes by toluidine blue, alkaline phosphatase staining and type II collagen immunohistochemistry reaction. The survive rate of the nasal septal chondrocytes was observed after being frozen (-196 degrees C). RESULT: The morphology of the primary chondrocytes maintained the spherical shape, and some became polygonal shape after 4 days. Most of the chondrocytes transformed into the shape of the fibroblast cell since the fifth generation. The number of the chondrocytes (2th generation) after being cultured in vitro for 5 days was approximately 4 fold of that of the plating. The phenotype of the chondrocytes was maintained in the first 4 generation, the survive rate of the chondrocytes suscitating from being frozen (-196 degrees C) was 93%. CONCLUSION: The normal adult nasal septal chondrocytes cultured in monolayer in vitro maintained the specific chondrocytes phenotype in the first 4 generation, the growth was normal, and could survive in the condition of deep-frozen. PMID- 12563937 TI - [Study on the association between interleukin-5 and eosinophil in nasal polyp]. AB - OBJECTIVE: To explore the expression of IL-5 and its role in the formation and development of nasal polyp. METHOD: 31 patients with nasal polyp, 11 patients with chronic sinusitis(CS) and 6 control cases were chosen. Their IL-5 concentration in tissue homogenate were measured by ELISA. All patients with chronic sinusitis, control cases and 15 patients with nasal polyp were chosen to be counted eosinophils in their HE slice. RESULT: 1. The values of IL-5(pg/ml) in nasal polyp, CS and control group separately were: 23.44 +/- 6.68, 16.41 +/- 3.09, 12.86 +/- 4.17. IL-5 concentration in nasal polyp group was higher than that in the other two groups(P < 0.001). 2. The numbers of eosinophils in nasal polyp, CS and control group were 7.42 +/- 2.33, 1.30 +/- 0.59, 1.07 +/- 0.70 separately, the numbers of eosinophils in nasal polyp group were higher than that in CR or control group (P < 0.05). CONCLUSION: Nasal polyp is a disease characterized by eosinophilia, and IL-5 which activates eosinophil plays part role in the formation of nasal polyp. PMID- 12563938 TI - [The pathological changes of nasopharyngeal carcinoma cases treated by stereotactic radiosurgery]. AB - OBJECTIVE: To study the pathological changes of nasopharyngeal carcinoma cases after the treatment of stereotactic radiosurgery. METHOD: 15 cases with recurrent or residual squamous cell carcinoma of nasopharynx diagnosed as T1-4 N0M0 were selected, which had undergone previous radiotherapy. The patients were treated by Gamma Knife while the isodose curve was 50% and the margin dose was 20 Gy. The nasopharynx biopsy was performed before the treatment and 1, 3, 6, 12 months after the treatment. The biopsy specimen was taken to make a pathological examination. RESULT: 1. Before the Gamma Knife treatment, carcinoma cell could be seen in the tissue; 2. 1-3 months after the treatment, cell necrosis and acute inflammation cell infiltration could be seen in the target; 3. 6-12 months after the treatment, infiltration of chronic inflammation cell, proliferation of fibrous tissue and capillary could be found in the target. CONCLUSION: This research implies that the short-term pathological changes after the treatment of stereotactic radiosurgery can be defined as two phases: The first phase occurs from 1 to 3 months after the treatment called necrosis period. The second phase occurs from 6 to 12 months after the Gamma Knife treatment named as absorption period. PMID- 12563939 TI - [Nasal cavity volume and nasopharyngeal cavity volume in adults measured by acoustic rhinometry]. AB - OBJECTIVE: To investigate the range of normal value of total nasal cavity volume(NV) and total nasopharyngeal cavity volume (NPV) of healthy adults. METHOD: Six hundred and fifty-nine healthy adults and eighty-two adults of chronic rhinitis were measured with acoustic rhinometry. RESULT: The range of NV was 29.922-37.481 cm3 and NPV was 29.369-44.159 cm3. Comparing healthy adults with adults of chronic rhinitis, there was a significant difference in NV. CONCLUSION: Acoustic rhinometry suited for objective assessment of the nasal airways in adults. It was demonstrated that these data could provide available information for the study of nasal physiology and pathophysiology, as well as for the diagnosis and judgement of therapeutic effectiveness of nasal diseases. PMID- 12563940 TI - [Intranasal endoscopic treatment of chronic sinusitis in children]. AB - OBJECTIVE: To explore the curative effect of endoscopic sinus surgery (ESS) and the factors associated with surgery effect for treating children with chronic sinusitis. METHOD: Thirty-one children with a median age of 10 years (range 5-14 years) who suffered from chronic sinusitis or/and nasal polyps and were operated via ESS from May 1996 to January 1999 were retrospectively analyzed. RESULT: According to the therapeutic evaluation standard (ESS-1997, Haikou), twelve cases (38.7%) were completely cured, fifteen cases (48.4%) were improved and four children (12.9%) showed no change with a general effective rate of 87.1% without any severe operative complication. CONCLUSION: The results suggested ESS is a safe and effective method in the treatment of children with chronic sinusitis or/and nasal polyps. Furthermore, meticulous postoperative endoscopic care and medication are also important for securing optimal long-term results. PMID- 12563941 TI - [Bacteriological study of single nasal sinusitis and sensitivity of antibiotics]. AB - OBJECTIVE: To properly understand the bacterial distribution and susceptibility to antibiotics in the sinus of nasal sinusitis. METHOD: The mucosal and secretion in sinus were taken from 135 patients who suffered with single nasal sinusitis, by functional endoscopic sinus surgery, were cultured for bacteria. Meanwhile the antimicrobial susceptibility was determined for ordinary antibiotics. RESULT: The bacterial growth was present in 88.15% of cases, mixed bacterial growth was present in 52.10% of all bacterial, anaerobe growth was present in 39.06% of all bacteria, the susceptibility to penicillin was in 53.12% of all bacteria and cefaolin was in 87.50%. The susceptibility to metronidazole was in 94.67% of all anaerobes. CONCLUSION: We think the method that penicillin in combination with streptomycin and metronidazole, sufameth oxazole united with metronidazole, cefaolin added metronidazole were effective in nasal sinusitis treatment before the antimicrobial susceptive examination had been done. PMID- 12563942 TI - [The long-term effect of augmentation rhinoplasty with silicone]. AB - OBJECTIVE: To evaluate the long-term effect of augmentation rhinoplasty with silicone. METHOD: 360 patients underwent augmentation rhinoplasty with silicone. Improved "L" shaped implants were used in 343 patients. RESULT: Among 360 patients, 334 cases were successful enough (92.83%). CONCLUSION: The key to the maintenance of the long-term effect is to use properly improved "L"-shaped implant, to choose a suitable tunnel, to prevent various complications and to ask patient's opinions for implant design. PMID- 12563943 TI - [Treatment of patients with chronic sinusitis by endoscopic and Caldwell-Luc approaches]. AB - OBJECTIVE: To investigate the clinical effect of surgical procedures on patients with chronic sinusitis. METHOD: 104 patients with chronic sinusitis were treated by endoscopic and Caldwell-Luc's approaches. The lesions were cleared away completely. RESULT: The clinical cure rate was 96.1% after 6-12 months (average 9 months) follow up. CONCLUSION: It suggests that patients with severe maxillary sinus diseases might be cured by endoscopic and Caldwell-Luc's procedure. PMID- 12563944 TI - [A preliminary study on the incidence of cedar pollinosis in district of Wuhan]. AB - OBJECTIVE: To investigate the incidence of cedar pollinosis in district of Wuhan. METHOD: A survey relative to cedar pollinosis was carried out in 309 students in Hubei Medical University in 1999, including questionnaire investigation, nasal inspection and scratch test. From February to April of 2000, a clinical investigation of cedar pollinosis was adopted among 205 patients of allergic rhinitis including scratch test and nasal mucosa test. RESULT: 1. The positive rate of cedar pollen scratch test of 309 students was 7.8%. 2. 59 cases (28.8%) was hypersensitive to cedar pollen clinically in allergen test. CONCLUSION: Cedar pollinosis is one kind of important pollinosis in district of Wuhan and should be paid high attention to. PMID- 12563945 TI - [MDM2 gene expression in nasopharyngeal carcinoma and its relationship with p53 protein expression and EB virus latent infection]. AB - OBJECTIVE: To define the expression levels of MDM2 gene in nasopharyngeal carcinoma (NPC) and its relationship with p53 protein expression and EB virus latent infection. METHOD: MDM2 gene expression at mRNA and protein levels, p53 protein and EB virus DNA were detected by nonradioactive in situ hybridization (ISH), immunohistochemistry(IHC) and polymerase chain reaction (PCR) separately in 46 cases of NPC tissues and 12 cases of chronic inflammation of nasopharyngeal epithelium (CINE). RESULT: Fourteen cases of NPC showed MDM2 mRNA and protein overexpression, 38 cases were p53 protein positive, and 43 cases were EBV-DNA positive. Neither MDM2 nor p53 protein was expressed in any case of CINE. MDM2 expression was significantly related to p53 protein expression (P < 0.05), but not to EB virus latent infection in NPC. CONCLUSION: MDM2 gene may play an important role in the pathogenesis of NPC through interacting with p53 protein. PMID- 12563946 TI - [The relationship between ECochG and glycerol test in vertigo patients (report of 112 cases)]. AB - OBJECTIVE: For further investigating the diagnostic value of the ECochG in Meniere's disease. METHOD: 112 vertigo patients including Meniere's disease, sudden deafness, cervical vertigo and other vertigo disease, had been done ECochG, glycerol test, audiometry and caloric test. The results were compared with those of normal group. RESULT: In Meniere's disease, the positive rate of abnormal-SP/AP in ECochG was 54.69% and the positive rate of glycerol test was 53.12%. There are no statistics difference (P > 0.05) between this two tests with X analysis. CONCLUSION: We think that the glycerol test and other relative examination are necessary for the diagnosis of Meniere's disease while the-SP/AP > 0.4 ms in the examination of ECochG. PMID- 12563947 TI - [Observation and analyses on voice changes in the normal aged]. AB - OBJECTIVE: To measure the voice samples of the normal aged in order to systemically study the features of the voice changes. METHOD: To collect and analyze 146 voice samples of the normal aged with sonogram. RESULT: The fundamental frequency of the voice of the aged decreases and rises in the male more than 80 years older. The low frequency harmonics are regulation and the intensity is strong in the formant of the aged. The difference reduces in voice between male and female. The harmonics to noise ratio tends downwards and the amplitude perturbation quotient tends upwards along with the growth of age in the aged male. The changes of the above-mentioned parameters are not significant in the aged female. CONCLUSION: The voice changes are normal physiological ones in the normal aged. The changes of the parameters are used to evaluate normal aged voice and abnormal one. The changes show that the function in the aged phonation tends to decline to a certain extent and it must be protected and be trained. PMID- 12563948 TI - [p27 protein expression and its significance in human laryngeal carcinoma]. AB - OBJECTIVE: To determine clinical significance and expression of p27 protein in human laryngeal carcinoma tissue. METHOD: To measure p27 protein expression in 50 cases laryngeal carcinoma tissue by immunohistochemical LSAB (labelled streptavidin biotin) method, 15 cases vocal cord polyps used as control. RESULT: There was significant difference between p27 protein expression in laryngeal carcinoma and in vocal cord polyps, among pathological staging of laryngeal carcinoma (G1-G2 and G3), between the survived and the dead laryngeal carcinoma patients 3 years after operation, but no significant difference among clinical staging and between the patients with and without lymph node metastases. There was significant difference between the survival rate curves (Kaplan-meier curve) in p27 protein expression positive group and that in p27 protein expression negative group. CONCLUSION: p27 Protein expression is related to malignant degree of laryngeal carcinoma, so it can be an important marker of identifying the malignancy and prognosis of laryngeal carcinoma. PMID- 12563949 TI - [Immunohistological observation of tonsillae in IgA nephropathy]. AB - OBJECTIVE: To realize the relationship between IgA nephropathy (IgA-N) and immunological abnormality of tonsillae. METHOD: Thirty-one patients were diagnosed as IgA-N via renal puncture. Sixteen patients suffering from chronic tonsillitis as control. All resected tonsillae were labelled immunohistologically by ABC method. RESULT: The differences of expression of IgA, IgG in the tonsillae between above two groups were significantly obvious. The number of lymphocytes which can produce IgA increased in IgA-N tonsillae. CONCLUSION: The immune functions are abnormal in IgA-N patients. Tonsillectomy can prevent the antigen invasion, eliminate IgA originated from tonsillae, decrease the IgA immunocomplex in circulation, therefore decrease the IgA immunocomplex deposition in the basal membrane of glomeruli. PMID- 12563950 TI - [Study on the relationship between microvascular density and prognosis in laryngeal cancer]. AB - OBJECTIVE: To study the relationship between the microvascular density(MVD) and the prognosis in laryngeal cancer. METHOD: Microvessel quantitation in 31 cases of laryngeal cancer was done with factor VIII-related antibody and SABC method. RESULT: The quantitation of microvessels in laryngeal cancer were negative of histologic grade and type, but were significantly different (P < 0.05) between metastasis (10.23 +/- 0.78) and without metastasis(6.38 +/- 0.33) two years after operation. CONCLUSION: The angiogenesis of laryngeal cancer is correlated with the lymph node metastasis and prognosis. PMID- 12563951 TI - [The clinical value of laryngeal image analysis with two methods]. AB - OBJECTIVE: To analyze normal and pathological glottal image quantitatively with computed image technique, and try to calculate the absolute value of the laryngeal parameter. METHOD: The glottal image parameters of 60 normal adults, 60 cases of vocal polyps and 40 unilateral vocal fold paralysis(UVFP) patients were measured and analyzed, including the relative and absolute maximum or minimum glottal area, that is Amax(r), Amin(r), Amax(a) and Amin(a) respectively. RESULT: Amin(r) of females was larger than that of males; and both were larger than zero. He and she were equal in Amin(a) and Amax(r), while Amax(a) of males was larger than that of females. Compared with normal adults, Amin(r) of the patients with vocal polyp increased, and Amax(r) of UVFP patients diminished, while their Amin(r) increased. CONCLUSION: Laryngeal image could be analyzed quantitatively with computer assisted fibrolaryngoscopy. Normal adults have incomplete glottal closure to certain degree, and the level is different between sexes. The patients with vocal polyp and UVFP have relevant pathological glottal image. PMID- 12563952 TI - [Reconstruction of hypopharyngeal cavity with neck reverse rectangular thinned skin flap]. AB - OBJECTIVE: To study the postoperative convalescence of swallow after reconstruction of hypopharyngeal defect with the neck reverse rectangular thinned skin flap. METHOD: A thinned reverse skin flap was designed in the middle line of a platysma skin flap, and was used in hypopharyngeal reconstruction for 20 patients with squamous cell carcinoma of the hypopharynx. RESULT: No skin flap complications occurred in all patients. Fistulas in 5 cases healed spontaneously in 7 days to 10 days without further surgery. One case with lower anastomotic site stenosis was performed by subsequent successful surgery. The survival rate was 50% and 33.3% respectively after 3 years and 5 years follow up. CONCLUSION: This flap was effectively applied for reconstruction of hypopharyngeal defect for hypopharyngeal cancer, which was easy to master by surgeons, and was a high successful procedure of choice. PMID- 12563953 TI - [The role of tracheotomy in saving the patients suffered from lung blast injury]. AB - OBJECTIVE: To explore the role of tracheotomy in saving the patients suffered from lung blast injury. METHOD: To analyse the pathological change of lung blast injury and to make a retrospective analysis for clinical data of 8 cases. RESULT: Three severe injuries accompanying asphyxia, SaO2 < 75%, were tracheotomized and sucked hemorrhagic frothy sputum in trachea at once, and they were saved successfully. The other 5 cases, SaO2 > or = 90%, were not tracheotomized. CONCLUSION: During saving the lung blast injury, tracheotomy has an important role in sucking hemorrhagic frothy sputum in trachea and keeping the respiratory tract unobstructed. PMID- 12563955 TI - [Huge benign mesenchymoma in the pharynx-esophagus]. AB - OBJECTIVE: To investigate the diagnosis and management of the benign mesenchymoma in pharynx-esophagus. METHOD: Records of 2 cases with the benign mesenchymoma, arising from the pharynx-esophagus, were reviewed. RESULT: These tumors, their pedicle attached to the lateral pharynx or esophageal orifice and the body fall down to the esophagus, were large and solitary. Complete surgical resection is believed to be the most suitable treatment by lateral pharyngotomy. CONCLUSION: The benign mesenchymoma in pharynx-esophagus is a rare disease which might be misdiagnosis. Diagnosis is based on typical histologic features as well as clinical information. The prognosis is favorable. PMID- 12563954 TI - [The treatment of hypopharyngoesophageal stenosis]. AB - OBJECTIVE: To improve the curative effects of hypopharyngoesophageal stenosis. METHOD: 35 patients treated for hypopharyngoesophageal stenosis were retrospectively reviewed. The curative methods were selected on the stage of stenosis, including drug, dilatation, reconstruction of hypopharynx and oesophagus. RESULT: 33 patients resumed a normal oral diet, no stenosis of anastomosis occurred. CONCLUSION: For the early mild lesions, using of steroids, antibacterial and dilatation is considered to be the procedure of choice, for those severe lesions, reconstruction of hypopharynx and oesophagus with selected appropriate materials can obtain good results. PMID- 12563956 TI - [Effects of modified SD therapy on Bell's palsy]. AB - OBJECTIVE: To investigate the effects of a modified SD therapy (intravenous high dose hydrocortisone and low molecular dextran) on Bell's palsy(BP), and to avoid the severe side-effects such as hepatic and renal disorders during the treatment. METHOD: Seventy-one BP patients were treated with modified SD method (modified group), thirty-two BP patients took prednisone (control group), and the curative rates of two therapies were compared, in the meanwhile recorded the side effects in detail. RESULT: Curative rate in modified group was 95.8% (House-Brakmsnn I II) and in control group was 81.2% (P < 0.05). In modified group, the curative rate (I grade) in the patients were treated within 24 hours following onset was 75.0%, and the curative rate in other three subgroups were treated within 24-48 hours, 2-3 day and 3-5 day following onset was 43.8%, 26.7%, 18.8% respectively. Modified group had not shown hepatic and renal disorders or gastric ulcer. CONCLUSION: Modified SD method may increase recoverable rate and can avoid significant side effects for BP, and emphasizes the importance of the administration of SD therapy in the early stage of the disease. PMID- 12563957 TI - [Treatment of non-invasive aspergillosis of maxillary sinus by functional endoscopic sinus surgery]. AB - OBJECTIVE: To evaluate the treatment of non-invasive aspergillosis of the maxillary sinus by using functional endoscopic sinus surgery (FESS) and the correlated factors that affect the treatment. METHOD: 41 cases of local, non invasive aspergillosis of the maxillary sinus undergone FESS were studied. RESULT: The period of convalescence of double pathway (via ostium of maxillary sinus and canine fossa) was 4.7 weeks while single pathway (via ostium of maxillary sinus only) was 9.3 weeks. There was significant difference (P < 0.01). The period of convalescence of group using anti-fungal drugs to wash the operative cavity was 6.4 weeks while the group without using drugs was 6.7 weeks. There was no statistic significance (P > 0.05). CONCLUSION: FESS is effective in treating of non-invasive aspergillosis of the maxillary sinus. The operation of double pathway is superior to that of single pathway. Whether to use antifungal drugs to wash the operative cavity has no obvious effect on the efficacy. PMID- 12563958 TI - [Clinical analysis of facial nerve palsy in middle ear and mastoid surgery in 23 cases]. AB - OBJECTIVE: To investigate the cause, the prevention and the therapy of facial nerve palsy(FNP) which induced by the operation of middle ear and mastoid surgery. METHOD: A series of 1032 cases undergone middle ear and mastoid surgery were reviewed between 1976 and 1991. RESULT: 23 cases FNP were identified. All were incomplete and peripheral. 13 cases of them were performed decompression of facial nerve. 6 cases were cure, 4 cases on the mend and 3 cases no effect after operation. 10 cases were treated by conservation, 5 cases were cure, 3 cases on the mend and 2 cases no effect. CONCLUSION: If the facial paralysis occurs during the operation, which means that facial nerve is damaged, except hard-stuff reason. Exploration should be done at once. If facial paralysis occurs later, conservative therapy should be selected at first. If facial paralysis is still not on the mend 1 month of postoperation, one should be done exploration of facial nerve. PMID- 12563959 TI - [Experimental study on repair of cricoid cartilage defect with cartilage grafts in rabbits]. AB - OBJECTIVE: To investigate the effect of free cartilage fragment in repair of cricoid cartilage defect. METHOD: One-third of cricoid cartilage was cut and repaired with free thyroid cartilage fragment and pedicled cartilage in rabbits. The pathological changes of the cartilages were observed. RESULT: 1. The free thyroid cartilage fragments were absorbed in two months, and were replaced by scars. 2. The pedicled cartilage fragments were not absorbed and healed with the cricoid cartilage. 3. No evidence of growth impairment of surgery-induced cricoid cartilage was observed. CONCLUSION: In rabbits, both free cartilage fragments and pedicled cartilage fragments could repair the defect of cricoid cartilage effectively. PMID- 12563960 TI - [Comparison of the effectiveness of levofloxacin and cefuroxime for the treatment of sinusitis]. AB - OBJECTIVE: To observe the effectiveness of levofloxacin and cefuroxime for the treatment of sinusitis in adults. METHOD: Patients with acute or chronic sinusitis were randomly assigned to receive either levofloxacin (200 mg orally once daily, 200/100 mg twice daily) or cefuroxime (250 mg orally twice daily) for 10 to 14 days. Pre- or post-treatment, the clinical and laboratory examination were done and in the end of treatment, the efficacy and safety were assessment. RESULT: The success rates after the end of treatment were 97.4% for the 76 patients who received levofloxacin and 92.8% for the 14 patients who received cefuroxime. The resolution rates of bacteria were 91.6% and 80.0%, respectively. The safety of these two groups were more high. CONCLUSION: Levofloxacin is effective for the treatment of sinusitis in adults and pay attention to the course of treatment for raising the efficacy of treatment. PMID- 12563961 TI - [Selective neck dissection in the treatment of supraglottic carcinoma]. AB - OBJECTIVE: To study the effectiveness of selective neck dissection (upper neck dissection, UND) in the treatment of N0 neck of supraglottic carcinoma. METHOD: The upper neck dissection was designed according to the lymphatic drainage of the supraglottic larynx for the management of N0 neck. The records of the supraglottic carcinoma of T1-4 N0-1 M0 cases (168 patients), who were treated in 1976-1990 at this hospital, were analysed retrospectively. RESULT: In this seires of 168 patients with no neck metastases pathologically a follow-up of five years after an upper neck dissection revealed a neck recurrence rate of 10.1% (17/168), which is comparable with those reported in the literature after selective neck dissection. Five year survival rate was 72.6% (122/168). CONCLUSION: Long-termed observation after UND for supraglottic carcinoma (T1-4 N0-1) resulted in satisfactory survival rates and regional control. It seems justifiable to do a selective neck dissection for N0 and selected N1 patients in order that more patients were exempted from enduring the morbidity following a comprehensive neck dissection. PMID- 12563962 TI - [The influences of vowel ae and a: on acoustic measures]. AB - OBJECTIVE: To study the influences of vowel ae and a: on acoustics measures. METHOD: Fifty-three cases of laryngeal diseases were measured acoustically. The acoustic sampling were vowels ae and a:. RESULT: The results showed that Fo, Jitter and Shimmer were similar between two groups, and NNE of vowel a: was much lower than that of ae, indicating larger glottal closure in vowel a:. CONCLUSION: Vowels have something to do with glottal closure, therefore influencs acoustical measures. PMID- 12563963 TI - [Clinic analysis of surgical treatment for benign parotid tumour 55 cases]. AB - OBJECTIVE: To investigate the relationship between 3 kinds of surgical methods and the postoperative recurrence and complication of patients with the benign parotid tumours. METHOD: The records of benign parotid tumours cases (55 patients), who were treated in 1987-1997 at our hospital, were retrospectively analysed. The operative methods as follows: local excision (20 patients), superficial parotidectomy (28 patients), total parotidectomy (7 patients), facial nerve was anatomied during operation for 35 patients. RESULT: The rate of tumour recurrence, partial facial nerve paralysis, parotid gland fistula and Frey's syndroms after operation were 14.6% (8/55), 14.6% (8/55), 7.3% (4/55) and 7.3% (4/55) respectively. CONCLUSION: The operation of superficial parotidectomy or total parotidectomy should be perfromed for benign parotid tumour cases, facial nerve should be anatomied during operation, for reducing tumour recurrence and facial nerve paralysis. PMID- 12563964 TI - [Surgical treatment of laryngeal carcinoma in the elderly over 70]. AB - OBJECTIVE: To explore the possibility and perioperative management of laryngectomy in the elderly over 70. METHOD: The records of 28 patients of laryngeal carcinoma aged 70 and over treated with laryngectomy were reviewed. RESULT: Among the 28 cases, 23 cases (82.1%) had one or more concomitant diseases or laboratory abnormalities. Most of them were cardiovascular disorders (60.7%) and respiratory disorders (25%). Total laryngectomy was performed in 15 cases. Partial laryngectomy was performed in 12 cases. Vocal cardectomy was performed in 1 case. The neck dissection was performed in 3 cases. No operative accident and death had happened in this group. The rate of postoperative complications was 32.1%. CONCLUSION: The elderly over 70 can tolerate laryngeal surgery well if the case is properly selected and careful perioperative management. PMID- 12563965 TI - [Clinical analysis of non-Hodgkin's lymphoma in head-facial]. AB - OBJECTIVE: To explore the clinical features, diagnosis and treatment of non Hodgkin's lymphoma in head facial. METHOD: The clinical data of 32 cases non Hodgkin's lymphoma in head facial were analysed retrospetively. RESULT: The rate of diagnosis was low in early. It frequently misdiagnosed as chronic inflammation, malignant granulomatosis, epithelial cancer, and undifferentiated cancer, etc. CONCLUSION: The biopsy shoud be performed routinely and it could do repeatly. It is necessary to do immunohistochemical examination. Only by earlier diagnosis, prompt and reasonable treatment, the curative effect would be better. PMID- 12563966 TI - [Clinical significance and changes of fibrillation potential amplitude following denervation of laryngeal muscles]. AB - OBJECTIVE: To evaluate the value of clinical application of examination of fibrillation potential amplitude of laryngeal electromyogram. METHOD: Fifty-four patients (65 sides) with recurrent laryngeal nerve paralysis were examined and only the maximum fibrillation potential amplitudes were recorded in the posterior cricoarytenoid muscles and thyroarytenoid muscles. RESULT: There was no significant difference between sexes, ages and sides. The fibrillation potential amplitude was maximum at 3 to 4 months after denervation of the posterior cricoarytenoid muscle or at 5 to 6 months after denervation of the thyroarytenoid muscle and still remained at certain level for years in some patients. Significant difference was showed in complete and incomplete nerve injuries groups, but changes of amplitude with time in incomplete nerve injuries group were small. CONCLUSION: The changes of fibrillation potential amplitude might indicate that the changes from muscle denervation was still reversible and might be more accurate than traditional method of examination. PMID- 12563967 TI - [The correlation analysis of obstructive sleep apnea syndrome and the size of oropharyngeal cavity]. AB - OBJECTIVE: To explore the relationship between obstructive sleep apnea syndrome (OSAS) and the size of oropharyngeal cavity. METHOD: The distance from uvula basis to tongue, the distance between anterior and posterior tonsillar pillars and the uvula area were measured for 30 OSAS patients and 30 healthy controls. RESULT: The study data indicated that there was very significant difference between patients and healthy controls (P < 0.001). The results of correlation analysis indicated that there was a nagetive correlation between the distance from uvula basis to tongue, the distance from anterior to between posterior tonsillar pillars and apnea hypopnea index (AHI) (P < 0.05), but there was a positive correlation between uvula area and AHI (P < 0.01). The patient's height, weight were measured, the body mass index was positive correlation with AHI (P < 0.001). CONCLUSION: These observations suggest that the obesity, smaller pharyngerl area can play an important role in the pathogenesis of OSAS. PMID- 12563968 TI - [The histopathological observation and the clinical significance of invasion of pyriform sinus apex in pyriform sinus carcinoma]. AB - OBJECTIVE: To approach the clinical significance of pyriform sinus carcinoma invading to pyriform sinus apex. METHOD: Forty-three cases of surgical specimens from total laryngectomy and partial hypopharyngectomy of pyriform sinus carcinoma were made with the organ subserial sectioning and pathological study. RESULT: Pyriform sinus apex is the inferior 1/3 region of pyriform sinus. If pyriform sinus carcinoma invades its apex, the cancerous tissue easely infiltrated the adjacent structure and organs. CONCLUSION: The invasion of pyriform sinus apex is the beginning position and the most important sign of invading downward the tissue and surrounding organs in the inferior position of hypopharynx. When surgery treatment is performed, the total laryngectomy and hypopharyngectomy of ill hypopharynx is appropriate and necessary. PMID- 12563969 TI - [Clinical observation of curative effect of neartotal larynegectomy for advance laryngeal cancer]. AB - OBJECTIVE: To explore a method how to preserve the speech function affer larynegectomy for advance laryngeal cancer and to improve the patients' life quality. METHOD: 17 cases with advance laryngeal cancer (T3 11 cases, T4 6 cases, glottic region 8 cases, supraglottic region 5 cases, pyriform recess 4 cases) were treated with neartotal larynegectomy (Pearson operation) in our department from February 1990 to July 1995. In the operations the remained monolateral arytenoid cartilage and an about 1.5 cm-wide mucomembranous flap, which joined to trachea, reconstructed vocal duct, lymphoglandulae near the branch of neck artery were checked routinely in those N0 cases, and the radical neck dissection was taken or not according the result of frozen section. RESULT: After the operation good speech function was obtained in 12 cases. All patients, except one, had good swallowing. Survival rate of 2, 3 and 5 year was 70.6%, 64.2% and 50% respectively. CONCLUSION: It is key that neck dissection should be performed rightly and the principle of surgery should be carried out for successful Pearson operation. PMID- 12563970 TI - [The objective evaluation of hearing aids]. AB - OBJECTIVE: To objectively evaluate the effect of hearing aids. METHOD: The procedure involved eliciting ABRs by clicks and slow vertex response (SVRs) by tone pips to the aided and unaided ear presented in sound field. RESULT AND CONCLUSION: There has high relativity between the thresholds of SVRs and free field pure tone audiometry. There has no significant difference between the gains from SVRs and free field pure tone audiometry. PMID- 12563971 TI - [Analysis for the auditory brainstem responses of guinea pigs with wavelet transform]. AB - OBJECTIVE: To fine out the advantages of wavelet transform when it is applied in analysis of auditory brainstem responses. METHOD: Thirty of male, albino guinea pigs weighing 300 to 400 g were divided into three groups: 1. normal hearing; 2. conductive hearing loss; 3. sensorineural hearing loss. ABR of the animals was collected and represented with wavelet transform respectively. The signals before and after being transferred was contrasted and dealt with statistics. RESULT: The ABR signals are distorted in wavelet transform scale 2 and background noises are increased in scale 4. The conventional time feature of ABR signals can be conserved and the present of I wave can be increased in wavelet transform scale 3. CONCLUSION: Wavelet transform scale 3 is proper to analyze the ABR signals of guinea pig. Compared with conventional method, it can conserve the ABR signal's time feature and increase the emerged rate of I wave. PMID- 12563972 TI - [Nerve electrogram study on different degrees injuries of recurrent laryngeal nerve]. AB - OBJECTIVE: To study the display of different degrees injuries of recurrent laryngeal nerve in electroneurogram. METHOD: We imitated several common types of injuries i.e. stretch, claming, electrocautery and cut in cervial segment of recurrent laryngeal nerve and measured latency and amplitude of the nerve by electroneurography. RESULT: The normal value of latency was (1.58 +/- 0.14) ms and that of amplitude was (2.53 +/- 0.78) mV; stretch group was (2.43 +/- 0.32) ms and (0.95 +/- 0.23) mV; claming group was (2.99 +/- 0.55) ms and (0.62 +/- 0.22) mV; electrocautery group was (4.85 +/- 1.03) ms and (0.24 +/- 0.15) mV; cut group was infinity ms and 0 mV. CONCLUSION: Latency correlated directly with the degree of nerve injury. Whereas amplitude correlated inversely with the degree of nerve injury. Stretch injured recurrent laryngeal nerve the least, followed by claming, electrocautery and cut. This method could be used for nerve monitoring during operation and prognostic judgement. PMID- 12563973 TI - [Surgical reconstruction of laryngeal and tracheal stenosis]. AB - OBJECTIVE: Reconstruction of laryngotracheal stenosis. METHOD: According to the pathologic condition of the larynx and trachea, various surgical procedures were designed for the treatment of laryngotracheal stenosis. These techniques included endoscopic management, laser therapy and laryngotrachoetomy with autogenous tissue and stent. RESULT: Among 36 patients, 32 were decannulated. The success rate was 88.8%. CONCLUSION: In order to improve the curation effects of laryngjotrachal stenosis, the choice of surgical procedure must be individualized to the pathologic condition of larynx and trachea, as well as the experience of the surgeon. Learning previous successful experience is also very important for a good result. PMID- 12563974 TI - [The clinical characters of myopatic pharyngoparalysis and laryngoparalysis with 3 cases report]. AB - OBJECTIVE: To improve the level of diagnosis and treatment about myopathic pharyngoparalysis and laryngoparalysis (MPL). METHOD: 3 cases of MPL were diagnosed by clinical inaterials, electromyogram, serokinase and muscle biopsy. The clinical characters are dysphagia, phonasthenia, abnormal electromyogram and high CPK et al. They were all treated with glucocorticoid. RESULT: The effect of treatment on all 3 cases was good. CONCLUSION: It is important to distinguish MPL with neuropathic pharyngoparalysis and laryngoparalysis. An appropriate glucocorticoid treatment is also impontant. PMID- 12563975 TI - [Quantitative analysis of vocal fold vibration]. AB - OBJECTIVE: To measure vocal fold vibration quantitatively and study the effects of gender and phonation mode. METHOD: The authors adopted a new computerized image analysis technique and EGG in 60 normal speakers, and the parameters of vocal fold vibration were measured and analyzed according to the loudness, pitch and gender. RESULT: With frequency increasing, CQ got smaller and OQ larger which expressed longer open phase. As intensity increased, the duration of open phase became shorter and the closing phase got relatively much shorter. Males had a longer closed phase than females. The closing duration appeared relatively shorter than opening duration in phonation. CONCLUSION: The quantitative measures of vocal fold vibration become feasible with modern computers and commercially available image analysis software. Certain features of vocal fold vibration vary with intensity variations, while others vary with gender or frequency variation. PMID- 12563976 TI - [Determination of T lymphocyte subsets in peripheral blood lymphocytes of patients with chronic tonsillitis]. AB - OBJECTIVE: To observe the cellular immunologic status of patients with chronic tonsillis. METHOD: T lymphocyte subsets in peripheral blood lymphocytes of 60 cases with chronic tonsillis were detected. RESULT: It was found that CD4, CD4/CD8 in chronic tonsillis patients decreased than that in normal control group with statistical significance (P < 0.01). CONCLUSION: This study indicates that the cellular immunologic status in patients with chronic tonsillitis was decreased. PMID- 12563977 TI - [One-stage reconstruction with the pedicle skin flap of forehead for defective tissue of head-neck tumors dissection]. AB - OBJECTIVE: To discuss the feasibility on reconstructing the defective tissue with the pedicle skin flap of forhead after head-neck tumors dissection. METHOD: 14 patients with advanced head-neck tumors were studied, whose defective tissues were reconstructed by their forehead skin flaps. The flap area ranged from 5 cm x 11 cm to 7 cm x 16 cm. RESULT: 2 cases of them were partial necrosis, 3 years and 5 years survival rates were 76.9% and 63.6% respectively. The function of breath through nose or mouth and the function of swallow were satisfactory. CONCLUSION: The forehead skin flap should be a good selection for the reconstruction of defective tissue after basicranial and nasalfacial dissection. PMID- 12563978 TI - [79 cases: a clinical analysis of hoarseness in children]. AB - OBJECTIVE: To research the clinical characters of the hoarseness in children. METHOD: 79 cases are analysed retrospectively. RESULT AND CONCLUSION: Age for this disease is usually 3 years old. People with irritable character are more easy to get it. Vocal nodule is the main cause to chronic hoarseness in children. Conservative treatment is important. PMID- 12563979 TI - [The choice of the stapes surgical style in treatments of the otosclerosis]. AB - OBJECTIVE: To investigate the effects of the stapes surgery on treatment of the otosclerosis to find out the more suitable surgical style. METHOD: 71 cases including 31 cases with stapes lifting surgery, 34 cases with partial stapcdectomy, 6 cases with total stapedectomy were summed. RESULT: Both the stapes lifting surgery and partial stapedectomy have steady effects on hearing level for a short or long time. CONCLUSION: The stapes lifting surgery should be adopted as the first surgical style for it is easier and the vestibular reaction is slighter. It will not affect the following surgical process even the two crura are broken. PMID- 12563980 TI - [The effect of partial middle turbinectomy upon the frontal sinus]. AB - OBJECTIVE: To investigate the effect of partial middle turbinectomy under endoscopy upon the frontal sinus. METHOD: A retrospective analysis of 87 patients undergone endoscopic sinus surgery was made. RESULT: Among 55 sinusitis patients (no frontal sinus) suffered Fess, 29 cases had partial middle turbinectomy and 7 cases were found postoperative frontal sinusitis. In other 26 cases without partial middle turbinectomy, 5 cases were found frontal sinusitis. No significant difference between two groups was found (P > 0.05). In 32 nonsinutis patients no postoperative frontal sisusitis was found. CONCLUSION: The partial middle turbinectomy has no effect on the incidence of frontal sinusitis. PMID- 12563981 TI - [The meaning of the expression of PCNA and c-erbB-2 in the hyperplasia of laryngeal epithelium]. AB - OBJECTIVE: This study was performed to examine the relationship between PCNA and c-erbB-2 expression and histological diagnosis. METHOD: The percentages of PCNA positive nuclei (PCNA index) were evaluated in biopsies of 10 with epithelial simple hyperplasia, 26 with dysplasia and 30 with squamous carcinoma by immunohistochemical staining. RESULT: The PCNA indices were 6.64%, 29.36% and 62.67% respectively. There were significant statistic correlation between these indices and their histological gradies of lesions (P < 0.01). C-erbB-2 was not positive in all of dysplastic lesion and weak in carcinoma (20%). CONCLUSION: The PCNA activity in dysplastic lesions of laryngeal premalignancis is higher than that in simple hyperplastic lesions but lower than that in carcinoma. The severe dysplasia with high PCNA indices should be treated clinically as the early invasive carcinom. The method should be regarded as a useful adjunct to traditional histological techniques with allowing more objective grading of epithelial hyperplastic lesions of larynx. There was no relationship between c erbB-2 expression and histolotical diagnosis of hyperplastic lesions. PMID- 12563982 TI - [Study on the effects of taxol on Hep-2 cell lines in larynx carcinoma]. AB - OBJECTIVE: To investigate the sensitivity of Hep-2 cell lines in larynx squamous cell carcinoma to the taxol in vitro and to provide experimental basis for clinical therapy. METHOD: Techniques of cell culture and randomized blank contrast design were used. The degree and dose-dependency of taxol which induced growth inhibition in Hep-2 cell lines were observed and the alteration of cell cycle was analysed by flow cytomethy. RESULT: All cells died at the concentration of taxol 1 x 10(-8) M; growth inhibition was observed clearly at the concentration of 5 x 10(-9) M and 2.5 x 10(-9) M. Flow cytometric data showed a G0/G1 arrest and apoptosis (28.96%) in two cell lines after exposure to taxol for 48 h at low and medium concentrations. CONCLUSION: There is a high sensitivity of Hep-2 cell to taxol in vitro. The cells show G0/G1 arrest and apoptosis. This results may provide reliable guidance for treating largnx carcinoma. PMID- 12563984 TI - [Noso-orbital related disease: an analysis of 39 cases]. AB - OBJECTIVE: To enhance the Diagnostic and therapeutic levels of naso-orbital diseases. METHOD: Thirty-nine cases of naso-orbital diseases were analyzed retrospectively. RESULT: Among the 39 cases, 7 had no nasal symptoms. 9 had their symptoms mainly confined to eyes and were initially consulted with ophtalmologists. 1 case was consulted with neurologist at first. The main pathologies were as following: bony absorption of orbit wall in inflammatory disorders; compression, displacement and bony absorption of orbit wall in benign tumors and mucoceles; bony destruction in malignant tumors. Tranmatic fractures might involve multiple bony structures. CONCLUSION: Nosal and orbital organs are closely related anatomically. Imaging investigations are important in diagnosis of naso-orbital related diseases. CT is helpful for early diagnosis. Treatment for benign tumors is mainly by operation, while malignant tumors should be treated by operation and radiotherapy together. PMID- 12563983 TI - [Study on the correlation between the cyclin D1 and laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To study the correlation between the expression level of Cyclin D1 and the laryngeal squamous cell carcinoma (LSCC). METHOD: 35 LSCC cases testified pathologically were included in the study. The samples obtained from each tumor tissue and adjacent normal mucosa were detected to see the expression level of Cyclin D1 with the method of RT-PCR and light density half quantive analysis. RESULT: The expression level of Cyclin D1 was significant higher in almost all turmor tissues than in the adjacent normal mucosae (P < 0.05). Overexpressed carcinomas were more frequently associated with lymph node metastases (P < 0.05). A significant association between the expression level of Cyclin D1 and TNM was found. The higher T stage, the higher the expression level of Cyclin D1 (P < 0.01). The expression level of low histological grade carcinomas was lower than that of the high histological grade carcinomas, but the difference had no statistical significance. No association between overexpression of Cyclin D1 and sex, age was found too. CONCLUSION: The result indicates that the expression level of Cyclin D1 in LSCC may be a useful marker of prognostic significance and it provide a molecular biology basis for treating the LSCC patients. PMID- 12563985 TI - [An experimental study about tiny traumatic measurement of the facial nerve fluid pressure]. AB - OBJECTIVE: To study endoneurial fluid pressure (EFP) in facial nerve of normal state. METHOD: EFP was recorded by an active, servo-nulling pressure system in 24 normal guinea pigs. RESULT: The results showed that EFP of the facial nerve was 2.87 +/- 0.51 mmHg and 2.70 +/- 0.43 mmHg in the tympanic and mastoid segment respectively. EFP in the tympanic part was similar to that of mastoid segment. In addition, EFP of facial nerve revealed rhythmic fluctuation within limits. CONCLUSION: It was suggested that measurement of facial nerve pressure by a servonulling system should be useful in evaluating the pathogenesis undelying facial palsy. PMID- 12563986 TI - [Report of myringoplasty with interlay method in 53 ears perforation of tympani]. AB - OBJECTIVE: To probe into an eutherapeutic method for the healing tympanic membrane aperture. METHOD: Fifty-three cases of tympamic membrane aperture were treated with myringoplasty of interlay method. The results were compared with that of underlay method of 168 ears in same period. RESULT: The rate of closure of perforation of tympani was 96.2% in interlay method group and 85.7% in underlay method. The hearing recovery of interlay method group was better than underlay method group. Function of Eustachian tube kept good condition in interlay method group. There was no blunting of anterior tympanomeatal angle in the interlay group. CONCLUSION: The interlay method of myringoplasty is an ideal method for healing perforation of tympani. PMID- 12563987 TI - [Observation of curative effect of radical mastoidectomy with rebuilding back wall of external and defect tympanoplasty]. AB - OBJECTIVE: To manage the back wall defect of external acoustic canal and opened mastoid cavity which was remained from radical mastoidectomy. METHOD: 31 ears with cholesteatomatous otitis media and 3 ears with skeletal ulcer were treated by primary operation which consisted of radical mastoidectomy with rebuilding back wall of external canal and tympanoplasty. RESULT: All cases were followed up for 5 years. And all of external anditory canal are normal in appearance. Sixteen ears reached auditory level for social interaction and another 16 for functional audition. CONCLUSION: The Primary operation that we mentioned above may resolve some problems such as mastoid cavity or big external canal which were remained from radical mastoidectomy. It is effective to improve hearing. PMID- 12563988 TI - [The influence of short-term pure tone exposure on amplitude of distortion product otoacoustic emissions and spectral history]. AB - OBJECTIVE: To observe the changs of the amplitude and the spectral history of DPOAE in the slightly damaged cochlea. METHOD: Fifteen of young person with normal hearing were exposed to 4 kHz short-term pure tone (Level = 130 dB SPL) for 3 minutes to analysis the change of DPOAE in details after Temporary Threshold Shift (TTS). RESULT: The comparison of pre- and postexposure showed that the thresholds of sound reflex is 9.78 +/- 6.98, 8.04 +/- 5.58, 13.0 +/- 5.58 dB HL and 8.82 +/- 6.23, 5.75 +/- 4.42, 9.05 +/- 2.27 dB HL in 0.5, 1, 2 kHz, respectively. The amplitude of DPOAE decreased 4.28 dB SPL in 6 kHz, and the spectral history rippled significantly. The coefficience of variance is 5.18 +/- 1.63%. The combination of the amplitude and the coefficience of variance indicated that 80.0% of those is abnormal. CONCLUSION: The results confirmed that the spectral history of DPOAE is a sensible tool to assess the slightly cochlear lesion together with the amplitude of DPOAE. PMID- 12563989 TI - [Effect of decompression of blood vessel on vascular compressive tinnitus]. AB - OBJECTIVE: To evaluate the diagnosis and surgical indications of vascular compressive tinnitus. METHOD: We searched carefully the possible focus causing vascular compressive tinnitus around VIII cranial nerve in cerebellopontine angle via retrosigmoid sinus approach under microscope in 40 cases. The responsible blood vessels and synechia focus was separated and one piece of auto musculomembrane was put between blood vessels and the nerve. RESULT: We had 38 cases of operation. All of them had artery compression on the surface of VIII cranial nerve, 6 of them had arachnoid synechia. After operation, the symptoms of tinnitus disappeared thoroughly. Of them 4 cases with vertigo recovered also. Recurrences were found in 2 cases in the follow-up survey which last 6 monthes to 7 years and 2 monthes. CONCLUSION: Surgical treatment of neurovascular decompression for vascular compressive tinnitus via retrosigmoid sinus approach has positive therapeutic effect as long as the diagnosis is correct. PMID- 12563990 TI - [Prevention of keloids of the earlobes]. AB - OBJECTIVE: To investigate the causing of keloids of the earlobes. METHOD: We have treated 41 keloides of earlobes in 28 cases, and analyzed the factors. RESULT: It has been found that infections, inappropriate supporter, allergy to nickel and gold, local delayed allergy are the chief factors to form keloides of the earlobes. CONCLUSION: Some measures to prevent those cousing mentioned above have been proposed. PMID- 12563991 TI - [TCD functional test for vertigo induced by ischemic cerebrovascular disorders]. AB - OBJECTIVE: To diagnose differentially the vetigo induced by some ischemic cerebrovascular disorder. METHOD: Patients with vertebrobasilar artery transient ischemic vertigo (group A), migraine (group B), hyperventilation syndrome (group C), hypertension (group D) are measured by using TCD functional examination which included blood peak velocity of systolic (Vs) and diastolic (Vd) end-period of vertibrobasilar artery of routine TCD (TCD-R), one minute hyperventilation TCD (TCD-HV) and one minute voluntary apnea TCD (TCD-B) respectively. RESULT: It showed that the Vs, Vd are decreased under the three conditions in A, B and D groups. The most apparent decrease are obversed in D group. The values of the decrease are similar between group A and B. No changes are found in C group. The abnormal Vs incidences of TCD-B measurement in group A are higher than those in group B and C, but significant lower than those in group D; and in TCD-HV test lower than group D and C, higher than group B; in TCD-R test, lower than group D, and no difference with group B and C. The abnormal incidences of Vd in group A are lower than group D and higher than group B in TCD-B test. In TCD-HV test, the group A abnormol incidences are lower than group D but higher than group B and C. In TCD-R test, the abnormal incidences are lower than group D and no difference between group B and C. CONCLUSION: The TCD measuremen is useful for differential diagnosis of vertigo induced by ischemic cerebrovascular disorders. PMID- 12563992 TI - [Histopathologically observational study of cartilage-mucosa on the eustachian tube isthmus of temporal bone with otitis media]. AB - OBJECTIVE: To explore the influence of inflammatory pathology change of otitis media upon cartilage-mucosa at the eustachian tube isthmus (ETI). METHOD: The cartilage-mucosa at ETI and bone-mucosa on the tympanic promontory of 32 temporal bones with otitis media were observated histopathologiclly in comparison with 50 normal temporal bones under light microscopy. RESULT: The eustachian tube isthmus in all the 32 temporal bone with otitis media and in 50 normal temporal bone were unblocked. The thick-measures and pathology observations of the cartilage-mucosa on ETI in the temporal bones with otitis media revealed there was no obviously inflammatory change and no significantly difference in comparison with the normal control group. But there was obviously inflammatory pathologic change on the bone mucosa of tympanic promomtory of temporal bone with otitis media. The demarcation line of completely different inflammatory reaction in middle ear cleft was exactly at the connective line between cartilage and bone of eustachian tube. CONCLUSION: With the specifically histologic structure, the cartilage-mucosa on eustachian tube is possessed of very strong function of natural defence for inflammatory desease, and it almost is impossible to lead to obstruction of eustachian tube by inflammatory influence upon the cartilage-mucosa. PMID- 12563993 TI - [Toxic shock syndrome related to the operation of nose]. AB - OBJECTIVE: To study the clinical features of Toxic shock syndrome. METHOD: The clinical information of two cases was analysed. RESULT: It indicates that this disease is caused by TSST-1 which is created by staphylococcus-aureus. The clinical features are acute toxic-shock syndrome with many complications and high mortality. CONCLUSION: More attention should be payed for it. PMID- 12563994 TI - [The examination of SIgE in perennial allergic rhinitis with mast allergy test system]. AB - OBJECTIVE: To explore the relation between serum specificity immunoglobulin E (SIgE) and skin test (intracutaneous test) as well as nasal provocation in Perennial allergy rhinitis (PAR) by Mast Allergy Test System. METHOD: Serum allergen-specific IgE (SIgE) were examined by Mast Allergy Test System in 57 case of PAR patients. RESULT: The SIgE positive rate of fannae and pteronvssinus are highest (52.6%), the sensitive rate of intracutaneous test is 88.4% and nasal provocation is 88.9%. The SIgE positive rates of cludosporium, penicillum and altemana are 26.3%, 24.6% and 21.0%, respectively. The SIgE positive rates of fannae, cockroach, shrimp, crab, egg and milk is direct proportion with intracutaneous test (P < 0.01). Pteronvssiaus and pigweek mix have no relationship with intracutaneous test (P > 0.05). In addition, the positive rate of ragweed mix SIgE is 22.8%, but there is non of this plant in South China. It is worthy of paying attention in clinic. CONCLUSION: It indicate that Mast Allergy Test System is one of the better examination for PAR allergens. PMID- 12563995 TI - [Rhinitis and maxillary sinuitis caseosa]. AB - OBJECTIVE: To investigate the effect of surgical procedures on patients with rhinitis caseosa and maxillary sinusitis caseosa. METHOD: The clinical analysis for 21 cases (24 sides) were reported in this paper, there were 6 rhinitis caseosa, 13 maxillary sinusitis caseosa, 5 rhinitis and sinusitis caseosa, all patients had caseation material in the nasal cavity or sinus cavity. All patients were treated successfully by surgery, transmaxillary sinus microsurgery and the middle meatal antrostomy were performed for 12 cases. RESULT: 21 cases who were followed-up 3-5 years after surgery were disappearance for symptoms, and did not recurring. CONCLUSION: The key of surgery is to allow free drainage for sinus ostium. PMID- 12563996 TI - [Research of experimental fistula stimuli vestibule evoked response]. AB - OBJECTIVE: To investiqate a way of stimulation for experimental vestibul evoked response. The artificial fistula of right upper semicircular canal was made in 12 guineapig. METHOD: Using a fistula stimuli probe device designed specially. RESULT: We obtain a potential consisting of 3-4 positive waveforms. We called it the fistula stimuli vestibule evoked response (FsVeR). They had a coincide character. The response disappeared when the animal died. The cervical spinal section, orbital removal and facial nerve removal had little effects to FsVeR. When severe inner ear damage produced by Gentamycin injection, the potential disappeared. The potential disappeared but ABR were existed. When the upper semicircular canal and its nerve damage. The noise screen had a little affection to FsVeR. CONCLUSION: These results indicated that FsVeR was mainly originated from the vestivbule system. The development of this recording technique provides a new method and maybe has important value for vestibule experimental research work. PMID- 12563997 TI - [Study on c-myc mRNA and its product expression during hair cell regeneration in the chick basilar papilla]. AB - OBJECTIVE: To study the effect of c-myc gene expression during hair cell (HC) regeneration in the chick basilar papilla (BP) following getamicin ototoxicity. METHOD: c-myc mRNA and its product (c-myc protein) expression in the damaged region was histologicolly analyzed by in situ hybridization and immunohistochemistry. Thirty-six animals received one subcutaneous injection of gentamicin sulfate (50 mg/kg) daily for ten days. The animals were allowed to survive 1, 3, 7, 14, 21 or 28 days (n = 6 in each group) before sacrifice and preparation for both in situ hybridization and immunohistochemistry. Twelve animals were sacrificed as control for each groups (n = 2). RESULT: The results showed that the supporting cells and cells in the lumenal nuclear layer within the damaged region were strongly labeled for c-myc transcripts and its protein. Seven days after last injection, the highest level of c-myc mRNA and c-myc protein expression was observed in the injured sensory epithelium. C-myc gene expression began to decline from fourteen days post injection. High level expression of c-myc gene was discussed in relation to the time and location of HC regeneration. CONCLUSION: The results indicate that c-myc gene expression may be an important molecular event in HC regeneration and may play a role in regulating progenitor cells proliferation. PMID- 12563998 TI - [Use PCR technique to detect mitochondrial DNA in rat membranous labyrinth]. AB - OBJECTIVE: To establish a sensitive and reliable method of mtDNA detection in the membranous labyrinth of rat inner ear. METHOD: A mtDNA segment of 601 bp containing the genes which encode ND1 subunit 16S rRNA in rat inner ear membranous labyrinth was detected by PCR method. The method is compared with Edris's mtDNA isolation method. RESULT: A 601 bp PCR product of mtDNA in the membranous labyrinth of rat ear inner can be obtained by the two methods. But the requirement of the sample quantities was different for the two methods. The extraction of mtDNA need two of inner ears membranous labyrinth to get reliably result by the method adapted from Seidman. However, the mtDNA extraction procedure from Edris's method need six of inner ears of the in order to get reliably result. CONCLUSION: The method of mtDNA isolation, amplification and detection membranous labyrinth of rat inner ear by PCR method adapted from seidman is more sersitive. PMID- 12563999 TI - [Aminoglycoside ototoxicity associated with mitochondrial DNA mutation]. AB - OBJECTIVE: To confirm the effect of genetic susceptibility to aminoglycoside ototoxicity. METHOD: Mitochondrial DNA (mtDNA) from 62 members of 9 aminoglycoside induced deafness families was analysed by PCR-Restriction endonuclease digestion. RESULT: The nucleotide 1555A-->G mutation in 12S rRNA gene of mtDNA was identified in 20 members of 5 families. CONCLUSION: These findings indicate that this mutation is a molecular basis for genetic susceptibility to the ototoxic effect of aminoglycosides. The relationship between the audiovestibular changes and genetic susceptibility to aminoglycoside ototoxicity and the pathogenetic mechanism of susceptible deafness are discussed. The possibility that other gene mutations can also predispose to aminoglycoside ototoxicity is proposed. PMID- 12564000 TI - [Combined window genyantrum in middle and inferior meatus under sinus endoscopy]. AB - OBJECTIVE: For clearing away process of genyantrum and preventing its recurence. METHOD: 36 patients (40 sinuses) were treated with combined window in middle and inferior nasal meatus under sinus endoscopy. The process was cleared away completely. RESULT: There is no complication after one year following up. The cure rate is 100% and the recurrence rate is 0. CONCLUSION: It shows that the method has the advantage of clearing away the process of genyantrum and preventing recurence. PMID- 12564001 TI - [Treatment for chronic dacryocystitis by probing through naso-lacrimal duct under endoscopy]. AB - OBJECTIVE: To determine the possibility of curing chronic dacryocystitis by probing through naso-lacrimal duct under endoscopy. METHOD: 30 adult eyes with chronic dacryocystitis were divided into two groups: 20 treated under endoscopy as endoscopy group and 10 under rhinoscope as control. RESULT: In endoscopy group, treatment in all 20 eyes successed at the first time, but in control group only in 8 of the 10 eyes successed at the first time and the other 2 needed second surgery. CONCLUSION: Treatment for chronic dacryocystitis by probing through naso-lacrimal duct under endoscopy is better. PMID- 12564002 TI - [Diagnosis and prognosis evaluation of primary non-Hodgkin's lymphoma of the nose]. AB - OBJECTIVE: To Study the diagnosis of primary nasal non-Hodgkin's Lymphoma (NHL) and to find the relationship between its classification, stage and prognosis. METHOD: The data of 10 cases with primary nasal NHL documented pathologically were analysed retrospectively. RESULT: The classification and stage were related to the prognosis significantly. Following up seven cases, 5T-ML with severe systemic symptoms died 4-6 months after diagnosis and treatment. One earlier T-ML is survival till now, and another B-ML was alive for 2.5 years. CONCLUSION: Early diagnosis of NHL of nose based on pathology is very important for treatment and prognosis. Repeat biopsies will be necessary when the diagnosis is suspected. The prognosis of late T-ML is disastrous. PMID- 12564003 TI - [Clinical use of nasal airflow measurement in septoplasty]. AB - OBJECTIVE: To discuss the clinical use of nasal airflow measurement in septoplasty. METHOD: After nasal decongestion, 21 patients's nasal airway resistance and nasal airflow were evaluated by anterior rhinomanometry before and after submucous resection of nasal septum. RESULT: The amount of nasal airflow asymmetry could show nasal respiratory function better than nasal airway resistance does, the patients with absolute amount of nasal airflow asymmetry greater than 50% had bad postoperative outcome, patients' satisfication with operation was significantly correlated with the presence of post-operative symmetrical nasal airflow. CONCLUSION: The amount of nasal airflow asymmetry can serve as an indicator of patient selection, outcome evaluation and prognosis in nasal septum deviation. PMID- 12564004 TI - [Analysis of T cell subgroup in children with allergic rhinitis]. AB - OBJECTIVE: To observe the different distribution of T cell in peripheral blood between normal children and children with allergic rhinitis. METHOD: We use the indirect immunofluorescence to make a statistical analysis randomly in 16 children with allergic rhinitis and 20 in control group. RESULT: CD3, CD4, CD8 in peripheral blood of patients are a lot less than those in normal children. CONCLUSION: The change of T cell subgroup's distribution is an important process of immune mechanism disturbance and can be used as a monitoring standard in allergic diseases. PMID- 12564005 TI - [Isolated sphenoid sinus disease]. AB - OBJECTIVE: The purpose of this study is to review isolated sphenoid sinus disease, to establish correct diagnosis and to treat it effectively in the future. METHOD: 26 patients underwent endoscopic surgery, in which 20 sphenoid sinus mucocoeles were removed and 6 sphenoid sinuses were explored. 3 sphenoid sinus mucocoeles were removed by external approach, 1 patient underwent skull base surgery and decompression of optic nerve. RESULT: 24 patients were free from disease postoperatively within 0.5-4 years follow-up, 2 patients underwent re operation because of recurrence, 1 patient was undergoing radiotherapy, 3 patients could not contact. CONCLUSION: This study shows that CT not only makes contribution to establish correct diagnosis but also plays an important role in surgery of isolated sphenoid sinus disease. Endoscopic surgery is valuable in treatment for benign disease of sphenoid sinus. PMID- 12564006 TI - [Endoscopic pituitary tumor surgery by nasal cavity and sphenoid sinus]. AB - OBJECTIVE: For reducing surgical trauma to patients of pituitary tumor. METHOD: 12 patients of pituitary tumor were treated with endoscopy by nasal cavity and sphenoid sinus under general anaesthesia. RESULT: Except one died of bleeding within cranial cavity after operation, the rest recovered smoothly without severe complications. During 7 months to one and half year follwing-up, the function of endocrine in 9 cases resumed to normal and 8 of their vision improved to some extent. CONCLUSION: The good illumination and clear visual field under endoscopy could improve the accuracy and security of operation and minimize the injury to normal tissue. But for large or recurrent tumors, this approach should be selected carfully. PMID- 12564007 TI - [Effect of postoperative treatment on endoscopic sinus surgery]. AB - OBJECTIVE: To investigate the effect of postoperative treatment on endoscopic sinus surgery (ESS) and to discover the best way of care. METHOD: We checked 54 patients who had undergone ESS and been followed-up. RESULT: 7 cases (13.0%) were found with recurrent nasal polyps; 8 cases (14.8%) with recurrent sinusitis; 7 cases (13.0%) with adhesion and 10 cases (18.5%) with incomplete epithelialization. CONCLUSION: A suitable postoperative treatment and reasonable regular following-up by endoscopy is one of the important segment for securing optimal long-term result. PMID- 12564008 TI - [Study on relative factors in endoscopic sinus surgical procedure]. AB - OBJECTIVE: In order to investigate the factors which affect the operative treatment effect of FESS. METHOD: 215 cases suffered functional endoscopic sinus surgery (FESS) from May 1994 to December 1997 were analysed. RESULT: The clinical curative rate was 75.8% and the improving rate was 96.3%. CONCLUSION: The effect shows that surgical procedure is closely related to following factors: mechanic of operation, drugs taken in pre-operation, kinds of anesthsia, clinical stage of chronic sinusitis, nasal poplys and skill of surgeons. PMID- 12564009 TI - [Variations of the middle meatus mucosa ultrastructure after endoscopic sinus surgery]. AB - OBJECTIVE: To explore the variations of middle meatus ultrastructure after endoscopic sinus surgery. METHOD: 10 cases whose middle meatus mucosa recovered well after undergoing endoscopic sinus surgery were screened, and postoperative electron microscope examinations of middle meatus mucosa were made in the first, second, third and forth month. RESULT: In the first month after operation, there was obvious fibre proliferation in the mucosa. In the second month, there was fibre proliferation and dissolution existing simultaneously. In the third month, the mucosa's structure resumed fundamentally to normal, but its function was not perfect. In the forth month, the mucosa was completely normal and can function normally. CONCLUSION: The mucosa of middle meatus can resume completely after endoscopic sinus surgery but need at least four months. It is very important to have a long time reexamination for improving therapeutic effect. PMID- 12564011 TI - [Glottal configuration and histological findings in vocal muscles following trauma to cricoarytenoid joint]. AB - OBJECTIVE: The unilateral cricoarytenoid joints of the rabbits were traumatized bluntly to investigate the changes of glottal configuration and histology of the vocal muscles. METHOD: 10 New Zealand white rabbits divided into 4 groups were studied. Under direct laryngoscope, the unilateral cricoarytenoid joint of each rabbit was traumatized bluntly. The glottal configuration and histology of the true cords demonstrated atrophy of the vocal muscles. The remarkable atrophy of the vocal muscles was evaluated after 1, 3, 6 and 9 months. RESULT: In 1 and 3 months after trauma, the glottal configuration of the rabbits showed no difference from that in the period of trauma. The histology of the vocal muscles was approximately normal. The lateral drift and deformity of the fixed vocal cord and wider glottal gap were seen 6 months after trauma. The histological sections of the vocal muscles were noted 9 months after trauma. CONCLUSION: The atrophy of the vocal muscles may occur 6 months after trauma to cricoarytenoid jonint. This pathologic change may cause the lateral displacement and deformity of the vocal cord, which increases the glottal incompetence and affects voice recovery. The laryngeal electromyography may be influenced by the pathologic changes of the vocal muscles. PMID- 12564010 TI - [Clinical and bacteriology observation on intraoperation usage of tarivid otic solution in treatment of chronic otitis media with cholesteatoma]. AB - OBJECTIVE: To determine the effect and safety of Tarivid (oflorxacin) otic solution used during the operation in treatment for chronic otitis media with cholesteatoma. METHOD: 42 patients were divided into two groups. In Tarivid group, 23 patients' operative cavities were soaked with 0.3% Tarivid, while other 19 patients' operative cavities in boric acid group were soaked with 4% boric acid during the open mastoidectomy with tympanoplasty. RESULT: The recovery rate was 82.6% and 50% respectively. There was significant difference between two groups (P < 0.05). No side effect was observed. CONCLUSION: It shows that using Tarivid otic solution in the operation can improve the outcome of the operation and has high security. PMID- 12564012 TI - [The effect of granule-rhinitis on allergic nasal mucosa]. AB - OBJECTIVE: To investigate the effect of granule-rhinitis on allergic rhinitis. METHOD: Experimental guinea pigs with allergic rhinitis were treated with granule rhinitis. During the total course of treatment, sneezing and nasal itching were observed and recorded and then compared quantitatively with those of the control groups. Histological changes of the nasal mucosa were observed. RESULT: The various symptoms of allergic rhinitis were obviously relieved in the treatment group. Granule-rhinitis reduced inflammatory cell infiltration, particularly decreased the number of mast cells and eosinophils in the superficial layers of the nasal mucosa, and reduced the release of mediators from mast cells. It diminished the vasodilatation in nasal mucosa. CONCLUSION: Granule-rhinitis is effective in the treatment of allergic rhinitis. PMID- 12564013 TI - [Study on the nasal resonance]. AB - OBJECTIVE: In order to understand the resonant function of the nasal cavity (including nasal sinuses) which is one of the resonant organs of the human body. METHOD: The maximum frequency, mean frequency and the number of harmonics of five vowels [a], [e], [i], [o], [u] were compared before and after packing of the anterior nasal cavity. RESULT: The result was analyzed statistically and showed no significant difference. CONCLUSION: It proved that the front part of the nasal cavity plays no important role in the function of the nasal resonance. PMID- 12564014 TI - [CT scanning analysis for maxillary sinus bony septum abnormalities and its clinical significance]. AB - OBJECTIVE: To assess the extent and distribution of disease as well as associated anatomic abnormalities in patients with chronic sinusitis and nasal polyps. METHOD: Both axial and coronal CT scans of eight patients who failed medical therapy and functional endoscopic sinus surgery (FESS) were reviewed retrospectively. RESULT: Among them, five were found to have vertically aligned coronal bony septum, two were horizontal, one sagittal. Complete bony septum divided the maxillary sinus into two separate cavities in four patients, others had incomplete bony septum. Based upon the CT scan findings, patients suffered from these anatomic abnormalities received Galdwell-Luc procedures and pansinusectomy through mid-facial degloving approach respectively. All patients did well postoperatively and were symptom free for six months. CONCLUSION: The results showed that the CT scan can display the distribution and extent of residual disease, demonstrate anatomic abnormalities, and guide therapeutic intervention. The combination of axial with coronal CT scan can effectively determine the incidence of the maxillary sinus bony septum. PMID- 12564015 TI - [Microwave therapy of radiotherapeutical choanal atresia under nasal endoscope]. AB - OBJECTIVE: In order to know efficacy of microwave to treat the secondary choanal atresia under nasal endoscope. METHOD: Three patients suffering from membranous choanal atresia due to radiotherapy of nasopharyngeal carcinoma (NPC) were treated by microwave under nasal endoscope. RESULT: Satisfing recoveries were accomplished in all patients and no relapse was found within the 7-13 months' follow-up. CONCLUSION: This method has many advantages, such as: processing available under local anesthesia without incision, mild damage to normal tissues, clear operation visual field, less hemorrhage and postoperative pain, complete coagulation and gasification of atresic membrane, no need of postoperative dilation and stable therapeutical effect. PMID- 12564016 TI - [Prevention of complications of endoscopic sinus surgery]. AB - OBJECTIVE: To investigate the causes and prevention of the complications of endoscopic sinus surgery (ESS). METHOD: We summurized the complications of 26 cases among 100 patients who were performed the ESS operations. RESULT: The complications included hemorrhage, cerebrospinal fluid leak, orbital hematoma, subcutaneous orbital emphysema, synechia and antrotomy closure. CONCLUSION: We concluded the factors which probably result in complications and the prevention of complications. PMID- 12564017 TI - [The experience of treatment of 7 cases of olfactory neuroblastoma]. AB - OBJECTIVE: To improve the treatment result of olfactory neuroblastoma. METHOD: Introduce the experience of treatment of 7 patients with this disease. Among these 7 cases, 2 of them were stage B while the others were stage C according to the system proposed by Kadish. 3 cases received radiotherapy alone, and 4 others received radiotherapy pre-operation. The radiation dose was 5000-7000 cGy. All the patients were followed-up until December 1997. RESULT: 5 cases are still alive, survival time are 20, 22, 23, 29 and 76 months respectively. The other 2 patients died with the survival time of 6 and 48 months respectively. CONCLUSION: Radiotherapy seems effective in the treatment of olfactory neuroblastoma; the combination of chemo-radio-therapy is preferable for advanced or metastasis olfactory neuroblastoma. We extrapolate there are different subtypes on pathology in this tumor. PMID- 12564018 TI - [The applied anatomy and measure of nasolacrimal duct]. AB - OBJECTIVE: To know the anatomy of nasolacrimal duct. METHOD: 80 half-skulls of chinese adult were used to observe shapes of the orifice. RESULT: Five shapes of the orifice were found. Slit-shape was the most in all shapes. 73 nasolacrimal ducts and situation of orifices have been measured with results as follows: (1) The nasolacrimal duct length is 14.14 mm (mean value); (2) The distance from the inferior margin of the anterior nostril to the anterior margin of the orifice (D.N.O.) is 29.00 mm (mean value); (3) The distance from the anterior attachment of the inferior turbinate to the anterior margin of the orifice is 11.70 mm (mean value); (4) The distance from the inferior margin of the anterior nostril to the anterior attachment of the inferior turbinate (D.N.T) is 18.30 mm (mean value). With analysis of linear regression, the relationship between the "D.N.T." and the nasolacrimal duct length, the "D.N.T." and the "D.N.O." is positive linear correlation respectively, "D.N.T." has been able to be measured easily, so the nasolacrimal duct length and the situation of the orifice have been recognized. CONCLUSION: D.N.T. may be used in clinical teaching and researching work. PMID- 12564019 TI - [Study of rhinophysiology for normal young adult in the Cantonese of China by acoustic rhinometer]. AB - OBJECTIVE: In order to study the site of nasal parameters, nasal minimal cross sectional area (NMCA), nasal cavity volume (NCV), nasal resistance (NR) and the relations among them. METHOD: 276 younger adult of normal Cantonese were tested by ECCOVISION acoustic rhinometer. RESULT: The results were as follows: 72.95% NMCA in subjects are positioned at internal nostril. The distance from the position of NMCA to the nostril is 1.36 +/- 0.53 cm in male, and 1.06 +/- 0.46 cm in female. There was no difference in NMCA, NCV and NR between male and female. There was significant relationship between the NR and NMCA. There was relationship between the NR and NCV, and there was little relationship between the NR and DCAN. CONCLUSION: Acoustic rhinometry is a new objective rhinometric method, but different race must have its standard respectively. PMID- 12564020 TI - [Knowing from experience of surgical treatment to 27 cases of facial nerve injures]. AB - OBJECTIVE: In order to investigate the elements which affect operative treatment effect to patients with facial nerve injuries. METHOD: 27 postoperative cases with facial nerve injures were analyzed. RESULT: The results showed that the effect of treatments was more satisfactory when age was younger, the timing of operation was earlier, operative approach was right, facial nerve injury was mild, or lesion position was lower. The effect of postoperation reached House I II degree in 21 patients (21/27), House III-IV degree in 6 patients. CONCLUSION: According to different causes of facial nerve injuries, the keys to efficient treatment are confirming of injury range, selecting of operative approach, probing into damages completely in case of missing lesion. PMID- 12564021 TI - [Diagnosis and restitution of nasal bone fracture]. AB - OBJECTIVE: To discuss accurate methods of diagnosis and restitution for nasal bone fracture. METHOD: 280 cases of nasal trauma were examined by lateral x-ray test (horizontal CT-scan was also used in 76 cases of these patients). RESULT: 220 cases were found with bilateral fracture of nasal bone. Unilateral fracture, simple fracture of frontal process of maxilla, compound fracture of nasal bone and frontal process of maxilla, compound fracture of nasal bone, maxilla and perpendicular plate of ethmoid bone were seen in 12, 15, 30 and 3 cases respectively. CONCLUSION: X-ray examination can only find bilateral fracture of nasal bone, but CT-scan can demonstrate unilateral fracture, fracture of frontal process of maxilla and fracture of perpendicular plate of ethmoid bone. Restitution with pincers is still a routine therapeutic method for nasal bone fracture at present, and nasal septal reconstruction is necessary when both nasal bone and perpendicular plate of ethmoid bone were involved. But for those patients with high-position compound fracture, restitution by directly visual operation through lip-dental-gum approach is the best choice. PMID- 12564022 TI - [Operative approach choosing to treat cholesteatoma otitis media (with clinical analysis of 860 cases)]. AB - OBJECTIVE: In order to improve operative effect to cholesteatoma otitis media. METHOD: The clinical data of 860 cases of patients with cholesteatoma otitis media were treated by surgery through three approachs (through epitympanum, cribriform area or combined approach) were analyzed. RESULT: The results showed that the recurrent rates of them were 10%, 11% and 6% respectively; The rates of improvement of hearing were 56%, 40% and 69% five years after operation respectively, and after 5 to 15 years were 45%, 35% and 62%; The rates of different abnormal shapes of tympanic membrane (including retraction, adherence and perforation) were 43%, 47% and 12%. CONCLUSION: According to the location of foci, we regard that various and appropriate operative approach should be selected to treat the cases with cholesteatoma otitis media, and the combined approach tympanoplasty has advantage over the other two. PMID- 12564023 TI - [Experimental and clinic study of the maxillary sinus plasty following Caldwell Luc]. AB - OBJECTIVE: To investigate the clinical effect of surgical procedures on patients with chronic maxillary sinus disease. METHOD: On the basis of experimental study with animal, this prospective study included 63 patients with chronic maxillary sinus disease who had a maxillary sinus plasty following Caldwell-Luc. The operation procedures included anterior wall ring-drilling, and middle meatal antrostomy as well as sinus wall plasty. RESULT: Experimental results showed that the repair and regeneration of maxillary sinus mucosa was intact in dogs after 5 weeks of radical operation. Clinical study indicated that the condition of anterior wall with a resutured bone-lid was normalized radiographically during the follow up time of 1-3 months. The patency rate of the middle meateal antrostomy 18-42 months post-operatively (average 28 months) is 96%. CONCLUSION: We conclude from the study that the maxillary sinus plasty recovers anatomic construction, maintains physiological drainage way and provides a good environment for repair and regeneration of the sinus mucosa. PMID- 12564024 TI - [The utilization of bronchoalveolar lavage in children]. AB - OBJECTIVE: To investigate the clinical effect of bronchoalveolar lavage (BAL) in children with atelectasis owing to pulmonary infection. METHOD: We compared the preactive period in test group (87 cases) who were treated by BAL with control group (30 cases) who were treated by conventional drug. RESULT: The result showed a shorter preactive period in test group than that of control group (P < 0.01). No significant difference of effect of treatment was found between this two groups (P > 0.05). CONCLUSION: Under directly visual metal bronchoscope, we can improve the ventilation function by eliminating the obstructions, and can fasten the absorption and recovery of the foci by using antibiotics locally. So BAL can obviously shorten the course of treatment. Due to the structure defect, using of BAL with fiber bronchoscope is restricted in children. However, metal bronchoscope under total anesthesia can provide a good operation condition which include a large ventilation cavity and a wider visual field, so it has high clinical value in children. PMID- 12564025 TI - [Diagnosis and treatment of Killian's polyp under nasal endoscope]. AB - OBJECTIVE: To further understand the advantage of examination and operation for killian's polyp under nasal endoscope. METHOD: 22 cases with Killian's polyps were diagnosed and treated under nasal endoscope. RESULT AND CONCLUSION: Before operation, examination under nasal endoscope may clearly find origin of the polyp and foci in maxillary sinus, which plays more important role in differential diagnosis and operation than CT or X-ray test. Operative treatment of Killian's polyp under nasal endoscope can not only eradicate foci but also reserve function of mucosa, but for severe foci in the maxillary sinus, the optimum operation should be Caldwell-Luc antrotomy. PMID- 12564026 TI - [Use of diagnostic ultrasound of sinus in the paranasal sinus disease]. AB - OBJECTIVE: To utilize the diagnostic A ultrasound of sinuses in the diagnosis of paranasal sinus disease. METHOD: Through history enquirement, careful clinical otorhinolaryngological examination, X-ray examination of paranasal sinus, CT, maxillary sinus puncture and examination of the diagnostic ultrasound of sinus, we observed 115 patients with paranasal sinus disease and compared them with 22 normal people who were in control group. RESULTS: In normal people, the according rate between ultrasound examination and clinicaly diagnostic methods, include history enquirement, otorhinolaryngological examination, X-ray examination, CT and maxillary puncture, is 100%. And in the group of patients with paranasal disease, the according rate is 62.5%-92%. CONCLUSION: The diagnostic A ultrasound of sinus has high accuracy, the control is easy, the cost is relatively lower, it can be used repeatedly and do no harm to the patient's health. It is a good method and assistant technigue in the diagnosis of frontal and maxillary sinus disease. PMID- 12564027 TI - [Experimental study on carotid artery resection with vascular replacement by an autogenous vein graft]. AB - OBJECTIVE: In order to observe the survival rate and the histologic changes of the reestablished common carotid artery system. METHOD: A planned resection of the carotid artery was done and the common carotid artery system was reestablished by interposing an autogenous external jugular vein graft in 15 white rabbits. RESULT: 12 survived, the other 3 rabbits died from graft thrombosis, the survival rate was 80%. Histologic examination demonstrated the smooth internal tunic in the anastomotic part of the carotid-jugular and no graft thrombosis. Angiography demonstrated the free circulation of the blood and no aneurysm in the anastomotic vein. CONCLUSION: It is concluded that it is possible that in selected patients, a planned resection of the carotid artery with replacement by interposing an autogenous external jugular vein graft will be done. PMID- 12564028 TI - [Clinical value of distortion product otoacoustic emission (DPOAE) and their contralateral suppression effects]. AB - OBJECTIVE: The present paper aimed at examining the role of DPOAE and their contralateral suppressive effects in evaluation medial olivocochlear system (MOCS) function status in human. METHOD: Seventeen subjects (34 ears) with normal hearing, 13 ears with cochlear hearing loss and 9 ears with retrocochlear hearing loss were studied on their DPOAE and contralateral suppression. RESULT: The results revealed that DPOAE amplitude significantly reduced in ears with cochlear hearing loss, contralateral suppression was smaller than normal ears but there was no significant difference between two groups. By contrast, DPOAE amplitude was much higher in ears with retrocochlear hearing loss than that in the other two groups. Contralateral suppression was significantly reduced or disappeared, and contralateral acoustic stimulus (CAS) even increased DPOAE amplitude in ears with retrocochlear hearing loss. CONCLUSION: DPOAE and suppression induced by CAS can be used as an index of medial olivocochlear system (MOCS) function in patients with sensori-neuaral hearing loss, and can differentiate cochlear hearing loss from retrocochlear hearing loss. The tests of DPOAE and contralateral suppression may have much more advantages in clinical application over any other audiological test. PMID- 12564029 TI - [Middle ear operation of children and juveniles]. AB - OBJECTIVE: To analyse the results and effective factors of middle ear operation in children and juveniles. METHOD: 105 cases (146 ears) were analyzed retrospectively. 49 ears were followed up over 3 months. RESULT: 120 ear had perfect hearing materials. The difference of average air-bone gap between postoperation (21.86 +/- 13.15) dB HL and preoperation (35.20 +/- 13.87) dB HL was significant (P < 0.01). CONCLUSION: Middle ear operation was a safe, effective operation in children and juveniles, reoperation also was proper option for someone if his first operation didn't success or repeated cases. PMID- 12564030 TI - [Transformation of effusion and formation of granulation tissue in the pathologic process of otitis media with effusion]. AB - OBJECTIVE: To explore the transformation of stagnant effusion (SE) and the pathologic behavior leading to granulation tissue (GT) formation in the pathologic process of otitis media with effusion (OME). METHOD: The temporoal bone slides from 306 ears with OME at the Otopathology Laboratory of the University of Minnesota USA were studied histopathologically under light microscopy. RESULT: The observations revealed a pathologic process in which the type and the condition of stagnant effusion in the middle ear cleft was variable in the dynamics of OME progression from an early pathologic stage to an advanced pathologic stage. The granulation tissue was primarily formed by two types of fibroblast proliferation in subepithelial space. The absorption and the orgnization of SE and the formation of granulation tissue maybe occur simultaneously in same areas and a same pathologic process. CONCLUSION: The granulation tissue can only form in the areas where effusion has stagnated and has been absorpted. It is very important that effusion of the otitis media should be removed completly before the formation of granulation tissue. PMID- 12564031 TI - [Adhesive otitis media with cholesteatoma]. AB - OBJECTIVE: To explore the mechanism of cholesteatoma formation in adhesive otitis media with cholesteartoma. METHOD: Three cases of this kind of disease were reported and literatures were also reviewed. RESULT AND CONCLUSION: 1. It is a acquired primary cholesteatoma. 2. The eustachian tube and tympanic isthmus block are the direct cause of these cholesteatoma. 3. CT examination of the temporal bone is more valuable in diagnosis than that of normal X-ray. 4. If the cholesteatoma lesions can be removed completly, tympanoplasty may be performed in first stage or separate stage. PMID- 12564032 TI - [Ultraviolet blood irradiation therapy for the sudden deafness]. AB - OBJECTIVE: In order to improve curative effect of sudden deafness. METHOD: We adopted ultraviolet blood irradiation (UBI) therapy added conventional pharmacotherapy to treat 70 cases of sudden deafness. RESULT: The general effective rate was 85.6% and there was no complication. In control group we adopted conventional pharmacotherapy only, the general effective rate was 65.7%. There was significant difference between experimental and control groups (P < 0.01). CONCLUSION: The results showed that the UBI is a safety, effective, simple and convenient therapy for the sudden deafness. PMID- 12564033 TI - [The use of autogeneic residual auditory ossicles, allogeneic auditory ossicles and the allogeneic nasoseptal cartilage in tympanoplasty]. AB - OBJECTIVE: In search of ideal substance(s) for middle ear reconstruction, various materials were used and investigated in tympanoplasty. METHOD: Up to may 1996, three kinds of materials were be used to reconstruct the ossicular chain: 1. Auto ossicular residue reposition, 153 ears; 2. Homo-ossicle transplantation, 119 ears; 3. Homo-chondroseptum transplantation, 93 ears. We analysed and compared the middle and long-term results in these three groups. RESULT: 1. In auto ossicular residue reposition group, 86.3% was satisfied 10 years postoperation; 2. The satisfaction rate was 76.5% in homo-ossicle transplantation group; and 3. In the group of homo-chondroseptum graft, 77.4% of satisfaction rate was reached. CONCLUSION: Auto-ossicular residue reposition is a better choice for ossicular chain reconstruction. That is responsible for its convenient, free of rejection, and better long-term results. PMID- 12564034 TI - [Diagnostic value of ENG in occupied lesions of cerbellopontine angle]. AB - OBJECTIVE: To evaluate the diagnostic value of ENG in occupied lesions of cerebellopontine angle. METHOD: Twenty-four cases of pathological confirmed occupied lesions of cerebellopontine angle were examined with ENG before operation. The ENG examination included saccade test, gaze test, ocular pursuit test, optoknetic test, positional test and positioning test. RESULT: ENG tests were compared with the results of operation and pathology. The results showed that abnomal ENG-OKR suggested central lesions. Brun's nystagmus and unilateral canal paresis were of important value in locating the lesions of cerebellopontine angle. The ocular pursuit test and optokinetic test could point out which side the lesions were. CONCLUSION: Patients with cerebellopontine angle lesions smaller than 3 cm in diameter showed unilateral canal paresis in caloric test indicated acoustic neuroma. Patients with cerebellopontine angle lesions showed no canal paresis may be other lesions except acoustic neuroma. PMID- 12564035 TI - [Complications of cerebellopontine angle surgery by retrosigmoid approach]. AB - OBJECTIVE: To know the management of complication of cerebellopontine angle surgery by retrosigmoid approach. METHOD: Fourteen cases of cerebellopontine angle surgery by retrosigmoid approach were analyzed retrospectively. RESULT: All patients have been cured except that one died. CONCLUSION: To know the causes and mechanism of the complications during operation and post-operation is very important for prevention and management of the complications. PMID- 12564036 TI - [Expression and clinical implication of multidrug resistant-associated protein gene (MRP) in patients with laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To study the relationship between the expression of MRP mRNA and the clinical pathological characteristics of the patients with laryngeal squamous cell carcinoma. METHOD: Reverse transcription polymerase chain reaction (RT-PCR) assay was used to detect the expression of MRP mRAN in 35 patients. RESULT: The positive expression rate of MRP mRAN was 45.7 percent (16/35) in laryngeal carcinoma. It was higher in advanced (T3-4) patients than that in early (T1-2) cases (P < 0.05) and showed a significant correlation with metastases of cervical lymph node (P < 0.01). CONCLUSION: It is suggested that the expression of MRP mRNA may not only play an important role in multidrug resistance of laryngeal carcinoma, but also influence their biological behavior of dissemination and metastases. PMID- 12564037 TI - [The submucosal structure of the endolymphatic sac of guinea pigs]. AB - OBJECTIVE: To investigating the submucosal structure of the endolymphatic sac (ES), so as to analyse the role of ES in the function of inner ear. METHOD: The temporal bone of the guinea pigs were cleared in methyl salicylate and inspected under a stereomicroscope. The ultrastructure of endolymphatic sac has been observed by transmission electron microscope. RESULT: The extensive vascular system around the sac and has compact contact with sinus sigmoid. Its submucosal space comprises both arterioles and venules, as well as lymphatic sinus. CONCLUSION: The result suggests that the ES is a very metabolically active structure and has a pressure regulating function. The disturbance of endolymphatic resorptive function seems to result endolymphatic hydrops after the vascular supply poverty of endolymphtic sac. It's may be the causative factor of Meniere's disease. PMID- 12564038 TI - [Hair cell apoptosis and hearing loss of perilymphatic fistula]. AB - OBJECTIVE: To investigate the effect of hair cell apoptosis on hearing loss of perilymphatic fistula (PLF) in guinea pig. METHOD: Twenty-five guinea pigs with light microscope and TdT mediated biotin dUTP nick-end labelling (TUNEL) techniques. The ECochG and ABR were measured and the data analyzed with statistics. RESULT: 1. The apoptosis of hair cell was not revealed in 0-hour group and 2-hour-group of PLF. Following the time of PLF was longer, the apoptosis of hair cell was more increased. The apoptosis of hair cell was demonsted in 1-day-group (1 case, 17%), 2-day-group (4 case, 67%) and 7-day-group (4 case, 80%); 2. The amplication of AP was reduced after operation induced PLF. The CAP of experimental ear was significantly higher than that of control ear (P < 0.01) in 1-day-group, 2-day-group and 7-day-group. CONCLUSION: 1. Apoptosis of hair cell was appeared in PLF; 2. The apoptosis of hair cell may be one of morphological evidence in hearing loss of PLF. PMID- 12564039 TI - [Gene therapy: new developments]. AB - Gene therapy is based on the transfer and the expression of therapeutic genes in specific target cells. For the treatment of genetic diseases gene therapeutic approaches aim at replacement of the deficient gene or at the correction of the genetic defect. Malignant diseases and an increasing number of other acquired diseases are additional targets for gene therapeutic strategies. For gene therapy to become a potential future treatment option, safe and therapeutically efficient gene transfer into specific target cells is a central requirement. A variety of nonviral and viral vector systems have been developed. Nonviral vectors transfer genes are far less efficient than viral vectors, but they have advantages due to their low immunogenicity and their large size capacity for therapeutic DNA. To improve the function of nonviral vectors, the addition of viral functions such as receptor mediated uptake, enhanced endosomal release and nuclear translocation of DNA may finally lead to the development of an artificial virus. In contrast, natural viruses are already highly developed structures for the transfer of nucleic acids. Recombinant viruses can be used for efficient gene transfer. Retroviruses, adeno-associated viruses and lentiviruses are suitable for gene therapeutic approaches which are based on the permanent expression of therapeutic genes such as the correction of enzyme deficiencies or the manipulation of hematopoetic stem cells, as they are able to integrate nucleic acids into the cellular genome. In contrast, adenoviral vectors result in highly efficient, but transient gene expression and are therefore especially useful for the treatment of malignant tumors. Novel developments of viral vectors mainly aim at the reduction of immunogenicity, increase of capacity for therapeutic genes and at improved vector production. Viruses which replicate selectively in tumor cells leading to tumor cell lysis represent a novel generation of viral vectors, which can further be improved by the addition of therapeutic genes resulting in enhanced tumor toxicity. PMID- 12564040 TI - [Immunotherapy: new insights]. AB - Immunotherapy of cancer, autoimmune diseases, and chronic viral infections is still in the early stages of development. This review deals with new immunotherapeutic approaches against tumors. Until now, a variety of specific and unspecific immunostimulatory strategies have been applied with only modest clinical success. The molecular characterization of novel human tumor antigens and the increased understanding of the immunological pathways involved in tumor immunity, however, have paved the way for the design of promising gene-based cellular cancer immune vaccines. Tumor antigens are readily available now and several ways to deliver antigen have been developed. The most important and widely used approaches consist of whole-cell vaccines, dendritic cell-based immunotherapy, and peptide vaccines. Clinical trials have, in general, demonstrated the safety of such strategies. There is, however, an important lack of information regarding to the correlation of clinical responses and analysis of the antitumoral immune effectors. Recently, exciting new immunological techniques and tools have been developed which allow to characterize antigen specific T cells at a single-cell level. In future, tumor rejection antigens have to be identified using new approaches such as reverse immunology which can be targeted therapeutically. A better quality control of the different immunotherapeutic approaches is mandatory if used in patients and the different therapeutic modalities need to be compared directly. This standardization of immunotherapeutic clinical trials may allow the identification of patient sub groups responding favourably to treatment. The precise definition of immune parameters will ultimately result in optimised therapeutic approaches. Finally, the identification of antigen specific T and B cells by new assessment tools may result in novel surrogate markers for clinical diagnostic immunotherapy. PMID- 12564041 TI - [Molecular virology of hepatitis C]. AB - Hepatitis C virus (HCV) infection is a leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma worldwide. Here, we will briefly review current concepts of the molecular virology of hepatitis C. In vitro and in vivo models of HCV replication will be discussed in this context. Finally, novel antiviral strategies will be outlined that result from an improved understanding of the viral life cycle. PMID- 12564042 TI - [Exacerbation of chronic hepatitis B after discontinuing of antiretroviral therapy in HIV infection]. PMID- 12564043 TI - [Benign recurrent intrahepatic cholestasis]. PMID- 12564044 TI - Evaluation of certain food additives and contaminants. AB - This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives and contaminants, with a view to recommending Acceptable Daily Intakes (ADIs) and tolerable intakes, respectively, and to prepare specifications for the identity and purity of food additives. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of food additives (including flavouring agents) and contaminants, assessments of intake, and the establishment and revision of specifications for food additives. A summary follows of the Committee's evaluations of toxicological and intake data on various specific food additives (diacetyltartaric and fatty acid esters of glycerol, quillaia extracts, invertase from Saccharomyces cerevisiae, beta-carotene from Blakeslea trispora, curcumin, phosphates, diphosphates and polyphosphates, hydrogenated poly-1 decene, natamycin, D-tagatose, carrageenan, processed Eucheuma seaweed, curdlan, acetylated oxidized starch, alpha-cyclodextrin and sodium sulfate), flavouring agents and contaminants (3-chloro-1,2-propanediol, 1,3-dichloro-2-propanol, and a large number of polychlorinated dibenzodioxins, polychlorinated dibenzofurans and coplanar polychlorinated biphenyls). Annexed to the report are tables summarizing the Committee's recommendations for ADIs of the food additives and tolerable intakes of the contaminants considered, changes in the status of specifications of these food additives and specific flavouring agents, and further information required or desired. PMID- 12564045 TI - Future trends in veterinary public health. AB - Since the publication of the report of a Joint FAO/WHO Expert Committee on Veterinary Public Health in 1975, many significant developments have occurred in this field. The present report of a WHO Study Group re-examines the role and functions of veterinary public health and its contribution to public health practice today and in the years to come. Since 1975, new, emerging and re emerging zoonotic diseases have acquired global significance for human health, and have required rapid responses from, and teamwork between, physicians, veterinarians, and biologists. The veterinary sector has a long distinguished history in contributing to the maintenance and promotion of public health. As health is multidimensional, health policy and practice should be interdisciplinary and intersectoral. Therefore, the contributions of other sectors--in particular agriculture, animal health and production, food industry, education, housing, public works, and communications--are vital. Such concerted action is particularly needed in developing countries with weak infrastructures and limited resources. This report reviews current and foreseen global changes for their potential implications on veterinary public health with regards to national and international policies, management of programmes and training. The report also provides recommendations for action in these areas. PMID- 12564047 TI - [Domestic, leisure activity and sports-related accidents in an active population: perspectives for prevention by health education]. AB - Resulting from the findings of epidemiological surveys on non-work-related accidents conducted in 1996 and 1997 among employees covered by a special health insurance programme for gas and electric company workers, this article identifies avoidable and recurrent accidents that may be targeted by health education interventions. It also emphasizes the importance of considering the needs expressed by the population at large when developing the themes for information campaigns. Finally, it proposes a sequel to the "survey-action plan" released in 1996. The proposed follow-up takes into account the occupational setting and its professionally-related barriers, and includes delivering prevention messages, conducting in-house experimental health promotion activities (and evaluating them), and disseminating intermediary external campaigns aimed at impeding avoidable accidents among adults. The preventable accidents identified in this study are not specific to the employed population group studied here; they may also be targeted by more general health promotion programmes and interventions. PMID- 12564048 TI - [Breast cancer prognosis in Tunisian women: analysis of a hospital series of 729 patients]. AB - The objective of this study was to identify the anatomical-clinical aspects and determine the prognostic factors for breast cancer in the central region of Tunisia. This retrospective study involved 729 patients suffering from breast cancer, proven either by histology or cytology, diagnosed and treated between January 1990 and June 1998 at the F. Hached University Hospital in Sousse, Tunisia. The patients' average age was 50 years (ranging from 22-91). The average size of the cancer at the time of diagnosis was 49.1 mm; 90% were invasive duct carcinoma with high histo-prognostic SBR grade (level II-III: 86%). The overall survival rate was 50.5% after five years, and 50% after seven years. Using univariate analysis, significant predictive value was found with the following factors: tumor size, the clinical ganglionic level, metastases at diagnosis, the number of nodes invaded, nodal capsular rupture and lymphatic embolism, SBR grade and the delay in seeking consultation. The multivariate analysis (Cox model) isolated two prognostic factors: the initial size of the tumor (p = 0.001) and metastases at the time of diagnosis (p = 0.01). The study's results indicated that breast cancer prognosis in Tunisia remains poor primarily due to late diagnosis. PMID- 12564049 TI - [Accreditation knowledge acquired by personnel in the Marseille hospital]. AB - Sensitization and educational training programmes are important pre-requisites in order to ensure the understanding of the issues at stake in the accreditation process. According to the preparatory research involved for this work, there has been no study published nor is there any documentation available on such a topic, specifically on carrying out the sensitization process to successful completion. The aims of this study are to evaluate the effectiveness of the sensitization phase and to refine general recommendations to serve as a guide for health institutions and their communications policies. A transversal randomised study was conducted through the distribution of a questionnaire to 107 health professionals from 23 clinical services in the public hospital system in Marseille. The results demonstrate that the knowledge about accreditation seems to be well integrated, with only 7 of the professionals being unaware of the accreditation programme, and 58% of them associating the accreditation process with an administrative procedure. Grouping the staff's responses according to professional category has shown to have almost no influence on the results. The level of overall knowledge is greater in more highly trained personnel (p < 0.05), but there was a poor level of knowledge regarding the internal organisational structures that existed. 75% of the health professionals thought that communication about accreditation was insufficient. The study authenticated the positive benefits of communication and identified a certain number of stumbling blocks to avoid. Several recommendations are proposed. PMID- 12564050 TI - [Moroccan adolescence in an urban milieu]. AB - This article presents the methods used and compiled results of two studies conducted involving Moroccan adolescents from Casablanca. The first one was carried out with a representative sample of young people ranging from 13 to 19 years of age from the urban area of Casablanca. It established and presented the socio-demographic situational context, their relationship with the school, their working environment and their social environment. The second one focused specifically on high school students by attempting to assess their knowledge on health matters and their socio-cultural attitudes. This study highlights the risks associated with professional integration and acclamation. It confirms the importance of the family and the school in their relationship and educational roles. Finally, it underlines the need for structures adapted to the needs of young people, such as in the health sector, as well as adequate free time to listen to them, in general. PMID- 12564051 TI - [General practitioners' practices and attitudes in regard to suicidal behavior]. AB - As demonstrated at the recent Consensus Conference on the "suicidal crisis" held by the French Psychiatry Union and the National Agency for the Accreditation and the Evaluation of Care, suicide is recognised as a current public health problem. The general practitioner's role is situated on two levels: in a preliminary stage, the early identification of risk, and in a secondary stage, the provision of care at the time of a suicide attempt and then the provision of on-going follow-up care in partnership with other special services. The study aimed at evaluating general practitioners' practices when encountered with suicidal patients and assessing their management of suicidal behaviour, as well as difficulties met and their expectations. Responses to a questionnaire were received from 290 general practitioners in one regional department in France. The results revealed that two-thirds of the respondents considered the identification of suicidal tendencies as being difficult, in particular with regard to the detection of young people at risk. General hospitals are the most common structures where patients are referred to in the event of a suicide attempt, as a facility that is permanently open and capable of delivering ambulatory care. A majority of the general practitioners expressed the need for training which was focused on methods for the early identification of risk as well as the development of listening skills. PMID- 12564052 TI - [Prevention in adolescence and the biomedical model, past and present history]. PMID- 12564053 TI - [Educating through fear, an anti-tobacco campaign]. PMID- 12564054 TI - [An unbelievable but clear confession]. PMID- 12564055 TI - [Volunteers' opinions of their first humanitarian aide mission]. PMID- 12564056 TI - [Workplace education? A medical service to be tested in the field]. PMID- 12564057 TI - [Mental suffering: ... what can be learned from the patients?]. PMID- 12564058 TI - [Psychiatric nurses working in the social welfare program for a guaranteed minimum income: the meeting of health and social services]. PMID- 12564059 TI - [Female urology]. PMID- 12564060 TI - [Assessment of pelvic floor dysfunction. Terminology standardization]. AB - OBJECTIVES: To record the findings from pelvic floor examination in an understandable way that permits the indication of a proper treatment. METHODS: We review classic classifications, as well as genital prolapse stages and degrees. We also record the POPQ examination system which is the established standard nowadays. RESULTS: Based on different classifications we intend to set up a systematic examination starting at the anterior segment, following with central and posterior compartments without forgetting pelvic floor descent, anal sphincter and pelvic floor muscles evaluation. CONCLUSIONS: The evaluation of pelvic floor defects is complex, complicating the pathologic diagnosis in order to determine the proper treatment. The various attempts to standardise classifications find problems. Nevertheless, a detailed history, a systematic examination and multidisciplinary cooperation will be able to correct this dysfunction. PMID- 12564061 TI - [Obstruction of the lower urinary tract in women]. AB - OBJECTIVES: To review the literature on female bladder outlet obstruction. METHODS: Female lower urinary tract obstruction clinical and urodynamic diagnostic criteria are defined. RESULTS: The main etiologic processes are identified: 1) Obstruction secondary to pelvic organ prolapse; 2) Postsurgical obstruction; 3) Uncoordinated voiding; 4) Detrusor-striated sphincter dyssinergia; 5) Bladder neck primary obstruction. The diagnostic features and various available treatments depending on the etiology (pharmacological, reeducative or surgical) are commented. CONCLUSIONS: Bladder outlet obstruction in the female is an underdiagnosed process frequently, which presents with a clinical picture different than in males, and demands a careful study from the urologist in order to identify the anatomical or functional cause. It will always be important to keep a high level of suspicion, mainly after treatment failures for voiding syndromes or recurrent urinary tract infections. PMID- 12564062 TI - [Hyperactive bladder]. AB - OBJECTIVES: To perform an update on the issue of detrusor overactivity, its incidence and treatment. METHODS: We developed this article combining the most recent publications about current knowledge and concepts on detrusor overactivity, our own experience and the database from the Hospital Ramon y Cajal Urodynamics Unit. RESULTS/CONCLUSIONS: Detrusor overactivity is a high incidence entity resulting from different pathologies that alter detrusor muscle function and control mechanisms. Currently, there are various treatment options that permit us to treat this entity with very good results. PMID- 12564063 TI - [Urinary incontinence. Evaluation and classification methods]. AB - OBJECTIVES: To evaluate different methods for clinical evaluation of female urinary incontinence. METHODS: A through bibliographic review has been carried out with special attention to the diagnostic tests of greater importance currently. Additionally we present the various female incontinence classifications. RESULTS: There is a growing interest in the use of questionnaires for subjective evaluation of symptoms. Among the objective tests for evaluation and quantification of the urine leaks the frequency/volume charts and the diaper test are very useful. Video-urodynamics and the late contributions from ultrasonography must be highlighted in the field of imaging tests. From the Urodynamics point of view, abdominal leak point pressure plays an interesting role in the diagnosis of female's urinary incontinence. CONCLUSIONS: After proper clinical evaluation, reasonable use of diagnostic tests and urologists experience will allow to set an exact diagnosis to offer the right treatment to the patients. PMID- 12564064 TI - [Rehabilitation in female stress urinary incontinence]. AB - OBJECTIVES: To know the results of a pelvic floor muscle training program in the treatment of female stress urinary incontinence. METHODS: Information phase: patient is informed about the disease, her anatomy, the objectives to be reached and how to perform the exercises. Treatment phase: 6 weeks of visual and auditory biofeedback assisted pelvic floor exercises to develop the pelvic floor muscles. Follow-up phase: one, 3, 6 month and 1 year visits with control for muscle evolution, motivation reinforcement and improvement of the symptoms. RESULTS: 412 Women underwent pelvic floor rehabilitation in a 4 year period, 45.9% were cured, 38.8% improved and 15.3% were treatment failures. CONCLUSIONS: Pelvic floor rehabilitation can cure female urinary stress incontinence and diminishes the number of surgical procedures and hospital costs in the treatment of urinary incontinence. PMID- 12564065 TI - [Stress urinary incontinence. Integral repair of pelvic floor defects]. AB - OBJECTIVES: To analyse the importance of complete pelvic floor defects correction at the time of female stress urinary incontinence treatment. METHODS/RESULTS: We review the basic principles in the treatment of genital prolapse. It is required to treat the anterior, central and posterior compartments. We describe the various anterior vaginal prolapse correction techniques and their basics depending on the association or not of urinary incontinence. The techniques for central compartment and posterior wall (rectocele) correction are also analysed. The recurrent vaginal dome prolapse and closure procedures are mentioned. CONCLUSIONS: We confront a multidisciplinary pathology in which urologists, gynaecologists, proctologists and pelvic floor physiotherapists can coexist and work together. Due to the variety of problems to be solved, it is necessary to standardise both the terminology and treatment, either surgical or not. PMID- 12564066 TI - [Prolene mesh sling in the treatment of stress urinary incontinence. Integral treatment of pelvic floor anomalies. Long-term results]. AB - OBJECTIVES: To evaluate the results of a comprehensive treatment of female stress urinary incontinence combining prolene mesh sling and proper gynaecologic repair depending on the kind of prolapse or pelvic floor dysfunction. To analyse short and long term clinical and urodynamic outcomes, and the effect on quality of life and economics associated with female urinary incontinence. METHODS: Prospective study including 102 consecutive patients with urinary incontinence; recruitment started in June 1996, ended in March 2002 for this analysis but it continues open currently. History of neurourologic disorders, radiotherapy, oncological diseases, gynaecological diseases and previous surgeries data were recorded in all subjects. History and physical examination were done evaluating urinary symptoms, duration of urinary incontinence, and urinary symptoms oriented examination (incontinence, urgency and urgency-incontinence), as well as gynaecological examination evaluating and grading cystocele, rectocele, uterine prolapse, enterocele and dome prolapse. Complete urodynamics were performed before and after surgery. Surgery was indicated as a complete pelvic floor dysfunction repair including prolene mesh sling in all cases with urinary stress incontinence, hysterectomy or not depending on the existence of prolapse, and anterior/posterior colpoperineorrhaphy with or without mesh. Results on urinary continence, complications and their treatment were evaluated in the postoperative period, on discharge, at 6 months and yearly thereafter. RESULTS: Average age was 63.8 years (27-82 years, SD 11.2). 39.3% of the patients were over age 70. Mean follow-up was 4.25 years (12-75 months, SD 11.9). The cost of pads for urinary incontinence was 2741.17 Euros per patient (456,117 pesetas). 32.3% of the patients had risk factors for urinary incontinence surgical treatment failure and 18.8% had a leak point pressure below 30 H20 cm. 22.3% cases presented with detrusor instability before surgery. 102 sling procedures, 20 hysterectomies, 26 anterior plasties, 14 posterior plasties, 10 mesh cystocele repair, 1 posterior mesh, and 2 enterocele sacral promontory fixation were performed, accounting a total of 173 surgical procedures during 102 anaesthesia procedures. 9 additional procedures were necessary for the treatment of complications. Stress continence was achieved in 99.01% cases. In half of the patients with preoperative urgency incontinence it continued during the first postoperative year. 11 cases have postoperative bladder instability, 7 of which had it preoperatively and 4 were de novo. COMPLICATIONS: 5 cases needed sling section/reconfiguration because of excess tension (non effective sling). 3 cases needed sling tight stretching/reconfiguration because of less than adequate tension. 2 cases of accidental bladder performation were treated with primary closure and urinary diversion. One case of infection-abscess in the mesh left anchoring stitch was drained under local anaesthesia. There were 6 cases of suprapubic, inguinal and rectal pain (8.1%), in all of them it disappeared within 9 months. There were 2 cases of wound infection. CONCLUSIONS: The prolene mesh sling can offer long term cure for stress urinary incontinence in almost all cases (99.01%), including the complicated ones. 91.1% of the patients underwent one surgical procedure only, and 8.8% required additional procedures. Results stand the test of time with a clinical-urodynamic follow up of 4.25 years. The voiding urgency referred by 81% of the women with large prolapses is associated with demonstrated bladder instability in 63% of the cases. Voiding urgency as well as bladder instability disappeared in all cases but one, being this fact prolapse-correction dependent, so that pelvic prolapse correction plays a decisive role. De novo bladder instability is uncommon (3.9%) and appears randomly in this series. PMID- 12564067 TI - [Laparoscopic technique in the treatment of stress urinary incontinence]. AB - OBJECTIVES: To evaluate the usefulness of laparoscopy as a corrective technique for urinary stress incontinence in our experience. METHODS: We review 72 cases of urinary stress incontinence who were treated by laparoscopic bladder neck suspension between April 1996 and December 2001. RESULTS: Average operative time was less than 40 minutes and mean hospital stay was 1.6 days. The continence rate was 82% after a mean follow up of 32 months. From the 12 patients with incontinence recurrence, 7 had the urinary incontinence recurrence within 2 months from surgery. CONCLUSIONS: The results of laparoscopic correction in urinary stress incontinence seem to be satisfactory in the midterm, although it is necessary to have at least 5 years of follow up to be able to compare it with consolidated techniques considered the gold standard in the treatment of this disease. PMID- 12564068 TI - [Periurethral injectables in the treatment of urinary incontinence caused by sphincter insufficiency]. AB - OBJECTIVE: Periurethral injectables are among the multiple therapeutic options that urologists have to correct female stress urinary incontinence currently. The philosophy of periuretrhal injectables action mechanism has always been the same, to increase urethral walls coaptation or pressure, which is pathologically diminished. METHOD/RESULTS: New surgical techniques, approaches and instruments have been appearing with time. Historically various injectable materials have been used, each one with its advantages and disadvantages. CONCLUSIONS: The role of periurethral injectables in the treatment of female stress urinary incontinence is limited. The efficacy of the procedure is low and they would have little room in the treatment of female stress urinary incontinence if it were not because of its minimally invasive condition in patients with intrinsic sphincter insufficiency, whom would require more aggressive procedures otherwise also of uncertain results, or to remain untreated. PMID- 12564069 TI - [Female stress urinary incontinence. Surgical repair with pubovaginal sling techniques]. AB - OBJECTIVES: To review the treatment of female stress urinary incontinence by new systems of tension-free urethral sling TVT type (Tension free vaginal tape) or IVS (intravaginal slingplasty), and the bone anchoring trasvaginal sling procedure Infast. METHODS: We describe the surgical techniques of the various procedures and perform a bibliographic review on the topic. RESULTS/CONCLUSIONS: The pubovaginal sling has become the gold standard in the treatment of female stress urinary incontinence, mainly if there is sphincter intrinsic dysfunction. The concept of tension free medium urethra support has been the most important contribution, that questions the classification of incontinence in types I, II and III, because the pubocervical tension free sling can correct all three. Tension free urethral sling techniques have demonstrated to be effective, minimally invasive with a low complication rate, easily reproducible, and with good continence results in the mid-term. PMID- 12564070 TI - [Obstruction following surgical repair of female stress urinary incontinence. Diagnosis and treatment]. AB - OBJECTIVES: To report our experience in the diagnosis and treatment of the lower urinary tract obstruction after urinary incontinence corrective surgery, analysing the different techniques performed, retropubic or vaginal urethrolysis. METHODS: We report a series of 14 patients with the diagnosis of obstruction after incontinence corrective surgery. They were classified in 2 groups, those who underwent retropubic procedures (5 cases) and those who underwent prolene mesh sling procedures (9 cases). We detail preoperative clinical-urodynamic parameters, and postoperative bladder outlet obstruction confirmation. Retropubic urethrolysis was performed in all patients after retropubic surgery, with the association of hysterectomy if indicated. The performance of a new sling- urethropexy was individualized. In three cases of retropubic surgery repeated urethropexy was not done. Unilateral section was performed in the sling series for all except one case of double section. A new mesh sling was performed in two cases; two cases did not undergo reoperation. Urethrolysis results were evaluated both subjectively by satisfaction degree scales and objectively by clinical urodynamic evaluation, comparing clinical data and urodynamic parameters using the proper statistical test. RESULTS: In the retropubic surgery group all patients are continent. Satisfaction degree is: very satisfied 3 patients and 2 quite satisfied. Two presented with voiding urgency not needing anticholinergic drugs, and their urodynamic data returned to normal values without post void residual. The obstructed patients in the sling group reported minimal urgency after urethrolysis in 2 cases. Two patients have stress urinary incontinence: one of them is better than before and refused to undergo a new operation; the other one, who repeated sling, developed a clinical picture of urgency-incontinence again, underwent second section and continues having stress urinary incontinence. Among 7 patients undergoing sling, 4 are very satisfied, 1 quite satisfied, 1 somewhat satisfied and 1 not at all satisfied. Voiding difficulties have disappeared in all cases; comparisons between pre and postoperative urodynamics maximum flow, detrusor pressure at maximum flow and postvoid residual show statistically significant differences. Post-urethrolysis parameters return to preoperative values. CONCLUSIONS: The immediate development of symptoms after surgical correction of urinary stress incontinence is the best diagnostic criteria for obstruction. Detrusor muscle responds to obstruction, but sometimes its response is so minimal that it is difficult to diagnose urodynamically. The knowledge of preoperative values helps to confirm the diagnosis. In any case, urodynamic parameters did not influence the success of urethrolysis. Urethrolysis is an effective operation to cure symptoms secondary to obstruction after incontinence corrective surgery. When a sling has been the procedure performed, probably it is not worth to perform a standard urethrolysis; a simple section of one of the branches seems to be enough to improve symptoms. Currently, there is not scientific evidence about the convenience or not of bladder neck-urethral re suspension after urethrolysis. The only case-scenario in which it is clearly indicated is that when there is stress incontinence in addition to obstructive symptoms. PMID- 12564071 TI - [Radical cystectomy. Bladder substitution (in women)]. AB - OBJECTIVES: To report our experience with a series of 10 patients undergoing radical cystectomy with bladder substitution. We were supported by a better knowledge of the female continence anatomical mechanisms and the demonstration of the oncological viability of the urethral remnant. METHODS: From 1994 to 2002 10 women underwent radical cystectomy with bladder substitution by means of a modified anterior pelvic exanteration; technical modifications to achieve continence preservation are based on: preservation of the distal 2/3 of urethra, pubourethral ligaments and endopelvic fascia, and limitation of lateral vaginal dissection to avoid damage to the striated sphincter innervation. To avoid the neocystocele effect the vagina is fixed to the uterosacral ligaments and to the sacral promontory. RESULTS: Bladder capacity is 332.9 +/- 35.6 ml with a flow of 17.7 ml/sec. Complete continence wax achieved in 8 patients, the remainder 2 present grade II urinary stress incontinence. All of them empty their neobladder satisfactorily; only one patient needed a bladder re-education program. One bladder fistula and diarrhoea in one patient are the complications to be highlighted. CONCLUSIONS: The results obtained with orthotopic neobladder in females, achieving a high satisfaction level and quality of life, stimulate us to continue with this technique. PMID- 12564072 TI - [Changes in the urogenital sinus]. AB - OBJECTIVES: To review the most adequate treatment for female urogenital sinus congenital malformations currently. METHODS: We performed a bibliographic search on this rare disease and review the embryological development of the female urogenital apparatus. RESULTS/CONCLUSIONS: We review the various surgical techniques from a historic point of view pointing at what time should the procedure be done. PMID- 12564073 TI - [Female urethral diverticuli]. AB - OBJECTIVES: To review the etiopathogenic, diagnostic and therapeutic criteria for female urethra diverticula. METHODS: Urethral diverticulum was diagnosed in four patients, all of them with recurring urinary tract infections and irritative voiding symptoms. The final diagnosis was established by VCUG (Voiding cystourethrography). Other procedures, more recent and more in use, are also referred. RESULTS: All patients required surgical treatment, three of them with good results and one without success. The therapeutic procedures most frequently used in the literature during the last decade are also mentioned. CONCLUSIONS: The milestone in the management of female urethral diverticula is to think on it in patients with lower urinary tract symptoms. Surgical excision when amenable obtains optimal results. PMID- 12564074 TI - [Vesicovaginal fistula]. AB - OBJECTIVES: To review the etiology, diagnosis and available therapeutic options for vesicovaginal fistula. METHODS: Bibliographic review on diagnosis and treatment of vesicovaginal fistulas. RESULTS: We review the complete diagnostic protocol for vesicovaginal fistulae, as well as the various surgical options available, all of them with high success rates. There is an increasing tendency to early repair instead of delayed. It is important to follow the principles of fistulae closure to achieve success. In cases of complex fistulae the use of interposed flaps allows to have better results. CONCLUSIONS: There are few vesicovaginal fistulae not amenable to surgical treatment; to suspect a fistula at the time of diagnosis and an early treatment with a good technique, regardless of the approach, guarantee a high success rate. PMID- 12564075 TI - [Interstitial cystitis: update on pathogenesis, diagnosis, and treatment]. AB - OBJECTIVES: Interstitial cystitis is a nosological entity of which many etiological, diagnostic and therapeutic features were unknown until recently, so that we had the objective to review new knowledge acquired during the last decade to offer an update in the disease. METHODS: A bibliographic review on interstitial cystitis is performed with special interest in national and international authors who have experience and large case series. RESULTS: Among the multiple etiopathogenical theories proposed in the past, the hypothesis to be highlighted, that is gaining strength, is the altered bladder urothelium permeability or hemato-urinary barrier alteration, with the aggression of toxic substances contained in urine in high concentrations like potassium. Epidemiologically, some authors think the disease is present in the population in a higher frequency than it is diagnosed, and it may be found in many women with the misdiagnosis of recurrent urinary tract infections. Regarding diagnosis, a new test is proposed to help clinical diagnosis: the potassium test, which consists in the endovesical instillation of this ion at high concentrations reproducing the symptoms. Finally, in the medical treatment it is demonstrated the efficacy of drug associations over monotheraphy (heparinoids, antihistaminic and antidepressant drugs), looking for the correction of the physiopathological abnormalities caused by the disease. Psicologic support and continuous information to the patient about the disease continue to be one of the mainstays of this multimodal therapy. New options like neural stimulation have appeared, although its results are not conclusive so that further studies must be done. CONCLUSIONS: New knowledge about interstitial cystitis allow us to better understand the chain of events happening in it. An early clinical diagnosis, now reinforced with a positive potassium test, is key to start medical treatment (drug combination) targeted to stop the pathogenic process, because it is more effective in the early phases of the disease. For that purpose, the Urologist must have this disease in mind. PMID- 12564076 TI - [Urinary tract infection in pregnancy and menopause]. AB - OBJECTIVES: To review the topic of urinary tract infections (UTI) during pregnancy and menopause. UTI during pregnancy and menopause have great relevance in the field of urologic infections; during pregnancy because of the particularities involved in its diagnosis and treatment and potential consequences to the fetus and mother; menopausal UTI because this group of women is numerous and represents a growing section of the general population pyramid, due to the aging of population in developed countries associated with longer life expectancies and grater demand for quality of life. METHODS AND RESULTS: We performed a bibliographic review combined with our personal experience. During pregnancy there are several functional and anatomical changes that condition not only a higher risk of UTI, but also an additional treatment difficulty due to antimicrobial pharmacokinetics alterations and potential damage to the fetus. Despite efforts to find an easy, fast and reliable test for bacteriuria detection, urine culture continues to be the first diagnostic test for its detection and follow up during pregnancy. Penicillin derivates and cephalosporins continue to be the first choice because their lack of adverse effects on either fetus or mother. Alternative options like phosphomicin and aztreonam although they show low toxicity there is need for more studies supporting their suitability for the treatment of pregnancy UTIs. Menopausal female UTI have their different features from those in younger women. Hormonal alterations derived from gonadal atrophy associate functional changes in the vaginal ecosystem, making it prone to enterobacteriaceae colonization as a first step up to the urinary tract. This associated with genitourinary tract anatomical alterations inherent t aging make UTI extraordinary prevalent in this growing segment of population. Treatment lines focus on hormonal alteration correction and proper antimicrobial prophylaxis and vaccines in a close future. CONCLUSIONS: UTIs during pregnancy and menopause have differential features that require different diagnostic and treatment approaches. PMID- 12564077 TI - Prognostic value of normal myocardial perfusion scintigraphy in patients with chest pain and left bundle branch block. AB - BACKGROUND: The purpose of this study was to assess the prognostic value of normal myocardial perfusion scintigraphy in patients with chest pain and left bundle branch block (LBBB) undergoing myocardial perfusion scintigraphy without ischemia. METHODS: Retrospectively (between February 1999 and March 2001) we assessed consecutive patients with LBBB and chest pain of suspected ischemic origin who underwent scintigraphy in our institution with tetrofosmin (technetium 99m) and thallium-201 SPECT one-day dipyridamole or adenosine stress-rest protocol and no evidence of ischemia. Of a total of 61 patients with a mean age of 67.6 +/- 9.7 years, 23 (37.7%) were male and 38 (62.3%) female. Hospital admission or death from cardiac cause, suspected ischemic pain and infarction were classified as hard events. Outcome was defined as survival free of events. We used Cox regression to assess the univariate association between outcome and the variables gender, ejection fraction < 45%, diabetes, hypertension, smoking, and LDL-cholesterol > 130 in a period of one month around the date of scintigraphy, and body mass index (BMI) > or = 25; the Kaplan-Meier test to assess the relation of prognosis to hard events; and the chi-square test to assess the distribution between genders. RESULTS: The mean follow-up was 14.2 +/- 7.0 months. 19.7% were smokers, 70.5% had a history of hypertension, 19.7% were diabetics, 64.8% had a BMI > or = 25, 53.8% had LDL-cholesterol > 130 mg/dl and 36.1% had an ejection fraction < 45%. Nine patients (14.8%) had a hard event (5 hospital admissions and 4 cardiac deaths). We found an association between diabetes and outcome (hard event relative risk 5.7; 95 percent confidence interval 1.53 to 21.37; p = 0.009) but not with other variables. CONCLUSIONS: Myocardial scintigraphy was shown to be a useful prognostic tool in our patients with LBBB and suspected coronary artery disease, and no evidence of ischemia on scintigraphy. 14.8% patients had a hard event in a mean follow-up of 14.2 +/- 7.0 months. Patients with diabetes had a 5.7 times increased relative risk for a hard event. PMID- 12564078 TI - Normal myocardial perfusion scintigraphy and left bundle branch block: good prognosis in all patients? PMID- 12564079 TI - Long-term prognostic value of the hemorheological profile in transmural myocardial infarction survivors: 60-month clinical follow-up. AB - Previous reports have shown several hemorheological and hemostatic abnormalities in acute coronary syndrome survivors. Some of these abnormalities were related to cardiovascular events during a 24-month follow-up. The aim of the present work is to evaluate, in transmural myocardial infarction survivors, the long-term (60 months) prognostic value of the biohemorheological profile determined at hospital discharge. Sixty-four patients (59 men), mean age of 58 +/- 12.0 years, transmural myocardial infarction survivors, were prospectively studied for 60 months (32.0 +/- 17 months, median 33 months). The following cardiovascular events (CVE) were analyzed: death, non-fatal infarction, unstable angina, and stroke. Twenty-nine patients had a CVE (nine died). The following parameters were determined at hospital discharge: plasma viscosity, whole blood viscosity, erythrocyte membrane fluidity, erythrocyte aggregation, protein C, plasminogen inhibitor type I (PAI-1), leukocyte count and elastase. The quartiles were determined for each parameter, grouping patients according to these values. STATISTICS: Group-t-test, Kaplan-Meier survival curve (with log rank test), and Cox logistic regression. RESULTS: 1) Leukocyte count (p < 0.01), protein C activity (p < 0.05) and erythrocyte membrane fluidity (p < 0.05) were predictors of the CVE curve; 2) The higher the value of the leukocyte count quartile, the higher the risk for a CVE (p < 0.05). Patients with a leukocyte count above the median had 4 times more risk for a CVE; 3) The lower the protein C activity, the higher the risk for a CVE. Those with protein C activity lower than the lowest quartile had double the risk; 4) The higher the membrane polarization value (membrane rigidity), the higher the risk of a CVE; 5) By multivariate analysis the 3 parameters were independent predictors of a CVE. CONCLUSION: In the present group of transmural myocardial infarction survivors a close relationship was established between hemorheologic, hemostatic and inflammatory factors and the cardiovascular events curve during long-term follow-up. PMID- 12564080 TI - Evaluation of bilateral pulmonary vein isolation for the treatment of atrial fibrillation: value of postoperative electrophysiological study. AB - OBJECTIVE: To evaluate electrophysiological changes after bilateral pulmonary vein isolation in patients undergoing mitral valve surgery and to relate the clinical outcome at 1-year follow-up with the results of the postoperative electrophysiological study. METHODS: Prospective study of patients with atrial fibrillation undergoing bilateral pulmonary vein isolation using radiofrequency energy and concomitant mitral valve surgery. Pulmonary vein isolation was performed with a heptapolar catheter. Each set of up to 7 simultaneous applications aimed at a maximal duration of one minute and a maximal preset temperature of 70 degrees C. Energy delivery (< 150 watts) was controlled by thermosensors located on each electrode. Before hospital discharge, all patients presenting sinus rhythm underwent an electrophysiological study, using epicardial pacing wires placed during surgery. Isolated areas were compared to the non isolated left atrium with respect to electrogram amplitudes and atrial capture thresholds. At one-year follow up, cardiac rhythm was assessed in all patients and correlated with the results of the postoperative electrophysioloical study. RESULTS: The study population consisted of 20 patients with mitral valve disease (mean age 59 +/- 11 years), chronic atrial fibrillation present for 0.5 to 18 years and dilated left atria (55 +/- 11 mm assessed by M-mode echocardiography). The radiofrequency procedure added, on average, 7 minutes (3 to 17) to the duration of the mitral valve surgery. No patient died or suffered surgical complications. Eleven patients presented sinus rhythm before hospital discharge. The electrogram amplitude inside the isolated zones was less than 25% of that in nonisolated areas (0.3 +/- 0.2 mV versus 2.1 +/- 1.7 mV, p = 0.002). Atrial capture thresholds were significantly higher for the isolated areas (13.5 +/- 9.3 mA versus 8.5 +/- 4.0 mA; p < 0.05). At one-year follow-up, nine patients maintained atrial fibrillation, nine remained in sinus rhythm and two lost sinus rhythm to atrial fibrillation. In both these patients, the reduction in electrogram amplitude inside the isolated zones was similar to that of the remaining patients discharged in sinus rhythm. However, postoperative electrophysiological studies in these patients showed that atrial capture thresholds for isolated areas had not increased significantly (8 mA and 6 mA, respectively). CONCLUSION: Rhythm at discharge after bilateral pulmonary vein isolation tends to remain at one-year follow up. Radiofrequency-induced modifications in electrogram amplitudes may be less important than increases in pacing thresholds of isolated zones with respect to maintenance of sinus rhythm. PMID- 12564081 TI - Evaluation of biventricular function in the rat: a new experimental model. AB - The use of small animals in cardiovascular research has increased over recent years. This might be a limitation when evaluation of biventricular function is required. Although evaluation of left ventricular (LV) pressure and volume is already possible in small animals, concomitant evaluation of right ventricle function has been limited to large animals. The study describes a new model to assess pressures and dimensions of both ventricles simultaneously in the adult rat. Adult Wistar rats (n = 12), weighing 372 +/- 16 g, were anesthetized with pentobarbital (60 mg/kg, i.p.) and ventilated through a tracheostomy (60 cpm, 1 ml/100 g). Under a dissecting microscope (6x) the right jugular vein was catheterized. After sternotomy and pericardiotomy, three crystals were placed along the major cardiac transverse diameter: in the right subendocardium of the interventricular septum and on the epicardial surfaces of the RV and LV free walls. In addition, two high-fidelity catheters were introduced through the apex into the RV (2F, Millar) and LV (3F, Millar) cavities. This allowed the measurement of all parameters derived from pressure and dimension curves of the RV and LV, including pressure-dimension loops. This study describes, for the first time, a model that allows simultaneous evaluation of biventricular pressure and dimensions in an animal model as small as an adult rat. This model opens up new perspectives for the establishment of correlations between molecular biology and hemodynamic data in both ventricles, which is particularly important as more differences between the two ventricles are being found. PMID- 12564082 TI - Sudden death and variant angina. AB - Variant angina is defined by chest pain occurring at rest associated with transitory ST segment elevation on ECG, and is caused by a spasm of a coronary artery. Frequently, variant angina is associated with atherosclerotic coronary obstruction and patients with normal coronary arteries are rare. Patients with variant angina and normal coronary arteries have good prognosis, and the development of ventricular arrhythmias or sudden death is rare. The authors present two cases of sudden cardiac death in patients with variant angina and normal coronary arteries. PMID- 12564083 TI - Radiofrequency ablation in a patient with atrioventricular reentrant tachycardia and atrioventricular nodal reentrant tachycardia with 2:1 atrioventricular block. AB - The authors report the case of 15-year-old girl with a history of palpitations and shortness of breath during exercise. The electrocardiogram showed ventricular preexcitation suggesting a Wolff-Parkinson-White syndrome with a posteroseptal accessory pathway. During the electrophysiological study a left posterospetal accessory pathway was identified and an orthodromic atrioventricular reentry tachycardia was reproducibly induced (cycle length 400 ms). After disappearance of the ventricular pre-excitation with radiofrequency ablation, a dual physiology of atrioventricular node condution was documented and a slow-fast atrioventricular nodal reentrant tachycardia was repeatedly induced. Upon induction, this tachycardia presented a proximal atrioventricular block with 2:1 condution converted to 1:1 condution with overdrive pacing from the proximal coronary sinus (cycle length 270 ms). Radiofrequency ablation of the slow pathway was performed with success. We discuss the need to suspect and seek different arrhythmogenic substracts of tachycardia in a single patient, the electrophysiologic conditions that could explain the inducibility of different arrhythmias in this case, and the controversy regarding ablation of more than one reentry circuit in a single procedure. PMID- 12564084 TI - C-reactive protein: a new golden marker of cardiovascular risk. AB - The need for new cardiovascular risk factors, due to the limitations of traditional factors, is pointed out. Recent views regarding the role of inflammation in the evolution of the atherosclerotic process, namely in the formation of the atheroma plaque, its progression and rupture, and initiation of acute coronary events, are briefly summarized. These events point to the possibility that inflammatory markers could constitute new markers of cardiovascular risk. Amongst them, C-reactive protein (CRP)--particularly high sensitivity CRP (hsCRP)--has been found to be a 'golden marker' in this field. Data are presented indicating that CRP, rather than being merely an epiphenomenon of inflammation, in fact constitutes a pathogenic agent of the atherosclerotic process. The importance of CRP elevation in acute coronary events, in revascularization interventions, in other situations of cardiovascular risk and even in apparently healthy individuals is outlined. The performance of CRP is then compared with that of traditional risk factors. Finally, the need for studies to discover whether a decrease in CRP levels, as obtained with several pharmacological agents, is accompanied by a simultaneous reduction of cardiovascular risk is emphasized. PMID- 12564085 TI - Cardiac disease outcomes in clinical trials. AB - Randomized controlled trials (RCTs) are the gold standard for determining causality in medicine. To be considered valid, the efficacy/effectiveness of a new drug must be tested and proved through this type of scientific study. This type of trial will disclose the drug's risk profile as well as the treatment effect magnitude. The proper design, development, and analysis and presentation of results from an RCT is based on a group of well-defined methodological rules, compliance with which assures the trial's internal validity, the relative and absolute importance of the results and its applicability to populations of patients different from those included in the study sample (external validity). Among the structural and methodological components of a clinical trial- randomization, allocation concealment, confounding, similarity of study groups, measures of efficacy, statistical analysis, etc.--disease markers (endpoints, outcomes) are especially important. In the end, what an RCT is good for is to detect changes in the disease process with therapy (or preventive measures), and these changes are defined beforehand based on specific measurements--disease markers. In this paper we will present general principles for the definition of disease markers, their problems and practical use. Caution should be exercised, as this is an area of clinical epidemiology that is somewhat complex, controversial and ill-defined. PMID- 12564086 TI - Percutaneous retrieval of central venous catheter fragment in a premature newborn. PMID- 12564087 TI - Cardiac mass in a patient with chronic lymphocytic leukemia. PMID- 12564088 TI - [Early results of mitral valve surgery]. AB - The authors submit a retrospective group of 245 patients operated in the course of 6 years at the Second Surgical Clinic of Cardiovascular Surgery of the General Faculty Hospital and First Medical Faculty Charles University on account of mitral valve disease. The early results of mitral valve prostheses are comparable or in some combined operations better than reported in world statistics. In the etiology the authors found a relative decrease of rheumatic defects and higher incidence of ischaemic affections. The general mortality in mitral valve prostheses was 7.1%, combined operations incl. acute ones being predominant. In mitral plastic operations the authors lost 4 patients, i.e. 19%. In all instances acute combined operations were involved and the result is consistent with the predicted mortality according to the Merged Cardiac Registry. The mean hospitalization period was 7.52 days. A postoperative QIM was recorded in one patient, i.e. in 0.46%, revision after mediastinitis also in one patient (0.41%) and revision after haemorrhage in 14 patients, i.e. in 5.7%. Neurological complications were recorded in 10 patients i.e. 4%. The work emphasizes the trend of improving results in this important area of cardiosurgery and indicates further possibilities and ways of development in the treatment of mitral valve diseases. PMID- 12564089 TI - [Treatment of pseudoaneurysms with injection of thrombin]. AB - OBJECTIVES: To introduce and assess the efficacy and risks associated with a new treatment method of post-catheterization pseudoaneurysms. METHODS: Thrombin injection was indicated in patients with failure of compression therapy during 24 hours. Patients with a cavity volume < 1 cm3, with a pseudoaneurysm causing compression or haemorrhagic shock were excluded. A needle is introduced under local anaesthesia with ultrasound guidance, its tip is placed thoroughly in the centre of the cavity. Subsequently, under colour-flow mapping, thrombin solution with concentration of 1000 IU/ml is cautiously injected, which results in instantaneous thrombosis of the pseudoaneurysm cavity. RESULTS: Thrombin injection was used in 17 patients during a period of 12 months. 12 of them were women, 6 patients with valvular disease, 2 after intervention and 5 anticoagulated. The method was successful in 16 patients. In 5 of them, a small residual cavity persisted, which was not suitable for re-injection, and was closed completely after a few minutes of compression by a ultrasound transducer. There were no thrombotic complications. CONCLUSION: Although compression remains the method of choice, thrombin injection is a highly effective, quick and, with necessary precautions, safe alternative to surgical treatment of pseudoaneurysms, mainly in anticoagulated patients. PMID- 12564090 TI - [Endovascular treatment of atherosclerotic stenoses of the subrenal aorta]. AB - The authors present results of endovascular treatment of subrenal aortic atherosclerotic stenoses. Between 1996 and 2001 they treated 9 patients (5 men, 4 women, mean age 57.4 years). Aortic stenoses were caused by an atherosclerotic process in all cases. In addition, in three patients iliac arteries were affected by sclerotic lesions. The endovascular procedure consists of predilatation performed by a small calibre angioplasty balloon catheter and implantation of a stent. Three times a balloon expandable stainless steel stent was implanted and self-expandible nitinol stents were used in the six cases. The primary technical success rate was 100%, no major complications were observed. One distal embolization was successfully resolved by intraarterial thrombolysis. Only one stent restenosis occurred after 36 months. It was successfully treated by implantation of a longer stent. That means that the primary patency of stented subrenal aorta during a mean follow-up period of 19 months was 87.5%. The claudication interval improved in six patients and three patients were symptom free. Endovascular therapy can be recommended for short sclerotic stenoses of the infrarenal aorta. This miniinvasive technique has a high technical and clinical success rate, favourable long-term patency and low complication rate. PMID- 12564091 TI - [Surgical treatment of hyperparathyroidism--personal experience]. AB - The authors submit their own experience with the surgical treatment of all types of hyperparathyroidism (HPT). Since 1992 till the end of April 2002 in the Institute of Clinical and Experimental Medicine 151 operations were performed in 132 patients, in particular secondary HPT (69.8%). Among surgical operations partial parathyroidectomy (PTE) predominated, as a rule without peroperative implantation of parts of the removed parathyroid glands(PT)(64.8%). The high ratio of this type of operations was due among others to a recent increase of tertiary posttransplantation HPT. Advanced secondary HPT of haemodialyzed patients was treated by total PTE with implantation of PT tissue. The incidence of surgical complications was low (7.1%) and insignificant from the aspect of the long-term postoperative condition. Persisting or relapsing HPT was revised in 15 patients (in 12.6% of all operations). PMID- 12564092 TI - [Infiltration of local anesthetics into the thyroid gland capsule for surgery and the postoperative period]. AB - In a prospective randomized study the hypothesis was tested whether infiltration of the thyroid capsule by a local anaesthetic will reduce the haemodynamic response to surgical trauma, consumption of anaesthetics and opioids during surgery and will shorten the time of arousal. A total of 64 patients indicated for planned goitre surgery were divided at random into a control group (C, n = 32) and experimental group (LA, n = 32). The preoperative medication and anesthesia did not differ in the two groups. In group C into the thyroid capsule a maximum of 40 ml saline was administered, in LA the same volume of 0.5% trimecain. A highly significant difference was found (p < 0.001) in the incidence of hypertension during surgery (21 C vs. 5 LA) and the need of further pharmacological interventions (21 vs. 8). In the control group was a higher consumption (p < 0.95) of the opioid phentanyl (167.5 +/- 111 micrograms vs. 125 +/- 93.5 micrograms), a trend of longer arousal and the need to antagonize the effect of opioid (p < 0.1). The substitution of saline by a local anaesthetic for infiltration of the thyroid capsule is a safe and simple method leading to a reduction of cardiovascular complications during surgery. PMID- 12564093 TI - [Laparoscopic splenectomy--personal experience]. AB - The authors present an account on their own experience with laparoscopic splenectomy which is used at the First Surgical Clinic, First Medical Faculty and General Faculty Hospital in Prague since 1996. In 1996 to 2002 the authors performed at the Surgical Clinic a total of 66 splenectomies. This number included on account of injury of the spleen, tumour or cyst 12 splenectomies by the open route. The remaining ones were indicated for haematological reasons in collaboration with the haematological department of the First Medical Clinic of the First Medical Faculty and General Faculty Hospital in Prague and the Institute of Haematology and Blood Transfusion in Prague. 23 splenectomies for haematological reasons were made by the classical open route. In 31 patients the splenectomy was performed by the laparoscopic route. In all haematological reasons were involved. In the group operated at the First Surgical Clinic LSE was indicated because of ITP 15x, for spherocytosis 8x, for haemolytic anaemia 7x, for eliptocytosis 1x. In eight patients at the same time laparoscopic cholecystectomy was performed. In the group subjected to classical splenectomy infection in the surgical wound was recorded in 11%, re-operations on account of a suppurative complications in the abdominal cavity were made in 13% and on account of haemorrhage in 11%. In the group of laparoscopic splenectomies the authors did not record infection at the site of the inserted trocars, there were no suppurative complications in the abdominal cavity. In four laparoscopic operations the authors converted the operation on account of haemorrhage four times (11%), once on account of major adiposity of the omentum, in five postoperative revision on account of haemorrhage was necessary (16%), incl. one case of profuse haemorrhage. Therefore the authors sought a way how to prevent haemorrhage. Based on their own experience they recommend to apply clamps to the trunk of the lienal artery. The time of operation was reduced by the application of clamps to the lienal artery to 60-80 mins. and the peroperative blood losses dropped to 20-30 ml. The morbidity declined and the patients are discharged into domiciliary care on the 4th day after operation, to be on the safe side. No late complications of the operation were recorded. PMID- 12564094 TI - [Tumor biopsy--a trivial matter or a basic procedure?]. AB - The surgeon is frequently faced with the task of bioptic sampling of tissues from patients with tumourous diseases. The importance of such frequently minor procedures is incorrectly underrated. The importance of biopsy is in the foreground in particular at present at a time of individualization of treatment of malignant tumours, based on the development of new diagnostic-therapeutic methods. The bioptic specimen is the fundamental link in the basic decision- benignity or malignity--and also a unique source for assessment of prognostic and predictive factors on the basis of which we select the optimal therapeutic procedure. Contrary to current histopathological examination where the principles of correct collection of tissues specimens are generally known, in recent years the importance of molecular examinations is increasing where the surgeon must respect certain different principles to make the sampling successful. The authors working in a department of surgical oncology along with authors from specialized laboratories formulate rules of correct implementation of biopsies and transport of biological material in conjunction with recent laboratory methods, and based on examples of their own practice, they demonstrate how the initial approach of the surgeon can influence in a decisive way the correct diagnosis and therapeutic procedure in oncological patients. PMID- 12564095 TI - [Tumors of peripheral nerves of extremities and the brachial plexus and their surgical treatment]. AB - BACKGROUND: The author presents the results of 32 operations in 30 patients to whom in the course of 11 years, from 1990-2001 by the microtechnique 32 tumours of peripheral nerves of the extremities and the brachial plexus were removed. PATIENTS AND METHODS: In 26 patients 28 tumours of the peripheral nerves of the extremities were removed and in 4 patients 4 tumours of the brachial plexus were removed. Three of the tumours of the brachial plexus spread intraspinally. 31 tumours of the nervous sheaths included 19 schwannomas and 12 neurofibromas. Of 12 neurofibromas six were found in four patients with Recklinghausen's disease (VRD). One patient had a tumour of non-neural elements--a lipoma. The authors did not encounter malignant tumours of nervous sheaths, benign or metastatic malignant ones which compressed or invaded the nerve or plexus. The results were evaluated according to Donner's classification of motor and sensitive functions. RESULTS: Motor functions improved but remained unaltered in 17 (89%) patients with schwannomas and in 5 (83.3%) patients with pain in the area of the affected nerve either complete or partial mitigation of symptoms occurred. In neurofibromas in 3 (50%) patients motor functions improved or remained unaltered and all patients had a partial or complete regression of painful syndromes. In 4 patients with VRD 6 tumours of the peripheral nerves of the extremities were removed, in all the motor functions improved or remained unalterted and in all a partial or complete regression of pain syndromes was recorded. The results were more favourable in smaller, more distant tumours of the nerve sheaths. CONCLUSION: Patients should be operated in departments concerned with problems of peripheral nerves. PMID- 12564096 TI - [Acute abdomen after cardiac surgery]. AB - Acute abdomen after cardiosurgery accounts for a minor group of complications, however, they are associated with a high mortality and morbidity. In 2001 in the authors department 940 cardiac operations were performed. In four patients (0.43%) during the early postoperative period acute abdomen developed which called for urgent surgical intervention. Two patients died. Only early diagnosis and subsequent urgent laparotomy can have a positive impact on the prognosis of this group of patients. PMID- 12564097 TI - [Image of the month. Pulmonary malacoplakia due to Rhodococcus equi in a patient with AIDS]. PMID- 12564098 TI - [Clinical case of the month. Constipation and abdominal mass syndrome in a 51 year-old patient]. AB - A 51-year old patient consults for abdominal swelling and persistent constipation. Clinical exploration shows the presence of a left iliac fossa tumor corresponding to a papillary serous adenocarcinoma of the fallopian tube after macroscopic and microscopic examination. The diagnostic and therapeutic problems caused by this rare gynecologic tumor are discussed. PMID- 12564099 TI - [Cannabis, status in 2001-2002]. AB - Even though its use becomes common, cannabis remains a narcotic, the risks of which are nowadays better known. Its therapeutic value still needs to be scientifically demonstrated. PMID- 12564100 TI - [Quality of life assessment in clinical trials: a holistic approach to health]. AB - The concept of quality of life has been widely developed over the last two decades. As they provide subjective assessment of health, quality of life measures may be broadly and relevantly applied in many areas. Numerous measures are available today to perform discriminative, evaluative or analytical studies in the epidemiological, the therapeutic or the health economic fields. Such measures are increasingly used and may be added to clinical, biological or radiographical criteria to assess medical benefits and health outcomes in order to strengthen decisional choice in health. PMID- 12564101 TI - [Evaluation criteria for acute pancreatitis. Recent data from the literature]. AB - This paper related to the diagnosis and etiological factors of acute pancreatitis (AP). 20% of all cases of AP will develop local and/or systemic complications; infections of the lesions play an important role. To predict severity and outcome, the value of clinical examination, current biological parameters, new markers and computed tomography is discussed. PMID- 12564102 TI - [Autoimmune polyendocrine syndrome (APS)]. AB - As a complement to basic research, thorough clinical investigation of rare diseases may provide fundamental elements which improve our understanding of still obscure pathophysiologic mechanisms. This is the case with immunoendocrinopathy syndromes. Since Addison's pioneer observations in the 19th century, physicians have known that some individuals and their families may be affected by several spontaneous endocrine insufficiencies that are associated with autoimmune extra-endocrine processes. APS-I or APECED syndrome appears in children firstly affected by recurrent muco-cutaneous candidiasis and hypoparathyroidism, followed by adrenocortical insufficiency and by other autoimmune processes. APS-I is a monogenic disorder resulting from one mutation in the AIRE gene. The protein encoded by AIRE is a nuclear transcription factor the precise target of which is still not known. AIRE is mainly expressed by cells playing a crucial role in the establishment of central T cell self-tolerance (medullary epithelium, macrophages and dendritic cells of the thymus). APS-I must be considered in children affected with recurrent candidiasis without any sign of primary immune deficiency. Scientific investigation of the biological nuclear events controlled by AIRE has to be pursued. Undoubtedly, their deciphering will increase our knowledge of the mechanisms responsible for the establishment of central T cell self-tolerance and will open novel strategies for managing many autoimmune diseases. APS-II is a more common syndrome characterized by adrenocortical insufficiency spontaneously occurring in non tuberculous adults and associated with autoimmune thyroiditis and/or type 1 diabetes. Contrary to APS-I, APS-II is linked to genetic loci of the major histocompatibility complex. There is no adrenal insufficiency in APS-III which includes autoimmune thyroiditis, type 1 diabetes, and other autoimmune extra-endocrine processes (like pernicious anemia and vitiligo). PMID- 12564103 TI - [Nosologic overview of the spectrum of cutaneous histiocytoses. The more things change, the more they stay the same]. AB - Histiocytes belong to distinct groups of cells. Similarly, histiocytoses represent some pathologic expressions of the various histiocyte lineages recognized by their distinct phenotypes. The recognition of four major histiocytosis types relies on the distinction between monocytes-macrophages, dermal dendrocytes, Langerhans cells and the so-called indeterminate cells. Each of these groups exhibits a distinctive clinical and histological presentation. The risk of internal involvement, the prognosis and therapy are different as well. The diagnostic mainly relies on the clinico-pathologic assessment, and more particularly on the combination of histological, immunohistochemical and ultrastructural examinations. PMID- 12564104 TI - [Therapeutic advances in 2002 (1)]. AB - The most important drugs registered and/or launched in Belgium during the last year in the various disciplines of internal medicine will be briefly described. The main characteristics of each molecule and its modalities of appropriate use in clinical practice will be emphasized. For all these molecules, both the efficacy and tolerance have been proven in randomised clinical trials according to the rules of Evidence-Based Medicine. PMID- 12564105 TI - [How I explore...a gynecologic pathology. Part 1: clinical history and examination]. AB - After major technical advances in obstetrics and gynecology, one can unfortunately observe a decreased quality of the clinical history and gynecological examination, and of usual paraclinical tests. However, outpatient gynecological examination is the main component of a precise diagnosis and of possible medical or surgical treatments. In this article, we review the successive steps of outpatient clinic examination, with emphasis on the important parts of clinical examination. The complementary explorations will be detailed in the second part of this article, in a future issue of the Revue Medicale de Liege. The pathologies will not be exhaustively reviewed in this manuscript; only the diagnostic side of clinical examination will be detailed. PMID- 12564106 TI - [Medication of the month. The combination of budesonide 160 micrograms/formoterol 4.5 micrograms (Symbicort TH)]. AB - The combination of budesonide 160 micrograms/formoterol 4.5 micrograms (Symbicort TH) in the same dry powder inhaler (turbohaler) results in a drug that possesses powerful bronchial anti-inflammatory, bronchoprotective and bronchodilating activities. Symbicort TH is registered as a maintenance treatment for asthma from the age of 12 years. This drug is indicated in the treatment of moderate and severe persistent asthma as well as in asthma which remains mild persistent despite the regular use of low doses of inhaled corticoids alone. Symbicort TH rapidly improves asthma control by reducing symptoms, relief medication consumption and by improving lung function. Importantly, when taken regularly over a prolonged period, the drug reduces the number of mild and severe exacerbations. The dose of Symbicort TH is adjustable according to the disease severity and generally fluctuates between 2 x 1/24 h and 4 x 2/24 h. Symbicort TH is a simple and a efficient treatment likely to improve the compliance of asthmatics to their treatment in real life. PMID- 12564107 TI - [Follow-up studies of Hokkaido residents with Kawasaki disease]. AB - OBJECTIVES: Sixteen national surveys of Kawasaki disease in Japan from 1970 to 2000 have identified a total of 169,117 patients with Kawasaki disease. Based on that figure, 8,460 residents of Hokkaido probably have a history of Kawasaki disease. It is also estimated that almost 270 Hokkaido residents would have Kawasaki disease-related coronary artery disease. We underwent follow-up studies of Hokkaido residents > or = 15 years with Kawasaki disease. METHODS: We mailed a questionnaire to the departments of internal medicine, cardiology or cardiovascular surgery inquiring about the health status of patients with Kawasaki disease at 451 hospitals with 20 or more beds in Hokkaido. RESULTS: We obtained replies from 185 hospitals (41.0%). Only 11 hospitals(5.9%) reported experience of patients with Kawasaki disease(with or without coronary artery disease) at hospital follow-up. Detailed patient histories for 60 patients from 7 hospitals were obtained: Twenty patients had Kawasaki disease complicated with coronary artery disease, whereas 40 patients had a history of Kawasaki disease and no present coronary artery disease. Thirty-seven patients without coronary artery disease were followed up at one hospital. The 60 patients were aged from 15 to 36 years. Thirty-nine patients(65%) were in the 15 to 20 year age bracket. Coronary aneurysms were recognized in 25 patients(24 males and 1 female) with Kawasaki disease(41.7%) at the onset of the disease. Twenty patients had an aneurysm(s) in the left main trunk and/or the left anterior descending artery, and 13 patients in the right coronary artery. There was a history of myocardial infarction in four patients (6.7%) and nine patients(15.0%) still suffered from angina pectoris. Aortocoronary bypass surgery was performed in one patient, whereas two patients required percutaneous coronary intervention. CONCLUSIONS: Our study suggests that the majority of Hokkaido residents with Kawasaki disease(with or without coronary artery disease) are not being followed up at area hospitals. The characteristics of patients who were followed up included age < or = 20 years, male sex and the presence of severe coronary artery disease other than circumflex involvement. PMID- 12564108 TI - [Outcome of aortic arch surgery in patients aged 70 years or older: axillary artery cannulation and selective cerebral perfusion supports]. AB - OBJECTIVES: Axillary artery cannulation, selective cerebral perfusion and replacement of the ascending and arch aorta with an elephant trunk were evaluated to reduce cerebral complications in aortic arch surgery in patients with aortic aneurysm or aortic dissection involving the aortic arch. METHODS AND RESULTS: A total of 45 patients(18 with acute A type aortic dissection and 27 with chronic aortic aneurysm involving the aortic arch) aged 70-92 (mean age 74) years underwent total aortic arch replacement from March 1996 to May 2002. There were three operative deaths in patients with acute A type aortic dissection caused by massive cerebral infarction, bleeding and myocardial infarction, and one hospital death of sepsis. Overall in-hospital mortality was 8.9%(16.7% in A type dissection and 3.7% in chronic aneurysm). Operative complications included mediastinitis in four patients(9%), left recurrent laryngeal nerve palsy in eight(18%), and cerebral infarction in four(9%). Three of the patients with cerebral infarction had associated dissection-related cerebral ischemia before surgery. One patient died, and two needed a walking stick. Twelve of 18 patients(67%) with acute A type aortic dissection and 26 of 27 (96%) with chronic aortic aneurysm were discharged on foot. CONCLUSIONS: Axillary artery cannulation, selective cerebral perfusion and replacement of the ascending and arch aorta with an elephant trunk provided satisfactory operative results in elderly patients aged 70 years or older, especially in patients with chronic aortic aneurysm involving the aortic arch. PMID- 12564109 TI - [Is blood squeezed out from the microcirculation soon after coronary occlusion?: real time myocardial contrast echocardiographic study]. AB - BACKGROUND AND OBJECTIVES: The fate of blood trapped inside the myocardial microvessels after complete occlusion of the coronary artery has not been elucidated. Blood may be squeezed out by residual contraction even after coronary occlusion. The microcirculation was examined soon after coronary occlusion by real-time contrast echocardiography. METHODS: Myocardial opacification in the short-axis view was examined during infusion of FS69 (Optison) using the Coherent Contrast Imaging method(SIEMENS Sequoia 512) in 17 open chest dogs. The left circumflex coronary artery was completely ligated for 15 sec and the video intensity (256 gray level) of the risk area was measured at end-systole of every 3 beats before and after ligation. Temporal deterioration of the wall motion was evaluated, defined as normokinesis, mild hypokinesis and severe hypokinesis, and akinesis. Changes in the video intensity during each stage of wall motion abnormalities were examined. RESULTS: The video intensity of the risk area decreased significantly after coronary occlusion. Wall motion of the risk area deteriorated gradually after ligation, in which mild hypokinesis, severe hypokinesis and akinesis developed consecutively at 2.8 +/- 0.5, 5.4 +/- 0.5 and 7.8 +/- 0.8 sec after coronary ligation, respectively. The changes in video intensity during each stage of the wall motion abnormalities were small and not statistically significant, irrespective of severity of wall motion abnormalities. CONCLUSIONS: These findings suggest that the blood remains in the vascular bed immediately after the coronary occlusion and is not squeezed out even if wall motion of the risk area continues. PMID- 12564110 TI - [Cardiomyopathy showing progression from diffuse left ventricular hypertrophy to dilated phase associated with mitochondrial DNA point mutation A3243G: A case report]. AB - A 44-year-old man was admitted to our hospital because of congestive heart failure. He had various symptoms caused by insulin-dependent diabetes mellitus, sensorineural deafness, Wolff-Parkinson-White syndrome and cardiomyopathy associated with mitochondrial DNA point mutation A3243G. Echocardiography had showed symmetrical hypertrophy of the left ventricular wall and normal cardiac function (ejection fraction 55%) at age 32 years. However, echocardiography showed cardiac transformation, consisting of posterior wall thinning and significantly reduced cardiac function (ejection fraction 11%), at age 44 years. Electrocardiography showed lowered R-wave in the chest leads and QRS widening. Both lactic acid and pyruvate serum levels were increased. Mitochondrial respiratory enzyme analysis in gastrocnemius muscle tissue indicated a partial deficiency of rotenone-sensitive NADH cytochrome C reductase. He was discharged from our hospital, and medically treated with coenzyme Q10(30 mg/day). He had no progression of cardiomyopathy or congestive heart failure. However, he suddenly died of lactic acidosis at age 47 years. PMID- 12564111 TI - [Large hiatus hernia compressing the heart and impairing the respiratory function: a case report]. AB - A 76-year-old female presented with a large hiatus hernia with intrathoracic stomach manifesting as severe exertion dyspnea. She had no cardiac or pulmonary disease, and neither anemia nor cyanosis, but respiratory function was mildly impaired. Chest roentogenography showed a large abnormal shadow overlapping the lower half of the heart. Transthoracic echocardiography demonstrated a mass compressing the left atrium and extending to the posterior part of the left ventricle, but the actual cause of the mass was not clear. Cross-sectional spiral computed tomography(CT) revealed a large hiatus hernia with intrathoracic stomach located just behind the left atrium with resultant mild anterior shift of the whole heart. Moreover, three-dimensional curved reformation CT suggested that the intrathoracic stomach was located in the upside-down position, which was confirmed by subsequent gastroesophagography. She experienced gradual progression of exertion dyspnea during the following 3 months. Follow-up CT revealed no significant increase of left atrial compression, but subsequent spirometric study showed increased impairment of respiratory function. Surgical repair for the hiatus hernia was successfully performed, and eventually achieved resolution of the symptoms. The cause of exertion dyspnea was probably cardiac compression and impaired respiratory function. The therapeutic strategy of surgical repair is recommended in elderly patients with hiatus hernia complicated with cardiac compression and respiratory impairment. PMID- 12564112 TI - [Myocardial regeneration therapy for heart failure: hepatocyte growth factor enhances the effect of cellular cardiomyoplasty]. AB - BACKGROUND: We hypothesized that transfection of the gene for human hepatocyte growth factor(hHGF) combined with cellular cardiomyoplasty might regenerate the impaired myocardium. METHODS AND RESULTS: We used a ligation model of proximal left anterior descending coronary artery of Lewis rats. Two weeks after left anterior descending coronary artery ligation, three different treatments were conducted; 1) neonatal rat cardiomyocytes group(10(6) cells, T group, n = 11), 2) HVJ-liposomes bearing the hHGF gene group (H group, n = 10), and 3) combined(T-H group, n = 10). The injection site was the scar area of myocardial infarction. For control, culture medium was injected (C group, n = 13). Echocardiography demonstrated that cardiac performance was significantly ameliorated in the T-H group 4 and 8 weeks after injection. Contrast echocardiography also showed a marked increase in myocardial perfusion in the T-H group but not in the other groups. In the T-H group, neovascularization and a marked reduction of fibrosis were observed histologically. In an immunohistochemical study, strong staining for beta 1-integrin, alpha- and beta-dystroglycan were found principally in the basement membrane of myocytes in the T-H group 8 weeks after transplantation, although there was weak immunoreactivity in the T group. CONCLUSIONS: hHGF gene transfection enhanced the cellular cardiomyoplasty possibly by stimulating angiogenesis, restoring the impaired extracellular matrix, and promoting the integration of the dissociated grafted myocytes. The combined effects might have lead to the improved cardiac performance. Thus, combined therapy may be a promising strategy for the treatment of heart failure caused by myocardial infarction. PMID- 12564113 TI - [Endothelial cell senescence in human atherosclerosis: role of telomeres in endothelial dysfunction]. AB - BACKGROUND: The functional changes associated with cellular senescence may be involved in human aging and age-related vascular disorders. We have shown the important role of telomeres and telomerase in vascular cell senescence in vitro. Progressive telomere shortening in vivo has been observed in the regions susceptible to atherosclerosis, implicating its contributions to atherogenesis. However, whether senescent vascular cells are present in the vascularture and contribute to the pathogenesis of atherosclerosis remains unclear. METHODS AND RESULTS: Senescence-associated beta-galactosidase (beta-gal) activity was examined in the coronary arteries and the internal mammary arteries retrieved from autopsied individuals who had ischemic heart diseases. Strong beta-gal staining was observed in atherosclerotic lesions of the coronary arteries but not in the internal mammary arteries. An immunohistochemical analysis using anti factor VIII antibody demonstrated that beta-gal stained cells are vascular endothelial cells. To determine whether endothelial cell senescence causes endothelial dysfunction, we induced senescence in human aortic endothelial cells (HAECs) by inhibiting telomere function and examined the expression of intercellular adhesion molecule (ICAM)-1 and endothelial nitric oxide synthase (NOS) activity. Senescent HAECs exhibited increased ICAM-1 expression and decreased eNOS activity, both of which are alterations implicated in atherogenesis. In contrast, introduction of telomerase catalytic component significantly extended the life span and inhibited the functional alterations associated with senescence in HAECs. CONCLUSIONS: Vascular endothelial cells with senescence-associated phenotypes are present in human atherosclerotic lesions, and endothelial cell senescence induced by telomere shortening may contribute to atherogenesis. PMID- 12564114 TI - [Induction of left ventricular remodeling and dysfunction in the recipient heart following donor heart myocardial infarction: new insights into the pathological role of tumor necrosis factor-alpha from a novel heterotopic transplant-coronary ligation model]. AB - BACKGROUND: Neurohormonal and cytokine activation after acute myocardial infarction contribute to cardiac remodeling. This study aimed to examine the effects of tumor necrotic factor (TNF)-alpha and angiotensin II on cardiac remodeling and dysfunction after acute myocardial infarction. METHODS AND RESULTS: We performed isogenic heterotopic cardiac transplantation and simultaneous coronary ligation to produce myocardial infarction in the donor heart, and to evaluate the hearts of both donors and recipients in Lewis rats. The recipients in the ligation group showed significant body-weight loss, hyperthermia, tachycardia, hypotension and leukocytosis at day 7. A significant decrease in left ventricular fractional shortening and + dP/dt, and a significant increase in left ventricular enddiastolic dimension/body weight and left ventricular enddiastolic pressure were also observed in the recipient hearts in the ligation group at day 7. With the exception of the increased perivascular fibrosis, these recipient responses were no longer seen at day 21. TNF-alpha was significantly elevated not only in the plasma but also in the recipient hearts in the ligation group at day 7. In contrast, angiotensin II was significantly increased only in the infarct region of the donor hearts, but not in the plasma. Further, the recipients' transient left ventricular remodeling and dysfunction were completely abolished by the intravenous administration of chimeric TNF-alpha soluble receptor. CONCLUSIONS: We developed a novel heterotopic cardiac transplantation-coronary ligation model capable of inducing myocardial infarction in the absence of downstream hemodynamic effects, and allowing differential quantification of indexes of cardiac remodeling in vivo, such as the local and remote effects of angiotensin II and TNF-alpha on cardiac remodeling. Modification of activated cytokines, such as TNF-alpha induced by cardiac ischemic stress, might be a beneficial strategy for the treatment of cardiac dysfunction and subsequent cardiac remodeling after acute myocardial infarction. PMID- 12564115 TI - [Opening of ATP-sensitive potassium channel attenuates cardiac remodeling induced by chronic inhibition of nitric oxide synthesis]. AB - OBJECTIVES: We examined whether the ATP-sensitive potassium(KATP) channel openers (KCOs) block myocardial hypertrophy, and whether the 70 kDa S6 kinase(p70S6K)-or extracellular signal-regulated kinase(ERK)-dependent pathway is involved. BACKGROUND: Chronic inhibition of nitric oxide (NO) synthesis induces cardiac hypertrophy independent of blood pressure by increasing protein synthesis in vivo. KCOs attenuate calcium overload and confer cardioprotection against ischemic stress, and thereby prevent myocardial remodeling. METHODS: Twelve Wistar-Kyoto rat groups underwent 8 weeks of the drug treatment in combination with NO synthase inhibitor N omega-nitro-L-arginine methyl ester (L-NAME), an inactive isomer D omega-nitro-L-arginine methyl ester(D-NAME), KCOs(nicorandil; 3 and 10 mg/kg/day, or JTV-506; 0.3 mg/kg/day), or a KATP channel blocker glibenclamide. L-NAME was also treated with hydralazine, p70S6K inhibitor (rapamycin) or MAP kinase kinase inhibitor(PD98059). Finally, left ventricular weight-to-body weight ratio(LVW/BW) was quantified followed by histological examinations and kinase assay. RESULTS: L-NAME increased blood pressure and LVW/BW compared with the control. KCOs and hydralazine equally cancelled the increase in blood pressure, whereas only KCOs blocked the increase in LVW/BW and myocardial hypertrophy induced by L-NAME. The L-NAME group showed both p70S6K and ERK activation in the myocardium compared with the control(2.3-fold and 2.0-fold, respectively), which was not reversed by hydralazine. Selective inhibition of either P70S6K or ERK blocked myocardial hypertrophy. KCOs prevented the increase in activity only of p70S6K. Glibenclamide reversed the effect of nicorandil in the presence of L-NAME. CONCLUSIONS: KCOs modulate p70S6K, not ERK, to attenuate myocardial hypertrophy induced by chronic inhibition of nitric oxide synthesis in vivo. PMID- 12564116 TI - [Combination of oral endothelin-A receptor antagonist and oral prostacyclin analogue is superior to each drug alone in ameliorating pulmonary hypertension in rats]. AB - OBJECTIVES: To investigate whether the combination of an oral endothelin (ET)-A receptor antagonist and an oral prostacyclin(PGI2) analogue is superior to the single use of each drug alone for treating pulmonary hypertension(PH). BACKGROUND: Treatment with intravenous PGI2 or an ET-A receptor antagonist was effective for PH; however, the effect of both administrations is unclear. METHODS: We administered the oral ET-A receptor antagonist TA-0201 and/or the oral PGI2 analogue beraprost sodium(BPS) to monocrotaline-induced PH rats for 19 days in the following groups: normal rats with vehicle treatment (Control group), PH rats with vehicle treatment (PH group), PH rats with TA-0201 treatment (PH + TA group), PH rats with BPS treatment (PH + BPS group), and PH rats with TA-0201 and BPS treatment (PH + TA + BPS group). RESULTS: Right ventricular systolic pressure and Pp/Ps were markedly higher in the PH group than in the Control group. The increased right ventricular systolic pressure and Pp/Ps were significantly and comparably depressed in the PH + TA group and PH + BPS group; it was more greatly depressed in the PH + TA + BPS group than in the groups with each drug alone. The indices of right ventricular hypertrophy showed the same tendency as the increase in right ventricular systolic pressure in the five groups. The expression of beta-myosin heavy chain mRNA in right ventricle was markedly augmented in the PH group; the enhancement was inhibited in the PH + TA + BPS group to the greatest degree. Medial wall thickness of the pulmonary artery was markedly increased in the PH group; the increase was depressed in PH + TA + BPS group. Combined treatment also ameliorated PH even if it started from the post-onset of PH. CONCLUSIONS: The combination of an oral ET-A receptor antagonist and an oral PGI2 analogue is superior to the single use of each drug alone in inhibiting the progression of PH. PMID- 12564117 TI - [A 55-year-old man showing sudden disappearance of Q wave]. PMID- 12564118 TI - Investigation of time-dependency of intracranial brain shift and its relation to the extent of tumor removal using intra-operative MRI. AB - The object of the paper is to investigate intra-operative brainshift and its relation to the extent of tumor removal. Repeated T1w 3D datasets were acquired at different time points intra-operatively (T0; T1; T2...Tx) using a vertical open 0.5T MR scanner in six patients with intracranial tumor. An offline analysis with initial linear registration, intensity adjustment and finally nonlinear registration of the first versus subsequent time points (T0/T1; T0/T2...To/Tx) was performed, yielding a 3D displacement vector field that describes the brainshift. Brainshift was analysed qualitatively and quantitatively. A semi automatic segmentation technique was used for calculation of the tumor size and the size of tumor remnants. Semi-automatic segmentation was reliable in all but two cases. Segmentation was difficult and unreliable in astrocytomas grade II. The shift basically followed gravity. The major shift reached levels up to 25 mm. Significant shift was observed at the first time point (T0). Intra-operative brainshift can be analysed qualitatively and also captured quantitatively. Neuronavigation that is based on pre-operatively acquired datasets is associated with a significant risk of surgical morbidity at a very early time point. Parallelisation on a workstation cluster may reduce computation time so that information about the displacement can facilitate updated navigation. PMID- 12564119 TI - Dual blockade of mitogen-activated protein kinases ERK-1 (p42) and ERK-2 (p44) and cyclic AMP response element binding protein (CREB) by neomycin inhibits glioma cell proliferation. AB - Several growth factors and their receptors are expressed in inappropriately high abundance in gliomas and are further upregulated during the transition from low- to high-grade malignancy. In glioma cells growth factors induce expression of mitogen-activated protein kinase (MAPK) pathways. Here we report that neomycin restrained glioma cell proliferation in vitro by inhibition of p42/44 MAPK and the cyclic AMP element binding protein (CREB)-directed transcription pathways. Since alteration of gene transcription by inhibition of specific transcriptional regulatory proteins has important therapeutic potential, neomycin offers great promise for treating cancer and other diseases associated with a sustained MAPK activity. PMID- 12564120 TI - Posterior microendoscopic surgical approach for the degenerative cervical spine. AB - We describe the advantages and clarify the technical key points of a microendoscopic, minimally invasive technique to the posterior surgical approach for cervical degenerative disease. The authors studied the microendoscopic posterior approach using the METRx system in both cadaver models and in clinical cases. This new technique needs only a small surgical route thus reducing damage to the paraspinous muscles. Moreover, this technique provides a clear view of the operating points, because of the oblique view angle of the endoscope. This technique is feasible for not only radiculopathy but also myelopathy caused by segmental canal stenosis. Posterior cervical decompression with this system is technically feasible and should be beneficial for reducing post-operative morbidity and spine deformity. This report deals with cases of cervical radiculopathy and segmental canal stenosis operated on with this system as well as the key points of this surgical procedure. PMID- 12564121 TI - Linezolid: implications for neurosurgical infections. AB - Nosocomial infections affect a significant number of intensive care unit (ICU) patients including those in the neurosurgical ICU. Gram-positive organisms are responsible for many of these infections and often these pathogens are resistant to some of the older antimicrobial agents. Two new classes of antibiotics have been developed: streptogramins and oxazolidinones. Linezolid is an oxazolidinone, which has been shown to be effective against methicillin- and vancomycin resistant Gram-positive pathogens. It may be administered orally or parenterally, and displays favorable pharmacokinetic properties, with rapid and complete absorption after oral administration. Linezolid is generally well tolerated with mild gastrointestinal related adverse effects. Linezolid provides a useful alternative in the treatment of Gram-positive infections, particularly those caused by resistant organisms. It has tremendous clinical utility, especially in the ICU where infections and multi-drug resistant rates are high and treatment options become limited. PMID- 12564122 TI - Nocardial cerebral abscess: report of three cases and review of the current neurosurgical management. AB - Nocardia asteroides cerebral abscesses are rare but challenging intracranial lesions. Early diagnosis, institution of appropriate antimicrobial therapy, lack of underlying systemic disease and limited intracranial disease are recognized factors leading to good outcome. However, the optimal treatment approach has not been established and nocardial brain abscesses have been managed either conservatively, with steroetactic aspirations or with open craniotomy and enucleation. We present three cases of Nocardia asteroides cerebral abscesses cured only after neurosurgical enucleation, and discuss the current management alternatives and conclude that a more aggressive approach than that currently preferred for this entity may be more effective. PMID- 12564123 TI - Nitric oxide levels in rat cortex, hippocampus, cerebellum, and brainstem after impact acceleration head injury. AB - Nitric oxide (NO) is a potential mediator of secondary brain injury in the settings of cerebral ischemia and inflammation. Traumatic brain injury (TBI) alters the levels of stable end products of NO metabolism. We investigated these changes and attempted to identify brain regions that were unique with regard to NO production in the period immediately after TBI. The experiment involved assaying nitrite-nitrate concentrations in the rat cortex, cerebellum, hippocampus, and brainstem after impact-acceleration head injury. Five rats comprised the sham-operated (control) group, five sustained mild head injury (MHI), and five sustained severe head injury (SHI). There was a uniform decline in the tissue concentrations of NO metabolites in all four brain regions in both injured groups. There were no significant differences in the concentrations of NO metabolites among the various sites tested in the MHI group; however, there appeared to be a relationship between degree of decline in NO levels and amount of trauma sustained by a given region in the SHI group. In these rats, NO dropped to the lowest levels in the brain region where the direct trauma was most severe. The results suggest that nitrite-nitrate levels in these four brain regions fall below normal in the first 5 min after impact trauma. This decrease may, in part, be related to reduced activity of all nitric oxide synthase isoforms, which would cause a drop in the levels of NO metabolites. We believe that this decline may be linked to, and may even cause, the global decrease in cerebral blood flow that occurs in the initial stages of TBI. PMID- 12564124 TI - The morphological and neurochemical effects of diffuse brain injury on rat central noradrenergic system. AB - The central noradrenergic system is widely distributed throughout the brain and is closely related to spontaneous motility and level of consciousness. The study presented here evaluated the morphological as well as neurochemical effects of diffuse brain injury on the central noradrenergic system in rat. Adult male Sprague-Dawley rats were subjected to impact-acceleration brain injury produced with a weight-drop device. Morphological changes in locus coeruleus (LC) neurons were examined by using immunohistochemistry for dopamine-beta-hydroxylase, and norepinephrine (NE) turnover in the cerebral cortex was measured by high performance liquid chromatography with electrochemical detection. The size of LC neurons increased by 11% 24 h after injury but had decreased by 27% seven days after injury. Axons of noradrenergic neurons were swollen 24 h and 48 h after injury but the swelling had dwindled in seven days. NE turnover was significantly reduced seven days after injury and remained at a low level until eight weeks after injury. These results suggest that focal impairment of axonal transport due to diffuse brain injury causes cellular changes in LC and that the neurochemical effect of injury on the central noradrenargic system lasts over an extended period of time. Chronic suppression of NE turnover may explain the sustained behavioral and psychological abnormalities observed in a clinical situation. PMID- 12564125 TI - Effects of elevated ICP on brain function: can the multiparametric monitoring system detect the 'Cushing Response'? AB - The 'Cushing Response' is a significant phenomenon associated with elevated ICP. The purpose of our study was to examine the effects of the intracranial hypertension level and duration on the cerebral tissue physiology, using a Multiprobe assembly (MPA). The parameters monitored simultaneously included ICP, CBF, mitochondrial NADH redox state, extracellular K+ and H+ levels, DC potential and ECoG, calculated CPP and blood pressure. Two groups of rats were used. In one group, ICP was elevated to 50-60 mmHg for 13-15 min and, in the second group, ICP was elevated to 20 mmHg for 30 min. The results show that ICP of 50-60 mmHg led to CPP reduction below the lower limits of autoregulation. However, ICP of 20 mmHg, even for a prolonged period of time is completely tolerated. Additionally, we found that the 'Cushing Response', developed in the moderate treatment (ICP = 20 mmHg) is beneficial, assuring high CBF levels under intracranial hypertension. Furthermore, CBF and CPP monitoring, apparently, are not sufficient for autoregulation assessment; more parameters are needed. PMID- 12564126 TI - Multimodality neuroimaging evaluation improves the detection of subtle cortical dysplasia in seizure patients. AB - The purpose of this study is to investigate if multimodality neuroimaging evaluation increases the detection of subtle focal cortical dysplasia as part of an epilepsy surgery evaluation. Three patients with normal magnetic resonance imaging and histopathological findings of focal cortical dysplasia were reviewed. Their magnetoencephalography recordings were performed on whole-head magnetoencephalography system. Magnetic resonance images were re-evaluated with special inspection in limited regions guided by magnetoencephalography spike localization. Two patients had ictal and interictal single photon emission computed tomography study after administration of Tc99m ECD. In two patients we found tiny focal abnormalities including slightly increased cortical thickness and blurred gray-white matter junction at the locations of interictal events after re-evaluation of the MR images indicating focal cortical dysplasia. The third patient showed focal atrophic change. All patients are seizure free after surgery. Both ictal and interictal single photon emission computed tomography showed hyperperfusion in the dysplastic cortex regions. Multimodality neuroimaging study can improve the detection of focal cortical dysplasia. Normal magnetic resonance images should be re-evaluated for subtle signs of focal cortical dysplasia especially when magnetoencephalography recording demonstrate focal epileptic discharges. PMID- 12564127 TI - Effects of myasthenia gravis patients' sera with different autoantibodies on slow K+ current at mouse motor nerve terminals. AB - The antibodies against pre-synaptic membrane receptor (PsmR) and acetylcholine receptor (AChR) in serum samples of myasthenia gravis (MG) patients and healthy donors were tested by enzyme-linked immunosorbent assays (ELISA). The serum samples of eight MG patients with different autoantibodies and those of six healthy donors without these two kinds of autoantibodies were collected to investigate their effects on the peri-neurially recorded membrane currents at mouse motor nerve terminals. After inhibition of both fast and Ca(2+)-dependent K+ currents by tetra-ethylammonium (TEA), a positive wave was revealed, which was a balance of the slow K+(Ik,s) and Ca2+ currents (ICa). Application of anti-PsmR antibody negative MG sera and healthy donor sera, whether anti-AChR antibody positive or negative, did not affect the positive wave. However, the positive wave shifted to prolonged Ca(2+)-plateau when adding two of four anti-PsmR antibody positive serum samples from MG patients, indicating an inhibition of Ik,s by anti-PsmR antibody positive sera. Meanwhile, all serum samples derived from either patients or healthy donors did not affect INa. PMID- 12564128 TI - Primarily generalized seizures are more effective than partial seizures in arousing patients from sleep. AB - The purpose of this study was to determine the characteristics of generalized and partial seizures which awaken patients from sleep, using a retrospective review of intracranial EEG recordings in 8017 electrographic and 7571 clinical seizures in 172 patients undergoing evaluation for epilepsy brain surgery. Seizure onset during sleep followed by awakening occurred in 99% of 308 seizures in 22 patients during daytime naps. Four events consisted of spontaneous awakening followed by the seizure. In contrast electrographic seizures almost never awakened the patient if they were partial in onset (0.02% temporal, 0% frontal), but did so 26% of the time if they were generalized (p < 0.001). Conversely, generalized clinical seizures awakened the patient only 0.3% of the time (p < 0.001) versus 3% for temporal and 6% for frontal lobe. Partial and generalized seizures differ during sleep. Partial seizures do not awaken until they propagate outside the lobe and evolve into a clinical seizure. Generalized seizures when only electrographic, include wake-regulating structures at their onset (presumably thalamus, hypothalamus, brainstem). Our results suggest that rather than sleep transitions being a facilitatory cause of seizures, seizures awaken us from sleep via endogenous stimulation of the brain's sleep/wake structures. This pathway information may be relevant to planning epilepsy brain surgery. PMID- 12564129 TI - Chronic ischemic stroke model in cynomolgus monkeys: behavioral, neuroimaging and anatomical study. AB - Previous nonhuman primate stroke models have employed temporary occlusion of arteries, had limited behavioral testing and imaging, and focused on the short term outcome. Our goals were 1. to develop a stable model of chronic stroke in the nonhuman primate, 2. to study in vivo the long-term biochemical changes in the area adjacent to the infarct, using proton magnetic resonance spectroscopy (H MRS), and 3. evaluate these changes in relation to the histopathological effects of stroke. Four adult cynomologous monkeys had an occlusion of the M1 segment of the right MCA. Behavioral tests included a clinical rating scale, motor planning task, fine motor task, and activity monitoring. Eight months afterwards, MRI and 1H MRS were performed. Following the imaging studies the monkeys were perfused transcardially, their brains extracted and processed. Nissl staining and immunohistochemistry for neuronal markers (NeuN) were performed and used to measure the lesion volume and neuronal optical density (OD). All animals developed a left hemiparesis and were unable to perform a fine motor task with the left hand. There was a significant (31%) decline in the motor planning ability with the nonparetic extremity. Monkeys displayed a stooped posture, episodes of rotation to the side of the lesion, partial left hemianopsia, and transient changes in activity. The clinical signs improved over the first 6-8 weeks but the deficits remained stable for the remaining six months of follow up. MRI demonstrated a subcortical and cortical infarction in the right MCA distribution. 1H MRS data detected a significant decrease in the N-acetyl aspartate (NAA)/creatine (Cr) ratio in the area adjacent to the infarction (VOl St) compared to a mirror area in the contralateral hemisphere (VOl-Co). Histopathological measurements revealed a significant decline in neuronal cross sectional area and neuronal optical density in the region of the VOl-St. We established a stable and reproducible model of chronic stroke in the MCA distribution, in the macaque monkey. Our data indicate that NAA detected by 1H MRS can be used to measure neuronal loss in vivo and help target this area for intervention. Our model may be particularly suitable for studies testing the effects of therapeutic strategies involving neural or stem cell transplantation, trophic factors or gene therapy. PMID- 12564130 TI - Temporal and spacial changes of highly polysialylated neural cell adhesion molecule immunoreactivity in amygdala kindling development. AB - To investigate the migration of neural stem cells as well as neural plastic changes in epileptic brain, spaciotemporal expression of immunoreactive highly polysialylated neural cell adhesion molecule (PSA-NCAM) was examined in amygdala kindling development of rat. The neural migration and synaptic remodeling detected with PSA-NCAM staining occurred in dentate gyrus of hippocampus, subventricular zone and pyriform cortex with amygdaloid kindling in generalized seizure but not in partial seizure. Although PSA-NCAM positive dendrite in dentate gyrus was minimally found in the control brain, it extended slightly in animals with partial seizure, and greatly toward the molecular layer with generalized seizure. Thus, the migration of neural stem cells as well as neural plastic changes were specially and temporally different between brain regions depending on different kindling stages. These changes may mainly contribute to the reorganization of neural network in epileptic brain. PMID- 12564131 TI - Ischemia-induced changes in monocarboxylate transporter 1 reactive cells in rat hippocampus. AB - This study aims to demonstrate the responses of monocarboxylate transporter 1 (MCT1) immunoreactive cells to transient global ischemia in rat hippocampus using confocal and electron microscopy. The MCT1 staining in CA1 pyramidal cells of the sham-operated controls appeared evenly distributed. Most of the MCT1 immunoreactive products were associated with the cell surface; however, some intracellular reaction products are also found. This pattern of stain was not altered in the first three days after an ischemic episode. As the neuronal demise progressed, the MCT1 immunoreactive cells became patchy in the 21-day post ischemic rats. Besides the neuronal labeling, MCT1 immunoreactivity was found in astroglia, in endothelial cells and in the adjacent ependymal lining. The latter exhibited intense labeling both in the acute and long-term surviving rats. These data suggest that MCT1 plays a role in the initial and long-term neuronal survival in the hippocampus. PMID- 12564132 TI - Magnetically evoked EMGs in rats. AB - Magnetic stimulation of the brain and spinal cord was carried out in rats to record electromyogram (EMGs) from the gastrocnemius. A figure-eight coil was set over the middle of the dorsum, and shifted from the cervical vertebrae to the sacrum. The motor evoked potentials (MEPs) with 4.8 msec latency by transcranial magnetic stimulation and the descending wave with 4.7 msec latency by C3-C4 stimulation were recorded. In evoked EMGs by magnetic stimulation over T9-T10, L4 L5, S2-S3 and Ca2-Ca3 spinal cord levels, the causes of these two evoked components with short (1.5 msec) and long (4.1 msec) latencies were estimated to be the eddy current generated from the rostral to the caudal portion of the spinal cord. With the increase in magnetic stimuli, the relative sizes and disappearance of H- and M-like responses were comparable with the ordinary M- and H-responses in electrically evoked EMGs. The magnetic stimulation of the spinal cord activated the sciatic nerve at their vertebral exit, because the latencies of the H- and M-responses were constant despite the changing stimulus sites. Although magnetic stimulation with the figure-eight coil can be focused on the target, it is necessary to take into consideration the influence of the eddy current flowing in the body. PMID- 12564133 TI - Multi-organ cavernous hemangiomas: case report. AB - Hemangiomas are rare benign tumors of vascular origin and multiple location of these tumors in different organs is extremely unusual. We report a case with multiple hemangiomas, characterized by calvarial, hepatic and suspicious costal involvement. Organ involvement and multiplicity of the neoplasm made our case very interesting and distinct. Our search of the English literature has revealed that no similar case has ever been published. PMID- 12564134 TI - Optimal dosing as a necessary condition for the efficacy of hyperbaric oxygen therapy in acute ischemic stroke: a critical review. AB - The effectiveness of hyperbaric oxygen therapy (HBOT) in clinical and experimental acute ischemic stroke (AIS) has been controversial for many years. However, in the literature, no data was found on the dose/effect of HBOT in patients with AIS. We analyzed retrospectively the published data of clinical studies performed in different hyperbaric centers (a total of 265 patients). The dose of HBOT (DHBOT) was calculated considering the product intrabarochamber pO2 (ATA), the duration of a single HBOT exposure (hours), and the number of HBOT treatments. Efficacy of HBOT (EfHBOT) data regarding the number of patients who showed significant clinical improvement of their neurologic status in the course of the treatment HBOT (the percentage of the total number of patients). The level of EfHBOT in each study was compared with a corresponding value of DHBOT. A comparison of the data shows a pronounced tendency for higher values of EfHBOT as the level of the average values of the total DHBOT increases. The coefficient of correlation between these parameters appears to be fairly high (r = 0.92). The maximum possible value of EfHBOT is 100%, which corresponded to the average values of DHBOT at a level of no less than 30 agreed units. The examined data suggest that applying optimal total DHBOT may provide a maximum possible EfHBOT in treating patients with AIS. PMID- 12564135 TI - Evaluating brain tolerability to carotid artery occlusion. AB - An objective and practicable method to evaluate the brain tolerance to unilateral carotid artery occlusion was attempted. Ten consecutive cases were studied. Endovascular trial balloon occlusion of the unilateral carotid artery in conjunction with single-photon emission computed tomography (SPECT) as well as digital subtraction angiography (DSA) and clinical neurological evaluation was routinely performed for those patients who might undergo permanent occlusion of the internal (ICA) and/or common carotid artery (CCA). The result of SPECT was analyzed semi-quantitatively and compared with the baseline data as well as the data from contralateral side. The relative symmetry index (rS) of side-to-side radioactivity counts relevant to the baseline was calculated. One subject failed the balloon occlusion test (BOT), even though an angiographically adequate collateral circulation was observed. The rS of the patient was 74.5%. The remaining patients passed the 45-min BOT without any neurological deficiency induced. Their value of rS was 97.8% +/- 4.4%. Based the BOT results, the left ICA, CCA and external carotid artery were excised in one patient, CCA-ICA reconstructions after carotid occlusions were performed in three. The carotid arteries were saved in five. For one patient, the ICA was occluded spontaneously during the BOT. There were no neurological impairments developed after the surgery. With this BOT technique, clinically silent areas of decreased perfusion might be detected. We suggest it be a routine preparatory to carotid manipulations. PMID- 12564136 TI - Cytotoxicity of ventricular cerebrospinal fluid from Parkinson patients: correlation with clinical profiles and neurochemistry. AB - Other investigators have reported that the cerebrospinal fluid (CSF) from patients with Parkinson's disease (PD) might contain endogenous dystrophic factors. Using CSF samples drawn from individual PD patients during surgery, we investigated the toxic effect of ventricular CSF (vCSF) on the growth of PC12 cells and the correlation between the clinical profiles of the patients and CSF neurochemistry. Ventricular CSF samples from 28 patients with PD or essential tremor (ET) were collected during ventriculography for stereotactic pallidotomy or thalamotomy. PC12 cells were incubated with 20% vCSF from both clinical groups for up to 72 h. Microdialysis was used to analyze four neurochemical parameters (glucose, lactate, pyruvate, and glutamate) in each vCSF sample. We observed that vCSF drawn from PD patients exerted nonspecific growth inhibition on PC12 cells in a time-dependent manner. The growth inhibitory action of PD-vCSF decreased significantly after heat treatment. Microdialysis demonstrated no statistical differences between PD and ET samples among the four parameters studied. In addition, PC12 cell survival after 72 h incubation with PD-vCSF correlated with no neurochemical parameter or individual clinical profile (age, onset age, duration of disease, Hoehn & Yahr stage, disease progression rate), except for a slight correlation between vCSF and disease progression rate in heat treated samples from female patients. One or more endogenous cytotoxic factors in PD-vCSF inhibit PC12 cell growth. This factor or factors are partially sensitive to heat which suggests proteins or peptides as possible agents. The cytotoxic effect of PD-vCSF did not directly correlate with any clinical profiles studied or energy metabolism of PD brain. PMID- 12564138 TI - Ready as ready can be. Many healthcare IT departments are already prepared for HIPAA. PMID- 12564137 TI - 11th-hour HIPAA: how can you meet the deadlines? PMID- 12564139 TI - Infrastructural considerations for supporting POC devices. Healthcare executives must focus on creating integration-friendly environments for the most effective use of mobile devices. AB - Creating an integration-friendly infrastructure is the best way to prepare for current as well as next-generation POC devices. Fortunately, hassle-free system integration is needed throughout healthcare today for reasons beyond the domain of POC devices, so integration progress already made in other areas helps pave the way for POC devices. Healthcare IT professionals, as much or more than any other IT group, have had to deal with the challenges of integrating disparate systems. When considering deployment of POC devices, you will want to make sure they adhere to open industry standards. In this way, these important devices won't add to the problem of disparate systems, but will contribute to the solution. We see XML and Web services as being especially important to the future of healthcare delivery and administration. XML and Web services, along with powerful orchestrators, can provide an ever-richer collection of clinical data to be delivered to, and received from, the POC devices that will become an ever more important addition to the physician armamentarium. PMID- 12564140 TI - Push technology in the pharmacy. Clinical decision support helps the pharmacy department of a South Carolina medical center automate monitoring of medication effectiveness. PMID- 12564141 TI - BPM: antidote to inefficiency. Business process management serves as a foundation for effective workflow automation in healthcare. PMID- 12564142 TI - Remote access for physicians. SSL VPNs offer advantages for healthcare organizations that want to provide mobile physicians with secure access. PMID- 12564143 TI - Rx for password headaches. Biometric authentication solution lets physicians be their passwords. PMID- 12564144 TI - Productivity gains of speech-recognition technology. Organizations that gain the most will be those armed with the facts, tempered by reasonable expectations. PMID- 12564145 TI - Voice recognition goes home. Arizona physical therapy practice successfully uses voice recognition with the advantage of portability. PMID- 12564146 TI - What works. An inside look at outsourcing. Southern California healthcare system finds financial and performance pluses in IT outsourcing. PMID- 12564147 TI - What works. Go with the flow. Medical group overcomes geographic obstacles to improve workflow. PMID- 12564148 TI - Instructive resistance. PMID- 12564149 TI - Crime scene conflict. Homicide, strong-arm robbery. PMID- 12564150 TI - United we respond. PMID- 12564151 TI - The lifesaver project. PMID- 12564152 TI - Professionalism. PMID- 12564153 TI - Backseat denial. PMID- 12564154 TI - The value of ethical behavior. PMID- 12564155 TI - Putting the customer first. PMID- 12564156 TI - Patient ventilation and oxygenation. A new day is dawning. AB - Maintaining an airway and providing adequate ventilation and oxygenation to the patient can be challenging in the prehospital environment. Ventilation and oxygenation are a complex series of interactions between the patient, EMS providers and emergency airway equipment. Routine ventilation techniques carry significant risk of long-term complications. New ventilatory equipment is available to perform this function and provide verification of its effectiveness. This opportunity for improved patient care is available to all EMS providers and sets new standards for delivery of ventilation and oxygenation. The technical methods available to EMS personnel vary considerably and are reviewed in this article. PMID- 12564157 TI - The difficult airway. PMID- 12564158 TI - Searching for the evidence behind EMS. AB - This is an exciting time to be involved in EMS. EMTs and paramedics can derive a great deal of satisfaction by assuring that they have offered their patients the best care available, based not only on their personal expertise, but also on ample external scientific evidence. EBM is a sound and fairly simple way to merge the two. Take your prehospital practice to the next level--give evidence-based medicine a try. PMID- 12564159 TI - Caring for victims of domestic violence and sexual assault. PMID- 12564160 TI - 360-degree quality management. PMID- 12564161 TI - EMS in Ghana. PMID- 12564162 TI - Mean age of mother, 1970-2000. AB - OBJECTIVES: This report presents trends in the mean age of mothers giving birth for the United States for the last three decades. Data are presented by live birth order, race, Hispanic origin, and State of residence of the mother. METHODS: Descriptive tabulations and graphs of the trends in the mean age of mother are discussed. RESULTS: The mean age of mother has increased steadily in the United States over the last three decades. Mothers having their first and second live births had the largest increase in mean age. Among racial and Hispanic origin subgroups, Japanese women had the highest mean age in 2000. Puerto Rican women had the lowest mean age. Massachusetts had the highest mean age in 2000 and Mississippi had the lowest. PMID- 12564163 TI - [Retinitis pigmentosa: pathogenic progress and therapeutic hopes]. PMID- 12564164 TI - [Neglected aspects of the antialcoholic iconography: lantern slides and film strips]. PMID- 12564165 TI - [Role of Fallopian tube in fertilization]. AB - Fallopian tubes are complex organ where fertilization occurs after inverse migration of spermatozoa and oocyte. However, tube cannot be restricted to be fertilization site. The different tubal segments represent specific microenvironments to favour capacitation and fertilization process and to support early embryonic development. Tubal physiology permits tubal transport of gametes and embryon on adequate time by variations of smooth muscles contractility among tubal segments and during menstrual cycle. Tubes should control the number of competent spermatozoa present on fertilization site. Composition and viscosity of tubal fluid also vary along the tube and during cycle. Specific components of tubal fluid such as HuOGP interfere with gamete maturation, fertilization process and early embryonic development. Alteration of part of these functions could explain some tubal infertilities leading with normal tube morphology. PMID- 12564166 TI - [Acute salpingitis]. AB - The incidence of acute salpingitis which are only one aspect of pelvic inflammatory disease has decreased during the last 20 years and more and more patients have mild symptoms. Consequences of this evolution are uncertainty for the diagnosis without laparoscopic proof of the pelvic inflammatory disease, ambulatory treatment with inappropriate regimen increasing the risk of chronical disease and long term sequelae. So laparoscopy must remain a standard in diagnosis and treatment especially in young childless women. Follow up of the medical treatment is necessary to assess its effectiveness and its compliance, to treat the sexual partner, and to provide informations about preventive measures to avoid relaps. PMID- 12564167 TI - [Imaging in tubal infertility]. AB - Since the appearance of the new imaging techniques (ultrasonography, CT MRI) the hysterosalpingography has lost many indications. Therefore, the hysterosalpingography remains one of the most useful investigations in the assessment of tubar infertility; provided that it is not restricted to only demonstrate the permeability of Fallopian tubes. The high quality of the pictures and their sharp analysis is one of the major factor of both clinical and therapeutical decisions. The selective uni or bilateral salpingography provides gynecologists with the best investigations for therapy of tubar infertility and for the tubar sterilisation. PMID- 12564168 TI - [Surgery of tubal lesions with favourable prognosis]. AB - Presently, open microsurgery and laparoscopic microsurgery must be used for the treatment of tubal lesions with favourable prognosis (70% of proximal tubal lesions and 45% of distal tubal lesions) because, in these lesions, the results are better than these obtained by in vitro fertilization. Essential principles of microsurgery must be respected by the tubal surgery. PMID- 12564169 TI - [Diagnosis of ectopic pregnancy]. AB - The knowledge of ectopic pregnancies risk factors (intra-uterine device; past history of pelvic inflammatory disease; history of tubal pathology and tubal surgery; positive chlamydiae serology; and smoking), the high sensitivity and specificity of radioimmunological assay of beta-HCG and the availability of high resolution sonography using vaginal probes had improved the routinely diagnosis of ectopic pregnancy. Today, diagnosis was made with beta-HCG and vaginal echography. Uterine vacuity, peritoneal liquid and latero-uterine masse represent the major ultrasound signs. However, diagnosis can be uncertain below discriminative zone of beta-HCG (< 1,500 to 2,000 UI/L). Other new diagnostic markers are also described, as progesterone, creatine kinase, VEGF and CA 125. The place of ponction of the Douglas, endouterine curettage, hysteroscopy and laparoscopy are also defined for diagnosis of ectopic pregnancy. PMID- 12564171 TI - [Tubal sterilization]. AB - A recent French law, passed on July 4th, 2001, authorizes a person of age to voluntarily undergo a sterilisation on her request. The practitioner has the legal obligation to inform the person of the consequences and the possible complications of the operation. Recent technical progress as well as its assessment on a large epidemiological scale has modified the given data. In this perspective, we are presenting the different and current techniques of tubal sterilisation, their efficiency and their immediate and long-term complications. During the consultation preceding the operation, the doctor should be certain that the patient (or the couple) is well aware and he will help her (or them) in finding the right contraception. Finally, we will mention relevant passages of the new law. PMID- 12564170 TI - [Treatment of ectopic pregnancy]. AB - Ectopic pregnancy, a sometimes mortal complication, currently rests one of the more frequently seen problems in women being treated for sterility. The treatment, classically surgical, is now tending towards a medical treatment by methotrexate. Surgical excision of a tubular ectopic pregnancy is done by laparoscopy in preference to laparotomy, which is reserved for an acute haemoperitoneum. The conservation of the fallopian tube depends on multiple factors. Subsequent fertility is dominated by the risk of a recurrence, making call to in vitro fertilization (IVF) in the case of bilateral tubular lesions. Pharmacotherapy by methotrexate is simple to perform, but demands rigourous surveillance. PMID- 12564172 TI - [Family practice and general medicine]. PMID- 12564173 TI - [Polycythemia vera]. PMID- 12564174 TI - [Tuberculosis]. PMID- 12564175 TI - [Acute pericarditis]. PMID- 12564176 TI - [Cerebrovascular attacks]. PMID- 12564177 TI - [Auditive function disturbances. Diagnostic orientation]. PMID- 12564178 TI - John T. Cacioppo. Award for Distinguished Scientific Contributions. PMID- 12564179 TI - Social neuroscience: understanding the pieces fosters understanding the whole and vice versa. AB - Social science and neuroscience perspectives represent two ends of a continuum of levels of organization studied in psychology. Human behavior as a whole unfolds at social levels of organization, whereas much of the research in psychology has focused on cognitive and biological pieces of this whole. Recent evidence underscores the complementary nature of social, cognitive, and biological levels of analysis and how research integrating these levels can foster more comprehensive theories of the mechanisms underlying complex behavior and the mind. This research underscores the unity of psychology and the importance of retaining multilevel integrative research that spans molar and molecular levels of analysis. PMID- 12564180 TI - David E. Meyer. Award for Distinguished Scientific Contributions. PMID- 12564182 TI - Robert Rosenthal. Award for Distinguished Scientific Applications of Psychology. PMID- 12564181 TI - William T. Newsome. Award for distinguished scientific contributions. PMID- 12564183 TI - Covert communication in classrooms, clinics, courtrooms, and cubicles. AB - Interpersonal expectancy effects refer to the phenomena whereby one person's expectation for another person's behavior comes to serve as a self-fulfilling prophecy. The author examines the history and diversity of this area of inquiry, showing that the expectations of psychological researchers, classroom teachers, judges in the courtroom, business executives, and health care providers can unintentionally affect the responses of their research participants, pupils, jurors, employees, and patients. Using meta-analytic procedures, the author examines (a) moderator variables associated with the magnitude of interpersonal expectancy effects and (b) mediator variables implicated in the communication of interpersonal expectations. The author considers the social importance of the magnitudes of the obtained effects and points out research still needed to clarify issues in the mediation of these effects. PMID- 12564184 TI - Deanna M. Barch. Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 12564185 TI - Donald R. Lynam. Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 12564186 TI - Nicki R. Crick. Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 12564187 TI - Alexander J. Rothman. Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 12564188 TI - Marvin M. Chun. Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 12564189 TI - Julie A. Fiez. Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 12564190 TI - Lisa M. Savage. Award for Distinguished Scientific Early Career Contributions to Psychology. PMID- 12564191 TI - Micheal E. Shafer. Psi Chi/APA Edwin B. Newman Graduate Research Award. PMID- 12564192 TI - Phyllis A. Katz. Award for Distinguished Senior Career Contributions to the Public Interest. PMID- 12564193 TI - Melba J. T. Vasquez. Award for Distinguished Senior Career Contributions to the Public Interest. PMID- 12564194 TI - Complexities of the Latina experience: a tribute to Martha Bernal. AB - This address discusses educational attainment rates among Latinas and factors that militate against their educational and professional achievement. The roles of family, community, discrimination, and strategies to counteract discrimination are discussed. It also contains a tribute to a Latina pioneer in psychology, Martha Bernal. PMID- 12564195 TI - Brian Smedley. Award for Distinguished Early Career Contributions to Psychology in the Public Interest. PMID- 12564196 TI - Jeanne Brooks-Gunn. Award for Distinguished Contributions to Research in Public Policy. PMID- 12564197 TI - Barbara M. Byrne Award for Distinguished Contributions to Education and Training in Psychology. PMID- 12564198 TI - Validating the measurement and structure of self-concept: snapshots of past, present, and future research. AB - Although the study of self-concept has been a topic of great interest and much study in the social sciences for many decades, it is really only in the past 30 years that any fruitful expansion in knowledge of both its theoretical structure and its related measurement has been forthcoming. From three perspectives--past, present, and future--the author presents abbreviated and selected highlights of important construct validity findings related to this research and postulates possible trends and areas of self-concept research yet to be explored. PMID- 12564199 TI - Jessica Henderson Daniel. Award for Distinguished Contributions to Education and Training in Psychology. PMID- 12564200 TI - Virginia Andreoli Mathie. Award for Distinguished Contributions of Applications of Psychology to Education and Training. PMID- 12564201 TI - Building academic partnerships in psychology: the Psychology Partnerships Project. AB - The increased number and mobility of psychology students, the need for a more coordinated curriculum and a more diverse student population in psychology programs, and the need to prepare students for a changing marketplace pose serious challenges for psychology educators. Partnerships among psychology teachers in high schools, community colleges, 4-year colleges and universities, and graduate programs in research universities can help teachers address these challenges effectively. This article outlines how academic partnerships across educational levels can help psychology teachers address educational challenges, examines factors that facilitate the formation and maintenance of these partnerships, and presents the American Psychological Association's successful Psychology Partnerships Project: Academic Partnerships to Meet the Teaching and Learning Needs of the 21st Century, describing the organizational features that enhanced its success. PMID- 12564202 TI - John D. Krumboltz. Award for distinguished professional contributions to knowledge. PMID- 12564203 TI - Encouraging research: make it collegial, enjoyable, and relevant. AB - Researchers are too often trained under circumstances that discourage them from continuing research activities after they graduate. Three hypotheses are suggested that, if confirmed, would lead to a greater number of competent and enthusiastic researchers. (a) Arrange compatible peer groups. Allowing graduate students to form research teams to conduct dissertation studies would provide more social support than the individualistic environment traditionally required does. (b) Make the experience satisfying and enjoyable. Infuse a spirit of playfulness and puzzlement. PMID- 12564204 TI - Frank J. Sullivan. Award for distinguished professional contributions to public service. PMID- 12564205 TI - William R. Safarjan. Award for distinguished contributions to applied psychology as a professional practice. PMID- 12564206 TI - A primer for advancing psychology in the public-sector. AB - Psychologists working in the public sector should, in addition to providing direct services, advocate for systems change. Although many consumers treated in the public sector face a constellation of severe life problems, working to improve the system of care is more difficult than providing treatment. Improving the quality of life of consumers of publicly funded mental health services requires that psychologists become advocates. Four prerequisites to systems change, plus coalition building, legislative advocacy, the work of state psychological associations, and forming alliances between psychologists and nonpsychological community organizations such as Rotary International, are described. In conclusion, 12 orienting ideas are listed for psychologists who want to advocate for social, institutional, and political change. PMID- 12564207 TI - Anne S. Labowitz APA/APAGS distinguished graduate student award in professional psychology. PMID- 12564208 TI - Stanley C. Krippner. Award for distinguished contributions to the international advancement of psychology. PMID- 12564209 TI - Conflicting perspectives on shamans and shamanism: points and counterpoints. AB - Shamans' communities grant them privileged status to attend to those groups' psychological and spiritual needs. Shamans claim to modify their attentional states and engage in activities that enable them to access information not ordinarily attainable by members of the social group that has granted them shamanic status. Western perspectives on shamanism have changed and clashed over the centuries; this address presents points and counterpoints regarding what might be termed the demonic model, the charlatan model, the schizophrenia model, the soul flight model, the degenerative and crude technology model, and the deconstructionist model. Western interpretations of shamanism often reveal more about the observer than they do about the observed; in addressing this challenge, the study of shamanism could make contributions to cognitive neuroscience, social psychology, psychological therapy, and ecological psychology. PMID- 12564210 TI - Elizabeth Lira. APA international humanitarian award. PMID- 12564211 TI - Meredith P. Crawford (1910-2002). PMID- 12564212 TI - The impossibility of parental licensure. PMID- 12564213 TI - Can people with disabilities be parents? PMID- 12564215 TI - Valid inferences from invalid tests? PMID- 12564216 TI - Practicing psychological assessment. PMID- 12564217 TI - [Characteristics of human adaptation in the highlands of Central Asia and the Antarctic]. PMID- 12564218 TI - [Rearrangements of the algorithms of interaction between wave components of EEG in subjects with different mechanisms of brain self-regulation during adaptation to high altitudes]. PMID- 12564219 TI - [Relationships between cerebral blood flow dynamics and bioelectric activity of the human brain in experimental acute hypoxia]. PMID- 12564220 TI - [Assessment of the significance of psychophysiological components in maintaining the activity of operators in high mountain areas]. PMID- 12564221 TI - [Membrane and intracellular components in adaptation to physical factors of mountain regions]. PMID- 12564222 TI - [Effect of short-term intermittent normobaric hypoxia on the regulation of external respiration in humans]. PMID- 12564223 TI - [The role of right- and left-hemispheric structures in speech and memory formation in children]. PMID- 12564224 TI - [Leading characteristics of the maturity of brain bioelectric activity in children 1-7 years of age]. PMID- 12564225 TI - [A model of physiological monitoring in the complex examination of the adaptability of students during their educational activity. Communication 2. The hormonal psycho-autonomic status of adolescents living under different socioecological conditions]. PMID- 12564226 TI - [The problem of cooperation of hand reaction. Communication I. Cooperation of hand reaction to sound and light stimuli in healthy subjects]. PMID- 12564227 TI - [Comparison of leg muscle responses to transcranial magnetic stimulation during standing on stable and unstable supports]. PMID- 12564228 TI - [Changes in the parameters of cellular immunity in patients with right- and left hemispheric ischemic stroke after transcranial electromagnetic stimulations]. PMID- 12564229 TI - [Dynamics of the variables of neonates' physical development and geomagnetic variables over a many-year period]. PMID- 12564230 TI - [The role of voluntary motivation in the tolerability of additional respiratory resistance]. PMID- 12564231 TI - [Cardiorespiratory synchronism in humans]. PMID- 12564232 TI - [Structural and functional features of left heart chambers and hemodynamic response to physical exercise in young athletes]. PMID- 12564233 TI - [Changes in electrocardiographic parameters in males of the European North as markers of climate- and age-related effects]. PMID- 12564234 TI - [Glucose metabolism in healthy subjects and in patients with carbohydrate and lipid metabolism disorders]. PMID- 12564235 TI - [Physiological mechanisms of the effect of modulated kinesotherapy on the functional state of blood circulation]. PMID- 12564236 TI - [Effects of electric stimulations or low-frequency laser irradiation on the pH of the nerve]. PMID- 12564237 TI - Heterosexual clusters of HIV infection. PMID- 12564238 TI - The making of an epidemiologist: John Snow before the episode of the Broad Street pump. PMID- 12564239 TI - Doncaster: the public health response to a local cluster of heterosexually acquired HIV infection. AB - In recent years, clusters of sexually transmitted HIV infection have attracted some media prominence in the UK. We describe our response to a local HIV cluster of five cases that acquired a high media profile. Practical difficulties encountered included: ensuring medical confidentiality in an outbreak control team which included many professional groups; handling a large volume of helpline calls; differing views on how specific health promotion should be; and the impact of summer leave on the control team's continuity. Following publicity, two further cases of HIV infection were found, bringing the total cluster to seven. Both later cases named the man at the centre of the cluster as a sexual partner. Of these two cases, one was local, identified as a direct result of the campaign. The other, living 170 miles distant in London, was identified as a result of increased media awareness. Eventually, the press named the man they thought to be the source case, which the outbreak control team had avoided doing. Good inter disciplinary working made a co-ordinated public health response possible, and the 1,600 helpline calls dealt with served to reassure the public. Over 1,000 of the callers were advised to contact the Genito Urinary Medicine (GUM) department of their local hospital for counselling and testing of whom 772 took the advice. However, the longer-term effect on sexual behaviour was minimal as evidence by a subsequent survey. PMID- 12564240 TI - Description of an HIV patient notification exercise in Essex involving a dental healthcare worker. AB - During 2000, one of the Consultants in Communicable Disease Control (CsCDC) for North Essex Health Authority was advised that a healthcare worker (HCW) in dentistry had recently been diagnosed as HIV positive. As a result of the HCW's history and taking account the Guidance on HIV infected healthcare workers of the United Kingdom Expert Advisory Group on AIDS, North and South Essex Health Authorities embarked on the largest patient notification exercise in the United Kingdom, to date. This involved an examination of 36,000 patient records and resulted in the identification of 5,929 exposed patients. Of these 5,929 patients, 3,825 contacted the health authority helpline and 3,096 were counselled and screened for evidence of HIV infection. All tests were negative. This was also the first patient notification exercise to use salivary samples for patient screening. The exercise cost in excess of 300,000 Pounds, ignoring the opportunity costs of staff having to devote time to the exercise. PMID- 12564241 TI - Effect of incubation temperature on the isolation of thermophilic species of Campylobacter from faeces. AB - Although no large-scale clinical study has been performed, it has been reported that incubation at 37 degrees C gives better isolation rates for all common species of Campylobacter than incubation at 42 degrees C, while also improving the recovery of the more unusual species. In this study, 2,570 faecal samples were examined in four laboratories, using a standardised protocol. Isolation rates of Campylobacter spp. were compared after incubation on modified campylobacter blood-free selective agar at 37 degrees C and 42 degrees C. Campylobacter spp. isolates were made from 185 samples (7.2%); 25 were recovered only at 42 degrees C and three only at 37 degrees C (p < 0.001). There were significantly more colonies at 42 degrees C (p = 0.001). Competing flora were heavier at 37 degrees C, but this did not account for the difference in isolation rates or quantity of growth. It is recommended that cultures for Campylobacter spp. be routinely incubated at 42 degrees C. More specific techniques are required to seek for fastidious campylobacters. PMID- 12564242 TI - Comparison of the performance of lactose and mannitol selenite enriched broths, subcultured to DCA and XLD agars, in the isolation of Salmonella spp. from faeces. AB - Selenite-based enrichment broths using either lactose or mannitol as a carbohydrate source are generally used as selective enrichment media for the isolation of Salmonella spp. from human faeces in the UK, but few studies have compared the relative efficacy of the available formulations. A variety of solid media is used for the routine subculture from these selective broths, but similarly we have been unable to find published evidence as to which medium performs best. Four thousand and nineteen faecal samples were examined in four laboratories and the isolation rates of Salmonella spp. from lactose (LS) or mannitol selenite (MS) broths, plated onto either xylose lysine desoxycholate agar (XLD) or desoxycholate citrate agar (DCA) were compared. MS performed significantly better than LS (p = 0.02), recovering 95 salmonellae compared with 87. No significant difference in isolation rates was found between XLD and DCA, although colonial appearances of suspected salmonellae on XLD were much more specific, resulting in significantly fewer colonies having to be selected for supplementary testing (p < 0.001) and so reducing confirmatory work. An opportunity exists to simplify holdings of media by choosing to use the MS/XLD combination. PMID- 12564243 TI - Microbiological quality of take-away cooked rice and chicken sandwiches: effectiveness of food hygiene training of the management. AB - During August 2001 a microbiological study of ready-to-eat cooked rice from take aways and of chicken sandwiches made on the premises from sandwich bars was undertaken. The intention was to identify risk factors in the production, storage and handling of cooked rice and sandwiches, and to establish their effect on microbiological quality. Examination of cooked rice revealed that the majority of samples (87%; 442 of 508) were of satisfactory/acceptable microbiological quality; 50 (10%) were unsatisfactory, and 16 (3%) were of unacceptable quality due to Bacillus cereus and/or other Bacillus spp in excess of 10(5) cfu/g. The microbiological quality of cooked rice was associated with cuisine type (p < 0.00001), rice type (p < 0.01), cooking (p < 0.01), serving methods (p < 0.00001), and management food hygiene training (p < 0.01). Examination of chicken sandwiches found that most (75%; 335 of 449) were of satisfactory/acceptable microbiological quality and 114 (25%) were unsatisfactory. Acceptable microbiological quality of sandwiches was associated with sandwich bars that had hazard analysis in place (p < 0.05). Smaller businesses, as indicated by Local Authority Inspectors' Consumer at Risk scores, were more likely to have samples classified as unsatisfactory or unacceptable compared to larger businesses (p < 0.001). The majority (90%) of premises had hand-washing facilities accessible and available for use, although only over half (55%) were correctly used as judged by the sampling officer. Where the manager of the premises had received some form of food hygiene training, food safety procedures such as the hazard analysis system were more likely to be in place (p < 0.0001). PMID- 12564244 TI - Microbiological quality of drinking water from mobile food vendors. AB - A survey of the microbiological quality of drinking water from mobile food vendors in Cheshire found that 50% of the samples were unsatisfactory according to current regulations for bottled water. There was an inverse correlation between total viable counts and frequency with which the water container was cleaned. PMID- 12564245 TI - Collaborative investigation of an outbreak of Salmonella enterica serotype Newport in England and Wales in 2001 associated with ready-to-eat salad vegetables. AB - In June 2001, as part of a microbiological study of bagged, ready-to-eat salad products, Salmonella enterica serotype Newport was isolated from a sample of pre packed green salad distributed by a major supermarket retailer. The strain was characterised by phage typing, plasmid profile typing and pulsed-field gel electrophoresis. Other isolates of S. Newport from cases of human infection in England and Wales in the first six months of 2001 were similarly characterised. Of 60 strains from cases of human infection, 19 were found to be indistinguishable from that isolated from the salad product. This study highlights the benefits of an integrated approach to outbreak investigations, involving the various elements of the PHLS and the Food Standards Agency, and acknowledges the full co-operation of the retailer in ensuring the rapid withdrawal of the contaminated product. PMID- 12564246 TI - Accuracy of routine data on MMR vaccination coverage and validity of parental recall of vaccination. AB - A study was conducted to re-validate the measles, mumps and rubella (MMR) coverage and measles susceptibility in children resident in Bolton and born between 1991 and 1995; also to examine the accuracy of parental recall of children's MMR vaccination status. Using general practitioner/practice held records, a validation survey was conducted on a random sample of 171 reportedly unvaccinated children. A questionnaire survey was used to assess parental recall. 'Records' to 'recall' agreement was examined. One- and two-dose population coverage according to routine data was 94.6% and 75.2% respectively, with measles susceptibility of 8.1%. Based on the 'records' survey, one- and two-dose population coverage estimates were 97.4% (+2.8%) and 78.9% (+3.7%), with a revised susceptibility estimate of 5.2%, respresenting a 36% reduction in the susceptibility fraction. Routine data may modestly underestimate vaccination coverage and significantly overestimate measles susceptibility. Many parents of MMR unvaccinated children believe that their children have been vaccinated. PMID- 12564247 TI - Mumps virus reinfection--clinical findings and serological vagaries. AB - Sera taken from two cases of mumps reinfection were tested against different strains of mumps virus in a plaque reduction neutralisation assay. Antibodies to mumps in the sera reacted with all strains, but the pattern of emergence of these antibodies differed with the strains tested. These findings raise the possibility of emergence of a mutant strain under the selective pressure of immunisation, with limited or no cross protection induced by the vaccine strain. PMID- 12564248 TI - Susceptibility of young offenders to measles and rubella: an antibody prevalence study using oral fluid samples. AB - Three hundred and two oral fluid samples collected in January 2001 from male young offenders aged 18 to 21 years at HMP Hindley were screened for measles and rubella specific IgG, using 'in-house' amplified ELISA assays. MMR (measles/mumps/rubella) vaccine, offered at the same time, was accepted by 68.1% of the prisoners (92.7% of those agreeing to oral fluid antibody testing). Antibody prevalences were 92.1% for rubella and 80.8% for measles. This may, however, underestimate true prevalence as the sensitivity of the oral fluid assays used may be relatively poor in adults. Susceptibility levels are theoretically high enough to result in outbreaks should either infection enter this closed institutional environment. Since a measles outbreak would carry significant morbidity and rubella would represent a risk to associated non-immune women of childbearing age, it is suggested that prison health services should offer MMR to young offenders on entry. PMID- 12564249 TI - Universal hepatitis B vaccination of UK adolescents: a feasibility and acceptability study. AB - The feasibility of introducing universal hepatitis B immunisation was assessed by offering the vaccine to all 11-12 year old pupils in Greater Glasgow (approximately 10,800). Consent was received from 92% of the school roll, and 91.3%, 89.2% and 80.3% received at least 1, at least 2, and 3 doses respectively. The findings of this study constitute key evidence for the ongoing debate in the UK on hepatitis B vaccination. PMID- 12564250 TI - Potential of specialist drug services to deliver hepatitis B vaccination. AB - The city of Glasgow has a large population of injecting drug users (IDUs) who are likely to be susceptible to acute hepatitis B infection. We evaluated the effectiveness of a hepatitis B vaccination programme delivered at three existing types of services for IDUs. It was found that while uptake rates were generally low, they were higher at services where throughput of patients was lower, where clients had continuing relationships with services and where staff training and confidence were better. PMID- 12564251 TI - Comparison of conventional and accelerated hepatitis B immunisation schedules for homeless drug users. AB - This historical control study examines the uptake of two hepatitis B immunisation schedules at an inner city primary care centre for homeless people in Northern England. Originally homeless patients disclosing current or past illicit drug use were offered hepatitis B immunisation. In 1999 a conventional hepatitis B vaccine schedule was offered (immunisations at 0, 1, and 6 months) whereas in 2000 an accelerated schedule was introduced (immunisations at 0, 7 and 21 days). There was an increase in the uptake of hepatitis B vaccination by homeless drug users once the accelerated schedule was introduced. Furthermore, the completion rates for the accelerated vaccination regimen were almost seven times higher than for the conventional one. This indicates that the accelerated hepatitis B schedule should be the regime of choice for patients with a current or past history of drug use. PMID- 12564252 TI - Socioeconomic inequality and meningococcal disease. AB - Incidence of meningococcal disease was associated with socioeconomic deprivation across a rural English region. In young children the incidence was twice as high in the most deprived compared with the least deprived electoral wards. By addressing social inequalities the incidence of this serious infection could be reduced. PMID- 12564253 TI - PCR and the notification of meningococcal disease. AB - The polymerase chain reaction (PCR) can be used to investigate suspected meningococcal infection, with rapid results. We became aware that Birmingham Health Authority was not being informed about certain patients on whom meningococcal PCR had been performed. To establish whether notification was overlooked in some patients with suspected meningococcal disease, or whether PCR was performed on patients in whom it was thought meningococcal disease was unlikely, we compared numbers of patients notified to us over six months who had PCR with numbers of patients not notified who had PCR. Between 1st August 1999 and 31st January 2000, 54 PCR requests were made on seventy-seven notified patients; 51 PCR requests were made on non-notified patients and none were positive. Median age and length of stay were shorter in the non-notified patients. We conclude: (i) PCR may be being used inappropriately on younger patients who have shorter admissions and are unlikely to have meningococcal disease; (ii) in a number of patients initially admitted and notified with meningococcal disease the diagnosis is subsequently reconsidered. A 'denotification' procedure for such patients could make information on meningococcal disease more reliable. PMID- 12564254 TI - Tuberculosis in an indigenous young white south Wales population. AB - Concerns about tuberculosis in young people in two linked deprived non metropolitan communities triggered an investigation into local disease incidence and control measures. A pragmatic case-finding exercise, epidemiological enquiry and DNA genotyping were carried out to establish incidence and connections between cases over a five-year period. Of the 91 cases identified, 88 were from the white indigenous population. Approximately one third of cases were aged under thirty. Eight disease clusters were identified. Epidemiological investigation found spurious links between these, and DNA genotyping was necessary to clarify links. Seven cases and several contacts had been in prisons throughout the United Kingdom. Index cases under thirty generated more contacts, more contacts needing prophylaxis and more secondary cases than older patients. The workload resulting from tuberculosis control in younger cases was considerable. Tuberculosis remains an important problem in this population. PMID- 12564255 TI - Serial surveys of the tuberculosis nursing and support provision in the high incidence districts of England and Wales. AB - Postal surveys were carried out in 1998 and 2001 in the 43 high incidence TB districts of England and Wales (defined as an incidence of 15/100,000 per annum or greater). The provision of TB nursing and support staff was then compared with that recommended by the Joint Tuberculosis Committee for the numbers of TB notifications in 1997 and 2000 in the same areas. Forty-two districts replied in 1998, and all 43 replied in 2001. In 1998 only six out of the 22 high incidence areas outside Greater London met the minimum criteria for both nursing and clerical provision; none in Greater London did so. In 2001 only five out of 22 high incidence areas outside and one out of 21 inside Greater London met both criteria. The provision of nursing and clerical support for tuberculosis falls below minimum criteria in a substantial majority of high incidence areas and is not improving. Unless addressed, continuing under-provision of resources could itself contribute to a further rise in cases of tuberculosis. PMID- 12564256 TI - Assessment of the impact of BCG vaccination on tuberculosis incidence in south Asian adult immigrants. AB - Although BCG vaccination is recommended for tuberculin negative adult immigrants, there has hitherto been no evidence of its efficacy in this group. This epidemiological study compares the incidence of tuberculosis in a cohort of South Asian adult immigrants vaccinated on entry to the UK with its incidence in cohorts of immigrants of the same age and origin estimated from the five-yearly national surveys. Results suggest that BCG vaccine may have reduced the incidence of tuberculosis by up to 87.6% (95% confidence interval 55.2% to 98.5%) and support the current recommendation to vaccinate this high-risk group. PMID- 12564257 TI - Varicella vaccination--an ethical issue. PMID- 12564258 TI - Non-invasive diagnostics in man and other primates. PMID- 12564259 TI - The big bang--evolution of PHLS standard operating procedures. PMID- 12564260 TI - Chinese cultural influences on parental caregiving obligations toward children with disabilities. AB - The author draws on the personal and public dilemmas that result when Chinese families care for children with disabilities. She employed an ethnographic approach to interview 15 Hong Kong Chinese families caring for children aged 5 to 16 with disabilities. Twelve of these families included sons with disabilities. The following themes emerged: disruptions to natural order, public opinions on what constitutes personhood and ordered bodies, and the establishment of moral reputations linked to shame and blame and the gendered division of parenting. The author highlights the centrality of parenthood in Chinese society as the acceptance of responsibility and its cyclic enactment, and emphasizes how the care of children with handicaps strains and violates the Chinese culturally expected order of parental obligations. PMID- 12564261 TI - Older women's response to residential relocation: description of transition styles. AB - Older women relocating to institutional facilities are at risk for negative outcomes such as poorer health, decreased self-esteem, increased sense of social isolation and loneliness, loss of social support, and depression. However, little is known about relocation to congregate living facilities (CLF), the relocation transition process, or factors that contribute to outcomes. The perspectives of 31 women on their relocation experiences to CLF were elicited using a naturalistic approach. The Schumacher & Meleis nursing model of transitions was used to focus the study. Three distinct relocation transition styles emerged: Full Integration (45%), Partial Integration (42%), and Minimal Integration (13%). By understanding the characteristics and risk factors of relocation, unhealthy outcomes can be minimized and healthy outcomes promoted. PMID- 12564262 TI - Caring for patients under Medicaid mandatory managed care: perspectives of primary care physicians. AB - The purpose of the study was to describe the experiences of primary care physicians caring for Medicaid recipients in a demonstration mandatory health maintenance organization (HMO) managed care program. The authors collected data through semistructured individual or focus group interviews with 14 physicians and through interviews with the chief executive officers of the three HMOs participating in the demonstration program. Interview questions, developed initially from a review of the literature, addressed physicians' experiences as primary care providers for Medicaid recipients under traditional fee-for-service and under managed care arrangements through the demonstration program. Four themes emerged: providers' hassles and burdens, the complex needs of Medicaid patients, improved access to care under managed care, and individual providers' disconnect from the processes of health policy implementation and program evaluation. PMID- 12564263 TI - "This is a Spiritual Experience": perspectives of Latter-Day Saint families living with a child with disabilities. AB - The presence of a child with disabilities elicits a variety of stress demands on the family. Religion is recognized as a powerful personal, family, and cultural variable. However, little is known about the influence of religion in dealing with disability among families within particular religious groups. This descriptive study explored themes of spiritual belief and religious support among families of the Church of Jesus Christ of Latter-Day Saints (LDS, or Mormon) with a child with developmental disabilities. Parents shared perspectives of meaning that emerged from experiences with religion and family beliefs perceived to be unique. The core theme, "This is a Spiritual Experience," provides the foundation for a descriptive model that depicts aspects of finding meaning and perceived transcendence. PMID- 12564264 TI - Images of recovery: a photo-elicitation study on the hospital ward. AB - The authors attempted to discover the part the physical setting of the ward plays in patients' recovery by asking patients to take photographs of their ward, its spaces and objects, and then interviewing them about these images in hospital and subsequently in their homes. Photography allowed the patients to identify and capture aspects of the setting that they found salient and provided the photo elicitation material for the interviews. Based on these data, the authors present (a) a critical discussion of the use of photography as method and its implications for qualitative analysis, (b) an overview of the kinds of image taken with respect to the ward and the course of patients' recovery, and (c) a theoretical analysis, employing Walter Benjamin's use of the concept of mimesis, that understands recovery as a bodily act in response to the shock to the senses that hospitalization and surgery produce. PMID- 12564265 TI - Redefining parental identity: caregiving and schizophrenia. AB - When parents try to assume responsibility for an ill adult-child with schizophrenia, the law, mental health practitioners, and often the ill person reject their right to do so. Consequently, these parents regard themselves as disenfranchised, i.e., lacking the rights required to care properly for their loved ones. Redefining Parental Identity, a grounded theory of caregiving and schizophrenia, traces changes in a parent's identity and caregiving during the erratic course of the child's mental illness. Participants were a purposive sample of 29 parent caregivers from 19 families in British Columbia, Canada, caring for 20 adult children. This understanding of their experience will be helpful to parents of people with schizophrenia, professional practitioners, and those involved in mental health care reform. PMID- 12564266 TI - Continuing to conduct research in nursing homes despite controversial findings: reflections by a research scientist. AB - To illustrate the potential controversial nature of the research findings, the author first presents data from an ongoing study on the care of terminally ill nursing home residents, then responds to the question: "How can you go into nursing homes, find out all that you do, publish your findings, and continue to gain access to nursing homes?" Strategies used over the past 20 years to gain this access and to develop and maintain harmonious relationships during and after the data collection process are presented. The author emphasizes the importance of conducting research in settings that may generate controversial findings. Furthermore, she states that these findings should be presented to advocates and policy makers who are in a position to bring about changes that will improve the quality of care. PMID- 12564268 TI - Reconsidering reflexivity: introducing the case for intellectual entrepreneurship. AB - In this article, the author reconsiders reflexivity and attempts to examine some unresolved issues by drawing particular attention to the relationship between reflexivity and certain related phenomena/processes: the researcher's a priori knowledge, values, beliefs; empathy within qualitative research; the presence and influence of the researcher's tacit knowledge, and May's "magic" in method. Given the limitations of some reflexive activity identified in this article, the author introduces the case for greater intellectual entrepreneurship within the context of qualitative research. He suggests that excessive emphasis on reflexive activity might inhibit intellectual entrepreneurship. Wherein intellectual entrepreneurship implies a conscious and deliberate attempt on the part of academics to explore the world of ideas boldly; to take more risks in theory development and to move away from being timid researchers. PMID- 12564267 TI - Consent in oral history interviews: unique challenges. AB - The literature on oral history methods has increased over the past decade. Yet, the issues in gaining consent warrant specific attention. The process of gaining consent in oral history interviews has unique features and varies from accepted procedures in qualitative research. The authors discuss the legalities and ethics of oral history interviews and provide examples regarding the consent process from an oral history project that they conducted. The researchers conclude that despite its complexity, presenting the transcribed interview to the interviewee contributes in an important way to the ethical integrity of the interview process. PMID- 12564269 TI - The role of primary care in bioterrorism, epidemics and other major emergencies: failing to plan is planning to fail. PMID- 12564270 TI - Disfigurement--neglected in primary care? PMID- 12564271 TI - Influences on the variation in prevalence of type 2 diabetes between general practices: practice, patient or socioeconomic factors? AB - BACKGROUND: The prevalence of type 2 diabetes is known to vary between countries, districts and general practices. The influence of early detection and screening on the variation of prevalence between general practices has not previously been investigated. AIM: To test the hypothesis that the prevalence of type 2 diabetes is associated with awareness of and screening for diabetes within general practices and to explore other factors that may explain the variation in prevalence between practices. DESIGN OF STUDY: Cross-sectional study of general practices. SETTING: Forty-two general practices in Newcastle and North Tyneside; 20% random sample of patients with type 2 diabetes (n = 1056). METHOD: Factors thought to be associated with the variation of type 2 diabetes prevalence were collected from general practices through practice managers, medical records, and patient questionnaire. Pearson's correlation coefficient was used to quantify the association, and variables significant at the 5% level were entered into a multiple linear regression model. RESULTS: There was a wide inter-practice variation in age/sex standardised type 2 diabetes prevalence (range = 0.69% to 2.73%; P < 0.001). There was no significant association between the prevalence of type 2 diabetes and the proportion of patients detected outside primary care or the proportion of patients detected through screening, accounting for only 2% and 3% of the variation in type 2 prevalence between practices, respectively. The mean Townsend deprivation score accounted for 42% of the variation in type 2 diabetes prevalence between practices, with more deprived practices having a higher prevalence. CONCLUSION: This study suggests that socieconomic deprivation, rather than detection through screening or awareness of diabetes, accounts for much of the variation in prevalence of type 2 diabetes between practices. PMID- 12564272 TI - Randomised controlled trial of therapeutic massage in the management of stress. AB - BACKGROUND: Many patients presenting in general practice are suffering from stress. Approaches using complementary therapeies are gaining popularity but have not been systematically evaluated. AIM: To compare the effect of six sessions of therapeutic massage with the use of a relaxation tape on stress, as measured by The General Health Questionnaire (GHQ-30). DESIGN OF STUDY: Randomised controlled trial. SETTING: A stress reduction clinic in general practice. METHOD: There were three treatment groups. Patients in the first group received six sessions of therapeutic massage, which was carried out by a nurse trained in this technique. Patients in the second group were given six sessions using a relaxation tape in the surgery and those in the third group were given a relaxation tape to use at home. The main outcome measures were: the GHQ-30, the Adapted Well Being Index (AWBI); a sleep scale; general practitioner (GP) consultations for any reason in the six weeks before treatment, during treatment, and the six weeks following treatment; and patient satisfaction. RESULTS: Sixty-nine patients completed the treatment. There were significant improvements across all four outcome measures and all three treatment groups. Following treatment the majority of patients felt far less emotional disturbance, were sleeping better, and consulting their GP less. CONCLUSION: Despite very strong patient preference for therapeutic massage, it did not show any benefits over either a relaxation tape used in the surgery or a relaxation tape used at home. PMID- 12564273 TI - Predictors of hip joint replacement in new attenders in primary care with hip pain. AB - BACKGROUND: Studies investigating the factors associated with need for total hip replacement should ideally be based on prospective investigation of new attenders in primary care. AIM: To determine the incidence of listing for total hip replacement, and its predictors, among attenders in primary care with a new episode of hip pain. DESIGN OF STUDY: Prospective multicentre cohort study. SETTING: One hundred and ninety-five patients (mean age = 63 years, 68% female) with new episode of hip pain, attending primary care between November 1994 and October 1997. At the first visit, patients were evaluated for indices of pain and disability, range of hip movement, and radiographic changes of osteoarthritis. METHOD: General practitioner participants were recruited from the membership of the Primary Care Rheumatology Society to recruit all consecutive attenders with a new episode of hip pain. Annual follow-up was carried out to determine which patients were being 'put on a waiting list' for total hip replacement. RESULTS: Seven per cent of patients were put on a waiting list for total hip replacement within 12 months and 23% of patients within four years. At presentation, pain duration, pain severity, (including the need to use a stick) and restriction of internal rotation were the major clinical predictors of being put on a waiting list. Radiographic predictors of osteoarthritis performed similarly to the clinical measures. A simple scoring system based on both radiographic severity and two of the clinical measures was derived that identified groups at high likelihood of being put on a waiting list (sensitivity = 76%) with a low false positive rate (specificity = 95%). CONCLUSION: New primary care attenders with pain are frequently accepted for total hip replacement soon after their first attendance--a decision that can be predicted by simple clinical measures. PMID- 12564274 TI - The diagnostic value of macroscopic haematuria for the diagnosis of urological cancer in general practice. AB - BACKGROUND: The diagnostic value and the impact of some signs and symptoms in most diseases in primary care have only been studied incompletely. AIM: To assess the diagnostic value of macroscopic haematuria for the idagnosis of urological cancer (bladder, kidney) in a general practice setting, as well the influence of age, sex, and some additional signs and symptoms. DESIGN OF STUDY: Diagnostic study. SETTING: The study was performed in a sentinel station network of general practices in Belgium, covering almost 1% of the population. SUBJECTS: All patients attending their general practitioner and complaining of haematuria during 1993 and 1994 were included for the prospective part of the study. Every patient diagnosed with a urological cancer in this period was registered for the retrospective part. METHOD: Mean outcome measures of sensitivity, specificity, positive and negative predictive value, and positive and negative likelihood ratio were used to assess diagnostic value. RESULTS: Within the registration year 1993-1994, patient-doctor encounters, related to 83,890 patient-years, were registered. The positive predictive value (PPV) for urological cancer was 10.3% (95% CI = 7.6% to 13.7%). Sensitivity was 59.5% (95% CI = 50.4% to 60.1%). The PPV of patients aged over 60 years was 22.1% (95% CI = 15.8% to 30.1%) for men and 8.3% (95% CI = 3.4% to 17.9%) for women. In the age group 40 to 59 years, the PPV was 3.6% (95% CI = 0.6% to 13.4%) for men and 6.4% (95% CI = 1.7% to 18.6%) for women. In the prospective part of the study, no urological cancer was found in the age group under 40 years. CONCLUSION: Men older than 60 years of age with macroscopic haematuria have a high positive predictive value for urological cancer. In these patients, a thorough investigation is indicated. In patients over 40 years of age of either sex, referral or watchful waiting can be justified. PMID- 12564275 TI - A qualitative study of patients' perceptions of acute infective conjunctivitis. AB - BACKGROUND: Acute infective conjunctivitis is a self-limiting condition that commonly presents to primary care. Patients' understanding of conjunctivitis, their reasons for attendance, and their responses to different management strategies, are unknown. AIM: To explore patients' understanding of conjunctivitis and its management. DESIGN OF STUDY: Qualitative study using semi structured one-to-one interviews. SETTING: Three general practices in Hampshire and Wiltshire. METHOD: Twenty-five patients presenting with conjunctivitis at their general practices were interviewed. Main outcome measures were patients' perceptions of conjunctivities, their experience and knowledge of the disease, beliefs regarding treatment, and their responses to different management strategies and a patient information leaflet. RESULTS: Patients regarded conjunctivitis as a minor illness, although some considered it might become more serious if not treated. Nearly all were confident at recognising conjunctivitis. They stated a preference for not taking medication, but believed that conjunctivitis would not clear up without treatment. However, they were open to alternative management approaches; for example, the delayed prescription approach, because they trusted their general practitioners' (GPs') judgement. Once they were aware of the self-limiting nature of conjunctivitis, patients felt they would prefer to wait a few days to see if the condition improved before seeking medical advice, even if this resulted in a few more days of symptoms. CONCLUSION: Patients who attend their general practices with conjunctivitis present for treatment because they are not aware of its self-limiting nature. Providing patients with this information may enable patients, enhance self management, and reduce the use of topical antibiotics and the demand for urgent general practice appointments. PMID- 12564276 TI - Parents' beliefs and expectations when presenting with a febrile child at an out of-hours general practice clinic. AB - On the basis of structured interviews with 146 parents, this study describes why and when parents of acutely ill children seek the out-of-hours service, what actions they might have taken beforehand, and their expectations as to the outcome of the consultation. A total of 46% of the parents did not consider their child's condition to be serious, but 12% throught that their child was very ill. Parents sought medical advice because of what they perceived to be a lack of control of the condition (49%), fear of a serious disease (17%), and for symptom relief (34%). All except three parents expected there to be an examination of their child, and 79% expected an explanation or a diagnosis. Only 13% spontaneously mentioned that they expected a prescription. It is clinical and communicative skills that prevail in promoting successful consultations in this setting. PMID- 12564277 TI - Pain and subsequent mortality and cancer among women in the Royal College of General Practitioners Oral Contraception Study. AB - Recent research suggested associations between pain and subsequent all-cause and cancer-specific mortality. This study examined death and cancer development within six years of reporting pain, among women in the Royal College of General Practitioners Oral Contraception Study. We found no associations between 'any' or 'chronic' pain and subsequent all-cause mortality or cancer. We found a higher risk of death from respiratory disease among women reporting pain (adjusted odds ratio [AOR] = 2.5), a higher mortality among women reporting chronic chest pain (AOR = 1.75), and a higher risk of subsequent cancer among women reporting head or abdomen pain. Given the high prevalence of pain symptoms, these findings may be important, and warrant further research. PMID- 12564278 TI - Doctors' characteristics do not predict long-term glycaemic control in type 2 diabetic patients. AB - Glycaemic control in type 2 diabetic patients varies widely between general practitioners (GPs). To increase our understanding of this variation, linear random effects models were used to examine the predictive value of GP characteristics on the course of annual HbA1c measurements, in 688 newly diagnosed type 2 diabetic patients between one and five years after diabetes diagnosis. We found that characteristics of centrally supported GPs, such as interest in diabetes, experience, practice type, list size, and weekly working hours, did not predict their patients' glycaemic control. PMID- 12564279 TI - The general practitioner-patient consultation pattern as a tool for cancer diagnosis in general practice. AB - A case-control study design was used to examine consultation patterns during the three years leading up to the diagnosis of an internal malignancy, within the context of the registered patient list of a single large general practice at Winterton, North Lincolnshire. Using a combination of matching, consultation subclassification, and conditional logistic regression, account was taken of the major confounders affecting consultation rates. Generally, the odds of cancer rose in tandem with increases in the average time between new consultations. This trend was significant for all breast cancers (at the 5% level) and persisted after adjustment for occupation, smoking, and marital status (P = 0.03), as well as after the exclusion of patients identified by routine screening (P = 0.05). PMID- 12564281 TI - Teaching old dogs new tricks. PMID- 12564280 TI - Walk-in centres in primary care: a review of the international literature. AB - Nurse-led walk-in centres were first announced in April 1999. They represent a new development in unscheduled care provision in the United Kingdom (UK) National Health Service (NHS). By the end of 2000, 40 NHS walk-in centres had been opened, with further centres recently announced. This paper aims to review international experience with walk-in centres in primary and emergency care and identify relevant lessons for the UK. This study is a systematic review, with qualitative synthesis of relevant findings. Studies were identified from seven major bibliographic databases using a sensitive search strategy, and 244 relevant documents relating to walk-in or 'ambulatory care' centres were identified. Users of walk-in centres in other countries tend to be a relatively affluent population of working age, and a different population from those using conventional general practice services. Walk-in centres are used particularly when other health services are closed. The problems presented are mainly minor illnesses and minor injuries. People choose this form of care mainly for reasons of convenience, and satisfaction with the service is generally high. The very limited evidence available suggests that walk-in centres provide care of reasonable quality, but there is insufficient evidence to draw firm conclusions about the impact of walk in centres on other healthcare services or the costs of such care. Although a number of countries have had a long experience of walk-in centres, the lack of reliable evidence on many of the most important issues is notable. In the NHS, walk-in centres represent a radically innovative attempt to improve access to health care, but the limited research available does little to inform their development. Important questions that need to be addressed include whether walk in centres do improve access to care, for whom, and at what overall cost. PMID- 12564282 TI - Virtue ethics. PMID- 12564284 TI - Grief counselling and depression. PMID- 12564283 TI - New GP contract and European definition. PMID- 12564285 TI - Inaccurate, leaky sphygmomanometers are still common. PMID- 12564286 TI - Preventing falls in primary care. PMID- 12564287 TI - Advanced access. PMID- 12564288 TI - [The language of medicine]. AB - The idiom of physicians has evolved from Latin to the genetic code, in parallel with a change in the paradigm of disease. From the end of the 18th century, disease was no longer attributed to a disturbance of the body equilibrium, but to a localised abnormality--of organs, cells and eventually genes. A further step in this direction is the proposed creation of physician-engineers to replace broken parts in the human machinery. Such blind reductionism fails to offer a solution for somatisation, a scourge of modern western societies. Even organ specialists should be able to revert to the holistic notion of 'disequilibrium' to account for unexplained symptoms. For the medical profession, an education in the humanities is no less important than that in science. PMID- 12564289 TI - [Tuberculosis spondylodiscitis in three older patients]. AB - Three patients, two women aged 78 and 70 years, respectively, and a man aged 71 years, had back pain for months. Fever was absent. Blood parameters of infection were slightly or highly elevated. The diagnosis of spondylodiscitis was confirmed by MRI in all three patients. In one patient tuberculosis was confirmed by culture. One patient was from Turkey and the other two patients had been exposed professionally to tuberculosis. On two occasions, spondylodiscitis was complicated by compression of the spinal cord, and surgical intervention was necessary on one occasion because of neurological deficit. Initial treatment consisted of long-term bed rest and antituberculous therapy. All three patients recovered successfully. Tuberculous spondylodiscitis is a rare cause of back pain, but should be included in the differential diagnosis, particularly if there is an increased risk based on the medical history or if the patient comes from an endemic region or has an increased risk due to his or her profession. PMID- 12564290 TI - [More safety for patients requires confidentiality in the enquiry into medical errors]. AB - Recently, a seven-month-old baby died after a cardiac intervention and a medical error was presumed. An independent committee investigated this mishap after the confidentiality of the report was guaranteed. However, because of a judicial ruling, the report had to be made public, which caused great upheaval. To improve safety, error management requires confidential investigation to ensure co operation of all parties involved. There are two conflicting and irreconcilable aims: medical professionals want to establish what went wrong, legal professionals want to establish who is at fault. The position of investigative committees should be (legally) fortified with a guarantee of confidentiality and sound underlying principles of investigation should be delineated. PMID- 12564291 TI - [Diabetic encephalopathy: an underexposed complication of diabetes mellitus]. AB - Diabetes mellitus seems to be associated with gradually developing end-organ damage to the central nervous system. This relatively unknown complication of both diabetes type 1 and type 2 can be referred to as 'diabetic encephalopathy'. Measurable manifestations are electrophysiological and structural changes and limitations in the cognitive functioning. The mechanisms responsible for this diabetic encephalopathy are only partially known. Chronic metabolic and vascular changes seem to play an important role. The effects of diabetes on the brain are most distinct in the elderly. This may be the consequence of interactions between the mechanisms that underlie the ageing of the brain, dementia and the origin of diabetic complications. At present there are few leads for the targeted diagnostics and treatment of individual patients. PMID- 12564292 TI - [Atherosclerosis and inflammation: the role of C-reactive protein]. AB - C-reactive protein (CRP) is an acute phase protein, the serum concentration of which can increase up to a 1000-fold after the onset of a stimulus. It plays a role in the aspecific immune response to bacteria and fungi and the clearance of apoptotic cell material. Inflammation plays an important role in the aetiology and pathogenesis of atherosclerosis and plaque formation. Approximately fifty percent of acute coronary syndromes are the consequence of unstable plaques rupturing, followed by thrombus formation. A characteristic of these unstable plaques is an increase in inflammatory cells (macrophages and T lymphocytes). The serum concentration of CRP might reflect the amount of inflammation within atherosclerotic plaques and thus might provide an indirect measurement of the instability of the plaques. CRP could therefore have a predictive value for the occurrence of plaque rupture. Furthermore, there are indications that CRP itself is active in the inflammatory process. Prospective studies have shown that so called high-sensitivity CRP (hsCRP) measurements could be used as a tool for determining the risk for acute coronary syndromes. The inflammation-inhibiting characteristics of statins and acetylsalicylic acid (especially the reduction of the hsCRP level) might contribute to reducing the risk of plaque rupture. PMID- 12564293 TI - [Withdrawing the use of acetylsalicyclic acid prior to an operation usually not necessary]. AB - Acetylsalicylic acid (aspirin) induces an irreversible inactivation of cyclo oxygenase in blood platelets which lasts for the entire period that the platelets remain in the circulatory system, 7 to 10 days. In order to prevent excessive bleeding, patients presenting for surgery are asked to stop using aspirin 10 days before the procedure. Some studies have found that aspirin causes increased peri operative blood loss, whilst other studies have found that it does not. All effect studies found in Medline (January 1966-May 2002) on surgery and bleeding complications due to aspirin were analysed. The studies available were assessed for methodological quality and the results were summarised in an evidence table. No clinically relevant bleeding complications were reported for cardiovascular, vascular and orthopaedic surgery and epidural anaesthesia. Most studies reported an increase in clinically non-relevant bleeding induced by aspirin. The literature contains too little information on cataract surgery, dermatological surgery, gynaecological and abdominal surgery, ENT and dental surgery, urological surgery, lung biopsy and endoscopic biopsy. In those types of surgery in which even a minor bleeding leads to severe complications, e.g. neurosurgery, aspirin should be withdrawn 5-10 days in advance. Also in patients with coagulation disorders, aspirin should be withdrawn prior to the operation. There is no scientific evidence for the withdrawal of aspirin in all patients, 5-10 days prior to surgery. Indeed for heart patients in particular, the continued use of aspirin is recommended. PMID- 12564294 TI - [Diagnostic image (120). A neonate with fever and a rash. Gianotti-Crosti syndrome]. AB - A 14-month-old boy presented with fever and an itching papulous skin eruption. Based on the clinical presentation and the histopathologic findings in the skin biopsy, the diagnosis of Gianotti-Crosti syndrome was made. PMID- 12564295 TI - [Bladder control in 1-4 year old children in the the Eindhoven and Kempen region (The Netherlands) in 1996 and 1966]. AB - OBJECTIVE: To determine the age at which children gain bladder control and to compare this with the data from 30 years ago. DESIGN: Questionnaires. METHOD: On the basis of the findings of a 1966 study into toilet training in the Eindhoven and de Kempen region, the Netherlands, a questionnaire was drawn up and distributed via 30 child-health clinics in this region to parents of children aged 12-59 months, during the period 1 March-30 June 1996. The results were compared with those of the earlier study. RESULTS: Data from 1176 children could be evaluated (response rate: 65%). In 1996, the median age for bladder control in boys during the day was 32.6 months and 40.5 months for night-time control. In 1996 boys achieved daytime bladder control 6.7 months earlier and night-time control 7.2 months earlier. In 1996, the median age for girls was 29.7 months for daytime control and 35.4 months for night-time bladder control: in 1966 girls achieved daytime and night-time bladder control 8.2 and 4.8 months earlier, respectively. Factors associated with earlier bladder control were: early age at which parents started toilet training, presence of other children in the family, early age at which the child attended a day-care centre, early age at which the child was able to walk. Other factors such as the presence of a complete family set, parental level of education and professional situation did not show a correlation with the age at which the child achieved bladder control. The type of diaper used was an additional factor for bladder control at all ages but was only statistically significant for 3-year-olds, both during the day and during the night. CONCLUSION: Children in the Eindhoven region achieved daytime and night time bladder control at a significantly later age than 30 years ago. Various factors such as toilet-training age, day-care attendance, family size and type of diaper played a role in this phenomenon. PMID- 12564296 TI - [Dementia with Lewy bodies; 2 patients with exacerbation due to an atypical antipsychotic, but with a favorable response to the cholinesterase inhibitor rivastigmine]. AB - In two patients, men aged 80 and 75 years with cognitive deterioration, hallucinations and parkinsonism, the clinical diagnosis 'dementia with Lewy bodies' was established. Treatment with an atypical antipsychotic, risperidone and olanzapine respectively, resulted in an exacerbation of the parkinsonism. Rivastigmine evidently improved the psychosis, the anxiety and the cognitive, mood and behaviour disorders. Titrated treatment with levodopa improved the mobility without an increase of the psychosis. The treatment of dementia with Lewy bodies is complex. Levodopa can lead to an increase in visual hallucinations. Antipsychotics often cause serious side effects, such as increasing parkinsonism, sedation and cognitive deterioration. Cholinesterase inhibitors such as rivastigmine could possibly provide an alternative treatment for the neuropsychiatric symptoms. PMID- 12564297 TI - [Psychiatric side effects associated with diphosphonate treatment]. AB - Two women aged 50 and 64 years, respectively, and one man aged 43 years, were treated with cyclic etidronic acid for osteoporosis. After some months/years they developed mood, concentration and memory problems. The complaints diminished within several weeks after withdrawal of the drugs, and reappeared after rechallenge. Two of these patients had similar complaints during the use of another bisphosphonate (pamidronic acid and alendronic acid, respectively). Bisphosphonates are used increasingly frequently for the treatment of osteoporosis. Generally, these drugs are well tolerated. The most frequently reported adverse events are gastrointestinal complaints and oesophageal ulcers. Psychiatric complaints also appear to be a rare side effect. PMID- 12564298 TI - [Dealing with patient dissatisfaction; a professional skill]. PMID- 12564299 TI - [Dealing with patient dissatisfaction; a professional skill]. PMID- 12564300 TI - [Exteriorization of the colon with segmental resection (Mikulicz operation): history and current considerations]. PMID- 12564301 TI - [Photodynamic therapy in cardiac and vascular surgery]. AB - One of the major drawbacks to therapeutic cardiovascular intervention is intimal hyperplasia and constrictive remodeling, which result in vascular restenosis. Neointimal hyperplasia is characterized by proliferation and migration of smooth muscle cells. These cells also produce new extracellular matrix, leading to narrowing of vessels. Photodynamic therapy (PDT) represents one of the most promising approaches to the inhibition of intimal hyperplasia. PDT requires the interaction among 3 factors: a source of light, usually a laser, a photosensitizer and oxygen. When the inert photosensitizer absorbs light of a specific wavelength, it is activated to an excited triplet state, generating reactive oxygen species. These free radicals are able to induce apoptosis of the smooth muscle cells that had absorbed the photosensitizer; they also induce changes in the extracellular matrix, reducing cell migration. Because of continued success of PDT in inhibiting intimal hyperplasia in experimental animal models, it is now being tested in clinical trials for vascular diseases. PDT offers many advantages to the surgeon since it can act on numerous factors responsible for vascular lesions. In the future PDT could be used in helping to overcome the inherent failures associated to vascular reconstruction. This treatment modality is emerging as an encouraging therapeutic option, either alone or as an adjunct to conventional treatment. However, more detailed clinical investigation are necessary to determine its full potential. PMID- 12564302 TI - [Biliary ileus. Considerations in surgical strategy]. AB - Gallstone ileus is the small bowel obstruction due to one or more biliary calculi stopped in the enteric lumen. This older patient pathology is often associated with other comorbid medical conditions. Two clinical reports are analyzed and the various surgical procedures critically evaluated. PMID- 12564303 TI - [Anastomotic dehiscence in colorectal surgery]. AB - Anastomotic leakage represents the most dangerous complication in digestive surgery and, particularly, in colorectal one. Indeed a careful evaluation and modification of all possible risk factors and a correct procedure aren't often enough to overcome a potential postoperative leakage or at least material filtration. The Authors review their series to analyse the role of all possible general and local risk factors and to verify what conditions can suggest an two or three stages operation. PMID- 12564304 TI - [Cystadenocarcinoma in situ of the urachus: a clinical case]. AB - Urachal carcinoma is a rare neoplasm which represents 0.01% cancers in adults. If this neoplasm is unusual, the development of cystadenocarcinoma on a urachal cyst is still more rare. The surgical treatment of choice is en-bloc excision of umbilicus and median umbilical ligament with partial cystectomy. As the urachal cystadenocarcinoma of Authors' patient was in situ and without bladder-invasion, they performed a more conservative treatment without partial cystectomy. They believe that this treatment may be indicated in association with a close follow up after surgery. PMID- 12564305 TI - [Surgical treatment of acute necrotic-hemorrhagic pancreatitis]. AB - The clinical course of an episode of acute pancreatitis varies from a mild transitory form to a severe necrotizing form characterized by multisystem organ failure and mortality in 20% to 40% of cases. Mild pancreatitis does not need specialized treatment, and surgery is necessary only to treat underlying mechanical factors such as gallstones or tumours of papilla of Vater. On the other hand, patients with severe necrotizing pancreatitis need to be identified as early as possible after the onset of symptoms to start intensive care treatment. Patients with infected necrosis, approximately 10 to 20%, must undergo surgical intervention, which consists of an organ-preserving necrosectomy followed by postoperative drainage-lavage or the method of "open abdomen". The Authors report a series of 20 patients operated on for acute necrotizing pancreatitis from 1998 to 2000. They describe the indications and methods of treatment, in particular the additional procedure following necrosectomy. PMID- 12564306 TI - [Superior vena cava syndrome]. AB - Superior vena cava syndrome is due to an intrinsic or extrinsic caval obstruction that evolves in acute or subacute way with distinctive clinical feature such as respiratory symptoms and venous stasis. Since 1998 we have treated three cases of spontaneous superior vena cava thrombosis in neoplastic patients who underwent several infusion of chemotherapy, respectively for a breast, uterine and rectum cancer. All patients was female, 52, 58 and 70 years old. The first two cases was treated with locoregional thrombolysis by infusing Urokinase 50,000 UI/h during 24 hours and Urokinase 50,000 UI/h during 12 h the third one. After that, we have positioned a 16/9 wallstent: in the first two cases directly into the superior vena cava, in the third case in the subclavian-anonyma truncus. We had in all cases the complete opening of the stent within the first 48 hours without complications, enabling us to reach a free caval diameter of about 2 cm with resolution of the clinical signs. In patients with high surgical risk, the caval wall-stent is the first choice to solve the vein recanalization. PMID- 12564307 TI - [Carcinoma of the male breast. Prognostic factors and outcome of surgical treatment]. AB - A retrospective study was made on 18 male patients with breast carcinoma treated at the Department of Surgery "Pietro Valdoni" of the University "La Sapienza" of Rome, Medical School. Demographics, pathology, stages, and treatment were determined from clinical reports. All patients but one underwent modified radical mastectomy. The length of follow up averaged 57.5 months. Five years actuarial survival rate was 62%. In the current study the Authors suggest that the clinical, prognostic and treatment features of breast carcinoma in men are similar to those reported in literature for post-menopausal women. PMID- 12564308 TI - Spontaneous rectus sheath haematoma in HCV mixed cryoglobulinemia requiring emergency treatment (case report). AB - Hepatitis C virus-related mixed cryoglobulinemia is a vasculitis, associated with a number of systemic manifestations and diseases. The Authors present a patient with mixed cryoglobulinemia, who developed acute abdominal pain and severe anemia, caused by spontaneous haematoma of rectus abdominis sheath. Prompt diagnosis of this unusual complication of cryoglobulinemia vasculitis may allow conservative treatment and avoid surgery, which is more expensive and invasive. To the best of Author's knowledge, spontaneous rectus sheath haematoma in a patient with cryoglobulinemia has not been previously reported. PMID- 12564309 TI - [Cutaneous Merkel cell carcinoma: case report]. AB - Merkel cell carcinoma is an unusual cutaneous malignancy with a propensity for spreading to regional lymph nodes, with recurrence at original site or/and in lymph node. Occurring most often on the head and neck of the elderly patients. Complete surgical resection is the mainstay of treatment of the primary tumor. A case is here described of an old patient with more than one local relapse and in regional lymph nodes too, complete surgical resected, actually in good state of health, treated with radiotherapy. PMID- 12564310 TI - [Gastrointestinal stromal tumors]. AB - The Authors report two cases of GIST and underline cytogenetic, clinic and prognostic aspects of this disease; they describe as well new histo-cyto-genetic data that identify as origin tissue interstitial cells of Cajal, by immunohistochemical and genetic tests. They also consider new diagnostic and therapeutic options based on surgical treatment. Finally controversial prognostic problems are discussed. PMID- 12564311 TI - [Surgical resection of hepatic hilar tumors]. AB - Hepatic hilar cancer has an extremely poor prognosis and resection for cure is a realistic possibility in only 15-20% of patients. Tumours confined strictly to the biliary confluence can often be excised locally without resorting to hepatic resection (Bismuth's type I, II). Tumours extending beyond the second order bifurcation (Bismuth's type III) require hepatic resection. In the period 1996 1998 ten patients with hilar cancer (adenocarcinoma) underwent curative resection at our Institution. There were 9 men and 1 women with a mean age of 61.7 years (range 49-76 yrs). One neoplastic lesion was Bismuth's type I, five type II, four type III. The mean preoperative bilirubin level was 20 mg% and the mean duration of jaundice was 4 weeks. Four patients had skeletonization resection of the tumour and extrahepatic bile ducts, clearing all lymphocellular and other tissue from the hepatic pedicle and coeliac axis. Bilioenteric continuity was reestablished by a Roux-en-Y jejunal loop with separate biliary duct anastomoses. Six patients required also hepatic resection to adequately remove the tumour (1 right hepatectomy, 2 right lobectomy, 2 left hepatectomy, 1 segmentectomy III). Three patients had liver metastases. One patient had involvement of the left arterial and portal branch. The postoperative staging was 2 stage II, 1 stage III, 7 stage IV. In 5 patients hepatic lymph nodes (N1) were involved. In no patient the tumour was found at the margin of resection. The median estimated blood loss for hepatic resection was 1,000 ml and for skeletonization 500 ml. Intraoperative mortality was 0%. Operative mortality was 20%. Three patients had a complicated postoperative course (1 cerebral TIA, 1 multiorgan failure, 1 ictus cerebri). All patients died. The mean postoperative survival was 7.4 months. Four patients (N1+) died of local tumour recurrence at 8, 11, 6, and 8 months. In our experience resective procedures can achieve a longer survival and a better quality of life. The operative mortality may be kept to a minimum by adequate selection of patients and technical expertise. PMID- 12564313 TI - Ophthalmological findings in a patient with mucolipidosis III (pseudo-hurler polydystrophy). A case report. AB - Mucolipidosis III (Pseudo-Hurler Polydystrophy) is a rare autosomal recessively inherited Hurler-like disease. The ophthalmological findings in these patients include a triad of mild retinopathy, corneal clouding and hyperopic astigmatism. We present a patient with these ophthalmological characteristics. PMID- 12564312 TI - [Optic neuropathy in Strumpell-Lorrain disease: presentation of a clinical case and literature review]. AB - This report describes a 36-year-old female with bilateral optic atrophy. Physical examination was normal except for subclinical signs of lower limbs spasticity. Her father was affected of Strumpell-Lorrain disease. The clinical characteristics of Hereditary Spastic Paraplegia or Strumpell-Lorrain disease are: progressive spasticity of the lower limbs and pathological reflexes. Pathologically, this disease is characterized by a degeneration or demyelinization of the cortico-spinal system and, to a lesser extent, of the posterior cord and the spino-cerebellar system. Optic neuropathy and dyschromatopsy have already been reported in a small group of patients. Visual acuity ranged from 20/20 to 20/200. Visual field alterations consisted of superior bitemporal amputation. Fundus examination showed optic atrophy. The incidence of optic neuropathy in Strumpell-Lorrain disease is quite high but its presence is often overlooked. Most patients are asymptomatic, but signs of optic nerve dysfunction are evident at clinical examination. According to some authors, there is an inverse relationship between the impairment of walking and the visual dysfunction. This was also noted in our case. PMID- 12564314 TI - [Ocular relapse of acute lymphoblastic leukemia]. AB - A bilateral leukemic hypopyon can be inaugural in the child's leukemia or reveal a relapse. A five years old child with acute lymphoblastic leukemia presented after 30 months of treatment a bilateral hypopyon. Anterior chamber paracentesis with cytological survey revealed leukemic cells and confirmed the ocular relapse. The treatment included the association of topical corticosteroids, chemotherapy and radiotherapy. This child died unfortunately 16 months later following a medullar relapse. We remind the different clinical aspects of leukemic invasion of the anterior segment and the therapeutic methods for this relapse. PMID- 12564315 TI - Diplopia as an initial manifestation of disseminated non-hodgkin's lymphoma. AB - A 37-year-old male presented a sudden diplopia. Ocular examination showed a partial paresis of the left eye (LE) on the left gaze, progressing in a few days towards a total ophthalmoplegia. Further investigation revealed an intra-orbital mass, immunohistologically diagnosed as a Diffuse Large B-cell Lymphoma (DLBCL), according to the WHO classification. Since the patient was in an advanced, disseminated stage of the disease (IVA-E), treatment was based on systemic and intrathecal chemotherapy with a pancranial radiotherapy. The clinical course was poor with only a 10-month survival. We wish to stress that the possibility of orbital malignancy in young adults with acute onset of ophthalmoplegia should be included in the differential diagnosis. PMID- 12564316 TI - Cat scratch disease (CSD) in patients with stellate neuroretinitis: 3 cases. AB - This case series describes three patients with a similar clinical picture: unilateral abrupt visual loss, optic nerve edema, and a macular star exudate. In all cases we found significant antibody titers to Bartonella henselae, the causative agent of cat scratch disease. Cat scratch disease seems to be the most common cause of stellate neuroretinitis, formerly known as Leber's idiopathic stellate retinopathy. A review of the pertinent literature shows that serologic evidence of B. henselae is sufficient to confirm the diagnosis given the low incidence of significant titers in the general population. Cat scratch disease is usually a self limiting disorder in immunocompetent patients, but treatment with doxycycline is recommended. PMID- 12564317 TI - Short-term intra-individual variability in heidelberg retina tomograph II. AB - We determined the short-term intra-individual variability for each parameter of the Heidelberg Retina Tomograph II (HRT II). Therefore we examined prospectively 20 healthy volunteers 3 times within 2 weeks. The variability was expressed as the coefficient of variation for each parameter. The short-term intra-individual variability in normal subjects was < or = 12% in all but 3 parameters. Rim Area was the least variable parameter (2 +/- 1%). Cup Volume had the highest variability (25 +/- 38%). PMID- 12564318 TI - Metastatic uveal melanoma: diagnosis and treatment. A literature review. AB - In the past three decades many efforts were done to improve the visual prognosis of patients with uveal melanoma. However, mortality has remained unchanged. The systemic prognosis depends on the size and other characteristics of the lesion. It is not affected by the choice of local treatment. This suggests dissemination at an early stage. The failure to improve survival is caused by difficulties in early detection of metastases and the limited susceptibility of these metastases to systemic therapies. PMID- 12564319 TI - Kickbacks in relation to medical practice. PMID- 12564320 TI - Child rape. PMID- 12564321 TI - Mass hysteria with possible pseudoseizures at a primary school. PMID- 12564322 TI - IUCD insertion at caesarean section--a new look. PMID- 12564323 TI - Frances Ames--human rights champion. PMID- 12564324 TI - Still getting community service right. PMID- 12564325 TI - Reaching for a fairer health care deal. PMID- 12564326 TI - Local hospital solution lasts longer. PMID- 12564327 TI - Challenges to cervical cancer screening in the Western Cape province. PMID- 12564328 TI - Early visual recovery after macular hole surgery with gas tamponade. PMID- 12564329 TI - Dissociative fugue. PMID- 12564330 TI - Rape survivors and the right to emergency medical treatment to prevent HIV infection. PMID- 12564331 TI - The epidemic of childhood obesity. PMID- 12564332 TI - Are low aminoglycoside doses appropriate when used for perioperative prophylaxis in urology? PMID- 12564333 TI - The role of nutrient intake in the aetiology of diabetes mellitus. PMID- 12564334 TI - The ART of rationing--the need for a new approach to rationing health interventions. AB - A key element in dealing with HIV/AIDS in South Africa depends on the resolution of the antiretroviral therapy (ART) paradox: while a universal First-World-style ART programme is unaffordable, a rationed treatment programme that includes ART is not only affordable but also vital for basic human rights reasons, to enhance prevention efforts and to keep the fabric of society together. Our recent paper on ART demonstrated how such a rationed programme would be both affordable and highly cost-effective. Traditional rationing mechanisms are unable to provide sufficient guidance as to how to go about this novel form of rationing. An alternative rationing mechanism is therefore proposed which seeks to balance ART in terms of three primary dimensions: total resource allocation to treatment, design of the treatment intervention, and setting targets on numbers to treat. Two secondary dimensions, related to total HIV and social spending, deserve equal attention. The current global context that precipitates and exacerbates the parallel contouring of disease burden and poverty should be constantly challenged. PMID- 12564335 TI - Women doctors in medical professional organisations in South Africa--a report by the Women in Medicine workgroup. AB - OBJECTIVES: To determine the level of representation of women doctors in medical professional organisations in South Africa historically and currently, and if not adequate, to ascertain the reasons for women's limited participation in medico political activities. DESIGN: A descriptive study of the membership of South African medical professional organisations and their executive structures. OUTCOME MEASURES: The number and percentages of women doctors as members and at executive level in medical professional organisations compared with the number of registered women practitioners. RESULTS AND CONCLUSIONS: Women are inadequately represented at all levels in the great majority of organisations for which information was provided. In view of the rapidly increasing number of women doctors the profession and its professional bodies need to take active steps to promote the participation of women in these organisations. PMID- 12564336 TI - Congenital eunuchism and Favorinus. AB - Ancient Hebrew literature as well as the New Testament differentiate between castrated eunuchs and congenital eunuchs. Congenital eunuchism is very rare today, and assuming that this was also the case in classical times, we investigated possible reasons why congenital eunuchs feature prominently. We discuss the probability that the concept 'congenital eunuchism' might in ancient times have included effeminate men who, according to cultural views on 'maleness' and androgyny, were almost equated with eunuchs. The causes of congenital hypogonadism are reviewed in order to attempt clarification of the condition of Favorinus, a congenital eunuch in the second century AD. We suggest that although he might have been a true hermaphrodite, as suggested by some authors, it is more likely that he had one of the following conditions: functional prepubertal castrate syndrome, testicular gonadotrophin insensitivity, selective gonadotrophin deficiency or Reifenstein's syndrome. PMID- 12564337 TI - Decommissioning & environmental restoration. PMID- 12564338 TI - Interview with Ninni Jacob. PMID- 12564339 TI - Hanford single-pass reactor fuel storage basin demolition. AB - The Environmental Restoration Contractor at the Hanford Site is tasked with removing auxiliary reactor structures and leaving the remaining concrete structure surrounding each reactor core. This is referred to as Interim Safe Storage. Part of placing the F Reactor into Interim Safe Storage is the demolition of the fuel storage basin, which was deactivated in 1970 by placing debris material into the basin prior to back filling with soil. Besides the debris material (wooden floor decking, handrails, and monorail pieces), the fuel storage basin contents included the possibility of spent nuclear fuel, fuel buckets, fuel spacers, process tubes, and tongs. Demolition of the fuel storage basin offered many unique radiological control challenges and innovative approaches to demolition. This paper describes how the total effective dose equivalent and contamination were controlled, how the use of a remote operated excavator was employed to remove high-dose-rate material, and how wireless technology was used to monitor changing radiological conditions. PMID- 12564340 TI - Cost-benefit of final radiological survey versus radioactive waste disposal. AB - A significant portion of the cost for D&D is related to low-level waste disposal, decontamination, and final radiological survey. For that reason, a careful cost benefit analysis must be performed weighing the cost of decontaminating and radiologically surveying the building media for release against the cost of disposing of the material as radioactive waste. This cost-benefit analysis visibly came into play at the Rocky Flats Environmental Technology Site during the recent D&D of two facilities. PMID- 12564341 TI - Fort St. Vrain--a DOE success story. AB - The Fort St. Vrain Nuclear Generating Station was a one-of-its-kind High Temperature Gas Cooled Reactor located near Denver, CO. Due to operational difficulties, the reactor was permanently shut down in August 1989. Plants to ship the spent reactor fuel to Idaho were thwarted by the Governor of Idaho. The inability to ship the spent reactor fuel offsite necessitated the construction of an Independent Spent Fuel Storage Installation (ISFSI) in order to proceed with the decommissioning of the reactor. After loading the ISFSI, decommissioning of the reactor was initiated and successfully completed. At the same time, the site was repowered using natural gas. As part of a settlement with the owner of the reactor, the U.S. Department of Energy agreed to take title to the spent fuel in the ISFSI and obtain an NRC license for the facility. The license for the ISFSI was transferred to the DOE in June 1999. Day-to-day operations of the ISFSI are accomplished by the DOE's Maintenance and Operations contractor, Bechtel Babcock Wilcox Idaho (BBWI). BBWI also operates the Three Mile Island Unit 2 ISFSI in Idaho for the DOE. This paradigm for ISFSI operations has been highly successful and is expandable to additional ISFSIs that may come under the jurisdiction of the DOE. PMID- 12564342 TI - Advanced characterization system deployments at the hanford site. AB - The Advanced Characterization System (ACS) uses readily available commercial radiation Environmental Restoration Contractor (ERC) detection and measurement equipment in a complementary staged system to locate and identity contamination in structures, systems and land area at Hanford's Environmental Restoration Contractor work sites. Large structures, such as former plutonium production reactors and separations facilities, are thoroughly and comprehensively characterized, not only for decontamination and decommissioning (D&D), but also for disposal and/or release. The ACS uses gamma-ray imaging instruments, in situ gamma spectrometry and large-area data-logging contamination survey meters to generate sufficient quality data to make operational and release decisions. The system is also used to safely characterize in-place systems without violating any containment and to localize activity in unusual situations. PMID- 12564343 TI - Effective radiological contamination control and monitoring techniques in high alpha environments. AB - In the decommissioning of a highly contaminated alpha environment, such as the one at Hanford's 233-S Plutonium Concentration Facility, one of the key elements of a successful radiological control program is an integrated safety approach. This approach begins with the job-planning phase where the scope of the work is described. This is followed by a brainstorming session involving engineering and craft to identify how to perform the work in a logical sequence of events. Once the brainstorming session is over, a Job Hazard Analysis is performed to identify any potential problems. Mockups are utilized to enable the craft to get hands on experience and provide feedback and ideas to make the job run smoother. Ideas and experience gained during mockups are incorporated into the task instruction. To assure appropriate data are used in planning and executing the job, our principal evaluation tools included lapel and workplace air sampling, plus continuous air monitors and frequent surveys to effectively monitor job progress. In this highly contaminated alpha environment, with contamination levels ranging from 0.3 Bq cm 2 to approximately 100,000 Bq cm-2 (2,000 dpm per 100 cm2 to approximately 600 million dpm per 100 cm2), with average working levels of 1,600-3,200 Bq cm-2 (10 20 million dpm per 100 cm2) without concomitant ambient radiation levels, control of the spread of contamination is key to keeping airborne levels As Low As Reasonably Achievable. PMID- 12564344 TI - Worker and environmental protection issues in the remediation of an abandoned source manufacturing facility. AB - The Gulf Nuclear Superfund Site located in Odessa, Texas, was an abandoned radioactive source production facility slated for cleanup as a Removal Action under the U.S. Environmental Protection Agency Region VI Superfund program. Prior to cessation of operations and abandonment of the facility in 1992, it was used for the production of radioactive sources used in the oil and gas industry and nuclear medicine applications. Pangea Group was contracted by the U.S. Army Corps of Engineers (USACE) Kansas City District to perform remediation of the site and other contaminated debris, cleaning of interior building surfaces, building demolition, and excavation/removal of contaminated soils and septic system. The project scope also included loading, containerization and transportation of low level radioactive wastes for offsite disposal. Primary radionuclides present at the facility were 137Cs, 60Co, and 241Am. The project also included packaging and removal of radioactive sources and mixed waste consisting of radiologically contaminated lead shot and lead source containers. Included in the paper is a discussion of primary worker protection and environmental protection measures employed on the project. Worker protection issues included the control of industrial and construction safety hazards as well as control of external and internal radiation dose. Control of air emissions and contaminated wastewater were also very important, especially due to the location of the site. The site was located in an area containing both residential and commercial properties. Several residences and businesses were located immediately adjacent to the site. The project involved the participation of the USACE Kansas City District, EPA Region 6, and the Texas Bureau of Radiological Health. Field work on the project started in April 2001 and was completed approximately five months later. PMID- 12564345 TI - Development of dose-based release limits for unrestricted release of a radiochemistry laboratory. AB - Current regulations for unrestricted release are based on annual dose equivalent. Unless one desires to use very conservative "screening levels," dose modeling must be accomplished to derive an areal or volumetric limit or concentration value for release purposes. Such derived limits are referred to as "Derived Concentration Guideline Levels" (DCGL). This paper describes the process employed to derive DCGLs for building surfaces contaminated with uranium and its decay progeny based on annual dose equivalent and the innovative means employed during the derivation. PMID- 12564346 TI - Remediation of Hanford's N-reactor liquid waste disposal sites. AB - Hanford's N-Reactor operated from 1963 to 1987 generating approximately 9 x 10(7) m3 of radioactive and hazardous liquid effluent as a result of reactor operations. Two liquid waste disposal sites, essentially large trenches designed to filter contaminants from the water as it percolates through the soil column, were established to dispose of the effluent. The discharges to the sites included cooling water from the reactor primary, spent fuel storage, and periphery systems, along with miscellaneous drainage from reactor support facilities. Today, both sites are classified as Treatment Storage and Disposal Facilities under the Resource Conservation and Recovery Act of 1976, which makes them priority sites for remediation. The two sites cover approximately 4,100 m2 and 9,300 m2, respectively. Remediation of the sites requires removing a combined total of approximately 2.6 x 10(8) kg of contaminated soil and debris. Principal radionuclides contained in the soil/debris are 60Co, 137Cs, 239Pu, and 90Sr. Remediation of these waste sites requires demolishing concrete structures and excavating, hauling, and disposing of contaminated soils in work areas containing high levels of contamination and whole body dose rates in excess of 1 mSv h-1. The work presents unique radiological control challenges, such as minimizing external dose to workers in a constantly changing outdoor work environment, maintaining contamination control during removal of a water distribution trough filled with highly contaminated sludge, and minimizing outdoor airborne contamination during size reduction of highly contaminated pipelines. Through innovative approaches to dose reduction and contamination control, Hanford's Environmental Restoration Contractor has met the challenge, completing the first phase on schedule and with a total project exposure below the goal of 0.1 person Sv. PMID- 12564347 TI - Ginger as an antiemetic during pregnancy. PMID- 12564348 TI - Therapeutic touch: a nursing practice. PMID- 12564349 TI - Therapeutic touch at the crossroads: observations on the Rosa study. PMID- 12564350 TI - Outcomes of touch therapies during bone marrow transplant. AB - CONTEXT: The integration of complementary modalities into mainstream healthcare is gaining increasing emphasis. It is important, therefore, to document the effects of these interventions on patient outcomes. OBJECTIVE: To investigate the effects of Therapeutic Touch and massage therapy on the outcomes of engraftment time, complications, and perceived benefits of therapy during bone marrow transplant. DESIGN: Randomized clinical trial. PARTICIPANTS/SETTING: Subjects were adult patients on the bone marrow transplant unit of a large urban tertiary care center. INTERVENTIONS: Subjects were randomly assigned to 1 of 3 treatment groups: Therapeutic Touch (TT), massage therapy (MT), and a control group called the friendly visit (FV). Subjects (N = 88) were stratified by type of transplant (allogeneic or autologous). Twenty-seven subjects received MT; 31 received TT; and 30 received FV. Nurses with expertise in the 2 touch therapies administered them. The interventions of MT, TT, and FV were administered according to standarized protocols every third day beginning the day chemotherapy began until discharge from the program. OUTCOME MEASURES: Time for engraftment, complications, and patient perceptions of benefits of therapy were the main outcome measures. Analysis of variance and analysis of covariance were used to determine significant differences among the 3 groups with respect to time of engraftment. RESULTS: A significantly lower score for central nervous system or neurological complications was noted for subjects who received MT comppared with the control group; however, no differences were found among the 3 groups with respect to the other 10 complication categories or in the total mean score for complications. Patients' perception of the benefits of therapy (total score) was significantly higher for those who received MT compared with the FV control group. The mean scores on the comfort subscale were significantly higher for patients receiving both MT and TT compared with the FV control group. CONCLUSIONS: Massage therapy may be effective in altering the psychological and neurological complications associated with chemotherapy during bone marrow transplant. Both massage and Therapeutic Touch provide comfort to patients undergoing this challenging process. PMID- 12564351 TI - The effects of back massage before diagnostic cardiac catheterization. AB - CONTEXT: Admission to the hospital for a diagnostic cardiac catheterization can be perceived as a threat to one's health status. Autonomic nervous system arousal, particularly the sympathetic division, can elicit negative physiological and psychological human responses as a reaction to this threat. OBJECTIVE: The purpose of this study was to measure the effects of a 20-minute back massage on the physiological and psychological human responses of patients admitted for a diagnostic cardiac catheterization. DESIGN: A randomized clinical trial design was used. Data were compared in a repeated measures design before massage (T1), immediately following the back massage or standard care (T2), and 10 minutes later (T3). SETTING: A large urban academic medical center. PARTICIPANTS: Forty six subjects admitted from home for a diagnostic cardiac catheterization. MAIN OUTCOME MEASURES: Heart rate, heart rate variability, blood pressure, respiration, peripheral skin temperature, pain perception, and psychological state. INTERVENTION: A 20-minute back massage. RESULTS: There was a significant difference between subject effect for group, with a reduction in systolic blood pressure in the treatment group (F = 8.6, P < .05). In addition, main effects were noted for time for diastolic blood pressure (F = 5.44; P < .006), respiration (F = 10.6; P < .005), total Profile of Mood States score (F = 5.9; P < .001) and pain perception (F = 4.09; P < .04) in both groups. CONCLUSIONS: A 20 minute back massage appeared to reduce systolic blood pressure in patients awaiting a diagnostic cardiac catheterization, while preparatory time in the cardiac catheterization laboratory appeared to reduce diastolic blood pressure, respiration, perceived psychological distress, and pain. PMID- 12564352 TI - A nurse-statistician reanalyzes data from the Rosa therapeutic touch study. AB - This article presents a reanalysis of data used to support the work of Emily Rosa's Therapeutic Touch (TT) science fair project published as an article in the Journal of the American Medical Association (JAMA) in 1998. The purpose of this article is to take a closer look at the assumptions, data, statistical procedures, and conclusions of the JAMA article. This is accomplished by focusing on (1) the conclusion that there was no overall effect of TT, (2) the conclusion that TT practitioners did not perform better depending on which hand was used, and (3) the assumptions about the capability of Rosa's experiment to validate an existing skill. Reanalysis of the Rosa data suggests contradictions to the authors' conclusions. Based on this reanalysis, the authors' recommendations against the use of TT can and should be challenged because of inappropriate design and analysis as well as incorrect statistical assumptions and conclusions. PMID- 12564353 TI - Assessing the consistency of traditional Chinese medical diagnosis: an integrative approach. AB - Traditional Chinese medicine (TCM) differs from its Western counterpart in a number of ways. One of the more striking ways involves a conceptually oriented diagnostic system that relies more on the clinician's reading of the patient's symptoms and signs than on laboratory findings. Because highly individualized TCM treatment plans emanate directly from its diagnostic system, a necessary condition for conducting clinically relevant TCM efficacy trials rests on the answer to a simple question: How consistent are different TCM practitioners at making the same TCM diagnoses and prescribing comparable treatment regimens for the same group of patients? Unfortunately, this question has not been adequately investigated, nor has the related question: Could the TCM diagnostic process be enhanced by access to modern biomedical tests? The purpose of this paper, therefore, is to discuss a number of the conceptual and methodological issues involved in the design of a recently funded NIH study whose primary purpose is to address these 2 questions. PMID- 12564355 TI - Robert Thurman, PhD gifts from Tibetan medicine. Interview by Bonnie Horrigan. PMID- 12564354 TI - Is acupuncture safe? A systematic review of case reports. AB - OBJECTIVE: The greater acceptance of acupuncture by healthcare professionals and the public has increased the importance of addressing public concern about its safety. Of particular concern has been the potential for transmission of infectious disease and organ and tissue injury, as well as the training and professional standards of acupuncture practitioners. This paper, therefore, addresses the following question: What is the frequency and severity of adverse complications and events in acupuncture treatment? DATA SOURCES: All first-hand case reports of complications and adverse effects of acupuncture that could be identified in the English language literature were reviewed and classified according to type of complication or adverse effect, circumstances of the event, credentials of the acupuncturist, country of occurrence, and long-term patient outcome. STUDY SELECTION: The case reports were selected by a search of 9 databases and covered the years between 1965-1999. DATA EXTRACTION: Relevant papers were collected and analyzed by 2 reviewers. Over the 35 years, 202 incidents were identified in 98 relevant papers reported from 22 countries. RESULTS: Types of complications included infections (primarily hepatitis from a few practitioners), and organ, tissue, and nerve injury. Adverse effects included cutaneous disorders, hypotension, fainting, and vomiting. There is a trend toward fewer reported serious complications after 1988. CONCLUSIONS: Declines in adverse reports may suggest that recent practices, such as clean needle techniques and more rigorous acupuncturist training requirements, have reduced the risks associated with the procedure. Therefore, acupuncture performed by trained practitioners using clean needle techniques is a generally safe procedure. PMID- 12564356 TI - Facility profile. Cancer center is patient friendly. PMID- 12564357 TI - The picture of health. PMID- 12564358 TI - Making the case. Selling maintenance and engineering initiatives to hospital executives. PMID- 12564359 TI - The future of the facility. Executive dialog series. PMID- 12564360 TI - Basic training. NFPA's fall education conference provides information for health facility professionals. PMID- 12564361 TI - Smooth move. Advice on easing the transition to team cleaning. PMID- 12564362 TI - Historical timeline 1853-2003. PMID- 12564363 TI - Frontier medicine. Texas doctors overcome disease and despair. PMID- 12564364 TI - 1953-2003. A tumultuous 50 years for medicine and TMA. PMID- 12564366 TI - UT tower shooting. A madman's rampage changes the emergency response system. PMID- 12564365 TI - Nov. 22, 1963. Texas physicians are thrust into history. PMID- 12564367 TI - Prairie to presidency. Women change the face of Texas medicine. PMID- 12564368 TI - Grouping together. Physicians move from solo to group practice. PMID- 12564369 TI - Committee traces history of Texas medicine. PMID- 12564370 TI - Into the unknown. Technology, continuing economic concerns color medicine's future. PMID- 12564371 TI - [A deadly pandemic is still spreading. The harmful effects of smoking are a serious warning for all]. AB - The author draw attention to the serious character of the pandemic caused by smoking which on a worldwide scale is responsible for the premature death of 4.2 million people every year. In the Czech Republic every year 22,000 people die from the sequelae of smoking. The authors refer to work which contributed to the evidence of the harmful impact of smoking on health. The World Health Organization contributes markedly to efforts to control smoking. The authors present brief information on the conference in Warsaw held in February 2002, they mention briefly data on smoking in Europe and the Czech Republic. They draw attention to the new strategy of the tobacco industry and appeal for joint and coordinated activities the objective of which would be to prevent health problems caused by smoking. PMID- 12564372 TI - [Present possibilities in vascular surgery]. AB - Contemporary vascular surgery relies on the progress in imaging techniques and on modern materials for vascular substitutes. The collaboration with radiodiagnosticians enabled the treatment of arterial thromboses by means of fibrinolysis. Radiodiagnostics plays an important role also in the field of endovascular operations, namely percutanous transluminal angioplasty, insertion of stents, and recently in the abdominal aorta aneurysms treatment by means of stents. Long-lasting effects of such operations, namely those in the aorta, have not been fully established and the classical reconstructive surgery has not lost its importance. PMID- 12564373 TI - [Intermittent androgen blockade in prostatic carcinoma]. AB - Intermittent endocrine treatment or cyclic therapy of prostate cancer aims at prolonging survival by delaying progression to androgen independence and at improving quality of life by avoiding the side effects of continuous androgen ablation. Intermittent endocrine treatment is capable of inducing multiple apoptoic regressions and of improvement in quality of life including sexual functions. The recommended length of therapeutic period is 6-9 months. The mean off-therapy interval approaches 50% of the duration of the treatment cycle. The mean time to disease progression is 32 month. The definitive answer to the question of prolonged survival awaits the completion of ongoing randomized studies. PMID- 12564374 TI - [Effects of nonsteroidal anti-inflammatory agents on the gastrointestinal tract]. AB - Non-steroidal anti-inflammatory drugs (NSAID) belong to the most commonly used drugs worldwide. NSAIDs can cause serious side effects to the gastrointestinal tract. During NSAIDs treatment 10-12% patients suffer from dyspepsia. Up to 1% patients develop severe gastrointestinal complications (ulcer, bleeding, perforation). Any part of gastrointestinal tract could be affected. In oesophagus, NSAIDs can cause oesophagitis of fibrous stricture. NSAID gastropathy can be detected in 40% patients chronically treated with NSAIDs. NSAIDs toxic injury to small and large bowel is frequent but only seldom properly recognised. Serious hepatic lesions are rare. There is no fully reliable and sure prophylaxis or treatment of NSAIDs impairment of to the gastrointestinal tract. Rate of side effects can be reduced by reasonable prescriptions and by primary and secondary prophylaxis. Low rate of side effects is associated with the use of pro-drugs (compound is metabolised to an active substance after absorption from the gastrointestinal tract). New promising drugs were developed with dual action (5 lipoxygenase- and COX-inhibition) and NSAID releasing NO (nitronaproxen, nitrophenac). Specific COX-2 inhibitors (coxibes) provide comparable anti inflammatory and analgesic effect but the risk of serious side effects to the gastrointestinal tract is significantly lower (when compared with non-specific NSAIDs). Beside harmful effects, NSAIDs are powerful tool in chemoprevention of colorectal cancer. PMID- 12564375 TI - [A phone survey of prostitute services in homes]. AB - BACKGROUND: Home prostitution is the hidden form of commercial sex network. We had done a phone survey of commercial sexual services provided in the private flats. METHODS AND RESULTS: We had used advertisements in the section "Intim services" of local adnewspaper "Avizo" as the primary source of relevant phone numbers. "Avizo" is sold in Northern Moravia--mainly in Ostrava, Karvina and Frydek-Mistek districts. We accomplished the phone interview with 48 subjects (individual persons or groups working in the particular flat) using the set of prepared questions. Three of these subjects were male, providing commercial sex for women. We estimated the number of such private flats, number of persons working there and number of professional contacts of this type in Karvina, Ostrava districts and whole Czech republic. CONCLUSIONS: We constructed an estimate of sexual sex workers for whole Czech republic--10,200 persons, therefrom 5100 persons in 1750 private flats, who accomplish to 1.36 mil sexual contact with customers annually. Regarding the risk behaviour nearly all of interviewed subjects indicated the use of condoms; 80% of subjects reported to be HIV-tested. PMID- 12564376 TI - [A statistical method for long-term monitoring of selected causes of death]. AB - The death rate statistics belongs to the essential health parameters and it is therefore frequently analysed. Authors suggest reviewing problems, which could be met, and discussing statistical methods in order to make the comparison and analysis of the development as much corresponding to the actual health state of the population as possible. Authors also discuss methods of the "International classification of diseases" and describe methods of direct data standardisation, including estimation of the variability. Using respiratory diseases as an example, authors illustrate the problems of revision of the code system in the "International classification of diseases" and the give examples of numerical evaluation. PMID- 12564377 TI - Manganese distribution across the blood-brain barrier. I. Evidence for carrier mediated influx of managanese citrate as well as manganese and manganese transferrin. AB - Manganese (Mn) is an essential element and a neurotoxicant. Regulation of Mn movement across the blood-brain barrier (BBB) contributes to whether the brain Mn concentration is functional or toxic. In plasma, Mn associates with water, small molecular weight ligands and proteins. Mn speciation may influence the kinetics of its movement through the BBB. In the present work, the brain influx rates of 54Mn2+, 54Mn citrate and 54Mn transferrin (54Mn Tf) were determined using the in situ brain perfusion technique. The influx rates were compared to their predicted diffusion rates, which were determined from their octanol/aqueous partitioning coefficients and molecular weights. The in situ brain perfusion fluid contained 54Mn2+, 54Mn citrate or 54Mn Tf and a vascular volume/extracellular space marker, 14C-sucrose, which did not appreciably cross the BBB during these short experiments (15-180 s). The influx transfer coefficient (Kin) was determined from four perfusion durations for each Mn species in nine brain regions and the lateral ventricular choroid plexus. The brain Kin was (5-13) x 10(-5), (3-51) x 10(-5), and (2-13) x 10(-5) ml/s/g for 54Mn2+, 54Mn citrate, and 54Mn Tf, respectively. Brain Kin values for any one of the three Mn species generally did not significantly differ among the nine brain regions and the choroid plexus. However, the brain Kin for Mn citrate was greater than Mn2+ and Mn Tf Kin values in a number of brain regions. When compared to calculated diffusion rates, brain Kin values suggest carrier-mediated brain influx of 54Mn2+, 54Mn citrate and 54Mn Tf. 55Mn citrate inhibited 54Mn citrate uptake, and 55Mn2+ inhibited 54Mn2+ uptake, supporting the conclusion of carrier-mediated brain Mn influx. The greater Kin values for Mn citrate than Mn2+ and its presence as a major non protein-bound Mn species in blood plasma suggest Mn citrate may be a major Mn species entering the brain. PMID- 12564378 TI - Manganese distribution across the blood-brain barrier. II. Manganese efflux from the brain does not appear to be carrier mediated. AB - There is concern about manganese (Mn) neurotoxicity. Mn can enter the brain by carrier-mediated influx. There have been no previous reports of investigation of Mn efflux from the brain. We used an established method that determines the rate of efflux out of the brain across the blood-brain barrier (BBB) from the product of the brain distribution volume (Vbrain) and the apparent elimination rate constant (Kel). Vbrain is determined as 54Mn uptake into rat parietal brain slices versus time. Kel is determined from the percentage of 54Mn remaining in the brain at various times after its discrete injection into the parietal cortex, compared to a reference compound which is expected to very slowly diffuse out of the brain. The Mn ion, Mn citrate and Mn transferrin (Mn Tf) were studied. 14C sucrose and 14C-dextran were used as reference compounds. The volume of distribution of the Mn species in brain slices was approximately 3-5 ml/g, indicating concentrative uptake. Mn, as the Mn ion or Mn citrate, was injected into the brain with sucrose or dextran to determine Kel. Based on the rapid exchange rate of Mn with ligands and on thermodynamic calculations, injection of Mn ion or Mn citrate into the brain would be expected to result in rapid formation of the same Mn species, predominantly the Mn ion, Mn citrates and Mn phosphate, in brain extracellular fluid. After injection into the brain Mn did not efflux from the brain more rapidly than sucrose or dextran, which diffuse across the BBB. Brain capillary diffusion of the Mn ion and Mn citrate would be expected to be slower than sucrose or dextran. The rate of Mn efflux from the brain is consistent with diffusion. PMID- 12564379 TI - EEG photic driving in workers exposed to mercury vapors. AB - OBJECTIVE: To assess the potential of EEG photic driving (PD) as an indicator of an early neurotoxic effect of long-term, low-level exposure to mercury vapors. SUBJECTS AND METHODS: Twenty-four chloralkali workers exposed to mercury vapors; twenty-four age- and gender-matched control subjects. Level of exposure was determined by urinary mercury excreted both spontaneously and after administration of a chelating agent, sodium 2,3-dimercapto-1-propane sulfonate. A computerized method for quantitative evaluation of PD was developed. Five parameters describing PD were compared. RESULTS: The number of stimulation frequencies eliciting PD was higher in the exposed group, with a median of 17 frequencies, as compared to 10 frequencies in the control group (P < 0.001). The maximum value of PD was higher in the exposed group, with a median of 24.6 z units as compared to 9.4 in the control group (P < 0.001). The median of the stimulation frequency with maximum PD was shifted from 15 Hz in the control group to 20 Hz in the exposed group (P < 0.01). The median of the sum of PD and the median of the index of PD were significantly higher in the exposed than in the control group (P < 0.001). The increased PD was particularly prominent at high stimulation frequencies in the beta range. There was no significant association between the measures of PD and the measures of exposure. CONCLUSIONS: In comparison with a control group, significantly increased photic driving was observed in a group of workers exposed to mercury vapors. The issue of whether or nor the intergroup differences in PD are mercury related, could not be determined on the basis of our results. Should the enhanced PD be caused by mercury, then this electrophysiological phenomenon might be regarded as a marker of the CNS hyperexcitability due to an early neurotoxic effect of mercury, the clinical expression of which is erethism. PMID- 12564380 TI - Effects of chronic lead exposure on the neuromuscular junction in Drosophila larvae. AB - Long term or chronic exposure to lead is associated with cognitive and other deficits in humans, which may reflect lead-induced changes in synaptic development and function. We believe that Drosophila has great potential as a model system for studying such changes. To test this, we compared the structure of single, identified synapses between identified axons (axons 1 and 2) and muscle fibers (fibers 6 and 7) in untreated 3rd instar larvae, and in larvae reared on medium made with 100 microM lead acetate in distilled water. We used three approaches to examine the motor terminals on muscle fibers 6 and 7 in segment 2: (1) all terminals were stained with an antibody to HRP; (2) only the terminals of axon 1 were stained by injecting biotinylated Lucifer yellow into it; and (3) the regions of the terminal containing synaptic vesicles were stained with an antibody to synaptotagmin, which provides an estimate of "synaptic" terminal area. Lead burdens were determined by inductively coupled plasma mass spectrometry; hemolymph lead levels at the neuromuscular junction were likely to be micromolar. We observed that lead exposure did not significantly affect the average terminal area or the average muscle fiber area, but did significantly affect the uniformity of the matching between muscle area and motor terminal size that normally occurs during development. There was a significant positive correlation between motor terminal size and muscle area in control, but not in lead-exposed larvae. The sensitivity of Drosophila larval synaptic development to lead opens the way to using the powerful genetic and molecular tools available for this system to study the underlying mechanisms of this sensitivity. We would hope that from such an understanding may come strategies for dealing with lead induced deficits in children. PMID- 12564381 TI - Muscle injury in organophosphorous poisoning and its role in the development of intermediate syndrome. AB - Muscle injury and its role in the development of Type II paralysis was studied in 25 patients with acute organophosphate poisoning. All patients were assessed for severity of poisoning at admission and through the course of poisoning for the development and duration of intermediate syndrome (IS) (Type II paralysis). Blood levels of acetylcholinesterase, creatine kinase, creatine kinase MM, LDH and LDH5 were estimated through the course of the poisoning. Of the 25 patients, 22 were severely poisoned and 3 had mild to moderate poisoning. Severely poisoned patients had a significantly greater rate of developing intermediate syndrome (17/22) (P = 0.026). Type I paralysis and fasciculations occurred in 76 and 70.5% of patients who developed intermediate syndrome, in comparison to 38 and 50%, respectively, of those who did not develop intermediate syndrome. Weakness developed in the same groups of muscles in both Types I and II paralysis but was of longer duration in patients who developed Type II paralysis. Acetylcholinesterase was inhibited > 90% throughout the course of poisoning with greater inhibition in patients with longer duration intermediate syndrome. Muscle injury was seen in all patients beginning at admission, peaking over the first 5 days and then declining over the next 5 days. Temporal profiles of blood muscle isoenzymes showed significantly greater muscle injury in those patients with greater severity of poisoning at admission, those who developed intermediate syndrome and in patients with longer duration intermediate syndrome. The findings of this study suggest that Types I and II paralysis in organophosphate poisoning are not separate syndromes but a clinical continuum determined by the severity of poisoning. The magnitude of organophosphate exposure and of muscle injury during the cholinergic crises appears to determine the occurrence and severity of intermediate syndrome. PMID- 12564382 TI - Effects of the new herbicide fentrazamide on the glucose utilization in neurons and erythrocytes in vitro. AB - Treatment of rats with fentrazamide for 2 years at 3000 ppm (males) and 4000 ppm (females) led to an increased incidence and degree of axonal degeneration in sciatic nerve as well as to effects on red blood cells. The mechanism underlying these effects was investigated in vitro using various cell cultures (permanent rodent cell lines from the nervous system, liver, kidney, skeletal and heart muscle and fibroblasts, primary cortical neurons and erythrocytes from the rat). Added to cultured rat cortical neurons for 1 week, fentrazamide considerably decreased glucose consumption, ATP levels and mitochondrial membrane potential and lowered the GSH level, however, it had little impact on viability and neurofilaments and did not induce oxidative stress (ROS) over the first 2 h. After recovery for 1 week, in addition some destruction of neurofilaments had occurred probably secondary to the disturbance of energy production. These effects were prevented by pyruvate. Further studies indicated that fentrazamide primarily inhibited glucose utilization, most likely by interfering with glycolysis. Similar effects were found in erythrocytes treated with fentrazamide over a period of 7 days. Primarily, the glucose consumption was reduced after 1 day treatment, followed by a marked reduction of the energy supply at days 3 and 7. Comparable to the neurons, the GSH level was significantly reduced. A marked hemolysis of the red blood cells was then observed after prolonged treatment. The extensive energy demand and exclusive dependency on glucose utilization of neurons and erythrocytes may explain the specific vulnerability of motor neurons and erythrocytes. When comparing the concentrations necessary for inducing effects in vitro on neuronal cells and erythrocytes to the very low plasma concentrations of fentrazamide in treated rats it is suggested that only a small impact of fentrazamide on the energy status at high doses will occur in vivo. Therefore, aging of the rat as another factor compromising mitochondrial energy production in motor neurons must be considered as additional contribution for the induction of axonal degeneration. It is concluded that this effect of fentrazamide in rats poses no specific risk under the exposure conditions relevant to humans. PMID- 12564383 TI - Estradiol enhances the neurotoxicity of glutamate in GT1-7 cells through an estrogen receptor-dependent mechanism. AB - Glutamate plays an important role in neuroendocrine regulation of reproduction through acting on the N-methyl-D-asparate receptor (NMDAR) in the preoptic area (POA). However, a larger dose of glutamate is neurotoxic. Estradiol (E2) increases the responsiveness of neurons to glutamate through activation and/or expression of NMDAR. In order to investigate whether estradiol modulates the neurotoxic effect of glutamate on the neurons through estrogen receptor (ER), immortalized GT1-7 cells, which simultaneously express ER and NMDAR were used. Tamoxifen and ICI 182,780, ER antagonist, were used to investigate whether the ER is involved in the effect of estradiol on glutamate-induced neurotoxicity. MK 801, a NMDAR antagonist, was used to confirm the enhancement of NMDAR-mediated neurotoxicity by estradiol. Neurotoxicity was evaluated by cell viability and LDH efflux. Cell death was observed by flow cytometry and DNA fragmentation. The results showed that: (1) estradiol (10 nM, incubated for 3 days) significantly enhanced the glutamate-induced neuronal death; (2) the percentages of necrosis and apoptosis were elevated after glutamate treatment, and estradiol significantly enhanced the glutamate-induced cell death; (3) glutamate-induced DNA fragmentation was enhanced by E2-pretreatment; (4) the induction of cell death and increase of LDH efflux after glutamate treatment were also enhanced by E2-pretreatment; (5) both the tamoxifen and ICI 182,780 abolished the estradiol enhanced NMDAR expression and neurotoxicity of glutamate; (6) higher dose of MK 801 (2 microM) was needed in E2-pretreated cells than in non-E2-pretreated group to block the glutamate-induced neurotoxicity. These results suggested that pretreatment of estradiol might enhance the expression of NMDAR and subsequent glutamate-induced neurotoxicity on the GT1-7 cells through an ER-dependent manner. PMID- 12564384 TI - Monoamine oxidase and cigarette smoking. AB - Current cigarette smokers have reduced monoamine oxidase (MAO) and there is evidence that this is a pharmacological effect of tobacco smoke exposure rather than a biological characteristic of smokers. This article summarizes human and animal studies documenting the inhibitory effects of tobacco smoke on MAO and discusses MAO inhibition in the context of smoking epidemiology, MAO inhibitor compounds in tobacco, reinvestigations of low platelet MAO in psychiatric disorders and smoking cessation. PMID- 12564385 TI - Voltage-dependent block of sodium channels in mammalian neurons by the oxadiazine insecticide indoxacarb and its metabolite DCJW. AB - Indoxacarb is a newly developed insecticide with high insecticidal activity and low toxicity to non-target organisms. Its metabolite, DCJW, is known to block compound action potentials in insect nerves and to inhibit sodium currents in cultured insect neurons. However, little is known about the effects of these compounds on the sodium channels of mammalian neurons. We compared the effects of indoxacarb and DCJW on tetrodotoxin-sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) sodium channels in rat dorsal root ganglion neurons by using the whole cell patch clamp technique. Indoxacarb and DCJW at 1-10 microM slowly and irreversibly blocked both TTX-S and TTX-R sodium channels in a voltage-dependent manner. The sodium channel activation kinetics were not significantly modified by 1 microM indoxacarb or 1 microM DCJW. The steady-state fast and slow inactivation curves were shifted in the hyperpolarization direction by 1 microM indoxacarb or 1 microM DCJW indicating a higher affinity of the inactivated sodium channels for these insecticides. These shifts resulted in an enhanced block at more depolarized potentials, thus explaining voltage-dependent block, and an apparent difference in the sensitivity of TTX-R and TTX-S channels to indoxacarb and DCJW near the resting potential. Indoxacarb and its metabolite DCJW cause toxicity through their action on the sodium channels. PMID- 12564386 TI - Butyl benzyl phthalate blocks Ca2+ signaling and catecholamine secretion coupled with nicotinic acetylcholine receptors in bovine adrenal chromaffin cells. AB - Butyl benzyl phthalate (BBP), a plasticizer and an environmental pollutant, exerts genomic estrogenic-like effects via estrogen receptors. In addition to exerting genomic effects via intracellular steroid receptors, estrogen exerts non genomic effects through interactions with membrane ion channels to lead the rapid alteration of neuronal excitability. Estradiol is known as to have modulating role on nicotinic acetylcholine receptors (nAChR). We investigated the possibility of BBP exerting non-genomic estrogenic-like effects on nAChR in bovine adrenal chromaffin cells. Our results show that BBP inhibited Ca2+ signaling induced by the nicotinic ligands carbachol, 1,1-dimethyl-4-phenyl piperazinium iodide (DMPP) and epibatidine (IC50 levels of 4.3, 4.1, 5.4 microM, respectively) as well as high K+ solution (IC50 50.9 microM). Additionally, in the electrophysiological observations, BBP blocked the inward current coupled with nAChR under the stimulation of carbachol. We, therefore, suggest that nAChR and voltage-gated Ca2+ channels are major and minor sites, respectively, of BBP action on the plasma membrane. The inhibitory effect of BBP on nAChR was found to be both noncompetitive and reversible, remaining unchanged as nAChR ligand concentration increased and decreased after washing. BBP was 10 times more potent than estradiol in inhibiting nAChR-coupled Ca2+ signals. We conclude that BBP exerts a novel rapidly inhibitory effect on nAChR. PMID- 12564387 TI - Acrylamide neuropathy. II. Spatiotemporal characteristics of nerve cell damage in brainstem and spinal cord. AB - Previous studies of acrylamide (ACR) neuropathy in rat PNS [Toxicol. Appl. Pharmacol. 151 (1998) 211] and cerebellum [NeuroToxicology 23 (2002) 397] have suggested that axon degeneration was not a primary effect and was, therefore, of unclear neurotoxicological significance. To continue morphological examination of ACR neurotoxicity in CNS, a cupric silver stain method was used to define spatiotemporal characteristics of nerve cell body, dendrite, axon and terminal degeneration in brainstem and spinal cord. Rats were exposed to ACR at a dose rate of either 50 mg/kg per day (i.p.) or 21 mg/kg per day (p.o.), and at selected times brains and spinal cord were removed and processed for silver staining. Results show that intoxication at the higher ACR dose-rate produced a nearly pure terminalopathy in brainstem and spinal cord regions, i.e. widespread nerve terminal degeneration and swelling were present in the absence of significant argyrophilic changes in neuronal cell bodies, dendrites or axons. Exposure to the lower ACR dose-rate caused initial nerve terminal argyrophilia in selected brainstem and spinal cord regions. As intoxication continued, axon degeneration developed in white matter of these CNS areas. At both dose-rates, argyrophilic changes in brainstem nerve terminals developed prior to the onset of significant gait abnormalities. In contrast, during exposure to the lower ACR dose-rate the appearance of axon degeneration in either brainstem or spinal cord was relatively delayed with respect to changes in gait. Thus, regardless of dose rate, ACR intoxication produced early, progressive nerve terminal degeneration. Axon degeneration occurred primarily during exposure to the lower ACR dose-rate and developed after the appearance of terminal degeneration and neurotoxicity. Spatiotemporal analysis suggested that degeneration began at the nerve terminal and then moved as a function of time in a somal direction along the corresponding axon. These data suggest that nerve terminals are a primary site of ACR action and that expression of axonopathy is restricted to subchronic dosing-rates. PMID- 12564388 TI - Acrylamide neuropathy. III. Spatiotemporal characteristics of nerve cell damage in forebrain. AB - Previous studies of acrylamide (ACR) neuropathy in rat PNS [Toxicol. Appl. Pharmacol. (1998) 151:211-221] and in spinal cord, brainstem and cerebellum [NeuroToxicology (2002a) 23:397-414; NeuroToxicology (2002b) 23:415-429] have suggested that axon degeneration was not a primary effect and was, therefore, of unclear neurotoxicological significance. To conclude our studies of neurodegeneration in rat CNS during ACR neurotoxicity, a cupric silver stain method was used to define spatiotemporal characteristics of nerve cell body, dendrite, axon and terminal argyrophilia in forebrain regions and nuclei. Rats were exposed to ACR at a dose-rate of either 50 mg/kg per day (i.p.) or 21 mg/kg per day (p.o.) and at selected times brains were removed and processed for silver staining. Results show that intoxication at either ACR dose-rate produced a terminalopathy, i.e. nerve terminal degeneration and swelling were present in the absence of significant argyrophilic changes in neuronal cell bodies, dendrites or axons. Exposure to the higher ACR dose-rate caused early onset (day 5), widespread nerve terminal degeneration in most of the major forebrain areas, e.g. cerebral cortex, thalamus, hypothalamus and basal ganglia. At the lower dose rate, nerve terminal degeneration in the forebrain developed early (day 7) but exhibited a relatively limited spatial distribution, i.e. anteroventral thalamic nucleus and the pars reticulata of the substantia nigra. Several hippocampal regions were affected at a later time point (day 28), i.e. CA1 field and subicular complex. At both dose-rates, argyrophilic changes in forebrain nerve terminals developed prior to the onset of significant gait abnormalities. Thus, in forebrain, ACR intoxication produced a pure terminalopathy that developed prior to the onset of significant neurological changes and progressed as a function of exposure. Neither dose-rate used in this study was associated with axon degeneration in any forebrain region. Our findings indicate that nerve terminals were selectively affected in forebrain areas and, therefore, might be primary sites of ACR action. PMID- 12564389 TI - The role of glycolysis and gluconeogenesis in the cytoprotection of neuroblastoma cells against 1-methyl 4-phenylpyridinium ion toxicity. AB - 1-Methyl-4-phenylpyridinium (MPP+) is a mitochondrial Complex I inhibitor and is frequently used to investigate the pathological degeneration of neurons associated with Parkinson's disease (PD). In vitro, extracellular concentration of glucose is one of the most critical factors in establishing the vulnerability of neurons to MPP+ toxicity. While glucose is the primary energy fuel for the brain, central nervous system (CNS) neurons can also take up and utilize other metabolic intermediates for energy. In this study, we compared various monosaccharides, disaccharides, nutritive/non-nutritive sugar alcohols, glycolytic and gluconeogenic metabolic intermediates for their cytoprotection against MPP+ in murine brain neuroblastoma cells. Several monosaccharides were effective against MMP+ (500 microM) including glucose, fructose and mannose, which restored cell viability to 109 +/- 5%, 70 +/- 5%, 99 +/- 3% of live controls, respectively. Slight protective effects were observed in the presence of 3-phosphoglyceric acid and glucose-6-phosphate; however, no protective effects were exhibited by galactose, sucrose, sorbitol, mannitol, glycerol or various gluconeogenic and ketogenic amino acids. On the other hand, fructose 1,6 bisphosphate and gluconeogenic energy intermediates [pyruvic acid, malic acid and phospho(enol)pyruvate (PEP)] were neuroprotective against MPP+. The gluconeogenic intermediates elevated intracellular levels of ATP and reduced propidium iodide (PI) nucleic acid staining to live controls, but did not alter the MPP(+)-induced loss of mitochondrial O2 consumption. These data indicate that malic acid, pyruvic acid and PEP contribute to anaerobic substrate level phosphorylation. The use of hydrazine sulfate to impede gluconeogenesis through PEP carboxykinase (PEPCK) inhibition heightened the protective effects of energy substrates possibly due to attenuated ATP demands from pyruvate carboxylase (PC) activity and pyruvate mitochondrial transport. It was concluded from these studies that several metabolic intermediates are effective in fueling anaerobic glycolysis during mitochondrial inhibition by MPP+. PMID- 12564390 TI - Asymmetrical development of the monoamine systems in 2,4-dichlorophenoxyacetic acid treated rats. AB - The purpose of this study was to determine whether the regional brain biogenic amine levels in adult rats were altered by pre- and post-natal exposure to 2,4 dichlorophenoxyacetic acid (2,4-D). Pregnant rats were daily orally exposed to 70 mg/kg per day of 2,4-D from gestation day (GD) 16 to post-partum day (PPD) 23. After weaning, the pups were assigned to one of two subgroups: T1 fed with untreated diet up to post-natal day (PND) 90 and T2 (maintained with 2,4-D diet up to PND 90). In addition, we wanted to know the effect of 2,4-D on lateralization in the monoamine systems of the basal ganglia of these adult rats and whether there was any correlation with the behavioral developmental pattern previously reported by us. In this study the content of noradrenaline (NA) was significantly increased in substantia nigra (SN) while it decreased in cerebellum in male and female rats of T2 group. The decreased dopamine (DA), 3,4 dihydroxyphenylacetic acid (DOPAC) and homovallinic acid (HVA) contents in cerebellum, midbrain, ventral tegmental area (VTA) and prefrontal cortex (PFc) showed an alteration in the mesocorticolimbic system. However, an increase of DA in SN and of DOPAC and HVA in nucleus accumbens (NAc) in both sexes and of DA and DOPAC (only in females) in striatum was detected. The contents of serotonin (5 hydroxytryptamine, 5-HT) were significantly increased in both sexes in PFc, striatum (St), midbrain, SN and cerebellum. Variations of any monoamine levels in NAc and VTA were determined. T1 rats were irreversibly altered: a diminution in DA and/or DOPAC levels in PFc, midbrain, VTA and cerebellum was determined. Indolamines of these rats were increased in both sexes in PFc and St. There was also a large increase in 5-HT levels in midbrain of male rats. Although no changes in the dopaminergic system with respect to their control values in any side of these brain structures were observed, DA and DOPAC levels were found to be decreased in the right side with respect to the left side in striata and accumbens nuclei in T2 female rats supporting the behavioral rotation previously registered by us in these rats. In addition, the increased 5-HT content detected in both the right and left striata observed in this study could be the answer to the behaviors observed and to the early alterations in dopamine in basal ganglia by 2,4-D in neonatal exposed rats, mediated by a serotonergic modulation on the dopaminergic system. PMID- 12564391 TI - Dante, qualitative research, and wonderings. PMID- 12564392 TI - The clergy as advocates for the severely demented. AB - The author sets forth the argument that pastoral caregivers ought to consider expanding their traditional role of ministering to dementia patients by pressing the issue of whether appropriate medication might significantly lessen the suffering of these persons. After discussing and documenting the current understanding regarding the nature of pain, the author outlines several advocacy, ethical, and procedural issues that could be included in providing pastoral care to this population, at the same time not miminizing more traditional faith-based activities. PMID- 12564393 TI - Nearly everything we wish our non-Jewish supervisors had known about us as Jewish supervisees. AB - The authors observe that Clinical Pastoral Education (CPE) developed out of a Protestant setting. Much of its thinking and writing therefore is heavily laden with Christian orientation and terminology. Sharing a general theological framework, most Christians read these words and think of the same- or similar ideas. However, Jews neither start with nor share the same theological beliefs. Jewish students perpetually ask themselves, "If the premise isn't true for me, can the conclusion still contain meaning?" Sometimes yes, sometimes no. Often, the resulting conflict leaves Jewish students feeling alienated from their CPE supervisors and peers. Few CPE supervisors realize that although everyone is reading the same material there are (at least) two "nations" present that are processing it differently. This article, by two National Association of Jewish Chaplains (NAJC) Board-Certified Rabbis, presents twelve key points about Judaism and Jewish thought to help non-Jewish CPE supervisors and chaplains in their work with Jewish supervisees and patients (residents, et al.). PMID- 12564394 TI - Pastoral hermeneutics and the challenge of a global economy: care to the living human Web. AB - The author discusses the relationship between a pastoral hermeneutics and the current social context as determined by international communication and globalization. He explores the influence of telecommunications on the human quest for meaning and the implication of this for pastoral care and counseling. A paradigm shift is proposed in terms of care to the living human web. A pastoral assessment which interprets the undergirding philosophy and belief system of globalization and its influence on human dignity is suggested; and a pastoral ministry which takes up its prophetic task and voices the needs of people in terms of a "globalization from below" is explicated. PMID- 12564395 TI - If chaplaincy understands health care reform, will it hurt the profession less? AB - The author summarizes the work of three authorities who present diverse views concerning health care reform. He suggests that these viewpoints will help chaplaincy more clearly understand clinical and administrative situations and promote creative adjustment to reform efforts. PMID- 12564396 TI - How Francoise Dolto links Lacanian psychoanalysis with the Christian Gospels. AB - This article introduces to an English-speaking audience of pastoral therapists, the writings of the French Lacanian psychoanalyst, Francoise Dolto (1908-1988) on the links she discovered between the most profound question raised by Lacanian psychoanalysis in its dynamics effects and the questions raised by the Christian Gospels. The author summarizes the main points of Dolto's Lacanian thought and where she departed from Lacan in her interpretation of the unconscious ethic of desire. Using Dolto's three writings on Lacanian psychoanalysis and the Bible, as well as material from her published clinical studies, the author illustrates Dolto's approach to the Bible, the parable of the Good Samaritan, and her application of the dialectical principles of desire in three case studies. PMID- 12564397 TI - Clinical Pastoral Education: reflections on the past and future of a movement. AB - In the year 2000, the celebration of seventy-five years of Clinical Pastoral Education (CPE) in the United States included attention to the history of the movement. A history of the past twenty-five years of the Association for Clinical Pastoral Education (ACPE) was published, plus several regional histories. This article grows out of the author's study of the history of the Northeast Region of ACPE in the context of the history of the Clinical Pastoral Education movement. The focus of this article is "the vision question"; that is, "What has happened to the visions of clinical training that were important in the beginning of the movement?" The history of the relations of clinical training to theological education is traced from the beginnings of CPE to the present time, with special emphasis on developments in the Northeast. Two implications of this historical review for the future of the movement are suggested. PMID- 12564398 TI - Mental health issues among clergy and other religious professionals: a review of research. AB - The authors reviewed the literature on mental health issues among clergy and other religious professionals, using electronic searches of databases of medical (Medline), nursing (CINAHL), psychology (PsycINFO), religious (ATLA), and sociological research (Sociofile). The existing research indicates the Protestant clergy report higher levels of occupational stress than Catholic priests, brothers, or sisters. Catholic sisters repeatedly reported the lowest work related stress, whereas women rabbis reported the highest stress levels in various studies. Occupational stress appears to be a source of family stress among Protestant clergy--a factor which clergy and their spouses believe the denominational leadership should address. High levels of stress also have been found to be associated with sexual misconduct among clergy. The authors make several recommendations based on these and other findings they report in their review. PMID- 12564399 TI - Drugs, motorcycles, God, & snow. PMID- 12564400 TI - She died with a tear in her eyes. PMID- 12564401 TI - Developing Medicare HMO market areas and their implications for HMO capitation rates. AB - Medicare+Choice (M+C) rates have replaced the average adjusted per-capita cost (AAPCC) payment method to HMOs as a result of the use of disparate benefit packages. However, the creation of M+C rates has likely caused HMOs to withdraw from the program. The underlying AAPCC problems are argued herein to be statistical in nature and arise because the AAPCC rates have been calculated on a county basis, although the county is generally not a health care market area. The author provides a method for empirically defining empirical market areas (EMAs), key principles to which one should adhere in developing these EMAs, and evidence that EMAs can serve as health care market areas. PMID- 12564402 TI - Electronic prescribing: implications for managed care. PMID- 12564403 TI - The combination of collaborative drug therapy management and E-prescribing. AB - Many discussions regarding electronic prescribing (E-prescribing) continue to focus on "who" should be responsible for building the communication platform: retail pharmacy or managed care. National pharmacy practice experts were interviewed to discuss a broader question: Given the potential for E-prescribing to reduce medication errors and improve communication between pharmacists and physicians, and given the increasing trend toward formal arrangements between pharmacists and physicians to establish collaborative drug therapy management (CDTM) practice settings, what are the implications for the practice of pharmacy? Specifically, how can technology for E-prescribing assist in the expansion of CDTM opportunities for the pharmacy profession? PMID- 12564404 TI - The role of independent community pharmacists in disease management. PMID- 12564405 TI - Recent tax changes may assist treatment of obesity. AB - The U.S. Surgeon General's Call to Action on Overweight and Obesity 2001 proposes inclusion of health care providers in prevention efforts and suggests classification of obesity as a disease category for reimbursement coding. Physician counseling on and referral for physical activity is generally recognized as a component of obesity treatment, but data on the effectiveness of these measures remain inconclusive. Recent changes in the federal tax law may influence physicians' and health plans' roles in promoting physical activity. PMID- 12564406 TI - A conversation with John B. Williams, MD, PhD. PMID- 12564407 TI - Aggressive treatment of type 2 diabetes: a managed care consensus statement. PMID- 12564408 TI - Burning fat through exercise. PMID- 12564409 TI - Taking charge of your emotions. PMID- 12564410 TI - All about toothpastes, toothbrushes, and mouthrinses. PMID- 12564411 TI - For parents. Parties and holidays. A map through the minefields. PMID- 12564412 TI - Supermarket smarts. Whole grains and whole grain products. PMID- 12564413 TI - Women & diabetes. Vaginitis. What every woman needs to know. PMID- 12564414 TI - Mast cell leukemia. PMID- 12564415 TI - [DCVG Congress in Bonn, September 11-l4, 2002. President's report: account and addenda]. PMID- 12564416 TI - [The 57th Congress of the DGVS, Bonn, September 14, 2002. Endoscopy Section Chairman's Report]. PMID- 12564417 TI - [Highlights of the 38th Annual Congress of the American Society of Clinical Oncology ASCO 2002]. PMID- 12564418 TI - Findings of scientific misconduct. PMID- 12564419 TI - Weekly oxaliplatin, high-dose folinic acid and 24h-5-fluorouracil (FUFOX) as salvage therapy in metastatic colorectal cancer patients pretreated with irinotecan and folinic acid/5-fluorouracil regimens. AB - Irinotecan (CPT-11), oxaliplatin (OXA) and different folinic acid(FA) modulated 5 fluorouracil (5-FU) regimens are active as first-and second-line chemotherapy of metastatic colorectal cancer. However, the best palliative sequence of these substances is still unclear. After CPT-11 containing regimens the optimal salvage protocol has not yet been defined. Here, we retrospectively analysed the weekly ambulant combination of OXA with continuous FA/5-FU (FUFOX) after two different CPT-11 containing chemotherapeutic regimens. PATIENTS: During October 1999 and May 2001, 20 patients (median 62; 48-74 years) were included who had disease progression after CPT-11 /bolus FA/5-FU (Saltz; 7 patients, group A) or after FA/5-FU followed by CPT-11 alone (13 patients, group B). OXA(60 mg/m2) was given for 2 hours prior to FA (500 mg/m2) as 2 h-infusion and continuous 5-FU (2.600 mg/m2) for 24 h-infusion on day 1,8,15 and 22 (repeated after week 6). RESULTS: FUFOX was administered 252 times. About 1,203 mg OXA per patient was given. Toxicities NC-CrC grade 3 were observed in 10 patients: diarrhoea (4), mucositis (5), nausea/vomiting (2), anaemia (1), leucopenia (1), thrombopenia (1) and hand foot-syndrome (1), 3 patients showed minor remissions, II patients stable disease. The median time to progression was 16(0-39) weeks. The median survival from start of FUFOX and from start of any palliative therapy was 33 (5-65) and 99 (44-200) weeks for all patients, respectively. CONCLUSIONS: FUFOX was efficient for additional tumour control in 70% of patients pretreated with CPT-11/5-FU based regimens. Sequential palliative treatment can lead to prolonged survival. PMID- 12564420 TI - [Imaging of the abdomen. AB - In the last decade, innovative ultrasound technology has become the primary imaging method in Gastroenterology. Compared to other imaging techniques (e. g., computed tomography and magnetic resonance imaging), conventional ultrasound images have the disadvantage of a restricted field-of-view (width), thus producing images that are not always easy to read. During the examination, the many mosaic-like single images obtained in real-time sonography must be mentally pieced together by the user, in order to obtain a complex 3-dimensional picture of the anatomical structure and its surrounding tissue. The extended field-of view of the Panoramic Imaging technique is a useful addition to conventional diagnostic ultrasound since it provides the enhanced overview required for exact topography of surrounding structures. This is especially true for the Power Mode Panoramic Imaging technique that allows the trading of vessels over greater distances, e.g., in the extremities. Thus, accuracy and reproducibility of measurements in the examination of larger structures and organs has improved. A clearer representation of these anatomical structures increases understanding and acceptance by referring clinicians. An exact diagnosis based on ultrasound imaging will certainly result in cost savings in health-care, since the use of further imaging methods (and therefore the total examination time) may be reduced. PMID- 12564421 TI - Differentiating diagnoses. PMID- 12564422 TI - [Catecholamine-dependent hereditary Cushing's syndrome - follow-up after unilateral adrenalectomy]. AB - CASE REPORT: We report on the laboratory, clinical, radiologic and histological findings of a 44-year-old male patient who was hospitalized with the clinical signs of Cushing's syndrome. The laboratory findings were suggestive of an adrenal genesis. Abdominal computed tomography revealed macronodular hyperplasia of both adrenal glands. Specific endocrinological tests demonstrated an increased secretion of cortisol during adrenergic stimulation which, however, was absent following beta-blocker medication. This would seem to point to an aberrant, beta receptor-mediated regulation of cortisol secretion as being the cause of increased cortisol production. ACTH-independent bilateral macronodular adrenocortical hyperplasia was also noted in the patient's mother. Neither the father nor the brother were affected by the disease. Apart from one case of familial appearance involving mother and daughter previously reported in the literature, this is the second description of a familial appearance and the first case involving mother and son. The patient underwent a left-sided adrenalectomy. Quick postoperative normalization of S-cortisol day profile and excretion of free dU-cortisol were noted under supportive temporary blockade of beta-receptors. In the further course, meanwhile spanning 2 1/4 years, a low basal cortisol secretion of the right adrenal gland has been observed, together with a lack of suppressibility in the dexamethasone suppression test. CONCLUSION: In the presence of mild clinical signs of adrenocortical insufficiency, matutinal administration of 5 mg hydrocortisone has proven sufficient for the past 6 months to reestablish a physiologic S-cortisol day profile with morning peak. PMID- 12564423 TI - To err is human. PMID- 12564425 TI - Rethinking coma cocktails. PMID- 12564426 TI - [Bleeding breast cancer - a rare cause of anemia in males]. PMID- 12564427 TI - [Europe's problems with scientific talent]. PMID- 12564428 TI - [Erythema gyratum repens. Drug hypersensitivity after azathioprine in a patient with type 1 autoimmune hepatitis]. PMID- 12564429 TI - Study is first to tie low health literacy with poor diabetes outcomes. PMID- 12564430 TI - Mammograms do save lives, says a new study. PMID- 12564431 TI - JCAHO issues call for action on nurse shortage and its effect on care quality. PMID- 12564432 TI - Docetaxel-related skin, nail, and vascular toxicity. PMID- 12564433 TI - Demographic factors and health coverage affect colorectal cancer treatment in the state of Washington. PMID- 12564434 TI - Publishing ethics: duplication indiscretion. PMID- 12564435 TI - [Multi-slice helical CT: image reconstruction algorithms, implementations, and clinical evaluations]. PMID- 12564436 TI - [Radiological quiz of the month]. PMID- 12564437 TI - [Phenotype variation of familial hypertrophic cardiomyopathy caused by combined missense mutations]. PMID- 12564438 TI - [Shortage of pediatricians]. PMID- 12564439 TI - Policy advice for Congress. PMID- 12564440 TI - One mother's perspective: Pattie Tobler. PMID- 12564441 TI - Is a market in human organs necessarily exploitative? PMID- 12564442 TI - Female genital mutilation and the moral status of abortion. PMID- 12564443 TI - Fairness and health care decision making: testing the group value model of procedural justice. AB - Considerable research has supported the notion that procedural justice is important to individuals independent of outcome considerations. Tyler and his associates (Lind and Tyler, 1988; Lind and Earley 1992; Tyler, 1989) proposed that this is so because procedural justice serves a group value function. In the present research, we explore aspects of this group value formulation in a real life health care context. Specifically, we assessed the relationship between individuals' appraisals of procedural justice following health care treatment decisions as well ass (i) expectations for different consequences to self; (ii) estimates of how they were viewed by the health care authorities; (iii) attributions about the reasons for their treatment; and (iv) ratings of emotional reactions. Results confirmed many components of the relational, group value model of procedural justice (Lind and Tyler, 1988; Lind and Earley, 1992; Tyler, 1989). Specifically, respondents who felt that they had been treated fairly expected that their status and their relationship with the health care decision maker and others in their health care group would improve; perceived that the health care decision maker would rate them positively on a variety of personality dimensions; felt that the health care decision maker's interactions with them revealed quite a lot about the decision-maker's personality characteristics; and indicated increased levels of pride and pleasure as well as lower levels of anger as a result of their treatment. Results were discussed from several frameworks including Markus and Kitayama's (1991) concept of independent and the interdependent self. PMID- 12564444 TI - The paradox of voluntary motherhood. PMID- 12564445 TI - Genetic manipulation in humans as a matter of Rawlsian justice. PMID- 12564446 TI - Rights of conscience in health care. PMID- 12564447 TI - Abortions and distortions: an analysis of morally irrelevant factors in Thomson's violinist thought experiment. PMID- 12564448 TI - [The enlightenment on pregnancy and birth in the 16th century]. AB - The enlightenment of people on pregnancy and childbirth is not just a contemporary phenomena. Henrik Smiths laegebog (Manual of Medicine) from 1557 is part of a tradition of communication and education going from antiquity to the present day. This book not only tells us the gynecological and obstetrical knowledge of a period, but is also an exciting reflexion of the perception of the same historical periods' view on gender, body and sexuality. PMID- 12564449 TI - [The human in the art: Henrik Have's painting for a medical thesis]. AB - A painting made by the Danish painter and author Henrik Have (born 1946) illustrates the front page of Jette E. Kristiansen's medical thesis The Antimicrobial Activity of Psychotherapeutic Drugs and Stereo-isomeric Analogues (1990). The painting illustrates beautifully that art and science can go hand in hand. Even very complicated chemical, pharmacological and microbiological questions can be expressed by means of colours and by means of symbols used in everyday life, such as a spiral (DNA), or a pair of hands expressing working together. Prayers, wishes and the most difficult questions in receptor stereo chemistry in eucaryotic and procaryotic cell-systems are illustrated in this painting. Synthetic chemistry and pharmacology are linked in the development of the synthetic dyes. The chemical colours are often the same in dyes and drugs. The red colour in the pharmacology is associated with the antibiotic drugs as sulfonamides as well as with the staining of Gram-negative bacteria. The yellow colour is associated with the antibiotic drugs, quinolones and the Ziehl-Neelsen staining for tubercle bacillii. The blue colour is associated with the psychoactive drugs, phenotiazines, as well as with methylenblue staining and the staining of Gram-positive bacteria. These association and symbols have been used in this painting. PMID- 12564450 TI - [Space perception: the history and its significance for art]. AB - Some aspects of the opthalmologic history of Denmark are briefly mentioned. Lens extraction in 1667 in Copenhagen, Edmund Hansen Gruts stereoophtalmoscopy in 1857, Ludvig Panums area in 1858 (single vision) space perception and outside this area double vision), squint treatment, Marius Tschernings periscopic spectacle lenses, Henning Ronnes stereoortograph and keiroscope and Gerhard Ronnes stereoscope. Space perception depends mostly on binocular function (convergens), but in fact some space perception occurs in vision due to perspective, accommodation, parallaxe, blurring, colours and shadows. The Danisk Poet, Hans Christian Andersen, has in his novels mentioned latent squint. The German poet Rainer Maria Rilke was much interested in perspective in connection with the development of impressionism, especially Paul Cezanne. Rilke in his later period developed the view that concentration on perspective removed those essential aspects from the world, in which he found God or a fourth dimension, as exemplified in the presence of ghosts. PMID- 12564451 TI - Bibliography. Current world literature. Leukemia. PMID- 12564452 TI - Bibliography. Current world literature. Cancer Biology. PMID- 12564453 TI - Bibliography. Current world literature. Endocrine tumors. PMID- 12564454 TI - It's a sin? Genetics and disabled people. PMID- 12564455 TI - Re: Gomez-Gerique, et al. Effect of atorvastatin and bezafibrate on plasma levels of C-reactive protein in combined (mixed) hyperlipidemia. Atherosclerosis 2002;162:245-51. PMID- 12564456 TI - Paradox posed. Psychiatric capacity shrinks as demand climbs. PMID- 12564457 TI - Clear call. FTC exempts not-for-profits from phone registry rule. PMID- 12564458 TI - Is it incorrect to report respiratory frequency in Hertz? PMID- 12564459 TI - Intracranial interventions. PMID- 12564460 TI - An Islamic view. PMID- 12564461 TI - A database of the innocent? PMID- 12564462 TI - Selling off the genome? PMID- 12564463 TI - Engineering consent. PMID- 12564464 TI - The new wave of colonialism. PMID- 12564465 TI - [Progress in the study of virtual impactor]. PMID- 12564466 TI - [Nutritional significance of biotransformation]. PMID- 12564467 TI - Volunteers of the Year. American Health Care Association. PMID- 12564468 TI - [Pierre Petit (1905-2002), pioneer of pediatric surgery and academic in spite of himself]. PMID- 12564469 TI - Synthesis of new hexahydro- and octahydropyrido[1,2-c]pyrimidine derivatives with an arylpiperazine moiety as ligands for 5-HT1A and 5-HT2A receptors. AB - Synthesis applied to prepare compounds 5-15 and 17-22 discussed in this paper has been presented in Scheme 1. Multi-stage preparation techniques were used to obtain 4-aryl-hexahydro 1-4 and (R,R) and (S,S) 4-aryl-octahydropyrido[1,2 c]pyrimidine-1,3-dione (16) derivatives, being the starting compounds for further modification. N-Alkylation of the imide group in compounds 1-4 and 16 followed, using 1,4-dibromobutane to yield monobromobutyl derivatives 5-8 and 17. Subsequent condensation of those compounds with appropriate 1-aryl or 1 heteroarylpiperazine led to the final hexahydro- 9-15 and octahydro- 18-22 pyrido[1,2-c]pyrimidine-1,3-dione derivatives. The final products were subjected to screening test to elucidate the affinity to 5-HT1A and 5-HT2A receptors. PMID- 12564470 TI - Synthesis antimicrobial and antifungal activity of some new 3 substituted derivatives of 4-(2,4-dichlorophenyl)-5-adamantyl-1H-1,2,4-triazole. AB - The synthesis of a series of substituted hydrazones and thiazolidinones is described, starting from N-[4-(2,4-dichlorophenyl)-5-adamantyl-1H-1,2,4-triazol-3 ylmercaptoacetyl]hydrazine. The new compounds were tested for antimicrobial and antifungal activity and some of them exhibited moderate activity against Candida albicans. PMID- 12564471 TI - Synthesis and binding affinities of 5-(3-pyridinyl)- and 5-(3-quinolinyl)-4 azahomoadamantanes to alpha7 nicotinic acetylcholine receptors. AB - A general synthetic route that can lead to nicotinic ligands bearing a variety of bulky aza-ring systems was developed. This methodology was applied to obtain 5-(3 pyridinyl)- and 5-(3-quinolinyl)-4-azahomoadamantanes 2a, 3a and 2b, 3b. The parent 5-(3-pyridinyl)-4-azahomoadamantane 2a (Ki = 5.0 microM) binds with about 100 times lower affinity than (+)-epibatidine 1 (Ki = 0.045 microM) to alpha7 nicotinic acetylcholine receptors (nAChRs). N-methyl substitution of 2a gives compound 3a which has about nine times lower binding affinity. The replacement of pyridinyl with a quinolinyl ring (compounds 2b, 3b) results in a dramatic reduction in potency (Ki > 1000 microM). PMID- 12564472 TI - Sustained release and biological availability of dalarelin from the biodegradable coacervate microcapsules. AB - A complex coacervation method was used to prepare microcapsules containing 74.8 +/- 1.5% of the 125I labelled dalarelin incorporated in the gelatine-algin coating. Microcapsules (62 +/- 1.7%) formed, did not exceed a size of 108 microm. The high content of the small size allowed this formulation to be administered by intramuscular injection to rats. It was found that the 125I labelled dalarelin in the form of microcapsules had better bioavailability and was active longer in the rat when compared with the 125I labelled dalarelin solution injections. Dalarelin administered in the microcapsular form was characterised by a higher biological availability. The degree of relative biological availability was calculated as 123% for the dalarelin in the microcapsular form. PMID- 12564473 TI - Alkyl diphenylacetyl, 9H-xanthene- and 9H-thioxanthene-carbonyl carbamates as positive allosteric modulators of mGlu1 receptors. AB - Starting from the random-screening hit 1a, a series of alkyl diphenylacetyl, 9H xanthene- and 9H-thioxanthene-carbonyl carbamates 1 has been prepared. These derivatives turned out to be selective positive allosteric modulators of mGlu1 receptors. These compounds do not directly activate mGlu1 receptors but markedly potentiate agonist stimulated responses, increasing potency and maximum efficacy. PMID- 12564474 TI - Synthesis of oxathiolane imidazole nucleosides. AB - Nucleosides have been of great interest since their strong antiviral activities were discovered. 1,3-Oxathiolane ring system has been known for many years, but it is in recent years that the ring has been used as the sugar ring in nucleoside analogs (Synthesis (1991) 1046; J. Am. Chem. Soc. 113 (1991) 9377; Tetrahedron Lett. 35 (1994) 4739). Besides, bredinin is a natural nucleoside antibiotic with imidazole moiety and there are some other studies reported on nucleosides with the imidazole group (Biorg. Med. Chem. 7 (1999) 481; Biorg. Med. Chem. 7 (1999) 1617; Nucleosides Nucleotides 18 (1999) 331). These findings make the imidazole group interesting as the base of a nucleoside. In this study, in order to find out the structure-activity relationships of L-oxathiolanyl nucleosides, L oxathiolanyl imidazole nucleosides 7 and 8 were synthesized, via novel intermediates 2-6, which were then tested for anti-HIV activity (Antivir. Res. 1 11 (1994) 25) in human peripheral blood mononuclear (PBM) cells, the synthesized nucleosides did not show significant activity up to 100 microM against HIV-1. PMID- 12564475 TI - Synthesis and pharmacological evaluation of novel antinociceptive N-substituted phenylimidazolyl-4-acylhydrazone derivatives. AB - This paper describes recent results of design, synthesis and pharmacological evaluation of new N-heterocyclic functionalized N-acylhydrazone compounds (NAH), belonging to the N-substituted-phenylimidazolyl-4-acylhydrazone class (3a-o). These compounds were planned by applying the molecular hybridization strategy to propose the structural modifications on the previously described functionalized 2 methyl-imidazolyl-3-acylhydrazone class (2), which presented an important analgesic profile. This new series (3) was synthesized in order to investigate the possible pharmacophoric contribution of the N-heteroaromatic ring and N acylhydrazone moieties to the analgesic activity. Compounds 3g and 3n are the most potent analgesic agents from this series, at the screening dose of 100 mg/kg p.o. and compounds 3e, 3j and 3o presented the best antiinflammatory properties at the same screening concentration. PMID- 12564476 TI - In vitro evaluation of thiazolyl and benzothiazolyl Schiff bases on pig cartilage. AB - A series of anti-inflammatory agents known as Schiff bases, combining thiazolyl and benzothiazolyl ring and vanillin moieties in the same molecule, was synthesized and evaluated for screening anti-degenerative activity on nasal pig cartilage cultures treated with interleukin 1beta, (IL-1beta). The amount of glycosaminoglycans (GAGs), the production of nitric oxide (NO) and prostaglandin E2 (PGE2), released into the culture medium, were detected. The tested Schiff bases decreased, dose-dependently, the NO and PGE2 production and the GAGs release with respect to samples treated with IL-1beta alone, showing a different behavior correlated to their structure. These results suggest that thiazolyl and benzothiazolyl Schiff bases in general, and particularly the Schiff base with bromine and methoxyl group in position three would protect cartilage matrix from degenerative factors induced by IL-1beta. PMID- 12564477 TI - Synthesis and antifungal properties of N-[(1,1'-biphenyl)-4-ylmethyl]-1H-imidazol 1-amine derivatives. AB - In the course of a study on 1H-imidazol-1-amine derivatives as antifungal agents, we found that N-[(1,1'-biphenyl)-4-ylmethyl]-N-[(2,4-dichlorophenyl)methyl]-1H imidazol-1-amine (1a) exhibited promising activities. In order to explore more in detail the structure-activity relationship of this new class of antifungal agents, we report now the synthesis and the biological activity of new analogues (1b-k) of compound 1a. The synthesis was performed using N-[(1,1'-biphenyl)-4 ylmethyl]-1H-imidazol-1-amine as starting material which was reacted with the proper arylmethyl halide. Most of the newly synthesized imidazolamines exhibited both fungal growth inhibition activity and cellular selectivity. PMID- 12564478 TI - Synthesis and biological activity of carbocyclic lexitropsins with a bioreductive fragment. AB - Carbocyclic oligopeptides containing of two, three or four aromatic rings with N,N-dimethylpropyl-1,3-diamine group as C-terminus fragment of compounds and 5 [bis(2-chloroethyl)amino]-2,4-dinitrobenzamide as N-terminal were synthesized. These lexitropsins present antitumour activity on the neoplastic cells hepatoblastoma HEP G2. These experiments were evaluated in hypoxic and oxygen conditions. Significant differences of activity in oxygen and hypoxic conditions were shown only in compound, N-(3-dimethylaminopropyl)-N'-([3-[5-bis(2 chlorethyl)amino]-2,4-dinitrobenzamide])-phenyl]urea dihydrochloride 1 (IC50 = 8545 nM in oxygen vs. IC50 = 710 nM in hypoxia). The rest of compounds (2-6) do not indicate differences of activity in oxygen and hypoxia. PMID- 12564479 TI - Infrared detection scheme with the photon avalanche effect. AB - We present a new infrared (IR) detection scheme based on the infrared quantum counter (IRQC) detector and utilizing the photon avalanche process. At the time of its discovery, the phenomenon of photon avalanche was considered a limitation rather than an advantage for the development of IRQC. Both the experimental results and the numerical modeling presented demonstrate that the process responsible for photon avalanche can be used to enhance the detection of an IR signal. A new room-temperature IR detection scheme is proposed on the basis of the results of this research. The novel detection scheme presented demonstrates an increase in detectivity and a decrease in the noise-equivalent power when compared with the IRQC schemes previously discussed in the literature. PMID- 12564480 TI - Analog tunable gratings driven by thin-film piezoelectric microelectromechanical actuators. AB - We present a microfabricated grating whose period can be tuned in analog fashion to within a fraction of a nanometer. The tunable angular range is more than 400 microrad in the first diffracted order. The design concept consists of a diffractive grating defined onto a 400-nm membrane, with the membrane subsequently strained in the direction perpendicular to the grating grooves by thin-film piezoelectric actuation. The strain-tuned grating device was fabricated with microelectromechanical processes, utilizing both surface and bulk micromachining. The fabricated piezoelectric film achieved a measured dielectric constant of 1200. Device characterization yielded grating period changes up to 8.3 nm (0.21% strain in the membrane) at 10 V and a diffracted angular change of 486 microrad, in good agreement with the theory. Uniformity across the actuated grating and out-of-plane deflections are characterized and discussed. PMID- 12564481 TI - Controlled modification and direct characterization of multimode-fiber refractive index profiles. AB - A combination of controlled annealing and characterization by scanning probe microscopy (SPM) is used to demonstrate that the refractive-index proffle of a commercially available silica-based optical fiber can be accurately reconfigured for use as an evanescent field sensor. The process relies on the controlled relocation of the silica glass dopants across the fiber cross section through heat treatment and the accurate measurement of the resulting dopant redistribution with SPM and differential etching techniques. The effect of variable annealing along a length of fiber is to produce a mode transformer to couple light from a laser source into the sensing region of the fiber. PMID- 12564482 TI - Chromatic dispersion and losses of microstructured optical fibers. AB - Using a rigorous and vector multipole method, we compute both losses and dispersion properties of microstructured optical fibers with finite cross sections. We restrict our study to triangular lattices of air-hole inclusions in a silica matrix, taking into account material dispersion. The fiber core is modeled by a missing inclusion. The influence of pitch, hole diameter, and number of hole rings on chromatic dispersion is described, and physical insights are given to explain the behavior observed. It is shown that flattened dispersion curves obtained for certain microstructured fiber configurations are unsuitable for applications because of the fibers' high losses and that they cannot be improved by a simple increase of the number of air-hole rings. PMID- 12564483 TI - High-speed path-length scanning with a multiple-pass cavity delay line. AB - Techniques for high-speed delay scanning are important for low-coherence interferometry, optical coherence tomography, pump probe measurements, and other applications. We demonstrate a novel scanning delay line using a multiple-pass cavity. Differential delays are accumulated with each pass so that millimeter delays can be generated with tens of micrometer mirror displacements. With special design criteria, misalignment sensitivity can be dramatically reduced. The system is demonstrated to scan 6 m/s at 2-kHz repetition rates. Real-time optical coherence tomography imaging with 500 pixel images at four frames/s is performed. Using a Cr:forsterite laser source, we obtained axial image resolutions of 6 microm with 92-dB sensitivity. PMID- 12564484 TI - Absorption of low-loss optical materials measured at 1064 nm by a position modulated collinear photothermal detection technique. AB - A collinear photothermal detection bench is described that makes use of a position-modulated heating source instead of the classic power-modulated source. This new modulation scheme increases by almost a factor 2 the sensitivity of a standard mirage bench. This bench is then used to measure the absorption coefficient of OH-free synthetic fused silica at 1064 nm in the parts per 10(6) range, which, combined with spectrophotometric measurements, confirms that the dominant absorption source is the OH content. PMID- 12564485 TI - Intercalibration of SUMER and CDS on SOHO. III. SUMER and CDS-GIS. Solar and Heliospheric Observatory. AB - Simultaneous observation of the same solar sources with different instruments is one way to test prelaunch radiometric calibrations and to detect changes in responsivity with time of extreme-ultraviolet instruments in space. Here we present the results of intercalibration of the SUMER (Solar Ultraviolet Measurements of Emitted Radiation) spectrometer (detectors A and B) and the GIS (Grazing Incidence Spectrometer), one of two spectrometers that compose the CDS (Coronal Diagnostic Spectrometer) on the Solar and Heliospheric Observatory (SOHO). The two instruments observed simultaneously radiances of emission lines at or near the center of the solar disk. The emission line chosen for intercomparison was Ne VIII at 770 A. However, such an intercomparison of the SUMER and CDS-GIS measurements means comparing two data sets with large differences in resolution and field of view. The latter difference, especially, introduces differences in the measured intensities caused by the solar variability that is relatively strong in the 770-A line. Using a statistical approach to overcome this problem, we found that the ratio of the GIS to the SUMER average radiances amounted to 2.6 +/- 0.9 before the SOHO's loss of attitude and to 2.1 +/- 0.7 afterward. These findings confirm earlier estimates of the GIS's responsivity being too low, and an update of the GIS calibration is recommended. Despite the large differences in resolution and field of view of the two instruments, the shapes of their normalized and rescaled histograms of the radiances agree well and therefore represent characteristic features of the Ne VIII line. PMID- 12564486 TI - Red-green-blue interferometer for the metrology of discontinuous structures. AB - Discontinuous surface profiles, e.g., diffractive optical elements (DOEs), are commonly measured by white-light interferometry. White-light interferometry needs significantly more memory capacity and computer time than does phase-shifting interferometry; there are approximately ten times more frames to be taken to gather the required information about the object under test. But usually the grooves of the DOEs are too deep for single-wavelength phase-shifting interferometry. Here we show how phase-shifting techniques can be applied to DOEs. For this purpose three interference patterns are recorded simultaneously by a three-chip color CCD camera at three wavelengths (Red-green-blue). It is possible to calculate separately the optical path difference at each pixel from the three phase patterns modulo 2pi. The algorithms used and experimental results are presented. PMID- 12564487 TI - Spectral interferometric microscope with tandem liquid-crystal Fabry-Perot interferometers for extension of the dynamic range in three-dimensional step height measurement. AB - The maximum measurable range of a spectral interference microscope depends on the coherence length of the light transmitted by its tunable spectral filter. To achieve a large range in step-height measurement we have developed a new tunable spectral filter that uses tandem liquid-crystal Fabry-Perot interferometers (LC FPIs), which can simultaneously attain both a high spectral resolution and a large tuning range. Fringe visibility measurements were carried out, and it was found that the coherence length of the light transmitted through tandem LC-FPIs is two times larger than that transmitted through a single LC-FPI. Using this novel tunable spectral filter, we developed a new spectral interference microscope for the measurement of three-dimensional shapes of discontinuous objects. Experimental results of step-height measurements both with a single LC FPI and with tandem LC-FPIs are presented for a combination of standard steel gauge block sets with 1-, 99-, and 100-microm steps. A large range (1-100 microm) of measurement with submicrometer resolution was achieved with tandem LC-FPIs that was not possible with our previous system in which a single LC-FPI was used. PMID- 12564488 TI - Comparison of test images obtained from various configurations of scanning near field optical microscopes. AB - The characteristics of a few experimental near-field optical microscopes, located in different laboratories, have been compared on the basis of their ability to image a well-defined submicrometer test object. PMID- 12564489 TI - Synthetic aperture interferometry: in-process measurement of aspheric optics. AB - A scanning probe consisting of a source and receive fiber pair is used to measure the phase difference between wave fronts scattered from the front and rear surfaces of an aspheric optic. This system can be thought of as a classical interferometer with an aperture synthesized from the data collected along the path of the probe. If the form of either surface is known, the other can be deduced. In contrast with classical interferometers, the method does not need test or null plates and has the potential to be integrated into the manufacturing process. PMID- 12564490 TI - Cryogenic optical testing of sandwich-type silicon carbide mirrors. AB - The experimental cryogenic performance of 160-mm-diameter silicon carbide (SiC) mirrors, one of which, a 700-mm-diameter mirror, is to be used as a primary mirror of the Japanese Infrared Astronomical Satellite ASTRO-F, is described. The mirrors are made from a sandwich-type SiC material that comprises a light porous core and a dense chemical-vapor-deposited coat of SiC. Three mirrors were manufactured consecutively, and changes in their surface contours related to temperature were measured with an interferometer when the mirrors were placed in a liquid-helium cryostat. Owing to significant improvements in manufacturing, the third SiC mirror showed only slight deformation as the temperature decreased from 300 to 6 K, which indicates high thermal strain homogeneity for a well-controlled sandwich-type SiC mirror. PMID- 12564491 TI - Ultrashort laser pulse beam shaping. AB - We calculated the temporal and spatial characteristics of an ultrashort laser pulse propagating through a diffractive beam-shaping system that converts a Gaussian beam into a beam with a uniform irradiance profile that was originally designed for continuous waves [Proc. SPIE 2863, 237(1996)]. The pulse front is found to be considerably curved for a 10-fs pulse, resulting in a temporal broadening of the pulse that increases with increasing radius. The spatial intensity distribution deviates significantly from a top-hat profile, whereas the fluence shows a homogeneous radial distribution. PMID- 12564492 TI - Characteristics of the thick, compound refractive lens. AB - A compound refractive lens (CRL), consisting of a series of N closely spaced lens elements each of which contributes a small fraction of the total focusing, can be used to focus x rays or neutrons. The thickness of a CRL can be comparable to its focal length, whereupon a thick-lens analysis must be performed. In contrast with the conventional optical lens, where the ray inside the lens follows a straight line, the ray inside the CRL is continually changing direction because of the multiple refracting surfaces. Thus the matrix representation for the thick CRL is quite different from that for the thick optical lens. Principal planes can be defined such that the thick-lens matrix can be converted to that of a thin lens. For a thick lens the focal length is greater than for a thin lens with the same lens curvature, but this lengthening effect is less for the CRL than for the conventional optical lens. PMID- 12564493 TI - Thermal forming of glass microsheets for x-ray telescope mirror segments. AB - We describe a technology to mass-produce ultrathin mirror substrates for x-ray telescopes of near Wolter-I geometry. Thermal glass forming is a low-cost method to produce high-throughput, spaceborne x-ray mirrors for the 0.1-200-keV energy band. These substrates can provide the collecting area envisioned for future x ray observatories. The glass microsheets are shaped into mirror segments at high temperature by use of a guiding mandrel, without polishing. We determine the physical properties and mechanisms that elucidate the formation process and that are crucial to improve surface quality. We develop a viscodynamic model for the glass strain as the forming proceeds to find the conditions for repeatability. Thermal forming preserves the x-ray reflectance and scattering properties of the raw glass. The imaging resolution is driven by a large wavelength figure. We discuss the sources of figure errors, and we calculate the relaxation time of surface ripples. PMID- 12564495 TI - Determination of glucose in whole blood samples by mid-infrared spectroscopy. AB - We have determined the glucose concentration of whole blood from mid-infrared spectra without sample preparation or use of chemical reagents. We selected 1119 1022 cm(-1) as the optimal wavelength range for our measurement by making a first loading vector analysis based on partial least-squares regression. We examined the influence of hemoglobin on samples by using different calibration and prediction sets. The accuracy of glucose prediction depended on the hemoglobin level in the calibration model; the sample set should represent the entire range of hemoglobin concentration. We obtained an accuracy of 5.9% in glucose prediction, and this value is well within a clinically acceptable range. PMID- 12564494 TI - SSR (spatially resolved refractometer): a null-seeking aberrometer. AB - The spatially resolved refractometer is an aberrometer used to measure the wave front aberrations of the human eye. In its original form and the new configuration that we report, it uses the patient's perception in a psychophysical task to evaluate the wave-front errors at a variable number of loci (typically 40 or 160) across the cornea. This configuration includes pupil tracking and the ability to choose the measurement loci in software. An automated configuration that does not require patient input is also described. PMID- 12564496 TI - Optical imaging based on time-resolved Stokes vectors in filamentous tissues. AB - Time-resolved Stokes vector components of light transmitted through filamentous tissues were measured with a view to improving the imaging quality of optical images in such tissues. Temporal profiles of the Stokes vectors and the time resolved degree of polarization (DOP) were calibrated to produce higher image quality than that of images based on time gating, polarization discrimination, or both. A thin chicken bone inserted into chicken breast tissue with filament orientation in different directions with respect to the direction of input linear polarization was scanned to demonstrate images of higher spatial resolution and contrast based on the measurement of time-resolved DOP. PMID- 12564497 TI - Optimization of a beam shaping bolus for superficial microwave hyperthermia waveguide applicators using a finite element method. AB - Temperature inhomogeneity in hyperthermia treatments often limits the total thermal dose that can be delivered to the tumour region. To reduce such inhomogeneities, a prototype dynamically modifiable square array of saline-filled patches which attenuate microwave energy was developed for superficial treatments that use external microwave applicators. The array was situated inside the coupling water bolus that is often used with external applicators. The prototype has been previously tested clinically with promising results. A more complete theoretical analysis of the performance of this new bolus design and improvements to its design by modelling are presented here. The analysis was performed by performing five iterative simulations of the SAR pattern produced inside a tissue structure by a waveguide applicator with a water bolus containing the dynamic patch array attached. Between iterations the patch array configuration was modified in an attempt to improve the ability of the bolus to confine heating to an 'L'-shaped tumour region. These simulations were performed using the finite element method. The steady-state temperature profile was then computed using a finite element method based simulation of heat transfer that assumed a given applicator power level and water bolus temperature. Several iterations of these heat transfer simulations were performed with varying applicator power level and water bolus temperature to improve the confinement of heating to the target region. The analysis showed that the dynamic patch array should be capable of conforming heating to an 'L'-shaped target tumour region while limiting the heating to the surrounding normal tissue to an acceptable level. PMID- 12564498 TI - Rate process analysis of thermal damage in cartilage. AB - Cartilage laser thermoforming (CLT) is a new surgical procedure that allows in situ treatment of deformities in the head and neck with less morbidity than traditional approaches. While some animal and human studies have shown promising results, the clinical feasibility of CLT depends on preservation of chondrocyte viability, which has not been extensively studied. The present paper characterizes cellular damage due to heat in rabbit nasal cartilage. Damage was modelled as a first orderrate process for which two experimentally derived coefficients, A = 1.2 x 10(70) s(-1) and Ea = 4.5 x 10(5) J mole(-1), were determined by quantifying the decrease in concentration of healthy chondrocytes in tissue samples as a function of exposure time to constant-temperature water baths. After immersion, chondrocytes were enzymatically isolated from the matrix and stained with a two-component fluorescent dye. The dye binds nuclear DNA differentially depending upon chondrocyte viability. A flow cytometer was used to detect differential cell fluorescence to determine the percentage of live and dead cells in each sample. As a result, a damage kinetic model was obtained that can be used to predict the onset, extent and severity of cellular injury to thermal exposure. PMID- 12564499 TI - MRI feedback temperature control for focused ultrasound surgery. AB - A temperature feedback controller routine using a physical model for temperature evolution was developed for use with focused ultrasound surgery. The algorithm for the controller was a multi-input, single-output linear quadratic regulator (LQR) derived from Pennes' bioheat transfer equation. The controller was tested with simulated temperature data that had the same characteristics as those obtained with magnetic resonance imaging (MRI). The output of the controller was the appropriate power level to be used by the transducer. Tissue parameters estimated prior to the simulated treatments were used to determine the controller parameters. The controller performance was simulated in three dimensions with varying system parameters, and sufficient temperature tracking was achieved. The worst-case overshoot was 7 degrees C and the steady-state error was 5 degrees C. The simulated behaviour of the controller suggests satisfactory performance and that the controller may be useful in controlling the power output during MRI monitored ultrasound surgery. PMID- 12564500 TI - Acquiring a four-dimensional computed tomography dataset using an external respiratory signal. AB - Four-dimensional (4D) methods strive to achieve highly conformal radiotherapy, particularly for lung and breast tumours, in the presence of respiratory-induced motion of tumours and normal tissues. Four-dimensional radiotherapy accounts for respiratory motion during imaging, planning and radiation delivery, and requires a 4D CT image in which the internal anatomy motion as a function of the respiratory cycle can be quantified. The aims of our research were (a) to develop a method to acquire 4D CT images from a spiral CT scan using an external respiratory signal and (b) to examine the potential utility of 4D CT imaging. A commercially available respiratory motion monitoring system provided an 'external' tracking signal of the patient's breathing. Simultaneous recording of a TTL 'X-Ray ON' signal from the CT scanner indicated the start time of CT image acquisition, thus facilitating time stamping of all subsequent images. An over sampled spiral CT scan was acquired using a pitch of 0.5 and scanner rotation time of 1.5 s. Each image from such a scan was sorted into an image bin that corresponded with the phase of the respiratory cycle in which the image was acquired. The complete set of such image bins accumulated over a respiratory cycle constitutes a 4D CT dataset. Four-dimensional CT datasets of a mechanical oscillator phantom and a patient undergoing lung radiotherapy were acquired. Motion artefacts were significantly reduced in the images in the 4D CT dataset compared to the three-dimensional (3D) images, for which respiratory motion was not accounted. Accounting for respiratory motion using 4D CT imaging is feasible and yields images with less distortion than 3D images. 4D images also contain respiratory motion information not available in a 3D CT image. PMID- 12564501 TI - Refined gypsum effective attenuation coefficients for Co-60, 6 MV, 10 MV and 18 MV x-rays. AB - We report new effective linear attenuation coefficients (mueff) for refined gypsum for Co-60 and 18 MV x-rays. 6 MV and 10 MV mueff agree with published data. For a 100 cm2 field size (FS), tissue defect x = 1.35 cm, target depth d = 1.65 cm beneath the tissue defect x, mueff is 0.123 cm(-1) (Co-60) and 0.0934 cm( 1) (6 MV). For 100 cm2, x = 1.35 cm, and d = 5.65 cm beneath x, mueff is 0.072 cm(-1) (10 MV), and 0.0614 cm(-1) (18 MV). Ford, mueff decreases about 10% from 25 to 400 cm2. For a given FS, mueff decreases with d by 3%-5% for Co-60, and 3% for 6 MV, 10 MV and 18 MV, but depends on (d - x). For d, when x is large (8 cm), depending on energy and FS, mueff is 2%-4% less than when x is small (2 cm). These data were used in a treatment-planning computer to design compensator filters for a step phantom. Compensation was to within 10% in the compensation plane (CP). Above and below CP, computer-calculated ratios of doses with and without filters were 0.75-1.13. Chamber dose ratios with and without filters were 0.75-1.12. PMID- 12564502 TI - Integral-transport-based deterministic brachytherapy dose calculations. AB - We developed a transport-equation-based deterministic algorithm for computing three-dimensional brachytherapy dose distributions. The deterministic algorithm has been based on the integral transport equation. The algorithm provided us with the capability of computing dose distributions for multiple isotropic point and/or volumetric sources in a homogenous/heterogeneous medium. The algorithm results have been benchmarked against the results from the literature and MCNP results for isotropic point sources and volumetric sources. PMID- 12564503 TI - Effect of regional differences in cardiac cellular electrophysiology on the stability of ventricular arrhythmias: a computational study. AB - Re-entry is an important mechanism of cardiac arrhythmias. During re-entry a wave of electrical activation repeatedly propagates into recovered tissue, rotating around a rod-like filament. Breakdown of a single re-entrant wave into multiple waves is believed to underlie the transition from ventricular tachycardia to ventricular fibrillation. Several mechanisms of breakup have been identified including the effect of anisotropic conduction in the ventricular wall. Cells in the inner and outer layers of the ventricular wall have different action potential durations (APD), and support re-entrant waves with different periods. The aim of this study was to use a computational approach to study twisting and breakdown in a transmural re-entrant wave spanning these regions, and examine the relative role of this effect and anisotropic conduction. We used a simplified model of action potential conduction in the ventricular wall that we modified so that it supported stable re-entry in an anisotropic model with uniform APD. We first examined the effect of regional differences on breakdown in an isotropic model with transmural differences in APD, and found that twisting of the re entrant filament resulted in buckling and breakdown during the second cycle of re entry. We found that breakdown was amplified in the anisotropic model, resulting in complex activation in the region of longest APD. This study shows that regional differences in cardiac electrophysiology are a potentially important mechanism for destabilizing re-entry and may act synergistically with other mechanisms to mediate the transition from ventricular tachycardia to ventricular fibrillation. PMID- 12564504 TI - A novel method of studying total body water content using a resonant cavity: experiments and numerical simulation. AB - A novel electromagnetic method of obtaining total body water is proposed, in which the water content is obtained from the dielectric properties as measured by a resonant perturbation technique. A screened room acts as a radio-frequency cavity, in our case resonating at 59 MHz, a frequency at which both real and imaginary parts of the complex permittivity of tissues are correlated to their moisture content. The presence of a human subject in the room leads to both a negative shift in the room's resonant frequency and a reduction in its Q-factor. We simulated the room and the body using the transmission line matrix (TLM) method, a computational electromagnetic code which models the problem in the time domain. Experiment and numerical model showed good agreement for two orientations of the subject. The sensitivity of the technique was investigated by measuring the response before and after the subject drank a small quantity of water, less than 2% of body mass. The resulting change in the resonant frequency was significant, and was also predicted by the numerical model. The proposed technique for studying body composition is simple, non-invasive and employs non ionizing radio waves at low power. PMID- 12564505 TI - Estimation of the supplementary axial wall stress generated at peak flow by an arterial stenosis. AB - Mechanical stresses in arterial walls are known to be implicated in the development of atherosclerosis. While shear stress and circumferential stress have received a lot of attention, axial stress has not. Yet, stenoses can be intuitively expected to produce a supplementary axial stress during flow systole in the region immediately proximal to the constriction cone. In this paper, a model for the estimation of this effect is presented, and ten numerical examples are computed. These examples show that the cyclic increase in axial stress can be quite considerable in severe stenoses (typically 120% or more of the normal stress value). This result is in best agreement with the known mechanical or morphological risk factors of stenosis progression and restenosis (hypertension, elevated pulse pressure, degree of stenosis, stenosis geometry, residual stenosis, etc). The supplementary axial stress generated by a stenosis might create the damages in the endothelium and in the elastic membranes which potentiate the action of the other risk factors (hyperlipidaemia, diabetes, etc). It could thus be an important cause of stenosis progression and of restenosis. PMID- 12564506 TI - Effects of mass flow rate and droplet velocity on surface heat flux during cryogen spray cooling. AB - Cryogen spray cooling (CSC) is used to protect the epidermis during dermatologic laser surgery. To date, the relative influence of the fundamental spray parameters on surface cooling remains incompletely understood. This study explores the effects of mass flow rate and average droplet velocity on the surface heat flux during CSC. It is shown that the effect of mass flow rate on the surface heat flux is much more important compared to that of droplet velocity. However, for fully atomized sprays with small flow rates, droplet velocity can make a substantial difference in the surface heat flux. PMID- 12564507 TI - Comparison of dose rates calculated on Nucletron NPS v11: catheter tracking versus catheter describing. PMID- 12564508 TI - Comment on 'Comparison of dose rates calculated on Nucletron NPS v11: catheter tracking versus catheter describing'. PMID- 12564509 TI - A vision for tinnitus research. PMID- 12564510 TI - Psychiatric comorbidity in a population of outpatients affected by tinnitus. AB - The aim of this study was to outline the psychopathological characteristics of a population of outpatients affected by tinnitus and to consider its impact on their mental state and ability to function in major areas of their lives. Seventy five consecutive tinnitus patients were enrolled on their first visit to the outpatients clinic of the Audiology Department of the 'Federico II' University of Naples, for audiological and psychiatric evaluation. A series of audiometric and vestibular tests was performed for tinnitus rating assessment, and further information was obtained from the patient via a semi-structural interview. For the psychopathological examination, patients underwent the Mini International Neuropsychiatric Interview (MINI), by means of which a multiaxial diagnosis (five axes) was expressed, according to the criteria of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). For a better understanding of the personality aspects, the Minnesota Multiphasic Personality Inventory (MMPI) test was administered to a subgroup of 55 subjects whose cultural background permitted their full cooperation. The results of the study show that 58 subjects (77% of the total) met the criteria for psychiatric disorder diagnosis, according to the DSM-IV system: Axis I comprises anxiety, affective and somatoform disorders and psychoses; Axis II comprises personality disorders. Multiple diagnoses were expressed in some subjects. The findings of the examination via MMPI show a high percentage of depression, hysteria, and hypochondria. Although we are not seeking to establish a cause-effect relationship between the unpleasant experience of tinnitus and psychopathological disorders, our findings are consistent with those of other authors. Tinnitus can indeed have severe consequences for the subject's ability to function in many areas of their life. In this paper, the implications of such results for the diagnosis and therapy of tinnitus are discussed. PMID- 12564511 TI - Design, optimization and evaluation of a Danish sentence test in noise. AB - The Danish sentence test DANTALE II was developed in analogy to the Swedish sentence test by Hagerman and the German Oldenburg sentence test as a new Danish sentence test in noise to determine the speech reception threshold in noise (SRT, i.e. the signal-to-noise ratio (SNR) that yields 500% intelligibility). Each sentence is generated by a random combination of the alternatives of a base list. This base list consists of 10 sentences with the same syntactical structure (name, verb, numeral, adjective, object). The test sentences were recorded and segmented in such a way that the coarticulation effects were taken into account in order to achieve a high perceived sound quality of the resynthesized sentences: 100 sentences were recorded, each coarticulation between each word and the 10 possible following word alternatives were recorded, and the correct coarticulation was used to generate the test sentences. Word-specific speech recognition curves were measured for each recorded word to optimize the homogeneity of the speech material and the measurement accuracy. Level corrections of particular words and a careful selection of the test lists produced a noticeable reduction in the variation in the distribution of word specific SRT (standard deviation 1.75 dB instead of 3.78 dB). Therefore, the slope of the total intelligibility function was expected to increase from 8.30%/dB (raw test material) to 13.2%/dB (after modification). These theoretical expectations were evaluated by independent measurements with normal-hearing subjects, and, for the most part, confirmed. The reference data for the DANTALE II are: SRT=-8.43 dB SNR; slope at SRT, s50 = 13.2%/dB. The training effect was 2.2 dB and could be reduced to less than 1 dB if two training lists of 20 sentences were performed prior to data collection. PMID- 12564512 TI - Prevalence of hearing impairment in a population in Sweden. AB - An epidemiological study was carried out, based on 590 randomly selected subjects, aged from 20 to 80 years, in the province of Ostergotland in Sweden. The results obtained were similar to published results from other countries, with an overall prevalence of subjects with average hearing threshold levels over the frequencies 0.5, 1, 2 and 4 kHz (M4) equal to or exceeding 25 dB HL for the better ear (BE) of 16.9% (95% CI: 13.819.9%). For M4 BE > or = 35 dB HL, the prevalence was 7.7% (95% CI: 5.5-9.8%), for M4 BE > or = 45 dB HL, the prevalence was 3.3% (95% CI: 1.9-4.8%), and for M4 BE > or = 65 dB HL, the prevalence was 0.2% (95% Cl: 0.0-0.6%). The overall prevalence of reported tinnitus was 13.2% (95% CI: 10.5-16.0%). In the population under study, 7.7% of the subjects were estimated to benefit from a hearing aid, while the prevalence of hearing aid users was 2.4%. PMID- 12564513 TI - Temporary threshold shift in military pilots measured using conventional and extended high-frequency audiometry after one flight. AB - Noise of such a high level that it can result in hearing deterioration is an inherent characteristic of military flying. Susceptibility to hearing impairment was studied using 51 Finnish Air Force military personnel as subjects. The test persons flew missions on a British Aerospace Hawk 51 advanced jet trainer, Boeing F-18 Hornet, Mikoyan & Gurevich MiG-21bis and Saab 35 Draken interceptors, and a Valmet Redigo turboprop liaison aircraft. The duration of noise exposure was one flight mission, which varied from 30 to 60 min. Noise doses and levels were measured using a miniature microphone at the inlet of the ear canal, while a second microphone was located at the level of the subject's shoulder. Hearing thresholds were measured before each flight using conventional (0.125-8 kHz) and extended high-frequency (EHF) (8.20 kHz) audiometry. The measurements were repeated as soon as possible after the flight. The study showed that the pre flight threshold levels of the subjects were good. Both conventional and EHF audiometry revealed statistically significant temporary threshold shifts (TTS) at several frequencies and with all aircraft types involved. The changes were, however, minor. The risk of noise-induced hearing impairment at the studied exposure levels is, in all probability, rather small. The role of extended high frequency audiometry would be in research, and it might be performed for flying personnel upon entering service and every fifth year thereafter. PMID- 12564514 TI - Evaluation of the noise reduction system in a commercial digital hearing aid. AB - We evaluated the effectiveness of a noise reduction system implemented in a commercial digital multichannel compression hearing aid. Eight experienced hearing aid wearers with moderate sensorineural hearing loss were fitted bilaterally according to the manufacturer's fitting guidelines. After a 3-month period of regular use of two programs, one with and one without the noise reduction system, speech recognition thresholds (SRTs) were measured in four types of background noise, including steady noise, and noises with spectral and/or temporal dips. SRTs were very similar with and without the noise reduction system; in both cases, SRTs were markedly lower than for unaided listening. SRTs were lower for the noises with dips than for the steady noise, especially for the aided conditions, indicating that amplification can help to 'listen in the dips'. Ratings of sound quality and listening comfort in the aided conditions were uniformly high and very similar with and without the noise reduction system. PMID- 12564515 TI - Analysis of programming maps and its application for balancing multichannel cochlear implants. AB - Channel balancing is important for correct perception when using multichannel cochlear implants. In this paper we analyse the effect of channel imbalance on hearing perception in cochlear-implanted patients. Based on warble tone audiometry measured for 10 implanted patients, we evaluate how hearing sensitivity is affected by an imbalance. We also study the distribution along the cochlea of the programming parameters for 146 patients. The parameters are compared for different groups of patients (defined taking into account the age at implantation, the stimulation mode, and the presence of severe cochlear damage). The aim of this study is to obtain information that can be useful for balancing the channels for the patients in the different groups, as well as to evaluate the effect of an imbalance on perception. These results provide some complementary information that can be applied to the correct fitting of the cochlear implant processor. PMID- 12564516 TI - Misconceptions when estimating the directivity index for directional microphone systems on a manikin. PMID- 12564517 TI - Reflections on life as a District Health Board member. PMID- 12564518 TI - Meeting the challenges of critical case study in nursing research: an interview with Alison Dixon. AB - Critical case study is a methodology seldom used in nursing or midwifery research, yet it offers a process for revealing and acting on power relations in the practice worlds of nurses and midwives. This is the eighth article in a series based on interviews with nursing and midwifery researchers, designed to provide the beginning researcher with a first-hand account of the experience of using particular methodologies. This article focuses on critical case study as experienced by Alison Dixon (RGON, BA, Dip Soc Sci (Nursing), PhD) who used this methodology to explore the difference in practice between enrolled and registered nurse practice. PMID- 12564519 TI - Measuring perceptions of the clinical career pathway in a New Zealand hospital. AB - Clinical Career Pathways (CCPs) for nurses were introduced in the 1970s and they were first established in New Zealand during the late 1980s. The implementation of CCP programmes has met with mixed response; many nurses view it negatively as an extra and unnecessary demand from their employers while others perceive it to be a valuable form of professional development. This paper introduces a new instrument, the Clinical Career Pathway Evaluation Tool (CCPET) designed to assess nurses' and midwives' knowledge of and attitudes towards their Clinical Career Pathway. The 51 item instrument takes the form of a self-report questionnaire in two sections. The first tests knowledge of the CCP, as implemented at the study hospital, the second measures attitudes towards CCP and professional development. In this paper we describe the development of the CCPET and present some of the results from an initial application of the instrument with 239 nurses and midwives in a New Zealand hospital. Results indicate that knowledge levels were moderate in this sample and were correlated with both positive and negative attitudes. Results of t-test comparisons indicated that, on average, the group who had already completed a CCP portfolio had greater knowledge and more positive attitudes than the group who had not. The authors suggest firstly that the CCPET is useful for measuring CCP knowledge and attitudes in a constantly restructuring nursing environment, and secondly that the instrument can be easily adapted for use in other hospitals and organisations. PMID- 12564520 TI - Defining currency of practice for nurse educators. AB - Recent Nursing Council of New Zealand (NCNZ) guidelines for competence-based practising certificates (NCNZ, 2001) and the fact that all nurse educators must have a current practising certificate prompted the Nursing Schools within the Tertiary Accord of New Zealand (TANZ) to explore issues surrounding current competency in practice and how this can be maintained by nurse educators. This is a topical debate as discussions related to competence-based practising certificates generally refer to competence only in terms of direct patient care. This article sets out to clarify the issue with specific reference to nurse educators who, by the nature of their scope of practice, often do not carry a patient caseload. The literature relating to currency of practice is explored to provide background to current definitions and existing strategies for maintaining competence. This article draws on the findings of a survey by the TANZ Nursing Schools and provides a position on how currency of practice applies to nurses working in educational settings. In addition, strategies to maintain clinical, teaching and scholarly currency are presented along with suggestions for providing evidence that currency of practice is maintained. PMID- 12564521 TI - South Auckland community paediatric nursing service: a framework for evaluation. AB - This paper describes the Kidz First paediatric community homecare nursing team in South Auckland. While the service was not initially planned as an integrated approach to child health, its evolution reflects the move to more community based care delivery and the expansion of nurse-led initiatives in New Zealand. The components of a community paediatric home nursing team as described by Eaton (2000) are used to provide the framework with which to describe the service. A focus group held with the Kidz First paediatric community homecare nurses has enabled definition of the key nursing components provided to children and their families living in South Auckland. PMID- 12564522 TI - Stress, social support and psychological well-being in older men. AB - Along with an increase in the older population comes a concomitant increase in the number of people for whom issues relating to well-being, for example, stress and social relationships become more salient. In this study, relationships between stress, social support and psychological well-being in men over the age of 65 years were investigated. A convenience sample of 217 men over the age of 65 years, living in a small New Zealand city took part in the study. A questionnaire was used to gather information on stress, social support and psychological well being, as well as biographical data. Data were analysed quantitatively. Results showed that both the men who were satisfied with the social support they received and those who reported low stress levels experienced higher levels of psychological well-being (p<.001). The number of people available to provide social support was not significantly related to psychological well-being. There were no significant interaction effects between stress, social support variables and psychological well-being, with each variable operating independently in relation to psychological well-being. It appeared that the older the men were the fewer social supports available to them, suggesting the importance of the quality of those supports in maintaining psychological well-being. PMID- 12564523 TI - The Marquette Memorial Service for Nurses. PMID- 12564524 TI - A dream being realised. Report of the 13th congress of the International Council of Women's Health Issues held in Seoul, June 2002. PMID- 12564526 TI - Health survey of wild and captive bog turtles (Clemmys muhlenbergii) in North Carolina and Virginia. AB - Blood samples, fecal samples, and cloacal swabs were collected from 42 bog turtles (Clemmys muhlenbergii). including 14 wild males, 22 wild females, three captive males, and three captive females, in Virginia and North Carolina, USA. Samples were analyzed for hematologic and plasma chemistry values, Mycoplasma sp. antibodies, intestinal parasites, and normal cloacal flora. PMID- 12564527 TI - Paramyxovirus and reovirus infections in wild-caught Mexican lizards (Xenosaurus and Abronia spp.). AB - Virus isolation attempts were carried out on wild-caught Xenosaurus grandis, X. platyceps, and Abronia graminea from Mexico. These animals were also tested for exposure to paramyxoviruses and reoviruses. Pharyngeal and cloacal swabs were collected from 30 lizards, and blood was collected from 23 lizards. A cytopathogenic virus was isolated from the cloacal swab of one of the X. platyceps. The isolate was identified as a paramyxovirus on the basis of its sensitivity to chloroform, resistance to 5-iodo-2'-deoxyuridine, size and morphology of the viral particles, hemagglutination of chicken erythrocytes, and serologic reaction with paramyxovirus-specific antisera. Antibodies against the paramyxovirus isolated in this study were found in four animals from three species. Antibodies against a different paramyxovirus isolated from a monitor lizard were found in seven animals from three species, showing that all the species tested are susceptible to paramyxovirus infections. Antibodies to a reptilian reovirus were found in three of the X. grandis. PMID- 12564525 TI - Osteomyelitis associated with Salmonella enterica SS arizonae in a colony of ridgenose rattlesnakes (Crotalus willardi). AB - The identification of three Arizona ridgenose rattlesnakes (Crotalus willardi) with Salmonella arizonae-associated osteomyelitis led to a 5-yr prospective study of radiographic signs and Salmonella intestinal carriage rates in a 19-member colony of this rattlesnake species. Ventrodorsal radiographs were performed and cloacal swabs were cultured for Salmonella spp. annually. Ten snakes survived the 5-yr period, with six of them remaining free of bony lesions. Three snakes that had no bony lesions in 1995 developed radiographic signs of osteomyelitis during the study. Six snakes with bony lesions at the beginning of the study died or were euthanatized due to osteomyelitis during the study. The radiographic signs of osteomyelitis were progressive for five snakes that were serially radiographed. Only one snake with radiographic signs of osteomyelitis at the beginning of the study was still alive at the end of the study, and this animal's bony lesions were more extensive at the end. Thirty-nine intestinal S. arizonae isolates, representing 13 serotypes, were obtained from the 19 snakes. Salmonella arizonae serotype 56:Z4,Z23 was isolated only once from a cloacal culture, from a snake that had no radiographic bone lesions. Twelve extraintestinal Salmonella isolates, representing two serotypes, were isolated from six snakes. All extraintestinal isolates except one were of S. arizonae serotype 56:Z4,Z23, and all isolates from bone were of this serotype. One snake with characteristic bone lesions died, and Providencia rettgeri was cultured from each of the tissues cultured, whereas no Salmonella spp. were isolated from this snake. Salmonella arizonae serotype 56:Z4,Z23 [corrected] appears to have a tropism for bone and other extraintestinal sites in C. willardi and may cause a progressive, ultimately fatal disease in this species. PMID- 12564528 TI - The presence of fungi associated with sick building syndrome in North American zoological institutions. AB - A total of 110 sites from five zoological institutions were examined to determine whether fungi associated with sick building syndrome (SBS) were prevalent in the exhibits or night-time holding facilities and to investigate whether the presence of these organisms was associated with declining breeding rates or increases in morbidity and mortality (or both). Each site was sampled with an Andersen two stage air sampler using Sabourauds dextrose agar media and a Burkard personal volumetric air sampler. Suspect surfaces were also sampled. High levels of airborne Penicillium chrysogenum, a fungal species associated with poor indoor air quality, were recovered from 16 sites out of all five institutions. Five culturable growth sites of Stachybotrys chartarum, a species strongly associated with SBS and commonly known as "black mold," were recovered from surfaces at two institutions. A wide range of other fungal species was recovered in low numbers from all institutions. A Fisher exact test analysis showed a significant nonrandom association between high levels of P. chrysogenum and sites with records of poor animal health. This study indicated that significant numbers of airborne fungi associated with SBS and poor indoor air quality are present in zoological institutions and that they could affect animal health and reproduction rates and zoo staff. PMID- 12564529 TI - Hematology and serum chemistry values of the lesser rhea (Pterocnemia pennata) raised in Patagonian farms (Argentina). AB - Hematologic and serum chemistry values were determined for chicks, juveniles, and adult farmed lesser rheas (Prerocnemia pennata). Blood samples were taken during March-April 1998 from the brachial veins of lesser rhea, including 64 chicks, 24 juveniles, and 36 adults, raised in Patagonian farms (Argentina). Lesser rhea chicks had significantly lower erythrocyte counts, hematocrit, hemoglobin, red cell indices, and copper than did the juveniles and adult individuals. No significant differences were observed between females and males. Most values were similar to those reported for other ratites. The data obtained provide hematologic and serum chemistry values for lesser rhea from farms. PMID- 12564530 TI - Tiletamine-zolazepam, ketamine, and xylazine anesthesia of captive cheetah (Acinonyx jubatus). AB - Thirty-two anesthetic episodes used a combination of tiletamine-zolezepam (50 mg/ml each), ketamine (80 mg/ml), and xylazine (20 mg/ml) at various dosages for routine diagnostic and minor surgical procedures in 13 captive cheetahs (Acinonyx jubatus). The mean dosage (0.023 +/- 0.003 ml/kg) provided rapid induction with a single i.m. injection along with safe predictable working time, good muscle relaxation, and analgesia. Yohimbine administration subsequently accelerated smooth and rapid recovery. PMID- 12564531 TI - Sero-epizootiologic survey of Trypanosoma brucei in Kenyan nonhuman primates. AB - Blood samples were collected from 121 individuals of three species of wild-caught nonhuman primates from Kenya, including African green monkeys (Cercopithecus aethiops), Syke's monkeys (C. mitis), and olive baboons (Papio cynocephalus anubis), and were examined for circulating Trypanosoma brucei and for T. brucei antigen and anti-trypanosome antibody. Indirect antibody enzyme-linked immunosorbent assay detected titers of anti-T. brucei antibodies in 13 of the primates sampled, and field-oriented latex agglutination test detected invariant T. brucei antigens in 10 (8.3%) of the primates. However, no trypanosomes were visible in blood smears, on wet blood films, or by buffy coat technique, nor were they demonstrable in a subset of C. aethiops individuals that were studied using mouse subinoculation. PMID- 12564533 TI - Congenital diffuse hyperplastic goiter associated with perinatal mortality in 11 captive-born bottlenose dolphins (Tursiops truncatus). AB - Diffuse hyperplastic goiter was diagnosed by histopathology in 11 perinatal bottlenose dolphins (Tursiops truncatus) that died at four separate zoos and aquaria. Thyroid morphology of these animals was compared with the histologically normal thyroids of two stranded wild bottlenose dolphin calves, a neonate and a 2 mo-old calf. Histologic changes included reduced follicular luminal diameter, markedly reduced or absent luminal colloid, hypertrophy of follicular epithelium, and follicular dysplasia. The etiology of the thyroid gland lesion was not identified. Cause of death was not determined for most of these animals, but they were presumed to have died from metabolic derangements associated with the thyroid lesion, drowning, or dystocia. PMID- 12564532 TI - Surgical implantation of radio transmitters in arctic foxes (Alopex lagopus) on Svalbard, Norway. AB - Twelve captive wild-caught adult arctic foxes (Alopex lagopus) were anesthetized a total of 24 times for an equal number of surgical procedures involving implantation of heart rate (HR) and core body temperature transmitters (Tb) between October 1995 and April 1997. Xylazine-ketamine and medetomidine-ketamine anesthesia was used, resulting in an unacceptably high death rate. One out of four foxes anesthetized with xylazine-ketamine died, whereas two of nine foxes anesthetized with medetomidine-ketamine died out of a total of 20 surgical procedures. Durations of the surgeries for implantation of Tb transmitters and HR transmitters were 73 +/- 7 min and 95 +/- 13 min, respectively. PMID- 12564535 TI - Lingual squamous cell carcinoma in a California sea lion (Zalophus californianus). AB - A 28-yr-old female California sea lion (Zalophus californianus) in a commercial aquarium developed an ulcerated lingual tumor and died. Necropsy revealed a moderately differentiated squamous cell carcinoma. Immunohistochemical staining and electron microscopic studies revealed that the tumor cells were strongly positive with anti-keratin-cytokeratin antibody and had abundant tonofibrils and desmosomes. The neoplasm had metastasized to a mandibular lymph node. PMID- 12564534 TI - Saksenaea vasiformis and Apophysomyces elegans zygomycotic infections in bottlenose dolphins (Tursiops truncatus), a killer whale (Orcinus orca), and pacific white-sided dolphins (Lagenorhynchus obliquidens). AB - During a 10-yr period, a killer whale (Orcinus orca), two Pacific white-sided dolphins (Lagenorhynchus obliquidens), and two bottlenose dolphins (Tursiops truncatus), all housed at SeaWorld of Texas from 1991 to 2001, were infected with fungi from the class Zygomycetes. In four out of five cases, the fungi were identified as either Saksenaea vasiformis or Apophysomyces elegans. All fungi in the class Zygomycetes aggressively invade the vascular system. Death occurred within 23 days after the initial clinical signs. The primary site of infection involved the s.c. tissue and skeletal musculature. In one case, infection originated in the placenta and uterus of a periparturient animal. All cases exhibited systemic spread of the organisms, including two to the central nervous system. The fifth and most recent case, a bottlenose dolphin, was treated with liposomal nystatin, an antifungal formulation with reduced nephrotoxicity. This animal initially responded to therapy; however, 14 days after cessation of therapy, fungal growth reoccurred. Thus, the animal was euthanatized 39 days after the initial clinical signs. This drug represents a promising treatment option if combined with early disease detection and aggressive tissue resection. PMID- 12564536 TI - Nasopharyngeal myxosarcoma in a Bengal tiger (Panthera tigris). AB - An 18- to 25-yr-old intact female Bengal tiger (Panthera tigris) was diagnosed antemortem and postmortem with nasopharyngeal myxosarcoma metastatic to the lung, chronic lymphoplasmacytic otitis media, and lymphoplasmacytic and eosinophilic stomatitis. Myxosarcomas are rare in domestic animals and seldom metastasize; this tumor has not been previously reported in an exotic felid. Computed tomography of the skull was used during the diagnoses. PMID- 12564537 TI - Survey of tuberculin testing in Swedish zoos. AB - Tuberculin test results from 214 animals in three Swedish zoos, tested between the years 1993 and 2000, were compiled from a questionnaire sent out to zoo veterinarians. Comparative testing with bovine and avian tuberculin was used on various sites of injection. A total of five skin test reactors were found: three cotton-top tamarins (Saguinus oedipus) in one zoo and two tapirs (Tapirus terrestris) in another zoo. Postmortem culture from one of the tapirs revealed growth of Mycobacterium tuberculosis, and a stamping out policy was adopted in the herd. Tuberculosis in the primates was ruled out by further investigations. Zoo veterinarians should try to adopt a common scheme for the regular testing of zoo animals to improve the diagnostic ability and comparison of results between institutions. PMID- 12564538 TI - Cerebral arteriosclerosis in an aged captive polar bear (Ursus maritimus). AB - Cerebral arteriosclerosis was observed upon necropsy of a 36-yr-old female captive polar bear (Ursus maritimus) that developed a sudden onset of seizure like activity and died. The medium and large cerebral arteries of the meninges had moderate to severe diffuse discoloration and mineralization of the matrix of the tunica media, with little or no associated cellular reaction. Scanning electron microscopy of the affected arteries showed discrete crystalline calcified deposits in the media and sclerosis of the arterial wall. There were no lesions in the brainstem. The findings suggested a sudden and rapidly fatal loss of blood flow to the brain caused by long-standing arterial lesions. Incidental findings included numerous 0.1- to 10-cm-diameter, hepatic cysts lined with hyperplastic biliary epithelium, a unilateral, unipolar, 3-cm-diameter renal tubular adenoma, and approximately 250 active Baylisascaris sp. nematodes in the intestines. PMID- 12564539 TI - Persistent right aortic arch in a yearling captive wood bison (Bison bison athabascae). AB - An undersized, 16-mo-old, captive wood bison (Bison bison athabascae) was observed with dysphagia and postprandial regurgitation. The animal died shortly after anesthesia was initiated for diagnostic purposes. At necropsy, a persistent right aortic arch and aberrant left subclavian artery were diagnosed. The vascular ring structure around the esophagus and the trachea caused a megaesophagus with complete esophageal obstipation and ulcerative esophagitis. The aberrant left subclavian artery did not significantly compromise the esophagus. Persistence of the right aortic arch is relatively rare in ruminants and has not previously been reported in a bison. PMID- 12564541 TI - Thyroid c-cell carcinoma in an African pygmy hedgehog (Atelerix albiventris). AB - A 3-yr-old African pygmy hedgehog (Atelerix albiventris) was submitted with dysphagia, weight loss, and tetraparesis. A palpable mass was found on the ventral neck. Histologic examination revealed replacement of the thyroid gland by a highly cellular, expansile, and infiltrative mass composed of lobules of polygonal cells separated by fine fibrovascular septa. Examination of ultrathin sections revealed tumor cells with few to many dense-core neuroendocrine granules, approximately 100-200 nm in diameter, and stromal amyloid. Immunohistochemical stains were positive for neuron-specific enolase. Only rare cells had positive immunohistochemical staining for calcitonin. Findings are consistent with a neuroendocrine tumor of C-cell origin. This is the first report of a C-cell carcinoma in a hedgehog. PMID- 12564540 TI - Combination therapy for footpad lesions in a captive Bengal tiger (Panthera tigris tigris). AB - A captive adult male Bengal tiger (Panthera tigris tigris) was treated for chronic footpad ulcers associated with pacing on concrete cage surfaces. Combination therapy of oral fluoxetine and acepromazine administration to diminate pacing behavior, daily application of moisturizing ointment, and oral vitamin E supplementation resulted in complete resolution of all footpad lesions. Subsequent clinical episodes completely resolved with similar treatments. PMID- 12564542 TI - Medetomidine-ketamine in reindeer (Rangifer tarandus tarandus): effective immobilization by hand- and dart-administered injection. AB - Twelve reindeer (Rangifer tarandus tarandus) were immobilized by hand injection in indoor stalls with established optimal hand-injection doses of medetomidine ketamine and then moved to outside paddocks where they were immobilized again with the same dose by dart. The reindeer in paddocks were immobilized a second time with a 50% higher dose, hereafter referred to as the optimal darting dose. Mean time to first sign of sedation was longer and mean induction time was significantly longer (55% and 79%, respectively) when the optimal hand-injection dose was dart injected versus hand injected. Mean time to first sign of sedation was not significantly shorter (although 21% shorter, numerically) but mean induction time was significantly shorter (30%) when animals were darted with the optimal darting dose versus darted with the optimal hand-injection dose. There were no significant differences in respiratory rate, rectal temperature, and relative arterial oxygen saturation in animals injected with different doses and by different routes. but there was a significantly lower heart rate in animals dart injected with the optimal darting dose versus dart injected with the optimal hand-injection dose. All animals responded at similar rates to atipamezole injection. PMID- 12564543 TI - Individually coded telemetry: a tool for studying heart rate and behaviour in reindeer calves. AB - The aim of the study was to test the performance of a silver wire modified version of the coded telemetric heart rate monitor Polar Vantage NV (PVNV) and to measure heart rate (HR) in a group of captive reindeer calves during different behaviour. The technical performance of PVNV HR monitors was tested in cold conditions (-30 degrees C) using a pulse generator and the correlation between generated pulse and PVNV values was high (r=0.9957). The accuracy was tested by comparing the HR obtained with the PVNV monitor with the standard ECG, and the correlation was significant (r=0.9965). Both circadian HR and HR related to behavioural pattern were recorded. A circadian rhythm was observed in the HR in reindeer with a minimum during night and early morning hours and maximum at noon and during the afternoon, the average HR of the reindeer calves studied being 42.5 beats/min in February. The behaviour was recorded by focal individual observations and the data was synchronized with the output of the HR monitors. Running differed from all other behavioural categories in HR. Inter-individual differences were seen expressing individual responses to external and internal stimuli. The silver wire modified Polar Vantage NV provides a suitable and reliable tool for measuring heart rate in reindeer, also in natural conditions. PMID- 12564544 TI - Incidence of reinfections with Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae in pig farms located in respiratory-disease-free regions of Switzerland--identification and quantification of risk factors. AB - The objective of the study was to identify risk factors for reintroduction of Actinobacillus pleuopneumoniae and Mycoplasma hyopneumoniae (enzootic pneumonia) onto pig farms in areas in Switzerland that were involved in an eradication programme from 1996 to 1999 and to assess the role of dealers in relation to these reinfections. The study was based on the comparison of pig farms that were reinfected in the year 2000 (cases) and pig farms that remained uninfected in the same area (controls). Additionally, data were collected from Swiss pig dealers and transport companies. Out of a total of 3983 farms, 107 farms were reinfected in the year 2000. The incidences were 0.1% for Actinobacillus pleuopneumoniae and 2.6% for Mycoplasma hyopneumoniae (enzootic pneumonia). Compared to reinfection rates prior to the eradication programme, this is a considerable reduction. Statistically significant risk factors for the reinfection were 'finishing farm', 'large mixed breeding-finishing farm', 'reinfected neighbour' and 'parking site for pig transport vehicles close to the farm'. Pig farmers that purchased pigs from only one supplier per batch had a lower risk of reintroducing infection (protective factor). As long as infected and uninfected regions co-exist in Switzerland, direct and indirect contact between farms, pig herds and slaughter sites via transport vehicles are a major pathway of disease spread. Risk management measures linked to these contacts are therefore of key importance. The survey of dealers indicated various areas for improvement such as strategic planning of pick-up routes or cleaning and disinfecting of trucks. PMID- 12564545 TI - Oestrone sulphate measurements for the prediction of small or large litters in pigs. AB - Serum from 88 pregnant sows and gilts was sampled 24 and 28 days after their first insemination or mating day. The oestrone sulphate (E1S) concentration in the samples was assessed with a commercially available radioimmunoassay kit modified for use with swine serum. The first aim was to test whether it was possible to predict litters of total number <10 piglets at term. The second aim was to compare the use of day 24 or day 28 samples, or of both, in this prediction. Day 24 E1S levels were positively correlated with litter size at term (R2 = 0.26; p <0.001). E1S levels on day 28 were correlated with the levels on day 24 in the same animals but could not be used for prediction of large or small litters. The odds ratio for a small litter size was 0.16 (p <0.01). This means that odds for a litter size <10 piglets decreased by 84% when E1S levels increased by 1.0 ng/ml. PMID- 12564547 TI - Clinical parameters for assessment of udder health in Danish dairy herds. AB - This study examined the possibilities of using clinical parameters related to the bovine udder for characterisation of udder health. Five clinicians performed systematic clinical recordings of udder health at 3 visits to 4 dairy herds. Several of the clinical parameters were scored on an ordinal scale. The agreement between clinicians was compared using kappa statistics. Factor analysis was used to identify udder types. The clinical evaluations showed substantial variation among clinicians. Parameters that were not directly related to pathological conditions showed the highest variation e.g. length of the claws, teat shape and hardness of the udder parenchyma. On the other hand, evaluation of pathological parameters such as nodes in the udder, skin lesions and oedema showed good agreement between clinicians. Udder types identified by means of factor analysis were found to be suitable for characterisation of udder health. Especially one factor related to dry quarters and udder asymmetry showed a more consistent relationship to milk yield than traditionally applied udder health parameters such as treatment rate and cell count. It is concluded that there is a considerable need for increased efforts among clinicians in order to standardise clinical recordings. It is further concluded that certain combinations of extended clinical recordings have significant perspectives for future characterisation of udder health. PMID- 12564546 TI - Immune-mediated fever in the dog. Occurrence of antinuclear antibodies, rheumatoid factor, tumor necrosis factor and interleukin-6 in serum. AB - Contents of antinuclear antibodies (ANA), rheumatoid factor (RF), tumor necrosis factor (TNF-alpha) and interleukin-6 (IL-6) were measured in serum from 20 dogs with immune-mediated fever. Seven out of 20 patients were ANA positive, 1 out of 20 was positive to antibodies against extractable nuclear antigens (ENA), 1 out of 20 was positive to antibodies against deoxynucleoproteins (DNP), 2 out of 13 were RF positive and none out of 20 patients had antibodies against native DNA in the serum. TNF-alpha was not detected in any serum of 15 dogs with immune mediated fever, while 10 out of 13 presented with elevated IL-6. The results varied between patients, but the IL-6 level was high in most of them. This indicate a role for IL-6 in the pathogenesis of immune-mediated fever in most cases. PMID- 12564548 TI - A method for the recovery of immature Trichuris suis from pig intestine. PMID- 12564549 TI - Mycoplasma hyorhinis isolation from cases of otitis media in piglets. PMID- 12564550 TI - A breadth-first survey of eye-tracking applications. AB - Eye-tracking applications are surveyed in a breadth-first manner, reporting on work from the following domains: neuroscience, psychology, industrial engineering and human factors, marketing/advertising, and computer science. Following a review of traditionally diagnostic uses, emphasis is placed on interactive applications, differentiating between selective and gaze-contingent approaches. PMID- 12564551 TI - A contingent speech technique in eye movement research on reading. AB - A novel eye-movement-contingent method is presented. It builds on and extends established eye-movement-contingent visual display change methods in that it uses movements of the eyes to control the presentation of acoustic information during sentence reading. In one implementation, an irrelevant spoken word is presented when the eyes cross a predetermined spatial boundary before they move on to a selected visual target word. The relationship between the spoken word and the visual target is manipulated, and the pattern of interference, caused by the presentation of the spoken word, is used to determine the nature and time course of activated representations. Results from three recently completed experiments in which the technique was used show that a word's phonological code remains active after it has been read and that the activated code has speech-like properties. PMID- 12564552 TI - Perception onset time during fixations in free viewing. AB - In this study, we investigated when visual perception begins in fixations. During picture viewing, the picture was degraded at the beginning of selected saccades and changed back to the original after varying intervals. Participants manually responded whenever they detected changes. The change-backs were undetected when they occurred <6 msec after the end of the saccade, marked by the peak of the overshoot in dual Purkinje image eyetracker data, and detection reached asymptote 32 msec after that marker. Eye velocity at the change-back time also affected detection likelihood. Apparently, perception begins around the time at which the eyes stop rotating at the end of a saccade, giving a psychological justification for measuring fixation durations from then. This also specifies the deadline for gaze-contingent display changes to occur without detectable image motion. Investigators using the dual Purkinje image eyetracker should consider the peak of the overshoot as the fixation onset time and measure intrafixational presentation times from then. PMID- 12564553 TI - Saliency of peripheral targets in gaze-contingent multiresolutional displays. AB - Gaze-contingent multiresolutional displays (GCMRDs) have been proposed to solve the processing and bandwidth bottleneck in many single-user displays, by dynamically placing high-resolution in a window at the center of gaze, with lower resolution everywhere else. The three experiments reported here document aslowing of peripheral target acquisition associated with the presence of a gaze contingent window. This window effect was shown for displays using either moving video or still images. The window effect was similar across a resolution-defined window condition and a luminance-defined window condition, suggesting that peripheral image degradation is not a prerequisite of this effect. The window effect was also unaffected by the type of window boundary used (sharp or blended). These results are interpreted in terms of an attentional bias resulting in a reduced saliency of peripheral targets due to increased competition from items within the window. We discuss the implications of the window effect for the study of natural scene perception and for human factors research related to GCMRDs. PMID- 12564554 TI - Using MATLAB with the psychophysics toolbox to present the heterochromatic fusion nystagmus stimulus. AB - We present a program for MATLAB that generates and presents the heterochromatic fusion nystagmus stimulus. This stimulus allows assessment of isoluminant states through recordings of reflexive eye movements (optokinetic nystagmus). The reflexive nature of the subject's response makes this stimulus especially useful with nonverbal subjects, such as children and animals. Unfortunately, the stimulus is complex and difficult to program. By presenting the present program, we hope to help those who wish to use this tool in their research. The code of the function can be downloaded at www. dal.ca/-mcmullen/downloads.html. PMID- 12564555 TI - Eye movements of large populations: I. Implementation and performance of an autonomous public eye tracker. AB - This paper details the design and construction of an autonomous public eye tracker exhibit, which was installed at the National Gallery, London, in 2000/2001. For over 3 months, it functioned both as an informative exhibit and as a controlled eye movement experiment, gathering data from over 5,000 participants. The issues associated with automatic unattended recording of the eye movements of members of the public are discussed. The performance of the exhibit is examined, and its successes and problem areas are highlighted with regard to potential applications and future exhibits. The success of the project proves the viability of autonomous public eye trackers as both data-gatherers and public exhibits. PMID- 12564556 TI - Eye movements of large populations: II. Deriving regions of interest, coverage, and similarity using fixation maps. AB - The analysis of eye movement traces (i.e., the patterns of fixations in a search) is more complex than that of such parameters as mean fixation duration, and as a result, previous attempts have focused on a qualitative appraisal of the form of an eye movement trace. In this paper, the concept of the fixation map is introduced. Its application to the quantification of similarity of traces and the degree of coverage by fixations of a visual stimulus is discussed. The ability of fixation maps to aid in the understanding and communication of large numbers of eye movement traces is examined. PMID- 12564557 TI - Glance analysis of driver eye movements to evaluate distraction. AB - With the increasing use of in-vehicle devices in cars, an understanding of the safety implications of secondary tasks has become crucial. It is now possible to study the effects of many in-vehicledevices and tasks on driving by using head mounted eye-tracking devices (HEDs) to collect eye positions and pupil diameters, which have been considered indicators of attentional focus. The collection of eye position and pupil-diameter data of automobile drivers under on-road conditions and while completing various secondary tasks is described in this paper. Drivers were asked to drive on a preselected two-lane road for a total distance of 22 miles while gaze data were recorded using a HED. Longer off-road fixation durations were observed in radio-tuning and rearview mirror checking tasks, but not in the odometer checking task. In addition, the standard deviations of fixation displacements during a cognitive task involving the computation of a date for a meeting were shorter than those observed during normal driving. PMID- 12564558 TI - Attention to repeated images on the World-Wide Web: another look at scanpath theory. AB - The scanpath theory of visual perception was tested using Web pages as visual stimuli. Scanpaths are repetitive sequences of fixations and saccades that occur upon reexposure to a visual stimulus. Since Internet users are exposed to repeated visual displays, the Web provides ideal stimuli to test this theory. Eye movement data were recorded for subjects' repeated viewings of three Web pages over three sessions. Resemblance of eye path sequences was measured with a string edit method; multidimensional scaling and cluster analysis were used to group sequences. Support was found for the scanpath theory; some clusters included pairs of sequences from the same subject. A repeated measures analysis of variance revealed a statistically significant main effect for stimulus type, with a text-intensive news story page generating more similar sequences than a graphic intensive advertising page. There was a statistically significant main effect for cross-viewing comparisons, reflecting a linear trend in which eye paths for the same subject became more alike over time. PMID- 12564559 TI - Using high frame rate CMOS sensors for three-dimensional eye tracking. AB - A novel three-dimensional eye tracker is described and its performance evaluated. In contrast to previous devices based on conventional video standards, the present eye tracker is based on programmable CMOS image sensors, interfaced directly to digital processing circuitry to permit real-time image acquisition and processing. This architecture provides a number of important advantages, including image sampling rates of up to 400/sec measurement, direct pixel addressing for preprocessing and acquisition,and hard-disk storage of relevant image data. The reconfigurable digital processing circuitry also facilitates inline optmization of the front-end, time-critical processes. The primary acquisition algorithm for tracking the pupil and other eye features is designed around the generalized Hough transform. The tracker permits comprehensive measurement of eye movement (three degrees of freedom) and head movement (six degrees of freedom), and thus provides the basis for many types of vestibulo oculomotor and visual research. The device has been qualified by the German Space Agency (DLR) and NASA for deployment on the International Space Station. It is foreseen that the device will be used together with appropriate stimulus generators as a general purpose facility for visual and vestibular experiments. Initial verification studies with an artificial eye demonstrate a measurement resolution of better than 0.1 degrees in all three components (i.e.,system noise for each of the components measured as 0.006 degrees H, 0.005 degrees V, and 0.016 degrees T. Over a range of +/-20 degrees eye rotation, linearity was found to be <0.5% (H), <0.5% (V), and <2.0% (T). A comparison with the scleral search coil technique yielded near equivalent values for the system noise and the thickness of Listing's plane. PMID- 12564560 TI - A laser-based eye-tracking system. AB - This paper reports on the development of a new eye-tracking system for noninvasive recording of eye movements. The eye tracker uses a flying-spot laser to selectively image landmarks on the eye and, subsequently, measure horizontal, vertical, and torsional eye movements. Considerable work was required to overcome the adverse effects of specular reflection of the flying-spot from the surface of the eye onto the sensing elements of the eye tracker. These effects have been largely overcome, and the eye-tracker has been used to document eye movement abnormalities, such as abnormal torsional pulsion of saccades, in the clinical setting. PMID- 12564561 TI - 3-D eye movement analysis. AB - This paper presents a novel three-dimensional (3-D) eye movement analysis algorithm for binocular eye tracking within virtual reality (VR). The user's gaze direction, head position, and orientation are tracked in order to allow recording of the user's fixations within the environment. Although the linear signal analysis approach is itself not new, its application to eye movement analysis in three dimensions advances traditional two-dimensional approaches, since it takes into account the six degrees of freedom of head movements and is resolution independent. Results indicate that the 3-D eye movement analysis algorithm can successfully be used for analysis of visual process measures in VR. Process measures not only can corroborate performance measures, but also can lead to discoveries of the reasons for performance improvements. In particular, analysis of users' eye movements in VR can potentially lead to further insights into the underlying cognitive processes of VR subjects. PMID- 12564562 TI - Cleaning up systematic error in eye-tracking data by using required fixation locations. AB - In the course of running an eye-tracking experiment, one computer system or subsystem typically presents the stimuli to the participant and records manual responses, and another collects the eye movement data, with little interaction between the two during the course of the experiment. This article demonstrates how the two systems can interact with each other to facilitate a richer set of experimental designs and applications and to produce more accurate eye tracking data. In an eye-tracking study, a participant is periodically instructed to look at specific screen locations, or explicit required fixation locations (RFLs), in order to calibrate the eye tracker to the participant. The design of an experimental procedure will also often produce a number of implicit RFIs--screen locations that the participant must look at within a certain window of time or at a certain moment in order to successfully and correctly accomplish a task, but without explicit instructions to fixate those locations. In these windows of time or at these moments, the disparity between the fixations recorded by the eye tracker and the screen locations corresponding to implicit RFLs can be examined, and the results of the comparison can be used for a variety of purposes. This article shows how the disparity can be used to monitor the deterioration in the accuracy of the eye tracker calibration and to automatically invoke a recalibration procedure when necessary. This article also demonstrates how the disparity will vary across screen regions and participants and how each participant's unique error signature can be used to reduce the systematic error in the eye movement data collected for that participant. PMID- 12564563 TI - ILAB: a program for postexperimental eye movement analysis. AB - The recording and analysis of eye movements are fundamental to a diverse set of research applications, including studies in which reading, visual search, and both overt and covert visuospatial attention are examined. Software tools supplied with commonly available eye-tracking equipment have generally been limited in functionality and nonextensible. Because of this dearth of available software, ELAB was created to provide an extensible framework for analyzing various aspects of eye movements. The program consists of a series of open-source MATLAB functions. The program's data structures keep raw data, analysis preferences, and analyzed data separate, thus maintaining data fidelity and promoting extensibility. PMID- 12564564 TI - The Eyelink Toolbox: eye tracking with MATLAB and the Psychophysics Toolbox. AB - The Eyelink Toolbox software supports the measurement of eye movements. The toolbox provides an interface between a high-level interpreted language (MATLAB), a visual display programming toolbox (Psychophysics Toolbox), and a video-based eyetracker (Eyelink). The Eyelink Toolbox enables experimenters to measure eye movements while simultaneously executing the stimulus presentation routines provided by the Psychophysics Toolbox. Example programs are included with the toolbox distribution. Information on the Eyelink Toolbox can be found at http://psychtoolbox.org/. PMID- 12564565 TI - Measuring saccade curvature: a curve-fitting approach. AB - Saccade curvature is becoming a popular measure for detecting the presence of competing saccadic motor programs. Several different methods of quantifying saccade curvature have been employed. In the present study, we compared these metrics with each other and with novel measures based on curve fitting. Initial deviation metrics were only moderately associated with the more widely used metric of maximum curvature. The latter was strongly related to a recently developed area-based measure and to the novel methods based on second- and third order polynomial fits. The curve-fitting methods showed that although most saccades curved in only one direction, there was a population of trajectories with both a maximum and a minimum (i.e., double-curved saccades). We argue that a curvature metric based on a quadratic polynomial fit deals effectively with both types of trajectories and, because it is based on all the samples of a saccade, is less susceptible to sampling noise. PMID- 12564566 TI - A turn-key transportable eye-tracking instrument for clinical assessment. AB - This paper describes a combined instrument (eye tracker and target generator, both head mounted, with integrated data analysis) that tests parameters of saccadic eye movement and fixation control to give insight into the status of functional brain systems. Using three minilasers, the target generato rprojects three visual stimuli, a fixation point and two lateral stimuli, with programmable timing. The controller allows the selection of overlap, 200-msec gap, or remembered saccade trials. Size, maximal velocity, and reaction time are determined for each primary saccade. The number of prosaccades and antisaccades are counted. More saccades--for example, the occurrence and latency of corrective saccades--may be evaluated off line by an interactive PC analysis program. The eye position data can be transferred to a PC. Off-line analysis compares each observed variable relative to an age-matched control group (300 healthy control subjects 7-70 years of age, tested in the overlap condition with prosaccade instructions and in the gap condition with antisaccades). The diagnostic results can be used to elaborate an individual optomotor training program. PMID- 12564567 TI - The general practitioner needs to be made better known. PMID- 12564568 TI - General practitioners' continuing medical education: a prospective study from the County of Aarhus. AB - Participation of Danish general practitioners (GPs) in continuing medical education (CME) has often been the subject of debate, although very little is known about the extent and content of activities. One-hundred-and-sixty-one Danish GPs participated in this one-year prospective study by collecting data on their own CME activities. We received 9980 data registration charts. During an average period of 8.4 months, the average Danish GP spent 67 h on traditional CME, equivalent to approximately 96 h per year, and 12 h on small group-based CME per year. In addition, he/she spent 90 h per year reading textbooks, journals, etc. The time spent on CME therefore totalled more than 200 h per year. Most of the CME courses took place outside surgery hours. We conclude that Danish GPs participate in a large number of CME activities--even more than their own organisation recommend. PMID- 12564569 TI - Patients' and doctors' views on respiratory tract symptoms. AB - OBJECTIVE: To explore the views of patients and doctors on respiratory tract symptoms. DESIGN: Transversal survey among patients and general practitioners. SETTING: Primary health care and community. SUBJECTS: 51 patients attending a general practitioner, 38 patients in the community, 7 general practitioners. MAIN OUTCOME MEASURES: Patients' and doctors' views on respiratory tract symptoms and differences between them: agreement with statements rated on a 5-point scale, ranging from "strongly disagree" to "strongly agree." RESULTS: Patients less than doctors endorsed the self-limiting character of cough, sore throat and earache (mean 3.1, 3.4 and 2.9 versus 4.1, 4.1 and 3.7) and patients--much more than doctors--rated antibiotics as being necessary for cough and sore throat (mean 2.7 and 2.9 versus 1.7 and 17) and believed that antibiotics speed recovery (mean 3.7 versus 2.0). However, there was little difference relating to the necessity to see a doctor after some time period. CONCLUSION: Patients appeared to differ from doctors in views on respiratory tract symptoms. The results stress the importance of exploring patients' views when being confronted by patients suffering from respiratory tract symptoms. PMID- 12564570 TI - GPs and lesbian women in the consultation: issues of awareness and knowledge. AB - OBJECTIVE: To study awareness of having lesbian women in the consultation among general practitioners (GPs) and their knowledge of lesbian health-related issues. DESIGN: A mailed questionnaire study using closed and open-ended questions, with an analysis using both quantitative and qualitative methods. SETTING: The city of Goteborg, Sweden. SUBJECTS: GPs working in the city of Goteborg. MAIN OUTCOME MEASURES: Frequencies are presented for closed questions. Open-ended questions are categorised and illustrated by citations. RESULTS: The response rate was 52%, but only 37% were aware of having had any lesbian patients despite many years in practice. Five per cent had ever asked their patients about sexual identity, and most questions concerning social network were put in terms of the heterosexual, nuclear family. Eleven per cent knew of any health issues relevant to lesbian women, but half of all informants declared an interest in learning more. CONCLUSION: Unreflected assumptions of heterosexuality and use of heterosexist concepts may work together in keeping the lesbian patient invisible to health care. Medical education needs to include issues of gender and sexual identity/orientation in the curriculum, and to address the health effects of marginalisation. PMID- 12564571 TI - Care by general practitioners and district nurses of patients receiving home nursing: a study from suburban Stockholm. AB - OBJECTIVES: To review the care by general practitioners (GPs), district nurses and assistant nurses of patients receiving home nursing. DESIGN: Retrospective data from questionnaires, records and official statistics. Statistical comparisons. SETTING: Primary health care from October 1995 to October 1996. SUBJECTS: One-third (158) of all patients receiving home nursing in a suburban area were sampled; 73% (116) participated. All patients of comparable age in one practice served as a control group. MAIN OUTCOME MEASURES: Number of and reasons for visits and other contacts. Nature of care. Relation between patient problems and care given. RESULTS: Most patients were seen by the nurses two to five times a month. They met their GPs less often than other patients. More measures were undertaken without direct contact between GP and patient. The most common measures concerned medication and the assessment of symptoms. Patients with cognitive problems seemed to get less active GP care. CONCLUSION: GPs played an active role in the care of patients receiving home nursing even though they seldom met them. Many patients were regularly assessed by the nurses, which might have diminished the need for doctor visits. The care of patients with cognitive problems needs further study. PMID- 12564572 TI - The Finnmark Intervention Study. Better health for the fishery population in an Arctic village in North Norway. AB - OBJECTIVE: To evaluate the lifestyle and cardiovascular risk factor changes after 6 years of intervention in the North Cape community. DESIGN: Longitudinal cohort study with a quasi-experimental design with one intervention and three control communities selected from the same coastal area with a baseline screening (1987), 6-year intervention, and re-screening (1993). SETTING: Fishing communities on the coast of Finnmark in the Norwegian Arctic area. PARTICIPANTS: 1685 (70%) of the invited in both screenings, aged between 20 and 62 years in 1987. INTERVENTION: Community-intervention based on empowerment and cooperation between voluntary organisations and local health services. MAIN OUTCOME MEASURES: Change in cardiovascular risk factors. RESULTS: Compared to the control communities, the main findings in North Cape were among men: 21.3% less drinking boiled coffee (p < 0.05), 10.5% more drinking low fat milk (p < 0.01), 0.3 mmol/l reduction in cholesterol (p < 0.01) and 0.2 kg/m2 reduction in BMI (p < 0.001). Among women, there were 5.9% fewer smokers (p < 0.01), 21% less drinking boiled coffee (p < 0.05), 1 kg/m2 less increase in BMI (p < 0.001) and 0.5 less increase in MI risk score (p < 0.05). CONCLUSIONS: Small close-knit communities are suitable for community-based interventions where it is easier to obtain close interaction between health service, voluntary organisations and the public at large. PMID- 12564573 TI - Cholesterol reduction following health screening in general practice. AB - OBJECTIVES: To evaluate changes in plasma cholesterol following health screening and health discussions in general practice. DESIGN: Randomised prospective population-based study conducted over a period of 5 years. SETTING: Primary care, all general practitioners (GPs) in a well-defined area. SUBJECTS: A random sample of inhabitants aged 30-49 years in January 1991, registered with a local GP was invited to participate. The participants (1507 persons, or 75.4% of the 2000 invited) were randomly allocated to two intervention groups and a control group. MAIN OUTCOME MEASURES: Plasma cholesterol, percentage of subjects with plasma cholesterol higher than 7 mmol/l. RESULTS: After 5 years of intervention, plasma cholesterol in the whole population was significantly lower in the intervention groups compared to the control group. The decrease was most pronounced (0.5 mmol/l) in subjects at high cardiovascular risk. The percentage of high-risk individuals with a cholesterol level higher than 7 mmol/l was significantly lower in the intervention groups compared to the control group (9.8% vs 6.2%, p = 0.04), corresponding to a 37% reduction. CONCLUSIONS: The study shows that the health checks had a measurable impact on plasma cholesterol levels, the most pronounced effect is seen among individuals at high cardiovascular risk. PMID- 12564575 TI - Metabolic control and quality of data in medical records for subjects with type 2 diabetes in Swedish primary care: improvement between 1995 and 2001. AB - OBJECTIVE: To compare the registering of diabetic complications and metabolic control in type 2 diabetic subjects in primary health care in 1995 and 2001. DESIGN: Cross-sectional surveys of medical records. SETTING: Two primary health care centres in Stockholm County. SUBJECTS: Diabetic patients aged 35-64 years; in 1995, 59 men and 42 women and in 2001, 80 men and 80 women. MAIN OUTCOME MEASURES: Rate of notified examinations in medical records, diabetic complications and metabolic control. RESULTS: Increased rates between 1995 and 2001 were found regarding examinations for neuropathy (46% vs 79%), albuminuria (74% vs 91%), micro-albuminuria (11% vs 59%), BMI (33% vs 66%), HbA1c (88% vs 99%), cholesterol (69% vs 92%) and LDL cholesterol (42% vs 89%) (p < 0.001). Increased rates of perceived treatment goals between 1995 and 2001 were found regarding HbA1c (< 6.5%) (28% vs 57%) and cholesterol (< 5.0 mmol/l) (23% vs 36%) (p < 0.001), and also regarding LDL cholesterol (30% vs 36%) (p < 0.01). In men, the number of subjects with any macrovascular complication decreased between 1995 and 2001 (34% vs 17%) (p < 0.01), and in women the number of subjects with coronary heart disease decreased (26% vs 6%) (p < 0.01). CONCLUSION: Documented examination rates and metabolic control improved between 1995 and 2001. PMID- 12564574 TI - Blood pressure and other cardiovascular risk factors among treated hypertensives in Swedish primary health care. AB - OBJECTIVE: To evaluate antihypertensive treatment and other cardiovascular risk factors in primary health care. DESIGN: Cross-sectional survey of consecutive patients with treated hypertension in 1999. SETTING: 17 primary care centres in Sweden. SUBJECTS: 512 patients (mean age 67; SD 11 years). MAIN OUTCOME MEASURES: Antihypertensive treatment, cardiovascular risk factors. RESULTS: Patients with high diastolic BP (> or = 100 mmHg) and systolic BP (> 180 mmHg) values were few. The proportions with diastolic BP < 90, BP < 160/95 and < 140/90 mmHg were 64%, 54% and 15%. Mono-therapy was given in 51%, and > or = 3 drugs in 13%. Hypertensives with hyperlipidaemia were 42%, and only 26% of them were given lipid-lowering drugs, mainly statins, 21%. Smokers were 10%, 23% had diabetes, and many had overweight BMI = 25 kg/m2, 72%. CONCLUSION: Although two-thirds had diastolic BP < 90 mmHg, few had BP below the current treatment target < 140/90 mmHg. More than half of the hypertensives had at least one additional cardiovascular risk factor, and these hypertensives also had low proportions within several current treatment targets of hypertension and hyperlipidaemia, implying a need for intensified multiple risk factor intervention. PMID- 12564576 TI - Drug prescribing in public primary care centres. Results from prescription studies 1988-1997 in the county of Halland, Sweden. AB - OBJECTIVE: To present the prescribing patterns of general practitioners (GPs) at public primary care centres (PPCCs) in Halland, a county in the south-west of Sweden. GP share of the total prescribing of different drug groups 1988-1997 is presented, as well as changes in patterns. DESIGN: A descriptive prescription study performed 3 months each year in 10 consecutive years. SETTING: Medical service and pharmacies in Halland. SUBJECTS: Prescriptions from about 100 GPs of PPCCs and 550 physicians of various other specialties. MAIN OUTCOME MEASURES: Percentages and absolute numbers of GPs prescribing. RESULTS: GPs prescribed 45% and 51% of the prescriptions from physicians in 1988 and 1997, respectively, while the cost shares were 40% and 42%. An increase in prescriptions was seen both in relative and in absolute numbers (from 117414 in 3 months in 1988 to 161012 in 1995). The increase in cost per DDD (defined daily dose) during the study period was 47% for GPs and 72% for other doctors. CONCLUSIONS: GP prescribing increased in both absolute and relative numbers, while the cost increase per DDD was moderate compared to other physicians. PMID- 12564577 TI - Do general practitioners use what's in their doctor's bag? AB - OBJECTIVE: To find out what general practitioners (GPs) take with them on house calls and how frequently they use it? DESIGN: A questionnaire about the drugs and medical equipment used during house calls. SETTING: Two peer review groups of GPs in Overijse and Hoeilaart (Belgium). SUBJECTS: 29 GPs. MEAN OUTCOME MEASURES: Availability and use of drugs and medical equipment on house calls. RESULTS: All GPs had a stethoscope, a sphygmomanometer, an otoscope and sterile injection syringes at their disposal on house calls and they used them frequently. Only 57% took a blood glucose sensor with them and 25% took the medical records on home visits. Though only 50% always carried all of the most common emergency drugs with them, almost 100% of GPs had administered all of them in the previous 12 months. Only epinephrine and atropine were not frequently administered. CONCLUSION: Most of the GPs were sufficiently equipped to meet most situations that can occur during house visits and emergency calls. Most of the available drugs and equipment were used during the 12 months preceding the registration. Only the medical records and a blood glucose sensor were insufficiently available. PMID- 12564578 TI - Mutual understanding: a communication model for general practice. AB - OBJECTIVE: To present our pursuits towards development of a simple model for clinical communication intended for application by the practitioner as a tool for enhancing mutual understanding. DESIGN: Inspired by theories about patient centredness and interactive modes of understanding, and supported by the perspectives of the Danish philosopher Niels Thomassen, we reviewed audiotapes from our own consultations. Recognising four dimensions assumed to be essential for mutual understanding in the transcripts, we explored these dimensions further. RESULTS: We present a communication model consisting of the following dimensions: The Framework, within which the communication takes place; The Subject, about which the communication takes place; The Persons, between whom the communication takes place; and The Action, verbally and non-verbally, through which communication takes place. We describe these dimensions in detail. The nature of the dimensions indicates that there is an interrelationship between them, implying that the character of the communication may change if one of the factors is changed. CONCLUSION: Analysis of an ongoing or recent consultation completed in accordance with these four dimensions allows the doctor to refocus the communication, thus leading to a more extensive mutual understanding and perhaps enhanced freedom of action. PMID- 12564579 TI - The concept of risk: a comment to Reventlow and co-workers. PMID- 12564580 TI - A full plate. PMID- 12564581 TI - NAS reports on potential risks of animal biotechnology. PMID- 12564582 TI - What are the steps of the Educational Commission for Foreign Veterinary Graduates Certification Program. PMID- 12564583 TI - Vaccination information reaches audience. PMID- 12564584 TI - Farm animal production commentary ruffles feathers. PMID- 12564585 TI - Farm animal production commentary ruffles feathers. PMID- 12564586 TI - More attention warranted on Brucella abortus in animals. PMID- 12564587 TI - Scientific and moral considerations for live market practices. PMID- 12564588 TI - What is your diagnosis? Cholestasis, hepatic cholelithiasis. PMID- 12564589 TI - General toxicologic hazards and risks for search-and-rescue dogs responding to urban disasters. AB - In large-scale disasters, it is not always possible to identify every potential toxic agent to which SAR dogs may be exposed. However, an understanding of the basic means by which dogs may be exposed to toxic agents can aid veterinarians in determining basic risks for particular SAR sites and allow veterinarians to institute general preventive measures (eg, frequent eye washes) to minimize exposure. Discussions with public health and other authorities on-site may aid in identifying site-specific risks for SAR dogs. Finally, ensuring that SAR dog handlers are aware of basic risks, precautions, and decontamination measures is essential, as handlers are the first line of defense in preventing illness or injury to SAR dogs as they work a disaster area. PMID- 12564590 TI - Toxicologic agents of concern for search-and-rescue dogs responding to urban disasters. PMID- 12564591 TI - Management and prevention of toxicoses in search-and-rescue dogs responding to urban disasters. PMID- 12564592 TI - Employment, starting salaries, and educational indebtedness of year-2002 graduates of US veterinary medical colleges. PMID- 12564593 TI - Evaluation of day-to-day variability of serial blood glucose concentration curves in diabetic dogs. AB - OBJECTIVE: To evaluate day-to-day variability of serial blood glucose concentration curves in dogs with diabetes mellitus. DESIGN: Prospective clinical study. ANIMALS: 10 dogs with diabetes mellitus. PROCEDURE: Paired 12-hour serial blood glucose concentration curves performed during 2 consecutive days were obtained on 3 occasions from each dog. Dogs received the same dose of insulin and meal every 12 hours on both days. For each pair of curves, comparison was made between the results of days 1 and 2. RESULTS: Mean absolute difference (without regard to sign) between days 1 and 2 for each parameter was significantly > 0, disproving the hypothesis that there is minimal day-to-day variability of serial blood glucose concentration curves when insulin dose and meals are kept constant. Coefficient of variation of the absolute difference between days 1 and 2 for each parameter ranged from 68 to 103%. Evaluation of the paired curves led to an opposite recommendation for adjustment of the insulin dose on day 2, compared with day 1, on 27% of occasions. Disparity between dosage recommendations was more pronounced when glucose concentration nadir was < 180 mg/dL (10 mmol/L) on 1 or both days. In this subset of 20 paired curves, an opposite recommendation for dosage adjustment was made on 40% of occasions. CONCLUSIONS AND CLINICAL RELEVANCE: There is large day-to-day variation in parameters of serial blood glucose concentration curves in diabetic dogs. Day-to-day variability of serial blood glucose concentration curves has important clinical implications, particularly in dogs with good glycemic control. PMID- 12564594 TI - Association between initial systolic blood pressure and risk of developing a uremic crisis or of dying in dogs with chronic renal failure. AB - OBJECTIVE: To determine whether high systolic blood pressure (SBP) at the time of initial diagnosis of chronic renal failure in dogs was associated with increased risk of uremic crisis, risk of dying, or rate of decline in renal function. DESIGN: Prospective cohort study. ANIMALS: 45 dogs with spontaneous chronic renal failure. PROCEDURE: Dogs were assigned to 1 of 3 groups on the basis of initial SBP (high, intermediate, low); Kaplan-Meier and Cox proportional hazards methods were used to estimate the association between SBP and development of a uremic crisis and death. The reciprocal of serum creatinine concentration was used as an estimate of renal function. RESULTS: Dogs in the high SBP group were more likely to develop a uremic crisis and to die than were dogs in the other groups, and the risks of developing a uremic crisis and of dying increased significantly as SBP increased. A greater decrease in renal function was observed in dogs in the high SBP group. Retinopathy and hypertensive encephalopathy were detected in 3 of 14 dogs with SBP > or = 180 mm Hg. Systolic blood pressure remained high in 10 of 11 dogs treated with antihypertensive drugs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that initial high SBP in dogs with chronic renal failure was associated with increased risk of developing a uremic crisis and of dying. Further studies are required to determine whether there is a cause-and-effect relationship between high SBP and progressive renal injury and to identify the risks and benefits of antihypertensive drug treatment. PMID- 12564595 TI - Postoperative hypoxemia and hypercarbia in healthy dogs undergoing routine ovariohysterectomy or castration and receiving butorphanol or hydromorphone for analgesia. AB - OBJECTIVE: To determine frequency and severity of postanesthetic hypoxemia and hypercarbia in healthy dogs undergoing elective ovariohysterectomy or castration and given butorphanol or hydromorphone for analgesia. DESIGN: Prospective trial. ANIMALS: 0 healthy dogs weighing > 10 kg (22 lb). PROCEDURE: Dogs were anesthestized with acepromazine, glycopyrrolate, thiopental, and isoflurane, and butorphanol (n = 10) or hydromorphone (10) was used for perioperative analgesia. Arterial blood gas analyses were performed 10 and 30 minutes and 1, 2, 3, and 4 hours after extubation. RESULTS: In dogs that received hydromorphone, mean PaCO2 was significantly higher, compared with the preoperative value, 10 and 30 minutes and 1, 2, and 3 hours after extubation. Mean PaCO2 was significantly higher in dogs given hydromorphone rather than butorphanol 10 and 30 minutes and 1 and 2 hours after extubation. Mean PaO2 was significantly lower, compared with preoperative values, 30 minutes and 1 and 2 hours after extubation in dogs given hydromorphone and 30 minutes after extubation in dogs given butorphanol. Mean PaO2 was significantly lower in dogs given hydromorphone rather than butorphanol 1 hour after extubation. Four dogs had PaO2 < 80 mm Hg 1 or more times after extubation. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of hydromorphone to healthy dogs undergoing elective ovariohysterectomy or castration may result in transient increases in PaCO2 postoperatively and that administration of hydromorphone or butorphanol may result in transient decreases in PaO2. However, increases in PaCO2 and decreases in PaO2 were mild, and mean PaCO2 and PaO2 remained within reference limits. PMID- 12564596 TI - Vitamin D-dependent rickets type 2 in a four-month-old cat. AB - Vitamin D-dependent rickets type 2 in a four-month-old cat A 4-month-old male domestic shorthair cat was examined because of lethargy, vomiting, diarrhea, muscle tremors, and mydriasis. Laboratory evaluation revealed hypocalcemia, hyperphosphatemia, and high intact parathormone and calcitriol concentrations. Findings were compatible with a diagnosis of vitamin D-dependent rickets type 2. Treatment consisted of oral administration of calcium and calcitriol supplements. During the subsequent 18 months, the cat remained clinically normal. Treatment with oral calcium supplements was eventually discontinued, and the cat was able to maintain serum calcium concentrations within reference limits. PMID- 12564597 TI - Primary extraskeletal hepatic osteosarcoma in a cat. AB - A 13-year-old spayed female domestic shorthair cat with an abdominal mass was evaluated; radiography revealed a radiopaque mass in the cranioventral region of the abdomen. A celiotomy was performed, and the mass was identified histologically as a hepatic osteosarcoma. Complete remission of the tumor was accomplished after partial hepatectomy and adjuvant treatment with carboplatin. PMID- 12564598 TI - Use of a hydroxyapatite orbital implant in a cosmetic corneoscleral prosthesis after enucleation in a horse. AB - A hydroxyapatite orbital implant was used after enucleation of an eye from a 5 year-old performance horse. A custom-made corneoscleral prosthesis was made and fitted over the hydroxyapatite implant. The implant and surgery were well tolerated. Placement of a cosmetic prosthesis is desired after enucleation of equine eyes to allow horses to return to competition. Synthetic spheres consisting of methylmethacrylate or silicone have been used, although reported complications have included extrusion, infection, and poor cosmetic results. Hydroxyapatite orbital implants made from marine coral allow vascular and fibrous tissue growth from the host orbit into the implant, which decreases the possibility of implant extrusion or infection and allows enhanced healing of defects in the overlying conjunctiva. Extraocular muscle fixation onto the implant permits motility and assists in the prevention of implant extrusion. PMID- 12564599 TI - Intraosseous regional perfusion for treatment of septic physitis in a two-week old foal. AB - A 2-week-old Morgan filly examined because of lameness of 5 days' duration was found, on the basis of clinical and radiographic findings, to have septic physitis of the distal end of the radius. The foal was treated by means of intraosseous regional perfusion with penicillin and amikacin and systemic administration of antimicrobials. Intraosseous regional perfusion was performed 3 times. The foal was anesthetized for the first episode of intraosseous regional perfusion, but was only sedated for the subsequent 2 episodes. Antimicrobials were administered systemically for 22 days. Foals with septic physitis are typically considered to have a guarded to poor prognosis for recovery; however, this treatment regimen resulted in relatively fast and uncomplicated resolution of the infection in this foal. PMID- 12564600 TI - Analysis and testing of biological stains--the Biological Stain Commission Procedures. PMID- 12564601 TI - A differential staining method to identify lignified and unlignified tissues. AB - We investigated the use of safranin O and astra blue dissolved in ethyl alcohol as differential stains to distinguish between lignified and unlignified tissues in microtome sections of tension and normal wood of sugar and red maple. Normal wood was used as a control for the histochemical analysis. Lignified and unlignified tissues were found in the same section for both tension and normal wood of each species. These results were confirmed in unstained samples using ultraviolet light. Unlignified libriform fibers were detected using both techniques. Libriform fibers did not fluoresce in UV light, although fluorescence was observed in some of the cell corners. The astra blue in ethyl alcohol and the UV wavelength we used differentiated syringyl from guaiacyl lignins. Ethyl alcohol solutions of these dyes provide an effective and reliable method to distinguish lignified and unlignified tissues. PMID- 12564602 TI - An automated rotisserie system for processing Western blots. AB - An apparatus to automate completely the processing of Western blots is described. The prototype is based on a popular rotisserie system design. The incubation chamber consists of an inner cylinder that rotates inside an outer cylinder (incubation chamber). The blot is contained in the inner cylinder. Two magnets are mounted at one end of the inner cylinder, and rotation of the inner cylinder is effected by two magnets mounted on a motor drive outside the incubation chamber. Movement of chemicals into and out of the incubation chamber is driven pneumatically, and the entire process is controlled by a computer. Processing a blot for chemiluminescent detection takes 7 h to complete without human intervention. The quality of the resulting image is comparable to or better than a blot using manual processing. In addition, the prototype is capable of re collecting all three antisera for future use. PMID- 12564603 TI - Selective fluorescence of zymogen granules of pancreatic acinar cells stained with hematoxylin and eosin. AB - Following staining with hematoxylin and eosin Y, paraffin sections of mouse pancreas were examined by transmitted light, epifluorescence and confocal laser scanning microscopy. Light microscopy revealed that the nuclei of pancreatic acinar cells were located basally, while the apices of the cells appeared eosinophilic, although the secretory granules were difficult to visualize. Under violet-blue light excitation, the zymogen granules at the apices of the acinar cells showed strong yellowish fluorescence; the other part of the cytoplasm was only faintly fluorescent and the nuclei and the supporting tissues were nonfluorescent. Confocal laser scanning microscopy resulted in clear pictures of the zymogen granules and their distribution within the cell. The fluorescent emission of zymogen granules was certainly the result of eosin Y staining, because hematoxylin is not a fluorochrome and the zymogen granules are not autofluorescent. Staining with eosin Y alone, however, did not result in clear fluorescent images of zymogen granules or any other cellular structures. Our observation shows that the fluorescence emission of eosin Y allows easy and precise recognition of zymogen granules of pancreatic cells. PMID- 12564604 TI - Ralph D. Lillie: eminent histochemist and pathologist. PMID- 12564605 TI - Introduction. The symposium that never occurred: pre-clinical and clinical development of sibenadet. PMID- 12564606 TI - Dual dopamine D2 receptor and beta2-adrenoceptor agonists for the treatment of chronic obstructive pulmonary disease: the pre-clinical rationale. AB - This paper describes the rationale for the development of dual dopamine D2 receptor and beta2-adrenoceptor agonists as potential treatments for the symptoms of chronic obstructive pulmonary disease (COPD). The putative involvement of pulmonary sensory afferent nerves in mediating the key COPD symptoms of breathlessness, cough and excess sputum production is outlined and the hypothesis that activation of D2-receptors on such nerves would modulate their activity is developed. This premise was tested, in a range of animal models, using the first of a novel class of dual dopamine D2-receptor and beta2-adrenoceptor agonists, sibenadet HCl (Viozan, AR-C68397AA). In the course of these studies it was demonstrated that sibenadet, through activation of D2-receptors, inhibited discharge of rapidly adapting receptors and was effective in reducing reflex induced tachypnoea, mucus production and cough in the dog. Sibenadet, through its activation of beta2-adrenoceptors, was also shown to be an effective bronchodilator with a prolonged duration of action following topical administration to the lungs. These studies also indicated that sibenadet had a wide therapeutic ratio with respect to expected undesirable side-effects such as emesis and cardiovascular disturbances. These results provided a compelling rationale for the initiation of a clinical development programme with sibenadet for the treatment of COPD. PMID- 12564608 TI - The role of the novel D2/beta2-agonist, Viozan (sibenadet HCl), in the treatment of symptoms of chronic obstructive pulmonary disease: results of a large-scale clinical investigation. AB - Viozan (sibenadet HCl, AR-C68397AA) is a novel dual D2 dopamine receptor, beta2 adrenoceptor agonist, developed specifically to treat the key symptoms of chronic obstructive pulmonary disease (COPD), breathlessness, cough and sputum. The dual sensory nerve modulation and bronchodilator effects of sibenadet have been demonstrated in initial dose-ranging studies of patients with COPD and large scale clinical evaluation has now been completed. Sibenadet efficacy was determined by assessing symptomatic changes, as defined by the novel assessment tool, the Breathlessness, Cough and Sputum Scale (BCSS). The findings of two placebo-controlled studies are reported. These multicentre, double-blind, placebo controlled studies recruited over 2000 patients with stable COPD, randomized to receive sibenadet (500 microg) or placebo, pressurized metered-dose inhaler (pMDI) (three times daily) for a period of 12 or 26 weeks. Diary cards were completed daily by patients throughout the study to record BCSS scores, peak expiratory flow (PEF), study drug and rescue bronchodilator usage, changes in concomitant medication and adverse events. The primary endpoints were defined as change from baseline to the final 4 weeks of the treatment period in mean BCSS total score, and forced expiratory volume in one second (FEV1) measured 1 hour after administration of the final dose of study drug and expressed as a percentage of the predicted FEV1. In addition, clinic assessments were made to determine changes in pulmonary function, health-related quality of life, perception of treatment efficacy and adverse events. Despite initial improvements in mean daily BCSS total scores in patients receiving sibenadet, the difference in the change from baseline to the final 4 weeks of the treatment period between the two treatment groups was neither statistically significant, nor considered to be of clinical importance. Although marked bronchodilator activity was seen early on with sibenadet treatment, the duration of effect diminished as the studies progressed. Sibenadet use was not associated with any safety concerns. These studies, utilizing the novel BCSS, have clearly illustrated that, despite initial symptomatic improvement with sibenadet therapy, this clinical benefit was not sustained over the course of the study. PMID- 12564607 TI - Early clinical investigation of Viozan (sibenadet HCl), a novel D2 dopamine receptor, beta2-adrenoceptor agonist for the treatment of chronic obstructive pulmonary disease symptoms. AB - Viozan, (Sibenadet HCl, AR-C68397AA) is a dual D2 dopamine receptor, beta2 adrenoceptor agonist that combines bronchodilator activity with the sensory afferent modulating effects associated with D2-receptor agonism. Investigation in animal models of key chronic obstructive pulmonary disease (COPD) symptoms has demonstrated that sibenadet effectively inhibits sensory nerve activity, thereby reducing reflex cough, mucus production and tachypnoea. The results of the early clinical evaluation of this novel agent are reported. An initial proof of concept study (Study 1) aimed to determine the clinical potential of this novel agent by assessing the effects of three doses of sibenadet therapy relative to placebo, with two commonly used bronchodilators, intended to provide a benchmark against which sibenadet activity could be judged. In all, 701 patients were randomized to one of three sibenadet dose groups (400, 600 or 1000 microg ex valve), salbutamol 200 microg, ipratropium bromide (IB) 40 microg or placebo, all three times daily via pressurized metered dose inhaler (pMDI) for 4 weeks. Once the results of Study 1 had been evaluated, a dose-ranging, study (Study 2) involving 872 patients randomized to receive sibenadet (45, 270, or 495 microg ex actuator), or placebo all three times daily via pMDI, for 6 weeks commenced. Both studies were preceded by a 2-week baseline phase and followed by a 2-week follow up period.The primary efficacy variable identified changes in key COPD symptoms over the treatment period (compared with baseline data) as determined by the novel Breathlessness, Cough and Sputum Scale (BCSS). In addition, data on lung function, health-related quality of life and adverse events were collected. Patients receiving sibenadet therapy three times daily exhibited statistically significantly greater improvements in BCSS total score than those receiving placebo or bronchodilator therapy alone. A clear dose-response was evident in Study 2. Symptom improvement in this study was also accompanied by improved lung function and health-related quality of life. Sibenadet therapy was well tolerated with an adverse events profile comparable to current bronchodilator therapy. These data were viewed as extremely encouraging, warranting further, large-scale clinical investigation. PMID- 12564609 TI - Symptoms are an important outcome in chronic obstructive pulmonary disease clinical trials: results of a 3-month comparative study using the Breathlessness, Cough and Sputum Scale (BCSS). AB - The need to manage the key symptoms of chronic obstructive pulmonary disease (COPD) (breathlessness, cough and sputum) is an important treatment objective. Viozan (sibenadet HCl, AR-C68397AA) is a novel dual D2 dopamine receptor, beta2 adrenoceptor agonist, which combines conventional bronchodilatory activity with the sensory nerve modulation afforded by dopamine agonism. The efficacy of this agent in relieving patient symptoms has been determined in a series of large scale clinical studies; the results of a 3-month, placebo-controlled multi-centre study are reported. Effect on patient symptoms was determined using a novel patient-reported assessment instrument, the Breathlessness, Cough and Sputum Scale (BCSS). Patients with smoking-related COPD were required to complete a 2 week baseline period before being randomized to one of three treatment groups; sibenadet (500 microg three times daily) plus placebo (twice daily); salmeterol (50 microg twice daily) plus placebo (three times daily); placebo (twice daily) plus a second placebo (three times daily). All treatments were delivered via pressurized metered dose inhaler (pMDI) for 12 weeks. From enrolment, patients were required to complete daily diary cards to record symptoms of breathlessness, cough and sputum, medication use and adverse events. The primary outcome measure was the difference between the mean BCSS total score measured over the baseline period and the mean BCSS total score in the final 4 weeks of the treatment period. Secondary measures included assessment of lung function, rescue medication use, exacerbations, health-related quality of life, opinion of efficacy and safety. Although an initial reduction in BCSS total score (indicating symptom improvement) was seen with sibenadet therapy, this effect was not maintained for the study duration. Salmeterol therapy, however, resulted in a sustained reduction in BCSS total score. No notable benefit over placebo was seen in lung function, exacerbations or health-related quality of life with either active treatment. While the results of this study failed to demonstrate sustained efficacy with sibenadet therapy, they do indicate the value of symptom assessment in the clinical evaluation of new drugs for the treatment of COPD. PMID- 12564610 TI - Long-term use of Viozan (sibenadet HCl) in patients with chronic obstructive pulmonary disease: results of a 1-year study. AB - Viozan (sibenadet HCl, AR-C68397AA) is a novel dual D2 dopamine receptor, beta2 adrenoceptor agonist that has been investigated for efficacy in alleviating the symptoms of chronic obstructive pulmonary disease (COPD). The slowly progressive nature of this disease means that patients will require ongoing therapeutic management for many years, or even decades. With such long-term treatment, the safety profile of new agents will be of paramount importance. As part of the large-scale assessment of sibenadet, a 12-month safety study has been conducted. Following completion of a 2-week baseline period, 435 adults with stable, symptomatic, smoking-related COPD were randomized to receive either 500 microg sibenadet or placebo delivered via pressurized metered dose inhaler (pMDI), three times daily for 52 weeks. Sibenadet therapy was generally well tolerated, with the only notable differences seen in the incidence of tremor and taste of treatment (16.9% vs. 4.1% and 14.5% vs. 4.1% in the sibenadet and placebo groups respectively). There were a total of 79 patients with serious adverse events (SAEs), 43 (14.8%) in the sibenadet pMDI group and 36 (24.8%) in the placebo group. No clinically significant abnormal laboratory values or overall differences between treatment groups were noted. Similarly, there were no clinically significant differences between the two treatment groups for cardiac variables, or in vital signs. The secondary variables showed no notable differences with respect to lung function, exacerbations or health-related quality of life. Due to the effective beta2-agonist properties, patients in the sibenadet group did, however, report reduced rescue medication usage at all timepoints. While the results of this study show that, overall, sibenadet therapy was well tolerated, the lack of sustained benefit reported in large-scale clinical efficacy studies means that sibenadet development will not be continued. PMID- 12564611 TI - Utilization of health care services by patients with chronic obstructive pulmonary disease. AB - In order to identify healthcare resource use patterns associated with chronic obstructive pulmonary disease (COPD), resource utilization (RU) data collection was integrated into a randomized, double-blind placebo-controlled study of Viozan (sibenadet HCl). This study enrolled patients with symptomatic, smoking-related COPD, randomized to receive sibenadet or placebo for a 52-week treatment period. A questionnaire establishing typical pre-trial, COPD-related RU was completed by each patient. Subsequent data were collected by means of an Interactive Voice Response System (IVRS) at 30-day intervals (14 time points) during the study and in the follow-up period. The IVRS system facilitated data collection and minimized inconvenience to the patient. Compliance with the requirement to record details of the healthcare services during the year-long study was high. No overall trend for lower RU was associated with sibenadet therapy, which correlates with the lack of sustained clinical effect seen in studies conducted concurrently. These data do, however, provide valuable information on RU associated with COPD and insights into adjustments associated with changes in disease course. Physicians were seen to be the most common source of care for patients with COPD and more of the patients with severe COPD (stage III) than mild (stage I) were seen to utilize the most expensive resources (e.g. inpatient hospital care). For those patients who experienced an exacerbation during the trial (irrespective of treatment group), resource use was increased during the periods when an exacerbation was reported when compared with the periods before or after an exacerbation. The proportion of cases attending the physician doubled and with a trip to the Emergency Room (ER) increased approximately ninefold during the reporting period in which the exacerbation occurred compared with the previous month. This study has shown that use of an IVRS, even in elderly patients, is an effective means of gathering RU data over long periods. The study findings suggest that the advent of effective therapeutic interventions, particularly any with the ability to minimize exacerbations and limit disease progression, could impact on the health care services used and potentially reduce associated costs. PMID- 12564612 TI - Evaluating symptoms in chronic obstructive pulmonary disease: validation of the Breathlessness, Cough and Sputum Scale. AB - This study examined the reliability and validity of a new measure for evaluating symptoms in patients with chronic obstructive pulmonary disease (COPD): the Breathlessness, Cough and Sputum Scale (BCSS). Designed as a daily diary, the BCSS is a patient-reported outcome measure that asks patients to rate the severity of the three symptoms, each on a 5-point scale; higher scores indicate more severe symptoms. Item scores are summed to yield a total score. Secondary analysis of data were from two multinational trials (n = 629; 765). The BCSS item and total scores exhibited evidence of internal consistency (alpha = 0.70 daily; 0.95 to 0.99 over time) and reproducibility (ICC = 0.77 to 0.88). Correlations (r) with pulmonary function (FEV1 % predicted, PEF) were -0.01 (n.s.) to -0.36 (P < 0.001). Correlations with the St George's Respiratory Questionnaire total and SF-36 Physical Functioning subscale were 0.44 to 0.59 (P < 0.0001). Breathlessness and total scores differentiated patients by disease severity (P < 0.01) and rescue medication use (P < 0.01). Cough, sputum, and total scores increased with sputum volume (r = 0.27, 0.30, 0.31; P < 0.001). Patients for whom treatment was moderately or highly effective reported significant improvements in BCSS scores (P < 0.0001). Results suggest that the BCSS is a reliable, valid, and responsive patient-reported outcome measure of symptom severity in patients with COPD. PMID- 12564613 TI - Conclusion. Lessons from the novel D2 dopamine receptor, beta2-adrenoceptor agonist, Viozan: chronic obstructive pulmonary disease and drug development implications. AB - The development of novel drugs for the treatment of chronic obstructive pulmonary disease (COPD) poses significant challenges. The mechanisms through which the chronic symptoms of COPD arise are poorly understood, making identification of potential therapeutic targets and in vivo evaluation of potential therapies extremely difficult. Despite these challenges, a unique approach of combined D2 dopamine, beta2-adrenoceptor agonism was identified as a valid potential target for the treatment of key COPD symptoms, the therapeutic potential of which was investigated in a series of preclinical evaluations. Subsequent clinical assessment has amassed a wealth of data from over 4000 patients, providing valuable insights into COPD, clinical trial design and the value of patient self assessment tools. PMID- 12564614 TI - Influence of blood sampling techniques on ionized magnesium level. AB - The aim of the study was to evaluate whether different blood sampling techniques (venous vs. capillary) influence the level of ionized magnesium. A total of 12 men and 20 women were included. A venous blood sample was collected from the antecubital vein without stasis in syringes containing 50 IU/ml electrolyte balanced heparin. Capillary tubes containing sodium heparin 50 IU/ml were used for blood sampling (150 microl) from the earlobe. A statistically significant difference between venous and capillary ionized magnesium at actual pH (0.57+/ 0.04 vs. 0.55+/-0.04 mmol/L, p<0.05) was observed, whereas no difference was seen between adjusted ionized magnesium. No difference was observed between venous and capillary ionized calcium at actual pH or adjusted to pH 7.4. The pH level was significantly higher in capillary blood (7.44+/-0.02 vs. 7.48+/-0.02. p<0.00001). The higher pH in capillary blood may be due to greater exposure to air in capillary tubes, and this most likely explains the observed difference between venous and capillary ionized magnesium at actual pH. PMID- 12564615 TI - Ambulatory blood pressure and left ventricular mass in older age. AB - BACKGROUND: The purpose of this study was to analyse the association of ambulatory blood pressure (ABP) to left ventricular mass (LVM) in a population aged over 64 years and to describe the level of ABP in subjects with and without left ventricular hypertrophy (LVH) in older age. METHODS: ABP measurement and echocardiography for calculation of LVM were assessed in 490 inhabitants (mean age 70.7 years, range 64-87 years) of a small town in southwestern Finland who were able to visit an outpatient clinic. Explanatory factors associated with LVM were assessed with linear regression analysis. LVH was defined as calculated LVM index values exceeding 150 g/m2 in men and 120 g/m2 in women adopted from the Framingham Study. RESULTS: Systolic ABP was significantly associated with LVM. No correlation between diastolic ABP and LVM was found. Other factors independently related to LVM were gender, body mass index and age. The prevalence of echocardiographic LVH was 22%. Subjects with LVH had markedly higher systolic ABP levels than those without LVH (mean (SD) 24-h ABP: 132(16)/75(8) mmHg vs. 123(13)/75(8) mmHg). CONCLUSION: Systolic ABP is associated with LVM in older people. In addition, systolic ABP is superior to diastolic ABP in relation to LVM in the aged. PMID- 12564616 TI - Novel serum inflammatory markers in acute appendicitis. AB - BACKGROUND: Cytokines and leukocyte adhesion molecules are activated and found in increased concentrations in bacterial infection. The purpose of this study was to investigate whether some of these new serum markers could be feasible as a single on-admission test to predict acute appendicitis (AA). METHODS: In an open prospective study the diagnostic potentials of two cytokine measurements (interleukin-6 and interleukin-8), soluble leukocyte adhesion molecule (CD44), C reactive protein (CRP) and white blood cell (WBC) count were compared in 80 consecutive patients who had undergone surgery for suspected AA. The diagnostic performance of each parameter was tested by using receiver operating characteristic (ROC) curves. RESULTS: Phlegmonous AA was found in 34%, gangrenous AA in 40% and perforated AA in 5% of the patients. The proportion of negative explorations was 21%. Preoperative serum concentrations of IL-6 and CRP were elevated only in gangrenous and perforated AA. The concentrations of IL-8 and CD44 remained unchanged in AA. The sensitivity (84%), specificity (79%) and diagnostic accuracy (82%) of IL-6 were higher than the values for CRP, WBC, IL-8 and CD44 in predicting AA. CONCLUSION: ROC analysis confirmed that IL-6 showed the best trend in the diagnosis of AA. However, the diagnosis of AA was not greatly improved by any of the new serum markers as single on-admission tests. PMID- 12564617 TI - Examination of the Metamax I and II oxygen analysers during exercise studies in the laboratory. AB - The performance of the Metamax I and the Metamax II portable analysers for measuring the O2 uptake has been examined during exercise. Healthy subjects ran on the treadmill or bicycled on ergometers while the O2 uptake was measured by the Metamaxes and also by the Douglas bag technique or the Vmax 29 instrument. In the first series of experiments, O2 uptake was measured by each instrument in turn. In later experiments two or more breathing valves were connected in a series, thus enabling measurement of the O2 uptake simultaneously by more than one instrument. The O2 uptake measured by the Metamax analysers rose linearly by the value given by the control methods. However, there were variations of approximately 5% because the relationships differed between subjects. When the data from each subject were examined separately, the error of regression was 0.5 1 micromol s(-1) kg(-1) (2-3%), and the error of regression when relating the O2 uptake to the exercise intensity was similar to that found when using the Douglas bag technique alone. In most cases the lung ventilation reported by the Metamaxes was a few percent less than that given by the control methods, while the fractional extraction of O2 was higher for the Metamaxes. The respiratory exchange ratios (R-value) reported by the Metamaxes were in good agreement with those of the control methods in the range 0.9-1.0 only; for this parameter, the Metamaxes do not seem to be reliable for exercise testing. The O2 uptake and the R-value were also calculated from the raw data reported by the Metamaxes. The calculated values differed somewhat from those reported by the instruments, and the calculated values were more in agreement with those obtained by the Douglas bag technique than those reported by the instrument. This study suggests that the O2 uptake reported by the Metamaxes is precisely measured within subjects but that there are some systematic errors as well as variations between subjects. PMID- 12564618 TI - Exhaustive exercise increases plasma/serum total oxidation resistance in moderately trained men and women, whereas their VLDL + LDL lipoprotein fraction is more susceptible to oxidation. AB - The purpose of this study was to evaluate the effects of exhaustive exercise (marathon run) on different lipid peroxidation measurements, including copper induced serum lipids and VLDL + LDL oxidation susceptibility, and on plasma total antioxidative capacity (TRAP), muscular damage and plasma antioxidants in healthy moderately trained male (n = 21) and female (n = 25) volunteers. Blood samples were taken before and just after the 42-km run. In women, baseline levels of several antioxidative compounds (serum albumin and uric acid, plasma free thiols and blood glutathione) were lower, resulting in 21.5% lower plasma total antioxidative capacity and 70.3% higher serum oxidation susceptibility, compared to men. To compare effects in men and women, the exercise-induced variable changes were adjusted for their baseline levels. After this adjustment, there were no statistically significant differences between the genders in the extent of muscular damage (serum creatine kinase, (CK)), or in the change in serum lipids or VLDL + LDL oxidation susceptibility, or that of plasma antioxidative capacity. A possible beneficial effect of exercise was that serum HDL cholesterol levels increased significantly in both genders, but especially in women. In the group of pooled genders (n=46), the increases in serum CK and in plasma lactate were 190% (95% CI, 133% to 246%) and 109% (95% CI, 65% to 175%), respectively. On the basis of our lipid peroxidation and TRAP measurements, uric acid was observed to be the most important plasma antioxidant. The effect of exercise was to decrease the oxidation susceptibility of serum lipids by 24.8% (95% CI 13.4% to 36.2%) and to elevate plasma TRAP by 14.6% (95% CI, 11.4% to 17.7%). Nonetheless, the oxidation susceptibility of the VLDL + LDL fraction increased by 11.0% (95% CI, 1.9% to 20.2%). Our results suggest that there are no gender-based differences in exhaustive exercise-induced lipid peroxidation or muscular damage. Secondly, even though exhaustive exercise can increase plasma/serum total resistance towards oxidation, the oxidation resistance of the atherogenic lipoprotein fraction might be diminished. On the basis of these results, several in vitro measurements of lipid peroxidation assessing both water and lipid soluble plasma fractions are needed if a true perspective of the plasma redox status is to be obtained. PMID- 12564619 TI - Upper reference limit, analytical quality specifications and clinical use of haemoglobin A1C. AB - Haemoglobin A1c (HbA1c) is now the key component for monitoring glycaemic control in diabetes mellitus (DM), especially for its close relation to diabetes complications. However, treatment goals in terms of HbA1c concentrations have been difficult to define and compare because of lack of international standardization and lack of common reference values of HbA1c concentrations. The aims of our study were to document our HbA1c analysis and make it traceable to international reference laboratories with the aid of current reference preparations, to establish a reference interval based on a low-risk population, and to evaluate the analytical quality specifications, which could meet clinical needs. The s(analytical) of our method (Tosoh) was < 0.3 HbA1c%, and the mean bias as estimated from Dr Cas Weykamp's reference preparation was below 0.3 HbA1c. This was the same as that for participating Scandinavian and international reference laboratories. The concentrations were made traceable to results from the Diabetes Control and Complication Trial (DCCT) and the UK Prospective Diabetes Study (UKPDS). Risk groups for DM were ruled out from a randomly selected population in Vejle County, which isolated a "low-risk" reference population. The 97.5 reference interval in this population (N=430) was from 5.07 HbA1c% (95% CI: 5.02-5.11) to 6.24 HbA1c% (95% CI: 6.19-6.30), and the 99.9 centile was 6.62 HbA1c% (95%) CI 6.55-6.71). Body mass index, age and gender contributed marginally to the level of HbA1c concentrations. A 10% delta risk estimate of DM complications was detectable with a probability of Type I error of 40%, while adoption of a significance level of 95% and consideration to biological variation needed a risk difference of at least 33% to be detected. The critical difference was 11% for changes in either direction at s(analytical) < or = 0.2 HbA1c% and a s(biological) of 0.3 HbA1c%. Based on criteria for sharing common reference intervals and clinical utility, we accepted that the bias and s(analytical) should both be < 0.3 HbA1c% at the level of 7.0 HbA1c%. PMID- 12564620 TI - Reproducibility of S-insulin and B-glucose responses in two identical oral glucose tolerance tests. AB - Recently, the diagnostic criteria for type 2 diabetes mellitus have been changed, but there are disagreements about which measurements should be used. In contrast to the American Diabetes Association (ADA), The World Health Organization (WHO) still recognizes fasting and 2-h glucose concentrations measured on either plasma or whole blood as diagnostic tools. Insulin sensitivity and insulin secretion are both assumed to be involved in the pathogenesis of type 2 diabetes. The oral glucose tolerance test (OGTT) for estimating insulin sensitivity and secretion is increasingly used, e.g. in intervention trials. The objectives of this study were to estimate the coefficients of intra-individual variation (CVw) of blood glucose and serum insulin concentrations from an OGTT as well as indices of insulin sensitivity (HOMA) and insulin secretion (delta insulin30/delta glucose30) derived from this test. Following duplicate OGTTs with a median interval of 13 days (range 1-87 days), the analytical, inter-individual, and intra-individual coefficients of variation were calculated by nested ANOVA. The CVw for fasting blood glucose (7%) was considerably lower than that for 2-h post-load glucose (15%), which was again lower than for the insulin concentrations and indices of insulin sensitivity and secretion. In conclusion, the intra-individual variation is larger for 2-h post-load glucose than for fasting glucose and may question the continued use of the 2-h post-load glucose value in the diagnosis of type 2 diabetes. PMID- 12564621 TI - Early predictors of school-age behavior problems and social skills in children with intraventricular hemorrhage (IVH) and/or extremely low birthweight (ELBW). AB - RESEARCH FINDINGS: Child and family characteristics from early infancy (birth-3 months adjusted age, AA) and toddler period (24-30 months) were examined as potential predictors of behavior problems and social skills at 7.5 years of age in 90 LBW infants with IVH and/or ELBW who were part of a longitudinal study of early intervention effectiveness. Severity of medical problems at birth and child temperament at 3 months AA were associated with future social skill levels, but not with behavior problems. Family variables in the first months of the child's life including low income, single parent household, and high parenting stress were significantly correlated with behavior problems at 7.5 years of age. Difficult temperament and elevated parenting stress in the toddler period were strongly associated with later behavior problems. Toddler developmental quotient (DQ) was a strong predictor of future social skills. PRACTICE OR POLICY: These data highlight the importance of early intervention that targets child behavior and emotional adjustment along with cognitive development, especially in low income and single parent households. Interventions that target parent-child interaction and parent stress are also crucial in these populations, even beyond the birth-three period. PMID- 12564622 TI - Atypical expressions of jealousy in infants of intrusive- and withdrawn-depressed mothers. AB - To determine whether atypical jealousy responses in infants of depressed mothers are differentiated by maternal nonoptimal interactive profiles, 12-month-old infants of intrusive-depressed and withdrawn-depressed mothers were observed with their mothers and a stranger in two contexts. In the first, infants were fully ignored in a less stressful context in which the adults focused on a picture book. In the second, infants were fully ignored in a more stressful, jealousy inducement context, in which the adults attended to an infant-like doll. Cross context comparisons revealed that the jealousy-inducement condition was associated with infants of intrusive-depressed mothers demonstrating greater play, and lesser proximal and distal behaviors toward their mothers. In contrast, the jealousy-inducement condition was associated with infants of withdrawn depressed mothers directing greater proximal contacts toward the stranger. This investigation identified that as early as infancy, dysregulated expressions of jealousy are differentiated by depressed mothers' nonoptimal interactive patterns. Findings of this preliminary study call for investigative attention to this uncharted area of inquiry into mental health, and specifically, works addressing the importance of maternal characteristics to the unfolding of normative and atypical jealousy in infancy. PMID- 12564623 TI - Self-reported depressive symptoms in a school sample of Italian children and adolescents. AB - This study reports on self-reported depressive symptoms, assessed with the Children's Depression Inventory (CDI), in a school sample of 685 Italian students. The participants were 254 males and 431 females, the age range of the subjects was 8 to 17 years (mean age 13.0 +/- 2.8 years). The CDI mean score was 10.49 +/- 2.76. According to our data, about 10% of the subjects scored higher than the clinical threshold of 20. More than 1/3 of the students reported thoughts of death, but only 2.2% had explicit suicidal ideation. Girls scored higher than boys, and young adolescents (11-13 years) scored lower than pre pubertal children and older adolescents, and showed fewer thoughts of death. Factorial analysis yielded seven factors, a general factor with no predominant theme, two factors related to academic self-image and body image, and four related to anhedonia-withdrawal, hypocondria-asthenia, irritability-opposition, and loss of appetite. PMID- 12564624 TI - The implementation of families first in The Netherlands: a one year follow-up. AB - This study examined whether the American family preservation program Families First was successfully implemented in The Netherlands. Data were collected on 250 children of 177 families who received Families First. At the start of treatment 78% of the children appeared to have serious behavioral problems, 67% of the parents experienced a high level of parental stress, and 63% of the children went through a substantial number of life events during the year preceding the treatment. On average the treatments had the intended duration (about 4 weeks), intensity (about 10 hours a week) and availability (during working hours as well as in evenings and in weekends), and family workers did adhere to important guidelines of treatment delivery. One year after treatment 76% of the children were still living at home. Moreover, children's behavioral problems, parental stress and the number of life events turned out to be significantly decreased. It was concluded that Families First had reached its intended target group, delivered the treatment as intended, and achieved its intended outcomes, suggesting a successful implementation in The Netherlands. PMID- 12564625 TI - Temperament in adolescents with anxiety and depressive disorders and in their families. AB - Aim of this study was to investigate whether specific temperamental features were associated with anxiety and depressive disorders in adolescents, in their siblings and in their parents. Thirty adolescents with Anxiety disorders and 25 with both Anxiety and Depressive disorders were compared to 25 adolescents with learning disorders and to 28 normal subjects. Temperament in subjects and relatives was assessed by their parents with the EAS questionnaire. Subjects with Anxiety and Anxiety-Depression and their siblings showed higher scores on Emotionality and Shyness than Learning Disability and Normal subjects. Mothers and fathers of subjects from the Anxiety-Depression group had the highest Emotionality score. These findings suggest that both Emotionality and Shyness are prominent temperamental features in adolescents with anxiety with or without depression, and in their parents and siblings. PMID- 12564626 TI - Fluorescence in situ hybridisation analysis and ovarian histology of women with Turner syndrome presenting with Y-chromosomal material: a correlation between oral epithelial cells, lymphocytes and ovarian tissue. AB - The early detection of Y-chromosomal material in women with Turner syndrome (TS) is of great importance due to a relatively high risk of gonadal tumour development. Using fluorescence in situ hybridisation (FISH) analysis, we studied the presence of three different Y-specific sequences (SRY, Ycen and Yq12) in three different tissues (oral epithelial cells, lymphocytes and ovarian tissue) of twelve TS women. We have also described their ovarian histology. Two of the women (17%) had gonadal tumours. In five women where ovarian tissue was available, the presence of Y-chromosomal material in oral epithelial cells and lymphocytes correlated to the presence of Y-chromosomal material in the gonads. We therefore conclude that FISH analysis of oral epithelial cells and/or lymphocytes is a valuable complement to karyotyping for the early detection of Y chromosomal material in TS women. PMID- 12564627 TI - Genetic variation in three species of Hordeum, and the selection of accessions for the Barley Core Collection. AB - Starch gel electrophoresis of isozymes was used to investigate the level and distribution of genetic variation in accessions of three wild, diploid Hordeum species. Allelic variation was assessed for seven isozyme loci and used in the statistical analysis of 18 accessions of Hordeum brachyantherum subsp. californicum, 29 accessions of H. murinum subsp. glaucum and 11 accessions of H. pusillum. A major purpose with this study was to evaluate the selection of accessions for the international Barley Core Collection, complementing existing passport data. Allelic variation was found in all three species. The variation was mainly between accessions, as most accessions were fixed for certain alleles. Since all three species are self-pollinated, this was anticipated. The fact that the accessions have undergone one or several regeneration cycles from a rather limited number of individuals, after being collected, likely also have reduced the variation. The accessions were clustered by subjecting the genetic distances between them to UPGMA statistical treatment. The clusters in the dendrograms displayed for H. brachyantherum subsp. californicum and H. pusillum distinct patterns related to the geographical origin of the accessions. The accessions of H. murinum subsp. glaucum were less variable than the other two, and did not reveal such a clear pattern. By comparing the already made, geographically based, selection of accessions with the results from this study, it was evident that the selection had worked fairly well for H. brachyantherum subsp. californicum, but less so for the other two species. PMID- 12564628 TI - Physical mapping of the 5S and 45S rDNA in teosintes. AB - The physical locations of the 5S and 45S rDNA sequences were examined in three types of teosinte, Zea mays ssp. mexicana (2n = 20), Zea diploperennis (2n = 20) and Zea perennis (2n = 40) by biotinylated fluorescence in situ hybridization (FISH). The tested materials only showed one hybridization site of 5S rDNA on their genomes, but they were different in the position of the signals. The hybridization site of Zea mays ssp. mexicana was located on the long arm of chromosome 2, indicating that it is the same as the cultivated maize in the position of 5S rDNA, while the sites of Zea diploperennis and Zea perennis were on the short arms of other chromosomes. For 45S rDNA, one hybridization site was detected at secondary constriction region of the satellite chromosomes in Zea mays ssp. mexicana and Zea diploperennis, while in Zea perennis, besides the site located at the secondary constriction region, a second site on the short arm of another chromosome pair was observed. Our results provide additional evidence for Zea mays ssp. mexicana being a subspecies of Zea mays. PMID- 12564629 TI - Optimization of sorghum transformation parameters using genes for green fluorescent protein and beta-glucuronidase as visual markers. AB - Early and reliable detection of plant transformation events is essential for establishing efficient transformation protocols. We have compared the effectiveness of using the gene encoding a green fluorescent protein (GFP) and a beta-glucuronidase (gus) as reporter genes for early detection of transgene expression in explants subjected to biolistic bombardment and Agrobacterium mediated transformation. The results indicate that gfp gene is superior to gus gene in following transgene expression in transiently transformed materials in both methods of transformation. Using GFP as the screenable marker, we have optimized sorghum transformation with respect to the conditions for transformation, type of explants, promoters, and inbreds. These optimized conditions have been used to obtain stably transformed explants for subsequent regeneration. PMID- 12564630 TI - Edible oil crops and their integration with the major cereals in North Shewa and South Welo, Central Highlands of Ethiopia: an ethnobotanical perspective. AB - A total of 1050 sorghum (Sorghum bicolor) and tef (Eragrostis tef) fields distributed in six study sites within north Shewa and south Welo (Central Highlands of Ethiopia) were systematically surveyed to examine the status of integration of edible oil crops into the cereal-based farming system. Farmers' criteria for, and perception on, integration between edible oil crops and the major cereals at field level as well as integration between the cereal grains and the oilseeds in food making at home were studied and analyzed based on formal semi-structured interview and informal discussion with local expert farmers as key informants. Farmers' traditional space optimization technique has been instrumental in rightly fitting edible oil crops (as intercrops and border crops) into the cereal-based system. Six species of edible oil crops are integrated (70.3%) in various proportions in fields of sorghum and tef. At least one oil crop was significantly intercropped and/or border cropped in sorghum fields. Noog (Guizotia abyssinica) and sesame (Sesamum indicum) were the most important edible oil crops of the study area having strong integration with sorghum both at field and home level. On average, noog was more frequently intercropped with sorghum (8.3%) than with tef (4.5%), while it was more frequently border cropped with tef (32.4%) than with sorghum (19%). Sorghum was more frequently inter/border cropped with sesame (39/2%) than with tef (10.1/0.5%). The stronger the integration of a given oilseed with sorghum-based foods, the higher the companionship between sorghum fields and the oil crop in the landscape. This cultural practice by farmers has positive contributions to on-farm conservation of oil crops along with tef and specific sorghum landraces. The central theme of this paper therefore converges on the issue of on-farm in-situ agrobiodiversity conservation that was shaped by successive ancestral generations and passed on to the present. PMID- 12564631 TI - The control of gene conversion properties and corresponding-site interference: the effects of conversion control factor 5 on conversion at locus w9 in Ascobolus immersus. AB - The controls of various aspects and parameters of gene conversion at locus w-9 in the fungus Ascobolus immersus were investigated, along with positive and negative corresponding-site interference in meiotic chromatid pairing. When conversion control factor 5 alleles A and B were heterozygous, conversion frequencies at the closely linked target locus w9 were reduced to 3%, compared with 10.7% when A or B was homozygous, through effects on hybrid-DNA (h-DNA) formation parameters gamma1 and gamma2. In different ways, not related to whether ccf-5 alleles were homozygous or heterozygous, ccf-5 also affected parameters relating to the relative frequencies of asymmetric and symmetric h-DNA, the frequency with which the chromatid bearing the wild-type allele was the invading chromatid in asymmetric h-DNA, and h-DNA correction parameters for the frequency and direction of correction of mispairs. Corresponding-site interference is interference between the two pairs of non-sister chromatids of a bivalent in h-DNA formation at exactly corresponding sites. This interference was positive in the high conversion frequency crosses homozygous for ccf-5 alleles but was strongly negative in the low conversion frequency crosses heterozygous for ccf-5 alleles, through differential effects on parameters gamma1 and gamma2. Models of chromatid pairing are discussed. PMID- 12564632 TI - Genetic basis of plant height and its degree of indetermination in mungbean (Vigna radiata (L.) Wilczek). AB - The genetic basis of plant height at various growth stages and the degree of indetermination of plant height in six mungbean genotypes (NM 92, 6601, NM 89, VC 1560D and VC 3902A) were assessed through half diallel cross. Cultivars, 6601 and NM 92, were the best general combiner for pre-flowering dry matter accumulation and minimum increase in plant height from first flower to 90% pods maturity, respectively. For these traits, the combination NM 92 x NM 89 was the best specific combiner of all the crosses. Both additive and dominant gene effects controlled the inheritance of plant height at first pod and to 90% pods maturity, degree of indetermination of plant height (DDh) from first flower to first pod maturity (DDh1), DDh from first flower to 90% pods maturity (DDh2) and DDh from first pod maturity to 90% pods maturity (DDh3). Plant height at first flower was additively inherited. The additive gene action was predominant as compared to dominant gene action for all the traits examined. High narrow and broad sense heritability estimates for DDh2 showed that better response to selection is possible for the development of mungbean genotypes with minimum increase in plant height during post-flowering development. PMID- 12564633 TI - Chromosome analysis in 31 cases of benign and malignant breast tumors: a study in Brazil. AB - Cultures of 31 breast tumors, being 20 carcinomas and 11 benign lesions, were cytogenetically analysed. Clonal chromosome aberrations were detected in 16 carcinomas and in 4 benign lesions. Nine carcinomas and 2 benign lesions had multiple cytogenetically unrelated and related clones, whereas a single abnormal clone was observed in 7 carcinomas and in 2 benign lesions. Polyploid clones were found in 7 carcinomas and in 2 benign lesions. The presence of clonal chromosome aberrations and polyploid cells was not associated with the clinicopathologic parameters tested. Carcinomas had more clonal changes than benign lesions (p = 0.031), showing that cytogenetic features are of diagnostic value and that different chromosome anomalies might have different pathogenetic and prognostic significance. PMID- 12564634 TI - Repeated DNA sequences in the microbat species Miniopterus schreibersi (Vespertilionidae; Chiroptera). AB - Repetitive DNA sequences represent a substantial component of eukaryotic genomes. These sequences have been described and characterized in many mammalian species. However, little information about repetitive DNA sequences is available in bat species. Here we describe an EcoRI family of repetitive DNA sequences present in the species Miniopterus schreibersi. These repetitive sequences are 57.85%, A-T rich, organized in tandem, and with a monomer unit length of 904 bp. Methylation analysis using the isoesquizomer pair MspI and HpaII indicates that the cytosines present in the sequences CCGG are partially methylated. Furthermore, Southern blot analysis demonstrated that these DNA sequences are absent in the genomes of four related microbat species and suggest that it could be specific to the M. schreibersi genome. PMID- 12564635 TI - Primers for amplifying approximately 820 bp of highly polymorphic mitochondrial COI gene of Gyrodactylus salaris. PMID- 12564636 TI - G/Q-banded chromosome nomenclature of the Chinese raccoon dog, Nyctereutes procyonoides procyonoides Gray. PMID- 12564637 TI - Cardiovascular effects of 1,8-cineole, a terpenoid oxide present in many plant essential oils, in normotensive rats. AB - The cardiovascular effects of i.v. treatment with 1,8-cineole, a monoterpenic oxide present in many plant essential oils, were investigated in normotensive rats. This study examined (i) whether the autonomic nervous system is involved in the mediation of 1,8-cineole-induced changes in mean aortic pressure (MAP) and heart rate (HR) and (ii) whether the hypotensive effects of 1,8-cineole could result from its vasodilatory effects directly upon vascular smooth muscle. In both pentobarbital-anesthetized and conscious, freely moving rats, bolus injections of 1,8-cineole (0.3-10 mg/kg, i.v.) elicited similar and dose dependent decreases in MAP. Concomitantly, 1,8-cineole significantly decreased HR only at the highest dose (10 mg/kg). Pretreatment of anesthetized rats with bilateral vagotomy significantly reduced the bradycardic responses to 1,8-cineole (10 mg/kg) without affecting hypotension. In conscious rats, i.v. pretreatment with methylatropine (1 mg/kg), atenolol (1.5 mg/kg), or hexamethonium (30 mg/kg) had no significant effects on the 1,8-cineole-induced hypotension, while bradycardic responses to 1,8-cineole (10 mg/kg) were significantly reduced by methylatropine. In rat isolated thoracic aorta preparations, 1,8-cineole (0.006 2.6 mM) induced a concentration-dependent reduction of the contraction induced by potassium (60 mM). This is the first physiological evidence that i.v. treatment with 1,8-cineole in either anesthetized or conscious rats elicits hypotension; this effect seems related to an active vascular relaxation rather than withdrawal of sympathetic tone. PMID- 12564638 TI - Cyclic nucleotide phosphodiesterase isozymes expressed in mouse skeletal muscle. AB - To understand changes in cyclic nucleotide metabolism in muscle disease states, the expression of phosphodiesterase (PDE) isozymes in normal mouse leg muscle was examined. Four subcellular fractions were generated by differential centrifugation at 10,000 x g and 100,000 x g. cAMP PDE activity was found predominately in the soluble fractions, while cGMP PDE activity was more evenly distributed amongst soluble and particulate fractions. Pharmacological inhibitors demonstrate that PDE4 represents the major cAMP hydrolyzing activity and PDE2 represents the major cGMP hydrolyzing activity in mouse leg muscle. PDE1 is expressed at low levels, while PDE3 and PDE5 are intermediate. Between 20 and 40% of total PDE activity remained in the presence of inhibitors for PDE1-PDE5, indicating that other PDE families contribute to the total PDE pool. Reverse transcription PCR with family-specific primers showed expression of mRNA for PDE7 PDE9, supporting this conclusion. Total PDE activity was found to be elevated in tissue extracts from a mouse model of Duchenne's muscular dystrophy. PMID- 12564639 TI - Peripheral chemoreceptor contributions to sympathetic and cardiovascular responses during hypercapnia. AB - We tested the hypothesis that integrated sympathetic and cardiovascular reflexes are modulated by systemic CO2 differently in hypoxia than in hyperoxia (n = 7). Subjects performed a CO2 rebreathe protocol that equilibrates CO2 partial pressures between arterial and venous blood and that elevates end tidal CO2 (PET(CO2)) from approximately 40 to approximately 58 mmHg. This test was repeated under conditions where end tidal oxygen levels were clamped at 50 (hypoxia) or 200 (hyperoxia) mmHg. Heart rate (HR; EKG), stroke volume (SV; Doppler ultrasound), blood pressure (MAP; finger plethysmograph), and muscle sympathetic nerve activity (MSNA) were measured continuously during the two protocols. MAP at 40 mmHg PET(CO2) (i.e., the first minute of the rebreathe) was greater during hypoxia versus hyperoxia (P < 0.05). However, the increase in MAP during the rebreathe (P < 0.05) was similar in hypoxia (16 +/- 3 mmHg) and hyperoxia (17 +/- 2 mmHg PET(CO2)). The increase in cardiac output (Q) at 55 mmHg PET(CO2) was greater in hypoxia (2.61 +/- 0.7 L/min) versus hyperoxia (1.09 +/- 0.44 L/min) (P < 0.05). In both conditions the increase in Q was due to elevations in both HR and SV (P < 0.05). Systemic vascular conductance (SVC) increased to similar absolute levels in both conditions but rose earlier during hypoxia (> 50 mmHg PET(CO2)) than hyperoxia (> 55 mmHg). MSNA increased earlier during hypoxic hypercapnia (> 45 mmHg) compared with hyperoxic hypercapnia (> 55 mmHg). Thus, in these conscious humans, the dose-response effect of PET(CO2) on the integrated cardiovascular responses was shifted to the left during hypoxic hypercapnia. The combined data indicate that peripheral chemoreceptors exert important influence over cardiovascular reflex responses to hypercapnia. PMID- 12564640 TI - Specific electromechanical responses of cardiomyocytes to individual and combined components of ischemia. AB - The main factors of myocardial ischemia are hypoxia, substrate deprivation, acidosis, and high extracellular potassium concentration ([K+]e), but the influence of each of these factors has not yet been evaluated in a cardiomyocyte (CM) culture system. Electromechanical responses to the individual and combined components of ischemia were studied in CM cultured from newborn rat ventricles. Action potentials (APs) were recorded using glass microelectrodes and contractions were monitored photometrically. Glucose-free hypoxia initially reduced AP duration, amplitude, and rate and altered excitation-contraction coupling, but AP upstroke velocity (Vmax) remained unaffected. Early afterdepolarizations appeared, leading to bursts of high-rate triggered impulses before the complete arrest of electromechanical activity after 120 min. Acidosis reduced Vmax whereas AP amplitude and rate were moderately decreased. Combining acidosis and substrate-free hypoxia also decreased Vmax but attenuated the effects of substrate-free hypoxia on APs and delayed the cessation of the electrical activity (180 min). Raising [K+]e reduced the maximal diastolic potential and Vmax. Total ischemia (substrate deletion, hypoxia, acidosis, and high [K+]e) decreased AP amplitude and Vmax without changing AP duration. Moreover, delayed afterdepolarizations appeared, initiating triggered activity. Ultimately, 120 min of total ischemia blocked APs and contractions. To conclude, glucose-free hypoxia caused severe functional defects, acidosis delayed the changes induced by substrate-free hypoxia, and total ischemia induced specific dysfunctions differing from those caused by the former conditions. Heart-cell cultures thus represent a valuable tool to scrutinize the individual and combined components of ischemia on CMs. PMID- 12564641 TI - Effects of clozapine on the efflux of serotonin and dopamine in the rat brain: the role of 5-HT1A receptors. AB - In vivo microdialysis in conscious rats was used to examine the effect of clozapine on serotonin (5-hydroxytryptamine, 5-HT) efflux in the prefrontal cortex and dorsal raphe nucleus and dopamine efflux in the prefrontal cortex. Both systemic and local administration of clozapine (systemic, 10 or 20 mg/kg, i.p.; local, 100 microM) increased 5-HT efflux in the dorsal raphe. However, in the prefrontal cortex, dialysate 5-HT increased when clozapine (100 microM) was administered through the probe, while no effect was observed when it was administered systemically. By pretreatment with the selective 5-HT1A receptor antagonist p-MPPI (3 mg/kg, i.p.), systemic treatment of clozapine (10 mg/kg, i.p.) significantly increased 5-HT efflux in the prefrontal cortex. This result suggests that the ability of clozapine to enhance the extracellular concentrations of 5-HT in the dorsal raphe attenuates this drug's effect in the frontal cortex, probably through the stimulation of 5-HT1A somatodendritic autoreceptors in the dorsal raphe. We also found that pretreatment with p-MPPI (3 mg/kg, i.p.) attenuated by 45% the rise in cortical dopamine levels induced by clozapine (10 mg/kg, i.p.). These findings imply that the reduction in serotonergic input from the dorsal raphe nucleus induced by clozapine could lead to an increase in dopamine release in the prefrontal cortex. PMID- 12564642 TI - Models of depressed hepatic mrp2 activity reveal bromosulphophthalein-sensitive passive K+ flux. AB - Bile acid independent flow composes up to 40% of hepatic bile secretory capacity. Apical (canalicular) efflux of non-bile-acid organic anions provides the major osmotic driving force for bile acid independent flow. Organic anion accumulation in the hepatocyte is accompanied by increases in both K+ conductance in isolated hepatocytes and passive K+ flux in the perfused rat liver, which are indicative of K+ channel activation. We used two models of disrupted canalicular anion transport to test whether organic anion stimulated K+ efflux occurs independently of anion excretion. In both wild type (wt) and mrp2 mutant (transport minus, tr-) rat liver, bromosulfophthalein (BSP; 0.5 mM) caused a reversible increase in K+ flux that (i) was outwardly directed with low external K+ and (ii) depended upon the electrochemical potential for K+. K+ efflux from wt livers of both sexes was about 1.5 times larger than that from tr- livers. Further, K+ release from female rat livers was about three times higher than that from male livers, independent of phenotype. Two transcripts of the rat hepatocyte K+ channel (Kir4.2) were expressed in hepatocytes of all rats. The results demonstrate that BSP stimulates basolateral (sinusoidal) K+ channels independently of its canalicular excretion, revealing an early event in BAIF and suggesting that Kir4.2 may mediate BSP sensitive K+ flux. PMID- 12564643 TI - Carbachol interactions with nonsteroidal anti-inflammatory drugs. AB - The inhibition of cyclooxygenase enzymes by nonsteroidal anti-inflammatory drugs (NSAIDs) does not completely explain the antinociceptive efficacy of these agents. It is known that cholinergic agonists are antinociceptive, and this study evaluates the interactions between carbachol and some NSAIDs. Antinociceptive activity was evaluated in mice by the acetic acid writhing test. Dose-response curves were constructed for NSAIDs and carbachol, administered either intraperitoneally (i.p.) or intrathecally (i.t.). The interactions of carbachol with NSAIDs were evaluated by isobolographic analysis after the simultaneous administration of fixed proportions of carbachol with each NSAID. All of the drugs were more potent after spinal than after systemic administration. The combinations of NSAIDs and carbachol administered i.p. were supra-additive; however, the i.t. combinations were only additive. Isobolographic analysis of the coadministration of NSAIDs and carbachol and the fact that atropine antagonized the synergistic effect suggest that carbachol may strongly modulate the antinociceptive activity of NSAIDs; thus, central cholinergic modulation would be an additional mechanism for the antinociceptive action of NSAIDs, unrelated to prostaglandin biosynthesis inhibition. PMID- 12564644 TI - Pressor responsiveness to angiotensin in soy-fed spontaneously hypertensive rats. AB - Dietary soy may attenuate the development of arterial hypertension. In addition, some soy-containing foods exhibit angiotensin-converting enzyme (ACE) inhibitory properties. Accordingly, we tested the hypothesis that ACE inhibition contributes to the antihypertensive effect of dietary soy. Mean arterial blood pressure (MAP) was recorded from conscious spontaneously hypertensive rats (SHR) at least 24 h after the implantation of catheters. Cumulative dose-response curves to intravenous angiotensin I (AI) (5-100 ng x kg(-1) x min(-1)) and angiotensin II (AII) (1-20 ng x kg(-1) x min(-1)) were constructed for male, sham-operated female, and ovariectomized female (OVX) SHR that were maintained on either casein or soy diets. The soy diet was associated with a significant reduction in baseline MAP in the OVX SHR (approximately 20 mmHg, 1 mmHg = 133.322 Pa). AI and AII infusions caused graded increases in MAP in all groups. However, there was no significant attenuation of the pressor responses to AI in the soy-fed SHR. Conversely, we observed a significant rightward displacement of the AII dose response curves in the soy-fed sham-operated and OVX SHR. We conclude that ACE inhibition does not account for the antihypertensive effect of dietary soy in mature SHR. PMID- 12564645 TI - Antioxidative and free radical scavenging effects of ecdysteroids from Serratula strangulata. AB - The antioxidative and free radical scavenging effects of four ecdysteroids, 20 hydroxyecdysone (E1), 25-deoxy-11,20-dihydroxyecdysone (E2), 24-(2-hydroxyethyl) 20-hydroxyecdysone (E3), and 20-hydroxyecdysone-20,22-monoacetonide (E4), isolated from the Chinese herb Serratula strangulata have been investigated in vitro. These ecdysteroids could protect human erythrocytes against oxidative hemolysis induced by a water-soluble azo initiator 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH). They could also inhibit the peroxidation of rat liver microsomes induced by hydroxyl radicals, as monitored by the formation of thiobarbituric acid reactive substances (TBARS), and prevent radical-induced decrease of membrane fluidity as determined by fluorescence polarization. They reacted with galvinoxyl radicals in homogeneous solution, and the pseudo-first order rate constants were determined spectrophotometrically by following the disappearance of galvinoxyl radicals. Compounds E1 and (or) E3 were the most active in both antioxidative and radical-scavenging reactions. PMID- 12564646 TI - Thyroid-hormone-induced phosphate absorption in Aplysia californica gut is mediated through protein synthesis. AB - The mucosal membranes of Aplysia californica forgut epithelia contain a sodium phosphate symporter that is stimulated by triiodothyronine. Actinomycin D, puromycin, or cycloheximide inhibited the triiodothyronine-stimulated phosphate absorption. It appears that thyroid hormone manifests its effects on phosphate absorption in the A. californica gut through protein synthesis. PMID- 12564647 TI - Vitamin C lowers blood pressure and alters vascular responsiveness in salt induced hypertension. AB - The present study was undertaken to investigate the effect of vitamin C treatment on blood pressure and vascular reactivity in salt-induced hypertension. Male Sprague-Dawley rats were fed a normal rat diet, a high-sodium (8% NaCl) diet, a normal rat diet plus vitamin C treament (100 mg x kg(-1) x day(-1)), or a high sodium diet plus vitamin C treatment for 6 weeks. Salt loading significantly increased blood pressure, which was attenuated by vitamin C treatment. Aortic rings from the different groups were suspended for isometric-tension recording. The contractile response to noradrenaline was significantly increased in the salt loaded rats. Vitamin C reduced the sensitivity of aortic rings to noradrenaline in rats on normal and high-sodium diets. In noradrenaline-precontracted rings, the relaxation response to acetylcholine, which was attenuated in the salt-loaded rats, was restored by vitamin C treatment. Pretreatment with N(G)-nitro-L arginine methyl ester (L-NAME) abolished the enhanced response to acetylcholine caused by vitamin C. The results suggest that the antihypertensive effect of vitamin C is associated with a reduction in vascular sensitivity to noradrenaline and enhancement of endothelium-dependent relaxation due to increased nitric oxide bioavailability. PMID- 12564648 TI - Inhibitory effect of a novel bradykinin B1 receptor antagonist, R-954, on enhanced vascular permeability in type 1 diabetic mice. AB - The morbidity and mortality associated with type 1 diabetes are essentially related to the micro- and macrovascular complications that develop over time and lead to several diabetic complications, including hypertension, atherosclerosis, and retinopathy, as well as coronary and renal failure. Normally absent in physiological conditions, the bradykinin B1 receptor (BKB1-R) was recently found to be overexpressed in pathological conditions, including type 1 diabetes. In the present study, we evaluated the effect of the new BKB1-R antagonist, R-954 (Ac Orn-[Oic2, alpha-MePhe5, D-betaNal7, Ile8]desArg9-bradykinin, on the increase in vascular permeability in streptozotocin (STZ)-diabetic mice. The capillary permeability to albumin was measured by quantifying the extravasation of albumin bound Evans blue dye in selected target tissues (liver, pancreas, duodenum, ileum, spleen, heart, kidney, stomach, skin, muscle, and thyroid gland). Acute single administration of R-954 (300 microg/kg, i.v.) to type 1 diabetic mice 4 weeks after STZ significantly inhibited the enhanced vascular permeability in most tissues. These data provide further experimental evidence for the implication of BKB1-R in the enhanced vascular permeability associated with type 1 diabetes. PMID- 12564649 TI - Staying in hospital social work. AB - Hospitals have traditionally been large employers of social workers. As workplaces they are often identified as being challenging and ambiguous environments for social work practice. In this study the experience of hospital work is explored. Social workers' 'tolerance' of the hospital environment is identified as a way of explaining 'staying.' Staying has both positive and negative features which arise from the interaction between workers' tolerance of the environment and their qualities of 'self-actualisation.' The identification of this relationship informs and challenges existing theories of retention and turnover of workers. PMID- 12564650 TI - Assessing the practical and psychosocial needs of rural women with early breast cancer in Australia. AB - The purpose of the current study was to assess the practical and psychosocial needs of rural women with early breast cancer in Australia and recommend strategies to ensure equity in availability and access to cancer treatment for all women. A random sample of 204 rural women diagnosed with early breast cancer was recruited to participate in a telephone survey via state and territory cancer registries. Although the majority of women were satisfied with their provision of information overall, less than a third of participants were provided with specific information on assistance for rural women. Findings also revealed that only 47% of the women who had to travel for treatment received financial assistance, and 13% of these women had difficulty organizing or claiming financial assistance. Furthermore, only 10% of women found social workers to be a source of support. PMID- 12564651 TI - Pregnancy loss: what it means to women who miscarry and their partners. AB - This paper reports results from six women who had miscarried and their partners in Melbourne. A qualitative approach, namely in-depth semi-structured interviews, was utilised to obtain information. Several themes emerged from the data. The findings of this study show that women and men experienced a variety of feelings in reaction before, during and after the miscarriage. These feelings varied in type, intensity, and duration in which the men generally experienced less intense feelings for a shorter period of time as compared to the women. We argue that support and counselling be provided not only for women who miscarry, but also for their male partner, as the men may have fewer means to express their grief due to the "social expectation" of male roles. PMID- 12564652 TI - Emotional dissonance in medical social work practice. AB - This qualitative study examined several ecological aspects of medical social work practice that affect social workers' emotional well-being. Forty-seven medical social workers from seven hospitals participated in small group interviews in which practice experiences were explored. A grounded theory approach was used to analyze the data, with emotional dissonance emerging as a central theme. Community, family, and individual factors contributing to emotional dissonance are presented, and the effect of social support is examined. PMID- 12564653 TI - Access to services and maintenance of safer sex practices among people living with HIV/AIDS. AB - Access to services and their relationship to the maintenance of long-term safer sex practices are addressed in this study of 360 HIV+ adults recruited from outpatient medical facilities. Protease inhibitors, antiviral therapies, and entitlements were reported as the most needed services, while entitlements and money to pay for housing were reported as the largest unmet needs. Differences across ethnic and gender groups were observed. One-third of all respondents reported at least one occasion of unprotected anal or vaginal intercourse in the previous six months. The practice of unsafe sex was found to be significantly related to both the number of needed services and the number of unmet needs, even after controlling for demographic variables. In addition, a higher proportion of those who engaged in unsafe sex reported a higher need for psychological counseling and social support. These findings underscore the important linkage between access to services with avoidance of high-risk sexual behavior in HIV+ persons. Implications for the delivery of culturally appropriate, gender-specific and community-based interventions are discussed. PMID- 12564654 TI - Formoterol delivered via a dry powder inhaler (Aerolizer): results from long-term clinical trials in children. AB - Over 500 children with asthma, aged 5-12 years, have been treated with formoterol fumarate (Foradil) delivered via the Aerolizer dry powder inhaler in clinical trials, with treatment periods of up to 15 months. In pivotal double-blind trials, two dose levels, 12 and 24 microg taken twice daily, provided significant benefit in terms of lung function measurements and symptom control (a lower dose of 6 microg twice daily appeared insufficient with this formulation). The higher, 24 microg dose appeared to provide an additional margin of benefit in a subgroup of children with more unstable/severe disease when the results from long-term follow-up (12-15 months) were analysed. Formoterol was shown to have a good safety profile when taken as regular maintenance treatment and when used as rescue medication by patients already receiving formoterol as regular maintenance treatment. In this flexible regimen, with formoterol used for rescue and maintenance, the overall daily intake of formoterol was low, with 96.1% of all treatment days (n = 2452) covered by a total daily dose (regular + rescue) of 48 microg (four doses) or less. There was no increase in the average daily intake of rescue formoterol over time. The clinical efficacy associated with this regimen was maintained over time and, in the case of morning peak expiratory flow rate, steadily improved over time. The Foradil Aerolizer inhalation system is simple to use and has a low resistance to inspiratory airflow that maximises the patient's control over dosing, while minimising the risk of under- and overdosing. These features may be especially valuable in a young patient population. PMID- 12564655 TI - An international study of the effects of rosiglitazone plus sulphonylurea in patients with type 2 diabetes. AB - This was an open-label, randomised 26-week study to determine the effects of adding 4 mg rosiglitazone (Avandia) daily to existing sulphonylurea (SU) therapy in patients with type 2 diabetes from India, Brazil, The Philippines, Thailand, Argentina and Tunisia. Of the 348 patients, 175 received 2 mg rosiglitazone twice daily plus SU (RSG+SU) and 173 received SU alone (at their normal dose). The RSG+SU group showed a significant reduction in HbA1c (mean HbA1c 9.05% at baseline, 7.92% at 26 weeks, mean change -1.13 (95% Cl -1.37, -0.89)). Mean HbA1c essentially remained unchanged in the control group (8.9 to 9.0%). The RSG+SU group showed a significant decrease in fasting plasma glucose concentration (FPG) (mean FPG 198.7 mg/dl at baseline, 160.3 mg/dl at 26 weeks, mean change -38.4 (95% Cl -47.1, -29.7)) while the controls showed a non-significant increase from 194 to 200 mg/dl. Significantly more patients in the RSG+SU group achieved FPG < 140 mg/dl, > or = 0.7% decrease in HbA1c, and > or = 30 mg/dl decrease in FPG between baseline and week 26 than the controls (p = 0.0001 in each case). Adverse events were similar in both groups; more patients in the RSG+SU group reported hypoglycaemia, but most cases were mild. This study shows that adding rosiglitazone to existing SU treatment improves glycaemic control and is well tolerated in patients with type 2 diabetes from a wide range of non-Western countries. PMID- 12564656 TI - The prevalence and treatment of headache sufficient to impact on the quality of life of undergraduate students entering university. AB - AIM OF STUDY: Headache has a significant impact on public health in terms of quality of life and economic consequences, but in primary care, needs often remain unmet in terms of recognition, diagnosis and treatment. Our aim was to measure the prevalence of headache sufficient to affect the quality of life of undergraduate students who were entering the University of Exeter. METHOD: 1124 consecutive undergraduate entrants aged 21 and under who registered at the Student Health Centre at the beginning of the academic year were invited to complete a questionnaire during their registration procedure. We used the Headache Impact Score to measure effect of headache on quality of life. RESULTS: A completed questionnaire was received from 1057 (94%) students registering at the Health Centre. 212 (21%) of these students had headaches that affected the quality of their life. Of these, 95 (45%) had seen a doctor previously about their headaches and 28 (13%) had headaches on more than 15 days a month. Less than 5% were taking prescription medication. The headache impact score was 56 indicating a substantial impact on quality of life of students. CONCLUSION: Headache has a considerable impact on the quality of life of students entering University which we speculate may have a deleterious effect on educational attainment. This study has confirmed the findings of other population groups that morbidity from headache is often unrecognised and under treated. PMID- 12564657 TI - Calcitonin gene-related peptide dilates the pregnant rat uterine vascular bed via guanylate cyclase, ATP- and Ca-sensitive potassium channels and gap junctions. AB - We studied the mechanism of calcitonin gene-related peptide (CGRP)-induced vasorelaxation in isolated uterine vascular beds of pregnant rats. The vascular beds were perfused in situ with Krebs buffer containing dextran and indomethacin, an inhibitor of cyclooxygenase. Baseline perfusion pressure was maintained with norepinephrine. When applied as a bolus, CGRP caused a decreased perfusion pressure in uterine vascular beds that was dose-dependent and equal in both mid pregnant and late-pregnant rats. The non-selective inhibitor of nitric oxide synthase (NOS), Nomega-nitro-L-arginine methyl ester (L-NAME), did not significantly affect CGRP-induced vasodilatation in vascular beds of either group. CGRP-induced vasodilatation was not influenced by preincubation with the inhibitors of adenylate cyclase (SQ 22536 or MDL 12330A), but was significantly attenuated by the selective inhibitor of soluble guanylate cyclase (ODQ). The vasorelaxant effect of CGRP was not significantly influenced by the inhibitor of voltage-gated potassium (KV) channels (4-aminopyridin), but was significantly attenuated by an inhibitor of calcium-regulated potassium (KCa) channels (tetraethylammonium) and by an inhibitor of adenosine triphosphate-sensitive potassium (KATP) channels (glibenclamide). The gap junction uncoupling agent (carbenoxolone) also significantly attenuated the CGRP-induced decrease in perfusion pressure. We conclude that vasorelaxation induced by CGRP in the pregnant rat uterine vascular bed is not dependent on endothelial nitric oxide. In the uterine circulation of late-pregnant rats, the CGRP effect involves activation of soluble guanylate cyclase, but not adenylate cyclase, and does involve KCa and KATP channels and gap junctions. PMID- 12564658 TI - Management by nurses of primary headache: a pilot study. AB - Primary headache is a very common condition and one that nurses encounter in many different care settings. Yet there is a lack of evidence as to whether advice given to sufferers is effective and what improvements may be expected in the condition. The aim of this study was to evaluate the advice given by a nurse to primary headache sufferers. The design was quasi-experimental. An experimental group (n = 19) and a control group (n = 25) of primary headache sufferers had their headache parameters of frequency, severity duration and disability (Migraine Disability Assessment) over the previous six months assessed. The experimental group then received advice in the form of health education from a nurse. Both groups kept a headache diary for six months. After six months both groups had their headache parameters assessed again. Compared to the control group, there was a significant reduction in the severity of headaches experienced by the experimental group (p = 0.006). Although there were reductions in frequency and duration of headaches experienced by the experimental group compared to the control group, these were not significant (p = 0.664 and p = 0.235, respectively). The Migraine Disability Assessment showed a trend towards reduced scores in the experimental group compared to controls which were not significant (p = 0.535). This pilot study suggests that health education can be effective in reducing the severity of headaches. However, a larger study over a longer period is needed to evaluate improvements in headache parameters. PMID- 12564659 TI - Cilostazol treatment of claudication in diabetic patients. AB - OBJECTIVE: To compare the efficacy and safety of cilostazol in diabetic and non diabetic patients from eight (six placebo- and two active-controlled) randomised, double-blind phase III trials. DESIGN: We only included patients from the trial data set receiving cilostazol 100 mg twice daily (216 diabetic/599 non-diabetic) or placebo (220/616). Efficacy was measured by absolute claudication distance (ACD), using standard treadmill exercise protocols. RESULTS: Among diabetic and non-diabetic patients, cilostazol was superior to placebo (estimated treatment effect 1.15, 95% confidence interval, 1.05-1.25, p = 0.001; and 1.24, 1.18-1.31, p < 0.0001, respectively). There was no statistical difference in response between diabetic and non-diabetic subjects. In the efficacy analysis, cilostazol treated diabetic subjects with the lowest baseline ACD (but not those with greater baseline ACD) walked approximately 34% farther than at baseline, whereas their non-diabetic counterparts walked 23% farther. There was no significant difference in the adverse event profile of the diabetic and non-diabetic patients on cilostazol. No excess haemorrhagic events occurred in cilostazol-treated diabetic patients. Trial duration varied from 12 to 24 weeks. CONCLUSIONS: Diabetic and non-diabetic patients with intermittent claudication respond favourably to cilostazol, with no significant difference in their overall response. Diabetic individuals with the most severe claudication respond better than those less affected, but the response of non-diabetic patients increases as baseline ACD increases. Adverse event incidence was comparable in the two populations, although diabetic patients might be expected to experience greater morbidity. Cilostazol is a safe and effective treatment for claudication in diabetic and non-diabetic populations. PMID- 12564660 TI - Longitudinal follow-up of TTS-fentanyl use in patients with cancer-related pain: results of a compassionate-use study with special focus on elderly patients. AB - GOALS OF THE WORK: This open compassionate-use prospective registration study evaluated the tolerability, ease of use and applied doses of transdermal (TTS) fentanyl in adult patients with cancer-related pain requiring strong opioid analgesia. Elderly patients were particularly focussed on. PATIENTS AND METHODS: Previous pain medication was converted to an appropriate dose of TTS-fentanyl. Immediate-release morphine rescue medication was allowed as needed for breakthrough pain. Dose adjustments of TTS-fentanyl, rescue morphine requirements, the ease of use and side-effects were assessed monthly, with special emphasis paid to the severity of constipation and the use of laxatives. MAIN RESULTS: A total of 663 patients with cancer-related pain, including 8% opioid-naive patients, were enrolled; 661 patients used at least 1 patch of TTS fentanyl. Of these, 455 subjects were assessed at baseline and at 1 post-baseline visit at least. Individual treatment ranged from a few days to 2 1/2 years; TTS fentanyl doses ranged from 25 to 950 microg/h. The major reason for study termination was non-drug-related death (61%). Approximately 40% of patients reported constipation. The frequency of constipation depended on the rescue morphine dose used, but no dose-relationship was found for TTS-fentanyl. Patient acceptance of the patches was high; around 85% of patients rated convenience as good to excellent The ease of use and tolerability of TTS-fentanyl in the elderly patients were comparable to that in the total population, except for a slight increase of non-serious adverse events. CONCLUSIONS: TTS-fentanyl can be applied as long-term therapy to patients with cancer-related pain, including the elderly. PMID- 12564661 TI - Attaining United Kingdom-European Atherosclerosis Society low-density lipoprotein cholesterol guideline target values in the Greek Atorvastatin and Coronary-heart disease Evaluation (GREACE) Study. AB - The GREek Atorvastatin and Coronary-heart-disease Evaluation (GREACE) Study compared two standards (structured vs. usual care) of lipid lowering treatment in 1600 patients with coronary heart disease (CHD). Structured care aimed at achieving (with atorvastatin 10-80 mg) the low-density lipoprotein cholesterol (LDL-C) (2.6 mmol/l; 100 mg/dl) goal described in the NCEP ATP II and III guidelines for patients with CHD. Structured care was associated with a significant reduction in overall mortality and coronary events compared to usual care. In the present brief report we interpret the results of GREACE using the United Kingdom (UK) and European Atherosclerosis Society (EAS) treatment goal for LDL-C in secondary CHD prevention (3.0 mmol/l; 115 mg/dl. The mean dose of atorvastatin decreased from 24 mg to 22 mg/day. More patients achieved the UK and EAS LDL-C target (95.6 vs. 95%) in the structured care arm of the trial; 90% of the patients achieved this target with 10 or 20 mg atorvastatin. These findings may have cost implications, especially if the LDL-C target for high-risk patients will fall below those described above. PMID- 12564662 TI - The biochemical selectivity of novel COX-2 inhibitors in whole blood assays of COX-isozyme activity. AB - We have evaluated the biochemical selectivity of novel cyclo-oxygenase (COX)-2 inhibitors, etoricoxib, valdecoxib, DFU and DFP, vs rofecoxib and celecoxib, using the human whole blood assays of COX-isozyme activity, in vitro. Compounds were incubated with human whole blood samples, allowed to clot for 1 h at 37 degrees C, or stimulated with lipopolysaccharide (10 microg/ml) for 24 h at 37 degrees C. Serum thromboxane (TX) B2 and plasma prostaglandin (PG) E2 levels were measured by specific radioimmunoassays as indices of platelet COX-1 and monocyte COX-2 activity, respectively. Valdecoxib, etoricoxib, DFU and DFP inhibited platelet COX-1 and monocyte COX-2 with the following COX-1/COX-2 IC50 ratios: 61.5, 344, 660 and 1918, respectively. The reference compounds, celecoxib and rofecoxib had corresponding values of 29.6 and 272. In conclusion, a second wave of COX-2 inhibitors with higher biochemical selectivity than the existing coxibs has been developed. Whether their administration will be associated with improved clinical efficacy and/or safety vis-a-vis celecoxib and rofecoxib remains to be established. PMID- 12564663 TI - Impact of montelukast on symptoms in mild-to-moderate persistent asthma and exercise-induced asthma: results of the ASTHMA survey. Adding Singulair Treatment to Handle symptoms in Mild to moderate Asthmatics. AB - A nation-wide survey was undertaken in Belgium among general practitioners (GPs) to evaluate the impact of the leukotriene receptor antagonist (LTRA) montelukast on the control of asthma symptoms, after at least 4 weeks of treatment. Patients from 6 years of age were eligible if they were suffering from mild-to-moderate persistent asthma which was still symptomatic despite inhaled corticosteroids (ICS) treatment, or from exercise-induced asthma. Patient general satisfaction was evaluated by recording the willingness to continue the treatment. A total of 1360 GPs took part in the study and more than 11000 patients were included in the survey. Of the included patients, 85% were receiving inhaled corticosteroids, 60% of whom were also on long-acting beta2-agonists (LABA). However, despite the use of daily controller medication, 92% of the patients still reported limitation of activities, 49% difficulties with sleep and 45% early morning awakening due to asthma. Moreover, 78% of the patients used rescue medication more than twice a week. At the end of the survey, 90% of the patients expressed their willingness to continue montelukast therapy. Of the patients having symptoms at the start of the study, 87% reported amelioration in sleep while on montelukast therapy, 80% less frequent early morning awakening, 85% better ability to perform daily activities and 77% decreased need for rescue medication. PMID- 12564664 TI - Comparative efficacy of two anti-bacterial/anti-inflammatory formulations (Auricularum otic powder and Dex-Otic drops) in the medical treatment of otitis externa. AB - BACKGROUND: Otitis externa (OE) is a common disorder which can cause severe pain and discomfort with discharge and hearing loss. The majority of topical preparations for OE treatment contain a combination of steroids and antibiotics and are administered as ear drops. Otic powder preparations are less common. We found no studies which compared the efficacy of the two dosage forms. OBJECTIVE: The purpose of the study was to compare the healing process of the external ear canal when treated locally either with otic powder (Auricularum), a preparation containing antibiotic, steroidal and antimycotic components, or with otic drops (Dex-Otic), a commercial ear drop preparation containing antibiotic and steroidal components. METHODS: The study was designed as an open, randomised, comparative clinical trial. Ambulatory patients suffering from OE were examined by an ear specialist and randomly divided into an otic powder (Auricularum) treatment group and an ear drop (Dex-Otic) treatment group. Each treatment group was treated in accordance with the daily doses recommended by the manufacturers. Patients were treated for a period of 14 days and examined on days 0, 3, 7, 10 and 14. Clinical signs and symptoms recorded were pain, erythema, swelling and discharge. RESULTS: A total of 67 patients entered the study. The probability of healing within 7 days was 74% in the otic powder group as opposed to 40% in the ear drops group (log rank test, p = 0.0013). The probability of total pain relief after 3 days of treatment was 60% in the otic powder group compared to 53% in the ear drops group (log rank test, p = 0.0001). CONCLUSION: We conclude that treatment with an otic powder (Auricularum) may promote earlier healing and pain relief than does treatment with otic drops (Dex-Otic). PMID- 12564665 TI - Intergenerational transmission of health: the role of intrapersonal, interpersonal, and communicative factors. AB - A model of household dietary behaviors based on adults' and children's intrapersonal, interpersonal, and communicative factors is introduced and tested. To the extent that children's health behaviors are influenced by adults' health behaviors, it is hypothesized that children's behavioral determinants, including self-efficacy, knowledge, and use of health information, will be affected by household adults' behavioral determinants. Household communication is expected to affect the behaviors and the behavioral determinants of adults and children. Data come from the Stanford Five-City Project. Structural equation models revealed that adults' dietary behavior was influenced by their self-efficacy, knowledge, and discussion between adults and children. Children's dietary behavior was influenced by their self-efficacy, knowledge, and use of health information. Adults' intrapersonal determinants of dietary behavior predicted corresponding children's measures. Implications for health education efforts directed at children include encouraging household discussion about health and focusing on adults as agents of change as an integral campaign strategy. PMID- 12564666 TI - Gender differences in young adults' beliefs about sunscreen use. AB - This study employs focus group methodology to explore gender differences in sunscreen use. Guided by the theory of reasoned action, males and females were found to differ on each of the following constructs: behavior, behavioral beliefs, and normative beliefs. Males and females differed in their sunscreen use, with females adopting a more preventive style of sunscreen use and males a more reactive style. Males and females differed in their salient beliefs that motivated their sunscreen use, many of which were related to traditional American gender roles. In addition, although males and females were aware of both positive and negative sources of normative beliefs regarding sunscreen use, females received more encouragement from their mothers and peers than males. Findings are discussed in terms of their implications for the design of future interventions. PMID- 12564667 TI - Intervention programs for arthritis and other rheumatic diseases. AB - Disability reduction or prevention programs for people with arthritis and other rheumatic conditions reduce long-term pain and disability but reach only a fraction of their target audience. Few public health professionals are aware of these programs or their benefits. The objective of this study is to review and describe packaged (ready-to-use) arthritis self-management education and exerciselphysical activity programs that have had at least preliminary evaluation. Nine intervention programs (five self-management education programs, and four exercise/physical activity programs met study criteria). Several of the packaged arthritis interventions reviewed help people with arthritis and other rheumatic conditions maximize their abilities and reduce pain, functional limitations, and other arthritis-related problems. Other packaged interventions show promise in reducing pain, disability, and depression and in increasing self care behaviors, but they need to be evaluated more extensively. PMID- 12564668 TI - A content analysis of Web sites promoting smoking culture and lifestyle. AB - The present study examined smoking culture and lifestyle Web sites listed on Yahoo!, a popular Internet search catalog, to determine whether the sites were easily accessible to youth, featured age or health warnings, and mentioned specific tobacco brands. A content analysis of photographs on these sites assessed the demographics of individuals depicted and the amount of smoking and nudity in the photographs. The sample included 30 Web sites, all of which were accessible to youth and did not require age verification services to enter them. Cigarette brand names were mentioned in writing on 35% of the sites, and brand images were present on 24% of the sites. Nearly all of the photographs (95%) depicted smoking, 92% featured women, and 7% contained partial or full nudity. These results underscore the need for greater research and monitoring of smoking related Internet content by health educators and tobacco control advocates. PMID- 12564669 TI - "If they don't ask about condoms, I just tell them": a descriptive case study of Latino lay health advisers' helping activities. AB - Health care practitioners have increasingly turned to lay health advisers (LHAs) to deliver health promotion and disease prevention supportive activities to underserved populations. Yet, published evaluations of the LHA programs or the manner in which LHAs deliver information are rare. An important aspect of program evaluation includes understanding the role of the LHA in the community as described by the LHAs in the field. The purpose of this article is to report the results of a component of a larger evaluation of an LHA program for preventing HIV/AIDS among recently immigrated Mexicans in North Carolina. Specifically, the authors use a descriptive case study design to report on the perceptions of the LHAs related to program objectives. The findings suggest that the LHAs are meeting the program objectives and are confident in their role as LHAs. PMID- 12564670 TI - Promoting repeat mammography use: insights from a systematic needs assessment. AB - This article describes the process and outcome of a needs assessment conducted to guide the development of interventions to increase repeat mammography use among participants in a federally funded cancer screening program. Health behavior theory and data from a phone survey are used to uncover key barriers to repeat mammography use and to identify fruitful intervention approaches for modifying them. Estimates of (a) compliance with mammography guidelines, (b) readiness to adopt regular mammography use, (c) the most common reasons for not being rescreened, and (d) population attributable risks associated with various predictors of repeat mammography use are presented and, with guidance from the transtheoretical model of behavior change, used to make inferences about the type of intervention strategies most appropriate for promoting repeat mammography use in this population. PMID- 12564671 TI - Solid-phase microextraction for the analysis of short-chain chlorinated paraffins in water samples. AB - A novel solid-phase microextraction (SPME) method coupled to gas chromatography with electron capture detection (GC-ECD) was developed as an alternative to liquid-liquid and solid-phase extraction for the analysis of short-chain chlorinated paraffins (SCCPs) in water samples. The extraction efficiency of five different commercially available fibres was evaluated and the 100-microm polydimethylsiloxane coating was the most suitable for the absorption of the SCCPs. Optimisation of several SPME parameters, such as extraction time and temperature, ionic strength and desorption time, was performed. Quality parameters were established using Milli-Q, tap water and river water. Linearity ranged between 0.06 and 6 microg l(-1) for spiked Milli-Q water and between 0.6 and 6 microg l(-1) for natural waters. The precision of the SPME-GC-ECD method for the three aqueous matrices was similar and gave relative standard deviations (RSD) between 12 and 14%. The limit of detection (LOD) was 0.02 microg l(-1) for Milli-Q water and 0.3 microg l(-1) for both tap water and river water. The optimised SPME-GC-ECD method was successfully applied to the determination of SCCPs in river water samples. PMID- 12564672 TI - In flow activation of diol-silica with cyanogen bromide and triethylamine for preparing high-performance affinity chromatographic columns. AB - A new coupling strategy using pre-packed diol-silica supports to obtain affinity columns for high-performance affinity chromatography (HPAC) is described. These columns were prepared by "in flow" activation in which solutions containing anhydrous solutions of CNBr and triethylamine are separately pumped to a mixer and then onto a pre-packed diol-silica column. Recycling the amino ligand to be coupled several times over the activated silica diol columns results in ligand immobilization. DNA (the Op 1 lac operator), 6-aminohexyl-Cibacron and a peptide (melittin) were all successfully "in flow" coupled to freshly activated columns. Methods for CNBr activation of pre-packed diol-silica column were developed for one, two or three pump HPLC systems. The supports were successfully used for the HPAC purification of a Lac repressor-beta-galactosidase fusion protein, alcohol dehydrogenase, and calmodulin. Columns prepared by in flow activation/coupling procedures were shown to be stable for at least 14 months. Also, in flow activated silica columns could be stored in anhydrous acetone for at least 3 months prior to coupling. Our experiments with these affinity ligand columns (DNA silica, aminohexyl-Cibacron F3GA-silica, and melittin-silica), suggests that this is a very successful coupling protocol for producing a variety of HPAC columns. PMID- 12564673 TI - Comparison of model monomeric and polymeric alkyl stationary phases on silica using sum-frequency spectroscopy. AB - Model monomeric and polymeric stationary phases were examined using sum-frequency spectroscopy and contact angle measurements in order to determine structural differences between the two stationary phases. Octadecyldimethylchlorosiloxane (ODMS) monolayers and ODMS/methyl siloxane (MS) mixed monolayers in contact with water and acetonitrile solvent were examined. The results showed that the alkyl chains in the ODMS and the ODMS/TMS monolayer are highly disordered and are weakly sensitive to the change in solvent environment. In order to determine whether the observed disorder in ODMS is due to steric effects of the silicon bonded methyl groups or the lack of crosslinking, "surface-bonded" ODS and "highly crosslinked" ODS were studied and compared with ODMS. Calculations using contact angle data on ODMS monolayers showed that water interacts 70% with methyl groups and 30% with methylene groups. PMID- 12564674 TI - Liquid chromatography of polyethylene glycol mono- and diesters: functional macromolecules or block copolymers? AB - The chromatographic behavior of polyoxyethylene-based polymers with adsorbing hydrophobic end-fragments was studied under two types of interaction conditions for the ethylene oxide (EO) component of such heteropolymers (which are either critical or of the size-exclusion type). In a theoretical part we assume a wide pore situation, where the molecules are smaller than pores, and consider models of two-component diblock and triblock copolymers having quite strongly adsorbing blocks. A new step is made to extend a theory from difunctional macromolecules with point-type end-groups to triblock copolymers. It is shown that A-B-A triblocks with adsorbing A-blocks and with a "critical" B-block behave in chromatography like difunctionals with effective end-group interactions. Another important finding is the existence of a second critical region, which was observed on most of the studied columns. This region can be used to separate di- and triblocks from each other. Additionally, the individual oligomers of triblocks can be separated to the baseline. Complete separations of both diblocks and triblocks in one single chromatogram can be achieved by using a step gradient between two types of studied condtions. PMID- 12564675 TI - Liquid chromatography with atmospheric pressure chemical ionization and electrospray ionization mass spectrometry of flavonoids with triple-quadrupole and ion-trap instruments. AB - With 15 flavonoids as test compounds, the analytical performance of four modes of LC-MS, multiple MS (MSn) and tandem MS operation (atmospheric pressure chemical ionization (APCI), electrospray ionization, positive and negative ionization) was compared for two mass spectrometers, a triple-quadrupole and an ion-trap instrument. Two organic modifiers, methanol and acetonitrile, and two buffers, ammonium acetate and ammonium formate, were used. In general, the use of APCI in the negative ion mode gave the best response, with the signal intensities and the mass-spectral characteristics not differing significantly between the two instruments. The best results were obtained when methanol-ammonium formate (pH 4.0) was used as LC eluent. Under optimum conditions full-scan limits of detection of 0.1-30 mg/l were achieved in the negative APCI mode. Here it needs to be emphasized that up to 2-order response differences were found both between analytes and between modes of ionization. This implies that one should be very cautious when interpreting data on the screening of real-life samples. The main fragmentations observed in the MSn spectra on the ion-trap, or the tandem MS spectra on the triple-quadrupole were generally the same. The advantage of the former approach is the added possibility to ascertain precursor-->product ion relationships. PMID- 12564676 TI - Ion trap tandem mass spectrometric identification of thiabendazole phototransformation products on titanium dioxide. AB - The purpose of this study is to artificially produce degradation intermediates of thiabendazole, which could be reasonably similar to those really present in the environment. The formation of by-products from thiabendazole transformation has been evaluated by adopting irradiated titanium dioxide as a photocatalyst. Several species more hydrophilic than the thiabendazole have been identified and characterized by HPLC-multiple MS. A pattern of reactions accounting for the observed intermediates is proposed. Two different parallel pathways are operating (and through these pathways the transformation of the molecule proceeds) leading to several intermediate compounds. The main steps involved are: (1) the hydroxylation of the molecule on the aromatic ring with the formation of a species having [M+H]+ 218; a further oxidation leads to the ring-opening and to the formation of aldehydic and alcoholic structures ([M+H]+ 270, 268 and 152); and (2) the cleavage of a C-C bond and the formation of a species having [M+H]+ 119. PMID- 12564677 TI - Practical implications of ionic strength effects on particle retention in thermal field-flow fractionation. AB - Modification of ionic strength of an aqueous or non-aqueous carrier solution can have profound effects on the particle retention behavior in thermal field-flow fractionation (ThFFF). These effects can be considered as either advantageous or not depending on the performance criteria under consideration. Aside from the general increase in retention time of particulate material (latexes and silica particles), our experiments indicate improvement in resolution with increases in electrolyte concentration. Absence of an electrolyte in the carrier solution causes deviations from the theoretically expected linear behavior between the retention parameter lambda (a measure of the extent of interaction between the applied field and the particle) and the reciprocal temperature drop across the channel walls. A negative interaction parameter delta(w), of about -0.170 was determined for 0.105- and 0.220-microm polystyrene (PS) latex particles suspended in either a 0.25 or a 1.0 mM TBAP-containing acetonitrile carrier and for 0.220 microm PS in 0.50 and 1.0 mM NaCl-containing aqueous medium. This work also demonstrates that optimum electrolyte concentrations can be chosen to achieve reasonable experimental run-times, good resolution separations, and shifts in the steric inversion points at lower field strengths, and that too high electrolyte concentrations can have deleterious effects such as band broadening and sample loss through adsorption to the channel accumulation surface. The advantages of using ionic strength rather than field strength to effect desired changes are lowered power consumption and possible application of ThFFF to high temperature labile samples. PMID- 12564678 TI - Determination of dithiocarbamate pesticides in occupational hygiene sampling devices using the isooctane method and comparison with an automatic thermal desorption (ATD) method. AB - Two new methods for the determination of dithiocarbamate pesticides in occupational hygiene sampling devices are described. Dithiocarbamate spiked occupational hygiene sampling devices, consisting of glass fibre (GF/A) filters, cotton pads, cotton gloves and disposable overalls, were reduced under acidic conditions and the CS2 evolved as a decomposition product was extracted into isooctane. The isooctane was then analysed using gas chromatography with mass spectrometry, for CS2, which provided a quantitative result for dithiocarbamates. Recoveries obtained were generally within a 70-110% range and reproducibilities better than 15% RSD were typically achieved. The method has been successfully applied to samples collected during occupational exposure surveys. A second method employing automatic thermal desorption-gas chromatography-mass spectrometry (ATD-GC-MS) has also been developed and applied to the direct analysis of GF/A (airborne) samples. The method relies on the thermal degradation of dithiocarbamates to release CS2, which is used to quantify the analytes. Thiram spiked GF/A filters gave an average recovery of 107% with an RSD of 4%. The performance of the two analytical methods were directly compared by analysing sub-portions of GF/A filters collected during a survey to evaluate occupational exposures to thiram during seed treatment operations. Both methods performed well for the analysis of airborne (GF/A) samples and produced results in good agreement. ATD-GC-MS is the preferred method for studies involving GF/A (airborne) samples only. Because of the wider applicability of the isooctane method for other sampling devices, it is the preferred choice when carrying out surveys which require a dermal as well as respirable exposure assessment. PMID- 12564679 TI - Evaluation of headspace solid-phase microextraction for the analysis of volatile carbonyl compounds in spirits and alcoholic beverages. AB - A method was developed for the determination of C1-C6 carbonyl compounds in alcoholic solutions using pentafluorobenzoxymation followed by headspace sampling solid-phase microextraction and subsequent analysis by GC with electron-capture detection. Experimental conditions-alcohol content, exposure time, temperature and sample agitation were optimised. In this method, a spirit or distilled alcoholic beverage is first adjusted to 20% (v/v) alcohol. Detection limits for particular aldehydes and ketone varied from 0.05 to 0.5 microg/l and relative standard deviation was between 2.3 and 20%. Generally, the method showed good linearity for the tested concentration range 8 microg/l-0.32 mg/l with regression coefficients ranging between 0.9434 and 0.9983. The method was applied to the analysis of real alcoholic beverages (vodkas). PMID- 12564680 TI - Protein proteolysis and the multi-dimensional electrochromatographic separation of histidine-containing peptide fragments on a chip. AB - This paper reports a system for three-dimensional electrochromatography in a chip format. The steps involved included trypsin digestion, copper(II)-immobilized metal affinity chromatography [Cu(II)-IMAC] selection of histidine-containing peptides, and reversed-phase capillary electrochromatography of the selected peptides. Trypsin digestion and affinity chromatography were achieved in particle based columns with a microfabricated frit whereas reversed-phase separations were executed on a column of collocated monolithic support structures. Column frits were designed to maintain constant cross sectional area and path length in all channels and to retain particles down to a size of 3 microm. Cu(II)-IMAC selection of histidine-containing peptides from standard peptide mixtures and protein digests followed by reversed-phase chromatography of the selected peptides was demonstrated in the electrochromatography mode. The possibility to run a comprehensive proteomic analysis by combining trypsin digestion, affinity selection, and a reversed-phase separation on chips was shown using fluorescein isothiocyanate-labeled bovine serum albumin as an example. PMID- 12564681 TI - Use of dynamically coated capillaries with added cyclodextrins for the analysis of opium using capillary electrophoresis. AB - A rapid, precise, accurate, and robust method using capillary electrophoresis (CE) with dynamically coated capillaries for the analysis of the major opium alkaloids in opium is presented. Dynamic coating of the capillary surface is accomplished using a commercially available reagent kit (polycation coating followed by polyanion coating). The addition of dual cyclodextrins (hydroxypropyl beta-cyclodextrin and dimethyl-beta-cyclodextrin) to the run buffer imparts excellent selectivity for the opium alkaloids. For the determination of morphine, papaverine, codeine, noscapine and thebaine in opium gum and opium latex samples (using tetracaine as an internal standard) good agreement with values obtained by gradient high-performance liquid chromatography is obtained. Compared to the latter technique, CE affords better resolution with significantly faster analysis time (12 min versus 29 min). Dynamically coated capillaries, which give rise to a relatively high and robust electroosmotic flow (EOF) at the background electrolyte pH of 2.5, allow for rapid analysis and excellent migration time and peak area precision (RSDs < or = 0.12% and < or = 1.2%, respectively). Reproducible separations (relative migration times) for over 500 samples have been obtained on a single capillary. The nature of the injection solvent, the injection time and the contents of the waste vials have a profound effect on the pressure injection precision of the relatively hydrophobic solutes. The CE conditions reported in this study are also applicable to the analysis of lysergic acid diethylamide (LSD) exhibits. PMID- 12564682 TI - Exploration of the electrophoretic behaviour of borane cluster anions and of the capability of capillary electrophoresis to separate them chirally. AB - Mobilities of investigated boron cluster compounds in 3-(N morpholino)propanesulfonic and phosphate buffers adjusted to pH 7 either with sodium hydroxide or with tris(hydroxymethyl)aminomethane depend on both buffer ions. The zone width and zone asymmetry, which are usually markedly higher than those of organic or common inorganic ions of comparable size, depend on the type of the borane cluster anion. Unusual shapes of zones of two investigated compounds have been found in tris phosphate buffer. Acetonitrile was superior to methanol as an organic additive to separation systems from the viewpoint of the zone symmetry and separation speed. Narrow trigonal zones, typical of organic ions non-interacting with the capillary wall, have been observed for some bridged sandwich cobalt complexes in run buffers with the addition of acetonitrile. The interaction of borane cluster anions with beta-cyclodextrin cavity is excessively strong in purely aqueous solutions. Methanol and acetonitrile, which generally weaken the interaction, sometimes affect the separation enantioselectivity of various compounds in different ways in addition to the weakening effect. Chiral discrimination was reached for all ten investigated anions, which belong to four different structural types of cluster boranes. Stability constants estimated for some analyte-beta-cyclodextrin complexes range between 100 and 1800 l/mol in acceptable separations. The relative difference of the constants was from 3 to 20%. PMID- 12564683 TI - Optimization and validation of a micellar electrokinetic chromatographic method for the analysis of several angiotensin-II-receptor antagonists. AB - We have optimized a micellar electrokinetic capillary chromatographic method for the separation of six angiotensin-II-receptor antagonists (ARA-IIs): candesartan, eprosartan mesylate, irbesartan, losartan potassium, telmisartan, and valsartan. A face-centred central composite design was applied to study the effect of the pH, the molarity of the running buffer, and the concentration of the micelle forming agent on the separation properties. A combination of the studied parameters permitted the separation of the six ARA-IIs, which was best carried out using a 55-mM sodium phosphate buffer solution (pH 6.5) containing 15 mM of sodium dodecyl sulfate. The same system can also be applied for the quantitative determination of these compounds, but only for the more stable ARA-IIs (candesartan, eprosartan mesylate, losartan potassium, and valsartan). Some system parameters (linearity, precision, and accuracy) were validated. PMID- 12564684 TI - Separation of andrographolide and neoandrographolide from the leaves of Andrographis paniculata using high-speed counter-current chromatography. AB - The bioactive diterpenes andrographolide and neoandrographolide from the leaves of Andrographis paniculata NEES (Acanthaceae) were successfully separated by counter-current chromatography. A single 280-min separation yielded 189 mg of 99.9% andrographolide and 9.5 mg of 98.5% neoandrographolide applying water methanol-ethyl acetate-n-hexane (2.5:2.5:4:1) solvent system. Structure confirmation was done by electrospray MS, one-dimensional NMR experiments, circular dichroism, optical rotation dispersion, and specific optical rotation [alpha]D. PMID- 12564685 TI - Comparison of separation conditions and ionization methods for the liquid chromatography-mass spectrometric determination of sulfonamides. AB - The effects of the type and concentration of buffer, composition of the mobile phase and the ionization mode, used for the separation and detection of sulfonamides with LC-MS, were studied. Five typical sulfonamides were selected as target compounds and beef meat was selected as a matrix sample. For the separation of sulfonamides, 0.05 M NH4Ac in 13-15% aqueous acetonitrile, APCl ionization was more effective than ESI with regard to separation efficiency and the detection sensitivity. PMID- 12564686 TI - Analysis by gas chromatography-mass spectrometry of the essential oils from the aerial parts of Rutheopsis herbanica (Bolle) Hans. & Kunk., gathered in Fuerteventura (Canary Islands). AB - The essential oil from the aerial parts of Rutheopsis herbanica (Bolle) Hans. & Kunk., growing in Fuerteventura, Canary Islands, Spain, was studied by gas chromatography and gas chromatography-mass spectrometry, and 42 constituents were identified. The major components were found to be alpha-pinene (29.4%), dillapiole (21.3%), limonene (14.1%), beta-pinene (13.2%) and myristicin (10.0%). As far as we know, this is the first report on the essential oil composition of this species. PMID- 12564687 TI - Preparation and application of a new modified liquid chromatographic chiral stationary phase based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid. AB - As an effort to improve the chiral recognition efficiency of a previously reported chiral stationary phase (CSP) based on (+)-(18-Crown-6)-2,3,11,12 tetracarboxylic acid, a new CSP was prepared by simply replacing the amide N-H hydrogens of the tethering groups of the old CSP with methyl groups. The new CSP was superior to the old one in the resolution of racemic primary amines. However, in the resolution of alpha-amino acids and amino alcohols, the new and the old CSPs were complementary with each other. The elution orders on the new CSP were sometimes opposite to those on the old one. Consequently, the chiral recognition mechanism on the new CSP was presumed to be different from that on the old one. The chiral recognition behavior of the new CSP were investigated with four selected analytes and found to be dependent to some extent on the content of organic and acidic modifiers in aqueous mobile phase and the column temperature. PMID- 12564688 TI - On-line solid-phase extraction of ceramides from yeast with ceramide III imprinted monolith. AB - A molecularly imprinted polymeric monolith (MIPM) was prepared by in situ polymerization using styrene, glycidyl methacrylate and methacrylic acid as monomers, divinylbenzene and triallyl isocyanurate as cross-linking agents, and ceramide III as print molecule. The texture, pore size distribution, mobile phase flow characteristic, and chromatographic performance of the MIPM and a control monolith synthesized without the print molecule were examined, respectively. The results showed that using ceramide III as print molecule significantly affected the pore structure and pore distribution of the monolith, and greatly improved the retention of ceramide III and its analogues used in cosmetics as well. The retention of ceramide III on the MIPM could be reduced by increasing the ratio of chloroform to hexane in eluting buffer. The workability of the MIPM was firstly demonstrated through the separation of a model lipid mixture containing ceramide III and ergosterol, the main sterol impurity in yeast lipid extracts. The application of the ceramide III imprinted monolith to the isolation of ceramides from yeast lipid extracts was attempted and resulted in a considerable enrichment of ceramides, as shown by FIIR analysis. This indicates the potential of ceramide III imprinted monolith synthesized in the present study in the on-line solid phase extraction of ceramides from yeast. PMID- 12564689 TI - Structural characterization by both positive and negative electrospray ion trap mass spectrometry of oligogalacturonates purified by high-performance anion exchange chromatography. AB - The off-line coupling of high-performance anion-exchange chromatography to electrospray ion trap mass spectrometry (ESI-IT-MS) is described. Two sets of isocratic conditions were optimised for the semi-preparative purification of oligogalacturonates of degree of polymerisation from 4 to 6 by monitoring eluates with either pulsed amperometric detection or evaporative light scattering detection in the presence of an online Dionex Carbohydrate Membrane Desalter (CMD). In these conditions, purified oligogalacturonate solutions were suitable, without further desalting steps, for infusion ESI-IT-MS experiments. This paper provides some interesting features of positive and negative ESI-IT-multiple MS (MSn) of these acidic oligosaccharides. The spectra acquired in both ion modes show characteristic fragments resulting from glycosidic bond and cross-ring cleavages. Under negative ionization conditions, the fragmentation of the singly charged [M-H]- ions, as well as the Ci-, and Zi-, fragment ions through sequential MSn experiments, was always dominated by product ions from C- and Z type glycosidic cleavages. All spectra also displayed 0.2 A-type cross-ring cleavage ions which carry linkage information. Collision-induced dissociation (CID) spectra of sodium-cationized species obtained under positive ionization conditions were more complex. Successive MSn experiments also led to the 0.2 A type cross-ring cleavage ions observed together with B- and Y-type ions. The presence of the 0.2 A ion series was related to Mr 60 (C2H4O2) losses. Combined with the absence of the Mr 30 (CH2O) and the Mr 90 (C3H6O3) ions, these ions were indicative of 1-4 type glycosidic linkage. PMID- 12564690 TI - Determination of estrogens and their conjugates in water using solid-phase extraction followed by liquid chromatography-tandem mass spectrometry. AB - An analytical procedure for the determination of steroid estrogens and their conjugates was developed and applied to aqueous environmental samples. The analytes of 15 compounds were solid-phase extracted and fractionated into two fractions: one containing unconjugated (free) steroids and the other containing conjugates (sulfates and glucuronides). Identification and quantification were carried out using liquid chromatography coupled with tandem mass spectrometry. The recoveries for each compound ranged from 57 to 116% and reproducibilities represented as RSD ranged from 2.9 to 17%. Some of the sulfates and free compounds were detected in environmental samples, whereas most of the conjugates were below the detection limits. PMID- 12564691 TI - Determination of carnitine and acylcarnitines in urine by high-performance liquid chromatography-electrospray ionization ion trap tandem mass spectrometry. AB - A high-performance liquid chromatography-mass spectrometry method has been developed for the simultaneous determination of native carnitine and eight acylcarnitines in urine. The procedure uses a solid-phase extraction on a cation exchange column and the separation is performed without derivatization within 17 min on a reversed-phase C8 column in the presence of a volatile ion-pairing reagent. The detector was an ion trap mass spectrometer and quantification was carried out in the MS-MS mode. Validation was done for aqueous standards at ranges between 0.75 and 200 micromol/l, depending on the compound. Carnitine was quantified in urine and comparison with a radioenzymatic assay gave a satisfactory correlation (R2 = 0.981). The assay could be successfully applied to the diagnostic of pathological acylcarnitines profile of metabolic disorders in urines of patients suffering from different organic acidurias. PMID- 12564692 TI - Enhancement of anion-exchange chromatography of DNA using compaction agents. AB - The use of adsorptive chromatography for preparative nucleic acid separations is often limited by low capacity. The possibility that the adsorbent surface area sterically accessible to nucleic acid molecules could be increased by reducing their radius of gyration with compaction agents has been investigated. The equilibrium adsorption capacity of Q Sepharose anion-exchange matrix for plasmid DNA at 600 mM NaCl was enhanced by up to ca. 40% in the presence of 2.5 mM spermine. In addition, compaction agent selectivity has been demonstrated. Spermine, for example, enhances the adsorption of both plasmid and genomic DNA, spermidine enhances binding only of plasmid, and hexamine cobalt enhances only the binding of genomic DNA. Compaction may be generally useful for enhancing adsorptive separations of nucleic acids. PMID- 12564693 TI - Micellar liquid chromatography determination of B vitamins with direct injection and ultraviolet absorbance detection. AB - A micellar reversed-phase liquid chromatographic procedure was developed for the control of five water-soluble vitamins, B (nicotinamide), B1 (thiamine), B2 (riboflavin), B6 (pyridoxine and pyridoxamine), in multivitamin pharmaceutical formulations (capsules, pills and syrups). Optimization procedure includes studies about the composition of the mobile phase (sodium dodecyl sulphate and the modifiers propanol, butanol or pentanol), flow-rate and temperature. Chromatographic analysis of all vitamins was carried out using a single mobile phase of 0.1 M SDS-4% (v/v) pentanol at pH 3, in a C18 column in isocratic mode, and UV-detection at 270, 290 and 325 nm. The flow-rates selected were 1.0 ml/min in the interval 0 to 6 min, and 2.0 ml/min until the end of the chromatogram and temperature was 45 degrees C. In the micellar liquid chromatographic system, the samples were injected without pretreatment, and the analysis time was below 12 min. Repeatabilities and intermediate precision were achieved according to ICH, and were below 5%. When the method is applied to real samples, the amount found with respect to the declared compositions were within the 91-105% range. These results were similar to those obtained with a conventional 60:40 (v/v) methanol water mixture for some of the vitamins, but with the advantage of use a single mobile phase for the analyses of the five vitamins, with direct injection of the samples and reduced toxicity, flammability, environmental impact and cost of the micellar-pentanol solutions. PMID- 12564694 TI - High-performance liquid chromatographic determination of polysorbate 80 in pharmaceutical suspensions. AB - A quick HPLC method is reported for the analysis of polysorbate 80 in pharmaceutical suspensions. A typical pharmaceutical suspension was mixed with dilute potassium hydroxide, and heated at 40 degrees C for 6 h. This procedure resulted in quantitative hydrolysis of polysorbate 80 to release oleic acid. A quick HPLC procedure was used to analyze the hydrolyzed samples without further sample treatment. Polysorbate 80 USP, treated in the same way as the pharmaceutical suspensions, was used as standard. Full validation tests were carried out and the validation studies demonstrated that this method is suitable for accurate and reproducible analysis of polysorbate 80 in pharmaceutical suspensions. PMID- 12564695 TI - Determination of 4-alkylphenols by novel derivatization and gas chromatography mass spectrometry. AB - A simple and sensitive method for the determination of alkylphenols in water samples has been developed using gas chromatography-mass spectrometry. Alkylphenols were determined after the extractive derivatization with pentafluoropyridine. The derivatization of alkylphenols efficiently proceeded to give the corresponding 4-tetrafluoropyridyl derivatives under the biphasic reaction system. The derivatization conditions including the phase-transfer catalyst, the amount of pentafluoropyridine, the reaction time, the concentration of NaOH and organic solvent were optimized. On the mass spectra of these derivatives, intense specific ion peaks were observed: m/z 256 for 4-n alkylphenols and m/z 284 for 4-tert.-alkylphenols. Calibration curves were linear in the range of 20-1000 ng/l (200-10,000 ng/l for nonylphenol), and the detection limits varied between 6.93 and 15.7 ng/l (85.2 ng/l for nonylphenol). The average recoveries of the alkylphenols in a fortified river water sample (100 ng/l except for nonylphenol: 1000 ng/l) ranged from 91.1 to 112%. The relative standard deviations were found to be between 5.6 and 16%. This method was successfully applied to the determination of alkylphenols in river water. PMID- 12564696 TI - Application of single-drop microextraction and comparison with solid-phase microextraction and solid-phase extraction for the determination of alpha- and beta-endosulfan in water samples by gas chromatography-electron-capture detection. AB - Water contamination due to the wide variety of pesticides used in agriculture practices is a global environmental pollution problem. The 98/83 European Directive requires the measurement of pesticides residues at a target concentration of 1.0 microg/l in surface water and 0.1 microg/l in drinking water. In order to reach the level of detection required, efficient extraction techniques are necessary. The application of a new extraction technique: single drop microextraction (SDME), followed by gas chromatography with electron-capture detection, was assessed for determining alpha-endosulfan and beta-endosulfan in water samples. Experimental parameters which control the performance of SDME, such as selection of microextraction solvent and internal standard, optimization of organic drop volume, effects of sample stirring, temperature and salt addition, and sorption time profiles were studied. Once SDME was optimized, analytical parameters such as linearity, precision, detection and quantitation limits, plus matrix effects were evaluated. The SDME method was compared with solid-phase microextraction and solid-phase extraction with the aim of selecting the most appropriate method for a certain application. PMID- 12564697 TI - Determination of pKa values of 2-amino-2-oxazolines by capillary electrophoresis. AB - The dissociation constants of new 2-amino-2-oxazolines were determined by capillary electrophoresis (CE) as a new technique. A method based on a linear model has been used in the CE determination. A series of eight 2-amino-2 oxazolines are investigated to determine their ionization constant. Among them, three new oxazolines synthesized are presented. The Ka values were obtained from the plots of reciprocal effective mobility against inverse concentrations of protons. The potentiometric method (PM) was performed as a comparative method. No significant differences were observed between the determined dissociation constants using both methods. Thus, the pKa values have been found to vary between 8.55 and 8.68. PMID- 12564698 TI - Chiral separation of amines with N-benzoxycarbonylglycyl-L-proline as selector in non-aqueous capillary electrophoresis using methanol and 1,2-dichloroethane in the background electrolyte. AB - N-Benzoxycarbonylglycyl-L-proline (L-ZGP) has been introduced as a chiral selector for enantioseparation of amines in non-aqueous capillary electrophoresis. Methanol mixed with different proportions of dichloromethane, 1,2-dichloroethane or 2-propanol containing L-ZGP and ammonium acetate was used as the background electrolyte. Enantioseparation of different types of pharmacologically active amines was performed, e.g. the local anaesthetic bupivacaine and the beta-adrenoceptor blocking agent pindolol. Addition of the solvents (dichloromethane, 1,2-dichloroethane or 2-propanol) gave an improved chiral separation partly due to a distinct decrease in the electroosmotic flow. The use of 1,2-dichloroethane in the background electrolyte gave higher precision in migration time (RSD 2.2%) compared to the systems containing dichloromethane. An enantiomeric separation of mepivacaine was performed within 72 s by use of short-end injection with an effective capillary length of 8.5 cm. PMID- 12564699 TI - Molecularly imprinted monolithic stationary phases for liquid chromatographic separation of enantiomers and diastereomers. AB - The method for preparation of molecularly imprinted monolithic stationary phase has been improved to achieve liquid chromatographic separation of enantiomers and diastereomers. By adopting low polar porogenic solvents of toluene and dodecanol and optimal polymerization conditions, the molecularly imprinted monolithic stationary phases with good flow-through properties and high resolution were prepared. Enantiomers of amino acid derivatives and diastereomers of cinchona alkaloids were completely resolved using the monolithic stationary phases. The influence of porogenic composition, monomer-template ratio and polymerization conditions on the chromatographic performance was investigated. Some chromatographic conditions such as the composition of the mobile phase and the temperature were characterized. Scanning electron microscopy showed that the molecularly imprinted monolithic stationary phase has a large through-pore structure to allow the mobile phase to flow through the column at very low backpressure. Accelerated separations of enantiomers and diastereomers were therefore achieved at elevated flow rates. Finally, the chiral recognition performance of the prepared stationary phase in aqueous media was investigated. Hydrophobic interaction, and ionic and/or hydrogen bonding interactions were proposed to be responsible for the recognition mechanism. PMID- 12564700 TI - Separation of negatively charged nonsteroidal anti-inflammatory drugs by reversed phase capillary electrochromatography. AB - Reversed-phase capillary electrochromatography in a 5-microm C18 fully packed capillary was employed to optimize the separation of negatively charged nonsteroidal anti-inflammatory drugs. The effect of the physico-chemical parameters and different analysis modes on the separation of 2-arylpropionic acids was studied and evaluated. The mobile phase composition, buffer type, concentration and pH differently influenced the peak efficiency and resolution, selectively modulating the analytes interaction with the stationary phase. The use of zwitterionic MES or acetate mobile phases strongly modulated the analytes migration order and peak efficiency. The optimum experimental conditions were found in MES buffer, pH 5.0, containing the 75% acetonitrile-methanol (1:1). All the analytes were baseline separated in a mixture in less than 13 min with peak efficiencies in the range of 78,500-84,200 N/m. Under these conditions the analytes were negatively charged and their effective electrophoretic mobilities played a role in the separation. The analysis of different pharmaceutical preparations containing anti-inflammatory drugs, e.g. drops and tablets, is also presented after a very simple sample pretreatment. PMID- 12564701 TI - Capillary electrophoretic determination of inorganic selenium species. AB - The performance of pyromellitic electrolyte for capillary zone electrophoresis of inorganic selenium species in the presence of selected common anions with indirect UV detection was investigated. The separation was achieved with pyromellitic electrolyte at pH 8.8 and hexamethonium hydroxide as the electroosmotic flow modifier. Obtained detection limits of 0.17 microg ml(-1) for Se(VI) and 0.29 microg ml(-1) for Se(IV) were improved by a factor of 5-7 in comparison with chromate electrolyte, which has been mainly employed for selenium analysis. Good resolution for nitrate-Se(VI) peaks were obtained. PMID- 12564702 TI - To thine own ethics code be true. PMID- 12564703 TI - Social work labor market: a first look. AB - Virtually no literature exists on the labor market for social workers. Fragmentary evidence suggests that this market is highly segmented by education; that many people who do not have social work degrees work in social work jobs, at least at the lower levels; that people who want to be social workers have such a strong commitment to the career of social work that relative wages make relatively little difference to their career decisions; and that employers often hire applicants without degrees for social work jobs. These factors tend to depress wages for social workers. Suggestions are made for a research and data collection program to make possible a definitive analysis of the social work labor market. PMID- 12564704 TI - So how far have we come? Pestilent and persistent gender gap in pay. AB - This article explores the issue of women's salaries in the human services within a comparative framework of many service occupations. An analysis of year-end 1998 data from the Bureau of Labor Statistics clearly demonstrates that salary disparities continue to exist between men and women. The author argues that these differences are based on continued patterns of discrimination, despite a plethora of policy initiatives dating back to the 1960s civil rights era to address gender discrimination in the workplace. Relevant policies are reviewed and assessed in terms of how far we have come in achieving pay equity between men and women. Several strategic directions to combat inequities are discussed, including public and professional education; individual, group, and professional advocacy; and targeted policy practice. Parallels are drawn between the gender discrimination experienced by social workers and client groups served. PMID- 12564705 TI - Managed care and the evolving role of the clinical social worker in mental health. AB - This article discusses developments for practitioners in mental health care with the advent of managed care. With managed care's influence and new cost containment strategies, the roles of clinical social workers, clinical psychologists, and psychiatrists are evolving, creating role conflict and competition. Along with changes in the utilization and definition of mental health practitioners' responsibilities, managed care has influenced issues such as private practice, mode of treatment, the use of outcome measurement and management, and the importance of case management. This article outlines the changes that have occurred and provides projections for the future. PMID- 12564706 TI - An in-law comes to stay: examination of interdisciplinary conflict in a school based health center. AB - Social workers often work in settings, such as hospitals, schools, prisons, nursing homes, and mental health facilities, where other professions exert a higher level of control because of their number, their historical precedence, or their professional status. The interdisciplinary conflicts that arise may be difficult to resolve. The organizational literature on causes of conflict and conflict resolution is briefly reviewed. A case study of a newly opened school based health center provides an opportunity to analyze conflicts between the school and health center personnel and demonstrates how they were finally resolved. PMID- 12564707 TI - From barriers to successful collaboration: public schools and child welfare working together. AB - Few mechanisms exist to support successful collaboration between public schools and child welfare agencies. One unfortunate consequence is that the children ostensibly being served by either system often end up receiving inadequate services from both systems. Focus groups were held with caseworkers, educators, and students to learn how the two systems can work more collaboratively. This article reports on the barriers and successful practices identified by the participants that affect the educational functioning of students living in foster care. The article concludes with the participants' recommendations for practices and policies to improve collaborative efforts between the two systems. PMID- 12564708 TI - Changing times: understanding social workers' motivation to be field instructors. AB - Renewing the profession of social work is historically dependent on social work practitioners volunteering to be field educators for students placed in their agencies. During the past decade, with massive funding cuts to social and health services and changes in professional practice, the availability of field instructors has been threatened. This article reports on social workers' motivations to become field instructors. The findings from qualitative interviews with 20 social workers randomly selected from a range of field agencies indicate that current organizational culture has a powerful influence on social workers' motivations to volunteer to become field instructors. The implications of this shift are discussed in relation to rejuvenating the profession of social work. Collaboration among organizations, professional associations, and schools of social work at the local and national level is critical. PMID- 12564709 TI - Facilitating writing by practitioners: survey of practitioners who have published. AB - The profession would benefit if practitioners viewed disseminating knowledge, including writing for publication, as part of professional practice. The literature has acknowledged the importance of practitioners' contributions to the profession's knowledge base, but little is known about what facilitates practitioners' writing for publication or what barriers they experience. A survey was mailed to practitioners who had published in social work peer-reviewed journals to obtain information about practitioners' experiences and to solicit their ideas on what the profession, human services agencies, and schools of social work could do to enhance the ability of practitioners to write for publication. Practitioners' recommendations for efforts that might increase writing for publication include continued collaborative efforts between academic researchers and practitioners, as well as suggestions for what social work educators and social services agencies can do to enhance the ability of practitioners to write for publication. PMID- 12564710 TI - Understanding adjudication: origins, targets, and outcomes of ethics complaints. AB - Various bodies and standards regulate social work practice. Social workers can be held accountable by licensure or credentialing boards, by civil and criminal laws, and by various professional organizations to which they belong. NASW considers complaints against its members for violations of the organization's Code of Ethics, yet the average member may know little about the nature and process of such actions. This article describes the professional review process and analyzes the results of a study of ethics cases filed with NASW from 1986 to 1997. This article also describes the parties involved in complaints and violations, the disposition of cases, and the corrective actions required of those who were found in violation of the Code. These research findings are compared with NASW membership data and the results of other studies on malpractice and ethics violations. PMID- 12564711 TI - Ethical dilemmas of reporting suspected elder abuse. AB - Elder abuse reporting laws exist in every state to protect elderly people being abused physically or emotionally, suffering from neglect, or experiencing financial exploitation. Support groups for caregivers of elderly individuals may help prevent elder abuse by reducing caregiver stress and linking caregivers to community services. Using case illustrations, the authors raise the dilemmas that group facilitators face when deciding whether to report a group member suspected of abusing his or her elder care recipient. The ethical concerns of reporting or withholding a report are discussed, and recommendations to assist facilitators in this complex decision process are presented. PMID- 12564712 TI - Value differences between social workers and members of the working and middle classes. AB - Although significant differences in values between social workers and clients are widely understood to affect the efficacy of service provision, no studies have sought to examine how the values affirmed by social workers may differ from those held by members of the working and middle classes. Therefore, this study examines the degree of value similarity between social workers and consumers. Based on "new-class" theory, two hypotheses are proposed. First, graduate social workers affirm value positions to the left of working- and middle-class clients. Second, bachelor's-level social workers affirm value positions in between those of graduate workers and clients. Both hypotheses were supported. The implications for the divergence in value frameworks for advocacy, practice, and education are discussed. PMID- 12564713 TI - Boundary issues in social work: managing dual relationships. AB - Social work literature clearly demonstrates that ethical issues related to boundaries are among the most problematic and challenging. Boundary issues involve circumstances in which social workers encounter actual or potential conflicts between their professional duties and their social, sexual, religious, or business relationships. This article provides an overview of boundary issues in social work (circumstances involving dual and multiple relationships); presents a conceptually based typology of boundary issues in the profession; and provides guidelines to help social workers manage the boundary issues and risks that arise in practice. PMID- 12564714 TI - Beyond individual rights and freedoms: metaethics in social work research. AB - Increasingly, social workers are called on to demonstrate the efficacy of their interventions and to contribute to knowledge building in the social sciences. Although social workers have a long tradition of practice ethics, less attention has been given to the unique dimensions of research ethics for social workers. A social work model of research ethics would consider how to balance highly valued ethical principles that are individually focused, such as self-determination and nonmalfeasance (the obligation to do no harm), with equally important values that have a collective focus, such as justice and beneficence (the obligation to bring about good). This article reviews current principles guiding research ethics, such as autonomy, beneficence, nonmalfeasance, and justice and provides an outline of the salient issues for social workers as they strive to address individual and collective interests in research endeavors, such as a greater emphasis on the social justice mission and the need to ensure that social justice objectives do not obscure individual rights and freedoms. The article concludes with preliminary recommendations for developing a social work perspective in research ethics. PMID- 12564715 TI - Competitive spillovers across non-profit and for-profit nursing homes. AB - The importance of non-profit institutions in the health care sector has generated a vast empirical literature examining quality differences between non-profit and for-profit nursing homes. Recent theoretical work has emphasized that much of this empirical literature is flawed in that previous studies rely solely on dummy variables to capture the effects of ownership rather than accounting for the share of non-profit nursing homes in the market. This analysis considers whether competitive spillovers from non-profits lead to higher quality in for-profit nursing homes. Using instrumental variables to account for the potential endogeneity of non-profit market share, this study finds that an increase in non profit market share improves for-profit and overall nursing home quality. These findings are consistent with the hypothesis that non-profits serve as a quality signal for uninformed nursing home consumers. PMID- 12564716 TI - Enrollee mix, treatment intensity, and cost in competing indemnity and HMO plans. AB - Why do indemnity insurance plans cost substantially more per capita--77% more in our study--than HMOs? We answer this question using data from a large organization's insurance pool, covering 215,000 lives. We decompose cost differences for eight major medical conditions into four sources: demographics, incidence within demographic groups, treatment intensity, and prices per service. Greater incidence of disease in the indemnity plan (both from demographics themselves and within demographic groups) and higher prices each explain nearly 50% of the difference. Contrary to conventional wisdom, indemnity plans do not have greater treatment intensity. PMID- 12564717 TI - A game theoretic model of the relationship between prices and waiting times. AB - ISSUE: The role of prices charged to patients as a policy instrument for altering waiting times. METHOD: A game theoretic model is used to examine the determinants of waiting times. RESULTS: Under certain circumstances an increase in the price charged to patients at some institutions will lower the waiting times at all institutions. CONCLUSIONS: The results provide a set of policy implications and testable predictions that arise under a certain set of assumptions. Alternative assumptions as well as the policy implications and testable predictions associated with them are also examined. PMID- 12564718 TI - Inequalities in self-reported health: validation of a new approach to measurement. AB - This paper assesses the internal validity of using the McMaster 'Health Utility Index Mark III' (HUI) to scale the responses on the typical self-assessed health (SAH) question, "how do you rate your health status in general?" It compares alternative procedures to impose cardinality on the ordinal responses. These include OLS, ordered probit and interval regression approaches. The cardinal measures of health are used to compute and to decompose concentration indices for income-related inequality in health. These results are validated by comparison with the individual variation in the 'benchmark' HUI responses obtained from the Canadian 'National Population Health Survey 1994-1995'. The interval regression approach, which exploits a mapping from the empirical distribution function (EDF) of HUI into SAH, outperforms the other approaches. In addition, we show how the method can be extended to allow for differences in SAH thresholds across different groups of people and to measuring and decomposing 'pure' health inequality. PMID- 12564719 TI - Prejudice, clinical uncertainty and stereotyping as sources of health disparities. AB - Disparities in health can result from the clinical encounter between a doctor and a patient. This paper studies three possible mechanisms: prejudice of doctors in the form of being less willing to interact with members of minority groups, clinical uncertainty associated with doctors' differential interpretation of symptoms from minority patients or from doctor's distinct priors across races, and stereotypes doctors hold about health-related behavior of minority patients. Within a unified conceptual framework, we show how all three can lead to disparities in health and health services use. We also show that the effect of social policy depends critically on the underlying cause of disparities. PMID- 12564720 TI - Alcohol dependence and the price of alcoholic beverages. AB - This study estimates the impact of the price of alcoholic beverages on latent dimensions of current alcohol dependence and abuse. A three-part econometric model is used to estimate the impact of price on three latent dimensions (factors). For heavier drinking, the estimated price elasticity is -1.325 (P = 0.027); for physical and other consequences of drinking, -1.895 (P = 0.003); for increased salience of drinking, -0.411 (P = 0.339). For a single latent factor characterized simply as dependence/abuse, estimated price elasticity is -1.487 (P = 0.012). These results suggest that higher prices for alcohol reduce important dimensions of current alcohol dependence and abuse. PMID- 12564721 TI - Editorial: Small intestinal bacterial overgrowth in dogs--less common than you think? PMID- 12564722 TI - Chronic intestinal inflammation and intestinal disease in dogs. AB - Normal individuals maintain tolerance to the endogenous bacterial flora residing within their alimentary tract, a phenomenon mediated by the gastrointestinal lymphoid tissue. Loss of this tolerance is a key factor in the development of chronic intestinal inflammation. Manifestations of such uncontrolled inflammation in humans include inflammatory bowel disease and celiac disease. Dogs may similarly be affected, and although the etiopathogenesis is likely similar, the lesions differ. This review includes discussion of the factors involved in breakdown of mucosal tolerance, the immunologic basis of canine enteropathies, and the use of novel immunotherapies for these diseases. PMID- 12564723 TI - Increased mean arterial pressure and aldosterone-to-renin ratio in Persian cats with polycystic kidney disease. AB - Polycystic kidney disease (PKD) in Persian cats has been increasingly reported and compared to human autosomal dominant polycystic kidney disease (ADPKD) in the last decade. In cats, however, few studies have dealt with the occurrence and hormonal determinants of hypertension, one of the most common extrarenal manifestations of ADPKD in humans. The purpose of this study was to compare Persian cats >4 years old with PKD to unaffected control cats with regard to blood pressure (BP), plasma renin activity (PRA), serum aldosterone concentration, plasma atrial natriuretic peptide (ANP) concentration, and aldosterone-to-renin ratio (ARR). Three gender- and age-matched groups were studied, each consisting of 7 cats: (1) a control group without cysts, (2) a group with mild PKD, and (3) a group with severe PKD (multiple cysts and renal enlargement). Mild renal insufficiency was found in only 1 of 14 cats with PKD. Cats with PKD had a higher mean arterial pressure (P = .04) and more often had a high ARR (P = .047) than did control cats. Tendencies toward higher diastolic and systolic arterial pressures (DAPs and SAPs, respectively) and lower PRAs were observed in cats with PKD compared to controls (.05 < P < or = .1). No significant differences were found between the groups in serum aldosterone and plasma ANP concentrations. None of the cats had echocardiographic evidence of cardiac hypertrophy. In conclusion, cats with PKD had a minor increase in mean arterial pressure compared to control cats, and half of the cats had a high ARR. PMID- 12564724 TI - Congenital dilatation of the bile ducts (Caroli's disease) in young dogs. AB - We describe 8 young dogs with congenital dilatation of the intra- and extrahepatic bile ducts and diffuse cystic kidney disease, compatible with Caroli's disease in humans. The dogs were referred between 1980 and 2000 because of chronic disease at an age of 6 months to 3 years. These dogs included 3 Collies, 2 Frisian Stabyhouns, 2 Jack Russell Terriers, and 1 mixed-breed dog. The most common signs were vomiting (6/6), polyuria and polydipsia (4/6), and anorexia (4/6). Ascites was a common finding (4/6). Clinicopathologic abnormalities were available for 6 dogs. All had increased plasma alkaline phosphatase activity and fasting bile acids: increased alanine aminotransferase activity and urea and creatinine concentrations were present in 50% of dogs. Ultrasound examination of the liver showed severely dilated bile ducts without evidence of obstruction, and calcification in all cases but 1. Postmortem examination revealed severe dilatation of the larger intra- and extrahepatic bile ducts. The common bile duct and gall bladder were normal, and the bile system was patent. The ducts contained a clear viscid fluid often with calcified material. Microscopically, marked portal fibrosis was present, often with abnormally structured dilated bile ducts lined with columnar or cuboid epithelium and regularly small calcifications. The lesion was complicated by ascending cholangitis in 1 dog. The kidneys showed marked cortical and medullary fibrosis with a diffuse radial cystic pattern; only slight renal fibrosis was found in the oldest dog. Seven dogs were euthanized without treatment; the oldest dog was alive and well 5 months after diagnosis and was maintained on a protein restricted diet. PMID- 12564726 TI - Evaluation of the effects of clomipramine on canine thyroid function tests. AB - To evaluate the effect of long-term clomipramine administration on the hypothalamic-pituitary-thyroid axis in healthy dogs, 14 healthy adult dogs were enrolled in a prospective study. Clomipramine (3 mg/kg PO q12h) was administered to all dogs beginning on day 0, and continued for 112 days. Serum total thyroxine (T4), free thyroxine (fT4), 3,5,3'-triiodothyronine (T3), 3,3',5' triiodothyronine (reverse T3; rT3), and thyroid-stimulating hormone (TSH) were measured on days 0, 7, 28, 42, 56, and 112. Thyrotropin-releasing hormone (TRH) response tests were performed concurrently. Significant decreases were noted in serum T4, f4, and rT3 concentrations beginning on day 28 through the end of the study period. The lowest mean (+/-SEM) concentrations of T4 (26 +/- 1.2 to 17 +/- 0.5 nmol/L) and rT3 (1.21 +/- 0.13 to 0.83 +/- 0.08 nmol/L) occurred at day 112, whereas the lowest mean fT4 (29 +/- 2.4 to 18 +/- 1.7 pmol/L) was found on day 56 of clomipramine treatment. The effect of treatment over time on serum T3 concentration also was significant, but the deviation in T3 from baseline was variable. No significant effect of clomipramine treatment was noted on either pre or post-TRH TSH concentrations. The 35 and 38% decreases in serum T4 and fT4 concentrations, respectively, during clomipramine administration may lead to a misdiagnosis of hypothyroidism. Although no evidence of hypothyroidism was noted in this study population, subclinical hypothyroidism may have occurred. A longer duration of treatment might further suppress thyroid function, and concurrent illness or other drug administration might exacerbate clomipramine's effects. PMID- 12564725 TI - Comparison of direct and indirect tests for small intestinal bacterial overgrowth and antibiotic-responsive diarrhea in dogs. AB - Controversy exists over the diagnosis of idiopathic small intestinal bacterial overgrowth (SIBO) in dogs and some clinicians use the term antibiotic-responsive diarrhea (ARD) in preference. However, whether such terms are interchangeable is not clear. To examine the relationship between duodenal bacterial numbers and a clinical response to antibiotics, SIBO and ARD were defined by nonoverlapping criteria. Quantitative duodenal juice bacteriology and indirect serum biochemical tests were used to assess small intestinal bacterial populations in 30 dogs with gastrointestinal disorders, including 9 with ARD. Serum total unconjugated bile acid (TUBA) concentrations were measured in all dogs, serum folate and cobalamin concentrations were measured in 29 of 30 dogs, and quantitative culture of duodenal juice was performed in 22 of 30 dogs. Serum TUBA concentrations also were measured in samples from 38 control dogs. Twenty of 22 affected (clinical) dogs in which quantitative bacteriology was performed were classified as having SIBO (>10(5) colony-forming units of total bacteria per milliliter of duodenal juice), but bacterial numbers did not differ significantly between dogs with ARD and dogs with other disorders. Increased folate (19/29), decreased cobalamin (16/ 29), or a combination (9/29) were common, but increased TUBA concentrations were documented in only 5 of 30 clinical dogs. Again, no significant differences were observed between dogs with ARD and those with other disorders, and a similar proportion (5/38) of controls had abnormally high TUBA concentrations. Finally, no significant differences were noted when duodenal bacteriology and TUBA concentrations were assessed before and during antibiotic therapy. These results question the utility of quantitative duodenal juice bacteriology and indirect biochemical marker tests for SIBO in the investigation of canine gastrointestinal disorders. PMID- 12564727 TI - Congenital hypothyroidism with goiter in toy fox terriers. AB - Congenital hypothyroidism with goiter was observed to segregate as a simple autosomal recessive trait in Toy Fox Terriers (TFTs). Neonatal affected pups exhibited inactivity, abnormal hair coat, stenotic ear canals, and delayed eye opening. Palpable ventrolateral cervical swellings were evident by 1 week of age. Serum thyroid hormone and thyroid-stimulating hormone concentrations were low and high, respectively. Histologic examination of the cervical masses disclosed cuboidal to columnar follicular epithelial cell hyperplasia with widely varying follicular size, shape, and amount of colloid. Oral thyroid hormone replacement therapy restored near-normal growth and development. At 8 weeks of age, radioiodine uptake and perchlorate discharge testing indicated an iodine organification defect. Biochemical analysis of thyroid tissue from affected dogs demonstrated enzymatic iodine oxidation deficiency and lack of sodium dodecyl sulfate-resistant thyroglobulin dimers, suggesting thyroid peroxidase deficiency. A nonsense mutation in the thyroid peroxidase gene of affected dogs was discovered and demonstrated to segregate with the disease. A DNA-based carrier test was developed and currently is used by TFT breeders to prevent this disorder. PMID- 12564728 TI - A comparison of dobutamine infusion to exercise as a cardiac stress test in healthy horses. AB - This study was done to determine whether administration of dobutamine would produce echocardiographic and electrocardiographic alterations comparable to those induced by treadmill exercise in healthy horses. Fourteen horses received maximal treadmill exercise and, separately, intravenous dobutamine infusion up to a maximum rate of 50 microg/kg/min. Ten of the 14 horses were euthanized, and the myocardial tissues were examined grossly and histopathologically. No significant differences were found in the chronotropic effects of dobutamine and exercise (P = .905). Dobutamine induced greater interventricular septal thickening during systole (dobutamine = 4.78 cm, exercise = 4.03 cm; P = .004). and greater left ventricular diameters during diastole (dobutamine = 9.73 cm, exercise = 9.26 cm; P = .037), than did exercise treatment. Horses exhibited transient signs of sweating and restlessness during infusion of moderate to maximum doses of dobutamine. Ventricular ectopy seen in 11 of 14 horses was attributed to the arrhythmogenic properties of dobutamine, as well as to increased vagal tone present at low dobutamine doses. Myocardial lesions characteristic of catecholamine myotoxicity were present in 2 of the 10 horses examined. Although dobutamine induces chronotropic and inotropic changes similar to those induced by exercise, the use of high-dose dobutamine as a cardiac stressor in horses cannot be advocated because of potential development of arrhythmias or myotoxicity. PMID- 12564729 TI - Comparative diagnostic test characteristics of oscillometric and Doppler ultrasonographic methods in the detection of systolic hypertension in dogs. AB - Comparison of test characteristics allows a clinician to choose the optimal diagnostic test method for an individual patient. This study assessed the comparative test characteristics of noninvasive (NI) blood pressure measurement methods (oscillometric and Doppler) and used this information to develop optimal cutoff values for diagnosis of systolic hypertension in dogs by these NI methods. Simultaneous NI (oscillometric or Doppler methods) and invasive (arterial puncture [AP]) systolic blood pressure (SBP) measurements were obtained prospectively from normal dogs and dogs suspected of having systemic hypertension based on clinical signs. Oscillometric SBP readings were obtained from the distal hind limb (Osc-L, n = 54) or the proximal tail (T. n = 27). Doppler BP measurements were obtained using a forelimb cuff (n = 57). AP-SBP was categorized as hypertensive if > or = 160 mmHg, and sensitivity (Se). specificity (Sp), and likelihood ratios (LR) were calculated for diagnostic cutoff values ranging from 130 to 220 mmHg. Receiver operator characteristic (ROC) curves were analyzed to determine optimal cutoff values for diagnosis of AP-SBP > or = 160 mmHg. Optimal NI SBP cutoff values considered to reflect AP values > or = 160 mmHg were: Osc-L = 160 mmHg (Se: 65%, Sp: 85%. LR = 4.33: 1), Osc-T = 150 mmHg (Se: 84%, Sp: 75%, LR = 3.36: 1), and Doppler = 160 mmHg (Se: 71%, PMID- 12564730 TI - Arterial thromboembolism in cats: acute crisis in 127 cases (1992-2001) and long term management with low-dose aspirin in 24 cases. AB - Records of 127 cats with arterial thromboembolism (ATE) were reviewed. Abyssinian, Birman, Ragdoll, and male cats were overrepresented. Tachypnea (91%), hypothermia (66%), and absent limb motor function (66%) were common. Of 90 cats with diagnostics performed, underlying diseases were hyperthyroidism (12), cardiomyopathy (dilated [8], unclassified [33], hypertrophic obstructive [5], hypertrophic [19]), neoplasia (6), other (4), and none (3). Common abnormalities were left atrial enlargement (93%), congestive heart failure (CHF, 44%), and arrhythmias (44%). Of cats without CHF, 89% were tachypneic. Common biochemical abnormalities were hyperglycemia, azotemia, and abnormally high serum concentrations of muscle enzymes. Of 87 cats treated for acute limb ATE, 39 (45%) survived to be discharged. Significant differences were found between survivors and nonsurvivors for temperature (P < .00001), heart rate (P = .038), serum phosphorus concentration (P = .024), motor function (P = .008), and number of limbs affected (P = .001). No significant difference was found between survivors and nonsurvivors when compared by age, respiratory rate, other biochemical analytes, or concurrent CHE A logistic regression model based on rectal temperature predicted a 50% probability of survival at 98.9 degrees F (37.2 degrees C). Median survival time (MST) for discharged cats was 117 days. Eleven cats had ATE recurrences, and 5 cats developed limb problems. Cats with CHF (MST: 77 days) had significantly shorter survival than cats without CHF (MST: 223 days; P = .016). No significant difference was found in survival or recurrence rate between cats receiving high-dose aspirin (> or = 40 mg/cat q72h) and cats receiving low-dose aspirin (5 mg/cat q72h). Adverse effects were less frequent and milder for the lower dosage. PMID- 12564731 TI - Regurgitant fraction measured by using the proximal isovelocity surface area method in dogs with chronic myxomatous mitral valve disease. AB - Previous studies have demonstrated that regurgitant fraction can be measured by using the proximal isovelocity surface area (PISA) method. For this study, we utilized this Doppler echocardiographic method to estimate the magnitude of mitral regurgitation in dogs with myxomatous mitral valve disease. Seventeen older, small dogs with chronic mitral regurgitation and no to mild myocardial failure were studied. A blinded observer judged the clinical severity of mitral regurgitation to be mild, moderate, or severe by using echocardiographic assessment of left heart size. The regurgitant fraction was calculated by using the PISA method and spectral Doppler echocardiography. The regurgitant fraction was compared to the clinical assessment of severity for each dog and to calculations of left atrial size. Five dogs had clinically mild mitral regurgitation. Four of these dogs had a regurgitant fraction between 22 and 41%, whereas 1 had a regurgitant fraction of 73%. The 3 dogs with clinical evidence of moderate mitral regurgitation had a regurgitant fraction of 46-65%. All 9 dogs with clinically severe mitral regurgitation had a regurgitant fraction greater than 75% (78-88%). The regurgitant fraction was statistically different between each group (P < .001). A good but curvilinear relationship was found between left atrial size and regurgitant fraction (r2 = 0.72). In this study, dogs with clinically severe mitral regurgitation consistently had hemodynamically severe regurgitation (regurgitant fraction > 75%), whereas dogs with clinically mild to moderate disease had lesser degrees of regurgitation. Good correlation was found between regurgitant fraction and left atrial size. We conclude that the major determinant of left atrial size and disease severity in dogs with mitrial regurgitation is the degree of mitral regurgitation. PMID- 12564732 TI - Spontaneous feline hypertension: clinical and echocardiographic abnormalities, and survival rate. AB - Systemic hypertension was diagnosed in 58 of 188 untreated cats referred for evaluation of suspected hypertension-associated ocular, neurologic. cardiorespiratory, and urinary disease, or diseases frequently associated with hypertension (hyperthyroidism and chronic renal failure). Hypertensive cats were significantly older than normotensive subjects (13.0 +/- 3.5 years versus 9.6 +/- 5.0 years; P < .01), and had a greater prevalence of retinal lesions (48 versus 3%; P < .001), gallop rhythm (16 versus 0%; P < .001), and polyuria-polydipsia (53 versus 29%: P < .01). Blood pressure was significantly higher (P < .001) in cats with retinopathies (262 +/- 34 mm Hg) than in other hypertensive animals (221 +/- 34 mm Hg). Hypertensive cats had a thicker interventricular septum (5.8 +/- 1.7 versus 3.7 +/- 0.64 mm; P < .001) and left ventricular free wall (6.2 +/- 1.6 versus 4.1 +/- 0.51 mm; P < .001) and a reduced diastolic left ventricular internal diameter (13.5 +/- 3.2 versus 15.8 +/- 0.72 mm; P < .001) than control cats. Left ventricular geometry was abnormal in 33 of 39 hypertensive subjects. No significant difference was found in age or blood pressure at the initial visit between cats that died or survived over a 9-month period after initial diagnosis of hypertension. Mean survival times were not significantly different between hypertensive cats with normal and abnormal left ventricular patterns. Further prospective studies are needed to clearly identify the factors involved in survival time in hypertensive cats. PMID- 12564733 TI - Treatment of dogs with oral melanoma by hypofractionated radiation therapy and platinum-based chemotherapy (1987-1997). AB - This retrospective study in 39 dogs with incompletely resected oral melanoma examined the efficacy of hypofractionated radiation therapy and platinum containing chemotherapy. All dogs were completely staged, with the majority of dogs classified as stage 1. Dogs received 6 weekly fractions of 6-gray (Gy) megavoltage irradiation with a cobalt-60 unit or a 4-MeV (megaelectron volts) linear accelerator. Dogs received cisplatin (10-30 mg/m2 IV) or carboplatin (90 mg/m2 IV) chemotherapy 60 minutes before radiation delivery. Durations of local control, metastasis-free survival time, and overall survival time were recorded. By the Kaplan-Meier method, 15% of the dogs had local recurrence within a median time of 139 days. Fifty-one percent of the dogs developed metastatic disease within a median time of 311 days (range, 24-2, 163 days). Median survival time for all 39 dogs was 363 days. The combined use of chemotherapy and radiation therapy in this protocol provided local control consistent with previous studies. Low-dose chemotherapy was used with the intent of enhancing radiation therapy for the local control of an incompletely excised tumor. Survival times were longer than previously reported for dogs with oral malignant melanoma. Additional studies are required to determine whether these results were due to the effects of chemotherapy on microscopic disease or the enhanced local control provided by chemoradiation therapy. PMID- 12564734 TI - Influence of host factors on survival in dogs with malignant mammary gland tumors. AB - The purpose of our study was to determine if specific host factors, such as age at diagnosis, obesity, and hormone status, influence the prognosis of canine mammary gland carcinomas and to confirm if previously reported risk factors (ie, histologic subtype, tumor size, and World Health Organization [WHO] stage) were important in a large series of affected dogs. Ninety-nine female dogs with mammary gland carcinomas, no previous therapy, an excisional biopsy, and known cause of death were studied. No significant association with survival was noted for age at diagnosis (chronologic or physiologic), obesity, or hormone status (ie, spayed versus intact, regardless of time of being spayed). Of the tumor factors analyzed, the histologic subtype anaplastic carcinoma (P = .02), WHO stage I (P = .01), evidence of metastasis at the time of diagnosis (P = .004), and tumor size of 3 cm or smaller (P = .005) all significantly influenced survival. Dogs that were classified as having tumor-related mortality had a shorter postoperative survival compared to dogs that died of other causes (14 months versus 23 months; P = .03). In conclusion, histologic subtype, WHO stage, and tumor size remain important prognostic factors in canine mammary gland tumors. Further study of other prognostic factors is needed to determine which tumors are adequately addressed with local therapy only and which dogs may require adjuvant treatment with chemotherapy. PMID- 12564735 TI - Gastrointestinal leiomyosarcoma in 14 dogs. AB - A retrospective review over a 5-year period was performed to identify clinical characteristic features and survival in 14 dogs with intestinal leiomyosarcoma after surgery. Anemia, hypoglycemia, polyuria, and polydipsia were identified as common clinical signs in these dogs. Dogs that survived the immediate postoperative period had a median survival of 21.3 months, (range 0.1-72.5 months). The 1- and 2-year survival rates were 75 and 66%, respectively. Dogs with intestinal leiomyosarcoma that survive the immediate postoperative period can experience long-term survival after surgical excision. Furthermore, dogs with histologically documented metastasis at the time of surgery also appear to have a long survival, with a mean survival of 21.7 months (range 4.2-41.5 months). PMID- 12564736 TI - Oleander toxicosis in a donkey. PMID- 12564737 TI - Emphysematous gastritis associated with Clostridium septicum in a horse. PMID- 12564738 TI - Management of pneumothorax in cattle by continuous-flow evacuation. AB - Pneumothorax in cattle can develop subsequent to acute or chronic pulmonary disease, and if unresolved may lead to respiratory distress and death due to hypoxia and compression and collapse of cardiac and thoracic great vessels. Therapeutic evacuation of free air within the pleural space can provide acute relief and improve chances of survival. This article descibes the adaptation and use of a continuous flow evacuation device to resolve pneumothorax in 3 cattle with pneumothorax associated with infectious lower airway disease. PMID- 12564739 TI - Pharmacoeconomics and the "4th hurdle": drug reimbursement policies in Central and Eastern Europe. PMID- 12564740 TI - Therapeutic drug monitoring of clozapine and relapse--a retrospective study of routine clinical data. AB - OBJECTIVE: Therapeutic drug monitoring (TDM) of the atypical antipsychotic drug clozapine is recommended. Clinical studies have indicated a therapeutic window for clozapine serum levels in schizophrenic and schizo-affective patients during acute treatment, i.e. for patients who do not respond to treatment with typical antipsychotics. However, despite the frequent use of clozapine also in maintenance treatment, very few data are available showing the relationship between serum levels of clozapine and the prevention of relapse. Thus, the primary objective of the study was to investigate the relationship between serum levels ofclozapine and relapse during maintenance treatment. METHODS: A retrospective study of routine TDM-data was conducted. Samples obtained on an acute treatment ward from patients with < 4 days hospitalization (recent admissions) were regarded as samples associated with relapse. Samples which can be attributed to an intoxication were identified as described in the TDM-form. The serum level of clozapine, as well as age, gender, smoking habits, concurrent drugs, psychiatric diagnosis and dose of clozapine were evaluated. Data analysis was performed on individual samples and, alternatively, on multiple samples from a single patient which are summarized according to a typical clinical situation. RESULTS: 404 serum levels were measured in 86 patients. After exclusion of patients receiving acute treatment, 65 relevant clinical situations were identified in 50 patients: 12 relapses, 8 intoxications (a total of 20 situations with poor outcome) and 45 situations involving patients with good maintenance outcome. Samples involving relapse had serum levels of 198 +/- 211 ng/ml (10 624), intoxications had serum levels of 1,969 +/- 705 ng/ml (900-2,900) and those with good outcome had serum levels of 384 +/- 255 ng/ml (56-1,028) (mean +/- SD (range)). By means of sensitivity of receiver operating characteristic curves (ROC) a lower limit of the therapeutic window can be estimated at about 50-250 ng/ml and an upper limit at about 745-1,050 ng/ml. The frequencies of good and poor outcome were significantly different within and outside these ranges, e.g. chi2 = 11.8 and p < 0.001 for 250 to 745 ng/ml. Comparison of only good outcome and relapse provided a significant difference in the serum level of clozapine (Student's t-test p = 0.024). However, 67% of relapses were predicted in a model of logistic regression only if the variables serum level and concurrent treatment with other psychotropic drugs were included simultaneously as independent variables. Neither variable was able to predict relapse if used as a single variable in separate models. Finally, it was found that serum levels of clozapine were increased in women, in aged patients and in nonsmokers. CONCLUSIONS: It is tentatively concluded that serum levels of clozapine < 50 ng/ml are related to relapse irrespective of concurrent psychotropic drugs. In cases where there are no concurrent psychotropic drugs, serum levels of clozapine < 250 ng/ml are associated with relapse. The risk of relapse is low for serum levels of clozapine > 250 ng/ml irrespective of concurrent psychotropic drugs. The risk of intoxication is increased with serum levels > 750 ng/ml. The TDM of clozapine is recommended during maintenance treatment. PMID- 12564741 TI - Differential therapy of constipation--a review. AB - Constipation is a common condition occurring with increasing frequency in advanced age. As a symptom, it is not always dealt with directly by the physician, but is often left to the care of nurses. Many patients rely on self medication. Constipation not only interferes with the quality of life, but often has the rank of a syndrome requiring competent medical intervention. This is of clinical importance, because a thorough understanding of the pathophysiology of constipation enables the clinician to identify the potential causes and, if necessary, initiate a differentiated therapy with the aid of only simple additional investigations. This review outlines a clinical approach including medical history, spectrum of causes, radiopaque pellets method to differentiate between slow transit constipation and defecation disturbances for the differential diagnosis of constipation. The mechanisms of action of laxatives (anti-absorptive, secretagogue, osmotic, filling and swelling agents) are further components and important for the individual therapy. Based on this fundamental information, a differentiated therapy is possible in each specific case such as coprostasis. Chronic symptomatic constipation reduces the quality of life and should be evaluated by physicians. If situations such as drug-induced constipation or hypothyroidism which have to be treated causally can be ruled out, laxative treatment according to the clinical picture, mode of action of drug used and side effects of the laxative can be initiated. Laxative abuse due to chronic constipation is rare and almost always associated with psychosomatic psychiatric disorders. PMID- 12564742 TI - Comparison of talinolol and atenolol effects on blood pressure in relation to lipid and glucose metabolic parameters. Results from the TALIP study. AB - AIMS: The primary objective of this double-blind, randomized, parallel-group study was to compare the influence ofthe selective beta1-receptor antagonists talinolol (100 mg) and atenolol (50 mg) on the lipid metabolism in hyperlipemic patients with mild to moderate hypertension after 12 weeks of treatment. As a secondary endpoint, the influence of the drug on blood pressure, pulse rate as well as glucose metabolism were examined. Furthermore, pharmacokinetic parameters were assessed. PATIENTS: Of the 198 patients recruited for the study, 166 were randomized to receive atenolol (n = 83) or talinolol (n = 83) for up to 12 weeks, 149 patients received the study medication for up to 48 weeks under double-blind conditions. RESULTS: There was no difference between the antihypertensive effect of both beta1-selective antagonists in patients with mild to moderate hypertension. No clinically relevant differences between the 2 drugs were observed for LDL cholesterol, HDL cholesterol, total cholesterol and triglycerides in the rather low doses given. However, there was evidence for a decrease in LDL cholesterol following treatment with talinolol, but not following treatment with atenolol, in patients with the highest initial blood pressure and in those with normalized blood pressure after 12 weeks of treatment. Parameters of glucose metabolism were not adversely affected by both drugs. Stable pharmacokinetics were observed over the 12-week administration, and steady state conditions were achieved after a 1-week treatment with both active compounds in the target population. Data indicate that once-a-day dosing can be performed with less fluctuation between peak and trough for talinolol in comparison to atenolol. Both treatments were well tolerated. PMID- 12564743 TI - Quercetin inhibits the sulfation of r(-)-apomorphine in human brain. AB - The first aim of this investigation was to study the sulfation of R(-) apomorphine in human brain. The second aim was to investigate the inhibition of R(-)-apomorphine sulfation by quercetin in human brain. R(-)-apomorphine is hereafter referred to as apomorphine. Apomorphine sulfation was measured in 5 brain specimens; 3 derived from the frontal cortex and 2 derived from the temporal cortex. The rate of apomorphine sulfation was 5.6 +/- 4.3 pmol/min/mg. The activities of SULT1A1 and SULT1A3, which were also measured in these samples, were 11 +/- 9.1 and 2.6 +/- 1.7 pmol/min/mg, respectively. The rate of apomorphine sulfation correlated with the activity of SULT1A1 (r = 0.989; p = 0.002) and SULT1A3 (r = 0.973; p = 0.005). Apomorphine sulfotransferase followed Michaelis-Menten kinetics, the Km (mean +/- SD) and Vmax values (mean +/- SD) of which, measured in 5 brain samples, were 32 +/- 7.3 microM and 8.9 +/- 7.9 pmol/min/mg, respectively. Quercetin was a potent inhibitor of apomorphine sulfation with an IC50 value, measured in 5 brain samples, of 16 +/- 2.3 nM. The inhibition mechanism of quercetin using apomorphine sulfation in 5 brain samples was mixed, non-competitive with a Ki and Kies (mean +/- SD) of 16 +/- 4.1 and 87 +/- 37 nM, respectively (p = 0.008). The intrinsic clearance value of apomorphine (mean +/- SD) was 247 +/- 170 ml/min/mg(-1) and was decreased to 100 +/- 85 ml/min/mg(-1) (p < 0.01) in the presence of 25 nM quercetin. In conclusion, apomorphine is sulfated in human brain. Sulfation might reduce the level of apomorphine in human brain and be a factor limiting the effect of this drug. Quercetin is a potent inhibitor of apomorphine sulfation and may inhibit the sulfation of apomorphine in human brain in vivo. PMID- 12564744 TI - Trends in ophthalmic antimicrobial utilization pattern in Bahrain between 1993 and 2000: a resurgence of chloramphenicol? AB - OBJECTIVES: The occurrence of aplastic anemia following topical administration of ophthalmic chloramphenicol is controversial and debated internationally. We have determined the influence of such debate on the utilization of ophthalmic chloramphenicol in Bahrain, through studying the utilization patterns of ophthalmic antimicrobial preparations by the Ministry of Health, with an emphasis on chloramphenicol, between 1993 and 2000. Cost-implications of these patterns are examined. MATERIAL AND METHODS: Information on the annual purchase of ophthalmic antimicrobial drug preparations and their unit price was obtained from the Directorate of Materials Management, Ministry of Health, and analyzed. RESULTS: In 1993, the 3 most commonly purchased ophthalmic antibacterial preparations were oxytetracycline 1% eye ointment (40.1%); sulfacetamide 10% and 20% eye drops (25.3%); and chloramphenicol 0.5% eye drops and 1% eye ointment (10.8%). In 2000, oxytetracycline remained the most frequently purchased preparation (33%), followed by chloramphenicol (21.2%). Between 1993 and 1999, chloramphenicol purchases fluctuated between 10% to 16.4% with a remarkable increase to 21.2%, in 2000. Chloramphenicol accounted for 8.6% and 15.1% of cost of total ophthalmic preparations purchased in 1993 and 2000, respectively. CONCLUSION: Despite continued concerns of potential risks of ophthalmic chloramphenicol, this preparation is extensively utilized in Bahrain. We are of the opinion that for minor infections, chloramphenicol ophthalmic preparations should be replaced by safer alternatives. Further, we recommend that their use be reserved for ocular infections that are resistant to other antimicrobials, and that ophthalmologists, at the secondary care level, should supervise such treatment. PMID- 12564745 TI - Comparative bioavailability of two novel coenzyme Q10 preparations in humans. AB - OBJECTIVE: To determine the absorptive properties of 2 novel coenzyme Q10 preparations, a fast-melting tablet and an effervescent tablet, compared with currently available formulations. MATERIALS AND METHODS: In the first trial, the absorptive properties of 4 different coenzyme Q10 preparations (fast-melting, effervescent, soft gelatin, and powder-filled hard shell) were studied in a randomized, single-dose, crossover study. Twenty-four male subjects were given a 60 mg dose of coenzyme Q10 and plasma coenzyme Q10 was measured over the next 12 hours. Pharmacokinetic properties including area under the curve (AUC), maximum plasma concentration (Cmax), time to maximum plasma concentration (Tmax) and elimination half-life (t 1/2) were measured. In a separate single-dose study, the absorptive characteristics of a different coenzyme Q10 soft gel (Q-Gel) were studied in 6 male subjects. RESULTS: Area under the curve (microg/ml x h) for the fast-melting and effervescent formulations, while marginally greater, was not significantly different when compared to the soft gelatin and powder-filled preparations, 5.4 +/- 1.04 (110%) and 5.5 +/- 0.589 (112%) versus 5.0 +/- 0.859 (102%) and 4.9 +/- 0.812 (100%), respectively. Cmax for the 2 novel formulations was also not statistically different from the soft gelatin or powder-filled preparations, 0.87 +/- 0.14 and 0.86 +/- 0.074 versus 0.70 +/- 0.010 and 0.81 +/- 0.159 (microg/ml). Tmax however, was significantly shorter for the fast-melting and effervescent formulations compared with the soft gel and powder-filled forms, 1.3 +/- 0.348 and 2.0 +/- 0.552 versus 3.7 +/- 0.702 and 4.1 +/- 0.993 (h), respectively. The results of the second trial were similar to those of the powder filled and soft gel formulations from the first study. CONCLUSIONS: The novel fast-melting and effervescent formulations provide a more rapid delivery of CoQ10 to the blood while exhibiting a similar AUC compared with current formulations. The potential clinical significance of this finding should be further evaluated. PMID- 12564746 TI - The nonuse of psychological research at two federal agencies. AB - In 1994 the Government Accounting Office (GAO) issued a report critical of some features of the proposal review processes at the National Science Foundation and the National Institutes of Health. I provide two examples of procedures the agencies could have adopted to address the GAO's criticisms. I also relate the history of the two agencies' reluctance to use the psychological research literature to guide them as their new review procedures were instituted. Finally, I enumerate possible reasons for the agencies' decision not to follow or even test suggestions based on the judgment and decision-making research literature. PMID- 12564747 TI - Emotional facilitation of sensory processing in the visual cortex. AB - A key function of emotion is the preparation for action. However, organization of successful behavioral strategies depends on efficient stimulus encoding. The present study tested the hypothesis that perceptual encoding in the visual cortex is modulated by the emotional significance of visual stimuli. Event-related brain potentials were measured while subjects viewed pleasant, neutral, and unpleasant pictures. Early selective encoding of pleasant and unpleasant images was associated with a posterior negativity, indicating primary sources of activation in the visual cortex. The study also replicated previous findings in that affective cues also elicited enlarged late positive potentials, indexing increased stimulus relevance at higher-order stages of stimulus processing. These results support the hypothesis that sensory encoding of affective stimuli is facilitated implicitly by natural selective attention. Thus, the affect system not only modulates motor output (i.e., favoring approach or avoidance dispositions), but already operates at an early level of sensory encoding. PMID- 12564748 TI - On wildebeests and humans: the preferential detection of negative stimuli. AB - On the basis of a functional perspective, we hypothesized that negative stimuli are detected faster than positive stimuli. In Experiment 1, participants were subliminally presented with positive and negative words or with no words at all. After each presentation, participants were asked whether they had seen a word. They detected negative words more accurately than positive words. In Experiment 2, participants were subliminally presented with negative or positive words. After each presentation, they were asked whether the presented word was positive or negative. Negative words were correctly categorized more often than positive words. Experiment 3 showed that although participants correctly categorized negative words more often than positive words. they could not guess the meaning of the words better than would be expected by chance. The results are discussed against the background of recent findings on basic affective processes. PMID- 12564749 TI - Long-term inhibition of return of attention. AB - During search of the environment, the inhibition of the return (IOR) of attention to already-examined information ensures that the target will ultimately be detected. Until now, inhibition was assumed to support search of information during one processing episode. However, in some situations search may have to be completed long after it was begun. We therefore propose that inhibition can be associated with an episode encoded into memory such that later retrieval reinstates inhibitory processing and encourages examination of new information. In two experiments in which attention was drawn to face stimuli with an exogenous cue, we demonstrated for the first time the existence of long-term IOR. Interestingly. this was the case only for faces in the left visual field, perhaps because more efficient processing of faces in the right hemisphere than the left hemisphere results in richer, more retrievable memory representations. PMID- 12564750 TI - Time course of perceptual grouping by color. AB - Does perceptual grouping operate early or late in visual processing? One position is that the elements in perceptual layouts are grouped early in vision, by properties of the retinal image, before perceptual constancies have been determined. A second position is that perceptual grouping operates on a postconstancy representation, one that is available only after stereoscopic depth perception, lightness constancy, and amodal completion have occurred. The present experiments indicate that grouping can operate on both a preconstancy representation and a postconstancy representation. Perceptual grouping was based on retinal color similarity at short exposure durations and based on surface color similarity at long durations. These results permit an integration of the preconstancy and postconstancy positions with regard to grouping by color. PMID- 12564751 TI - Critical evidence: a test of the critical-period hypothesis for second-language acquisition. AB - The critical-period hypothesis for second-language acquisition was tested on data from the 1990 U.S. Census using responses from 2.3 million immigrants with Spanish or Chinese language backgrounds. The analyses tested a key prediction of the hypothesis, namely, that the line regressing second-language attainment on age of immigration would be markedly different on either side of the critical-age point. Predictions tested were that there would be a difference in slope, a difference in the mean while controlling for slope, or both. The results showed large linear effects for level of education and for age of immigration, but a negligible amount of additional variance was accounted for when the parameters for difference in slope and difference in means were estimated. Thus, the pattern of decline in second-language acquisition failed to produce the discontinuity that is an essential hallmark of a critical period. PMID- 12564752 TI - Spontaneous attention to word content versus emotional tone: differences among three cultures. AB - A Stroop interference task was used to test the hypothesis that people in different cultures are differentially attuned to verbal content vis-a-vis vocal tone in comprehending emotional words. In Study 1, Americans showed greater difficulty ignoring verbal content than ignoring vocal tone (which reveals an attentional bias for verbal content); but Japanese showed greater difficulty ignoring vocal tone than ignoring verbal content (which reveals a bias for vocal tone). In Study 2, Tagalog-English bilinguals in the Philippines showed an attentional bias for vocal tone regardless of the language used, suggesting that the effect is largely cultural rather than linguistic. Implications for culture and-cognition research are discussed. PMID- 12564753 TI - Electrophysiological responses to errors and feedback in the process of action regulation. AB - The anterior cingulate cortex (ACC) is believed to be involved in the executive control of actions, such as in monitoring conflicting response demands, detecting errors, and evaluating the emotional significance of events. In this study, participants performed a task in which evaluative feedback was delayed, so that it was irrelevant to immediate response control but retained its emotional value as a performance indicator. We found that a medial frontal feedback-related negativity similar to the error-related negativity (ERN) tracked affective response to the feedback and predicted subsequent performance. Source analysis of the feedback-related negativity and ERN revealed a common dorsomedial ACC source and a rostromedial ACC source specific to the ERN. The oscillatory nature of these sources provides further evidence that the ERN reflects ongoing theta activity generated in the mediofrontal regions. These results suggest that action regulation by the cingulate gyrus may require the entrainment of multiple structures of the Papez corticolimbic circuit. PMID- 12564754 TI - New objects, not new features, trigger the attentional blink. AB - When two different targets must be selected from a rapid serial visual presentation (RSVP) of images, perception of the second target will be markedly reduced if it is presented within about a half second of the first. Known as the attentional blink (AB), this effect reflects temporal limitations in attentional processes enabling awareness of image representations. I tested whether these limitations occur at an object or feature level of processing by presenting (in RSVP) multiple images of the same (old) object depicted in different orientations. Targets were defined by new features added either to this or to a new object. When the first target feature appeared on the old object, no AB effects were found even when the second target was a new object. When a new object carried the first target feature, an AB effect was found even when the second target feature appeared on the same "new" object. The AB appears to reflect limitations in the creation of new object representations, rather than temporal limitations of awareness per se. PMID- 12564755 TI - The secret life of pronouns: flexibility in writing style and physical health. AB - Numerous disclosure studies have demonstrated that individuals randomly assigned to write about emotional topics evidence improved physical health compared with those who write about superficial topics. The writing samples from three previously published studies of 74 first-year students, 50 upper-division students, and 59 maximum-security prisoners were reanalyzed using Latent Semantic Analysis (LSA) to explore possible relationships of writing content and style to changes in frequency of physician visits following the disclosure intervention. LSA revealed that flexibility in the use of common words-particularly personal pronouns--when writing about traumatic memories was related to positive health outcomes. The findings point to the importance of the role of discussing the self and social relationships in writing and, at the same time, to the remarkable potential of techniques such as LSA. PMID- 12564756 TI - Serial expertise of rhesus macaques. AB - Here we describe the development of serial expertise in 4 experimentally naive rhesus monkeys that learned, by trial and error, the correct order in which to respond to 3-, 4-, and 7-item lists of arbitrarily selected photographs. The probabilities of guessing the correct sequence on 3-, 4-, and 7-item lists were, respectively, 1/6, 1/24, and 1/5,040. Each monkey became progressively more efficient at determining the correct order in which to respond on new lists. During subsequent testing, the subjects were presented with all possible pairs of the 28 items used to construct the four 7-item lists (excluding pairs of items that occupied the same ordinal position in different lists). Subjects responded to pairs from different lists in the correct order 91% of the time on the first trials on which these pairs were presented. These features of subjects' performance, which cannot be attributed to procedural memory, satisfy two criteria of declarative memory: rapid acquisition of new knowledge and flexible application of existing knowledge to a new problem. PMID- 12564757 TI - High-fidelity perceptual long-term memory revisited--and confirmed. AB - Experiments on short-term perceptual memory for elemental visual attributes, such as contrast, motion, orientation, and spatial frequency, have relied on a delayed discrimination technique in which the subject compares two stimuli presented at different points in time and memory is indexed by discrimination thresholds measured for the different time intervals between reference and test. In a variant of this procedure, used in experiments on long-term memory, the presentation of a single reference is followed by a memory test that combines two alternative forced-choice decisions with the method of constant stimuli With this procedure, it is impossible to distinguish the effects of criterion-setting processes and memory on performance, but this confound can be eliminated by testing many subjects and having each subject make a single decision. The resulting "group thresholds" are stable across time intervals of 24 hr, confirming previous findings of high-fidelity storage in the long-term memory range. PMID- 12564758 TI - Absolut memory distortions: alcohol placebos influence the misinformation effect. AB - Can the simple suggestion that you have consumed alcohol affect your memory for an event? Alcohol placebos affect social behaviors but not nonsocial ones, and have not previously been shown to affect memory. We investigated the effect of alcohol placebos using materials that revealed both the social and the nonsocial influences of memory Subjects drank plain tonic water, but half were told it was a vodka and tonic; then all subjects took part in an eyewitness memory experiment. Subjects who were told they drank alcohol were more swayed by misleading postevent information than were those who were told they drank tonic water, and were also more confident about the accuracy of their responses. Our results show that the mere suggestion of alcohol consumption may make subjects more susceptible to misleading information and inappropriately confident. These results also provide additional confirmation that eyewitness memory is influenced by both nonsocial and social factors. PMID- 12564759 TI - "Blaming the victim" under memory load. AB - When presented with negative outcomes, people often engage in counterfactual thinking imagining various ways that events might have been different. This appears to be a spontaneous behavior, with considerable adaptive value. Nevertheless, counterfactual thinking may also engender systematic biases in various judgment tasks, such as allocating blame for a mishap, or deciding on the appropriate compensation to a victim. Thus, counterfactuals sometimes require thought suppression or discounting, potentially resource-demanding tasks. In this study. participants made mock-jury decisions about control and counterfactual versions of simple stories. The judgments of two groups of participants, differing in their respective levels of working memory capacity, were compared. In addition, all participants held memory loads during various stages of the primary task. Lower-span individuals were especially susceptible to bias associated with the counterfactual manipulation, but only when holding memory loads during judgment. The results suggest that counterfactual thoughts arise automatically, and may later require effortful, capacity-demanding suppression. PMID- 12564760 TI - Phonological influences on lexical (mis)selection. AB - Speakers produce words to convey meaning, but does meaning alone determine which words they say? We report three experiments that show independent semantic and phonological influences converging to determine word selection. Speakers named pictures (e.g., of a priest) following visually presented cloze sentences that primed either semantic competitors of the target object name ("The woman went to the convent to become a..."), homophones of the competitors ("I thought that there would still be some cookies left, but there were..."), or matched unrelated control object names. Primed semantic competitors (nun) were produced instead of picture names more often than primed unrelated control object names, showing the well-documented influence of semantic similarity on lexical selection. Surprisingly, primed homophone competitors (none) also substituted for picture names more often than control object names even though they only sounded like competitors. Thus, independent semantic and phonological influences can converge to affect word selection. PMID- 12564761 TI - The constitutive and inducible expression of Nurr1, a key regulator of dopaminergic neuronal differentiation, in human neural and non-neural cell lines. AB - Nur-related factor 1 (Nurr1), nerve growth factor-induced gene B (NGFI-B) and neuron-derived orphan receptor-1 (NOR-1) constitute the orphan nuclear receptor subfamily of transcription factors. Previous studies showed that midbrain dopaminergic neuronal precursor cells failed to differentiate in Nurr1-deficient mice. To investigate a role of Nurr1 in human neuronal function, Nurr1 mRNA expression was studied in human neural cell lines by RT-PCR and northern blot analysis. Nurr1, NGFI-B and NOR-1 mRNA were coexpressed in all human neural and nonneural cell lines under the serum-containing culture condition, except for SK N-SH neuroblastoma, in which Nurr1 mRNA was undetectable. The levels of Nurr1, NGFI-B and NOR-1 mRNA were elevated markedly in NTera2 teratocarcinoma-derived neurons (NTera2-N), a model of differentiated human neurons, following a 1.5 or 3 h-exposure to 1 mM dibutyryl cyclic AMP or 100 nm phorbol 12-myristate 13 acetate. NGFI-B mRNA levels were also elevated in NTera2-N cells by exposure to 100 ng/mL brain-derived neurotrophic factor (BDNF). To identify Nurr1-target genes, the mRNA expression of 27 genes potentially involved in dopaminergic neuronal differentiation and survival, including BDNF, glia-derived neurotrophic factor, their receptors, tyrosine hydroxylase and alpha-synuclein, were studied in HEK293 cells following overexpression of Nurr1. None of these genes examined, however, showed significant changes. These results indicate that Nurr1, NGFI-B and NOR-1 mRNA are expressed constitutively in various human neural and non neural cell lines under the serum-containing culture condition, and their levels are up-regulated in human neurons by activation of protein kinase A or protein kinase C pathway, although putative coactivators expressed in dopaminergic neuronal precursor cells might be required for efficient transcriptional activation of Nurr1-target genes. PMID- 12564762 TI - The reaction of glial progenitor cells in remyelination following ethidium bromide-induced demyelination in the mouse spinal cord. AB - The present study investigated how glial progenitor cells participated in the process of remyelination following ethidium bromide (EBr)-induced demyelination in the adult mouse spinal cord. In situ hybridization techniques for detecting mRNA for platelet-derived growth factor alpha receptor (PDGFalphaR) and proteolipid protein (PLP) were employed to identify glial progenitor cells and mature oligodendrocytes, respectively. During the demyelination stage and early stage of remyelination, large cells strongly expressing PDGFalphaR mRNA were observed in the border of the demyelinating lesion, and with immunohistochemistry they exhibited positive labeling of the astrocytic marker glial fibrillary acidic protein (GFAP). Other glial progenitor cells expressing PDGFalphaR mRNA proliferated around the lesion during the demyelination stage. During the remyelination stage, some PDGFalphaR mRNA-positive cells partly expressed mRNA for PLP in the periphery of the demyelinating lesion. These results suggest that PDGFalphaR mRNA-positive glial progenitor cells may give rise to both astrocytes and oligodendrocytes, which participate in remyelination following demyelination. PMID- 12564763 TI - Neuropathological investigation of cerebral white matter lesions caused by closed head injury. AB - In order to ascertain whether there is widespread axonal disruption of cerebral white matter in the so-called 'diffuse axonal injury' (DAI), a type of closed head injury, proposed by Adams et al. the author investigated his own cases clinicopathologically. Twenty-six male autopsied cases of head injury, aged between 19 and 84, 15 of which had sustained road traffic accidents, were examined; the others were due to falling from heights and so on. The study group all belonged to non-missile head injuries and included 12 cases of diffuse brain injury, as well as 14 cases of focal brain injury, according to the classification of Gennarelli et al. The survival time ranged from 2 h to 21 years. Formalin-fixed brains were cut coronally so as to make paraffin-embedded hemispheric sections. Then these sections were stained conventionally (HE, Bodian, Kluver-Barrera and Holzer) and immunohistochemically (GFAP) to assess axonal decrease, myelin pallor and gliosis by the use of light microscopy. In the 13 chronic cases that died more than 1 month after the accidents, the intensities of gliosis, myelin pallor and axonal decrease tended to correlate with each other. In the 13 acute cases who died less than 1 month after their accident, the degree of axonal decrease in white matter seemed to correlate with the severity of myelin pallor. Regardless of types of trauma, however, axonal retraction balls, the so-called hallmark of DAI, were found only with myelin pallor suggesting the presence of brain swelling after the injury. Therefore these findings indicate that it may be difficult to accept the notion of DAI, that is, the presence of axonal retraction balls without brain swelling. In addition, diffuse vascular injury (2 cases) as well as rarefaction of subcortical white matter (6 cases) were presented and their pathogenesis individually discussed based on a literature review. PMID- 12564764 TI - Atypical teratoid/rhabdoid tumor of the central nervous system: clinico pathological study. AB - Atypical teratoid/rhabdoid tumor (AT/RT) is a new entity among malignant pediatric brain tumors. This study was performed to investigate the clinicopathologic and cytogenetic features of the tumor. Five cases from a series of the brain tumors were studied. Clinical features of the patients were assessed with age, sex, location of the tumors, treatments and survival periods. Histopathologic features were analyzed by routine HE stain and immunohistochemical stains for epithelial membrane antigen, cytokeratin, vimentin, smooth muscle actin, desmin, GFAP, S-100 protein, neurofilament protein, synaptophysin and alpha-feto protein. Cytogenetic studies for karyotype analysis and fluorescent in situ hybridization were available in three cases. Mean age of patients was 5.6 years, and maximal survival periods were less than 13 months despite surgical and radiation therapy. The tumors were located in infratentorial and supratentorial areas. Histopathologically, the tumors were chiefly composed of rhabdoid cells, modified rhabdoid cells and undifferentiated small cells, mixed with epithelial, mesenchymal, and neural tumor-like areas. These polyphenotypic features of the tumor cells were supported by diverse immunoreaction. Monosomy of chromosome 22 was demonstrated in two cases of the tumor. These results suggest that AT/RT may be a unique clinicopathologic entity, and histopathologic diagnosis of it should be made judiciously by differentiating other polymorphous tumors of the brain. PMID- 12564765 TI - Changes in expression of p38 mitogen-activated protein kinase in the dorsal motor nucleus of the vagus nerve and hypoglossal nucleus after axotomy in adult rats. AB - Mitogen-activated protein (MAP) kinase cascades are activated in response to various extracellular stimuli. P38 MAP kinase is one of the MAP kinase family and is activated by proinflammatory cytokines and environmental stresses. Activating transcription factor-2 (ATF-2) is one of the targets for p38 MAP kinase. To obtain information on the role of the p38 MAP kinase in the neurons and glial cells after axotomy, we investigated changes of expression of p38 MAP kinase, MAP kinase kinase (MKK) 3, MKK4, MKK6 and ATF-2 in the dorsal motor nucleus of the vagus nerve and the hypoglossal nucleus following axotomy in rats using in situ hybridization and immunohistochemical techniques. Expression of p38 MAP kinase mRNA was observed in the neurons in control rats and showed no remarkable changes after axotomy in both nuclei. On the other hand, expression of p38 MAP kinase mRNA was observed in the perineuronal microglias after axotomy. The expression of p38 MAP kinase, activated p38 MAP kinase, MKK3 and ATF-2 were immunohistochemically observed in neurons of control rats in both nuclei. After axotomy, the expression of p38 MAP kinase, active and inactive, and ATF-2 in neurons were reduced in both nuclei, while expression of mRNA of p38 MAP kinase showed no reduction in neurons. These findings indicate that p38 MAP kinase is functionally regulated not by synthesis but by phosphorylation and regulates the activation of ATF-2 in neurons, and this cascade plays some role in retrograde neuronal reactions. Moreover, perineuronal microglial cells showed strong expression of p38 MAP kinase, active and inactive, after axotomy in both nuclei. These findings suggest that p38 MAP kinase is related to microglial cell reactions after axotomy. PMID- 12564766 TI - FKBP12 immunoreactivity in the human spinal cord of motor neuron disease patients. AB - We investigated the FKBP12 immunoreactivity in the spinal cord of neurological controls and motor neuron disease (MND) patients. In the neurological controls, the spinal neurons were markedly stained with antihuman FKBP12 (N-19 and C-19) antibodies. FKBP12 immunoreactivity was associated with lipofuscin in formalin fixed paraffin-embedded samples. In an electron microscopic view, the 10-nm colloidal gold particles labeled by the anti-FKBP12 (N-19) antibody were present on the lipofuscin of the spinal anterior horn neurons. In the MND cases, atrophic neurons with an abundance of lipofuscin granules in the anterior horns of the spinal cord were mildly stained with the anti-FKBP12 (N-19 and C-19) antibodies. Normal-appearing neurons, inclusion-laden neurons and chromatolytic neurons of MND cases were weak or negatively stained with anti-FKBP12 (N-19) antibodies. These findings suggest that FKBP12 (N-19) may decrease in the early stages of degeneration in MND. Complexes of FKBP12 and ligands were reported to have neuroprotective and/or neuroregenerative properties. It is speculated that the decrease in FKBP12 (N-19) plays some causative role in the development of neurodegeneration in MND. Further investigation of FKBP12 and ligands may help elucidate the pathogenesis of MND. PMID- 12564767 TI - Ischemic rat brain extracts induce human marrow stromal cell growth factor production. AB - Intravenous administration of human bone marrow stromal cells (hMSCs) after middle cerebral artery occlusion (MCAo) in rats provides functional benefit. We tested the hypothesis that these functional benefits are derived in part from hMSC production of growth and trophic factors. Quantitative sandwich enzyme linked immunosorbent assay (ELISA) of hMSCs cultured with normal and MCAo brain extracts were performed. hMSCs cultured in supernatant derived from ischemic brain extracts increased production of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF). These neurotrophins and angiogenic growth factors increased in a post-ischemia time-dependent manner. The hMSC capacity to increase expression of growth and trophic factors may be the key to the benefit provided by transplanted hMSCs in the ischemic brain. PMID- 12564768 TI - Neuropathology of experimental autoimmune encephalomyelitis modified by retroviral infection. AB - The A8 virus is a molecular clone of the neuropathogenic FrC6 virus derived from the Friend murine leukemia virus (F-MuLV). To elucidate the effects of A8 virus infection on immune-mediated diseases in the central nervous system, we investigated the development of acute and monophasic experimental autoimmune encephalomyelitis (EAE) in A8 virus-infected Lewis rats. In EAE rats after A8 virus infection (A8-EAE), many inflammatory cells were found in the gray matter including the frontal lobe, where almost no inflammatory cells were found in rats with EAE alone. The modified distribution of inflammatory cells was not dependent on the ages of A8 virus-infected rats, although the frequency of the modified distribution was reduced in older rats. The chimeric virus Rec2, which contains the pol and env genes of 57 virus on the background of A8 and does not induce spongiform degeneration in the CNS, caused the same distributional modification of inflammatory cells in the rats with EAE as in A8-EAE rats. Furthermore, the incidence and intensity of spongiform degeneration, thymoma and splenomegaly caused by A8 virus were reduced by the induction of EAE. PMID- 12564769 TI - Gliofibrous nodule in the cerebello-medullary fissure. AB - An extra-axial nodule in the cerebello-medullary fissure is described, occurring in a 27-year-old-woman. MRI and CT scans revealed the lesion was a non-enhanced round mass, which was associated with mild atrophy of the surrounding cerebellum, but with no perifocal edema. In the surgical observation, the mass was white, elastic and hard, well demarcated and localized in the cerebello-medullary fissure. Histologically, the lesion was composed of astrocytes and collagen producing fibroblasts with no anaplasia. These findings suggested that the lesion was hamartomatous, but not neoplastic. This type of gliofibrous nodule has not been previously reported. PMID- 12564770 TI - Co-existence of nemaline and cytoplasmic bodies in muscle of an infant with nemaline myopathy. AB - A sporadic case of congenital myopathy had severe muscle weakness of neonatal onset. Nemaline and cytoplasmic bodies were detected in muscle biopsies taken at 4 months of age. These findings were consistent with a diagnosis of nemaline myopathy (severe neonatal form). The simultaneous and abundant presence of these two types of sarcoplasmic inclusion has been found in only a few cases. However, these cases suggest that the sarcoplasmic inclusions may be formed, at least partially, by common mechanisms. PMID- 12564771 TI - Clinicopathological studies of peripheral neuropathy in Churg-Strauss syndrome. AB - Clinicopathological studies were performed on the visceral organs and the sural nerve of a male patient with Churg-Strauss syndrome (CSS) in order to understand the mechanisms of peripheral nervous system damage. A 67-year-old man, with a 2 year history of bronchial asthma, developed acutely painful paraplegia and dyspnea. Laboratory data showed a leukocytosis, an elevated serum creatinine kinase (CK) and marked eosionophilia. Autoantibodies including p- and c-ANCA were negative. Electrophysiological studies revealed a severe sensory-motor neuropathy of multiple mononeuritis type. Steroid pulse therapy performed a day after biopsy of skin, muscle and sural nerve was effective in resolving his respiratory and neurological dysfunction but a perforation of an intestinal ulcer occurred which required surgical intervention. In the biopsied sural nerve and the surgically resected intestine and mesentery there was vasculitis with fibrinoid necrosis accompanied by numerous eosinophils and macrophages containing eosinophil cationic protein (ECP). These findings suggest that in addition to ischemic changes due to vasculitis some neurotoxic substances generated by the eosinophils may be involved in the development of neuropathy in CSS. PMID- 12564772 TI - Sporadic amyotrophic lateral sclerosis with circumscribed temporal atrophy: a report of an autopsy case without dementia and with ubiquitinated intraneuronal inclusions. AB - This report concerns an autopsy case of amyotrophic lateral sclerosis (ALS) with circumscribed temporal atrophy. The patient was a Japanese woman without hereditary burden who was 71-year-old at the time of death. She developed dysarthria and gait disturbance at age 69, followed by dysphagia. A neurological examination about 1 year 11 months after the onset of the disease revealed absence of character change and of dementia. Neuroradiological examination disclosed circumscribed atrophy of the anterior part of the right temporal lobe. The patient died of respiratory failure 2 years after the disease onset. No respirator administration was performed throughout the clinical course. Macroscopically, neuropathological examination showed circumscribed atrophy of the right first temporal gyrus. Histologically, there was neuronal loss in the cerebral cortex, including the first temporal gyrus, the parahippocampal gyrus, subiculum, amygdala, substantia nigra, brain stem motor nuclei, and anterior horns of the spinal cord, in addition to loss of Betz cells, obvious degeneration of the pyramidal tracts, and the presence of Bunina bodies. Ubiquitin immunoreactive intraneuronal inclusions were present in the hippocampal dentate granular cells, frontotemporal cortical layer II neurons, and motor neurons in the brain stem and spinal cord. Based on these clinicopathological findings and a review of the literature, we concluded that our case was atypical ALS without dementia, showing temporal lobe atrophy macroscopically, in addition to pathological hallmarks compatible with ALS with dementia. We also note the possibility that there is a forme fruste of ALS with dementia showing no overt dementia clinically. PMID- 12564773 TI - Genotype-phenotype correlation in CAG-repeat diseases. AB - The expansion of a CAG repeat is a common causative gene mutation in several hereditary neurodegenerative disorders, including dentatorubral-pallidoluysian atrophy (DRPLA). Although, in DRPLA, it is revealed that the variety of clinical manifestations is related to the variable expansion of the CAG repeat, there are still many problems in the correlation between the symptoms and neuropathologic findings. Recent immunohistochemical studies have revealed that diffuse intranuclear accumulation of mutant proteins with expanded polyglutamine stretches is a significant pathology in DRPLA, and involves a wide range of the CNS regions far beyond the lesion distribution previously established by neuronal loss. This novel pathology may become a clue for elucidating molecular mechanisms of neuronal dysfunction and establishing clinicopathological correlations in CAG repeat diseases. PMID- 12564774 TI - Basal ganglia lesions in 'Pick complex': a topographic neuropathological study of 19 autopsy cases. AB - The distribution of basal ganglia lesions, including the amygdala, striatum (caudate nucleus, putamen), pallidum, and substantia nigra, were reinvestigated in 19 Japanese autopsy cases of 'Pick complex', consisting of five patients with corticobasal degeneration (CBD), 10 patients with Pick's disease with Pick bodies (PDPB), and four patients with generalized variant of Pick's disease (gvPD). The lesions in the amygdala, striatum, and pallidum were classified into three categories (severe, moderate, and slight). The lesions in the substantia nigra were qualitatively judged, compared with normal controls. In CBD, basal ganglia lesions in all five cases were uniform: the pallidum showed severe lesions, the striatum moderate lesions, the amygdala slight lesions, and obvious neuronal loss of the substantia nigra was verified in all five cases. Basal ganglia lesions in 10 cases of PDPB were also uniform: the amygdala disclosed severe to moderate lesions, the striatum moderate to slight lesions, the pallidum slight lesions, while obvious neuronal loss of the substantia nigra was found in only two of nine cases in which this structure was examined. Furthermore, basal ganglia lesions in all four cases of gvPD were uniform: the caudate nucleus showed severe lesions, the putamen and amygdala severe to moderate lesions, the pallidum moderate to slight lesions, and obvious neuronal loss of the substantia nigra was confirmed in all four cases. This study, using conventional staining such as hematoxylin eosin and Holzer, clarified that there were prominent lesions in the pallidum in CBD, in the amygdala in PDPB, and in the caudate nucleus in gvPD, respectively. In addition, nigral involvement was usually found in CBD and gvPD, but was rarely seen in PDPB. These neuropathological findings may help to elucidate the pathological heterogeneity of basal ganglia lesions in 'Pick complex'. PMID- 12564775 TI - Molecular biological approaches to neurological disorders including knockout and transgenic mouse models. AB - Advances of molecular biology have provided a great variety of new approaches to research on human disorders. This article gives an outline of molecular biological approaches to analysis of neurological disorders such as giant cell glioblastoma (GGBM) and amyotrophic lateral sclerosis (ALS), and their respective animal models: p53 knockout mice for GGBM and mutant superoxide dismutase-1 transgenic mice for ALS. Genomic DNA extracted from fresh-frozen tissue is examined by Southern blotting for screening mutations in a certain gene. Polymerase chain reaction (PCR) products of a gene in genomic DNA are examined by single-stranded conformation polymorphism, sequencing and agarose gel electrophoresis for identifying mutations, and for preparing and evaluating DNA probes used in Southern blotting and DNA in situ hybridization (ISH). Total RNA from tissue is examined by northern blotting for quantifying and verifying a certain mRNA. Reverse transcription-PCR products of a certain mRNA in total RNA are examined by sequencing and agarose gel electrophoresis for preparing and evaluating cDNA probes used in northern blotting and mRNA ISH. Tissue total protein is immunoblotted for quantifying and verifying a certain protein, and for evaluating the specificity of antibodies used in western blotting and immunohistochemistry. Immunoprecipitates are immunoblotted for evaluating a profile of protein or other substances. Enzyme-linked immunosorbent assay is used for measuring tissue concentration of protein or other substances, and for determining titers of specific antibodies. By these procedures, chronological analysis of animal models for human diseases contribute to elucidating pathogenic mechanisms and exploiting new therapies. Noticing both the similarity and difference between human and animal disorders will help understand the nature of disease. PMID- 12564776 TI - Nine-year-old girl presenting familial occurrence of progressive developmental abnormalities with the white matter lesions. PMID- 12564777 TI - 61-year-old woman presenting with deep coma during treatment for adrenal insufficiency. PMID- 12564778 TI - A 77-year-old woman presenting with removal of subcortical hemorrhages in the right parietal lobe. PMID- 12564779 TI - A 79-year-old woman with progressive cognitive impairment. PMID- 12564780 TI - A 76-year-old woman presenting with adult-onset, slowly progressive cerebellar symptoms. PMID- 12564781 TI - Left frontal tumor in a 46-year-old man with urinary incontinence and gait disturbance. PMID- 12564782 TI - Four-month-old male infant with generalized muscle weakness. PMID- 12564783 TI - 2002 Japanese Neuropathology Prize. Lectin cytochemistry of rat cerebral and cerebellar neuronal cells. PMID- 12564784 TI - Benefits of combination vaccines: effective vaccination on a simplified schedule. AB - BACKGROUND: The number of recommended immunizations during childhood has increased significantly over the years. Today, infants receive vaccines against 11 diseases in the first 2 years of life, and may receive as many as 5 injections at a single office visit. For a number of reasons, some injections may be deferred to a later visit, thus creating a missed opportunity for vaccination. OBJECTIVE: To review benefits of pediatric combination vaccines. RESULTS: Combination vaccines that protect against several diseases, such as the new DTaP HepB-IPV (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B [Recom-binant] and Inactivated Poliovirus Vaccine Combined) vaccine, can help to simplify the current immunization schedule. Other benefits of combination vaccines for parents and healthcare providers include decreased anxiety because of perceived reduction in pain for the infant, fewer missed opportunities to vaccinate, convenience, and decreased costs for the parent as a result of fewer office visits. Benefits of using combination vaccines in the office setting include reduced missed opportunities to vaccinate, storage of fewer vials, decreased risk of needle sticks as a result of handling fewer syringes, and potentially improved record keeping and tracking. Benefits of using combination vaccines in managed care include improved member satisfaction and lower costs because of fewer vaccine administration charges, and potentially fewer office visits. CONCLUSIONS: Combination vaccines are an effective means of decreasing the number of injections and simplifying the immunization schedule, thus providing overall benefits to infants, parents, healthcare providers, office managers, and managed care administrators. PMID- 12564785 TI - Clinical evaluation of a DTaP-HepB-IPV combined vaccine. AB - OBJECTIVE: To provide an overview of prelicensure clinical data for a new pediatric vaccine that combines diphtheria, tetanus, acellular pertussis, hepatitis B, and inactivated poliovirus vaccines into a single injection (DTaP HepB-IPV combined vaccine; Pediarix, GlaxoSmithKline Biologicals, Rixensart, Belgium). METHODS: The safety and immunogenicity of DTaP-HepB-IPV combined vaccine have been evaluated extensively in clinical trials in infants. To date, DTaP-HepB-IPV combined vaccine has been administered to more than 7,000 infants as a 3-dose primary series during the first year of life. RESULTS: Studies show that DTaP-HepB-IPV combined vaccine is generally safe, well tolerated, and has not caused any significant serious adverse events. The rates of solicited and unsolicited reports of adverse reactions following vaccination were similar between groups receiving DTaP-HepB-IPV combined vaccine and comparator groups receiving the vaccine components separately. DTaP-HepB-IPV combined vaccine induces immunogenicity (as measured by seroprotection or vaccine response rates to each of the vaccine components [diphtheria, tetanus, 3 pertussis antigens, hepatitis B, and poliovirus types 1, 2, and 31) similar to licensed vaccine components administered separately. CONCLUSION: In prelicensure clinical studies, DTaP-HepB-IPV combined vaccine was safe and immunogenic when given to infants as a primary 3-dose series. As a single injection of multiple vaccine components, DTaP-HepB-IPV combined vaccine may provide a safe and effective alternative to the current multiple-injection immunization regimen. PMID- 12564786 TI - Considerations for implementing a new combination vaccine into managed care. AB - BACKGROUND: The control and elimination of several deadly childhood diseases are a result of extensive vaccination efforts made by pediatricians, family practitioners, public health providers, and health outreach systems. Managed care can assist this effort through facilitating the delivery of affordable quality healthcare to patients. OBJECTIVE: To describe the considerations made by managed care when implementing a new vaccine into practice. RESULTS: Managed care plans often assess the medical necessity, consumer acceptance, and pharmacoeconomic benefits of a vaccine when considering whether it can be implemented into practice. DTaP-HepB-IPV (Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed, Hepatitis B [Recombinant] and Inactivated Poliovirus Vaccine [Combined]), a new combination vaccine, has demonstrated similar immunogenicity and safety when compared with separately administered component vaccines. Use of this combination vaccine will help to simplify the current immunization schedule, and therefore decrease the number of injections infants receive in the first year of life, favorably influencing consumer perception of this new vaccine. A reduction in the number of office visits as a result of fewer injections and improved vaccine compliance may result in a positive pharmacoeconomic impact on parents, physicians, and payers. Reduced administration fees, fewer needed syringes, and decreased risk of needlestick injury resulting from the use of combination vaccines all may have a positive impact on acceptance of these vaccines by managed care organizations. CONCLUSION: DTaP-HepB-IPV is expected to meet the tests of medical appropriateness, consumer acceptance, and pharmacoeconomic reasonableness, thereby fulfilling the value proposition of a new combination vaccine for managed healthcare plans. PMID- 12564787 TI - The future of childhood immunizations: examining the European experience. AB - BACKGROUND: New pediatric combination vaccines that protect against multiple diseases are being introduced into clinical practice in the United States. Similar vaccines have been used extensively throughout Europe and other parts of the world for the past few years. OBJECTIVE: The objective of this article is to comment on the use in Europe of pediatric combination vaccines that protect against multiple diseases to predict their acceptance and use in the United States. RESULTS: Throughout Europe, combination vaccines that safely and effectively protect against multiple diseases have been well received, are widely used, and have helped greatly to simplify pediatric immunization schedules. In Germany, for example, the use of hexavalent combination vaccines has simplified the immunization schedule so that infants now receive a maximum of 2 injections per office visit, compared with the up to 5 injections per visit that infants may receive in the United States. Because of the many advantages and wide acceptance of combination vaccines in Europe, it is anticipated that similar vaccines will be readily accepted in the United States. CONCLUSIONS: As demonstrated in Europe and elsewhere, combination vaccines are an efficient means of simplifying the pediatric immunization schedule, and the new combination vaccines being introduced in the United States are expected to be well received. PMID- 12564788 TI - Effect of nitrogen and water treatment on leaf chemistry in horsenettle (Solanum carolinense), and relationship to herbivory by flea beetles (Epitrix spp.) and tobacco hornworm (Manduca sexta). AB - We studied the interaction between plants (horsenettle; Solanum carolinense) and herbivorous insects (flea beetles; Epitrix spp., and tobacco hornworm; Manduca sexta) by focusing on three questions: (1) Does variation in nitrogen availability affect leaf chemistry as predicted by the carbon-nutrient balance (CNB) hypothesis? (2) Does variation in plant treatment and leaf chemistry affect insect feeding? (3) Is there an interaction between the insect herbivores that is mediated by variation in leaf chemistry? For three successive years (1998-2001), we grew a set of clones of 10 maternal plants under two nitrogen treatments and two water treatments. For each plant in the summer of 2000, we assayed herbivory by hornworms in both indoor (detached leaf) and outdoor (attached leaf) assays, as well as ambient flea beetle damage. Estimates of leaf material consumed were made via analysis of digitized leaf images. We also assayed leaves for total protein, phenolic, and glycoalkaloid content, and for trypsin inhibitor, polyphenol oxidase, and peroxidase activity. Despite strong effects of nitrogen treatment on growth and reproduction, only total protein responded as predicted by CNB. Leaf phenolic levels were increased by nitrogen treatment, polyphenol oxidase activity was decreased, and other leaf parameters were unaffected. Neither hornworm nor flea beetle herbivory could be related to plant treatment or genotype or to variation in any of the six leaf chemical parameters. A negative relationship between flea beetle and hornworm herbivory was found, but was not apparently mediated by any of the measured leaf chemicals. Because leaf resistance was maintained in low nitrogen plants at the apparent expense of growth and reproduction, our results support the concept of a fitness cost of defense, as predicted by the optimal defense hypothesis. PMID- 12564789 TI - Relationship between nutritional composition of plant species and infestation levels of thrips. AB - Levels of soluble protein and carbohydrate (raffinose, sucrose, glucose, and fructose) in leaves from a selection of plant species were measured to determine if a relationship existed between these nutrients and infestation by Frankliniella occidentalis and Heliothrips haemorrhoidalis. Most species of host plant examined contained a higher proportion of protein than carbohydrates, and overall, leaves from species of plants that supported populations of thrips had greater levels of protein than leaves from nonhost species. New leaves and flowers that supported F. occidentalis contained high levels of carbohydrate and protein. The quantity of protein in leaves at the top of the tree, Peumus boldus, was greater than in leaves from lower levels, and the amount of feeding damage accrued by H. haemorrhoidalis was greater on the upper foliage than lower foliage. Oviposition by H. haenmorrhoidalis was positively correlated to levels of protein in host plants but not to levels of carbohydrates. Overall, levels of soluble protein in plants influenced their susceptibility to thrips more than levels of carbohydrates. PMID- 12564790 TI - Monoterpene effect on feeding choice by deer. AB - A previous study showed that Sitka black-tailed deer (Odocoileus hemionus sitkensis) consumption was negatively correlated with monoterpene content in western redcedar (Thuja plicata). To test whether these monoterpenes were deterrent to Sitka black-tailed deer, we performed feeding choice experiments with four hydrocarbon (sabinene, myrcene, alpha-pinene, and d + l-limonene) and one oxygenated (alpha,beta-thujone) monoterpene solution at their highest natural concentration in western redcedar foliage. To test whether deer response was species specific, we ran similar experiments on European roe deer (Capreolus capreolus) and rusa deer (Cervus timorensis russa). In all experiments, monoterpenes were repellent. Solutions with alpha,beta-thujone, the major monoterpene in redcedar leaves, were the most repellent of the solutions tested. We then analyzed how black-tailed and roe deer responded to (1) an increase in concentration of the monoterpenes with the weakest repellent effects (hydrocarbon monoterpenes) and (2) a decrease in concentration of the monoterpene with strongest effect (alpha,beta-thujone). Repellency tended to increase with concentration for hydrocarbon monoterpenes, but remained strong for alpha,beta thujone. As wild deer regularly feed on plants containing monoterpenes, this raises the question as to how the animals deal with these molecules. PMID- 12564791 TI - Systemic, genotype-specific induction of two herbivore-deterrent iridoid glycosides in Plantago lanceolata L. in response to fungal infection by Diaporthe adunca (Rob.) Niessel. AB - Iridoid glycosides are a group of terpenoid secondary plant compounds known to deter generalist insect herbivores. In ribwort plantain (Plantago lanceolata), the iridoid glycosides aucubin and catalpol can be induced following damage by insect herbivores. In this study, we investigated whether the same compounds can be induced following infection by the fungal pathogen Diaporthe adunca, the causal agent of a stalk disease in P. lanceolata. Significant induction of aucubin and catalpol was observed in two of the three plant genotypes used in this study following inoculation with the pathogen. In one of the genotypes, induction occurred within 6 hr after inoculation, and no decay was observed within 8 days. The highest level of induction was observed in reproductive tissues (spikes and stalks) where infection took place. In these tissues, iridoid glycoside levels in infected plants were, on average, 97% and 37% higher than the constitutive levels in the corresponding control plants, respectively. Significant induction was also observed in leaves (24%) and roots (17%). In addition to significant genotypic variation in the level of induction, we found genetic variation for the tissue-specific pattern of induction, further broadening the scope for evolutionary fine-tuning of induced responses. Recent studies have revealed a negative association between iridoid glycoside levels in P. lanceolata genotypes and the amount of growth and reproduction of D. adunca that these genotypes support. However, for the three genotypes used in the present study, differences in resistance were not related to their constitutive or induced levels of iridoid glycosides, suggesting that additional resistance mechanisms are important in this host-pathogen system. We conclude that iridoid glycosides in P. lanceolata can be induced both by arthropods and pathogenic micro-organisms. Pathogen infection could, therefore, potentially enhance resistance to generalist insect herbivores in this species. PMID- 12564792 TI - Comparative physiological responses in Chinese cabbage induced by herbivory and fungal infection. AB - Fungal infection of Chinese cabbage leaves by Alternaria brassicae has earlier been shown to have detrimental effects on larval development of the chrysomelid beetle Phaedon cochleariae. Furthermore, adults of this leaf beetle avoid fungus infected Chinese cabbage leaves for oviposition and feeding. However, herbivory had no impact on fungal growth. In this study, we investigated physiological responses of the host plant to herbivore attack and fungal infection in order to elucidate the mechanisms of the described ecological interactions between the fungus and the herbivore. Changes in primary factors (water, C/N ratio, protein, sucrose) and defense-related plant compounds (glucosinolates, anthocyanins, peroxidase) were measured. Herbivory and fungal infection reduced the sucrose concentration of leaves and increased amounts of indole glucosinolates as well as total anthocyanins. In addition, water content was slightly lower in insect damaged but not in infected leaves. Higher levels of peroxidase activity resulted exclusively from fungal infection. The C/N ratio and total protein content remained unaffected by either treatment. The implications of the induced plant changes on the herbivore are discussed. PMID- 12564793 TI - Predatory response of brown tree snakes to chemical stimuli from human skin. AB - The brown tree snake (Boiga irregularis) is an exotic pest species on Pacific islands, most notably on Guam where it has caused considerable ecological and economic damage. On Guam, the snake commonly associates with people and can be found near or in human habitations. Bites are common, approximately 1 of 1200 emergency room visits to Guam hospitals were reported to be the result of B. irregularis bites; 80% of these victims were attacked while sleeping. Most of the attacks occurred on fingers and hands and the attacks appeared to be predatory, rather than defensive, in nature. In order to characterize the mechanism releasing this unusual behavior, we measured the predatory response of B. irregularis to chemical stimuli from humans and controls using a lab population that originated from Guam and a wild population from the species' native range in Queensland, Australia. To quantify behavior we measured the proportion of snakes displaying predatory behavior to each of the stimuli, the latency to attack, and the number of tongue-flicks displayed. We quantified predatory behavior using the tongue-flick attack score for repeated measures [TFAS(R)], a common method for quantifying predatory behavior in squamate reptiles. Captive brown tree snakes responded to human skin stimuli with feeding behavior, including predatory attacks, at the same frequency as they did to prey stimuli derived from mice, while never responding to controls with such behavior. Captive snakes also responded to human skin stimuli and prey stimuli with significantly higher TFAS(R) scores than to controls, although there were no differences between the human and mouse stimuli. Wild-caught animals in Australia also responded with predatory attacks to human skin stimuli, while not showing predatory behavior to a blank control and with higher TFAS(R) scores to human skin stimuli than to the control. As B. irregularis is a generalized predator that relies heavily on chemical signals to recognize prey, we hypothesize that the snakes recognize compounds on human skin that may be shared with other prey. PMID- 12564794 TI - Transport of phenolic compounds from leaf surface of creosotebush and tarbush to soil surface by precipitation. AB - During the last 100 years, many desert grasslands have been replaced by shrublands. One possible mechanism by which shrubs outcompete grasses is through the release of compounds that interfere with neighboring plants. Our objective was to examine the movement of secondary compounds from the leaf surface of creosotebush and tarbush to surrounding soil surfaces via precipitation. Units consisting of a funnel and bottle were used to collect stemflow, throughfall, and interspace precipitation samples from 20 creosotebush (two morphotypes) and 10 tarbush plants during three summer rainfall events in 1998. Precipitation samples were analyzed for total phenolics (both species) and nordihydroguaiaretic acid (creosotebush only). Phenolics were detected in throughfall and stemflow of both species with stemflow containing greater concentrations than throughfall (0.088 and 0.086 mg/ml for stemflow and 0.022 and 0.014 mg/ml for throughfall in creosotebush morphotypes U and V, respectively: 0.044 and 0.006 mg/ml for tarbush stemflow and throughfall, respectively). Nordihydroguaiaretic acid was not found in any precipitation collections. The results show that phenolic compounds produced by creosotebush and tarbush can be transported to the soil surface by precipitation, but whether concentrations are ecologically significant is uncertain. Nordihydroguaiaretic acid was not present in the runoff from creosotebush. PMID- 12564795 TI - Comparison of different methods for lignin determination as a basis for calibration of near-infrared reflectance spectroscopy and implications of lignoproteins. AB - Three commonly employed methods for lignin determination, i.e., the thioglycolic acid (TGA), the acetylbromide (AB), and the acid detergent fiber (ADF) method, were compared using leaves and xylem tissue from five species (Nicotiana tabacum, Populus x canescens, Fagus sylvatica, Quercus robur, and Picea abies). In each case, cell walls were isolated before lignin determination. Each of the three methods estimated a different lignin concentration in a given tissue, except for spruce wood. The "lignin" concentration determined with the AB method was strongly dependent on whether or not the cell walls were subjected to alkaline hydrolysis to remove covalently bound aromatic nonligneous components before lignin determination. Lignin concentrations determined in hydrolyzed cell walls of different tissues and species by the AB method showed a good correlation with those obtained by the TGA method and, thus, were convertible. In contrast, gravimetrically estimated ADF lignins did not or only moderately correlate with lignins measured with methods based on the UV absorbance of the solubilized lignin degradation products. Leaves of a given species generally contained higher ADF-lignin concentrations than the corresponding stem tissue. Both ADF and TGA lignin data of beech were used to calibrate near-infrared reflectance spectra (NIRS) for lignin prediction. Both NIRS calibration procedures gave good statistical fits with correlation coefficients close to 1, indicating that TGA and ADF lignin concentrations of beech can be estimated by NIRS with high accuracy. However, the two calibrations were based on different empirical terms, showing that TGA and ADF lignins did not share the same physical basis for calibration. C/N analysis revealed the presence of 3.1 and 1.4% nitrogen in ADF lignins of beech leaves and wood, respectively. The major fraction of this nitrogen was recovered in amino acids, which corresponded to 14% and 3% protein in ADF lignins of leaves and wood, respectively. These results show that ADF lignins contain significant concentrations of lignin-bound proteins, which renders this method unsuitable to determine genuine lignin. PMID- 12564796 TI - Bruchins, plant mitogens from weevils: structural requirements for activity. AB - Bruchins are 3-hydroxypropanoate esters of long-chain alpha,omega-diols from pea weevils and cowpea weevils that have been shown to initiate callus formation on pea pods at extremely low application rates. Synthetic analogs have been prepared and examined to evaluate structural requirements for inducing this unusual neoplastic response. Chain length (optimum length C22-C24) is important, whereas unsaturation within the chain is relatively unimportant. Difunctionality is required for maximum activity, but the alpha,omega-diols themselves are inactive. Most critical is the ester portion(s) of the molecules; 3-hydroxypropanoate esters are far more active than any analogs examined. PMID- 12564797 TI - Right stereoisomers for sex pheromone components of the apple leafminer, Lyonetia prunifoliella, in Korea. AB - The sex pheromone of the American population of Lyonetia prunifoliella (Lepidoptera: Lyonetiidae) was reported to be composed of three components, 10,14 dimethyloctadec-1-ene (10me14me-1-ene-18Hy), 5,9-dimethyloctadecane (5me9me 18Hy), and 5,9-dimethylheptadecane (5me9me-17Hy). Studies with a Korean population showed that the 10S, 14S isomer of the major compound was the major electrophysiologically by active stereoisomer. The 5S,9S isomers of the minar constituents also elicited electrophysiological responses. In field trappings, among the three components, the racemic mixture of 10me14me-1-ene-18Hy attracted L. prunifoliella males strongly, and the binary or tertiary combinations with racemic mixtures of the other two components did not increase trap catches. Among the four stereoisomers of 10me14me-1-ene-18Hy, only the 10S,14S isomer attracted L. prunifoliella males, as suggested by the EAG test, and attraction to the other three isomers was not observed. All stereoisomers of each minor component, as well as their racemic components, did not seem to have any additional effect on male attraction in Korea. PMID- 12564798 TI - Semiochemicals of the Scarabaeinae. VII: Identification and synthesis of EAD active constituents of abdominal sex attracting secretion of the male dung beetle, Kheper subaeneus. AB - Using gas chromatography with flame ionization detection (FID) and electroantennographic detection (EAD) in parallel, butanoic acid, skatole, and (E)-2,6-dimethyl-6-octen-2-ol were identified as constituents of the abdominal sex-attracting secretion of the male dung beetle, Kheper subaeneus, which reproducibly elicited EAD responses in male and female antennae. This is the first report of the occurrence of (E)-2,6-dimethyl-6-octen-2-ol as a natural product, for which the name (E)-subaeneol is proposed. In some experiments, a few other constituents of the secretion also gave reproducible responses in specific male and female antennae but did not elicit responses when the analyses were repeated with other antennae. The major volatile constituent of the secretion, identified as (S)-(+)-2,6-dimethyl-5-heptenoic acid, is one of these EAD-active compounds. Both this compound and (E)-2,6-dimethyl-6-octen-2-ol were synthesized from authentic starting materials for comparison with the natural products. PMID- 12564799 TI - Identification of new homoterpene esters from Dufour's gland of the ponerine ant Gnamptogenys striatula. AB - Extracts of Dufour's gland of the ponerine ant, Gnamptogenys striatula, were analyzed by using the combination of gas chromatography and mass spectrometry. Series of esters of the new homoterpenoids (2E,6)-3,4,7-trimethyl-2,6-octadiene-1 ol (4-methylgeraniol) and (2E,6)-3,4,7-trimethyl-2,6-nonadiene-1-ol (bishomogeraniol) with unbranched medium-chain fatty acids were identified. Transformation of the chiral natural products into 1,4-di (trifluoroacetoxy)-3 methylpentane and comparison of its gas chromatographic retention time on a modified cyclodextrin phase with that of synthetic optically active reference samples proved the stereogenic center to keep (S)-configuration. (2E,4S,6)-3,4,7 Trimethyl-2,6-octadien-1-yl decanoate and the corresponding dodecanoate are the main volatiles in the extracts. PMID- 12564800 TI - Trail pheromone of ponerine ant Gnamptogenys striatula: 4-methylgeranyl esters from Dufour's gland. AB - Dufour's gland is the origin of the trail pheromone of Gnamptogenys striatula. Chemical analysis of the glandular extracts revealed a series of new natural products, especially esters of (2E)-3,4,7-trimethyl-2,6-octadien-1-ol (4 methylgeraniol), and (2E)-3,4,7-trimethyl-2,6-nonadien-1-ol (a bishomogeraniol isomer) with medium-chain fatty acids. Bioassays with synthetic racemates of the esters revealed that the 4-methylgeranyl esters are highly active as trail pheromones, while the bishomogeranyl esters are either marginally active or not active at all. Assays with the individual 4-methylgeranyl esters showed each of them to be inferior to the glandular secretion in eliciting trail following. However, the mixture of racemic 4-methylgeranyl octanoate and the corresponding decanoate and dodecanoate, the main Dufour's volatile constituents, is as active as the natural secretion at similar concentration. We conclude that the trail pheromone constitutes a mixture of at least the 4-methylgeranyl esters identified in the gland. Since G. striatula generally preys on small arthropods rather than monopolizing large resources, we assume that trails are rarely used during foraging, but more often during nest migration. Production of new societies in this species is generally performed by budding, a period of considerable predation risk. Utilizing trails for efficient displacement in this context is, therefore, highly adaptive. This behavioral repertoire may also provide the ants with additional means of food resource exploitation. PMID- 12564801 TI - Cuticular hydrocarbons of Tetramorium ants from central Europe: analysis of GC-MS data with self-organizing maps (SOM) and implications for systematics. AB - Cuticular hydrocarbons were extracted from workers of 63 different nests of five species of Tetramorium ants (Hymenoptera: Formicidae) from Austria, Hungary, and Spain. The GC-MS data were classified (data mining) by self-organizing maps (SOM). SOM neurons derived from primary neuron separation were subjected to hierarchical SOM (HSOM) and were grouped to neuron areas on the basis of vicinity in the hexagonal output grid. While primary neuron separation and HSOM resulted in classifications on a level more sensitive than species differences, neuron areas resulted in chemical phenotypes apparently of the order of species. These chemical phenotypes have implications for systematics: while the chemical phenotypes for T. ferox and T. moravicum correspond to morphological determination, in T. caespitum and T. impurum a total of six chemical phenotypes is found. Three hypotheses are discussed to explain this disparity between morphological and chemical classifications, including in particular the possibility of hybridization and the existence of cryptic species. Overall, the GC-MS profiles classified by SOM prove to be a practical alternative to morphological determination (T. ferox, T. moravicum) and indicate the need to revisit systematics (T. caespitum, T. impurum). PMID- 12564802 TI - Semiochemicals released by electrically stimulated red imported fire ants, Solenopsis invicta. AB - The red imported fire ant Solenopsis invicta Buren, has evolved sophisticated chemical communication systems that regulate the activities of the colony. Among these are recruitment pheromones that effectively attract and stimulate workers to follow a trail to food or alternative nesting sites. Alarm pheromones alert, activate, and attract workers to intruders or other disturbances. The attraction and accumulation of fire ant workers in electrical equipment may be explained by their release of pheromones that draw additional worker ants into the electrical contacts. We used chemical analysis and behavioral bioassays to investigate if semiochemicals were released by electrically shocked fire ants. Workers were subjected to a 120 V, alternating-current power source. In all cases, electrically stimulated workers released venom alkaloids as revealed by gas chromatography. We also demonstrated the release of alarm pheromones and recruitment pheromones that elicited attraction and orientation. Arrestant behavior was observed with the workers not electrically stimulated but near those that were, indicating release of unkown behavior-modifying substances from the electrically stimulated ants. It appears that fire ants respond to electrical stimulus by generally releasing exocrine gland products. The behaviors associated with these products support the hypothesis that the accumulation of fire ants in electrical equipment is the result of a foraging worker finding and closing electrical contacts, then releasing exocrine gland products that attract other workers to the site, who in turn are electrically stimulated. PMID- 12564803 TI - 1-Octen-3-ol together with geosmin: new secretion compounds from a polydesmid millipede, Niponia nodulosa. AB - The small millipede Niponia nodulosa (Polydesmida: Cryptodesmidae) emits an earthy smell when disturbed. This smell was obtained from hexane extracts from both sexes at the sixth and seventh instars and adult stages, and was found by GC/MS analyses to be composed of two compounds, 1-octen-3-ol as the major component and geosmin as the minor component. This is the first report of these compounds in secreted substances from millipedes. As they showed little repellent activity against foraging ants in bioassays, their biological function as defensive allomones was not clearly elucidated. However, both compounds provoked negative chemotaxis of conspecific millipedes in a Y-tube olfactometer, especially 1-octen-3-ol, which was active at a dose equivalent to the content of a single millipede. Such biological activity suggests that this compound is involved in intraspecific communication in this millipede and may play a role as an alarm pheromone. PMID- 12564804 TI - Semiochemicals and social signaling in the wild European rabbit in Australia: II. Variations in chemical composition of chin gland secretion across sampling sites. AB - The volatile components of the chin gland secretion of the wild European rabbit, Oryctolagus cuniculus (L.), were investigated with the use of gas chromatography. Studies of the chemical nature of this secretion by previous workers demonstrated that it was important in the maintenance of social structure in this species. This study identified 34 different volatile components that consist primarily of aromatic and aliphatic hydrocarbons. Especially common are a series of alkyl substituted benzene derivatives that provide most of the compound diversity in the secretion. Samples of chin gland secretion collected from animals at three different geographical locations, separated by more than 100 km, showed significant differences in composition. This work suggests that variation among populations needs to be considered when undertaking semiochemical research. Alternate nonparametric methods are also used for the analysis of chromatographic data. PMID- 12564805 TI - Chemical polymorphism of the cuticular lipids of the cabbage white Pieris rapae. AB - The epicuticular composition of different body parts of the Cabbage White, Pieris rapae L., was investigated using GC and GC/MS. The major group of components, hydrocarbons, occurs in two distinct classes, which show different distributions on the cuticle of the insects. Unbranched shorter chain compounds (C21 to C31, linear group) dominate on body, head and wings, while longer chain, polymethyl branched compounds (C35 to C39, branched group) are predominantly found on the antennae. Several other components like 1,3-pentacosadiene and oxygenated aliphatic compounds occur in minor amounts on the cuticle. The reason for this polymorphism is discussed. PMID- 12564806 TI - Disease-modifying therapy in relapsing--remitting multiple sclerosis: efficacy is paramount. AB - The primary objective in the management of a chronic disease is to maximise therapeutic effectiveness, according to the general consensus among specialists. Recent market research has confirmed that a treatment's effectiveness is the primary consideration for neurologists' choice of therapy for multiple sclerosis (MS). Of the available disease-modifying therapies, interferon (IFN) beta appears to be consistently more efficacious than glatiramer acetate. High doses of therapy, and frequent administration of IFN beta should be used to provide maximal effects. This has been supported by a recent Class I comparative trial of two commercial preparations of IFN beta-1a. Preliminary results indicated significantly greater efficacy for IFN beta-1a (Rebif, Serono) 44 microg administered subcutaneously three times weekly (t.i.w.), over IFN beta-1 (Avonex, Biogen) 30 microg administered intramuscularly once weekly IFN beta-1a, 44 microg t.i.w., provides maximal efficacy for patients with relapsing forms of MS. PMID- 12564807 TI - Once weekly interferon beta for multiple sclerosis is superseded by higher and more frequent dosing. AB - Therapy with interferon (IFN) beta can significantly alter the disease course in relapsing-remitting multiple sclerosis. Evidence indicates that the efficacy of this agent is related to treatment regimen (dose, route and dose frequency). A higher total weekly dose administered several times a week provides greater clinical benefit than a lower dose given once weekly (q.w.). This dose- and dose frequency-dependency of the efficacy of IFN beta has been clearly demonstrated in three recent head-to-head studies, as well as in other major clinical trials, and pharmacodynamic, pharmacokinetic and pre-clinical studies. The use of a q.w. dose regimen of IFN beta is likely to have a negative impact on patients in both the short and long-term because it allows additional accumulation of irreversible tissue damage in the central nervous system. Patients should receive the optimal, more rapid benefits from higher and more frequent doses of IFN beta, administered as early as possible after diagnosis. PMID- 12564808 TI - Early use of interferon beta patients with multiple sclerosis. AB - In recent years, major advances have been made in the management of multiple sclerosis (MS) in the form of new disease-modifying therapies, the most widely used of which is interferon (IFN) beta. A growing body of evidence indicates that the beneficial effects of IFN beta are maximised if treatment is started as soon as possible after the diagnosis of MS, and if patients are given the highest possible dose. The argument in favour of early treatment is based primarily on the finding that the inflammation of the central nervous system characteristic of MS leads to irreversible axonal destruction starting very early in the course of the disease. Evidence that IFN beta should be given at high doses comes from preclinical and clinical studies showing that the effects of this drug are strongly dose-dependent. Three formulations of IFN beta are currently available for the treatment of MS: subcutaneous (s.c.) IFN beta-1a, intramuscular (i.m.) IFN beta-1a and s.c. IFN beta-1b. All are well tolerated, although IFN beta-1a appears to be less immunogenic than IFN beta-1b. IFN beta-1a, in s.c. formulation, has advantages over the other formulations in terms of convenience, and is approved for use at higher doses than i.m. IFN beta-1a. PMID- 12564809 TI - Interferon beta and multiple sclerosis: look at the evidence. AB - Recent advances in therapy for multiple sclerosis (MS) have centred on the use of the disease-modifying drugs glatiramer acetate (GA) and interferon (IFN) beta. Several large-scale clinical trials have been carried out on the use of these compounds, but there have been few studies that have directly compared their efficacy in MS. Furthermore, there has been controversy and confusion over the IFN beta therapy regimen that will achieve the best possible clinical outcome for MS patients. This review focuses principally on clinical trials of IFN beta-1a, where data that allow direct comparison of different treatment regimens are now available. Current data indicate that IFN beta, and in particular IFN beta-1a, has important advantages over GA in the treatment of relapsing-remitting MS (RRMS). Additionally, IFN beta-1a (Rebif, Serono), 44 microg administered subcutaneously (s.c.) three times weekly (t.i.w.), is significantly more effective than IFN beta-1a (Avonex, Biogen), 30 microg administered intramuscularly once weekly. For optimal management of RRMS, treatment with IFN beta-1a, 44 microg s.c. t.i.w., should begin as early as possible after diagnosis. PMID- 12564810 TI - In vivo evaluation of leucocyte dynamics in cremaster muscle in rats after exposure to cigarette smoke. AB - This experiment was designed to examine the effects of chronic cigarette smoking on the microcirculation immediately after exposure to smoke for two, four, and six weeks and after withholding smoke for two weeks from those previously exposed for four weeks. The mean (SD) rolling leucocytes at two, four, and six weeks were 11.10 (1.8), 23.7 (2.3), 40.2 (3.9) (p < 0.001). The rolling leucocytes after smoking for four weeks and then having smoke withheld for two weeks was 9.6 (1.4). The mean adherent leucocytes were 5.0 (0.7), 7.5 (1.1), 12.6 (1.8) (p < 0.001). The adherent leucocytes after smoking for four weeks and then having smoke withheld for two weeks was 3.5 (0.5). Our results confirm those of many previous studies of the adverse effects of cigarette smoking, and also that those deleterious effects are time-dependent. The reversibility of the deleterious effect of cigarette smoking after cessation of cigarette smoking before face-lift or flap reconstruction is at least two weeks. This information is also important for clinical management of patients who smoke and who are scheduled for face-lift and flap reconstruction. Two weeks without cigarettes is a necessary period for successful elective plastic surgery. PMID- 12564811 TI - Microsurgical tissue transfer and individual computer-aided designed and manufactured prefabricated titanium implants for complex craniofacial reconstruction. AB - From 1994 to 2000 187 individual computer-aided designed and manufactured (CAD/CAM) prefabricated titanium skull implants were inserted at 37 clinical centres. Since the processing chain of construction and fabrication of implants has become routine the clinical success totally depends on the condition of the soft tissues at the recipient site. In three patients in our own department with a history of up to 18 surgical interventions, and additional previous irradiation in one case, these conditions were so bad that a microsurgical tissue transfer had to be made before insertion of the implant. A latissimus dorsi free flap with submandibular microsurgical anastomosis had to be used in all three cases. However, the aetiology of the soft tissue deficits differed, and they were at a different tissue level in each case: anterior skull base, subcutaneous temporal area, and frontoparietotemporal skin. This series of patients therefore demonstrates the variability of possible combinations, which also require special timetables and principles of construction of the CAD of the implant. In all cases the cranioplasties were done three to five months after the transfer of the flaps and fulfilled the criteria of greatest precision and the best possible aesthetic outcome with minimal stress for the patients. These applications are surgical strategies for extreme cases but also illustrate the elaborate interdisciplinary approach in Computer Assisted Surgery. PMID- 12564812 TI - Unrepaired clefts in the hard palate: speech deficits at the ages of 5 and 7 years and their relationship to size of the cleft. AB - The purpose of the study was to investigate the relationship between the size of oronasal openings in the hard palate and speech deficits. Audiotape recordings and plaster casts were taken according to standard procedures at 5 and 7 years of age from 22 consecutive children born with complete unilateral cleft lip and palate treated at Sahlgrenska University Hospital, Goteborg, Sweden. The soft palate had been repaired before the age of 12 months, whereas the cleft in the hard palate was left unrepaired, to be closed later. Perceptual judgements of nine speech variables at 5 and 7 years of age were correlated with measures of the area of the residual cleft in the hard palate. "Retracted oral articulation" (to velar place) found in nine of the 22 children correlated significantly with the area of the cleft at the age of 5 years but not later. The establishment of this particular speech error seems to be related to the size of an oronasal opening. PMID- 12564813 TI - Transplantation of autologous cultivated conjunctival epithelium for the restoration of defects in the ocular surface. AB - Ocular surface disorders are often characterised by partial or complete loss of corneo-conjunctival epithelium, which causes dramatic functional and cosmetic problems. Depletion of stem cells and the scarcity of donor tissue available make large or bilateral defects challenging to reconstruct, and usually require the transplantation of heterotopic or allogeneic grafts. We investigated the feasibility of restoring severely damaged ocular surfaces with autologous cultivated conjunctival epithelium. Conjunctival cells were harvested from the healthy eyelid bed of four patients with oculopalpebral diseases. An epithelial sheet reproducing the original conjunctival epithelium was generated by serial cell culture. This was transplanted for the first time ever to our knowledge on to the ocular surface of the same patients. Take was excellent and the cultivated epithelium was stable, resulting in great improvement of patient's symptoms and cosmesis. Transplantation of cultivated conjunctival cells opens new perspectives in the treatment of severe ocular surface disorders. PMID- 12564814 TI - Osteotomy of the humerus to improve external rotation in nine patients with brachial plexus palsy. AB - Malfunction of the infraspinatus muscle and teres minor muscle illustrate the typical clinical picture in patients with brachial plexus palsy. The arm hangs down in an inwardly rotated position and elbow flexion is hindered by striking of the lower arm against the thorax. Between 1995 and 2000, we have done external rotational osteotomy of the humerus for nine patients with brachial plexus palsy. The mean age of the patients at the time of operation was 29 years (range 15 to 42). The mean follow-up time was 24 (6 to 69) months. Preoperatively, the patients all had appreciable deficits of external rotation (mean deficit 37 degrees, range 10 degrees to 70 degrees). As a result of osteotomy, external rotation was improved in all patients, the mean increase being 42 degrees (range 25 degrees to 60 degrees). All patients were subjectively content with the improved position of the arm and its function. They were able to move their hands to their faces without striking the lower arm against the chest on elbow flexion, or without compensatory evasive movement of the shoulder. PMID- 12564815 TI - Prospective comparative study of the range of movement of temporomandibular joints after mandibular fractures: rigid or non-rigid fixation. AB - Temporomandibular joints (TMJ) are negatively affected by trauma and disuse. In this prospective study, 103 patients with mandibular fractures were evaluated for the influence of trauma and maxillomandibular fixation on the TMJ. A total of 54 patients were treated by maxillomandibular fixation and 49 by titanium miniplate fixation. Those patients with condylar fractures and multiple mandibular fractures and malocclusion at presentation were not included in the study. The control group consisted of 44 randomly-selected healthy people with no past history of mandibular fracture or symptoms referable to the TMJ. Trauma was a major factor leading to TMJ dysfunction and maxillomandibular fixation increased the incidence and severity of TMJ dysfunction. PMID- 12564816 TI - Recovery of sensitivity in the hand after reconstruction with arterialised venous flaps. AB - Loss of soft tissues of the fingers were repaired in 22 patients using 25 arterialised venous flaps harvested from the thenar, hypothenar, or forearm regions. Twenty-one of the flaps survived completely, 16 of which were raised from the thenar or hypothenar region, and the other five from the forearm region. We studied the sensory recovery and skin characteristics of the flaps harvested from the three regions. Good sensory recovery was obtained for the thenar or hypothenar venous flaps, which were characterised by durable skin and suitable texture for replacement of defects in the finger pulp. On the other hand, no moving two-point discrimination was recorded during the follow-up period in the group given forearm venous flaps. These flaps showed instability during pinching and grasping. However, larger flaps and longer veins can be harvested from the forearm region. This type of flap is therefore considered useful for covering dorsal defects of the finger or large and multiple skin defects. PMID- 12564817 TI - Correction of a deformed thumb by distraction of the phalanx. AB - We used distraction osteogenesis to correct six deformed thumbs in four patients ranging in age from 4 to 7 years. Two of the patients had Apert syndrome (syndromic craniosynostosis with symmetrical syndactyly) and two had polydactyly. We used a small fixator with a ball joint and successfully corrected the angular deformity after lengthening the proximal phalanx by distraction. This single inclusive procedure was extremely useful. We found the optimal distraction regimen for the digital phalanx was a one day waiting period and lengthening at 1 mm/day. The mean healing indexes were 37.2 days/cm (range 24.2 to 41.5) in those with Apert syndrome and 64.3 days/cm in those with polydactyly (62.5 and 66.0). Our results suggest that osteogenesis at the distraction site may be quicker in patients with Apert syndrome than in those with polydactyly. PMID- 12564818 TI - Correction of congenital brachymetatarsia by gradual callus distraction. AB - Two children with congenital brachymetatarsia of the fourth metatarsal bone were treated by osteotomy of the metatarsal bone and bone lengthening by the Ilizarov technique. Both children were cosmetically improved. PMID- 12564819 TI - Simultaneous dislocation of the metacarpophalangeal and interphalangeal joints of the thumb. AB - Combined dislocation of the metacarpophalangeal and interphalangeal joints of the thumb is uncommon. We know of only four previously reported cases. We report a new case characterised by dorsal dislocation of both joints. Because of entrapment of the volar plate, open reduction at the interphalangeal joint was necessary. The metacarpophalangeal dislocation was treated by closed reduction. After three weeks of immobilisation, physiotherapy resulted in a satisfactory outcome. Even if the diagnosis of dislocation of the interphalangeal joint is obvious it would be easy to overlook a simultaneous dislocation of the metacarpophalangeal joint with serious consequences. Whole hand examination remains an essential rule. PMID- 12564820 TI - Chondroid syringoma of the hand. AB - We describe a patient with a benign chondroid syringoma of the little finger of the right hand. She was a 56-year-old pianist who had had the swelling for 25 years without it causing any symptoms. The tumour was excised with an excellent result. Although chondroid syringoma is rare, it should be included in the differential diagnosis of the soft tumours of the hand. PMID- 12564821 TI - Latina adolescents: predicting intentions to have sex. AB - This study utilized the theory of reasoned action to examine how Latina adolescents' intentions to have sex in the coming month were influenced by their general attitude toward having sex and their perceptions of general social norms about having sex. Eighty-four Mexican American and Central American adolescent females ages 14 to 19 participated in telephone interviews. One group of 41 Mexican American adolescents was obtained through a telephone survey and the second group of 43 pregnant or parenting Mexican American and Central American adolescents was recruited from health clinics. Path analysis conducted separately for the two groups indicated that, for the clinic group, perceptions of general social norms, but not general attitude, predicted intentions to have sex; and for the survey group, whether they were sexually active directly predicted perceptions of general social norms and intention to have sex. Adolescents' perceptions of what significant others desired of them, but not their motivation to comply, predicted perceptions of general social norms about having sex. Results underscore the importance of social norms for Latina adolescents when they are considering whether or not to have sex. PMID- 12564822 TI - Young dads: the effects of a parenting program on urban African-American adolescent fathers. AB - Sixty urban African-American adolescent first-time fathers were randomly assigned to two groups to study intervention strategies that would help them develop better and more consistent relationships with their young children. The fathers were administered a pretest interview schedule to determine their present quality of life as well as their relationships with their children. In addition to biweekly parenting classes, each member of the experimental group was assigned a social worker with whom he met weekly to assist him with his life needs. The control group was offered weekly parenting classes that focused on learning how to meet the infants' needs. At the end of six months, both groups were interviewed again. Findings indicated that the experimental group made significant gains in employment, vocational planning, feeling positive about their current relationships with their children, using birth control, being able to plan for the future, and increasing the number of close friends. PMID- 12564823 TI - Young women describe the ideal physician. AB - For some, the search for the ideal care provider can be elusive. This study explored female adolescents' accounts of the ideal health care provider. One hundred fifty-seven female high school students responded to the following question: "Can you describe what the ideal doctor would be like?" Content analysis of their descriptive narratives yielded 272 references to communication competence versus 30 references to medical competence (10 references were unrelated to either communication or medical competence). Based on their responses, it is clear that while young women appreciate the importance of medical skill, it is the communicatively competent care provider that they most seek. PMID- 12564824 TI - Personality characteristics as a function of frequency and type of substance use. AB - Problematic and unhealthy personality characteristics have typically been the focus of researchers interested in the possible causes of substance use/abuse. Recent research has begun to identify differences between individuals based upon frequency of use, with indications that moderate users may have "healthier" characteristics than either abstainers or regular users. The present study was conducted in a rural setting and compared personality structure (based on the California Psychological Inventory) among three groups of college students: abstainers, experimenters, and regular users. The comparisons were conducted as a function of all illicit substance use (excluding alcohol) and for alcohol use only. Minimal support was obtained for the above hypothesis when comparing the three groups on illicit substance use. When comparing the three groups on alcohol use only, support for the theory was identified, with fewer differences found between the experimenters and the abstainers than between the experimenters and the regular users. It is suggested that rural use of psychoactive substances may be more limited to alcohol due to easier availability and less stigma. Data supporting the "healthier" personality structure of moderate users of alcohol would thus be more likely in rural settings even though such use would still be "risky" behavior given its illegality for individuals under age 21. The need for adaptations to current treatment approaches is suggested. PMID- 12564825 TI - The effects of a mentoring program on at-risk youth. AB - This study examined an intensive mentoring program that focuses on youth deemed at-risk for juvenile delinquency or mental illness. Mothers and teachers completed the Child Behavior Checklist, and youth completed the Hopelessness Scale for Children, the Piers-Harris Self-Concept Scale, and the Self-Report Delinquency Scale. The youth (ages 10 to 17) either participated in the mentoring program (intervention, n = 34) or remained on the waiting list (nonintervention, n = 34) for 6 months. Repeated-measures ANOVAs assessed changes from preintervention to postintervention and indicated significant improvement in problematic behaviors for the intervention group. Mentoring appeared to affect African American youth differently than Caucasian and Latino youth. There were no significant interactions involving gender. The findings of this study supported the positive influence of mentoring on at-risk youth. PMID- 12564826 TI - Violence and touch deprivation in adolescents. AB - The increasing incidence of violence among children and adolescents highlights the importance of identifying at-risk profiles as well as assessing interventions for preventing violence. Empirical research has suggested behavioral, central nervous system, and neurotransmitter/neurohormone dysregulation in violent individuals, including (1) an underaroused central nervous system characterized by right frontal electroencephalogram (EEG) hypoactivation, and (2) a neurotransmitter/neurohormone profile of lower norepinephrine, serotonin, and cortisol, and elevated dopamine and testosterone. The literature also suggests a disproportionate incidence of physical abuse and neglect or the lack of positive physical contact in violent individuals. In the studies we have conducted to date, there has been a relatively high incidence of anger and aggression in high school samples, even those that were relatively advantaged, as well as high levels of depression (one standard deviation above the mean), suggesting significant disturbance in these youth. Adolescents with these profiles also had less optimal relationships with their families, used illicit drugs more frequently, had inferior academic performance, and had higher depression scores. In our cross-cultural comparisons, preschoolers and adolescents were less physically affectionate and more aggressive in the United States versus France. Further, the U.S. youth received less physical affection as preschoolers, and as adolescents they engaged in more self-stimulating behaviors, perhaps to compensate for receiving less physical affection from their parents and peers. This supports the notion that less physical affection (or more physical neglect) can contribute to greater aggression. Massage therapy has been effective with violent adolescents, perhaps because the physical stimulation reduced their dopamine levels and increased their serotonin levels. Their aggressive behavior decreased and their empathetic behavior increased. These preliminary data need to be replicated in a larger sample with a more comprehensive set of measures in the context of identifying a diagnostic profile. PMID- 12564827 TI - The role of ethnic identity and self-construal in coping among African American and Caucasian American seventh graders: an exploratory analysis of within-group variance. AB - Much coping research has been conducted comparing members of different ethnic groups using discrete racial classifications. However, the past two decades have seen the construct of ethnicity evolve into a complex variable that must be assessed in a more comprehensive way. This study explored how ethnicity, a discrete variable, and the continuous variables of a person's ethnic identity and self-construal contribute to the use of particular coping strategies across various situations. One hundred twelve seventh graders (67 African Americans and 45 Caucasian Americans) from three suburban middle schools completed questionnaires assessing ethnic identity, self-construal, and coping strategies for medical, test, and social criticism stressors. Results supported the hypothesis that ethnicity as a discrete variable is not associated with coping, but that ethnic identity and self-construal are. It was also found that high scores on the ethnic identity and self-construal scales were indicative of more positive psychological adjustment. Implications for future research and methodological considerations are discussed. PMID- 12564828 TI - Parent-child communication, perceived sanctions against drug use, and youth drug involvement. AB - To explore the relationship of parent-child communication to youth drug involvement, we analyzed secondary data from substance-abuse surveys given to 82,918 7th-12th graders in the U.S. It was found that: (a) parents are most often identified as the individuals who have talked to a child about drugs; (b) youth consider parents to be credible sources of information about drugs; (c) as perceived family sanctions go up, drug involvement goes down; (d) youth with the highest levels of drug involvement are the group most likely both to have had no one talk to them and to have had the largest number of people talk to them about drugs; and (e) perceived family sanctions increase dramatically for highly drug involved youth once they have been talked to by one person. PMID- 12564829 TI - Chinese adolescents' explanations of poverty: the Perceived Causes of Poverty Scale. AB - The Chinese Perceived Causes of Poverty Scale (CPCPS), constructed to assess how Chinese people explain poverty, covers four categories of explanations: personal problems of poor people, lack of opportunities to escape from the poverty cycle, exploitation of poor people, and bad fate. Chinese secondary school students (N = 1,519) were administered the CPCPS. Four factors were abstracted from their responses (Personal Problems, Lack of Opportunity, Exploitation, and Fate) and these factors (i.e., subscales) could reliably be reproduced in different subsamples. The four subscales were also found to be internally consistent and there was some support for their construct validity. PMID- 12564830 TI - Understanding adolescent suicide: a psychosocial interpretation of developmental and contextual factors. AB - Adolescent suicide remains an international tragedy, yet a common denominator continues to elude researchers. Some adolescents internalize rejection and respond with suicide; other troubled adolescents engage in homicide before ending their own lives. One factor underlying suicide concerns the failure to construct a healthy identity. Using Erikson's theories on identity development as a framework, this paper examines the motives for and contexts of suicide among preadolescents, adolescents, and young adults, identifies specific school-age populations that are vulnerable to suicide, and discusses implications. PMID- 12564831 TI - The identity status of African Americans in middle adolescence: a replication and extension of Forbes and Ashton (1998). AB - This study is a replication and extension of the work of Forbes and Ashton (1998). Seventy-seven African American high school students completed the revised version of the Extended Objective Measure of Ego Identity Status. Most of the students were found to be in uncommitted identity statuses. Similar results were found in both the ideological and interpersonal domains. PMID- 12564832 TI - The role of perceived social support and dysfunctional attitudes in predicting Taiwanese adolescents' depressive tendency. AB - This study examined the moderating effect of perceived social support on the association of dysfunctional attitudes with depression among Taiwanese adolescents. Seventh graders (N = 458) completed the Dysfunctional Attitudes Scale (Weisman & Beck, 1978), Perceived Social Support from Family and Friends (Procidano & Heller, 1983), Stressful Life Events (Chiu, 1988), and Children's Depression Inventory (Kovacs, 1981). The results indicated that perceived social support from friends moderates the relationship between dysfunctional attitudes and depression, implying that as peer support increases, the positive relationship between dysfunctional attitudes and depression weakens. PMID- 12564833 TI - An adolescent version of the Michigan Alcoholism Screening Test. AB - Item content of the Michigan Alcoholism Screening Test (MAST) was modified to make it more appropriate for young persons. The resulting test was found to have lower internal consistency than the adult MAST, but the elimination of five items with comparatively poor psychometric properties yielded an acceptable alpha coefficient of .73. A unitary factor model for the adolescent MAST was not confirmed; indeed, the revised test appeared to be factorially complex. Recommendations for further revisions are discussed. PMID- 12564834 TI - Personality disorders associated with substance use among American and Greek adolescents. AB - A study was conducted to investigate the association between personality disorder (PD) symptomatology and substance use among adolescents in community settings in the United States and Greece. The Structured Clinical Interview for DSM-IV Personality Disorders and the Adolescent Health Behavior Survey were completed by 37 male and 84 female adolescents, ages 15 to 18, who were recruited from an adolescent medical clinic and schools in and near New York City (n = 71) and Heraklion, Greece (n = 50). Results indicated that: (1) adolescents with PDs reported more frequent alcohol consumption during the past year than did those without PDs; (2) adolescents with borderline PD reported more cigarette smoking and heavy alcohol consumption than did those without borderline PD; (3) adolescents with antisocial PD symptomatology reported greater alcohol, cigarette, and illicit drug use than did those without antisocial PD symptomatology; and (4) although more American (30%) than Greek (4%) adolescents reported illicit substance use, differences were not observed in the prevalence of alcohol use, cigarette use, or personality disorders as a function of nationality. PMID- 12564835 TI - Combination of analytical quality specifications based on biological within- and between-subject variation. AB - At a conference on 'Strategies to Set Global Analytical Quality Specifications in Laboratory Medicine' in Stockholm 1999, a hierarchy of models to set analytical quality specifications was decided. The consensus agreement from the conference defined the highest level as 'evaluation of the effect of analytical performance on clinical outcomes in specific clinical settings' and the second level as 'data based on components of biological variation'. Here, the many proposals for analytical quality specifications based on biological variation are examined and the outcomes of the different models for maximum allowable combined analytical imprecision and bias are illustrated graphically. The following models were investigated. (1) The Cotlove et al. (1970) model defining analytical imprecision (%CVA) in relation to the within-subject biological variation (%CV(W-S)) as: %CVA < or = 0.5 x %CV(W-S) (where %CV is percentage coefficient of variation). (2) The Gowans et al. (1988) concept, which defines a functional relationship between analytical imprecision and bias for the maximum allowable combination of errors for the purpose of sharing common reference intervals. (3) The European Group for the Evaluation of Reagents and Analytical Systems in Laboratory Medicine (EGE Lab) Working Group concept, which combines the Cotlove model with the Gowans concept using the maximal acceptable bias. (4) The External Quality Assessment (EQA) Organizers Working Group concept, which is close to the EGE Lab Working Group concept, but follows the Gowans et al. concept of imprecision up to the limit defined by the model of Cotlove et al. (5) The 'three-level' concept classifying analytical quality into three levels: optimum, desirable and minimum. The figures created clearly demonstrated that the results obtained were determined by the basic assumptions made. When %CV(W-S) is small compared with the population-based coefficient of variation [%CV(P) = (%CV2(W-S) +%CV2(B S))(1/2)], the EGE Lab and EQA Organizers Working Group concepts become similar. Examples of analytical quality specifications based on biological variations are listed and an application on external quality control is illustrated for plasma creatinine. PMID- 12564836 TI - Receptor activator for nuclear factor kappaB ligand and osteoprotegerin: regulators of bone physiology and immune responses/potential therapeutic agents and biochemical markers. AB - The discovery of receptor activator for nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG) as the fundamental factors in controlling osteoclast formation and activation has led to a greater understanding of bone biology over the past few years. Here we discuss the role of these molecules in immunology and skeletal remodelling and assess their involvement in diseases of bones and joints, including rheumatoid arthritis, Paget's disease, post-menopausal osteoporosis and malignant bone diseases. OPG has been identified as a potential anabolic agent for treating conditions in which there is net bone loss and is currently in Phase I clinical trials. This review examines the current evidence indicating that OPG increases bone mass, and discusses other possible beneficial effects of OPG, such as inhibition of tumour growth and relief from bone cancer pain. OPG can be measured in human serum, and numerous studies have suggested that increased or decreased serum concentrations of this molecule can indicate the existence of remodelling disorders. Here we discuss how abnormal serum OPG concentrations could potentially be used to indicate imbalances of bone resorption and formation. The possible applications of serum OPG concentration as a marker for non-skeletal disease conditions are also considered. PMID- 12564837 TI - Phaeochromocytoma with normal urinary catecholamines: the potential value of urinary free metadrenalines. AB - BACKGROUND: Normal urine catecholamine values in patients with phaeochromocytoma is an occasional finding and may lead to a missed diagnosis. Additional urinary free metadrenaline analysis may be of value in this situation. METHODS: In addition to vanillylmandelic acid, homovanillic acid and the catecholamines, urinary free normetadrenaline (fNMA) and free metadrenaline (fMA) were measured. This report describes six confirmed cases of phaeochromocytoma showing normal urinary catecholamine output and compares fMA results and tumour size with other confirmed cases where the urine catecholamines were increased. RESULTS: Urine catecholamines in these patients with, on average, smaller tumours, were all normal. Urinary fNMA and fMA were available on five patients, and were increased in three. The data suggest that, unlike the catecholamines, urinary fNMA and fMA could be a useful predictor of tumour size. CONCLUSION: The inclusion of fNMA and fMA in the test profile is likely to be of additional benefit in tumour detection, particularly when catecholamines or other metabolites are normal. PMID- 12564838 TI - Temporal changes in maternal serum biochemical markers of trisomy 21 across the first and second trimester of pregnancy. AB - BACKGROUND: Many maternal serum markers show concentration changes in Down's syndrome pregnancies but the magnitude of the change in median marker levels varies with gestation. To date these changes have not been accurately specified. METHODS: The trends in marker median levels between 6 and 20 weeks of gestation were examined for alphafetoprotein (AFP), free beta human chorionic gonadotrophin (Fbeta-hCG), total human chorionic gonadotrophin (ThCG) and pregnancy-associated plasma protein A (PAPP-A) by a meta-analysis of data obtained from our collaborative studies and routine screening programmes for Down's syndrome over a 10-year period. Data were available from between 709 and 1082 Down's syndrome pregnancies and from between 14607 and 153909 unaffected pregnancies for each marker. The median multiple of the median (MoM) and mean log10MoM for each marker at each completed week of gestation were estimated and the trend with gestation smoothed using a weighted least squares regression model. RESULTS: The gestational ages at which maximum separation of marker levels occurred, comparing affected and unaffected pregnancies, and the respective regressed median MoMs and mean log10MoMs, were: for AFP at 16 weeks, 0.72 MoM, -0.14288log10MoM; for Fbeta hCG at 15 weeks, 2-24MoM, 0.35034 log10MoM; for ThCG at 16 weeks, 1.93 MoM, 0.28548 log10MoM, as well as before 8 weeks (<0.65 MoM, -0.18853 log10MoM); and for PAPP-A before 8 weeks, <0.33 MoM, -0.47727 log10MoM. CONCLUSION: There is significant temporal variation in mean log10MoM values for the screening markers investigated. Screening algorithms, modified to take account of this variation, should allow more accurate gestation-specific risks to be calculated in individual pregnancies. PMID- 12564839 TI - Evaluation of whole blood serotonin and plasma and urine 5-hydroxyindole acetic acid in diagnosis of carcinoid disease. AB - BACKGROUND: Carcinoid disease is an uncommon disorder resulting from tumours of the enterochromaffin cells. Current biochemical investigation usually involves the measurement of 5-hydroxyindole-3-acetic acid (5-HIAA) in 24-h urine collections. Because of the problems associated with urine collections (i.e. inconvenience, accuracy of collection and requirement for preservatives) two alternative markers, fasting plasma 5-HIAA and whole blood serotonin (5 hydroxytryptamine), have been studied. METHODS AND RESULTS: Whole blood serotonin concentration and plasma and urine 5-HIAA concentrations were measured by high performance liquid chromatography in 31 patients suspected of having carcinoid and 26 known carcinoid patients. Receiver operator characteristic curve analysis of the data showed no statistical difference between the three markers (P>0.01) with regard to their discriminating function. However, fasting plasma 5-HIAA assay showed greater stability than whole blood serotonin assay and is more convenient for the patient than a 24-h urine collection. At a cut-off value of 118 nmol/L plasma 5-HIAA assay showed a sensitivity of 89%, a specificity of 97% and a test efficiency of 93%. Whole blood serotonin assay was further limited by its saturation in platelets at 40 nmol/10(9) platelets which made it less suitable for monitoring the treatment of carcinoid disease. CONCLUSION: Fasting plasma 5-HIAA concentration provides a more convenient screening test for carcinoid and overcomes the problems associated with 24-h urine collections, without any loss of diagnostic precision. PMID- 12564840 TI - Influence of glucuronosyl bilirubin and (EZ)-cyclobilirubin on determination of serum unbound bilirubin by UB-analyser. AB - BACKGROUND: In the enzyme reaction for the determination of the unbound (free) bilirubin concentration by glucose oxidase and peroxidase, materials with low affinity for serum protein are reactive. The influence of these materials on the determination of serum unbound bilirubin was investigated. METHODS Serum samples from patients with neonatal hyperbilirubinaemia were analysed by high-performance liquid chromatography for total glucuronosyl bilirubin concentration (TGC) and (EZ)-cyclobilirubin concentration [(EZ)-C]. Based on these measurements, the samples were classified into three groups: group I [13 samples, TGC <2 micromol/L and (EZ)-C < 2.5 micromol/L]; group II [four samples, TGC < 2 micromol/L and (EZ) C > or = 2.5 micromol/L]; and group III (five samples, TGC > or = 2 micromol/L). The concentrations of total bilirubin and unbound bilirubin were measured in these same samples with a UB-analyser. When the absorbance at 460nm was monitored, the decrease in absorbance was non-linear (concave curve). The degree of concavity was estimated (D15 value) as the deviation from linearity at 15s. RESULTS: The D15 value was significantly higher in groups II and III than in group I. D15 value correlated significantly with TGC, (EZ)-C and unbound bilirubin concentration, and the unbound bilirubin concentration correlated significantly with TGC and (EZ)-C. CONCLUSION: These results indicated that determination of serum unbound bilirubin concentration using the UB-analyser could be positively skewed by high concentrations of TGC and (EZ)-C. PMID- 12564841 TI - Assay of catechol-O-methyltransferase activity in human erythrocytes using norepinephrine as a natural substrate. AB - BACKGROUND: Catechol-O-methyltransferase (COMT) catalyses the inactivation of catecholamines. It is widely distnbuted in most tissues in soluble (S-COMT) and membrane-bound (MB-COMT) forms. Recently, we used a new assay for COMT activity and demonstrated that COMT plays an important role in blood pressure regulation in spontaneously hypertensive rats. In order to investigate whether this is true for human hypertension, we have evaluated the erythrocyte COMT assay in humans. METHOD: The assay procedure included the use of norepinephrine (NE) as a natural substrate and the quantification of the reaction product, normetanephrine, followed by high-performance liquid chromatography separation and fluorescence or chemiluminescence detection. RESULTS: After evaluation of the method, the optimum conditions were obtained for the assay of human erythrocyte COMT. The S- and MB COMT activities obtained were 50.6 (24.5) and 329.8 (179.4) fmol/min/mg protein, respectively [mean (standard deviation); n = 54]. The Km values for NE were 91.3 (14.1) and 11.7 (1.1) micromol/L for S- and MB-COMT, respectively (n = 6). CONCLUSION: The established assay method used to assess S- and MB-COMT activities in human erythrocytes could be useful to elucidate catecholamine metabolism in the normal physiological state as well as in the pathology of certain diseases. PMID- 12564842 TI - Determination of D-amino acid oxidase activity in tumour cells. AB - BACKGROUND: We evaluated the assay for determining D-amino acid oxidase (DAAO) activity in tumour cells, rat liver and rat kidney for studying the effects of D amino acid-containing solution on cancer patients. METHODS AND RESULTS: In this method the amount of ammonia produced by the DAAO activity after removal of endogenous ammonia using a Sephadex G25 column was determined. The highest activity was observed in rat kidney, which was almost eight times that found in rat liver. As compared with host tissues, the DAAO activity in tumour cells was considerably less. CONCLUSIONS: This DAAO assay may be useful for analysis of various tissue samples as well as tumour cells. PMID- 12564843 TI - Creatinine assays: time for action? AB - BACKGROUND: Major differences in creatinine results between different laboratories and apparent inaccuracies when using commercial lyophilized standards were noted. In order to assess the variation and accuracy of the different methods we conducted a local audit. METHODS: To establish the variation between methods, plasma creatinine was measured on 47 human plasma samples by nine different laboratories using four different methods (Roche, Ortho, Olympus, modified Olympus). To establish the accuracy of the different methods, plasma creatinine was also determined on 16 of the plasma samples by tandem mass spectrometry (MS). In addition, all the laboratories measured the creatinine concentration on a commercial authenticated sample. RESULTS: All four methods gave significantly different (P<0.0001) plasma creatinine results when compared with each other. Generally, creatinine results produced by the Ortho method were considerably higher than those of the other methods, especially at higher creatinine concentrations (differences across methods between the lowest and highest result for the same sample ranged between 8% and 33%). All four methods generally gave higher results than those determined by tandem MS for samples with creatinine concentrations of < 250 micromol/L. Above this concentration the Olympus and Roche methods produced creatinine results that were lower then the tandem MS results, whereas results from the Ortho method were higher. Major matrix problems were found when a commercial lyophilized standard was used for creatnine estimation. CONCLUSION: No method gave good agreement with the tandem MS results, and there were major differences in measured plasma creatinine concentrations (up to 30% difference) between the various methods. We suggest that efforts should be made to standardize plasma creatinine measurement across all laboratories to minimize these problems. PMID- 12564844 TI - Detection of myelin basic protein in cerebrospinal fluid and serum from patients with HTLV-1-associated myelopathy/tropical spastic paraparesis. AB - BACKGROUND: Human T-cell lymphotropic virus type 1 (HTLV-1)-associated myelopathy (HAM)/tropical spastic paraparesis (TSP) is a chronic inflammatory disease that primarily affects the spinal cord and may be a neurological syndrome that is clinically similar to multiple sclerosis (MS). Myelin basic protein (MBP) in the cerebrospinal fluid (CSF) of MS patents is generally measured by radioimmunoassay. We have recently established a sensitive enzyme-linked immunosorbent assay (ELISA) and measured the MBP concentrations in CSF and serum of HAM/TSP patients. METHODS: A sensitive two-site ELISA capable of measuring MBP at a concentration as low as 30 pg/mL in serum and CSF samples was used. RESULTS: Significantly higher CSF MBP concentrations were detected in 61% of HAM/TSP patients than in patients with non-neurological diseases. Serum MBP concentrations were also higher in 9% of HAM/TSP patients compared with patients with non-neurological diseases or healthy controls. CONCLUSIONS: Using our ELISA system, we detected MBP in CSF and serum not only in patients with central active demyelination as in MS, but also in patients with spinal cord demyelination as in HAM/TSP. PMID- 12564845 TI - Serum sialic acid, a possible cardiovascular risk factor is not increased in Fijian Melanesians with impaired glucose tolerance or impaired fasting glucose. AB - BACKGROUND: Serum total sialic acid (TSA) has been shown to be a strong cardiovascular risk factor with increased concentrations being associated with increased mortality. Serum TSA is also elevated in patients with type 2 diabetes including those with micro- and macrovascular complications. We wished, therefore, to test the hypothesis that serum TSA may be abnormal in individuals with impaired glucose tolerance (IGT) and impaired fasting glucose (IFG), in Fijian Melanesians. METHOD: Twenty-one subjects with IGT (17 women and four men) were recruited along with 20 subjects with IFG (14 women and six men) and 22 normal subjects (12 women and 10 men). Serum TSA was 2.18 +/- 0.027 mmol//L, 2.19 +/- 0.033 mmol/L and 2.24 +/- 0-042 mmol/L in the three groups, respectively, which was not statistically different. Both systolic and diastolic blood pressure were, however, higher in the IGT group compared with the IFG and normal groups (P<0.04). CONCLUSION: Serum TSA is not elevated in Fijian Melanesians with IGT and IFG although it is reported to be elevated in type 2 diabetes mellitus in other populations. Further research is needed to establish why serum TSA is a potent independent cardiovascular risk factor and is elevated in type 2 diabetes mellitus in some populations. PMID- 12564846 TI - Ethnic differences in total and HDL cholesterol concentrations: Caucasians compared with predominantly Punjabi Sikh Indo-Asians. AB - BACKGROUND: In comparison with Caucasians, Indo-Asians resident in the UK have similar total cholesterol but lower HDL cholesterol (HDLC) concentrations. It is however possible that cardiovascular risk factors may vary between culturally different Indo-Asians. METHODS: We present data on 223 Indo-Asians (129 men, 94 women) and 787 Caucasians (421 men, 366 women) in whom a laboratory-based coronary heart disease (CHD) risk score calculation had been requested. RESULTS: Total cholesterol concentrations were similar in Indo-Asians and Caucasians. HDLC concentrations were higher (P < 0.001) in Caucasians [1.4 (1.3-1.4) mmol/L; median (95% confidence intervals)] than in Indo-Asians [1.2 (1.2-1.3) mmol/L]. Indo-Asian women [1.2 (1.2-1.3) mmol/L], Indo-Asian men [1.2 (1.2-1.3) mmol/L] and Caucasian men [1.2 (1.2-1.3) mmol/L] had similar HDLC concentrations but these were all lower (P < 0.001) than those in Caucasian women [1.4 (1.3-1.4) mmol/L]. CONCLUSION: We confirm low HDLC concentrations in Indo-Asians, but propose that this is solely due to low HDLC concentrations in Indo-Asian women. Since Indo-Asians in Wolverhampton are predominantly Punjabi Sikhs, we suggest that the difference between this study and previous reports may be due to heterogeneity of CHD risk factors within culturally diverse Indo-Asians. PMID- 12564847 TI - An improved quantitative assay of glycogen in erythrocytes. AB - BACKGROUND: Current methods for the quantitative determination of glycogen in erythrocytes are unsatisfactory. METHODS: Erythrocytes were deproteinized with perchloric acid either after haemolysing by freezing and thawing twice or without freezing and thawing, and the glycogen content was determined by an enzymatic method. RESULTS: Freezing and thawing resulted in a significantly higher glycogen content, and the recovery of added glycogen using this method was nearly 100%. The mean erythrocyte glycogen content in healthy volunteers (n = 17) was 69.5 microg/g haemoglobin, a value much higher than the previously reported values in healthy subjects. CONCLUSION: Freezing and thawing improves the assay of erythrocyte glycogen. PMID- 12564848 TI - Arterial blood gas analysis: selecting the clinically appropriate option for calculating base excess. AB - As part of arterial blood gas analysis, base excess is often reported as a measure of non-respiratory acid-base disturbance. Most blood gas analysers offer the option of calculating either the base excess of the blood sample or the base excess of the extracellular fluid (ECF). We report a case that illustrates that selecting the physiologically appropriate parameter avoids the potential for misinterpretation of acid-base data. We recommend that the base excess of the ECF is the appropriate metabolic blood gas parameter for clinical use. PMID- 12564849 TI - Hyperamylasaemia and multiple myeloma. AB - This case report describes a 42-year-old Caucasian woman who presented with persistent hyperamylasaemia and no evidence of pancreatic pathology. Further investigations resulted in a diagnosis of light-chain multiple myeloma. Amylase production by epithelial tumours has been well documented but the association with multiple myeloma has only been described in a small number of cases. The link does not appear to be immunoglobulin class-specific but the association with Bence Jones myeloma is unusual. The common features in this group of patients have been extensive extramedullary spread with a high tumour mass and a poor prognosis. This case was similar in that the patient showed very rapid disease activity developing extensive metastatic lesions and treatment ultimately proved unsuccessful. The amylase concentrations have been shown to decrease in response to treatment and increase at times of relapse and it has been proposed that it may be useful as a tumour marker in these patients. This case study adds to the pool of patents with this unusual association. PMID- 12564850 TI - Troponin for acute coronary syndromes. PMID- 12564851 TI - Transtheoretical model useful for patient education. PMID- 12564852 TI - Medical student participation valuable. PMID- 12564853 TI - The inhibition of stuttering: a viable alternative to contemporary therapy. PMID- 12564854 TI - Mnemonics helpful for physician-patient communication. PMID- 12564855 TI - In search of the roots of medicine: yesterday and today. PMID- 12564856 TI - The multiple mini-SOAP format for student presentations of complex patients. PMID- 12564857 TI - The summer assistantship in patient education: a preclinical preceptorship. AB - OBJECTIVES: The Swimmer Assistantship in Patient Education was developed to teach principles and techniques of brief preventive health counseling and patient education to medical students who have finished their first year. DESCRIPTION: Students complete an intensive training program and are then placed in family practice settings to educate and counsel patients full-time for 5 to 7 weeks. EVALUATION: Students evaluated the program after completion each year A survey of participants was conducted in fall 2000. CONCLUSIONS: Participants gained valuable experience with patient education and counseling techniques and a favorable experience in a family practice setting; many entered family practice residency training. PMID- 12564858 TI - The diaper change. PMID- 12564859 TI - The Seventh Chair: illuminating professional identity crises and ethical dilemmas in medicine. PMID- 12564860 TI - Geriatric medicine training for family practice residents in the 21st century: a report from the Residency Assistance Program/Harfford Geriatrics Initiative. AB - Increasing the quality and quantity of geriatric medicine training for family practice residents is a particular challenge for community-based programs. With support from the John A. Hartford Foundation of New York City, the American Academy of Family Physicians (AAFP) implemented in 1995 a multi-part project to improve the amount and quality of geriatric medicine education received by family practice residents. This report summarizes the initial results of the regional geriatric medicine curriculum retreats for residency directors. The goals of the retreats were to build recognition among the residency directors of the skills that future family physicians will require to be successful providers of primary care to older adults and to allow the residency directors to identify and develop solutions to barriers to improving geriatric medicine training for residents. Forty-six program directors participated in the three retreats between February 2000 and February 2001. The participants represented 52 programs and rural tracks in all geographic regions, small and large programs, and urban and rural settings. The program directors developed a consensus on the geriatric medicine knowledge, skills, and attitudes that should be expected of all family practice residency graduates; developed a list of basic, required educational resources for each family practice residency program; and proposed solutions to common obstacles to successful curriculum development. PMID- 12564861 TI - Geriatrics in family practice residency education: an unmet challenge. AB - The aging of the US population poses one of the greatest future challenges for family practice residency graduates. At a time when our discipline should be strengthening geriatric education to address the needs of our aging population, the Group on Geriatric Education of the Society of Teachers of Family Medicine believes that recent guidelines from important family medicine organizations suggest that our discipline's interest in geriatric education may be waning. Barriers to improving geriatric education in family practice residencies include limited geriatric faculty, changes in geriatric fellowship training, competing curricular demands, and limited diversity of geriatric training sites. Improving geriatric education in family practice residencies will require greater emphasis on faculty development and integration of geriatric principles throughout family practice residency education. The Residency Review Committee for Family Practice should review the Program Requirements for Residency Education to ensure that geriatric training requirements are consistent with current educational needs. The leadership of family medicine organizations should collaboratively address the need for continued improvement in training our residents to care for older patients and the chronically ill. PMID- 12564862 TI - A national survey on the current status of family practice residency education in geriatric medicine. AB - BACKGROUND AND OBJECTIVES: The dramatic increase in the elderly population expected over the next few decades will place a heavy strain on the current health care system. Family practice residents need to be prepared to take care of this geriatric population. In this study, we document the past, current, and future trends of geriatric education in family practice residency programs. METHODS: A survey was mailed to all family practice residency directors in the United States (n = 471). RESULTS: The response rate was 75%. Ninety-two percent of family practice residencies have a required geriatrics curriculum. Nursing homes, assisted living facilities, and home care are the predominant training sites for geriatrics. Training is most often offered in a longitudinal format. The mean number of physician faculty available to teach geriatrics is 2.6 per program (.83 full-time equivalent). Conflicting time demands with other curricula was ranked as the most significant barrier to geriatric education. Directors rated geriatrics as one of the three most important curriculum topics. CONCLUSIONS: Faculty development to enhance the number of faculty who can teach geriatrics and broadening the exposure of residents to the elderly in a variety of settings will be important to ensure that future generations of family physicians are adequately equipped to care for the geriatric population. PMID- 12564863 TI - Family medicine training in the care of older adults--has the retreat been sounded? PMID- 12564864 TI - Integrating population health into a family medicine clerkship: 7 years of evolution. AB - A population health curriculum using methodologies from community-oriented primary care (COPC) was developed in 1994 as part of a required third-year family medicine clerkship at the University of New Mexico. The curriculum integrates population health/community medicine projects and problem-based tutorials into a community-based, ambulatory clinical experience. By combining a required population health experience with relevant clinical training, student careers have the opportunity to be influenced during the critical third year. Results over a 7-year period describe a three-phase evolution of the curriculum, within the context of changes in medical education and in health care delivery systems in that same period of time. Early evaluation revealed that students viewed the curricular experience as time consuming and peripheral to their training. Later comments on the revised curriculum showed a higher regard for the experience that was described as important for student learning. PMID- 12564865 TI - Behavioral risks associated with tattooing. AB - BACKGROUND AND OBJECTIVES: Tattoos are an increasingly prevalent form of self expression, especially for adolescents. This study was conducted to determine health-risk behaviors associated with tattoos in young men and women entering military service. METHODS: We surveyed a cohort of 550 military recruits using a modification of the Youth Risk Behavioral Survey (YRBS), a validated instrument used to assess health risk behaviors in adolescents. All individuals entering basic training in the US Marine Corps or the US Air Force from June through September 1999 were eligible to participate. The primary outcome variables of interest were tobacco use, alcohol use, seatbelt use, suicidal behaviors, depression, and physical violence. RESULTS: The survey response rate was 91% (n = 499 of 550). Overall, 27% of respondents had tattoos (n = 125) when entering military service. Women entering military service were more likely to have a tattoo than men. Controlling for age and gender, individuals with tattoos were more likely to smoke, drink heavily, use smokeless tobacco, and ride in a vehicle with someone who had been drinking than non-tattooed individuals. CONCLUSIONS: In a population of military recruits, tattoos were associated with predictable adverse health-risk behaviors. This represents an important opportunity for targeted preventive counseling. PMID- 12564866 TI - Introducing personal digital assistants to family physician teachers. AB - BACKGROUND AND OBJECTIVES: In our previous projects, students and residents have readily adopted personal digital assistants (PDAs), but faculty have generally been reluctant. The objective of the project reported here was to maximize adoption of PDAs by our faculty, using a combination of strategies. METHODS: Through cost-shared funding, we provided full-time and community teachers with PocketPCs or Handspring Visors, along with preinstalled medical software. Use patterns and satisfaction were assessed by structured questionnaire and focus group discussions. RESULTS: For the calendar, address book, and pharmacopoeia, we found that 83% of faculty use these two to three times per day. Cost sharing and software preinstallation were popular. Device synchronization and e-mail showed potential but caused problems. Easy access to technical support from peers and a variety of information-sharing structures eased maintenance issues. Point-of-care data access was important to faculty. CONCLUSIONS: With the right support structures, faculty adopt PDAs in clinical and teaching settings. PMID- 12564868 TI - Three-dimensional cardiovascular image analysis. PMID- 12564867 TI - Cervical cancer rates and the supply of primary care physicians in Florida. AB - BACKGROUND AND OBJECTIVES: This study's aim was to determine if an increased supply of primary care physicians is associated with lower incidence and mortality rates for cervical cancer. METHODS: We determined cervical cancer incidence and mortality rates for each of Florida's 67 counties over the 3-year period of 1993-1995 using data from Florida's population-based tumor registry. Data on physician supply were obtained from the 1994 American Medical Association Physician Masterfile. We used multiple linear regression analysis to examine the relationship between physician supply and cervical cancer incidence and mortality rates, adjusting for other county-level characteristics. RESULTS: In regression analysis that adjusted for other county-level characteristics, each increase in the supply of family physicians of one physician/10,000 persons was associated with a corresponding drop in the incidence rate of 1.5 cases/100,000 persons and a corresponding drop in mortality rate of .65 cases/100,000 persons. CONCLUSIONS: Our results indicate that a greater supply of primary care physicians is likely associated with a lower incidence of cervical cancer and a lower cervical cancer mortality rate. More studies are needed at the individual patient level to confirm this association. PMID- 12564869 TI - A review of cardiac image registration methods. AB - In this paper, the current status of cardiac image registration methods is reviewed. The combination of information from multiple cardiac image modalities, such as magnetic resonance imaging, computed tomography, positron emission tomography, single-photon emission computed tomography, and ultrasound, is of increasing interest in the medical community for physiologic understanding and diagnostic purposes. Registration of cardiac images is a more complex problem than brain image registration because the heart is a nonrigid moving organ inside a moving body. Moreover, as compared to the registration of brain images, the heart exhibits much fewer accurate anatomical landmarks. In a clinical context, physicians often mentally integrate image information from different modalities. Automatic registration, based on computer programs, might, however, offer better accuracy and repeatability and save time. PMID- 12564870 TI - Echocardiographic strain and strain-rate imaging: a new tool to study regional myocardial function. AB - Ultrasonic imaging is the noninvasive clinical imaging modality of choice for diagnosing heart disease. At present, two-dimensional ultrasonic grayscale images provide a relatively cheap, fast, bedside method to study the morphology of the heart. Several methods have been proposed to assess myocardial function. These have been based on either grayscale or motion (velocity) information measured in real-time. However, the quantitative assessment of regional myocardial function remains an important goal in clinical cardiology. To do this, ultrasonic strain and strain-rate imaging have been introduced. In the clinical setting, these techniques currently only allow one component of the true three-dimensional deformation to be measured. Clinical, multidimensional strain (rate) information can currently thus only be obtained by combining data acquired using different transducer positions. Nevertheless, given the appropriate postprocessing, the clinical value of these techniques has already been shown. Moreover, multidimensional strain and strain-rate estimation of the heart in vivo by means of a single ultrasound acquisition has been shown to be feasible. In this paper, the new techniques of ultrasonic strain rate and strain imaging of the heart are reviewed in terms of definitions, data acquisition, strain-rate estimation, postprocessing, and parameter extraction. Their clinical validation and relevance will be discussed using clinical examples on relevant cardiac pathology. Based on these examples, suggestions are made for future developments of these techniques. PMID- 12564871 TI - Model-based imaging of cardiac electrical excitation in humans. AB - Activation time (AT) imaging from electrocardiographic (ECG) mapping data has been developing for several years. By coupling ECG mapping and three-dimensional (3-D) + time anatomical data, the electrical excitation sequence can be imaged completely noninvasively in the human heart. In this paper, a bidomain theory based surface heart model AT imaging approach was applied to single-beat data of atrial and ventricular depolarization in two patients with structurally normal hearts. In both patients, the AT map was reconstructed from sinus and paced rhythm data. Pacing sites were the apex of the right ventricle and the coronary sinus (CS) ostium. For CS pacing, the reconstructed AT pattern on the endocardium of the right atrium was compared with the CARTO map in both patients. The localization errors of the origins of the initial endocardial breakthroughs were determined to be 6 and 12 mm. The sites of early activation and the areas with late activation were estimated with sufficient accuracy. The reconstructed sinus rhythm sequence was in good qualitative agreement with the pattern previously published for the isolated Langendorff-perfused human heart. PMID- 12564872 TI - Application of task-based measures of image quality to optimization and evaluation of three-dimensional reconstruction-based compensation methods in myocardial perfusion SPECT. AB - In this paper, we apply the channelized Hotelling observer (CHO) using a defect detection task to the optimization and evaluation of three-dimensional iterative reconstruction-based compensation methods for myocardial perfusion single-photon emission computed tomography (SPECT). We used a population of 24 mathematical cardiac-torso phantoms that realistically model the activity and attenuation distribution in three classes of patients: females, and males with flat diaphragms and raised diaphragms. Projection data were generated and subsequently reconstructed using methods based on the ordered subsets-expectation maximization (OSEM) algorithm. The methods evaluated included compensation for attenuation, detector response blurring, and scatter in various combinations. We applied the CHO to optimize the number of iterations for OSEM and the cutoff frequency and order of a three-dimensional postreconstruction Butterworth filter. Using the optimal parameters, we then compared the compensation methods. The index of comparison in these studies was the area under the receiver operating characteristics curve (AUC) for the CHO. We found that attenuation compensation with either detector response or scatter compensation gave statistically significant increases in the AUC compared to attenuation compensation alone. The greatest increase in the AUC occurred when all three compensations were applied. These results indicate that compensation for detector response and scatter, in addition to attenuation compensation, will improve defect detectability in myocardial SPECT images. PMID- 12564873 TI - 3-D freehand echocardiography for automatic left ventricle reconstruction and analysis based on multiple acoustic windows. AB - A new method is proposed to reconstruct and analyze the left ventricle (LV) from multiple acoustic window three-dimensional (3-D) ultrasound acquired using a transthoracic 3-D rotational probe. Prior research in this area has been based on one acoustic window acquisition. However, the data suffers from several limitations that degrade the reconstruction and reduce the clinical value of interpretation, such as the presence of shadow due to bone (ribs) and air (in the lungs) and motion of the probe during the acquisition. In this paper, we show how to overcome these limitations by automatically fusing information from multiple acoustic window sparse-view acquisitions and using a position sensor to track the probe in real time. Geometric constraints of the object shape, and spatiotemporal information relating to the image acquisition process, are used in new algorithms for 1) grouping endocardial edge cues from an initial image segmentation and 2) defining a novel reconstruction method that utilizes information from multiple acoustic windows. The new method has been validated on a phantom and three real heart data sets. In the phantom study, one finger of a latex glove was scanned from two acoustic windows and reconstructed using the new method. The volume error was measured to be less than 4%. In the clinical case study, 3-D ultrasound and magnetic resonance imaging (MRI) scanning were performed on the same healthy volunteers. Quantitative ejection fractions (EFs) and volume-time curves over a cardiac cycle were estimated using the new method and compared to cardiac MRI measurements. This showed that the new method agrees better with MRI measurements than the previous approach we have developed based on a single acoustic window. The EF errors of the new method with respect to MRI measurements were less than 6%. A more extensive clinical validation is required to establish whether these promising first results translate to a method suitable for routine clinical use. PMID- 12564874 TI - Efficient model-based quantification of left ventricular function in 3-D echocardiography. AB - Quantitative functional analysis of the left ventricle plays a very important role in the diagnosis of heart diseases. While in standard two-dimensional echocardiography this quantification is limited to rather crude volume estimation, three-dimensional (3-D) echocardiography not only significantly improves its accuracy but also makes it possible to derive valuable additional information, like various wall-motion measurements. In this paper, we present a new efficient method for the functional evaluation of the left ventricle from 3-D echographic sequences. It comprises a segmentation step that is based on the integration of 3-D deformable surfaces and a four-dimensional statistical heart motion model. The segmentation results in an accurate 3-D + time left ventricle discrete representation. Functional descriptors like local wall-motion indexes are automatically derived from this representation. The method has been successfully tested both on electrocardiography-gated and real-time 3-D data. It has proven to be fast, accurate, and robust. PMID- 12564875 TI - Automated 3-D echocardiography analysis compared with manual delineations and SPECT MUGA. AB - A major barrier for using 3-D echocardiography for quantitative analysis of heart function in routine clinical practice is the absence of accurate and robust segmentation and tracking methods necessary to make the analysis automatic. In this paper, we present an automated three-dimensional (3-D) echocardiographic acquisition and image-processing methodology for assessment of left ventricular (LV) function. We combine global image information provided by a novel multiscale fuzzy-clustering segmentation algorithm, with local boundaries obtained with phase-based acoustic feature detection. We then use the segmentation results to fit and track the LV endocardial surface using a 3-D continuous transformation. To our knowledge, this is the first report of a completely automated method. The protocol is evaluated in a small clinical case study (nine patients). We compare ejection fractions (EFs) computed with the new approach to those obtained using the standard clinical technique, single-photon emission computed tomography multigated acquisition. Errors on six datasets were found to be within six percentage points. A further two, with poor image quality, improved upon EFs from manually delineated contours, and the last failed due to artifacts in the data. Volume-time curves were derived and the results compared to those from manual segmentation. Improvement over an earlier published version of the method is noted. PMID- 12564876 TI - Integrated surface model optimization for freehand three-dimensional echocardiography. AB - The major obstacle of three-dimensional (3-D) echocardiography is that the ultrasound image quality is too low to reliably detect features locally. Almost all available surface-finding algorithms depend on decent quality boundaries to get satisfactory surface models. We formulate the surface model optimization problem in a Bayesian framework, such that the inference made about a surface model is based on the integration of both the low-level image evidence and the high-level prior shape knowledge through a pixel class prediction mechanism. We model the probability of pixel classes instead of making explicit decisions about them. Therefore, we avoid the unreliable edge detection or image segmentation problem and the pixel correspondence problem. An optimal surface model best explains the observed images such that the posterior probability of the surface model for the observed images is maximized. The pixel feature vector as the image evidence includes several parameters such as the smoothed grayscale value and the minimal second directional derivative. Statistically, we describe the feature vector by the pixel appearance probability model obtained by a nonparametric optimal quantization technique. Qualitatively, we display the imaging plane intersections of the optimized surface models together with those of the ground truth surfaces reconstructed from manual delineations. Quantitatively, we measure the projection distance error between the optimized and the ground-truth surfaces. In our experiment, we use 20 studies to obtain the probability models offline. The prior shape knowledge is represented by a catalog of 86 left ventricle surface models. In another set of 25 test studies, the average epicardial and endocardial surface projection distance errors are 3.2 +/- 0.85 mm and 2.6 +/- 0.78 mm, respectively. PMID- 12564877 TI - ROPES: a semiautomated segmentation method for accelerated analysis of three dimensional echocardiographic data. AB - Echocardiography (cardiac ultrasound) is today the predominant technique for quantitative assessment of cardiac function and valvular heart lesions. Segmentation of cardiac structures is required to determine many important diagnostic parameters. As the heart is a moving organ, reliable information can be obtained only from three-dimensional (3-D) data over time (3-D + time = 4-D). Due to their size, the resulting four-dimensional (4-D) data sets are not reasonably accessible to simple manual segmentation methods. Automatic segmentation often yields unsatisfactory results in a clinical environment, especially for ultrasonic images. We describe a semiautomated segmentation algorithm (ROPES) that is able to greatly reduce the time necessary for user interaction and its application to extract various parameters from 4-D echocardiographic data. After searching for candidate contour points, which have to fulfill a multiscale edge criterion, the candidates are connected by minimizing a cost function to line segments that then are connected to form a closed contour. The contour is automatically checked for plausibility. If necessary, two correction methods that can also be used interactively are applied (fitting of other line segments into the contour and searching for additional candidates with a relaxed criterion). The method is validated using in vivo transesophageal echocardiographic data sets. PMID- 12564878 TI - Kinematics of the heart: strain-rate imaging from time-resolved three-dimensional phase contrast MRI. AB - A four-dimensional mapping (three spatial dimensions + time) of myocardial strain rate would help to describe the mechanical properties of the myocardium, which affect important physiological factors such as the pumping performance of the ventricles. Strain-rate represents the local instantaneous deformation of the myocardium and can be calculated from the spatial gradients of the velocity field. Strain-rate has previously been calculated using one-dimensional (ultrasound) or two-dimensional (2-D) magnetic resonance imaging) techniques. However, this assumes that myocardial motion only occurs in one direction or in one plane, respectively. This paper presents a method for calculation of the time resolved three-dimensional (3-D) strain-rate tensor using velocity vector information in a 3-D spatial grid during the whole cardiac cycle. The strain-rate tensor provides full information of both magnitude and direction of the instantaneous deformation of the myocardium. A method for visualization of the full 3-D tensor is also suggested. The tensors are visualized using ellipsoids, which display the principal directions of strain-rate and the ratio between strain-rate magnitude in each direction. The presented method reveals the principal strain-rate directions without a priori knowledge of myocardial motion directions. PMID- 12564879 TI - A MAP framework for tag line detection in SPAMM data using Markov random fields on the B-spline solid. AB - Magnetic resonance (MR) tagging is a technique for measuring heart deformations through creation of a stripe grid pattern on cardiac images. In this paper, we present a maximum a posteriori (MAP) framework for detecting tag lines using a Markov random field (MRF) defined on the lattice generated by three-dimensional (3-D) and four-dimensional (4-D) (3-D + t) uniform sampling of B-spline models. In the 3-D case, MAP estimation is cast for detecting present tag features in the current image given an initial solid from the previous frame (the initial undeformed solid is manually positioned by clicking on corner points of a cube). The method also allows the parameters of the solid model, including the number of knots and the spline order, to be adjusted within the same framework. Fitting can start with a solid with less knots and lower spline order and proceed to one with more knots and/or higher order so as to achieve more accuracy and/or higher order of smoothness. In the 4-D case, the initial model is considered to be the linear interpolation of a sequence of optimal solids obtained from 3-D tracking. The same framework proposed for the 3-D case can once again be applied to arrive at a 4-D B-spline model with a higher temporal order. PMID- 12564880 TI - Virtual tagging: numerical considerations and phantom validation. AB - This paper presents a virtual tagging framework for measuring, as well as visualising, myocardial deformation using magnetic resonance (MR) velocity imaging. Tagging grids are allocated artificially according to the deformation gradient with varying shapes and densities. The control points are then deformed such that the difference between the induced deformation velocity and that of actually measured MR data is minimum. A full three-dimensional implementation of the technique combined with the mass conservation constraint is provided. Numerical considerations of applying the proposed framework and different optimization strategies have been investigated with both simulated and phantom experiments. The accuracy of the technique in terms of following material deformation is compared with that of conventional tagging technique. PMID- 12564881 TI - Model evaluation and calibration for prospective respiratory motion correction in coronary MR angiography based on 3-D image registration. AB - Image processing was used as a fundamental tool to derive motion information from magnetic resonance (MR) images, which was fed back into prospective respiratory motion correction during subsequent data acquisition to improve image quality in coronary MR angiography (CMRA) scans. This reduces motion artifacts in the images and, in addition, enables the usage of a broader gating window than commonly used today to increase the scan efficiency. The aim of the study reported in this paper was to find a suitable motion model to be used for respiratory motion correction in cardiac imaging and to develop a calibration procedure to adapt the motion model to the individual patient. At first, the performance of three motion models [one-dimensional translation in feet-head (FH) direction, three dimensional (3-D) translation, and 3-D affine transformation] was tested in a small volunteer study. An elastic image registration algorithm was applied to 3-D MR images of the coronary vessels obtained at different respiratory levels. A strong intersubject variability was observed. The 3-D translation and affine transformation model were found to be superior over the conventional FH translation model used today. Furthermore, a new approach is presented, which utilizes a fast model-based image registration to extract motion information from time series of low-resolution 3-D MR images, which reflects the respiratory motion of the heart. The registration is based on a selectable global 3-D motion model (translation, rigid, or affine transformation). All 3-D MR images were registered with respect to end expiration. The resulting time series of model parameters were analyzed in combination with additionally acquired motion information from a diaphragmatic MR pencil-beam navigator to calibrate the respiratory motion model. To demonstrate the potential of a calibrated motion model for prospective motion correction in coronary imaging, the approach was tested in CMRA examinations in five volunteers. PMID- 12564882 TI - A study of the motion and deformation of the heart due to respiration. AB - This paper describes a quantitative assessment of respiratory motion of the heart and the construction of a model of respiratory motion. Three-dimensional magnetic resonance scans were acquired on eight normal volunteers and ten patients. The volunteers were imaged at multiple positions in the breathing cycle between full exhalation and full inhalation while holding their breath. The exhalation volume was segmented and used as a template to which the other volumes were registered using an intensity-based rigid registration algorithm followed by nonrigid registration. The patients were imaged at inhale and exhale only. The registration results were validated by visual assessment and consistency measurements indicating subvoxel registration accuracy. For all subjects, we assessed the nonrigid motion of the heart at the right coronary artery, right atrium, and left ventricle. We show that the rigid-body motion of the heart is primarily in the craniocaudal direction with smaller displacements in the right left and anterior-posterior directions; this is in agreement with previous studies. Deformation was greatest for the free wall of the right atrium and the left ventricle; typical deformations were 3-4 mm with deformations of up to 7 mm observed in some subjects. Using the registration results, landmarks on the template surface were mapped to their correct positions through the breathing cycle. Principal component analysis produced a statistical model of the motion and deformation of the heart. We discuss how this model could be used to assist motion correction. PMID- 12564883 TI - Automatic construction of multiple-object three-dimensional statistical shape models: application to cardiac modeling. AB - A novel method is introduced for the generation of landmarks for three dimensional (3-D) shapes and the construction of the corresponding 3-D statistical shape models. Automatic landmarking of a set of manual segmentations from a class of shapes is achieved by 1) construction of an atlas of the class, 2) automatic extraction of the landmarks from the atlas, and 3) subsequent propagation of these landmarks to each example shape via a volumetric nonrigid registration technique using multiresolution B-spline deformations. This approach presents some advantages over previously published methods: it can treat multiple part structures and requires less restrictive assumptions on the structure's topology. In this paper, we address the problem of building a 3-D statistical shape model of the left and right ventricle of the heart from 3-D magnetic resonance images. The average accuracy in landmark propagation is shown to be below 2.2 mm. This application demonstrates the robustness and accuracy of the method in the presence of large shape variability and multiple objects. PMID- 12564884 TI - 3-D active appearance models: segmentation of cardiac MR and ultrasound images. AB - A model-based method for three-dimensional image segmentation was developed and its performance assessed in segmentation of volumetric cardiac magnetic resonance (MR) images and echocardiographic temporal image sequences. Comprehensive design of a three-dimensional (3-D) active appearance model (AAM) is reported for the first time as an involved extension of the AAM framework introduced by Cootes et al. The model's behavior is learned from manually traced segmentation examples during an automated training stage. Information about shape and image appearance of the cardiac structures is contained in a single model. This ensures a spatially and/or temporally consistent segmentation of three-dimensional cardiac images. The clinical potential of the 3-D AAM is demonstrated in short-axis cardiac MR images and four-chamber echocardiographic sequences. The method's performance was assessed by comparison with manually identified independent standards in 56 clinical MR and 64 clinical echo image sequences. The AAM method showed good agreement with the independent standard using quantitative indexes of border positioning errors, endo- and epicardial volumes, and left ventricular mass. In MR, the endocardial volumes, epicardial volumes, and left ventricular wall mass correlation coefficients between manual and AAM were R2 = 0.94, 0.97, 0.82, respectively. For echocardiographic analysis, the area correlation was R2 = 0.79. The AAM method shows high promise for successful application to MR and echocardiographic image analysis in a clinical setting. PMID- 12564885 TI - Four-dimensional wavelet compression of arbitrarily sized echocardiographic data. AB - Wavelet-based methods have become most popular for the compression of two dimensional medical images and sequences. The standard implementations consider data sizes that are powers of two. There is also a large body of literature treating issues such as the choice of the "optimal" wavelets and the performance comparison of competing algorithms. With the advent of telemedicine, there is a strong incentive to extend these techniques to higher dimensional data such as dynamic three-dimensional (3-D) echocardiography [four-dimensional (4-D) datasets]. One of the practical difficulties is that the size of this data is often not a multiple of a power of two, which can lead to increased computational complexity and impaired compression power. Our contribution in this paper is to present a genuine 4-D extension of the well-known zerotree algorithm for arbitrarily sized data. The key component of our method is a one-dimensional wavelet algorithm that can handle arbitrarily sized input signals. The method uses a pair of symmetric/antisymmetric wavelets (10/6) together with some appropriate midpoint symmetry boundary conditions that reduce border artifacts. The zerotree structure is also adapted so that it can accommodate noneven data splitting. We have applied our method to the compression of real 3-D dynamic sequences from clinical cardiac ultrasound examinations. Our new algorithm compares very favorably with other more ad hoc adaptations (image extension and tiling) of the standard powers-of-two methods, in terms of both compression performance and computational cost. It is vastly superior to slice-by-slice wavelet encoding. This was seen not only in numerical image quality parameters but also in expert ratings, where significant improvement using the new approach could be documented. Our validation experiments show that one can safely compress 4-D data sets at ratios of 128:1 without compromising the diagnostic value of the images. We also display some more extreme compression results at ratios of 2000:1 where some key diagnostically relevant key features are preserved. PMID- 12564886 TI - Predictive (un)distortion model and 3-D reconstruction by biplane snakes. AB - This paper is concerned with the three-dimensional (3-D) reconstruction of coronary vessel centerlines and with how distortion of X-ray angiographic images affects it. Angiographies suffer from pincushion and other geometrical distortions, caused by the peripheral concavity of the image intensifier (II) and the nonlinearity of electronic acquisition devices. In routine clinical practice, where a field-of-view (FOV) of 17-23 cm is commonly used for the acquisition of coronary vessels, this distortion introduces a positional error of up to 7 pixels for an image matrix size of 512 x 512 and an FOV of 17 cm. This error increases with the size of the FOV. Geometrical distortions have a significant effect on the validity of the 3-D reconstruction of vessels from these images. We show how this effect can be reduced by integrating a predictive model of (un)distortion into the biplane snakes formulation for 3-D reconstruction. First, we prove that the distortion can be accurately modeled using a polynomial for each view. Also, we show that the estimated polynomial is independent of focal length, but not of changes in anatomical angles, as the II is influenced by the earth's magnetic field. Thus, we decompose the polynomial into two components: the steady and the orientation-dependent component. We determine the optimal polynomial degree for each component, which is empirically determined to be five for the steady component and three for the orientation-dependent component. This fact simplifies the prediction of the orientation-dependent polynomial, since the number of polynomial coefficients to be predicted is lower. The integration of this model into the biplane snakes formulation enables us to avoid image unwarping, which deteriorates image quality and therefore complicates vessel centerline feature extraction. Moreover, we improve the biplane snake behavior when dealing with wavy vessels, by means of using generalized gradient vector flow. Our experiments show that the proposed methods in this paper decrease up to 88% the reconstruction error obtained when geometrical distortion effects are ignored. Tests on imaged phantoms and real cardiac images are presented as well. PMID- 12564887 TI - Left ventricular volume estimation for real-time three-dimensional echocardiography. AB - The application of level set techniques to echocardiographic data is presented. This method allows semiautomatic segmentation of heart chambers, which regularizes the shapes and improves edge fidelity, especially in the presence of gaps, as is common in ultrasound data. The task of the study was to reconstruct left ventricular shape and to evaluate left ventricular volume. Data were acquired with a real-time three-dimensional (3-D) echocardiographic system. The method was applied directly in the three-dimensional domain and was based on a geometric-driven scheme. The numerical scheme for solving the proposed partial differential equation is borrowed from numerical methods for conservation law. Results refer to in vitro and human in vivo acquired 3-D + time echocardiographic data. Quantitative validation was performed on in vitro balloon phantoms. Clinical application of this segmentation technique is reported for 20 patient cases providing measures of left ventricular volumes and ejection fraction. PMID- 12564888 TI - Environmental costs of freshwater eutrophication in England and Wales. AB - Eutrophication has many known consequences, but there are few data on the environmental and health costs. We developed a new framework of cost categories that assess both social and ecological damage costs and policy response costs. These findings indicate the severe effects of nutrient enrichment and eutrophication on many sectors of the economy. We estimate the damage costs of freshwater eutrophication in England and Wales to be $105-160 million yr(-1) (pound 75.0-114.3 m). The policy response costs are a measure of how much is being spent to address this damage, and these amount to $77 million yr(-1) pound 54.8 m). The damage costs are dominated by seven items each with costs of $15 million yr(-1) or more: reduced value of waterfront dwellings, drinking water treatment costs for nitrogen removal, reduced recreational and amenity value of water bodies, drinking water treatment costs for removal of algal toxins and decomposition products, reduced value of nonpolluted atmosphere, negative ecological effects on biota, and net economic losses from the tourist industry. In common with other environmental problems, it would represent net value (or cost reduction) if damage was prevented at source. A variety of effective economic, regulatory, and administrative policy instruments are available for internalizing these costs. PMID- 12564889 TI - Orographic cold-trapping of persistent organic pollutants by vegetation in mountains of western Canada. AB - Conifer needles from mountain areas of Alberta and British Columbia, Canada, were collected from sites that ranged in altitude from 770 to 2200 masl and were analyzed for polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCs) to determine if they are progressively concentrated in colder, more elevated mountain areas, where temperatures decrease as elevation increases. Concentrations of OCs in needles ranged from 43 to 2430 pg g(-1), 55-17500 pg g( 1), and 11-2930 pg g(-1) (dry weight), for total hexachlorocyclohexanes (HCHs), PCBs, and endosulfans, respectively. The more volatile OCs, with subcooled liquid vapor pressures (PL) > 0.1 Pa at 25 degrees C, increased at higher altitudes, whereas the less volatile OCs were either unrelated or inversely correlated with altitude. These spatial patterns were similar for species of spruce (Picea engelmannii and glauca) and pine (Pinus contorta and albicaulis). Back trajectories revealed that air masses arriving at these sites traveled over Asia and the Pacific Ocean before reaching the Rocky Mountains. Results from this study demonstrate that alpine ecosystems accumulate these chemicals to the same degree that is observed in polar environments that are known to receive contaminants by long-range transport. PMID- 12564890 TI - Events affecting levels and seasonal evolution of airborne particulate matter concentrations in the Western Mediterranean. AB - Time series (1996-2000) of levels of PM (PM10 and TSP) and gaseous pollutants recorded in air quality monitoring networks from Northeastern Spain were interpreted using meteorological data and satellite observations of African dust plumes. The main objective of this study was to identify the processes affecting time variations of PM levels on a day-to-day and seasonal basis. From March to October PM levels at rural, urban, and industrial sites vary as a function of the concatenation of Atlantic air mass advections (Atlantic episodes with low PM levels) and regional circulations (regional events with high PM levels, very often associated with high ozone levels), which favor the aging of air masses in the Western Mediterranean basin. During these regional episodes, PM is transported from urban/industrial to rural sites by meso-scale circulations. From November to February low PM levels are recorded at rural sites, and variations in PM levels at urban/industrial sites are governed by the successive occurrence of Atlantic episodes and local urban/industrial pollution events. The African dust outbreaks take place throughout the year and may induce PM levels to increase simultaneously in large areas of the Iberian peninsula. The difference between PM concentrations measured at urban and rural sites experiences a seasonal trend similar to that of levels of NO(x) and CO, which is characterized by a winter maximum due to the higher frequency of intensive local urban pollution events. However, maximum PM levels are recorded in summer at rural sites owing to the frequent occurrence of regional episodes. Furthermore, in three years of the study period (1997, 1998, and 2000), a second-order PM maximum was also recorded at rural sites in March owing to intensive African dust outbreaks. PMID- 12564891 TI - Effects of historic PCB exposures on the reproductive success of the Hudson River striped bass population. AB - Scientists and regulatory agencies have expressed concern that exposures to polychlorinated biphenyls (PCBs) might be contributing to reductions in the abundance of fish populations exposed to these chemicals. The specific effects of concern involve impairment of fish reproduction, including both reduced egg production and decreased viability of eggs and larvae. We tested hypotheses concerning the effects of PCBs on fish populations using long-term data sets available for the striped bass population of the Hudson River, NY, a population that has long been a subject of regulatory concern because of potential effects of PCB exposures. The data sets examined include both measurements of PCB concentrations in adult female striped bass over the period from 1976 through 1997 and estimates of the numbers of striped bass eggs, larvae, and juveniles produced annually during this same period. We found strong correlations between estimates of the abundance of spawners and the number of eggs and larvae produced by those spawners and also between independent estimates of year-class strength derived from different sampling programs. However, we found no relationships between PCB exposure and any measure of striped bass abundance or reproduction. Although inconsistent with the expected effects of PCB exposures, trends in all measures of striped bass abundance and reproductive success were consistent with the expected effects of striped bass harvest restrictions that were imposed during the 1980s. Our results demonstrate a need for caution in inferring risks to populations in nature from effects observed in laboratory studies. PMID- 12564892 TI - Arsenic contamination of Bangladesh paddy field soils: implications for rice contribution to arsenic consumption. AB - Arsenic contaminated groundwater is used extensively in Bangladesh to irrigate the staple food of the region, paddy rice (Oryza sativa L.). To determine if this irrigation has led to a buildup of arsenic levels in paddy fields, and the consequences for arsenic exposure through rice ingestion, a survey of arsenic levels in paddy soils and rice grain was undertaken. Survey of paddy soils throughout Bangladesh showed that arsenic levels were elevated in zones where arsenic in groundwater used for irrigation was high, and where these tube-wells have been in operation for the longest period of time. Regression of soil arsenic levels with tube-well age was significant. Arsenic levels reached 46 microg g(-1) dry weight in the most affected zone, compared to levels below l0 microg g(-1) in areas with low levels of arsenic in the groundwater. Arsenic levels in rice grain from an area of Bangladesh with low levels of arsenic in groundwaters and in paddy soils showed that levels were typical of other regions of the world. Modeling determined, even these typical grain arsenic levels contributed considerably to arsenic ingestion when drinking water contained the elevated quantity of 0.1 mg L(-1). Arsenic levels in rice can be further elevated in rice growing on arsenic contaminated soils, potentially greatly increasing arsenic exposure of the Bangladesh population. Rice grain grown in the regions where arsenic is building up in the soil had high arsenic concentrations, with three rice grain samples having levels above 1.7 microg g(-1). PMID- 12564893 TI - Millennial-scale records of atmospheric mercury deposition obtained from ombrotrophic and minerotrophic peatlands in the Swiss Jura Mountains. AB - Peat cores from two bogs were used to reconstruct changes in net atmospheric mercury deposition in Switzerland for the past 2-3 millennia. The two records were compared to assess the reliability of peat cores as archives of atmospheric mercury deposition. Net mercury accumulation rates and Hg(ex), an indicator of significant anthropogenic mercury contamination, were calculated for both cores. Both records showed stable background values (0.5-1.9 and 1.0-3.0 microg of Hg m( 2) yr(-1)). In both profiles, mercury accumulation rates began to increase during the 12th century, and Hg(ex) appeared during the 14th century. The late 19th and early 20th centuries have been studied in detail. The profiles match well with the history of local and global mercury emissions. The magnitude of increase from the pre-anthropogenic to anthropogenic period was also very similar in both records. Although the two sites are botanically very similar and lie only 3.5 km apart, accumulation rates at TGE were generally higher than those at EGR. This indicates that, although such records can be used to determine the chronologies of and relative changes in rates of atmospheric mercury deposition, differences in rates of mercury accumulation rates between sites do not necessarily indicate differences in deposition rates of mercury from the atmosphere. PMID- 12564894 TI - Global accounting of PCBs in the continental shelf sediments. AB - The recycling longevity of hydrophobic organic contaminants (HOCs) within the global environment is set by their permanent removal through processes such as degradation and burial in geological reservoirs. More than 90% of the global sediment burial of organic carbon (OC) occurs on the continental shelves, representing < 10% of the earth's ocean area. The propensity of HOCs to associate with organic matter, and the proximity of most population centers and thus presumed source areas to coastal regions, led us to investigate shelf sediments as a depository of significance for global HOC budgets. Here, the global inventory and burial fluxes of polychlorinated biphenyls (PCBs) in continental shelf sediments were estimated on a congener-specific basis from a database of 4214 distinct continental shelf surface sediment samples. To account for near urban sampling bias, the locations of each datum relative to nearest population center were classified as Local (< 1 km), Regional (1-10 km), or Remote (> 10 km), according to a comprehensive vector map of the world (Digital Chart of the World) in a GIS environment. The global inventory of, for instance, PCB congener 153 was 1200 ton (95% confidence limit maximum: 2100 ton; and minimum 720 ton). The Remote sub-basin of the North Atlantic contains approximately half of the global shelf sediment inventory for most of the PCB congeners studied. The shelf sediment inventories of individual PCB congeners constitute significant fractions of their recently updated cumulative historical global emissions estimates. The estimated inventory in the shelf corresponds to about 10% of maximum emission estimates for lower-chlorinated congeners. However, for the more bioaccumulable, higher-chlorinated, congeners the shelf reservoirs appear to account for up to 80% of the estimated maximum cumulative global emissions. These shelf inventories represent 1-6% of the global industrial production of PCBs. The global burial fluxes were estimated to be on the order of 8-24 ton/yr each for the eight major congeners investigated, again, with the shelf constituting a more significant removal sink of the more chlorinated congeners. The permanent removal into deeper shelf sediments of PCB153 and PCB180 suggests that the global environmental mean residence times of these pollutants are on the order of 110 and 70 years, respectively. Hence, even after production and direct releases have been halted, we may expect to be exposed to such persistent pollutants for decades and centuries to come. PMID- 12564895 TI - Chorioallantoic membranes indicate avian exposure and biomarker responses to environmental contaminants: a laboratory study with White Leghorn chickens (Gallus domesticus). AB - PCB and endosulfan concentrations in the chorioallantoic membrane (CAM) of white leghorn chickens (Gallus domesticus) were evaluated as indicators of hepatic cytochrome P450 isozyme activity in hens and chicks as well as toxicant concentrations in eggs and hens. Sixteen hens were randomly divided into four groups of four and dosed with a mixture of PCB105 (2,3,3',4,4' pentachlorobiphenyl), PCB156(2,3,3',4,4',5-hexachlorobiphenyl), PCB189 (2,3,3',4,4',5,5'-heptachlorobiphenyl), and technical grade endosulfan (3:1 ratio of alpha and beta isomers) at three different dose groups. The first 10 fertile eggs laid by each hen were collected, the even-numbered eggs incubated until hatched, and the odd numbered eggs were analyzed for test chemicals. Strong (r2), significantly positive (p value) relationships were found between total PCB mass (ng) in CAMs and both total PCB concentrations (ng/g wet wt) in adults (r2 = 0.91, p = 0.0001) and eggs (r2 = 0.87, p = 0.0001). The relationship between total PCB mass in CAMs and hepatic cytochrome p450 isozyme activity in chicks (r2 = 0.49, p = 0.0001) and hens (r2 = 0.45, p = 0.014) was also significant but not as strong. This study shows that CAMs can be used to estimate avian exposure to PCBs and resultant biological response. PMID- 12564896 TI - Metal selectivity of Sargassum spp. and their alginates in relation to their alpha-L-guluronic acid content and conformation. AB - The discovery of a consistent and unusual enrichment in homopolymeric alpha-L guluronic acid G-blocks in alginates extracted from a suite of Sargassum brown algae is described in this study. 1H NMR spectroscopy was used to characterize these alginates which display homopolymeric guluronic acid block (G-block) frequency values (F(GG)) between 0.37 and 0.81. The presence of these G-blocks results in an enhanced selectivity for cadmium or calcium relative to monovalent ions such as sodium and the proton as well as smaller divalent ions such as magnesium. Results of competitive exchange experiments for the Cd-Ca-alginate system yield selectivity coefficient, K*(Cd)Ca, values between 0.43 +/- 0.10 and 1.32 +/- 0.02 for a range in F(GG) of 0.23 to 0.81. In contrast to the Cd-Ca alginate system, the Mg-Ca-alginate and Mg-Cd-alginate systems yielded maximum values of K*(Mg)Ca (18.0 +/- 1.4) and K*(Mg)Cd (16.0 +/- 0.9) for the alginates extracted from Sargassum fluitans (F(GG) = 0.81; Cuba) and Sargassum thunbergii (F(GG) = 0.75; Korea), respectively. Selectivity studies with mixed-metal pair alginate systems highlight the importance of the specific macromolecular conformation of the alginate polymer in determining metal binding behavior in multiple-metal systems. Furthermore, they demonstrate the importance of the conformation of the alginate as it occurs within the tissue of Sargassum in determining the metal binding behavior of this algal biosorbent. The unique composition of the alginates present in species of Sargassum may represent a distinct advantage over other brown algal species when considering their implementation for the strategic removal of toxic heavy metals from contaminated and industrial wastewaters. PMID- 12564897 TI - Measured pore-water concentrations make equilibrium partitioning work--a data analysis. AB - There is an increasing body of evidence that the bioaccumulation of sediment associated hydrophobic organic compounds (HOCs) is strongly influenced by sequestration. At present, it is not known how equilibrium partitioning theory (EqP), the most commonly employed approach for describing sediment bioaccumulation can be applied to sediments with sequestered contaminants. In this paper, we present freely dissolved pore-water concentrations of HOCs. These data were employed to interpret sediment bioaccumulation and sequestration data in order to arrive at a process based evaluation of EqP. The data analysis suggests that sediment bioaccumulation of compounds up to log K(ow) 7.5 in Tubificidae can be described as bioconcentration from pore-water. In addition, the pore-water concentrations of HOCs (4.5 < log K(ow) < 7.5) are established by equilibrium partitioning between the rapidly desorbing HOCs fraction in the sediment and the pore-water. Taken together, these findings indicate that EqP is a conceptually correct representation of sediment bioaccumulation, provided that sequestration is accounted for. This implies that the risk assessment of sediment associated HOCs can be significantly simplified: With a method at hand for measuring freely dissolved pore-water concentrations of HOCs, it appears that HOCs' body residues in sediment dwelling organisms can be estimated on the basis of concentrations in pore-water and bioconcentration factors. PMID- 12564898 TI - Impact of chlorine emissions from sea-salt aerosol on coastal urban ozone. AB - The ability of photochemical models to predict observed coastal chlorine levels and their corresponding effect on ozone formation is explored. Current sea-spray generation functions, a comprehensive gas-phase chlorine chemistry mechanism, and several heterogeneous/multiphase chemical reactions considered key processes leading to reactive chlorine formation are added to an airshed model of the South Coast Air Basin of California. Modeling results reproduce regional sea-salt particle concentrations. The heterogeneous/multiphase chemical reactions do not affect the rate of hydrochloric acid displacement, nor do they enhance aerosol nitrate formation. Chlorine levels in the model are predicted to be an order of magnitude lower than previously observed values at other coastal regions under similar conditions, albeit in much better agreement than previous studies. The results suggest that the inclusion of sea-salt-derived chlorine chemistry might increase morning ozone predictions by as much as 12 ppb in coastal regions and by 4 ppb in the peak domain ozone in the afternoon. The inclusion of anthropogenic sources of chlorine is recommended for future studies, as such sources might elevate ozone predictions even further via direct emission into polluted regions. PMID- 12564899 TI - Differential adsorption of molybdate and tetrathiomolybdate on pyrite (FeS2). AB - Molybdenum is a nutrient important for a variety of biological functions, most notably nitrogen fixation. Molybdenum availability is limited through sorption reactions, particularly in environments rich in sulfide minerals. This study examines the sorption of two major molybdenum species, molybdate (MoO4(2-)) and tetrathiomolybdate (MoS4(2-)), on synthetic pyrite (FeS2) as a function of solution composition. Both MoO4(2-) and MoS4(2-) partitioned strongly on FeS2 under a range of conditions and ionic strengths. Molybdate and tetrathiomolybdate adsorption obeyed a Langmuir isotherm with a calculated site density between 2 and 3 sites/nm2 under acidic and circumneutral conditions, which decreased to less than 1 site/ nm2 at pH 9. Although both MoO4(2-) and MoS4(2-) adsorbed most strongly under moderately acidic conditions, MoO4(2-) readily desorbed while MoS4(2-) remained adsorbed even at high pH. The reversibility of MoO4(2-) adsorption suggests the formation of labile surface complexes while MoS4(2-) likely forms strong inner-sphere complexes. X-ray absorption spectroscopy was used to determine the structure of the surface complexes. Molybdate formed bidentate, mononuclear complexes on FeS2. The Mo-S and Mo-Fe distances for tetrathiomolybdate on pyrite are consistent with the formation of Mo-Fe-S cubane type clusters. The high affinity of MoS4(2-) for FeS2, as well as its resistance to desorption, supports the hypothesis that thiomolybdate species are the reactive Mo constituents in reduced sediments and may control Mo enrichment in anoxic marine environments. PMID- 12564900 TI - Hydrodynamics of biological aggregates of different sludge ages: an insight into the mass transport mechanisms of bioaggregates. AB - Internal permeation and its role in mass transport to biological aggregates are investigated through detailed settling experiments. Three groups of bacterial aggregates (0.8-3.2 mm), which were different in fractal dimension and porosity, were generated in batch activated sludge reactors with biomass residence times (BRTs) of 5, 10, and 20 days. An apparatus of vertically connected double settling columns, which were filled respectively with water and an EDTA solution of a higher density, was used to characterize the settling and permeability features of individual aggregates. The settling velocities observed in water were in good agreement with those predicted by Stokes' law for porous but impermeable aggregates. The BRT10 and BRT20 aggregates, with porosities that were generally less than 0.92, had fluid collection efficiencies ranging from 0 to 0.1, while the BRT5 aggregates with porosities that were generally higher than 0.97 were almost completely impermeable. It is suggested that bioaggregates with either a tightly or a loosely packed structure cannot be as highly permeable as characterized for nonbiological fractal aggregates. While falling from water into the denser EDTA liquid, many BRT10 and BRT20 aggregates stopped and stayed below the interface of the two liquids for a period that was more than an order of magnitude shorter than the prediction based on the assumption of molecular diffusion as the predominant mass transport mechanism to the aggregates. The results indicate that limited intra-aggregate convection, which may not be important to the hydrodynamic properties of bioaggregates, could significantly enhance mass transport to suspended aggregates in biological wastewater treatment reactors. PMID- 12564901 TI - Sorption versus biomineralization of Pb(II) within Burkholderia cepacia biofilms. AB - X-ray spectroscopy measurements have been combined with macroscopic uptake data and transmission electron microscopy (TEM) results to show that Pb(II) uptake by Burkholderia cepacia is due to simultaneous sorption and biomineralization processes. X-ray microprobe mapping of B. cepacia biofilms formed on alpha-Al2O3 surfaces shows that Pb(II) is distributed heterogeneously throughout the biofilms because of the formation of Pb "hot spots". EXAFS data and TEM observations show that the enhanced Pb accumulation is due to the formation of nanoscale crystals of pyromorphite (Pb5(PO4)3(OH)) adjacent to the outer-membrane of a fraction of the total population of B. cepacia cells. In contrast, B. cepacia cell suspensions or biofilms that were heat-killed or pretreated with X-rays do not form pyromorphite, which suggests that metabolic activity is required. Precipitation of pyromorphite occurs over several orders of magnitude in [H-] and [Pb] and accounts for approximately 90% of the total Pb uptake below pH 4.5 but only 45-60% at near-neutral pH because of the formation of additional Pb(II) adsorption complexes. Structural fits of Pb L(III) EXAFS data collected for heat treated cells at near-neutral pH suggest that Pb(II) forms inner-sphere adsorption complexes with carboxyl functional groups in the biofilms. PMID- 12564902 TI - Rate constants for the gas-phase reactions of a series of alkylnaphthalenes with the nitrate radical. AB - Naphthalene and its methyl-, ethyl-, and dimethyl-derivatives are semivolatile polycyclic aromatic hydrocarbons expected to be in the gas phase in ambient atmospheres and are subject to nighttime degradation by gas-phase reactions with the nitrate (NO3) radical. Using a relative rate method, rate constants for the gas-phase reactions of NO3 radicals with a series of alkylnaphthalenes have been measured at 298 +/- 2 K and atmospheric pressure of air. The compounds studied were 1- and 2-methylnaphthalene (1- and 2-MN), 1- and 2-ethylnaphthalene (1- and 2-EN), and the 10 dimethylnaphthalene isomers (1,2-, 1,3-, 1,4-,1,5-, 1,6-, 1,7-, 1,8-, 2,3-, 2,6-, and 2,7-DMN). Sampling in Riverside, CA showed that these alkylnaphthalenes were readily detected in ambient air, with the exception of 1,8 DMN. The reactions of naphthalene and the alkylnaphthalenes with NO3 radicals proceed by initial addition of the radical to form an aromatic-NO3 adduct (with rate constant k(a)) which either decomposes back to reactants (with rate constant kb) or reacts with NO2 to form products (with rate constant k(c). Using naphthalene as the reference compound, the values of (k(a)k(c)/k(b)) obtained for the NO3 radical reactions (in units of 10(-28) cm(6) molecule(-2) S(-1), indicated errors are two least-squares standard deviations) were as follows: 1 MN, 7.15 +/- 0.37; 2-MN, 10.2 +/- 1.0; 1-EN, 9.82 +/- 0.69; 2-EN, 7.99 +/- 0.99; 1,2-DMN, 64.0 +/- 2.3; 1,3-DMN, 21.3 +/- 1.2; 1,4-DMN, 13.0 +/- 0.5; 1,5-DMN, 14.1 +/- 1.3; 1,6-DMN, 16.5 +/- 1.8; 1,7-DMN, 13.5 +/- 0.7; 1,8-DMN, 212 +/- 59; 2,3-DMN, 15.2 +/- 0.5; 2,6-DMN, 21.2 +/- 1.6; 2,7-DMN, 21.0 +/- 1.5. PMID- 12564903 TI - Bioaccumulation of LAS in feral fish studied by a novel LC-MS/MS method. AB - The extent of bioaccumulation of linear alkylbenzene sulfonate (LAS) in feral organisms is presently unknown. To enable LAS determination in biota samples, LAS and its coproducts (methylbranched LAS, dialkyltetralin sulfonates) are extracted from tissues using matrix solid-phase dispersion, isolated by strong anion exchange chromatography and determined by HPLC-electrospray-tandem mass spectrometry. All analytes were quantified in sediment dwelling Tubifex sp. with the sum of the concentrations exceeding 1 micromol/g. Since a portion of LAS was present adsorbed to solids in the guts, the actual body residue was lower than reported lethal LAS body residues. The concentrations of individual constituents in bream muscle samples from the river Saar and fathead minnows caged in the river Arrone ranged up to 2 nmol/g. The apparent bioaccumulation factors in the caged fish are consistently higher than laboratory data, presumably due to a combination of LAS present in the guts adsorbed to suspended material, coingestion of LAS with bacterial detritus, and depressed metabolic activity due to sublethal effects. Given the small deviation between laboratory and field bioaccumulation data and the high detection frequency (> 90%) of C(13)-2- and C13 iso-LAS, the latter two constituents are suitable markers for LAS contamination in fish. PMID- 12564904 TI - Fate of nonylphenol ethoxylates and their metabolites in two Dutch estuaries: evidence of biodegradation in the field. AB - The environmental behavior of nonylphenol ethoxylates (A9PEO) and their metabolites was investigated in field studies in the two Dutch estuaries Western Scheldt and the Rhine estuary. Using liquid chromatography-electrospray mass spectrometry (LC-ES-MS) analysis after solid-phase extraction, A9PEO, nonylphenol (NP), and the carboxylated metabolites (A9PEC) were determined in surface water and sediments. Maximum dissolved concentrations of 2.3, 0.9, and 8.1 microg L( 1), respectively, were found. In sediments, maximum concentrations of 242 and 1080 ng g(-1) for A9PEO and NP were observed. In almost half of the sediment samples, concentrations of A9PEC in sediments were below the detection limit. Occasionally relatively high values were observed, with a maximum of 239 ng g( 1). Metabolites of the carboxy alkylphenoxy ethoxy acetic acids (CAPEC) type could not be detected in any of the sediment or water samples. In the Scheldt estuary, dissolved concentration profiles showed nonconservative behaviorfor all detected compound groups. While A9PEO and NP concentrations strongly decreased along the salinity gradient, this decrease was weaker for the A9PEC metabolites. The increasing concentration ratio of A9PEC/A9PEO clearly illustratesthe important role that aerobic biodegradation plays in the estuarine fate of these compounds. It is concluded that the oxidative hydrolytic degradation pathway is the main degradation route in this nonstratified estuary. At high salinities, where concentrations drop to background levels of around 50 ng L(-1), this ratio decreases to about unity. Simple model calculations show that this can be explained if continuous diffuse discharges (e.g. from the intensive shipping in the estuary) are assumed. For the stratified Rhine estuary the water concentration profiles are less pronounced, possibly due to more complicated and turbulent water flows and point sources from the Rotterdam harbors. PMID- 12564905 TI - Characterization and copper binding of humic and nonhumic organic matter isolated from the South Platte River: evidence for the presence of nitrogenous binding site. AB - Humic substances typically constitute 40-60% of the dissolved organic matter (DOM) in surface waters. However, little information is available regarding the metal binding properties of the nonhumic hydrophilic portion of the DOM. In this study, humic and nonhumic DOM samples were isolated from the South Platte River (Colorado, DOC = 2.6 mg x L(-1), SUVA254 = 2.4 L/mg x m) using a two-column array of XAD-8 and XAD-4 resins. The three major isolated fractions of DOM, which accounted for 57% of the bulk DOM,were characterized using a variety of analytical tools. Proton and copper binding properties were studied for each fraction. The main objective of this work was to compare the structural and chemical characteristics of the isolated fractions and test models describing DOM reactivity toward metal ions. The characterization work showed significant structural differences between the three isolated fractions of DOM. The hydrophobic acid fraction (i.e., humic substances isolated from the XAD-8 resin) gave the largest C/H, C/O, and C/N ratios and aromatic carbon content among the three isolated fractions. The transphilic acid (TPHA) fraction ("transphilic" meaning fraction of intermediate polarity isolated from the XAD-4 resin) was found to incorporate the highest proportion of polysaccharides, whereas the transphilic neutral (TPHN) fraction was almost entirely proteinaceous. The gradual increase of the charge with pH for the three DOM fractions is most likely caused by a large distribution of proton affinity constants for the carboxylic groups, as well as a second type of group more generally considered to be phenolic. In the case of the DOM fraction enriched in proteinaceous material (i.e., TPHN fraction), the results showed that the amino groups are responsible for the charge reversal. For low copper concentrations, nitrogen-containing functional groups similar to those of amino acids are likely to be involved in complexation, in agreement with previously published data. PMID- 12564906 TI - The identification of plant derived structures in humic materials using three dimensional NMR spectroscopy. AB - Here we demonstrate the application of 3-D NMR spectroscopy to structural studies of humic substances, the most abundant of organic compounds on earth. The increased spectral dispersion provided by the additional dimension is proven to be highly advantageous in separating the overlapping signals observed in 2-D spectra. Assignments of the major aliphatic structures and selected aromatic moieties are given as examples. We find that in a forest soil fulvic acid the major aliphatic materials are likely derived from leaf cuticles and further demonstrate that lignin signatures can be identified among the aromatic species. Once identified from the 3-D spectra, these structures can be assigned using the partial information available in 2-D, and in some cases, in the 1-D spectra. These signals are demonstrated to be characteristic to given samples of natural organic matter, and the case is made for their use as indicators of terrestrial biomarkers in mixtures of compounds with unknown origins. PMID- 12564907 TI - Real-time PCR quantification of nitrifying bacteria in a municipal wastewater treatment plant. AB - Real-time PCR assays using TaqMan or Molecular Beacon probes were developed and optimized for the quantification of total bacteria, the nitrite-oxidizing bacteria Nitrospira, and Nitrosomonas oligotropha-like ammonia oxidizing bacteria (AOB) in mixed liquor suspended solids (MLSS) from a municipal wastewater treatment plant (WWTP) using a single-sludge nitrification process. The targets for the real-time PCR assays were the 16S rRNA genes (16S rDNA) for bacteria and Nitrospira spp. and the amoA gene for N. oligotropha. A previously reported assay for AOB 16S rDNA was also tested for its application to activated sludge. The Nitrospira 16S rDNA, AOB 16S rDNA, and N. oligotropha-like amoA assays were log linear over 6 orders of magnitude and the bacterial 16S rDNA real-time PCR assay was log-linear over 4 orders of magnitude with DNA standards. When these real time PCR assays were applied to DNA extracted from MLSS, dilution of the DNA extracts was necessary to prevent PCR inhibition. The optimal DNA dilution range was broad for the bacterial 16S rDNA (1000-fold) and Nitrospira 16S rDNA assays (2500-fold) but narrow for the AOB 16S rDNA assay (10-fold) and N. oligotropha like amoA real-time PCR assay (5-fold). In twelve MLSS samples collected over one year, mean cell per L values were 4.3 +/- 2.0 x 10(11) for bacteria, 3.7 +/- 3.2 x 10(10) for Nitrospira, 1.2 +/- 0.9 x 10(10) for all AOB, and 7.5 +/- 6.0 x 10(9) for N. oligotropha-like AOB. The percent of the nitrifying population was 1.7% N. oligotropha-like AOB based on the N. oligotropha amoA assay, 2.9% total AOB based on the AOB 16S rDNA assay, and 8.6% nitrite-oxidizing bacteria based on the Nitrospira 16S rDNA assay. Ammonia-oxidizing bacteria in the wastewater treatment plant were estimated to oxidize 7.7 +/- 6.8 fmol/hr/cell based on the AOB 16S rDNA assay and 12.4 +/- 7.3 fmol/hr/cell based on the N. oligotropha amoA assay. PMID- 12564908 TI - Using a passive multilayer sampler for measuring detailed profiles of gas-phase VOCs in the unsaturated zone. AB - The purpose of this study was to test in the laboratory the performance of a passive multilayer sampler (MLS) for obtaining detailed profiles of gas-phase volatile organic compounds (VOCs) in unsaturated sediments. The MLS is essentially a chain of isolated, cylindrical stainless steel dialysis cells filled with distilled water and closed with membranes at both ends. The sampling principle is based on passive equilibration of the unsaturated zone gas phase with water in the cells. Using trichloroethene (TCE) as a model VOC, and after testing the required equilibration time in the laboratory (about 50 h), results of a large container (210 L) experiment show that TCE concentrations obtained by the MLS deployed inside a well screen corresponded very well to the profile obtained by dialysis cells buried in the sediment. A field profile taken at the saturated-unsaturated interface region of a VOC-contaminated area using the MLS shows steep TCE concentration gradients (1119 microg TCE/L-air/cm) in the gas phase of the unsaturated zone just above the water table. PMID- 12564909 TI - Temperature-dependent uptake rates of nonpolar organic compounds by semipermeable membrane devices and low-density polyethylene membranes. AB - The effect of temperature on sampling rates and sampler-water partition coefficients of semipermeable membrane devices (SPMDs) and low-density polyethylene (LDPE) strips was studied in an experimental setup under controlled flow conditions. Aqueous concentrations of chlorobenzenes, polychlorinated biphenyls (PCBs), and polyaromatic hydrocarbons (PAHs) were maintained by continuous circulation of the water over a generator column. Sampling rates for standard design SPMDs (460 cm2) were in the range of 20-200 L d(-1). No significant differences were observed between sampling rates of SPMDs and LDPE strips, but the latter samplers reached equilibrium faster because of their smaller sorption capacity. Sampling rates at 30 degrees C were higher than at 2 degrees C by a factor of about 3. Sampling rate modeling indicated boundary layer controlled uptake for compounds with log octanol-water partition coefficients smaller than 4.4 and aqueous boundary-layer controlled uptake for more hydrophobic compounds. SPMD-water partition coefficients did not significantly change with temperature, but LDPE-water partition coefficients were larger at 2 degrees C than at 30 degrees C by a factor of 2. For field application of SPMDs, the results imply that temperature is not a key factor that controls uptake rates unless large geographical and temporal scales are involved. The results confirm that water flow velocity has a profound effect on sampling rates. PMID- 12564910 TI - Toxicity characteristic leaching procedure and iron treatment of brass foundry waste. AB - The Toxicity Characteristic Leaching Procedure (TCLP) is used by the United States Environmental Protection Agency to determine if wastes contain extractable components subject to hazardous waste regulations. This paper examines the limitations of the TCLP and the way it is used by studying a particular example. Waste casting sand from brass foundries to which iron metal has been added passes the TCLP test but when placed in a landfill for several years may start to leach lead, copper, and zinc. Results of TCLP tests of waste sand alone and with the additives iron metal, zinc metal, hydrous ferric oxide, and hematite are reported. Three processes were studied: reduction by metallic iron, sorption by hydrous ferric oxide, and precipitation of hydroxides. Lead, copper, and zinc behave differently with respect to these three processes, and their measurement allows some deductions as to what is occurring in a TCLP test or a landfill. Iron addition does not result in long-term stabilization of a waste placed in the ground. The chemistry of a laboratory extraction can be very different from the chemistry of a waste placed in the environment. Wastes that are treated to pass the TCLP test, but are not permanently stabilized, are a threat to the environment. PMID- 12564911 TI - Electron-beam treatment of aromatic hydrocarbons that can be air-stripped from contaminated groundwater. 1. Model studies in aqueous solution. AB - As a model for the electron-beam degradation of volatile aromatics (benzene, toluene, ethylbenzene, xylenes, BTEX) in groundwater strip gas, to be reported in Part 2, the gamma-radiolysis of benzene has been studied in aqueous solutions. Addition of *OH to the aromatic ring gives rise to hydroxycyclohexadienyl radicals which either dimerize or disproportionate. The various dimers undergo acid-catalyzed water elimination yielding biphenyl. Phenol is formed upon disproportionation directly, but also via dihydroxycyclohexadiene which subsequently undergoes acid-catalyzed water elimination. Co-radiolysis of benzene with nitrite generates *NO2 in addition to the hydroxycyclohexadienyl radical. These not only interact with one another (product: nitrobenzene via nitro hydroxycyclohexadienes) but the *NO2 radical is also capable of abstracting cyclohexadienylic hydrogens. This reaction leads to the formation of 2- and 4 nitrophenol and to further nitrated products that were not identified. These are suggested to be formed in an analogous reaction of *NO2 with the hydroxycylohexadienyl dimers. The effect of O2 on these reactions and the relevance for the gas-phase radiolysis of BTEX is discussed. PMID- 12564912 TI - Electron-beam treatment of aromatic hydrocarbons that can be air-stripped from contaminated groundwater. 2. Gas-phase studies. AB - The electron-beam (EB) degradation of volatile aromatics (benzene, toluene, ethylbenzene, xylenes: BTEX) in groundwater strip gas, which in the present work has been modeled by the introduction of the desired aromatic(s) to a stream of air or another gas, such as oxygen, is initiated essentially by the addition of *OH radicals to the aromatic ring, giving rise to hydroxycyclohexadienyl radicals, which form the corresponding peroxyl radicals upon addition of oxygen. As studied in some detail with benzene as a BTEX representative, various reactions of these lead to numerous oxidation products in a cascade of reactions, including the decomposition of products under the prevailing conditions of high turnover of the initial aromatic. Importantly, hydroxycyclohexadienylperoxyl radical formation is partly reversible, and the reactions of the hydroxycyclohexadienyl radicals, which thus have a significant presence in these systems, must therefore also be taken into consideration. In the gas phase, in contrast to the aqueous phase (see Part 1), the reactions of the hydroxycyclohexadienyl radicals lead to oligomeric products that appear to contribute, in addition to ionic clusters, to nucleation for the aerosols observed. Various nitrated products, among them nitrophenols, are observed when air is used for the stripping. However, these studies did not clear the pilot plant stage, since BTEX degradation using a bioreactor carried out in parallel was so successful that the EB technology was judged to be noncompetitive. As for the latter, expensive equipment consisting of a stripper, the EB machine, and an aerosol precipitator would be required. The condensed aerosols are biorefractory and would require further treatment for detoxification. PMID- 12564913 TI - Effects of humic substances on the pattern of oxidation products of pentachlorophenol induced by a biomimetic catalytic system using tetra(p sulfophenyl)porphineiron(III) and KHSO5. AB - In the presence of humic substances (HSs), the oxidative conversion of pentachlorophenol (PCP) was found to be efficiently catalyzed by tetra(p sulfophenyl)porphineiron(III) (Fe(III)-TPPS) using KHSO5 as an oxygen donor. Ortho-tetrachloroquinone (o-TeCQ), 2-hydroxyl-nonachlorodiphenyl ether (2H-NCDE), 4-hydroxyl-nonachlorodiphenyl ether (4H-NCDE), and octachlorodibenzo-p-dioxin (OCDD) were identified as the major byproducts of the reaction. Decreased amounts of these byproducts were produced in the presence of HS. In particular, the addition of HSs with a lower degree of humification resulted in a large decrease in the formation of dimers, such as 2H-NCDE, 4H-NCDE, and OCDD. More than 60% of the chlorine, which was released from PCP, was found in the HS fractions after the reaction. This suggests that chlorinated intermediates from PCP were incorporated into the HS. Pyrolysis-GC/MS and 13C NMR studies confirmed that the binding of the chlorinated intermediates was covalent in nature and that the intermediates were copolymerized with HS via oxidative coupling reactions. A Microtox test demonstrated that the toxicity of the HS fraction containing PCP derived intermediates was much lower than that of the mixture of PCP and HS in the absence of a catalytic reaction. PMID- 12564914 TI - Remote sensing study of emissions from motor vehicles in the Metropolitan Area of Mexico City. AB - Remote sensing was employed for the first time to measure nitric oxide (NO) levels of on-road light-duty motor vehicles of the Metropolitan Area of Mexico City (MAMC). The sensor placed at 12 different sites also measured the concentration of CO2, CO, and total hydrocarbons (THC) in the exhaust emissions. A database was compiled containing 122 800 readings, of which 84 650 (69%) records were valid emissions measurements. CO, HC, and NO valid readings were 68.9, 63.4, and 62.9%, respectively, of the total attempted measurements. Furthermore, 42 822 vehicles were number-plate-matched to model year with the information provided by the Inspection/Maintenance Program. The mean emissions of total valid readings for CO, HC, and NO were determined to be 1.31 vol %, 440 ppm (propane), and 914 ppm, respectively. In 1991 and 1994, remote sensing measurements of CO and HC tailpipe emissions were performed in the MAMC in five different locations (30 000 valid readings). Large drops in both pollutants were observed for the intervening years, but sufficient vehicle information was not available at that time to fully explain the observed trends. Compared with those reports, our results point out to a steady decrease in CO and HC exhaust emissions with vehicle model year. The fleet emissions measured exhibit a gamma distribution, with 10% of the most polluting fleet studied being responsible for 45%, 25%, and 29% of the CO, HC, and NO emissions, respectively. NO emissions in taxis are the highest among the vintage of vehicles, a matter of concern since according to the distance traveled per year, they represent 22% of the total activity in the MAMC. PMID- 12564915 TI - Activated carbon as an electron acceptor and redox mediator during the anaerobic biotransformation of azo dyes. AB - Activated carbon (AC) has a long history of applications in environmental technology as an adsorbent of pollutants for the purification of drinking waters and wastewaters. Here we describe novel role of AC as redox mediator in accelerating the reductive transformation of pollutants as well as a terminal electron acceptor in the biological oxidation of an organic substrate. This study explores the use of AC as an immobilized redox mediator for the reduction of a recalcitrant azo dye (hydrolyzed Reactive Red 2) in laboratory-scale anaerobic bioreactors, using volatile fatty acids as electron donor. The incorporation of AC in the sludge bed greatly improved dye removal and formation of aniline, a dye reduction product. These results indicate that AC acts as a redox mediator. In supporting batch experiments, bacteria were shown to oxidize acetate at the expense of reducing AC. Furthermore, AC greatly accelerated the chemical reduction of an azo dye by sulfide. The results taken as a whole clearly suggest that AC accepts electrons from the microbial oxidation of organic acids and transfers the electrons to azo dyes, accelerating their reduction. A possible role of quinone surface groups in the catalysis is discussed. PMID- 12564916 TI - Sorption hysteresis of benzene in charcoal particles. AB - Charcoal is found in water, soil, and sediment where it may act as a sorbent of organic pollutants. The sorption of organic compounds to natural solids often shows hysteresis. The purpose of this study was to determine the source of pronounced hysteresis that we found in the sorption of a hydrophobic compound (benzene) in water to a maple-wood charcoal prepared by oxygen-limited pyrolysis at 673 K. Gas adsorption (N2, Ar, CO2), 13C NMR, and FTIR show the charcoal to be a microporous solid composed primarily of elemental (aromatic) C and secondarily of carboxyl and phenolic C. Nonlocal density functional theory (N2, Ar) and Monte Carlo (CO2) calculations reveal a porosity of 0.15 cm3/g, specific surface area of 400 m2/g, and appreciable porosity in ultramicropores < 10 A. Benzene sorption desorption conditions were chosen to eliminate artificial causes of hysteresis (rate-limiting diffusion, degradation, colloids effect). Charcoal sorbed up to its own weight of benzene at approximately 69% of benzene water solubility. Sorption was highly irreversible over most of the range tested (10(-4)-10(3) microg/mL). A dimensionless irreversibility index (/i) (0 < or = /i < or = 1) based on local slopes of adsorption and desorption branches was evaluated at numerous places along the isotherm. /i decreases as C increases, from 0.9-1 at low concentration to approximately 0 (approximately fully reversible) at the highest concentrations. Using sedimentation and volumetric displacement measurements, benzene is observed to cause pronounced swelling (up to > 2-fold) of the charcoal particles. It is proposed that hysteresis is due to pore deformation by the solute, which results in the pathway of sorption being different than the pathway of desorption and which leads to entrapment of some adsorbate as the polyaromatic scaffold collapses during desorption. It is suggested that intra-charcoal mass transport may be influenced by structural rearrangement of the solid, in addition to molecular diffusion. PMID- 12564917 TI - Decomposition of environmentally persistent trifluoroacetic acid to fluoride ions by a homogeneous photocatalyst in water. AB - Decomposition of trifluoroacetic acid (TFA) was achieved with a tungstic heteropolyacid photocatalyst H3PW12O40*6H2O in order to develop a technique for measures against TFA stationary sources. This is the first example of C-F bond cleavage in an environmentally harmful perfluoromethyl-group-containing compound using a homogeneous photocatalyst. The catalytic reaction proceeds in water at room temperature under UV-visible light irradiation in the presence of oxygen. The system produces only F- ions and CO2; the (mole of formed F-)/(mole of decomposed TFA) and (mole of formed CO2)/(mole of decomposed TFA) ratios were 2.91 and 2.09, respectively. GC/MS measurements showed no trace of other species such as environmentally undesirable CF4, which is the most stable perfluorocarbon and has a very high global warming potential. When the (initial TFA)/(initial catalyst) molar ratio was 20:1, the turnover number of TFA decomposition reached 5.58 by 72 h of irradiation, accompanying with no catalyst degradation. The catalytic reaction mechanism can be explained by a redox reaction between the catalyst and TFA, involving a photo-Kolbe process. PMID- 12564918 TI - Reactive phosphorus removal from aquaculture and poultry productions systems using polymeric hydrogels. AB - This work reports on the features of a sorption processes for the ultimate removal and recovery of reactive phosphorus from aquaculture and poultry production wastewater effluents. The sorbent used was a cross-linked polyamine (PAA-HCl) polymeric hydrogel. The PAA-HCl hydrogels were prepared by chemically cross-linking aqueous solutions of linear PAA-HCl chains with epichlorohydrin (EPI). The phosphorus binding capacity of the gels was measured in standard aqueous solutions as a function of ionic strength. Equilibrium PO4(3-), loadings of 100 mg anion/g gel were obtained. The regeneration ability of the gels was demonstrated by release of the bound phosphorus anions upon washing with 1-2 M NaOH solution, providing opportunities to recover and reuse the gel over multiple cycles. The ionic polyamine gels have been demonstrated to be appropriate materials for treating poultry and aquaculture wastewater effluents. Upon treatment phosphorus anion concentrations were reduced to levels suitable for discharge into natural surface waters. PMID- 12564919 TI - Application of an isothermal, three-phase catalytic reactor model to predict unsteady-state fixed-bed performance. AB - CatReac, a three-phase catalytic mathematical model, was developed for analysis and optimization of the volatile reactor assembly used in International Space Station water processor. This wet oxidation process is used to remove low molecular weight contaminants such as acetic acid, acetone, ethanol, 1-propanol, 2-propanol, and propionic acid, which are not removed by the other treatment processes. The Langmuir-Hinshelwood (Hinshelwood, C. N. The Kinetics of Chemical Change in Gaseous Systems, 3rd ed.; Oxford: London, 1933; pp 301-347) isothermal adsorption expression was successfully used to describe the reaction kinetics of compounds on the catalyst surface for the compounds mentioned above. Small-column experiments combined with the use of the Arrhenius equation were successfully used to predict the Langmuir-Hinshelwood parameters under different temperatures for a temperature range from 93 to 149 degrees C. Full-scale and small-column experiments were successfully used to validate the model predictions for unsteady state fixed-bed operations. PMID- 12564920 TI - Disulfide bond isomerization in prokaryotes. PMID- 12564921 TI - New complex of post-activated neocarzinostatin chromophore with DNA: bulge DNA binding from the minor groove. AB - Neocarzinostatin (NCS-chrom), a natural enediyne antitumor antibiotic, undergoes either thiol-dependent or thiol-independent activation, resulting in distinctly different DNA cleavage patterns. Structures of two different post-activated NCS chrom complexes with DNA have been reported, revealing strikingly different binding modes that can be directly related to the specificity of DNA chain cleavage caused by NCS-chrom. The third structure described herein is based on recent studies demonstrating that glutathione (GSH) activated NCS-chrom efficiently cleaves DNA at specific single-base sites in sequences containing a putative single-base bulge. In this structure, the GSH post-activated NCS-chrom (NCSi-glu) binds to a decamer DNA, d(GCCAGAGAGC), from the minor groove. This binding triggers a conformational switch in DNA from a loose duplex in the free form to a single-strand, tightly folded hairpin containing a bulge adenosine embedded between a three base pair stem. The naphthoate aromatic moiety of NCSi glu intercalates into a GG step flanked by the bulge site, and its substituent groups, the 2-N-methylfucosamine carbohydrate ring and the tetrahydroindacene, form a complementary minor groove binding surface, mostly interacting with the GCC strand in the duplex stem of DNA. The bulge site is stabilized by the interactions involving NCSi-glu naphthoate and GSH tripeptide. The positioning of NCSi-glu is such that only single-chain cleavage via hydrogen abstraction at the 5'-position of the third base C (which is opposite to the putative bulge base) in GCC is possible, explaining the observed single-base cleavage specificity. The reported structure of the NCSi-glu-bulge DNA complex reveals a third binding mode of the antibiotic and represents a new family of minor groove bulge DNA recognition structures. We predict analogue structures of NCSi-R (R = glu or other substituent groups) may be versatile probes for detecting the existence of various structures of nucleic acids. The NMR structure of this complex, in combination with the previously reported NCSi-gb-bulge DNA complex, offers models for specific recognition of DNA bulges of various sizes through binding to either the minor or the major groove and for single-chain cleavage of bulge DNA sequences. PMID- 12564922 TI - The crystal structure of phosphonate-inhibited D-Ala-D-Ala peptidase reveals an analogue of a tetrahedral transition state. AB - D-Alanyl-D-alanine carboxypeptidase/transpeptidases (DD-peptidases) are beta lactam-sensitive enzymes that are responsible for the final peptidoglycan cross linking step in bacterial cell wall biosynthesis. A highly specific tripeptide phosphonate inhibitor was designed with a side chain corresponding to a portion of the Streptomyces R61 peptidoglycan. This compound was found to be a slow, irreversible inactivator of the DD-peptidase. Molecular modeling suggested that although a pentacoordinated intermediate of the phosphonylation reaction would not interact strongly with the enzyme, a tetracoordinated phosphonyl enzyme might be analogous to a transition state in the reaction with peptide substrates. To investigate this possibility, the crystal structure of the phosphonyl enzyme was determined. The 1.1 A resolution structure shows that the inhibitor has phosphonylated the catalytic serine (Ser62). One of the phosphonyl oxygens is noncovalently bound in the oxyanion hole, while the other is solvated by two water molecules. The conserved hydroxyl group of Tyr159 forms a strong hydrogen bond with the latter oxygen atom (2.77 A). This arrangement is interpreted as being analogous to the transition state for the formation of the tetrahedral intermediate in the deacylation step of the carboxypeptidase reaction. The proximity of Tyr159 to the solvated phosphonyl oxygen suggests that the tyrosine anion acts as a general base for deacylation. This transition state analogue structure is compared to the structures of noncovalent DD-peptidase reaction intermediates and phosphonylated beta-lactamases. These comparisons show that specific substrate binding to the peptidase induces a conformational change in the active site that places Ser62 in an optimal position for catalysis. This activated conformation relaxes as the reaction proceeds. PMID- 12564923 TI - Low-temperature-induced structural changes in human lysozyme elucidated by three dimensional NMR spectroscopy. AB - The three-dimensional solution structures of human lysozyme were determined at 35 and 4 degrees C using the heteronuclear multidimensional NMR spectroscopy, which were compared with each other to clarify the structural response of this enzyme to lowering of the temperature. Together with the data of the temperature dependence experiments of the lytic activity against Micrococcus luteus, we consider the implication of the observed structural change for the low temperature-induced reduction of the activity of human lysozyme. The structures of human lysozyme determined at the two temperatures are found to be similar, both of which comprise four alpha-helices (A- to D-helices) and three antiparallel beta-strands (beta(1)-beta(3)), leading to the constructions of the alpha- and beta-domains as previously identified in the X-ray crystal structure. A significant structural change was observed for the "active site lobe" comprising the loop region connecting C- and D-helices and the following D-helix, which moves toward the active site cleft located between the alpha- and beta domains so as to obstruct the cleft according to the temperature lowering. It further appeared that the total volume as well as the accessible surface area of human lysozyme decreases with lowering of the temperature, suggesting that the internal cavity of this enzyme shrinks under low temperature environment. Because in human lysozyme the region comprising the active site lobe is responsible for turnover of the enzymatic reaction against the substrate, the low-temperature induced structural change of the active site lobe presumably controls the efficiency of the lytic activity under low temperatures. PMID- 12564924 TI - Glutamine 132 in the NAD(H)-binding component of proton-translocating transhydrogenase tethers the nucleotides before hydride transfer. AB - Transhydrogenase, found in bacterial membranes and inner mitochondrial membranes of animal cells, couples the redox reaction between NAD(H) and NADP(H) to proton translocation. In this work, the invariant Gln132 in the NAD(H)-binding component (dI) of the Rhodospirillum rubrum transhydrogenase was substituted with Asn (to give dI.Q132N). Mixtures of the mutant protein and the NADP(H)-binding component (dIII) of the enzyme readily produced an asymmetric complex, (dI.Q132N)(2)dIII(1). The X-ray structure of the complex revealed specific changes in the interaction between bound nicotinamide nucleotides and the protein at the hydride transfer site. The first-order rate constant of the redox reaction between nucleotides bound to (dI.Q132N)(2)dIII(1) was <1% of that for the wild type complex, and the deuterium isotope effect was significantly decreased. The nucleotide binding properties of the dI component in the complex were asymmetrically affected by the Gln-to-Asn mutation. In intact, membrane-bound transhydrogenase, the substitution completely abolished all catalytic activity. The results suggest that Gln132 in the wild-type enzyme behaves as a "tether" or a "tie" in the mutual positioning of the (dihydro)nicotinamide rings of NAD(H) and NADP(H) for hydride transfer during the conformational changes that are coupled to the translocation of protons across the membrane. This ensures that hydride transfer is properly gated and does not take place in the absence of proton translocation. PMID- 12564925 TI - Palmitoylation of phospholipid scramblase 1 controls its distribution between nucleus and plasma membrane. AB - Phospholipid scramblase 1 (PLSCR1) is a Ca(2+)-binding, endofacial plasma membrane protein thought to contribute to the transbilayer movement of phosphatidylserine and other membrane phospholipids that is observed upon influx of calcium into the cytosol. Expression of PLSCR1 is markedly induced by interferon and other cytokines, and PLSCR1-/- bone marrow cells exhibit defective myeloid proliferation and differentiation in response to stimulation by select growth factors, implying that PLSCR1 also functions in cytokine signaling or response pathways. PLSCR1 is multiply palmitoylated and partitions into membrane lipid raft domains. We have now identified the Cys-rich sequence (184)CCCPCC(189) in PLSCR1 as required for palmitoylation of the polypeptide. Mutation of these five cysteines abrogates PLSCR1 trafficking to the plasma membrane and results in virtually all of the expressed protein localizing to the nucleus. Consistent with this observation, cell treatment with the palmitoylation inhibitor, 2-bromo palmitate, results in a marked redistribution of endogenous PLSCR1 from plasma membrane to nucleus. In a small percentage of untreated cells, predominantly nuclear localization of PLSCR1 is also observed. Furthermore, PLSCR1 is also found in the nucleus following its cytokine-induced expression. These data suggest that under the circumstance of rapid biosynthesis in response to gene induction by cytokines, PLSCR1 traffics into the nucleus, implying a potential nuclear function for this protein. PMID- 12564926 TI - Recognition of DNA interstrand cross-link of antitumor cisplatin by HMGB1 protein. AB - Several proteins that specifically bind to DNA modified by cisplatin, including those containing HMG-domains, mediate antitumor activity of this drug. Oligodeoxyribonucleotide duplexes containing a single, site-specific interstrand cross-link of cisplatin were probed for recognition by the rat chromosomal protein HMGB1 and its domains A and B using the electrophoretic mobility-shift assay. It has been found that the full-length HMGB1 protein and its domain B to which the lysine-rich region (seven amino acid residues) of the A/B linker is attached at the N-terminus (the domain HMGB1b7) specifically recognize DNA interstrand cross-linked by cisplatin. The affinity of these proteins to the interstrand cross-link of cisplatin is not very different from that to the major 1,2-GG intrastrand cross-link of this drug. In contrast, no recognition of the interstrand cross-link by the domain B lacking this region or by the domain A with or without this lysine-rich region attached to its C-terminus is noticed under conditions when these proteins readily bind to 1,2-GG intrastrand adduct. A structural model for the complex formed between the interstrand cross-linked DNA and the domain HMGB1b7 was constructed and refined using molecular mechanics and molecular dynamics techniques. The calculated accessible areas around the deoxyribose protons correlate well with the experimental hydroxyl radical footprint. The model suggests that the only major adaptation necessary for obtaining excellent surface complementarity is extra DNA unwinding (approximately 40 degrees ) at the site of the cross-link. The model structure is consistent with the hypothesis that the enhancement of binding affinity afforded by the basic lysine-rich A/B linker is a consequence of its tight binding to the sugar phosphate backbone of both DNA strands. PMID- 12564927 TI - Role of glutamic acid 216 in cytochrome P450 2D6 substrate binding and catalysis. AB - Human cytochrome P450 (P450) 2D6 is an important enzyme involved in the metabolism of drugs, many of which are amines or contain other basic nitrogen atoms. Asp301 has generally been considered to be involved in electrostatic docking with the basic substrates, on the basis of previous modeling studies and site-directed mutagenesis. Substitution of Glu216 with a residue other than Asp strongly attenuated the binding of quinidine, bufuralol, and several other P450 2D6 ligands. Catalytic activity with the substrates bufuralol and 4 methoxyphenethylamine was strongly inhibited by neutral or basic mutations at Glu216 (>95%), to the same extent as the substitution of Asn at Asp301. Unlike the Asp301 mutants, the Gln216 mutant (E216Q) retained 40% enzyme efficiency with the substrate spirosulfonamide, devoid of basic nitrogen, suggesting that the substitutions at Glu216 affect binding of amine substrates more than other catalytic steps. Attempts to induce catalytic specificity toward new substrates by substitutions at Asp301 and Glu216 were unsuccessful. Collectively, the results provide evidence for electrostatic interaction of amine substrates with Glu216, and we propose that both of these acidic residues plus at least another residue(s) is (are) involved in binding the repertoire of P450 2D6 ligands. PMID- 12564928 TI - C2 domain of protein kinase C alpha: elucidation of the membrane docking surface by site-directed fluorescence and spin labeling. AB - The C2 domain is a conserved signaling motif that triggers membrane docking in a Ca(2+)-dependent manner, but the membrane docking surfaces of many C2 domains have not yet been identified. Two extreme models can be proposed for the docking of the protein kinase C alpha (PKC alpha) C2 domain to membranes. In the parallel model, the membrane-docking surface includes the Ca(2+) binding loops and an anion binding site on beta-strands 3-4, such that the beta-strands are oriented parallel to the membrane. In the perpendicular model, the docking surface is localized to the Ca(2+) binding loops and the beta-strands are oriented perpendicular to the membrane surface. The present study utilizes site-directed fluorescence and spin-labeling to map out the membrane docking surface of the PKC alpha C2 domain. Single cysteine residues were engineered into 18 locations scattered over all regions of the protein surface, and were used as attachment sites for spectroscopic probes. The environmentally sensitive fluorescein probe identified positions where Ca(2+) activation or membrane docking trigger measurable fluorescence changes. Ca(2+) binding was found to initiate a global conformational change, while membrane docking triggered the largest fluorescein environmental changes at labeling positions on the three Ca(2+) binding loops (CBL), thereby localizing these loops to the membrane docking surface. Complementary EPR power saturation measurements were carried out using a nitroxide spin probe to determine a membrane depth parameter, Phi, for each spin labeled mutant. Positive membrane depth parameters indicative of membrane insertion were found for three positions, all located on the Ca(2+) binding loops: N189 on CBL 1, and both R249 and R252 on CBL 3. In addition, EPR power saturation revealed that five positions near the anion binding site are partially protected from collisions with an aqueous paramagnetic probe, indicating that the anion binding site lies at or near the surface of the headgroup layer. Together, the fluorescence and EPR results indicate that the Ca(2+) first and third Ca(2+) binding loops insert directly into the lipid headgroup region of the membrane, and that the anion binding site on beta-strands 3-4 lies near the headgroups. The data support a model in which the beta-strands are tilted toward the parallel orientation relative to the membrane surface. PMID- 12564929 TI - NMR structure of the DNA-binding domain of the cell cycle protein Mbp1 from Saccharomyces cerevisiae. AB - The three-dimensional solution structure of the DNA-binding domain of Mlu-1 box binding protein (Mbp1) has been determined by multidimensional NMR spectroscopy. Mbp1 is a cell cycle transcription factor from Saccharomyces cerevisiae and consists of an N-terminal DNA-binding domain, a series of ankyrin repeats, and a heterodimerization domain at the C-terminus. A set of conformers comprising 19 refined structures was calculated via a molecular dynamics simulated annealing protocol using distance, dihedral angle, and residual dipolar coupling restraints derived from either double or triple resonance NMR experiments. The solution structure consists of a six-stranded beta-sheet segment folded against two pairs of alpha-helices in the topology of the winged helix-turn-helix family of proteins and is in agreement with the X-ray structures. In addition, the solution structure shows that the C-terminal tail region of this domain folds back and makes specific interactions with the N-terminal beta-strand of the protein. This C-terminal region is essential for full DNA-binding activity but appears in the X ray structure to be disordered. The fold-back structure extends the region of positive electrostatic potential, and this may enhance the nonspecific contribution to binding by favorable electrostatic interactions with the DNA backbone. PMID- 12564930 TI - Mapping the microtubule binding regions of calponin. AB - The smooth muscle basic calponin interacts with F-actin and inhibits the actomyosin ATPase in a calmodulin or phosphorylation modulated manner. It also binds in vitro to microtubules and its acidic isoform, present in nonmuscle cells, and co-localizes with microfilaments and microtubules in cultured neurons. To assess the physiological significance and the molecular basis of the calponin microtubule interaction, we have first studied the solution binding of recombinant acidic calponin to microtubules using quantitative cosedimentation analyses. We have also characterized, for the first time, the ability of both calponin isoforms to induce the inhibition of the microtubule-stimulated ATPase activity of the cytoskeletal, kinesin-related nonclaret dysjunctional motor protein (ncd) and the abolition of this effect by calcium calmodulin. This property makes calponin a potent inhibitor of all filament-activated motor ATPases and, therefore, a potential regulatory factor of many motor-based biological events. By combining the enzymatic measurements of the ncd microtubules system with various in vitro binding assays employing proteolytic, recombinant and synthetic fragments of basic calponin, we further unambiguously identified the interaction of microtubules at two distinct calponin sites. One is inhibitory and resides in the segment 145-182, which also binds F-actin and calmodulin. The other one is noninhibitory, specific for microtubules, and is located on the COOH-terminal repeat-containing region 183-292. Finally, quantitative fluorescence studies of the binding of basic calponin to the skeletal pyrenyl F-actin in the presence of microtubules did not reveal a noticeable competition between the two sets of filaments for calponin. This result implies that calponin undergoes a concomitant binding to both F-actin and microtubules by interaction at the former site with actin and at the second site with microtubules. Thus, in the living cells, calponin could potentially behave as a cross-linking protein between the two major cytoskeletal filaments. PMID- 12564932 TI - Stability of drug-induced tubulin rings by fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) was applied to investigate the stability of tubulin rings that result from the interaction of alpha beta-tubulin dimers with three vinca domain-binding peptides--cryptophycin 1, hemiasterlin, and dolastatin 10. These peptides inhibit tubulin polymerization into microtubules and, instead, induce the formation of single-walled tubulin rings of 23.8 nm mean diameter for cryptophycin and 44.6 nm mean diameter for hemiasterlin and dolastatin, as revealed by electron microscopy on micromolar drug-tubulin samples. However, the hydrodynamic diameter and the apparent number of fluorescent particles, determined from analysis of FCS measurements obtained from nanomolar drug-tubulin samples, indicate variation in the stability of the rings depending on the drug and the tubulin concentration. Cryptophycin-tubulin rings appear to be the most stable even with tubulin concentration as low as 1 nM, whereas hemiasterlin-tubulin rings are the least, depolymerizing even at relatively high concentrations (100 nM). In contrast, the dolastatin-tubulin rings demonstrate an intermediate level of stability, depolymerizing significantly only at tubulin concentrations below 10 nM. We also compare the stability results with those of cytotoxicity measurements taken on several cell lines and note a rough correlation between the cytotoxicity of the drugs in cell cultures and the stability of the corresponding drug-induced rings. PMID- 12564931 TI - Hepatocyte apoB-containing lipoprotein secretion is decreased by the grapefruit flavonoid, naringenin, via inhibition of MTP-mediated microsomal triglyceride accumulation. AB - Naringenin, the principal flavonoid in grapefruit, reduces plasma lipids in vivo and inhibits apoB secretion, cholesterol esterification, and MTP activity in HepG2 human hepatoma cells. Although naringenin inhibits ACAT, we recently demonstrated that CE availability in the microsomal lumen does not regulate apoB secretion in HepG2 cells. We therefore hypothesized that inhibition of TG accumulation in the ER lumen, secondary to MTP inhibition, is the primary mechanism whereby naringenin blocks lipidation and subsequent secretion of apoB. Multicompartmental modeling of pulse-chase studies was used to compare cellular apoB kinetics in the presence of either naringenin or the specific MTP inhibitor, BMS-197636. At concentrations that reduced apoB secretion by 50%, both compounds selectively enhanced degradation via a kinetically defined, rapid, proteasomal pathway to the same extent. Subcellular fractionation experiments revealed that naringenin and BMS-197636 reduced accumulation of newly synthesized TG in the microsomal lumen by 48% and 54%, respectively. Newly synthesized CE accumulation in the lumen was reduced by 80% and 33% with naringenin and BMS-197636, respectively, demonstrating for the first time that MTP is involved in CE accumulation in the microsomal lumen. Reduced TG availability at this initial site of lipoprotein assembly was associated with significant reductions in the secretion of apoB-containing lipoproteins. Both naringenin and BMS-197636 were most effective in reducing secretion of IDL and LDL, but also inhibited secretion of apoB-containing HDL-sized particles. Furthermore, in McA-RH7777-derived cell lines, naringenin reduced secretion of hapoB72 and hapoB100, which require significant assembly with lipid to be secreted, but did not reduce secretion of hapoB17, hapoB23, and hapoB48, which require only minimal lipidation. Taken together, our results indicate that naringenin inhibits the lipidation and subsequent secretion of apoB-containing lipoproteins primarily by limiting the accumulation of TG in the ER lumen, secondary to MTP inhibition. PMID- 12564933 TI - Basal and hydrogen peroxide stimulated sites of phosphorylation in heterogeneous nuclear ribonucleoprotein C1/C2. AB - Hydrogen peroxide (H2O2) is a recently recognized second messenger, which regulates mammalian cell proliferation and migration. The biochemical mechanisms by which mammalian cells sense and respond to low concentrations of H2O2 are poorly understood. Recently, heterogeneous nuclear ribonucleoprotein C1/C2 (hnRNP C1/C2) was found to be rapidly phosphorylated in response to the application of low concentrations of H2O2 to human endothelial cells. Here, using tandem mass spectrometry, four sites of phosphorylation are identified in hnRNP-C1/C2, all of which are in the acidic C-terminal domain of the protein. Under resting conditions, the protein is phosphorylated at S247 and S286. In response to low concentrations of H2O2, there is increased phosphorylation at S240 and at one of the four contiguous serine residues from S225-S228. Studies using a recombinant acidic C-terminal domain of hnRNP-C overexpressed in Escherichia coli demonstrate that protein kinase CK2 phosphorylates hnRNP-C1/C2 at S247, while protein kinase A and several protein kinase C isoforms fail to phosphorylate the isolated domain. These findings demonstrate that the acidic C-terminal domain of hnRNP C1/C2 serves as the site for both basal and stimulated phosphorylation, indicating that this domain may play an important role in the regulation of mRNA binding by hnRNP-C1/C2. PMID- 12564934 TI - Effect of independent variations in fatty acid structure and chain length on lipid polar headgroup composition in Acholeplasma laidlawii B membranes: regulation of lamellar/nonlamellar phase propensity. AB - We have studied the biosynthetic regulation of the membrane lipid polar headgroup distribution in Acholeplasma laidlawii B cells made fatty acid auxotrophic by growth in the presence of the biotin-binding agent avidin to test whether this organism has the ability to coherently regulate the lamellar/nonlamellar phase propensity of its membrane lipids. The addition of various single normal growth supporting exogenous fatty acids to such cell cultures produces fatty acid homogeneous cells in which the hydrocarbon chain length and structure of the fatty acyl chains of the membrane lipids can be independently varied. Moreover, in analyzing our results, we consider the fact that the individual membrane lipid classes of this organism can form either normal micellar, lamellar, or reversed cubic or hexagonal phases in isolation (Lewis, R. N. A. H., and McElhaney, R. N. (1995) Biochemistry 34, 13818-13824). When A. laidlawii cells are highly enriched in one of a homologous series of methyl isobranched, methyl anteisobranched, or omega-cyclohexyl fatty acids, neither the ratio of normal micellar/lamellar nor of inverted cubic or hexagonal/lamellar phase-forming lipids are coherently regulated, and in fact in the former case, the changes in lipid polar headgroup composition observed are generally in a direction opposite to that required to maintain the overall lamellar/nonlamellar phase preference of the total membrane lipids constant when hydrocarbon chain length is varied. Similarly, when lipid hydrocarbon structure is varied at a constant effective chain length, a similar lack of coherent regulation of membrane lipid polar headgroup distribution is also observed, although in this case a weak overall trend in the expected direction occurs. We also confirm our previous finding (Foht, P. J., Tran, Q. M., Lewis, R. N. A. H., and McElhaney, R. N. (1995) Biochemistry 34, 13811-13817) that the ratio of inverted phase-forming monoglucosyl diacylglycerol to the lamellar phase-forming glycolipid diglucosyl diacylglycerol, previously used to estimate membrane lipid phase preference in A. laidlawii A and B, is not by itself a reliable indicator of the overall lamellar/nonlamellar phase propensity of the total membrane lipids of these organisms. Our results indicate that A. laidlawii B lacks a coherent mechanism to biosynthetically regulate the polar headgroup distribution of its membrane lipids to maintain the micellar/lamellar/inverted phase propensity constant in the face of induced variations in either the chain length or the structure of its lipid hydrocarbon chains. Finally, we suggest that the lack of a coherent regulatory mechanism to regulate the overall phase-forming propensity of the total membrane lipids of this organism under these circumstances may result in part from its inability to optimize all of the biologically relevant physical properties of its membrane lipid bilayer simultaneously. PMID- 12564935 TI - Alternative prion structural changes revealed by high pressure. AB - At high temperature, recombinant hamster prion protein (SHaPrP(90-231)) undergoes aggregation and changes from a predominantly alpha-helical to beta-sheet conformation. We then applied high pressure (200 MPa) to the beta-sheet-rich conformation. The aggregation was reversed, and the original tertiary and secondary structures were recovered at ambient pressure, after pressure release. The application of a pressure of 200 MPa thus allowed studying the heat-induced equilibrium refolding in the absence of protein aggregation. Prion protein unfolding as a function of high pressure was also investigated. Simple two-state, reversible unfolding transitions were observed, as monitored by spectral changes in the UV and fluorescence of the hydrophobic probe 8-anilino-1-naphthalene sulfonate. However, these heat- and pressure-induced conformers differed in their unfolding free energy. At pressures over 400 MPa, strong thioflavin-T binding was observed, suggesting a further structural change to a metastable oligomeric structure. PMID- 12564936 TI - Molecular dynamics study of the connection between flap closing and binding of fullerene-based inhibitors of the HIV-1 protease. AB - The complementary spatial relationship between fullerene C(60) and the hydrophobic cavity region of the human immunodeficiency virus (HIV) protease, which houses the active site of the enzyme, has led to the suggestion that fullerene-based derivatives could have potential use as effective HIV protease inhibitors. The ability of such compounds to desolvate the cavity region leads to a strong hydrophobic interaction between the C(60) moiety and residues in the cavity region. In this study, the connection between the motion of the so-called flexible flaps of the cavity and favorable binding of a fullerene-based protease inhibitor is explored using multiple-time scale molecular dynamics simulations and free energy techniques. In addition, the effect of the interaction between the C(60) moiety and the residues in the cavity region on the water content of the cavity is also investigated. Conformational free energy profiles along a suitably chosen flap opening coordinate show a considerable barrier to flap opening in the presence of the inhibitor, while no such barrier exists for the protease alone. This result is interpreted in terms of a strong hydrophobic interaction between the C(60) moiety and the flexible flaps, which cause the latter to close tightly around the inhibitor, thereby expelling water from the cavity and leading to a favorable binding interaction. This interpretation is rationalized by direct analysis of the water content in the cavity in the presence and absence of the inhibitor. PMID- 12564937 TI - Dissection of the EntF condensation domain boundary and active site residues in nonribosomal peptide synthesis. AB - Nonribosomal peptide synthetases (NRPSs) make many natural products of clinical importance, but a deeper understanding of the protein domains that compose NRPS assembly lines is required before these megasynthetases can be effectively engineered to produce novel drugs. The N-terminal amide bond-forming condensation (C) domain of the enterobactin NRPS EntF was excised from the multidomain synthetase using endpoints determined from sequence alignments and secondary structure predictions. The isolated domain was well-folded when compared by circular dichroism to the vibriobactin NRPS VibH, a naturally free-standing C domain. The EntF domain was also fully functional in an assay based on a synthetic small-molecule substrate, seryl N-acetylcysteamine. Active site mutants of the EntF C domain were surprisingly inactive in vitro as compared to their VibH counterparts, yet maintained the overall domain structure. An in vivo assay was developed in the context of the full-length EntF protein to more sensitively probe the activity level of the C domain mutants, and this supported strong effects for the active site mutations. The crucial role of histidine-138 was confirmed by assay of the full-length protein in vitro. These results suggest a strong resemblance of catalysis by the EntF C domain to chloramphenicol acetyltransferase, including an active site organized by an arginine-aspartate salt bridge, a key histidine acting as a general base, and an asparagine instead of a serine stabilizing the proposed tetrahedral intermediate by hydrogen bonding. The precise definition of a functional C domain excised from a NRPS should aid efforts at swapping NRPS domains between assembly lines. PMID- 12564938 TI - Transition state P-glycoprotein binds drugs and modulators with unchanged affinity, suggesting a concerted transport mechanism. AB - The P-glycoprotein multidrug transporter is a plasma membrane efflux pump for hydrophobic natural products, drugs, and peptides, driven by ATP hydrolysis. Determination of the details of the catalytic cycle of P-glycoprotein is critical if we are to understand the mechanism of drug transport and design ways to inhibit it. It has been proposed that the vanadate-trapped transition state of P glycoprotein (Pgp x ADP x V(i) x M(2+), where M(2+) is a divalent metal ion) has a very low affinity for drugs compared to resting state protein, thus leading to binding of substrate on the cytoplasmic side of the membrane and release of substrate to the extracellular medium (or the extracellular membrane leaflet). We have used several different fluorescence spectroscopic approaches to show that isolated purified P-glycoprotein, when trapped in a stable transition state with vanadate and either Co(2+)or Mg(2+), binds drugs with high affinity. For vinblastine, colchicine, rhodamine 123, and doxorubicin, the affinity of the vanadate-trapped transition state for drugs was only very slightly (less than 2 fold) lower than the binding affinity of resting state Pgp, whereas for the modulators cyclosporin A and verapamil and the substrate Hoechst 33342, the binding affinity was very similar for the two states. The drug binding affinity of the ADP-bound form of the transporter was also comparable to that of the unoccupied transporter. These results suggest that release of drug from the transporter during the catalytic cycle precedes formation of the transition state. PMID- 12564939 TI - An Isc-type extremely thermostable [2Fe-2S] ferredoxin from Aquifex aeolicus. Biochemical, spectroscopic, and unfolding studies. AB - Analysis of the genome of the hyperthermophilic bacterium Aquifex aeolicus has revealed the presence of a previously undetected gene potentially encoding a plant- and mammalian-type [2Fe-2S] ferredoxin. Expression of that gene in Escherichia coli has yielded a novel thermostable [2Fe-2S] ferredoxin (designated ferredoxin 5) whose sequence is most similar to those of ferredoxins involved in the assembly of iron-sulfur clusters (Isc-Fd). It nevertheless differs from the latter proteins by having deletions near its N- and C-termini, and no cysteine residues other than those involved in [2Fe-2S] cluster coordination. Resonance Raman, low-temperature MCD and EPR studies show close spectral similarities between ferredoxin 5 and the Isc-Fd from Azotobacter vinelandii. Mossbauer spectra of the reduced protein were analyzed with an S = 1/2 spin Hamiltonian and interpreted in the framework of the ligand field model proposed by Bertrand and Gayda. The redox potential of A. aeolicus ferredoxin 5 (-390 mV) is in keeping with its relatedness to Isc-Fd. Unfolding experiments showed that A. aeolicus ferredoxin 5 is highly thermostable (T(m) = 106 degrees C at pH 7), despite being devoid of features (e.g., high content of charged residues) usually associated with extreme thermal stability. Searches for genes potentially encoding plant type [2Fe-2S] ferredoxins have been performed on the sequenced genomes of hyperthermophilic organisms. None other than the two proteins from A. aeolicus were retrieved, indicating that this otherwise widely distributed group of proteins is barely represented among hyperthermophiles. PMID- 12564940 TI - Phenomenology of self-restraint. AB - Self-restraint is often reported in individuals with mental retardation who show self-injurious behavior (SIB). In this study, the phenomenology and prevalence of self-restraint in individuals showing self-injury and wearing protective devices and those showing self-injury but not wearing protective devices were compared. A high prevalence of self-restraint in the whole sample of individuals showing self injury was identified (67/88, 76.1%), and self-restraint was more prevalent in a group showing self-injury but not wearing protective devices (43/47, 91.5%) than in a group showing self-injury and wearing protective devices (24/41, 58.5%). Individuals not wearing protective devices showed a greater number of topographies of self-restraint than those who did wear them. Results are discussed with reference to the purely topographical definition of self-restraint employed and the potential equivalence of protective devices and self-restraint. PMID- 12564941 TI - Psychotherapy with persons who have mental retardation: an evaluation of effectiveness. AB - Studies of psychotherapy with persons who have mental retardation were examined. Studies conducted during a 30-year period were rated by an expert consensus panel and classified with regard to the nature of the research and outcome and effectiveness domains. We conducted a meta-analysis on a small number of the studies and found a wide range of research designs, types of interventions, and participants. The evaluation also pointed to a moderate degree of change in outcome measures and moderate effectiveness in terms of benefit to clients with mental retardation. We conclude that psychotherapeutic interventions should be considered as part of overall treatment plans for persons with mental retardation. PMID- 12564942 TI - Implicit learning in children and adolescents with mental retardation. AB - The literature on implicit learning in persons with mental retardation is scarce and contradictory with respect to the relationship between degree of intellectual disability and impact of implicit-learning processes on performance. We examined children and adolescents with mild or moderate mental retardation and typically developing children matched on MA with regard to their implicit learning. Individuals with mental retardation modified their behavior after an implicit training procedure in a way similar to MA- or CA-matched controls. The impact of implicit learning did not vary as a function of IQ or age. However, some differences appeared between groups in their explicit remembering of the training conditions. The theoretical implications of these results are discussed. PMID- 12564943 TI - Differential outcomes effect in children and adults with down syndrome. AB - In previous studies, researchers have demonstrated that learning of symbolic relations is facilitated when a particular outcome is associated with each relation to be learned. In the present study, we extend this differential outcomes procedure to children and adults with Down syndrome who had to learn a symbolic conditional discrimination task. Participants showed a better terminal accuracy and a faster learning of the task when the alternative correct responses were each followed by unique different outcomes than when nondifferential outcomes were arranged. These findings confirm that the differential outcomes procedure can be a useful tool to ameliorate discriminative learning deficits and demonstrate the benefits of this procedure for people with Down syndrome. PMID- 12564944 TI - Menstrual discomfort as a biological setting event for severe problem behavior: assessment and intervention. AB - Assessment and intervention approaches for problem behavior need to be extended to deal with biological setting events. To meet assessment needs, we developed a strategy involving four components: interview, archival record review, direct observation, and functional analysis. Data indicated that problem behaviors (aggression, self-injury, and tantrums) were linked to both a biological setting event (menstrual discomfort) and certain discriminative stimuli (task demands). To meet intervention needs, we developed a multicomponent strategy that addressed both the biological context (menstrual discomfort) and the psychosocial context (task demands). Intervention reduced problem behaviors to near-zero levels, maintained for 15 to 22 months across 3 participants. We discuss the extension of our approach to other biological setting events as well as multiple motivational variables. PMID- 12564945 TI - Reinforcer rate effects and behavioral momentum in individuals with developmental disabilities. AB - Behavioral momentum theory states that behavioral resistance to change is positively related to reinforcer rate and independent of response rate under most circumstances. We examined behavioral momentum in humans with developmental disabilities. The experimental procedures were implemented as a computer game. Different rates of positive reinforcement were programmed in two alternating components signaled by distinctive cues. For 10 participants who successfully completed testing, resistance to disruption by an alternate source of reinforcement was greater in the component with the higher reinforcer rate, although the magnitude of the difference varied among individuals. These results confirm reinforcer rate effects consistent with behavioral momentum under laboratory conditions and with the largest number of human beings with developmental disabilities studied to date. PMID- 12564947 TI - The effect of walking aids on balance and weight-bearing patterns of patients with hemiparesis in various stance positions. AB - BACKGROUND AND PURPOSE: Standard and quad canes are often prescribed to patients with hemiparesis, yet their effect on postural control remains unclear. Thus, the objective of this study was to examine the effects of standard and quad canes on postural sway and on weight-bearing patterns in patients with hemiparesis. SUBJECTS: Thirty subjects with a diagnosis of unilateral hemiparesis following a stroke (patient group; mean age=71.2 years, SD=7.0) and 20 age-matched, community dwelling volunteers without hemiparesis (comparison group; mean age=72.1 years, SD=5.2) participated in the study. METHODS: Postural sway and percentage of body weight (%BW) borne by each extremity were measured in 3 positions: with the heels aligned with each other (aligned position) and in staggered foot positions with either the affected or unaffected extremity placed forward (affected FW and unaffected FW positions). All subjects were tested in each position with no cane, a standard cane, and a quad cane. The order of tests was randomized, and analysis of data included use of an analysis of variance and adjusted Tukey-Kramer tests. RESULTS: In both the aligned and unaffected FW positions, postural sway was reduced only with the quad cane. Both types of canes reduced postural sway in the affected FW position; however, the quad cane had a greater effect. An asymmetrical weight distribution between the lower extremities did not change in the patient group across positions, even with walking aids. DISCUSSION AND CONCLUSION: A quad cane appears to be more effective than a standard cane in decreasing postural sway in patients with moderate impairment secondary to hemiparesis during stance. The greatest effect on postural sway occurred when the assistive device was contralateral to the foot placed forward. The use of a cane does not appear to adversely affect the asymmetrical weight-bearing pattern during stance that is characteristic of patients with hemiparesis, even when balance is challenged by decreasing the base of support. PMID- 12564948 TI - A quantitative analysis of research publications in physical therapy journals. AB - BACKGROUND AND PURPOSE: Many physical therapists depend on their professional journals for high-quality evidence. The purpose of this study was to evaluate the rigor of research and review articles in 4 national physical therapy journals. SUBJECTS AND METHODS: All articles in 6 consecutive issues of the Australian Journal of Physiotherapy, Physical Therapy, Physiotherapy, and Physiotherapy Canada, published between January 2000 and June 2001 (N=179), were reviewed. One trained reviewer identified the type and purpose of each article and assessed the rigor of treatment and review articles according to explicit criteria. RESULTS: The majority of articles reviewed were original studies (56%). The majority of the research articles that dealt with human health care (66%) addressed topics that were not directly applicable to the provision of patient care such as measurement topics and studies on subjects without identified pathologies or impairments. Of the 179 journal articles, 19 met the standards for rigor (11%). The majority of these articles dealt with treatment. The pass rate per journal was as follows: Australian Journal of Physiotherapy, 10% (4/42); Physical Therapy, 15% (7/47); Physiotherapy, 12% (4/34); and Physiotherapy Canada, 7% (4/56). DISCUSSION AND CONCLUSION: Because such a small percentage of articles in these professional journals were identified as having direct application to patient care, physical therapists should attempt to access other sources of information to find additional high-quality evidence. A larger sample with a greater number of issues per journal may have yielded different results and indicated different trends, and further research appears to be warranted. PMID- 12564949 TI - Concurrent validity of upper-extremity volume estimates: comparison of calculated volume derived from girth measurements and water displacement volume. AB - BACKGROUND AND PURPOSE: The volume of all limbs can be determined by water displacement methods or calculations derived from girth measurements. The purpose of this study was to determine the concurrent validity of calculated volume and water displacement volume measurements. SUBJECTS: Both upper extremities of 14 women with lymphedema were measured. METHODS: Volumetric measurements were taken with a volumeter, and circumferential measurements were taken with a tape measure. Calculated volume was determined by summing segment volumes derived from the truncated cone formula. Pearson product moment correlations, paired t tests, and linear regression tests were used to assess relative association and absolute differences between calculated and actual volumes. RESULTS: The correlation coefficient for calculated volume versus upper extremity minus fingers (UE-F) water displacement volume was.99. Paired t tests showed differences between calculated volume and UE-F water displacement volume (t=-3.88, mean difference= 95.62 mL), and the linear regression slope was 0.83 with an intercept of 255.28 mL. DISCUSSION AND CONCLUSION: Calculated volume measurements were highly associated with measurements based on water displacement; therefore, clinicians should feel confident in using either calculated volume or water displacement volume. The differences, however, indicated that the measures were not interchangeable. Thus, clinicians should not mix or substitute measurement methods with a single patient or in a single study. PMID- 12564950 TI - Accuracy and reliability of observational gait analysis data: judgments of push off in gait after stroke. AB - BACKGROUND AND PURPOSE: Physical therapists routinely observe gait in clinical practice. The purpose of this study was to determine the accuracy and reliability of observational assessments of push-off in gait after stroke. SUBJECTS: Eighteen physical therapists and 11 subjects with hemiplegia following a stroke participated in the study. METHODS: Measurements of ankle power generation were obtained from subjects following stroke using a gait analysis system. Concurrent videotaped gait performances were observed by the physical therapists on 2 occasions. Ankle power generation at push-off was scored as either normal or abnormal using two 11-point rating scales. These observational ratings were correlated with the measurements of peak ankle power generation. RESULTS: A high correlation was obtained between the observational ratings and the measurements of ankle power generation (mean Pearson r=.84). Interobserver reliability was moderately high (mean intraclass correlation coefficient [ICC (2,1)]=.76). Intraobserver reliability also was high, with a mean ICC (2,1) of.89 obtained. DISCUSSION AND CONCLUSION: Physical therapists were able to make accurate and reliable judgments of push-off in videotaped gait of subjects following stroke using observational assessment. Further research is indicated to explore the accuracy and reliability of data obtained with observational gait analysis as it occurs in clinical practice. PMID- 12564951 TI - Effects of iontophoresis current magnitude and duration on dexamethasone deposition and localized drug retention. AB - BACKGROUND AND PURPOSE: Iontophoresis is a process that uses bipolar electric fields to propel molecules across intact skin and into underlying tissue. The purpose of this study was to describe and experimentally examine an iontophoresis drug delivery model. SUBJECTS AND METHODS: A mechanistic model describing delivery was studied in vitro using agarose gels and was further tested in vivo by evaluation of cutaneous vasoconstriction following iontophoresis in human volunteers. RESULTS: In vitro cathodic iontophoresis at 4 mA and 0.1 mA each delivered dexamethasone/dexamethasone phosphate (DEX/DEX-P) from a 4-mg/mL donor solution to a depth of 12 mm following a 40 mA minute stimulation dosage. Delivery of DEX/DEX-P to at least the depths of the vasculature in humans was confirmed by observation of cutaneous vasoconstriction. This cutaneous vasoconstriction was longer lasting and greater in magnitude when using low current, long-duration (approximately 0.1 mA) iontophoresis compared with equivalent dosages delivered by higher-current, shorter-duration (1.5-4.0 mA) iontophoresis. DISCUSSION AND CONCLUSION: From data gathered with the gel model, the authors developed a model of a potential mechanism of drug depot formation following iontophoresis. The authors believe this drug depot formation to be due to exchange of drug ions for chloride ions as the ionic current carriers. Furthermore, diffusion, not magnitude of current, appears to govern the depth of drug penetration. Although the authors did not address the efficacy of the drug delivered, the results of human experiments suggest that current magnitude and duration should be considered as factors in treating musculoskeletal dysfunctions with iontophoresis using DEX/DEX-P at a concentration of 4 mg/mL. PMID- 12564952 TI - Use of inspiratory strength training to wean six patients who were ventilator dependent. AB - BACKGROUND AND PURPOSE: Patients who are unable to wean from mechanical ventilation (MV) after resolution of critical illness or surgery risk increased morbidity and death and consume a disproportionate amount of intensive care unit resources. Decreased inspiratory muscle strength is often cited as a major factor contributing to prolonged MV. The purpose of this case report is to describe the rationale and application of inspiratory strength training (IST) as an adjunct to lengthen unassisted breathing trials and to ultimately wean patients with chronic mechanical ventilator dependency. CASE DESCRIPTION: Six patients who had been ventilator-dependent for 18 to 221 days (mean of 72 days) after surgery and were diagnosed with "failure to wean" performed low-repetition, high-resistance breathing exercises that were coupled with increasing time off the ventilator. OUTCOMES: All 6 patients were weaned from the ventilator in 9 to 28 days (mean of 17 days). The patients' training pressure increased from a mean of 9.3 cm/H2O to 27.5 cm/H2O, for an increase of 195%. The volitional maximum inspiratory pressure (MIP) increased from a mean of 22.5 cm/H2O to 54 cm/H2O, for a 140% gain in pressure. DISCUSSION: Although it is not clear why the patients appeared to benefit from IST, possible explanations include: (1) addressing inspiratory muscle pump dysfunction, (2) standardization of breathing patterns, 3) routinization of the patients' unassisted breathing trials, and (4) nonspecific training effects. Future research should address these possibilities when attempting to understand the effects of IST in the weaning of patients with chronic ventilator dependency. PMID- 12564954 TI - Expression profile of the transient receptor potential (TRP) family in neutrophil granulocytes: evidence for currents through long TRP channel 2 induced by ADP ribose and NAD. AB - An early key event in the activation of neutrophil granulocytes is Ca(2+) influx. Members of the transient receptor potential (TRP) channel family may be held responsible for this. The aim of the present study is to analyse the expression pattern of TRP mRNA and identify characteristic currents unambiguously attributable to particular TRP channels. mRNA was extracted from human neutrophils, isolated by gradient centrifugation and also by magnetically labelled CD15 antibodies. The presence of mRNA was demonstrated using reverse transcriptase-PCR in neutrophils (controlled to be CD5-negative) as well as in human leukaemic cell line 60 (HL-60) cells, for the following TRP species: the long TRPC2 (LTRPC2), the vanilloid receptor 1, the vanilloid receptor-like protein 1 and epithelial Ca(2+) channels 1 and 2. TRPC6 was specific for neutrophils, whereas only in HL-60 cells were TRPC1, TRPC2, TRPC3, melastatin 1 and melastatin-related 1 found. Patch-clamp measurements in neutrophils revealed non-selective cation currents evoked by intracellular ADP-ribose and by NAD(+). Both these modes of activation have been found to be characteristic of LTRPC2. Furthermore, single-channel activity was resolved in neutrophils and it was indistinguishable from that in LTRPC2-transfected HEK-293 cells. The results provide evidence that LTRPC2 in neutrophil granulocytes forms an entry pathway for Na(+) and Ca(2+), which is regulated by ADP-ribose and the redox state. PMID- 12564953 TI - Regulation of endocytic traffic by Rho GTPases. AB - The members of the Rho subfamily of small GTPases are key regulators of the actin cytoskeleton. However, recent studies have provided evidence for multiple additional roles for these signalling proteins in controlling endocytic traffic. Here we review our current understanding of Rho GTPase action within the endocytic pathway and examine the potential points of convergence with the more established, actin-based functions of these signalling proteins. PMID- 12564955 TI - The aorta and heart differentially express RGS (regulators of G-protein signalling) proteins that selectively regulate sphingosine 1-phosphate, angiotensin II and endothelin-1 signalling. AB - Normal cardiovascular development and physiology depend in part upon signalling through G-protein-coupled receptors (GPCRs), such as the angiotensin II type 1 (AT(1)) receptor, sphingosine 1-phosphate (S1P) receptors and endothelin-1 (ET-1) receptor. Since regulator of G-protein signalling (RGS) proteins function as GTPase-activating proteins for the G alpha subunit of heterotrimeric G-proteins, these proteins undoubtedly have functional roles in the cardiovascular system. In the present paper, we show that human aorta and heart differentially express RGS1, RGS2, RGS3S (short-form), RGS3L (long-form), PDZ-RGS3 (PDZ domain containing) and RGS4. The aorta prominently expresses mRNAs for all these RGS proteins except PDZ-RGS3. Various stimuli that are critical for both cardiovascular development and function regulate dynamically the mRNA levels of several of these RGS proteins in primary human aortic smooth muscle cells. Both RGS1 and RGS3 inhibit signalling through the S1P(1) (formerly known as EDG-1), S1P(2) (formerly known as EDG-5) and S1P(3) (formerly known as EDG-3) receptors, whereas RGS2 and RGS4 selectively attenuate S1P(2)-and S1P(3)-receptor signalling respectively. All of the tested RGS proteins inhibit AT(1)-receptor signalling, whereas only RGS3 and, to a lesser extent, RGS4 inhibit ET(A)-receptor signalling. The conspicuous expression of RGS proteins in the cardiovascular system and their selective effects on relevant GPCR-signalling pathways provide additional evidence that they have functional roles in cardiovascular development and physiology. PMID- 12564956 TI - 5 S rRNA: structure and interactions. AB - 5 S rRNA is an integral component of the large ribosomal subunit in all known organisms. Despite many years of intensive study, the function of 5 S rRNA in the ribosome remains unknown. Advances in the analysis of ribosome structure that have revealed the crystal structures of large ribosomal subunits and of the complete ribosome from various organisms put the results of studies on 5 S rRNA in a new perspective. This paper summarizes recently published data on the structure and function of 5 S rRNA and its interactions in complexes with proteins, within and outside the ribosome. PMID- 12564976 TI - Bispecific antibody conjugates in therapeutics. AB - Bispecific monoclonal antibodies have drawn considerable attention from the research community due to their unique structure against two different antigens. The two-arm structure of bsMAb allows researchers to place a therapeutic agent on one arm while allowing the other to specifically target the disease site. The therapeutic agent can be a drug, toxin, enzyme, DNA, radionuclide, etc. Furthermore, bsMAb may redirect the cytotoxicity of immune effector cells towards the diseased cells or induce a systemic immune response against the target. BsMAb holds great promise for numerous therapeutic needs in the light of: (1) recent breakthroughs in recombinant DNA technology, (2) the increased number of identified disease targets as the result of the completion of human genomic map project, and (3) a better understanding of the mechanism of human immune system. This review focuses on therapeutic applications and production of bsMAb while providing the up-to-date clinical trial information. PMID- 12564977 TI - Drug delivery strategy utilizing conjugation via reversible disulfide linkages: role and site of cellular reducing activities. AB - The first disulfide linkage-employing drug conjugate that exploits the reversible nature of this unique covalent bond was recently approved for human use. Increasing numbers of drug formulations that incorporate disulfide bonds have been reported, particularly in the next generation macromolecular pharmaceuticals. These are designed to exploit differences in the reduction potential at different locations within and upon cells. The recent characterization of a novel redox enzyme in endosomes and lysosomes adds more excitement to this approach. This review focuses on understanding where and how the disulfide bond in the bioconjugate is reduced upon contact with biological milieu, which affects delivery design and the interpretation of the delivery strategies. PMID- 12564978 TI - Effective drug delivery by PEGylated drug conjugates. AB - The current review presents an update of drug delivery using poly(ethylene glycol) (PEG), that focuses on recent developments in both protein and organic drugs. Certainly the past 10 years has resulted in a renaissance of the field of PEG drug conjugates, initiated by the use of higher molecular weight PEGs (M(w)>20,000), especially 40,000 which is estimated to have a plasma circulating t(1/2) of approximately 10 h in mice. This recent resuscitation of small organic molecule delivery by high molecular weight PEG conjugates was founded on meaningful in vivo testing using established tumor models, and has led to a clinical candidate, PEG-camptothecin (PROTHECAN), an ester based prodrug currently in phase II trials. Additional applications of high molecular weight PEG prodrug strategies to amino containing drugs are presented: similar tripartate systems based on lower M(w) PEG and their use with proteins is expounded on. The modification of a benzyl elimination tripartate prodrug specific for mercaptans is presented, and its successful application to 6 mercaptopurine giving a water soluble formulation is discussed. Recent novel PEG oligonucleotides and immunoconjugates are also covered. Clinical results of FDA approved PEGylated proteins are also presented. PMID- 12564979 TI - ATTEMPTS: a heparin/protamine-based triggered release system for the delivery of enzyme drugs without associated side-effects. AB - A prodrug type delivery system based on competitive ionic binding for the conversion of the prodrug to an active drug has been developed for delivery of enzyme drugs without their associated toxic side-effects. This approach, termed "ATTEMPTS" (antibody targeted, triggered, electrically modified prodrug-type strategy), would permit the administration of an inactive drug and then subsequently triggered release of the active drug at the target site. The underlying principle was to modify the enzyme with small cationic species so that it could bind a negatively charged heparin-linked antibody, and the latter would block the activity of the enzyme drug until it reached the target. To provide the enzyme drug with appropriate binding strength to heparin, a cationic poly(Arg)(7) peptide was incorporated onto the enzyme either by the chemical conjugation method using a bifunctional crosslinker or by the biological conjugation method using the recombinant methodology. Methods for drug modification, heparin antibody conjugation, and the prodrug and triggered release features of the "ATTEMPTS" approach are described in detail in this review article. PMID- 12564980 TI - Cellular delivery of peptide nucleic acid (PNA). AB - Peptide nucleic acid (PNA) is a DNA mimic having a pseudopeptide backbone that makes it extremely stable in biological fluids. PNA binds complementary RNA and DNA with high affinity and specificity. These qualities make PNA a leading agent among "third generation" antisense and antigene agents. Unfortunately, fast progress in the exploration of PNA as an experimental and therapeutical regulator of gene expression has been hampered by the poor cellular uptake of PNA. However, a number of transfection protocols for PNA have now been established. These include microinjection, electroporation, co-transfection with DNA, conjugation to lipophilic moieties, conjugation to peptides, etc. Here we give a short introduction to the basic findings on PNA as an antisense and antigene agent in cell-free in vitro systems. This is followed by a comprehensive evaluation of the most interesting literature concerning cellular delivery and the intracellular effect of PNA. Also the current progress as regards using PNA as co-factor in DNA delivery is reviewed. PMID- 12564981 TI - Arginine containing peptides as delivery vectors. AB - Recently several membrane translocalizational signals (MTS) have been identified and applied to various applications. These peptide signals, ranging between nine and 30 amino acid residues in length, have the capability of crossing plasma membrane, in addition to delivering other biological molecules into cells. To date, small molecules, peptides, proteins, oligonucleotides, plasmids and even nanometer-sized particles have been delivered. These MTS sequences vary from hydrophobic to purely hydrophilic, and, surprisingly, all of them are able to penetrate cellular membrane in an energy-independent pathway. Potentially, MTS could be used as delivery vectors for a number of therapeutic agents. In this review, we specifically focus on arginine-containing MTS, and their properties, characteristics, in vitro and in vivo applications are discussed in detail. PMID- 12564982 TI - NLS bioconjugates for targeting therapeutic genes to the nucleus. AB - One of the major steps limiting non-viral gene transfer efficiency is the entry of plasmid DNA from the cytoplasm into the nucleus of transfected cells. Trafficking of nuclear proteins from the cytoplasm into the nucleus through nuclear pore complexes is mediated by the presence of nuclear localization sequences (NLS) on proteins. Viral DNA and RNA also require interaction with cellular machinery for efficient nuclear import. In this article, we review the various strategies used to provide plasmid DNA with nuclear localization sequences, and discuss the possibility of developing efficient gene delivery systems based on these strategies. PMID- 12564983 TI - Genomic islands in Photorhabdus. AB - Genomic islands are responsible for unique aspects of bacterial behavior such as symbiosis and pathogenicity. Photorhabdus luminescens is a pathogen of insects that spends part of its lifecycle in symbiosis with a nematode. Here, we describe novel genomic islands from Photorhabdus that are involved in symbiosis and pathogenicity, and discuss the inter-relationship between virulence factors used against invertebrates and vertebrates. PMID- 12564984 TI - Pathogenesis and host response of Helicobacter pylori. AB - The 5th International Workshop on Pathogenesis and Host Response in Helicobacter Infections was held in Elsinore, Denmark, 4-7 July, 2002. PMID- 12564985 TI - Molecular basis of bacterial virulence and survival within infected hosts and the environment. AB - The EMBO-FEBS lecture course on the 'Molecular Basis of Bacterial Virulence and Survival Within Infected Hosts and in the Environment', organized by Pascale Cossart, Efstathios Gonos and Roberto Kolter, was held on the island of Spetsai, Greece, September 3-13, 2002. PMID- 12564986 TI - Yersinia: an update. AB - The 8th International Symposium on Yersinia was held in Turku, Finland, 4-8 September 2002. PMID- 12564991 TI - Bacterial FHA domains: neglected players in the phospho-threonine signalling game? AB - Forkhead-associated (FHA) domains bind phospho-threonine peptides and are known to mediate phosphorylation-dependent protein-protein interactions in a variety of eukaryotic settings. However, their role in bacterial physiology and signalling has been largely neglected. We have surveyed bacterial FHA domains and discovered that they are implicated in many bacterial processes, including regulation of cell shape, type III secretion, sporulation, pathogenic and symbiotic host bacterium interactions, carbohydrate storage and transport, signal transduction and ethambutol resistance. The way is now open to identify the targets of each FHA domain, and their roles in cellular physiology, and perhaps even to develop novel FHA-blocking antibacterial agents. PMID- 12564992 TI - BSE - a wolf in sheep's clothing? AB - The entire sheep flock in the UK has been threatened with slaughter if BSE is found in farmed sheep, largely on the grounds that an epidemic of BSE in sheep could be harder to contain than was the case for cattle, and that lamb could present a greater risk to consumers than beef. However, identifying BSE in a sheep is not straightforward, because of its similarities to the related disease, scrapie. Here, we review the likelihood that any UK sheep have BSE, how they might have got it, how a case could be identified and what the Government is doing in terms of surveillance and possible control methods. PMID- 12564993 TI - Information overload: assigning genetic functionality in the age of genomics and large-scale screening. AB - As more and more genome sequences are completed, it is becoming increasingly evident that our understanding of the function of most bacterial gene products is lacking. This is frustrating, particularly in the study of pathogens, where an understanding of the role of individual gene products would probably facilitate the development of novel antimicrobials and vaccines. Recently, we devised a technique known as virulence-attenuated pool (VAP) screening to help assign genetic functionality to gene products that the pathogen Vibrio cholerae requires for colonization. This screen and potential new applications of the VAP technique are discussed here. PMID- 12564994 TI - Streptococcal beta-hemolysins: genetics and role in disease pathogenesis. AB - A zone of beta-hemolysis surrounding colonies on blood-agar media is a hallmark phenotypic feature of the pathogens group A Streptococcus (GAS) and group B Streptococcus (GBS). In each case, lysis of red blood cells reflects the action of a potent protein exotoxin. Although these toxins have been the subjects of numerous investigations over the years, their purification and molecular identification have proven elusive. These difficulties reflect the instability of hemolytic activity, as both toxins function only in the context of the bacterial surface or certain high molecular weight 'stabilizer' molecules. This review highlights the recent discoveries of two markedly distinct genetic loci, necessary and sufficient for the beta-hemolytic phenotypes of GAS and GBS, respectively. The generation of isogenic GAS and GBS beta-hemolysin-deficient mutants and their analysis using in vitro and in vivo model systems has shown that both toxins function as virulence factors in the pathogenesis of invasive infections. PMID- 12564995 TI - Environmental regulation of mutation rates at specific sites. AB - Recent studies on bacterial adaptation to stress suggest that bacteria can regulate the generation of mutations at specific sites in response to environmental conditions. Here, we review these findings and discuss the circumstances under which these mechanisms might prove advantageous. PMID- 12564999 TI - Squirreling away at Parkinson's. AB - A compound that triggers hibernation in ground squirrels can also protect brain cells from the lack of oxygen following a stroke. PMID- 12565000 TI - Mistletoe compound enters clinical trials. AB - A drug has entered clinical trials that contains a recombinant version of the main extract from mistletoe, adn holds promise for cancer patients. PMID- 12565001 TI - Bad bugs: good for cancer therapy? AB - Components of Listeria and Escherichia coli have been used to prime the immune system in a novel approach to fighting cancer. PMID- 12565004 TI - Rontgen rays revealed. AB - Raymond C Rowe discusses the history of X-rays and the man who discovered them. PMID- 12565005 TI - Polymers for drug delivery: the debate continues. PMID- 12565006 TI - Non-viral cancer gene therapy--what is best? PMID- 12565007 TI - A new era in cancer therapeutics? AB - Highlights from the 14th EORTC-NCI-AACR Symposium Molecular Targets and Cancer Therapeutics, 19-22 November 2002, in Frankfurt, Germany. PMID- 12565009 TI - Anticancer drugs of tomorrow: apoptotic pathways as targets for drug design. AB - Apoptosis or programmed cell death is a set of ordered events that enables the selective removal of cells from tissue and is essential for homeostasis and proper function of multicellular organisms. Components of this signaling network, which include ligands, such as CD95, tumor necrosis factor (TNF) and TNF-related apoptosis-inducing ligand, as well as downstream molecules, such as caspases, Bcl 2 family members, and inhibitor-of-apoptosis proteins, which trigger and regulate apoptosis, are crucial targets for conventional drug development and gene therapy of cancer and other diseases. Here, we focus on apoptotic pathways and propose new potential molecular targets that could prove effective in controlling cell death in the clinical setting. PMID- 12565010 TI - Nuclear transport as a target for cell growth. AB - The function of many key proteins and transcription factors involved in cell growth can be regulated by their cellular localization. Such proteins include the tumor suppressor p53 and the nuclear factor kappaB. Although the idea of trapping such proteins in either the nucleus or cytoplasm has been introduced as a potential therapeutic target, only two nuclear transport inhibitors have been reported. Here, we explore the roles of small-molecule inhibitors that cause target proteins to sequester in either the nucleus or cytoplasm. Methods of artificially targeting proteins to the nucleus or cytoplasm using peptide aptamer technology are also discussed. PMID- 12565011 TI - Nonleadlikeness and leadlikeness in biochemical screening. AB - Biochemical assays have largely supplanted functional biological assays as drug screening tools in the early stages of drug discovery. The de-selection of compounds that are 'nonleadlike' binders (and bonders) and the proactive selection of those compounds that are 'leadlike' in their binding to the target are vital components of the screening effort. The physiochemical properties of leadlikeness and the surprising differences between those properties and the now classical definitions of druglikeness are becoming apparent. PMID- 12565014 TI - Effect of tenoxicam on fracture healing in rat tibiae. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs have been implicated in the development of delayed unions and nonunion after fractures in animal models. Previous investigations have identified two important factors as determinants of delayed fracture healing: early drug administration and a dose-dependent effect. OBJECTIVE: The purpose of this investigation was to study the effect of tenoxicam, a nonsteroidal anti-inflammatory drug, on the fracture healing process in rat tibiae. METHODS: Fifty-eight Wistar rats were randomly divided in four groups (I, II, III, and IV). Group I (control group, n=12) was given 0.1ml saline solution per day intramuscularly. Groups II (n=12), III (n=12), and IV (n=12) were administered 10mg per kg per day of tenoxicam intramuscularly. Administration of substances was begun on a week before to 48h after the fracturing procedure and continued during the entire experiment. Callus formation was studied histologically and histomorphologically, using light microscopy. In addition, a histologic grading based on the morphologic stage of fracture healing was carried out at 4 weeks, according to the criteria proposed by Allen et al. RESULTS: There was a significant difference in treatment effect between Group I (saline solution) and Groups II, III, and IV (tenoxicam) (P=0.07). Histologically and histomorphologically, there were qualitative and quantitative delay in callus formation at all tenoxicam groups. This was more pronounced the earlier the nonsteroidal anti-inflammatory drug was started, although no significant difference could be detected between Groups II, III, and IV (P>(alpha=10%)). Four weeks after fracture, Group I (n=3) showed complete osseous union, Groups II (n=3) and III (n=3), complete cartilaginous union, and Group IV (n=3), incomplete osseous union, according to Allen et al. By using this rating scale, the difference between control and drug-treated groups was statistically significant (P<0.1). CONCLUSION: Under studied conditions, this investigation shows that administration of tenoxicam intramuscularly delays fracture healing process in rat tibiae. These results suggest the hypothesis that early drug administration may delay bone healing after experimental fractures in animals, although it could not be detected statistically significant. PMID- 12565015 TI - The management of periprosthetic femoral fractures around hip replacements. AB - Periprosthetic femoral fractures are increasing in frequency and in complexity. They occur intra-operatively, or post-operatively, when they are frequently associated with loosening, with or without osteolysis. Periprosthetic femoral fractures usually occur at low energy levels, either after falls or spontaneously during activities of daily living. At present, the Vancouver classification system probably comes closest to the ideal, as it considers the fracture configuration, the stability of the implant and the quality of the bone stock. When the stem is stable, open reduction and internal fixation is suggested. Several authors have used strut grafts for the treatment of periprosthetic femoral fractures, with good results. If the stem is loose, or revision is indicated for other reasons, it is generally advisable to by-pass the most distal fracture line with a longer stem prosthesis by at least two femoral diameters. Augmentation of this intra-medullary fixation with an external cortical strut to improve rotational stability and/or internally with impaction allografting to compensate for bone defects is also advisable. Vigilant post-operative clinical and radiological assessment following total hip replacement should identify those with recurrent dislocation, loosening, subsidence and osteolysis. These patients are at greatest risk of developing femoral periprosthetic fractures. PMID- 12565016 TI - Dall-Miles plates for periprosthetic femoral fractures. A critical review of 16 cases. AB - Fourteen patients with 16 periprosthetic femoral fractures around hip replacement were treated with Dall-Miles plates between June 1996 and February 2000. There were 10 Vancouver B3, three B1 and three type C fractures. In addition to a Dall Miles plate, two of the fractures (one B3 and one C type) were also stabilised with one strut graft and nine B3 fractures were revised with impaction grafting. Of the three B1 fractures treated with plates, two failed through fracture of the plate. A further two patients with B3 fractures treated with plates also failed with fracture of the plate. Failure of these plates occurred within 6 months of surgery. All non-unions and fixation failures in this series were in cases where the femoral component did not bypass the most distal fracture line by at least two cortical diameters. The Dall-Miles plates and cable system alone is insufficient for the treatment for periprosthetic femoral fractures. It must be supplemented with additional intramedullary or extramedullary fixation. PMID- 12565017 TI - The use of a locking custom contoured blade plate for peri-articular nonunions. AB - A retrospective chart and radiographic review was performed of 37 operative procedures over 7 years period for peri-articular nonunions. This was a consecutive cohort from a single orthopedic trauma service of a Level 1 center. Nonunions of the tibia and humerus were treated with blade plates fashioned from standard compression plates. Twenty-nine operative sites were treated successfully. The average age was 45.0 years; the average follow-up being 5.3 years. The average number of operations before blade plate was 4.0 (range 1-16). Sixteen blade plates were performed in patients with a diagnosis of clinical infection and all progressed to union and resolution of infection. Seven tissue transfers were used as adjunctive therapy. Five other patients with blade plate surgery had persistent nonunion, of which four united after a second custom contoured blade plate procedure. This study illustrates that surgeon contoured blade plates are an option for peri-articular nonunions even in the presence of infection. This technique resulted in a high union rate and a low complication rate compared to other options. PMID- 12565018 TI - Results of dynamic condylar screw for subtrochanteric fractures. AB - A consecutive series of 58 patients, treated with the dynamic condylar screw (DCS) for subtrochanteric fractures were retrospectively reviewed. The mechanism of injury was low-energy in 47 cases and high-energy in 11 cases. Five patients died before fracture healing. Ten out of 11 young patients, (nine with high energy injuries), united primarily. In contrast, the primary union rate in the elderly was 74%. Implant failure was seen in 11 cases (20%). Restricted weight bearing status post-operatively was associated with significantly lower incidence of implant failure (P<0.05, chi(2)-test). The use of bone graft was not related to fracture union (P>0.5). After revision surgery the overall union rate in the elderly was 90%. Using the DCS, the results were good in the young patients with high-energy injuries. However, the implant failure rate is high in elderly patients who suffer low-energy fractures. The DCS should not be used if weight bearing cannot be minimized in this group. PMID- 12565019 TI - The use of a dynamic condylar screw and biological reduction techniques for subtrochanteric femur fracture. AB - A study was designed to examine the outcomes of patients with closed comminuted subtrochanteric femoral fractures fixed with a dynamic condylar screw (DCS) and using biological (indirect) reduction techniques at a tertiary referral centre.Thirty-one consecutive patients with a mean age of 32.6 years, who sustained subtrochanteric femoral fractures, were treated with this method. Twenty-seven cases (87.2%) sustained these fractures either from traffic crashes or falls from height and the remaining four had a simple fall. Average follow-up was 3 years (range 14-65 months). Patients were assessed clinically and radiographically with regards to fracture classification, operating time, blood loss, time of union, malunion and other complications. Union was achieved in all cases (100%), with full-weight bearing after an average of 4.9 months. The average operating time was 2h and blood loss averaged 430 ml. There was only one case of superficial infection, which settled with local debridement and antibiotics. Malunion was seen in 2 cases out of 31 (6.4%) without the need for further surgery. In conclusion, use of biological (indirect) reduction techniques instead of anatomic, open reduction has proven to be successful, especially in comminuted subtrochanteric fractures. The early appearance of callus avoids the need for primary cancellous bone grafting, emphasising the importance of preserving biology of the fracture fragments. PMID- 12565020 TI - Necrosis of the femoral head after fixation of trochanteric fractures with Gamma Locking Nail. A cause of late mechanical failure. AB - Cut-out of the lag screw after fixation of trochanteric fractures with intramedullary devices like the Gamma Locking Nail is a well documented cause of late mechanical failure. Avascular necrosis of the femoral head after trochanteric fractures is an uncommon complication and several causes have been described. As far as we know, the relation of the mechanical failure of fracture fixation and avascular necrosis of the femoral head in trochanteric fractures has never been described. We present a study of six consecutive patients who underwent arthroplasty after cut-out of the lag screw fixation of a trochanteric fracture with Gamma Locking Nail. All six femoral heads were studied histologically and a clinical retrospective study was also performed. In all cases, we found homogeneous avascular necrosis of the femoral head; in all but one, revascularisation was present in every area examined. We suggest that after fixation of these trochanteric fractures with the Gamma Locking Nail, there is a transitory ischaemia, if the revascularisation is not enough to provide a strong support for the lag screw, it may result in late mechanical failure in these fractures. PMID- 12565021 TI - Indirect reduction and bridge plating of supracondylar fractures of the femur. AB - Between October 1995 and December 1998, 16 patients with A-O type A3, C2 or C3 supracondylar femoral fractures were treated by open reduction and internal fixation using indirect reduction and bridge plating. Seven (44%) patients had open fractures. The patients were followed for a mean of 46 months (range 24-71). All fractures healed. The average time for fracture healing was 18.5 weeks (range 12-28). Four primary bone grafts and three secondary bone grafts were performed. By using the modified Schatzker rating scale, the result of 13 patients (81%) were rated as excellent or good. Complications included two implant failures that were due to full weight bearing before bone healing, and one superficial delayed wound healing. No deep infections were found in this series. The open fracture group needed longer time to heal and had a higher rate of receiving bone grafts. We conclude that indirect reduction and bridge plating with a 95 degrees dynamic condylar screw (DCS) or condylar blade plate can produce favourable results for complex distal femoral fractures. We suggest primary bone grafts or early secondary bone grafts for comminuted open fractures using an indirect reduction technique. PMID- 12565023 TI - Tibial fractures treated with circular fixation: does the use of olive wires at the fracture site improve healing? AB - Clinical experience and published studies suggest that oblique fractures of the tibia are associated with delayed healing and non-union. Experimental studies have attributed this to increased shear at the fracture site. We have adopted the practice of using supplementary olive wires to reduce shear when using circular fixation for these fractures. A complete cohort of 54 oblique tibial fractures treated with the Sheffield Ring Fixator (Orthofix, Verona) was reviewed to elucidate the effect of using additional olive wires on fracture healing/treatment times. Fifty patients were studied in the final analysis. With low-energy injuries, the use of olive wires reduced treatment times significantly (no olives: 37 weeks, olives: 22 weeks, P<0.05), although this was not seen with higher energy injuries (no olives: 44 weeks, olives: 39 weeks, P=NS). There was no evidence of additional complications related to their use. We recommend the use of additional olive wires in the circular fixation of these difficult fractures. PMID- 12565022 TI - An injectable calcium phosphate cement as a bone-graft substitute in the treatment of displaced lateral tibial plateau fractures. AB - In the treatment of displaced tibial plateau fractures, filling the resultant metaphyseal defect is essential for maintaining reduction. This study reports on 14 patients with an average age of 53 (range 34-63) years who sustained such a fracture and whose tibial defect was augmented with an injectable calcium phosphate cement under arthroscopic and fluoroscopic control. Eight fractures were treated with subcortical percutaneous screw fixation, plates were used in four cases and two fractures were treated with bone cement only. Full weight bearing was allowed after 6-12 weeks. At the last follow-up which averaged 28 (range 18-47) months, the reduction achieved at the index operation was not altered in any of the patients except one. Most patients recorded little or no pain with an average VAS score of 1.2 (0-3.4). Ten patients had no limitation of their walking distance. Patient satisfaction was good or excellent in 12 cases. Flexion averaged 140 degrees (range 130-150 degrees ) and extension was unlimited in 13 patients. The Lysholm knee score averaged 80 and the Knee Society score 180 (range 127-195). In our opinion the injectable calcium phosphate cement used here is a promising alternative for filling metaphyseal defects in the treatment of displaced tibial plateau fractures. PMID- 12565024 TI - Ilizarov wire tensioning and holding methods: a biomechanical study. AB - AIM: To investigate and compare two specific methods of Ilizarov wire tensioning and hold. METHOD: This study utilised the vertical Hounsfield test machine H25KS, a stress/strain device with a load-cell linked to a computer program. Firstly, the department's present mechanical tensioners were assessed. Secondly, the method of twisting the three designs of wire holding bolts to achieve wire tension, as described by Ilizarov, was assessed. These bolts are described as "cannulated", "slotted" and "Russian" (which are hexagonal headed with a slot down one side). RESULTS: The mechanical tensioners were found to be accurate, if inefficient, with a maximum producible tension of 1330N. The most effective bolts for creation of tension were the Russian, which produced mean wire tensions of 785N at 45 degrees; 1200N at 90 degrees; 1695N at 135 degrees. The cannulated and slotted bolts regularly broke the wires at 90 degrees twist. CONCLUSION: The findings demonstrated two effective methods. The present tensioners were found to be inefficient when compared to the simple twisting of the wire holding bolts, which created equivalent tensions with ease and are capable of producing greater wire tensions. The Russian bolts are recommended for use when wire tension is created by bolt twisting. PMID- 12565025 TI - The biomechanics of wire fixation in the Ilizarov system. AB - The purpose of this study was to establish the optimal fixation of tensioned wires to the frame construct in the Ilizarov system. The usual torque to which the fixation bolts were tightened in clinical practice was established by serial testing of orthopaedic surgeons' work in our unit. The force required to produce wire slippage from the different types of wire fixation bolts, tightened to a predetermined torque, was measured using a testing rig. Analysis of the usual torque to which bolts were tightened in clinical practice, revealed values in the range of 5-10Nm. The load required to cause failure of the tensioned wire varied considerably depending on the bolt configuration used and the applied torque. In clinical practice, wires are tensioned using a dynamometer to a variety of loads, depending on the clinical situation, up to 1275N. In applying multiple wires across a single ring, as is normal practice, these loads may be increased still further. The total load transmitted by single wires in weightbearing may be up to 2000N. Utilising the results of our work has enabled us to choose, where practically possible, the optimal bolt configuration for wire fixation. In addition, this work has revealed that in order to withstand the loads seen in clinical practice, wire fixation bolts should be tightened to at least 10Nm. Post operatively, we now tighten all our wire fixation bolts to 10Nm, using a calibrated torque wrench. PMID- 12565026 TI - Assessment of the efficacy of Proguard RR-2 radio-protective gloves during forearm manipulation. AB - The hazards of ionising radiation are well known and precautions, such as lead aprons and thyroid shields are routinely used. Orthopaedic surgeon's hands are at particular risk from direct and scatter radiation, when manipulating forearm fractures, due to the proximity of the image intensifier. The use of lead gloves has been recommended in the literature but are seldom employed. Proguard RR-2 gloves provide similar tactile sensitivity to double gloves and are claimed by the manufacturer to provide up to 55% protection in vitro at a direct beam energy level of 60 kV. This claim was tested in a clinical setting. The gloves were worn during forearm manipulations and the radiation dose measured using thermoluminescent dosimeters (TLDs). The results demonstrated a radiation attenuation of 60-64%. These gloves appear to achieve a good compromise between protection and sensitivity and should be included in routine protection against ionising radiation during MUA. PMID- 12565027 TI - A painful tibial union. PMID- 12565028 TI - Longspurs and snow buntings: phylogeny and biogeography of a high-latitude clade (Calcarius). AB - Using complete cytochrome b sequence data, we determined that the genus Calcarius, as presently recognized, is paraphyletic. Calcarius plus Plectrophenax form a highly supported clade composed of two subclades, a "snow bunting" clade comprised of Plectrophenax plus Calcarius mccownii (formerly in the monotypic genus Rhynchophanes), and a "collared" longspur clade of Calcarius lapponicus, ornatus, and pictus. Contrary to conventional thought, Calcarius is not phylogenetically close to either Calamospiza or Emberiza. Unlike these two genera, the taxonomic affinities of Calcarius appear to lie outside of the sparrow (tribe Emberizini) assemblage. Calcarius appears to be a relatively old songbird lineage, originating between 4.2 and 6.2 million years ago. Within Calcarius, pictus and ornatus form a closely related sister pair (2.9% divergent), as do Calcarius nivalis and hyperboreus (0.18% divergent). The group (Calcarius, sensu lato) is inferred to have its origins at relatively high latitudes in the New World. PMID- 12565029 TI - Phylogeny of the Dipsacales s.l. based on chloroplast trnL-F and ndhF sequences. AB - Sequences of the chloroplast trnL-F region and 3(') end ndhF gene were used to elucidate phylogenetic relationships and the delimitation of families within Dipsacales s.l. Parsimony analyses of individual and combined data were conducted using maximum parsimony method. The most parsimonious tree based on combined trnL F and 3(') end ndhF data set recognizes seven major clades of Dipsacales s.l. with the following relationships: Apiales (Adoxaceae ((Diervillaceae, Caprifoliaceae s.str.) (Linnaeaceae (Morinaceae (Dipsacaceae, Valerianaceae))))). Both Sambucus and Viburnum have close relationships with Adoxaceae, supporting their inclusion in this family. Caprifoliaceae s.l. (excluding Sambucus and Viburnum) is polyphyletic, and comprises three clades or families, i.e., Linnaeaceae (Abelia, Dipelta, Kolkwitzia, and Linnaea), Diervillaceae (Weigela and Diervilla) and Caprifoliaceae s.str. (Heptacodium, Leycesteria, Lonicera, Symphoricarpos, and Triosteum). This study focuses on the systematic position of Heptacodium, Triplostegia, and Morinaceae; and suggests that Heptacodium is closely related to the other Caprifoliaceae s.str.; Triplostegia is a sister to Dipsacaceae; Morinaceae, which has an affinity with Dipsacaceae, is possibly a sister group with Dipsacaceae-Valerianaceae clade. Our results are highly congruent with those of and. PMID- 12565030 TI - Rapid evolutionary divergences in reef fishes of the family Acanthuridae (Perciformes: Teleostei). AB - A phylogenetic analysis of the sugeonfish family Acanthuridae was conducted to investigate: (a) the pattern of divergences among outgroup and basal ingroup taxa, (b) the pattern of species divergences within acanthurid genera, (c) monophyly in the genus Acanthurus, and (d) the evolution of thick-walled stomach morphology in the genera Acanthurus and Ctenochaetus. Fragments of the 12S, 16S, t-Pro, and control region mitochondrial genes were sequenced for 21 acanthurid taxa (representing all extant genera) and four outgroup taxa. Unweighted parsimony analysis produced two optimal trees. Both of these were highly incongruent with a previous morphological phylogeny, especially with regard to the placement of the monotypic outgroups Zanclus and Luvarus. The maximum likelihood tree and the morphological phylogeny were not significantly different and the conflicting branches were very short. Split decomposition analysis identified conflict in the placement of long basal branches separated by short internodes, providing further evidence that long branch attraction is an important cause of disagreement between molecular and morphological trees. Parametric bootstrapping rejected hypotheses of monophyly of: (a) the genus Acanthurus and (b) a group containing representatives of Acanthurus/Ctenochaetus with thick-walled stomachs. The branching pattern of the likelihood and split decomposition trees indicates that evolution in the acanthurid clade has involved at least three periods of intense speciation. PMID- 12565031 TI - Phylogenetic relationships of elopomorph fishes inferred from mitochondrial ribosomal DNA sequences. AB - The superorder Elopomorpha, a grouping which includes all teleost fishes that possess a specialized leptocephalous larva [true eels (Anguilliformes), gulpers and bobtail snipe eels (Saccopharyngiformes), bonefishes, spiny eels, and halosaurs (Albuliformes, including Notacanthiformes), ladyfishes and tarpons (Elopiformes, including Megalopiformes)] comprises >800 species for which phylogenetic relationships are poorly understood. In the present study, we analyzed mitochondrial DNA sequences in segments of the 12S and 16S rRNA genes in 33 elopomorph taxa encompassing all of the previously proposed orders, and 9 of the 15 currently recognized families of the Anguilliformes, as well as outgroup representatives from the superorders Osteoglossomorpha (nine species) and Clupeomorpha (three species), to develop phylogenetic hypotheses based on distance and parsimony methods. Both methods failed to support the monophyly of the Elopomorpha, casting doubt on the validity of the leptocephalus as an elopomorph synapomorphy. The orders Elopiformes, Albuliformes, and Anguilliformes, however, were resolved as monophyletic assemblages. Parsimony analysis supported the separation of the Anguilliformes into two groups (primitive and advanced) based on the presence of divided versus fused frontal bones. In addition, the molecular data indicated a close affinity of the anguilliform Thalassenchelys coheni (incertae sedis), known only from the leptocephalus, with the family Serrivomeridae. The implications of these data as regards the evolution of the elopomorph assemblage are discussed. PMID- 12565032 TI - Molecular phylogenetic evidence refuting the hypothesis of Batoidea (rays and skates) as derived sharks. AB - Early morphological studies regarding the evolutionary history of elasmobranchs suggested sharks and batoids (skates and rays) were respectively monophyletic. More modern morphological cladistic studies, however, have tended to suggest that batoids are derived sharks, closely related to sawsharks and angelsharks, a phylogenetic arrangement known as the Hypnosqualea hypothesis. Very few molecular studies addressing interordinal relationships of elasmobranchs have been published; the few that do exist, are very limited in terms of both taxon representation and/or aligned sequence positions, and are insufficient to answer the question of whether batoids are derived sharks. The purpose of this study was to address this issue with more complete taxon representation, concomitant with a reasonable number of aligned sequence positions. The data set included a 2.4-kb segment of the mitochondrial 12S rRNA-tRNA valine-16S rRNA locus, and in terms of taxa, representatives of two orders of Batoidea, at least one representative of all orders of sharks, and as an outgroup, the widely recognized sister group to elasmobranchs-Holocephali. The results provide the first convincing molecular evidence for shark monophyly and the rejection of the Hypnosqualea hypothesis. Our phylogenetic placement of batoids as a basal elasmobranch lineage means that much of the current thinking regarding the evolution of morphological and life history characteristics in elasmobranchs needs to be re-evaluated. PMID- 12565033 TI - Phylogeography of the Madeiran endemic lizard Lacerta dugesii inferred from mtDNA sequences. AB - Partial sequences from two mitochondrial DNA genes, cytochrome b and 12S rRNA, were used to assess the phylogenetic relationships of populations of Lacerta dugesii from the volcanic Atlantic islands of Madeira, the Desertas, Porto Santo, and the Selvagens. All four-island groups are genetically distinguishable and populations within each contain similar degrees of genetic diversity. Molecular clock estimates suggest that the islands were colonized much later after their emergence compared to other Atlantic islands, possibly due to their greater geographical isolation. Mismatch analysis of all populations is consistent with exponential growth, as expected after colonization of empty niches. The Selvagens contain genetic substructuring between the islets. PMID- 12565034 TI - Dramatically elevated rate of mitochondrial substitution in lice (Insecta: Phthiraptera). AB - Few estimates of relative substitution rates, and the underlying mutation rates, exist between mitochondrial and nuclear genes in insects. Previous estimates for insects indicate a 2-9 times faster substitution rate in mitochondrial genes relative to nuclear genes. Here we use novel methods for estimating relative rates of substitution, which incorporate multiple substitutions, and apply these methods to a group of insects (lice, Order: Phthiraptera). First, we use a modification of copath analysis (branch length regression) to construct independent comparisons of rates, consisting of each branch in a phylogenetic tree. The branch length comparisons use maximum likelihood models to correct for multiple substitution. In addition, we estimate codon-specific rates under maximum likelihood for the different genes and compare these values. Estimates of the relative synonymous substitution rates between a mitochondrial (COI) and nuclear (EF-1alpha) gene in lice indicate a relative rate of several 100 to 1. This rapid relative mitochondrial rate (>100 times) is at least an order of magnitude faster than previous estimates for any group of organisms. Comparisons using the same methods for another group of insects (aphids) reveals that this extreme relative rate estimate is not simply attributable to the methods we used, because estimates from aphids are substantially lower. Taxon sampling affects the relative rate estimate, with comparisons involving more closely related taxa resulting in a higher estimate. Relative rate estimates also increase with model complexity, indicating that methods accounting for more multiple substitution estimate higher relative rates. PMID- 12565035 TI - Xantusiid "night" lizards: a puzzling phylogenetic problem revisited using likelihood-based Bayesian methods on mtDNA sequences. AB - Contentious issues in Night Lizard (Xantusiidae) evolution are revisited using Maximum Likelihood-based Bayesian methods and compared with results from Neighbor Joining and Maximum Parsimony analyses. Fragments of three mitochondrial genes, the 12S and 16S ribosomal genes, and the cytochrome b gene, are sampled across an ingroup composed of seven xantusiid species and a 12-species outgroup chosen to bracket ancestral states for six additional clades of scleroglossan lizards. Our phylogenetic analyses afford robust support for the following conclusions: Xantusiidae is part of Scincomorpha, rather than being allied with Gekkota; Lepidophyma is sister to Xantusia, rather than to Cricosaura; Xantusia riversiana is imbedded within, rather than being sister to, other Xantusia species; and rock morph Xantusia are not closely related to one another. Convergence related to retarded rates of growth and development, or to physical constraints imposed by living in rock crevices, may be responsible for much of the character discordance underlying conflicts in xantusiid phylogeny. Fossil-calibrated Maximum Likelihood based divergence time estimates suggest that although the xantusiid stem may have originated in the Mesozoic, the crown clade is exclusively Tertiary in age. Thus, the clade including extant Cricosaura does not appear to have been extant during the K-T boundary bolide impact, as has been suggested. Moreover, our divergence time estimates indicate that the xantusiid island endemics, Cricosaura typica on Cuba and Xantusia riversiana on the California Channel Islands, arrived via dispersal rather than vicariance, as previously proposed. PMID- 12565036 TI - Repeatability of clades as a criterion of reliability: a case study for molecular phylogeny of Acanthomorpha (Teleostei) with larger number of taxa. AB - Although much progress has been made recently in teleostean phylogeny, relationships among the main lineages of the higher teleosts (Acanthomorpha), containing more than 60% of all fish species, remain poorly defined. This study represents the most extensive taxonomic sampling effort to date to collect new molecular characters for phylogenetic analysis of acanthomorph fishes. We compiled and analyzed three independent data sets, including: (i) mitochondrial ribosomal fragments from 12S and 16s (814bp for 97 taxa); (ii) nuclear ribosomal 28S sequences (847bp for 74 taxa); and (iii) a nuclear protein-coding gene, rhodopsin (759bp for 86 taxa). Detailed analyses were conducted on each data set separately and the principle of taxonomic congruence without consensus trees was used to assess confidence in the results as follows. Repeatability of clades from separate analyses was considered the primary criterion to establish reliability, rather than bootstrap proportions from a single combined (total evidence) data matrix. The new and reliable clades emerging from this study of the acanthomorph radiation were: Gadiformes (cods) with Zeioids (dories); Beloniformes (needlefishes) with Atheriniformes (silversides); blenioids (blennies) with Gobiesocoidei (clingfishes); Channoidei (snakeheads) with Anabantoidei (climbing gouramies); Mastacembeloidei (spiny eels) with Synbranchioidei (swamp-eels); the last two pairs of taxa grouping together, Syngnathoidei (aulostomids, macroramphosids) with Dactylopteridae (flying gurnards); Scombroidei (mackerels) plus Stromatoidei plus Chiasmodontidae; Ammodytidae (sand lances) with Cheimarrhichthyidae (torrentfish); Zoarcoidei (eelpouts) with Cottoidei; Percidae (perches) with Notothenioidei (Antarctic fishes); and a clade grouping Carangidae (jacks), Echeneidae (remoras), Sphyraenidae (barracudas), Menidae (moonfish), Polynemidae (threadfins), Centropomidae (snooks), and Pleuronectiformes (flatfishes). PMID- 12565037 TI - Pleistocene effects on the European freshwater fish fauna: double origin of the cobitid genus Sabanejewia in the Danube basin (Osteichthyes: Cobitidae). AB - Biogeographical hypotheses of European freshwater fishes were inferred using phylogeographic analysis of the complete cytochrome b and ATP synthase 8 and 6 mitochondrial genes (1982bp). To test the relative importance of drainage origin versus Pleistocene glaciations in the origin of primary freshwater fishes in Europe, we reconstructed the phylogenetic relationships of the genus Sabanejewia which is distributed in European waters. The phylogenetic relationships recovered for the genus Sabanejewia (n=75) provide support for the monophyly of six main evolutionary mtDNA lineages: Sabanejewia larvata, Sabanejewia romanica, Sabanejewia aurata/Sabanejewia caucasica, Sabanejewia kubanica, Sabanejewia baltica, and the Danubian-Balkanian complex. The Caucasian-Caspian mtDNA lineages, S. kubanica, S. aurata/S. caucasica, and the Northern European S. baltica represents the sister group of the Danubian-Balkanian complex mtDNAclade, supporting a Caucasian-Northern European origin of most of mtDNA lineages of the Central European freshwater fish fauna. The mtDNA divergence observed between the Danubian Sabanejewia species is too dissimilar to support their contemporary origin. Rather, the mtDNA data suggest that the Danubian Sabanejewia lineages most likely have a double origin, indicating that the European Sabanejewia lineages have experienced different historical processes for the following reasons. First, the origin of the S. larvata and S. romanica mtDNA clades predates the origin of the Danubian-Balkanian complex, and our results showed that the completion of the Alps and the origin of the Danube drainage seem to have promoted the speciation of the earliest Sabanejewia clades in the Miocene. Second, small genetic distances and the geographical pattern found within the Danubian-Balkanian complex clade indicate that the lineages included in this clade spread recently across the Danube and Greek river drainages. The inclusion of the S. balcanica species within all mtDNA lineages suggests that cyclical cold periods during the Pleistocene glaciations have favoured its rapid expansion and genetic homogenisation across Central European and Greek waters. PMID- 12565038 TI - Evolution of Mexican Bursera (Burseraceae) inferred from ITS, ETS, and 5S nuclear ribosomal DNA sequences. AB - I reconstructed a phylogeny of 66 species and varieties of Bursera and 9 outgroup species using sequences of the internal transcribed spacer region (ITS), the 5S non-transcribed region (5S-NTS), and the external transcribed region (ETS) of nuclear ribosomal DNA. This study extends a previously proposed parsimony-based phylogenetic study that used the ITS sequences of 57 Bursera species and five outgroups. Parsimony and maximum likelihood methods were used to infer the phylogeny in this new study. Analyses of the combined data sets largely confirmed the phylogenetic relationships proposed by the previous molecular study but generated a considerably more robust topology. The new phylogenies corroborate the monophyly of the genus, and its division into the two monophyletic subgenera or sections, Bursera and Bullockia. The current analyses also identify four main groups of species in section Bursera, and two in section Bullockia, confirming some of the previously proposed groups based on fruit, flower, and leaf morphology. One previously problematic species B. sarcopoda, which has sometimes been placed in Commiphora, is shown to belong in Bursera. Another controversial species, Commiphora leptophloeos, which was thought to belong to Bursera, falls within Commiphora. PMID- 12565039 TI - Climatic oscillations triggered post-Messinian speciation of Western Palearctic brown frogs (Amphibia, Ranidae). AB - Oscillating glacial cycles over the past 2.4 million years are proposed to have had a major impact on the diversity of contemporary species communities. We used mitochondrial and nuclear DNA sequence data to infer phylogenetic relationships within Western Palearctic brown frogs and to test the influence of Pliocene and Pleistocene climatic changes on their evolution. We sequenced 1976bp of the mitochondrial genes 16S rRNA and cytochrome b and of the nuclear rhodopsin gene for all current species and subspecies. Based on an established allozyme clock for Western Palearctic water frogs and substitution rate constancy among water frogs and brown frogs, we calibrated a molecular clock for 1425bp of the 16S and rhodopsin genes. We applied this clock to date speciation events among brown frogs. Western Palearctic brown frogs underwent a basal post-Messinian radiation about 4 million years ago (mya) into five major clades: three monotypic lineages (Rana dalmatina, Rana latastei, Rana graeca), an Anatolian lineage, and a lineage comprising Rana italica, Rana arvalis, and all Iberian taxa. Polytypic lineages radiated further in concordance with the onset of climatic oscillations ca. 3.2, 2.0, and 1.0-0.6 mya, respectively. The dated fossil record corroborates our paleobiogeographic scenario. We conclude that drastic climatic changes followed by successive temperature oscillations "trapped" most brown frog species in their southern European glacial refugia with enough time to speciate. Substantial dispersal was only possible during extensive interglacial periods of a constant subtropical climate. PMID- 12565040 TI - Phylogenetics of the Tenrecidae (Mammalia): a response to Douady et al., 2002. PMID- 12565043 TI - NMR second moment imaging using Jeener-Broekaert dipolar signals. AB - It is demonstrated that imaging of the 1H NMR second moment can be achieved by using the Jeener-Broekaert (JB) dipolar signal instead of the Zeeman FID signal commonly employed. The JB dipolar signal can be induced by applying a JB pulse sequence, 90 degrees (x)-tau-45 degrees (y)-tau(')-45 degrees (y), which is followed by the time-suspension magic echo sequence, TREV-16TS, for imaging detection. Scanning the imaging detection to cover the whole evolution of the JB dipolar signal finally results in producing spatially resolved JB dipolar signals. The local value of the quantity called the "JB second moment," M(2(JB)), is then estimated from the initial slope of each resolved JB dipolar signal. The M(2(JB)) can be regarded as the "weighted" powder average of the usual second moment. The "weighting" effect due to the JB sequence leads to the tau dependent M(2(JB)) value. The tau dependence is potentially useful for characterizing the second moment distribution resulting from the crystal orientation dependence: For example, in addition to the usual powder average, an approximate distribution range can be deduced by a simple analysis of the tau dependence, serving as a new contrast for materials imaging. This is illustrated by preliminary experiments performed on test samples. PMID- 12565042 TI - Backbone and side chain assignment strategies for multiply labeled membrane peptides and proteins in the solid state. AB - We demonstrate that the SPECIFIC CP technique can be used to obtain heteronuclear correlation (HETCOR) spectra of peptide backbones with greater efficiency than conventional HETCOR methods. We show that similar design principles can be employed to achieve selective homonuclear polarization transfer mediated through dipolar or scalar couplings. Both approaches are demonstrated in a tripeptide with uniform 15N and 13C labeling, and with uniform 15N labeling and natural abundance 13C. In other applications, the high efficiency of the heteronuclear SPECIFIC CP transfer allows discrimination of single amide signals in the 248 residue membrane protein bacteriorhodopsin (bR). In particular, variations are detected in the ordering of the Ala81-Arg82 peptide bond among the photocycle intermediates of bR and SPECIFIC CP is used to correlate 15N and 13C signals from the three Val-Pro peptide bonds. PMID- 12565044 TI - Feasibility of probing boundary morphology of structured materials by 2D NMR q space imaging. AB - It is well known that one-dimensional (1D) q-space imaging allows retrieval of structural information at cellular resolution. Here we demonstrate by simulation that boundary morphology of structured materials can be derived from 2D q-space mapping. Based on a finite-difference model for restricted diffusion, 2D q-space maps obtained from water diffusion inside apertures at various levels of asperity were simulated. The results indicate that the observed ring patterns (diffraction minima) reveal the boundary profiles of the apertures but become blurred in the case of significant variation in aperture size. For uniform size distribution of apertures, a quantitative measure of surface roughness can be established by means of spatial autocorrelation analysis. The results suggest that 2D q-space imaging may allow probing of the boundary morphology of structured materials and possibly biological cells. PMID- 12565045 TI - Analysis of the effect of foreign gases in the production of hyperpolarized 129Xe gas on a simple system working under atmospheric pressure. AB - Experimental conditions that affect the degree of polarization of 129Xe gas were tested for a higher degree of polarization to facilitate a laboratory use of 129Xe NMR, primarily on the effect of addition of foreign gases. When He, N(2), or D(2) gas was added separately to pure Xe gas with natural isotope abundance, D(2) gas gave better results than the others in enhancing the degree of polarization in 129Xe atom. When these gases were added in mixture, however, N(2) plus He was proved to be more efficient than D(2) or He in enhancing the degree of polarization. As a result, the degree of polarization was found to be increased by more than an order, when diluent gases were properly mixed; polarization as high as 35% was reached at gas composition of 5% Xe, 10% N(2), and 85% He, whereas only a few percent was attainable when Xe gas was polarized without mixing any foreign gases [J. Magn. Reson. 150 (2), 156-160 (2001)]. These results were discussed on a basis of quenching and buffer effects of foreign gases. Polarization was also measured after separating the pure Xe gas from the mixture; value of 22% was obtained for the Xe gas isolated after solidification in liquid nitrogen trap. Build-up time of the polarization was also tested, which did not change remarkably depending on the gas composition. PMID- 12565046 TI - Structural fitting of PISEMA spectra of aligned proteins. AB - An algorithm for fitting protein structures to PISEMA spectra is described, and its application to helical proteins in aligned samples is demonstrated using both simulated and experimental results. The formulation of the algorithm in terms of rotation operators yields compact recursion relations that provide a fast and effective way of obtaining peptide plane orientations from chemical and torsion angle constraints. The algorithm in combination with experimental solid-state NMR data results in a method for determining the backbone structures of proteins, since it yields the orientation of a helix as a whole, including its tilt and twist angles, and describes kinks and curves with atomic resolution. Although the algorithm can be applied in an "assignment-free" manner to spectra of uniformly labeled proteins, the precision of the structural fitting is improved by the addition of assignment information, for example the identification of resonances by residue type from spectra of selectively labeled proteins. PMID- 12565047 TI - Correlation of fast and slow chemical shift spinning sideband patterns under fast magic-angle spinning. AB - A new two-dimensional solid-state NMR experiment, which correlates slow and fast chemical shift anisotropy sideband patterns is proposed. The experiment, dubbed ROSES, is performed under fast magic-angle spinning and leads to an isotropic spectrum in the directly detected omega(2) dimension. In the evolution dimension omega(1), the isotropic chemical shift is reduced by a factor S, and spinning sidebands are observed spaced by a scaled effective spinning speed omega(R)/S. These spinning sidebands patterns are not identical to those observed with standard slow magic-angle spinning experiments. Chemical shift anisotropy parameters can be accurately extracted with standard methods from these spinning sideband patterns. The experiment is demonstrated with carbon-13 experiments on powdered samples of a dipeptide and a cyclic undecapeptide, cyclosporin-A. PMID- 12565048 TI - The elimination of magnetic susceptibility artifacts in the micro-image of liquid solid interfaces: internal gradient modulation by the CPMG RF train. AB - Distortions of magnetic resonance images near solid-liquid interface appear as the result of the restriction to spin self-diffusion in the proximity of impermeable boundary as well as of a susceptibility difference. The spectral analysis of spin echo enables to resolve, in a simple way, how various RF gradient pulse sequences reduce the effect of the internal magnetic field induced by the susceptibility difference at interfaces. The 1D diffusion-weighted imaging of water in the narrow notch tested efficiency of some sequence. The notch was milled in a piece of Plexiglas. The method can be used to distinguish the susceptibility effect from the effects of applied gradients when investigating the transport of fluid through a porous structure. PMID- 12565049 TI - Multiplex phase cycling. AB - We discuss a new class of phase cycling procedures, in which a set of individual phase-shifted transients are stored separately in the computer and processed afterwards to yield the separated NMR signals from two or more coherence transfer pathways. In the case of two-dimensional double-quantum spectroscopy, this multiplex acquisition procedure allows the acquisition of pure-absorption spectra in only 62.5% of the time needed by previous methods. PMID- 12565050 TI - Calculation of coherence pathway selection and cogwheel cycles. AB - The selection of proper coherence pathways is a very important aspect of the design of NMR pulse sequences. This article describes a C++ program for the calculation of coherence pathway selection via phase cycles, including a module to calculate cogwheel cycles. Cogwheel phase cycles shorter than the original ones [M.H. Levitt et al., J. Magn. Reson. 155 (2002) 300] are derived and experimentally tested for the MQMAS experiment for 3/2 spins. Some other cogwheel cycles are derived for the MQNQMAS, the STMAS experiment, and a PFG diffusion pulse sequence. This program is publicly available through our website http://www.nyu.edu/projects/jerschow with additional documentation and examples. PMID- 12565051 TI - The Xplor-NIH NMR molecular structure determination package. AB - We announce the availability of the Xplor-NIH software package for NMR biomolecular structure determination. This package consists of the pre-existing XPLOR program, along with many NMR-specific extensions developed at the NIH. In addition to many features which have been developed over the last 20 years, the Xplor-NIH package contains an interface with a new programmatic framework written in C++. This interface currently supports the general purpose scripting languages Python and TCL, enabling rapid development of new tools, such as new potential energy terms and new optimization methods. Support for these scripting languages also facilitates interaction with existing external programs for structure analysis, structure manipulation, visualization, and spectral analysis. PMID- 12565052 TI - Inversion of self-diffusion anisotropy by chemical substitution in smectic A liquid crystals as evidenced by PGSE experiment combined with a quadrupole-coil rotation. AB - A technique of pulsed-field-gradient spin-echo (PGSE) NMR combined with a quadrupole-coil rotation was applied to the study of anisotropic self-diffusion in smectic A liquid crystals. Diffusion anisotropy was found to be inverted by chemical substitution of the terminal groups in homologous compounds: Namely, the diffusion across the layer is faster for the cyano compound, whereas the diffusion within the layer is faster for the trifluoromethoxy compound. PMID- 12565053 TI - 1H detected 1H,15N correlation spectroscopy in rotating solids. AB - We describe new correlation experiments suitable for determining long-range 1H-1H distances in 2H,15N-labeled peptides and proteins. The approach uses perdeuteration together with back substitution of exchangeable protons during sample preparation as a means of attenuating the strong 1H-1H dipolar couplings that broaden 1H magic angle spinning (MAS) spectra of solids. In the approach described here, we retain 100% of the 1H sensitivity by labeling and detecting all exchangeable sites. This is in contrast to homonuclear multiple pulse decoupling sequences that are applied during detection and that compromise sensitivity because of the requirement of sampling between pulses. As a result 1H detection provides a gain in sensitivity of >5 compared to the 15N detected version of the experiment (at a MAS frequency of 13.5kHz). The pulse schemes make use of the favorable dispersion of the amide 15Ns resonances in the protein backbone. The experiments are demonstrated on a sample of the uniformly 2H,15N labeled dipeptide N-Ac-Val-Leu-OH and are analogous to the solution-state suite of HSQC-NOESY experiments. In this compound the 1H amide linewidths at 750MHz vary from approximately 0.67 ppm at omega(r)/2pi approximately 5kHz to approximately 0.20 ppm at omega(r)/2pi approximately 30kHz, indicating that useful resolution is available in the 1H spectrum via this approach. Since the experiments circumvent the problem of dipolar truncation in the 1H-1H spin system, they should make it possible to measure long-range distances in a uniformly labeled environment. Thus, we expect the experiments to be useful in constraining the global fold of a protein. PMID- 12565054 TI - A step closer in the quest for reliable quantification in echocardiography. PMID- 12565055 TI - The right ventricular infundibulum: has it a role? PMID- 12565056 TI - Intracardiac echocardiography guiding successful ablation. PMID- 12565057 TI - Colour encoded endocardial tracking: the current state of play. PMID- 12565058 TI - Visualisation of intra-cardiac structures and radiofrequency lesions using intracardiac echocardiography. AB - AIMS: Fluoroscopy does not allow identification specific anatomical landmarks during electrophysiological studies. Intra-cardiac echocardiography permits visualization of these structures with excellent accuracy, but the optimal method has not been fully described. The aim of this study was to assess the capability of intra-cardiac echocardiography for the visualization of such structures using two different approaches. We also assessed its capability for the evaluation of radio frequency lesions 20 min after catheter ablation of the cavo-tricuspid isthmus. METHODS: Intra-cardiac echocardiography was performed using a 9 MHz rotating transducer in eight consecutive patients (age range: 37-76 years) after radio frequency ablation of the cavo-tricuspid isthmus. The ultrasound catheter was inserted through the femoral vein into the superior vena cava and was pulled back to the inferior vena cava. The echo catheter was then reinserted through the subclavian vein and advanced into the right ventricular apex and was pulled back from the right ventricular to the superior vena cava. Qualitative evaluation and intra-cardiac measurements were performed off-line. RESULTS: The fossa ovalis, the tricuspid valve, and the terminal crest were visible in all patients regardless of the method of introduction of the echo catheter. Left-sided structures were less accurately seen by intra-cardiac echocardiography. The horizontal diameter of the fossa ovalis was 8.9+/-1.8mm. The cavo-tricuspid isthmus was visible using the femoral approach in three patients. The isthmus could be visualized in all patients, and in three patients together with the ostium of the coronary sinus, using the subclavian approach. radio frequency lesions were not visible 20 min after ablation. Additionally, both the left and right ventricles could be seen using the subclavian approach. CONCLUSIONS: The subclavian approach is feasible, safe and superior to visualize the isthmus. Twenty minutes after radio frequency ablation of the cavo-tricuspid isthmus radio frequency lesions are not visible using intra-cardiac echocardiography. PMID- 12565059 TI - The potential value of ultrasonic deformation measurement in differentiating regional ischaemic substrates during dobutamine stress echocardiography. AB - The differentiation between differing regional ischaemic substrates is crucial for decision-making in patients with coronary artery disease. This study demonstrates that quantification of dobutamine stress echocardiography using ultrasonic strain measurement has the potential to identify three differing regional ischaemic substrates (ischaemic, stunned and scarred) in the same patient. The data were validated by traditional analysis of dobutamine stress echo, coronary angiography and correlative quantitative positron emission tomography information. PMID- 12565060 TI - Right ventricular outflow-tract fractional shortening: an applicable measure of right ventricular systolic function. AB - AIMS: Assessment of right ventricular function is important. However, this is not easy to achieve due to the complex anatomy and geometry of the right ventricle, making the evaluation of its function limited. Therefore, a simple reliable and easy method is needed. This study was performed (1) to evaluate the use of right ventricular outflow tract fractional shortening obtained by M-mode echocardiography as a measure of right ventricular systolic function and (2) to determine the relationship between this parameter and other established measurements of right ventricular function such as long axis excursion. METHODS AND RESULTS: Ninety-two consecutive patients referred for echocardiographic assessment of left and right ventricular function, age mean+/-SD was 68+/-14 years, were investigated. Twenty healthy controls, age 46+/-12 years were also studied. M-mode echocardiography was used to measure right ventricular outflow tract fractional shortening and right ventricular long axis excursion. Doppler echocardiography was used for the estimation of right ventricular-right atrial pressure drop and pulmonary artery acceleration time. Right ventricular outflow tract fractional shortening (P<0.0001), right ventricular long axis excursion (P<0.0001) and pulmonary acceleration time (P<0.0001) were reduced in patients compared to controls. Right ventricular outflow tract fractional shortening correlated with long axis excursion (r=0.66 P<0.0001), pulmonary artery acceleration time (r=0.80 P<0.0001) and right ventricular-right atrial pressure drop (r=-0.53 P<0.0001). Right ventricular long axis excursion correlated with right ventricular-right atrial pressure drop though to a lesser significance (r= 0.27 P <0.001). Furthermore, right ventricular outflow tract fractional shortening was reduced in patients with pulmonary hypertension compared to patients without, this difference was not observed in the right ventricular systolic long axis excursion. CONCLUSION: Right ventricular outflow tract fractional shortening provides a simple and non-invasive measure of right ventricular systolic function. In combination with long axis excursion and Doppler velocities they should provide comprehensive assessment of right ventricular function. PMID- 12565061 TI - Effect of age and sex on left atrial morphology and function. AB - AIMS: Left atrial function is abnormal in a wide range of cardiac diseases. This study was designed to assess the effects of normal ageing and sex on left atrial morphology and function. METHODS AND RESULTS: Echocardiography was performed in 123 subjects (age 57 +/- 19 years, range 22 to 89 years, 59 women) with no evidence of cardiovascular disease. M-mode derived left atrial size, B-mode derived left atrial maximal and minimal volumes, and the volume at onset of atrial systole (P-volume) were measured. Left atrial filling, active and passive emptying volumes and ejections fractions, and expansion index were calculated. Subjects were divided into four groups according to age. Left atrial diameter increased with age, with significantly smaller left atrial size in younger subjects. The oldest subjects had significantly higher (P<0.05) left atrial minimal, maximal and P-volume indices. Filling volume index was highest in the oldest subjects (21.9 +/- 5.6 ml/m(2)). Passive emptying volume index was the lowest in those of middle age (10.5 +/- 2.8 ml/m(2)). Active emptying volume index progressively increased with age (P<0.001). Left atrial expansion index and active emptying fraction were not different between the age groups. There was significant difference in passive emptying fraction (P<0.001) with highest values in the youngest (44.7 +/- 7.3%) and lowest values in the oldest subjects (33.6+/ 5.4%). CONCLUSIONS: Age- and sex-related reference values of echocardiographic indices of left atrial morphology and function are reported. Ageing is associated with left atrial dilatation. Left atrial conduit function deteriorates with age while reservoir and pump function are maintained. Left atrial anteroposterior diameter is smaller in women than in men, but overall left atrial function is not influenced by sex. PMID- 12565062 TI - Feasibility and reproducibility of off-line tissue Doppler measurement of regional myocardial function during dobutamine stress echocardiography. AB - AIMS: Off-line post-processing of colour tissue Doppler from digital loops may allow objective quantification of dobutamine stress echocardiography. We assessed the reproducibility of off-line measurements of regional myocardial velocities. METHODS AND RESULTS: Nine observers analysed 10 studies, each making 2400 observations. Coefficients of variation in basal segments from apical windows, at rest and maximal stress, were 9-14% and 11-18% for peak systolic velocity, 16-18% and 17-19% for time-to-peak systolic velocity, 9-17% and 18-24% for systolic velocity time integral, and 18-23% and 21-27% for systolic acceleration. Coefficients of variation for diastolic velocities in basal segments at rest were 11-40%. Coefficients of variation for peak systolic velocity were 10-24% at rest and 14-28% at peak in mid segments, and 19-53% and 29-69% in apical segments. From parasternal windows coefficients of variation for peak systolic velocity were 14-16% in basal posterior, and 19-29% in mid-anterior segments. High variability makes measurement unreliable in apical and basal anterior septal segments. The feasibility of obtaining traces was tested in 92 subjects, and >90% in all basal and mid segments apart from the anterior septum. CONCLUSION: Quantification of myocardial functional reserve by off-line analysis of colour tissue Doppler acquired during dobutamine stress is feasible and reproducible in 11 segments of the left ventricle. The most reliable measurements are systolic velocities of longitudinal motion in basal segments. PMID- 12565063 TI - Radiofrequency catheter ablation of paroxysmal atrial fibrillation; guidance by intracardiac echocardiography and integration with other imaging techniques. PMID- 12565064 TI - Impact of tissue harmonic imaging in patients with distorted left ventricles: improvement in accuracy and reproducibility of visual, manual and automated echocardiographic assessment of left ventricular ejection fraction. AB - AIMS: The aim of this study was to assess the incremental value of tissue harmonic imaging vs conventional echocardiography for evaluating left ventricular ejection fraction by manual and automated quantitation as well as visual estimation in patients with distorted left ventricles. METHODS AND RESULTS: In 25 patients unselected for image quality and with distorted left ventricles who underwent a nuclear study, digital cineloops of standard apical views were acquired by both tissue harmonic imaging and conventional echocardiography and sent to six observers for analysis of visual and quantitative left ventricular ejection fraction. Tissue harmonic imaging improved both the correlation and agreement of all echo techniques with nuclear measures, compared with conventional echocardiography echo, reducing standard errors (SE) to below 10%: for the visual estimate SE=7.5%, for manual tracing SE=6.3% and for automated tracing SE=8%. Tissue harmonic imaging decreased inter-observer variability compared with conventional echocardiography echo for both visual assessment (12.4% vs 18.4%, P<0.05) and quantitative measures (for manual tracing, 8.2% vs 11.8%, P<0.05; for automated tracing, 7.8% vs 16.8%, P<0.05). CONCLUSIONS: In patients with distorted left ventricles unselected for image quality, tissue harmonic imaging improves accuracy and reproducibility of both visual and quantitative echocardiographic assessment of left ventricular ejection fraction. In particular, it promotes automated quantitation by reducing its high standard error into a clinically reasonable range. PMID- 12565065 TI - Native valve endocarditis with aorta-to-left atrial fistula due to Corynebacterium amycolatum. AB - Infective endocarditis remains a pathology with a high rate of complications and mortality. One of the most dramatic complications is abscess formation. A rare evolution of abscess formation is the development of fistula. We describe an 88 year-old woman with an aortic root abscess and aorta-to-left atrial fistula. To our knowledge this has only been described with streptococcus species as causative micro-organism. In this case the abscess was caused by Corynebacterium amycolatum, which is an infrequently found micro-organism. PMID- 12565066 TI - Reversal of acute pulmonary oedema with beta-blockers in hypertrophic cardiomyopathy. PMID- 12565067 TI - Acute thrombosis of mechanical mitral valve prosthesis: echo-Doppler diagnosis and successful treatment with systemic thrombolysis. AB - The case of a 72-year old patient with acute heart failure due to thrombosis of the mechanical mitral prosthesis is presented. The diagnosis was made by transthoracic echocardiography. The patient refused reoperation, and systemic thrombolysis was administered. After thrombolytic infusion we observe the disappearance of the echocardiographic signs of thrombosis, with concomitant improvement of symptoms. PMID- 12565068 TI - Characterization of piezoelectric materials with large piezoelectric and electromechanical coupling coefficients. AB - The relaxor based ferroelectric (1-x)Pb(Zn(1/3)Nb(2/3))O(3)-xPbTiO(3) and (1 x)Pb(Mg(1/3)Nb(2/3))O(3)-xPbTiO(3) single crystals provided new challenges in property characterization because their extraordinarily large piezoelectric coefficients and electromechanical coupling coefficients. Large errors may occur in some of the derived material constants using conventional characterization techniques. This paper will analyze the inadequacy of the traditional characterization methods and provide some basic guidelines for properly characterizing piezoelectric materials with extremely high piezoelectric and electromechanical coupling coefficients. PMID- 12565069 TI - Single fiber model of particle retention in an acoustically driven porous mesh. AB - A method for the capture of small particles (tens of microns in diameter) from a continuously flowing suspension has recently been reported. This technique relies on a standing acoustic wave resonating in a rectangular chamber filled with a high-porosity mesh. Particles are retained in this chamber via a complex interaction between the acoustic field and the porous mesh. Although the mesh has a pore size two orders of magnitude larger than the particle diameter, collection efficiencies of 90% have been measured. A mathematical model has been developed to understand the experimentally observed phenomena and to be able to predict filtration performance. By examining a small region (a single fiber) of the porous mesh, the model has duplicated several experimental events such as the focusing of particles near an element of the mesh and the levitation of particles within pores. The single-fiber analysis forms the basis of modeling the overall performance of the particle filtration system. PMID- 12565070 TI - Automated quantitative analysis of the shift of frequency spectra generated by attenuated signals from contrast microbubbles. AB - The ultrasound-induced harmonic microbubble response spectrum is known to shift to lower frequencies with increasing tissue attenuation. We hypothesized that this shift could be reproducibly detected in received broadband radiofrequency spectra. We used an automatic Gaussian curve-fitting technique to measure the mean harmonic response generated by three different contrast agents at six incremental levels of attenuation. Analytical curve fitting identified a consistent, reproducible, and statistically significant shift in mean harmonic frequency with increasing attenuation. The presented method could be a step toward attenuation estimation by contrast harmonic imaging; optimization of harmonic signal reception by ultrasound systems; and, ultimately, automatic detection of contrast agents in tissue. PMID- 12565071 TI - Integration of a piezoelectric transformer and an ultrasonic motor. AB - Ultrasonic motors are usually operated at an AC voltage higher than a regular battery. This implies the need of a voltage step-up transformer. In this paper, we report the integration of a piezoelectric transformer (PT) with an ultrasonic motor and a simple drive circuit. The stator of the ultrasonic motor and the PT were operated in the same radial vibration mode. Their dimensions were very close to each other yielding nearly matching resonance frequencies. Consequently, they could be combined together without inductor. The drive circuit was designed by using a 555 timer as an astable multivibrator, and high-speed dual MOSFET drivers as a class D half-bridge switching amplifier. This integrated PT-ultrasonic motor performed reasonably without the use of electromagnetic transformer or inductor. PMID- 12565072 TI - Traveling wave ultrasonic motor: coupling effects in free stator. AB - Generally a stator of traveling wave ultrasonic motor (TWUM) consists of piezoelectric transducers (annular plate or rods) coupled by the way of a metallic ring. These transducers divided into halves are excited independently by two electrical signals with different phases of about 90 degrees. So an elastic traveling wave propagates along the circumference of the ring and a rotor pressed on this vibrating surface is then driven by the stator via contact forces. Many difficulties appear in developing TWUM because the contact between the stator and the rotor via a frictional material is very important. However that may be, the first stage consists in obtaining a vibrating stator with optimum characteristics with two symmetrical phases. The aim of this paper is to discuss some coupling effects in a free stator through an enhanced equivalent circuit model. A simple experimental method based on impedance measurements is performed to estimate the coupling characteristics at a low driving voltage. This paper reports results obtained with the free stator of the well known piezoelectric ultrasonic motor "USR60" by Shinsei Co. Ltd. Since the stator behaves as an elastic body, interactions between the two electrical inputs might be described by the introduction of a coupling oscillator. The comparison of experimental and theoretical results leads to validate the new equivalent circuit of the free stator. The presence of coupling impedance could imply a change of electrical supply condition to optimize the TWUM efficiency. The effects of unbalanced features for each electrical input and the applicability of the proposed model to actual operating condition are discussed in the paper. PMID- 12565073 TI - Microwave excitation of ultrasound in graphite-fiber reinforced composite plates. AB - In this paper is demonstrated the effect of microwave beam polarization on the thermal generation of acoustic waves in continuous fiber-reinforced composite laminates. It is found that beam polarization strongly influences the dielectric interaction that leads to thermal losses, bulk expansion, and acoustic wave generation. The oriented graphite fibers in the composite laminate effectively short the microwave fields and reduce the generation efficiency nearly to zero. Ultrasonic waves at several hundred kHz generated in the composite are detected by air-coupled acoustic transducers located on the opposite side of the plate specimen from the 9.41 GHz incident microwave beam. With some averaging signal-to noise ratios of better than 26 dB are obtained. Applying a conventional model of electromagnetic wave scattering in anisotropic media to this experiment yields good agreement between calculations and measured data. Implications for microwave acoustic testing of graphite-reinforced composites are also discussed. PMID- 12565074 TI - Ultrasonic flowmeters: temperature gradients and transducer geometry effects. AB - Ultrasonic flowmeter performance is addressed for the case of cylindrically shaped flowmeters employing two reciprocal ultrasonic transducers A and B so as to measure time-of-flight differences between signals transmitted from transducer A towards B followed by an equivalent signal transmitted from transducer B towards A. In the case where a liquid flows through the flowmeter's measuring section ("spoolpiece"), the arrival times of the two signals differ by an amount related to the flow passing between the two transducers. Firstly, a detailed study of flow measurement errors with mean flow in the laminar flow regime is carried out as a function of the mode index and the transducer diameter/cylinder diameter ratio in the case where no temperature gradients are present in the flowmeter sensor. It is shown that all modes except the fundamental mode overestimate the mean flow by a factor of 33.33% while excitation of the fundamental mode solely give error-free measurements. The immediate consequences are that the flowmeter error decreases as the transducer diameter/cylinder diameter ratio approaches 1 from 0 reflecting the fact that the excitation level of the fundamental mode increases from almost 0 to 1 as this ratio approaches 1 from 0. Secondly, the effect on flowmeter performance due to flow-induced temperature gradients is examined. It is shown that the presence of temperature gradients leads to flowmeter errors at the higher-flow values even in the case where the fundamental mode is the only mode excited. It is also deduced that flowmeter errors in general depend on the distance between transducers A and B whether temperature gradients exist or not. This conclusion is not reflected in the usual definition of flowmeter errors given by the so-called mode-dependent deviation of measurement introduced in earlier works. PMID- 12565075 TI - Contribution to classification of buried objects based on acoustic impedance matching. AB - Determination of material the buried objects are made of could contribute significantly to their recognition, or classification. This is important in detecting buried antipersonnel landmines within the context of humanitarian demining, as well as in a variety of other applications. In this article the concept has been formulated of the approach to buried object's material determination starting with ultrasonic impulse propagation analysis in a particular testing set configuration. The impulse propagates through a characterized transfer material in such a way that a part of it, a reflected wave, carries the information about the buried object's surface material acoustic impedance. The limit of resolution capability is theoretically analyzed and experimentally evaluated and the influencing factors described. Among these, the contact between clean surfaces of the transfer material and buried object is emphasized. PMID- 12565076 TI - Synthetic aperture focusing for defect reconstruction in anisotropic media. AB - Ultrasonic inspection plays an important role in numerous industrial fields. One of the prominent tasks with respect to quantitative nondestructive evaluation is the determination of location, shape, size and orientation of defects. In this respect, the synthetic aperture focusing technique (SAFT) has been successfully applied to isotropic materials over the years. In anisotropic media, however, its application suffers from several phenomena, which are the direction dependence of the ultrasonic velocities, the beam skewing effect and the modified transducer radiation characteristics. In this article, a SAFT imaging algorithm is presented which fully accounts for the nature of wave radiation and propagation within anisotropic materials. For three-dimensional defect reconstruction, the spatial dependence of the ultrasonic group velocities as well as the radiation characteristics of the transducer are exploited--respective algorithms have been implemented for orthotropic material symmetry. Tests have been performed on unidirectional composite material. PMID- 12565077 TI - An investigation of the dependence of ZnO film on the sensitivity of Love mode sensor in ZnO/quartz structure. AB - Love mode acoustic devices are very promising as biosensors in liquid environments because of their high sensitivity. An experimental study of Love mode sensors based on ZnO/90 degrees rotated ST-cut quartz structure with different sputtering conditions to deposit ZnO films is presented. In order to achieve sensor with higher sensitivity, the effects of sputtering substrate temperatures to deposit ZnO films on the sensitivity of viscosity and conductivity were investigated. Phase velocity, sensitivity and temperature coefficient of frequency (TCF) of Love wave devices have been studied. The Love wave sensor has higher sensitivity as sputtering ZnO films on the unheated substrate than that of on the heated substrate. The maximum sensitivity up to 18.77 x 10(-8) m(2) s kg(-1) of ZnO film with thickness of 1.8 microm for a wavelength of 40 microm is much bigger than SiO(2)/quartz structure. In this research, we report ZnO/90 degrees rotated ST-cut quartz structure of Love wave sensors with high sensitivity of viscosity and conductivity in liquid circumstance and TCF of quartz is compensated by ZnO film. PMID- 12565078 TI - Effects of strontium on the surface acoustic wave properties of Sm-modified PbTiO3 ceramics. AB - The Sm-modified lead titanate ceramics with a composition of (Pb(0.88 x)Sr(x)Sm(0.08))(Ti(0.98)Mn(0.02))O(3); x = 0.05-0.25 were prepared by conventional mixed-oxide method. Surface acoustic wave (SAW) properties, including phase velocity, electromechanical coupling coefficient and temperature coefficient of frequency, were measured. The experiments successfully showed that Sr additive is helpful to obtain higher phase velocity and high electromechanical coupling coefficient. The SAW properties of our samples (V(p),k(2)) are better than some commercially-made PZT and PT samples. PMID- 12565079 TI - Prognostic value of exercise cardiac tomography performed late after percutaneous coronary intervention in symptomatic and symptom-free patients. AB - The long-term prognostic value of single-photon emission computed tomography (SPECT) performed late after percutaneous coronary intervention (PCI) has not been extensively evaluated. Moreover, the role of myocardial ischemia at SPECT in symptom-free patients after PCI is not clear. This study was designed to determine the long-term prognostic value of SPECT in predicting cardiac events after PCI in symptomatic and symptom-free patients. Exercise technetium-99m sestamibi SPECT was performed in 206 patients between 12 and 18 months after PCI. All patients were followed for a mean period of 37 +/- 16 months. Cardiac death, nonfatal myocardial infarction, and late revascularization procedures were considered to be events. Myocardial ischemia at SPECT was detectable in 44 patients. During follow-up, 24 patients experienced events (cardiac death in 4 patients, myocardial infarction in 10, and late revascularization in 10). At univariate analysis, the summed stress score (p <0.05) and summed difference score (p <0.001) were significant predictors of cardiac events. Event-free survival curves showed a higher event rate in patients with than without ischemia (p <0.001). The occurrence of cardiac events was higher in the presence of ischemia at SPECT in symptomatic and symptom-free patients (both p <0.001). The results of this study demonstrate that the extent and severity of myocardial ischemia at exercise SPECT performed between 12 and 18 months after PCI predicts cardiac events during long-term follow-up in symptomatic and symptom-free patients. PMID- 12565080 TI - Comparison of late outcome in patients with versus without angina pectoris having reversible perfusion abnormalities during dobutamine stress technetium-99m sestamibi single-photon emission computed tomography. AB - The aim of this study was to assess the prognostic significance of reversible perfusion abnormalities in patients without angina during dobutamine stress technetium-99m sestamibi single-photon emission computed tomography (SPECT). The study comprised 224 patients (age 60 +/- 11 years, 144 men) with completely or partially reversible perfusion abnormalities during dobutamine stress sestamibi SPECT. Follow-up end points were hard cardiac events (cardiac death and nonfatal myocardial infarction). Angina occurred in 93 patients (42%) during the dobutamine stress test (symptomatic ischemia group). The 131 patients without dobutamine-induced angina represented the silent ischemia group. There was no significant difference between patients with and without angina with regard to summed stress perfusion score (5.3 +/- 2.5 vs 5.2 +/- 2.2, p = 0.9) or summed ischemic score (3.1 +/- 1.7 vs 3.2 +/- 1.4, p = 0.7). During a median follow-up of 7.2 years, cardiac death occurred in 14 patients (15%) with and in 21 patients (16%) without angina. Nonfatal myocardial infarction occurred in 8 patients (9%) with and in 13 patients (10%) without angina. In a multivariate analysis model of clinical and perfusion data, independent predictors of cardiac events were age (hazard ratio 1.02, confidence intervals [CI] 1.01 to 1.05 per year increment), diabetes mellitus (hazard ratio 1.9, CI 1.2 to 3.4), and ischemic perfusion score (hazard ratio 2.1, CI 1.3 to 3.8). Patients with silent ischemia defined as reversible perfusion abnormalities without associated angina during dobutamine stress sestamibi SPECT imaging had similar incidences of ischemia and similar cardiac event rates compared with patients with symptomatic ischemia. Therefore, the absence of angina in association with reversible perfusion abnormalities should not be interpreted as a sign of a more benign prognosis. PMID- 12565081 TI - Usefulness of coronary flow reserve over regional wall motion when added to dual imaging dipyridamole echocardiography. AB - Vasodilator stress echocardiography allows semi-simultaneous imaging of left anterior descending (LAD) coronary flow and regional wall function. To assess the relative (and additive?) value of regional flow and function for noninvasive identification of angiographically assessed LAD disease in patients with chest pain syndrome, we studied 230 consecutive in-hospital patients (134 men, aged 63.5 +/- 11 years) with chest pain syndrome and normal regional and global left ventricular function. All patients underwent stress echocardiography with dipyridamole (up to 0.84 mg/kg over 10 minutes), including wall motion analysis by 2-dimensional echocardiography and coronary flow reserve (CFR) evaluation of the LAD artery by Doppler, with or without contrast injection. A new regional wall motion abnormality in >or=2 contiguous segments was required for 2 dimensional echocardiographic positivity. CFR was evaluated as the ratio of dipyridamole to peak diastolic coronary blood flow velocity at rest. All patients underwent coronary angiography within 60 days; a quantitatively assessed diameter reduction >50% of the LAD artery was considered significant. Of the 230 patients, 70 had LAD disease. A regional wall motion abnormality in LAD territory was present in 52 patients, and reduced CFR (<1.9) in 62 patients. Sensitivity for detecting LAD disease was 74% for 2-dimensional echocardiography (95% confidence interval [CI] 64% to 84%) and 81% for CFR <1.9 (95% CI 72% to 90%); specificity was 91% (95% CI 87% to 96%) for 2-dimensional echocardiography and 84% for CFR (95% CI 79% to 90%). Accuracy was 86% for 2-dimensional echocardiography (95% CI 82% to 91%) and 83.5% for CFR (95% CI 79% to 88%). When 2-dimensional echocardiography and CFR criteria were considered, sensitivity increased to 93% (95% CI 87% to 99%), with 80.6% specificity (95% CI 74.5% to 86.7%). CFR was assessed during vasodilator stress echocardiography. Its diagnostic accuracy for detecting LAD disease was comparable to regional wall motion abnormalities. However, the data for flow and function can be complementary in terms of predicting underlying angiographic anatomy, because abnormal wall motion can include coronary artery disease, and negative CFR can exclude it. PMID- 12565082 TI - Comparison of effects of nisoldipine-extended release and amlodipine in patients with systemic hypertension and chronic stable angina pectoris. AB - The efficacy and safety of nisoldipine-extended release (ER) and amlodipine were compared in a 6-week multicenter, randomized, double-blind, double-dummy, parallel group, titration-to-effect trial in patients with stage 1 to 2 systemic hypertension (90 to 109 mm Hg diastolic blood pressure [BP]) and chronic stable angina pectoris. After a 3-week placebo run-in period, patients (n = 120) were randomly assigned to active treatment with either nisoldipine-ER (20 to 40 mg) or amlodipine (5 to 10 mg) once daily, titrated as necessary after 2 weeks to achieve diastolic BP <90 mm Hg. After 6 weeks, the mean reduction in systolic/diastolic BP from baseline was 15/13 mm Hg with nisoldipine-ER and 13/11 mm Hg with amlodipine (p = NS/p = NS). Both drugs resulted in similar BP responder rates (diastolic BP <90 mm Hg in 87% of patients who received nisoldipine-ER and 78% of patients on amlodipine, p = NS) and anti-ischemic responder rates (increasing exercise time >20% in 20% and 27%, respectively [p = NS], and increasing exercise time >60 seconds in 32% and 29% of patients, respectively [p = NS]. Also, after 6 weeks of active therapy, there was a similar mean increase in total exercise duration (23 seconds in the nisoldipine-ER group and 21 seconds in the amlodipine group, p = NS). Neither drug increased heart rate and both decreased frequency of anginal episodes. Adverse events were infrequent, and typically were vasodilator-related effects (including headache and peripheral edema) that occurred with somewhat higher incidence in the nisoldipine-ER group. Thus, nisoldipine-ER and amlodipine provided comparable antihypertensive and anti-ischemic efficacy, and both were generally well tolerated. PMID- 12565083 TI - Time-dependent predictors of primary cardiac arrest in patients with acute myocardial infarction. AB - To understand predictors of cardiac arrest early in acute myocardial infarction (AMI), for the Thrombolytic Predictive Instrument, we developed a multivariable regression model predicting primary cardiac arrest using time-dependent variables based on a case-control study of emergency department (ED) patients with AMI: 65 cases with sudden cardiac arrest and 258 without cardiac arrest. Within the first hour of AMI symptom onset, adjusting for age, systolic blood pressure, serum potassium, and infarct size, increased risk of cardiac arrest was associated with electrocardiographic prolonged QTc interval and a greater sum of ST-segment elevation. After 1 hour, the effect of ST-segment elevation was much reduced and the effect of the QTc interval was reversed, so prolonged QTc appeared protective. Accordingly, for patients presenting 30 minutes after chest pain onset, compared with a QTc of 0.44, the risk for cardiac arrest for patients with QTc of 0.50 was more than doubled (odds ratio [OR] 2.20, 95% confidence intervals [CI] 1.17 to 4.13), whereas for those presenting after an hour, it was much lower (e.g., at 1.5 hours, OR 0.21, 95% CI 0.06 to 0.73). Patients presenting 30 minutes after chest pain onset with a sum of ST elevation of 20 mm had a threefold higher risk than patients with a sum of ST elevation of 5 mm (OR 3.37, 95% CI 1.83 to 6.20). However, if presenting 1.5 hours after chest pain onset, the risk was barely elevated (OR 1.18; 95% CI 1.09 to 1.29). Thrombolytic therapy was protective, halving the odds of cardiac arrest (OR 0.51, 95% CI 0.27 to 0.93). Thus, the relation of prolonged QTc interval and substantial ST segment elevation to cardiac arrest in AMI may be obscured because patients with these risks are more likely to die soon after AMI onset, before ED presentation, and are thereby unavailable for study. Those with prolonged QTc or substantial ST elevation who survive the initial 1.5-hour period are those less susceptible to these risks. PMID- 12565084 TI - Involvement of C-reactive protein obtained by directional coronary atherectomy in plaque instability and developing restenosis in patients with stable or unstable angina pectoris. AB - We investigated whether positive immunohistochemical staining of C-reactive protein (CRP) in initial culprit lesions is related to coronary plaque instability and whether it could affect the outcome of directional coronary atherectomy (DCA). The plasma level of CRP is a reliable marker of the risk of coronary events and restenosis after percutaneous coronary intervention. However, the influence of tissue CRP in atheromatous plaque on plaque vulnerability and restenosis remains unknown. Samples of DCA obtained from 12 patients with stable angina pectoris and 15 patients with unstable angina pectoris were immunohistochemically stained with a monoclonal antibody against CRP. We performed follow-up coronary angiography on 22 of 27 patients to evaluate the presence of restenosis after DCA. Immunoreactivity to CRP was localized to macrophages, smooth muscle cells, and necrotic areas. The ratio of CRP positive cells to total cells was significantly higher in DCA samples from patients with unstable (17.9 +/- 2.0%) than with stable angina (11.0 +/- 2.5%) (p <0.05). Follow-up coronary angiography showed that 12 of 22 patients developed restenosis after DCA. The ratio was also significantly higher in DCA specimens from patients with restenosis (19.3 +/- 2.8%) compared with those without restenosis (11.0 +/- 2.0%) (p <0.05). In addition, the ratio significantly correlated with late luminal loss (r = 0.428, p <0.05) and loss index (r = 0.636, p = 0.0011) after DCA. Immunoreactivity to CRP in coronary atheromatous plaque increases in culprit lesions of unstable angina, and it affects restenosis after DCA. These findings suggest that CRP in atheromatous plaque plays an important role in the pathogenesis of unstable angina and restenosis after coronary intervention. PMID- 12565085 TI - Relative contributions of intimal hyperplasia and vascular remodeling in early cardiac transplant-mediated coronary artery disease. AB - The relative contribution of intimal hyperplasia and vascular remodeling in early transplant coronary artery disease (TxCAD) is unknown. This study was designed to determine the contributions of vascular remodeling and intimal hyperplasia in the initial year after transplantation by intravascular ultrasound (IVUS). Twenty five patients underwent baseline (<6 weeks after transplant) and 1-year angiography and IVUS to evaluate total vessel, luminal, and intimal + medial areas in >or=3 segments of the coronary artery. Nine patients had donor atherosclerotic disease on baseline study (23% of segments), and at 1-year, 21 patients (84%) had intimal hyperplasia (70% of segments). Fourteen patients had positive remodeling in all arterial segments, whereas the remaining 11 had positive and negative remodeling in the same vessel. Mean plaque area and total vessel area increased significantly (p = 0.0001) in proximal, mid, and distal segments, whereas total vessel area was most pronounced in distal segments. Luminal area did not change over time. Of the 87 segments evaluated, 68 (78%) had an increase in total vessel area, 57 (66%) had intimal growth, and 54 (62%) had an increase in luminal area. Although changes in total vessel and luminal area were closely correlated, a decrease in luminal area was associated with positive and negative remodeling. In conclusion, luminal area is generally maintained during the initial transplant year despite significant intimal hyperplasia due to positive remodeling. Reduction in the luminal area results from either inadequate positive remodeling or negative remodeling without intimal growth and often occurs in the same artery. PMID- 12565086 TI - Functional relevance of aldosterone for the determination of left ventricular mass. AB - In experimental studies, the importance of aldosterone for the development of left ventricular (LV) hypertrophy has been demonstrated. In 120 healthy Caucasian men (aged 25 +/- 3 years; blood pressure, 134 +/- 15/86 +/- 12 mm Hg), we determined LV mass (2-dimensionally guided M-mode echocardiography), urinary aldosterone concentration, and the response of aldosterone to angiotensin II infusion (3.0 ng/kg/min). Seventy-six volunteers took part in a follow-up visit after 2 years when urinary aldosterone concentration and LV mass were determined again. At follow-up, LV mass increased in 42 subjects (by 33 +/- 26 g), whereas in 34 subjects LV mass decreased (by 27 +/- 22 g). Between the 2 groups, only the change in urinary aldosterone concentration over time was significantly different (group with increased LV mass had an increase in urinary aldosterone concentration by 2.5 +/- 5.4 microg/day; group with decreased LV mass had a decrease in urinary aldosterone concentration by 0.7 +/- 4.6; p <0.01 between groups). In accordance, we found significant correlations between changes in LV mass and changes in urinary aldosterone concentration (r = 0.29, p <0.05) and between changes in LV mass and the response of aldosterone to angiotensin II at baseline (r = 0.25, p <0.05). Both changes in aldosterone concentration over time and the response of aldosterone to angiotensin II were related to changes in LV mass over time. These data underscore the importance of aldosterone for the development of LV hypertrophy. This process is already evident in young subjects with apparently small changes in LV mass over a mean follow-up period of 2 years. PMID- 12565087 TI - Leslie David Hillis, MD: a conversation with the editor. [interview by William Clifford Roberts]. PMID- 12565088 TI - Long-term changes in exercise capacity, quality of life, body anthropometry, and lipid profiles after a cardiac rehabilitation program in obese patients with coronary heart disease. PMID- 12565089 TI - Impairment of myocardial perfusion in both culprit and nonculprit arteries in acute myocardial infarction: a LIMIT AMI substudy. PMID- 12565090 TI - Echocardiographic characteristics of patients surviving nonsurgically treated left ventricular pseudoaneurysm after acute myocardial infarction. PMID- 12565091 TI - Relation of C-reactive protein and interleukin-6 to culprit coronary artery plaque size in patients with acute myocardial infarction. PMID- 12565092 TI - Increased concentrations of tirofiban in blood and their correlation with inhibition of platelet aggregation after greater bolus doses of tirofiban. PMID- 12565093 TI - Comparison of clinical, angiographic, and intravascular ultrasound parameters after direct stenting versus predilation. PMID- 12565094 TI - Impact of acute myocardial infarct location on myocardial salvage after stenting or thrombolysis (results from the STOPAMI 1 and 2 trials): Stent versus Thrombolysis for Occluded coronary arteries in Patients with Acute Myocardial Infarction. PMID- 12565095 TI - Rapid sealing of coronary perforations using polytetrafluoroethylene-covered stents. PMID- 12565096 TI - Effects of tirofiban on hemostatic activation and inflammatory response during cardiopulmonary bypass. PMID- 12565097 TI - Comparison of effectiveness of statin monotherapy versus statin and niacin combination therapy in primary prevention and effects on calcified plaque burden. PMID- 12565098 TI - Comparison of left atrial volume assessed by magnetic endocardial catheter mapping versus transthoracic echocardiography. PMID- 12565099 TI - Peripheral conduction disease in left ventricular dysfunction. PMID- 12565100 TI - Dissociation between exercise hemodynamics and exercise capacity in patients with chronic heart failure and marked increase in ejection fraction after treatment with beta-adrenergic receptor antagonists. PMID- 12565101 TI - Prevalence and severity of mitral regurgitation in chronic systolic heart failure. PMID- 12565102 TI - Effect of angiotensin-converting inhibitor or angiotensin receptor blocker on one year survival in patients >or=65 years hospitalized with a left ventricular ejection fraction >or=50%. PMID- 12565103 TI - Usefulness of magnetic resonance imaging in diagnosis of arrhythmogenic right ventricular dysplasia and agreement with electrocardiographic criteria. PMID- 12565104 TI - Association of leisure-time physical activity on inflammation markers (C-reactive protein, white cell blood count, serum amyloid A, and fibrinogen) in healthy subjects (from the ATTICA study). PMID- 12565105 TI - Effect of hormone replacement therapy on tissue factor activity, C-reactive protein, and the tissue factor pathway inhibitor. PMID- 12565106 TI - Myocardial perfusion in children with sickle cell disease. PMID- 12565107 TI - Noninvasive visualization and measurement of posterior descending coronary artery flow velocity by transthoracic Doppler echocardiography in normal children. PMID- 12565108 TI - Definitive repair of double-outlet right ventricle with subaortic ventricular septal defect and pulmonary atresia in adulthood. PMID- 12565109 TI - William Dock's contribution to cardiac auscultation. PMID- 12565110 TI - Statin and adverse events: it does not add up. PMID- 12565112 TI - FDA use of the black box warning: time for reevaluation as a safety tool. PMID- 12565113 TI - Anesthesia care for living-related liver transplantation for infants and children with end-stage liver disease: report of our initial experience. AB - STUDY OBJECTIVE: To describe our initial experience of the perioperative anesthetic care provided to pediatric recipients during living-related liver transplantation. DESIGN: Cohort review of the perioperative anesthetic care for living-related liver transplantation. SETTING: Tertiary referral and postgraduate teaching hospital. PATIENTS: 27 children (20 males, 7 females) with end-stage hereditary metabolic liver disease requiring living-related liver transplantation. INTERVENTION: Perioperative care was administered during living related liver transplantation. MEASUREMENTS: The major intraoperative physiologic events and concerns are described, as well as the anesthesia technique for pediatric living-related liver transplantation anesthesia. Intraoperative changes in physiologic parameters and the intraoperative requirements in our series are also reported. MAIN RESULTS: During a 30-month period, 27 children (20 males and 7 females) were scheduled for transplantation with an hepatic graft from a living related donor. Twenty-six children received a graft from a living-related donor, and one was retransplanted with a cadaveric graft because of graft failure, and one child received a cadaveric graft because of the lack of a suitable donor. All patients received intravenous (IV) anesthesia with fentanyl, midazolam, and cisatracurium, and were ventilated with oxygen/air. Mean induction and presurgical preparation time was 1.18 hours, with a surgical time of 6.55 hours. All but one patient was extubated on the evening of the operating day after receiving a mean dose of 8.67 microg kg(-1) hr(-1) of fentanyl and a mean dose of 0.124 mg kg(-1) hr(-1) midazolam. The need for crystalloid infusion was 24.0 mL kg(-1) hr(-1), fresh frozen plasma (FFP)16.63 mL kg(-1) hr(-1), and red blood cells 7.98 mL kg(-1) hr(-1). There was no mortality and no anesthetic-related morbidity in our series. CONCLUSIONS: Total IV anesthesia with fentanyl, midazolam, and cisatracurium, after preoperative optimization, is a well tolerated approach for children undergoing living-related liver transplantation and offers quick recovery. This anesthetic technique was aimed at minimizing the effects on the cardiovascular system, and also any consequences related to the possible occurrence of a reperfusion syndrome. Fluid balance was aimed at optimizing flow through the hepatic graft and preventing thrombosis of vascular anastomoses. PMID- 12565115 TI - Pilot study on the effect of tourniquet use on sufentanil pharmacokinetics. AB - STUDY OBJECTIVE: To test our hypothesis that sequestration of sufentanil can occur during surgery when a pneumatic tourniquet is used. DESIGN: Prospective, randomized study. SETTING: Operating room and recovery room of a university hospital. PATIENTS: 16 ASA physical status I and II patients scheduled for orthopedic surgery with pneumatic tourniquet use. INTERVENTION: Patients were randomized to three groups. Sufentanil was given intravenously at 0.5 microg kg( 1) bolus at the same time that a constant infusion was started at 0.5 microg kg h(-1). In Group 1, continuous infusion of sufentanil was stopped when the tourniquet was released (n = 6). In Group 2, continuous infusion of sufentanil was stopped 15 minutes after tourniquet release (n = 6). In Group 3, as a control group, the sufentanil bolus was started after tourniquet inflation (n = 4). MEASUREMENTS: Plasma sufentanil concentrations were determined by radioimmunoassay. To compare pharmacokinetic results, a simulation of the sufentanil plasma concentrations was achieved. MAIN RESULTS: Exsanguination and inflation of the pneumatic tourniquet had no significant effect on pharmacokinetic results. In 75% of patients, a significant increase in sufentanil plasma concentration occurred between 30 and 60 minutes after tourniquet deflation in all three groups, probably as a result of patient mobilization. One respiratory distress event occurred in a Group 2 patient following extubation at 55 minutes after the end of the sufentanil infusion. The rebound of sufentanil concentration was higher in Group 2; it may be due to a reduced effect of the restoring circulation in the ischemic leg by a prolonged infusion after tourniquet deflation. CONCLUSIONS: Using a pneumatic tourniquet induces transient changes in the pharmacokinetics of sufentanil. These changes may have clinical relevance during the first hour after tourniquet release. PMID- 12565114 TI - Comparison of ropivacaine-fentanyl patient-controlled epidural analgesia with morphine intravenous patient-controlled analgesia for perioperative analgesia and recovery after open colon surgery. AB - STUDY OBJECTIVE: To compare the effects of ropivacaine-fentanyl patient controlled epidural analgesia (PCEA) with morphine intravenous (IV) patient controlled analgesia (PCA). DESIGN: Prospective, randomized, multicenter trial. SETTING: Five university-affiliated hospitals. PATIENTS: 41 patients undergoing colon surgery. INTERVENTION: Patients were randomized to receive either standardized combined epidural/general anesthesia followed by PCEA with ropivacaine 0.2% and fentanyl (2 microg/mL) or standardized general anesthesia followed by morphine IV PCA. All patients participated in a standardized postoperative clinical pathway. MEASUREMENTS AND MAIN RESULTS: Analgesia was assessed with visual analog scale (VAS) scores. Postoperative recovery was assessed by completion of prospectively defined discharge milestones and time until discharge. Statistical analyses included nonparametric and contingency table analyses. The PCEA group had better analgesia (> 50% reduction in pain scores, assessed both at rest and during a cough) for the first 3 days after surgery (p < 0.0,005). The PCEA group achieved discharge milestones approximately 36 hours faster (p < 0.002), but time until discharge was similar between groups. CONCLUSIONS: Ropivacaine-fentanyl PCEA provides superior analgesia, reduced opioid requirement, and more rapid recovery after colon surgery. PMID- 12565116 TI - Training of residents in peripheral nerve blocks during anesthesiology residency. AB - STUDY OBJECTIVE: To survey American anesthesiology residency program directors to determine the availability and extent of training in peripheral nerve block techniques. DESIGN: Survey questionnaire was mailed and faxed to 132 American anesthesiology residency program directors and followed up 4 weeks later with another mailing to nonresponders. SETTING: University medical center. MEASUREMENTS AND MAIN RESULTS: Of the 132 American anesthesiology residency program directors surveyed, 69 (52%) responded. Of the responders, 40 (58%) offered a specific peripheral nerve block rotation. The rotation was of 1 month's duration in 61% of these programs. Formal instruction was administered during the rotation in 69%. The regional instruction approach consisted of a nerve stimulator (98%), paresthesia (75%), and transarterial (85%). Multimedia, mannequins, and cadaver dissection were used infrequently (13-25%). During the rotation, residents performed a variety of blocks, but the number of each block varied from 2 (supraclavicular) to 10 (axillary). These blocks were performed in the operating room in 48% of programs. Finally, in the programs with a specific peripheral nerve block rotation, residents were evaluated. CONCLUSIONS: Specific peripheral nerve block rotations are not always included in anesthesiology residents' curriculum. In addition, residents in programs with a specific nerve block rotation are exposed to a greater number of peripheral nerve block techniques than those who do not have such a rotation included in their curriculum. PMID- 12565117 TI - The use of cardioselective beta-blockers in a patient with idiopathic hypertrophic subaortic stenosis and chronic obstructive pulmonary disease. AB - The beta-adrenergic receptor blocking drugs are commonly used in the treatment of patients with idiopathic hypertropic subaortic stenosis (IHSS). These drugs, however, are contraindicated in patients with chronic obstructive pulmonary disease (COPD). We report the anesthetic management of a patient with IHSS complicated by severe COPD. We concluded that the beta(1) selective, ultra-short acting beta-blocker, esmolol, can be used intraoperatively when both conditions are present. The pathophysiology and the commonly used anesthetic drugs and practices for treatment of patients with IHSS are reviewed. PMID- 12565118 TI - Spinal anesthesia for a parturient with an iatrogenic skull base defect and CSF leak. AB - A patient with an iatrogenic skull base defect and cerebrospinal fluid leak presented with rupture of amniotic membranes at 37 weeks' gestation. She had daily rhinorrhea that began in her first trimester of pregnancy and 11 months after endoscopic sinus surgery performed for chronic sinusitis. Consultation among the otolaryngologist, obstetrician, and obstetrical anesthesiologist were completed during the patient's second trimester to determine the most desirable type of delivery and anesthesia. A cesarean section was performed so as to avoid pushing in the second stage of labor that could worsen the cerebrospinal fluid leak. A spinal anesthetic was performed uneventfully. The patient had an unremarkable recovery from the operative delivery and spinal anesthesia. The pathophysiology, management, and anesthetic concerns of patients with cerebrospinal fluid leaks are reviewed. PMID- 12565119 TI - Withdrawal symptoms in a patient receiving intrathecal morphine via an infusion pump. AB - We describe a case of a failed implanted, programmable, intrathecal infusion pump and its external programmer-leading to morphine withdrawal symptoms, in a patient with failed back syndrome. PMID- 12565120 TI - Droperidol: should the black box be light gray? AB - In December 2001, the United States Food and Drug Administration (FDA) added a "black box" warning to the labeling for droperidol stating that all doses, even those typically used for postoperative nausea and vomiting, were potentially associated with malignant ventricular dysrhythmias, including torsade de pointes. The 19 cases in which droperidol doses less than 10 mg were allegedly associated with such dysrhythmias are reviewed in detail. Confounding issues present in a majority of the cases make it difficult to incriminate droperidol as the likely cause of the reported adverse events. PMID- 12565121 TI - Initial evaluation of a new optic laryngoscope blade. AB - We report on a new optic laryngoscope blade that provides two views of the larynx during tracheal intubation. The availability of an alternative direct view of the larynx may improve the ability of anesthesia providers to observe the tracheal tube passing through the vocal cords when using a Macintosh laryngoscope blade. The optic port improved visualization of passage of the endotracheal tube in obese patients. However, further studies are needed in patients with difficult airways to determine whether this new blade will ease the ability of practitioners to insert a tracheal tube. PMID- 12565122 TI - A modified lateral approach for a single-shot sciatic nerve block in the popliteal fossa. PMID- 12565123 TI - Free, downloadable, peer-reviewed educational resources--an idea whose time has come. PMID- 12565124 TI - Acquisition and maintenance of endoscopic skills: developing an endoscopic dexterity training system for anesthesiologists. AB - The acquisition and maintenance of essential psychomotor skills that are only required sporadically is a significant problem in medical training and practice. It is of particular relevance to anesthesiologists with regard to fiberoptic intubation, a technique that may be under-utilized despite its central role in the management of the difficult airway. Dexterity deficit due to current training models, dexterity decay due to lack of practice, and situational stress related to the clinical environment may combine to impede effective training and confident use of endoscopes in airway management. We describe an educational resource (Dexter), which has been developed to overcome these problems. Dexter is a nonanatomical, endoscopic dexterity training system designed to encourage practice and help establish and maintain a state of procedural readiness, even if clinical exposure to difficult airway situations is sporadic. PMID- 12565125 TI - A new video laryngoscope-an aid to intubation and teaching. AB - In recent decades, video techniques have been employed in the majority of endoscopic procedures because of several distinct advantages provided. These include the following: The displayed anatomy is magnified. Recognition of the anatomical structures and anomalies is easier, and manipulation of airway devices is facilitated. When assistance is required, the operator and assistant can coordinate their movements because each sees exactly the same image on the video monitor. As a result, video techniques have become the method of choice in teaching. The Video Macintosh Intubating Laryngoscope System (VMS) was designed employing a standard Macintosh blade and laryngoscope handle. A camera was incorporated into the handle with a short image and light bundle. The magnified anatomy is displayed on an 8-inch monitor, which is attached to a swivel arm on a small cart. Observation and manipulation can be performed in one axis.A total of 235 patients were studied and were divided into two groups: Group A (n = 217), in whom intubation was thought unlikely to be difficult, and Group B (n = 18), in whom difficulty with intubation was anticipated. External laryngeal manipulation (ELM) was required in 22 of the 217 Group A patients (10%). All intubations but one in this group were successful. In the second group (B) of 18 patients who had anatomical conditions that suggested that direct laryngoscopy might be challenging, all 18 cases required ELM but all were successfully intubated using the VMS. The improved coordination afforded by an image on a video monitor seen by both the assistant providing laryngeal manipulation and the anesthesiologist handling the laryngoscope results in a significant advantage over the conventional laryngoscope technique. As a consequence, the learning curve is short. In our view, video laryngoscopy will become the method of choice in teaching. PMID- 12565126 TI - Serotonin 1B knockout mice exhibit a task-dependent selective learning facilitation. AB - Serotonin 1B knockout (KO) mice exhibit better spatial reference memory performance in the Morris water maze than their wild type (WT) controls. The present study was aimed at dissecting the underlying cognitive bases of this facilitation using a stepwise water maze paradigm. The performance of KO mice did not differ from WT in a single start-goal task, nor when using two opposite starts. However, KO mice exhibited better performance in stages requiring cognitive flexibility or the higher level of spatial navigation planning (standard version). In a short-term memory version of the task, no such genotype effect was observed, confirming our previous findings. These results suggest that the serotonin 1B receptor gene deletion selectively enhances learning performance when the cognitive requirement of the task is elevated. PMID- 12565127 TI - Importance of IgG2c isotype in the immune response to beta-amyloid in amyloid precursor protein/transgenic mice. AB - A careful analysis of the immune response to immunization of amyloid precursor protein/transgenic (APP/Tg) mice with beta-amyloid (Abeta) may provide insights into why a subset of the patients in a clinical trial receiving Abeta immunotherapy developed encephalomyelitis. Characterization of isotypic immune responses have been reported in different APP/Tg models. In these studies the relative ratios of IgG1 to IgG2a anti-Abeta antibodies has been used as an indirect measure of T helper 1 (Th1) and Th2 types immune responses. However, it has previously been shown that certain strains of mice, C57Bl/6, C57Bl/10, SJL, and NOD, have an IgG2c rather than an IgG2a gene. Since a substantial number of Abeta-immunization studies rely on APP/Tg mice that have at least one parental C57Bl/6 strain, we have investigated whether antibodies specific for IgG2a can be used for characterization of antibody isotypes in APP/Tg2576 mice. Our results suggest that APP/Tg2576 and major histocompatibilty complex-matched parental strains are not expressing IgG2a, producing instead IgG2c anti-Abeta antibodies. PMID- 12565128 TI - Pre and postsynaptic effects of metabotropic glutamate receptor activation on neonatal rat hypoglossal motoneurons. AB - Activation of metabotropic glutamate receptors by (+/-)-1-aminocyclopentane-trans 1,3-dicarboxylic acid (t-ACPD) produced pre- and postsynaptic effects on hypoglossal motoneurons (HMs) patch clamped in thin slices from neonatal rat brainstem. t-ACPD depolarized HMs (with parallel increase in R(in)), induced membrane oscillations, and depressed glutamatergic transmission. However, t-ACPD neither changed the discharge pattern of HMs induced by current pulses nor modulated the spike afterhyperpolarization. The only apparent alteration in firing was the delayed onset of the second spike in a train. The present results provide new evidence that activation of metabotropic glutamate receptors could influence HM excitability by changing the cell responsiveness to synaptic inputs rather than via obvious alterations in the spike firing properties. PMID- 12565129 TI - Correlation of oxidative stress and the loss of the nicotinic receptor alpha 4 subunit in the temporal cortex of patients with Alzheimer's disease. AB - The correlation between oxidative stress and the loss of nicotinic acetylcholine receptor (nAChR) alpha4 subunit has been investigated in the temporal cortex from patients with Alzheimer's disease (AD). The level of lipid peroxidation was significantly increased in AD brains, whereas there were no significant changes in protein oxidation either in whole tissues or the cellular membrane protein parts between AD brains and controls. The nAChR alpha4 subunit at protein level was significantly decreased in AD brains. Furthermore, there was a correlation between the increased levels of lipid peroxidation and the decreased numbers of the alpha4 subunit protein in AD brains. We suggest that lipid peroxidation might be a sensitive target in AD brain and related the mechanism of the loss of the nAChR alpha4 subunit in the disease. PMID- 12565130 TI - Antiserum against human glucose transporter 5 is highly specific for microglia among cells of the mononuclear phagocyte system. AB - Human monocytes and a variety of tissue macrophages, including microglia, were studied immunohistochemically to determine the expression of a novel microglial marker, human glucose transporter 5 (hGLUT5), in these cells. The hGLUT5 was not expressed in most peripheral macrophages in the normal state, but weakly expressed in some foamy macrophages in atherosclerotic lesions. There was no hGLUT5 reactivity in blood monocytes. In the lesions of brain infarcts, foamy macrophages (predominantly monocyte-derived cells) in the ischemic core were mostly negative for hGLUT5, while activated and phagocytic microglia in the transitional zone were consistently positive. The present study indicated that unlike other microglial markers, hGLUT5 is rarely present in peripheral macrophages, and that hGLUT5 immunohistochemistry is useful in distinguishing microglia-derived macrophages from monocyte-derived macrophages in acute necrotic lesions. PMID- 12565131 TI - No association of mutations and mRNA expression of WFS1/wolframin with bipolar disorder in humans. AB - Association of WFS1 (wolframin) and bipolar disorder has been suggested by psychiatric manifestations in patients or non-symptomatic carriers of Wolfram disease and linkage of bipolar disorder with 4p16, the locus of WFS1. Five studies of WFS1 in bipolar disorder did not support this association, although possible association of several missense mutations has not been excluded yet. In this study, four such mutations were genotyped in 184 patients with bipolar disorder and 207 controls. None had the A559T and A602V mutations, and no association of G576S and H611R with bipolar disorder was found. We also quantified the expression levels of WFS1 mRNA in the postmortem brains of patients with bipolar disorder, depression, schizophrenia, and controls. There was no significant difference of the expression levels. These results did not support the pathophysiological significance of WFS1 in bipolar disorder. PMID- 12565132 TI - Adenosine triphosphate as a stimulant for nociceptive and non-nociceptive muscle group IV receptors in the rat. AB - For cutaneous and articular nociceptors, adenosine triphosphate (ATP) has been shown to be an effective stimulant. The aim of present study was to find out if among muscle afferents ATP-sensitive group IV receptors are present, and if differences in ATP-responsiveness existed between nociceptive and non-nociceptive units. Single fibre activity was recorded from group IV afferents innervating the rat gastrocnemius-soleus muscle. Using mechanical stimuli the fibres were classified as nociceptive or non-nociceptive ones. ATP was injected intramuscularly into the receptive field of the fibre. Results were: (1) ATP at a concentration that is present in muscle cells elicited excitations in 67% of the fibres; (2) ATP excited both nociceptive and non-nociceptive receptors; and (3) the acidity of the ATP solution was not an important factor for the ATP effect. PMID- 12565134 TI - Diurnal changes in corticotropin-releasing hormone messenger RNA in the rat thalamus. AB - Corticotropin-releasing hormone (CRH) is critical for mediating the stress response. CRH messenger RNA (mRNA) is present in a variety of brain regions including the thalamus and thalamic CRH mRNA concentrations increase in response to stress exposure. The present study assessed changes in basal CRH mRNA concentrations in the rat thalamus during different times of the day. Using in situ hybridization, we demonstrated that thalamic CRH mRNA levels exhibited more than two-fold increases during the dark phase between 20:00 and 02:00 h, followed by a decrease at 08:00 and 14:00 h during the light phase. Dramatic changes in thalamic CRH mRNA levels may have important implications for the possible role of thalamic CRH systems in waking, arousal, and the stress response. PMID- 12565133 TI - Temporal regulation of CD81 following retinal injury in the rat. AB - Following retinal injury, glial cells within the retina undergo a response that is characterized by the proliferation of astrocytes, Muller cells, and retinal pigment epithelial cells. CD81, a small membrane protein known to be involved in cell proliferation, is up-regulated after injury. This study focuses on the temporal regulation of CD81, relating the expression of this protein to glial fibrillary acidic protein (GFAP), the classic marker of gliosis. CD81 levels were elevated at 7 days after injury and remained elevated at 30 days after injury; GFAP was increased at 7 days and continued to increase until 30 days post injury. This association of CD81 with glial reactivity may provide a clue to the regulation of the proliferative response following retinal injury. PMID- 12565135 TI - Activation of the peripheral sympathetic nervous system increased the expression of cyclooxygenase-2 (COX-2) mRNA in mouse calvaria. AB - Pharmacological stimulation of adrenoceptor was demonstrated to increase the synthesis of prostaglandin (PG)-E(2), well known to modulate bone metabolism by regulating development and function of osteoclasts and osteoblasts, in cultured osteoblastic cells. Recently, intracerebroventricular (i.c.v.) injection of lipopolysaccharide (LPS), which caused the inflammatory stimuli in the brain, was demonstrated to increase the outflow of the peripheral sympathetic nervous system. In this study, to clarify the physiological role of sympathetic nerves to bone metabolism in vivo, we examined the effect of LPS (i.c.v.) on the expression of cyclooxygenase-2 (COX-2) mRNA in mouse calvaria, using reverse transcription polymerase chain reaction analysis. The expression of COX-2 mRNA was increased by LPS (i.c.v.) in mouse calvaria. The treatment with the neurotoxin 6 hydroxydopamine or beta-blocker inhibited the central LPS-induced COX-2 mRNA in mouse calvaria. In addition, the treatment of calvaria with isoprenaline, beta agonist, or noradrenaline increased PGE(2) synthesis in the organ culture system. These findings show that central LPS-induced COX-2 mRNA was mediated by the activation of postganglionic sympathetic nerve fibers and beta-adrenoceptor in mouse calvaria and suggest that in vivo activation of the sympathetic nervous system modulates bone metabolism. PMID- 12565136 TI - Human hypnosis: autonomic and electroencephalographic correlates of a guided multimodal cognitive-emotional imagery. AB - The effects of a guided neutral and unpleasant imagery involving several sensory modalities were studied in hypnotized subjects. Heart rate (HR), respiratory frequency (RF), tonic skin resistance and different electroencephalographic rhythms were evaluated during a long-lasting hypnotic session including the guided suggestion of a neutral (NS) and an unpleasant (US) imagery, each preceded by a hypnotic relaxation rest period. During NS, the absence of autonomic changes, associated with electroencephalographic gamma power decrement and theta1 power increment, indicated the prevalence of relaxation on the expected task related modifications. In contrast, US elicited HR and RF increments together with higher electroencephalographic gamma, beta3 and beta2 activities. Thus, hypnotic state appears to prevent the autonomic responses expected during the neutral stimulation, while the emotional valence of the unpleasant imagery overwhelms the hypnosis-related relaxation. PMID- 12565137 TI - Properties of neurons from the rat medial vestibular nucleus in microexplant culture. AB - This study is a first step in an attempt to identify the factors which determine and maintain the electrophysiological phenotype(s) of mature neurons of the medial vestibular nucleus (MVN). We cultured MVN microexplants obtained from slices of the brainstem of newborn rats, using a hollow punching needle. The electrophysiological maturation of the neurons was followed by analyzing their responses to 1 s steps of current of increasing amplitude. The maximal number of spikes that was generated in response to such stimuli increased dramatically over time in vitro. However, even after 28 days in vitro, it did not exceed about 60 spikes/s. At this stage of culture, the input-output properties of the spike generator of the MVN neurons were similar to those observed in brainstem slices of newborn rats, but clearly inferior to those of adult neurons which can generate sustained firing up to 150-200 spikes/s. PMID- 12565138 TI - Induction of inducible nitric oxide synthase expression in activated microglia following domoic acid (DA)-induced neurotoxicity in the rat hippocampus. AB - Neuronal degeneration followed by glial activation (microglia and astrocytes) and nitric oxide synthase (NOS) expression in the hippocampus was investigated at 3 months after domoic acid (DA) administration and compared with DA treated rats at 5 days time interval which was reported earlier. Massive degeneration with complete absence of neurons in the hippocampal CA1 and CA3 regions and hypertrophied microglial cells showing intense immunoreaction with the antibody OX-42 was observed at 3 months after DA administration. Sparsely distributed OX 42 positive microglial cells were observed in the hippocampus of control rats at 3 months after saline treatment No apparent changes could be observed in the immunoreactivity of GFAP at 3 months after saline and DA administration. Neuronal nitric oxide synthase immunoreactive neurons were completely absent in the hippocampus at 3 months after DA administration. In contrast, nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemical analysis revealed absence of NADPH-d reactivity in the neurons, but positive reactivity in the microglial cells of CA1-CA3 regions in the hippocampus after DA treatment. Double immunofluorescense revealed co-expression of inducible nitric oxide synthase with immunoreactive OX-42 positive microglial cells in the hippocampal subfields at 3 months after DA administration. The microglia-produced NO appears to be a secondary phenomenon in the prolonged inflammatory process following DA induced neuronal degeneration. PMID- 12565139 TI - Facilitation of both stretch reflex and corticospinal pathways of the tibialis anterior muscle during standing in humans. AB - Excitability of both stretch reflex (SR) and motor evoked potential (MEP) elicited in the tibialis anterior (TA) muscle by transcranial magnetic stimulation were tested in standing humans. The results demonstrated significantly greater values for both SR and MEP in the TA while standing than while in the supine posture, although background electromyographic activity was silent in the two conditions. Taken together with previous reports that both pathways are facilitated in the TA at the early stance phase of human walking, our findings suggest that a common neural mechanism underlies both observations, one that might be functionally relevant for securing ankle joint stabilization during upright standing. PMID- 12565140 TI - Testosterone regulates androgen and estrogen receptor immunoreactivity in rat substantia nigra pars reticulata. AB - At postnatal day (PN)1, there are sex differences in gonadal receptor expression in the rat substantia nigra pars reticulata (SNR). Male pups have lower levels of androgen receptor (AR) and estrogen receptor (ER)beta immunoreactivity (IR) compared to female pups, while ERalpha IR is equally expressed in the two sexes. To test whether these differences are due to sex differences in testosterone exposure, we injected female pups with testosterone propionate (TP) on the day of birth and analyzed the levels of AR and ER IR at PN1. TP-treated females have lower levels of AR and ERbeta IR than control, while there are no differences in the levels of ERalpha IR. TP treatment did not affect the number of AR and ER expressing cells. The regulation of SNR AR and ERbeta IR by testosterone may be important for the development of sex-specific functional systems involved in motor control. PMID- 12565141 TI - Chronic ethanol consumption restores the age-related decrease in neurogranin mRNA level in the hippocampus of mice. AB - Neurogranin (Ng) is a Ca(2+)-sensitive calmodulin-binding neuron-specific protein that has been implicated in the regulation of numerous post-synaptic signalling pathways. Here, we investigate the effects of 5 months low level ethanol consumption (approximately 20% of total calories intake) on Ng mRNA expression in the brain of adult (approximately 7-8 months) and aged (approximately 21-22 months) mice using in situ hybridization histochemistry. Results showed that ageing was accompanied by a decrease in amounts of mRNA coding for Ng, especially in the hippocampus (approximately 25% of adults) known to play a critical role in higher cognitive functions. Chronic ethanol consumption restored this decline up to pre-senescent (adult) levels without altering Ng mRNA levels in adult mice. On the basis of recent data indicating a central role for Ng in the regulation of hippocampal synaptic plasticity and spatial learning, our results suggest that moderate ethanol consumption might have a beneficial influence on cognitive deterioration during senescence. Such a possibility is in fact congruent with recent follow-up studies conducted in elderly people. PMID- 12565142 TI - Cerebral hemodynamics and brain oxygen changes related to gravity-induced loss of consciousness in rhesus monkeys. AB - The aim of the present experiment was to study the influence of +Gz acceleration (head-to-foot inertial forces) onset on cerebral oxygenation changes (cerebral oxy- and deoxy-hemoglobin) and cerebral blood volume (CBV) in order to evaluate the role of cerebral hypoxemia and ischemia in the appearance of +Gz-induced loss of consciousness (G-LOC). We used five rhesus monkeys which were equipped with near infrared spectroscopy optodes fixed onto the parietooccipital cranial bone. G-LOC (isoelectric electrocorticogram) was detected with silver balls electrodes in contact with the dura matter. The animals were centrifuged up to +12 Gz with two onset rates (0.1 and 3 G/s). Cerebral deoxy-hemoglobin increased significantly (max: +30 +/- 6% of control, P < 0.01) only during the 0.1 G/s run. At G-LOC, CBV changes were not related to G-onset rate (P = 0.30; mean change: 32 +/- 6% of control). We conclude that cerebral ischemia is the main mechanism in the occurrence of G-LOC. PMID- 12565143 TI - Effect of 5-hydroxyindole on ethanol potentiation of 5-hydroxytryptamine (5-HT)3 receptor-activated ion current in NCB-20 neuroblastoma cells. AB - We examined the effect of 5-hydroxyindole (5-HI) on the potentiation of 5 hydroytryptamine (5-HT)(3) receptor function by ethanol (EtOH) so as to study whether EtOH potentiates channel function through increasing activation or blocking desensitization. We measured 5-HT(3) receptor current using a whole-cell voltage clamp technique with a method of rapid drug application in NCB-20 neuroblastoma cells. The 5-HI, an agent that block receptor desensitization, increased the decay time constant (tau), not the peak of 5-HT(3) receptor mediated currents induced by 10 microM 5-HT. EtOH did not change the peak amplitude and tau of the current induced by 10 microM 5-HT. Coapplication of EtOH and 5-HI with 5-HT caused no increase in the peak currents, but tau was further increased. Therefore, a further block in desensitization could be induced by 5 HI, despite the presence of EtOH. These results indicate that EtOH potentiates 5 HT(3) receptor function, with these effects due at least in part by increasing channel activation rather than by blocking desensitization. PMID- 12565144 TI - Difference in the representation of cardinal and oblique contours in cat visual cortex. AB - Optical imaging based on intrinsic signals was used to analyze the structure of orientation column in area 18 of cat visual cortex. Recordings were obtained in 12 adult cats during presentation of contours in various orientations. In order to quantitatively evaluate the size of the response area, a method combining both information on the amplitude of the changes and the statistical significance of the changes was proposed. The results showed that the proportion of the cortical area activated reached a peak during the presentation of horizontal or vertical contours, whereas the areas activated by oblique contours were relatively smaller. Cardinal contours (0 degrees and 90 degrees ) activated 50.63 +/- 1.88% (mean +/- SE) of the cortical area in the recording site, whereas oblique contours (45 degrees and 135 degrees ) activated only 45.25 +/- 2.02% of the area. This difference in area was statistically significant (t = 2.38, d.f. = 11, and P < 0.05). The results demonstrated a difference in the representations of the cardinal and oblique contours in area 18 of the cat visual cortex. PMID- 12565145 TI - Mutation screening of the human period 2 gene in bipolar disorder. AB - We tested whether the human period 2 gene (hper2), one of the essential components of the circadian oscillator, might have influence on bipolar disorder. We screened 88 bipolar disorder patients and 127 controls, all of Japanese origin. Screening in the casein kinase I epsilon (CKIepsilon) binding region of hper2, which was previously reported in familial advanced sleep-phase syndrome patients, with polymerase chain reaction amplification revealed four polymorphisms. One of the four polymorphisms had an amino acid substitution of a serine at 662 with a glycine (S662G). The frequencies of the S662G allele and genotypes on patients with bipolar disorder were very low and had no difference from those in controls. Polymorphism on the CKIepsilon binding region of hper2 gene which was previously reported, is unlikely to play an important role in the development of bipolar disorder. PMID- 12565146 TI - Prevalence of TAU mutations in an Italian clinical series of familial frontotemporal patients. AB - Frontotemporal dementia (FTD) is a clinical entity grouping different diagnostic conditions. FTD can occur in a sporadic form; however in 30-50% of cases a familial form of FTD has been observed. Mutations in the TAU gene were associated to familial FTD linked to chromosome 17. Our aim was to investigated the proportion of FTD cases attributable to TAU gene mutations in an Italian clinical series. We analyzed 38 patients with FTD; of these, 13 had a positive family history of FTD. All TAU gene exons and flanking intronic regions were sequenced. In our familial FTD sample the estimation of TAU gene mutations accounted for a relative low prevalence (7.6%); based on our results we could argue the existence of other mutations in regulatory regions in the TAU gene or, on the other hand, other genes might be responsible for the most cases of familial FTD. PMID- 12565147 TI - Orientation of human glioblastoma cells embedded in type I collagen, caused by exposure to a 10 T static magnetic field. AB - We investigated the preferred orientation of human glioblastoma cells (A172) following exposure to static magnetic fields (SMF) at 10 Tesla in the presence or absence of collagen. A172 cells embedded in collagen gel were oriented perpendicular to the direction of the SMF. A172 cells cultured in the absence of collagen did not exhibit any specific orientation pattern after 7 days of exposure to the SMF. Thus we succeeded in evoking the magnetic orientation of human glioblastoma cells by exposure to the SMF. Our results suggest that the orientation of glioblastoma cell processes may be due to the arrangement of microtubules under the influence of magnetically oriented collagen fiber. PMID- 12565148 TI - A convenient approach to the generation of multiple internal control DNA for a panel of real-time PCR assays. AB - Real-time polymerase chain reaction (PCR) assays allow convenient detection and quantitation of virus-derived nucleic acids in clinical specimens. When specimens are assayed for the presence of virus-derived nucleic acids against external standards, sample adequacy is not monitored. This can be achieved by using internal controls that are co-amplified with the virus-specific DNA in competitive PCR. Each of the various real-time PCR assays in a routine clinical laboratory requires its specific internal control. In order to complement a panel of virus-specific real-time PCR assays with internal controls, a convenient approach is described to generate the several internal controls within single DNA fragment. By applying composite primer technology, PCR primer sequences used in real-time PCR assays were added in 5' and 3' of a stretch of heterologous DNA during consecutive preparative PCRs. The heterologous DNA was used for internal control specific detection by e.g. FRET-hybridisation probes. The presented example of such a multiple internal control DNA contained five internal controls for five competitive LightCycler-PCR assays. All five PCR products derived from the multiple internal control DNA were detected with a single pair of specific FRET-hybridisation probes. The example described proved useful in real-time PCR assays specific for the detection of EBV-, CMV-, VZV- HSV-, and HBV-DNA on the LightCycler instrument. This methodology should enable laboratories to conveniently complement their panel of existing real-time PCR assays with a single multiple internal control DNA. PMID- 12565149 TI - Immunofluorescent detection and quantitation of hepatitis A virus in sewage treatment effluent and on agri-food surfaces using scanning confocal microscopy. AB - An immunofluorescent (IF) assay was developed for specific detection and quantitation of hepatitis A virus (HAV) in sewage treatment effluent or attached to stainless steel, copper, high density polyethylene and polyvinyl chloride surfaces. Polyclonal antibodies were produced in rabbits and characterized for specificity to HAV. Purified anti-HAV antibodies were used in combination with ALEXA-anti-rabbit conjugate and confocal microscopy for detection and quantitation of HAV in effluent samples. Using this immunological approach, as little as 2 x 10(5) PFU/ml were detected and the signal generated was proportional to the concentration of HAV up to 2 x 10(8) PFU/ml. Counts obtained by IF assay were highly correlated with those of the SYBR Green II nucleic acid labelling method (r(2)=0.995) and conventional plaque assay (r(2)=0.988). The IF assay described is rapid (3 h), sensitive, specific to HAV and suitable for qualitative as well as quantitative studies. PMID- 12565150 TI - Characterization of a recombinant Newcastle disease virus expressing the green fluorescent protein. AB - A recombinant Newcastle disease virus (NDV) expressing the green fluorescent protein (GFP) was generated by applying reverse genetics techniques. The GFP open reading frame flanked by NDV transcription start and stop sequences was inserted between the fusion (F)- and hemagglutinin-neuraminidase genes in a full-length cDNA clone of NDV. This plasmid transcribing antigenome RNA was cotransfected with helper plasmids expressing viral nucleoprotein, phosphoprotein and large protein into cells stably expressing T7 RNA polymerase. The rescued virus was first propagated in embryonated eggs and the allantoic fluid was used to infect cells. Northern blot analysis of RNA isolated from infected cells demonstrated the proper transcription of the introduced GFP-mRNA. The appearance of GFP in live infected cells confirmed further the recovery of a recombinant NDV (rNDVGFP1) expressing the reporter gene. The expression of the heterologous gene was maintained stably for at least five passages in embryonated eggs. The replication kinetics in embryonated eggs and pathogenicity in chickens of rNDVGFP1 did not differ significantly from that of the parent virus. Using GFP autofluorescence, virus infected cells could be tracked easily in native preparations, organ explants and primary tracheal cell cultures. Taken together, these data demonstrate the use of GFP-expressing recombinant NDV for analysis of NDV dissemination and pathogenesis and indicate the potential usefulness of NDV as a vaccine vector. PMID- 12565151 TI - Preliminary attempts to biolistic inoculation of grapevine fanleaf virus. AB - Biolistics has been studied to inoculate grapevine fanleaf virus (GFLV), a Nepovirus, to its natural woody host, Vitis sp., and its herbaceous host, Chenopodium quinoa. At first, bombardment conditions for in vitro and greenhouse grown plants were set using the uidA reporter gene. The infectious feature of the cartridges was then evaluated by studying infection of C. quinoa plants. Systemic infection was obtained with either GFLV particles or RNA extracts in experimental conditions which gave also the highest transient uidA gene expression. Concerning grapevine, our results indicate that extrapolation to this plant is difficult. In only 1 out of 8 independent bombardment experiments done with GFLV and 41B, we were able to detect the virus in freshly bombarded leaves. Similarly, later after bombardment, Pol mRNAs were detected once, at days 7 and 14 only. Incubating the plants in darkness, as suggested in the literature, or using Rupestris Saint Georges, an indicator for GFLV presence, did not yield any improvement. Finely, our observations suggest that detection of GFLV in bombarded grapevine tissues by immunological or molecular techniques remains a limiting factor, probably due to an excess of inhibitory compounds released during the biolistic process. PMID- 12565152 TI - Differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in formalin-fixed paraffin-embedded tissues by multiplex RT nested PCR and comparison with in situ hybridization. AB - Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) were detected and differentiated in formalin-fixed, paraffin-embedded tissues from experimentally and naturally infected pigs by multiplex reverse transcription-nested polymerase chain reaction (RT-nPCR). The results of this new method were compared with in situ hybridization. A method based on xylene deparaffinization followed by proteinase K digestion yielded RNA of a suitable quality for reliable and consistent multiplex RT-nPCR analyses. PEDV and TGEV cDNAs were detected in jejunal tissues from experimentally and naturally infected pigs by multiplex RT-nPCR. Distinct positive signals for PEDV and TGEV were also detected in the same jejunal tissues by in situ hybridization. The rate of conformity between multiplex RT-nPCR and in situ hybridization was 100% for the detection of PEDV and TGEV in formalin-fixed paraffin-embedded jejunal tissues. PMID- 12565153 TI - Quantitative analysis of LP-BM5 murine leukemia retrovirus RNA using real-time RT PCR. AB - Murine AIDS (MAIDS) develops in susceptible mouse strains after infection with the LP-BM5 murine leukemia virus (MuLV) complex that contains a mixture of defective (BM5def) and replication-competent viruses. While the BM5def virus is the causative agent in MAIDS, the replication-competent viruses in LP-BM5, including ecotropic MuLV (BM5eco), are required for BM5def propagation and thus function as helper viruses. We describe quantitative real-time RT-PCR assays for RNA encoded by the BM5def and BM5eco components of LP-BM5. The assays were used to standardize better the input doses of LP-BM5 viruses across viral preparations and to quantify BM5def and BM5eco gag RNA levels in spleen and blood cells from MAIDS-susceptible and -insusceptible infected mice. Spleens of MAIDS-susceptible infected mice harbored approximately similar levels of BM5def gag RNA as infected spleens of mice that are insusceptible to MAIDS due to lack of CD40. In contrast, the same infected spleens of CD40-deficient mice contained substantially higher (up to 10-fold) levels of BM5eco gag RNA compared with susceptible controls. Similar to that seen in spleen, infected blood of CD40-deficient mice contained similar levels of BM5def gag as susceptible strains, but increased levels (up to threefold) of BM5eco gag RNA. The assays described below can be used to characterize better the contributions of different functional viral components of the LP-BM5 mixture to the development of MAIDS. PMID- 12565154 TI - Cloning and expression of the guinea pig cytomegalovirus glycoprotein B (gB) in a recombinant baculovirus: utility for vaccine studies for the prevention of experimental infection. AB - The guinea pig cytomegalovirus (GPCMV) is unique among the cytomegaloviruses of small mammals, insofar as during pregnancy it crosses the placenta, causing infection of the fetus. Although the guinea pig model is well suited to vaccine studies, the lack of cloned, recombinant forms of immunogenic GPCMV proteins, such as envelope glycoproteins, has hindered experimental evaluations of subunit immunization for prevention of fetal disease. Since the glycoprotein B (gB) is a major target of neutralizing antibody responses, the GPCMV gB was cloned and expressed in a recombinant baculovirus. A recombinant was generated which expressed gB, truncated at codon 692, upstream of the putative transmembrane domain. Processing and expression of the recombinant protein, designated Bac-gB, was assessed, and the protein was characterized immunologically. Anti-gB antibodies were immunoreactive with Bac-gB by enzyme linked immunosorbent assay (ELISA) and immunoblot assay. Immunoprecipitation with polyclonal anti-GPCMV antisera identified protein species of 120, 80 and 30 kDa by reducing SDS-PAGE, suggesting that authentic cleavage and processing of Bac-gB occurred in insect cells. Sera from guinea pigs immunized with lectin-column purified native glycoproteins had high ELISA titers to Bac-gB. Recombinant GPCMV gB expressed in insect cells should prove useful in defining correlates of protective immunity in the GPCMV congenital infection model. PMID- 12565155 TI - A rapid and convenient variant of fusion-PCR to construct chimeric flaviviruses. AB - So far, full-length cDNAs of chimeric flaviviruses have been constructed by restriction-enzyme cleavage of the gene(s) to be exchanged or by fusion-PCR of two amplified PCR fragments. The construction of a chimeric flavivirus by a faster and more convenient variant of the standard fusion-PCR is reported. A Modoc/yellow fever chimeric virus was engineered in which the structural prM and E genes of yellow fever virus 17D were replaced by the homologous genes of Modoc virus. In two PCR steps, a fusion was made between the 3' end of the C gene of yellow fever virus and the 5' end of the prM gene of Modoc virus, and between the 3' end of the E gene of Modoc virus and the 5' end of the NS1 gene of yellow fever virus. For each of the two fusions between yellow fever and Modoc virus, a standard PCR was performed to amplify a short fragment with one overlapping end that could be used as one of the primers in the subsequent (fusion) PCR. PMID- 12565156 TI - Defective influenza A virus generated entirely from plasmids: its RNA is expressed in infected mouse lung and modulates disease. AB - Naturally produced defective influenza virus has antiviral activity and, in sufficient amount, can protect mice from lethal influenza, irrespective of the virus subtype causing the disease. However, such defective virus preparations contain many undefined defective RNA sequences, and it is thus not possible to establish dose-response relationships. To address this situation, we have transfected DNA encoding a cloned defective RNA into Vero cells along with the 17 A/WSN (H1N1) plasmids required for infectious helper virus, and produced molecularly cloned defective virus. Here we used POLI-220 that expresses a 445 nt defective RNA isolated from a mouse-protective defective equine H3N8 virus, and POLI-317 that expresses a 585 nt defective RNA from an avian H7N7 virus. Both originate from genomic segment 1. Virus preparations were UV-irradiated selectively to destroy virus infectivity but not the activity of the defective RNAs, and adult mice were inoculated intranasally with defective virus and WSN (H1N1) challenge virus (10 LD(50)). Defective POLI-220 and POLI-317 RNAs were detected readily in infected lung tissue by RT-PCR, but these Vero cell preparations did not modulate disease. However, after a single passage in embryonated eggs, defective POLI-220 and POLI-317 viruses significantly delayed the onset of disease and death in WSN-infected mice, although did not affect final mortality. Direct PCR sequencing confirmed the identity of mouse-passaged defective RNAs and showed that none had undergone any sequence changes. With this advance it will now be possible to study the interference phenomenon in vivo with defective viruses carrying a defined defective RNA. PMID- 12565157 TI - DNA replicative functions of highly-expressed, codon-optimized human papillomavirus proteins E1 and E2. AB - Human papillomavirus (HPV) DNA replication requires functional HPV early (E) proteins E1 and E2. To determine the biological activity of HPV 16 E1 and E2 mutant proteins under consideration as vaccine candidates, we developed a sensitive real time PCR assay that monitors HPV origin-of-replication-driven DNA synthesis. The assay was used to determine the DNA replicative functions of highly-expressed, codon-optimized HPV 16 E1 and E2 wild type and mutant proteins in transient transfections. Under the assay conditions, the HPV 16 E1 mutant (W439R, G482D) did not support HPV origin-driven DNA synthesis. In contrast, however, an HPV 16 E2 mutant bearing an E39A substitution, reported previously to be severely compromised for DNA replication, was found to be reduced only two fold in activity and, therefore, considered not sufficiently inactivated for use in vaccines that depend on endogenous protein expression. PMID- 12565158 TI - Comparison of electron microscopic techniques for enumeration of endogenous retrovirus in mouse and Chinese hamster cell lines used for production of biologics. AB - Murine myeloma and Chinese hamster ovary cells are used widely in the manufacture of recombinant proteins for biopharmaceuticals. However, rodent cell lines express endogenous retrovirus, which necessitates appropriate design of purification processes to remove virus in excess of the calculated maximum retroviral load. Currently, electron microscopy is the method of choice for determination of retroviral titre in bulk harvest. In this study we compared three electron microscopy techniques to determine retroviral titre in bulk harvest. These were direct negative stain, negative stain after sucrose-density purification and thin section electron microscopy of pelleted supernatant. The study demonstrated that the level of C-type retrovirus associated with cells was predictive of the viral load in cell culture supernatants. The most accurate method for quantifying viral load was direct counting, followed by thin section of pelleted supernatant and negative stain after sucrose concentration. The most practical method was thin section of resuspended pelleted supernatant, which gave improved detection limits. PMID- 12565159 TI - Phenotypic and genotypic methods for the detection of herpes simplex virus serotypes. AB - Typing of herpes simplex virus (HSV) into its serotypes plays a major role in epidemiology and management of reactivation. To develop and evaluate a polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) was employed using Hae III and Taq I against neutralization test, allele-specific PCR and DNA sequencing for the detection of HSV serotypes. Neutralization test, allele-specific PCR, DNA sequencing and PCR-based RFLP were applied simultaneously to 2 standard strains (HSV-1 and HSV-2) and 23 clinical isolates. PCR-based RFLP was applied further to 20 culture negative PCR positive clinical specimens. The 179 bp product of the clinical isolates and specimens amplified using the type-common primers of HSV was subjected to DNA sequencing and PCR based RFLP. Allele-specific PCR was absolutely specific and highly sensitive. All the typing methods differentiated concordantly 23 clinical isolates into 12 HSV-1 and 11 HSV-2. DNA sequencing did not reveal any nucleotide variations within the serotypes among the isolates sequenced. PCR-based RFLP typed a further 20 culture negative clinical specimens into 15 HSV-1 and 5 HSV-2. PCR-based RFLP was a reliable, less laborious and cost-effective molecular biological tool for the determination of HSV serotypes both for the clinical isolates and culture negative specimens. PMID- 12565160 TI - Quantitative-competitive PCR monitoring of viral load following experimental guinea pig cytomegalovirus infection. AB - Human cytomegalovirus (HCMV) is the most common cause of congenital viral infection in the developed world, and can lead to significant morbidity. Animal models of HCMV infection are required for study of pathogenesis, because of the strict species-specificity of cytomegalovirus (CMV). Among the small animal CMV models, the guinea pig CMV (GPCMV) has unique advantages, in particular its propensity to cross the placenta, causing disease in utero. In order to develop quantitative endpoints for vaccine and antiviral therapeutic studies in the GPCMV model, a quantitative-competitive PCR (qcPCR) assay was developed, based on the GPCMV homolog of the HCMV UL83 gene, GP83. Optimal amplification of GPCMV DNA was observed using primers spanning a 248 base pair (bp) region of this gene. A 91 bp deletion of this cloned fragment was generated for use as an internal standard (IS) for PCR amplification. Standard curves based upon the fluorescent intensity of full-length external target to IS were compared with signal intensity of DNA extracted from blood and organs of experimentally infected guinea pigs in order to quantify viral load. Viral load in newborn guinea pigs infected transplacentally was determined and compared with that of pups infected with GPCMV as neonates. Viral loads were highest in pups infected as neonates. The most consistent isolation and highest quantities of viral DNA were observed in liver and spleen, although viral genome could be readily identified in brain, lung, and salivary gland. Viral load determination should be useful for monitoring outcomes following vaccine studies, as well as responses to experimental antiviral agents. PMID- 12565161 TI - Detection of immunoglobulin G to Crimean-Congo hemorrhagic fever virus in sheep sera by recombinant nucleoprotein-based enzyme-linked immunosorbent and immunofluorescence assays. AB - Crimean-Congo hemorrhagic fever virus is a tick-borne virus that causes severe hemorrhagic symptoms with an up to 50% mortality rate in humans. Wild and domestic animals, such as sheep, cattle and goats, are the reservoirs. The recombinant nucleoprotein-based Crimean-Congo hemorrhagic fever virus antibody detection systems for sheep sera were developed by enzyme-linked immunosorbent assay (ELISA) and an indirect immunofluorescence assay techniques. The samples used for evaluation were 80 sera collected from sheep in a Crimean-Congo hemorrhagic fever-endemic area (western part of the Xinjiang Uygur Autonomous Region) and 39 sera collected from sheep in a disease-free region (Shandong province, eastern China). The ELISA and indirect immunofluorescence assay using recombinant nucleoprotein of the virus proved to have high sensitivity and specificity for detecting the immunoglobulin G antibodies to the virus in sheep sera. Within this limited number of samples, the recombinant nucleoprotein-based ELISA and indirect immunofluorescence assay are considered to be useful tools for seroepidemiological study of virus infections in sheep sera. PMID- 12565162 TI - Diagnostic rapid tests for acute hantavirus infections: specific tests for Hantaan, Dobrava and Puumala viruses versus a hantavirus combination test. AB - Hantaviruses infecting humans in Eurasia include Hantaan, Seoul, Puumala and the closely related Dobrava and Saaremaa viruses. These viruses are causative agents of hemorrhagic fever with renal syndrome (HFRS), which is recognized as a severe health care problem in several countries. Diagnostics of hantavirus infections relies on serology, performed principally with enzyme immunoassay (EIA) or immunofluorescence assay (IFA). We developed four 5-min immunochromatographic IgM antibody tests for diagnostics of acute Puumala, Dobrava and Hantaan virus infections and a similar combination test to detect all Eurasian pathogenic hantavirus infections. We evaluated the assays using 100 fingertip blood samples collected randomly from Finnish volunteers, 28 confirmed hantavirus IgM-negative sera, and 77 sera from patients with acute infections of various hantaviruses. The specificities and sensitivities of the Puumala-, Dobrava- and Hantaan virus specific tests varied from 96 to 100%, whereas, the combination test showed 96% specificity and 80 to 93% sensitivity. Cross-reactions were observed commonly between the Dobrava and the Hantaan virus tests, but only rarely between the Puumala and the Hantaan virus, or the Puumala and the Dobrava virus, tests. Altogether, the rapid tests showed less cross-reactivity than the respective EIA tests. According to the results, the performance of these tests meets well the requirements for diagnostic use. Nevertheless, the specific one-antigen tests were markedly more sensitive than the combination test. However, if optimized, a combination test would be suitable for regions where several hantaviruses circulate. PMID- 12565163 TI - Evaluation of the NucliSens Basic Kit assay for detection of Norwalk virus RNA in stool specimens. AB - Norwalk-like viruses (NLVs) are a genetically diverse group of human caliciviruses that are the most common cause of epidemic gastroenteritis and are detected typically in stool by reverse transcription (RT)-PCR or electron microscopy (EM). The application of a rapid nucleic acid sequence-based amplification (NASBA) assay for the detection of NLV RNA in stool is described using the NucliSens Basic Kit. Primers and probes for the NLV Basic Kit assay were based on the RNA polymerase region of the prototype NLV, Norwalk virus (NV) genome and could consistently detect 10(4) RT-PCR detectable units of NV RNA in a stool filtrate. When compared directly with RT-PCR on a dilution series of NV stool filtrate, the NucliSens Basic Kit assay was equally sensitive. Cross reactivity studies with a representative panel of other enteric pathogens were negative. When applied to 15 stool specimens from NV-challenged volunteers, the NASBA Basic Kit application for NV detection yielded 100% sensitivity, 50% specificity, and 67% concordance, using RT-PCR as the 'gold standard'. Despite the specificity of the NASBA primer/probe sequences for NV, other representatives from both NLV genogroups I and II could be detected by the Basic Kit assay in outbreak stool specimens, although the results were inconsistent. Our results suggest that the NucliSens Basic Kit assay provides a rapid and sensitive alternative to RT-PCR for detecting NV RNA in stool specimens. However, improvements in test specificity and primer design will be needed before the assay can be used routinely in the clinical setting. PMID- 12565164 TI - Colony establishment and maintenance of the eriophyid wheat curl mite Aceria tosichella for controlled transmission studies on a new virus-like pathogen. AB - High plains disease (HPD) is of serious economic concern for wheat and corn production, but little is known about the virus-like causal agent. In the field, HPD is often associated with Wheat streak mosaic virus (WSMV) and both pathogens are transmitted by the same eriophyid wheat curl mite, Aceria tosichella Keifer. The objective of this study was to develop methods for establishing and maintaining HPD-transmitting wheat curl mite colonies for their use in studies on HPD. Towards this goal, mite colonies from a mixed infection source were separated into colonies either (i). not viruliferous; (ii). only transmitting WSMV; or (iii). only transmitting HPD. Maintenance of these colonies required strictly separated incubator facilities and adaptation of mite-suitable transfer techniques to permit frequent passages of mites to healthy plants. The established colonies provided reliable sources of infective material to study the progression of HPD and/or WSMV in plants using sensitive immuno-detection assays. In conclusion, we have developed reliable methods with a poorly studied arthropod vector to examine the biology and properties of a new virus-like disease. PMID- 12565165 TI - Single-tube fluorescent product-enhanced reverse transcriptase assay with Ampliwax (STF-PERT) for retrovirus quantitation. AB - A TaqMan fluorescent probe-based product enhanced reverse transcriptase (RT) assay is described in which the RT and polymerase chain reaction (PCR) steps are set-up in a single tube, in two compartments separated by Ampliwax (designated as single-tube fluorescent product-enhanced reverse transcriptase assay (STF-PERT)). This simplification of the two-step method resulted in increased assay reproducibility and handling efficiency while maintaining the sensitivity of the PERT assay (<10 virions). The STF-PERT assay can be used to quantitate low amounts of retrovirus in clinical and research materials and to evaluate retrovirus contamination in cell substrates and biological products in human use. PMID- 12565166 TI - Epigenetic gene silencing in cancer initiation and progression. AB - Hypermethylation of CpG islands, an epigenetic event that is not accompanied by changes in DNA sequence, represents an alternative mechanism to deletions or mutations to inactivate tumor suppressor genes. Recent evidence supports the notion that CpG island hypermethylation, by silencing key cancer-related genes, plays a major causal role in cancer. However, a long-standing issue in the field is the sequence of molecular events leading to epigenetic gene silencing. A new model has been proposed that chromatin remodeling, as a result of histone deacetylation and methylation, is the primary event in abrogating transcriptional initiation; subsequently, CpG island hypermethylation establishes a permanent state of gene silencing. Accumulating evidence indicates that CpG island hypermethylation is an early event in cancer development and, in some cases, may precede the neoplastic process. Because of their heritable nature, hypermethylated CpG islands leave 'molecular footprints' in evolving cancer cells and can be used as molecular markers to reconstruct epigenetic progression during tumorigenesis. Furthermore, hypermethylated CpG islands are proving to be useful for molecular classification of different cancer types. PMID- 12565167 TI - Sera from cancer patients contain two oscillating ECTO-NOX activities with different period lengths. AB - ECTO-NOX protein's are cell surface-associated and growth-related hydroquinone oxidases with both protein disulfide-thiol interchange activity and the capacity to oxidize NAD(P)H. The activities of these ECTO-NOX proteins are not steady state but fluctuate to create a repeating pattern of oscillations. Two forms of ECTO-NOX activities have been distinguished. The constitutive ECTO-NOX (CNOX), is hormone responsive and refractory to quinone-site inhibitors. A tumor-associated NOX (tNOX) is unregulated, refractory to hormones and growth factors and responds to quinone-site inhibitors. CNOX proteins are widely distributed and exhibit oscillations in enzymatic activity with a period length of 24 min. tNOX proteins are cancer specific and exhibit oscillations with a period length of about 22 min. Our findings now demonstrate the presence of the novel oscillating tNOX activity in sera of patients with cancer whereas the constitutive NOX of non cancer cells is present in sera of both cancer patients and healthy volunteers. We conclude that ECTO-NOX proteins in sera exhibit oscillatory characteristics similar to those of ECTO-NOX forms of the cell surface. PMID- 12565168 TI - Circadian rhythm of tumor promotion in the two-stage model of mouse tumorigenesis. AB - The question of whether cancer risk is influenced by time-of-day exposure to potentially carcinogenic agents was approached in this study by exposing mouse skin to a single initiating dose of 7,12-dimethylbenz [A-]anthracene, followed by a 12 week regime of bi-weekly skin treatments with the tumor promoter, 12-0 tetradecanoyl-phorbol acetate (TPA), given at four different circadian clock times (CCTs). Tumor incidence, average number of tumors per mouse and tumor size showed a dominant circadian component with an acrophase occurring at 23:00 h CCT. Pre-treatment with all trans-retinoic acid, prior to bi-weekly TPA promotion, reduced tumor incidence, average number and size of tumors per animal by greater than 80%, but did not suppress the underlying circadian rhythm of sensitivity to TPA-induced tumor formation. PMID- 12565169 TI - Genotoxicity of advanced glycation end products in mammalian cells. AB - In patients with chronic renal failure, cancer incidence is enhanced. Since levels of advanced glycation end products (AGEs) are markedly elevated in renal insufficiency, we investigated potential effects of various AGEs on structural DNA integrity in tubule cells. The comet-assay was employed, a method based on the computer-aided microscopic analysis of single cells after electrophoretic separation of their nuclear DNA. Incubation of pig kidney LLC-PK1-cells for 24 h with AGE-BSA (AGE-bovine serum albumin), carboxymethyllysine-BSA as well as methylglyoxal-BSA resulted in a significant increase in DNA damage. Pretreatment of the cells with the proteases trypsin and bromelain abolished the AGE-induced comet-formation. This is in agreement with the idea that the observed genotoxicity of AGEs could be receptor-mediated and that proteases inactivate the extracellular domain of the receptor for AGEs. Binding of AGEs to the RAGE receptor leads to an increased intracellular formation of active oxygen species, which are known to induce DNA damage. It is concluded that AGEs induce genotoxicity in tubule cells, which may be involved in the enhanced cancer development in advanced kidney diseases. PMID- 12565170 TI - Resveratrol is a potent inducer of apoptosis in human melanoma cells. AB - Resveratrol is a plant polyphenol found in grapes and red wine. It has been found to have beneficial effects on the cardiovascular system. Resveratrol also inhibits the growth of various tumor cell lines in vitro and inhibits carcinogenesis in vivo. In this study we examined the effect of resveratrol on growth of two human melanoma cell lines. We found that this plant polyphenol inhibited growth and induced apoptosis in both cell lines, with the amelanotic cell line A375 being more sensitive. The potential involvement of different MAP kinases in the action of resveratrol was also examined. Although resveratrol did not alter the phosphorylation of p38 or JNK MAP kinases in either cell line, it induced phosphorylation of ERK1/2 in A375, but not in SK-mel28 cells. These results suggest that in vivo studies of the effect of resveratrol on melanoma are warranted and that this plant polyphenol might have effectiveness as either a therapeutic or chemopreventive agent against melanoma. PMID- 12565171 TI - Pentacyclic triterpenes from Chrysobalanaceae species: cytotoxicity on multidrug resistant and sensitive leukemia cell lines. AB - Plants are known as important source in the search for new anti-cancer agents. Cytotoxicity-guided fractionation of leaves and fruits from Licania tomentosa Bench and leaves from Chrysobalanus icaco L. resulted in the isolation of betulinic, oleanolic and pomolic acids. These triterpenoids inhibited the growth and induced apoptosis of K562, an erythroleukemia cell line. Most importantly, they also inhibited the proliferation of Lucena 1, a vincristine-resistant derivative of K562 that displays several multidrug resistance (MDR) characteristics. Taken together, our findings emphasize the anti-tumor activity of these triterpenes on leukemia cell lines and call attention to their potential as anti MDR agents. PMID- 12565172 TI - Calagualine inhibits nuclear transcription factors-kappaB activated by various inflammatory and tumor promoting agents. AB - Calagualine derived from the fern of the genus Polypodium, commonly called calaguala, has had clinically documented medicinal uses in South America and Spain and been shown to block tumor metastasis, proliferation, and inflammation, all known to require the activation of nuclear transcription factor-kappaB (NF kappaB). Therefore, we investigated the effect of calagualine on NF-kappaB activation induced by various inflammatory and tumor promoting agents. Calagualine blocked tumor necrosis factor (TNF)-induced activation of NF-kappaB through inhibition of phosphorylation and degradation of IkappaBalpha, an inhibitor of NF-kappaB. The effects of calagualine were not cell type-specific, as it blocked TNF-induced NF-kappaB activation in a variety of cells. NF-kappaB dependent reporter gene transcription activated by TNF was also suppressed by calagualine. The TNF-induced NF-kappaB activation cascade involving TNFR1-TNF receptor-associated death domain-TNF receptor-associated factor 2 (TRAF2)-NF kappaB-inducing kinase (NIK)-IkappaBalpha kinase was interrupted at the TRAF2 and NIK sites by calagualine, which would account for its suppression of NF-kappaB reporter gene expression. Calagualine blocked NF-kappaB activation induced by phorbol ester and lipopolysaccharide. Overall our results indicate that calagualine inhibits activation of NF-kappaB and this may provide a molecular basis for calagualine's ability to suppress inflammation and tumorigenesis. PMID- 12565173 TI - Polymorphisms of XRCC1 and XRCC3 genes and susceptibility to breast cancer. AB - Mammalian cells are constantly exposed to a wide variety of genotoxic agents from both endogenous and exogenous sources. Genetic variability in DNA repair may contribute to human cancer risk. We used a case-control study design (162 cases and 302 controls) to test the association between three amino acid substitution variants of DNA repair genes (XRCC1 Arg194Trp, XRCC1 Arg399Gln, and XRCC3 Thr241Met) and breast cancer susceptibility. We found a weak association between the XRCC1 194Trp allele and breast cancer risk (adjusted odds ratio (OR)=1.98; 95% confidence interval (CI)=0.85-4.63). We also found a potential gene-gene interaction between the XRCC1 194Trp allele and XRCC3 241Met allele and breast cancer risk (adjusted OR=8.74; 95% CI=1.13-67.53). Although larger studies are needed to validate the study results, our data suggest that amino acid substitution variants of XRCC1 and XRCC3 genes may contribute to breast cancer susceptibility. PMID- 12565174 TI - Nuclear expression of YB-1 protein correlates with P-glycoprotein expression in human breast carcinoma. AB - Drug resistance is a common clinical problem in cancer treatment. The overexpression of P-glycoprotein appears to be closely associated with multi-drug resistance to anticancer agents. YB-1 binds to the Y-box in the promoter region of MDR1, which encodes P-glycoprotein. We evaluated the correlation between nuclear YB-1 and P-glycoprotein expression and other molecules, such as hormonal receptors, angiogenic factors, immune factors, and methalloprotease in 63 human breast cancers. Nuclear YB-1 expression had a significant correlation with P glycoprotein and PgR expression, and also with CD68 grade, concerning accumulation of tumor associated macrophages. This series did not demonstrate P glycoprotein and nuclear YB-1 expression might be one of the useful prognostic marker of breast cancer. PMID- 12565175 TI - Differentially expressed genes in gastric tumors identified by cDNA array. AB - Using cDNA fragments from the FAPESP/lICR Cancer Genome Project, we constructed a cDNA array having 4512 elements and determined gene expression in six normal and six tumor gastric tissues. Using t-statistics, we identified 80 cDNAs whose expression in normal and tumor samples differed more than 3.5 sample standard deviations. Using Self-Organizing Map, the expression profile of these cDNAs allowed perfect separation of malignant and non-malignant samples. Using the supervised learning procedure Support Vector Machine, we identified trios of cDNAs that could be used to classify samples as normal or tumor, based on single array analysis. Finally, we identified genes with altered linear correlation when their expression in normal and tumor samples were compared. Further investigation concerning the function of these genes could contribute to the understanding of gastric carcinogenesis and may prove useful in molecular diagnostics. PMID- 12565176 TI - p16 Hypermethylation in the early stage of hepatitis B virus-associated hepatocarcinogenesis. AB - Abnormality of the p16 expression is involved in the pathogenesis of hepatocellular carcinoma (HCC), and hypermethylation of p16 gene is known as a major p16 inactivation mechanism. Cirrhotic nodule (CN) is now regarded as a preneoplastic lesion that is frequently associated with microscopic foci of HCC through dysplastic nodules (DNs). This observation clearly supports a multistep hepatocarcinogenesis from CNs through DNs. We thus examined the methylation status of p16 gene in HCCs surrounded by DNs and CNs to define the significance of p16 hypermethylation in the early stage of hepatocarcinogenesis. We tested 24 hepatitis B virus (HBV)-associated CNs, 37 DNs, and 18 HCCs within DNs that were microdissected from paraffin-embedded tissue sections. Frequency of p16 hypermethylation was significantly high in HCCs within DNs (15/18. 83.3%) and it increased from CNs (15/24. 62.5%) through DNs (26/37, 70.3%). Interestingly, 11 out of 12 (91.7%) HCC associated with methylation-positive DNs revealed hypermethylation of p16, and 18 out of 23 (78.2%) DNs associated with methylation positive CNs showed p16 hypermethylation. These data suggest that p16 hypermethylation in the early stages, CNs and DNs may predispose to HCC. In addition, p16 methylation status of five cell lines with or without HBV infection was examined to test whether the high frequency of hypermethylation is related to HBV infection. HBV-infected cell lines were exclusively methylation-positive. These data suggest that high frequency of hypermethylation may be associated with hepatitis B virus infection. PMID- 12565177 TI - The minimum LOH region defined on chromosome 17p13.3 in human hepatocellular carcinoma with gene content analysis. AB - Hepatocellular carcinoma (HCC) is one of the most common human cancers in Asia. Previous studies have shown that in addition to aberrations of the p53 gene on chromosome 17p13.1, other gene(s) on chromosome 17p13.3 may also play a role in HCC. To detect the status of loss of heterozygosity (LOH) in HCC and to determine the minimum region of LOH on 17p13.3, we analyzed 22 paired HCC and non-cancerous liver samples with 14 polymorphic markers plus TP53 (p53 gene) as a comparison. The data revealed a high level of LOH (>68%) in a minimum region between D17S1866 and D17S1574, spanning over a 1.5 Mb region. Genomic library screening using markers in the region has resulted in the isolation of a cluster of BAC/PAC clones. We created a physical map in this region. Using large-scale genome sequencing, gene annotation, cDNA screening, and exon trapping, we identified 17 known genes and 13 novel genes in the minimum region. The function of these genes was analyzed and the possibility of several putative tumor suppressor genes was discussed. PMID- 12565178 TI - Expression of p53R2, newly p53 target in oral normal epithelium, epithelial dysplasia and squamous cell carcinoma. AB - Recently, the p53R2 gene has been isolated and shown to play a crucial role in DNA repair after DNA damage. The p53R2 gene encodes the p53 inducible ribonucleotide reductase small subunit 2 homologue, which is part of the p53 pathway. However, the function of p53R2 in human cancer is still unclear. We investigated p53R2 mRNA expression in human oral normal epithelium, epithelial dysplasias and squamous cell carcinomas (SCCs). Surgical or biopsy-proven specimens of 10 normal epithelium, 48 epithelial dysplasias and 63 SCCs were collected in our department. Then, p53R2 was identified by in situ hybridization to visualize and localize the expression of specific mRNAs. The authors examined the p53 gene mutation by polymerase chain reaction-single strand conformation polymorphism analysis. p53, mdm2, p21(WAF1/CIP1) and Ki-67 expression was detected by immunohistochemistry. p53R2 expression was detected in none of ten normal epithelium (0%), ten of 48 dysplasias (20.8%) and 33 of 63 SCCs (52.4%). In oral SCC, the expression of p53R2 was significantly associated with tumor size, lymph node metastasis and histological differentiation (P=0.014, 0.046 and 0.022, respectively). p53R2 expression was significantly associated with p53 abnormality in epithelial dysplasia and SCC (P=0.034 and 0.009, respectively). Of 63 patients, 37 received preoperative radiochemotherapy. p53R2 mRNA expression was significantly associated with the pathologic response to radiochemotherapy (P=0.031). This study suggested that p53R2 expression could be associated with oral carcinogenesis. The presence of p53R2 mRNA expression would be a predictive factor for tumor development, tumor cell differentiation and the sensitivity to radiochemotherapy in oral SCC. PMID- 12565179 TI - Intravesical diamorphine for bladder spasm. PMID- 12565180 TI - A case of cannabinoid rotation in a young woman with chronic cystitis. PMID- 12565181 TI - Intermittent nasogastric drainage under sedation for unresponsive vomiting in terminal bowel obstruction. PMID- 12565182 TI - Angina pain precipitated by a continuous subcutaneous infusion of ketamine. PMID- 12565183 TI - Re: Mirtazapine in cancer patients. PMID- 12565184 TI - Description and predictors of direct and indirect costs of pain reported by cancer patients. AB - The purpose of this study was to describe direct and indirect costs associated with pain in cancer patients and to examine potential predictors of these costs. The study surveyed cancer outpatients about direct costs resulting from pain related hospitalizations, emergency department visits, physician office visits, and use of analgesic medications and indirect costs related to money spent on pain-related transportation, complementary methods to improve pain management, educational materials, over-the counter medication, domestic support, and childcare. Furthermore, the study examined age, marital status, race, income level, pain severity, pain interference, and presence of breakthrough pain as predictors of direct and indirect costs. Three hundred and seventy-three cancer outpatients were sampled. One hundred and forty-four cancer patients (39%) reported experiencing cancer-related pain and completed the study questionnaires. Seventy-six percent (76%) of the patients had experienced at least one pain related cost, resulting in an average monthly direct cost of US$ 891/month per patient. Sixty-nine percent (69%) of patients had experienced some type of direct medical cost due to pain, resulting in an average total direct pain-related cost of US$ 825/month per patient. Fifty-seven percent (57%) of patients reported incurring at least one indirect pain-related expense for an average indirect cost of US$ 61/month per patient. Higher pain intensity, greater pain interference, and presence of breakthrough pain predicted higher direct and indirect medical expenses. Younger age and lower income level also predicted higher direct medical expenses. PMID- 12565185 TI - Physicians' interactions with health care teams and systems in the care of dying patients: perspectives of dying patients, family members, and health care professionals. AB - This study investigated the specific physician skills required to interact with health care systems in order to provide high quality care at the end of life. We used focus groups of patients with terminal diseases, family members, nurses and social workers from hospice or acute care settings, and physicians. We performed content analysis based on grounded theory. Groups were interviewed. Two domains were found related to physician interactions with health care systems: 1) access and continuity, and 2) team communication and coordination. Components of these domains most frequently mentioned included taking as much time as needed with the patient, accessibility, and respect shown in working with health team members. This study highlights the need for both physicians and health care systems to improve accessibility for patients and families and increase coordination of efforts between health care team members when working with dying patients and their families. PMID- 12565186 TI - Effect of cancer pain on performance status, mood states, and level of hope among Taiwanese cancer patients. AB - The purposes of this study were: 1) to compare performance status, mood states, and level of hope between patients with cancer pain and patients without cancer pain; and 2) to determine the relationships of pain intensity and pain interference with daily life to performance status, mood states, and level of hope. A total of 233 Taiwanese cancer patients with pain and 251 without pain participated. The self report instruments consisted of the Chinese version of the Profile of Mood States (POMS) short form, the Chinese version of the Herth Hope Index, the Brief Pain Inventory-Chinese version (BPI-C), the Chinese version of the Karnorfsy Performance Scale (KPS), and a demographic questionnaire. The major findings of this study were that cancer patients with pain reported significantly lower levels of performance status and higher levels of total mood disturbance than did cancer patients who did not experience pain after controlling for sex, disease stage, and recruitment site. In addition, patients with cancer pain experienced significantly more anger, fatigue, depression, confusion, and lethargy than did patients without pain after controlling for sex, disease stage, and recruitment site. Among patients with pain, pain intensity was significantly correlated with performance status and mood state, but not with level of hope. Pain interference with daily life was significantly correlated both with performance status, mood state, and level of hope. Pain intensity and pain interference were significantly correlated with each mood state as well as with total mood disturbance. This study has demonstrated the effect of cancer pain on patients' physical, psychological, and spiritual life and has supported the multidimensional notion of the cancer pain experience in Taiwanese patients. PMID- 12565187 TI - Driving ability under long-term treatment with transdermal fentanyl. AB - Clinical experience shows that neuropsychological side effects due to opioid therapy usually decrease during the first weeks of therapy. However, the effect of long-term treatment with transdermal fentanyl on complex activities, such as driving, is not yet clear. In a prospective trial, patients with continuous noncancer pain, who had received stable doses of transdermal fentanyl for at least 2 weeks, completed a series of computerized tests to measure attention, reaction, visual orientation, motor coordination and vigilance. Data from 90 healthy volunteers were matched to 30 patients; 9 patients were excluded from the per-protocol analysis because they took additional drugs in violation of the protocol. None of the performance measures for the 21 remaining fentanyl patients was significantly inferior to the controls. We conclude that stable doses of transdermal fentanyl for the treatment of chronic non-cancer pain are not associated with significant impairments in psychomotor and cognitive performance. The threshold for fitness to drive as defined by German law did not differ significantly between the groups. PMID- 12565188 TI - Pain assessment in elderly patients with severe dementia. AB - The purpose of this study was to assess the reliability and validity of facial expressions as pain indicators in patients with severe dementia. Based on interviews with patients who could report pain, we defined characteristics of decubitus ulcers associated with reports of pain during dressing changes. We then evaluated 9 patients who had ulcers with these characteristics but were unable to communicate verbally because of severe dementia. We videotaped their facial expressions before and during their decubitus ulcer dressing change. We showed the videotape segments, in random order, to 8 medical students and 10 nurses. The 18 viewers were asked to infer the presence or absence of pain based on their observations of the patients' facial expressions and vocalizations. The dressing change of decubitus ulcers extending beyond the subcutaneous tissue, covering an area of at least 9 cm(2), and with a moist surface, was always reported as painful by study patients able to report (95% confidence interval of 69-100%). The intraclass correlation coefficient for the answers of the 18 viewers evaluating each videotape segment for the presence of pain was 0.64. Sensitivity, specificity, and positive and negative predictive values of viewers' ratings of facial expressions and vocalizations as a measure of the presence of pain were: 0.70, 0.83, 0.90, and 0.81. The intraclass correlation coefficient for the answers rating pain intensity was only 0.10, indicating only slight agreement beyond chance. Assuming dressing changes of ulcers reported as painful by communicative patients are also painful in non-verbal severely demented patients, clinician observations of facial expressions and vocalizations are accurate means for assessing the presence of pain, but not its intensity, in patients unable to communicate verbally because of advanced dementia. PMID- 12565189 TI - Does mammography hurt? AB - The documented incidence of pain associated with screening mammography varies from 1% to 62%. Some researchers suggest that pain may undermine compliance with screening mammography. As a part of a quality improvement project, we have surveyed women undergoing mammography in 2 centers in Jerusalem to identify the prevalence, severity, and duration of mammography-associated pain, demographic risk factors, and the degree that this may undermine compliance with breast cancer screening. A 23-item questionnaire was administered to 399 women (32% at the Shaare Zedek Medical Center [SZMC] and 68% at the Rachel Nash Comprehensive Breast Clinic [HALA]). Of the total, 77% of the women reported that the procedure was painful. Of those reporting pain, 60% described pain intensity as moderate or severe. In 67%, the pain resolved within 10 minutes. By univariate analysis, the only significant predictor for pain during mammography was cyclic breast pain (P = 0.053). No significant correlation was identified for age, breast size, pre mammography counseling, and examination center (SZMC vs. HALA). The prevalence of pre-mammography counseling or explanation was low (51%). Despite that, 61% of the respondents expected that mammography would be painful. Indeed, most of those who anticipated pain reported that the actual severity was not greater than the anticipated severity. Even among women who reported pain of moderate or greater severity, less than 5% expressed preference to receive pre-emptive analgesia prior to their next mammogram. A substantial minority of women acknowledged that the experience of their mammography invoked reactions that may impend future compliance; 26% reported anxiety and 12% reported pain as factors that may interfere with ongoing compliance with regular mammographic screening. These data serve to emphasize the need for appropriate pre-test counseling and suggest a possible role for post-test debriefing to address those factors which may interfere with future test compliance. PMID- 12565190 TI - Stability and reliability of detection thresholds for human A-Beta and A-delta sensory afferents determined by cutaneous electrical stimulation. AB - Activity in primary afferent fibers that usually mediate fine touch can evoke sensations of pain in conditions in which there is sensitization of central neurons. Input from these large diameter Abeta afferents may also sustain and exacerbate these central mechanisms. The role of these fibers in clinical pain syndromes can be evaluated by applications of electrical stimuli that preferentially activate Abeta axons. This study assessed the stability and reliability of a method of electrical stimulation (ES) useful for clinical evaluation. Monopolar constant-current rectangular pulses were delivered to 5 equi-spaced sites on the volar aspect of the left forearm along a transverse line 5 cm distal to the antecubital crease. Current intensity was gradually increased to determine detection threshold and pain detection threshold. This study determined: 1) Effect of pulse duration (1, 2, and 5 msec); 2) the variation of detection threshold and pain threshold over repeated stimulation; 3) the effect of electrode position with respect to distance from the trunk of underlying ulnar or median nerves; and 4) the effect of re-positioning the electrode on variability of detection threshold and pain threshold. There was no significant variability over time for either detection threshold (DT) or pain threshold (PT) at any of the 3 pulse durations tested. There was also no significant effect on variability of shifting the electrode between sites, nor was there a significant difference in variability between sites when placed either over or adjacent to peripheral nerves. Under simulated clinical conditions of electrode re positioning, the mean detection threshold in 300 trials and ten subjects was 0.30 mA with an overall standard error of 0.007, standard errors of 0.014 over the 10 subjects, 0.003 over the 6 trials, and 0.012 over the 5 locations. Similarly, mean pain threshold in these 300 trials was 3.24 +/- 0.093, with standard errors of 0.12 over the 10 subjects, 0.023 over the 6 trials, and 0.13 over the 5 locations. Mean ratio of pain threshold divided by detection threshold ratio was 10.9 +/- 0.25 with a range of 2.0-28.3. Single pulse, constant current electrical stimulation of the skin at threshold levels is a quantifiable and reliable sensory method that is repeatable within and between testing sessions. Our results suggest that in skin unaffected by allodynia, a ratio of the two sensory thresholds (pain threshold and detection threshold) of less than 2.0 is uncommon. We propose that, in the presence of mechanical allodynia, a pain threshold/detection threshold of less than 2.0 suggests that altered central nervous system processing of Abeta input may contribute to allodynia. PMID- 12565191 TI - Relationships among morphine metabolism, pain and side effects during long-term treatment: an update. AB - The two metabolites of morphine, morphine-3-glucuronide (M3G) and morphine-6 glucuronide (M6G), have been studied intensively in animals and humans during the past 30 years in order to elucidate their precise action and possible contribution to the desired effects and side effects seen after morphine administration. M3G and M6G are formed by morphine glucuronidation, mainly in the liver, and are excreted by the kidneys. The metabolites are found in the cerebrospinal fluid after single as well as multiple doses of morphine. M6G binds to opioid receptors, and animal studies have demonstrated that M6G may be a more potent analgesic than morphine. Results from human studies regarding the analgesic effect of M6G are not unanimous. The potency ratio between systemic M6G and morphine in humans has not been settled, but is probably lower than previously assumed. Hitherto, only a few studies have found evidence for a contributory effect of M6G to the overall effects observed after morphine administration. Several studies have demonstrated that administration of M6G is accompanied by fewer and a milder degree of opioid-like side effects than observed after morphine administration, but most of the studies have used lower doses of M6G than of morphine. M3G displays very low affinity for opioid receptors and has no analgesic activity. Animal studies have shown that M3G may antagonize the analgesic effect of morphine and M6G, but no human studies have demonstrated this. M3G has also been connected to certain neurotoxic symptoms, such as hyperalgesia, allodynia and myoclonus, which have been observed after administration of M3G or high doses of morphine in animals. The symptoms have been reported sporadically in humans treated primarily with high doses of morphine, but the role of M3G in eliciting the symptoms is not fully elucidated. PMID- 12565195 TI - Zearalenone induces male germ cell apoptosis in rats. AB - Zearalenone (ZEA), a nonsteroidal estrogenic mycotoxin, is known to cause toxicity of testis in male rats. To investigate whether apoptosis is involved in ZEA-induced testicular toxicity and to identify the stage and target germ cell type, 10-week-old Sprague-Dawley male rats were treated with a single intraperitoneal (i.p.) dose of ZEA (5 mg/kg) and euthanized at 3, 6, 12, 24, or 48 h subsequently. Histopathologically, germ cell degeneration was found at stages I-VI 12 h after dosing. Degenerating germ cells were shown to undergo apoptosis as revealed by in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The frequency of TUNEL-labeled germ cells increased in a stage-specific manner, the peak frequency gradually progressing at stages I-VI of seminiferous tubules with time after dosing, suggesting that the damaged germ cells, especially spermatogonia and spermatocytes, gradually underwent the processes leading to apoptosis. DNA laddering on gel electrophoresis was apparent 12 h after dosing. The results demonstrated that a single dose of ZEA induces testicular germ cell apoptosis in a time-dependent and stage-specific pattern. This study has established that apoptosis is the principal mechanism contributing to germ cell depletion and testicular atrophy following ZEA exposure. PMID- 12565197 TI - Inhibition of cyclophosphamide-induced teratogenesis by beta-ionone. AB - Beta-ionone (BI) is a degraded (C 13) sesquiterpene found in plant essential oils. It has been used in the synthesis of perfume chemicals and vitamin A. Recently, it was reported that BI is a rather potent in vitro inhibitor of CYP2B1 catalysed reactions in rat liver microsomes. The present study was performed to investigate whether inhibition of CYP2B1 reactions by BI could lead to an attenuation of cyclophosphamide (CP)-induced embryotoxicity in the rat. In a preliminary experiment, a dose-dependent prolongation of pentobarbital sleeping time in male and female Wistar rats suggested that BI inhibits CYP2B1 in vivo as well. In a second experiment, rats were treated by gavage with BI (0, 250, 500, 750 or 1000 mg/kg body wt) 45 min prior to a subcutaneous injection of either CP (7.5 mg/kg body wt) or its vehicle (saline) on day 11 of pregnancy. BI alone, at the highest dose tested, caused a high proportion of resorptions. Lower doses of BI, however, clearly attenuated CP-induced embryolethality and teratogenicity. These results seem to support the view that, as far as rats are concerned, CYP2B1 plays an important role in the conversion of CP into its embryolethal and teratogenic metabolites. PMID- 12565196 TI - Fatty acid release and oxidation are factors in lipoxygenase inhibitor-induced apoptosis. AB - MK886, an inhibitor of 5-lipoxygenase activating protein (FLAP), and the lipoxygenase (LOX) inhibitors baicalein and nordihydroguaiaretic acid (NDGA), induce apoptosis by mechanisms independent of both LOX and FLAP. One possible mechanism for these agents is through an effect on the binding of fatty acids to LOX and fatty acid binding proteins resulting in increased intracellular levels of unbound fatty acids, particularly arachidonic acid (AA), that in turn, activate apoptosis signaling pathways either directly or following oxidation. In FL5.12 murine pro-B lymphocytic cells, exogenous fatty acids induced apoptosis proportional to their degree of unsaturation. MK886, baicalein, and NDGA significantly enhanced the release of [3H]-AA two to threefold within 2 h and induced apoptosis by 8 h. Neither MK886-induced AA release, nor apoptosis were affected by quinacrine, a phospholipase A2 inhibitor. The presence of peroxides 1 h after treatment of FL5.12 cells with these agents was evident by a two to threefold increase in the ferrous oxidation-xylenol orange (FOX) assay as well as dichlorofluorescein fluorescence measured with flow cytometry. Isoprostane formation, an additional index of lipid peroxidation, was increased threefold by 2 h, and fourfold at 4 h after MK886 or baicalein, but not after NDGA. Antioxidants were able to protect against NDGA-induced apoptosis but had no effect on baicalein and resulted in enhanced apoptosis with MK886. These data support the hypothesis that release of fatty acids and generation of oxidized species contribute to apoptosis induced by these LOX inhibitors, but that more complex mechanisms are likely involved. PMID- 12565198 TI - Changes in peroxisome proliferator-activated receptor gamma-regulated gene expression and inhibin/activin-follistatin system gene expression in rat testis after an administration of di-n-butyl phthalate. AB - Changes in gene expression regulated by peroxisome proliferator-activated receptor gamma (PPARgamma) and in gene expression related to the inhibin/activin follistatin system in the rat testis induced by a single oral administration of di-n-butyl phthalate (DBP) (8.6 mmol/kg) were examined and compared with those in the control rats using reverse-transcriptase polymerase chain reaction (RT-PCR). The increase in cytochrome P450 4A1 mRNA, which is regulated by PPARalpha, was significant, but not so profound as the increase of P450 4A1 mRNA in the liver. In contrast, a remarkable increase in the mRNA level of plasminogen activator inhibitor-1 (PAI-1) was found in the testis, suggesting the activation of PPARgamma. The substantial increase in PAI-1 may be related to the disruption of spermatogenesis. On the other hand, significant suppression of the mRNA level of inhibin beta(B) and elevation in the mRNA level of follistatin, an activin binding protein, were observed after the DBP-administration. Activin B, a homodimer of inhibin beta(B), is known to stimulate spermatogonial proliferation. The present results suggest that the suppression of spermatogenesis resulting from the changes in the expression of genes involved in the inhibin/activin follistatin system is one of the mechanisms of the testicular atrophy induced by DBP. PMID- 12565199 TI - Effects of pretreatment with verapamil on the toxicokinetics of soman in rabbits and distribution in mouse brain and diaphragm. AB - The effects of verapamil on the elimination of soman in rabbit blood and distribution in mouse brain and diaphragm by determining the concentration of P( )soman using the chirasil capillary gas chromatographic analysis method were studied in order to study the effects of verapamil on the metabolic detoxification of soman. Verapamil (10 mg kg(-1), im, 30 min before soman administration) could significantly reduce the concentration of P(-)soman in rabbit blood at 15, 60, 90, 120, 180 and 240 s after soman injection (43.2 microg kg(-1), iv) as compared to soman-treated control animal respectively. Toxicokinetics parameters showed verapamil could increase clearance rate from 20.8+/-1.51 to 44.3+/-7.0 ml kg(-1)s(-1) and reduce AUC of P(-)soman from 2.08+/ 0.151 to 0.996+/-0.172 mg s l(-1). For experiments in mice, verapamil could reduce the concentration P(-)soman in diaphragm from 74.7, 70.5, 88.7 to 41.1, 39.0, 49.3 ng g(-1) at the time of 30, 90, 120 s after intoxication of soman subcutaneously vs. soman control respectively, but it had no influence on the concentration of free P(-)soman in brain. Verapamil accelerated the elimination of P(-)soman in the rabbits blood and reduced the distribution of P(-)soman in the mouse diaphragm. PMID- 12565201 TI - Astroglial CYP1B1 up-regulation in inflammatory/oxidative toxic conditions: IL 1beta effect and protection by N-acetylcysteine. AB - The present work aims to determine the relevance of an astrocytoma cell line U373 MG, for assessing the role of some astroglial cytochrome P450 in neurotoxicity and neuroprotection. CYP1B1, CYP2C8, CYP2C9, CYP2D6, CYP2J2, CYP2E1 and CYP4A11 mRNA were detected by reverse transcriptase-polymerase chain reaction in control U373 MG cell cultures. Among them we focused on CYP1B1 expression. After 48 h treatment with a range of concentrations of interleukin-1beta (1, 5, 10 ng/ml) used to simulate stress conditions, CYP1B1 mRNA expression was enhanced in a dose dependent way. This increased expression was followed 24 h later by an increase in protein level, determined by Western-blot. N-acetylcysteine (NAC) partially inhibited this effect both on the mRNA and protein levels. As CYP1B1 activates procarcinogenic compounds to reactive metabolites, an increase in this P450 isoform will participate to toxic consequences of an inflammatory/oxidative stress. NAC will prevent this deleterious effect. PMID- 12565200 TI - Effect of cisplatin and cobalt chloride on antioxidant enzymes in the livers of Lewis lung carcinoma-bearing mice: protective role of heme oxygenase. AB - Changes in the activities of antioxidant enzymes superoxide dismutase, catalase (CAT), glutathione peroxidase and heme oxygenase (HO) and changes in lipid peroxidation and reduced glutathione (GSH) levels were measured in the livers of control and Lewis lung carcinoma (LLC)-bearing mice 24 h after a single injection of cisplatin or CoCl(2). Treatment with cisplatin induced the same degree of lipid peroxidation and GSH depletion as did CoCl(2) but the antioxidant enzymes were differently involved in cisplatin- and cobalt-induced oxidative stress responses. In cobalt-treated mice the activities of these enzymes were either inhibited or not changed significantly and only the HO activity was increased (5 fold) as a main protective enzyme. In cisplatin-treated animals the antioxidant enzymes were activated but the enhancement of HO and CAT was greater in LLC inoculated mice. It is suggested that these two enzymes represent the protective response against cisplatin toxicity in the livers of tumor-bearing animals. PMID- 12565202 TI - Assessment of reproductive and fertility effects of amitraz pesticide in male mice. AB - Forty adult male Swiss mice were exposed to tap water containing 0, 40, 80, or 160 ppm amitraz for 12 weeks. Based on fluid consumption the mice received an average of 0, 5.42+/-0.47, 10.56+/-0.97, and 20.39+/-2.17 mg/kg/day amitraz, respectively. The average body weights gains and fluid consumption were significantly decreased in males exposed to amitraz pesticide. Fertility was significantly reduced in male mice ingesting 10.56+/-0.97 or 20.39+/-2.17 mg/kg/day amitraz in that the number of females impregnated by them was significantly reduced. The number of viable fetuses was significantly reduced in females mated with males that ingested 10.56+/-0.97 or 20.39+/-2.17 mg/kg/day amitraz. A significant increase in the total number of resorptions and the number of females with resorptions was observed in females impregnated with the exposed males. Absolute testis weight was significantly decreased at 10.56+/-0.97 mg/kg concentration. The weight of the epididymis was decreased in test males ingested 20.39+/-2.17 mg/kg amitraz. The seminal vesicles weights were significantly increased in male mice ingested 10.56+/-0.97 or 20.39+/-2.17 mg/kg/day amitraz. Similarly, the preputial gland weights were increased in males that ingested 5.42+/-0.47 or 10.56+/-0.97 mg/kg and decreased in males ingested 20.39+/-2.17 mg/kg amitraz. Testicular sperm counts and daily sperm production were significantly decreased in males that ingested 10.56+/-0.97 or 20.39+/-2.17 mg/kg/day amitraz. Epididymal sperm counts were significantly decreased in exposed male's at 10.56+/-0.97 or 20.39+/-2.17 mg/kg amitraz. These results strongly suggest that exposure to amitraz pesticide have an adverse effect on the fertility and reproductive system of male mice. PMID- 12565203 TI - Magnetic resonance imaging of short T2 components in tissue. AB - The most widely used clinical magnetic resonance imaging techniques for the diagnosis of parenchymal disease employ heavily T(2)-weighted sequences to detect an increase or decrease in the signal from long T(2) components in tissue. Tissues also contain short T(2) components that are not detected or only poorly detected with conventional sequences. These components are the majority species in tendons, ligaments, menisci, periosteum, cortical bone and other related tissues, and the minority in many other tissues that have predominantly long T(2) components.The development and clinical application of techniques to detect short T(2) components are just beginning. Such techniques include magic angle imaging, as well as short echo time (TE), and ultrashort TE (Ute) pulse sequences. Magic angle imaging increases the T(2) of highly ordered, collagen-rich tissues such as tendons and ligaments so signal can be detected from them with conventional pulse sequences. Ute sequences detect short T(2) components before they have decayed, both in tissues with a majority of short T(2) components and those with a minority. In the latter case steps usually need to be taken to suppress the signal from the majority of long T(2) components. Fat suppression of different types may also be helpful. Once signal from short T(2) components has been detected, different pulse sequences can be used to determine increases or decreases in T(1) and T(2) and study contrast enhancement. Using these approaches, signals have been detected from normal tissues with a majority of short T(2) components such as tendons, ligaments, menisci, periosteum, cortical bone, dentine and enamel (the latter four tissues for the first time) as well as from the other tissues in which short T(2) components are a minority. Some diseases such as chronic fibrosis, gliosis, haemorrhage and calcification may increase the signal from short T(2) components while others such as loss of tissue, loss of order in tissue and an increase in water content may decrease them. Changes of these types have been demonstrated in tendonopathy, intervertebral disc disease, ligament injury, haemachromatosis, pituitary perivascular fibrosis, gliomas, multiple sclerosis and angiomas. Use of these techniques has reduced the limit of clinical detectability of short T(2) components by about two orders of magnitude from about 10 ms to about 100 micros. As a consequence it is now possible to study tissues that have a majority of short T(2) components with both "bright" and "dark" approaches, with the bright (high signal) approach offering options for developing tissue contrast of different types, as well as the potential for tissue characterization. In addition, tissues with a minority of short T(2) components may demonstrate changes in disease that are not apparent with conventional heavily T(2)-weighted sequences. PMID- 12565204 TI - MRI in the differential diagnosis of a sellar mass. AB - Whilst pituitary adenomas are the commonest cause of a sellar mass, there are a number of other neoplastic, infectious, inflammatory, developmental and vascular aetiologies that should be considered by the radiologist. We discuss and illustrate these lesions and indicate the various magnetic resonance imaging features that are helpful in formulating an appropriate differential diagnosis. PMID- 12565205 TI - Neuroimaging for non-accidental head injury in childhood: a proposed protocol. AB - Non-accidental head injury (NAHI) is a major cause of neurological disability and death during infancy. Radiological imaging plays a crucial role in evaluating craniospinal injury, both for guiding medical management and the forensic aspects of abusive trauma. The damage sustained is varied, complex and may be accompanied by an evolving pattern of brain injury secondary to a cascade of metabolic and physiological derangements. Regrettably, many cases are poorly or incompletely evaluated leading to diagnostic errors and difficulties in executing subsequent child care or criminal proceedings. It is evident, from cases referred to the authors, that imaging protocols for NAHI are lacking (or only loosely adhered to, if present) in many centres throughout the U.K. Future research in this field will also be hampered if there is a lack of consistent and reliable radiological data. There is no nationally agreed protocol for imaging NAHI. We propose such a protocol, based upon a wide experience in the medical management of child abuse and extensive involvement in the medicolegal aspects of NAHI. PMID- 12565206 TI - Is the presence of mammographic comedo calcification really a prognostic factor for small screen-detected invasive breast cancers? AB - AIM: It has been suggested that the use of traditional prognostic factors such as histological grade and lymph node stage are not reliable predictors of outcome for small (<15 mm diameter) screen-detected breast cancers. It has also been suggested that the presence of mammographic comedo (casting) microcalcifications may be an important prognostic factor. We have determined prognostic factors for small screen-detected cancers in a univariate and multivariate fashion. MATERIALS AND METHODS: Histological grade, lymph node stage, invasive tumour size and vascular invasion status was obtained on 161 consecutive patients with primary invasive breast cancer <15 mm diameter, detected by screening. The Nottingham Prognostic Index was calculated for each patient. The mammograms were assessed for the presence of microcalcifications, these being classified as either comedo or non-comedo. Survival data was also obtained. RESULTS: Overall survival in this group of patients was excellent with only 12 patients (7.5%) dying from their breast cancer (mean follow-up 7.5 years). Univariate analysis showed a significant association between lymph stage and outcome (chi(2)=9.68, P=0.008). No significant association was demonstrated between the presence of comedo calcification and survival. Multivariate analysis confirmed lymph node stage as the only independent prognostic factor for these small screen-detected breast cancers (chi(2)=7.18, P=0.007). There were significant associations between the presence of comedo calcification on the screening mammogram and high histological grade and small tumour size. CONCLUSION: Although the overall outcome for small screen-detected breast cancers (<15 mm diameter) is excellent, the presence of lymph node metastases is associated with a significant reduction in long-term survival. The presence of mammographic comedo calcification is not an independent prognostic factor, but is closely related to histological grade. PMID- 12565207 TI - Can radiographers read screening mammograms? AB - AIM: To evaluate the ability of radiographers to read screening mammograms in the National Health Service Breast Screening Programme (NHSBSP). MATERIALS AND METHODS: Two radiographers read a test set of 1000 screening mammograms previously reported by a consultant radiologist. Three radiographers then acted as a second reader for 54,000 screening mammograms, their recall results are presented. Four consultant radiologists and three film reading radiographers were timed while reading 2500 mammograms each. RESULTS: When reading the test mammograms the two radiographers recalled all the cancers previously detected by the radiologist at the original screen read. They also recalled 32/90 women who subsequently presented with interval cancers. As a second reader the radiographers had similar recall and cancer detection rates to the radiologists (P>0.05). Double reading detected 9% more cancers. The radiographers take the same length of time to film read as radiologists (P>0.05). CONCLUSION: Radiographers are able to read screening mammograms at least as well as radiologists and do not take longer to do so. PMID- 12565208 TI - Bowel wall thickening in patients with Crohn's disease: CT patterns and correlation with inflammatory activity. AB - AIM: To assess CT patterns of bowel wall thickening in patients with Crohn's disease and to correlate these patterns with inflammatory activity. MATERIALS AND METHODS: We conducted a retrospective review of 58 helical abdominal CT scans of 53 patients with pathologically proven Crohn's disease. CT patterns of thickened bowel wall were divided into four types based on patterns of mural stratification and enhancement: type A, multilayered mural stratification; type B, two layers with strong mucosal enhancement and prominent low-density submucosa; type C, two layers without strong mucosal enhancement; and type D, homogeneous enhancement. We evaluated CT findings of the bowel and adjacent structures. We also reviewed pathologic features and clinical data to determine inflammatory activity. RESULTS: Fifty-five (95%) of 58 CT examinations showed bowel wall thickening. Of these 55 CT scans, type A pattern was found in 33 (60%), type B in 10 (18%), type C in five (9%), and type D in seven (13%). CT scans with type A showed significantly more wall thickening than those with either type C or type D. Histology revealed 43 cases with active disease and 12 with quiescent appearance. Thirty of 33 CT scans with type A and all 10 with type B were classified as acute disease, and three of five with type C and six of seven with type D as quiescent. CONCLUSION: In patients with Crohn's disease, CT patterns of bowel wall thickening correlated with inflammatory activity. Thickened bowel wall with layering enhancement is predictive of acute disease, and that of homogeneous enhancement suggests quiescence. PMID- 12565209 TI - Ultrasound-guided endocavitary drainage of pelvic abscesses: technique, results and complications. AB - AIM: To evaluate the experience in our institution with ultrasound-guided transrectal and transvaginal (endocavitary) drainage of pelvic abscesses. MATERIALS AND METHODS: Eighteen patients (four male, 14 female; mean age 55 years, range 30-78 years) presenting with pelvic abscesses were referred to our institution for therapeutic drainage over a 4 year period. Patients received broad-spectrum antibiotics prior to drainage, which was performed by either the transvaginal or transrectal route under ultrasound guidance. Patients were given sedo-analgesia in the form of midazolam and fentanyl and local anaesthesia was also employed. Eight French catheters were inserted into the abscess cavities, and patients were subsequently monitored on a daily basis by a member of the interventional radiology team until such time as it was deemed appropriate to remove the catheter. RESULTS: Eighteen catheters were placed in 17 patients, and transvaginal aspiration alone was performed in one patient. Drainage was successful in 16 of 17 patients, but a transgluteal approach was ultimately required in the remaining patient to enable passage of a larger catheter into an infected haematoma. The mean duration of drainage was 5 days, mean time to defervesce 2 days. Spontaneous catheter dislodgement occurred in four patients associated with straining, but this did not have any adverse effect in three of the four patients. CONCLUSION: Endocavitary drainage is an effective method of treatment for pelvic abscesses. Spontaneous catheter dislodgement does not affect patient outcome. PMID- 12565210 TI - Invasive ductal carcinoma arising in a breast hamartoma: two case reports and a review of the literature. PMID- 12565211 TI - Sewing needles as foreign bodies in the liver and pancreas. PMID- 12565212 TI - A case of multiple linseeds mimicking polyposis coli on double contrast barium enema. PMID- 12565213 TI - Imaging in prostate cancer. PMID- 12565683 TI - Volume estimation of prefrontal cortical subfields using MRI and stereology. AB - The objective of this protocol was to provide a rapid, neurofunctionally relevant alternative to region-drawing or automated gyral/sulcal-based techniques. The Cavalieri method and point counting [e.g. Br. J. Radiol. 73 (2000) 679] were used in conjunction with a previously established parcellation methodology [Arch. Gen. Psychiatry 57 (2000) 761] to estimate the volumes of anatomically defined subfields of the prefrontal cortex (PFC) based on landmarks visible on T(1) weighted magnetic resonance (MR) images. Ten participants (n=5 healthy adults; n=5 patients) were studied. Regional PFC volume estimates derived from point counting methods were reproducible between raters (Intraclass Correlations (ICC)=0.92-0.95) and repeatable within rater (ICC=0.93-0.99). Predicted coefficients of error for individual volume estimates were less than 5%. This protocol provides an efficient means of calculating unbiased volume estimates of the PFC with predictable precision for use in both cognitive and clinical studies. PMID- 12565684 TI - A detailed procedure and dissection guide for the isolation of spiral ganglion cells of the guinea pig for electrophysiological experiments. AB - In the present study step-by-step instructions are provided for a preparative technique employed for the removal of the spiral ganglion from the inner ear of the guinea pig. Removal of the temporal bone is followed by opening of the bulla and excision of the modiolus. All major steps of the technique are illustrated with photographs. A procedure to obtain surviving, acutely separated spiral ganglion neurones is also described. By this procedure small tissue pieces are removed from the modiolus which contain the spiral ganglion neurones. The tissue fragments then undergo a mild enzyme treatment (collagenase and pronase). After the enzyme exposure, the tissue pieces are gently triturated, and the isolated cells are allowed to settle. Poly-D-lysine ensured the firm attachment of the spiral ganglion cells to the cover-slips. The application of this adhesive coating seemed to be desirable in functional studies when microelectrode techniques and/or rapid exchange of the extracellular solution were employed. PMID- 12565685 TI - Immunohistochemistry as a tool for topographical semi-quantification of neurotransmitters in the brain. AB - Immunohistochemistry is a powerful tool to detect neurotransmitter (NT) presence in different brain structures with a high spatial resolution. However, it is only scarcely used in quantitative approach due to lack of reproducibility and sensitivity. We developed a protocol of NT detection based on immunohistochemistry and image analysis to show that this approach could also be useful to evaluate NT content variations. We focused our study on the GABAergic system in the cerebellum and measured different accurate parameters, namely the optical density (O.D.), the stained area and the number of immunoreactive cells in each cerebellar cell layer. In order to modify the GABA content, we used gamma vinyl-GABA (GVG), an inhibitor of GABA-transaminase, known to dramatically increase GABA concentration in the central nervous system (CNS) and especially in the cerebellum. We observed a significant increase in the three parameters measured in the molecular and the granular layers of the cerebellum after treatment with GVG, reflecting the well-established increase in GABA content after such a treatment. Therefore, our technical approach allows not only a precise determination of the effects in particular cell layers but also a semi quantification of GABA content variations. This technique could be suitable for monitoring NT variations following any treatment. PMID- 12565686 TI - Methods to optimize the generation of cDNA from postmortem human brain tissue. AB - The analysis of gene transcript levels in postmortem human brain is a valuable tool for the study of neurological and psychiatric diseases. Optimization of the methods of RNA extraction and cDNA generation is particularly important in this application because postmortem human brain tissue is in limited supply and generally yields less RNA than many other human tissues. We compared column extraction and solvent extraction for total RNA, reverse transcription (RT) with random hexamers versus oligo-dT priming, and incubation of the RNA with or without DNase for effect on the cDNA product derived from the same homogenized pool of postmortem human frontal cortex. The total RNA obtained from the solvent method was found to be less stable at room temperature and to contain a higher proportion of non-messenger RNA than that obtained from the column method. Evaluating the RT-PCR results per wet weight of tissue extracted, we found that the signal strength was increased >20-fold by a protocol of Qiagen RNeasy column extraction, random hexamer RT priming and omitting DNase treatment of the RNA. PMID- 12565687 TI - Simultaneous determination of purine nucleotides, their metabolites and beta nicotinamide adenine dinucleotide in cerebellar granule cells by ion-pair high performance liquid chromatography. AB - The method described here allows the quantitative simultaneous determination of adenosine 5'-triphosphate, adenosine 5'-diphosphate, adenosine 5'-monophosphate, adenosine, guanosine 5'-triphosphate, guanosine 5'-diphosphate, guanosine, inosine 5'-monophosphate, inosine, uric acid, xanthine, hypoxanthine and beta nicotinamide adenine dinucleotide by ion-pair high performance liquid chromatography. The chromatographic analysis requires 26 min per sample and allows the separation of the mentioned metabolites in a time as short as 16 min. Primary cultures of rat cerebellar granule cells were incubated in serum-free medium containing 25 mM KCl for 1.5-48 h and their acid extracts were injected onto column. Uric acid, inosine 5'-monophosphate, inosine, beta-nicotinamide adenine dinucleotide, adenosine, adenosine 5'-monophosphate, guanosine 5' diphosphate, adenosine 5'-diphosphate, guanosine 5'-triphosphate and adenosine 5' triphosphate were identified and quantified, while hypoxanthine, xanthine and guanosine were below the detection limit. This method makes use of a single-step sample pre-treatment procedure which allows a greater than 91% recovery of the compounds of interest and provides the assay of the metabolites of interest in little amounts of cell extracts. Therefore, this method is suitable to evaluate the energetic state in a variety of cell types, both under normal and dismetabolic conditions, such as after the induction of apoptosis or necrosis. PMID- 12565688 TI - A functional MRI protocol for localizing language comprehension in the human brain. AB - We describe a functional magnetic resonance imaging (fMRI) protocol to separate activation of areas in the brain associated with language comprehension from sensory areas activated as a result of the presentation of the language stimulus, by comparing cortical activation patterns during the presentation of similar or the same language stimulus via two different sensory modalities (auditory and visual), and identifying the regions of activation that are common to both modalities. The protocol can be implemented on any MR scanner capable of functional imaging, and has proven valuable for the reliable identification of the lateralization and location of language centres in patients being considered for neurosurgical procedures. As well, the method has potential for the study of cortical processing of auditory speech and written language in healthy subjects and in subjects suffering from language disorders. PMID- 12565689 TI - A comparison of (13)C NMR measurements of the rates of glutamine synthesis and the tricarboxylic acid cycle during oral and intravenous administration of [1 (13)C]glucose. AB - 13C-labeled glucose is increasingly used in conjunction with magnetic resonance spectroscopy to measure rates of metabolic pathways in the brain in vivo. Most studies of human subjects have used intravenous infusions to administer the labeled compounds, but the procedure is cumbersome and can be uncomfortable for patients with neurological or psychiatric disorders. It may be possible to improve the practicality of the method by administering the glucose orally instead of intravenously. This report describes the performance and comparison of the oral and intravenous protocols in the same subjects. The conclusion is that oral administration does yield the same result as intravenous administration but with lower precision. That sensitivity of the oral protocol may be improved by several ways that are available today. PMID- 12565690 TI - Temperature control of respiring rat brain slices during high field NMR spectroscopy. AB - Advancing methodologies in high field multinuclear (1)H/(31)P NMR spectroscopy continue to improve the chemical shift precision and signal amplitude sensitivity attainable in ex vivo NMR studies of respiring tissues. Thus it was straightforward in a 14.1-Tesla (600-MHz) system to use the well-known temperature dependence of the chemical shift of water protons to implement a protocol addition in our brain slice studies (350 microm thick, 15 slices per 8 mm NMR tube) of hypoxia. The protocol addition provides for accurate, continuous measurements of brain tissue temperature, a very important variable in studies of oxygen deprivation and metabolism. The basic protocol for loading and maintaining superfused respiring neonatal cerebrocortical slices, similar to protocols described previously by us and others, permitted rapid detection of glycolytic and TCA cycle activity. All brain slices have an 'injury layer' approximately 50 microm thick from mechanical shear. In our protocol the injury layer is confined to one side of the slice, and appears to contribute minimally to NMR spectra. PMID- 12565691 TI - Lead-induced developmental perturbations in hippocampal Sp1 DNA-binding are prevented by zinc supplementation: in vivo evidence for Pb and Zn competition. AB - Zinc finger protein (ZFP) transcription factors are essential for regulation of gene expression in the developing brain. We previously reported that Pb exposure perturbed the DNA-binding of ZFP such as Sp1 and Egr-1 in the cerebellum, which play critical role in CNS development. In this study, we focused on hippocampal Sp1 DNA-binding and mRNA expression in neonatal Pb-exposed animals. The expression pattern of an Sp1 target (NMDAR1) gene was also monitored. To study in vivo and in vitro competition between Pb and Zn, we supplemented animals with Zn, and examined the effects of both metals on hippocampal Sp1 DNA-binding and the DNA-binding of a recombinant Sp1 protein (rhSp1). Tissue metal analysis revealed that only the disposition of Pb in the brain but not its distribution in the blood was influenced by the presence of Zn. The developmental profile of Sp1 DNA binding exhibited a peak on PND 15 which subsequently declined to adult levels. Consistent with earlier studies, Pb exposure produced premature peaks of Sp1 DNA binding on PND 5 which later returned to adult levels. The basal and Pb-induced developmental patterns of Sp1 mRNA departed from its DNA-binding profiles. However, the expression patterns of the NMDAR1 gene were relative to Sp1 DNA binding. Supplementation with zinc provided a protective effect on Pb-induced changes in Sp1 DNA-binding. Moreover, Pb and Zn directly interfered with the DNA binding of rhSp1 in vitro. These data suggest that Pb and Zn can compete both in vivo and in vitro at the zinc finger domain of Sp1 with a consequential effect on Sp1 DNA-binding, subsequent gene expression and brain development. PMID- 12565692 TI - Corn feeding during development induces changes in the number of serotonergic neurons in the raphe nuclei. AB - Serotonin (5-HT) plays a trophic role during brain development; chronic changes in cerebral concentration of this neurotransmitter during the critical stage of development can produce severe damage in the formation of the neural circuits. For the present work a hypoproteic (HYP) diet based on corn (CORN) meal which is deficient in tryptophan (TRY) was given to rats before and during pregnancy, which continued to the offspring until they reached 60 days of age. An isocaloric but hypoproteic diet containing normal amount of TRY, and normal chow (Ch) Purina were given with the same scheme to two groups of rats considered as controls. 5 HT immunohistochemistry was revealed by avidin-biotin complex (ABC) method to quantify serotonergic nerve cells in the nine raphe nuclei. The number of cells immunoreactive to 5-HT immunoreactive (5-HTir) were quantified by means of stereological analysis. Results demonstrated a significant variation in 5-HT expression in the raphe nuclei. Thus, a significant reduction in the number of 5 HTir cells in the rostral raphe nuclei was seen at all ages studied in the animals fed the corn diet, compared to data obtained from the control groups. This decrease was more evident between the postnatal ages of 30 and 60 days. It is concluded that the variations in the available TRY affect the brain cells producing 5-HT and the innervation of their target areas. PMID- 12565693 TI - Neurotrophin-like immunoreactivity in the human pre-term newborn, infant, and adult cerebellum. AB - The immunohistochemical occurrence of the neurotrophin (NT) proteins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-4 (NT-4), and neurotrophin-3 (NT-3) is shown in the pre-term newborn, infant, and adult human post-mortem cerebellum. The NT-like immunoreactive structures were unevenly distributed and showed regional differences among cerebellar lobules and folia. NGF-, NT-4-, and NT-3-positive neuronal perikarya were observed in all specimens examined. At variance with the other neurotrophins, the BDNF antiserum labelled neuronal cell bodies only in newborn life and infancy, as well as extensive nerve fibre systems, whose density increased with age. The NT-antibodies, tested by Western blot on human cerebellum homogenates, revealed immunoreactive bands corresponding to proteins of heterogenous molecular weight. The results obtained provide a first demonstration of the tissue localization of the NTs in the human cerebellum from perinatal to adult age, thus suggesting their involvement in the development, differentiation and maintenance of the cerebellar connectivity. Codistribution of the four NTs or sets of them was observed in cortical and deep nuclei neurons. Multiple trophic roles for NTs, encompassing the classic target derived and local mechanisms of support, are envisaged as significant in development, differentiation, and maintenance of the human cerebellar connectivity. PMID- 12565694 TI - GABA(Abeta2-3) immunoreactive cells in the developing chick retina. AB - Gama-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system (CNS). It has been shown that GABA is an important factor for CNS maturation and that its functions are mainly mediated by GABA(A) receptors. Thus, in order to fully comprehend the role of GABA during development, it is essential to establish the developmental features of the catalytic subunits (beta) of GABA(A) receptor. Here, we determine the ontogenesis and neurogenesis of cells expressing beta2-3 subunits of GABA(A) receptor (GABA(Abeta2-3)) in the chick retina. In the ontogenetic experiments, only the immunohistochemistry for GABA(Abeta2-3) approach was employed. For neurogenesis a double-labeling method (autoradiography and immunohistochemistry) was applied. [H(3)]-thymidine was injected into eggs (2-11 days) and the embryos were sacrificed at embryonic day 19 (E19). GABA(Abeta2-3) immunohistochemistry was processed and then autoradiography was performed. We used a cumulative counting method to quantify the autoradiographic grains. The ontogenesis study revealed that at E9, GABA(Abeta2-3) immunoreactivity was restricted to the inner plexiform layer and the first cell bodies immunoreactive to GABA(Abeta2-3) were seen at E14. Thereafter, the number of cell bodies and the intensity of GABA(Abeta2-3) immunoreactivity increased until the adult pattern was established. The neurogenesis study showed that cells that will express GABA(Abeta2-3) were generated between E6 and E9. In addition, from E7 to E9 the rate of neurogenesis of GABA(Abeta2-3) immunoreactive cells quickly increases. Therefore, the detection of GABA(Abeta2-3) occurred only after the end of generation period of this cell population. PMID- 12565695 TI - Characterization of proliferation and differentiation of EGF-responsive striatal and septal precursor cells. AB - The epigenetic manipulation of precursors may provide data to elucidate the potential interactions among these cells in different brain regions. However, the response to epigenetic signals is modulated by the environment in which the cells are manipulated. Therefore, data regarding the action of a particular factor must be considered in the light of a specific system. To compare septal and striatal precursors, we have tested the effect of nerve growth factor (NGF) on the proliferation and neuronal differentiation of epidermal growth factor (EGF) responsive cells from these brain regions. Precursors were cultivated as 'neuropheres' in serum free medium (SFM) to which NGF was added. NGF did not support the proliferation of EGF-generated precursors so that no differences in the cell magnitude with respect to control cultures were observed. Differentiation of precursors in SFM plus 1% fetal bovine serum (FBS) on poly-D lysine showed that the neuron number was increased two-fold in septal cultures treated with NGF but not in those from striatum. A quantitative evaluation of the soma surface and the number of primary neurites showed differences between both populations of precursor-generated neurons. In addition, we also observed no influence of NGF on these parameters of cellular morphology. Thus, taken together these results seem to indicate that at this developmental stage in which these populations of precursors were isolated, heterogeneities exist between them, which is probably related to their origin and/or functional roles in vivo. PMID- 12565697 TI - Membrane targeting. PMID- 12565696 TI - AMPA receptor potentiation by acetylcholinesterase is age-dependently upregulated at synaptogenesis sites of the rat brain. AB - We have used radioligand binding to synaptic membranes from distinct rat brain regions and quantitative autoradiography to investigate the postnatal evolution of acetylcholinesterase (AChE)-evoked up-regulation of alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) receptors in CNS areas undergoing synaptogenesis. Incubation of synaptosomal membranes or brain sections with purified AChE caused a developmentally modulated enhancement in the binding of [3H]-(S)-AMPA and the specific AMPA receptor ligand [3H]-(S)-5-fluorowillardiine, but did not modify binding to kainate neither N-methyl-D-aspartate receptors. In all postnatal ages investigated (4, 7, 14, 20, 27, 40 days-old and adult rats), AChE effect on binding was concentration-dependent and blocked by propidium, BW 284c51, diisopropylfluorophosphonate and eserine, therefore requiring indemnity of both peripheral and active sites of the enzyme. AChE-mediated enhancement of [3H]-fluorowillardiine binding was measurable in all major CNS areas, but displayed remarkable anatomical selectivity and developmental regulation. Autoradiograph densitometry exhibited distinct temporal profiles and peaks of treated/control binding ratios for different cortices, cortical layers, and nuclei. Within the parietal, occipital and temporal neocortices, hippocampal CA1 field and cerebellum, AChE-potentiated binding ratios peaked in chronological correspondence with synaptogenesis periods of the respective AMPA-receptor containing targets. This modulation of AMPA receptors by AChE is a molecular mechanism able to transduce localized neural activity into durable modifications of synaptic molecular structure and function. It might also contribute to AChE mediated neurotoxicity, as postulated in Alzheimer's disease and other CNS disorders. PMID- 12565698 TI - Renal aquaporin water channels: from molecules to human disease. AB - Following the discovery of the aquaporin-1 water channel in 1991, molecular techniques have been developed to examine the roles of renal aquaporins-1, -2, 3, and -4 in disorders of water balance. This article reviews current knowledge regarding aquaporin function and dysfunction in water-losing and water-retaining states. PMID- 12565699 TI - Physiological and pathophysiological roles of ATP-sensitive K+ channels. AB - ATP-sensitive potassium (K(ATP)) channels are present in many tissues, including pancreatic islet cells, heart, skeletal muscle, vascular smooth muscle, and brain, in which they couple the cell metabolic state to its membrane potential, playing a crucial role in various cellular functions. The K(ATP) channel is a hetero-octamer comprising two subunits: the pore-forming subunit Kir6.x (Kir6.1 or Kir6.2) and the regulatory subunit sulfonylurea receptor SUR (SUR1 or SUR2). Kir6.x belongs to the inward rectifier K(+) channel family; SUR belongs to the ATP-binding cassette protein superfamily. Heterologous expression of differing combinations of Kir6.1 or Kir6.2 and SUR1 or SUR2 variant (SUR2A or SUR2B) reconstitute different types of K(ATP) channels with distinct electrophysiological properties and nucleotide and pharmacological sensitivities corresponding to the various K(ATP) channels in native tissues. The physiological and pathophysiological roles of K(ATP) channels have been studied primarily using K(ATP) channel blockers and K(+) channel openers, but there is no direct evidence on the role of the K(ATP) channels in many important cellular responses. In addition to the analyses of naturally occurring mutations of the genes in humans, determination of the phenotypes of mice generated by genetic manipulation has been successful in clarifying the function of various gene products. Recently, various genetically engineered mice, including mice lacking K(ATP) channels (knockout mice) and mice expressing various mutant K(ATP) channels (transgenic mice), have been generated. In this review, we focus on the physiological and pathophysiological roles of K(ATP) channels learned from genetic manipulation of mice and naturally occurring mutations in humans. PMID- 12565700 TI - The chondrocyte. AB - The chondrocyte is the resident cell of cartilage that is a prominent tissue in the embryo acting as a template for the development of skeletal elements. In the adult, the distribution of permanent cartilage is much more restricted and is necessary for mechanical support, growth and movement. The cell is isolated within a voluminous extracellular matrix (ECM) that is neither vascularised nor innervated. As a result, nutrient/waste exchange occurs through diffusion and, consequently, under normal and pathological conditions, the cell is unique in its ability to exist in a low oxygen tension environment. Partly as a result of these properties, the tissue has a low reparative potential that, in the case of articular cartilage, predisposes the tissue to degenerative conditions such as arthritis that is a significant clinical problem. Cellfacts. Cytoplasmically isolated. High matrix/cell volume ratio. Do not divide after skeletal maturity unless during pathology. Major contributor to growth of the body. Most energy requirements obtained through glycolysis. PMID- 12565701 TI - Cancer cachexia: the molecular mechanisms. AB - Cancer cachexia is a syndrome characterised by a marked weight loss, anorexia, asthenia and anaemia. In fact, many patients who die with advanced cancer suffer from cancer cachexia. The cachectic state is invariably associated with the presence and growth of the tumour and leads to a malnutrition status due to the induction of anorexia or decreased food intake. In addition, the competition for nutrients between the tumour and the host leads to an accelerated starvation state which promotes severe metabolic disturbances in the host, including hypermetabolism which leads to an increased energetic inefficiency. Although, the search for the cachectic factor(s) started a long time ago, and although many scientific and economic efforts have been devoted to its discovery, we are still a long way from knowing the whole truth. The main aim of the present review is to summarise and evaluate the different catabolic mediators (both humoural and tumoural) involved in cancer cachexia since they may represent targets for future promising clinical investigations. PMID- 12565702 TI - Targeting G protein-coupled A2a adenosine receptors to engineer inflammation in vivo. AB - G protein-coupled adenosine receptors are the subject of intense study as immunomodulators of inflammation especially since the recent demonstration that the A2a receptor acts to down-regulate inflammation and inhibit tissue damage in vivo [Nature 414 (6866) (2001) 916]. The adverse effects of overactive inflammation are evident in diseases e.g. sepsis, rheumatoid arthritis, and multiple sclerosis underscoring the importance of inhibiting inflammation or selectively enhancing inflammatory processes. It has been shown recently that the A2a adenosine receptor is a critical component of an endogenous "immunosuppressive loop" in which extracellular adenosine that accumulates due to local hypoxia caused by inflammatory insult signals through cAMP-elevating A2a receptors in a delayed negative feedback manner. Understanding how tissues regulate inflammation will provide the information necessary to allow for the engineering, or selective targeting, of endogenous inflammatory pathways. Recognition of A2a receptors as "natural" or endogenous brakes of inflammation provides the intellectual scaffolding needed to pursue these goals. PMID- 12565703 TI - Effects of glutathione on kinetics and structural properties of amphibian BbGSTP1 1. AB - Effects of glutathione on the kinetics and structural properties of BbGSTP1-1 were investigated. The liganded state BbGSTP1-1 acquires the capacity to bind the hydrophobic molecules more avidly. Thus, GSH-binding produces significant conformational changes on BbGSTP1-1 which are transmitted to the hydrophobic binding site. Fluorescent experiments carried out with glutathione-analog S methylglutathione suggest that the -SH group of tripeptide is essential for triggering protein conformational changes. It is argued that the capacity of BbGSTP1-1 to be modulated by GSH concentration allows it to play an efficient detoxication action in both aquatic and terrestrial environments. PMID- 12565704 TI - Molecular and biochemical events during differentiation of the HD3 chicken erythroblastic cell line. AB - The chicken erythroblast cell line HD3 is transformed by a temperature-sensitive mutant of avian erythroleukemia virus. Upon shift to the non-permissive temperature in the presence of inducers (hemin and butyric acid), HD3 cells differentiate to an erythrocyte phenotype and provide a model system for analyzing events associated with this process. Expression of some cell surface proteins undergoes drastic changes as cells mature to the erythrocyte stage with a selective loss of membrane proteins that appears to be species-specific. Specific changes also occur in the expression and activities of cytosolic enzymes reflecting alterations of metabolism. HD3 differentiation is characterized by increased transferrin receptor (TFR) expression and increased hemoglobin (Hb) synthesis, a marker for the erythrocyte. In parallel, there is a decrease in glucose transport and an increase in nucleoside transport signifying a switch from glycolytic hexose metabolism to metabolism of pentose from nucleoside. Likewise the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAD) declines while glucose-6-phosphate dehydrogenase (G6PDH) activity remains constant. Commitment to the erythrocyte lineage alters expression of specific genes: TFR mRNA level increases while expression decreases for GLUT1 and GLUT3 glucose transporter mRNAs and GAD mRNA. However, the relationship between GAD activity and GAD mRNA was complex indicating modulation of GAD mRNA and protein half-lives. Serine/threonine and tyrosine phosphorylation and cAMP levels were shown to regulate the level of these messages. In this review, we describe how HD3 differentiation involves changes in plasma membrane composition, metabolism and gene expression that are orchestrated at different levels of control by multiple signaling modalities. PMID- 12565705 TI - Involvement of DNA-dependent protein kinase in down-regulation of cell cycle progression. AB - The catalytic polypeptide of DNA-dependent protein kinase (p470) is encoded by the gene responsible for murine severe combined immunodeficiency (SCID) devoid of DNA double-strand break repair and V(D)J recombination. Here, we have characterized the role of p470 in cell proliferation using SCID mice and the cell lines. In accord with DNA histogram patterns, SCID cell lines (SD/SD-eA and SC3VA2) expressing extremely low level of DNA-PK activity grew faster than a normal mouse cell line (CB/CB-eB) and SC3VA2 complemented with human p470 gene (RD13B2). In regenerating liver after partial hepatectomy, de novo DNA synthesis determined by [(3)H]thymidine incorporation started at 30h in C.B-17/Icr-SCID (SCID) mice and at around 36h in C.B-17/Icr (C.B-17) mice. Compared with normal cells, SCID cells contained slightly higher levels of transcripts of cyclin A, cyclin E, B-Myb and dihydrofolate reductase, which are regulated by E2F-1. E2F-1 playing a key role in G1- to S-phase progression was phosphorylated in vitro by DNA-PK. Importantly, the E2F-1 promoter transcriptional activity in SCID cell lines (SD/SD-eA and SC3VA2) was 4-5-fold higher than that in CB/CB-eB and RD13B2. These results suggest that p470 is involved in down-regulation of cell cycle progression through E2F-1-responsible genes. PMID- 12565706 TI - Scavenger receptor class B, type I on non-malignant and malignant human epithelial cells mediates cholesteryl ester-uptake from high density lipoproteins. AB - Hepatoma cell lines serve as a suitable model to study hepatic clearance of lipoprotein-associated cholesteryl esters (CEs). The present study aimed at investigating holoparticle-association of and selective CE-uptake from human high density lipoprotein subclass 3 (HDL3) by non-malignant adult (Chang-liver) and non-malignant fetal (WRL-68) epithelial cell lines as well as a hepatocellular carcinoma (HUH-7) cell line. Binding properties of 125I-HDL3 at 4 and 37 degrees C were similar for all three cell lines while degradation rates were highest for Chang-liver cells. Calculating the selective uptake of HDL3-associated CEs as the difference between [3H]CE- and 125I-HDL3 cell-association revealed that the selective lipid uptake and holoparticle-association was similar in Chang-liver while in WRL-68 and HUH-7 cells pronounced capacity for lipid tracer uptake in excess of holoparticle uptake was measured. Using RT-PCR, Northern and Western blot analysis, as well as immunocytochemical technique pronounced expression of scavenger receptor class B, type I (SR-BI) but not SR-BII (a splice variant of SR BI less efficient for selective CE-uptake than SR-BI) could be identified in HUH 7 and WRL-68 cells. A polyclonal antiserum raised against SR-BI significantly decreased cell-association of [3H]CE-HDL3 in HUH-7 and WRL-68. The present findings suggest that the capacity for selective cholesteryl ester-uptake from high density lipoprotein by malignant and normal epithelial cells from the liver depends on expression of the scavenger receptor class B, type I. PMID- 12565707 TI - Simultaneous existence of cinnamomin (a type II RIP) and small amount of its free A- and B-chain in mature seeds of camphor tree. AB - Cinnamomin, a type II ribosome-inactivating protein (RIP), was isolated from the mature seeds of camphor tree (Cinnamomum camphora). In this paper, small amount of free A- and B-chain of cinnamomin were found to be present in the mature seed cell of C. camphora besides the intact cinnamomin. Our results demonstrated that camphorin, a type I RIP previously reported to coexist with cinnamomin in the seeds of C. camphora, actually was the A-chain of cinnamomin. The percentage of free A- and B-chain in the total cinnamomin was 2.6-2.8% in the seed extract. Of these free A- and B-chain approximate 80% already existed in the seed cell, only about 20% were produced during the purification operation. As the enzymatic activity to reduce disulfide bond of cinnamomin in the seed extract of C. camphora was detected, we proposed that the free A- and B-chain were derived from the enzymatic reduction of the interchain disulfide bond of cinnamomin. It was demonstrated that the endogenous type II RIPs of several plant species, such as Cinnamomum porrectum, Cinnamomum bodinieri and Ricinus communis, could be enzymatically reduced into the free A- and B-chain in their respective seed cells. The function of the free A-chain in the seed cell and the possibility that metabolic enzymes might be involved in the reduction of the interchain disulfide bond of type II RIPs in vivo are discussed. PMID- 12565708 TI - GM1 ganglioside attenuates convulsions and thiobarbituric acid reactive substances production induced by the intrastriatal injection of methylmalonic acid. AB - The effects of the administration of monosialoganglioside (GM1) on methylmalonic acid (MMA)-induced convulsions, production of thiobarbituric acid reactive substances (TBARS) and on the striatal content of ascorbic acid and total non protein thiol (SH) groups were evaluated in adult male rats. Animals received two intraperitoneal injections of GM1 (50 mg/kg) or saline (0.85% NaCl) spaced 24h apart. Thirty minutes after the second GM1 or saline injection, L-MMA (6 micromol) or NaCl (9 micromol) was injected into the right striatum and the animals were observed for the appearance of convulsions for 15 min. The animals were sacrificed and their striatal content of ascorbic acid, SH groups and TBARS was measured. The effect of GM1 on MMA-induced TBARS production in striatal homogenates was also evaluated in vitro.MMA injection caused convulsions (Sal MMA: 9.8+/-1.4 episodes, which lasted 271+/-48 s) and increased the striatal content of TBARS (Sal-MMA: 149.0+/-11.5 nmol MDA/g tissue), but did not alter total striatal SH or ascorbic acid contents. GM1 pretreatment decreased MMA induced convulsions (GM1-MMA: 6.3+/-2.0 episodes, which lasted 115.1+/-42.2s) and TBARS increase (GM1-MMA: 102.4+/-19.5 nmol MDA/g tissue). GM1 pretreatment increased ascorbic acid content of the striata (saline-pretreated: 1514+/-75.9; GM1-pretreated: 1878.6+/-102.8 microg ascorbic acid/mg tissue). MMA increased TBARS production in vitro, and GM1 had no effect on such MMA-induced effect. This study provides evidence that GM1 increases striatal ascorbic acid content and decreases MMA-induced neurotoxicity assessed by behavioral and neurochemical parameters. PMID- 12565709 TI - Purification and some properties of cathepsin H from rabbit skeletal muscle. AB - Rabbit muscle cathepsin H classified as an aminoendopeptidase was purified and its properties were investigated to clarify its contribution to the proteolysis of postmortem muscle. The purification was performed by ammonium sulfate fractionation and successive chromatographies on Sephadex G-75, phosphocelluose, DEAE-Sephadex A-50 and Sephadex G-100. The purified enzyme gave a single protein band on SDS-PAGE. Its molecular mass was found to be 28 kDa by gel permeation and 30 kDa by SDS-PAGE. The optimum pHs for alpha-N-benzoyl-DL-arginine-beta naphthylamide (BANA)- and L-leucine-beta-naphthylamide (Leu-NA)-hydrolyzing activities were 6.6 and 7.0, respectively. This enzyme was almost stable in the range of pH 4-5 and up to 50 degrees C at pH 5.0. The Km values of BANA- and Leu NA-hydrolyzing activities were 0.367 and 0.203 mM, respectively. The enzyme was inhibited by monoiodoacetic acid, antipain, leupeptin, TLCK and TPCK, but was not affected by pepstatin, bestatin, puromycin, PMSF or trypsin inhibitor. This enzyme strongly acted on Arg-, Lys-, Met-, Ala-, Ser- and Leu-NAs, weakly acted on Val- and Glu-NAs, and hardly acted on Pro- and Gly-NAs. The amount of cathepsin H in muscle was estimated to be less than one-fourth of the sum of the amount of aminopeptidases C and H by the Leu-NA-hydrolyzing activity on the chromatography. This enzyme degraded myosin heavy chain, actin, tropomyosin and troponin I clearly at pH 4.0, while it slightly degraded troponin I at pH 5.0 5.6. Therefore, the contribution of cathepsin H to the proteolysis of postmortem muscle is presumed to be relatively small. PMID- 12565710 TI - Iron overload enhances the development of experimental liver cirrhosis in mice. AB - The role of iron in initiating liver fibrosis in iron overload diseases is not clearly established. Partly, this is due to the lack of suitable animal models that can produce the full liver pathology seen in genetic hemochromatosis. Recent advances in this field have demonstrated that iron may be interacting with other potential liver-damaging agents. The aim of this study was to investigate if feeding with carbonyl iron (CI) facilitates the development of carbon tetrachloride (CCl4)-induced liver fibrosis in the mouse. Mice were given a diet containing 3% CI and treated with CCl4 intraperitoneally twice weekly and 5% alcohol added to the drinking water for 12 weeks. Hepatic iron content increased 15- and 22-fold in animals receiving CI and CI + CCl4. At histological examination, iron-laden hepatocytes were found in CI treated animals, whereas these were absent in animals not exposed to CI. Mice receiving iron-enriched diet alone showed a mild fibrosis. Conversely, a marked collagen deposition was observed in CCl4 and CI + CCl4 groups. In particular, in this latter group, there was evidence of liver cirrhosis. Biochemical evaluation of collagen content substantiated histologic analysis. These results demonstrate that the addition of iron facilitates the development of cirrhosis in animals exposed to subtoxic doses of CCl4. This model may be useful in exploring the pathogenesis of liver cirrhosis. Moreover, its use in genetically altered mouse strains might provide new insight on the role of iron in fibrosis. PMID- 12565711 TI - Transcriptional regulation of the ornithine decarboxylase gene by c-Myc/Max/Mad network and retinoblastoma protein interacting with c-Myc. AB - c-Myc is an oncogenic transcription factor involved in the regulation of cell proliferation, differentiation and apoptosis. The direct targets of c-Myc mediating these various processes are slowly being unravelled. This study indicates that the ornithine decarboxylase (ODC) gene is a physiological transcriptional target of c-Myc in association with induction of cell proliferation and transformation, but not with induction of apoptosis. In addition to the two conserved CACGTG c-Myc-binding sites in the first intron, the CATGTG motif in the 5'-flanking region of the murine odc is also shown to be a functional c-Myc response element. odc is thus a c-Myc target with three binding sites a distance apart. Transient transfection studies with different c-Myc, Max and Mad constructs in COS-7 cells showed that the balance between c-Myc/Max, Max/Max and Max/Mad complexes is crucial for the regulation, resulting in either transactivation or transrepression of an ODC-CAT reporter gene. Transcription of both ODC-CAT and endogenous odc was strongly induced in HeLa cells expressing tetracycline-regulated c-Myc, concomitant with c-Myc promoting the S-phase entry of the cells. Transformation of NIH3T3 cells by c-Ha-ras-(Val12) oncogene was reversed by expression of transcriptionally inactive c-Myc, which was associated with repression of ODC-CAT expression. Further, the c-Myc-induced transactivation of ODC-CAT in COS-7 cells was suppressed by co-expression of the retinoblastoma tumor suppresser pRb, evidently as a result of pRb directly or indirectly interacting with c-Myc. Importantly, the endogenous c-Myc and pRb proteins were also found to associate in Colo 320HSR cells under physiological conditions. These results suggest that c-Myc and pRb can interact in vivo, and may in part control some aspects of cell proliferation and transformation through modulation of odc expression. PMID- 12565712 TI - Increased metabolism of bone collagen in post-menopausal female osteoporotic femoral heads. AB - Our previous studies demonstrated that the residual collagen in osteoporotic bone was not normal but possessed higher levels of lysine hydroxylation and modified cross-linking. However, the mechanism for these changes was not clear. In the current investigation, an assessment of bone collagen metabolism in osteoporosis (OP) revealed an increase in the overall metabolism of collagen relative to age matched controls. The increased metabolism accounts for the observed post translational modifications of collagen which lead to a more fragile bone matrix. The rate of bone metabolism is therefore an important aspect of the pathogenesis of osteoporosis, the greater the turnover the greater the propensity of a more fragile tissue. Clearly, the quality of bone tissue does not depend solely on adequate bone density but also on the state of the collagenous matrix. PMID- 12565713 TI - Charge-based heterogeneity of human plasma lipoproteins at hypertriglyceridemia: capillary isotachophoresis study. AB - To reveal the metabolic links between and within pools of pro-atherogenic triglyceride(TG)-rich lipoproteins and anti-atherogenic high density lipoproteins (HDL), the changes in lipoprotein profile at hypertriglyceridemia were analyzed by capillary isotachophoresis. Plasma samples from patients with apoE3/3 phenotype were stained with a fluorescent probe NBD-C6-ceramide and lipoproteins resolved into six H-, one (V+I) and four L-components which belong to HDL, very low and intermediate density (VLDL+IDL) and low density lipoproteins (LDL), respectively. The expected correlation between the relative size of the combined fractions and lipid and apolipoprotein values was obtained confirming the validity of the approach. The new findings were obtained as follows. (1) The fast L-component correlated inversely with HDL-cholesterol (Chol), while intermediate and slow H-components correlated inversely with plasma and LDL-Chol and apoB. (2) The content of intermediate and slow H-components increased within H-pool and decreased relative TG-rich lipoproteins as hypertriglyceridemia rose due to the impairment of triglyceride hydrolysis by lipoprotein lipase within TG-rich particles. (3) A predictive value of the ratios of fast to slow H-components as an indicator of lecithin:cholesterol acyltransferase activity was demonstrated which tended to decrease at hypertriglyceridemia. (4) The L1/L2 ratio may be considered as an indicator of the accumulation of small dense LDL, which is a feature of clinically manifested atherogenic B-pattern. The competition between H(DL) and L(DL) particles for hepatic lipase and significant contribution of apoE to functional deficiency of H(DL) particles at hypertriglyceridemia are suggested. PMID- 12565714 TI - The performance of two rapid quantitative D-dimer assays in 287 patients with clinically suspected pulmonary embolism. AB - CONTEXT: Objective tests are necessary for the diagnosis of pulmonary embolism (PE). D-dimer assays have been suggested as useful screening tests to exclude this diagnosis. OBJECTIVE: To compare the diagnostic accuracy of two rapid quantitative D-dimers in patients with suspected pulmonary embolism. DESIGN: Plasma D-dimer levels were measured using two commercially available assays (Tinaquant and Vidas). A strict imaging protocol was used to arrive at a final diagnosis of PE or deep venous thrombosis (DVT). SETTING: Multicenter study in six Dutch referral centers. PATIENTS: A total of 287 in- and outpatients with clinically suspected pulmonary embolism. MAIN OUTCOME MEASURES: Diagnostic accuracy indices for the two assays were calculated and additional receiver operated characteristics (ROC) analysis was performed. RESULTS: Using the manufacturer's advised cutoff values, the sensitivity and specificity were 88% and 52% for Vidas and 82% and 61% for Tinaquant, respectively. These differences were statistically significant (McNemar, P < 0.0001). However, no statistical differences were found between the two assays using ROC analysis (AUC = 0.78 for both assays). CONCLUSIONS: Both quantitative D-dimer tests had similar diagnostic accuracy; however, at the manufacturer's advised cutoff level, Vidas performed significantly better. Nevertheless, to safely exclude pulmonary embolism, D-dimer assays should be combined with other diagnostic tests. PMID- 12565715 TI - Low-dose warfarin in rehabilitating stroke survivors. AB - BACKGROUND: Patients undergoing rehabilitation after thromboembolic stroke have a relatively high incidence of venous thromboembolism (VTE). Warfarin, with a target international normalized ratio (INR) of 2.0-3.0 is effective for the prevention of VTE. However, because stroke is a major risk factor for bleeding with warfarin, a less intense regimen (target INR < 2.0), might safely prevent VTE in stroke rehabilitation patients. METHODS: This study was a randomized, double-blind, placebo-controlled trial of 2 mg of warfarin in patients undergoing rehabilitation following completed stroke. The major efficacy endpoint was symptomatic, objectively proven VTE or asymptomatic VTE detected by monthly duplex ultrasonography (DU) of the proximal leg veins or mandatory bilateral contrast venography performed at the end of the study. The major safety endpoint was bleeding. RESULTS: There were 475 patients screened for enrollment, 355 had one or more exclusion criterion, and 17 had previously undetected proximal DVT on admission. Of the 103 eligible and consenting patients, 56 received warfarin and 47 received placebo. Of the randomized patients, 88 had successful venography (47 warfarin and 41 placebo). In the warfarin group, three (8%) patients had DVT and one (2%) had proximal DVT whereas in the placebo group, seven (20%) had DVT and five (13%) had proximal DVT. The risk ratio for any DVT in warfarin-treated patients relative to placebo-treated patients was 0.39 (95% confidence interval (CI), 0.13-1.37). For proximal DVT, the risk ratio was 0.17 (95% CI, 0.01-1.4). No patients suffered major bleeding. CONCLUSIONS: A fixed dose of 2 mg of warfarin per day in patients undergoing stroke rehabilitation is safe and associated with a relative risk reduction of about 80% in the incidence of proximal DVT and 60% in overall DVT. PMID- 12565716 TI - Epitope specificity of anti-FVIII antibodies during immune tolerance therapy with factor VIII preparation containing von Willebrand factor. AB - The study aimed at characterizing the putative changes in the epitope specificity of anti-FVIII antibodies during a successful immune tolerance treatment of the haemophilia A patient with the factor VIII (FVIII) preparation containing the von Willebrand factor (VWF). At the beginning of treatment, anti-FVIII inhibitory antibodies recognizing predominantly the light chain of FVIII were prevalent and persisted throughout the treatment. More detailed characterization of the FVIII antibody epitope specificity by using GST-fusion proteins corresponding to different FVIII domains revealed the prevalence of C1-domain-specific antibodies, while a remarkably lower amount of antibodies were targeted at the C2 and the a3 domains of the FVIII light chain and towards the A2 and the A1 domain of the FVIII heavy chain. The epitope specificity of antibodies remained rather unchanged throughout treatment except the elevated level of C2-domain-specific FVIII antibodies after a temporary interruption of treatment. The patient's antibodies were unable to interfere with the FVIII binding to VWF or to phospholipids, but inhibited FXa generation and the binding of FX to FVIII on the phospholipid monolayer. Thus, a unique pattern of the epitope specificity of FVIII antibodies and the mechanism to inhibit FVIII:C activity by FVIII-light chain-specific antibodies were characterized. PMID- 12565717 TI - Effects of glycoprotein IIb/IIIa antagonists on platelet activation: development of a transfer method to mimic peak to trough receptor occupancy. AB - Several oral glycoprotein (GP) IIb/IIIa antagonists, Sibrafiban, Orbofiban and Lotrafiban, have been studied in large phase III trials; each has failed to provide efficacy and has been associated with increased mortality. Roxifiban has pharmacokinetic and pharmacodynamic properties believed to be more favorable than the earlier oral agents. Here, we revisit the controversial hypothesis of platelet activation liabilities of GP IIb/IIIa antagonists. The effects of site occupancy by four fibans (Roxifiban, Sibrafiban, Orbofiban and Lotrafiban) on platelet activation was assessed using P-selectin expression, fibrinogen binding and microaggregate formation. All four fibans inhibited ADP and TRAP-stimulated fibrinogen binding and microaggregate formation in a concentration-dependent manner, whereas P-selectin expression was relatively unaltered. To more vigorously test for activation liabilities, the effects of transition from peak to trough receptor occupancy upon platelet stimulation was analyzed. The high affinity of Roxifiban for resting platelets precluded reduction of site occupancy by dialysis or gel filtration. A method was developed that takes advantage of the rapid equilibrium of Roxifiban between platelets and soluble GPIIb/IIIa. The platelet occupancy is controlled by the ratio of platelet GPIIb/IIIa to soluble GPIIb/IIIa. This method allows in vitro investigation of peak/trough transitions on platelet activation. A decrease in site occupancy from peak to trough of Roxifiban or Sibrafiban did not result in increased activation of platelets. The loss of platelet-bound antagonist upon incubation with purified soluble GPIIb/IIIa returned fibrinogen binding/microaggregate formation to no drug levels. In conclusion, these studies do not provide evidence for an activation liability of GPIIb/IIIa antagonists in vitro. PMID- 12565718 TI - Enhancing effect of advanced glycation end products on serotonin-induced platelet aggregation in patients with diabetes mellitus. AB - Advanced glycation end products (AGEs) are thought to be responsible for some complications of diabetes mellitus (DM), including microangiopathy. Plasma serotonin is increased in diabetes mellitus patients, and this increase is related, at least in part, to platelet hyperfunction. In order to clarify the relationship between advanced glycation end products, serotonin, and thrombotic complications in diabetes mellitus patients, we examined the effect of advanced glycation end products on serotonin-induced platelet aggregation. In diabetic patients, although serotonin-induced platelet aggregation was enhanced with an increase in serum-advanced glycation end products, there was no correlation between platelet aggregation and either hemoglobin A1c or fasting blood sugar. To examine the direct effect of advanced glycation end products on platelet aggregation, we prepared advanced glycation end products by in vitro incubation of human albumin with glucose (250 mM) at 37 degrees C for 8 weeks. Serotonin induced platelet aggregation was dose-dependently increased by advanced glycation end products. Adenosine diphosphate-induced platelet aggregation also was increased by advanced glycation end products, but this increment was diminished by addition of sarpogrelate, a selective serotonin receptor antagonist. These results suggest that advanced glycation end products enhance platelet aggregation through the serotonin receptor, and perhaps influencing the development of thrombotic complications in diabetic patients. PMID- 12565719 TI - Enhanced platelet aggregation with TRAP-6 and collagen in platelet aggregometry in patients with venous thromboembolism. AB - The role of platelet hyperaggregability as a possible risk factor for venous thromboembolism is not well defined. Some authors described enhanced maximal platelet aggregation in platelet aggregometry as a contributing factor for arterial and venous thrombosis. This syndrome has been termed "sticky-platelet syndrome" (SPS). The diagnosis of SPS is based on the demonstration of platelet hyperaggregability in aggregometry after stimulation with epinephrine (EPI) and/or adenosine diphosphate (ADP). We investigated platelet hyperaggregability in platelet-rich plasma (PRP) of patients (n = 34) with unexplained venous thromboembolism in comparison to healthy individuals (n = 53). For analysis, platelet aggregometry was performed and the influence of epinephrine, adenosine diphosphate, collagen (Coll) and thrombin receptor-activated peptide (TRAP-6) as agonist were determined. Compared to the control group, patients with venous thromboembolism showed an enhanced maximal platelet aggregation with low concentrations of TRAP-6 (2 microM) and collagen (0.05 microM). In contrast, we could not detect an increased platelet aggregation with EPI or ADP. Our results indicate that platelet hyperaggregability may represent an independent risk factor in patients with otherwise unexplained venous thromboembolism. In our study, low concentrations of TRAP-6 and collagen are superior to EPI and ADP to define platelet hyperreactivity in platelet aggregometry. PMID- 12565720 TI - The hemostatic agent ethamsylate enhances P-selectin membrane expression in human platelets and cultured endothelial cells. AB - Ethamsylate possesses antihemorrhagic properties, but whether or not it directly activates blood platelets is unclear. Here we investigated the platelet activation potential of ethamsylate, by measuring membrane P-selectin expression with flow cytometry in human whole blood and also by immunofluorescence imaging of isolated human platelets. Moreover, we measured membrane P-selectin expression in the SV40-transformed aortic rat endothelial cell line (SVAREC) and 14C ethamsylate membrane binding and/or uptake in platelets and endothelial cells. Whole blood flow cytometry showed a modest, but statistically significant increase by ethamsylate in the percentage of platelets expressing P-selectin (from 2% to 4-5%, p < 0.05). Immunofluorescence showed a sizable (39%) and significant (p < 0.01) enhancement of P-selectin expression at the lowest concentration of ethamsylate tested (1 microM), with maximal enhancement of P selectin expression (75-90%) at 10 microM ethamsylate. Similar results were obtained in SVAREC endothelial cells. 14C-ethamsylate specifically bound to platelets and endothelial cell membranes, without significant uptake into the cell interior. In conclusion, ethamsylate enhances membrane P-selectin expression in human platelets and in cultured endothelial cells. Ethamsylate specifically binds to some protein receptor in platelet and endothelial cell membranes, receptor which can signal for membrane P-selectin expression. These results support the view that ethamsylate acts on the first step of hemostasis, by improving platelet adhesiveness and restoring capillary resistance. PMID- 12565721 TI - Vascular dementia in patients with immune thrombocytopenic purpura. AB - INTRODUCTION: Platelets have been implicated in memory disorders but this has not been investigated in patients with immune or idiopathic thrombocytopenia (ITP). ITP is an autoimmune disorder in which autoantibodies bring about platelet destruction. We previously reported a group of ITP patients who manifested TIA like syndrome and gradual memory loss leading to dementia: platelet microparticles (PMP), a marker of platelet activation, were often elevated, suggesting that procoagulant PMP released from stimulated platelets contributed to thrombosis in small vessels. We have expanded on those studies to better define the clinical, laboratory, and radiologic characteristics of this syndrome. MATERIALS AND METHODS: Twenty ITP patients with this syndrome were studied in comparison to twenty-three ITP patients without it (patient controls). Clinical and laboratory features were compared and radiologic images were analyzed. Factors influencing the rate of progression to advanced dementia were also investigated. RESULTS AND CONCLUSION: Recurring dizzy or weak spells, TIA-like syndrome, recent memory loss, and cognitive impairment were common initial complaints. In some, these symptoms progressed rapidly to dementia but was indolent in others. Progression was faster in those with splenectomy and higher platelet counts. MRI showed enhanced signal in subcortical, periventricular areas, consistent with ischemic small vessel disease. Compared to patient controls, bleeding was less frequent and platelet activation (increased PMP, CD62p) was more frequent in the study group. Thrombotic complications may occur in ITP, manifested as TIA-like syndrome or memory loss due to ischemic small vessel disease, progressing to vascular dementia. Memory disturbances associated with platelet disorders warrants further investigation. PMID- 12565722 TI - Weak platelet agonists and U46619 induce apoptosis-like events in platelets, in the absence of phosphatidylserine exposure. AB - Platelets express apoptotic markers during storage, while aging and after stimulation with strong agonists thrombin and collagen. It is unknown if the weak agonists ADP and epinephrine or U46619, a thromboxane analog, induce the expression of apoptotic markers in platelets. To answer this question, we measured phosphatidylserine exposure, gelsolin cleavage and decrease in membrane mitochondrial potential after stimulation with these agonists. No phosphatidylserine exposure was evident, however, gelsolin cleavage and a platelet population with a decreased membrane mitochondrial potential appeared, suggesting that in platelets selective agonists can induce apoptosis in the absence of phosphatidylserine exposure. Interestingly, costimulation by thrombin plus collagen together with each of the other agonists increased the phosphatidylserine exposure induced by strong agonists. These findings may be of importance in platelet activation and apoptosis under pathophysiological conditions where multiple effectors are involved. PMID- 12565723 TI - Effects of activated factor X inhibitor, JTV-803, in a pig model of hemodialysis. AB - JTV-803 (4-[(2-amidino-1,2,3,4-tetrahydroisoquinolin-7-yloxy)methyl]-1-(4 pyridinyl)piperidine-4-carboxylic acid monomethanesulfonate trihydrate), a specific inhibitor of factor Xa, was evaluated in a pig hemodialysis model with ligation of renal arteries. In this model, JTV-803 administered into the dialysis circuit showed an anticoagulant effect (prolongation of dialysis time) at doses of 0.3 mg/kg + 0.3, 1.0 and 3.0 mg/kg/h. The prolongation of dialysis time in the 0.3 mg/kg + 1.0 mg/kg/h JTV-803 group was comparable to that in the 15 U/kg + 7.5 U/kg/h heparin group. The plasma concentrations of JTV-803 following 0.3 mg/kg + 1.0 mg/kg/h infusion of the drug into the dialysis circuit was 500 ng/ml or less, and prothrombin time was 1.5-fold or less than the pretreatment value. JTV-803 was removed by passing the blood through a dialyzer, resulting in a clearance rate of 53.4-81.8%. After the end of dialysis, plasma concentrations of JTV-803 decreased rapidly with time. These results suggest that human factor Xa inhibitor JTV-803 may have good potential as an antithrombotic agent during hemodialysis, with lower likelihood of bleeding after the end of dialysis. PMID- 12565724 TI - Effect of some new thioglycosides on endotoxin-induced disseminated intravascular coagulation in rabbits. AB - Disseminated intravascular coagulation (DIC) is a systemic thrombohemorrhagic disorder seen in association with many clinical situations, e.g. sepsis, malignancy, obstetrical complications and intravascular hemolysis. In our model, disseminated intravascular coagulation was induced in rabbits by two consecutive intravenous bolus injections of endotoxin from Escherichia coli, 80 and 40 microg/kg. The control group was treated with 0.9% saline. The activity of thioglycosides was compared to unfractionated heparin (UFH) and efegatran with and without administration of endotoxin. Drugs were administered in the following doses: heparin 50 and 100 IU/kg/h i.v. infusion; efegatran 0.25 and 0.5 mg/kg/h i.v. infusion; GYKI 39521 (RGH-1875) as well as GYKI 39541 (RGH-1962) 12.5 and 25 mg/kg per os. Thioglycosides did not modify coagulation parameters in this model [prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT)] as compared with endotoxin/vehicle group. The changes in TFPI level after administration of thioglycosides and heparin were similar in the mentioned model to those without endotoxin. Endotoxin-induced changes of leukocyte count were not affected by GYKI 39521 and GYKI 39541 treatment in our model. Diminution of fibrinogen level and platelet count was prevented by GYKI 39521 and GYKI 39541. Fibrin degradation products and fibrinolysis were significantly decreased by GYKI 39521 and GYKI 39541. The thioglycosides may have a lower risk of bleeding in the treatment of disseminated intravascular coagulation than heparin. PMID- 12565725 TI - In vitro effect of a thrombin inhibition peptide selected by phage display technology. AB - A repeated selection of phages from a cyclic heptapeptide phage display library resulted in the enrichment of phages that bind to human alpha-thrombin. One clone of the binding phages that competed with PPACK for binding to thrombin and that had the best binding characteristics was chosen. The amino acid sequence of the peptide displayed on this phage was determined and a peptide with the sequence, Cys-Asn-Arg-Pro-Phe-Ile-Pro-Thr-Cys was synthesised. This peptide, thrombin inhibiting peptide (TIP), is a full competitive inhibitor of thrombin with an inhibition constant (K(i)) of 0.4974 mM. It lengthened the thrombin time and inhibited thrombin-induced platelet activation and the platelet release reaction, both in a dose-dependent manner. It also reduced platelet adhesion onto a human microvascular endothelial matrix in the parallel plate flow chamber under both arterial and venous shear conditions. Thus, we have selected and synthesised a cyclic heptapeptide that competes with PPACK to bind to thrombin and that can be developed as a direct antithrombin. PMID- 12565726 TI - A web based video tool for MR arthrography. AB - OBJECTIVE: Our objective was to integrate digital video and web technology to construct a useful self paced learning tool to deliver recorded techniques of magnetic resonance (MR) arthrography for junior radiology residents and medical students. CONCLUSION: The streaming of audio media, an emerging technology, is an effective vehicle to deliver educational material over the Internet using commercially available PCs and modems, especially when teaching resources are limited. PMID- 12565727 TI - Automated lung outline reconstruction in ventilation-perfusion scans using principal component analysis techniques. AB - The present work addresses the development of an automated software-based system utilized in order to create an outline reconstruction of lung images from ventilation-perfusion scans for the purpose of diagnosing pulmonary embolism. The proposed diagnostic software procedure would require a standard set of digitized ventilation-perfusion scans in addition to correlated chest X-rays as key components in the identification of an ideal template match used to approximate and reconstruct the outline of the lungs. These reconstructed lung images would then be used to extract the necessary PIOPED-compliant features which would warrant a pulmonary embolism diagnosis. In order to evaluate this issue, two separate principal component analysis (PCA) algorithms were employed independently, including Eigenlungs, which was adapted from the Eigenfaces method, and an artificial neural network. The results obtained through MATLAB(TM) simulation indicated that lung outline reconstruction through the PCA approach carries significant viability. PMID- 12565728 TI - Physiological state model for human ergonomic workload. AB - Twenty ergonomic tasks were evaluated in which human operators performed mixed static work and dynamic work. Steady-state physiological data are the input into a model as regressor variables, which are then multiplied by the respective regressor coefficients. The resultant physiological state model output is a single response variable that represents the workload. Mixed stepping regression techniques were utilized to calculate the regressor coefficients. Ten physiological state model equations resulted. A lower order equation (with three regressor variables) accounted for 80% of the observed variance. The highest order equation (with ten regressor variables) accounted for 89% of the variance. PMID- 12565729 TI - VRML metabolic network visualizer. AB - A successful date collection visualization should satisfy a set of many requirements: unification of diverse data formats, support for serendipity research, support of hierarchical structures, algorithmizability, vast information density, Internet-readiness, and other. Recently, virtual reality has made significant progress in engineering, architectural design, entertainment and communication. We experiment with the possibility of using the immersive abstract three-dimensional visualizations of the metabolic networks. We present the trial Metabolic Network Visualizer software, which produces graphical representation of a metabolic network as a VRML world from a formal description written in a simple SGML-type scripting language. PMID- 12565730 TI - The selectivity of scorpion alpha-toxins for sodium channel subtypes is determined by subtle variations at the interacting surface. PMID- 12565731 TI - Significance of assessment of serum cardiac troponin I and interleukin-8 in scorpion envenomed children. AB - In the present study, 41 children in Upper Egypt were admitted to Pediatric Intensive Care Unit, Assiut University Hospital, for scorpion envenomation. They were compared with 15 apparently healthy children of matching age as controls. The victims and controls were subjected to complete clinical examination, full blood count and arterial blood gases analysis. According to severity of scorpion envenomation, 17 children had manifestations of severe envenomation and clinical signs of toxic myocarditis (severe cases), 14 children had moderate manifestations of envenomation without clinical evidence of carditis (moderate cases) and 10 cases showing only mild symptoms of envenomation (mild cases). The serum levels of cardiac troponin I (cTnI) and interleukin-8 (IL-8) beside the enzymatic activities of creatine phosphokinase (CPK), CPK-isoenzyme-MB (CPK-MB) and lactate dehydrogenase (LDH) were determined once for mild cases and controls on admission and twice for severe and moderate cases on admission and after 24. Electrocardiography and measurements of echocardiographic (Echo) of % fractional shortening of left ventricule (% SF), left ventricular ejection fraction (LVEF) and cardiac chambers dilatation were done for severe and moderate cases. All the envenomed victims showed significantly higher mean values of CPK, CPK-MB, LDH, and IL-8 on admission in comparison to control group. cTnI was not detectable in the sera of control group as well as patients of mild envenomation. The mean values of CPK, CPK-MB, LDH, and IL-8 were significantly higher in severe cases while only IL-8 and CPK-MB were significantly higher in moderate cases in comparison with mild cases. The mean values of IL-8, cTnI, CPK, CPK-MB and LDH were significantly higher in severe cases both on admission and on follow-up comparing with moderate cases. The case fatality rate was 12.5% and all were from severe cases with toxic myocarditis. The non-survivors victims showed significant higher mean values of only cTnI on admission and both cTnI and IL-8 on follow up in comparison to the survivors. Significant reduction of % SF and LVEF were noticed among the non-survivors in comparison to survivors. The cTnI showed 100% specificity and sensitivity for diagnosis of myocardial injury in relation to Echo finding in the envenomed victims. In severe cases, cTnI was positively correlated with IL-8 while negatively correlated with %SF and LVEF. In conclusion, cTnI is a specific marker for diagnosis of myocardial injury in scorpion envenomation while other biochemical markers did not show such specificity. Also, IL-8 may be involved in the pathogenesis of myocardial injury of scorpion envenomation. Both cTnI and IL-8 may be useful to forecast the fatal outcome in scorpion envenomation. PMID- 12565732 TI - Biochemical and biological activities of the venom of a new species of pitviper from Vietnam, Triceratolepidophis sieversorum. AB - Biochemical and biological activities of a venom sample from a recently discovered new genus and species of pitviper from Vietnam, Triceratolepidophis sieversorum, were assayed and compared with those of five other viperid snakes (Bothrops asper, Crotalus atrox, Protobothrops flavoviridis, Trimeresurus insularis, and Vipera ammodytes). The venom had high casein hydrolysis, arginine ester hydrolysis and haemorrhagic activities, lacked measurable phosphodiesterase and L-amino acid oxidase activities, and had no procoagulant activity on either bovine fibrinogen or human plasma. Other enzymatic activities (phospholipase A(2), kallikrein) were moderate. The approximate i.p. LD(50) (mice) of the venom is about 5-6 mg/kg. PMID- 12565733 TI - Ubiquitous 'benign' alga emerges as the cause of shellfish contamination responsible for the human toxic syndrome, azaspiracid poisoning. AB - A new human toxic syndrome, azaspiracid poisoning (AZP), was identified following illness from the consumption of contaminated mussels (Mytilus edulis). To discover the aetiology of AZP, sensitive analytical protocols involving liquid chromatography-mass spectrometry (LC-MS) were used to screen marine phytoplankton for azaspiracids. Collections of single species were prepared by manually separating phytoplankton for LC-MS analysis. A dinoflagellate species of the genus, Protoperidinium, has been identified as the progenitor of azaspiracids. Azaspiracid-1, and its analogues, AZA2 and AZA3, were identified in extracts of 200 cells using electrospray multiple tandem MS. This discovery has significant implications for both human health and the aquaculture industry since this phytoplankton genus was previously considered to be toxicologically benign. The average toxin content was 1.8 fmol of total AZA toxins per cell with AZA1 as the predominant toxin, accounting for 82% of the total. PMID- 12565734 TI - Synthesis and organotropism of 3H-dihydro derivatives of the cyanobacterial peptide hepatotoxin nodularin. AB - Tritium-labelled dihydro derivatives of the cyanobacterial peptide hepatotoxin nodularin were prepared by reduction with sodium boro[3H]hydride. The optimised reaction gave two dihydronodularin stereoisomers which were purified by high performance liquid chromatography with a mobile phase of methanol-0.7% sodium sulfate (6:4) and a C(18) stationary phase. The specific activities of the stereoisomers were 1780-1807 dis min(-1) ng(-1). The radiolabelled dihydronodularins were tested for stability and used for toxicokinetic studies in mice. Liver was the main site of toxin accumulation. PMID- 12565735 TI - BotIT6: a potent depressant insect toxin from Buthus occitanus tunetanus venom. AB - A new depressant insect toxin Buthus occitanus tunetanus insect-toxin 6 (BotIT6) was purified by high-performance liquid chromatography from Buthus occitanus tunetanus (Bot) venom. BotIT6 is very active against Blatella germanica (LD50=10ng/100mg body mass) thus being one of the most potent anti-insect toxin so far characterised. When compared to other insect toxin sequences, BotIT6 present high similarities with depressant insect toxins with an additional arginine residue at the C-terminus and a methionine at position 27. The calculated net charge of BotIT6 is positive (+3) whereas it is negative for classical depressant toxins: this might be associated with its high toxicity. Voltage current clump studies show that BotIT6 is not a very potent depressant insect toxin despite its high toxicity in vivo. BotIT6 is able to fully inhibit the specific binding of 125I AaHIT and 125I-BotIT2 on Periplaneta americana synaptosomal membrane vesicles with high affinities. Despite its higher toxicity BotIT6 is a weaker competitor with 125I AaHIT and 125I BotIT2 as compared to the other beta toxins.Altogether, these results may suggest that BotIT6 probably defines a novel sub-group of depressant anti-insect toxins for which the receptor site can be overlapping, but not identical to that for classical depressant insect toxins. PMID- 12565736 TI - Disulfide bridges and blockage of Shaker B K(+)-channels by another butantoxin peptide purified from the Argentinean scorpion Tityus trivittatus. AB - A peptide was isolated from the venom of the scorpion Tityus trivittatus. It is an isoform of the toxin TsTX-IV earlier described [Toxicon 37 (1999) 651] and identical to butantoxin [Arch. Biochem. Biophys. 379 (2000) 18], both isolated from the Brazilian scorpion Tityus serrulatus. This newly characterized peptide contains 40 amino acid residues with a molecular mass of [M+H(+)] 4507.0, cross linked by four disulfide bridges, made between the cysteine pairs: Cys2-Cys5, Cys10-Cys31, Cys16-Cys36 and Cys20-Cys38. It blocks in a completely reversible manner the Shaker B K(+)-channels, with a K(d) around 660nM. It belongs to the sub-family 12 and it is now being classified as alpha-KTx 12.2. PMID- 12565737 TI - Enrichment of the antibodies against the C-terminus of Taiwan cobra cobrotoxin using dimeric glutaraldehyde-modified toxin as an immunogen. AB - The repertoire of antibodies producing by immunizing rabbits with cobrotoxin and dimeric glutaraldehyde-modified cobrotoxin (dGA-cobrotoxin) was analyzed by studying the immunoreactivity of the two antibody preparations toward cobrotoxin, GA-cobrotoxin and recombinant cobrotoxin. The results of enzyme-linked immunoassay revealed that the two antibody preparations exhibited a higher reactivity against their cognate antigen. Moreover, different behavior was observed for the reactivity of the two antibody preparations against GA cobrotoxin and recombinant cobrotoxin. Notably, distortion of disulfide linkages at the C-terminus resulted in a reduced decrease in the antigenic activity of recombinant cobrotoxin toward anti-cobrotoxin antibodies compared to anti-dGA cobrotoxin antibodies. Affinity purification of the antibodies against the C terminus of cobrotoxin revealed that its amount represented 77% and 35.5% of the total anti-dGA-cobrotoxin antibodies and the total anti-cobrotoxin antibodies, respectively. These findings suggest that the antibody preparation elicited by dGA-cobrotoxin enriches the content of antibodies recognizes the C-terminal region of native cobrotoxin. PMID- 12565738 TI - Seasonality of diarrhetic shellfish poisoning at a coastal lagoon in Portugal: rainfall patterns and folk wisdom. AB - Of the three types of toxicity known so far in Portuguese shellfish, only diarrhetic shellfish poisoning (DSP) and amnesic shellfish poisoning (ASP) are produced by microalgae that seem to have been present in the last decades or centuries. The most important paralytic shellfish poisoning (PSP) producer, Gymnodinium catenatum, is hypothesised to have been introduced quite recently as only in 1976 PSP toxicity was detected for the first time in shellfish from Galicia, NW Iberian Peninsula. While ASP presents very short episodes of contamination, the concentration of DSP toxins in some years surpasses human safety values for much longer periods. It is traditionally stated that shellfish should be consumed in 'months with R' (September-April). A retrospective study of the maximum monthly DSP levels attained in mussels from a coastal lagoon-Ria de Aveiro-between 1994 and 2001, showed that the highest frequency of months with concentrations surpassing the safety level of 2 microg/g digestive glands were found in June-September, followed by May and October. These months correspond with the months of lowest historical average rainfall in the period 1941-1998. Oscillations in the rainfall pattern coincided with earliest (or latest) detection by HPLC of DSP toxins in mussel in the years studied. In a semi-closed lagunar environment prone to in situ growth of DSP-producer microalgae, like Dinophysis acuminata, rainfall affects river output, lowering salinity and disrupting water column stability that favours Dinophysis growth. The seasonality of DSP recurrence may be connected to the folk adage on safety of shellfish consumption, after many years of empirical observations by coastal populations of diarrhoea episodes in summertime. PMID- 12565739 TI - Spatial and temporal evolution of PSP toxins along the Atlantic shore of Morocco. AB - A monitoring program for bivalve molluscs contaminated by algal toxins was established in 1992 at different stations along the Atlantic Moroccan shore. The presence of toxicity in bivalve molluscs commercially exploited was tested fortnightly using the mouse bioassay method. Results obtained from this surveillance indicate paralytic shellfish poisoning is responsible for bivalve molluscs contamination along the Atlantic coastline of Morocco. Toxin profile was established by automated pre-column HPLC/FLD in selected contaminated tissues. The study of individual toxins in mussel during a bloom in the northern Atlantic coastline in 1994 showed a very complex profile, typical to that obtained with cultures of the toxigenic dinoflagellate Gymnodinium catenatum isolated from the Iberian region. However, toxin composition of mussels and marine beans from later blooms in the southern Atlantic coastline in 1999 showed a strong resemblance with that of Alexandrium minutum, due to dominance of gonyautoxins 1/4. A minor contamination by G. catenatum due to the presence of decarbamoyl-saxitoxin is hypothesized. PMID- 12565740 TI - Effects of muscarinic toxins MT2 and MT7, from green mamba venom, on m1, m3 and m5 muscarinic receptors expressed in Chinese Hamster Ovary cells. AB - Several small proteins called muscarinic toxins (MTs) have been isolated from venom of green mamba (Dendroaspis angusticeps). They have previously been shown in radioligand binding studies to have high selectivity and affinity for individual muscarinic receptor subtypes, but less is known of their functional effects. This study has examined the actions of two of these MTs, MT2 and MT7, using changes in cytosolic Ca(2+) ([Ca(2+)](i)) measured using the fluorescent indicator fura-2 in Chinese Hamster Ovary (CHO) cells stably transfected with individual muscarinic receptor subtypes, m1, m3 and m5. MT2 activated the m1 receptor: at concentrations above 100 nM it caused significant and concentration dependent increases in [Ca(2+)](i). From 25 to 800 nM MT2 also produced increases in [Ca(2+)](i) by activating m3 receptors, although these increases in [Ca(2+)](i) were not strictly concentration-dependent with only intermittent responses being recorded (i.e. it was not always possible to obtain a response to the agonist with each application of the compound). MT2 (800-1600 nM) also caused significant increases in [Ca(2+)](i) in CHO cells expressing the m5 muscarinic receptor subtype. MT7 (1 microM) displayed no agonist activity at any of the muscarinic receptors but was a potent non-competitive antagonist (at 20 nM) at the m1 muscarinic receptor subtype. It had no antagonist activity at the m3 or m5 subtypes. These results indicate that MT7 is a highly specific antagonist at the m1 muscarinic receptor subtype as suggested by results from radioligand binding studies. However, MT2 is less selective for the m1 muscarinic receptor than previously described as it also exhibits agonist activity at the m3 and m5 muscarinic receptors, which was not detected in radioligand binding studies. PMID- 12565741 TI - Molecular cloning and expression of structural domains of bothropasin, a P-III metalloproteinase from the venom of Bothrops jararaca. AB - Mature P-III snake metalloproteinases are soluble venom components which belong to the Reprolysin sub family and are structurally related to the mammalian membrane-bound A Disintegrin And Metalloproteinase (ADAMs). Here we present the molecular cloning of bothropasin, a metalloproteinase with hemorrhagic and myonecrotic activities isolated from the venom of Bothrops jararaca. The full length cDNA encoding the bothropasin precursor was cloned by immunoscreening and its authenticity was confirmed by the amino acid sequence of internal fragments obtained from an autolyzed sample of native bothropasin. The predicted bothropasin precursor is comprised of the elements of a P-III venom metalloproteinase: signal sequence, pro-, metalloproteinase, disintegrin-like and cysteine-rich domains. In the autolysis process of native bothropasin, the disintegrin-like and cysteine-rich domains remained intact while the metalloproteinase domain was cleaved at different sites. The attempts made to obtain the recombinant precursor form of bothropasin using bacterial, yeast and mammalian cell expression systems failed to produce it in an amount sufficient to analyze the activation of the zymogen. Nevertheless, the study of the expression of the individual domains of bothropasin using a bacterial system resulted in the production of recombinant pro-and disintegrin-like+cysteine-rich domains but not the metalloproteinase domain. These results along with the autolysis pattern of the native protein suggest a role for the metalloproteinase domain in the structural stability of bothropasin. PMID- 12565742 TI - An epidermal growth factor-like toxin and two sodium channel toxins from the sea anemone Stichodactyla gigantea. AB - Three peptide toxins (gigantoxins I-III) with crab toxicity were isolated from the sea anemone Stichodactyla gigantea by gel filtration on Sephadex G-50 and reverse-phase HPLC on TSKgel ODS-120T and their complete amino acid sequences were determined. Gigantoxins II (44 residues) and III (48 residues) have LD(50) (against crabs) of 70 and 120 microg/kg, respectively, and are analogous to the known type 1 and 2 sea anemone sodium channel toxins, respectively. On the other hand, gigantoxin I (48 residues) is potently paralytic to crabs (ED(50) 215 microg/kg), although its lethality is very weak (LD(50)>1000 microg/kg). Interestingly, gigantoxin I has 31-33% homologies with mammalian epidermal growth factors (EGFs), with the same location of six cysteine residues. In accordance with the sequence similarity, gigantoxin I exhibits EGF activity as evidenced by rounding of A431 cells and tyrosine phosphorylation of the EGF receptor in the cells, although much less potently than human EGF. Gigantoxin I is the first example of EGF-like toxins of natural origin. PMID- 12565743 TI - Intramuscular administration of antivenoms in experimental envenomation by Bothrops asper: comparison between Fab and IgG. AB - The efficacy of intramuscular (im) administration of sheep Fab and IgG antivenoms was assessed in a mouse experimental model of envenomation by Bothrops asper, in order to test if the more rapid absorption of Fab improves neutralization. Both antivenoms were adjusted to have a similar neutralizing potency in assays involving preincubation of venom and antivenom. Neither antivenom was effective in neutralizing lethality, nor in prolonging the time of death, in mice injected with either 3, 2 or 1.5 LD(50)s of venom by the intraperitoneal (ip) route, in experiments in which antivenoms were administered im immediately after envenomation. Antivenoms were effective in the neutralization of defibrinating activity, even if treatment was performed 30 min after envenomation, with no differences between IgG and Fab. Regarding neutralization of local effects, i.e. myonecrosis and hemorrhage, im administration of antivenoms at a site distant from the venom-injection site was completely ineffective in reducing the extent of local tissue damage. However, partial neutralization of these effects was achieved if antivenoms were administered im at the same site of venom injection, provided treatment was performed immediately after envenomation. Fab antivenom was slightly more effective than IgG antivenom in the neutralization of myotoxicity under these conditions, although a similar efficacy was observed between these antivenoms regarding neutralization of hemorrhagic effect. Our observations do not evidence major differences in the neutralizing ability of Fab and IgG antivenoms when applied by the im route, and do not support the hypothesis that im administration of Fab antivenoms constitutes an effective alternative to treat B. asper envenomations. PMID- 12565744 TI - Studies of the effect of Psi-APONIN from Nannochloris sp. on the Florida red tide organism Karenia brevis. AB - Studies were conducted on the conditions under which the red tide organism, Karenia brevis (a.k.a., Gymnodinium breve), was treated with Nannochloris sp. The latter organism is known to produce cytolytic agents called Apparent Oceanic Naturally Occurring Cytolin (APONINs). Conventional wisdom might suggest that brevetoxins would be released upon destruction of the single-celled dinoflagellate K. brevis and that efforts to treat red tide outbreaks would lead to release of brevetoxins and enhanced toxicity toward marine species. Earlier studies described conditions by which K. brevis cells were converted to a non motile form when cultures of K. brevis were treated with an isolate (Psi-APONIN) produced by Nannochloris sp. but when centrifuged only a small amount of the toxin was released. The present study confirms that the toxin is not released when the K. brevis is undisturbed, however, when the culture is stressed (stirred with a magnetic stirring bar for 24, 48, and 72h) toxin was released, and the toxicity could be measured using a Microtox analyzer. In the study, it was found that at as few as eighty cells of K. brevis produced a toxic effect of 20% as measured by the effect on Vibrio fischeri. Nannochloris sp. had no effect on the bacteria used in the Microtox analyzer, nor did interaction of Nannochloris sp. with K. brevis in the short term. This effect is presumed to be due to the production of Psi-APONIN and conversion of K. brevis to a non-motile or resting form. PMID- 12565745 TI - Epidemiology of snakebite in a central region of Brazil. AB - The aim of this article is to acquire knowledge about the aspects of snakebite epidemiology in a central region of Brazil. From 1993 to 1995, 90 cases of Crotalinae (Bothrops and Crotalus genera) and two cases of Micrurus snakebite were attended to in a general hospital. Epidemiological information about 73 out of the 90 Crotalinae victims was prospectively collected from interviews with the patients and/or their companions. Data from medical records were obtained for the 17 remaining cases. The snakes of Bothrops, Crotalus, and Micrurus genera were responsible for 74, 24 and 2% of the accidents, respectively. Most of the Crotalinae accidents occurred from October to March (68%) and from 06:00 to 12:00 a.m. (93%). Males (89%) and patients between 20 and 30 years-old (27%) were the most common victims. The main bite sites were: foot (24%), leg (23%), hand (22%) and ankle (21%). Among the 73 interviewed Crotalinae victims, farm workers were bitten more frequently (53%). The accidents often occurred during work (59%), and 90% of the patients wore footwear, but 30% were wearing only sandals. Tourniquet, squeezing, suction of the bite site and magic blessing were attempted in 47, 38, 8 and 10% of cases, respectively. PMID- 12565746 TI - Bibliography of toxinology. PMID- 12565747 TI - Protection against dermonecrotic and lethal activities of Loxosceles intermedia spider venom by immunization with a fused recombinant protein. AB - We report the use of a recombinant Loxosceles intermedia spider protein in the form of a fusion protein as an antigen for immunization in rabbits and mice. The aim is to produce model protective antisera in these animals against dermonecrotic and lethal activities of the venom from the Brazilian spider responsible for 3000 cases, reported annually, of spider bites in South Brazil. A protein homologous to the dermonecrotic toxin was cloned from a cDNA expression library made with L. intermedia venom glands, expressed in E. coli cells as a fusion protein with beta-galactosidase and the recombinant protein (Li-rec protein) was purified by molecular filtration and affinity chromatography [Kalapothakis et al., Toxicon (2002) in press]. The Li-rec protein was characterized and used as an antigen to generate antibodies in rabbits and mice. These specifically raised antibodies recognized the native venom. In vitro neutralization assay of lethal effects indicated that 1 ml of rabbit serum raised against Li-rec protein was able to neutralize 25 LD(50) of the whole venom. In vivo protection experiments, the fusion proteins induced a long-term protection in rabbits against the dermonecrotic activity of the native venom. Immunized mice were challenged with various doses of the Loxosceles venom. Mice were fully protected against 2.5 LD(50) of venom. This result provides basic data for the use of such recombinant spider proteins as immunogens in the development of anti venoms for clinical use or can be used as a vaccine providing efficient immune protection against L. intermedia venom. PMID- 12565748 TI - Isoform-specific effects of a novel BmK 11(2) peptide toxin on Na channels. AB - BmK 11(2) is a 7216Da polypeptide toxin purified from the venom of the scorpion Buthus martensii Karsch. Nanomolar concentrations of the toxin prolong amphibian nerve action potentials without attenuation of the amplitude. The pharmacological action of the toxin and its sequence similarity to other alpha-scorpion toxins suggest that BmK 11(2) selectively alters voltage-gated Na channels. In order to test whether BmK 11(2) preferentially modulates the gating or kinetics of certain channel isoforms, we applied BmK 11(2) to muscle, heart and neuronal Na channels. 100nM BmK 11(2) increased the peak current amplitude of skeletal muscle (micro1) and neuronal (N1E-115) Na currents by 40 and 20%, respectively, and reduced the cardiac Na (hH1) current by 15%. The toxin slowed current decay of all isoforms, most prominently in N1E-115 (tau(BmK)/tau(Control)=12), micro1 (11), and less so for hH1 (1.3). BmK 11(2) shifted the voltage dependence of activation of micro1 and N1E-115 currents. BmK 11(2) had no effect on steady-state inactivation, use dependent availability, and the kinetics of entry into slowly recovering inactivated states. PMID- 12565749 TI - Detection of five new hydroxyl analogues of azaspiracids in shellfish using multiple tandem mass spectrometry. AB - The polyether dinoflagellate toxins, azaspiracids, are responsible for azaspiracid poisoning (AZP), a new human toxic syndrome arising from the consumption of shellfish. To date, five azaspiracids have been isolated and fully structurally elucidated, including, AZA1, its 8-methyl and 22-demethyl analogues, AZA2 and AZA3, respectively, and two hydroxyl derivatives of AZA3, named AZA4 and AZA5. Using a recently developed method involving liquid chromatography with multiple tandem mass spectrometry (LC-MS(n)), five new azaspiracids, AZA7-AZA11, have been found in mussels (Mytilus edulis). AZA6 is a positional isomer of AZA1 and four of the new compounds are isomers with a mass of 857.5 amu. AZA7 and AZA8 are hydroxyl analogues of AZA1 while AZA9 and AZA10 are hydroxyl analogues of AZA6. AZA11 is a hydroxyl analogue of AZA2. The separation of all 11 azaspiracids was achieved using isocratic reversed phase liquid chromatography using a combination of eluent additives, trifluoroacetic acid and ammonium acetate. The ion-trap MS experiments, with electrospray ionisation, involved the fragmentation of the protonated molecule [M+H](+), trapping and fragmenting the product ions due to the loss of a water molecule [M+H-H(2)O](+), together with mass spectral data analysis that included the characteristic A-ring fragmentation for each compound. PMID- 12565750 TI - Snakebite-reappraisal of the situation in Eastern Nepal. AB - Four thousand and seventy eight cases of snakebite, occurring between January and December 2000, were analysed for clinical and epidemiological features. Cases of about 379 had features of envenoming and 81 died. All the victims with systemic envenoming had neurotoxicity. No case with coagulopathy was recorded. Snakebite was more frequent between the ages of 10 and 40 years (76%) and in males (73%). The majority (80%) of the snakebites were observed during the monsoon. Seventy percent of the bites with clinical features of envenoming occurred between 1400 and 2200 h. Five thousand eight hundred and fifty nine vials of polyvalent antisnake venom were used. Case fatality rate varied in the ten centres surveyed. It was as low as 3% in some to as high as 58% in others. Overall death rate among all snakebite cases was 2%. PMID- 12565751 TI - Occurrence of paralytic shellfish poison (PSP) in the starfish Asterina pectinifera collected from the Kure Bay, Hiroshima Prefecture, Japan. AB - Assays were made for paralytic toxicity of marine invertebrates inhabiting at the coasts of Hiroshima Bay, where the infestation of bivalves such as cultured oysters with paralytic shellfish poison (PSP) has been occurred. The starfish Asterina pectinifera collected at the estuary of Nikoh River, Hiroshima Bay, was found to contain moderate levels of paralytic toxicity. Its highest toxicities as PSP found on July 30, 1999 were 12.5 MU/g for whole body, 11.0 MU/g for integument tissues and 3.9 MU/g for viscera, respectively. The toxicity of integument was changed from 3.6 to 11.0 MU/g in 1 year. Its paralytic toxin principles were identified as PSP toxins, composing mainly from saxitoxin (STX) group toxins such as carbamoyl-N-hydroxy neosaxitoxin (hyneoSTX), and STX, by HPLC and LC-MS, accounting for over 90 mol%. The PSP toxins contained in the starfish A. pectinifera considered to be transferred from bivalves or detritus living in the same area, which were contaminated with PSP. However, the involved pathway may be different from that of Asterias amurensis which was infested directly through food chain from its food bivalves, for its toxin pattern. PMID- 12565752 TI - The first identification of the rare cyanobacterial toxin, homoanatoxin-a, in Ireland. AB - The first identification of the rare cyanobacterial neurotoxin, homoanatoxin-a, in Ireland is reported. A sensitive fluorimetric liquid chromatographic (LC) method was applied to the analysis of homoanatoxin-a in the low microg/l range. The analysis of the anatoxins in water samples required weak cation exchange solid phase extraction, fluorimetric derivatisation with 4-fluoro-7-nitro-2,1,3 benzoxadiazole (NBD-F), and isocratic reversed-phase LC. Confirmation of toxin identity was made using LC with electrospray mass spectrometry (MS) of the NBD derivatised homoanatoxin-a as well as LC-MS/MS of the free toxin. Application of the fluorimetric LC protocol to examine cyanotoxins in 20 Irish lakes resulted in the detection of homoanatoxin-a at four locations, Lough Sillan (24 microg/l), Inniscarra Reservoir (34 microg/l), Lough Key (12 microg/l), Caragh Lake (1.4 microg/l). An improved procedure for the isolation of homoanatoxin-a from cyanobacteria was also developed and confirmation of homoanatoxin-a was achieved by chromatographic and mass spectral comparison with authentic toxin isolated from a laboratory clone culture of Planktothrix (formerly Oscillatoria) formosa. PMID- 12565753 TI - Expression of a functional recombinant Phoneutria nigriventer toxin active on K+ channels. AB - PnTx3-1 is a peptide isolated from the venom of the spider Phoneutria nigriventer that specifically inhibits A-type K(+) currents (I(A)) in GH(3) cells. Here we used a bacterial expression system to produce an NH(2)-extended mutant of PnTx3-1 (ISEF-PnTx3-1) and tested whether the toxin is functional. The recombinant toxin was purified from bacterial extracts by a combination of affinity and ion exchange chromatography. The recombinant toxin blocked A-type K(+) currents in GH(3) cells in a fashion similar to that observed with the wild-type toxin purified from the spider venom. These results suggest that recombinant cDNA methods provide a novel source for the production of functional Phoneutria toxins. The recombinant ISEF-PnTx3-1 should be useful for further understanding of the role of A-type K(+) currents in biological processes. PMID- 12565754 TI - Cross reactivity of three antivenoms against North American snake venoms. AB - The antivenom in the United States today is in short supply, expensive and may not even be the most effective in neutralizing venoms from snakes in certain geographical locations. The ED(50) is considered to be the best indicator of antivenom efficacy, however, other tests are needed. In this study, three antivenoms (Antivipmyn (Fab(2)H), Crotalidae Polyvalent Immune Fab (Ovine) (FabO) and UCV (FabV) were used to test the effectiveness of neutralization of eight venoms (Agkistrodon piscivorus piscivorus, Bothrops asper, Crotalus adamanteus, C. durissus durissus, C. horridus atricaudatus, C. h. horridus, C. atrox, and C. molossus molossus). Four different assays were used to study the efficacy of the antivenoms: the antihemorrhagic, antigelatinase, antifibrinolytic and antihide powder azure. Fab(2)H antivenom was more effective in neutralizing the enzymatic activities of these eight venoms than FabO and FabV antivenoms. PMID- 12565755 TI - Separation of the components of type A botulinum neurotoxin complex by electrophoresis. AB - Clostridium botulinum neurotoxins (BoNTs) are the most toxic substances known. They exert potent neuroparalysis on vertebrates. C. botulinum produces seven serotypes of neurotoxin (A-G). BoNT/A, found in bacterial cultures of C. botulinum type A, is produced as a complex with a group of neurotoxin associated proteins (NAPs). Botulinum neurotoxin complex is the only known example of a protein complex where a group of proteins (NAPs) protect another protein (BoNT) against the acidity and proteases of the stomach. Here, we used sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for separation and identification of the constituent proteins of BoNT/A complex. A range of homogenous and gradient SDS-PAGE gels was used to resolve the BoNT/A complex. These gels were run under constant voltage and constant current conditions. The molecular weight and relative amount of each protein band were determined. On a 12.5% homogenous SDS-PAGE under reducing conditions, seven protein bands were identified with average molecular weights of 118, 106, 90, 56, 36, 23 and 17 kDa. The relative amounts of these seven proteins were determined densitometrically as 10, 6, 13, 27, 22, 13 and 8%, respectively. The separation and identification of BoNT/A complex will help in understanding the molecular structure and function of BoNT/A NAPs and their interaction with the toxin, in the toxico-infection process of the botulism diseased state. In particular, the stoichiometry of the individual components is established for a typical preparation of BoNT/A complex. Furthermore, the studies reported here identify the most favorable conditions for the baseline resolution of BoNT/A NAPs proteins for other workers in this field. PMID- 12565756 TI - Oxidative stress associated hepatic and renal toxicity induced by ricin in mice. AB - Ricin a glycoprotein from the Ricinus communis seeds, is known to have diverse toxic effects on cells of different visceral organs. We have studied the hepatotoxicity, nephrotoxicity, and oxidative stress following i.p. administration of ricin (25 microg/kg) in Swiss albino male mice. The results of this study revealed that activities of various enzymes like glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (gamma-GT), and lactate dehydrogenase (LDH) were increased in plasma, liver, and kidney tissues indicating damage in liver and kidney. Blood urea level was also increased. However, blood creatinine and bilirubin were not altered. Lipid peroxidation increased to 49 and 25% in hepatic and renal tissue. Total non protein sulfhydryl content decreased in plasma (12%), hepatic (29%), and renal (16%) tissues. Superoxide dismutase activity decreased significantly in liver (43%) and kidney (37%). The activity of glutatione peroxidase was also decreased. The decrease was more prominent in kidney than liver. A significant increase, 20 to 27% in the activity of catalase was observed in plasma, liver, and kidney. These results indicate that ricin produces hepatoxicity, nephrotoxicity, and oxidative damage at 24 h of post treatment. The hepatotoxicity was more prominent than nephrotoxicity. PMID- 12565757 TI - GTX(4) imposters: characterization of fluorescent compounds synthesized by Pseudomonas stutzeri SF/PS and Pseudomonas/Alteromonas PTB-1, symbionts of saxitoxin-producing Alexandrium spp. AB - Saxitoxins, the etiological agent of paralytic shellfish poisoning, are synthesized by dinoflagellates and cyanobacteria. Several reports indicate that bacteria are capable of saxitoxin synthesis. Two bacterial strains were isolated from saxitoxin-producing dinoflagellates, Alexandrium tamarense and A. lusitanicum (=Alexandrium minutum), and grown under a variety of culture conditions including those previously reported to induce saxitoxin synthesis in bacteria. Five fluorescent compounds were accumulated by the bacteria that had HPLC-FLD retention times similar to a reference standard of GTX(4), one of the saxitoxin congeners. However, we were unable to detect GTX(1), the epimeric partner of GTX(4), in the bacterial samples. The GTX(4) standard was hydrolyzed by NaOH/heat treatment but four of the bacterial compounds were stable. Unlike GTX(4), none of the five bacterial compounds were detectable by HPLC-FLD following electrochemical oxidation. The fluorescence emission spectrum of each of the five bacterial compounds was unique and readily discernable from the spectrum of GTX(4). None of the samples containing the putative GTX(4) toxin yielded positive results when analyzed by a 3H-saxitoxin receptor-binding assay for saxitoxin-like activity. We cannot rule out the possibility that these bacteria produce saxitoxins, however, our data clearly demonstrate that they accumulate at least five different fluorescent compounds that could be easily mistaken for GTX(4). We conclude that these five fluorescent compounds are GTX(4) imposters and that fluorescence scanning and chemical/heat stability should, at a minimum, be incorporated into HPLC-FLD protocols for identification of saxitoxins. PMID- 12565758 TI - Interaction between microcystins of different hydrophobicities and lipid monolayers. AB - Microcystins (MC) are a group of amphiphatic peptide hepatotoxins and protein phosphatase inhibitors produced by certain cyanobacteria (blue-green algae). Microcystins are believed to require an active transport mechanism to penetrate the plasma membranes of animal cells. In this study the surface barostat technique showed that two more hydrophobic microcystins MC-LF, containing Leu and Phe, and MC-LW, containing Leu and Trp, had a higher surface activity on an egg phosphatidylcholine-cholesterol (7:3, molar ratio) monolayer as compared to that of a more hydrophilic variant MC-LR, containing Leu and Arg. Fluorescence anisotropy measurements of 1-[4-(trimethylamine)phenyl]-hexa-1,3,5-trien (TMA DPH) were used to assess changes in the fluidity or lipid packing of model membranes in the presence of toxins. All three toxins caused a decrease in the steady-state anisotropy of TMA-DPH, suggesting that the toxins interacted with the membranes. The change in anisotropy was more pronounced for MC-LF and MC-LW than for MC-LR. Moreover, the fluorescence emission maximum of Trp in MC-LW was shifted slightly towards a shorter wavelength and the intensity was enhanced when allowed to interact with lipid vesicles, suggesting that the single Trp in MC-LW moved into a more unpolar environment when interacting with the vesicles. The differences between hydrophilic and hydrophobic microcystins could result in changes in organotropism, toxicokinetics and bioaccumulation. PMID- 12565759 TI - The efficacy of two antivenoms against the venom of North American snakes. AB - Mortality due to snake envenomation is not a major problem in the United States with approximately 8-12 deaths per year, but envenomation is a serious problem that can result in functional disability, loss of extremities, and a costly recovery. Physicians encounter different clinical situations with each new snakebite victim because of the geographical variations in snake venoms. The best and most acceptable form of treatment is the use of antivenom; however, it must be administered as soon as possible since it is not so effective at reducing local signs of envenomation such as necrosis. The antivenom in the United States is in short supply, expensive and may not even be the most effective for neutralizing all North American snake venoms. In this study, we tested two antivenoms. The first was a Crotalidae Polyvalent Fab fragment with Ovine origin (FabO) manufactured in London, and the second was Antivipmyn, a Mexican manufactured antivenom that is F(ab')(2) fragment produced in horse (Fab(2)H). The efficacy of the two antivenoms was tested with 15 different snake venoms found in North America. Three different assays were used to test the efficacy of the antivenoms, the in vivo serum protection test (ED(50)), antihemorrhagic and anticoagulant. The Fab(2)H antivenom was most effective in neutralizing the hemorrhagic activity of 78% of the hemorrhagic venoms used in this study. In the ED(50) assay, the Fab(2)H antivenom was effective in neutralizing all venoms used in this study, while FabO neutralized all but C. m. molossus venom. However, in most cases, FabO required less antivenom than Fab(2)H antivenom to neutralize three LD(50). PMID- 12565760 TI - Relationship between plasmatic levels of various cytokines, tumour necrosis factor, enzymes, glucose and venom concentration following Tityus scorpion sting. AB - A sandwich enzyme-linked immunosorbent assay was developed for measuring Tityus venom levels in plasma. The method proved capable of distinguishing patients with only local symptoms from controls, and was used to quantify venom in 205 accidental human envenomations. Our results show that the severity of envenoming is related to the patient plasma venom concentration. This depends on time elapsed between the sting and when the plasma was drawn. We observed that 46 and 49% of patients with moderate to severe symptoms (MS, n=41) showed hyperamylasemia and hyperglycemia, respectively. In addition, 39% of cases with MS symptoms had partial thromboplastin time values prolonged or shorted and 6.5% of patients with local symptoms (LS, n=164) had only prolonged prothrombin time values. Interleukin 6 (IL6) increased significantly in patients with MS symptoms. IL6 values increased with hyperamylasemia, envenoming severity and time hyperamylasemia. PMID- 12565761 TI - Renal effects of supernatant from rat peritoneal macrophages activated by microcystin-LR: role protein mediators. AB - We have demonstrated previously that microcystin-LR promoted some renal alterations using the isolated perfused rat kidney preparation. However, these effects were not proved to be direct or indirect. The aim of the current work is to examine the renal effects promoted by supernatants from rat macrophages stimulated with microcystin-LR and the role of inflammatory mediators. Peritoneal macrophages were collected previously and were incubated for 1h in fresh medium (control) and in medium containing microcystin-LR. Dexamethasone, quinacrine, thalidomide and cycloheximide were administered 30 min before microcystin-LR. Supernatants of macrophages stimulated with or without pharmacological inhibitors were added on the perfused rat kidney model. The infusion of macrophages supernatants stimulated by microcystin-LR caused significant increases in renal vascular resistance (C: 4.93+/-0.33 vs T: 5.15+/-0.21), glomerular filtration rate (C: 0.559+/-0.008 vs T: 0.978+/-0.15) and urinary flow (C: 0.16+/-0.01 vs T: 0.23+/-0.03). Cycloheximide, quinacrine and dexamethasone blocked these effects and thalidomide blocked renal vascular resistance. Macrophages stimulated by microcystin-LR release mediators capable of promoting nephotoxicity in isolated perfused rat kidney. Phospholipase A(2), TNF-alpha and other protein mediators appear to be involved on its renal toxic mechanism. PMID- 12565762 TI - Stimulation of GABA release by scorpion venom in an isolated synapse in the crayfish (Astacus leptodactylus). AB - Effects of various types of scorpion venom on gamma-aminobutyric acid (GABA) release were studied in an isolated synapse in the crayfish. Post-synaptic GABA induced currents were recorded to monitor the GABA release from the pre-synaptic site. In 20mM tetraethylammonium (TEA) chloride solution the GABA-induced currents increased 71%. Exposing the preparations to Leiurus quinquestriatus hebraeus, Leiurus quinquestriatus quinquestriatus or Tityus serrulatus venom (0.1mg/ml) increased GABA-induced currents 4-5 fold. The effect was present in the presence of tetrodotoxin (TTX) but diminished significantly when verapamil was applied. Exposing the preparations Androctonus australis or Buthus tamulus venom did not affect the GABA-induced currents. The results indicate that stimulation of the GABA release by some of the scorpion venoms may partly be due to a possible block of pre-synaptic potassium channels, but not due to an abnormal increase in sodium channel activation. PMID- 12565764 TI - Erectile function after radical prostatectomy: a review. AB - OBJECTIVES: Although the high rate of erectile dysfunction (ED) following prostatectomy is well recognised, the aetiology and pathophysiology have not yet been fully elucidated. We examined the current literature as to aetiology, treatment and possible prevention of ED following prostatectomy. METHOD: Review of the literature by a Medline search. CONCLUSION: The most important predictors of erectile function are pre-operative erectile function and the nerve sparing nature of the procedure. The former is determined by age and vascular risk factors whereas the latter is decided by the stage of the tumour and the skill of the surgeon. The value of intraoperative nerve mapping seems limited and the importance of nerve grafting is uncertain. Natural recovery of erection can take as long as 24 months. Patients complain about a lack of professional support. Symptomatic therapy may be applied according to the current general standards of treatment in men with ED. PMID- 12565765 TI - What robotics in urology? A current point of view. PMID- 12565766 TI - Current status of robotics in urologic laparoscopy. AB - Urology has continuously embraced novel technologies like laparoscopy that reduce patient morbidity yet maintain an excellent standard of care. Because of limitations on maneuverability, operative vision, manual dexterity, and tactile sense, laparoscopy can be more difficult to perform than corresponding tasks in open surgery. To potentially increase clinical applicability of laparoscopy, robots that enhance operative performance have recently been introduced for a variety of laparoscopic procedures such as laparoscopic radical prostatectomy, pyeloplasty, and even laparoscopic cystectomy and neobladder construction. While these robots have generated excitement and many robotic applications have been described, the benefit of the advanced technology in expanded series of patients remains largely unknown. In addition, the ability of telerobotics to be used by surgeons inexperienced in conventional laparoscopy is also poorly understood. This review compares current features of available robots, advantages and limitations of robots, the emerging clinical applications, and the future potential of robotics in urology. PMID- 12565767 TI - Intraoperative and perioperative morbidity of contemporary radical retropubic prostatectomy in a consecutive series of 1243 patients: results of a single center between 1999 and 2002. AB - OBJECTIVES: To up date counselling of patients in an experienced center, we assessed intraoperative and perioperative complications in a consecutive series of contemporary radical retropubic prostatectomy for localized prostate cancer. METHODS: In a prospective study, we analyzed all intraoperative and perioperative complications within 30 days in a consecutive series of 1243 patients undergoing radical prostatectomy between January 1999 and February 2002. All adverse events were graduated in major and minor complications by their severity and sequel. RESULTS: There were no deaths. Overall, 996 patients (80.2%) were not affected by any complication. Major complications were observed in 50 patients (4.0%), minor complications in 197 (15.8%). Pelvic lymphadenectomy was performed in 861 (69.3%) patients. This procedure was associated with a significantly higher rate of lymphoceles requiring a drainage, 4.2% versus 0.3% (p<0.006) and a higher rate of deep venous thrombosis, 1.4% versus 0.5% (p<0.2), respectively. CONCLUSION: Radical retropubic prostatectomy is a safe surgical procedure. Postoperatively the majority of our patients was not compromised by any complication within 30 days. Furthermore, due to a higher rate of lymphoceles and thromboembolic events the indication for pelvic lymphadenectomy should be considered carefully. PMID- 12565768 TI - A longer duration of neo-adjuvant combined androgen blockade prior to radical prostatectomy may lead to lower tumour volume of localised prostate cancer. AB - OBJECTIVES: To analyse tumour volume (TV) in clinically localised prostate cancer patients treated with neo-adjuvant combined androgen blockade (CAB) therapy prior to radical prostatectomy. PATIENTS AND METHODS: Two hundred consecutive patients treated between 1996 and 2000 were retrospectively analysed. Fifty patients underwent radical prostatectomy alone and 45 were treated with CAB for 1-3 months, 83 for 4-6 months and 22 for more than 6 months before surgery. Logistic regression analysis was performed to identify the strongest independent prognosticator of organ-confined disease. RESULTS: No evidence of residual cancer was found in 11 specimens (5.6%). Regarding TV, 20 specimens showed less than 0.1cc, 33 between 0.1 and 0.49cc and 86 more than 0.5cc. Smaller TV was found in CAB-treated patients. Significant correlation was observed between treatment duration and TV. In logistic regression analysis, only CAB duration and TV were significantly correlated with organ-confined disease. CONCLUSIONS: Prominent regressive features and lower TV were found after neo-adjuvant CAB. It seems that more prolonged treatment may lead to greater tumoural regression. Only tumour burden and length of CAB therapy were independent variables significantly correlated with pathologically localised prostate cancer. PMID- 12565769 TI - The value of dynamic contrast enhanced power Doppler ultrasound imaging in the localization of prostate cancer. AB - OBJECTIVES: The objective of this study is to define enhancement characteristics that correlate to the presence of prostate cancer (PCa) and to evaluate the value of these characteristics in the localization of prostate cancer. METHODS: 29 patients with proven prostate malignancy, scheduled for radical prostatectomy, underwent an ultrasound examination prior to surgery. A bolus injection of contrast agent was administered intravenously. The distribution of the contrast enhanced blood to the prostate was monitored using Transrectal Contrast Enhanced Power Doppler Ultrasound. Fixed protocols and settings were used for all patients. The percentage of a selected area that showed enhancement was observed in time. The resulting enhancement curves were described using the parameters time to start, time to the maximum of the enhancement, the maximum value of the enhancement, and the rise time of the enhancement. Three evaluation-protocols divided the prostate into a number of areas of interest: into two areas using the Left-Right (LR) and Dorsal-Ventral (DV) protocols and into four areas using the Quadrant-protocol (Q). The enhancement parameters of the areas of interest were compared to identify the most affected area. The results were compared to the histopathological findings. RESULTS: For the LR-protocol, the minimal time to peak proved to be the most predictive parameter for selecting the major malignant area. 78% of the patients were diagnosed correctly (N=23). Accurate localization of the major malignancy in either the ventral or dorsal side of the prostate was not feasible using the current protocol. CONCLUSIONS: Malignancies can be accurately localized in either the left or the right side of the prostate based on the time to the maximum of the enhancement. An accurate discrimination between malignancies in either the dorsal or ventral side of the prostate cannot be performed. This is most likely due to anatomical differences between the dorsal and ventral area. PMID- 12565770 TI - The influence of serial sections, immunohistochemistry, and extension of pelvic lymph node dissection on the lymph node status in clinically localized prostate cancer. AB - OBJECTIVES: Pelvic lymph node metastases indicate a poor prognosis for patients with clinically localized prostate cancer. The aim of the study was to investigate the value of extended histopathological techniques considering the extent of pelvic lymphadenectomy and preoperative risk factors. METHODS: Total of 194 patients with prostate cancer were examined. At first all patients had a sampling of the sentinel lymph nodes (SLN) followed in most cases by a modified or extended pelvic lymphadenectomy. Step sections, serial sections and immunohistochemistry (IHC, pancytokeratin antibody) were analyzed in all SLN and so-called non-SLN of the first 100 patients. Later serial sections and IHC of non SLN were left out. RESULTS: In 26.8% lymphatic metastases were found. The detection rate of lymph node-positive patients depend significantly on the chosen extension of pelvic lymphadenectomy. Limiting the histopathological investigation to the lymph node specimen of the obturator fossa only 44.2% of lymph node positive cases would have been identified. An additional inclusion of all lymph nodes surrounding the external iliac vessels improves the sensitivity to 65.4% (46.7% and 73.3% for the first 100 patients). Compared to the extension of pelvic lymphadenectomy the diagnostic gain of serial section and IHC (13.8% versus 53.3%) was comparably low. CONCLUSIONS: The extension of pelvic lymph node dissection is of outstanding value for the identification of node-positive patients. Limiting the number of lymph nodes to the ones with the highest probability of bearing lymphatic spread (SLN) makes the use of extensive histopathological techniques more feasible. PMID- 12565771 TI - Prognostic value of MCM2 immunoreactivity in stage T1 transitional cell carcinoma of the bladder. AB - OBJECTIVE: Due to the heterogeneous biologic behavior of stage T1 bladder carcinomas, there is a need for new markers allowing to assess the prognosis more accurately. To our knowledge, there are no reports on studies investigating minichromosome maintenance protein 2 (MCM2) expression in bladder carcinomas. Thus, we investigated the prognostic value of MCM2 immunoreactivity in stage T1 bladder tumors. METHODS: Fifty-four tumors were analyzed using Biochip microarrays. Also p53 and Ki67 antigen expression were examined. Immunohistochemical scores were compared with the clinical outcome. RESULTS: During a median follow-up of 43 months, tumor recurrence was registered in 43 and progression to stage T2 in 19 patients. Kaplan-Meier curves demonstrated that high-level MCM2 expression was significantly associated with early tumor recurrence when using a cutoff of 60% (p=0.0035 by log-rank test), and with early tumor progression when using a cutoff of 20% (p=0.0454). There was no relationship (p=0.604) between MCM2 and p53, but a tendentious relationship (p=0.082) between MCM2 and Ki67 antigen expression. MCM2 (p=0.006), Ki67 antigen (p=0.035) and p53 expression (p=0.049) as well as tumor grade (p=0.026) and age (p=0.025) were found significantly associated with recurrence-free survival by univariate Cox regression analysis, among which only Ki67 antigen expression (p=0.015) and age (p=0.019) proved to be of independent predictive value by multivariate analysis. Concerning tumor progression, MCM2 expression was identified as the only predictive parameter by log-rank test, but it was not of independent predictive value by multivariate analysis (p=0.101). CONCLUSION: Our data suggest that MCM2 expression may bear some prognostic relevance in stage T1 bladder carcinomas. PMID- 12565772 TI - Are false-positive urine markers for the detection of bladder carcinoma really wrong or do they predict tumor recurrence? AB - INTRODUCTION AND OBJECTIVES: A problem in the interpretation of noninvasive urine tests for detection of bladder carcinoma is the finding of false-positive results. Several authors have described that patients with false-positive results are at high risk for tumor recurrence or progression. Only few data are available for comparing the clinical course of patients with false-positive test results and patients with true-negative results. We studied whether patients with false positive results of various urine test had a higher recurrence rate than patients with true-negative results. METHODS: Urine samples from 61 patients without evidence of active bladder carcinoma were included. Of the 61 patients, 51 had a history of bladder cancer, and 10 underwent transurethral resection for suspect of bladder carcinoma but had negative pathologic findings. Immunocytology (Lewis X and 486p3/12) was performed on bladder washings, and BTAstat and NMP22 were performed on urine samples. RESULTS: During the follow-up period, 22 patients had one or more false-positive BTAstat test results, 25 patients had one or more false-positive NMP22 tests, 42 patients had at least one false-positive Lewis X test, and 11 patients had one or more false-positive 486p3/12 test. During a follow-up period of 3-39 months (median, 17.6 months) four patients expected a tumor recurrence. Among patients with false-positive urine test results 2 of 22 (9.1%, BTAstat), 2 of 25 (8%, NMP22), 4 of 42 (9.5%, Lewis X), and 3 of 11 (27.2%, 486p3/12) suffered from tumor recurrence. In contrast, among patients with true-negative test results 2 of 39 (5.2%, BTAstat), 2 of 36 (5.6%, NMP22), 0 of 18 (0%, Lewis X), 1 of 50 (2.0%, 486p3/12) had a tumor recurrence. CONCLUSIONS: Patients with a false-positive urine test result do not generally have a greater risk of tumor recurrence or progression than patients with a true negative result. In our series, only patients with false-positive 486p3/12 test result had a higher recurrence rate. Our findings do not justify a more aggressive adjuvant treatment or surveillance for patients with false-positive urine tests. PMID- 12565773 TI - Polypropilene sling of the bulbar urethra for post-radical prostatectomy incontinence. AB - OBJECTIVE: We present details of a modification of the Stamey procedure using a polypropilene mesh to compress the corpus spongiosum of the bulbar urethra in the treatment of sphincteric incompetence following radical prostatectomy. MATERIALS AND METHODS: Between September 1999 and June 2000 nine patients 66-80 years old (mean age 74) with severe incontinence due to radical prostatectomy underwent the bulbourethral sling procedure with polypropilene (Prolene) mesh implant. After transperineal incision a 5cmx4cm rectangular prolene mesh was placed against the bulbar urethra and suspended by four prolene sutures transferred to a suprapubic incision through ligature carriers as in the four corner bladder and bladder neck suspension. If leakage recurred in the follow-up, a retightening procedure was performed in local anesthesia rescuing the prolene threads over a polypropilene mesh placed against the rectus fascia. Mean follow-up was 14 months (range 12 20). Continence status and post-void residual volume were evaluated after 1, 3 and 6 months post-operatively and successively every 3 months. RESULTS: At the present follow-up considering the results of the retightening procedure five out of nine patients were cured, two out of nine improved and two were unchanged. Five out of nine patients reported slight to mild perineal/genital discomfort which disappeared in four at three months post-operatively. No patients required mesh removal for complications. CONCLUSIONS: The male bulbourethral sling procedure using a polypropilene mesh is safe but further experience is needed to establish this procedure as plausible alternative for the treatment of post radical prostatectomy urinary incontinence. PMID- 12565774 TI - Percutaneous tibial nerve stimulation as neuromodulative treatment of chronic pelvic pain. AB - PURPOSE: Neuromodulative therapies have been used with moderate success in patients with chronic pelvic pain. Intermittent Percutaneous Tibial Nerve Stimulation (PTNS) is a new, minimally invasive treatment option, which has shown to significantly decrease accompanying pain complaints in patients with lower urinary tract dysfunction, such as urge incontinence or urgency/frequency. In our study, we evaluate the objective results of PTNS in patients with chronic pelvic pain as their main complaint. MATERIALS AND METHODS: In a prospective multicentre trial PTNS was evaluated in 33 patients with chronic pelvic pain. Effects were recorded by Visual Analogue Scale (VAS) for pain diaries, the McGill pain questionnaire and the SF-36 general quality of life questionnaire at baseline and after 12 weeks of treatment. Subjective (patients' request to continue chronic treatment to keep the obtained success) and objective responses (decrease in mean VAS >50% and VAS <3 after treatment) were evaluated. RESULTS: A subjective response was seen in 42% of all patients. In seven patients (21%) mean VAS decreased >50%, in six cases (18%) the decrease was >25%. After 12 weeks of treatment, seven patients (21%) ended up with a mean VAS <3. In all patients quality of life (SF-36) significantly improved, as did the total pain rate intensity (McGill). CONCLUSIONS: Despite very modest overall success rates and the need for placebo-controlled studies, PTNS may have a place in the treatment of patients with chronic pelvic pain who have already tried many other therapies and are left with no further option. PMID- 12565775 TI - Distribution of inflammation, pre-malignant lesions, incidental carcinoma in histologically confirmed benign prostatic hyperplasia: a retrospective analysis. AB - OBJECTIVES: We analyze our experience on BPH through 20 years of histopathological examinations performed by the same pathologist. METHODS: We retrospectively reviewed all histopathological examinations performed from January 1979 to December 1998 in patients undergoing surgery in our urological clinic who were diagnosed with BPH. We limited our evaluation to the following variables in each BPH case analyzed: inflammatory aspects associated with BPH, presence of focal acinar atrophy, atypical adenomatous hyperplasia (AAH), prostatic intraepithelial neoplasia (PIN), incidental prostate carcinoma (IC). These histological variables were analyzed according to some clinical parameters such as age, prostate volume and serum PSA. RESULTS: The study population was comprised of 3942 cases with histological diagnosis of BPH. The mean patient age was 68.85+/-7.67 years. In particular, inflammatory aspects were associated with BPH in a high percentage of cases (43.1% =1700 cases), predominantly as chronic inflammation. Observation of focal acinar atrophy significantly increased according to patient decade of age (p=0.027). There was a significant trend to increase with age decades (p=0.036) for high grade PIN. A significant difference was found in IC (T1a, T1b) distribution in the different decades of age and especially in regards to both T1a and T1b tumors, there was a trend to increase with patient age (p=0.020 and p=0.025, respectively). On the contrary, the distribution of inflammatory aspects (p<0.001) and AAH (p=0.003) significantly varied according to prostate volume, and particularly in regards to chronic inflammation, there was a trend to increase depending on the prostate volume (p=0.002). Only the presence of T1b tumor but not of the other histological parameters associated to BPH, was able to significantly influence serum PSA. CONCLUSION: In our analysis different histological variables associated to BPH are differently influenced by the age of patients and prostate volume, and they differently influence serum PSA levels. PMID- 12565776 TI - Transurethral microwave thermotherapy (TUMT) with the Targis System: a single centre study on 78 patients with acute urinary retention and poor general health. AB - OBJECTIVES: To evaluate the Targis System in men presenting with acute urinary retention, high prostate volume and high operative risk. MATERIALS AND METHODS: Between August 1997 and March 2001, a total of 78 patients in poor general health status presenting with large prostate glands and acute urinary retention secondary to BPH were treated with the Targis TUMT device. Mean age, mean prostate volume, and the percentage of patients who were able to urinate spontaneously after the procedure as well as mean peak and average flow rates and mean residual urine volume were evaluated. RESULTS: 68 (87.1%) of the 78 patients were able to urinate spontaneously three months after the procedure. In 5 (7.3%) of the 68 patients urinary retention recurred within two years. Following treatment, the mean peak flow rate in the 68 successfully treated patients was 11.1 ml/s, while the mean postvoid residual volume was 46 ml. CONCLUSION: Based on these data we recommend transurethral thermotherapy using the Targis System for patients in poor general health presenting with urinary retention and prostate volumes of more than 35 cc in whom TURP is not possible. PMID- 12565777 TI - Quantification of prostate shrinkage after microwave thermotherapy: a comparison of calculated cell-kill versus 3D transrectal ultrasound planimetry. AB - PURPOSE: To compare prostate shrinkage after transurethral microwave thermotherapy (TUMT) with calculated cell-kill. MATERIALS AND METHODS: The calculated cell-kill from 33 males with benign prostatic hyperplasia (BPH) treated with TUMT according to the ProstaLund Feedback Treatment (PLFT) method was compared to the post-treatment prostate volume change. The prostate volume was estimated with three-dimensional transrectal ultrasound (3D-TRUS) planimetry at baseline, 3, 6, and 12 months follow-up. A paired t-test was used to test the statistical significance of differences between the cell-kill volume and the prostate volume change. Linear regression was used to infer a relationship between the cell-kill and the 3D-TRUS data. The reproducibility of the 3D-TRUS method was assessed in repeated measurements. RESULTS: The mean prostate volume at baseline (N=33) was 56.1cm(3). After 3 (N=25), 6 (N=29) and 12 months (N=23), it was 45.5 cm(3), 39.7 cm(3), and 45.1cm(3), respectively. The corresponding average cell-kill volume was 16.4 cm(3), 17.1cm(3), and 17.2 cm(3), respectively. Predicted cell-kill volume was significantly larger than prostate shrinkage at 3 (p<0.0001), 6 (p=0.0002), and 12 months (p<0.0001), and showed a strong correlation at 3 and 6 months (r=0.74, p<0.0001). Correlation at 12 months was moderate (r=0.57, p=0.0041). Examination and investigation variability both averaged 2.5%. CONCLUSIONS: Cell-kill calculations of the PLFT method are proportional to the 3D-TRUS prostate shrinkage by a factor of 0.5 and have a precision of approximately +/-10 cm(3) for 90% of the patients during the first year after treatment. PMID- 12565778 TI - The impact of radiological anatomy in clearance of lower calyceal stones after shock wave lithotripsy in paediatric patients. AB - OBJECTIVE: To determine the factors affecting stone clearance after extracorporeal shock wave lithotripsy (ESWL) in children with lower calyceal stones. MATERIALS AND METHODS: Lower pole stone clearance was reviewed in 34 patients aged between 2 and 16 years (23 boys and 11 girls) treated with ESWL between 1989 and 2001 in our clinic. Renal anatomy was determined on standard intravenous urograms. The lower infundibulopelvic angle (IPA) was measured by two different methods based either on measuring the angle between vertical pelvis axis and vertical axis of lower infundibulum or finding the angle between the ureteropelvic axis and vertical axis of lower infundibulum. RESULTS: The mean age of the patients was 12.1+/-4.16 (2-16) years. The mean stone burden was found as 96.93+/-87.13 mm(2) and the mean shock wave number per session and power as 2631.4+/-593.1 and 17.57+/-1.1 kV, respectively. The stone-free rate was 55.9%. Fifteen (44.1%) cases had residual fragments retained in lower calices after lithotripsy and are still followed. The stone clearance was found to be unrelated to stone burden and infundibulum width (p=0.44 and p=0.34, respectively). However, a significant difference was present between mean lower pole infundibular length (p=0.0032) and lower IPA measurements according to both methods between stone-free cases and others. The most remarkable factor that had a significant influence on stone clearance was an acutely oriented infundibulum defined according to IPA-ureteropelvic axis angle determination method (p=0.00001) followed by Sampaio's pelvic axis method (p=0.0001). Only 1 (5%) patient was stone-free under 40 degrees and 1 (6%) case with an angle of 45 degrees had residual fragments in the former method. Similarly none of the cases had residual fragments over 90 degrees for Sampaio's method and 90% of the stone free cases had a more obtuse angle. Thus, we determined that a cut-off point of 40 degrees for the IPA-ureteropelvic axis method and 90 degrees for Sampaio's method was most useful to determine the clearance of lower pole fragments. CONCLUSION: The factors affecting stone clearance in paediatric patients are similar to adults. PMID- 12565779 TI - Lignocaine gel: does it cause urethral pain rather than prevent it? AB - OBJECTIVES: To test the hypothesis that the chemical content of lignocaine gel is the cause of urethral pain during its instillation into the urethra. METHODS: A prospective, double blind study was designed to test whether plain aqueous gel caused less delivery discomfort in the male urethra than 2% lignocaine hydrochloride gel (Instillagel). 100 consecutive, consenting male patients attending for flexible cystoscopy were recruited and randomised to receive 11 ml of 2% lignocaine hydrochloride gel or 11 ml of plain aqueous gel. Upon receiving the gel into the urethra, the patient was asked to score any associated discomfort by using a horizontal, 100mm, non-graphical, visual analogue scale. Statistical comparison between the groups was made using the non-parametric Mann Whitney U-test. RESULTS: Statistical analysis by Mann-Whitney U-test showed a significant reduction in urethral delivery discomfort in those patients receiving plain gel compared to those receiving 2% lignocaine hydrochloride gel (p=0.039). CONCLUSIONS: This current study has shown that plain aqueous gel causes significantly less delivery discomfort in the male urethra than 2% lignocaine hydrochloride gel (Instillagel). PMID- 12565780 TI - Osteocalcin gene HindIII C/T polymorphism is a biomarker for prostate cancer and responsiveness to hormone therapy. AB - OBJECTIVES: Osteocalcin is a vitamin-K dependent protein which is related to the metabolism of bone and calcium. The formation or progression of prostate cancer is presumed to be associated with the osteocalcin gene. The most frequently seen polymorphism is HindIII which is located at the promoter region. HindIII is therefore a possible genetic marker in the search for the association between prostate cancer and normal control subjects. METHODS: In our study, a normal control group of 132 healthy people and 96 patients with prostate cancer were examined. The polymorphism was seen following polymerase chain reaction (PCR) based restriction analysis. RESULTS: The result revealed significant differences between normal individuals and cancer patients (p=0.034) and the distribution of the "CC" homozygote in the control group was higher than that in the patient group. No statistical differences were found in clinical staging and grading. The 54 patients who received hormone therapy were further categorized into response and non-response groups, statistical differences between these two groups were revealed (p=0.007, Fisher's exact test). CONCLUSIONS: Based on our results, we conclude that the HindIII polymorphism of the osteocalcin gene is a suitable genetic marker of prostate cancer which can be used in the prediction of the outcome of patients who receive hormone therapy. PMID- 12565781 TI - Pan-caliceal ileoneocystostomy-a variant of ileal ureter. PMID- 12565783 TI - Re: Knutson T, Pettersson S, Dahlstrand C. The use of biodegradable PGA stents to judge the risk of post-TURP incontinence in patients with combined bladder outlet obstruction and overactive bladder. Eur Urol 2002;42:262-7. PMID- 12565785 TI - Validity of questionnaires. PMID- 12565786 TI - Anaesthesia for tension-free vaginal tape. PMID- 12565790 TI - Adhesion molecules as susceptibility factors in spontaneous autoimmune thyroiditis in the NOD-H2h4 mouse. AB - Mononuclear cell infiltration of the thyroid is a prominent feature of chronic lymphocytic thyroiditis. Adhesion molecules play a major role in determining the localization of inflammatory mononuclear cells in the thyroid. Previous reports from animal models and human studies have described the thyroidal expression of adhesion molecules only late in clinical disease. In this study, we examined the distribution and kinetics of expression of E-selectin, VCAM-1, LFA-1, and ICAM-1 in the NOD-H2h4 mouse, a model of spontaneous autoimmune thyroiditis accelerated by dietary iodine. Mice were fed 0.015% NaI in their drinking water for 2, 4, 6, 8, and 16 weeks, and thyroids were removed, serially sectioned, and stained in an avidin-biotin-peroxidase assay. We found a dramatic increase in E-selectin and VCAM-1 expression on intrathyroidal endothelial cells after 16 weeks of iodine treatment. In addition, we describe for the first time that thyrocytes from the NOD-H2h4 mouse, and the parental NOD, constitutively express ICAM-1 independent of iodine treatment and prior to mononuclear cell infiltration of the thyroid gland. ICAM-1 was not detected on the thyrocytes of other untreated strains of mice, implicating expression of this adhesion molecule as a critical event in the recruitment of inflammatory mononuclear cells to the thyroid. PMID- 12565791 TI - Regulatory role of IL-10 in experimental obliterative bronchiolitis in rats. AB - Obliterative bronchiolitis (OB) affects over half of all chronic human survivors following lung or heart-lung transplantation. Respiratory epithelial cell injury, peribronchial inflammation, and proliferation of fibrovascular connective tissue causing airway occlusion characterize this lesion. Using a rat model of experimental OB, tracheas and mainstem bronchi from Brown-Norway or Lewis (LEW) rats were transplanted subcutaneously into LEW recipients. At 7 days, airway lumens of allografts showed minimal luminal obstruction but significant respiratory epithelial loss. By 14 days, allografts demonstrated marked peribronchial inflammation, nearly complete loss of respiratory epithelium, and extensive intraluminal proliferation of fibrovascular connective tissue, with a mean 58% reduction in airway cross-sectional diameter. However, isografts showed only limited peribronchial inflammation and no loss of airway lumen. When recipients of allotransplants were treated with anti-IL-10, OB developed more rapidly. As early as 7 days, there was marked histologic evidence of OB and a 43% reduction in mean cross-sectional area. Allograft animals that received 5 microg/day of recombinant IL-10 as a constant infusion on day 14 showed almost complete preservation of respiratory epithelium and only mild peribronchial inflammation with only a 15% reduction in airway cross-sectional area. These findings suggest that endogenous IL-10 plays a regulatory role in the development of experimental OB. PMID- 12565792 TI - Comparative study of adhesion molecule expression in cultured human macro- and microvascular endothelial cells. AB - Culture systems as models for disease are only valid as long as they are comparable to in vivo conditions. The phenotype of cultured endothelial cells (ECs) has only been sporadically compared to the corresponding phenotype in vivo. Thus, we compared by immunolocalization the endothelial expression of ICAM-1, VCAM, and E-selectin in vivo in stimulated/unstimulated human umbilical vein endothelial cells (HUVEC) as a model for macrovascular ECs and stimulated/unstimulated HPMEC (human pulmonary microvessel endothelial cells) as a model for pulmonary microvascular ECs with that in human lungs in vivo (normal and ARDS). Proinflammatory stimuli in vitro were used to stimulate conditions relevant for ARDS. ICAM-1 expression in stimulated HUVEC/HPMEC correlated well with in vivo expression (macro- and microvessels). For E-selectin, the staining pattern in macro/microvessels correlated moderately with unstimulated and well with stimulated HUVEC/HPMEC. For VCAM a good correlation was found for stimulated/unstimulated HUVEC and unstimulated HPMEC. The expression patterns in stimulated HUVEC corresponded well for all three molecules with those in vivo. Thus, the expression patterns in vitro are only partially transferable to in vivo conditions. The study suggests that E-selectin- and VCAM-coated beads could potentially serve in the isolation process of arteriolar and venular ECs. PMID- 12565793 TI - Differential distribution of fibroblast growth factor (FGF)-7 and FGF-10 in L arginine-induced acute pancreatitis. AB - The regenerative process of the pancreas after acute pancreatitis (AP) is characterized by acinar and ductal cell proliferation with synthesis and transient deposition of extracellular matrices. Various growth factors were reported to be highly expressed in AP, but their regulation has not yet been clarified. Fibroblast growth factor (FGF)-7, also known as keratinocyte growth factor (KGF), and FGF-10 are members of the FGF family and show high structural homology and similar biological characteristics. Both are mainly synthesized by mesenchymal cells and stimulate epithelial cells via KGF receptor (KGFR) which is a splice variant of FGFR-2. In the present study, we attempted to immunohistochemically determine the localization of FGF-7 and FGF-10 in pancreatic tissues of an L-arginine-induced rat pancreatitis model. Furthermore, highly specific KGFR antibodies were prepared and used for Western blot analysis and immunohistochemistry. In the normal pancreas, FGF-7 was localized in alpha cells of islets, but FGF-10 was not detected. KGFR was also localized in islet cells, ductal cells, and centroacinar cells in the normal pancreas. In the pancreatic tissues of rats with L-arginine-induced pancreatitis, FGF-7 was localized in alpha cells, whereas FGF-10 was expressed in vascular smooth muscle cells (VSMCs). KGFR was not expressed in centroacinar cells and its level decreased after L-arginine treatment. However, KGFR was detected instead in some acinar cells and VSMCs in addition to islet cells. These findings suggest that FGF-7 and FGF-10 contribute to the regeneration and differentiation of acinar cells and angiogenesis in AP through KGFR. PMID- 12565794 TI - Extracellular matrix oxidation modulates survival, NF-kappaB translocation, and MAPK activity in mesangial cells. AB - Inflammatory injury to the kidney may oxidize extracellular matrix proteins. These changes appear to impair adhesion of glomerular mesangial cells and might impact signaling mechanisms in these cells. In this study we evaluated how extracellular matrix oxidation could modulate the development of apoptosis as well as the activity of the transcription factor NF-kappaB and that of the MAPK family members ERK-1,2 and JNK. Exposure of mesangial cells to oxidized matrix enhanced the percentage of apoptotic cells. Western blot analysis revealed diminished levels of the phosphorylated form of ERK-1,2 in cells on oxidized matrix, while levels of phosphorylated JNK were increased. Cells on unmodified matrix were found to have rapid translocation of the p65 subunit of NF-kappaB, which was attenuated in cells on oxidized matrix. These findings suggest that extracellular matrix oxidation may impair survival of mesangial cells in association with decreased levels of pERK-1,2, increased levels of pJNK, and diminished nuclear translocation of the transcription factor NF-kappaB. PMID- 12565795 TI - MDM2 and CDK4 expression in carcinosarcoma of the esophagus: comparison with squamous cell carcinoma and review of the literature. AB - Certain tumors of the esophagus that display both sarcomatous and carcinomatous features have long been recognized. The nomenclature, classification, and histogenesis remain controversial and the microscopic differential diagnosis from other esophageal malignancies can be challenging, particularly in small biopsies. In this paper, we review the literature of carcinosarcoma and present two cases of esophageal carcinosarcoma, describing their salient histologic, immunohistochemical, and ultrastructural features. Also, we assess the expression of MDM2 and CDK4 in the carcinomatous and sarcomatous compartments of our cases and we compare them with the expression of these oncogenes in selected cases of esophageal squamous cell carcinoma with prominent stromal reaction. In both of our cases, identification of some epithelial ultrastructural and immunohistochemical features in cells of otherwise sarcomatous phenotype lends support to the common epithelial origin of these neoplasms. Moreover, positive staining for MDM2 and CDK4 in our cases with equally strong reactions in both carcinomatous and sarcomatous elements provides evidence of a role for these molecules in the pathogenesis of carcinosarcoma. In contrast, in cases of squamous cell carcinoma with prominent stromal reaction only the epithelial cells stained strongly for MDM2 and CDK4. These differences in the MDM2 and CDK4 immunohistochemical profile between carcinosarcomas and carcinomas of the esophagus may assist in their differential diagnosis. PMID- 12565796 TI - Hepatic expression of polymerase beta, Ref-1, PCNA, and Bax in WY 14,643-exposed rats and hamsters. AB - The hepatic levels of three protein markers of oxidative stress, polymerase beta, Ref-1, and PCNA, and of the pro-apoptotic protein, Bax, were quantitated after exposure to WY 14,643 (500 ppm in the feed) for 6 or 34 days in a rodent that is susceptible peroxisome proliferator (PP)-induced liver tumors (the Sprague Dawley rat) and in a rodent that is relatively resistant PP-induced liver tumors (the Syrian hamster). The analysis of detergent-extracted whole liver homogenates by immunoblotting showed a marked increase in the abundance of a 45-kDa variant of polymerase beta immunoreactivity and significant increases in the expression of Ref-1 and PCNA in WY 14,643-exposed rats. In contrast. WY 14,643-exposed hamsters expressed only trace levels of the polymerase beta variant and showed significant decreases in the expression of Ref-1 and PCNA. Long-term WY 14,643 exposure was associated with marked decreases in Bax expression in both species. Dose-response studies in the rat showed that the hepatic expression of the polymerase beta and Ref-1 were significantly increased after 6 days of exposure to WY 14,643 at levels of 5 and 50 ppm, respectively. The analysis of subcellular fractions of rat liver showed that the pathological increases in the levels of polymerase beta, Ref-1, and PCNA were especially prominent in mitochondria-enriched particulate liver subfractions. These results indicate that WY 14,643 exposure is associated with an increase in oxidative stress to the liver and that liver mitochondria are a major target of WY 14,643-associated liver damage. Our data are consistent with the hypothesis that the chronic overexpression of mutagenic or oncogenic effectors like polymerase beta and Ref-1 in a setting of increased hepatocyte proliferation and decreased apoptosis may facilitate peroxisome proliferator-induced hepatocellular carcinoma in the rat. PMID- 12565797 TI - Interleukin (IL)-8 and growth related oncogene-alpha in severe endotoxemia and the effects of a tumor necrosis factor-alpha/IL-1beta inhibitor on these chemokines. AB - FR167653 inhibits the production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, powerful inducers of CXC chemokines IL-8 and growth related oncogene (GRO)-alpha. The production of IL-8 and GRO-alpha was investigated and the effects of FR167653 were examined in a rabbit model of endotoxin shock. Male New Zealand rabbits were given endotoxin at a dose sufficient to induce DIC. Three groups of rabbits received FR167653 at different doses. TNF-alpha, IL-1beta, IL-8, and GRO-alpha levels were measured, several pathologic features were evaluated, and the results were compared with those obtained in control rabbits, which received only endotoxin. Endotoxin increased serum levels of IL-8 and GRO-alpha, which were associated with hypotension, renal dysfunction, and mortality, peaking at 4 h. FR167653 improved mortality, an event that was associated with decreased levels of not only TNF-alpha and IL-1beta but also IL-8 and GRO-alpha. TNF-alpha peaked at 2 h, at a time point before IL-8 and GRO-alpha reached their peak, and the TNF-alpha level was tightly correlated with that of IL-8 and GRO-alpha. Altogether, these data suggest the possible involvement of IL-8 and GRO-alpha in endotoxin shock, and FR167653 may foster a beneficial outcome in part by modulating the chemokines level by inhibiting TNF alpha and IL-1beta. PMID- 12565798 TI - The double heterozygote of two endothelin-1 gene polymorphisms (G8002A and -3A/ 4A) is related to big endothelin levels in chronic heart failure. AB - The aim of this study was to focus on the relationship among the associated genotypes of G (8002) A and -3A/-4A endothelin-1 (ET-1) gene polymorphisms and some clinical and/or biochemical parameters in Czech (Caucasian) patients with chronic heart failure. Included in the study were 103 patients with chronic heart failure (functional classes NYHA II-IV, ejection fraction < 40%). The ET-1 gene polymorphisms were detected by polymerase chain reaction (PCR) and restriction fragment length polymorphism methods. A significant decrease in the ET-1 associated genotype AG3A4A number (double heterozygote) was observed in CHF patients with plasma big endothelin levels above 0.7 pmol/L compared to those with levels below 0.7 pmol/L (OR = 0.19; 95% confidence interval = 0.06-0.57; P = 0.005; Pcorr = 0.03). We found a significant decrease in the AG3A4A genotype number in the other groups compared to the group of patients with both big endothelin and endothelin-1 levels under 0.7 pmol/L (OR = 0.22; 95% confidence interval = 0.07-0.79; P = 0.02). The double heterozygote variants of two ET-1 gene polymorphisms were associated with significantly less risk for chronic heart failure with higher levels of big endothelin. PMID- 12565799 TI - Increase in DNA polymerase gamma in the hearts of adriamycin-administered rats. AB - It is hypothesized that the cause of myocardiopathy is oxidative damage to mitochondrial DNA. To clarify this hypothesis, DNA polymerase gamma activity, which is related to the final step of mitochondrial DNA repair or renewal, was measured. One cycle of treatment consisted of five injections of adriamycin over 5 days at a dose of 1 mg/kg of body weight per day and then 2 days resting time. DNA polymerase gamma activities in the heart after one cycle of treatment were lower than the control level. However, DNA polymerase gamma activities increased with continued adriamycin treatment, reaching a maximum level in the heart at 14 days after two cycles of adriamycin treatment. Induction of DNA polymerase gamma activity was found in rat heart following three and four cycles of administration. Under these conditions, it is doubtful that mitochondrial DNA is the direct target of adriamycin administration. The damaged mitochondrial DNA may be protected by actions of the renewal or repair systems, maintaining mitochondrial function in the heart. Rat hearts at 7 days after one cycle of adriamycin treatment show morphological changes in the mitochondria that include matrix swelling and cristae disorganization, as seen in cardiac cells by electron microscopy; however, 28 days after treatment, the mitochondria appear to have recovered. PMID- 12565800 TI - Role of aquaporin water channels in eye function. AB - The aquaporins (AQPs) are a family of more than 10 homologous water transporting proteins expressed in many mammalian epithelia and endothelia. At least five AQPs are expressed in the eye: AQP0 (MIP) in lens fiber, AQP1 in cornea endothelium, ciliary and lens epithelia and trabecular meshwork, AQP3 in conjunctiva, AQP4 in ciliary epithelium and retinal Muller cells, and AQP5 in corneal and lacrimal gland epithelia. This cell-specific expression pattern suggests involvement of AQPs in corneal and lens transparency, intraocular pressure (IOP) regulation, retinal signal transduction, and tear secretion. Indeed, humans with mutant AQP0 develop cataracts. Mice lacking AQP1 have reduced IOP and impaired corneal transparency after swelling, and mice lacking AQP4 have reduced light-evoked potentials by electroretinography. There is evidence for impaired cellular processing of AQP5 in lacrimal glands of humans with Sjogren's syndrome. AQPs facilitate fluid secretion and absorption in the eye, and hence are involved in the regulation of pressure, volume and tissue hydration. Pharmacological alteration of AQP function may provide a new approach for therapy of glaucoma, corneal edema, and other diseases of the eye associated with abnormalities in IOP or tissue hydration. PMID- 12565801 TI - Alpha-crystallin. AB - Alpha A and alpha B-crystallins are a major protein component of the mammalian eye lens. Being a member of the small heat-shock protein family they possess chaperone-like function. The alpha-crystallins and especially alpha B is also found outside the lens having an extensive tissue distribution. Alpha B crystallin is found to be over-expressed in many neurological diseases, and mutations in alpha A or B-crystallin can cause cataract and myopathy. This review deals with some of the unique properties of the alpha-crystallins emphasizing especially what we don't know about its function and structure. PMID- 12565802 TI - Reversal of protein S-glutathiolation by glutaredoxin in the retinal pigment epithelium. AB - Protein cysteines can serve both sensory and activation roles in the regulation of protein function. The modulation of mixed disulfides with glutathione may promise to be a broad mechanism of redox signalling. Using both protein extract and intact RPE cells, we have generated covalent adduction of glutathione to protein cysteines and further show that glutaredoxin (Grx-1) is able to remove glutathione from protein S-glutathiolated substrates. Our data demonstrate that glutathione can modify a wide range of RPE proteins in intact cells, but that the reversal of this process--deglutathiolation and thiol bond restoration--may require a specific catalytic reaction with glutaredoxin. More generally, our experiments support the hypothesis that glutathione can non-specifically become adducted to protein cysteines during oxidative stress, but that the specific, functional reconstitution of protein thiols depends on recognition by an oxidoreductase such as glutaredoxin. This concept offers the idea that redox signalling involves both adduction of a non-specific non-protein reducing equivalent such as glutathione and specific protein based removal by glutaredoxin. PMID- 12565803 TI - Local suppression of IL-1 by receptor antagonist in the rat model of corneal alkali injury. AB - Proinflammatory cytokines, including interleukin-1 (IL-1) have been implicated in the inflammation that follows corneal alkali injury. The purpose of this series of experiments was to test whether topically applied interleukin-1 receptor antagonist (IL-1ra) could suppress corneal inflammation and promote transparency after alkali injury. Alkali injury was induced on day 0 by application of 1N NaOH to both eyes of Wistar rats (n = 28). Immediately thereafter, eyes received either topical IL-1ra (20 mg ml(-1)) in 0.2% sodium hyaluronate or vehicle alone three times daily during days 0-14. Biomicroscopic features including corneal opacity and neovascularization were assessed using a standard grading scheme. Inflammation was quantified histologically. Corneas excised at day 3 and 7 (randomly selected six eyes in each group per time point studied) were homogenized, and levels of IL-1alpha, IL-1beta, IL-6, IL-8, IL-10, and RANTES were quantified by enzyme-linked immunosorbent assay. Epithelial wound healing was examined by computed analysis of fluorescein stained corneal photographs taken daily until day 14. According to these evaluations, eyes treated with IL 1ra maintained corneal transparency with minimal neovascular invasion. Additionally, corneal damage and cell infiltration were reduced on day 7 (infiltration cells were almost 40% decreased). All cytokine/chemokine levels in IL-1ra treated eyes were significantly lower at day 3, and IL-6 and IL-10 remained significantly lower at day 7 compared to vehicle-treated eyes. IL-1ra treatment retarded epithelial wound healing in the early stage (day 1-4); however, subsequently IL-1ra treated eyes had enhanced healing with full epithelial closure at nearly the same time point as vehicle-treated eyes (day 10). We conclude that local antagonism of IL-1 after alkali injury can significantly decrease corneal inflammation and lead to enhanced corneal transparency. PMID- 12565804 TI - Delay of cataract development in hereditary cataract UPL rats by disulfiram and aminoguanidine. AB - The UPL rat is a newly developed hereditary cataract model. We previously found that the administration of disulfiram, a dimer of diethyldithiocarbamate that possesses antioxidant activity, and aminoguanidine, which is known to inhibit inducible nitric oxide synthase, inhibits cataract development in selenite induced cataract rats. In this study, we investigated the anti-cataract effects and mechanism of disulfiram and aminoguanidine on UPL rats. The opacities of UPL rat lenses, as documented by the anterior eye segment analysis system, EAS-1000 (Nidek, Aichi, Japan), increased from 39 days, and apparently mature cataracts were observed at 53 days. Accompanied with the increase in lens opacity, glutathione concentrations in UPL rat lenses decreased. The Na(+) to K(+) and water-insoluble to water-soluble protein ratios, as well as the Ca(2+) contents in UPL rat lenses increased with the development of cataracts. Oral administration of disulfiram and aminoguanidine delayed the lens opacification as well as the changes in glutathione, Na(+) to K(+) ratio, water-insoluble to soluble protein ratio, and Ca(2+) content in UPL rat lenses. The opacity and Ca(2+) content of UPL rat lenses were closely associated. The present study demonstrates that disulfiram and aminoguanidine have potency of the delay of cataract development in UPL rats, probably caused by inhibiting the rise in Ca(2+) levels. PMID- 12565805 TI - Decreased caspase-3 activity in human lens epithelium from posterior subcapsular cataracts. AB - Apoptosis has been implied in normal lens development in the embryo as well as in lens fibre differentiation. It has also been suggested to play a role in non congenital cataract and in the formation of posterior subcapsular opacification, but data on the presence of apoptosis in human lens epithelium from cataractous lenses are scarce and conflicting. The present study aimed to investigate apoptosis in lens epithelium from patients undergoing cataract surgery. The amount of apoptosis detected was correlated to age, gender, type of cataract, medications and disease. Moreover, the ability of human lens epithelial cells in culture to respond to the apoptosis-inducing agent staurosporin by activation of caspase-3 was investigated. Human lens capsulotomy specimens were collected immediately after surgery, frozen and later analysed with respect to caspase-3 activity, using the fluorogenic substrate Ac-DEVD-AMC. Generally, the activity of caspase-3 detected in this manner was very low and in 23% of the specimens it was non-detectable. However, there were differences in caspase activity between lens epithelial cells from different types of cataract, where samples from lenses with posterior subcapsular cataract exhibited significantly lower caspase-3 activity than lenses with a clear subcapsular zone. Age, gender or medications did not show any correlation with caspase activity but human capsulotomy specimens from diabetic patients exhibited significantly lower caspase-3 activity. Staurosporin caused a concentration-dependent increase in caspase activity in cultured human lens epithelial cells and the amount of apoptotic nuclei was also increased as viewed by staining with Hoechst 33342, showing chromatin condensation and nuclear fragmentation. Similar results were obtained when fresh human lens capsulotomy specimens were exposed to 1000 nM staurosporin for 24 hr. To conclude, the present data indicate that human lens epithelial cells have the ability to respond to apoptosis-inducing agents with caspase-3 dependent apoptosis, and that even though the general level of apoptosis in human lens epithelium in vivo is low, there are differences in caspase-3 activity levels in lenses with or without posterior subcapsular cataract. The latter finding supports previous studies indicating that this type of cataract may result from defective differentiation, in which apoptosis may play an important role. PMID- 12565806 TI - Utilization of lacrimal urea assay in the monitoring of hemodialysis: conditions, limitations and lacrimal arginase characterization. AB - The lacrimal urea content was found to be proportional to that of blood, which suggested its possible utilization in the monitoring of hemodialysis as a less invasive method. On the other hand, however, arginase activity was detected in tears, which may influence the urea content independently of blood urea concentration. The feasibility of using lacrimal urea measurement to replace blood urea measurement in the monitoring hemodialysis was also investigated. Blood and tear samples of 35 healthy persons and 43 renal patients undergoing hemodialysis were tested. Tear samples were collected on Schirmer paper strips. After elution the lacrimal urea content was measured by a colorimetric method. The determination of arginase activity was based on the release of urea and ornithine. The correlation between blood and lacrimal urea and arginase was studied by multivariate regression analysis. The lacrimal arginase isoenzyme pattern was investigated by native polyacrylamide gel electrophoresis and Western blotting. The effect of partially isoform-specific inhibitors was also studied. Blood urea levels in blood were significantly higher in the renal patients before dialysis than in the control patients (12.86 +/- 0.59 vs. 6.45 +/- 0.41 mM, p < 0.0001). Blood sera arginase activity was very low. Lacrimal arginase activity was significantly higher in tears than in sera (p < 0.0001 for each group). The tear/serum ratio of urea content was significantly different between controls and renal patients, particularly in postdialytic samples (1.89 +/- 0.07 vs. 3.49 +/- 0.31, p < 0.0001). The correlation between lacrimal and blood sera urea was best in controls (r = 0.89) and was better in predialytic (r = 0.75) than in postdialytic (r = 0.52) samples, depending on the level of arginase activity. In postdialytic samples a stronger correlation (r = 0.77) between tear urea and arginase was observed. Both arginase isoforms were detected in tears, but the extrahepatic (arginase II) isoenzyme was present in higher concentration. In conclusion, the determination of lacrimal urea level as a possible less invasive replacement for blood urea determination could only be utilized in the monitoring of hemodialysis if lacrimal arginase is also measured. Blood urea levels can be correctly determined by using equations, which take into account arginase activity. The accuracy of these equations was checked on a new patient population. Both arginase isoenzymes were observed in lacrimal samples. PMID- 12565807 TI - Effect of ectopic epithelial tissue within the stroma on keratocyte apoptosis, mitosis, and myofibroblast transformation. AB - The purpose of this study was to examine the effects of the epithelium on processes involved in stromal wound healing. Lamellar epithelial-stromal flaps were produced in rabbit corneas with a microkeratome. Peripheral corneal epithelial tissue, central corneal epithelial tissue, or no epithelial tissue (control) was introduced beneath the flap. Corneas were removed at time points from 4 hr to 1 month after surgery. Tissue sections were analyzed with immunocytochemistry for Keratin 3 (K3) to detect epithelial antigen, terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labelling (TUNEL) assay to detect apoptosis, immunocytochemistry for Ki67 to detect cell proliferation, and immunocytochemistry for alpha-smooth muscle actin (SMA) to detect myofibroblasts. K3 was detected at the level of the interface from 4 hr to 1 month after surgery in corneas in which epithelial tissue was introduced, but not control corneas, with the exception of one that developed epithelial in growth. Keratocyte apoptosis was significantly higher at 4 hr after flap formation in both groups in which corneal epithelial tissue was introduced beneath the flap compared with controls. Keratocyte proliferation was significantly greater at 72 hr in corneas in which epithelial tissue was introduced beneath the flap compared to the controls. Corneas in which epithelial tissue was introduced into the interface, but not control corneas, had stromal cells expressing alpha-SMA in the stroma anterior and posterior to the interface at 1 week and 1 month after surgery. This was also noted in the control cornea in which there was epithelial ingrowth. Signals derived from the corneal epithelium promote keratocyte apoptosis. Keratocyte proliferation is higher in corneas that have lamellar surgery when epithelial tissue is introduced into the interface. Epithelium-derived signals also participate in the generation and/or maintenance of myofibroblasts in the corneal stroma. PMID- 12565809 TI - Edible mushroom (Agaricus bisporus) lectin modulates human retinal pigment epithelial cell behaviour in vitro. AB - The retinal pigment epithelium (RPE) plays a major role in the development of proliferative vitreoretinopathy (PVR). In particular, RPE cells are implicated in generating the contraction forces seen. The present study was undertaken to investigate whether human RPE binds a lectin from the common edible mushroom, Agaricus bisporus, and to evaluate the effect of any binding on RPE-mediated matrix contraction in an in vitro model of PVR. Fluorescein isothiocyanate (FITC) labelled Agaricus bisporus lectin (ABL) was used to study binding of lectin to normal retina, PVR scar tissue specimens and cultured human RPE. The effect of a 3-day exposure of ABL on human RPE-mediated contraction was evaluated using 2- and 3D RPE-populated collagen matrices. Effect of ABL on cell adhesion was measured using a collagen type I adhesion assay and determining the relative cellular attachment using absorbance readings. The normal RPE monolayer did not stain with FITC-ABL while PVR scar tissue stained intensely. Staining of in vitro RPE was characteristic but time-dependent. ABL caused a dose-dependent inhibition of RPE-mediated contraction of both 2D (one-way ANOVA, F = 7.94, p < 0.008) and 3D collagen matrices (one-way ANOVA, F = 164.955, p < 0.001). Pre-incubation of ABL with RPE in the 2D model caused a dramatic arrest of contraction (one-way ANOVA, F = 20.1, p < 0.001) that was due to a dose-dependent inhibition of adhesion (one-way ANOVA, F = 15.603, p < 0.001). Recovery of contraction was partially reversible on removal of ABL and was dependent on initial concentration of the lectin. ABL inhibits contraction and adhesion of human RPE cells in vitro without apparent cytotoxicity. It therefore deserves consideration as a potential therapeutic agent in the prevention and treatment of PVR and other non-ocular anomalous wound-healing processes. PMID- 12565808 TI - In vivo micropathology of Best macular dystrophy with optical coherence tomography. AB - Best macular dystrophy (BMD) is an autosomal dominant retinopathy caused by mutations in the VMD2 gene that encodes a chloride channel in the basolateral membrane of the retinal pigment epithelium (RPE). BMD patients were studied using optical coherence tomography (OCT) to understand the disease process in the macula leading to vision loss. BMD patients (ages 5-61), representing four families with known VMD2 mutations, were included. OCT scans were recorded in the central retina and longitudinal reflectivity profiles were analysed. The central retina in BMD showed different OCT abnormalities at or near the level of the highly reflective deep retinal band termed the outer retina-choroid complex (ORCC). Two types of ORCC change were noted to occur either separately or together: (1) splitting with or without intervening hyporeflective areas; and (2) elevation. Longitudinal study of a BMD patient indicated that such abnormalities were dynamic and changed in type and degree with time. The pathogenetic sequence in BMD may begin with defective fluid transport across the RPE secondary to the channelopathy in the basolateral membrane. In the macula, this leads to an abnormal interface with adjacent structures at both apical and basal surfaces of the RPE. The disease process results in detachments of the neurosensory retina, such as in central serous chorioretinopathy, and sub-RPE pathology resembling some stages of age-related macular degeneration, with eventual loss of photoreceptors, inner retina and central vision. PMID- 12565810 TI - Role and regulation of CXC-chemokines in acute experimental keratitis. AB - The aim of this study was to elucidate the expression of chemokines, their role and regulation in bacterial corneal infection using three bacterial strains (Pseudomonas. aeruginosa- invasive, cytotoxic and contact lens induced acute red eye strains) which have been shown to produce three distinct patterns of corneal disease in the mouse. The predominant chemokine expressed in response to all three strains was MIP-2. Prolonged expression of high levels of MIP-2 was associated with increased severity of corneal inflammation. Significantly reduced disease severity upon administration of anti-MIP-2 antibodies suggested that MIP 2 may play an important role in the pathogenesis of Pseudomonas keratitis at least in part by being a major chemoattractant for polymorphonuclear leukocytes (PMN) recruitment. Interestingly, the numbers of bacteria in eyes with neutralized MIP-2 activity did not decrease even though the severity of the disease was decreased. This implies PMNs as the major destructive factor in microbial keratitis. Further, neutralization of IL-1beta activity alone using monoclonal antibodies resulted in significant reduction of both MIP-2 and KC activity indicating that chemokine levels were regulated by IL-1beta. These studies demonstrate that the regulation of MIP-2 activity may be beneficial in reducing corneal damage during microbial keratitis in rodents and perhaps that regulation of the human homologue of MIP-2, IL-8, may be useful for controlling keratitis in humans. PMID- 12565811 TI - Quantitative analysis of lacrimal gland function, apoptotic figures, Fas and Fas ligand expression of lacrimal glands in dry eye patients. AB - The purpose of our study was to examine the correlation between the lacrimal gland function and apoptotic figure, Fas and Fas ligand (FasL) expression in the lacrimal gland. A total of 15 dry eye patients (nine Sjogren's syndrome and six non-Sjogren's syndrome-type dry eye) were recruited for the study. Lacrimal function was evaluated by Schirmer tests 1 and 2. Lacrimal gland biopsies were performed and sections were analyzed by immunohistochemistry using APO2.7, an antibody to Fas and FasL. Quantitative analysis of fluorescein staining was performed by a scanning laser microscopy. Schirmer test 2 results were lower in Sjogren's syndrome-type dry eye and were associated with positive staining of acinar cells with APO2.7 and of infiltrating lymphocytes with FasL. There was a good correlation between the results of Schirmer test 2 and APO2.7 and FasL staining. Lacrimal gland dysfunction is related to the apoptotic figure of acinar cells possibly induced by FasL on the infiltrating lymphocytes. PMID- 12565812 TI - Alpha lipoic acid changes iron uptake and storage in lens epithelial cells. AB - Alpha lipoic acid (LA) is a cofactor in mitochondrial dehydrogenase complexes. Previous studies have shown that when administered exogenously LA has antioxidant properties, which include free radical scavenging, metal chelation and regeneration of other antioxidants. The cells convert LA into dihydroplipoic acid (DHLA), which in the presence of iron can act as a prooxidant. In vitro DHLA reduces Fe(+3) to Fe(+2) and removes iron from ferritin, increasing the risk of Fe catalyzed free radical formation. In the present study we examined the in vivo effects of lipoic acid treatment on Fe metabolism in cultured lens epithelial cells, and found that LA decreases Fe uptake from transferrin, increases Fe deposition into ferritin and increases the concentration of this protein. When administered together with ascorbic acid, lipoic acid changes the characteristic heavy to light chain ratio of ferritin makeup. The decreased Fe uptake and increased storage diminishes the size of the cytosolic highly reactive Fe pool (LIP). These changes are associated with increased cell resistance to H(2)O(2) challenge. Therefore, LA may reduce the risk of Fe induced oxidative damage and also might be useful as a treatment of Fe overload. PMID- 12565813 TI - Human RPE cells express the FGFR2IIIc and FGFR3IIIc splice variants and FGF9 as a potential high affinity ligand. AB - The expression of splice variants of FGF receptors, which differ in the third Ig domain, was investigated in retinal pigment epithelium (RPE) cells in vitro and in vivo. This region of the protein determines ligand-binding specificity. Additionally, the expression of potential ligands for these receptors was investigated. Expression of FGF receptor transcript alternative splicing was analyzed by RT-PCR/Southern analysis in RPE cells in vitro and in vivo. The expression of FGFs by RT-PCR, in situ hybridization, and immunohistochemistry in sections of the human posterior pole was also investigated. The ARPE-19 cell line expresses only the FGFR2IIIc splice variant and does not express any FGFR3 splice variants in vitro. Two in vivo samples exhibited expression of the FGFR2IIIc and FGFR3IIIc splice variants and no evidence of the corresponding IIIb splice variant. The results from previous studies for these receptors imply that FGF9 or FGF4 could act as ligands. We demonstrated that FGF9 is expressed in a subpopulation of the RPE, as well as photoreceptors and other neurons of the retina. FGF4 was not detected by RT-PCR analysis in RPE cells in vitro. These data suggest that FGF9 may be an autocrine/paracrine factor in the outer retina. PMID- 12565814 TI - CRALBP transcriptional regulation in ciliary epithelial, retinal Muller and retinal pigment epithelial cells. AB - Cellular retinaldehyde binding protein (CRALBP) functions in the visual cycle and mutations in the RLBP1 gene can lead to blindness. RLBP1 promoter analyses have been pursued in vitro as an approach to deciphering the mechanism controlling cell-specific expression of CRALBP. Reporter activity of wildtype and mutant RLBP1 promoter constructs suggest that CRALBP transcriptional regulation may be similar in the ciliary epithelium (CE) and retinal pigment epithelium (RPE) but different in Muller cells. Results in RPE cells refine the location of an RLBP1 enhancer element to within -1826 to -1749 bp and a repressor element to within 702 to -635 bp. PMID- 12565815 TI - Necrosis: a specific form of programmed cell death? AB - For a long time necrosis was considered as an alternative to programmed cell death, apoptosis. Indeed, necrosis has distinct morphological features and it is accompanied by rapid permeabilization of plasma membrane. However, recent data indicate that, in contrast to necrosis caused by very extreme conditions, there are many examples when this form of cell death may be a normal physiological and regulated (programmed) event. Various stimuli (e.g., cytokines, ischemia, heat, irradiation, pathogens) can cause both apoptosis and necrosis in the same cell population. Furthermore, signaling pathways, such as death receptors, kinase cascades, and mitochondria, participate in both processes, and by modulating these pathways, it is possible to switch between apoptosis and necrosis. Moreover, antiapoptotic mechanisms (e.g., Bcl-2/Bcl-x proteins, heat shock proteins) are equally effective in protection against apoptosis and necrosis. Therefore, necrosis, along with apoptosis, appears to be a specific form of execution phase of programmed cell death, and there are several examples of necrosis during embryogenesis, a normal tissue renewal, and immune response. However, the consequences of necrotic and apoptotic cell death for a whole organism are quite different. In the case of necrosis, cytosolic constituents that spill into extracellular space through damaged plasma membrane may provoke inflammatory response; during apoptosis these products are safely isolated by membranes and then are consumed by macrophages. The inflammatory response caused by necrosis, however, may have obvious adaptive significance (i.e., emergence of a strong immune response) under some pathological conditions (such as cancer and infection). On the other hand, disturbance of a fine balance between necrosis and apoptosis may be a key element in development of some diseases. PMID- 12565816 TI - Mechanisms of c-myc-mediated transcriptional repression of growth arrest genes. AB - Constitutive expression of the proto-oncogene c-myc results in oncogenic activation and contributes to progression of a wide range of human and animal tumors. Myc executes its multiple activities mostly through transcriptional regulation of the target genes. The special interest of this review is the mechanism of transcriptional repression of cell cycle inhibitors by Myc. Myc suppresses expression of cell cycle/growth arrest genes gas1, p15, p21, p27, and gadd34, -45, and -153. It appears that Myc represses growth arrest gene transcription by at least two distinct mechanisms. One mechanism is limited to the binding of Myc-Max heterodimers to the Inr element in their promoters and inhibition of Miz-1 or other transcriptional activators via the C-terminal domain of c-Myc. This mechanism requires DNA binding of the Myc-Max complex to Inr sequences. The other mechanism is dependent on c-Myc binding to the Sp1 transcription factor via the c-Myc central region and inhibition of Sp1 transcriptional activity. At this time it is not entirely clear which Sp1 containing promoters will be repressed by c-Myc and what other modes of c-Myc transcriptional repression may exist. The ability of c-Myc to repress transcription of growth arrest genes may contribute to its potential to promote proliferation and oncogenesis. PMID- 12565817 TI - An analysis of replicative senescence in dermal fibroblasts derived from chronic leg wounds predicts that telomerase therapy would fail to reverse their disease specific cellular and proteolytic phenotype. AB - The accumulation of senescent fibroblasts within tissues has been suggested to play an important role in mediating impaired dermal wound healing, which is a major clinical problem in the aged population. The concept that replicative senescence in wound fibroblasts results in reduced proliferation and the failure of refractory wounds to respond to treatment has therefore been proposed. However, in the chronic wounds of aged patients the precise relationship between the observed alteration in cellular responses with aging and replicative senescence remains to be determined. Using assays to assess cellular proliferation, senescence-associated staining beta-galactosidase, telomere length, and extracellular matrix reorganizational ability, chronic wound fibroblasts demonstrated no evidence of senescence. Furthermore, analysis of in vitro senesced fibroblasts demonstrated cellular responses that were distinct and, in many cases, diametrically opposed from those exhibited by chronic wound fibroblasts. Forced expression of telomerase within senescent fibroblasts reversed the senescent cellular phenotype, inhibiting extracellular matrix reorganizational ability, attachment, and matrix metalloproteinase production and thus produced cells with impaired key wound healing properties. It would appear therefore that the distinct phenotype of chronic wound fibroblasts is not simply due to the aging process, mediated through replicative senescence, but instead reflects disease-specific cellular alterations of the fibroblasts themselves. PMID- 12565818 TI - The homeodomain-interacting kinase PKM (HIPK-2) modifies ND10 through both its kinase domain and a SUMO-1 interaction motif and alters the posttranslational modification of PML. AB - Homeodomain-interacting protein kinases (HIPK-1, -2, and -3) are a family of enzymes that have been implicated in the phosphorylation and repression of homeodomain-containing transcription factors. HIPK-2 has been found to interact with the SUMO-1-conjugating enzyme Ubc9 and can be covalently modified by SUMO-1. It has also been shown to interact with and phosphorylate p53 and to form punctate speckles in the nucleus of which a proportion colocalize with PML nuclear bodies (ND10). We have previously shown that the hamster equivalent of HIPK-2 (named PKM) interacts with the interferon-induced antiviral GTPase Mx1 and associates with ND10 in interferon-treated cells. Given the connections between the interferon response pathway, constituents of ND10, and SUMO-1-conjugated proteins, we have studied the effects of exogenously expressed PKM on endogenous ND10 proteins. We found that PKM induces structural changes in ND10 that can be attributed both to its kinase activity and to the presence of a functional SUMO-1 interaction motif in the C-terminal half of the protein. The changes in the localization of PML, Sp100, and hDaxx induced by exogenous PKM or fragments thereof correlate with changes in the posttranslationally modified species of PML. We propose that PKM is able to modify ND10 structure by inducing changes in the posttranslational modification of PML and by interacting with SUMO-1 modification pathways. PMID- 12565819 TI - Apoptosis by Par-4 in cancer and neurodegenerative diseases. AB - Prostate apoptosis response-4 (par-4) is a pro-apoptotic gene identified in prostate cancer cells undergoing apoptosis. Par-4 protein, which contains a leucine zipper domain at the carboxy-terminus, functions as a transcriptional repressor in the nucleus. Par-4 selectively induces apoptosis in androgen independent prostate cancer cells and Ras-transformed cells but not in androgen dependent prostate cancer cells or normal cells. Cells that are resistant to apoptosis by Par-4 alone, however, are greatly sensitized by Par-4 to the action of other pro-apoptotic insults such as growth factor withdrawal, tumor necrosis factor, ionizing radiation, intracellular calcium elevation, or those involved in neurodegenerative diseases such as Alzheimer's, Parkinson's, Huntington's, and stroke. Apoptosis induction by Par-4 involves a complex mechanism that requires activation of the Fas death receptor signaling pathway and coparallel inhibition of cell survival NF-kappaB transcription activity. The unique ability of Par-4 to induce apoptosis in cancer cells but not normal cells and the ability of Par-4 antisense or dominant-negative mutant to abrogate apoptosis in neurodegenerative disease paradigms makes it an appealing candidate for molecular therapy of cancer and neuronal diseases. PMID- 12565820 TI - The role of alpha3beta1 integrin in determining the supramolecular organization of laminin-5 in the extracellular matrix of keratinocytes. AB - Analyses of mice with targeted deletions in the genes for alpha3 and beta1 integrin suggest that the alpha3beta1 integrin heterodimer likely determines the organization of the extracellular matrix within the basement membrane of skin. Here we tested this hypothesis using keratinocytes derived from alpha3 integrin null mice. We have compared the organizational state of laminin-5, a ligand of alpha3beta1 integrin, in the matrix of wild-type keratinocytes with that of laminin-5 in the matrix of alpha3 integrin-null cells. Laminin-5 distributes diffusely in arc structures in the matrix of wild-type mouse keratinocytes, whereas laminin-5 is organized into linear, spike-like arrays by the alpha3 integrin-null cells. The fact that alpha3 integrin-null cells are deficient in their ability to assemble a proper laminin-5 matrix is also shown by their failure to remodel laminin-5 when plated onto surfaces coated with purified laminin-5 protein. In sharp contrast, wild-type keratinocytes organize exogenously added laminin-5 into discrete ring-like organizations. These findings led us next to assess whether differences in laminin-5 organization in the matrix of the wild-type and alpha3 integrin-null cells impact cell behavior. Our results indicate that alpha3 integrin-null cells are more motile than their wild-type counterparts and leave extensive trails of laminin-5 over the surface on which they move. Moreover, HEK 293 cells migrate significantly more on the laminin-5 rich matrix derived from the alpha3 integrin-null cells than on the wild-type keratinocyte laminin-5 matrix. In addition, alpha3 integrin-null cells show low strength of adhesion to surfaces coated with purified laminin-5 compared to wild type cells although both the wild type and the alpha3 integrin-null keratinocytes adhere equally strongly to laminin-5 that has been organized into arrays by other epithelial cells. These data suggest: (1) that alpha3beta1 integrin plays an important role in determining the incorporation of laminin-5 into its proper higher-order structure within the extracellular matrix of keratinocytes and (2) that the organizational state of laminin-5 has an influence on laminin-5 matrix function. PMID- 12565821 TI - The kinase domain of MEKK1 induces apoptosis by dysregulation of MAP kinase pathways. AB - MAP kinase pathways comprise a group of parallel protein phosphorylation cascades, which are involved in signaling triggered by a variety of stimuli. Previous findings suggested that the ERK and the JNK pathways have opposing roles in regulating proliferation and survival or apoptosis and that apoptosis can be promoted by inhibiting the ERK pathway or by activation of the JNK pathway. In order to test this hypothesis and explore whether it can be exploited as a strategy for killing human cancer cells, we used gene transfer experiments with a range of cancer cell lines. We expressed the catalytic fragment of human MEKK1 to activate JNK and the Ras-binding domain (RBD) of Raf-1 to inhibit the Ras-ERK pathway. In addition, we designed several RBD-MEKK1 fusion proteins aiming to simultaneously activate the JNK and block the ERK pathway. We found that the MEKK1 proteins as well as the RBD alone could reduce colony formation in all cell lines. The survival time of MEKK1-expressing cells depended on the cell line. In HeLa cells, survival could be prolonged by inhibition of caspases but not by coexpression of the anti-apoptotic protein Bcl-2. Due to a lower kinase activity the RBD-MEKK1 fusion proteins were less effective in apoptosis induction than the MEKK1 kinase domain alone. Using mutant forms of Ras and Raf-1 we could show that the reduced kinase activity of RBD-MEKK1 fusion proteins was caused by binding to the Ras protein. The expression of lethal doses of MEKK1 resulted in a strong activation of all three major MAP kinase families JNK, ERK, and p38. Blocking these pathways either by coexpressing a dominant negative form of MKK4 or with inhibitors of MEK or p38 failed to inhibit apoptosis. This suggests that MEKK1 induces apoptosis by causing a general deregulation of MAP kinase signaling rather than by the activation of a single pathway. PMID- 12565822 TI - A permeable FGF-1 nuclear localization sequence peptide induces DNA synthesis independently of Ras activation. AB - A 26-amino-acid peptide (designated PFNP) composed of the nuclear localization signal of fibroblast growth factor (FGF)-1 and a membrane-permeable peptide is known to mimic FGF-1's ability to stimulate DNA synthesis in various cell types at low cell densities. The underlying molecular mechanism is unknown, however. Here we show that PFNP activity is inhibited in murine fibroblasts by a tyrosine kinase inhibitor, that PFNP does not bind to the FGF receptor, and that PFNP does not induce phosphorylation of the FGF receptor substrate. In addition, expression of a dominant-negative form of Ras, which abolished the activities of epidermal growth factor (EGF) and heparin-binding EGF, had no affect on PFNP-induced DNA synthesis. Despite this apparent Ras independence, PFNP activity correlated with phosphorylation of ERK1/2 MAP kinases and was concentration dependently inhibited by inhibitors of ERK1/2 MAP kinase phosphorylation. These results indicate that whereas Ras activation is dispensable for PFNP-induced DNA synthesis, activation of tyrosine kinases and ERK1/2 kinases, albeit independently of the FGF receptor system, is crucial. Interestingly, FGF-1 signaling was predominantly Ras independent when the cell density was optimum for PFNP, suggesting that PFNP and FGF-1 share the same signaling mechanism. PMID- 12565823 TI - The kic1 kinase of schizosaccharomyces pombe is a CLK/STY orthologue that regulates cell-cell separation. AB - The CLK/STY kinases are a family of dual-specificity protein kinases implicated in the regulation of cellular growth and differentiation. Some of the kinases in the family are shown to phosphorylate serine-arginine-rich splicing factors and to regulate pre-mRNA splicing. However, the actual cellular mechanism that regulates cell growth, differentiation, and development by CLK/STY remains unclear. Here we show that a functionally conserved CLK/STY kinase exists in Schizosaccharomyces pombe, and this orthologue, called Kic1, regulates the cell surface and septum formation as well as a late step in cytokinesis. The Kic1 protein is modified in vivo, likely by phosphorylation, suggesting that it can be involved in a control cascade. In addition, kic1(+) together with dsk1(+), which encodes a related SR-specific protein kinase, constitutes a critical in vivo function for cell growth. The results provide the first in vivo evidence for the functional conservation of the CLK/STY family through evolution from fission yeast to mammals. Furthermore, since cell division and cell-cell interaction are fundamental for the differentiation and development of an organism, the novel cellular role of kic1(+) revealed from this study offers a clue to the understanding of its counterparts in higher eukaryotes. PMID- 12565824 TI - The interplay between integrins alphaMbeta2 and alpha5beta1 during cell migration to fibronectin. AB - A directed migration of leukocytes through the extracellular matrix requires the regulated engagement of integrin cell adhesion receptors. The integrin alpha(M)beta(2) (CD11b/CD18, Mac-1) is progressively upregulated to high levels on migrating phagocytic leukocytes in response to inflammatory stimuli and is able to bind numerous ligands in the interstitial matrix. The role of alpha(M)beta(2) in migration of leukocytes through the extracellular matrix and its cooperation with other leukocyte integrins during migration are not understood. Using a model system consisting of cells that express different levels of alpha(M)beta(2) and an invariable level of endogenous integrin alpha(5)beta(1), we have explored a situation relevant to migrating neutrophils when alpha(M)beta(2) and alpha(5)beta(1) engage the same ligand, fibronectin. We show that fibronectin is a ligand for alpha(M)beta(2) and that both alpha(M)beta(2) and alpha(5)beta(1) on the alpha(M)beta(2)-expressing cells contribute to adhesion to fibronectin. However, migration of these cells to fibronectin is mediated by alpha(5)beta(1), whereas alpha(M)beta(2) retards migration. The decrease in migration correlates directly with the increased alpha(M)beta(2) density. Ligation of alpha(M)beta(2) with function-blocking antibodies can reverse this effect. The restorative effects of antibodies are caused by the removal of restraint imposed by the excess of alpha(M)beta(2) fibronectin adhesive bonds. These findings indicate that alpha(M)beta(2) can increase general cell adhesiveness which results in braking of cell migration mediated by integrin alpha(5)beta(1). Because alpha(M)beta(2) binds numerous proteins in the extracellular matrix with a specificity overlapping that of the beta(1) integrins, the results suggest that alpha(M)beta(2) can affect the beta(1) integrin-mediated cell migration. PMID- 12565825 TI - A human TERT C-terminal polypeptide sensitizes HeLa cells to H2O2-induced senescence without affecting telomerase enzymatic activity. AB - We have constructed a 27-kDa hTERT C-terminal polypeptide (hTERTC27) devoid of domains required for telomerase activity and demonstrated that it is capable of nuclear translocation/telomere-end targeting. Here we showed that expression of a low level of hTERTC27 renders hTERT positive HeLa cells sensitive to H(2)O(2) induced oxidative stress and subsequent cell senescence. The senescence associated gene, the cyclin/cdk inhibitor p21(Waf1), was up-regulated. This occurs without changing the expression of endogenous hTERT, causing significant telomere shortening or inhibiting telomerase activity. Results from this study suggest for the first time that in addition to telomerase activity, the C terminus of hTERT also plays a role in hTERT-mediated cellular resistance to oxidative stress. PMID- 12565826 TI - Global insights into protein complexes through integrated analysis of the reliable interactome and knockout lethality. AB - We performed an integrated computational analysis of data derived from a comprehensive set of protein-protein interactions (interactome) and a phenotype dataset on lethality in Saccharomyces cerevisiae. For the analysis, we selected reliable interactome data using our previous 'interaction generality,' a computational approach to assess reliability of interactions. Those efforts gave clear evidence that proteins with lethal phenotypes in knockout studies (lethal proteins) may interact with each other to form functional protein complexes to perform their cellular roles. However, our analysis indicates that interactions between lethal proteins are rather restricted to the same cellular pathway or function, and it is quite unlikely that they interact with other lethal proteins functioning in different cellular roles. Furthermore, our results allowed us predictions on the functions of thus far uncharacterized lethal proteins with an estimated 93% accuracy. Thus, the analysis described in here can provide global insights into the biological features of the protein complexes. PMID- 12565827 TI - Fission yeast synaptobrevin is involved in cytokinesis and cell elongation. AB - Synaptobrevin is a vesicle-associated membrane protein playing an essential role in regulated vesicle transport. In this study, we characterized Syb1, synaptobrevin of Schizosaccharomyces pombe. Syb1 was located on various sizes of vesicle-like structures in the cytoplasm and enriched in the medial region and cell ends. Transport of Syb1 to the medial region was mainly dependent on F-actin and Myo52/Myo4. Syb1 is essential for cell viability and most of the syb1-null cells showed a round or short cylindrical form. These results suggest that Syb1 is involved in membrane trafficking of cytokinesis and cell elongation. PMID- 12565828 TI - Secophalloidin as a novel activator of skinned cardiac muscle. AB - Secophalloidin (SPH) is known to activate skinned cardiac muscle in the absence of Ca(2+). We hypothesized that SPH-induced changes in cross-bridge properties underlie muscle activation. We found that force responsiveness to orthovanadate was drastically reduced in SPH activated muscles compared to Ca(2+)-activated contraction. Moreover, SPH caused approximately 30% increase in Ca(2+) independent force in muscles where Ca(2+) sensitivity was totally destroyed by troponin I extraction with 10mM vanadate. Thus, SPH and Ca(2+) activation differ in both properties of the cross-bridge cycle and protein requirements for thin filament regulation. In addition, we tested the relationship between the activating effects SPH and EMD 57033, a Ca(2+) sensitizer that increases resting force in cardiac muscle. After maximal activation by either SPH or EMD 57033, the other compound was found to further increase force, indicating that SPH activates muscle via a novel mechanism. PMID- 12565829 TI - Protein kinase CK2 phosphorylates and activates the SR protein-specific kinase 1. AB - The serine/arginine subfamily of protein kinases has been conserved throughout evolution and its members are thought to play important roles in the regulation of multiple cellular processes. Mammalian SRPK1 has been considered as a constitutively active kinase that is predominantly expressed in testis. In the present study, recombinant GST-SRPK1 was used as substrate to identify potential protein kinase(s) in testis extracts, involved in phosphorylating and thereby regulating the activity of this enzyme. Using a panel of chromatography media, inhibition by heparin, immunoblot analysis, and phosphopeptide mapping, CK2 was determined to be the major kinase that phosphorylates SRPK1. Phosphorylation of SRPK1 by CK2 occurred mainly at Ser(51) and Ser(555) in vitro, and resulted in approximately 6-fold activation of the enzyme. These findings suggest that SRPK1 may be an important cellular target for CK2 action. PMID- 12565830 TI - Expression of polycystin-1 C-terminal fragment enhances the ATP-induced Ca2+ release in human kidney cells. AB - Polycystin-1 (PC1) is a membrane protein expressed in tubular epithelia of developing kidneys and in other ductal structures. Recent studies indicate this protein to be putatively important in regulating intracellular Ca(2+) levels in various cell types, but little evidence exists for kidney epithelial cells. Here we examined the role of the PC1 cytoplasmic tail on the activity of store operated Ca(2+) channels in human kidney epithelial HEK-293 cell line. Cells were transiently transfected with chimeric proteins containing 1-226 or 26-226 aa of the PC1 cytoplasmic tail fused to the transmembrane domain of the human Trk-A receptor: TrkPC1 wild-type and control Trk truncated peptides were expressed at comparable levels and localized at the plasma membrane. Ca(2+) measurements were performed in cells co-transfected with PC1 chimeras and the cytoplasmic Ca(2+) sensitive photoprotein aequorin, upon activation of the phosphoinositide pathway by ATP, that, via purinoceptors, is coupled to the release of Ca(2+) from intracellular stores. The expression of TrkPC1 peptide, but not of its truncated form, enhanced the ATP-evoked cytosolic Ca(2+) concentrations. When Ca(2+) assays were performed in HeLa cells characterized by Ca(2+) stores greater than those of HEK-293 cells, the histamine-evoked cytosolic Ca(2+) increase was enhanced by TrkPC1 expression, even in absence of external Ca(2+). These observations indicate that the C-terminal tail of PC1 in kidney and other epithelial cells upregulates a Ca(2+) channel activity also involved in the release of intracellular stores. PMID- 12565831 TI - Affinity purification of ARE-binding proteins identifies polyA-binding protein 1 as a potential substrate in MK2-induced mRNA stabilization. AB - An important determinant for the expression level of cytokines and proto oncogenes is the rate of degradation of their mRNAs. AU-rich sequence elements (AREs) in the 3(') untranslated regions have been found to impose rapid decay of these mRNAs. ARE-containing mRNAs can be stabilized in response to external signals which activate the p38 MAP kinase cascade including the p38 MAP kinase substrate MAPKAP kinase 2 (MK2). In an attempt to identify components downstream of MK2 in this pathway we analyzed several proteins which selectively interact with the ARE of GM-CSF mRNA. One of them, the cytoplasmic poly(A)-binding protein PABP1, co-migrated with a protein that showed prominent phosphorylation by recombinant MK2. Phosphorylation by MK2 was confirmed using PABP1 purified by affinity chromatography on poly(A) RNA. The selective interaction with an ARE containing RNA and the phosphorylation by MK2 suggest that PABP1 plays a regulatory role in ARE-dependent mRNA decay and its modulation by the p38 MAP kinase cascade. PMID- 12565832 TI - Human sensitive to apoptosis gene protein inhibits peroxynitrite-induced DNA damage. AB - Sensitive to apoptosis gene (SAG) protein, a novel zinc RING finger protein, which is redox responsive and protects mammalian cells from apoptosis, is a metal chelator and a potential reactive oxygen species scavenger, but its antioxidant properties have not been completely defined. The present study was undertaken to test the hypothesis that human SAG protects from DNA damage induced by peroxynitrite, a potent physiological inorganic toxin. The present study has shown that SAG significantly inhibits single strand breaks in supercoiled plasmid DNA induced by synthesized peroxynitrite (ONOO(-)) and 3-morpholinosydnomine N ethylcarbamide (SIN-1), a generator of peroxynitrite through the reaction between nitric oxide and superoxide anion. The formation of 8-hydroxy-2(')-deoxyguanosine in calf thymus DNA by peroxynitrite and SIN-1 was also significantly inhibited by SAG. The protective effect on peroxynitrite-mediated DNA damage was completely abolished by the reaction of SAG with N-ethylmaleimide, a chemical modification agent for the sulfhydryl group of proteins. These observations suggested that the sulfhydryl group of cysteines in SAG could react directly with peroxynitrite to prevent DNA damage. PMID- 12565833 TI - Rapid disappearance of deoxyribose-1-phosphate in platelet derived endothelial cell growth factor/thymidine phosphorylase overexpressing cells. AB - Platelet derived endothelial cell growth factor/thymidine phosphorylase (PD ECGF/TP) catalyzes the phosphorolysis of thymidine (TdR) to thymine and deoxyribose-1-phosphate (dR-1-P) and has a pro-angiogenic effect for which dR-1-P may be responsible. Using a purine nucleoside phosphorylase based assay it was found that TdR incubation did not increase dR-1-P accumulation in colon cancer cell line Colo320 and its PD-ECGF/TP transfected variant Colo320TP1. The assay was linear up to 25,000pmol dR-1-P with complete recovery of dR-1-P from cellular extracts. There was a huge discrepancy between thymine production and the measured dR-1-P level, 0.05% of the expected value for dR-1-P was found, indicating that there was a rapid disappearance of dR-1-P. However, in cellular extracts, TdR incubation increased dR-1-P, measurable by trapping, which was inhibited by a thymidine phosphorylase inhibitor. dR-1-P directly added to cellular extracts disappeared within 5-10min. In conclusion, large amounts of dR 1-P are produced by Colo320TP1 cells, which rapidly disappear thus not resulting in a net accumulation of dR-1-P in these cells. PMID- 12565834 TI - The exon 7-spliced Lck isoform in T lymphocytes: a potential regulator of p56lck signaling pathways. AB - The protein-tyrosine kinase p56lck is the product of the lck gene. It plays a pivotal role in T-lymphocyte activation and thymocyte development, as indicated by the defective immune responses of lck-/- mice. We have demonstrated that an exon 7-deleted lck mRNA is produced by alternative splicing in all human cells expressing the lck gene. We have now looked for the protein encoded by this spliced lck mRNA and attempted to determine the function of the deleted Lck protein. This paper shows that the LckDelta7 protein is present in JCaM1.6 T cells and we inferred that this isoform accounts for 15% of the total Lck proteins in the parental Jurkat T-cell line. We report that deletion of the first 51 amino-acids (exon 7) of the Lck catalytic domain greatly reduces the kinase activity of the recombinant protein. The residual activity can, nevertheless, be enhanced by adding Mn(2+), whereas this cation has no effect on the activity of the p56lck mutated in its active site (K273E). The enforced production of LckDelta7 protein in transfected Jurkat cells results in slower cell proliferation than does p56lck. These findings suggest that the LckDelta7 protein is a p56lck cell-signaling regulator. This mechanism could be common to both humans and mice, in which we also found the exon 7-spliced lck transcript. PMID- 12565835 TI - Identification and distribution of different mRNA variants produced by differential splicing in the human phosphodiesterase 9A gene. AB - The transcript population of the human gene coding for a cGMP-dependent phosphodiesterase (PDE9A) has a complex structure. There is a high level of mRNA in intestinal and prostate tissues, a low level in blood, and intermediate in other tissues. More than 20 different variants produced by differential splicing have been observed and new exons have been identified both by PCR amplification and by the analysis of available EST sequences. In all cases the transcriptional start site is the same and no differential splicing is found in the exons coding for the catalytic domain of the protein. In some cases the protein produced by splice variants is truncated. The distribution of the splice variants is not homogeneous among the different tissues studied. The human, but not the mouse, PDE9A gene appears to have a complex regulation of expression by different isoforms. PMID- 12565836 TI - Critical role of N-terminal N-glycosylation for proper folding of the human formyl peptide receptor. AB - The human formyl peptide receptor (FPR) is N-glycosylated and activates phagocytes via G(i)-proteins. The FPR expressed with G(i)alpha(2)beta(1)gamma(2) in Sf9 insect cells exhibits high constitutive activity as assessed by strong inhibitory effects of an inverse agonist and Na(+) on basal guanosine 5(')-O-(3 thiotriphosphate) (GTPgammaS) binding. The aim of our study was to analyze the role of N-glycosylation in FPR function. Site-directed mutagenesis of extracellular Asn residues prevented FPR glycosylation but not FPR expression in Sf9 membranes. However, in terms of high-affinity agonist binding, kinetics of GTPgammaS binding, number of G(i)-proteins activated, and constitutive activity, non-glycosylated FPR was much less active than native FPR. FPR Asn4Gln/Asn10Gln/Asn179Gln and FPR-Asn4Gln/Asn10/Gln exhibited similar defects. Our data indicate that N-glycosylation of N-terminal Asn4 and Asn10 but not of Asn179 in the second extracellular loop is essential for proper folding and, hence, function of FPR. FPR deglycosylation by bacterial glycosidases could be a mechanism by which bacteria compromise host defense. PMID- 12565837 TI - Role of high mobility group protein-1 (HMG1) in amyloid-beta homeostasis. AB - In Alzheimer's disease (AD), fibrillar amyloid-beta (Abeta) peptides form senile plaques associated with activated microglia. Recent studies have indicated that microglial Abeta clearance is facilitated by several activators such as transforming growth factor-beta1 (TGF-beta1). The relationship between microglia and Abeta formation and deposition is still unclear. In the present study, high mobility group protein-1 (HMG1) inhibited the microglial uptake of Abeta (1-42) in the presence and absence of TGF-beta1. In addition, HMG1 bound to Abeta (1-42) and stabilized the oligomerization. In AD brains, protein levels of HMG1 were significantly increased in both the cytosolic and particulate fractions, and HMG1 and Abeta were colocalized in senile plaques associated with microglia. These results suggest that HMG1 may regulate the homeostasis of extracellular Abeta (1 42) and Abeta oligomerization. PMID- 12565838 TI - Mammalian actin binding protein 1 is essential for endocytosis but not lamellipodia formation: functional analysis by RNA interference. AB - Mammalian actin binding protein 1 (mAbp1, also called SH3P7/Hip55) is structurally and functionally related to yeast Abp1 and to cortactin, both of which have been implicated in endocytotic processes. mAbp1 associates through its SH3 domain with dynamin, a large GTPase essential for vesicle fission. To clarify the function of mAbp1, we specifically knocked down its expression in human embryonic kidney 293T cells, using RNA interference (RNAi). Co-transfection of a short interfering RNA (siRNA) together with a plasmid coding for a surface marker, followed by purification of transfected cells, enabled us to obtain a cell population having up to 90% inhibition of mAbp1 expression. In mAbp1-knocked down cells, transferrin (Tf) receptor endocytosis was significantly inhibited and intracellular distribution of the early endosomal compartment was modified. In contrast, in these cells actin and microtubule filaments appeared normal, and formation of lamellipodia induced by active Rac was not inhibited. This study provides definitive evidence that mAbp1 is indispensable for receptor-mediated endocytosis. PMID- 12565839 TI - Archaeal-type rhodopsins in Chlamydomonas: model structure and intracellular localization. AB - Phototaxis in the unicellular green alga Chlamydomonas reinhardtii is mediated by rhodopsin-type photoreceptor(s). Recent expressed sequence tag database from the Kazusa DNA Research Institute has provided the basis for unequivocal identification of two archaeal-type rhodopsins in it. Here we demonstrate that one is located near the eyespot, wherein the photoreceptor(s) has long been thought to be enriched, along with the results of bioinformatic analyses. Secondary structure prediction showed that the second putative transmembrane helices (helix B) of these rhodopsins are rich in glutamate residues, and homology modeling suggested that some additional intra- or intermolecular interactions are necessary for opsin-like folding of the N-terminal ca. 300-aa membrane spanning domains of 712 and 737-aa polypeptides. These results complement physiological and electrophysiological experiments combined with the manipulation of their expression [O.A. Sineshchekov, K.H. Jung, J.H. Spudich, Proc. Natl. Sci. USA 99 (2002) 8689; G. Nagel, D. Olig, M. Fuhrmann, S. Kateriya, A.M. Musti, E. Bamberg, P. Hegemann, Science 296 (2002) 2395]. PMID- 12565840 TI - Human trophoblast noncoding RNA suppresses CIITA promoter III activity in murine B-lymphocytes. AB - Trophoblasts lack expression of all classical MHC antigens. Determination of the mechanism involved could provide insight into selective gene suppression and allograft tolerance. Suppression of class II expression in trophoblasts is secondary to dominant negative trans-acting factors that suppress class II transactivator (CIITA) transcription. We recently described a trophoblast-derived noncoding RNA (TncRNA) that suppresses class II expression. Murine B-cells CH27 were stably transfected with TncRNA and analyzed for MHC class II and CIITA expression by FACS and Northern blots. Functional assessment of CIITA promoter III (pIII) was performed by transient transfection of promoter-reporter constructs. Methylation of pIII was assessed by Southern blots and FACS. TncRNA suppressed constitutive I-Ak and CIITA expression in murine B-cells CH27. The mechanism involves inhibition of CIITA pIII activity. The mechanism does not involve methylation of the promoter. PMID- 12565841 TI - There exist at least 30 human G-protein-coupled receptors with long Ser/Thr-rich N-termini. AB - We report six novel members of the superfamily of human G-protein coupled receptors (GPCRs) found by searches in the human genome databases, termed GPR123, GPR124, GPR125, GPR126, GPR127, and GPR128. Phylogenetic analysis demonstrates that these are additional members of the family of GPCRs with long N-termini, previously termed EGF-7TM, LNB-7TM, B2 or LN-7TM, showing that there exist at least 30 such GPCRs in the human genome. Three of these receptors form their own phylogenetic cluster, while two other places in a cluster with the previously reported HE6 and GPR56 (TM7XN1) and one with EMR1-3. All the novel receptors have a GPS domain in their N-terminus, except GPR123, as well as long Ser/Thr rich regions forming mucin-like stalks. GPR124 and GPR125 have a leucine rich repeat (LRR), an immunoglobulin (Ig) domain, and a hormone-binding domain (HBD). The Ig domain shows similarities to motilin and titin, while the LRR domain shows similarities to LRIG1 and SLIT1-2. GPR127 has one EGF domain while GPR126 and GPR128 do not contain domains that are readily recognized in other proteins beyond the GPS domain. We found several human EST sequences for most of the receptors showing differential expression patterns, which may indicate that some of these receptors participate in central functions while others are more likely to have a role in the immune or reproductive systems. PMID- 12565842 TI - Inhibition of Na+,K+-ATPase by ouabain triggers epithelial cell death independently of inversion of the [Na+]i/[K+]i ratio. AB - Treatment with ouabain led to massive death of principal cells from collecting ducts (C7-MDCK), indicated by cell swelling, loss of mitochondrial function, an irregular pattern of DNA degradation, and insensitivity to pan-caspase inhibitor. Equimolar substitution of extracellular Na(+) by K(+) or choline(+) sharply attenuated the effect of ouabain on intracellular Na(+) and K(+) content but did not protect the cells from death in the presence of ouabain. In contrast to ouabain, inhibition of the Na(+)/K(+) pump in K(+)-free medium increased Na(+)(i) content but did not affect cell survival. In control and K(+)-free medium, ouabain triggered half-maximal cell death at concentrations of approximately 0.5 and 0.05 microM, respectively, which was consistent with elevation of Na(+)/K(+) pump sensitivity to ouabain in K(+)-depleted medium. Our results show for the first time that the death of ouabain-treated renal epithelial cells is independent of the inhibition of Na(+)/K(+) pump-mediated ion fluxes and the [Na(+)](i)]/[K(+)](i) ratio. PMID- 12565843 TI - Kinetic studies of unfolding process of aspergillopepsin II by pH-jump methods. AB - pH-induced unfolding process of the acid proteinase aspergillopepsin II was studied kinetically by stopped-flow methods using circular dichroism and small angle X-ray scattering. Native aspergillopepsin II consists of two polypeptide chains, a light chain and a heavy chain, which are bound noncovalently to each other at acidic pH. Above neutral pH the two chains are known to dissociate and swell due to electrostatic repulsion inside the molecule. The present studies suggested that the polypeptide chains dissociate first and that the dissociated heavy chain expands subsequently. This two-phased pathway is consistent with the accumulation of an intermediate species in the pH-induced unfolding of the enzyme. PMID- 12565844 TI - The footprint of antibody bound to pig cells: evidence of complex surface topology. AB - The disaccharide Gal(alpha)1-3Gal is found on more than 45 different molecules on the endothelium of porcine cells and has recently attracted considerable interest, being the major target recognized by xenoreactive antibodies. In this study, the distribution and topology of Gal(alpha)1-3Gal on porcine endothelial cells was examined to access whether some Gal(alpha)1-3Gal-containing molecules might be preferentially recognized by antibodies binding to Gal(alpha)1-3Gal. Thirteen percent of the Gal(alpha)1-3Gal was found on glycolipid and 87% on glycoproteins. Of all the glycoproteins and glycolipids containing Gal(alpha)1 3Gal, two molecules, fibronectin and the integrin beta1 subunit, were most intensely labeled by galactose oxidase, suggesting that these molecules may be preferentially exposed on the apical surface of the endothelium. Binding of anti Gal(alpha)1-3Gal antibodies to endothelial cell surfaces significantly diminished labeling of fibronectin and the integrin beta1 subunit by galactose oxidase, indicating that these glycoproteins are targets for the antibodies when binding to intact porcine cells. PMID- 12565845 TI - Leucine-764 near the extreme C-terminal end of carnitine palmitoyltransferase I is important for activity. AB - Muscle carnitine palmitoyltransferase I (M-CPTI) catalyzes the conversion of long chain fatty acyl-CoAs to acylcarnitines in the presence of L-carnitine. To determine the role of the C-terminal region of M-CPTI in enzyme activity, we constructed a series of deletion and substitution mutants. The mutants were expressed in the yeast Pichia pastoris, and the effect of the mutations on M-CPTI activity and malonyl-CoA sensitivity was determined in isolated mitochondria prepared from the yeast strains expressing the wild-type and deletion mutants. Deletion of the last 210, 113, 44, 20, 10, and 9 C-terminal amino-acid residues resulted in an inactive M-CPTI, but deletion of the last 8, 7, 6, and 3 C terminal residues had no effect on activity, demonstrating that leucine-764 (L764) is essential for catalysis. Substitution of L764 with alanine caused a 40% loss in catalytic activity, but replacement of L764 with arginine resulted in an 84% loss of activity; substitution of L764 with valine had no effect on catalytic activity. The catalytic efficiency for the L764R mutant decreased by 80% for both substrates. Secondary structure prediction of the M-CPTI sequence identified a 21 amino-acid residue, 744-764, predicted to fold into a coiled-coil alpha-helix in the extreme C-terminal region of M-CPTI that may be important for native folding and activity. In summary, our data demonstrate that deletion of L764 or substitution with arginine inactivates the enzyme, suggesting that L764 may be important for proper folding of M-CPTI and optimal activity. PMID- 12565846 TI - Transcriptional regulation of human Galbeta1,3GalNAc/Galbeta1, 4GlcNAc alpha2,3 sialyltransferase (hST3Gal IV) gene in testis and ovary cell lines. AB - The mRNA expression of sialyltransferase genes is regulated in a cell-type specific manner. The mRNAs of human Galbeta1, 3GalNAc/Galbeta1, 4 GlcNAc alpha2,3 sialyltransferase gene (hST3Gal IV) consist of six isoforms, type A1, A2, B1, B2, B3, and BX. These mRNAs are transcribed from different promoters, pA, pB1, pB2, pB3, and pBX, respectively. Type B mRNAs are expressed in several cells, whereas type A mRNAs are specifically expressed in testis, ovary, and placenta, suggesting that pA promoter activity is especially high in these tissues. We show herein germ-cell specific transcriptional regulation of the hST3Gal IV pA promoter. Using a luciferase assay, pA promoter activity is shown to be high in testis and ovary cell lines. We identified the enhancer region of the pA promoter, located at nt -520 to -420. These results suggest that this element plays a critical role in germ-cell specific regulation of the pA promoter. The results of site-directed mutagenesis suggest that AP2 and c-Ets sites in this region are involved in pA promoter activity, which in turn suggests that the hST3Gal IV gene is regulated in a tissue-restricted fashion at the level of transcription. PMID- 12565847 TI - Amphiphysin1 inhibits vitronectin-mediated cell adhesion, spreading, and migration in vitro. AB - To investigate the regulatory mechanism of cell adhesion, we have searched for cellular inhibitory factors which prevent cell adhesion. The brain cytosol was found to inhibit the adhesion of various transformed cells to the substratum. An inhibitory 120-kDa protein was purified by sequential column chromatography. Peptide sequencing revealed that the protein is identical to amphiphysin1. GST amphiphysin1 suppressed the attachment of HeLa cells to the plate when cells were cultured in the serum-containing medium. Vitronectin, a major cell-adhesive protein in serum and a ligand to alpha(v)beta3 integrin, was responsible for this cell attachment, and the vitronectin action was blocked by GST-amphiphysin1. GST amphiphysin1 also inhibited the vitronectin-mediated spreading and migration of malignant melanoma cells. Furthermore, GST-amphiphysin1 bound directly to vitronectin. These findings point to the interesting possibility that amphiphysin1 could be a useful tool to inhibit cell-adhesive vitronectin. PMID- 12565848 TI - Unique, pH-dependent biphasic band shape of the visible circular dichroism of curcumin-serum albumin complex. AB - Interaction between the plant derived polyphenolic type curcumin molecule having anticarcinogenic, antiinflammatory, and antioxidant activities, and human serum albumin was studied at different pH values by circular dichroism (CD) and electronic absorption spectroscopy. The weak, induced CD spectrum of curcumin-HSA complex measured at pH 7.4 in the visible spectral region shows striking changes upon alkalization; CD spectra collected between pH 7.7 and 9.3 exhibit characteristic, oppositely signed CD band pair according to the visible absorption band of HSA-bound curcumin. At 0.3 curcumin/HSA molar ratio, typical molar CD values are Delta epsilon (496.6nm)+40M(-1)cm(-1) and Delta epsilon (426.8nm)-40M(-1)cm(-1), respectively (pH 9.0, t=37 degrees C). The induced optical activity is attributed to a bent, right-handed chiral conformation of the HSA-bound curcumin molecule within which intramolecular exciton coupling occurs between the electric dipole transition moments of the dissymmetrically juxtaposed feruloyl chromophores. Deprotonation of phenolic OH group(s) of curcumin seems to be the reason leading to the conformational alteration of HSA-bound curcumin. PMID- 12565849 TI - Actomyosin motility on nanostructured surfaces. AB - We have here, for the first time, used nanofabrication techniques to reproduce aspects of the ordered actomyosin arrangement in a muscle cell. The adsorption of functional heavy meromyosin (HMM) to five different resist polymers was first assessed. One group of resists (MRL-6000.1XP and ZEP-520) consistently exhibited high quality motility of actin filaments after incubation with HMM. A second group (PMMA-200, PMMA-950, and MRI-9030) generally gave low quality of motility with only few smoothly moving filaments. Based on these findings electron beam lithography was applied to a bi-layer resist system with PMMA-950 on top of MRL 6000.1XP. Grooves (100-200nm wide) in the PMMA layer were created to expose the MRL-6000.1XP surface for adsorption of HMM and guidance of actin filament motility. This guidance was quite efficient allowing no U-turns of the filaments and approximately 20 times higher density of moving filaments in the grooves than on the surrounding PMMA. PMID- 12565850 TI - Revealing the mode of action of DNA topoisomerase I and its inhibitors by atomic force microscopy. AB - In this study, we used, for the first time, atomic force microscope (AFM) images to investigate the mode of action of DNA topoisomerase I (topo I) in the presence and absence of its inhibitors: camptothecin (CPT) and tyrphostin AG-1387. The results revealed that in the absence of the inhibitors, the enzyme relaxed supercoiled DNA starting from a certain point in the DNA molecules and proceeded in one direction towards one of the edges of the DNA molecule. In addition, the relaxation of the supercoiled DNA is subsequently followed by a knotting event. In the presence of CPT, enzyme-supercoiled DNA complexes in which the enzyme is locked inside a relaxed region of the supercoiled DNA molecule were observed. Tyrphostin AG-1387 altered the DNA relaxation process of topo I producing unique shapes of DNA molecules. AFM images of the topo I protein provided a picture of the enzyme, which resembles its known crystallographic structure. Thus, AFM images provide new information on the mode of action of topo I in the absence and presence of its inhibitors. PMID- 12565851 TI - Down-regulation of Bcl-2-interacting protein BAG-1 confers resistance to anti cancer drugs. AB - BAG-1 was originally identified as a binding partner of anti-apoptotic factor Bcl 2 [Takayama et al., Cell 80 (1995) 279-284]. Exogenous expression of BAG-1 was reported to confer cells resistance to several stresses [Chen et al., Oncogene 21 (2002) 7050]. We have obtained human cervical cancer HeLa cells with down regulated BAG-1 levels by using a highly specific and efficient RNA interference approach. Surprisingly, cells with down-regulated BAG-1 exhibited significantly lower sensitivity against several anti-cancer drugs than parental cells expressing normal levels of the protein. Furthermore, growth rate of the cells was reduced when BAG-1 was down-regulated. Activity of ERK pathway appeared to be decreased in BAG-1 down-regulated cells, as shown by the reduced phosphorylation of ERK1/2 proteins. Taken together resistance against anti-cancer drugs acquired by BAG-1 down-regulated cells may well be accounted for by the retardation of cell cycle progression, implicating the importance of BAG-1 in cell growth regulation. PMID- 12565852 TI - Functional inhibition of the p75 receptor using a small interfering RNA. AB - The neurotrophin receptor p75(NTR) mediates a wide variety of biological effects. Consistent with the function in controlling the survival and neurite formation, p75(NTR) is expressed during the developmental stages of the nervous system. Importantly, p75(NTR) is re-expressed in various pathological conditions and is suggested to contribute to the inhibition of neuronal regeneration and the death of the neurons. Here we develop a tool to knock down the expression of p75(NTR) by employing a small interfering RNA (siRNA). The siRNA for p75(NTR) effectively reduces the expression of endogenous p75(NTR) both in Schwann cells and dorsal root ganglion neurons in vitro. NGF-induced cell death in Schwann cells and the neurite retraction in DRG neurons induced by myelin-associated glycoprotein are attenuated by the siRNA. Inhibition of p75(NTR) in specific pathological conditions by the siRNA may provide a potential therapeutic agent. PMID- 12565853 TI - Adiponutrin mRNA expression in white adipose tissue is rapidly induced by meal feeding a high-sucrose diet. AB - Adiponutrin is a recently identified gene of unknown function that is expressed exclusively in adipose tissues. To provide information about its physiological regulation and possible function, the effect of meal-feeding rats on the expression of adiponutrin mRNA in white adipose tissue was studied. A high sucrose meal increased adiponutrin mRNA levels by at least 5-fold within 3 h. Post-meal levels returned to pre-meal levels with a half-life of about 5 h. The induction was prevented by injection of actinomycin-D prior to the meal. This pattern of expression was very similar to that seen for leptin mRNA. There were only minimal, or no, effects on acrp30/adiponectin, resistin, adipsin, or stearoyl-CoA desaturase 1. Adiponutrin appears more like leptin with respect to its acute regulation by meal-feeding than to any of the other adipokines or to enzymes directly involved in lipogenesis. This suggests that adiponutrin could be involved in overall energy homeostasis, as is leptin. PMID- 12565854 TI - Inhibition of the TEF/TEAD transcription factor activity by nuclear calcium and distinct kinase pathways. AB - Transcription enhancer factor (TEF/TEAD) is a family of four transcription factors that share a common TEA-DNA binding domain and are involved in similar cellular functions, such as cell differentiation and proliferation. All adult tissues express at least one of the four TEAD genes, so this family of transcription factors may be of widespread importance, yet little is known about their regulation. Here we examine the factors that regulate TEAD activity in CHO cells. RT-PCR indicated the presence of TEAD-1, TEAD-3, and both isoforms of TEAD 4, but not TEAD-2. Quantitative measurements showed that TEAD-4 is most abundant, followed by TEAD-3, then TEAD-1. We examined the relative effects of nuclear and cytosolic Ca(2+) on TEAD activity, since TEAD proteins are localized to the nucleus and since free Ca(2+) within the nucleus selectively regulates transcription in some systems. Chelation of nuclear but not cytosolic Ca(2+) increased TEAD activity two times above control. Inhibition of mitogen-activated protein kinase (MAPK) also increased TEAD activity, while cAMP decreased TEAD activity, and protein kinase C had no effect. Together, these results show that nuclear Ca(2+), MAPK, and cAMP each negatively regulate the activity of the TEAD transcription factor. PMID- 12565855 TI - Anti-cachectic effect of ghrelin in nude mice bearing human melanoma cells. AB - Ghrelin is a novel brain-gut peptide that stimulates food intake and body weight gain. We studied the anabolic effect of ghrelin in a cancer cachexia mouse model. SEKI, a human melanoma cell line, was inoculated into nude mice to examine the effects of ghrelin on food intake and body weight. The intraperitoneal administration of ghrelin twice a day (6 nmol/mice/day) for 6 days suppressed weight loss in SEKI-inoculated mice and increased the rate of weight gain in vehicle-treated nude mice. Ghrelin administration also increased food intake in both SEKI- and vehicle-treated mice. Both the weight of white adipose tissue and the plasma leptin concentration were reduced in tumor-inoculated mice compared with vehicle-treated mice; these factors increased following ghrelin administration. The levels of both ghrelin peptide and mRNA in the stomach were upregulated in tumor-inoculated mice. The anabolic effect of ghrelin efficiently reverses the cachexia in mice bearing SEKI human melanoma. Ghrelin therefore may have a therapeutic ability to ameliorate cancer cachexia. PMID- 12565856 TI - Molecular cloning, transcriptional, and expression analysis of the first cellulase gene (cbh2), encoding cellobiohydrolase II, from the moderately thermophilic fungus Talaromyces emersonii and structure prediction of the gene product. AB - A gene (cbh2) encoding cellobiohydrolase II was isolated from the fungus Talaromyces emersonii by rapid amplification of cDNA ends techniques and the equivalent genomic sequence was subsequently cloned. This represents the first report of a key component of the cellulase regulon from this organism. DNA sequencing revealed that cbh2 has an open reading frame of 1377 bp, which encodes a putative polypeptide of 459 amino acids, and is interrupted by seven introns. The deduced amino acid sequence revealed that cbh2 has a modular structure with a predicted molecular mass of 47 kDa and consisting of a fungal type carbohydrate binding module separated from a catalytic domain by a proline/serine/threonine rich linker region. The deduced protein is homologous to fungal cellobiohydrolases in Family 6A of the glycosyl hydrolases. Profiles of cbh2 expression in T. emersonii investigated by Northern blot analysis revealed that expression is regulated at the transcriptional level. Expression of the T. emersonii cbh2 gene is induced by cellulose, xylan, xylose, and gentiobiose and clearly repressed by glucose. Putative regulatory element consensus sequences have been identified in the upstream regulatory sequence of the cbh2 gene including the catabolite repressor element and the activator of cellulase expression (Ace) binding sites. High sequence identity (67%) between the catalytic domain of Cel 6A from Trichoderma reesei and the T. emersonii cbh2 gene product allowed structure prediction for the 3D model of the T. emersonii catalytic domain to be a variant of the classical TIM alpha/beta fold. PMID- 12565857 TI - Self-interaction of heterochromatin protein 1 is required for direct binding to histone methyltransferase, SUV39H1. AB - Heterochromatin protein 1 (HP1) binds to the nucleosome via a methylated lysine residue 9 of histone H3 which is catalyzed by a histone methyltransferase such as SUV39H1. Although co-localization of HP1 and SUV39H1 has been evident in immunostaining and immunoprecipitation experiments, direct protein-protein interactions have remained to be characterized. We examined interactions between mouse HP1 alpha (mHP1 alpha) and SUV39H1 in yeast and in vitro. A yeast two hybrid and a glutathione S-transferase pull-down study indicated that the chromo shadow domain of mHP1 alpha directly interacts with the N-terminal 39 amino acid stretch of SUV39H1. The IY165/168EE mutation in the chromo shadow domain of mHP1 alpha abrogated a self-interaction and this mutant did not interact with SUV39H1. The 13-mer peptide containing a consensus sequence for binding to the dimer surface formed by the chromo shadow domains inhibited interaction between mHP1 alpha and SUV39H1. It seems that self-interaction through the chromo shadow domain of HP1 is crucial for recruitment of SUV39H1 onto nucleosomes. PMID- 12565858 TI - Changes in N-glycosylation of human stromal cells by telomerase expression. AB - It was established that remarkable changes in the N-glycosylation are induced in immortalized cancer cells. Whether changes were induced in human stromal cells immortalized by transfection with the human telomerase catalytic subunit (hTert) cDNA was examined by lectin blot analysis. Morphological appearance and growth rate of the gene-transfected stromal cells were not changed significantly. However, lectin blot analysis of membrane glycoprotein samples showed that bindings of Ricinus communis agglutinin-I (RCA-I) and of leuko-agglutinating phytohemagglutinin to glycoprotein bands increase significantly in the gene transfected cells. No lectin binding was observed when blotted filters were treated with diplococcal beta-1,4-galactosidase or N-glycanase prior to incubation with RCA-I. In contrast, no changes in Coomassie brilliant blue staining and in binding of concanavalin A were obtained between the primary and gene-transfected stromal cells. These results indicate that the highly branched N glycosylation with augmented galactosylation is induced in human stromal cells immortalized by the telomerase expression. PMID- 12565859 TI - cDNA cloning and characterization of Agkistin, a new metalloproteinase from Agkistrodon halys. AB - Agkistin was a new snake venom metalloproteinase (SVMP) gene which was cloned from Agkistrodon halys. Its deduced amino acid sequence has two additional cysteines (Cys407 and Cys426) in the disintegrin domain compared to other RGD containing SVMPs. The full-length gene (Agkistin) and its disintegrin region (named Agkistin-s) were expressed by baculovirus expression system (pFastBac-Htb vector) with His-tag, and their platelet aggregation-inhibition activity was evaluated. The expressed protein Agkistin can also induce apoptosis of HMEC cells in the basal medium after incubated at 37 degrees C for 20 h. PMID- 12565860 TI - Neurotrophic effect of Semaphorin 4D in PC12 cells. AB - Semaphorins provide crucial attractive and repulsive cues involved in axon guidance during neural development. Out of them, Semaphorin 4D (Sema4D) is enriched in the nervous and immune tissues, and acts as proliferative and survival factors of peripheral lymphocytes in the immune system, but is poorly understood in the nervous system. By using PC12 cells which are well known to differentiate into neural cells in response to nerve growth factor (NGF), we found that soluble forms of Sema4D had neurotrophic effects which were inhibited by neutralizing antibodies to Sema4D. Sema4D strikingly potentiated neurite outgrowth in the presence of 50 ng/ml NGF and increased sensitivity to NGF. Cells responded to very low concentrations of NGF in the presence of 1 nM Sema4D. Activation of following signal proteins, protein kinase C (PKC), L-type of voltage-dependent Ca(2+) channel, and phosphatidylinositol (PI) 3-kinase mediated neurotrophic neurite-outgrowth action of Sema4D. These findings suggest a new function of Sema4D as a neurotrophic signal in PC12 cells. PMID- 12565861 TI - Conformational change of the chloroplast ATP synthase on the enzyme activation process detected by the trypsin sensitivity of the gamma subunit. AB - Delta mu H(+) is known to stimulate the enzyme activity of chloroplast ATP synthase in addition to its important role as energy supply for ATP synthesis. In the present study, we focused on the relationship between the proton translocation via the membrane sector of ATP synthase, F(o), and the conformational change of the central stalk subunit gamma. The conformational change of CF(1) mainly at the gamma subunit was induced by the proton flow via F(o) in the absence of substrates. The effects of inhibitors on CF(o) or CF(1) for this conformational change were also examined. The observed conformational change was partially suppressed by ADP binding. From these results, we propose the Delta mu H(+)-dependent conformational change of CF(1) on the enzyme activation process, which is affected by both ADP binding to the catalytic sites and proton flow via F(o) portion. PMID- 12565862 TI - EXT gene family member rib-2 is essential for embryonic development and heparan sulfate biosynthesis in Caenorhabditis elegans. AB - EXT gene family members including EXT1, EXT2, and EXTL2 are glycosyltransferases required for heparan sulfate biosynthesis. To examine the biological functions of rib-2, a member of the Caenorhabditis elegans EXT gene family, we generated a mutant worm lacking the rib-2 gene using the UV-TMP method followed by sib selection. Inactivation of rib-2 alleles induced developmental abnormalities in F2 and F3 homozygous worms, while F1 heterozygotes showed a normal morphology. The F2 homozygous progeny generated from the F1 heterozygous hermaphrodites somehow developed to adult stage but exhibited abnormal characteristics such as developmental delay and egg-laying defects. The F3 homozygous progeny from the F2 homozygous hermaphrodites showed early developmental defects and most of the F3 worms stopped developing during the gastrulation stage. Whole-mount staining analysis for heparan sulfate using Toluidine blue (pH 2.5) revealed a defect of heparan sulfate biosynthesis in the F2 homozygotes. The analysis using fluorometric post-column high-performance liquid chromatography also uncovered reduced production of heparan sulfate in the rib-2 mutant. These results indicate that rib-2 is essential for embryonic development and heparan sulfate biosynthesis in C. elegans. PMID- 12565863 TI - Effect of cisplatin treatment on speckled distribution of a serine/arginine-rich nuclear protein CROP/Luc7A. AB - The C-half of cisplatin resistance-associated overexpressed protein (CROP), an SR related protein, comprises domains rich in arginine and glutamate residues (RE domain), and is rich in arginine and serine residues (RS domain). We analyzed the role of the individual domains of CROP in cellular localization, subnuclear localization, and protein-protein interaction. CROP fused with green fluorescent protein, GFP-CROP, localized exclusively to the nucleus and showed a speckled intranuclear distribution. The yeast two-hybrid system revealed that CROP interacted with SF2/ASF, an SR protein involved in RNA splicing, as well as CROP itself. The RE and RS domains were necessary for both the intranuclear speckled distribution and the protein-protein interaction. CROP was phosphorylated by mSRPK1, mSRPK2, and Clk1 in vitro, and when cells were treated with cisplatin the subnuclear distribution of GFP-CROP was changed. These results suggest that cisplatin affects RNA splicing by changing the subnuclear distribution of SR proteins including CROP. PMID- 12565864 TI - Induction of hepatic injury by hepatitis C virus-specific CD8+ murine cytotoxic T lymphocytes in transgenic mice expressing the viral structural genes. AB - In the present study, we generated killer cells specific for hepatitis C virus (HCV) structural protein by re-stimulation of immune spleen cells from H-2(d) haplotype transgenic (Tg) mice, expressing the core, E1, E2, and NS2 genes of HCV regulated by the Cre/loxP switching system. The generated killer cells were conventional CD8(+)L(d) class-I MHC molecule-restricted cytotoxic T lymphocytes (CTLs) and specific for the HCV E1 structural protein. Because the CTLs could also kill hepatocytes from the Tg mice expressing HCV structural proteins in vitro, we attempted to transfer those CTLs intravenously into interferon regulatory factor-1 (IRF-1) negative, CD8-deficient Tg mice representing the HCV structural genes on hepatocytes to examine whether the inoculated CD8(+) CTLs can eliminate hepatocytes expressing the HCV genes in vivo. We observed an elevation of serum ALT level as well as damage of the liver tissue histologically. To our knowledge, this is the first demonstration to show that HCV-specific CD8(+) CTLs specifically attack hepatocytes expressing the HCV structural proteins both in vitro and in vivo. PMID- 12565865 TI - Chondrogenesis enhanced by overexpression of sox9 gene in mouse bone marrow derived mesenchymal stem cells. AB - We investigated chondrogenesis of cell-mediated sox9 gene therapy as a new treatment regimen for cartilage regeneration. pIRES2-EGFP vector containing a full-length mouse sox9 cDNA was transfected into bone marrow-derived mesenchymal stem cells (MSCs) by lipofection and chondrogenic differentiation of these cells was evaluated. In vitro high density micromass culture of these sox9 transfected MSCs demonstrated that a matrix-rich micromass aggregate with EGFP expressing MSCs was positively stained by Alcian blue and type II collagen. Next, sox9 transfected MSCs were loaded into the diffusion chamber and transplanted into athymic mice to analyze in vivo chondrogenesis. A massive tissue formation in about 2mm diameter was visible in the chamber after 4 weeks transplantation. Histological examinations demonstrated that both Alcian blue and type II collagen were positively stained in the extracellular matrix of the mass while type X collagen was not stained. These results indicated that cell-mediated sox9 gene therapy could be a novel strategy for hyaline cartilage damage. PMID- 12565866 TI - PACAP and NGF cooperatively enhance choline acetyltransferase activity in postnatal basal forebrain neurons by complementary induction of its different mRNA species. AB - Both nerve growth factor (NGF) and pituitary adenylate cyclase activating polypeptide (PACAP) have neurotrophic effects on basal forebrain cholinergic neurons. They promote differentiation, maturation, and survival of these cholinergic neurons in vivo and in vitro. Here we report on the cooperative effects of NGF and PACAP on postnatal, but not embryonic, cholinergic neurons cultured from rat basal forebrain. Combined treatment with NGF, brain-derived neurotrophic factor (BDNF), neurotrophin-4 (NT-4), and PACAP induced an additive increase in choline acetyltransferase (ChAT) activity. There were no cooperative effects on the number of cholinergic neurons, suggesting that ChAT mRNA expression had been induced in each cholinergic neuron. Further analysis revealed that NGF and PACAP led to complementary induction of different ChAT mRNA species, thus enhancing total ChAT mRNA expression. These results explain the cooperative neurotrophic action of NGF and PACAP on postnatal cholinergic neurons. PMID- 12565867 TI - Identification of three novel salicylate 1-hydroxylases involved in the phenanthrene degradation of Sphingobium sp. strain P2. AB - Five sets of large and small subunits of terminal oxygenase (ahdA1[a-e] and ahdA2[a-e]) and a single gene set encoding ferredoxin (ahdA3) and ferredoxin reductase (ahdA4) were found to be scattered through 15.8- and 14-kb DNA fragments of phenanthrene-degrading Sphingobium sp. strain P2. RT-PCR analysis indicated the inducible and specific expression of ahdA3, ahdA4, and three sets of genes for terminal oxygenase (ahdA1[c-e] and ahdA2[c-e]) in this strain grown on phenanthrene. The biotransformation experiments with resting cells of Escherichia coli JM109 harboring recombinant ahd genes revealed that AhdA2cA1c, AhdA1dA2d, and AhdA1eA2e can all function as a salicylate 1-hydroxylase which converts salicylate, a metabolic intermediate of phenanthrene, to catechol in cooperation with the electron transport proteins AhdA3A4. The first two oxygenases exhibited a broad range of substrate specificities such that they also catalyzed the hydroxylation of methyl- and chloro-substituted salicylates to produce their corresponding substituted catechols. PMID- 12565868 TI - Thymidine phosphorylase inhibits apoptosis induced by cisplatin. AB - An angiogenic factor, platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP), stimulates the chemotaxis of endothelial cells and confers resistance to apoptosis induced by hypoxia. 2-Deoxy-D-ribose, a degradation product of thymidine generated by TP, partially prevents hypoxia induced apoptosis. TP is expressed at higher levels in tumor tissues compared to the adjacent non-neoplastic tissues in a variety of human carcinomas. High expression of TP is associated with an unfavorable prognosis. To investigate the effect of TP on cisplatin-induced apoptosis, human leukemia Jurkat cells were transfected with wild-type or mutant (L148R) TP cDNA. TP inhibited a number of steps in the cisplatin-induced apoptotic pathway, activation of caspases 3 and 9 and mitochondrial cytochrome c release. These findings suggest a mechanism by which TP confers resistance to apoptosis induced by cisplatin. Moreover, mutant TP that has no enzymatic activity also suppressed cisplatin-induced apoptosis. These findings indicate that TP has cytoprotective functions against cytotoxic agents which are independent of its enzymatic activity. PMID- 12565869 TI - Isolation of a large thaumatin-like antifungal protein from seeds of the Kweilin chestnut Castanopsis chinensis. AB - A protein with an N-terminal sequence showing a much lesser extent of homology than French bean and kiwi fruit thaumatin-like proteins (TLPs) to other TLPs, and possessing a molecular mass of 30 kDa which is considerably higher than those of previously reported TLPs, has been purified from the seeds of the chestnut Castanopsis chinensis Hance. The protein was unadsorbed on DEAE-cellulose in 10 mM Tris-HCl buffer (pH 7.3), and adsorbed on Affi-gel blue gel in the same buffer, on CM-cellulose in 10 mM ammonium acetate buffer (pH 4.5), and on Mono S in 20 mM ammonium acetate buffer (pH 5.5). A highly purified protein preparation was obtained after fractionation on the first three chromatographic media. Castanopsis TLP appeared as a single band (30 kDa) in sodium dodecyl sulfate polyacrylamide gel electrophoresis and as a single peak (30 kDa) in gel filtration on Superdex 75 by fast protein liquid chromatography. The TLP exerted antifungal activity against Botrytis cinerea, Fusarium oxysporum, Mycosphaerella arachidicola, and Physalospora piricola, with an IC(50) of 0.5 microM against F. oxysporum. Castanopsis TLP was more potent than French bean and kiwi fruit TLPs in its antifungal activity toward F. oxysporum and M. arachidicola. The antifungal activity of Castanopsis TLP remained essentially unaltered after incubation at 40 degrees C for 10 min, was reduced after incubation at 60 degrees C, and disappeared after treatment at 80 degrees C. The antifungal activity underwent a decline after treatment with trypsin (enzyme:substrate ratio 1:100) at 37 degrees C for 1h but some activity remained. Castanopsis TLP exhibited a much more potent inhibitory activity on HIV-1 reverse transcriptase (IC(50) = 1.6 microM) than kiwi fruit TLP (IC(50) > or = 27 microM). Castanopsis TLP was obtained with a yield of 20 mg from 1 kg chestnut seeds. PMID- 12565870 TI - VEGFR-2-specific ligand VEGF-E induces non-edematous hyper-vascularization in mice. AB - VEGF family members play important roles in angiogenesis and vascular permeability. VEGF-A-transgenic mice showed an increased vascularization with edema due to hyper-vascular permeability and subcutaneous hemorrhage as side effects. VEGF-A binds and activates two receptors, VEGFR-1 (Flt-1) and VEGFR-2 (KDR/Flk-1). To dissect the signals of these two receptors, we generated transgenic mice overexpressing either the VEGFR-2-specific ligand VEGF-E(NZ-7) or VEGFR-1-specific ligand PlGF-II under the control of the Keratin-14 promoter. VEGF-E-mice showed a significant increase in vascularization (about 10-fold compared to control mice) in subcutaneous tissues, whereas PlGF-mice showed only a 2-3-fold increase. Interestingly, VEGF-E-mice did not show any clear edematous lesions or hemorrhagic spots on the skin. Microscopically, VEGF-E-induced capillary networks have a well organized structure with the recruitment of pericytes. These results indicate that VEGF-E is a new angiogenic agent with less side effects for clinical usage. PMID- 12565871 TI - PCIF1, a novel human WW domain-containing protein, interacts with the phosphorylated RNA polymerase II. AB - Phosphorylation of the carboxy-terminal domain (CTD) of RNA polymerase II (RNAP II) largest subunit has an important role in transcription elongation and in coupling transcription to pre-mRNA processing. To identify proteins that can directly bind to the phosphorylated CTD, we screened a human cDNA expression library using 32P-labeled CTD as a probe. Here we report the cloning and characterization of a novel human WW domain-containing protein, PCIF1 (phosphorylated CTD interacting factor 1). PCIF1 is composed of 704 amino acids. The WW domain of PCIF1 can directly and preferentially bind to the phosphorylated CTD compared to the unphosphorylated CTD. PCIF1 binds to the hyperphosphorylated RNAP II (RNAP IIO) in vitro and in vivo. Double immunofluorescence labeling in HeLa cells demonstrated that PCIF1 and endogenous RNAP IIO are co-localized in the cell nucleus. Thus, PCIF1 may play a role in mRNA synthesis by modulating RNAP IIO activity. PMID- 12565872 TI - Cloning and characterization of a functional promoter of the human SOCS-3 gene. AB - SOCS-3 is a member of a newly discovered protein family that inhibits LIF activated Janus kinase (JAK)-signal transducers and activators of transcription (STAT) signaling in a negative auto-regulatory manner. In this study, we have cloned and characterized the promoter region of the human SOCS-3 gene. This region is approximately 1.1 kbp in length and consists of two putative STAT binding elements, a G-rich element, and a putative TATA box. These elements are highly conserved in both murine and rat SOCS-3 promoters. Functional analysis of this region shows that the whole fragment (approximately 1.1 kbp) has high basal promoter activity and is responsive to growth factors. We also found that the wild type SOCS-3 promoter construct has significantly greater activity in non small-cell lung cancer cell lines than in normal cells in accordance with STAT3 disregulation in these cells. Cloning of the human SOCS-3 promoter should help uncover mechanisms of regulation of the JAK-STAT pathway in human cancer. PMID- 12565873 TI - Nedd8-modification of Cul1 is promoted by Roc1 as a Nedd8-E3 ligase and regulates its stability. AB - SCF is a ubiquitin ligase and is composed of Skp1, Cul1, F-box protein, and Roc1. The catalytic site of the SCF is the Cul1/Roc1 complex and RING-finger protein Roc1. It was shown earlier that when Cul1 was co-expressed with Roc1 in Sf-9 cells in a baculovirus protein expression system, Cul1 was highly neddylated in the cell, suggesting that Roc1 may function as a Nedd8-E3 ligase. However, there is no direct evidence that Roc1 is a Nedd8-E3 in an in vitro enzyme system. Here we have shown that Roc1 binds to Ubc12, E2 for Nedd8, but not to Ubc9, E2 for SUMO-1 and Roc1 RING-finger mutant, H77A, did not bind to Ubc12. In in vitro neddylation system using purified Cul1/Roc1 complex expressed in bacteria, Roc1 promotes neddylation of Cul1. These results demonstrate that Roc1 functions as a Nedd8-E3 ligase toward Cul1. Furthermore, Roc1 and Cul1 were ubiquitinylated in a manner dependent on the neddylation of Cul1 in vitro. In addition, Cul1 was degraded through the ubiquitin-proteasome pathway, and a non-neddylated mutant Cul1, K720R, was more stable than wild-type in intact cells. Thus, neddylation of Cul1 might regulate SCF function negatively via degradation of Cul1/Roc1 complex. PMID- 12565874 TI - Novel selective melanocortin 4 receptor antagonist induces food intake after peripheral administration. AB - We synthesized a new series of small cyclic melanocyte-stimulating hormone (MSH) analogues and screened them for binding affinity at the four MSH binding melanocortin (MC) receptors. We identified a novel substance HS131, with about 20 fold higher affinity for the MC4 receptor than the MC3 receptor. This substance proved to be antagonist for all the four MC receptors in a cAMP assay. HS131 is a six amino acid long peptide, has a molecular weight below 1000, and has only two amino acids in common with the natural MSH peptides. HS131 potently and dose dependently increased food intake after i.c.v. administration. Moreover, s.c. administration of HS131 (1.0 mg/kg) increased food intake, suggesting that HS131 may be able to pass the blood brain barrier. This cyclic low molecular weight peptidomimetic will enable studies of the functional role of the MC4 receptors by peripheral administration and it may be used as a template for further development of low molecular weight substances for the MC receptors. PMID- 12565875 TI - A human cell surface receptor activated by free fatty acids and thiazolidinedione drugs. AB - Fatty acids, which are essential nutritional components, are also involved in cardiovascular and metabolic diseases. Here we report a human cell surface receptor that we name free fatty acid receptor (FFAR), because it is specifically activated by medium to long-chain free fatty acids. The receptor belongs to the class of seven-transmembrane, G-protein coupled receptors (GPCRs) and also mediates responses to antidiabetic drugs of the thiazolidinedione type. It is expressed in skeletal muscle, heart, liver, and pancreatic beta-cells. Stimulation of FFAR increases the intracellular calcium concentration in cells expressing the receptor in a native (pancreatic beta-cell line) or in a recombinant form. In view of the nature of the activating substances, their physiological role in the body, and the tissue distribution of FFAR we suggest the term "nutrient sensing receptor" for receptors acting at the interface between dietary components and signalling molecules. PMID- 12565876 TI - Methylene blue photosensitized oxidation of hypotaurine in the presence of azide generates reactive nitrogen species: formation of nitrotyrosine. AB - In our previous study on the hypotaurine (HTAU) oxidation by methylene blue (MB) photochemically generated singlet oxygen (1O2) we found that azide, usually used as 1O2 quencher, produced, instead, an evident enhancing effect on the oxidation rate [L. Pecci, M. Costa, G. Montefoschi, A. Antonucci, D. Cavallini, Biochem. Biophys. Res. Commun. 254 (1999) 661-665]. We show here that this effect is strongly dependent on pH, with a maximum at approximately pH 5.7. When the MB photochemical system containing HTAU and azide was performed in the presence of tyrosine, 3-nitrotyrosine was produced with maximum yield at pH 5.7, suggesting that azide, by the combined action of HTAU and singlet oxygen, generates nitrogen species which contribute to tyrosine nitration. In addition to HTAU, cysteine sulfinic acid, and sulfite were found to induce the formation of 3-nitrotyrosine. No detectable tyrosine nitration was observed using taurine, the oxidation product of HTAU, or thiol compounds such as cysteine and glutathione. It is shown that during the MB photooxidation of HTAU in the presence of azide, nitrite, and nitrate are produced. Evidences are presented, indicating that nitrite represents the nitrogen species involved in the production of 3-nitrotyrosine. A possible mechanism accounting for the enhancing effect of azide on the photochemical oxidation of HTAU and the production of nitrogen species is proposed. PMID- 12565877 TI - Inhibition of amino acid-mTOR signaling by a leucine derivative induces G1 arrest in Jurkat cells. AB - We have previously demonstrated that N-acetylleucine amide, a derivative of L leucine, inhibits leucine-induced p70(S6k) activation in a rat hepatoma cell line. In the present study, we investigated whether N-acetylleucine amide is capable of inhibiting amino acid-mTOR signaling. N-Acetylleucine amide caused cell cycle arrest at G1 stage in Jurkat cells, a human leukemia T cell line, concomitant with the inhibition of serum-induced p70(S6k) activation and p27 degradation. Treatment of Jurkat cells with this compound also exhibited dephosphorylation of retinoblastoma protein. These effects are similar to the inhibitory effects of rapamycin on amino acid-mTOR signaling pathway and suggest that N-acetylleucine amide acts as a rapamycin-like reagent to inhibit cell cycle progression in Jurkat cells. PMID- 12565878 TI - Upregulation of cAMP is a new functional signal pathway of Klotho in endothelial cells. AB - We measured angiotensin I-converting enzyme (ACE) activity in a human endothelial cell to characterize the intracellular signal pathways of Klotho. COS-1 cells transfected with naked mouse membrane-form klotho plasmid DNA (pCAGGS-klotho) translated proper Klotho protein. This translated Klotho protein was secreted into the culture medium. Furthermore, ACE activity in human umbilical vein endothelial cells (HUVEC) was upregulated when HUVEC were co-cultured with COS-1 cells that were pre-transfected with pCAGGS-klotho. The conditioned medium from COS-1 cells pre-transfected with pCAGGS-klotho also dose-dependently upregulated ACE in HUVEC. In addition, the conditioned medium induced time- and dose dependent enhancement of cAMP production in HUVEC. Rp-cAMP, an inhibitor of cAMP dependent protein kinase A (PKA), inhibited the upregulation of ACE by Klotho protein. Our results suggest that mouse membrane-form Klotho protein acts as a humoral factor to increase ACE activity in HUVEC via a cAMP-PKA-dependent pathway. These findings may provide a new insight into the mechanism of Klotho protein. PMID- 12565879 TI - Characterization of the Escherichia coli YedU protein as a molecular chaperone. AB - We have cloned, purified to homogeneity, and characterized as a molecular chaperone the Escherichia coli YedU protein. The purified protein shows a single band at 31 kDa on SDS-polyacrylamide gels and forms dimers in solution. Like other chaperones, YedU interacts with unfolded and denatured proteins. It promotes the functional folding of citrate synthase and alpha-glucosidase after urea denaturation and prevents the aggregation of citrate synthase under heat shock conditions. YedU forms complexes with the permanently unfolded protein, reduced carboxymethyl alpha-lactalbumin. In contrast to DnaK/Hsp70, ATP does not stimulate YedU-dependent citrate synthase renaturation and does not affect the interaction between YedU and unfolded proteins, and YedU does not display any peptide-stimulated ATPase activity. We conclude that YedU is a novel chaperone which functions independently of an ATP/ADP cycle. PMID- 12565880 TI - Decreased mature adrenomedullin levels in feto-maternal tissues of pregnant women with histologic chorioamnionitis. AB - Adrenomedullin (AM) gains its bioactivity by amidation at its C-terminal, forming "mature AM." The mature AM and the expression of AM receptor component mRNAs, receptor activity-modifying protein 2 and calcitonin receptor-like receptor, from feto-maternal tissues of normal pregnant women and women with histologic chorioamnionitis were examined to clarify the pathophysiological features of this intrauterine infection. Samples of the placenta and fetal membranes were obtained from 10 normal pregnant women and eight women with histologic chorioamnionitis under informed consent. Mature AM in the fetal membranes was significantly lower in patients with chorioamnionitis than in normal pregnant women. On the other hand, there were no differences in mature AM levels in the placenta between the two groups. The total AM levels as a sum of mature and immature AM were not significantly different between the two groups in either area. The ratio of mature AM/total AM was significantly decreased in the fetal membranes of the patients with chorioamnionitis compared with normal pregnancies, but not in the placenta. Also, levels of mature AM were negatively correlated with C-reactive protein concentrations. The present results thus suggested that mature AM may have some role in chorioamnionitis. PMID- 12565881 TI - Molecular cloning, structure, and testis-specific expression of MFSJ1, a member of the DNAJ protein family, in the Japanese monkey (Macaca fuscata). AB - A strong signal of cDNA product was identified in adult and senile testes of the Japanese monkeys (Macaca fuscata) using differential display PCR analysis. Its full-length cDNA was molecular-cloned by RT-PCR using adult testis mRNA as templates. The predicted open reading frame encoded a protein of 242 amino-acid residues. It contained J domain in the NH(2) terminal region and Gly/Phe-rich domain in the middle of protein, which are typical structural domains of the DnaJ protein family. We named this gene, MFSJ1, for spermatogenic cell-specific DNAJ homolog in the Japanese monkey. Northern blot analysis of RNAs from various somatic and germinal tissues revealed that the MFSJ1 gene is specifically expressed in testis and is active at adult and senile stages but is scarcely expressed at the juvenile stage. In situ hybridization revealed that the MFSJ1 gene is expressed mainly in spermatids and the expressional potential is maintained from adult to senile stages. MFSJ1 was found to have high similarity (71% identity) with MSJ1, mouse spermatogenic cell-specific DnaJ homolog. Although this type of DnaJ-like protein has not been found in other mammals, it may be essential for mammalian spermatogenesis. PMID- 12565882 TI - Rescue of molybdenum cofactor biosynthesis in gephyrin-deficient mice by a Cnx1 transgene. AB - Gephyrin is a bifunctional protein which is essential for both synaptic clustering of inhibitory neurotransmitter receptors in the central nervous system and the biosynthesis of the molybdenum cofactor (MoCo) in peripheral tissues. Mice deficient in gephyrin die early postnatally and display a loss of glycine receptors (GlyRs) and many GABA(A) receptor (GABA(A)R) subtypes from postsynaptic sites. In addition, the activities of the MoCo-dependent enzymes xanthine dehydrogenase and sulfite oxidase are reduced to background levels in the liver and intestine of these animals. To genetically separate the different consequences of gephyrin deficiency, we expressed a transgene of the plant homolog Cnx1, known to rescue mammalian MoCo deficiency, on the background of gephyrin knockout mice. Cnx1 partially restored sulfite oxidase activity in the liver of the transgenic animals, whereas early lethality and the loss of GlyR clustering were unaltered. Our data suggest that the loss of neurotransmitter receptor clustering at inhibitory synapses causes the early lethality of gephyrin deficient mice. PMID- 12565883 TI - AF2 interaction with Ascaris suum body wall muscle membranes involves G-protein activation. AB - KHEYLRF-NH(2) (AF2) is the most abundant FMRFamide-related peptide (FaRP) in Ascaris suum and also in many other parasitic and free-living nematodes. The AF2 abundance in the highly diverse nematodes and its potent and profound effects on the neuromuscular systems make AF2 and its receptor(s) very attractive targets for the discovery of novel broad-spectrum anthelmintics. Although FaRP receptors are believed to belong to the large family of G-protein coupled receptors (GPCRs), to date no AF2 receptor(s) have been cloned so there is no final proof to show that they are indeed G-protein coupled. In this study, using A. suum body wall muscle membranes, we showed that: (1) AF2 effectively (EC(50) 57 nM) induced a dose-dependent stimulation of [35S]GTP gamma S binding to the membranes, which is a hallmark of G-protein activation; (2) the high affinity binding of [125I Tyr(4)]AF2 was inhibited in a dose-dependent manner by GTP with a K(i) of 10.5 nM (so-called guanine nucleotide effect, characteristic for GPCRs). Collectively, our results provide direct evidence for G-protein involvement in AF2-triggered receptor activation and thus confirm that the receptor for AF2 in A. suum is a GPCR. PMID- 12565884 TI - Novel calcitonin-(8-32)-sensitive adrenomedullin receptors derived from co expression of calcitonin receptor with receptor activity-modifying proteins. AB - We tested whether heterodimers comprised of calcitonin (CT) receptor lacking the 16-amino acid insert in intracellular domain 1 (CTR(I1-)) and receptor activity modifying protein (RAMP) can function not only as calcitonin gene-related peptide (CGRP) receptors but also as adrenomedullin (AM) receptors. Whether transfected alone or together with RAMP, human (h)CTR(I1-) appeared mainly at the surface of HEK-293 cells. Expression of CTR(I1-) alone led to significant increases in cAMP in response to hCGRP or hAM, though both peptides remained about 100-fold less potent than hCT. However, the apparent potency of AM, like that of CGRP, approached that of CT when CTR(I1-) was co-expressed with RAMP. CGRP- or AM evoked cAMP production was strongly inhibited by salmon CT-(8-32), a selective amylin receptor antagonist, but not by hCGRP-(8-37) or hAM-(22-52), antagonists of CGRP and AM receptors, respectively. Moreover, the inhibitory effects of CT-(8 32) were much stronger in cells co-expressing CTR(I1-) and RAMP than in cells expressing CTR(I1-) alone. Co-expression of CTR(I1-) with RAMP thus appears to produce functional CT-(8-32)-sensitive AM receptors. PMID- 12565885 TI - Stable monomeric intermediate with exposed Cys-119 is formed during heat denaturation of beta-lactoglobulin. AB - The role of the free sulfhydryl group of beta-lactoglobulin in the formation of a stable non-native monomer during heat-treatment of beta-lactoglobulin solutions was investigated. Two concomitant events occurred at the earlier stage of heating: unfolding of native globular monomer and intramolecular sulfhydryl/disulfide exchange reaction. Thus, two denatured monomeric species were formed: a non-native monomer with exposed Cys-121 (Mcys121) which became reversible after cooling, and a stable non-native monomer with exposed Cys-119 (Mcys119) which exhibited both a larger hydrodynamic conformation than native monomer and low solubility at pH 4.7. The results also show that the formation of these monomeric species throughout heat-induced denaturation of native beta-lg monomers is faster than their subsequent aggregation. A mechanism describing the behavior of beta-lg denaturation/aggregation during heat-treatment under selected conditions (5.8 mg/ml, low ionic strength, pH 6.6, 85 degrees C) is presented. PMID- 12565886 TI - An intronic alternative promoter of the human lactoferrin gene is activated by Ets. AB - Lactoferrin expresses in a variety of tissues and involves in various aspects of host defense mechanisms. The lactoferrin gene is differentially regulated through multiple signaling pathways. Recently, an alternative form of human lactoferrin mRNA (Delta LF) was found in normal human tissues but absent from the tumor cells. In this study, we identified the transcription start sites of the Delta LF in mammary gland and bone marrow and demonstrated that the Delta LF is the product of an alternative (P2) promoter present in the first intron of the lactoferrin gene. The P2 promoter has high activity in Jurkat and U937 and low activity in RL95-2 and HEC-1B cell lines. Nonetheless, the promoter activity in HEC-1B cells was dramatically enhanced with overexpression of the Ets-1 transcription factor. The GFP-tagged lactoferrin is present in the cytoplasm whereas GFP-tagged Delta LF is found in both nucleus and cytoplasm as examined by fluorescence and confocal microscopy. PMID- 12565887 TI - Flavonoid glycoside: a new inhibitor of eukaryotic DNA polymerase alpha and a new carrier for inhibitor-affinity chromatography. AB - Two flavonoid glycosides, kaempferol 3-O-(6"-acetyl)-beta-glucopyranoside (KAG) and quercetin 3-O-(6"-acetyl)-beta-glucopyranoside (QAG), were found to be inhibitors of eukaryotic DNA polymerases from a Japanese vegetable, Petasites japonicus. These compounds inhibited the activities of mammalian replicative DNA polymerases (i.e., pol alpha, delta, and epsilon), but not other pol beta, eta, kappa, and lambda activities. KAG was a stronger inhibitor and more selective to pol alpha than QAG. The IC(50) values of KAG for pol alpha, delta, and epsilon were 41, 164, and 127 microM, respectively. The pol alpha inhibition by KAG was non-competitive with respect to both the DNA template-primer and the dNTP substrate. KAG and QAG did not influence the activities of prokaryotic DNA polymerases or other mammalian DNA metabolic enzymes such as human immunodeficiency virus type 1 reverse transcriptase, human telomerase, human DNA topoisomerase I and II, T7 RNA polymerase, and bovine deoxyribonuclease I. Therefore, we concluded that these flavonoid glycosides are moderate replicative DNA polymerase inhibitors leaning more relatively to pol alpha, and could be used as chromatographic carriers to purify the DNA polymerases rather than cytotoxic agents. We then made a KAG-conjugated column such as the epoxy-activated Sepharose 6B. In the column, pol alpha was selectively adsorbed and eluted. PMID- 12565888 TI - Mutational analysis of the Menkes copper P-type ATPase (ATP7A). AB - The Menkes protein (ATP7A; MNK) is a ubiquitous human copper-translocating P-type ATPase and it has a key role in regulating copper homeostasis. Previously we characterised fundamental steps in the catalytic cycle of the Menkes protein. In this study we analysed the role of several conserved regions of the Menkes protein, particularly within the putative cytosolic ATP-binding domain. The results of catalytic studies have indicated an important role of 1086His in catalysis. Our findings provide a biochemical explanation for the most common Wilson disease-causing mutation (H1069Q in the homologous Wilson copper translocating P-type ATPase). Furthermore, we have identified a unique role of 1230Asp, within the DxxK motif, in coupling ATP binding and acylphosphorylation with copper translocation. Finally, we found that the Menkes protein mutants with significantly reduced catalytic activity can still undergo copper-regulated exocytosis, suggesting that only the complete loss of catalytic activity prevents copper-regulated trafficking of the Menkes protein. PMID- 12565889 TI - Essential roles for NF-kappa B and a Toll/IL-1 receptor domain-specific signal(s) in the induction of I kappa B-zeta. AB - I kappa B-zeta, a new negative-regulator of nuclear factor-kappa B (NF-kappa B), is strongly induced by lipopolysaccharide or interleukin-1 beta stimulation, but not by tumor necrosis factor-alpha. Here, we analyzed the mechanisms for transcriptional induction of I kappa B-zeta. I kappa B-zeta mRNA was induced by overexpression of MyD88 or TRAF6, but not TRAF2. Stimulation of macrophages with peptidoglycan or CpG DNA, which activated Toll-like receptor 2 or 9, respectively, also resulted in I kappa B-zeta induction. Thus, activation of the MyD88-dependent signaling pathway, commonly found downstream of different Toll/interleukin-1 receptor (TIR) domains, is sufficient for I kappa B-zeta induction. The induction was inhibited by treatment with various inhibitors of NF kappa B activation or by overexpressing I kappa B-alpha or beta, indicating essential roles for NF-kappa B in I kappa B-zeta induction. However, overexpression of the NF-kappa B subunits induced I kappa B-alpha, but not I kappa B-zeta. These results indicate the existence of another signal essential for I kappa B-zeta induction, which is specifically mediated by the TIR domain mediated signaling pathway. PMID- 12565890 TI - Centaurin-alpha 1 associates in vitro and in vivo with nucleolin. AB - Centaurin-alpha(1) was originally described as a binding partner for phosphoinositides. In spite of the presence of a putative ADP-ribosylation factor (ARF) GTPase-activating protein (GAP) domain, no ARF-GAP activity has been attributed to centaurin-alpha(1) so far. Thus the function of this protein remains to be determined. In order to better understand its intracellular role, we aimed to identify centaurin-alpha(1) partners. Using affinity chromatography followed by mass spectrometry analysis, we identified several potential centaurin alpha(1) protein partners. Nucleolin, a nucleolar protein involved in ribosome biosynthesis, was the main centaurin-alpha(1) interacting protein. The interaction between centaurin-alpha(1) and nucleolin was confirmed by Western blot analysis and GST pull down assays. Moreover, we have shown that ectopically expressed centaurin-alpha(1) associates in vivo with endogenous nucleolin in human embryonic kidney 293 cells. In addition, the association between nucleolin and centaurin-alpha(1) was disrupted by RNAse treatment, indicating that RNA integrity was necessary for their binding. This suggested that centaurin-alpha(1) was part of a ribonucleoprotein complex. PMID- 12565891 TI - Identification of new human coding steroid receptor RNA activator isoforms. AB - SRA is a steroid receptor co-activator which acts as a functional RNA and is classified as belonging to the growing family of functional non-coding RNAs. None of the different SRA transcripts described to date encode a detectable SRA protein following in vitro and in vivo translation experiments. We have identified three new SRA-RNA isoforms differing mainly from the originally cloned SRA by an extended 5(') extremity. These long SRA isoforms, able to encode a stable protein in vitro, led to the production in vivo of a nuclear protein when transfected into the MCF-7 human breast cancer cell line. Reverse-transcription polymerase chain reaction and Western blot analysis of RNA and protein extracts from different breast cancer cell lines confirmed the presence of endogenous coding SRA isoforms and their corresponding proteins. Our results demonstrate that full-length SRA-RNAs likely to encode stable proteins are widely expressed in breast cancer cell lines. PMID- 12565892 TI - Functional characterization of two variant human GSTO 1-1s (Ala140Asp and Thr217Asn). AB - Glutathione-S-transferase class Omega (GSTO 1-1) belongs to a new subfamily of GSTs, which is identical with human monomethylarsonic acid (MMA(V)) reductase, the rate limiting enzyme for biotransformation of inorganic arsenic, environmental carcinogen. Recombinant GSTO 1-1 variants (Ala140Asp and Thr217Asn) were functionally characterized using representative substrates. No significant difference was observed in GST activity towards 1-chloro-2,4-dinitrobenzene, whereas thioltransferase activity was decreased to 75% (Ala140Asp) and 40% (Thr217Asn) of the wild-type GSTO 1-1. For MMA(V) reductase activity, the Ala140Asp variant exhibited similar kinetics to wild type, while the Thr217Asn variant had lower V(max) (56%) and K(m) (64%) values than the wild-type enzyme. The different activities of the enzyme variants may influence both the intracellular thiol status and arsenic biotransformation. This can help explain the variation between individuals in their susceptibility to oxidative stress and inorganic arsenic. PMID- 12565893 TI - The brain-specific actin-related protein ArpN alpha interacts with the transcriptional co-repressor CtBP. AB - Actin-related protein (Arp) is found in many chromatin remodeling and histone acetyltransferase complexes. We previously identified ArpN alpha as an isoform of ArpN beta/BAF53, which is included in mammalian SWI/SNF chromatin remodeling complex, and showed that ArpN alpha is a potential component of the complex. Although it has a structure highly similar to ArpN beta/BAF53, ArpN alpha is expressed exclusively in brain and in neural differentiated embryonal carcinoma cells. Since ArpN alpha possesses a region that shows low similarity to ArpN beta/BAF53, we hypothesized that proteins interacting with this region contribute to the ArpN alpha-specific function in brain. Here we showed that ArpN alpha, but not ArpN beta/BAF53, interacts with the transcriptional co-repressor CtBP (C terminal binding protein). Transactivation by the SWI/SNF complex and glucocorticoid receptor was repressed by the CtBP in the presence of ArpN alpha. These findings suggest that SWI/SNF complex containing ArpN alpha might regulate certain genes involved in brain development and/or its function differently from SWI/SNF complex containing ArpN beta/BAF53. PMID- 12565894 TI - The signal peptide NPFSD fused to ricin A chain enhances cell uptake and cytotoxicity in Candida albicans. AB - Microorganisms possess stringent cell membranes which limit the cellular uptake of antimicrobials. One strategy to overcome these barriers is to attach drugs or research reagents to carrier peptides that enter cells by passive permeation or active uptake. Here the short endocytosis signal peptide NPFSD was found to efficiently deliver both FITC and GFP into Saccharomyces cerevisiae and Candida albicans with uptake into the majority of cells in a population. The NPFSD signal is itself non-toxic, but when fused to the ricin A chain toxin (RTA) the peptide enhanced both cell uptake and toxicity against C. albicans, which like other yeasts is resistant to naked RTA. Cell entry required at least 1 h incubation, temperatures above 4 degrees C, and an energy source, and uptake was out-competed with free peptide. Therefore, the NPFSD peptide can carry a range of compounds into yeasts and this delivery route holds promise to enhance the activity of antifungals. PMID- 12565895 TI - A novel single-stranded DNA enzyme expression system using HIV-1 reverse transcriptase. AB - In this study, we exploited a DNA enzyme expression system using the mechanism of HIV-1 reverse transcription in vitro. HIV-1 reverse transcription is initiated when its cognate primer tRNA (Lys-3) binds to the primer binding site (PBS) of the viral RNA template. Therefore, this RNA contains the HIV-1 PBS, the DNA enzyme, and a tRNA (Lys-3) at the 3(')-end of its RNA transcript, such that a single-stranded DNA (ssDNA) is synthesized by the HIV-1 reverse transcriptase. We constructed RNA expression vectors including the HIV-1 PBS, the DNA enzyme, and either a native tRNA (Lys-3) or one of two truncated tRNAs (Lys-3), Delta tRNA (Lys-3) and Delta Delta tRNA (Lys-3). The reactions of the pVAX1-Dz-tRNA (Lys-3), pVAX1-Dz-Delta tRNA (Lys-3), and pVAX1-Dz-Delta Delta tRNA (Lys-3) vectors with T7 RNA polymerase in vitro gave the corresponding RNAs. The liberated RNAs were treated with HIV-1 reverse transcriptase (HIV-1 RT) in vitro, which yielded the corresponding ssDNA. The cleavage assay results demonstrated that the expressed DNA enzyme has cleavage ability against the target sequence. Thus, we have found a new DNA enzyme oligonucleotide expression system using the HIV-1 reverse transcriptase in vitro. PMID- 12565896 TI - Homocysteine inhibits the proliferation and invasive potential of HT-1080 human fibrosarcoma cells. AB - The impairment of homocysteine metabolism has been related to several disorders and diseases. Recently, homocysteine has been shown to inhibit key steps of angiogenesis, including endothelial cell proliferation, invasion, and remodeling of the extracellular matrix. Since these are also key steps in tumor invasion and metastasis, it can be hypothesized that homocysteine can also interfere in these processes. Therefore, we studied the effects of homocysteine on tumor proliferation and invasion, as well as on urokinase, a key extracellular matrix degrading protease, using a model human tumor cell line. This study demonstrates that, in fact, homocysteine inhibits HT-1080 proliferation and invasion, and is a potent inhibitor of tumor cell urokinase expression. PMID- 12565897 TI - Purification of a trypsin-stable lectin with antiproliferative and HIV-1 reverse transcriptase inhibitory activity. AB - A lectin, with a molecular mass of approximately 60 kDa and two different subunits exhibiting an N-terminal sequence manifesting considerable homology to phytohemagglutinin from Phaseolus species, was isolated from the ground bean (Vigna sesquipedalis cv ground bean). The lectin was unique in hemagglutinating activity was inhibited by polygalacturonic acid and not by galacturonic acid and other simple monosaccharides. The lectin was isolated by affinity chromatography on Affi-gel blue gel, ion exchange chromatography by fast protein liquid chromatography (FPLC) on Mono S, and gel filtration by FPLC on Superdex 75. It was adsorbed on both Affi-gel blue gel and Mono S. Ground bean lectin exhibited mitogenic activity on murine splenocytes with the maximal response achieved at a concentration of 156 nM, as similar to the dose required for Con A. The viability of hepatoma (HepG2), leukemia (L1210), and leukemia (M1) cells was reduced in the presence of ground bean lectin, which also exerted an inhibitory activity toward HIV-1 reverse transcriptase IC(50) of 73 microM. The hemagglutinating activity of the lectin was unaffected by trypsinization and the presence of a number of divalent cations, but was augmented by 500 mM K(+) ions. The activity was unstable above 40 degrees C although some activity remained after heating and at 100 degrees C for 30s. PMID- 12565898 TI - Redox properties of the couples compound I/compound II and compound II/native enzyme of human myeloperoxidase. AB - Myeloperoxidase (MPO) is an important component of the neutrophil's antimicrobial armory and has been implicated in promoting tissue damage in numerous inflammatory diseases. For the first time the standard reduction potential of the redox couple compound II/native enzyme has been determined to be (0.97+/-0.01)V at pH 7.0 and 25 degrees C. This was achieved by rapid mixing of preformed compound II with either tyrosine or nitrite by using the sequential-mixing stopped-flow technique and measuring spectrophotometrically the concentrations of the reacting species and products at equilibrium. Using the recently determined standard reduction potential for the couple compound I/native enzyme (1.16 V), the reduction potential of the couple compound I/compound II was calculated to be 1.35 V at pH 7 and 25 degrees C. These data reveal substantial differences between the two known heme peroxidase superfamilies and reflect the dramatic differences observed in the oxidisability of substrates by the MPO redox intermediates compound I and compound II. PMID- 12565899 TI - Conservation and cloning of CYP51: a sterol 14 alpha-demethylase from Mycobacterium smegmatis. AB - The genetic locus encoding cytochrome P450 51 (CYP51; P450(14DM)) in Mycobacterium smegmatis is described here together with confirmation of activity in lanosterol 14 alpha-demethylation. The protein bound azole antifungals with high affinity and the rank order based on affinity matched the ranked order for microbiological sensitivity of the organism, thus supporting a possible role for CYP51 as a target in the antimycobacterial activity of these compounds. Non saponifiable lipids were extracted from the bacteria grown on minimal medium. Unlike a previous report using growth on complex medium, no cholesterol was detected in two strains of M. smegmatis, but a novel lipid was detected. The genetic locus of CYP51 is discussed in relation to function; it is conserved as part of a putative operon in M. smegmatis, Mycobacterium tuberculosis, Mycobacterium avium, and Mycobacterium bovis and consists of six open-reading frames including two CYPs and a ferredoxin under a putative Tet-R regulated promoter. PMID- 12565900 TI - Insulin or bFGF and C2 ceramide increase newborn rat retinal ganglion cell survival rate. AB - Treatment of RGCs with insulin or C2 ceramide alone increased survival rate by 30%. Adding both insulin and C2 ceramide increased survival rate by 80%. Protein phosphatase 2A (PP2A) inhibitor okadaic acid (OA) eliminated the effect of C2 ceramide, but not that of insulin. Protein kinase inhibitor K252a decreased the effect of C2 ceramide in a dose-dependent manner, but the effect of insulin was not changed. Treatment of RGCs with bFGF increased survival rate by 36%. Adding both bFGF and C2 ceramide increased survival rate by 102%. OA did not alter the effect of bFGF, whereas K252a increased survival rate in a dose-dependent manner. Inhibition of C2 ceramide by OA suggests that PP2A activation is involved in its pathway, whereas PP2A is not involved in the insulin- and bFGF-activated pathway. Elimination of the effect of C2 ceramide by K252a suggests that sphingomyelin cycle activation is mediated by a protein kinase not important in the insulin activated pathway. Moreover, the increased effect of bFGF and dose-dependently decreased effect of C2 ceramide by K252a suggest that different protein kinases are important in bFGF- and ceramide-mediated enhancement of RGC survival rate. PMID- 12565901 TI - The effect of oxidative stress on histone acetylation and IL-8 release. AB - Acetylation of histone residues regulates the expression of inflammatory genes and is controlled by the activities of histone acetyltransferases (HAT) and histone deacetylases (HDAC). Analysis of histone acetylation in human cells is limited by the large numbers needed to perform activity assays or Western blotting. We have used flow cytometry to investigate changes in HAT and HDAC activities at the single cell level and to investigate the effect of hydrogen peroxide (H(2)O(2)) on histone H4 acetylation and cell-cycle progression. Using an anti-acetylated histone H4 antibody we show that H(2)O(2) induced a time dependent increase in histone acetylation that was maintained for 12h. This was associated with increased IL-8 production. H(2)O(2) also affected cell-cycle progression. HAT activity was found to be highest in G2/M and equivalent in G0/G1 and S phases of the cell cycle. These data show that detection of acetylated histone residues at the single cell level using FACs may be a powerful new tool for the analysis of modulation of cell proliferation and gene transcription. PMID- 12565902 TI - Reduced expression of PGC-1 and insulin-signaling molecules in adipose tissue is associated with insulin resistance. AB - Peroxisome proliferator-activated receptor gamma (PPAR gamma) co-activator 1 (PGC 1) regulates glucose metabolism and energy expenditure and, thus, potentially insulin sensitivity. We examined the expression of PGC-1, PPAR gamma, insulin receptor substrate-1 (IRS-1), glucose transporter isoform-4 (GLUT-4), and mitochondrial uncoupling protein-1 (UCP-1) in adipose tissue and skeletal muscle from non-obese, non-diabetic insulin-resistant, and insulin-sensitive individuals. PGC-1, both mRNA and protein, was expressed in human adipose tissue and the expression was significantly reduced in insulin-resistant subjects. The expression of PGC-1 correlated with the mRNA levels of IRS-1, GLUT-4, and UCP-1 in adipose tissue. Furthermore, the adipose tissue expression of PGC-1 and IRS-1 correlated with insulin action in vivo. In contrast, no differential expression of PGC-1, GLUT-4, or IRS-1 was found in the skeletal muscle of insulin-resistant vs insulin-sensitive subjects. The findings suggest that PGC-1 may be involved in the differential gene expression and regulation between adipose tissue and skeletal muscle. The combined reduction of PGC-1 and insulin signaling molecules in adipose tissue implicates adipose tissue dysfunction which, in turn, can impair the systemic insulin response in the insulin-resistant subjects. PMID- 12565903 TI - NF-kappa B p65 transactivation domain is involved in the NF-kappa B-inducing kinase pathway. AB - NF-kappa B-inducing kinase (NIK) is involved in the signal transduction pathway leading to the NF-kappa B activation. In this report, we demonstrate that the NIK mediated NF-kappa B activation involves the transactivation (TA) domain of p65 subunit of NF-kappa B and the nuclear translocation of IKK alpha. By using luciferase assay, we found that both IKK alpha and IKK beta could activate NF kappa B in synergy with NIK. Interestingly, although IKK beta stimulated the NIK mediated I kappa B degradation, IKK alpha stimulated the action of NF-kappa B without enhancing the I kappa B degradation. By using heterologous transactivation system with Gal4 DNA-binding domain in fusion with various portions of p65 TA domain, we found that the transactivation domain 1 (TA1) of p65 serves as the direct target for the NIK-IKK alpha cascade and that the serine residue at 536 within p65 TA1 is indispensable for this action. Furthermore, we found that this action of NIK depends on the energy-dependent action of Ras related protein (Ran) since the dominant negative mutant of Ran (RanQ69L) inhibited the transcriptional activity of p65 by preventing the nuclear import of IKK alpha. PMID- 12565904 TI - Chemical modification of arginine alleviates the decline in activity during catalysis of spinach Rubisco. AB - Arginine residues of spinach ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) were chemically modified with phenylglyoxal (PhG). PhG inactivated Rubisco with a half-time of 20-25 min. An inclusion of a catalytic product, 3 phosphoglycerate (PGA), protected Rubisco from inactivation and delayed the half time to 60-90 min. Peptide mapping and sequencing of Rubisco modified for 60 min with radiolabeled PhG in the presence of 10mM PGA revealed that Arg187, Arg258, and Arg431 of the large subunit were modified. The extent and rate of the decline in activity during catalysis (fallover phenomenon) were reduced by the modification. This is the first report identifying PhG-modified arginine residues and to demonstrate the effect of the modification of arginine residues on the kinetics of fallover. PMID- 12565905 TI - Homozygous deletions and point mutations of the Rit1/Bcl11b gene in gamma-ray induced mouse thymic lymphomas. AB - Allelic loss (LOH) mapping and sequence analysis were conducted for gamma-ray induced mouse thymic lymphomas and a novel tumor suppressor gene, Rit1/Bcl11b, on chromosome 12 was isolated. Bi-allelic changes were found in 17 of the 66 p53 proficient lymphomas with Rit1 LOH but in only 2 of the 54 p53-deficient lymphomas. This suggests an association between the presence of functional p53 and inactivation of the Rit1 gene in the lymphoma development. Introduction of Rit1 into HeLa cells lacking Rit1 expression suppressed cell growth. These results indicate that loss-of-function mutations of Rit1 contribute to mouse lymphomagenesis and possibly to human cancer development. PMID- 12565906 TI - Angiopoietin-like protein 3, a hepatic secretory factor, activates lipolysis in adipocytes. AB - Our previous work identified a genetic mutation in the gene encoding angiopoietin like protein 3 (Angptl3) in KK/Snk mice (previously KK/San), a mutant strain of KK obese mice. KK/Snk had significantly lower plasma triglyceride and free fatty acid (FFA) than KK mice. Human ANGPTL3 treatment increased both plasma triglyceride and FFA. ANGPTL3 inhibited the activity of lipoprotein lipase, which accounted for the increase of plasma triglyceride. The mechanism how ANGPTL3 affects plasma FFA has not been known. The current study reveals that ANGPTL3 targets on adipose cells and induces lipolysis. Both plasma FFA and glycerol decreased in KK/Snk and increased by the treatment of human ANGPTL3. Specific bindings of ANGPTL3 to adipose cells were shown using fluorescence-labeled protein visually and 125I-labeled protein by the binding analysis. Furthermore, ANGPTL3 activated the lipolysis to stimulate the release of FFA and glycerol from adipocytes. We conclude that ANGPTL3 is a liver-derived lipolytic factor targeting on adipocyte. PMID- 12565907 TI - In situ and in vitro evidence for DCoH/HNF-1 alpha transcription of tyrosinase in human skin melanocytes. AB - Human epidermal melanocytes hold the full capacity for autocrine de novo synthesis/regulation/recycling of the essential cofactor 6-tetrahydrobiopterin (6BH(4)) for conversion of L-phenylalanine via phenylalanine hydroxylase to L tyrosine and for production of L-Dopa via tyrosine hydroxylase to initiate both pigmentation and catecholamine synthesis in these neural crest-derived cells. Earlier we have demonstrated pterin-4a-carbinolamine dehydratase (PCD) mRNA and enzyme activities in epidermal melanocytes and keratinocytes. This protein dimerises also the transcription factor hepatocyte nuclear factor 1 (HNF-1), leading to activation of multiple genes. This study demonstrates for the first time DCoH/HNF-1 alpha expression and transcriptional activity in human epidermal melanocytes in vitro and in situ and identified tyrosinase, the key enzyme for pigmentation, as a new transcriptional target. Specific binding of DCoH/HNF-1 complex to the human tyrosinase promoter was confirmed by gel shift analysis. These results provide a novel mechanism in the regulation of skin pigmentation. PMID- 12565909 TI - Daytime predictors of sleep disordered breathing in children and adolescents with neuromuscular disorders. AB - Sleep disordered breathing with or without nocturnal hypercapnic hypoventilation is a common complication of respiratory muscle weakness in childhood neuromuscular disorders. Nocturnal hypercapnic hypoventilation as a sign of respiratory muscle fatigue, portends a particularly poor prognosis. We aimed at identifying daytime predictors of sleep disordered breathing at its onset and sleep disordered breathing with nocturnal hypercapnic hypoventilation. Forty-nine children and adolescents (11.3+/-4.4 years) with progressive neuromuscular disorders were studied with inspiratory vital capacity, peak inspiratory pressure, arterial blood gases, polysomnography, and a ten-item symptoms questionnaire. Daytime respiratory function was prospectively compared with polysomnographic variables. Sleep disordered breathing was found in 35/49 patients (71%). Twenty-four (49%) had sleep disordered breathing with nocturnal hypercapnic hypoventilation. Inspiratory vital capacity and peak inspiratory pressure, but not symptom score, correlated with sleep disordered breathing and severity of nocturnal hypercapnic hypoventilation. Sleep disordered breathing onset was predicted by inspiratory vital capacity<60% (sens. 97%, spec. 87%). Sleep disordered breathing with nocturnal hypercapnic hypoventilation was predicted by inspiratory vital capacity<40% (sens. 96%, spec. 88%) and PaCO(2)>40 mmHg (sens. 92%, spec. 72%,). Sleep disordered breathing can reliably be predicted from simple daytime respiratory function tests, which, if applied systematically, will improve recognition of nocturnal respiratory failure. PMID- 12565908 TI - Bone morphogenetic protein signaling in articular chondrocyte differentiation. AB - Articular chondrocytes progressively undergo dedifferentiation into a spindle shaped mesenchymal cellular phenotype in monolayers. Chondrocyte dedifferentiation is stimulated by retinoic acid. On the other hand, bone morphogenic proteins (BMPs) stimulate differentiation of chondrocytes. We examined the mechanism of effects of BMP in chondrocyte differentiation with use of a recombinant adenovirus vector system. Constitutively active forms of BMP type I receptors (BMPR-IA and BMPR-IB) and those of activin receptor-like kinase (ALK)-1 and ALK-2 maintained differentiation of chondrocytes in the presence of retinoic acid. The BMP receptor-regulated signaling substrates, Smad1/5, weakly induced chondrocyte differentiation; the effects of Smad1/5 were enhanced by BMP 7 treatment. Inhibitory Smad, Smad6, blocked increase of expression of chondrocyte markers by BMP-7 in a dose-dependent manner. SB202190, a p38 mitogen activated protein kinase inhibitor, inhibited this effect of BMP-7; however, since SB202190 suppressed phosphorylation of Smad1/5, this may be due to blockade of BMP receptor activation. These results together strongly suggest that induction of chondrocyte differentiation by BMP-7 is regulated by Smad pathways. PMID- 12565910 TI - Cardiac assessment in childhood carriers of Duchenne and Becker muscular dystrophies. AB - Cardiac disease in adult female carriers of the X-linked dystrophinopathies, Duchenne and Becker muscular dystrophies, is a well-recognised entity. A single study has reported a 15% incidence of cardiac abnormalities in female carriers under 16 years. Our study aims, clinically and with electrocardiograph and echocardiograph, to determine the incidence of cardiac abnormality in young girls who are proven carriers of X-linked dystrophinopathies. Twenty-three girls aged 6.2-15.9 years were assessed. All had normal cardiac examination. None had electrocardiograph abnormalities consistent with dystrophic cardiomyopathy. Left ventricular fractional shortening ranged from 33 to 55% (normal>28%). Septal thickness, posterior wall thickness and wall thickness ratio were within normal limits. No cardiac abnormalities have been demonstrated in young girls who are proven carriers of X-linked dystrophinopathies in our study. This has important implications for planning timing of carrier determination and cardiac assessment. PMID- 12565911 TI - Recessive POLG mutations presenting with sensory and ataxic neuropathy in compound heterozygote patients with progressive external ophthalmoplegia. AB - Autosomal recessive progressive external ophthalmoplegia is a mitochondrial disease characterized by accumulation of multiple large-scale deletions of mitochondrial DNA. We previously reported missense mutations in POLG, the gene encoding the mitochondrial DNA polymerase gamma in two nuclear families compatible with autosomal recessive progressive external ophthalmoplegia. Here, we report a novel POLG missense mutation (R627W) in a sporadic patient and we provide genetic support that all these POLG mutations are actually causal and recessive. The novel patient presented with sensory ataxic neuropathy and has the clinical triad of sensory ataxic neuropathy, dysarthria and ophthalmoparesis (SANDO). This is the first finding of a genetic cause of Sensory Ataxic Neuropathy, Dysarthria and Ophthalmoparesis and it implies that this disorder may actually be a variant of autosomal recessive progressive external ophthalmoplegia. Sensory neuropathy is the initial feature in Belgian compound heterozygote autosomal recessive progressive external ophthalmoplegia patients, all carrying the POLG A467T mutation, which occurs at a frequency of 0.6% in the Belgian population. PMID- 12565912 TI - Transcriptional activation of the utrophin promoter B by a constitutively active Ets-transcription factor. AB - Duchenne muscular dystrophy is an X-linked genetic disease caused by the absence of functional dystrophin. Pharmacological upregulation of utrophin, the autosomal homologue of dystrophin, offers a potential therapeutic approach to treat Duchenne patients. Full-length utrophin mRNA is transcribed from two alternative promoters, called A and B. In contrast to the utrophin promoter A, little is known about the factors regulating the activity of the utrophin promoter B. Computer analysis of this second promoter revealed the presence of several conserved binding motives for Ets-transcription factors. Using electrotransfer of cDNA into mouse muscles, we demonstrate that a genetically modified beta-subunit of the Ets-transcription factor GA-binding protein potently activates a utrophin promoter B reporter construct in innervated muscle fibers in vivo. These results make the GA-binding protein and the signaling cascade regulating its activity in muscle cells, potential targets for the pharmacological modulation of utrophin expression in Duchenne patients. PMID- 12565914 TI - Childhood chronic inflammatory demyelinating polyneuropathy with central nervous system demyelination resembling multiple sclerosis. AB - Central nervous system demyelination has been described in adults but not in children with chronic inflammatory demyelinating polyneuropathy. We describe a patient with clinical and electrophysiological features consistent with chronic inflammatory demyelinating polyneuropathy who presented at age 5 with an intramedullary spinal cord tumor-like lesion and at age 8, represented with cerebral and spinal demyelinating lesions. Her clinical course and magnetic resonance imaging features were atypical for multiphasic disseminated encephalomyelitis and indistinguishable from multiple sclerosis. To our knowledge, this association has not been previously described in the English literature in childhood. PMID- 12565913 TI - Principal mutation hotspot for central core disease and related myopathies in the C-terminal transmembrane region of the RYR1 gene. AB - The congenital myopathies are a group of disorders characterised by the predominance of specific histological features observed in biopsied muscle. Central core disease and nemaline myopathy are examples of congenital myopathies that have specific histological characteristics but significantly overlapping clinical pictures. Central core disease is an autosomal dominant disorder with variable penetrance which has been linked principally to the gene for the skeletal muscle calcium release channel (RYR1). Two recent reports have identified the 3' transmembrane domain of this gene as a common site for mutations. Two other studies have reported single families that have features of both central core disease and nemaline myopathy (core/rod disease) caused by mutations in RYR1. Screening of the 3' region (exons 93-105) of the RYR1 gene for mutations in 27 apparently unrelated patients with either central core disease or core/rod disease by single strand conformation polymorphism analysis and DNA sequencing identified three described and nine novel mutations in 15 patients. PMID- 12565916 TI - 107th ENMC international workshop: the management of cardiac involvement in muscular dystrophy and myotonic dystrophy. 7th-9th June 2002, Naarden, the Netherlands. PMID- 12565915 TI - Autosomal dominant external ophthalmoplegia and bipolar affective disorder associated with a mutation in the ANT1 gene. AB - The authors report on a family with dominantly inherited progressive external ophthalmoplegia and a diagnostic and statistical manual (fourth revised edition) diagnosis of bipolar psychiatric disorder in several members. Skeletal muscle biopsy from the proposita showed decreased cytochrome c oxidase staining, several ragged-red fibers, and multiple mtDNA deletions. The authors identified a missense mutation (leucine 98-->proline) in the adenine nucleotide translocator 1 gene. The presence of bipolar affective disorder expands the phenotype of adenine nucleotide translocator 1 allelic variants. PMID- 12565917 TI - 103rd ENMC international workshop: designing rational therapy of SMA based on the understanding of its pathophysiology, 18-20 January 2002, Naarden, The Netherlands. PMID- 12565918 TI - 93rd ENMC international workshop: non-5q-spinal muscular atrophies (SMA) - clinical picture (6-8 April 2001, Naarden, The Netherlands). PMID- 12565919 TI - Muscular dystrophy campaign sponsored workshop: recommendation for respiratory care of children with spinal muscular atrophy type II and III. 13th February 2002, London, UK. PMID- 12565921 TI - Myxovirescin analogues via macrocyclic ring-closing metathesis. AB - A short, efficient route has been developed to analogues of myxovirescin using ring-closing metathesis whereby the antibacterial activity has been retained. PMID- 12565922 TI - Chemoenzymatic synthesis of high-mannose type HIV-1 gp120 glycopeptides. AB - A chemoenzymatic approach to the synthesis of glycoforms of HIV-1 gp120 glycopeptides is described. Thus, the high-mannose type glycopeptides [gp120 (336 342)] containing Man(9), Man(6) and Man(5) moieties, respectively, were synthesized in satisfactory yields via transglycosylation to the acetylglucosaminyl peptide, using the recombinant Arthrobacter Endo-beta-N-acetyl glucosaminidase (Endo-A) as the key enzyme. PMID- 12565923 TI - Solid-phase synthesis of cyclic RGD-furanoid sugar amino acid peptides as integrin inhibitors. AB - The solid-phase synthesis of cyclic RGD peptides containing either one or two furanoid sugar amino acids (SAAs) is reported. Using a cyclization-cleavage approach five peptides were successfully assembled and consecutively tested on their ability to bind to the integrin receptors alpha(v)beta(3) and alpha(IIb)beta(3). The cyclic tetrapeptide c[RGD-SAA] (1) showed the most promising activity in an inhibition assay with an IC(50) of 1.49 microM for the alpha(v)beta(3) receptor and 384 nM for the alpha(IIb)beta(3) receptor. PMID- 12565924 TI - 2,5-Bis-(2-hydroxybenzoylamino)pentanoic acid, a salicylic acid-metabolite isolated from chicken: characterization and independent synthesis. AB - From excreta of chickens that had been treated with sodium salicylate, a new compound was detected and identified as a double conjugated ornithine metabolite. The structural assignment of this metabolite was further confirmed by an independent efficient 3-step synthesis from ornithine. PMID- 12565925 TI - Stereochemical studies on phosphopantothenoylcysteine decarboxylase from Escherichia coli. AB - Phosphopantothenoylcysteine decarboxylase catalyzes the decarboxylation of 4' phosphopantothenoylcysteine (2) to form 4'-phosphopanthetheine (3), an intermediate in the biosynthesis of Coenzyme A. In this study we investigated the stereochemistry of this reaction. Our results show that the decarboxylation proceeds with retention of stereochemistry, and that the pro-R proton at C(beta) of the cysteine moiety of 2 is removed during a reversible oxidation of the thiol to a thioaldehyde intermediate. PMID- 12565926 TI - Synthesis and GABA(A) receptor activity of a 6,19-oxido analogue of pregnanolone. AB - 3 alpha-Hydroxy-6,19-oxidopregn-4-ene-20-one (4) was prepared in seven steps from pregnanolone acetate. At 0.1 microM concentration 4 significantly increased GABA induced (36)Cl(-) influx in hamster cerebral cortex synaptoneurosomes while at 20 mg/kg it decreased the percentage of hamsters showing seizures induced by 3 mercaptopropionic acid. PMID- 12565927 TI - Imidazopyrimidines, potent inhibitors of p38 MAP kinase. AB - The MAP kinase p38 is implicated in the release of the pro-inflammatory cytokines TNF-alpha and IL-1 beta. Inhibition of cytokine release may be a useful treatment for inflammatory conditions such as rheumatoid arthritis and Crohn's disease. A novel series of imidazopyrimidines have been discovered that potently inhibit p38 and suppress the production of TNF-alpha in vivo. PMID- 12565928 TI - [3H]-methoxymethyl-MTEP and [3H]-methoxy-PEPy: potent and selective radioligands for the metabotropic glutamate subtype 5 (mGlu5) receptor. AB - The design, synthesis, and characterization of two potent, non-competitive radioligands, [3H]-methoxymethyl-MTEP and [3H]-methoxy-PEPy, that are selective for the mGlu5 receptor are described. PMID- 12565929 TI - Structure-activity relationships of novel anti-malarial agents: part 5. N-(4 acylamino-3-benzoylphenyl)-[5-(4-nitrophenyl)-2-furyl]acrylic acid amides. AB - We have developed the [5-(4-nitrophenyl)-2-furyl]acrylic acid substituted benzophenone 4g as a novel lead for anti-malarial agents. Here, we demonstrated that the acyl residue at the 2-amino group of the benzophenone core structure has to be a phenylacetic acid substructure substituted in its para-position with methyl or other substituents of similar size. The trifluoromethyl substituted derivative displayed an IC(50) of 47 nM against the multi-drug resistant Plasmodium falciparum strain Dd2. PMID- 12565930 TI - Nociceptin/Orphanin FQ(1-13)NH2 analogues modified in the Phe1-Gly2 peptide bond. AB - The synthesis and pharmacological activity of novel nociceptin/orphanin FQ (N/OFQ) analogues modified in the Phe(1)-Gly(2) peptide bond are reported. The aim of the present work was to elucidate the importance of this peptide bond for the N/OFQ receptor (NOP) interaction. Our study indicates that the first peptide bond in N/OFQ is important but not crucial for interaction with the N/OFQ receptor; for instance, substitution with a methyleneoxy bond generates an agonist derivative just 3-fold less potent than the reference compound. PMID- 12565931 TI - Nonbenzamidine tetrazole derivatives as factor Xa inhibitors. AB - Factor Xa (fXa) is an important serine protease that holds the central position linking the intrinsic and extrinsic activation mechanisms in the blood coagulation cascade. Therefore, inhibition of fXa has potential therapeutic applications in the treatments of both arterial and venous thrombosis. Herein we describe a series of tetrazole fXa inhibitors containing benzamidine mimics as the P(1) substrate, of which the aminobenzisoxazole moiety was found to be the most potent benzamidine mimic. SR374 (12) inhibits fXa with a K(i) value of 0.35 nM and is very selective for fXa over thrombin and trypsin. PMID- 12565932 TI - Synthesis and In vivo anti-inflammatory activity of long-chain 2-amino-alcohols. AB - The synthesis of optically pure long-chain 2-amino-alcohols and 1-O-dodecyl-2 deoxy-2-amino-sn-glycerol was carried out starting from L- or D-Boc-Ser(OBn)-ol by oxidation and consecutive Wittig reaction or etherification reaction. 2-Amino oleyl alcohol was synthesized by reduction of the corresponding 2-amino-oleic acid. All the long chain amino-alcohols presented interesting inhibition of carrageenin-induced paw edema in rats (ED(50) from 0.017 to 0.010 mmol/kg). PMID- 12565933 TI - Discovery of a novel series of 6-azauracil-based thyroid hormone receptor ligands: potent, TR beta subtype-selective thyromimetics. AB - In this communication, we wish to describe the discovery of a novel series of 6 azauracil-based thyromimetics that possess up to 100-fold selectivities for binding and functional activation of the beta(1)-isoform of the thyroid receptor family. Structure-activity relationship studies on the 3,5- and 3'-positions provided compounds with enhanced TR beta affinity and selectivity. Key binding interactions between the 6-azauracil moiety and the receptor have been determined through of X-ray crystallographic analysis. PMID- 12565934 TI - Evaluation of vinylsulfamides as sulfhydryl selective alkylation reagents in protein modification. AB - The preparation of several model vinylsulfamides is described. Their excellent selective reactivity towards sulfhydryl group with regards to amino group has been demonstrated by the kinetics study between a model vinylsulfamide and cysteine or lysine at different pHs. PMID- 12565935 TI - Synthesis and evaluation of delta-lactams (piperazones) as elastase inhibitors. AB - A series of monocyclic delta-lactams (piperazones) was prepared and analysed as inhibitors of porcine pancreatic elastase and human neutrophil elastase. PMID- 12565936 TI - Synthesis of labelled PNA oligomers by a post-synthetic modification approach. AB - The preparation of t-butoxycarbonyl (Boc)-protected O(4)-(o-nitrophenyl) thymine peptide nucleic acid (PNA) monomer is described. This PNA monomer was incorporated into PNA oligomer sequences. The post-synthetic modification of the oligomers to yield fluorescently-labelled PNA oligomers was studied before and after the removal of the protecting groups. In both cases, the desired fluorescently-labelled PNA oligomer was obtained in good yields. PMID- 12565937 TI - Synthesis of antioxidant propyl gallate using tannase from Aspergillus niger van Teighem in nonaqueous media. AB - Tannase from Aspergillus niger van Teighem has been used for synthesis of food additive antioxidant propyl gallate by direct transesterification of tannic acid. The optimized yield of 86% was obtained by using simultaneously pH tuned enzyme, immobilized on Celite and using the right amount of water in the non aqueous media. PMID- 12565938 TI - Novel thieno oxazine analogues as antihyperglycemic and lipid modulating agents. AB - A series of phenyl acetic acid and alpha-hydroxy propionic acid derivatives were synthesized. In vivo studies of the compounds indicated compound 2c as the most potent in one of the series, which has both glucose and lipid lowering properties. The syntheses and biological studies have been discussed. PMID- 12565939 TI - SLV310, a novel, potential antipsychotic, combining potent dopamine D2 receptor antagonism with serotonin reuptake inhibition. AB - In this paper, SLV310 is presented as a novel, potential antipsychotic displaying the interesting combination of potent dopamine D(2) receptor antagonism and serotonin reuptake receptor inhibition in one molecule. As such, SLV310 could be useful in treating a broad range of symptoms in schizophrenia. This paper describes the structure-activity relationship in a series of compounds leading to SLV310 (6b, 2-[4-[4-(5-fluoro-1H-indol-3-yl)-3,6-dihydro-2H-pyridin-1-yl]-butyl] phthalimide) together with pharmacological data showing the unique profile of this compound. PMID- 12565940 TI - Zinc(II)-mediated inhibition of a ribonuclease by an N-hydroxyurea nucleotide. AB - The inhibition of ribonuclease Bi by 3'-N-hydroxyurea-3'-deoxythymidine 5' phosphate is enhanced by 30-fold in the presence of Zn(2+). Thus, an N hydroxyurea nucleotide can recruit Zn(2+) to inhibit the enzymatic activity of a ribonuclease. This result engenders a general strategy for the inhibition of non metalloenzymes by metal complexes. PMID- 12565941 TI - 2,3-Diarylbenzopyran derivatives as a novel class of selective cyclooxygenase-2 inhibitors. AB - A new series of cyclooxygenase-2(COX-2) inhibitors with naturally occurring flavone as the main skeleton has been synthesized and their biological activities were evaluated for cyclooxygenase inhibitory activity. Rational structural modifications were applied to potent COX-2 inhibitors to obtain the desired pharmacokinetic profiles for improved oral anti-inflammatory activity. PMID- 12565942 TI - QSAR study on solubility of alkanes in water and their partition coefficients in different solvent systems using PI index. AB - The aqueous solubility (S(w)) of liquids and solids, expressed as log S(w) as well as their partition coefficients in different solvent systems viz. P(oct) (partition coefficient in octanol-water), P(16) (partition coefficient in water hexadecane), P(alk) (partition coefficient in water-alkane), and P(cyc) (partition coefficient in water-cyclohexane), and aqueous solubility (S(w)) have been estimated using the PI (Padmakar-Ivan) index and the results compared with those obtained using the widely used Wiener index (W). Regression analysis of the data using n-alkanes show that the PI index gives better results than the W index. PMID- 12565943 TI - A series of 2(Z)-2-benzylidene-6,7-dihydroxybenzofuran-3[2H]-ones as inhibitors of chorismate synthase. AB - A series of 2(Z)-2-benzylidene-6,7-dihydroxybenzofuran-3[2H]-ones was identified as potent inhibitors of bacterial chorismate synthase. The 2'-hydroxy-4'-pentoxy analogue 33 is a potent inhibitor of Streptococcus pneumoniae chorismate synthase. PMID- 12565944 TI - 1,3,4 Trisubstituted pyrrolidine CCR5 receptor antagonists bearing 4 aminoheterocycle substituted piperidine side chains. AB - A new class of 4-(aminoheterocycle)piperidine derived 1,3,4 trisubstituted pyrrolidine CCR5 antagonists is reported. Compound 4a is shown to have good binding affinity (1.8 nM) and antiviral activity in PBMC's (IC(95)=50 nM). Compound 4a also has improved PK properties relative to 1. PMID- 12565945 TI - Novel antifungal beta-amino acids: synthesis and activity against Candida albicans. AB - A series of novel beta-amino acids has been synthesized and tested for their in vitro antifungal activity against Candida albicans. A steep SAR was observed. beta-Amino acid 21 (BAY 10-8888/PLD-118) revealed the most favourable activity tolerability profile and was selected for clinical studies as a novel antifungal for the oral treatment of yeast infections. PMID- 12565946 TI - Discovery of orally bioavailable NK1 receptor antagonists. AB - Benzyloxyphenethylpiperazines are a new class of high affinity NK1 receptor antagonists. Oral bioavailability and selectivity can be fine tuned by the nature of the substituents on the basic nitrogen atom. Addition of substituents with a carboxylic acid group led to very selective and orally active NK1 antagonists free of interaction with L-type calcium channels. PMID- 12565947 TI - Characterization of the mechanism of anticonvulsant activity for a selected set of putative AMPA receptor antagonists. AB - A selected set of 1-aryl-7,8-methylenedioxy-2,3-benzodiazepin-4-ones and their analogues were evaluated for their ability to bind the competitive and noncompetitive sites of the AMPA receptors complex as well as to the glycine site of the NMDA receptors. The results put in evidence that most of the test compounds, despite a close structural similarity with GYKI 52466, possess a significantly different pharmacological profile. PMID- 12565948 TI - Modelling of carbonic anhydrase inhibitory activity of sulfonamides using molecular negentropy. AB - The present paper deals with the modelling of carbonic anhydrase inhibitory activity of sulfonamides using molecular negentropy (N). Excellent results are obtained in multiple regression analysis upon introduction of indicator parameters. The results are critically discussed on the basis of statistical data obtained from regression analysis. PMID- 12565949 TI - Synthesis and antitumor activity of novel C-8 ester derivatives of leinamycin. AB - A novel series of C-8 ester derivatives of leinamycin are described. Condensation of N-substituted amino acids or carboxylic acids containing polyether moiety with leinamycin resulted in the C-8 ester derivatives with good antitumor activity in several experimental models. Among these derivatives, compound 4e, which has five ethylene glycol ether units in the C-8 acyl group, showed potent antitumor activity against human tumor xenograft. Combination with the modification of the dithiolanone moiety was applied to these C-8 ester derivatives and some of them also showed good antitumor activity. PMID- 12565950 TI - Synthesis and preliminary cytotoxicity of nitrogen mustard derivatives of distamycin A. AB - Distamycin and nitrogen mustard conjugates, in which the nitrogen mustard unit was coupled to the C-terminus of the pyrrole, were synthesized. The switching of the nitrogen mustard unit from the N-terminus to the C-terminus did not compromise the compound's cytotoxicity. Compound 3, bearing three pyrrole units, was highly toxic to human K562 leukemia cells in vitro with an IC(50) value of 0.03 microM. Addition of a trans double bond to the molecule had little effects on cytotoxicity. PMID- 12565951 TI - Synthesis and biological evaluation of novel 1 beta-methylcarbapenems with isothiazoloethenyl side chains. AB - The synthesis of novel 1 beta-methylcarbapenems 1a,b bearing isothiazoloethenyl moieties at C-5 position of pyrrolidine ring and their biological evaluation are described. Both compounds showed potent and well-balanced antibacterial activity as well as high stability to DHP-I. Especially, 5-isothiazole derivative 1a exhibited excellent DHP-I stability and advanced pharmacokinetics profiles, compared to 5-isoxazole derivative 2, imipenem, and meropenem. PMID- 12565952 TI - p38 Inhibitors: piperidine- and 4-aminopiperidine-substituted naphthyridinones, quinolinones, and dihydroquinazolinones. AB - We have synthesized a series of C7-piperidine- and 4-aminopiperidine-substituted naphthyridinones, quinolinones, and dihydroquinazolinones that are highly potent inhibitors of both p38MAP kinase activity and TNF-alpha release. The 4 aminopentamethylpiperidine naphthyridinone 5, which was designed to block metabolism at major 'hot spots', combined excellent inhibitory potency with good oral bioavailability in the rat. PMID- 12565953 TI - Antitumour benzothiazoles. Part 20: 3'-cyano and 3'-alkynyl-substituted 2-(4' aminophenyl)benzothiazoles as new potent and selective analogues. AB - The synthesis of a new series of antitumour 2-(4-aminophenyl)benzothiazole analogues, substituted in the 3'-position by cyano or alkynyl groups, is described. Several of the analogues, notably the 5-fluorinated compounds 7c and 9c, were found to possess potent in vitro activity against MCF-7 and MDA 468 human cancer cell lines. More comprehensive in vitro analysis (NCI 60-cell line) established compound 7c as a particularly potent and selective 2-(4 aminophenyl)benzothiazole analogue. PMID- 12565954 TI - Synthesis and SAR of alpha-acylaminoketone ligands for control of gene expression. AB - A lead discovery library and a follow-up focused library of alpha acylaminoketones were designed based on known dibenzoylhydrazine ecdysone agonists, including GS(TM)-E. The compounds were assayed in mammalian cells expressing the ecdysone receptor from Bombyx mori for their ability to cause expression of a reporter gene downstream of an ecdysone response element. The most potent alpha-acylaminoketones were comparable to GS(TM)-E in this assay. PMID- 12565955 TI - 2-Phenylspiroindenes: a novel class of selective estrogen receptor modulators (SERMs). AB - A series of 2-phenylspiroindenes was prepared. The most active analogue (2) was found to be comparable in potency to raloxifene (1) as an estrogen receptor ligand. PMID- 12565956 TI - Inhibition of mast cell leukotriene release by thiourea derivatives. AB - Mast cell derived leukotrienes (LT's) play a vital role in pathophysiology of allergy and asthma. We synthesized various analogues of indolyl, naphthyl and phenylethyl substituted halopyridyl, thiazolyl and benzothiazolyl thioureas and examined their in vitro effects on the high affinity IgE receptor/FcERI-mediated mast cell leukotriene release. Of the 22 naphthylethyl thiourea compounds tested, there were seven active compounds and N-[1-(1-naphthyl)ethyl]-N'-[2-(ethyl-4 acetylthiazolyl)]thiourea (17 and 16) (IC(50)=0.002 microM) and N-[1-(1R) naphthylethyl]-N'-[2-(5-methylpyridyl)]thiourea (5) (IC(50)=0.005 microM) were identified as the lead compounds. Among the 11 indolylethyl thiourea compounds tested, there were seven active compounds and the halopyridyl compounds N-[2-(3 indolylethyl)]-N'-[2-(5-chloropyridyl)]thiourea and N-[2-(3-indolylethyl)]-N'-[2 (5-bromopyridyl)]thiourea were the most active agents and inhibited the LTC(4) release with low micromolar IC(50) values of 4.9 microM and 6.1 microM, respectively. The hydroxylphenyl substituted compounds N-[2-(4 hydroxyphenyl)ethyl]-N'-[2-(5-chloropyridyl)]thiourea (IC(50)=12.6 microM), N-[2 (4-hydroxyphenyl)ethyl]-N'-[2-(5-bromopyridyl)]thiourea (IC(50)=16.8 microM) and N-[2-(4-hydroxyphenyl)ethyl]-N'-[2-(pyridyl)]thiourea (IC(50)=8.5 microM) were the most active pyridyl thiourea agents. Notably, the introduction of electron withdrawing or donating groups had a marked impact on the biological activity of these thiourea derivatives and the Hammett sigma values of their substituents were identified as predictors of their potency. In contrast, experimentally determined partition coefficient values did not correlate with the biological activity of the thiourea compounds which demonstrates that their liphophilicity is not an important factor controlling their mast cell inhibitory effects. PMID- 12565957 TI - Discovery, SAR, synthesis, pharmacokinetic and biochemical characterization of A 192411: a novel fungicidal lipopeptide-(I). AB - The echinocandin class of cyclic lipopeptides has been simplified to discover potent antifungal compounds. Namely A-192411 shows good in vitro activity against common pathogenic yeasts and has an acceptable safety window in vivo. Discovery, limited SAR, synthesis, biochemical and pharmaco-dynamic profiles of A-192411 are presented. PMID- 12565958 TI - In vivo characterization of A-192411: a novel fungicidal lipopeptide (II). AB - The ability of the novel antifungal cyclic hexalipopetide A-192411 to treat fungal infections in rodents is presented. Efficacy was demonstrated against Candida albicans as both prolonged survival of systemically infected mice and clearance of viable yeasts from kidneys. The efficacy of A-192411, administered intraperitoneally, was equivalent to amphotercin B at a 4-fold lower dose by weight in the systemic candidiasis models in mice, while the efficacy of A-192441 administered intravenously was equivalent to amphotercin B by weight in the Candida pyelonephritis model in rats. A-192411 also slightly prolonged the survival of immunocompromised mice infected systemically with Aspergillus fumigatus. PMID- 12565959 TI - 1,4-Diazepane-2-ones as novel inhibitors of LFA-1. AB - The design, synthesis, and biological evaluation of 1,4-diazepane-2-ones as novel LFA-1 antagonists from a scaffold-based combinatorial library are described. Initial optimization of the library lead has resulted in high-affinity antagonists of the LFA-1/ICAM-1 interaction, such as compounds 18d and 18e with IC(50) values of 110 and 70 nM, respectively. PMID- 12565960 TI - Thiophene-based vitronectin receptor antagonists. AB - A series of alpha(v)beta(3) antagonists based on a thiophene scaffold were synthesized via two routes and evaluated for in vitro biological activity. We have identified several structurally similar antagonists with different selectivities towards alpha(IIb)beta(3), alpha(v)beta(5) and alpha(5)beta(1) at the cellular level. In addition, these antagonists exerted an antiangiogenic effect in the chick chorioallantoic membrane (CAM) assay. PMID- 12565961 TI - Structure-activity relationships of substituted benzothiophene-anthranilamide factor Xa inhibitors. AB - Compound 1 was identified by high throughput screening as a novel, potent, non amidine factor Xa inhibitor with good selectivity against thrombin and trypsin. A series of modifications of the three aromatic groups of 1 was investigated. Substitution of chlorine or bromine for fluorine on the aniline ring led to the discovery of subnanomolar factor Xa inhibitors. Positions on the anthranilic acid ring that can accommodate further substitution were also identified. PMID- 12565962 TI - Pharmacophore-based discovery of substituted pyridines as novel dopamine transporter inhibitors. AB - Abnormal dopamine signaling in brain has been implicated in several conditions such as cocaine abuse, Parkinson's disease and depression. Potent and selective dopamine transporter inhibitors may be useful as pharmacological tools and therapeutic agents. Simple substituted pyridines were discovered as novel dopamine transporter (DAT) inhibitors through pharmacophore-based 3D-database search. The most potent compound 18 has a K(i) value of 79 nM in inhibition of WIN35,248 binding to dopamine transporter and 255 nM in inhibition of dopamine reuptake, respectively, as potent as cocaine. Preliminary structure-activity relationship studies show that the geometry and the nature of the substituents on the pyridine ring determine the inhibitory activity and selectivity toward the three monoamine transporters. The substituted pyridines described herein represent a class of novel DAT inhibitors with simple chemical structures and their discovery provides additional insights into the binding site of DAT. PMID- 12565963 TI - Sordaricin antifungal agents. AB - Compounds based on sordaricin were prepared via organometallic addition onto a fully protected sordaricin aldehyde. The fungal growth inhibition profiles for these compounds were established and the results are presented here. The synthesis of homologated sordaricin as well as ether and ester derivatives is presented, and structural rearrangement products upon oxidation. These compounds were evaluated as agents to inhibit fungal growth. PMID- 12565964 TI - 5-Azidoepibatidine: an exceptionally potent photoaffinity ligand for neuronal alpha 4 beta 2 and alpha 7 nicotinic acetylcholine receptors. AB - Racemic 5-azidoepibatidine [(+/-)-1] was synthesized via 5-aminoepibatidine as a candidate photoaffinity ligand with exceptionally high affinity at the mammalian neuronal nicotinic receptors (K(i) values of 0.027 nM for alpha 4 beta 2 and 9.7 nM for alpha 7) and excellent photoreactivity. PMID- 12565965 TI - Synthesis, opioid receptor binding, and functional activity of 5'-substituted 17 cyclopropylmethylpyrido[2',3':6,7]morphinans. AB - A series of naltrexone-derived pyridomorphinans possessing various substituents at the 5'-position on the pyridine ring were synthesized and evaluated for opioid receptor binding in rodent brain membranes and functional activity in smooth muscle preparations. While the introduction of aromatic 1-pyrrolyl group (6h) improved the delta affinity and delta antagonist potency of the parent compound (3), the introduction of guanidine group (6i) transformed it to a kappa selective ligand in opioid receptor binding and [35S]GTP-gamma-S functional assays. PMID- 12565966 TI - Both 5-arylidene-2-thioxodihydropyrimidine-4,6(1H,5H)-diones and 3-thioxo-2,3 dihydro-1H-imidazo[1,5-a]indol-1-ones are light-dependent tumor necrosis factor alpha antagonists. AB - Based on the realization that N-alkyl 5-arylidene-2-thioxo-1,3-thiazolidin-4-ones are tumor necrosis factor-alpha antagonists, we discovered two additional classes of antagonists: 3-thioxo-2,3-dihydro-1H-imidazo[1,5-a]indol-1-ones (via rational design) and 5-arylidene-2-thioxodihydropyrimidine-4,6(1H,5H)-diones (via computer guided screening). Chemical modification of the lead structures showed that the structure-activity relationship profiles for both of these series were dependent on the electronic properties of the molecules. Subsequent studies showed that they were light-dependent inhibitors. PMID- 12565967 TI - Cephalosporin prodrugs of paclitaxel for immunologically specific activation by L 49-sFv-beta-lactamase fusion protein. AB - Paclitaxel conjugates of 7-phenylacetamidocephalosporanic acid were prepared as prodrugs for site specific activation by targeted beta-lactamase. Immunologically specific activation of the prodrug 5 containing 3,3-dimethyl-4-amino-butyric acid as linker in combination with the fusion protein L-49-sFv-beta-lactamase was demonstrated in vitro on 3677 melanoma cells. PMID- 12565968 TI - Novel inhibitors of IMPDH: a highly potent and selective quinolone-based series. AB - A series of novel quinolone-based small molecule inhibitors of inosine monophosphate dehydrogenase (IMPDH) was explored. The synthesis and the structure activity relationships (SARs) derived from in vitro studies are described. PMID- 12565969 TI - Quinolone-based IMPDH inhibitors: introduction of basic residues on ring D and SAR of the corresponding mono, di and benzofused analogues. AB - The synthesis and the structure-activity relationships (SAR) of analogues derived from the introduction of basic residues on ring D of quinolone-based inhibitors of IMPDH are described. This led to the identification of compound 27 as a potent inhibitor of IMPDH with significantly improved aqueous solubility over the lead compound 1. PMID- 12565970 TI - Synthesis and dopamine transporter affinity of chiral 1-[2-[bis(4 fluorophenyl)methoxy]ethyl]-4-(2-hydroxypropyl)piperazines as potential cocaine abuse therapeutic agents. AB - A series of optically pure phenyl-and non-phenyl-substituted 1-[2-[bis(4 fluorophenyl)methoxy]ethyl]-4-(2-hydroxypropyl)piperazines was synthesized and their binding affinity for dopamine transporter (DAT) was investigated. The analogues with a hydroxyl group in the S configuration were more selective for the DAT over the serotonin transporter (SERT) than the corresponding R enantiomers. Compound (+)-11 showed high affinity and selectivity for DAT over the SERT and, therefore, is a potential candidate for the development of a long acting cocaine abuse therapeutic agent. PMID- 12565971 TI - A facile synthesis of C(2)-symmetric 17 beta-estradiol dimers. AB - A rapid and efficient synthesis of a series of C(2)-symmetric 17 beta-estradiol dimers is described. The new molecules are linked at position 17 alpha of the steroid nucleus with either an alkyl chain or a polyethylene glycol chain. They are made from estrone in five chemical steps with an overall yield exceeding 30%. The biological activity of these compounds was evaluated in vitro on estrogen dependent and independent (ER(+) and ER(-)) human breast tumor cell lines: MCF-7 and MDA-MB-231. Some of the dimers present selective cytotoxic activity against the ER(+) cell line. PMID- 12565972 TI - Design, synthesis and structure-activity relationships of benzoxazinone-based factor Xa inhibitors. AB - A series of benzoxazinone derivatives was designed and synthesized as factor Xa inhibitors. We demonstrated that the naphthyl moiety in the aniline-based compounds 1 and 2 can be replaced with benzene-fused heterobicycles and biaryls to give factor Xa inhibitors with improved trypsin selectivity. The P4 modifications lead to monoamidines which are moderately active. The benzoxazinones 41-45 are potent against factor Xa, retain the improved trypsin selectivity of the corresponding aniline-based compounds, and show strong antithrombotic effect dose responsively. PMID- 12565973 TI - Piperazine-based CCR5 antagonists as HIV-1 inhibitors. III: synthesis, antiviral and pharmacokinetic profiles of symmetrical heteroaryl carboxamides. AB - The unsymmetrical nicotinamide-N-oxide moiety in compound 1 was replaced with symmetrical isonicotinamides as well as 4,6-dimethyl pyrimidine-5-carboxamides. Compound 16 from the latter set reduced the number of rotamers, improved potency of inhibiting UIV entry, slightly diminished the affinity for the muscarine receptors and showed very good oral absorption. PMID- 12565974 TI - Condensed aromatic peptide family of microbial metabolites, inhibitors of CD28 CD80 interactions. AB - Three condensed aromatic peptides SCH79235 (1), SCH79236 (2), and SCH204698 (3) were isolated from the fermentation broth of a Streptomycete microorganism. The structure of SCH204698 (3) was established by extensive NMR spectral data. All these compounds exhibited good activity against CD28-CD80 binding with an IC(50) of 0.42, 0.38 and 0.22 microM, respectively. PMID- 12565975 TI - Selective phenolic acylation of 10-hydroxycamptothecin using poly (ethylene glycol) carboxylic acid. AB - Selective acylation of the phenolic hydroxyl group of 10-hydroxycamptothecin has been accomplished using phenyl dichlorophosphate. Additional modification of the 10-OH as an ether permits a 20-acyl derivative to be synthesized. This result along with data from a 6-hydroxyquinoline model strongly suggests that powerful intermolecular hydrogen bonding exists in the parent molecule. PMID- 12565976 TI - Organic phenyl arsonic acid compounds with potent antileukemic activity. AB - A series of 12 organic arsonic acid compounds has been synthesized and evaluated against human B-lineage (NALM-6) and T-lineage (MOLT-3) acute lymphoblastic leukemia (ALL) cell lines. The lead compounds 2-trichloromethyl-4-[4'-(4" phenylazo)phenylarsonic acid]aminoquinazoline (compound 19, PHI-P518; IC(50)=1.1+/-0.5 microM against NALM-6 and 2.0+/-0.8 microM against MOLT-3) and 2 methylthio-4-(2'-phenylarsonic acid)aminopyrimidine (compound 15, PHI-P381; IC(50)=1.5+/-0.3 microM against NALM-6 and 2.3+/-0.5 microM against MOLT-3) exhibited potent antileukemic activity at low micromolar concentrations. PMID- 12565977 TI - The challenges and achievements involved in implementing Quality of Life research in cancer clinical trials. AB - Over the last decade, Quality of Life (QOL) research has become an important aspect of cancer clinical trials. A dramatically increasing number of published studies, both randomised and non-randomised, report QOL outcomes. There is increasing evidence that QOL results impact on both future research and treatment decisions for clinicians. The rising number of studies with QOL components is mirrored within the European Organization for Research and Treatment of Cancer (EORTC), one of the largest cancer clinical trial organisations in Europe. Clinical trial groups have frequently reported on the difficulties and challenges of implementing QOL research. In the following paper, we therefore examine past experience in EORTC QOL studies, with a focus on the challenges presented and the improved approaches that are being implemented to obtain more meaningful outcomes. PMID- 12565978 TI - Management of primary breast cancer in the elderly patient. PMID- 12565979 TI - Quality of life in patients with advanced colorectal cancer: what has been learnt? AB - Accurate assessment of health-related quality of life (HRQoL) in patients with advanced colorectal cancer is essential to improve our understanding of how cancer and chemotherapy influence patients' life and to adapt treatment strategies. Specific questionnaires have descriptive and predictive value and can be used to evaluate new therapies. Results from HRQoL assessments in randomised trials help patients and physicians to choose between treatment options. More than half of the patients treated with palliative chemotherapy have an improvement or at least a preservation of their HRQoL. However, several trials have found small differences in HRQoL between treatment groups. This may be due to the insufficient sensitivity of tools, low numbers of patients or missing data. An international consensus on the methods of measurement of HRQoL in oncology is warranted to enhance compliance, to better interpret results and to optimise the publication of precise HRQoL data. PMID- 12565980 TI - Health effects of the Chernobyl accident: fears, rumours and the truth. AB - The impact of the world's worst nuclear disaster at Chernobyl in 1986 is reviewed within a framework of a triad of fear, rumour and truth. The scope of the accident, Soviet secrecy about it, and the lack of general awareness of, or disregard for, the effects of radiation created a fertile ground for persistent fears and rumours attributing any health problem to Chernobyl. Scientifically correct answers to health issues have been the means to combat disinformation, and to replace interconnected fears, misconceptions and rumours. To date, according to the United Nations Scientific Committee on the Effects of Atomic Radiation (UNSCEAR) 2000 Report, based on a review of epidemiological and radiobiological studies, the main radiation-related effect of the Chernobyl accident is an increased risk of childhood thyroid cancer. In addition, the accident has had serious non-radiation-related psychological consequences on the residents of the contaminated territories, resettled populations and clean-up workers. Researchers in search of the truth through epidemiological reasoning are facing serious challenges which are reviewed within this article. PMID- 12565981 TI - Treatment of operable breast cancer in the elderly: a randomised clinical trial EORTC 10850 comparing modified radical mastectomy with tumorectomy plus tamoxifen. AB - We have examined the outcome of older patients with operable breast cancer treated in a randomised trial by either standard surgery or less extensive surgery and tamoxifen. There were 236 participants aged >/=70 years, randomised to either modified radical mastectomy MRM (n=120) or wide local excision (WLE) and tamoxifen (T) 20 mg daily (n=116). Survival curves were estimated using the Kaplan-Meier method and multivariate analyses were performed using Cox's proportional hazards model. Endpoints were survival and time to first relapse or progression, loco-regional progression, time to distant progression and progression-free survival. No significant difference was seen in terms of progression-free survival, but there were significantly more loco-regional relapses in the WLE+T group. In contrast, there were more distant metastases in the MRM group, but with a similar overall survival in both groups. The results of this trial give cautious support for the use of WLE+T for selected older women. PMID- 12565983 TI - Phase II study of LU 103793 (dolastatin analogue) in patients with metastatic breast cancer. AB - LU 103793 is a synthetic analogue of Dolastatin 15 that inhibits tubulin polymerisation. The aim of this study was to evaluate the efficacy and tolerability of LU 103793 in patients with metastatic breast cancer who had been previously treated with two lines of chemotherapy for advanced disease. Patients received LU 103793 at a dose of 2.5 mg/m(2)/day over 5 min for 5 consecutive days every 3 weeks. Thirty-four patients were enrolled and 23 patients were eligible for the evaluation of efficacy. Eleven patients experienced grade 4 neutropenia. Other related grade 3/4 adverse events included asthenia (three patients), stomatitis (1), myalgia (1) and increase of serum bilirubin (2). The main toxicity was hypertension occurring in seven out of 34 patients. There were no objective responses, 7 patients had stable disease. These results do not support the further evaluation of LU 103793 in metastatic breast cancer patients using this dose and schedule. PMID- 12565982 TI - Treatment of operable breast cancer in the elderly: a randomised clinical trial EORTC 10851 comparing tamoxifen alone with modified radical mastectomy. AB - For treatment of early breast cancer in older women, little evidence is available from randomised trials. We conducted a randomised trial comparing modified radical mastectomy (MRM) with tamoxifen (TAM) as the sole initial therapy in 164 patients aged >/=70 years with operable breast cancer. 82 were treated by MRM and 82 with TAM. Survival curves were estimated using the Kaplan-Meier method: multivariate analyses were performed using the Cox's proportional hazards model. Endpoints included survival, time to first relapse or progression, loco-regional progression, time to distant progression and progression-free survival. After a median follow-up of approximately 10 years, there was a significantly decreased time to progression in the TAM only group (logrank P<0.0001) and significantly shorter time to local progression within the TAM group (logrank P<0.0001). Overall survival of the two groups was similar. The results indicate that tamoxifen alone leads to an unacceptably high rate of local progression or relapse. PMID- 12565984 TI - Predictive factors for blastoid transformation in the common variant of mantle cell lymphoma. AB - Approximately 20% of the mantle cell lymphoma (MCL) patients present with the blastoid variant at diagnosis. Blastoid changes may occur also during the course of the disease, but factors related to blastoid transformation are poorly understood. In the present study, the incidence and predictive factors for blastoid transformation were analysed among 52 patients who primarily had the common variant of MCL and one or more biopsies taken at the time of disease progression. Blastoid transformation occurred in 18 (35%) patients. The minimum estimated risk of transformation was 42% at 5 years of follow-up. At the time of transformation, all except two patients had systemic lymphoma with lymphatic blasts in the blood. The median survival time after blastoid transformation was 3.8 months compared with 26 months in patients without transformation (P<0.001). The respective survival times as calculated from the initial diagnosis of MCL were 31 and 60 months. Leucocytosis, an elevated serum lactate dehyrdogenase (LDH) level, and a high proliferative activity at diagnosis as assessed by the mitotic count and Ki-67 staining were associated with an increased risk of blastoid transformation, and elevated serum LDH and blood leucocytosis with a short time interval to transformation. We conclude that blastoid transformation is not uncommon during the course of MCL, and is associated with a poor outcome. An elevated serum LDH level, a high cell proliferation rate, and leucocytosis are predictive for a high risk of blastoid transformation in MCL. PMID- 12565985 TI - Phase II study of XR 5000 (DACA), an inhibitor of topoisomerase I and II, administered as a 120-h infusion in patients with non-small cell lung cancer. AB - XR5000 is a tricyclic carboxamide-based cytotoxic agent that binds to DNA by intercalation and stimulates DNA cleavage by inhibition of both topoisomerase I and II. The aim of this study was to evaluate the antitumoral activity and safety profile of XR5000 given as second-line chemotherapy in patients with advanced non small cell lung cancer (NSCLC). Patients received XR5000 at the dose of 3010 mg/m(2) as a 120-h central venous infusion every 3 weeks. The 15 patients (median age 56 years, range 48-71 years) enrolled had an Eastern Cooperative Oncology Group (ECOG) performance status of 0 (3 patients), 1 (11 patients) or 2 (1 patient). A total of 32 cycles of XR5000 (median 2, range 1-6) were given to 14 patients. No objective response (assessed according to World Health Organization (WHO) criteria) was documented in the 12 evaluable patients by an external review panel; in 4 out of the 12 patients disease stabilisation was recorded. The following toxicities graded according to the Common Toxicity Criteria (CTC) version 2.0. were observed: one grade 3 and two grade 4 granulocytopenia, one grade 3 and one grade 4 thrombocytopenia, one grade 3 deep venous thrombosis, one grade 3 fatigue, and grade 3 undocumented epileptic seizures which led to death in 2 patients. With only 4 out of 12 patients reaching stable disease when using this dose and regimen, further evaluation of XR5000 in advanced NSCLC is not justified. PMID- 12565986 TI - Assessing the clinical significance of health-related quality of life (HrQOL) improvements in anaemic cancer patients receiving epoetin alfa. AB - Health-related quality of life (HrQOL) assessments are gaining importance as outcome measures in cancer clinical trials. A recently published clinical trial reported statistically significant (P<0.001) increases in haemoglobin (Hb) levels and significantly (P<0.01) increased HrQOL scores following the administration of recombinant human erythropoietin (r-HuEPO, epoetin alfa) versus placebo to anaemic cancer patients who received non-platinum chemotherapy. This study employed five cancer-specific HrQOL instruments. Hb and HrQOL data from this trial were analysed to estimate the minimally important difference (MID) in HrQOL measures that could be interpreted as clinically meaningful, with Hb level selected as the best external standard. Patients were assigned to two groups: improved (Hb increases of >/=1 g/dL) or stable (change in Hb of-1 g/dL to <1 g/dL). The MID was first determined as the difference between the mean changes in HrQOL in the improved group versus the stable group. By this analysis, the differences in HrQOL scores between the epoetin alfa group and the placebo group were clinically important for all Hb-sensitive, cancer-specific HrQOL evaluations. Linear regression analyses performed to provide estimates of the MID for specific values of Hb change confirmed that the differences in HrQOL scores between patient groups were clinically significant. These analyses were repeated using a data set from a separate clinical trial, which further supported the conclusion that observed HrQOL changes demonstrated in the multicentre, double blind study were clinically important. These methods provide one means for interpreting the clinical relevance of changes in HrQOL evaluated in clinical trials. PMID- 12565987 TI - High-dose 5-fluorouracil plus low dose methotrexate plus or minus low-dose PALA in advanced colorectal cancer: a randomised phase II-III trial of the EORTC Gastrointestinal Group. AB - The aim of this study was to investigate whether N-(phosphonacetyl)-L-aspartic acid (PALA) can enhance the activity of low-dose methotrexate (LD-MTX) modulated infusional 5-fluorouracil (5-FU) in patients with advanced colorectal cancer. 198 patients were randomised either to (i) 5-FU 60 mg/kg as a continuous infusion over 48 h, to be given weekly four times and thereafter every 2 weeks, with methotrexate 40 mg/m(2) administered just before 5-FU (control regimen) or to (ii) PALA 250 mg/m(2) as a 15 min infusion administered 24 h before 5-FU and methotrexate which was given as described in the control regimen. The response rate was 13% in the patients randomised to the control arm and 7% in the patients randomised to the experimental arm. If stabilisation of the disease was also considered as a positive response, these figures become 54 and 46%, respectively. All these differences did not reach statistical significance. The median durations of progression-free survival (PFS) in the two treatment groups were not significantly different. The median duration of survival was 13.1 months in the control arm and 11.9 months in the experimental arm (P=0.31). No benefit was obtained by adding PALA to LD-MTX+infusional FU. Taking into account data from US trials, the modulating effect of PALA, although 'promising' in phase II, could not be substantiated in randomised studies. PMID- 12565989 TI - Intellectual outcome in children and adolescents with acute lymphoblastic leukaemia treated with chemotherapy alone: age- and sex-related differences. AB - One of the most relevant concerns in long-term survivors of paediatric acute lymphoblastic leukaemia (ALL) is the development of neuropsychological sequelae. The majority of the published studies report on patients treated with chemotherapy and prophylactic central nervous system (CNS) irradiation, little is known about the outcome of patients treated with chemotherapy-only regimens. Using the standardised clinical and neuropsychological instruments of the SPOG Late Effects Study, the intellectual performance of 132 paediatric ALL patients treated with chemotherapy only was compared to that of 100 control patients surviving from diverse non-CNS solid tumours. As a group, ALL and solid tumour survivors showed normal and comparable intellectual performances (mean global IQ 104.6 in both groups). The percentage of patients in the borderline range (global IQ between 70 and 85) was comparable and not higher as expected (10% cases and 13% controls, expected 16%). Only 2 (2%) of the former ALL and 1 (1%) of the solid tumour patients were in the range of mental retardation (global IQ<70). Former known risk factors described in children treated with prophylactic CNS irradiation, like a younger age at diagnosis of ALL and female gender, remained valid in chemotherapy-only treated patients. The abandonment of prophylactic CNS irradiation and its replacement by a more intensive systemic and intrathecal chemotherapy led to a reduction, but not the disappearance of late neuropsychological sequelae. PMID- 12565988 TI - The activity of raltitrexed (Tomudex) in malignant pleural mesothelioma: an EORTC phase II study (08992). AB - We investigated the activity and toxicity of raltitrexed (Tomudex) as a single agent treatment in patients with Malignant Pleural Mesothelioma (MPM) in a multicentre phase II European Organization for Research and Treatment of Cancer (EORTC) study. This study enrolled chemonaive patients with histologically confirmed measurable MPM. Raltitrexed was administered at the dose of 3 mg/m(2) intravenous (i.v.) bolus on an outpatient basis every 3 weeks. A maximum of eight cycles was planned in cases with an absence of progression or unacceptable toxicity. 24 patients received a total of 104 courses. 5 patients (20.8%, 95% confidence interval (CI) 7.1-42.2%) had a partial response (PR), which was confirmed by an independent radiology committee. Toxicity was mild, with diarrhoea, nausea, vomiting, fatigue and neutropenia as the major side-effects, but not exceeding grade 3 toxicity. We conclude that raltitrexed has activity as a single agent in the treatment of MPM, and that further studies with this drug in MPM are warranted. PMID- 12565990 TI - No evidence of involvement of germline BACH1 mutations in Finnish breast and ovarian cancer families. AB - Recently BACH1, a novel putative DNA helicase mapping to chromosome 17q22, was reported to interact specifically with BRCA1, and was suggested to be a candidate gene for predisposition to breast and ovarian cancers. Here, we screened 214 breast and ovarian cancer patients from 151 Finnish families for germline BACH1 mutations by utilising conformation-sensitive gel electrophoresis (CSGE) and genomic sequencing analysis. Four sequence alterations were observed in the exon regions of BACH1, three of which have been previously reported and were classified as polymorphisms. In 1 patient, a novel heterozygous 3101C>T variant was observed resulting in a proline to leucine substitution at codon 1034 (Pro1034Leu). This amino acid change occurs in the BRCA1 binding domain of the BACH1 protein. Although the 3101C>T transition was also found in one of the 304 control individuals with an unknown cancer status, it still remains possible that this alteration could represent a rare disease-related allele in the population. Functional assays are needed to resolve the biological significance of this novel BACH1 missense variant. Altogether, the available data suggest that germline mutations in BACH1 are extremely rare. PMID- 12565991 TI - Does use of alternative medicine predict survival from cancer? AB - This study examines the association between alternative medicines (AM) and cancer survival. A national multicentre study was carried out in Norway in December 1992 to assess the prevalence of AM use among cancer patients. One of the aims of this study was to assess the association between AM and long-time survival. In January 2001, survival data were obtained with a follow-up of 8 years for 515 cancer patients. A total of 112 (22%) assessable patients used AM. During the follow-up period, 350 patients died. Death rates were higher in AM users (79%) than in those who did not use AM (65%). In a Cox regression model adjusted for demographic, disease and treatment factors, the hazard ratio of death for any use of AM compared with no use was 1.30, (95% Confidence Interval (CI) 0.99, 1.70; P=0.056), suggesting that AM use may predict a shorter survival. Sensitivity analyses strengthened the negative association between AM use and survival. AM use had the most detrimental effect in patients with an ECOG (Eastern Cooperative Oncology Group) performance status (PS) of 0 (hazard ratio for use=2.32, 95% CI, 1.44, 3.74, P=0.001), when compared with an ECOG PS of 1 or higher. The use of AM seems to predict a shorter survival from cancer. The effect appears predominantly in patients with a good PS. PMID- 12565992 TI - Expression of uracil DNA glycosylase (UDG) does not affect cellular sensitivity to thymidylate synthase (TS) inhibition. AB - Uracil DNA glycosylase (UDG) is a base excision repair enzyme responsible for the removal of uracil present in DNA after cytosine deamination or misincorporation during replication. Inhibition of thymidylate synthase (TS), an important target for cancer chemotherapy, leads to deoxythymidine triphosphate (dTTP) pool depletion and elevation of deoxyuridine monophosphate (dUMP) pools which may also result in the accumulation of deoxyuridine triphosphate (dUTP). Large quantities of dUTP are believed to overwhelm the pyrophosphatase dUTPase, leading to misincorporation of uracil into DNA. Uracil is removed from DNA by uracil DNA glycosylase (UDG) resulting in an abasic site, but since the ratio dUTP:dTTP may remain high during continuing TS inhibition uracil can become re-incorporated into DNA causing a futile cycle eventually leading to DNA damage and cell death. This study has used isogenic cell lines differing in their expression of UDG to investigate the role of this enzyme in sensitivity to the specific TS inhibitors, ZD9331 and raltitrexed. The study showed that although increased expression and activity of UDG may lead to increased cell growth inhibition after TS inhibition over the first 24 h of treatment (measured using 3-(4,5-dimethyl (thiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT), probably due to increased damage to single-stranded DNA, the level of enzyme expression does not affect cell viability or cell death (measured using clonogenic assay, cell counting of attached/detached cells and cleavage of both poly ADP-ribose polymerase (PARP) and caspase 3). Increased expression and activity of UDG did not affect sensitivity to TS inhibition at later time points (up to 72 h treatment). Therefore UDG does not appear to play a major role in the response to TS inhibition, at least in the model used, and the results suggest that other determinants of response previously investigated, such as TS and dUTPase, may be more important for the response to TS inhibition. PMID- 12565993 TI - Reliability of quantitative reverse-transcriptase-PCR-based detection of tumour cells in the blood between different laboratories using a standardised protocol. AB - Differences in methods of reverse-transcriptase (RT)-polymerase chain reaction (PCR)-based detection of tumour cells in the blood gives rise to conflicting results, and standardisation is urgently needed. This pilot study aimed to assess the variation of RT-PCR-based detection of tumour cells in blood between four different laboratories using a commercially available kit with a standardised protocol. This kit allows comparison of results from different laboratories and facilitates the investigation of the influence of pre-analytical parameters. All laboratories analysed identical sets of blood samples spiked with tumour cells in a concentration range of 1-100 tumour cells/ml. To study at which level variation was introduced, three kinds of sample sets were generated in which (i) tumour cell RNA was spiked in the RNA of mononuclear cells (MNC), (ii) tumour cells were spiked in isolated MNC, and (iii) tumour cells were spiked in blood. Real-time quantitative RT-PCR was used to detect and quantify cytokeratin 20 (CK20) expression, which is indicative for the presence of epithelial tumour cells. All laboratories were able to detect CK20 expression in all spiked-RNA samples with limited variation in expression levels between laboratories. There was a positive correlation between the amount of spiked tumour cell RNA and CK20 expression level. RT-PCR analysis of spiked-MNC samples resulted in more variation in the CK20 expression levels between laboratories, however again all spiked samples were reported to be positive by all of the laboratories. The evaluation of spiked blood samples gave rise to considerable quantitative and qualitative variation between the laboratories. Our results underline the importance and need for standardisation and extended quality control studies in the field of pre analytics. PMID- 12565994 TI - Interferon-gamma upregulates MUC1 expression in haematopoietic and epithelial cancer cell lines, an effect associated with MUC1 mRNA induction. AB - Epithelial mucin-1 (MUC1) is an important target antigen that it is overexpressed in both epithelial and haematological cancers including multiple myeloma (MM) and some lymphomas and leukaemias. MUC1 has adhesive and immunosuppressive properties, which may promote cancer progression. These studies evaluated the effect of IFNs on MUC1 expression, since these agents are widely used in clinical cancer therapy. MUC1 and interferon (IFN) receptor expression were measured by radioligand binding. Changes in MUC1 mRNA levels in response to IFN-gamma were assessed by semi-quantitative reverse transcriptase-polymerase chain reaction (RT PCR). IFN-gamma was found to be a more potent inducer of MUC1 expression than IFN alpha. 125I-IFN binding studies indicated that both IFN receptors were expressed in most of the cell lines. With IFN-gamma treatment, there was upregulation of MUC1 mRNA. IFN-gamma has a more consistent and more potent effect upon MUC1 induction than IFN-alpha. The ability to upregulate MUC1 across a broad range of cancer types by a clinically available cytokine, IFN-gamma, has important implications for enhancing immunotherapeutic approaches targeting MUC1. PMID- 12565996 TI - High-speed cell sorting: fundamentals and recent advances. AB - Cell sorters have undergone dramatic technological improvements in recent years. Driven by the increased ability to differentiate between cell types, modern advances have yielded a new generation of cytometers, known as high-speed cell sorters. These instruments are capable of higher throughput than traditional sorters and can distinguish subtler differences between particles by measuring and processing more optical parameters in parallel. These advances have expanded their use to facilitate genomic and proteomic discovery, and as vehicles for many emerging cell-based therapies. High-speed cell sorting is becoming established as an essential research tool across a broad range of scientific fields and is poised to play a pivotal role in the latest therapeutic modalities. PMID- 12565997 TI - Plasmon resonant particles for biological detection. AB - Several recent advances in the optical observation, fabrication, and bioconjugation of nanometer-sized gold or silver colloids have produced a robust new class of label. These plasmon resonant particle (PRP) conjugates have several important advantages: they are ultra-bright, so the light scattered from the individual particles can be viewed using a simple optical microscope system with a white light illumination source; they do not photo-bleach; PRPs can be prepared that preferentially scatter light of a chosen color; and it is possible to prepare bioconjugated PRPs that are stable in solution. These properties, and the automation of PRP identification, discrimination, and counting, have enabled the development of ultrasensitive, multicolor, and multiplex applications in the life science field. PMID- 12565998 TI - Single-cell kinase assays: opening a window onto cell behavior. AB - Recent advances in analytical techniques have made the performance of biochemical assays on individual mammalian cells possible. Of particular interest is the ability to measure the activation of kinases, enzymes with critical roles in virtually every aspect of cell physiology. Single-cell kinase assays promise to deliver a newfound understanding of the molecular mechanisms responsible for cellular control and behavior by revealing the dynamic nature of signal transduction networks in living cells. A recent exciting development is the potential to perform assays of multiple kinases simultaneously in a single cell. PMID- 12565999 TI - Single-cell electroporation. AB - Electroporation is a widely used method for the introduction of polar and charged agents such as dyes, drugs, DNA, RNA, proteins, peptides, and amino acids into cells. Traditionally, electroporation is performed with large electrodes in a batch mode for treatment of a large number of cells in suspension. Recently, microelectrodes that can produce extremely localized electric fields, such as solid carbon fiber microelectrodes, electrolyte-filled capillaries and micropipettes as well as chip-based microfabricated electrode arrays, have proven useful to electroporate single cells and subcellular structures. Single-cell electroporation opens up a new window of opportunities in manipulating the genetic, metabolic, and synthetic contents of single targeted cells in tissue slices, cell cultures, in microfluidic channels or at specific loci on a chip based device. PMID- 12566000 TI - Advances in the analysis of single mitochondria. AB - Mitochondria are both morphologically and functionally diverse, and this variety is thought to have important biological ramifications. The development of methods to probe the properties of individual mitochondria is therefore of utmost importance. Recent advances have been made using in situ microscopy techniques and methods to investigate isolated mitochondria, including flow cytometry, capillary electrophoresis, patch-clamping and optical trapping. Such techniques have been used to study metabolism, mitochondrial calcium homeostasis, mitochondrial membrane potential, apoptosis, and other properties. PMID- 12566001 TI - Microfluidic devices for DNA sequencing: sample preparation and electrophoretic analysis. AB - Modern DNA sequencing 'factories' have revolutionized biology by completing the human genome sequence, but in the race to completion we are left with inefficient, cumbersome, and costly macroscale processes and supporting facilities. During the same period, microfabricated DNA sequencing, sample processing and analysis devices have advanced rapidly toward the goal of a 'sequencing lab-on-a-chip'. Integrated microfluidic processing dramatically reduces analysis time and reagent consumption, and eliminates costly and unreliable macroscale robotics and laboratory apparatus. A microfabricated device for high-throughput DNA sequencing that couples clone isolation, template amplification, Sanger extension, purification, and electrophoretic analysis in a single microfluidic circuit is now attainable. PMID- 12566002 TI - Advanced polymers for DNA separation. AB - Recent research to improve matrices for DNA separation has resulted in the development of advanced polymers for use in capillary electrophoresis and, more generally, for electrophoresis in microchannels. To date, the most commonly used matrix is linear polyacrylamide (LPA). Unfortunately, the high-molecular weight LPA solutions required for achieving good resolution lead to very viscous solutions. Moreover, the coating ability of LPA is very poor. For these reasons, many research groups have developed low-viscosity matrices, which make microchannel filling easier, and self-coating matrices, which are able to reduce efficiently the electro-osmotic flow and the interaction of DNA with the capillary wall. To this purpose, thermo-adjustable viscosity polymers represent a very clever and interesting class of matrices. PMID- 12566003 TI - The theory of DNA separation by capillary electrophoresis. AB - The Human Genome has been sequenced in large part owing to the invention of capillary electrophoresis. Although this technology has matured enough to allow such amazing achievements, the physical mechanisms at play during separation have yet to be completely understood and optimized. Recently, new separation regimes and new physical mechanisms have been investigated. The use of free-flow electrophoresis and new modes of pulsed-field electrophoresis have been suggested, while we have observed a shift towards single nucleotide polymorphism analysis and microchip technologies. A strong theoretical basis remains essential for the efficient development of new methods. PMID- 12566004 TI - Ultrasensitive protein-DNA binding assays. AB - Protein-DNA binding assays have been used in a variety of fields from fundamental studies regarding the binding process itself, to serving as probes for the detection, quantification and separation of target analytes. These assays have been used for the study of protein-DNA complex stoichiometry, the detection of DNA damage, and real-time separation of free and bound complexes by electrophoretic mobility. Synthetic DNA oligonucleotides, known as aptamers, have been increasingly used for affinity binding assays to proteins, as well as for separation studies and as biosensors. Recent advances have been made in protein DNA binding assays using capillary electrophoresis, laser-induced fluorescence, fluorescence polarization, molecular beacons, and affinity chromatography. PMID- 12566005 TI - Multiplexed protein profiling on antibody-based microarrays by rolling circle amplification. AB - Multiplexed immunoassays on antibody-based protein microarrays are an attractive solution for analyzing biological responses in normal and diseased states. Recently, the feasibility and utility of these assays has been established as concerns about specificity and sensitivity are being overcome by careful quality control and amplification technologies such as rolling circle amplification (RCA). RCA-amplified protein chips can now profile up to 150 proteins in various substrates including serum, plasma, and supernatants with high sensitivity, broad dynamic range and good reproducibility. Diagnostic utility of RCA-amplified protein chips has been shown for multiplexed allergen testing. When allied with multivariate statistical analysis, RCA protein chips have the potential to identify multiplexed biomarker classifiers for disease diagnosis and drug response. PMID- 12566006 TI - A sense of closeness: protein detection by proximity ligation. AB - Highly specific and sensitive procedures will be required to evaluate proteomes. Proximity ligation is a recently introduced mechanism for protein analysis. In this technique, the convergence of sets of protein-binding reagents on individual target molecules juxtaposes attached nucleic acid sequences. Through a ligation reaction a DNA reporter sequence is created, which can be amplified. The procedure thus encodes detected proteins as specific nucleic acid sequences in what may be viewed as a reverse translation reaction. PMID- 12566007 TI - Chemical proteomics and its application to drug discovery. AB - The completion of the human genome sequencing project has provided a flood of new information that is likely to change the way scientists approach the study of complex biological systems. A major challenge lies in translating this information into new and better ways to treat human disease. The multidisciplinary science of chemical proteomics can be used to distill this flood of new information. This approach makes use of synthetic small molecules that can be used to covalently modify a set of related enzymes and subsequently allow their purification and/or identification as valid drug targets. Furthermore, such methods enable rapid biochemical analysis and small-molecule screening of targets thereby accelerating the often difficult process of target validation and drug discovery. PMID- 12566008 TI - The separation of biomolecules using capillary electrochromatography. AB - The unique properties of capillary electrochromatography such as high performance, high selectivity, minimum consumption of both reagents and samples, and good compatibility with mass spectrometry make this technique an attractive one for the analysis of biomolecules including peptides, proteins, carbohydrates, nucleosides and nucleotides. Irreversible adsorption between the biomolecules and the charged packing surface leads to a lack of reproducibility and serious peak tailing, so various approaches have been taken to overcome this and to improve the technique for future challenges. PMID- 12566009 TI - Stable isotope-coded proteomic mass spectrometry. AB - Developing the ability to quantify changes in protein abundance between cells subjected to a variety of physiological and environmental conditions is an extremely active area of proteome research. Although advances in chromatography, mass spectrometry instrumentation, and bioinformatics have contributed to producing a viable method for comparative proteome-wide analyses, the highest precision of quantitation is based, in part, upon improved methods for chemical and metabolic stable isotope labeling of proteins and peptides. The ability to quantify differences in protein expression and post-translational modifications using stable isotope labeling has been achieved, but insights into the biochemical mechanisms that will contribute to the development of new biotechnologies have yet to be realized. PMID- 12566010 TI - Advances in quantitative proteomics via stable isotope tagging and mass spectrometry. AB - The high-throughput identification and accurate quantification of proteins are essential components of proteomic strategies for studying cellular functions and processes. Techniques that are largely based on stable isotope protein or peptide labeling and automated tandem mass spectrometry are increasingly being applied in quantitative proteomic studies. Over the past year, significant progress has been made toward improving and diversifying these technologies with respect to the methods for stable isotope labeling, process automation and data processing and analysis. Advances in stable isotope protein labeling and recent biological studies that used stable isotope based quantitative proteomics techniques are reviewed. PMID- 12566011 TI - Novel approaches to map protein interactions. AB - Although we now have the sequence of the human genome at hand, we face the challenge of assigning function to the identified genes. Genes usually ascribe their function through proteins, and the role of proteins is to interact with other molecules. Therefore, if we could map the interactions of proteins we would be able to understand protein function. The challenge of mapping protein interactions is vast and many novel approaches have recently been developed for this task using molecular biology, mass spectrometry and chemiproteomic techniques. PMID- 12566012 TI - Molecular interaction in capillary electrophoresis. AB - At the present time, affinity capillary electrophoresis is a well-established method for the measurement of binding constants involving 1:1 interactions. Many recent applications of this technique have illustrated its versatility. If affinity capillary electrophoresis is to be widely applied to the study of all types of interactions it is imperative that we consider that higher order interactions may be present. Applying proper equations or developing new models appropriate to the description of such systems is essential to obtaining reliable binding information. PMID- 12566014 TI - Corneal thickness measurements and visual function abnormalities in ocular hypertensive patients. AB - PURPOSE: It has been suggested that a considerable subset of patients currently classified as having ocular hypertension may have thicker than average corneas that result in an overestimation of the true intraocular pressure (IOP). As a consequence, ocular hypertension patients with greater corneal thickness may be at a lower risk for functional damage, such as that detected by short-wavelength automated perimetry (SWAP). The purpose of this study was to evaluate the frequency of SWAP deficits in ocular hypertension patients and to correlate these findings with corneal thickness measurements in the same patients. DESIGN: Cross sectional observational study. METHODS: Sixty-eight ocular hypertension patients with normal optic disks and 63 normal subjects were included in the study. All participants underwent standard automated perimetry (SAP), SWAP, and central corneal thickness measurements using ultrasound pachymeter. Central corneal thickness measurements in ocular hypertension patients with abnormal visual field test results were compared with central corneal thickness measurements in ocular hypertension patients with normal visual field results. In addition, central corneal thickness measurements in ocular hypertension patients were compared with central corneal thickness measurements in normal subjects. RESULTS: Sixteen of 68 patients with ocular hypertension (24%) demonstrated SWAP abnormalities, whereas four of 68 (6%) showed a deficit on SAP. The mean central corneal thickness in ocular hypertension patients with abnormal SWAP results was significantly lower than the mean central corneal thickness in ocular hypertension patients with normal SWAP results (545 +/- 25 microm vs 572 +/- 35 microm; P =.006). The mean central corneal thickness in the normal group was 557 +/- 33 microm. The mean central corneal thickness in ocular hypertension patients with normal SWAP results was significantly higher than in normal subjects (P =.02). There was no significant difference between mean central corneal thickness in normal subjects and in ocular hypertension patients with abnormal SWAP results (P =.19). CONCLUSIONS: The patients classified as having ocular hypertension but with visual field loss detected by SWAP had significantly lower central corneal thickness measurements than the ocular hypertension patients with normal visual field results. These results suggest that central corneal thickness should be taken into account when assessing risk for the development of glaucomatous damage among ocular hypertension patients. PMID- 12566015 TI - Timolol 0.5%/dorzolamide 2% fixed combination vs timolol maleate 0.5% and unoprostone 0.15% given twice daily to patients with primary open-angle glaucoma or ocular hypertension. AB - OBJECTIVE: To compare the efficacy and safety of timolol 0.5%/dorzolamide 2% fixed combination vs timolol maleate 0.5% and unoprostone 0.15% given twice daily. DESIGN: Prospective multicenter, randomized, double-masked, crossover comparison study. METHODS: Primary open-angle glaucoma or ocular hypertension patients were randomly assigned to one of the treatment groups for a 6-week treatment period and then crossed over to the opposite treatment. Diurnal curve testing was performed at 8:00 AM, 10:00 AM, 4:00 PM, 6:00 PM, and 8:00 PM at baseline and the end of each treatment period. The run-in medicine was timolol twice daily for 28 days. RESULTS: Thirty-two patients completed this trial. The baseline trough pressure was 24.3 +/- 3.0 mm Hg, and the diurnal curve was 23.4 +/- 3.2 mm Hg. For the fixed combination the treatment trough pressure was 20.8 +/- 4.1 mm Hg and the diurnal curve was 19.6 +/- 3.6 mm Hg, whereas timolol and unoprostone concomitant therapy showed a treatment trough pressure of 20.1 +/- 4.5 mm Hg and a diurnal pressure of 19.8 +/- 4.1 mm Hg. There was no significant difference between treatment groups at any time point, for the diurnal curve, or in the extended reduction from baseline. There was no difference between treatment groups regarding ocular and systemic unsolicited or solicited adverse events. Burning, stinging, and conjunctival hyperemia were the adverse events most noted. There were no serious adverse events during this trial. CONCLUSIONS: This study suggests that both timolol/dorzolamide 2% fixed combination and concomitant timolol maleate 0.5% and unoprostone 0.15% therapy provide similar efficacy and safety throughout the daytime diurnal curve. PMID- 12566016 TI - Reduced cerebrovascular blood flow velocities and vasoreactivity in open-angle glaucoma. AB - PURPOSE: To compare cerebral blood flow velocities between open-angle glaucoma (OAG) patients and controls, at baseline and during hyperoxia. DESIGN: Observational cohort study. METHODS: A prospective study was conducted in a single institution. Sixteen OAG patients and 15 normal subjects, matched for age, were enrolled. Patients and controls were studied at baseline, while breathing room air, and during 100% oxygen breathing. The eye with the more severe visual field defect was chosen in glaucoma patients, while in controls, the study eye was chosen randomly. Subjects with history of diabetes, cardiovascular, or respiratory disease were excluded. Measurements included brachial arterial pressure, heart rate, intraocular pressure and transcranial Doppler (TCD). Mean and peak velocity and pulsatility index of the ipsilateral middle cerebral artery (MCA) were measured by TCD. RESULTS: At baseline, MCA mean and peak systolic blood flow velocities were significantly lower in glaucoma patients compared with controls velocities were significantly lower in glaucoma patients compared with controls (mean velocity: 50.2 vs 65.3 cm/s, P <.05; peak velocity: 74.2 vs 96.8 cm/s, P <.05). Additionally, while hyperoxia significantly decreased both mean and peak systolic velocities in MCA of controls (mean velocity: 65.3 vs 57.7 cm/s, P <.05; peak velocity 96.8 vs 87.9 cm/s, P <.05), it did not cause any significant change in OAG patients. CONCLUSIONS: Glaucoma patients were found to have lower MCA blood flow velocities and an absence of vasoreactivity to hyperoxia, compared with controls. The relationship of these cerebral hemodynamic abnormalities to glaucoma pathogenesis and progression remains to be explored. PMID- 12566017 TI - Structure and function evaluation (SAFE): II. Comparison of optic disk and visual field characteristics. AB - PURPOSE: To evaluate the relationship between glaucomatous structural damage to the optic nerve and development of visual field loss with standard automated perimetry (SAP) and short wavelength automated perimetry (SWAP). DESIGN: Cohort study. METHODS: Patients with elevated intraocular pressure and normal SAP visual fields were enrolled in this prospective study. Stereo optic disk photographs, SAP, and SWAP visual fields were obtained annually over a period of 4 or more years. Trained readers evaluated baseline and follow-up optic disk photographs for evidence of glaucomatous damage. Standard automated perimetry and SWAP examinations were evaluated according to previously validated criteria for development of confirmed visual field changes. RESULTS: Two-hundred ninety-five subjects (479 eyes) were enrolled. Following masked assessment of stereo photographs by an optic disk reading center, 272 of the 479 eyes were judged to have glaucomatous optic neuropathy at the time of study entry. Depending on the criteria employed, approximately 10% to 17.5% of all eyes developed confirmed visual field loss for SAP (conversions). Of the conversions, 75% to 80% had baseline glaucomatous optic disk damage, whereas normal and glaucomatous optic disks were equally divided (50%) among the nonconversion eyes. This difference was statistically significant (P <.003). Depending on the criteria employed, 4% to 12% of the eyes had confirmed SWAP deficits at baseline, and 4% to 8% developed confirmed SWAP defects at a follow-up examination. There was a greater percentage of eyes with a glaucomatous optic neuropathy in the group with SWAP deficits (75%-100%) than for those eyes in which SWAP remained normal (45%-60%). Some of these differences were statistically significant (P <.05). CONCLUSIONS: A strong relationship exists between glaucomatous optic disk damage at study entry and the subsequent development of a confirmed glaucomatous SAP visual field defect. A higher percentage of glaucomatous optic disks were also found in patients with SWAP deficits at baseline and in those who later developed SWAP deficits. These findings support the premise that a glaucomatous optic disk is predictive of the subsequent development of glaucomatous visual field loss. PMID- 12566018 TI - Use of glaucoma drainage devices in the management of glaucoma associated with aniridia. AB - PURPOSE: To describe intraocular pressure (IOP) and visual acuity outcomes after glaucoma drainage device placement in eyes with glaucoma associated with aniridia. DESIGN: Retrospective, noncomparative, consecutive, interventional case series. METHODS: The medical records of all patients who underwent glaucoma drainage device placement for the management of glaucoma associated with aniridia at the Bascom Palmer Eye Institute between January 1, 1989, and December 31, 1999, were reviewed. RESULTS: The study included eight eyes of five patients with a median age at the time of glaucoma drainage device placement of 92 months (range, 10- 495 months) and a median follow-up of 19 months (range, 11- 33 months). Preoperatively, the mean IOP was 35 mm Hg using a mean of one antiglaucoma medication. Postoperatively, the mean IOP was reduced to 14.9 mm Hg and no patient was using antiglaucoma medication. Final visual acuity was improved compared with preoperatively in five of eight eyes (63%) and remained unchanged in two eyes (25%). Loss of light perception occurred in one eye owing to retinal detachment. Success rates using Kaplan-Meier survival analysis were 100% at 6 months and 88% at 1 year. CONCLUSIONS: Glaucoma drainage device placement for glaucoma associated with aniridia achieves IOP control and vision preservation in most patients. PMID- 12566019 TI - Combined use of frequency doubling perimetry and polarimetric measurements of retinal nerve fiber layer in glaucoma detection. AB - PURPOSE: The aim of this study was to evaluate the diagnostic usefulness of the combined use of frequency-doubling technology (FDT) perimetry and polarimetry of the retinal nerve fiber layer. DESIGN: Cross-sectional study. METHODS: Seventy ocular hypertensive patients (normal optic disk and standard perimetry, elevated intraocular pressure [>21 mm Hg]), 59 patients with "preperimetric" open-angle glaucoma (glaucomatous optic disk atrophy, elevated intraocular pressure [>21 mm Hg], no visual field defect in standard perimetry), 105 patients with "perimetric" open-angle glaucoma (glaucomatous optic disk atrophy and clearly marked visual field defect), and 73 control subjects had FDT screening (protocol: C-20-5) and polarimetric measurements (GDx). Criteria for exclusion: optic disks larger than 4 mm(2), media opacities, patients younger than 33 years or older than 66 years. None of the subjects had earlier FDT perimetry. One eye of each patient and control subject entered the statistical evaluation. Database and statistical software were used for case-wise recalculation of all missed localized probability levels to create a FDT screening score. RESULTS: At a predefined specificity of 94.5% in control eyes, discrimination between "perimetric" glaucoma and normal subjects is superior using the FDT perimetry (sensitivity = 84.8%) in comparison to polarimetry (sensitivity = 63.8%), whereas sensitivity is similar with both methods in "preperimetric" patients (GDx, FDT: 25.4%). In several cases, patients classified as glaucomatous by the GDx are not the same patients as identified by the FDT perimetry. Therefore, a two dimensional discrimination analysis can increase correct positive classification. Using a linear combination of the present FDT screening score and polarimetry ("the number"), 92.4% of "perimetric" glaucoma eyes and 44.1% of "preperimetric" glaucoma eyes have been classified as glaucomatous. CONCLUSION: Joint usage of polarimetry and FDT perimetry indicate that a combination of different techniques which can uncover different glaucoma properties, might be helpful in early glaucoma detection. PMID- 12566020 TI - Optical coherence tomography for evaluating diabetic macular edema before and after vitrectomy. AB - PURPOSE: To report the use of optical coherence tomography (OCT) for evaluation of diffuse diabetic macular edema (DME) before and after vitrectomy. DESIGN: Interventional case series. METHODS: A retrospective study was made of 15 consecutive eyes of 13 patients that had vitrectomy for diffuse DME and OCT preoperatively and postoperatively. In seven eyes of six patients (group 1), vitrectomy was performed because of vitreomacular traction observed on biomicroscopy or OCT. In the other eight eyes of seven patients (group 2), vitrectomy was performed for DME not responsive to laser photocoagulation, with no vitreomacular traction on biomicroscopy or OCT. RESULTS: Mean +/- standard deviation (SD) follow-up after vitrectomy was 18 +/- 10 months (range, 6 to 33 months). In group 1, mean +/- SD retinal thickness decreased significantly from 661 +/- 181 microm preoperatively to 210 +/- 32 microm at the end of follow-up (P =.018). Median best-corrected visual acuity (BCVA) improved from 20/100 before surgery (range, 20/250 to 20/50) to 20/80 at the end of follow-up (range, 20/250 to 20/25; P =.046). In one eye in group 1, vitreomacular traction was only observed on OCT and not on biomicroscopy. In group 2, mean +/- SD retinal thickness decreased from 522 +/- 103 microm preoperatively to 428 +/- 121 microm at the end of follow-up (P =.2). Median BCVA was 20/100 before vitrectomy (range, 20/320 to 20/63) and 20/200 at the end of follow-up (range, 20/250 to 20/63; P =.78). CONCLUSIONS: Vitrectomy was beneficial in eyes with diffuse DME combined with vitreomacular traction but not in eyes without traction. Optical coherence tomography allowed diagnosis of subtle vitreomacular traction and provided precise preoperative and postoperative assessments of macular thickness. PMID- 12566021 TI - Iatrogenic punctate chorioretinopathy after internal limiting membrane peeling. AB - PURPOSE: To define the chorioretinal lesions created unintentionally during grasping the internal limiting membrane (ILM) with end-gripping forceps. DESIGN: Interventional case series. METHODS: A prospective review of a consecutive series of 15 patients (15 eyes) who underwent macular hole repair was performed. All patients underwent a three-port pars plana vitrectomy with ILM peeling. Fundus photographs, fluorescein angiography, and indocyanine green angiography were performed in all patients before and after surgery. RESULTS: In all eyes, small punctate barely seen chorioretinal lesions in the macular region were detected after surgery. These punctate lesions corresponded to the area where ILM was grasped with forceps. The lesions ranged from 100 to 400 microm in diameter. Their number ranged between 8 and 15 (mean, 12.6). In the early phase of fluorescein angiography, the lesions appeared hypofluorescent; in the late phase, there was slight staining of the margin of the lesions extending from the adjacent choriocapillaris, causing indistinct borders. In the early phase of indocyanine green angiogram, the lesions were hypofluorescent and in some larger lesions choroidal vessels were visible due to the thinning and atrophy of the retinal pigment epithelium and choriocapillaris. In the late phase, the homogenous hypofluorescent areas had well-delineated margins and were usually round or oval. This angiographic finding was independent of the age of the lesion, and neither enlargement of the lesions nor development of choroidal neovascularization were noted during follow-up. CONCLUSIONS: Iatrogenic punctate chorioretinopathy is the chorioretinal lesion created unintentionally during grasping the ILM with end-gripping forceps. It must be reported as one of the complications of ILM peeling. These small punctate lesions did not appear to affect the surgical outcome. However, the lesions should be followed to detect any long-term complication such as choroidal neovascularization. PMID- 12566022 TI - Retinal detachment rate after vitrectomy for retained lens material after phacoemulsification. AB - PURPOSE: To report the rate of retinal detachment after vitrectomy for retained intravitreal lens material after phacoemulsification using specific vitrectomy techniques designed to minimize retinal detachment. DESIGN: Consecutive, interventional case series. METHODS: Retrospective chart review of 100 consecutive eyes (one surgeon) of 100 patients undergoing vitrectomy for retained lens material after phacoemulsification and followed up for 3 months or longer unless an outcome event had occurred. Vitrectomy techniques employed to minimize the frequency of retinal detachment included inducing posterior vitreous detachment with maximal vitreous removal before phacofragmentation to avoid vitreous trauma, lens fragment debulking before fragmentation, use of low energy with high aspiration during removal of retained lens material, and intraoperative indirect ophthalmoscopic evaluation of the retinal periphery with scleral indentation to diagnose and treat intraoperative retinal breaks. The main outcome measures included prevalence of coexisting retinal detachment in eyes with retained lens material, incidence of retinal detachment or retinal breaks after vitrectomy for removal of retained lens material, and final visual acuity. RESULTS: The prevalence of previtrectomy retinal detachment was 4%; the incidence of postvitrectomy retinal detachment was 4%; the final visual acuity was 20/40 or better in 53%. One patient had a retinal break recognized during vitrectomy and was treated with retinocryopexy, but postoperative retinal detachment developed from a separate break. Three others were treated during vitrectomy for retinal breaks (including two with known preexisting breaks) and did not have any retinal detachment. Poor previtrectomy visual acuity (hand motions) was a risk factor for postvitrectomy retinal detachment. CONCLUSIONS: The rate of retinal detachment reported after vitrectomy for retained lens material after phacoemulsification can be minimized to approximately the rate expected with cataract extraction complicated by vitreous loss by employing standard surgical techniques. Higher risk eyes may benefit from more frequent postvitrectomy examinations. PMID- 12566023 TI - Vitreous opacities and outcome of vitreous surgery in patients with familial amyloidotic polyneuropathy. AB - PURPOSE: To report the prevalence of vitreous opacities and the outcome of vitreous surgery in patients with familial amyloidotic polyneuropathy (FAP). DESIGN: Observational case series. METHODS: In 37 patients with FAP and the ATTR Val30 Met mutation, vitreous opacities were present in 14 eyes of 9 patients and vitrectomy combined with phacoemulsification and intraocular lens implantation was performed in five eyes of three patients. In six patients with the ATTR Tyr114Cys mutation, vitreous opacities were present in both eyes of all six patients and vitrectomy combined with phacoemulsification and intraocular lens implantation was performed in nine eyes of six patients. The mean follow-up period after vitreous surgery was 20.9 +/- 16.8 months (range, 3 to 52 months). RESULTS: The prevalence of vitreous opacities is much higher in patients with ATTR Tyr114Cys (100%) than in those with ATTR Val30 Met (24%). The mean age at the onset of vitreous opacities was significantly lower in the patients with ATTR Tyr114Cys (37.0 +/- 5.3 years) than in the nine patients with ATTR Val30 Met (52.8 +/- 9.1 years; P <.005). Visual acuity improved in all 14 eyes after vitreous surgery; however, final visual acuity decreased in one eye owing to the occurrence of a central retinal vein occlusion. Vitreous opacities mildly increased in two eyes. CONCLUSIONS: Our data suggest that the ATTR Val30 Met and ATTR Tyr114Cys mutations induce different clinical features of vitreous opacities. Vitreous surgery combined with phacoemulsification and implantation of an intraocular lens is a safe and useful treatment. Careful long-term follow-up should be performed. PMID- 12566025 TI - Visual function index (VF-14) in exudative age-related macular degeneration of long duration. AB - PURPOSE: To evaluate the Visual Function Index (VF-14) questionnaire for its effectiveness in assessing visual function in patients with longstanding exudative age-related macular degeneration (AMD). DESIGN: Observational case series. METHODS: The records of 167 consecutive patients with recent neovascularization related to AMD between June 1990 and December 1994 at the Helsinki University Eye Clinic were analyzed in 1999. Of 121 patients still living, 74 (61%) attended the reexamination. After exclusions, data from 62 patients were analyzed. The VF-14 score, plus global assessment scores of satisfaction with vision and quality of vision, in which patients graded the subjective level of difficulty with their vision, best-corrected visual acuity (BCVA), contrast sensitivity, the area of the AMD lesion, and the shortest distance and direction from the center of the fovea to the edge of the subfoveal lesion, were analyzed. RESULTS: The VF-14 score correlated significantly with BCVA (P <.01), contrast sensitivity (P <.01), and global assessment scores (P <.01), showing stronger correlations with global assessment scores than did BCVA. In multivariate regression analysis, the global assessment scale of overall quality of vision and BCVA in the better eye were significant predictors (P <.001) of the variability in the VF-14 score. CONCLUSIONS: The VF-14 reflects visual function of patients with late AMD more effectively than BCVA measurement alone. The VF-14 can thus be used to compare the visual handicap of late AMD patients with that of patients with other eye diseases. PMID- 12566024 TI - A computerized method of visual acuity testing: adaptation of the early treatment of diabetic retinopathy study testing protocol. AB - PURPOSE: To develop a computerized method of visual acuity testing for clinical research as an alternative to the standard Early Treatment for Diabetic Retinopathy Study (ETDRS) testing protocol, and to evaluate its test-retest reliability and concordance with standard ETDRS testing. DESIGN: Test-retest reliability study. METHODS: Multicenter setting of a study population of 265 patients at three clinical sites. Visual acuity was measured with both the electronic visual acuity testing algorithm (E-ETDRS) and standard ETDRS protocol (S-ETDRS) twice on one eye of each patient. E-ETDRS testing was conducted using the electronic visual acuity tester (EVA), which utilizes a programmed Palm (Palm, Inc, Santa Clara, California, USA) hand-held device communicating with a personal computer and 17-inch monitor at a test distance of 3 meters. RESULTS: For the E-ETDRS protocol, test-retest reliability was high (r = 0.99; with 89% and 98% of retests within 0.1 logMAR and 0.2 logMAR of initial tests, respectively) and comparable with that of S-ETDRS testing (r = 0.99; with 87% and 98% of retests within 0.1 logMAR and 0.2 logMAR of initial test, respectively). The E-ETDRS and S-ETDRS scores were highly correlated (r = 0.96 for initial tests and r = 0.97 for repeat tests). Based on estimates of 95% confidence intervals, a change in visual acuity of 0.2 logMAR (10 letters) from a baseline level is unlikely to be related to measurement variability using either the E-ETDRS or the S-ETDRS visual acuity testing protocol. CONCLUSIONS: The E-ETDRS protocol has high test-retest reliability and good concordance with S-ETDRS testing. The computerized method has advantages over the S-ETDRS testing in electronically capturing the data for each tested letter, requiring only a single distance for testing from 20/12 to 20/800, potentially reducing testing time, and potentially decreasing technician-related bias. PMID- 12566026 TI - A novel mutation in the RDS/Peripherin gene causes adult-onset foveomacular dystrophy. AB - PURPOSE: To describe a novel mutation in the RDS/Peripherin gene that results in a moderately severe form of adult-onset foveomacular dystrophy. DESIGN: Observational case series. METHODS: Selected members of a family with adult-onset foveomacular dystrophy underwent complete ophthalmic evaluation, including fundus photography and fluorescein angiography, in a tertiary care referral center. The study population consisted of 12 members of a Caucasian kindred. After providing informed consent, patients donated blood for genomic DNA extraction and mutational screening using standard techniques. The main outcome measure were the presence of a RDS/Peripherin gene mutation in a patient with the disease and its absence in unaffected family members and controls. RESULTS: Eight affected family members and no unaffected family members demonstrated a single guanine base deletion at nucleotide 112 that led to premature termination at amino acid 38 of RDS/Peripherin polypeptide. This frameshift mutation results in truncation of nearly 90% of the gene product, thus probably representing a null allele. That results in a relatively severe phenotype, with choroidal neovascularization developing in two patients and geographic atrophy involving the macula in three patients. CONCLUSIONS: We describe a frameshift null mutation in the RDS/Peripherin gene associated with a relatively severe manifestation of adult onset foveomacular dystrophy in affected family members. PMID- 12566027 TI - Scleritis and other ocular side effects associated with pamidronate disodium. AB - PURPOSE: To evaluate reported cases of scleritis and other ocular side effects associated with pamidronate disodium, with emphasis on previously unreported cases of scleritis. DESIGN: Observational case series. METHODS: Case reports from the National Registry of Drug-Induced Ocular Side Effects (Casey Eye Institute), the Food and Drug Administration, the World Health Organization, and the literature were reviewed to determine possible adverse ocular side effects associated with pamidronate disodium. The World Health Organization's Causality Assessment Guide was used to categorize an adverse drug reaction. RESULTS: Seventeen cases of unilateral scleritis and one case of bilateral scleritis occurred, usually within 6 hours to 2 days after intravenous pamidronate disodium. Six patients had positive rechallenge testing with the scleritis occurring after a repeat drug exposure. Other ocular side effects with positive rechallenge data, associated with pamidronate disodium, include blurred vision, nonspecific conjunctivitis, ocular pain, bilateral anterior uveitis, and episcleritis. CONCLUSIONS: This is the first report, with rechallenge data, of any drug causing scleritis. Pamidronate disodium can cause vision-threatening diseases, which may require discontinuing the drug in some uveitis cases and, in this series, all cases of scleritis. PMID- 12566028 TI - Epidemic and sporadic cases of nontuberculous mycobacterial keratitis associated with laser in situ keratomileusis. AB - PURPOSE: To report national case-finding results for nontuberculous mycobacterial keratitis and describe its association with laser in situ keratomileusis (LASIK). DESIGN: Enhanced passive disease reporting. METHODS: In April 2001, we investigated a California cluster of Mycobacterium chelonae keratitis associated with hyperopic LASIK using a contact lens mask. To identify other possibly related cases, the American Academy of Ophthalmology e-mailed its members asking them to report recent cases of nontuberculous mycobacterial keratitis to the Centers for Disease Control and Prevention. RESULTS: Forty-three additional cases of keratitis were reported (onsets between August 2000 and June 2001). Of these, 31 occurred as part of two unrelated LASIK-associated outbreaks. The 12 other reported cases occurred in sporadic fashion. Of the latter cases, 4 were associated with LASIK surgery. None of the reported cases were related to the M. chelonae cluster in California. CONCLUSIONS: Laser in situ keratomileusis associated keratitis with nontuberculous mycobacteria may be more common than previously known. PMID- 12566029 TI - Malignant glaucoma due to drug-related angioedema. AB - PURPOSE: To report postsurgery angioedema resulting in malignant glaucoma. Interventional case report. METHODS: Three hours after uncomplicated cataract surgery on the right eye, a 61-year-old woman developed angioedema with swelling of the parapharyngeal tissue. Visual acuity deteriorated, and tonometry revealed an intraocular pressure of 60 mm Hg, with shallow anterior chambers, in both eyes. RESULTS: Ultrasound showed choroidal effusion on both eyes. Intraocular pressure could only be controlled surgically by procedure to deepen the anterior chamber. The angioedema regressed after withdrawal of candesartan, an angiotensin II antagonist that the patient had taken for 1 year. CONCLUSIONS: Angioedema without urticaria is well documented in patients receiving angiotensine converting enzyme inhibitors or angiotensin II antagonists. Drug-related angioedema may lead to a choroidal effusion syndrome with malignant glaucoma. Surgical intervention may trigger angioedema. Most important in treatment is withdrawal from the implicated medication and control of intraocular pressure. PMID- 12566030 TI - Increased disk size in glaucomatous eyes vs normal eyes in the Reykjavik eye study. AB - PURPOSE: To evaluate the use of disk diameter as an indicator in the identification of glaucomatous optic neuropathy. METHODS: We evaluated all available stereofundus photographs for 1,040 right eyes obtained in the Reykjavik Eye Study. Horizontal and vertical disk diameters were determined in a masked manner by a glaucoma specialist (K.F.D.). All disk diameters were corrected for refractive error. RESULTS: There were significant differences (P <.05) between the corrected vertical disk diameters of normal subjects (0.189 +/- 0.018 inches) and those suspected of having glaucoma (0.202 +/- 0.020 inches) as well as between the normal and the glaucoma groups (0.206 +/- 0.029 inches). The corrected horizontal measurement showed the same pattern. CONCLUSIONS: In the Reykjavik Eye Study, optic disks meeting structural criteria for glaucoma are significantly larger than normal nerves. PMID- 12566031 TI - Iris abscess as an unusual presentation of endogenous endophthalmitis in a patient with bacterial endocarditis. AB - PURPOSE: To report the clinical findings and management of a case of endogenous endophthalmitis in a patient with bacterial endocarditis presenting with a septic metastasis to the iris. DESIGN: Observational case report. METHODS: Review of clinical findings and treatment. RESULTS: A 37-year-old intravenous drug user hospitalized with bacterial endocarditis secondary to methicillin-sensitive Staphylococcus aureus bacteremia presented with a painful red left eye, hypopyon, and iris abscess. Roth spots were noted in the fundus of the right eye. Aqueous culture was positive for methicillin-sensitive S aureus. The patient was treated with intravitreal, topical, and intravenous antibiotics. The hypopyon and iris abscess resolved within 2 weeks, and the patient achieved a final visual acuity of 20/25 in the left eye. CONCLUSIONS: Septic metastasis to the iris is a rare occurrence. To our knowledge this is the first reported case of an iris abscess secondary to bacterial endocarditis. PMID- 12566032 TI - Lenticular fibroxanthomatous nodule. AB - PURPOSE: To describe two patients with unique lenticular nodular proliferations. DESIGN: Observational case reports. METHODS: The clinical histories and pathologic findings of two patients with lenticular nodular proliferations were reviewed. RESULTS: One patient with persistent hyperplastic primary vitreous and another patient with trauma developed lenticular nodular proliferations. The nodules were vascularized collections of foamy histiocytes, multinucleated cells, lens capsule, and lens epithelium that had undergone fibrous metaplasia. The lesions were classified as lenticular fibroxanthomatous nodules. CONCLUSIONS: A lenticular fibroxanthomatous nodule is a unique clinicopathologic entity that should be differentiated from Soemmerring ring, Elschnig pearl, and other simulating entities such as juvenile xanthogranuloma. PMID- 12566033 TI - Cataract after glaucoma filtration surgery. AB - PURPOSE: We reviewed the evidence regarding cataract incidence and progression after glaucoma surgery. DESIGN: Evidence-based review of available literature. METHODS: We performed a systematic review of relevant studies published in English. RESULTS: Reports of visually significant cataract after glaucoma surgery are supported by data from randomized clinical trials and large case series. CONCLUSIONS: Patients undergoing glaucoma surgery are at increased risk for the development and progression of cataract. PMID- 12566034 TI - Recurrent retinal vein occlusion in a patient with increased plasma levels of histidine-rich glycoprotein. AB - PURPOSE: To report an association between retinal vein occlusion and increased plasma levels of histidine-rich glycoprotein. DESIGN: Observational case report. METHODS: A 56-year-old woman presented with sudden and painless decrease of visual acuity of her right eye. Indirect ophthalmoscopy revealed retinal vein occlusion. She had experienced central retinal vein occlusion in this eye 6 years earlier. RESULTS: The patient's medical history was negative for cardiovascular risk factors. Further investigation into possible causes revealed increased values for histidine-rich glycoprotein. CONCLUSIONS: When ordering laboratory tests in patients with retinal vein occlusion to rule out coagulation disorders, increased plasma levels of histidine-rich glycoprotein should be considered in the differential diagnosis. Selective screening may be helpful in identifying retinal vein occlusion patients with thrombophilic defects, especially in young individuals with recurrent retinal vein occlusion in the absence of recognized cardiovascular risk factors. PMID- 12566035 TI - Trypan blue not toxic for retinal pigment epithelium in vitro. AB - PURPOSE: To investigate whether trypan blue has a toxic effect on cultured retinal pigment epithelial (retinal pigment epithelium) cells. DESIGN: Experimental study with a direct live/dead cell staining technique using fluorescent dyes. METHODS: Cultured human retinal pigment epithelium cells were exposed for 5 minutes to various concentrations of trypan blue (0.06%, 0.15%, 0.30%), and cell viability was confocally measured. RESULTS: No increased cell death was found in cultures incubated in any of the trypan blue concentrations used. CONCLUSION: These findings indicate that a short exposure of trypan blue does not have a toxic effect on cultured retinal pigment epithelium cells. PMID- 12566036 TI - Small-size pediatric vitrectomy wide-angle contact lens. AB - PURPOSE: We describe a new wide-field vitrectomy contact lens specifically designed to facilitate pediatric vitreoretinal surgery. DESIGN: Experimental study, instrument description. METHODS: A wide-field contact two-piece glass vitrectomy lens was designed with the largest diameter of 13.6 mm and a contact diameter of 7.00 mm, sacrificing only the more tilt-sensitive parts of the optics in order to reduce its physical size. RESULTS: Laboratory measurements in a model eye demonstrated a 61-degree binocular field of view, a 94-degree static field of view, and a maximum 129-degree field of view. Image magnification was 0.50 x. No complications were linked to the use of this instrument, and the need to exchange for another lens was never encountered in 10 cases. The two-piece glass construction enabled this lens to be sterilized in an autoclave or sterilized by any other instrument sterilization technique. CONCLUSION: The instrument successfully fulfilled the purpose of its design, providing at the same time an excellent wide field of view and ample space for the surgeon, performing well in 10 pediatric vitreoretinal cases (8 cases of retinopathy of prematurity; 2 cases of traumatic eye injury). Its loose two-piece design rendered it suitable for all sterilization techniques. PMID- 12566037 TI - Increased interleukin-6 in aqueous humor of a patient with lung-metastatic intraocular adenocarcinoma. AB - PURPOSE: We report increased interleukin (IL)-6 level in aqueous humor of a case of lung adenocarcinoma with metastasis to iris and anterior chamber. DESIGN: Interventional case report. METHODS: In one case and 11 healthy cataract patients as controls, we checked the levels of IL-6, IL-2, and tumor necrosis factor (TNF) alpha in aqueous humor and serum by enzyme-linked immunosorbent assay. RESULTS: Levels of these three cytokines in serum were almost the same as with the levels in aqueous humor in the control group. The IL-6 concentrations ranged from 12.6 to 27.0 pg/ml (mean, 23.5 pg/ml) in serum and 15.0 to 43.1 pg/ml (mean, 22.4 pg/ml) in aqueous humor. There was no statistically significant difference between them. Compared with the control, the IL-6 concentration in aqueous humor of this patient, 282.0 pg/ml, increased more than 10-fold. CONCLUSION: This is a report describing elevated IL-6 level in the aqueous humor of a patient with intraocular metastasis from lung adenocarcinoma. PMID- 12566038 TI - Choroidal metastasis from adenoid cystic carcinoma of the lung. AB - PURPOSE: We report a case of a choroidal metastasis from an adenoid cystic carcinoma of the lung. Interventional case report. DESIGN: A 40-year old man, 9 months' status postresection of a bronchial adenoid cystic carcinoma, was diagnosed by clinical evaluation and fine-needle aspiration biopsy and treated with palladium-103 ophthalmic plaque brachytherapy. RESULTS: This unusual patient with uveal metastasis from adenoid cystic carcinoma presented with decreased vision in the right eye and a diaphanous amelanotic choroidal tumor in the superotemporal macula. Ultrasound revealed a dome-shaped tumor that measured 4.0 mm in apical height and 11 x 10 mm in basal diameter. Fluorescein angiography revealed a double circulation and late intense subretinal fluorescence. Plaque brachytherapy provided local control and preservation of the eye. CONCLUSION: Bronchogenic adenoid cystic carcinoma can metastasize to the choroid. PMID- 12566039 TI - Continuous outflow devices for macular translocation with 360-degree retinotomy. AB - PURPOSE: Separation of the retina from retinal pigment epithelium is the most difficult step in macular translocation surgery. To complete this critical step, we have developed two specially designed continuous outflow needles, a rigid needle for creating retinal detachment and a liquid perfluorocarbon injection needle with a soft tip. DESIGN: Experimental study, new instrument development. METHODS: To permit continuous outflow during separation of the retina from the retinal pigment epithelium, the shaft of the continuous outflow needle is smaller than 20 gauge in diameter to allow continuous flow of fluid from the vitreous cavity. RESULTS: These instruments have been used successfully in 54 eyes that underwent macular translocation with 360-degree retinotomy. The rigid needle creates safe retinal detachment by keeping intraocular pressure lower, which is strictly needed for this step. The injection needle facilitates injection of perfluorocarbon liquid while manipulating the retina gently with its soft tip. CONCLUSION: Continuous outflow instruments are useful tools because they facilitate critical steps in the delicate process of macular translocation surgery. PMID- 12566040 TI - Submacular deposition of triamcinolone acetonide after triamcinolone-assisted vitrectomy. AB - PURPOSE: We describe a case demonstrating a submacular deposition of triamcinolone acetonide (TA) after a TA-assisted vitrectomy for retinal detachment. DESIGN: Interventional case report. METHODS: A 48-year-old Japanese man with rhegmatogenous retinal detachment in his left eye underwent a TA assisted vitrectomy, endolaser photocoagulation, and sulfur hexafluoride (SF(6)) gas tamponade. RESULTS: At the end of the surgery and the day after undergoing vitrectomy, the deposition of TA was observed between the retinal pigment epithelium and neurosensory retina in the submacular area. These TA granules disappeared after 2 weeks. Two months after the operation, the retina was observed to be successfully attached and no abnormality was observed in the macula. The patient's visual acuity improved to 20/16, and no ophthalmoscopic or functional damage was observed. CONCLUSION: No apparent adverse effect was found in this case demonstrating a submacular deposition of TA. PMID- 12566041 TI - Intravitreal triamcinolone for refractory pseudophakic macular edema. AB - PURPOSE: To evaluate the efficacy of intravitreal triamcinolone in refractory pseudophakic cystoid macular edema. DESIGN: A prospective, interventional case series. METHODS: Three eyes of three patients with longstanding pseudophakic cystoid macular edema following uncomplicated cataract surgery, refractory to any medication, were treated with 8 mg of intravitreal triamcinolone. All three eyes were evaluated before injection and throughout follow-up with the Early Treatment Diabetic Retinopathy Study's visual acuity chart, fluorescein angiography, and macular mapping using optical coherence tomography. RESULTS: A month after intravitreal triamcinolone injection, a dramatic decrease in macular thickness was noted by optical coherence tomography in all three eyes (from a mean of 502 233 microm). Mean improvement in visual acuity was 3.7 Snellen lines. Two to 4 months after triamcinolone injection, however, the edema recurred in all cases, to the same degree as before the injection, combined with a decrease in vision. Two eyes underwent a second injection of triamcinolone, and macular thickness decreased, but the edema again recurred 3 months after injection. CONCLUSION: Intravitreal injection of triamcinolone induces striking regression, within 1 month, of chronic refractory macular edema. This regression appears to be transient, however, even after a second injection. PMID- 12566042 TI - Ocular tilt reaction due to a mesencephalic lesion in juvenile polyarteritis nodosa. AB - PURPOSE: To describe a case of ocular tilt reaction caused by vasculitic lesions in the midbrain in a child with polyarteritis nodosa. DESIGN: Observational case report. METHODS: A 5-year-old girl with a chronic illness developed diplopia associated with a left head tilt, right hypertropia, torsional nystagmus, slowed vertical saccades and poor convergence. Fundoscopic examination demonstrated conjugate leftward torsion of the eyes consistent with a sustained ocular tilt reaction.Renal angiography confirmed polyarteritis nodosa and cerebral magnetic resonance imaging demonstrated mesencephalic pathology. CONCLUSIONS: Polyarteritis nodosa is a difficult condition to diagnose in a child and can cause brainstem lesions. This rare case of ocular tilt reaction of midbrain origin highlights that a sustained head tilt in a child can be due to brainstem pathology, rather than a fourth nerve palsy. PMID- 12566043 TI - Ipsilateral globe penetration and transient contralateral amaurosis following retrobulbar anesthesia. AB - PURPOSE: To report a rare complication of retrobulbar anesthesia with ipsilateral globe penetration and transient contralateral amaurosis. DESIGN: Interventional case report. METHODS: A 63-year-old woman complained of vision loss in the right eye immediately following cataract surgery on the left eye. RESULTS: Right eye vision decreased to no light perception with clear media and normal fundus. The vision recovered to baseline in 12 hours. Left eye vision was checked and demonstrated only light perception. Fundus examination disclosed preretinal and vitreous hemorrhage. During vitrectomy of the left eye, a penetrating wound below the optic disk with retinal detachment was found. CONCLUSION: The ipsilateral globe penetration wound depicts the mechanism of contralateral amaurosis following retrobulbar anesthesia of the case. PMID- 12566044 TI - Optic neuropathy produced by a compressed mucocele in an Onodi cell. AB - PURPOSE: To describe a patient with compressive optic neuropathy caused by a mucocele in an Onodi cell. DESIGN: Interventional case report. METHODS: A 73-year old man complained of sudden visual loss in the left eye 10 days after cataract surgery. Coronal computed tomography showed a homogeneous mass in an Onodi cell. Magnetic resonance imaging revealed an oval-shaped lesion that compressed the left optic nerve. RESULTS: After endoscopic microsurgery of the paranasal sinus, visual acuity in the left eye improved. Histopathological examination of the excised specimen showed a mucocele. CONCLUSIONS: Ophthalmologists should be aware that the mucocele in the Onodi cell, an anatomical variation of the paranasal sinus, may compress the optic nerve. PMID- 12566045 TI - Bone in the eye. AB - PURPOSE: To report a case of choroidal osteoma presenting with massive subretinal hemorrhage not associated with choroidal neovascularisation (CNV). DESIGN: Case report. METHODS: An 18-year-old man presented with sudden loss of vision in the right eye following competitive swimming. He was found to have a massive subretinal hemorrhage involving the macula. RESULTS: Following resolution of the hemorrhage, the patient was found to have an underlying choroidal osteoma. There was no evidence of choroidal neovascularisation clinically and angiographically. He regained his normal vision. CONCLUSIONS: Choroidal osteoma presenting with massive subretinal hemorrhage not associated with underlying choroidal neovascularization need not result in poor visual outcome. PMID- 12566046 TI - A novel mutation in the OPA1 gene in a Japanese patient with optic atrophy. AB - PURPOSE: To report a novel mutation of the OPA1 gene in a Japanese patient with optic atrophy and to describe the clinical features of the patient. DESIGN: Observational case report. METHODS: Genomic DNA was extracted from leukocytes of four unrelated Japanese patients with optic atrophy. All the exons and splice sites of the OPA1 gene were amplified by polymerase chain reaction and directly sequenced. RESULTS: One patient with optic atrophy had a heterozygous Arg445His mutation in the OPA1 gene. The Arg445His mutation was detected neither in 110 control subjects nor in the patient's healthy family members. CONCLUSIONS: A novel mutation of the OPA1 gene, similar to those reported in Western countries, was detected in a Japanese patient with optic atrophy. Mutations of the OPA1 gene may contribute to the development of optic nerve atrophy in Japanese cases of optic atrophy. PMID- 12566047 TI - Toxic effect of indocyanine green on retinal pigment epithelium related to osmotic effects of the solvent. PMID- 12566049 TI - Pupillary response to four concentrations of pilocarpine in normal subjects. PMID- 12566051 TI - Misspelling of Peters anomaly. PMID- 12566065 TI - Characterization and function of mitochondrial nitric-oxide synthase. AB - The mitochondrial production of nitric oxide is catalyzed by a nitric-oxide synthase. This enzyme has the same cofactor and substrate requirements as other constitutive nitric-oxide synthases. Its occurrence was demonstrated in various mitochondrial preparations (intact, purified mitochondria, permeabilized mitochondria, mitoplasts, submitochondrial particles) from different organs (liver, heart) and species (rat, pig). Endogenous nitric oxide reversibly inhibits oxygen consumption and ATP synthesis by competitive inhibition of cytochrome oxidase. The increased K(m) of cytochrome oxidase for oxygen and the steady-state reduction of the electron chain carriers provided experimental evidence for the direct interaction of this oxidase with endogenous nitric oxide. The increase in hydrogen peroxide production by nitric oxide-producing mitochondria not accompanied by the full reduction of the respiratory chain components indicated that cytochrome c oxidase utilizes nitric oxide as an alternative substrate. Finally, effectors or modulators of cytochrome oxidase (the irreversible step in oxidative phosphorylation) had been proposed during the last 40 years. Nitric oxide is the first molecule that fulfills this role (it is a competitive inhibitor, produced at a fair rate near the target site) extending the oxygen gradient to tissues. PMID- 12566066 TI - Quantification of 8-iso-prostaglandin-F(2alpha) and 2,3-dinor-8-iso-prostaglandin F(2alpha) in human urine using liquid chromatography-tandem mass spectrometry. AB - Quantification of 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2alpha)) has been suggested to be a reliable indicator of lipid peroxidation that may be related to in vivo free radical generation, oxidative damage, and antioxidant deficiency. We have developed a LC-MS/MS method to quantify 8-iso- PGF(2alpha) and its dinor metabolite, 2,3-dinor-8-iso-prostaglandin F(2alpha) (2,3-dinor-8-iso PGF(2alpha)), in human urine samples. After an initial purification step using an automated C18 solid phase extraction procedure, the urine sample was injected directly into a liquid chromatography (LC) system and detected with tandem mass spectrometry. The detection limit of the assay was 9 pg for 8-iso-PGF(2alpha) and 3 pg for 2,3-dinor-8-iso-PGF(2alpha) with both inter- and intraday variations of less than 12%. The inaccuracies were less than 3% for both analytes at three different levels. The urinary excretion rate of 2,3-dinor-8-iso-PGF(2alpha) was higher than that of 8-iso-PGF(2alpha), and changed in proportion to the parent compound (R = 0.70, n = 60). Values obtained with this method showed good linear correlation to duplicate 8-iso-PGF(2alpha) measurements performed with GCMS (R = 0.97, n = 15). The mean excretion rates of 8-iso-PGF(2alpha) and 2,3-dinor-8-iso PGF(2alpha) were significantly higher in smokers than in nonsmokers (0.53 +/- 0.37 vs. 0.25 +/- 0.15 microg/g creatinine, p = 0.002 for 8-iso-PGF(2alpha) and 8.9 +/- 3.8 vs. 4.6 +/- 2.6 microg/g creatinine, p = 0.003 for 2,3-dinor-8-iso PGF(2alpha), respectively). The excellent accuracy, reproducibility, and high throughput of this method should permit it to be used in large clinical studies and standard clinical laboratories. PMID- 12566067 TI - Icodextrin-induced lipid peroxidation disrupts the mesothelial cell cycle engine. AB - Fluids commonly used for peritoneal dialysis hold poor biocompatibility vis a vis the peritoneal membrane, basically due to the presence of osmotic agents. When rat mesothelium was exposed to glucose-enriched dialysis solutions for 2 h in vivo, an early and short-lived acceleration of cell life cycle was observed, which, after 30 d of exposure, resulted in a depopulated monolayer of senescent cells. These changes appear to result from persistent oxidative stress due to continuous exposure to high concentration of glucose and to substances generated by the Maillard reaction. Long-term exposure (30 d) of the peritoneal mesothelium to 7.5% icodextrin resulted in a depopulated monolayer consisting mostly of senescent cells, which, additionally, showed atypical nuclear changes and atypical mitosis suggesting DNA damage. These changes coincided with substantial lipid peroxidation, starting immediately after the introduction of the icodextrin solution into the rat's abdominal cavity. So far, the currently used osmotic agents in peritoneal dialysis fluids induce substantial oxidative injury to the exposed monolayer in vivo. Use of high concentrations of glucose results in premature senescence of the exposed cell population. The 7.5% icodextrin dialysis fluid induces through lipid peroxidation substantial genomic damage, which, in turn, sets the biological mechanisms leading to protective cellular suicide in motion. PMID- 12566068 TI - Superoxide-dependence of the short chain sugars-induced mutagenesis. AB - Short chain sugars such as glycolaldehyde are produced at the initial stages of nonenzymatic glycosylation. Because their carbonyl groups cannot be blocked by cyclization, such compounds tautomerize to enediols, which are prone to autoxidation. Superoxide radical serves as an initiator and a propagator of this autoxidation. The biological importance of the involvement of superoxide in sugar autoxidation in vivo was examined using superoxide dismutase (SOD)-deficient and SOD-replete strains of Escherichia coli. Glycolaldehyde, glyceraldehyde, and dihydroxyacetone greatly enhanced the mutation rates in SOD-deficient E. coli. The effect was oxygen-dependent and was suppressed by SOD or by a SOD mimetic. The mutagenic effect of glycolaldehyde coincided with intracellular accumulation of glyoxal, a product of glycolaldehyde autoxidation. PMID- 12566069 TI - Molecular ordering of ROS production, mitochondrial changes, and caspase activation during sodium salicylate-induced apoptosis. AB - Salicylates and nonsteroidal anti-inflammatory drugs (NSAIDs) induce apoptosis in a variety of cancer cells, including those of colon, prostate, breast, and leukemia. We examined the effects of sodium salicylate (NaSal) on reactive oxygen species (ROS) production and the association of these effects with apoptotic tumor cell death. We demonstrate that NaSal mediates ROS production followed by a decrease in mitochondrial membrane potential (deltapsi(m)), release of cytochrome c, and activation of caspase-9 and caspase-3. However, expression of Bcl-2 or Bcl x(L) prevents ROS production and subsequent loss of deltapsi(m), thereby inhibiting apoptotic cell death. The presence of ROS scavengers and an inhibitor of NADPH oxidase or expression of a dominant negative form of Rac1 blocks ROS production, deltapsi(m) collapse, and the subsequent activation of caspases. These observations indicate that NaSal mediates ROS production critical in the triggering of apoptotic tumor cell death through a Rac1-NADPH oxidase-dependent pathway. Our data collectively imply that NaSal-induced ROS are key mediators of deltapsi(m) collapse, which leads to the release of cytochrome c followed by caspase activation, culminating in tumor apoptosis. PMID- 12566070 TI - Metallothionein 2A induction by zinc protects HEPG2 cells against CYP2E1 dependent toxicity. AB - Zinc has been shown to have antioxidant actions, which may be due, in part, to induction of metallothionein (MT). Such induction can protect tissues against various forms of oxidative injury because MT can function as an antioxidant. The objective of this study was to investigate if zinc or MT induction by zinc could afford protection against CYP2E1-dependent toxicity. HepG2 cells overexpressing CYP2E1 (E47cells) were treated with 60 microM arachidonic acid (AA), which is known to be toxic to these cells by a mechanism dependent on CYP2E1, oxidative stress, and lipid peroxidation. E47 cells were preincubated overnight in the absence or presence of metals such as zinc or cadmium that can induce MT. The culture medium containing the metals was removed, AA was added, and cell viability determined after 24 h incubation. Preincubation overnight with 150 microM zinc sulfate or 5 microM cadmium chloride induced a 20- to 30-fold increase of MT2A mRNA; high levels of MT2A mRNA were maintained during the subsequent challenge period with AA, even after the zinc was removed. MT protein levels were increased about 4- to 5-fold during the overnight preincubation with zinc and a 20- to 30-fold increase was observed 24 h after zinc removal during the AA challenge. The treatment with zinc was associated with significant protection against the loss of cell viability caused by AA in E47 cells. The zinc pretreatment protected about 50% against the DNA fragmentation, cell necrosis, the enhanced lipid peroxidation and increased generation of reactive oxygen species, and the loss of mitochondrial membrane potential induced by AA treatment in E47 cells. CYP2E1 catalytic activity and components of the cell antioxidant defense system such as glutathione (GSH), glutathione-S-transferase (GST), glutathione peroxidase (GPX), catalase, Cu,Zn superoxide dismutase (SOD), and MnSOD were not altered under these conditions. Zinc preincubation also protected the E47 cells against BSO-dependent toxicity. When E47 cells were coincubated with zinc plus AA for 24 h (i.e., zinc was not removed, nor was there a preincubation period prior to challenge with AA), AA toxicity was increased. Thus, zinc had a direct pro-oxidant effect in this model and an indirect antioxidant effect, perhaps via induction of MT. MT may have potential clinical utility for the prevention or improvement of liver injury produced by agents known to be metabolized by CYP2E1 to reactive intermediates and to cause oxidative stress. PMID- 12566071 TI - Beta-carotene suppresses UVA-induced HO-1 gene expression in cultured FEK4. AB - The ultraviolet region of sunlight causes a significant oxidative stress to human skin cells and modulates expression of a series of genes in dermal fibroblasts and other cell types. The human heme oxygenase 1 (HO-1) gene is strongly activated within the first hours that follow UVA irradiation of normal human dermal fibroblasts (FEK4) and this response is being used as a marker of oxidative stress in cells. It has been shown that the induction of this gene occurs via singlet oxygen ((1)O(2)) produced upon interaction of UVA radiation with an as yet undefined cellular chromophore. Carotenoids, as the most potent singlet oxygen quenchers in nature, are expected to effectively suppress the UVA induced HO-1 gene activation in human cells. In this study, we measured the suppression of UVA-induced levels of HO-1 mRNA after the addition of a series of six all-trans-beta-carotene concentrations (0.07, 0.2, 0.8, 2.3, 8.0, and 21 microM) to the culture medium of exponentially growing FEK4 cells. The corresponding levels of beta-carotene uptake and apo-carotenal formation were measured following HPLC separation. The results of this study show a concentration-dependent suppression of UVA- (250 kJ/m(2)) induced transcriptional activation of HO-1 in exponentially growing FEK4 cells by beta-carotene. Suppression occurred at concentrations that have been observed in human plasma after dietary supplementation with beta-carotene. PMID- 12566072 TI - DPI induces mitochondrial superoxide-mediated apoptosis. AB - The iodonium compounds diphenyleneiodonium (DPI) and diphenyliodonium (IDP) are well-known phagocyte NAD(P)H oxidase inhibitors. However, it has been shown that at high concentrations they can inhibit the mitochondrial respiratory chain as well. Since inhibition of the mitochondrial respiratory chain has been shown to induce superoxide production and apoptosis, we investigated the effect of iodonium compounds on mitochondria-derived superoxide and apoptosis. Mitochondrial superoxide production was measured on both cultured cells and isolated rat-heart submitochondrial particles. Mitochondria function was examined by monitoring mitochondrial membrane potential. Apoptotic pathways were studied by measuring cytochrome c release and caspase 3 activation. Apoptosis was characterized by detecting DNA fragmentation on agarose gel and measuring propidium iodide- (PI-) stained subdiploid cells using flow cytometry. Our results showed that DPI could induce mitochondrial superoxide production. The same concentration of DPI induced apoptosis by decreasing mitochondrial membrane potential and releasing cytochrome c. Addition of antioxidants or overexpression of MnSOD significantly reduced DPI-induced mitochondrial damage, cytochrome c release, caspase activation, and apoptosis. These observations suggest that DPI can induce apoptosis via induction of mitochondrial superoxide. DPI-induced mitochondrial superoxide production may prove to be a useful model to study the signaling pathways of mitochondrial superoxide. PMID- 12566073 TI - External alternative NADH dehydrogenase of Saccharomyces cerevisiae: a potential source of superoxide. AB - Three rotenone-insensitive NADH dehydrogenases are present in the mitochondria of yeast Saccharomyces cerevisiae, which lack complex I. To elucidate the functions of these enzymes, superoxide production was determined in yeast mitochondria. The low levels of hydrogen peroxide (0.10 to 0.18 nmol/min/mg) produced in mitochondria incubated with succinate, malate, or NADH were stimulated 9-fold by antimycin A. Myxothiazol and stigmatellin blocked completely hydrogen peroxide formation with succinate or malate, indicating that the cytochrome bc(1) complex is the source of superoxide; however, these inhibitors only inhibited 46% hydrogen peroxide formation with NADH as substrate. Diphenyliodonium inhibited hydrogen peroxide formation (with NADH as substrate) by 64%. Superoxide formation, determined by EPR and acetylated cytochrome c reduction in mitochondria was stimulated by antimycin A, and partially inhibited by myxothiazol and stigmatellin. Proteinase K digestion of mitoplasts reduced 95% NADH dehydrogenase activity with a similar inhibition of superoxide production. Mild detergent treatment of the proteinase-treated mitoplasts resulted in an increase in NADH dehydrogenase activity due to the oxidation of exogenous NADH by the internal NADH dehydrogenase; however, little increase in superoxide production was observed. These results suggest that the external NADH dehydrogenase is a potential source of superoxide in S. cerevisiae mitochondria. PMID- 12566074 TI - Interaction of singlet molecular oxygen with double fluorescent and spin sensors. AB - Double fluorescent and spin sensors were recently used to detect transient oxidants via simultaneous fluorescence change and production of the nitroxide radical detected by electron paramagnetic resonance. One such oxidant, singlet molecular oxygen ((1)O(2)), was detected in thylakoid membrane using these probes. In the present study, we investigated the total (physical and chemical) quenching of (1)O(2) phosphorescence by sensors composed of the 2,5-dihydro 2,2,5,5-tetramethyl-1H-pyrrole moiety attached to xanthene or dansyl fluorophores. We found that the quenching rate constants were in the range (2-7) x 10(7) M(-1)s(-1) in acetonitrile and D(2)O. Quenching of (1)O(2) is usually an additive process in which different functional groups may contribute. We estimated that the (1)O(2) quenching by the amine fragments was ca. one to two orders of magnitude lower than that for the complete molecules. Our data suggest that the incorporation of a fluorescent chromophore results in additional strong quenching of (1)O(2), which may in turn decrease the nitroxide yield via the (1)O(2) chemical path, possibly having an effect on quantitative interpretations. We have also found that probes with the dansyl fluorophore photosensitized (1)O(2) upon UV excitation with the quantum yield of 0.087 in acetonitrile at 366 nm. This result shows that care must be taken when the dansyl-based sensors are used in experiments requiring UV irradiation. We hope that our results will contribute to a better characterization and wider use of these novel double sensors. PMID- 12566075 TI - The selenoprotein GPX4 is essential for mouse development and protects from radiation and oxidative damage insults. AB - Lipid peroxidation has been implicated in a variety of pathophysiological processes, including inflammation, atherogenesis, neurodegeneration, and the ageing process. Phospholipid hydroperoxide glutathione peroxidase (GPX4) is the only major antioxidant enzyme known to directly reduce phospholipid hydroperoxides within membranes and lipoproteins, acting in conjunction with alpha tocopherol (vitamin E) to inhibit lipid peroxidation. Here we describe the generation and characterization of GPX4-deficient mice by targeted disruption of the murine Gpx4 locus through homologous recombination in embryonic stem cells. Gpx4(-/-) embryos die in utero by midgestation (E7.5) and are associated with a lack of normal structural compartmentalization. Gpx4(+/-) mice display reduced levels of Gpx4 mRNA and protein in various tissues. Interestingly, cell lines derived from Gpx4(+/-) mice are markedly sensitive to inducers of oxidative stress, including gamma-irradiation, paraquat, tert-butylhydroperoxide, and hydrogen peroxide, as compared to cell lines derived from wild-type control littermates. Gpx4(+/-) mice also display reduced survival in response to gamma irradiation. Our observations establish GPX4 as an essential antioxidant enzyme in mice and suggest that it performs broad functions as a component of the mammalian antioxidant network. PMID- 12566076 TI - Use of the A(2A) adenosine receptor as a physiological immunosuppressor and to engineer inflammation in vivo. AB - Inflammation must be inhibited in order to treat, e.g., sepsis or autoimmune diseases or must be selectively enhanced to improve, for example, immunotherapies of tumors or the development of vaccines. Predictable enhancement of inflammation depends upon the knowledge of the "natural" pathways by which it is down regulated in vivo. Extracellular adenosine and A(2A) adenosine (purinergic) receptors were identified recently as anti-inflammatory signals and as sensors of excessive inflammatory tissue damage, respectively (Ohta A and Sitkovsky M, Nature 2001;414:916-20). These molecules may function as an important part of a physiological "metabolic switch" mechanism, whereby the inflammatory stimuli produced local tissue damage and hypoxia cause adenosine accumulation and signaling through cyclic AMP-elevating A(2A) adenosine receptors in a delayed negative feedback manner. Patterns of A(2A) receptor expression are activation- and differentiation-dependent, thereby allowing for the "acquisition" of an immunosuppressive "OFF button" and creation of a time-window for immunomodulation. Identification of A(2A) adenosine receptors as "natural" brakes of inflammation provided a useful framework for understanding how tissues regulate inflammation and how to enhance or decrease (engineer) inflammation by targeting this endogenous anti-inflammatory pathway. These findings point to the need of more detailed testing of anti-inflammatory agonists of A(2A) receptors and create a previously unrecognized strategy to enhance inflammation and targeted tissue damage by using antagonists of A(2A) receptors. It is important to further identify the contributions of different types of immune cells at different stages of the inflammatory processes in different tissues to enable the "tailored" treatments with drugs that modulate the signaling through A(2A) purinergic receptors. PMID- 12566077 TI - Apoptosis induction by the dual-action DNA- and protein-reactive antitumor drug irofulven is largely Bcl-2-independent. AB - The overexpression of Bcl-2 is implicated in the resistance of cancer cells to apoptosis. This study explored the potential of irofulven (hydroxymethylacylfulvene, HMAF, MGI 114, NSC 683863), a novel DNA- and protein reactive anticancer drug, to overcome the anti-apoptotic properties of Bcl-2 in HeLa cells with controlled Bcl-2 overexpression. Irofulven treatment resulted in rapid (12hr) dissipation of the mitochondrial membrane potential, phosphatidylserine externalization, and apoptotic DNA fragmentation, with progressive changes after 24hr. Bcl-2 overexpression caused marginal or partial inhibition of these effects after treatment times ranging from 12 to 48hr. Both Bcl-2-dependent and -independent responses to irofulven were abrogated by a broad spectrum caspase inhibitor. Despite the somewhat decreased apoptotic indices, cell growth inhibition by irofulven was unaffected by Bcl-2 status. In comparison, Bcl-2 overexpression drastically reduced apoptotic DNA fragmentation by etoposide, acting via topoisomerase II-mediated DNA damage, but had no effect on apoptotic DNA fragmentation by helenalin A, which reacts with proteins but not DNA. Irofulven retains its pro-apoptotic and growth inhibitory potential in cell lines that have naturally high Bcl-2 expression. Collectively, the results implicate multiple mechanisms of apoptosis induction by irofulven, which may differ in time course and Bcl-2 dependence. It is possible that the sustained ability of irofulven to induce profound apoptosis and to block cell growth despite Bcl-2 overexpression may be related to its dual reactivity with both DNA and proteins. PMID- 12566078 TI - Interactions of peroxynitrite and other nitrating substances with human platelets: the role of glutathione and peroxynitrite permeability. AB - Platelets labeled with 2',7'-dihydrodichlorofluorescein diacetate (DCF-DA) and stimulated with 50-400nM peroxynitrite (ONOO(-)) produced a rapid increase of the fluorescence signal at 523nm with good linearity and reproducibility. Platelet fluorescence was inhibited by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), suggesting that HCO(3)(-)/Cl(-) transporter mediated ONOO(-) transport into the platelets. Exposure of platelets to potassium superoxide, hydrogen peroxide, and sodium nitroprusside at concentrations of up to 100 microM did not generate a fluorescence signal. We also studied other nitrating compounds to establish the specificity of the DCF-DA-labeled platelet ONOO(-) assay. A rapid increase of fluorescence was observed when sodium hypochlorite (0.15 to 0.75mM) was added to platelets suspended in a buffered nitrite solution. Exposure of platelets to NO(2), nitroglycerin, and tetranitromethane produced a slow sustained increase of fluorescence. Endogenous glutathione appeared to be an essential factor in the generation of fluorescence by ONOO(-) and other nitrating compounds. We further studied other conditions that increased platelet fluorescence. Stimulation of platelets with thrombin (1U/mL) produced a rapid increase in fluorescence that corresponded to the formation of 20.5nmol ONOO(-) per 10(7) cells, whereas stimulation with collagen and arachidonic acid was without effect. Hypoxia of platelets for 20 and 40min followed by 5min of reoxygenation doubled the fluorescence from these platelets compared with control platelets. Thus, thrombin produced an effect that was likely due to the formation of ONOO(-) in platelets, whereas hypoxia-reoxygenation was likely to cause the formation of an active nitroglutathione-like molecule. PMID- 12566079 TI - Differential allosteric modulation by amiloride analogues of agonist and antagonist binding at A(1) and A(3) adenosine receptors. AB - The diuretic drug amiloride and its analogues were found previously to be allosteric modulators of antagonist binding to A(2A) adenosine receptors. In this study, the possibility of the allosteric modulation by amiloride analogues of antagonist binding at A(1) and A(3) receptors, as well as agonist binding at A(1), A(2A), and A(3) receptors, was explored. Amiloride analogues increased the dissociation rates of two antagonist radioligands, [3H]8-cyclopentyl-1,3 dipropylxanthine ([3H]DPCPX) and [3H]8-ethyl-4-methyl-2-phenyl-(8R)-4,5,7,8 tetrahydro-1H-imidazo[2,1-i]purin-5-one ([3H]PSB-11), from A(1) and A(3) receptors, respectively. Amiloride and 5-(N,N-dimethyl)amiloride (DMA) were more potent at A(1) receptors than at A(3) receptors, while 5-(N,N hexamethylene)amiloride (HMA) was more potent at A(3) receptors. Thus, amiloride analogues are allosteric inhibitors of antagonist binding at A(1), A(2A), and A(3) adenosine receptor subtypes. In contrast to their effects on antagonist occupied receptors, amiloride analogues did not affect the dissociation rates of the A(1) agonist [3H]N(6)-[(R)-phenylisopropyl]adenosine ([3H]R-PIA) from A(1) receptors or the A(2A) agonist [3H]2-[p-(2-carboxyethyl)phenyl-ethylamino]-5'-N ethylcarboxamidoadenosine ([3H]CGS21680) from A(2A) receptors. The dissociation rate of the A(3) agonist radioligand [125I]N(6)-(4-amino-3-iodobenzyl)adenosine 5'-N-methyluronamide ([125I]I-AB-MECA) from A(3) receptors was decreased significantly by amiloride analogues. The binding modes of amiloride analogues at agonist-occupied and antagonist-occupied receptors differed markedly, which was demonstrated in all three subtypes of adenosine receptors tested in this study. The effects of the amiloride analogues on the action of the A(3) receptor agonist were explored further using a cyclic AMP functional assay in intact CHO cells expressing the human A(3) receptor. Both binding and functional assays support the allosteric interactions of amiloride analogues with A(3) receptors. PMID- 12566080 TI - Impaired receptor-mediated endocytosis by the asialoglycoprotein receptor in ethanol-fed mice: implications for studying the role of this receptor in alcoholic apoptosis. AB - During receptor-mediated endocytosis (RME), extracellular molecules are internalized after being recognized and bound to specific cell surface receptors. In previous studies of the asialoglycoprotein receptor (ASGPR) in rats, we showed that ethanol impairs RME at multiple ASGPR sites. Ethanol administration has been shown to increase apoptosis, and we demonstrated increased sensitization to apoptotic induction in hepatocytes from ethanol-fed rats. Although a physiological role for the ASGPR has not been identified, investigators have shown its involvement in the uptake/clearance of apoptotic cells in vitro. This suggests a potential role for the ASGPR in the removal of apoptotic cells, and the recent availability of an ASGPR-deficient mouse strain provides an excellent opportunity to examine the role of the ASGPR during ethanol impairment. In this study, we examined ethanol-impaired RME in mice and began the characterization of ASGPR-deficient mice for use in ethanol studies. Similar to our findings with rats, ligand binding, internalization, and degradation were decreased 45-50% in hepatocytes from ethanol-fed wild-type mice. In ASGPR-deficient mice, these parameters did not vary among the chow-fed, pair-fed control, or ethanol groups and were negligible compared with those of wild-type mice. TUNEL analysis of liver sections showed an ethanol-induced increase in apoptotic bodies in all mouse strains with a significant difference in the receptor-deficient mice. Further, the livers of ASGPR-deficient mice had three times more apoptotic bodies, in all feeding groups, compared with wild-type mice. These results support the use of the ASGPR-deficient mouse model for studying ethanol-induced liver injury, specifically ethanol-induced apoptosis. PMID- 12566081 TI - Uroporphyrin accumulation in hepatoma cells expressing human or mouse CYP1A2: relation to the role of CYP1A2 in human porphyria cutanea tarda. AB - In experimental animals, CYP1A2 is absolutely required for the development of uroporphyria induced by treatment with polyhalogenated aromatic compounds or other compounds. Although the role of this CYP in clinical uroporphyria, porphyria cutanea tarda (PCT), is not clear, Cyp1a2(-/-) mice are resistant to the development of uroporphyria. Here, we compared the abilities of human and mouse CYP1A2 expressed in mouse hepatoma Hepa-1 cells to: (i) catalyze CYP1A2 dependent methoxyresorufin demethylase (MROD), and (ii) support uroporphyrin (URO) accumulation. Both CYP1A2 orthologs were expressed at similar levels as indicated by immunodetectable CYP1A2 proteins and MROD activities. URO accumulation was increased in cultures expressing either ortholog when supplemented with 5-aminolevulinic acid, the porphyrin precursor. Cells expressing mouse CYP1A2 produced more URO than cells expressing human CYP1A2. The results indicate that human CYP1A2 can support URO accumulation in hepatoma cells and thus may play a role in human PCT. PMID- 12566082 TI - The voltage- and time-dependent blocking effect of trifluoperazine on T lymphocyte Kv1.3 channels. AB - Phenothiazines are well-known calmodulin inhibitors that interact with many receptors and channels including a variety of potassium channels. In this study, we report a blocking effect of trifluoperazine (TFP) on voltage-gated Kv1.3 channels expressed in human T lymphocytes. Application of TFP in the concentration range from 1 to 20 microM reduced the current amplitude to about a half of the control value. The currents were blocked to less than 0.05 of the control value at 50 microM TFP concentration. The blocking effect was accompanied by a substantial increase in the current inactivation rate, whereas the activation rate and the steady-state activation and inactivation were not changed significantly. The blocking effect of TFP was voltage dependent being most potent at +60mV and least potent at -20mV. The blocking effect of TFP on the currents and the recovery from block was time dependent. Other calmodulin antagonists: tamoxifen (TMX) and thioridazine also inhibited the channels at micromolar concentrations. The effects exerted by TMX and thioridazine resembled the inhibitory effect of TFP. The blocking effect of thioridazine was time dependent and appeared to be more potent that the inhibition by TFP and TMX. TFP, TMX and thioridazine inhibited the activity of Kv1.3 channels only when applied extracellularly. The inhibitory effect of all the compounds was reversible. The possible physiological significance of the current inhibition is discussed. PMID- 12566083 TI - Selective activation of deoxycytidine kinase by thymidine-5'-thiosulphate and release by deoxycytidine in human lymphocytes. AB - Deoxycytidine kinase (dCK) catalyses the rate-limiting step of the salvage of three natural deoxyribonucleosides as well as several therapeutic nucleoside analogues, which in turn can enhance its enzymatic activity [Biochem Pharmacol 56 (1998) 1175], improving the efficacy of the cytostatic therapy. Here, we measured the effect of the 5'-thiosulphate (5'-TS) derivatives of four deoxyribonucleosides (deoxyadenosine, deoxycytidine (dCyd), azidothymidine, thymidine) and two ribonucleosides (ribopurine, ribouridine (Urd)) on the activity of the two main salvage deoxynucleoside kinases, and on the salvage of dCyd and deoxythymidine (dThd). It turned out that only 2'-deoxythymidine-5' thiosulphate (dThd-5'-TS) can potentiate the dCK activity, without influencing the thymidine kinase isoenzymes during short-time treatments of human peripheral blood and tonsillar lymphocytes. The enhancement of dCK activity by dThd-5'-TS can be reversed by dCyd, but dThd had no effect on the enzyme activation in cells. Neither dThd-5'-TS nor Urd-5'-TS had any effect on the dCK and thymidine kinase activities tested in cell-free extracts. The stimulation of dCK activity in cells was accompanied by an imbalance in the dThd and dCyd metabolism. The incorporation of 3H-dThd into DNA was suppressed by 90% in cells by dThd-5'-TS, while Urd-5'-TS only slightly influenced the same process. The 3H-dCyd incorporation into DNA was inhibited only to 50% of the control, while the 3H dCyd labelling of the nucleotide fraction was enlarged in dThd-5'-TS-treated cells, as a consequence of the increased dCK activity. We suggest that the enhancement of dCK activity is a compensatory mechanism in cells that might be induced by different "inhibitors" of DNA synthesis leading to damage of DNA. The increased dCK activity is able to supply the repair of DNA with dNTPs in quiescent cells; this suggestion seems to be supported by the counteracting effect of extracellular dCyd, too. PMID- 12566084 TI - Activation of deoxycytidine kinase by UV-C-irradiation in chronic lymphocytic leukemia B-lymphocytes. AB - Deoxycytidine kinase (dCK), a key enzyme of the deoxynucleoside salvage pathway, might have a preponderant role in DNA synthesis in resting chronic lymphocytic leukemia B-lymphocytes. In these cells, two important enzymes in deoxynucleoside triphosphate production, ribonucleotide reductase and thymidine kinase (TK), both cell-cycle regulated, are indeed very weakly expressed. This study investigated the regulation of dCK activity in response to UV-C light, a condition which causes DNA lesions and DNA repair synthesis. We observed that activity of dCK in B-CLL cells was upregulated up to 3-fold, 30 min after irradiation with 30 J/m(2) UV-C, whereas TK activity was unchanged. Activation of dCK by UV-C light was caused neither by a change in concentration of a low molecular weight metabolite nor by an increase in the amount of dCK protein. Activation of dCK by UV-C was mimicked by H(2)O(2), markedly counteracted by N-acetylcysteine, a general antioxidant, and completely abolished by the growth factor receptor inhibitor suramin. Taken together, these results indicate that dCK activity is upregulated by UV-C light through a postranslational modification that may be initiated at the cell surface through oxidative mechanisms. Suramin also suppressed the increase in DNA repair synthesis elicited by UV-C irradiation, suggesting that upregulation of dCK activity could contribute to the normal completion of DNA repair synthesis elicited by UV light. PMID- 12566085 TI - Effect of arvanil (N-arachidonoyl-vanillyl-amine), a nonpungent anandamide capsaicin hybrid, on ion currents in NG108-15 neuronal cells. AB - The effects of arvanil (N-arachidonoyl-vanillyl-amine), a structural hybrid between capsaicin and anandamide, on ion currents in a mouse neuroblastoma and rat glioma hybrid cell line, NG108-15, were examined with the aid of the whole cell voltage-clamp technique. Arvanil (0.2-50 microM) caused an inhibition of voltage-dependent L-type Ca(2+) current (I(Ca,L)) in a concentration-dependent manner. Arvanil produced no change in the overall shape of the current-voltage relationship of I(Ca,L). The IC(50) value of arvanil-induced inhibition of I(Ca,L) was 2 microM. Arvanil (5 microM) could shift the steady-state inactivation curve of I(Ca,L) to a more negative potential by approximately 15mV. No effect of arvanil (20 microM) on delayed rectifier K(+) current (I(K(DR))) was observed; however, capsaicin (20 microM), glyceryl nonivamide (20 microM) and capsinolol (20 microM) suppressed it significantly. Arvanil (20 microM) caused a slight reduction in the amplitude of erg (ether-a-go-go-related) mediated K(+) current (I(K(erg))) without modifying the activation curve of this current, while capsaicin and glyceryl nonivamide were more effective in suppressing I(K(erg)). Under current-clamp configuration, arvanil decreased the firing frequency of action potentials. Arvanil-mediated inhibition of I(Ca,L) appeared to be independent of its binding to either vanilloid or cannabinoid receptors. The channel-blocking properties of arvanil may, at least in part, contribute to the underlying mechanisms by which it affects neuronal or neuroendocrine function. PMID- 12566086 TI - Diminished production of nitric oxide synthase cofactor tetrahydrobiopterin by rosiglitazone in adipocytes. AB - Increased nitric oxide (NO) synthesis has been proposed to participate in the generation of insulin resistance in adipose and muscle tissues. Therefore, we examined the potential rate-limiting role of tetrahydrobiopterin (BH4) in cytokine-induced NO synthesis, and the effect of peroxisome proliferator activated receptor-gamma (PPARgamma) activation using the insulin-sensitizer rosiglitazone on cytokine-induced BH4 synthesis in 3T3-L1 adipocytes. Our data indicate that modulated availability of the mandatory nitric oxide synthase (NOS) cofactor BH4 affected cytokine-induced NO generation. Semiquantitative linear range reverse transcription polymerase chain reaction (RT-PCR) analysis demonstrated that rosiglitazone not only reduced inducible nitric oxide synthase (iNOS) mRNA transcription, but also guanosine triphosphate cyclohydrolase (GTPCH), the rate-limiting and controlling step of BH4 synthesis. Accordingly, intracellular BH4 concentration was reduced by 45% following rosiglitazone treatment. Furthermore, we observed a transient inhibitory effect of natural PPARgamma ligand 15-deoxy-Delta(12,14)-prostaglandin J2 (15d-PJ2) on cytokine mediated iNOS and GTPCH induction. Thus, the inhibition of cytokine-induced NO synthesis by rosiglitazone is at least in part attributable to reduced availability of BH4, the synthesis of which might represent a potential new target in the treatment of type 2 diabetes and insulin resistance. PMID- 12566087 TI - Inhibition of ATP-sensitive K(+)-channels by a sulfonylurea analogue with a phosphate group. AB - Hypoglycaemic sulfonylureas initiate insulin secretion by direct inhibition of ATP-sensitive K(+)-channels in the pancreatic beta-cells. These channels are composed of two proteins, a pore-forming subunit (K(IR)6.2 in the case of beta cells) and a regulatory subunit, the sulfonylurea receptor (SUR). In the present study we characterised the interaction with SURs of the new sulfonylurea analogues 5-chloro-N-[2-(4-hydroxyphenyl)ethyl]-2-methoxybenzamide (compound I) and (4-[2-(5-chloro-2-methoxybenzamido)ethyl]phenyl)phosphate (compound II). Compounds I and II differ from the sulfonylurea analogue meglitinide only in so far as the carboxylic group of meglitinide is replaced by a hydroxyl group or a phosphate group, respectively. The binding affinities of compound II for the SUR subtypes SUR1 (identified in beta-cells) and SUR2A (identified in heart and skeletal muscle) were higher by 55 or 21-fold, respectively, than the corresponding affinities for compound I. In inside-out patch-clamp experiments compound II inhibited ATP-sensitive K(+)-channels of the SUR1/K(IR)6.2-type (characteristic of beta-cells) with an IC(50) value of 0.16 microM which is 6 fold lower than the corresponding value for meglitinide. These findings strongly support the conclusion that the interaction of sulfonylureas and acidic analogues with SURs is favoured by the anionic group of these drugs and that a phosphate group allows more efficient ligand interaction with SUR1 than a carboxylic group. PMID- 12566088 TI - Structure activity analysis of the pro-apoptotic, antitumor effect of nitrostyrene adducts and related compounds. AB - In the present study, we outlined the part of the molecule mediating the prominent pro-apoptotic effect of the Michael adduct of ascorbic acid with p chloro-nitrostyrene, a new synthetic phosphatase inhibitor. The nitrostyrene (NS) moiety was identified as the structure essential for apoptosis induction. NS and its ascorbic acid adducts displayed LC(50) values of 10-25 microM with no significant reduction of potency in okadaic acid resistant cells overexpressing the MDR1 P-glycoprotein. Induction of apoptosis by NS derivatives and the protein phosphatase 2A inhibitor cantharidic acid was proven by the analysis of caspase-3 activation and subsequent fragmentation of DNA. Further structure activity analysis revealed the necessity of the nitro group at the beta-position of the side chain. The pro-apoptotic potential of adducts of NS with pyrimidine- or pyridine-derivatives varied between NS and a progressive reduction in potency up to a nearly complete loss of cytotoxicity. Substitutions at the benzene core of NS suggested a prominent enhancement of toxicity only by substitutions at the 2- or 3-position. Heterocyclic aromatics can substitute for the benzene ring of NS albeit with a 2-3-fold reduced potency. In conclusion, nitrostyrene was identified as the core structure mediating the pro-apoptotic effect of a new synthetic phosphatase inhibitor. Further studies defined a nitrovinyl side chain attached to an aromatic ring as the pharmacophore structure of a new group of pro apoptotic agents. These observations present the basis for the development of a new group of anticancer drugs. PMID- 12566089 TI - Antioxidants significantly affect the formation of different classes of isoprostanes and neuroprostanes in rat cerebral synaptosomes. AB - Lipid peroxidation has been implicated in the pathogenesis of a number of diseases, including neurodegenerative disorders. Evidence that antioxidants can affect the clinical course of neurodegenerative diseases is limited. In the present study, we examined the ability of five common antioxidants or antioxidant combinations, alpha-tocopherol, gamma-tocopherol, ascorbic acid, GSH ethyl ester, and a combination of ascorbate and alpha-tocopherol, to modulate lipid peroxidation in peroxidizing rat cerebral synaptosomes, a well-characterized model of oxidant injury. In these studies, we quantified isoprostanes (IsoPs) derived from arachidonic acid as an index of whole tissue oxidation and neuroprostanes (NeuroPs) formed from docosahexaenoic acid as a marker of selective neuronal peroxidation. We report that these various antioxidants displayed markedly different capacities to inhibit IsoP and NeuroP formation with the most potent effects on IsoPs observed for ascorbate, GSH ethyl ester, and the alpha-tocopherol-ascorbate combination. alpha-Tocopherol was slightly less potent and gamma-tocopherol significantly less effective. The concentration-response relationships were significantly different for NeuroP formation with the antioxidants being significantly less potent than for IsoP generation. In particular, alpha-tocopherol did not inhibit NeuroP formation at concentrations up to 100 microM. We also determined that tocopherols, in particular alpha tocopherol, act in vitro as reducing agents to convert IsoP and NeuroP endoperoxides to reduced F-ring compounds, a finding we have observed previously in vivo in brain. These studies are of importance because they have further defined the role of antioxidants to modulate the formation of lipid peroxidation products in peroxidizing brain tissue. In addition, they suggest that alpha tocopherol may not be a particularly effective agent to inhibit oxidant stress in the terminal compartment of neurons in the central nervous system. PMID- 12566090 TI - Modulation of [3H] TBOB binding to the rodent GABAA receptor by simple disaccharides. AB - We have recently reported evidence that a simple beta-linked alkylated mannose reversibly increased the magnitude of GABA(A) receptor currents evoked in cultured rat pyramidal neurons whilst concomitantly reducing the incidence of spontaneous synaptic activity. In this present study, the effects of the simple beta-linked disaccharide, lactose was investigated using a [3H] TBOB (t-[3H] butylbicycloorthobenzoate) binding assay in adult rat forebrain and cerebellum membranes. Lactose elicited a significant potentiation of [3H] TBOB binding to well-washed forebrain and cerebellar membranes (mean E(max) values=367 and 287%; mean EC(50) values=1.5 and 30 microM, respectively, N=4). The alpha-linked disaccharides, maltose and sucrose also potentiated [3H] TBOB binding, but with 100-600-fold higher EC(50) values than lactose. The lactose-mediated potentiation of [3H] TBOB in the forebrain and cerebellum was completely abolished in the presence of 0.3 microM GABA. Over the concentration range in which significant potentiation of [3H] TBOB binding was detected, lactose elicited no significant effect upon [3H] flunitrazepam binding. This study demonstrated that lactose can modulate the GABA(A) receptor channel, allosterically coupled to the agonist site, but independent of the benzodiazepine site. Furthermore, lactose displayed differential effects upon forebrain and cerebellar GABA(A) receptors indicating that it may be a novel subtype selective agent. PMID- 12566091 TI - Effects of zinc ion on type A monoamine oxidase in monkey brain mitochondria. AB - The effects of ZnSO(4) on types A and B monoamine oxidase (MAO) isozymes in monkey brain mitochondria were investigated, in vitro. Type A MAO activity in monkey brain decreased to about 50% with 1 microM ZnSO(4) using serotonin as a substrate, and this inhibition was proportional to the concentration of ZnSO(4). ZnSO(4) had no effect, however, on type B MAO activity in monkey brain using beta phenylethylamine as a substrate. The inhibition by ZnSO(4) of type A MAO activity was competitive and reversible. ZnSO(4) did not inhibit either type A or type B MAO activity in rat brain mitochondria. Almost similar results were also obtained when ZnCl(2) was used, in vitro. These results indicate that the inhibiting action of zinc ion differs depending on animal species and organ. Type A MAO in monkey brain mitochondria was highly sensitive to zinc ion, while type B activity was less sensitive. PMID- 12566092 TI - Insulin and estrogen receptor ligand influence the FGF-2 activities in MCF-7 breast cancer cells. AB - From the MCF-7 cell line we have developed, a human mammary cancer cell subline with the same karyotype as the mother strain and named MCF-7(SF), able to grow in serum-free chemically defined medium. This cell subline was firstly used to analyze the effect of basic fibroblast growth factor (FGF-2) in estrogen-receptor positive human breast cancer cells. FGF-2 like estradiol is able to increase cell proliferation and pS2 expression but was also found to inhibit progesterone receptor (PR) expression. The anti-estrogen tamoxifen partly counteracts the effects of FGF-2 and to discriminate between its two main mediators (estrogen receptor vs. anti-estrogen binding site, AEBS) we compare the efficacies of pure anti-estrogen (ICI 182,780) and AEBS ligand (PBPE). It appears that pure anti estrogen counteracts cell growth and pS2 effects of FGF-2 since AEBS ligand inhibits the cell growth but has no activity on pS2 expression. Secondly, adding insulin (10(-6)M) in the culture medium induces a strong increase in cell proliferation, which then elicits an inhibitory effect of FGF-2 and addition of anti-estrogens, are less efficient to further decrease growth, since the effects of FGF-2 and anti-estrogens on pS2 expression are conserved. PMID- 12566093 TI - Ethanol consumption decreases the synthesis of the mannose 6-phosphate/insulin like growth factor II receptor but does not decrease its messenger RNA. AB - The mannose 6-phosphate/insulin-like growth factor II receptor (M6P/IGF-IIR) is a protein that facilitates the transport of acid hydrolases into the lysosome. We have shown that chronic ethanol consumption lowers the M6P/IGF-IIR content in rat hepatocytes. Here, we determined the steady-state level of mRNA encoding M6P/IGF IIR, as well as the rate of receptor synthesis, to ascertain whether the ethanol elicited reduction in receptor protein content is related to changes in either or both of these parameters. Rats were pair-fed the normal carbohydrate (NC) or low carbohydrate high-fat (LC) liquid diets containing either ethanol or isocaloric maltose-dextrin for 7-8 weeks. RNA was isolated from hepatocytes and from whole livers of these animals and subjected to reverse transcription-polymerase chain reaction (RT-PCR) to determine the mRNA levels encoding M6P/IGF-IIR. Hepatocytes isolated from these animals were also radiolabeled with Pro-mix L-[35S] in vitro cell labeling mix to measure incorporation into total cellular protein and the immunoprecipitated M6P/IGF-IIR protein. The steady-state levels of M6P/IGF-IIR mRNA in both hepatocytes and whole livers from ethanol-fed rats were the same as those from their respective controls regardless of whether they were fed the NC or the LC diets. Hepatocytes from ethanol-fed rats showed a 36% lower rate of total protein synthesis and an even greater reduction (70%) in receptor synthesis. When the relative rate of receptor synthesis was calculated, hepatocytes from ethanol-fed rats had a 53% lower relative rate of receptor synthesis compared with controls. Autoradiographic analysis of the immunoprecipitated receptor protein from ethanol-fed rats also indicated a 79% decline in the total M6P/IGF-IIR protein synthetic rate compared with pair-fed controls. We conclude that the ethanol-elicited reduction of M6P/IGF-IIR content was, in part, related to a concomitant reduction of receptor protein synthesis but not to a decline in its mRNA level. Thus, the ethanol-elicited decline in receptor protein synthesis may be due to defective M6P/IGF-IIR mRNA translation. PMID- 12566094 TI - Suppression of human monocyte interleukin-1beta production by ajulemic acid, a nonpsychoactive cannabinoid. AB - Oral administration of ajulemic acid (AjA), a cannabinoid acid devoid of psychoactivity, reduces joint tissue damage in rats with adjuvant arthritis. Because interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha) are central to the progression of inflammation and joint tissue injury in patients with rheumatoid arthritis, we investigated human monocyte IL-1beta and TNFalpha responses after the addition of AjA to cells in vitro. Peripheral blood and synovial fluid monocytes (PBM and SFM) were isolated from healthy subjects and patients with inflammatory arthritis, respectively, treated with AjA (0-30 microM) in vitro, and then stimulated with lipopolysaccharide. Cells were harvested for mRNA, and supernatants were collected for cytokine assay. Addition of AjA to PBM and SFM in vitro reduced both steady-state levels of IL-1beta mRNA and secretion of IL-1beta in a concentration-dependent manner. Suppression was maximal (50.4%) at 10 microM AjA (P<0.05 vs untreated controls, N=7). AjA did not influence TNFalpha gene expression in or secretion from PBM. Reduction of IL 1beta by AjA may help explain the therapeutic effects of AjA in the animal model of arthritis. Development of nonpsychoactive therapeutically useful synthetic analogs of Cannabis constituents, such as AjA, may help resolve the ongoing debate about the use of marijuana as medicine. PMID- 12566095 TI - Decoy receptor 3 (DcR3) is proteolytically processed to a metabolic fragment having differential activities against Fas ligand and LIGHT. AB - Fas ligand (FasL) and Fas receptor are members of the tumor necrosis factor (TNF) receptor and ligand family that play an important role in regulating apoptosis in normal physiology. Decoy receptor 3 (DcR3) is a novel member of the TNF receptor superfamily, which binds to and blocks the activities of the ligands FasL and LIGHT. We have demonstrated that DcR3 was degraded rapidly to a major circulating metabolic fragment after subcutaneous administration in primates and mice. This fragment was also generated in subcutaneous tissue homogenate in vitro. Mass spectrometry and N-terminal sequencing indicated that DcR3 was proteolytically cleaved between R218 and A219 in the primary sequence to yield the fragment DcR3(1-218). While retaining its ability to bind LIGHT and inhibit LIGHT-mediated activities, DcR3(1-218) no longer bound FasL and did not inhibit FasL-mediated apoptosis in vitro. The primary sequence of DcR3 was molecularly engineered, changing the arginine residue at position 218 to glutamine to generate an analog, DcR3(R218Q), which we termed FLINT (LY498919). We demonstrated that FLINT was more stable to proteolytic degradation in vitro and in vivo and maintained its activity against both soluble FasL and soluble LIGHT in vitro. As a result, the modification in the sequence of DcR3 to produce FLINT (LY498919) should result in a pharmacologically superior molecule in the therapeutic intervention of diseases in which the pathogenesis is linked to FasL-mediated apoptotic or inflammatory events. PMID- 12566096 TI - Delphinidin inhibits endothelial cell proliferation and cell cycle progression through a transient activation of ERK-1/-2. AB - Epidemiological studies have shown that a diet rich in fruits and vegetables might reduce the risk of cardiovascular diseases and protect against cancer by mechanisms that have not been elucidated yet. This study was aimed to define the effect of delphinidin, a vasoactive polyphenol belonging to the class of anthocyanin, on bovine aortic endothelial cells (BAECs) proliferation. Delphinidin inhibited serum- and vascular endothelium growth factor-induced BAECs proliferation. This antiproliferative effect of delphinidin, is triggered by ERK 1/-2 activation, independent of nitric oxide pathway and is correlated with suppression of cell progression by blocking the cell cycle in G(0)/G(1) phase. Furthermore, suppression of cell cycle progression is associated with the modulation of the mitogenic signaling transduction cascade. This includes over expression of caveolin-1 and p21(WAF1/Cip1) and down-expression of Ras and cyclin D1. In conclusion, the antiproliferative effect of delphinidin may be of importance in preventing both plaque development and stability in atherosclerosis and tumor dissemination in cancer. PMID- 12566097 TI - Osthole prevents anti-Fas antibody-induced hepatitis in mice by affecting the caspase-3-mediated apoptotic pathway. AB - Fas (Apo-1/CD95) ligand, which is a type II membrane protein, is a major inducer of apoptosis. Osthole is a coumarin derivative present in medicinal plants. The effect of osthole on hepatitis induced by anti-Fas antibody in mice was studied. Pretreatment of mice with osthole (10, 50, and 100 mg/kg, i.p.) prevented the elevation of plasma alanine aminotransferase (ALT) caused by anti-Fas antibody (175 microg/kg, i.v.). Administration of osthole to mice even at a dose of 10 mg/kg significantly inhibited of anti-Fas antibody-induced elevation of plasma ALT. Capase-3 is a cysteine protease, and treatment of mice with anti-Fas antibody caused an elevation of caspase-3 activity at 3.5 and 6 hr. Pretreatment of mice with osthole (100 mg/kg, i.p.) inhibited the elevation of caspase-3 activity caused by anti-Fas antibody. However, the addition of osthole (up to 10( 4)M) to a liver cytosol fraction isolated from mice treated with anti-Fas antibody did not inhibit caspase-3 activity in vitro. Thus, treatment of mice with osthole inhibited caspase-3 activity by an effect upstream of caspase-3 activation. The livers of mice treated with anti-Fas antibody contained apoptotic and dead cells; osthole attenuated the development of this apoptosis and cell death. The present results show that osthole prevented anti-Fas antibody-induced hepatitis by inhibiting the Fas-mediated apoptotic pathway. PMID- 12566098 TI - Indomethacin-induced free radical-mediated changes in the intestinal brush border membranes. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) cause small intestinal damage but the pathogenesis of this toxicity is not well established. Our earlier work has shown that villus enterocytes are most susceptible to the effects of indomethacin, a commonly used NSAID. This study looked at the acute effect of indomethacin on brush border membranes (BBM), which are present mainly in the villus cells and are in immediate contact with the contents of the small intestinal lumen. Evidence of oxidative stress was found in the mucosa of the small intestine of rats dosed with indomethacin, as indicated by increased activity of xanthine oxidase with corresponding decrease in the levels of several free radical scavenging enzymes. These changes were associated with an increase in peroxidation parameters in the BBM and a fall in the level of alpha tocopherol. These BBM also exhibited impairment in glucose transport. Significant changes were seen in the lipid composition of these membranes, with upregulation of an 85kDa isoform of phospholipase A(2). Pretreatment of animals with allopurinol, arginine or zinc protected against these effects of indomethacin. Thus this study suggests that in an acute model of indomethacin dosing there is impairment in structure and function of the BBM in enterocytes, with the effects possibly mediated by free radicals and phospholipases. PMID- 12566100 TI - Calcineurin, myocardial hypertrophy, and electrical remodeling. PMID- 12566101 TI - Human heart failure: our current STATus of knowledge. PMID- 12566102 TI - Outcome of clinical versus genetic family screening in hypertrophic cardiomyopathy with focus on cardiac beta-myosin gene mutations: prediction of clinical status--is molecular genetics a new tool for the management of hypertrophic cardiomyopathy in clinical practice? PMID- 12566103 TI - Myocardial blood flow in patients with hibernating myocardium. AB - The debate on whether resting myocardial blood flow (MBF) to hibernating myocardium is reduced or not has attracted a lot of interest and has contributed to stimulate new research on heart failure in patients with coronary artery disease (CAD). Positron emission tomography with oxygen-15 labeled water (H(2)(15)O) or nitrogen-13 labeled ammonia (13NH(3)) has been used for the absolute quantification of regional MBF in human hibernating myocardium. When hibernating myocardium is properly identified, i.e. a dysfunctional segment subtended by a stenotic coronary artery that improves function upon reperfusion, the following conclusions can be reached based on the available literature: (a) in the majority of these studies resting MBF in hibernating myocardium is not different from either flow in remote tissue in the same patient or MBF in normal healthy volunteers; (b) a reduction in MBF of approximately 20% compared to MBF in remote myocardium or age matched normal subjects has been demonstrated in a minority of truly hibernating segments; (c) hibernating myocardium is characterized by a severely impaired coronary flow reserve which improves after revascularization in parallel with contractile function. Thus, the pathophysiology of hibernation in humans is more complex than initially postulated. The recent evidence that repetitive ischemia in patients with coronary artery disease can be cumulative and lead to more severe and prolonged stunning, lends further support to the hypothesis that, at least initially, stunning and hibernation are two facets of the same coin. PMID- 12566104 TI - Insulin and myocardial blood flow. AB - The renaissance of glucose-insulin-potassium infusion (GIK) as a treatment of acute myocardial infarction both in diabetic and nondiabetic subjects has raised new interests to clarify the effects and mechanisms of insulin on myocardium. Although the action of insulin on substrate metabolism is quite well studied in heart, the cardiovascular effects were until recent years poorly known. Insulin induces skeletal muscle vasodilation mainly via the endothelium-dependent mechanism and appears to have an important role in normal vascular function. There is increasing amount of evidence that insulin acts as a vasodilatory hormone also in coronary arteries. Insulin enhances myocardial blood flow and decreases coronary vascular resistance in a dose-dependent manner in healthy subjects. Moreover, insulin is able to increase myocardial blood flow also in subjects who are characterized by coronary dysfunction such as subjects with obesity, type 1 diabetes and coronary artery disease. However, vasodilatory effect of insulin may be blunted in these patients. Since already very small increase in myocardial blood flow can reduce significantly myocardial ischemia, these vasodilatory actions of insulin in coronary arteries might partly contribute to beneficial effects of GIK therapy. On the other hand, in contrast to these acute beneficial effect of insulin, epidemiological studies have indentified chronic hyperinsulinemia, a common feature in subjects with insulin resistance to glucose uptake, as an independent risk factor for coronary artery disease. The present article review the physiological and pathophysiological role of insulin in cardiac vasculature and its clinical importance during myocardial ischemia and development of coronary artery disease. PMID- 12566105 TI - Overexpression of calcineurin in mouse causes sudden cardiac death associated with decreased density of K+ channels. AB - BACKGROUND: Overexpression of calcineurin in transgenic (TG) mice results in cardiac hypertrophy and unexpected deaths. METHODS AND RESULTS: None of the TG survived beyond 24 weeks (n=38) whereas all of the wildtype (WT, n=47) survived. Prolongation of repolarization preceded the development of sustained pleomorphic ventricular tachycardia and high degree atrioventricular block, which occurred during spontaneous sudden deaths. Since depolarization-activated K(+) channels contribute dominantly to repolarization in mice, we hypothesized that the TG would decrease these K(+) currents and that the in vivo administration of cyclosporin A (CsA), a calcineurin inhibitor, would reduce this effect. CsA reversed cardiac hypertrophy: capacitance measurements of WT left ventricular myocytes (127+/-7 pF; n=45) and CsA-treated TG (129+/-14 pF; n=17) were significantly lower than in placebo-treated TG (220+/-11 pF; n=41; P<0.001 by ANOVA). Independent of whether the data fit a bi- or a tri-exponential model, the density of I(tof) was significantly reduced in TG versus WT and CsA reversed this effect. While I(tos) and I(Kslow) were also reduced in TG, CsA does not reverse this change because long-term in vivo CsA treatment of WT also reduces I(tos) and I(Kslow.) To assess whether the decreased 'repolarization reserve' contributed to arrhythmogenesis, the residual I(Kr) was blocked by dofetilide precipitating pleomorphic ventricular tachycardias. CONCLUSION: Since the downregulation of I(tof) was observed with overexpression of calcineurin and was also reversed by the calcineurin inhibitor CsA, we conclude that downregulation of I(tof) is a consequence of calcineurin overexpression. PMID- 12566106 TI - Activation of signal transducer and activator of transcription (STAT) pathways in failing human hearts. AB - OBJECTIVES: The signal transduction pathways mediating the progression to failure have been intensively studied in a variety of in vitro and in vivo animal models. Recently, acute activation of the Janus kinases (JAKs) and signal transducers and activators of transcription (STATs) has been observed in the heart, but whether this is sustained in ischemic heart disease (IHD) or dilated cardiomyopathy (DCM) has not been previously addressed. METHODS: We assessed the tyrosine phosphorylation of STAT1, 3, 5 and 6 in ventricular samples of explanted human hearts with IHD (n=11) and DCM (n=9) as an indication of STAT activation. Samples from normal donor hearts (n=9) acted as controls. In parallel, we also assessed protein expression and phosphorylation of three major families of mitogen activated protein kinases (MAPKs); ERK, p38 MAPK and c-Jun NH(2)-terminal kinase (JNK). RESULTS: All STAT isoforms were significantly phosphorylated in DCM. In contrast, only the phosphorylation of STATs 1 and 5 were significantly enhanced in IHD. Expression of total STAT protein remained unchanged. For the MAPKs, significant phosphorylation of p38(MAPK) was only observed in IHD. In contrast, there was no change in ERK or JNK activation despite abundant protein expression. CONCLUSIONS: We have shown that different members of the STAT transcription factor family are chronically phosphorylated in the failing heart as a result of IHD (STAT1 and 5) or DCM (STAT1, 3, 5 and 6). In contrast, IHD but not DCM showed significant p38(MAPK) phosphorylation. Whilst the differences noted between IHD and DCM may reflect different initiating events, the common activation of STATs 1 and 5 suggests that these transcription factors may play a common role regulating the progression of heart failure. PMID- 12566107 TI - Outcome of clinical versus genetic family screening in hypertrophic cardiomyopathy with focus on cardiac beta-myosin gene mutations. AB - OBJECTIVE: Familial hypertrophic cardiomyopathy (FHC) is caused by mutations in genes encoding cardiac sarcomere proteins. Although available, genetic analyses are generally not used clinically. In the present study, we evaluated the outcome of clinical vs. genetic screening of family members with specific focus on mutations in the cardiac beta-myosin heavy chain (MYH7) gene. METHODS: A consecutive cohort of 68 FHC probands and their families (395 persons) of Danish origin was evaluated including patient- and family histories, physical examinations, electrocardiogram and echocardiography. Mutation screening was performed by a combination of single strand conformation/heteroduplex analysis and direct sequencing. RESULTS: Eight different MYH7 gene mutations were identified in nine (13%) families (96 persons). In eight (89%) of the families, major cardiac events had occurred. Myectomy or percutaneous septal alcohol ablation had been performed in a higher number of MYH7 probands i.e. in five of nine (56%) as compared to 10 of 59 (17%) (P<0.05) non-MYH7 mutation probands. Neither echocardiographic nor ECG findings were useful to distinguish MYH7 from non-MYH7 probands. Between adult MYH7 mutation-carriers (n=38) and their non carrier relatives (n=39), low sensitivity and specificity of the clinical diagnostic criteria tested were observed and minor clinical diagnostic criteria alone were not useful for identification of mutation carriers. By genetic screening of relatives with no or only minor hypertrophy on echocardiography, i.e. a priori possible mutation-carriers normally recommended clinical follow-up the diagnosis was excluded in 52 (83%) persons. In addition, six relatives with secondary hypertrophy were identified as non-carriers. CONCLUSION: Neither echocardiographic nor ECG findings were useful to distinguish MYH7 from non-MYH7 probands. Extension of screening to include genetic analyses offered a marked diagnostic advantage as compared to clinical screening alone in FHC families. PMID- 12566108 TI - Caveolin-1 and -3 dissociations from caveolae to cytosol in the heart during aging and after myocardial infarction in rat. AB - OBJECTIVE: Caveolins, the structural proteins of caveolae, modulate numerous signaling pathways including Nitric Oxide (NO) production. Among the caveolin family, caveolin-1 and -3 are mainly expressed in endothelial and muscle cells, respectively. In this study, we investigate whether (i) changes in caveolin abundance and/or distribution occur during cardiac aging and failure in rat, and (ii) the process could influence NO synthase (NOS) activity. METHODS: Using immunohistolabelling and Western blot approaches, expression and distribution of caveolins were analysed in adult (Ad), senescent (S-Sh) and myocardial infarction induced failing (S-MI) hearts. NOS3/caveolin-1 interactions were evaluated by immunoprecipitation assays. RESULTS: At the microscope level, caveolin-1 distribution in the endothelial cells was unchanged between the groups. Conversely the typical distribution of caveolin-3 in myocyte sarcolemma was dramatically altered in S-MI rats, resulting in a heterogeneous pattern throughout the septum. Total abundance of caveolin-1 and -3 remained stable whatever the group. In the fractions free of caveolae (Triton X-100 soluble), the levels of caveolin-1 alpha and -3 increased with aging (+20%, and +104%, P<0.05 versus Ad, respectively) and were further enhanced in S-MI (+25%, +30%, P<0.05, P<0.001 versus S-Sh respectively). In these fractions, NOS3/caveolin-1 alpha complexes increased as well. In addition, NOS activity was negatively correlated to caveolin-1 level in the cytosolic fractions. CONCLUSIONS: We demonstrate that dissociation of caveolin from caveolae is associated with aging and heart failure, the process being related to the decreased NOS activity. PMID- 12566109 TI - Effect of testosterone on post-myocardial infarction remodeling and function. AB - BACKGROUND: Men and women are differently affected by coronary artery disease, suggesting an important role of sex steroids. Moreover, testosterone (T) treatment is increasingly used in elderly males. Therefore, we examined effects of chronic anabolic T administration on left ventricular (LV) remodeling after myocardial infarction (MI). METHODS: Adult male rats were treated with intramuscular placebo, testosterone undecanoate (T), or were orchiectomized. After 2 weeks, animals underwent sham-operation (sham) or left coronary artery ligation. Left ventricular remodeling and function was assessed by serial magnetic resonance imaging (MRI) at weeks 2 and 8 and hemodynamic investigation at week 8. RESULTS: In sham operated animals T administration increased serum T levels and led to cardiac hypertrophy, but not to an upregulation of ANP mRNA. The alpha/beta-MHC ratio was significantly higher after T treatment due to an increase in alpha-MHC. As a potential mechanism for this "physiologic" form of hypertrophy, IGF-1 mRNA expression was significantly increased in T treated animals. After coronary artery ligation, infarct size and mortality were similar among the groups. Left ventricular hypertrophy was enhanced by T treatment. However, in vivo LV end-diastolic pressure and wall stress were decreased by T, whereas other hemodynamic parameters (mean arterial pressure, cardiac output, etc.) remained unchanged. CONCLUSION: Chronic anabolic T treatment led to a specific "physiologic" pattern of myocardial hypertrophy with a significant increase in LV weight, but without differences in ANP and with an upregulation in alpha/beta-MHC, possibly mediated by IGF-1. Testosterone treatment had no detrimental effects following MI. Reduced wall stress and LVEDP may even improve long-term outcome. PMID- 12566110 TI - Significance of matrix metalloproteinases in norepinephrine-induced remodelling of rat hearts. AB - OBJECTIVE: Norepinephrine (NE) induced hypertrophy and remodelling of the extracellular matrix (ECM) in the left ventricle (LV) of the rat heart with resulting fibrosis. However, there was no increased collagen deposition in the right ventricle (RV). To test the hypothesis that lack of RV fibrosis is the result of elevated cleavage of collagens we inhibited the activity of matrix metalloproteinases (MMP) by doxycycline (Doxy) and then measured function and collagen metabolism in the RV as compared to the LV. METHODS: Female Sprague Dawley rats were treated with 30 mg/kg per day doxycycline alone or in combination with i.v. infusion of NE (0.1 mg/kg per h). The activity of MMP-2 was increased both in the LV and RV after 3 days of NE infusion and reduced after concomitant doxycycline treatment which also caused inhibition when given alone. RESULTS: After 14 days of NE infusion in combination with doxycycline there was an additional increase in the NE-induced elevation of collagen accumulation in the LV (interstitial collagen fraction: NE-Doxy 1.797%, P<0.05 versus control and NE; NE 1.113%, P<0.05 versus control) and significant fibrosis in the RV (2.105%, P<0.05 versus control). This correlated with the prevention of the NE-induced elevation of RV systolic pressure (NE: 71.3 mmHg, P<0.05; NE-Doxy: 36.4 mmHg) and RV dP/dt(max) (NE: 5500 mmHg/s, P<0.05; NE-Doxy: 2550 mmHg/s). Also in the NE stimulated LV, the doxycycline-induced collagen accumulation was associated with reduced LV dP/dt(max) (NE-Doxy: 13169 mmHg/s; NE: 18849 mmHg/s, P<0.05). CONCLUSION: MMP inhibition leads to myocardial stiffness with negative functional consequences for the RV and LV in NE-treated rat hearts. PMID- 12566111 TI - Estrogen modulation of left ventricular remodeling in the aged heart. AB - OBJECTIVE: To investigate the effects of estrogen on left ventricle (LV) mass and collagen deposition, and on the expression of receptors for estrogen (ER alpha, ER beta) and Ang II (AT(2)R, AT(1)R) in the heart of aged female rats. METHODS: Aged ( approximately 12 months old) intact (n=7), ovariectomized plus placebo (OVX, n=7), and estrogen-replaced (E2, n=6) as well as young ( approximately 3 months old, n=4) female Sprague-Dawley rats were used in this study. After 1 month of treatment, the left ventricular weight/body weight ratio (LVW/BW), changes in myosin heavy chain expression (MHC), matrix metalloproteinase (MMP)-2 activity, the collagen I/III ratio, and the expression of ERs and Ang II receptors in the LV were evaluated. RESULTS: In aged rats, OVX increased LVW/BW associated with a higher expression of beta-MHC isoform, increased collagen I/III ratio, and decreased MMP-2 activity compared to intact rats. Furthermore, the OVX group had a decrease in ERs alpha and beta as well as AT(2)R but an increase in AT(1)R expression. Estrogen replacement prevented the effects of ovariectomy on heart remodeling as well as increased further expression of ER beta and decreased AT(1)R expression. CONCLUSION: Removal of ovarian hormones increased LV remodeling in the aged rat, which could be attenuated by estrogen replacement. Moreover, regulation of Ang II receptor expression could be a mechanism by which estrogen may modulate heart remodeling. PMID- 12566112 TI - Increased myocardial collagen and ventricular diastolic dysfunction in relaxin deficient mice: a gender-specific phenotype. AB - OBJECTIVE: To investigate cardiac phenotypes in mice deficient in the peptide hormone relaxin by gene targeting. METHODS: Echocardiography and cardiac catheterization were performed on male and female relaxin deficient (Rlx(-/-)) mice as well as heterozygous (Rlx(+/-)) and wildtype (Rlx(+/+)) littermates aged between 8 and 24 months. Collagen expression and content in the heart were analysed by real-time PCR, hydroxyproline assay and histology. RESULTS: Heart rate, blood pressures, left ventricular (LV) dimensions, fractional shortening and maximal and minimal dP/dt did not differ significantly between the three genotypes of either gender at any age. However, 8-10-month-old Rlx(-/-) males exhibited a greater transmitral flow velocity (A-wave) at the late LV diastolic phase. Male Rlx(-/-) mice aged between 12 and 24 months had significantly higher LV end-diastolic pressures, a 30% increase in atrial weight and 10-30% increases in lung and liver weights. Male mice also showed an age-dependent increase (P<0.01) in LV collagen content that was more pronounced in Rlx(-/-) than control littermates (P<0.01). Procollagen type-1 expression was also significantly higher in the LV of Rlx(-/-) males compared with either Rlx(+/-) or Rlx(+/+) males at 6, 9 and 12 months of age. Age-matched female Rlx(-/-) mice did not display any of these cardiac phenotypes seen in Rlx(-/-) males. CONCLUSIONS: Male Rlx(-/-) mice had impeded LV diastolic filling and increased atrial weights, most likely due to an increase in ventricular collagen content and chamber stiffness. These phenotypes in the Rlx(-/-) males were not observed in Rlx(-/-) females, indicating the importance of other gender-related factors in cardiovascular function. PMID- 12566113 TI - The cardioprotective and mitochondrial depolarising properties of exogenous nitric oxide in mouse heart. AB - OBJECTIVE: Nitric oxide (NO) is reported to be both protective and detrimental in models of myocardial ischaemia/reperfusion injury, which may be concentration dependent. Our objective was to characterise this dichotomy using the nitric oxide donor, S-nitroso N-acetyl penicillamine (SNAP) in isolated perfused mouse heart and isolated mouse cardiac mitochondria. METHODS: To determine the effect of nitric oxide concentration on myocardial viability, isolated mouse hearts were subjected to 35 min global ischaemia and 30 min reperfusion in the presence of SNAP (0.02-20 microM). To determine whether NO mediated protection was via opening of the putative mitochondrial K(ATP) channel and/or free radical synthesis, SNAP perfused hearts were also treated with the mitochondrial K(ATP) channel blocker, 5-hydroxy decanoate (5-HD) and the free-radical scavenger, N-(2 mercaptopropionyl)-glycine (MPG). This data was correlated with mitochondrial membrane potential (Delta Psi(m)), measured with the potentiometric dye, tetra methyl rhodium methyl ester (TMRM), in isolated mitochondria,by flow cytometry. RESULTS: SNAP dose-dependently attenuated infarct size, with maximal protection observed at 2 microM (17+/-4% versus controls 32+/-3%, P<0.01). At greater concentrations however, protection was lost with infarct sizes tending towards control at 20 microM (29+/-3%). These results were paralleled by changes in Delta Psi(m) in the isolated mitochondria: Delta Psi(m) depolarisation peaking with 1 microM SNAP (26+/-4% shift in TMRM fluorescence, P<0.01); at greater concentrations, this relationship was lost. The mitochondrial K(ATP) channel blocker, 5-HD, resulted in both abrogation of SNAP infarct size reduction and concomitant loss of Delta Psi(m) depolarisation in the mitochondria. MPG however did not influence the cardioprotective properties of SNAP. CONCLUSION: We demonstrate that nitric oxide can mediate cardioprotection in a dose-dependent fashion by an effect that may be related to Delta Psi(m). Both cardioprotection and Delta Psi(m) changes are sensitive to 5-HD and the cardioprotection appears independent of free-radical synthesis. PMID- 12566114 TI - Effects of N-(2-mercaptopropionyl)-glycine on mitochondrial function in ischemic reperfused heart. AB - OBJECTIVE: A possible mechanism for N-(2-mercaptopropionyl)-glycine (MPG) underlying the improvement of contractile function and mitochondrial activity of ischemic-reperfused rat hearts was examined. METHODS: Isolated, perfused hearts were subjected to 35 min ischemia-60 min reperfusion. At the end of ischemia or reperfusion, myocardial Na(+) content and mitochondrial oxygen consumption rate (OCR) were examined. The perfused heart was treated with 0.1-1 mM MPG for 30 min prior to ischemia or for the first 30 min of reperfusion. RESULTS: Ischemia increased myocardial Na(+) content (sodium overload) and decreased mitochondrial OCR. The left ventricular developed pressure (LVDP) of the untreated heart recovered to 19.8+/-3.8% of the preischemic value and the infarct area amounted to 23.3+/-1.7% of the left ventricle. The thiobarbiturate-reacting substance (TRS) was also increased in the reperfused, but not ischemic, myocardium. Pretreatment of the perfused heart with 0.3-1 mM MPG attenuated the ischemia induced sodium overload and decrease in the OCR. Pretreatment with the agent also enhanced the postischemic recovery of LVDP, attenuated reperfusion-induced increase in TRS, and reduced the infarct area. Although the postischemic treatment with MPG suppressed the increase in TRS in the reperfused myocardium, a LVDP recovery of reperfused hearts was not observed. Cardiac mitochondria were isolated and examined for the direct effect of MPG on their function. Incubation with either 12.5 mM sodium lactate or 1 microM phenylarsine oxide neither altered the mitochondrial membrane potential nor induced mitochondrial swelling, whereas incubation with a combination of these agents elicited the membrane potential depolarization and swelling. Incubation of mitochondria with 1 mM MPG attenuated these events. CONCLUSION: These results suggest that both attenuation of sodium overload and preservation of the mitochondrial function may largely contribute to cardioprotection of MPG in the ischemic-reperfused heart. PMID- 12566115 TI - Matrix metalloproteinase-2 mediates cytokine-induced myocardial contractile dysfunction. AB - OBJECTIVE: Pro-inflammatory cytokines depress myocardial contractile function by enhancing peroxynitrite production, yet the mechanism by which peroxynitrite does this is unknown. As matrix metalloproteinases (MMPs) can be activated by peroxynitrite and can proteolytically cleave troponin I in hearts, we determined whether this occurs in cytokine-induced myocardial dysfunction. METHODS: Isolated working rat hearts were perfused with buffer containing interleukin-1 beta, interferon-gamma, and tumor necrosis factor-alpha. RESULTS: Cytokines induced a marked decline in mechanical function during 60-120 min of perfusion. This decline was accompanied by increased myocardial inducible NO synthase activity and perfusate dityrosine (a marker of peroxynitrite), compared to control hearts. Before the decline in mechanical function there was enhanced MMP-2 activity in the perfusate. This was accompanied by decreased tissue levels of MMP-2, tissue inhibitor of matrix metalloproteinases-4 and troponin I in cytokine-treated hearts. The collagen content of the heart was not affected by cytokine treatment. A neutralizing anti-MMP-2 antibody or the MMP inhibitors Ro31-9790 or PD166793 attenuated the decline in myocardial function. Moreover, the MMP-2 antibody prevented the decline in myocardial MMP-2 and troponin I levels. CONCLUSIONS: Myocardial contractile dysfunction caused by pro-inflammatory cytokines results in MMP-2 activation and a decline in tissue inhibitor of matrix metalloproteinases-4 in the heart. Troponin I is also a target for the proteolytic action of MMP-2 during acute heart failure triggered by pro inflammatory cytokines. Inhibition of MMPs may be a novel pharmacological strategy for the treatment of acute inflammatory heart disease. PMID- 12566116 TI - Increased expression of cardiac angiotensin II type 1 (AT(1)) receptors decreases myocardial microvessel density after experimental myocardial infarction. AB - OBJECTIVE: To study the effects of increased levels of myocardial angiotensin II type 1 (AT(1)) receptor on microvascular growth following myocardial infarction (MI). METHODS: MI was created in transgenic rats (TGR) with a cardioselective overexpression of the AT(1) receptor. We used Sprague-Dawley (SD) rats as controls. Some of the rats were treated with the selective AT(1) receptor blocker losartan (Los). Rats were sacrificed after 3 weeks. RESULTS: MI caused left ventricular (LV) hypertrophy and LV dysfunction in both SD and TGR, which was prevented by AT(1) receptor blockade. Furthermore, MI decreased microvessel density in the non-infarcted myocardium (SD MI: 1653+/-37/mm(2), P<0.01 vs. sham operated controls), however, microvessel density decreased significantly more in TGR with MI (1298+/-33/mm(2), P<0.01 vs. SD MI). AT(1) receptor blockade restored microvessel density (SD MI Los: 2046+/-195/mm(2); TGR MI Los: 1742+/-47/mm(2); P<0.01 vs. untreated). The differences in microvessel density were still present after correction for LV hypertrophy. The increase in microvessel density after AT(1) receptor blockade was not accompanied by increased myocardial vascular endothelial growth factor (VEGF) levels. Microvessel density correlated with parameters of myocardial stretch, such as LV end-diastolic pressure (-0.681, P<0.001) and N-ANP (-0.424, P=0.01). CONCLUSIONS: Microvessel density after MI is decreased when the AT(1) receptor is overexpressed, and this is amenable to AT(1) receptor blockade. This suggests that efficacy of AT(1) receptor blockers post-MI may not only be due to attenuation of LV remodeling, but also to a stimulatory effect on angiogenesis. PMID- 12566117 TI - Gene expression profiling of alpha(1b)-adrenergic receptor-induced cardiac hypertrophy by oligonucleotide arrays. AB - OBJECTIVE: Cardiac hypertrophy is closely associated with the development of cardiomyopathies that lead to heart failure. The alpha(1B) adrenergic receptor (alpha(1)-AR) is an important regulator of the hypertrophic process. Cardiac hypertrophy induced by systemic overexpression of the alpha(1b)-AR in a mouse model does not progress to heart failure. We wanted to explore potential gene expression differences that characterize this type of hypertrophy that may identify genes that prevent progression to heart failure. METHODS: Transgenic and normal mice (B6CBA) representing two time points were compared; one at 2-3 months of age before disease manifests and the other at 12 months when the hypertrophy is significant. Age-matched hearts were removed, cRNA prepared and biotinylated. Aliquots of the cRNA was subjected to hybridization with Affymetrix chips representing 12,656 murine genes. Gene expression profiles were compared with normal age-matched controls as the baseline and confirmed by Northern and Western analysis. RESULTS: The non-EST genes could be grouped into five functional classifications: embryonic, proliferative, inflammatory, cardiac-related, and apoptotic. Growth response genes involved primarily Src-related receptors and signaling pathways. Transgenic hearts also had a 60% higher Src protein content. There was an inflammatory response that was verified by an increase in IgG and kappa-chained immunoglobulins by western analysis. Apoptosis may be regulated by cell cycle arrest through a p53-dependent mechanism. Cardiac gene expression was decreased for common hypertrophy-inducing proteins such as actin, collagen and GP130 pathways. CONCLUSIONS: Our results suggest a profile of gene expression in a case of atypical cardiac hypertrophy that does not progress to heart failure. Since many of these altered gene expressions have not been linked to heart failure models, our findings may provide a novel insight into the particular role that the alpha(1B)AR plays in its overall progression or regression. PMID- 12566118 TI - Differential regulation of nitric oxide synthases and their allosteric regulators in heart and vessels of hypertensive rats. AB - OBJECTIVE: Nitric oxide synthase (NOS)-derived nitric oxide (NO) production is regulated posttranslationally through enzyme's inhibitory interaction with the caveolar coat protein, caveolin and stimulatory interaction with the chaperone heat shock protein, Hsp90. However, changes in the expression of these regulators with the development of hypertrophic cardiomyopathy are unknown. METHODS: Histochemical and immunoblotted signals for the NOS isoforms, caveolin and Hsp90 were compared in left ventricle (LV) and aortic or mesenteric vessels between spontaneously hypertensive rats (SHR; 18 and 63 weeks old) and age-matched normotensive Wistar-Kyoto (WKY) rats. To assess functional impacts on downstream NO signaling, superoxide anions (O(2)(-)) and cGMP contents were measured in the same tissues by oxidative fluorescent hydroethidine staining and enzyme immunoassay, respectively. RESULTS: Compared with levels in age-matched WKY rats, endothelial NOS (eNOS) proteins were increased in aorta of SHR at 18 weeks. Conversely, aortic caveolin-1 and -3 were decreased in SHR, whereas Hsp90 remained unchanged. In LV tissue of SHR at 18 weeks, caveolin-1 and -3 were similarly decreased, but Hsp90 upregulated, together with a downregulation of eNOS. However, at 63 weeks, both eNOS and neuronal NOS (nNOS) were markedly upregulated in the LV of SHR, together with an upregulation of Hsp90. No difference in cardiac and aortic cGMP contents was found between the two strains. In LV sections, O(2)(-) generation was higher in older compared with younger rats from both strains and highest in 63 weeks SHR. CONCLUSIONS: Changes in NOS protein abundance in SHR rats compared with WKY controls are differentially regulated according to the age of hypertension and the tissue examined and are not necessarily correlated with cGMP contents. The coordinate expressional changes in NOS isoforms and their allosteric regulators, such as caveolin and Hsp90, may act as a compensatory mechanism to maintain the production of bioactive NO in the face of increased oxidant stress. PMID- 12566119 TI - Differential sensitivity of atrial and ventricular K(ATP) channels to metabolic inhibition. AB - OBJECTIVE: The aim is to compare the activation of ATP-sensitive potassium channels (K(ATP) channels) in intact and metabolically impaired atrial and ventricular myocytes. METHODS: The K(ATP) channel current is measured by whole cell and gramicidin-perforated patch clamp recordings in 164 cultured neonate rat cardiomyocytes. RESULTS: In whole cell recordings with 84 micromol/l ADP in pipette, spontaneous activity is significantly higher in atrium than ventricle, and EC(50) for the K(ATP) channel opener diazoxide is 0.13 micromol/l (atrium) versus 3.1 micromol/l (ventricle). With an ATP-regenerating system in pipette, EC(50) for diazoxide is 19.7 micromol/l (atrium) versus 54.9 micromol/l (ventricle). In gramicidin-perforated patch recordings, atrial myocytes respond significantly to 100 nmol/l of the mitochondrial protonophore CCCP, while ventricular myocytes do not. EC(50) for diazoxide is 129 micromol/l (atrium) versus >2500 micromol/l (ventricle) for myocytes exposed to CCCP, and 676 versus >2500 micromol/l, respectively, without CCCP. CONCLUSIONS: (1) K(ATP) channels are significantly more sensitive to metabolic inhibition in atrial than ventricular myocytes. (2) Sensitivity of atrium versus ventricle to the channel opener diazoxide increases from 3:1 to > or = 24:1 with ADP or metabolic inhibition. If extended to intact hearts, the results would predict a higher atrial sensitivity to ischemia, and a high sensitivity of the ischemic atrium to K(ATP) channel openers. PMID- 12566120 TI - Genetic basis for chamber-specific ventricular phenotypes in the rat infarct model. AB - BACKGROUND: We, and others, have previously reported a strong correlation between increased inter-ventricular dispersion of repolarization and the occurrence of fatal arrhythmia in animal models of CHF. The existence of this and other such distinct electrophysiologic phenotypes in right (RV) vs. left ventricles (LV) could be explained by chamber-specific patterns of gene expression. METHODS: We employed microarray gene profiling of 13824 sequence-verified, nonredundant rodent cDNAs to compare myocardial gene expression in RV vs. LV of rats with surgically induced myocardial infarction (MI: n=3) and in sham-operated animals (Sham: n=3). RESULTS: Significant LV infarction (32+/-4% LV) and severe CHF were observed in all MI animals at 4 weeks. In Sham animals, 937 genes exhibited significant differential expression in RV vs. LV myocardium. In MI animals, 1158 genes exhibited significant differential expression in RV vs. LV. Of those genes exhibiting significant differential expression, only 241 were common to both Sham and MI animals. Differentially expressed genes included those involved in signal transduction, cell growth and maintenance, and apoptosis. Genes with potential roles in altered dispersion of repolarization included voltage-dependent Ca(2+) channel gamma subunit (MI 8-fold increasing) and K(+) inwardly rectifying channel subfamily J, member 10 (MI 6-fold decreasing). Gap junction membrane channel protein alpha 4 (MI 6-fold decreasing) and cardiac troponin I (MI 8-fold decreasing) were also significantly differentially expressed. Inter-ventricular comparisons revealed significantly greater alterations in gene expression vs. intra-ventricular comparisons. CONCLUSIONS: Microarray gene profiling has revealed candidate genes, some of them novel, which may account for chamber specific ventricular electrophysiologic phenotypes, both in physiologic as well as in arrhythmogenic states such as CHF. PMID- 12566121 TI - Coordinated down-regulation of KCNQ1 and KCNE1 expression contributes to reduction of I(Ks) in canine hypertrophied hearts. AB - OBJECTIVE: In animal models of hypertrophy, electrical remodeling giving rise to QT prolongation occurs rapidly and is associated with the development of torsade de pointes (TdP) arrhythmias and sudden death. Chronic AV block (CAVB)-induced hypertrophy in dogs has been associated with a reduction in the slow component (I(Ks)) of the delayed rectifier potassium current (I(K)), which contributes to a prolongation of ventricular repolarization, the development of an acquired form of long QT, and the substrate for triggered activity and TdP. The present study was designed to probe the molecular basis for the decrease in I(Ks) by studying the characteristics of KCNE1 and KCNQ1, the putative genes responsible for formation of the channel. METHODS AND RESULTS: Using a combination of Northern blot, competitive multiplex PCR and immunoblot assays, we found that CAVB reduces KCNE1 and KCNQ1 RNA in the canine ventricles by 70 and 80%, respectively. Protein levels of KCNE1 and KCNQ1 were reduced by 60 and 50%, respectively. We also demonstrate at the molecular level the basis for inter-ventricular difference in I(Ks) density previously reported in hearts of normal dogs and show the basis for reduction of this difference in the CAVB dog. CONCLUSIONS: Our results indicate that the CAVB-induced reduction in I(Ks) is due to a down-regulation of KCNE1 and KCNQ1 transcription. The data suggest that electrical remodeling of the cardiac ventricle during hypertrophy involves regulation of the gene expression through modulation of transcriptional and translational regulatory pathways. The reduction in KCNE1 and KCNQ1 expression increases the dependence of ventricular repolarization on the rapid component of I(K) and may potentiate the action of Class III antiarrhythmic agents. PMID- 12566122 TI - Modulation of the hyperpolarization-activated current (I(f)) by calcium and calmodulin in the guinea-pig sino-atrial node. AB - The aim of this study was to investigate possible regulation of the hyperpolarization-activated current (I(f)) by cytosolic calcium in guinea-pig sino-atrial (SA) node cells. Isolated SA node cells were superfused with physiological saline solution (36 degrees C) and the perforated patch voltage clamp technique used to record I(f) activated by hyperpolarizing voltage steps. A 10-min loading of SA node cells with the calcium chelator BAPTA (using 10 microM BAPTA-AM) significantly reduced the amplitude of I(f) at all potentials studied (69+/-8% at -80 mV, n=6). BAPTA loading also shifted the voltage of half activation (V(h)) of the conductance from -83+/-2 mV in control to -93+/-2 mV in BAPTA (n=6) without significantly altering the slope of activation. The calmodulin antagonists W-7 (10 microM), calmidazolium (25 microM) and ophiobolin A (20 microM) caused similar reductions in I(f) amplitude (73+/-4, 86+/-9 and 59+/-6% at -80 mV, n=6, 5 and 4, respectively) and shifts in V(h) (11+/-3, 14+/-3 and 8+/-2 mV). In cells pre-treated with W-7, exposure to BAPTA caused no further reduction in current amplitude (n=6). I(f) current amplitude was unaffected by the calmodulin dependent kinase (CaMKII) inhibitor KN-93 (1 microM) although this CaMKII inhibition did reduce L-type calcium by 48+/-19% at 0 mV (n=3). These results are consistent with a role for calcium and calmodulin in the regulation of I(f), via a mechanism that is independent of CaMKII. Alterations in intracellular calcium during the cardiac cycle may be involved in fine tuning the voltage-dependent properties of I(f) and may thus determine its relative contribution to pacemaking in the SA node. PMID- 12566123 TI - The effect of myosin light chain 2 dephosphorylation on Ca2+ -sensitivity of force is enhanced in failing human hearts. AB - OBJECTIVE: Phosphorylation of the myosin light chain 2 (MLC-2) isoform expressed as a percentage of total MLC-2 was decreased in failing (21.1+/-2.0%) compared to donor (31.9+/-4.8%) hearts. To assess the functional implications of this change, we compared the effects of MLC-2 dephosphorylation on force development in failing and non-failing (donor) human hearts. METHODS: Cooperative effects in isometric force and rate of force redevelopment (K(tr)) were studied in single Triton-skinned human cardiomyocytes at various [Ca(2+)] before and after protein phosphatase-1 (PP-1) incubation. RESULTS: Maximum force and K(tr) values did not differ between failing and donor hearts, but Ca(2+)-sensitivity of force (pCa(50)) was significantly higher in failing myocardium (Deltap Ca(50)=0.17). K(tr) decreased with decreasing [Ca(2+)], although this decrease was less in failing than in donor hearts. Incubation of the myocytes with PP-1 (0.5 U/ml; 60 min) decreased pCa(50) to a larger extent in failing (0.20 pCa units) than in donor cardiomyocytes (0.10 pCa units). A decrease in absolute K(tr) values was found after PP-1 in failing and donor myocytes, while the shape of the K(tr) Ca(2+) relationships remained unaltered. CONCLUSIONS: Surprisingly, the contractile response to MLC-2 dephosphorylation is enhanced in failing hearts, despite the reduced level of basal MLC-2 phosphorylation. The enhanced response to MLC-2 dephosphorylation in failing myocytes might result from differences in basal phosphorylation of other thin and thick filament proteins between donor and failing hearts. Regulation of Ca(2+)-sensitivity via MLC-2 phosphorylation may be a potential compensatory mechanism to reverse the detrimental effects of increased Ca(2+)-sensitivity and impaired Ca(2+)-handling on diastolic function in human heart failure. PMID- 12566124 TI - Antihypertrophic actions of the natriuretic peptides in adult rat cardiomyocytes: importance of cyclic GMP. AB - Atrial natriuretic peptide (ANP) prevents hypertrophy of neonatal cardiomyocytes. However, whether this effect is retained in the adult phenotype or if other members of the natriuretic peptide family exhibit similar antihypertrophic properties, has not been elucidated. OBJECTIVE: Our objective was to examine whether the natriuretic peptides protect against adult cardiomyocyte hypertrophy in vitro. METHODS: Adult rat cardiomyocytes were incubated with angiotensin II (Ang II)+/-ANP, B-type (BNP) or C-type (CNP) natriuretic peptides for determination of [3H]phenylalanine incorporation, c-fos mRNA expression and cyclic GMP. The effects of 8-bromo-cyclic GMP (cyclic GMP analogue), HS-142-1 (particulate guanylyl cyclase inhibitor) and KT5823 (cyclic GMP-dependent protein kinase inhibitor) were also investigated. RESULTS: Ang II-stimulated increases in markers of hypertrophy, [3H]phenylalanine incorporation (to 136+/-3% of control, n=9) and c-fos mRNA expression (4.3+/-1.4-fold, n=5), were completely prevented by each of ANP, BNP or CNP. This protective action was accompanied by increased cardiomyocyte cyclic GMP. Inhibitory actions on [3H]phenylalanine incorporation were mimicked by 8-bromo-cyclic GMP, and were abolished by HS-142-1. KT5823 blocked the response to BNP and CNP, but not to ANP. CONCLUSION: ANP prevents hypertrophy of adult rat cardiomyocytes. This protective action is shared by BNP and CNP and involves activation of particulate guanylyl cyclase receptors. Antihypertrophic effects of BNP and CNP are mediated through cyclic GMP-dependent protein kinase, but ANP can activate additional pathways independent of cyclic GMP to prevent adult cardiomyocte hypertrophy. These novel findings are of interest particularly since BNP appears to exert antifibrotic rather than antihypertrophic actions in vivo, while CNP is thought to act at least in part via the endothelium. PMID- 12566125 TI - Cardiac overexpression of monocyte chemoattractant protein-1 in transgenic mice mimics ischemic preconditioning through SAPK/JNK1/2 activation. AB - OBJECTIVE AND METHODS: Although a beneficial association between innate immunity and ischemic preconditioning has recently been proposed, the mechanisms responsible for this link are poorly understood. To test the hypothesis that pro inflammatory cytokines have a beneficial role in the activation of the cell survival pathway mediated by ischemic preconditioning, we have studied transgenic mice with cardiac myocyte specific overexpression of murine monocyte chemoattractant protein-1 (MCP-1). The resistance to ischemia was studied by performing 45-min (with or without injection of the SAPK/JNKs inhibitor D-JNKI1) and 3-day left coronary artery occlusions as well as 45-min left coronary artery occlusion followed by 3 days of reperfusion. In addition, quantitative Western blot analyses for TNF-alpha, and SAPK/JNK1/2, ERK1/2 and p38 activity were performed. RESULTS: Infarct size, expressed in percent of either the risk area or the left ventricle, was reduced in transgenic mice when compared with control after both, 45-min (14.7+/-2.6% vs. 52.0+/-2.4%; P<0.05) and 45-min occlusion followed by 3 days of reperfusion (23.2+/-1.8% vs. 30.0+/-1.8%; P<0.05) but it was not significantly different for 3-day occlusion. Western blot analyses showed significantly increased levels of TNF-alpha (1.8-fold) and phosphorylated SAPK/JNK1/2 (1.5-fold) in transgenic hearts. Phosphorylated-ERK1/2, and phosphorylated-p38 levels were unchanged. Immunohistochemistry revealed that in transgenic mice monocytes/macrophages, lymphocytes, and fibroblasts are the source of TNF-alpha, whereas myocytes have increased phosphorylated-SAPK/JNK1/2 levels. In addition, injection of the SAPK/JNKs inhibitor D-JNKI1 partially abrogated the cardioprotective effect observed in untreated transgenic mice. CONCLUSION: Overexpression of MCP-1 by cardiomyocytes causes chronic infiltration and activation of leukocytes, resulting in elevated TNF-alpha secretion and SAPK/JNK1/2 activation. The activation of this pathway is in part responsible for the preconditioning effect of MCP-1 overexpression. These results show a possible beneficial link between innate immunity and ischemic preconditioning through MAP kinase activation. PMID- 12566126 TI - Acute vascular effects of the selective estrogen receptor modulator EM-652 (SCH 57068) in the rat mesenteric vascular bed. AB - OBJECTIVE: Selective estrogen receptor modulators (SERMs) represent a class of compounds that act as either estrogen receptor agonist or antagonist in a tissue selective manner. SERMs exert beneficial effects on bone and lipids but are not associated with an increased risk of breast or uterine carcinoma. 17beta estradiol (E2) and SERMs such as raloxifene and tamoxifen acutely relax coronary arteries. EM-652 (SCH 57068) is a 4th generation SERM acting as pure antiestrogen in the mammary gland and endometrium. The effects of SERMs on the mesenteric vasculature are unknown. In the present study, the vascular effects of EM-652 and E2 on the rat mesentery were investigated. METHODS: Isolated perfused (5 ml/min) mesenteric vascular bed (MVB) was preconstricted with methoxamine. Increasing doses (0.1-10 microM) of EM-652 or E2 were infused into the perfusate. RESULTS: EM-652 and E2 relaxed MVBs removed from intact and gonadectomized female and male rats. The amplitude of EM-652 responses was consistently greater than those of E2 and its potency was similar or greater than that of other SERMs. EM-652 and E2 relaxed MVB by an endothelium-independent mechanism. The estrogen receptor (ER) antagonist ICI 182,780 attenuated E2-induced relaxations but only partially block the effects of EM-652. Inhibition of the nitric oxide synthase/cGMP pathway with N(G)-nitro-arginine-methyl-ester (L-NAME) and 1H-(1,2,4)oxadiazolo(4,3 a)quinoxaline-1-one (ODQ) or of prostanoid synthesis with indomethacin failed to reduce EM-652 responses. The vascular effects of EM-652 were also unaffected by potassium channels blockers or inhibitors/scavengers of reactive oxygen species. EM-652 attenuated the vasoconstrictor responses induced by adrenergic agonists and endothelin-1. CONCLUSIONS: EM-652 acutely relaxes the mesenteric vasculature by an endothelium-independent pathway which is partly mediated by ER, providing a novel mechanism by which this SERM may exert beneficial actions on the vascular system. PMID- 12566127 TI - Effects of local all-trans-retinoic acid delivery on experimental atherosclerosis in the rabbit carotid artery. AB - BACKGROUND: Retinoids regulate a variety of biological processes and play an important role in cell differentiation and proliferation. All-trans retinoid acid (atRA) is known to inhibit smooth muscle cell growth and thus is supposed to have favorable effects on the incidence of restenosis after percutaneous coronary interventions. The broad biological spectrum, however, leads to numerous severe side effects which limit the clinical use of a systemic application of atRA. In order to avoid systemic side effects, local delivery of atRA is preferable. The aim of this study was to evaluate the effects of atRA on the response to injury in a second-injury model of experimental balloon angioplasty. METHODS: After induction of a fibromuscular plaque in the right carotid artery of 40 New Zealand rabbits, 35 animals underwent balloon angioplasty of the preformed plaque formation. Subsequent local atRA delivery (10 ml, 10 microM) with the double balloon catheter was performed in 15 animals. Five animals received vehicle only as sham controls, and five animals were solely electrostimulated, 15 animals served as control group with balloon angioplasty only. Vessels were excised 7 days (n=15) and 28 days (n=30) after intervention. Immunocytochemistry with antibodies against smooth muscle alpha-actin and myosin, bromodeoxyuridine, macrophages, collagen I and III and von Willebrand factor was performed. Quantitative analysis was done by computerized morphometry. RESULTS: After local atRA delivery in vivo, the extent of stenosis was markedly reduced with 21.7+/ 8.3% (mean+/-S.D.) 4 weeks after intervention compared to 31.8+/-13.4% in balloon dilated animals (P=0.0937). Both a reduced early neointimal proliferation (P=0.0002) and an increase in overall vessel diameter (4 weeks after intervention, P=0.0264) contributed to a limitation of restenosis in atRA-treated animals. Immunocytochemistry revealed a more intense alpha-actin staining pattern after local atRA therapy indicating redifferentiating effects of atRA on vascular smooth muscle cells. CONCLUSIONS: Local delivery of atRA led to limitation of restenosis formation in this animal model. The concept of a local atRA therapy might be a promising way to exploit the potential of atRA for vascular indications while minimizing the severe side effects of systemic retinoid therapy. PMID- 12566128 TI - The alpha 1-adrenoceptor profile in human skeletal muscle resistance arteries in critical limb ischaemia. AB - OBJECTIVE: We recently reported the hyper-responsiveness of human skeletal muscle resistance arteries (SkMRAs) to noradrenaline in critical limb ischaemia (CLI). In this study we investigated the characteristics of alpha(1)-adrenoceptor subtypes and evaluated the agonist affinity and adrenoceptor reserve in the ischaemic arteries. METHODS: Human SkMRAs were isolated from non-ischaemic and ischaemic areas of limbs amputated for CLI. Subcutaneous resistance arteries were isolated from inguinal biopsies from healthy subjects. Arterial segments were mounted on a small vessel wire myograph. RESULTS: Contractile responses to agonists, adrenaline and A-61603 (alpha(1A)-selective) were significantly increased in ischaemic arteries compared to those in non-ischaemic arteries. Receptor inactivation studies indicated an increase in the alpha-adrenoceptor reserve in the ischaemic arteries but the affinity of noradrenaline was unaffected. Healthy subcutaneous arteries had a similar noradrenaline affinity but a higher receptor reserve than skeletal muscle arteries. In the ischaemic arteries, the antagonists prazosin (alpha(1)-selective), 5-methyl-urapidil (alpha(1A)-selective) and BMY 7378 (alpha(1D)-selective) produced rightward shifts in the concentration response curves (CRCs) of noradrenaline giving pK(B)s of 9.6+/-0.3, 8.4+/-0.2 and 7.1+/-0.4, respectively. Pretreatment with 10 microM chloroethylclonidine decreased the contractile responses to noradrenaline and A 61603 to 57+/-7 and 72+/-4% of their respective controls. CONCLUSIONS: These results demonstrate that the ischaemic SkMRAs have an increased alpha adrenoceptor reserve with no change in the predominant alpha(1A)-adrenoceptor profile. PMID- 12566129 TI - Vitamin E inhibits lipid peroxidation-induced adhesion molecule expression in endothelial cells and decreases soluble cell adhesion molecules in healthy subjects. AB - OBJECTIVE: In a combination of in vivo and in vitro studies, we investigated mechanisms via which alpha-tocopherol, a lipid soluble form of vitamin E, can directly affect endothelial activation as induced by H(2)O(2) and TNFalpha. METHODS: We measured effects of alpha-tocopherol on H(2)O(2)-induced lipid peroxidation as determined with a fluorescent C-11 BODIPY(581/591) probe and on adhesion molecule expression in cultured endothelial cells. In 20 healthy volunteers treated with increasing doses of alpha-tocopherol up to 800 IU/ml for 12 weeks, plasma levels of soluble cell adhesion molecules (sCAMs) and C-reactive protein were measured. RESULTS: We showed that alpha-tocopherol protects cultured endothelial cells against H(2)O(2)-induced lipid peroxidation, while TNFalpha did not induce lipid peroxidation. Moreover, alpha-tocopherol attenuated H(2)O(2)-, but not TNFalpha-induced increases in adhesion molecule expression. In healthy persons, alpha-tocopherol decreased plasma levels of sE-selectin from 65+/-6 to 60+/-6 ng/ml (P=0.002), sVCAM from 893+/-31 to 853+/-23 ng/ml (P=0.022), and sICAM from 483+/-21 to 463+/-16 ng/ml (P=0.048). C-Reactive protein, as a sensitive marker of low grade inflammation, was not significantly affected. CONCLUSION: alpha-Tocopherol specifically inhibits lipid peroxidation-induced endothelial activation in vitro. The observed vitamin E-induced decrease in sCAMs in control subjects suggests that lipid peroxidation can take place in healthy individuals. Although vitamin E supplementation may be especially effective in specific groups of patients exposed to increased oxidative stress, our study suggests that vitamin E supplementation can be of benefit in healthy individuals as well. PMID- 12566130 TI - Overexpression of matrix metalloproteinase-9 promotes intravascular thrombus formation in porcine coronary arteries in vivo. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) cause extracellular matrix degradation and may be involved in the rupture of atherosclerotic plaques by degrading fibrous cap, resulting in the intravascular thrombus formation. Here we examined whether local overexpression of MMP-9 alters the characteristics of arteriosclerotic vascular lesions and promotes thrombosis after balloon injury in porcine coronary arteries in vivo. METHODS AND RESULTS: Balloon angioplasty was performed in the left coronary arteries followed by injection of adenovirus vector solution encoding either MMP-9 or beta-galactosidase (beta-gal) gene into the injured coronary arteries. Three weeks after the gene transfer, histological examination demonstrated that macroscopic intravascular thrombus formation was noted at the MMP-9-transfected site but not at the beta-gal-transfected site. Microscopic intramural thrombus area was significantly larger at the MMP-9 transfected site as compared to the beta-gal-transfected site. Co-transfection of tissue inhibitor of metalloproteinase-1 (TIMP-1) with MMP-9 prevented the intravascular thrombus formation in vivo. Western blot analysis revealed the reduced expression of intact tissue factor pathway inhibitor-1 and the increased tissue factor (TF) expression at the MMP-9-transfected sites. CONCLUSION: These results provide the first in vivo evidence that overexpression of MMP-9 promotes intravascular thrombus formation after balloon injury due in part to the activation of TF-mediated coagulation cascade. PMID- 12566131 TI - Dietary fat, the Mediterranean diet, and health: reports from scientific exchanges, 1998 and 2000. Introduction. PMID- 12566132 TI - Dietary fat consensus statements. PMID- 12566133 TI - Epidemiologic studies on dietary fats and coronary heart disease. AB - The results of large prospective epidemiologic investigations support the hypothesis that coronary disease risk depends on the quality rather than quantity of dietary fat. Whereas saturated fat and cholesterol appear to increase the risk of coronary heart disease (CHD) as predicted by their effects on blood lipids, strong evidence has emerged that the deleterious effects of trans unsaturated fatty acids (trans fatty acids) extend beyond those predicted by their well-known adverse influence on the ratio of low-density lipoprotein to high-density lipoprotein cholesterol. On the other hand, increased consumption of the polyunsaturated fats, linoleic acid and linolenic acid, appears to reduce the risk of CHD. PMID- 12566134 TI - Randomized clinical trials on the effects of dietary fat and carbohydrate on plasma lipoproteins and cardiovascular disease. AB - Several dietary approaches have reduced cardiovascular events in randomized clinical trials. Replacing saturated fat with polyunsaturated fat prevented coronary events in men, and a Mediterranean diet and fatty fish improved survival. None of these trials had much impact on total fat intake but rather increased vegetable oils, n-3 fatty acids, or many other plant foods or nutrients that are linked to coronary prevention. The reductions in cardiovascular disease (CVD) caused by these dietary therapies compare favorably with drug treatments for hyperlipidemia and hypertension. Improvement in blood lipid risk factors is an important mechanism to explain the results of trials of unsaturated fats. When saturated or trans unsaturated fats are replaced with monounsaturated or n-6 polyunsaturated fats from vegetable oils, primarily low-density lipoprotein (LDL) cholesterol decreases. The LDL to high-density lipoprotein (HDL) cholesterol ratio decreases. When carbohydrates are used to replace saturated fats, in a low fat diet, LDL and HDL decrease similarly, and the ratio is not improved; triglycerides increase as well when carbohydrate increases, except when low glycemic index foods are used. The n-3 polyunsaturated fats in fish oils suppress cardiac arrhythmias and reduce triglycerides, but they have little effect on LDL or HDL cholesterol levels. The theme should be that diet has benefits that come directly from foods, as well as from the reduction in saturated fats, cholesterol, meats, and fatty dairy foods. It is likely that many diets could be designed that could prevent CVD. This potential diversity is crucial for engaging the diverse cultures and tastes of people worldwide in cardiovascular disease prevention. PMID- 12566135 TI - Diet composition and the metabolic syndrome: what is the optimal fat intake? AB - Two cholesterol-raising fatty acids in the diet, saturated fatty acids and trans fatty acids, increase the serum low-density lipoprotein cholesterol concentration. This fact justifies the recommendation of a reduced intake of cholesterol-raising fatty acids. Emerging data suggest that diets higher in unsaturated fatty acids, particularly monounsaturated fatty acids, have several advantages over high-carbohydrate intakes. This advantage appears to hold, particularly for populations having a high prevalence of insulin resistance, such as the US population. If the US public were to modify its eating habits in the direction of better weight control and more exercise, higher intakes of carbohydrate might be better tolerated. At the same time, the experience with the Mediterranean population reveals that in healthier populations, diets relatively high in unsaturated fatty acids are well tolerated and are associated with a low prevalence of both coronary heart disease and type 2 diabetes. PMID- 12566136 TI - High-complex carbohydrate or lente carbohydrate foods? AB - Current dietary guidelines of the American Diabetes Association emphasize the importance of minimizing risk factors for cardiovascular disease while maximizing diabetes control. Potential advantages are seen for increased monounsaturated fat intake, but only the quantity rather than the quality of the carbohydrate is considered important. However, review of the carbohydrate issue suggests that many cultures now at high risk of diabetes originally consumed starchy staples higher in fiber and with a lower glycemic index than eaten currently. Furthermore, diets high in cereal fiber have been associated with improved glycemic control, and low glycemic index diets resulted in reduction in glycosylated proteins in type 1 and 2 diabetes. Finally, large cohort studies have demonstrated beneficial effects of cereal fiber and low glycemic index carbohydrate foods in reducing the risk both for diabetes and cardiovascular disease. The effect of insoluble cereal fiber is not readily explained, but a low glycemic index may result from a slower rate of carbohydrate absorption. Increased meal frequency as a model for reducing the rate of carbohydrate absorption has been shown to reduce day-long glucose and insulin levels in type 2 diabetes and reduce serum lipids in nondiabetic subjects. Therefore, there appears to be a potential role for low glycemic index, high-cereal fiber foods for prevention and treatment of diabetes. Both the nature of the dietary fat and the carbohydrate should be considered as potentially modifiable factors that together with weight control and exercise may play a role in diabetes and cardiovascular disease prevention and treatment. PMID- 12566137 TI - Dietary fat and diabetes: a consensus view. AB - There may be no area in nutrition more controversial or subject to more changes in policy than dietary fat recommendations for individuals with diabetes. At one extreme, diets very low in carbohydrates with very high levels of fat were advocated in the early days of diabetes therapy. At the other extreme, the literature in the late 1980s and early 1990s emphasized diets very low in fat and high in carbohydrates. This conference has shown that too much information is missing for a definitive recommendation to be made concerning diet composition for diabetic individuals, but it also has led to the realization that we may now have considerable flexibility in our dietary recommendations for individuals with diabetes. PMID- 12566138 TI - Low-fat diets are preferred. AB - This short review summarizes 4 main reasons for which low-fat diets are preferred to limit excessive weight gain: (1) For metabolic reasons, fat intake does not measurably stimulate fat oxidation; dietary fat above energy requirements is stored in adipose tissue. (2) Diets that are high in fat or are energy dense have a weak satiating effect and promote a passive overconsumption of energy relative to need. (3) A recent meta-analysis on the effect on body weight loss of low-fat diets followed for >2 months showed a significant weight difference of 3.3 kg between the diet and the control groups. A low-fat diet may also be beneficial in helping maintain weight loss. (4) Low-fat diets are also advocated to lower the risk of coronary heart disease and certain forms of cancer. There is no evidence showing that the small physiologic reduction of plasma highdensity lipoprotein cholesterol levels with a low-fat diet is detrimental. PMID- 12566139 TI - Dietary fat is not a major determinant of body fat. AB - The percentage of energy from fat in diets has been thought to be an important determinant of body fat, and several mechanisms have been proposed. Comparisons of diets and the prevalence of obesity between affluent and poor countries have been used to support this relationship, but these contrasts are seriously confounded by differences in physical activity and food availability. Within areas of similar economic development, regional intake of fat and prevalence of obesity have not been positively correlated. Randomized trials are the preferable method to evaluate the effect of dietary fat on adiposity and are feasible because the number of subjects needed is not large. In short-term trials, a modest reduction in body weight is typically seen in individuals randomized to diets with a lower percentage of calories from fat. However, compensatory mechanisms appear to operate, because in randomized trials lasting >or=1 year, fat consumption within the range of 18% to 40% of energy appears to have little if any effect on body fatness. The weighted mean difference was -0.25 kg overall and +1.8 kg (i.e., less weight loss on the low-fat diets) for trials with a control group that received a comparable intensity intervention. Moreover, within the United States, a substantial decline in the percentage of energy from fat during the last 2 decades has corresponded with a massive increase in the prevalence of obesity. Diets high in fat do not appear to be the primary cause of the high prevalence of excess body fat in our society, and reductions in fat will not be a solution. PMID- 12566140 TI - Consensus view on the role of dietary fat and obesity. PMID- 12566141 TI - Dietary fat and cancer. AB - Based on current epidemiologic knowledge, public health recommendations to decrease total fat intake for the prevention of cancer appear largely unwarranted. Recommendations to decrease red meat intake, particularly processed meat or beef intake, may, on the other hand, decrease the risk of colorectal cancer and prostate cancer; it may have a beneficial effect on breast cancer as well, although the evidence is much less compelling in this regard. There appears to be no particular benefit regarding cancer prevention that would accrue from reducing fat intake from vegetable sources, and in the case of breast cancer, there is some suggestive but preliminary evidence that olive oil or other sources of monounsaturated fatty acids may modestly decrease risk. Overall, recommendations focused on controlling weight by regular physical activity and avoidance of excessive energy intake from all sources; increasing plant food intake; consuming a variety of whole grains, vegetables, and fruits; and decreasing red meat intake are likely to be more effective in decreasing risk of breast, colorectal, and prostate cancer than decreasing total fat intake. This conclusion is consistent with current recommendations for cancer prevention as promulgated by the American Cancer Society. PMID- 12566143 TI - Dietary fats, eating guides, and public policy: history, critique, and recommendations. AB - Controversies over the nutrition science of dietary fat, and equally over the advice furnished to consumers about dietary fat, have confounded US nutrition policies and eating guidance for the last 90 years. This is so despite the remarkable congruence between the first US food guides (1916) and the most recent (2000 Dietary Guidelines for Americans), both of which state that dietary fats should be consumed "moderately." The 2002 Report of the US Food and Nutrition Board (issued jointly by the United States and Canada) quantifies this by stating that healthy dietary fat should constitute "25-35 percent of calories." However, the US consumer guide, the Food Guide Pyramid (released in 1992 but based on data from the early 1980s) states that dietary fats should be consumed "sparingly," which is explained to be "a diet low in fat." This direct conflict in official dietary policies causes consumer confusion and erodes efforts of public and private health promotion efforts to stem the increasing incidence of overweight and obesity in Americans. The most successful population-wide dietary behavior modification program in US history was the food rationing program in World War II. Its successes were based equally on consensus nutrition profiles for good health and messages that communicated the rationing program effectively. The current US incidence of overweight and obesity, and the chronic diseases to which they are precursors, will be mitigated and prevented only with major changes in national dietary policies and programs based on successful experiences and models. The first step in this much-needed process is acknowledgment that current dietary guidance and education policies have been and are unsatisfactory. PMID- 12566142 TI - Bioactive compounds in foods: their role in the prevention of cardiovascular disease and cancer. AB - "Bioactive compounds" are extranutritional constituents that typically occur in small quantities in foods. They are being intensively studied to evaluate their effects on health. The impetus sparking this scientific inquiry was the result of many epidemiologic studies that have shown protective effects of plant-based diets on cardiovascular disease (CVD) and cancer. Many bioactive compounds have been discovered. These compounds vary widely in chemical structure and function and are grouped accordingly. Phenolic compounds, including their subcategory, flavonoids, are present in all plants and have been studied extensively in cereals, legumes, nuts, olive oil, vegetables, fruits, tea, and red wine. Many phenolic compounds have antioxidant properties, and some studies have demonstrated favorable effects on thrombosis and tumorogenesis and promotion. Although some epidemiologic studies have reported protective associations between flavonoids or other phenolics and CVD and cancer, other studies have not found these associations. Various phytoestrogens are present in soy, but also in flaxseed oil, whole grains, fruits, and vegetables. They have antioxidant properties, and some studies demonstrated favorable effects on other CVD risk factors, and in animal and cell culture models of cancer. However, because phytoestrogens act both as partial estrogen agonists and antagonists, their effects on cancer are likely complex. Hydroxytyrosol, one of many phenolics in olives and olive oil, is a potent antioxidant. Resveratrol, found in nuts and red wine, has antioxidant, antithrombotic, and anti-inflammatory properties, and inhibits carcinogenesis. Lycopene, a potent antioxidant carotenoid in tomatoes and other fruits, is thought to protect against prostate and other cancers, and inhibits tumor cell growth in animals. Organosulfur compounds in garlic and onions, isothiocyanates in cruciferous vegetables, and monoterpenes in citrus fruits, cherries, and herbs have anticarcinogenic actions in experimental models, as well as cardioprotective effects. In summary, numerous bioactive compounds appear to have beneficial health effects. Much scientific research needs to be conducted before we can begin to make science-based dietary recommendations. Despite this, there is sufficient evidence to recommend consuming food sources rich in bioactive compounds. From a practical perspective, this translates to recommending a diet rich in a variety of fruits, vegetables, whole grains, legumes, oils, and nuts. PMID- 12566144 TI - Excess nuclear DNA in spermatozoa of guinea fowl. AB - The proportion of spermatozoa with elongated nuclei in ejaculates from a strain of guinea fowl was estimated, subjectively, to range from approximately 1 to 6%. It was confirmed by image analysis that in an ejaculate from one male, the distribution of nuclear lengths was bimodal, with a distinct population comprising 10% of spermatozoa having a mean nuclear length that was 52% greater than that of the remaining 90%. Furthermore, the mean DNA content of the 'large nuclei' population was 1.85 times (not significantly different from twice) that of the main sperm population. The proportion of large-nuclei spermatozoa that was motile was less than that of normal sperm (31% versus 59%) and the velocity of motile spermatozoa was also less (24 microm/s versus 72 microm/s). The poor motility of the large-nuclei spermatozoa in vitro was reflected in their limited performance in vivo, since only 1.1% were found associated with the egg outer perivitelline layer. This is the first report to quantify the occurrence of, presumed, polyploid spermatozoa in a domestic bird. The incidence of such spermatozoa in commercial guinea fowl and other domestic poultry and the genesis and effects on fertility of such spermatozoa may be significant. PMID- 12566145 TI - Membrane changes during different stages of a freeze-thaw protocol for equine semen cryopreservation. AB - Many theories have been postulated concerning the possible effects of cryopreservation on spermatozoa, including suggestions the freeze-thawing process produces membranes that have greater fluidity and are more fusogenic, thus inducing changes similar to those of capacitation. The main objectives of this study were to determine at what stage of the freeze-thaw process membrane changes occur and whether evaluation with chlortetracycline (CTC) stain could predict the freezability of stallion sperm. Sperm viability and state of capacitation were simultaneously evaluated using CTC and Hoechst 33258 (H258) techniques. Membrane function was evaluated using the hypo-osmotic swelling test (HOS) and progressive motility (PM) was evaluated under light microscopy at each stage of a freeze-thaw protocol. Evaluated were raw semen; after dilution and centrifugation; after redilution and equilibration at room temperature; after cooling to 5 degrees C; after super cooling to -15 degrees C; and after thawing. The most pronounced functional damage to membranes and the greatest decrease in PM occurred in samples of all stallions after thawing (P<0.05). Cryopreservation, as evaluated by CTC/H258 staining, significantly (P<0.05) affected sperm membrane integrity after centrifugation, after redilution and equilibration at room temperature and after cooling to 5 degrees C. The HOS and H258 tests gave similar results (R values of approximately 0.75) and correlated inversely with the number of live noncapacitated sperm cells (R values of approximately -0.75). Remarkably, the subpopulation of capacitated live cells was unaffected in all freeze-thawing steps and the number of live acrosome reacted cells increased by a factor of 4. However, it was not possible to determine whether the changing CTC patterns reflect a true capacitation phenomenon or an intermediate destabilized state of the sperm cell membrane. This increase may indicate that the subpopulation of functional sperm cells capable of binding to the zona pellucida increases after freeze-thawing despite the deteriorative effect of this procedure for the entire live sperm population. PMID- 12566146 TI - Postpartum reproductive function: association with energy, metabolic and endocrine status in high yielding dairy cows. AB - This study evaluated the effect of metabolic, endocrine and energy status on onset of ovarian cycle, days open (DO), and conception at first service in 90 multiparous Holstein cows, housed at a research farm. Dry matter intake, milk yield and body weight were measured daily from Week 2 antepartum (a.p.) to Week 20 postpartum (p.p.). Milk composition was determined four times per week and milk acetone was measured weekly. Blood samples for the determination of glucose, non-esterified fatty acids, cholesterol, creatinine, albumin, urea, beta hydroxybutyrate, leptin, insulin, insulin-like growth factor-1, growth hormone, 3,5,3'-triiodothyronine (T(3)), and thyroxine (T(4)) were taken 2 weeks a.p., in Weeks 1-16, and 20 p.p. between 7:30 and 9:00 h. The onset of ovarian cycle was specified by weekly gynecological examination and by skim milk progesterone determination by radioimmunoassay (twice per week). Energy balance (EB) traits were calculated and expressed as accumulated negative EB from calving to EB equilibrium, EB nadir (EBN), rate of EB recovery after EBN (EBR), and time from calving to EBN and to EB equilibrium, respectively. The onset of ovarian cycle p.p. was not related to EB. However, a low degree of EBN and a fast EBR were associated with fewer DO, and EB at first service was positively related to conception. High plasma levels of T(3) and T(4) p.p. were associated with an early start of ovarian cycle, and high concentrations of glucose and cholesterol with a short calving to conception interval. Conception at first service was positively related to EB at first service and progesterone concentration 10-13 days after first service. In conclusion, thyroid hormones may play an important role in resumption of ovarian cyclicity p.p., and a good energy status enhances the chance of conception at first service and shortens DO. PMID- 12566147 TI - Effects of Equex from different sources on post-thaw survival, longevity and intracellular Ca2+ concentration of dog spermatozoa. AB - The aims of the present study were to compare the effects of two commercial preparations (Equex STM Paste or Equex Pasta), whose active ingredient is sodium dodecyl sulphate (SDS), added to a Tris-egg yolk-based extender, on post-thaw sperm survival and longevity, as well as on the intracellular Ca(2+) concentration of dog spermatozoa during incubation at 38 degrees C. One ejaculate was collected from each of eight dogs. Each ejaculate was centrifuged, the semen plasma discarded, and the sperm pellet rediluted with a Tris-glucose-egg yolk extender containing 3% glycerol (Ext-1) at a sperm concentration of 200 x 10(6) spermatozoa (spz)/ml. The diluted semen was divided in three aliquots of equal volume and allowed to equilibrate for 1h at 4 degrees C. After equilibration, the same volume of three different second extenders was added, respectively, to each of the three aliquots: (A) Ext-2A (same composition as Ext-1 except that it contained 7% glycerol and 1% Equex STM Paste), (B) Ext-2B (same composition as that of Ext-1 except that it contained 7% glycerol and 1% Equex Pasta), and (C) Ext-2 (CONTROL: same composition as that of Ext-1 except that it contained 7% glycerol). Semen samples were packed in 0.5 ml straws and frozen on a rack 4 cm above liquid nitrogen (LN(2)) in a styrofoam box. Thawing was at 70 degrees C for 8s. Sperm motility was evaluated after thawing and at 1 h intervals for 5h at 38 degrees C by subjective examination and by using a CASA system. Plasma membrane integrity and acrosomal status were evaluated at 1, 4 and 7h post-thaw using a triple staining procedure and flow cytometry. Intracellular Ca(2+) concentration of live spermatozoa was evaluated by flow cytometry at 1, 4 and 7h post-thaw after co-loading the sperm cells with the Ca(2+) indicators Fluo 3 AM and Fura Red AM, and with PI. Post-thaw sperm survival and longevity, as well as the quality of the sperm movement, were significantly better (P<0.005) when Ext-2A (containing Equex STM Paste) was used. There was no difference between Ext-2B (containing Equex Pasta) and Ext-2 (CONTROL). The mean intracellular Ca(2+) concentration (arbitrary units) of cryopreserved spermatozoa (range: 0.23+/-0.12 to 1.26+/-0.46) was higher than that of fresh spermatozoa (0.13+/-0.06). When using Ext-2A, the live spermatozoa frequently (P=0.012) appeared divided in two subpopulations, with high (1.26+/-0.46) and low (0.27+/-0.09) intracellular Ca(2+) content, respectively. When using Ext-2B or Ext-2, the live spermatozoa were more frequently seen in a single population with low intracellular Ca(2+) concentration (0.30+/-0.35 and 0.23+/-0.12, for Ext-2B and Ext-2, respectively). PMID- 12566148 TI - FSH priming improves oocyte maturation, but priming with FSH or hCG has no effect on subsequent embryonic development in an in vitro maturation program. AB - AIM: To determine whether maturation and subsequent blastocyst development of in vitro matured oocytes can be improved by in vivo follicle stimulating hormone (FSH) or human chorionic gonadotrophin (hCG) priming, using a mouse model. EXPERIMENTAL DESIGN: Five groups of oocytes were used: in vivo control, in vitro matured (IVM) control, IVM after 24 h in vivo priming with FSH, IVM after 48 h in vivo priming with FSH and IVM after 16 h in vivo priming with hCG. In vitro fertilization (IVF) was performed on all groups. Oocyte maturation, fertilization, blastocyst development rates and blastocyst cell numbers were assessed for all groups. RESULTS: Significant improvement in oocyte maturation was observed in the two FSH priming groups compared with the IVM control group (P<0.005 and P<0.001, respectively). There were no significant differences in fertilization between all five groups. Blastocyst development was significantly higher in the in vivo control compared to the IVM groups (P<0.001). No significant differences were observed in blastocyst cell numbers among all five groups. CONCLUSIONS: While FSH priming improves the maturation rate of IVM oocytes, FSH or hCG priming does not improve development to the blastocyst stage. PMID- 12566149 TI - Effect of leukemia inhibitory factor on bovine embryos produced in vitro under chemically defined conditions. AB - The objective of these experiments was to assess putative embryotrophic effects of leukemia inhibitory factor (LIF) on bovine preimplantation development in chemically defined media. Recombinant human LIF was added to embryo culture media at a concentration of 100 ng/ml. When added for culture of morulae LIF had no positive effect on the proportion of embryos reaching the blastocyst stage. However, LIF significantly reduced development to the blastocyst stage when added for culture of 4-cell stage embryos (P<0.05). In contrast, a positive effect was found for progression of blastocyst development. In vitro blastocyst hatching rates were significantly improved in the presence of LIF (P<0.02). Number of total cells and of inner cell mass (ICM) cells were increased in LIF-treated blastocysts. In vitro survival of frozen-thawed blastocysts was not improved by adding LIF to morula stage embryos before cryopreservation. The pregnancy rate after direct transfer of cryopreserved LIF-treated embryos was not different from that for untreated control embryos. Data indicate that addition of LIF has no major beneficial effect on bovine embryos produced in these chemically defined conditions. PMID- 12566150 TI - Effect of processing and measuring procedures on estimated sizes of bull sperm heads. AB - Reported estimates of sperm head size within a species vary considerably, partly due to procedural effects. A simple India ink method was developed that provided good contrast without inducing artifacts. Semen from five fertile bulls was smeared on replicate slides and left unfixed or fixed in Carnoys solution, with ink added for background. Other slides were fixed, and sperm were stained by the Feulgen procedure. Sperm head area was measured four ways. These were linear measurements made with the aid of an ocular micrometer and an oil immersion objective, plus three methods of measuring sperm heads projected at magnification 5,000 x. The areas of unfixed and fixed sperm heads did not differ (41.5 microm(2) versus 41.6 microm(2), respectively, P>0.05). The Feulgen-stained head area was smaller (26.2 microm(2), P<0.05). Sperm head areas calculated from ocular micrometer measurements were slightly smaller (P<0.05) than areas measured using projection. Identical results obtained by two technicians were treated as duplicates and approximately half of the variation was biological, due to source of semen. There was an interaction (P<0.05) between the sample source and fixation procedures. Thus, preparative techniques must be carefully controlled, and experiments designed to partition possible interactions between the biological material sampled and procedures used. PMID- 12566151 TI - Somatic cell nuclear transfer in domestic cat oocytes treated with IGF-I for in vitro maturation. AB - Oocyte maturation and somatic cell nuclear transfer (NT) studies conducted in the domestic cat can provide valuable insights that are relevant to the conservation of endangered species of felids. The present investigation focuses on the in vitro maturation (IVM) of domestic cat oocytes stimulated by insulin-like growth factor-I (IGF-I) and their possible use as recipient cytoplasts for somatic cell NT. In Experiment I, the effects of IGF-I on cat oocyte IVM were monitored. Cumulus-oocyte complexes (COCs) were recovered in TALP-HEPES medium following ovarian follicular aspiration and were classified under a stereomicroscope into four grades using criteria based on cumulus cell investment and the uniformity of ooplasm. The COCs were either cultured in Dulbecco's modified Eagle medium (DMEM) alone as a control group or supplemented with 100 ng/ml IGF-I. After culturing for 32-34 h, oocytes were denuded and maturation rate was evaluated by observing the extrusion of the first polar body and staining with aceto-orcein. The percentages of maturation of Grades 1 and 2 oocytes were significantly increased (P<0.05) in IGF-I supplemented medium compared with medium alone (85.8 versus 65.5 and 70.3 versus 51.8, respectively) whereas the maturation rates of Grades 3 and 4 oocytes were not different. The IVM of Grade 1 oocytes was significantly higher (P<0.05) than for all other grades in both control and experimental groups. In Experiment II, the in vitro development of cat NT embryos using cumulus cells, fetal or adult fibroblasts as donor nuclei was investigated. The IVM oocytes in medium containing IGF-I were enucleated and fused with cumulus cells, fetal or adult fibroblasts between passages 2 and 4 of culture. Reconstructed embryos were cultured and monitored every 24h for progression of development through Day 9. There was no significant difference in the percentage of fusion of NT embryos using different donor nuclei whereas the cleavage rates of NT embryos reconstructed with fetal fibroblasts and cumulus cells were significantly higher (P<0.05) than those reconstructed with adult fibroblasts (72.5 and 70.7% versus 54.8%, respectively). Development of NT embryos reconstructed with adult fibroblast to the morula stage was significantly lower (P<0.05) compared with cumulus cell or fetal fibroblast donor cells (25.8% versus 37.9 or 47.5%, respectively). However, no difference was observed in development to the blastocyst stage. These results demonstrated that IGF-I promoted the IVM of domestic cat oocytes. The enucleated IVM oocytes could be used as recipient cytoplasm for fetal and adult somatic cell nuclei resulting in the production of cloned cat embryos. PMID- 12566153 TI - The effects of GnRH administration postinsemination on serum concentrations of progesterone and pregnancy rates in dairy cattle exposed to mild summer heat stress. AB - The objective of this study was to evaluate whether administration of GnRH postinsemination would improve reproductive performance in heat-stressed dairy cattle. Estrous cycles of Holstein cows were synchronized using the OvSynch protocol and cows were artificially inseminated. Cows were then administered the following treatments: control (no GnRH; n=37), GnRH (100 microg) on Day 5 (GnRH D5; n=34), or GnRH (100 microg) on Day 11 (GnRH-D11; n=34) postinsemination. Cows were provided access to both fans and sprinklers, and environmental data was collected hourly. Rectal temperatures and blood samples were obtained from cows on Days -9, -2, 0 (AI) and on alternate days from Day 5 to Day 19 postinsemination. Blood serum was collected for the analysis of progesterone (P(4)) by RIA. In a subset of cows (n=6/treatment) ultrasonography was performed on alternate days from Day 5 to Day 19 postinsemination to assess numbers of corpora lutea (CL) and CL cross-sectional areas. Pregnancy status of cows was confirmed at Day 30 postinsemination. Environmental data indicated that cows experienced mild heat stress during the trials (mean daily THI=73-77). Serum P(4) was greater (P<0.05) after Day 9 for GnRH-D5 cows and after Day 15 for GnRH-D11 cows through Day 19 postinsemination. The number of CLs present for GnRH-D5 cows was greater (P<0.05) on Day 17 than in either the control or GnRH-D11 treatment groups. On Day 17 postinsemination, both the GnRH-D5 and GnRH-D11 cows were observed to have greater (P<0.05) total CL tissue area than control cows. The interval from insemination to when serum P(4) returned to <1 ng/ml (i.e. luteolysis and return to estrus) did not differ (P>0.10) among treatment groups. Control cows (19%) tended to exhibit lower pregnancy rates (P<0.08) compared to the GnRH-D5 and GnRH-D11 treatment groups combined (35%). In summary, the treatment of heat-stressed dairy cows with GnRH postinsemination (Day 5 or 11) results in the appearance of more CL tissue, increased serum concentrations of P(4) and a tendency toward greater pregnancy rates. PMID- 12566152 TI - Use of medroxyprogesterone acetate (MAP) in lactating Holstein cows within an Ovsynch protocol: follicular growth and hormonal patterns. AB - To evaluate the effects of incorporating medroxyprogesterone acetate (MAP) in an Ovsynch protocol, cyclic lactating dairy cows were assigned randomly to two groups (control and MAP, n=8 each). Ovsynch treatment (Day 0: GnRH, Day 7: PG, Day 9: GnRH) was initiated at random stages of the estrous cycle (control) and an intravaginal polyurethane sponge impregnated with 300mg of MAP was inserted intravaginally in the MAP group at Day 0 and removed at Day 7 of the Ovsynch protocol (MAP treatment). Ovaries were scanned daily from Day 0 until the second GnRH treatment on Day 9 and from then every 6h for 36 h. Milk samples were collected three times weekly starting 17 days before the initiation of treatment to determine the stage of the cycle at the beginning of the Ovsynch protocol. Blood samples were collected to monitor estradiol (E2), progesterone (P4), LH, and 15-ketodihydro-PGF(2alpha) (PGFM) by RIA. Response to the first GnRH treatment varied with the stage of the cycle at the time of initiation of treatment, as cows in metestrous and late diestrous did not ovulate. In cows ovulating, growth rate of the new follicle was not affected by the addition of MAP. No treatment differences were found in E2 concentrations which reached a maximum at Day 9, consistent with the maximum follicular size. At Day 7, cows with luteal concentrations of P4 had increased concentrations of PGFM, but cows with basal P4 did not show an active release of prostaglandins. There were no treatment differences in the ovulatory response to the second GnRH-induced ovulation, with 11 of the 16 cows ovulating between 16 and 32 h. The addition of MAP to the Ovsynch protocol could not mimic the normal high progesterone levels needed to prevent premature ovulations in those cows with premature CL regression. PMID- 12566154 TI - Calculated follicle deviation using segmented regression for modeling diameter differences in cattle. AB - Segmented linear regression alone or in combination with simple linear regression was evaluated as an objective method to calculate the beginning of follicle deviation by modeling the sequential (Experiment 1) and non-sequential or single point (Experiment 2) differences in diameter between the future dominant (F1) and largest subordinate (F2) follicles of Wave 1 in cattle. The segmented regression consisted of Segment 1 representing the common growth phase, Segment 2 representing the period of dominance, and a Join Point connecting the two segments and representing the end of the common growth phase and the beginning of deviation. The model was fit to the diameter differences for each heifer in Experiment 1 (n=15) and the group of heifers in Experiment 2 (n=40). The optimal Join Point value that corresponded to the maximum R(2) was designated the calculated hour (Experiment 1) or diameter of F1 (Experiment 2) at the beginning of deviation. In Experiment 1, simple linear regression was used to calculate the corresponding diameter of F1 at the beginning of deviation. Observed deviation was determined by inspection of the diameter profiles of F1 and F2 for comparison to calculated deviation. In Experiment 1, the observed method determined the beginning of deviation in 80% of the heifers, whereas, the regression method calculated deviation in 93% of the heifers including two of the three heifers in which observed deviation was not discernable (no significant difference between methods). The mean hours of deviation after wave emergence (Hour 0) and diameters of F1 at the corresponding hours were not significantly different between the observed (62 h and 8.4 mm) and calculated (61 h and 8.8 mm) methods. In Experiment 2, the diameter of F1 at the beginning of calculated deviation was 8.2 mm. The results indicated that the segmented regression model can provide an objective and more accurate alternative to estimate follicle deviation, especially when observed deviation is obscured by the complexity of follicle development in some waves. PMID- 12566155 TI - Fixed-time insemination in peripuberal, lightweight replacement beef heifers after estrus synchronization with PGF2alpha and GnRH. AB - Estrus synchronization contributes to optimizing the use of time, labor, and financial resources by shortening the calving season, in addition to increasing the uniformity of the calf crop. We determined whether acceptable pregnancy rates could be achieved after synchronization of ovulation and fixed-time artificial insemination (AI) in peripuberal replacement beef heifers using gonadotropin releasing hormone (GnRH) and PGF2alpha. Crossbred heifers from two herds (MH, n=239; SS, n=330) were wintered at a single location. After a prebreeding examination revealed that 55 heifers had a reproductive tract score (RTS) of 1 (infantile reproductive tracts), they were culled and the remaining heifers were assigned randomly to one of three treatment groups: administration of 25mg PGF2alpha i.m. on Days -12 and 0 followed by estrus detection and insemination between 10 and 14 h after an observed estrus (Control; n=173); administration of 100 microg GnRH i.m. on Day -6, followed by 25 mg PGF2alpha i.m. on Day 0, then fixed-time AI and administration of 100 microg GnRH i.m. on Day +2 (GPG; n=172); and, treatment as for group GPG in addition to administration of 100 microg GnRH i.m. on Day -12 (GGPG; n=169). Bulls were introduced 10 days after AI for 60 days to breed heifers which did not conceive after AI (clean-up bulls). On Days -12, 6, and 0 transrectal ultrasonography was used to monitor ovarian structures in a subset of heifers (30 per treatment). At 30-35 days after AI, ultrasound was used to determine the presence of a viable fetus. Presence of a fetus and stage of pregnancy were determined via palpation per rectum 61-63 days after the conclusion of the breeding season. Heifers in the MH herd (309+/-1.9 kg) were heavier (P<0.001) than those in the SS herd (283+/-1.7 kg) at initiation of the breeding season. Synchronized pregnancy rates were greater (P<0.05) in GGPG (25.4%) and GPG (22.1%) than Control (12.7%) heifers. Pregnancy rates were 9, 21, 32, or 31% for heifers with RTS of 2, 3, 4, or 5, respectively. The average diameter of 22 follicles induced to ovulate in heifers treated with GnRH (GPG and GGPG treatments) was 14.2+/-0.8 mm (range=10.0-23.6 mm). In conclusion, a fixed time ovulation synchronization program using GnRH and PGF2alpha improved pregnancy rates in peripuberal, lightweight replacement beef heifers. PMID- 12566156 TI - Cryopreservation of goat oocytes and in vivo derived 2- to 4-cell embryos using the cryoloop (CLV) and solid-surface vitrification (SSV) methods. AB - This study evaluated the efficiency and toxicity of two cryopreservation methods, solid-surface vitrification (SSV) and cryoloop vitrification (CLV), on in vitro matured oocytes and in vivo derived early stage goat embryos. In the SSV method, oocytes were vitrified in a solution of 35% ethylene glycol (EG), 5% polyvinyl pyrrolidone (PVP), and 0.4% trehalose. Microdrops containing the oocytes were cryopreserved by dropping them on a cold metal surface that was partially immersed in liquid nitrogen. In the cryoloop method, oocytes were transferred onto a film of the CLV solution (20% DMSO, 20% EG, 10mg/ml Ficoll and 0.65 M sucrose) suspended in the cryoloop. The cryoloop was then plunged into the liquid nitrogen. In vivo derived embryos were vitrified using the same procedures. The SSV microdrops were warmed in a solution of 0.3M trehalose and those vitrified with CLV were warmed with incubation in 0.25 and 0.125 M sucrose. Oocytes and embryos vitrified by the SSV method had a significantly lower survival rate than the control (60 and 39% versus 100%, respectively; P<0.05), while the survival rate of CLV oocytes and embryos (89 and 88%, respectively) did not differ from controls. Cleavage and blastocyst rates of the surviving vitrified oocytes (parthenogenetically activated) and embryos (cultured for 9 days) were not significantly different (P>0.05) from the control nor did they differ between vitrification methods. Embryos vitrified with the CLV method gave rise to blastocysts (2/15). Our data demonstrated that the two vitrification methods employed resulted in acceptable levels of survival and cleavage of goat oocytes and embryos. PMID- 12566157 TI - Vitrification of rat embryos at various developmental stages. AB - The effect of developmental stage on the survival of cryopreserved rat embryos was examined. Wistar rat embryos at various developmental stages were vitrified by a 1-step method with EFS40, an ethylene glycol-based solution, or by a 2-step method with EFS20 and EFS40. After warming, the survival of the embryos was assessed by their morphology, their ability to develop to blastocysts (or expanded blastocysts for blastocysts) in culture, or their ability to develop to term after transfer. Most (91-100%) of the embryos recovered after vitrification were morphologically normal in all developmental stages. However, the developmental ability of 1-cell embryos was quite low; exposing them to EFS40 for just 0.5 min decreased the in vitro survival rate from 76 to 9%. The survival rates of 2-cell embryos and blastocysts, both in vitro and in vivo, were significantly higher with a 2-step vitrification process than with a 1-step vitrification process. Very high in vitro survival rates (94-100%) were obtained in 4- to 8-cell embryos and morulae in the 1-step method. Although survival rates in vivo of 4-cell (40%) and 8-cell (4%) embryos vitrified by the 1-step method were comparatively low, the values were similar to those obtained in non vitrified fresh embryos. When morulae vitrified by the 1-step method were transferred to recipients, the in vivo survival rate (61%) was high, and not significantly different from that of fresh embryos (70%). These results show that rat embryos at the 2-cell to blastocyst stages can be vitrified with EFS40, and that the morula stage is the most feasible stage for embryo cryopreservation in this species. PMID- 12566158 TI - Semen characteristics of genetically identical quadruplet bulls. AB - Modern cloning methods have become an important technology in artificial insemination which is used to create and maintain pools of genetically superior bull semen. In this study, semen from four identical quadruplet bulls (Q(1), Q(2), Q(3), and Q(4)) produced by blastomere separation was analyzed to evaluate the differences in reproductive potential, if any, that existed between the identical quadruplet siblings. Analysis of fresh semen collected from 1994 to 1996, showed lower progressive motility and lower sperm concentration for one bull (Q(3)) compared to his identical brothers (P<0.05). Semen characteristics following freezing-thawing procedures have also been tested for these quadruplet bulls. The percentage of motility, progressive motility, and mean path velocity were lower in Q(4) compared with Q(1). Moreover, intracellular calcium level and P25b level (P25b is a sperm surface protein proposed to be a potential bull fertility marker) were lower in Q(4) compared with his siblings (P<0.05). Cryodamage to Q(4)'s frozen-thawed spermatozoa were confirmed by a lower percentage of embryo development after in vitro fertilization. Thus, the higher instability of cryopreserved spermatozoa from Q(4) and the lower semen production of Q(3), compared to their siblings, indicate that differences in semen characteristics can indeed exist among genetically identical animals produced by blastomere separation. PMID- 12566159 TI - Enucleolation of porcine oocytes. AB - Germinal vesicles (GVs) in immature mammalian oocytes contain prominent nucleoli whose role in the process of oocyte maturation is not fully understood. Here we report that the microsurgical removal of nucleoli from immature fully grown porcine oocytes permits germinal vesicle breakdown and chromosome condensation and the enucleolated oocytes mature up to the second metaphase. Interestingly, the enucleolation of growing oocytes which, although unable to mature, resulted in germinal vesicle breakdown and the formation of a cluster of condensed chromatin. These results indicate that the nucleolus in fully grown oocytes is dispensable at least for nuclear maturation. On the other hand, the results obtained in growing oocytes suggest the role of the nucleolus in the cell cycle regulation. PMID- 12566160 TI - Size of the human corpus callosum is genetically determined: an MRI study in mono and dizygotic twins. AB - The factors determining the large variation seen in human corpus callosum (CC) morphology are as yet unknown. In this study heritability of CC size was assessed by comparing the concordance of CC midsagittal area in 14 monozygotic and 12 dizygotic twin pairs with a mean age of 27 years, using magnetic resonance imaging and various methods of calculating trait heritability. Heritability was high regardless of method of assessment. The application of a structural equation model resulted in the estimate that 94% of the variance in CC midsagittal size is attributable to the genome. This indicates that under normal conditions and before the effects of normal aging, there is very modest influence of the environment on CC morphology. The results suggest that correlates of CC size, such as the pattern of cerebral lateralization, cognitive abilities and neuropsychiatric dysfunction may be associated with the genetic determinants of CC morphology. PMID- 12566161 TI - Expression of mBRI2 in mice. AB - Mutations in the BRI(2) gene cause the autosomal dominant neurodegenerative diseases familial British dementia (FBD) and familial Danish dementia (FDD). BRI(2) is a member of a family of type 2 integral transmembrane spanning proteins, including mBRI(2), its murine homologue. The function of BRI(2) is unknown. Northern and Western analyses and in situ hybridization were employed to determine the expression of mBRI(2) in the mouse. mBRI(2) mRNA was expressed in several tissues including the liver, heart, lung, and ubiquitously throughout the brain. mBRI(2) protein was detected at high levels in many brain regions. Murine BRI(2) expression is similar to that described in the human brain but does not fully explain the distribution of pathology seen in FBD and FDD. PMID- 12566162 TI - Flurbiprofen inhibits capsaicin induced calcitonin gene related peptide release from rat spinal cord via an endocannabinoid dependent mechanism. AB - Calcitonin gene related peptide (CGRP) is involved in nociceptive transmission and modulation at the spinal level. In the spinal superperfusion model, Delta(9) tetrahydrocannabinol inhibited capsaicin induced CGRP release in a concentration dependent manner. Similarly, flurbiprofen (3 microM) inhibited spinal CGRP release. This inhibition was reversed by the CB(1) antagonist AM-251 (1 microM), but not by co-administration of prostaglandin E(2) (PGE(2); 285 nM). AM-251 had no modulatory effect on flurbiprofen-induced cyclooxygenase (COX) inhibiting capacity as shown by PGE(2) levels. Furthermore, the phospholipase A(2) inhibitor palmityl trifluromethyl ketone (15 microM) reversed flurbiprofen's inhibitory effect. In conclusion the present work provides evidence on the shift of arachidonic acid metabolism towards endocannabinoids formation in response to COX inhibition as a mechanism for flurbiprofen inhibitory effect on spinal CGRP release. PMID- 12566163 TI - Up-regulation of mu-opioid receptor-mediated G-protein activation in protein kinase Cgamma knockout mice following repeated naloxone treatment. AB - The aim of the present study was to investigate whether repeated treatment with the mu-opioid receptor antagonist naloxone could affect G-protein activation induced by a selective mu-opioid receptor agonist [D-Ala(2),N-MePhe(4),Gly ol(5)]enkephalin (DAMGO) in mice lacking the protein kinase Cgamma isoform monitoring guanosine-5'-o-(3-[(35)S]thio)triphosphate ([(35)S]GTPgammaS) binding. Repeated s.c. administration of naloxone for 7 days resulted in a significant enhancement of the increased [(35)S]GTPgammaS binding by DAMGO to membranes of the spinal cord obtained from mice lacking the protein kinase Cgamma isoform. Furthermore, immunoreactivities of membrane-located protein kinase Cgamma and phosphorylated-protein kinase C in the spinal cord of ICR mice were not altered by repeated naloxone treatment. The present data provide direct evidence that protein kinase Cgamma is not involved in the development of the up-regulation of mu-opioid receptor functions to activate G-proteins in the mouse spinal cord by repeated naloxone treatment. PMID- 12566164 TI - Topiramate protects against motor neuron degeneration in organotypic spinal cord cultures but not in G93A SOD1 transgenic mice. AB - Topiramate is a novel anti-convulsant, structurally distinct from other known anti-convulsants. A number of independent studies suggest that topiramate has anti-excitotoxic properties. It has been found to diminish release of glutamate from neurons and block (-amino-3-hydoxy-5-methylisoxazole-4-proprionic acid glutamate receptor evoked currents. Since activation of non-N-methyl-D-aspartate glutamate receptors is thought to play a role in the selective loss of motor neurons in amyotrophic lateral sclerosis (ALS), we determined whether topiramate could protect against chronic glutamate-mediated motor neuron degeneration. An organotypic spinal cord culture system was used in which glutamate transport is inhibited by pharmacological blockade. After 3 weeks of treatment, topiramate was found to significantly prevent motor neuron degeneration in this culture model. However, the drug did not increase survival in G93A SOD1 transgenic mice, an animal model of ALS. These studies suggest that topiramate could be useful as a neuroprotectant, but were not effective in more complex motor injury paradigms such as the mouse model of ALS. PMID- 12566165 TI - Hypothalamic co-localization of substance P receptor and transneuronal tracer from the lordosis-relevant lumbar epaxial muscles in the female rat. AB - Previous studies indicated that within the ventromedial hypothalamus (VMH), the estrogen receptor alpha (ERalpha)-containing neurons express substance P (SP), but do not comprise the majority of projection neurons. The present study tested the hypothesis that projection neurons within the VMH express SP receptors (NK1), allowing responsiveness to signals from ERalpha-containing neurons. Pseudorabies virus was transneuronally transported from the lordosis-relevant lumbar epaxial muscles to the VMH, labeling 28% of the NK1-containing neurons in the VMH and surrounding area. Thus, SP may influence sexual behavior through its release from the ERalpha-containing neurons, perhaps synaptically affecting NK1 receptor labeled lordosis-relevant projection neurons within the VMH. PMID- 12566166 TI - Molecular analysis of the oligodendrocyte myelin glycoprotein gene in autistic disorder. AB - We previously observed in four autistic patients a new allele (GXAlu 5) of the GXAlu microsatellite marker located in intron 27b of the neurofibromatosis type 1 (NF1) gene (17q11.2). This large intron contains the OMGP gene, coding for the oligodendrocyte myelin glycoprotein expressed by neurons and oligodendrocytes. In the present work, we analysed the distribution of a coding single nucleotide polymorphism (OMGP62) of the OMGP gene, the nearest gene to the GXAlu marker, in a control population (n=101) and in an autistic group (n=65). We observed no significant difference in allele distribution comparing these two groups (chi(2)=1.81; P=0.179). When distinguishing an autistic group with a developmental quotient (DQ) higher than 30 (n=37) and one with a DQ lower than 30 (n=28), we observed an association between allele A and the group with the highest DQ (P=0.015). We found no other polymorphism using SSCP screening and DNA sequencing in the OMGP coding region in 16 autistic patients bearing OMGP62 allele A. PMID- 12566167 TI - Distinguishing of primary cerebral lymphoma from high-grade glioma with perfusion weighted magnetic resonance imaging. AB - To assess the usefulness of perfusion-weighted echo-planar magnetic resonance imaging in the differential diagnosis of primary supratentorial lymphoma (PCNSL) and glioblastoma (GBM), 12 patients with a PCNSL and 12 with a GBM were examined using a 1.5 T magnetic resonance (MR) imager. With dynamic-susceptibility contrast MR imaging the intensity-time curves of each tumor were analyzed, and we determined the relative regional cerebral blood volume ratios (rrCBV [tumor/contralateral white matter (WM)]) to find out whether these parameters could be used to separate PCNSL from GBM. The maximum rrCBV ratio in the PCNSL was significantly lower than that of the GBM (P<0.0001). Comparing the intensity time curves for the two tumor groups, the PCNSL showed a characteristic type of curve with a significant increase in signal intensity above the baseline due to massive leakage of contrast media into the interstitial space. PCNSL tend to have low maximum CBV ratios and typical intensity-time curves. These two parameters may be useful in distinguishing PCNSL from GBM. PMID- 12566168 TI - Association study of a functional catechol-O-methyltransferase-gene polymorphism and cognitive function in healthy females. AB - Using the Wisconsin Card Sorting Test, it has been determined, that the catechol O-methyltransferase (COMT) Val158Met genetic polymorphism, a functional polymorphism that may affect dopamine metabolism, is associated with prefrontal cognitive function. This study of a cohort of 120 healthy young Chinese females attempted to utilize P300 event-related potentials to replicate this finding and to test the relationship between this COMT polymorphism and cortical physiology. The results demonstrate that subjects bearing the Met/Met homozygote have significantly lower mean P300 latencies than do analogs bearing the Val allele. A significant association between this COMT polymorphism and perseverative errors was not demonstrated in the Wisconsin Card Sorting Test, however. We suggest that, although the COMT Val158Met genetic polymorphism may play a role in cognitive function, ethnicity and testing method may affect the association. Since statistical relationships between P300 components and both the COMT genetic polymorphism and schizophrenic disorders have been demonstrated, it seems reasonable to suggest that this COMT genetic variant may affect the P300 abnormality in schizophrenia. PMID- 12566169 TI - Appearance of prolactin-releasing peptide-producing neurons in the area postrema of adrenalectomized rats. AB - Prolactin-releasing peptide (PrRP) was found to be a novel hypothalamic peptide that stimulates prolactin release in vitro and in vivo. In the normal adult rat brain, PrRP neurons are known to be located in only three areas, i.e. the dorsomedial hypothalamic nucleus, ventrolateral reticular formation; and nucleus of the tractus solitarius in the medulla oblongata. These PrRP neurons project neurites into various brain areas, including regions such as the paraventricular nucleus, supraoptic nucleus, and bed nucleus of the stria terminalis. Both PrRP nerve fibers and a high level of PrRP receptor, UHR-1, mRNA are observed in the area postrema (AP),but no PrRP neurons are detected in the AP of normal rats. In this study, we clearly demonstrated that PrRP-producing cells newly appeared in the AP of adrenalectomized rats by in situ hybridization and immunocytochemistry. Our results suggest that PrRP may have some important roles in the AP of adrenalectomized rats. This is the first report demonstrating the appearance of PrRP-positive cells in the AP. PMID- 12566170 TI - Upregulation of purinergic P2Y receptor-mediated calcium responses in glial cells during experimental detachment of the rabbit retina. AB - To investigate injury-induced alterations of purinergic P2Y receptor-mediated calcium responses in glial (Muller) cells of the rabbit retina, neural retinae were experimentally detached from the pigment epithelium. The ATP-evoked calcium responses were recorded in the endfeet of glial cells at the vitread surface of retinal wholemounts. In control retinae, approximately 7% of the glial cells investigated showed ATP-evoked calcium responses. Within 24 h of detachment, significantly more retinal glial cells (42%) showed calcium responses, and glial ATP responsiveness increased further in retinae which were detached for 48 (44%) or for 72 h (64%). The results indicate that in the detached retina, glial cells upregulate their responsiveness to extracellular ATP within 24 h of injury. Thus, P2Y receptor-mediated signalling may be involved in the early steps of glial response to retinal injury. PMID- 12566171 TI - Soya phytoestrogens change cortical and hippocampal expression of BDNF mRNA in male rats. AB - Adult male hooded Lister rats were either fed a diet containing 150 microg/g soya phytoestrogens or a soya-free diet for 18 days. This concentration of phytoestrogens should have been sufficient to occupy the oestrogen-beta, but not the oestrogen-alpha, receptors. Using in situ hybridisation, significant reductions were found in brain-derived neurotrophic factor (BDNF) mRNA expression in the CA3 and CA4 region of the hippocampus and in the cerebral cortex in the rats fed the diet containing phytoestrogens, compared with those on the soya-free diet. No changes in glutamic acid decarboxylase-67 or glial fibrillary acidic protein mRNA were found. This suggests a role for oestrogen-beta receptors in regulating BDNF mRNA expression. PMID- 12566172 TI - Effect of serotonin depletion on the neuronal, endocrine and behavioural responses to corticotropin-releasing factor in the rat. AB - Interactions between serotonin and corticotropin-releasing factor (CRF) have been demonstrated by various studies in different parts of the brain. Both are activated by stressful stimuli. Additionally, serotoninergic fibres directly synapse with the parvocellular division of the paraventricular nucleus (PVN) that mainly synthesize CRF. The functional impact of this serotonin-CRF interaction on CRF-induced responses remains unclear. CRF infusion into the brain evokes a specific pattern of behavioural, endocrine and neuronal changes that resemble those following various forms of stress. The aim of the present study was to investigate the effects of serotonin depletion on acute CRF-induced c-fos expression, corticosterone levels and behavioural responses. Lateral ventricular (i.c.v.) infusion of CRF (250 pmol) resulted in a significant increase in grooming, corticosterone and c-fos mRNA in the PVN. Adequate and specific depletion of serotonin using 5,7-DHT did not alter these CRF-induced changes. These data suggest that acute responses induced by i.c.v. CRF are independent of basal levels of serotonin. PMID- 12566173 TI - A comparison between the distinct inward currents activated in rat cultured dorsal root ganglion neurones by intracellular flash photolysis of two forms of caged cyclic guanosine monophosphate. AB - Whole cell inward currents activated by intracellular photorelease of cyclic guanosine monophosphate (cGMP) were investigated in cultured dorsal root ganglion (DRG) neurones. The actions of two distinct types of caged cGMP (NPE-caged cGMP and a highly water-soluble caged cGMP) were compared. Rapidly activating inward currents were evoked by cGMP in a subpopulation (12.5%) of neurones and these currents may be due to activity of cyclic nucleotide-gated channels. In contrast in 52% of DRG neurones intracellular photorelease of cGMP activated a delayed Ca(2+)-dependent inward current through the generation of cyclic ADPribose and mobilisation of Ca(2+) from ryanodine sensitive intracellular stores. Similar delayed inward currents were activated by both caged compounds but only NPE-caged cGMP evoked rapidly activating currents. Cyclic GMP appears to increase excitability in some DRG neurones by diverse mechanisms. PMID- 12566174 TI - Anisomycin inhibits the late maintenance of long-term depression in rat hippocampal slices in vitro. AB - Studies were performed to investigate whether electrically-induced long-term depression (LTD) within rat hippocampal slices in vitro shares any common cellular features with LTD in the intact animal, with particular emphasis being placed on mechanisms required for its late maintenance. Our initial studies have led to the development of stimulation protocols which are able to reliably produce different forms of LTD. Depending on the induction protocol applied, we are able to demonstrate a transient protein synthesis-independent early-LTD with a duration of up to 3-4 h, together with a de novo protein synthesis-dependent late-LTD lasting for at least 8 h. Furthermore, we are able to show input specific LTD within the CA1 region, with expression shown only by those synapses specifically stimulated by a low-frequency protocol. These studies are important pre-requisites to investigate mechanisms of 'synaptic tagging' and 'late associativity' during LTD. PMID- 12566176 TI - PAC1 receptors in chick cerebral cortex: characterization by binding of pituitary adenylate cyclase-activating polypeptide, [125I]-PACAP27. AB - In this study we characterized receptors for pituitary adenylate cyclase activating polypeptide (PACAP) in chick cerebral cortex by in vitro binding technique, using [125I]-PACAP27 as a ligand. The specific binding of [125I] PACAP27 to chick cerebral cortical membranes was found to be rapid, stable, saturable, and of high affinity. Scatchard analysis suggested binding to a single class of receptor binding sites with high affinity (K(d)=0.41+/-0.08 nM) and high capacity (B(max)=457+/-35 fmol/mg protein). The relative rank order of potency of the tested peptides to inhibit [125I]-PACAP27 binding to chick cerebrum was: PACAP38 approximately PACAP27>PACAP6-27 approximately PACAP6-38 >> chicken VIP >> mammalian VIP >> secretin (inactive). It is concluded that the cerebral cortex of chick, in addition to VPAC recognition sites, contains a large population of PAC(1)-type receptor binding sites. PMID- 12566175 TI - A randomised, double blind, placebo controlled crossover study of the cholecystokinin 2 antagonist L-365,260 as an adjunct to strong opioids in chronic human neuropathic pain. AB - The aim of this study was to establish if the cholecystokinin (CCK) 2 antagonist L-365,260 augments the analgesic effect of morphine in human subjects with chronic neuropathic pain. This is a randomised, double blind, placebo controlled study of 40 adult subjects taking morphine for neuropathic pain. Each received placebo, L-365,260 30 mg and L-365,260 120 mg in three divided doses daily separated by a washout period in random order. Pain, activity, sedation, sleep and side effects were recorded along with 12 lead ECGs, renal and liver function tests and full blood pictures. L-365,260 failed to augment the analgesic effect of morphine at any of the dose levels used. Side effects were minor. There were no changes in ECGs and biochemical indices were unaltered with its use. The CCK 2 antagonist L-365,260 does not augment the analgesic effect of morphine in subjects with chronic neuropathic pain. L-365,260 was well tolerated and side effects from its use were minor. PMID- 12566177 TI - Cerebral glucose metabolism, cerebrospinal fluid-beta-amyloid1-42 (CSF-Abeta42), tau and apolipoprotein E genotype in long-term rivastigmine and tacrine treated Alzheimer disease (AD) patients. AB - We evaluated cerebral glucose metabolism (CMRglc) and cerebrospinal fluid (CSF) levels of tau and beta-amyloid(1-42) (Abeta42), in relation to apolipoprotein E (ApoE) genotype, in patients with mild Alzheimer disease (AD) treated with rivastigmine (n=11) and tacrine (n=16) for 1 year; and two untreated AD groups. The rivastigmine-treated AD patients showed a significant increase in CMRglc as compared to both tacrine-treated and untreated AD subjects. The rivastigmine treated AD group showed no change in CSF-tau levels after 1 year, while in contrast a significant increase as seen in tacrine-treated and untreated AD patients. The CSF-tau changes were mainly seen in ApoE epsilon4 carriers. There was no significant change in Abeta42 after 1-year treatment with either rivastigmine or tacrine. This study shows that the two long-term cholinesterase inhibitor treatments exert different effects on biological markers for AD. PMID- 12566179 TI - Aged-related differences in the attentional cost of pointing movements. AB - The aim of the present study was to analyze, in children aged 6, 8 and 11, the developmental trend of the attentional cost related to the programming and execution of pointing movements, using a dual-task paradigm. Our results showed that the attentional cost associated to the programming and the first phase of the pointing movement decreased non linearly with age, in particular a plateau between 8 and 11 was observed. This confirmed that the developmental trend of the control of pointing movements is characterized by a no monotonic evolution in which feedforward processes were predominantly used in the younger children, feedback processes were predominantly used in older children (since age 8), and one-line over feedforward processes were used in adults. PMID- 12566178 TI - Neuronal damage by secretory phospholipase A2: modulation by cytosolic phospholipase A2, platelet-activating factor, and cyclooxygenase-2 in neuronal cells in culture. AB - Activation of cytosolic phospholipase A(2) (cPLA(2)) is an early event in brain injury, which leads to the formation and accumulation of bioactive lipids: platelet-activating factor (PAF), free arachidonic acid, and eicosanoids. A cross talk between secretory PLA(2) (sPLA(2)) and cPLA(2) in neural signal transduction has previously been suggested (J Biol Chem 271:32722; 1996). Here we show, using neuronal cell cultures, an up-regulation of cPLA(2) expression and an inhibition by the selective cPLA(2) inhibitor AACOCF3 after exposure to neurotoxic concentrations of sPLA(2)-OS2. Pretreatment of neuronal cultures with recombinant PAF acetylhydrolase (rPAF-AH) or the presynaptic PAF receptor antagonist, BN52021, partially blocked neuronal cell death induced by sPLA(2)-OS2. Furthermore, selective COX-2 inhibitors ameliorated sPLA(2)-OS2-induced neurotoxicity. We conclude that sPLA(2)-OS2 activates a neuronal signaling cascade that includes activation of cPLA(2), arachidonic acid release, PAF production, and induction of COX-2. PMID- 12566180 TI - Electron microscopic analysis of nanoparticles delivering thioflavin-T after intrahippocampal injection in mouse: implications for targeting beta-amyloid in Alzheimer's disease. AB - Prevention of beta-amyloid (Abeta) production, aggregation and formation of Abeta deposits is a key pharmacological target in Alzheimer's disease. The passage of Abeta-binding compounds through the blood-brain barrier is often hampered for free ligands, whereas it is enhanced by drug encapsulation in nanoparticles. Here, we describe the preparation and characterization of polymeric carriers containing thioflavin-T as a marker for fibrillar Abeta. This study is then focused on electron microscopic analyses of thioflavin-T after injection of thioflavin-T-containing nanoparticles into the mouse hippocampus. Therefore, the photoconversion of fluorescent thioflavin-T as model drug was performed in tissues fixed 3 days post-injection. Thioflavin-T delivered from nanospheres was predominantly found in neurons and microglia. Our data suggest that drugs delivered by nanoparticles might target Abeta in the brain. PMID- 12566181 TI - Autotransfusion for ruptured ectopic pregnancy. AB - OBJECTIVES: Clinical outcomes following the exclusive use of autotransfusion in the management of ruptured ectopic pregnancy are reviewed. METHODS: A MEDLINE search (1966-2002) for relevant articles documenting the exclusive use of autotransfusion, and data collection and analysis was made. RESULTS: There were 21 studies, 16 from developing and five from developed countries, involving 632 cases of ruptured ectopic pregnancies. Hypovolemic shock with significant hemoperitoneum (>500 ml) was the most common complication. The procedure was performed completely manually in developing countries and with the help of a device in developed countries. The mean volume of autotransfused blood was over 1000 ml, with mean hemoglobin levels ranging from 6 to 12.5 g/dl. Mean posttransfusion hemoglobin levels were higher than pretransfusion levels. There was one death, thought to be due to pulmonary embolism, and nine major and minor complications. CONCLUSIONS: Autotransfusion is useful in the management of ruptured ectopic pregnancy. PMID- 12566182 TI - Calcium-phosphorus-magnesium homeostasis in pregnant women after renal transplantation. AB - OBJECTIVE: The aim of the study was the assessment of calcium-phosphorus magnesium homeostasis in pregnant women after renal transplantation. METHODS: The study covered 64 pregnant women in the third trimester of gestation including: 33 women after renal transplantation (the study group) and 31 healthy pregnant women (the control group). Women from both groups were at the similar age: 30.8+/-4.7 vs. 31.3+/-5.0 years (NS) and at the same gestational age 34.8+/-2.4 vs. 35.3+/ 2.6 weeks (NS). The mean body mass index (BMI) in the women from the study group before pregnancy was 21.49+/-2.81 vs. 22.1+/-3.02 in the control group (NS), BMI before delivery was 25.43+/-3.05 vs. 26.0+/-3.35 (NS), the percentage of the BMI increase during pregnancy was 18.7+/-7.68 vs. 17.65+/-7.13 (NS) and BMI increase during gestation was 3.93+/-1.56 vs. 3.90+/-1.54, respectively (NS). Arterial blood pressure at the time of blood samples collection for biochemical tests was 151.4+/-26.8/92.5+/-16.9 in women from the study group comparing to 115.0+/ 6.0/68.0+/-7.0 mmHg (P<0.001) in the patients from the control group. The maximal blood pressure during pregnancy was 169.2+/-20.7/102.7+/-14.0 vs. 118.0+/ 7.0/70.0+/-8.0 mmHg (P<0.001), respectively. We estimated serum levels of: total Ca, ionized Ca(2+), inorganic phosphorus (P(i)), Mg, total protein, albumin and blood morphology. Moreover, urine levels of Ca, P(i), Mg and protein were assessed. RESULTS: The pregnant women after renal transplantation presented increases in serum concentrations of total Ca (2.54+/-0.20 vs. 2.16+/-0.10 mmol/l; P<0.001) and ionized Ca(2+) (1.322+/-0.104 vs. 1.12+/-0.07 mmol/l; P<0.001) and the decrease in P(i) level (1.013+/-0.211 vs. 1.10+/-0.16 mmol/l; P<0.05), total protein (59.3+/-7.0 vs. 65+/-5 g/l; P<0.001) and albumin (461.6+/ 65.65 vs. 493.2+/-59 micromol/l; P<0.05). Moreover, in the study group drop in red blood cells count to 3.71+/-0.56 vs. 4.01+/-0.35 x 10(12)/l (P<0.02) in the control group was detected. Despite increased volume of 24-h urine collection in the kidney recipients we observed significantly decreased urine 24-h calcium excretion 2.47+/-0.92 vs. 6.72+/-3.49 mmol (P<0.001) and simultaneous increase in urine Mg excretion 3.422+/-1.025 vs. 2.18+/-0.52 mmol/24 h (P<0.001). There was no difference in urine 24-h P(i) excretion between the study and the control group. The pregnant renal transplant recipients presented proteinuria of 1.19+/ 1.9 g/24 h. CONCLUSIONS: Women after kidney grafting present vital aberrations in calcium-phosphorus-magnesium homeostasis during pregnancy. The most significant changes are associated with calcium metabolism (high increase in serum Ca levels and impairment of renal elimination of calcium). The observed changes may be influenced by the doses of immunosuppressive agents and disturbed renal function. PMID- 12566183 TI - Ambulatory epidural anesthesia and the duration of labor. AB - OBJECTIVES: Ambulatory epidural analgesia has become a common option for women in labor in France. We tested the hypothesis that a method of epidural analgesia that allowed women to walk had specific advantages regarding mode of delivery, consumption of local anesthetic, oxytocin requirement, and labor duration. METHODS: Two hundred and twenty-one women with uncomplicated pregnancies who presented in spontaneous labor between 36 and 42 weeks of gestation or who were scheduled for induced labor were randomly divided into two groups, ambulatory and non-ambulatory. All were given intermittent epidural injections of 0.1% ropivacaine with 0.6 microg/ml sufentanil for analgesia during labor (P<0.05 was considered significant). None of the women had previous cesarean delivery. RESULTS: There were no significant differences between the two groups in mode of delivery, consumption of local anesthetic, or oxytocin requirement. However, a significant difference was noted in labor duration (173.4+/-109.9 min vs. 236.4+/ 130.6 min; P=0.001). CONCLUSIONS: Walking with ambulatory labor analgesia shortens labor duration but has no other effect on the progress and outcome of labor. PMID- 12566184 TI - The prevalence and timing of cervical cerclage placement in multiple gestations. AB - OBJECTIVES: To investigate the prevalence and timing of cervical cerclage placement in multiple gestations. METHODS: Our perinatal database was queried for all multiple gestations delivered at Evanston Hospital from 12/95 through 12/00. This list was then cross-matched with billing and medical records for 'incompetent cervix' and 'cerclage.' The medical records of all deliveries /=14 weeks over a 5-year period. The number of patients that underwent cerclage placement was 29 or 3.6%. The mean gestational age at cerclage placement was 18.6+/-4.5 weeks (range 11-24.6). Twelve were elective or prophylactic while 17 were 'urgent' or 'emergent.' The mean gestational age for the 17 emergent cerclages was 21.4+/-2.2 weeks (range 16.6-24.6). When compared with those patients who did not undergo cerclage placement, there was no difference in maternal demographics including age, parity, or previous full-term delivery. There was a significant difference in the gestational age at delivery for the cerclage vs. no cerclage group; 29.3+/-5.6 vs. 34.4+/-4.6 weeks, respectively, and in the frequency of losses at 0.05). Two of the seven affected patients did not have maternal serum biochemical measurements due to the immediate termination of pregnancies. CONCLUSIONS: We demonstrated that pregnancies with fetal acrania could be easily identified at the time of NT scanning. Careful ultrasound inspection of fetal structure during NT measurements at 10-13 weeks of gestation provides an encouraging advantage for early diagnosis of fetal acrania. PMID- 12566187 TI - Parametrial tumor spreading patterns in cervix cancer patients treated by radical hysterectomy. AB - OBJECTIVES: To evaluate tumor-spreading patterns in the parametrium. METHODS: We conducted a prospective clinical trial between January 1998 and December 2000 to define a new method for parametrium evaluation. The parametrium was divided into three areas, paracorpus, paracervix, and paravagina. A total of 284 consecutive patients with FIGO stage IB to IIA cervical cancer who had undergone radical hysterectomy were considered for the study. RESULTS: Of the 262 patients who were found eligible for evaluation, 135 had histopathologic analysis performed according to the new method and 127 with the traditional method. The detection of rate of parametrial invasion was 36 (26.7%) with the new and 13 (10.2%) with the traditional method (P=0.0014). The frequency of pelvic lymph node metastasis was 66.7% in patients who had tested positive for invasion of the paracorpus, 57.7% in those who had tested positive for invasion of the paracervix, and 71.4% in those who had tested positive for invasion of the paravagina. The frequency of pelvic lymph node metastasis in patients who had tested negative for invasion of the paracorpus, paracervix, or paravagina was 4.0%. Tumor cells tend to spread laterally and inferiorly in the parametrium. CONCLUSIONS: Using our classification of three parametrium areas for histologic examination can increase the detection rates of parametrial tumor invasion and help prevent failure of local treatment by allowing to implement appropriate adjuvant therapy. PMID- 12566189 TI - Postcoital contraception in Turkey. AB - OBJECTIVES: The purpose of this article is to determine the knowledge status of women concerning postcoital contraception when they turn to curettage in order to terminate an undesired pregnancy. METHODS: This defining study has been carried out at a maternity hospital in Izmir on pregnant women admitted for curettage. We interviewed 150 pregnant cases using an improbability sampling method. RESULTS: We discovered that 48.7% of women had avoided pregnancy before curettage by withdrawal before ejaculation. When we investigated the method used after uncontrolled coitus, 27.3% of cases reported vaginal douche while 12.7% indicated curettage. After having an unprotected coitus or failure in contraceptive methods 99.3% of the women indicated they would report to health institutes to utilize postcoital methods if they were familiar with them. CONCLUSIONS: Postcoital contraception may be a solution for undesired pregnancies. PMID- 12566188 TI - Comparison of fennel and mefenamic acid for the treatment of primary dysmenorrhea. AB - OBJECTIVES: To compare the effect of Foeniculum vulgare variety dulce (Sweet Fennel) vs. mefenamic acid for the treatment of primary dysmenorrhea. METHODS: A cohort of seventy women, 15-24 years old from a local university and high-school, who complained of dysmenorrhea were enrolled in this study. Ten cases were excluded due to evidence of secondary dysmenorrhea. The remaining 60 patients were graded mild, moderate and severe on the basis of a verbal multidimensional scoring system. Thirty patients with mild dysmenorrhea were also excluded from the study. Each of the 30 cases with moderate to severe dysmenorrhea was evaluated for three cycles. In the first cycle no medication was given (control cycle), in the second cycle the cases were treated by mefenamic acid (250 mg q6h orally) and in the third cycle, essence of Fennel's fruit with 2% concentration (25 drops q4h orally), was prescribed at the beginning of the cycle. These cycles were compared day by day for the effect, potency, time of initiation of action and also complications associated with each treatment modality, by using a self scoring system. Intensity of pain was reported by using a 10-point linear analog technique. Statistical analyses were performed by the independent sample t-test, paired t-test and repeated measurement analysis method. RESULTS: In the study group the mean age of menarche was 12.5+/-1.3 years, the mean duration of menstruation was 6.6+/-1.4 days with the mean cycle days of 27+/-3. The findings observed during menses were as follows: headache in 26.7%, nausea in 63.3%, vomiting in 23.3%, diarrhea in 33.3%, fatigue in 93.3% and leaving the daily tasks undone was reported in 86.9% of the cases. Both of the drugs effectively relieved menstrual pain as compared with the control cycles (P<0.001). The mean duration of initiation of action was 67.5+/-46.06 min for mefenamic acid and 75+/ 48.9 min for fennel. The difference was not statistically significant (P=0.57). Mefenamic acid had a more potent effect than fennel on the second and third menstrual days (P<0.05), however, the difference on the other days was not significant. No complication was reported in mefenamic acid treated cycles, but five cases (16.6%) withdrew from the study due to fennel's odor and one case (3.11%) reported a mild increase in the amount of her menstrual flow. CONCLUSIONS: The essence of fennel can be used as a safe and effective herbal drug for primary dysmenorrhea, however, it may have a lower potency than mefenamic acid in the dosages used for this study. PMID- 12566190 TI - Conservative management of the acute abdomen secondary to hemorrhagic disease of the ovary. PMID- 12566191 TI - High bilateral ureteric injury during cesarean birth, and unique repair. PMID- 12566192 TI - Ruptured uterus and the partograph. PMID- 12566193 TI - Pregnancy with a microperforated hymen. PMID- 12566194 TI - A hypothesis for altered activity of insulin-like growth factor I in women with pre-eclampsia. PMID- 12566195 TI - A framework for a new approach to antenatal care. AB - A framework for a new approach to antenatal care (ANC) is presented to improve maternal health. Based on evaluations of ANC, safe motherhood programs, gender and social theory, it suggests that managers should draw upon existing family and community support systems, and develop partnerships beyond the health service. Policy and program changes are required in: professional mandates for ANC providers, organization of ANC services, service protocols, training programs, policy towards TBAs, referral care, and service support systems. PMID- 12566196 TI - Factors influencing the quality of life of infertile women in United Arab Emirates. AB - OBJECTIVES: To measure the quality of life in a representative sample of infertile women and evaluate their sociocultural attitude to this condition. METHODS: Two hundred sixty-nine infertile women attending the Assisted Reproduction clinic, Tawam Hospital were consecutively selected. They were interviewed about the effect of infertility on their quality of life using a structured, measurement-specific and pre-tested questionnaire. RESULTS: Parameters mostly affected were mood-related mainly in women above 30 years, with primary and female factor infertility and those in polygamous marriages. Quality of life did not affect sexual performance and was not affected by duration of infertility or cost of treatment. CONCLUSION: The results highlight the importance of bearing children and the stresses exerted on infertile women in Eastern societies. Thorough counseling and continuing support of infertile women is therefore indicated to improve their quality of life. PMID- 12566197 TI - Attitudes of Singapore women toward cesarean and vaginal deliveries. AB - OBJECTIVES: To assess the attitude, knowledge, and expectations of Asian pregnant women toward cesarean and vaginal deliveries. METHODS: Written questionnaires were given to pregnant women attending the National University Hospital antenatal clinics, and 160 responses were tabulated and analyzed using SPSS software. RESULTS: The participation rate was 65% and 50% of the respondents were Chinese, 20% Indian, 21% Malay, 2% White, and 9.2% Other. The median age was 31 years, and approximately 43% were primiparas. Only 3.7% of them would prefer an elective cesarean delivery, and although 50% had friends or relatives who requested one, only 3% felt that this influenced their preference. The most common reasons for choosing a cesarean delivery were avoiding labor pains and lowering the risk of fetal distress. When asked which modality they would recommend to their friends, only 2% expressed that they would recommend cesarean delivery; however, 71% stated that women should have the right to request a cesarean delivery. Awareness of complications arising from vaginal and cesarean deliveries was generally low and related to the frequency of these complications. There was no significant correlation between demographic characteristics and maternal preference for mode of delivery. CONCLUSIONS: Asian women largely prefer a vaginal delivery, and their attitude toward cesarean delivery on demand is comparable with that of Western women. Cultural or ethnic differences are unlikely to affect maternal preference for cesarean delivery in Singapore women. PMID- 12566198 TI - Editor's comment: The FIGO Save the Mothers Initiative. PMID- 12566199 TI - Saving mothers' lives: the FIGO Save the Mothers Initiative. PMID- 12566200 TI - The FIGO Save the Mothers Initiative: the Uganda-Canada collaboration. AB - In 1998, the ob/gyn associations of Uganda and Canada launched, under the umbrella of the FIGO Save the Mothers Initiative, a district-wide intervention which aimed to increase the availability and utilization of emergency obstetric care (EmOC) services in a rural district of Uganda. The article describes the experience of two professional ob/gyn associations in the development, implementation, monitoring and evaluation of the project. Preliminary results after 24 months of intervention indicate important gains in the capacity of health professionals to deliver EmOC, the availability of emergency transportation services and met need for EmOC. PMID- 12566201 TI - FIGO Save the Mothers Initiative: the Central America and USA collaboration. AB - The American College of Obstetricians and Gynecologists (ACOG) and the Central American Federation of Associations and Societies of Obstetrics and Gynecology (FECASOG), as a part of the FIGO Save the Mothers Initiative, undertook a pilot project to improve provision of basic emergency obstetric care in selected departments in four Central American countries. This article describes the process of the development and implementation of the project. Preliminary results suggest that the capacity to provide this care has been improved by the training of healthcare personnel. PMID- 12566202 TI - Program note: using UN process indicators to assess needs in emergency obstetric services: Morocco, Nicaragua and Sri Lanka. PMID- 12566204 TI - Interdisciplinary preoperative patient education in cardiac surgery. AB - Patient education in cardiac surgery is complicated by the fact that cardiac surgery patients meet a lot of different health care providers. Little is known about education processes in terms of interdisciplinary tuning. In this study, complete series of consecutive preoperative consultations of 51 cardiac surgery patients with different health care providers (physicians, nurses and health educators) were videotaped. The information exchange between patients and providers was analyzed directly from the video recordings by using an adaptation of the Roter Interaction Analysis System (RIAS) and a checklist of relevant informational topics. Results pointed to overlaps and gaps as well as to a lack of a patient-centered approach. The physicians were mostly overlapped by the nurses, who spent almost 30% of the time on talking about medical issues. Gaps were found in giving psycho-educational information and emotional support, needed to establish effective patient education. The findings provided a sound basis for developing guidelines and changes in the organization of the education process. PMID- 12566205 TI - Evaluation of a question-and-answer booklet on early-stage prostate-cancer. AB - A question-and-answer booklet about early-stage prostate-cancer was created for patients and their family members. Two sequential studies were conducted to determine if the single source of information would be useful to both patients and their families. The first study used 1:1 interviews with 11 readers (6 patients and 5 family members) to identify features of the booklet that may be problematic. In all, six features were identified as either irrelevant or problematic in their design. The second study was a survey of 54 patients (79% response rate) and 33 family members (49% response rate); the study was designed to obtain an overall evaluation of the booklet, to clarify the proportion of readers for whom the features identified in the first study were problematic, to provide insight into how and why readers were reading the booklet, and to determine if patients and family differed on any of the outcomes. Results showed that 85% of readers liked the booklet; most (81.6%) read it from beginning to end. Most readers in both groups wanted the information to help them understand (85.2% patients; 87.9% family); more patients than family wanted it for treatment decision making (44.4% patients; 27.3% family) and for planning (35.2% patients; 9.1% family); more family (42.4%) than patients (20.4%) wanted the information to help them provide support. We conclude that even though patients and family differ in why they want information, the booklet appears to be considered useful by both groups. PMID- 12566207 TI - Cognitive-behavioural therapy with a Huntington's gene positive patient. AB - The treatment of choice for depression or anxiety after genetic testing is the use of medication. The present paper reports a case of a lady who had a positive test result for Huntington's disease (HD). After the predictive test her mood declined and she experienced symptoms of anxiety. Cognitive-behavioural therapy (CBT) enabled her to deal effectively with her negative automatic thoughts (NATs) and interpret situations more realistically. Therapy was successful in reducing her level of physical, behavioural and affective symptoms and in increasing her sense of control. These gains were maintained at 3 and 6 months in spite of the death of her mother. The paper discussed the potential value of cognitive behavioural theory and therapy in similar cases. PMID- 12566206 TI - Correlates of women's preferences for treatment of heavy menstrual bleeding. AB - This cross-sectional survey investigated factors associated with treatment preferences of women with menorrhagia. Women (n = 474) aged 35-54 years referred to gynaecology out-patient clinics for menorrhagia were mailed a self administered questionnaire before their first clinic visit. The main outcome measure was treatment preference. Hysterectomy and conservative treatment (combined with no treatment) were favoured equally often. In a multivariate analysis, completed family size (P = 0.003), menstrual pain (P = 0.02), irregular periods (P = 0.03), and higher age (P = 0.04) predicted hysterectomy preference, as did lower education level (P = 0.001), gynaecologist consultations (P = 0.002), and unemployment (P = 0.03). The psychological factors assessed were not associated with treatment preference. In conclusion, rational considerations regarding stage of reproductive life and severity of symptoms were linked to women's treatment preferences. However, education, employment status and use of specialists' services guided women's preferences even more powerfully than menstrual symptoms and burden caused by them. Doctors should pay more attention to motivating women for a trial of conservative treatment since only half of the women reported previous treatment for their menstrual problem. PMID- 12566208 TI - Consumer-oriented evaluation of health education services. AB - The objective of this study was to evaluate the effect of the presence of formal health education units in health plans on the health education reported by the consumer. The research consisted of interviews with health educators in the Israeli health plans and a random sample of telephone interviews with 793 Israeli residents between the ages of 45 and 75. The interviews with consumers included measures of counseling on smoking, physical activity, weight reduction, hypertension and diabetes. Two of the four Israeli health plans have formal health education units. After adjustment for other variables, however, the quality and quantity of counseling reported by the consumers does not differ, in most subjects, between health plans with or without health education units. The presence of a health education unit within a health plan had little effect on the counseling reported by the consumer. The reasons for and implications of the results are discussed. PMID- 12566209 TI - Interaction analysis of physician-patient communication: the influence of trait anxiety on communication and outcome. AB - Little attention has been paid to how patients' personality traits interfere with the communication and the outcome of physician-patient interaction. We performed an experimental study with students with high and low trait anxiety as patients. One physician conducted a single consultation with 41 students applying two beforehand-specified consultation styles. Patients completed questionnaires concerning emotional state and satisfaction. The actual content of the consultations was analyzed by Roter interaction analysis system (RIAS). The physician gave more biomedical information to low-anxiety students than high anxiety students. Students who provided a lot of biomedical information themselves were less tense after the consultation. However, students with high anxiety were more dependent on the physician actively asking biomedical questions for them to be able to deliver that same information. In contrast to low-anxiety students, those with high anxiety were less satisfied after consultations involving many psychosocial questions posed by the physician and a good deal of emotional talk on their own part. Compared to low-anxiety students, students with high anxiety were less satisfied and tenser after consultations with much positive emotional talk on the part of the physician. We conclude that physicians and educators should be aware that psychological and emotional communication may be experienced as intrusive and inappropriate by patients with high trait anxiety when they present minor somatic problems. PMID- 12566210 TI - Participant reactions to a computerized telephone system for nutrition and exercise counseling. AB - This paper reports on an evaluation of the reactions of participants in a computer-controlled telephone conversation system (telephone-linked care, TLC) designed to offer nutrition and exercise counseling. After 6 months in the study, subjects were asked a series of questions about their opinions of the TLC system, including overall satisfaction and the system's helpfulness. One hundred and ninety individuals completed the attitude survey. On a scale of 0-100, respondents rated the overall satisfaction and helpfulness of the system at 63.6 and 62.3. Subjects using the nutrition counseling version of TLC rated it significantly higher on satisfaction (73.0 versus 52.4) and helpfulness (70.3 versus 53.7) than did subjects using the exercise version. Satisfaction and helpfulness were correlated with perceived usability, amount of contact, realism, and credibility (P < 0.01). Multivariate analyses showed that treatment group and number of calls made accounted for the greatest amount of variance in ratings of satisfaction and helpfulness. The findings suggest that the amount of contact with this technology, reflected by the number of calls, and the treatment group, nutrition or exercise, were significant predictors of reported satisfaction and perceived helpfulness of the system. PMID- 12566211 TI - Reproductive health perceptions, beliefs and sexual risk-taking among youth in Addis Ababa, Ethiopia. AB - Appropriate knowledge and beliefs about sexuality and necessary life skills do not often precede sexual behaviours among young people. This study outlines a profile of sexual risk factors and the underlying sexual and reproductive health perception and beliefs among youth (aged 15-24 years) in Addis Ababa, Ethiopia. Five hundred and sixty-one youth residing in two of the six zones of Addis Ababa filled a self-administered questionnaire. Three outcome variables: history of sexual activity in the past, use of condom during recent sexual intercourse and use of alcohol, were used to construct a pyramid of sexual risk categories. Being a female, age group of 20-24 years and out-of-school significantly increased the likelihood of belonging to the most vulnerable category. Although young people were heterogeneous in their sexual risk-taking, they were equally uninformed on reproductive health matters related to puberty and sexuality. Their respective beliefs also tended to be flexible and unstable to consistently predict their behaviour. Possible reasons for such heterogeneity and its practical implications in sex education strategies are discussed. PMID- 12566212 TI - Interactive group education for refugees from the Former Yugoslavia to reduce their oil consumption. AB - In 1999 at an outpatient dietetic consultation in Geneva, Switzerland, 20% of the patients were refugees. Most were from the Former Yugoslavia. The main goal of dietary counseling was to decrease their fat consumption. As one-on-one counseling was unsuccessful, a pilot intervention was developed to explore the potential of participatory workshops for identifying and implementing appropriate behaviors. Four series of three workshops were held. Participants identified and practiced ways to reduce fat consumption. Knowledge skills were measured on completion of the workshops and 6 months later. Oil was highly valued in participants' representation of health. However, average reduction of oil per recipe was 58%, or 35 ml (95% CI: 15-55). The application of oil-sparing techniques increased nine-fold. Such an approach may be an alternative to one-on one dietary counseling where this is ineffective. Next steps should include a trial to determine the impact of new skills on actual fat consumption. PMID- 12566213 TI - Inequalities in the provision and monitoring of primary care counselling: views from Wiltshire, England. AB - This study reviewed the provision of counselling services through 30 general practices in Wiltshire, a rural county in South West England, by means of a postal questionnaire, which had an 80% response rate, and interviews with key stakeholders. While 83% of general practices had a practice-based counsellor and 87% referred patients to counselling agencies, significant inequities were identified across the area for the level of counselling provision. Systems for the monitoring, management and evaluation of counselling services were found to be inadequate and concerns were raised regarding the absence of standardised practice guidelines. These findings lend support to the Government's intention to pursue statutory regulation of psychotherapists, counsellors and related groups working in the NHS. PMID- 12566214 TI - Knowledge and attitudes of gynecologists regarding genetic counseling for hereditary breast and ovarian cancer. AB - In a survey we investigated whether gynecologists are sufficiently knowledgeable to perform genetic counseling. It provides information for the development and evaluation of a counseling manual for professionals in primary health care. The members of the sample, consisting of 529 gynecologists in northern Germany, were mailed a questionnaire concerning their knowledge of and attitudes towards genetic counseling and testing for hereditary breast and ovarian cancer (HBOC). The response rate was 32.5% (n = 172). The majority of the respondents (82%) have received requests from patients for genetic testing. Most would offer basic genetic counseling to their patients, 66% feel knowledgeable enough to do so. Physicians set high value on communicating clinical management options, but also consider psychosocial aspects to be important. The results suggest that HBOC genetics play a noticeable role in the practice of gynecology in Germany. There is consensus about the need for further educational training to deal with cancer genetics in physicians' daily practice. PMID- 12566215 TI - Counselling interventions to address the psychological consequences of screening mammography: a randomised trial. AB - We examined the effectiveness of offering counselling to women undergoing screening mammography who are recalled for further investigations that do not lead to a diagnosis of cancer. Women were randomised to being offered either face to-face (n = 66) or telephone counselling (n = 68) or usual care (n = 71) at the recall clinic after being told that their screen-detected abnormality was not cancer. The PCQ (a reliable and valid measure of the psychological consequences of screening mammography) measured the emotional, social and physical functioning of women at the recall clinic (Time 1) and after the counselling intervention (Time 2). Analyses of covariance (ANCOVA) showed no main effects for intervention on Time 2 levels of functioning after adjustment for the respective covariate of Time 1 functioning. Time 1 levels significantly predicted Time 2 levels of functioning. When data were analysed according to whether women actually received any type of counselling versus not receiving counselling, participation in counselling was associated with lower scores on dysfunction scales at Time 2, after adjusting for Time 1 levels. PMID- 12566216 TI - Pharmacological modulation and differential regulation of the cardiac gap junction proteins connexin 43 and connexin 40. AB - Gap junction channels provide the basis for the electrical syncytial properties of the heart as a communicating electrical network. Cardiac gap junction channels are predominantly composed of connexin 40 or connexin 43. The conductance of these channels (g(j)) can be regulated pharmacologically: substances which activate protein kinase C, protein kinase A or protein kinase G may alter Cx43 gap junction conductance. However, for PKC, this seems to be subtype specific. Thus, antiarrhythmic peptides can enhance g(j) via activation of PKCepsilon, while FGF-2 reduces g(j) via PKCepsilon. Lipophilic drugs can uncouple the channels. Besides an acute regulation of g(j), the expression of the cardiac connexins can also be regulated. A decrease in Cx43 with a concomitant increase in Cx40 has been found in end-stage failing hearts, while in renovascular hypertension, an increase in Cx43 has been described. Mediators like endothelin 1, angiotensin-II, TGF-beta, VEGF, and cAMP have been shown to increase Cx43. Interestingly, endothelin-1 and angiotensin-II increased Cx43 but did not affect Cx40 expression. In contrast, in humans suffering from atrial fibrillation (AF), the content in Cx40 can be enhanced while Cx43 was unaltered, although in several other studies, other changes of the cardiac connexins were found, which might be related to the type of AF. Regarding the role of calcium, the content in both Cx40 and Cx43 was decreased in cultured neonatal rat cardiomyocytes after 24 h administration of 100 nM verapamil. Thus, gap junctional channels can be affected pharmacologically either acutely by modulating gap junction conductance or chronically by altering gap junction protein expression. Interestingly, it appears that the expression of Cx43 and Cx40 can be differentially regulated. PMID- 12566217 TI - Modulation of junctional communication by phosphorylation: protein phosphatases, the missing link in the chain. AB - Protein phosphorylation has been proposed to control the degree of intercellular gap junctional communication at several steps, from gene expression to protein degradation. In vertebrates, gap junctions are composed of proteins from the "connexin" (Cx) gene family, and the majority of connexins are post translationally modified by phosphorylation. Alterations in the phosphorylation status of proteins, resulting from the dynamic interplay of protein kinases and protein phosphatases, are thought to be involved in a broad variety of connexin processes (such as the trafficking, assembly/disassembly and degradation, as well as the gating of gap junction channels), but the underlying mechanisms remain poorly understood. Although protein kinases have an established role in this process (see Cruciani and Mikalsen, this issue), less is known about the involvement of protein phosphatases. The present review examines the role played by protein dephosphorylation catalysers in the regulation of gap junctional communication. PMID- 12566218 TI - Connexins, gap junctional intercellular communication and kinases. AB - A number of kinases and signal transduction pathways are known to affect gap junctional intercellular communication and/or phosphorylation of connexins. Most of the information is available for protein kinase A, protein kinase C, mitogen activated protein kinase, and the tyrosine kinase Src. Much less is known for protein kinase G, Ca(2+)-calmodulin dependent protein kinase, and casein kinase. However, the present lack of knowledge is not necessarily synonymous with lack of importance in the regulation of intercellular communication and phosphorylation of connexins. Kinases and the phosphorylation of connexins may be involved in the regulation of gap junctional intercellular communication at all levels ranging from the expression of connexin genes to the degradation of the gap junction channels. The exact role of the phosphorylation depends both on the kinase and the connexin involved, as well as the cellular context. PMID- 12566219 TI - Interaction of connexins with protein partners in the control of channel turnover and gating. PMID- 12566220 TI - Gap junctions and connexin expression in the normal and pathological central nervous system. AB - Gap junctions are widely expressed in the various cell types of the central nervous system. These specialized membrane intercellular junctions provide the morphological support for direct electrical and biochemical communication between adjacent cells. This intercellular coupling is controlled by neurotransmitters and other endogenous compounds produced and released in basal as well as in pathological situations. Changes in the expression and the function of connexins are associated with number of brain pathologies and lesions suggesting that they could contribute to the expansion of brain damages. The purpose of this review is to summarize data presently available concerning gap junctions and the expression and function of connexins in different cell types of the central nervous system and to present their physiopathological relevance in three major brain dysfunctions: inflammation, epilepsy and ischemia. PMID- 12566221 TI - Connexins and secretion. AB - Connexin channels clustered at gap junctions are obligatory attributes of all macroscopic endocrine and exocrine glands investigated so far and also connect most types of cells which produce secretory products in other tissues. Increasing evidence indicates that connexins, and the cell-to-cell communications that these proteins permit, contribute to control the growth of secretory cells, their expression of specific genes and their differentiated function, including their characteristic ability to biosynthetize and release secretory products in a regulated manner. Since the previous reviews which have been published on this topic, several lines of evidence have been added in support of multiple regulatory roles of gland connexins. Here, we review this novel evidence, point to the many questions which are still open and discuss some interesting perspectives of the field. PMID- 12566222 TI - Connexins and cancer. AB - The hypothesis, that gap junctional intercellular communication plays a key role in carcinogenesis and more generally in growth control was formulated nearly 40 years ago. From this time, data accumulated, showing that this type of communication is frequently decreased or absent in cells treated with tumor promoting agents, among transformed cells or between transformed/tumor cells and normal cells. This observation has been made on various cell types and whatever their tissue and species origins, by using in vitro and in vivo models. It led to the general assumption that the inhibition of gap junctional intercellular communication may play a role in carcinogenesis at two levels: (1) during tumor promotion by favoring the clonal expansion of initiated cells and (2) after the phenotypic transformation of cells by preventing the diffusion of putative "normalizing" factors between tumor cells and surrounding normal cells. During the past decade, the discovery that gap junction proteins, the connexins (Cx), may act as tumour suppressors, by reverting the phenotype of transformed cells confirmed the idea that their lack of function would be actively involved in carcinogenesis. However, we still do not know precisely what are the molecular processes that gap junctional intercellular communication may regulate and still do have very few data concerning the gap junction situation in human cancers. All these aspects are presented from an historical point of view and discussed below. PMID- 12566223 TI - The electrophysiology of gap junctions and gap junction channels and their mathematical modelling. AB - In most tissues of vertebrates, gap junctions control the exchange of ions and small molecules between adjacent cells, thus co-ordinating the cellular activities. The application of the dual voltage-clamp method to cell pair preparations enables one to elucidate the electrical properties of gap junctions and gap junction channels. The conductive and kinetic data obtained at the multichannel and single channel level led to a generalised concept for the operation of gap junction channels. Based on the biological data gained in this way, a mathematical model has been developed. This model is versatile and allows to simulate the electrophysiological behaviour of different types of vertebrate gap junctions. PMID- 12566224 TI - Lacandonia granules are present in Ginkgo biloba cell nuclei. AB - Lacandonia schismatica is a rare flowering plant with the sex organs spatially inverted. Several aspects of its cell biology are now known. Interestingly, within the cell nucleus, the chromatin is reticulated and it is associated to a novel structure named Lacandonia granules, a very abundant ribonucleoprotein particle showing similarities to perichromatin and Balbiani ring granules, which are involved in nuclear mRNA metabolism. To see whether these particles are present in other plants, we study the nucleus of Ginkgo biloba, a non-flowering plant. Light, electron and atomic force microscopy show that the cell nuclei of G. biloba are reticulated. Ultrastructural analysis showed that in the nucleoplasm, abundant intranuclear particles 32 nm in diameter are present. The EDTA regressive staining suggested that they contain RNA. Ultrastructural in situ hybridization confirmed the presence of RNA in these particles. Therefore, we conclude that the nuclei of G. biloba are reticulated and contain Lacandonia granules. We suggest that these particles may also be present in other plants. PMID- 12566225 TI - A continuous lineage of rat adenohypophysis stromal cells: characterisation and effects on GH(3)B(6) prolactin-secreting cell behaviour. AB - Although some studies have shown a possible modulation of the stroma on the hormonal secretion, it is not clear as to what are the requirements for these cellular interactions. In the present work, a homogeneous and continuous lineage of rat adenohypophysis stromal cells (APS9 cells) obtained from rat adenohypophysis primary culture was established. Using immunocytochemical methods and electron microscopy, we have characterised APS9 cells as elongated fibroblastoid-like cells with intercellular contacts, expressing alpha-smooth muscle actin, type IV collagen and laminin. By biochemical procedures, higher amounts of chondroitin sulphate and heparan sulphate were found in the pericellular and extracellular compartments of APS9 cell culture. In order to evaluate the possible effects of APS9 cell on GH(3)B(6) prolactin-secreting cell survival and/or proliferation, we established co-culture and proliferation assays. When GH(3)B(6) cells were cultivated on APS9 cell substrate, they displayed an organisation of many cellular cords strongly attached and covering all the stromal cell area, establishing punctual interactions or extensive surface associations between adjacent cells. Prolactin immunoreactivity appeared to be more scattered throughout the cytoplasm and accumulated in its periphery. When plated on glass coverslips, on newborn rat skin fibroblasts, on murine haematopoietic bone marrow stroma cell line or on murine foetal liver stroma cell line, GH(3)B(6) cells changed their organisation and presented a decrease in cell number and adherence to the substrate. Our results showed that the APS9 cell/GH(3)B(6) cell interactions favour cell growth and probably PRL secretion, and raises questions about the specificity of different organs and/or animal species stromas on the hormone secretion. PMID- 12566226 TI - Transforming growth factor beta activates Rac1 and Cdc42Hs GTPases and the JNK pathway in skeletal muscle cells. AB - The transforming growth factor beta (TGFbeta) plays an important role in cell growth and differentiation. However, the intracellular signaling pathways through which TGFbeta inhibits skeletal myogenesis remain largely undefined. By measuring GTP-loading of Rho GTPases and the organization of the F-actin cytoskeleton and the plasma membrane, we analyzed the effect of TGFbeta addition on the activity of three GTPases, Rac1, Cdc42Hs and RhoA. We report that TGFbeta activates Rac1 and Cdc42Hs in skeletal muscle cells, two GTPases previously described to inhibit skeletal muscle cell differentiation whereas it inactivates RhoA, a positive regulator of myogenesis. We further show that TGFbeta activates the C-jun N terminal kinases (JNK) pathway in myoblastic cells through Rac1 and Cdc42Hs GTPases. We propose that the activation of Rho family proteins Rac1 and Cdc42Hs which subsequently regulate JNK activity participates in the inhibition of myogenesis by TGFbeta. PMID- 12566227 TI - InsP(3)-mediated calcium release induced by heterologous expression of total chicory Leaf RNA. AB - A calcium dependent-chloride current (I(ni)) was recorded in Xenopus oocytes injected with total RNA from chicory leaf tissues, following depolarization from 35 to +60 mV. However, the signal transduction mechanism mediating I(ni) is unknown. The development of this current was mimicked by intracellular injection of the second messenger InsP(3) in control (non-injected) oocytes. Moreover, InsP(3) injection after I(ni) rundown did not reinitiate the current. The same phenomenon was observed following a second injection into control oocytes. Measurement of InsP(3) production in injected oocytes showed a net increase in the InsP(3) level on depolarization. Moreover, extracellular application of caffeine (5 mM) significantly reduced the number of oocytes displaying I(ni). Also, extracellular application of U-73122, a potent PLC inhibitor, clearly reduced the occurrence of I(ni). These data provide the first evidence that the calcium homeostasis mechanism induced by heterologous expression of total RNA from chicory leaves involves the InsP(3) signaling pathway. PMID- 12566228 TI - How can a nurse intervention help people with newly diagnosed epilepsy? A qualitative study of patients' views. AB - The aim was to describe the patients' views of the challenges posed by a new diagnosis of epilepsy and their assessment of a nurse intervention. Neurologists in South-East England referred patients into the study. Following a trial of a nurse intervention a subgroup of patients were purposefully identified for in depth interviews. Transcriptions of tape-recorded interviews were analysed using qualitative methodology. We found that younger people with epilepsy seemed to experience more trouble with driving, jobs and managing their lives in the context of new epilepsy, while older people saw epilepsy as just another illness to cope with. Patients reported difficulty in remembering what their doctors told them which they attributed partly to lack of time available in the consultation. They valued the time, and the technique of probing with explanations used by the nurse. The nurse intervention was seen as useful in making sense of symptoms, tests, risk management, and driving regulations and in helping manage their medicine taking. We conclude that people with newly diagnosed epilepsy face different challenges, some of which are related to their age at diagnosis. Patients reported help from the nurse with understanding the diagnosis, tests, risk management and taking their medication. Follow-up is necessary to measure behavioural effects on self-management in the long-run. PMID- 12566229 TI - Prescribing and the epilepsy specialist nurse. AB - Increasingly, the epilepsy nurse specialist has become an integral part of the specialist epilepsy team. Nurse specialists, who practice at an advanced level, frequently advise patients on diagnosis and antiepileptic drug changes. The inclusion of antiepileptic drugs to the nurse Prescribing Formulary would allow specialist nurses to provide a more enhanced service to patients. PMID- 12566230 TI - Implementing good practice in epilepsy care. AB - Examples of evidence-based guidelines for epilepsy care exist. However, guidelines are of little use if they are not recognised, implemented and supported. The object of this study was to establish the degree to which good practice guidelines for epilepsy have been implemented and to identify positive and negative factors that affect their implementation. Semi-structured questionnaires were sent to 750 randomly selected health professionals working in primary and secondary care in England. The sample comprised nurses (200), adult consultants (including learning disability consultants) (300), paediatric consultants (150) and general practitioners (100). Aspects of good practice are being implemented in some areas, but not generally, therefore service provision is likely to remain fragmented until this is addressed. Professionals have been prevented from successful implementation of guidelines to sustain good practice due to a number of factors, most notably lack of time, workload, competing priorities and staffing levels. Factors that have promoted and encouraged the successful adoption and application of good practice include inputs from epilepsy specialist nurses (ESNs), appropriate, timely and accessible professional development opportunities and the support and enthusiasm of colleagues. PMID- 12566231 TI - Tele-EEG in epilepsy: review and initial experience with software to enable EEG review over a telephone link. PMID- 12566232 TI - Electrophysiological, neurochemical and regional effects of levetiracetam in the rat pilocarpine model of temporal lobe epilepsy. AB - This study compared levetiracetam (Keppra) with reference antiepileptic drugs (AEDs) in the rat pilocarpine model of temporal lobe epilepsy. Electroencephalogram (EEG) recordings showed that i.p. administration of valproate (300 mg/kg), phenobarbital (5 mg/kg) and clonazepam (0.5 mg/kg) all significantly delayed the appearance of the first epileptic spike discharge in hippocampus as well as synchronous epileptiform activity in hippocampus and cortex. In contrast, i.p. administration of levetiracetam (17 mg/kg) only significantly delayed the appearance of the latter. This was corroborated by findings showing that i.p. administration of levetiracetam (17 mg/kg) significantly opposed pilocarpine-induced increases in the amplitude of the orthodromic population spike in the hippocampal CA3 area of urethane anaesthetised rats, while valproate (200 mg/kg), phenobarbital (10 mg/kg) and clonazepam (1 mg/kg) had no effect. Pre-treatment i.p. with phenobarbital (10 mg/kg) and clonazepam (0.5 mg/kg) significantly reversed seizure-induced changes in aspartate and GABA concentrations while valproate (300 mg/kg) significantly reduced aspartate concentrations further. In contrast, levetiracetam (34 mg/kg) significantly counteracted all seizure-induced alterations in amino acid concentrations. Midazolam induced significant seizure protection after microinjection into substantia nigra pars reticulata (SNR, 50 nmol), nucleus accumbens (NA, 25 nmol) and caudate putamen (CP, 25 nmol), whereas phenytoin (50 nmol) only showed significant seizure protection after injection into the latter area. Levetiracetam differed by significant seizure protection after injection into SNR (1,000 nmol) and NA (3,000 nmol). These results suggest that levetiracetam is distinct from other AEDs by its ability to selectively suppress synchronisation of neuronal spike and burst firing in hippocampus. PMID- 12566233 TI - Psychosocial reactions to the epilepsy in an unselected group of patients with epilepsy under treatment in general hospitals. AB - OBJECTIVE: This study aimed at exploring the psychosocial reactions to epilepsy in a group of patients with uncomplicated well-controlled epilepsy. METHOD: One hundred and ninety-eight patients, referred by neurologists in a general hospital completed a questionnaire, consisting of simple open questions about the epilepsy and the drug treatment. Additionally, the patients were asked to complete the QOLIE-10-scale as well as a short questionnaire with questions about attitudes towards epilepsy and the treatment. RESULTS: The overall psychosocial outcome was favourable in our study sample. About 50% of the patients have a paid job, have an average educational level and the majority (almost 70%) of the patients have a partner. This is not different from findings in the general population. In line with these social findings, the patients report in majority that the epilepsy is well integrated: as a rule the family and friend are informed and they feel that the epilepsy is accepted in society. Restrictions are limited to activities such as swimming. Consequently, quality of life (QOL) is high for the majority of the patients, with only 6% of the patients reporting a low QOL. DISCUSSION: It is often claimed that people with epilepsy, as a group, have more psychosocial problems than control populations of healthy subjects. As this may be true for the average comparisons between patients with epilepsy and controls, we must take into account during clinical practice that these problems occur predominantly in the minority of patients with refractory epilepsy. Patients with well-controlled epilepsy do not show worrying reactions to the epilepsy. PMID- 12566234 TI - Investigation of probable relationship between Toxoplasma gondii and cryptogenic epilepsy. AB - PURPOSE: Cryptogenic epilepsy, defines a group of epilepsy syndromes for which an aetiology is unknown but an underlying brain disease is suspected. We selected patients in this subgroup of epilepsy and investigated the sero-positivity rate for anti-Toxoplasma IgG antibodies by Enzyme Linked Immunosorbent Assay (ELISA). We investigated the probable relationship between Toxoplasma gondii and cryptogenic epilepsy. METHODS: We selected 50 patients with cryptogenic epilepsy, 50 patients with known cause epilepsy and 50 healthy volunteers and investigated the sero-positivity rate for anti-Toxoplasma IgG antibodies by ELISA. RESULTS: The sero-positivity rate for anti-Toxoplasma IgG antibodies among cryptogenic epilepsy patients (52%) was found to be higher than healthy volunteers (18%) and known cause epilepsy patients (22%) with statistical significance, (X(2)=18.095, P<0.01). CONCLUSION: There might be a causal relationship between chronic toxoplasmosis and the aetiology of cryptogenic epilepsy. PMID- 12566235 TI - Epilepsy in children with cerebral palsy. AB - OBJECTIVES: To study the occurrence, associated factors, nature and prognosis of seizures in children with cerebral palsy (CP). DESIGN: A prospective, descriptive, hospital-based, case-control study. SETTING: Tertiary level University Teaching Hospitals in the Al Ain Medical District, United Arab Emirates. PATIENTS: Fifty-six children with CP and seizures seen in the neurodevelopmental clinics at Al Ain and Tawam University Hospitals during the period of 1997-1999 were studied (group 1). Two control groups of 35 children with CP without seizures (group 2) and 50 children with seizures but no CP (group 3) were also studied. RESULTS: Spastic tetraplegia was the commonest type of CP associated with seizures whereas spastic diplegia was the commonest variety of CP in group 2. Most children with CP had an early onset of seizures within the first year of life as against those without CP. The children in group 1 had a higher incidence of neonatal seizures (42.9% vs. 29.4% in group 2 and 0% in group 3), presence of significant developmental delay (98.2% vs. 20.0% in group 3), occurrence of significant abnormalities on brain imaging (94.6% vs. 19.6% in group 3) and a need for use of more than 1 antiepileptic drug (66.1% vs. 30.0% in group 3). Over half of children in the study group presented with generalized tonic clonic seizures; the electroencephalogram (EEG) showed focal epileptic discharges with or without secondary generalization in 39.3%. The overall outcome of seizures in children with CP was poor needing prolonged course of anticonvulsant medications, polytherapy and higher incidence of refractory seizures and admissions for status epilepticus compared to the control group. CONCLUSIONS: Cerebral palsy is associated with a higher incidence of seizure disorders, which, in a majority, has its onset in the neonatal period; brain imaging showed abnormal pathology in most affected children, which possibly accounts for the tendency to more refractory seizures in these children. PMID- 12566236 TI - Seizure-alert dogs: a review and preliminary study. AB - PURPOSE: Gather data on incidence of canine alerting/responding behavior with a defined patient population. Research development and use of purported alerting dogs. METHODS: Review of the literature was performed. A qualitative questionnaire was completed by epilepsy patients. Service dog trainers were identified. RESULTS: Of 63 patients, 29 owned pet dogs. Nine reported their dog responded to seizures, three also were reported to alert to seizure onset. There was no significant evidence of correlation between alerting/responding behavior and the patients' demographics, health, or attitude/opinion of pets. Seizure alerting/responding behavior of the dog did not appear to depend on its age, gender or breed. A literature review revealed psychological and practical benefits of service dogs are well documented. Fifteen trainers of seizure-assist dogs were identified and interviewed. CONCLUSIONS: Findings suggest some dogs have innate ability to alert and/or respond to seizures. Suggests a trend in type of seizure/auras a dog may alert to. Success of these dogs depends largely on the handler's awareness and response to the dog's alerting behavior. Warrants further research to aid in the selection of patients who may benefit from seizure-assist dogs, for identification and further training of these dogs and possibly the development of seizure-alerting devices. PMID- 12566237 TI - Risk factors and outcome of mood disorders in epilepsy: a case-control study. AB - BACKGROUND: This case-control study investigated both the risk factors and outcome of mood disorders in epilepsy. METHODS: For this study, 44 patients with both epilepsy and a mood disorder (study group) were compared with 44 randomly selected patients of epilepsy without a mood disorder (control group). Psychiatric diagnosis was made as per ICD-10 Diagnostic Criteria for Research (ICD-10 DCR). International classification for seizure types (1981) was used for classification of seizure types. RESULTS: Of the patients in the study group, a majority were educated up to at least primary level, had later onset of seizures, longer duration of epilepsy and cluster attacks. The outcome of mood disorders in epilepsy was found good in most. CONCLUSIONS: Educated patients who develop epilepsy at a later age and patients with poorly controlled epilepsy are more likely to experience mood disorders. In most patients with epilepsy, mood disorders remit completely; notably, in some patients affective symptoms resolve spontaneously. PMID- 12566238 TI - Subacute encephalopathy with seizures in alcoholism. AB - Generalised seizures are a common manifestation of acute alcohol intoxication. Alcohol is known to precipitate generalised seizures in patients with focal brain pathology. A rare case of secondarily generalised seizures precipitated by alcohol in a patient without an underlying focal brain lesion is reported. Electroencephalopgraphy (EEG) showed periodic lateralised epileptiform discharges (PLEDs) on the side contralateral to the focal motor fits. PMID- 12566239 TI - Selective cholesteryl ester uptake from high density lipoprotein by human first trimester and term villous trophoblast cells. AB - As villous trophoblast does represent the contact zone between foetal and maternal tissues, the present in vitro study was aimed at investigating cholesterol supply from human high density lipoprotein subclass 3 (HDL(3)) to trophoblast cells isolated from human first trimester and term placenta. Binding of (125)I-HDL(3) was specific and saturable with similar K(d)-values for first trimester (54 microg HDL(3)-protein/ml) and term villous trophoblast cells (29 microg HDL(3)-protein/ml). The cell-association of (125)I-HDL(3) was 3-fold higher for term trophoblast cells while the specific cell-association of [(3)H]cholesterol ester(CE)-labelled HDL(3) was higher for first trimester trophoblast preparations. As a consequence, first trimester trophoblast cells have a pronounced capacity for selective CE-uptake from HDL(3). Competition experiments with native and oxidized low-density lipoprotein as well as cAMP mediated stimulation of cell-association of [(3)H]CE-HDL(3) in both trophoblast preparations suggested the scavenger receptor class B, type I (SR-BI) as a likely receptor mediating this pathway. SR-BI m RNA could be identified by RT-PCR and Northern blot experiments in both trophoblast preparations. Western blot analysis and immunocytochemistry revealed high expression of SR-BI in first trimester trophoblast. A polyclonal antiserum raised against murine SR-BI significantly decreased cell-association of [(3)H]CE-HDL(3) in trophoblast cells. We conclude that human first trimester and term trophoblast cells express SR-BI which could serve as an efficient route for supplying cholesterol esters from maternal lipoproteins to foetal tissues. PMID- 12566240 TI - Expression of organic anion transporting polypeptide E (OATP-E) in human placenta. AB - PURPOSE: To examine the expression of multifunctional Na(+)-independent organic anion transporting polypeptide, termed OATP-E, involved in the transport of thyroid hormone in human placenta. METHODS: Western blot analysis was performed using a specific antibody against OATP-E in human placenta to confirm the expression of OATP-E at the protein level. Immunohistochemistry was also performed using the specific antibody against OATP-E on frozen sections of human placenta. RESULTS: By Western blot analysis, a single band for OATP-E was observed in human placenta. Immunohistochemistry revealed that OATP-E was predominantly expressed at the apical surface of the syncytiotrophoblasts in placenta. CONCLUSION: These results reveal that OATP-E is expressed in human placenta, suggesting a functional role for the transplacental transfer of thyroid hormone. PMID- 12566241 TI - Binding of acetaldehyde to human and Guinea pig placentae in vitro. AB - BACKGROUND: Significant interindividual variability exists following maternal alcohol consumption; not all children born to alcoholic women manifest the symptoms associated with foetal alcohol spectrum disorder (FASD). OBJECTIVE: To investigate the potential role of the placenta as a source of variability by determining if interindividual variability exists in the binding of acetaldehyde to human placenta. METHODS: Acetaldehyde was added to ten different human placental homogenates and subjected to equilibrium dialysis. Homogenates of placentae obtained from guinea pigs chronically exposed to ethanol throughout gestation were also dialysed in the presence of acetaldehyde to look for alterations in binding after chronic alcohol exposure. Nonlinear least-squares regression analysis was used to characterize the binding system involved. RESULTS: It was found that the amount of acetaldehyde bound to human placentae varied by as much as 3-fold among placentae. The binding profile of acetaldehyde was characterized as a two site binding system (Ka(1)=9.8 x 10(5)+/-0.7 x 10(5)l/mol, N(1)=1.1 x 10(-8)+/-0.7 x 10(-8)mol/g tissue; Ka(2)=1.6 x 10(4)+/-0.9 x 10(4)l/mol, N(2)=1.7 x 10(-7)+/-0.4 x 10(-7)mol/g tissue). Chronic alcohol exposure had no effect on the degree of acetaldehyde binding. CONCLUSION: This previously unidentified source of variability may partially explain why some foetuses are adversely affected by prenatal alcohol exposure while others are not. PMID- 12566242 TI - Regulation of trophoblastic gelatinases by proto-oncogenes. AB - During the first trimester of pregnancy, certain cytotrophoblastic cells (CTB) of anchoring villi invade the underlying decidua. Regulation of this invasive behaviour depends on cytokines and growth factors secreted by decidua and trophoblast, which modulate metalloproteinase (MMP) secretion of CTB. Since MMP-9 expression by CTB is a prerequisite for matrigel invasion and since the promoter region of the MMP-9 gene contains two AP-1 binding sites, we hypothesized, that transient activation of c-jun and c-fos oncogenes (which bind to form AP-1) by tumour necrosis factor (TNFalpha), or the phorbol ester TPA will promote the invasive phenotype of CTB and induce the production of MMP-9.TNFalpha or TPA when added to primary cultures of CTB increase MMP-9 activity and MMP-9 mRNA. This effect is inhibited by cycloheximide indicating the necessity of protein synthesis. TPA or TNFalpha induces also the binding of nuclear proteins (extracted from treated CTB) to a radiolabelled oligonucleotide corresponding to the consensus sequence of the TPA responsive element. Antibodies to Jun and Fos can displace this binding. Transient transfection of antisense mRNA to jun or fos into CTB inhibits the immunoreactivity and gelatinolytic activity of MMP-9. We conclude that AP-1 is necessary but may not be sufficient for transactivation of the MMP-9 gene in human CTB. PMID- 12566243 TI - Expression and activation of MAP kinases, ERK1/2, in the human villous trophoblasts. AB - Mitogen-activated protein kinase (MAP kinase) plays a central role in the signal transduction for diverse cellular responses, such as proliferation, differentiation, stress response and cell death, via activation after binding of growth factors to the respective receptors on the cell membrane. In the human placental tissues, however, little is known about the expression and activation of the classical MAP kinases, extracellular signal-regulated kinase1/2 (ERK1/2). We therefore examined the expression of ERK1/2 in the human chorionic and placental tissues between 5 and 41 weeks of gestation, using Western blotting, immunohistochemistry and in situ hybridization. To explore the activation of ERK1/2 protein, we used an antibody that reacts with both phosphorylated and non phosphorylated ERK1/2 (total ERK1/2), as well as antibodies that react only with phosphorylated ERK1/2. The expression pattern of phosphorylated ERK1/2 in the trophoblasts was compared with that of various growth factor receptors, such as c met, IGF-1R, flt-1, EGFR, PDGFR, Bek, and flg. Total ERK1/2 was immunolocalized in the villous cytotrophoblasts (CTs), but not in the syncytiotrophoblasts (STs), throughout pregnancy. In situ hybridization also showed the localization of ERK1 mRNA in the villous CTs. Interestingly, however, phosphorylated ERK1/2 was immunolocalized in the villous CTs only up to 12 weeks of gestation. Western blot also showed the stronger bands of phosphorylated ERK1/2 in the tissues of the first trimester. Among the growth factor receptors, c-met was strongly expressed in the villous CTs during the first trimester, and resembled the expression pattern of phosphorylated ERK1/2. These findings suggest that the MAP kinase pathway is activated in the villous CTs during the first trimester in the human placenta. PMID- 12566244 TI - Physiological apoptotic agents have different effects upon human amnion epithelial and mesenchymal cells. AB - Foetal membrane rupture is thought to follow from gene-controlled tissue remodelling and apoptosis. We reported previously that staurosporine, cycloheximide, actinomycin D, as well as more physiological apoptotic agents (lactosylceramide, 15d-PGJ(2)) increase prostaglandin release in parallel with induction of apoptosis in WISH and amnion epithelial cells. Also, inhibition of prostaglandin release by cyclooxygenase inhibitors or PKA activators is accompanied by a parallel decrease in apoptosis. We hypothesize that amnion prostaglandin metabolism is linked with apoptosis in amnion epithelial cells and thus to membrane rupture. Amnion mesenchymal cells are also critical for membrane integrity. Their susceptibility to apoptotic agents is unknown and is the subject of this report. In amnion epithelial cells, lactosylceramide (125 microM) induced 6.5-fold, 20-fold increases in PGE(2) and NMP production (apoptosis), respectively. Conversely, in mesenchymal cells, lactosylceramide doses up to 200 microM had no effect on PGE(2) or NMP release. In both cell types, incubation with 15d-PGJ(2) (5-100 microM) demonstrated dose and time dependent increases in PGE(2) and NMP. PKA activators inhibited 15d-PGJ(2) induced PGE(2) release and apoptotis in epithelial cells, but not in mesenchymal cells, however. Major amnion cell types have different sensitivities to physiological apoptotic agents. Prostaglandin release occurs coincident with apoptosis in both amnion epithelial and mesenchymal cells. PMID- 12566245 TI - Hypoxia favours necrotic versus apoptotic shedding of placental syncytiotrophoblast into the maternal circulation. AB - In the third trimester of normal pregnancy, the mother tolerates daily shedding of several grams of dying placental trophoblast into the maternal circulation. The balance between apoptotic and necrotic shedding is presently unknown. Since pre-eclampsia is characterized by an altered placental oxygenation and increased trophoblast shedding, we investigated the role of oxygen on the balance of apoptotic versus necrotic trophoblast shedding in vitro. We studied human trophoblast turnover in explanted villi from late first and third trimester placentas in low oxygen (2 per cent) and higher oxygen tensions (6 per cent and 18 per cent) for up to 72h. Trophoblast turnover including apoptosis and necrosis were assessed by histology, immunolocalization of Mib-1 (proliferation marker), Bcl-2 (apoptosis inhibitor), activated caspase 3 (apoptosis promoter), cytokeratin 18 neo-epitope formation (M30 antibody), TUNEL test (DNA degradation), and (3)H-cytidine and(3) H-uridine incorporations. Culture in 2 per cent oxygen increased cytotrophoblast proliferation and syncytiotrophoblast shedding by necrosis. The proteins necessary for execution of apoptosis were mostly retained in the cytotrophoblast due to lack of syncytial fusion. Culture in 6 per cent and 18 per cent oxygen reduced cytotrophoblast proliferation. Syncytial fusion occurred and activity of caspase 3 was found in the syncytiotrophoblast; the latter remained intact demonstrating physiologic turnover, including apoptotic shedding. We conclude that severe placental hypoxia favours necrotic rather than apoptotic shedding of syncytial fragments into the maternal circulation. Since uteroplacental ischaemia is a significant risk factor for pre-eclampsia, these findings may explain the link between reduced uteroplacental blood flow and the systemic clinical manifestations of this disease. PMID- 12566246 TI - Changes in fetal capillaries during preplacental hypoxia: growth, shape remodelling and villous capillarization in placentae from high-altitude pregnancies. AB - Patterns of fetoplacental angiogenesis and villous growth vary in pregnancies complicated by different forms of fetal hypoxia. This study uses stereological estimators to examine absolute and relative dimensions of villi and fetal capillaries in cases of preplacental hypoxia due to pregnancy at high altitude. Placental samples were drawn from well-defined subjects in different ethnic groups born, raised and completing term pregnancies at low (500 m) and high altitude (3600 m above sea level). Volumes, surfaces and lengths were used to monitor the nett growth of villi and capillaries. Indices of villous capillarization comprised volume, surface and length densities and capillary:villus surface and length ratios. Villus/capillary 'calibre' and shape were quantified using cross-sectional areas, perimeters and shape coefficients (perimeter(2)/area). Group comparisons were drawn by two-way analyses of variance with altitude and ethnicity as the main factors. Volumes, surfaces and lengths of villi, and volumes of capillaries, were reduced at high altitude. Capillary volume declined primarily by formation of narrower microvessels which were more irregular in outline. There were no differences in capillary surface area or length. Cross-sectional sizes and shapes of villi were unaltered. Differences in villous capillarization were confined to higher surface and length densities. Ethnic differences in villous length, capillary length and cross-sectional area tended to favour native groups who are pre-adapted to life at high altitude. Results show that high-altitude pregnancy is not accompanied by increased angiogenesis but may involve enhanced villous capillarization and vascular shape remodelling. Comparisons are drawn with changes seen in maternal anaemia. It is concluded that absolute and relative measures of villous and capillary growth are required lest misinterpretations are introduced by equating hypercapillarization with enhanced angiogenesis or the pattern of capillary branching. The importance of controlling for various potential confounders is also emphasized. PMID- 12566247 TI - Impaired oxygen-dependent reduction of HIF-1alpha and -2alpha proteins in pre eclamptic placentae. AB - Pre-eclamptic (PE) placentae overexpress hypoxia inducible transcription factors 1alpha and -2alpha proteins (Biol. Repro. 64: 499-506, 2001; Ibid 1019-1020). Possible explanations include (a) impaired oxygen-dependent reduction, and/or (b) enhanced sensitivity to reduced oxygen. After 18 h equilibration under 21 per cent O(2) atmosphere, we subjected villous explants prepared from placentae of normal pregnant (NP) and pre-eclamptic (PE) women (n=8 each) to 4h of hypoxia (2 per cent oxygen), and then studied the disappearance of HIF-1alpha and -2alpha proteins during subsequent oxygenation over 90 min (21 per cent oxygen). The disappearance of these HIF proteins as assessed by Western analysis was significantly impaired in the pre-eclamptic tissues. Even after 18h equilibration under a 21 per cent O(2) atmosphere, and then a further 4h at 21 per cent O(2), HIF-1alpha and -2alpha protein expression remained increased in villous explants from PE women (both P< 0.04 vs NP). To address whether chronic hypoxia per se (which is believed to exist in the pre-eclamptic placenta) might contribute to these findings, we subjected villous explants from normal placentae (n=6) to 18 h preincubation under 2 per cent or 21 per cent oxygen prior to subsequent incubation for 4h at 2 per cent oxygen and then 90 min at 21 per cent oxygen. The time course of disappearance of HIF proteins during oxygenation was similar irrespective of the 2 per cent or 21 per cent preconditioning. To evaluate oxygen sensitivity, we exposed villous explants from NP and PE women (n=6 each) to different oxygen atmospheres for 4h and measured HIF protein induction. Although the data showed a significant inverse relationship between HIF expression and oxygen concentration, there was no significant difference between the slopes of this relationship for the two groups of women. We conclude that villous explants from PE placentae fail to adequately downregulate HIF protein expression upon oxygenation. This abnormality may contribute to their overexpression in vivo. PMID- 12566248 TI - Identification of multiple differentially expressed messenger RNAs in normal and pathological trophoblast. AB - In an attempt to assess the molecular basis of phenotypic alterations present in the gestational trophoblastic diseases (GTDs) and to identify genes whose expression is specifically associated to these placental proliferative disorders we performed differential display techniques. Initially 19 candidate gene fragments were identified and differential expression was confirmed in eight of these fragments by Northern blot analysis. At the mRNA level ribosomal L26 (rL26), ribosomal L27 (rL27), a new Kruppel type zinc finger protein and TIS11d were preferentially expressed in normal early placenta (NEP) relative to complete hydatidiform mole (CHM), persistent gestational trophoblastic disease (PGTD) and choriocarcinoma JEG-3 cell line. In contrast, heterogeneous ribonucleoprotein A1 (hnRNPA1), the ferritin light chain mRNA, and the uncharacterized protein KIAA0992 were predominantly expressed in JEG-3 cell line. Finally, decorin, a prototype member of an expanding family of small leucine-rich proteoglycans, showed high expression in CHM. In addition we demonstrated by immunohistochemistry analysis that increased decorin mRNA in CHM reflected a genuine augmentation in average steady state mRNA levels within cells. Taken together, these findings provide several interesting candidates for regulation of tumorigenic expression as well as early placentation development, including those involved in protein synthesis (rL26 and rL27), metabolism (ferritin light chain), intercellular communication (decorin) and regulation of gene expression (Kruppel like zinc finger, TIS11d and hnRNPA1). Information about such alterations in gene expression could be useful for elucidating the genetic events associated to gestational trophoblastic pathogenesis, developing new diagnostic markers, or determining novel therapeutic targets. PMID- 12566249 TI - Stereological investigation of placental morphology in pregnancies complicated by pre-eclampsia with and without intrauterine growth restriction. AB - The aim of this study was to quantify placental morphology in pregnancies complicated by pre-eclampsia with and without intrauterine growth restriction. Particular attention is given to the dimensions and composition of peripheral (intermediate+terminal) villi. Placentae from 9 control pregnancies, 5 cases of pre-eclampsia, 5 cases of intrauterine growth restriction and 5 cases of pre eclampsia with intrauterine growth restriction were randomly sampled for location and position. Formalin-fixed, wax-embedded sections stained by the Masson trichrome method were subjected to stereological analysis in order to quantify the volumes of placental components and the surfaces and derived diameters for peripheral villi and fetal capillaries. Group comparisons were drawn using two way analysis of variance. Fetal weights were reduced in all complicated pregnancies but only intrauterine growth restriction was accompanied by a significantly smaller placenta. Pre-eclampsia was not associated with main effects on placental morphology and (except for trophoblast thickness) there were no interaction effects involving pre-eclampsia. In contrast, intrauterine growth restriction was associated with a placenta which had reduced volumes of intervillous space and all types of villi (stem, intermediate, terminal). The impoverished growth of peripheral villi affected all tissues (trophoblast, stroma, capillaries) and was accompanied by smaller exchange surface areas and a thicker trophoblastic epithelium. The derived mean diameters of villi and capillaries were not affected. It is concluded that intrauterine growth restriction, but not pre-eclampsia, is associated with substantial changes in placental morphology including impoverished growth of villi and fetal vasculature. These changes are likely to reduce placental oxygen diffusive conductances and contribute to fetal hypoxic stress. PMID- 12566250 TI - Metabolite concentrations in human term placentae and their changes due to delayed collection after delivery. AB - The present study was designed to quantify the major cellular metabolites in human placentae and their changes due to the confounding effect of time and subsequent hypoxia during sample collection using magnetic resonance spectroscopy ((1)H-and (31)P-MRS). The absolute placental concentrations of lactate, glucose, major amino acids and cellular volume/osmo-regulators, glutathione, high-energy phosphates, fatty acids, phospholipids, triglycerols, and cholesterol are reported. There were no spatial differences in metabolism or protein expression throughout the placenta. The most significant temporal changes, due to the collection time (from 1 to 25 min after delivery), were increased concentrations of lactate (r=0.996, statistically significant P< 0.01 after 11 min) and decreased concentration of glucose and ATP (r=-0.963 and -0.97, respectively, P< 0.01 after 11 min). The placental samples from the later collection groups (16-24 min) had also significantly lower level of NAD(+) (r=-0.95, P< 0.01). Only the latest collection group (21-24 min) had increased lipid peroxidation and changes in lipid metabolites (P< 0.01). We conclude that the optimal window for collecting placental tissue without incurring metabolic artifacts due to hypoxic conditions is within 9 min of placental delivery. PMID- 12566251 TI - The effects of maternal nutrition and body condition on placental and foetal growth in the ewe. AB - This study investigated the effects of maternal body condition and nutrition on placental and foetal growth in mid-gestation. Welsh Mountain ewes (n=24) of body condition 3.5 (high, H) and 2.0 (low, L) at mating, were fed either 100 per cent or 70 per cent of their daily maintenance requirements from day 22 of gestation, yielding four groups: H100 (n=5), H70 (n=6), L100 (n=7) and L70 (n=6). On day 65, placental and foetal parameters were measured. Whilst the placentome number tended to be lower in L than H ewes, the mean placentome weight was significantly greater in L100 than H100 animals. Nutritionally related changes in IGFBP expression within the placentome and intercotyledonary endometrium may explain these findings, with IGFBP-3 expression in the luminal epithelium and caruncular stroma of the placentome villi being inversely correlated to placentome number and the total placentome weight respectively. The foetal CRL was shorter and the ponderal index greater in L than H ewes. The foetal CRL was positively correlated to maternal IGF-I concentrations and the placentome number, although the foetal weight remained unaltered by treatment. This study therefore demonstrates that body condition and ration can alter foetal and placental growth, perhaps by modifying systemic parameters and uterine IGF expression. PMID- 12566252 TI - Effect of maternal overnutrition during pregnancy on pituitary gonadotrophin gene expression and gonadal morphology in female and male foetal sheep at day 103 of gestation. AB - The aim was to determine whether nutritionally mediated restriction of placental growth alters foetal body growth, pituitary gonadotrophin gene expression and gonadal development at Day 103 of gestation. Embryos recovered from adult ewes inseminated by a single sire were transferred, singly, into the uteri of adolescent recipients. After transfer, adolescent ewes were offered a high (H, n=16) or moderate (M, n=12) level of a complete diet. Ewes were slaughtered at 103+/-0.2 days of gestation and foetal blood, brain, pituitary and gonads were collected. Mean placental weight was lower (P< 0.01) in H than in M groups but foetal weight and reproductive organ weights were similar. Maternal nutrition did not influence LHbeta or FSHbeta mRNA expression in either sex but FSHbeta mRNA expression was higher (P< 0.001) in female (n=11) than in male (n=17) foetal pituitaries. Mean foetal plasma gonadotrophin concentrations were not influenced by dietary intake in either sex. Plasma progesterone concentrations were lower (P=0.001) in foetuses derived from H compared with M intake dams. Compared with M foetuses (n=5), ovaries from H foetuses (n=6) had fewer primordial follicles (P< 0.05) and fewer follicles in total (P< 0.005). In contrast, maternal nutritional status did not influence either seminiferous cord or Sertoli cell numbers in male foetuses (H, n=10; M, n=7). It is concluded that high maternal nutrient intakes restricted placental growth and altered foetal ovarian follicular development prior to the end of the second third of gestation. The latter effect was independent of gonadotrophin secretion.Crown PMID- 12566253 TI - A three-dimensional cell culture model for bovine endometrium: regeneration of a multicellular spheroid using ascorbate. AB - The development of a multicellular spheroid comprising bovine endometrial epithelial cells (BEE) and bovine endometrial stromal cells (BES) is described in this study. The BES were cultured to confluence in medium with L -ascorbic acid phosphate magnesium salt n -hydrate (AsA-P) which stimulates collagen synthesis in BES. The BEE were co-cultured on a BES cell-sheet for 24h before detachment of the cell-sheet to generate a hetero-spheroid. After EDTA treatment and agitating with pipette, the floating cell-sheet shrank and became an aggregated cell mass in a few days; it finally formed a round-shaped hetero-spheroid composed of BES and BEE. Histological examination found that hetero-spheroids were covered with BEE on the outer layer. When cell viability was examined with TUNEL (terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling), no positive signal was detected in the spheroid for up to 10 days. Immunofluorescence observations showed that spheroids contained abundant extracellular matrices, including type-I, -III, -IV collagen, fibronectin, and laminin. PGF(2alpha) produced by hetero-spheroids in response to oxytocin was significantly higher than those produced by monolayer cultured BEE (P< 0.05). MMPs were not detected in media from spheroids cultured for 5 days after detachment of the cell sheet. These results indicate that bovine endometrial cells have the capacity to regenerate as a multicellular spheroid after treatment with ascorbate in vitro. The spheroid displays an endometrium-mimic feature. Thus, we conclude that spheroids formed by BES and BEE are a useful in vitro model of bovine endometrium. PMID- 12566254 TI - X-inactivation patterns in human embryonic and extra-embryonic tissues. AB - Mice have skewed X chromosome inactivation (XCI) in extraembryonic tissue while examination of human placentae have yielded conflicting results. We investigated XCI patterns in human embryonic and extra-embryonic tissues. First and early second trimester placental and foetal tissues were collected. Cytotrophoblasts were isolated from the placentae. Female samples were identified and X inactivation patterns were determined by analysis of androgen receptor (HAR) methylation patterns. Among 55 females heterozygous at the HAR, 37 had random and 18 skewed XCI. In foetal tissues a skewed XCI pattern was only observed in one liver and one intestine sample. A greater incidence of skewed XCI pattern was present in extra-embryonic compared to embryonic tissues (P=0.022). A markedly skewed XCI pattern was only found in one cytotrophoblast sample. Random and skewed XCI patterns were detected in human embryonic and extra-embryonic tissues. The extra-embryonic tissue had a higher proportion of skewed XCI, but marked skewed XCI was uncommon in both tissues. Skewed XCI may not play a role in normal human placentation. PMID- 12566255 TI - Contribution of the umbilical cord and membranes to untrimmed placental weight. AB - The extent of placental trimming before weighing varies within and between obstetric units. Quantification of the contribution of the umbilical cord and membranes to placental weight is required to enable measurements to be compared across populations. In a sample of 50 neonates born in Southampton, placentae of liveborn singletons were weighed three times; after removing any obvious blood clots, after cutting the umbilical cord, and after stripping both the foetal and maternal membranes. The correlation between untrimmed and trimmed placental weight was 0.98. Since the combined weight of the cord and membranes increased with increasing trimmed weight (correlation=0.54), the percentage rather than absolute difference between untrimmed and trimmed weight was calculated. The median difference between untrimmed and trimmed weight was 16 per cent. No association was found between the percentage difference and infant sex, duration of gestation, birthweight, maternal age, labour onset and duration, and presentation at delivery. However, the mode of delivery had a significant effect on this difference; medians for vaginal and Caesarean section deliveries were 19 per cent and 14 per cent respectively. PMID- 12566256 TI - Abruptio placentae and highest maternal serum activin A levels at mid-gestation: a two cases report. PMID- 12566257 TI - Chilling-dependent photoinhibition, nutrition and growth analysis of Eucalyptus nitens seedlings during establishment. AB - Effects of chilling-dependent photoinhibition on gas exchange, chlorophyll fluorescence, growth and nutrition of Eucalyptus nitens (Deane and Maiden) Maiden seedlings were assessed for 70 weeks after transplanting 9-month-old seedlings in early winter. One month before transplanting, the seedlings were assigned to fertilized or nutrient-deprived treatments. Immediately after transplanting, half the seedlings in each nutrient treatment were placed in shadecloth tree shelters. The experimental site was at an altitude of 700 m, which is considered marginal for the establishment of E. nitens plantations in Tasmania because of low mean annual minimum temperatures. Overnight frosts followed by sunny morning conditions in the first 20 weeks after transplanting (early June to early October) caused severe photoinhibition. Predawn maximal photochemical efficiency (Fv/Fm) and maximum net photosynthesis (Amax) were depressed in nutrient-deprived seedlings compared with fertilized seedlings, although shading partially alleviated this difference. Neither Fv/Fm nor Amax recovered to values observed before transplanting until > 20 weeks after transplanting. During this period, non-photochemical quenching (NPQ) was high in seedlings in all treatments, although NPQ was lower in shaded, fertilized seedlings than in seedlings in the other treatments. Total foliar nitrogen (N) concentration increased up to 42 weeks after transplanting in the nutrient-deprived seedlings in parallel with increasing relative growth rate (RGR). Fractionation of N- and phosphorus (P) containing compounds indicated that differences in protein N accounted for the treatment differences in total seedling N. Nucleic acid P increased and inorganic P decreased during growth periods, although total seedling P remained constant. Among treated seedlings, height growth was greatest in shaded seedlings: this was probably a result of apical dominance effects because RGR was higher in unshaded seedlings than in shaded seedlings. Thus, the shade treatment alleviated chilling dependent photoinhibition and maximized growth during winter, but limited growth during warmer periods and therefore overall growth. PMID- 12566258 TI - Variability of stem and branch maintenance respiration in a Pinus pinaster tree. AB - The relationship between maintenance respiration (Rm) of woody organs and their structural characteristics was explored in adult Pinus pinaster Ait. trees. We measured Rm on 75 stem and branch segments of different ages (from 3 to 24 years) and diameters (from 1 to 35 cm). The temperature response of Rm was derived from field measurements based on a classical exponential function with Q10 = 2.13. Relationships between Rm and the dimensions of the woody organs were analyzed under controlled conditions in the laboratory. The surface area of a woody organ was a better predictor of Rm than volume, but surface area failed to account for the observed within-tree variability of Rm among stems, branches and twigs. Two simple models were proposed to predict the variability of Rm at 15 degrees C in an adult tree. Model 1, a linear function model based on the dry mass and nitrogen concentration of sapwood and phloem tissues, explained most of the variability of Rm in branches and stems (R2 = 0.97). We concluded that the respective contributions of the phloem and sapwood depend on the location and diameter of the woody organ. Model 2, a power-law function model based on the length, diameter and age of the sample, explained the same variance of Rm as Model 1 and is appropriate for scaling Rm to the stand level. Models 1 and 2 appear to explain a larger variability of Rm than models based on stem area or sapwood mass. PMID- 12566259 TI - Reliance on stored water increases with tree size in three species in the Pacific Northwest. AB - In tall old forests, limitations to water transport may limit maximum tree height and reduce photosynthesis and carbon sequestration. We evaluated the degree to which tall trees could potentially compensate for hydraulic limitations to water transport by increased use of water stored in xylem. Using sap flux measurements in three tree species of the Pacific Northwest, we showed that reliance on stored water increases with tree size and estimated that use of stored water increases photosynthesis. For Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), water stored in xylem accounted for 20 to 25% of total daily water use in 60-m trees, whereas stored water comprised 7% of daily water use in 15-m trees. For Oregon white oak (Quercus garryana Dougl. ex Hook.), water stored in xylem accounted for 10 to 23% of total daily water use in 25-m trees, whereas stored water comprised 9 to 13% of daily water use in 10-m trees. For ponderosa pine (Pinus ponderosa Dougl. ex Laws.), water stored in xylem accounted for 4 to 20% of total daily water use in 36-m trees, whereas stored water comprised 2 to 4% of daily water use in 12-m trees. In 60-m Douglas-fir trees, we estimated that use of stored water supported 18% more photosynthesis on a daily basis than would occur if no stored water were used, whereas 15-m Douglas-fir trees gained 10% greater daily photosynthesis from use of stored water. We conclude that water storage plays a significant role in the water and carbon economy of tall trees and old forests. PMID- 12566260 TI - Intra- and inter-provenance variability in phloem phenols of Picea abies and relationship to a bark beetle-associated fungus. AB - One hundred Norway spruce (Picea abies (L.) Karst.) clones (three ramets per clone) were analyzed for phloem phenol composition and concentration before and 10 days after wound inoculation with sterile malt agar. Fifty clones (Experiment 1) belonged to the same provenance, whereas the remaining clones (Experiment 2) belonged to five provenances from three geographic areas. In Experiment 2, two additional ramets from the same clones were mass inoculated with Ceratocystis polonica (Siem.) C. Moreau to quantify the resistance of each clone. Tree response to wound inoculations was characterized by increased catechin concentration in both experiments, accompanied by increases in astringin and decreases in piceid in Experiment 1. In both experiments, we observed a diverse group of phenolic compounds whose concentrations increased (catechin, astringin) or did not vary (taxifolin glucoside) in response to wound inoculations, whereas concentrations of a homogeneous group of stilbene compounds decreased (piceid) or did not vary (isorhapontin, unidentified stilbene). In Experiment 2, provenances from the alpine and Hercynian-Carpatic areas differed from provenances from the Baltic area with respect to the relative importance of these two groups of compounds, further indicating that the two groupings of phenolic compounds structure the Norway spruce populations. Eighty days after mass inoculation, the percentage of healthy sapwood, which was taken as a measure of tree resistance, indicated that clones from the Baltic area were less resistant to mass inoculations than clones from the alpine and Hercynian-Carpatic areas. We conclude that the degree of resistance of Norway spruce trees to mass inoculations with a bark beetle-associated fungus can be predicted based on the diversity of constitutive phloem phenols and the ability to induce phenol synthesis in response to wounding. PMID- 12566261 TI - Vitality of tree fine roots: reevaluation of the tetrazolium test. AB - Reduction of triphenyltetrazolium chloride (TTC) by tissue to the red-colored insoluble triphenylformazan (TF) is directly linked to the activity of the mitochondrial respiratory chain. Thus, only living tissues should reduce TTC to TF. However, TF production can be detected when the TTC test is applied to control tissues boiled in water. This artifact is mainly the result of the hot ethanol extraction step, which is used to disintegrate the cells and solubilize the TF. We observed that cell wall materials such as cellulose and pectin interact with TTC at temperatures above 60 degrees C, reducing the TTC to TF. By replacing the hot ethanol extraction step with an extraction procedure that involved grinding the boiled tissues and extracting the TF with ethanol at room temperature, the formation of TF was almost entirely eliminated. Application of the modified TTC assay to fine roots of Norway spruce from forest topsoil indicated that the extent of TTC reduction was related to root morphological class as: white fine roots > brown fine roots > black fine roots > boiled fine roots, corresponding to formation of 10.8, 6.1, 0.2 and 0.1 mM TFg(-1)DW, respectively. No significant differences in TTC reduction were recorded between fresh and frozen tissues (frozen in liquid N2) for any fine root class. Application of the modified TTC assay to seedling roots stressed either by drying or by exposure to the toxic metals Cd or Al resulted in significant decreases in TF formation that were related to both the duration of stress and the concentrations of toxic substances, indicating that the modified TTC assay can be used to assess the physiological condition of roots. PMID- 12566262 TI - Tolerance of citrus rootstock seedlings to saline stress based on their ability to regulate ion uptake and transport. AB - Forty-five-day-old seedlings of sour orange (Citrus aurantium L.) and Citrus macrophylla Wester, the most commonly used rootstocks in lemon orchards, were grown in nutrient solutions containing 1 (control), 10, 20, 30 or 60 mM NaCl for 14 days. The effects of salinity on growth, uptake, transport and accumulation of Cl- and Na+ ions in leaves, stem and four root segments were studied. The 60 mM NaCl treatment reduced leaf dry mass more in C. macrophylla (40%) than in sour orange (20%), whereas it reduced root dry mass more in sour orange (36%) than in C. macrophylla (20%). In C. macrophylla, Cl- and Na+ uptake rates were high at the beginning of the saline treatments, but low at the end of the 14-day experiment. In contrast, sour orange showed high uptake rates at the beginning and end of the experiment. In response to increasing salinity, root and shoot concentrations of Cl- and Na+ increased in sour orange, but not in C. macrophylla. Different loading characteristics of Cl- and Na+ were observed between young and old segments of the root system. In general, old root segments reached quasi-steady-states later than young root segments. These results suggest that sour orange and C. macrophylla have different regulatory mechanisms for uptake and transport of Cl- and Na+. PMID- 12566263 TI - Field measurements of root respiration indicate little to no seasonal temperature acclimation for sugar maple and red pine. AB - Increasing global temperatures could potentially cause large increases in root respiration and associated soil CO2 efflux. However, if root respiration acclimates to higher temperatures, increases in soil CO2 efflux from this source would be much less. Throughout the snow-free season, we measured fine root respiration in the field at ambient soil temperature in a sugar maple (Acer saccharum Marsh.) forest and a red pine (Pinus resinosa Ait.) plantation in Michigan. The objectives were to determine effects of soil temperature, soil water availability and experimental N additions on root respiration rates, and to test for temperature acclimation in response to seasonal changes in soil temperature. Soil temperature and soil water availability were important predictors of root respiration and together explained 76% of the variation in root respiration rates in the red pine plantation and 71% of the variation in the sugar maple forest. Root N concentration explained an additional 6% of the variation in the sugar maple trees. Experimental N additions did not affect root respiration rates at either site. From April to November, root respiration rates measured in the field increased exponentially with increasing soil temperature. For sugar maple, long-term Q10 values calculated from the field data were slightly, but not significantly, less than short-term Q10 values determined for instantaneous temperature series conducted in the laboratory (2.4 versus 2.62.7). For red pine, long-term and short-term Q10 values were similar (3.0 versus 3.0). Sugar maple root respiration rates at constant reference temperatures of 6, 18 and 24 degrees C were measured in the laboratory at various times during the year when field soil temperatures varied from 0.4 to 16.8 degrees C. No relationship existed between ambient soil temperature just before sampling and root respiration rates at 6 and 18 degrees C (P = 0.37 and 0.86, respectively), and only a very weak relationship was found between ambient soil temperature and root respiration at 24 degrees C (P = 0.08, slope = 0.09). We conclude that root respiration in these species undergoes little, if any, acclimation to seasonal changes in soil temperature. PMID- 12566264 TI - Effects of NO3- availability on NO3- use in seedlings of three woody shrub species. AB - We determined the effects of short-term cultivation with various amounts of available nitrate nitrogen (NO3-) on NO3- use by woody shrub species. Nitrate concentration ([NO3-]) and nitrate reductase activity (NRA) were measured in leaves and roots of seedlings of Hydrangea hirta (Thunb.) Siebold, Lindera triloba (Sieb. et Zucc.) Blume and Pieris japonica (Thunb.) D. Don. Root [NO3-] increased with increasing NO3- supply in all species, whereas leaf [NO3-] remained low. There were significant correlations between [NO3-] in roots and leaves in all species, but no correlation was found between root NRA and leaf NRA. The low proportion of leaf NO3- assimilation to total NO3- assimilation in all species can be ascribed to the lack of NO3- transport from roots to leaves. In all species, root NRA increased with increasing NO3- supply until reaching a plateau. Species ranking based on maximum root NRA was H. hirta > L. triloba > P. japonica. Root NRA in P. japonica was low, even though root [NO3-] increased with NO3- supply, indicating that NO3- was not an effective N source for this species. The ranking also suggested that H. hirta depended more on NO3- as an N source than L. triloba. The increase in root NRA with increasing NO3- supply was greater in H. hirta than in L. triloba, possibly indicating that a change in NO3- availability has a stronger influence on NO3- use in H. hirta than in L. triloba. PMID- 12566265 TI - Annual pattern of photosynthesis in Scots pine in the boreal zone. AB - To detect seasonal changes in photosynthetic rate in the field, a set of 18,000 photosynthetic measurements made between April and October on three shoots of Scots pine growing near the northern timberline was studied. The measurements were analyzed in the framework of an optimal stomatal control model of photosynthesis, in which irradiance (photosynthetically active radiation, I), air humidity and ambient temperature are driving variables. All driving variables were monitored concomitantly with gas exchange measurements throughout the growing season. The model has nine parameters, of which six were assumed to be constant over the growing season and were fixed based on previous information. The three variable parameters were the initial slope (alpha) and saturation value (gamma) of the light-response curve of carboxylation efficiency in the intercellular cavity, and the cost of transpiration (lambda), in carbon units, regulating the degree of stomatal opening. These parameters could not be estimated independently, nor could their values be satisfactorily found by standard nonlinear regression techniques. A Monte Carlo based simulation procedure was devised to analyze the best-fit parameters and their mutual correlations near the minimum of the residual sum of squares. This was accomplished by replacing the saturation value of the light-response curve with a linearity parameter that determined the shape of the curve. In the best fit solutions, only alpha and lambda varied from day to day, whereas the shape of the curve was constant (i.e., gamma was proportional to alpha). Both alpha and lambda showed consistent patterns from spring to autumn, but the seasonal variation was considerably greater for alpha than for lambda. The optimal stomatal control model with the seven fixed and two daily parameter values gave a good overall fit for photosynthetic rate over the season (PEV > 95%). PMID- 12566266 TI - Growth and nutrient uptake of ectomycorrhizal Pinus sylvestris seedlings in a natural substrate treated with elevated Al concentrations. AB - Models of the effects of elevated concentrations of aluminum (Al) on growth and nutrient uptake of forest trees frequently ignore the effects of mycorrhizal fungi. In this study, we present novel data indicating that ectomycorrhizal mycelia may prevent leaching of base cations and Al. Mycorrhizal and non mycorrhizal Pinus sylvestris L. seedlings were grown in sand obtained from the B horizon of a local forest. In Experiment 1, non-mycorrhizal seedlings and seedlings inoculated with Hebeloma cf. longicaudum (Pers.: Fr.) Kumm. ss. Lange or Laccaria bicolor (Maire) Orton were provided with nutrient solution containing 2.5 mM Al. Aluminum did not affect growth of non-mycorrhizal seedlings or seedlings inoculated with L. bicolor. Seedlings colonized by H. cf. longicaudum had the highest biomass production of all seedlings grown without added Al, but the fungus did not tolerate Al. Shoots of seedlings colonized by L. bicolor had the lowest nitrogen (N) concentrations but the highest phosphorus (P) concentrations of all seedlings. The treatments had small but significant effects on shoot and root Al concentrations. In Experiment 2, inoculation with L. bicolor was factorially combined with the addition of a complete nutrient solution, or a solution lacking the base cations K, Ca and Mg, and solutions containing 0 or 0.74 mM Al. Seedling growth decreased in response to 0.74 mM Al, but the effect was significant only for non-mycorrhizal seedlings. Mycorrhizal seedlings generally had higher P concentrations than non-mycorrhizal seedlings. Aluminum reduced P uptake in non-mycorrhizal plants but had no effect on P uptake in mycorrhizal plants. Mycorrhizal colonization increased the pH of the soil solution by about 0.5 units and addition of Al decreased the pH by the same amount. We conclude that the presence of ectomycorrhizal mycelia decreased leaching of base cations and Al from the soil. PMID- 12566267 TI - Identification and characterization of water-stress-responsive genes in hydroponically grown maritime pine (Pinus pinaster) seedlings. AB - Growth, development and productivity of long-lived organisms such as forest trees are continuously challenged by abiotic stresses, and may also be greatly affected by predicted climatic change. As a first step toward creating stress-resistant maritime pine (Pinus pinaster Ait.) varieties by marker-assisted breeding, we describe the identification and characterization of water-stress-responsive genes in hydroponically grown seedlings that were well watered (-0.08 MPa) or subjected to water deprivation (-0.45 MPa) by the addition of polyethylene glycol. The cDNA amplified fragment-length polymorphism (cDNA-AFLP) technique was used to identify genes regulated by water deprivation. Approximately 4000 transcript-derived fragments (TDFs) were screened, of which 28 increased and 20 decreased in seedlings subjected to water deprivation. Of these 48 TDFs, 62.6% corresponded to proteins of known function, which indicate the main mechanisms involved in the osmotic stress response (photosynthesis, carbohydrate metabolism, cell wall synthesis and plant defense). We found that 16.6% of the 48 TDFs were similar to Arabidopsis thaliana (L.) Heynh gene products, 10.4% were similar to Pinus taeda L. expressed sequence tags (ESTs) and 10.4% did not match any sequences in the public databases. The relative abundance of these transcripts was quantitatively analyzed by reverse northern of both needle and root tissues, confirming the effectiveness of the cDNA-AFLP technique in detecting differentially expressed genes. The identification and characterization of water-stress-responsive genes provide new insights into the nature of the machinery involved in the response to water deprivation in a forest tree. PMID- 12566268 TI - Adaptive significance of C partitioning and regulation of specific leaf area in Betula pendula. AB - Carbon allocation and regulation of specific leaf area (sigma) define key processes underlying the adaptation of plants to varying habitats. In this study, the general principles governing adaptation and a dynamic optimality model of plant adaptation are reviewed. The central new elements of this model are: (i) differential root carbon costs for maintaining a defined nutrient status; (ii) a simple formula for optimal sigma at steady-state as a function of nitrogen (N) status and irradiance; and (iii) generic rules for the time propagation of adapting traits. The model was applied to a large data set compiled by Ingestad et al. (1995) and McDonald et al. (1986a, 1986b) for birch seedlings (Betula pendula Roth) during stationary logarithmic growth and during transient changes in response to a range of irradiances and nutrient supply rates. In the stationary case, large variations in the fraction of leaf dry mass to total dry mass (f(L)), sigma and N concentration were simulated with high accuracy. The independently calibrated model described the temporal response of seedlings following a sharp decrease in N supply, which includes phenomena such as the temporary C accumulation in leaves and damped oscillations in N concentration. Dynamics in sigma were more sensitive to variation in light than in N supply. Nevertheless, adaptive adjustments in f(L), sigma and N concentration were strongly coupled, underlining the relevance of a whole-plant perspective when modeling plant growth and regulation. The high coincidence between model calculations and measured values supports the notion that plant acclimation can be both understood and predicted as a growth-optimizing mechanism. PMID- 12566269 TI - Inducible anatomical defense responses in Norway spruce stems and their possible role in induced resistance. AB - Norway spruce (Picea abies (L.) Karst.) trees were preinoculated with a sublethal dose of the blue-stain fungus Ceratocystis polonica Siem. (C. Moreau) 1 to 52 weeks before they were mass inoculated with the same fungus. Trees pretreated 1 week before mass inoculation had similar, severe symptoms of fungal infection as the control trees. Pretreatment 3, 6 or 9 weeks before mass inoculation resulted in effective protection of the trees, reducing pathogenic symptoms by 63-90% relative to the control trees, whereas pretreatment 52 weeks before mass inoculation gave intermediate protection (44-71% reduction in symptoms). Thus, pretreatment induced resistance to the blue-stain fungus in Norway spruce by a process that requires more than 1 week to become activated and protects trees for at least one year after pretreatment. Pretreatment induced formation of traumatic resin ducts (TDs) in the sapwood and swelling and proliferation of polyphenolic parenchyma cells (PP cells) in the phloem. Trees pretreated 3-9 weeks before mass inoculation had more TDs and showed greater swelling of existing PP cells than control trees or trees pretreated 1 week before mass inoculation. We conclude that induced disease resistance in Norway spruce is probably associated with PP cell activation and TD induction, because resistance was enhanced within the same time frame as the induction of these defense responses. PMID- 12566270 TI - Fruit set dependence on carbohydrate availability in citrus trees. AB - We generated source-sink imbalances by defoliation and sucrose supplementation by stem injection, to investigate effects of carbohydrate availability on fruitlet growth and abscission in cv. Okitsu of Satsuma mandarins (Citrus unshiu (Mak.) Marc.). Partial defoliation promoted fruitlet abscission, whereas sucrose supplementation increased citrus fruit set by more than 10%. Moreover, when applied together, sucrose supplementation counteracted the effect of partial defoliation on fruit set. When sucrose was supplied continuously from flowering until harvest, it increased the concentrations of soluble and insoluble sugars in fruits. We conclude that fruit set in citrus is highly dependent on carbohydrate availability. PMID- 12566271 TI - Reinvigoration of Pinus radiata is associated with partial recovery of juvenile like polyamine concentrations. AB - Polyamine concentrations in consecutive radiata pine (Pinus radiata D. Don) grafts of 30-year-old trees on 1-year-old seedling rootstocks were analyzed by high performance liquid chromatography coupled with fluorescence detection to determine whether reinvigoration is associated with the recovery of polyamine concentrations typical of those in juvenile tissues. Reinvigoration of radiata pine was correlated with the attainment of some, but not all, polyamines characteristic of juvenile trees. In response to reinvigoration, free putrescine, the ratio of free polyamines to low molecular weight polyamine conjugates, and the relative content of putrescine versus spermidine plus spermine increased to values approaching those characteristic of juvenile tissue. In contrast, there was no noticeable change in the concentrations of low molecular weight polyamine conjugates during reinvigoration even though these conjugates increased greatly during tree maturation. We conclude that certain polyamines defined as reinvigoration markers can be used in forestry upgrading programs to assess the morphogenic ability of reinvigorated trees. PMID- 12566272 TI - Early selection improves clonal performance and reduces intraclonal variation of Norway spruce plants propagated by somatic embryogenesis. AB - Height growth during the first and second growth periods (i.e., the June September period in consecutive years) and intraclonal variation were assessed in 13 Norway spruce (Picea abies (L.) Karst.) clones propagated by somatic embryogenesis. The plants were acclimatized and grown in a greenhouse until mid July and then transferred outdoors. The clonal mean heights after the first and second growth periods were lower for somatic embryo plants than for seedlings from corresponding families sown at the time of somatic embryo plant ex vitro transfer, because a large proportion of somatic embryo plants were small. We determined whether certain selection criteria at ex vitro transfer can be used to identify somatic embryo plants with height growth characteristics comparable with those of seedlings. Epicotyl length and presence of lateral roots proved to be important parameters for selection, whereas main root length was less useful. A combined selection for somatic embryo plants with lateral roots and with an epicotyl length exceeding 8 mm resulted in taller plants and reduced intraclonal variation after the first and second growth periods. The growth of somatic embryo plants selected in this way was similar to that of seedlings from the corresponding families. We conclude that selection according to these criteria at ex vitro transfer can result in improved performance of clonal stock propagated by somatic embryogenesis. PMID- 12566273 TI - Severe infantile epilepsies: molecular genetics challenge clinical classification. PMID- 12566274 TI - Axonal damage: a key predictor of outcome in human CNS diseases. AB - Axonal damage has recently been recognized to be a key predictor of outcome in a number of diverse human CNS diseases, including head and spinal cord trauma, metabolic encephalopathies, multiple sclerosis and other white-matter diseases (acute haemorrhagic leucoencephalitis, leucodystrophies and central pontine myelinolysis), infections [malaria, acquired immunodeficiency syndrome (AIDS) and infection with human lymphotropic virus type 1 (HTLV-I) causing HTLV-I-associated myelopathy (HAM)/tropical spastic paraparesis (TSP)] and subcortical ischaemic damage. The evidence for axonal damage and, where available, its correlation with neurological outcome in each of these conditions is reviewed. We consider the possible pathogenetic mechanisms involved and how increasing understanding of these may lead to more effective therapeutic or preventive interventions. PMID- 12566275 TI - Mutations of sodium channel alpha subunit type 1 (SCN1A) in intractable childhood epilepsies with frequent generalized tonic-clonic seizures. AB - A group of infant onset epilepsies manifest very frequent generalized tonic clonic seizures (GTC) intractable to medical therapy, which may or may not be accompanied by minor seizures such as myoclonic seizures, absences and partial seizures. They include severe myoclonic epilepsy in infancy (SMEI) and intractable childhood epilepsy with GTC (ICEGTC). They are commonly associated with fever-sensitivity, family history of seizure disorders and developmental decline after seizure onset. Mutations of the neuronal voltage-gated sodium channel alpha subunit type 1 gene (SCN1A) were recently reported in SMEI patients. To clarify the genotypic differences in this group of epilepsies, we searched for SCN1A abnormalities in 25 patients with SMEI and 10 with ICEGTC, together with the family members of 15 patients. Frameshift mutations in SCN1A were observed in four patients, nonsense mutations in five patients, missense mutations in 21 patients, other mutations in two patients and no mutation in five patients. SMEI patients showed nonsense mutations, frameshifts, or missense mutations, while ICEGTC patients showed only missense mutations. Study of both parents of 11 patients revealed that the mutations in these patients were de novo. However, two mothers had the same missense mutations as their ICEGTC children, and they had generalized epilepsy with febrile seizures plus. Here we suggest that SMEI and ICEGTC represent a continuum with minor phenotypic and genotypic differences. PMID- 12566276 TI - Electrophysiology of focal clonic seizures in humans: a study using subdural and depth electrodes. AB - Focal clonic seizures are a frequent epileptic phenomenon. However, there are few data about their pathogenesis. Eleven patients with focal epilepsy who experienced focal clonic seizures during prolonged video-EEG monitoring were included in this study. Nine patients had subdural electrodes on the precentral gyrus and one patient had additional bilateral subthalamic nucleus (STN) depth electrodes. In five patients, the EEG was co-registered with the EMG of muscles which were involved in the clonic seizures. The frequency, pattern and evolution of the ictal EEG were analysed and their relationship to STN and EMG activity was studied. Focal clonic seizures were always associated with a polyspike-wave pattern in the EEG of the primary motor area (frequency range 1.6-3.4 Hz), while neighbouring electrodes not overlying the precentral gyrus showed different EEG patterns. At seizure onset, the ictal EEG derived from the precentral gyrus consisted of repetitive spiking for 8-28 s (median 19.5 s), accompanied by a continuous increase in muscle tone. This evolved to a pattern of polyspike-wave complexes which were associated with clinical clonus and lasted for 14-202 s (median 30.5 s). The clonic muscle contractions consisted of bursts of compound muscle action potentials (CMAPs) which occurred synchronously in agonistic and antagonistic muscles and were separated by periods of complete muscle relaxation. Each series of CMAPs followed the polyspikes in the EEG with a latency of 17-50 ms. The periods of muscle relaxation occurred during the EEG slow waves. Only some of the cortical spikes were followed by ipsilateral STN spikes. CMAPs followed the cortical polyspikes independently of whether or not STN spikes were seen. The study suggests that focal clonic seizures are focal tonic-clonic seizures. The epileptic clonus consisted of simultaneous contractions of agonistic and antagonistic muscles at regular intervals and was generated by localized polyspike-wave activity in cortical primary motor areas. Activation of the STN did not appear to be an essential component of clonic seizures. PMID- 12566277 TI - Clinical outcomes of hemispherectomy for epilepsy in childhood and adolescence. AB - Hemispherectomy has been performed in the treatment of epilepsy in association with hemiplegia for over 50 years. However, the optimal timing of surgery with respect to age at presentation and the influence of underlying pathology on outcome is only slowly emerging. This study reports on the clinical course and outcomes of 33 children who underwent hemispherectomy at Great Ormond Street Hospital, London, between 1991 and 1997. Age at surgery was 0.33-17 years (median 4.25) with 1-8 years follow-up (median 3.4). The underlying pathology was developmental in 16 (10 hemimegalencephaly, two polymicrogyria, two focal cortical dysplasia, one diffuse cortical dysplasia and one microdysgenesis), acquired in 11 (six middle cerebral artery infarct, three post encephalitis/trauma, and one each of hemiconvulsion-hemiplegia epilepsy and perinatal ischaemic insult) and progressive in six children (four Rasmussen encephalitis, two Sturge-Weber syndrome). At follow-up, 52% were seizure free, 9% experienced rare seizures, 30% showed >75% reduction in seizures and 9% showed <75% seizure reduction or no improvement. Seizure freedom was highest in those with acquired pathology (82%), followed by those with progressive pathology (50%) and those with developmental pathology (31%). However, seizure freedom, rare seizures or >75% reduction in seizures occurred in 100% of those with progressive pathology, 91% of those with acquired and 88% of those with developmental pathology, indicating a worthwhile seizure outcome in all groups. Hemiplegia remained unchanged following surgery in 22 out of 33 children, improved in five and was worse in six. No significant cognitive deterioration or loss of language occurred, and four children showed significant cognitive improvement. Behavioural improvement was reported in 92% of those who had behaviour problems pre operatively. PMID- 12566278 TI - Pathological C-fibres in patients with a chronic painful condition. AB - Little is known about the contribution of C-afferent fibres to chronic painful conditions in humans. We sought to investigate the role of C-fibres in the pathophysiology of pain and hyperalgesia in erythromelalgia as a model disease for chronic pain. Erythromelalgia is a condition characterized by painful, red and hot extremities, and patients often report tenderness on walking. We made microneurographic recordings from single C-fibres in cutaneous fascicles of the peroneal nerve in patients suffering from this disease. All patients had had a pain attack recently and psychophysical signs of allodynia and punctate hyperalgesia were found. We obtained recordings from a total of 103 C-fibres and found significantly lower conduction velocities and increased activity-dependent slowing of the conduction velocity of afferent C-fibres in the patients compared with healthy controls. Furthermore, several units with biophysical properties of mechano-insensitive fibres were pathological, being spontaneously active or sensitized to mechanical stimuli. Since these fibres also mediate the axon reflex flare, their hyperexcitability might account not only for ongoing pain and tenderness but also for redness and warming in this pain syndrome. The changes in conductive properties found in the C-fibres of these patients could be the first signs of a small-fibre neuropathy. This is the first systematic study of single C fibres in patients and it shows an active contribution of mechano-insensitive fibres to chronic pain. PMID- 12566279 TI - The effect of tactile and visual sensory inputs on phantom limb awareness. AB - Multiple sensory stimuli contribute to the conscious awareness of the body. It is well known that limb amputation can result in abnormal body awareness, but the manner in which the CNS constructs and updates a body schema after injury is largely unknown. The purpose of the present study was to systematically evaluate the effects of sensory inputs on phantom limb awareness (PLA) shortly after unilateral upper extremity amputation. The location, quality and intensity of spontaneous and tactile-evoked phantom sensations and awareness were assessed in 13 amputees who were referred sequentially for their initial post-operative rehabilitation. Subjects were tested in three visual conditions: (i) with their eyes open; (ii) with their eyes closed; and (iii) while they viewed their intact hand in a mirror, which created an illusion of their amputated hand (i.e. mirror visual illusion). The mirror illusion was also used to test the effect of combined visual and movement-related stimuli during active voluntary movement. Spontaneous PLA was reported by 12 of the 13 amputees and was not affected by normal visual inputs. Tactile stimulation of the residual limb or face evoked dual percepts in six amputees; i.e. these amputees perceived these touch stimuli as if they were being applied both to the stimulus site and also to a location on the missing limb. This mislocalization phenomenon was most prevalent in the eyes closed condition. Thus, normal vision can strongly override the phantom component of touch-evoked dual percepts. In eight cases, the visual illusion of the missing limb transiently enhanced the spontaneous conscious awareness of the phantom limb. However, the visual illusion did not change the capacity of a tactile stimulus to induce dual percepts. These findings demonstrate that (i) phantom awareness of an amputated body part is common within the 14 months after traumatic upper extremity amputation, (ii) evoked dual percepts are less common than spontaneous PLA, (iii) visual, tactile and sensorimotor systems contribute to PLA, (iv) subtle changes in congruence of sensory information affects both evoked dual percepts and spontaneous PLA, however, (v) sensorimotor information pertaining to the state of the motor system can strongly influence spontaneous PLA, whereas the visual system can predominantly influence evoked PLA. PMID- 12566280 TI - Mutations in the neurofilament light chain gene (NEFL) cause early onset severe Charcot-Marie-Tooth disease. AB - Neurofilament light chain polypeptide (NEFL) is one of the most abundant cytoskeletal components of the neuron. Mutations in the NEFL gene were recently reported as a cause for autosomal dominant Charcot-Marie-Tooth type 2E (CMT2E) linked to chromosome 8p21. In order to investigate the frequency and phenotypic consequences of NEFL mutations, we screened 323 patients with CMT or related peripheral neuropathies. We detected six disease associated missense mutations and one 3-bp in-frame deletion clustered in functionally defined domains of the NEFL protein. Patients have an early onset and often a severe clinical phenotype. Electrophysiological examination shows moderately to severely slowed nerve conduction velocities. We report the first nerve biopsy of a CMT patient with a de novo missense mutation in NEFL, and found an axonal pathology with axonal regeneration clusters and onion bulb formations. Our findings provide further evidence that the clinical variation observed in CMT depends on the gene mutated and the specific type of mutation, and we also suggest that NEFL mutations need to be considered in the molecular evaluation of patients with sporadic or dominantly inherited CMT. PMID- 12566281 TI - Meningeal cells enhance limited CNS remyelination by transplanted olfactory ensheathing cells. AB - Olfactory ensheathing cells (OECs) are candidate cells for transplant-mediated repair of persistent demyelination in diseases such as multiple sclerosis. If this approach is to make the transition from laboratory to clinic, an important issue is the most suitable composition of the OEC transplant. Isolation of OECs involves concurrent isolation of other cell types, and specific selection techniques are required to produce purified OECs. In this study we address whether the purity of the OEC transplant affects their ability to remyelinate. Surprisingly, we find that a purified preparation of OECs, selected on the basis of low-affinity nerve growth factor receptor (p75) expression, results in less extensive remyelination than an unpurified preparation following transplantation into areas of persistent demyelination in rodent CNS in the X irradiation/ethidium bromide (X-EB) model. A distinctive feature of the unpurified preparation both in vitro and following transplantation is the presence of meningeal cells. When meningeal cells are added to purified OECs there is a significant improvement in the extent of remyelination compared with the purified OECs, although if the cells are present in too great an abundance this beneficial effect is lost. These results highlight the important concept that the regenerative properties of OECs are profoundly influenced by the cells with which they are transplanted. PMID- 12566282 TI - Rostral anterior cingulate cortex dysfunction during error processing in schizophrenia. AB - Previous research has demonstrated that patients with schizophrenia have an impaired ability to monitor erroneous responses to stimuli internally. Event related potential (ERP) studies of error-eliciting tasks indicate that, in healthy adults, the commission of an erroneous response is associated with a fronto-centrally distributed negative voltage component termed the error negativity (Ne) or error-related negativity (ERN). In patients with schizophrenia, the Ne/ERN elicited by errors of commission (EoC) is reduced in amplitude compared with that elicited in healthy participants. Functional MRI (fMRI) studies and source localization analyses of ERP data in healthy participants suggest that EoC are associated with activity in the rostral anterior cingulate cortex (ACC). Using event-related fMRI, we examined the brain activity associated with EoC in a group of 10 patients with schizophrenia and 16 matched healthy participants. Patients were stable, partially remitted, medicated out-patients recruited from the community. Participants performed a Go/NoGo task variant that was shown previously to elicit a reduced Ne/ERN during EoC in patients with schizophrenia relative to healthy participants, as well as robust rostral ACC activation during EoC in healthy participants. Patients with schizophrenia were characterized by relative underactivity in the rostral ACC compared with healthy participants. There was also evidence for more widespread underactivity in the limbic system. In contrast to these regions of relative hypoactivity, patients with schizophrenia demonstrated hyperactivity relative to healthy participants in bilateral parietal cortex during both EoC and correctly rejected NoGo trials. Our results are consistent with previous ERP research demonstrating functional abnormalities during error processing in schizophrenia. In light of the role of the rostral ACC and other limbic structures in mediating affective and motivational behaviour, our results suggest there may be a disturbed affective or motivational response to the commission of errors in schizophrenia. PMID- 12566283 TI - Defective cell cycle control underlies abnormal cortical development in the hydrocephalic Texas rat. AB - There is a significant body of evidence to suggest a physiological role for the CSF in both the developing and adult brain. Our recent studies suggest a critical role for this fluid in the developing brain of the hydrocephalic Texas (H-Tx) rat. A key feature of the foetal-onset hydrocephalus in this rat is obstruction in the flow and/or absorption of fluid that is associated with abnormal development of the cerebral cortex resulting in a reduction in the number of neuronal precursors generated. Cells from the affected cerebral cortex do proliferate in vitro and show dose-dependent responses to growth factor stimulation, suggesting that germinal cells are under inhibitory influences in vivo. We tested the hypothesis that the CSF of the affected brains was responsible for the abnormal development. Cells analysed at the time of extraction from affected brains showed an accumulation of cells in the S-phase of the cell cycle, which was reflected in a concentration of cells containing high levels of DNA in the germinal matrix of histological sections of affected brains. CSF from the lateral ventricle of affected foetal brains not only inhibited in vitro proliferation of normal neuronal progenitors, but it also resulted in an accumulation of cells in the S-phase of the cell cycle mimicking the situation in vivo. Fluid from normal foetal brains did not have this effect. From the work detailed here on the mechanistic basis of the deficient cortical development in the foetal hydrocephalic rat brain, we conclude that the content of the CSF is critical in maintaining germinal matrix function and output and, therefore, that the CSF has a vital role in brain development. PMID- 12566284 TI - Conversion of semantic information into phonological representation: a function in left posterior basal temporal area. AB - A unique feature of Japanese language is that its written sentences consist of both morphograms (kanji) and syllabograms (kana). Despite extensive research by PET, functional MRI and magnetoencephalography, the issues of the difference (or the similarities) between the processing of kanji and kana, and between word reading and object/picture naming have not been resolved as yet. This study investigated the function of the posterior basal temporal area in the language dominant hemisphere in auditory and visual language processing, with special emphasis on semantic and phonological recognition. Subdural electrode grids were placed on the left temporal area of a right-handed woman with intractable temporal lobe epilepsy as part of a pre-surgical evaluation. Her dominant hemisphere for language was shown to be the left on the Wada test. Electric stimulation of 50 Hz was applied to the electrodes during the tasks related to language. Our results showed a clear distinction in the responses and/or performance of the subject depending on the type of characters presented and the tasks employed. Electric stimulation of a localized area in the posterior basal temporal lobe caused neither comprehensive nor productive deficit in the tasks using auditory stimuli. In the tasks using visual stimuli, in contrast, impairments were observed in (i) reading of kanji words and (ii) naming of objects/pictures and geometric designs, but not in (iii) reading of kana, (iv) copying of kanji, kana and geometric designs, and (v) using tools. The subject maintained full comprehension of spoken language, suggesting that the auditory tasks are not processed in the posterior basal temporal area. The fact that the impairment of kanji reading and disturbance of object/picture naming were elicited by electric stimulation of the same area indicates that there is at least one anatomical area that is used commonly for kanji (but not kana) and object processing. The conceptual entity of the test items supposedly was recognized correctly, but the concept failed to be matched to correct phonological representation. The left posterior basal temporal area, therefore, has an important function of connecting visual semantic information into phonological representation. PMID- 12566285 TI - Mutations in the ganglioside-induced differentiation-associated protein-1 (GDAP1) gene in intermediate type autosomal recessive Charcot-Marie-Tooth neuropathy. AB - Mutations in the gene for the ganglioside-induced differentiation-associated protein-1 (GDAP1) on 8q21 recently were reported to cause autosomal recessive Charcot-Marie-Tooth (CMT) sensorimotor neuropathy. Neurophysiology and nerve pathology were heterogeneous in these cases: a subset of GDAP1 mutations was associated with peripheral nerve demyelination, whereas others resulted in axonal degeneration. In this study, we identified two novel mutations disrupting the GDAP1 reading frame. Homozygosity for a single base pair insertion in exon 3 (c.349_350insT) was observed in affected children from a Turkish inbred pedigree. The other novel allele detected in a German patient was a homozygous mutation of the intron 4 donor splice site (c.579 + 1G>A). Patients with GDAP1 mutations displayed severe, early childhood-onset CMT neuropathy with prominent pes equinovarus deformity and impairment of hand muscles. Nerve conduction velocities were between 25 and 35 m/s and peripheral nerve pathology showed axonal as well as demyelinating changes. These findings fitted the definition of intermediate type CMT and further support the view that GDAP1 is vital for both, axonal integrity and Schwann cell properties. PMID- 12566286 TI - Differential remoteness and emotional tone modulate the neural correlates of autobiographical memory. AB - Autobiographical memory relies on complex interactions between episodic memory contents, associated emotions and a sense of self-continuity along the time axis of one's life history. The neural correlates underlying autobiographical memory are known to primarily comprise areas of prefrontal cortex, medial and lateral temporal cortex, as well as posterior cingulate and retrosplenial cortex. By contrast, the effect of encoding and/or storage parameters such as the emotional tone of the memories retrieved or the length of the time-interval between the initial encoding of information and retrieval remains to be clarified. Using blocked design functional MRI and statistical parametric mapping, we investigated the impact of remoteness (factor 1: recent, remote) and emotional valence (factor 2: positive, negative) on the neural correlates of autobiographical memory retrieval. Changes in neural activity (P < 0.05, corrected) related to autobiographical memory retrieval (irrespective of remoteness and emotional tone) relative to baseline were observed bilaterally in medial and lateral temporal, temporal-occipital, posterior cingulate and frontal cortices. Recent (relative to remote) memories were associated with differentially increased neural activity bilaterally in the retrosplenial cortex and the hippocampal region, whereas remote (relative to recent) memories did not show any statistically significant differential neural activations. Positive (relative to negative) memories bilaterally activated the orbitofrontal cortex, the temporal pole, as well as medial temporal areas, with the activation peak being in the entorhinal region. By contrast, negative (relative to positive) memories differentially increased neural activity in the right middle temporal gyrus only. The data suggest differential functional roles for temporal, prefrontal and retrosplenial regions during autobiographical memory retrieval depending on the remoteness and the emotional valence of the memories retrieved. In particular, our findings support the 'classic' model of long-term memory processing, which suggests a time-limited differential involvement of the hippocampus in memory consolidation. Interestingly, the observation of such a time-dependent involvement of the hippocampal region in memory consolidation corresponds to the course of retrograde amnesia observed in demented patients, with the loss of recent memories appearing during early stages of the disease when conspicuous neurofibrillary changes are restricted mainly to the hippocampal and parahippocampal regions. Only during later stages, as the neurofibrillary changes spread out to neocortical association areas, do remote memories also become impaired. We conclude that the brain regions involved in autobiographical memory retrieval are influenced by the triggered memories' emotional significance and their relationship to the individual time axis. PMID- 12566287 TI - The effect of negative emotional context on neural and behavioural responses to oesophageal stimulation. AB - Sensory experience is influenced by emotional context. Although perception of emotion and unpleasant visceral sensation are associated with activation within the insula and dorsal and ventral anterior cingulate gyri (ACG), regions important for attention to and perception of sensory and emotional information, the neural mechanisms underlying the effect of emotional context upon visceral sensation remain unexplored. Using functional MRI, we examined neural responses to phasic, non-painful oesophageal sensation (OS) in eight healthy subjects (seven male; age range 27-36 years) either during neutral or negative emotional contexts produced, respectively, by presentation of neutral or fearful facial expressions. Activation within right insular and bilateral dorsal ACG was significantly greater (P < 0.01) during OS with fearful than with neutral faces. In a second experiment, we measured anxiety, discomfort and neural responses in eight healthy male subjects (age range 22-41 years) to phasic, non-painful OS during presentation of faces depicting either low, moderate or high intensities of fear. Significantly greater (P < 0.01) discomfort, anxiety and activation predominantly within the left dorsal ACG and bilateral anterior insulae occurred with high-intensity compared with low-intensity expressions. Clusters of voxels were also detected in this region, which exhibited a positive correlation between subjective behaviour and blood oxygenation level-dependent effect (P < 0.05). We report the first evidence for a modulation of neural responses, and perceived discomfort during, non-painful visceral stimulation by the intensity of the negative emotional context in which the stimulation occurs, and suggest a mechanism for the effect of negative context on symptoms in functional pain disorders. PMID- 12566288 TI - Gluten ataxia in perspective: epidemiology, genetic susceptibility and clinical characteristics. AB - We previously have described a group of patients with gluten sensitivity presenting with ataxia (gluten ataxia) and suggested that this disease entity may account for a large number of patients with sporadic idiopathic ataxia. We have therefore investigated the prevalence of gluten sensitivity amongst a large cohort of patients with sporadic and familial ataxia and looked at possible genetic predisposition to gluten sensitivity amongst these groups. Two hundred and twenty-four patients with various causes of ataxia from North Trent (59 familial and/or positive testing for spinocerebellar ataxias 1, 2, 3, 6 and 7, and Friedreich's ataxia, 132 sporadic idiopathic and 33 clinically probable cerebellar variant of multiple system atrophy MSA-C) and 44 patients with sporadic idiopathic ataxia from The Institute of Neurology, London, were screened for the presence of antigliadin antibodies. A total of 1200 volunteers were screened as normal controls. The prevalence of antigliadin antibodies in the familial group was eight out of 59 (14%), 54 out of 132 (41%) in the sporadic idiopathic group, five out of 33 (15%) in the MSA-C group and 149 out of 1200 (12%) in the normal controls. The prevalence in the sporadic idiopathic group from London was 14 out of 44 (32%). The difference in prevalence between the idiopathic sporadic groups and the other groups was highly significant (P < 0.0001 and P < 0.003, respectively). The clinical characteristics of 68 patients with gluten ataxia were as follows: the mean age at onset of the ataxia was 48 years (range 14-81 years) with a mean duration of the ataxia of 9.7 years (range 1-40 years). Ocular signs were observed in 84% and dysarthria in 66%. Upper limb ataxia was evident in 75%, lower limb ataxia in 90% and gait ataxia in 100% of patients. Gastrointestinal symptoms were present in only 13%. MRI revealed atrophy of the cerebellum in 79% and white matter hyperintensities in 19%. Forty five percent of patients had neurophysiological evidence of a sensorimotor axonal neuropathy. Gluten-sensitive enteropathy was found in 24%. HLA DQ2 was present in 72% of patients. Gluten ataxia is therefore the single most common cause of sporadic idiopathic ataxia. Antigliadin antibody testing is essential at first presentation of patients with sporadic ataxia. PMID- 12566289 TI - Perception and production of biological movement in patients with early periventricular brain lesions. AB - Recent neuroimaging and psychophysical findings suggest that perception and production of human body motion share a common representational network. In the present study, we address the issue of whether early disorders in production of biological movement correspond to impairment in biological motion perception. By using the simultaneous masking paradigm, we examined visual sensitivity to biological motion in adolescents (aged 13-16 years) who were born very preterm (at 27-33 gestational weeks). In a confidence rating procedure, the presence of a point-light walking figure embedded in a moving mask was judged. The participants differed in their locomotion ability, ranging from normal to a complete walking disability exhibiting signs of leg-dominated bilateral spastic cerebral palsy (BS CP) caused by periventricular leukomalacia (PVL). Irrespective of an ability to produce movement, patients with a similar extent of PVL in the parieto-occipital complex exhibit nearly the same sensitivity to biological motion. Sensitivity correlates negatively with the extent of PVL over the parieto-occipital complex, whereas neither the severity of motor disorder nor the severity of pyramidal tract affection relate significantly to the sensitivity index. The data suggest that perception of biological motion is not substantially affected by an observer's early restrictions in body movement. Instead, the findings favour the assumption that the common network for perception and production of biological motion might be inherent for the brain. Motor experience per se does not appear to be necessary for the visual analysis of human movement. PMID- 12566290 TI - The phenomenology of body image distortions induced by regional anaesthesia. AB - Patients with peripheral nerve or spinal cord lesions frequently report perceptual distortions related to position, shape, texture or temperature of the affected areas. This study aimed to describe the phenomenology of such body image alterations during the course of upper limb, lower limb or spinal anaesthetic blocks in patients (n = 36) undergoing orthopaedic surgery. Multimodal sensory testing and assessment of motor function were performed at regular intervals, and the relationship between the reported body image distortions and the progression of sensory and motor impairment was analysed. We found that perceptual changes concerning the shape and size of the deafferented limb occurred in the great majority of patients. In all of them, illusions of swelling, elongation or shortening of the limb coincided with the impairment of warm, cold and/or pinprick sensations, suggesting that thin myelinated Adelta- and/or unmyelinated C-fibres may provide a source of tonic modulation to the limb's cortical representation. Such perceptual alterations of shape and size of body parts differed clearly from postural illusions in terms of frequency, time course and influence of vision. In addition to perceptual changes in the deafferented area, almost half of the patients felt their unanaesthetized lips and/or mouth swelling during the course of upper limb block, suggesting the unmasking of dynamic interactions between somatotopically adjacent cortical representations. Conflicting sensations could co-exist in the patient's body image, such as the illusion of swelling of a limb, which, at the same time, was felt to be missing. The sense of ownership of the deafferented limb was impaired in some cases. These observations show that the perception of body shape and the awareness of its postural variations are built from different plastic models. They also underline the contribution of peripheral afferent activity to the maintenance of a unified body image. PMID- 12566291 TI - Inhibition of subliminally primed responses is mediated by the caudate and thalamus: evidence from functional MRI and Huntington's disease. AB - Masked prime tasks have shown that sensory information that has not been consciously perceived can nevertheless trigger the preactivation of a motor response. Automatic inhibitory control processes prevent such response tendencies from interfering with behaviour. The present study investigated the possibility that these inhibitory control processes are mediated by a cortico-striatal pallidal-thalamic pathway by using a masked prime task with Huntington's disease patients (Experiment 1) and with healthy volunteers in a functional MRI (fMRI) study (Experiment 2). In the masked prime task, clearly visible left- or right pointing target arrows are preceded by briefly presented and subsequently masked prime arrows. Participants respond quickly with a left or right key-press to each target. Trials are either compatible (prime and target pointing in the same direction) or incompatible (prime and target pointing in different directions). Prior behavioural and electrophysiological results show that automatic inhibition of the initially primed response tendency is reflected in a 'negative compatibility effect' (faster reaction times for incompatible trials than for compatible trials), and is shown to consist of three distinct processes (prime activation, response inhibition and response conflict) occurring within 300 ms. Experiment 1 tested the hypothesis that lesions of the striatum would interrupt automatic inhibitory control by studying early-stage Huntington's disease patients. Findings supported the hypothesis: there was a bimodal distribution for patients, with one-third (choreic) showing disinhibition, manifested as an absent negative compatibility effect, and two-thirds (non-choreic) showing excessive inhibition, manifested as a significantly greater negative compatibility effect than that in controls. Experiment 2 used fMRI and a region of interest (ROI) template-based method to further test the hypothesis that structures of the striatal-pallidal-thalamic pathway mediate one or more of the processes of automatic inhibitory control. Neither prime activation nor response conflict significantly engaged any ROIs, but the response inhibition process led to significant modulation of both the caudate and thalamus. Taken together, these experiments indicate a causal role for the caudate nucleus and thalamus in automatic inhibitory motor control, and the results are consistent with performance of the task requiring both direct and indirect striatal-pallidal thalamic pathways. The finding that Huntington's disease patients with greater chorea were disinhibited is consistent with the theory that chorea arises from selective degeneration of striatal projections to the lateral globus pallidus, while the exaggerated inhibitory effect for patients with little or no chorea may be due to additional degeneration of projections to the medial globus pallidus. PMID- 12566292 TI - Stroke patients have selective muscle weakness in shortened range. AB - Weakness is recognized as a major problem after stroke. This study examined the torque-angle curves of stroke individuals and compared them with those of neurologically normal controls to determine (i) if stroke patients were selectively weak when their muscles were placed in a shortened range and (ii) whether contracture influenced any selective weakness. This descriptive research study measured elbow flexor and extensor torque-angle curves and contracture. Twenty-two stroke subjects who had suffered a stroke 5 months to 6 years ago and 11 neurologically normal controls of similar age participated. Torque-angle curves of the elbow flexors and extensors were determined by measuring maximum isometric torque at 0, 20, 40, 60, 80, 100 and 120 degrees of elbow flexion (0 degrees being full elbow extension), where possible. Contracture of the elbow flexors and extensors was measured as the loss of passive elbow joint range of motion. Repeated measures analysis of variance revealed that the torque-angle curves of stroke subjects (with or without contracture) were significantly different from those of the control subjects for both the elbow flexors (P < 0.05) and extensors (P < 0.05). The stroke subjects appeared relatively weaker when the muscles were in their shortened range. This study confirms that selective weakness exists at short muscle lengths after stroke. The findings of this study help to explain why people after stroke have difficulty functioning when their muscles are in their shortened range. Therefore, strength training should be targeted specifically at muscles at their shortened lengths in order to promote the recovery of function after stroke. PMID- 12566293 TI - Correlation between impaired dexterity and corticospinal tract dysgenesis in congenital hemiplegia. AB - One of the most devastating consequences of early corticospinal lesions is the impaired dexterity that results in a noticeable deficit while manipulating small objects. One purpose of the present study was to investigate the extent to which a deficit in the coordination of fingertip forces when grasping and lifting an object between the thumb and index finger could account for the impaired dexterity in patients with congenital hemiplegia (CH). A second objective was to examine whether, in these patients, deficits in skilled hand movements are correlated with the importance of structural damage to the corticospinal tract. The scaling and coordination of fingertip forces during precision grip was investigated in 16 CH patients (aged 8-19 years) and 16 age- and sex-matched control subjects. Proprioception, stereognosis, pressure sensitivity and motor upper limb function (including digital and manual dexterity) were also assessed quantitatively. The structural damage of the corticospinal tract was estimated by measuring the cross-sectional area of cerebral peduncles with MRI and by calculating an index of symmetry between the two peduncles. In CH patients, a large number of parameters measured during the grip-lift task were significantly different when compared with those found in control subjects. Among those, the duration of the preloading and loading phases was significantly longer in CH patients. In addition, both the dissimilarity and time-shift between the profiles of the grip and load force rates, quantified with the cross-correlation method, were also significantly larger in CH patients; the time-shift was strongly correlated with impaired dexterity. These findings suggest that impaired dextrous finger movements in CH patients may specifically result from their inability to ensure a precise synergy between fingertip forces while manipulating an object. Finally, the finding that the time-shift also correlated with the corticospinal tract dysgenesis, as estimated with the cerebral peduncle asymmetry, argues in favour of a critical role of the corticospinal system in the temporal coordination between different muscles involved in dextrous hand movements. Both digital and manual dexterity were also altered in the non-paretic hand of CH patients. This deficit may reveal the contribution of the lesioned hemisphere to the control of ipsilateral skilled finger movements. PMID- 12566294 TI - Disparity between DNA base excision repair in yeast and mammals: translational implications. AB - One approach to the effective treatment of cancer requires the continued development of novel chemotherapeutic agents to kill tumor cells. Additionally, an element of cancer research has been devoted to understanding DNA repair pathways in hopes of defining the factors that confer resistance to anticancer drugs and developing strategies for modulating repair capacity as a means of overcoming resistance or enhancing sensitivity to cancer treatments. Historically, yeast, particularly Saccharomyces cerevisiae, has been used as a model system for DNA repair analyses. Additionally, it has been used to evaluate drug efficacy and selectivity, and to identify new targets for antitumor drugs. The usefulness of yeast for these types of analyses has been primarily because of it being considered to have well-conserved DNA repair processes among eukaryotes. However, as more information has accumulated in mammalian DNA repair, and particularly in DNA base excision repair (BER), a number of striking differences have emerged between yeast and mammalian (human) repair processes. The BER pathway is essential for the repair of damaged DNA induced by oxidizing and alkylating agents, which are the majority of chemotherapeutic drugs used currently in the clinic. The importance of this pathway in processing DNA damage makes its members potential targets for novel chemotherapeutic agents. However, because the BER process and its main players are remarkably divergent from S. cerevisiae to humans, it is worth keeping these differences in mind if yeast continues to be used as a model or primary system in the screening for potential new human therapeutics. PMID- 12566295 TI - Plasminogen activator inhibitor-2: a molecular biomarker for head and neck cancer progression. AB - Head and neck squamous cell carcinoma (HNSCC) is an aggressive epithelial malignancy in which the early diagnosis of premalignant lesions is known to directly correlate with increased survival. However, only a portion of biopsies showing dysplasia will progress to cancer, and there are no currently accepted criteria for predicting which lesions will progress. Therefore, diagnostic protocols that can identify the lesions that are likely to become HNSCC are required. RNA was isolated from normal keratinocytes, the immortalized but nontumorigenic HaCat cell line, and the tumor cell lines SCC-4, SCC-9, SCC-25, and OSCC-3. The RNA was then labeled and used to probe nylon microarray filters that contained a total of 9184 genes (5295 named and 3889 Expression Sequence Tags). Genes whose expression demonstrated a 3-fold or greater change were considered significant. Comparison of expression profiles from normal, HaCat, and four tumor cell lines revealed changes in gene expression in a total of 508 genes. Of these, 16 genes showed a consistent loss of expression when comparing normal to immortalized keratinocytes. In addition, 10 genes demonstrated a consistent loss of expression in the tumor cell lines only. In this latter group of genes, plasminogen activator inhibitor-2 (PAI-2), a gene whose expression has been linked to cell invasion, was additionally investigated. Altered expression of PAI-2 in the different cultured cells was validated via real-time quantitative PCR. In addition, immunohistochemical evaluation of biopsy samples revealed a high expression of PAI-2 in both normal and dysplastic epithelium with a marked decrease of expression in areas of the biopsies containing HNSCC. These data demonstrate that genomic profiling can then be used to identify potential genotypic/phenotypic biomarkers that may predict which dysplastic lesions are most likely to progress to HNSCC. PMID- 12566296 TI - Loss of BRG1/BRM in human lung cancer cell lines and primary lung cancers: correlation with poor prognosis. AB - A role for the SWI/SNF complex in tumorigenesis based on its requirement for retinoblastoma induced growth arrest and p53-mediated transcription and the appearance of tumors in SWI/SNF-deficient mice. In addition, Western blot data have shown that the SWI/SNF ATPase subunits cell, BRG1 and BRM (BRG1/BRM), are lost in approximately 30% of human non-small lung cancer cell lines. To determine whether loss of expression of these proteins occurs in primary tumors, we examined their expression in 41 primary lung adenocarcinomas and 19 primary lung squamous carcinomas by immunohistochemistry. These analyses showed that 10% of tumors show a concomitant loss of BRG1 and BRM expression. Moreover, patients with BRG1/BRM-negative carcinomas, independent of stage, have a statistically significant decrease in survival compared with patients with BRG1/BRM. This report provides supportive evidence that BRG1 and BRM act as tumor suppressor proteins and implicates a role for their loss in the development of non-small cell lung cancers. PMID- 12566298 TI - Angiotensin I-converting enzyme (ACE) gene polymorphism and breast cancer risk among Chinese women in Singapore. AB - Angiotensin II has been shown to have possible mitogenic and angiogenic effects in human cell lines and animal models of breast cancer. It is converted from its precursor to its active form by the angiotensin I-converting enzyme (ACE). A recent epidemiological study observed lower breast cancer incidence in female users of ACE inhibitors relative to nonusers with comparable cardiovascular conditions. To study the hypothesis that reduced ACE activity is associated with reduced risk of breast cancer, we conducted a nested case-control study within the Singapore Chinese Health Study Cohort to investigate the associations between the ACE A-240T and I/D gene polymorphisms, and breast cancer risk. For this analysis, 189 incident breast cancer cases and 671 female cohort control subjects were compared. The low-activity A and I alleles were the putative "low-risk" alleles. The I/D and A-240T genotypes exhibited significant linkage disequilibrium among Singapore Chinese (contingency coefficient = 0.74; P < 0.001). With adjustment for breast cancer risk factors, women with one or two copies of the low activity A allele showed a statistically significant reduction in risk compared with those with the TT genotype [odds ratio (OR), 0.55; 95% confidence interval (CI), 0.34-0.90]. The risk reduction was enhanced after excluding subjects with medical conditions for which ACE inhibitors are commonly prescribed (OR, 0.49; 95% CI, 0.27-0.89). Comparable results were obtained with respect to the I/D genotype and risk of breast cancer. When the I/D and A-240T genotypes were considered simultaneously, compared with women with the high activity genotypes (either TT or DD or both), those with the low-activity genotypes (presence of both A and I alleles) exhibited lower breast cancer risk (OR, 0.46; 95% CI, 0.27-0.81). Our findings support experimental data implicating ACE and angiotensin II in breast cancer, and suggest that the renin-angiotensin system may serve as a therapeutic target for breast cancer treatment and prevention. PMID- 12566297 TI - Survivin expression in mouse skin prevents papilloma regression and promotes chemical-induced tumor progression. AB - Induction of cutaneous squamous cell carcinoma (SCC) in mice, by topical chemical [9,10-dimethylbenzanthracene (DMBA) and phorbol 12-myristate 13-acetate (PMA)] application, is a multistep process involving papilloma formation and progression to carcinoma. We have generated a transgenic (Tg) mouse [keratin-14 (K14) survivin] with skin expression of survivin, an inhibitor of apoptosis expressed in most human skin cancers and premalignant lesions. K14-survivin mice were resistant to DMBA-induced keratinocyte apoptosis. To investigate the role of survivin and apoptosis in cutaneous carcinogenesis, mice were treated once topically with DMBA followed by twice weekly with PMA for 32 weeks. Surprisingly, tumor formation was less frequent (31% versus 43%) and significantly delayed (P = 0.01) in K14-survivin mice compared with non-Tg littermates. On the other hand, papilloma regression was not observed in Tg mice, whereas 20% of papillomas regressed in non-Tgs; one SCC was generated in Tg mice, whereas none were seen in non-Tgs. To increase tumor formation and SCC in particular, a second experiment was performed with mice on a p53+/- background. Again, DMBA/PMA-induced tumor formation was less (71% versus 89%) and significantly delayed (P = 0.02) in K14 survivin p53+/- animals compared with p53+/- non-Tgs. Papilloma regression was also not observed in Tg p53+/- mice, whereas 10% of papillomas regressed in p53+/ non-Tgs. The rate of papilloma progression to SCC was 21% in Tg p53+/- mice compared with 12% in p53+/- non-Tgs. Papillomas did not reveal significant differences in mitotic or apoptotic indices. Survivin expression was detected in all of the tumors. These results indicate that despite a paradoxical negative effect on tumor formation, survivin expression prevents papilloma regression and promotes conversion to SCC, consistent with its expression in most skin cancers and their precursors. PMID- 12566299 TI - Simultaneous targeting of telomeres and telomerase as a cancer therapeutic approach. AB - Telomeres, which are important for maintaining chromosome integrity and functions, shorten with each cell division. Telomerase, responsible for telomere synthesis, is expressed in approximately 90% of human tumor cells but seldom in normal somatic cells. This study evaluated the hypothesis that simultaneous shortening of telomeres and inhibition of telomerase results in synergistic and tumor-selective cytotoxicity. In telomerase-positive human pharynx FaDu tumor cells, paclitaxel caused telomere erosion (first detected at 1 h) and apoptosis. Expression of antisense to the RNA component of human telomerase (hTR) inhibited telomerase activity, shortened telomere length, reduced cell growth rate, and resulted in a significant higher sensitivity to paclitaxel. Another telomerase inhibitor, 3'-azido-3'-deoxythymidine (AZT), at a concentration that produced little or no cell detachment or apoptosis, inhibited the telomerase activity and enhanced the paclitaxel-induced cell detachment and apoptosis. AZT also enhanced the activity of paclitaxel in mice bearing well-established s.c. FaDu xenograft tumors (i.e., reduced residual tumor size, enhanced apoptotic cell fraction, and prolonged survival time), without enhancing host toxicity. In contrast, AZT did not enhance the paclitaxel activity in the telomerase-negative osteosarcoma Saos 2 cells nor in FaDu cells where telomerase was already suppressed by antisense hTR, confirming that the AZT effect in parent FaDu cells is mediated through telomerase inhibition. These results demonstrate that combined use of agents targeting both telomere and telomerase yielded synergistic activity selective for tumors that depend on telomerase for telomere maintenance. PMID- 12566300 TI - Inhibition of cyclooxygenase-2 by rofecoxib attenuates the growth and metastatic potential of colorectal carcinoma in mice. AB - A large number of epidemiological studies have shown that regular use of aspirinor other nonsteroidal anti-inflammatory drugs (NSAIDs) results in a 40-50% reduced risk of colorectal cancer (CRC). Furthermore, NSAIDs cause the regression of preexisting adenomas in patients with familial adenomatous polyposis and significantly inhibit tumor growth in animal models of CRC. To establish a CRC liver metastasis model, we implanted mouse colon tumor MC-26 cells into the splenic subcapsule of BALB/c mice, after which mice were given either standard chow or chow containing the cyclooxygenase (COX)-2-specific inhibitor rofecoxib, alone or in combination with the standard antineoplastic agents, 5-fluoruracil or irinotecan. After 14 days, mice that were given rofecoxib or irinotecan, but not 5-fluoruracil, had significantly smaller primary tumors and fewer metastases. Rofecoxib, at clinical anti-inflammatory plasma concentrations, enhanced the effects of both antineoplastic agents when used in combination. Biochemical analyses of the primary splenic tumor in rofecoxib-treated mice showed no alteration in COX-1 expression, but significant decreases in the expression of the tumor-promoting proteins COX-2, cyclin D1, cytosolic beta-catenin, matrix metalloproteinases-2 and -9, and vascular endothelial cell- derived growth factor. Rofecoxib also decreased growth-enhancing prostaglandin E(2) and tumor suppressive interleukin-10, whereas antineoplastic interleukin-12 was increased. Two separate survival studies were performed. When mice were fed chow containing 0.01% rofecoxib beginning on day 0 after tumor cell implantation, which achieved clinical anti-inflammatory plasma concentrations, survival time was significantly longer compared with mice given control chow. After 30 days, mortality in the control group was 90%, whereas only one mouse (5%) treated with rofecoxib had died after 30 days. In the second survival study, all of the mice were initially fed with regular chow after tumor cell implantation. On day 7, mice were randomly divided into three dietary groups: control chow, low-dose (0.01%) rofecoxib chow, and high-dose (0.025%) rofecoxib chow. After 28 days, mortality was 100%, 20%, and 10% in control, low-, and high-dose rofecoxib fed groups, respectively. These studies demonstrate that rofecoxib decreases the growth and metastatic potential of CRC in mice through multiple mechanisms. These studies in mice also provide important information that supports the benefit of COX-2 inhibition, not only in the prevention of CRC, but also potentially in the treatment of this common malignancy. Clinical trials will be necessary to assess the utility of COX-2 inhibitors as adjuvant therapy for early-stage disease and as potential agents, either alone or in combination, with more established drugs, for the treatment of refractory CRC. PMID- 12566302 TI - Active immunogene therapy of cancer with vaccine on the basis of chicken homologous matrix metalloproteinase-2. AB - Matrix metalloproteinase (MMP) family, in particular MMP-2, may play a key role in angiogenesis and tumor growth. It is conceivable that the breaking of immune tolerance of MMP-2 should be a useful approach to cancer therapy by active immunity. To test this concept, we constructed a plasmid DNA encoding chicken homologous MMP-2 (c-MMP-2) and control vectors. We found that the vaccine based on chicken homologous MMP-2 as a model antigen could induce both protective and therapeutic antitumor immunity. Autoantibodies against MMP-2 in sera of mice immunized with c-MMP-2 could be found by Western blotting analysis and ELISA assay. There was the deposition of autoantibodies within the tumor. IgG1 and IgG2b were substantially increased in response to c-MMP-2 immunization. The elevation of MMP-2 in the sera of tumor-bearing mice was abrogated with the vaccination of c-MMP-2. Transmigration of human endothelial cells and tumor cells through gelatin-coated filters was inhibited with immunoglobulins isolated from mice immunized with c-MMP-2. The gelatinase activity of MMP-2, including both latent MMP-2 (M(r) 72,000) and active MMP-2 (M(r) 66,000) derived from tumor tissues, was apparently inhibited by the vaccination with c-MMP-2. The antitumor activity and the inhibition of angiogenesis were acquired by the adoptive transfer of the purified immunoglobulins. The antitumor activity and production of autoantibodies against MMP-2 could be abrogated by the depletion of CD4(+) T lymphocytes. Angiogenesis was apparently inhibited within tumor, and chick CAMs angiogenesis was also inhibited. Thus, our findings may provide a vaccine strategy for cancer therapy through the induction of an autoimmune response against MMP-2 in a cross-reaction by the immunization with the single xenogeneic homologous MMP-2 gene and may be of importance in the additional exploration of the application of other xenogeneic homologous genes identified in human and other animal genome projects in cancer therapy. PMID- 12566301 TI - A novel subclass of thalidomide analogue with anti-solid tumor activity in which caspase-dependent apoptosis is associated with altered expression of bcl-2 family proteins. AB - Thalidomide is clinically useful in a number of cancers. Antitumor activity may be related to a number of known properties, including anti-tumor necrosis factor (TNF)-alpha and T-cell costimulatory and antiangiogenic activities. However, it may also involve direct antitumor effects. A series of second generation thalidomide analogues have been separated into two distinct groups of compounds, each with enhanced therapeutic potential, i.e., SelCIDs, which are phosphodiesterase (PDE) type IV inhibitors, and IMiDs, which have unknown mechanism(s) of action. We report here our efforts to determine direct antitumor effects of thalidomide and compounds from both groups. We found that one of the SelCID analogues (SelCID-3) was consistently effective at reducing tumor cell viability in a variety of solid tumor lines but had no effect on non-neoplastic cells. The antitumor activity was independent of known PDE4 inhibitory activity and did not involve cAMP elevation. Growth arrest was preceded by the early induction of G(2)-M cell cycle arrest, which led to caspase 3 mediated apoptosis. This was associated with increased expression of pro-apoptotic proteins and decreased expression of antiapoptotic bcl-2. Furthermore, extensive apoptosis in vivo was detected during SelCID-3-mediated inhibition of tumor growth in a murine xenotransplantation cancer model. Our results suggest that SelCID-3 represents a novel antitumor agent distinct from thalidomide and from previously characterized analogues with therapeutic potential against a range of solid tumors. This effect appears to be mediated via alterations in the expression of bcl-2 family proteins. PMID- 12566303 TI - Imbalancing the DNA base excision repair pathway in the mitochondria; targeting and overexpressing N-methylpurine DNA glycosylase in mitochondria leads to enhanced cell killing. AB - The DNA base excision repair (BER) pathway is responsible for the repair of alkylation and oxidative DNA damage. The short-patch BER pathway, beginning with the simple glycosylase N-methylpurine DNA glycosylase (MPG), is responsible for the removal of damaged bases such as 3-methyladenine and 1,N(6)-ethenoadenine from the DNA after alkylation or oxidative DNA damage. The resulting apurinic site is further processed by the other members in the pathway, resulting in the insertion of the correct nucleotide. If apurinic sites accumulate, they are mutagenic and cytotoxic to the cell. To evaluate its efficacy in sensitizing breast cancer cells to chemotherapy, MPG has been overexpressed in the breast cancer cell line, MDA-MB231. With MPG overexpression, an increase in DNA damage and increased cytotoxicity to methyl methanesulfonate as well as increased apoptosis levels was observed in these cells. Because mitochondrial DNA has been shown to be more sensitive to DNA damage than nuclear DNA, a construct containing mitochondrial-targeted MPG using the human manganese superoxide dismutase mitochondrial-targeting sequence was made. Overexpression of the mitochondrially targeted MPG dramatically increased the breast cancer cells' sensitivity to methyl methanesulfonate. In conclusion, we believe that the increase in sensitivity to DNA damage by overexpression of nuclear MPG is because of an imbalance in the BER pathway, and an even greater increase in cell sensitivity is observed when mitochondrial DNA is targeted. PMID- 12566304 TI - Sulindac sulfide inhibits epidermal growth factor-induced phosphorylation of extracellular-regulated kinase 1/2 and Bad in human colon cancer cells. AB - Colorectal cancer is the second leading cause of cancer death in the United States. Nonsteroidal anti-inflammatory drugs including sulindac are promising chemopreventive agents for colorectal cancer. Sulindac and selective cyclooxygenase (COX)-2 inhibitors cause regression of colonic polyps in familial polyposis patients. Sulindac induces apoptotic cell death in cancer cells in vitro and in vivo. In tumor cells, activation of extracellular-regulated kinase (ERK) 1/2 results in phosphorylation of several ERK1/2 effectors, including the proapoptotic protein Bad. Phosphorylation of Ser112 by ERK1/2 inactivates Bad and protects the tumor cell from apoptosis. Sulindac metabolites and other nonsteroidal anti-inflammatory drugs selectively inhibit ERK1/2 phosphorylation in human colon cancer cells. In this study we show that epidermal growth factor (EGF) strongly induces phosphorylation of ERK1/2 and Bad in HT29 colon cancer cells. EGF-stimulated phosphorylation of ERK and Bad is blocked by pretreatment with U0126, a selective MAP kinase kinase (MKK)1/2 inhibitor. Similarly, pretreatment with sulindac sulfide blocks the ability of EGF to induce ERK1/2 and Bad phosphorylation, but also down-regulates total Bad but not ERK1/2 protein levels. The ability of sulindac to block ERK1/2 signaling by the EGF receptor may account for at least part of its potent growth-inhibitory effects against cancer cells. PMID- 12566305 TI - Rapid induction of apoptosis by combination of flavopiridol and tumor necrosis factor (TNF)-alpha or TNF-related apoptosis-inducing ligand in human cancer cell lines. AB - Flavopiridol is one of the first cyclin-dependent kinase inhibitors undergoing clinical tests. We found that the combination treatment of flavopiridol (100-500 nM) with tumor necrosis factor (TNF)-alpha (10 ng/ml) induced a rapid and eminent apoptosis, 20 +/- 5% in 6-h treatment, in a human non-small cell lung carcinoma cell line, A549, as determined by the increase of sub-G(1) fraction in flow cytometry. A similar observation was also made in human colon cancer cell lines, HCT-116 and HCT-15, but not in Rat2, a rat fibroblast cell line. In A549 cells, the cytotoxic synergy by the combination treatment involved the activation of caspase-1, caspase-3, and caspase-8 and generated huge chromosomal degradation. The treatment schedules were so important that only the treatments of flavopiridol concomitantly with or followed by TNF-alpha showed the pronounced apoptosis in A549 cells. Prior treatment of TNF-alpha inhibited the apoptosis by the following combination treatment, leading to little cell death. Yet, such inhibition was reversed when 100 microM of 5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole, a transcription inhibitor, was present during the TNF-alpha pretreatment, suggesting that the inhibitory pretreatment of TNF-alpha might involve antiapoptotic gene expression at the transcriptional level. TNF-alpha treatment resulted in nuclear factor (NF)-kappa B activation, revealed by NF kappa B activity reporter assay. In contrast, flavopiridol was found to inhibit the NF-kappa B-dependent gene transcription, which might give an explanation for the synergistic effect of flavopiridol with TNF-alpha. TNF-related apoptosis inducing ligand (TRAIL; 100 ng/ml) also caused a rapid and strong cytotoxic synergy with flavopiridol. In contrast to TNF-alpha, however, all of the treatment sequences supported the synergy by TRAIL and flavopiridol. The combination of flavopiridol with TNF-alpha or TRAIL may be of use for the development in cancer therapy. PMID- 12566306 TI - A chimeric humanized single-chain antibody against the type I insulin-like growth factor (IGF) receptor renders breast cancer cells refractory to the mitogenic effects of IGF-I. AB - Insulin-like growth factors (IGFs) stimulate breast cancer proliferation, motility, and survival. The type I IGF receptor (IGF1R) mediates the effects of IGF-I. Thus, inhibition of IGF1R activation could inhibit IGF action in breast cancer cells. A single-chain antibody directed against IGF1R (IGF1R scFv-Fc) has been shown to partially inhibit xenograft growth of MCF-7 cells in athymic mice. In this study, we have examined the effects of scFv-Fc on IGF1R signaling in the estrogen receptor-positive (ER+) MCF-7 breast cancer cells in vitro and in vivo. The antibody stimulated IGF1R activation in vitro in MCF-7 cells and was unable to block IGF-I effects. The antibody also stimulated proliferation of MCF-7 cells in monolayer growth assays. To determine how scFv-Fc could stimulate in vitro growth yet inhibit in vivo tumor growth, we examined the effect of scFv-Fc on IGF1R expression. In MCF-7 cells, scFv-Fc down-regulated IGF1R levels after 2 h, and the levels were greatly reduced after 24 h. In contrast, IGF-I treatment over the same time period did not affect IGF1R levels. Twenty-four-h pretreatment of cells with scFv-Fc blocked IGF-I mediated phosphorylation of insulin receptor substrate-1 and subsequent extracellular signal-regulated kinase 1/extracellular signal-regulated kinase 2 and phosphatidylinositol 3'-kinase activation. In contrast, cells treated with 5 nM IGF-I for 24 h still retained the ability to further activate downstream signaling pathways in response to IGF-I. Moreover, pretreatment of MCF-7 cells with scFv-Fc rendered them refractory to further proliferation induced by additional IGF-I. Twenty-four-h pretreatment of cells with scFv-Fc also inhibited IGF-I stimulated anchorage-independent growth. scFv Fc did not enhance antibody-dependent cell-mediated cytotoxicity. In vivo, treatment of mice bearing MCF-7 xenograft tumors with scFv-Fc resulted in near complete down-regulation of IGF1R. Our data show that scFv-Fc stimulates biochemical activation of IGF1R, then causes receptor down-regulation, making MCF 7 cells refractory to additional IGF-I exposure. These results indicate that such chimeric single-chain antibodies against IGF1R have future potential in breast cancer therapy by causing down-regulation of receptor. PMID- 12566307 TI - Frequent display of human papillomavirus type 16 E6-specific memory t-Helper cells in the healthy population as witness of previous viral encounter. AB - Genital human papillomavirus (HPV) infection is common and the majority of infected individuals successfully deal with this virus. Clearance of HPV is presumably mediated by T cells but HPV-16-specific T-cell memory was usually detected in patients with progressive disease and not in healthy subjects, suggesting that HPV-immunity comes too late. We now show the presence of HPV-16 E6-specific memory T-helper (Th) responses in a major fraction (12 of 20) of healthy individuals by application of the IFN-gamma-ELISPOT assay. Although nearly all E6-peptides were recognized, the majority of the responders targeted peptide sequences of the COOH-terminal half (E6(81-158)) of HPV-16 E6. In a direct comparison, the presence of HPV-16 E6-specific T cells coincided with HPV 16 E2-specific T-cell reactivity in healthy individuals, whereas hardly any HPV 16 E7-specific Th immunity was found. This indicates that the induction of T-cell reactivity against HPV-16 E7 is suboptimal during infection when compared with that against HPV-16 E2 and HPV-16 E6. In conclusion, the presence of HPV-16 E6 specific Th memory in the healthy population demonstrates that HPV infection leads to T-cell immunity against immediate early proteins expressed during infection. Because this HPV-16 E6-specific T-cell immunity was frequently detected in healthy subjects, our data suggest that the observed IFN-gamma producing proliferating T cells circulating in the peripheral blood play a role in protection against persistent HPV infection and associated development of malignancies. PMID- 12566308 TI - Non-small cell lung cancer-derived soluble mediators enhance apoptosis in activated T lymphocytes through an I kappa B kinase-dependent mechanism. AB - T lymphocyte survival is critical for the development and maintenance of an effective host antitumor immune response; however, the tumor environment can negatively impact T-cell survival. Lymphocytes exposed to tumor supernatants (TSNs) were evaluated for apoptosis after mitogen stimulation. TSN was observed to significantly enhance phorbol 12-myristate 13-acetate/ionomycin- and anti-CD3 stimulated lymphocyte apoptosis. Enhanced lymphocyte apoptosis was associated with an impairment of nuclear factor kappa B nuclear translocation and diminished I kappa B alpha degradation. In lymphocytes stimulated after exposure to TSNs, cytoplasmic I kappa B alpha persisted as a result of alterations in I kappa B kinase (IKK) activity. Accordingly, although there were no apparent differences in IKK component concentrations, lymphocytes preexposed to TSNs exhibited markedly reduced IKK activity. We conclude that non-small cell lung cancer derived soluble factors promote apoptosis in activated lymphocytes by an IKK dependent pathway. PMID- 12566309 TI - Genotypic and phenotypic characterization of a putative tumor susceptibility gene, GNMT, in liver cancer. AB - Glycine N-methyltransferase (GNMT), a multifunctional protein involved in the maintenance of the genetic stability, is often down-regulated in hepatocellular carcinoma (HCC). Using genotypic characterization of GNMT in hepatoma cell lines and in a Taiwanese population with a high incidence of liver cancer we have investigated the role of this gene in the progression of liver cancer. Six novel polymorphisms, including two short tandem repeats, one 4-nucleotide insertion/deletion polymorphism, and three single nucleotide polymorphisms, in GNMT were identified in this study. The rates of loss of heterozygosity at the GNMT locus in pairs of normal and tumor tissue from the HCC patients were approximately 36-47%. In addition, the observed heterozygosity of GNMT decreases in tumor adjacent liver DNA from HCC patients compared with that observed in blood DNA from normal individuals and HCC patients. This may result from the early event of loss of heterozygosity within the GNMT gene in the liver tissues of HCC patients. However, in this study, we did not observe the association of polymorphic GNMT alleles as inherited risk factors for HCC. We also elucidated the functional impact of genetic markers in the GNMT promoter by performing luciferase reporter gene and gel mobility shift assays. The results indicate that two polymorphisms, short tandem repeat 1 and insertion/deletion polymorphism, in the promoter region could cause allelic specific effects on the transcriptional activity of GNMT. The risk genotypes of GNMT, which presumably have a lower expression level, as estimated from in vitro functional studies, are over represented in tumor-adjacent tissues from HCC patients. In summary, our results suggest that GNMT alteration may be an early event in HCC development and that GNMT could be a new tumor susceptibility gene for HCC. PMID- 12566310 TI - Computational analysis and experimental validation of tumor-associated alternative RNA splicing in human cancer. AB - A genome-wide computational screen was performed to identify tumor-associated alternative RNA splicing isoforms. A BLAST algorithm was used to compare 11,014 genes from RefSeq with 3,471,822 human expressed sequence tag sequences. The screen identified 26,258 alternative splicing isoforms of which 845 were significantly associated with human cancer, and 54 were specifically associated with liver cancer. Furthermore, canonical GT-AG splice junctions were used significantly less frequently in the alternative splicing isoforms in tumors. Reverse transcription-PCR experiments confirmed association of the alternative splicing isoforms with tumors. These results suggest that alternative splicing may have potential as a diagnostic marker for cancer. PMID- 12566311 TI - Regional delivery and selective expression of a high-activity yeast cytosine deaminase in an intrahepatic colon cancer model. AB - A major potential limitation to the success of enzyme prodrug gene therapy is the toxicity that could result from gene expression in normal tissues. In this study, we investigated the use of an enhanced human carcinoembryonic antigen (CEA) promoter for yeast cytosine deaminase (yCD), which converts 5-fluorocytosine to 5 fluorouracil, to increase targeting while maintaining activity both in cell culture and in nude rats bearing intrahepatic xenografts. We found that an enhanced CEA-yCD adenoviral vector can achieve significantly greater yCD expression in CEA-expressing colon carcinoma cell lines (LoVo, HT29, and CaCo2) compared with a nonspecific Rous sarcoma virus-yCD virus. In contrast, infection with CEA-yCD led to lower or equivalent yCD expression in normal hepatocytes or fibroblasts compared with that produced by the RSV-yCD. Adenovirus administered in the portal vein or the hepatic artery of nude rats bearing intrahepatic LoVo colon carcinomas could mediate beta-galactosidase expression equally in liver and tumors under the control of cytomegalovirus, a nonspecific promoter. However, infusion of CEA-yCD virus markedly increased yCD expression in tumors over normal liver (>4-fold) measured both by levels of mRNA and yCD activity. Moreover, the efficiency of 5-fluorocytosine conversion into 5-fluorouracil in tumors was significantly higher than that in normal liver ( approximately 3-fold) in rats receiving portal venous viral infusion of CEA-yCD and subsequent 5FC treatment. Thus, an enhanced CEA promoter can preferentially stimulate yCD gene expression in CEA-expressing cells in vivo. Such tumor-specific expression should prove useful in colorectal cancer gene therapy to achieve selective prodrug conversion in tumors. PMID- 12566312 TI - Hypomethylation of the synuclein gamma gene CpG island promotes its aberrant expression in breast carcinoma and ovarian carcinoma. AB - Recent studies indicate that synuclein gamma (SNCG) gene, located in chromosome 10, participates in the pathogenesis of the breast and ovary. SNCG, also known as breast cancer-specific gene 1 (BCSG1), is not expressed in normal mammary or ovarian surface epithelial cells but is highly expressed in the vast majority of advanced staged breast and ovarian carcinomas. When overexpressed, SNCG significantly stimulates breast cancer proliferation and metastasis. To fully understand the molecular mechanisms underlying the abnormal expression of SNCG in neoplastic diseases, in this study, we extensively examined the methylation status of a CpG island located in exon 1 of SNCG gene in a panel of breast and ovarian tumor-derived cell lines to determine whether DNA methylation plays a crucial role in SNCG expression. In vivo bisulfite DNA sequencing of genomic DNA isolated from breast cancer cell lines showed that the 15 CpG sites within the CpG island were completely unmethylated in all SNCG-positive cell lines (5 of 5), but were densely and heterogeneously methylated in the majority of SNCG-negative cell lines (3 of 4). The methylation occurred primarily at the CpG sites 2, 5, 7, and 10-15. Similarly, we observed a strong correlation of hypomethylation of the CpG island and SNCG expression in ovarian cancer cell lines (5 of 5). Intriguingly, the methylation pattern in ovarian cancer cells is different from that in breast cancer cells. In SNCG-nonexpressing ovarian cancer cells, all 15 of the CpG sites were completely methylated instead of selective methylation at certain sites shown in breast cancer cells, thereby suggesting a tissue-specific methylation pattern. A correlation between hypomethylation of the exon 1 and expression of SNCG mRNA was also observed in primary breast tumor tissues. The importance of DNA methylation in the control of SNCG expression in cancer cells is further strengthened by demonstration of re-expression of SNCG mRNA in SNCG negative ovarian and breast cancer cells with a demethylating agent 5-aza-2' deoxycytidine. In addition, we demonstrate that inhibition of cell growth leads to a decreased mRNA expression and an increased DNA methylation of SNCG gene. Taken together, these new findings strongly suggest that DNA hypomethylation is a common mechanism underlying the abnormal expression of this candidate oncogene in breast and ovarian carcinomas. PMID- 12566313 TI - Keratinocyte-specific Pten deficiency results in epidermal hyperplasia, accelerated hair follicle morphogenesis and tumor formation. AB - PTEN is a tumor suppressor gene mutated in many human cancers. We used the Cre loxP system to generate a keratinocyte-specific null mutation of Pten in mice (k5Pten(flox/flox) mice). k5Pten(flox/flox) mice exhibit wrinkled skin because of epidermal hyperplasia and hyperkeratosis and ruffled, shaggy, and curly hair. Histological examination revealed that skin morphogenesis is accelerated in k5Pten(flox/flox) mice. Within 3 weeks of birth, 90% of k5Pten(flox/flox) mice die of malnutrition possibly caused by hyperkeratosis of the esophagus. All k5Pten(flox/flox) mice develop spontaneous tumors within 8.5 months of birth, and chemical treatment accelerates the onset of tumors. k5Pten(flox/flox) keratinocytes are hyperproliferative and resistant to apoptosis and show increased activation of the Pten downstream signaling mediators Akt/protein kinase B (PKB) and extracellular signal-regulated kinase. Pten is thus an important regulator of normal development and oncogenesis in the skin. PMID- 12566314 TI - UBP41 is a proapoptotic ubiquitin-specific protease. AB - We have isolated Ubp41, a ubiquitin-specific protease, in a screen for proapoptoticgenes. We found that overexpression of Ubp41 is sufficient to elicit all features of apoptosis in human cells. In contrast, an enzymatically defective UBP41 mutant and homologous ubiquitin-processing protease family members did not significantly induce cell death. Overexpression of Ubp41 resulted in a strong deubiquitination of a broad range of proteins, but surprisingly did not lead to a stabilization of protein substrates known to be regulated by the ubiquitin proteasome system such as the cell cycle factors p21 and p27. Hence, in contrast to the proteasome inhibitor MG132, Ubp41 overexpression did not arrest cells in G(2)/M. Rather, overexpression of hUbp41 seems to interfere with the ubiquitin system and to cause the activation of apoptosis pathways by stabilizing specific substrates. Hence, for the first time we found that a member of the deubiquitinating enzymes has a direct proapoptotic activity additionally tightening the connection between apoptosis and the ubiquitin-proteasome system. PMID- 12566315 TI - A chromosome 3-encoded repressor of the human telomerase reverse transcriptase (hTERT) gene controls the state of hTERT chromatin. AB - Telomerase is crucial for human carcinogenesis. The limiting component of telomerase activity is telomerase reverse transcriptase (hTERT), undetectable in differentiated somatic cells but present in most tumor cells. There is evidence that hTERT transcription is shut down by a repressor in normal cells, but the mechanisms that turn on or maintain expression in tumor cells are not understood. To identify cis-acting regulatory elements, we scanned the hTERT gene for nuclease sensitive sites. In tumor cells and in in vitro transformed fibroblasts that contain hTERT mRNA, we detected a pattern of nuclease-sensitive sites in the second intron different from that in normal fibroblasts. To test whether the chromatin state characterized by the increased nuclease sensitivity plays a role in tumor-specific hTERT expression, we used a telomerase-positive breast carcinoma line, 21NT. Introduction of a normal chromosome 3 into these cells is known to down-regulate hTERT expression, probably through transcriptional silencing. 21NT cells displayed a similar pattern of micrococcal nuclease (MNase) sensitivity to other telomerase-positive lines, whereas the hTERT chromatin of the chromosome 3-hybrids resembled that of normal fibroblasts. In segregants that had lost the normal chromosome 3, the MNase sensitivity pattern characteristic of telomerase-positive cells was restored, and some (but not all) re-expressed the hTERT gene. The simplest model compatible with these results, and with data on the mapping of an hTERT repressor on chromosome 3, is that hTERT expression in tumor cells depends on an open state of intron 2-chromatin. We propose that, during the development of the breast carcinoma from which the 21NT cell line was derived, loss of function of this repressor led to chromatin remodeling necessary (but probably not sufficient) for expression of the hTERT gene. An improved understanding of the precise mechanism of hTERT dysregulation in human cancer may well find applications in the development of antitelomerase cancer therapy. PMID- 12566316 TI - The variant hepatocyte nuclear factor 1 activates the P1 promoter of the human alpha-folate receptor gene in ovarian carcinoma. AB - The alpha folate receptor (alpha FR) is a membrane glycoprotein that binds folates, and mediates their uptake and that of antifolate drugs. alpha FR is absent on ovarian surface epithelium (OSE) but is detectable during early transforming events in this epithelium, with increasing expression levels in association with tumor progression. Analysis of transcriptional regulation of the alpha FR gene have revealed two promoter regions, P1 and P4, flanking exons 1 and 4, respectively, and a requirement for three SP1 sites and an INR element for optimal P4 activity. Here, we focused on the P1 transcription regulation in ovarian carcinoma cells. RNase protection assay indicated that the 5' untranslated region is heterogeneous because of different start sites and alternative splicing of exon 3. A core region of the P1 promoter was sufficient for maximal promoter activity in ovarian carcinoma cell lines but not in OSE cells or in alpha FR-nonexpressing cell lines. Deletion and mutation analysis of this core promoter identified a cis-regulatory element at position +27 to +33 of the untranslated exon 1, which is responsible for maximum P1 activity. This element formed an abundant DNA-protein complex with nuclear proteins from ovarian cancer cells but not from other cell lines or OSE cells. Competition experiments and supershift assays demonstrated binding of the P1 cis-regulatory element by a transcription factor involved in embryonic development, the variant hepatocyte nuclear factor-1 (vHNF1). Analysis of RNA from various cell lines and surgical specimens confirmed that vHNF1 is expressed in ovarian carcinomas. Thus, vHNF1 regulates tissue-specific transcription in ovarian carcinoma. PMID- 12566317 TI - Small unstable apoptotic protein, an apoptosis-associated protein, suppresses proliferation of myeloid cells. AB - Apoptosis, or programmed cell death, is a process where developmental or environmental stimuli activate a genetic program to implement a series of events that culminate in cell death. To study the nature of genes that are induced during the apoptotic death of myeloid precursor cells, representational difference analysis was performed using RNAs derived from 32Dcl3 myeloblastic cells that were proliferating in the presence of IL-3 and cells that were actively undergoing apoptosis as a result of interleukin 3 deprivation for 24 h. This report describes a novel gene [small unstable apoptotic protein (SUAP)] that is up-regulated in these cells after the removal of interleukin 3 and exposure to granulocyte colony stimulating factor. The protein encoded by this gene is a target of the proteasome and does not share homology with other previously characterized proteins. To further define SUAP's role in growth arrest and apoptosis, 32Dcl3 cells that ectopically express SUAP under the control of an inducible promoter were generated and tested for their ability to proliferate under conditions where SUAP expression is induced. These studies show that although the SUAP expressing cells exhibited suppressed proliferation rates, this was not attributable to alterations in cell cycle progression. Rather, SUAP appears to induce the appearance of Annexin V-positive cells, supporting a role for this protein in programmed cell death. PMID- 12566318 TI - Differential in vivo and in vitro expression of vascular endothelial growth factor (VEGF)-C and VEGF-D in tumors and its relationship to lymphatic metastasis in immunocompetent rats. AB - The presence of metastases in regional lymph nodes is a strong indicator of poor patient survival. A number of clinical and experimental studies suggest that tumor-induced lymphangiogenesis driven by vascular endothelial growth factor (VEGF)-C- and/or VEGF-D-induced activation of VEGF receptor (VEGFR)-3 may promote metastasis to regional lymph nodes. Here we show that constitutive VEGF-C and VEGF-D expression by tumor cells of diverse origin grown in tissue culture does not correlate with metastatic potential in vivo. However, tumors derived from cell lines that do not constitutively express VEGF-C or VEGF-D in tissue culture can nevertheless express one or both of these factors. We demonstrate that both tumor and stromal cells can contribute to this expression, suggesting that tumor cell-host interactions determine tumor expression of VEGF-C and VEGF-D. Using immunocompetent rat mammary tumor models, we show in two ways that this expression can promote metastasis via the lymphatics. Firstly, ectopic expression of a soluble VEGFR-3 receptor globulin protein in MT-450 tumor cells that are highly metastatic via the lymphatics blocked VEGF-C and VEGF-D activity and suppressed metastasis formation in both the regional lymph nodes and the lungs. Secondly, ectopic expression in the weakly metastatic NM-081 cell line of a mutant form of VEGF-C that is only able to activate VEGFR-3 strongly promoted metastasis of these cells to the regional lymph nodes and lung. These data show that expression of VEGF-C and VEGF-D in tissue culture does not reflect expression in vivo and that activation of VEGFR-3 in the absence of VEGFR-2 activation is sufficient to promote tumor-induced lymphangiogenesis and metastasis, and they support the notion that blockade of VEGFR-3 activation will be useful as a novel form of cancer therapy. PMID- 12566319 TI - Lanthionine synthetase components C-like 2 increases cellular sensitivity to adriamycin by decreasing the expression of P-glycoprotein through a transcription mediated mechanism. AB - Although the coincidental amplification and accompanying overexpression of bystander genes that neighbor oncogene targets occur frequently during the development of human tumors, little has been done to investigate the functional or biological consequences of amplified bystander gene overexpression. LANCL2 (LANC-like 2) is a bystander gene that is coamplified and overexpressed with epidermal growth factor receptor in approximately 20% of all glioblastomas. This gene has also been designated as Testis Adriamycin Sensitivity Protein because it is most highly expressed in testis and its expression has been noted to increase cellular sensitivity to Adriamycin. Because of the latter association, we have examined potential relationships between LANCL2 and the expression of multidrug resistance (MDR)1, as well as its cognate protein, P-glycoprotein (P-gp), because elevated expression of P-gp is known to increase cell resistance to many cytotoxic drugs, including Adriamycin. Using the Dx5 derivative of MES-SA cells in which P-gp is overexpressed, we show that the level of endogenous P-gp decreases with increased expression of exogenous LanCl-2 and that cells with reduced P-gp show increased sensitivity to Adriamycin. Results from reverse transcription-PCR and MDR1 promoter activity analyses suggest that LanCl-2 transcriptionally suppresses MDR1, and this interpretation of LanCl-2 function is consistent with results from immunofluorescence analysis, which shows that LanCl 2 resides in the nucleus, as well as at the plasma membrane. With respect to this study, our data indicate that LanCl-2 increases cellular sensitivity to Adriamycin by decreasing the expression of P-gp, but more generally, these results indicate that the identification of bystander gene amplification in human tumors can have clinical implications. PMID- 12566320 TI - Regulation of cyclooxygenase-2 expression by the Wnt and ras pathways. AB - Mutations in the adenomatous polyposis coli (APC) gene and K-ras occur in the majority of human colorectal cancers. Loss of functional APC protein activates the Wnt signal transduction pathway, allowing the nuclear accumulation of beta catenin, which then binds to T-cell factor-4 (Tcf-4), causing increased transcriptional activation of downstream target genes. We investigated the hypothesis that the activation of the WNT pathway regulates cyclooxygenase-2 (COX 2). COX-2 was down-regulated after the induction of full-length APC in the HT29 APC cell line. We identified a Tcf-4-binding element (TBE) in the COX-2 promoter that specifically bound to Tcf-4 in an electrophoretic mobility shift assay. COX 2 promoter luciferase activity is down-regulated by APC in a promoter reporter construct containing the, TBE but not with mutant TBE. Mutant beta-catenin expression up-regulated the COX-2 promoter activity and the endogenous COX-2 mRNA expression in HuH7, hepatocellular carcinoma cell line, which is partially abrogated by cotransfection with a dominant-negative Tcf-4 expression vector. Although beta-catenin alone did not increase COX-2 protein to detectable levels in HuH7 cells, coexpression of both mutant beta-catenin and mutant K-ras increased COX-2 protein expression, which is consistent with the previous reports that K-ras can stabilize COX-2 mRNA. Taken together, our data support the hypothesis that COX-2 is down-regulated by APC and up-regulated by nuclear beta catenin accumulation, and additionally implicate the Wnt signal transduction pathway in colon and liver carcinogenesis. PMID- 12566321 TI - We need Romanow's National Drug Agency. PMID- 12566324 TI - Children are not small adults. PMID- 12566325 TI - Deces suite a un implant cochleaire : pas un type b. PMID- 12566326 TI - Meningitis and cochlear implantation. PMID- 12566330 TI - Trends in the place of death of cancer patients, 1992-1997. AB - BACKGROUND: Although many patients with cancer would prefer to die at home, most die in hospital. We carried out a study to describe the yearly trends in the place of death between 1992 and 1997 and to determine predictors of out-of hospital death for adults with cancer in Nova Scotia. METHODS: In this population based study, we linked administrative health data from 2 databases - the Nova Scotia Cancer Centre Oncology Patient Information System and the Queen Elizabeth II Health Sciences Centre Palliative Care Program - for all adults in Nova Scotia who died of cancer from 1992 to 1997. We also used grouped neighbourhood income information from the 1996 Canadian census. Death out of hospital was defined as death in any location other than an acute care hospital facility. We used logistic regression analysis to identify the odds of dying out of hospital over time and to identify factors predictive of out-of-hospital death. RESULTS: A total of 14 037 adults died of cancer during the study period. The data for 101 people were excluded because of missing information regarding place of death. Of the remaining 13 936 people, 10 266 (73.7%) died in hospital and 3670 (26.3%) died out of hospital. Over the study period the proportion of people who died out of hospital rose by 52%, from 19.8% (433/2182) in 1992 to 30.2% (713/2359) in 1997. Predictors associated with out-of-hospital death included year of death (for 1997 v. 1992, adjusted odds ratio [OR] 1.8, 95% confidence interval [CI] 1.5 2.0), female sex (adjusted OR 1.2, 95% CI 1.1-1.3), age (for > or = 85 v. 18-44 years, adjusted OR 2.2, 95% CI 1.7-2.8), length of survival (for 61-120 v. < or =60 days, adjusted OR 2.2, 95% CI 1.8-2.6; for 121-180 v. < or =60 days, adjusted OR 2.5, 95% CI 2.2-2.8), having received palliative radiation (adjusted OR 0.8, 95% CI 0.7-0.9) and region of death (Cape Breton v. Halifax, adjusted OR 0.5, 95% CI 0.5-0.6). Among Halifax residents, registration in the Palliative Care Program was also a significant predictor of out-of-hospital death (adjusted OR 1.4, 95% CI 1.2-1.7). Tumour group, neighbourhood income and residence (urban v. rural) were not predictive of out-of-hospital death in multivariate analysis. INTERPRETATION: Over time, more patients with cancer, especially women, elderly people and people with longer survival after diagnosis, died outside of hospital in Nova Scotia. PMID- 12566329 TI - Geography and service supply do not explain socioeconomic gradients in angiography use after acute myocardial infarction. AB - BACKGROUND: Socioeconomic status appears to be an important predictor of coronary angiography use after acute myocardial infarction. One potential explanation for this is that patients with lower socioeconomic status live in neighbourhoods near nonteaching hospitals that have no catheterization capacity, few specialists and lower volumes of patients with acute myocardial infarction. This study was conducted to determine whether the impact of socioeconomic status on angiography use would be lessened by considering variations in the supply of services. METHODS: We examined payment claims for physician services, hospital discharge abstracts and vital status data for 47 036 patients with acute myocardial infarction admitted to hospitals in Ontario between April 1994 and March 1997. Neighbourhood income of each patient was obtained from Canada's 1996 census. Using multivariate hierarchical logistic regression and adjusting for baseline patient and physician factors, we examined the interaction among hospital and regional characteristics, socioeconomic status and angiography use in the first 90 days after admission to hospital for acute myocardial infarction. RESULTS: Within each hospital and geographic subgroup, crude rates of angiography rose progressively with increases in neighbourhood income. After adjusting for sociodemographic, clinical and physician characteristics, hospitals with on-site angiography capacity (adjusted odds ratio [OR] 1.88, 95% confidence interval [CI] 1.52-2.33), those with university affiliations (adjusted OR 1.60, 95% CI 1.27 2.01) and those closest to tertiary institutions (adjusted OR 1.57, 95% CI 1.32 1.87) were all associated with higher 90-day angiography use after acute myocardial infarction. However, the relative impact of socioeconomic status on 90 day angiography use was similar whether or not hospitals had on-site procedural capacity (interaction term p = 0.68), had university affiliations (interaction term p = 0.99), were near tertiary facilities (interaction term p = 0.67) or were in rural or urban regions (interaction term p = 0.90). INTERPRETATION: Socioeconomic status was as important a predictor of angiography use in hospitals with ready access to cardiac catheterization facilities as it was in those without. The socioeconomic gradient in the use of angiography after acute myocardial infarction cannot be explained by the distribution of specialists or tertiary hospitals. PMID- 12566331 TI - Stress, satisfaction and burnout among Dutch medical specialists. AB - BACKGROUND: Stress and stress-related illnesses are increasing among medical specialists. This threatens the quality of patient care. In this study we investigated (a) levels of job stress and job satisfaction among medical specialists, (b) factors contributing to stress and satisfaction and (c) the effect of stress and satisfaction on burnout. METHODS: A questionnaire was mailed to a random sample of 2400 Dutch medical specialists. Measures included job stress, job satisfaction, burnout, personal characteristics, job characteristics and perceived working conditions. RESULTS: The final response rate was 63%. Of the respondents, 55% acknowledged high levels of stress, and 81% reported high job satisfaction. Personal and job characteristics explained 2%-6% of the variance in job stress and satisfaction. Perceived working conditions were more important, explaining 24% of the variance in job stress and 34% of the variance in job satisfaction. Among perceived working conditions, the interference of work on home life (odds ratio [OR] 1.54, 95% confidence interval [CI] 1.35-1.76) and not being able to live up to one's professional standards (OR 1.57, 95% CI 1.37 1.80) were most related to stress. Feeling poorly managed and resourced (OR 2.07, 95% CI 1.76-2.43) diminished job satisfaction. Burnout was explained by both high stress and low satisfaction (41% of variance explained) rather than by stress alone. INTERPRETATION: Our study showed a protective effect of job satisfaction against the negative consequences of work stress as well as the importance of organizational rather than personal factors in managing both stress and satisfaction. PMID- 12566332 TI - Emergency department overcrowding and ambulance transport delays for patients with chest pain. AB - OBJECTIVE: Emergency department overcrowding sometimes results in diversion of ambulances to other locations. We sought to determine the resulting prehospital delays for cardiac patients. METHODS: Data on consecutive patients with chest pain who were transported to Toronto hospitals by ambulance were obtained for a 4 month period in 1997 and a 4-month period in 1999, which represented periods of low and high emergency department overcrowding respectively. Multivariate analyses were used to model 90th percentile system response (initiation of 9-1-1 call to arrival on scene), on-scene (arrival on scene to departure from scene) and transport (departure from scene to arrival at hospital) intervals. Predictor variables were study period (1997 or 1999), day of the week, time of day, geographic location of the patient, dispatch priority, case severity, return priority and number of other patients with chest pain transported within 2 hours of the index transport. RESULTS: A total of 3609 patients (mean age 66.3 years, 50.3% female) who met the study criteria were transported by ambulance during the 2 study periods. There were no significant differences in patient characteristics between the 2 periods, despite the fact that more patients were transported during the second period (p < 0.001). The 90th percentile system response interval increased by 11.3% from the first to the second period (9.7 v. 10.8 min, p < 0.001), whereas the on-scene interval decreased by 8.2% (28.0 v. 25.7 min, p < 0.001). The longest delay was in the transport interval, which increased by 28.4% from 1997 to 1999 (13.4 v. 17.2 min, p < 0.001). In multivariate analyses, the study period (1997 v. 1999) remained a significant predictor of longer transport interval (p < 0.001) and total prehospital interval (p = 0.004). INTERPRETATION: An increase in overcrowding in emergency departments was associated with a substantial increase in the system response interval and the ambulance transport interval for patients with chest pain. PMID- 12566333 TI - Why a journal oversight committee? PMID- 12566334 TI - Toward effective Canadian public-private partnerships in health research. PMID- 12566335 TI - Current concepts in the diagnosis and management of Parkinson's disease. AB - Parkinson's disease is a progressive neurological disorder characterized by rest tremor, bradykinesia, rigidity and postural instability. The cause is unknown, but growing evidence suggests that it may be due to a combination of environmental and genetic factors. Treatment during the early stage of Parkinson's disease has evolved, and evidence suggests that dopamine agonist monotherapy may prevent the response fluctuations that are associated with disease progression. L-dopa therapy, however, remains the most efficacious treatment. Treatment during the advanced stage focuses on improving control of a number of specific clinical problems. Successful management of motor response fluctuations (e.g., "wearing off," on-off fluctuations, nighttime deterioration, early morning deterioration and dyskinesias) and of psychiatric problems is often possible with specific treatment strategies. Surgical treatment is an option for a defined patient population. PMID- 12566336 TI - Out of the cold: management of hypothermia and frostbite. PMID- 12566337 TI - Drinking and driving. PMID- 12566338 TI - What type of exercise prevents cardiovascular disease in postmenopausal women? PMID- 12566339 TI - Intussusceptions in a man with Peutz-Jeghers syndrome. PMID- 12566341 TI - A very political art. PMID- 12566342 TI - How he had been. PMID- 12566344 TI - Dispute over use of MDs' prescribing information heading to court. PMID- 12566346 TI - Ottawa enjoys broad MD support as Kyoto protocol ratified. PMID- 12566347 TI - Free fall in HIV/AIDS totals continues. PMID- 12566348 TI - Smoking finally on the wane in Nova Scotia? PMID- 12566350 TI - Fight for Ottawa's money will be fierce. PMID- 12566351 TI - After 6 years, vindication for CMA past president. PMID- 12566353 TI - Morgentaler's call for secular-only hospitals earns tepid response. PMID- 12566354 TI - Drawing blood from unconscious accident victims now legal in UK. PMID- 12566355 TI - The staggering cost of illness and injury. PMID- 12566356 TI - New committee to oversee relationship between CMA, CMAJ. PMID- 12566358 TI - Predicting sudden death risk for heart failure patients in the implantable cardioverter-defibrillator age. PMID- 12566359 TI - Paclitaxel coating reduces in-stent intimal hyperplasia in human coronary arteries: a serial volumetric intravascular ultrasound analysis from the Asian Paclitaxel-Eluting Stent Clinical Trial (ASPECT). AB - BACKGROUND: The aim of this study was to use serial volumetric intravascular ultrasound (IVUS) to evaluate the effect of a paclitaxel coating on in-stent intimal hyperplasia (IH). METHODS AND RESULTS: Patients were randomized to placebo (bare metal stents) or 1 of 2 doses of paclitaxel (low dose: 1.28 microg/mm2; high dose: 3.10 microg/mm2). Complete post-stent implantation and follow-up IVUS were available in 81 patients, including 25 control patients and in 28 receiving a low-dose and 28 receiving a high dose. Volumetric analysis of the stented segment and of both reference segments was performed. Baseline stent measurements and both reference measurements were similar among the groups. With increasing doses, there was a stepwise reduction in IH accumulation within the stented segment (31+/-22 mm3 in control, 18+/-15 mm3 in low dose, and 13+/-14 mm3 in high dose, P<0.001). Post hoc analysis showed less IH accumulation when low- and high-dose patients were compared with control (P=0.009 and P<0.001, respectively), but not when low-dose patients were compared with high-dose patients (P=0.2). Focal late malapposition was seen in 1 high-dose patient. With increasing doses, there was no significant change in the reference segments. CONCLUSIONS: Paclitaxel-coated stents are effective in reducing in-stent neointimal tissue proliferation in humans. They are not associated with edge restenosis or significant late malapposition. PMID- 12566360 TI - Vitamin E reduces progression of atherosclerosis in low-density lipoprotein receptor-deficient mice with established vascular lesions. AB - BACKGROUND: A growing body of evidence from animal studies supports the hypothesis that oxidative stress-mediated mechanisms play a central role in early atherogenesis. In contrast, clinical trials with antioxidant vitamins have not produced consistent results in humans with established atherosclerosis. METHODS AND RESULTS: Low-density lipoprotein receptor-deficient mice (LDLR KO) were fed a high-fat diet for 3 months to induce atheroma. At this time, 1 group of mice was euthanized for examination of atherosclerosis, and 2 other groups were randomized to receive high-fat diet either alone or supplemented with vitamin E for 3 additional months. At the end of the study, LDLR KO on a vitamin E-supplemented fat diet had decreased 8,12-iso-isoprostane (iP)F(2alpha)-VI and monocyte chemoattractant protein-1 levels, but increased nitric oxide levels compared with mice on placebo. No difference in lipid levels was observed between the 2 groups. Compared with baseline, placebo group had progression of atherosclerosis. In contrast, vitamin E-treated animals showed a significant reduction in progression of atherosclerosis. CONCLUSIONS: These results demonstrate that in LDLR KO, vitamin E supplementation reduces progression of established atherosclerosis by suppressing oxidative and inflammatory reactions and increasing nitric oxide levels. PMID- 12566361 TI - Prognostic significance of angiogenic growth factor serum levels in patients with acute coronary syndromes. AB - BACKGROUND: In patients with acute coronary syndromes, compensatory processes are initiated, including angiogenesis and endothelial regeneration of ruptured or eroded plaques. Angiogenic growth factors like vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and basic fibroblast growth factor (bFGF) are upregulated during ischemia. However, it is unknown whether their serum levels are related to clinical outcome. METHODS AND RESULTS: We measured VEGF, HGF, and bFGF levels in 1090 patients with acute coronary syndromes. Angiographic evaluation was performed at baseline as well as death, and nonfatal myocardial infarctions were recorded during 6-month follow-up. HGF and VEGF, but not bFGF, were significantly and independently associated with the patients' outcome. Patients with elevated VEGF serum levels suffered from adverse outcome (adjusted hazard ratio, 2.50 [1.52 to 4.82]; P=0.002). VEGF elevation was associated with evidence of ischemia and was a significant predictor of the effect of glycoprotein IIb/IIIa inhibition. In contrast, patients with high HGF levels had a significantly lower event rate compared with patients with low HGF levels (adjusted hazard ratio, 0.33 [0.21 to 0.51]; P<0.001). HGF levels did not correlate with evidence of ischemia and did not predict the effect of abciximab. Intriguingly, however, HGF levels significantly correlated with angiographically visible collateralization of the target vessel (22.4% versus 10.5%; P<0.001). CONCLUSIONS: The angiogenic growth factors VEGF and HGF are independent predictors of the patients' prognosis in acute coronary syndromes. Whereas VEGF elevation correlated with the evidence of myocardial ischemia and indicated an adverse outcome, HGF elevation was independent of ischemia and associated with improved collateralization as well as a favorable prognosis. PMID- 12566362 TI - Detecting acute coronary syndrome in the emergency department with cardiac magnetic resonance imaging. AB - BACKGROUND: Managing chest pain in the emergency department remains a challenge with current diagnostic strategies. We hypothesized that cardiac MRI could accurately identify patients with possible or probable acute coronary syndrome. METHODS AND RESULTS: The diagnostic performance of MRI was evaluated in a prospective study of 161 consecutive patients. Enrollment required 30 minutes of chest pain compatible with myocardial ischemia but an ECG not diagnostic of acute myocardial infarction. MRI was performed at rest within 12 hours of presentation and included perfusion, left ventricular function, and gadolinium-enhanced myocardial infarction detection. MRI was interpreted qualitatively but also analyzed quantitatively. The sensitivity and specificity, respectively, for detecting acute coronary syndrome were 84% and 85% by MRI, 80% and 61% by an abnormal ECG, 16% and 95% for strict ECG criteria for ischemia (ST depression or T-wave inversion), 40% and 97% for peak troponin-I, and 48% and 85% for a TIMI risk score > or =3. The MRI was more sensitive than strict ECG criteria for ischemia (P<0.001), peak troponin-I (P<0.001), and the TIMI risk score (P=0.004), and MRI was more specific than an abnormal ECG (P<0.001). Multivariate logistic regression analysis showed MRI was the strongest predictor of acute coronary syndrome and added diagnostic value over clinical parameters (P<0.001). CONCLUSIONS: Resting cardiac MRI exhibited diagnostic operating characteristics suitable for triage of patients with chest pain in the emergency department. Performed urgently to evaluate chest pain, MRI accurately detected a high fraction of patients with acute coronary syndrome, including patients with enzyme negative unstable angina. PMID- 12566363 TI - Identification of hibernating myocardium with quantitative intravenous myocardial contrast echocardiography: comparison with dobutamine echocardiography and thallium-201 scintigraphy. AB - BACKGROUND: There are currently no data on the accuracy of intravenous myocardial contrast echocardiography (MCE) in detecting myocardial hibernation in man and its comparative accuracy to dobutamine echocardiography (DE) or thallium 201 (Tl(201)) scintigraphy. METHODS AND RESULTS: Twenty patients with coronary artery disease and ventricular dysfunction underwent MCE 1 to 5 days before bypass surgery and repeat echocardiography at 3 to 4 months. Patients also underwent DE (n=18) and rest-redistribution Tl(201) tomography (n=16) before revascularization. MCE was performed using continuous Optison infusion (12 to 16 cc/h) with intermittent pulse inversion harmonics and incremental triggering (1:1 to 1:8). Myocardial contrast intensity (MCI) replenishment curves were constructed to derive quantitative MCE indices of blood velocity and flow. Recovery of function occurred in 38% of dysfunctional segments. MCE parameters of perfusion in hibernating myocardium were similar to segments with normal function and higher than dysfunctional myocardium without recovery of function (P<0.001). The best MCE parameter for predicting functional recovery was Peak MCIxbeta, an index of myocardial blood flow (area under the curve, 0.83). MCE parameters were higher in segments with contractile reserve and Tl(201) uptake > or =60% (P<0.05) and identified viable segments without contractile reserve by DE. The sensitivity of Peak MCIxbeta >1.5 dB/s for recovery of function was 90% and was similar to Tl(201) scintigraphy (92%) and any contractile reserve (80%); specificity was higher than for Tl(201) and DE (63%, 45%, and 54%, respectively; P<0.05). CONCLUSIONS: MCE with intravenous contrast identifies myocardial hibernation in humans. Prediction of viable myocardium with MCE is best using quantification of myocardial blood flow and provides improved accuracy compared with DE and Tl(201) scintigraphy. PMID- 12566364 TI - Intravascular ultrasound-guided balloon angioplasty compared with stent: immediate and 6-month results of the multicenter, randomized Balloon Equivalent to Stent Study (BEST). AB - BACKGROUND: Balloon angioplasty guided by intravascular ultrasound (IVUS) makes it possible to choose the balloon size according to the true vessel diameter and to detect suboptimal results requiring subsequent stent implantation. The Balloon Equivalent to STent (BEST) study aimed to assess whether this strategy would give the same results as systematic stenting. METHODS AND RESULTS: A total of 132 of 254 patients were randomized to IVUS-guided percutaneous transluminal coronary angioplasty (aggressive PTCA), and 122 were randomized to stenting (stent group). We hypothesized that a difference of <8% in the 6-month angiographic restenosis rate (primary end point) could be considered noninferior. The aggressive PTCA procedure was longer and had a greater use of contrast medium than stenting. In the aggressive PTCA group, crossover to stent was needed in 58 patients (44%). At 6 months, 20 of 119 patients (16.8+/-6.7%) in the aggressive PTCA group and 21 of 116 patients (18.1+/-7.0%) in the stent group had restenosis. The difference was 1.3%, with an upper limit of 95% confidence interval of 7.1% (ie, less than the noninferiority boundary). The in-stent restenosis rate was higher in the stent group (15.5% versus 5%; P=0.02). The differences in minimum lumen diameter, lumen cross-section area, and 1-year event rate were not significant. CONCLUSIONS: A strategy of IVUS-guided angioplasty with provisional stenting is feasible and safe. At the cost of a more complex procedure, it reduces the stent rate by half, with similar 6-month angiographic IVUS and clinical outcome compared with stent implantation. PMID- 12566365 TI - Effects of AGI-1067 and probucol after percutaneous coronary interventions. AB - BACKGROUND: AGI-1067, a metabolically stable modification of probucol, is an equipotent antioxidant to probucol but is pharmacologically distinct. In a multicenter trial, we studied whether AGI-1067 reduces restenosis assessed by intravascular ultrasound (IVUS) after percutaneous coronary intervention (PCI) compared with placebo and probucol used as a positive control. METHODS AND RESULTS: Two weeks before PCI, 305 patients were randomly assigned to 1 of 5 treatment groups: placebo, probucol 500 mg BID, or AGI-1067 70, 140, or 280 mg once daily. Patients were treated for 2 weeks before and 4 weeks after PCI. Baseline and 6-month follow-up IVUS were interpreted by a blinded core laboratory. Stents were used in 85% of patients. Luminal area at the PCI site at follow-up was 2.66+/-1.58 mm2 for placebo, 3.69+/-2.69 mm2 for probucol, 2.75+/ 1.76 mm2 for AGI-1067 70 mg, 3.17+/-2.26 mm2 for AGI-1067 140 mg, and 3.36+/-2.12 mm2 for AGI-1067 280 mg (P=0.02 for the dose-response relationship; P< or =0.05 for AGI-1067 280 mg and probucol versus placebo). There was a mean narrowing of 5.3 mm3 of reference segment lumen in the placebo group and an enlargement in the AGI-1067 140- and 280-mg groups at follow-up (P=0.05 for 140 mg). An increase in QTc interval >60 ms occurred in 4.8% of placebo patients, 17.4% of probucol patients, and 4.8%, 2.4%, and 2.5% of patients in the AGI-1067 groups (P=0.02). CONCLUSIONS: AGI-1067 and probucol reduce restenosis after PCI. In contrast to probucol, AGI-1067 did not cause prolongation of the QTc interval and improved lumen dimensions of reference segments, suggestive of a direct effect on atherosclerosis. PMID- 12566366 TI - TAXUS III Trial: in-stent restenosis treated with stent-based delivery of paclitaxel incorporated in a slow-release polymer formulation. AB - BACKGROUND: The first clinical study of paclitaxel-eluting stent for de novo lesions showed promising results. We performed the TAXUS III trial to evaluate the feasibility and safety of paclitaxel-eluting stent for the treatment of in stent restenosis (ISR). METHODS AND RESULTS: The TAXUS III trial was a single arm, 2-center study that enrolled 28 patients with ISR meeting the criteria of lesion length < or =30 mm, 50% to 99% diameter stenosis, and vessel diameter 3.0 to 3.5 mm. They were treated with one or more TAXUS NIRx paclitaxel-eluting stents. Twenty-five patients completed the angiographic follow-up at 6 months, and 17 of these underwent intravascular ultrasound (IVUS) examination. No subacute stent thrombosis occurred up to 12 months, but there was one late chronic total occlusion, and additional 3 patients showed angiographic restenosis. The mean late loss was 0.54 mm, with neointimal hyperplasia volume of 20.3 mm3. The major adverse cardiac event rate was 29% (8 patients; 1 non-Q-wave myocardial infarction, 1 coronary artery bypass grafting, and 6 target lesion revascularization [TLR]). Of the patients with TLR, 1 had restenosis in a bare stent implanted for edge dissection and 2 had restenosis in a gap between 2 paclitaxel-eluting stents. Two patients without angiographic restenosis underwent TLR as a result of the IVUS assessment at follow-up (1 incomplete apposition and 1 insufficient expansion of the stent). CONCLUSIONS: Paclitaxel-eluting stent implantation is considered safe and potentially efficacious in the treatment of ISR. IVUS guidance to ensure good stent deployment with complete coverage of target lesion may reduce reintervention. PMID- 12566367 TI - Short-term heart rate variability strongly predicts sudden cardiac death in chronic heart failure patients. AB - BACKGROUND: The predictive value of heart rate variability (HRV) in chronic heart failure (CHF) has never been tested in a comprehensive multivariate model using short-term laboratory recordings designed to avoid the confounding effects of respiration and behavioral factors. METHODS AND RESULTS: A multivariate survival model for the identification of sudden (presumably arrhythmic) death was developed with data from 202 consecutive patients referred between 1991 and 1995 with moderate to severe CHF (age 52+/-9 years, left ventricular ejection fraction 24+/-7%, New York Heart Association class 2.3+/-0.7; the derivation sample). Time and frequency-domain HRV parameters obtained from an 8' recording of ECG at baseline and during controlled breathing (12 to 15 breaths/min) were challenged against clinical and functional parameters. This model was then validated in 242 consecutive patients referred between 1996 and 2001 (validation sample). In the derivation sample, sudden death was independently predicted by a model that included low-frequency power (LFP) of HRV during controlled breathing < or =13 ms2 and left ventricular end-diastolic diameter > or =77 mm (relative risk [RR] 3.7, 95% CI 1.5 to 9.3, and RR 2.6, 95% CI 1.0 to 6.3, respectively). The derivation model was also a significant predictor in the validation sample (P=0.04). In the validation sample, LFP < or =11 ms2 during controlled breathing and > or =83 ventricular premature contractions per hour on Holter monitoring were both independent predictors of sudden death (RR 3.0, 95% CI 1.2 to 7.6, and RR 3.7, 95% CI 1.5 to 9.0, respectively). CONCLUSIONS: Reduced short-term LFP during controlled breathing is a powerful predictor of sudden death in patients with CHF that is independent of many other variables. These results refine the identification of patients who may benefit from prophylactic implantation of a cardiac defibrillator. PMID- 12566368 TI - Myocardial production of C-type natriuretic peptide in chronic heart failure. AB - BACKGROUND: C-type natriuretic peptide (CNP) is a vasodilator produced by the vascular endothelium. It shares structural and physiological properties with the cardiac hormones atrial natriuretic peptide and brain natriuretic peptide (BNP), but little is known about its pathophysiological role in chronic heart failure (CHF). We assessed the hypothesis that CNP is produced by the heart in patients with CHF. METHODS AND RESULTS: Myocardial CNP production was determined (difference in plasma levels between the aortic root and coronary sinus [CS]) in 9 patients undergoing right and left heart catheterization as part of their CHF assessment (all male, age 59+/-9 years; New York Heart Association class 2.2+/ 0.1; left ventricular ejection fraction 29+/-5%; creatinine 105+/-8 micro mol/L [all values mean+/-SEM]). BNP, established as originating from myocardium, was assessed from the same samples as a positive control. Analyses were performed by a blinded operator using a standard competitive radioimmunoassay kit (Peninsula Laboratories, Bachem Ltd UK). A step-up (29%) in plasma CNP concentration was found from the aorta to the CS (3.55+/-1.53 versus 4.59+/-1.54 pg/mL, respectively; P=0.035). The mean increase in CNP was 0.90+/-0.35 pg/mL (range 0.05 to 2.80 pg/mL). BNP levels increased by 57% from aorta to CS (86.0+/-20.5 versus 135.0+/-42.2 pg/mL; P=0.01). CS CNP levels correlated with mean pulmonary capillary wedge pressure (r=0.82, P=0.007). CONCLUSIONS: We have shown that CNP is produced by the heart in patients with CHF. Although further evaluation is required to define its full pathophysiological role in this condition, CNP may represent an important new local mediator in the heart. PMID- 12566369 TI - Differential regulation of natriuresis by 20-hydroxyeicosatetraenoic Acid in human salt-sensitive versus salt-resistant hypertension. AB - BACKGROUND: Twenty-hydroxyeicosatetraenoic acid (20-HETE) is a cytochrome P450 metabolite of arachidonic acid that produces vasoconstriction and inhibition of renal tubular sodium transport. In Dahl rats, a 20-HETE deficiency plays a role in salt-sensitive (SS) hypertension. In humans, there are no data on regulation of 20-HETE by salt intake or on a role for this compound in SS hypertension. METHODS AND RESULTS: Thirteen salt-resistant (SR) and 13 SS hypertensive subjects had urine 20-HETE excretion measured during salt-loading and depletion. In all patients, 20-HETE was 66.6% higher in the salt-replete (1.75+/-0.25 micro g/h) than in the salt-depleted state (1.05+/-0.16, P<0.003). There was no difference in 20-HETE excretion between SR and SS patients in either state of salt balance. In SR patients, sodium excretion during salt-loading correlated with 20-HETE (r=0.61, P<0.03) but not with blood pressure. In contrast, in SS patients, sodium excretion did not correlate with 20-HETE but did correlate with blood pressure (r=0.66, P<0.02). Finally, in the SS group only, there was a negative correlation between body mass index and 20-HETE excretion (r=-0.79, P<0.002) that was present during both salt-loading and depletion. CONCLUSIONS: We demonstrate for the first time that 20-HETE excretion is regulated by salt intake in hypertension. We find a disrupted relationship between sodium excretion and 20-HETE in SS patients, which results in dependence of their salt excretion on blood pressure and may be related to the magnitude of their obesity. We conclude that salt-sensitivity of blood pressure in essential hypertension may result from impairment of a natriuretic mechanism dependent on 20-HETE. PMID- 12566370 TI - Angiotensin-converting enzyme inhibition increases human vascular tissue-type plasminogen activator release through endogenous bradykinin. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibition potentiates the tissue type plasminogen activator (t-PA) response to exogenous bradykinin. This study tested the hypothesis that ACE inhibition increases endothelial t-PA release through endogenous bradykinin. METHODS AND RESULTS: We measured the effect of intra-arterial enalaprilat (5 micro g/min) on forearm blood flow (FBF) and net t PA release before and during intra-arterial infusion of bradykinin (25 to 400 ng/min) and methacholine (3.2 to 12.8 microg/min) in 24 smokers pretreated with bradykinin receptor antagonist HOE 140 (100 microg/kg intravenously) or vehicle. There was no specific effect of HOE 140 on FBF or forearm vascular resistance (FVR, 29.9+/-3.6 versus 29.7+/-3.6 mm Hg x mL(-1) x min(-1) x 100 mL(-1) after vehicle and HOE 140, respectively, P=0.956 between groups). Resting FVR decreased during enalaprilat compared with vehicle or HOE 140, but not compared with baseline, and the effect was similar in the 2 groups (22.0+/-2.7 and 24.1+/-2.9 mm Hg x mL(-1) x min(-1) x 100 mL(-1), respectively, P=0.610). In contrast, enalaprilat significantly increased resting net t-PA release (from 0.6+/-0.4 to 1.7+/-0.6 ng. min(-1) x 100 mL(-1), P=0.002); this effect was abolished by HOE 140 (0.1+/-0.3 ng x min(-1) x 100 mL(-1), P=0.036 versus enalaprilat alone). Enalaprilat increased the effect of exogenous bradykinin on FBF 60% (from 17.5+/ 2.5 to 28.1+/-4.0 mL. min(-1) x 100 mL(-1) during 100 ng/min bradykinin, P=0.001) and on t-PA release 14-fold (from 21.2+/-7.9 to 317.4+/-118.9 ng x min(-1) x 100 mL(-1), P=0.024). Enalaprilat increased the t-PA response to bradykinin to a greater extent than the FBF response, shifting the relationship between net t-PA release and FBF (P=0.005). HOE 140 blocked these effects. There was no effect of enalaprilat or HOE 140 on the FBF or t-PA response to methacholine. CONCLUSION: ACE inhibition increases constitutive endothelial t-PA release through endogenous bradykinin. PMID- 12566371 TI - Diverse effects of increasing lisinopril doses on lipid abnormalities in chronic nephropathies. AB - BACKGROUND: Dyslipidemia frequently complicates chronic nephropathies and increases the risk of renal and cardiovascular events. This might be ameliorated by drugs, such as angiotensin-converting enzyme inhibitors, which effectively reduce proteinuria. METHODS AND RESULTS: In this longitudinal study, we evaluated the extent to which uptitration of the ACE inhibitor lisinopril to maximum tolerated doses (median [range]: 30 [10 to 40] mg/d) ameliorated proteinuria and dyslipidemia in 28 patients with nondiabetic chronic nephropathies. Maximum lisinopril doses significantly and safely reduced proteinuria, serum total, LDL cholesterol, and triglycerides without substantially affecting serum HDL and renal hemodynamics. Proteinuria already decreased at 10 mg/d. Serum lipids progressively and dose-dependently decreased during uptitration to maximum doses. Reduction in total and LDL cholesterol correlated with increases in serum albumin/total protein concentration and oncotic pressure, peaked at lisinopril maximum doses, and persisted after treatment withdrawal. Despite less proteinuria reduction, hypercholesterolemia decreased more (and reflected the increase in serum albumin) in hypoalbuminemic than in normoalbuminemic patients who, despite more proteinuria reduction, had less decrease in cholesterol and no changes in serum albumin. Changes in serum triglycerides were independent of changes in serum proteins, were strongly correlated with lisinopril doses (r=-0.89, P=0.003) and recovered promptly after treatment withdrawal. Lisinopril was well tolerated, did not affect renal hemodynamics, and caused symptomatic, reversible hypotension in only two patients. CONCLUSIONS: In chronic nephropathies, angiotensin converting enzyme inhibitor uptitration to maximum tolerated doses safely ameliorated hypertriglyceridemia by a direct, dose-dependent effect, and hypercholesterolemia through amelioration of the nephrotic syndrome, particularly in patients with more severe hypoalbuminemia. PMID- 12566372 TI - Combination of a normal D-dimer concentration and a non-high pretest clinical probability score is a safe strategy to exclude deep venous thrombosis. AB - BACKGROUND: Serial ultrasonography is reliable for the diagnosis of deep venous thrombosis in symptomatic patients, but the low prevalence of thrombosis in this group renders the approach costly and inconvenient to patients. We studied the clinical validity of the combination of a pretest clinical probability score and a D-dimer test in the initial evaluation of patients suspected of deep venous thrombosis. METHODS AND RESULTS: Patients with a normal D-dimer concentration (<500 fibrin equivalent units [FEU] microg/L) and a non-high probability score (<3) had no further testing. Patients with a normal D-dimer concentration and a high probability score (> or =3) underwent one ultrasonogram. Serial ultrasonography was performed in patients with an abnormal D-dimer concentration. Patients were followed for 3 months. A total of 812 patients were evaluable for efficacy. Only 1 of 176 patients (0.6%; 95% CI, 0.02% to 3.1%) with a normal D dimer concentration and a non-high probability score developed thrombosis during follow-up. A normal D-dimer concentration and a high probability score were found in 39 patients; 3 of them (7.7%; 95% CI, 1.6% to 20.9%) had thrombosis at presentation, and one (2.8%; 95% CI, 0.07% to 14. 5%) developed pulmonary embolism during follow-up. In 306 of 597 patients (51.3%) with an abnormal D dimer concentration, thrombosis was detected by serial ultrasonography. Six patients (2.1%; 95% CI, 0.8% to 4. 4%) developed thrombosis during follow-up. No deaths due to thromboembolism occurred during follow-up. The total need for ultrasonography was reduced by 29%. CONCLUSION: The combination of a non-high pretest clinical probability score and a normal D-dimer concentration is a safe strategy to rule out deep venous thrombosis and to withhold anticoagulation. PMID- 12566373 TI - Matrix metalloproteinase-9 pretreatment level predicts intracranial hemorrhagic complications after thrombolysis in human stroke. AB - BACKGROUND: Matrix metalloproteinase (MMP) expression is related to blood brain barrier disruption after cerebral ischemia. Moreover, MMP inhibitors reduce hemorrhagic transformation (HT) after embolic ischemia in tissue plasminogen activator (t-PA)-treated animals. We aimed to correlate plasmatic MMP levels with the appearance of intracranial bleeding complications in stroke patients treated with t-PA. METHODS AND RESULTS: Serial MMP-2 and MMP-9 determinations were performed (ELISA, ng/mL) in 41 strokes involving the middle cerebral artery territory in patients who received t-PA within 3 hours of stroke onset. Blood samples were obtained at baseline (pretreatment) and at 12 and 24 hours after symptom onset. Hemorrhagic events were classified according to CT criteria (petechial hemorrhagic infarctions [HI, 1 to 2] and large parenchymal hemorrhages [PH, 1 to 2]). Brain CT scan was obtained at 48 hours or when a neurological worsening occurred. HT was present in 36.5% of the patients (24.4% HI and 12.1% PH). MMP-2 values were unrelated to any subtype of HT. The highest baseline MMP-9 level (normal range <97 ng/mL) corresponded to patients who later developed a PH (PH: 270.2+/-87.8, non-HT: 126.3+/-127.5, HI: 94.6+/-88.7; P=0.047). A graded response was found between mean baseline MMP-9 levels and the degree of bleeding (HI-1=37.4; HI-2=111.0; PH-1=202.5; PH-2=337.8). Baseline MMP-9 was the most powerful predictor of PH appearance in the multiple logistic regression model (OR= 9.62; CI 1.31 to 70.26; P=0.025). CONCLUSIONS: Baseline MMP-9 level predicts PH appearance after t-PA treatment. Therefore, we suggest that MMP determination may increase the safety profile for thrombolysis and, in the future, anti-MMP drugs might be combined with t-PA to prevent hemorrhagic complications. PMID- 12566374 TI - Nicotine strongly activates dendritic cell-mediated adaptive immunity: potential role for progression of atherosclerotic lesions. AB - BACKGROUND: Antigen-presenting cells (APCs) such as monocytes and dendritic cells (DCs) stimulate T-cell proliferation and activation in the course of adaptive immunity. This cellular interaction plays a role in the growth of atherosclerotic plaques. Nicotine has been shown to increase the growth of atherosclerotic lesions. Therefore, we investigated whether nicotine can stimulate APCs and their T cell-stimulatory capacity using human monocyte-derived DCs and murine bone marrow-derived DCs as APCs. METHODS AND RESULTS: Nicotine dose-dependently (10( 8) to 10(-4) mol/L) induced DC expression of costimulatory molecules (ie, CD86, CD40), MHC class II, and adhesion molecules (ie, LFA-1, CD54). Moreover, nicotine induced a 7.0-fold increase in secretion of the proinflammatory T(H)1 cytokine interleukin-12 by human DCs. These effects were abrogated by the nicotinic receptor antagonist alpha-bungarotoxin and mecamylamine, respectively. The effects of nicotine were mediated in part by the phosphorylation of the PI3 kinase downstream target Akt and the mitogen-activated kinases ERK and p38 MAPK. Nicotine-stimulated APCs had a greater capacity to stimulate T-cell proliferation and cytokine secretion, as documented by mixed lymphocyte reactions and ovalbumin specific assays with ovalbumin-transgenic DO10.11 mice. In a murine model of atherosclerosis, nicotine significantly enhanced the recruitment of DCs to atherosclerotic lesions in vivo. CONCLUSIONS: Nicotine activates DCs and augments their capacity to stimulate T-cell proliferation and cytokine secretion. These effects of nicotine may contribute to its influence on the progression of atherosclerotic lesions. PMID- 12566375 TI - LOX-1 mediates oxidized low-density lipoprotein-induced expression of matrix metalloproteinases in human coronary artery endothelial cells. AB - BACKGROUND: Oxidized LDL (ox-LDL) accumulation in the atherosclerotic region may enhance plaque instability. Both accumulation of ox-LDL and expression of its lectin-like receptor, LOX-1, have been shown in atherosclerotic regions. This study was designed to examine the role of LOX-1 in the modulation of metalloproteinases (MMP-1 and MMP-3) in human coronary artery endothelial cells (HCAECs). METHODS AND RESULTS: HCAECs were incubated with ox-LDL (10 to 80 micro g/mL) for 1 to 24 hours. Ox-LDL increased the expression of MMP-1 (collagenase) and MMP-3 (stromelysin-1) in a concentration- and time-dependent manner. Ox-LDL also increased collagenase activity. Ox-LDL did not significantly affect the expression of tissue inhibitors of metalloproteinases. Native LDL had no effect on the expression of MMPs. The effects of ox-LDL were mediated by its endothelial receptor, LOX-1, because pretreatment of HCAECs with a blocking antibody to LOX-1 (JTX92, 10 micro g/mL) prevented the expression of MMPs in response to ox-LDL (P<0.01). In parallel experiments, ox-LDL caused the activation of protein kinase C (PKC), which was inhibited by LOX-1 antibody. The PKC-beta isoform played a critical role in the expression of MMPs, because the PKC-beta inhibitor hispidin reduced ox-LDL-induced activation of PKC and the expression of MMPs. Other PKC subunits (alpha, gamma, and epsilon) did not affect the expression of MMPs. CONCLUSIONS: These findings indicate that ox-LDL, via LOX-1 activation, modulates the expression and activity of MMPs in HCAECs. In this process, activation of the PKC-beta subunit plays an important signaling role. PMID- 12566376 TI - Myocardial infarct expansion and matrix metalloproteinase inhibition. AB - BACKGROUND: A potential mechanism for left ventricular (LV) remodeling after myocardial infarction (MI) is activation of the matrix metalloproteinases (MMPs). This study examined the effects of MMP inhibition (MMPi) on regional LV geometry and MMP levels after MI. METHODS AND RESULTS: In pigs instrumented with radiopaque markers to measure regional myocardial geometry, MI was created by ligating the obtuse marginals of the circumflex artery. In the first study, pigs were randomized to MMPi (n=7; PD166793, 20 mg x kg(-1) x d(-1)) or MI only (n=7) at 5 days after MI, and measurements were performed at 2 weeks. Regional MI areas were equivalent at randomization and were increased in the MI-only group at 2 weeks after MI compared with the MMPi group. In the second study, pigs randomized to MMPi (n=9) or MI only (n=8) were serially followed up for 8 weeks. At 8 weeks after MI, LV end-diastolic dimension was lower with MMPi than in the MI-only group (4.7+/-0.1 versus 5.1+/-0.1 cm, P<0.05). Regional MI area was reduced with MMPi at 8 weeks after MI (1.3+/-0.1 versus 1.7+/-0.1 cm2, P<0.05). MMPi reduced ex vivo MMP proteolytic activity. In the MI region, membrane-type MMP levels were normalized and levels of the endogenous tissue inhibitor of MMPs (TIMP-1) were increased compared with normal levels with MMPi. These effects were not observed in the MI-only group. CONCLUSIONS: MMPi attenuated the degree of post-MI LV dilation and expansion of the infarct during the late phase of MI healing. In addition, exogenous MMPi caused region-specific modulation of certain MMP and TIMP species. PMID- 12566377 TI - Acetylsalicylic acid inhibits cell proliferation by involving transforming growth factor-beta. AB - BACKGROUND: Acetylsalicylic acid (ASA) inhibits cell proliferation. This may be mediated by transforming growth factor-beta (TGF-beta). TGF-beta directly stops cell proliferation, restrains cells in G(0), and inhibits the uptake of platelet derived growth factor and insulin-like growth factor. These effects are identical to those observed with ASA treatment. METHODS AND RESULTS: We cultured rat thoracic aorta vascular smooth muscle cells and measured cytotoxicity, cell proliferation, cell cycle, transcription of TGF-beta1, and concentration of TGF beta1 in supernatant medium. ASA dose-dependently restrained cells in G(0) phase with no cytotoxic effect and inhibited cell proliferation by 30.86%. Anti-TGF beta1 reversed this inhibition by 30.21%. However, ASA treatment decreased TGF beta1 transcription and had no significant effect on TGF-beta1 concentration. CONCLUSIONS: TGF-beta seems to play an important role in ASA-mediated inhibition of cell proliferation. Therefore, treatment with ASA prevents coronary disease not only by means of its antiplatelet properties but also by an important inhibition of plaque growth. This relationship between ASA and TGF-beta explains many other effects, such as cancer chemoprevention, immunomodulation, and wound healing. The aim of this study was to demonstrate this link. PMID- 12566378 TI - Altered myocardial thin-filament function in the failing Dahl salt-sensitive rat heart: amelioration by endothelin blockade. AB - BACKGROUND: Dahl salt-sensitive rats fed a high-salt diet develop compensated left ventricular hypertrophy followed by a transition to myocardial failure. We previously reported an increase in a troponin T isoform (TnT3) and a decrease in TnT phosphorylation in failing Dahl salt-sensitive rat hearts compared with low salt controls. The present study was undertaken to determine whether the thin filament plays a role in depression of the contractile machinery in this model. METHODS AND RESULTS: Native thin filaments (NTFs) were isolated intact from rats with compensated left ventricular hypertrophy and failing hearts and compared with age-matched controls. NTF velocity was measured as a function of free calcium in the in vitro motility assay. Maximal velocity was similar in all groups. However, NTFs from failing hearts demonstrated a reduction in calcium sensitivity compared with controls, as reflected in the pCa50 (5.88+/-0.05 versus 6.22+/-0.05, respectively, P<0.001). No difference in thin-filament motility (pCa50, V(max)) was observed in rats with compensated left ventricular hypertrophy compared with controls. Protein kinase A treatment of NTFs from control and failing hearts had no effect on thin-filament calcium sensitivity. However, the endothelin receptor blocker bosentan prevented the reduction in thin filament calcium sensitivity found in failing hearts. CONCLUSIONS: The thin filament is a key modulator of contractile performance in the transition to failure in the Dahl salt-sensitive rat model. The alteration in thin-filament function may be mediated by an endothelin-triggered pathway potentially affecting protein kinase C signaling. PMID- 12566379 TI - Burgeoning dilemmas in the management of diabetes and cardiovascular disease: rationale for the Bypass Angioplasty Revascularization Investigation 2 Diabetes (BARI 2D) Trial. AB - A paradoxical increase in mortality attributable to diabetes has occurred, particularly during the last decade, despite the overall decrease in mortality attributable to coronary artery disease in patients without diabetes. Insulin resistance with or without frank type 2 diabetes has emerged as a major determinant of accelerated coronary artery disease and its sequelae. The advent of insulin sensitizers enables clinicians to target treatment of insulin resistance, as well as hyperglycemia and dyslipidemia. The prevalence of diabetes in the United States is enormous and is increasing rapidly. Patients with diabetes respond less favorably to percutaneous coronary interventions and surgery compared with nondiabetic patients. These considerations led to the initiation of the Bypass Angioplasty Revascularization Investigation 2 Diabetes (BARI 2D) trial. It is designed to determine whether treatment targeted to attenuate insulin resistance can arrest or retard progression of coronary artery disease compared with treatment targeted to the same level of glycemic control with an insulin-providing approach. It is designed also to determine whether early revascularization reduces mortality and morbidity in patients with type 2 diabetes whose cardiac symptoms are mild and stable. Despite challenges in study design and enrollment, intensive follow-up, and the long duration of follow-up planned, the questions being addressed are compelling and seem to merit the effort. PMID- 12566380 TI - Images in cardiovascular medicine. Restrictive cardiomyopathy in familial amyloidosis TTR-Arg-50. PMID- 12566381 TI - American Heart Association guide for improving cardiovascular health at the community level: a statement for public health practitioners, healthcare providers, and health policy makers from the American Heart Association Expert Panel on Population and Prevention Science. PMID- 12566382 TI - Nicotine and its ways. PMID- 12566383 TI - Interaction of retinal bZIP transcription factor NRL with Flt3-interacting zinc finger protein Fiz1: possible role of Fiz1 as a transcriptional repressor. AB - NRL (neural retina leucine zipper) is a basic motif leucine zipper transcription factor of the Maf-subfamily. Multiple phosphorylated isoforms of NRL are detected specifically in rod photoreceptors. NRL regulates the expression of several rod specific genes, including rhodopsin and cGMP phosphodiesterase beta-subunit, in synergy with other transcription factors (e.g. the homeodomain protein CRX). Missense mutations in the human NRL gene are associated with autosomal dominant retinitis pigmentosa, whereas the loss of its function leads to rodless retina in Nrl-knockout mice that exhibit enhanced S-cone function. To further elucidate the molecular mechanism(s) underlying NRL-mediated transcriptional regulation, we used yeast two-hybrid screening to isolate NRL-interacting proteins in the retina and report the identification of Flt3-interacting zinc-finger protein, Fiz1. Interaction of Fiz1 and NRL-leucine zipper was validated by GST pulldown assays and co-immunoprecipitation from bovine retinal nuclear extracts. Fiz1 suppressed NRL- but not CRX-mediated transactivation of rhodopsin promoter activity in transiently transfected CV1 cells. The mRNA and the protein for both Fiz1 and its only other known interacting protein Flt3, a receptor tyrosine kinase, are expressed in the retina. Our results indicate potential cross-talk among signaling pathways in the retina and suggest that the function of NRL is modulated by its interaction with specific repressor proteins. PMID- 12566384 TI - Linkage of mild malaria to the major histocompatibility complex in families living in Burkina Faso. AB - Tumor necrosis factor alpha (TNF alpha) is thought to be a critical mediator of malaria fever, and mild malaria was previously reported to be linked to the MHC region containing the tumor necrosis factor alpha gene (TNF). Thirty-four families from Burkina Faso were analyzed to test for linkage between polymorphisms within the MHC region and mild malaria using the maximum-likelihood binomial (MLB) program. Two-point analysis indicated linkage of mild malaria to TNFd (LOD = 3.27; P = 5.44 x 10(-5)). Using multipoint analysis, we also found evidence for linkage of mild malaria to the MHC region, with a peak close to TNF (LOD = 3.86; P = 1.22 x 10(-5)). Our results support genes within the MHC region being involved in mild malaria. In particular, the genetic variation within TNF may influence susceptibility to mild malaria. Nevertheless, TNF-238, TNF-244 and TNF-308 polymorphisms are unlikely to explain linkage of mild malaria to the MHC region, and the causal mutations remain to be identified. PMID- 12566385 TI - Clinical and functional effects of a deletion in a COOH-terminal lumenal loop of the skeletal muscle ryanodine receptor. AB - We have identified a patient affected by a relatively severe form of central core disease (CCD), carrying a heterozygous deletion (amino acids 4863-4869) in the pore-forming region of the sarcoplasmic reticulum calcium release channel. The functional effect of this deletion was investigated (i) in lymphoblastoid cells from the affected patient and her mother, who was also found to harbour the mutation and (ii) in HEK293 cells expressing recombinant mutant channels. Lymphoblastoid cells carrying the RYR1 deletion exhibit an 'unprompted' calcium release from intracellular stores, resulting in significantly smaller thapsigargin-sensitive intracellular Ca(2+) stores, compared with lymphoblastoid cells from control individuals. Blocking the RYR1 with dantrolene restored the intracellular calcium stores to levels similar to those found in control cells. Single channel and [(3)H]ryanodine binding measurements of heterologously expressed mutant channels revealed a reduced ion conductance and loss of ryanodine binding and regulation by Ca(2+). Heterologous expression of recombinant RYR1 peptides and analysis of their membrane topology demonstrate that the deleted amino acids are localized in the lumenal loop connecting membrane-spanning segments M8 and M10. We provide evidence that a deletion in the lumenal loop of RYR1 alters channel function and causes CCD. PMID- 12566386 TI - Testicular cancer susceptibility in the 129.MOLF-Chr19 mouse strain: additive effects, gene interactions and epigenetic modifications. AB - Testicular germ cell tumors (TGCTs) are the most common solid cancers affecting young men. Although the evidence for genetic predisposition to TGCTs in humans is compelling, the genetic control of susceptibility is poorly understood. The 129S1/SvImJ (129/Sv) inbred strain of mice is an excellent model for studying TGCT susceptibility. We previously reported a new mouse strain, the 129.MOLF Chr19 chromosome substitution strain, which develops spontaneous TGCTs at a high frequency (70-80%) as compared with the much lower rate in the 129/Sv strain (5%). To characterize the genetic control of TGCT susceptibility, we created a panel of single- and double-congenic strains derived from 129.MOLF-Chr19. The frequency of TGCTs in these strains suggests that several genes with additive and epistatic effects located at distinct sites on chromosome 19 control susceptibility. However, an alternative interpretation involving epigenesis is based on a striking correlation between TGCT frequency and the length of the MOLF derived congenic segment, regardless of their chromosomal location on Chr 19 in each congenic strain. We also show that bilateral TGCT cases result from the coincidental co-occurrence of unilateral TGCTs rather than from the action of distinct genes that control susceptibility to bilateral versus unilateral TGCT cases. Finally, we propose that these TGCTs result from disrupted testicular and spermatogenic developmental programs. PMID- 12566387 TI - Constitutive knockout of Surf1 is associated with high embryonic lethality, mitochondrial disease and cytochrome c oxidase deficiency in mice. AB - We report here the creation of a constitutive knockout mouse for SURF1, a gene encoding one of the assembly proteins involved in the formation of cytochrome c oxidase (COX). Loss-of-function mutations of SURF1 cause Leigh syndrome associated with an isolated and generalized COX deficiency in humans. The murine phenotype is characterized by the following hallmarks: (1) high post-implantation embryonic lethality, affecting approximately 90% of the Surf1(-/-) individuals; (2) early-onset mortality of post-natal individuals; (3) highly significant deficit in muscle strength and motor performance; (4) profound and isolated defect of COX activity in skeletal muscle and liver, and, to a lesser extent, heart and brain; (5) morphological abnormalities of skeletal muscle, characterized by reduced histochemical reaction to COX and mitochondrial proliferation; (6) no obvious abnormalities in brain morphology, reflecting the virtual absence of overt neurological symptoms. These results indicate a function for murine Surf1 protein (Surf1p) specifically related to COX and recapitulate, at least in part, the human phenotype. This is the first mammalian model for a nuclear disease gene of a human mitochondrial disorder. Our model constitutes a useful tool to investigate the function of Surf1p, help understand the pathogenesis of Surf1p deficiency in vivo, and evaluate the efficacy of treatment. PMID- 12566388 TI - Identification of multiple loci for Alzheimer disease in a consanguineous Israeli Arab community. AB - We have observed an unusually high prevalence of dementia of the Alzheimer type (DAT) in Wadi Ara, an inbred Arab community in northern Israel comprising approximately 850 persons over the age of 60 years. Family studies revealed that more than one-third of the DAT cases are members of one hamula (tribal group) within Wadi Ara. To map chromosomal loci contributing to DAT susceptibility, we conducted a 10 cM scan in a series of five cases and five controls selected from this hamula. Markers from 18 chromosomal regions showed significant allelic association with DAT (P<0.05). Locations on chromosomes 2, 9 and 10 remained significant after testing additional affected and non-demented individuals. Significant associations were also observed for markers on chromosome 12 which overlap with a locus implicated in previous genome scans. Analysis of allele frequency distributions for 12 markers spanning 20 cM on chromosome 9 narrowed the possible location of an DAT susceptibility gene to a 13 cM interval between D9S157 and D9S259 (most significant result: P = 2.3 x 10(-7)). Analysis of 14 markers spanning 24 cM on chromosome 12 narrowed the possible location to a 14 cM interval distal to the LRP1 locus (most significant result: P = 1.3 x 10(-6)). Evidence for linkage on chromosome 9 stemmed primarily from excess homozygosity of marker alleles in cases compared with controls, suggesting that the gene at this location behaves in either a recessive or additive fashion. The unique characteristics of this community together with the emergent human genome data should allow for the rapid identification of DAT genes in these candidate regions. PMID- 12566389 TI - Differential binding of transcription factor E2F-2 to the endothelin-converting enzyme-1b promoter affects blood pressure regulation. AB - The endothelin-converting enzyme (ECE)-1 gene is a candidate for human blood pressure (BP) regulation and we report the identification of the new gene variants T-839G, C-338A, L75F, A677V and C+295T. Transient transfection of the reporter constructs containing the -338A allele showed an increase in promoter activity compared with the wild-type promoter. EMSA revealed the specific binding of E2F-2 to both ECE-1b promoter sequences, with the -338A allele being associated with an increased affinity to E2F-2 compared with -338C. The clinical relevance of this finding was analyzed in 704 hypertensive patients. In untreated hypertensive women, both the -338A and -839G alleles were significantly associated with ambulatory BP values. This study provides the first evidence of a link between the cell-cycle-associated E2F family and BP regulation via a component of the endothelin system. PMID- 12566390 TI - Interaction between blood pressure quantitative trait loci in rats in which trait variation at chromosome 1 is conditional upon a specific allele at chromosome 10. AB - We have used inbred and congenic rat strains in F(2) segregation studies to discover epistasis in a polygenic model of hypertension. Previously, we have found evidence that the presence of a blood pressure quantitative trait locus (QTL) on chromosome 1 is conditional upon the allele status of chromosome 10. To prove the existence of an epistatic interaction we have analyzed congenic strains for chromosome 1 and 10 carrying high blood pressure QTL alleles from the spontaneously hypertensive rat on a normotensive background of the Wistar-Kyoto (WKY) rat. Additionally, a double congenic strain was developed with both chromosome 1 and 10 high blood pressure QTL alleles on the WKY background. Analysis of variance for blood pressure phenotypes as determined by radiotelemetry showed a significant effect for chromosome 10 but not chromosome 1 QTL alleles and demonstrated a significant interaction between the two loci (P<0.05). The interaction accounted for 5 mmHg of blood pressure. Thus, the identification of epistasis is critical to the understanding of the quantitative nature of blood pressure genetics. PMID- 12566391 TI - Loss of CBP acetyltransferase activity by PHD finger mutations in Rubinstein Taybi syndrome. AB - Disruption of one copy of the human CREB binding protein (CBP or CREBBP) gene leads to the Rubinstein-Taybi syndrome (RTS), a developmental disorder characterized by retarded growth and mental functions, broad thumbs, broad big toes and typical facial abnormalities. The CREB binding protein (CBP) is an essential transcriptional coactivator for many different transcription factors. CBP has the intrinsic ability to acetylate histones and other proteins, which is regarded as an important step in transcription activation. In vitro studies have shown that this enzymatic activity critically depends on the integrity of a plant homeodomain (PHD)-type zinc finger in the HAT domain of CBP. We therefore investigated whether PHD finger mutations are present in RTS patients. Mutational analysis of 39 patients revealed eight novel heterozygous mutations in the HAT domain of CBP, one of which alters a conserved PHD finger amino acid (E1278K), while a second mutation deletes exon 22, which encodes the central region of the PHD finger. Functional analysis of these RTS-associated PHD finger mutants showed that they lacked in vitro acetyltransferase activity towards histones and CBP itself and displayed reduced coactivator function for the transcription factor CREB. Importantly, in EBV-transformed lymphoblastoid cells from the exon 22 deletion patient we found approximately 50% less endogenous CBP HAT activity. These findings therefore underscore the functional importance of the PHD finger in vivo and imply that reduction of CBP HAT activity, as exemplified here by disruption of the PHD finger, is sufficient to cause RTS. PMID- 12566392 TI - A recent polyploidy superimposed on older large-scale duplications in the Arabidopsis genome. AB - The Arabidopsis genome contains numerous large duplicated chromosomal segments, but the different approaches used in previous analyses led to different interpretations regarding the number and timing of ancestral large-scale duplication events. Here, using more appropriate methodology and a more recent version of the genome sequence annotation, we investigate the scale and timing of segmental duplications in Arabidopsis. We used protein sequence similarity searches to detect duplicated blocks in the genome, used the level of synonymous substitution between duplicated genes to estimate the relative ages of the blocks containing them, and analyzed the degree of overlap between adjacent duplicated blocks. We conclude that the Arabidopsis lineage underwent at least two distinct episodes of duplication. One was a polyploidy that occurred much more recently than estimated previously, before the Arabidopsis/Brassica rapa split and probably during the early emergence of the crucifer family (24-40 Mya). An older set of duplicated blocks was formed after the monocot/dicot divergence, and the relatively low level of overlap among these blocks indicates that at least some of them are remnants of a larger duplication such as a polyploidy or aneuploidy. PMID- 12566393 TI - Evolutionary implications of microbial genome tetranucleotide frequency biases. AB - We compared nucleotide usage pattern conservation for related prokaryotes by examining the representation of DNA tetranucleotide combinations in 27 representative microbial genomes. For each of the organisms studied, tetranucleotide usage departures from expectations (TUD) were shared between related organisms using both Markov chain analysis and a zero-order Markov method. Individual strains, multiple chromosomes, plasmids, and bacteriophages share TUDs within a species. TUDs varied between coding and noncoding DNA. Grouping prokaryotes based on TUD profiles resulted in relationships with important differences from those based on 16S rRNA phylogenies, which may reflect unequal rates of evolution of nucleotide usage patterns following divergence of particular organisms from a common ancestor. By both symmetrical tree distance and likelihood analysis, phylogenetic trees based on TUD profiles demonstrate a level of congruence with 16S rRNA trees similar to that of both RpoA and RecA trees. Congruence of these trees indicates that there exists phylogenetic signal in TUD patterns, most prominent in coding region DNA. Because relationships demonstrated in TUD-based analyses utilize whole genomes, they should be considered complementary to phylogenies based on single genetic elements, such as 16S rRNA. PMID- 12566394 TI - Genomic sequence and transcriptional profile of the boundary between pericentromeric satellites and genes on human chromosome arm 10p. AB - Contiguous finished sequence from highly duplicated pericentromeric regions of human chromosomes is needed if we are to understand the role of pericentromeric instability in disease, and in gene and karyotype evolution. Here, we have constructed a BAC contig spanning the transition from pericentromeric satellites to genes on the short arm of human chromosome 10, and used this to generate 1.4 Mb of finished genomic sequence. Combining RT-PCR, in silico gene prediction, and paralogy analysis, we can identify two domains within the sequence. The proximal 600 kb consists of satellite-rich pericentromerically duplicated DNA which is transcript poor, containing only three unspliced transcripts. In contrast, the distal 850 kb contains four known genes (ZNF248, ZNF25, ZNF33A, and ZNF37A) and up to 32 additional transcripts of unknown function. This distal region also contains seven out of the eight intrachromosomal duplications within the sequence, including the p arm copy of the approximately 250-kb duplication which gave rise to ZNF33A and ZNF33B. By sequencing orthologs of the duplicated ZNF33 genes we have established that ZNF33A has diverged significantly at residues critical for DNA binding but ZNF33B has not, indicating that ZNF33B has remained constrained by selection for ancestral gene function. These results provide further evidence of gene formation within intrachromosomal duplications, but indicate that recent interchromosomal duplications at this centromere have involved transcriptionally inert, satellite rich DNA, which is likely to be heterochromatic. This suggests that any novel gene structures formed by these interchromosomal events would require relocation to a more open chromatin environment to be expressed. PMID- 12566395 TI - Linkage disequilibrium and haplotype diversity in the genes of the renin angiotensin system: findings from the family blood pressure program. AB - Association studies of candidate genes with complex traits have generally used one or a few single nucleotide polymorphisms (SNPs), although variation in the extent of linkage disequilibrium (LD) within genes markedly influences the sensitivity and precision of association studies. The extent of LD and the underlying haplotype structure for most candidate genes are still unavailable. We sampled 193 blacks (African-Americans) and 160 whites (European-Americans) and estimated the intragenic LD and the haplotype structure in four genes of the renin-angiotensin system. We genotyped 25 SNPs, with all but one of the pairs spaced between 1 and 20 kb, thus providing resolution at small scale. The pattern of LD within a gene was very heterogeneous. Using a robust method to define haplotype blocks, blocks of limited haplotype diversity were identified at each locus; between these blocks, LD was lost owing to the history of recombination events. As anticipated, there was less LD among blacks, the number of haplotypes was substantially larger, and shorter haplotype segments were found, compared with whites. These findings have implications for candidate-gene association studies and indicate that variation between populations of European and African origin in haplotype diversity is characteristic of most genes. PMID- 12566396 TI - Sequence analysis of a functional Drosophila centromere. AB - Centromeres are the site for kinetochore formation and spindle attachment and are embedded in heterochromatin in most eukaryotes. The repeat-rich nature of heterochromatin has hindered obtaining a detailed understanding of the composition and organization of heterochromatic and centromeric DNA sequences. Here, we report the results of extensive sequence analysis of a fully functional centromere present in the Drosophila Dp1187 minichromosome. Approximately 8.4% (31 kb) of the highly repeated satellite DNA (AATAT and TTCTC) was sequenced, representing the largest data set of Drosophila satellite DNA sequence to date. Sequence analysis revealed that the orientation of the arrays is uniform and that individual repeats within the arrays mostly differ by rare, single-base polymorphisms. The entire complex DNA component of this centromere (69.7 kb) was sequenced and assembled. The 39-kb "complex island" Maupiti contains long stretches of a complex A+T rich repeat interspersed with transposon fragments, and most of these elements are organized as direct repeats. Surprisingly, five single, intact transposons are directly inserted at different locations in the AATAT satellite arrays. We find no evidence for centromere-specific sequences within this centromere, providing further evidence for sequence-independent, epigenetic determination of centromere identity and function in higher eukaryotes. Our results also demonstrate that the sequence composition and organization of large regions of centric heterochromatin can be determined, despite the presence of repeated DNA. PMID- 12566399 TI - Global RNA half-life analysis in Escherichia coli reveals positional patterns of transcript degradation. AB - Subgenic-resolution oligonucleotide microarrays were used to study global RNA degradation in wild-type Escherichia coli MG1655. RNA chemical half-lives were measured for 1036 open reading frames (ORFs) and for 329 known and predicted operons. The half-life of total mRNA was 6.8 min under the conditions tested. We also observed significant relationships between gene functional assignments and transcript stability. Unexpectedly, transcription of a single operon (tdcABCDEFG) was relatively rifampicin-insensitive and showed significant increases 2.5 min after rifampicin addition. This supports a novel mechanism of transcription for the tdc operon, whose promoter lacks any recognizable sigma binding sites. Probe by probe analysis of all known and predicted operons showed that the 5' ends of operons degrade, on average, more quickly than the rest of the transcript, with stability increasing in a 3' direction, supporting and further generalizing the current model of a net 5' to 3' directionality of degradation. Hierarchical clustering analysis of operon degradation patterns revealed that this pattern predominates but is not exclusive. We found a weak but highly significant correlation between the degradation of adjacent operon regions, suggesting that stability is determined by a combination of local and operon-wide stability determinants. The 16 ORF dcw gene cluster, which has a complex promoter structure and a partially characterized degradation pattern, was studied at high resolution, allowing a detailed and integrated description of its abundance and degradation. We discuss the application of subgenic resolution DNA microarray analysis to study global mechanisms of RNA transcription and processing. PMID- 12566397 TI - Centromere satellites from Arabidopsis populations: maintenance of conserved and variable domains. AB - The rapid evolution of centromere sequences between species has led to a debate over whether centromere activity is sequence-dependent. The Arabidopsis thaliana centromere regions contain approximately 20,000 copies of a 178-bp satellite repeat. Here, we analyzed satellites from 41 Arabidopsis ecotypes, providing the first broad population survey of satellite variation within a species. We found highly conserved segments and consistent sequence lengths in the Arabidopsis satellites and in the published collection of human alpha-satellites, supporting models for a functional role. Despite this conservation, polymorphisms are significantly enriched at some sites, yielding variation that could restrict binding proteins to a subset of repeat monomers. Some satellite regions vary considerably; at certain bases, consensus sequences derived from each ecotype diverge significantly from the Arabidopsis consensus, indicating substitutions sweep through a genome in less than 5 million years. Such rapid changes generate more variation within the set of Arabidopsis satellites than in genes from the chromosome arms or from the recombinationally suppressed centromere regions. These studies highlight a balance between the mechanisms that maintain particular satellite domains and the forces that disperse sequence changes throughout the satellite repeats in the genome. PMID- 12566398 TI - A microarray-based antibiotic screen identifies a regulatory role for supercoiling in the osmotic stress response of Escherichia coli. AB - Changes in DNA supercoiling are induced by a wide range of environmental stresses in Escherichia coli, but the physiological significance of these responses remains unclear. We now demonstrate that an increase in negative supercoiling is necessary for transcriptional activation of a large subset of osmotic stress response genes. Using a microarray-based approach, we have characterized supercoiling-dependent gene transcription by expression profiling under conditions of high salt, in conjunction with the microbial antibiotics novobiocin, pefloxacin, and chloramphenicol. Algorithmic clustering and statistical measures for gauging cellular function show that this subset is enriched for genes critical in osmoprotectant transport/synthesis and rpoS-driven stationary phase adaptation. Transcription factor binding site analysis also supports regulation by the global stress sigma factor rpoS. In addition, these studies implicate 60 uncharacterized genes in the osmotic stress regulon, and offer evidence for a broader role for supercoiling in the control of stress induced transcription. PMID- 12566400 TI - Bacillus subtilis during feast and famine: visualization of the overall regulation of protein synthesis during glucose starvation by proteome analysis. AB - Dual channel imaging and warping of two-dimensional (2D) protein gels were used to visualize global changes of the gene expression patterns in growing Bacillus subtilis cells during entry into the stationary phase as triggered by glucose exhaustion. The 2D gels only depict single moments during the cells' growth cycle, but a sequential series of overlays obtained at specific points of the growth curve facilitates visualization of the developmental processes at the proteomics scale. During glucose starvation a substantial reprogramming of the protein synthesis pattern was found, with 150 proteins synthesized de novo and cessation of the synthesis of almost 400 proteins. Proteins induced following glucose starvation belong to two main regulation groups: general stress/starvation responses induced by different stresses or starvation stimuli (sigma(B)-dependent general stress regulon, stringent response, sporulation), and glucose-starvation-specific responses (drop in glycolysis, utilization of alternative carbon sources, gluconeogenesis). Using the dual channel approach, it was not only possible to identify those regulons or stimulons, but also to follow the fate of each single protein by the three-color code: red, newly induced but not yet accumulated; yellow, synthesized and accumulated; and green, still present, but no longer being synthesized. These green proteins, which represent a substantial part of the protein pool in the nongrowing cell, are not accessible by using DNA arrays. The combination of 2D gel electrophoresis and MALDI TOF mass spectrometry with the dual channel imaging technique provides a new and comprehensive view of the physiology of growing or starving bacterial cell populations, here for the case of the glucose-starvation response. PMID- 12566401 TI - Coexpression of neighboring genes in Caenorhabditis elegans is mostly due to operons and duplicate genes. AB - In many eukaryotic species, gene order is not random. In humans, flies, and yeast, there is clustering of coexpressed genes that cannot be explained as a trivial consequence of tandem duplication. In the worm genome this is taken a step further with many genes being organized into operons. Here we analyze the relationship between gene location and expression in Caenorhabditis elegans and find evidence for at least three different processes resulting in local expression similarity. Not surprisingly, the strongest effect comes from genes organized in operons. However, coexpression within operons is not perfect, and is influenced by some distance-dependent regulation. Beyond operons, there is a relationship between physical distance, expression similarity, and sequence similarity, acting over several megabases. This is consistent with a model of tandem duplicate genes diverging over time in sequence and expression pattern, while moving apart owing to chromosomal rearrangements. However, at a very local level, nonduplicate genes on opposite strands (hence not in operons) show similar expression patterns. This suggests that such genes may share regulatory elements or be regulated at the level of chromatin structure. The central importance of tandem duplicate genes in these patterns renders the worm genome different from both yeast and human. PMID- 12566402 TI - Genome-scale reconstruction of the Saccharomyces cerevisiae metabolic network. AB - The metabolic network in the yeast Saccharomyces cerevisiae was reconstructed using currently available genomic, biochemical, and physiological information. The metabolic reactions were compartmentalized between the cytosol and the mitochondria, and transport steps between the compartments and the environment were included. A total of 708 structural open reading frames (ORFs) were accounted for in the reconstructed network, corresponding to 1035 metabolic reactions. Further, 140 reactions were included on the basis of biochemical evidence resulting in a genome-scale reconstructed metabolic network containing 1175 metabolic reactions and 584 metabolites. The number of gene functions included in the reconstructed network corresponds to approximately 16% of all characterized ORFs in S. cerevisiae. Using the reconstructed network, the metabolic capabilities of S. cerevisiae were calculated and compared with Escherichia coli. The reconstructed metabolic network is the first comprehensive network for a eukaryotic organism, and it may be used as the basis for in silico analysis of phenotypic functions. PMID- 12566403 TI - Transcriptional interactions between yeast tRNA genes, flanking genes and Ty elements: a genomic point of view. AB - Retroelement insertion can alter the expression of nearby genes. The Saccharomyces cerevisiae retrotransposons Ty1-Ty4 are transcribed by RNA polymerase II (pol II) and target their integration upstream of genes transcribed by RNA polymerase III (pol III), mainly tRNA genes. Because tRNA genes can repress nearby pol II-transcribed genes, we hypothesized that transcriptional interference may exist between Ty1 insertions and pol III-transcribed genes, the preferred targets for Ty1 integration. Ty1s upstream of two pol III-transcribed genes (SNR6 and SUP2) were recovered and analyzed by RNA blot analysis. Ty1 insertions were found to exert a neutral or modest stimulatory effect on the expression of these genes. Further RNA analysis indicated a modest tRNA position effect on Ty1 transcription. To investigate the possible genomic relevance of these expression effects, we compiled a comprehensive tRNA gene database. This database allowed us to analyze a genome's worth of tRNA genes and Ty elements. It also enabled the prediction and experimental confirmation of tRNA gene position effects at native chromosomal loci. We provide evidence supporting the hypothesis that tRNA genes exert a modest inhibitory effect on adjacent pol II promoters. Direct analysis of PTR3 transcription, promoted by sequences very close to a tRNA gene, shows that this tRNA position effect can operate on a native chromosomal gene. PMID- 12566404 TI - Systematic discovery of new genes in the Saccharomyces cerevisiae genome. AB - We used genome-wide comparative analysis of predicted protein sequences to identify many novel small genes, named smORFs for small open reading frames, within the budding yeast genome. Further analysis of 117 of these new genes showed that 84 are transcribed. We extended our analysis of one smORF conserved from yeast to human. This investigation provides an updated and comprehensive annotation of the yeast genome, validates additional concepts in the study of genomes in silico, and increases the expected numbers of coding sequences in a genome with the corresponding impact on future functional genomics and proteomics studies. PMID- 12566405 TI - Comparative genome analysis of the primary sex-determining locus in salmonid fishes. AB - We compared the Y-chromosome linkage maps for four salmonid species (Arctic charr, Salvelinus alpinus; Atlantic salmon, Salmo salar; brown trout, Salmo trutta; and rainbow trout, Oncorhynchus mykiss) and a putative Y-linked marker from lake trout (Salvelinus namaycush). These species represent the three major genera within the subfamily Salmoninae of the Salmonidae. The data clearly demonstrate that different Y-chromosomes have evolved in each of the species. Arrangements of markers proximal to the sex-determining locus are preserved on homologous, but different, autosomal linkage groups across the four species studied in detail. This indicates that a small region of DNA has been involved in the rearrangement of the sex-determining region. Placement of the sex-determining region appears telomeric in brown trout, Atlantic salmon, and Arctic charr, whereas an intercalary location for SEX may exist in rainbow trout. Three hypotheses are proposed to account for the relocation: translocation of a small chromosome arm; transposition of the sex-determining gene; or differential activation of a primary sex-determining gene region among the species. PMID- 12566406 TI - Complex events in the evolution of the human pseudoautosomal region 2 (PAR2). AB - The 320-kb human pseudoautosomal region 2 (PAR2) at the tips of the long arms of the X and Y chromosomes is thought to have been duplicated onto the Y chromosome recently in primate evolution. The four genes within PAR2 have been proposed to constitute two zones with different base ratios and transcription, one of which was added recently to the X chromosome. To test this hypothesis, we cloned and mapped PAR2 genes in other species, the lemur, the cat, and a marsupial, the tammar wallaby. None of the human PAR2 genes colocalized with human PAR1 genes in the marsupial genome, confirming that the human PAR1 and PAR2 evolved independently. Of the four PAR2 genes, only SYBL1 was located on the X chromosome in all species, including marsupials, so it was part of the ancient X. HSPRY3 localized to the X in all the eutherians, but not marsupial, so it must have been added to the X 80-130 million years ago. CXYorf1 was present on the X in primates and also in mouse, but autosomal in wallaby, suggesting a later addition 70-130 million years ago, and IL9R was on the X only in primate, suggesting addition 60 70 million years ago. The results therefore demonstrate that at least two independent additions were necessary for PAR2 evolution. The present gene order on the human X also requires two inversions. The complicated evolutionary pathway supports the hypothesis that terminal interchromosomal rearrangements are common in regions unpaired at meiosis. PMID- 12566407 TI - Polymorphism ratio sequencing: a new approach for single nucleotide polymorphism discovery and genotyping. AB - Polymorphism ratio sequencing (PRS) combines the advantages of high-throughput DNA sequencing with new labeling and pooling schemes to produce a powerful assay for sensitive single nucleotide polymorphism (SNP) discovery, rapid genotyping, and accurate, multiplexed allele frequency determination. In the PRS method, dideoxy-terminator extension ladders generated from a sample and reference template are labeled with different energy-transfer fluorescent dyes and coinjected into a separation capillary for comparison of relative signal intensities. We demonstrate the PRS method by screening two human mitochondrial genomes for sequence variations using a microfabricated capillary array electrophoresis device. A titration of multiplexed DNA samples places the limit of minor allele frequency detection at 5%. PRS is a sensitive and robust polymorphism detection method for the analysis of individual or multiplexed samples that is compatible with any four-color fluorescence DNA sequencer. PMID- 12566408 TI - Whole genome analysis of genetic alterations in small DNA samples using hyperbranched strand displacement amplification and array-CGH. AB - Structural genetic alterations in cancer often involve gene loss or gene amplification. With the advent of microarray approaches for the analysis of the genome, as exemplified by array-CGH (Comparative Genomic Hybridization), scanning for gene-dosage alterations is limited only by issues of DNA microarray density. However, samples of interest to the pathologist often comprise small clusters of just a few hundred cells, which do not provide sufficient DNA for array-CGH analysis. We sought to develop a simple method that would permit amplification of the whole genome without the use of thermocycling or ligation of DNA adaptors, because such a method would lend itself to the automated processing of a large number of tissue samples. We describe a method that permits the isothermal amplification of genomic DNA with high fidelity and limited sequence representation bias. The method is based on strand displacement reactions that propagate by a hyperbranching mechanism, and generate hundreds, or even thousands, of copies of the genome in a few hours. Using whole genome isothermal amplification, in combination with comparative genomic hybridization on cDNA microarrays, we demonstrate the ability to detect gene losses in yeast and gene dosage imbalances in human breast tumor cell lines. Although sequence representation bias in the amplified DNA presents potential problems for CGH analysis, these problems have been overcome by using amplified DNA in both control and tester samples. Gene-dosage alterations of threefold or more can be observed with high reproducibility with as few as 1000 cells of starting material. PMID- 12566409 TI - Identification and functional analysis of human transcriptional promoters. AB - Genomic and full-length cDNA sequences provide opportunities for understanding human gene structure and transcriptional regulatory elements. The simplest regulatory elements to identify are promoters, as their positions are dictated by the location of transcription start sites. We aligned full-length cDNA clones from the Mammalian Gene Collection to the human genome rough draft sequence to estimate the start sites of more than 10,000 human transcripts. We selected genomic sequence just upstream from the 5' end of these cDNA sequences and designated these as putative promoters. We assayed the functions of 152 of these DNA fragments, chosen at random from the entire set, in a luciferase-based transfection assay in four human cultured cell types. Ninety-one percent of these DNA fragments showed significant transcriptional activity in at least one of the cell lines, whereas 89% showed activity in at least two of the lines. We analyzed the distributions of strengths of these promoter fragments in the different cell types and identified likely alternative promoters in a large fraction of the genes. These data indicate that this approach is an effective method for predicting human promoters and provide the first set of functional data collected in parallel for a large set of human promoters. PMID- 12566410 TI - A complexity reduction algorithm for analysis and annotation of large genomic sequences. AB - DNA is a universal language encrypted with biological instruction for life. In higher organisms, the genetic information is preserved predominantly in an organized exon/intron structure. When a gene is expressed, the exons are spliced together to form the transcript for protein synthesis. We have developed a complexity reduction algorithm for sequence analysis (CRASA) that enables direct alignment of cDNA sequences to the genome. This method features a progressive data structure in hierarchical orders to facilitate a fast and efficient search mechanism. CRASA implementation was tested with already annotated genomic sequences in two benchmark data sets and compared with 15 annotation programs (10 ab initio and 5 homology-based approaches) against the EST database. By the use of layered noise filters, the complexity of CRASA-matched data was reduced exponentially. The results from the benchmark tests showed that CRASA annotation excelled in both the sensitivity and specificity categories. When CRASA was applied to the analysis of human Chromosomes 21 and 22, an additional 83 potential genes were identified. With its large-scale processing capability, CRASA can be used as a robust tool for genome annotation with high accuracy by matching the EST sequences precisely to the genomic sequences. PMID- 12566411 TI - Dynamic changes during the immune response in T cell-antigen-presenting cell clusters isolated from lymph nodes. AB - Activation of antigen-specific T cells by mature dendritic cells in secondary lymphoid organs is a key control point of the adaptive immune response. Here we describe the ex vivo isolation of preformed multicellular clusters between T cells and antigen-presenting cells. Adoptively transferred, antigen-specific T cells segregated into individual clusters where their activation and proliferation was initiated in vivo. Transit of the T cell cohort through the cluster compartment required 32-36 h. The precise timing of the response to agonistic epitopes was remarkably invariant regardless of the T cell lineage, the major histocompatibility complex haplotype, and the antigen dose. Interestingly, initiation of cell division of T cells specific for a subdominant epitope and a weak agonist was delayed by 6 h. The results provide a basis for the analysis of short range, mutual cell-cell interactions within such confined microenvironments. PMID- 12566413 TI - G-CSF-stimulated neutrophils are a prominent source of functional BLyS. AB - B lymphocyte stimulator (BLyS) is a novel member of the TNF ligand superfamily that is important in B cell maturation and survival. We demonstrate that human neutrophils, after incubation with G-CSF or, less efficiently, IFN gamma, express high levels of BLyS mRNA and release elevated amounts of biologically active BLyS. In contrast, surface expression of the membrane-bound BLyS was not detected in activated neutrophils. Indeed, in neutrophils, uniquely among other myeloid cells, soluble BLyS is processed intracellularly by a furin-type convertase. Worthy of note, the absolute capacity of G-CSF-stimulated neutrophils to release BLyS was similar to that of activated monocytes or dendritic cells, suggesting that neutrophils might represent an important source of BLyS. In this regard, we show that BLyS serum levels as well as neutrophil-associated BLyS are significantly enhanced after in vivo administration of G-CSF in patients. In addition, serum obtained from two of these patients induced a remarkable accumulation of neutrophil-associated BLyS in vitro. This effect was neutralized by anti-G-CSF antibodies, indicating that G-CSF, present in the serum, stimulated neutrophils to produce BLyS. Collectively, our findings suggest that neutrophils, through the production of BLyS, might play an unsuspected role in the regulation of B cell homeostasis. PMID- 12566412 TI - GATA-1 as a regulator of mast cell differentiation revealed by the phenotype of the GATA-1low mouse mutant. AB - Here it is shown that the phenotype of adult mice lacking the first enhancer (DNA hypersensitive site I) and the distal promoter of the GATA-1 gene (neo Delta HS or GATA-1(low) mutants) reveals defects in mast cell development. These include the presence of morphologically abnormal alcian blue(+) mast cells and apoptotic metachromatic(-) mast cell precursors in connective tissues and peritoneal lavage and numerous (60-70% of all the progenitors) "unique" trilineage cells committed to erythroid, megakaryocytic, and mast pathways in the bone marrow and spleen. These abnormalities, which were mirrored by impaired mast differentiation in vitro, were reversed by retroviral-mediated expression of GATA-1 cDNA. These data indicate an essential role for GATA-1 in mast cell differentiation. PMID- 12566414 TI - Regulation of lymphocyte apoptosis by interferon regulatory factor 4 (IRF-4). AB - To ensure that homeostasis of the immune system is maintained, the sensitivity of lymphocytes to Fas-mediated apoptosis is differentially regulated during their activation. The molecular mechanisms that link the activation program of lymphocytes to changes in sensitivity to Fas-mediated apoptosis have, however, not been fully characterized. In these studies, we have investigated whether Fas mediated apoptosis can be regulated by interferon regulatory factor 4 (IRF-4), a lymphoid-restricted member of the IRF family of transcription factors. IRF-4 expression is upregulated during lymphocyte activation and IRF-4-deficient mice have defects in both lymphocyte activation and homeostasis. Here, we show that stable expression of IRF-4 in a human lymphoid cell line that normally lacks IRF 4 leads to a significantly enhanced apoptotic response on Fas receptor engagement. A systematic examination of the downstream effectors of Fas signaling in IRF-4-transfected cells demonstrates an increased activation of caspase-8, as well as an increase in Fas receptor polarization. We demonstrate that IRF-4 deficient mice display defects in activation-induced cell death, as well as superantigen-induced deletion, and that these defects are accompanied by impairments in Fas receptor polarization. These data suggest that IRF-4, by modulating the efficiency of the Fas-mediated death signal, is a novel participant in the regulation of lymphoid cell apoptosis. PMID- 12566415 TI - The major histocompatibility complex-related Fc receptor for IgG (FcRn) binds albumin and prolongs its lifespan. AB - The inverse relationship between serum albumin concentration and its half-life suggested to early workers that albumin would be protected from a catabolic fate by a receptor-mediated mechanism much like that proposed for IgG. We show here that albumin binds FcRn in a pH dependent fashion, that the lifespan of albumin is shortened in FcRn-deficient mice, and that the plasma albumin concentration of FcRn-deficient mice is less than half that of wild-type mice. These results affirm the hypothesis that the major histocompatibility complex-related Fc receptor protects albumin from degradation just as it does IgG, prolonging the half-lives of both. PMID- 12566416 TI - Activation of dendritic cells through the interleukin 1 receptor 1 is critical for the induction of autoimmune myocarditis. AB - Dilated cardiomyopathy, resulting from myocarditis, is the most common cause of heart failure in young patients. We here show that interleukin (IL)-1 receptor type 1-deficient (IL-1R1(-/-)) mice are protected from development of autoimmune myocarditis after immunization with alpha-myosin-peptide(614-629). CD4(+) T cells from immunized IL-1R1(-/-) mice proliferated poorly and failed to transfer disease after injection into naive severe combined immunodeficiency (SCID) mice. In vitro stimulation experiments suggested that the function of IL-1R1(-/-)CD4(+) T cells was not intrinsically defect, but their activation by dendritic cells was impaired in IL-1R1(-/-) mice. Accordingly, production of tumor necrosis factor (TNF)-alpha, IL-1, IL-6, and IL-12p70 was reduced in dendritic cells lacking the IL-1 receptor type 1. In fact, injection of immature, antigen-loaded IL-1R1(+/+) but not IL-1R1(-/-) dendritic cells into IL-1R1(-/-) mice fully restored disease susceptibility by rendering IL-1R1(-/-) CD4(+) T cells pathogenic. Thus, IL-1R1 triggering is required for efficient activation of dendritic cells, which is in turn a prerequisite for induction of autoreactive CD4(+) T cells and autoimmunity. PMID- 12566417 TI - Extrathymic T cell lymphopoiesis: ontogeny and contribution to gut intraepithelial lymphocytes in athymic and euthymic mice. AB - In the absence of thymopoiesis, T lymphocytes are nevertheless present, mainly in the gut epithelium. Ontogeny of the extrathymic pathway and the extent of its involvement in euthymic mice are controversial. These questions have been addressed by assessing the expression of recombinase activating gene (RAG) through the use of green fluorescent protein RAG2 transgenic mouse models. In athymic mice, T lymphopoiesis occurs mainly in the mesenteric lymph node and less in the Peyer's patches. Ontogenic steps of this lymphopoiesis resemble those of thymopoiesis, but with an apparent bias toward gamma delta T cell production and with a paucity of oligoclonal alpha beta T cells possibly resulting from a deficit in positive selection. Whether in athymic or euthymic mice, neither T intraepithelial lymphocytes (IEL) nor cryptopatch cells (reported to contain precursors of IEL) displayed fluorescence indicating recent RAG protein synthesis. Newly made T cells migrate from the mesenteric node into the thoracic duct lymph to reach the gut mucosa. In euthymic mice, this extrathymic pathway is totally repressed, except in conditions of severe lymphocytic depletion. Thus, in normal animals, all gut T IEL, including CD8 alpha alpha(+) cells, are of thymic origin, CD8 alpha alpha(+) TCR alpha beta(+) IEL being the likely progeny of double negative NK1-1(-) thymocytes, which show polyclonal V alpha and V beta repertoires. PMID- 12566418 TI - The poxvirus protein A52R targets Toll-like receptor signaling complexes to suppress host defense. AB - Toll-like receptors (TLRs) are crucial in the innate immune response to pathogens, in that they recognize and respond to pathogen associated molecular patterns, which leads to activation of intracellular signaling pathways and altered gene expression. Vaccinia virus (VV), the poxvirus used to vaccinate against smallpox, encodes proteins that antagonize important components of host antiviral defense. Here we show that the VV protein A52R blocks the activation of the transcription factor nuclear factor kappa B (NF-kappa B) by multiple TLRs, including TLR3, a recently identified receptor for viral RNA. A52R associates with both interleukin 1 receptor-associated kinase 2 (IRAK2) and tumor necrosis factor receptor-associated factor 6 (TRAF6), two key proteins important in TLR signal transduction. Further, A52R could disrupt signaling complexes containing these proteins. A virus deletion mutant lacking the A52R gene was attenuated compared with wild-type and revertant controls in a murine intranasal model of infection. This study reveals a novel mechanism used by VV to suppress the host immunity. We demonstrate viral disabling of TLRs, providing further evidence for an important role for this family of receptors in the antiviral response. PMID- 12566419 TI - Integrin-dependence of lymphocyte entry into the splenic white pulp. AB - The steps involved in lymphocyte homing to the white pulp cords of the spleen are poorly understood. We demonstrate here that the integrins lymphocyte function associated (LFA)-1 and alpha 4 beta 1 make essential and mostly overlapping contributions necessary for B cell migration into white pulp cords. T cell entry to the white pulp is also reduced by blockade of LFA-1 and alpha 4 beta 1. The LFA-1 ligand, intercellular adhesion molecule 1 is critical for lymphocyte entry and both hematopoietic cells and radiation-resistant cells contribute to this requirement. Vascular cell adhesion molecule 1 contributes to the alpha 4 beta 1 ligand requirement and a second ligand, possibly fibronectin, also plays a role. By contrast with the entry requirements, antigen-induced movement of B cells from follicles to the outer T zone is not prevented by integrin blocking antibodies. Comparison of the distribution of integrin-blocked B cells and B cells treated with the G alpha i inhibitor, pertussis toxin, early after transfer reveals in both cases reduced accumulation in the inner marginal zone. These observations suggest that chemokine receptor signaling and the integrins LFA-1 and alpha 4 beta 1 function together to promote lymphocyte transit from the marginal zone into white pulp cords. PMID- 12566420 TI - Restricting Zap70 expression to CD4+CD8+ thymocytes reveals a T cell receptor dependent proofreading mechanism controlling the completion of positive selection. AB - Although T cell receptor (TCR) signals are essential for intrathymic T cell positive selection, it remains controversial whether they only serve to initiate this process, or whether they are required throughout to promote thymocyte differentiation and survival. To address this issue, we have devised a novel approach to interfere with thymocyte TCR signaling in a developmental stage specific manner in vivo. We have reconstituted mice deficient for Zap70, a tyrosine kinase required for TCR signaling and normally expressed throughout T cell development, with a Zap70 transgene driven by the adenosine deaminase (ADA) gene enhancer, which is active in CD4(+)CD8(+) thymocytes but inactive in CD4(+) or CD8(+) single-positive (SP) thymocytes. In such mice, termination of Zap70 expression impaired TCR signal transduction and arrested thymocyte development after the initiation, but before the completion, of positive selection. Arrested thymocytes had terminated Rag gene expression and up-regulated TCR and Bcl-2 expression, but failed to differentiate into mature CD4 or CD8 SP thymocytes, to be rescued from death by neglect or to sustain interleukin 7R alpha expression. These observations identify a TCR-dependent proofreading mechanism that verifies thymocyte TCR specificity and differentiation choices before the completion of positive selection. PMID- 12566421 TI - Second class minors: molecular identification of the autosomal H46 histocompatibility locus as a peptide presented by major histocompatibility complex class II molecules. AB - CD4 T cells regulate immune responses that cause chronic graft rejection and graft versus host disease but their target antigens remain virtually unknown. We developed a new method to identify CD4 T cell-stimulating antigens. LacZ inducible CD4 T cells were used as a probe to detect their cognate peptide/MHC II ligand generated in dendritic cells fed with Escherichia coli expressing a library of target cell genes. The murine H46 locus on chromosome 7 was thus found to encode the interleukin 4-induced IL4i1 gene. The IL4i1 precursor contains the HAFVEAIPELQGHV peptide which is presented by A(b) major histocompatibility complex class II molecule via an endogenous pathway in professional antigen presenting cells. Both allelic peptides bind A(b) and a single alanine to methionine substitution at p2 defines nonself. These results reveal novel features of H loci that regulate CD4 T cell responses as well as provide a general strategy for identifying elusive antigens that elicit CD4 T cell responses to tumors or self-tissues in autoimmunity. PMID- 12566422 TI - Immunotherapy of cytotoxic T cell-resistant tumors by T helper 2 cells: an eotaxin and STAT6-dependent process. AB - Currently most attempts at cancer immunotherapy involve the generation of CD8(+) cytotoxic T lymphocytes (CTLs) against tumor-associated antigens. Many tumors, however, have been immunoselected to evade recognition by CTLs and thus alternative approaches to cancer immunotherapy are urgently needed. Here we demonstrate that CD4(+) T cells that recognize a secreted tumor-specific antigen and exhibit a cytokine secretion profile characteristic of Th2 cells, are capable of clearing established lung and visceral metastases of a CTL-resistant melanoma. Clearance of lung metastases by the Th2 cells was found to be totally dependent on the eosinophil chemokine, eotaxin, and partially dependent on the transcription activator signal transducer and activator of transcription 6 (STAT6), with degranulating eosinophils within the tumors inducing tumor regression. In contrast, tumor-specific CD4(+) Th1 cells, that recruited macrophages into the tumors, had no effect on tumor growth. This work provides the basis for a new approach to adoptive T cell immunotherapy of cancer. PMID- 12566423 TI - LIS1 at the microtubule plus end and its role in dynein-mediated nuclear migration. AB - The cytoplasmic dynein complex and its accessory dynactin complex are involved in many cellular activities including nuclear migration in fungi (for review see Karki and Holzbaur, 1999). LIS1, the product of a causal gene for human lissencephaly (smooth brain), has also been implicated in dynein function based on studies in fungi and more recent studies in higher eukaryotic systems (for review see Gupta et al., 2002). Exactly how LIS1 may regulate the behavior of cytoplasmic dynein in various organisms is a fascinating question. In this issue, Lee et al. (2003) describe important new findings in Saccharomyces cerevisiae regarding the role of LIS1 (Pac1) in dynein-mediated nuclear migration. PMID- 12566424 TI - Dynactin polices two-way organelle traffic. AB - How is the bidirectional motion of organelles controlled? In this issue, Deacon et al. (2003) reveal the unexpected finding that dynactin (previously known to control dynein-based motility) binds to kinesin II and regulates anterograde movement of Xenopus melanosomes. This result suggests that dynactin may be a key player in coordinating vesicle traffic in this system. PMID- 12566425 TI - Anthrax toxin rafts into cells. AB - Anthrax toxin binds to a plasma membrane receptor and after endocytosis exerts its deadly effects on the cell. Until now, however, the mechanism of initial toxin uptake was unknown. In this issue, Abrami et al. (2003) demonstrate that toxin oligomerization clusters the anthrax receptor into lipid rafts and this complex is internalized via the clathrin-dependent pathway. PMID- 12566426 TI - The intramitochondrial dynamin-related GTPase, Mgm1p, is a component of a protein complex that mediates mitochondrial fusion. AB - A balance between fission and fusion events determines the morphology of mitochondria. In yeast, mitochondrial fission is regulated by the outer membrane associated dynamin-related GTPase, Dnm1p. Mitochondrial fusion requires two integral outer membrane components, Fzo1p and Ugo1p. Interestingly, mutations in a second mitochondrial-associated dynamin-related GTPase, Mgm1p, produce similar phenotypes to fzo1 and ugo cells. Specifically, mutations in MGM1 cause mitochondrial fragmentation and a loss of mitochondrial DNA that are suppressed by abolishing DNM1-dependent fission. In contrast to fzo1ts mutants, blocking DNM1-dependent fission restores mitochondrial fusion in mgm1ts cells during mating. Here we show that blocking DNM1-dependent fission in Deltamgm1 cells fails to restore mitochondrial fusion during mating. To examine the role of Mgm1p in mitochondrial fusion, we looked for molecular interactions with known fusion components. Immunoprecipitation experiments revealed that Mgm1p is associated with both Ugo1p and Fzo1p in mitochondria, and that Ugo1p and Fzo1p also are associated with each other. In addition, genetic analysis of specific mgm1 alleles indicates that Mgm1p's GTPase and GTPase effector domains are required for its ability to promote mitochondrial fusion and that Mgm1p self-interacts, suggesting that it functions in fusion as a self-assembling GTPase. Mgm1p's localization within mitochondria has been controversial. Using protease protection and immuno-EM, we have shown previously that Mgm1p localizes to the intermembrane space, associated with the inner membrane. To further test our conclusions, we have used a novel method using the tobacco etch virus protease and confirm that Mgm1p is present in the intermembrane space compartment in vivo. Taken together, these data suggest a model where Mgm1p functions in fusion to remodel the inner membrane and to connect the inner membrane to the outer membrane via its interactions with Ugo1p and Fzo1p, thereby helping to coordinate the behavior of the four mitochondrial membranes during fusion. PMID- 12566427 TI - The budding yeast Ipl1/Aurora protein kinase regulates mitotic spindle disassembly. AB - Ipl1p is the budding yeast member of the Aurora family of protein kinases, critical regulators of genomic stability that are required for chromosome segregation, the spindle checkpoint, and cytokinesis. Using time-lapse microscopy, we found that Ipl1p also has a function in mitotic spindle disassembly that is separable from its previously identified roles. Ipl1-GFP localizes to kinetochores from G1 to metaphase, transfers to the spindle after metaphase, and accumulates at the spindle midzone late in anaphase. Ipl1p kinase activity increases at anaphase, and ipl1 mutants can stabilize fragile spindles. As the spindle disassembles, Ipl1p follows the plus ends of the depolymerizing spindle microtubules. Many Ipl1p substrates colocalize with Ipl1p to the spindle midzone, identifying additional proteins that may regulate spindle disassembly. We propose that Ipl1p regulates both the kinetochore and interpolar microtubule plus ends to regulate its various mitotic functions. PMID- 12566428 TI - The role of the lissencephaly protein Pac1 during nuclear migration in budding yeast. AB - During mitosis in Saccharomyces cerevisiae, the mitotic spindle moves into the mother-bud neck via dynein-dependent sliding of cytoplasmic microtubules along the cortex of the bud. Here we show that Pac1, the yeast homologue of the human lissencephaly protein LIS1, plays a key role in this process. First, genetic interactions placed Pac1 in the dynein/dynactin pathway. Second, cells lacking Pac1 failed to display microtubule sliding in the bud, resulting in defective mitotic spindle movement and nuclear segregation. Third, Pac1 localized to the plus ends (distal tips) of cytoplasmic microtubules in the bud. This localization did not depend on the dynein heavy chain Dyn1. Moreover, the Pac1 fluorescence intensity at the microtubule end was enhanced in cells lacking dynactin or the cortical attachment molecule Num1. Fourth, dynein heavy chain Dyn1 also localized to the tips of cytoplasmic microtubules in wild-type cells. Dynein localization required Pac1 and, like Pac1, was enhanced in cells lacking the dynactin component Arp1 or the cortical attachment molecule Num1. Our results suggest that Pac1 targets dynein to microtubule tips, which is necessary for sliding of microtubules along the bud cortex. Dynein must remain inactive until microtubule ends interact with the bud cortex, at which time dynein and Pac1 appear to be offloaded from the microtubule to the cortex. PMID- 12566429 TI - Hierarchy of protein assembly at the vertex ring domain for yeast vacuole docking and fusion. AB - Vacuole tethering, docking, and fusion proteins assemble into a "vertex ring" around the apposed membranes of tethered vacuoles before catalyzing fusion. Inhibitors of the fusion reaction selectively interrupt protein assembly into the vertex ring, establishing a causal assembly hierarchy: (a) The Rab GTPase Ypt7p mediates vacuole tethering and forms the initial vertex ring, independent of t SNAREs or actin; (b) F-actin disassembly and GTP-bound Ypt7p direct the localization of other fusion factors; (c) The t-SNAREs Vam3p and Vam7p regulate each other's vertex enrichment, but do not affect Ypt7p localization. The v-SNARE Vti1p is enriched at vertices by a distinct pathway that is independent of the t SNAREs, whereas both t-SNAREs will localize to vertices when trans-pairing of SNAREs is blocked. Thus, trans-SNARE pairing is not required for SNARE vertex enrichment; and (d) The t-SNAREs regulate the vertex enrichment of both G-actin and the Ypt7p effector complex for homotypic fusion and vacuole protein sorting (HOPS). In accord with this hierarchy concept, the HOPS complex, at the end of the vertex assembly hierarchy, is most enriched at those vertices with abundant Ypt7p, which is at the start of the hierarchy. Our findings provide a unique view of the functional relationships between GTPases, SNAREs, and actin in membrane fusion. PMID- 12566430 TI - EPLIN regulates actin dynamics by cross-linking and stabilizing filaments. AB - Epithelial protein lost in neoplasm (EPLIN) is a cytoskeleton-associated protein encoded by a gene that is down-regulated in transformed cells. EPLIN increases the number and size of actin stress fibers and inhibits membrane ruffling induced by Rac. EPLIN has at least two actin binding sites. Purified recombinant EPLIN inhibits actin filament depolymerization and cross-links filaments in bundles. EPLIN does not affect the kinetics of spontaneous actin polymerization or elongation at the barbed end, but inhibits branching nucleation of actin filaments by Arp2/3 complex. Side binding activity may stabilize filaments and account for the inhibition of nucleation mediated by Arp2/3 complex. We propose that EPLIN promotes the formation of stable actin filament structures such as stress fibers at the expense of more dynamic actin filament structures such as membrane ruffles. Reduced expression of EPLIN may contribute to the motility of invasive tumor cells. PMID- 12566431 TI - Mechanism of filopodia initiation by reorganization of a dendritic network. AB - Afilopodium protrudes by elongation of bundled actin filaments in its core. However, the mechanism of filopodia initiation remains unknown. Using live-cell imaging with GFP-tagged proteins and correlative electron microscopy, we performed a kinetic-structural analysis of filopodial initiation in B16F1 melanoma cells. Filopodial bundles arose not by a specific nucleation event, but by reorganization of the lamellipodial dendritic network analogous to fusion of established filopodia but occurring at the level of individual filaments. Subsets of independently nucleated lamellipodial filaments elongated and gradually associated with each other at their barbed ends, leading to formation of cone shaped structures that we term Lambda-precursors. An early marker of initiation was the gradual coalescence of GFP-vasodilator-stimulated phosphoprotein (GFP VASP) fluorescence at the leading edge into discrete foci. The GFP-VASP foci were associated with Lambda-precursors, whereas Arp2/3 was not. Subsequent recruitment of fascin to the clustered barbed ends of Lambda-precursors initiated filament bundling and completed formation of the nascent filopodium. We propose a convergent elongation model of filopodia initiation, stipulating that filaments within the lamellipodial dendritic network acquire privileged status by binding a set of molecules (including VASP) to their barbed ends, which protect them from capping and mediate association of barbed ends with each other. PMID- 12566433 TI - 2'(3')-O-(N-methylanthraniloyl)-substituted GTP analogs: a novel class of potent competitive adenylyl cyclase inhibitors. AB - 2'(3')-O-(N-Methylanthraniloyl)-(MANT)-substituted nucleotides are fluorescent and widely used for the kinetic analysis of enzymes and signaling proteins. We studied the effects of MANT-guanosine 5'-[gamma-thio]triphosphate (MANT-GTP gamma S) and MANT-guanosine 5'-[beta,gamma-imido]triphosphate (MANT-GppNHp) on G alpha(s)- and G alpha(i)-protein-mediated signaling. MANT-GTP gamma S/MANT-GppNHp had lower affinities for G alpha(s) and G alpha(i) than GTP gamma S/GppNHp as assessed by inhibition of GTP hydrolysis of receptor-G alpha fusion proteins. MANT-GTP gamma S was much less effective than GTP gamma S at disrupting the ternary complex between the formyl peptide receptor and G alpha(i2). MANT-GTP gamma S/MANT-GppNHp non-competitively inhibited GTP gamma S/GppNHp-, AlF(4)(-)-, beta(2)-adrenoceptor plus GTP-, cholera toxin plus GTP-, and forskolin-stimulated adenylyl cyclase (AC) in G alpha(s)-expressing Sf9 insect cell membranes and S49 wild-type lymphoma cell membranes. AC inhibition by MANT-GTP gamma S/MANT-GppNHp was not due to G alpha(s) inhibition because it was also observed in G alpha(s) deficient S49 cyc(-) lymphoma cell membranes. Mn(2+) blocked AC inhibition by GTP gamma S/GppNHp in S49 cyc(-) membranes but enhanced the potency of MANT-GTP gamma S/MANT-GppNHp at inhibiting AC by approximately 4-8-fold. MANT-GTP gamma S and MANT-GppNHp competitively inhibited forskolin/Mn(2+)-stimulated AC in S49 cyc(-) membranes with K(i) values of 53 and 160 nm, respectively. The K(i) value for MANT-GppNHp at insect cell AC was 155 nm. Collectively, MANT-GTP gamma S/MANT GppNHp bind to G alpha(s)- and G alpha(i)-proteins with low affinity and are ineffective at activating G alpha. Instead, MANT-GTP gamma S/MANT-GppNHp constitute a novel class of potent competitive AC inhibitors. PMID- 12566432 TI - The ZO-1-associated Y-box factor ZONAB regulates epithelial cell proliferation and cell density. AB - Epithelial tight junctions regulate paracellular permeability, restrict apical/basolateral intramembrane diffusion of lipids, and have been proposed to participate in the control of epithelial cell proliferation and differentiation. Previously, we have identified ZO-1-associated nucleic acid binding proteins (ZONAB), a Y-box transcription factor whose nuclear localization and transcriptional activity is regulated by the tight junction-associated candidate tumor suppressor ZO-1. Now, we found that reduction of ZONAB expression using an antisense approach or by RNA interference strongly reduced proliferation of MDCK cells. Transfection of wild-type or ZONAB-binding fragments of ZO-1 reduced proliferation as well as nuclear ZONAB pools, indicating that promotion of proliferation by ZONAB requires its nuclear accumulation. Overexpression of ZONAB resulted in increased cell density in mature monolayers, and depletion of ZONAB or overexpression of ZO-1 reduced cell density. ZONAB was found to associate with cell division kinase (CDK) 4, and reduction of nuclear ZONAB levels resulted in reduced nuclear CDK4. Thus, our data indicate that tight junctions can regulate epithelial cell proliferation and cell density via a ZONAB/ZO-1-based pathway. Although this regulatory process may also involve regulation of transcription by ZONAB, our data suggest that one mechanism by which ZONAB and ZO-1 influence proliferation is by regulating the nuclear accumulation of CDK4. PMID- 12566434 TI - The molecular basis of the expansive substrate specificity of the antibiotic resistance enzyme aminoglycoside acetyltransferase-6'-aminoglycoside phosphotransferase-2". The role of ASP-99 as an active site base important for acetyl transfer. AB - The most frequent determinant of aminoglycoside antibiotic resistance in Gram positive bacterial pathogens is a bifunctional enzyme, aminoglycoside acetyltransferase-6'-aminoglycoside phosphotransferase-2" (AAC(6')- aminoglycoside phosphotransferase-2", capable of modifying a wide selection of clinically relevant antibiotics through its acetyltransferase and kinase activities. The aminoglycoside acetyltransferase domain of the enzyme, AAC(6') Ie, is the only member of the large AAC(6') subclass known to modify fortimicin A and catalyze O-acetylation. We have demonstrated through solvent isotope, pH, and site-directed mutagenesis effects that Asp-99 is responsible for the distinct abilities of AAC(6')-Ie. Moreover, we have demonstrated that small planar molecules such as 1-(bromomethyl)phenanthrene can inactivate the enzyme through covalent modification of this residue. Thus, Asp-99 acts as an active site base in the molecular mechanism of AAC(6')-Ie. The prominent role of this residue in aminoglycoside modification can be exploited as an anchoring site for the development of compounds capable of reversing antibiotic resistance in vivo. PMID- 12566435 TI - Inactivation of the hepatic cytochrome P450 system by conditional deletion of hepatic cytochrome P450 reductase. AB - Cytochrome P450 (CYP) monooxygenases catalyze the oxidation of a large number of endogenous compounds and the majority of ingested environmental chemicals, leading to their elimination and often to their metabolic activation to toxic products. This enzyme system therefore provides our primary defense against xenobiotics and is a major determinant in the therapeutic efficacy of pharmacological agents. To evaluate the importance of hepatic P450s in normal homeostasis, drug pharmacology, and chemical toxicity, we have conditionally deleted the essential electron transfer protein, NADH:ferrihemoprotein reductase (EC, cytochrome P450 reductase, CPR) in the liver, resulting in essentially complete ablation of hepatic microsomal P450 activity. Hepatic CPR-null mice could no longer break down cholesterol because of their inability to produce bile acids, and whereas hepatic lipid levels were significantly increased, circulating levels of cholesterol and triglycerides were severely reduced. Loss of hepatic P450 activity resulted in a 5-fold increase in P450 protein, indicating the existence of a negative feedback pathway regulating P450 expression. Profound changes in the in vivo metabolism of pentobarbital and acetaminophen indicated that extrahepatic metabolism does not play a major role in the disposition of these compounds. Hepatic CPR-null mice developed normally and were able to breed, indicating that hepatic microsomal P450-mediated steroid hormone metabolism is not essential for fertility, demonstrating that a major evolutionary role for hepatic P450s is to protect mammals from their environment. PMID- 12566436 TI - Low density lipoprotein receptor-related protein-mediated membrane translocation of 12/15-lipoxygenase is required for oxidation of low density lipoprotein by macrophages. AB - Oxidation of low density lipoprotein (LDL) is the key step for the development of atherosclerosis. The 12/15-lipoxygenase expressed in macrophages is capable of oxygenating linoleic acid esterified to cholesterol in the LDL particle, and thus this enzyme is presumed to initiate LDL oxidation. We recently reported that LDL receptor-related protein (LRP) was required for the enzyme-mediated LDL oxidation by macrophages and suggested the selective uptake of cholesterol ester from LDL to the plasma membrane (Xu, W., Takahashi, Y., Sakashita, T., Iwasaki, T., Hattori, H., and Yoshimoto. T. (2001) J. Biol. Chem. 276, 36454-36459). To elucidate precise mechanisms of lipoxygenase-mediated LDL oxidation, we investigated the intracellular localization of 12/15-lipoxygenase. The 12/15 lipoxygenase was predominantly detected in cytosol of resting peritoneal macrophages and of macrophage-like J774A.1 cells permanently transfected with the cDNA for the enzyme. When the cells were treated with LDL and subjected to subcellular fractionation, the 12/15-lipoxygenase was detected in the membranes with a concomitant decrease in cytosol as shown by Western blot analysis. The levels of the enzyme associated with the membrane reached maximum in 15 min after LDL addition and then decreased. However, the enzymatic activity of 12/15 lipoxygenase in the membrane fraction was very weak even after LDL treatment. This fact supports the suicide inactivation of the enzyme by the oxygenation of cholesterol ester transferred from the LDL particle to the plasma membrane. Immunohistochemical analysis using an antibody against 12/15-lipoxygenase revealed that the plasma membrane was the major site of the enzyme translocation by the LDL treatment. LDL-dependent 12/15-lipoxygenase translocation was inhibited by a blocking antibody against LRP. Furthermore, an enzyme translocation inhibitor, L655238, inhibited the LDL oxidation caused by the 12/15 lipoxygenase. We propose that cholesterol ester selectively transferred from the LDL particle to the plasma membrane via LRP is oxygenated by 12/15-lipoxygenase translocated to this membrane. PMID- 12566437 TI - Identification of regions of the tomato gamma-glutamyl kinase that are involved in allosteric regulation by proline. AB - The first step of proline biosynthesis is catalyzed by gamma-glutamyl kinase (GK). To better understand the feedback inhibition properties of GK, we randomly mutagenized a plasmid carrying tomato tomPRO1 cDNA, which encodes proline sensitive GK. A pool of mutagenized plasmids was transformed into an Escherichia coli GK mutant, and proline-overproducing derivatives were selected on minimal medium containing the toxic proline analog 3,4-dehydro-dl-proline. Thirty-two mutations that conferred 3,4-dehydro-dl-proline resistance were obtained. Thirteen different single amino acid substitutions were identified at nine different residues. The residues were distributed throughout the N-terminal two thirds of the polypeptide, but 9 mutations affecting 6 residues were in a cluster of 16 residues. GK assays revealed that these amino acid substitutions caused varying degrees of diminished sensitivity to proline feedback inhibition and also resulted in a range of increased proline accumulation in vivo. GK belongs to a family of amino acid kinases, and a predicted three-dimensional model of this enzyme was constructed on the basis of the crystal structures of three related kinases. In the model, residues that were identified as important for allosteric control were located close to each other, suggesting that they may contribute to the structure of a proline binding site. The putative allosteric binding site partially overlaps the dimerization and substrate binding domains, suggesting that the allosteric regulation of GK may involve a direct structural interaction between the proline binding site and the dimerization and catalytic domains. PMID- 12566438 TI - Biochemical properties of mammalian neutral sphingomyelinase 2 and its role in sphingolipid metabolism. AB - Neutral sphingomyelinase (N-SMase) is one of the key enzymes involved in the generation of ceramide; however, the gene(s) encoding for the mammalian N-SMase is still not well defined. Previous studies on the cloned nSMase1 had shown that the protein acts primarily as lyso-platelet-activating factor-phospholipase C. Recently the cloning of another putative N-SMase, nSMase2, was reported. In this study, biochemical characterization of the mouse nSMase2 was carried out using the overexpressed protein in yeast cells in which the inositol phosphosphingolipid phospholipase C (Isc1p) was deleted. N-SMase activity was dependent on Mg(2+) and was activated by phosphatidylserine and inhibited by GW4869. The ability of nSMase2 to recognize endogenous sphingomyelin (SM) as substrate was investigated by overexpressing nSMase2 in MCF7 cells. Mass measurements showed a 40% decrease in the SM levels in the overexpressor cells, and labeling studies demonstrated that nSMase2 accelerated SM catabolism. Accordingly, ceramide measurement showed a 60 +/- 15% increase in nSMase2 overexpressing cells compared with the vector-transfected MCF7. The role of nSMase2 in cell growth was next investigated. Stable overexpression of nSMase2 resulted in a 30-40% decrease in the rate of growth at the late exponential phase. Moreover, tumor necrosis factor induced approximately 50% activation of nSMase2 in MCF7 cells overexpressing the enzyme, demonstrating that nSMase2 is a tumor necrosis factor-responsive enzyme. In conclusion, these results 1) show that nSMase2 is a structural gene for nSMase, 2) suggest that nSMase2 acts as a bona fide N-SMase in cells, and 3) implicate nSMase2 in the regulation of cell growth and cell signaling. PMID- 12566439 TI - Sustained calcium entry through P2X nucleotide receptor channels in human airway epithelial cells. AB - Purinergic receptor stimulation has potential therapeutic effects for cystic fibrosis (CF). Thus, we explored roles for P2Y and P2X receptors in stably increasing [Ca(2+)](i) in human CF (IB3-1) and non-CF (16HBE14o(-)) airway epithelial cells. Cytosolic Ca(2+) was measured by fluorospectrometry using the fluorescent dye Fura-2/AM. Expression of P2X receptor (P2XR) subtypes was assessed by immunoblotting and biotinylation. In IB3-1 cells, ATP and other P2Y agonists caused only a transient increase in [Ca(2+)](i) derived from intracellular stores in a Na(+)-rich environment. In contrast, ATP induced an increase in [Ca(2+)](i) that had transient and sustained components in a Na(+) free medium; the sustained plateau was potentiated by zinc or increasing extracellular pH. Benzoyl-benzoyl-ATP, a P2XR-selective agonist, increased [Ca(2+)](i) only in Na(+)-free medium, suggesting competition between Na(+) and Ca(2+) through P2XRs. Biochemical evidence showed that the P2X(4) receptor is the major subtype shared by these airway epithelial cells. A role for store-operated Ca(2+) channels, voltage-dependent Ca(2+) channels, or Na(+)/Ca(2+) exchanger in the ATP-induced sustained Ca(2+) signal was ruled out. In conclusion, these data show that epithelial P2X(4) receptors serve as ATP-gated calcium entry channels that induce a sustained increase in [Ca(2+)](i). In airway epithelia, a P2XR mediated Ca(2+) signal may have therapeutic benefit for CF. PMID- 12566440 TI - Conserved motifs in somatostatin, D2-dopamine, and alpha 2B-adrenergic receptors for inhibiting the Na-H exchanger, NHE1. AB - Receptor subtypes within families of G protein-coupled receptors that are activated by similar ligands can regulate distinct intracellular effectors. We identified conserved motifs within intracellular domains 2 and 3 of selective subtypes of several G protein-coupled receptor families that confer coupling to the Na-H exchanger, NHE1. A T(s,p)V motif within intracellular domain 2 and a QQ(r) motif within intracellular domain 3 are shared by the somatostatin receptor subtypes SSTR1, -3, and -4, which couple to the inhibition of NHE1, but not by SSTR2 and -5, which do not signal to NHE1. Only the collective substitution of cognate SSTR2 residues with these two motifs conferred the ability of mutant SSTR2 to inhibit NHE1. Both motifs are present in D(2)-dopamine receptors, which inhibit NHE1, and in alpha(2B)-adrenergic receptors, which couple to the inhibition of NHE1, but not in alpha(2A)-adrenergic receptors, which do not regulate NHE1. These findings indicate that motifs shared by different subfamilies of G protein-coupled receptors, but not necessarily by receptor subtypes within a subfamily, can confer coupling to a common effector. PMID- 12566441 TI - Prostaglandin E2 induced functional expression of early growth response factor-1 by EP4, but not EP2, prostanoid receptors via the phosphatidylinositol 3-kinase and extracellular signal-regulated kinases. AB - Prostaglandin E(2) (PGE(2)) mediates its physiological effects by interactions with a subfamily of G-protein-coupled receptors known as EP receptors. These receptors consist of four primary subtypes named EP(1), EP(2), EP(3), and EP(4). The EP(2) and EP(4) subtypes are known to couple to Galpha(s) and stimulate intracellular cyclic 3,5- adenosine monophosphate formation, whereas the EP(1) and EP(3) receptors are known to couple to Galpha(q) and Galpha(i), respectively. Recently we found that EP(2) and EP(4) receptors can activate T-cell factor signaling; however, EP(2) receptors did this primarily through a cAMP-dependent protein kinase-dependent pathway, whereas EP(4) receptors primarily utilized a phosphatidylinositol 3-kinase (PI3K)-dependent pathway (Fujino, H., West, K. A., and Regan, J. W. (2002) J. Biol. Chem. 277, 2614-2619). We now report that PGE(2) stimulation of EP(4) receptors, but not EP(2) receptors, leads to phosphorylation of the extracellular signal-regulated kinases (ERKs) through a PI3K-dependent mechanism. Furthermore, this activation of PI3K/ERK signaling by the EP(4) receptors induces the functional expression of early growth response factor-1 (EGR-1). Under the same conditions induction of EGR-1 protein expression was not observed following PGE(2) stimulation of EP(2) receptors. These findings point to important differences in the signaling potential of the EP(2) and EP(4) receptors, which could be significant with respect to the potential involvement of EP(4) receptors in inflammation and cancer. PMID- 12566442 TI - A novel function of Rad54 protein. Stabilization of the Rad51 nucleoprotein filament. AB - Homologous recombination is important for the repair of double-stranded DNA breaks in all organisms. Rad51 and Rad54 proteins are two key components of the homologous recombination machinery in eukaryotes. In vitro, Rad51 protein assembles with single-stranded DNA to form the helical nucleoprotein filament that promotes DNA strand exchange, a basic step of homologous recombination. Rad54 protein interacts with this Rad51 nucleoprotein filament and stimulates its DNA pairing activity, suggesting that Rad54 protein is a component of the nucleoprotein complex involved in the DNA homology search. Here, using physical criteria, we demonstrate directly the formation of Rad54-Rad51-DNA nucleoprotein co-complexes that contain equimolar amounts of each protein. The binding of Rad54 protein significantly stabilizes the Rad51 nucleoprotein filament formed on either single-stranded DNA or double-stranded DNA. The Rad54-stabilized nucleoprotein filament is more competent in DNA strand exchange and acts over a broader range of solution conditions. Thus, the co-assembly of an interacting partner with the Rad51 nucleoprotein filament represents a novel means of stabilizing the biochemical entity central to homologous recombination, and reveals a new function of Rad54 protein. PMID- 12566443 TI - A ligand-induced conformational change in apolipoprotein(a) enhances covalent Lp(a) formation. AB - Lipoprotein(a) (Lp(a)) assembly proceeds via a two-step mechanism in which initial non-covalent interactions between apolipoprotein(a) (apo(a)) and low density lipoprotein precede disulfide bond formation. In this study, we used analytical ultracentrifugation, differential scanning calorimetry, and intrinsic fluorescence to demonstrate that in the presence of the lysine analog epsilon aminocaproic acid, apo(a) undergoes a substantial conformational change from a "closed" to an "open" structure that is characterized by an increase in the hydrodynamic radius (approximately 10%), an alteration in domain stability, as well as a decrease in tryptophan fluorescence. Although epsilon-aminocaproic acid is a well characterized inhibitor of the non-covalent interaction between apo(a) and low density lipoprotein, we report the novel observation that this ligand at low concentrations (100 microm-1 mm) significantly enhances covalent Lp(a) assembly by altering the conformation of apo(a). We developed a model for the kinetics of Lp(a) assembly that incorporates the conformational change as a determinant of the efficiency of the process; this model quantitatively explains our experimental observations. Interestingly, an analogous conformational change has been previously described for plasminogen resulting in an increase in the hydrodynamic radius, an increase in tryptophan fluorescence, and an acceleration of the rate of plasminogen activation. Although the functions of apo(a) and plasminogen have diverged considerably, elements of structural and conformational homology have been retained leading to similar regulation of two unrelated biological processes. PMID- 12566444 TI - Presentation of nitric oxide regulates monocyte survival through effects on caspase-9 and caspase-3 activation. AB - In the absence of survival factors, blood monocytes undergo spontaneous apoptosis, which involves the activation of caspase-3. Although nitric oxide can block caspase-3 activation and promote cell survival, it can also induce apoptosis. We hypothesized that nitrosothiols that promote protein S nitrosylation would reduce caspase-3 activation and cell survival, whereas nitric oxide donors (such as 1-propamine 3-(2-hydroxy-2-nitroso-1-propylhydrazine (PAPA) NONOate and diethylamine (DEA) NONOate) that do not target thiol residues would not. Using human monocytes as a model, we observed that nitrosothiol donors S nitrosoglutathione and S-nitroso-N-acetylpenicillamine suppressed caspase-9 and caspase-3 activity and DNA fragmentation. In contrast, PAPA or DEA NONOate did not promote monocyte survival events and appeared to inhibit monocyte survival induced by macrophage colony-stimulating factor. The caspase-3-selective inhibitor DEVD-fluoromethyl ketone reversed DNA fragmentation events, and the caspase-9 inhibitor LEHD-fluoromethyl ketone reversed caspase-3 activity in monocytes treated with PAPA or DEA NONOate in the presence of macrophage colony stimulating factor. These results were not caused by differences in glutathione levels or the kinetics of nitric oxide release. Moreover, S-nitrosoglutathione and S-nitroso-N-acetylpenicillamine directly blocked the activity of recombinant caspase-3, which was reversed by the reducing agent dithiothreitol, whereas PAPA or DEA NONOate did not block the enzymatic activity of caspase-3. These data support the hypothesis that nitrosylation of protein thiol residues by nitric oxide is critical for promoting the survival of human monocytes. PMID- 12566445 TI - Glucose metabolism in cancer. Evidence that demethylation events play a role in activating type II hexokinase gene expression. AB - One of the "signature" phenotypes of highly malignant, poorly differentiated tumors, including hepatomas, is their remarkable propensity to utilize glucose at a much higher rate than normal cells, a property frequently dependent on the marked overexpression of type II hexokinase (HKII). As the expression of the gene for this enzyme is nearly silent in liver tissue, we tested the possibility that DNA methylation/demethylation events may be involved in its regulation. Initial studies employing methylation restriction endonuclease analysis provided evidence for differential methylation patterns for the HKII gene in normal hepatocytes and hepatoma cells, the latter represented by a highly glycolytic model cell line (AS 30D). Subsequently, sequencing following sodium bisulfite treatment revealed 18 methylated CpG sites within a CpG island (-350 to +781 bp) in the hepatocyte gene but none in that of the hepatoma. In addition, treatment of a hepatocyte cell line with the DNA methyltransferase inhibitors, 5'-azacytidine and 5'-aza-2' deoxycytidine, activated basal expression levels of HKII mRNA and protein. Finally, stably transfecting the hepatocyte cell line with DNA demethylase also resulted in activating the basal expression levels of HKII mRNA and protein. These novel observations indicate that one of the initial events in activating the HKII gene during either transformation or tumor progression may reside at the epigenetic level. PMID- 12566446 TI - Involvement of the xenobiotic response element (XRE) in Ah receptor-mediated induction of human UDP-glucuronosyltransferase 1A1. AB - UDP-glucuronosyltransferase 1A1 (UGT1A1) plays an important physiological role by contributing to the metabolism of endogenous substances such as bilirubin in addition to xenobiotics and drugs. The UGT1A1 gene has been shown to be inducible by nuclear receptors steroid xenobiotic receptor (SXR) and the constitutive active receptor, CAR. In this report, we show that in human hepatoma HepG2 cells the UGT1A1 gene is also inducible with aryl hydrocarbon receptor (Ah receptor) ligands such as 2,3,7,8-tetrachlodibenzo-p-dioxin (TCDD), beta-naphthoflavone, and benzo[a]pyrene metabolites. Induction was monitored by increases in protein and catalytic activity as well as UGT1A1 mRNA. To examine the molecular interactions that control UGT1A1 expression, the gene was characterized and induction by Ah receptor ligands was regionalized to bases -3338 to -3287. Nucleotide sequence analysis of this UGT1A1 enhancer region revealed a xenobiotic response element (XRE) at -3381/-3299. The dependence of the XRE on UGT1A1 luciferase activity was demonstrated by a loss of Ah receptor ligand inducibility when the XRE core region (CACGCA) was deleted or mutated. Gel mobility shift analysis confirmed that TCDD induction of nuclear proteins specifically bound to the UGT1A1-XRE, and competition experiments with Ah receptor and Arnt antibodies demonstrated that the nuclear protein was the Ah receptor. These observations reveal that the Ah receptor is involved in human UGT1A1 induction. PMID- 12566448 TI - Post-translational secretion of fusion proteins in the halophilic archaea Haloferax volcanii. AB - Although protein secretion occurs post-translationally in bacteria and is mainly a cotranslational event in Eukarya, the relationship between the translation and translocation of secreted proteins in Archaea is not known. To address this question, the signal peptide-encoding region of the surface layer glycoprotein gene from the Haloarchaea Haloferax volcanii was fused either to the cellulose binding domain of the Clostridium thermocellum cellulosome or to the cytoplasmic enzyme dihydrofolate reductase from H. volcanii. Signal peptide-cleaved mature versions of both the cellulose-binding domain and dihydrofolate reductase could be detected in the growth medium of transformed H. volcanii cells. Immunoblot analysis revealed, however, the presence of full-length signal peptide-bearing forms of both proteins inside the cytoplasm of the transformed cells. Proteinase accessibility assays confirmed that the presence of cell-associated signal peptide-bearing proteins was not due to medium contamination. Moreover, the pulse radiolabeled signal peptide cellulose-binding domain chimera could be chased from the cytoplasm into the growth medium even following treatment with anisomycin, an antibiotic inhibitor of haloarchaeal protein translation. Thus, these results provide evidence that, in Archaea, at least some secreted proteins are first synthesized inside the cell and only then translocated across the plasma membrane into the medium. PMID- 12566447 TI - Identification of TIFA as an adapter protein that links tumor necrosis factor receptor-associated factor 6 (TRAF6) to interleukin-1 (IL-1) receptor-associated kinase-1 (IRAK-1) in IL-1 receptor signaling. AB - Tumor necrosis factor receptor-associated factor 6 (TRAF6) transduces signals from members of the Toll/interleukin-1 (IL-1) receptor family by interacting with IL-1 receptor-associated kinase-1 (IRAK-1) after IRAK-1 is released from the receptor-MyD88 complex upon IL-1 stimulation. However, the molecular mechanisms underlying regulation of the IRAK-1/TRAF6 interaction are largely unknown. We have identified TIFA, a TRAF-interacting protein with a forkhead-associated (FHA) domain. The FHA domain is a motif known to bind directly to phosphothreonine and phosphoserine. In transient transfection assays, TIFA activates NFkappaBeta and c Jun amino-terminal kinase. However, TIFA carrying a mutation that abolishes TRAF6 binding or mutations in the FHA domain that are known to abolish FHA domain binding to phosphopeptide fails to activate NFkappaBeta and c-Jun amino-terminal kinase. TIFA, when overexpressed, binds both TRAF6 and IRAK-1 and significantly enhances the IRAK-1/TRAF6 interaction. Furthermore, analysis of endogenous proteins indicates that TIFA associates with TRAF6 constitutively, whereas it associates with IRAK-1 in an IL-1 stimulation-dependent manner in vivo. Thus, TIFA is likely to mediate IRAK-1/TRAF6 interaction upon IL-1 stimulation. PMID- 12566449 TI - Mechanistic differences in the activation of estrogen receptor-alpha (ER alpha)- and ER beta-dependent gene expression by cAMP signaling pathway(s). AB - Although increases in intracellular cAMP can stimulate estrogen receptor-alpha (ER alpha) activity in the absence of exogenous hormone, no studies have addressed whether ER beta can be similarly regulated. In transient transfections, forskolin plus 3-isobutyl-1-methylxanthine (IBMX), which increases intracellular cAMP, stimulated the transcriptional activities of both ER alpha and ER beta. This effect was blocked by the protein kinase A inhibitor H89 (N-(2-(p bromocinnamylamino)-ethyl)-5-isoquinolinesulfonamide) and was dependent on an estrogen response element. A 12-O-tetradecanoylphorbol-13-acetate response element (TRE) located 5' to the estrogen response element was necessary for cAMP dependent activation of gene expression by ER beta but not ER alpha, indicating that the former subtype requires a functional interaction with TRE-interacting factor(s) to stimulate transcription. Both p160 and CREB-binding protein coactivators stimulated cAMP-induced ER alpha and ER beta transcriptional activity. However, mutation of the two cAMP-inducible SRC-1 phosphorylation sites important for cAMP activation of chicken progesterone receptor or all seven known SRC-1 phosphorylation sites did not specifically impair cAMP activation of ER alpha. The E/F domains of ER alpha are sufficient for activation by forskolin/IBMX, and this is accompanied by an increase in receptor phosphorylation. In contrast, cAMP signaling reduces the phosphorylation of the corresponding region of ER beta, and this correlates with the lack of forskolin/IBMX stimulated transcriptional activity. Our data suggest that cAMP activation of ER alpha transcriptional activity is associated with receptor instead of SRC-1 phosphorylation. Moreover, differences in the cofactor requirements, domains of ER alpha and ER beta sufficient for forskolin/IBMX activation, and the effect of cAMP on receptor phosphorylation indicate that this signaling pathway utilizes distinct mechanisms to stimulate ER alpha and ER beta transcriptional activity. PMID- 12566450 TI - Cross-regulation of novel protein kinase C (PKC) isoform function in cardiomyocytes. Role of PKC epsilon in activation loop phosphorylations and PKC delta in hydrophobic motif phosphorylations. AB - Recent studies identify conventional protein kinase C (PKC) isoform phosphorylations at conserved residues in the activation loop and C terminus as maturational events that influence enzyme activity and targeting but are not dynamically regulated by second messengers. In contrast, this study identifies phorbol 12-myristoyl 13-acetate (PMA)- and norepinephrine-induced phosphorylations of PKC epsilon (at the C-terminal hydrophobic motif) and PKC delta (at the activation loop) as events that accompany endogenous novel PKC (nPKC) isoform activation in neonatal rat cardiomyocytes. Agonist-induced nPKC phosphorylations are prevented (and the kinetics of PMA-dependent PKC down regulation are slowed) by pharmacologic inhibitors of nPKC kinase activity. PKC delta is recovered from PMA-treated cultures with increased in vitro lipid independent kinase activity (and altered substrate specificity); the PMA dependent increase in PKC delta kinase activity is attenuated when PKC delta activation loop phosphorylation is prevented. To distinguish roles of individual nPKC isoforms in nPKC phosphorylations, wild-type (WT) and dominant negative (DN) PKC delta and PKC epsilon mutants were introduced into cardiomyocyte cultures using adenovirus-mediated gene transfer. WT-PKC delta and WT-PKC epsilon are highly phosphorylated at activation loop and hydrophobic motif sites, even in the absence of allosteric activators. DN-PKC delta is phosphorylated at the activation loop but not the hydrophobic motif; DN-PKC epsilon is phosphorylated at the hydrophobic motif but not the activation loop. Collectively, these results identify a role for PKC epsilon in nPKC activation loop phosphorylations and PKC delta in nPKC hydrophobic motif phosphorylations. Agonist-induced nPKC isoform phosphorylations that accompany activation/translocation of the enzyme contribute to the regulation of PKC delta kinase activity, may influence nPKC isoform trafficking/down-regulation, and introduce functionally important cross-talk for nPKC signaling pathways in cardiomyocytes. PMID- 12566451 TI - Impaired NF-kappa B activation and increased production of tumor necrosis factor alpha in transgenic mice expressing keratin K10 in the basal layer of the epidermis. AB - Both the diversity and the precisely regulated tissue- and differentiation specific expression patterns of keratins suggest that these proteins have specific functions in epithelia besides their well known maintenance of cell integrity. In the search for these specific functions, our previous results have demonstrated that the expression of K10, a keratin expressed in postmitotic suprabasal cells of the epidermis, prevents cell proliferation through the inhibition of Akt kinase activity. Given the roles of Akt in NF-kappa B signaling and the importance of these processes in the epidermis, a study was made into the possible alterations of the NF-kappa B pathway in transgenic mice expressing K10 in the proliferative basal layer. It was found that the inhibition of Akt, mediated by K10 expression, leads to impaired NF-kappa B activity. This appears to occur through the decreased expression of IKK beta and IKK gamma. Remarkably, increased production of tumor necrosis factor alpha and concomitant JNK activation was observed in the epidermis of these transgenic mice. These results confirm that keratin K10 functions in vivo include the control of many aspects of epithelial physiology, which affect the cells not only in a cell autonomous manner but also influence tissue homeostasis. PMID- 12566452 TI - Pharmacological chaperone-mediated in vivo folding and stabilization of the P23H opsin mutant associated with autosomal dominant retinitis pigmentosa. AB - Protein conformational disorders, which include certain types of retinitis pigmentosa, are a set of inherited human diseases in which mutant proteins are misfolded and often aggregated. Many opsin mutants associated with retinitis pigmentosa, the most common being P23H, are misfolded and retained within the cell. Here, we describe a pharmacological chaperone, 11-cis-7-ring retinal, that quantitatively induces the in vivo folding of P23H-opsin. The rescued protein forms pigment, acquires mature glycosylation, and is transported to the cell surface. Additionally, we determined the temperature stability of the rescued protein as well as the reactivity of the retinal-opsin Schiff base to hydroxylamine. Our study unveils novel properties of P23H-opsin and its interaction with the chromophore. These properties suggest that 11-cis-7-ring retinal may be a useful therapeutic agent for the rescue of P23H-opsin and the prevention of retinal degeneration. PMID- 12566453 TI - The SNARE motif contributes to rbet1 intracellular targeting and dynamics independently of SNARE interactions. AB - The endoplasmic reticulum/Golgi SNARE rbet1 cycles between the endoplasmic reticulum and Golgi and is essential for cargo transport in the secretory pathway. Although the quaternary SNARE complex containing rbet1 is known to function in membrane fusion, the structural role of rbet1 is unclear. Furthermore, the structural determinants for rbet1 targeting and its cyclical itinerary have not been investigated. We utilized protein interaction assays to demonstrate that the rbet1 SNARE motif plays a structural role similar to the carboxyl-terminal helix of SNAP-25 in the synaptic SNARE complex and demonstrated the importance to SNARE complex assembly of a conserved salt bridge between rbet1 and sec22b. We also examined the potential role of the rbet1 SNARE motif and SNARE interactions in rbet1 localization and dynamics. We found that, in contrast to what has been observed for syntaxin 5, the rbet1 SNARE motif was essential for proper targeting. To test whether SNARE interactions were important for the targeting function of the SNARE motif, we used charge repulsion mutations at the conserved salt bridge position that rendered rbet1 defective for binary, ternary, and quaternary SNARE interactions. We found that heteromeric SNARE interactions are not required at any step in rbet1 targeting or dynamics. Furthermore, the heteromeric state of the SNARE motif does not influence its interaction with the COPI coat or efficient recruitment onto transport vesicles. We conclude that protein targeting is a completely independent function of the rbet1 SNARE motif, which is capable of distinct classes of protein interactions. PMID- 12566454 TI - Apparent cooperative assembly of the bacterial cell division protein FtsZ demonstrated by isothermal titration calorimetry. AB - The assembly dynamics of FtsZ, a prokaryotic homolog of tubulin, are important for their role in bacterial cytokinesis. Here we used isothermal titration calorimetry (ITC) to measure the heat of FtsZ self-association under various conditions. The measurements were designed to test whether FtsZ protofilaments are assembled by an isodesmic (linear aggregates in which each bond has an identical equilibrium constant) or a cooperative (aggregates only become stable after forming a oligomeric nucleus) assembly process. The isodesmic model can fit the assembly in GDP closely but cannot fit the assembly in GTP. FtsZ-GTP without Mg(2+) exhibits an apparent critical concentration, which is indicative of cooperative assembly, near 2.9 microm. With 2.5 mm Mg(2+) (which allows FtsZ to hydrolyze GTP) the critical concentration is reduced 10-fold to approximately 0.31 microm. Both with and without Mg(2+) there is no evidence for assembly below the critical concentration, but there is an abrupt transition to full assembly above. The ITC data are highly suggestive of a cooperative assembly, although this is difficult to reconcile with the 1-subunit-thick protofilaments observed by electron microscopy. PMID- 12566455 TI - Latrunculin B or ATP depletion induces cofilin-dependent translocation of actin into nuclei of mast cells. AB - Increasing cellular G-actin, using latrunculin B, in either intact or permeabilized rat peritoneal mast cells, caused translocation of both actin and an actin regulatory protein, cofilin, into the nuclei. The effect was not associated with an increase in the proportion of apoptotic cells. The major part of the nuclear actin was not stained by rhodamine-phalloidin but could be visualized with an actin antibody, indicating its monomeric or a conformationally distinct state, e.g. cofilin-decorated filaments. Introduction of anti-cofilin into permeabilized cells inhibited nuclear actin accumulation, implying that an active, cofilin-dependent, import exists in this system. Nuclear actin was localized outside the ethidium bromide-stained region, in the extrachromosomal nuclear domain. In permeabilized cells, the appearance of nuclear actin and cofilin was not significantly affected by increasing [Ca(2+)] and/or adding guanosine 5'-O-(3-thiotriphosphate), but was greatly promoted when ATP was withdrawn. Similarly, ATP depletion in intact cells also induced nuclear actin accumulation. In contrast to the effects of latrunculin B, ATP depletion was associated with an increase in cortical F-actin. Our results suggest that the presence of actin in the nucleus may be required for certain stress-induced responses and that cofilin is essential for the nuclear import of actin. PMID- 12566456 TI - Inhibition of Cdk4 activity enhances translation of p27kip1 in quiescent Rb negative cells. AB - We show in this work that the inhibition of Cdk4 (6) in Rb(-/-) 3T3 cells enhances the accumulation of the p27(kip1) cyclin-dependent kinase inhibitor when these cells are induced into quiescence. Two different forms of inhibition of Cdk4 (6), namely overexpression of the Cdk4 (6) inhibitor p16 and overexpression of a dominant negative mutant of Cdk4 (Cdk4(N158)), result in this effect. This suggests that the relevant activity of Cdk4 (6) that has to be inactivated in this setting is its kinase activity. The accumulation of p27(kip1) is due to enhanced translation of the protein, mediated by the 3'-untranslated region of the p27(kip1) mRNA. Moreover, the cells that overexpress p16(ink4a) or Cdk4(N158) show a delay in G(1) when made quiescent and restimulated to proliferate. This delay is overcome by transfection of a plasmid expressing antisense p27(kip1), which shows that the accumulation of p27(kip1) in these cells is related to their G(1) delay. In summary, we report a new functional link between two important cell cycle regulators, Cdk4 and p27(kip1), and provide a mechanistic explanation to the previously reported epistatic relations between these two proteins. PMID- 12566457 TI - Diketopyridylryanodine has three concentration-dependent effects on the cardiac calcium-release channel/ryanodine receptor. AB - By interacting with more than one site, ryanoids induce multiple effects on calcium-release channels. To date, the kinetics of interaction of only one of these sites has been characterized. Using C(4),C(12)-diketopyridylryanodine in both [(3)H]ryanodine binding and single channel experiments we characterized another site on the cardiac ryanodine receptor (RyR2) with which ryanoids interact. Competitive binding of this ryanoid to RyR2 implied a minimal two-site binding model. At the single channel level, C(4),C(12)-diketopyridylryanodine induced three distinct effects. At nanomolar concentrations, it increased channel open probability severalfold without inducing a subconductance. This effect was independent of membrane holding potential. As for other ryanoids, low micromolar concentrations of C(4),C(12)-diketopyridylryanodine readily induced a subconductance state. The major subconductance had a current amplitude of 52% of fully open, it was reversible, and its time to induction and duration were voltage- and concentration-dependent, affording Hill slopes of >2. At higher micromolar concentrations C(4),C(12)-diketopyridylryanodine induced long lasting, yet reversible shut states. Using a pharmacological strategy we have discerned an additional ryanoid-binding site on RyR2 that triggers an increase in channel activity. This site likely resides outside the strict confines of the transmembrane conducting pathway. PMID- 12566458 TI - The role of apoptosis signal-regulating kinase 1 in lymphotoxin-beta receptor mediated cell death. AB - LIGHT (homologous to lymphotoxins, shows inducible expression, and competes with herpes simplex virus glycoprotein D for herpesvirus entry mediator, a receptor expressed by T lymphocytes) is a member of the tumor necrosis factor superfamily that can interact with lymphotoxin-beta receptor (LTbetaR), herpes virus entry mediator, and decoy receptor (DcR3). In our previous study, we showed that LIGHT is able to induce cell death via the non-death domain containing receptor LTbetaR to activate both caspase-dependent and caspase-independent pathway. In this study, a LIGHT mutein, LIGHT-R228E, was shown to exhibit similar binding specificity as wild type LIGHT to LTbetaR, but lose the ability to interact with herpes virus entry mediator. By using both LIGHT-R228E and agonistic anti-LTbetaR monoclonal antibody, we found that signaling triggered by LTbetaR alone is sufficient to activate both caspase-dependent and caspase-independent pathways. Cross-linking of LTbetaR is able to recruit TRAF3 and TRAF5 to activate ASK1, whereas its activity is inhibited by free radical scavenger carboxyfullerenes. The activation of ASK1 is independent of caspase-3 activation, and kinase inactive ASK1-KE mutant can inhibit LTbetaR-mediated cell death. This suggests that ASK1 is one of the factors involved in the caspase-independent pathway of LTbetaR-induced cell death. PMID- 12566459 TI - Cdc42 is a Rho GTPase family member that can mediate insulin signaling to glucose transport in 3T3-L1 adipocytes. AB - We investigated the role of cdc42, a Rho GTPase family member, in insulin-induced glucose transport in 3T3-L1 adipocytes. Microinjection of anti-cdc42 antibody or cdc42 siRNA led to decreased insulin-induced and constitutively active G(q) (CA G(q); Q209L)-induced GLUT4 translocation. Adenovirus-mediated expression of constitutively active cdc42 (CA-cdc42; V12) stimulated 2-deoxyglucose uptake to 56% of the maximal insulin response, and this was blocked by treatment with the phosphatidylinositol 3-kinase (PI3-kinase) inhibitor, wortmannin, or LY294002. Both insulin and CA-G(q) expression caused an increase in cdc42 activity, showing that cdc42 is activated by insulin and is downstream of G alpha(q/11) in this activation pathway. Immunoprecipitation experiments showed that insulin enhanced a direct association of cdc42 and p85, and both insulin treatment and CA-cdc42 expression stimulated PI3-kinase activity in immunoprecipitates with anti-cdc42 antibody. Furthermore, the effects of insulin, CA-G(q), and CA-cdc42 on GLUT4 translocation or 2-deoxyglucose uptake were inhibited by microinjection of anti protein kinase C lambda (PKC lambda) antibody or overexpression of a kinase deficient PKC lambda construct. In summary, activated cdc42 can mediate 1) insulin-stimulated GLUT4 translocation and 2) glucose transport in a PI3-kinase dependent manner. 3) Insulin treatment and constitutively active G(q) expression can enhance the cdc42 activity state as well as the association of cdc42 with activated PI3-kinase. 4) PKC lambda inhibition blocks CA-cdc42, CA-G(q), and insulin-stimulated GLUT4 translocation. Taken together, these data indicate that cdc42 can mediate insulin signaling to GLUT4 translocation and lies downstream of G alpha(q/11) and upstream of PI3-kinase and PKC lambda in this stimulatory pathway. PMID- 12566460 TI - Sinorhizobium meliloti acpXL mutant lacks the C28 hydroxylated fatty acid moiety of lipid A and does not express a slow migrating form of lipopolysaccharide. AB - Lipid A is the hydrophobic anchor of lipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria. Lipid A of all Rhizobiaceae is acylated with a long fatty acid chain, 27-hydroxyoctacosanoic acid. Biosynthesis of this long acyl substitution requires a special acyl carrier protein, AcpXL, which serves as a donor of C28 (omega-1)-hydroxylated fatty acid for acylation of rhizobial lipid A (Brozek, K.A., Carlson, R.W., and Raetz, C. R. (1996) J. Biol. Chem. 271, 32126 32136). To determine the biological function of the C28 acylation of lipid A, we constructed an acpXL mutant of Sinorhizobium meliloti strain 1021. Gas-liquid chromatography and mass spectrometry analysis of the fatty acid composition showed that the acpXL mutation indeed blocked C28 acylation of lipid A. SDS-PAGE analysis of acpXL mutant LPS revealed only a fast migrating band, rough LPS, whereas the parental strain 1021 manifested both rough and smooth LPS. Regardless of this, the LPS of parental and mutant strains had a similar sugar composition and exposed the same antigenic epitopes, implying that different electrophoretic profiles might account for different aggregation properties of LPS molecules with and without a long acyl chain. The acpXL mutant of strain 1021 displayed sensitivity to deoxycholate, delayed nodulation of Medicago sativa, and a reduced competitive ability. However, nodules elicited by this mutant on roots of M. sativa and Medicago truncatula had a normal morphology and fixed nitrogen. Thus, the C28 fatty acid moiety of lipid A is not crucial, but it is beneficial for establishing an effective symbiosis with host plants. acpXL lies upstream from a cluster of five genes, including msbB (lpxXL), which might be also involved in biosynthesis and transfer of the C28 fatty acid to the lipid A precursor. PMID- 12566461 TI - Heparan sulfate regulates targeting of syndecan-1 to a functional domain on the cell surface. AB - In polarized B lymphoid cells, syndecan-1 is targeted specifically to a discrete membrane domain termed the uropod that is located at the cell's trailing edge. Within this functional domain, syndecan-1 promotes cell-cell adhesion and concentration of heparin binding growth factors. The present study reveals the surprising finding that targeting of syndecan-1 to uropods is mediated by its heparan sulfate chains and that targeting is regulated by cell surface events rather than solely by intracellular mechanisms. The addition of exogenous heparin or the treatment of polarized cells with heparitinase initiates a rapid and dramatic redistribution of uropod syndecan-1 over the entire cell surface, and a mutated syndecan-1 lacking heparan sulfate chains fails to concentrate within uropods. Interestingly, the heparan sulfate-bearing proteoglycans glypican-1 and beta glycan fail to concentrate in uropods, indicating that targeting may require heparan sulfate structural motifs unique to syndecan-1 or that the core protein of syndecan-1 participates in specific interactions that promote heparan sulfate mediated targeting. These findings suggest functional specificity for syndecan-1 within uropods and, in addition, reveal a novel mechanism for the targeting of molecules to discrete membrane subcellular domains via heparan sulfate. PMID- 12566462 TI - Regulation of pT alpha gene expression by a dosage of E2A, HEB, and SCL. AB - The expression of the pT alpha gene is required for effective selection, proliferation, and survival of beta T-cell receptor (beta TCR)-expressing immature thymocytes. Here, we have identified two phylogenetically conserved E boxes within the pT alpha enhancer sequence that are required for optimal enhancer activity and for its stage-specific activity in immature T cells. We have shown that the transcription factors E2A and HEB associate with high affinity to these E-boxes. Moreover, we have identified pT alpha as a direct target of E2A-HEB heterodimers in immature thymocytes because they specifically occupy the enhancer in vivo. In these cells, pT alpha mRNA levels are determined by the presence of one or two functional E2A or HEB alleles. Furthermore, E2A/HEB transcriptional activity is repressed by heterodimerization with SCL, a transcription factor that is turned off in differentiating thymocytes exactly at a stage when pT alpha is up-regulated. Taken together, our observations suggest that the dosage of E2A, HEB, and SCL determines pT alpha gene expression in immature T cells. PMID- 12566463 TI - Mapping the paratope of anti-CD4 recombinant Fab 13B8.2 by combining parallel peptide synthesis and site-directed mutagenesis. AB - We analyzed antigen-binding residues from the variable domains of anti-CD4 antibody 13B8.2 using the Spot method of parallel peptide synthesis. Sixteen amino acids, defined as Spot critical residues (SCR), were identified on the basis of a 50% decrease in CD4 binding to alanine analogs of reactive peptides. Recombinant Fab 13B8.2 mutants were constructed with alanine residues in place of each of the 16 SCR, expressed in the baculovirus cell system, and purified. CD measurements indicated that the mutated proteins were conformationally intact, with a beta-sheet secondary structure similar to that of wild-type Fab. Compared with the CD4-binding capacity of wild-type Fab 13B8.2, 11 light (Y32-L, W35-L, Y36-L, H91-L, and Y92-L) and heavy chain (H35-H, R38-H, W52-H, R53-H, F100K-H, and W103-H) Fab single mutants showed a decrease in CD4 recognition as demonstrated by enzyme-linked immunosorbent assay, BIAcore, and flow cytometry analyses. The five remaining Fab mutants showed antigen-binding properties similar to those of wild-type Fab. Recombinant Fab mutants that showed decreased CD4 binding also lost their capacity to inhibit human immunodeficiency virus promoter activation and the antigen-presenting ability that wild-type Fab displays. Molecular modeling of the 13B8.2 antibody paratope indicated that most of these critical residues are appropriately positioned inside the putative CD4 binding pocket, whereas the five SCR that were not confirmed by mutagenesis show an unfavorable positioning. Taken together, these results indicate that most of the residues defined by the Spot method as critical matched with important residues defined by mutagenesis in the whole protein context. The identification of critical residues for CD4 binding in the paratope of anti-CD4 recombinant Fab 13B8.2 provides the opportunity for the generation of improved anti-CD4 molecules with more efficient pharmacological properties. PMID- 12566465 TI - Zinc transporters 1, 2 and 4 are differentially expressed and localized in rats during pregnancy and lactation. AB - Zinc metabolism is controlled within relatively restricted limits throughout the life cycle. Expression and localization of zinc transporters 1, 2 and 4 during pregnancy and lactation in small intestine, mammary gland and liver of the rat were investigated using Northern analysis, Western blotting and immunohistochemistry. In maternal tissues, zinc transporter 4 was the most widely expressed among these zinc transporters in the tissues examined. In small intestine and liver, zinc transporter 4 increased from levels found during late gestation, but zinc transporter 1 did not. Zinc transporter 2 expression in small intestine was transient, being highest around parturition, and was not detected in liver. Immunohistochemistry revealed unique patterns of zinc transporter localization at different stages of development. In the placenta, zinc transporters 1 and 4 were found concentrated along the villous visceral splanchnopleure. In the mammary gland, zinc transporter 4 was most abundant in cells surrounding the alveolar ducts and oriented to the basement lamina. All three transporters were highly expressed in neonatal small intestine, principally near the apical surface, but zinc transporters 1 and 4 increased in abundance at the basolateral surface during development. Zinc transporter 2 was oriented apically, directly adjacent to the microvilli of enterocytes. Within the intestine, expression of each transporter was limited to enterocytes. These results support a role for these transporters in maintaining an adequate zinc supply derived from the maternal diet for zinc acquisition and use by the fetus and neonate. PMID- 12566466 TI - Soybean beta-conglycinin peptone suppresses food intake and gastric emptying by increasing plasma cholecystokinin levels in rats. AB - Cholecystokinin (CCK) is an important physiologic mediator that regulates satiety and gastric emptying. We demonstrated previously that soybean peptone acts directly on rat small intestinal mucosal cells to stimulate CCK release. In the present study, we examined the effects of beta-conglycinin, a major component of soy protein, and its peptone on food intake and gastric emptying after an intraduodenal infusion of beta-conglycinin peptone in relation to CCK release and interaction with the mucosal cell membrane. Intraduodenal infusion of beta conglycinin peptone inhibited food intake in a dose-dependent manner, but that of whole soy peptone or camostat did not. The suppression of food intake by beta conglycinin peptone was abolished by an intravenous injection of devazepide, a selective peripheral CCK receptor antagonist. The beta-conglycinin peptone infusion strongly suppressed gastric emptying with marked increases in portal CCK levels. We also observed that the beta-conglycinin peptone dose dependently and more potently stimulated CCK release from isolated dispersed mucosal cells of the rat jejunum than did beta-conglycinin itself. This stimulation corresponded to the binding activity of the peptide or protein to solubilized components of the rat jejunum membrane as evaluated by surface plasmon biosensor. These results indicate that beta-conglycinin peptone suppresses food intake, and this effect may be due to beta-conglycinin peptone in the lumen stimulating endogenous CCK release with direct acceptance to the intestinal cells. PMID- 12566467 TI - Increased translocation of cardiac protein kinase C beta2 accompanies mild cardiac hypertrophy in rats fed saturated fat. AB - Signal transduction through protein kinase C (PKC) beta2 may modulate cardiac hypertrophy in pressure-overloaded rat myocardium. Because PKC beta2 can be activated by fatty acids and diacylglycerol, we hypothesized that altering the level and type of dietary fat might modulate cardiac PKC activation and stimulate hypertrophy in otherwise normal rat myocardium. Male Sprague-Dawley rats (n = 32) were randomly assigned to either a low fat [10% total energy intake (TEI)] or high fat diet (40% TEI) based on corn or coconut oil as a source of saturated or unsaturated fat. After 40 d of isoenergetic diet consumption, the heart/body weight ratio was slightly greater in rats fed saturated fat diets compared with those fed unsaturated fat (P = 0.05). Increased activation of PKC beta2, as evidenced by greater membrane translocation, was also observed in all rats fed saturated fat diets (P < 0.01). PKC alpha, beta1 and epsilon did not change. These results suggest that dietary fat type can alter PKC beta2 activation in the heart, and exert a mild hypertrophic effect on the heart. PMID- 12566468 TI - Weaning and feed intake alter pancreatic enzyme activities and corresponding mRNA levels in 7-d-old piglets. AB - We investigated the changes in the capacity for synthesis of the exocrine pancreas of piglets during the 2 wk after weaning at 7 d of age (trial 1) by measuring the expression of digestive enzymes at mRNA and activity levels in pancreas homogenates, and the effects of high and low feed intakes during the 1st wk postweaning (trial 2) on these measures. The trypsin mRNA level was transiently decreased 43% 3 d postweaning (P < 0.05). Thereafter, trypsin and lipase mRNAs linearly increased (P < 0.05). During the 1st wk postweaning, trypsin- and lipase-specific activities were reduced 44 and 79% (P < 0.05), respectively, whereas 14 d after weaning, trypsin was at the preweaning value and lipase was at a low level. Amylase-specific activity did not change with weaning. Plasma cholecystokinin (CCK) and gastrin concentrations decreased 1 d postweaning and increased afterward up to 3 and 5 d postweaning, respectively. By 3 d after weaning, the mRNA level of trypsin was twofold higher (P < 0.05) in piglets that consumed more feed than in those that consumed less, whereas 7 d after weaning, the groups did not differ. By 7 d after weaning, the specific activity of amylase was higher, and lipase-specific activity was lower, in piglets that consumed more feed than in those that consumed less. Plasma CCK and gastrin concentrations measured 7 d after weaning were correlated with feed intake (r = +0.56 and r = +0.68, P < 0.05, respectively). In conclusion, by 3 d postweaning, pancreatic exocrine function was adapting to the new diet. Afterward, the expression of specific genes coding digestive enzymes and the levels of pancreatic enzyme activities were restored or stimulated, except for lipase-specific activity. Therefore, the pancreas can adjust to weaning and dry food intake as early as wk 2 of life. PMID- 12566469 TI - Refeeding and insulin regulate S6K1 activity in chicken skeletal muscles. AB - Broiler chickens are characterized by fast muscle growth and high protein deposition, most likely subsequent to a high protein synthesis. However, the regulation of protein synthesis in chicken muscle is still unknown. In contrast, it has been clearly demonstrated in mammals that S6K1 is a key regulator of protein synthesis. In the present study, S6K1 was characterized in both pectoralis and gastrocnemius muscles in chickens. A 133-bp fragment of chicken S6K1 cDNA had 84% identity to mammalian S6K1. We investigated in vivo the effects of refeeding and insulin treatment after 16 h starvation. S6K1 enzyme activity was significantly increased in both pectoralis and gastrocnemius muscles by refeeding (two- to threefold greater than in food-deprived chickens, P < 0.05). Optimal activation occurred 30 min after refeeding following 16 h starvation. S6K1 activation was associated with its phosphorylation on serine and Thr 389 residues, which occurred within the first 5 min of refeeding. S6K1 was also significantly stimulated in both pectoralis and gastrocnemius muscles after a single insulin injection (nine- to 12-fold greater than in control chickens, P < 0.001). Our results indicate that S6K1 is expressed in chickens muscles and activated by refeeding and insulin treatment. PMID- 12566470 TI - Vitamin D receptor (VDR) knockout mice reveal VDR-independent regulation of intestinal calcium absorption and ECaC2 and calbindin D9k mRNA. AB - To study the role of calbindin D(9k) (CaBP) and epithelial calcium channel ECaC2 in intestinal calcium (Ca) absorption, vitamin D receptor knockout (KO) and wild type (WT) mice were fed either 0.5% Ca or a 2.0% Ca rescue diet starting at 21 d of age. Ca absorption and parameters involved in this process were measured at 60 or 90 d of age. Compared with WT, KO mice fed the 0.5% Ca diet had higher plasma parathyroid hormone (PTH) and 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], and lower plasma Ca and insulin-like growth factor-I (IGF-I). Duodenal Ca absorption (% Ca absorbed) in KO mice was reduced 71% relative to WT mice and was associated with 55% lower CaBP mRNA, 47% lower CaBP protein and 95% lower ECaC2 mRNA levels. Compared with WT mice, the percentage of Ca absorbed in KO mice fed the 0.5% Ca diet was inappropriately low for the level of duodenal CaBP. The 2% Ca rescue diet normalized plasma Ca, prevented osteomalacia, increased growth and plasma IGF-I levels, but did not normalize plasma PTH or 1,25(OH)(2)D(3) in KO mice. In addition, the relationship between CaBP protein and the percentage of Ca absorbed was normalized, whereas ECaC2 mRNA fell to near zero. Our data demonstrate that higher CaBP levels do not ensure high rates of duodenal Ca absorption and that transcellular Ca absorption can occur even when ECaC2 gene expression is very low. In addition, our data suggest that the 2% Ca diet promotes a vitamin D receptor-independent anabolic effect on bone formation and calcium absorption, leading to improved calcium balance even in the presence of high PTH levels. PMID- 12566471 TI - Beta-carotene regulates NF-kappaB DNA-binding activity by a redox mechanism in human leukemia and colon adenocarcinoma cells. AB - We demonstrated previously that beta-carotene may affect cell growth by a redox mechanism. The purpose of this study was to determine whether the redox-sensitive transcription factor nuclear factor (NF)-kappaB may be involved in the growth inhibitory and proapoptotic effects of the carotenoid. To test this hypothesis, human leukemic cells (HL-60) and colon adenocarcinoma cells (LS-174 and WiDr) were treated with beta-carotene, alone or in combination with alpha-tocopherol or N-acetylcysteine, and changes in 1) cell oxidative status, 2) cell growth and apoptosis, 3) DNA-binding activity of NF-kappaB and 4) expression of c-myc, a NF kappaB target gene involved in apoptosis, were evaluated. In HL-60 cells, beta carotene induced a significant increase in reactive oxygen species (ROS) production (P < 0.001) and in oxidized glutathione (GSSG) content (P < 0.005) at concentrations >/=10 micro mol/L. These effects were always accompanied by a sustained elevation of NF-kappaB and by a significant inhibition (P < 0.002) of cell growth. NF-kappaB DNA-binding activity increased at 3 h and persisted for at least 48 h. Colon adenocarcinoma cells displayed substantial differences in their sensitivity to beta-carotene, exhibiting increased ROS levels and activation of NF-kappaB at concentrations much lower in LS-174 cells (2.5-5.0 micro mol/L) than in WiDr cells (50-100 micro mol/L). In all cell lines studied, alpha-tocopherol and N-acetylcysteine inhibited the effects of beta-carotene on NF-kappaB, cell growth and apoptosis, and normalized the increased expression of c-myc induced by the carotenoid. These data suggest that the redox regulation of NF-kappaB induced by beta-carotene is involved in the growth-inhibitory and proapoptotic effects of the carotenoid in tumor cells. PMID- 12566472 TI - Genistein and daidzein downregulate prostate androgen-regulated transcript-1 (PART-1) gene expression induced by dihydrotestosterone in human prostate LNCaP cancer cells. AB - Epidemiologic investigations and laboratory studies suggest that bioactive soy phytochemical components may be used as an effective dietary regimen for prevention of prostate cancer. Studies designed to identify new genes that are responsive to androgens and are sensitive to the prevention of prostate cancer using soy bioactive components have become a research priority. In this study, we determined the effect of soy isoflavones on the expression of prostate androgen regulated transcript 1 (PART-1), a newly discovered androgen-induced gene that may represent a novel androgen-dependent prostate cancer tumor marker. In an androgen-depleted cell culture system, 5alpha-dihydrotestosterone (DHT) induced expression of PART-1 transcript in androgen-sensitive LNCaP, but not in androgen independent DU 145 or PC-3 human prostate cancer cells. The soy isoflavones genistein and daidzein dose-dependently inhibited DHT-induced expression of the PART-1 transcript. Genistein at 50 micro mol/L completely inhibited expression of the PART-1 transcript in LNCaP cells induced by DHT at 0.1 and 1.0 nmol/L. Daidzein was less potent than genistein, whereas glycitein at the same levels as genistein or daidzein did not inhibit DHT-induced PART-1 transcript expression. Our studies suggest that use of the PART-1 gene as a biomarker for evaluating the efficacy of soy isoflavones on androgen-dependent prostate cancer warrants further investigation. PMID- 12566473 TI - Use of oral contraceptives blunts the calciuric effect of caffeine in young adult women. AB - Caffeine consumption increases the urinary excretion of calcium and other minerals. Factors that affect caffeine metabolism such as steroid hormones may modify this effect. The purpose of this study was to evaluate the influence of oral contraceptive (OC) use on the 4-h urinary excretion of calcium, phosphorus, magnesium, zinc, sodium, potassium and caffeine metabolites in response to a high caffeine dose given as coffee beverage. Adult women, 20-29 y, users (+OC, n = 15) and nonusers (-OC, n = 15) of oral contraceptives, with calcium intake approximately 500 mg/d, participated in two tests, caffeine load (5 mg/kg body weight) and no-caffeine control, in a randomized crossover design. The net increase (caffeine load corrected by no caffeine) in urinary excretion of most minerals was significantly higher in -OC than in +OC (P < 0.05), with the larger group difference for calcium (ninefold) followed by magnesium (twofold), zinc (onefold) and potassium (onefold). Net increases in urinary excretion of 1 methylurate and paraxanthine were about three- and fivefold higher, respectively, in -OC than in +OC (P < 0.05) whereas net increases in urinary excretion of 5 acetylamino-6-formylamino-3-methyluracil (AFMU) and 1,7-dimethylurate were over twofold higher in the +OC than in -OC (P < 0.05). Following the caffeine load, most urinary minerals showed negative correlation with urinary 1-methylurate in OC (R 15% above ideal body weight) were assigned to either a Protein Group [protein: 1.6 g/(kg. d); CHO <40% of energy] or CHO Group [protein: 0.8 g/(kg. d); CHO >55%]. Diets were equal in energy (7100 kJ/d) and fat (50 g/d). After 10 wk, the Protein Group lost 7.53 +/- 1.44 kg and the CHO Group lost 6.96 +/- 1.36 kg. Plasma amino acids, glucose and insulin were determined after a 12-h fast and 2 h after a 1.67 MJ test meal containing either 39 g CHO, 33 g protein and 13 g fat (Protein Group) or 57 g CHO, 12 g protein and 14 g fat (CHO Group). After 10 wk, subjects in the CHO Group had lower fasting (4.34 +/- 0.10 vs 4.89 +/- 0.11 mmol/L) and postprandial blood glucose (3.77 +/- 0.14 vs. 4.33 +/- 0.15 mmol/L) and an elevated insulin response to meals (207 +/- 21 vs. 75 +/- 18 pmol/L). This study demonstrates that consumption of a diet with increased protein and a reduced CHO/protein ratio stabilizes blood glucose during nonabsorptive periods and reduces the postprandial insulin response. PMID- 12566476 TI - A reduced ratio of dietary carbohydrate to protein improves body composition and blood lipid profiles during weight loss in adult women. AB - Claims about the merits or risks of carbohydrate (CHO) vs. protein for weight loss diets are extensive, yet the ideal ratio of dietary carbohydrate to protein for adult health and weight management remains unknown. This study examined the efficacy of two weight loss diets with modified CHO/protein ratios to change body composition and blood lipids in adult women. Women (n = 24; 45 to 56 y old) with body mass indices >26 kg/m(2) were assigned to either a CHO Group consuming a diet with a CHO/protein ratio of 3.5 (68 g protein/d) or a Protein Group with a ratio of 1.4 (125 g protein/d). Diets were isoenergetic, providing 7100 kJ/d, and similar amounts of fat ( approximately 50 g/d). After consuming the diets for 10 wk, the CHO Group lost 6.96 +/- 1.36 kg body weight and the Protein Group lost 7.53 +/- 1.44 kg. Weight loss in the Protein Group was partitioned to a significantly higher loss of fat/lean (6.3 +/- 1.2 g/g) compared with the CHO Group (3.8 +/- 0.9). Both groups had significant reductions in serum cholesterol ( approximately 10%), whereas the Protein Group also had significant reductions in triacylglycerols (TAG) (21%) and the ratio of TAG/HDL cholesterol (23%). Women in the CHO Group had higher insulin responses to meals and postprandial hypoglycemia, whereas women in the Protein Group reported greater satiety. This study demonstrates that increasing the proportion of protein to carbohydrate in the diet of adult women has positive effects on body composition, blood lipids, glucose homeostasis and satiety during weight loss. PMID- 12566477 TI - Selenium supplements do not increase plasma total homocysteine concentrations in men and women. AB - Studies in rats indicate that plasma total homocysteine (tHcy) is decreased in selenium deficiency and increases with selenium supplementation. The aim of this study was to determine the effect of selenium supplements on plasma tHcy concentrations in a population that has suboptimal selenium status. Subjects from Dunedin, New Zealand (n = 189) were randomly assigned to receive a supplement containing 200 micro g selenium or placebo for 20 wk. At baseline, 67% (n = 112) of the participants had plasma selenium concentrations < 1.2 micro mol/L, a concentration believed to be that necessary for full glutathione peroxidase (Gpx) activity. At 20 wk, plasma selenium concentration and Gpx activity increased in the selenium group by 1.2 micro mol/L [95% confidence interval (CI): 1.1, 1.3] and 5.1 nkat/g protein (3.7, 6.5), respectively, changes that were significantly greater than the changes in the placebo group (P < 0.001). At 20 wk, mean changes in plasma tHcy concentrations were 0.1 micro mol/L (95% CI: -0.4, 0.5) and -0.2 micro mol/L (-0.7, 0.3) in the supplemented and placebo groups, respectively, compared to baseline. There was no difference in the mean changes in plasma tHcy between the supplemented and placebo groups (P = 0.54). These results suggest that selenium supplementation does not influence plasma tHcy concentrations in healthy populations in developed countries, whose selenium status is characterized by lower plasma selenium concentrations. PMID- 12566478 TI - An evaluation of the U.S. Department of Agriculture food security measure with generalized linear mixed models. AB - Over the last decade, new information has been developed and collected to measure the extent of food insecurity and hunger in the United States. Common measurement of the phenomenon of hunger and food insecurity has become possible through efforts of the U.S. Department of Agriculture (USDA) to develop a set of survey questions that can be used to obtain estimates of the prevalence and severity of food insecurity. We evaluated the measurement of food insecurity and the effect of household variables on measured food insecurity. The effects of demographic and survey-specific variables on the food insecurity/hunger scale were evaluated using a generalized linear model with mixed effects. Data came from the 1995, 1997 and 1999 Food Security Module of the Current Population Survey. The results generally validated the model currently used by the USDA. In addition, our approach made it possible to consider the effect of demographics and several survey design variables on food security among measurably food-insecure households, as well as interactions between these factors and the food security questions. The analysis of the expanded model with the 1995 data found results similar to those reported based on the Rasch model used by the USDA. Even though the sample size was reduced and a number of screening and questionnaire changes were introduced in 1997 and 1999, the results for those years appear mostly unchanged and confirm the robustness of the scale in measuring food insecurity. There is some evidence that interpretation of questions may vary among different demographic groups. PMID- 12566479 TI - Dietary alpha-tocopherol decreases alpha-tocotrienol but not gamma-tocotrienol concentration in rats. AB - We previously showed that alpha- and gamma-tocotrienols accumulate in adipose tissue and skin but not in plasma or other tissues of rats fed a tocotrienol-rich fraction extracted from palm oil containing alpha-tocopherol and alpha- and gamma tocotrienols. To clarify the nature of tocotrienol metabolism, we studied the distribution of alpha- or gamma-tocotrienol in rats fed alpha- or gamma tocotrienol without alpha-tocopherol, and the effect of alpha-tocopherol on their distribution. Wistar rats (4-wk-old) were fed a diet with 50 mg alpha tocotrienol/kg alone or with 50 mg alpha-tocopherol/kg in expt. 1, and a diet with 50 mg gamma-tocotrienol/kg alone or with 50 mg alpha-tocopherol/kg in expt. 2, for 8 wk. alpha-Tocotrienol was detected in various tissues and plasma of the rats fed alpha-tocotrienol alone, and the alpha-tocotrienol concentrations in those tissues and plasma decreased (P < 0.05) by the dietary alpha-tocopherol in the rats fed alpha-tocotrienol with alpha-tocopherol. However, gamma-tocotrienol preferentially accumulated in the adipose tissue and skin of the rats fed gamma tocotrienol alone, and the dietary alpha-tocopherol failed either to decrease (P >/= 0.05) gamma-tocotrienol concentrations in the adipose tissue and skin or to increase (P >/= 0.05) in the urinary excretion of 2,7,8-trimethyl-2(2' carboxymethyl)-6-hydroxycroman, a metabolite of gamma-tocotrienol, in the rats fed gamma-tocotrienol with alpha-tocopherol. These data suggest that alpha tocopherol enhances the alpha-tocotrienol metabolism but not the gamma tocotrienol metabolism in rats. PMID- 12566480 TI - Stereospecific incorporation of palmitoyl, oleoyl and linoleoyl moieties into adipose tissue triacylglycerols of rats results in constant sn-1:sn-2:sn-3 in rats fed rapeseed, olive, conventional or high oleic sunflower oils, but not in those fed coriander oil. AB - We report the stereospecific (sn-1, sn-2, sn-3) distribution of fatty acids in subcutaneous adipose tissue triacylglycerols of male weaned Wistar rats fed either a standard diet or diets containing, in addition to 20 g corn oil/kg feed, 120 g/kg feed, each, of canola-type rapeseed oil, olive oil, conventional or high oleic sunflower oil or high petroselinic coriander oil for 10 wk. The regiospecific distribution of the major acyl moieties in the sn-1 (3) vs. sn-2 positions of the adipose tissue triacylglycerols broadly reflected that of the dietary oils. The saturated palmitoyl and stearoyl moieties were more abundant in the sn-1 and sn-3 positions compared with the sn-2 position of the adipose tissue triacylglycerols, and both occurred at a higher proportion in the sn-1 than in the sn-3 position. Oleoyl moieties were abundant in all the three positions of the adipose tissue triacylglycerols, whereas petroselinoyl moieties were more abundant in the sn-1 and sn-3 positions compared with the sn-2 position. Linoleoyl moieties occurred predominantly in the sn-2 position compared with the sn-1 and sn-3 positions of the adipose tissue triacylglycerols; however, they were more abundant in the sn-3 than in the sn-1 position. Despite widely varying proportions of the palmitoyl, oleoyl and linoleoyl moieties at the three positions of the dietary triacylglycerols, the ratios of each of these acyl moieties at the sn-1, sn-2, and sn-3 positions in adipose tissue triacylglycerols were essentially constant for all groups, with the exception of the group fed coriander oil, indicating a rigid stereospecific incorporation. PMID- 12566481 TI - Inadequate copper intake reduces serum insulin-like growth factor-I and bone strength in growing rats fed graded amounts of copper and zinc. AB - This study examined the effects of graded intakes of zinc (Zn) and copper (Cu) on serum insulin-like growth-factor-I (IGF-I) concentration and bone quality in growing rats. Using a 3 x 4 factorial design, weanling male Sprague-Dawley rats were randomly assigned to 12 groups (n = 7 per group) and were fed one of nine modified AIN-93G basal diets with varying amounts of Cu (0.3, 3 and 10 micro g/g) and Zn (5, 15 and 45 micro g/g) for 6 wk. A group of rats was pair-fed to each low Zn group. Although dietary Zn mainly influenced body weights (P < 0.0001), dietary Cu was the main determinant of most of the variables related to bone quality. Low Cu intake reduced serum IGF-I and femur breaking force and ultimate stress (by 27, 14 and 7%, respectively; P < 0.05) and increased bone IGF-I concentration (by 62%; P < 0.0001). Low Cu intake also increased femur nitrogen, hydroxyproline, hexosamine and calcium (Ca) concentrations of long bones (P < 0.05). Lumbar vertebrae dry weight and density were the lowest in the rats fed the low Cu diets (P < 0.001) and were higher in the rats fed high amounts of both Cu and Zn (P < 0.01). In summary, growing rats fed low and marginal Cu had lower serum IGF-I than those fed high dietary Cu. Bone strength was also reduced with low dietary Cu, despite compensatory changes in the bone matrix. In the presence of graded intakes of Cu, the effects of low dietary Zn were more pronounced on the spinal bones than the long bones. PMID- 12566482 TI - Breast milk fractions solubilize Fe(III) and enhance iron flux across Caco-2 cells. AB - Why breastfed infants absorb extrinsic iron (EFe) exceptionally well is an unexplained phenomenon. Our objective was to identify effects of human milk fractions (HMF) on bioavailability of EFe. HMF were prepared by centrifugation followed by successive ultrafiltration using 10-, 3- and 1-kDa molecular weight cutoff membranes. EFe was added to HMF before and after treatment with digestive enzymes. Solubilization of EFe by HMF was characterized by scintillation counting of radioiron and by size exclusion chromatography/inductively coupled plasma mass spectrometry (SEC/ICPMS) of stable iron. Effects of HMF on EFe uptake and basolateral transfer were assessed by using confluent Caco-2 cells in bicameral chambers. Whey fractions of low molecular weight (MW) derived from 10-kDa filtrate, except the 1-kDa filtrate, were as effective as ascorbate and nitrilotriacetate in solubilizing EFe at intestinal pH. Basolateral radioiron transfer from Caco-2 cell monolayers was greater in the presence of low MW whey fractions than in the presence of ferrous ascorbate. The 3-kDa filtrate and 3-kDa retentate fractions promoted basolateral transfer of cellular radioiron taken up previously. SEC/ICPMS of the 1-kDa retentate fraction revealed a UV-absorbing peak of MW approximately 4.2 kDa that contained iron and that solubilized added ferric iron both before and after in vitro digestion with pepsin, pancreatin and bile extract. Our results suggested that a low MW component of breast milk whey enhances iron bioavailability. Because the iron solubilization activity is resistant to in vitro digestion, it is plausible that the component is active in vivo and may explain the excellent absorption of EFe by breastfed infants [corrected]. PMID- 12566483 TI - Dietary conjugated linoleic acid lowers plasma cholesterol during cholesterol supplementation, but accentuates the atherogenic lipid profile during the acute phase response in hamsters. AB - Conjugated linoleic acid (CLA) reportedly exerts anticarcinogenic and antiatherosclerotic effects in animals. To test the hypothesis that the putative antiatherosclerotic effect of CLA might derive from an anti-inflammatory or antioxidant action on lipoprotein metabolism, an acute phase response (APR) was elicited in hamsters while varying dietary cholesterol and vitamin E intakes in two experiments. The effect of CLA intake (to 1%) was examined with 0% (Experiment 1, 7 wk) and 0 or 0.3% (Experiment 2, 12 wk) cholesterol, at which point APR was induced. In hamsters not fed dietary cholesterol (Experiment 1), CLA exaggerated the rise in plasma and LDL cholesterol observed during the APR. When CLA was fed concurrently with cholesterol (Experiment 2), plasma and liver cholesterol were reduced up to 40% independent of the APR. In addition, CLA decreased body weight gain and adipose reserves in Experiment 1, but not in Experiment 2. Because CLA failed to attenuate APR and was not influenced by vitamin E status, an antioxidant/anti-inflammatory role was not apparent. However, the reduced burden on liver and lipoprotein cholesterol induced by CLA during cholesterol feeding, suggests that CLA curtailed cholesterol absorption, whereas the rise during APR suggests that CLA exaggerated the impaired clearance of plasma cholesterol associated with acute inflammation. PMID- 12566484 TI - Microbial aromatic acid metabolites formed in the gut account for a major fraction of the polyphenols excreted in urine of rats fed red wine polyphenols. AB - The health effects of dietary polyphenols might be explained by both intact compounds and their metabolites formed either in the tissues or in the colon by the microflora. The quantitative importance and biological activities of the microbial metabolites have seldom been examined in vivo. We measured the microbial metabolites formed in four groups of rats (n = 8) fed for 8 d a diet supplemented with 0.12 g/100 g catechin, 0.25 or 0.50 g/100 g red wine powder containing proanthocyanidins, phenolic acids, flavanols, anthocyanins and flavonols or an unsupplemented diet. Fourteen aromatic acid metabolites were assayed in urine collected for 24 h by an HPLC-electrospray ionization (ESI)-mass spectrometry (MS)-MS method. The three main metabolites formed from the catechin diet were 3-hydroxyphenylpropionic acid, 3-hydroxybenzoic acid and 3 hydroxyhippuric acid. Their total urinary excretion accounted for 4.7 g/100 g of the catechin ingested and that of intact catechins for 45.3 g/100 g. For wine polyphenols, the same microbial metabolites as observed for the catechin diet were identified in urine along with hippuric, p-coumaric, vanillic, 4 hydroxybenzoic and 3-hydroxyphenylacetic acids. All together, these aromatic acids accounted for 9.2 g/100 g of the total wine polyphenols ingested and intact catechins for only 1.2 g/100 g. The higher excretion of aromatic acids by rats fed wine polyphenols is likely due to their poor absorption in the proximal part of the gut. Some of the microbial metabolites still bear a reducing phenolic group and should also prevent oxidative stress in inner tissues. More attention should be given in the future to these microbial metabolites and their biological properties to help explain the health effects of polyphenols that are not easily absorbed through the gut barrier. PMID- 12566485 TI - Beta-glucan fractions from barley and oats are similarly antiatherogenic in hypercholesterolemic Syrian golden hamsters. AB - The cholesterol-lowering activities of oats and barley are commonly attributed to the beta-glucan fractions. Although beta-glucan is present in both grains and appears to be chemically similar, the effect of source on cholesterol-lowering activity has not been evaluated. In the present study, the antiatherogenic properties of beta-glucan concentrates from oats and barley were evaluated in Syrian golden F(1)B hamsters consuming a semipurified hypercholesterolemic diet (HCD) containing cholesterol (0.15 g/100 g), hydrogenated coconut oil (20 g/100 g) and cellulose (15 g/100 g). After a 2-wk lead-in period, control hamsters were fed the HCD, whereas experimental hamsters consumed HCD formulated to include beta-glucan (2, 4, or 8 g/100 g) by addition of beta-glucan concentrate prepared from oats or barley at the expense of cellulose. Compared with control hamsters, dose-dependent decreases that were similar in magnitude in plasma total and LDL cholesterol concentrations were observed in hamsters fed beta-glucan from either source at wk 3, 6 and 9. Compared with controls, liver cholesterol concentrations were also reduced (P < 0.05) in hamsters consuming 8 g/100 g oat or barley beta glucan. In agreement with previously proposed mechanisms, total fecal neutral sterol concentrations were significantly increased (P < 0.05) in hamsters consuming 8 g/100 g barley or oat beta-glucan. Aortic cholesterol ester concentrations were significantly reduced (P < 0.05) in hamsters fed 8 g/100 g beta-glucan from barley or oats. Although aortic total cholesterol and cholesterol ester concentrations were significantly correlated with LDL cholesterol (r = 0.565, P < 0.004 and r = 0.706, P < 0.0001, respectively), this association could explain only half of the variability. This study demonstrated that the cholesterol-lowering potency of beta-glucan is approximately identical whether its origin was oats or barley. PMID- 12566486 TI - Neither methionine nor nitrous oxide inactivation of methionine synthase affect the concentration of 5,10-methylenetetrahydrofolate in rat liver. AB - 5,10-Methylenetetrahydrofolate occupies a key position in folate-dependent one carbon metabolism. It is involved directly in the biosynthesis of deoxythymidine, it can be converted to 10-formyltetrahydrofolate for purine synthesis and it may be reduced to 5-methyltetrahydrofolate for methylation of homocysteine to methionine. We have developed a HPLC method for measuring 5,10 methylenetetrahydrofolate in liver and we have used this method to investigate two conditions that perturb one-carbon metabolism: 1) administration of methionine and 2) administration of the anesthetic gas, nitrous oxide (N(2)O). Rats were given 1.3 mmol/kg of methionine, and folate coenzymes in liver were measured. As expected, giving methionine resulted in an apparent increase in the concentration of 10-formyl- and tetrahydrofolate and an apparent decrease in 5 methyltetrahydrofolate concentration at 30 and 60 min. After 120 min, the concentrations of these coenzymes appeared to revert to control values. There was no apparent change in the concentration of 5,10-methylenetetrahydrofolate. Exposing rats to an atmosphere containing N(2)O results in inactivation of methionine synthase and accumulation of 5-methyltetrahydrofolate at the expense of other folate coenzymes. In liver from rats breathing N(2)O, 5 methyltetrahydrofolate increased, whereas there was no change in 5- or 10 formyltetrahydrofolates (P > 0.7 and P > 0.8, respectively). Tetrahydrofolate was not detected in liver from the N(2)O group, whereas it constituted 24% of folates in the control group. The concentration of 5,10-methylenetetrahydrofolate was not significantly affected by N(2)O (P > 0.18). These results suggest that the concentration of 5,10-methylenetetrahydrofolate is tightly regulated in liver. PMID- 12566487 TI - Tissue antioxidant status differs in spontaneously hypertensive rats fed fish protein or casein. AB - The present study was designed to determine whether changes in dietary protein source are related to changes in antioxidant status determined by enzyme activities of catalase, superoxide dismutase (SOD), gluthatione peroxidase (GSH Px) and gluthatione reductase (GSSG-Red) and lipid peroxidation levels in various tissues. Spontaneously hypertensive rats (SHR; 5 wk old) were fed diets containing 20% casein or fish protein for 2 mo. Feeding the fish protein diet lowered blood pressure and reduced plasma total cholesterol levels and SOD activity in all tissues except muscle compared with the casein diet. Feeding fish protein also enhanced GSH level and GSH-Px activity in liver and heart, accompanied by lower lipid peroxidation. In kidney, however, the lower catalase activity in rats fed fish protein was associated with an enhancement in lipid peroxidation. Plasma and VLDL + LDL lipid peroxidation was unaffected by dietary proteins. In conclusion, the fish protein diet did not play a relevant role in plasma antioxidative defense status but increased it in liver and heart compared with the casein diet. Fish protein attenuated the development of hypertension and also decreased plasma total cholesterol concentration. Thus, it enhances protection against cardiovascular diseases. PMID- 12566488 TI - Ferric citrate is half as effective as ferrous sulfate in meeting the iron requirement of juvenile tilapia, Oreochromis niloticus x O. aureus. AB - Two growth experiments were conducted to estimate the minimum dietary iron requirement for juvenile hybrid tilapia, Oreochromis niloticus x O. aureus. Purified diets containing 0, 10, 30, 50, 100, 150, 200 and 400 mg Fe/kg from ferric citrate (Experiment 1) and 0, 10, 30, 50, 100, 150 and 200 mg Fe/kg from ferrous sulfate (Experiment 2) were fed to tilapia (mean initial weight: 0.63 +/- 0.01 g, Experiment 1; 0.64 +/- 0.01 g, Experiment 2) for 8 wk. In Experiment 2, 150 mg Fe/kg from ferric citrate was also included for comparison. The rearing water contained 1.07 micro mol/L iron, and supplemental levels were confirmed by analysis. Each diet was fed to three replicate groups of fish. In Experiment 1, weight gain and feed efficiency (FE) were highest (P < 0.05) in fish fed the diet supplemented with 150 mg Fe/kg, followed by fish fed diets with 50, 100 and 200 mg Fe/kg and lowest in fish fed the unsupplemented control diet. Hepatic iron concentration was highest in fish fed diets supplemented with >150 mg Fe/kg, followed by fish fed the diet with 100 mg Fe/kg and lowest in fish fed diets with /=100 mg Fe/kg and mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) were higher in fish fed diets with >/=150 mg Fe/kg than in fish fed the diet without iron supplementation. In Experiment 2, weight gain was higher in fish fed the diet with 50 mg Fe/kg than in fish fed diets with 150, 200 and /=50 mg Fe/kg and the ferric citrate comparison diet than fish fed diets with /=50 mg Fe/kg than in fish fed the unsupplemented control diet. Analyses by polynomial regression of weight gain and by broken-line regression of hepatic iron and blood Hb concentrations indicated that the dietary iron requirement for tilapia is approximately 150-160 mg Fe/kg and 85 mg Fe/kg with ferric citrate and ferrous sulfate as the iron source, respectively; it also appears that ferric citrate was approximately 50% as effective as ferrous sulfate in meeting the iron requirement. PMID- 12566489 TI - Dietary interactions influence the effects of bovine folate-binding protein on the bioavailability of tetrahydrofolates in rats. AB - The newborns of mammals have a high folate demand, yet obtain adequate folate nutrition solely from their mothers' milk despite its low folate content. Milk folate is entirely bound by an excess of folate-binding protein (FBP), prompting speculation that FBP may affect the bioavailability of the limited folate supply. Previous research has shown that FBP-bound folic acid is more gradually absorbed, thereby reducing the peak plasma folate concentration and preventing loss into the urine. Natural folates are reduced derivatives of folic acid, with milk predominantly containing 5-methyltetrahydrofolate, yet little research has been carried out to determine the role of FBP in the bioavailability of reduced folates. We studied the effect of FBP on folate nutrition of rats in both single dose and 4-wk feeding experiments. The effect of FBP was influenced by the presence of other milk components. FBP increased bioavailability of dietary folate when it was consumed with other whey proteins or with soluble casein. However, in the presence of acid-precipitated casein or a whey preparation enriched in lipids, bioavailability was decreased. These results highlight the difficulties of extrapolating from experimental results obtained using purified diets alone and of studying interactions among dietary components. They suggest that the addition of FBP-rich foods to folate-rich foods could enhance the bioavailability of natural folates, but that the outcome of such a combination would depend on interactions with other components of the diet. PMID- 12566490 TI - (n-3) Polyunsaturated fatty acids promote activation-induced cell death in murine T lymphocytes. AB - Previous studies showing dietary (n-3) polyunsaturated fatty acids (PUFA) attenuate T cell immune-mediated inflammatory diseases led us to hypothesize that (n-3) PUFA promote activation-induced cell death (AICD) in T cells. Because T cell subsets display a differential resistance to AICD, we compared the effects of (n-3) PUFA feeding on T cells stimulated in vitro to express different cytokine profiles. Mice were fed either diets lacking (n-3) PUFA (control) or (n 3) PUFA-containing diets for 14 d. Splenic T cells were stimulated with alphaCD3/alphaCD28, phorbol myristate acetate (PMA)/Ionomycin or alphaCD3/PMA for 48 h, followed by reactivation with the same stimuli for 5 h. Apoptosis was measured using Annexin V/propidium iodide. (n-3) PUFA were selectively incorporated into membrane phospholipid pools. Cytokine analyses revealed that (n 3) PUFA enhanced AICD only in T cells expressing a T helper cell (Th)1-like cytokine profile after stimulation with PMA/Ionomycin compared to mice fed the (n 6) PUFA control diet (P = 0.0008). In contrast, no increase in apoptosis was seen in T cells stimulated with alphaCD3/PMA, which exhibited a Th2 cytokine profile. These data demonstrate that the ability of (n-3) PUFA to promote AICD is dependent on the activation stimulus. In conclusion, we have identified a novel mechanism by which (n-3) PUFA modulate T cell-mediated immunity by selective deletion of Th1-like cells while maintaining or enhancing the Th2-mediated humoral immune response. PMID- 12566491 TI - Leptin suppresses food intake and body weight in corticosterone-replaced adrenalectomized rats. AB - Intracerebroventricular (ICV) injections of leptin decrease food intake and body weight while increasing energy expenditure. Some of these effects are reportedly enhanced in bilaterally adrenalectomized (ADX) rats. The purpose of the present experiment was to establish the time course of the suppression in body weight and food intake after an ICV injection of leptin. We wanted to establish the effect of varying doses of corticosterone (CORT) on body weight and food intake suppression by using separate groups of ADX, ADX and corticosterone-treated and sham-operated Sprague-Dawley rats. All rats were implanted with cholesterol pellets that varied in CORT content. During the same surgical session, all rats were fitted with a cannula in the lateral ventricle. After recovering from surgery, each rat was administered a 5- micro g ICV injection of leptin. ADX rats that were treated with CORT replacement lost more (P < 0.05) weight and took longer (P < 0.05) to return to baseline body weight than sham-operated controls. Leptin injection decreased food consumption to a greater extent (P < 0.05) in the ADX groups treated with CORT than in the sham-operated controls. Plasma insulin increased in a dose-dependent manner in the ADX rats as a function of CORT replacement. The higher of the two CORT replacement doses used in this experiment restored circulating CORT to levels observed in sham-operated controls. Contrary to earlier reports, physiological doses of CORT appear to enhance leptin-induced weight loss. PMID- 12566492 TI - Portion size adds limited information on variance in food intake of participants in the EPIC-Potsdam study. AB - Food-frequency questionnaire (FFQ) data should reflect interindividual variation and therefore measure variance in intake among populations. We conducted this analysis to evaluate the relevance of separate portion size questions to the interindividual variation in food intake. The contribution of portion size questions to the variance in food intake was quantified and compared with the variance when group-specific portion sizes would be assigned, using 26,764 FFQ of the European Investigation into Cancer and Nutrition (EPIC)-Potsdam Study. Groups were defined according to gender, age (<50 y, >/=50 y) or body mass index (BMI) (<26 kg/m(2), >/=26 kg/m(2)). The FFQ inquired about both consumption frequency and portion size. Linear regression models for each food item were fit with intake (g/d) as dependent variables and frequency of intake as independent variables. The mean coefficient of determination (R(2)) for the different food items explained by frequency only was 84.0% (71.2-95.7%). The R(2) for gender-, age- and BMI-specific frequencies of intake did not markedly alter the overall results. We conclude that the omission of individual portion size information would probably result in a notable reduction of interindividual variance. However, to reduce the respondents' burden and to increase data completeness in self-administration in large epidemiologic studies, the assignment of a constant portion size seems to be adequate. The variance was not increased markedly when constant gender-, age- and BMI-specific portion sizes were applied, thus supporting the assignment of an overall portion size. PMID- 12566493 TI - Soy phytochemicals and tea bioactive components synergistically inhibit androgen sensitive human prostate tumors in mice. AB - Although high doses of single bioactive agents may have potent anticancer effects, the chemopreventive properties of the Asian diet may result from interactions among several components that potentiate the activities of any single constituent. In Asia, where intake of soy products and tea consumption are very high, aggressive prostate cancer is significantly less prevalent in Asian men. The objective of the present study was to identify possible synergistic effects between soy and tea components on prostate tumor progression in a mouse model of orthotopic androgen-sensitive human prostate cancer. Soy phytochemical concentrate (SPC), black tea and green tea were compared with respect to tumorigenicity rate, primary tumor growth, tumor proliferation index and microvessel density, serum androgen level and metastases to lymph nodes. SPC, black tea and green tea significantly reduced tumorigenicity. SPC and black tea also significantly reduced final tumor weights. Green tea did not reduce final tumor weight, although it tended to elevate (P = 0.14) the serum dihydrotestosterone (DHT) concentration. The combination of SPC and black tea synergistically inhibited prostate tumorigenicity, final tumor weight and metastases to lymph nodes in vivo. The combination of SPC and green tea synergistically inhibited final tumor weight and metastasis and significantly reduced serum concentrations of both testosterone and DHT in vivo. Inhibition of tumor progression was associated with reduced tumor cell proliferation and tumor angiogenesis. This study suggests that further research is warranted to study the role of soy and tea combination as effective nutritional regimens in prostate cancer prevention. PMID- 12566494 TI - Low dietary copper increases fecal free radical production, fecal water alkaline phosphatase activity and cytotoxicity in healthy men. AB - One possible dietary factor that may increase susceptibility to colon cancer is inadequate copper intake. The objective of this study was to investigate the effects of low and adequate copper intakes on copper nutriture and putative risk factors for colon cancer susceptibility in healthy men. Seventeen healthy free living nonsmoking men aged 21-52 y completed a 13-wk controlled feeding study in a randomized crossover design. The basal diet contained 0.59 mg Cu/13.65 MJ. After a 1-wk equilibration period in which the men consumed the basal diet supplemented with 1.0 mg Cu/d, they were randomly assigned to receive either the basal diet or the basal diet supplemented with 2 mg Cu/d for 6 wk. After the first dietary period, the men immediately began to consume the other level of Cu for the last 6 wk. They collected their feces during the equilibration period and during the last 2 wk of the two dietary periods for free radical and fecal water analysis. Low dietary copper significantly (P < 0.01) increased fecal free radical production and fecal water alkaline phosphatase activity. Low dietary copper significantly (P < 0.0001) decreased fecal water copper concentrations but did not affect fecal water volume, pH, iron or zinc concentrations. In contrast to the fecal analysis, hematological indicators of copper status were not significantly affected by the dietary treatments. These results suggest that low dietary copper adversely affects fecal free radical production and fecal water alkaline phosphatase activity, which are putative risk factors for colon cancer. PMID- 12566495 TI - Vitamin E supplementation does not alter azoxymethane-induced colonic aberrant crypt foci formation in young or old mice. AB - Vitamin E, part of the body's primary lipid-soluble defense against free radicals and reactive oxygen molecules, has been suggested to reduce the risk for some cancers. However, the role of vitamin E in the etiology and prevention of colon cancer, especially in the highest risk group, the aged, is not clear. Thus, this study was conducted to elucidate the effect of vitamin E supplementation on susceptibility to colon cancer by examining azoxymethane (AOM)-induced aberrant crypt foci (ACF) formation, a surrogate biomarker of colon cancer. Young (3-4 mo) and old (19-20 mo) C57BL/6JNIA mice were fed either a control diet (30 mg dl alpha-tocopheryl acetate/kg diet) or a vitamin E-supplemented diet (500 mg dl alpha-tocopheryl acetate/kg diet) for 16 wk. After 6 wk of dietary supplementation, young and old mice were injected with saline or AOM weekly for 5 wk to receive the same total dose of AOM (2.2 mg) and killed 10 wk after the first AOM injection. Vitamin E supplementation had no effect on the number of AOM induced ACF in young or old mice. In addition, vitamin E supplementation did not have an effect on splenocyte interferon-gamma, interluekin-6 and tumor necrosis factor-alpha levels, natural killer cell killing activity or colonic cell proliferation in young or old mice. Thus, alpha-tocopherol does not seem to affect the initiation and early promotion stages of AOM-induced colon carcinogenesis in young or old mice. Whether vitamin E supplementation might be effective in reducing AOM-induced colon tumors is unclear. PMID- 12566496 TI - Comment on Dickinson et al. (2002). PMID- 12566498 TI - Innovative teaching strategies for training physicians in clinical nutrition: an overview. PMID- 12566499 TI - Residents as teachers: a process for training and development. AB - Of the many roles that residents fill, an important one is that of teacher. Residents spend many hours teaching a variety of learners-peers, patients, medical students, the general public-in a range of environments, from the bedside and clinic, to the OR, conference room and lecture hall. Such diverse audiences and surroundings almost command the use of a wide array of teaching strategies appropriate both to the setting and to the learner. This paper addresses how residents can learn to be effective teachers without taxing an already-overloaded schedule. This focus is on the process of developing a learning format that is appropriate to any given discipline by means of a partnership among an educator, a faculty member and residents from within a discipline. A brief review of the educational literature provides the basis on which are built actual teaching strategies for teaching residents to be teachers. PMID- 12566500 TI - Pocket tape measure for waist circumference: training medical students and residents on a simple assessment of body composition. AB - Metabolic syndrome, the clustering of hyperglycemia, hypertension and dyslipidemia, increases the risk of coronary heart disease. Abdominal obesity is an important cue for the clinician to consider metabolic syndrome. Measurement of waist circumference is a simple means of identifying abdominal obesity. The development and distribution of pocket tape measures to medical students, residents and attending physicians were initiated to enhance identification and treatment of metabolic syndrome. Distribution of the tape measures was added to a cardiovascular nutrition component in a 4th-y medical school curriculum. The nutrition component continued to include computer-based cases and pocket reference cards. Limited data suggest that the addition of pocket tape measures to the nutrition component of an ambulatory care clerkship may increase the percentage of medical students who use waist circumference to identify patients at risk for metabolic syndrome. It is anticipated that student use will increase with role modeling by residents and attending physicians. PMID- 12566501 TI - Cardiovascular risk factor self-assessment program: using the general clinical research center to provide a clinical experience for third-year medical students. AB - An educational program at the University of Iowa has been designed to provide medical students with the opportunity to do a self-assessment of their personal risk factors for developing cardiovascular disease. The University of Iowa Hospital and Clinic's General Clinical Research Center (GCRC) provided the personnel and resources to work with students in small groups to allow them to experience the diagnostic testing that is a standard part of cardiovascular assessment procedures. This report presents preliminary data from the first 88 students to participate in the program. PMID- 12566502 TI - REAP and WAVE: new tools to rapidly assess/discuss nutrition with patients. AB - Dietary changes can be helpful in preventing or treating a variety of prevalent health problems. Physicians can be helpful in helping patients make positive dietary changes, be physically active and lose weight, but, for a variety of reasons, many physicians do little nutrition counseling. There is a need for brief, user-friendly tools to enable physicians to rapidly and accurately assess patients' diets and exercise habits as well as provide information to aid the physician in delivering effective nutrition counseling. The purpose of this paper is to discuss two new tools, WAVE and REAP, that have been developed by the Nutrition Academic Award to help physicians and other health care providers conduct nutrition assessment and counseling with their patients in a practical and effective manner. The WAVE acronym and tool is designed to encourage provider/patient dialogue about the pros and cons of the patients' current status related to Weight, Activity, Variety and Excess. The Rapid Eating and Activity Assessment for Patients (REAP) is a brief validated questionnaire that is designed to aid providers in performing a brief assessment of diet and physical activity. An accompanying Physician Key aids the provider in discussing the patient's answers and counseling them appropriately. REAP and WAVE can be helpful tools to facilitate nutrition assessment and counseling in the provider office. Depending on patients' health priorities and how much time is available, these tools can be used in a variety of ways to discuss nutrition with patients during a clinical encounter in 1-9 min. PMID- 12566503 TI - Teaching nutrition skills to primary care practitioners. AB - Primary care physicians have the potential to decrease morbidity and mortality for many chronic diseases if they provide effective nutrition counseling. Given the time constraints of primary care practice, nutrition counseling needs to be brief, be part of an organized office system and refer appropriate patients to qualified nutrition professionals to be effective. This paper reviews a system of primary care nutrition counseling using the 5A's of patient-centered counseling, the elements necessary to develop an office-based system and some successful tools developed by nutrition researchers for the primary care setting to be used in an office-based system. PMID- 12566504 TI - Longitudinal and horizontal integration of nutrition science into medical school curricula. AB - The overall goal of our Nutrition Academic Award (NAA) medical nutrition program at Mercer University School of Medicine is to develop, implement and evaluate a medical education curriculum in nutrition and other aspects of cardiovascular disease (CVD) prevention and patient management with emphasis on the training of primary care physicians for medically underserved populations. The curriculum is 1) vertically integrated throughout all 4 y of undergraduate medical education, including basic science, clinical skills, community science and clinical clerkships as well as residency training; 2) horizontally integrated to include allied healthcare training in dietetics, nursing, exercise physiology and public health; and 3) designed as transportable modules adaptable to the curricula of other medical schools. The specific aims of our program are 1) to enhance our existing basic science problem-based Biomedical Problems Program with respect to CVD prevention through development of additional curriculum in nutrition/diet/exercise and at-risk subpopulations; 2) to integrate into our Clinical Skills Program objectives for medical history taking, conducting patient exams, diet/lifestyle counseling and referrals to appropriate allied healthcare professionals that are specific to CVD prevention; 3) to enhance CVD components in the Community Science population-based medicine curriculum, stressing the health-field concept model, community needs assessment, evidence-based medicine and primary care issues in rural and medically underserved populations; 4) to enhance the CVD prevention and patient management component in existing 3rd- and 4th-y clinical clerkships with respect to nutrition/diet/exercise and socioeconomic issues, behavior modification and networking with allied health professionals; and 5) to integrate a nutrition/behavior change component into Graduate Residency Training in CVD prevention. PMID- 12566506 TI - Future directions for the integrated CSFII-NHANES: What We Eat in America-NHANES. AB - The history of the integration of the dietary data collection from the National Health and Nutrition Examination Survey (NHANES) and the Continuing Survey of Food Intakes by Individuals (CSFII) is reviewed. The purposes and process of the workshop are presented. The three key topics of the workshop are summarized. The key roles of cosponsors and participants are acknowledged. PMID- 12566507 TI - Integrated NHANES: uses in national policy. AB - The National Nutrition Monitoring and Related Research Program (NNMRRP) arose from Congressional concern about lack of information regarding the occurrence in the American population of undernutrition and diet-related risk factors for chronic diseases. Congressional appropriations and executive branch decisions about budget priorities have been the major determinants of the scope and number of nutrition monitoring surveys and surveillance activities fielded and therefore the information available for policy and research uses. The nutrition data collected in the NNMRRP are used by federal agencies, the private sector and academia for a variety of purposes, including public policy (e.g., development and evaluation of monitoring and surveillance, regulatory and nutrition programs), normative standards (e.g., growth charts, reference data for hematological and biochemical indicators of nutritional status or Dietary Reference Intakes) and research (e.g., cross-sectional, longitudinal and time trends studies of dietary and nutritional status, health status, disease morbidity and mortality). Although the importance of the NNMRRP to national policy is difficult to quantify, in a 5-y period 97 proposed and final regulations citing NNMRRP data were published in the Federal Register by federal agencies responsible for nutrition and food safety programs. The NNMRRP-derived dietary and nutritional status data are essential information for quantitative risk assessments increasingly relied on by regulatory agencies as the basis for programmatic decisions and regulations development. Users of NNMRRP data in government agencies, academic institutions and the private sector have come to recognize the value of data from the surveys and surveillance systems for a wide variety of programmatic and research purposes. PMID- 12566508 TI - Collection and analysis of intake data from the integrated survey. AB - Intake data from the combined CSFII/NHANES survey will be used for many different purposes, each with specific data requirements and appropriate analytic methods. For monitoring and surveillance, the availability of Dietary Reference Intakes will allow estimates of the prevalence of inadequate intakes and the prevalence of intakes with a risk of adverse effects. The accuracy of the nutrient intake estimates will be enhanced by the 5-pass dietary recall methodology, availability of quantified dietary supplement intake data and expanded food and supplement composition data. Food-level dietary monitoring will be improved by using new databases to calculate servings of food groups from the Food Guide Pyramid and intakes of food commodities. Another major strength of the survey is the ability to relate intake data to health measures for individuals. Inferences will continue to be limited by a lack of usual intake for each individual, but the attenuation will be less with 2 d of data than with only 1 d, as in the past. Better data collection and analysis will also lead to more informed nutrition policies and programs. Innovative methods of analyzing the data should be investigated to minimize the effects of underreporting, provide better estimates of usual intake at both the group and individual levels and accurately combine nutrient intakes from foods and supplements. Future modifications to the intake collection methods might be considered to allow larger sample sizes for certain subgroups, more detailed information on supplement use, an expanded food frequency questionnaire, a different number of recall days and incorporation of diet and health knowledge questions. PMID- 12566509 TI - Collection of food and dietary supplement intake data: What We Eat in America NHANES. AB - This paper describes the collection process for the integrated dietary component of the National Health and Nutrition Examination Survey(NHANES) 2002 (entitled What We Eat in America-NHANES), referred to here as the integrated survey. The dietary components of previous NHANES cycles and the Continuing Survey of Food Intake in Individuals (CSFII) are also described. The collection process for foods in the integrated survey consists of an in-person 24-h recall using a computerized 5-step method and a second nonconsecutive 24-h recall via telephone. A food frequency questionnaire is being pilot-tested to provide information on the propensity to consume certain foods. Dietary supplement intakes over the past 30 d are assessed for all persons during the household interview. Other diet related data are also obtained. Strengths of the integrated survey include information on food and supplement intakes in a representative sample of the civilian noninstitutionalized population of the United States that can be linked to anthropometric, biochemical, clinical and disease history information in NHANES. After reviewing the current state of the art on dietary and dietary supplement data collection, discussion groups consisting of members of key stakeholder community concluded that, although the most advanced methods for dietary data collection available are being used, the differences between how information on food and dietary supplement intakes is collected make it challenging to combine data describing nutrients from both sources to obtain estimates of total nutrient intakes. The discussion groups concluded that more research is needed on these issues and provided key recommendations for future efforts in this important area of public health surveillance. PMID- 12566510 TI - Estimation of usual intake distributions of nutrients and foods. AB - The issue of estimating usual intake distributions using daily intake data as collected by nationwide food consumption surveys is discussed. Of interest are not only the usual nutrient intake distributions based on food intake alone, but also the total nutrient intake distributions that must be based on information on food and supplement consumption. The problems of estimating usual food intake distributions and distinguishing between frequently consumed and infrequently consumed food items are considered. Data needs as well as statistical methodologies available to carry out each of the tasks outlined above are discussed, with particular reference to the integrated National Health and Nutrition Examination Survey that is now in the field. The replicated 24-h recalls should be augmented with a propensity questionnaire to improve on the estimation of intake distributions for infrequently consumed nutrients, supplements and food items. PMID- 12566511 TI - Estimation of usual intakes: What We Eat in America-NHANES. AB - Usual intakes of nutrients are reliable indicators for making associations between diet and health or disease risks. Estimates of consumption of specific foods and food groups are also important for evaluating the progress in meeting key objectives in such national public health initiatives as Healthy People 2010. Reliable and valid estimates of intakes of particular foods, food ingredients, dietary supplements and other bioactive substances are also needed for dietary assessment and regulatory purposes. The ability to generate useful estimates of these constituents often requires much larger sample sizes than are needed for estimating nutrient intakes. Statistical methods recommended by the National Academy of Sciences are described that provide estimates of distributions of usual nutrient intakes and permit dietary assessment and planning at the population level. Statistical and modeling approaches for estimating intakes of foods, dietary supplements and other bioactive substances are also summarized. Based on the deliberations of discussion groups consisting of members of key stakeholder groups involved in the planning, implementation and utilization of national survey data, a high priority was placed on the need for more research to determine the best approaches for applying these methods to dietary data in the integrated What We Eat in America-National Health and Nutrition Examination Survey (NHANES). PMID- 12566512 TI - Food and dietary supplement databases for What We Eat in America-NHANES. AB - Relative strengths and potential approaches for improvement of food and dietary supplement databases used for tabulating intakes from the dietary component of the What We Eat in America-National Health and Nutrition Examination Survey (NHANES) are discussed. The U.S. Department of Agriculture's Nutrient Data Laboratory develops and maintains the Nutrient Databank System (NDBS) and many nutrient-specific and population-specific databases. NDBS contains data for approximately 8,000 foods and approximately 115 components; tables for compounds of special interest are also available. Nutrient databases need constant revision because of a constantly changing food supply. The completeness of analytical data varies from nutrient to nutrient. The National Center for Health Statistics developed and maintains a database of dietary supplements based on label information. To date, no verification of ingredients has been undertaken. The development of a dietary supplement database containing analytical values would require extensive resources but would be valuable. Databases for vitamin and mineral supplements are compatible with food databases. Databases for botanicals and other supplements include nonnutrient constituents that may not be documented in food composition databases. Gaps in food and dietary supplement composition data exist because of limited resources, changing availability of foods and products and the advent of new compounds of health interest. More data are needed on nutrients and other bioactive constituents in foods and dietary supplements. Analytical methods do not exist for all ingredients or active constituents in foods and dietary supplements. Research needs for further development of meaningful food and dietary supplement databases are similar. PMID- 12566514 TI - Science in motion: common molecular pathological themes emerge in the hereditary spastic paraplegias. AB - The hereditary spastic paraplegias are a group of neurodegenerative conditions that all share the principal clinical feature of progressive lower limb spastic paralysis, caused by either failure of development or progressive degeneration of the corticospinal tract. The conditions are characterised by extreme genetic heterogeneity, with at least 20 genes involved. Until recently, no functional overlap was apparent in the associated molecular pathological mechanisms. However, with recent progress in hereditary spastic paraplegia gene identification, common pathological themes are now emerging. PMID- 12566516 TI - Angelman syndrome: a review of the clinical and genetic aspects. AB - Angelman syndrome (AS) is a neurodevelopmental disorder characterised by severe learning difficulties, ataxia, a seizure disorder with a characteristic EEG, subtle dysmorphic facial features, and a happy, sociable disposition. Most children present with delay in developmental milestones and slowing of head growth during the first year of life. In the majority of cases speech does not develop. Patients with AS have a characteristic behavioural phenotype with jerky movements, frequent and sometimes inappropriate laughter, a love of water, and sleep disorder. The facial features are subtle and include a wide, smiling mouth, prominent chin, and deep set eyes. It is caused by a variety of genetic abnormalities involving the chromosome 15q11-13 region, which is subject to genomic imprinting. These include maternal deletion, paternal uniparental disomy, imprinting defects, and point mutations or small deletions within the UBE3A gene, which lies within this region. UBE3A shows tissue specific imprinting, being expressed exclusively from the maternal allele in brain. The genetic mechanisms identified so far in AS are found in 85-90% of those with the clinical phenotype and all interfere with UBE3A expression. PMID- 12566517 TI - Effect of TNF and LTA polymorphisms on biological markers of response to oxidative stimuli in coal miners: a model of gene-environment interaction. Tumour necrosis factor and lymphotoxin alpha. AB - INTRODUCTION: Interaction between genetic background and oxidative environmental stimuli in the pathogenesis of human lung disease has been largely unexplored. METHODS: A prospective epidemiological study was undertaken in 253 coal miners. Intermediate quantitative phenotypes of response to oxidant exposure, including erythrocyte glutathione peroxidase (GSH-Px) and catalase activities, were studied. Oxidant exposures studied were smoking habits and cumulative dust exposure assessed by job history and ambient measures. Disease phenotypes included subclinical computed tomography score at the first survey and x ray profusion grades twice, five years apart, to assess established coal workers' pneumoconiosis (CWP). Miners were genotyped for common functional polymorphisms in the gene for tumour necrosis factor alpha (TNF) and lymphotoxin alpha (LTA), two proinflammatory cytokines that have been implicated in the pathogenesis of chronic lung diseases. RESULTS: Regarding gene-environment interaction on intermediate phenotypes, results showed interaction of a promoter polymorphism at the -308 position in TNF with occupational exposure on erythrocyte GSH-Px activity with a significant association in those with high exposure (p=0.003), whereas no association was observed among those with low exposure (interaction p=0.06). Regarding gene intermediate phenotype interaction on clinical outcome, results showed an association of CWP prevalence with an NcoI polymorphism in LTA in those with low catalase activity (p=0.05), whereas no association was observed in those with high activity (interaction p=0.03). No other significant association was observed. CONCLUSION: The results suggest that interactions of genetic background with environmental exposure and intermediate response phenotypes are important components in the pathogenesis of CWP. PMID- 12566518 TI - Association of the CD14 gene -159C polymorphism with progression of IgA nephropathy. AB - The risk factors associated with the progression of IgA nephropathy (IgAN), the most common form of glomerulonephritis, are unclear. It has been suggested that CD14 signalling in response to various microbes affects the natural history of chronic inflammatory conditions. It has been hypothesised that variants in the promoter region of the CD14 gene might alter the expression of CD14, and this in turn could influence the progressive nature of IgAN. PCR-RFLP was used to determine the polymorphism at the -159 site (T to C). The distribution of the CD14/-159 polymorphism was no different in patients with IgAN (n=216) compared to 171 healthy controls. After follow up for 86 months, it was found that an excess of the C genotype occurred in patients with progressive disease (p=0.03) and the risk of disease progression increased as the number of C alleles increased (p for trend = 0.002). The hazard ratio for progression in the patients with the CC genotype was 3.2 (p=0.025) compared with the patients possessing the TT genotype. After LPS stimulation, sCD14 was released more abundantly from the PBMCs of the TT subjects than from that of the CC subjects (p=0.006), even though mCD14 expression level was no different. In addition, the TT subjects released less IL 6 than the CC subjects after stimulation (p=0.0003). These results suggest that the CD14/-159 polymorphism is an important marker for the progression of IgAN and may modulate the level of the inflammatory responses. PMID- 12566519 TI - Molecular study of frequency of mosaicism in neurofibromatosis 2 patients with bilateral vestibular schwannomas. AB - Neurofibromatosis 2 (NF2) is a severe autosomal dominant disorder that predisposes to multiple tumours of the nervous system. About half of all patients are founders with clinically unaffected parents. The purpose of the present study was to examine the extent to which mosaicism is present in NF2 founders. A total of 233 NF2 founders with bilateral vestibular schwannomas (BVS) were screened by exon scanning. NF2 mutations were detected in the blood samples of 122 patients (52%). In 10 of the 122 cases, the ratio of mutant to normal alleles was obviously less than 1, suggesting mosaicism. Tumour specimens were available from 35 of the 111 subjects in whom no mutation could be detected in blood specimens. Mutational analysis by exon scanning detected typical NF2 mutations in 21 of the 35 tumours. In nine subjects, the alterations found in tumours could be confirmed to be the constitutional mutation based on finding of identical mutations in pathologically and/or anatomically distinct second tumours. In six other subjects with only a single tumour available, allelic loss of the NF2 gene was found in addition to the mutation in each tumour, suggesting that either the mutation or the deletion of the NF2 gene is probably the constitutional genetic alteration. Our results suggest that failure to find constitutional mutations in blood specimen from these 15 patients was not because of the limitation of the applied screening technique, but the lack of the mutations in their leucocytes, best explained by mosaicism. Extrapolating the rate (15/35 = 43%) of mosaicism in these 35 cases to the 111 NF2 founders with no constitutional NF2 mutations found in their blood, we inferred 48 mosaic subjects (111 x 0.429). Adding the 10 mosaic cases detected directly in blood specimens, we estimate the rate of mosaicism to be 24.8% (58/233) in our cohort of 233 NF2 founders with bilateral vestibular schwannomas. PMID- 12566520 TI - A phenocopy of CAII deficiency: a novel genetic explanation for inherited infantile osteopetrosis with distal renal tubular acidosis. AB - The rare bone thickening disease osteopetrosis occurs in various forms, one of which is accompanied by renal tubular acidosis (RTA), and is known as Guibaud Vainsel syndrome or marble brain disease. Clinical manifestations of this autosomal recessive syndrome comprise increased bone density, growth failure, intracerebral calcification, facial dysmorphism, mental retardation, and conductive hearing impairment. The most common cause is carbonic anhydrase II (CAII) deficiency. Several different loss of function mutations in CA2, the gene encoding CAII, have been described. To date, there have been no exceptions to the finding of CAII deficiency in patients with coexistent osteopetrosis and RTA. Most often, the RTA is of mixed proximal and distal type, but kindreds are reported in which either distal or proximal RTA predominates. We report the molecular genetic investigation of two consanguineous kindreds where osteopetrosis and distal RTA (dRTA) were both manifest. One kindred harbours a novel homozygous frameshift alteration in CA2. In the other, CAII levels were normal despite a similar clinical picture, and we excluded defects in CA2. In this kindred, two separate recessive disorders are penetrant, each affecting a different, tissue specific subunit of the vacuolar proton pump (H(+)-ATPase), providing a highly unusual, novel genetic explanation for the coexistence of osteopetrosis and dRTA. The osteopetrosis is the result of a homozygous deletion in TCIRG1, which encodes an osteoclast specific isoform of subunit a of the H(+) ATPase, while the dRTA is associated with a homozygous mutation in ATP6V1B1, encoding the kidney specific B1 subunit of H(+)-ATPase. This kindred is exceptional firstly because the coinheritance of two rare recessive disorders has created a phenocopy of CAII deficiency, and secondly because these disorders affect two different subunits of the H(+)-ATPase that have opposite effects on bone density, but which have only recently been determined to possess tissue specific isoforms. PMID- 12566521 TI - Independent NF1 mutations in two large families with spinal neurofibromatosis. PMID- 12566522 TI - A complex deletion-inversion-deletion event results in a chimeric IL1RAPL1 dystrophin transcript and a contiguous gene deletion syndrome. PMID- 12566523 TI - Mild brachydactyly type A1 maps to chromosome 2q35-q36 and is caused by a novel IHH mutation in a three generation family. PMID- 12566524 TI - Using case study comparisons to explore genotype-phenotype correlations in Williams-Beuren syndrome. PMID- 12566526 TI - Relationship between microsatellite polymorphism in the haem oxygenase-1 gene promoter and longevity of the normal Japanese population. PMID- 12566525 TI - The use of genotype-phenotype correlations in mutation analysis for the long QT syndrome. PMID- 12566528 TI - Association of germline mutations and polymorphisms of the RET proto-oncogene with idiopathic congenital central hypoventilation syndrome in 33 patients. PMID- 12566529 TI - SHOX mutations detected by FISH and direct sequencing in patients with short stature. PMID- 12566530 TI - Genotype-phenotype correlations in 35 Brazilian families with sarcoglycanopathies including the description of three novel mutations. PMID- 12566531 TI - Developmental delay and the methyl binding genes. PMID- 12566532 TI - Mutational analysis of N-ras, p53, CDKN2A (p16(INK4a)), p14(ARF), CDK4, and MC1R genes in human dysplastic melanocytic naevi. PMID- 12566533 TI - A novel pericentric inversion of chromosome 3 cosegregates with a developmental behavioural phenotype. PMID- 12566534 TI - Appraisal of genetic and epigenetic congruity of a monozygotic twin pair discordant for schizophrenia. PMID- 12566535 TI - Large deletion causing the TSC2-PKD1 contiguous gene syndrome without infantile polycystic disease. PMID- 12566536 TI - The 56th Annual Meeting and Symposium of the Society of General Physiologists: trafficking of transporters. PMID- 12566537 TI - Gating mechanism of BK (Slo1) channels: so near, yet so far. PMID- 12566538 TI - Titin determines the Frank-Starling relation in early diastole. AB - Titin, a giant protein spanning half the sarcomere, is responsible for passive and restoring forces in cardiac myofilaments during sarcomere elongation and compression, respectively. In addition, titin has been implicated in the length dependent activation that occurs in the stretched sarcomere, during the transition from diastole to systole. The purpose of this study was to investigate the role of titin in the length-dependent deactivation that occurs during early diastole, when the myocyte is shortened below slack length. We developed a novel in vitro assay to assess myocyte restoring force (RF) by measuring the velocity of recoil in Triton-permeabilized, unloaded rat cardiomyocytes after rigor induced sarcomere length (SL) contractions. We compared rigor-induced SL shortening to that following calcium-induced (pCa) contractions. The RF-SL relationship was linearly correlated, and the SL-pCa curve displayed a characteristic sigmoidal curve. The role of titin was defined by treating myocytes with a low concentration of trypsin, which we show selectively degrades titin using mass spectroscopic analysis. Trypsin treatment reduced myocyte RF as shown by a decrease in the slope of the RF-SL relationship, and this was accompanied by a downward and leftward shift of the SL-pCa curve, indicative of sensitization of the myofilaments to calcium. In addition, trypsin digestion did not alter the relationship between SL and interfilament spacing (assessed by cell width) after calcium activation. These data suggest that as the sarcomere shortens below slack length, titin-based restoring forces act to desensitize the myofilaments. Furthermore, in contrast to length-dependent activation at long SLs, length-dependent deactivation does not depend on interfilament spacing. This study demonstrates for the first time the importance of titin-based restoring force in length-dependent deactivation during the early phase of diastole. PMID- 12566539 TI - Influence of pore residues on permeation properties in the Kv2.1 potassium channel. Evidence for a selective functional interaction of K+ with the outer vestibule. AB - The Kv2.1 potassium channel contains a lysine in the outer vestibule (position 356) that markedly reduces open channel sensitivity to changes in external [K(+)]. To investigate the mechanism underlying this effect, we examined the influence of this outer vestibule lysine on three measures of K(+) and Na(+) permeation. Permeability ratio measurements, measurements of the lowest [K(+)] required for interaction with the selectivity filter, and measurements of macroscopic K(+) and Na(+) conductance, were all consistent with the same conclusion: that the outer vestibule lysine in Kv2.1 interferes with the ability of K(+) to enter or exit the extracellular side of the selectivity filter. In contrast to its influence on K(+) permeation properties, Lys 356 appeared to be without effect on Na(+) permeation. This suggests that Lys 356 limited K(+) flux by interfering with a selective K(+) binding site. Combined with permeation studies, results from additional mutagenesis near the external entrance to the selectivity filter indicated that this site was located external to, and independent from, the selectivity filter. Protonation of a naturally occurring histidine in the same outer vestibule location in the Kv1.5 potassium channel produced similar effects on K(+) permeation properties. Together, these results indicate that a selective, functional K(+) binding site (e.g., local energy minimum) exists in the outer vestibule of voltage-gated K(+) channels. We suggest that this site is the location of K(+) hydration/dehydration postulated to exist based on the structural studies of KcsA. Finally, neutralization of position 356 enhanced outward K(+) current magnitude, but did not influence the ability of internal K(+) to enter the pore. These data indicate that in Kv2.1, exit of K(+) from the selectivity filter, rather than entry of internal K(+) into the channel, limits outward current magnitude. We discuss the implications of these findings in relation to the structural basis of channel conductance in different K(+) channels. PMID- 12566540 TI - Inactivation of BK channels by the NH2 terminus of the beta2 auxiliary subunit: an essential role of a terminal peptide segment of three hydrophobic residues. AB - An auxiliary beta2 subunit, when coexpressed with Slo alpha subunits, produces inactivation of the resulting large-conductance, Ca(2+) and voltage-dependent K(+) (BK-type) channels. Inactivation is mediated by the cytosolic NH(2) terminus of the beta2 subunit. To understand the structural requirements for inactivation, we have done a mutational analysis of the role of the NH(2) terminus in the inactivation process. The beta2 NH(2) terminus contains 46 residues thought to be cytosolic to the first transmembrane segment (TM1). Here, we address two issues. First, we define the key segment of residues that mediates inactivation. Second, we examine the role of the linker between the inactivation segment and TM1. The results show that the critical determinant for inactivation is an initial segment of three amino acids (residues 2-4: FIW) after the initiation methionine. Deletions that scan positions from residue 5 through residue 36 alter inactivation, but do not abolish it. In contrast, deletion of FIW or combinations of point mutations within the FIW triplet abolish inactivation. Mutational analysis of the three initial residues argues that inactivation does not result from a well-defined structure formed by this epitope. Inactivation may be better explained by linear entry of the NH(2)-terminal peptide segment into the permeation pathway with residue hydrophobicity and size influencing the onset and recovery from inactivation. Examination of the ability of artificial, polymeric linkers to support inactivation suggests that a variety of amino acid sequences can serve as adequate linkers as long as they contain a minimum of 12 residues between the first transmembrane segment and the FIW triplet. Thus, neither a specific distribution of charge on the linker nor a specific structure in the linker is required to support the inactivation process. PMID- 12566541 TI - Involvement of helices at the dimer interface in ClC-1 common gating. AB - ClC-1 is a dimeric, double-pored chloride channel that is present in skeletal muscle. Mutations of this channel can result in the condition myotonia, a muscle disorder involving increased muscle stiffness. It has been shown that the dominant form of myotonia often results from mutations that affect the so-called slow, or common, gating process of the ClC-1 channel. Mutations causing dominant myotonia are seen to cluster at the interface of the ClC-1 channel monomers. This study has investigated the role of the H, I, P, and Q helices, which lie on this interface, as well as the G helix, which is situated immediately behind the H and I helices, on ClC-1 gating. 11 mutant ClC-1 channels (T268M, C277S, C278S, S289A, T310M, S312A, V321S, T539A, S541A, M559T, and S572V) were produced using site directed mutagenesis, and gating properties of these channels were investigated using electrophysiological techniques. Six of the seven mutations in G, H, and I, and two of the four mutations in P and Q, caused shifts of the ClC-1 open probability. In the majority of cases this was due to alterations in the common gating process, with only three of the mutants displaying any change in fast gating. Many of the mutant channels also showed alterations in the kinetics of the common gating process, particularly at positive potentials. The changes observed in common gating were caused by changes in the opening rate (e.g. T310M), the closing rate (e.g. C277S), or both rates. These results indicate that mutations in the helices forming the dimer interface are able to alter the ClC-1 common gating process by changing the energy of the open and/or closed channel states, and hence altering transition rates between these states. PMID- 12566543 TI - Medicine in the Falkland Islands. PMID- 12566542 TI - Molecular action of lidocaine on the voltage sensors of sodium channels. AB - Block of sodium ionic current by lidocaine is associated with alteration of the gating charge-voltage (Q-V) relationship characterized by a 38% reduction in maximal gating charge (Q(max)) and by the appearance of additional gating charge at negative test potentials. We investigated the molecular basis of the lidocaine induced reduction in cardiac Na channel-gating charge by sequentially neutralizing basic residues in each of the voltage sensors (S4 segments) in the four domains of the human heart Na channel (hH1a). By determining the relative reduction in the Q(max) of each mutant channel modified by lidocaine we identified those S4 segments that contributed to a reduction in gating charge. No interaction of lidocaine was found with the voltage sensors in domains I or II. The largest inhibition of charge movement was found for the S4 of domain III consistent with lidocaine completely inhibiting its movement. Protection experiments with intracellular MTSET (a charged sulfhydryl reagent) in a Na channel with the fourth outermost arginine in the S4 of domain III mutated to a cysteine demonstrated that lidocaine stabilized the S4 in domain III in a depolarized configuration. Lidocaine also partially inhibited movement of the S4 in domain IV, but lidocaine's most dramatic effect was to alter the voltage dependent charge movement of the S4 in domain IV such that it accounted for the appearance of additional gating charge at potentials near -100 mV. These findings suggest that lidocaine's actions on Na channel gating charge result from allosteric coupling of the binding site(s) of lidocaine to the voltage sensors formed by the S4 segments in domains III and IV. PMID- 12566544 TI - Drotrecogin alfa (activated): a novel therapeutic strategy for severe sepsis. AB - Recent studies have highlighted the close link between activation of the coagulation system and the inflammatory response in the pathophysiology of severe sepsis. The protein C anticoagulant pathway plays an integral part in modulating the coagulation and inflammatory responses to infection. In patients with sepsis, endogenous protein C levels are decreased, shifting the balance toward greater systemic inflammation, coagulation, and cell death. On the basis of a single large randomised phase 3 trial, drotrecogin alfa (activated), a recombinant form of human activated protein C, was recently approved for the treatment of adult patients with severe sepsis and a high risk of death. Since its approval, several questions have been raised regarding the appropriate use of this agent. Given the increased risk of serious bleeding and the high cost of treatment, drotrecogin alfa (activated) should be reserved at this time for the most acutely ill patients with severe sepsis who meet the criteria that were used in the phase 3 trial. PMID- 12566545 TI - Acute disseminated encephalomyelitis. AB - Acute disseminated encephalomyelitis (ADEM) is an acute demyelinating disorder of the central nervous system, and is characterised by multifocal white matter involvement. Diffuse neurological signs along with multifocal lesions in brain and spinal cord characterise the disease. Possibly, a T cell mediated autoimmune response to myelin basic protein, triggered by an infection or vaccination, underlies its pathogenesis. ADEM is a monophasic illness with favourable long term prognosis. The differentiation of ADEM from a first attack of multiple sclerosis has prognostic and therapeutic implications; this distinction is often difficult. Most patients with ADEM improve with methylprednisolone. If that fails immunoglobulins, plasmapheresis, or cytotoxic drugs can be given. Recent literature suggests that a significant proportion of patients with ADEM will later develop multiple sclerosis; however, follow up experience from developing countries does not support this view. PMID- 12566546 TI - Hereditary haemorrhagic telangiectasia (Osler-Weber-Rendu syndrome): a view from the 21st century. AB - Hereditary haemorrhagic telangiectasia (HHT) affects one in 5-8000, and no longer can be viewed as solely causing anaemia (due to nasal and gastrointestinal bleeding) and characteristic mucocutaneous telangiectasia. Arteriovenous malformations commonly occur, and in the pulmonary and cerebral circulations demand knowledge of risks and benefits of asymptomatic screening and treatment. HHT is inherited as an autosomal dominant trait and there is no age cut off when apparently unaffected offspring of an individual with HHT can be told they are unaffected. This review focuses on the evolving evidence base for HHT management, issues regarding pregnancy and prothrombotic treatments, and discusses the molecular and cellular changes that underlie this disease. PMID- 12566547 TI - Evaluation and treatment of dyspepsia. AB - Dyspepsia is a common symptom. Dyspeptic symptoms may be caused by a variety of conditions such as peptic ulcer disease, gastro-oesophageal reflux, and malignancy. Most often, however, no cause is identified and dyspepsia is deemed to be functional. While symptom severity does influence frequency of consultation, dyspeptic consulters also differ from non-consulters with respect to symptom perception and anxiety. This highlights the importance of understanding the patient's agenda early in the course of evaluation. Patients over the age of 55 years or with alarm symptoms should be referred for prompt endoscopy. In the absence of other clinically apparent aetiologies, uninvestigated dyspeptics can be either tested and treated for Helicobacter pylori or empirically treated with proton pump inhibitors. Uninvestigated dyspeptics failing empiric therapy should be referred for evaluation that includes endoscopy. Further therapy with prokinetics, tricyclic antidepressants, fundal relaxants, antidepressants, or psychotherapy is guided by predominant symptoms and assessment of possible psychiatric factors. PMID- 12566548 TI - Perichondritis: a complication of piercing auricular cartilage. PMID- 12566549 TI - Evolution of diabetes medical nutrition therapy. AB - Research supports the importance of medical nutrition therapy in achieving diabetes treatment goals. For persons requiring insulin therapy, the first priority is to integrate an insulin regimen into the patient's lifestyle. For type 2 diabetes, the priority is to focus on lifestyle strategies (that is, nutrition and exercise) that will improve metabolic outcomes at diagnosis and as the disease progresses. Patients with diabetes need nutrition recommendations that are supported by scientific evidence and that can be easily understood and translated into everyday life. To achieve positive outcomes, a coordinated team effort that provides continued education and support is essential. PMID- 12566550 TI - HIV viral suppression in the era of antiretroviral therapy. AB - Altogether 42 million people worldwide have been infected with HIV, and 12 million have died over the last 20 years. Effective antiretroviral therapy has lead to sustained HIV viral suppression and immunological recovery in patients who have been infected with the virus. The incidence of AIDS has declined in the Western world with the introduction of effective antiretroviral therapy. Questions on When to start treatment?, What to start with?, How to monitor patients?, remain heavily debated. Adherence to antiretroviral treatment remains the cornerstone of effective treatment, and failure to adhere is the strongest predictor of virological failure. Long term therapy can lead to metabolic complications. Resource poor countries are dealing with difficult issues such as mother to child prevention of HIV transmission. Other treatment options are now available, with the recent introduction of fusion inhibitors, second generation non-nucleoside reverse transcriptase inhibitors, and nucleotide reverse transcriptase inhibitors to clinical practice. PMID- 12566552 TI - Total arterial revascularisation as a primary strategy for coronary artery bypass grafting. AB - BACKGROUND: Bilateral internal thoracic arteries confer improved survival benefit after coronary artery bypass grafting (CABG). Despite increasing evidence, the use of arterial conduits has not been accepted as a primary practice in most of the centres in the UK for various reasons. A series of patients has been analysed to assess the feasibility of total arterial revascularisation as a primary strategy in patients requiring first time CABG. METHODS: Altogether 245 patients undergoing first time CABG by one surgeon, from June 1999 to October 2000, were studied. Group 1 consisted of 165 patients undergoing total arterial revascularisation (using bilateral internal thoracic and radial arteries) and group 2 consisted of 80 patients undergoing conventional CABG (using one internal thoracic artery and supplemental veins). Thirty day mortality and early morbidity with particular reference to resternotomy for bleeding, cerebrovascular accidents, renal failure, and sternal dehiscence were the main outcome measures. RESULTS: Patients in group 1 were younger (mean (SD) 60 (10) v 65 (9) years; p<0.001), had lower Parsonnet scores (mean (SD) 5 (5) v 11 (7); p<0.001), and better left ventricular function. Both groups received a similar number of grafts. The percentage of patients undergoing total arterial revascularisation rose from 44% in the first three months to over 75% in the three latter three month periods. Overall 30 day mortality was 1.3%, one patient (0.6%) in group 1 and two patients (2.5%) in group 2. There was a similar incidence of postoperative complications and length of median postoperative stay in both groups. CONCLUSION: Total arterial revascularisation can be adopted as a primary strategy in most patients undergoing CABG with no increase in mortality or morbidity. PMID- 12566553 TI - Pupillary evaluation for differential diagnosis of coma. AB - OBJECTIVES: To determine the usefulness of bedside evaluation of pupils in determining the aetiology of coma by adopting a probabilistic approach. PATIENTS AND METHODS: One hundred and fifteen consecutive patients presenting with coma were enrolled in this prospective cohort during the 12 month study period in the emergency room of a community teaching hospital. Patients underwent structured clinical examinations and laboratory and imaging tests. Assignment of aetiology of coma was based on strict adherence to predetermined criteria and achieved by consensus of the two physician investigators. One year follow up was obtained in all patients. RESULTS: Aetiology of coma was determined in 98% of the patients. It was metabolic in 69 patients (60%) and structural in 46 patients (40%). Metabolic causes included drug overdose, acute alcohol intoxication, hypoglycaemia, sepsis, and pneumonia. Structural causes included intracerebral haemorrhage, subarachnoid haemorrhage, cerebral infarction, subdural haematoma, and epidural haematoma. Multivariate logistic regression analysis showed light reflex loss (likelihood ratio for positive test result 3.59) and anisocoria (likelihood ratio for positive test result 9.0) as independent predictors of structural origin. CONCLUSIONS: In this prospective study of patients presenting to the emergency room of a community based teaching hospital with coma, in about 60% the coma is of metabolic origins and in about 40% of structural origins. Light reflex loss and anisocoria suggest a structural aetiology. PMID- 12566554 TI - Are medical reports on fitness to drive trustworthy? AB - PURPOSE: Previous studies have shown that physicians often have poor knowledge of the medical restriction on fitness to drive, or submit poor quality medical reports. To determine the reliability of physicians' reports on fitness to drive, the medical data provided on the standard medical fitness form was compared with the additional data collected on repeated assessment. METHODS: A random sample of 100 applicants for a driver's licence aged more than 49 years who submitted the standard medical form were asked to provide, from their regular family doctor, confirmation of their health status and/or additional medical data in order to make a re-evaluation. RESULTS: The rate of rejection for a licence for medical reasons was 3% on the basis of the standard evaluation and 17% on the basis of the re-evaluation (p<0.001). CONCLUSION: This study shows that the random evaluation of physician assessments of applicants for a driver's licence increases the detection rate of medical problems that can affect fitness to drive. The alarming difference in the rate of rejection between the two assessments may reflect a lackadaisical attitude of medical professionals toward the licence assessment procedure and/or physician unawareness of the medical restrictions on fitness to drive. Results of this study suggest that this subject must be included in medical education programmes. PMID- 12566555 TI - Unusual complications of thyroid carcinoma. AB - Thyroid carcinomas are the most frequent endocrine malignancies. Complications may arise from an established malignancy and these may lead to the initial clinical presentation or cause subsequent problems. In this case report two elderly patients with differentiated and undifferentiated thyroid carcinomas who suffered unusual medical complications are described. The incidence and treatment of thyroid carcinoma is discussed and the importance of fine needle aspiration of thyroid nodules and the recognition of iodine containing radiographic contrast media in the causation of iodine induced thyrotoxicosis is highlighted. PMID- 12566556 TI - Late small bowel obstruction after blunt abdominal trauma. AB - Blunt abdominal trauma with intraperitoneal injury usually presents acutely. On rare occasions, such patients can present later on with features of small bowel obstruction due to stricture formation. It is thought that such a delayed stricture is due to subclinical bowel perforation, localised gut ischaemia, or injury to the mesenteric vasculature. This case demonstrates the mesenteric vascular injury theory to be the cause of the bowel stricture. PMID- 12566557 TI - Phantom lymphadenopathy. An association with chronic fatigue syndrome. AB - Ten patients with self diagnosed enlarged lymph glands were referred to a general medicine outpatient clinic and careful examination did not confirm lymphadenopathy. All patients also complained of severe chronic fatigue associated with aches and miscellaneous somatic symptoms, and fulfilled criteria for diagnosis of chronic fatigue syndrome (CFS). Phantom lymphadenopathy may be a symptom in some people with CFS, and possible reasons for this are discussed. PMID- 12566558 TI - Hyponatraemia in a woman with a pelvic mass. PMID- 12566559 TI - Multiple pulmonary nodules: diagnosis in an young afebrile patient. PMID- 12566562 TI - Chemosensory regulation of developmental gene expression in Myxococcus xanthus. AB - The delta-proteobacterium Myxococcus xanthus coordinates its motility during aggregation and fruiting body formation. While searching for chemotaxis genes in M. xanthus, we identified a third chemotaxis-like gene cluster, the che3 cluster, encoding homologs to two methyl-accepting chemotaxis proteins (MCPs), a CheW, a hybrid CheA, a CheB, a CheR, but no CheY. Mutations in mcp3A, mcp3B, and cheA3 did not show obvious defects in motility or chemotaxis but did affect the timing of entry into development. Mutations in these genes caused early aggregation of starving cells, even at low cell densities. Furthermore, these mutants showed pronounced overexpression of the developmentally regulated Tn5lac fusions Omega4403, Omega4411, and Omega4521 as well as overexpression of mbhA and tps, markers for peripheral rods and aggregating cells, respectively. Divergently transcribed from the che3 promoter region is another gene, crdA (chemosensory regulator of development), predicted to encode a transcriptional activator of sigma(54)-dependent promoters. To test the hypothesis that CrdA functions as the cognate response regulator for the histidine kinase CheA3, CrdA and CheA3 were assayed and found to interact strongly in the yeast two-hybrid system. Mutant analysis showed that crdA cells were delayed in development (12-24 h) and delayed in MbhA production relative to the wild type. An mcp3BcrdA double mutant displayed the crdA phenotype, indicating that crdA is epistatic to mcp3B. We conclude that the Che3 chemotaxis-like system functions to control developmental gene expression by regulating a sigma(54) transcriptional activator, CrdA. PMID- 12566563 TI - Social exploitation of vitellogenin. AB - Vitellogenin is a female-specific glucolipoprotein yolk precursor produced by all oviparous animals. Vitellogenin expression is under hormonal control, and the protein is generally synthesized directly before yolk deposition. In the honeybee (Apis mellifera), vitellogenin is not only synthesized by the reproductive queen, but also by the functionally sterile workers. In summer, the worker population consists of a hive bee group performing a multitude of tasks including nursing inside the nest, and a forager group specialized in collecting nectar, pollen, water, and propolis. Vitellogenin is synthesized in large quantities by hive bees. When hive bees develop into foragers, their juvenile hormone titers increase, and this causes cessation of their vitellogenin production. This inverse relationship between vitellogenin synthesis and juvenile hormone is opposite to the norm in insects, and the underlying proximate processes and life history reasons are still not understood. Here we document an alternative use of vitellogenin by showing that it is a source for the proteinaceous royal jelly that is produced by the hive bees. Hive bees use the jelly to feed larvae, queen, workers, and drones. This finding suggests that the evolution of a brood-rearing worker class and a specialized forager class in an advanced eusocial insect society has been directed by an alternative utilization of yolk protein. PMID- 12566564 TI - Designed to be stable: crystal structure of a consensus ankyrin repeat protein. AB - Ankyrin repeat (AR) proteins mediate innumerable protein-protein interactions in virtually all phyla. This finding suggested the use of AR proteins as designed binding molecules. Based on sequence and structural analyses, we designed a consensus AR with fixed framework and randomized interacting residues. We generated several combinatorial libraries of AR proteins consisting of defined numbers of this repeat. Randomly chosen library members are expressed in soluble form in the cytoplasm of Escherichia coli constituting up to 30% of total cellular protein and show high thermodynamic stability. We determined the crystal structure of one of those library members to 2.0-A resolution, providing insight into the consensus AR fold. Besides the highly complementary hydrophobic repeat repeat interfaces and the absence of structural irregularities in the consensus AR protein, the regular and extended hydrogen bond networks in the beta-turn and loop regions are noteworthy. Furthermore, all residues found in the turn region of the Ramachandran plot are glycines. Many of these features also occur in natural AR proteins, but not in this rigorous and standardized fashion. We conclude that the AR domain fold is an intrinsically very stable and well expressed scaffold, able to display randomized interacting residues. This scaffold represents an excellent basis for the design of novel binding molecules. PMID- 12566565 TI - Short-term administration of omega 3 fatty acids from fish oil results in increased transthyretin transcription in old rat hippocampus. AB - Reduced brain levels of long chain polyunsaturated fatty acids [arachidonic acid and docosahexanoic acid (DHA)] are observed in elderly subjects and patients with Alzheimer's disease. To determine the effects of n-3 fatty acids on aged rat brain, 2-year-old rats were fed fish oil (27% DHA content) for 1 month, and gene expression analysis and fatty acid and molecular species composition of the major phospholipid species were assessed. No significant alteration could be observed in the fatty acid composition of ethanolamine phosphoglycerides and phosphatidylserines with the exception of DHA, which was slightly higher in brains of rats receiving fish oil. However, a drastic reduction in arachidonic acid in phosphatidylinositoles was observed. The expression of 23 genes was altered in response to fish oil feeding in the hippocampus. The transcription of transthyretin (TTR) was induced by 10-fold as evidenced by microarray analysis and confirmed by real-time quantitative RT-PCR. Expression of IL-1 and NO synthase, which has been implicated in the prevention of neurological diseases, was unaltered. TTR is an amyloid beta protein scavenger, so an increase in its expression could prevent amyloid aggregate formation. We believe the beneficial effects of fish oil might be common to other agents, i.e., induce TTR expression, like nicotine and Ginkgo biloba extract. PMID- 12566566 TI - Complete genome sequence of Lactobacillus plantarum WCFS1. AB - The 3,308,274-bp sequence of the chromosome of Lactobacillus plantarum strain WCFS1, a single colony isolate of strain NCIMB8826 that was originally isolated from human saliva, has been determined, and contains 3,052 predicted protein encoding genes. Putative biological functions could be assigned to 2,120 (70%) of the predicted proteins. Consistent with the classification of L. plantarum as a facultative heterofermentative lactic acid bacterium, the genome encodes all enzymes required for the glycolysis and phosphoketolase pathways, all of which appear to belong to the class of potentially highly expressed genes in this organism, as was evident from the codon-adaptation index of individual genes. Moreover, L. plantarum encodes a large pyruvate-dissipating potential, leading to various end-products of fermentation. L. plantarum is a species that is encountered in many different environmental niches, and this flexible and adaptive behavior is reflected by the relatively large number of regulatory and transport functions, including 25 complete PTS sugar transport systems. Moreover, the chromosome encodes >200 extracellular proteins, many of which are predicted to be bound to the cell envelope. A large proportion of the genes encoding sugar transport and utilization, as well as genes encoding extracellular functions, appear to be clustered in a 600-kb region near the origin of replication. Many of these genes display deviation of nucleotide composition, consistent with a foreign origin. These findings suggest that these genes, which provide an important part of the interaction of L. plantarum with its environment, form a lifestyle adaptation region in the chromosome. PMID- 12566567 TI - Requirement of subunit expression for cAMP-mediated regulation of a heart potassium channel. AB - Beta-adrenergic receptor stimulation increases heart rate and shortens ventricular action-potential duration, the latter effect due in part to a cAMP dependent increase in the slow outward potassium current (I(Ks)). Mutations in either KCNQ1 or KCNE1, the I(Ks) subunits, are associated with variants (LQT-1 and LQT-5) of the congenital long QT syndrome. We now show that cAMP-mediated functional regulation of KCNQ1/KCNE1 channels, a consequence of cAMP-dependent protein kinase A phosphorylation of the KCNQ1 N terminus, requires coexpression of KCNQ1 with KCNE1, its auxiliary subunit. Further, at least two KCNE1 mutations linked to LQT-5 (D76N and W87R) cause functional disruption of cAMP-mediated KCNQ1/KCNE1-channel regulation despite the response of the substrate protein (KCNQ1) to protein kinase A phosphorylation. Transduction of protein phosphorylation into physiologically necessary channel function represents a previously uncharacterized role for the KCNE1 auxiliary subunit, which can be disrupted in LQT-5. PMID- 12566568 TI - Epitope and isotype specificities of antibodies to beta -amyloid peptide for protection against Alzheimer's disease-like neuropathology. AB - Transgenic PDAPP mice, which express a disease-linked isoform of the human amyloid precursor protein, exhibit CNS pathology that is similar to Alzheimer's disease. In an age-dependent fashion, the mice develop plaques containing beta amyloid peptide (Abeta) and exhibit neuronal dystrophy and synaptic loss. It has been shown in previous studies that pathology can be prevented and even reversed by immunization of the mice with the Abeta peptide. Similar protection could be achieved by passive administration of some but not all monoclonal antibodies against Abeta. In the current studies we sought to define the optimal antibody response for reducing neuropathology. Immune sera with reactivity against different Abeta epitopes and monoclonal antibodies with different isotypes were examined for efficacy both ex vivo and in vivo. The studies showed that: (i) of the purified or elicited antibodies tested, only antibodies against the N terminal regions of Abeta were able to invoke plaque clearance; (ii) plaque binding correlated with a clearance response and neuronal protection, whereas the ability of antibodies to capture soluble Abeta was not necessarily correlated with efficacy; (iii) the isotype of the antibody dramatically influenced the degree of plaque clearance and neuronal protection; (iv) high affinity of the antibody for Fc receptors on microglial cells seemed more important than high affinity for Abeta itself; and (v) complement activation was not required for plaque clearance. These results indicate that antibody Fc-mediated plaque clearance is a highly efficient and effective process for protection against neuropathology in an animal model of Alzheimer's disease. PMID- 12566569 TI - Dynamic recruitment of dynamin for final mitochondrial severance in a primitive red alga. AB - Dynamins are a eukaryote-specific family of GTPases. Some family members are involved in diverse and varied cellular activities. Here, we report that the primitive red alga Cyanidioschyzon merolae retains only one dynamin homolog, CmDnm1, belonging to the mitochondrial division subfamily. Previously, the bacterial cell division protein, FtsZ, was shown to localize at the mitochondrial division site in the alga. We showed that FtsZ and dynamin coexist as mitochondrial division-associated proteins that act during different phases of division. CmDnm1 was recruited from 10-20 cytoplasmic patches (dynamin patches) to the midpoint of the constricted mitochondrion-dividing ring (MD ring), which was observed as an electron-dense structure on the cytoplasmic side. CmDnm1 is probably not required for early constriction; it forms a ring or spiral when the outer mitochondrial membrane is finally severed, whereas the FtsZ and MD rings are formed before constriction. It is thought that the FtsZ, MD, and dynamin rings are involved in scaffolding, constriction, and final separation, respectively. In eukaryotes, mitochondrial severance is probably the most conserved role for the dynamin family. PMID- 12566570 TI - Ethanol increases GABAergic transmission at both pre- and postsynaptic sites in rat central amygdala neurons. AB - We examined the interaction of ethanol with the gamma-aminobutyric acid (GABA)ergic system in neurons of slices of the rat central amygdala nucleus (CeA), a brain region thought to be critical for the reinforcing effects of ethanol. Brief superfusion of 11-66 mM ethanol significantly increased GABA type A (GABA(A)) receptor-mediated inhibitory postsynaptic potentials (IPSPs) and currents (IPSCs) in most CeA neurons, with a low apparent EC(50) of 20 mM. Acute superfusion of 44 mM ethanol increased the amplitude of evoked GABA(A) IPSPs and IPSCs in 70% of CeA neurons. The ethanol enhancement of IPSPs and IPSCs occurred to a similar extent in the presence of the GABA type B (GABA(B)) receptor antagonist CGP 55845A, suggesting that this receptor is not involved in the ethanol effect on CeA neurons. Ethanol superfusion also decreased paired-pulse facilitation of evoked GABA(A) IPSPs and IPSCs and always increased the frequency and sometimes the amplitude of spontaneous miniature GABA(A) IPSCs as well as responses to local GABA application, indicating both presynaptic and postsynaptic sites of action for ethanol. Thus, the CeA is the first brain region to reveal, without conditional treatments such as GABA(B) antagonists, consistent, low-dose ethanol enhancement of GABAergic transmission at both pre- and postsynaptic sites. These findings add further support to the contention that the ethanol-GABA interaction in CeA plays an important role in the reinforcing effects of ethanol. PMID- 12566571 TI - Interference of hepatitis C virus RNA replication by short interfering RNAs. AB - Hepatitis C virus (HCV) infection is a major cause of chronic liver disease, which can lead to the development of liver cirrhosis and hepatocellular carcinoma. Current therapy of patients with chronic HCV infection includes treatment with IFNalpha in combination with ribavirin. Because most treated patients do not resolve the infection, alternative treatment is essential. RNA interference (RNAi) is a recently discovered antiviral mechanism present in plants and animals that induces double-stranded RNA degradation. Using a selectable subgenomic HCV replicon cell culture system, we have shown that RNAi can specifically inhibit HCV RNA replication and protein expression in Huh-7 cells that stably replicate the HCV genome, and that this antiviral effect is independent of IFN. These results suggest that RNAi may represent a new approach for the treatment of persistent HCV infection. PMID- 12566572 TI - A new twist on transposons: the maize genome harbors helitron insertion. PMID- 12566573 TI - RNA interference, transposons, and the centromere. PMID- 12566574 TI - Misexpression of the cyclin-dependent kinase inhibitor ICK1/KRP1 in single-celled Arabidopsis trichomes reduces endoreduplication and cell size and induces cell death. AB - A positive correlation between cell size and DNA content has been recognized in many plant cell types. Conversely, misexpression of a dominant-negative cyclin dependent kinase (CDK) or CDK inhibitor proteins (ICK/KRPs) in Arabidopsis and tobacco leaves has revealed that cell growth can be uncoupled from cell cycle progression and DNA content. However, cell growth also appears to be controlled in a non-cell-autonomous manner by organ size, making it difficult in a ubiquitous expression assay to judge the cell-autonomous function of putative cell growth regulators. Here, we investigated the function of the CDK inhibitor ICK1/KRP1 on cell growth and differentiation independent of any compensatory influence of an organ context using Arabidopsis trichomes as a model system. By analyzing cell size with respect to DNA content, we dissected cell growth in a DNA-dependent and a DNA-independent process. We further found that ICK1/KRP1 misexpression interfered with differentiation and induced cell death, linking cell cycle progression, differentiation, and cell death in plants. The function of ICK1/KRP1 in planta was found to be dependent on a C-terminal domain and regulated negatively by an N-terminal domain. Finally, we identified CDKA;1 and a D-type cyclin as possible targets of ICK1/KRP1 expression in vivo. PMID- 12566575 TI - Quantitative nature of Arabidopsis responses during compatible and incompatible interactions with the bacterial pathogen Pseudomonas syringae. AB - We performed large-scale mRNA expression profiling using an Affymetrix GeneChip to study Arabidopsis responses to the bacterial pathogen Pseudomonas syringae. The interactions were compatible (virulent bacteria) or incompatible (avirulent bacteria), including a nonhost interaction and interactions mediated by two different avirulence gene-resistance (R) gene combinations. Approximately 2000 of the approximately 8000 genes monitored showed reproducible significant expression level changes in at least one of the interactions. Analysis of biological variation suggested that the system behavior of the plant response in an incompatible interaction was robust but that of a compatible interaction was not. A large part of the difference between incompatible and compatible interactions can be explained quantitatively. Despite high similarity between responses mediated by the R genes RPS2 and RPM1 in wild-type plants, RPS2-mediated responses were strongly suppressed by the ndr1 mutation and the NahG transgene, whereas RPM1-mediated responses were not. This finding is consistent with the resistance phenotypes of these plants. We propose a simple quantitative model with a saturating response curve that approximates the overall behavior of this plant-pathogen system. PMID- 12566576 TI - Regulation of actin-dependent cytoplasmic motility by type II phytochrome occurs within seconds in Vallisneria gigantea epidermal cells. AB - The effects of light on actin-dependent cytoplasmic motility in epidermal cells of green leaves of the aquatic angiosperm Vallisneria gigantea were investigated quantitatively using a custom-made dynamic image analyzer. Cytoplasmic motility was measured by monitoring changes in the brightness of individual pixels on digitized images taken sequentially under infrared light. Acceleration and deceleration of cytoplasmic motility were regulated photoreversibly by type II phytochrome(s). This phytochrome-dependent induction of cytoplasmic motility did not occur uniformly in cytoplasm but took place as scattered patches in which no particular organelles, including nucleus, existed. The induction became detectable at 2.5 s after the start of irradiation with pulsed red light. In cells exposed to microbeam irradiation, cytoplasmic motility was induced only in sites in the cytoplasm that were irradiated directly, whereas nonirradiated neighboring areas were unaffected. The effect was short-lived, disappearing within a few minutes, and no signal was transmitted from an irradiated cell to its neighbors. Anti-phytochrome antibody-responsive protein(s) was detectable in the leaf extract by immunoblot and zinc blot analyses and in cryosections of the epidermis by immunocytochemistry. Although the phytochrome-dependent cytoplasmic motility was blocked by exogenously applied latrunculin B or cytochalasins, treatment of the dark-adapted cells with Ca(2+)-chelating reagents induced the cytoplasmic motility. We have proposed a model for the phytochrome regulation of cytoplasmic motility as one of the earliest responses to a light stimulus. PMID- 12566577 TI - The Arabidopsis cax1 mutant exhibits impaired ion homeostasis, development, and hormonal responses and reveals interplay among vacuolar transporters. AB - The Arabidopsis Ca(2+)/H(+) transporter CAX1 (Cation Exchanger1) may be an important regulator of intracellular Ca(2+) levels. Here, we describe the preliminary localization of CAX1 to the tonoplast and the molecular and biochemical characterization of cax1 mutants. We show that these mutants exhibit a 50% reduction in tonoplast Ca(2+)/H(+) antiport activity, a 40% reduction in tonoplast V-type H(+)-translocating ATPase activity, a 36% increase in tonoplast Ca(2+)-ATPase activity, and increased expression of the putative vacuolar Ca(2+)/H(+) antiporters CAX3 and CAX4. Enhanced growth was displayed by the cax1 lines under Mn(2+) and Mg(2+) stress conditions. The mutants exhibited altered plant development, perturbed hormone sensitivities, and altered expression of an auxin-regulated promoter-reporter gene fusion. We propose that CAX1 regulates myriad plant processes and discuss the observed phenotypes with regard to the compensatory alterations in other transporters. PMID- 12566578 TI - HLM1, an essential signaling component in the hypersensitive response, is a member of the cyclic nucleotide-gated channel ion channel family. AB - The hypersensitive response (HR) in plants is a programmed cell death that is commonly associated with disease resistance. A novel mutation in Arabidopsis, hlm1, which causes aberrant regulation of cell death, manifested by a lesion mimic phenotype and an altered HR, segregated as a single recessive allele. Broad spectrum defense mechanisms remained functional or were constitutive in the mutant plants, which also exhibited increased resistance to a virulent strain of Pseudomonas syringae pv tomato. In response to avirulent strains of the same pathogen, the hlm1 mutant showed differential abilities to restrict bacterial growth, depending on the avirulence gene expressed by the pathogen. The HLM1 gene encodes a cyclic nucleotide-gated channel, CNGC4. Preliminary study of the HLM1/CNGC4 gene pro-duct in Xenopus oocytes (inside-out patch-clamp technique) showed that CNGC4 is permeable to both K(+) and Na(+) and is activated by both cGMP and cAMP. HLM1 gene expression is induced in response to pathogen infection and some pathogen-related signals. Thus, HLM1 might constitute a common downstream component of the signaling pathways leading to HR/resistance. PMID- 12566579 TI - The maize genome contains a helitron insertion. AB - The maize mutation sh2-7527 was isolated in a conventional maize breeding program in the 1970s. Although the mutant contains foreign sequences within the gene, the mutation is not attributable to an interchromosomal exchange or to a chromosomal inversion. Hence, the mutation was caused by an insertion. Sequences at the two Sh2 borders have not been scrambled or mutated, suggesting that the insertion is not caused by a catastrophic reshuffling of the maize genome. The insertion is large, at least 12 kb, and is highly repetitive in maize. As judged by hybridization, sorghum contains only one or a few copies of the element, whereas no hybridization was seen to the Arabidopsis genome. The insertion acts from a distance to alter the splicing of the sh2 pre-mRNA. Three distinct intron-bearing maize genes were found in the insertion. Of most significance, the insertion bears striking similarity to the recently described DNA helicase-bearing transposable elements termed HELITRONS: Like Helitrons, the inserted sequence of sh2-7527 is large, lacks terminal repeats, does not duplicate host sequences, and was inserted between a host dinucleotide AT. Like Helitrons, the maize element contains 5' TC and 3' CTRR termini as well as two short palindromic sequences near the 3' terminus that potentially can form a 20-bp hairpin. Although the maize element lacks sequence information for a DNA helicase, it does contain four exons with similarity to a plant DEAD box RNA helicase. A second Helitron insertion was found in the maize genomic database. These data strongly suggest an active Helitron in the present-day maize genome. PMID- 12566580 TI - The beta-subunit of the Arabidopsis G protein negatively regulates auxin-induced cell division and affects multiple developmental processes. AB - Plant cells respond to low concentrations of auxin by cell expansion, and at a slightly higher concentration, these cells divide. Previous work revealed that null mutants of the alpha-subunit of a putative heterotrimeric G protein (GPA1) have reduced cell division. Here, we show that this prototypical G protein complex acts mechanistically by controlling auxin sensitivity toward cell division. Loss-of-function G protein mutants have altered auxin-mediated cell division throughout development, especially during the auxin-induced formation of lateral and adventitious root primordia. Ectopic expression of the wild-type Galpha-subunit phenocopies the Gbeta mutants (auxin hypersensitivity), probably by sequestering the Gbetagamma-subunits, whereas overexpression of Gbeta reduces auxin sensitivity and a constitutively active (Q222L) mutant Galpha behaves like the wild type. These data are consistent with a model in which Gbetagamma acts as a negative regulator of auxin-induced cell division. Accordingly, basal repression of approximately one-third of the identified auxin-regulated genes (47 of 150 upregulated genes among 8300 quantitated) is lost in the Gbeta transcript null mutant. Included among these are genes that encode proteins proposed to control cell division in root primordia formation as well as several novel genes. These results suggest that although auxin-regulated cell division is not coupled directly by a G protein, the Gbeta-subunit attenuates this auxin pathway upstream of the control of mRNA steady state levels. PMID- 12566581 TI - CIPK3, a calcium sensor-associated protein kinase that regulates abscisic acid and cold signal transduction in Arabidopsis. AB - Plants respond to environmental stress by activating "stress genes." The plant hormone abscisic acid (ABA) plays an important role in stress-responsive gene expression. Although Ca(2+) serves as a common second messenger in signaling stress and ABA, little is known about the molecular basis of Ca(2+) action in these pathways. Here, we show that CIPK3, a Ser/Thr protein kinase that associates with a calcineurin B-like calcium sensor, regulates ABA response during seed germination and ABA- and stress-induced gene expression in Arabidopsis. The expression of the CIPK3 gene itself is responsive to ABA and stress conditions, including cold, high salt, wounding, and drought. Disruption of CIPK3 altered the expression pattern of a number of stress gene markers in response to ABA, cold, and high salt. However, drought-induced gene expression was not altered in the cipk3 mutant plants, suggesting that CIPK3 regulates select pathways in response to abiotic stress and ABA. These results identify CIPK3 as a molecular link between stress- and ABA-induced calcium signal and gene expression in plant cells. Because the cold signaling pathway is largely independent of endogenous ABA production, CIPK3 represents a cross-talk "node" between the ABA-dependent and ABA-independent pathways in stress responses. PMID- 12566582 TI - Duplicated fie genes in maize: expression pattern and imprinting suggest distinct functions. AB - Two maize genes with predicted translational similarity to the Arabidopsis FIE (Fertilization-Independent Endosperm) protein, a repressor of endosperm development in the absence of fertilization, were cloned and analyzed. Genomic sequences of fie1 and fie2 show significant homology within coding regions but none within introns or 5' upstream. The fie1 gene is expressed exclusively in the endosperm of developing kernels starting at approximately 6 days after pollination. fie1 is an imprinted gene showing no detectable expression of the paternally derived fie1 allele during kernel development. Conversely, fie2 is expressed in the embryo sac before pollination. After pollination, its expression persists, predominantly in the embryo and at lower levels in the endosperm. The paternal fie2 allele is not expressed early in kernel development, but its transcription is activated at 5 days after pollination. fie2 is likely to be a functional ortholog of the Arabidopsis FIE gene, whereas fie1 has evolved a distinct function. The maize FIE2 and sorghum FIE proteins form a monophyletic group, sharing a closer relationship to each other than to the FIE1 protein, suggesting that maize fie genes originated from two different ancestral genomes. PMID- 12566583 TI - Overexpression of a plasma membrane aquaporin in transgenic tobacco improves plant vigor under favorable growth conditions but not under drought or salt stress. AB - Most of the symplastic water transport in plants occurs via aquaporins, but the extent to which aquaporins contribute to plant water status under favorable growth conditions and abiotic stress is not clear. To address this issue, we constitutively overexpressed the Arabidopsis plasma membrane aquaporin, PIP1b, in transgenic tobacco plants. Under favorable growth conditions, PIP1b overexpression significantly increased plant growth rate, transpiration rate, stomatal density, and photosynthetic efficiency. By contrast, PIP1b overexpression had no beneficial effect under salt stress, whereas during drought stress it had a negative effect, causing faster wilting. Our results suggest that symplastic water transport via plasma membrane aquaporins represents a limiting factor for plant growth and vigor under favorable conditions and that even fully irrigated plants face limited water transportation. By contrast, enhanced symplastic water transport via plasma membrane aquaporins may not have any beneficial effect under salt stress, and it has a deleterious effect during drought stress. PMID- 12566584 TI - Arabidopsis inositol polyphosphate 6-/3-kinase is a nuclear protein that complements a yeast mutant lacking a functional ArgR-Mcm1 transcription complex. AB - Inositol 1,4,5-trisphosphate 3-kinase, and more generally inositol polyphosphate kinases (Ipk), play important roles in signal transduction in animal cells; however, their functions in plant cells remain to be elucidated. Here, we report the molecular cloning of a cDNA (AtIpk2beta) from a higher plant, Arabidopsis. Arabidopsis AtIpk2beta is a 33-kD protein that exhibits weak homology ( approximately 25% identical amino acids) with Ipk proteins from animals and yeast and lacks a calmodulin binding site, as revealed by sequence analysis and calmodulin binding assays. However, recombinant AtIpk2beta phosphorylates inositol 1,4,5-trisphosphate to inositol 1,4,5,6-tetrakisphosphate and also converts it to inositol 1,3,4,5,6-pentakisphosphate [Ins(1,3,4,5,6)P(5)]. AtIpk2beta also phosphorylates inositol 1,3,4,5-tetrakisphosphate to Ins(1,3,4,5,6)P(5). Thus, the enzyme is a D3/D6 dual-specificity inositol phosphate kinase. AtIpk2beta complements a yeast ARG82/IPK2 mutant lacking a functional ArgR-Mcm1 transcription complex. This complex is involved in regulating Arg metabolism-related gene expression and requires inositol polyphosphate kinase activity to function. AtIpk2beta was found to be located predominantly in the nucleus of plant cells, as demonstrated by immunolocalization and fusion to green fluorescent protein. RNA gel blot analysis and promoter-beta-glucuronidase reporter gene studies demonstrated AtIpk2beta gene expression in various organs tested. These data suggest a role for AtIpk2beta as a transcriptional control mediator in plants. PMID- 12566585 TI - Plant gamma-tubulin interacts with alphabeta-tubulin dimers and forms membrane associated complexes. AB - gamma-Tubulin is assumed to participate in microtubule nucleation in acentrosomal plant cells, but the underlying molecular mechanisms are still unknown. Here, we show that gamma-tubulin is present in protein complexes of various sizes and different subcellular locations in Arabidopsis and fava bean. Immunoprecipitation experiments revealed an association of gamma-tubulin with alphabeta-tubulin dimers. gamma-Tubulin cosedimented with microtubules polymerized in vitro and localized along their whole length. Large gamma-tubulin complexes resistant to salt treatment were found to be associated with a high-speed microsomal fraction. Blue native electrophoresis of detergent-solubilized microsomes showed that the molecular mass of the complexes was >1 MD. Large gamma-tubulin complexes were active in microtubule nucleation, but nucleation activity was not observed for the smaller complexes. Punctate gamma-tubulin staining was associated with microtubule arrays, accumulated with short kinetochore microtubules interacting in polar regions with membranes, and localized in the vicinity of nuclei and in the area of cell plate formation. Our results indicate that the association of gamma-tubulin complexes with dynamic membranes might ensure the flexibility of noncentrosomal microtubule nucleation. Moreover, the presence of other molecular forms of gamma-tubulin suggests additional roles for this protein species in microtubule organization. PMID- 12566586 TI - Biosynthesis and emission of terpenoid volatiles from Arabidopsis flowers. AB - Arabidopsis is believed to be mostly self-pollinated, although several lines of genetic and morphological evidence indicate that insect-mediated outcrossing occurs with at least a low frequency in wild populations. Here, we show that Arabidopsis flowers emit both monoterpenes and sesquiterpenes, potential olfactory cues for pollinating insects. Of the 32 terpene synthase genes in the Arabidopsis genome, 20 were found to be expressed in flowers, 6 of these exclusively or almost exclusively so. Two terpene synthase genes expressed exclusively in the flowers and one terpene synthase gene expressed almost exclusively in the flowers were characterized and found to encode proteins that catalyze the formation of major floral volatiles. A beta-glucuronidase fusion construct with a promoter of one of these genes demonstrated that gene expression was restricted to the sepals, stigmas, anther filaments, and receptacles, reaching a peak when the stigma was receptive to cross pollen. The observation that Arabidopsis flowers synthesize and emit volatiles raises intriguing questions about the reproductive behavior of Arabidopsis in the wild and allows detailed investigations of floral volatile biosynthesis and its regulation to be performed with this model plant system. PMID- 12566587 TI - Auxin acts in xylem-associated or medullary cells to mediate apical dominance. AB - A role for auxin in the regulation of shoot branching was described originally in the Thimann and Skoog model, which proposes that apically derived auxin is transported basipetally directly into the axillary buds, where it inhibits their growth. Subsequent observations in several species have shown that auxin does not enter axillary buds directly. We have found similar results in Arabidopsis. Grafting studies indicated that auxin acts in the aerial tissue; hence, the principal site of auxin action is the shoot. To delineate the site of auxin action, the wild-type AXR1 coding sequence, which is required for normal auxin sensitivity, was expressed under the control of several tissue-specific promoters in the auxin-resistant, highly branched axr1-12 mutant background. AXR1 expression in the xylem and interfascicular schlerenchyma was found to restore the mutant branching to wild-type levels in both intact plants and isolated nodes, whereas expression in the phloem did not. Therefore, apically derived auxin can suppress branching by acting in the xylem and interfascicular schlerenchyma, or in a subset of these cells. PMID- 12566588 TI - Role of a single aquaporin isoform in root water uptake. AB - Aquaporins are ubiquitous channel proteins that facilitate the transport of water across cell membranes. Aquaporins show a typically high isoform multiplicity in plants, with 35 homologs in Arabidopsis. The integrated function of plant aquaporins and the function of each individual isoform remain poorly understood. Matrix-assisted laser desorption/ionization time-of-flight analyses suggested that Plasma Membrane Intrinsic Protein2;2 (PIP2;2) is one of the abundantly expressed aquaporin isoforms in Arabidopsis root plasma membranes. Two independent Arabidopsis knockout mutants of PIP2;2 were isolated using a PCR based strategy from a library of plant lines mutagenized by the insertion of Agrobacterium tumefaciens T-DNA. Expression in transgenic Arabidopsis of a PIP2;2 promoter-beta-glucuronidase gene fusion indicated that PIP2;2 is expressed predominantly in roots, with a strong expression in the cortex, endodermis, and stele. The hydraulic conductivity of root cortex cells, as measured with a cell pressure probe, was reduced by 25 to 30% in the two allelic PIP2;2 mutants compared with the wild type. In addition, free exudation measurements revealed a 14% decrease, with respect to wild-type values, in the osmotic hydraulic conductivity of roots excised from the two PIP2;2 mutants. Together, our data provide evidence for the contribution of a single aquaporin gene to root water uptake and identify PIP2;2 as an aquaporin specialized in osmotic fluid transport. PIP2;2 has a close homolog, PIP2;3, showing 96.8% amino acid identity. The phenotype of PIP2;2 mutants demonstrates that, despite their high homology and isoform multiplicity, plant aquaporins have evolved with nonredundant functions. PMID- 12566589 TI - The biosynthesis of L-arabinose in plants: molecular cloning and characterization of a Golgi-localized UDP-D-xylose 4-epimerase encoded by the MUR4 gene of Arabidopsis. AB - The mur4 mutant of Arabidopsis shows a 50% reduction in the monosaccharide L-Ara in leaf-derived cell wall material because of a partial defect in the 4 epimerization of UDP-D-Xyl to UDP-L-Ara. To determine the genetic lesion underlying the mur4 phenotype, the MUR4 gene was cloned by a map-based procedure and found to encode a type-II membrane protein with sequence similarity to UDP-D Glc 4-epimerases. Enzyme assays of MUR4 protein expressed in the methylotropic yeast Pichia pastoris indicate that it catalyzes the 4-epimerization of UDP-D-Xyl to UDP-L-Ara, the nucleotide sugar used by glycosyltransferases in the arabinosylation of cell wall polysaccharides and wall-resident proteoglycans. Expression of MUR4-green fluorescent protein constructs in Arabidopsis revealed localization patterns consistent with targeting to the Golgi, suggesting that the MUR4 protein colocalizes with glycosyltransferases in the biosynthesis of arabinosylated cell wall components. The Arabidopsis genome encodes three putative proteins with >76% sequence identity to MUR4, which may explain why mur4 plants are not entirely deficient in the de novo synthesis of UDP-L-Ara. PMID- 12566590 TI - The roles of auxin response factor domains in auxin-responsive transcription. AB - Auxin response factors (ARFs) are transcription factors that bind to TGTCTC auxin response elements in promoters of early auxin response genes. ARFs have a conserved N-terminal DNA binding domain (DBD) and in most cases a conserved C terminal dimerization domain (CTD). The ARF CTD is related in amino acid sequence to motifs III and IV found in Aux/IAA proteins. Just C terminal to the DBD, ARFs contain a nonconserved region referred to as the middle region (MR), which has been proposed to function as a transcriptional repression or activation domain. Results with transfected protoplasts reported here show that ARFs with Q-rich MRs function as activators, whereas most, if not all other ARFs, function as repressors. ARF DBDs alone are sufficient to recruit ARFs to their DNA target sites, and auxin does not influence this recruitment. ARF MRs alone function as activation or repression domains when targeted to reporter genes via a yeast Gal4 DBD, and auxin does not influence the potency of activation or repression. ARF CTDs, along with a Q-rich MR, are required for an auxin response whether the MRs plus CTDs are recruited to a promoter by an ARF DBD or by a Gal4 DBD. The auxin response is mediated by the recruitment of Aux/IAA proteins to promoters that contain a DNA binding protein with a Q-rich MR and an attached CTD. PMID- 12566591 TI - The eto1, eto2, and eto3 mutations and cytokinin treatment increase ethylene biosynthesis in Arabidopsis by increasing the stability of ACS protein. AB - The Arabidopsis ethylene-overproducing mutants eto1, eto2, and eto3 have been suggested to affect the post-transcriptional regulation of 1-aminocyclopropane-1 carboxylic acid synthase (ACS). Here, we present the positional cloning of the gene corresponding to the dominant eto3 mutation and show that the eto3 phenotype is the result of a missense mutation within the C-terminal domain of ACS9, which encodes one isoform of the Arabidopsis ACS gene family. This mutation is analogous to the dominant eto2 mutation that affects the C-terminal domain of the highly similar ACS5. Analysis of purified recombinant ACS5 and epitope-tagged ACS5 in transgenic Arabidopsis revealed that eto2 does not increase the specific activity of the enzyme either in vitro or in vivo; rather, it increases the half life of the protein. In a similar manner, cytokinin treatment increased the stability of ACS5 by a mechanism that is at least partially independent of the eto2 mutation. The eto1 mutation was found to act by increasing the function of ACS5 by stabilizing this protein. These results suggest that an important mechanism by which ethylene biosynthesis is controlled is the regulation of the stability of ACS, mediated at least in part through the C-terminal domain. PMID- 12566593 TI - Dopaminergic modulation of visual-spatial working memory in Parkinson's disease. AB - Visual-spatial working memory (WM) impairment is frequently associated with the early stage of Parkinson's disease (PD). The aim of this study was to evaluate the performance of a group of PD patients in visual-spatial and visual-object WM tasks and to investigate the effect of administering the dopaminergic agonist apomorphine (experiment 1) or the dopamine precursor L-dopa (experiment 2) on the performance of tests assessing these functions. To study WM processes, the PD patients and age-matched normal controls were given an n-back task paradigm. In both experiments, the PD patients were submitted to two evaluations: one after a 12-hour therapy washout and the other 15 min after a subcutaneous infusion of apomorphine (average 0.04 mg/kg) or 20/30 min after L-dopa intake (200 mg p.o.). The apomorphine infusion had a worsening effect on reaction times in both visual spatial and visual-object WM tasks, but it did not influence performance accuracy. Instead, L-dopa administration had a ameliorative effect on accuracy and reaction times in both visual-spatial and visual-object tasks. These results highlight the role of dopamine in the modulation of the WM function in PD patients. PMID- 12566592 TI - Genomic analysis of the unfolded protein response in Arabidopsis shows its connection to important cellular processes. AB - We analyzed the breadth of the unfolded protein response (UPR) in Arabidopsis using gene expression analysis with Affymetrix GeneChips. With tunicamycin and DTT as endoplasmic reticulum (ER) stress-inducing agents, we identified sets of UPR genes that were induced or repressed by both stresses. The proteins encoded by most of the upregulated genes function as part of the secretory system and comprise chaperones, vesicle transport proteins, and ER-associated degradation proteins. Most of the downregulated genes encode extracellular proteins. Therefore, the UPR may constitute a triple effort by the cell: to improve protein folding and transport, to degrade unwanted proteins, and to allow fewer secretory proteins to enter the ER. No single consensus response element was found in the promoters of the 53 UPR upregulated genes, but half of the genes contained response elements also found in mammalian UPR regulated genes. These elements are enriched from 4.5- to 15-fold in this upregulated gene set. PMID- 12566594 TI - Rates of cognitive decline in Alzheimer's disease and dementia with Lewy bodies. AB - Increased interest in types of dementia has developed as more cases are identified in aging populations. Here we compare the rates of cognitive decline over time in three groups with dementia from the University of Western Ontario Dementia Study: Alzheimer's disease (AD), dementia with Lewy bodies and a group with both AD and Lewy bodies. All diagnoses were verified by autopsy using standard diagnostic methods. Cognitive impairment was measured with the Extended Scale for Dementia (ESD). Members of each group with dementia were age and sex matched with individuals without dementia as controls. The 15 cases of AD, 7 cases with Lewy bodies and 8 cases with both conditions were all free of significant vascular disease. Linear regression was used to determine the rate of changes in ESD scores over time in months. All three control groups showed no change in cognitive status over time. As expected, all groups with dementia showed progressive cognitive impairment. Analysis of the slope parameter showed that all groups deteriorated at the same rate of approximately 2 ESD points per month. Quadratic models fit better than simple linear models in all groups. Results suggest that the final rate of cognitive decline in dementia may not necessarily reflect the underlying cause. PMID- 12566595 TI - Synapse loss may be a minor contributor to decreased regional cerebral blood flow in Alzheimer disease. AB - In this study we tested the hypothesis that synapse loss contributes to decreased regional cerebral blood flow (rCBF) in Alzheimer disease (AD). We compared antemortem rCBF and postmortem analysis of synaptophysin, as a measure of synapse loss, in 13 cases of AD. rCBF studies were performed using inhaled xenon gas (Xe 133), which yields quantitative results. Synapse loss was evaluated in postmortem brain samples using an enzyme-linked immunosorbant assay (ELISA) that measures synaptophysin, with results expressed as picomoles synaptophysin/10 mg brain. Synaptophysin was expressed either as concentration (QS method) or as the ratio of the concentration to the combined results in frontal, temporal and parietal lobe (RS method). There was no correlation between synapse loss and rCBF using the QS method and only borderline significance between right SPECT and right temporal synaptophysin using the RS method. The results of this study suggest that synapse loss may be a minor contributor to the decreased rCBF observed in AD. PMID- 12566596 TI - Galantamine provides sustained benefits in patients with 'advanced moderate' Alzheimer's disease for at least 12 months. AB - Galantamine (Reminyl), a novel agent with a dual mode of action, modulates nicotinic acetylcholine receptors and inhibits acetylcholinesterase. Galantamine has consistently demonstrated a broad range of beneficial effects and has shown sustained benefits in cognitive and functional abilities for at least 12 months in patients with mild-to-moderate Alzheimer's disease (AD). As pivotal studies demonstrating the efficacy of cholinergic drugs were designed to exclude patients with severer AD, many patients with the advanced stage of this condition are currently not treated due to the lack of demonstrated efficacy in clinical trials. We aimed to investigate whether there was any evidence for the benefits of galantamine in patients with severer disease, by performing a post hoc analysis using data extracted from the population of the two long-term galantamine studies. We evaluated the efficacy of galantamine in patients with 'advanced moderate' AD. 'Advanced moderate' patients were those with baseline Mini Mental State Examination (MMSE) scores 30. These patients were compared with matched controls who received placebo in a different historical study. Cognitive abilities (assessed using the ADAS-cog scale) of 'advanced moderate' AD patients receiving galantamine for 12 months were maintained at baseline levels after 12 months, and significantly improved over those of placebo patients (p < 0.001). Of the 'advanced moderate' patients receiving galantamine, 51% with baseline ADAS-cog of >30 maintained or improved their ADAS-cog scores over baseline values, compared with 13% receiving placebo (p < 0.001). In the subgroup of 'advanced moderate' patients with baseline MMSE 60% of the hospitals have established ECs. Sixty-five percent of the responders believe that the EC is an advisory committee and 31% an executive committee. Endoscopists believe that ECs have been of great help to their specialty in their country. CONCLUSIONS: The roles of the ECs are not clear to endoscopists and there is a need for education in this field. PMID- 12566613 TI - Chromoendoscopy: from a research tool to clinical progress. AB - Chromoendoscopy, the intravital staining of gastrointestinal epithelia, provides additional diagnostic information with respect to the epithelial morphology and pathophysiology. Based on experience gathered mainly in Japan, chromoendoscopy is now in more widespread use, in particular to identify preneoplastic and neoplastic lesions. The most promising techniques are the depiction of squamous epithelium neoplasia of the esophagus with Lugol's solution, staining of Barrett's mucosa by methylene blue, including the potential to identify neoplasia, and the demarcation of neoplasia with indigo carmine in stomach and colon for local endoscopic resection. However, the optimal methodology is still to be defined as well as diverging results of diagnostic accuracy to be clarified. High-resolution and magnifying endoscopy have breathed new life into chromoendoscopy. Innovative applications and refinement of the existing ones are soon to be expected. PMID- 12566614 TI - Endoscopic ultrasound in pancreatic diseases. AB - There are many indications for the use of endoscopic ultrasound (EUS) in the management of patients with pancreatic diseases. High-resolution imaging of the pancreas is achievable due to the close proximity between luminal structures and the pancreas. Since its introduction, EUS has had a significant impact on the diagnosis of pancreatic diseases. The detection of small lesions and neuroendocrine pancreatic tumors as well as the preoperative staging of pancreatic adenocarcinoma have been improved employing EUS. For the detection of small pancreatic tumors <2 cm in diameter, EUS appears to be the most sensitive method. EUS adds significant information to the differential diagnosis between pancreatic cancer and chronic pancreatitis, and it may be further enhanced by EUS guided fine-needle aspiration. While the role of EUS in distinguishing between benign and malignant cystic pancreatic tumors is under discussion, EUS-guided drainage of pancreatic pseudocysts is an accepted treatment option for symptomatic individuals. One of the most important advantages of EUS apart from tumor staging is the early detection of chronic pancreatitis. EUS is as good as endoscopic retrograde cholangiopancreatography in diagnosing chronic pancreatitis in advanced stages. In early stages of the disease, when the ductal system remains normal, EUS appears to be a superior diagnostic modality because it can detect features of chronic pancreatitis in the parenchyma not visible by other techniques. PMID- 12566615 TI - The wireless capsule: new light in the darkness. AB - The development of wireless capsule endoscopy offers the potential to examine the whole small intestine, with its 5-meter length, and has the advantage of being painless. Using a miniature CMOS camera and a short focal length lens, images are obtained, as the optical window of the capsule sweeps past the gut wall, without requiring air inflation of the gut lumen. The capsule endoscope is propelled by peristalsis through the gastrointestinal tract. The video images are transmitted using radiotelemetry to an array of aerials attached to the body which allows image capture. The images are stored on a small portable recorder carried on a belt and subsequently downloaded for analysis. The system allows more than 7 h of continuous recording of images of the gastrointestinal tract. The patients are free to continue their daily routine during the examination. The capsule endoscope has performed well in trials in patients with difficult gastrointestinal bleeding and in comparative studies with push enteroscopy. It has received both a CE mark and FDA approval for use in patients. It has been used to date in about 4,000 patients. The current clinical data are reviewed. PMID- 12566616 TI - Update on treatment of variceal hemorrhage. AB - Variceal hemorrhage accounts for one third of all deaths related to cirrhosis. To date, many modalities of treating variceal bleeding have been devised, including pharmacological therapy. Treatment of variceal hemorrhage includes resuscitation, initial hemostasis, and prevention of complications and recurrent bleeding. Intravenous vasoactive agents such as terlipressin, somatostatin, octreotide, or vapreotide should be administered in patients with suspected variceal bleeding. Endoscopic treatment remains the mainstay of treatment. Endoscopic variceal ligation is safer and more efficacious than sclerotherapy as initial treatment of bleeding esophageal varices, whereas cyanoacrylate injection is the endoscopic treatment of choice for gastric varices. An adjuvant vasoactive agent is useful for the prevention of early rebleeding. Prophylactic antibiotics are increasingly used for prevention of infection, notably spontaneous bacterial peritonitis. Follow-up endoscopic treatment is necessary in order to obliterate residual varices. The combination of a beta blocker and nitrate is an essential component of secondary prophylaxis for recurrent variceal bleeding. Transjugular intrahepatic portosystemic shunt or surgery offers the best salvage therapy in patients with failed hemostasis or breakthrough recurrent bleeding despite medical and endoscopic therapy. Endoscopic ultrasonography is useful in the prediction of recurrence of varices and facilitates visualization and guidance of further treatment of gastric varices. Despite advances in the treatment of variceal bleeding, liver function remains the determining factor of patient survival. Liver transplantation is the only definitive treatment that can alter the course of the disease. PMID- 12566617 TI - Endoscopic techniques for enteral nutrition: standards and innovations. AB - Enteral nutrition has been increasingly used in clinical practice during the past decades. Today, nasogastric, nasoenteric, and transcutaneous gastric or enteral feeding tubes are well established as a routine endoscopic intervention. After clinical introduction of percutaneous endoscopic gastrostomy (PEG) in 1980, the sutureless 'pull' PEG has become a widespread endoscopic technique for transcutaneous gastric long-term nutrition. Multiple new techniques have been introduced since then, and today even long-term jejunal nutrition can be achieved with modified techniques. The introducer PEG (first reported in 1984) has not become an established procedure, but new techniques with an endoscopic gastropexy might be a more effective approach. To increase the quality of life of the patients, skin-level devices were designed and successfully introduced in 1984. A new development has been the one-step feeding tubes which provide the patients with a permanent tube. The standard techniques for long-term enteral feeding, new developments of feeding tubes, and future concepts are discussed in this review. PMID- 12566618 TI - Stenting in esophageal strictures. AB - The interventional management of esophageal strictures remains, to date, an important clinical challenge. Stenting is probably the best palliation modality in patients with incurable esophagogastric carcinoma. Conversely, the use of esophageal stents is still relatively uncommon for the treatment of refractory benign strictures. In the last few years, several new stents have become available as a result of significant advances that have been made in terms of design and materials. This review focuses on the endoscopic use of esophageal stents in malignant and benign esophageal strictures, revisiting the different types of expandable stents presently available, the techniques, the results, and the complications of stent insertion and giving some practical advices. Future developments in the field of esophageal stenting are also discussed. PMID- 12566620 TI - Stenting of the gastrointestinal tract. AB - Self-expandable metal stents are being increasingly used to palliate malignant stenoses of the gastrointestinal tract. These prostheses have made it possible to reestablish luminal continuity in patients with malignant obstruction of esophagus, gastric outlet, small bowel, or colon who are at high risk of surgical intervention. Despite technologic refinements in the stents and their delivery systems, and an increasing number of studies related to self-expandable metal stents, data regarding their long-term efficacy, side effect profile, and true cost efficacy in comparison with other treatment modalities remain incomplete. This article reviews the available technology, placement techniques, indications, and outcomes relating to the use of self-expandable metal stents in esophagus, stomach, small bowel, and colon. PMID- 12566619 TI - Endoscopic interventions for preneoplastic and neoplastic lesions: mucosectomy, argon plasma coagulation, and photodynamic therapy. AB - The classical treatment of neoplastic lesions is surgical resection. Improved detection of early lesions has offered new therapeutic strategies. Thus, for the last 20 years it has been increasingly attempted to undertake endoscopic resection of such tumors. While there is no doubt that morbidity and mortality are lower than after surgery, it was assumed that the long-term prognosis is not as good as with surgery. Surprisingly, however, the 5-year survival rates are excellent. Therefore, in local mucosal lesions of colon, stomach, and esophagus, endoscopic removal has become standard treatment. In Barrett's esophagus, the treatment modality it is still discussed controversially, despite encouraging results. Since we are yet not able to completely eliminate the premalignant specialized intestinal epithelium of the esophagus, it is still uncertain whether eradication of the malignant or dysplastic lesion is sufficient. PMID- 12566621 TI - Innovations in gastro-intestinal endoscopy: endoscopic antireflux therapies for gastro-oesophageal reflux disease. AB - BACKGROUND: Gastro-oesophageal reflux disease (GORD) is a common disorder which significantly impairs the quality of life. Recently a number of endoscopic antireflux procedures have been described, with a view to the management of GORD, thus obviating the need for long-term proton pump inhibitors and the cost and potential risk of laparoscopic Nissen fundoplication. AIM: To evaluate the status of new techniques using gastro-intestinal endoscopy for the management of GORD. METHODS: Review of the literature (publications and abstracts) regarding safety, efficacy, and durability of these new antireflux procedures. RESULTS: These antireflux techniques have produced significant improvement in GORD symptomatology and quality of life and have reduced the use of antireflux medication, without causing serious morbidity or mortality. However, the majority of these techniques have failed to adequately control oesophageal acid reflux. CONCLUSIONS: Endoscopic antireflux therapies have the potential to treat patients with GORD. However, further studies are necessary to determine what modifications to these techniques are required in order to produce the maximum clinical efficacy and durability. PMID- 12566622 TI - Self-administered symptom questionnaires in patients with dyspepsia and their yield in discriminating between endoscopic diagnoses. AB - BACKGROUND/AIMS: Symptoms are generally considered to be poor predictors of organic findings in patients with dyspepsia. We aimed at evaluating whether specific gastrointestinal symptoms, identified by self-administered questionnaires, correlate with specific endoscopic diagnoses and discriminate organic from functional dyspepsia. METHODS: Adult patients with pain or discomfort centred in the upper abdominal region were consecutively enrolled. Patients with heartburn, acid regurgitation, or defaecation and bowel habit problems as their predominant symptoms were excluded. Three self-administered questionnaires were applied before an oesophagogastroduodenoscopy was performed. RESULTS: Among the 799 patients, 50.6% had a normal endoscopy. Endoscopic diagnoses comprised: non-erosive oesophagitis (7.5%), erosive oesophagitis (11.1%), Barrett's oesophagus (1.1%), gastritis/duodenitis (8.4%), gastric ulcer (4.5%), duodenal ulcer (8.3%), and cancer (1.3%). Non-dominant heartburn and acid regurgitation were significantly more common in patients with organic dyspepsia, whereas hunger pains and rumbling occurred more often in those with functional dyspepsia. Multivariate analyses demonstrated that younger age, female gender, high scores for hunger pain, rumbling, hard stools, low scores for heartburn, and acid regurgitation predicted functional dyspepsia. CONCLUSIONS: Self-administered questionnaires revealed differences in the symptom patterns between patients with functional and organic dyspepsia. Furthermore, the health-related well-being in patients with functional and organic dyspepsia centred was impaired to the same extent. PMID- 12566623 TI - Treatment of benign biliary strictures in chronic pancreatitis by self-expandable metal stents. AB - AIM: To determine the efficacy the value of self-expandable metal stents in patients with benign biliary strictures caused by chronic pancreatitis. METHOD: 61 patients with symptomatic common bile duct strictures caused by alcoholic chronic pancreatitis were treated by interventional endoscopy. RESULTS: Initial endoscopic drainage was successful in all cases, with complete resolution of obstructive jaundice. Of 45 patients who needed definitive therapy after a 12 months interval of interventional endoscopy, 12 patients were treated with repeated plastic stent insertion (19.7%) or by surgery (n = 30; 49.2%). In 3 patients a self-expandable metal stent was inserted into the common bile duct (4.9%). In patients treated with metal stents, no symptoms of biliary obstruction occurred during a mean follow-up period of 37 (range 18-53) months. The long-term success rate of treatment with metal stents was 100%. CONCLUSIONS: Endoscopic drainage of biliary obstruction by self-expandable metal stents provides excellent long-term results. To identify patients who benefit most from self expandable metal stent insertion, further, prospective randomized studies are necessary. PMID- 12566624 TI - The prechordal plate, the rostral end of the notochord and nearby median features in staged human embryos. AB - The enigmatic structure known as the prechordal plate and also the precursors of the notochord were reassessed in 101 human embryos of stages 8-14; 36 were controlled by precise graphic reconstructions. Various measurements were made and the appearance of median structures was tabulated. The prechordal plate, which has been unequivocally found first at stage 7, is usually detectable at stage 8 as a highly developed mesendodermal mass in contact with the floor of the neural groove. At stages 9 and 10 the plate is related to neuromere D1. Cellular migration laterad at stages 9-11 gives rise to the bilateral premandibular condensations, which are lateral to the adenohypophysial primordium, and at stages 13 and 14 these condensations are closely related to the future tentorium cerebelli. The notochordal process is first visible at stage 7, and its dorsal part constitutes the notochordal plate at stage 8. At stages 8-10 the notochordal and prechordal plates appear continuous, but they are distinguishable histologically. The notochordal plate becomes intercalated in the endoderm of the foregut and begins to give rise to the notochord at stages 10 and 11. Bifurcation occurs rostrally at stage 12: the dorsal limb disappears, whereas the ventral limb is the definitive continuation. The topographical relationships of the prechordal and notochordal plates, the notochord, the adenohypophysis, and the oropharyngeal membrane are documented. Definitions and pertinent remarks on terminology are included, comparative data are considered, and the origin and derivatives of the prechordal plate are discussed. In addition to giving rise to external ocular muscles, the possibility of contributions to the heart and the tentorium cerebelli is raised. The importance of the plate in the development of the forebrain, as well as in the production of median anomalies such as holoprosencephaly and cyclopia, is stressed. PMID- 12566625 TI - Removal of the shell coat affects maintenance of epithelia in blastocysts of the Brushtail Possum in vitro. AB - The aim of the present study was to investigate the role of the marsupial shell coat in embryonic development because it may be a potential target for immunocontraceptive control of the brushtail possum. Conceptuses from 52 female possums were collected between 1995 and 1997 in New Zealand and Australia. Development was examined in representative stages from cleavage to the early trilaminar blastocyst. The effect of coat removal by microdissection was examined by comparing development in vivo (n = 29), with development in vitro, both with (n = 10) and without (n = 13) shell coat removal. Conceptuses were monitored and photographed in culture, fixed and examined by transmission electron microscopy. The ultrastructure of uni-, bi- and trilaminar blastocyst stages in vivo and in vitro and of the early stages of hypoblast and embryonic endoderm formation are described for the first time in the possum. Shell coat removal had little impact on most cleavage stages, as the intact mucoid coat appeared to provide structural support to the embryo. Common features of unilaminar coat-free specimens after culture were rounding up and detachment of cells from the blastocyst epithelium and the loss of cell surface projections. The most remarkable features of the shell-free trilaminar blastocysts in comparison with the in vivo and in vitro controls were the hydration of many cells, and the large-scale disruption and modification of numerous epithelia, particularly of the younger, or newly forming populations. The shell appears to be functionally important after blastocyst formation, particularly after breakdown of the mucoid coat. After shell removal, conceptus response in vitro suggested that the shell played a role in maintaining structural integrity. The shell was found to be necessary for normal embryonic development. PMID- 12566626 TI - Cyclic changes of the canine endometrial surface: an electron-microscopic study. AB - The endometrial surface morphology of 38 dogs during different stages of the estrous cycle was investigated with scanning electron microscopy. The cell surface altered from convex in proestrus and estrus to very variable in early metestrus, flattened in late metestrus and became completely plane in anestrus. Microvilli were numerous and long in proestrus and in estrus, became short and variable in number in early metestrus, decreased further in length in late metestrus and became very short and rare in anestrus. The variable appearance in early metestrus was not influenced by changing the osmolarity of the fixative and might be a physiological process. The number of glandular openings showed little variability throughout the estrous cycle. Ciliated cells were rare but present in all cycle stages except in late metestrus. However, in the latter cycle stage and in anestrus rare single strands were noted. Transmission electron microscopy was used to determine the inner structure of these strands. Microtubuli were detected in transversal and longitudinal sections but without the 9 + 2 arrangement which is characteristic for cilia. The nature and function of these structures remain unclear. PMID- 12566627 TI - Sarcoglycans in human skeletal muscle and human cardiac muscle: a confocal laser scanning microscope study. AB - Sarcoglycans are a subcomplex of transmembrane proteins which are part of the dystrophin-glycoprotein complex. They are expressed in the skeletal, cardiac and smooth muscle. Although numerous studies have been conducted on the sarcoglycan subcomplex in skeletal and cardiac muscle, the manner of the distribution and localization of these proteins along the nonjunctional sarcolemma is not clear. We therefore carried out an indirect immunofluorescence study on surgical biopsies of normal human skeletal muscle and of healthy human atrial myocardium biopsies of patients affected by valvulopathy. Our results indicate that, in skeletal muscle, sarcoglycans have a costameric distribution and all colocalize with each other. Only in a few cases did the alpha-sarcoglycan not colocalize with other sarcoglycans. In addition, these glycoproteins can be localized in different fibers either in the regions of the sarcolemma over band I or band A. In cardiac muscle, our results show a costameric distribution of all proteins examined and, unlike in skeletal muscle, they show a constant colocalization of all sarcoglycans with each other, along with a consistent localization of these proteins in the region of the sarcolemma over band I. In our opinion, this situation seems to confirm the hypothesis of a correlation between the region of the sarcolemma occupied by costameric proteins and the metabolic type, fast or slow, of the muscular fibers. These data, besides opening a new line of research in understanding interactions between the sarcoglycans and other transmembrane proteins, could also be extended to skeletal and cardiac muscles affected by neuromuscular and cardiovascular pathologies to understand possible structural alterations. PMID- 12566628 TI - Ultrastructural identification of P2Y2 receptor mRNA in the rat thymus. AB - In situ hybridisation to identify P2Y(2) receptor mRNA was performed for the first time at the ultrastructural level on the thymus of adult male rats. These studies revealed transcripts for P2Y(2) receptors in cortical T cells and endothelial cells of thymic blood vessels. These transcripts are likely to be linked with the production of functional P2Y(2) receptors in these cells. In the T cells, transcripts for the P2Y(2 )receptor were localised in the cytoplasm as well as on the smooth endoplasmic reticulum and cell membrane. Dividing T cells also expressed P2Y(2 )receptor mRNA, mostly in the cytoplasm around chromosomal material. Endothelial cells displaying labels for P2Y(2 )receptor transcripts were of cortical arteries/arterioles and capillaries and of postcapillary venules in the corticomedullary junction. P2Y(2) mRNA transcripts were localised in the cytoplasm of endothelial cells, although they did not appear to be specifically associated with subcellular organelles or structures. In postcapillary venules, T cells displaying labelling for the P2Y(2) receptor were seen migrating across the P2Y(2) receptor mRNA-positive endothelium. Our findings are discussed in terms of the relationship between thymic immune cells and the endothelium. This includes the issue of immune cell trafficking into the circulation, and the ATP-related regulatory role and involvement of P2Y(2) receptors in the rat thymus. PMID- 12566629 TI - Collagenase-induced changes in articular cartilage as detected by electron microscopic stereology, quantitative polarized light microscopy and biochemical assays. AB - The purpose of this study was to compare the ability of electron-microscopic (EM) stereology with quantitative polarized light microscopy (PLM) and biochemical collagen (hydroxyproline) and crosslink (pyridinoline) analyses to detect changes in the superficial collagen network of bovine articular cartilage after digestion in vitro with purified bacterial (Clostridium histolyticum) collagenase (30 U/ml) for 24 and 48 h. Collagen volume (V(V)) and surface (S(V)) densities of the uppermost third of the superficial zone were estimated indirectly from zonal isotropic uniform random sections using collagen length density (L(V)) and average collagen fibril diameter, or its average second power. Collagenase digestion caused a significant decrease in fibril diameter (64 to 62%), V(V) (89 to 95%) and S(V) (64 to 86%) after incubation for 24 and 48 h. Collagen L(V) remained unchanged after 24 h incubation but decreased 63% after 48 h. Collagen concentration per dry weight, assayed biochemically from the whole superficial zone, decreased also significantly (29 to 60%) after 24 and 48 h digestions, respectively. The pyridinoline concentration per dry weight of the superficial zone decreased (31 to 57%) whereas the pyridinoline concentration per collagen remained unchanged. PLM revealed that the birefringence of the uppermost third of the superficial zone was decreased by 36% after digestion for 24 h though the total birefringence of the whole zone was not reduced. However, after 48 h, the birefringence of the whole superficial zone was significantly reduced (76%). All of the techniques compared in this study could detect collagen network degradation in bovine articular cartilage but the EM stereological technique was more sensitive at detecting the changes than PLM or biochemical assays. PMID- 12566630 TI - Changes in the epithelium of the vaginal complex during the estrous cycle of the marsupial Monodelphis domestica. 1. Transmission electron microscopy study. AB - The vaginal complex of marsupials differs from that of eutherians. Cervices open separately in a sinus vaginalis or cul-de-sac. Two lateral vaginae adjoin the sinus vaginalis and fuse at the level of the urethra opening and form the sinus urogenitalis. During the estrous cycle the vaginal epithelium undergoes a number of specified morphological changes. This paper is the first to describe these changes on an ultrastructural level in a marsupial. Investigations in Monodelphis vagina reveal that a cyclic switch exists between a keratinized and a stratified nonkeratinized epithelium. Keratinization starts during proestrus and reaches its maximum during estrus. In the postestrus, desquamation of the stratum corneum takes place, mostly in two steps. In metestrus one to two additional layers of the now nonkeratinized surface cells are shed into the vaginal lumen. Typical cell structures, such as keratin filaments, keratohyalin and membrane-coating granules, are involved in the keratinization process. Keratohyalin is found in the cytoplasm as well as in the nucleus of stratum granulosum cells, a phenomenon which is known from other parakeratinized epithelia of rapid turnover. Membrane coating granules, responsible for the permeability barrier between the epithelial cells, are of the nonlamellated type in the nonkeratinized epithelium and produce an amorphous material in the intercellular spaces after extrusion. At periods, however, when the epithelium is keratinized, membrane-coating granules are of the lamellated type and form a lamellated barrier structure after extrusion in the intercellular space. The loss of the protective keratinized layers asks for an additional defense mechanism for the epithelium. The migration of leukocytes through the epithelium predominantly during post- and metestrus and their presence in the vaginal lumen may play a protective role together with the bacterial content. PMID- 12566631 TI - Morphology of the intrinsic cardiac nervous system in the dog: a whole-mount study employing histochemical staining with acetylcholinesterase. AB - The intrinsic cardiac nervous system is known to be important both in the prevention and treatment of risky heart diseases. The present study was designed to determine the topography and 3-dimensional architecture of the intrinsic nervous system in the canine heart highlighting the differences of this system in dogs and humans. The morphology of the intrinsic cardiac neural plexus was revealed with a histochemical method using acetylcholinesterase in whole hearts of 18 mongrel dogs and examined with the aid of dissecting stereoscopic and contact microscopes. The present study identified 13 locations between the canine ascending aorta and pulmonary trunk, around the pulmonary veins, and on every side of the superior vena cava, through which mediastinal cardiac nerves accessed the canine heart. Intrinsic nerves from these locations extended within the canine epicardium by seven neuronal subplexuses. Intrinsic nerves and ganglia were found to be widely distributed in topographically consistent atrial and ventricular regions. In general, the canine right atrium, including the sinoatrial node, was innervated by two subplexuses, the wall of the left atrium by three, and the right and left ventricles by two subplexuses. Depending on the age of the animal, the number of intrinsic ganglia per one canine heart might range from 400 up to 1,500. By taking into account the ganglion size and potential approximate number of neurons residing inside a ganglion of a certain size, it was estimated that on average about 80,000 intrinsic neurons are associated with the canine heart. A comparative analysis of the morphological patterns of the canine and human intrinsic cardiac neural plexuses showed that the topographies of these plexuses may be considered as quite similar, but the structural and quantitative differences of the intrinsic cardiac neural subplexuses between dogs and humans are significant. PMID- 12566632 TI - Performance of conventional and new methods for the detection of occlusal caries in deciduous teeth. AB - The aim of this study was to compare, in vitro, the performance of different conventional diagnostic techniques with the laser-based device, DIAGNOdent, for diagnosis of occlusal caries in deciduous teeth. 95 deciduous teeth with macroscopically intact occlusal surfaces were selected. All teeth were assessed by the following techniques: visual inspection (VI), visual inspection with magnification (VIM), visual inspection combined with light pressure probing (VIP), bitewing radiography (BW) and the DIAGNOdent (DD). Caries extension was assessed by histology. DIAGNOdent showed, in comparison with the conventional clinical methods, a significantly improved ability (p < 0.05) to detect dentinal lesions (D(3) or D(4)) in deciduous teeth and an overall performance similar to that obtained in previous studies on permanent teeth. There was no statistically significant difference between VIM and DD in detecting caries confined to enamel (D(2)). DD performed better at D(2) level than the other methods. The intraexaminer kappa scores for DIAGNOdent were 0.76-0.86 (D(2)) and 0.77-0.85 (D(3)), respectively. It was concluded that DIAGNOdent could be used as an additional tool in the detection of occlusal caries in deciduous teeth and its good reproducibility should enable the laser device to monitor the caries process over time. PMID- 12566633 TI - An in vitro comparison of a combined FOTI/visual examination of occlusal caries with other caries diagnostic methods and the effect of stain on their diagnostic performance. AB - The aims of this in vitro study were to compare the combination of FOTI and visual inspection to other diagnostic methods for the detection and evaluation of the depth of occlusal caries and assess the effect of stain on diagnostic performance. Five diagnostic methods (combined FOTI/Visual (CFV), Visual, FOTI, DIAGNOdent and Electrical Caries Monitor) were considered. Histological validation was performed in the 152 occlusal molar sites (111 teeth) using 350 microm sections and a stereomicroscope. The rank correlation coefficients with histology ranged from 0.42 (DIAGNOdent) to 0.66 (CFV). The area under the ROC curves (AUC) for enamel lesions ranged from 0.82 (ECM) to 0.88 (CFV) and from 0.81 (DIAGNOdent) to 0.91 (CFV) for dentine lesions. The AUC at the dentine level for CFV and FOTI were similar (p > 0.05), but the AUC for CFV was significantly greater than for visual (p < 0.001), DIAGNOdent (p = 0.005) and the ECM (p = 0.04). FOTI was found to be particularly useful for the detection of dentine lesions. The exclusion of stain and brown spot lesions improved performance for all methods. It is concluded that CFV is useful for the determination of occlusal lesion depth and that in the presence of stain and brown spot lesions different cut-off points may be required for the ECM and DIAGNOdent to identify dentine lesions. PMID- 12566635 TI - Influence of enamel thickness on quantification of mineral loss in enamel using laser-induced fluorescence. AB - The objective of this study was to evaluate the effect of enamel thickness on the quantification of white-spot lesions with laser fluorescence. One hundred and twenty 3-mm-diameter human ground and polished enamel specimens were used. Specimens were illuminated by a 488-nm argon laser, images were acquired through a 540-nm high-pass filter and stored on the Quantitative Light-Induced Fluorescence (QLF) program. Specimens were divided into two groups and demineralized for 48 or 96 h and QLF images were again acquired. Fluorescence radiance (FR) of both sound (FR(s)) and demineralized (FR(d)) enamel was determined using the QLF program. Change in fluorescence radiance (Delta FR, %) was determined as follows: Delta FR = FR(d)/FR(s) x 100. One thin section was obtained for transverse microradiography analysis. Enamel and dentin thickness of the thin sections were measured. There was a good correlation between QLF parameters and enamel thickness. It can be concluded that the FR observed for similar lesions depends on the actual enamel thickness. PMID- 12566634 TI - Clinical study of the use of the laser fluorescence device DIAGNOdent for detection of occlusal caries in children. AB - Diagnosis of non-cavitated occlusal caries is generally considered problematic. The aim of this study was to evaluate the utility of the laser fluorescence device DIAGNOdent as part of routine dental check-ups in children. A total of 613 occlusal surfaces on permanent molars and 436 surfaces on primary molars in children aged 7-8 (n = 55) and 13-14 years (n = 54) were examined visually, using DIAGNOdent and for most 13- to 14-year-olds also using radiographs. The teeth were not cleaned professionally before examination. To provide a gold standard for carious teeth, the depth of the caries lesion was determined in those lesions that were judged visually to need opening by drilling (51 teeth). The higher the visual score, the higher were the mean DIAGNOdent values, but in each visual category the variation among values was large. The presence of a clear sealant did not affect DIAGNOdent measurements. The values for permanent and primary molars differed slightly. For permanent teeth, Youden's index value was highest (60%) at a cut-off value of 30, which indicates that the overall performance for DIAGNOdent in detecting dentinal caries was best at this point. Using this cut off, sensitivity was 92% and specificity 69% with visual examination for validation, while sensitivity was 92% and specificity 82% with validation by fissure opening. Of the three methods, radiographic examination was the least accurate. In routine dental check-ups of children, DIAGNOdent appears to be useful as an adjunct to visual examination. PMID- 12566636 TI - Radiographic dentinal caries and its progression in occlusal surfaces in Dutch 17 year-olds: a 6-year longitudinal study. AB - In this study, the status of 705 occlusal surfaces of first and second molars of 90 17-year-olds was analysed longitudinally in a 6-year follow-up, using a combination of clinical and radiographic information. Clinical data were derived from an epidemiological project. Each of two examiners judged all bitewing radiographs. The overall Cohen's kappa for interexaminer agreement of radiographic assessments was 0.77. Between the age of 17 and 23 years, about one third of sound occlusal surfaces developed a new dentinal radiolucency, and over 70% of existing radiolucencies showed progression, both irrespective of the presence of a sealant. In both examination years, almost 20% of the restored surfaces showed signs of a dentinal radiolucency. It was concluded that at the age of 17 occlusal surfaces are still highly susceptible to new dentine caries and further progression of dentinal radiolucencies already present. PMID- 12566637 TI - Demineralization of Er:YAG and Er,Cr:YSGG laser-prepared enamel cavities in vitro. AB - The aim of the present study was to establish whether cavity preparation by means of an erbium laser with efficient water cooling is capable of reducing the susceptibility of the prepared dental enamel to demineralization and thus of achieving a potential caries-protective effect in the region of cavity margins. To this end, cavities limited to the enamel were prepared in the crowns of 10 teeth each using an Er:YAG (lambda = 2,940 nm) and an Er,Cr:YSGG laser (lambda = 2,780 nm). A control cavity prepared conventionally with a diamond drill in the same occlusal zone was assigned to each of these cavities. The specimens were then subjected to a pH-cycling caries model. Analysis was performed by quantitatively measuring the demineralization front under a polarized-light microscope. The results of the study showed that enamel cavities prepared with the erbium lasers used display a statistically significant acceleration of demineralization compared to conventionally prepared cavities (p < 0.01). The Er:YAG laser cavities revealed demineralization to a depth of 133.9 (SD 25.7) microm, while the value observed with the Er,Cr:YSGG laser was 133.8 (SD 25.8) microm. The depth of demineralization in the control groups was only 77.4 (SD 13.8) microm and 79.3 (SD 37.6) microm. No difference could be found between the two lasers (p = 0.98). Based on these in vitro tests, it cannot be assumed that use of the erbium laser for cavity preparation offers any advantages in terms of resistance to secondary caries in clinical practice. PMID- 12566639 TI - Different analytical approaches in an experimental cohort study on preventive measures for caries in adolescents. A comparison between incidence density and increment analysis. AB - AIM: To use different analytical approaches to data from a longitudinal, quasi experimental study evaluating caries-preventive measures in order to estimate the differences in results due to the analytical tool used. METHODS: 1,165 patients at high risk for caries underwent yearly examinations in 1996, 1997 and 1998 and randomly received one out of four tested preventive measures. Two analytical methods were used: (1) increment was calculated using the indices DMFS(a) and D(e)MFS(a), (2) incidence density was calculated for each individual using new approximal dentin lesions or dentin and enamel lesions in the numerator and the risk time for the surfaces in the denumerator. The Poisson regression with overdispersion was used for multivariate modelling of the outcomes. RESULTS: In all analyses of approximal dentine as well as dentine and enamel caries bad oral health at baseline increased the risk of new approximal dentine caries. In the incidence analysis poor oral health behaviour also increased the risk. CONCLUSION: Since the difference between the two analytical methods are small for young individuals the method of preference would be increment analysis. PMID- 12566638 TI - Caries risk tests and salivary levels of immunoglobulins to Streptococcus mutans and Candida albicans in mouthbreathing syndrome patients. AB - The aim of this study was to compare microbiological and salivary variables possibly related to caries risk in treated and untreated mouthbreathing syndrome (MBS) children and control children. Thirty control children, 30 mouthbreathers and 25 treated mouthbreathers were studied for the numbers of lactobacilli, mutans streptococci and yeasts in their saliva. Snyder's test, salivary flow and buffering capacity were also evaluated. Levels of immunoglobulins to Candida albicans and Streptococcus mutans in the saliva were quantified using ELISA. Considering the results obtained for the microbiological and salivary caries risk tests, no significant differences were observed among the proportions of patients with small/negative and high/moderate caries risk in the studied groups. The level of IgG to S. mutans was significantly higher in the treated MBS group in relation to MBS patients. On the other hand, the median anti-S. mutans IgM level was lower in the treated MBS patients than in the other groups. For the studied anti-Candida immunoglobulins, IgM level was significantly lower in the treated MBS group than in the other groups. No differences were observed for anti-S. mutans and anti-Candida IgA levels among the groups. The findings suggest that mouthbreathing cannot be considered a risk factor for dental caries. PMID- 12566641 TI - Host-derived proteinases and degradation of dentine collagen in situ. AB - Dentine root caries is a process of demineralization and degradation of the organic matrix by proteinases. In this in situ study, the presence and activity of the matrix metalloproteinases 1, 2 and 9 (MMP-1, MMP-2, MMP-9) in saliva and in completely demineralized dentine specimens were investigated. Furthermore, the activity of cathepsin B was determined in saliva. A correlation between these enzymes and the level of degraded collagen was investigated. Demineralized dentine specimens were mounted in the partial prosthesis of 17 volunteers. Saliva samples were taken at 0, 2 and 4 weeks. After 4 weeks, the enzymes were extracted from the dentine specimens and the collagen loss was assessed. The collagen loss varied between 0 and 40.3%. Zymography of the saliva and the dentine extract samples showed that (pro-)MMP-2 and (pro-)MMP-9 were present. The levels of active MMPs were assessed, using fluorogenic MMP-specific substrates. All but 3 of the 51 saliva samples showed MMP-1 activity ranging from 1.5 to 101.1 relative fluorescence units (RFU)/s. Forty-eight saliva samples showed gelatinolytic MMP 2/MMP-9 activity (1.7-141.1 RFU/s). MMP-1 activity was shown in all dentine extracts varying between 3.5 and 295.0 RFU/s. From the dentine extracts, 15 showed MMP-2/MMP-9 activity (0.2-13.7 RFU/s). The MMP activity from both saliva and dentine extracts did not correlate with the collagen loss. The activity of salivary cathepsin B varied from 4.8 to 42.2 arbitrary units/min. A positive correlation was found between salivary MMP activity and cathepsin B activity. This study revealed that gelatinolytic enzyme activity was present both in saliva and dentine collagen. No correlation could be observed, however, between the level of enzyme activity and the collagen loss of the dentine specimens. PMID- 12566640 TI - Influence of short-term sucrose exposure on plaque acidogenicity and cariogenic microflora in individuals with different levels of mutans streptococci. AB - The aim was to study the short-term effect of a varying sucrose exposure on plaque acidogenicity and cariogenic microflora in two groups of subjects with different levels of mutans streptococci (MS). Eight subjects with low (<10(4)) and 8 with high (>10(6)) numbers of MS per millilitre saliva participated. Three 7-day test periods were conducted. During two of these, the subjects rinsed either 5 or 10 times daily with 10% sucrose; the third period without any mouth rinses served as control. The subjects refrained from oral hygiene during the last 3 days of each test period. On day 7, the following parameters were measured: plaque pH after a sucrose rinse, numbers of MS, Streptococcus sanguis and lactobacilli in saliva, percent MS in plaque and plaque index. The results revealed that plaque acidogenicity was more pronounced for the high-MS group compared to the low-MS group after all three test periods, i.e. lower resting pH, deeper pH falls and a lower final pH. For both groups, the greatest pH-lowering capacity of plaque was found after the period with 10 sucrose rinses/day. An increase in bacterial counts was noted for both groups during the test periods with the 5- and 10-time rinse regimen; this increase was larger for the high-MS group compared to the group with low MS counts. The highest plaque index was, irrespective of the test period, found for the high-MS group. PMID- 12566642 TI - Fluoride concentration in saliva after use of oral hygiene products. AB - The purpose of this in vivo, single-blind, randomized study was to compare fluoride concentrations in saliva of patients treated with oral hygiene products containing different fluoride salts. The study involved 104 students attending the University of Sassari. Participants were subdivided: group A used a sodium monofluorophosphate (NaMFP) toothpaste; groups B and C used an amine fluoride (AmF) toothpaste; group D used a toothpaste and a mouthwash both based on AmF, and group E used a toothpaste and a varnish both on an NaMFP base. Samples of unstimulated saliva were collected at baseline (t(0)), at the end of the 20 days' treatment phase (t(1)) and after 24 h, during which the volunteers refrained from any oral hygiene measure (t(2)). Saliva fluoride concentrations were measured using an ion-specific electrode. All measurements were made in triplicate and analysed statistically using ANOVA. In saliva, the mean fluoride concentration increased significantly in each treatment group. In conclusion, the fluoride concentration in saliva can be maintained to an optimal therapeutic level with the regular use of fluoridated products. PMID- 12566643 TI - Synergy of binary poly(oxypropylene-oxyethylene) copolymers in reducing retention of Streptococcus sanguis to hydroxyapatite. AB - Binary poly(oxypropylene-oxyethylene) block copolymer systems have been investigated as a non-bactericidal approach to reduce the retention of an oral bacterium, Streptococcus sanguis. Using a previously validated hydroxyapatite coated microtitre model to simulate the tooth, the performance of copolymer pairs was measured experimentally. A synergy index and an efficacy index were defined to describe the reduction of bacterial retention by the binary systems that comprise the copolymer pairs. Relationships between the synergy and efficacy indices and their associated compositions are given. The results obtained have been rationalised using a previously developed theoretical approach in conjunction with the binding energetics of species to surfaces. PMID- 12566644 TI - Reverse remodeling of the failing human heart with mechanical unloading. Emerging concepts and unanswered questions. AB - Anecdotal and clinical evidence suggests that mechanical unloading may restore cardiac function by inducing changes in the biological properties of the failing heart. This challenges the notion that the progression of end-stage heart failure is an irreversible process ending in either death of the patient or transplantation. Although it is still not clear how mechanical unloading of the failing heart improves cardiac function, the process likely involves adaptive responses of cardiac myocytes on the cellular, extracellular, and subcellular levels. PMID- 12566645 TI - Natural history and predictors of temporary epicardial pacemaker wire function in patients after open heart surgery. AB - STUDY OBJECTIVES: To assess the short-term reliability of temporary epicardial pacing wires in patients after open heart surgery and to determine the influence of preoperative factors on the performance of these wires. PATIENTS AND METHODS: In this prospective, longitudinal cohort study, 60 patients (41 men) with a mean age of 66.8 +/- 8.9 years were studied for 5.3 +/- 1.1 days postoperatively. Each patient's history, physical findings, hemodynamics and echocardiographic data were analyzed. Postoperatively, capture threshold, the electrogram amplitude and slew rate, and the lead impedance were determined daily in both chambers. RESULTS: Both atrial and ventricular thresholds increased significantly (p < 0.001) by the 4th postoperative day. The P-wave amplitudes were consistently less than half of the R-wave amplitudes. Both P- and R-wave amplitudes deteriorated significantly by the 2nd postoperative day (p < 0.001 and p < 0.02, respectively). Atrial and ventricular slew rates and lead impedance decreased significantly during the observation period (p < 0.002). CONCLUSION: Although both atrial and ventricular temporary epicardial leads are reliable for short term use, their function deteriorates on a daily basis. Perioperative factors are generally not helpful in predicting the performance of temporary epicardial pacing wires. PMID- 12566646 TI - Coronary artery bypass surgery in heart failure patients with chronic reversible and irreversible myocardial dysfunction: effect on heart rate variability. AB - It is unknown whether surgical revascularization of viable and chronic reversibly dysfunctional myocardium in patients with heart failure is associated with a favorable effect on the autonomic control of heart rate. We studied 38 coronary artery bypass surgery (CABG) patients with an ejection fraction of 36 +/- 7%. Before CABG, the patients underwent 48-hour ambulatory electrocardiographic monitoring to measure heart rate variability (HRV), echocardiography to assess left ventricular function, and positron emission tomography and low-dose dobutamine echocardiography to assess viability. Six months after CABG, ambulatory electrocardiographic monitoring and echocardiography were repeated to assess HRV and recovery of left ventricular function. In spite of viable myocardium and recovery of left ventricular function following CABG, HRV indices were reduced 6 months postoperatively. A potential prognostic benefit gained by revascularizing patients with viability may occur in spite of deteriorated autonomic control of heart rate. PMID- 12566647 TI - Comparison between percutaneous balloon valvuloplasty and open commissurotomy for mitral stenosis. A prospective and randomized study. AB - Eighty patients with tight and pliable mitral stenosis were randomized in a prospective trial comparing percutaneous balloon valvuloplasty and open commissurotomy. Mitral valve area increased significantly in both groups immediately after the procedures. However, a decrease in mitral valve area occurred in both groups at 6, 12 and 24 months during the follow-up. There was no death in either group. Two patients had significant mitral regurgitation after percutaneous balloon valvuloplasty. After 24 months, all patients in the commissurotomy group and 97.4% of the patients in the balloon valvuloplasty group were in New York Heart Association functional class I or II. PMID- 12566648 TI - Yield of 5,536 diagnostic coronary arteriographies: results from a data registry. AB - The number of coronary arteriographies (CAG) has increased tremendously all over the industrialised world over the past years. Even though the potential benefit is high in patients with angina pectoris, for example, with expected life prolongation in case of three-vessel disease or left main stenosis with subsequent coronary artery bypass grafting (CABG), the indication for treatment is not as dramatic in all patients and at the same time CAG is a procedure with at least, some risk, and costs are not negligible. It is therefore pertinent, and hence also the purpose of the present analysis, to make observations on indications, clinical and angiographic findings and their combinations which could be helpful in clinical practice/decision-making. Furthermore, the purpose was to analyse the consequences of CAG with respect to revascularisations such as percutaneous transluminal coronary angioplasty (PTCA) or CABG. PMID- 12566649 TI - Characteristics, treatment and outcome of patients with non-ST-elevation acute coronary syndromes and multivessel coronary artery disease: observations from PURSUIT (platelet glycoprotein IIb/IIIa in unstable angina: receptor suppression using integrelin therapy). AB - BACKGROUND: The 6-month clinical outcome of patients with multivessel disease enrolled in PURSUIT (Platelet Glycoprotein IIb/IIIa in Unstable Angina: Receptor Suppression Using Integrilin Therapy) is described. Patients with complete angiography data were included; multivessel disease was stratified according to the treatment strategy applied early during hospitalization, i.e. medical treatment, percutaneous coronary intervention (PCI) (balloon), PCI (stent), or coronary artery bypass grafting (CABG). METHODS: Patients were divided into three groups according to the treatment strategy applied during the first 30 days of enrolment. Patients who did not undergo a percutaneous or surgical coronary intervention were classified as medically treated. Patients who underwent a PCI (prior to a possible CABG) were separated from those who underwent a CABG (prior to a possible PCI). The PCI group was further subdivided: patients receiving >/=1 coronary stents were separated from those in whom no stents were used. RESULTS: The mortality rate at 30 days was 6.7, 3.9, 2.4 and 4.8% for the medical treatment, PCI (balloon), PCI (stent) and CABG groups, respectively (p value = 0.002). Differences as observed at 30 days were still present at 6-month follow up with 11.1, 5.8, 5.5 and 6.5% mortality event rates for the aforementioned groups (p value = 0.002). The 30-day myocardial infarction (MI) rate according to the opinion of the Clinical Events Committee was lower among medically than non medically treated patients, with the highest event rate observed in the CABG group (27.7%). Approximately half of the MIs in the PCI and CABG subgroups occurred within 48 h after the procedure. CONCLUSIONS: The observed differences in clinical outcomes are explained by an imbalance in baseline characteristics and comorbid conditions between the analyzed groups of patients. PMID- 12566650 TI - Prediction of left ventricular functional recovery by dobutamine echocardiography, F-18 deoxyglucose or 99mTc sestamibi nuclear imaging in patients with chronic myocardial infarction. AB - BACKGROUND: Currently, several modalities are available to predict viability, however, studies comparing various modalities validated by functional recovery after revascularization are scarce. This study analyzed the relative merits of low-dose dobutamine echocardiography, F-18 deoxyglucose (FDG) positron emission tomography (PET) and (99m)Tc sestamibi single-photon emission computed tomography to predict functional recovery after revascularization in patients with chronic myocardial infarction. METHODS: Patients with chronic coronary occlusion (duration: 3.1 +/- 4.8 years) and impaired left ventricular function (ejection fraction: 42 +/- 13%) underwent low-dose dobutamine echocardiography (20 microg/kg/min), FDG-PET and (99m)Tc sestamibi imaging before revascularization. Revascularization was performed irrespective of any viability data. Follow-up angiography was obtained 4.8 +/- 2.5 months after revascularization. RESULTS: Viability analysis was performed in 34 patients with patent target vessel at follow-up, of whom 9 (27%) exhibited functional recovery on left ventricular angiography. For dobutamine echocardiography, improvement of >/=2 adjacent akinetic segments resulted in improved sensitivity of 89% and specificity of 80% to predict functional recovery. For glucose metabolism, FDG uptake >55% was an optimal threshold yielding a sensitivity of 89% and a specificity of 68%. With respect to perfusion, (99m)Tc sestamibi uptake >60% was the best cutoff resulting in a sensitivity and a specificity of 56 and 88%, respectively. A concordant match of FDG >55% and of (99m)Tc sestamibi >50% resulted in optimized sensitivity (78%) and specificity (80%) with dual imaging. CONCLUSIONS: Recovery of chronically dysfunctional myocardium can be predicted with high accuracy by stimulation of contractile reserve or by concordant match of preserved glucose metabolism and residual perfusion. PMID- 12566651 TI - The molecular cardiology laboratory at baylor college of medicine. PMID- 12566652 TI - Dietary measures in individuals of South asian ancestry with cardiovascular disorders. PMID- 12566653 TI - Severe mitral valve regurgitation associated with dexfenfluramine use. PMID- 12566654 TI - Congestive heart failure caused by digitalis toxicity in an elderly man taking a licorice-containing chinese herbal laxative. PMID- 12566655 TI - Blood purification in non-renal critical illness. AB - Extracorporeal therapies designed to remove substances from the circulation now include hemodialysis, hemofiltration, hemoadsorption, plasma filtration, cell based therapies and combinations of any of the above. In recent years, there have been considerable advances in our understanding and technical capabilities, but consensus over the optimal way, and under what conditions to use these therapies does not exist. Consequently, we have initiated a series of conferences under the auspices of the Acute Dialysis Quality Initiative (ADQI). ADQI aims at establishing an evidence-based appraisal and set of consensus recommendations to standardize care and direct further research. The results of previous consensus conferences are available on-line at www.ADQI.net. ADQI conferences have focused on research and management of renal disease. However, extracorporeal therapies are being used or investigated in the management of many other disease processes including systemic inflammation, liver disease, cardiac disease and thrombotic diseases. PMID- 12566656 TI - Recent concepts in the molecular biology of the peritoneal membrane - implications for more biocompatible dialysis solutions. AB - This paper reviews some important recent findings on the molecular biology of the peritoneal membrane. It attempts to correlate in vitro and in vivo experimental results with the possible clinical consequences. The most common functional alteration during long-term CAPD is increased peritoneal small-solute transport rate, resulting in impaired ultrafiltration and decreased dialysis efficiency. This contribution first discusses the most relevant advances in the biochemistry and molecular biology of the peritoneal membrane following peritonitis and as consequence of the continuous exposure to unphysiological dialysis fluids. In a second part the preliminary experimental and clinical experience with more biocompatible fluids is summarized. The most relevant structural and functional alterations of the membrane following repeated peritonitis is the consequence of the response of the peritoneum to infective organisms involving the inflammatory cytokines and the interaction between membrane resident cell populations: macrophages, mesothelial cells and fibroblasts. In this setting, human biopsy studies and animal experiments have identified an increase in the peritoneal associated vasculature, which seems to be the primary cause of increased solute transport. The structural and functional alterations in the membrane in long-term peritoneal dialysis are thought to be the consequence of the toxicity of glucose, either directly or indirectly through the generation of glucose degradation products or the formation of advanced glycation end-products. In particular, an important role for vascular endothelial growth factor and nitric oxide as downstream mediators of the alterations has been suggested. Finally, the last part of this paper reviews the actual and future research aimed at an amelioration of the biocompatibility of the dialysis fluids. Replacing glucose by other osmotic agents, changing the sterilization process, replacing the lactate buffer by bicarbonate, blocking the formation of reactive carbonyl products and of the neoangiogenesis are the most promising changes to enhance the biocompatibility. Finally, gene therapy may in the future have an important contribution. Ex vivo gene therapy involves harvesting peritoneum samples to isolate mesothelial cells that will be genetically modified before re implantation into the peritoneal cavity. PMID- 12566657 TI - PD and the future: the role of PD in the overall management of ESRD. AB - Given the epidemic growth of ESRD and the high costs of dialysis therapy, there is an urgent need to define the optimal dialysis modality distribution. The factors influencing dialysis modality distribution are complex and not well understood. Based on survey results that have assessed the attitudes of nephrologists in Canada, the USA and the UK, it appears that there is underutilization of both peritoneal dialysis (PD) and home hemodialysis (HD) in these countries. Nephrologists have a positive attitude towards home therapies, and do not appear to be biased against PD. A planned approach to dialysis initiation, with active promotion of PD and home HD as the initial dialysis modality for suitable patients, would be expected to revive usage of home PD and allow for the emergence of daily home HD as a significant and exciting new modality. PMID- 12566658 TI - Inflammation and atherosclerosis in end-stage renal disease. AB - Atherosclerosis is a multifaceted process which may be initiated by various insults to vascular endothelium. Independently of the nature of the offending factor, the endothelial dysfunction that results from the initial insult is characterized by increased adhesiveness of the endothelium to leukocytes and platelets and by the synthesis of vasoactive molecules, cytokines and procoagulant factors. This defensive response is characterized by classical inflammatory changes and may lead to plaque formation, luminal obstruction and plaque rupture. Factors involved in arterial damage in end-stage renal disease (ESRD) span from classical risk factors to disease-peculiar factors (anemia, secondary hyperparathyroidism and exposure to bioincompatible dialysis membranes and/or contaminated dialysis fluid) and to emerging and novel risk factors such as hyperhomocysteinemia, infections and accumulation of the endogenous inhibitor of NO synthase, asymmetric dimethylarginine (ADMA). There is strong and consistent evidence that acute phase reactants like C-reactive protein and cytokines like IL-beta, TNF-alpha and IL-6 are independently associated with death and atherosclerosis in ESRD patients. The experimental and epidemiological data collected thus far coherently show that endothelial dysfunction resulting from inflammation may promote abnormal vascular behavior and thrombosis in ESRD. There are several possible therapeutic approaches for reducing the risk excess associated with inflammation in ESRD. These possibilities range from drugs interfering with the angiotensin system or with adrenergic activity to anti inflammatory and antilipid agents to vitamins, antioxidants, to the amino acid precursor of nitric oxide, L-arginine, and perhaps to antibiotics. The intellectual framework is well delineated but very few controlled trials have been performed or are underway in patients with ESRD. PMID- 12566659 TI - The ADEMEX study and PD adequacy. AB - The ADEMEX study was a prospective, randomized, controlled, interventional trial that evaluated the effect of an increase in peritoneal clearance on the relative risk of death for patients on CAPD. The results suggest that over the range of solute clearance studied, increasing peritoneal solute clearance had no beneficial effect on survival. Residual renal function did predict outcome. The findings are clinically relevant, but there are some limitations of the study that may limit the generalizability of the results. First of all, exclusion criteria were likely to result in the exclusion of rapid transporters and small patients, the subgroup of patients found to have an increased relative risk of death on PD in other studies. Secondly, ADEMEX evaluated the effect of an increase in small solute, not middle molecular weight solute clearances on outcome. Lastly, ADEMEX did not evaluate outcomes for patients on cycler therapies, the most popular form of PD in use currently. The data confirms (a) that one size prescription does not fit all; (b) that many patients below current NKF-DOQI targets for small solute clearance may be adequately dialyzed, and (c) it provides us with evidence-based data that national societies can consider using when preparing for the next revisions of their guidelines. PMID- 12566660 TI - ADEMEX and HEMO trials: where are we going? AB - Doubt has remained as to whether or not the K/DOQI recommended targets for adequacy of dialysis for peritoneal dialysis patients is appropriate (weekly Kt/V 2 + creatinine clearance 50-60 l/1.73 m(2)). The ADEMEX trial can be interpreted as indicating that lower targets might be acceptable. The HEMO trial, not yet published but presented in April 2002, casts doubts on the advantages of achieving higher than recommended small solute clearance targets. Taken together, these trials require that we broaden our concept of adequacy. There is also a risk of complacency with respect to dialysis adequacy because of these trials and this would be unwise. PMID- 12566661 TI - Current studies on oxidant stress in dialysis. AB - Breath ethane measurements in hemodialysis indicate that a portion of these patients suffer increased oxidant stress, consistent with findings using other methods for oxidant stress determination. Loosely-bound iron definitely appears in the bloodstream when substantial doses of IV iron are administered, since transferrin is fully saturated, but our investigations generally do not show short-term oxidant stress from this treatment. If small doses of IV iron are utilized, transferrin saturation can be avoided, and risk is minimized. The vitamin C status of hemodialysis patients is usually lower than the general population, and the impact of this deficiency must be assessed in controlled investigations. Various interventions, including the vitamin E-bonded dialyzer and dietary antioxidant supplements, may ameliorate a portion of the oxidant stress in hemodialysis patients. PMID- 12566662 TI - A kinetic model of inorganic phosphorus mass balance in hemodialysis therapy. AB - BACKGROUND: There is growing evidence that inorganic phosphorus (iP) accumulation in tissues (dTiP/dt) is a risk factor for cardiac death in hemodialysis therapy (HD). The factors controlling iP mass balance in HD are dietary intake (GiP), removal by binders (JbiP) and removal by dialysis (JdiP). If iP accumulation is to be minimized, it will be necessary to regularly monitor and optimize GiP, JbiP and JdiP in individual patients. We have developed a kinetic model (iPKM) designed to monitor these three parameters of iP mass balance in individual patients and report here preliminary evaluation of the model in 23 HD patients. METHODS: GiP was calculated from PCR measured with urea kinetics; JdiP was calculated from the product of dialyzer plasma water clearance (K(pwiP)) and time average plasma iP concentration (TACiP) and treatment time (t); a new iP concentration parameter (nTAC(iP), the TACiP normalized to predialysis CoiP) was devised and shown to be a highly predictable function of the form nTAC(iP) = 1 - alpha(1 - exp[-betaK(pwiP). t/ViP]), where the coefficients alpha and beta are calculated for each patient from 2 measure values for nTAC(iP), K(pwiP).t/ViP early and late in dialysis; we measured 8-10 serial values for nTAC(iP), K(pwiP). t/ViP over a single dialysis in 23 patients; the expression derived for iP mass balance is DeltaTiP = 12(PCR) - [K(pwiP)(t) (N/7)][CoiP(1 - alpha(1 - exp[ beta(Kt/ViP)]))] - k(b).Nb. RESULTS: Calculated nTAC(iP) = 1.01(measured nTAC(iP)), r = 0.98, n = 213; calculated JdiP = 0.66(measured total dialysate iP) + 358, n = 23, r = 0.88, p < 0.001. Evaluation of 10 daily HD patients (DD) and 13 3 times weekly patients with the model predicted the number of binders required very well and showed that the much higher binder requirement observed in these DD patients was due to much higher NPCR (1.3 vs. 0.96). CONCLUSION: These results are very encouraging that it may be possible to monitor the individual effects of diet, dialysis and binders in HD and thus optimize these parameters of iP mass balance and reduce phosphate accumulation in tissues. PMID- 12566663 TI - Affinity hemodialysis for antiviral therapy. II. Removal of HIV-1 viral proteins from cell culture supernatants and whole blood. AB - BACKGROUND: HIV-1 gp120 may play a role in the progression from HIV infection to AIDS. AIMS: We investigated affinity hemodialysis for removing gp120 from cell culture and whole blood. METHODS: Anti-gp120 antibodies covalently coupled to agarose beads were packed into columns or hollow-fiber hemodialysis cartridges. Supernatants from HIV-infected HL2/3 cells or gp120 containing whole blood were pumped over the columns and gp120 measured by ELISA. RESULTS: Anti-gp120 agarose removed approximately 90% of HIV-1 gp120 from HL2/3 cultures in 30-60 min. Capture was antibody-dependent (F105 > IDX 1121 > ABI 13-108). Affinity hemodialysis also efficiently captured gp120 from buffer in a first-order, flow rate-dependent fashion (t((1/2)) = 13 min at 0.9 ml/min). Clearance was faster than calculated diffusion (t((1/2)) approximately 2.5 h) suggesting significant convective transport. gp120 removal from blood was slower (t((1/2)) = 1.4 h). CONCLUSION: Affinity hemodialysis efficiently clears gp120 from cell culture fluids and blood and may be useful in slowing the progression to AIDS. PMID- 12566664 TI - Renal cell therapy is associated with dynamic and individualized responses in patients with acute renal failure. AB - BACKGROUND: Renal cell therapy in conjunction with continuous hemofiltration techniques may provide important cellular metabolic activities to patients with acute renal failure (ARF) and may thereby change the natural history of this disorder. The development of a tissue-engineered bioartificial kidney consisting of a conventional hemofiltration cartridge in series with a renal tubule assist device (RAD) containing 10(9) human renal proximal tubule cells provides an opportunity to evaluate this form of therapy in patients with ARF in the intensive care unit. METHODS: Nine patients with ARF and multi-organ systems failure (MOSF) have been treated so far with a tissue-engineered kidney in an FDA approved Phase I/II clinical study currently underway. Acute physiologic parameters and serum cytokine levels were assessed before, during and after treatment with a bioartificial kidney. RESULTS: Use of the RAD in this clinical setting demonstrates maintenance of cell viability and functionality. Cardiovascular stability appears to be maintained during RAD treatment. Human tubule cells in the RAD demonstrated differentiated metabolic and endocrinologic activity. Acute physiologic and plasma cytokine data demonstrate that renal cell therapy is associated with rapid and variable responses in patients with ARF and MOSF. CONCLUSION: The initial clinical experience with the bioartificial kidney and the RAD suggests that renal tubule cell therapy may provide a dynamic and individualized treatment program as assessed by acute physiologic and biochemical indices. PMID- 12566665 TI - Slow continuous intracorporeal plasmapheresis for acute fluid overload. AB - Intermittent dialysis is still the predominant treatment for acute or chronic renal insufficiency in the USA despite increasing evidence that slower and longer fluid management therapies are more beneficial to the patient. We have investigated the use of slow continuous intracorporeal plasmapheresis (SCIP) as a more efficient and hemodynamically stable alternative means of treating acute fluid overload. In this paper we discuss preliminary observations on the safety of SCIP catheter insertion, fluid removal, extraction and pathology in Yorkshire pigs. SCIP catheters removed plasma for extracorporeal plasma water removal without significant gross or histopathological changes. Blood chemistry and cell counts remained stable during therapy. Toxicological studies indicated no pyrogenicity, hemolysis, cytotoxicity, acute systemic toxicity, delayed-type hypersensitivity, or blood recalcification coagulation inhibition. Intracutaneous extracts caused only mild irritation. SCIP therapy appears to be safe for use in the removal of plasma and plasma water from experimental animals. PMID- 12566666 TI - Sorbents in acute renal failure and the systemic inflammatory response syndrome. AB - Renal replacement therapy in acute renal failure is currently focused on the use of modifications of dialysis (continuous arteriovenous hemofiltration and hemodiafiltration) to remove middle molecular weight toxins, consisting of small proteins, and cytokines involved in the systemic inflammatory response syndrome (SIRS). Conventional high-flux dialyzers are not efficient at removing these molecules, prompting the investigation of sorbents to augment or replace dialysis. Sorbents have been developed to modulate SIRS by targeting cytokines such as IL-1, IL-6, IL-10, IL-18 and TNF, among others. Extensive pre-clinical studies are underway to demonstrate the clinical utility and safety of either adding sorbent hemoadsorption devices to hemodialysis, or the use of such devices alone in SIRS, sepsis, acute renal failure, cardiopulmonary bypass and end-stage renal disease. PMID- 12566667 TI - Long nocturnal dialysis. AB - BACKGROUND/AIMS: The continuous growth of the dialysis pool in our unit induced us to organize a third long nocturnal dialysis (LND) session, considering the excellent survival and rehabilitation results reported with this method. This paper analyzes the results and assesses the role of LND among the different dialytic treatment options. METHODS: Out of 18 patients on LND, 13 (12 males and 1 female, mean age 52 +/- 13 years, time on dialysis 21.8 +/- 23.8 months) with >6 months' experience were studied, and 9 underwent a further metabolic evaluation. LND was performed using 1- to 1.4-m(2) Hemophan membranes, bicarbonate buffer, 200-250 ml/min blood flow, and 300-500 ml/min dialysate flow, 8 h three times a week. Kt/V and protein catabolic rate (3-point classic urea kinetics), postdialytic weight, serum albumin, total protein, hemoglobin, Ca(2+), phosphate, intact parathyroid hormone, bioimpedance body water, blood pressure, and drug use (antihypertensives, phosphate binders, erythropoietin, vitamin D, hypnotics) were evaluated in each patient during hemodialysis and LND. In the metabolic study (done twice), sodium (compared with the Kimura model), potassium, phosphate, and urea were analyzed in blood and inlet and outlet dialysate after 0, 2, 4, 6, and 8 h. RESULTS: The mortality was low (1 death every 247 patient months). After 19 +/- 8.1 months of LND, the postdialytic weight rose from 68.5 +/- 9.6 to 70.8 +/- 10.7 kg (p < or = 0.01), and the hemoglobin concentration rose from 10.8 +/- 2.2 to 11.8 +/- 1.8 g/dl (p < or = 0.05); phosphate dropped from 5.6 +/- 2.0 to 4.4 +/- 1.3 mg/ dl (p < or = 0.01) and the systolic blood pressure from 152 +/- 15 to 143 +/- 19 mm Hg (p < or = 0.05). In the metabolic study, the sodium profile was significantly lower during the last 4 h than in the Kimura model. The potassium concentration, stable between 4 and 6 h, rose against the gradient during the last 2-hour period. The behavior of sodium and potassium during the last part of the dialysis session can be taken to indicate exhaustion of the sodium/potassium pump. Phosphate showed a gradual reduction with no intradialytic and only a moderate postdialytic rebound. The postdialytic urea rebound was 23.4%. CONCLUSIONS: LND is a useful additional tool for nephrologists in treating chronic renal failure, it is easy to organize, and it shows overall good results. Together with other dialysis methods, this schedule permits individualized treatment for each uremic patient. PMID- 12566668 TI - Advanced glycation end products and advanced oxidation protein products in hemodialyzed patients. AB - Chronic renal failure is associated with increased oxidative and carbonyl stresses that contribute to long-term uremic complications. In our study, we determined two markers of these stresses--AGEs (advanced glycation end products) and AOPP (advanced oxidation protein products)--in chronic hemodialysis patients in order to find out their relationship to the dialysis treatment. Plasmas of 20 hemodialyzed patients treated with modified cellulose membranes were examined at 0 and 15 min and at the end (i.e. after 4 h) of the dialysis session. AGEs were estimated using a spectrofluorometric method (excitation 350 nm, emission 440 nm) and are expressed in AU (arbitrary units)/g protein. AOPP were determined spectrophotometrically (absorbance at 340 nm) and are expressed in chloramine units per gram of protein (micromol/g). AOPP decrease slightly from 0 to 15 min of the dialysis procedure (4.0 +/- 1.5 vs. 3.0 +/- 0.9 micromol/g, p < 0.01). However, they are increased at the end of the session (5.0 +/- 2.1 micromol/l vs. 15 min, p < 0.01, not significant vs. beginning). On the other hand, AGEs decrease continuously from the beginning to the end of the session (mainly in the first minutes of the dialysis) (1.52 +/- 0.34 x 10(4) AU/g at 0 min, 1.39 +/- 0.33 x 10(4) AU/g at 15 min, p < 0.001 vs. beginning, 1.30 +/- 0.33 x 10(4) AU/g at the end, p < 0.001 vs. beginning, not significant vs. 15 min). Neither AGEs nor AOPP correlate with the age of hemodialyzed patients and with the number of years of the dialysis treatment. AOPP correlate with AGEs before the dialysis session (r = 0.62, p < 0.05) but not after the session (r = 0.29, not significant). According to our results, AGEs may serve more as a marker of chronic damage while AOPP may better describe acute oxidative stress during the dialysis treatment. PMID- 12566669 TI - 8-iso-prostaglandin F2alpha as a useful clinical biomarker of oxidative stress in ESRD patients. AB - BACKGROUND/AIMS: Chronic renal failure is associated with elevated indices of oxidative stress. We tested the hypothesis that the in vivo formation of the F(2) isoprostane (8-iso-prostaglandin PGF(2alpha)), a bioactive product of arachidonic acid peroxidation, is enhanced in end-stage renal disease (ESRD) patients receiving hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD). METHODS: Plasma samples were obtained from 35 HD patients, 30 CAPD patients and 30 age- and sex-matched healthy subjects for measurement of immunoreactive 8-iso PGF(2alpha). RESULTS: Plasma 8-iso-PGF(2alpha) levels were significantly higher (p < 0.001) in HD and CAPD patients (346.3 +/- 132.4 pg/ml; range 49.8-870) than in age-matched control subjects (150.9 +/- 61.6 pg/ml; range 33.5-235). In addition, we also found that 8-iso-PGF(2alpha) concentration was significantly (p = 0.007) higher in HD patients (389.8 +/- 148.3 pg/ml) than in CAPD patients (254.3 +/- 76.6 pg/ml). Plasma 8-iso-PGF(2alpha) concentration was linearly correlated with serum haptoglobin, C-reactive protein (CRP) and plasma MDA (r = 0.58, p = 0.003; r = 0.29, p < 0.05 and r = 0.38, p < 0.05 respectively). On the other hand, plasma 8-iso-PGF(2alpha) levels were inversely associated with serum albumin and total cholesterol (r = -0.31 and r = -0.28, respectively; p < 0.05). CONCLUSIONS: We conclude that ESRD on both HD and CAPD is associated with increased formation of F(2)-isoprostanes, a correlate of enhanced lipid peroxidation. We also found that plasma 8-iso-PGF(2alpha) was casually related to some acute phase reactant proteins such as serum CRP, albumin and haptoglobin. This may provide an important biochemical link between lipid peroxidation, inflammation and accelerated atherosclerosis in the uremic milieu. PMID- 12566670 TI - Defective interleukin-10 synthesis by peripheral blood mononuclear cells among hemodialysis patients. AB - BACKGROUND: Interleukin-10 (IL-10), a potent regulatory monokine produced by activated mononuclear cells, provides an efficient autocrine mechanism for controlling proinflammatory cytokine synthesis. We hypothesized that defective synthesis of IL-10 could contribute to the inflammatory state in hemodialysis (HD) patients due to impaired feedback inhibition of proinflammatory cytokine production. METHODS: We compared peripheral blood mononuclear cell (PBMC) synthesis and transcription of IL-10 and TNF-alpha in 12 patients with end-stage renal disease on long-term maintenance HD and a control group of 10 healthy subjects. RESULTS: The synthesis of IL-10 by unstimulated PBMC was detectable in 5 of 12 (42%) HD patients as compared to 7 of 10 (70%) controls (p = 0.02). IL-10 synthesis in response to endotoxin (ET) by PBMC from HD patients was significantly lower when compared to the robust response in the control group (p = 0.008). Among the HD patients, there was a positive correlation between ET stimulated IL-10 synthesis and the duration of time on dialysis. Unstimulated and ET-stimulated synthesis of TNF-alpha by PBMC did not differ between the 2 groups. In the HD patients, there was an inverse correlation between TNF-alpha and IL-10 synthesis by ET-stimulated PBMC, suggesting a regulatory effect of IL-10 on PBMC TNF-alpha synthesis. There was also an inverse correlation between plasma albumin and ET-stimulated TNF-alpha synthesis by PBMC among HD patients. TNF-alpha mRNA expression did not differ in HD patients relative to healthy controls. In contrast, when IL-10 mRNA from ET-stimulated PBMC was quantified, there was marked difference between the 2 groups indicating a transcriptional defect in IL 10 synthesis in PBMC from HD patients. CONCLUSION: Our observations indicate a marked abnormality in IL-10 synthesis by PBMC from HD patients probably related to a transcriptional defect. Low PBMC IL-10 synthesis may contribute to a chronic inflammatory state in these patients by defective feedback inhibition of proinflammatory cytokine production. PMID- 12566671 TI - Correction of acidosis by hemodialysis: proposal of a correlation with urea kinetics. AB - BACKGROUND/AIMS: We examined the effect of hemodialysis (HD) on acid-base status and its relation to urea kinetics in clinically stable renal HD patients. The purpose of this study was to design a practical approach to monitoring the correction of acidosis, as it can be assisted by routine parameters of adequacy. METHODS: Blood samples were drawn immediately before and after HD from 46 chronic renal patients to determine electrolytes, blood gases, serum albumin and blood urea nitrogen (BUN). Additional measurements of pH and serum bicarbonate were done in 35 patients in the periods before, immediately after and 4 h after HD. The normalized protein catabolic rate (nPCR) was calculated and correlated with the serum albumin and bicarbonate values before HD. Equilibrated KT/V (eKT/V) and urea reduction ratio (URR) were calculated and correlated with the degree of bicarbonate correction, defined as DeltaHCO(3)(-). RESULTS: There was no correlation between nPCR and pre-HD HCO(3)(-), while there was a significant correlation between URR and eKT/V and DeltaHCO(3)(-) (p < 0.003). CONCLUSIONS: The Deltabicarbonate was well correlated with URR and eKT/V, and the study suggests that in standard HD the correction of acidosis may be related to target URR and eKT/V levels. PMID- 12566672 TI - Hemodynamics of venous cuff interposition in prosthetic arteriovenous fistulas for hemodialysis. AB - PURPOSE: The durability of prosthetic arteriovenous fistulas (AVF) for hemodialysis is jeopardized by thrombotic occlusions due to intimal hyperplastic stenoses. In arterial reconstructive surgery, peripheral arterial bypasses with prosthetic material benefit from a venous cuff at the distal anastomosis. Therefore, a study was performed to assess the effect of a venous cuff at the venous anastomosis of PTFE graft AVFs in terms of stenosis development, hemodynamics and patency rates. METHODS: A subset of 40 patients from a multicenter study were enrolled into the study, of which 20 patients were randomized for venous cuff interposition. Duplex measurements to detect stenoses and volume flows were performed at 6, 12, 26 and 52 weeks postoperatively. Relative distension (RD) and wall shear rate (WSR) were calculated by means of vessel wall Doppler tracking (VWDT). RESULTS: The total number of stenoses was significantly less in the cuff group (21 vs. 33; p = 0.045). This feature was found at the site of the venous anastomosis (cuff 5; no cuff 12). Volume flow, graft and efferent vein diameters, RD and WSR in the graft and efferent vein were comparable for both groups. WSR in the venous anastomosis tended to be lower in the cuff group (768 vs. 1,448 s(-1), p = 0.068). Volume flows and WSR were significantly lower in failing grafts. Patency rates were similar in both groups (primary patency 13 vs. 29%; secondary patency 78 vs. 67%). CONCLUSIONS: A venous cuff at the venous anastomosis of PTFE graft AVFs results in less stenoses, but improved patency rates could not be demonstrated. PMID- 12566673 TI - Vascular access for dialysis in the elderly. AB - AIMS: The aim of this study was to analyze early and late outcome of the first vascular accesses performed in the HGUGM of Madrid between 1992 and 1997, comparing the results in patients older and younger than 65 years. METHODS: Retrospective study. All the vascular accesses performed in patients without previous permanent angioaccess between January 1992 and March 1997 were studied. Early failure, complication rate (patient-year of follow-up), and cumulative patency rates were analyzed. RESULTS: The difference between grafts required in young (25%) and old patients (34%) was significant (p < 0.01). Complication rate in autologous fistulas was 0.07 per patient-year in the younger group and 0.12 per patient-year in the older (p < 0.01). Complication rate in grafts was 0.5 per patient-year in the younger group and 0.8 per patient-year in the older group (p < 0.05). There were no significant differences in patency rates for both autologous and graft accesses between patients younger or older than 65 years. Better primary patency (log-rank comparison between curves, p < 0.001), and lower complication rates (0.09 patient-year for autologous and 0.58 for grafts, p < 0.001), were observed in autologous fistulas in both groups. CONCLUSIONS: Autologous access is the best angioaccess for dialysis also in older patients and can be performed in most patients without a previous permanent access. Yet, we observed in our experience that more grafts were needed in patients older than 65 years. PMID- 12566674 TI - Estimation of filtration coefficients and circulating plasma volume by continuously monitoring hematocrit during hemodialysis. AB - BACKGROUND/AIMS: Filtration coefficients (Lp) and plasma volume were estimated in order to investigate whether suppressed Lp associates with intradialytic hypotension and/or diabetic nephropathy. METHODS: Twenty-one patients were evaluated. Nine patients were diabetic (DM) and 12 were nondiabetic (non-DM). Three of DM and 4 of non-DM were prone to dialysis-induced hypotension (hypo(+)) and others (hypo(-)) were not. Changes in hematocrit (Ht) were measured for 60 min after the start of ultrafiltration. Lp and plasma volume at the start of ultrafiltration (Vp0) were estimated to fit calculating values of Ht based on Schneditz's open two compartment model to actual value. RESULTS: There was no significant difference in the mean values of Lp/Vp0 either between hypo(+) and hypo(-) (0.87 +/- 0.37 vs. 1.24 +/- 0.48 ml/mm Hg.min.liter; n.s.) or between DM and non-DM (1.04 +/- 0.32 vs. 1.17 +/- 0.56 ml/mm Hg.min. liter; n.s.). However, the comparisons of Lp/Vp0 among the four groups (hypo(+)/DM, hypo(-)/DM, hypo(+)/non-DM and hypo(-)/non-DM) showed significant differences between hypo(+)/non-DM and hypo(-)/non-DM (1.08 +/- 0.40, 1.02 +/- 0.32, 0.71 +/- 0.29*, 1.40 +/- 0.53* ml/mm Hg.min.liter; *p < 0.05). Differences in the percentage of Vp0 to body weight (Vp0/BW) among four groups and correlation between Lp/Vp0 and Vp0/BW were not significant. CONCLUSION: These data indicated that reduction of Lp/Vp0 was not simply caused by decreased circulating plasma volume (Vp0/BW) and that the suppressed filtration coefficients may have substantial association with dialysis-induced hypotension in non-DM. The estimation of Lp using in-line measurement of Ht was a useful method for analyzing intradialytic hypotension. PMID- 12566675 TI - Can the administration of antithrombin III decrease the risk of cerebral hemorrhage in premature infants? AB - This study was carried out to determine whether the administration of antithrombin III decreases the risk of intraventricular hemorrhage in premature infants. In a randomized study, 60 infants born before 30 weeks of gestation were assigned to receive a loading dose of antithrombin III or placebo. There was no significant difference in the incidence of intraventricular hemorrhage between the antithrombin III and the placebo group (27.5 vs. 32%). Partial thromboplastin time, Quick's prothrombin time and platelet count were also not significantly different between the 2 groups. We conclude that the administration of antithrombin III during the first 2 days of life does not decrease incidence of intraventricular hemorrhage. Antithrombin III is a very expensive therapy and its benefits should be carefully investigated before being recommended as valuable therapy. PMID- 12566676 TI - Reproducibility of cerebral near infrared spectroscopy in neonates. AB - Near infrared spectroscopy (NIRS) allows to study cerebral hemodynamics and oxygenation in neonates, which may be useful for early detection of cerebral hypoxemia. So far this method is not reliable enough to be used clinically. Reproducibility is one of the prerequisites for reliable quantitative monitoring. The aim of this study was to assess the reproducibility of the NIRS parameters HbO(2) and HbD (oxygenated and deoxygenated hemoglobin concentration) and the derived NIRS parameters HbT (tissue hemoglobin concentration, HbT = HbO(2) + HbD) and rSO(2) (regional cerebral oxygen saturation, rSO(2) = HbO(2)/HbT). Two observers repeated a total number of 500 measurements in 25 neonates. Additionally, a baseline measurement was done to assess the physiological variation in every neonate. For all NIRS parameters, the inter-patient variance contributed most to the total variance, while the interobserver variance was the smallest variance component. The cerebral oxygen saturation parameter rSO(2) showed a good reproducibility, with an inter-measurement variance slightly but not significantly higher than the physiological baseline variation. The NIRS concentration parameters HbO(2), HbD, and HbT were less reproducible, with significant variation due to repeated sensor replacement. However, for cerebral oximetry rSO(2) is likely to be more important than the other NIRS parameters, so that NIRS has the potential to become a quantitative cerebral monitoring method. PMID- 12566677 TI - Cerebrovascular response in small preterm infants during routine nursery gavage feedings. AB - Although gavage feedings are considered a standard of care, they are often accompanied by hemodynamic changes that may have important effects on the cerebral circulation. In 23 premature infants receiving intermittent bolus gavage feeds, changes in cerebral hemodynamics and oxygenation were studied using near infrared spectroscopy. Orogastric tube insertion resulted in an increased cerebral blood volume in 73% of the infants and in an increase in reduced hemoglobin and in cytochrome AA(3) oxygenase in approximately 66% of the patients. Within 10 min of initiating a gavage feed, cerebral blood volume, oxygenated hemoglobin, reduced hemoglobin, and cytochrome AA(3) oxygenase decreased from baseline in about 60% of the infants. Towards the end of the study, during the postfeeding period, cytochrome AA(3) oxygenase and oxygenated hemoglobin increased in 60%, while reduced hemoglobin decreased in 78% of the infants. PMID- 12566678 TI - "Safe" hyperbilirubinemia is associated with altered neonatal behavior. AB - Treatment of neonatal jaundice is currently recommended for higher bilirubinemia levels than before. Using the Brazelton Neonatal Behavior Assessment Scale, we found that a series of 28 healthy, untreated, term neonates with moderate bilirubinemia scored significantly less than an equal number of appropriately matched controls with low bilirubinemia for visual and auditory items, both inanimate and animate. Also, a greater lability of state, a lower self-quieting ability and more frequent tremors were found in the jaundiced group. We conclude that hyperbilirubinemia per se, even in the concentration range where phototherapy is not currently recommended, can give rise to alterations in neonatal behavior. PMID- 12566679 TI - Exposure to Di(2-ethylhexyl)phthalate in humans during pregnancy. A preliminary report. AB - BACKGROUND: Di(2-ethylhexyl)phthalate (DEHP), the most commonly used plasticizer, is a widespread ubiquitous environmental contaminant. The potential health hazards from exposure to DEHP and its main metabolite, mono(2 ethylhexyl)phthalate (MEHP), have been well documented. Exposure to DEHP and MEHP in humans at risk, such as pregnant women and human fetuses, has not been tested. METHODS: Plasma DEHP and MEHP concentrations were measured in a total of 24 consecutive mother-infant pairs by high performance liquid chromatography. Associations between DEHP/MEHP and infant characteristics were tested using Fisher's exact test, unpaired t tests and univariate linear regression analysis. RESULTS: Measurable DEHP and MEHP concentrations were found in 17/24 (70.8%) and 18/24 (75%) maternal plasmas, respectively, and in 11/25 (44%) and 18/25 (72.0%) cord samples, respectively. Either DEHP or MEHP were detectable in 21/24 (87.5%) maternal plasmas and 19/25 (76%) cord samples. The mean DEHP concentrations in maternal and cord plasmas were 1.15 +/- 0.81 and 2.05 +/- 1.47 microg/ml, respectively. The mean MEHP concentrations were 0.68 +/- 0.85 and 0.68 +/- 1.03 microg/ml, respectively. No significant correlations were found between maternal and cord blood DEHP, maternal and cord blood MEHP, maternal DEHP and cord blood MEHP, or maternal MEHP and cord blood DEHP plasma concentrations. CONCLUSION: Although the effects of perinatal exposure to phthalates need further research, our findings: (i) confirm the high frequency of DEHP and/or MEHP exposure in human pregnancies; (ii) indicate that the exposure to these environmental contaminants begins during intrauterine life, and (iii) suggest that fetal exposure is closely related to the maternal exposure. PMID- 12566680 TI - Evaluation of a netilmicin-loading dose in very low birthweight infants. AB - BACKGROUND: The bactericidal efficacy of aminoglycosides is directly related to maximum serum concentrations, particularly the initial one. Therefore, several groups have recommended an aminoglycoside loading dose. Our goal was to develop a simplified dosage regimen for preterm infants which would result in therapeutic maximum serum concentrations early in the course of therapy. METHODS: Open, noncomparative study during November 2000 to April 2001. The modified netilmicin dosing protocol included a loading dose of 5 mg/kg in the first week of life, followed by a maintenance regimen of 3.5 mg/kg every 24 h. After the first week of life the corresponding doses were 6 (loading) and 5 mg/kg (maintenance). A peak level was measured 30 min after the second dose, and a trough level immediately before the third dose. RESULTS: Thirty-five very low birthweight infants (mean birthweight 876 +/- 170, range 536-1,385 g; mean gestational age 26 +/- 1.8, range 23-30 weeks) who had 46 episodes of netilmicin treatment were included in the analysis. Mean netilmicin peak and trough values were 15.9 +/- 3.7 (range 8.9-28.9) and 3.4 +/- 1.3 (range 1.0-7.8) micromol/l, respectively. Ninety-one percent of all peak levels were within the targeted range of > or =10 micromol/l. Eleven trough values (24%) were > or =4 micromol/l: in 7 instances netilmicin was administered within the first week of life, 5 of these patients had concomitant indomethacin treatment. Only 1 of the 35 neonates had a rise in serum creatinine of > or =0.5 mg/dl during netilmicin therapy. Hearing evaluations were performed in 25 of the 29 surviving infants at discharge home, all of which gave normal results. CONCLUSIONS: The new netilmicin-dosing protocol yielded therapeutic maximum serum concentrations in 91% of cases after the second dose. However, a significant number of very low birthweight infants had elevated trough levels, particularly when netilmicin was administered in the first week of life with concomitant indomethacin treatment. We speculate that a longer interval between the loading dose and the first maintenance dose would result in fewer elevated trough levels with a similarly high number of therapeutic peak levels. PMID- 12566681 TI - Early respiratory system mechanics and the prediction of chronic lung disease in ventilated preterm neonates requiring surfactant treatment. AB - The objective of this study was to evaluate the predictive value of the respiratory system compliance (Crs) and resistance (Rrs) measured before surfactant therapy to identify infants at risk for chronic lung disease (CLD). Measurements of Crs and Rrs were obtained on 44 ventilated neonates with respiratory distress syndrome (RDS) before and after surfactant therapy using the passive expiratory flow-volume method. It was found that in addition to a lower gestational age (GA) and a lower birth weight (BW), infants with CLD (n = 10) exhibited a lower Pa/AO(2) [Pa/AO(2) = PaO(2)/(PiO(2) - PaCO(2))] and higher Rrs before surfactant compared to the infants without CLD (n = 28). Improvement in gas exchange 18 h after surfactant was reduced in the CLD group. Finally, we concluded that a low GA (< 28 weeks), a low BW (< 942 g), a low Pa/AO(2) before and 18 h after surfactant or a high Rrs before surfactant (>0.21 cm H(2)O/ml/s) were associated with an increased risk of CLD. PMID- 12566682 TI - Influence of breast feeding, and adapted and hydrolyzed formulas on biliary bile acids in newborn guinea pigs. AB - BACKGROUND: The type of feeding can differently influence bile acid ontogeny during the period of physiologic cholestasis that characterizes early human development. AIM: To investigate the effects of feeding from birth, conventional or partially protein hydrolyzed cow's milk formulas were compared with breast milk regarding bile acid concentration and the composition of gallbladder bile in newborn guinea pigs. METHODS: Forty newborn guinea pigs were allocated to one of three different feeding regimens: breast milk (n = 14), intact protein formula (n = 13) and partially protein hydrolyzed formula (n = 13). After 8 days, one third of the pups from each group was sacrificed; another third was sacrificed on the 14th day and the remainder on the 20th day of life. Bile acids in gallbladder bile were analyzed by gas chromatography/mass spectrometry and HPLC. RESULTS: During the first 2 weeks of life, weight gain was significantly higher in breast fed than in artificially fed pups. An age-related increase in total biliary bile acid concentrations was evident for breast-fed and hydrolyzed formula-fed animals, but not for those fed intact protein formula. Breast-fed animals had the highest total biliary bile acid concentrations on day 20 of life, with significant increases in chenodeoxycholic and 7-oxo-lithocholic acid concentrations, which were absent in the other two groups. Concentrations of 7 oxo-lithocholic acid on day 8 were significantly higher in animals fed intact protein formula compared with breast-fed and partially protein hydrolyzed formula fed animals. CONCLUSIONS: In newborn guinea pigs, breast feeding is associated with a marked increase in biliary bile acid concentrations, which was not observed in artificially fed animals. The higher biliary bile acid concentrations and better weight gain in our breast-fed animals may reflect a greater feed efficiency associated with natural feeding. Biliary bile acid composition on day 8 suggests more rapid intestinal bacterial bile acid biotransformation in animals fed intact protein formula compared to other feeding regimens. PMID- 12566683 TI - Endogenous production of nitric oxide in endotoxemic piglets. AB - We sought to assess the relation between endotoxin-induced pulmonary hypertension and the production of nitric oxide (NO) in neonatal animals. Adult animals respond to endotoxin by increasing exhaled NO and plasma NO metabolites. The response of neonatal animals has not previously been reported. We administered 20 microg/kg of Escherichia coli lipopolysaccharide (LPS) to 12- to 18-day-old and to 5- to 7-week-old piglets. Pulmonary vascular resistance increased significantly in both age groups. Exhaled NO in the 12- to 18-day-old animals and in the 5- to 7-week-old piglets did not increase significantly. A similarly treated group of adult rats did show a significant increase in exhaled NO (2.6 +/ 1.0 to 109.5 +/- 54.3 ppb; p = 0.028). Plasma NO metabolite measurements followed the same pattern of no increase in both porcine groups, and a large increase in the rat group. However, immunostaining of lungs from 12- to 18-day old piglets did reveal an increase in inducible NO synthase. These results suggest that piglets demonstrate a limited ability to modulate LPS-induced pulmonary hypertension by elevations in exhaled NO. They also demonstrate the differential response to LPS between species. PMID- 12566684 TI - Modified protocols for surfactant therapy in experimental meconium aspiration syndrome. AB - In adult rats with experimental meconium aspiration syndrome, we investigated whether the therapeutic effect of exogenous surfactant was increased by addition of dextran or preceding airway lavage with diluted surfactant. Animals (n = 72) ventilated with pure oxygen were given human meconium suspension (50-75 mg kg( 1)) through the airways. When the PaO(2) had decreased to <20 kPa (mean +/- SD 12 +/- 3.9 kPa), the rats were randomly allocated to ten groups (G). G 6-10 underwent lung lavage with diluted Curosurf (5 mg ml(-1), 20 ml kg(-1)), whereas G 1-5 did not. G 1 and 6 received no additional material through the airways. G 2 and 7 received Curosurf (100 mg kg(-1)), and G 3 and 8 received Curosurf (100 mg kg(-1)) plus dextran (75 mg kg(-1)); G 4 and 9 received Curosurf (200 mg kg(-1)), and G 5 and G 10 received Curosurf (200 mg kg(-1)) plus dextran (75 mg kg(-1)). All rats in G 1 died before 180 min after randomization. In G 2, 3, 6, 7, and 8, the PaO(2) transiently increased to 30-40 kPa. In G 4, 5, 9, and 10, the PaO(2) remained >30 kPa for 180 min. Both airway lavage and supplementation with dextran improved the therapeutic effects of surfactant; however, a large dose (200 mg kg( 1)) was nevertheless required to optimize gas exchange. PMID- 12566685 TI - Geometric properties of the lung parenchyma after postnatal glucocorticoid treatment in rats. AB - While glucocorticoid (GC) administration appears to be beneficial during the acute phase of treatment of neonates at risk of developing chronic lung disease, it is still not clear whether steroid application has an adverse long-term effect on the lung maturation. Thus, the goal of the present work was to analyze GC effects on the pulmonary structure in a rat model where dosage and timing of drug administration were adapted to the therapeutic situation in human neonatology. The animals received daily a maximum of 0.1 mg dexamethasone phosphate per kilogram body weight during the first 4 postnatal days. Investigations were performed at the light microscopic level by means of a digital image analysis system. While there were no differences in the lung architecture between experimental animals and controls on day 4, the earliest time point of observation, we found a widening of airspaces with a concomitant decrease in the alveolar surface area density, representing a loss of parenchymal complexity, on days 10 and 21 in treated rats. On days 36 and 60, however, no alterations in the pulmonary parenchyma could be detected in experimental animals. We conclude from these findings that the GC-induced initial inhibition of development (days 10 and 21) was completely reversed, so that a normal parenchymal architecture and also a normal alveolar surface area density were found in adult rats (days 36 and 60). From the results obtained using the regimen of GC administration described, mimicking more closely the steroid treatment in human neonatology, we conclude that the observed short-term adverse effects on lung development can be fully compensated until adult age. PMID- 12566686 TI - Pitfall of newborn screening for congenital adrenal hyperplasia due to 21 hydroxylase deficiency. AB - We report on a newborn with a severe salt-wasting crisis due to congenital adrenal hyperplasia (CAH) with 21-hydroxylase deficiency, in spite of prenatal diagnosis of CAH and awareness of this diagnosis by the parents and the gynecologic and pediatric practitioners. Due to the diagnosis of CAH in the older sister, prenatal treatment with dexamethasone (Dexa) was initiated. Prenatal diagnosis showed an affected male fetus and the prenatal treatment was stopped. The parents and the involved physicians were informed about the diagnosis, treatment, follow-up, and possible complications. Amniotic infection led to preterm delivery in another hospital. Due to perinatal asphyxia, the male newborn received Dexa during mechanical ventilation. Neonatal CAH screening was unsuspicious. An acute salt-wasting crisis with metabolic acidosis at the age of 3 weeks finally led to the correct diagnosis. This experience emphasizes the need to obtain a careful medical history. Furthermore, this case illustrates that neonatal screening for CAH is falsely negative in the event of neonatal Dexa treatment. PMID- 12566687 TI - A rare case of neonatal ileo-cecal valve stenosis due to covered iliac perforation. AB - At birth, newborn babies tend to respond to any stressful event with elective blood shunting towards the main organ systems, such as the brain and heart. Consequently, the bowel may suffer from a severe flow reduction with a high risk of hypoperfusion. The distal ileum is relatively less vascularized than other parts of intestine, due to a lack of collateral arteriolar circulation. It is therefore at higher risk of hypoxia, tissue necrosis and perforation in situations of minimal hypoperfusion or spasm. A rare case of covered perforation of the last loop of the ileum in a severely preterm baby is reported. The perforation manifested atypically as inflammatory stenosis of the loop and the ileo-cecal valve. To avoid misdiagnoses and to manage these cases correctly, the caregiver has to consider this rare eventuality in the differential diagnosis of intestinal occlusion/subocclusion in the first days of life. PMID- 12566688 TI - Relationships between human auditory cortical structure and function. AB - The human auditory cortex comprises multiple areas, largely distributed across the supratemporal plane, but the precise number and configuration of auditory areas and their functional significance have not yet been clearly established. In this paper, we discuss recent research concerning architectonic and functional organisation within the human auditory cortex, as well as architectonic and neurophysiological studies in non-human species, which can provide a broad conceptual framework for interpreting functional specialisation in humans. We review the pattern in human auditory cortex of the functional responses to various acoustic cues, such as frequency, pitch, sound level, temporal variation, motion and spatial location, and we discuss their correspondence to what is known about the organisation of the auditory cortex in other primates. There is some neuroimaging evidence of multiple tonotopically organised fields in humans and of functional specialisations of the fields in the processing of different sound features. It is thought that the primary area, on Heschl's gyrus, may have a larger involvement in processing basic sound features, such as frequency and level, and that posterior non-primary areas on the planum temporale may play a larger role in processing more spectrotemporally complex sounds. Ways in which current knowledge of auditory cortical organisation and different data analysis approaches may benefit future functional neuroimaging studies which seek to link auditory cortical structure and function are discussed. PMID- 12566689 TI - High-frequency sensitivity of the mature gerbil cochlea and its development. AB - The thresholds of compound action potentials evoked by tone pips were measured in the cochleae of anesthetized gerbils, both in adults and in neonates aged 14, 16, 18, 20 and 30 days, using round-window electrodes. Stapes vibrations were also measured, using a laser velocimeter, in many of the same ears of adults and neonates aged 14, 16, 18 and 20 days to assess cochlear sensitivity in isolation from middle ear effects and to circumvent problems associated with calibration of acoustic stimuli at high frequencies. Whether referenced to sound pressure level in the ear canal or stapes vibration velocity, thresholds in adults were roughly uniform in the entire range of tested frequencies, 1.25-38.5 kHz. In neonates, thresholds decreased systematically as a function of age, with the largest reductions occurring at the highest frequencies. Thresholds remained slightly immature at all frequencies 30 days after birth. The results for adult gerbils are consistent with the recent finding that basilar-membrane responses to characteristic frequency tones normalized to stapes vibrations are as sensitive at sites near the round window as at more apical sites. The results for neonates confirm that the extreme basal region of the cochlea is the last to approach maturity, with substantial development occurring between 20 and 30 days after birth. PMID- 12566690 TI - Expression of the P2X7 receptor subunit of the adenosine 5'-triphosphate-gated ion channel in the developing and adult rat cochlea. AB - ATP-gated ion channels assembled from P2X(7) subunits have been implicated in ontogeny and cellular pathology. Here, the expression of the P2X(7) receptor subunit was studied in the embryonic (E14-E18 days) and postnatal (P0-adult) rat cochlea using immunohistochemistry. Strong P2X(7) immunolabelling was observed in the primary auditory neurons of the spiral ganglion from E18 to adult and in the fibres innervating the sensory inner and outer hair cells from birth to adult. Strong immunolabelling of P2X(7) receptor protein was also observed in the inner and outer hair cells over a limited developmental period, from birth to P6. Weak expression was observed in cochlear duct epithelium on E18 and in the supporting cells (footplates of pillar cells in adult and in Bottcher's cells after birth). The immunolocalisation of P2X(7) receptors further implicates extracellular ATP in signalling process in cochlear ontogeny and in establishment and function of auditory neurotransmission. The P2X(7) receptors may be involved in signal transduction and modulation as well as in regulating cell death during development and in pathological conditions. PMID- 12566691 TI - Gentamicin-induced cytotoxicity involves protein kinase C activation, glutathione extrusion and malondialdehyde production in an immortalized cell line from the organ of corti. AB - The objective of the present study was to investigate the biochemical mechanisms underlying gentamicin cytotoxicity in OC-k3 cells derived from an immortalized cell line developed from the organ of Corti of transgenic mice. Administration of 50 microM gentamicin significantly reduced cell proliferation and viability, as well as initiating morphological changes associated with apoptosis. Protein kinase C (PKC) alpha activity was increased in gentamicin-treated cells, peaking 15 min after dosing (+138.2%). This PKCalpha increase was followed by a rise of glutathione (GSH) efflux and a concomitant 29% decrease in intracellular GSH levels at 30 min. PKCalpha-specific inhibitors blocked these cytotoxic effects. Gentamicin also increased malondialdehyde levels, while N-acetylcysteine, GSH and ascorbic acid prevented gentamicin-induced cell death. These findings suggest that gentamicin cytotoxicity involves a production of intracellular reactive oxygen species and a concomitant PKC-dependent fall of intracellular scavenging abilities (GSH), events that together drive cells to undergo apoptosis. PMID- 12566692 TI - Pharmacokinetics of caroverine in the inner ear and its effects on cochlear function after systemic and local administrations in Guinea pigs. AB - Caroverine, an N-methyl-D-aspartate and alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid receptor antagonist, has been shown to protect the inner ear from excitotoxicity and to be effective in the treatment of cochlear synaptic tinnitus. Local administration of caroverine on the round window membrane (RWM) could be a more effective means of administration to avoid systemic side/adverse effects. The present study investigates the pharmacokinetics of caroverine in the perilymph, cerebrospinal fluid (CSF) and plasma following intravenous and local applications at different dosages. High-performance liquid chromatography was used to determine the drug concentrations. Our results show much higher caroverine concentrations in the perilymph with lower concentrations in CSF and plasma following local applications compared with systemic administration. Auditory brainstem responses were measured to evaluate the changes in auditory function. The effects on hearing were transient and fully reversible 24 h after local caroverine applications. The findings suggest that local application of caroverine on the RWM for the treatment of excitotoxicity-related inner ear diseases, such as tinnitus and noise-induced hearing loss, might be both safe and more efficacious while avoiding high blood and CSF caroverine levels seen with systemic administration. PMID- 12566693 TI - Tubular and interstitial expression of ICAM-1 as a marker of renal injury in IgA nephropathy. AB - BACKGROUND: The upregulated renal expression of intercellular adhesion molecule 1 (ICAM-1) is associated with glomerular and interstitial infiltration of leukocytes. AIM: To test the hypothesis that renal expression of ICAM-1 may be predictive in the highly variable IgA nephropathy (IgAN). METHODS: ICAM-1 (CD54) in tubular epithelium and interstitial leukocytes, macrophages (CD14), and T cells (CD3) were assessed using avidin-biotin-peroxidase in renal biopsy specimens from 45 patients with IgAN and from 29 patients with no glomerulonephritis. RESULTS: In IgAN, tubular ICAM-1+ was seen in 25 of 45 (55%) biopsy specimens, associated with glomerular hypercellularity, glomerulosclerosis involving less than 50% of the glomerular area, interstitial cellular infiltration, tubular atrophy, and proteinuria (U = 44, p = 0.005). Interstitial ICAM-1+ leukocytes were correlated with glomerulosclerosis involving less and more than 50% of the glomerular area, tubular atrophy, interstitial fibrosis, and serum creatinine concentration (r = 0.6343, p < 0.001). In patients with an increase of 50% in the serum creatinine concentration, interstitial ICAM-1+ leukocytes and CD14+ and CD3+ cells were significantly more numerous than in patients with a stable creatinine concentration. In patients with no glomerulonephritis, tubular ICAM-1+ was seen in 7 of 29 (24%) biopsy specimens, inversely correlated with the number of normal glomeruli and associated with glomerulosclerosis covering more than 50% of the glomerular area, tubular atrophy, and creatinine. CONCLUSIONS: Tubular and interstitial expression of ICAM 1 can be a marker of tubulointerstitial disturbance in IgAN. Interstitial ICAM-1 may be an adverse predictor of disease progression. PMID- 12566694 TI - A point mutation in the first splice donor leads to reduced oncogenic properties of the adenovirus serotype 12 E1A gene. AB - Cells transformed by proteins of early regions 1A (E1A) and 1B (E1B) of oncogenic adenovirus serotype 12 (Ad12) grow to tumours in syngeneic, immunocompetent rodents. To gain insight into the mechanisms of oncogenic transformation, we point mutated the first splice donor in the Ad12-E1A gene, leading to the loss of the Ad12-E1A(9.5S) and Ad12-E1A(11S/10S) proteins and to a conservative amino acid (aa) exchange at position aa 30 (valine vs. leucine) in the Ad12-E1A(13S) and Ad12-E1A(12S ) proteins. BMK cells transformed by mutant Ad12-E1A (Ad12-E1Am) plus Ad12-E1B via retrovirus-mediated gene transfer showed features comparable to wild-type Ad12-E1A (Ad12-E1Awt) plus Ad12-E1B-transformed cells: they formed foci in soft agar and produced tumours in immunodeficient nude mice, although after a prolonged latency period. These results suggest that Ad12-E1A(9.5S) and Ad12 E1A(11S/10S) are dispensable for cellular transformation. However, in contrast to Ad12-E1Awt cells, Ad12-E1Am cells failed to grow to tumours in syngeneic, immunocompetent rodents, with the exception of one cell line, which produced tumours in about 50% of the immunocompetent animals. Interestingly, the concentration of the putative tumour suppressor and co-activator p300 was elevated in cell lines expressing high levels of Ad12-E1A and Ad12-E1B due to an increased half-life. These results indicate that p300 is stabilized in Ad12-E1 transformed BMK cells, probably by a mechanism linked to high expression of Ad12 E1A/E1B. PMID- 12566695 TI - Differences in release and determination of subtype of porcine endogenous retroviruses produced by stimulated normal pig blood cells. AB - OBJECTIVE: Porcine endogenous retroviruses (PERVs) are of particular concern with xenotransplantations using pig cells, tissues or organs as they are present in the genome of all pig strains and are able to infect human cells in vitro. However, it remains unclear whether PERV particles will be produced in vivo and whether they may infect xenotransplant recipients. Since normal pig peripheral blood mononuclear cells (PBMCs) may be transmitted together with the transplanted organ, the production of PERVs by stimulated PBMCs was studied in vitro. METHODS: To simulate antigen-induced activation of PBMCs, phytohaemagglutinin (PHA), a T cell mitogen, and the phorbol ester O-tetradecanoylphorbol-13-acetate (TPA), a tumour promoter, were used. Virus release was estimated by measuring reverse transcriptase (RT) activity and by RT-PCR of pelleted viruses. RESULTS: Treatment of pig PBMCs with PHA or TPA induced the release of PERVs. For the first time, a correlation between the extent of proliferation of pig PBMCs and PERV production was shown. In addition, PERV release by non-proliferating cells and differences in virus production between stimuli as well as between different pig strains and individuals of one strain were observed. Subtype analysis revealed the release of the three subtypes PERV-A, PERV-B and PERV-C. In contrast to murine endogenous retroviruses, PERVs were induced by PHA alone. CONCLUSION: The data suggest that the PBMCs transmitted within a xenotransplant may release PERV. These data also suggest that pig strains producing low amounts of virus could be more suitable for xenotransplantation. PMID- 12566696 TI - Transcription at early stages of herpes simplex virus 1 infection and during reactivation. AB - OBJECTIVE: The kinetics of immediate early (IE) and early (E) herpes simplex virus 1 (HSV-1) mRNA transcription was followed in explanted trigeminal ganglia from rabbits with established latency. METHODS: The expression of IE and E mRNAs was first assessed in infected Vero cells by RT-PCR and then in explanted trigeminal ganglia by nested RT-PCR. RESULTS: In infected Vero cells, IE mRNAs [for infected cell protein (ICP) 0, ICP4 and ICP27] were first detected 1-2 h post-inoculation (p.i.), peaking at 3 h p.i. The transcription of E mRNAs [for thymidine kinase (TK), RR1 and UL9], which were first detected from 3 h p.i., peaked between 5 and 10 h p.i. In explanted ganglia, the ICP0, ICP4 and ICP27 mRNAs were first detected after 4 h in culture. This was followed by the appearance of TK mRNA at 8 h and then by the UL9 mRNA, detected from 12 h post explantation. A further E mRNA (RR1), as well as the late gC mRNA, were first observed after 24 h in culture. Moreover, ICP4 mRNA could be found in non cultured ganglia. CONCLUSIONS: During reactivation of latent HSV-1 in explanted ganglia, the onset of ICP0 and ICP27 transcription at 4 h in culture was followed by TK transcription (at 8 h). Thus, in the rabbit reactivation model, ICP0 gene transcription rather than ICP4 transcription represents the relevant indicator of latency reactivation. PMID- 12566697 TI - Circulating intercellular adhesion molecule-1 in chronic hepatitis C patients with normal or elevated aminotransferase before and after alpha-interferon treatment. AB - OBJECTIVES: Intercellular adhesion molecule-1 (ICAM-1) plays a fundamental role during liver inflammation. In fact, weak ICAM-1 expression is physiologically restricted to the endothelium of portal vessels and to sinusoidal lining cells, but it becomes markedly evident on sinusoidal lining cells and at the surface of hepatocytes during inflammatory liver diseases. The aim of this study was to evaluate the behaviour of soluble ICAM-1 (sICAM-1) in chronic hepatitis C (CH-C) patients with persistently normal aminotransferase in comparison with patients with CH-C and elevated aminotransferase, and its changes during alpha-interferon (IFN) therapy. Immunohistochemical localization of ICAM-1 was also performed on liver tissue specimens of both groups. METHODS: Sixty subjects were divided into 3 groups: group A included 19 patients with CH-C and persistently normal aminotransferase; group B included 21 patients with CH-C and persistently elevated aminotransferase levels, and group C included 20 healthy subjects representing the control group. The first two groups were treated with recombinant alpha-IFN 2b at a dose of 6 MU 3 times a week for 3 months and followed up with 3 MU 3 times a week for another 3 months. RESULTS: Baseline values of serum ICAM-1 in groups A and B were significantly higher than those in group C (p < 0.0001). The median baseline value of sICAM-1 in group A (525.0 ng/ml) was lower than that of group B (561.0 ng/ml), but the difference was not statistically significant. Furthermore, there was a trend toward higher ICAM-1 values as histological severity increased (Mantel-Haenszel chi(2) 8.8, p < 0.003). Post-treatment sICAM-1 serum values showed a marked decrease in both groups, but only among responder patients, while ICAM-1 levels were unchanged in non-responders. Immunohistochemical localization showed no staining for ICAM-1 in normal liver specimens, while there was a quite similar staining for ICAM-1 in the two groups of patients, consistent with an inflammatory process. CONCLUSIONS: This study shows that circulating ICAM-1 levels in most patients with CH-C and persistently normal aminotransferase are higher than those in a control group and the fact that ICAM-1 molecules are also expressed on the hepatocyte membrane suggests that they could play an important role, in association with other molecules, in the intercellular adhesion processes during the induction and maintenance phases of the immune response. In both groups, only patients responding to alpha-IFN therapy showed a marked decrease in serum ICAM-1 below baseline values. PMID- 12566698 TI - Hepatitis B virus DNA in anti-HBe-positive asymptomatic carriers. AB - OBJECTIVES: We examined the level of hepatitis B virus (HBV) DNA and the mutations in the precore, core and polymerase regions of HBV in sera from anti HBe-positive asymptomatic carriers (ASC). METHODS: The amount of HBV DNA was determined semiquantitatively by mutation site-specific assay in sera from 19 anti-HBe-positive ASC and 31 HBeAg-positive patients with chronic liver disease (CLD). The mutations in the precore, core and polymerase (terminal protein) regions, spanning 1,037 nucleotides, of HBV in sera from three cases each of anti HBe-positive ASC, HBeAg-positive ASC and HBeAg-positive CLD were examined by directly sequencing the amplified HBV DNA. RESULTS: The level of HBV viremia in anti-HBe-positive ASC was significantly lower than that in HBeAg-positive CLD patients (p < 0.01). By sequence analysis, there were a few missense mutations detected in HBeAg-positive ASC and HBeAg-positive CLD patients. In contrast, many mutations, especially in the central or N-terminal half of the core region and N terminal part of the polymerase region, were detected in anti-HBe-positive ASC. CONCLUSION: Mutations not only in the precore/core region, but also in the polymerase region of HBV might cause damage to some important functions for efficient replication of HBV and be involved in the reduced amount of HBV in anti HBe-positive ASC. PMID- 12566699 TI - Analysis of saturation binding and saturation infection for dengue serotypes 1 and 2 in liver cells. AB - OBJECTIVE: The liver has been increasingly recognized as a significant target organ in the pathogenesis of dengue virus infection. However, only two contradictory studies have examined the binding of the dengue virus to liver cells. This study therefore sought to investigate the binding of the dengue virus to HepG2 cells. METHODS: Radiolabeled dengue virus serotypes 1 and 2 were prepared through viral propagation in Vero cells. Increasing amounts of virus were then incubated with HepG2 cells to determine the ability of the virus to achieve saturation of binding on HepG2 cells. RESULTS: Results indicated that it was not possible to reach saturation of binding under experimentally achievable conditions. We then sought to determine whether it was possible to reach a state of saturation of infection, by using increasingly high titers of virus on a constant number of cells. Dengue serotype 1 showed no evidence of saturation of infection, even at titers of 5,000 viruses per cell. In contrast, dengue serotype 2 became saturated at levels of approximately 3,000 viruses per cell. CONCLUSIONS: These results are consistent with proposals that dengue virus binding to cells is mediated initially through a low-affinity interaction with an abundant molecule on the surface of the cell and secondly through interaction with a less commonly expressed molecule, which is required for viral internalization. PMID- 12566700 TI - Autoantibodies to CD69 in patients with chronic hepatitis type C: a candidate marker for predicting the response to interferon therapy. AB - OBJECTIVE: To understand the autoimmunity associated with chronic hepatitis C (CHC), we investigated autoantibodies (autoAbs) to CD69. METHODS: With this aim, we tested the reactivity of serum samples from patients with CHC and asymptomatic carriers of hepatitis C virus (HCV), as well as from patients with chronic hepatitis B (CHB) and autoimmune hepatitis (AIH), to recombinant CD69 molecules. RESULTS: Frequencies of anti-CD69 autoAbs were 38.7% in CHC, 15.8% in AIH and 12.3% in CHB. None of the tested asymptomatic HCV carriers had autoAbs to CD69. It is important clinically that the presence of anti-CD69 autoAbs was found to be associated with a poor response to interferon-alpha (IFN-alpha) therapy. In the epitope analysis, multiple epitopes were mapped on CD69, indicating antigen driven production of the autoAbs. CONCLUSION: We evidenced existence of anti-CD69 autoAbs in patients with CHC, and found that the anti-CD69 autoAb may have potential for predicting responses to IFN-alpha therapy. PMID- 12566701 TI - Genetic characterization of a porcine H1N2 influenza virus strain isolated in Germany. AB - We examined selected influenza virus strains from various outbreaks of respiratory diseases among pigs in Germany. Besides H3N2 and H1N1 subtypes presently circulating in Europe, an H1N2 influenza virus was isolated in 2000. Sequence analysis of hemagglutinin, neuraminidase and matrix genes indicated that this virus is a reassortant of human H1N1 and human H3N2 strains with an internal gene, the M gene of avian origin. It is a result of a multiple reassortment event. PMID- 12566702 TI - Overview of the needs and realities for developing new and improved vaccines in the 21st century. AB - The science of present day vaccinology is based on the pioneering discoveries of the late 18th and late 19th centuries and the technologic breakthroughs of the past 60 years. The driving force for the development of new vaccines resides in technologic feasibility, public need and economic incentive for translating the basic knowledge into a product. Past efforts by government to define which particular vaccines to develop were mostly irrelevant to the realistic choices which were made. There is a vast array of viral, bacterial, parasitic and fungal disease agents against which preventative vaccines may be developed, and to this may be added cancer and certain amyloidoses such as Alzheimer's and 'mad cow' diseases. The proven past for vaccines has relied on live, killed, protein and polysaccharide antigens plus the single example of recombinant-expressed hepatitis B vaccine. The validity of redirection of vaccinology to exploration of simplified vaccines such as recombinant vectored and DNA preparations and reductionist vaccines based on peptides of contrived epitope composition remains to be proved. Reductionism imposes vastly increased complexity and difficulty on vaccine development and might not be capable of achievement. The challenge in the 21st century will involve new and uncertain pathways toward worthwhile accomplishments. PMID- 12566703 TI - Virus-like particles as a modular system for novel vaccines. AB - Induction of protective immune responses with recombinant antigens is a major challenge for the vaccine industry. Here we present a molecular assembly system that renders antigens of choice highly repetitive. Using this method, efficient antibody responses may be induced in the absence of adjuvants resulting in protection from viral infection and allergic reactions. PMID- 12566704 TI - Molecular epidemiology and immunology of hepatitis B virus infection - an update. AB - Hepatitis B virus (HBV) continues to be one of the most important viral pathogens in humans. This review provides an update on the molecular epidemiology and immunology of HBV infection. DNA sequencing has allowed replacement of the initial serotypic classification of HBV strains by a more systematic genotype system that currently consists of 7 members (genotypes A-G). More recently, sequence analysis of virus isolates from many individual patients has revealed the occurrence of certain mutational hot spots in the genome, some of which appear to correlate with the patient's immunological and/or disease status; however, cause and effect are not always easily discernible. This holds particularly for the issue of whether virus variants exist that have, per se, an increased pathogenic potential; due to the scarcity of appropriate experimental in vivo models, such hypotheses are difficult to prove. Similarly, because of the compact organization of the HBV genome, almost every single mutation may have pleiotropic phenotypic effects. Nonetheless, there is accumulating evidence that at least some frequently observed mutations are causally related to viral escape from selective pressures, such as the presence of antibodies against dominant B cell epitopes, or drugs that inhibit the viral reverse transcriptase; possibly, this is also true for the cellular immune response. Therefore, despite the availability of an effective prophylactic vaccine, further extensive efforts are required to monitor the emergence of vaccination- and therapy-resistant HBV variants and to prevent their spread in the general population. PMID- 12566705 TI - In vitro expression of human hepatitis B virus genomes carrying woodchuck hepatitis virus pre-S1 sequences. AB - Many hepatitis B virus (HBV) in vivo experiments are unfeasible because this virus infects only humans. Former studies demonstrated that the pre-S1 domain of the viral envelope protein L determines host range. Therefore, we tried to generate HBV recombinants which might be able to infect woodchucks by exchanging different portions of the pre-S1 encoding gene with homologous parts from the related woodchuck hepatitis virus (WHV) in a cloned HBV genome. In 6 mutants, 11 92 N-terminal HBV pre-S1 codons were replaced by 20-120 codons from WHV. Four mutants carried C-terminal substitutions. The pre-S1 region overlaps with the viral polymerase gene, which is therefore also affected. After transfection of Huh7 cells, the DNA polymerase activity in cytoplasmic nucleocapsids was found to be only slightly affected. All mutants except for the largest C-terminal substitution allowed virion formation. Only the smallest N- and C-terminal substitutions had a wild-type phenotype. The remaining 7 variants allowed virion yields between 5 and 50% of that of the wild type. This demonstrated that substitution of up to 92 pre-S1 codons in an HBV genome with up to 120 codons from WHV pre- S1 was compatible with DNA replication and virion formation. Some recombinant viruses might be able to grow in woodchucks. PMID- 12566706 TI - Immunogenic effects of woodchuck hepatitis virus surface antigen vaccine in combination with antiviral therapy: breaking of humoral and cellular immune tolerance in chronic woodchuck hepatitis virus infection. AB - OBJECTIVE: A rational treatment strategy for chronic hepatitis B virus (HBV) infection might involve the modulation of immunity after the reduction of viremia and antigenemia. This strategy was tested in woodchucks chronically infected with the woodchuck hepatitis virus (WHV) by combining antiviral treatment with 1-(2 fluoro-5-methyl-beta-L-arabinofuranosyl)-uracil (L-FMAU) and therapeutic vaccination with WHV surface antigen (WHsAg). METHODS: Chronic WHV carriers were treated with L-FMAU or placebo for 32 weeks. Half the woodchucks in each group then received four injections of a conventional WHsAg vaccine during the next 16 weeks. RESULTS: Vaccination alone elicited low-level antibody to WHsAg (anti-WHs) in most carriers but did not affect serum WHV DNA, WHsAg or liver enzyme responses. Carriers treated first with L-FMAU to reduce WHV DNA and WHsAg and then vaccinated developed similar low-level anti-WHs and normalized liver enzymes. Following vaccinations, WHsAg-specific cell-mediated immunity (CMI) was demonstrated in both groups, but was significantly enhanced in carriers treated with L-FMAU, and was broadened to include WHV core antigen (WHcAg) and selected peptide epitopes of WHcAg and WHsAg. Anti-WHs and associated CMI to WHcAg and WHsAg were observed after drug discontinuation in half of the carriers that received L-FMAU alone. CONCLUSIONS: Vaccination with WHsAg following treatment with L-FMAU disrupted virus-specific humoral and cell-mediated immune tolerance in chronic WHV infection and enhanced the immune response profiles beyond those seen with monotherapies alone. The combination therapy resulted in immune response profiles that resembled those observed during resolution of WHV infection. The results in woodchucks demonstrate the feasibility of using such a combination therapy for the control of chronic HBV infection in humans. PMID- 12566707 TI - Selective expression of immunogenic, virus-like particle-derived antibody-binding epitopes. AB - The incorporation of linear and conformational antibody-binding epitopes into polyepitope, chimeric antigens with satisfactory immunogenicity is a challenge. We selectively expressed antigen fragments encoding the linear e2 epitope (C(79 149)) of hepatitis B virus (pre)core antigen (HBc/eAg) and the conformational 'a' epitope (S(80-180)) of hepatitis B surface antigen (HBsAg) in a novel system. The domains were expressed as chimeric antigens containing either heat shock protein (hsp)73-binding simian virus 40 large tumor antigen (e.g. T(77)) or non-hsp binding (e.g. T(60)) sequences at their N-termini. We compared their type of expression with their immunogenicity for B cells (when delivered as a DNA vaccine). The type of expression investigated included their level of expression, the secretion or intracellular expression of the antigen and the stress protein (hsp)-associated versus nonassociated expression. The linear e2 epitope of HBc/eAg was efficiently expressed as an intracellular, hsp73-binding fusion protein, and efficiently primed an HBc/eAg-specific antibody response when delivered in this form. The conformational 'a' epitope of HBsAg most efficiently stimulated B cells as a secreted, non-hsp-associated fusion protein. These data demonstrate that different B cell-stimulating epitopes of vaccine-relevant viral antigens can be selectively isolated and expressed in suitable expression systems, but that the requirements that have to be fulfilled to obtain optimal immunogenicity differ strikingly between individual epitopes. PMID- 12566708 TI - HIV: epidemiology and strategies for therapy and vaccination. AB - In the decades since its discovery, the acquired immunodeficiency syndrome AIDS has developed into one of the leading causes of death worldwide, with a dramatic situation in many countries of subsaharan Africa. A large body of knowledge has been acquired regarding the molecular biology and replication properties of the causative agent of AIDS, the human immunodeficiency virus (HIV). However, despite these advances the global spread of HIV and especially its spread in developing countries continues almost unabated. During the last decade, drugs inhibiting HIV replication have been introduced into clinical use and highly active antiretroviral therapy (HAART) using a combination of several drugs has been proven to slow or halt the course of the disease in most patients. However, HAART is currently only available in countries of the developed world and the emergence of resistant variants may rapidly lead to a partial loss of this therapeutic arsenal. Thus, the development of a prophylactic vaccine against HIV must be the ultimate goal to control the global pandemic of AIDS. This minireview summarises the state of the epidemic and discusses current treatment strategies as well as future developments. Finally, strategies towards the development of a vaccine against HIV are presented and recent studies using prime-boost regimens to achieve protection from disease in animal models while not preventing infection with the challenge virus are discussed. PMID- 12566709 TI - Vaccine development using the simian immunodeficiency virus model for AIDS. AB - OBJECTIVE: A number of trials in primates using a wide range of putative vaccines based on simian immunodeficiency virus (SIV) have been performed and are summarised here. METHODS: Rhesus macaques and African green monkey (AGMs) were immunised with the test vaccines and challenged with live virus to test the efficacy of the induced or transferred immune responses to protect from infection or disease development. RESULTS: In initial studies, successful protection from challenge by whole inactivated virus vaccines was subsequently shown to be mediated by immune responses to human cell rather than viral proteins. Passive transfer of neutralising antibodies failed to protect against challenge. The induction of SIV-specific cytotoxic T lymphocytes (CTLs) using lipopeptides also failed to protect from infection, and whereas the frequency of post-infection CTLs (as measured by limiting dilution CTL assay and MHC/tetramer staining) correlated inversely with the cell-associated virus load, no correlation with the plasma virus load was observed. No immunological correlation of protection could be identified in macaques immunised with live attenuated SIV, with sterilising immunity being induced as early as 10 weeks after infection with the attenuated virus. Similarly, whole inactivated virus and passive IgG transfer failed to protect the natural host AGM species from challenge with apathogenic SIVagm, although live attenuated SIVagm afforded some protection despite the lack of overt vaccine virus replication. CONCLUSIONS: 'Traditional' types of vaccine are either ineffective or inappropriate for use in humans. Current efforts are therefore focusing on the rapidly evolving field of genetic vaccines based on vector DNA and recombinant, attenuated viral and bacterial vectors. PMID- 12566710 TI - Impact of codon usage modification on T cell immunogenicity and longevity of HIV 1 gag-specific DNA vaccines. AB - In this study, we analyzed the in vitro expression, potency and longevity of immune responses induced in a Balb/c mouse model by a synthetic HIV-1 GAG gene exhibiting a codon usage that was adapted to that of highly expressed mammalian genes (syngag). In contrast to a vector containing the wild-type (wt) GAG gene, the syngag construct enabled highly efficient Gag expression in both human and rodent cell lines in complete absence of Rev and Rev-responsive element. Immunization of Balb/c mice with the wt gag plasmid DNA induced only weak and inconsistent humoral immune responses. Mice vaccinated by syngag but not wt gag developed substantial and highly consistent Gag-specific antibody titers showing a clear T helper 1 polarization even with low doses of DNA. Moreover, vaccinated mice developed a strong Gag-specific cellular immune response, including cytotoxic T cells, which was not observed in wt gag-immunized animals. Both humoral and cellular immunity were efficient and lasted for more than 20 weeks. Furthermore, the induction of the humoral as well as the cellular immune response was independent of the immunization route (intramuscular or subcutaneous). These results clearly show the advantages of codon-optimized genes with respect to the expression and immunogenicity of plasmid DNA constructs, making them promising vaccine candidates for further studies. PMID- 12566711 TI - A short introduction to papillomavirus biology. AB - In this report, the tropism of papillomaviruses, the structure of virions, the function of viral proteins and the use of pseudovirions for the analysis of the immune response against papillomaviruses and the search for the viral receptor are briefly described. PMID- 12566712 TI - T cell response to human papillomavirus 16 E7 in mice: comparison of Cr release assay, intracellular IFN-gamma production, ELISPOT and tetramer staining. AB - Successful vaccination against infections by high-risk papillomaviruses aiming at the prevention of cervical cancer most likely requires the induction of neutralizing antibodies and human papillomavirus (HPV)-specific T cells directed against early viral proteins such as E7. Whereas the technology for detection of antibodies is well established, measurement of T cells is more cumbersome and standardization of assays is difficult. By using chromium release assay, ELISPOT, tetramer staining and intracellular IFN-gamma assay, we compared the levels of HPV 16 E7-specific T cells obtained after immunization of C57BL/6 mice with different DNA expression vectors. We found that all four assays gave highly comparable results. ELISPOT can be recommended for future studies as it indicates the presence of activated (i.e. IFN-gamma-secreting) T cells in a quantitative manner and combines high sensitivity with relatively low T cell demand. PMID- 12566713 TI - Comparison of human papillomavirus type 16 L1 chimeric virus-like particles versus L1/L2 chimeric virus-like particles in tumor prevention. AB - Chimeric human papillomavirus (HPV) virus-like particles (cVLPs) with the HPV16 E7 antigen fused to either the major capsid protein, L1, or the minor capsid protein, L2, have been used independently to protect against the formation of HPV induced tumors in animal models. However, the advantages and disadvantages of both types of particles with respect to production and vaccine efficacy have never been analyzed. Therefore, in this study, we compared cVLPs with the HPV16 E7 antigen fused to L1 versus cVLPs with E7 fused to L2 with respect to their ability to protect mice from tumor challenge. The first 57 amino acids of E7 were used to overcome the size limitation and limited VLP production imposed by inserting polypeptides into L1 cVLPs. C57BL/6 mice were immunized with the above cVLPs at various doses. Tumor challenge was then performed with HPV16 E7-positive TC-1 cells. HPV16 L1-E7((1-57)) was superior to HPV16 L1/L2-E7((1-57)) in eliciting tumor protection at equivalent doses, although both types of particles were able to protect mice. Both cVLPs induced a specific cytotoxic T lymphocyte (CTL) response to the H2-D(b)-restricted E7 peptide (E7(49-57)) as determined by an ELISPOT assay and tetramer staining; however, immunization with the L1-E7((1 57)) cVLPs resulted in twofold higher CTL precursor frequencies. Our results demonstrate that cVLPs with the antigen fused to L1 are a more efficient vaccine with respect to tumor prevention than cVLPs with the antigen fused to L2. At the same time, however, L1 cVLPs are limited by the size of the antigen that can be incorporated and in the amount of cVLP that can be obtained from cultures when compared to L1/L2 cVLPs. This balances out their superior ability to induce protective immunity. PMID- 12566714 TI - Radiation-induced growth hormone deficiency. AB - Deficiency of one or more anterior pituitary hormones may follow treatment with external irradiation when the hypothalamic-pituitary axis falls within the fields of irradiation. Hypopituitarism occurs in patients who receive radiation therapy for pituitary tumours, nasopharyngeal cancer and primary brain tumours, as well as in children who undergo prophylactic cranial irradiation for acute lymphoblastic leukaemia, or total body irradiation for a variety of tumours and other diseases. The degree of pituitary hormonal deficit is related to the radiation dose received by the hypothalamic-pituitary axis. Thus, after lower radiation doses isolated growth hormone deficiency ensues, whilst higher doses may produce hypopituitarism. The timing of onset of the radiation-induced pituitary hormone deficit is also dose-dependent. The main site of radiation damage is the hypothalamus rather than the pituitary, although the latter may be affected directly. PMID- 12566715 TI - Impact of cancer therapy on the reproductive axis. AB - Cancer therapy includes surgery, chemotherapy and irradiation. Depending on the diagnosis, the location of the neoplasm and the age of the patient, these treatment modalities may be given alone or in combination. All forms of cancer therapy can affect the hypothalamic-pituitary-gonadal axis. The long-term consequences for reproductive function depend on several aspects. The sex of the patient is important, since ovarian and testicular function differ significantly. Sex hormone production in the female is dependent on the presence of germ cells, whereas this is not the case in the male. The sensitivity of germ cells to cancer therapy also differs between the sexes. Moreover, the sensitivity of both the hypothalamic-pituitary axis and the gonads is highly age dependent. With regard to chemotherapy, the possible damage to the gonads is dependent on the total dose and type of agent given. According to current knowledge, the hypothalamic pituitary axis is not affected by conventional doses of chemotherapy. Radiotherapy has by far the most damaging effect on the reproductive axis, having serious adverse effects on both the hypothalamic-pituitary area as well as on the gonads themselves. The harmful effect of irradiation depends on the total dose of irradiation, the radiation field, as well as the number and size of fractions given. The long-term consequences of recently introduced radiotherapeutic methods such as stereotactic irradiation are not yet known. The present review will focus on the late effects of cancer therapy in children and young adults with acute lymphoblastic leukaemia, non-Hodgkin's lymphoma, brain tumour, Hodgkin's lymphoma or Wilms' tumour, including the adverse effects of bone marrow transplantation. PMID- 12566716 TI - Restoration of fertility after treatment for cancer. AB - The late effects of chemotherapy and radiation treatment on fertility are an important issue for long-term survivors of cancer who may not have started or completed a family at the time of diagnosis. Attempts at protecting reproductive function using hormonal manipulation have proved largely unsuccessful and other strategies have to be considered. For men, semen cryopreservation allows subsequent artificial insemination of a female partner or ivf but cryopreserved semen is a finite resource, does not allow natural conception and is not an option for prepubertal boys. In an effort to overcome this, research is in progress to investigate whether testicular cells harvested and cryopreserved before the start of chemotherapy can be reintroduced to the testis after treatment and resume normal spermatogenesis. This has been achieved in a mouse model and the results of experimental protocols in men are awaited with interest. For women, harvested mature oocytes are only poorly tolerant of the freezing process although immediate in vitro fertilization and cryopreservation of embryos can be successful. An experimental technique of great interest is the harvesting and cryopreservation of ovarian cortex before the start of sterilizing treatment. In ewes, the reimplantation of autologous ovarian cortical tissue into surgically castrated animals has resulted in resumption of oestrus, conceptions after normal matings and the birth of live offspring. Recently, ovarian function has been re established using a similar technique in a patient following treatment for Hodgkin's lymphoma, but so far pregnancy has not been reported. PMID- 12566717 TI - Neonatal thyroid disorders. AB - Congenital hypothyroidism is the most prevalent endocrine disorder in the newborn and affects 1 in 3000-4000 newborns. Screening for congenital hypothyroidism is a major achievement of paediatrics because early diagnosis and treatment have resulted in normal development in nearly all cases. The cause of congenital hypothyroidism in the majority of newborns is unknown. However, in some patients the molecular basis of their congenital hypothyroidism has recently been clarified. In patients with congenital hypothyroidism and a normally developed thyroid gland, the autosomal recessive inheritance of loss-of-function mutations of genes encoding for the thyroid peroxidase gene, the sodium-iodide symporter gene and the pendrin gene have been identified. The autosomal recessive inheritance of loss-of-function mutations of the thyroid stimulating hormone (TSH) receptor as well as the dominant inheritance of mutations encoding for transcription factors have been identified in patients with defective thyroid development. Furthermore, it has become evident that in some patients with persistent mental retardation and neurological symptoms, defects of the transcription factor NKX2.1, which is expressed in the thyroid gland as well as in the CNS during embryonic development, cause both defective thyroid and CNS development resulting in persistent neurological and mental defects despite early diagnosis and treatment. Central hypothyroidism is a rare disease with an estimated frequency of not more than 1 in 50000 newborns. Central hypothyroidism can be due to recessive inheritance of loss-of-function mutations of the TSH-beta gene and to developmental defects of the hypothalamus or pituitary. In contrast to the previous assumption that isolated TSH deficiency will not lead to impaired mental development, identification of the molecular defects in central hypothyroidism has clearly demonstrated that some of these patients will have impaired mental development. Clarification of the molecular defects of thyroid development will help to explain the differences in outcome in patients with congenital hypothyroidism and to develop new diagnostic and therapeutic strategies to ensure adequate counselling and care for these patients. PMID- 12566718 TI - The genetics of neonatal hyperinsulinism. AB - Congenital hyperinsulinism (CHI) is the most important cause of persistent hypoglycaemia in the neonate and infant. It is a clinically and genetically heterogeneous entity. The clinical heterogeneity is manifested by severity ranging from extremely severe life-threatening disease to very mild clinical symptoms which may even be difficult to identify. Furthermore, clinical responsiveness to medical and surgical management is extremely variable. Two histopathological forms have been described: a diffuse form of CHI and a focal form of CHI. Recent discoveries have begun to clarify the molecular aetiology of the disease and therefore the mechanisms responsible for its clinical heterogeneity are becoming clearer. Mutations in four different genes have been identified in patients with CHI. Most cases are caused by mutations in genes coding for either of the two subunits of the beta-cell K(ATP) channel (ABCC8 and KCNJ11). In the diffuse form of CHI, the hyperinsulinism is due to a recessive mutation of both alleles of these genes (rare dominant mutations have been described). In the focal form of CHI, two events intervene: first, the inheritance of a paternal ABCC8/KCNJ11 mutation; second, the focal reduction to homozygosity of the mutation during pancreatic development by a localized loss of the maternal 11p15 region. Others cases of CHI are due to rare mutations in the beta-cell enzymes glucokinase (only one family described) and glutamate dehydrogenase in hyperammonaemia-associated hyperinsulinism. However, in as many as 50% of cases, no genetic aetiology has yet been identified. PMID- 12566719 TI - Coronary heart disease: a disorder of growth. AB - A new 'developmental' model for the origins of coronary heart disease and the related disorders of type 2 diabetes, hypertension and stroke is emerging. The finding that people who develop these disorders have altered growth in utero, during infancy and childhood provides a new starting point for research. The immediate prospect for prevention is through protecting infant growth and preventing accelerated weight gain in children made vulnerable to later disease by small size at birth and during infancy. Ultimately we need to optimize maternal diet and composition before and during pregnancy. Despite current levels of nutrition in Western countries the nutrition of many fetuses and infants remains suboptimal because the nutrients available are unbalanced or because their delivery is constrained by the long and vulnerable fetal supply line. PMID- 12566720 TI - Diabetes insipidus. AB - Diabetes insipidus is a heterogeneous condition characterized by polyuria and polydipsia caused by a lack of secretion of vasopressin, its physiological suppression following excessive water intake, or kidney resistance to its action. In many patients, it is caused by the destruction or degeneration of the neurons that originate in the supraoptic and paraventricular nuclei of the hypothalamus. Known causes of these lesions include: germinoma or craniopharyngioma; Langerhans cell histiocytosis and sarcoidosis of the central nervous system; local inflammatory, autoimmune or vascular diseases; trauma following surgery or accident; and, rarely, genetic defects in vasopressin biosynthesis inherited as autosomal dominant or X-linked recessive traits. Thirty to fifty percent of cases are considered idiopathic. Magnetic resonance imaging (MRI) allows identification of the posterior pituitary hyperintensity and of hypothalamic-pituitary abnormalities. Thickening of the pituitary stalk is the second most common finding on MRI scans in several local inflammatory pathologies and autoimmune diseases or germinoma, but it is not specific to any single subtype. A progressive increase in the size of the anterior pituitary gland should alert physicians to the possibility that a germinoma is present, whereas a decrease can suggest the presence of an inflammatory or autoimmune process. Most children with acquired central diabetes insipidus and a thickened pituitary stalk have anterior pituitary hormone deficiencies during follow-up. Biopsy of enlarged pituitary stalk should be reserved for patients with a hypothalamic-pituitary mass and progressive thickening of the pituitary stalk, since spontaneous recovery may occur. PMID- 12566721 TI - Mineralocorticoid mechanisms. PMID- 12566722 TI - Clinical and genetic aspects of phaeochromocytoma. AB - Phaeochromocytoma is a tumour of the adrenal medulla, which, although rare, is a major cause of correctable hypertension with a prevalence of 0.1-0.5% in the hypertensive population. Clinical symptoms include attacks of paroxysmal headache, sweating, palpitations, stress and a sense of imminent death. Often associated with the above is an increase in blood pressure. Despite the fact that the underlying genetic mechanisms of phaeochromocytoma have been well investigated, they are still incompletely understood. In approximately 80% of cases the tumour occurs sporadically, but it may occur in association with type 2 multiple endocrine neoplasia, type 1 neurofibromatosis or von Hippel-Lindau disease. Molecular evidence suggests that other genes such as SDHD or SDHB may control its development; the possibility of other putative phaeochromocytoma genes is currently being investigated. PMID- 12566723 TI - The role of growth hormone in the regulation of protein metabolism with particular reference to conditions of fasting. AB - Growth hormone (GH) has potent protein anabolic actions, as evidenced by a significant decrease in lean body mass and muscle mass in chronic GH deficiency, and vice versa in patients with acromegaly. Depending on the prevailing physiological conditions and on which tissues and which proteins are under examination, the mechanisms involved include both stimulation of protein synthesis and restriction of protein breakdown. Apart from the possible direct effects of GH on protein dynamics, a number of additional anabolic agents, such as insulin, insulin-like growth factor-I and free fatty acids (FFA), are activated. Some of the most recent studies in the field have demonstrated a decisive role of stimulation of lipolysis and high circulating levels of FFA in orchestrating the maintenance of the protein pool of the body. PMID- 12566724 TI - Type 2 diabetes in childhood: the American perspective. AB - The incidence of type 2 diabetes mellitus is steadily escalating throughout the world in people from a wide range of ethnic groups and all social and economic levels. Type 2 diabetes is no longer a disease only of adults: parallel with the global epidemic of type 2 diabetes in adults, an 'emerging epidemic' of type 2 diabetes has been observed in youth over the last decade. Research and clinical experience in adults have established that insulin resistance is a major risk factor for type 2 diabetes. However, insulin resistance alone is not sufficient to cause diabetes, which will develop only when insulin secretion by the beta cells fails. This review discusses the recent emergence of type 2 diabetes in children and adolescents, its risk factors, pathophysiologic mechanisms and treatment modalities. PMID- 12566725 TI - Type 2 diabetes mellitus in children and adolescents: a review from a European perspective. AB - Changes in food consumption and exercise are fueling a worldwide increase in obesity in children and adolescents. As a consequence of this dramatic development, an increasing rate of type 2 diabetes mellitus has been recorded in children and adolescents in the USA and, more recently, in many countries around the world. Both genetic and environmental factors contribute to the pathogenesis of type 2 diabetes. Lower susceptibility in white Caucasians and higher susceptibility in Asians, Hispanics and blacks have been noted. There is a high hidden prevalence and a lack of exact data on the epidemiology of the disease in Europe: in Germany only 70 patients below the age of 15 years were identified in the systematic, nationwide DPV (Diabetessoftware fur prospektive Verlaufsdokumentation) diabetes survey, but our calculations suggest that more than 5000 young people in Germany at present would meet the diagnostic criteria of type 2 diabetes. In Australasia, the prevalence of type 2 diabetes is reportedly high in some ethnic groups and again is linked very closely to the obesity epidemic. No uniform and evidence-based treatment strategy is available: many groups use metformin, exercise programmes and nutritional education as a comprehensive approach to treat type 2 diabetes in childhood and adolescence. The lack of clear epidemiological data and a strong need for accepted treatment strategies point to the key role of preventive programmes. Prevention of obesity will help to counteract the emerging worldwide epidemic of type 2 diabetes in youth. Preventive programmes should focus on exercise training and reducing sedentary behaviour such as television viewing, encouraging healthy nutrition and supporting general education programmes since shorter school education is clearly associated with higher rates of obesity and hence the susceptibility of an individual to acquire type 2 diabetes. PMID- 12566726 TI - Nuclear receptors and co-regulators in adrenal tumors. AB - We have reported that excessive steroid hormone production in adrenal cortical tumors results from the disordered expression and activity of specific steroidogenic enzymes. Since no genetic mutations in these steroidogenic enzymes have as yet been identified, disordered expression at the transcription level may be crucial for excessive hormone production in adrenocortical tumors. Nuclear receptors SF-1 and COUP-TF/DAX-1 have been shown to activate and repress, respectively, the transcription of CYP17 gene in a mutually exclusive manner in Y 1 cells. Interestingly, the expression of COUP-TF and DAX-1 is significantly decreased in the cortisol-producing adenomas, in which CYP17 is overexpressed. Conversely, DAX-1 is highly expressed in deoxycorticosterone-producing adenomas, where CYP17 expression is almost absent. These expression profiles indicate the possibility that COUP-TF and DAX-1 play important roles in the transcriptional repression of CYP17 in adrenal tumors. To clarify the mechanisms of COUP-TF mediated repression, we therefore screened for COUP-TF-interacting proteins using a yeast two-hybrid system from a cortisol-producing adenoma cDNA library. We then cloned a novel COUP-TF-interacting protein-1 (CIP-1), which interacts with COUP TFI, COUP-TFII, and SF-1. Functionally, CIP-1 can act as a transcriptional co repressor for COUP-TF repression activity. CIP-1 expression profiles parallel those of COUP-TFI in steroidogenic tissues, strongly suggesting that, together, COUP-TFI and CIP-1 play an important role in steroidogenesis. PMID- 12566727 TI - SF1 in the development of the adrenal gland and gonads. AB - SF1 (steroidogenic factor-1; NR5A1) is an orphan nuclear receptor that is expressed in the adrenal gland, gonads, spleen, ventromedial hypothalamus and pituitary gonadotroph cells. Combined approaches of targeted mutagenesis in mice and examination of the effects of naturally occurring mutations in humans have clarified the role of SF1 in steroidogenesis and development. Targeted disruption of SF1 (FTZF1) in mice prevents gonadal and adrenal development and causes male to-female sex reversal. A heterozygous loss-of-function human SF1 mutation (G35E) was described in a patient with adrenal failure and complete 46,XY sex reversal, indicating that haploinsufficiency of this transcription factor is sufficient to cause a severe clinical phenotype. In an infant with a similar clinical phenotype, a homozygous SF1 mutation (R92Q) was identified. In functional assays, this mutant SF1 protein exhibited partial loss of DNA binding and transcriptional activity when compared with the more severe G35E P-box mutant. These patients reveal the exquisite sensitivity of SF1-dependent developmental pathways to gene dosage and function in humans. PMID- 12566728 TI - Low birth weight: effect on insulin sensitivity and lipid metabolism. AB - The metabolic and cardiovascular complications associated with reduced fetal growth have been identified during the past 10 years. These complications that encompass cardiovascular diseases and insulin resistance syndrome consist of dyslipidemia, impaired glucose tolerance or type 2 diabetes and appear to result from the initial development of insulin resistance. The association of reduced fetal growth with the other parameters of the syndrome X appear less constant than with insulin resistance and the expression and/or the age of onset seem to depend on the degree of genetic predisposition of the population. Although the mechanisms underlying the development of the insulin resistance associated with reduced fetal growth remain unclear, some evidence argues in favor of a key role of the adipose tissue. Several hypotheses have been proposed over the past 10 years to understand this unexpected association. Each of them points to either a detrimental fetal environment or genetic susceptibilities or interactions between these two components as playing a critical role in this context. Although not confirmed, the hypothesis suggesting that this association could be the consequence of genetic/environmental interactions remains at the moment the most attractive. PMID- 12566729 TI - Growth hormone (GH) secretion in patients with childhood-onset GH deficiency: retesting after one year of therapy and at final height. AB - BACKGROUND: Recent studies have shown that many patients treated with growth hormone (GH) during childhood because of idiopathic GH deficiency (GHD) are no longer GH deficient when retested after cessation of GH therapy when final height is achieved. These patients are labelled as transient GHD. We hypothesized that normalization of GH secretion in transient GHD could occur earlier during the course of GH treatment, which could allow earlier cessation of GH treatment. METHODS: In a retrospective study, GH secretion was re-evaluated after cessation of GH treatment at final height in 43 patients diagnosed during childhood as idiopathic GHD (10 with multiple pituitary hormonal deficiencies (MPHD) and 33 with isolated GHD (IsGHD)). In a prospective study, GH secretion was re-assessed after interruption of GH treatment given for 1 year in 18 children with idiopathic GHD (2 MPHD, 16 IsGHD). GH secretion was evaluated by glucagon or insulin stimulation tests. RESULTS: In the retrospective study, all the 10 patients with MPHD and 64% of the 33 patients with IsGHD were still deficient at re-evaluation using the paediatric criteria to define GHD (GH peak <10 ng/ml at provocative test). The proportion of persisting deficiency was greater in patients with complete IsGHD (86%, 12/14 patients) than in patients with partial IsGHD (47%, 9/19 patients). With the criteria proposed in adulthood (GH peak <3 ng/ml), all the 10 patients with MPHD were still considered to be deficient. In contrast, only 15% (5/33 patients) with IsGHD had a maximal GH value <3 ng/ml (36% of the 14 patients with complete IsGHD and none of the 19 patients with partial IsGHD). In the prospective study, after interruption of GH therapy given for 1 year, the 2 patients with MPHD were still GHD at re-evaluation and they resumed GH treatment. Among the 16 patients with IsGHD, 13 (81%) were still deficient (peak response <10 ng/ml) after 1 year. Two of the 3 patients in whom GHD was not confirmed at retesting after 1 year GH showed again a deficient response at second retesting. CONCLUSIONS: Although many patients diagnosed with IsGHD during childhood have a normalized GH secretory capacity when retested during adulthood, early retesting after interruption of GH treatment given for 1 year during childhood does not enable to determine if GH therapy has to be discontinued before cessation of growth. PMID- 12566730 TI - Effects of hypoxia and thyroid hormone on mRNA levels and activity of phosphoglycerate mutase in rabbit tissues. AB - AIM: In the present work, we studied the effects of hypoxia and triiodothyronine (T(3)) on phosphoglycerate mutase (PGAM) activity and expression in rabbit liver, brain, and skeletal muscle under in vivo conditions. METHODS: Hypoxia was induced in a methacrylate cage with a mixture of 90% nitrogen and 10% oxygen. Hyperthyroidism was induced daily by T(3) injection (250 microg/kg). RESULTS: Hypoxia increases the PGAM activity in liver and brain, tissues which possess type PGAM-BB isozyme, but does not affect the PGAM activity in muscle which possesses type PGAM-MM isozyme. T(3) administration increases the PGAM activity in muscle and liver, but does not affect the enzyme activity in the brain. In all cases, the activity changes in parallel with those of PGAM mRNA levels. CONCLUSION: The tissue-specific effects of hypoxia and T(3) could be explained by the tissue-specific distribution of both PGAM isozyme and T(3) receptors. PMID- 12566731 TI - Pattern of bone markers during pregnancy and their changes after delivery. AB - OBJECTIVE: To investigate whether bone resorption markers change during pregnancy and lactation, and how they are correlated with human placental lactogen (hPL) and PRL. SUBJECTS: Young women before pregnancy, during pregnancy and during a 12 month post-delivery period (study group; n = 22); and age- and weight-matched normal cycling women (control group; n = 22) for a 20-month-period participated in the study. RESULTS: In the study group, women both during pregnancy (from the 8th up to the 38th week) and during a 6-month period of lactation, pyridinoline and deoxypyridinoline urinary levels were significantly higher than those of pre pregnancy and control women. They returned to basal values at the 12th post delivery month. During pregnancy there were early and late peak increases, at the 8th and 32nd week, respectively. At the 32nd, 34th, 36th and 38th week of pregnancy, pyridinoline and deoxypyridinoline urinary values were significantly correlated with hPL serum levels. CONCLUSIONS: During pregnancy the maternal bone resorption seems to vary critically at early and late stages. A complete reversal of these variations seems to occur after lactation. Further studies could evaluate if changes in placental function are capable of differently interfering with maternal bone resorption. PMID- 12566732 TI - Relationship between adrenomedullin and vasopressin-aquaporin system under general anesthesia. AB - AIM: The roles of adrenomedullin (AM) in body fluid balance under general anesthesia were investigated. METHODS: Time course changes in plasma osmolality, AM, arginine vasopressin (AVP), and urinary aquaporin 2 (AQP2) in 17 patients undergoing abdominal surgery under general anesthesia were examined. RESULTS: Increases in plasma AM levels were observed in parallel with increases in the levels of urinary AQP2/creatinine (Cr) before induction and 90 and 180 min after initiation of anesthesia. Significant correlations between plasma AM and urinary AQP2/Cr (r = 0.62, p < 0.0001) as well as urinary AVP/Cr and AQP2/Cr (r = 0.60, p < 0.0001) were uncovered. Multivariate stepwise analysis identified plasma AM as the critical independent factor affecting urinary AQP2/Cr level. CONCLUSION: A novel correlation of AM and AQP2 which overlays an AVP-AQP2 system may play a key role in fluid homeostasis during general anesthesia. PMID- 12566733 TI - Effect of x-irradiation on growth and the expression of parathyroid hormone related peptide and Indian hedgehog in the tibial growth plate of the rat. AB - AIM: To study the effect of irradiation on the longitudinal growth and the expression of parathyroid hormone-related peptide (PTHrP) and Indian hedgehog (IHh) in tibial growth plates of rats. METHODS: At 3 weeks of age, 30 male rats received a single fraction of irradiation (8 Gy) to their right hind limb, and small groups of animals were sacrificed 1, 2, 3, 5, 7, 10, 15, and 26 weeks after irradiation. Weight and length of both irradiated and nonirradiated tibiae were measured, and sections of the tibiae were stained with HE. PTHrP and IHh were visualized using immunohistochemical techniques. RESULTS: Radiation resulted in persistent growth delay of the irradiated tibiae, with a difference in length of more than 10% between the irradiated and the nonirradiated tibiae 15 weeks or more after irradiation. The growth plate architecture was disturbed, and the expression of both PTHrP and IHh was decreased in the irradiated tibiae. CONCLUSION: As PTHrP and IHh are key regulators of both the pace and the synchronization of the differentiation of growth plate chondrocytes, the reduced expression of PTHrP and IHh may contribute to the changes found after irradiation. PMID- 12566734 TI - Castleman's disease presenting as a goiter in a child. PMID- 12566735 TI - Concomitant association of thyroid sarcoidosis and Graves' disease. AB - OBJECTIVE: Graves' disease (GD) with sarcoid involvement of the thyroid gland has rarely been reported. METHOD: We report a case of GD with thyroid sarcoidosis in a 28-year-old woman. Thyroid function was assessed by triiodothyronine (T(3)), thyroxine (T(4)), thyroid-stimulating hormone (TSH) and TSH receptor antibodies (TSH-R Ab). Thyroid scintigraphy, ultrasound and fine-needle aspiration biopsy were performed. The patient underwent surgery. RESULT: The patient had a nodular goiter. Serum T(3), T(4) and TSH-R Ab levels were elevated with suppressed TSH level. Scintigraphy showed diffuse activity as seen in GD, and ultrasound revealed that parenchyma was heterogenous. Sarcoidosis was discovered on routine chest X-ray. Although no sarcoid involvement was found on specimen, the thyroid gland showed non-caseating granulomas on histology. CONCLUSION: Since sarcoid involvement of the thyroid gland can cause hypofunction, we report the uncommon infiltration of sarcoidosis with hyperthyroidism. PMID- 12566736 TI - Familial form of thyroid dysgenesis: report of thyroid hemiagenesis with accompanying Graves' disease in a woman whose daughter has thyroid agenesis. AB - The subject is a 44-year-old female with thyroid hemiagenesis, who initially presented with hyperthyroidism. Thyroid peroxide antibody and thyroglobulin antibody levels were high. The scintiscan study and sonographic findings were compatible with thyroid hemiagenesis with accompanying Graves' disease. In reviewing her family history, her daughter was found to have thyroid agenesis, and other thyroid disorders were found in 2 female family members. PMID- 12566737 TI - A case of Turner syndrome with congenital hypothyroidism untreated until age 38 years. AB - OBJECTIVE AND METHODS: The effect of thyroid hormone on human growth and maturation is considered 'permissive'. To evaluate the effect of a prolonged thyroid hormone defect, especially in the pubertal period, a woman with untreated congenital hypothyroidism underwent studies of thyroid function and bone maturation for the first time at age 38 years 10 months and received thyroid hormone replacement. RESULTS: The karyotype was 45,X/46,XX. Menstruation had occurred for 10 years, from menarche until she was about 31 years old. Epiphyseal closure of the left hand was incomplete. The serum thyroid hormone level was virtually undetectable, and her thyroid gland was not detectable in the normal position by ultrasonography. Her height increased by 3.5 cm in the first 9 months after starting thyroid hormone replacement; after 11 months, closure of the epiphysis was complete. CONCLUSION: Thyroid hormone is necessary to achieve bone maturation and epiphyseal closure, but its role is only permissive. PMID- 12566738 TI - Efficient score statistics for mapping quantitative trait loci with extended pedigrees. AB - The method of variance components is the method of choice for mapping quantitative trait loci (QTLs) with general pedigrees. Being a likelihood-based method, this method can be computation intensive even for nuclear families, and has excessive false positive rates under some situations. Here two efficient score statistics to detect QTLs are derived, one assumes that the candidate locus has no dominance effect, and the other one does not make such an assumption. These two score statistics are asymptotically equivalent to the method of variance components but they are easier to compute and more robust than the likelihood ratio statistic. The derivation of these score statistics is facilitated by separating the segregation parameters, the parameters that describe the distribution of the phenotypic value in the population, from the linkage parameters, the parameters that measure the effect of the candidate locus on the phenotypic value. Such a separation of the model parameters greatly reduces the number of parameters to be dealt with in the analysis of linkage. The asymptotic distributions of both score statistics are derived. Simulation studies indicate that, compared to the method of variance components, both score statistics have comparable or higher power, and their false-positive rates are closer to their respective nominal significance levels. PMID- 12566739 TI - Ancestral founder of mutation W283X in the porphobilinogen deaminase gene among acute intermittent porphyria patients. AB - Acute intermittent porphyria (AIP) is a low-penetrant autosomal dominant disorder caused by mutations in the porphobilinogen deaminase gene (PBGD). Nearly 60% of all Swiss AIP patients carry a nonsense mutation W283X (G(7916)-->A). In France, the prevalence of W283X is <5%. To determine whether W283X was a founder mutation or originated from multiple de novo events, we studied 25 apparently unrelated W283X families and index patients, 21 of Swiss and 4 of French origins. In the absence of sufficient genealogical data to verify the ancestral background of these W283X families/patients, we identified haplotypes of seven intragenic single nucleotide polymorphisms (SNPs) in the PBGD gene as well as eight microsatellites flanking the PBGD gene covering 9.88 cM in chromosome 11. Molecular cloning and sequencing experiments were required in order to completely resolve the intragenic haplotypes in this study cohort which mainly consisted of single index patients and families with limited members. Thirteen of the 25 W283X families/patients carry a SNP haplotype [C-A-A-A-G-C-W283X-G] and 12 (including four French families) carry a [T-G-G-G-G-C-W283X-G] haplotype. A less conserved microsatellite haplotype was identified among the 25 W283X alleles which allowed us to estimate the age of the mutation. Since W283X is not explained by a methylcytosine mutation, we favor the hypothesis of a single mutational event which took place on the [T-G-G-G-G-C-G] background at approximately 40 generations or 1000 years ago. Around 550 years ago, a recombination event occurred between intron 3 and 10 of the PBGD gene which resulted in the [C-A-A-A G-C-W283X-G] haplotype only found in a restricted region. PMID- 12566740 TI - Two new recursive likelihood calculation methods for genetic analysis. AB - Recursive likelihood calculations for genetic analysis with ungenotyped pedigree data employ variations of the Elston-Stewart (ES) or the Lander-Green (LG) algorithms. With the ES algorithm, the number of loci may be limited but not the pedigree size. With the LG algorithm, the reverse is the case. We introduce two new algorithms for the computation of regressive likelihoods for pedigrees with multivariate traits. The first is an alternative formulation of our existing model, which leads to a simpler form in the binary trait, polygenic and mixed model cases. The second is an approximation model, which is computationally efficient. These methods apply to both continuous and binary traits, in the oligogenic and polygenic cases. Both methods coincide in the binary case. We considered these methods for cases in which all the traits are controlled by a single locus, with each trait controlled by one locus independent to the others. Simulation studies and analysis of a real data are presented for segregation analysis as illustrations. These methods can also be used in other model-based analyses. These methods are implemented in G.E.M.S., the genetic epidemiology models software. PMID- 12566741 TI - Enhanced pedigree error detection. AB - Accurate information on the relationships among individuals in a study is critical for valid linkage analysis. We extend the MLLR, EIBD, AIBS and IBS tests for detection of misspecified relationships to a broader range of relative pairs, and we improve the two-stage screening procedure for analyzing large data sets. We have developed software, PREST, which calculates the test statistics and performs the corresponding hypothesis tests for relationship misclassification in general outbred pedigrees. When a potential pedigree error is detected, our companion program, ALTERTEST, can be used to determine which relationships are compatible with the genotype data. Both programs are now freely available on the web. PMID- 12566742 TI - Endometriosis, tampons and orgasm during menstruation: science, press and patient organizations. PMID- 12566743 TI - Concerning the article by Meaddough et al: Sexual activity, orgasm and tampon use are associated with a decreased risk for endometriosis. PMID- 12566744 TI - Concerning the article by Meaddough et al: Sexual activity, orgasm and tampon use are associated with a decreased risk for endometriosis. PMID- 12566745 TI - Arcus tendineus levator ani tear causing pelvic hematoma after tension-free vaginal tape insertion. AB - Although vaginal tamponade is usually sufficient for hemostasis, it is not useful if a pelvic hematoma occurs. Here, we report the unusual case of a large pelvic hematoma necessitating laparotomy for hemostasis due to a tear of the arcus tendineus levator ani after insertion of a tension-free vaginal tape. PMID- 12566746 TI - Injection therapy for the treatment of stress urinary incontinence in women. AB - The purpose of this review is to summarize the results of reports on injectable agents in the treatment of female stress urinary incontinence. Six agents were reviewed: Teflon; autologous fat; collagen; silicone microparticles; silicone microballoons, and pyrolytic carbon. Collagen was the most frequently reported agent and yielded a 1-year cure/improvement rate of 60-80%, but results worsened significantly with longer follow-up. Teflon has been used longer in the treatment of stress incontinence, but low long- and short-term success rates and reported complications such as particle migration have prevented its widespread acceptance. Autologous fat has been suggested as a natural injectable but its success rate has been disappointing. The injection of silicone microparticles is associated with a long-term success rate of about 70% in patients with intrinsic sphincter deficiency, and it can now be injected more easily without the need of urethroscopy. Silicone microballoons and pyrolytic carbon has recently been introduced into clinical practice with a short-term success rate of about 70%, but longer follow- up is needed. In conclusion, the long-term durability, cost effectiveness, and some safety issues still have to be addressed by further clinical trials. PMID- 12566747 TI - Differences in umbilical cord blood units collected during cesarean section, before or after the delivery of the placenta. AB - Umbilical cord blood is largely employed as an alternative source of stem cells in the treatment of hemato-oncological diseases. Current results show that the success rate of purified umbilical cord blood engraftment is comparable to that obtained using bone marrow, and it is directly related to the number of pluripotent stem cells transplanted. The technique of fetal blood collection varies among different umbilical cord blood banks. Many authors collect umbilical cord blood during vaginal delivery, after placental detachment, while others collect it while the placenta is still within the uterus. In a previous randomized trial, we showed a greater collection of umbilical cord blood before placental detachment during vaginal delivery. The present study was performed to determine whether umbilical cord blood collection before placental detachment (group A) during cesarean section is superior to that after placental delivery (group B) puncturing the umbilical vein once and using a closed bag system. To accomplish this, 47 pregnant women subjected to cesarean section were enrolled in the study. Twenty-one of them were allocated to group A, while the remaining 26 formed group B. The volume of umbilical cord blood collected from the patients of group A was greater than that collected from patients of group B. The cord blood volume collected was 90.7 +/- 6.0 versus 60.9 +/- 13.7 ml; the cord blood nucleated cell number was 10.1 +/- 1.2 x 10(8) vs. 7.1 +/- 0.8 x 10(8); and the mean cord blood CD34+ cell number was 20.0 +/- 6.0 x 10(5) vs. 16.4 +/- 2.4 x 10(5), respectively. PMID- 12566748 TI - Conservative management of cervical intraepithelial neoplasia (CIN(2-3)) in pregnant women. AB - The management of cervical intraepithelial neoplasia (CIN(2-3)) diagnosed during pregnancy was the subject of this study. Two hundred and eight pregnant women with an abnormal cytology were assessed in our unit over a 10-year period. The age of the patients ranged from 20 to 45 (mean 28) years. Seventy-eight of these women were histologically proven to have CIN(2-3). All patients were followed up every 8-10 weeks by cytology and colposcopy during pregnancy and reassessed 8-12 weeks postpartum. The disease persisted in 30 cases (38.4%), whereas in the remaining 48 cases it regressed to CIN(1). No case of invasive disease developed during the follow-up period in these pregnant patients. Conservative management of CIN(2-3) during pregnancy is acceptable, but close follow-up and colposcopic expertise are necessary. PMID- 12566749 TI - Behaviour of cytokine levels in serum and peritoneal fluid of women with endometriosis. AB - Endometriosis is a disorder characterised by presence and growth of endometrial tissue outside the uterus, primarily into the peritoneum. The peritoneal fluid (PF) of women with endometriosis undergoes a number of biological changes, including local inflammatory-reparative phenomena and peripheral blood mononuclear cells (PBMC) involvement. These activated cells as well as the endometriotic cells secrete various cytokines with pleiotropic biological activities. Dynamic interplay among cytokines may contribute to realise a favourable microenvironment for the implantation of endometrial cells and the progression of the disease. In the present study, we evaluated the levels of cytokines, such as the tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) in PF and in serum (S) of women with endometriosis to compare their behaviour in both biological fluids. The patients (n = 26) were women of reproductive age attending our observation centre for infertility, diagnosed endometriosis at laparoscopy. Control group (n = 5) consisted of women affected by non-immunologic infertility, diagnosed by explorative laparoscopy. S samples were obtained from peripheral blood before anaesthesia and laparoscopy. PF samples were collected at the time of laparoscopy. Both biological fluids were examined for cytokine by ELISA assays. Our results showed that S and PF levels of TNF-alpha, not dosable in controls, were very high at the early stage and decreased significantly with the severity of the disease (p < 0.001). TGF-beta levels were significantly (p < 0.001) higher than in controls and increased with the severity of the disease (p < 0.001), particularly in the PF. S and PF IL-8 as well as MCP-1 concentrations at all stages were higher than in controls (p < 0.001), yet showed an opposite behaviour in both biological fluids. In fact, S levels of IL-8 and MCP-1 were significantly (p < 0.001) higher at early stages and decreased with the severity of the disease, whereas we observed a significant (p < 0.001) enhancement of these chemokine levels in PF from stage I to stage II and stage III. These observations showed that TNF-alpha and TGF-beta levels were overlapping in S and PF of women with endometriosis. On the contrary, MCP-1 and IL-8 S concentrations decreased with the severity of the disease, whereas PF levels showed markedly increased at severe stages. Taken together the observed changes may be due both to the increased peritoneal macrophage activity and to the larger recruitment of PBMC and autocrine release by endometriotic cells. PMID- 12566750 TI - Postmenopausal hormone replacement therapy use decreases oxidative protein damage. AB - OBJECTIVES: The aim of this study is to evaluate oxidative protein damage (OPD) by investigating protein carbonyl (PCO) and nitrotyrosine (NT) levels, oxidative stress by total thiol (T-SH), erythrocyte glutathione (GSH) and nitric oxide (NO) levels in women receiving hormone replacement therapy (HRT). MATERIALS AND METHODS: To examine the influence of oxidative stress on OPD, we studied 12 postmenopausal women who had received HRT for 6 months, and 13 postmenopausal women who did not receive HRT, as the control group. All subjects were non smokers. Blood samples were drawn in the fasting state and processed within 1 h of collection. For NT and NO, serum samples were stored at -70 degrees C until analysis; all other parameters were determined on the same day of collection. RESULTS: After 6 months, plasma PCO and T-SH levels were decreased, GSH and NO levels were increased, and NT levels were not changed in 12 postmenopausal women receiving HRT. Except the NT levels, the rest of the parameters did not significantly change in the control group. Interestingly, mean NT levels in the control group increased significantly. CONCLUSIONS: A crucial part of the protective effect of HRT on the cardiovascular system arises secondary to the interaction between estrogen and vessel wall. Our results suggest that an important component of the mechanism underlying this interaction may depend on estrogen's antioxidant effect and its preventive role in OPD. PMID- 12566751 TI - Impact of insulin resistance on pregnancy complications and outcome in women with polycystic ovary syndrome. AB - The aim of the study was to determine the risk of developing gestational diabetes mellitus (GDM) and pregnancy-induced hypertension (PIH)/pre-eclampsia in a cohort of pregnant women with the polycystic ovary syndrome (PCOS) and known insulin sensitivity status. Pregnancies and neonatal outcome were recorded in a prospective cohort study comprising 29 non-insulin-resistant PCOS women, 23 insulin-resistant PCOS women and a control group of 355 women who had conceived after assisted reproduction. Hypertension, pre-eclampsia and GDM were recorded as well as pregnancy duration, method of delivery and birth weight. The frequency of hypertension was significantly elevated in PCOS women (11.5%) compared to controls (0.3%), p < 0.01. However, the frequency of pre-eclampsia was significantly elevated only in the insulin resistant PCOS women (13.5%) compared to controls (7.0%), p < 0.02. GDM was significantly more frequent in PCOS women (7.7%) than controls (0.6%), p < 0.01. Insulin resistance prior to pregnancy, determined by continuous infusion of glucose with model assessment (CIGMA) test, did not further increase the frequency of GDM. Newborns from PCOS pregnancies were significantly more often delivered by Caesarean section than controls (40.3 vs. 27.3%, p < 0.05) and transferred to neonatal intensive care unit more often than controls (19.2 vs. 9.0%, p < 0.01). Thus we show that the frequencies of pre eclampsia and GDM are increased in PCOS pregnancies. PMID- 12566752 TI - Adrenomedullin concentrations in early 2nd-trimester amniotic fluid: relation to preterm delivery and fetal growth at birth. AB - OBJECTIVE: The purpose of this study was to evaluate whether adrenomedullin concentrations in the early 2nd-trimester amniotic fluid predict preterm delivery or fetal growth at birth. METHODS: The adrenomedullin concentrations in early 2nd trimester amniotic fluid were measured in 70 pregnancies with term delivery and in 3 pregnancies with preterm delivery. Total and free adrenomedullin concentrations were measured from early 2nd-trimester amniotic fluid samples using an immunoradiometric assay. RESULTS: The amniotic fluid total adrenomedullin concentrations in women with preterm delivery were significantly higher (129.7 +/- 19.6 fmol/ml) than those in women with term delivery (92.5 +/- 28.2 fmol/ml; p < 0.05). There were no significant differences for amniotic fluid free adrenomedullin concentrations and free/total adrenomedullin ratios between the two groups. Total or free adrenomedullin concentrations in the early 2nd trimester amniotic fluid showed an inverse correlation both with birth weight (r = 0.27, p < 0.05, and r = 0.21, p < 0.05) and height (r = 0.30, p < 0.05, and r = 0.28, p < 0.05). There were no correlations between placental weight and total or free adrenomedullin concentrations in the early 2nd-trimester amniotic fluid. CONCLUSIONS: These results suggest that adrenomedullin concentrations in the early 2nd-trimester amniotic fluid might be related to further in utero fetal growth and that high levels of adrenomedullin in the early 2nd-trimester amniotic fluid may be involved in the occurrence of preterm delivery. PMID- 12566753 TI - Comparison of the clinical efficacy and safety of flutamide versus flutamide plus an oral contraceptive in the treatment of hirsutism. AB - OBJECTIVE: To compare the clinical, hormonal, biochemical and hemotological effects of flutamide versus flutamide plus an oral contraceptive (OC) in the treatment of hirsutism. PATIENTS: Eighty-four women with hirsutism were equally randomized to receive either flutamide or flutamide plus OC. DESIGN: Eighty-four women with hirsutism were recruited from patients presenting to our hospital. Each patient underwent a complete medical and gynecological examination as well as blood cell counts, biochemical and endocrine profiles. Hirsutism scores and laboratory tests were done during the 1st, 3rd and 6th months of therapy. Thirty seven women in the flutamide group (taking 250 mg flutamide per day) and 32 women in the flutamide plus OC group (taking 250 mg flutamide plus 35 microg ethinyl estradiol and 2 mg cyproterone acetate per day) regularly followed the therapy regimens. RESULTS: There were no significant differences in Ferriman-Gallway scores at the beginning and at the end of the therapies between the two groups (p > 0.05). At the 6th month, the decreases in Ferriman-Gallway scores were significant in both flutamide (from 18.95 +/- 4.44 to 14.46 +/- 5.02; p < 0.05) and flutamide plus OC groups (from 19.94 +/- 4.31 to 15.58 +/- 4.28; p < 0.05). In the first group, 2 of 6 oligomenorrheic women had regular cycles at the end of the therapy. Oligomenorrhea in 8 women was all changed to regular cycles in the flutamide plus OC group. Regarding the hormonal profile, only in the second group were prolactin, free testosterone and dehydroepiandrosterone sulfate levels significantly decreased (p < 0.05). Hepatic function tests were significantly increased in both groups, but they were all within normal ranges. CONCLUSION: These data suggest that both therapies were similarly effective and safe in the treatment of hirsutism. In women with oligomenorrhea and/or needing contraception, adding an OC shows better results than flutamide treatment alone in providing regular cycles. PMID- 12566754 TI - Dysgerminoma in a patient with a tumor of the neck. Empiric treatment of stage IV dysgerminoma. AB - The evolution of therapy for malignant ovarian germ cell tumors is one of the true success stories in oncology. Treatment outcome has improved greatly thanks to cisplatin-based combination chemotherapy. According to the well-established treatment guidelines for advanced cases, we treated a case of stage IV undifferentiated germ cell tumor in which we were able to preserve the patient's fertility. We concluded that the PEP regimen is an effective treatment for the patient with metastatic germ cell tumor. PMID- 12566755 TI - Circulating inhibin forms in patients with hydatidiform mole. AB - Circulating inhibin forms have not been evaluated in patients with hydatidiform mole (HM) before evacuation of the mole. Serum inhibin A, inhibin B, pro-alphaC and hCG levels were determined by enzyme-linked immunosorbent assay in HM before and after evacuation of the mole (inhibin B only before evacuation). Before evacuation, inhibin B levels in HM were undetectable, and inhibin A levels were similar to those in normal women at the corresponding weeks of pregnancy. Pro alphaC and hCG levels were higher in molar pregnancy than in normal pregnancy. After evacuation, pro-alphaC levels significantly fell in the patients with spontaneous resolution, but not in those who developed persistent trophoblastic disease (PTD). Both pro-alphaC and hCG levels were higher in the patients with PTD than in those with spontaneous resolution after evacuation, reflecting continued stimulation of the corpus luteum by lower, but still effective hCG levels, persistent trophoblastic tissues secreting pro-alphaC or both. PMID- 12566756 TI - Ultrasound assessment of biometric trends in a case of thanatophoric dysplasia. AB - We present a case of thanatophoric dysplasia diagnosed at the 21st week of gestation. Serial ultrasound was performed throughout pregnancy. The scans showed a distinctive pattern of development of the fetal long bones. Up to week 25, the fetal long bones appeared to grow steadily but slower compared to normal measurements (4-5 SD below the mean); then, between weeks 26 and 30, long bone growth was further and more severely hampered, until it almost stopped altogether approaching term, with measurements 9-12 SD below the mean at week 38. PMID- 12566757 TI - Diet and feeding behaviour of Indri indri in a low-altitude rain forest. AB - The diet and feeding behaviour of Indri indri were investigated in the Betampona Reserve, eastern Madagascar, over 12 months from February 2000 to February 2001. The highly folivorous diet of this species was confirmed--feeding on foliage (leaves and petioles) accounting for 82% of feeding records. Immature leaves were the preferred dietary item, but at times of relative scarcity mature leaves, fruit, seeds, flowers and bark were substituted. The indri were observed to feed on parts from 22 plant families, 37 genera and at least 42 species. The most important plant families in the diet of Indri were Lauraceae, Clusiaceae and Myristicaceae. Most feeding at Betampona was observed at 5-20 m above the forest floor amongst small (2.1-5.0 cm), oblique/horizontal supports (0-45 degrees). The indri spent 41.4% of their active period feeding most commonly in above-branch postures. Studies such as this are important for the development of conservation management plans for this endangered species. PMID- 12566758 TI - Mitochondrial COII gene sequences provide new insights into the phylogeny of marmoset species groups (Callitrichidae, Primates). AB - Mitochondrial cytochrome oxidase II (COII) gene sequences (549 base pairs) were used to investigate the taxonomic relationships among 12 marmoset (Callithrix, Cebuella and Mico) taxa. A large number of substitutions were found in the third base codon positions, providing a strong phylogenetic signal in a gene coding a conserved protein. Despite the significant affinity between the 2 Amazonian genera Cebuella and Mico, found in recent molecular studies, the analysis presented here did not resolve convincingly the phylogenetic relationships between the 3 genera. Mico nevertheless formed 3 distinct clades, reflecting a basic division of species groups based on geographic distribution (east or west of the Rio Tapajos) rather than morphology (presence or absence of auricular hair). This supports the taxonomic distinction of the allopatric emiliae forms. In Callithrix, Callithrix aurita forms a distinct clade, but the remaining morphotypes form a somewhat contradictory cluster, possibly resulting from an extremely rapid radiation. PMID- 12566759 TI - A reappraisal of the locomotion and habitat preference of Theropithecus oswaldi. AB - The one modern member of the genus Theropithecus, T. gelada (Primates, Cercopithecidae), inhabits grassland and is highly terrestrial. It is often supposed that Theropithecus oswaldi, one of the most common primates of the Plio Pleistocene of East and southern Africa, was also a highly terrestrial open habitat species. Ecomorphic analysis was used to assess the locomotor strategy and habitat preference of T. oswaldi, and it was found that this species was unlikely to have had a locomotor strategy and habitat preference identical to that of T. gelada, with T. oswaldi possibly using arboreal substrates in a manner similar to some modern baboon groups. Thus, it appears that there has been considerable ecological diversity in the genus Theropithecus over the course of its evolution, mirroring the diversity evident in the hominin fossil record. PMID- 12566760 TI - Hunting of mammalian prey by Budongo Forest chimpanzees. PMID- 12566761 TI - Lip and tongue function differently affected in individuals with multiple sclerosis. AB - Clinical dysarthria test scores on lip function and tongue function were compared for 77 dysarthric as well as non-dysarthric subjects with multiple sclerosis (MS) and 15 control subjects. Results show that tongue function was significantly more severely affected than lip function in individuals with MS. Furthermore, tongue function, but not lip function, was significantly more severely affected in the MS individuals with no dysarthria compared to the control group. Test items requiring increased rate of movement (oral and verbal diadochokinesis) were significantly more severely affected than the items requiring range and force of movement, but only in the dysarthric MS subgroup. Moderate correlations were found between tongue and lip function and neurological deficit scores, number of years in disease progression, and perceptually perceived consonant and vowel precision. Consequently, tongue dysfunction can be detected clinically and subclinically using a dysarthria test procedure, and as an early sign of articulatory dysfunction it should be an early target in therapeutic interventions. PMID- 12566762 TI - [Suitability of the Munich List of Quality of Life Dimensions (MLDL) for use in logopedic therapy research]. AB - Public health policy increasingly focuses on criteria of cost and cost effectiveness for health care financing. However, tools for assessing the effects of logopedic therapy are largely missing. This study investigated the suitability of the Munich List of Quality-of-Life-Dimensions (MLDL) for use with patients with dysphonia. Twenty-five adults with voice disorders were administered the MLDL at the beginning and at the end of therapy (about 20 sessions). Results were that, although most patients expressed high satisfaction with the therapy received, MLDL scores did not reflect this. Only 2 out of 19 items of the scale indicated a significant improvement in the quality of life after therapy ("ability to relax", p < 0.05, and "satisfaction with the treatment of the disorder", p < 0.01). The MLDL is not sensitive enough to detect all effects of logopedic therapy. The development of specific scales for the detection of effects of logopedic intervention is discussed. PMID- 12566763 TI - Lip kinematics for /p/ and /b/ production during whispered and voiced speech. AB - In the absence of voicing, the discrimination of "voiced" and "voiceless" stop consonants in whispered speech relies on such acoustic cues as burst duration and amplitude, and formant transition characteristics. The articulatory processes that generate these features of whispered speech remain speculative. This preliminary investigation examines the articulatory kinematics differences between whispered /p/ and /b/, which may underlie the acoustic differences previously reported for these sounds. Computerized video-tracking methods were used to evaluate kinematic differences between voiced and voiceless stops. Seven subjects produced the target utterances "my papa puppy" and "my baba puppy" in voiced and whispered speech modes. The results revealed that mean peak opening and closing velocities for /b/ were significantly greater than those for /p/ during whispered speech. No differences in peak velocity for either oral closing or opening were observed during voiced speech. The maximum distance between the lips for oral opening for /b/ was significantly greater than for /p/ during whisper, whereas no difference was observed during voiced speech. These data supported the suggestion that whispered speech and voiced speech rely on distinct motor control processes. PMID- 12566764 TI - Separate detection of vocal fold vibration by optoreflectometry: a study of biphonation on excised porcine larynges. AB - This study describes a novel system allowing the noncontact measurement of vibration of each vocal fold. The system is based on reflectometry, i.e. measurement of light bouncing off a vibrating object. The system works in natural light and is sensitive, compact, and inexpensive. We have used this system for separate and simultaneous detection of the vibration of each vocal fold in excised porcine larynges. Normal conditions of vibration and asymmetry of tension leading to biphonations were studied. Analysis of each vibrating cycle reveals interaction between the vocal folds allowing synchronous vibration despite their physical asymmetry or episodes of period doubling. PMID- 12566765 TI - Differential modes of alaryngeal communication and long-term voice outcomes following pharyngolaryngectomy and laryngectomy. AB - Patterns of vocal rehabilitation for 37 pharyngolaryngectomy patients and 55 total laryngectomy patients over a 5-year period were compared. An electrolarynx (EL) was introduced as the initial communication mode immediately after surgery for 98% of patients, with 30% of pharyngolaryngectomy and 74% of laryngectomy patients subsequently developing tracheoesophageal speech (TES) as their primary mode of communication. Follow-up with 14 of 37 pharyngolaryngectomy patients and 36 of 55 laryngectomy patients was conducted 1-6 years following surgery and revealed that 90% of the pharyngolaryngectomy patients maintained the use of TES in the long term compared to 69% of the laryngectomy group. Long-term outcomes relating to communication disability and handicap did not differ significantly between the two surgical groups, however the laryngectomy patients had significantly higher levels of wellbeing. Across the whole group of patients, statistical comparison revealed that patients using TES had significantly lower levels of disability, handicap and distress than EL users. Considering that lower levels of disability, handicap and distress are associated with TES, and the data supports that suitably selected patients can maintain functional TES in the long term, increased application of this form of communication rehabilitation should be encouraged where viable for the pharyngolaryngectomy population. PMID- 12566766 TI - Heterotopic pregnancy: successful management with aspiration of cornual/interstitial gestational sac and instillation of small dose of methotrexate. AB - OBJECTIVES: Terminating a corneal pregnancy without jeopardizing a coexistent intrauterine pregnancy. METHODS: A 29-year-old woman with a 4-year history of primary infertility became pregnant after in-vitro fertilization and embryo transfer. A heterotopic pregnancy with a left cornual and intrauterine sacs was confirmed by early transvaginal scan examinations. The ectopic sac was aspirated under ultrasound guidance followed by local injection of 12.5 mg methotrexate. This was followed by serial ultrasound scans and serum betahCG assays. RESULTS: The fetal pole in the ectopic pregnancy disappeared following the procedure. She had no significant vaginal bleeding or any other unusual symptoms. The intrauterine pregnancy progressed satisfactorily until spontaneous onset of labour and vacuum extraction delivery at 39 weeks. The baby weighed 2.9 kg. CONCLUSIONS: Local injection of low-dose methotrexate following aspiration of a cornual pregnancy proved effective in halting the ectopic trophoblasts' growth without adversely affecting a coexistent intrauterine pregnancy. The injected ectopic trophoblasts resolved quickly despite the small dose of methotrexate used. This technique avoided a more costly surgical treatment with its associated risks in subsequent pregnancies. PMID- 12566767 TI - Neural tube defects in the offspring of thalassemia carriers. AB - OBJECTIVES: The risk of neural tube defects (NTDs) is negatively correlated with maternal folate levels in early pregnancy. Thalassemia carriers often have reduced red cell folate contents. The aim of this study was to verify whether thalassemia carriers are at increased risk of having offspring with a NTD. METHODS: We retrospectively evaluated the prevalence of thalassemia carriers among 75 women who had prenatal diagnoses of NTDs made at our center. RESULTS: The prevalence of beta-thalassemia carriers among the affected pregnancies was significantly higher than in the general population (22.5 versus 14%, p < 0.05, chi(2) test). CONCLUSION: Although inconclusive, these data indicate that folate supplementation is mandatory among women with thalassemia who are planning a pregnancy. PMID- 12566768 TI - Umbilical cord pseudocyst in a fetus with trisomy 18. AB - An umbilical cord pseudocyst was detected in the 28th week of gestation in a fetus complicated with growth restriction and polyhydramnios. The combination of cord pseudocysts, growth restriction, and polyhydramnios prompted us to perform a detailed ultrasonographic examination (gray scale and three-dimensional), which revealed the presence of micrognathia, overlapping fingers, and congenital heart defects, features characteristic of trisomy 18. Karyotyping confirmed a diagnosis of trisomy 18. After spontaneous labor onset, the infant was delivered at 31 weeks of gestation, and died soon after delivery. An umbilical cord pseudocyst is a good marker for the prenatal detection of trisomy 18. PMID- 12566769 TI - Fetal biophysical profile and cerebro-umbilical ratio in assessment of perinatal outcome in growth-restricted fetuses. AB - AIM: To examine the value of the fetal biophysical profile (FBP) and the Doppler cerebro-umbilical ratio (C/U) in the assessment of perinatal outcome in growth restricted and hypoxic fetuses. STUDY DESIGN: The prospective clinical study included 87 pregnant women with singleton pregnancies at 28-42 weeks of gestation with clinically verified intrauterine growth restriction (IUGR). After assessment of FBP, flow velocity waveforms from the umbilical and middle cerebral arteries were recorded and the C/U ratio was calculated. The C/U ratio and FBP were assessed twice a week. At delivery, umbilical arterial pH, the occurrence of meconium-stained amniotic fluid (MAF), Apgar score at 5 min and the incidence of cesarean sections were used as outcome parameters. RESULTS: The mean FBP value was 5.5 +/- 0.96 in cases with MAF, and 6.88 +/- 0.26 in cases without MAF. Also, there was no statistical significance in the relation between the C/U ratio and the appearance of MAF. At pH >7.2, the mean FBP value was 7.11 +/- 0.23, while it was 2.83 +/- 0.79 in newborns with acidosis. Both FBP and C/U values were statistically correlated with pH (p < 0.01). The mean umbilical arterial pH was 7.31 +/- 0.0 at a C/U ratio of >/=1 and 7.21 +/- 0.03 at a C/U ratio of <1. In cases with Apgar scores of 8-10, the mean FBP value was 7.28 +/- 0.23, at Apgar scores of 5-7 it was 3.9 +/- 0.52, while at Apgar scores of 0-4 the mean FBP value was 1.5 +/- 0.5. The mean Apgar score at 5 min was 9.54 +/- 0.09 at a C/U ratio of >/=1, and 8.12 +/- 0.49 at a C/U ratio of <1. The mean FBP value in cases of vaginal delivery was 7.55 +/- 0.31. In cesarean section deliveries, the mean FBP value was 5.97 +/- 0.37. Also, there was a high frequency of cesarean sections in growth-restricted fetuses with a C/U ratio of <1 (p < 0.05), i.e. slightly less than FBP. CONCLUSIONS: FBP and C/U ratio were associated with low arterial pH, low Apgar score and the rate of cesarean sections (p < 0.05), but there was no association between FBP or C/U ratio and the appearance of MAF (p > 0.5). Due to their good predictive value the FBP and C/U ratio could be used in the prenatal monitoring of growth-retarded and hypoxic fetuses. These two methods can be used as important parameters in the decision to end pregnancies with IUGR, when pathological values occur. Thus a reduction in perinatal morbidity, mortality and the incidence of infants with poor neurologic outcome can be expected. PMID- 12566770 TI - Pre- and perinatal relations of hemophilia A and B. AB - OBJECTIVE: The authors conducted a retrospective study of obstetric and genetic data, obstetric problems, and pregnancy outcome by investigating 149 pregnancies of patients who received genetic counselling because of hemophilia A or B over a 20-year period. METHODS: In cases with a heterozygous mother, fetal sex was determined. In 23 of 35 cases with male fetuses, a DNA examination was performed. In cases with hemophilic male fetuses, the couple made a decision on whether or not to continue the pregnancy after thorough counselling regarding genetic risk. Hemophilia A occurred 135 pregnancies (98 pregnancies from 55 heterozygous mothers and 37 pregnancies from 20 hemophilic fathers). Hemophilia B occurred in 14 pregnancies (9 pregnancies from 3 heterozygous mothers and 5 pregnancies from 4 hemophilic fathers). RESULTS: In pregnant women who were carriers of hemophilia A, 32 of the fetuses were male, and DNA examinations were performed in 22 cases. In 16 cases abortions were induced (in 10 cases hemophilia was confirmed by DNA examination), and in 4 of 16 deliveries affected males were born (the disease was confirmed by DNA examination during pregnancy). Of 3 confirmed male fetuses of heterozygous women with hemophilia B, 1 healthy male was born. In 2 cases abortions were induced (in 1 case on the basis of DNA diagnosis). CONCLUSIONS: In cases of heterozygous mothers (hemophilia A and B together) the rate of spontaneous abortions was 13.1%. The rates of premature deliveries (8.2%) and cesarean sections (8.2%) were no higher than national average. The rate of bleeding complications during pregnancy was 18.7%, in 2.7% of cases transfusions were necessary. In case of hemophilic fathers (in heterozygous female fetuses the hemostasis may change from the fetal side) the rate of bleeding complications during pregnancy was 18.2%. In terms of deliveries, obstetrical bleeding complications occurred in 12.2%, atonia in 2%, curettage after delivery in 4.1%, and transfusion in 10.2% of the heterozygous mothers with hemophilia A and B combined. Neonatal complications were cerebral hemorrhage in 1 case and bleeding from the umbilical stump in another case (both newborns were hemophilic males). In connection with delivery, there was no sign of hematoma development on the skull of the newborns, nor were transfusions necessary. In cases of paternal disease the rate of curettage was 6.7% and there were no neonatal or other obstetrical complications. PMID- 12566771 TI - Prenatal evaluation of bidirectional epignathus: comparison of ultrasonography and magnetic resonance imaging. AB - Epignathus is an unusual, benign, congenital teratoma of the hard palate. Most of these teratomas are unidirectional and protrude through the mouth. Hence, the prognosis depends on the size of the tumor and degree of face distortion and airway obstruction. Occasionally, intracranial extension of the tumor is present, involving and destroying the brain tissue, resulting in a poor prognosis. The authors describe 2 cases of bidirectional epignathus, showing the different findings on ultrasonography and magnetic resonance imaging. PMID- 12566772 TI - Prenatal diagnosis of double autosomal mosaicism (47,XX,+8/47,XX,+14): phenotype and molecular cytogenetic analysis on different tissues. AB - A female fetus with multiple congenital anomalies was found to have double autosomal mosaicism, 47,XX,+8/ 47,XX,+14 on chromosome analysis via amniocentesis. At delivery, the proband displayed dysmorphic features of hypertelorism, micrognathia, low set ears, cleft palate, clubfeet, omphalocele, absent gallbladder and congenital heart defects. Fluorescence in situ hybridization demonstrated a marked discrepancy in cell line populations in the tissues examined. PMID- 12566773 TI - Congenital diaphragmatic hernia. Two cases with early prenatal diagnosis and increased nuchal translucency. AB - The early ultrasound prenatal diagnosis of congenital diaphragmatic hernia is uncommon and suggests a poor outcome. We report 2 cases diagnosed at 10 and 12 weeks' gestation, with increased fetal nuchal translucency thickness (4 and 11 mm) and associated abnormalities (complex heart defect in one and many malformations in the other, including duodenal atresia and asplenia). In 1 case, the baby was delivered vaginally at 36 weeks, but neonatal death occurred; the pregnancy was terminated at 15 weeks in the second case. PMID- 12566774 TI - Prenatal diagnosis of a vein of galen aneurysm. AB - We present the prenatal diagnosis of a vein of Galen aneurysm in a 34-week fetus. The aneurysm was 8 x 3.5 x 3 cm in dimension, and located posterior to the third ventricle. Color flow examination revealed a turbulent flow in the lesion. There was significant cardiomegaly. Prenatal ultra-fast magnetic resonance imaging was used to view the malformation in three dimensions. Although the pregnancy continued uneventfully, the fetus died of congestive heart failure on the 3rd day postpartum. The value of antenatal sonography and magnetic resonance imaging in the prediction of prognosis is discussed. PMID- 12566775 TI - Impaired cerebral glucose metabolism in eclampsia: a new finding in two cases. AB - The pathophysiology of the reversible neurological manifestation in eclamptic women remains unclear. We report on 2 women with eclampsia who were repetitively examined by (1) transcranial Doppler (TCD), (2) magnetic resonance imaging (MRI) including T1- and T2-weighted images, fluid attenuated inversion recovery sequence, dynamic susceptibility-weighted perfusion imaging and magnetic resonance angiography (MRA), and (3) (18)fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET). In both cases repetitive TCD revealed no signs of vasospasm; the same was true for MRA. MRI perfusion imaging showed completely homogenous cerebral blood flow in both cases. In the initial phase T2-weighted images revealed hyperintensities in both patients (predominantly bilateral frontal and parietal in 1 and in the temporo-occipital subcortex and the basal ganglia in the other). FDG-PET showed inhomogeneous glucose metabolism (GM) in both patients. Primary increased glucose utilization in the hyperintense T2 weighted areas as well as an attenuated GM parieto-occipital were observed in the 1st case; a high GM was found bilaterally in the basal ganglia and an attenuated one in the occipital cortex in the 2nd. In both cases MRI, and FDG-PET normalized within 3 weeks. These case reports document an altered cerebral GM in the presence of homogenous perfusion in eclamptic women. The high GM may be explained by a decoupling of cerebral perfusion and GM, possibly indicating an increased neuronal activity. The attenuation of the GM is most probably due to a deafferentation of cortical neurons. PMID- 12566776 TI - Sonographic measurement of the fetal iliac angle in trisomy 21, 18 and 13. AB - OBJECTIVE: To determine whether iliac wing angle measurement in second trimester fetuses is a useful sonographic marker for the detection of trisomy 21, 18 and 13. METHODS: During the period between September 1998 and September 2001, 406 fetal iliac angle measurements were performed in women in the second trimester of their pregnancies. The iliac angle measurements in fetuses with trisomy 21 (n = 25), trisomy 18 (n = 10) and trisomy 13 (n = 5) were compared with iliac angle measurement in fetuses with normal karyotypes (n = 333). RESULTS: The mean iliac wing angle in the fetuses with trisomy 21 was 92.67 and 79.35 degrees and 74 degrees in fetuses with trisomy 18 and 13 (the mean iliac wing angle in the healthy fetuses was 70.09 degrees ). CONCLUSION: The proven larger iliac wing angle in neonates with Down's syndrome can be demonstrated sonographically during the pregnancy, especially during the second trimester, and may be useful in prenatal screening of trisomy 21. The sonographic measurement of the fetal iliac angle cannot be used as a marker for trisomy 18 and 13. We have shown that fetuses with trisomy 18 and 13, on average, have iliac angles only a few degrees larger than healthy fetuses. PMID- 12566777 TI - Mathematical models to predict long bone lengths by transvaginal scan at 11-16 weeks' gestation. AB - OBJECTIVE: To propose a new mathematical model to estimate the length of fetal long bones in early pregnancy that can be used in the routine clinical setting. METHODS: In a group of 400 singleton normal fetuses, referred for transvaginal ultrasound examination between 11 and 16 weeks' gestation prior to genetic amniocentesis, a regression analysis was performed to evaluate the relationship between femur length (FLl) (mm) and biparietal diameter (BPD) (mm) and gestational age (GA) (days), as well as between humerus length (HL) (mm) and BPD (mm) and GA (days). The confidence intervals (CIs) of the predicted values for different values of BPD and for different gestational periods and CIs for the regression coefficients are stated as the mean +/- SD of standardized residuals. The accuracy of our best models obtained were evaluated at each gestational week between 11 and 16 with a 10% error cut-off limit. RESULTS: The best relationships between FL and HL versus BPD and GA are: expected FL = -16.92108 + 0.4569402 BPD +0.171617 GA (R(2) = 0.86) and expected HL = -16.28531 + 0.4283019 BPD + 0.1696017 GA (R(2) = 0.88), respectively. When a cut-off limit of 10% in estimating fetal long bones was utilized, the mathematical models revealed a good accuracy particularly at 13-14 weeks' gestation, a period when transvaginal biometric and morphologic examination is advisable and the highest percentage of scans are performed. DISCUSSION: Our proposed two linear models had a very good ability to estimate FL and HL and, due to the simplicity of the calculations, would be particularly useful in the routine clinical setting. PMID- 12566778 TI - Cervical ripening with prostaglandin E1: how an ambulatory method decreases the hospital stay in abortus with intrauterine fetal demise. AB - OBJECTIVE: This study was designed to determine whether use of prostaglandin E(1 )(PGE(1)) is justified to improve the known clinical outcome of prostaglandin E(2) (PGE(2)) gel, because PGE(2) gel preparations are more costly than PGE(1) tablets in most countries, and data to support the use of the gel in clinical practice is not conclusive. The aim was to compare the safety and efficacy of PGE(1) gel when applied in both an in-hospital or ambulatory setting to oxytocin infusion in those women with unfavorable cervical conditions prior to surgical abortion for either medical or obstetrical indications with intrauterine fetal demise. Surgical dilatation of the unripe cervix may result in cervical injury of uterine perforation which could prolong the hospital stay. METHODS: We used PGE(1) gel prepared from tablets and administered in the ambulatory form (group 1), the same PGE(1) gel administered in the labor room (group 2) and intravenously administered oxytocin in the labor room (group 3) for the induction of abortus in women complicated with intrauterine fetus death and missed abortion. Patients requesting abortion were eligible for inclusion, with >8 and <13 weeks of gestation. Eighty-nine women with unfavorable cervices (Bishop score 25% of the maximum capacity was present in 6 patients who performed clear intermittent autocatheterization postoperatively (2 patients preoperatively). In all patients there was a significant improvement in the irritation micturition symptomatology (pollakisuria, nycturia, urgency and urge incontinence) and the quality of life score improved on average from 5 to 0.7. In the case of the patient with renal insufficiency, the creatinine level decreased from 286 to 150 micromol/l; in the other patients renal function remained normal. CONCLUSIONS: Augmentation cystoplasty is a safe and effective method for indicated patients, which significantly enhances their quality of life. PMID- 12566812 TI - Can conventional examinations contribute to the diagnostic power of unenhanced helical computed tomography in urolithiasis? AB - INTRODUCTION: To investigate whether ordinary tests used in urolithiasis can increase the diagnostic value of unenhanced helical computed tomography (HCT). MATERIALS AND METHODS: Of all patients admitted with acute flank pain, 137 without radiocontrast allergy, pregnancy and renal insufficiency were included in the study. After urinalysis (Ur) and ultrasonography (USG), HCT without a contrast agent was taken by 5-mm collimation, thereafter excretory urography (EU) was obtained. Confirmation of a stone was done by various tests and/or treatment methods. Statistically, the predictive values of HCT, USG, EU and Ur and their combinations were evaluated using the area under the receiver operating characteristic curve (AROC). RESULTS: The patients' mean age was 38 (range 4-72) years. Sensitivity values of HCT, USG, EU and Ur were 97, 69, 71 and 81%, respectively. Their specificity values were 96, 87, 91 and 65%, respectively. The widest AROC was found for HCT (97%). The best test combination was [HCT or USG] according to the AROC value (93%), but none of the AROC values of the test combinations was higher than that of HCT. However, 95% confidence intervals of the AROC values of these tests and their combinations overlapped. CONCLUSIONS: Because the HCT combinations with other tests improved the diagnostic value of HCT, they should be preferred in some individuals suspected of having urolithiasis. However, the most feasible single test meeting the all usage purposes was HCT in terms of the AROC value, and the combination was [HCT or USG] in which either HCT or USG was positive. PMID- 12566813 TI - Use of percent free prostate-specific antigen density to improve the specificity for detecting prostate cancer in patients with normal rectal examinations and intermediate prostate-specific antigen levels. AB - INTRODUCTION: As prostatic volume influences percent free prostate-specific antigen (PSA) in patients with prostate cancer, we evaluated whether percent free PSA density, which relates the serum percent free PSA to the volume of the prostate determined by transrectal ultrasound, could increase the specificity of cancer detection in men with normal digital rectal examinations and intermediate PSA levels. MATERIALS AND METHODS: We prospectively evaluated 105 consecutive men with normal digital rectal examinations and serum PSA levels of 4.1-10.0 ng/ml. All patients underwent at least sextant prostate biopsies to establish the diagnosis. For detection of total and free PSA we used an Immulite free and total PSA assay. We compared PSA density, percent free PSA and percent free PSA density for their utility in cancer detection. RESULTS: Overall, 20 of 105 patients (19%) had prostate cancer. The area under the curve (AUC) for percent free PSA density was 0.771, not significantly higher than the AUC of 0.75 for PSA density (p = 0.778), but significantly higher than the AUC of 0.604 for percent free PSA (p = 0.021). Of these three parameters, percent free PSA density yielded the highest specificity percentage (54.1%) at 95% sensitivity. CONCLUSIONS: Percent free PSA density is more specific than percent free PSA in distinguishing benign from malignant disease in men with a normal digital rectal examination and an intermediate PSA level. Further study is necessary to discover whether percent free PSA density is superior to percent free PSA or PSA density. PMID- 12566814 TI - Prognostic significance of p53 nuclear overexpression in patients of muscle invasive urinary bladder carcinoma treated with cystectomy. AB - INTRODUCTION: The objective of the study was to determine the pattern of p53 expression in patients with muscle invasive bladder cancer treated with cystectomy and to assess the prognostic value of p53 expression in this group of patients. MATERIALS AND METHODS: We retrospectively analyzed data of 100 patients who underwent cystectomy for invasive transitional cell carcinoma of the urinary bladder. These patients were seen at 3-monthly intervals during the first 2 years, 6-monthly in the third year, and yearly thereafter. Immunohistochemical analysis for p53 was done on paraffin-fixed tissues with DO-7 antibodies. The tumours with >30% cells staining for p53 proteins were considered p53 positive. RESULTS: There were 95 males and 5 females with a mean age of 54 years. Two patients died in the perioperative period and were excluded from analysis. Over a median follow-up period of 2 years (range 3 months to 10 years) 62 patients are alive without disease while 36 had relapse and of these, 31 have died. The p53 expression was significantly related to stage and grade of the tumour. p53 negative tumours were more likely to be organ confined and of lower grade as compared to p53-positive tumours. The stage of tumour was significantly related to outcome but grade and p53 immunohistochemistry did not provide any prognostic information. CONCLUSIONS: p53 immunostaining was positive in 57% of muscle invasive urinary bladder tumours. The p53 positivity correlated well with the stage and grade of the disease, whereas it has no additional prognostic value. PMID- 12566815 TI - Stage pT1 renal cell carcinoma: review of the prognostic significance of size. AB - INTRODUCTION: The last TNM classification (before 1997) defined T1 kidney tumour as a tumour <2.5 cm, limited to the kidney: this cut-off point was changed in 1997 and T1 neoplasm was defined as <7 cm. This new cut-off now includes past T1 and T2 diseases, without any differences in terms of prognoses. We performed a retrospective analysis of our cases and specifically investigated if there were any differences in terms of prognosis in a group of patients, currently identifiable as pT1, if they were divided into two subgroups based on tumour pathological sizes. MATERIALS AND METHODS: Our analysis involved 128 patients (mean age 57.6 years) who underwent radical nephrectomy or nephron-sparing surgery in the period between 1990 and 2000. All these patients were pT1 according to the new TNM classification and were divided into two groups according to different cut-off point sizes (from 2.5 to 5 cm). We analysed the surgical approach, overall survival and cancer-specific mortality in the two subgroups, renamed as pT1a and pT1b, and performed a statistical analysis of the results using the Kaplan-Meier method to prove if this substaging identified changes in survival outcome. RESULTS: We obtained more interesting results for a 5-cm cut-off: the two groups showed a similar follow-up and overall survival rate but different cancer-specific mortality rate (6 vs. 12.1%). The statistical analysis showed that the two survival curves (pT1a vs. pT1b disease) had a similar trend up to about 60 months; after this period the two curves diversify with a drop in survival rate among patients with larger tumours (pT1b patients). CONCLUSIONS: It would seem reasonable to reassess the TNM classification of stage pT1 in order to better define prognosis in this group of patients. PMID- 12566816 TI - Endovascular repair of abdominal aortic aneurysm in renal transplantation. AB - Successful endovascular correction of a 12-cm abdominal aortic aneurysm (AAA) is described in a 76-year-old man with a functional pelvic renal transplant and 18 month follow-up. Endovascular treatment is a safe alternative to surgery for AAA correction in the elderly post-transplantation patient since it does not require flow interruption during the procedure. PMID- 12566817 TI - Pyruvate kinase type tumor M2 in urological malignancies. AB - INTRODUCTION: The dimeric form of pyruvate kinase type M2 is overexpressed in tumor cells (TuM2-PK). The aim of the present study was to evaluate the clinical value of TuM2-PK as a tumor marker for renal cell carcinoma (RCC), transitional cell carcinoma of the bladder (TCC) and prostate cancer (PCA) by using a commercially available enzyme-linked immunosorbent assay for detection of TuM2-PK in plasma. MATERIAL AND METHODS: The TuM2-PK concentration in EDTA plasma was determined quantitatively and immunologically using an ELISA (ScheBoTech, Germany). We measured the TuM2-PK plasma levels of 83 patients with RCC, 30 patients with TCC and 30 patients with PCA before any therapy. 100 patients with various non-malignant urological disorders were recruited as the control group. RESULTS: Only patients with RCC showed significantly elevated plasma levels of TuM2-PK compared to the control group (p < 0.01). We found a sensitivity of 42.6% and a specificity of 80.4% using a cut-off value of 15 U/ml (manufacturer's recommendation). During follow-up, only 50% showed increasing plasma levels of TuM2-PK in case of metastases. Significant differences could not be detected in either TCC or PCA. CONCLUSIONS: Our data suggest that TuM2-PK is not a useful marker for TCC and PCA. Due to low sensitivity and specificity, TuM2-PK is not suitable for the diagnosis of RCC. Whether TuM2-PK may be useful in advanced RCC to control success of palliative treatment regimens is still unclear. PMID- 12566818 TI - Analysis of premature ejaculation in hemodialysis patients using the International Index of Erectile Function. AB - OBJECTIVE: We have conducted the following study to determine the prevalence of premature ejaculation in male hemodialysis patients and its impact on sexual quality of life. METHODS: This study was performed in hemodialysis facilities. Patients were asked to complete the International Index of Erectile Function (IIEF) questionnaire and also to report their ejaculation status. 98 male hemodialysis patients with normal potency were included in the study. The IIEF domain scores were determined, and the overall and relationship satisfaction rates were calculated for the patients with and without premature ejaculation separately. RESULTS: The mean age was 45.9 (range 30-69) years and the duration of hemodialysis was 37.8 (range 7-84) months. Premature ejaculation was determined in 31 (31.6%) patients. Patients with premature ejaculation experienced significantly lower overall satisfaction scores. Items of satisfaction with overall sexual life and sexual relationship with partner were responded to with a score of 'moderately satisfied' or 'very satisfied' in 47 and 53% of patients with premature ejaculation and 84 and 86% of patients without premature ejaculation, respectively. CONCLUSIONS: Premature ejaculation is a prevalent disorder in hemodialysis patients and seems to affect the satisfaction rate and sexual quality of life of these patients. PMID- 12566819 TI - Effects of castration on nitric oxide-mediated relaxations in male rat corpus cavernosum smooth muscle. AB - BACKGROUND: The effects of castration on nitric oxide- mediated relaxations and nitric oxide synthase activity in male rat corpus cavernosum smooth muscles. METHODS: Eight-week-old male rats were assigned to two groups: control (sham operated) and castrated animals. After 8 weeks, corpus cavernosum smooth muscle strips were mounted in an organ bath for isometric tension recordings. Electrical field stimulation (EFS) was applied to the strips precontracted with 30 microM phenylephrine. The microdialysis probe was inserted into the strip, and Krebs Henseleit solution was perfused into the probe. The dialysate during EFS and cholinergic stimulation was collected, and the amount of NO(-)(2)/NO(-)(3) (NOx) released in the dialysate was measured by the Greiss method. Sodium nitroprusside and carbachol were cumulatively added to the strips precontracted with 30 microM phenylephrine. RESULTS: EFS caused frequency-dependent relaxations and NOx releases in the strips. Pretreatment with N(omega)-nitro-L-arginine (100 microM) and tetrodotoxin (1 microM) completely inhibited relaxations and NOx releases. The maximum relaxation in the castration group was significantly greater than that in the control group. The release of NOx was significantly greater in the castration group than in the control group. Sodium nitroprusside relaxed the tissues in both groups similarly. Carbachol failed either to relax the tissue or to increase the amount of NOx production in the tissue. CONCLUSION: The present data suggest that castration enhances nitric oxide synthase activity and nitric oxide-mediated relaxations in the male rat corpus cavernosum. PMID- 12566820 TI - Infestation of the human kidney with Dioctophyma renale. AB - Human infestation with Dioctophyma renale is presented. Clinical signs and diagnostic findings are unspecific. They are discussed and a conservative therapeutic approach is suggested. PMID- 12566821 TI - Distal ureteral stenosis after early adjuvant intravesical mitomycin C application for superficial bladder cancer. AB - We report a case of distal ureteral stenosis after transurethral resection of a small bladder tumor near the left ureteral orifice and early postoperative mitomycin C instillation for prevention of recurrence. The patient developed late recurrent stenosis of the ureteral orifice with histologic evidence of localized, severe benign inflammatory reaction. The recurrent stenosis was successfully managed by transurethral resection of the scar tissue and ureteric stenting. Although ureteral stenosis does occur after transurethral resection, the severity and time course of the stenosis in this case suggest an influence of the intravesical chemoprophylaxis used. PMID- 12566822 TI - Spontaneous rupture of an aneurysmal intrarenal arteriovenous fistula. AB - We present a 65-year-old woman who was found to have a spontaneous rupture of an aneurysmal intrarenal arteriovenous fistula. To our knowledge, the present case is only the 3rd reported in the literature. The preferred method of treatment of ruptures of arteriovenous fistulae is embolization. However, the process of choosing among embolization, surgery, or a combination of both procedures must be individualized for each patient based on vital signs and symptoms. PMID- 12566823 TI - Efficacy measurement of topical antihistamines: a review. AB - More and more antihistamines are used in gels or ointments for local antipruritic therapy. Among other factors, the efficacy is dependent on the penetration properties of the respective agents and the optimal choice of vehicle substances. To avoid expensive treatment with unsatisfying success, a reliable efficacy measurement would be desirable prior to the admission of new topical antihistamine preparations. Therefore we reviewed the literature for common methods to assess the efficacy of local antihistamines in healthy volunteers. The principle is to apply the test substance to marked test areas and to challenge the skin after a certain time with a standardised amount of histamine, allergens or mast-cell-degranulating substances. For the test evaluation, the areas of wheal, flare and itch are measured and compared between antihistamine-treated and control fields. Challenge models and most of the described evaluation methods are suited for the preliminary efficacy measurement of antihistamines. However, to be able to compare the results, a standardised procedure used by all investigators would be desirable. PMID- 12566825 TI - In vivo tolerance assessment of skin after insertion of subcutaneous and cutaneous microdialysis probes in the rat. AB - The purpose of the study was to evaluate the trauma induced by insertion of the linear microdialysis probe in the subcutaneous and dermal tissue in the rat and to check if the microdialysis probe insertion affects transdermal drug delivery. Non-invasive bioengineering methods (TEWL, Laser Doppler Velocimeter, Chromameter) as well as histology were combined to characterize these effects. The results showed that the dermal and subcutaneous insertion of microdialysis probes did not change skin permeability, blood flow and color, confirming the safety of this technique. The probe depth did not influence the trauma. No significant physical damage after probe insertion was noticed. Thus, the present work validates the use of microdialysis in dermatopharmacokinetics studies after topical or systemic drug delivery. PMID- 12566824 TI - The effects of topical corticosteroids and a coal tar preparation on dithranol induced irritation in patients with psoriasis. AB - Dithranol has been a mainstay in the treatment of psoriasis for more than 80 years. Although a safe approach, the irritation of the clinically uninvolved perilesional skin remains a major limitation of this treatment. Corticosteroids and coal tar solution have an anti-inflammatory potential. The aim of the present study was to investigate the clinical and cell-biological effects of two topical corticosteroids and a coal tar preparation on dithranol-irritated skin. During 4 consecutive days, 2% dithranol cream was applied to six uninvolved skin sites (3 cm in diameter) on the lower back of 9 patients with psoriasis. Dithranol was left on the skin for 1 h, subsequently removed with water and soap and the skin was dried with a towel. Subsequently, SITE 1 was treated with 0.05% clobetasol-17 propionate ointment (CP), SITE 2 with unguentum cetomacrogolis (vehicle 1), SITE 3 with 0.005% fluticasonpropionate ointment (FP), SITE 4 with 10% coal tar solution in lanettewax cream (CTS), SITE 5 was left untreated (control) and SITE 6 was treated with lanettewax cream (vehicle 2). Erythema, oedema and vesicle formation was scored every day. On day 5, punch biopsies were taken from the six sites. The expression of epidermal proliferation, differentiation and inflammation markers and the clinical irritation scores indicate that the application of a high potency corticosteroid (CP) is the best approach to minimise dithranol irritation, whereas CTS had virtually no effect on dithranol irritation during this 4-day experimental model. PMID- 12566826 TI - Skin penetration and sun protection factor of five UV filters: effect of the vehicle. AB - To gain information about efficacy and safety of sunscreens, we compared the skin penetration of ultraviolet (UV) filters from two vehicles, i.e. an oil-in-water (O/W) emulsion gel and petrolatum jelly both in vitro and in vivo, as well as the corresponding pharmacological effect, i.e. the sun protection factor (SPF) in vivo. The UV filters studied were benzophenone-3 (BPH), ethylhexyl methoxycinnamate (EHM), butyl methoxydibenzoyl methane, ethylhexyl salicylate and homosalate. The human skin penetration of these five chemicals from the two vehicles was determined both in vitro using Franz cells and in vivo using a standardized tape-stripping method. The SPF of the two sunscreens was determined in vivo following the COLIPA guidelines. In vitro none of the filters permeated through the skin after 6 h of product application and very little could be found in the skin. BPH and EHM were the only UV filters found in the dermis (both after 30 min and 6 h). An effect of the vehicle could be noticed only for BPH after 30 min in the dermis and 6 h in both dermis and epidermis. In vivo, no differences in the amount of individual UV filters (in % of the applied dose) in the 15 first strips of the stratum corneum (SC) were found following 30 min of application of the formulations; however, the amount of UV filters that were retained in the SC was significantly higher (around 3 times) with the O/W emulsion gel than with the petrolatum jelly. This difference between the two vehicles was also of consequence for the SPF in vivo measured 30 min after application of the products (SPF congruent with 18 with the O/W emulsion gel compared to SPF congruent with 10 with the petrolatum jelly). By choosing the right vehicle or optimizing it, not only sunscreen products can be significantly improved in terms of pharmacological efficacy but the potential toxicological risk associated with the skin penetration of UV filters may be significantly reduced. PMID- 12566827 TI - Beneficial effects of intraperitoneally administered alpha-tocopheryl acetate on the levels of lipid peroxide and activity of glutathione peroxidase and superoxide dismutase in skin, blood and liver of thermally injured guinea pigs. AB - The aim of this study was to investigate the effects of intraperitoneal alpha tocopheryl acetate administration on concentration of lipid peroxide (as malonyldialdehyde, MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in plasma, red blood cells (RBC), skin and liver of thermally injured guinea pigs. MDA levels in the plasma, skin and liver were increased at the 15th min after the thermal injury compared to the control group. However, they were significantly lower in the alpha-tocopheryl-acetate treated groups than both control and thermally injured groups from the 15th min to the 6th day. SOD activity of the skin and RBC and vitamin E levels in the plasma were significantly decreased between the 15th min and 6th day and then both parameters, i.e. SOD and vitamin E, were significantly increased by alpha tocopheryl acetate administration. GSH-Px activity in the skin was also significantly decreased in groups from the 15th min to the 6th day, whereas it remained unchanged by the alpha-tocopheryl acetate administration. On the other hand, GSH-Px activity and GSH levels in the RBC remained the same during both thermal injury and alpha-tocopheryl acetate administration. In conclusion, we observed that the intraperitoneal administration of alpha-tocopheryl acetate protects the skin against thermal injury, which suggests that it may be due to the upregulation of enzymatic antioxidants. PMID- 12566828 TI - Peripheral sensory neuropathy associated with short-term oral acitretin therapy. AB - A 57-year-old female patient with widespread chronic plaque psoriasis and a 32 year-old male patient with severe oral lichen planus are reported, who developed sensory symptoms in the extremities 3 and 4 months after the onset of oral acitretin therapy, respectively. Both patients showed clinical and electrophysiological evidence of a sensory peripheral neuropathy, which completely resolved 2 and 2.5 years after discontinuation of oral acitretin administration, respectively. PMID- 12566829 TI - Alteration of the expression of Bcl-2, Bcl-x, Bax, Fas, and Fas ligand in the involved skin of psoriasis vulgaris following topical anthralin therapy. AB - Anthralin (dithranol) is frequently used for the treatment of psoriasis. However, the mode of action of anthralin has not been completely elucidated as yet. Recent findings suggest that psoriatic keratinocytes are resistant to the apoptotic process. In this study, we examined the immunohistochemical expression of apoptosis-regulated protein in the involved psoriatic skin following topical anthralin therapy. Biopsy specimens were obtained from back skins treated with topical anthralin or white petrolatum (control) in 4 patients with psoriasis vulgaris. Immunohistochemical examination revealed that psoriatic keratinocytes expressed high levels of Bcl-x, which was significantly reduced after anthralin treatment. Bax was not detected in the epidermal keratinocytes in the petrolatum treated skin, while it was present in the upper keratinocytes after anthralin therapy. Bcl-2 was detected only in basal layers of psoriatic epidermis following both petrolatum and anthralin application. Psoriatic keratinocytes expressed higher levels of Fas in the lower epidermis, while only weak expression was detected in anthralin-treated plaques. On the other hand, hyperproliferative keratinocytes strongly expressed Fas ligand (FasL) on their plasma membranes as well as infiltrating lymphocytes in the upper dermis. Furthermore, anthralin treated psoriatic epidermis did not express FasL. In normal skin, keratinocytes expressed low to absent levels of Bcl-x and Bax, while Bcl-2 was detected only in melanocytes in basal layers. Neither Fas nor FasL were detected in the epidermis of normal skin. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) staining revealed positive labeling on the majority of psoriatic keratinocytes through the epidermis in petrolatum-treated skin, whereas anthralin treatment markedly reduced TUNEL-positive keratinocytes. These in vivo results may reflect improvement of the psoriatic skin following effective anthralin therapy. PMID- 12566830 TI - Reactivity of contact allergenic haptens to amino acid residues in a model carrier peptide, and characterization of formed peptide-hapten adducts. AB - The type of chemical reaction between hapten and carrier protein in the formation of a complete antigen in vivo giving rise to an allergic contact dermatitis (ACD, type IV allergy) is essentially unknown. About 4000 low-molecular organic compounds are known to have allergenic properties. alpha,beta-Unsaturated carbonyl structures are frequently present among these compounds. Haptens giving rise to antibody formation and type I allergy have been shown to add predominantly to lysine in the carrier protein. In this paper, the reactivity of activated type IV haptens to a model peptide is reported. Essentially all amino acids with nucleophilic properties were present in the model peptide. Investigation of the relative reactivities of the amino acid residues to activated haptens under biomimetic conditions is performed in order to determine the proportions between the adducts of the different amino acid moieties. In all cases, the electrophilic alpha,beta-unsaturated haptens were found to be added to the cysteine residue and no lysine adduct was recorded. Nuclear magnetic resonance (NMR) spectroscopy was used to exclude steric hindrance of any amino acid residue in the addition reaction. The hapten-modified peptides were isolated and characterized by NMR and mass spectrometry. PMID- 12566831 TI - MRI-guided frameless stereotactic percutaneous cordotomy. AB - BACKGROUND: Use of intraoperative myelography as a radiologic guidance for percutaneous cervical cordotomy (PCC) has been superseded by more modern imaging. The only significant advancement in cordotomy techniques over the last 30 years has been CT-guided PCC. The goal of this study was to demonstrate the feasibility of an MRI-guided frameless technique in high cervical cordotomy. METHODS: We describe 6 patients with intractable pain treated using a frameless, MRI-guided, stereotactic, PCC technique in combination with standard physiological localization procedures. Results were compared with those from 32 patients who underwent PCC in the last 5 years using physiological localizing techniques only. RESULTS: Six patients (100%) who underwent the frameless technique had excellent pain relief postoperatively. Patients in the non-stereotactic group, on average, required a higher number of lesions (2.5 vs. 1.2, p < 0.005), and 7 (22%) of these patients had unsatisfactory pain relief following PCC (p = 0.21). Five patients in the non-stereotactic group had weakness postoperatively and 1 had changes in bladder function. Postoperative weakness occurred in 1 patient undergoing the frameless technique. At an average of 6 months of follow-up (range 5-11), excellent pain relief was achieved in 83% (5/6) of MRI frameless PCC patients and 55% (16/29) of standard PCC patients (p = 0.20). CONCLUSIONS: Intraoperative frameless stereotaxy provides surgeons with accurate information that helps to guide the operative approach and precisely tailor the trajectory and depth of the electrode, potentially increasing the safety and efficacy of the operation. PMID- 12566832 TI - Impact of posterior GPI pallidotomy on leg tremor in Parkinson's disease. AB - Although stereotactic thalamotomy is the mainstay in the surgical treatment of tremor in patients with Parkinson's disease (PD), this surgery is not favored and is even a matter of potential concern in the treatment of leg tremor since it carries a significant risk of injury to the internal capsule. In this study we have carried out a quantitative assessment of leg tremor alleviation in 12 patients with PD after MRI-/microelectrode-guided stereotactic ablation of the posterior part of the globus pallidus internus (GPi). The results showed that posterior GPi pallidotomy combined with drug therapy is a satisfactorily effective therapeutic strategy to treat parkinsonian leg tremor. PMID- 12566833 TI - Brain metastasis: experience of the Xi-Jing hospital. AB - BACKGROUND: The management of metastatic brain tumors is an important issue in patients with malignant tumors or cancer. The authors summarize the results of patients with brain metastases treated at the Xi-Jing Hospital during a 10-year period, in order to assess the best modality of treatment for patients with brain metastases. METHODS: Between 1990 and 2000, 463 patients with brain-metastatic tumors were treated at the Xi-Jing Hospital of the Fourth Military Medical University, Xian, China. In most patients, the pathologic diagnosis of primary cancer was obtained before they were referred for their brain metastasis. There were 34 (8.42%) cases with an unknown primary cancer site at the time of initial presentation. Patients were grouped according to treatment methods, which included neurosurgical craniotomy (NS; 130 patients), whole-brain radiotherapy (WBRT; 120 patients). Linac XKnife radiosurgery (RS; 130 patients) and Linac XKnife radiosurgery plus WBRT (RT; 83 cases). Survival was measured from the time of treatment and was analyzed by the Kaplan-Meier product-limit method and then plotted. Differences between curves were evaluated using the log-rank test. Multivariate factors associated with survival were analyzed using the Cox proportional hazards model. RESULTS: The survival time was 68.4 +/- 7.20 weeks after NS, 51.3 +/- 5.04 weeks after WBRT, 67.9 +/- 3.68 weeks after RS and 89.7 +/- 4.50 weeks after RT. The presence of active systemic cancer in a larger number of metastatic tumors was associated with a poor survival (p = 0.0003 and 0.0000). The female patients showed better survival rates over the male ones (p = 0.0000). Patients treated with RT had a better survival than those treated with NS, WBRT and RS (p = 0.0048, 0.0000 and 0.1222, respectively), although the latter did not show statistical significance. CONCLUSIONS: RS was an effective modality for patients with brain metastases, and if combined with WBRT, survival was better. Progression of systemic cancer and the number of metastatic tumors were the most significant factors for a poor survival after treatment of the brain metastases. PMID- 12566834 TI - Preliminary exploration of the clinical effect of bleomycin on craniopharyngiomas. AB - OBJECTIVE: To investigate the antitumor effect of bleomycin on craniopharyngiomas. METHODS: A series of cystic craniopharyngiomas were randomly divided into three groups: (A) intracystic chemotherapy with bleomycin; (B) intracystic chemo-radiotherapy with bleomycin and (32)P; (C) intracystic radiotherapy with (32)P and 0.9% saline. The agents were injected into the cysts through stereotactically inserted silicone tubes. Follow-up was done for a minimum of 6 months. Outcome was based on a comparison of the volume of cysts before treatment and at follow-up. The index and lactate dehydrogenase (LD) of the cystic fluids, blood and cerebrospinal fluids and the endocrine function of these patients were determined before and after therapy. RESULTS: 19 patients finished the whole therapeutic course: 5 from group A, 9 from group B and 5 from group C. Four tumors in group A were polycystic, and the drug was selectively injected into the largest cyst. At follow-up, the volumes of the cysts in groups A and B regressed from 92 to 0%, while the drug-free cysts enlarged. In group B, 6 cysts almost disappeared and another 3 regressed from 78 to 57%. In group C, one cyst progressed and the others shrank by different degrees, but none disappeared completely or nearly. All patients in groups A and B had fever of different degrees, which resolved spontaneously in 8-24 h. The complications in group B included hyponatremia in 1 patient, and both adephagia obesity and cerebral infarction in 2 patients (1 of whom died after 6 months). Apart from the oculomotor paralysis occurring in 1 patient, the remainder of group C had no other severe complications. Blood chemistry, liver, kidney, pituitary and endocrinal functions changed little during the course in all these 19 patients. LD and its isoenzymes from the cystic fluids, CSF and serum showed no marked change after bleomycin injection. CONCLUSION: Bleomycin injected into cysts of craniopharyngiomas causes the tumor to shrink. When (32)P is added, the therapeutic effect seems better than treatment with either (32)P or bleomycin alone. Blood chemistry, liver, kidney and endocrine functions change little irrespective of the therapy applied. However, the combination of chemotherapy and radiotherapy may severely disturb both serum electrolytes and endocrine function. LD and its isoenzymes in the cystic fluids, CSF and serum may not change after bleomycin treatment. PMID- 12566835 TI - Irving S. Cooper and his role in intracranial stimulation for movement disorders and epilepsy. AB - Dr. Irving Cooper (1922-1985) was a pioneer in the field of functional neurosurgery. After years of treating patients with tremor by creating deep lesions with either anterior choroidal artery ligation or cryogenic thalamotomy, he began to utilize methods of electrical cerebral stimulation as treatment for a variety of disorders. Chronic cerebellar stimulation was employed for patients with epilepsy, cerebral palsy, and dystonia. While Dr. Cooper believed his results to be significant, there still remain many challenges to his claims. Later in his career, he placed deep brain electrodes in the internal capsule and thalamus for epilepsy and dystonia. His encouraging results from this small series were often overlooked. This paper reviews the science behind Dr. Cooper's work, his case series, and the controversies surrounding his results. PMID- 12566836 TI - A systematic review of brachial plexus surgery for birth-related brachial plexus injury. AB - OBJECTIVE: Brachial plexus injury complicates 0.6-2.6 per 1,000 live births. Surgery is sometimes advocated for patients who fail to improve with conservative management. We reviewed the available literature on birth-related brachial plexus palsy in order to provide recommendations for surgical management, using evidence based criteria. METHODS: Studies were identified by searching Medline (1966-2002) and the Cochrane Library using the terms brachial plexus, neonate or infant, surgery and natural history. The reference lists of relevant articles were also reviewed. The search was restricted to articles published in English. Each article was classified according to its methodology. Management recommendations were suggested based on the results of the studies identified and the degree of certainty of the available literature. RESULTS: Twenty-three papers were selected for detailed analysis. There are no randomized controlled trials that have investigated the role of brachial plexus surgery in the management of birth related brachial plexus palsy. Two prospective studies of relevance were found, one describing the natural history of birth-related brachial plexus injury and one evaluating surgery for these patients. The remainder consisted of retrospective case series. Outcomes from surgical series are generally favorable (level III and V evidence). Direct comparison with the natural history could not be inferred from the series reviewed given the lack of controls. CONCLUSION: There is no conclusive evidence showing a benefit of surgery over conservative management approaches in the treatment of patients with birth-related brachial plexus injuries. Surgery remains a valid practice option given the level III and V evidence suggesting a possible benefit of surgery. PMID- 12566837 TI - Primary excision of pediatric posterior fossa abscesses--towards zero mortality? A series of nine cases and review. AB - Nine children with infratentorial brain abscesses were treated at our institution over a 3-year period. Seven of them were boys and two were girls. In five cases, the abscesses were otogenic, and in two cases, the cause was tubercular (including one case of tubercular otitis). There were three cases of abscess formation in dermoids. All cases were managed with primary excision of the abscess. The average hospital stay was 12 days, and there was no mortality. The most common complication was hydrocephalus, noticed in three patients, of whom one eventually required shunt placement. Our policy of primary excision of these abscesses has yielded gratifying results. This approach also resulted in minimal morbidity and a substantially shortened hospital stay. Moreover, primary excision yields definitive histopathology of the lesion, especially in cases of tubercular and dermoid etiologies. PMID- 12566838 TI - Evaluation of cerebral hemodynamics with perfusion MRI in childhood moyamoya disease. AB - The purpose of our study was to evaluate the role of perfusion MRI and to compare the findings with those of conventional imaging in childhood moyamoya disease (MMD). Perfusion MRI was performed preoperatively on 13 cases of childhood MMD. Based on the perfusion MRI findings, patients were classified into four groups: those with normal regional cerebral blood volume (rCBV) and time to peak (TTP) (group 1, n = 2); those with normal rCBV and delayed TTP (group 2, n = 1); those with increased rCBV and delayed TTP (group 3, n = 5), and those with decreased rCBV and delayed TTP (group 4, n = 5). Lesion volume as determined using the rCBV map was compared with that on MRI and single photon emission computerized tomography (SPECT). The characteristics of groups 1, 2 and 3 were a high frequency of transient ischemic attacks, normal finding on MRI, early angiographic stages, decreased reserve on SPECT and favorable operative outcome. In group 3, angiography revealed abundant collateral channels. In group 4, all cases had infarction on MRI and abnormal perfusion on SPECT. Angiographic stages tended to have progressed, and operative outcome was less favorable. Perfusion MRI documented hemodynamic asymmetries in the five cases with normal MRI. Perfusion MRI is suggested to have a sensitivity intermediate between rest and acetazolamide SPECTs. We conclude that perfusion MRI may be a valuable tool for characterizing and monitoring ischemia in MMD. Perfusion MRI provides additional functional information not available from conventional MRI and has a potential role comparable to SPECT in the evaluation of MMD. PMID- 12566839 TI - Primitive neuroectodermal tumor arising in long-standing cerebellar atrophy. AB - Primitive neuroectodermal tumors (PNETs) account for one fifth of childhood brain tumors. Although little is known of the pathobiology of this tumor type, there are associations with both genetic syndromes and exposures to specific environmental agents. Progressive cerebellar atrophy predating the presentation of a primary brain tumor, in the absence of a genetic syndrome, has not been reported with PNETs. We report a case of a posterior fossa PNET occurring in association with long-standing cerebellar atrophy without evidence of a genetic syndrome. This case may represent an unrecognized paraneoplastic syndrome or a unique subtype of PNET. PMID- 12566840 TI - Tethered cord associated with anorectal malformation. AB - Ten children with a tethered cord and also an anorectal malformation are reported in this document. The anorectal malformations comprised 5 vesicointestinal fissures, 2 cloacal exstrophies, 2 rectovesical fistulas and 1 rectobulbar fistula. All of the patients underwent colostomy in advance of surgery for untethering of the spinal cord. Although their neurologic deficits had previously been considered static, they were subjected to radiographic examination of the caudal spine and found to have a tethered cord. These 10 children were among 55 children with a tethered cord surgically treated at the Division of Neurosurgery of the Osaka Medical Center and Research Institute for Maternal and Child Health during the last 11 years. Data were obtained for these 10 children (6 boys and 4 girls, mean age 1.7 years) who underwent surgical untethering. Several hypotheses are offered to explain this association. Anorectal malformations are related to underlying spinal cord anomalies, which may be amenable to neurosurgical correction. Eight of our patients had no skin stigma of the lumbosacral region, in contrast to an ordinary tethered cord. Spinal cord imaging is necessary to closely scrutinize these children. PMID- 12566841 TI - Intraoperative low-field magnetic resonance imaging in pediatric neurosurgery. AB - BACKGROUND: Since the mid-1990s, the feasibility and indications of intraoperative magnetic resonance (MR) imaging have been investigated by different groups. The majority of examinations were carried out in adults. The aim of this study was to summarize our experience of over 5 years of intraoperative MR imaging in pediatric neurosurgery. METHODS: For scanning, we used a 0.2-Tesla Magnetom Open, which was placed in a radiofrequency-shielded twin operating theater, allowing surgery with standard instruments and additional neuronavigational guidance either in an adjacent operating room or directly in the radiofrequency cabin on the extended MR table, at the 5-G line. RESULTS: In total, 330 patients were investigated, among them 33 children who were younger than 17 years. We found four main indications for intraoperative MR imaging: the evaluation of cyst drainage (n = 9), of the extent of resection in epilepsy surgery (n = 6) and of the removal of pituitary tumors (n = 6) and gliomas and other brain tumors (n = 12). Intraoperative MR imaging allowed us to evaluate the extent of the resection or to monitor catheter placements and consecutive cyst alterations in all cases. In 2 tumor cases and 3 catheter placements, intraoperative imaging resulted in a modification of the surgical strategy. CONCLUSIONS: Intraoperative low-field MR imaging is a safe procedure; we did not encounter an increased morbidity in the children investigated. It serves as intraoperative quality control documenting the effects of surgery, e.g. the extent of a resection, which can then be compared to the treatment plan. Besides its most essential application in brain tumors, it also proved to be particularly helpful in children undergoing complicated catheter placements for cyst drainage, as well as in pituitary and epilepsy surgery. PMID- 12566842 TI - The Seldinger technique for insertion of difficult to place ventricular catheters. AB - BACKGROUND: Multiloculated ventricular hydrocephalus is a difficult pathology for neurosurgeons to treat. Not only are the shunts for this condition prone to malfunction, but they are also difficult to place. METHODS: The Seldinger technique is used in combination with a rigid scope with a working channel. A soft filiform wire or a thick suture is inserted through the channel. The scope is withdrawn and a ventricular catheter is slid down the wire. The remainder of the shunt is inserted in the usual fashion. RESULTS: We have not encountered any neurologic compromise from this technique. No bleeding was encountered with the described technique. CONCLUSION: The Seldinger technique with appropriate types of guide wires and in combination with an endoscope is a useful technique for difficult to place shunts. PMID- 12566843 TI - Long-term follow-up of direct heart shunts in the management of hydrocephalus. AB - The authors performed a retrospective review of clinical data on 60 patients treated with direct heart shunts (DHSs) for hydrocephalus at the Children's National Medical Center from 1976 to 1979. Mean shunt survival was 79 months. Rates of proximal revision and infection were comparable to those in previous shunt series. Distal revision rates were lower than those reported in previous ventriculoatrial shunt series. The DHS is an effective self-lengthening cerebrospinal fluid shunt that may be considered as a salvage technique in the challenging shunt-dependent patient. PMID- 12566844 TI - Extraventricular intracisternal obstructive hydrocephalus--a hypothesis to explain successful 3rd ventriculostomy in communicating hydrocephalus. AB - The basis of successful 3rd ventriculostomy in cases of communicating hydrocephalus is not yet understood. We performed 3rd ventriculostomies in 5 patients with free cerebrospinal fluid (CSF) communication from the ventricles to the cisterna magna. Preoperative magnetic resonance images (MRIs) showed dilated ventricles, a downward bulging floor of the 3rd ventricle (interpreted as a sign of pressure gradient between the ventricles and basal cisterns) and a free communication to an enlarged cisterna magna. The other basal cisterns were of normal or smaller size. All patients recovered from their clinical symptoms and none of them needed a shunt. The hypothesis of an intracisternal CSF pathway obstruction (e.g. between the cisterna magna and the prepontine cistern) could explain the MRI findings, although such an obstruction cannot be directly visualized. It would also explain the successful 3rd ventriculostomies in these cases. PMID- 12566845 TI - Congenital thoracic cord segmental amyelia: a rare manifestation of segmental spinal dysgenesis. AB - Segmental spinal dysgenesis (SSD) is a rare congenital abnormality in which a segment of the spine and spinal cord fails to develop properly. We report a case of a 4-year-old female child who was born prematurely with a permanent neurological deficit in the lower limbs and sphincter incontinence. Radiological investigations revealed segmental agenesis of the thoracic spinal cord (D8-D11 vertebral levels) with an associated vertebral bony anomaly. The pathogenesis of this rare form of SSD syndrome is discussed and the relevant literature is briefly reviewed. PMID- 12566846 TI - The role of surgical placement and pump orientation in intrathecal pump system failure: a technical report. AB - Intrathecal pump catheter complications are the most common cause of failure in drug delivery. A previous report has documented that intra-abdominal positioning of the intrathecal pump may predispose the pump-catheter neck to premature catheter breakdown and leakage. Based on this report, we reviewed over 100 intrathecal pump cases to determine the frequency of malpositioning and its role in the pathogenesis of catheter failure. We found three specific cases where a 'fulcrum effect' occurred due to intra-abdominal positioning of the pump predisposing the catheter to breakdown. This study demonstrates that intra abdominal placement of the pump can predispose the catheter to failure/breakdown and that surgeons should attempt to place the pump catheter neck in a superiomedial position, distant from any bony prominences, to prevent the 'fulcrum effect' on the pump-catheter neck junction and reducing the likelihood of either internal or external compressive forces. PMID- 12566847 TI - Shunt malfunction presenting with cerebral edema. PMID- 12566848 TI - Trypanosoma cruzi: in vitro morphological alterations induced by actinomycin D. AB - Actinomycin (ActD) is an antibiotic that binds DNA, preventing transcription. When a Trypanosoma cruzi infection in mice is treated with this drug, the parasite loses its ability to multiply, enabling protection. In this study, axenic cultured T. cruzi parasites were exposed to different concentrations of ActD (10, 20, and 50 microg/ml), all of them being able to inhibit growth and to alter the mobility. Nevertheless, the parasites remained alive and motile for at least 14 days. Scanning electron microscopy of trypomastigotes treated with 10 microg/ml of ActD for 24 h showed a modification in their morphology which suggests a change in the parasite cytoskeleton. PMID- 12566849 TI - Ethanol reduces cardiac myocyte function through activation of the nitric oxide cyclic GMP pathway. AB - We tested the hypothesis that low-dose ethanol would reduce cardiac myocyte function through increased production in the nitric oxide/cyclic GMP signal transduction pathway, rather than reduced degradation. Ventricular myocytes were isolated from the hearts of 9 rabbits. Myocyte function was studied using a video edge detector and cyclic GMP levels were measured by radioimmunoassay. Cells were administered 5 and 10 mmol/l ethanol alone or after 10(-6) mol/l N(G)-nitro-L arginine methyl ester (L-NAME, nitric oxide synthase inhibitor), 10(-6) mol/l 1H [1,2,4]oxadiazolo[4,3a]quinoxalin-1-one (ODQ, soluble guanylyl cyclase inhibitor) or 10(-5) mol/l zaprinast (cyclic GMP phosphodiesterase inhibitor). Ethanol (10 mmol/l) significantly decreased percent shortening from 10.0 +/- 0.9 to 6.0 +/- 0.2%. Similar decrements occurred in the maximum rate of shortening and relaxation. After L-NAME or ODQ, the decrements in percent shortening, maximum rate of shortening and relaxation caused by ethanol were not significant. After zaprinast, ethanol significantly decreased the maximum rate of shortening and relaxation and percent shortening to 4.3 +/- 0.5. Ethanol (10 mmol/l) significantly increased cyclic GMP from 403 +/- 121 to 529 +/- 128 fmol/10(5) myocytes. Both L-NAME and ODQ lowered cyclic GMP, and ethanol did not affect cyclic GMP after either. Zaprinast raised cyclic GMP, as did its combination with 10 mmol/l ethanol (653 +/- 120). Thus, ethanol both reduced myocyte function and increased cyclic GMP. Blocking nitric oxide production or guanylyl cyclase activity prevent these effects of ethanol, while blocking cyclic GMP degradation did not. This suggests that ethanol acts as a nitric oxide stimulator in ventricular myocytes leading to reduced function and increased cyclic GMP. PMID- 12566850 TI - Inhibition of prostaglandin biosynthesis in human endotoxin-stimulated peripheral blood monocytes: effects of caffeine. AB - There is an ongoing debate about possible advantages of the coadministration of caffeine with cyclooxygenase (COX) inhibitors in the treatment of pain. There are results suggesting interference by caffeine with COX expression and activity in rat immune cells. In the present study, we have used, therefore, human endotoxin stimulated monocytes to investigate a possible influence of caffeine on indometacin-induced inhibition of prostaglandin E(2) (PGE(2)) formation. Endotoxin caused a concentration- and time-dependent increase in immunoreactive PGE(2) that was dependent on CD14-mediated mechanisms. In order to investigate pharmacological inhibition of the COX activity, a submaximal concentration of 1 ng/ml endotoxin (4 h exposure time) was used. Indometacin caused a concentration dependent inhibition of PGE(2) with an apparent IC(50) of 8.9 +/- 1.4 x 10(-9) mol/l and an I(max) of 1 x 10(-7) mol/l. Caffeine (5 x 10(-6) to 1.5 x 10(-4) mol/l) on its own produced no statistically significant effect on endotoxin induced PGE(2) formation. In the presence of caffeine (5 x 10(-6) to 1.5 x 10(-4) mol/l), inhibition of PGE(2) biosynthesis by indometacin (1 x 10(-8) mol/l) was not significantly altered. These results show that, in human monocytes, caffeine, up to concentrations severalfold higher than those reached in patients, has no significant effect on endotoxin-induced PGE(2) formation nor on its inhibition by indometacin. PMID- 12566851 TI - Functional development of the rat urinary bladder after pre- or postpubertal castration. AB - Experiments were done to evaluate the role of sex hormones in the functional development of the rat urinary bladder. Rats were orchiectomized or ovariectomized at 30 (prepubertal) or 70 (postpubertal) days of age and bladders were removed 1 month later. An additional group of immature male and female rats was used in which the bladders were removed at 30 days of age. There were only minor differences in contractile responses of bladder strips from any group to electrical field stimulation, ATP, carbachol, or KCl compared to age-matched controls. There were no differences in responses of bladder strips from immature females or males to the adrenergic agonists, isoproterenol, norepinephrine, or methoxamine. Bladders from the pre- and postpubertally castrated rats and their controls relaxed fully in response to isoproterenol, but strips from prepubertally castrated rats relaxed significantly less in response to norepinephrine than those from other groups. Approximately one half of the bladder strips from prepubertally castrated rats failed to relax by at least 50% in response to norepinephrine; these same strips responded to methoxamine with exaggerated contractions. Our data indicate that the normal development of rat bladder alpha-adrenergic responsiveness is adversely altered by prepubertal castration. We postulate that this may result from an alteration in the relative expression of alpha(1)-adrenergic receptor subtypes. PMID- 12566852 TI - Mechanism of the protective effect of heteropolyoxotungstate against herpes simplex virus type 2. AB - The effects of heteropolyoxotungstate (K(7)[PTi(2)W(10)O(40)]. 6H(2)O; PM-19) on the replication of herpes simplex virus type 2 (HSV-2) were examined using a semiquantitative polymerase chain reaction of intracellular viral DNA established by us and also other methods. Vero cells were infected with HSV-2 strains: either the standard strain 169, or the acyclovir-resistant strain YS-4C-1. PM-19 was added at various stages during the replication of HSV-2. PM-19 strongly inhibited the synthesis of viral genomic DNA when it was added at the time of infection. The addition of PM-19 60-90 min after viral inoculation time-dependently decreased the antiviral activity and increased the relative yield of viral DNA, and the addition of PM-19 was completely ineffective at times later than 90 min. These results suggested that PM-19 inhibited viral penetration but did not affect the synthesis of viral DNA. Furthermore, PM-19 strongly inhibited a second round of infection. PMID- 12566853 TI - Paradoxical enhancement of bupivacaine anesthesia in mice by drugs that open sodium channels. AB - Sodium channel drugs were used to modulate the anesthetic effects of bupivacaine in mice. Anesthesia was measured following perisciatic injection of bupivacaine with vehicle or neurotoxin in the popliteal region. The site 1 Na(+) channel blocker tetrodotoxin alone was inactive, but increased the anesthetic effects of bupivacaine. We hypothesized that the site 2 and site 3 Na(+) channel openers veratridine and anemone toxin II (ATXII), respectively, would antagonize bupivacaine. Paradoxically, both drugs enhanced bupivacaine. In bupivacaine treated mice, a significant correlation was observed between limb weakness scores and paw withdrawal latencies. The correlation coefficients were higher when tetrodotoxin, veratridine, or ATXII was coadministered with bupivacaine. In conclusion, veratridine and ATXII may have increased the stimulus-dependent binding of bupivacaine to Na(+) channels, thereby increasing the anesthetic effects of bupivacaine. PMID- 12566854 TI - Role of protein kinases on acid-induced duodenal bicarbonate secretion in rats. AB - We examined the effect of several protein kinase inhibitors, such as staurosporine for protein kinase C (PKC), H-89 for protein kinase A (PKA) and genistein for tyrosine kinase (TK) on acid-induced duodenal bicarbonate secretion (DBS) in rats. HCO(-)(3) secretion was measured using the pH-stat method. Mucosal acidification was performed by perfusing the duodenal loop for 10 min with pH 2.2 HCl. Indomethacin, staurosporine and genistein were added to acidified saline and then perfused, respectively. In some cases, genistein and phorbol 12-myristate 13 acetate (PMA) were added to the luminal solution to examine the effect on basal duodenal HCO(-)(3) secretion. PGE(2) (PKA pathway) and PMA (PKC pathway) stimulate basal DBS. Indomethacin, H-89, staurosporine and genistein inhibit acid induced DBS, indicating involvement of the cyclooxygenase, PKA, PKC and TK pathways. PMID- 12566855 TI - Alterations of renal vasopressin V1A and V2 receptors in spontaneously hypertensive rats. AB - To elucidate the role of arginine vasopressin (AVP) in a hypertensive state, the characteristics of renal cortex V(1A) and medulla V(2) receptors in young spontaneously hypertensive rats (SHR) during the developmental phase of hypertension were compared with those of age-matched Wistar-Kyoto (WKY) rats using the radioligand receptor assay technique. The systolic blood pressure of 8 week-old SHR was statistically significantly higher than that of WKY rats (142 +/ 1 vs. 125 +/- 2 mm Hg). The plasma AVP levels were also significantly higher in SHR than in WKY rats (3.20 +/- 0.41 vs. 1.96 +/- 0.34 pg/ml). In SHR, the maximum capacity of (3)H-d(CH(2))(5)Tyr(Me)AVP binding to cortical V(1A) receptors (B(max)) was statistically significantly higher than that of WKY rats (39.7 +/- 2.7 vs. 22.4 +/- 0.9 fmol/mg protein). Furthermore, the B(max) values of (3)H-AVP binding to medullary V(2) receptors in SHR were also significantly higher than in WKY rats (40.2 +/- 1.9 vs. 28.3 +/- 1.3 fmol/mg protein). However, the apparent dissociation constant (K(d)) values of renal cortex V(1A) and medulla V(2) receptors in SHR and WKY rats were not significantly different. These results indicate that increased amounts of renal cortex V(1A) and medulla V(2) receptors in SHR play an important role in the pathophysiology of hypertension. PMID- 12566856 TI - Identification of a novel intercellular structure in late-stage differentiating lens cells. AB - This study describes a novel intercellular structure in the adult bovine lens. In differential interference contrast images, the structure has the shape of a thickened torus or 'bagel' of 3-9 micrometer diameter and is contributed equally by 2 adjacent fibre cells. Due to its shape and location reaching into 2 neighbouring cells, the novel structure was termed 'intercellular torus' or 'bagel'. Intercellular bagels are present in a subset of late-stage lens fibre cells of the intermediate cortex, a considerable time after the cytoplasmic organelles have been broken down and the pyknotic nuclear remnants have disappeared. They are not present in deeper fibres. Our experiments show that intercellular bagels do not stain positive for DNA or RNAs, but are rich in lipids. Preliminary data indicate that the intercellular bagels contain calcium, suggesting that they might act as a place of transient Ca(2+) storage or sequestration after the intracellular organelles, such as the endoplasmatic reticulum, nuclear envelope, Golgi apparatus and mitochondria have been eliminated from the lens fibres during terminal differentiation. PMID- 12566858 TI - Establishment of the mouse as a model animal for the study of diabetic cataracts. AB - The purpose of the present study was to investigate the suitability of using the mouse, a species known to have a low lens aldose reductase activity, as a model animal for studying the pathogenesis of diabetic cataract. Earlier studies with diabetic rats whose cataract development is much faster can only partially explain the etiology of cataracts in humans where lens aldose reductase is substantially low. CD-1 mice were injected intraperitoneally with streptozotocin according to Rossini's method. Blood glucose levels were estimated after 7 days, and animals having blood glucose between 300 and 400 mg/dl were selected for further experiments. Development of lenticular opacity was followed by examining the animals every 3-4 weeks by direct ophthalmoscopy, slitlamp examination and Scheimpflug photography. Additionally, the animals were sacrificed at appropriate intervals, eyes enucleated and subjected to morphological studies. The presence of refractive changes and early cataract in the diabetic mice was initially ascertained by the distorted appearance of the grid pattern when seen through the isolated lenses. Early cataracts were visible on slitlamp examination and by ophthalmoscopy as early as 3-4 weeks after the establishment of diabetes. Advanced opacity was clearly documentable by photography after 5-6 months. Similar to that in other species, a single layer of anterior epithelial cells abutting the anterior capsule was seen in the histological sections of normal mouse lenses. On the contrary, the epithelium in the diabetic lens was multilayered, and numerous nucleated cells were visible in the superficial anterior cortex. These studies therefore suggest that further studies with mice may throw additional light on the contribution of diabetes in the pathogenesis of cataracts in low lens aldose reductase models. PMID- 12566857 TI - Identification of a vimentin-reactive Peptide associated with ocular lens membranes as cytokeratin. AB - We wished to identify a 44-kD peptide isolated from lens membrane fractions which has apparent immunoreactivity to antivimentin and to antiphakinin. Urea-soluble proteins from rat lens membrane fractions were separated by two-dimensional electrophoresis, transferred to polyvinylidene fluoride membranes and probed with monoclonal antibodies to vimentin or cytokeratins, or a polyclonal antibody to phakinin. The monoclonal antibody to vimentin recognized the expected protein (M(r) = 56 kD, pI = 5.1) and several smaller and more acidic peptides including a 44-kD spot with a pI of 4.9. A similar pattern of immunoreactivity was found with commercially available purified vimentin. The polyclonal antibody to phakinin recognized the expected protein (M(r) = 52 kD, pI = 5.0) and several smaller and more acidic peptides, including a 44-kD spot with a pI of 4.9. Matrix-assisted laser desorption ionization mass spectroscopy analysis of the tryptic digest of the spot corresponding to 44 kD and pI of 4.9 identified neither vimentin nor phakinin but did identify peptides derived from ovalbumin (added to samples as an internal standard) and cytokeratin 1. Antibodies which recognize cytokeratin 1 reacted weakly with a 44-kD peptide which comigrated with the 44-kD vimentin immunoreactive peptide in adjacent lanes of single-dimension SDS polyacrylamide gels. The 44-kD acidic (pI = 4.9) peptide which reacts with a monoclonal antibody to vimentin may be derived from cytokeratin 1. PMID- 12566859 TI - Glare disability and driving safety. AB - PURPOSE: Increasing investigation of the visual elements of safe driving environments may be of great benefit to society. Visual disability appears to be only one of many visual factors related to traffic accidents. The purpose of this article was to examine the type of visual impairment mediated by the increased glare sensitivity in adult drivers using the original halometer glare test. METHODS: In this article, the visual sensory, cognitive and motor functions relevant to driving, their measurement, the epidemiology and prevention of age associated functional impairments and the relationship of functional impairments to both self-reported driving and the imposition of legal restrictions are reviewed. RESULTS: The problem of night and tunnel driving is the most urgent in relation to the effects of glare from vehicle headlights on motion perception of drivers. The reduced mesopic vision and increased sensitivity to glare are accompanied by an increased risk of nighttime accidents. Elderly drivers and patients with beginning cataract cannot sufficiently fulfill the criteria for night driving ability because of contrast and glare sensitivity. It is indispensable for the parameters mentioned to be carefully measured and for drivers to be informed that night driving ability may be impaired, even if visual acuity is sufficient. CONCLUSIONS: It would be advisable for traffic safety if simple tests for contrast and glare sensitivity were implemented for vehicles and/or were regularly added to the requirements for a driver's licence, at least for older drivers. The age, functional status and test result limits should be defined to avoid a risk factor in traffic. PMID- 12566860 TI - Augmentation of galactose-specific glycoproteins during corneal epithelial migration in organ culture. AB - The purpose of the present study was to identify galactose-specific glycoproteins that are synthesized in higher amounts or downregulated during the migratory process of corneal epithelium. Trichloroacetic-acid-precipitable proteins showed an increased incorporation of (3)H-galactose with time in migrating corneal epithelium in organ culture. Galactose oxidase treatment of immobilized 3-[(3 cholamidopropyl)dimethylamminio]-1-propanesulfonate-extractable glycoproteins of migrating and nonmigrating corneal epithelia resolved on sodium dodecylsulfate polyacrylamide gel electrophoresis was carried out. Two prominent terminally galactosylated protein bands (180 and 98 kD) were seen in the migrating epithelium but were absent in the nonmigrating epithelium. We conclude that during the migratory process of the corneal epithelium there is an increased utilization of galactose, and galactose-specific glycoproteins are synthesized in higher amounts. PMID- 12566861 TI - Experimental dispase-induced retinopathy causes up-regulation of P2Y receptor mediated calcium responses in Muller glial cells. AB - During proliferative vitreoretinopathy (PVR) Muller glial cells show an up regulation of their responsiveness to extracellular adenosine 5'-triphosphate (ATP). In the present study, we investigated if such a glial cell response is also a feature for other retinopathies besides PVR. To this aim, the proteolytic enzyme, dispase (0.1 U), was injected into the vitreous of rabbit eyes. After 3 weeks, a distinct retinopathy had developed which showed no signs of PVR. The retinopathy was characterized by strong alterations of the retinal vasculature in the medullary rays, by photoreceptor degeneration, retinal atrophy, and activation of microglial cells. Muller cells became reactive, as indicated by up regulation of glial fibrillary acidic protein immunoreactivity and by hypertrophy involving subretinal fibrosis. Muller cell reactivity was also evidenced electrophysiologically by a down-regulation of their inwardly rectifying potassium currents and by an up-regulation of their responsiveness to extracellular ATP. Significantly more Muller cells from dispase-treated eyes showed ATP-evoked calcium (83%) and current responses (69%) when compared with cells from control eyes (13 and 9%, respectively). The results indicate that increased responsiveness to extracellular ATP may be a more general feature of Muller cell gliosis, and is also observed in retinopathies besides PVR. PMID- 12566862 TI - Transcription of a single mannose receptor gene by macrophage and retinal pigment epithelium. AB - To determine if the macrophage mannose receptor transcript is present in mouse, rat, pig, and human retinal pigment epithelium (RPE), primary cultures and/or freshly dissected retinal pigment epithelium from four different species were used to isolate total RNA. RT-PCR was used to amplify segments of the macrophage mannose receptor from each sample. Amplified products were sequenced and compared with known sequences of the macrophage mannose receptor. Macrophage mannose receptor transcripts were identified in all RPE samples. Comparison between sequences identified in RPE with macrophage sequences from the same species revealed 100% identity. Sequence homology between the different species was 74% or greater. These data are consistent with the transcription of a single mannose receptor gene by these two phagocytic cell types. PMID- 12566863 TI - Survival of allografted pancreatic islets in the subretinal space in rats. AB - The purpose of this study was to determine whether the subretinal space would provide immune privilege for pancreatic islet transplantation. Islets from outbred Sprague Dawley (SD) rats were isolated by collagenase digestion, and about 200 handpicked islets were transplanted into the subretinal space of SD rats. Similar grafts were transplanted into the subconjunctival space of SD rats as controls. Transplanted eyes were enucleated after 2 to 60 days, fixed and embedded in paraffin for immunoperoxidase staining of insulin, glucagon, and CD8+ lymphocytes. Clinical examination of rat eyes revealed minimal or no inflammation in the anterior chamber or vitreous at any time point. Fifteen of 19 subretinal allografts survived up to 60 days. Few CD8+ lymphocytes were present in the subretinal grafts and the endocrine cells stained intensely for insulin and glucagon at all time points. In contrast, CD8+ lymphocytes were present in subconjunctival grafts in rats by day 14 and all grafts were destroyed by day 21. These results suggest that the subretinal space provides immune privilege for islet allotransplantation by preventing massive lymphocyte infiltration. PMID- 12566864 TI - Effect of a yellow filter on mesopic contrast perception and differential light sensitivity in the visual field. AB - PURPOSE: Previous studies on the effects of yellow filters in the visual performance have been carried out, presenting discrepancies among the different investigations. Our study reports the influence of the yellow filter without residual reflections on contrast perception and differential light sensitivity (DLS) under mesopic conditions. METHODS: Contrast sensitivity, low contrast visual acuity and luminance thresholds in the central visual field (30 degrees ) were measured in a group of 38 adult subjects with and without a coated yellow lens (482-nm cut-off) under mesopic conditions. RESULTS: The contrast sensitivity mean was significantly better with the filter at low- and middle-range spatial frequencies (1.5 cycles/degree p = 0.002 and 6 cycles/degree p = 0.02). At 5% contrast, the mesopic low contrast visual acuity improved significantly (p = 0.004) when interposing the yellow filter. The increase in DLS was statistically significant [e.g. all visual fields p = 0.0003 and macular visual field (9.5 degrees ) p = 0.00001] through the yellow filter on the different established areas of the visual field (30 degrees ). CONCLUSIONS: The yellow filter without residual reflections had a positive influence on the brightness and contrast perception of healthy subjects, for which its use would be beneficial under mesopic conditions. PMID- 12566865 TI - Treatment of rubeosis iridis with photodynamic therapy with verteporfin--A new therapeutic and prophylactic option for patients with the risk of neovascular glaucoma? AB - Patients with ischaemic retinopathy who show iris neovascularization despite panretinal laser photocoagulation (PRP) very often develop a neovascular glaucoma. Photodynamic therapy (PDT) has been shown to occlude neovascularization without damage to physiologic vessels or adjacent tissue in the treatment of choroidal neovascularization (CNV) and might also be of value for patients with neovascular glaucoma who did not benefit from the PRP. First results of a monocentre, open label, intra-individual controlled, pilot phase I/II, dose finding study demonstrate that PDT with verteporfin is capable of occluding neovascular vessels for a defined period of time without damaging adjacent tissue or physiologic iris vessels. Whether this vessel occlusion will have an impact on the progression of rubeosis or neovascular glaucoma will be the subject of further investigation. PMID- 12566866 TI - Clinical effects of the Nd:YAG laser operating in the photodisruptive and thermal modes. A review. AB - The clinical effects of the Nd:YAG laser operating in both the photodisruptive (Q switched) and thermal (free-running, cw) modes are discussed, and their clinical applications investigated. Moreover, the physical background of the working modes is explored. When working in the photodisruptive and fundamental (TEM(00)) modes, it is possible to carry out delicate clinical tasks with minimally invasive effects. When the laser is being used in the multimode regime, tasks that are highly resistant to photodisruptive laser radiation can be undertaken. In the thermal mode, photocoagulation can be performed. Nd:YAG laser light (1,064 nm) has high optical tissue penetration and good hemostatic properties, particularly when it is being operated in the frequency-doubled mode (KTP laser). PMID- 12566867 TI - Low specificity of scanning laser polarimetry. AB - PURPOSE: Scanning laser polarimetry is used to assess the peripapillary retinal nerve fiber layer. This study evaluates scanning laser polarimetry in normal subjects. SUBJECTS AND METHODS: 28 eyes of 28 normal subjects (age range 21-48 years, mean age 36.5 years) had scanning laser polarimetry (NFA-GDx, Laser Diagnostic Technologies, San Diego, Calif., USA). All subjects had normal eye examinations, refractive errors with spherical equivalents <5 D and astigmatism <2 D, normal intraocular pressures, no history of diseases affecting the visual field or nerve fiber layer, and normal white-on-white automated perimetry (Octopus 101, program G2). The mean image of three good-quality measurements with the scanning laser polarimeter was included. RESULTS: 36% of the normal subjects (10/28) had at least one NFA parameter outside normal (p < 0.05). The parameters most frequently found to be outside normal were symmetry (21%, 6/28) and superior ratio (17%, 5/28). CONCLUSIONS: Scanning laser polarimetry is often used for early detection of glaucomatous damage in glaucoma suspects. This study found a specificity of only 64% and indicates that the actual selection of parameters is not adequate. Especially the parameters symmetry and superior ratio should be reevaluated in order to achieve a higher specificity. PMID- 12566868 TI - Ultrasound biomicroscopy of eyelid lesions. AB - This study compared ultrasound biomicroscopy with histopathology in discerning depth of eyelid lesions and tissue characteristics. Ultrasound biomicroscopy (50 MHz) was used to analyze lesion depth and tissue characteristics of eight patients with eyelid lesions referred to a university-based referral practice prior to excisional biopsy. This data was then compared to histopathologic findings. Lesion depth measured by ultrasound biomicroscopy was correlated with histopathologic depth with a linear regression. Ultrasound biomicroscopy correlated highly to histopathology with regards to tissue characteristics and lesion depth (R(2) = 0.87). Ultrasound biomicroscopy accurately measures depth of eyelid lesions and accurately determines tissue characteristics. PMID- 12566869 TI - Trabeculectomy with mitomycin C versus trabeculectomy alone for juvenile primary open-angle glaucoma. AB - PURPOSE: We compare the intermediate-term outcome of initial trabeculectomy with adjunctive mitomycin C use versus initial trabeculectomy alone for juvenile primary open-angle glaucoma. METHODS: This retrospective consecutive analysis included 44 eyes from 36 patients with juvenile primary-open angle glaucoma, all of whom underwent either initial trabeculectomy with adjunctive mitomycin C use (15 eyes) or initial trabeculectomy alone without mitomycin C use (29 eyes). We compared the success rate and complications between the two groups in a three year follow-up period following surgery. RESULTS: Three years subsequent to surgery, the cumulative success probability was 73% for the mitomycin C group and 68% for the control group, there being no real difference between the two groups (p = 0.89). A greater incidence of hypotony maculopathy was found amongst the mitomycin C group than was the case for the control group (20 versus 0%, respectively, p = 0.034). A lower intraocular pressure amongst the mitomycin C group was noted as compared with the control group (10.8 +/- 3.0 versus 13.3 +/- 3.8 mm Hg, respectively, p = 0.017) amongst the successfully treated patients. CONCLUSIONS: Despite the lower intraocular pressure level for the successfully treated patients from the trabeculectomy with mitomycin C group, and a greater incidence of resultant hypotony maculopathy for this group as compared with the trabeculectomy alone group, there appeared to be no significant difference in the cumulative success probability for this group as compared with the trabeculectomy alone group. Therefore, we caution against the use of an initial trabeculectomy with mitomycin C for juvenile primary open-angle glaucoma. PMID- 12566870 TI - Comparison of efficacy and tolerability between two gel-forming timolol maleate ophthalmic solutions in patients with glaucoma or ocular hypertension. AB - PURPOSE: To compare two gel-forming timolol maleate ophthalmic solutions, Timoptol XE and Lizmon TG, in regard to efficacy and tolerability in patients with glaucoma or ocular hypertension by means of a patient-masked prospective randomized crossover study. SUBJECTS AND METHODS: A total of 37 patients with glaucoma or ocular hypertension under treatment with antiglaucoma ophthalmic solutions including 0.5% twice-daily timolol maleate participated in this study. Only timolol maleate was withdrawn and either Timoptol XE or Lizmon TG was randomly allocated. After instillation for 1 month, the other ophthalmic solution was subsequently instilled for another month. Routine ophthalmic examination including slit-lamp examination and intraocular pressure (IOP) monitoring was conducted before instillation of gel-forming ophthalmic solutions and just after completing the instillation of each ophthalmic solution. Patient questionnaire surveys were also performed just after completing the instillation of each ophthalmic solution. RESULTS: Mean IOP, just before the withdrawal of timolol ophthalmic solution, was 16.4 +/- 2.9 mm Hg. At the end of Timoptol XE or Lizmon TG instillation, mean IOPs were 16.3 +/- 2.5 or 16.3 +/- 3.0 mm Hg, respectively. The results of the questionnaire survey showed no significant difference between Timoptol XE and Lizmon TG in regard to all survey items. Twenty-nine patients (78.4%) preferred to use gel-forming timolol solutions rather than twice-daily timolol ophthalmic solution. The presence of concurrently used ophthalmic solutions did not effect the incidences of subjective symptoms. The incidences of objective adverse effects were not significantly different between two gel forming timolol ophthalmic solutions. CONCLUSION: Both gel-forming timolol ophthalmic solutions could be good choices for glaucoma treatment. PMID- 12566871 TI - Ocular perfusion pressure and visual field indice modifications induced by alpha agonist compound (clonidine 0.125%, apraclonidine 1.0% and brimonidine 0.2%) topical administration. An acute study on primary open-angle glaucoma patients. AB - OBJECTIVE: The aim of this study was to verify the acute effects of alpha adrenoreceptor agonist eyedrop administration on visual field parameters and ocular perfusion pressure (OPP) in patients affected by primary open-angle glaucoma. METHODS: A prospective, randomized double-blind study was carried out. Sixty-four glaucomatous subjects were enrolled in the clinical trial and subsequently separated into four study groups, of 16 patients each, to compare the systemic and ocular effects of placebo, clonidine 0.125%, apraclonidine 1.0% and brimonidine 0.2%. RESULTS: No significant variations in the ocular or systemic parameters were observed after placebo administration. All the alpha adrenoreceptor agonist compounds induced a significant reduction of the intraocular pressure. Clonidine induced significant modifications of mean blood pressure, OPP and visual field indices. The acute administration of apraclonidine did not affect both mean blood pressure and OPP, but a worsening of the visual field was nevertheless recorded. The analyzed parameters did not significantly vary after brimonidine instillation. CONCLUSIONS: The present findings demonstrate that the lack of effects on the blood flow and the absence of vasomotor activity at the level of the posterior pole exhibited by brimonidine is related to its alpha-2 selectivity, as appears by comparing this compound with the other alpha-agonists available for the management of glaucoma. PMID- 12566872 TI - Detection of CMV DNA in the aqueous humor of AIDS patients with CMV retinitis by AMPLICOR CMV test. AB - The present study was performed to detect the cytomegalovirus (CMV) DNA in the aqueous humor from the eyes of acquired immunodeficiency syndrome (AIDS) patients with CMV retinitis. Detection of CMV DNA in the aqueous humor in the eyes with active CMV retinitis was compared with detection of CMV DNA in inactive retinitis. CMV DNA in the aqueous humor was evaluated before and after treatment with intravitreal injection of ganciclovir. CMV DNA in the aqueous humor was measured by AMPLICOR CMV test. Forty-two eyes of 35 AIDS patients were diagnosed ophthalmoscopically as having CMV retinitis that was subclassified as either active or inactive. The active and inactive CMV retinitis cases were distinguished based on clinical evaluations and fundus photographs. The results showed that 37 of the 42 eyes (88.1%) were positive for CMV DNA prior to treatment, while in 29 of these 37 eyes (78.4%), the aqueous humor became CMV DNA negative after the treatment. Successful treatment with the intravitreal injection of ganciclovir was associated with a reduction in the detection of CMV DNA in the aqueous humor. CMV DNA was not detected in the aqueous humor of patients with quies cent CMV retinitis. In conclusion, the AMPLICOR CMV test was found to be a reliable tool for differentiating active and inactive CMV retinitis, and is useful for helping to select the optimal treatment regimen. The intravitreal injection of ganciclovir is highly effective in reducing detectable CMV DNA in the aqueous humor. PMID- 12566873 TI - Aqueous flare and macular edema in eyes with diabetic retinopathy. AB - BACKGROUND: The breakdown of the blood-retina barrier in diabetic patients is correlated with a dysfunction of the blood-aqueous barrier. In our study, we wanted to investigate the effect of grid photocoagulation in diabetic macular edema on the aqueous flare. METHODS: Modified grid pattern photocoagulation was performed on 20 patients with diabetic macular edema. The aqueous flare intensity was measured by the laser flare cell meter before and 4 months after laser treatment. RESULTS: The mean flare value was 9.8 +/- 4.7 photons/ms before laser treatment. After grid photocoagulation, the flare value was 7.1 +/- 3.6 photons/ms. Flare values were significantly decreased after laser treatment. CONCLUSION: Our results suggest a correlation between the effect of grid photocoagulation on the blood-retina barrier and the permeability of the blood aqueous barrier. The laser flare intensity seems to be a quantitative indicator for the diabetic dysfunction of the blood-retina barrier. PMID- 12566874 TI - Elevated adrenomedullin in the vitreous of patients with diabetic retinopathy. AB - Adrenomedullin (AM) is a multifunctional peptide with various physiological actions, including vasodilatation, a defense mechanism against microorganisms, the regulation of growth and the regulation of insulin and glucose. In this study, we measured the vitreous AM levels in patients with diabetes mellitus to determine its potential involvement in the pathogenesis of diabetic retinopathy (DR). We used an immunoradiometric assay to measure the vitreous AM concentrations in a total of 28 eyes: 13 with DR and 15 with macular holes (15 men and 13 women, 62.9 +/- 10.4 years old). The AM levels in the vitreous fluid of patients with DR (22.9 +/- 7.9 fmol/ml) were found to be significantly higher than the corresponding AM levels in patients with macular holes (4.7 +/- 1.1 fmol/ml) (p < 0.05). These results indicate that the increase in the vitreous AM is related to DR. PMID- 12566875 TI - Early and late assessment of internal drainage of chronic dacryocystitis. AB - To evaluate the efficacy of internal drainage of a lacrimal abscess, we treated 10 patients with pyocele of the lacrimal sac, who did not want to undergo operative intervention. A temporary nasolacrimal stent was placed in the nasolacrimal duct, and the pussy material was collected for microbiological examination to adjust the preliminary antibiotic treatment which was given systemically and locally. The stents were removed after the infection had subsided clinically. Three years later, there was a recurrence of the infection in only 1 patient. This internal drainage of the lacrimal sac, combined with systemic and local antibiotics, was successful in 90% of the patients. PMID- 12566876 TI - Holographic interferometry of excimer-laser-ablated bovine eyes: first results. AB - PURPOSE: To investigate the effects of areal corneal ablations of different depth on the biomechanical properties of the cornea. METHOD: The qualitative change of the holographic interferometric fringe pattern of enucleated bovine corneas due to areal mechanical ablations (group A) of 35% and 70% depth of the central cornea and 15, 30, and 50% excimer laser ablations in each eye (group B) was investigated. RESULTS: Ablations of 50% (group B) and 70% (group A) showed a significant variation of the fringe pattern as an effect of the corneal weakening. 15% excimer laser ablations of 5 mm diameter did not result in noticeable changes of the fringe pattern. Half of the 30% ablated corneas in group B showed newly appearing fringes within the area of ablation, indicating a local reduction of the corneal stiffness after ablation. CONCLUSION: The thickness of the remaining tissue is very important for the biomechanical behavior of the cornea after areal ablations. There are interindividual differences regarding the change of material properties due to excimer laser ablation of 30% depth of ablation. PMID- 12566877 TI - Biomolecular markers of malignancy in human uveal melanoma: the role of the cadherin-catenin complex and gene expression profiling. AB - In recent years there has been a trend towards conservative management of uveal melanoma (UM), aimed at preserving the eye and vision. Despite improvements with this approach, recurrent tumour and metastatic disease still occur, and the management remains problematic. As a result of these limitations, there is interest in gaining a greater understanding of molecular changes associated with aggressive disease patterns in UM. This might result in new, more effective and less toxic therapies as well as provide prognostic information for defining subgroups of patients with a less favourable prognosis as potential candidates for adjuvant therapies. Accumulating evidence over the past decade suggests that disturbance in the cadherin-catenin adhesion complex is critical in the process leading to invasion and metastasis of many cancers. The recent advent of DNA micro-array technology now offers an unprecedented ability to study these molecules and others associated with malignant transformation. In this mini review, the aspects of tumour progression in which cadherin-catenin may be involved are dealt with along with the potential application of DNA micro-array technology to the problem in UM. PMID- 12566878 TI - Orbital involvement in multiple myeloma: first sign of insufficient chemotherapy. AB - Orbital involvement in multiple myeloma is unusual. We describe the case of a 60 year-old male who presented with left proptosis, reduced visual acuity, diplopia and progressing signs of globe indentation 2 months after chemotherapy for multiple myeloma. MRI showed a well-defined tumor filling the mediobasal part of the left orbit. Incisional biopsy and reduction of tumor mass were performed using an anterior transconjunctival orbitotomy. Histopathologic findings and further systemic examination confirmed the reactivation of the multiple myeloma. Proptosis, intraocular pressure and visual acuity improved following external beam radiation therapy of the left orbit and repeated systemic chemotherapy. Orbital involvement in multiple myeloma was the only sign of insufficient chemotherapy. PMID- 12566879 TI - Ischemic optic neuropathy associated with subarachnoid hemorrhage after rupture of anterior communicating artery aneurysm. AB - Two clinical cases in which ischemic optic neuropathy (ION) occurred after subarachnoid hemorrhage (SAH) are reported. Hemorrhage in the proximity of the optic chiasm was confirmed in 2 cases following rupture of an anterior communicating artery aneurysm. Optic disk atrophy with excavation and permanent visual field defect (altitudinal superior hemianopia) occurred in both cases. ION seems to occur in association with the optic nerve coincidental with the hyperdensity side of SAH on head CT scan. The incidence of ION appears to be attributable to an insufficient blood supply to arteries distributed in the posterior part of the optic nerve as a result of SAH. PMID- 12566880 TI - Applications of the neodymium:YAG laser in plastic surgery of the face and lacrimal surgery. Wound repair. A review. AB - The physical and clinical features of a number of laser energy sources presently being used in plastic and endonasal surgery, such as the Nd:YAG, KTP, Ho:YAG, Er:YAG and CO(2) lasers, are emphasized. Particular attention is paid to the Nd:YAG laser and to an Nd:YAG laser-powered quartz laser scalpel. PMID- 12566881 TI - Human cerebellar activation in relation to saccadic eye movements: a functional magnetic resonance imaging study. AB - PURPOSE: The functional organization of the human cerebellum involved in saccadic eye movements was investigated using functional magnetic resonance imaging (fMRI). METHODS: The subjects were 7 normal volunteers aged 18-34 years. Visual stimuli were back-projected onto a screen placed at the subjects' feet. The stimulation period of 30 s consisted of a saccade target jumping back and forth horizontally by 20 degrees once per second. The control period of 30 s consisted of a fixed target. The stimulation and control periods were alternated 10 times during the presentation. Functional images were collected with a 1.5-tesla clinical MRI scanner. The significance of activation was determined by Statistical Parametric Mapping (SPM 99) at a threshold of p < 0.001 (uncorrected), and significantly activated areas were superimposed on the T(1) weighted images. RESULTS: Significantly activated areas related to visually guided saccades were observed in the cerebellar vermis (declive and folium), in the bilateral cerebellar hemispheres (mainly the superior semilunar lobule) of the cerebellum, in the frontal eye field, in the supplementary eye field and in parts of the parietal lobule of the cerebrum. CONCLUSION: Our results suggest that the cerebellar posterior vermis and bilateral hemispheres are related to saccades in humans. These results are consistent with neurophysiological data obtained in primates. PMID- 12566882 TI - Normal isopter position in the peripheral visual field in goldmann kinetic perimetry. AB - PURPOSE: Often in young patients the question arises if a disease state has contracted their peripheral boundary of the visual field. Since the 'hill of vision' is steeper in the periphery, kinetic perimetry is more sensitive in detecting peripheral visual field abnormalities than static perimetry. In order to be able with kinetic perimetry to detect mild peripheral visual field constrictions, we determined the normal position of 4 isopters in the peripheral visual field. SUBJECTS AND METHODS: Intraindividual sensitivity variations of the isopters in the peripheral visual field were determined in 3 normal subjects by one perimetrist. Then, in 22 subjects (19-42 years old) the position of the isopters V4e, III4e, I4e and I3e has been determined by one perimetrist. Visual fields were registered using a Goldmann kinetic perimeter. RESULTS: The intraindividual sensitivity variations (measured as 1 standard deviation) for the isopter V4e was 0.98 degrees and for the isopter I4e 1.13 degrees. To visualize the normal isopter positions in the peripheral visual field, the average positions +/- 2 standard deviations were plotted for the isopters V4e, III4e, I4e and I3e. CONCLUSION: In this study, we determined the normal position of 4 isopters frequently used to plot the peripheral visual field. The plots have been printed in a scale of 1:4 in order to facilitate its use. Photocopying the figures on transparencies by scaling them with a factor 4 will allow to superpose the transparencies on individual Goldmann kinetic visual fields in order to easily determine whether the individual isopters are inside or outside the normal range. PMID- 12566883 TI - Variance between program versions in measuring optic nerve fiber layer thickness using optical coherence tomography. AB - PURPOSE: To compare nerve fiber layer thickness (NFLT) in patients with glaucoma measured by two versions of an analysis program utilizing optical coherence tomography (OCT) with reference to the cup-to-disk (C/D) ratio and the mean deviation (MD) score of a static visual field test. METHODS: Twenty-four eyes from 24 glaucoma patients were enrolled. The NFLT was measured by OCT along a circle around the disk with the circle diameter being 1.5 times that of the disk itself. The NFLTs evaluated by the two versions A4.01 and A6.01 were compared with respect to the C/D ratio and MD score. RESULTS: The mean NFLTs evaluated by version A6.01 were 61.2% greater than those evaluated by version A4.01 (p < 0.001). The NFLTs in all quadrants evaluated by A6.01 were also greater than those evaluated by A4.01. Both A6.01 and A4.01 showed a significant correlation with the C/D ratio and MD score. Although A6.01 showed a better correlation with the C/D ratio than A4.01, both A4.01 and A6.01 showed similarly good correlations with MD. CONCLUSION: The NFLTs evaluated by the two versions differed significantly with regard to some parameters. These results should be taken into account in evaluating NFLTs when utilizing OCT. PMID- 12566884 TI - Pattern visual evoked potentials in traumatic optic neuropathy. AB - To evaluate the characteristics of pattern visual evoked potentials (PVEPs) in traumatic optic neuropathy (TON) compared with optic neuritis (ON), we retrospectively studied 47 patients with TON and 64 patients with unilateral ON. PVEPs elicited by the stimulation with 3 reversals/s were recorded in all patients. A PVEP could be recorded in 29.8% of the TON patients and 49.6% of the ON patients at their first visit. In the 14 patients with TON and recordable PVEPs, the mean amplitude ratio (affected/fellow eye) was 0.29 +/- 0.03 (SE) and the mean delay of the peak latency was 17.9 +/- 2.9 ms. In the 30 patients with ON and recordable PVEPs, the mean amplitude ratio was 0.48 +/- 0.04 (SE) and the mean delay of the peak latency was 23.5 +/- 2.5 ms. The amplitude ratio was significantly lower in patients with TON than in those with ON. PMID- 12566885 TI - Subclinical experimental optic neuropathy after accelerated proton beam irradiation. AB - BACKGROUND: Accelerated proton beam irradiation has been used for several years to treat intraocular tumors. The pathophysiology of proton-beam-induced retinopathy and neuropathy has not been characterized to date. The present study investigates the early effects of irradiation with an accelerated proton beam on the optic nerve of the rabbit. MATERIALS AND METHODS: The optic nerve head of Albino New Zealand rabbits (n = 14) was irradiated with a narrow beam of accelerated protons using the total dose of 60 and 43 Gy, respectively. This dose was split up into 4 equal sessions taking place on 4 consecutive days. Ophthalmoscopic examination was performed regularly, and the rabbits were sacrificed at 1, 3 and 8 months after irradiation. The eyes were enucleated and processed for light and electron microscopy. RESULTS: Despite the absence of ophthalmoscopically detectable optic neuropathy in all 14 rabbits irrespective of the dose of irradiation, light and electron microscopy of the optic nerve showed a glial and fibrotic perivascular scar made up predominantly of altered astrocytes. This scar formation was seen as early as 1 month after irradiation and was at times accompanied by infiltration with inflammatory cells perivascularly both outside and within the optic nerve. In contrast to the astrocytes, oligodendrocytes did not show degenerative cellular alterations. During the 8-month follow-up, no signs of vascular occlusion were found. CONCLUSIONS: The observed lesions in the glial tissues with consecutive fibrosis appear to stem from a direct effect of irradiation. This may represent the initial mechanism of early subclinical irradiation-induced damage to the optic nerve before vascular occlusions may occur at a later stage, which may lead to more severe damage. PMID- 12566886 TI - Occlusion therapy for the treatment of amblyopia: letting the parents decide. AB - BACKGROUND: Compliance with prescribed occlusion therapy is a significant problem in the treatment of amblyopia. Parental preference for a particular type of occlusion treatment has not been previously addressed. Unless parental views are taken into account when planning therapy, compliance may be poor and treatment may fail. SUBJECTS AND METHODS: Parents of children with strabismic and/or anisometropic amblyopia who were due to start or had already had occlusion treatment were included in this study. Group A comprised parents of children who had no previous experience of occlusion, and group B comprised parents of children who had previous experience of occlusion therapy. Parental preferences regarding occlusion therapy were investigated by way of a questionnaire, in which they were asked whether they would prefer part-day/full-week occlusion or all day/part-week occlusion so that the total number of hours of occlusion per week was the same. RESULTS: One hundred parents completed the questionnaire, 47 from group A and 53 from group B. A significant number of parents in group A (95.3%) who had no previous experience with occlusion preferred part-day/full-week occlusion (p < 0.001), whereas there was no such preference among parents who had experience with occlusion, that is only 54.3% of parents in group B showed a preference for part-day/full-week occlusion (p = 0.1). The reasons given by the parents for their preferences varied but were in keeping with their lifestyles so that the type of occlusion regimen chosen by the parents was in accordance with the reasons given for their choice. CONCLUSION: Parental preferences should be considered when occlusion therapy is planned if compliance is to be improved. For maintenance occlusion, it would be reasonable to prescribe the number of hours of occlusion required per week and allow parents the responsibility to implement the pattern of occlusion according to their circumstances--the implied restriction ensuring that the number of hours of occlusion per week is met. Although parents appeared to have preconceived ideas before the commencement of treatment, their preferences changed once treatment had begun. PMID- 12566887 TI - Does photorefractive keratectomy alter the effect of topical medications? AB - INTRODUCTION: The influence of photorefractive keratectomy (PRK) on drug penetration into the eye is not yet established, as this procedure may alter the barrier function of the cornea as well as the blood ocular barrier. Pupillary response to topical tropicamide depends on its penetration into the anterior chamber. The purpose of this study was to examine the influence of PRK on pupillary response to topical tropicamide, and hence to evaluate whether PRK alters drug penetration into the eye. METHODS: Pupillary diameter was measured using infrared pupillometer at 0, 15 and 30 min after instillation of 0.1% tropicamide to both eyes of 28 patients 4 weeks after PRK in one eye. The untreated eye served as control. RESULTS: Pupillary diameter before and 30 min after dilation was 7.4 +/- 1.2 and 8.0 +/- 1.0 mm in the treated eyes and 7.5 +/- 1.2 and 8.1 +/- 1.0 mm in the untreated eyes, respectively (p = 0.45). Percentage of pupillary dilation (delta pupillary diameter/pupillary diameter at 0 min x 100) was also similar in treated and untreated eyes for the entire study group (12.5 and 10.5%, respectively, p = 0.17). A trend for greater dilation was seen in patients with myopia above 6 diopters (13.7 and 10.5% in treated and untreated eyes, respectively, p = 0.11) and cornea thinner than 540 microm (14.2 and 10.1% in treated and untreated eyes, respectively, p = 0.1). DISCUSSION: Our results confirm animal and human studies that found restoration of the ocular drug barrier at 4 weeks following PRK. The trend for greater penetration in patients with high myopia and thin corneas warrants further study on a larger cohort of this subgroup. PMID- 12566888 TI - Ocular blood flow in healthy and primary open-angle glaucomatous eyes. AB - PURPOSE: To evaluate blood flow parameters in the ophthalmic artery, in the posterior ciliary arteries and in the central retinal artery in eyes with primary open-angle glaucoma. METHODS: Hemodynamic parameters were evaluated by color Doppler imaging in 48 patients with primary open-angle glaucoma and 42 age matched normal control subjects. Peak systolic velocity (Pv), end-diastolic velocity (Ev), and resistive index (RI) were calculated in the central retinal artery, short posterior ciliary arteries and the ophthalmic artery. RESULTS: The analysis of the ocular blood flow velocities in the central retinal artery, short posterior ciliary arteries and the ophthalmic artery revealed a significant reduction in peak systolic velocity and end-diastolic velocity with an increase in resistive index in primary open-angle glaucoma compared with controls (p < 0.05). CONCLUSION: Our data indicate primary open-angle glaucoma to be associated with a decreased flow velocity and increased resistive index in the ocular vasculature. PMID- 12566889 TI - Effects of amniotic membrane transplantation and mitomycin C on wound healing in experimental glaucoma surgery. AB - PURPOSE: To evaluate the effects of mitomycin C and amniotic membrane transplantation on the modulation of wound healing after glaucoma filtration surgery. METHODS: Trabeculectomy was performed in 3 groups of 10 rabbit eyes each. Sponges soaked in mitomycin C were applied to the first group, and amniotic membranes were transplanted around the scleral flap in the second group; the third group was the control. The arithmetic means of fibroblast and macrophage numbers per square centimetre were calculated. RESULTS: The mean numbers of fibroblasts and macrophages were decreased significantly in the mitomycin C group, compared with the controls (p < 0.0001). Cell counts in the amniotic membrane group were also lower than the ones in the control group for fibroblasts and macrophages (p < 0.0001, p < 0.01, respectively). The difference between the two treatment groups was statistically significant for fibroblasts (p < 0.0001), but not significant for macrophages (p > 0.05). CONCLUSION: In spite of the fact that amniotic membrane transplantation is less effective than mitomycin C for depressing wound healing after trabeculectomy, it might be a safer alternative in selected cases when considering severe adverse effects of antimetabolites. PMID- 12566890 TI - Efficacy of latanoprost additive therapy on uncontrolled glaucoma. AB - PURPOSE: To assess the efficacy of latanoprost as additive therapy in patients with open-angle glaucoma and an intraocular pressure (IOP) deemed to be too high on maximum tolerated medical therapy. STUDY DESIGN: Prospective case series. METHODS: Consecutive patients with open-angle glaucoma, presenting to the Gulhane Military Medical Hospital Ophthalmology Clinic from May 1999 to September 2000 were enrolled. The effect of latanoprost on IOP was followed during a period of 12 months. The criterion for success was defined as having an IOP reduction of at least 20% from baseline or a final IOP of less than 22 mm Hg. Several clinical pretreatment variables (age, gender, ocular laterality, type of glaucoma, number of antiglaucomatous medications at study entry, pretreatment IOP) were analyzed for a significant effect on the efficacy of latanoprost additive therapy. MAIN OUTCOME MEASURE: IOP. RESULTS: Sixty-five eyes of 35 patients were included. The mean baseline IOP +/- SD was 23.3 +/- 2.0 mm Hg. Two patients (5.71%) developed ocular allergy in the first month requiring cessation of latanoprost. In the remaining 61 eyes of 33 patients, IOP was significantly reduced compared with baseline measurements with a mean IOP reduction of 6.1 +/- 1.8 (26.1%), 6.0 +/- 2.2 (25.3%) and 5.5 +/- 2.4 (23.2%) mm Hg at the 1-, 3- and 6-month follow-up controls, respectively (p < 0.001). Successful outcome was obtained in 50 (76.9%), 46 (70.7%) and 38 (58.4%) of 65 eyes at the 1-, 3- and 6-month visits, respectively. During the period from 6 to 12 months, 28 eyes underwent either a combined procedure (cataract extraction + intraocular lens implantation + trabeculectomy; 8 eyes) or only trabeculectomy (20 eyes) because of uncontrolled IOP; 4 eyes underwent the combined procedure because of visually significant cataract, and 8 eyes were lost to follow-up. Sixteen out of 21 eyes followed for more than 12 months with the same medications continued to have a successful outcome, and the mean IOP of 18.8 +/- 3.7 mm Hg was significantly different from baseline (p < 0.001). None of the pretreatment variables was a significant prognostic factor for failure of latanoprost additive therapy. CONCLUSION: This study supports the use of latanoprost additive therapy in patients with elevated IOP already receiving maximum-tolerated medical therapy. PMID- 12566891 TI - Timolol 0.1% gel (Nyogel 0.1% once daily versus conventional timolol 0.5% solution twice daily: a comparison of efficacy and safety. AB - In a prospective, randomised, double-masked, parallel-group, multi-centre study, 210 patients with primary open angle glaucoma or ocular hypertension were enrolled of whom 167 (timolol 0.1% gel 82, timolol 0.5% 85) completed the study as per protocol. The change in intraocular pressure between baseline and week 12 in the worse eye ('at trough') was 6.3 (SD 3.3) mm Hg on timolol 0.1% gel and 7.0 (2.9) mm Hg on timolol 0.5%; this difference was not statistically significant (p = 0.19). The difference between the two study groups in the change of intraocular pressure from baseline was 0.62 mm Hg; the 90% CI of -0.09 to +1.33 mm Hg was within the pre-specified limits of -1.5 to +1.5 mm Hg demonstrating equivalence between timolol 0.1% gel and timolol 0.5%. The plasma levels of timolol (ng/ml) at 12 weeks in the timolol 0.1% gel group were significantly less than that with timolol 0.5% both before instillation (mean 0.057, SD 0.131 and mean 0.470, SD 0.519 respectively, p = 0.025) and after instillation (mean 0.552, SD 0.992 and mean 2.473, SD 1.780 respectively, p = 0.008). Both treatments were well tolerated with no statistically significant difference between the groups in the occurrence of ocular or systemic adverse events. PMID- 12566892 TI - Effects of sildenafil citrate (Viagra) on choroidal congestion. AB - This study evaluates the effect of sildenafil on choroidal vascular congestion and its correlation with visual effects. Thirteen healthy subjects were randomized to a sildenafil group (n = 7, 3 M, 4 F), who received 200 mg of sildenafil, and a control (n = 6, 5 F, 1 M) group, who received no drug. Measurements of choroidal thickness with ultrasonography, color vision with Desaturated Panel D-15 Test, and contrast sensitivity with CSV-1000e charts (Vector Vision) were performed at baseline and at 90 and 180 min. Mean choroidal thickness and contrast sensitivity did not change significantly relative to baseline in either group. However, the variance in differences between repeat and baseline measurements of choroidal thickness was significantly higher at 90 min (p = 0.003) in the sildenafil subjects. Color discrimination error scores increased after sildenafil but did not correlate with changes in choroidal thickness. An oral dose of 200 mg of sildenafil caused small inconsistent changes in choroidal thickness, which did not correlate with visual effects. PMID- 12566893 TI - Endothelin-A receptor antagonist BQ-485 protects against intraocular pressure spike induced by laser trabeculoplasty in the rabbit. AB - PURPOSE: We have previously shown in rabbits that the intraocular pressure (IOP) spike caused by argon laser trabeculoplasty (ALT) is associated with an acute endothelin-1 (ET-1) release from the uveal tissue into the aqueous humour. In this study we investigated whether pretreatment with an ET(A) receptor antagonist (BQ-485) protects against the pressure spike induced by ALT, in the rabbit model. METHODS: Under general anaesthesia, 10 microl of 10(-5) M BQ-485 was injected into the anterior chamber of the right eye, and 10 microl of balanced salt solution (BSS) into the contralateral anterior chamber, for 12 pigmented rabbits. Five minutes later ALT (1,000 mW, 0.1 s, 100 spots over 360 degrees focused on the iris pillars) was performed on both eyes of each animal. IOP was measured before the injections (baseline value), and also 30 min afterwards using a Tono Pen XL tonometer. Immediately after the second IOP measurement aqueous humour was aspirated for measurement of ET-1 concentration. RESULTS: The baseline IOP (mean +/- SD) was 8.08 +/- 1.73 mm Hg and 7.92 +/- 1.78 mm Hg for the right and left eyes, respectively (Duncan test, p > 0.05). At 25 min after ALT the IOP of the BQ 485-pretreated right eyes remained unchanged (7.83 +/- 2.44 mm Hg, p > 0.05) but the IOP of the BSS-pretreated left eyes at 30 min increased significantly to 10.67 +/- 4.70 mm Hg (p < 0.05 for comparisons both with the corresponding baseline value for the same eye, and with the IOP of the contralateral eye at 30 min). There was no difference in aqueous humour ET-1 concentration between the corresponding right and left eyes (paired t test, p > 0.05). CONCLUSION: Intracameral BQ-485 pretreatment protected against the ALT-induced acute IOP elevation, but did not influence the laser-induced ET-1 release. This suggests that ET(A )receptor antagonists may potentially have a therapeutic role in the prevention of laser-induced IOP spikes. PMID- 12566894 TI - Comparison of 1-hour and 3-hours paclitaxel infusion pharmacokinetics: results from a randomized trial. AB - PURPOSE: Aim of this study was to characterize the difference in pharmacokinetics (PK) of paclitaxel (PAC) after 1-h and 3-h infusion in humans and to define a pharmacodynamic relationship between PAC PK and myelotoxicity. PATIENTS AND METHODS: PAC PK were studied during the first PAC application in the first treatment cycle (1 treatment cycle = 6 PAC applications) in 25 patients. This patient group represents a subgroup of a large clinical study with neurotoxicity as primary endpoint. These 25 patients were those patients who were willing to give additional blood samples. The group size was sufficient for a full description of the PK of PAC. PAC was administered at 100 mg/m(2) weekly by 1-h (n = 12) or 3-h (n = 13) infusion to patients with advanced cancer (lung, breast, ovarian, cervix, and head and neck). Total PAC was quantified by high-performance liquid chromatography (HPLC). Pharmacokinetic parameters were calculated by noncompartmental and model-dependent methods. The leukocyte and neutrophil decrease during a 6-week treatment period was calculated by the percentage in decrease of white blood cell count (WBC) and absolute neutrophil count (ANC) as well as the area over the curve (AOC) of WBC and ANC. RESULTS: The area under the curve (AUC), the plasma clearance (Clp), the volume of distribution at steady state (V(ss)), the mean residence time (MRT) and the distribution half-life (t(1/2)) of PAC(tot) were not different in the two application modes. The elimination half-life (t1/2) and maximum plasma concentration C(max) were significantly different. No significant differences in the percentage of reduction of WBC and ANC were seen. Calculation of AOC of WBC showed a borderline significant difference (p = 0.0547) in case of WBC and no significant difference in case of ANC between the two PAC schedules. A considerable variance of AOC was observed. CONCLUSION: The pharmacokinetic study of total PAC of the two schedules investigated showed significant differences in the elimination half-life, which is longer in case of the 1-h infusion of PAC and in the maximum plasma concentration, which is higher in case of the 1-h infusion. The two schedules showed a similar myelotoxicity with a trend of less toxicity in the 1-h procedure. PMID- 12566895 TI - The Ras-Raf-MEK-ERK pathway in the treatment of cancer. AB - The mitogen activated protein kinases (MAPKs) are conserved proteins that regulate cell growth, division and death. Although activated in the cytosol, the MAPKs translocate to the nucleus upon activation and phosphorylate a large number of nuclear proteins. Investigating how Ras transmits extracellular growth signals, the MAPK pathway has emerged as the crucial route between membrane-bound Ras and the nucleus. The MAPK pathway represents a cascade of phosphorylation events including three pivotal kinases, namely Raf, MEK (MAP kinase kinase), and ERK (MAP kinase). These kinases present new opportunities for the development of novel anti-cancer drugs designed to be target-specific and probably less toxic than conventional chemotherapeutic agents. A number of drugs inhibiting Ras, Raf or MEK are currently under clinical investigation. This review addresses the rationale for targeting the MAP kinase pathway and the current status of various pharmacological approaches. PMID- 12566896 TI - Antiangiogenesis: current clinical data and future perspectives. AB - Neovascularization is a prerequisite for progressive growth of solid tumors and their metastases. This process is tightly regulated by a large number of proangiogenic and antiangiogenic factors such as VEGF, bFGF and matrix metalloproteinases. The inhibition of angiogenesis is an innovative therapeutic approach and could represent a powerful adjunct to traditional therapy of malignant tumors. Preclinical trials have been very successful but in clinical studies meaningful response rates could only be shown in some cases. This might indicate the existence of different angiogenic phenotypes in humans. It seems that at present only a part of the interactions between the angiogenic cytokines are known. In addition, new receptor/ligand systems which regulate the neovascularization are being described. This article presents an overview of the most important angiogenically active substances, preclinical and clinical data, surrogate markers as well as future perspectives. PMID- 12566897 TI - DNA-based tumor vaccines. AB - Tumor vaccination strategies have been increased over the past years. This increase began with the identification of tumor antigens recognized by the immune system. Better understanding of the immune system and increasing knowledge about the antigen presentation process and the role of dendritic cells have opened new therapeutic possibilities. DNA vaccines, already successfully used against viral antigens and covering a broad repertoire of epitopes, might also be of advantage in tumor immunotherapy. Design and selection of vectors are of considerable importance for the vaccination. There are three major types of DNA-based recombinant cancer vaccines: DNA from tumor antigens can be used 1) to modify dendritic cells, 2) as 'naked' DNA-vaccine or 3) to construct recombinant viral vaccines. The current status in this field is shown and different vaccination strategies are discussed. PMID- 12566898 TI - The proteasome, a new target for cancer therapy. AB - The proteasome is a multicatalytic protease and the principal non-lysosomal proteolytic system in all eukaryotic cells. It plays a central role in virtually all regulatory pathways as for instance cell-cycle regulation, differentiation, and apoptosis. The proteasome degrades regulatory proteins and their inhibitors and, thus is an interesting target for therapeutic drugs. Inhibitors of the proteasome are small molecules that function by stabilizing various proteins, including cell-cycle regulators, tumor suppressors and growth factors. Because proteasome inhibition blocks cellular proliferation and induces apoptosis, these agents have been tested as anticancer drugs in tumor models and have shown impressive potential. In addition, treatment with proteasome inhibitors can sensitize cells to other cancer treatments like radio- or chemotherapy. This review introduces the ubiquitin-proteasome pathway and outlines the recent progress in the development of proteasome inhibition as a treatment option for clinical cancer therapy. PMID- 12566899 TI - Oral trofosfamide: an active and well-tolerated maintenance therapy for adult patients with advanced bone and soft tissue sarcomas. Results of a retrospective analysis. AB - OBJECTIVES: Aim of this study was to evaluate the feasibility and toxicity of oral trofosfamide given as maintenance therapy to adult patients with bone and soft tissue sarcomas following first or later line induction chemotherapy, and to determine the clinical efficacy in terms of impact on progression-free and overall survival. PATIENTS AND METHODS: 49 patients with locally advanced or metastatic high-grade soft tissue and bone sarcomas were identified retrospectively according to the inclusion criteria of the analysis. They were treated with oral trofosfamide at a dose of 100-150 mg per day continuously. All patients were pretreated with one or more lines of chemotherapy resulting in partial remission or stable disease. Patients were treated until progression of disease or unacceptable toxicity occurred. Progression- free and overall survival were measured from the beginning of maintenance therapy. RESULTS: Median follow up for all patients was 33 months (range 10-98). Toxicity was mild and predominantly hematologic. Only 1 patient had to stop treatment due to renal toxicity. The median progression-free survival was 7 months with 27% of patients continuing maintenance treatment at 1 year. Median overall survival is 14 months. Patients with metastatic disease showed a median survival of 23 months from diagnosis of metastases. 3 patients with stable disease following induction chemotherapy reached partial remission while under trofosfamide maintenance. CONCLUSION: Oral maintenance therapy with trofosfamide is well-tolerated and seems to prolong progression-free and overall survival compared to the course of advanced soft tissue and bone sarcomas without maintenance chemotherapy. PMID- 12566900 TI - Influence of age on the prognosis of patients with renal cell carcinoma (RCC). AB - OBJECTIVE: Aim of this study was to analyze the prognostic value of age in patients with renal cell carcinoma (RCC). PATIENTS AND METHODS: A group of 15 patients (age < or = 40 years, group I) and a group of 103 patients (age > or = 50 years, group II) with sporadic RCC who underwent radical nephrectomy between 1985 and 1997 were compared. The two groups were analyzed with respect to histologic cell type, tumor grade, stage and outcome. RESULTS: In group I low stage tumors (stage I and II) were diagnosed in 93% of patients and in group II in 65% of the patients (p = 0.017). High-grade tumors (stage III and IV) were diagnosed in 7% and 35% of patients in group I and group II, respectively (p < 0.01). In group I only one patient (7%) with stage II disease died of cancer. In group II the distribution of cancer-specific mortality was as follows: 6 patients (100%) with stage IV, 13 patients (42%) with stage III, and 12 patients (17%) with stage I and II died of disease. The 5-year-survival in group I was 93% and in group II was 77% (p = 0.05). CONCLUSION: According to our data we conclude that RCC is diagnosed at a lower stage in young patients than in the older patient group. RCC may exhibit a more favorable prognosis in young patients, possible due to the lower stage at time of diagnosis. PMID- 12566901 TI - Surgery of low-grade gliomas near speech-eloquent regions: brainmapping versus preoperative functional imaging. AB - The identification of eloquent areas is of utmost importance in the surgery of tumors located near speech-eloquent brain areas, since the classical concept of a constant localization was proven to be untrue and the spatial localization of these areas may show large interindividual differences. Some neurosurgical centers apply intraoperative electrophysiological methods that, however, necessitate the performance of surgery in the awake patient. This might be a severe burden both for the patient and the operating team in a procedure that lasts several hours; in addition, electrical stimulation may generate epileptic seizures. Alternatively, methods of functional brain imaging (e.g., PET, fMRI, MEG) may be applied, which allow individual localization of speech-eloquent areas. Matching of these image data with a conventional 3D-CT or MRI now allows the exact transfer of this information into the surgical field by neuronavigation. Whereas standards concerning electrophysiological stimulation techniques that could prevent a permanent postoperative worsening of language are available, until now it remains unclear whether the resection of regions shown to be active in functional brain imaging will cause a permanent postoperative deficit. PMID- 12566902 TI - Postradiation high-grade myofibroblastic sarcoma of the prostate -- a rare entity of prostatic tumors -- responding to liposomal Doxorubicin. AB - BACKGROUND: Prostatic postradiation sarcoma (PRS) is a very rare malignant disease with a dismal prognosis. Aggressive surgical resection is the most widely used therapy for PRS. Chemotherapy so far has been unsatisfactory, with doxorubicin being the only drug with established activity. Liposomal doxorubicin has been shown to have comparable activity and a more favorable toxicity profile compared with unpegylated doxorubicin. CASE REPORT: A 78-year-old man presented with high-grade myofibroblastic sarcoma 13 years after curatively intended pion irradiation of the prostate with 30 Gy. He was treated with liposomal doxorubicin 40 mg/m(2) every 4-6 weeks up to now with a total dose of 800 mg. Partial remission has been achieved after 17 months of treatment with liposomal doxorubicin. Toxicity is minimal with palmoplantar erythrodysesthesia WHO grade 1, resolving completely after extending treatment intervals to 6 weeks. RESULT: Liposomal doxorubicin led to a sustained tumor stabilization over 17 months in a patient with prostatic postradiation high-grade sarcoma. Postradiation sarcoma (PRS) usually has a dismal prognosis and only very limited therapeutic possibilities. As in sarcomas of different origin, anthracyclines including liposomal doxorubicin seem to have potential antitumor activity in postradiation sarcoma of the prostate. PMID- 12566903 TI - Classification of malignant lymphoma. AB - Over the past 30 years different lymphoma classifications were used world-wide. In Europe, the Kiel classification of K. Lennert dominated the scene, it was updated 1988 and 1992. In North America the Working Formulation, primarily designed to be an instrument for translation of one classification into the other, was predominantly used. Both classifications, however, showed little correspondence with each other, which made it difficult to compare the results of randomized clinical trials on both sides of the Atlantic Ocean. In 1994 the Revised European American Lymphoma (R.E.A.L.) classification was proposed by the International Lymphomas Study Group (ILSG). In contrast to the existing classifications it abandoned the grading of malignancies. The ILSG focused on diagnostic reproducibility and on the definition of distinct clinical pathological lymphoma entities. Similar to the Kiel classification the new classification scheme was based on cell lineage (T- and B-cell origin) and cell differentiation (precursor and mature lymphomas) and included a number of distinct extranodal lymphomas. The new WHO classification is basically identical with the R.E.A.L. scheme and exhibits only minor changes. The WHO classification includes all hematopoietic and lymphoid neoplasms. It represents the first generally accepted classification, providing hematologists and oncologists with a solid diagnostic basis for therapeutic decisions. PMID- 12566904 TI - Sexuality and sexual dysfunction in gynecological psychooncology. PMID- 12566905 TI - Vinorelbine and carboplatin in inoperable non-small cell lung cancer: a monoinstitutional phase II study. AB - Chemotherapy regimens for patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) provide typically palliation or limited symptom-free survival. We investigated the efficacy, in terms of response rate and time to progression, of a combination chemotherapy containing carboplatin and vinorelbine as a first-line treatment for inoperable NSCLC. Fifty-two consecutive patients with advanced NSCLC were treated with carboplatin AUC 6 according to Calvert's formula on day 1, combined with vinorelbine, 25 mg/m(2) on days 1 and 8. Therapy was given every 3 weeks. The median age was 66 years (range, 40-80); ECOG performance status was 0 in 20, 1 in 25 and 2 in 7 patients. According to an intent-to-treat analysis, response rate (partial and complete responses) in 52 assessable patients was 18/52 (34.6%; 95% confidence interval, 22-47%). In addition, 16 patients (30.8%) had stable disease and 18/52 (34.6%; 95% CI, 22 47%) progressed while on treatment. Median time to progression and overall survival were 7, 5 and 12.3 months, respectively. Grade 3/4 granulocytopenia was observed in 18/8 patients (34.6/15.4%). Grade 3/4 nadirs generally lasted no more than 7 days, and no neutropenic fever was reported. The treatment was generally very well tolerated: grade 1 or 2 nausea and vomiting was observed in 12 and 4 patients, respectively, and grade 2 neuropathy in 5% of cases. Statistical analysis did not highlight any significant differences in clinical benefit (partial and complete responses and stable disease), time to progression, or grade 3-4 hematologic and non-hematological toxicity according to age (65 years). Carboplatin AUC 6 and vinorelbine was found to be an efficacious regimen as a first-line treatment for inoperable lung cancer patients and was also subjectively very well tolerated in aged patients. The regimen warrants further investigation in the emerging subgroup of aged patients in order to draw firm conclusions. PMID- 12566906 TI - Prevention of anemia in patients with solid tumors receiving platinum-based chemotherapy by recombinant human Erythropoietin (rHuEpo): a prospective, open label, randomized trial by the Hellenic Cooperative Oncology Group. AB - OBJECTIVES: Platinum compounds are commonly associated with significant anemia. Erythropoietin administration has been found effective in correcting anemia in patients with solid tumors receiving chemotherapy. We conducted a randomized, open label study to assess the efficacy of erythropoietin in preventing transfusions and significant anemia (hemoglobin <10 g/dl) in patients with solid tumors receiving platinum-based chemotherapy. METHODS: One hundred forty-four patients with hemoglobin <13 g/dl were included in this study (72 in each arm). Patients in the treatment arm received 10,000 U of recombinant human erythropoietin (rHuEPO) thrice weekly s.c. during platinum-based chemotherapy, while patients in the control arm received no treatment. RESULTS: All patients were evaluable for efficacy. Transfusions were reduced by the administration of rHuEPO (15.3 vs. 33.3%, p = 0.019), and fewer patients developed significant anemia (16.6 vs. 45.8%, p < 0.0001). Subgroup analysis showed that patients with observed to predicted (O/P) serum erythropoietin levels 0.9 or non-responders. CONCLUSIONS: rHuEPO at a dose of 10,000 U thrice weekly prevents transfusions and development of significant anemia in patients with solid tumors receiving platinum-based chemotherapy. PMID- 12566907 TI - Pharmacokinetic study of two infusion schedules of irinotecan combined with cisplatin in patients with advanced gastric cancer. AB - OBJECTIVE: Irinotecan (CPT-11) in combination with cisplatin (CDDP) has shown promising antitumor activity for advanced gastric cancer, but the optimal administration schedule of CPT-11 is still controversial. To clarify the pharmacokinetic effects of different CPT-11 administration schedules, we compared two different regimens (continuous infusion of CPT-11 for 24 h and CPT-11 infusion for 90 min) combined with CDDP in patients with advanced gastric cancer. PATIENTS AND METHODS: Five patients were treated with CPT-11 at a dose of 60 mg/m(2) delivered by continuous infusion for 24 h on day 1 and by a 90-min infusion on day 15, together with CDDP daily administered at a dose of 10 mg/m(2) on days 1-3 and days 15-17 for 4 weeks. The pharmacokinetics of CPT-11 and its metabolites, SN-38 and SN-38 glucuronide (SN-38G), were investigated, as well as the toxicity of therapy. RESULTS: Grade 3 leukopenia was observed in 1 patient after 24-hour infusion and in 1 patient after 90-min infusion of CPT-11. In addition, grade 3 thrombocytopenia was observed in 1 patient after the 90-min infusion. Other adverse reactions were mild, and the planned dose was delivered to all patients. The area under the plasma concentration-time curve of SN-38, the active metabolite from CPT-11, was increased by 24-hour infusion when compared with the 90-min infusion, and there was no increase in toxicity. CONCLUSION: Protracted infusional CPT-11 combined with CDDP is a practical regimen, and may be appropriate for a future phase II trial. PMID- 12566908 TI - A case-control study of endometrial cancer after antipsychotics exposure in premenopausal women. AB - OBJECTIVE: Most endometrial cancers are related to hormonal imbalance, and antipsychotics are a common cause of hyperprolactinemia. We investigated the possible relation between the use of antipsychotics and the risk of endometrial cancer. METHODS: A case-control study was conducted on premenopausal women at the Chiba University Hospital between 1989 and 2000. The cases were 41 patients with histologically confirmed endometrial cancer. For each case, there were 3 controls matched by age and date of visit. Subsequently 123 women without endometrial cancer were enrolled as controls. Detailed information including obesity, parity, hypertension, diabetes mellitus, use of exogenous estrogen and use of antipsychotics was analyzed. Odds ratios, two-sided p values and 95% confidence intervals were estimated by a conditional logistic regression analysis. Radioimmunoassays of serum luteinizing hormone, follicle-stimulating hormone, prolactin, estradiol and progesterone were performed in 4 of 5 patients with endometrial cancer who were using antipsychotics. RESULTS: Use of antipsychotics, diabetes mellitus and obesity were identified as independent variables with risk estimates of 5.4, 9.3 and 4.9, respectively. Serum progesterone, estradiol and prolactin levels ranged from 0.4 to 0.7 ng/ml, 32 to 110 pg/ml and 34 to 258 ng/ml, respectively. CONCLUSIONS: Use of antipsychotics is a risk factor for endometrial cancer, and hyperprolactinemia associated with antipsychotics may impart the risk of endometrial cancer in premenopausal women. PMID- 12566909 TI - Phase II study of concurrent administration of doxorubicin and docetaxel as first line chemotherapy for metastatic breast cancer. AB - OBJECTIVE: To evaluate the efficacy and toxicity of concurrent administration of doxorubicin and docetaxel, without prophylactic use of granulocyte colony stimulating factor, as first-line chemotherapy in patients with metastatic breast cancer (MBC). METHODS: This multi-institutional study enrolled 40 women; 37 were assessable for efficacy and all 40 patients were evaluated for toxicity. Treatment consisted of 50 mg/m(2) doxorubicin and 60 mg/m(2) docetaxel on day 1 every 3-4 weeks. RESULTS: Patients received a total of 251 cycles of chemotherapy (median, 5 cycles; range, 1-13 cycles). Of the 37 patients assessable for efficacy, 2 had a complete response and 24 had partial responses, which accounted for a 70% objective response rate (95% confidence interval, 53-84%). The median time to treatment failure was 30.1 weeks (range, 3.3-80.7 weeks). Grade 4 neutropenia was observed in 88% of patients and was the most frequent haematological toxicity. Febrile neutropenia was seen in 40% of patients, but no severe infections were observed. Non-haematological toxicity was generally tolerable. There were 2 grade 4 adverse events, which included 1 bleeding duodenal ulcer and 1 hypersensitivity reaction, but grade 3 episodes were infrequent. None of the patients developed congestive heart failure or asymptomatic decrease of left ventricular ejection fraction to less than 50%. Fluid retention syndrome 3 mm (1.3) and Apo(E-4) (2.1) were significant predictors of dementia. The combination of an Apo(E-4) genotype, 3MSE score <90, > or =5 ventricular grade, > or =3 white matter grade at the time of the MRI were associated with a 17-fold increased risk (95% CI: 8.6-34.9) of dementia as compared to individuals with none of the above attributes. CONCLUSIONS: Measures of cognition, Apo(E-4) and MRI of the brain are strong predictors of both dementia and of AD. PMID- 12566950 TI - Changes in Mini-Mental State Exam in community-dwelling older persons over 6 years: relationship to health and neuropsychological measures. AB - Although cognitive screening test scores change with advanced age, the significance of these changes (particularly decline in score) needs to be defined in terms of general health and neuropsychological functioning. Such analysis was undertaken in a subgroup of 287 healthy older men (mean age at baseline = 70.7 years) from the Western Collaborative Group Study, an ongoing cardiovascular and aging research project. Time from baseline to follow-up study averaged 6.0 years (SD = 0.5 years). Mini-Mental State Examination (MMSE) scores indicated that 15% of participants declined by three or more points (a 1+ standard deviation change among all change scores), 5% of participants improved by three or more points, and 80% of the sample remained within two points of their initial score. In health terms, decliners were significantly older, less active at follow-up, rated their health more poorly, and reported more depressive symptoms than non decliners. Decliners also performed more poorly on several neuropsychological tests administered at follow-up. Results suggest that a decline of three or more points on the MMSE in community-dwelling, older persons without acute illness may signify important changes in health and cognition. PMID- 12566951 TI - A brief descriptive analysis of stroke features in a population of patients from a large urban hospital in Richmond, Virginia, a city within the 'stroke belt'. AB - PURPOSE: To provide a descriptive analysis of the stroke features in a population of patients from a large urban hospital within the 'stroke belt', with the intention of stimulating interest in creating similar comprehensive databases across the country; the differential population features that may contribute to the increased incidence of ischemic and hemorrhagic stroke within the 'stroke belt' can thus be better studied and understood. BACKGROUND: Strokes account for approximately 5-10% of all deaths in the western world. Within a geographically nondescript region in the Southern United States known as the 'stroke belt', however, the incidence of stroke is 1.5-2 times greater than in other regions of the country. Despite the relatively high incidence of stroke within the 'stroke belt', very little information is available regarding the differential population features that distinguish stroke patients within this region from those in other parts of the country. DESIGN/METHODS: During the period September 1, 1995 to August 31, 1998, data for 506 patients meeting criteria for stroke as outlined in the National Survey of Stroke were entered into the 'Adult Brain Attack Database' at the Medical College of Virginia Hospitals of Virginia Commonwealth University. The data were analyzed using descriptive statistics. RESULTS: Of the 506 patients entered into the database 281 experienced their first stroke during the data collection period. Of these individuals, 60.5% (n = 170) were female (mean age 60.6). The mean age in men presenting with new stroke (n = 111) was 62.5. African Americans represented the predominant ethnic group making up 63.3% (n = 178) of all first stroke patients. The three most common risk factors among deceased ischemic stroke patients (3.2%; n = 9) included hypertension (58.3%), alcohol abuse (41.6%) and cigarette smoking (33.3%). Among ischemic stroke survivors, the three most common risk factors included hypertension (53.6%), prior neurovascular disease (36.2%) and cigarette smoking (28.0%). Risk factors among hemorrhagic stroke patients included hypertension (42.0%), cigarette smoking (26.3%) and alcohol abuse (21.1%). The most common complication associated with stroke across all subtypes was hypertension with an overall prevalence of 9.3%. The use of antiplatelet and antihypertensive therapy at discharge among stroke patients in whom such data was recorded was 58.8% (179/289) and 48.9% (132/270), respectively. DISCUSSION: Future studies comparing data from the Richmond, Va. stroke database with data from other regions of the country outside the 'stroke belt' may allow for differentiation of the population features that contribute to the increased prevalence of stroke within the 'stroke belt'. PMID- 12566952 TI - Risk factors for atherosclerosis of cervicocerebral arteries: intracranial versus extracranial. AB - OBJECTIVES: To search for factors determining the site of atherosclerosis of the cervicocerebral arteries in patients from Bangkok (Thailand) with ischemic stroke in the carotid territory. MATERIAL AND METHODS: Patients with the clinical syndrome of stroke or transient ischemic attack in the carotid territory or with ocular stroke were retrospectively studied. They were divided into an intracranial and an extracranial carotid stenosis group based on their vascular imaging. The stroke risk factors were then compared. RESULTS: There were 49 cases with extracranial carotid stenosis and 51 with intracranial stenosis. Among the patients with extracranial stenosis, 98% had associated intracranial disease, whereas none of those with intracranial stenosis had more than 50% of extracranial carotid stenosis. The presence of diabetes mellitus and a history of ischemic heart disease were found to be significantly more prevalent among patients with extracranial internal carotid artery stenosis. CONCLUSION: Diabetes mellitus as well as a history of ischemic heart disease were found to be more significantly prevalent in patients with combined extracranial internal carotid artery and intracranial stenosis. Extracranial carotid stenosis might represent a more severe atherosclerotic process of the cervicocerebral circulation when compared with intracranial stenosis. PMID- 12566953 TI - Incidence of Parkinson's disease in estonia. AB - The incidence of Parkinson's disease (PD) was studied over a 9-year period (1990 1998) in Tartu county, Estonia, with a mean population of 156,417. Based on 264 patients, the average crude incidence rate of PD was 18.8 per 100,000 person years. The age-adjusted incidence was 16.8/100,000; 18.3 for the urban group and 14.0 for the rural group; 16.6 for men and 17.1 for women. The age-specific incidence was 1.0/100,000 in the age range 30-39 years, reached a maximum of 117.2/100,000 in the age range 70-79 years and declined in the elderly. The incidence of PD was comparable with that found in developed countries, except for a slightly higher (not significant) incidence rate in the urban compared with the rural population in Estonia. PMID- 12566954 TI - Brain cancer in a residential area bordering on an oil refinery. AB - Both in response to community concerns about brain cancer related to an oil refinery and in order to more fully understand the etiology of primary site brain cancer (glioma), a highly focused cancer cluster investigation was conducted. The components included: (1) a literature review of occupational exposures in the petroleum refining and petrochemical industries, (2) comparisons between observed and expected cases, (3) comparisons between mean age at diagnosis and median survival time and (4) interviews concerning exposures of cases. Evidence from the literature review revealed little, if any, effect of petroleum refinery or petrochemical exposure on the risk for brain cancer. There was no statistically significant increase in the number of brain cancer cases in the community (observed = 12, expected = 9.46, standardized mortality ratio = 1.27). There was no statistically significant decrease in mean age at diagnosis or median survival time among those most exposed. Reports of exposure from the case interviews were highest for eating processed meats (98.5%), dental X-rays (96.6%), dog ownership (91.2%) and swimming (80.3%). There were no major occupational exposures identified. It seems unlikely that petrochemicals are involved in any significant way in the etiology of most brain cancers (gliomas). A follow-up case-control study should focus primarily on those risk factors mentioned frequently by the cases. PMID- 12566955 TI - Projection of creutzfeldt-jakob disease frequency based on cadaveric dura transplantation in Japan. AB - BACKGROUND: Projections of the size of the Creutzfeldt-Jakob disease (CJD) epidemic are of great importance for future health care and understanding of the natural history of this disease. OBJECTIVE: We have estimated the incubation period and have projected the number of patients with iatrogenic CJD in Japan, using 75 patients who were transplanted with cadaveric dura mater grafts between 1979 and 1991 and developed CJD by the end of 2000. METHODS: Several plausible scenarios, combinations of incubation period distribution forms, competing risks and exposure distributions were evaluated for the projections using data from the Japanese Ministry of Health, Labor and Welfare. RESULTS: The range of median incubation periods from the time of dura mater transplantation to the onset of the disease was 10.8-34.3 years, and the range of the number of new CJD patients between 2001 and 2020 was 21-84. CONCLUSIONS: Since the potential incubation period was estimated to be considerably long, as few as 21 and as many as 84 new patients may develop CJD by 2020. PMID- 12566956 TI - The use of standardized incidence and prevalence rates in epidemiological studies on multiple sclerosis. A meta-analysis study. AB - OBJECTIVE: To demonstrate whether or not the age and sex adjustment of incidence and prevalence rates in multiple sclerosis (MS) could allow more reliable comparison between epidemiological studies performed in different areas of the world and to establish if the latitude gradient theory could be confirmed after the standardization for age and sex distribution. METHODS: A meta-analysis of population-based incidence and prevalence studies on MS from 1980 through 1998 using the terms 'multiple sclerosis', 'prevalence' and 'incidence' in the bibliographic databases MEDLINE and EMBASE was performed. We included studies that reported the diagnostic criteria, number of cases and the population studied, the date of the study, the latitude, and the age- and sex-specific crude incidence and prevalence rates. According to the inclusion criteria, 69 of 127 papers on prevalence and 22 of 70 papers on incidence were considered for age adjustment and 27 prevalence and 8 incidence studies for sex adjustment. The mean incidence and prevalencerates and the 95% confidence intervals age- and sex adjusted to the World and the European standard populations were calculated. RESULTS: The Spearman rank correlation and the multiple regression analyses indicated that age adjustment to standard populations could overcome the limitations in comparing the crude prevalence and incidence rates of different epidemiological studies on MS. When the mean crude and age- and sex-adjusted prevalence and age-adjusted incidence rates were stratified by latitude (from south to north), the latitudinal gradient, which was highly significant for the crude rates, became less remarkable for the age- and sex-adjusted prevalence rates and not significant for the age-adjusted incidence rates. CONCLUSIONS: The crude incidence and prevalence rates in epidemiological studies on MS should be age- and sex-adjusted to a common standard population to permit a more reliable comparison among studies performed in different countries. Our findings support the opinion that the latitude does not play a key role in determining the onset of MS. Whenever possible, the crude incidence and prevalence rates should be adjusted to the ethnic origin and migration characteristics. PMID- 12566957 TI - Geographic and temporal distribution of mortality rates for multiple sclerosis in Canada, 1965-1994. AB - Statistics Canada data were used to calculate multiple sclerosis (MS) mortality rates per 100,000 population in the Canadian provinces from 1965 to 1994. For the period 1965-1994, the highest average annual MS mortality rates were in Quebec (4.4) and Ontario (3.9), while the Western Provinces had an intermediate rate (2.1) and the Atlantic Provinces had the lowest rate (1.2). Female mortality rates exceeded male rates in each of the four regions. Average annual MS mortality rates in Canada overall fluctuated during the past 30 years, with rates of 3.4 in 1965-1969, 4.2 in 1970-1974, 3.2 in 1975-1979, 2.3 in 1980-1984, 2.8 in 1985-1989 and 3.9 in 1990-1994. Female mortality rates exceeded male rates during each 5-year period. The highest mortality rates for both genders were in the 65 years plus age group. Rates in the under 45 years age group have remained stable, while rates in both the 45-64 and 65 years plus age groups have fluctuated. There is no apparent relationship between prevalence and mortality rates among the Canadian provinces. PMID- 12566958 TI - The frequency of multiple sclerosis in jewish and arab populations in greater jerusalem. AB - A comparison of the incidence rate (IR) and the prevalence rate (PR) of multiple sclerosis (MS) in subgroups of the same ethnic origin, but born and living in different geographical areas, may delineate the relationship between environmental and genetic risk factors for MS. Previous epidemiological studies of MS in Israel did not include the Arab population and used diagnostic criteria that did not include MRI findings. Therefore, we studied the age-adjusted IR and PR of MS in a more recent sample in different population groups, including Arabs, of Greater Jerusalem. We found that the PR of MS in Israeli Jews is higher than previously described. Furthermore, the PR was significantly lower among immigrant Jews from Asia/Africa (A/A) than among native-born Jews of Asian/African origin (I-A/A). Since these groups have similar genetic susceptibilities to MS, the higher PR in the latter is probably due to environmental factors. Our study does not support the effect of latitude on the risk of developing MS since no difference in the PR was found between immigrant Jews from Europe/America (E/A) and native-born Jews of European/American origin (I-E/A). Among Arabs, the PR was similar to that among A/A. Therefore, we hypothesized similarity in environmental etiologic factors for MS between the countries of origin of A/A immigrants and of Arabs communities in Greater Jerusalem. The IR of I-E/A was higher than that of I A/A and Arabs, although this difference did not reach statistical significance. PMID- 12566960 TI - Ambient air quality and occurrence of multiple sclerosis relapse. AB - Infectious viruses and bacteria can trigger multiple sclerosis (MS) exacerbations. Seasonally changing concentrations of ambient air pollutants are known to predispose to transmissible infections, to induce systemic immune responses and to enhance existing peripheral inflammation. Ambient air quality and monthly MS relapse occurrence in south-western Finland were compared by multivariate logistic regression. The odds ratio of the risk of a relapse onset was over fourfold (4.143, p < 0.001) when the concentration of inhalable particulate matter (PM(10)) was at the highest quartile. Inhalable airborne particulate matter concentrations were connected to relapse occurrence. Poor air quality may enhance the seasonal changes in MS relapse occurrence by an increased susceptibility to transmissible infections. PMID- 12566959 TI - Radon: a possible risk factor in multiple sclerosis. AB - Ecological studies in Norway, using a method for spatially moving bivariate correlation analysis, show that south of 65 degrees N, there are significant positive correlations (p < 0.01) for rates of multiple sclerosis (MS) versus contents of radon (Rn) in indoor air, and significant negative correlations for MS rates versus fallout of magnesium (Mg) and amounts of precipitation. Based on these data, we propose the hypothesis that the content of Rn in inhaled air is a risk factor in MS. The release of harmful Rn levels to the air may be influenced by (1) the levels of exchangeable Mg in soil, which may affect the soil content of the Rn precursor radium (Ra), and (2) the amounts of precipitation through its effects on soil moisture, which is one of the factors controlling Rn emanation from the soil. This hypothesis agrees with several of the known epidemiological characteristics of MS. PMID- 12566961 TI - Effects of free radicals on coronary artery. AB - Coronary arteries supply blood to the heart and hence the control of coronary tone is pivotal to human survival. Reactive oxygen species (ROS) in specified amounts play an important role in normal metabolic and signalling processes. However, excess ROS can cause severe cardiovascular damage. For example, NO is produced by endothelium as a signal for relaxation. However, in an inflammatory response, NO from endothelium or macrophages can combine with superoxide to produce more deleterious peroxynitrite. Excess ROS have been associated with loss of coronary artery pliability--loss of contraction in some instances and relaxation in others. Atherosclerosis may also be considered an inflammatory response that leads to artery blockage, coronary disease and ischaemia reperfusion. ROS produce various types of damage to ion channels and pumps and this damage is associated with vascular diseases such as atherosclerosis and hypertension. Endothelium and smooth muscle in the coronary artery are also affected differently by individual ROS. In fact, endothelium may act to protect the underlying smooth muscle against ROS. This review will give an overview of this field. PMID- 12566962 TI - Nitric oxide production and arachidonic acid metabolism in platelet membranes of coronary heart disease patients with and without diabetes. AB - AIM: To evaluate the levels of nitrite (NO(-)(2)) and nitrate (NO(-)(3)) ions and the incorporation of [(3)H]arachidonic acid (AA) into phospholipids of platelet membranes from coronary artery disease (CAD) patients with and without diabetes (NIDDM). SUBJECTS AND METHODS: Eighteen CAD patients (group A), 18 CAD patients with NIDDM (group B), and 20 healthy controls (group C) without dyslipidemia, peripheral vascular disease and hypertension were included in the study. The groups were matched for age, sex and body mass index. The diagnosis of CAD was confirmed by coronary angiography. The nitric oxide end products (NOx), NO(-)(2) plus NO(-)(3) ions in platelet membranes, were determined using a spectrophotometric method based on the Griess reaction. The turnover of phospholipids was evaluated by incorporation of [(3)H]AA into platelet membrane phospholipids. RESULTS: A significantly smaller amount of NOx ions was in the platelet membrane of groups A (40 +/- 8 micromol/l) and B (29 +/- 10 micromol/l) than C (57 +/- 6 micromol/l), p < 0.001. Conversely a significantly greater amount of [(3)H]AA was incorporated into platelet phospholipids of group B patients (5,123 +/- 1,637 dpm/mg) than groups A (3,159 +/- 1,253 dpm/mg; p < 0.002) and C (1,621 +/- 417 dpm/mg). An inverse correlation between [(3)H]AA incorporation and NOx levels was established: r = -0.76 (p < 0.05, n = 36) in CAD patients. CONCLUSIONS: Diabetes in CAD patients decreased the ability to produce platelet-derived NO and affects AA metabolism. This may result in higher platelet sensitivity to aggregating stimuli. PMID- 12566963 TI - Lymphoscintigraphy in the sentinel lymph node technique for breast tumor: value of early and late images for the learning curve. AB - As the performance of early (H+1 to H+4) and late (D1) lymphoscintigraphic images raises organizational problems in outpatient surgery for breast cancer, only early images are generally obtained. The present study evaluated whether two series of images are better than one and defined the advantages of both methodologies. One hundred and eighteen patients with infiltrating breast carcinoma (T(0), T(1) and T(2)) were included in the study: 87 in group A (early and late images) and 31 in group B (only early images). All patients received two peritumoral injections of (99m)Tc-sulfur colloid, 15-18 MBq (group A) and <15 MBq (group B). During the operation, the patent blue bye technique was associated with radioactivity detection. The two groups were comparable for histological type and tumor size and localization. Successful localization of sentinel nodes on early lymphoscintigraphic images was significantly greater for group B. The identification of a sentinel node focus on early lymphoscintigraphy increased by 10% during the study. Sentinel node detection by the isotopic method alone, or the two methods combined, was comparable for both groups. In radioactivity detection, the count rate for sentinel nodes versus background (contralateral breast) was similar for the two groups. During the learning phase, two series of images gave a definite advantage. Subsequently, lymphoscintigraphy performed at +2 h was sufficient (the results for the two groups became indistinguishable). PMID- 12566964 TI - Mutations of the BRCA1 gene in hereditary breast and ovarian cancer in the Czech Republic. AB - OBJECTIVE: Mutations in the BRCA1 gene confer a high risk for the development of breast or ovarian cancer. The aim of this study was to establish the frequency and spectrum of BRCA1 mutations in Czech breast or ovarian cancer families. SUBJECTS AND METHODS: We have screened 28 patients with early-onset cancer and 54 patients from risk families for germ-line mutations in BRCA1. All coding exons were analyzed by the protein truncation test (PTT) and the reverse transcription polymerase chain reaction (RT-PCR). Detected variants were characterized by direct sequencing of PCR products. RESULTS: Five distinct deleterious mutations have been identified in six families. All the mutations led to the premature termination of translation. One mutation was detected in a group of 11 (9.1%) patients with early-onset breast cancer. Two mutations were observed in 32 (6.3%) families with a history of breast cancer only. Three mutations were found in 22 (13.6%) families with both breast and ovarian cancer. The 5382insC mutation was the only abnormality detected twice. In addition, two variant transcripts, the loss of exon 5 and in-frame 3-bp deletion at the beginning of exon 8, were identified only at the cDNA level. Their biological significance remains unknown. CONCLUSION: PTT analysis enables examination of long PCR products. The technique is useful for rapid detection of mutations in hereditary breast cancer. PMID- 12566965 TI - Deaths in patients with pulmonary tuberculosis: an analysis of a chest diseases hospital in Istanbul, Turkey. AB - OBJECTIVE: To evaluate the medical records of patients with pulmonary tuberculosis in order to determine the inhospital mortality rate. MATERIALS AND METHODS: Medical records of 22,651 patients with pulmonary tuberculosis admitted to Gulhane Military Medical Academia, Camlica Chest Diseases Hospital, Istanbul, Turkey from 1977 to 1999 were examined. RESULTS: Of the 22,651 patients, 133 (0.58%) died: 117 male and 16 female, mean age 41.5 +/- 20.7 years. The mean length of hospital stay was 15.8 +/- 25.9 days and it increased yearly (p = 0.004). There was no significant difference in hospital mortality rate of tuberculosis within years. Forty-nine patients had chronic and/or far advanced tuberculosis, 36 of them had concomitant diseases. Thirteen patients had miliary and/or tuberculous meningitis, 3 multi drug-resistant tuberculosis, another 3 had pneumothorax, and 2 empyema. These patients had significantly lower prognostic nutritional index than those patients who recovered from pulmonary tuberculosis (p = 0.025). CONCLUSION: Inhospital mortality rate of tuberculosis did not change in the last 23 years. We suggest that pulmonary tuberculosis will continue to be a significant cause of death and therefore important health care problem for Turkey in the 21st century. PMID- 12566966 TI - Vibration and its effects on the body. AB - OBJECTIVES: To assess the effects of machine-induced vibration on workers and to determine effective precautions for vibration-induced trauma. SUBJECTS AND METHODS: The study group consisted of 114 workers who were randomly selected: 50 rock drill workers and 64 truck heavy vehicle operators. Fifty-four office workers were designed as controls. The study and control groups were age-matched. All subjects were interviewed to determine subjective symptoms using a 38-item questionnaire designed by the Medical Committee of Vibration Disease, Japanese Association of Industrial Health. RESULTS: The complaints of pain in the fingers, sensitivity to cold, numbness and pain of fingers at night, weakness of static position, wrist-elbow pain, difficulty in bending and stretching elbow, pain in shoulder when holding up arms, lower back pain, sleeping disturbance and hearing difficulty were significantly higher in rock drillers than heavy vehicle operators and office workers (p < 0.05-0.01). CONCLUSION: Permanent vibration exposures cause negative physical effects that may lead to occupational diseases. In order to be protected against whole-body and hand-arm vibrations, technical and medical measures must be taken into account. PMID- 12566967 TI - Patient satisfaction with three dental speciality services: a centre-based study. AB - OBJECTIVE: To assess the level of patient satisfaction with three dental speciality services: oral surgery, periodontics, and fixed prosthodontics. SUBJECTS AND METHODS: A sample of 1,242 patients, aged 16 years and over who received dental care at least twice at the main dental centre, Al-Amiri, Kuwait, were interviewed during May-August 1998 by independent researchers concerning their experience of previous visits. Patients reported their level of satisfaction on a 5-point Likert scale for the items in the questionnaire, which focused on four areas: access, dentist's communication, staff helpfulness, and structural aspects of the practice. RESULTS: On average 49.4% (25.9-89.6%) of participating patients rated the level of care received as excellent. Areas rated poorest included waiting time for an appointment and waiting time in the clinic to access the dentist. Participants were less satisfied with doctor's explanation of illness, dental treatment and confidentiality of medical records. The areas that received excellent ratings by patients included: (a) communication (listening and understanding complaints 40.0% and courteous treatment 57.8%), and (b) structural aspects (availability of infection control and safety measures 89.6% and packed sterilized instruments 78.9%). CONCLUSION: Findings indicate that the level of patient satisfaction was reasonably high. Areas identified as needing improvement included: dentist-patient communication skills, medical records, appointment systems, and shortening of the waiting time to access the dentist. PMID- 12566968 TI - Nasogastric tube syndrome: the unilateral variant. AB - Nasogastric tube syndrome is an uncommon but a potentially life-threatening complication of nasogastric intubation. It presents as an acute upper airway obstruction secondary to bilateral abductor vocal cord dysfunction resulting from postcricoid chondritis. However, a unilateral variant has also been reported and may herald the development of bilateral pathology. The purpose of this report is to highlight early warning signals of impending crisis. CLINICAL PRESENTATION: A case of unilateral vocal cord involvement secondary to postcricoid chondritis resulting from nasogastric intubation is presented. An additional feature, not reported previously, is the presence of two inflammatory nodular swellings over the cricoid cartilage. PMID- 12566969 TI - Primary ankle fusion using blair technique for severely comminuted fracture of the talus. AB - OBJECTIVE: We report a case of a severely comminuted fracture of the body of the talus treated by primary Blair tibiotalar fusion. CLINICAL PRESENTATION AND INTERVENTION: A very severely comminuted open fracture of the body of the talus was treated on the same day of injury by debridement and tibiotalar fusion using the Blair fusion technique. CONCLUSION: Blair fusion may be indicated in cases of severely comminuted fractures of the talar body. It has the advantage of giving a near-normal appearance to the foot, producing less shortening and allowing motion to remain at the talonavicular and anterior subtalar joints. PMID- 12566970 TI - Asymptomatic massive hypertriglyceridemia in an octogenarian. AB - OBJECTIVE: To report the case of an 85-year-old man with asymptomatic massive hypertriglyceridemia (MHTG). CLINICAL PRESENTATION AND INTERVENTION: Our case was a non-smoker, healthy 85-year-old Caucasian male, with no excessive alcohol intake and no evidence of an excessive sedentary lifestyle, body mass index = 23.2 kg/m(2), BP = 125/85 mm Hg and plasma triglyceride (TG) >1,000 mg/dl. The MHTG was an incidental finding at the age of 70. He had no cardiovascular disease, xanthomas, xanthelasmas or keratic precipitate. During the last 15 years, his average TG plasma levels showed a significant variability independent of specific diet treatment and fibrate therapy. Liver ultrasound examination excluded hepatomegaly and fatty degeneration. Carotid artery ultrasound showed only intimal thickening in both carotid bifurcations. CONCLUSION: In this patient, MHTG had been silent for many years, with no evidence of coronary heart disease and liver fatty degeneration, both typical complications present in MHTG subjects with low high-density lipoprotein. Hence, this case must be considered as a rarity. PMID- 12566971 TI - Ethical issues in organ transplantation. AB - Clinical organ transplantation has been recognized as one of the most gripping medical advances of the century as it provides a way of giving the gift of life to patients with terminal failure of vital organs, which requires the participation of other fellow human beings and of society by donating organs from deceased or living individuals. The increasing incidence of vital organ failure and the inadequate supply of organs, especially from cadavers, has created a wide gap between organ supply and organ demand, which has resulted in very long waiting times to receive an organ as well as an increasing number of deaths while waiting. These events have raised many ethical, moral and societal issues regarding supply, the methods of organ allocation, the use of living donors as volunteers including minors. It has also led to the practice of organ sale by entrepreneurs for financial gains in some parts the world through exploitation of the poor, for the benefit of the wealthy. The current advances in immunology and tissue engineering and the use of animal organs, xenotransplantation, while offering very promising solutions to many of these problems, also raise additional ethical and medical issues which must be considered by the medical profession as well as society. This review deals with the ethical and moral issues generated by the current advances in organ transplantation, the problem of organ supply versus organ demand and the appropriate allocation of available organs. It deals with the risks and benefits of organ donation from living donors, the appropriate and acceptable methods to increase organ donation from the deceased through the adoption of the principle of 'presumed consent', the right methods of providing acceptable appreciation and compensation for the family of the deceased as well as volunteer and altruistic donors, and the duties and responsibilities of the medical profession and society to help fellow humans. The review also deals with the appropriate and ethically acceptable ways of utilizing the recent advances of stem cell transplantation from adult versus fetal donors, tissue engineering and the use of organs from animals or xenotransplantation. Data provided in support of the concept that clinical organ and tissue transplantation can be more beneficial and life saving if everyone involved in the process, including physicians and medical institutions, respect and consider the best interests of the patients, as well as honor the ethical, moral and religious values of society and are not tempted to seek personal fame or financial rewards. PMID- 12566972 TI - Roles of microtubule dynamics and small GTPase Rac in endothelial cell migration and lamellipodium formation under flow. AB - Endothelial cell (EC) migration is required for vascular development and wound healing. We investigated the roles of microtubule (MT) dynamics and the small GTPase Rac in the fluid shear stress-induced protrusion of lamellipodia and enhancement of migration of bovine aortic ECs (BAECs). Shear stress increased lamellipodial protrusion and cell migration. Treating BAECs with paclitaxel (Taxol), an MT-stabilizing agent, inhibited lamellipodial protrusion and reduced migration speed in both the static and sheared groups. After Taxol washout, both lamellipodial protrusion and cell migration increased in the flow direction. Taxol treatment also decreased the shear-induced Rac activation. Transfection of BAECs with a dominant negative mutant of Rac1 inhibited lamellipodial protrusion and cell migration under static and shear conditions. Transfection with an activated mutant of Rac1 induced lamellipodia in all directions and attenuated the shear-induced migration, suggesting that an appropriate level of Rac activity and a polarized lamellipodial protrusion are important for cell migration under static and shear conditions. Our findings suggest that MT dynamics and optimum Rac activation are required for the polarized protrusion of lamellipodia that drives the directional EC migration under flow. PMID- 12566973 TI - PGE2 action in human coronary artery smooth muscle: role of potassium channels and signaling cross-talk. AB - Cyclic AMP-stimulating agents are powerful vasodilators, but our knowledge of the signal transduction mechanisms of these agents, particularly in human arteries, is limited. We now report direct molecular effects of prostaglandin E(2) (PGE(2)) on cultured human coronary artery smooth muscle cells (HCASMC). Patch-clamp studies revealed that 10 microM PGE(2) opens a high-conductance (approximately 200 pS), calcium-stimulated potassium (BK(Ca)) channel in intact HCASMC. In contrast, PGE(2) had no direct effect on channels in cell-free patches, indicating involvement of a soluble second messenger. Enzyme immunoassay demonstrated that PGE(2) enhances production of cAMP in HCASMC, but does not increase [cGMP]. Furthermore, forskolin, CPT-cAMP, or CPT-cGMP mimicked the stimulatory effect of PGE(2) on BK(Ca) channel activity. Interestingly, the response to PGE(2) was unaffected by inhibiting the cAMP-dependent protein kinase, but was antagonized by inhibitors of the cGMP-dependent protein kinase (PKG). Furthermore, cAMP-stimulated PKG activity mimicked the effect of PGE(2). These studies suggest a novel PGE(2) action in human arteries: opening of BK(Ca) channels via cAMP cross-activation of PKG in HCASMC. It is proposed that this signaling mechanism may mediate the vasodilatory response to cAMP-dependent agents in the human coronary and other vascular beds. PMID- 12566974 TI - Nitric oxide releases calcitonin-gene-related peptide from rat dura mater encephali promoting increases in meningeal blood flow. AB - Nitric oxide (NO) and calcitonin-gene-related peptide (CGRP) are implicated in the pathophysiology of vascular headaches. We studied the interaction of these two vasodilatory mediators in an animal model and suggest that NO may increase meningeal blood flow not only by its direct vasodilatory action but also by stimulating CGRP release. First, CGRP release from the rat cranial dura mater was measured in vitro using an enzyme immunoassay. Hemisected skulls with adhering dura mater were filled with synthetic interstitial fluid and stimulated with the NO donor diethylamine-NONOate (10(-5)-10(-3) M) or with NO gas (1,000 ppm), which caused concentration-dependent increases in CGRP release up to 166.8%. Second, meningeal blood flow was recorded in vivo in the exposed dura mater using laser Doppler flowmetry. Topical application of the NO donors NONOate, S-nitroso-N acetylpenicillamine and N-ethyl-2-(1-ethyl-2-hydroxy-2-nitrosohydrazino) ethenamine (10(-5)-10(-3) M) caused concentration-dependent increases in blood flow. These increases were significantly reduced by local preliminary application of the CGRP receptor antagonist CGRP(8-37) (10(-4) M). We conclude that NO stimulates the release of CGRP from dural afferents. The blood-flow-increasing effect of NO seems to be partly mediated by CGRP. The interaction of NO and CGRP may be relevant for the development of vascular headaches. PMID- 12566975 TI - Association between the G protein beta3 subunit 825t allele and radial artery hypertrophy. AB - The GNB3 C825T gene polymorphism has recently been identified and associated with hypertension, obesity and left ventricular hypertrophy. The aim of the study was to determine the relationship between the C825T polymorphism of the gene encoding for the G protein beta3 subunit (GNB3 C825T) and vascular hypertrophy. We studied a cohort of 306 subjects (age 49 +/- 12 years) without evidence of cardiovascular disease and never treated with cardiovascular drugs. Vascular phenotypes were evaluated for the common carotid and radial arteries using high-resolution echo tracking devices. Genotype frequencies were in agreement with the Hardy-Weinberg equilibrium. For the radial artery, mean wall thickness was significantly higher in subjects carrying the 825T allele than in CC genotype subjects (240 +/- 54 microm for CT genotype and 241 +/- 53 microm for TT genotype vs. 222 +/- 52 microm for CC genotype, p = 0.01). The frequency of the 825T allele was significantly different in subjects with (52%) and without (35%) radial artery hypertrophy (chi(2) = 10.88, p < 0.001). The relative risk of radial artery hypertrophy in subjects carrying the 825T allele compared with those with the CC genotype was 3.02 (95% CI 1.53- 5.95). A logistic regression analysis indicated that the positive and significant association between the 825T allele and radial artery hypertrophy was independent of age, blood pressure, gender and BMI. In contrast, no association between genotypes and carotid artery wall thickening was observed. These results suggest that some genetic characteristics determine in part the patterns of radial artery geometrical changes. As the 825T allele is associated with vascular hypertrophy of a muscular artery but not with structural changes of an elastic artery, we hypothesize that the 825T allele may be a genetic marker of arteriolosclerosis. PMID- 12566976 TI - Effects of insulin on coronary blood flow in anesthetized pigs. AB - Insulin can influence the vasculature by a sympathetically mediated vasoconstriction and a vasodilatation; the latter effect predominates in the renal circulation of anesthetized pigs. We determined the effect of intravenous infusion of insulin on coronary blood flow in pentobarbitone-anesthetized pigs at constant heart rate, arterial pressure and blood levels of glucose and potassium. In 6 pigs, infusion of 0.004 IU kg(-1) min(-1) of insulin decreased coronary flow despite increasing left ventricular dP dT(max)(-1); when the latter was abolished by propranolol, the coronary flow response was augmented. The mechanisms of this response were examined in 22 pigs given propranolol. Phentolamine changed coronary flow response to an increase (6 pigs) and this was abolished by intracoronary injection of N(omega)-nitro-L-arginine methyl ester (L-NAME; 5 pigs). L-NAME augmented coronary flow response (6 pigs) and this was abolished by phentolamine (5 pigs). In 18 pigs given propranolol, three incremental doses of insulin caused graded coronary flow decreases whether L-NAME was given (6 pigs) or not (6 pigs) beforehand, and caused graded coronary flow increases after phentolamine (6 pigs). Thus insulin caused a coronary vasoconstriction mediated by sympathetic alpha-adrenergic effects and a vasodilatation related to the release of nitric oxide. The net effect was a coronary vasoconstriction. PMID- 12566977 TI - Atherosclerotic expansive remodeled plaques: a wolf in sheep's clothing. AB - Geometric arterial remodeling is an important determinant of luminal narrowing in atherosclerotic disease. Expansive remodeling retards while constrictive remodeling accelerates luminal narrowing by plaque formation. Cross-sectional as well as follow-up studies revealed that expansive remodeling is associated with adverse cardiovascular events and a vulnerable plaque phenotype. Although the relation between expansive remodeling and plaque vulnerability is associative rather than causal, expansively remodeled plaques should be considered as a wolf in sheep's clothes. Further understanding of the processes that regulate arterial remodeling and plaque rupture may lead to new strategies to responsibly manipulate these processes for the benefit of patient outcomes. PMID- 12566978 TI - Perivascular endothelial implants inhibit intimal hyperplasia in a model of arteriovenous fistulae: a safety and efficacy study in the pig. AB - Vascular access complications are a major problem in hemodialysis patients. Native arteriovenous fistulae, historically the preferred mode of access, have a patency rate of only 60% at 1 year. The most common mode of failure is due to progressive stenosis at the anastomotic site. We have previously demonstrated that perivascular endothelial cell implants inhibit intimal thickening following acute balloon injury in pigs and now seek to determine if these implants provide a similar benefit in the chronic and more complex injury model of arteriovenous anastomoses. Side-to-side femoral artery-femoral vein anastomoses were created in 24 domestic swine and the toxicological, biological and immunological responses to allogeneic endothelial cell implants were investigated 3 days and 1 and 2 months postoperatively. The anastomoses were wrapped with polymer matrices containing confluent porcine aortic endothelial cells (PAE; n = 14) or control matrices without cells (n = 10). PAE implants significantly reduced intimal hyperplasia at the anastomotic sites compared to controls by 68% (p < 0.05) at 2 months. The beneficial effects of the PAE implants were not due to differences in the rates of reendothelialization between the groups. No significant immunological response to the allogeneic endothelial cells that impacted on efficacy was detected in any of the pigs. No apparent toxicity was observed in any of the animals treated with endothelial implants. These data suggest that perivascular endothelial cell implants are safe and reduce early intimal hyperplasia in a porcine model of arteriovenous anastomoses. PMID- 12566979 TI - Real-time detection of activation patterns in individual platelets during thromboembolism in vivo: differences between thrombus growth and embolus formation. AB - Knowledge on single platelet behavior and intracellular mechanisms during thromboembolism in vivo is scarce. In the present study, we used a new method that enables real-time detection and quantification of activation of individual platelets participating in a thromboembolic process in vivo, using their intracellular free Ca(2+) concentration ([Ca(2+)](i)) as a marker of activation. Isolated platelets were labeled with the Ca(2+)-sensitive fluorescent probe fluo 3 and injected into anesthetized rabbits so that 0.5-1% of their circulating platelets were labeled. Wall puncture of mesenteric arterioles resulted in thrombus formation followed by embolization. Fluorescence intensity changes of labeled platelets participating in this process were quantified. Within 30 min after injection, labeled platelets behaved similarly to native platelets, and fluorescence intensity was not influenced by dye leakage. Upon adherence to the stationary thrombus, platelets exhibited a prolonged [Ca(2+)](i) increase, accompanied by shape change and degranulation, which is consistent with a role for strong platelet agonists like collagen. In contrast, when platelets adhered to a growing embolus their [Ca(2+)](i) rise was transient, and they hardly showed shape change and degranulation, suggesting the involvement of weaker agonists like ADP. These results show, for the first time, the relation between single platelet activation patterns, which are different during thrombus growth and embolus formation, and their behavior in a thromboembolic process in vivo. PMID- 12566982 TI - Marginal division of the neostriatum: a subcortical memory center. AB - The marginal division (MrD) is a pan-shaped subdivision in the caudal margin of the neostriatum newly discovered in the brains of the rat, cat, monkey and humans. A variety of intensely expressed neuropeptides and monoamines and their receptors were identified in the fibers, terminals and neuronal somata in the MrD with immunohistochemical and patch clamp methods. The MrD was shown to be involved in learning and memory by double-blind studies of Y-maze learning and long-term potentiation in rats. c-Fos expression and tract-tracing techniques with immunoelectronmicroscopy indicated that the MrD is a new component of the limbic system and is a key linking area between the limbic system and the basal nucleus of Meynert. Functional magnetic resonance image (fMRI) studies illustrated that the MrD and the prefrontal cortex are involved in digital working memory in the human brain. A cerebral hemorrhage case report confirmed the findings with fMRI. In conclusion, based on the position of the MrD, its advanced development in higher mammalian brains, abundant blood supply and diverse connections with other memory-related structures, MrD is likely to be an important subcortical center of learning and memory. PMID- 12566983 TI - Membrane glycine transport proteins. AB - Structurally, the simplest amino acid is glycine, and it has a number of important yet distinct functions in the body. This review focuses on the different transport systems and the associated carrier proteins for glycine that are responsible for its movement across biological membranes. Transport proteins in the class GLYT appear to be the most specific for glycine. However, the B0+ system also carries significant amounts of glycine. Other amino acid transport systems capable of carrying small amounts of glycine are ASC, asc and system L. In addition, an ATP-dependent transport process exists that takes up glycine into synaptic vesicles at nerve endings. This is known as the vesicular inhibitory amino acid transporter since, in addition to glycine, it can transport possibly two other inhibitory neurotransmitters. PMID- 12566981 TI - Ischemia-reperfusion-related repair deficit after oxidative stress: implications of faulty transcripts in neuronal sensitivity after brain injury. AB - Diseases of the heart are the No. 1 killer in industrialized countries. Brain injury can develop as a result of cerebral ischemia-reperfusion due to stroke (brain attack) and other cardiovascular diseases. Learning about the disease is the best way to reduce disability and death. We present here whether gene repair activities are associated with neuronal death in an ischemia-reperfusion model that simulates stroke in male Long-Evans rats. This experimental stroke model is known to induce necrosis in the ischemic cortex. Cerebral ischemia causes overactivation of membrane receptors and accumulation of extracellur glutamate and intracellular calcium, which activates neuronal nitric oxide synthase, causing damage to lipids, proteins, and nucleic acids, and reduces energy sources with consequent functional deterioration, leading to cell death. Restoration processes normally repair genes with few errors. However, ischemia elevates oxidative DNA lesions despite these repair mechanisms. These episodes concurrently occur with the induction of immediate-early genes that critically activate other late genes in the signal transduction pathway. Damage, repair, and transcription of the c-FOS gene are presented here as examples, because Fos peptide, one of the components of activator protein 1, activates nerve growth factor and repair mechanisms. The results of our studies show that treatments with 7-nitroindazole, a specific inhibitor of nitric oxide synthase known to attenuate nitric oxide, oxidative DNA lesions, and necrosis, increase intact c fos mRNA levels after stroke. This suggests that the accuracy of gene expression could be accounted for the recovery of cellular function after cerebral injury. PMID- 12566984 TI - Cancer immunogene therapy: a review. AB - Although immunotherapy has long held out promise as a specific, potent approach to cancer therapy, clinical applications have been unrewarding to date. However, advances in gene transfer technology and basic immunology have opened new avenues to stimulate antitumor immune responses including immunogene therapy. Many different approaches to immunogene therapy have been identified. These include transferring genes encoding proinflammatory proteins to tumor cells, suppressing immunosuppressive gene expression, and transferring proinflammatory genes and/or tumor antigen genes to professional antigen-presenting cells. In some cases, genes are transferred to tumor or antigen-presenting cells in situ. In others, gene transfer is performed ex vivo as part of preparing an anticancer vaccine. We discuss the underlying approach, relative success, and clinical application of various cancer immunogene therapy strategies, paying particular attention to immunogene therapy vaccines. Large numbers of preclinical studies have been reported, but only scattered clinical trial results have appeared in the literature. Although very successful preclinically, the ideal cancer immunogene therapy approach remains to be determined and will likely vary with tumor type. Clinical impact may be improved in the future as treatment protocols are refined. PMID- 12566985 TI - The apoptotic action of the retinoid CD437/AHPN: diverse effects, common basis. AB - Retinoids, such as all-TRANS-retinoic acid (RA), have found applications in several different types of (cancer) therapies. The synthetic retinoid 6-[3-(1 adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (CD437 or AHPN), an RA receptor (RAR)gamma agonist, not only induces RARgamma-dependent differentiation, but in contrast to RA, it also induces RARgamma-independent apoptosis in many tumor cells. This observation makes this and similar new retinoids very interesting from a clinical perspective. Several genes have been associated with CD437/AHPN-mediated apoptosis, but the multiple activities of this compound and the apparent cell-type-specific responses to treatment have made it difficult to discern a common biochemical basis for the mechanism of its apoptotic action. In this brief review, we present a model which links all CD437/AHPN-associated apoptotic effects. CD437/AHPN rapidly induces DNA adduct formation through an as yet unknown reaction which is independent of cell type. This is followed by a cell-type-specific, largely p53-independent DNA damage response which can result in engagement of multiple cell death pathways and activation of caspases as a common endpoint. At the same time, the RARgamma-dependent pathway leads to regulation of differentiation-associated, cell-type-specific genes. CD437/AHPN, with its simultaneous differentiation and apoptosis-inducing activities, is a good prototype for new drugs which may be clinically more efficacious than those with a single activity. PMID- 12566986 TI - The effects of spironolactone monotherapy on blood pressure and myocardial remodeling in spontaneously hypertensive rats: a stereological study. AB - The aim of this study was to evaluate the cardiac structure of spontaneously hypertensive rats (SHRs) treated with different doses of spironolactone. Twenty SHRs were separated into four groups and treated for 13 weeks, as follows: one control group and three spironolactone treatment groups receiving doses of 5, 10 or 30 mg/kg/day. The spironolactone treatment either attenuated or prevented the tendency for increased blood pressure. However, the myocardial structure was not significantly affected by the spironolactone monotherapy treatment (all doses); it showed hypertrophied cardiac myocytes, focal areas of reactive fibrosis, inflammatory infiltrate and a decrease in the density of intramyocardial microvessels. None of the cardiac myocyte stereological parameters in the left ventricular myocardium showed significant differences among the SHR groups. The cardiac myocyte volume density was around 80%, the cardiac myocyte surface density varied from 3.6 to 4.1 x 10(4) mm2/mm3 and the cardiac myocyte mean cross sectional area varied from 351 to 415 micro m2. The connective tissue volume density of the SHRs treated with the highest dose of spironolactone was 75% lower than in the control SHRs, and this was the only significant difference found for this parameter among SHR groups. The intramyocardial vessels showed some differences when the control SHRs and the other SHRs were compared. The lowest intramyocardial vessel volume density was found in the control group (more than 20% lower than that in the treated SHRs), but no significant difference was detected among the treated SHRs (all doses). The intramyocardial vessel length density (Lv[v]) and surface density (Sv[v]) showed a similar tendency, being significantly greater in the treated SHRs than in the control rats. The Lv[v] was 45% greater in the high-dose spironolactone group than in the control group, and it was 28% greater in the high-dose spironolactone SHRs than in the other treated SHRs. The Sv[v] was 50% greater in the high-dose spironolactone SHRs than in both control and low-dose spironolactone SHRs. Long-term spironolactone monotherapy showed a partial effect in the preservation of intramyocardial vessels and also in the attenuation of interstitial fibrosis. PMID- 12566987 TI - Nitric oxide mediates lung injury induced by ischemia-reperfusion in rats. AB - Nitric oxide (NO) has been reported to play a role in lung injury (LI) induced by ischemia-reperfusion (I/R). However, controversy exists as to the potential beneficial or detrimental effect of NO. In the present study, an in situ, perfused rat lung model was used to study the possible role of NO in the LI induced by I/R. The filtration coefficient (Kfc), lung weight gain (LWG), protein concentration in the bronchoalveolar lavage (PCBAL), and pulmonary arterial pressure (PAP) were measured to evaluate the degree of pulmonary hypertension and LI. I/R resulted in increased Kfc, LWG, and PCBAL. These changes were exacerbated by inhalation of NO (20-30 ppm) or 4 mM L-arginine, an NO precursor. The permeability increase and LI caused by I/R could be blocked by exposure to 5 mM N omega-nitro-L-arginine methyl ester (L-NAME; a nonspecific NO synthase inhibitor), and this protective effect of L-NAME was reversed with NO inhalation. Inhaled NO prevented the increase in PAP caused by I/R, while L-arginine had no such effect. L-NAME tended to diminish the I/R-induced elevation in PAP, but the suppression was not statistically significant when compared to the values in the I/R group. These results indicate that I/R increases Kfc and promotes alveolar edema by stimulating endogenous NO synthesis. Exogenous NO, either generated from L-arginine or delivered into the airway, is apparently also injurious to the lung following I/R. PMID- 12566988 TI - Effects of the calcium channel blocker amlodipine on serum and aortic cholesterol, lipid peroxidation, antioxidant status and aortic histology in cholesterol-fed rabbits. AB - Reactive oxygen metabolites and oxidized fatty acids are proinflammatory and are involved in the pathophysiology of atherosclerosis. Amlodipine, a unique third generation dihydropyridine-type calcium channel blocker, seems to exert atheroprotective effects through its antioxidant properties related to its chemical structure and independent of its calcium channel-blocking effect. In this study, the interactions of amlodipine with major cellular antioxidants were investigated in order to elucidate the mechanisms underlying its atheroprotective effects. New Zealand white male rabbits were fed regular chow (group 1), chow with 1% cholesterol (group 2), regular chow plus 5 mg/kg/day amlodipine per os (group 3) and 1% cholesterol plus amlodipine (group 4) for 8 weeks. Total cholesterol, malondialdehyde (MDA) and vitamin E concentrations and catalase and superoxide dismutase (SOD) activities were determined in blood drawn before and after the experimental period. Aortic tissue was examined for atherosclerotic changes and aortic total cholesterol, MDA, catalase and SOD were determined. At the end of the 8-week treatment period, serum total cholesterol and plasma MDA were elevated in groups 2 and 4. In group 2, serum vitamin E and plasma SOD diminished (p < 0.05) and catalase increased (p < 0.05). In group 4, SOD activity increased at the end of treatment. MDA levels were lower and plasma SOD activities were higher in group 4 than in group 2. Aortic tissue investigations revealed higher total cholesterol and MDA concentrations and catalase activities in group 2 than in group 4, and the highest tissue SOD activity was recorded in group 4 (p < 0.05 for all comparisons). Morphological examination of aortic tissues exhibited endothelial disarrangement and lipid deposition in group 2. Histopathological alterations related to atherogenesis were less in group 4 than in group 2. Amlodipine seems to exert atheroprotective effects by reducing aortic cholesterol accumulation and blood and aortic lipid peroxidation, enhancing SOD activity both in blood and aortic tissue and suppressing the consumption of vitamin E. On the other hand, the suppression of catalase activity in blood and the aorta interferes with the drug's well-known antioxidant effects. PMID- 12566989 TI - Inhibition of established subcutaneous and metastatic murine tumors by intramuscular electroporation of the interleukin-12 gene. AB - In vivo electroporation (EP) of the murine interleukin-12 (IL-12) gene in an expression plasmid (pIL-12) was evaluated for antitumor activity. EP transfer of pIL-12 into mouse quadriceps muscles elicited significant levels of serum IL-12 and interferon-gamma. Intramuscular EP of pIL-12 resulted in complete regression or substantial inhibition of 38C13 B-cell lymphoma, whereas pIL-12 delivered by gene gun or intramuscular injection without EP showed little therapeutic effect. Impressive antitumor activity by intramuscular EP was also demonstrated in animals with advanced malignant disease. At day 14 after 38C13 tumor inoculation, all animals were found to carry large tumors and to have metastases; without treatment, most died within a week. A single intramuscular EP of pIL-12 resulted in regression of 50% of large subcutaneous tumors and significantly prolonged the lifespan of these animals. Moreover, animals that were previously cured of 38C13 tumors by in vivo EP treatment significantly suppressed tumor growth when challenged 60 days later. In vivo EP of the IL-12 gene was also effective in suppressing subcutaneous and lung metastatic tumors of CT-26 colon adenocarcinoma and B16F1 melanoma cells. Together, these results show that intramuscular electrotransfer of the IL-12 gene may represent a simple and effective strategy for cancer treatment. PMID- 12566990 TI - Characterization of reduced expression of glycine N-methyltransferase in cancerous hepatic tissues using two newly developed monoclonal antibodies. AB - Glycine N-methyltransferase (GNMT) is a protein with multiple functions. Recently, two Italian siblings who had hepatomegaly and chronic elevation of serum transaminases were diagnosed to have GNMT deficiency caused by inherited compound heterozygosity of the GNMT gene with missence mutations. To evaluate the expression of GNMT in cell lines and tissues from hepatocellular carcinoma (HCC) patients, we produced two monoclonal antibodies (mAbs) 4-17 and 14-1 using two recombinant GNMT fusion proteins. M13 phage peptide display showed that the reactive epitopes of mAbs 4-17 and 14-1 were amino acid residues 11-15 and 272 276 of human GNMT, respectively. The dissociation constants of the binding between GNMT and mAbs were 1.7 x 10(-8) M for mAb 4-17 and 1.8 x 10(-9) M for mAb 14-1. Both mAbs can identify GNMT present in normal human and mouse liver tissues using Western blotting (WB) and immunohistochemical staining assay (IHC). In addition, WB with both mAbs showed that none of 2 hepatoblastoma and 5 HCC cell lines expressed GNMT. IHC demonstrated that 50% (13/26) of nontumorous liver tissues and 96% (24/25) of HCC tissues did not express GNMT. Therefore, the expression of GNMT was downregulated in human HCC. PMID- 12566991 TI - Genomic structure, gene expression, and promoter analysis of human multidrug resistance-associated protein 7. AB - The multidrug resistance-associated protein (MRP) subfamily transporters associated with anticancer drug efflux are attributed to the multidrug-resistance of cancer cells. The genomic organization of human multidrug resistance associated protein 7 (MRP7) was identified. The human MRP7 gene, consisting of 22 exons and 21 introns, greatly differs from other members of the human MRP subfamily. A splicing variant of human MRP7, MRP7A, expressed in most human tissues, was also characterized. The 1.93-kb promoter region of MRP7 was isolated and shown to support luciferase activity at a level 4- to 5-fold greater than that of the SV40 promoter. Basal MRP7 gene expression was regulated by 2 regions in the 5'-flanking region at -1,780-1,287 bp, and at -611 to -208 bp. In Madin Darby canine kidney (MDCK) cells, MRP7 promoter activity was increased by 226% by genotoxic 2-acetylaminofluorene and 347% by the histone deacetylase inhibitor, trichostatin A. The protein was expressed in the membrane fraction of transfected MDCK cells. PMID- 12566992 TI - Purification and characterization of a novel isoallergen of a major Bermuda grass pollen allergen, Cyn d 1. AB - A novel immunoreactive isoallergen of a major Bermuda grass pollen allergen, Cyn d 1, was purified by the use of a combination of various chromatographic techniques, including high-performance liquid chromatography. This new isoallergen has a pI value of 9.1 and shows significant N-terminal sequence homology with other isoforms. Carbohydrate composition analysis revealed a 10.4% carbohydrate content consisting of 7 different sugar moieties, including arabinose, fucose, galactose, glucose, mannose, xylose and N-acetylglucosamine, as well as a trace amount of rhamnose. Upon periodate oxidation, the binding activities of the Cyn d 1 isoform to murine monoclonal antibodies and human serum IgE and IgG were reduced, suggesting the importance of the carbohydrate moiety in the immune response. The availability of the purified Cyn d 1 basic isoform will allow for further structural and immunological characterization, and ultimately for the design of an appropriate therapy. PMID- 12566993 TI - Activation of Th1 immunity is a common immune mechanism for the successful treatment of hepatitis B and C: tetramer assay and therapeutic implications. AB - Both chronic hepatitis B and C virus (HBV and HCV) infections respond ineffectively to current antiviral therapies. Recent studies have suggested that treatment outcomes may depend on the development of type 1 T helper (Th1) and Th2 cell responses. Specifically, activation of Th1 immunity may play a major role in successfully treating hepatitis B and C. This model was revisited herein by evaluating immune responses in 36 HBV and 40 HCV patients with or without treatment, in an attempt to find a common immune mechanism for successful treatment. The immune responses in all examined cases were studied by peripheral blood mononuclear cell (PBMC) proliferation and cytokine responses to viral antigens, cytotoxic T lymphocyte (CTL) responses, enzyme-linked immunospot (ELISPOT) assay, and tetramer staining of virus-specific CD8+ T cells. The overall results revealed that all responders among both HBV- and HCV-infected cases displayed significantly higher PBMC proliferation to viral antigens with a predominant Th1 cytokine profile. Furthermore, the Th1-dominant responses were associated with significant enhancement of CTL activities and were correlated with ELISPOT data, while non-responders responded more weakly. During therapy, the numbers of tetramer-staining, virus-specific CD8+ T cells showed greater increases in responders than in non-responders (p = 0.001). The frequencies determined by the tetramer assay were approximately 200-fold higher than data estimated by limiting-dilution analysis. In conclusion, activation of Th1 immunity accompanied by enhancement of CTL activity during therapy is a common immune mechanism for successfully treating hepatitis B and C, and therefore may have important therapeutic implications. PMID- 12566994 TI - The virotoxin model of HIV-1 enteropathy: involvement of GPR15/Bob and galactosylceramide in the cytopathic effects induced by HIV-1 gp120 in the HT-29 D4 intestinal cell line. AB - BACKGROUND: Malabsorption and diarrhea are common, serious problems in AIDS patients, and are in part due to the incompletely understood entity HIV enteropathy. Our prior in vitro work has shown that increased transepithelial permeability and glucose malabsorption, similar to HIV enteropathy, are caused by HIV surface protein gp120, although the mechanism remains unclear. RESULTS: We studied the effects of HIV surface protein gp120 on the differentiated intestinal cell line HT-29-D4, specifically the effects on microtubules, transepithelial resistance, and sodium glucose cotransport. gp120 induced extensive microtubule depolymerization, an 80% decrease in transepithelial resistance, and a 70% decrease in sodium-dependent glucose transport, changes closely paralleling those of HIV enteropathy. The effects on transepithelial resistance were used to study potential inhibitors. Neutralizing antibodies to GPR15/Bob but not to CXCR4 (the coreceptor allowing infection with these HIV strains) inhibited these effects. Antibodies to galactosylceramide (GalCer) and a synthetic analog of GalCer also inhibited the gp120-induced changes, suggesting the involvement of GalCer enriched lipid rafts in gp120 binding to intestinal epithelial cells. CONCLUSION: We conclude that direct HIV infection and gp120-induced cytopathic effects are distinct phenomena. While in vivo confirmation is needed to prove this, gp120 could be a virotoxin significantly contributing to HIV enteropathy. PMID- 12566995 TI - MONTE: An automated Monte Carlo based approach to nuclear magnetic resonance assignment of proteins. AB - A general-purpose Monte Carlo assignment program has been developed to aid in the assignment of NMR resonances from proteins. By virtue of its flexible data requirements the program is capable of obtaining assignments of both heavily deuterated and fully protonated proteins. A wide variety of source data, such as inter-residue scalar connectivity, inter-residue dipolar (NOE) connectivity, and residue specific information, can be utilized in the assignment process. The program can also use known assignments from one form of a protein to facilitate the assignment of another form of the protein. This attribute is useful for assigning protein-ligand complexes when the assignments of the unliganded protein are known. The program can be also be used as an interactive research tool to assist in the choice of additional experimental data to facilitate completion of assignments. The assignment of a deuterated 45 kDa homodimeric Glutathione-S transferase illustrates the principal features of the program. PMID- 12566996 TI - Data requirements for reliable chemical shift assignments in deuterated proteins. AB - The information required for chemical shift assignments in large deuterated proteins was investigated using a Monte Carlo approach (Hitchens et al., 2002). In particular, the consequences of missing amide resonances on the reliability of assignments derived from C alpha and CO or from C alpha and C beta chemical shifts was investigated. Missing amide resonances reduce both the number of correct assignments as well as the confidence in these assignments. More significantly, a number of undetectable errors can arise when as few as 9% of the amide resonances are missing from the spectra. However, the use of information from residue specific labeling as well as local and long-range distance constraints improves the reliability and extent of assignment. It is also shown that missing residues have only a minor effect on the assignment of protein ligand complexes using C alpha and CO chemical shifts and C alpha inter-residue connectivity, provided that the known chemical shifts of the unliganded protein are utilized in the assignment process. PMID- 12566997 TI - The use of model selection in the model-free analysis of protein dynamics. AB - Model-free analysis of NMR relaxation data, which is widely used for the study of protein dynamics, consists of the separation of the global rotational diffusion from internal motions relative to the diffusion frame and the description of these internal motions by amplitude and timescale. Five model-free models exist, each of which describes a different type of motion. Model-free analysis requires the selection of the model which best describes the dynamics of the NH bond. It will be demonstrated that the model selection technique currently used has two significant flaws, under-fitting, and not selecting a model when one ought to be selected. Under-fitting breaks the principle of parsimony causing bias in the final model-free results, visible as an overestimation of S2 and an underestimation of taue and Rex. As a consequence the protein falsely appears to be more rigid than it actually is. Model selection has been extensively developed in other fields. The techniques known as Akaike's Information Criteria (AIC), small sample size corrected AIC (AICc), Bayesian Information Criteria (BIC), bootstrap methods, and cross-validation will be compared to the currently used technique. To analyse the variety of techniques, synthetic noisy data covering all model-free motions was created. The data consists of two types of three dimensional grid, the Rex grids covering single motions with chemical exchange [S2,taue,Rex], and the Double Motion grids covering two internal motions [S f 2,S s 2,tau s ]. The conclusion of the comparison is that for accurate model-free results, AIC model selection is essential. As the method neither under, nor over fits, AIC is the best tool for applying Occam's razor and has the additional benefits of simplifying and speeding up model-free analysis. PMID- 12566998 TI - Prediction algorithm for amino acid types with their secondary structure in proteins (PLATON) using chemical shifts. AB - The algorithm PLATON is able to assign sets of chemical shifts derived from a single residue to amino acid types with its secondary structure (amino acid species). A subsequent ranking procedure using optionally two different penalty functions yields predictions for possible amino acid species for the given set of chemical shifts. This was demonstrated in the case of the alpha-spectrin SH3 domain and applied to 9 further protein data sets taken from the BioMagRes database. A database consisting of reference chemical shift patterns (reference CSPs) was generated from assigned chemical shifts of proteins with known 3D structure. This reference CSP database is used in our approach for extracting distributions of amino acid types with their most likely secondary structure elements (namely alpha-helix, beta-sheet, and coil) for single amino acids by comparison with query CSPs. Results obtained for the 10 investigated proteins indicates that the percentage of correct amino acid species in the first three positions in the ranking list, ranges from 71.4% to 93.2% for the more favorable penalty function. Where only the top result of the ranking list for these 10 proteins is considered, 36.5% to 83.1% of the amino acid species are correctly predicted. The main advantage of our approach, over other methods that rely on average chemical shift values is the ability to increase database content by incorporating newly derived CSPs, and therefore to improve PLATON's performance over time. PMID- 12566999 TI - Myristoylation as a general method for immobilization and alignment of soluble proteins for solid-state NMR structural studies. AB - N-terminal myristoylation of the immunoglobulin-binding domain of protein G (GB1) from group G Streptococcus provides the means to bind the protein to aligned phospholipid bilayers for solid-state NMR structural studies. The myristoylated protein is immobilized by its interactions with bilayers, and the sample alignment enables orientationally dependent 15N chemical shifts and 1H-15N dipolar couplings to be measured. Spectra calculated for the average solution NMR structure of the protein at various orientations with respect to the magnetic field direction were compared to the experimental spectrum. The best fit identified the orientation of the myristoylated protein on the lipid bilayers, and demonstrated that the protein adopts a similar structure in both its myristoylated and non-myristoylated forms, and that the structure is not grossly distorted by its interaction with the phosholipid bilayer surface or by its location in the restricted aqueous space between bilayer leaflets. The protein is oriented such that its charged sides face the phosphatidylcholine headgroups of the lipids with the single amphiphilic helix running parallel to the bilayer surface. PMID- 12567000 TI - Determination of protein global folds using backbone residual dipolar coupling and long-range NOE restraints. AB - We report the determination of the global fold of human ubiquitin using protein backbone NMR residual dipolar coupling and long-range nuclear Overhauser effect (NOE) data as conformational restraints. Specifically, by use of a maximum of three backbone residual dipolar couplings per residue (Ni-H N i, Ni-C'(i-1), H N i - C'(i-1)) in two tensor frames and only backbone H N -H N NOEs, a global fold of ubiquitin can be derived with a backbone root-mean-square deviation of 1.4 A with respect to the crystal structure. This degree of accuracy is more than adequate for use in databases of structural motifs, and suggests a general approach for the determination of protein global folds using conformational restraints derived only from backbone atoms. PMID- 12567001 TI - Evaluation of the influence of anisotropic indirect nuclear spin-spin coupling tensors on effective residual dipolar couplings for model peptides. AB - Residual dipolar couplings (RDCs) observed between nuclear spins in molecules in partially oriented media have become a valuable source of information for NMR spectroscopists seeking to structurally characterize biological macromolecules. Examination of the form of the direct (D) and indirect (J) nuclear spin-spin coupling Hamiltonians indicates that all observed RDCs contain an unknown contribution from the anisotropic part of J (Delta J) in addition to the direct dipolar contribution, DPQ. Here, we evaluate the influence of DeltaJ on RDCs through a series of DFT calculations on model peptides. Very small corrections to one-bond RDCs measured between heavy atoms in peptides and proteins are recommended: +0.51% for N-C' spin pairs, and +0.45% for C alpha-C' spin pairs. The corrections to RDCs involving at least one proton are negligible. This latter point is likely to be equally applicable to nucleic acids and oligosaccharides in addition to peptides and proteins. Finally, the orientations of the J(N, C') and J(C alpha, C') tensors in the molecular framework are reported for glycylglycine. PMID- 12567002 TI - 1H, 13C and 15N resonance assignments of the catalytic domain of human MAPK phosphatase, PAC-1. PMID- 12567003 TI - Assignment of 1H, 13C and 15N resonances of the N-terminal microtubule-binding domain of human doublecortin. PMID- 12567004 TI - Resonance assignment and topology of a clostridial repetitive oligopeptide (CROP) region of toxin A from Clostridium difficile. PMID- 12567005 TI - Phylogenetic relationships of the Dactylogyridae Bychowsky, 1933 (Monogenea: Dactylogyridea): the need for the systematic revision of the Ancyrocephalinae Bychowsky, 1937. AB - Phylogenetic analyses based on partial 18S rDNA sequences of polyonchoinean monogeneans were conducted in order to investigate the relationships between selected families and subfamilies of the Dactylogyrinea, mainly within the Dactylogyridae. We tested the status of the Ancyrocephalidae sensu Bychowsky & Nagibina (1978) and the Ancyrocephalinae sensu Kritsky & Boeger (1989). Within the Dactylogyrinea, the Diplectanidae and Dactylogyridae are well supported by maximum likelihood and maximum parsimony analyses, but their phylogenetic relationship with the Pseudomurraytrematidae remains unresolved. Phylogenetic relationships between the Pseudodactylogyrinae, Ancyrocephalinae, Ancylodiscoidinae and Dactylogyrinae indicate paraphyly of the Ancyrocephalidae sensu Bychowsky & Nagibina (1978). The group of species recently considered as the Dactylogyridae sensu Kritsky & Boeger (1989) comprises two sister groups. The first group includes the freshwater Ancyrocephalinae and the Ancylodiscoidinae. The second group includes the Pseudodactylogyrinae, Dactylogyrinae and the Ancyrocephalinae from the fish species Siganus doliatus and Tetraodon fluviatilis. The non-monophyly of the Ancyrocephalinae (i.e. the non-monophyly of the group of species recently considered as members of Ancyrocephalinae), previously suggested by Kritsky & Boeger (1989) using the morphological characters, indicates that classification of the Dactylogyridae needs to be revised. PMID- 12567006 TI - Genetic evidence for two sibling species within Contracaecum ogmorhini Johnston & Mawson, 1941 (Nematoda: Anisakidae) from otariid seals of boreal and austral regions. AB - Genetic variation of Contracaecum ogmorhini (sensu lato) populations from different otariid seals of the northern and southern hemisphere was studied on the basis of 18 enzyme loci as well as preliminary sequence analysis of the mitochondrial cyt b gene (260 bp). Samples were collected from Zalophus californianus in the boreal region and from Arctocephalus pusillus pusillus, A. pusillus doriferus and A. australis from the austral region. Marked genetic heterogeneity was found between C. ogmorhini (sensu lato) samples from the boreal and austral region, respectively. Two loci (Mdh-2 and NADHdh) showed fixed differences and a further three loci (Iddh, Mdh-1 and 6Pgdh) were highly differentiated between boreal and austral samples. Their average genetic distance was D(Nei) = 0.36 at isozyme level. At mitochondrial DNA level, an average proportion of nucleotide substitution of 3.7% was observed. These findings support the existence of two distinct sibling species, for which the names C. ogmorhini (sensu stricto) and C. margolisi n. sp., respectively, for the austral and boreal taxon, are proposed. A description for C. margolisi n. sp. is provided. No diagnostic morphological characters have so far been detected; on the other hand, two enzyme loci, Mdh-2 and NADHdh, fully diagnostic between the two species, can be used for the routine identification of males, females and larval stages. Mirounga leonina was found to host C. ogmorhini (s.s.) in mixed infections with C. osculatum (s.l.) (of which C. ogmorhini (s.l.) was in the past considered to be a synonym) and C. miroungae; no hybrid genotypes were found, confirming the reproductive isolation of these three anisakid species. The hosts and geographical range so far recorded for C. margolisi n. sp. and C. ogmorhini (s.s.) are given. PMID- 12567007 TI - Dactylogyridae (Monogenea) from the gills of Iheringichthys labrosus (Osteichthyes: Pimelodidae) from the upper Parana River floodplain, Brazil, with the proposal of Pseudovancleaveus n. g. AB - Two new species of Demidospermus Suriano, 1983 and a new genus, Pseudovancleaveus, and species are described from the gills of Iheringichthys labrosus (Pimelodidae). The fishes were collected from the upper Parana River floodplain, Brazil. D. mandi n. sp. is characterised by the distal region of the copulatoty organ having an expanded bulb and D. labrosi n. sp. by the copulatory organ having a funnel-shaped proximal extremity. The latter species also has an accessory piece comprising a variable sheath enclosing the distal shaft of the copulatory organ and two anterolateral structures resembling irregular spheres. Pseudovancleaveus paranaensis. n. g., n. sp. is characterised by a sinistral vagina, overlapping gonads, a copulatory ligament, anchors without a fold on the base and hooks with slightly inflated shanks. A new combination, Pseudovancleaveus platensis, is proposed for Vancleaveus platensis Suriano & Incorvaia, 1995. PMID- 12567008 TI - Bothriocephalus gadellus n. sp. (Cestoda: Bothriocephalidae) from the beardless codling Gadella imberbis (Vaillant) (Moridae) in the southwestern Gulf of Mexico, with a review of species of Bothriocephalus Rudolphi, 1808 reported from gadiform fishes. AB - Bothriocephalus gadellus n. sp. is described from the intestine of the beardless codling Gadella imberbis (Gadiformes: Moridae) from the southwestern Gulf of Mexico. It resembles B. scorpii, the type-species of Bothriocephalus, and B. manubriformis, the only species of the genus reported from the Gulf of Mexico. B. gadellus n. sp. differs from both species in having four excretory canals on each side of the proglottid (three per side in B. scorpii; two per side anastomosing to form an extensive plexus throughout the medulla in B. manubriformis ), 24-33 testes per proglottid (30-60 in B. scorpii; 60-100 in B. manubriformis), a total length of 31-47 mm (32-950 mm in B. scorpii; 130-1,000 mm in B. manubriformis), and a seminal receptacle and wing-like expansions on each proglottid (absent in B. scorpii and B. manubriformis), as well as lacking two sets of reproductive organs per proglottid (present in B. scorpii) and a vagina with a bulbous sphincter near the opening (present in B. manubriformis). An amendment to the generic diagnosis of Bothriocephalus might be warranted if other species of this genus are found to possess a seminal receptacle. B. gadellus n. sp. appears to be like those species of Bothriocephalus that can utilise two intermediate hosts (copepod and fish). B. ellipticus and B. neglectus are designated as species inquirendae. This is the first report of a cestode from a morid fish in the Gulf of Mexico, and the first report of a parasite from a species of Gadellus. PMID- 12567009 TI - A new notodelphyid copepod, Paranotodelphys illgi n. sp. (Copepoda: Cyclopoida), parasitic in the ascidian Corynascidia herdmani Ritter in the North Pacific. AB - A new species of the Notodelphyidae belonging to Paranotodelphys Schellenberg, 1922 is described from both sexes. The new species, Paranotodelphys illgi, can be distinguished from its congeners by the combination of its 10-segmented female antennule, the bisetose fifth legs and the extreme elongation of the caudal rami of the female, which are more than 10 times longer than wide and 4.4 times longer than the anal somite. The male has non-geniculate antennules. The new species was collected at a depth of 508m in the North Pacific Ocean, from the ascidian Corynascidia herdmani Ritter. PMID- 12567010 TI - What is Paranoplocephala macrocephala (Douthitt, 1915) (Cestoda: Anoplocephalidae)? AB - We present a redescription for Paranoplocephala macrocephala (Douthitt, 1915) in the type host, Geomys bursarius (Shaw) (hereafter PMG), and compare it with a P. macrocephala-like taxon in Microtus pennsylvanicus (Ord) (hereafter PMM) and P. kalelai (Tenora, Haukisalmi & Henttonen, 1985) in Clethrionomys rufocanus (Sundevall) using uni- and multivariate (discriminant analysis) morphometric methods. P. kalelai was found to be most divergent among these three taxa using both uni- and multivariate analyses, supporting its status as an independent species. Most of the observed metrical differences between PMG and PMM were found to reflect the larger body size of PMM; the relative size of organs (ratios) did not usually differ significantly between these taxa. The length/width ratio of mature segments was found to be the only size-independent feature that reliably separates PMG and PMM. Since PMM and PMG are also qualitatively very similar, e.g. with respect to the distribution of testes and morphology of the terminal genital ducts, the present analysis could not provide conclusive evidence against their conspecificity. However, the morphological variability and wide host and geographical distribution of P. macrocephala-like cestodes in North American voles of the genus Microtus, and the patterns of host-specificity within Paranoplocephala, suggest that this complex probably includes several species. PMID- 12567011 TI - Polystomatidae (Monogenea) of Southern African Anura: Eupolystoma vanasi n. sp. parasitic in Schismaderma carens (Smith). AB - Eupolystoma vanasi is described as a new species of the Polystomatidae parasitic in the urinary bladder of Schismaderma carens in Northern Province and KwaZulu Natal Province, South Africa. This is the third Eupolystoma species described from Africa and the first polystomatid from Schismaderma, an anuran genus that is primitive with respect to the other African bufonids in which Eupolystoma has been recorded. The species is distinguished by body size (this is the largest Eupolystoma known; mean length of adults 6 mm), by genital spine number (4 in comparison with 6-9 in other species), marginal hooklet length (greater than in other African species), and by the small size of the ovary and testis. In a sample of 27 toads, 37% were infected with up to 130 parasites per host (mean intensity 37). Worm burdens of this magnitude are exceptional amongst polystomatids in general but are characteristic of Eupolystoma, where there may be repeated re-infection of adult hosts and, uniquely, a direct, internal cycle of auto-infection. PMID- 12567012 TI - Woldemar Mobitz. PMID- 12567013 TI - Spike-based synaptic plasticity and the emergence of direction selective simple cells: mathematical analysis. AB - In the companion paper we presented extended simulations showing that the recently observed spike-timing dependent synaptic plasticity can explain the development of simple cell direction selectivity (DS) when simultaneously modifying the synaptic strength and the degree of synaptic depression. Here we estimate the spatial shift of the simple cell receptive field (RF) induced by the long-term synaptic plasticity, and the temporal phase advance caused by the short term synaptic depression in response to drifting grating stimuli. The analytical expressions for this spatial shift and temporal phase advance lead to a qualitative reproduction of the frequency tuning curves of non-directional and directional simple cells. In agreement with in vivo recordings, the acquired DS is strongest for test gratings with a temporal frequency around 1-4 Hz. In our model this best frequency is determined by the width of the learning function and the time course of depression, but not by the temporal frequency of the 'training' stimuli. The analysis further reveals the instability of the initially symmetric RF, and formally explains why direction selectivity develops from a non directional cell in a natural, directionally unbiased stimulation scenario. PMID- 12567014 TI - Clustering in small networks of excitatory neurons with heterogeneous coupling strengths. AB - Excitatory coupling with a slow rise time destabilizes synchrony between coupled neurons. Thus, the fully synchronous state is usually unstable in networks of excitatory neurons. Phase-clustered states, in which neurons are divided into multiple synchronized clusters, have also been found unstable in numerical studies of excitatory networks in the presence of noise. The question arises as to whether synchrony is possible in networks of neurons coupled through slow, excitatory synapses. In this paper, we show that robust, synchronous clustered states can occur in such networks. The effects of non-uniform distributions of coupling strengths are explored. Conditions for the existence and stability of clustered states are derived analytically. The analysis shows that a multi cluster state can be stable in excitatory networks if the overall interactions between neurons in different clusters are stabilizing and strong enough to counter-act the destabilizing interactions between neurons within each cluster. When heterogeneity in the coupling strengths strengthens the stabilizing inter cluster interactions and/or weakens the destabilizing in-cluster interactions, robust clustered states can occur in excitatory networks of all known model neurons. Numerical simulations were carried out to support the analytical results. PMID- 12567015 TI - Propagating waves in visual cortex: a large-scale model of turtle visual cortex. AB - This article describes a large-scale model of turtle visual cortex that simulates the propagating waves of activity seen in real turtle cortex. The cortex model contains 744 multicompartment models of pyramidal cells, stellate cells, and horizontal cells. Input is provided by an array of 201 geniculate neurons modeled as single compartments with spike-generating mechanisms and axons modeled as delay lines. Diffuse retinal flashes or presentation of spots of light to the retina are simulated by activating groups of geniculate neurons. The model is limited in that it does not have a retina to provide realistic input to the geniculate, and the cortex and does not incorporate all of the biophysical details of real cortical neurons. However, the model does reproduce the fundamental features of planar propagating waves. Activation of geniculate neurons produces a wave of activity that originates at the rostrolateral pole of the cortex at the point where a high density of geniculate afferents enter the cortex. Waves propagate across the cortex with velocities of 4 microm/ms to 70 microm/ms and occasionally reflect from the caudolateral border of the cortex. PMID- 12567016 TI - On the integration of subthreshold inputs from Perforant Path and Schaffer Collaterals in hippocampal CA1 pyramidal neurons. AB - Using a realistic model of a CA1 hippocampal pyramidal neuron, we make experimentally testable predictions on the roles of the non-specific cation current, I(h), and the A-type Potassium current, I(A), in modulating the temporal window for the integration of the two main excitatory afferent pathways of a CA1 neuron, the Schaffer Collaterals and the Perforant Path. The model shows that the experimentally observed increase in the dendritic density of I(h) and I(A) could have a major role in constraining the temporal integration window for these inputs, in such a way that a somatic action potential (AP) is elicited only when they are activated with a relative latency consistent with the anatomical arrangement of the hippocampal circuitry. PMID- 12567017 TI - Spike correlation measures that eliminate stimulus effects in response to white noise. AB - When measured in response to non-repeating white noise, standard covariance measures of two neuronal spike trains contain components due simply to a shared stimulus. We argue that, without stimulus repeats, model-free measures cannot in general remove these stimulus-induced components. We present spike correlation measures that eliminate them when the neural response can be approximated by a linear-nonlinear system. One of these measures fully characterizes the correlations in the special case that all remaining correlations are due to small reciprocal connections between the neurons. In addition, we demonstrate that the proposed measures can give accurate results with a more realistic, integrate-and fire model of neural response, provided that it is driven like a linear-nonlinear system. PMID- 12567019 TI - Short overview of potential gene therapy approaches in orthopedic spine surgery. PMID- 12567020 TI - Rehabilitation following first-time lumbar disc surgery: a systematic review within the framework of the cochrane collaboration. AB - STUDY DESIGN: A systematic review of randomized controlled trials. BACKGROUND: Although several rehabilitation programs, physical fitness programs, or protocols regarding instruction for patients to return to work after lumbar disc surgery have been suggested, little is known about the efficacy of these treatments, and there are still persistent fears of causing reinjury, reherniation, or instability. OBJECTIVES: The objective of this systematic review was to evaluate the effectiveness of active treatments that are used in the rehabilitation after first-time lumbar disc surgery. METHODS: The authors searched the MEDLINE, Embase, and Psyclit databases up to April 2000 and the Cochrane Controlled Trials Register 2001, issue 3. Both randomized and nonrandomized controlled trials on any type of active rehabilitation program after first-time disc surgery were included. Two independent reviewers performed the inclusion of studies, and two other reviewers independently performed the methodologic quality assessment. A rating system that consists of four levels of scientific evidence summarizes the results. RESULTS: Thirteen studies were included, six of which were of high quality. There is no strong evidence for the effectiveness for any treatment starting immediately postsurgery, mainly because of the lack of good quality studies. For treatments that start 4 to 6 weeks postsurgery, there is strong evidence (level 1) that intensive exercise programs are more effective on functional status and faster return to work (short-term follow-up) as compared to mild exercise programs, and there is strong evidence (level 1) that on long-term follow-up there is no difference between intensive exercise programs and mild exercise programs with regard to overall improvement. For all other primary outcome measures for the comparison between intensive and mild exercise programs, there is conflicting evidence (level 3) with regard to long-term follow-up. Furthermore, there is no strong evidence for the effectiveness of supervised training as compared to home exercises. There is also no strong evidence for the effectiveness of multidisciplinary rehabilitation as compared to usual care. There is limited evidence (level 3) that treatments in working populations that aim at return to work are more effective than usual care with regard to return to work. Also, there is limited evidence (level 3) that low-tech and high-tech exercises, started more than 12 months postsurgery, are more effective in improving low-back functional status as compared to physical agents, joint manipulations, or no treatment. Finally, there is no strong evidence for the effectiveness of any specific intervention when added to an exercise program, regardless of whether exercise programs start immediately postsurgery or later. None of the investigated treatments seem harmful with regard to reherniation or reoperation. CONCLUSIONS: There is no evidence that patients need to have their activities restricted after first-time lumbar disc surgery. There is strong evidence for intensive exercise programs (at least if started about 4-6 weeks postoperative) and no evidence they increase the reoperation rate. It is unclear what the exact content of postsurgery rehabilitation should be. Moreover, there are no studies that investigated whether active rehabilitation programs should start immediately postsurgery or possibly 4 to 6 weeks later. PMID- 12567018 TI - A cooperation and competition based simple cell receptive field model and study of feed-forward linear and nonlinear contributions to orientation selectivity. AB - We present a model for development of orientation selectivity in layer IV simple cells. Receptive field (RF) development in the model, is determined by diffusive cooperation and resource limited competition guided axonal growth and retraction in geniculocortical pathway. The simulated cortical RFs resemble experimental RFs. The receptive field model is incorporated in a three-layer visual pathway model consisting of retina, LGN and cortex. We have studied the effect of activity dependent synaptic scaling on orientation tuning of cortical cells. The mean value of hwhh (half width at half the height of maximum response) in simulated cortical cells is 58 degrees when we consider only the linear excitatory contribution from LGN. We observe a mean improvement of 22.8 degrees in tuning response due to the non-linear spiking mechanisms that include effects of threshold voltage and synaptic scaling factor. PMID- 12567022 TI - Biomechanical evaluation of cervical double-door laminoplasty using hydroxyapatite spacer. AB - STUDY DESIGN: In vitro three-dimensional kinematic changes after double-door cervical laminoplasty, with and without spacer, and laminectomy were studied in a human cadaveric model. OBJECTIVES: To evaluate the effects of multilevel double door laminoplasty and laminectomy as compared with the intact and to assess the influence of the spinous process spacer on the stability of the cervical spine. SUMMARY OF BACKGROUND DATA: Double-door type cervical laminoplasty has been widely used in the treatment of multisegmental stenotic conditions. However, its biomechanical advantages over laminectomy remain controversial. Also, the biomechanical effects of spacers between the split laminae have not been investigated. METHODS: Using fresh cadaveric C2-T1 specimens, sequential injuries were created in the following order: intact, double-door laminoplasty (C3-C6) with insertion of hydroxyapatite spacers, laminoplasty without spacer, and laminectomy. Motions of each vertebra in each injury status were measured in six loading modes: flexion, extension, right and left lateral bending, and right and left axial rotation.RESULTS Cervical laminectomy showed significant increase in motion compared with intact control in flexion [25% (P < 0.001)], extension [19% (P < 0.05)], and axial rotation [24% (P < 0.001)] at maximum load. Double-door laminoplasty with hydroxyapatite spacer indicated no significant difference in motion in all loading modes compared with intact. Laminoplasty without spacer showed intermediate values between laminoplasty with spacer and laminectomy in all loading modes. Initial slack of each injury status showed trends similar to that of maximum load, although mean percent changes of laminectomy and laminoplasty without spacer were greater than that of maximum load. CONCLUSIONS: Double-door laminoplasty with hydroxyapatite spacer appears to restore the motion of the decompressed segment back to its intact state in all loading modes. The use of HA spacers well contributes to maintaining the total stiffness of cervical spine. In contrast, laminectomy seems to have potential leading postoperative deformity or instability. PMID- 12567021 TI - Overcoming the immune response to permit ex vivo gene therapy for spine fusion with human type 5 adenoviral delivery of the LIM mineralization protein-1 cDNA. AB - STUDY DESIGN: An animal study in immune competent rabbits and athymic rats was conducted. OBJECTIVES: To develop an animal model for simulation of previous human Type 5 adenovirus (Ad5) exposure, to determine the impact of adenoviral pre exposure on spine fusion induced with ex vivo Ad5-LMP-1, and to test strategies for overcoming any potential immune response. SUMMARY OF BACKGROUND DATA: Cells transduced with adenovirus containing the osteoinductive LMP-1 cDNA (Ad5-LMP-1) can induce spine fusion in rabbits. Because up to 80% of the human population has been exposed to adenovirus, immune responses to the vector may limit this strategy in humans. Few studies have modeled previous adenoviral exposure and tested strategies to circumvent it. METHODS: Adult New Zealand white rabbits were injected with 10 or 10 viral particles of Ad5-LacZ. At 4 or 16 weeks after Ad5 injection, autologous buffy coats were prepared from peripheral blood, and 4 million cells per side were infected ex vivo for 10 minutes with Ad5-LMP-1 (multiplicity of infection = 4). Cells were implanted on a collagen matrix instead of an autograft for posterolateral lumbar arthrodesis. Unimmunized rabbits served as control subjects. Additional immunized rabbits underwent arthrodesis at 4 weeks with increased cell number (10 million) and viral dose (multiplicity of infection = 10), or with both parameters increased. The rabbits were killed at 4 weeks, and the spines were assessed by palpation and radiograph. A parallel study was performed in athymic rats using immunized rabbits for the donor cells. RESULTS: All the unimmunized rabbits had solid spine fusions. None of the rabbits arthrodesed 4 weeks after Ad5 pre-exposure achieved fusion. At 4 weeks after Ad5 exposure, increasing the multiplicity of infection to 10 did not overcome the immune response (0/3 fused), but increasing the cell number to 10 million (2/3 fused) or increasing both cell number and multiplicity of infection (3/3 fused) did overcome the immune effects. Delaying arthrodesis until 16 weeks after Ad5 pre-exposure also overcame the immune response (3/3 fused). Similar results were seen in the athymic rat ectopic implant model, suggesting that the immune effect was mediated by humoral antibodies rather than a T-cell response. CONCLUSIONS: Two model systems were developed that simulate previous exposure to human Ad5 and could separate the cellular and humoral components of the response. There was a dose-dependent inhibition of ex vivo Ad5-LMP-1 gene transfer to cells from animals previously exposed to human Ad5. Data suggested that the inhibition of Ad5 infection was caused by humoral antibodies rather than a T-cell-based response. Minor modifications in the gene transfer protocol, such as doubling the viral dose or number of cells infected, or increasing the infection time, could overcome the immune response for an ex vivo approach. PMID- 12567023 TI - The biomechanical effects of spondylolysis and its treatment. AB - STUDY DESIGN: Biomechanical analysis of the level above pars defects was performed using calf lumbar spines. OBJECTIVES: To evaluate whether complete spondylolysis contributes to the pathology of the upper adjacent motion segment to the pars defect. SUMMARY OF BACKGROUND DATA: It is well recognized that patients with spondylolysis show a higher incidence of spondylolisthesis or degenerative disc changes at the level of the pars defects. However, some authors have referred to the fact that disc damage may occur at the level above the defect and give rise to symptoms. However, no previous studies have been directed to the kinematic influence on the upper adjacent segment to pars defects. METHODS: Nine fresh-frozen calf lumbar spines were used for this study. The bony defects were created on the L4 pars articularis bilaterally. Three linear extensometers and one specially designed angular extensometer were mounted across the L3-L4 and L4-L5 motion units. Nondestructive static loads, including axial compression, flexion-extension, and axial rotation, were applied on the specimens in four different conditions as follows: 1) intact spine; 2) bilateral pars defects on the L4 laminae; 3) pars defect repair with Buck technique; and 4) pedicle screw-rod fixation at L4-L5 after removal of the interarticular screws. Testing was performed on a material testing machine (MTS 858 Bionix test system, Minneapolis, MN), and load-displacement curves were recorded with the extensometers. Each test was performed for over five full sinusoidal loading cycles, and data from the fifth cycle were collected and analyzed. RESULTS: After creating the pars interarticularis defects at L4, mobility at both the L3-L4 and L4-L5 motion units were increased in all loading conditions. The normalized range of motion (% ROM) as compared with the intact specimens showed that the pars defects increased the mobility at the upper adjacent level (L3-L4) to 106.4% in flexion-extension and to 120.1% in axial rotation; the differences were significant (P < 0.01). Consequently, the increased mobility was stabilized by applying Buck screws through the defects on both sides; however, the effect was not statistically significant. Furthermore, pedicle screw-rod fixation applied at the L4-L5 segment increased the intervertebral motion at the upper adjacent level, and % ROM in axial rotation was significantly increased to 119.2% of the intact spine (P < 0.05). Comparing the treatments' effects on the L3-L4 segment and that on L4-L5, the Buck screws restored the stability of both segments to the level of the intact spine, whereas the pedicle screw system limited the motion of L4-L5 motion and, on the contrary, increased the L3-L4 motion. CONCLUSIONS: This biomechanical study exhibited that bilateral pars interarticularis defects increased the intervertebral mobility, not only at the involved level but also at the upper adjacent level to the lysis. The increased mobility at the upper segment was reduced by the Buck screw technique. However, this was increased again by the pedicle screw system applied on the involved segment. If clinically applicable, fixation of the pars defect alone appears to cause less adjacent level mechanical stress than pedicle screw-rod motion segment fixation. PMID- 12567024 TI - Anterior occiput to axis screw fixation: part II: a biomechanical comparison with posterior fixation techniques. AB - STUDY DESIGN: This biomechanical study used flexibility testing on fresh-frozen human cadaveric specimens (occiput to C3) and compared the range of motion and neutral zone for three occipitocervical fixation techniques. OBJECTIVES: To contrast the stabilization provided by a new technique of anterior occipitocervical screw fixation with two other commonly used posterior occipitocervical fixation techniques. SUMMARY OF BACKGROUND DATA: There are no published reports describing this novel technique of anterior occipitocervical screw fixation. METHODS: Six human occipitocervical spine specimens were mounted in a custom-designed, spine-testing machine that applied a pure moment in flexion extension, lateral bending, and axial rotation. The specimens were tested intact, after an odontoid osteotomy with capsular injury, and after each of three fixation methods: posterior wiring, posterior plate fixation with C1-C2 transarticular screws, and finally with anterior occipitocervical screws. Intervertebral motion was measured with an optoelectronic measurement system, and the range of motion and neutral zone were the kinematic variables measured and used for analysis. RESULTS: In flexion and extension testing, the posterior plate with transarticular screws provided greater stabilization than posterior wiring or anterior occipitocervical screws. In lateral bending and rotation, the anterior screws were similarly effective to the posterior plate, both of which were more effective than posterior wiring. CONCLUSION: The anterior screw fixation technique was as effective as a posterior plate with transarticular screws in stabilizing between the occiput and C2 in axial rotation and lateral bending. In extension and flexion, the anterior screw technique was not as effective as a posterior plate with transarticular screws in providing stability. PMID- 12567025 TI - Histology of tissues within retrieved human titanium mesh cages. AB - STUDY DESIGN: Histologic documentation of contents within retrieved, failed human titanium mesh cages. OBJECTIVES: The purpose of this study is to describe the contents of a series of retrieved, clinically failed titanium mesh cages from human patients. SUMMARY OF BACKGROUND DATA: The use of interbody fusion cages is gaining rapid acceptance, but there is little histologic documentation of tissue within retrieved cages. METHODS: Eleven Titanium Surgical Mesh Cages (Harms; DePuy AcroMed, Raynham, MA) retrieved from 10 patients were histologically analyzed. Indications for cage retrieval included failed fusion or failed fusion with instrumentation failure. The cages had been in situ from 2 to 47 months. Histologic sections were reviewed qualitatively, and the approximate percent of area in the cage occupied by viable bone, necrotic bone, fibrocartilage, hyaline cartilage, fibrous tissue, and bone graft substitute was visually estimated. Particles of metal debris were estimated by a semiquantitative scoring system. RESULTS: All cages except one showed evidence of vascular ingrowth and areas of histologically viable bone, representing incorporating bone graft. At least a few particles of debris were present in 9 of 11 cages. Fibrocartilage of probable intervertebral disc origin ranged from 0% to 70% of the available area. Several cages contained small seams of fibrocartilage connecting segments of bone in a pattern that suggested a response to motion. CONCLUSIONS: Mean viable bone area within 11 retrieved, human titanium mesh cages was approximately 31%. Seams of fibrocartilage within the cages may represent tissue differentiation in response to bending or compressive load. PMID- 12567027 TI - Direct stabilization of lumbar spondylolysis with a hook screw: mean 11-year follow-up period for 113 patients. AB - STUDY DESIGN: A retrospective study involving 113 patients with a spondylolysis who underwent direct repair with a hook screw was conducted. OBJECTIVES: Spondylolysis with instability can produce low back pain. In adults, fusion of the affected level is the gold standard of treatment. In the adolescent, direct repair of the lysis can save a functional segment. SUMMARY OF BACKGROUND DATA: It has been proved that direct repair of spondylolysis can accomplish restitutio ad integrum for a functional segment. So far, only short-term results are available in the literature. METHODS: This study involved 113 patients who underwent direct repair of spondylolysis with a hook screw according to Morscher. After an average follow-up period of 11 years, the patients were reviewed. The main issue was the question of fusion. For this purpose, plain radiographs were taken. For doubtful cases or for the patient experiencing pain, conventional tomography scans or computed tomography scans were taken. RESULTS: The pseudarthrosis rate was 13.3%. The fusion rate for patients younger than 14 years was higher than that for older patients, especially those older than 20 years. CONCLUSIONS: Direct repair of spondylolysis can be recommended for patients with a growing skeleton, a slight slip, an instability, or failure of conservative treatment. Thereby, a functional segment can be saved with a relatively small operation. PMID- 12567028 TI - Neonatal kyphectomy in the patient with myelomeningocele. AB - STUDY DESIGN: A retrospective cohort study was used to investigate a group of neonates with myelomeningocele who had a kyphectomy performed in conjunction with dural sac closure during the first few days of life. OBJECTIVES: To assess the effectiveness of operative intervention in the neonatal period to correct the kyphotic deformity in the patient with myelomeningocele and to monitor its long term results. SUMMARY OF BACKGROUND DATA: Orthopedic management originally focused on the immediate treatment of the kyphotic deformity in the infant with myelomeningocele. However, there has been a movement toward postponing surgical treatment of the kyphos until a later age. This study included the longest follow up of the largest group of neonates that a single surgeon has managed surgically since the treatment of this condition was originally described. METHODS: The radiographic and clinical results for all neonates treated with a kyphectomy at the time of myelomeningocele closure between 1980 and 2000 were analyzed. RESULTS: Neonatal kyphectomy was performed on nine males and two females. The average preoperative kyphotic angle measured 67 degrees. The average initial correction was 77 degrees, and the average loss of correction at follow-up assessment was 55 degrees. There were no serious complications, and wound closure was successful in all patients. One patient required a repeat kyphectomy and posterior spinal fusion at the age of 9 years and 2 months. The average follow-up period was 7 years and 4 months (range 44-174 months). CONCLUSIONS: Kyphectomy performed at the time of dural sac closure in the neonate is a safe procedure with excellent initial correction. Eventual recurrence is expected despite the procedure. However, it occurs in the form of a longer, more rounded deformity that is less technically demanding. PMID- 12567029 TI - The crankshaft phenomenon after posterior spinal arthrodesis for congenital scoliosis: a review of 54 patients. AB - STUDY DESIGN: Retrospective chart and radiographic reviews were conducted. OBJECTIVE: To identify the incidence of and any possible risk factors for the crankshaft phenomenon after posterior spinal arthrodesis for congenital scoliosis. SUMMARY OF BACKGROUND DATA: Studies have shown the crankshaft problem to be common after posterior arthrodesis for infantile and juvenile idiopathic scoliosis, but the few reports available show it to be much less common for congenital scoliosis. METHODS: This study chose children fused before the pubertal growth spurt, all classified as Risser 0 and with open triradiate cartilages. These children were followed to the end of their growth (mean follow up period 12 years). Several measurement parameters were used for evaluation. RESULTS: The crankshaft problem, measured as a Cobb angle increase of more than 10 degrees, was seen in 15% of the 54 patients. There was a positive correlation with earlier surgery and larger (>50 degrees) curves. No other positive correlations could be identified. CONCLUSIONS: Crankshafting was observed in 15% of the patients, more often with larger curves and earlier fusions. PMID- 12567030 TI - Radiologic evaluation of adjacent superior segment facet joint violation following transpedicular instrumentation of the lumbar spine. AB - STUDY DESIGN: The location of pedicle screws in relation to adjacent superior segment facet joints in 106 patients after lumbar spinal fusion was assessed using computed tomography and plain radiographs. OBJECTIVES: To document the incidence of adjacent superior segment facet joint violation following transpedicular instrumentation in the lumbar spine. SUMMARY OF BACKGROUND DATA: Review of the literature failed to show any documented study examining this incidence. METHODS: A total of 106 patients (212 top-level facet screws) undergoing lumbar spinal fusion surgery using transpedicular instrumentation between 1996 and 1999 were prospectively evaluated with a computed tomography scan and plain radiographs at 6 months following surgery. These were blindly and independently evaluated by a consultant radiologist and a spinal research fellow to document the incidence of superior segment facet joint violation. Calculation of the kappa coefficient and chi2 analysis were carried out. RESULTS: The spinal research fellow noted the incidence of facet joint violation on the computed tomography scan to be present in 20% of the screws and 32% of the patients, whereas the consultant radiologist noted this to be the case in 23% and 35%, respectively. The kappa coefficient for computed tomography scan was 0.88, whereas for the plain radiographs it was 0.39. The incidence of facet joint violation was noted to be independent of the sex, level, and diagnosis. There was also an almost uniform incidence in each of the years from 1996 to 1999. CONCLUSION: Facet joint violation occurred in just >30% of the patients and 20% of the screws in this study. This, therefore, raises the theoretical possibility of long-term deterioration in the clinical results following the use of transpedicular instrumentation. PMID- 12567031 TI - The motor-evoked potentials elicited from the deltoid muscle by transcranial magnetic stimulation with a standardized facilitation: the potential diagnostic utility for C5 radiculopathy. AB - STUDY DESIGN: We investigated the effects of standardized facilitation on stability in amplitude of the motor-evoked potentials from the deltoid muscle after magnetic stimulation over the scalp in normal study participants. OBJECTIVES: To test the potential of this technique in evaluating deltoid weakness and recovery after C5 radiculopathy. SUMMARY OF BACKGROUND DATA: Most previous motor-evoked potential studies have emphasized the value of the latency rather than amplitude, probably because the amplitude varies much more than the latency, even in the same individual. METHODS: Twenty right-handed healthy men underwent deltoid motor-evoked potential studies on three successive occasions at intervals of >2 weeks. A digital readout of the shoulder abduction force allowed the study participant to adjust voluntary contraction to 20% of the maximal force at the moment of stimulation. The deltoid motor-evoked potentials were compared with the M responses recorded from the relaxed deltoid after electrical stimulation of the brachial plexus. RESULTS: Despite considerable individual variability in the amplitude of deltoid motor-evoked potentials, a standardized facilitation yielded a relatively small variation of amplitude between sessions in the same individual, showing an intersession variability index of 9.1 +/- 7.9%. The side-to-side amplitude ratio, averaging 82 +/- 12%, also provides a more reliable measure in evaluating unilateral deltoid weakness than the absolute value. The measures on the left showed significantly higher amplitude than on the right for the deltoid motor-evoked potentials but not for the M responses. CONCLUSIONS: The changes of the motor-evoked potential amplitude exceeding 25% between sessions and side-to-side amplitude ratios <60% can be outside the normal limits. The asymmetry of deltoid motor-evoked potentials in favor of the nondominant side is of physiologic interest. PMID- 12567032 TI - Derecruitment of the lumbar musculature with fatiguing trunk extension exercise. AB - STUDY DESIGN: This was a descriptive study involving 20 healthy individuals. OBJECTIVES: To evaluate the neuromuscular activation patterns of the lumbar paraspinal and hip extensor muscles during isotonic trunk extension exercise. SUMMARY OF BACKGROUND DATA: Few studies have evaluated the effect of muscle fatigue on the lumbar musculature during isotonic exercise. METHODS: Electromyographic activity was recorded continuously from the lumbar paraspinal, gluteus maximus, and biceps femoris muscles during isotonic trunk extension exercise performed to muscular failure. Root mean squared electromyography was determined over the concentric portion of each repetition, and polynomial regression analysis was used to describe the association between fatigue and the recruitment patterns. RESULTS: The lumbar paraspinals demonstrated an increase in the electromyogram signal up to 57.9% of maximal fatigue, at which point decrements in electromyography were observed (lumbar [quadratic curve] R2 = 0.0807, SEE = 0.228; beta2 = -8.245(-5)) (P < 0.000). Associated with fatigue, the gluteus maximus demonstrated an increase in electromyography, with an exponential breakpoint occurring at 35.9% of maximal fatigue (gluteus maximus [quadratic curve]: R2 = 0.5059, SEE = 0.865; beta2 = 0.00017) (P = 0.014). The biceps femoris demonstrated a linear increase in electromyography with fatigue (R2 = 0.4667, SEE = 0.284; beta2 = 0.0091) (P < 0.000). To further investigate the derecruitment of the lumbar extensors associated with fatigue, study participants were analyzed individually with regression analyses. Results revealed that the majority of study participants (68.5%) demonstrated a significant decrease (quadratic bend) in lumbar electromyography, with decrements in muscle activity beginning at 53% of maximum. CONCLUSION: During fatiguing trunk extension exercise, an increase in the lumbar paraspinal electromyogram signal occurs up to approximately 55% of maximum fatigue, at which point a decrease in electromyography is observed. Associated with this derecruitment is a concomitant increase in hip extensor muscle activity, suggesting that as the lumbar musculature becomes fatigued, these muscles allow for continuation of the exercise. PMID- 12567033 TI - Utility of intraoperative ultrasound for tumors of the cauda equina. AB - STUDY DESIGN: A retrospective chart review of patients with tumors of the cauda equina was performed. OBJECTIVES: To assess the role of intraoperative ultrasound for detecting tumor migration and modifying the approach to cauda equina tumors. SUMMARY OF BACKGROUND DATA: Rostral migration of cauda equina tumors relative to preoperative imaging is well described. The utility of intraoperative ultrasound to identify tumor migration and to modify the surgical approach was analyzed. METHODS: The medical records of nine consecutive patients undergoing biopsy or resection of a cauda equina tumor diagnosed by magnetic resonance imaging at the authors' institution from January 1994 through December 2000 were reviewed. Four patients with tumors of the cauda equina underwent transdural sonography of the intraspinal compartment after initial laminectomy and before durotomy. RESULTS: In three patients, the findings on intraoperative ultrasound showed rostral migration of the lesion with respect to the preoperative magnetic resonance imaging, necessitating additional laminectomy and modifying the site of durotomy. One tumor had migrated rostrally one full bony vertebral segment, whereas two tumors each had migrated one half of a bony vertebral segment. Pathology examination in the three patients whose lesions had migrated rostrally demonstrated a schwannoma in every case. In one patient, intraoperative ultrasound showed no lesion migration with respect to the preoperative magnetic resonance imaging, and no modifications to the surgical approach were needed. Pathology examination of this patient demonstrated an ependymoma. CONCLUSIONS: Intraoperative ultrasound was useful in documenting rostral migration of cauda equina tumors, and facilitated appropriate laminectomy and durotomy. The technique may be particularly valuable for schwannomas of the cauda equina, which seem to have a greater potential for rostral migration. PMID- 12567035 TI - Patterns and perceptions of care for treatment of back and neck pain: results of a national survey. AB - STUDY DESIGN: We conducted a nationally representative random household telephone survey to assess therapies used to treat back or neck pain. OBJECTIVES: The main outcome was complementary therapies used in the last year to treat back or neck pain. SUMMARY OF BACKGROUND DATA: Back pain and neck pain are common medical conditions that cause substantial morbidity. Despite the presumed importance of complementary therapies for these conditions, studies of care for back and neck pain have not gathered information about the use of complementary therapies. METHODS: Our nationally representative survey sampled 2055 adults. The survey gathered detailed information about medical conditions, conventional and complementary therapies used to treat those conditions, and the perceived helpfulness of those therapies. RESULTS: We found that of those reporting back or neck pain in the last 12 months, 37% had seen a conventional provider and 54% had used complementary therapies to treat their condition. Chiropractic, massage, and relaxation techniques were the most commonly used complementary treatments for back or neck pain (20%, 14%, and 12%, respectively, of those with back or neck pain). Chiropractic, massage, and relaxation techniques were rated as "very helpful" for back or neck pain among users (61%, 65%, and 43%, respectively), whereas conventional providers were rated as "very helpful" by 27% of users. We estimate that nearly one-third of all complementary provider visits in 1997 (203 million of 629 million) were made specifically for the treatment of back or neck pain. CONCLUSIONS: Chiropractic, massage, relaxation techniques, and other complementary methods all play an important role in the care of patients with back or neck pain. Treatment for back and neck pain was responsible for a large proportion of all complementary provider visits made in 1997. The frequent use and perceived helpfulness of commonly used complementary methods for these conditions warrant further investigation. PMID- 12567037 TI - The effectiveness of standard care, early intervention, and occupational management in worker's compensation claims. AB - STUDY DESIGN: A retrospective and prospective cohort. OBJECTIVES: To compare the effectiveness of occupational intervention, early intervention, and standard care in the management of Worker's Compensation injury claims. SUMMARY OF BACKGROUND DATA: The current management of occupational back pain and work-related upper extremity disorders with either standard care or early intervention appears to be ineffective. METHODS: A retrospective cohort compared injury claim incidence, duration, and costs between one company with access to standard care and another similar company with access to early intervention. A prospective cohort looked at the effect of one company changing from standard care to occupational management in comparison with the control group with early intervention. Survival analysis was used to attempt to explain differences in injury claim duration. RESULTS: Standard care resulted in lower injury claim incidence, duration, and costs than early intervention, whereas occupational management resulted in lower injury claim incidence, duration, and costs than standard care. The covariates of physical therapist involvement, chiropractor involvement, injury severity, and relationship between Worker's Compensation and the employer were associated with delayed time to claim closure in the company with access to early intervention with the most important covariate being physical therapist involvement (hazard rate ratio 19.88, 95% confidence interval 7.95-39.77). Only the covariate of injury severity was associated with delayed time to claim closure in the company with access to occupational management (hazard rate ratio 1.67, 95% confidence interval 1.05-27.20). CONCLUSIONS: It is recommended that an occupational management approach, in comparison with standard care or early intervention, be considered for management of occupational injuries. PMID- 12567038 TI - A new halo-pelvic apparatus. AB - STUDY DESIGN: A technical report of a new, versatile halo-pelvic apparatus for use in spinal immobilization is presented. OBJECTIVES: To show how simple modifications of existing external fixation devices allow a halo-pelvic fixation apparatus to be constructed. SUMMARY OF BACKGROUND DATA: Halo-pelvic fixation has been used successfully in the past for immobilization and correction of severe spinal deformities such as tuberculous kyphosis. With the advent of new spinal instrumentation systems and techniques, external immobilization of the whole spine is rarely indicated. However, on rare occasions, spine surgeons may be presented with cases in which no internal fixation has been possible. METHODS: Halo-skull fixation and pelvis fixation were performed using standard techniques. A simple custom-made halo connection bar was used to connect the halo to uprights from the pelvis external fixator. Because the majority of the system makes use of standard external fixation equipment, it is easy to apply and adjust. RESULTS: Between 1997 and 2001, this frame was successfully applied to stabilize the spines of three patients with severe tuberculous kyphosis exceeding 100 degrees. CONCLUSIONS: One simple modification of existing external fixation equipment has allowed a new halo-pelvic device to be constructed and used effectively to immobilize the spine. PMID- 12567039 TI - Americans with Disabilities Act: physician-shareholder practice groups and ADA compliance. AB - This article examines the application of Americans with Disabilities Act requirements to professional associations like physician practice groups. In general, employers with 15 or more full-time employees must comply with the Act. However, the definition of an employee is sometimes unclear, especially as applied to business entities commonly used by physician practice groups. A recent case decided by the United States Court of Appeals for the Ninth Circuit held that physician-shareholders of a professional corporation are employees for Americans with Disabilities Act coverage purposes. Analogous cases in other federal circuits have held differently, likening the "owners" of professional corporations to partners in a partnership, who are not considered employees. Similar questions arise for popular business entities, such as Limited Liability Companies and Limited Liability Partnerships. This article discusses the nature of the business forms commonly used by physician practice groups and how their characteristics impact employee status for Americans with Disabilities Act coverage. It then suggests that examination is useful beyond business formation characteristics to the purpose of the Americans with Disabilities Act and other employment antidiscrimination statutes. PMID- 12567040 TI - Lymphangiomatosis of the spine: two cases requiring surgical intervention. AB - STUDY DESIGN: Two cases of lymphangiomatosis of the spine are presented. OBJECTIVES: To report two cases of lymphangiomatosis of the spine requiring surgical intervention and to review the literature. SUMMARY OF BACKGROUND INFORMATION: Lymphangiomatosis is a rare childhood disease characterized by abnormal lymph tissue at multiple sites. Skeletal and visceral involvement are both common. Prognosis depends on the extent of extraskeletal disease. METHODS: Two cases of lymphangiomatosis causing neural compression and instability at the cervicothoracic junction are presented. Both patients underwent surgical decompression and stabilization. RESULTS: One patient died, whereas the other regained full function and activity. CONCLUSIONS: Surgery is indicated when lymphangiomatosis causes neural compression and instability of the spine. Surgical outcome is strongly influenced by extraskeletal involvement. PMID- 12567041 TI - Acute traumatic lumbosacral dislocation treated by open reduction internal fixation and fusion. AB - STUDY DESIGN: Case report of a patient with acute traumatic lumbosacral dislocation. OBJECTIVE: To report a case of traumatic lumbosacral dislocation treated with open reduction internal fixation and fusion. SUMMARY OF BACKGROUND DATA: To our knowledge, there are only 49 cases reported in the literature of this exceptional lesion. Complete lumbosacral dislocation is a three-column lesion, and therefore, open reduction internal fixation and fusion is recommended. METHOD: We report the case of a 42-year-old man who had a vehicle accident. In addition to other fractures, he suffered an anterior lumbosacral dislocation. The displacement of L5 on S1 was 35%. The patient was surgically treated with open posterior reduction fixation and fusion with good result. RESULTS: Complete fusion was achieved, and at 5 years follow-up, the patient was asymptomatic, and no further slippage has been observed. CONCLUSIONS: A rare case of acute anterior lumbosacral dislocation treated surgically is reported. We consider the surgical treatment for reduction, decompression, stabilization, and fusion as the method of choice in acute cases of this exceptional condition. PMID- 12567042 TI - Anterior occiput-to-axis screw fixation: part I: a case report, description of a new technique, and anatomical feasibility analysis. AB - STUDY DESIGN: A case report of anterior screw fixation from the axis to the occiput is described, as is the surgical technique. The pertinent anatomy is described with a radiographic assessment of the feasibility, safety, and general applicability of this technique. OBJECTIVES: To describe a novel technique of anterior occipitocervical fixation and the pertinent anatomy. SUMMARY OF BACKGROUND DATA: In unique clinical situations where posterior fixation techniques may not be possible or may have already failed, an anterior screw fixation technique may add stability to further attempts at obtaining a posterior arthrodesis. METHODS: A case report is presented, followed by a detailed description of the surgical technique. Ten normal cervical spines had radiographs and computed tomography scans with reformats reviewed to determine screw entry points, target points, and proposed screw trajectories. Following screw insertion in eight fresh frozen human cadaver spine specimens, dissection verified screw location relative to structures at risk. RESULTS: The ideal entry point is located caudal to the C2 superior facet joint in line with the medial third of the C2 superior facet. The screw is directed 25 degrees posteriorly in the sagittal plane and 15 degrees laterally in the coronal plane. The screw tip is located in the posterolateral third of the occipital condyle. Anatomic variation is considerable and makes this technique inadvisable in up to 20% of cases. Structures at risk include the vertebral artery and the hypoglossal nerve. CONCLUSIONS: This new technique of anterior fixation of the atlas to the occiput is feasible and safe if meticulous surgical planning is performed. PMID- 12567043 TI - Acute neurologic deficit associated with a posterior cervical drain: a case report. AB - STUDY DESIGN: Descriptive. OBJECTIVES: To document a rare complication involving the use of a wound drain after cervical laminectomy. SUMMARY OF THE BACKGROUND DATA: No previous reports describe spinal cord compression by a surgical drain resulting in a neurologic deficit. Most texts recommend the use of a drain following this procedure. METHODS: An 80-year-old female underwent cervical laminectomy for myelopathy with initial improvement in symptoms. Approximately 6 hours following surgery, she developed acute onset of quadriparesis when repositioning in bed. Magnetic resonance imaging revealed cord compression by the surgical drain, which was immediately removed. RESULTS: The patient experienced partial improvement of the neurologic deficit. At 18-month follow-up, left handed weakness, left leg spasticity, and neurogenic bladder persist. CONCLUSION: The development of neurologic deficits due to compression by a surgical drain can occur. This complication might be avoided by approximating the neck musculature before placement of the drain and closure of the fascia. PMID- 12567044 TI - Posterior epidural migration of lumbar disc fragments: report of three cases. AB - STUDY DESIGN: Report of three cases. OBJECTIVES: To describe a rare location of intervertebral disc migration. SUMMARY OF BACKGROUND DATA: Migration of sequestered disc fragments to the posterior extradural space is rare, and posterior migration of the free fragments causing cauda equina syndrome is exceptionally rare. METHODS: Three patients with posteriorly migrated epidural disc fragments were evaluated with radiograph, computed tomography, and magnetic resonance imaging and underwent surgery. RESULTS: All of the patients responded well to operative therapy with complete relief of symptoms. CONCLUSIONS: Early surgery should be the first choice of therapy in patients with large posteriorly migrated sequestered disc fragments, to prevent severe neurologic deficits such as cauda equina and conus medullaris syndromes. PMID- 12567045 TI - Heterotopic replantation in mutilating hand injury. AB - To achieve optimal hand function in the reconstruction of a mutilated hand is quite a challenge, and every salvage procedure should be tried, especially for amputated parts that cannot be replanted to their original anatomic sites. Five patients who sustained work-related mutilating hand injuries and who underwent digital heterotopic replantation are reported. Two patients had irreparably crushed, amputated thumbs. One patient (patient 1) received heterotopic replantation of the ring finger to the thumb's position; the other patient (patient 2) received heterotopic replantation of the middle finger to the thumb. Three additional patients received digital heterotopic replantation to enhance hand function. Patient 3 underwent heterotopic replantation of the middle finger to the index finger's position. Patient 4 underwent transplantation of the middle finger to the index finger's position, the ring finger to the middle finger's position, and the little finger to the ring finger's position. In patient 5, the ring finger was transplanted to the index finger's position. In all patients, satisfactory results were achieved in hand function and wound healing. In these patients, the restoration of a functional thumb was the first priority, followed by establishing at least two opposable fingers at the sites where metacarpophalangeal joints were functional. PMID- 12567046 TI - Deep inferior epigastric perforator flap: a modification that simplifies elevation. AB - The deep inferior epigastric perforator flap is accepted worldwide as a reliable and relatively safe technique for autologous breast reconstruction. Preserving the continuity of the rectus muscle, it reduces donor site morbidity, allowing less postoperative pain, a faster recovery, and a reduced hospital stay. Nevertheless, the more complex nature of this type of surgery leads to increased operating time and a demand for more tedious microsurgical dissection. The authors present a modification of the deep inferior epigastric perforator flap harvesting technique that allows a more safe, simple, and faster elevation of the flap. They have used it successfully for breast reconstruction in 3 patients. PMID- 12567047 TI - Free perforator crossover TRAM flap for breast reconstruction. AB - Breast reconstruction using a transverse rectus abdominis musculocutaneous (TRAM) flap has become the preferred method of autogenous reconstruction for most surgeons. The vascular basis of both the superior and inferior vascular pedicles of this flap has been well documented. When a pedicled TRAM flap is based superiorly, the perfusion across the midline to zone 4 and sometimes zone 3 is, at best, variable. Augmentation of the blood supply of the contralateral side with various methods has been reported. The methods include the delay procedure, bipedicled flaps, supercharging, and turbo-charging. The deep inferior epigastric artery is the dominant blood supply, and a microsurgical free TRAM flap based inferiorly provides reliable perfusion, even to zone 4, which obviates the need for many of these maneuvers. It has also been demonstrated that the circulation across the midline in a TRAM flap is primarily by means of a subdermal plexus and that with a previous vertical midline abdominal scar there is virtually no midline crossover at any anastomotic level. Therefore, even with a free TRAM flap based on the dominant inferior pedicle, perfusion across a vertical midline scar is unreliable. As a result, many patients with a vertical midline scar have been denied the best autogenous reconstructive option. The authors present their experience with a free perforator crossover TRAM flap using a constant premuscular branch of the deep inferior epigastric artery and vein that provides many patients who have a previous midline scar with a genuine option for autogenous tissue breast reconstruction. PMID- 12567048 TI - Use of internal mammary vessel perforator as a recipient vessel for free TRAM breast reconstruction. AB - Breast reconstruction is a cosmetically critical procedure for women and it must be undertaken to balance the shape, size, and position of the breast with the other breast. Since the first introduction of the free abdominoplasty flap in 1979, the transverse rectus abdominis musculocutaneous (TRAM) flap technique has been a widely accepted method of breast reconstruction after mastectomy. In breast reconstruction with a free flap, the selection of suitable recipient vessels is the critical decision to be made by the surgeon. The most common recipient vessel for free flap breast reconstruction is the axillary system. However, when used as a recipient, the axillary system may limit flap movement and flexibility in breast shaping. The use of the internal mammary vessels as a recipient site attains ideal breast symmetry. However, the technique requires partial rib resection and eliminates the opportunity for a potential coronary artery bypass graft, which requires the internal mammary artery. Based on these considerations, the selection of suitable recipient vessels constitutes an important requirement for successful free tissue transfer. The authors have performed breast reconstruction with the TRAM flap anastomosed to the internal mammary perforator vessel and conclude that these perforators could be useful as recipient vessels, especially in the case of immediate breast reconstruction with the free TRAM flap. PMID- 12567049 TI - Influence of vascular delay on abdominal wall complications in unipedicled TRAM flap breast reconstruction. AB - Surgically delaying a unipedicled lower abdominal transverse rectus abdominis musculocutaneous (TRAM) flap has been shown to improve flow within the flap. This delay, however, also affects blood supply and drainage of the entire anterior abdominal wall. The purpose of this study was to compare the abdominal complications between surgically delayed and nondelayed TRAM flaps. A retrospective case review of lower abdominal TRAM flap breast reconstructions was performed. A total of 35 patients were included in the study, of whom 15 had undergone delay and 20 had not. The patients were found to be matched by age and body mass index. There was a higher incidence of smokers (past or present) in the delayed series. Despite this, no abdominal flap complications were experienced in those who underwent delay. In the nondelayed series, however, three patients (15%) experienced delayed healing, and two flaps (10%) underwent severe necrosis (p = 0.047, chi-squared test). In the delayed series, one patient (7%) was found to have an abdominal bulge. No hernias were encountered. In contrast, two bulges (10%) and four hernias (20%) were identified in patients in the nonndelayed series (p = 0.6 and 0.09, respectively, chi-squared test). These data suggest that a preliminary delay procedure leads to a reduction in the incidence of abdominal wall complications in unipedicled lower abdominal TRAM flaps. PMID- 12567050 TI - Honey-medicated dressing: transformation of an ancient remedy into modern therapy. AB - Honey has been applied for medicinal purposes since ancient times. Its antibacterial effects have been established during the past few decades. Still, modern medical practitioners hesitate to apply honey for local treatment of wounds. This may be explained by the expected messiness of such local application. Moreover, secondary infectious disease may be caused by contamination of honey with microorganisms. Hence, if honey is to be applied for medicinal purposes, it has to meet certain criteria. The authors evaluated the use and safety of a honey-medicated dressing that was developed to meet these criteria in a feasibility (phase II) study featuring 60 patients with chronic (n = 21), complicated surgical (n = 23), or acute traumatic (n = 16) wounds. In all but 1 patient, it was found easy to apply, helpful in cleaning the wounds, and without side effects. Based on these results, the authors advise to subject this dressing to a randomized, double blind, phase III study. PMID- 12567051 TI - One-stage reconstruction of soft tissue and Achilles tendon defects using a composite free anterolateral thigh flap with vascularized fascia lata: clinical experience and functional assessment. AB - The combined loss of the Achilles tendon with overlying soft tissue is a reconstructive challenge. To achieve acceptable rehabilitation, such patients need skin coverage including functional repair of the Achilles tendon. This article presents four such patients who were treated successfully by means of an anterolateral thigh (ALT) composite flap with vascularized fascia lata. The size of the ALT flaps ranged from 10 to 16 cm in length and 6 to 9 cm in width. All flaps included vascularized fascia lata, which was rolled to serve as vascularized tendon graft (range 8 x 6 cm to 10 x 8 cm) for reconstruction of the Achilles tendon defect. Flap success rate was 100%. All patients could walk and climb stairs without support; however, mild difficulty when running was reported. Functional outcome of the recipient ankle and donor thigh morbidity were investigated by using a kinetic dynamometer comparing reconstructed sides with the healthy contralateral limbs. This assessment was performed in two patients at 2 years postoperatively. In the reconstructed ankles, isokinetic concentric measurements of dorsiflexion and plantar flexion showed a deficit of 30% and 40%, respectively. Functional evaluation of quadriceps femoris muscle contraction forces after free ALT composite flap harvest showed a 10% to 25% deficit. However, there were no difficulties in daily ambulating. In summary, the free composite ALT flap with vascularized fascia lata provides an alternative option for Achilles tendon reconstruction in complex defects. PMID- 12567052 TI - Lateral septoaponeurotic artery: source of bleeding in blepharoplasty performed in asians. AB - In standard blepharoplasty, the orbital septum is opened commonly to enter the preaponeurotic space. This is to anchor the leading edge of the levator palpebral fascia to the skin of a proposed supratarsal fold and to remove supraorbital fat. Incision of the orbital septum is involved occasionally with accidental injury to an inconstant arterial branch at its lateral side, which causes severe bleeding. Anatomic knowledge of the vessel on the orbital septum is conducive to preventing accidental arterial injury. The authors encountered a relatively large artery running vertically at the lateral side of the orbital septum in 50 eyelids of 230 blepharoplasty patients (460 eyelids). They investigated the branch by injecting red latex into the ophthalmic arteries in 20 hemidissected faces of fresh cadavers. At the lateral aspect of the orbital septum, a branch of the superior lateral palpebral artery, which we named the lateral septoaponeurotic artery (LASA), was confirmed in five of 13 specimens. The LASA originates from the superior lateral palpebral artery just after it divides into peripheral and marginal arcades. The LASA pierces the levator aponeurosis and orbital septum at the upper level of the tarsal plate. It anastomoses with a branch of the supraorbital artery at the superior aspect of the orbit. The authors think severe bleeding occurs when the LASA is injured during blepharoplasties. Cautious treatment of the lateral septoaponeurotic vessel minimizes undesirable complications. PMID- 12567053 TI - Short-term postoperative flow changes after free radial forearm flap transfer: possible cause of vascular occlusion. AB - The risk for free flap thrombosis is greatly influenced by blood flow. Postoperative hemodynamic changes in vascular pedicles of the microvascular skin flap have not been reported, however. This study focuses on the intraoperative and postoperative changes in the flow volume in the vascular pedicles of the free forearm flap examined by color Doppler ultrasonography. The arterial flow volume increased continuously until day 7, compared with which, the volume after flap elevation was 36%. On day 1, it reached only 52%. In the venous pedicle, the flow volume through the cutaneous vein was only 37% compared with that through the radial vena comitans after flap elevation, whereas the volume through both veins was equal on day 7. Drastic changes in the flow explain the possible vascular occlusion during the early postoperative period in the free forearm flap transfer. PMID- 12567054 TI - Bimodal distribution of the blood supply to lower abdominal fat: histological study of the microcirculation of the lower abdominal wall. AB - Fat necrosis is a common postoperative complication in transverse rectus abdominis muscle flap breast reconstruction. A histological quantification of the blood supply to the lower abdominal fat was undertaken to define this problem further. One hundred eighty sections, 1.5 cm(2) in size, were studied from one half of the lower abdomen in 10 fresh cadavers. These represent three different depths of fat (superficial, middle, deep) and six different anatomic areas (upper, middle, and lower sectors of the medial and lateral parts of the hemiabdomen). The average diameter, number of blood vessels, and the total vascular area were measured in each section. The average diameter of the blood vessels was greater in the superficial compared with the mid fat layer (p = 0.02). The total vascular area was greater in the deep compared with the mid fat layer (p = 0.01) and the superficial fat layer (p = 0.05). The number of blood vessels was also significantly higher in the deep fat layer compared with the mid fat layer (p = 0.001) and the superficial fat layer (p = 0.01). The lower medial area of the lower anterior abdominal wall contained the largest number and average diameter of blood vessels whereas the mid lateral area contained the smallest number and average diameter; however, this did not reach significance (p = 0.1 and 0.2). The results of this study are in keeping with the bimodal vascular supply to the subcutaneous fat of the anterior abdominal wall. The deep and the superficial fat layers are supplied more richly with vessels with a larger diameter whereas the intermediate fat layer is supplied from the terminal branches of these vessels. PMID- 12567055 TI - Effects of the abdominal midline incision on the survival of the transverse rectus abdominis musculocutaneous flap in rat model. AB - The transverse rectus abdominis musculocutaneous (TRAM) flap, as the standard for breast reconstruction, can be performed in selected patients with operation scars in the abdomen. It is questioned why the unilateral TRAM flaps raised from the abdomen with scars seem to have more blood supply to the periphery. The effect of the abdominal midline incision on the survival of the TRAM flap was studied with the rat model. The survival of the inferiorly based unilateral TRAM flap was compared between a control group and three experimental groups in which initial abdominal midline incision was made 1, 2, or 4 weeks before TRAM flap elevation. The control group had a mean survival of 47.05 +/- 4.52% of their original flap surface area. The viabilities of the other groups were 72.19 +/- 11.05, 76.96 +/- 7.79, 90.26 +/- 3.67%, respectively. Results from all three groups were significantly higher than that of the control group. In the microangiogram of the TRAM flap, the main pedicle (deep epigastric system) and muscle perforators remained unchanged, but the subdermal plexus of the abdominal wall showed increase of size and density after incision. Histological examination also revealed corresponding findings. A midline incision in the abdomen, which was made at 1 or more weeks before unilateral TRAM flap formation, was found to improve the survival of the flap. These data may be explained by increase in the size and density of the subdermal plexus, which delivers the blood flow from the muscle perforators to the expanded area of abdominal wall. Therefore, the process of making the abdominal midline incision can be regarded as a minimal form of a prefabrication procedure, which is one of the well-known delay procedures. PMID- 12567056 TI - Growth difference of multilayer onlay cartilage grafts in rabbit ears. AB - Successful survival and growth of autogenous cartilage graft has been well proved in the experimental studies of Dupertuis and Peer. However, it is still unknown whether multilayer cartilage graft has the same survival and growth as a single layer graft. The authors studied any difference of survival and growth rate between single and multilayer cartilage grafts in rabbits. Single, double, and triple layers of autogenous cartilage grafts were inserted into three separate pockets in a single rabbit ear. The three grafts were left in place for 6 months. They performed this procedures in 10 rabbit ears. Proliferating activity of chondrocytes was measured with a method of proliferating cell nuclear antigen immunolabeling. The authors found that there were more proliferating cell nuclear antigen-positive cartilage cells in the graft than in the host bed, but no difference in weight and thickness of the graft among the three groups. PMID- 12567057 TI - Use of oncogene expression as an independent prognostic marker for primary melanoma. AB - For 25 years, the Breslow depth of primary tumors has remained the most accurate prognostic test of survival for primary melanoma. However a number of studies have indicated that outcome for intermediate-thickness melanomas (0.75-2.49 mm) is often at variance with that predicted by the Breslow depth. This study investigated c-myc oncogene expression in 92 primary tumors of intermediate thickness using flow cytometry. Oncoprotein expression was detected in 87 tumors (95%) with a median positivity of 48% (range, 0%-96%). Survival analysis performed using the Kaplan-Meier method revealed a significant association between oncoprotein positivity and clinical outcome (p < 0.01, log-rank test). Multifactorial analysis of survival using Cox's proportional hazards model revealed c-myc oncoprotein to be an independent prognostic marker more accurate than all other clinicopathological parameters including the Breslow depth (chi(2) = 9.68, p< 0.01). Estimation of c-myc oncoprotein is therefore recommended as a powerful prognostic marker for intermediate-thickness primary melanoma. PMID- 12567058 TI - Early rehabilitation of facial defects using interim removable prostheses: report of two clinical cases. AB - Postsurgical facial defects often pose a challenge to patient rehabilitation. Such defects can have a severe adverse effect on patient perceptions of body image and self-esteem. When immediate surgical repair of the defect is not feasible, an interim removable facial prosthesis may be considered. This prosthesis can be fabricated and placed as soon as several days after surgery to provide a cosmetically acceptable appearance, permitting the patient to more comfortably and confidently resume social interactions during the postoperative healing period. This article presents two case reports describing the use of interim facial prostheses to provide rapid patient rehabilitation. PMID- 12567059 TI - Management of breast carcinoma identified intraoperatively during reduction mammaplasty. AB - Before reduction mammaplasty, patients should be assessed for breast cancer risk and evaluated with physical examination and mammography. Patients determined to be at increased risk should be informed of the possibility of occult breast carcinoma and the potential treatment options. Intraoperative discovery of an occult carcinoma may allow for breast conservation therapy under certain circumstances if an appropriate biopsy sample is taken from the tumor and if clear margins are obtained. PMID- 12567060 TI - Subglanular stricture: rare penile anomaly resulting from circumcision. AB - Circumcision is a commonly performed minor surgical procedure; however, it may lead to serious complications if performed by poorly trained health care providers. In this case report, a subglanular stricture causing a mushroom-like deformity resulting from circumcision is presented. PMID- 12567061 TI - Free anterolateral thigh perforator flap for reconstruction of dystrophic epidermolysis bullosa-associated squamous cell carcinoma in the foot: case report. AB - The authors present a patient with dystrophic epidermolysis bullosa-associated squamous cell carcinoma in the foot. The patient underwent successful reconstruction with a free anterolateral thigh flap based on a perforator with minimal donor site morbidity. PMID- 12567062 TI - Easy method of capturing microscopical images by using a consumer-type digital camera. AB - Taking pictures of microstructures is difficult, requiring sophisticated cameras coupled by the microscopes. Furthermore, it may not be feasible to find an operating microscope paired with a camera, especially in laboratory conditions. Considering the difficulty of obtaining microscopic photographs in clinical and laboratory settings, this report describes a practical method of digital photography of microstructures using a consumer-type digital camera. The technique can be summarized simply as placing the lens of a consumer-type digital camera over the eyepieces of a surgical microscope, whereby the anatomic structure of interest is focused. The image thus obtained is transferred to the computer, using easily available software programs. The authors conclude that this method is an inexpensive and practical way of capturing photographs of microstructures, obviating the need for using microscopes coupled by the cameras. PMID- 12567063 TI - Rotation flap of the anterior rectus abdominis sheath for hernia prevention in TRAM breast reconstruction. AB - Prevention of hernia or bulge of the abdominal wall after TRAM breast reconstruction has been a challenge for the reconstruction surgeon. Different techniques have been described to avoid this complication. The use of anterior rectus abdominis sheath (ARAS) for the repair of various abdominal wall hernias has been well described in the literature and is the basis of the authors' technique. The authors present the use of ARAS flap in TRAM breast reconstruction. It is a simple and safe technique using autologous tissues for hernia or bulge prevention. PMID- 12567064 TI - Modification of split-thickness skin graft: cosmetic donor site and better recipient site. AB - The author reports an alternative method of caring for the donor sites of split thickness skin graft that involves the immediate return of superficial thin-layer split skin graft to the donor sites while using the deep split-thickness skin graft for replacement over the defects. The proposed method would decrease the morbidity associated with other established techniques of donor site management, including prolonged healing time, fluid loss, discomfort, pain, and undesirable cosmetic results. As for the recipient site, satisfactory cosmetic effects and less pigmentation changes could be achieved with this type of simple modification of a laminated graft. PMID- 12567065 TI - Future of fat as raw material for tissue regeneration. AB - Tissue replacement traditionally requires use of autologous tissue and is associated with the attendant morbidity of donor site harvest. In the case of allograft transplantation, there are concerns, similar to those associated with organ transplantation, of rejection and immunosuppression. For these reasons, emphasis has been placed on the development of tissue-engineered substitutes that incorporate autologous stem cells into tissue-engineered scaffolds. The authors' laboratory has characterized a population of cells obtained from processed lipoaspirate (PLA), which have the capacity in vitro to differentiate into osteoblasts, chondrocytes, myocytes, adipocytes, and neuron-like cells. Adipose tissue is an abundant, expendable, and easily obtained tissue that may prove to be an ideal source of autologous stem cells for engineering tissues. PMID- 12567066 TI - Lifelong learner/lifelong teacher. PMID- 12567067 TI - Treatment of burn scars with the erbium:YAG laser. PMID- 12567068 TI - The long-term evaluation of tracheostomy in the management of severe obstructive sleep apnea. AB - OBJECTIVE: To determine the long-term success and morbidity of tracheostomies for patients with severe obstructive sleep apnea. STUDY DESIGN: A retrospective study of patients who have undergone tracheostomy for documented obstructive sleep apnea at Hennepin County Medical Center since 1979. METHODS: Retrospective chart review of all tracheostomy patients operated January 1, 1979, to June 30, 1999, were evaluated. Follow-up data from patient records, death certificates, and DMV. RESULTS: Seventy-nine patients (70 men and 9 women; age range, 25-70 y; mean age, 47 y) received tracheostomies at a regional sleep disorder center. Respiratory distress index ranged from 45 to 146 (mean value, 81). Tracheostomy eliminated obstructive sleep apnea in all cases. Follow-up ranged from 3 months to 20 years (mean period, 8.3 y). In all, 16 patients had decannulation (range, 2 mo to 13 y): 5 of the patients chose continuous positive airway pressure, 3 grew intolerant of their tracheostomy, 3 had obstructive sleep apnea fully resolve after uvulopalatopharyngoplasty, 3 unknown diagnosis for decannulation, 2 had significant weight loss. Morbidity seen within the first year was primarily limited to granulation tissue, infection, and stoma revision. Fourteen deaths were identified. Average age at time of death was 62 years. Five deaths were cardiopulmonary related, four were from cancer, two were from postoperative complications of unrelated surgery, and one was from aspiration. Tracheostomy related mortality included one postoperative myocardial infarction and one tracheal-innominate fistula. CONCLUSION: Severe obstructive sleep apnea and its comorbid conditions are effectively treated in the long term with tracheostomy. Initial management of more frequent complication is well tolerated. Significant morbidity and mortality are low. Chances of obstructive sleep apnea resolution allowing decannulation remain poor. PMID- 12567069 TI - Paranasal sinus development: a radiographic study. AB - OBJECTIVE: To demonstrate the development of the paranasal sinuses in a pediatric population by computed tomography scans. STUDY DESIGN: Radiology records at a tertiary care institution were reviewed for the computed tomography scans of the face, orbit, or paranasal sinuses in patients aged 0 to 12 years. METHODS: Computed tomography scans were reviewed by a head and neck radiologist and otolaryngologist for the development of the frontal, maxillary, ethmoid, and sphenoid sinuses. The size of the pneumatized paranasal sinuses was measured in two planes and graded on a scale of 0 to 3. Ossification of the maxillary crest and vomer, obliteration of the foramen cecum, and development of agger nasi cells, Haller cells, and the superior turbinate were studied. Patients with syndromes, nasal stenosis, choanal atresia, or cystic fibrosis were excluded from the study. RESULTS: In all, 91 computed tomography scans in 66 patients were studied. Serial development could be followed in 16 patients who underwent repeat scans. Patients were divided into six age cohorts based on their age at the time of the scan: 0 to 3 months (10%), 3 to 12 months (13%), 1 to 3 years (13%), 3 to 5 years (20%), 5 to 8 years (29%), and 8 to 12 years (16%). Ethmoid sinuses were the first to fully develop, followed sequentially by maxillary, sphenoid, and frontal sinuses. Each sinus has a rapid rate of development during specified age cohorts. CONCLUSION: The results will aid the physician when correlating the clinical and radiographic findings of pediatric patients aged 0 to 12 years who are being evaluated for sinus disease and potential surgical intervention. PMID- 12567070 TI - Eosinophilic fungal rhinosinusitis of the lacrimal sac. AB - OBJECTIVES/HYPOTHESIS: Chronic sinusitis is a condition affecting millions of individuals each year. Recent findings indicate that chronic rhinosinusitis is a response to fungi mediated by the eosinophil in some of these individuals. We report a case of eosinophilic fungal rhinosinusitis of the lacrimal sac, an entity not previously reported. STUDY DESIGN: Case report. METHODS: A 69-year-old man presented with a 1-cm cystic lesion in the right-side medial canthal region. This lesion was fluctuant, and mucoid material was freely expressed from the medial canthal area. Endoscopic examination revealed bilateral nasal polyps anteriorly and superiorly with mucopurulent drainage from both maxillary sinuses. He had undergone three prior sinus surgeries and two prior lacrimal cannulations elsewhere. A computed tomography scan showed opacification of both maxillary sinuses, ethmoid sinuses, and the left sphenoid sinus and moderate thickening in the frontal sinuses. There was significant soft tissue prominence in the area of both lacrimal fossae with erosion into the medial orbit. The patient underwent bilateral endoscopic revision ethmoidectomies, middle meatal antrostomies, sphenoidotomies, frontal sinusotomies, and endoscopic right-side dacryocystorhinostomy at the Department of Otorhinolaryngology, Mayo Clinic (Rochester, MN). RESULTS: At the time of surgery, massive amounts of thick mucoid material were aspirated from both lacrimal regions. The aspiration of the lacrimal duct on the left side was sufficient treatment, whereas the dacryocystorhinostomy was necessary to decompress the cyst of the face on the right side. The material aspirated was allergic mucin, and the patient met the criteria for diagnosing eosinophilic fungal rhinosinusitis. The patient was started on a regimen of antifungal nasal irrigations postoperatively and has done well in follow-up. CONCLUSIONS: Although never previously reported, eosinophilic fungal rhinosinusitis can occur in the lacrimal sac. It is important to recognize this so that the underlying disease process can be treated appropriately. PMID- 12567071 TI - Quality of life assessment in nonmelanoma cervicofacial skin cancer. AB - OBJECTIVES/HYPOTHESIS: Health-related quality of life (QOL) assessment of patients with nonmelanoma skin cancer is poorly understood. The objectives of the study were to determine the general QOL of patients with cervicofacial skin cancer and to identify patient, clinical, and preventive behavior variables associated with patients' QOL. STUDY DESIGN: Cross-sectional study of 121 consecutive patients (65 female and 56 male patients) presenting to a dermatological Mohs surgery clinic with nonmelanoma skin cancer of the head and neck. METHODS: Quality of life assessment was performed before counseling or treatment. Measures included the Medical Outcomes Study Short Form 36-item Health Survey (SF-36) and the Functional Assessment of Cancer Therapy-General (FACT-G). RESULTS: Both instruments demonstrated good internal consistency as measured by Cronbach's alpha (SF-36, alpha = 0.45-0.91; FACT-G, alpha = 0.61-0.90). The SF-36 scores were similar to historical norms. Bivariate analysis indicated significant correlation coefficients between QOL and patients' coexisting illnesses and medical risk factors. Sun-protective behaviors were associated with better QOL. The relationship appeared to be minimally influenced by patients' sociodemographic characteristics and disease-related variables (size, location, extent). CONCLUSIONS: Sun-protective behaviors were positively associated with certain QOL subscale scores in the population in the study. General QOL instruments demonstrated minimal impact of nonmelanoma skin cancer on patients at initial diagnosis. However, general measures may not be sensitive to the impact of nonmelanoma skin cancer. The development of a more disease-specific instrument may be necessary to evaluate this disease process. PMID- 12567072 TI - Laryngopharyngeal dysfunction from the implant vagal nerve stimulator. AB - OBJECTIVES/HYPOTHESIS: The objective of the study was to examine the side-effect profile of the vagal nerve stimulator. Vagal nerve stimulators have been used to treat intractable seizures in all age groups. They provide relief to the patient with a seizure disorder by decreasing the overall number and severity of seizure activities. Although significant complications are rare, many patients have some complaint, usually of their voice. STUDY DESIGN: A retrospective evaluation of four patients with intractable epilepsy. METHODS: Evaluation of charts and medical records and endoscopic examination of the larynx. RESULTS: In this small series, all four patients had implantation-related paresis. Three of the four appear to have side effects from device activation. CONCLUSIONS: Patients in whom a vagal nerve stimulator is placed can have adverse side effects. These can be related to the surgical manipulation of the vagus nerve, resulting in a temporary paresis of the vocal folds. A second set of side effects is related to the actual electrical stimulation of the device, and these side effects can directly affect the laryngeal musculature. PMID- 12567073 TI - Bordetella pertussis causing otitis media: a case report. AB - OBJECTIVE/HYPOTHESIS: is an unusual cause of otitis media with effusion. We report a case in a neonate. STUDY DESIGN: Case report. METHODS: A case is presented. RESULTS: A 1-month-old infant girl was admitted because of respiratory distress and subsequently was found to have pertussis. On examination she was noted to have a middle ear effusion. Myringotomy was performed for culture. Culture confirmed. We were unable to find a previous report of otitis media. CONCLUSION: has been identified in the middle ear effusion of an infant with the pertussis syndrome. PMID- 12567074 TI - Effects of steel scalpel, ultrasonic scalpel, CO2 laser, and monopolar and bipolar electrosurgery on wound healing in guinea pig oral mucosa. AB - OBJECTIVE: The study's objective was to compare instrument performance and tissue healing when steel scalpel, ultrasonic scalpel, monopolar or bipolar electrosurgical instruments, or CO2 laser was used in an animal oral surgery model. STUDY DESIGN: Prospective, blinded, randomized. METHODS: Adult guinea pigs (N = 70) were randomly assigned to 5 groups (14 animals per group) for excision of 2-cm, full-thickness oral mucosa using steel scalpel, ultrasonic scalpel, monopolar or bipolar electrosurgical instruments, or CO2 laser. Postoperative pain was measured indirectly using weekly body weight changes. Animals from each group were killed on days 0, 7, 14, 21, and 28. Specimens were harvested for blinded histopathological study and tensile strength measurement. Instrument performance (hemostasis, tissue coagulation, tissue sticking) and wound healing (tissue re-epithelialization, degree of inflammation) were primary outcomes. Statistical analysis was performed using analysis of variance. RESULTS: The ultrasonic scalpel was the best tool in controlling hemostasis, tissue coagulation, and tissue sticking. Significantly higher body weight gain ( P<.05) was noted at day 7 for monopolar and CO2 laser groups. Greatest tensile strength was seen in the steel scalpel and ultrasonic scalpel groups at the end of 28 days. Tissue re-epithelialization was fastest for the steel scalpel and ultrasonic scalpel groups (complete by day 7). Complete re-epithelialization of wounds of all treatment groups occurred by day 28. All groups had acute inflammation. Complete resolution of inflammation by day 14 took place in the steel scalpel and ultrasonic scalpel groups only. CONCLUSION: Use of the ultrasonic scalpel produced faster re-epithelialization and greater tensile strength than laser or electrosurgical instruments, with results comparable to those seen with the steel scalpel. PMID- 12567075 TI - The effect of mitomycin on extracellular matrix proteins in a rat wound model. AB - OBJECTIVE: To evaluate the effect of topical and injected mitomycin on the expression of extracellular matrix proteins by fibroblasts in an early surgical wound model. STUDY DESIGN: A prospective, controlled study in a rat wound model. METHODS: Six linear incisions were placed on the backs of each of three Sprague Dawley rats, and polyvinyl alcohol sponges were implanted. Two control wounds were implanted with saline-soaked sponges. The two topical test group wounds were treated with 0.5 mg/mL topical mitomycin for 2 minutes, followed by sponge implantation. The two injection test group wounds were injected with 0.3 mL mitomycin (0.5 mg/mL) before incision and sponge implantation. Each incision was closed uniformly with 3-0 nylon suture. The sponges were harvested on the tenth postoperative day. Fibroblasts that had grown into the sponges were separated, and polymerase chain reaction analysis was used to quantify the expression of messenger RNA for several extracellular matrix proteins. RESULTS: The expression of mRNA for some extracellular matrix proteins (elastase, hyaluronidase, and procollagen) was downregulated in the mitomycin test groups. The effect was more pronounced in the topical mitomycin test group compared with the injection test group. The wounds in the topical group were prone to dehiscence, and the wounds in the injection group demonstrated poor healing when compared with controls. CONCLUSIONS: Mitomycin may inhibit wound healing by downregulating the gene expression for extracellular matrix proteins. This effect may be selective and may be more pronounced on inducible genes. Such findings prompt further studies regarding possible "best time" windows and selective gene suppression. The use of mitomycin may be limited in situations where wound integrity is necessary. PMID- 12567077 TI - Healing of composite chondrocutaneous auricular grafts covered by skin flaps in nasal reconstructive surgery. AB - OBJECTIVE/HYPOTHESIS: Healing of composite chondrocutaneous auricular grafts combined with overlying skin flaps for reconstruction of the internal lining in partial nasal defects should be evaluated. STUDY DESIGN: A retrospective chart review and prospective clinical follow-up of 15 patients with partial nasal defects. METHODS: Fifteen patients with partial defects of the middle and lower third of the nose were reviewed. Nineteen composite ear grafts were used for reconstruction of the internal nasal lining in the patient group. The composite grafts were used in combination with overlying nasolabial transposition flaps. In some patients paramedian forehead or turn-in flaps were needed as well. Functional and aesthetic results of nasal reconstruction were subjectively graded by the patients and medical doctors of the department. RESULTS: Three of 19 transplanted composite grafts showed postoperative deformation. Relative shrinkage was observed in 5 of 19 composite grafts, whereas complete loss of the composite graft was seen in only one patient. Nasal airway obstruction was observed in the patients with deformity or loss of composite graft (4 of 15 patients), and an unsuccessful aesthetic result was seen only in the patient with total composite graft failure (1 of 19 transplanted grafts). CONCLUSIONS: Composite ear grafts may provide good functional and aesthetic results when combined with overlying skin flaps in reconstruction of partial nasal defects. Even though composite ear grafts used for restoration of the inner nasal lining may be deformed or absorbed and impairment of nasal airway patency or aesthetic result may be postoperatively observed, the combination of composite ear grafts with locoregional transposition flaps provides additional reconstructive options for selected nasal defects. PMID- 12567076 TI - Interleukin-1 facilitates airway epithelial migration in response to injury. AB - OBJECTIVE: To test the hypothesis that interleukin (IL)-1 plays a permissive role in respiratory epithelial cell migration and proliferation. STUDY DESIGN: Primary cultures of porcine respiratory epithelial cells or tracheal organ explants were cultured in the presence or absence of function-blocking antibodies to IL-1. Areas of epithelial cell outgrowth were determined in control and antibody treated organ explants daily for 4 days. At intervals, cultured cells were collected for cell counting and viability determination. Time course and dose response curves were constructed for control and antibody-treated groups. RESULTS: Interleukin-1 secretion into culture supernatants increased sharply from days 3 to 7. Outgrowths from tracheal explants were reduced by greater than 60% by single antibody treatment, and by over 90% by treatment with antibodies to both IL-1alpha and -1beta by day 4 of culture. Function-blocking antibodies to IL 1 significantly reduced cell number by day 7 of culture. CONCLUSIONS: Interleukin 1 is produced by respiratory epithelial cells in culture during log phase growth and plays a permissive role in cell migration and proliferation. PMID- 12567078 TI - Choanal atresia: a twenty-year review of medical comorbidities and surgical outcomes. AB - OBJECTIVE: To review medical comorbid conditions and surgical outcomes for children treated for choanal atresia (CA). STUDY DESIGN: Retrospective review (1979-1998). METHODS: Participants included 78 children, aged newborn to 18 years, in the setting of an academic pediatric hospital. Main outcome measures were type and number of procedures and airway patency. RESULTS: Seventy-eight children (48 female and 30 male patients) were treated and followed for CA, with an average follow-up of 35 months. Thirty-five children (45%) had unilateral CA, and 43 children (55%) had bilateral CA. There was no statistically significant sex difference between unilateral and bilateral CA. Concomitant medical problems were common, mostly otitis media with effusion (32%), upper and lower airway diseases (32% and 23%, respectively), cardiac anomalies (19%), and gastrointestinal tract disorders (18%). Statistically significant correlations were found for bilateral CA and cardiac disorders (P =.04), CHARGE syndrome ( P=.002), obstructive sleep apnea ( P=.003), hematological problems (P =.001), and prematurity or failure to thrive (P =.006). Surgery was effective in establishing a patent airway in all cases. Average ages at first surgery were 25.2 months for unilateral and 2.4 months for bilateral CA. Unilateral CA required, on average, 2.7 total procedures, including dilation and removal of stents, compared with bilateral CA, which required 4.9 procedures. Nearly all children underwent correction by means of the transnasal approach. CONCLUSIONS: To the knowledge of the authors, the present report encompasses the largest series of children treated for CA. The high incidence of comorbid conditions mandates thorough medical evaluation, particularly for otological, airway, cardiac, and gastrointestinal tract diseases. PMID- 12567079 TI - Ear, nose and throat disorders in children with Down syndrome. AB - OBJECTIVE: To document the reasons for which children with Down syndrome were referred to a pediatric otolaryngology practice, the underlying causes for these referrals, and the complications of routine surgical therapy. STUDY DESIGN: The study is a retrospective review of children referred to the Pediatric Otolaryngology Clinic at the University of New Mexico Health Sciences Center (Albuquerque, NM) during a period of 2.5 years. METHODS: Data were collected on 55 parameters related to ethnicity, demographics, diagnosis, surgical therapy, complications, and systemic comorbid conditions. RESULTS: The ethnicity of the study population was predominantly Hispanic or Latino (62%). The majority of children (76%) were referred for upper airway obstruction. Obstructive sleep apnea and laryngomalacia were the most common disorders in these children. An otological disorder was diagnosed in 70% of the children. Complications occurred after 27% of procedures for insertion of pressure equalization (PE) tubes to treat recurrent otitis media. Systemic comorbid conditions were present in 93% of the children, and the most common was gastroesophageal reflux disease. CONCLUSIONS: Obstructive sleep apnea and laryngomalacia were the most common reasons for referral of children with Down syndrome. Routine surgical procedures that required general anesthesia caused complications that are not common in other children. Treatment for systemic comorbid conditions should be considered as a component of therapy for otolaryngological disorders in children with Down syndrome. PMID- 12567080 TI - 'Eosinophilic fungal rhinosinusitis': a common disorder in Europe? AB - OBJECTIVES/HYPOTHESIS: The traditional criteria for the diagnosis of allergic fungal sinusitis include chronic rhinosinusitis, "allergic mucin" (mucus containing clusters of eosinophils), and detection of fungi by means of histological examination or culture. In 1999, a group of Mayo Clinic researchers, with a novel method of mucus collection and fungal culturing technique, were able to find fungi in 96% of patients with chronic rhinosinusitis. Immunoglobulin E mediated hypersensitivity to fungal allergens was not evident in the majority of their patients. Because the presence of eosinophils in the allergic mucin, not a type I hypersensitivity, is probably the common denominator in the pathophysiology of allergic fungal sinusitis, the Mayo Clinic group proposed a change in terminology from allergic fungal sinusitis to eosinophilic fungal rhinosinusitis. Using new techniques of culturing fungi from nasal secretion, as well as preservation and histological examination of mucus, we investigated the incidence of "eosinophilic fungal rhinosinusitis" in our patient population. STUDY DESIGN METHODS: In an open prospective study nasal mucus from patients with chronic rhinosinusitis as well as from healthy volunteers was cultured for fungi. In patients, who underwent functional endoscopic sinus surgery, nasal mucus was investigated histologically to detect fungi and eosinophils within the mucus. RESULTS: Fungal cultures were positive in 84 of 92 patients with chronic rhinosinusitis (91.3%). In all, 290 positive cultures grew 33 different genera, with 3.2 species per patient, on average. Fungal cultures from a control group of healthy volunteers yielded positive results in 21 of 23 (91.3%). Histologically, fungal elements were found in 28 of 37 patients (75.5%) and eosinophilic mucin in 35 of 37 patients (94.6%). Neither fungi nor eosinophils were present in 2 of 37 patients (5.4%). CONCLUSIONS: Our data show that the postulated criteria of allergic fungal sinusitis are present in the majority of patients with chronic rhinosinusitis. Either those criteria will be found to be invalid and need to be changed or, indeed, "eosinophilic fungal rhinosinusitis" exists in the majority of patients with chronic rhinosinusitis. Based on our results, fungi and eosinophilic mucin appear to be a standard component of nasal mucus in patients with chronic rhinosinusitis. PMID- 12567081 TI - Bacteria detected by culture and 16S rRNA sequencing in maxillary sinus samples from intensive care unit patients. AB - OBJECTIVES/HYPOTHESIS: In critically ill patients, the occurrence of artificial ventilation-acquired sinus disease is common. A possible sinus bacterial infection is occult and is combined with diagnostic difficulties. Culture as a method has limited capacity to verify the presence of bacteria and leaves unanswered the question of a possible infective agent because bacteria are difficult to grow when killed or suppressed by current antibiotic therapy. Hitherto unidentified micro-organisms are also possible in the microenvironments of maxillary sinuses. STUDY DESIGN: Prospective case series. METHODS: Twenty maxillary sinus samples (17 aspirates and 3 lavages) from nine critically ill patients with possible infectious disease were investigated by broad-range 16S rRNA polymerase chain reaction followed by sequencing. These results were compared with the previous culture results from gingiva (passage route), maxillary sinus absorption, and mucosa samples. RESULTS: The contaminations were rare (2 of 20) and corresponded well to culture results. One previously undiagnosed bacterium was found. Two aspirates were negative on both culture and polymerase chain reaction, whereas the corresponding maxillary sinus mucosal cultures had been positive. CONCLUSIONS: Applying a low-contamination maxillary sinuses sampling technique makes 16S rRNA sequencing clinically useful. The polymerase chain reaction and culture results were generally comparable. However, by polymerase chain reaction, bacteria were found that were missed by culturing. Some aspirates were free of bacterial remnants on 16S rRNA polymerase chain reaction, but corresponding mucosal cultures were positive. This could indicate that infection is induced within the tissue. It also indicates that infectious agents introduced into the sinuses may have routes other than the ostium. Further clinical use of 16S rRNA sequencing is required to enlarge our knowledge in applied microbiology and paranasal sinus disease. PMID- 12567082 TI - Salvage frontal sinus surgery: the endoscopic modified Lothrop procedure. AB - OBJECTIVES/HYPOTHESIS: Until recent years, the osteoplastic flap with frontal sinus obliteration has been the gold standard for recalcitrant frontal sinusitis. The present series evaluated the role of the endoscopic modified Lothrop procedure, which has recently been advocated as an alternative. STUDY DESIGN: Prospective non-randomized interventional case series. METHODS: The study prospectively assessed 83 consecutive patients who underwent endoscopic modified Lothrop procedure. The mean age was 52.4 years (SD = 13.6 y) with a male-to female ratio of 3:1. Patients had a mean of six previous sinus surgical procedures with 17 patients having undergone previous frontal sinus obliteration with mucocele formation. Seventy-six patients (91%) had frontal pain or headache as their primary presenting symptom, with 72 having nasal discharge. There were 14 patients who presented with 17 complications of frontal sinus disease. There were eight erosions of the posterior table of the frontal sinus with extension of the mucocele intracranially, seven orbital complications, and one cerebrospinal fluid leak. Twenty-four patients (30%) had fungus cultured from their sinuses at the time of surgery. RESULTS: Six of the 83 patients (7%) developed frontal ostium stenosis resulting in a 93% primary success rate after an average follow up of 21.9 months (SD = 6.1 mo). These patients all underwent a revision modified Lothrop procedure and had a patent frontal ostium at their last review. Twenty one patients (25%) developed recurrent symptoms, which were managed medically. Of these 21 patients, 9 with previously diagnosed fungal sinusitis developed mucosal changes again in their frontal sinuses, but their ostia have remained patent. Four patients have had recurrent infections in the frontal sinuses, and three patients with aspirin-sensitive asthma and polyps have developed polyps again in their frontal sinuses. Five patients continued to have frontal pain without radiological evidence of further frontal disease. No patients required an osteoplastic flap procedure. CONCLUSION: The endoscopic Lothrop procedure is a successful short-term management option for recalcitrant and complicated frontal sinusitis caused by a wide range of diseases. PMID- 12567083 TI - Investigating the nasal cycle using endoscopy, rhinoresistometry, and acoustic rhinometry. AB - OBJECTIVES: Cyclic congestion and decongestion in the two nasal cavities is seen in connection with the respiratory function of the nose. The turbulent behavior of nasal airflow is a prerequisite for adequate contact of inspired air particles with the mucosa. The aim of this study was to gain insight into this turbulent behavior of nasal airflow during the nasal cycle. METHODS: The nasal cycle in 10 healthy human subjects was investigated using endoscopic imaging, rhinoresistometry, and acoustic rhinometry every 20 minutes over a time period of up to 15 hours. The following parameters were recorded for each nasal cavity: airflow resistance, hydraulic diameter, friction coefficient lambda as an indicator for the wall configuration triggering turbulence, transition from laminar to turbulent flow, and the minimal cross-sectional areas. RESULTS: In addition to the known cyclic change of flow resistance and nasal width, a periodic change in the turbulence behavior was observed. In the resting phase, mainly laminar flow was found. During the working phase, the onset of turbulence occurred already at low flow velocities. The increase of turbulence during the working phase is caused by the increase in cross-sectional area in the anterior cavum due to decongestion of the mucosa of the head of the inferior turbinate and the septal tuberculum. CONCLUSIONS: Rhinoresistometry and acoustic rhinometry complement each other. The combination of the two methods provides insight into the functional changes during the nasal cycle and into nasal physiology in general. The authors therefore advocate a combination of the two methods for functional evaluation of the nasal airway. PMID- 12567084 TI - Acoustic rhinometry and paranasal cavities: a systematic study in box models. AB - OBJECTIVE/HYPOTHESIS: Acoustic rhinometry (AR) is a well-established diagnostic tool in rhinology. The aim of the study was to test the hypothesis that the paranasal sinuses are a main cause for inaccuracy of AR in the posterior part of the nose. STUDY DESIGN: Experimental study to evaluate the influence of simulated paranasal sinus volume and of the contralateral side of the nose on AR measurements in "box models." METHODS: Models were measured with paranasal sinus volume simulated between 0 and 25 mL and with the junction between the models and the paranasal sinuses varying in length and diameter. RESULTS: Moderate but distinct modification of the posterior area-distance curve was found within the models after changing size of the paranasal sinuses and its junction to the cavity. The apparent cross-sectional area (CSA) measured in the posterior cavum decreased with the volume of the paranasal sinuses. This effect was limited by the length and the diameter of the paranasal junction, as well as by the concha. No influence of the contralateral side on AR measurements was seen. CONCLUSIONS: Acoustic rhinometry reveals reproducible measurements up to 4 cm from the nostril that correspond with the actual model CSA. Simulated paranasal sinuses appear to partially contribute to the inaccuracy in the posterior part of the area-distance curve. PMID- 12567085 TI - Value of acoustic rhinometry for measuring nasal valve area. AB - OBJECTIVE: To assess the validity of acoustic rhinometry for measuring nasal valve area in human subjects. STUDY DESIGN: A comprehensive study that compared acoustic rhinometry data with computed tomography findings from scans obtained perpendicular to the acoustic axis and perpendicular to the floor of the nose. METHODS: Fifty nasal passages of 25 healthy adults with no nasal disease were examined by acoustic rhinometry and computed tomography. In each case, the area of the nasal valve as measured by acoustic rhinometry was compared with the area calculations from computed tomography sections taken in two different coronal planes, one perpendicular to the acoustic axis and one perpendicular to the floor of the nose. Computed tomography slices perpendicular to the floor of the nose were obtained at two different locations, a specific distance from the tip of the nose and a specific distance from the anterior nasal spine. RESULTS: There was a significant correlation between the nasal valve areas determined by acoustic rhinometry and computed tomography when imaging was obtained perpendicular to the acoustic axis. In contrast, when scanning was obtained perpendicular to the straight axis of the floor of the nose, the correlations between the acoustic rhinometry and computed tomography data were weak. CONCLUSIONS: When any type of imaging is used for comparison with nasal valve areas determined by acoustic rhinometry, the cross-sections should be perpendicular to the acoustic pathway. The results of the study show that acoustic rhinometry is a valuable method for measuring nasal valve area. PMID- 12567086 TI - The chemotactic behavior of eosinophils in patients with chronic rhinosinusitis. AB - OBJECTIVE: To characterize peripheral eosinophil migration in patients with chronic rhinosinusitis in the presence of nasal mucin and nasal tissue extracts. STUDY DESIGN: Prospective, controlled, ex-vivo. METHODS: Peripheral blood eosinophils, nasal mucin, and nasal tissue were harvested at the time of sinus surgery in 10 patients, as well as obtained in 10 healthy control subjects. Extracts were prepared from nasal mucin and nasal tissue. A modified Boyden chamber was used to study eosinophil migration from both patients and healthy control subjects in the presence of both extracts. RESULTS: Patients with chronic rhinosinusitis and all healthy control subjects demonstrated a concentration dependent increased migration of eosinophils in the presence of both nasal mucin and nasal tissue extracts. The percentage of migration was consistently higher for eosinophils from patients with chronic rhinosinusitis compared with control subjects. The difference attained statistical significance in the presence of 50% tissue extract (median percentage of migration, 23.3% vs. 7.8% [ P=.033]). CONCLUSIONS: Nasal mucin and nasal tissue in chronic rhinosinusitis contains chemoattractants, which can induce active eosinophil migration. The eosinophil migration from patients with chronic rhinosinusitis was consistently higher compared with eosinophils from healthy control subjects. Because the eosinophils were obtained from the peripheral blood, this finding suggests activation of eosinophils in the systemic circulation in chronic rhinosinusitis. PMID- 12567087 TI - Vertigo from herpes zoster oticus: superior or inferior vestibular nerve origin? AB - OBJECTIVE/HYPOTHESIS: This study aims to analyze which division of vestibular nerve in the internal auditory canal is responsible for inducing vertigo in patients with herpes zoster oticus (HZO). METHODS: Eight patients (three men and five women) suffered from auricular vesicles, otalgia, and facial palsy, and five of them also had vertigo. Each patient received a battery of tests, including neurological examination, blood examination, audiometry, caloric test, electronystagmography, and vestibular evoked myogenic potential (VEMP) test. RESULTS: All five HZO patients with vertigo had facial palsy on the lesioned side and spontaneous nystagmus beating toward the healthy side. Absent VEMPs were noted in five patients, absent caloric response was noted in four, and sensorineural HL was noted in three. Compared to another three HZO patients without vertigo, all revealed normal responses in both the caloric test and the VEMP test. On MRI scan, two out of four had abnormal gadolinium enhancement along the nerve segments within the internal auditory canal. Six months after treatment, a follow-up caloric test and VEMP test in these eight patients did not alter the results compared with before treatment. CONCLUSION: The nerve trunks within the internal auditory canal are widely affected in HZO patients with vertigo. Both superior division and inferior division of the vestibular nerve attribute to the vertiginous attack. Further, large numbers of HZO patients undergoing caloric testing and VEMP testing are required to support this tentative conclusion. PMID- 12567088 TI - Sudden deafness and Lyme disease. AB - OBJECTIVES/HYPOTHESIS: Otolaryngological manifestations are common in Lyme disease, affecting up to 75% of patients. One of these symptoms is sudden deafness. Hearing loss has been frequently described in Lyme disease; on the other hand, titers seropositive for, the causal agent of this disease, have been found in almost 20% of cases of sudden deafness. No consensual information exists on the outcome of Borrelia-seropositive patients or on the importance of determining Borrelia antibody titers. The present study aimed to determine the prevalence of seropositivity for Borrelia in sudden deafness, describing clinical characteristics and outcomes. STUDY DESIGN: This was a prospective observational study. METHODS: Forty-seven consecutive patients with sudden deafness were enrolled in the study. Demographic data, the presence of tinnitus and vertigo, and low- and high-frequency pure-tone averages were recorded. The percentage of hearing recovery was determined. Data obtained from Borrelia-seropositive patients were described and compared with those from the seronegative group. RESULTS: Titers positive for antibodies were present in 21.3% of the cases. Seropositive and seronegative groups of patients were homogeneous concerning age, sex distribution, the presence of tinnitus and vertigo, and high- and low frequency hearing thresholds. Hearing outcome was not significantly different between the groups of patients. CONCLUSIONS: No distinctive clinical characteristic was found between seropositive and seronegative subjects. The hearing outcome of treated Borrelia-seropositive patients was similar to that of the seronegative group. PMID- 12567089 TI - Vestibular neuronitis in pilots: follow-up results and implications for flight safety. AB - OBJECTIVES To report our experience over the past 12 years with the evaluation and follow-up of pilots with vestibular neuronitis and to discuss points relevant to flight safety and the resumption of flying duties. STUDY DESIGN A retrospective, consecutive case series.METHODS Eighteen military pilots with vestibular neuronitis were examined and followed up. A complete otoneurological workup was performed, including both physical examination and laboratory evaluation. The latter included electro-oculography (EOG) and a rotatory chair test using the smooth harmonic acceleration protocol. RESULTS The mean patient age was 35 +/- 6 years (range, 23 to 42 y), and the average follow-up period was 20.5 +/- 12.8 months (mean +/- standard deviation [SD]; (range, 11 to 48 mo). Electro-oculography caloric test on presentation documented significant unilateral hypofunction in all patients. Thirteen of the 18 patients (72%) had abnormal smooth harmonic acceleration test results. None of the pilots reported any symptoms on follow-up. However, five (28%) had positive otoneurological examination findings, and eight (44%) still had significant caloric lateralization (>25%). The average caloric hypofunction was reduced from 67.8% +/ 29.3% at onset to 40% +/- 16% (mean +/- SD, <.05, paired test). Seven of the patients (39%) had additional electro-oculography findings beyond caloric hypofunction. These included spontaneous, positional, and positioning nystagmus. Smooth harmonic acceleration disease on follow-up was documented in eight patients (44%), five of whom had canal paresis. Eleven patients (61%) demonstrated residual vestibular damage on follow-up. In 6 of these 11 cases (55%), the laboratory evaluation revealed vestibular deficits otherwise undiagnosed by the bedside test battery. CONCLUSIONS The vestibular system plays a central role in orientation awareness and is often challenged by flying conditions. The finding that approximately 60% of pilots who have had vestibular neuronitis continue to show signs of vestibular malfunction, despite apparent clinical recovery, emphasizes the need for a complete vestibular evaluation, including specific bedside testing and laboratory examinations, before flying duties can be resumed. PMID- 12567090 TI - Antigenic as well as nonantigenic stimuli induce similar middle ear responses in the rat. AB - OBJECTIVES/HYPOTHESIS: The observation that during otitis media many different types of micro-organisms have been cultured from effusions indicate that, once present in the middle ear cavity, most types of micro-organisms are able to trigger an inflammatory reaction leading to otitis media. The present study was designed to determine the middle ear response after injection of different substances into the middle ear cavity. STUDY DESIGN: To determine whether and to what extent an inflammatory response of the middle ear depends on the entering agent, the response in the tympanic cavity was studied by otomicroscopy and histological examination after inoculation of various substances. METHODS: Lewis rats were inoculated in transtympanic fashion either with live or heat-killed bacteria (pathogenic and nonpathogenic), Keyhole limpet hemocyanin, active charcoal, or saline. The mucosal response of the challenged middle ears was studied histologically. RESULTS: Irrespective of the inoculated substance, no essential differences in the mucosal response were found. The intensity of the inflammatory response was greater when live bacteria were inoculated. CONCLUSIONS: The present study demonstrates that any substance reaching the middle ear cavity is likely to induce otitis media. These observations emphasize the role of the eustachian tube as "porte d'entree" in the pathogenesis of this disorder. Determination of specific aspects of the eustachian tube involved in protection or in facilitating bacterial translocation will be important for the understanding of the pathogenesis of otitis media and the subsequent development of new therapeutic strategies. In addition, elucidation of bacterial factors involved in the process of colonization and translocation will be of equal importance. PMID- 12567091 TI - Aggressiveness and quantification of epithelial proliferation of middle ear cholesteatoma by MIB1. AB - OBJECTIVE: To assess an easy method that predicts cholesteatoma aggressiveness. STUDY DESIGN: An experimental prospective study. METHODS: Monoclonal antibody MIB1 was used to determine epithelium proliferation in 91 cholesteatomatous ears. Clinical and surgical parameters were compared with proliferation activity to determine pathological and clinical correlation. RESULTS: Statistical correlations were established between hyperproliferation of the cholesteatoma and severe bone erosion (leading to major cholesteatoma complications) and between hyperproliferation and middle ear inflammation (associated with more surgical difficulties and a higher risk of recurrence). A high proliferation index was also found in children's cholesteatoma, which is known to have more aggressive behavior. CONCLUSION: Immunohistochemical use of the MIB1 antibody is a simple technique that can help to determine the aggressiveness of a cholesteatoma. PMID- 12567092 TI - Petrous anatomy for middle fossa approach. AB - OBJECTIVE: The objective was to describe the relationship of anatomical landmarks required for the middle fossa approach to lesions of the petrous apex and internal auditory canal (IAC). Landmarks for safe identification of the IAC are defined, as are two zones (safe zones I and II) anterior and posterior to the IAC. STUDY DESIGN: Temporal bone anatomical study. METHODS: Ten temporal bones underwent high-resolution computed tomography followed by wet bench dissection. A set of 39 different distances between vital structures was measured, where possible, from both radiology films and under the microscope. Mean, median, minimum, and maximum measurements of all distances were determined. Angular measurements were made using a specially designed instrument. RESULTS: Measurements were made in an anatomical position to mimic the middle fossa surgical technique. Measurements are presented for the superior surface of the temporal bone from the sigmoid sinus and inner table to vital structures, from the anterior and posterior petrous apex, and from the IAC. CONCLUSIONS: Petrous apex anatomical knowledge is required for the safe middle fossa approach to the IAC and petrous apex. Two safe zones are defined that should prevent damage to the cochlea and the superior semicircular canal. PMID- 12567093 TI - Gentamicin tympanoclysis: effects on the labyrinthine sensory cells. AB - OBJECTIVES: The objective of the study was to determine whether a selective vestibular hair cell toxicity with sparing of the cochlear hair cells could be achieved by infusing different concentrations of gentamicin into the middle ears of adult cats. STUDY DESIGN: Prospective experimental animal study treating only the left ear of each cat, the right ear serving as individual control. METHODS: Gentamicin solution at concentrations of either 30 or 3 mg/mL was infused daily into the left middle ear of adult cats until overt ataxia occurred. After 1 month or 6 months, each cat was killed and its temporal bones prepared for optical microscopy. RESULTS: Animals treated with 30 mg/mL gentamicin until ataxic required a median of five daily doses. These animals had clear-cut cochlear basal turn hair cell losses accompanying toxic lesions in the utricle and cristae. In contrast, animals treated with 3 mg/mL gentamicin until ataxic required an average of 19 daily doses. These animals had lesions restricted to the utricle and cristae with sparing of the cochlea hair cells. Animals that failed to develop ataxia manifested neither lesions of the cochlear nor vestibular hair cells. CONCLUSION: Gentamicin tympanoclysis in the cat animal model, using a dilute solution and continued once daily until clinical ataxia occurs, is capable of producing selective vestibular hair cell toxicity while sparing cochlea hair cells. PMID- 12567094 TI - Laryngopharyngeal reflux identified using a new catheter design: defining normal values and excluding artifacts. AB - OBJECTIVE: To define normal values for laryngopharyngeal reflux using an improved catheter design with adjustable electrode placement for pH monitoring. STUDY DESIGN: Cohort study of normal volunteers. METHODS: The setting was an institutional-based gastroenterology practice. Subjects included 20 healthy volunteers with no history of laryngeal, pharyngeal, or reflux symptoms. These included 10 men and 10 women (mean age, 33 y; age range, 26-49 y). Ambulatory 24 hour triple-electrode monitoring of pH in distal esophagus, proximal esophagus, and pharynx using new bifurcated probe was performed. The distal electrode was placed 5 cm above the lower esophageal sphincter, with the proximal two electrodes straddling the upper sphincter. The main outcome measure was the number of true pharyngeal reflux episodes. RESULTS: Sixteen of 20 subjects had no episodes, and 2 subjects had only one episode of pharyngeal reflux. The two subjects exceeding this value (8 and 15 episodes, respectively) had abnormal distal and proximal esophageal reflux. Artifacts for pharyngeal reflux due to acidic meals or "pseudoreflux" were excluded. Subject tolerance of this new probe was excellent. CONCLUSION: The triple-electrode bifurcated adjustable pH probe provides a well-tolerated technique to identify true hypopharyngeal acid reflux episodes. When artifacts produced by meals and pseudoreflux are excluded, 90% of normal subjects show no episodes or a single episode over a 24-hour period. PMID- 12567095 TI - Recurrent laryngeal nerve monitoring in thyroid and parathyroid surgery: the University of Michigan experience. AB - OBJECTIVES/HYPOTHESIS: Intraoperative monitoring of the recurrent laryngeal nerve (RLN) is finding increasing acceptance during thyroidectomy. Recently, a laryngeal surface electrode was introduced to enable another form of noninvasive monitoring of the RLN. The present report examines the University of Michigan experience with RLN monitoring using the postcricoid surface electrode. STUDY DESIGN: All patients undergoing partial or total thyroidectomy or parathyroidectomy from January 1999 to July 2001 were considered candidates for the study. Audiologists trained in intraoperative electrophysiological techniques performed all of the monitoring. METHODS: Data collected on each patient included 1) stimulation threshold for a laryngeal compound muscle action potential on initial RLN identification, 2) stimulation threshold of the laryngeal compound muscle action potential on completion of the procedure, and 3) flexible fiberoptic evaluation of the larynx at the initial postoperative visit and at the 3-month follow-up visit. The average duration of follow-up was 9.8 months with a range of 3 to 60 months. RESULTS: The average minimum current required for stimulation on first identification of all nerves was 0.57 mA (+/-0.48 mA). After completion of the procedure a mean threshold level of 0.42 mA (+/-0.55 mA) was obtained during direct RLN stimulation. Post-dissection stimulation of the RLN on the side of tumor dissection was 0.92 mA (+/-0.65 mA) compared with a stimulation threshold of 0.76 mA (+/-0.57 mA) for the nontumor side. CONCLUSIONS: Electromyographic monitoring of the RLN using a postcricoid surface electrode provides a safe, simple, and effective method for intraoperative monitoring during thyroid or parathyroid surgery. Further, evoked electromyography confirms RLN integrity at the conclusion of surgery. PMID- 12567096 TI - Effects of uvulopalatopharyngoplasty on collapsibility of the retropalatal airway in patients with obstructive sleep apnea. AB - OBJECTIVES/HYPOTHESIS: Uvulopalatopharyngoplasty improves sleep-disordered breathing, particularly in patients with sleep-disordered breathing with abnormally high collapsible airway exclusively at the retropalatal airway, as was previously reported. The present study examined the direct and long-term effects of uvulopalatopharyngoplasty on retropalatal airway collapsibility. STUDY DESIGN: Prospective longitudinal study of 18 patients with abnormally high collapsible airway exclusively at the retropalatal airway. METHODS: Closing pressure of the retropalatal airway was estimated by endoscopically obtaining static pressure/area relationship of the passive pharynx in completely paralyzed and anesthetized patients with sleep-disordered breathing (n = 18) before and 3 months after uvulopalatopharyngoplasty. The measurements were repeated 1 year after uvulopalatopharyngoplasty in eight of these patients. RESULTS: Uvulopalatopharyngoplasty decreased retropalatal airway closing pressure by 3.5 cm H2O; furthermore, a direct correlation between the severity of sleep disordered breathing (as determined by nocturnal oximetry) and retropalatal airway closing pressure was revealed. Uvulopalatopharyngoplasty failures revealed retropalatal airway closing pressure greater than atmospheric pressure. Reduced retropalatal airway collapsibility was maintained up to 1 year after uvulopalatopharyngoplasty. Two patients developed marked stenosis of the retropalatal airway with aggravation of sleep-disordered breathing after initial improvement of sleep-disordered breathing and retropalatal airway collapsibility. CONCLUSIONS: Uvulopalatopharyngoplasty decreases the retropalatal airway collapsibility. The effect is maintained for at least 1 year after uvulopalatopharyngoplasty, whereas a few patients develop retropalatal airway stenosis leading to recurrence of sleep-disordered breathing. PMID- 12567097 TI - A matched survival analysis for squamous cell carcinoma of the head and neck in the elderly. AB - OBJECTIVE: To determine whether elderly patients manifest poorer survivals for head and neck squamous cell carcinoma. METHODS: Cases of squamous cell carcinoma of the glottic larynx, oral tongue, and tonsil were extracted from the Surveillance, Epidemiology and End Results database for 1988 to 1998. For each primary site, patients were segregated into two age groups: 50 to 69 years of age and 70 years of age and older. For each case in the latter, elderly group, a reference group case was randomly matched for gender, year of diagnosis, cancer stage, extent of surgery, and radiation therapy. Overall survival and disease specific survival were compared between the two groups with stage stratification with the Kaplan-Meier method. RESULTS: Cases of glottic carcinoma (1882), tongue carcinoma (426), and tonsillar carcinoma (200) in elderly patients were matched to the reference group. Overall mean survival differences were significant for glottic carcinoma (73.9 vs. 96.7 mo, elderly and younger groups, respectively) (P <.001) and tongue carcinoma (59.5 vs. 73.1 mo) ( P=.002) but not for tonsillar carcinoma (46.0 vs. 54.4 mo) ( P=.220). Disease-specific survival differences were significant but small in magnitude for glottic carcinoma (105.9 vs. 114.1 mo, respectively (P <.001) and tongue carcinoma (81.6 vs. 93.6 mo) (P =.009) but not for tonsillar carcinoma (71.5 vs. 70.7 mo) (P =.422). However, after stage stratification, elderly patients often did not exhibit statistically or practically significant poorer overall or disease-specific survivals. CONCLUSIONS: Elderly patients do not necessarily exhibit clinically significant poorer survivals for head and neck squamous cell carcinoma. Age alone should not be used to determine treatment options for elderly patients with squamous cell carcinoma. PMID- 12567099 TI - The anatomical basis for mandibulotomy: midline versus paramidline. AB - OBJECTIVE: To provide an anatomical basis for mandibulotomy. STUDY DESIGN: Prospective. METHODS: The angles between the long axis of the two mandibular central incisors and between the lateral incisor and canine and the corresponding root length were measured in panoramic films of 50 Taiwanese patients who had oral or oropharyngeal carcinomas. RESULTS: The angle and horizontal distance between the mandibular lateral incisor and canine were significantly wider than between the two central incisors. The root length of the canine measured 16 mm on average. CONCLUSIONS: Paramidline mandibulotomy, which cuts through a wider space between the lateral incisor and canine, preserves dentition and the genioglossus and geniohyoid muscles. It appears to be a more preferable approach than midline mandibulotomy. PMID- 12567098 TI - Total glossectomy without laryngectomy as first-line or salvage therapy. AB - OBJECTIVES/HYPOTHESIS: Advanced carcinoma of the tongue can require total glossectomy. Although radiation therapy is of limited efficacy in T3 and T4 tumors involving the base of the tongue, many surgeons are reluctant to suggest highly mutilating surgery. STUDY DESIGN: Retrospective cohort study. METHODS: We compared early postoperative complications, hospital stay duration, function, and oncological outcomes in patients who underwent total glossectomy without laryngectomy as first-line or salvage therapy. RESULTS: Postoperative course and functional outcomes were similar in the two groups. Overall survival was 32% at 3 years and 21% at 5 years. The risk factors for shorter survival were positive margins of resection (P =.002) and tumor spread into the mandible (P =.04). Salvage surgery was not associated with significantly lower survival (P =.09 [NS]). CONCLUSIONS: Postoperative morbidity and functional outcomes are similar after first-line and salvage total glossectomy without laryngectomy. Local tumor control is the main factor influencing survival. PMID- 12567100 TI - Nasal valve suspension: an improved, simplified technique for nasal valve collapse. PMID- 12567101 TI - Blanket removal of the sublingual gland for treatment of plunging ranula. PMID- 12567102 TI - Use of hydroactive hydrocolloid dressing to prevent intraoperative mechanical spread of tumor cells. PMID- 12567104 TI - Detecting glaucomatous damage with multifocal visual evoked potentials: how can a monocular test work? AB - PURPOSE: To understand and improve the detection of glaucomatous damage with multifocal visual evoked potentials (mfVEP) obtained from single eyes. PATIENTS AND METHODS: Monocular mfVEP recordings were obtained from both eyes of 30 individuals with no known visual abnormalities. The 44.5 degrees -diameter display contained 60 sectors. Probability plots, analogous to the total deviation probability plot of the Humphrey Visual Field Analyzer, were created based on tests that compared each eye (monocular test), or the ratio of the responses from the 2 eyes (interocular test), to group norms. RESULTS: For the monocular test, the number of points exceeding the 5% confidence level was not distributed randomly among individuals or among field locations within an individual. Individuals with small signals (i.e., low signal-to-noise ratios [SNR]) showed too many "abnormal" points, while those with relatively large SNR values showed too few. Reasonably good specificity was obtained by defining an abnormality in terms of a cluster of significant points in the mfVEP probability plot. For the interocular test, the results were close to those expected by chance. CONCLUSIONS: Both monocular and interocular tests will be of value when testing glaucoma patients with the mfVEP technique. The interocular test is a more sensitive indicator of glaucomatous damage when a region when healthy has a large signal (SNR) and damage is largely unilateral, whereas the monocular test will be a more sensitive test when a region when healthy has a small signal (SNR). However, the statistics underlying monocular comparisons of mfVEPs are not simple. To obtain high specificity, criteria based on clusters of points need to be used and norms obtained for every laboratory. PMID- 12567105 TI - Prevalence of glaucoma in treated multibacillary Hansen disease. AB - PURPOSE: To determine the prevalence of glaucoma in a population of patients with multibacillary Hansen disease who had completed treatment. PATIENTS AND METHODS: The authors examined 386 of 446 patients with treated multibacillary Hansen disease residing in a geographically limited area. A complete ophthalmic examination including slit-lamp, applanation tonometry, gonioscopy, ophthalmoscopy, and stereobiomicroscopic examination of the optic disc was performed in all subjects. Glaucoma suspects were invited to the base hospital for further examination including automated perimetry. RESULTS: The overall prevalence of glaucoma was 3.6% (CI 1.9-5.3); 1.3% had primary open-angle glaucoma, 7% were primary angle-closure suspects (occludable angles), 1.8% had primary angle-closure glaucoma, and 0.5% had secondary glaucoma. CONCLUSION: The prevalence of primary glaucoma in patients with treated multibacillary Hansen disease was similar to that in the general population, and secondary glaucoma was rare. PMID- 12567106 TI - Intraocular pressure-induced interlamellar keratitis after LASIK surgery. AB - PURPOSE: To describe a case of interlamellar stromal keratitis induced by increased intraocular pressure (IOP) after LASIK surgery. METHODS: Case report and review of the literature. RESULTS: A 53-year-old white man with a history of treated ocular hypertension underwent uncomplicated LASIK surgery. The postoperative course was complicated by markedly elevated IOP induced by topical corticosteroid drops used to treat what appeared to be diffuse lamellar keratitis. Because IOPs remained uncontrolled despite maximal therapy, topical steroids were discontinued after a total of 9 weeks. The IOP rapidly returned to normal range with complete resolution of the corneal findings. Humphrey visual field analysis, confocal scanning laser imaging of the optic nerve, and stereoscopic disc photographs all demonstrated that significant glaucomatous field loss and optic atrophy developed over this 8-week period. DISCUSSION: The IOP should be immediately evaluated in patients who present with interlamellar stromal keratitis more than 1 week after LASIK. If the IOP is elevated, corticosteroid drops should be discontinued to prevent permanent visual loss. Furthermore, if a glaucoma specialist examines a patient with a history of LASIK and unexplained visual field loss, the medical record should be reviewed to determine if the postoperative course was complicated by this diffuse lamellar keratitis-like phenomenon. PMID- 12567107 TI - Primary congenital glaucoma: a novel single-nucleotide deletion and varying phenotypic expression for the 1,546-1,555dup mutation in the GLC3A (CYP1B1) gene in 2 families of different ethnic origin. AB - PURPOSE: To present new molecular genetic data on primary congenital glaucoma from 2 families, 1 isolated case and 3 familial cases due to mutations in the cytochrome P-450 1B1 (CYP1B1) gene. METHODS: All diagnoses were made by slit-lamp biomicroscopy, gonioscopy, cornea and optic disk measurements, ultrasound biometry, and automated static threshold perimetry where possible. Mutation screening was performed by direct sequence analysis of DNA extracted from peripheral blood of the patients and their relatives. RESULTS: For the isolated case, a child of 4 years, a homozygous nucleotide deletion within a tetrad of cytosines (nt622-625, 622delC) was found leading to a predicted nonsense codon 93 truncating the protein by 450 amino acids. For the familial cases, the 3 affected members showed a homozygous mutation 1,546-1,555dupTCATGCCACC for which 9 healthy relatives proved to be heterozygous. The phenotypic expression of these 3 patients varied widely. CONCLUSION: Our results confirm the crucial role of CYP1B1 mutations for congenital glaucoma. PMID- 12567108 TI - Does a family history of glaucoma affect disease severity at the time of diagnosis? AB - PURPOSE: Progressive glaucomatous optic neuropathy is an asymptomatic process with an insidious onset. Patients who are aware of glaucomatous signs and who suspect that they may have the disease may present earlier. If a person has glaucoma, this may alert his or her other family members to seek assessment and thereby permit earlier diagnosis. The authors sought to determine whether glaucoma patients with a family history of the disease were younger and showed less evidence of glaucomatous optic neuropathy at diagnosis than glaucoma patients without a family history of the disease. PATIENTS AND METHODS: Family history of glaucoma, age at diagnosis, and visual field mean defect within 2 years after diagnosis were recorded in 292 patients with primary open-angle glaucoma. Results were analyzed to compare visual field loss with age and family history. RESULTS: At diagnosis, patients with a family history of glaucoma were younger than those without such a history (mean +/- SD, 58 +/- 12.7 years versus 63 +/- 10.8 years; = 3.68, P<0.001). Patients who were younger than 50 years at the time of diagnosis and had a positive family history were significantly less likely to have a worse visual field than those with a negative family history (OR = 0.3; 95% CI, 0.1-0.6; P<0.001), whereas those aged 50 years or older showed no such correlation (OR = 0.9; 95% CI, 0.7-1.3; P = 0.6). CONCLUSION: A family history of glaucoma was associated with a better visual field at diagnosis in patients younger than 50 years, but not in patients 50 years or older. PMID- 12567109 TI - Hyperhomocystinemia in pseudoexfoliation glaucoma. AB - PURPOSE: To determine the prevalence of hyperhomocystinemia in patients with pseudoexfoliation glaucoma. PATIENTS AND METHODS: This prospective study included 30 patients with glaucoma and 30 age-matched controls with no history of ocular disease who were undergoing routine physical checkups. Plasma homocysteine levels of all the study participants were determined using high-performance liquid chromatography, and values exceeding 15 micromol/L were considered elevated. RESULTS: The mean plasma homocysteine level was 16.80 +/- 3.20 and 12.39 +/- 1.97 micromol/L in glaucoma patients and controls, respectively (P<0.0001). Fifteen glaucoma patients (50%) had hyperhomocystinemia compared with 3 controls (10%) (P=0.0015). CONCLUSION: Hyperhomocystinemia may be associated with pseudoexfoliation glaucoma, which may partially explain the increased risk of vascular diseases among patients with pseudoexfoliation syndrome. PMID- 12567110 TI - Concordance of high-pass resolution perimetry and frequency-doubling technology perimetry results in glaucoma: no support for selective ganglion cell damage. AB - PURPOSE: To compare the results of frequency-doubling technology perimetry (FDT), assumed to test the magnocellular visual pathway, and high-pass resolution perimetry (HRP), assumed to test the parvocellular visual pathway, in patients with ocular hypertension or glaucoma. PATIENTS AND METHODS: Forty-eight consecutive patients with glaucoma or ocular hypertension, covering the entire range of optic nerve function from normal to severely damaged, were examined on the same day using FDT and HRP. RESULTS: There was a linear correlation between both global and local indices in FDT and HRP (r = -0.84 P<0.0001 and r = 0.8 P<0.001, respectively). The HRP and FDT classifications agreed in 32 of the 48 (67%) eyes (Cohen kappa = 0.5). There was no significant difference between the ability of the different techniques to detect abnormality. CONCLUSION: The observations in the present study indicate either that both cell populations are similarly affected by glaucomatous damage or that both methods measure activity in the same cell populations. PMID- 12567111 TI - Retinal nerve fiber layer measurement by optical coherence tomography in glaucoma suspects with short-wavelength perimetry abnormalities. AB - PURPOSE: To compare retinal nerve fiber layer (RNFL) measurements in normal and glaucoma subjects with short-wavelength automatic perimetry (SWAP) abnormalities and in chronic primary open-angle glaucoma (CPOAG) patients using optical coherence tomography (OCT). METHODS: Forty-eight eyes of normal subjects, 34 eyes of glaucoma suspects with SWAP abnormalities, and 38 eyes of early CPOAG subjects were recruited. All normal and glaucoma suspects had normal conventional automated perimetry visual field results. All participants underwent full clinical ophthalmologic evaluation followed by OCT RNFL measurements. RESULTS: Compared with normal controls, OCT RNFL thickness was significantly lower in glaucoma suspects with abnormal SWAP (inferotemporal and superotemporal thickness values) and CPOAG patients (average, superior, inferior, inferotemporal, and superotemporal thickness values) (P < 0.01). Some parameters were found to be significantly lower in CPOAG patients than the glaucoma suspects with abnormal SWAP (average, inferior, inferotemporal, and superotemporal thickness values) (P < 0.01). CONCLUSIONS: OCT RNFL measurements appear to correlate well with SWAP abnormalities in glaucoma, and may detect glaucomatous damage earlier than standard conventional automated perimetry. This study suggests that OCT may recognize the earliest evidence of structure alterations in CPOAG. PMID- 12567112 TI - Readability of ocular medication inserts. AB - OBJECTIVE: To determine the readability of ocular medication inserts and whether they are an appropriate source of medication information for patients. MATERIALS AND METHODS: The Flesch-Kincaid and SMOG readability formulas were used to calculate the readability of 10 common glaucoma medication inserts (Alphagan, Azopt, Betoptic, Betimolol, Cosopt, Optipranolol, Rescula, Trusopt, Timoptic, and Xalatan) and 6 widely used nonglaucoma medication inserts (Alrex, Lotemax, Ocuflox, Patanol, Pred Forte, and Zaditor). RESULTS: The 10 glaucoma medication inserts surveyed required an average overall grade level of 12.9 +/- 0.6 by the Flesch-Kincaid Index and of 13.5 +/- 0.6 by the SMOG formula. The 6 nonglaucoma medication inserts had an overall grade level of 11.1 +/- 0.6 by the Flesch Kincaid Index and of 11.7 +/- 0.9 by the SMOG formula. All medications reviewed were above the eighth-grade level recommended by the Flesch-Kincaid Index for public materials. CONCLUSIONS: Ocular medication inserts are too complex to be an adequate source of medication information for the average American adult. This study highlights the need for improving communication and education regarding patients' medications. PMID- 12567113 TI - The effect of latanoprost on accommodation in young patients with ocular hypertension. AB - PURPOSE: To determine whether topical latanoprost 0.005% had an effect on accommodative capacity after 1 month of treatment in young patients with ocular hypertension. PATIENTS AND METHODS: Fifteen patients (9 men and 6 women, aged 20 40 years) with ocular hypertension were instructed to instill the drug to their right eye for 1 month; the left eye received no medication and served as control. Near point of accommodation, refraction, and intraocular pressure (IOP) were determined after 1 month of treatment. RESULTS: After 1 month of treatment with latanoprost, the mean IOP was reduced by 22.8% (5.8 mm Hg) in the treated eye and 1.86% (0.47 mm Hg) in the untreated control eye. No significant changes in refraction or near point of accommodation were observed in the treated or untreated eye. CONCLUSION: Short-term treatment with latanoprost 0.005% does not effect refraction or accommodation in young patients with ocular hypertension. PMID- 12567114 TI - Subconjunctival injection of autologous platelet concentrate in the treatment of overfiltrating bleb. PMID- 12567115 TI - Early detection of glaucoma. PMID- 12567116 TI - Glial modulation of retinal ganglion cell death in glaucoma. PMID- 12567117 TI - Influence of corneal variables on accuracy of intraocular pressure measurement. PMID- 12567118 TI - Glaucoma in India. PMID- 12567119 TI - Some thoughts on the pressure dependence of uveoscleral flow. PMID- 12567121 TI - The medieval medical third party: compulsory consultation and malpractice insurance. PMID- 12567122 TI - Self-inflicted risks of orthopaedic surgery. AB - The advancements in medicine, including orthopaedic treatment, since 1960 have so greatly improved quality of life that human nature has changed in its social goals. These technological innovations, especially the products created, have resulted in a self-inflicted increase in risk of practice. This risk will not be leveled until society recognizes that it expects that the physician not only be a healer, but able to fulfill the social goals of the patient. PMID- 12567123 TI - Lawyer's advice on physician conduct with malpractice cases. AB - The conduct of the orthopaedic surgeon can directly influence his or her risk of a malpractice lawsuit. Communication and caring for the patient summarize positive conduct. Filing of a lawsuit requires certain procedures and the physician must respond to these correctly. The surgeon must completely educate himself or herself about the case and be prepared by his or her lawyer for the deposition and possible subsequent trial. PMID- 12567124 TI - Malpractice: love thy Patient. AB - The Hippocratic oath commands doctors to be the patient's supreme advocate. Coupling this command with a well-trained physician is the sine qua non of orthopaedic risk management. A favorable doctor and patient relationship also is essential. Avoiding a malpractice lawsuit involves the physician's ability to anticipate and remedy adverse occurrences in diagnosis and treatment, and knowing the essence of informed consent and awareness of the standard of care. Documentation is the common denominator in the defense of malpractice claims and for the surgeons, the perioperative notes are paramount. The occurrence of a malpractice lawsuit is more often the result of the failure to practice the art of medicine by the orthopaedist rather than a failure to apply the science of orthopaedics. PMID- 12567125 TI - A malpractice episode: a sequence of events. AB - A malpractice suit is an event that no orthopaedic surgeon wants to go through; however, reality says you will. The authors outlined a step by step approach to your needs during such a suit and offer possible helpful actions by recommending collecting and securing all of the written documentation, being able to familiarize yourself with the entire events without notes, constant appraisal and discussion of the events with your attorney, considerations for settling, and finally emotional events surrounding the trial. The initial shock and insult eventually will subside, your patients will not note the difference, but you never will forget the event. PMID- 12567126 TI - The Stark II reality. AB - The long awaited final regulations in Phase I of a two-phase rulemaking process under the Stark II law were published on January 4, 2001. The Phase I final rules govern interpretation of the Stark law as it is applied to referrals by a physician for designated categories of health services to entities in which the referring physician has a financial interest. These new regulations are of particular concern to specialists, such as orthopaedic surgeons, whose practices are oriented to ancillary services that are considered designated health services, such as radiology, physical therapy and durable medical equipment, and where the availability of clear guidance is essential to ensure that medically necessary care is provided in a manner that complies with law. However, rather than the "brightline" guidance that the healthcare community sought, the new regulations create uncertainty in areas that had not existed before. The new regulations require physicians to evaluate the full range of their business and professional relationships to avoid the risk of nonpayment of claims, civil money penalties, or program exclusion after the effective date of the new regulations. PMID- 12567127 TI - Pioneering in the first century of hip replacement: experiences of a surgeon designer. AB - The current author offers an anecdotal history of the total joint replacement industry in North America that emerged in the 1960s and of the individuals and organizations that shaped it. The author's career as an arthroplasty pioneer, influences that helped him become interested in uncemented fixation techniques, and his impression of the requirements the industry places on emerging orthopaedic leaders also are chronicled. PMID- 12567128 TI - The risk of product liability: orthopaedic surgeons can be trapped. AB - Orthopaedics has become the specialty that it is because of the technologic advancements of medicine in designing successful implants. However, the use of implants introduces a new risk of product failure. The orthopaedic surgeon needs to be aware of the potential consequences of the risks of use of devices in treating patients. PMID- 12567129 TI - Responsibilities when confronting a defective surgical implant. AB - The senior author shares lessons from his experience with faulty knee implants that resulted in severe polyethylene wear and 51% failure of 73 unicondylar knee arthroplasties within 2 years after implantation. Ensuing research showed that the components had been sterilized by gamma irradiation in air and had shelf lives ranging from 4.5 years to 6.5 years. It was learned that gamma-irradiated components with more than 5-year shelf lives remain in inventories without information showing their sterilization dates. This experience reiterated the importance of staying abreast of the literature and reports concerning products and of being alert to unexpected patient outcomes relatively soon after implantation. If a surgeon suspects a faulty prosthetic device, he or she should contact the manufacturer and the United States Food and Drug Administration, so that an investigation can begin. In such situations, the physician's primary responsibility lies with the patient, and it is necessary to arrange extra time and staff resources to give the affected patients information, compassion, added clinical monitoring, and treatment. Because the potential exists for gamma irradiated in air implants with lengthy shelf lives to remain undetected in inventories, the current authors recommend that surgeons select implants sterilized by a different process. PMID- 12567130 TI - An analysis of orthopaedic liability in the acute care setting. AB - The risk of litigation in the emergency room is not directly proportional to the severity of injury. In fact, the risk of initiation of legal action is greater in cases of relatively modest permanent and partial disability. In these cases, failure to obtain and document informed consent, failure to effectively communicate with the patient and family, and failure to assure followup and prompt assessment of complications are the factors that have most frequently led to litigation. Orthopaedic surgery ranks fifth among medical specialties in the total number of claims filed and also fifth in the cumulative amount of financial payments. Approximately 30% of filed cases ultimately result in some form of financial payment. The driving force behind litigation is financial gain for plaintiff's attorneys and compensation for plaintiffs. The motive is not the improvement of medical delivery systems or individual behavior. Access to useful data of a legal nature is extremely difficult. Only a small number of cases go on to a jury-based decision of public record. It is difficult to get accurate and complete information on the total number of cases filed, cases withdrawn, terms of settlements, and arbitrations because of the confidential nature of the settlement process. PMID- 12567131 TI - Risks of a large group practice: a personal experience. AB - Formation of a large orthopaedic group practice and practicing within this group carries many risks that are not present in a small group practice. The current authors will discuss the complexity of merging groups, forming corporate policy, hiring executive administrative staff, contracting issues with insurance companies, expanding services and developing ancillary income sources, and the complex interpersonal relationships of a large group practice. PMID- 12567132 TI - The risks of small community practice. AB - Small community practice has risks not present in a metropolitan practice. The practice is general orthopaedics and subspecialization does not thrive. The quality of consulting physicians and the community hospital may not be of the level desired. The benefits for the orthopaedic surgeon are the high respect he or she and his or her family enjoy in the community, and the high level of appreciation that the citizens and patients give to the doctor. The risk for malpractice suits is small because of the familiarity of the patients and the doctor. PMID- 12567133 TI - Future practice risks: obstacles with opportunities. AB - The encouragement of a consumer-directed insurance market and the involvement of corporate America have led to a transformation of the healthcare system that will redefine patient expectations and the role of the physician. The expectation of a higher level of customer service, the role of electronic media, and an earnest regard for healthcare by the patient will impact the physician. Consumer-directed healthcare will call for a reprioritization, focusing on service, convenience, competency, quality, and low cost. Indifference to patient courtesies or failure to become involved in modern technology may cause the physician to be bypassed from this new healthcare system. As the business world integrates the business model into the medical profession, healthcare organizations will restructure themselves into well-organized business enterprises employing physicians. The singular physician may no longer be competitive with such business enterprises, leading to consolidation of organizations and a significant decision for the physician. The physician will have the unusual opportunity to either remain exclusively a medical professional or to also venture into the business world as an equity investor, participating in financial growth possibilities of those medical practice organizations that are for-profit. With the emergence of these new equity models of healthcare, the primary focus will be on profit, production, and service efficiencies. It is the responsibility of the physician to maintain professionalism and the commitment to quality and appropriateness of patient care. However, at the same time, the physician has the unique opportunity to participate in the distribution, economics, and subsequent financial profit of healthcare. The medical experience of the physician, coupled with business acumen, will be an unbeatable combination. PMID- 12567134 TI - Milwaukee shoulder: correlating possible etiologic variables. AB - The current study evaluated the relative correlation of apatite crystal-induced inflammation and rotator cuff deficiency in the development of cuff tear arthropathy. Thirty-seven patients with full thickness rotator cuff tears were evaluated by history, physical examination, and plain radiographs. Thirty patients had surgical intervention for their rotator cuff defects, and calipers were used intraoperatively to quantify the size of the tear in its largest diameter. The remaining seven patients were treated nonoperatively and the size of the tear was quantified using magnetic resonance imaging. Synovial fluid was obtained from all patients and analyzed for crystal content using an alizarin red stain. Synovial fluid also was analyzed for leukocyte count and differential, prostaglandin E, and matrix metalloproteinase. An unpaired Student's t test revealed that significantly higher levels of prostaglandin E were found in the synovial fluid of patients with apatite crystals, shown by alizarin red stain. Chi squared analysis showed that patients with elevated crystal levels were significantly more likely to have large rotator cuff tears or glenohumeral arthritis. Establishing such relations potentially can elucidate the etiology and treatment of this complex disorder.(2) (2) PMID- 12567135 TI - Glenoid inclination is associated with full-thickness rotator cuff tears. AB - Anatomic factors, such as a hooked acromion, have been associated with rotator cuff disorders. Orientation of the glenoid relative to the scapula has been implicated in shoulder instability, but it has not been linked to rotator cuff disorders. The purpose of the current study was to test the hypothesis that superior inclination of the glenoid is associated with full-thickness rotator cuff tears. Glenoid inclination angles were measured from 16 shoulder radiographs of a convenience sample of eight cadavers in which one shoulder had an intact rotator cuff and the other shoulder had a full-thickness rotator cuff tear. Glenoid inclination angles for shoulders with rotator cuff tears were compared with contralateral normal shoulders using nonparametric statistical analysis. The glenoid inclination angle was greater in cadaver shoulders having full-thickness rotator cuff tears (98.6 degrees ) than in shoulders without tears (91.0 degrees ). A second experiment was done to assess the reliability of using 34 Grashey view radiographs from a clinical population to measure glenoid inclination angle. A method to measure the glenoid inclination on Grashey views was tested and was found to correlate with the inclination angles measured on cadaveric scapulae. Intrarater reliability of measurements from clinical Grashey views was 0.93, and interrater reliability was at least 0.88. PMID- 12567136 TI - Shoulder muscle reflex latencies under various levels of muscle contraction. AB - Previous research in relaxed muscles shows that muscle reflex latencies are too slow to protect the shoulder. However, during athletic activity when injury occurs, some level of shoulder muscle contraction typically exists. The purpose of the current study was to assess shoulder muscle reflex latencies under various levels of muscle contraction. Seventeen healthy subjects participated. A perturbation consisting of an external rotation collision force to the anterior forearm in a position of apprehension under various levels of muscle contraction (0%, 20%, and 50% of a maximum voluntary muscle contraction) was applied. Muscle reflex latencies were measured as the time from perturbation application to onset of muscle activity. Electromyography measured activity onset of the rotator cuff muscles and the primary humeral movers. During 0%, the latissimus dorsi muscle reflex latency was significantly slower than most other muscles. No difference existed between muscles in the 20% and 50% conditions. For the rotator cuff muscles, the reflex latencies significantly quickened for 20% and 50% compared with the relaxed state (0%). Overall, introducing muscle contraction significantly quickened muscle reflex latencies. These results provide clinicians with a better understanding of the role that these reflexes play in joint stability in a position of injury vulnerability like a position of apprehension. PMID- 12567137 TI - Spine fusion using cell matrix composites enriched in bone marrow-derived cells. AB - Bone marrow-derived cells including osteoblastic progenitors can be concentrated rapidly from bone marrow aspirates using the surface of selected implantable matrices for selective cell attachment. Concentration of cells in this way to produce an enriched cellular composite graft improves graft efficacy. The current study was designed to test the hypothesis that the biologic milieu of a bone marrow clot will significantly improve the efficacy of such a graft. An established posterior spinal fusion model and cancellous bone matrix was used to compare an enriched cellular composite bone graft alone, bone matrix plus bone marrow clot, and an enriched bone matrix composite graft plus bone marrow clot. Union score, quantitative computed tomography, and mechanical testing were used to define outcome. The union score for the enriched bone matrix plus bone marrow clot composite was superior to the enriched bone matrix alone and the bone matrix plus bone marrow clot. The enriched bone matrix plus bone marrow clot composite also was superior to the enriched bone matrix alone in fusion volume and in fusion area. These data confirm that the addition of a bone marrow clot to an enriched cell-matrix composite graft results in significant improvement in graft performance. Enriched composite grafts prepared using this strategy provide a rapid, simple, safe, and inexpensive method for intraoperative concentration and delivery of bone marrow-derived cells and connective tissue progenitors that may improve the outcome of bone grafting. PMID- 12567138 TI - Complications associated with reimplantation after girdlestone arthroplasty. AB - Complications associated with reimplantation of a total hip arthroplasty after resection or Girdlestone arthroplasty for treatment of an infected hip are not well-documented. Forty-four hips in 44 patients with a minimum 2-year followup (range, 2-9 years) from the time of reimplantation total hip arthroplasty were evaluated retrospectively. The average Harris hip score changed from 40 preoperatively to 78 at the latest followup. One patient had persistent infection requiring rerevision surgery, and another patient had rerevision surgery for recurrent dislocation. Complications related to the prosthesis consisted of dislocations in five patients (11.4%), recurrent infection in one patient (2.3%), trochanteric nonunion in four patients (9.1%), hematoma in one patient, heterotopic ossification in one patient, and postoperative wound drainage in one patient. Resection arthroplasty of the hip is highly effective in eradicating infection. In the current series, 97.7% of patients were free of infection at the latest followup. However, with an 11.4% dislocation rate and 39% of patients having a persistent limp, alternative approaches need to be evaluated which might reduce these complications. PMID- 12567139 TI - The influence of resting periods on friction in the artificial hip. AB - Insufficient tribologic performance of total joint components is a major cause of prostheses failure. Wear has been studied intensively using testing machines that apply continuous motions. Human locomotion, however, is not well represented by continuous motions alone. Singular events and resting periods are a substantial part of daily activities. Resting does influence adhesion in the artificial joint with possible effects on friction, wear, and loosening. The current study evaluated the effects of resting on the frictional properties of hip prosthesis components. The activity measurements of 32 patients with artificial hip replacements were analyzed for resting durations of the hip. A pin-on-ball screening device was used to determine friction after characteristic resting periods and during continuous oscillating motion. All common articulation pairings were investigated. Prolonged and frequent resting periods of the hip were found for the patients. Initial friction increased with increasing resting duration for all tested materials (between 41% and 191%). The metal-on-metal articulations showed the highest friction level (0.098 for sliding) and the highest increase (191%) in friction with resting duration (0.285 after resting periods of 60 seconds). A high static frictional moment after resting periods might present a risk for aseptic implant loosening. Therefore, large head diameters of metal-on-metal joints should be used with caution, especially when additional unfavorable risk factors such as obesity, weak bone-implant interface, or high activity level are present. PMID- 12567140 TI - Monopolar thermal treatment of symptomatic anterior cruciate ligament instability. AB - Patients with anterior cruciate ligament instability resulting from incomplete tears or elongation in continuity without ligament detachment historically have been treated conservatively or by graft replacement. The literature is sparse regarding alternative treatments. The current study presents experience using monopolar thermal repair on 28 consecutive knees with partial anterior cruciate ligament tears all symptomatically unstable. All lesions were less than 6 months old (average, 77 days; range, 7-180 days) and with a difference of 6 mm or more (average, 9 mm; range, 6-13 mm) when comparing both knees using KT-1000 evaluation. Incomplete tears of the anterior cruciate ligament were seen at arthroscopic evaluation. The rehabilitation protocol included use of a brace for at least 6 weeks and progressive weightbearing. A 2-year minimum followup (range, 24-35 months) was done in all patients following the International Knee Documentation Committee guidelines. The overall outcome was normal or nearly normal in 96% of the patients. One failure occurred at 8 weeks. Twenty-six knees had a KT-1000 difference between 0 and 2 mm (average, 1.9 mm). Because thermal application causes death to some of the cells directly treated, it should be taken into account in selection and application. Immediately after thermal use, the anterior cruciate ligament, although thicker and tighter, is at first weaker than normal. Rehabilitation and compliance are critical during early ligament healing. This procedure seems to be a reasonable alternative to anterior cruciate ligament grafting in selected patients. PMID- 12567141 TI - Injuries when children reportedly fall from a bed or couch. AB - One-hundred-fifteen patients with orthopaedic injuries resulting from a reported fall from a piece of furniture at home were studied to define the relationship of suspected child abuse associated with this mechanism of injury. One hundred thirteen patients sustained fractures or dislocations and two patients were impaled with a pencil and a needle. Each patient was evaluated by a primary care physician or an emergency room physician and by an orthopaedic surgeon. In six of 115 patients (5%), a treating physician filed a child abuse report. In 109 patients (95%) the purported mechanism of injury was considered sufficient to produce the resulting injury. Two of four children younger than 1 year (50%), four of 83 children 1 to 5 years (5%), and none of the 28 children older than 5 years were investigated for potential child abuse. Orthopaedic injuries reportedly attributable to a child falling from a bed or couch at home usually are accidental unless the child is younger than 1 year. PMID- 12567142 TI - Sacral infiltration in pelvic sarcomas: joint infiltration analysis II. AB - The incidence and characteristics of sacral infiltration in pelvic sarcomas were analyzed. Fifty-one patients with a pelvic sarcoma (chondrosarcoma, 15 patients; Ewing's sarcoma, 23 patients; and osteosarcoma, 13 patients) abutting the sacroiliac joint had surgical treatment. Tumor infiltration into the sacrum was suspected based on preoperative images in 18 patients; 15 of 18 patients had histologic tumor invasion. There was a significant difference of median volume of sarcomas with and without infiltration. One of 23 Ewing's sarcomas, seven of 15 chondrosarcomas, and seven of 13 osteosarcomas penetrated the sacroiliac joint into the sacrum. Logistic regression test showed that diagnosis was the most important factor influencing sacral infiltration. Twelve tumors infiltrated through the posterior part of the joint, two tumors infiltrated through the anterior part, and one large tumor infiltrated through an unknown route. To obtain wide surgical margins, patients at risk (elderly, large tumor, or diagnosis of osteosarcoma or chondrosarcoma) for sacral involvement may require extended internal hemipelvectomy with the medial margin extending into the sacrum. High quality imaging studies should be used to assess the need for transarticular resection. PMID- 12567143 TI - Postoperative progression of pulmonary metastasis in osteosarcoma. AB - Early relapse with distant metastasis often is observed in patients with cancer after resection of the primary tumor. It is considered that resection of the primary tumor induces activation of systemic angiogenesis and enhances progression of remote metastasis. The authors show that resection of the primary osteosarcoma tumor enhances progression of pulmonary metastasis in animal osteosarcoma models. Matrigel plug neovascularization assay revealed that systemic angiogenic activity was elevated after primary tumor removal (tumor intact group, 1.61 +/- 0.21 g/dL; tumor removed group, 4.92 +/- 0.35 g/dL). In addition, serum concentration of the angiogenesis inhibitor, endostatin, decreased significantly after primary tumor removal. Treatment with the antiangiogenic reagent TNP-470 suppressed postoperative progression of pulmonary metastasis. These results indicate the possibility that activation of angiogenic activity after resection of osteosarcoma tumors enhances progression of pulmonary metastasis. The current data also suggest that administration of antiangiogenic reagents can prevent progression of pulmonary metastasis in osteosarcoma postoperatively. PMID- 12567144 TI - Epithelioid hemangiomas of bone: spontaneous clinical and radiographic remission. AB - A case of pathologic multifocal epithelioid hemangioma which occurred in the perigestational period is reported. This is the only reported case in which an epithelioid hemangioma, without treatment, went into radiographic and clinical remission. This case may suggest an alternative treatment for epithelioid hemangioma. Rather than radiation therapy or amputation, close observation of this benign neoplasm may be indicated. PMID- 12567145 TI - Displaced acetabular fractures managed operatively: indicators of outcome. AB - This study examined retrospectively the clinical and radiographic features of acute displaced acetabular fractures which heavily influence the quality of an open reduction and clinical outcome. Of 424 acute fractures (119 simple and 305 associated injuries) in 411 patients, the mean time from the injury to surgery was 8.2 days (range, 0-21 days). The mean age of the patients was 46.5 years (range, 13-89 years), and followup was an average of 9.3 years (range, 3-21 years). For the 424 hips, the reduction was anatomic in 282 (67%), imperfect in 90 (21%), poor in 39 (9%), and secondarily congruent in 13 both-column fractures (3%). The quality of the reductions markedly deteriorated with advancing age. Clinically, the Harris hip score was excellent in 179 (42%), good in 126 (30%), fair in 54 (13%), and poor in 65 (15%). Excellent or good clinical outcomes were recorded in 249 of 282 patients (89%) with an anatomic reduction. Of the 119 patients with fair and poor results, 92 patients (77%) had complicating factors recognizable at clinical presentation, including extensive impaction, articular abrasion, a femoral head or neck fracture, or endogenous obesity. Although stable anatomic reduction of most displaced acetabular fractures affords the optimal prognosis, especially in younger patients, the initial clinical and radiologic evaluations delineate a sizable minority with poorly prognostic features that favor a poor clinical outcome after internal fixation, by minimizing the likelihood for an anatomic reduction or compromising the outcome despite an anatomic reduction. With the rapidly aging population accentuating this trend, a reappraisal for the role of the current therapeutic alternatives of nonoperative treatment, a limited open reduction, or an acute total hip replacement merits review. PMID- 12567146 TI - Operative management of lower extremity fractures in patients with head injuries. AB - Treatment of patients with lower extremity fractures and concomitant head injury is controversial. The authors compared reamed intramedullary nailing versus plating of femoral and tibial fractures in patients with polytrauma and concomitant head injury. One thousand five hundred twenty-five patients with head injuries were identified from a prospective trauma database. Of those, 1211 patients sustained severe head injuries (Abbreviated Injury Score >/= 3). One hundred nineteen patients with severe head injuries and lower extremity long bone fractures met the inclusion criteria. Ultimately, four patient groups were identified: Group A, reamed femoral nail (n = 21); Group B, femoral plate (n = 29); Group C, reamed tibial nail (n = 23); and Group D, tibial plate (n = 46). Reamed intramedullary nails did not significantly alter the risk of mortality when compared with plates in femoral (relative risk 0.46; 95% confidence interval, 0.04-4.6) and tibial (relative risk 1.18; 95% confidence interval, 0.05 11.9) fractures. The severity of the initial head injury (Glasgow Coma Scale score) was the strongest predictor of mortality. Functional independence scores between patients with reamed nails and patients with plates were similar at 1 year. Head injury does not seem to be a contraindication to reamed intramedullary nailing in patients with lower extremity fractures. The severity of head injury alone is an important predictor of outcome. A large, randomized trial with sufficient study power is needed to clarify this issue. PMID- 12567147 TI - Gluteus medius tendon injury during reaming for gamma nail insertion. AB - The use of intramedullary hip screws such as the Gamma nail has become a popular option for treatment of proximal femur fractures. The manufacturer recommends that the Gamma nail be inserted at the junction of the anterior (1/3) and posterior (2/3) of the tip of the greater trochanter and on the tip in the same region that the gluteus medius inserts. The purpose of the current cadaveric study was to quantify the damage to the gluteus medius tendon secondary to reaming of the greater trochanter necessary for safe Gamma nail insertion. The gluteus medius was identified and its tendinous insertion was measured. A Kirschner wire was placed in the recommended entry point using the assistance of C-arm fluoroscopy and a 17-mm reamer was advanced over the wire. The tendon then was inspected to evaluate the amount of intrasubstance damage. In 27 of 34 specimens, the portal was contained completely within the tendinous insertion of the gluteus medius. The percentage of the tendon insertion disrupted by the reamer ranged from 14.8% to 52.5% with an average of 27%. During appropriate placement of the Gamma nail, damage to the gluteus medius tendon is unavoidable and should be recognized as a potential cause of postoperative morbidity. PMID- 12567148 TI - Conehead wedging screw for distal radius fractures in elderly patients. AB - A retrospective study was done of the treatment of 76 distal radius fractures in patients older than 55 years, using intrafocal fixation with a conehead wedging screw or a threaded Kirschner wire. The average age of the 57 women and 19 men was 65.9 years. The fracture types classified according to Frykman were: 10 Type I, four Type II, two Type III, 11 Type IV, eight Type V, 10 Type VI, eight Type VII, and 23 Type VIII. The fixation materials used were a threaded Kirschner wire in 37 patients, and a conehead wedging screw in 39 patients. Followups for the two groups were 28 and 26 months, respectively. The conehead wedging screws were superior to the threaded Kirschner wires in maintaining postoperative reduction in displaced, moderately comminuted intraarticular fractures in elderly patients and had a lower complication rate. Intrafocal fixation using a conehead wedging screw may be indicated for treatment of intraarticular fractures with moderate comminution in elderly patients. PMID- 12567149 TI - Arthroscopic treatment of septic arthritis of the hip. AB - Arthroscopic debridement and drainage was done in 10 patients with septic arthritis of the hip. Staphylococcus aureus was the most commonly identified organism. All patients were placed in the supine position on a fracture table and three arthroscopic portals were used during the procedure. No major complication related to the arthroscopy of the hip resulted. The average followup was 4 years 11 months. All patients had excellent results. PMID- 12567150 TI - Early events in cartilage repair after subchondral bone microfracture. AB - The current study investigated healing of large full-thickness articular cartilage defects during the first 8 weeks with and without penetration of the subchondral bone using microfracture in an established equine model of cartilage healing. Chondral defects in the weightbearing portion of the medial femoral condyle were made bilaterally; one defect in each horse was microfractured whereas the contralateral leg served as the control. The expression of cartilage extracellular matrix components (Types I and II collagen and aggrecan) was evaluated using histologic techniques, reverse transcription coupled polymerase chain reaction, in situ hybridization, and immunohistochemistry. This study confirms an increase in Type II collagen mRNA expression in repair tissue as early as 6 weeks after microfracture. Although other matrix mRNA and protein levels changed in concentration and tissue location over the course of the study, no significant differences were seen in microfractured defects. Although the microfracture techniques appear to improve clinical functionality, volume of repair tissue, and augment Type II collagen content, aggrecan content is less than ideal. Therefore, methods to enhance key matrix components such as aggrecan after microfracture may additionally improve repair tissue observed after the procedure. PMID- 12567151 TI - Magnetic resonance discography in cadavers: tears of the annulus fibrosus. AB - The purpose of this study was to investigate the diagnostic capabilities of magnetic resonance discography in the detection of tears of the annulus fibrosus, and to compare the findings with those derived from conventional magnetic resonance imaging. Twenty-four intervertebral discs from cadavers were examined with magnetic resonance imaging and magnetic resonance discography, and anatomic correlation was done. Forty-six annular tears were documented on anatomic inspection of the specimens. Magnetic resonance discography allowed significantly better observation of radial than transverse annular tears, and identification of radial tears was significantly better than that of concentric tears. With magnetic resonance discography, detectability of concentric tears was not significantly better than that of the transverse tears. No significant differences in the detection of different types of annular tears were appreciated on the magnetic resonance images obtained before discography. Magnetic resonance discography allowed direct diagnosis of radial, transverse, and concentric tears in 100%, 57%, and 21% of specimens, respectively. With magnetic resonance imaging, the diagnosis of radial, transverse, and concentric tears was possible in 67%, 71%, and 21% of specimens, respectively. Magnetic resonance discography does well in the evaluation of the clinically important radial tears of the annulus fibrosus, although not uniformly well in the identification of other types of annular tears. PMID- 12567152 TI - Effect of pelvic tilt on acetabular retroversion: a study of pelves from cadavers. AB - Pelvic inclination is difficult to control on a standard radiograph of the pelvis and has a direct influence on the appearance of acetabular version. By defining the normal range of the distance between the symphysis and the sacrococcygeal joint on 86 standard anteroposterior radiographs of pelves a technique was developed to evaluate pelvic inclination. A statistically significant correlation between this distance and pelvic inclination was shown in four cadaver pelves. Acetabular retroversion signs (cross-over, posterior wall signs) were evaluated on normal pelves from cadavers (two females, two males) after mounting on a holding device and wire marking of the acetabular rims. Radiographs were taken 3 degrees stepwise through the range of 9 degrees inclination to 12 degrees reclination. At the neutral position, two acetabula appeared with both positive retroversion signs compared with none at 6 degrees reclination. At 9 degrees pelvic inclination all acetabula had both signs. Retroversion signs were significantly more pronounced and found at lower pelvic tilt angles in the pelves from males than from females. Evaluation of pelvic inclination will help to decrease measurement errors in assessment of acetabular orientation and femoral head coverage. This will be valuable in preoperative planning of reorientation procedures. PMID- 12567154 TI - Trabecular bone strain changes resulting from partial and complete meniscectomy. AB - Previous studies have documented how partial and complete meniscectomy affect articular contact pressure, but changes in load transfer through the complete osteochondral structure of the proximal tibia after partial and complete meniscectomy are not well known. The current study measured trabecular bone strain changes in the medial tibial plateau resulting from partial and complete medial meniscectomy. Midcoronal sections were prepared from knees from cadavers. High quality digital images, made from contact radiographs of loaded samples, were compared with digital images of unloaded samples using in-house software to measure trabecular bone strain. Measurements were made on specimens with an intact medial meniscus, after removal of the inner (2/3) of the meniscus, and after complete meniscectomy. Partial meniscectomy caused minimal increases in trabecular bone strain throughout the proximal tibia. However specimens with complete meniscectomy had significant trabecular bone strain increases. Many patients sustaining meniscus tears are young, therefore, it is important to understand mechanical changes associated with partial meniscectomy. The data suggest partial meniscectomy causes little change in load transfer through the proximal tibia, supporting partial meniscectomy as a good surgical option for patients with meniscus tears. PMID- 12567155 TI - Calf mass in an 11-year-old girl. PMID- 12567153 TI - Messenger ribonucleic acid levels in disrupted human anterior cruciate ligaments. AB - Thirty patients had anterior cruciate ligament reconstruction for ongoing instability. Two groups were defined according to gross morphologic features identified during reconstruction: anterior cruciate ligament disruptions with scars attached to a structure in the joint and disruptions without reattachments. Reverse transcription polymerase chain reaction for a subset of extracellular matrix molecules, proteinases, and proteinase inhibitors was done on samples of scarred anterior cruciate ligament tissue removed during reconstructive surgery. Results of the nonattached scar group showed significantly increased mRNA levels for Type I collagen, and an increased Type I to Type III collagen ratio compared with that for the attached scar group. In the first year after injury, decorin mRNA levels in the nonattached scar group also were significantly higher than in the attached scar group. Biglycan mRNA levels in the nonattached scar group correlated closely with Type I collagen mRNA levels. These results suggest differences in cellular expression in torn anterior cruciate ligaments that attach to structures in the joint versus those which do not. Although the molecular mechanisms responsible for these differences have not been delineated, different molecular signals may influence the gross morphologic features of anterior cruciate ligament disruptions or alternatively, differing gross morphologic features may be subject to different mechanical loads leading to altered molecular expression. However, the finding of endogenous cellular activity in injured anterior cruciate ligaments raises the possibility that this activity may be enhanced to improve outcomes. PMID- 12567156 TI - Topiramate for the treatment of kleptomania: a case series and review of the literature. AB - Kleptomania--the inability to resist the impulse to steal objects, not for personal use or monetary gain--is currently classified in psychiatric nomenclature as an impulse control disorder. There is no standard pharmacologic therapy for this disorder. If kleptomania was considered a form of obsessive compulsive disorder, treatments used for this spectrum, including serotonin reuptake inhibitors (SSRI), other antidepressants, opioid receptor antagonist medications, and mood stabilizers, could be logically tested. Topiramate is currently used for the treatment of patients with affective and compulsive eating disorders. This report documents three kleptomanic patients who responded well to topiramate given either alone or in combination with SSRIs. PMID- 12567157 TI - Acute delirium, delusion, and depression during IFN-beta-1a therapy for multiple sclerosis: a case report. AB - Adverse effects of interferon (IFN) treatment are common, and efforts to minimize these reactions are of considerable importance. IFN-beta-1a is an established therapy for patients with relapsing-remitting multiple sclerosis (MS). Its psychiatric side effects are debated and not yet fully established. The authors report here the case of a patient on IFN-beta-1a therapy for MS who developed acute delirium, delusion, and depression that ceased with treatment discontinuation. Although he had a history of recurrent major depressive disorder, his prior psychiatric illness had followed a course that was clinically independent of other signs of MS. This observation points out psychiatric vulnerability of patients taking IFN-beta-1a therapy for MS and suggests that IFN beta-1a may induce or exacerbate preexisting psychotic symptoms. PMID- 12567158 TI - Clozapine in Parkinson's disease psychosis: 5-year follow-up review. AB - The objective of this study was to monitor the long-term effect of clozapine administered to Parkinson's disease (PD) patients with psychosis. Confusion, visual hallucinations, and psychosis are major dose-limiting factors for long term dopaminergic management of PD. Classic neuroleptic agents exacerbate the motor symptoms of the disease. For this reason, the introduction of atypical antipsychotic drugs has been a major advancement for the management of psychosis in patients with PD. Of them, clozapine is one of the most effective. Thirty-two patients (mean age, 73 years; mean disease duration, 12.2 years) with PD and psychosis (DSM-IV), 14 of them with dementia (DSM-IV), were followed for 5 years with periodic clinical evaluation, Mini Mental State Examination (MMSE), and Parkinsonian Psychosis Rating Scale (PPRS) administered before and following the study (at least once in 6 months). Periodic blood count was performed for tracking neutropenia. Nineteen patients (8 with dementia) have continued to receive clozapine (mean daily dose, 50 mg). Thirteen patients stopped medication: 9 because symptoms improved and did not return after weaning off clozapine; 3 patients because of somnolence; and 1 because of personal reasons. The average duration of treatment in those in whom medication was stopped was 8.5 months (range, 1-24 months). No correlation was found between age, sex, duration, and severity of disease (Yahr scoring), the presence of dementia, and the response to clozapine. Also, the PPRS scoring did not influence clozapine response. No case of neutropenia was found. According to the experience accumulated and the results of the present study, the authors believe clozapine is the best therapeutic choice currently available for the management of psychosis in patients with PD. PMID- 12567159 TI - Beneficial effect of donepezil augmentation for the management of comorbid schizophrenia and dementia. AB - Comorbid schizophrenia and dementia is a common clinical phenomenon; however, management of the coexisting illnesses remains incomplete. Donepezil, a cholinesterase inhibitor, may be beneficial for the management of symptoms of Alzheimer's disease, a disease in which cholinergic pathways in the cerebral cortex and basal forebrain are well known to be compromised. Furthermore, impaired cognition in elderly schizophrenic patients has been observed to be more than two thirds; however, there are no published controlled studies reporting the use of cholinesterase inhibitors in the management of schizophrenia in patients with associated dementia. In this study, six patients with chronic schizophrenia and comorbid dementia were administered donepezil, 5 mg, in single-blind fashion as augmentation to their standard antipsychotic medication for a 4-week period. Patients were evaluated with the Mini Mental State Examination (MMSE); Alzheimer's Disease Assessment Scale, Cognitive subscale; Positive and Negative Symptom Scale (PANSS); and the Clinical Global Impression (CGI) scales. A significant improvement was noted in MMSE scores (P < 0.01) and for CGI scores (P < 0.01). In addition, three patients demonstrated improvement on the PANSS. Donepezil appears to be an effective treatment for the management of symptoms of dementia accompanying patients with comorbid schizophrenia and dementia. Since cholinergic dysfunction may be present in some patients with schizophrenia, the authors' findings further demonstrate the possibility that this disorder may be managed with cholinergic medications as augmenting agents, at least in this specific subpopulation of patients with comorbid dementia. To confirm the findings of this preliminary trial, further investigation is mandated with a larger sample of subjects in the context of a double-blind medication trial. PMID- 12567160 TI - Minocycline and other tetracycline derivatives: a neuroprotective strategy in Parkinson's disease and Huntington's disease. PMID- 12567161 TI - Effect of late initiation of levodopa treatment in patients with long-standing Parkinson's disease. AB - The time of initiation of levodopa therapy in patients with Parkinson's disease (PD) is still debatable, as is the hypothesis of levodopa toxicity Some researchers argue that late initiation of treatment will delay the appearance of response fluctuations. In the present study, 11 patients in whom treatment with low doses of levodopa was delayed for a mean of 7.9 +/- 3.1 years were followed for a mean of 15.7 +/- 3.3 years. Time of onset of response fluctuations and disease severity were compared with those in 17 patients with fluctuating PD who were treated with levodopa from disease onset. There was no significant change in time to onset of response fluctuations and dyskinesias once levodopa treatment was started, and late initiation of levodopa did not affect disease progression. The authors conclude that the decision of when to initiate levodopa treatment should be taken according to the patient's needs. PMID- 12567163 TI - Clinical pharmacology of antiepileptic drugs. PMID- 12567162 TI - Treatment of multiple sclerosis and related disorders: what's new in the past 2 years? PMID- 12567164 TI - The clinical and economic consequences of screening young men for genital chlamydial infection. AB - BACKGROUND: Wide-scale application of urine-based screening of asymptomatic men for chlamydial infection has not been thoroughly assessed. GOAL: The goal was to compare clinical and economic consequences of three strategies: (1). no screening, (2). screening with ligase chain reaction (LCR) assay of urine, and (3). prescreening urine with a leukocyte esterase test (LE) and confirming positives with LCR. STUDY DESIGN: We used a decision analytic model. RESULTS: At a chlamydia prevalence of 5%, the no screening cost was US dollars 7.44 per man screened, resulting in 522 cases of pelvic inflammatory disease (PID) per 100000 men. LE-LCR was most cost-effective, preventing 242 cases of PID over no screening at an additional cost of US dollars 29.14 per male screened. The LCR strategy prevented 104 more cases of PID than LE-LCR but cost US dollars 22.62 more per male screened. For this to be more efficient than LE-LCR, the LCR assay cost needed to decline to or = 86%. Nine phena were identified as Vibrio alginolyticus, V. fischeri, V. harveyi, V. carchariae, V. mediterranei, V. splendidus, V. furnissii, V. parahaemolyticus, and Photobacterium damselae subsp. damselae. The two latter comprised strains isolated from diseased fish. PMID- 12567242 TI - Seasonal population changes in the restriction fragment length polymorphism (RFLP) patterns from PCR-amplified 16S rRNA genes of predominant ribotypes in microbial mat samples from the Ebro Delta (Spain). AB - The stratified benthic microbial communities, which developed as a consequence of the physico-chemical gradients and the physiology of the inhabiting microorganisms, from Ebro Delta microbial mats were analyzed. 16S rRNA eubacterial and archaeal genes were amplified by polymerase chain reaction (PCR). PCR products were separately digested with three different restriction enzymes (AluI, HinfI, and RsaI) and later separated by horizontal electrophoresis to determine whether changes of predominant ribotypes are occurring over a period of a year. Comparison of total restriction patterns was performed by scoring similarities by using the Jaccard coefficient and then building a multidimensional scaling (MDS) map from the resulting similarities matrix. The three enzymes gave a consistent result, a seasonal distribution instead of a spatial and/or physiological one. PMID- 12567243 TI - Isolation and characterization of cyanobacteria from microbial mats of the Ebro Delta, Spain. AB - Isolation and identification of several strains of cyanobacteria from microbial mats of the Ebro Delta, Spain, are described. A series of tenfold dilutions was the first step of isolation. Self-isolation techniques, which use one or several physiological characteristics of a cyanobacterium, were applied in some cases to obtain enrichment cultures. Twelve filamentous strains were isolated and stored in axenic culture. As only a few cyanobacterial species can be frozen and revived without any cryoprotective agent, preservation of isolated strains was assayed with several cryoprotective solutions. Methanol and glycerol were not suitable as cryoprotective agents for most of the isolates. Dimethyl sulfoxide (DMSO) was apparently the best cryoprotector. A new method, which used a filter paper as a growing substratum that later could be directly stored at -80 degrees C, was successfully used. A morphological study of each strain under light and electron microscopy was made to classify them. All isolated strains belong to phylum BX, Class 1, subsection III of the Bergey's manual of systematic bacteriology, 2nd ed., vol. 1. Most genera are included in the LPP group as Lyngbya aestuarii and Microcoleus chthonoplastes. PMID- 12567244 TI - Regulation of extracellular chitinases and proteases in the entomopathogen and acaricide Metarhizium anisopliae. AB - Metarhizium anisopliae infects insects and ticks via a combination of specialized structures and cuticle degradation. Hydrolytic enzymes are accepted as key factors for the penetration step. The search for pathogenicity determinants has demonstrated that the process is multifactorial. Host specificity is an important factor to be addressed. The study of the enzymes produced during infection is important to discover those with a role in the process. To address some of the enzymes that take part during the infection of the tick, Boophilus microplus, we have analyzed the secretion of proteases and chitinases in single and combined carbon/nitrogen sources as compared with such complex substrates as chitin and B. microplus cuticles. Two chitinases, endo- and N-acetylglucosaminidases, and two proteases, subtilisin and trypsin-like proteases, were analyzed. Enzyme activities were detected in all carbon sources tested, but higher levels were found when combinations of carbon sources were used. A major 30-kDa protein apparently secreted during M. anisopliae growth on all carbon/nitrogen sources tested was demonstrated by SDS-PAGE. PMID- 12567245 TI - Replacement of methionine 208 in a truncated Bacillus sp. TS-23 alpha-amylase with oxidation-resistant leucine enhances its resistance to hydrogen peroxide. AB - The methionine residues at positions 17, 104, 208, 214, 292, 315, 324, and 446 in the primary amino acid sequence of a truncated Bacillus sp. TS-23 alpha-amylase (His(6)-tagged BLADeltaNC) was changed to oxidative-resistant leucine by site directed mutagenesis. The mutant enzymes with an apparent molecular mass of approximately 54 kDa were overexpressed in recombinant Escherichia coli. The specific activity for Met315Leu and Met446Leu was decreased by more than 76%, while Met17Leu, Met104Leu, Met208Leu, Met214Leu, Met292Leu, and Met324Leu showed 247, 128, 37, 260, 232, and 241%, respectively, higher activity than the wild type enzyme. In comparison with wild-type enzyme, a lower K(m) value was observed for all mutant enzymes. The 3.2- and 4.5-fold increases in the catalytic efficiency (k(cat)/K(m)) for Met208Leu and Met324Leu, respectively, were partly contributed by a 68% and 38% decrease in K(m) values. Wild-type enzyme was sensitive to chemical oxidation, but Met208Leu was stable even in the presence of 500 mM H(2)O(2). Except for Met214Leu, which was quite sensitive to H(2)O(2), the other mutants showed a profile of oxidative inactivation similar to that of the wild-type enzyme. These observations indicate that the oxidative stability of His(6)-tagged BLADeltaNC can be improved by replacement of the critical methionine residue with leucine. PMID- 12567246 TI - A Role for the glycolipid exoantigen (GLXA) in chlamydial infectivity. AB - The chlamydial glycolipid exoantigen, GLXA, is associated with the bacterial membrane, intracellular inclusion, and can also be found secreted into the microenvironment of Chlamydia trachomatis-infected cells. The aim of this study was to investigate the function of GLXA in chlamydial pathogenesis. Pretreatment of HeLa 229 cells with affinity-purified GLXA resulted in a significant enhancement of chlamydial infectivity as determined by inclusion body enumeration. The GLXA-mediated enhancement was shown to be time- and dose dependent and, more importantly, GLXA-specific, as the effect was abrogated by anti-GLXA antibody. In vitro neutralization assays on HEp-2 cells revealed that an anti-anti-idiotypic antibody to GLXA effectively reduced the infectivity of C. trachomatis, C. pneumoniae, and C. psittaci. In vivo, the co-inoculation of GLXA at the time of C. trachomatis serovar K intravaginal challenge of C3H/HeJ mice resulted in a significant increase in the numbers of shed organisms on days 4, 7, 14, 21, and 28. Taken together, these observations suggest that GLXA, both organism bound and secreted, is important in facilitating the initiation of infection. PMID- 12567247 TI - Effects of lysogeny of Shiga toxin 2-encoding bacteriophages on pulsed-field gel electrophoresis fragment pattern of Escherichia coli K-12. AB - Escherichia coli K-12 lysogens of three different Shiga toxin 2 (Stx2)-encoding bacteriophages were examined for variability in their pulsed-field gel electrophoresis (PFGE) fragment patterns. The PFGE fragment patterns could be classified into three types (i.e., PFGE types B, C, and D). For the PFGE type D, a 255-kbp fragment present in the original K-12 strain was apparently shifted by the size of Stx 2-encoding phage genomic DNA (ca. 65 kbp) to the position at 320 kbp. In contrast, the types B and C showed the above fragment shift plus further 6- and 10-fragment differences, respectively, from the original K-12 strain. The evidence suggests that even a single genetic event like lysogeny can cause marked genotypic modification of the host strain. PMID- 12567248 TI - Isolation, characterization, and antifungal susceptibility of melanin-deficient mutants of Scedosporium prolificans. AB - Scedosporium prolificans mutants lacking the ability to synthesize melanin were selected after ultraviolet light (UV) irradiation. UV exposure of S. prolificans conidia resulted in a high frequency of melanin-deficient (mel-) mutants. Stable and non-stable morphological variants were found in the population: reversion of the mutant phenotype was always to the wild-type phenotype. Based on their morphological differences, these variants were classified into five different groups that were phenotypically characterized. The mel- mutants plus the wild type strain were examined for in vitro susceptibility to antifungal agents with different and/or the same mechanism of action. There was no apparent difference in minimum inhibitory concentrations when comparing the wild-type and the mel- mutants. Therefore, melanin does not appear to confer protection against the more important antifungal agents in S. prolificans. PMID- 12567249 TI - A thermodynamic interpretation of the Monod equation. AB - The Monod equation for microbial growth is purely empirical, and the theoretical basis of this model stays unclear. Similar to any chemical reactions, overall microbial growth process is dependent upon the changes in free energy. This study showed that the Monod equation could be interpreted in a thermodynamic sense very well. It was probably for the first time demonstrated that the Monod constant ( K(s)) was inversely related to the equilibrium constant of the overall microbial growth process. PMID- 12567250 TI - Highly efficient paramagnetic labelling of embryonic and neuronal stem cells. AB - Recent developments in stem cell and gene therapy will require methods to monitor stem cell survival and integration repeatedly and non-invasively with a high temporal and spatial resolution in vivo. The aim of this study was to visualise embryonic and neuronal stem cells with standard contrast agents using a conventional clinical 1.5-Tesla scanner. We therefore modified standard transfection protocols including lipofection (Lipofectin and Lipofectamine) and calcium phosphate transfection for the efficient uptake of paramagnetic particles [gadolinium-diethylene triamine penta-acetic acid (Gd-DTPA)] in stem cells. Using this approach we obtained intracellular labelling efficiencies of up to 83%. Neither the proliferation capacity nor the differentiation efficiency was affected. Identical differentiation of labelled and unlabelled embryonic and neuronal cells was observed. The established labelling techniques used in this study displayed high labelling efficiencies in embryonic and neuronal stem cells without any alterations of cellular biology; therefore this approach might be a suitable method for targeting stem cells. PMID- 12567252 TI - [Pediatric dermatotherapy]. AB - Dermatological therapy of children differs considerably from treatment of adults and poses a special challenge. Dermatological diseases often form idiosyncratic entities with a prognosis different from adults and must be treated appropriately. Consideration should also be given to the fact that children occasionally experience adverse reactions to drugs, which are not observed in adults. A selection of dermatological diseases and typical drug reactions are presented to illustrate the special features and differences between pediatric dermatotherapy and treatment of adults. PMID- 12567253 TI - [Blistering autoimmune diseases of childhood]. AB - Autoimmune blistering skin diseases are exceedingly rare in children. In most cases, they are characterized by circulating autoantibodies directed against adhesion structures of the skin which subsequently induce blistering at the dermoepidermal junction or intraepidermally. The most frequent paediatric immunobullous disease is linear IgA dermatosis; all others such as pemphigoid, epidermolysis bullosa acquisita, dermatitis herpetiformis and pemphigus are less common than in adults. In children, mucous membranes are involved more frequently. Recent advances in the identification of the autoantigens have improved diagnostic approach and therapeutic management of blistering diseases. In most cases, treatment requires systemic immunosuppression. With respect to the chronic course of the diseases and potential complications of treatment, cooperation between dermatologists and paediatricians seems advisable. PMID- 12567254 TI - [Photodermatoses during childhood]. AB - Actinic dermatitis and phototoxic and idiopathic photodermatoses occur relatively often during childhood. Photodermatoses stemming from enzyme defects and genetic disorders are much less common, while photoallergic and chronic actinic dermatitis have not been dealt with to date. A specific sensitivity of a child's skin to light is often the first manifest clinical symptom of a photodermatosis, the key is to ensure early diagnosis. However, it is often difficult to reach a diagnosis, as the lesions are barely perceptible or have even healed by the time the patient is examined. This is why photoprovocation tests are so important in diagnosing photodermatoses, particularly during childhood. In addition to early diagnosis of these illnesses and offering the appropriate treatment, it is also crucial that the parents be well informed and aware of what symptoms to look for so that they can protect their children from overexposure to UV light and thereby avoid later actinic injuries that can become as serious as malignancies. PMID- 12567255 TI - [Xeroderma pigmentosum and related syndromes]. AB - Xeroderma pigmentosum, trichothiodystrophy and Cockayne-syndrome are rare, autosomal recessive genodermatoses, which are clinically heterogeneous. Generally, the first signs and symptoms appear at an early age. Although all three syndromes show photosensitivity and an underlying defect in the repair of UV-induced DNA damage, only patients with xeroderma pigmentosum have an increased skin cancer risk. The fact that all three of these syndromes can be caused by mutations in the same gene further emphasizes the role of these syndromes as an important model system for the pathogenesis of skin tumors. Recent findings in the fields of DNA repair, regulation of transcription and immunology have not only further unraveled the underlying mechanisms of these diseases but also provided important insights into the pathogenesis of skin tumors. The clinical characteristics of xeroderma pigmentosum and related DNA repair deficiencies are reviewed with an attempt to point out direct connections between clinical signs and symptoms and their underlying mechanisms. PMID- 12567256 TI - [Nail changes during childhood]. AB - Nail changes occur frequently in adults, but are relatively rare in children. There are, however, certain nail changes that emerge particularly during childhood. Ingrown nails, onychoschizia, congenital slanted alignment of the large toenail, and psoriasiform nail dystrophies can be frequently observed in toddlers and in schoolchildren the syndrome of an ingrown nail, nail artifacts, periungual warts or fibromas, trachyonychia, and nail mycoses. Connatal nail changes can appear as isolated events or together with other symptoms whereas acquired nail changes are caused by infections, traumatic injuries, or tumors, in conjunction with skin diseases or general illnesses, or even develop spontaneously. PMID- 12567257 TI - [Significance of immunologic tolerance in dermatology]. AB - The immunological tolerance processes enable the organism to distinguish between self and non-self and are, therefore, critical for an efficient immune system. Exogenous or endogenous factors that disturb tolerance mechanisms induce uncontrolled activation of the immune system and the development of autoimmune diseases. In the field of dermatology, the most relevant autoimmune diseases are connective tissue diseases and autoimmune bullous skin disorders. In contrast, increased activity of tolerance shuts down parts of the normal immune response and thus facilitates the development of neoplasia and microbial infections in the skin and other organs. Immunological mechanisms for the induction of tolerance have been studied with the help of experimental models of tolerance to contact allergens. T- and B-cells, as well as antigen presenting cells, in particular dendritic cells, are involved in the immunological mechanisms of tolerance. The modification of autologous immune cells of patients with malignant tumors, allergic and autoimmune diseases might have potential for the development of new therapies. PMID- 12567258 TI - [Drug interactions in clinical practice. A pilot project for quality assurance in prescribing]. AB - BACKGROUND AND OBJECTIVE: The aim of the project was to document and evaluate the safety of combinations of drugs prescribed during hospitalization, producing a method of quality assurance assessment. PATIENTS/METHODS: The medications of 396 patients already taken at home as well as the drugs given in the hospital during an acute illness were analyzed. Potential drug interactions were detected using a computerized drug-interaction checking system i.e. Micromedex CD-ROM database DRUG-REAX. This program differentiates between contraindicated, major, moderate or minor interactions. RESULTS: No contraindicated combinations of medications were found but in 265 instances (66,9%) potentially relevant ( major or moderate) combinations could be detected. There was a correlation between potentially dangerous drug combinations and the number of coadministered drugs. Despite the detection of a large number of potentially troublesome combinations, no significant clinical problems were observed over at least three days of observation. CONCLUSIONS: The number of potentially relevant drug interactions represents a severe problem in drug prescribing. PMID- 12567259 TI - [Apomorphine hyperpigmentation]. AB - A 72 year old bedridden, disoriented man presented with a continuously increasing number of blue nodules on his abdomen and both thighs. In addition, he had a melanoma on his left forearm (SSM, Clark level III, Breslow 0.75 mm), which lead to the clinical diagnosis of melanoma metastases. Biopsy of one of the blue nodules showed CD68 positive histiocytic cells loaded with brownish pigment granules and a lymphocytic infiltrate within the deep dermis and upper subcutis. The pigment reacted histochemically similarly to melanin. Melanocytes were absent at these sites. Because of the unexplained clinical and histopathological picture, the patient's history was reassessed and it was learned that the patient had received subcutaneous infusions of apomorphine for the past 10 years for the treatment of Parkinson's disease. By oxidation, apomorphine may be converted into tetrahydroisoquinoline-melanin, which apparently is the cause for the accumulation of pigment within the deep dermis. PMID- 12567260 TI - [Sporotrichosis as the cause of a leg ulcer]. AB - A young man presented at Lugala Lutheran Hospital (Tanzania) with an ulcer on his lower leg which had developed over the past 9 weeks. Subcutaneous nodules and plaques were found all the way up to his groin; this observation prompted a strong suspicion that the patient had the lymphocutaneous form of sporotrichosis which had not been seen at this hospital before. The diagnosis was confirmed histopathologically. The patient was then treated with saturated solution of potassium iodide. PMID- 12567261 TI - [First descriptions in the "International atlas of rare skin diseases" of 1886]. AB - In the history of dermatology, we find an outstanding example of scientific collaboration of leading dermatologists leading to an international publication in the late 19th century. Published by Malcolm Morris, Paul Gerson Unna, Henry Camille Chrysostome Leloir and Louis Adolphus Duhring, the "International Atlas of Rare Skin Diseases" focused on rare and unique cases in dermatology. This concept, extraordinary for the time of publication, was found to be very successful. Numerous first descriptions are found in the "International Atlas", such as porokeratosis of Mibelli, acanthosis nigricans of Pollitzer and Janovsky and also psorospermosis of Darier. The focus on distinct and detailed descriptions to promote progress in the science of dermatology sets this effort apart from all previous atlases. Therefore, the "International Atlas was one of the first steps leading to the modern concepts of scientific publication in dermatology. PMID- 12567262 TI - [Pityriasis rosea. Reactivation of human Herpesvirus 6 and 7 infection?]. PMID- 12567263 TI - Evidence of clinical and genetic heterogeneity in autosomal dominant congenital cerulean cataracts. AB - Autosomal dominant cerulean cataracts (ADCC) have previously been mapped to two loci: one on chromosome 17q24 and the other on chromosome 22q11.2-q12.2, which includes the beta-B2 crystallin (CRYBB2) candidate gene. Using polymorphic markers in these regions (D17S802, D17S836, D17S1806 and CRYBB2, D22S258) for linkage analysis, we excluded these loci in a large Moroccan family presenting with an unusual form of ADCC with early onset of lens opacities and rapid evolution. This finding confirms the clinical and genetic heterogeneity of autosomal dominant congenital cerulean cataracts. PMID- 12567264 TI - A pooled case-control study of the apolipoprotein E (APOE) gene in age-related maculopathy. AB - Age-related maculopathy (ARM) is a multifactorial disorder known to have a substantial genetic component. The epsilon4 allele of the apolipoprotein E gene (APOE-4) has previously been reported to have a protective effect on ARM risk, while the APOE-2 allele may increase disease risk. This study combined four independent data sets (three US and one European) of Caucasian ARM patients and controls in order to obtain better statistical power to examine the role of APOE in ARM. APOE genotype and allele frequencies were compared for 617 ARM cases and 1260 controls, adjusting for age and sex differences between the two groups via multiple logistic regression. The protective effect of the APOE-4 allele on ARM risk was confirmed (age- and sex-adjusted odds ratio (OR) for APOE-4 carriers 0.54, 95% confidence interval (CI) 0.41-0.70, p < 0.0001). The effect of APOE-4 did not differ significantly between males and females and was observed consistently for both atrophic and neovascular ARM. Evidence for an increased risk of ARM due to the APOE-2 allele was found for men, but not for women (OR for men 1.54, 95% CI 0.97-2.45; OR for women 0.74, 95% CI 0.52-1.06, p = 0.01 for interaction of sex and APOE-2 carrier status). These data confirm that the APOE-4 allele, or an allele in linkage disequilibrium with it, reduces the risk of ARM. They also suggest that the effect of the APOE-2 allele may vary by gender, and that APOE-2 may confer an increased risk only to males. PMID- 12567265 TI - A novel mutation disrupting the cytoplasmic domain of CRB1 in a large consanguineous family of Palestinian origin affected with Leber congenital amaurosis. AB - Leber congenital amaurosis (LCA) is a genetically heterogeneous autosomal recessive condition responsible for congenital blindness or greatly impaired vision since birth. Eight LCA loci have been mapped, but only six out of eight genes have been hitherto identified. A genome-wide screen for homozygosity was conducted in a large consanguineous family originating from Palestine, for which no mutation was found in any of the six known LCA genes and that excluded the LCA3 and LCA5 loci. Evidence for homozygosity, however, was found in all affected patients of the family on chromosome 1q31, a region in which the human homologue of the Drosophila melanogaster crumbs gene (CRB1) has been mapped. Consequently, we proposed a hypothesis that the disease-causing mutation in this family might lie in an unexplored region of this LCA gene. As a matter of fact, while no mutation was found in any of the 11 CRB1 exons originally reported, we identified a 10-bp (del 4121-4130) deletion segregating with the disease in a later reported 12th exon lying in the 3' end of the gene. Interestingly, this deletion disrupts an amino acid sequence that was shown to be crucial for the function of the protein in the Drosophila counterpart (CRB). PMID- 12567266 TI - TGFBI gene transcript is transforming growth factor-beta1-responsive and cell density-dependent in a human corneal epithelial cell line. AB - PURPOSE: To determine the roles of transforming growth factor-beta( 1) (TGF beta(1)), epidermal growth factor (EGF), and cell density on the regulation of the TGFBI gene transcript in a human corneal epithelial cell line (HCE-T). METHODS: HCE-T cells (40-50% confluent), which possess the biochemical and morphological phenotype of human corneal epithelial cells, were treated with either TGF-beta(1) (20 ng/ml) or EGF (20 ng/ml) for 0, 24, 48, and 72 h. Total RNA was isolated at each time point from the treated cells and the placebo treated controls. The TGFBI mRNA transcript level was quantitated using Northern blot analysis. RESULTS: TGF-beta(1) upregulates TGFBI gene transcript in this human corneal epithelial cell line, reaching a peak of 2.5-fold of upregulation at 48 h after TGF-beta(1) treatment and a 1.5-fold at 72 h. In contrast, EGF treatment showed no effect on the TGFBI mRNA level. In addition, the TGFBI gene transcript appears to be cell density-dependent because the transcript level shows a trend of reduction as the cell density increases from 40-50% to full confluency. CONCLUSION: The TGFBI gene transcript in this human corneal epithelial cell line is upregulated by growth factor TGF-beta( 1), but is not affected by growth factor EGF. Furthermore, cell density appears to be an important regulatory mechanism in controlling the level of TGFBI gene expression in corneal epithelial cells. PMID- 12567268 TI - Fanconi anemia and primary cataracts: first case. AB - Fanconi anemia or pancytopenia is an autosomal recessive condition presenting with a combination of pancytopenia with a mean age of onset of about eight years, a tendency to leukemia, and congenital anomalies. Although ocular abnormalities have been described, cataracts have not been previously reported. We present a patient with proven Fanconi anemia and cataracts. PMID- 12567267 TI - Corneal decompensation in a boy with Kearns-Sayre syndrome. AB - This paper describes the clinical history of a young boy with Kearns-Sayre syndrome. The first presenting symptom of Kearns-Sayre syndrome in this boy was corneal edema with photophobia and tearing. PMID- 12567271 TI - Antioxidant activity of piperlactam S: prevention of copper-induced LDL peroxidation and amelioration of free radical-induced oxidative stress of endothelial cells. AB - The protective effects of piperlactam S, an alkaloid isolated from Piper kadsura (Choisy) Ohwi, on lipid peroxidation and free radical-mediated cell injuries were investigated. Piperlactam S (1 to 20 microM) concentration-dependently prevented the copper-catalyzed oxidative modification of human low-density lipoproteins (LDL) measured through (i) the lag period, (ii) the slope of the propagation phase, (iii) the total amount of conjugated dienes formed, and (iv) the electrophoretic mobility of LDL. Fe2+-induced oxidative modification of cell membrane was also significantly attenuated by piperlactam S as measured by thiobarbituric acid-reactive substances (TBARS). Furthermore, piperlactam S effectively minimized the loss of cell viability induced by Fenton's reagent (H2O2/FeSO4) in cultured endothelial cells and significantly reversed H2O2/FeSO4 induced impairment of endothelium-dependent relaxation to acetylcholine in rat aorta. Since the oxidative modification of LDL plays an important role in the genesis of atherosclerosis, piperlactam S may help to reduce the risk of atherosclerosis, not only by protecting LDL and membrane lipids from oxidative modification but also by reducing free radical-induced endothelial injury and/or dysfunction. PMID- 12567273 TI - Cancer chemopreventive in vitro activities of isoflavones isolated from Iris germanica. AB - Six known isoflavones were isolated from the rhizomes of Iris germanica, and were established by UV, MS and NMR techniques as irisolidone (1), irisolidone 7-O alpha-D-glucoside (1a), irigenin (2), irilone (3), iriflogenin (4), and iriskashmirianin (5). These compounds were examined for their cancer chemopreventive potential. They were shown to be potent inhibitors of cytochrome P450 1A activity with IC 50 values in the range 0.25-4.9 microM. The isoflavones 2, 3 and 5 displayed moderate activity as inducers of NAD(P)H:quinone reductase (QR) in cultured mouse Hepa 1c1c7 cells, with CD values (concentration required to double the specific activity of QR) of 3.5-16.7 microM, whereas weak activity was observed with compounds 4 and 5 in the radical (DPPH) scavenging bioassay (IC 50 values 89.6 and 120.3 microM, respectively). With respect to anti-tumor promoting potential based on anti-inflammatory mechanisms, none of the compounds demonstrated significant activity in the concentration range tested. PMID- 12567272 TI - Anti-inflammatory properties of piperlactam S: modulation of complement 5a induced chemotaxis and inflammatory cytokines production in macrophages. AB - Macrophages infiltrate tissues in response to chemoattractants including complement 5a (C5a). Infiltrating macrophages clear microorganisms but also can cause tissue damage. We hypothesized that prevention of macrophages from excessive recruitment into infected sites may underlie the anti-inflammatory effects of piperlactam S, an alkaloid isolated from Piper kadsura (Choisy) Ohwi. To test this hypothesis, chemotactic migration of RAW264.7 macrophages was induced by C5a and the effects of piperlactam S were studied. The results showed that piperlactam S (1-30 microM) concentration-dependently suppressed C5a-induced migration across a fibrinogen-coated barrier with an IC50 of 4.5 +/- 0.3 microM. At 30 microM, piperlactam S inhibited chemotaxis by more than 95 % and also decreased phagocytosis by 25 % without reducing macrophage viability and adherent capacity. Furthermore, piperlactam S treated cells adhered but failed to spread and elongate as in control cells. Finally, piperlactam S inhibited the C5a stimulated release of tumor necrosis factor-alpha and interleukin-1beta. We conclude that retardation of macrophage recruitment by interfering with the migration process and suppression of cytokines production might underlie the potential usefulness of piperlactam S as an anti-inflammatory agent. PMID- 12567274 TI - Cytotoxic activity and G2/M cell cycle arrest mediated by antofine, a phenanthroindolizidine alkaloid isolated from Cynanchum paniculatum. AB - Starting with an extract derived from the root of Cynanchum paniculatum Kitagawa (Asclepiadaceae) that was active in the process of inhibiting the growth of human cancer cells in culture, a phenanthroindolizidine alkaloid antofine was isolated and identified as an active principle (IC50 = 7.0 +/- 0.2 ng/ml for A549, human lung cancer cells; IC50 = 8.6 +/- 0.3 ng/ml for Col2, human colon cancer cells). Prompted by the high potency of cancer cell growth inhibition, additional action mechanism studies were performed with antofine. Utilizing cultured Col2 cells as a model, antofine induced arrest in the G2/M phase of the cell cycle after 48 h of incubation. With wash-out experiment, colony formation was also inhibited in a dose-dependent manner. These data suggest the potential of antofine to serve as a cancer chemotherapeutic agent by virtue of arresting the cell cycle. PMID- 12567275 TI - Crude extract of Salvia miltiorrhiza and salvianolic acid B enhance in vitro angiogenesis in murine SVR endothelial cell line. AB - Salvia miltiorrhiza (SM) has been used clinically in Asian countries to improve the microcirculation in the human body. Although a pure compound extracted from SM, salvianolic acid B (Sal B), has been reported to be effective against fibrosis and ischemia-reperfusion injury, possibly through its anti-lipid peroxidation action, the effect of SM on angiogenesis remains unclear. It is our interest to investigate the role of SM on the regulation of the angiogenic process. By using the SVR endothelial cell line as an in vitro system, the effects of Sal B on the gene expression of vascular endothelial growth factor (VEGF) and its receptors VEGF-R1, VEGF-R2 were evaluated by morphology, differentiation assay, reverse-transcribed polymerase chain reaction (RT-PCR) and Western blot analysis. The results showed that both the crude extract of SM and the pure compound Sal B had enhancing effects on cell growth and differentiation. The gene expression of matrix metalloproteinase-2 (MMP-2) was up-regulated after Sal B treatment for 2 h, while VEGF and VEGF-R2 gene expression were up-regulated 40 min after Sal B treatment. We conclude that the crude extract of SM and Sal B enhance angiogenic processes on SVR cells through up-regulation of VEGF and VEGF receptors genes. PMID- 12567276 TI - Protective effects of honokiol and magnolol on tertiary butyl hydroperoxide- or D galactosamine-induced toxicity in rat primary hepatocytes. AB - The aim of this study was to investigate the protective effect of honokiol and magnolol on hepatocyte injury induced by either tertiary butyl hydroperoxide (tBH)- or D-galactosamine (GalN). The cellular leakage of LDH and AST, and cell death by treatment with 1.5 mM tBH for 1 h, were significantly inhibited by treatment with honokiol (40 and 20 microM) or magnolol (40 microM). Treatment with honokiol or magnolol significantly inhibited lipid peroxidation in both cells and media, the generation of intracellular reactive oxygen species (ROIs), and intracellular glutathione (GSH) depletion induced by tBH. The cellular leakage of LDH and AST, and cell death, by 24-hour treatment with 30 mM GalN were significantly inhibited by treatment with honokiol (20, 5 and 1 microM) or magnolol (20, 5 and 1 microM). Treatment with honokiol (20, 5 and 1 microM) or magnolol (20 and 5 microM) significantly inhibited the intracellular GSH depletion induced by GalN. The hepatoprotective effects of honokiol and magnolol on oxidative stress induced by tBH were probably the result of their antioxidant activity. Honokiol and magnolol also had a protective effect against GalN-induced hepatotoxicity, which was used as an alternate model to oxidative stress, acting by inhibiting intracellular GSH depletion. PMID- 12567277 TI - Galphimine B modulates synaptic transmission on dopaminergic ventral tegmental area neurons. AB - Galphimine B (GB) is a bioactive compound isolated from the plant Galphimia glauca Cav. (Malpighiaceae) and has been shown to have central nervous system depressant properties. In an earlier study, it was reported that both systemic and local administration of GB modified the extracellular spontaneous spiking activity in the ventral tegmental area (VTA) neurons. In the present study we analyzed the synaptic effects of this compound on dopaminergic neurons. Recordings were made in brain slices using intracellular and patch-clamp techniques, in dopaminergic (DA) VTA neurons. Spontaneous miniature excitatory postsynaptic currents (mEPSCs), excitatory postsynaptic currents (EPSCs) and inhibitory postsynaptic potentials (IPSPs) were recorded in control situation and after bath infusions of several concentrations of GB (1 microM-5 mM), GABA (1 nM 100 microM) and the GABA A blockers, picrotoxin (100 microM) and bicuculline (10 microM), and GABA B blocker saclofen (200 microM). GB administration reduced the frequency (p < 0.05) but not the amplitude of mESPCs. However, GABA (IC 50 = 645 nM) and GB (IC 50 = 174.5 microM) infusion significantly reduced the amplitude of stimuli induced EPSCs, Bicuculline (10 microM) co-administration only reduced GABA effects and did not modify the GB depressant action. Finally, isolated GABAergic IPSPs were modified by the addition of picrotoxin, but GB had no effect on these evoked synaptic responses. The present results indicate that GB modifies synaptic activity on dopaminergic VTA neurons by a non-GABAergic mechanism. PMID- 12567278 TI - Physiological responses of a natural antioxidant flavonoid mixture, silymarin, in BALB/c mice: III. Silymarin inhibits T-lymphocyte function at low doses but stimulates inflammatory processes at high doses. AB - Silymarin is a mixture of bioactive flavonoids isolated from Milk Thistle (Silybum marianum). Crude extracts from this plant have been used for centuries as a natural remedy and silymarin is now effectively used in the treatment of inflammatory liver toxicity and disease in humans. In vitro studies show that silymarin can inhibit the production and damage caused by tumor necrosis factor alpha (TNFalpha) and is a potent antioxidant both in vitro and in vivo. Such findings suggest silymarin may impact the immune system but little information exists following in vivo exposure. Therefore, we tested the hypothesis that exposure to silymarin will modulate the inflammatory immune response. Male BABL/c mice (6/group) were treated intraperitoneally once daily for five days with 0, 10, 50 or 250 mg/kg of silymarin. Silymarin exposure did not produce any signs of overt toxicity or any changes in relative organ weights. Flow cytometric examination of splenic lymphocyte populations showed that the absolute number of CD3+ T-lymphocytes was reduced in the 10 and 50 mg/kg groups although significance was evident only in the 10 mg/kg group. Concomitant decreases in CD4+ and CD8+ T-cell populations were observed but only the CD4+ population in mice treated with 10 mg/kg of silymarin was significantly different from control. Functional examination of secondary lymphoid cells revealed that phytohemagglutinin-induced T-lymphocyte proliferation was increased in the lowest dose group only. B-lymphocyte blastogenesis induced by lipopolysaccharide was increased following exposure to 10 and 50 mg/kg of silymarin. Similarly, expression of TNFalpha, inducible nitric oxide synthase, IL-1beta and IL-6 mRNA were increased dose-dependently. The expression of IL-2 and IL-4 were reduced in mice treated with 10 and 50 mg/kg of silymarin although only the 10 mg/kg group was significantly different from control. The results indicate that in vivo parenteral exposure to silymarin results in suppression of T-lymphocyte function at low doses and stimulation of inflammatory processes at higher doses. Further studies investigating the effects of silymarin on the immune system are warranted. PMID- 12567279 TI - Acute drought stress and plant age effects on alkamide and phenolic acid content in purple coneflower roots. AB - The effects of acute periods of drought stress on dry weight, and alkamide and phenolic acid content in purple coneflower [Echinacea purpurea (L.) Moench, Asteraceae] roots are described. Plants subjected to brief drought stress periods for two seasons during the initial flowering stage (D-F2) produced fall-harvested roots with significantly greater cichoric acid concentration (mg/g) than corresponding well-watered controls of the same age (C-2). Total alkamide, including the tetraenoic acid isomers, and chlorogenic acid concentrations from fall-harvested roots were largely unaffected by drought stress, regardless of when the stress occurred developmentally. The alkamide concentration in three year roots was significantly less than that in two-year roots, with an average decrease of 50.5 %. Conversely, total phenolic acids increased an average of 67.1 % for all treatments from two to three years of age. Root dry weight increased significantly by an average of 70.0 % for all drought-stressed plants from two to three years of age, compared to an increase of 35.2 % for well-watered controls. The results suggest that controlled drought stress can stimulate increased root dry weight and root cichoric acid content, and that root age is the predominant factor determining overall phytochemical content variation. PMID- 12567280 TI - Isolation and characterization of anticomplementary beta-glucans from the shoots of bamboo Phyllostachys edulis. AB - Bamboo, Phyllostachys edulis produces well known edible shoots in Asia. Immunostimulating anticomplementary (complement activating) substances have been recognized as a characteristic biological response modifier (BRM). In the present study, we isolated and characterized three anticomplementary beta-glucans (BS BGA, BS-BGB, and BS-BGC) from bamboo shoots. Hot-water extraction, DEAE-Toyopearl 650M-column chromatography, amylase digestion and concanavalin A adsorption, and Sephacryl S-100 HR column chromatography were applied to isolate the beta glucans. The average molecular masses of the beta-glucans were estimated to be from 14,500 to 85,300 Da by HPSEC-MALLS-RI. All three beta-glucans (0.1-1.0 mg/mL) activated the complement system via the alternative pathway, and could cleave human complement C3 under Ca 2+-free gelatin veronal buffered saline. Among them, the largest molecule, BS-BGA was the most potent complement activator. Methylation analysis and NMR spectroscopy were used to achieve their structural characterization. They are all water-soluble and composed mainly of backbone structures of beta-(1-->3)-glucan with beta-(1-->4)-linked side chains varying in degree of branching. BS-BGA consisted of a higher proportion of 3 linked glucopyranosyl residues and a lower degree of branching than BS-BGB and BS BGC. In particular, BS-BGA contained a small amount of O-acetyl groups at C-6 of the 3-linked glucopyranosyl residues. These data demonstrate that the structural characteristics including molecular size, degree of branching, and O-acetyl substitution are involved, at least in part, in their different anticomplementary activities. PMID- 12567281 TI - Lignans with inhibitory activity against NFAT transcription from Schisandra chinensis. AB - Seven lignans from Schisandra chinensis were investigated for their inhibitory activity on NFAT transcription. Both gomisin N (IC 50 : 1.33 +/- 0.05 microM) and schisandrol A (IC 50 : 1.34 +/- 0.05 microM) showed higher activity than gomisin E (IC 50 : 4.73 +/- 0.09 microM), schisandrin A (IC 50 : 7.23 +/- 0.21 microM), schisandrin C (IC 50 : 7.54 +/- 0.22 microM), benzoylisogomisin O (IC 50 : 11.06 +/- 1.02 microM) and schisandrol B (IC 50 : 16.37 +/- 1.00 microM). PMID- 12567282 TI - Acteoside from Cistanche salsa inhibits apoptosis by 1-methyl-4-phenylpyridinium ion in cerebellar granule neurons. AB - In this study we assessed the effect of acteoside that significantly improved cell viability and inhibited lactate dehydrogenase (LDH) release. Furthermore acteoside prevented a neurotoxin 1-methyl-4-phenylpyridinium ion (MPP+)-induced apoptosis in CGNs. Accordingly, our flow cytometric analysis of CGNs after acteoside treatment revealed a decrease in the number of the MPP+-induced apoptotic cells (P < 0.001). Western blot analysis demonstrated that acteoside inhibits the active caspase-3 fragment (17 kDa) (P < 0.001) and the proteolytic poly (ADP-ribose) polymerase (PARP) fragment (85 kDa) expression (P < 0.001) following MPP + treatment in CGNs. We conclude that acteoside prevents the MPP+ induced apoptosis and inhibits the apoptosis-related pathway. PMID- 12567283 TI - Cytotoxic effect of the diterpene lactone dehydrocrotonin from Croton cajucara on human promyelocytic leukemia cells. AB - Diterpenes exhibit potent antineoplastic properties against human and murine carcinoma cell lines. trans-Dehydrocrotonin from Croton cajucara, a Brazilian medicinal plant, is a nor-diterpene with antiulcerogenic activity. In this work, we examined the effect of trans-dehydrocrotonin (t-DCTN) on the vitality of HL60 cells by assessing the MTT reduction, protein content and phosphatase activity of these cells. Protein quantification indicated that t-DCTN reduced the number of cells with an IC50 of 500 microM; mitochondrial function (MTT reduction), was also inhibited (IC50 = 300 microM), when the cells were treated for 24 h. In contrast, when the cells were treated with this lactone in the initial plating and cultured for 96 h, t-DCTN was more toxic for all parameters analyzed: MTT and phosphatase activity (IC50 = 180 microM) and protein content (IC50 = 150 microM). The flavonoid utilized as positive control myricetin and the following IC50 values were obtained after 24 h of treatment: 300 and 192 microM for protein content and MTT reduction, respectively. According to the chemical characteristics of both compounds, the cytotoxic effect of t-DCTN could be explained through two mechanisms: adduct formation with DNA and proteins and/or oxidative stress induction. PMID- 12567284 TI - Potential antitumor promoting diterpenoids from the stem bark of Thuja standishii. AB - Six diterpenes, including one new natural product, were isolated from a CHCl 3 extract of the stem bark of Thuja standishii. The new compound has been characterized as 15-oxolabda-8(17),13 Z-dien-19-oic acid. The known compounds were identified as ferruginol (2), sugiol (3), isocupressic acid (4), sandaracopimaric acid (5) and 15-oxolabda-8(17),13 E-dien-19-oic acid (6). Compounds 2-5 and the derivatives 4a and 4b were tested for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O tetradecanoylphorbol 13-acetate (TPA). Compounds 2, 3, 4 and 5 showed strong inhibitory effect on EBV-EA induction (100 % inhibition at 1000 mol ratio/TPA). PMID- 12567285 TI - Inhibition of 5alpha-reductase activity by diarylheptanoids from Alpinia officinarum. AB - The acetone extract from the rhizomes of Alpinia officinarum was evaluated for activity against 5alpha-reductase which had been prepared from rat prostate. The fraction responsible for the inhibition of the enzyme was purified, analyzed, and the active constituents were identified as four diarylheptanoids, 1,7 diphenylhept-4-en-3-one, dihydroyashabushiketol (1,7-diphenyl-5-hydroxy-3 heptanone), 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone and 5 hydroxy-7-(4"-hydroxyphenyl)-1-phenyl-3-heptanone. PMID- 12567286 TI - The vasorelaxant activity of marrubenol and marrubiin from Marrubium vulgare. AB - Crude extracts of the aerial parts of Marrubium vulgare show a potent in vitro inhibition of KCl-induced contraction of rat aorta. Bio-guided fractionations, spectroscopic analysis and chemical derivatization revealed the furanic labdane diterpenes marrubenol and marrubiin as the most active compounds. PMID- 12567287 TI - Essential oil composition and antimicrobial activity of Santiria trimera bark. AB - The composition and the antimicrobial activity of the bark oil of Santiria trimera (Oliv.) Aubrev., a plant widely used by the traditional healers in S. Tome and Principe, especially for wound healing, are reported for the first time. The analysis of the essential oil was carried out by GC and GC-MS. The oil contains a high content of monoterpenes, alpha-pinene (66.6 %) being the major constituent, followed by beta-pinene (20.0 %). The essential oil was active against both bacteria and fungi strains, except Staphylococcus epidermidis and Aspergillus niger. It exhibited significant antimicrobial activity against Proteus vulgaris and Cryptococcus neoformans with MICs values of 1.11 microl/ml and lower than 0.71 microl/ml, respectively. PMID- 12567288 TI - Antimicrobial activity and chemical composition of the essential oil of Lippia graveolens from Guatemala. AB - The composition and the antimicrobial activity of the essential oil of Lippia graveolens H.B.K. collected in Guatemala were studied. Two samples of essential oils were obtained from the aerial parts of the plant by hydrodistillation and analyzed by GC and GC-MS. The oils were characterized by their high content of monoterpenes (70.0-87.2 %). Important differences between the major constituents were found, particularly for carvacrol (0.2 vs. 44.8 %), thymol (18.1 vs. 7.4 %) and p-cymene (6.8 vs. 21.8 %). Both oils showed significant activity against all tested Gram-positive and Gram-negative bacteria, as well as against fungi. Nevertheless, the oil with the higher carvacrol content was found to be more active than the thymol type, with MICs ranging from 0.25 to 0.83 microL/mL and 0.12 to 0.27 microL/mL for bacteria and fungi, respectively. PMID- 12567289 TI - In vitro mass multiplication and production of roots in Plumbago rosea. AB - Mass multiplication of Plumbago rosea was achieved by indirect organogenesis in young stem, leaf and root explant cultures of 6-9-month-old plants. All the explants responded similarly in a hormonal regime of 2.5 mg/L BA and 1.5 mg/L NAA with the formation of nodular callus in 4 weeks; the callus was divided and subcultured at 4-week intervals in the presence of 3.0 mg/L BA to produce up to 23.5 +/- 1.6 shoots in 18 weeks and then at 2.0 mg/L BA to produce up to 79.6 +/- 1.5 shoots in 23 weeks. The shoots of 2.0-3.5 cm length were rooted easily in half-strength MS agar medium supplemented with 0.1 mg/L IBA and rooted plants established within 4 weeks at a 95-98 % rate without hardening. Eight weeks after establishment, the micropropagated plants were transferred to experimental plots and cultivated for 10 months to obtain a significantly higher number (18.0 +/- 0.5) of larger tuberous roots (137.4 +/- 3.4 g fw/plant) compared to conventional rooted cuttings (14.0 +/- 1.7, 47.9 +/- 1.6 g fw/plant). During this period, the concentration of the root-specific compound, plumbagin recorded per g dw (1.5 %), was higher than that of conventionally propagated plants (0.9-1.0 %). The early formation of plumbagin-rich tuberous roots holds significant potential for the commercial cultivation of the micropropagated plants. PMID- 12567290 TI - Micropropagation of Hydrastis canadensis: Goldenseal a North American endangered species. AB - An in vitro propagation protocol for rapidly producing Hydrastis canadensis L., Goldenseal, plantlets from disk tissue of young leaves was developed. Leaf explants were inoculated on MS medium supplemented with various concentrations of NAA and TDZ for production of callus. Two-month-old calli were sub-cultured on MS media containing cytokinins (BA, kinetin, TDZ) in different concentrations for shoot initiation. The optimum level of callus induction and maintenance was in 5.3 microM NAA in combination with 2.2 microM of TDZ. Shoot multiplication was achieved on MS medium with 2.2 microM TDZ in combination with 0.5 microM NAA. The alkaloid profile of micropropagated plantlets was similar to the profile of the mother plants. These results suggest that our in vitro propagation protocol will produce a positive impact in the conservation of H. canadensis. PMID- 12567291 TI - Polymeric proanthocyanidins from the bark of Hamamelis virginiana. AB - Polymeric proanthocyanidins were isolated from the bark of Hamamelis virginiana L. in yields of about 5 %. Fractionation yielded fractions with similar structures but different molecular weights with DP between 17-29 (thiolysis) and 11-20 (GPC). Polymers were composed predominantly of epicatechin and epigallocatechin as chain extension units at ratio of about 1.3:1. Terminal chain units were catechin (approximately 95 %) and gallocatechin (approximately 5 %). All chain extension units were completely galloylated at position O-3, while chain terminating units were not galloylated. Predominant interflavan linkages were 4 --> 8-bonds. PMID- 12567292 TI - New epoxyserratanes from Huperzia serrata. AB - Three new serratane-type epoxytriterpenoids were isolated from the whole plant of Huperzia serrata. The structures of these new compounds were characterized as 14 beta,15 beta-epoxy-3 beta-hydroxyserratan-21 beta-ol (1), 14 beta,15 beta-epoxy-3 beta-hydroxyserratan-21alpha-ol (2), and 14 beta,15 beta-epoxy-3 beta hydroxyserratan-21alpha-ol-3 beta-O-acetate (3), respectively, on the basis of spectral data and for 1 by single-crystal X-ray analysis. PMID- 12567293 TI - A new diepoxy-ent-kauranoid, rugosinin, from Isodon rugosus. AB - A new diterpenoid, rugosinin (1), isolated from Isodon rugosus, with absolute configuration was proved by single-crystal X-ray diffraction analysis, to be the member of a rare class of C-20/C-7 and C-20/C-14 diepoxy- ent-kauranoids. Effusanin A (2), effusanin B (3), effusanin E (4), lasiokaurin (5) and oridonin (6) were found as known constituents of the genus Isodon with C-20/C-7 epoxy function. These compounds have exhibited DNA-damaging activity in assay which employed DNA-repair deficient (RAD 52Y) and repair proficient (RAD +) yeast strains. PMID- 12567294 TI - Addition of a macrolide to a beta-lactam-based empirical antibiotic regimen is associated with lower in-hospital mortality for patients with bacteremic pneumococcal pneumonia. AB - To assess the association between inclusion of a macrolide in a beta-lactam-based empirical antibiotic regimen and mortality among patients with bacteremic pneumococcal pneumonia, 10 years of data from a database were analyzed. The total available set of putative prognostic factors was subjected to stepwise logistic regression, with in-hospital death as the dependent variable. Of the 409 patients analyzed, 238 (58%) received a beta-lactam plus a macrolide and 171 (42%) received a beta-lactam without a macrolide. Multivariate analysis revealed 4 variables to be independently associated with death: shock (P<.0001), age of >or=65 years (P=.02), infections with pathogens that have resistance to both penicillin and erythromycin (P=.04), and no inclusion of a macrolide in the initial antibiotic regimen (P=.03). For patients with bacteremic pneumococcal pneumonia, not adding a macrolide to a beta-lactam-based initial antibiotic regimen is an independent predictor of in-hospital mortality. However, only a randomized study can definitively determine whether this association is due to a real effect of macrolides. PMID- 12567295 TI - What is optimal antimicrobial therapy for bacteremic pneumococcal pneumonia? PMID- 12567296 TI - Respiratory tract infections in travelers: a review of the GeoSentinel surveillance network. AB - Respiratory tract infections are common in travelers, and improving our knowledge of risk factors associated with specific types of respiratory infections should enable implementation of better preventive strategies. Data collected by the GeoSentinel surveillance network were analyzed, and the most significant predictors for developing specific categories of respiratory infections while abroad were age, sex, season of travel, trip duration, and reason for travel. In particular, influenza was associated with travel to the Northern Hemisphere during the period of December through February, travel involving visits to friends or relatives, and trip duration of >30 days. Lower respiratory tract infections were associated with male sex and increasing age. Knowledge of the respiratory tract infections that occur in specific groups of travelers allows for the development of targeted pretravel preventive strategies to high-risk groups. PMID- 12567297 TI - Infection of transjugular intrahepatic portosystemic shunt devices: three cases and a review of the literature. AB - During the past decade, transjugular intrahepatic portosystemic shunting (TIPS) has become accepted as an alternative to the surgical shunt procedure for controlling the complications of portal hypertension, yet few data exist regarding infectious complications. The records of all patients who underwent a TIPS procedure during 1992-2000 at a single university teaching hospital were analyzed retrospectively to determine the rate and type of infections that occurred. A search of the English-language literature on the subject was conducted, and the findings were compared with our experience. Three cases of TIPS stent infection were identified out of 180 procedures performed during a 9 year period, resulting in an overall rate of infection of 1.7%. One of the patients was cured, and the other 2 died. A total of 21 cases of TIPS infection were identified from the literature search. A need exists for a standardized case definition for TIPS infection, and one is proposed. PMID- 12567298 TI - Factors associated with death among adults <55 years of age hospitalized for community-acquired pneumonia. AB - An administrative database was used to study death occurring among adults aged 18 55 years who were hospitalized during the period from 1 April 1994 through 31 March 1999 for treatment of community-acquired pneumonia. In-hospital case fatality rates for the first 10 days of hospitalization and overall were 2.1% and 3.2%, respectively, for 11,684 patient hospitalizations. Patient factors (age, sex, and comorbidity) were the most important associations with death. Aspiration provided the largest explanation of variance in deaths occurring during the first 10 days of hospitalization (odds ratio, 5.0; 95% confidence interval, 3.7-6.8). Busy hospitals (higher occupancy and higher number of daily admissions) were not associated with higher case-fatality rates. Bigger hospitals (metropolitan hospitals) had higher case-fatality rates, but this was more likely related to greater comorbidity and severity of pneumonia. Death due to community-acquired pneumonia among young and middle-aged adults is infrequent and is more related to the severity of pneumonia and to such risk factors as aspiration than to the manner in which the provision of care is organized. PMID- 12567299 TI - Serotype distribution, antibiotic susceptibility, and genetic relatedness of Neisseria meningitidis strains recently isolated in Italy. AB - The availability of new polysaccharide-protein conjugate vaccines against Neisseria meningitidis serogroup C prompted European National Health authorities to carefully monitor isolate characteristics. In Italy, during 1999-2001, the average incidence was 0.4 cases per 100,000 inhabitants. Serogroup B was predominant and accounted for 75% of the isolates, followed by serogroup C with 24%. Serogroup C was isolated almost twice as frequently in cases of septicemia than in cases of meningitis, and the most common phenotypes were C:2a:P1.5 and C:2b:P1.5. Among serogroup B meningococci, the trend of predominant phenotypes has changed from year to year, with a recent increase in the frequency of B:15:P1.4. Only a few meningococci had decreased susceptibility to penicillin, and, in the penA gene, all of these strains had exogenous DNA blocks deriving from the DNA of commensal Neisseria flavescens, Neisseria cinerea, and Neisseria perflava/sicca. Fluorescent amplified fragment-length polymorphism analysis revealed the nonclonal nature of the strains with decreased susceptibility to penicillin. PMID- 12567300 TI - Epidemiological and microbiological characterization of infections caused by Staphylococcus aureus with reduced susceptibility to vancomycin, United States, 1997-2001. AB - Infections caused by Staphylococcus aureus with reduced vancomycin susceptibility (SA-RVS; minimum inhibitory concentration [MIC], >or=4 microg/mL), including vancomycin-intermediate S. aureus (VISA; MIC, 8 microg/mL), are a new clinical and public health dilemma. Prospective surveillance and a nested case-control study of patients in the United States infected with SA-RVS was conduced from March 1999 through December 2000. Control patients were persons infected with oxacillin-resistant S. aureus (MIC of vancomycin, 20 serogroups undergo reaction with patient serum samples to detect agglutinating antibodies. Data derived from this assay are often used to infer the identity of the infecting leptospiral serovar or serogroup; however, paradoxical reactions and cross-reactions between serogroups are common. To evaluate the usefulness of this approach, data on culture-proven cases of leptospirosis that occurred in Barbados from January 1980 through December 1998 were reviewed. A total of 151 isolates of 4 serovars were identified. The sensitivity of MAT for the prediction of the infecting serovar was determined. Overall, the predominant serogroup at a titer of >or=100 correctly predicted 46.4% of all serovars isolated. If a titer of >or=800 was used as the cutoff, sensitivity decreased slightly to 44.4%. The overall specificity for all serogroups was 64.8%. Serologic analysis appeared to be of little value for the identification of the infecting serovar in individual cases of leptospirosis in humans. Presumptive serogroup reactivity data should be used only to gain a broad idea of the serogroups present at the population level. PMID- 12567303 TI - Nicotinamide: an oral antimicrobial agent with activity against both Mycobacterium tuberculosis and human immunodeficiency virus. AB - Coinfection with Mycobacterium tuberculosis and human immunodeficiency virus (HIV) is responsible for one-third of all deaths due to acquired immunodeficiency syndrome. More than 99% of cases of HIV-M. tuberculosis coinfection occur in the developing world, where limited resources add urgency to the search for effective and affordable therapies. Although antimicrobial agents against each of these infections are available, single agents that have activity against both M. tuberculosis and HIV are uncommon. The activity of nicotinamide has been evaluated in 2 different eras: in anti-M. tuberculosis studies performed during 1945-1961 and in anti-HIV studies performed from 1991 to the present. This review brings together these 2 bodies of inquiry and raises the possibility that, with more study, this small molecule could emerge at the beginning of the 21st century either as a therapeutic agent in itself or as the lead compound for a new class of agents with activity against both M. tuberculosis and HIV. PMID- 12567304 TI - Tetracycline therapy: update. AB - Tetracyclines have been used for treatment of a wide variety of gram-positive and gram-negative bacterial infections since the 1950s. In addition to being effective against traditional bacteria, tetracyclines have been used to treat infections due to intracellular chlamydiae, mycoplasmas, rickettsiae, and protozoan parasites and a variety of noninfectious conditions. They are important for treatment of and prophylaxis against infections with bacteria that could be used in biological weapons. Bacterial resistance to tetracycline was identified shortly after the introduction of therapy. At present, tetracycline resistance in bacteria can occur by acquisition of >or=1 of the 36 different genes, by mutations to host efflux pumps or in their 16S rRNA sequences, or by alteration in the permeability of the cell. In contrast, tetracycline resistance has not yet been described in protozoa or other eukaryotic organisms. PMID- 12567305 TI - Smallpox vaccination and patients with human immunodeficiency virus infection or acquired immunodeficiency syndrome. AB - Smallpox vaccination strategies are evolving rapidly and have important implications for human immunodeficiency virus (HIV)-infected persons. Cell mediated immunity is important for controlling both smallpox and vaccinia. For smallpox, the concern is a substantial increase in the associated mortality rate, which is 30% among healthy persons. For smallpox vaccination, the concern is progressive vaccinia, which is usually lethal but relatively uncommon. The risks associated with both smallpox and vaccinia viruses probably correlate with CD4 cell count, and, as a corollary, the best protection against infection with each is presumably immune reconstitution. It appears that all vaccinations will be voluntary, with 2 recommendations: (1) HIV-infected persons will be advised to decline preemptive vaccination, and (2) in the event of a bioterrorism attack involving smallpox, HIV-infected patients with exposures will be advised to receive vaccine. PMID- 12567306 TI - Quinupristin-dalfopristin and linezolid: evidence and opinion. AB - Quinupristin-dalfopristin and linezolid demonstrate in vitro activity against a wide range of gram-positive bacteria, including many isolates resistant to earlier antimicrobials. Quinupristin-dalfopristin is inactive against Enterococcus faecalis but has been effective for treatment of infections due to vancomycin-resistant Enterococcus faecium associated with bacteremia. In comparative trials, linezolid proved to be equivalent to comparator agents, resulting in its approval for several clinical indications. The almost-complete bioavailability of linezolid permits oral administration. Each agent can cause adverse effects that may limit use in individual patients. Resistance to these drugs has been encountered infrequently among vancomycin-resistant E. faecium. Resistance to quinupristin-dalfopristin is rare among staphylococci in the United States, and resistance to linezolid is very rare. Whether there is any benefit to use of these agents in combination regimens, and whether there are circumstances in which they might be alternatives to cell-wall active antibiotics for treatment of bone or endovascular infections, are questions that deserve further study. PMID- 12567307 TI - Longitudinal evolution of bone mineral density and bone markers in human immunodeficiency virus-infected individuals. AB - The underlying mechanisms of several bone disorders in human immunodeficiency virus (HIV)-infected persons and any relation to antiretroviral therapy have yet to be defined. A longitudinal study was conducted to estimate the prevalence of osteopenia or osteoporosis in HIV-infected persons; to assess bone mineralization, metabolism, and histomorphometry over time; and to evaluate predisposing factors. A total of 128 patients enrolled the study, and 93 were observed for 72 weeks. "Classic" risk factors (low body mass index, history of weight loss, steroid use, and smoking) for low bone mineral density (BMD) and duration of HIV infection were strongly associated with osteopenia. There was a weak association between low BMD and receipt of treatment with protease inhibitors; this association disappeared after controlling for the above factors. Markers of bone turnover tended to be elevated in the whole cohort but were not associated with low BMD. BMD increased slightly during follow-up. Traditional risk factors and advanced HIV infection play a more significant pathogenic role in the development of osteopenia and osteoporosis associated with HIV infection than do treatment-associated factors. PMID- 12567308 TI - Progression of chronic hepatitis C to liver fibrosis and cirrhosis in patients coinfected with hepatitis C virus and human immunodeficiency virus. AB - To evaluate the factors associated with the evolution of chronic hepatitis C in human immunodeficiency virus (HIV)-infected patients, a cross-sectional analysis of 41 HIV-infected patients with chronic hepatitis C (known as "HIV-HCV [hepatitis C virus]-coinfected patients") and a control group of patients with chronic hepatitis C who did not have HIV infection (known as "non-HIV-infected patients") was performed. The association of histological variables with demographic parameters, HCV load and genotype, HIV load, CD4(+) T cell count, and response to highly active antiretroviral therapy (HAART) was evaluated. HIV-HCV coinfected patients showed a significantly higher HCV load, more-advanced fibrosis, and a higher liver fibrosis progression rate (FPR) than did non-HIV infected patients. A high HCV load and a low CD4(+) T cell count were associated with a higher FPR. The immune response induced by HAART did not influence this progression. In conclusion, HIV-HCV-infected patients, mainly such patients with a high HCV load and an immunodepressed state, have a higher FPR. An independent effect of the immune response to HAART was not evident. PMID- 12567309 TI - Reduced testosterone levels in human immunodeficiency virus-infected women with weight loss and low weight. AB - Risk factors for hypoandrogenemia among low-weight, human immunodeficiency virus (HIV)-infected patients are not known. Testosterone levels of 69 HIV-infected women with low weight and weight loss were compared with levels for 25 healthy, age- and body mass index-matched control subjects. HIV-infected subjects were of low weight, with a mean (+/- standard deviation) weight loss of -17.6% +/- 9.7% from preillness maximum, and 42% of HIV-infected subjects had a body mass index of <20 kg/m(2). Forty-nine percent of the HIV-infected population versus 8% of the control population exhibited low free testosterone levels (P<.001). Among HIV infected women, when we controlled for chronic hepatitis status, age, and time of blood sampling, weight loss of >10% of maximum weight was a significant predictor of low free testosterone levels. Free testosterone levels did not differ by drug class or antiretroviral regimen. In conclusion, decreased androgen levels are common among HIV-infected women reporting significant weight loss, independent of exposure to antiretroviral medications. PMID- 12567310 TI - Dermatological findings of human T lymphotropic virus type 1 (HTLV-I)-associated myelopathy/tropical spastic paraparesis. AB - Dermatological findings for patients with human T lymphotropic virus type 1(HTLV I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) were investigated and were compared with dermatological findings for a control group. Only xerosis, cutaneous candidiasis, and palmar erythema were significantly associated with HAM/TSP. Histopathological patterns of cutaneous lymphoma were seen in 25% of 32 patients who underwent biopsy, and, thus, the cutaneous alterations in HAM/TSP can be classified into nonspecific lesions, infectious lesions, immune inflammatory-mediated lesions, and premalignant or malignant lesions. PMID- 12567311 TI - Knowledge of antiretroviral regimen dosing and adherence: a longitudinal study. AB - In a cohort of 128 human immunodeficiency virus-infected patients, we found that patients' knowledge of antiretroviral dosing was suboptimal at regimen initiation but improved with time. Poor medication knowledge 8 weeks after regimen initiation was associated with lower adherence and with a lower level of literacy in a multivariate model (P=.03). Because knowledge deficits are common after antiretroviral regimen initiation, clinicians should assess patients' understanding of medication dosing soon after regimen initiation or change. PMID- 12567312 TI - A case of plague successfully treated with ciprofloxacin and sympathetic blockade for treatment of gangrene. AB - A critically ill patient with septicemic plague and peripheral gangrene was treated successfully with ciprofloxacin. There are no previous reports of plague being successfully treated with ciprofloxacin. Peripheral gangrene of this patient's feet was managed with use of sympathetic blockade; the patient's toes appear to have been saved by this approach. PMID- 12567313 TI - Catheter-related bacteremia caused by the nocardioform actinomycete Gordonia terrae. AB - Five cases of catheter-related bacteremia caused by Gordonia terrae are reported. All patients who also had the primary diagnosis of cancer experienced nonneutropenic fever as a result of G. terrae infection. All patients were treated successfully with antibiotics, with the requirement of catheter removal for 2 patients who had systemic infections. PMID- 12567314 TI - Acute hepatitis B and C virus coinfection: a virological and clinical study of 3 cases. AB - We report the virological interaction in, clinical presentation of, and course of disease observed in 3 male injection drug users with acute hepatitis B virus (HBV) and hepatitis C virus (HCV) coinfection. In all 3 cases, HBV infection presented first and quickly resolved. Diagnosis of acute HBV/HCV coinfection requires a long follow-up period with careful observation. PMID- 12567315 TI - Infectious diarrhea in the southeastern United States, 1998-2000. PMID- 12567316 TI - A case of malaria from the Afghanistan region. PMID- 12567317 TI - Enteric fever in Mumbai, India: the good news and the bad news. PMID- 12567318 TI - Reduced acquisition of vancomycin-resistant enterococci: gown effect or confounding? PMID- 12567320 TI - Acetazolamide therapy and intracranial pressure. PMID- 12567321 TI - Pasteurella multocida urinary tract infection with molecular evidence of zoonotic transmission. AB - We describe a patient with a urinary tract infection (UTI) caused by Pasteurella multocida. Pulsed-field gel electrophoresis revealed that the clinical isolate recovered from the patient was identical (100% band match) to P. multocida isolates recovered from the patient's cat, but the isolate differed from an isolate recovered from a visiting cat and from a laboratory control strain. The patient also had abnormal urologic anatomy secondary to surgery; this has also been associated with P. multocida UTI. PMID- 12567322 TI - Clostridium difficile colitis after forced anal-receptive intercourse. AB - Colitis caused by Clostridium difficile toxin has been reported in homosexual men. We report a case of C. difficile colitis that occurred in an immunocompetent heterosexual man after forced anal-receptive intercourse. PMID- 12567323 TI - Successful treatment of Trichosporon mucoides infection with fluconazole in a heart and kidney transplant recipient. AB - Trichosporon species are an emerging cause of infection, particularly among transplant recipients. We report what we believe to be the first case of successful management of disseminated Trichosporon mucoides infection with orally administered fluconazole in a heart and kidney transplant recipient. PMID- 12567324 TI - Identification of a novel Bardet-Biedl syndrome protein, BBS7, that shares structural features with BBS1 and BBS2. AB - Bardet-Biedl syndrome (BBS) is a genetically heterogeneous disorder, the primary features of which include obesity, retinal dystrophy, polydactyly, hypogenitalism, learning difficulties, and renal malformations. Conventional linkage and positional cloning have led to the mapping of six BBS loci in the human genome, four of which (BBS1, BBS2, BBS4, and BBS6) have been cloned. Despite these advances, the protein sequences of the known BBS genes have provided little or no insight into their function. To delineate functionally important regions in BBS2, we performed phylogenetic and genomic studies in which we used the human and zebrafish BBS2 peptide sequences to search dbEST and the translation of the draft human genome. We identified two novel genes that we initially named "BBS2L1" and "BBS2L2" and that exhibit modest similarity with two discrete, overlapping regions of BBS2. In the present study, we demonstrate that BBS2L1 mutations cause BBS, thereby defining a novel locus for this syndrome, BBS7, whereas BBS2L2 has been shown independently to be BBS1. The motif-based identification of a novel BBS locus has enabled us to define a potential functional domain that is present in three of the five known BBS proteins and, therefore, is likely to be important in the pathogenesis of this complex syndrome. PMID- 12567328 TI - Seeking reversibility and treatability in dementia. AB - The search for "reversible causes" in the patient presenting with dementia has become a routine part of medical care. This article reviews evidence as to the prevalence of reversibility and concludes that actual reversal of dementia by treatment is rare. In this context, the utility of specific laboratory investigations is discussed, and commonly sought conditions, such as normal pressure hydrocephalus and B12 deficiency, are reviewed. Overemphasis on reversibility risks neglect of the assistance physicians can provide to patients suffering from irreversible conditions and their families. PMID- 12567325 TI - Fine mapping of autistic disorder to chromosome 15q11-q13 by use of phenotypic subtypes. AB - Autistic disorder (AutD) is a complex genetic disease. Available evidence suggests that several genes contribute to the underlying genetic risk for the development of AutD. However, both etiologic heterogeneity and genetic heterogeneity confound the discovery of AutD-susceptibility genes. Chromosome 15q11-q13 has been identified as a strong candidate region on the basis of both the frequent occurrence of chromosomal abnormalities in that region and numerous suggestive linkage and association findings. Ordered-subset analysis (OSA) is a novel statistical method to identify a homogeneous subset of families that contribute to overall linkage at a given chromosomal location and thus to potentially help in the fine mapping and localization of the susceptibility gene within a chromosomal area. For the present analysis, a factor that represents insistence on sameness (IS)--derived from a principal-component factor analysis using data on 221 patients with AutD from the repetitive behaviors/stereotyped patterns domain in the Autism Diagnostic Interview-Revised--was used as a covariate in OSA. Analysis of families sharing high scores on the IS factor increased linkage evidence for the 15q11-q13 region, at the GABRB3 locus, from a LOD score of 1.45 to a LOD score of 4.71. These results narrow our region of interest on chromosome 15 to an area surrounding the gamma-aminobutyric acid receptor subunit genes, in AutD, and support the hypothesis that the analysis of phenotypic homogeneous subtypes may be a powerful tool for the mapping of disease susceptibility genes in complex traits. PMID- 12567329 TI - Mild cognitive impairment: conceptual issues and structural and functional brain correlates. AB - Mild cognitive impairment (MCI) is a prevalent condition among older adults that carries a high risk of progression to Alzheimer's disease or other dementias. Given the potential for delaying or preventing the onset of dementia, efforts aimed at early detection and early intervention are important. The current paper reviews the conceptualization and diagnosis of MCI, assessment of memory complaints and deficits in the elderly, as well as recent research on the neurobiological basis of the disorder, including neurochemical, structural, and functional neuroimaging findings. PMID- 12567330 TI - Alzheimer's disease: from molecular biology to neuropsychiatry. AB - Alzheimer's disease is a clinical syndrome reflecting the topography and rate of progression of amyloid-associated cell death and dysfunction. Abnormal processing of amyloid precursor protein or enhanced accumulation of brain amyloid incites multiple pathogenetic pathway including inflammation, oxidation, and tau protein hyperphosphorylation that lead to neuronal death. The molecular biology of Alzheimer's disease provides insight into the clinical phenotype. The steps of the amyloid cascade present multiple targets for drug development and identification of effective disease-modifying anti-Alzheimer's disease treatments. PMID- 12567331 TI - Vascular dementia. AB - Vascular dementia (VaD) is a term used to describe a particular constellation of cognitive and functional impairment, and is now generally seen as a subset of the larger syndrome of vascular cognitive impairment (VCI). The latter is seen as cognitive impairment in the face of cerebrovascular disease. VCI can be classified clinically by whether patients meet criteria for dementia, and whether the syndrome is distinct or overlaps with primary neurodegenerative diseases, such as Alzheimer's disease. This clinical classification can be further classified by neuroimaging, with subgroups that show cortical infarction, subcortical infarction and white matter changes, each alone or in combination. Understood in this way, VCI is likely the most common form of cognitive impairment in the population. Attempts to treat VaD had varying degrees of success, but it now appears that many forms of VCI might be preventable, especially with good control of vascular risk factors in middle age. PMID- 12567332 TI - Dementia with Lewy bodies. AB - The objective was to summarize recent findings about the clinical features, diagnosis and investigation of dementia with Lewy (DLB) bodies, together with its neuropathology, neurochemistry and genetics. Dementia with Lewy bodies (DLB) is a primary, neurodegenerative dementia sharing clinical and pathological characteristics with both Parkinson's disease (PD) and Alzheimer's disease (AD). Antiubiquitin immunocytochemical staining, developed in the early 1990s, allowed the frequency and distribution of cortical LBs to be defined. More recently, alpha-synuclein antibodies have revealed extensive neuritic pathology in DLB demonstrating a neurobiological link with other "synucleinopathies" including PD and multiple system atrophy (MSA). The most significant correlates of cognitive failure in DLB appear to be with cortical LB and Lewy neurites (LNs) rather than Alzheimer type pathology. Clinical diagnostic criteria for DLB, published in 1996, have been subjected to several validation studies against autopsy findings. These conclude that although diagnostic specificity is high (range 79- 100%, mean 92%), sensitivity is lower (range 0- 83 %, mean, 49%). Improved methods of case detection are therefore required. Fluctuating impairments in attention, visual recognition and construction are more indicative of DLB than AD. Relative preservation of medial temporal lobe volume on structural MRI and the use of SPECT tracers for regional blood flow and the dopamine transporter are the most reliable current biomarkers for DLB. There are no genetic or CSF tests recommended for the diagnosis of DLB at present. Between 15 and 20% of all elderly demented cases reaching autopsy have DLB, making it the most common cause of degenerative dementia after AD. Exquisite, not infrequently fatal, sensitivity to neuroleptic drugs and encouraging reports of the effects of cholinesterase inhibitors on cognitive, psychiatric and neurological features, mean that an accurate diagnosis of DLB is more than merely of academic interest. Dementia developing late in the course of PD shares many of the same clinical and pathological characteristics. PMID- 12567333 TI - Frontotemporal dementias: clinical features and management. AB - This 2-part review of the frontotemporal lobar degeneration (FTLD) begins with an outline of clinical features, which differentiate FTLD from Alzheimer's disease, the more common cause of dementia. The second part describes interventions for FTLD. Although there is currently no disease-modifying therapeutic agent, symptomatic pharmacotherapy helps to control the marked mood and behavioral disturbances that may distinguish this syndrome from other causes of dementia. PMID- 12567334 TI - The classic and variant forms of Creutzfeldt-Jakob disease. AB - Transmissible spongiform encephalopathies (TSEs), also known as prion diseases, are a group of progressive neurodegenerative disorders that affect both animals and humans. These diseases take part of their name from a striking and common neuropathological feature: spongiform ("sponge-like") degeneration of the brain. As the name also implies, TSEs can literally be transmitted from one host to another much like any other infectious disease. In these situations, however, symptoms may take months or years to appear in the recipient host. Otherwise, TSEs either develop spontaneously or rarely arise through genetic mutation and therefore occur as familial disorders. Regardless of their origin, all TSEs progress over a period of months inevitably leading to the death of the affected individual. From a neuropsychiatric perspective, it is notable that the newly identified variant form of Creutzfeldt-Jakob disease (vCJD) typically occurs in younger adults causing prominent psychiatric and/or behavioral abnormalities as important early manifestations of disease. This has forced a heightened awareness of vCJD among mental health professionals throughout the United Kingdom where the disorder occurs. Relevant features of both the classic and variant forms of Creutzfeldt-Jakob disease (CJD) will be reviewed. PMID- 12567335 TI - Evidence-based reports--is there any evidence? PMID- 12567336 TI - Central nervous system and cardiac effects from long-acting amide local anesthetic toxicity in the intact animal model. AB - With the development of the newer long-acting amide local anesthetics,ropivacaine and levobupivacaine, numerous animal studies of LA systemic toxicity have emerged. Because of the complex nature of the human response to LA intoxication, the task of designing and interpreting these animal studies of LA toxicity can be difficult. Accordingly, this report will review the selection of an animal model for the study of LA toxicity; examine the pertinent in vivo animal studies that compare the central nervous system toxicity, cardiovascular toxicity, and the ease of resuscitation of the single enantiomer local anesthetics to racemic bupivacaine; and extrapolate these findings to the clinical setting. PMID- 12567337 TI - Ketamine stimulates secretion of beta-endorphin from a mouse pituitary cell line. AB - BACKGROUND AND OBJECTIVES: beta-endorphin is an endogenous opioid that mediates pain-induced analgesia. Propofol inhibits in vitro secretion of beta-endorphin from a mouse pituitary cell line (AtT-20). We hypothesized that ketamine would also alter secretion of beta-endorphin. METHODS: AtT-20 cells were exposed to the intravenous anesthetic ketamine (10 to 40 micromol/L). Secretion of beta endorphin was determined by enzyme-linked immunosorbent assay. Long-term effects were determined by exposing the cells to ketamine, allowing the cells to recover overnight, then stimulating the secretion of beta-endorphin. AtT-20 cells were stimulated with secretagogues to induce secretion of beta-endorphin. The effect of ketamine on stimulated secretion was determined. Cultures of AtT-20 cells were grown for 5 days in the presence of ketamine. Cell numbers were determined on each day. RESULTS: Ketamine increased secretion of beta-endorphin to levels that were up to 3 times greater than baseline secretion. Stimulation of beta-endorphin secretion by ketamine persisted into the subsequent day. Ketamine caused increased secretion from cells stimulated with secretagogues. Ketamine was not toxic to these cells; AtT-20 cells grew normally for 5 days in the presence of up to 40 micromol/L ketamine. CONCLUSIONS: Clinically relevant concentrations of ketamine stimulated both immediate and delayed secretion of beta-endorphin. This suggests that the prolonged analgesia observed in some clinical situations with ketamine could be in part caused by increased release of an endogenous opioid. PMID- 12567338 TI - Delivery rate accuracy of portable, bolus-capable infusion pumps used for patient controlled continuous regional analgesia. AB - BACKGROUND AND OBJECTIVES: Portable patient-controlled pumps used for perineural local anesthetic infusion appear to be gaining acceptance and are used for an increasing number of medically unsupervised ambulatory patients. The infusion rate accuracy and reliability of these pumps, which allow patient-controlled boluses of potentially toxic medication, has not been independently investigated. We performed this laboratory study to define the flow-rate accuracy, reliability, and profiles of various portable bolus-capable infusion pumps and varied several factors that may influence pump performance. METHODS: Using a computer/scale combination, 4 electronic pumps were tested with 1 and 2 sets of batteries over 100 hours. One elastomeric pump was tested with its flow-regulator at room temperature (20 degrees C to 24 degrees C), skin temperature (32 degrees C), and increased temperature (36 degrees C). The elastomeric pump was also tested by varying the height of the pump relative to the distal end of the catheter. RESULTS: Infusion rate accuracy differed significantly among the pumps, exhibiting flow rates within +/-15% of their expected rate for 29% to 100% of their infusion duration. Changing the batteries after 50 hours affected the profile of 1 of the 4 electronic pumps. For the elastomeric pump, an increase in temperature significantly increased the infusion rate. An increase or decrease in pump-to-scale height significantly increased or decreased the infusion rate, respectively. CONCLUSIONS: These results suggest that factors such as flow-rate accuracy and consistency, infusion profile, and temperature sensitivity should be taken into consideration when choosing a portable infusion pump for local anesthetic administration. PMID- 12567339 TI - Effect of bupivacaine with epinephrine wound instillation for pain relief after pediatric inguinal herniorrhaphy and hydrocelectomy. AB - BACKGROUND AND OBJECTIVES: To evaluate the effect of a 20- or 60-second instillation period using 0.5% bupivacaine with epinephrine for pain relief after pediatric inguinal herniorrhaphy and hydrocelectomy. METHODS: In a randomized, double-blind study, 103 children (aged 1 to 12 years, American Society of Anesthesiologists [ASA] physical status I or II) were allocated into 4 groups after induction of anesthesia. Group 1: normal saline 0.25 mL/kg instilled, which remained in the wound for 20 or 60 seconds before wound closure. Group 2: 0.25 mL/kg 0.5% bupivacaine with epinephrine 5 microg/mL instilled, which remained in the wound for 20 seconds. Group 3: the same quantity and dose of drug 2 instilled as group, but remained in the wound for 60 seconds. Group 4: an ilioinguinal and iliohypogastric block performed before operation using 0.5 mL/kg 0.25% bupivacaine with epinephrine. The Children's Hospital of Eastern Ontario Pain Scale (CHEOPS) and Aldrete-Kroulik recovery scores were used to monitor postoperative pain and recovery status. Analgesic was given when the CHEOPS score was >/= 7 despite other supportive therapy. RESULTS: The number of patients requiring analgesics within 2 hours in group 1 (73.1%) was more than groups 2, 3, and 4 (23.1%, 20.8%, and 16%, respectively, P <.001). The median time to first analgesic in group 1 (50 minutes) was also less than groups 2, 3, and 4 (420, 525, and 425 minutes, respectively, P <.0001). CONCLUSION: 0.5% Bupivacaine with epinephrine for as short an instillation period as 20 or 60 seconds can provide a good analgesic alternative after herniorrhaphy and hydrocelectomy in pediatric patients. All studied blocks had comparable duration of action. PMID- 12567340 TI - Femoral block provides superior analgesia compared with intra-articular ropivacaine after anterior cruciate ligament reconstruction. AB - BACKGROUND AND OBJECTIVES: Arthroscopic anterior cruciate ligament (ACL) reconstruction of the knee is a painful procedure requiring intensive postoperative pain management. This prospective study investigates analgesic quality after a femoral block as compared with intra-articular injection of local anesthetic. METHODS: Eighty patients scheduled for elective ACL repair under general anesthesia were included in our study. Upon completion of surgery, the patients were randomly assigned into 1 of 2 groups: femoral group (n = 40) received a femoral block with 20 mL 1% ropivacaine; intra-articular group (n = 40) received 20 mL 1% ropivacaine injected intra-articularly. During the first 24 hours after surgery, all patients received 2 g propacetamol and 100 mg ketoprofen, intravenously. Additional postoperative analgesia was available with parenteral morphine if required. Analgesic duration was defined as the time from end of surgery to the first requirement for a supplemental analgesic. Data collection included patient demographics, visual analog scale (VAS) scores, analgesic duration, and morphine use. Analysis of variance (ANOVA) test was used to compare the 2 groups. RESULTS: VAS score in the recovery room and during rehabilitation was higher in the intra-articular group than in the femoral group (P <.001). Morphine use was lower in the femoral group than in the intra articular group (P <.001). Similarly, analgesic duration was longer in the femoral group than the intra-articular group (P <.0001). CONCLUSIONS: Compared with intra-articular injection of local anesthetic, femoral nerve block (FNB) provides better analgesia and allows a significant morphine-sparing effect after ACL repair. PMID- 12567341 TI - Restricted infraclavicular distribution of the local anesthetic solution after infraclavicular brachial plexus block. AB - BACKGROUND AND OBJECTIVES: The distribution of local anesthetic after different approaches for brachial plexus anesthesia could be responsible for the varying rates of side effects, such as phrenic block, hoarseness, and Horner's syndrome associated with each approach. We compared the distribution of local anesthetic within the neurovascular space in infraclavicular block with that of interscalene and supraclavicular block. METHODS: In a prospective analysis using fluoroscopy, we studied the distribution of a solution of local anesthetic containing radiologic contrast medium in 18 patients. Six patients received an interscalene block, another 6 patients received a perpendicular supraclavicular block, and another 6 patients, a perpendicular coracoid block. RESULTS: Distribution of the anesthetic solution in the interscalene and supraclavicular groups extended to both supraclavicular and infraclavicular spaces in all patients. This distribution was significantly different (P <.05) compared with that of the infraclavicular group. In this group, the solution remained below the clavicle in every patient. CONCLUSIONS: Spread of the local anesthetic from the infraclavicular space after infraclavicular coracoid block appears to be limited to below the level of the clavicle. Conversely, local anesthetic solution passes below the clavicle in all patients given interscalene or supraclavicular blocks. PMID- 12567342 TI - Variations in epidural catheter manufacture: implications for bending and stiffness. AB - BACKGROUND AND OBJECTIVES: There is no formal evaluation method used to relate epidural catheter design and manufacture to clinical outcomes, such as subarachnoid or intravascular catheter placement. We analyzed catheter bending stiffness to determine the range of stiffness of catheters commonly used. We hypothesized that catheter material has a greater influence on stiffness than does cross-sectional shape. METHODS: We determined the elastic modulus by axial load testing and the area moment of inertia using calibrated microscopic measurements of cross-sectional geometry for 6 different catheter types, including 2 types of wire styletted catheters. We calculated bending stiffness as the product of the elastic modulus and the area moment of inertia. RESULTS: Catheters had similar area moments of inertia, but markedly different elastic moduli. Nylon and polyurethane catheters had the same bending stiffness, which was twice as high as that of coil reinforced catheters (P <.05), but 35% lower than that of radiopaque catheters (P <.05). Nylon and radiopaque wire styletted catheters had similar bending stiffness, which were 23-fold to 90-fold greater than that of the nonstyletted catheters (P <.05). CONCLUSIONS: Catheters currently available establish the range of bending stiffness that should not be exceeded, only optimized to clinical outcome. Clinical studies are needed to correlate the incidence of unintentional intravascular or subarachnoid catheter placement or migration and bending stiffness. Catheter technology improvements may enhance safety and increase the likelihood of successful catheter insertion, maintenance, and removal. PMID- 12567344 TI - Assessing the superiority of saline versus air for use in the epidural loss of resistance technique: a literature review. PMID- 12567343 TI - Usefulness of pre-emptive peribulbar block in pediatric vitreoretinal surgery: a prospective study. AB - BACKGROUND AND OBJECTIVES: Vitreoretinal (VR) surgery with or without scleral buckling is associated with significant postoperative pain and emesis in adults, and recent studies have addressed the effect of retro or peribulbar block on these parameters. VR surgery in children has received little attention regarding the incidence of pain and emesis, and the role of regional anesthesia in modifying these parameters. In this study, we compared peribulbar block with conventional opioid analgesia in children undergoing VR surgery. METHODS: In a prospective, randomized, single-blind study, 85 children (ages 6 to 13 years) were allocated to receive peribulbar block (n = 42) or intravenous meperidine 1 mg/kg (n = 43) after induction of general anesthesia. Parameters compared were: intraoperative incidence of oculocardiac reflex and requirement for additional analgesic; postoperative pain intensity; incidence of postoperative emesis; time to first analgesic, total number of postoperative analgesic supplements; and parental assessment of the child's postoperative comfort at 24 hours. RESULTS: The incidence of intraoperative oculocardiac reflex was significantly less in the peribulbar group (P =.0001). Significantly more children receiving peribulbar block were pain free on awakening (P =.0004) and throughout the postoperative period. The number of children requiring opioid was significantly lower with peribulbar block (P =.008), and a significant number of children did not vomit throughout the postoperative period (P =.001). CONCLUSIONS: Peribulbar block appears to be a safe and clinically superior alternative to intravenous opioid for pediatric VR surgery. PMID- 12567345 TI - The management of nonobstetric pains in pregnancy. AB - OBJECTIVE: Nonobstetric pain in pregnancy is usually managed with simple analgesia and reassurance that the condition will be self-limiting. However, sometimes such pain can be severe enough to warrant a different approach. We present 3 cases of uncontrolled pain in pregnancy, which were treated with nerve block techniques. CASE REPORT: The first patient had a recurrence of pain similar to that experienced during her first pregnancy related to stretching of her lower ribs; this was relieved completely by injection of bupivacaine 0.5% and triamcinolone. Another patient had symphyseal pain, which had required induction of labor during her previous pregnancy due to its severity. She had only brief relief on injection. The third patient experienced severe pain related to a Pfannenstiel incision scar from a previous cesarean delivery, which was completely relieved by injection. A review is presented of nonobstetric pains and their treatment options that may occur in pregnancy, including neurogenic, musculoskeletal, neuropathic, and referred pains. CONCLUSION: We conclude that a single bolus injection of local anesthetic and steroid, when performed carefully by a pain management specialist, is a safe and potentially effective alternative to conventional analgesic treatment of somatic pain associated with the uterine enlargement of pregnancy. PMID- 12567346 TI - Anesthesia for outpatient knee arthroscopy: is there an optimal technique? PMID- 12567347 TI - Superior sagittal sinus thrombosis in a patient with postdural puncture headache. AB - BACKGROUND AND OBJECTIVES: The occurrence of concomitant intracranial pathology in a patient with postdural puncture headache (PDPH) is rare. We present a patient who had a superior sagittal sinus thrombosis in addition to his PDPH. The signs and symptoms of intracranial pathology in patients with dural puncture headache, in addition to their postural headache, are discussed. CASE REPORT: A 32-year-old man with lymphoblastic lymphoma received treatment with daunorubicin, vincristine, and prednisone. He developed postural headache and severe nausea and vomiting after a diagnostic lumbar puncture. Magnetic resonance imaging (MRI) showed superior sagittal sinus (SSS) thrombosis and meningeal enhancement. An epidural blood patch was performed and enoxaparin was prescribed for 6 months. He has remained asymptomatic. CONCLUSIONS: Patients with PDPH have classic postural headache. The occurrence of additional signs and symptoms should alert the clinician to the presence of intracranial pathology. Patients with lymphoblastic lymphoma who had treatment with L-asparaginase and steroid are predisposed to the development of cortical venous thrombosis and may have this syndrome in addition to a dural puncture headache. PMID- 12567348 TI - Splanchnic neurolysis using carbon dioxide as the contrast agent. AB - BACKGROUND AND OBJECTIVE: Iodinated contrast agents are usually used to verify correct spread of injectate during splanchnic neurolysis. We performed a splanchnic neurolysis by using carbon dioxide as the contrast agent in a patient who was allergic to iodinated contrast agents. CASE REPORT: A 49-year-old man had severe upper abdominal and back pain due to chronic pancreatitis. Because slow release morphine 360 mg a day and epidural fentanyl 500 microg a day did not relieve the pain, a splanchnic neurolysis was performed by the posterior approach. The needle tip was placed into the retrocrural space under fluoroscopic guidance. Fifteen milliliters of carbon dioxide was injected as the contrast agent. It gave a less clear image than that obtained by iodinated contrast agents; however, the correct spread of the gas was easily visible under continuous fluoroscopy. Six percent aqueous phenol 8 mL was injected after confirming relief of the pain with a local anesthetic. The dose of slow-release morphine was gradually decreased after the procedure. His abdominal and back pain was controlled with slow-release morphine 120 mg a day. CONCLUSION: Carbon dioxide was useful as the contrast agent to perform splanchnic neurolysis. PMID- 12567349 TI - Epidural fibrin glue injection stops postdural puncture headache in patient with long-term intrathecal catheterization. PMID- 12567350 TI - Continuous brachial plexus block using the posterior approach. PMID- 12567351 TI - Seizures after epidural blood patch. PMID- 12567353 TI - Fascicular anatomy, nervi nervorum, and paresthesia. PMID- 12567355 TI - Replacement of spinal cord stimulation electrode. PMID- 12567356 TI - Cerebrospinal fluid polyamine levels in patients with colonic disease. PMID- 12567357 TI - A British comment on the King's English. PMID- 12567361 TI - A radiographic study of the relationship between metatarsus adductus and hallux valgus. AB - The relationship between metatarsus adductus and hallux valgus was evaluated on 100 dorsoplantar weight-bearing radiographs. The metatarsus adductus angle varied positively with the degree of hallux valgus. By using measures of correlation, a significant linear association was found for women (r =.53, P <.001) and men (r =.48, P <.001). The relationship was strongest in women when all cases of abnormal metatarsus adductus (>24 degrees ) were associated with abnormal degrees of hallux valgus (>15 degrees ). This relationship was different in men in that abnormal metatarsus adductus angles were not always associated with abnormal hallux valgus angles. With male and female data combined, the prevalence of metatarsus adductus was 55% in subjects with hallux valgus deformity compared with 19% in subjects without hallux valgus. A Chi(2) test showed this to be a significant difference in the distribution of the data (P =.002). The data of this study suggests that there may be a clinical association between metatarsus adductus and hallux valgus. The need to further evaluate the role of metatarsus adductus angle in hallux valgus surgery is emphasized. PMID- 12567360 TI - Rapid diagnosis of pedal osteomyelitis in diabetics with a technetium-99m-labeled monoclonal antigranulocyte antibody. AB - An in vivo method of labeling white cells that diagnoses diabetic pedal osteomyelitis safely, rapidly, and accurately is desirable. The objectives of this investigation were to evaluate a technetium-99m-labeled monoclonal antigranulocyte antibody for diagnosing diabetic pedal osteomyelitis, compared with indium-111-labeled leukocyte and 3-phase bone imaging for this purpose. Twenty-five diabetic patients with pedal ulcers, 22 in the forefoot and 3 in the midfoot, underwent antibody, indium-111-labeled leukocyte, and technetium-99m methylene diphosphonate 3-phase bone imaging. The 1-hour antibody, 24-hour labeled leukocyte, and 3-phase bone images were interpreted separately for the presence of osteomyelitis. The antibody and labeled leukocyte images also were interpreted together with the bone images to determine if the combined study was more accurate than each individual study. There were 10 cases of osteomyelitis among the 25 patients. The sensitivity, specificity, and accuracy of the antibody were.90,.67, and.76, respectively. These results were not significantly different from those obtained with labeled leukocyte imaging:.80,.67, and.72, respectively. The antibody was significantly more specific (P =.004) than 3-phase bone imaging (.27). Interpreting the antibody together with the bone scan did not alter the results. When interpreted with the bone images, the accuracy of labeled leukocyte imaging improved from.72 to.80. This was not significantly more accurate than either the antibody or labeled leukocyte imaging alone. The data suggest that the monoclonal antigranulocyte antibody is comparable with in vitro labeled leukocyte imaging for diagnosing pedal osteomyelitis in diabetic patients, and warrants further investigation in a larger population. PMID- 12567362 TI - Mechanical and structural characteristics of the new BONE-LOK cortical-cancellous internal fixation device. AB - The purpose of this study was to evaluate the structural and mechanical characteristics of a new and unique titanium cortical-cancellous helical compression anchor with BONE-LOK (Triage Medical, Inc., Irvine, CA) technology for compressive internal fixation of fractures and osteotomies. This device provides fixation through the use of a distal helical anchor and a proximal retentive collar that are united by an axially movable pin (U.S. and international patents issued and pending). The helical compression anchor (2.7-mm diameter) was compared with 3.0-mm diameter titanium cancellous screws (Synthes, Paoli, PA) for pullout strength and compression in 7# and 12# synthetic rigid polyurethane foam (simulated bone matrix), and for 3-point bending stiffness. The following results (mean +/- standard deviation) were obtained: foam block pullout strength in 12# foam: 2.7-mm helical compression anchor 70 +/- 2.0 N and 3.0-mm titanium cancellous screws 37 +/- 11 N; in 7# foam: 2.7-mm helical compression anchor 33 +/- 3 N and 3.0-mm titanium cancellous screws 31 +/- 12 N. Three-point bending stiffness, 2.7-mm helical compression anchor 988 +/- 68 N/mm and 3.0-mm titanium cancellous screws 845 +/- 88 N/mm. Compression strength testing in 12# foam: 2.7-mm helical compression anchor 70.8 +/- 4.8 N and 3.0-mm titanium cancellous screws 23.0 +/- 3.1 N, in 7# foam: 2.7-mm helical compression anchor 42.6 +/- 3.2 N and 3.0-mm titanium cancellous screws 10.4 +/- 0.9 N. Results showed greater pullout strength, 3-point bending stiffness, and compression strength for the 2.7-mm helical compression anchor as compared with the 3.0-mm titanium cancellous screws in these testing models. This difference represents a distinct advantage in the new device that warrants further in vivo testing. PMID- 12567363 TI - Os calcis fractures: analysis of interobserver variability in using Sanders classification. AB - The os calcis is the most frequently fractured tarsal bone. In 1992 Sanders developed a classification system based on coronal and axial computed tomography (CT) scans of the calcaneus. This classification is the one used most frequently today in treatment decision making and reporting of results. The objective of this study was to assess the degree of interobserver variability in using this classification system. Thirty CTs of calcaneal fractures were chosen randomly from the past 5 years in 2 tertiary care centers. The CTs were reviewed by 3 orthopedic surgeons and one senior orthopedic resident who classified the fractures according to Sanders' classification. The results were first tabulated and analyzed by using a weighted kappa test including the subcategories. The weighted kappa value achieved was.56, with a 95% confidence interval of.45-.67. The subcategories of the classification were then further combined and a second weighted kappa test was performed to assess agreement between general classes. The weighted kappa value achieved was.48, with a 95% confidence interval of 0.37 0.59. We concluded that Sanders' classification system did prove to achieve moderate agreement among users, thus representing a useful classification system. PMID- 12567364 TI - The use of a local anesthetic foot block in patients undergoing outpatient bony forefoot surgery: a prospective randomized controlled trial. AB - Foot blocks are known to prolong postoperative pain relief. Consequently, their use has been extended to patients having outpatient surgery, despite little evidence to show improved patient satisfaction. Indeed, patients having outpatient surgery actually may be less satisfied because they will first experience pain at home on the first postoperative night, which may be more severe than anticipated. In this prospective, randomized, blinded study, 42 patients having unilateral outpatient bony forefoot surgery were assigned randomly to either having the surgery under general anesthesia alone or under general anesthesia with supplementary foot block (0.5% bupivicaine). All patients were assessed at home by telephone interview on the first and second postoperative day. There was a significantly longer time to first perceived pain in the foot block group compared with the control group, but no difference in the number of postoperative analgesic tablets consumed, no difference in pain score on the first night, first postoperative or second postoperative day, or any difference in the overall patient satisfaction scores at 2 days. The authors conclude that a local foot block, although prolonging the time to first perceived pain, does not improve patient satisfaction and is not detrimental when used as analgesia in the outpatient setting. PMID- 12567365 TI - Effect of noncontact normothermic wound therapy on the healing of neuropathic (diabetic) foot ulcers: an interim analysis of 20 patients. AB - This is the interim analysis of a prospective, randomized, controlled study comparing diabetic foot ulcer healing in patients being treated with either noncontact normothermic wound therapy (Warm-UP; Augustine Medical Inc. Eden Prairie, MN) applied for 1 hour 3 times daily until healing or 12 weeks, or standard care (saline-moistened gauze applied once a day). Surgical debridement and adequate foot off-loading was provided to both groups. Evaluations were performed weekly and consisted of acetate tracings, wound assessment, and serial photography. Twenty patients have completed the trial and both treatment groups were distributed evenly (N = 10). Ulcers treated with noncontact normothermic wound therapy had a greater mean percent wound closure than control-treated ulcers at each evaluation point (weeks 1-12). After 12 weeks, 70% of the wounds treated with noncontact normothermic wound therapy were healed compared with 40% for the control group. In this subset of patients there have been no adverse events associated with noncontact normothermic wound therapy. PMID- 12567366 TI - Long oblique distal osteotomy of the fifth metatarsal for correction of tailor's bunion: a retrospective review. AB - A long-term retrospective review of a long oblique distal osteotomy of the fifth metatarsal for correction of a tailor's bunion deformity is presented. The technique, results, and follow-up of this procedure are described. The long oblique distal osteotomy was performed on 25 feet on 16 patients. Follow-up ranged from 11 to 110 months with an average of 64.4 months. The average preoperative intermetatarsal 4-5 angle and lateral deviation angle improved from 10.88 degrees and 4.24 degrees, respectively, to 5.52 degrees and -4.24 degrees postoperatively, respectively. A modified scoring scale for lesser metatarsal procedures was used, showing an average postoperative score of 94 out of 100 possible points. Subjectively, 96% (24 of 25) of feet that underwent surgery were rated as excellent or good. Complications included one patient with paresthesias, one soft-tissue infection, and 2 patients had irritation from screws. The authors conclude that the long oblique distal osteotomy is a viable alternative in the correction of tailor's bunion deformity. PMID- 12567367 TI - Bilateral fractures of the infracalcaneal exostosis. AB - A case study of bilateral fractures of the inferior calcaneal spur is provided. The patient healed uneventfully after immobilization, cessation of activities, and oral medication. Thirteen months after initial presentation she remains asymptomatic. A brief literature review and discussion of the possible mechanism(s) of injury is included. PMID- 12567368 TI - Recurrent ankle sprains secondary to nonunion of a lateral malleolus fracture. AB - A case of an adult man with symptoms of chronic recurrent ankle sprains secondary to nonunion of a fracture of the tip of the lateral malleolus is presented. The nonunion was debrided, bone grafted, and internally fixed by using the tension band wire technique. The fracture healed and the patient experienced no further episodes of ankle sprain. PMID- 12567369 TI - The crescentic inverted scarf osteotomy. AB - A hybrid procedure combining the inverted scarf and crescentic osteotomies is presented. This procedure may serve as a viable modification of the inverted scarf bunionectomy for those surgeons desiring true rotational correction of high intermetatarsal angles. PMID- 12567370 TI - Stories and the brain: making crucial connections. PMID- 12567371 TI - Her records. PMID- 12567372 TI - Assessing risk for depression among immigrants at two-year follow-up. AB - This article reports findings from a longitudinal study of what predicts depression 2 years later in a sample of former Soviet immigrants. Study aims were to: (1) investigate change in depression scores and (2) determine the relative contribution of initial depression scores, select demographic and migration characteristics and common demands of immigration to depression at 2-year follow up. Data were collected at two time points from 468 former Soviet immigrants who had been in the US 5 years or less at the time of the first data collection appointment. Linear regression was used to predict depression at 2-year follow up. There was little improvement in depression for the sample as a whole over time. Baseline depression, unemployment, novelty, and not having relatives in the local resettlement area were significant predictors of depression at 2-year follow-up. Initial depression in relatively recent immigrants should be treated aggressively and not assumed to be a transient problem. Immigrants should be assisted to find early employment and overcome novel aspects of their new environment. Immigrants without local families will likely need additional support. PMID- 12567374 TI - Health promotion information: sources and significance for those with serious and persistent mental illness. AB - For 19 sources of health promotion information 41 adult outpatients with severe and persistent mental illness (SPMI) indicated by questionnaire how much information they received from each source and their opinion of its reliability. Nonpsychiatrist physicians, psychiatrists, nurses and pharmacists provided the most health promotion information and were regarded as the most reliable sources. Nurses and pharmacist were seen as providing more, and media and print sources as providing less, information than in similar studies of nonmentally ill samples. Nonpsychiatrist physicians, psychiatrists, nurses and pharmacists are significant sources of health promotion information for persons with SPMI--perhaps even more so than for those with no mental illness. PMID- 12567373 TI - Stress and coping in rural women with alcohol and other drug disorders. AB - Alcohol and other drug disorders (AOD) are a growing problem among women with estimates as high as 11 million U.S. women affected. This is a report of the results of two studies investigating risk factors for AOD disorders in rural women, a population that is difficult to access. Samples of the two studies were combined for a total of 267 women. Analyses focus on stress, coping, alcohol expectancies, and self-esteem. Results indicate that rural women with AOD disorders experience more stressors and fewer uplifting events, have fewer coping resources, and use more emotion-focused coping strategies than women without AOD disorders. PMID- 12567375 TI - Mitochondrial DNA evidence for admixed origins of central Siberian populations. AB - The Yakuts of northeastern Siberia are a Turkic-speaking population of horse- and cattle-breeders surrounded by Tungusic-speaking reindeer-herders and hunter gatherers. Archaeological and ethnohistorical data suggest that Yakuts stem from a common ancestral population with the Buryats living near Lake Baikal. To address this hypothesis, we obtained sequences of the first hypervariable segment (HV1) of the mitochondrial DNA control region from Yakuts and Buryats and compared these with sequences from other Eurasian populations. The mtDNA results show that the Buryats have close affinities with both Central Asian Turkic groups and Mongols, while the Yakuts have close affinities with northeastern Siberian, Tungusic-speaking Evenks and south Siberian, Turkic-speaking Tuvans. This different ancestry of the Yakuts and the Tuvans (compared with other Turkic speaking groups) most likely reflects extensive admixture that occurred between Turkic-speaking steppe groups and Evenks as the former migrated into Siberia. Moreover, the Yakuts are unique among Siberian populations in having a high number of haplotypes shared exclusively with Europeans, suggesting, contrary to the historical record, that occasionally Yakut men took Russian women as wives. PMID- 12567376 TI - Nonalimentary tooth use in prehistory: an example from early Holocene in Central Sahara (Uan Muhuggiag, Tadrart Acacus, Libya). AB - Signs of nonalimentary tooth use were observed on the dentition of an adult male from a single burial excavated in an area close to the Uan Muhuggiag rock shelter (Tadrart Acacus, Libya), dated to more than 7800 uncalibrated years BP, that represents the most ancient human remain found in the Libyan Sahara, and provides a first glimpse of human adaptation in the early Holocene of this region. The wear pattern shows large grooves running across the occlusal surfaces of maxillary and mandibular anterior teeth and premolars. The results of macroscopic and microscopic observation, together with scanning electron microscope (SEM) examination and experimental tests, suggest that the microdamage might be due to repeated friction of vegetal fibers, probably as a consequence of basket making, net production, or mat processing. Further data are needed to allow us to distinguish among plant-oriented activities related to food acquisition (e.g., rope and net processing), food storage (e.g., basket making), or domestic handicraft (e.g., mat processing), whose implications may generate different interpretations of sexual division of labor. PMID- 12567377 TI - Estimating the length of incomplete long bones: forensic standards from Guatemala. AB - We report on new standards for estimating long bone length from incomplete bones for use in forensic and archaeological contexts in Central America. The measurements we use closely follow those defined by Steele ([1970] Personal Identification in Mass Disasters; Washington, DC: Smithsonian Institution), but we add several new landmarks. We measured the femur, humerus, tibia, and fibula of 100 Maya skeletons (68 males, 32 females) recovered from forensic exhumations. We derived the equations by regressing bone segment length on bone length, and solved for bone length to maximize the utility of the equations for taller populations. We generated equations for all segments that were significantly correlated with bone length for males, for females, and for both sexes combined, but accepted only regressions with r(2) > 0.85 as reliable. Landmarks defined by muscle attachment sites were more variable in location than landmarks on articular architecture; thus we retained few equations that use these landmarks. We tested the male and combined sex equations on 36 males of unknown ethnicity exhumed from a military base in Guatemala, and found that the equations performed satisfactorily. We also evaluated the performance of equations by Steele ([1970] Personal Identification in Mass Disasters; Washington, DC: Smithsonian Institution) and Jacobs ([1992] Am J Phys Anthropol 89:333-345) on the Maya bones, and conclude that significant population variation in long bone proportions hinders their application in Central America. PMID- 12567378 TI - Variations in cortical material properties throughout the human dentate mandible. AB - Material properties and their variations in individual bone organs are important for understanding bone adaptation and quality at a tissue level, and are essential for accurate mechanical models. Yet material property variations have received little systematic study. Like all other material property studies in individual bone organs, studies of the human mandible are limited by a low number of both specimens and sampled regions. The aims of this study were to determine: 1) regional variability in mandibular material properties, 2) the effect of this variability on the modeling of mandibular function, and 3) the relationship of this variability to mandibular structure and function. We removed 31 samples on both facial and lingual cortices of 10 fresh adult dentate mandibles, measured cortical thickness and density, determined the directions of maximum stiffness with a pulse transmission ultrasonic technique, and calculated elastic properties from measured ultrasonic velocities. Results showed that each of these elastic properties in the dentate human mandible demonstrates unique regional variation. The direction of maximum stiffness was near parallel to the occlusal plane within the corpus. On the facial ramus, the direction of maximum stiffness was more vertically oriented. Several sites in the mandible did not show a consistent direction of maximum stiffness among specimens, although all specimens exhibited significant orthotropy. Mandibular cortical thickness varied significantly (P < 0.001) between sites, and decreased from 3.7 mm (SD = 0.9) anteriorly to 1.4 mm posteriorly (SD = 0.1). The cortical plate was also significantly thicker (P < 0.003) on the facial side than on the lingual side. Bone was 50-100% stiffer in the longitudinal direction (E(3), 20-30 GPa) than in the circumferential or tangential directions (E(2) or E(1); P < 0.001). The results suggest that material properties and directional variations have an important impact on mandibular mechanics. The accuracy of stresses calculated from strains and average material properties varies regionally, depending on variations in the direction of maximum stiffness and anisotropy. Stresses in some parts of the mandible can be more accurately calculated than in other regions. Limited evidence suggests that the orientations and anisotropies of cortical elastic properties correspond with features of cortical bone microstructure, although the relationship with functional stresses and strains is not clear. PMID- 12567379 TI - Patterns of joint size dimorphism in the elbow and knee of catarrhine primates. AB - Differences in body size between conspecific sexes may incur differences in the relative size and/or shape of load-bearing joints, potentially confounding our understanding of variation in the fossil record. More specifically, larger males may experience relatively greater limb joint stress levels than females, unless an increase in weight-related forces is compensated for by positive allometry of articular surface areas. This study examines variation in limb joint size dimorphism (JSD) among extant catarrhines to: 1) determine whether taxa exhibit JSD beyond that expected to simply maintain geometric similarity between sexes, and 2) test whether taxa differ in JSD (relative to body size dimorphism) with respect to differences in limb use and/or phylogeny. "Joint size" was quantified for the distal humerus and distal femur of 25 taxa. Analysis of variance was used to test for differences between sexes (in joint size ratios) and among taxa (in patterns of dimorphism). Multiple regression was used to examine differences in JSD among taxa after accounting for variation in body size dimorphism (BSD) and body size. Although degrees of humeral and femoral JSD tend to be the same within species, interspecific variation exists in the extent to which both joints are dimorphic relative to BSD. While most cercopithecoids exhibit relatively high degrees of JSD (i.e., positive allometry), nonhuman hominoids exhibit degrees of JSD closer to isometry. These results may reflect a fundamental distinction between cercopithecoids and hominoids in joint design. Overall, the results make more sense (from a mechanical standpoint) when adjustments to BSD are made to account for the larger effective female body mass associated with bearing offspring. In contrast to other hominoids, modern humans exhibit relatively high JSD in both the knee and elbow (despite lack of forelimb use in weight support). Estimates of BSD based on fossil limb bones will vary according to the extant analogue chosen for comparison. PMID- 12567380 TI - Peripartum cortisol levels and mother-infant interactions in Japanese macaques. AB - As evidence accumulates regarding the influence of hormones and stress-related conditions on maternal behavior, it becomes critical to better understand the relationship between physiological stress and the ability to cope with infants. Eight Japanese macaque females were observed 3 hr per week during the first 12 weeks after parturition; fecal samples were collected twice a week from each mother, starting 4 weeks before parturition and ending 4 weeks after parturition. Time spent in contact, maternal responsiveness, latency of response, and maternal rejection were measured and correlated with peripartum excreted cortisol and estradiol metabolite levels. Two indices of peripartum hormonal status were also tested against behavior: the postpartum stress index, and the postpartum cortisol/prepartum estradiol ratio (F/E). Postpartum cortisol levels showed a positive correlation with maternal rejection. The cortisol/estradiol ratio was positively correlated with rejection and latency of response, and negatively correlated with maternal responsiveness. Prepartum cortisol levels and the postpartum stress index did not correlate with any aspect of maternal behavior. Our findings suggest that hypothalamic-pituitary-adrenal (HPA) axis activity per se is not enough to predict the quality of interaction between mother and infant. Only when cortisol is high relative to estradiol could it be symptomatic of a possible negative feedback response involving stress, adrenal activity, and the ability of mothers to cope with the additional problems imposed by newborns. PMID- 12567381 TI - This commentary is based on the recently published article: Tantisira KG, Systrom DM, Ginns LC. An elevated breathing reserve index at the lactate threshold is a predictor of mortality in patients with cystic fibrosis awaiting lung transplantation. Am J Respir Crit Care Med 2002;165;1629-1633. PMID- 12567382 TI - Heads up! clear those airways! PMID- 12567383 TI - Negative expiratory pressure: a new tool for evaluating lung function in children? AB - The negative expiratory pressure technique (NEP) has been applied in adults with chronic obstructive pulmonary disease (COPD), demonstrating flow limitation in many of these patients. Because this technique does not require patient cooperation, it is of potential interest for application in the pediatric population. This study was performed to test the feasibility of NEP in children, and to further investigate it in children with asthma and cystic fibrosis (CF). We performed NEP (0.3-0.7 kPa) measurements in 14 healthy children (13.3 years, +/- 2.4), in 12 children with asthma (11.7 years, +/- 3.0), and in 17 children with CF (13.3 years, +/- 2.7). NEP-derived flow-volume loops were visually analyzed for flow limitation at tidal breathing. In addition, expiratory flow at 50% of tidal volume (TEF(50)) was measured. In healthy children, the intraclass coefficient of correlation was 77%, and intraindividual short- and long-term variability was 5.8% and 10.8%, respectively. In asthmatics, TEF(50) was lower compared with controls, and increased after inhalation of salbutamol. However, appropriate size-correction has still to be established. Measurement of TEF(50) using NEP is feasible in children. Despite good reproducibility in individual patients, the high intersubject variability may limit its usefulness as a clinical tool. In addition, the lack of flow limitation using NEP even in severely obstructed patients with CF warrants further investigation. PMID- 12567384 TI - Airway and tissue mechanics in anesthetized paralyzed children. AB - To estimate the mechanical properties of the airways and respiratory tissues, respiratory system impedance (Zrs) was measured with low-frequency forced oscillations in 26 anesthetized, paralyzed children (aged 3 months-10 years) undergoing surgical correction of congenital heart diseases. Zrs was determined from the signals of tracheal flow and pressure between 0.4-12 Hz before surgery at zero mean transrespiratory pressure. The pulmonary (Z(L)) and chest wall (Z(W)) components of Zrs were also determined in 5 children by measuring esophageal pressure. A model containing frequency-independent resistance (R) and inertance (I), and coefficients of tissue-damping (G) and elastance (H), was fitted to the Zrs, Z(L), and Z(W) spectra. The total respiratory parameters normalized to body weights were 82.2 +/- 8.5 (SE) hPa x sec x l(-1) x kg, 0.152 +/- 0.05 hPa x sec(2) x l(-1) x kg, 293.8 +/- 20.0 hPa. l(-1) x kg, and 1,583 +/- 65.5 hPa x l(-1) x kg, for R, I, G, and H, respectively. The measurements of Z(L) and Z(W) revealed the dominance of the lungs in R (91 +/- 4.3%) and I (109 +/- 16%), and the major contribution of the lung parenchyma to G (61 +/- 7.3%) and H (66 +/- 7.4%) of the total respiratory system. It is concluded that anesthesia paralysis provides an ideal condition for the measurement of low-frequency forced oscillatory impedance and its partitioning into airway and tissue components in mechanically ventilated children. The separation of pulmonary and chest wall mechanics demonstrates that airway properties can be estimated appropriately from Zrs data, while the chest wall may damp the changes in parenchymal properties. PMID- 12567385 TI - Spirometric function in children of Mexico City compared to Mexican-American children. AB - We set out to describe the pattern of lung function growth in Mexican students from 8-20 years of age, using internationally accepted equipment and methodology, and to compare it to values reported for Mexican-American children. Out of a total of 6,803 students from primary school to high school studied cross sectionally in the Mexico City metropolitan area, we selected 4,009 asymptomatic, nonobese, nonsmoker subjects to generate spirometric prediction equations. We describe regression equations for the main spirometric variables (log transformed) based on age, height, and weight, and separated for males and females. Spirometric function in the population studied was above that predicted for European (Quanjer et al. [1987] Pediatr Pulmonol 19:135-142) or Mexican American children, for the same age, height, and gender. On average, forced expiratory volume in 1 sec (FEV(1)) in Mexican children was 9.5% above that of Europeans (Quanjer et al. [1987] Pediatr Pulmonol 19:135-142), 14% and 5% above Hispanics reported by (Coultas et al. [1988] Am Rev Respir Dis 138:1386-1392) and (Hsu et al. [1979] J Pediatr 95:14-23), respectively, and 5% above Mexican Americans from the third National Health and Nutrition Examination Survey study. Similarly, FVC was 8%, 14%, 8%, and 5.6% above the figures predicted by the same authors. The largest errors of prediction of foreign equations occurred in extremely tall or short subjects, and therefore a single proportional adjustment is unfeasible. PMID- 12567386 TI - Viral respiratory infections in young children attending day care in urban Northeast Brazil. AB - A cohort of children attending a day care center in Salvador (Bahia, Brazil) was studied prospectively to determine the incidence of viral respiratory infectious episodes and to identify the viruses associated with them. Two hundred seventy one nasopharyngeal samples were collected over a 1-year period for examination, using indirect immunofluorescence with monoclonal antibodies against adenovirus, influenza A and B, parainfluenzae 1-3, and respiratory syncytial virus, and reverse transcriptase-polymerase chain reaction for picornavirus. Examination yielded positive results in 116 samples (42.8%). Rhinovirus was identified alone in 56 samples (48.3%) and was observed along with other viruses in 11 additional samples. Incidence density of viral respiratory infectious episodes was 7.66 episodes/1,000 child-days. PMID- 12567387 TI - Influence of particle size on lung deposition and pharmacokinetics of beclomethasone dipropionate in children. AB - We set out to evaluate lung deposition, systemic availability, and basic pharmacokinetic parameters of beclomethasone dipropionate (BDP) in children with chronic asthma. Plasma levels of BDP, 17 and 21 beclomethasone monopropionate (17 BMP and 21-BMP), and beclomethasone were measured after an intravenous infusion of 60 microg BDP and after inhalation of A) 100 microg HFA-BDP, B) 200 microg HFA BDP, C) 200 microg HFA-BDP after ingestion of charcoal to block gastrointestinal (GI) absorption of drug, and D) 400 microg CFC-BDP. A breath-actuated pMDI (Autohaler) was used for HFA inhalations, and a pMDI with a large volume spacer (Volumatic) for CFC inhalations. Treatments A-D were given in a randomized, cross over design. Fourteen patients aged 10-14 years completed all 5 study days. The mean systemic bioavailabilities in percent of dose leaving the canister valve (ex valve) were 70% (100 HFA), 74% (200 HFA), 60% (200 HFA + charcoal), and 27% (400 microg CFC). After HFA treatment, 82% of the systemically available dose was absorbed through the lungs, and 18% from the gastrointestinal tract. The estimated bioavailability of BDP from the GI tract was 68%. BDP was metabolized to 17-BMP within minutes. Mean steady-state volume of distribution of 17-BMP was 84 L, and the mean terminal half-life (T((1/2))) after the four inhalations was 2.7 hr (range, 2.2-3.7 hr). Mean T((1/2)) and clearance after i.v. administration were 1.7 hr and 0.9 L/min, respectively. The HFA Autohaler delivers approximately three times as much BDP to the intrapulmonary airways as a CFC-pMDI with a large volume spacer. PMID- 12567388 TI - Forced expiratory parameters in healthy preschool children (3-6 years of age). AB - In a group of 173 healthy preschool children 3-6 years of age (body height, 90 130 cm; 102 boys and 71 girls) out of total 279 children examined, maximum expiratory flow-volume (MEFV) curves were recorded in cross-sectional measurements. The majority (62%) of preschool children were able to generate an MEFV curve as correctly as older children. From the curves, maximum expiratory flows at 25%, 50%, and 75 % of vital capacity (MEF(25), MEF(50), and MEF(75)), peak expiratory flow (PEF), forced expiratory volume in 1 sec (FEV(1)), forced vital capacity (FVC), and area delineated by MEFV curve (A(ex)) were obtained. The purpose of the study was to establish reference values of forced expiratory parameters in preschool children suitable for assessment of lung function abnormalities in respiratory preschool children. The values of the studied parameters increased nonlinearly and correlated significantly with body height (P < 0.0001); the correlation was much lower with age. A simple power regression equation was calculated for the relationship between each parameter and body height. A best-fit regression equation relating functional parameters and body height was a power function. Based on the obtained regression equations with upper and lower limits, we prepared tables listing reference values of forced expiratory parameters in healthy Caucasian preschool children, against which patients can be compared. No statistically significant gender differences were observed for MEF(25), MEF(50), MEF(75), PEF, FEV(1), FVC, and A(ex) by extrapolation. The reference values were close to those obtained in our older children. A decline of the ratios PEF/FVC, FEV(1)/FVC and MEF/FVC with increasing body height suggested more patent airways in younger and smaller preschool children. PMID- 12567389 TI - Chest physiotherapy in infants with cystic fibrosis: to tip or not? A five-year study. AB - There is controversy about the need for postural drainage physiotherapy in asymptomatic infants with cystic fibrosis (CF). We aimed to compare the effectiveness of standard postural drainage chest physiotherapy (SPT) with a modified physiotherapy regimen without head-down tilt (MPT) in young infants with CF. Twenty newly diagnosed infants with CF (mean age, 2.1 months; range, 1-4) were randomized to SPT or MPT. Parents kept a detailed symptom and treatment diary for the following 12 months. Serial chest radiographs, taken at diagnosis, 12 months, 2(1/2) years, and 5 years after diagnosis, were assessed using the Brasfield score. Pulmonary function tests were compared between groups after 5 years. Of the 20 infants, 16 (80%) completed the review at 12 months, and 14 (70%) at 2(1/2) and 5 years. Patients receiving SPT had more days with upper respiratory tract symptoms than those on MPT (70 +/- 32.8 vs. 37 +/- 24.9 days; P = 0.04) and required longer courses of antibiotics (23 +/- 28.5 vs. 14 +/- 11.2 days; P = 0.05). Chest x-ray scores were similar at diagnosis but were worse at 2(1/2) years for those receiving SPT (P = 0.03). Forced vital capacity and forced expired volume in 1 sec (FEV(1)) at 5-6 years was lower for SPT than for MPT (P < 0.05). In conclusion, MPT was associated with fewer respiratory complications than SPT in infants with CF. PMID- 12567390 TI - Impulse oscillometry: a measure for airway obstruction. AB - The impulse oscillometry system (IOS) was introduced as a new technique to assess airflow obstruction in patients who are not able to perform forced breathing maneuvers, e.g., subjects with cerebral palsy or severe mental retardation, and young children. This study evaluates the sensitivity and specificity of IOS parameters to quantify changes in airflow obstruction in comparison with forced expiratory volume in the first second (FEV(1)) and peak expiratory flow (PEF) measurements. Measurements of FEV(1), PEF, and resistance (R) and reactance (X) at frequencies of 5-35 Hz were performed in 19 children with asthma before, during, and after methacholine challenge and subsequent bronchodilatation. All parameters changed significantly during tests. Values of R5 and R10 correlated with FEV(1) (r = -0.71 and -0.73, respectively, P < 0.001), as did values of X5 and X10 (r = 0.52 and 0.57, respectively, P < 0.01). Changes in R preceded changes in PEF and FEV(1) during methacholine challenge. The area under the receiver operating characteristic (ROC) curve to predict a 15% fall in FEV(1) showed better sensitivity and specificity for R5 (area under the curve, 0.85) compared to PEF (0.79) or R10 (0.73). We conclude that IOS parameters can be easily used as an indirect measure of airflow obstruction. This might be helpful in patients who are not able to perform forced breathing maneuvers. In individual subjects, R values measured at 5 Hz showed to be superior to PEF measurements in the detection of a 15% fall in FEV(1). PMID- 12567391 TI - Inspiratory flow rate through a dry powder inhaler (Clickhaler) in children with asthma. AB - Dry powder inhalers (DPIs) are increasingly being used to deliver drugs for the treatment of asthma. Both the aerosolization and delivery of the drug from a DPI to the lung are dependent on an adequate inspiratory effort from the patient, and it is well-known that the air flow achieved early in the inspiratory profile is important in determining particle size distribution from the inhaler. The present study assessed the peak inspiratory flow (PIF) generated through the Clickhaler DPI, and the early inspiratory flow at 150 mL of inspired volume (IF(150)), in asthmatic children. These measurements were made in a well-controlled setting, and two attempts were recorded to establish maximum achievement. Results were obtained from 57 children aged 6-17 years, showing a (mean +/- SD) best PIF of 60.5 +/- 18.7 L/min (range, 26.8-97.0). The mean PIF overall was 54.2 +/- 20.8 L/min (7.9-97.0). For children aged 6-8 years, the mean best PIF was 46.5 +/- 14.6 L/min (26.8-71.1); for those aged 9 years or more, it was >65 L/min (30.3 97.0). PIF values were unrelated to % predicted FEV(1) measurements. Best IF(150) (mean +/- SD) was 42.9 +/- 13.6 L/min (23.1-66.6) in children aged 6-8 years, and >55 L/min (28.0-86.4) for the older children, showing that high flow rates were achieved early in the inspiratory profile. These data indicate that children with stable asthma can generate adequate inspiratory flow rates to operate the Clickhaler effectively. PMID- 12567392 TI - Case of congenital bronchial atresia detected by fetal ultrasound. AB - Congenital bronchial atresia is an anomaly characterized by a mucocele associated with a blindly terminating segmental or lobar bronchus. We report on a case of congenital atresia of the left superior segmental bronchus (B6) followed by fetal ultrasonography, which showed a large simple cyst with a surrounding small echogenic area. A large cyst and persistent lung fluid in the affected segment were noted in the early neonatal period. PMID- 12567393 TI - Chylothorax following apparently spontaneous central venous thrombosis in a patient with septic shock. AB - Within the pediatric age group, chylothorax is rare and has been reported almost exclusively in the setting of thoracic surgical procedures or central venous hypertension secondary to central venous catheter thrombosis. We report on the development of central venous thrombosis and chylothorax in the absence of the usual risk factors in a patient with septic shock, and we expand on the role that procoagulant states, such as those induced by sepsis, might play in the development of this complication. This case reminds the practitioner that central venous thromboses and their complications may occur in the absence of the usually reported risk factors and must therefore still be considered when other clinical events suggest their presence. PMID- 12567394 TI - Chronic interstitial lung disease with lung fibrosis in a girl: uncommon sequelae of Epstein-Barr virus infection. AB - A 12-month-old immunocompetent girl presented with tachypnea, inspiratory crackles, mild hypoxemia, and failure to thrive after an acute Epstein-Barr virus (EBV) infection. The course of acute EBV infection was demonstrated by viral load measurement in plasma and peripheral blood mononuclear cells (PBMC) by using real time polymerase chain reaction (PCR). EBV DNA was further detected by PCR in bronchoalveolar lavage (BAL) fluid and from a lung-tissue specimen obtained by open-lung biopsy, which indicates the pulmonary involvement of active EBV infection. Histology revealed an uncharacteristic interstitial infiltration and fibrosis. Following topic and systemic treatment with corticosteroids, the child became asymptomatic and showed normal weight gain as well as mental and physical development. Pulmonary parenchymal involvement during the course of primary EBV infection may result in interstitial lung disease and fibrosis not only in immunodeficient, but in immunocompetent children as well. Treatment with combined inhaled and oral steroids seems to be a treatment option in these patients. PMID- 12567395 TI - Human uterine vascular structures in normal and diseased states. AB - Menstrual bleeding disorders are one of the most common problems presenting to the gynaecologist. There is increasing evidence that alterations in human uterine vascular structures are associated with spontaneous and sex steroid-induced changes in menstrual bleeding patterns. This article will discuss the normal anatomy and physiology of uterine vascular growth, breakdown, and repair and will indicate where altered anatomy and function may contribute to menstrual disorder. In particular, impact of low-dose progestogens on the endometrial vasculature and endometrial vascular fragility will be discussed. Disturbances of myometrial vascular growth and remodeling, including fibroids, adenomyosis, implantation, and arterio-venous malformations will also be addressed. PMID- 12567396 TI - Modification of uterine vasculature during pregnancy in macaques. AB - Embryonic development in macaques includes extensive modification of the uterine vasculature by fetal trophoblast cells. Soon after the onset of blastocyst attachment to the endometrium, syncytial trophoblast cells intrude between endometrial epithelial cells, resulting in focal epithelium loss. Trophoblast cells continue to move into the endometrial stroma and encounter superficial uterine capillaries. These capillaries are penetrated by trophoblast, which permits maternal blood to leave the maternal circulation and enter lacunae formed within the mass of trophoblast cells. Cytotrophoblast cells enter the uterine vessels and attach to the endothelium via cell adhesion molecules prior to migration into confluent spiral arterioles, against the flow of blood. As intra arterial cytotrophoblast cells migrate, they displace adjacent endothelium, produce matrix metalloproteinases, traverse the tunica intima, and reside in the tunica media as intramural trophoblast. Intramural trophoblast cells disrupt the tunica media and become surrounded by an extensive extracellular matrix. In areas proximal to the placenta, the entire circumferences of spiral arteries are modified in this way. In the same arteries, distal to the placenta and farther "upstream," trophoblast-mediated changes to the arterial wall are less extensive. Uterine veins are modified by trophoblast only in the area immediately next to the trophoblast shell, with no trophoblast migration. The functional consequence of this trophoblast activity may be to ensure an adequate flow of maternal blood to the placenta, thus enhancing the survival of the fetus. PMID- 12567397 TI - Development of the utero-placental circulation: the role of carbon monoxide and nitric oxide in trophoblast invasion and spiral artery transformation. AB - It is now well known that in order to establish human hemochorial placentation and to provide a progressive increase in blood supply to the growing fetus, the uterine spiral arteries must undergo considerable alterations. This physiological modification is thought to be brought about by the interaction of invasive cytotrophoblast with the spiral artery vessel wall. Despite intensive research our understanding of the mechanisms that control human trophoblast invasion in normal, let alone abnormal pregnancy, are sill poorly understood. This is partly due to difficulties in obtaining "true" placental bed biopsies and most investigators have relied on in vitro models of trophoblast invasion. This article describes the morphological changes that occur within the placental bed throughout human pregnancy along with a review of the various studies which have attempted to sample the placental bed. Thereafter, follows a review of the evidence that invasive trophoblast can release the vasoactive agents nitric oxide and/or carbon monoxide which, in turn, could contribute to early physiological changes in spiral arteries prior to destruction of the smooth muscle within the vessel wall. Current evidence supports the idea that trophoblast-derived carbon monoxide may contribute to spiral artery modification. In contrast there is no evidence for a similar role by nitric oxide. PMID- 12567398 TI - Endometrial arteriogenesis: vascular smooth muscle cell proliferation and differentiation during the menstrual cycle and changes associated with endometrial bleeding disorders. AB - The cyclical regrowth of the human endometrial vasculature every 28 days following menstruation provides an excellent opportunity for studying arteriogenesis in a human tissue. Despite this, very little has been published to date about temporal or spatial changes in endometrial arteriolar structure or function. Immunohistochemical studies using vascular smooth muscle cell (VSMC) differentiation markers have identified spatially organised differences in VSMC phenotype. However, these have not significantly increased our understanding of how or when arteriogenesis occurs. Endometrial VSMC proliferation continues at a steady rate throughout the cycle, increasing in the specialised spiral arterioles in the mid and late secretory phases. Although estrogen and progesterone play a dominant role in regulating endometrial growth and regression, their direct involvement in arteriogenesis is less clear. Estrogen and progesterone receptors have been reported in endometrial VSMC, although heterogeneity of expression occurs. The angiopoietin/Tie axis has been investigated in endometrium, although the results of 3 studies published to date are contradictory, making conclusions about a role in arteriogenesis problematic. Abnormalities of arteriogenesis may play a role in disorders such as menorrhagia and breakthrough bleeding in contraceptive and hormone replacement therapy users. There is evidence that VSMC proliferation is reduced in spiral arterioles of women with menorrhagia, along with reduced pulsatility index of the uterine artery as shown by Doppler ultrasound. In women taking progestin-only contraception, endometrial perivascular alpha smooth muscle actin is reduced in those who suffer from breakthrough bleeding compared to those that don't. Considerable work is still required to elucidate when in the cycle, where in the tissue, and what mechanisms regulate endometrial arteriogenesis. PMID- 12567399 TI - Granulated metrial gland cells in the murine uterus: localization, kinetics, and the functional role in angiogenesis during pregnancy. AB - Granulated metrial gland (GMG) cells are a major immune cell population in the murine pregnant uterus, and contribute to the maintenance of pregnancy by functioning as uterus-specific natural killer (NK) cells. In order to reveal their kinetics, activation, and functional roles in pregnancy, we conducted quantitative and immunohistochemical analyses in normal and immuno-modulator treated mice. Under a light microscope, GMG cells were identified by red cytoplasmic granules in periodic-acid-Schiff (PAS)-stained sections. They progressively increased in number and size with the peak at day 12-14 of pregnancy in the decidua and metrial gland. New vessel formation was most prominent around day 8, and the total vascular area reached the peak at day 13. GMG cells were often located near the blood vessels, and expressed vascular endothelial growth factor (VEGF), suggesting their possible inducing role in angiogenesis during the development of decidua/metrial gland. While blood vessels in the non-pregnant uterus were negative for vascular cell adhesion molecule (VCAM)-1, those in the pregnant one were positive. Treatment with neutralizing antibody against VCAM-1, however, did not decrease the number of GMG cells. On the other hand, mitosis of GMG cells was frequently observed. These data suggest that the increment of GMG cells during pregnancy may largely result from local proliferation in the uterus rather than an increased influx of precursor cells. Although we attempted to induce in vivo activation of GMG cells by administration of interleukin-12 (IL-12) or alpha-galactosylceramide, a potent activator for natural killer-T (NK-T) cells, the number of GMG cells did not appreciably increase. The present study has demonstrated that GMG cells locally proliferate in the pregnant uterus, not being related to VCAM-1 expression by the uterine vasculature or systemic activation of NK cells and NK-T cells, and seem to be involved in angiogenesis in the pregnant uterus through VEGF production. PMID- 12567400 TI - Expression of adhesion molecules in the lower uterine segment during term and preterm parturition. AB - Term and preterm cervical ripening and dilatation have similarities with an inflammatory reaction. Since cell adhesion molecules are involved in this process, investigations on the expression of intercellular adhesion molecule-1, endothelial leukocyte adhesion molecule-1, vascular cell adhesion molecule-1, and platelet endothelial cell adhesion molecule in the lower uterine segment and in vitro experiments on human umbilical vein endothelial cells were performed. In addition, current reports on expression of endothelial adhesion molecules by the uterine cervix were summarized. Cell adhesion molecule expression by lower uterine segment and uterine cervix in term and preterm parturition was measured using immunohistochemistry, enzyme immunoassay, and Northern blot analysis. Regulation of adhesion molecule expression was evaluated in vitro by indirect immunofluorescence and flow cytometry using human umbilical vein endothelial cells. Investigations in term parturition revealed that intercellular adhesion molecule-1, endothelial leukocyte adhesion molecule-1, and vascular cell adhesion molecule-1 expression increases during parturition. In preterm labor, the expression of endothelial leukocyte adhesion molecule-1 and intercellular adhesion molecule-1 in the lower uterine segment increased. Expression of platelet endothelial cell adhesion molecule did not change in term and preterm parturition. Expression of adhesion molecules was localized mainly on lower uterine segment vascular endothelial cells and to a smaller extent on leukocytes. In vitro experiments showed that expression of adhesion molecules by human umbilical vein endothelial cells can be stimulated by tumor necrosis factor alpha, 17beta-estradiol, prostaglandin E(2), and the antigestagen onapristone. Progesterone exerted no stimulatory effect. Cervical ripening and dilatation during term and preterm parturition are associated with an increased expression of endothelial cell adhesion molecules by lower uterine segment and uterine cervix. The expression can be modulated by pro-inflammatory cytokines, sex hormones, and prostaglandin E(2). Mechanisms controlling the extravasation of leukocytes may play a fundamental role in term and preterm parturition. PMID- 12567401 TI - Stromal vascularization in the endometrium during adenomyosis. AB - In adenomyosis, the two major frequent symptoms are hypermenorrhea and dysmenorrhea. The incidence of the former is said to be high, ranging from 36 to 70%. It has been found that vascular distribution of the endometrium in adenomyosis is diversely different from fertile patients without adenomyosis. For example, hysteroscopy revealed that approximately half of the patients have abnormal vascularization. In these patients, vascular distribution was generally irregular, and vessels were thick, dilated, and/or reticular in some patients. Moreover, morphometric analysis of the endometrium revealed that in the fertile women, the mean surface area, total surface area, and total number of capillaries, all increased significantly in the secretory phase compared to the proliferative phase. In contrast, the above parameters increased in the adenomyosis group in both the proliferative phase and secretory phase compared to the fertile women. In particular, the total surface area of capillaries per mm(2) markedly rose, by 11.6 times, compared to that of the proliferative phase in the fertile women. These findings suggest that regulatory factors involved in the vascular proliferation are diversely exaggerated, and that the abnormal vascularization of the endometrium is closely related with hypermenorrhea. PMID- 12567402 TI - Synaptic process in the rat (Rattus norvegicus): Influence of methodology on results. AB - Synaptonemal complex (SC) analysis is a widely used method for assessing the effects of genotoxic agents in germ cells. Although the evolution of the SCs and their related annexed structures, such as nucleoli, has been well established, sometimes it is difficult to assess whether the abnormal features observed correspond to genotoxic effects or to an artefact related to the method used to obtain the SC preparations. In this article, we describe a new method of obtaining SC preparations for electron microscopy, as well as the results of a study of the first meiotic prophase in oocytes and spermatocytes of the rat (Rattus norvegicus Sprague Dawley) in which we analysed how the methodology used can influence the results. Besides important sex-specific differences, mainly during desynapsis (diplotene), a relationship between several bivalents and nucleolar structures, that in some cases could disturb the synaptic process, was observed in oocytes. At the same time, the characteristic SC fragmentation in oocytes was verified, but this fragmentation, in addition to a sex-specific component, was influenced by the method itself. By reducing to a minimum the artefacts produced by the method, it is possible to optimise the analysis of SCs as a method of testing genotoxic effects in the germ line. PMID- 12567403 TI - Use of the confocal laser scanning microscope in studies on the developmental biology of marine crustaceans. AB - Confocal Laser Scanning Microscope techniques have been applied to study the developmental biology of marine copepods and decapod larvae. The lipophylic probes DiI and DiOC(6) were used to study both the external and internal morphology of these crustaceans, whereas the same DiOC(6) and the specific nuclear probe Hoechst 33342 were used to study embryonic development of copepods in vivo. To distinguish viable from non-viable copepod embryos, the vital dye dichlorodihydrofluorescein diacetate (H(2)DCFDA) was used. Major advantages and difficulties in the use of these non-invasive techniques in studies of the reproductive biology of marine crustaceans are discussed. PMID- 12567404 TI - A simple method to observe organic solvent drops with a standard optical microscope. PMID- 12567405 TI - Routine cytogenetic and FISH studies for 17p11/15q11 duplications and subtelomeric rearrangement studies in children with autism spectrum disorders. AB - To assess the frequency of cytogenetic abnormalities in children with autism spectrum disorders (ASDs), routine G-banded cytogenetic analyses and FISH studies to rule out 15q11.2 and 17p11.2 duplications were performed on 49 children with ASDs. Blood samples were further studied using a complete set of subtelomeric FISH probes. Routine chromosome study showed that one child had a small duplication of chromosome 5: 46,XY,dup(5)(p?14.2p?15.1). Another child had an interstitial duplication of the Prader-Willi and Angelman syndrome critical region of chromosome 15, detected by FISH analysis. The detection of these two cases underscores the importance of obtaining routine chromosome and 15q11-q13 FISH analyses in children with ASDs. No instance of 17p11.2 duplication was observed. Subtelomeric analysis did not reveal abnormalities in any of the subjects. PMID- 12567406 TI - Previously undescribed nonsense mutation in SHH caused autosomal dominant holoprosencephaly with wide intrafamilial variability. AB - Holoprosencephaly (HPE) is the most common developmental defect of the forebrain and midface in humans, with a frequency of 1/16,000 live births. Different genes are implicated in the pathogenesis of HPE; these include SHH, ZIC2, SIX3, TGIF, and human DKK1. We describe here a family with recurrence of autosomal dominant HPE in different members showing a wide clinical variability. The mother presents a single central maxillary incisor and mild hypotelorism as signs of the diseases, while three of her sons were affected by HPE. By direct sequencing and restriction analysis of exon 2 of the SHH gene, we have identified a previously undescribed nonsense mutation at codon 128 (W128X). The identification of this mutation allowed us to give a prenatal diagnosis in this family and confirms a wide intrafamilial variability in the phenotypic spectrum. PMID- 12567407 TI - Genetic heterogeneity in inherited spastic paraplegia associated with epilepsy. AB - We have recently mapped a new rare form of spastic paraplegia complicated by bilateral cataracts, gastroesophageal reflux with persistent vomiting, and amyotrophy to chromosome 10q23.3-q24.2. This locus, named SPG9, is located in an interval spanning about 12 cM of genomic DNA, between markers D10S536 and D10S603, where different neurological disorders have been mapped. In particular, a gene for partial epilepsy has been assigned to a 3 cM interval between markers D10S185 and D10S577, which is completely included in the SPG9 critical region. A few families affected with spastic paraplegia and epilepsy have been reported; in the present study, we tested a pedigree with concurrence of spastic paraplegia, epilepsy, and mental retardation inherited as an autosomal dominant trait, using markers located in the SPG9 interval. Haplotype reconstruction excluded the linkage to 10q23.3-q24.2. In addition, the seven different loci so far reported to be associated with autosomal dominant pure forms of spastic paraplegia have been tested and excluded by linkage analysis and haplotype reconstruction, including SPG4 on chromosome 2p22-p21, where a familial form of spastic paraplegia associated with dementia and epilepsy has been mapped. These data confirm genetic heterogeneity in familial spastic paraplegia with epilepsy and suggest a specific locus for the family here analyzed. PMID- 12567408 TI - Molecular characterization of a 12q22-q24 deletion associated with congenital deafness: confirmation and refinement of the DFNA25 locus. AB - The DFNA25 locus for autosomal dominant nonsyndromic hereditary hearing loss has been mapped to 12q21-q24 by linkage analysis. A de novo deletion in a six-year old boy with congenital hearing loss as well as mental and motor retardation now provides independent confirmation of this genetic localization and narrows the critical interval to 13 cM in the 12q22-q24.1 region. Mapping of the deletion was performed using fluorescence in situ hybridization (FISH) analysis with region specific yeast artificial chromosome (YAC) clones. Ten YACs 929_e_4, 959_c_3, 746_h_7, 817_h_10, 886_a_6, 916_h_9, 969_c_12, 747_e_2, 812_h_12, and 959_f_8 were absent from one chromosome 12 from the patient. Molecular analyses of eight polymorphic markers helped to narrow down the breakpoints and demonstrated that the derivative chromosome 12 is of paternal origin. Several known genes including ATP2A2, UBE3B, and VR-OAC that map in the 12q22-q24.1 region are included in the deletion. These results provide evidence that haploinsufficiency for a gene or genes in 12q22-q24.1 is associated with autosomal dominant deafness. PMID- 12567409 TI - Clinical and genetic aspects of trigonocephaly: a study of 25 cases. AB - We reviewed 25 patients ascertained through the finding of trigonocephaly/metopic synostosis as a prominent manifestation. In 16 patients, trigonocephaly/metopic synostosis was the only significant finding (64%); 2 patients had metopic/sagittal synostosis (8%) and in 7 patients the trigonocephaly was part of a syndrome (28%). Among the nonsyndromic cases, 12 were males and 6 were females and the sex ratio was 2 M:1 F. Only one patient with isolated trigonocephaly had an affected parent (5.6%). All nonsyndromic patients had normal psychomotor development. In 2 patients with isolated metopic/sagittal synostosis, FGFR2 and FGFR3 mutations were studied and none were detected. Among the syndromic cases, two had Jacobsen syndrome associated with deletion of chromosome 11q 23 (28.5%). Of the remaining five syndromic cases, different conditions were found including Say-Meyer syndrome, multiple congenital anomalies and bilateral retinoblastoma with no detectable deletion in chromosome 13q14.2 by G-banding chromosomal analysis and FISH, I-cell disease, a new acrocraniofacial dysostosis syndrome, and Opitz C trigonocephaly syndrome. The last two patients were studied for cryptic chromosomal rearrangements, with SKY and subtelomeric FISH probes. Also FGFR2 and FGFR3 mutations were studied in two syndromic cases, but none were found. This study demonstrates that the majority of cases with nonsyndromic trigonocephaly are sporadic and benign, apart from the associated cosmetic implications. Syndromic trigonocephaly cases are causally heterogeneous and associated with chromosomal as well as single gene disorders. An investigation to delineate the underlying cause of trigonocephaly is indicated because of its important implications on medical management for the patient and the reproductive plans for the family. PMID- 12567410 TI - Broad phenotypic spectrum caused by an identical heterozygous CDMP-1 mutation in three unrelated families. AB - CDMP-1, a cartilage-specific member of the TGFss superfamily of secreted signaling molecules, plays a key role in chondrogenesis, growth and patterning of the developing vertebrate skeleton. Homozygous CDMP-1 mutations cause Hunter Thompson and Grebe types of acromesomelic chondrodysplasia and DuPan syndrome in humans, as well as brachypodism in mice, while heterozygous mutations cause brachydactyly type C (BDC). We present clinical and radiographic data from three unrelated families in which 12 members share the same heterozygous CDMP-1 mutation, an insertion (insG206), resulting in a frameshift predicted to cause functional haploinsufficiency. Although eight mutation carriers display BDC, four have normal hands and feet, confirming nonpenetrance of BDC with CDMP-1 mutations. In addition, several carriers have other skeletal abnormalities, including severe bilateral vertical talus (in two), developmental hip dysplasia (in one), and short stature (in two, who are otherwise unaffected). Premature vertebral end-plate disease was observed in four mutation carriers and was associated with spondylolysis and spondylolisthesis in three of these. Axial skeletal involvement has not been previously reported in association with CDMP-1 mutations. This finding is consistent with CDMP-1 expression in human hypertrophic chondrocytes, which are present in the ring epiphyses of vertebral end plates. Phenotypic variation in BDC has previously been attributed either to locus heterogeneity or to the varied functional effects of different CDMP-1 mutations. The remarkable range of phenotypes caused by this identical CDMP-1 mutation in these families emphasizes the crucial role of genetic background, stochastic variation and/or environmental factors in modifying the observed phenotype. Our findings illustrate that nonpenetrance for the typical features of BDC can be appreciable and that atypical skeletal features that have been reported in some patients with BDC (i.e., clubfoot, short stature, spondylolysis) may also result from CDMP-1 mutation. PMID- 12567411 TI - FOXL2-mutations in blepharophimosis-ptosis-epicanthus inversus syndrome (BPES); challenges for genetic counseling in female patients. AB - Mutations in the forkhead transcription factor gene 2 (FOXL2) were recently reported to cause blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) types I and II. Evidence was provided that BPES type I (eyelid abnormalities and female infertility) is caused by mutations resulting in a truncated FOXL2 protein. In contrast, mutant FOXL2 proteins, either with inserted aminoacids in the forkhead domain or polyalanine tract, or with novel aminoacids at the carboxyl end, were found in BPES type II, in which fertility is generally normal. We report a 32-year-old female patient with sporadic BPES and a history of menstrual cycle irregularities and periods of secondary amenorrhoea. A heterozygous frameshift mutation (c959-960insG) was found in the FOXL2 gene, resulting in a predicted FOXL2 protein with 212 novel aminoacids in the carboxyl end, suggesting BPES type II despite menstrual irregularities. The clinical presentations of our patient and of three female patients with BPES type II in the report of De Baere et al. [2001: Hum Mol Genet 10:1591-1600.] indicate phenotypic overlap between BPES type I and II. These observations do not support a clear-cut prediction of female fertility based on the FOXL2 molecular defect. As a consequence, FOXL2 mutation testing in female patients of child-bearing age with BPES should be handled with caution, and a two-step genetic counseling approach, including an initial pre-test information session, is proposed. PMID- 12567412 TI - Spondyloepimetaphyseal dysplasia with joint laxity leptodactylic form: clinical course and phenotypic variations in four patients. AB - We describe a 5-year-old boy and a 33-year-old woman with spondyloepimetaphyseal dysplasia with joint laxity leptodactylic form (spondyloepimetaphyseal dysplasia with multiple dislocations) (MIM 6003546), and two 12-year-old girls with the disorder who were previously reported as examples of a variant of sponatrime dysplasia. Their clinical manifestations included midface hypoplasia, micromelic short stature, and generalized joint laxity that caused multiple joint problems, including thoracolumbar scoliosis, hip subluxation, progressive genu valgum with knee and patellar subluxation, elbow subluxation, and malalignment of the wrist. Laryngotracheomalacia was present in two individuals, and myopathy was noted in one. The radiological findings in the four individuals included mild platyspondyly most conspicuous in infancy, narrow interpediculate distances of the lumbar spine evident in infancy, retarded epiphyseal ossification that evolved to epiphyseal dysplasia and later to degenerative joint disease, metaphyseal irregularities and striations present in early childhood, and leptodactylic appearance (slender short tubular bones) of the hand. PMID- 12567413 TI - Early use of clinical BRCA1/2 testing: associations with race and breast cancer risk. AB - When BRCA1/2 testing became commercially available in 1996, many U.S. experts voiced concern about the potential for indiscriminate use of testing among low risk women. Supporting this concern, several early surveys of interest in genetic testing suggested that genetic testing for cancer susceptibility might appeal most to individuals at low risk of carrying a mutation. To identify factors associated with early use of clinical BRCA1/2 testing, a case-control study was conducted at a large academic health system in the metropolitan Philadelphia region. A total of 167 women underwent genetic counseling for clinical BRCA1/2 testing between 1996 and 1997 (cases) compared with 138 women who were seen in faculty general internal medicine practices over the same period (controls). In this study we measured the risk factors for breast cancer, the risk factors for carrying a BRCA1/2 mutation, and sociodemographic characteristics. Use of BRCA1/2 counseling between 1996 and 1997 was positively associated with family but no personal history of breast cancer (odds ratio (OR), 22.4; 95% confidence interval (CI), 9.3-54.3); family and personal history of breast cancer (OR, 150.3; 95% CI, 24.1-939.6); being Caucasian and non-Jewish (OR, 4.1; 95% CI, 1.3-13.5); being Caucasian and Jewish (OR, 8.8; 95% CI, 2.2-35.5); and being married (OR, 3.2; 95% CI, 1.6-6.3). Use of BRCA1/2 counseling was inversely associated with increasing age (OR, 0.07; 95% CI, 0.02-0.28 for >60 compared to <50). As suggested by the association with family history, use of counseling was associated with having a higher predicted risk of breast cancer and a higher predicted risk of carrying a BRCA1 mutation (P < 0.0001). Women who sought clinical BRCA1/2 testing in the year after it became commercially available were not the "worried well," but women at significantly increased risk of carrying a mutation. However, even after adjusting for breast cancer risk, there was a substantial racial disparity in use of BRCA1/2 testing. These findings suggest that ensuring equal access to testing for high-risk individuals irrespective of race may be as important for the future of predictive genetic testing as restricting the use of testing among low-risk individuals. PMID- 12567414 TI - Somatic instability of the androgen receptor CAG repeat in a normal female. AB - The polyglutamine repeat disorders represent a family of degenerative neurological diseases which are characterized by expansions of tandemly repeated CAG repeats. Investigations have demonstrated that in Huntington disease, dentatorubral pallidoluysian atrophy, and the spinocerebellar ataxias type 1, 2, and 3, the polyglutamine expansions show both meiotic and mitotic instability. However, previous studies have suggested that the polyglutamine motif within exon 1 of the androgen receptor gene (AR) which expands in cases of spinobulbar muscular atrophy differs in that it is apparently mitotically stable. During linkage analysis in a family with FG syndrome, a rare condition mapped to Xq12 q22.1, we detected the presence of an unusually small AR allele segregating within the pedigree. Sequencing, cytogenetic analysis, and PCR of flanking markers indicate that this allele arose by a somatic contraction of seven CAG repeats in the proband's mother, representing the first report of mitotic instability of an AR CAG repeat of normal size. PMID- 12567415 TI - X-linked recessive chondrodysplasia punctata: spectrum of arylsulfatase E gene mutations and expanded clinical variability. AB - X-linked chondrodysplasia punctata (CDPX1), due to mutations of the arylsulfatase E (ARSE) gene, is a congenital disorder characterized by abnormalities in cartilage and bone development. We performed mutational analysis of the ARSE gene in a series of 16 male patients, and we found mutations in 12 subjects. Clinical variability was observed among the patients, including severe presentations with early lethality in one of them, and symptoms such as cataract and respiratory distress. This indicates that the clinical spectrum of CDPX1, commonly considered a relatively mild form of chondrodysplasia punctata, is wider than previously reported. Different types of mutations were found among the patients examined. Three missense mutations (I80N, T481M, P578S) were expressed in Cos7 cells to study the effects on arylsulfatase E catalytic activity. These mutations caused impaired enzymatic activity suggesting that they are responsible for the disease. Two nonsense mutations, W581X in four patients and R540X in one, were found. One patient showed an insertion (T616ins). In three patients we found deletions of the ARSE gene: in one the deletion involved only the 3' end of the gene, while in two the ARSE gene was completely deleted. PMID- 12567416 TI - Oligoyric microcephaly in a child with Williams syndrome. AB - We report a 19-month-old boy with microcephaly, growth and developmental delay, facial dysmorphisms, and simplified gyral pattern. Magnetic resonance imaging (MRI) examination demonstrated microcephaly with simplified gyral pattern or oligogyric microcephaly. The facial phenotype was interpreted as suggestive of Williams syndrome (WS). Fluorescence in situ hybridization (FISH) analysis performed with an elastin probe revealed a deletion in the chromosomal band 7q 11.23, confirming the clinical diagnosis. To our knowledge, this represents the first patient with WS and oligogyric microcephaly. PMID- 12567417 TI - Sperm segregation patterns by fluorescence in situ hybridization studies of a 46,XY,t(2;6) heterozygote giving rise to a rare triploid product of conception with a 69,XXY,t(2;6)(p12;q24)der(6)t(2;6)(p12;q24)pat karyotype. AB - A blighted ovum diagnosed initially by ultrasound was determined to be a partial hydatidiform mole with a 69,XXY,t(2;6)(p12;q24)der(6)t(2;6)(p12;q24)pat karyotype by cytogenetic analysis. The triploid state arose through dispermy in which both spermatozoa carried rearranged chromosomes, one carrying a balanced translocation through alternate segregation and the other an unbalanced derivative chromosome 6 through adjacent 1 segregation. Segregation analysis of 7,000 spermatozoa from the father was performed with a three-color fluorescence in situ hybridization (FISH) protocol using alpha-satellite 6, telomeric 2p, and telomeric 6q probes. Segregation frequencies of normal and balanced products (alternate segregation), adjacent 1, adjacent 2, and 3:1 were 49.9%, 42.4%, 2.5%, and 4.2%, respectively. The high percentage of alternate segregation is consistent with the knowledge of their preferential outcome. However, the high incidence of adjacent 1 sperm highlights the abnormality risk. Alternate and adjacent 1 segregations (92.3%) accounted for the observed rearranged chromosomes in the triploid. The most viable imbalanced combination would be the one carrying the der(6) chromosome, but since the unbalanced segment comprises 3.6% of the haploid autosomal length (HAL), no risk of a viable imbalanced offspring is indicated. However, an increased likelihood of recurrent miscarriages is likely, and this is confirmed by the couple's two earlier miscarriages. Sperm segregation patterns of translocation carriers determined by FISH can help in ascertaining expected and unexpected karyotypes. The high frequency of adjacent 1 products shows that the presence of the additional derivative chromosome in the partial mole, though rare in occurrence, should be less surprising. PMID- 12567418 TI - CHARGE association and secondary hypoadrenalism. AB - Hypogonadotrophic hypogonadism is a well-described part of the CHARGE association. It has been suggested that more extensive defects of the pituitary function may occur, but this has not been reported in the medical literature. We report a patient with CHARGE association who, in addition to the cardinal manifestations, was found to have hypoadrenalism of pituitary/hypothalamic origin. The patient also has findings typical of the overlap of CHARGE and the 22q11 deletion syndrome, as well as an unusual cervical vertebral abnormality. PMID- 12567419 TI - Teebi hypertelorism syndrome: additional cases. AB - We report on two unrelated Brazilian boys who have craniofacial and digital anomalies resembling those reported with Teebi hypertelorism syndrome. Additional features such as cleft lip and palate, large uvula, atypical chin and abnormal scapulae were observed. PMID- 12567420 TI - Mutation analysis of the MECP2 gene in patients with Rett syndrome. PMID- 12567421 TI - A supernumerary chromosome 20, identified by FISH, in a male with azoospermia cause or coincidence? PMID- 12567422 TI - An unusual family with brachydactyly. PMID- 12567423 TI - New syndrome with generalized lipodystrophy and a distinctive facial appearance: confirmation of Keppen-Lubinski syndrome? PMID- 12567424 TI - Exclusion of the SALL1 gene as a cause of branchio-oculo-facial syndrome. PMID- 12567425 TI - High risk of malignancy in mosaic variegated aneuploidy syndrome. PMID- 12567426 TI - New case of "apple-peel" intestinal atresia and ocular anomalies with mosaic variegated aneuploidy. PMID- 12567427 TI - [Recognition of transient evoked otoacoustic emissions]. AB - OBJECTIVE: To measure the cochlear function more accurately, seeking for a novel method to recognize transient evoked otoacoustic emission (TEOAE) from the recorded signal. METHODS: Time-frequency domain method was used to effectively recognize TEOAE and stimulating artifact by their different short time Fourier transformation (STFT). RESULTS: The STFT of stimulating artifact features decayed rapidly in the air by exponential rule. The frequency distribution of TEOAE was similar to that of the stimulating signal, but the latency and shifting of TEOAE showed a complicated patern. The STFTs of stimulating artifact and TEOAE varied significantly. CONCLUSION: STFT recognition method can separate TEOAE and stimulating artifact effectively so as to improve the accuracy of TEOAE recognition. PMID- 12567428 TI - [Effect of 17 beta-estradiol on activity and gene expression of fibrinolytic factor in human umbilical vein endothelial cell]. AB - OBJECTIVE: To study the effect of 17 beta-estrodial on fibrinolytic factor in human umbilical vein endothelical cell (HUVEC). METHODS: When different concentration of 17 beta-estrodial including 10(-10) mol/L (the group E1), 10(-8) mol/L (the group E2), 10(-6) mol/L (the group E3) were treated in original cultured HUVEC, the changes of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) were measured by spectrophotometric assay, and the level of message RNA were examined by in situ hybridization. The changes of intracellular free calcium concentration([Ca2+]i) in single cells were observed by kinetic observation with Laster Confocal Microscope. RESULTS: The t PA activity both in E1 and E2 were higher than control (P < 0.05 respectively). The t-PA expression in E1, E2, and E3 were increased. The concentration of intracellular free calcium had an elevated and declined curve under the supraphysiological doses of estrodial and declined in incubation with 17 beta eatrodial at 10(-6) mol/L. CONCLUSIONS: These suggeste that the protection of 17 beta-estrodial on cardivascular system is associated with high fibrinolytic activity and t-PA expression as well as with the change of [Ca2+]i. PMID- 12567429 TI - [Effects of growth factors and estrogen on the proliferation and prolactin gene expression in anterior pituitary cells of rats]. AB - OBJECTIVE: Detect the effects of exogenous 17 beta-estradiol (E2), epidermal growth factor (EGF), and transforming growth factor beta 1 (TGF beta 1) on the proliferation and prolactin (PRL) gene expression in primary serum-free cultured anterior pituitary cells in vitro. METHODS: Laser scanning confocal microscopy (LSCM) and in situ hybridization in primary serum-free cultures of rat anterior pituitary cells were employed. RESULTS: After 36 hours incubation of the monolayer with E2(10(-8) mol/L) and EGF(10(-8) mol/L), DNA and PRL mRNA contents in the cells were increased significantly (P < 0.001); and when cells were co incubated with E2 and EGF at the same time, the levels of DNA and PRL mRNA were higher than those treated with E2 or EGF alone (P < 0.01), respectively. TGF beta 1(2 ng/ml) treatment decreased the DNA and PRL mRNA contents significantly (P < 0.001). Its inhibitory effect was reduced at the presence of E2, the DNA and PRL mRNA levels in the cells were higher than those treated with TGF beta 1 alone (P < 0.001), but still lower than E2 alone treatment (P < 0.001). CONCLUSIONS: The results indicate that EGF and TGF beta 1 exerte stimulatory inhibitory effects, on cell proliferation and PRL gene expression in anterior pituitary cells of rats in both basal and E2-induced conditions. EGF and TGF beta 1 may be involoved in the regulation of proliferation and PRL gene expression in anterior pituitary cells in vivo; and also may be correlated with prolactin-secreting tumors formation induced by E2. PMID- 12567431 TI - [Bone mineral analysis of whole body in 292 normal subjects assessed by dual X ray absorptiometry]. AB - OBJECTIVE: To understand whole body bone mineral changes in normal subjects. METHODS: Bone mineral was measured by dual X-ray absorptiometry (DXA) in 292 normal subjects aged 10-79, including 152 females and 140 males. They were divided into age-groups by decade. RESULTS: In males, peak value ages of total bone mineral content(TBMC), total bone mineral density(TBMD), anteroposterior spine BMD(L2-4 BMD) were in the 30-39, 20-39, 20-29 age-groups respectively. In females, they were all in the 30-39 age-group. Peak values of TBMC, TBMD, L2-4BMD were higher in males than those in females. Loss of bone mineral for females was more pronounced than that for males. There was a tendency of increased BMD among the 70-79 age-group for males. Loss of L2-4 BMD was more pronounced than TBMD. CONCLUSIONS: This study provides the normal bone mineral data of whole body for males and females, and the characteristic of changes. PMID- 12567430 TI - [Effects of cyclosporin A combined with T4 on collagen IV production, coll alpha 1 (IV) mRNA and TGF beta 1 mRNA expression in cultured human mesangial cells]. AB - OBJECTIVE: To investigate the effects of single cyclosporin A (CsA), tripcholorlide (T4) or both on renal fibrosis. METHODS: Collagen IV concentrations in the supernatants of human mesangial cells (HMCs) in culture were measured by ELISA. Collagen type IV alpha 1 mRNA (Coll alpha 1 IV mRNA) and transforming growth factor beta 1 (TGF beta 1) mRNA expression were tested by RT PCR. RESULTS: CsA regulated HMC collagen IV production biphysically, i.e., 1.0 microgram/ml CsA increased collagen synthesis while 0.01-0.1 microgram/ml CsA decreased its formation. T4 enhanced collagen IV production in a dose dependent manner. Combination of the two drugs in lower doses could reach the same degree of immunosuppression as a higher dose of CsA did but with less fibrogenesis. Both CsA and T4 up-regulated HMC Coll alpha 1 (IV) mRNA and TGF beta 1 mRNA expression. The degree of Coll alpha 1 (IV) mRNA expression in different groups were in accordance with that of TGF beta 1 mRNA. CONCLUSIONS: Both CsA and T4 regulate extracellular matrix formation which may be mediated by TGF beta 1. Combination of the two drugs in lower dose results in less fibrosis than a higher dose of CsA. PMID- 12567432 TI - [Regulating effects of VEGF and ANG II on the frog's pericardial mesothelium, stomata, and angiogenesis]. AB - OBJECTIVE: To observe the regulating effects of vascular endothelial growth factor (VEGF) and angiotensin II (ANG II) on the frog's pericardium, lymphatic stomata, and angionesis so as to understand their influences on the mesothelial permeability, lymphatic stoma regulation, and myocardial hypertrophy. METHODS: VEGF and ANG II were injected into the frog's peritoneal cavity. The results were obtained by using LM, SEM, and computerized imaging analysis. RESULTS: Scattered distributed pericardial stomata were found on the parietal pericardium of the frog with a few sinusoid mesothelial cells. Since there were no blood vessels in the frog's myocardium, its blood supply was directly from the cardiac chambers flowing into the trabecular spaces of the myocardium. The average diameters of the pericardial stomata in VEGF and ANG II groups were 1.50 and 1.79 microns respectively, which were much larger than those in the control group (0.72 micron, P < 0.01). The average distribution density of the stomata was 8.25/0.1 mm2 and 12.80/0.1 mm2 in VEGF and ANG II groups, which was also much higher than that in the control group (3.57/0.1 mm2, P < 0.01). The sinusoid areas in VEGF and ANG II groups were 2443.95 microns 2/0.1 mm2 and 2121.79 microns 2/0.1 mm2, which was larger than that in the control group (995.08 microns 2/0.1 mm2, P < 0.01). No angionesis were found in the frogs of the experimental groups. CONCLUSIONS: VEGF and ANG II strongly regulate the pericardial stomata by increasing their numbers and openings with larger diameters and higher distribution density. They can also increase the sinusoid areas with the result of the higher permeability of the pericardium, which clealy indicated that VEGF and ANG II can speed up the material transfer of the pericardial cavity and play an important role in preventing myocardial interstitial edema. Yet there is no strong evidence showing angionesis in the myocardium. PMID- 12567433 TI - [Apoptosis inductive effect of homoharringtonine and isoharringtonine on human acute promyelocytic leukemia HL-60 cells]. AB - OBJECTIVE: To study the effects of homoharringtonine and isoharringtonine on the induction of apoptosis on human promyelocytic leukemia HL-60 cells. METHODS: By using flow cytometry, DNA gel electrophoresis, microscopy and electron microscopy. RESULTS: It was demonstrated that homoharringtonine and isoharringtonine can induce apoptosis of HL-60 cells significantly and rapidly. The activity of homoharringtonine was higher than that of isoharringtonine, and it was time and concentration dependent. CONCLUSIONS: Both homoharringtonine and isoharringtonine could induce apoptosis of HL-60 cells in terms of morphology, and DNA ladder assay. The activity of homoharringtonine was much higher than that of isoharringtonine. PMID- 12567434 TI - [Remote effects of local injection of botulinum toxin type A]. AB - OBJECTIVE: To assess the severity and temporal profile of remote effects of botulinum toxin type A, BTX-A (Botox from Allergan Inc, USA and CBTX-A made by Lanzhou Biological Products Institute, China) injected locally on neuromuscular junction. METHODS: Patients had enrolled in a prospective study for their movement disorders, 18 cases with Botox and 22 cases with CBTX-A were studied. Single fiber electromyography (SFEMG) in the extensor digitorum communis muscle or tibialis anterior muscle was performed before and 2-3 weeks, 5-8 weeks, 4-5 months after injection of Botox or CBTX-A. RESULTS: It was measured totally 119 times. Significant increase of jitter was demonstrated 2-3 weeks after injections in both groups and MCD was in direct proportion to dose of injections. Fiber density value increased at the same time or later and lasted until 4-5 months after injections. CONCLUSIONS: Subclinical effects on neuromuscular transmission of remote uninjected muscles after injections of Botox and CBTX-A are demonstrated, which indicates that the toxin spread distantly from the site of injection. SFEMG is valuable for studying the pathogenesis of the remote effects of BTX-A and for avoiding reinjection too frequently. PMID- 12567435 TI - [Construction of eukaryotic vector for human interleukin-15 cDNA and its expression in lung carcinoma cell lines]. AB - OBJECTIVE: Human interleukin(hIL)-15 expression of lung cancer cell lines transfected by rhIL-15 cDNA in vitro was observed. METHODS: The recombinant plasmid pL-IL-15-SN was established by inserting IL-15 cDNA into vector pLXSN at sites of EcoR I and BamH I. The pL-IL-15-SN was transfected into human lung squmosuse carcinoma (PG) cell line and murine lung adenocarcinoma (LA795) cell line, respectively. Positive clones were obtained by the selection in G418 conditioned culture. Bioactivity on rIL-15 was detected by dependant proliferation of CTLL-2 cells in vitro. RESULTS: Three positive PG and four positive LA795 cell clones were obtained respectively. The results of the assays for hIL-15 bioactivity showed that the expression levels of rhIL-15 ranged from 142 to 201 or from 138 to 178U/(ml.10(6)) for positive PG cells or LA795 cells, respectively. CONCLUSIONS: Human and murine lung carcinoma cells transfected with IL-15 cDNA can express hIL-15 with bioactivity. PMID- 12567436 TI - [Cloning and expressing of human papillomavirus 16 E7C subgene]. AB - OBJECTIVE: To develop vaccine against HPV16 is the hotspot in this field, this study was to pave the rudiment for this purpose. METHODS: E7C subgene (encoding aa39 to aa98) was amplified by PCR, and then cloned into pLNCX plasmid. RESULTS: The recombinant pLNCE7C was obtained and identified by gel analysis of the restricted-endonuclease-digested fragments. It was shown that pLNCE7C was expressed in transfected B16 cells. The expression product was identified in the positive cells by Southern blotting and immunohistochemistry method. It was localized mainly in cytoplasm of B16 cells. CONCLUSIONS: The plasmid pLNCE7C containing subgene of HPV16E7C not only remains the antigenesis of E7 gene but also eliminates the transforming activity of it. Meanwhile the expression of pLNCE7C is quite good. Therefore the plasmid pLNCE7C is suitable to develop DNA vaccine against HPV16. PMID- 12567437 TI - [Preliminary examination of a LB-H2O2 substance that inhibit Neisseria gonorrhea growth]. AB - OBJECTIVE: To study the possible mechanism of H2O2-producing lactobacilli inhibited neisseria gonorrhoeae growth. METHODS: H2O2-producing lactobacilli were coincubated with neisseria gonorrhoeae in vitro, LB+ cell were disrupted by sonication, the protein in the lactobacilli lysate was separated by filtration through different MW sizes membranes and was examined for the ability to inhibit N. gonorrhoeae catalase activity. RESULTS: When coincubation medium was at pH 5.0, there was a significant decrease in the gonococcal catalase activity, the lysates of LB+ also effectively inhibited gonococcal catalase activity. This inhibition was retained upon heating of lysate to 100 degrees C for 15 minutes but was lost with proteinase K treatment. CONCLUSIONS: The LB+ may inhibit growth of gonococci by production of catalase inhibitors. PMID- 12567438 TI - [Application of IL-1 beta and 8 mRNA and some other indices in monitoring the activity of ulcerative colitis]. AB - OBJECTIVE: To evaluate the monitoring action of the expression of IL-1 beta, IL-8 mRNA, myeloperoxidase (MPO) and superoxide dismutase (SOD) activity in the active ulcerative celitis (UC). METHODS: Twenty active UC patients, 23 inactive UC patients; and 14 non-UC inflammation patients were selected. Twelve patients who complained of flatulence, abdominal pain and constipation receiving endoscopic examination were used as control. MPO and SOD activity, IL-1 beta and IL-8 mRNA expression (hybridization in situ) were determined on the endoscopic biopsy specimens of all patients. In 20 active UC patients, 14 patients received endoscopic examination and 4 indices detection were done once again after 2 months' treatment with prednisone and SASP. RESULTS: Mucosal MPO activities of active UC patients, inactive UC patients, and non-UC inflammation patients are higher than those of control patients, (19.37 +/- 0.54, 11.59 +/- 1.41, and 12.97 +/- 0.49) U/g tissue vs (9.49 +/- 0.51) U/g tissue (P < 0.01). SOD activities are lower than that of control patients, (5.03 +/- 07,7. 7.66 +/- 0.79, and 6.98 +/- 0.61) U/mg protein vs (8.82 +/- 0.58) U/mg protein (P < 0.05). Mucosal MPO activity of active UC patients is also higher than that of inactive and non-UC inflammation patients (P < 0.01); while SOD activity is lower than them (P < 0.01). After 2 months' medical treatment, MPO activity of 14 active UC patients decreased, (12.61 +/- 0.74) U/g tissue vs (19.31 +/- 0.44) U/g tissue (P < 0.01), while SOD activity elevated (7.44 +/- 0.55) U/mg protein vs (5.10 +/- 1.05) U/mg protein (P < 0.05), compared with that of before treatment. Positive expression of IL-1 beta mRNA appeared in the epithelial and inflammatory cells of all active UC patients, 9 inactive UC patients, and 7 non-UC inflammation patients. While expression of IL-8 mRNA only appeared positively in all active UC patients. In 14 active UC patients, there were no detection of interleukin mRNA expression after 2 months' treatment. CONCLUSIONS: Mucosal MPO, SOD, IL-1 beta, and IL-8 mRNA could be used as 4 indices monitoring the activity of UC. And IL-1 beta mRNA is also helpful to supervise early or late UC activity. PMID- 12567439 TI - [Distribution of 5-fluorouracil in plasma and pancreatic tissue of rats during the regional arterial infusion chemotherapy]. AB - OBJECTIVE: Compare the pharmacokinetic parameters of 5-FU in the regional arterial infusion chemotherapy with those in the intravenous chemotherapy. METHODS: Wistar rats were randomly divided into two groups: the intravenous group and the regional arterial infusion group. HPLC was used to measure the concentration of 5-FU in the plasma and the pancreas tissue. Penetration index was calculated. RESULTS: In the intravenous group, maximum concentration (Cmax) of 5-FU in the pancreas was 8.42 micrograms/g and the clearance time of the pancreas was 50 min. In the regional arterial infusion group, Cmax of 5-FU in the pancreas was 20.00 micrograms/g and the clearance time of the pancreas was 90 min. They were both greater than those of the intravenous group. Rd was 2.42. CONCLUSIONS: The regional arterial infusion chemotherapy is a better way for treating pancreatic carcinoma than the intravenous chemotherapy. PMID- 12567440 TI - [Toxin-binding activity of cyanogen bromide cleaved peptide fragments of the Chinese-cobra serum antitoxic protein]. AB - OBJECTIVE: For further study of the structure-function relationship of the Chinese cobra serum antitoxic protein. METHODS: Cyanogen bromide was used to cleave the peptide bonds which formed by the carboxyl group of Met residues. The cleaved products were ultrafiltrated to remove the small fragments below 10,000 and passed an affinity column packed with Sepharose-4B linked with cobrotoxin as ligand. The molecular weight and N-terminal amino acid residues of the cleaved peptide fragments bounded to the affinity column were then assayed. RESULTS: The cleaved peptide fragments reserved toxin-binding activity were proved to be Lys2 Met485, Lys2-Met275, Lys276-Met485, Pro444-Met603. CONCLUSIONS: This result revealed that all the three domains of CSAP, along the peptide chain, possess the toxin binding activity. PMID- 12567441 TI - [Role of glutathione S-transferase pi in tumor drug resistance]. AB - OBJECTIVE: To establish the cytologic expressing system for the glutathione S transferase pi (GST-pi) cDNA. METHODS: The pSV-GT, a recombinant plasmid containing rat GST-pi cDNA and the pSV-neo, a vector plasmid, were transfected the HeLa cells respectively and then G418 resistant colonies were isolated. The degree of the expression of GST-pi mRNA in the resultant cell lines was estimated by in situ hybridization using Digoxin-labelled probe cDNA. Cytotoxicity of variant anticancer drugs on the transfected cells were estimated by MTT assays. RESULTS: Two subclones, HeLa/pSV-GT, and HeLa/pSV-neo, which transfected respectively by pSV-GT and pSV-neo, were established. HeLa/pSV-GT expressed significantly high degree of the GST-pi mRNA, where as both the HeLa/pSV-neo and the HeLa cell lines showed the similar but negligible degree of the GST-pi mRNA expression. Moreover, HeLa/pSV-GT cells were insensitive to adriamycin, mitomycin C, and cisplatinum as compared to HeLa/pSV-neo. Concentrations of adriamycin producing 50% inhibition (IC50) to HeLa/pSV-GT was 70.13 micrograms/ml, highly elevated 6.78-fold of that of HeLa/pSV-neo, to which IC50 was 10.34 micrograms/ml.IC50 of mitomycin and cisplatinum to HeLa/pSV-GT were 10.95 and 16.52 micrograms/ml respectively, slightly but significantly higher than those to HeLa/pSV-neo, which were 7.48 and 13.70 micrograms/ml. But the cytotoxicity of vincristine on the two cell lines were not significantly different. CONCLUSIONS: These findings suggeste that HeLa/pSV-GT cell line, after being transfected with GST-pi cDNA, acquired drug-resistance property and the overexpression of GST-pi mRNA may contribute to the multidrug resistance of HeLa/pSV-GT cells. This cell line, as a stable cytogenetic system, will contribute more for the observation of GST-pi and drug resistance. PMID- 12567442 TI - [Excretion ratio of lactulose and mannitol as intestinal permeability index in healthy Chinese volunteers]. AB - OBJECTIVE: To determine lactulose (L)/mannitol (M) excretion ratio with high pressure liquid chromatograph with pulsed electrochemical detection (HPLC-PED) in healthy Chinese volunteers. METHODS: Sixty healthy Chinese volunteers enrolled. For each volunteer the L/M permeability study was performed twice by two different doses, as small dose (1 g mannitol and 2 g lactulose) and regular dose (5 g mannitol and 10 g lactulose). The urine were collected during 6 hours after ingestion of the test solution. The M and L concentrations were analyzed by HPLC with pulsed electrochemical detection. RESULTS: The L/M ratio was 0.026 +/- 0.006 in the small doses tests, and 0.036 +/- 0.008 in the large doses tests. CONCLUSIONS: With HPLC-PED detection, a low doses of lactulose and mannitol can be used to study intestinal permeability. Determination of the L/M ratio may be a way of evaluating intestinal barrier function in clinical practice. PMID- 12567443 TI - [Effects of combined treatment with flouride and 1,25-dihydroxyvitamin D3 on the histomorphometry and biomechanical properties of bone in ovariectomized rats]. AB - OBJECTIVE: To evaluate the potential use of a combination of bone formation promoting agents and a differentiation agent of osteoblast as a treatment for postmenopausal osteoporosis. METHODS: The effects of combined and separate administration of flouride-Ca (0.45 ng F- + 13.56 mg Ca2+)/(kg.d) and 1,25 dihydroxyvitamin D (135 pmol/d s.c.) on the histomorphometric parameters in the secondary spongiosa of proximal tibia and biomechanical properties of femoral midshaft in ovariectomized (OVX) rats were examined. RESULTS: Compared with OVX group, OVX + fluoride-Ca group (O + F) and OVX + Fluoride-Ca + 1,25 dihydroxyvitamin D(O + F + D) two kinds of treatment caused 42%, 91% more in %Tb Ar, 14%, 38% more in Tb Wi, 24%, 46% more in Tb N, 26%, 43% less in Tb Sp, 45%, 42% less in % E Pm, 30%, 60% more in Ob Pm respectively. W Wi in O + F treatment had no significant difference from OVX group but 15.2% in O + F + D treatment compared with sham group, MLT increased 16% in O + F group, had no difference from O + F + D. No significant alteration of mechanical strength at femoral midshaft in both O + F and O + F + D was found. There were significant difference in above changes of parameters except % Ob Pm. CONCLUSIONS: Above two treatments could prevent the loss of cancellous bone, reduced the resorption caused by OVX, protected trabecular microarchitecture, and stimulated bone formation significantly. The combined treatment avoided potential mineralization defect caused by F-Ca treatment alone, and two treatments maintained mechanical strength at femoral midshaft. PMID- 12567444 TI - [Phase II clinical study of a new anticancer drug boanmycin]. AB - OBJECTIVE: To evaluate the efficacy of boanmycin (BAM) in patients with advanced cancers. METHODS: A multicenter phase II clinical study on BAM was conducted on 105 cases received BAM as single agent, and 220 cases were treated with combination chemotherapy containing BAM. RESULTS: The total response rate was 35.0% for single agent and 64.2% for combination chemotherapy group in advanced cancers. The response rate for single agent in malignant lymphoma, cancer of head and neck and breast cancer was 66.7%, 65.0% and 37.5%, respectively; and for combination chemotherapy group in malignant lymphoma, breast cancer, head and neck cancer, esophageal cancer and lung cancer was 88.5%, 62.5%, 50.0%, 47.2% and 45.8%, respectively. The main side effects with mild or moderate fever, chill, myalgia, gastrointestinal reactions, skin pigmentation as well as hardening of skin at injection site. CONCLUSIONS: BAM therapy was effective in malignant lymphoma, cancer of head and neck and other cancers. PMID- 12567446 TI - [The inhibitory effect of selenite and other antioxidants on the complement mediated experimental pulmonary alveolitis]. AB - OBJECTIVE: To test the regulatory effect of selenite and other antioxidants on complement PMN activation feedback mechanism mediated inflammatory response in experimantal pulmonary vasculitis. METHODS: Mouse model of pulmonary Arthus reaction vasculitis developed with BSA sensitization and antigen inhalation was used for tests of inhibitory effect of oral administration of sodium selenite, glycyrrhizin flavonoids and/or vitamin E on the incidence of developing vasculitis, which could reflect the modulation of inflammatory response and tissue injury. RESULTS: A remarkable inhibitory effect of selenite or combined flavonoid and vitamin E on the induction of pulmonary vasculitis were observed as the incidence of vasculitis could drop from 100% of the untreated group down to 25% of the antioxidants treated groups, revealing modulation of inflammatory response. CONCLUSIONS: It indicated a newer insight into mechanism of the inflammatory response could give a novel approach to the modulation of inflammatory response. An adjuvant therapy of selenium and antioxidants to the interventions of inflammatory disorders has been expected. PMID- 12567445 TI - [Functional analysis of human A gamma-globin gene promoter with-173 T-->C mutation]. AB - OBJECTIVE: To investigate the effect of the human A gamma-globin gene-173 T-->C mutation on the binding of transacting factors to the promoter and the activity of the promoter. METHODS: Electrophoretic mobility shift assay and transient transfection assay were used in this study. RESULTS: The A gamma-globin gene-173 T-->C mutation decreased the affinity of GATA-1 to the mutant promoter fragment ( 201(-)-158) 96% (P < 0.01) and the binding of Oct-1 to the same DNA fragment 55% (P < 0.05). The activity of the mutant promoter was 2-fold (P < 0.05) as strong as that of the normal one in MELGM979 cells. The mutant and the normal promoters exhibited basically same activities in K562 and Hela cells. CONCLUSIONS: The -173 T-->C mutation decreased dramatically the binding of GATA-1, implicating that GA TA-1 may act as a negative regulator of A gamma-globin gene in adults. The -173 T ->C mutation may enhance the activity of A gamma-globin gene promoter in the adult erythroid cell environment. PMID- 12567447 TI - [Effects of long-term morphine exposure on the cAMP system and c-Fos phosphorylation in differentiated SH-SY5Y cells]. AB - OBJECTIVE: To further understand the effects of long-term morphine exposure on the cAMP system and c-Fos phosphorylation in differentiated SH-SY5Y human neuroblastoma cells. METHODS: Cellular changes of cAMP, PKA and c-Fos were detected by protein competitive conjunction, enzyme activity and isotope incorporation methods respectively. RESULTS: (1) Long-term exposure (2 min-36 h) to morphine (100 mumol/L) could induce the biphasic changes in cAMP contents. Treament for 2 min to 1 h, morphine caused rapid and siginificant decrease of the cAMP level and then gradully recovered and apparently increased at 36 h. At that time, naloxone added to the incubation media caused an overshoot of cellular cAMP; (2) Long-term exposure to morphine could also induce the biphasic changes in cytosolic PKA activity. This is consistent with the changes of cAMP during the chronic treatment of cells with morphine. But no changes were observed in membrane PKA activity; (3) In morphine dependent-like cells decreased c-Fos phosphorylation level was observed. PKA inhibitor could significantly inhibit this change; (4) Concomitant administration of naloxone could block the changes in PKA activity and c-Fos phosphorylation described above. CONCLUSIONS: The up regulation of cAMP system in differentiated SH-SY5Y cells may be involved in the development of morphine dependent and in morphine dependent-like SH-SY5Y cells and PKA was suggested to regulate c-Fos dephosphorylation through activating phosphatase and then activate some genes transcription, which might be one of the important mechanism regardingas cellular adaptive responses underlying dependence to opioid drugs. PMID- 12567448 TI - [Aspirin inhibited the adhesion of platelets to neutrophil in patients with acute myocardiac infarction]. AB - OBJECTIVE: To investigate the effect and mechanism of aspirin(ASA) on platelet neutrophil adhesion. METHODS: Using rosette forming assay to observe the effect of ASA on the binding of platelets to neutrophil and radioimmunoassay to observe the effect of ASA on the thrombin-induced expression of GMP-140 on the surface of human platelets. RESULTS: Patients with acute myocardial infarction showed a significant increase in neutrophil-platelet adhesion (63.3 +/- 7.8)%, (n = 20, P < 0.001) as compared with normal subjects(16.5 +/- 2.6)% (n = 20). Thrombin promoted adhesion by increasing the expression of GMP-140 on the surface of human platelets. When the concentration of thrombin was 0.5 U/ml, the rate of neutrophil-platelet adhesion and the number of molecular of GMP-140 on the surface of platelet was (64.2 +/- 4.2)%, and (1.80 +/- 0.20) x 10(3) per platelet respectively. At high concentration, ASA significantly inhibited thrombin (0.5 U/ml) stimulated platelets binding to neutrophils and expressing GMP-140 on their surface. When the final concentration of ASA was 500,5000 micrograms/ml, the ratio of thrombin-stimulated platelets binding neutrophils was (34.7 +/- 3.8)%, (21.2 +/- 3.6)% respectively (n = 20, P < 0.01); the number of molecular of GMP 140 expressing on the surface of platelet was (1.02 +/- 0.24) x 10(3), (0.68 +/- 0.18) x 10(3) per platelet respectively (n = 9, P < 0.001). CONCLUSIONS: It contributes to the antithrombosis of ASA that inhibiting platelet-neutrophil adhesion by down expression of GMP-140 on the surface of human platelets. PMID- 12567449 TI - [Treatment of late thoracolumbar fracture by transpedicular decompression using "PUMC trephine" 26 cases report]. AB - OBJECTIVE: To introduce a new operation to treat late thoracolumbar fracture cases with neurological deficit or with kyphosis. METHODS: 26 cases were treated with "PUMC trephine" (designed by professor Ye Qibin). RESULTS: The kyphosis was (29.7 +/- 14.5) degrees Cobb's angle in pre-operation and (10 +/- 11.8) degrees in post-operation (P < 0.01). The angle of correction was (19.7 +/- +9.9) degrees. The height of the anterior edge of traumatic-vertebra was (16.3 +/- 5.7) mm in pre-operation and (22.2 +/- 6.3) mm in post-operation. The corrected height was (6.5 +/- 3.2) mm. The neurological deficit also got a better recovery in 17 of 19 cases, 12 of 15 cases obtained complete cure of incontinence. 11 of 15 case had sexual function recovery after following up 1-9 years (average 5.2 years) cases. CONCLUSIONS: Using "PUMC trephine" procedure was a new, simple, safety and less-traumatic technique for treatment of late thoracolumbar fracture through posterior approach. PMID- 12567450 TI - [Protection of PNH red blood cells from lysis by transferring GPI-anchored protein vesicles of normal human RBCs in vitro]. AB - OBJECTIVE: To observe the transfer of GPI-anchored protein CD59 from vesicles of normal RBCs to paroxysmal nocturnal hemoglobinuria(PNH) RBCs can correct the susceptibility to complement attack in vitro. METHODS: Vesicles released from normal RBCs under ATP depletion or storage, are rich in CD59. PNH CD59- RBCs were separated by elution from immunoaffinity column bound with monoclonal antibody CD59, and then incubated with normal RBCs vesicles. The content of CD59 in vesicles and RBCs was detected by Western blot and flowcytometric analysis respectively and hemolysis of PNH CD59- RBCs was detected by cobra venom factor hemolysis test. RESULTS: The fluorescence intensity of PNH CD59- RBCs were increased from (1.68 +/- 0.57)% to (58.42 +/- 8.25)% and the hemolysis was significantly decreased from (27.13 +/- 9.69)% to (19.49 +/- 7.61)% after incubation with vesicles prepared from normal donors (P < 0.05), while normal RBCs had no obvious change before and after the same treatment. (P > 0.05). CONCLUSIONS: It seems that CD59 protein molecules could be transfered from normal RBCs vesicles to PNH CD59- RBCs and retaining the complement regulatory function. CD59- RBCs could be rendered less liable to hemolyze during complement attack. PMID- 12567451 TI - [The effects of sex hormone on blood vessels]. AB - OBJECTIVE: The protective mechanism of sex hormone on blood vessels was investigated to provide evidences for clinical use of hormone replacement therapy in postmenopausal women. METHODS: To observe the effects of 17 beta-estradiol (E2), progesterone(P), testosterone (T) and E2 combined with P on the relaxation of umbilical arterial rings (UAR) as well as production of nitric oxide (NO) in cultured umbilical endothelial cells(HUVEC) in vitro. RESULTS: The results showed that 10(-9)-10(-6) mol/L E2 dilation of UAR, but 10(-5) mol/L E2 showed vague effect. P or T had no relaxation effect, but releasing nitric oxide. When in combined trial P at 10(-5) mol/L inhibited the relaxation effect of E2 at 10(-7) 10(-5) mol/L but didn't inhibit E2 10(-9) mol/L. In HUVEC 10(-9)-10(-5) mol/L E2 stimulated the release of nitric oxide when cells incubated for 10 min and 30 min, but did not influence the NO release at 60 min. P and T have different effects on the release of nitric oxide at different concentrations and incubation time. P also inhibited the NO release induced by E2. CONCLUSIONS: The results indicated that P actually inhibited the relaxation response and induction of NO release by E2, especially at the high concentration. Therefore, we suggested that to use low doses of E2 combined with low dose P for hormone replacement therapy in prevention and treatment of cardiovascular diseases in postmenopausal patient. PMID- 12567452 TI - [Structural disorder and conformation difference in the organic molecular crystals]. AB - OBJECTIVE: Investigating multiple configuration nature of natural organic molecules through study on atom disorder in crystal state molecules. METHODS: Three-dimensional determination for four natural organic compound samples by using single crystal X-ray diffraction. RESULTS: Through disorder analysis on molecules in crystal state, we have found that the disorder of some atoms may cause conformation difference for molecules, bring about multiple conformation in crystal state hence reducing the symmetry of molecules in crystal cells. CONCLUSIONS: There can be two or three molecules, even two different compounds in an asymmetrical unit of structure cell because of partial disorder or configuration difference in organic molecule crystals. PMID- 12567453 TI - [Efficient, stable expression of human interleukin-15 cDNA in Chinese hamster oval cells]. AB - OBJECTIVE: To express the human interleukin 15(IL-15) cDNA in Chinese hamster oval (CHO) cells, which would benefit to the further research in the biological activities and the clinical applications of IL-15. METHODS: The entire human interleukin-15(IL-15) coding region, deleted of all the uncoding fragments, was amplified by PCR. The amplified products were subcloned into the EcoR I and Xba I sites of the pcDNA3 plasmid, forming the recombinant eukaryotic expressing vector, pcDNA3-IL-15, which was then identified by the Bg1 II enzymatic digestion, PCR amplification and the sequence analysis. The construct was transfected into CHO cells by means of lipofectamine, followed by a series of determinations to screen the positive cell colonies, such as RT-PCR, SDS-PAGE, ELISA and CTLL-2 proliferation assay. RESULTS: 8 positive cell colonies highly expressed human IL-15 were obtained after G418 selection. The bioassay showed that the mean activities of the supernatants from the CHO-IL-15 cells were (318.54 +/- 32.76) U/(10(6) cells.d) and maintained stable after 6 months culture. CONCLUSIONS: Human IL-15 cDNA was expressed efficiently and persistently in CHO cells. PMID- 12567454 TI - [Detection of telomerase activity in gastric cancer and adjacent tissues]. AB - OBJECTIVE: To study the telomerase activity in chinese gastric cancer and their adjacent tissues and to explore the possibility of telomerase as a tumor biological marker. METHODS: Telomerase activity was analyzed in 42 primary gastric cancers and their adjacent tissues and 1 benign gastric lesion by a PCR based telomerase repeat amplification protocol assay. RESULTS: The result showed 88.1% (37/42) gastric cancerous specimens displayed telomerase activity. However, only 2 of 42 tissue samples adjacent to the gastric cancer showed telomerase activity. However our results indicated that 1 benign gastric lesion sample expressed telomerase activity. CONCLUSIONS: The results indicated that the detection of telomerase activity might be a useful method in the diagnosis of gastric cancer. PMID- 12567455 TI - [The effect of cAMP on the transcription and regulation of angiotensin II type 1 receptor gene in cardiomyocytes of neonatal rats]. AB - OBJECTIVE: To clarify the effect of cAMP on the transcription of angiotensin II type 1 (AT1) receptor mRNA in cardiomyocytes. METHODS: Cardiomyocytes from SD neonatal rats were cultured in DMEM containing 10% fetal bovine serum (FBS) with 0.1 mmol/L 5-bromodeoxyuridine (Brdu). At day 5-7, primary cultured cells were used. FBS and Brdu were deprived 24 h before the stimulation. The levels of AT1 mRNA in primary cultured cardiomyocytes were examined by RT-PCR. RESULTS: Stimulation of forskolin (30 mumol/L) plus 3-isobutyl-1-methyl-xanthine (IBMX, 30 mumol/L) which induced the accumulation of cellular cAMP decreased the expression of AT1 receptor mRNA to (64.4 +/- 20.3)% of control at 6 h treatment; while it increased the expression of AT1 receptor mRNA to (209.6 +/- 24.0)% of control at 48 h treatment. CONCLUSIONS: cAMP regulated the transcription of AT1 receptor gene. PMID- 12567456 TI - [Influence of radiation on the blood-brain barrier and optimum time of chemotherapy]. AB - OBJECTIVE: A pilot study of the destructive effects of radiation on the blood brain barrier (BBB) and blood-cerebrospinal fluid barrier (BCSFB) were carried out on 29 patients. METHODS: (1) First BBB with localized and limited brain tumors was examined by 99mTc-GH imaging count/pixel, data was obtained from the unirradiated, irradiated, and tumor areas of before and after radiotherapy of 20 40 Gy respectively. (2) The BCSFB was studied quantitatively by observing the effect of methotrexate (MTX) permeation into the CSF before, during, and after brain irradiated after i.v. injection of MTX. RESULTS: (1) the BBB in the unirradiated area outside the radiation portal was not changed, (2) the degree of destructive effect on the BBB in the irradiated normal area and on the BCSFB were directly proportional to radiation doses. CONCLUSIONS: When the permeation of BBB and BCSFB irradiated, was enhanced the degree of destructive effect were enhanced proportionally with irradiation doses. It was that the BBB irradiated can be recovered at least partially after irradiation of 3 weeks proved. After a dose of 20 Gy irradiation, the BCSFB would gradually open. As compared with the pre radiation data, the permeability of MTX increased by 1.05-1.3 times. So, it was advisable to give chemotherapy after 20 Gy irradiation. PMID- 12567457 TI - [Autoantibodies to the heterogeneous nuclear ribonucleoproteins and the autoimmune diseases]. AB - Autoantibodies to the heterogeneous nuclear ribonucleoproteins (hnRNP) have a great role in the diagnosis of rheumatic diseases. hnRNP-A/B proteins are the main autoantigens in Rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and mixed connective tissue disease (MCTD). Anti-RA33 autoantibodies which target the hnRNP proteins A2, B1, B2 (the 'RA33 complex') help to the diagnosis of RA at early stage. Therefore, anti-hnRNP autoantibodies are not only valuable diagnostic markers but may also allow deep insights into the pathogenetic mechanisms of rheumatic diseases. PMID- 12567458 TI - [Sequence analyzing and genotyping of the gene encoding glutamate rich protein of geographically different Plasmodium falciparum isolates obtained from different malaria endemic areas]. AB - OBJECTIVE: To sequence a gene encoding GLURP and identify the genotypes of geographically different Plasmodium falciparum isolates from Yunnan and Hainan Provinces, China. METHODS: The gene of R2 repeat region of GLURP was amplified by the nested polymerase chain reaction and cloned into T-vector. The nucleotide sequence of the GLURP gene was determined using automatic sequencer (dideoxy chain termination method), and analyzed by DNA Star software. RESULTS: At least 7 different GLURP genotypes ranging from 600 bp to 1,500 bp were found in different P. falciparum isolates from Yunnan and Hainan Provinces. R2 region of GLURP gene consisted of several repeat units, each was composed of 19-20 residues which were shown to be highly conserved. The GLURP gene was also size polymorphic due to differences in the number of repeat units, whereas the repeat sequence was conserved. Sequence analysis showed that DNA sequences and deduced amino acid sequences were highly homologous among the geographically dispersed isolates or various isolates from the same geographical region. No obvious differences were found in the GLURP gene sequences among geographically different isolates. CONCLUSION: The GLURP gene of geographically different P. falciparum isolates is highly conserved and size polymorphic, being useful in searching for malaria vaccine candidate antigen and developing a genotyping method for malaria research. PMID- 12567459 TI - [A case with subdural hydrops caused by cerebral cysticercosis]. PMID- 12567460 TI - [Effect of pyronaridine, mefloquine and quinine on artesunate-sensitive and artesunate-resistant Plasmodium falciparum]. AB - OBJECTIVE: To compare the sensitivity of artesunate-sensitive and artesunate resistant P. falciparum to pyronaridine, mefloquine and quinine and to understand the effect of artesunte combined with the above-mentioned 3 drugs respectively on artesunate-resistant P. falciparum. METHODS: Rieckmann's in vitro miorotechnique was used. RESULTS: The ID50 values of pyronaride, mefloquine, quinine and artesunate were 59.0, 69.7, 283.8 and 9.6 nmol/L to artesunate-sensitive P. falciparum; the ID50 of the 4 drugs mentioned above were 170.6, 63.2, 272.4 and 85.1 nmol/L to the artesunate-resistant P. falciparum, respectively. In artesunate pyronaridine combination, the ID50 values were 1/3.7 (22.8/85.1) and 1/4.7 (36.6/170.6) of the 2 drugs singly used. In artesunate mefloquine combinaton, the ID95 is 1/125 (3.2/400) and 1/16 (80/128) of the 2 drugs singly used, respectively. CONCLUSION: The artesunate-resistant P. falciparum isolate has no cross resistance to mefloquine and quinine. When artesunate was used in combination with the 2 drugs mentioned above respectively, the efficacy proved to be enhanced. PMID- 12567461 TI - [Studies on the sensitivity of Plasmodium falciparum to pyronaridine/sulfadoxine/pyrimethamine in vitro]. AB - OBJECTIVE: To develop an in vitro method for the assessment of drug response in P. falciparum to pyronaridine/sulfadoxine/pyrimethamine (PND/S/P). METHODS: The PND/S/P microtest plate was designed, Rieckmann in vitro microtest(WHO standard kit) was used to test the sensitivity of P falciparum in continuous culture (FCC1/HN strain), and the data obtained were analyzed using a computer programme. Fractional inhibitory concentration (FIC) was calculated to test the possible synergy between PND and S/P. RESULTS: The effect of the PND/S/P plates was fairly stable and the ED50 values of pyronaridine, sulfadoxine and pyrimethamine were 0.11, 215.12 and 2.9 pmol, respectively. The FIC obtained confirmed the synergism between PND and S/P. CONCLUSION: The in vitro method can be used to assess the sensitivity of P. falciparum to pyronaridine/sulfadoxine/pyrimethamine. PMID- 12567462 TI - [Relationship between hemolymph phenol oxidase and melanization of oocysts of Plasmodium yoelii in Anopheles stephensi]. AB - OBJECTIVE: To explore the relationship between the hemolymph phenol oxidase and melanization of oocysts. METHODS: Anopheles stephensi-Plasmodium yoelii system was used to determine the activity of monophenol oxidase (MPO) and o-diphenol oxidase (o-DPO) in the hemolymph collected from 4 groups of mosquitoes by polyacrylamide gel electrophoresis (PAGE) followed by density scanning. The 4 groups of mosquitoes were: non-blood-fed (N), normal-blood-fed (B), infected blood-fed (I) and nitroquine-administrated (D), respectively. RESULTS: No significant difference was found in the activities of MPO and o-DPO between groups N and B. The activities of MPO and o-DPO were not obviously modified in group I, but were significantly increased on day 10 and decreased on day 15 after blood feeding in the group D as compared with those in the groups N and B. CONCLUSION: The alteration in the mosquito hemolymph PO activity coincided at each time point with the melanization of Plasmodium yoelii oocysts. PMID- 12567463 TI - [Studies on human cytokine responses before and after praziquantel chemotherapy in an endemic area of schistosomiasis japonica]. AB - OBJECTIVE: To observe the cellular immune responses in a population of an endemic area of schistosomiasis japonica and the influence of praziquantel treatment. METHODS: Blood was taken from 129 residents (64 cases were egg-positive, 65 cases were egg-negitive) of an endemic area of Poyang Lake before and 45 days after praziquantel treatment. Cytokines induced by the schistosome soluble egg antigen (SEA) and soluble worm antigen preparations (SWAP) in the peripheral blood cells including IL-5, IL-10 and IFN-gamma were measured. RESULTS: Among 129 cases, the cytokine levels were found much higher in egg negative individuals than in egg positive individuals. The cytokine levels induced by both antigens were increased significantly after praziqantel treatment especially IL-5 and IFN-gamma. CONCLUSION: The cellular immune responses in the population in schistosomiasis japonica endemic area exhibited a general trend of down-regulation and were elevated significantly after praziquantel treatment. PMID- 12567464 TI - [Survey of human hair follicle mite infection in the students in Jinan Command School]. PMID- 12567465 TI - [Studies on risk factors for liver fibrosis of schistosomiasis japonica]. AB - OBJECTIVE: To explore the risk factors for schistosomal liver fibrosis. METHODS: 192 hepatitis-negative patients with schistosomiasis were selected and divided into 3 groups according to the result of B ultrasound examination, that is, grade 2 and 3 fibrosis group(81 patients), grade 1 fibrosis group (61 patients) and control group (non-fibrosis, 50 patiets). The univariate and multivariate ordinal regression model was made to analyse the possibly harmful factors influencing the liver damages of these patients. RESULTS: Four factors were found to be positively associated with schistosomal liver fibrosis. They were: number of treatments of schistosomiasis (OR = 1.75), interval of schistosomal infection(OR = 1.40), history of drinking wine (OR = 1.95) and familial history of advanced schistosomiasis (OR = 2.11). CONCLUSION: Patients with repeated schistosome infection, long duration of schistosomal infection, long history of drinking and familial history of advanced schistosomiasis had higher risk for liver fibrosis than schistosomiasis patients without these factors. PMID- 12567466 TI - [The diagnostic value of combined detection of circulating antigens and antibodies in urine of patients with schistosomiasis japonica]. AB - OBJECTIVE: To assess the value of detecting circulating antigens and antibodies in urine as a noninvasive method for the diagnosis of schistosomiasis. METHODS: A sandwich ELISA and ELISA using McAb were applied to detect circulating schistosomal antigens and specific antibodies in the urine of patients with acute and chronic schistosomiasis. RESULTS: When the urine samples from 10 cases of acute schistosomiasis and 61 cases of chronic schistosomiasis were examined, the positive rates of circulating antigens and specific antibodies were 60%, 40%, and 80%, 60.1%, respectively; when both detection was combined, the positive rates were 100% and 71.7% respectively, whereas a false positive rate of 3% of CAg or CAb was detected in the urine of 100 normal controls. CONCLUSION: The detection of circulating antigen in urine is a practicable and noninvasive method for the diagnosis of schistosomiasis. PMID- 12567468 TI - [Preliminary study on the mollusciciding effect of N-pyridine 3-N'-phenylurea hydrochloride]. PMID- 12567467 TI - Construction and analysis of cDNA library of Necator americanus third stage larvae. AB - OBJECTIVE: To obtain the genetic information on Necator americanus and to search for the purpose genes. METHODS: mRNA was isolated from the third stage larvae of Necator americanus maintained in hamsters. Double strand cDNA was synthesized and ligated to lambda ZAPII vector to construct the cDNA library. Expressed sequence tages (ESTs) were obtained by single pass sequencing of randomly isolated cDNA clones from the established library. RESULTS: A cDNA library of N. americanus was successfully constructed with high recombinant efficiency. The titer of unamplified library was 1 x 10(7). The insert size was about 750-3,000 bp. Of 11 ESTs obtained from the library, 7 have a significant homology with certain functional genes. CONCLUSION: A high quality and high representative cDNA library of N. americanus was constructed at the first time and some functional genes were identified from the library by ESTs. PMID- 12567469 TI - [Cloning and sequence analysis of a gene encoding amastin from Leishmania major]. AB - OBJECTIVE: To clone a gene encoding surface protein from Leishmania major. METHODS: Using T. cruzi amastin DNA sequence as a reference, computer search was done on GenBank and dbEST databases by using BLAST path. A Leishmania major DNA library has been constructed and screened by in situ colony hybridization. RESULTS: A 309nt DNA fragment from Leishmania major was found in dbEST. Leishmania major DNA library was screened using specific primers synthesized according to 309 nt DNA sequence, and a full-length coding sequence for Leishmania major amastin was cloned. The coding sequence consisted of 552 nt, and translated into 183 amino acid residues. The homology is 23.5% at amino acid sequence level between Leishmania major and T. cruzi amastins. CONCLUSION: A full length amastin coding gene for Leishmania major has been cloned. PMID- 12567470 TI - [Malaria situation in Anqi County, Fujian province in 1950-1997]. PMID- 12567471 TI - [The protective immunity in mice immunized with FhGST or AsGST against Schistosoma japonicum cercariae]. AB - OBJECTIVE: To study the protective immunity against Schistosoma japonicum (Sj) cercariae challenge in mice immunized with FhGST or AsGST. METHODS: There groups of Kunming mice were immunized three times with rSjGST, FhGST and AsGST, respectively. Then 30 cercariae of Schistosoma japonicum were given per mouse by abdominal skin inoculation. Six weeks later, all mice were killed to collect adult worms, liver, spleen and large intestine for worm count and egg count. RESULTS: The worm burden was reduced by 27.8%-36.4% in the three immune groups compared with the two control groups (P < 0.05). rSj26GST, FhGST and AsGST significantly decreased the number of eggs deposited in the tissues. CONCLUSION: FhGST and AsGST could induce protection against S. japonicum cercariae infection. PMID- 12567472 TI - [A case of gongylonemiasis]. PMID- 12567473 TI - [Cloning and expression of Cysticercus cellulosae antigen cC1 in E. coli]. AB - OBJECTIVE: To clone and express Cysticercus Cellulosae antigen cC1 in E. coli. METHODS: cC1 cDNA fragment was cloned to BamHI and PstI sites of pGEM-3Z vector. After alteration of the restriction sites, the fragment was cloned to EcoRI and XhoI sites of pGEX-5T with a synthetic linker to construct recombinant expression vector pGEX-5T-cC1. RESULTS: The clone produced the largest yield of cC1 protein expression when incubated in 2YT culture medium for 3 h or induced by IPTG for 6 h. Detected by scanning optical densitometry, cC1 constituted 57% of the total bacterial proteins. Western blotting analysis revealed that the GST-cC1 fusion protein exhibited a specific reactive band. CONCLUSION: High level expression of Cysticercus cellulosae antigen cC1 was obtained in E. coli. PMID- 12567474 TI - [Immune response in Pneumocystis carinii pneumonia in rats]. AB - OBJECTIVE: To study the inflammatory immune response to Pneumocystis canii pneumonia(PCP) in rats induced by glucocorticoid(GC). METHODS: The model of PCP was set up by injecting GC subcutanously to SD rats. Lymphocyte proportion in peripheral blood, CD4+/CD8+ T cell ratio of PBL and lymphocyte proportion in the BALF were measured. The levels of sIL-2R and TNF-alpha in the BALF were detected. RESULTS: 1. After the rats were immunospressed, the lymphocyte proportion in the peripheral blood and CD4+/CD8+ ratio of PBL, and the lymphocyte proportion in the BALF were decreased, and the levels of sIL-2R, TNF-alpha in BALF were reduced. 2. The lowest levels of TNF-alpha in BALF and CD4+/CD8+ T cells of PBL were observed in PCP group; 3. The lymphocyte proportion in the BALF was significantly higher in PCP group than in PC negative group. CONCLUSION: The reduction in the level of TNF-alpha and CD4+/CD8+ T cell ratio in rats treated with GC might result in PCP infection under immunosuppressive condition. PMID- 12567475 TI - [Detection of circulating antigen in urine from mice infected with Toxoplasma tachyzoites by SPA-ELISA]. AB - OBJECTIVE: To explore a simple and convenient immunoassay for early diagnosis of Taxoplasma infection. METHODS: Urine samples collected from three groups of mice infected with different doses of tachyzoites were detected for Taxoplasma circulating antigen (TCA) by dot-ELISA using HRP-SPA as a second antibody (SPA ELISA). RESULTS: Taxoplasma circulating antigens were detected in all three groups of infected mice in contrast with the normal control group. Taxoplasma circulating antigen was detected on d6 and d3 after infection in mice of light- and moderate-infection groups, respectively. CONCLUSION: SPA-ELISA is a simple and convenient method for early immunodiagnosis of recent Taxoplasma infection. PMID- 12567476 TI - [Qualitative and quantitative comparison of three agglutination tests for detecting Toxoplasma gondii antibodies]. AB - OBJECTIVE: To evaluate the diagnostic value of three agglutination tests used in three countries for detection of antibodies to Taxoplasma gondii. METHODS: A total of 288 human serum samples were assayed using modified agglutination test (MAT-1, using selfmade antigen), latex agglutination test (LAT, using Japanese Kit) and modified agglutination test (MAT-2, using French antigen). RESULTS: The positive rates of MAT-1 (> or = 1:20), LAT (> or = 1:32) and MAT-2 (> or = 1:20) were 9.7% (28/288), 8.9% (10/112) and 8.1% (17/210), respectively. No significant statistical difference was found among these positive rates (chi 2 = 0.392, P > 0.05). High agreements were found between MAT-1 and LAT (93.7%), LAT and MAT-2 (94.5%), and MAT-2 and MAT-1 (97.3%). Significant correlation were demonstrated in MAT-1 and LAT (r = 0.613), LAT and MAT-2 (r = 0.551), and MAT-2 and MAT-1 (r = 0.841), p < 0.001. CONCLUSION: The detection efficiency of the three agglutination tests is in good agreement and could alternatively be used for the diagnosis of toxoplasmosis. PMID- 12567479 TI - [Progress in the research on schistosome genome]. PMID- 12567477 TI - [Practicability of IFAT using Plasmodium cynomolgi and Plasmodium falciparum antigens in different malarious areas]. AB - OBJECTIVE: To compare the practicability of IFAT in different malarious areas using Plasmodium cynomolgi(P.c.) and Plasmodium falciparum(P.f.) antigens. METHODS: This survey was carried out in Yaliang Township of Sanya City, Hainan Province, where a mixed malaria is endemic, and in Tongbo County, Henan Province where only vivax malaria is endemic, and in Weihui City, Henan Province where vivax malaria has been under effective control since 1994-1998. RESULTS: In Yaliang Township, 310 blood samples were examined, the antibody positive rates with P.c. and P.f. were 37.4% and 31.3%, respectively, the rate of coincidence being 83.9%. In Tongbo County, 300 blood samples were examined. The antibody positive rates with P.c. and P.f. were 23.0% and 9.7%, respectively (P < 0.01). Another 245 blood samples from children were examined in Weihui City and the antibody positive rates were below 1% with two antigens, while the positive antibody rate was 3.3% with P.f. antigen. CONCLUSION: Both P.f. and P.c. antigens could be used in malaria antibody surveillance in mixted endemic areas, while in vivax malaria endemic areas, P.c. antigen was recommended. PMID- 12567478 TI - [Efficacy of surgical treatment of hydrocele in 167 cases with filariasis bancrofti]. PMID- 12567480 TI - [Diagnosis, treatment and prevention of Toxoplasma infection during pregnancy]. PMID- 12567481 TI - [Compliance analysis of the residents with mass medical treatment in areas highly endemic for schistosomiasis]. PMID- 12567482 TI - [Survey of Toxoplasma infection in patients with thyroid diseases]. PMID- 12567483 TI - [Preliminary establishment of data-managing information system of filariasis]. PMID- 12567484 TI - [Six-year survey of malaria in the agriculture reclamation system in Hainan]. PMID- 12567485 TI - [Changes in electrocardiogram in cases with allergic urticaria by Encleidae]. PMID- 12567486 TI - [Diagnosis and surgical treatment of brain-stem vascular malformations]. AB - OBJECTIVE: To study the various aspects of the brain-stem vascular malformations, including the histopathology, clinical presentation, neuroimaging, techniques of surgical operation. METHODS: A retrospective analysis of 100 patients with 105 brain-stem vascular malformations had underwent microsurgery from 1987 to 1997 in the institute. RESULTS: Most brain-stem vascular malformations are cavernoma, some of them are not easily differentiated from telangiectasis. Meanwhile, they could also associate with venous malformation. In our group, 67% patients suffered from hemorrhage more than one time. The average number of bleeding in female is higher than in male's. There was no operative mortality. The outcome was improved and stable in 65% patients. Ninety-two patients have followed-up half a year to 11 years. Rebleeding after operation occurred in 4% cases. CONCLUSIONS: Active microsurgery should be adopted carefully for the patients with symptomatic brain-stem vascular malformations, while those with asymptomatic lesions need long-term clinical observations and neuroimaging follow-up. PMID- 12567487 TI - [Overexpression of Bcl-2 inhibited T lymphocyte apoptosis mediated by CD3 epsilon molecule]. AB - OBJECTIVE: To study the role of bcl-2 gene overexpression in T lymphocyte apoptosis mediated by CD8 epsilon chimera molecule. METHODS: The cells expressed CD8 epsilon chimera molecule were transfected with a bcl-2 expression vector. The cells with Bcl-2 overexperssion were selected by G418 and limited dilution, and then treated with anti-CD8 antibody. The cell death was evaluated with flow cytometry. RESULTS: It was shown that there were only 2.6% of cells with Bcl-2 overexpression died by apoptosis. Comparing with Jurkat and TJK cells without Bcl 2 expression, in which there were 6.1% and 33.7% of cell death respectively, suggesting that Bcl-2 overexpression remarkbly inhibited apoptosis of the T lymphocytes mediated by CD8 epsilon chimera molecule. CONCLUSIONS: Bcl-2 overexpression could block T lymphocyte apoptosis mediated by CD3 epsilon molecule and the mechanisms remained to be clarified in detail. PMID- 12567488 TI - [Construction and in vivo expression of the recombinant adenovirus containing human pro-UK cDNA]. AB - OBJECTIVE: To construct the recombinant adenovirus which contained human pro-UK cDNA and to provide experimental basis for clinical gene therapy. METHODS: The recombinant plasmid pCA14-pro-UK and the plasmid pJM17 were cotransfected into cultured 293 cells and the replication-deficient adenovirus were harvested, identified, propagated and titered. The recombinant virus were then transferred into balloon-injured femoral arteries of rabbits and the pro-UK expression was detected by immunohistochemistry and Western Blot. RESULTS: PCR amplification and in vitro transfection of A549 cells confirmed that pro-UK had been inserted into adenovirus genome correctly, and immunohistochemistry and Western blot analysis showed that pro-UK expressed evidently in the balloon-injured sites of the rabbit femoral arteries. CONCLUSIONS: The recombinant adenovirus that constructed could express pro-UK cDNA in vivo successfully. PMID- 12567489 TI - [Cloning of full-length Dnmt3b cDNA and its alternative splicing isoforms in mouse embryo]. AB - OBJECTIVE: To clone the novel full-length cDNA related to mouse embryonic development. METHODS: Starting from a new expression sequence (clone 2 #) out of a mouse embryonic cDNA library, its full-length cDNA were obtained by rapid amplification of cDNA ends with polyA RNA isolated from mouse 8-9 day embryonic tissue as template, combined with screening of mouse embryonic cDNA library by PCR. Based on the sequences at the two ends of the cDNA, the primers for long range RT-PCR were designed to confirm its factuality. RESULTS: The cDNA isolated in our hands was one of the alternative splicing isoforms of mouse de novo DNA cytosine 5'-specific methyltransferase gene (Dnmt3b) reported in July, 1998. A new alternative splicing region upstream of its initiate codon was found 5' forwards 152 bp over the Dnmt3b cDNA sequence. Eight alternative splicing isoforms of Dnmt3b from mouse embryonic tissue were cloned (Genbank accession number are AF151969, AF151970, AF151971, AF151972, AF151973, AF151974, AF151975, AF151976 respectively). CONCLUSIONS: The 4 kb full-length Dnmt3b cDNA was isolated. Dnmt3b may have novel exons at its 5' upstream and present a complex alternative splicing pattern during mouse embryonic development. PMID- 12567490 TI - [6A8 cDNA encodes A type of alpha-mannosidase]. AB - OBJECTIVE: To find the nature of the protein encoded by 6A8 cDNA. METHODS: In the light of that the ligand of Con A is mannoses and that the function of alpha mannosidase is trimming of mannoses in the oligosaccharide chains of glycoproteins, COS-7 cells and CNE-2L2 cells, a cloned cell line of human nasopharyngeal carcinoma, were transfected with pRc/CMV-sense 6A8 cDNA or pRc/CMV antisense 6A8 cDNA and the Con A binding to the cells was examined upon a lazer scan confocal microscope. RESULTS: The data demonstrated that with transfection of pRc/CMV-sense 6A8 cDNA the binding of Con A to the cells was decreased, but the staining of the cells with mAb 6A8 was enhanced, and vise versa for the cells with transfection of pRc/CMV-antisense 6A8 cDNA. CONCLUSIONS: The protein encoded by 6A8 cDNA might be a type of alpha-mannosidase recognized by mAb 6A8. PMID- 12567491 TI - [Study on GMA-DNA adducts in vivo]. AB - OBJECTIVE: To investigate whether glycidly mechacrylate (GMA-DNA) adducts can be produced in various organs of rat in vivo. METHODS: Rats divided into 5 groups were orally administrated with mutagen, GMA 31.25, 62.5, 125, 250 mg/kg respectively for 14 days. DNA adducts produced in liver, kidney, white blood cells and testis had been analyzed by nuclease P1 mediated 32P-postlabelling method. RESULTS: Several GMA-DNA adducts were formed in various organs (in white blood cells, 4 types, liver and kidney, 3 types and testis 1 type). The amount of GMA-DNA adducts increased with GMA dosage within 0-125 mg/kg dosages, degree of the overall level of GMA-DNA adducts in various organs were kidney > liver > white blood cells > testis. N3-methacrylate-2-hdroxypropyl-deoxycytidine monophosphate was found in kidney, liver and white blood cells. CONCLUSIONS: GMA with electrophilic group could react with negatively charged centers on DNA and form GMA-DNA adducts. PMID- 12567492 TI - [The effect of interleukin-1 beta on cell growth of normal mouse pituitary cells and ACTH secreting adenoma]. AB - OBJECTIVE: To investigate whether IL-1 beta could affect the cell proliferation of normal pituitary cells and ACTH secreting adenoma cells. METHODS: Normal mouse pituitary cells were cultured in mininum essential medium-D-valine medium, a cultures virtually free of fibroblasts. AtT20 cells were also cultured as an ACTH secreting adenoma model. MTT method was used to study 10(-9) mol/L-10(-13) mol/L IL-1 beta effect on both normal and adenoma cells growth. RESULTS: (1) In the primate cells culture of normal mouse pituitary, IL-1 beta failed to influence cell growth during a period of 72 hours (n = 9, P > 0.05). This finding can not confirm that IL-1 beta has no effect on pituitary cell growth, because of the variety of cell types in pituitary. (2) In the AtT20 cells, 10(-9) mol/L-10(-13) mol/L IL-1 beta stimulated cell proliferation significantly after 48 and 72 hours culturing respectively (n = 9, P < 0.05, P < 0.01). CONCLUSIONS: These results suggested that IL-1 beta might be partially in involved in modulating pituitary adenoma cells growth. PMID- 12567494 TI - [Studies on the plant hormones produced by 5 species of endophytic fungi isolated from medicinal plants (Orchidacea)]. AB - OBJECTIVE: To study the plant hormones produced by 5 species of endophytic fungi isolated from medicinal plants and to illustrate the mechanism on endophytic fungi stimulating the growth of plants. METHODS: Extracting plant hormones from mycelia and its culture solution with organic solvent, and detecting them by HPLC. RESULTS: One or more plant hormones [GA3 (Gibberellin), IAA (Indoleacetic acid), ABA (Abscisic acid), Z (Zeatin), ZR (Zeatin riboside)] were detected from the mycelia and its culture solution. CONCLUSIONS: The plant hormones produced by the endophytic fungi are important materials that may be used to reveal the mechanism of endophytic fungi stimulating the growth of medicinal plants (Orchidacea). PMID- 12567495 TI - [The relation of fetal growth and the biological risk factors of coronary heart disease]. AB - OBJECTIVE: To examine the relationship between fetal growth and the biological risk factors of coronary heart disease (CHD) in adult life in China. METHODS: Follow up stuty of 628 men and women who were singleton born during 1948-1954 whose birth size were measured in detail. RESULTS: Decreasing birth weight increased the probability of myocardial ischaemic symptom related to CHD (beta = 0.0018, P < 0.01). The Odds ratio (OR) related to hypertension increased with decreasing birth weight and ponderal index (PI, kg/m3). The OR were 5.9 in people whose PI were 24 kg/m3 or less compare to people whose PI were more than 28 kg/m3. The OR related to impaired glucose tolerance(IGT) also increased with decreasing birth weight and PI. Insulin resistance, represented by fast insulin, rose with decreasing birth weight and head circumference(for trend, all P < 0.01). Serum TG concentration significantly rose with decreasing birth weight, PI and head circumference (for trend, P < 0.01, P < 0.01, P < 0.05), while HDL-C rose with increasing birth weight and head circumference (for trend, P < 0.05, P < 0.01). All above mentioned relations between birth size and the level of risk factors during adulthood were independent of gestation and adult lifestyle. CONCLUSIONS: In China, low birth measurements which represent the fetal growth retardation might contribute to the present of biological risk factors of coronary heart disease during adulthood, such as hypertension, diabetes, insulin resistance, high serum triglyceride and low HDL-C. PMID- 12567493 TI - [Clinical significance of detecting telomerase activity in ascites cells from patients with ovarian carcinoma]. AB - OBJECTIVE: The purpose was to investigate the clinical significance of telomerase activity for tumor diagnosis in ascites cells from patients with ovarian carcinoma. METHODS: Thirty ascites specimens from patients with ovarian tumor were examined for telomerase activity by PCR-based TRAP (telomeric repeat amplification protocol) method, and the result of detection was compared with the cytological diagnosis. RESULTS: Telomerase activity was detected in ascites cells from patients with ovarian tumor in 19 of the 20 cytologic-possitive specimens and in 3 of the 7 cytologic-negative specimens and in all the 3 cytologic suspicious specimens. The positive rate in 30 ascites samples by cytological examination and by the detection of telomerase activity was 66.67% (20/30), and 83.33% (25/30), respectively. CONCLUSIONS: The result suggested that measuring telomerase activity in ascites cells would be helpful to cytological examination in diagnosis of patients with ovarian carcinoma. PMID- 12567496 TI - [Study on the posterior vitreous detachment in patients with high myopia]. AB - OBJECTIVE: To access the development of posterior vitreous detachment(PVD) related to age, axial length of the eye, vitreous changes as well as the myopic chorioretinal manifestation in patients with high myopia. METHODS: The vitreous status of 61 consecutive patients with high myopia(> or = -8.0 D and axial length > or = 26.0 mm) 110 eyes and 68 controls (< or = -3.0 D) 119 eyes were examined with biomicroscopic slit lamp and B-scan ultrasound. The myopic chorioretinal changes at the posterior pole were examined by indirect ophthalmoscope. The axial length of the eye was measured by A-scan ultrasound. The incidence of PVD was evaluated by Chi-square test. RESULTS: The incidence of PVD in patients with high myopia increased with age. Its overall incidence was significantly higher than that in the controls (78.2% vs 31.1%, P < 0.001). In addition, the onset of PVD in patients with high myopia occurred much earlier in age than that in the control group. Axial length was an important factor responsible for the development of PVD in patients with high myopia. The incidence of PVD and age were highly correlated (r = 0.9997, P < 0.05). Retinal detachment occurred in 5 eyes in the patients with high myopia. CONCLUSIONS: The results in the present study suggested that the incidence of PVD in patients with high myopia, increased with axial length of the eye and age. Vitreous changes are characteristic findings associated with high myopia, which was also involved in the development of PVD. PMID- 12567497 TI - [Effect of space flight on ultrastructure of chloroplast in Agastache rugosa]. AB - OBJECTIVE: To substantiate the effects of space condition on medicinal plants, the seeds of Agastache rugosa were set up in retrievable satellites. METHODS: After returning to earth, the ultrastructural changes of chloroplasts were examined by means of electron microscope. RESULTS: Vesicles could be seen in the chloroplasts in the cells of earth controlled group, it was few and could not be found in the chloroplasts in the cells of weightless group and hit group. Some chloroplasts were degrading in the cells of weightless group, but this phenomenon was not observed in the cells of the other two groups. CONCLUSIONS: These results showed that space environment had some effect on the ultrastructure of chloroplasts in Agastache rugosa. PMID- 12567498 TI - [Angiotensin II regulates the expression of collagens in rat heart]. AB - OBJECTIVE: To testify the effect of angiotensin (Agn) II on the regulation and expression of collagen I, III genes in rat heart as well as the influence of DMP811 (a novel nonpeptide selective angiotension type 1 antagonist) on the effect of Ang II. METHODS: Six-week-old male SD rats with osmotic minipump subcutaneously containing either Ang II or saline were used. (1) Saline infused(control) group (n = 6), (2) Ang II-infused group (n = 6). (3) Ang II infused and DMP811-treated (3 mg.kg-1.d-1, by gavage) group (n = 6). Ang II (200 ng.kg-1.min-1) or saline was continuously infused for 1 week. After extraction of cardiac total RNA, the levels of collagen I, III mRNA and proteins were measured with RT-PCR and Sirius red staining respectively. RESULTS: Cardiac weight, cardiac weight/body weight radio, the mRNA levels of collagen I, III were increased by (4.7 +/- 0.4)%, (4.9 +/- 0.9)%, (22.0 +/- 4.7)% and (20.6 +/- 4.9)% respectively, in group 2 compared with those in group 1. The analysis of collagen on Sirius red-stained sections showed that Ang II increased the deposits of both collagen I and III. The above changes caused by Ang II could be inhibited fully by DMP811. CONCLUSIONS: Ang II up-regulated the expressions of collagen I, III via angiotension type 1 receptor. PMID- 12567499 TI - [Plasma branched-chain ketoacids metabolism in patients with chronic renal failure after oral administration of ketosteril]. AB - OBJECTIVE: To investigate the metabolism of branched-chain ketoacids (BCKA) in patients with chronic renal failure (CRF) and healthy control after an oral dose of ketosteril. METHODS: To determine plasma concentration of BCKA by gas chromatography/mass spectrography(GC/MS). RESULTS: Fasting plasma concentration of ketoisocaproate (KICA) and ketomethylvalerate(KMVA) in patients with CRF were lower than that of healthy control; The Cmax of KICA, KMVA and ketoisovalerate (KIVA) were (67.7 +/- 12.9), (57.8 +/- 10.3), (30.3 +/- 5.3) mumol/L; Tmax were (43.8 +/- 5.7), (58.1 +/- 4.6), (43.8 +/- 4.5) min; T1/2 were (108.5 +/- 12), (105 +/- 16.6), (116.8 +/- 18) min respectively. CONCLUSIONS: Absorption of BCKA in ketosteril had no significant difference between CRF patients and healthy controls; ketosteril could be used to treat CRF patients effectively. PMID- 12567500 TI - [Mode of action of sildenafil]. AB - Sildenafil is the first oral therapeutic agent for erectile dysfunction. Sildenafil is a selective inhibitor of cGMP-specific phosphodiesterase (PDE-5). Penile erection involves relaxation of the corpus cavernosum, an event mediated by NO and cGMP. The biological actions of cGMP are terminated by phosphodiesterase enzymes and PDE-5 is the major cGMP metabolising enzyme in this tissue. Sildenafil is relatively safe compared to erection injectables because it does not relax on isolated human corpus cavernosum, and does not cause priapism. Due to the tendency of abuse of sildenafil, its adverse cardiovascular associations with myocardial infaraction, ventricular arrhythmia and hypertension need to be alerted. PMID- 12567501 TI - [Observation of phagocytosis and cell junction of rabbit IPE in vitro]. AB - PURPOSE: To study the phagocytosis and cell junctions of iris pigment epithelium(IPE) in vitro. METHODS: Chicken red blood cells were added to culture medium of IPE, the numbers of IPE phagocyting chick red blood cells were counted; cell junctions were observed with transmission electron microscope. RESULTS: The phagocytotic rates of IPE and retinal pigment epithelium (RPE) were 5.4% and 8.8% respectively; there were various tight junctions in IPE. CONCLUSION: IPE possessd phagocytosis and barrier functions in vitro. PMID- 12567502 TI - [Amplifying variable region gene of light chain of monoclonal antibody against human retinoblastoma by PCR]. AB - OBJECTIVE: To acquire the variable region gene of light chain of monoclonal antibody against human retinoblastoma. METHODS: Total RNA were extracted from hybridoma cells secreting specific monoclonal antibody(McAb) against human retinoblastoma(RB), then transcripted reversely into cDNA with random primers. The variable region of the light chain(VL) gene fragments were ampliflied using polymerase chain reaction(PCR) method. Agrose gel electrophoresis was confirmed. RESULTS: 1.5% agrose gel electrophoresis indicated that VL gene was about 340 base pairs. CONCLUSION: The light chain variable region gene of the McAb against human RB was amplified successfully, which lays a good basis for construction of a recombinant antibody. PMID- 12567503 TI - [Tissue interaction with bioglass ceramic implanted in the rabbit cornea]. AB - PURPOSE: To evaluate the biocompatibility and 'healing' of Bioglass Ceramic(BGC) in the cornea. METHODS: BGC dicks (8 mm in diameter, 0.5 mm thick, 20-70 microns pore diameter, 37-62% porosity, 7.8 mm radius of curvature) were implanted in 11 New Zealand albino rabbits after a free-hand intralamellar dissection. The eyes were examined by slit-lamp biomicroscope up to seven months postoperatively. The corneas were removed at seven month (one eye at eight month), then were examined by light microscope. RESULTS: Five of 11 dicks of BGC were extruded. All implant of BGC with 51-62% porosity were broken and then extruded. A large part of implants led to markedly edematous with severe degrees of corneal neovascularization within one month postoperatively. The corneal lamella before implants became not clear in all eyes after two months. Lipid deposits occurred in four eyes. It is difficulty to observe the 'healing' process of BGC in the cornea because the BGC is too hard to be cut in spite of being soft by 4% hydrochloric acid for four to ten days before cutting. CONCLUSION: Our findings suggest that BGC with above parameters is not suitable as peripheral keratoprosthetic materials because of its rough, hard, thick and breakable. PMID- 12567504 TI - [The effect of frozen storage for amniotic membrane ultrastructure]. AB - OBJECTIVE: To observe the effect of frozen storage for amniotic membrane ultrastructure, and provide the experimental evidence for clinical use. METHODS: The amniotic membranes of fresh-obtained and -80 degrees C frozen storage 30 d, 60 d, 90 d and 180 d were examined with transmission electron microscope, and the ultrastructural changes were observed. RESULTS: The epithelium in the fresh obtained amniotic membrane was observed as intact structure, abundant organelles in cytoplasm and intercellular contact with desmosome. A great quantity of collagen micro-fibrils were found in the stroma, and the light and shade striation were observed in the collagen micro-fibrils of fresh-obtained amniotic membrane. The epithelium degenerated and died gradually as the increase of frozen storage time. At the frozen storage of 90 days, the epithelial chromatin dissolved, organelles degenerated and mitochondrion vacuolated. At the frozen storage of 30 days and 60 days, the pattern of collagen micro-fibrils in stoma was similar to that in fresh-obtained amniotic membrane; At the frozen storage of 90 days, some of collagen micro-fibrils in stoma showed intumescence. At the frozen storage of 180 days, the number of collagen micro-fibrils in stoma decreased, and some of them showed intumescence and dissolvent. CONCLUSION: Frozen storage can result in amniotic membrane epithelium death and collagen micro-fibrils intumescence and dissolvent. PMID- 12567505 TI - [Expression significance of P21WAF1/CIP1 protein in eye-lid primary malignant tumor]. AB - PURPOSE: To study the expression of P21waf1/cip1 protein in eye-lip primary malignant tumor (E-LPMT). METHODS: Immunohistochemical streptavidin biotin peroxidase complex(SP) method was used to stain section of E-LPMT. RESULTS: In 61 cases of E-LPMT, positive expression of P21waf1/cip1 protein was found locating in the nuclei. In Cell carcinoma, Squamous and Adenocarcinomas, the positive rate of P21waf1/cip1 protein expression were 36.0%, 27.8% and 22.2% respectively. The positive rate of P21waf1/cip1 protein expression in well-differentiated Cell carcinomas or Adenocarcinomas was higher than in poor-differentiated ones (P < 0.01). CONCLUSION: P21waf1/cip1 protein expresses in various type of E-IPMT and its positive expression in Cell carainomas and Adenocarcinomas are related to their different degrees. PMID- 12567506 TI - [Effects of angiotensin II on the 3H-TdR incorporation and synthesis of collagen in cultured bovine trabecular meshwork cells]. AB - OBJECTIVE: To investigate the effects of angiotensin II (Ang II) on the cell proliferation and collagen synthesis of bovine trabecular meshwork (TM) cells in vitro. It may probe into the mechanism of primary open-angle glaucoma(POAG). METHODS: (1) The bovine trabecular meshwork cells were cultured and identified by immunohistochemistry method(neuronal specific enolas, NSE, factor related antigan). The growing characteristics and morphological feature of cultured primary and passaged cells were observed by invered microscope and eletron microscope; (2) Ang II(1 x 10(-7) mol.L-1 and 1 x 10(-8) mol.L-1) and angiotensin receptor type I (AT1) antagonist (losartan) were incubated with cultured TM cells. The cellular proliferation was measured by 3H-thymidine(3H-TdR) incorporation assay and collagen synthesis was indirectly professed though detects the hydroxyproline of medium with chemistry methods. RESULTS: The cells of bovine trabecular meshwork were cultured successfully. Most of them were epithelia type. Ang II increased the TM cells uptake of 3H-TdR and inhibited by losartan partially. Meanwhile the hydroxyproline of medium was increased correspondingly. CONCLUSION: Establishing the method of culturing bovine trabecular meshwork cells is an important method for researching the characteristics of TM cells. Ang II can induce the cell proliferation of bovine TM cells and increase the synthesis of collagen in vitro. The AT1 antagonist may inhibit this cell proliferation effect. PMID- 12567507 TI - [Effects of pressure induced retinal ischemia on ERG in rabbit]. AB - OBJECTIVE: To observe the effects of pressure induced retinal ischemia on electroretinogram(ERG) in rabbit. METHODS: Retinal ischemia was induced in rabbits by increasing intraocular pressure at 30 mmHg, 60 mmHg, 90 mmHg, 120 mmHg for 45 minutes, and retinal function was monitored by eletroretinography. RESULTS: There was no difference on ERG before or after the experiment both in 30 mmHg group and control one. In 60 mmHg pressure induced ischemia eyes, the amplitudes of the b-wave and OPs wave reduced significantly. Four hours after reperfusion, they were totally recovered. After an ischemic insult of 90 mmHg or 120 mmHg for 45 minutes, there was no response of ERG. Four hours later, the amplitudes of the b-wave and OPs wave were 66.912 +/- 20.157 and 16.423 +/- 3.965 the former, 38.852 +/- 23.438 and 8.610 +/- 12.090 the latter, respectively. CONCLUSION: These results suggest that higher intraocular pressure causes more severe retina ischemic damage, and less recovery ability. PMID- 12567508 TI - [The measurement of multifocal electroretinography]. AB - PURPOSE: Discussing the testing procedure of multifocal electroretinography(ERG). METHODS: The testing procedure of 131 multifocal ERG records was reviewed. RESULTS: Some variable factors existed in the measurement of multifocal ERG. These factors included the subject's co-operation, the mounting of electrode, the fixation of tested eye and adjustment of focus. The good records of multifocal ERG depend upon the skill of the operator. CONCLUSION: The testing procedure of multifocal ERG must be standardized so as to ensure the reliability and repeatability of the results. PMID- 12567509 TI - [The effects of perfluorodecalin on the anterior chamber angle and intraocular pressure in rabbits]. AB - OBJECTIVE: To observe the effects of purfluorodecalin on the anterior chamber (AC) angle and intraocular pressure (IOP) in rabbits. METHODS: 0.05 ml perfluorodecalin was injected into right AC of 17 white rabbits and the same does of balanced salt solusion(BSS) was given to the controll group (6 rabbits). The IOP was measured before and 1, 2, 3, 4, 8, 12, 16 weeks after the injection. The treatment rabbits were sacrificed on the week of 1, 2, 4, 8 and 16 postoperatively. The eyes were enucleated and the histological examination was performed. RESULTS: There's no statistic IOP difference in the controll group before and after the injection of BSS. In treatment group, the IOP is similar to the preoperation level (11.62 +/- 6.35 mmHg) on the 7th postoperative day (10.82 +/- 3.17 mmHg, P < 0.05). A increment of IOP was measured at the week of 2 to 4, 8, 12, and 16 postoperatively (P < 0.01). Perfluorodecalin, having a 'fish eggs' or bubbles appearance, was noted in the inferior AC. Histopathologically, transparent single or fused vesicles and macrophages were accumulated at the AC angler in treatment rabbits. Some lymphocytes and neutrophils were aslo noted here. Trabeculae degenerated and intertrabecular space increased. CONCLUSIONS: The anterior chamber injection of perfluorodeclin could lead to a longstanding increase of IOP. Inflammatory reaction and trabeculae degeneration were seen in the anterior chamber angler. PMID- 12567510 TI - [Analysis of MRI for tumor of the sellar region]. AB - PURPOSE: To introduce the characteristics of magnetic resonance imaging(MRI) for the tumors of the sellar region. METHODS: A retrospective analysis of 86 cases with the tumors of the sellar region was carried out. RESULTS: The pituitary tumor including the pituitary micro-adenoma and pituitary macro-adenoma was a common tumor of the sellar region. The pituitary micro-adenoma usually does not affect visual function; but the pituitary macro-adenoma often damages the optic pathway and reduces the visual function. The rest of the sellar region appears special feature on MRI. CONCLUSION: MRI is a useful method in diagnosing the tumors of the sellar region. PMID- 12567511 TI - [The clinical research of phaco techniqiue and IOL implantation in children cataract surgery]. AB - OBJECTIVE: To investigate the clinical application of phaco technique with amblyopia treatment in children cataract. METHODS: The cataract surgery and intraocular lens (IOL) implantation were done in 21 cases(36 eyes) of children who were from three to 12 years old with amblyopia treatment postoperation. RESULTS: The postoperative corrected visual acuity was more than 0.5 on 12 eyes (33%), from 0.1 to 0.4 on 13 eyes (36%), less than 0.1 on Three eyes (8.3%), incorporation on eight eyes (22%). Strabismus was found on one eye with nystagmus on one eye and central corneal perforation on two eyes. In the follow-up, there were two eyes with platycoria and three eyes posterior capsular opacity with Erbium: Yttium Alminum-Garnet Laser(YAG) laser capsulectomy, especially on the patients with traumatic cataract. CONCLUSION: The chilren cataract surgery with the application of phaco technique and amblipia treatment postoperation were more excellent than those with the traditonal surgical methods. We should attach importance to amblipia treatment after the operation. PMID- 12567512 TI - [The management of posterior capsular in cataract operations of high myopia]. AB - OBJECTIVE: To study the management of posterior capsular in cataract operation of high myopia in order to avoid posterior capsular opacity(PCO). METHODS: Twenty two cases(30 eyes) of high myopia with an average axial length of 31.62 mm were analyzed. Cataract phacoemulsification were performed in all cases, among which 12 eyes were operated with continuous curvilinear capsulorhexis (group I) of posteror capsular and 18 eyes were operated with capsulorectomy combined with anterior segment vitrectomy (group II). No artificial lenses were implanted in all cases. RESULTS: The post-operative naked vision were improved in both groups. Group I: The VA > or = 0.1 in all 12 eyes(100%), > or = 0.3 in seven eyes (58.3%) and no complication happened. Group II: The VA > or = 0.1 in 16 eyes(88.9%), > or = 0.3 in nine eyes (50%), and one eye with hemmorhage choroid detachment in the operation and three eyes with retina tear after the operation. CONCLUSION: Continuous curvilinear capsulorhexis of posteror capsular in treating PCO of high myopia with cataract is a safe effective and economic way compared with capsulorectomy combined with anterior segment vitrectomy. PMID- 12567513 TI - [The ocular hypotensive effect and safety of 0.2% brimonidine]. AB - PURPOSE: To study the ocular hypotensive efficacy and safety of 0.2% Brimonidine when used in chinese people. METHODS: Using randominzed parallel group control study, 0.2% Brimonidine twice a day or 1% Carteolol twice a day was applied in patients with primary open angle glaucoma or ocular hypertension for three months. RESULTS: The results showed that 0.2% Brimonidine and 1% Carteolol both have good ocular hypotensive efficacy. There was no significant difference between them. 0.2% Brimonidine did not produce significant changes in pupil size and heart rate. Change in blood pressure was not clinically significant. Six patients with 0.2% Brimonidine had drowsiness. Two patients had dry mouth. One patient had ocular burning. CONCLUSION: Our results suggested that 0.2% Brimonidine would be a stable and effective antiglaucoma medicine with good tolerance. Thus, 0.2% Brimonidine should be the main choice of medical therapy for primary open angle glaucoma and ocular hepertension. PMID- 12567514 TI - [Microtrebeculectomy in glaucoma]. AB - OBJECT: To study the clinical effectiveness of microtrabeculectomy. METHODS: Of 80 patients (80 eyes) with various types of glaucoma, 40 (40 eyes) underwent microtrabeculectomy. The scleral flap measures 2 mm x 2 mm. The rectangular resection of trabeculum measures 1 mm x 1 mm. And 40(40 eyes) underwent standard trabeculectomy with the scleral flap measuring 3 mm x 4 mm and the resection of trebeculum measuring 1.0 mm x 3.0 mm. Vision acuity, intraocular pressure (IOP) measurements, slit lamp examinations and opthalmicroscope examinations were performed before surgery and after surgery at one, two, eight and 15 days, and one, two, three, six, nine, 12, and 18 months. RESULTS: The mean preoperative IOP was 44.11 +/- 14.86 mmHg(1 mmHg = 0.133 kPa) for microtrabeculectomy and 46.45 +/ 11.27 mmHg for standard trabeculectomy. The mean postoperative IOP and operative complications were similar between the two groups. The mean postoperative IOP was 12.94 +/- 4.13 mmHg for the group with microtrabeculectomy and 13.51 +/- 3.56 mmHg for the group with standard trabeculectomy at the end of follow-up (12-18 months). The incidence of shallow anterior chamber was 16% (6/40) in micro group, while it was 2.6% (1/40) in the other group. There was no significant difference between the two groups(X2 = 1.59 < 3.84, P > 0.05). CONCLUSION: Microtrabeculectomy appears to provide reasonable control of IOP. PMID- 12567515 TI - [Vitrectomy and photocoagulation for treatment of proliferative diabetic retinopathy]. AB - PURPOSE: To evaluate the efficacy of vitreous surgery and photocoagulation for treatment of a series of patients with proliferative diabetic retinopathy(PDR). METHODS: A consecutive series of PDR treated with vitrectomy and photocoagulation during February 1997 to May 2000 was retrospectively analyzed. RESULTS: The series included 79 cases (38 male and 41 female) and 104 eyes [oculus dexter(OD), 26; oculus sinister(OS), 28; oculus uterque(OU), 25] with diabetic history for 0.5-37 (8.5 +/- 6.5) years and eye symptoms for 15 days to 1.5 years (1.5 +/- 1.5 years). Retinal photocoagulation was performed in 30 eyes with only once in 26 eyes before vitrectomy. Surgical techniques included standard vitrectomy, neovascular membrane peeling and fragmentation, endo--diathermy and endolaser. Silicone oil was injected in 59 eyes with active bleeding, retinal detachment, or severe ischemia. Cataract extraction was done in 63 eyes, of which 27 eyes with intraocular lens implantation. Visual acuity was light perception(LP), hand movement(HM), count fingers(CF), and > or = 0.02 in 15, 38, 22, and 29 eyes, respectively before operation. Visual improvement achieved in 84 eyes (80.8%) with > or = 0.1 in 41 eyes (39.4%). Visual acuity remained unchanged or decreased in 20 eyes (20.2%). Neovascular glaucoma occurred in two eyes after surgery. CONCLUSIONS: Although the series of patients with PDR without sufficient retinal photocoagulation before operation, vitrectomy and endolaser improved the visual outcome in the majority of eyes. Silicone oil injected in eyes with severe ischemia, active bleeding or retinal detachment may provide chance for prompt laser coagulation after surgery in order to prevent progression of PDR. PMID- 12567516 TI - [A review of research actuality on age-related macular degeneration]. AB - A research actuality on Age-related Macular Degeneration(AMD) is reviewed, including epidemiology, pathogeny, mechanism, and the therapy of western or traditional chinese medicine. The pathogeny of AMD is linked with age, race, the history of heredity, smoking and environment. The mechanism of AMD is concerned with the metabolic disturbance of RPE and the chronic nonspecific inflammation. The main therapies for treating AMD include laser photocoagulation, photodynamic therapy, surgery, radiotherapy, drug and gene therapy. Using traditional chinese medicine to treat AMD is an exploratory direction which deserves to be mentioned due to the present situation of no effective treatments to AMD. PMID- 12567517 TI - [Clinical analysis of 184 eyes with perforating injury]. AB - OBJECTIVE: Try to understand the occurrence and the prognosis of eyes with perforating injury. METHODS: By using retrospective analysis on 184 eyes with perforating injury. RESULTS: Perforating eyes hold 9.5% of total impatient, mostly happened in working place, foreign body is the main cause, the age is mostly from ten to 60, centralize from 20 to 30. the complication includes leucoma, cataract, vitreous hemorrhage etc. After operation, 71 eyes had visual acuity < 0.05. 60.85% of the patients had visual acuity > or = 0.05 and 0.65% of the patients are not cooperative. CONCLUSION: The rate of blindness on eyes with perforating injury is high, it implies that perforating injury could be prevented and treated from the data we collected. PMID- 12567518 TI - [Blossom and growth of pangdahai at Hainan Island]. AB - Two species of Pangdahai--Scaphium lychnophorum Pierre and Scaphium wallichii Schoott & Endl. were introduced into Hainan Island in 1970s. Over a long-term observation and investigation at four different trial points, the results showed the main limited factor of the cultivation was typhoon; only in Nada area, northwest in Hainan Island, the upper two species grown well and bloomed after 14 16 years of planting, but not fruit. The analysis of the causes of the non blossom and fruitlessness have also been made in this article. PMID- 12567519 TI - [Investigation and identification of varieties of wulingzhi in Qinghai Province]. AB - This paper reports the investigation and identification of varieties of Wulingzhi used as drugs in Qinghai province. The results show Wulingzhi varieties from the province and the parts of northwest districts are different with the Wulinzhi recorded in China Pharmacopoeia, which has itself special varieties and identified standard, their originals are the excrements from Petaurista xanthotis, Ochotona erythrotis, O. curzoniae. O. daurica and O. causus. PMID- 12567520 TI - [Pharmacognostic identification on crude drug of Dendrobium denneanum]. AB - This paper reported the morphological and histological characteristics of Diequiao Shihu(the crude drug of Dendrobium denneanum Kerr), with linedrawings, and compared the TLC diagram with Jichai Shihu (the crude drug of Dendrobium nobile Lindl.). PMID- 12567521 TI - [Identification of Radix and Rhizoma Rhei by UV and TLC]. AB - This paper deals with the identification of radix and rhizoma of 24 species (varieties) of Genus Rheum by UV and TLC. This results provide authentic methods for the identification of Radix and Rhizoma Rhei. PMID- 12567522 TI - [Effects of different amount of ginger juice on prepared products of Rhizoma Pinelliae]. AB - Rhizoma Pinelliae were diged in Ginger juice of different propertion, the total alkaloid contents of these products were all higher than the raw Rhizoma Pinelliae, and related to the actions of anti-tumous cell. The contents of beta sitosterol of these products were related to the amount of Ginger juice which is added in Pinelliae Rhizoma, but only the content of beta-sitosterol of the Pinelliae product which is prepared by same amount of Ginger juice was slight higher than the raw Rhizoma Pinelliae. The stimulations of five kinds of products on mice have no obvious relation to the amounts of Ginger juice which were added in Rhizoma Pinelliae, but the stimulation of the Pinellia product which was prepared by same amount of Ginger juice was the lowest of all products on mice. Dependent on above experiment results, we suggest that Rhizoma Pinelliae processed by Ginger juice should be prepared by same amount of Ginger juice with Rhizoma Pinelliae. PMID- 12567523 TI - [Studies on the chemical constituents of soft coral Lemnalia bournei]. AB - The studies on the chemical constituents of soft coral Lemnalia bournei led to the isolation of 2-deoxy-12-oxo-lemnacarnol, 22,23-dihydrobrassicasterol, gorgosterol and batyl alcohol. Their structures were deduced on the comprehensive spectral analysis. The four compounds were first obtained from the soft coral. PMID- 12567524 TI - [Origin confirmation of a new natural product from Oldenlandia diffusa]. AB - A new natural product, 4,4'-dihydroxy-a-truxillic acid (I) was isolated from Oldenlandia diffusa (Willd) Roxb. I is a derivate of cyclobutane, which were obtained chemically by photochemical reaction. Our works confirmed that the compound is a plant metabolite, not a secondary product produced during its extraction and isolation. PMID- 12567525 TI - [Studies on the chemical constituents of Citrus reticulata]. AB - Four compounds were isolated from the pericarp of Citrus reticulata and three of them were identified as natsudaidai, nobiletin and 3,5,6,7,8,3',4' heptamethoxyflavone. Natsudaidai and 3,5,6,7,8,3',4'-heptamethoxyflavone were obtained from this plant for the first time. PMID- 12567526 TI - [Effect of eel oil capsule on mice memory]. AB - Step-down test, step-through test and water maze in mice were used to observe the effect of eel oil capsule on mice memory. The results indicated that during the administration period (30 days), the eel oil capsule treating groups with dosages of 0.129, 0.387, and 1.161 g/kg obviously improved the learning and memory abilities. Compared with the control test, the eel oil capsule promoted the memory and smart. PMID- 12567527 TI - [The further studies on the pharmacological actions of Bupleurum smithii var. parvifolium]. AB - Bupleurum smithii var. parvifolium grown widely in Ningxia, had a long history as a common medicinal herb. Its crude saponin had significant protective effect in hepatic damage caused by CCl4 in mice, its ether extract had significant analgesic effect. The ether extract also had significant immunological facilitation effect in mice, it could increase the thymus gland weight and the serum IgG content. These results were similar to or better than that of B. chinense and indicated that B. smithii var. parvifolium could be used as B. chinense. PMID- 12567528 TI - [Study on quality evaluation of Chinese traditional patent medicine with chemical patterm recognition (I)]. AB - In this paper, the quality of Wuji Baifeng pills was comprehensively evaluated by a method of chemical pattern recognition, all the samples were divided into three grades (good, middling and poor). This method can be popularized and applied to evaluate quality of other Chinese traditional patent medicines. PMID- 12567529 TI - [Screening and identification of egg mimotope of Schistosoma japonicum]. AB - OBJECTIVE: To explore egg mimotope of Schistosoma japonicum that can be used in the development of diagnostic reagents for schistosomiasis. METHODS: By performing three rounds of biopanning in the affinity selection before picking out single clones for identification, target specific phages were effectively enriched from a random 15-peptides phage library with an immobilized mAb 6B12 which is specific to egg antigen of S. japonicum as a bait. By using ELISA, competitive ELISA with natural egg antigen as competitor, and Western blotting, 13 phage clones with high affinity and specificity to 6B12 were obtained and amplified from 400 single clones. RESULTS: DNA sequencing revealed that all the 13 selected single clones were identical in displaying gene sequence. Serum samples were tested by ELISA for the presence of IgG antibodies to the phages with mimic epitope, and showed that there was a significant difference between healthy volunteers and schistosomiasis patients. CONCLUSION: It is possible that the phage display peptide antigen, egg mimotope of S. japonicum may be a replacement of natural egg antigen for the diagnosis of schistosomiasis. PMID- 12567530 TI - [A case of hypodermiasis with empyema]. PMID- 12567532 TI - [Study on molecular phylogeny of Schistosoma sinensium based on mitochondrial genes]. AB - OBJECTIVE: To determine the phylogenetic position of Schistosoma sinensium in the genus Schistosoma using mitochondrial cytochrome C oxidase 1 (CO1) and NADH dehydrogenase 1(ND1) as molecular markers. METHODS: The genomic DNA of adult worms were extracted by the GNT-K method. The target regions were amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid Zero-Blunt. Recombinant plasmids were amplified in E. coli, extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of related schistosomes were retrieved from GenBank and aligned with our data in the sequence editor ESEE. Gene trees were constructed in PHYLIP and MEGA using both maximum parsimony and neighbor-joining methods. For parsimony analysis, all characters were treated as unordered and with equal weights. At least 3,000 cycles of bootstrapping were carried out. For analysis in MEGA, all gap columns were deleted. The third position of codon was included. RESULTS: The nucleotide and amino acid sequences of CO1 and ND1 of S. sinensium were obtained. CONCLUSION: The phylogenetic trees from these molecular data suggested that S. sinensium belongs to the Asian schistosome group, and the results coincided with the previous rDNA (ITS2 & LSU) analysis results. PMID- 12567531 TI - [Spatial autocorrelation analysis on schistosomiasis cases and Oncomelania snails in three provinces of the lower reach of Yangtze River]. AB - OBJECTIVE: To explore the spatial distribution of schistosomiasis cases and Oncomelania snails in the provinces of Jiangsu, Anhui and Jiangxi with the assistance of geographic information systems (GIS). METHODS: Epidemiological data of schistosomiasis in recent 20 years were collected, and the relevant GIS databases were established. The spatial autocorrelation analysis was carried out with ArcView 3. x, S-Plus, and Spatial Statistics software. RESULTS: Except for schistosomiasis cases in Jiangsu, the spatial distribution of schistosomiasis cases and Oncomelania snails in the three provinces were fitted to the spatial autocorrelation at a certain degree. Generally the autocorrelation coefficients (Moran's I) in Oncomelania snails were higher than that in schistosomiasis cases. CONCLUSION: Spatial autocorrelation analysis can be used to estimate the value of one variable in different spatial position, and it is a good way to find out the spatial cluster distribution in different stages. PMID- 12567533 TI - [Screening of stage-specific expression genes of Spirometra erinacei-europaei plerocercoid by mRNA differential display technique]. AB - OBJECTIVE: To screen stage-specific expression genes of plerocercoid of Spirometra erinacei-europaei. METHODS: RNA was extracted by the acid guanidinium thiocyanate-phenol-chloroform from plerocercoid and adult worm of Spirometra erinacei-europaei. Contaminated DNA in the RNA was digested by RNase-free DNase. cDNA was synthesized by using T12MA, T12MC, T12MG and T12MT primers, and PCR was then done using the same T12MN and one random primers. The PCR products were fractionated on 8% denatured polyacrylamide gel, differential bands of plerocercoid found in the gel were cut out, amplified by PCR and sequenced after the gel was dried out and autoradiographed. Northern hybridization was conducted to identify the stage-specific expression genes. RESULTS: Eleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization after they were amplified by PCR. The fragments 1 and 2 were confirmed to express specifically in plerocercoid by Northern hybridization, but the fragment 3 was expressed in both plerocercoid and adult worm. When the 3 gene fragments were homologically analyzed in GenBank, the sequence which was homologous with the fragments 1 and 2 was not found, but the fragment 3 had high homology with many kinds of 28S rRNA. CONCLUSION: The gene expression of plerocercoid was different from that of adult worm probably because they live in different hosts. Two kinds of different gene fragments in plerocercoid were identified by mRNA differential display technique. PMID- 12567534 TI - [Prevention and control of intestinal nematode infection in Shanghai]. AB - OBJECTIVE: To evaluate the current status of intestinal nematode infection in Shanghai and make recommendations for further control activities. METHODS: Retrospective review of the control program was made based on the change of nematode prevalence in Shanghai area since the 1950s, and challenges under the current situation were analysed. RESULTS: The intervention measures included chemotherapy, health education, sanitary disposal of human excreta and safe water supply. With the control strategy, socio-economical development and the change of farming patterns, the prevalence of intestinal nematode infection in population was reduced by 88.5%, from 62.6% in 1955 to 8.2% in 2000. Among them, ascaris infection decreased by 89.0%, from 52.1% to 6.3%, hookworm infection decreased by 99.0%, from 12.9% to 0.1%, but pinworm infection was still as high as 18.9% in 1999. CONCLUSION: Prevalence of nematode infection in Shanghai was considerably reduced. To match it with the current socio-economical development in the Municipality, however, more needs to be done. Sustainable control activities and surveillance are recommended. PMID- 12567535 TI - [A case-control study on risk factors for advanced schistosomiasis japonica]. AB - OBJECTIVE: To investigate risk factors for the development of advanced schistosomiasis. METHODS: The case-control study was designed with a match of 1:1 and 1:2. Healthy persons and chronic schistosomiasis patients were used as control. Each group was composed of 213 cases. Items investigated included: history of schistosomiasis and treatment, exposure to the contaminated water, social-economical condition and nutritional status, other concomitant diseases. Cellular and humoral immunity, HBVMs and ABO blood groups were also detected. Statistical analysis was performed by 1:1 and 1:2 matched single and stepwise conditional logistic regression analysis with SAS software. RESULTS: By stepwise conditional logistic regression analysis, it was revealed that number of schistosomiasis examination (OR = 1.168-1.311), interval from first infection to last treatment(OR = 1.142), interval from first infection to this investigation (OR = 1.089), HBsAg+ (OR = 4.683-10.759), HBcAb+ (OR = 2.873), HBsAg+ + HBeAb+ + HBcAb+ (OR = 7.64) were risk factors of developing advanced schistosomiasis. The average living space and cellular immunity were lower in advanced schistosomiasis patients than others. No association was found between advanced schistosomiasis and ABO blood groups. CONCLUSION: The development of advanced schistosomiasis is associated with repeated infections, and delayed or incomplete treatment. Combined infection with hepatitis B, poor socio-economic conditions and impaired cellular immunity may increase the pathogenetic risk of schistosomiasis. PMID- 12567537 TI - [A death due to misdiagnosis of cerebral falciparum malaria]. PMID- 12567536 TI - [Epidemiological study on taeniasis and cysticercosis in Shandong Province]. AB - OBJECTIVE: To investigate current epidemiological status of Taenia solium infection and cysticercosis in Shandong Province. METHODS: By house to house interview in the field, fecal examination and serological tests, the prevalence data of taeniasis and cysticercosis in human and pig population, and epidemiological factors in relation to the diseases were carried out. RESULTS: The prevalence of Taenia infection and cysticercosis in human population were 0.048% and 0.057% respectively. The sero-positive rate of the specific IgG4 of anti-Cysticercus was 1.91%. The prevalence differed among occupations and from region to region, but there was no significant difference between men and women, city and suburb. Unhealthy eating and unhygienic habits commonly practised among residents, 92.3% of the families have "latrines connected to pigsties". The surrounding sanitation and the disposal of fecal material were poor. The detection rate of porcine cysticercosis was 0.06%, and the positive rate of anti Cysticercus antibody in pigs was 1.99%. CONCLUSION: The prevalence of taeniasis and cysticercosis is decreasing. However, the endemic area is expanding and the epidemiological factors are still existing. Continued effort is therefore needed for sustain the control programme. PMID- 12567538 TI - [Sequence difference of ribosomal DNA second internal transcribed spacer in Anopheles minimus in different localities]. AB - OBJECTIVE: To compare the sequence difference of the ribosomal DNA second internal transcribed spacer (ITS2) in Anopheles minimus from Yunnan, Hainan, Guangxi of China and from Thailand. METHODS: Single mosquito's legs were digested to extract DNA; the rDNA-ITS2 region was amplified by specific primers and the PCR products were purified, sequenced and analysed. RESULTS: Two different kinds of ITS2 sequences, temporarily named as An. minimus A and An. minimus C, were found in this study. The length and GC contents of ITS2 region in species A and species C are 481 bp, 54.04% and 483 bp, 54.23%, respectively. Two species differed by 5.8% in the ITS2 sequences owing to 22 fixed nucleotide substitutions and indels. CONCLUSION: Two cryptic species, An. minimus A and An. minimus C, were found in the research localities. PMID- 12567539 TI - [Effect of malaria control measures in Rongshui County]. PMID- 12567540 TI - [Obtaining and construction of a novel gene from Schistosoma japonicum by expressed sequence tags]. AB - OBJECTIVE: To look for new candidate vaccine molecule and characterize new genes from Schistosoma japonicum. METHODS: New genes of S. japonicum were obtained through expressed sequence tags and analyzed on line. One interest cDNA was cloned into anti-sense eukaryotic expression vector. RESULTS: An interest cDNA JAYL0230 was obtained. Analysis showed that homology existed between its deduced amino acid sequence and defender against apoptotic death 1 of human and pig, etc. Its anti-sense cDNA was cloned into eukaryotic expression vector pEGFP-N3. CONCLUSION: The combination of expressed sequence tags and bioinformatics is an effective way to find novel genes of creature. Construction of anti-sense eukaryotic vector is helpful in studying gene's function. PMID- 12567541 TI - [The construction of monoclonal antibody cell line against Schistosoma japonicum recombinant protein 20.8 kDa antigen]. PMID- 12567542 TI - [Effect of MNNG on ornithine decarboxylase activity in cells from adult Schistosoma japonicum]. AB - OBJECTIVE: To observe the change of ornithine decarboxylase(ODC) activity in cells from adult Schistosoma japonicum after the cells were treated with N-Methyl N-Nitro-N-Nitrosoguanidine (MNNG). METHODS: The cells were treated with MNNG at a concentration of 3 micrograms/ml for 48 hours after the cells being incubated for one week. The cells were then cultured with RPMI-1640 containing 10% calf serum. ODC activity was detected with spectrophotography. RESULTS: ODC activity rose significantly in two to three weeks after the cells were treated with MNNG. CONCLUSION: There was ODC activity in cells from adult S. japonicum and MNNG has an effect to reinforce ODC activity in the cells. PMID- 12567543 TI - [Study on inducing an artemisinin-resistant line of Plasmodium berghei]. AB - OBJECTIVE: To induce a line of Plasmodium berghei with resistance to artemisinin. METHODS: The major methods included blood transmission from passage to passage and progressive increase of drug pressure. RESULTS: The resistant lines were developed by different protocols: (A) The initial dosage of artemisinin was 126.2 mg/kg which was increased by 60 mg/kg for the next passage and boosted by 126.2 mg/kg for every other passage. As developed to passage 60 and 76, the resistant index was 18.39:1 and 14.89:1 respectively, then decreased gradually. For passage 108, the dosage was 8,862.5 mg/kg, but the resistant index was only 10.49:1. (B) Using passage 66 from (A) as the source, a dosage of 4,000 mg/kg was given each week, the resistance of the passage 40 increased significantly with an index of 27.5:1. (C) Using passage 19 of (B) as the source, drug was administered at the dose of 2,000 mg/kg each week. The resistant index of passage 15 was 17.41:1. CONCLUSION: Line of P. berghei with medium level resistance to artemisinin was established. PMID- 12567544 TI - [Construction and characterization of the cDNA library from Schistosoma japonicum cercariae]. AB - OBJECTIVE: To construct a cDNA library from Schistosoma japonicum cercariae. METHODS: Total RNA was extracted from 500,000 S. japonicum cercariae and used to synthesize the double-stranded cDNA and cloned into the TriplEx2 vector using "SMART cDNA Construction Kit". RESULTS: The primary titer of the constructed cDNA library is 1.8 x 10(7) pfu/ml and the titer of amplified library is 2.5 x 10(10) pfu/ml. The average size of inserts is 1.075 kb. The recombinant efficiency is 94.4%. The full length cDNA of S. japonicum TPI and JF-2 genes was successfully amplified from the library. CONCLUSION: A cDNA library of S. japonicum cercariae has been constructed successfully. PMID- 12567545 TI - [Comparative effect of different media in in vitro cultivation of Trichomonas vaginalis]. AB - OBJECTIVE: To investigate the optimal condition for in vitro cultivation of Trichomonas vaginalis for obtaining a better harvest of T. vaginalis. METHODS: An isolate of T. vaginalis from clinical specimens was cultivated in three different media with initial inoculation of 9.0 x 10(4)/ml under pH 5.6. RESULTS: There was distinct difference after 96 h incubation in the cumulative harvest of T. vaginalis. The highest harvest was received in cysteine/liver/peptone/maltose medium, followed by the liver/peptone/maltose medium and soybean/liver/peptone/maltose medium. CONCLUSION: The cysteine/liver/peptone/maltose medium may be a suitable environment for in vitro multiplication of T. vaginalis. PMID- 12567546 TI - [A human case of Gnathostoma spinigerum phase three larve infection]. PMID- 12567547 TI - [Discussion about strategy and protocol of congenital toxoplasmosis screening]. PMID- 12567548 TI - [A misdiagnosed case of imported falciparum malaria]. PMID- 12567549 TI - [Neurotoxicity of artemisinin antimalarials and its mechanism]. PMID- 12567550 TI - [Opinion on hydatidosis control by immuno-prophylaxis]. PMID- 12567551 TI - [Sequencing of the glyceraldehyde-3-phosphate dehydrogenase gene from Plasmodium falciparum strain FCC1/HN]. PMID- 12567552 TI - [An outbreak of trichinosis in Jianshui County of Yunnan Province]. PMID- 12567553 TI - [Experimental studies on the life period of AnopheleS hyrcanus sinensis of Guangxi]. PMID- 12567554 TI - [Clinical analysis on 21 cases of enterobiasis in nasal cavity]. PMID- 12567555 TI - [Schistosomiasis investigation at 4 villages of Yangjiayuan during a flood in 1999]. PMID- 12567556 TI - [A case of cerebral schistosomiasis misdiagnosed as brain tumor]. PMID- 12567557 TI - [Study on cellular immunity function in child patient with cerebral cysticercosis]. PMID- 12567558 TI - [Effectiveness of caring on chronic cases of filariasis in Zhejiang Province]. PMID- 12567559 TI - [A brief history of the early stage of malaria control in China]. PMID- 12567560 TI - Functional characterization of the 5' proximal flanking fragment of Plasmodium falciparum GBP130 gene. AB - OBJECTIVE: To identify the putative regulation elements with strength- and stage specificity in 5' proximal flanking sequence of P. falciparum GBP130 gene. METHODS: Plasmids containing different deletions of upstream of the GBP130 promoter were constructed. For strength-specific analysis, pGBPCAT delta 2, pGBP delta 2/400 and pGBP delta 2/800 were electroporated into ring-stage P. f. respectively, and the expression level of CAT in each plasmid was detected by liquid scintillation counts(LSC). For stage-specific analysis, transfectants with pGBP delta 2/400 and pGBP delta 2/800 were harvested at 5 hours post transfection(h), 15 h and 46 h respectively, and the CAT expression levels were detected. RESULTS: In strength-specific analysis, the expression level of CAT in pGBP delta 2/800 and pGBPCAT delta 2 was similar, down-regulated significantly in pGBP delta 2/400. The CAT level showed significant difference between pGBP delta 2/400 and the control. In stage-specific analysis, the CAT level of pGBP delta 2/400 was higher than that of pGBP delta 2/800 at the time point of 5 h, and lower at 15 h and 46 h. CONCLUSION: This strength-specific promoter activity was due to the difference of 5'UTR length: the longer the 5'UTR the higher the promoter strength, and two poly (dA:dT) tracts in the proximal sequence could enhance the promoter activity. The length of 5'UTR regulated the promoter activity in a stage-specific manner. The shorter 5'UTR was functional at ring stage, while the longer one prompted transcription at trophozoite and schizont stage. The functional role of poly (dA:dT) tracts in stage-specific regulation of GBP130 remains unclear. PMID- 12567561 TI - [Cloning of a species-specific gene fragment from Cryptosporidium parvum and the development of diagnostic PCR primers]. AB - OBJECTIVE: To develop a pair of diagnostic PCR primers for Cryptosporidium parvum. METHODS: A species-specific gene fragment of C. parvum was obtained through RAPD analysis. After the fragment was isolated, purified, cloned and sequenced, a pair of primers FF was designed and synthesised based on the sequence. With the primers, the anticipated fragment in size of 603 bp was amplified by PCR from 2 American strains and 4 Chinese strains of C. parvum. The samples of 35 rabbits feces and 55 human feces were detected by PCR with primers FF and 021, the latter was a species-specific diagnostic primer reported by Morgan. RESULTS: All six strains amplified by the primers FF showed same detection rate with 021. Sensitivity test indicated that DNA of 1 oocyst per gram of feces could be detected by the PCR. CONCLUSION: The primers FF showed high specificity and sensitivity, and can be used for diagnosing Crytosporidium parvum infection. PMID- 12567562 TI - [Construction and expression of an eukaryotic recombinant plasmid from a hybrid gene of antigen for Plasmodium falciparum]. AB - OBJECTIVE: To construct an eukaryotic recombinant plasmid with HGFSP, a hybrid gene encoding the antigen epitopes of MSA1, MSA2, RESA and CSP in different developing stages of Plasmodium falciparum (P. f.). METHODS: HGFSP was sub-cloned into an eukaryotic expression plasmid pcDNA3 from a prokaryotic recombinant plasmid pSK-HGFSP to construct the eukaryotic recombinant expression plasmid pc HGFSP. The identified recombinant was then transfected into HepG2 cells with liposome-mediated method. The G418-selected positive cell clones were tested to identify the immunogenicity of HGFSP-expressing antigen. RESULTS: It was evidenced that HGFSP was correctly inserted into pcDNA3 by restriction enzymes map analysis. HGFSP-expressing antigen-specific fluorescent response was observed in pc-HGFSP-transfected HepG2 cells. The results of SDS-PAGE and Western-blotting showed that there was a 23 kD protein band, which can be specifically recognized by anti-sera of HGFSP-expressing protein in pc-HGFSP-transfected HepG2 cell lysis. CONCLUSION: Pc-HGFSP, an eukaryotic recombinant plasmid encoding hybrid antigen epitopes of P. f., was constructed successfully and the antigenicity of pc-HGFSP-expressing protein was confirmed. PMID- 12567563 TI - [Study on genomic diversity among different isolates of Schistosoma japonicum in China]. AB - OBJECTIVE: To study genomic diversity among the isolates of Schistosoma japonicum in China. METHODS: Schistosome adults were collected from different endemic areas. Mitochondrial NADH dehydrogenase 1 (ND1) and cytochrome C oxidase 1 (CO1) gene fragments of the worms were amplified by PCR, cloned into plasmid, and finally sequenced. Program MEGA was used for sequence analysis. RESULTS: The ND1 and CO1 genes amplified from different geographic strains were 476 bp and 1,033 bp, respectively. There are two distinct haplotypes, type I and II, for both ND1 and CO1 nucleotide sequences. The differences between the two types were 4.0% and 3.4%. They are considered to be two different genetypes by the phylogenetic analysis. In individual mitochondrial gene, type I of ND1 was fixedly accompanied with type I of CO1 gene, and type II of ND1 with type II of CO1. CONCLUSION: There are two different genetypes of ND1 and CO1 genes of S. japonicum in China. PMID- 12567564 TI - [Effect of treating slides on examining hookworm eggs by brine floatation technique]. PMID- 12567565 TI - In vitro antimalarial effect of daphnetin relating to its iron-chelating activity. AB - OBJECTIVE: To investigate the in vitro antimalarial effect of daphentin relating to its iron-chelating activity. METHODS: Schizontocidal activity of daphnetin and desferrioxamine B was tested through an in vitro assay based on the routine in vitro cultivation of Plasmodium faciparum FCC1 strain. The iron-chelating ability of each was measured by the fluorescent probe calcein. RESULTS: Daphnetin exhibited a modest iron-chelating ability compared with the powerful iron chelator desferrioxamine B. In vitro test at the range of 0-12 mumol/L daphnetin showed a dose-dependent schizontocidal activity which could be inhibited if mixed with Fe2+ in a ratio of 2:1. CONCLUSION: The dose-dependent antimalarial activity of daphnetin is related to its iron-chelating activity. PMID- 12567566 TI - [Sequence analysis of rDNA-LSU gene of Orientobilharzia turkestanicum from mainland of China]. AB - OBJECTIVE: To classify the taxonomic status of O. cheni in relation to O. turkestanicum var. tuberculata from the mainland of China by comparing their nucleotide sequences of nuclear ribosomal partial large subunit gene (LSU). METHODS: The genomic DNA of adult worms were extracted by the GNT-K method. The target gene was amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid PCR-blunt (Invitrogen). Recombinant plasmids were amplified in E. coli, extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of O. turkestanicum was retrieved from GenBank and aligned with our data in BioEdit. RESULTS: The nucleotide sequences of LSU between O. turkestanicum var. tuberculata and O. cheni was 100% identical, and 99.99% identical between O. turkestanicum var. tuberculata and O. turkestanicum. CONCLUSION: This study demonstrated high similarity in LSU nucleotide sequences, and the results do not support O. cheni as an independent species. O. cheni may be a synonym of O. turkestanicum var. tuberculata, and O. turkestanicum var. tuberculata is probably also a synonym of O. turkestanicum. PMID- 12567567 TI - [Study on low density lipoprotein receptor of adult Schistosoma japonicum]. AB - OBJECTIVE: To provide a theoretical basis for the study of vaccines against Schistosoma japonicum, the receptor for human LDL in the tegument of adult Schistosoma japonicum was investigated. METHODS: Proteins existed in adult Schistosoma japonicum membrane were extracted by Triton X-100 and purified through reverse-phase high performance liquid chromatography (HPLC), and the main protein peaks were then collected separately. 125I-LDL of human plasma as the ligand, through the radioautography and radioligand binding assay, the protein which can bind human serum 125I-LDL specifically was identified. The molecular weight and IEF were detected by SDS-PAGE. RESULTS: According to the radioautography and radioligand binding assay, a protein with retention time of 10.5 min was proved to be able to bind human serum 125I-LDL specifically. SDS PAGE revealed that the molecular weight of the purified protein is 60-65 kDa, and its IEF is 6.7. CONCLUSION: LDL binding protein may exist on the surface of both male and female adult Schistosoma japonicum with the function of obtaining cholesterol from host circulating system. PMID- 12567569 TI - [Surveillance of schistosomiasis in flatlands of mountain region]. PMID- 12567568 TI - Assessment of therapeutic efficacy of chloroquine against falciparum malaria in an outbreak area in Xishuangbanna, Yunnan. AB - OBJECTIVE: To investigate the failure of treatment with chloroquine in Yunnan in order to help formulate adequate antimalarial drug policy. METHODS: A World Health Organization 28-day in vivo test on therapeutic response for uncomplicated falciparum malaria in area with low or moderate transmission was adopted. Patients of age > or = 6 months old were admitted without limitation in density of parasitaemia and body temperature. Clinical and parasitological observation was conducted for patients on day 0, 1, 2, 3, 4, 7, 14, 21, 28 and 35. RESULTS: Of 62 patients identified as malaria cases infected by Plasmodium falciparum only, Plasmodium vivax only or by both species, 52 cases infected by Plasmodium falciparum only were included in the study. The overall treatment failure rate was 40.7%, with early treatment failure (ETF) rate of 1.8% and late treatment failure rate (LTF) of 38.9%. CONCLUSION: The treatment failure rate was much higher than the rate of 25% recommended by WHO. It is suggested that use of single chloroquine should be stopped in the treatment of falciparum malaria cases in such area. No relationship was found between the failure rate and the density of malaria parasites. PMID- 12567570 TI - [Study on number and mature rate of eggs in gravid proglottids of Taenia solium]. AB - OBJECTIVE: To investigate the number and mature rate of eggs in gravid proglottids of Taenia solium. METHODS: Ten worms of Taenia solium, expelled from patients, were detected. Eggs were collected from the last 10 gravid proglottids of each worm. RESULTS & CONCLUSION: The egg number in each mature proglottids varied from 3,900 to 126,520, and the mean number was 28,332. The mature rate of eggs was from 7.00% to 36.00% with an average of 29.12%, which was lower than that in proglottids naturally excreted with feces. With suitable temperature and humidity, the proglottids developed continually after excreted out of host body. Two to three days later, the mature rate of their eggs increased to 85%-90%. PMID- 12567571 TI - [Sensitivity of Oncomelania snail to niclosamide in China]. AB - OBJECTIVE: To understand the variation in response of Oncomelania hupensis to niclosamide. METHODS: Snails were collected from 37 sampling areas distributed in 10 provinces (municipalities) using random environmental sampling methods in accordance with the different types and categories of snail habitats. In laboratory the snails were immersed in solutions of niclosamide for 24 and 48 hours at 25 degrees C. RESULTS: 1.0 mg/L niclosamide showed 100% killing effect on snails in 24 hours. The LC50 concentrations for snails immersed for 24 hours ranged from 0.0320 to 0.1689 mg/L with a mean value of 0.0920 mg/L. 0.5 mg/L niclosamide showed 100% killing effect on snails in 48 hours. The LC50 values for snails immersed for 48 hours ranged between 0.0299 and 0.1114 mg/L with a mean of 0.0627 mg/L. There is a significant difference in snail sensitivity to niclosamide between sampling areas. CONCLUSION: The sensitivity to niclosamide varied in snails from different sampling fields, but the chemical in a concentration of 1.0 mg/L showed 100% effect of killing snails, which is consistent to the manual of schistosomiasis control. PMID- 12567572 TI - [A young child infected with (Taenia solium)]. PMID- 12567573 TI - [Prevalence survey on Cyclospora cayetanensis and Cryptosporidium ssp. in diarrhea cases in Yunnan Province]. AB - OBJECTIVE: To investigate the prevalence and distribution characteristics of Cyclospora cayetanensis and Cryptosporidium ssp. infection in diarrhea cases of Yunnan Province. METHODS: To collect fresh faeces from diarrhea cases in 7 counties/cities, examine the specimens by direct smear with iodine staining and modified acid-fast staining. RESULTS: The infection rate of C. cayetanensis and Cryptosporidium ssp. was 3.97% and 5.29%, respectively. The infection rate of the two pathogenic coccidians was as high as 10.64% and 8.51% in preschool children. C. cayetanensis was found in 3 counties and Cryptosporidium in 6 counties. CONCLUSION: Both C. cayetanensis and Cryptosporidium ssp. are prevalent in Yunnan Province with the latter distributed more widely, and the two pathogens are more prevalent in preschool children. PMID- 12567574 TI - [Evaluation of Gd-DTPA enhanced MR in the diagnosis of the degeneration stage of cerebral cysticercosis]. AB - OBJECTIVE: To detect the value of qualitative and orientating diagnosis in the degeneration stage of cerebral cysticercosis with Gd-DTPA enhanced MR. METHODS: Sixty-nine cases of cerebral cysticercosis were diagnosed by enhanced MR as degeneration stage, and confirmed by immunological examination and/or by surgery. MR plain scanning was conducted for the same cases. RESULTS: The plain scanning showed single or multiple lesions with long T1 and long T2 signals, and the enhanced scanning showed nodular or annular lesions. The diameter of the lesion after enhanced scan was not more than 22 mm with an average value of 8.1 mm. Some cases showed single lesion on plain scanning but showed multiple lesions after enhanced scan. CONCLUSION: The enhanced MR shows more typical features of the degeneration stage cerebral cysticercosis. It can define the number, position and range of the lesions, and can improve the accuracy of differential diagnosis, and therefore be of importance in formulating treatment scheme and prognosis. PMID- 12567575 TI - [A misdiagnosed case of human fascioliasis hepatica]. PMID- 12567576 TI - [Advance on classification of cyst-forming Coccidia and it's systematic development]. PMID- 12567577 TI - [Peroxiredoxin and parasite]. PMID- 12567578 TI - [In vitro response of Plasmodium falciparum FCC1/HN to artesunate, pyronaridine and other antimalarials]. PMID- 12567579 TI - [Construction of Schistosoma japonicum DNA vaccine candidate]. PMID- 12567580 TI - [An analysis for immuno-misdiagnosis of human hydatid disease]. PMID- 12567581 TI - [Recombinant construction of the gene encoding signaling protein of Schistosoma japonicum]. PMID- 12567582 TI - [A pilot investigation on caring of chronic filariasis cases]. PMID- 12567583 TI - [Two sorts of botanic granules misdiagnosed as parasite cases]. PMID- 12567584 TI - [Observation on the inducers for malaria gamete development in vitro]. PMID- 12567585 TI - [Malaria survey in Nangiao Township of Hainan Province]. PMID- 12567586 TI - Therapeutic options in the management of glaucoma simplex in the third millennium. PMID- 12567587 TI - Phacoemulsification in patients with secondary glaucoma caused by uveitis. AB - PURPOSES: To evaluate the effect of phacoemulsification in the patients with uveitis, secondary glaucoma and complicated cataract. METHODS: Phacoemulsification and implantation of a foldable intraoucular len was performed in 12 patients(13 eyes) with uveitis, secondary glaucoma and complicated cataract. The complications, intraocular pressures (IOP), and visual acuity were observed postoperatively. RESULTS: No severe complication was found in the patients postoperatively or within the operation procedure. The visual acuity was improved after the operation (P < 0.05). The intraocular pressures and/or the number of antiglaucoma medications reduced in 3 months of the follow-up time. CONCLUSION: Phacoemulsification is the best way to treat the patients with secondary glaucoma and complicated cataract caused by uveitis. PMID- 12567588 TI - Histopathological study of trabeculum after excimer laser trabeculectomy ab interno. AB - PURPOSE: To study the clinical manifestations and histopathologic changes of trabeculum after excimer laser trabeculectomy ab interno (ELT), and to investigate the mechanisms of ELT in reducing intraocular pressure. METHODS: ELT was performed on ten rabbit eyes and postoperative responses were documented. Corneoscleral tissue samples were harvested consecutively each week until the 5th postoperative week and these samples were examined under light and electrical microscopy. RESULTS: Mild stimulation signs were present postoperatively in nine of ten eyes, but no serious complications were experienced. Obvious inflammation was observed in one rabbit eye as a result of iris damage during the surgical manipulations. Local fractures on the trabecular meshwork and openings into Schlemm's canal were detected in all tissue samples under light microscope. Mitochondria were found to be turgescent and dilated like vacuoles and endoplasmic reticula were found to be dilated under electrical microscope in the early postoperative period. Later, all trabecular cells returned normal and no fibroblast cells were ever detected. CONCLUSIONS: Permanent openings through trabecular meshwork into the inner wall of Schlemm's canal can be created with ELT. The outflow resistance of aqueous humor can be reduced with these openings and intraocular pressure can be controlled thereafter. PMID- 12567589 TI - Experimental study on the effect of perfluorodecalin on rabbit corneas. AB - PURPOSE: To study the effects of perfluorodecalin on the cornea of the rabbit eyes. METHODS: Perfluorodecalin (0.05 ml/each) was injected into the anterior chambers of eighteen rabbit eyes. Corneal morphology and endothelial cells were monitored clinically by slit-lamp biomicroscope and specular microscope for 26 weeks. Animals were sacrificed in 1st, 2nd, 4th, 10th, 16th, 22nd, and 26th week after injection, respectively, and the corneas were examined under the light microscope. RESULTS: Perfluorodecalin droplets looking like "fish eggs" were found at about 1/4-1/2 of the corneal height in the inferior anterior chamber. Corneal opacification on the area contacted with perfluorodecalin was observed in five eyes five weeks after injection, and all in the 22nd week. Mutton fat KPs in one eye were seen in the 6th week firstly, and in all eyes in the 7th week. Corneal pannus formation in one eye was present in the 4th week, two eyes in the 5th week and three eyes in the 6th week. Retrocorneal fibrous membrane in one eye was detected at the 6th week and 3 eyes at the 7th week respectively. After injection of perfluorodecalin, endothelial cell density was significantly decreased (2,789 +/- 192 vs. 2,341 +/- 658, P < 0.01) and corneal thickness was increased. CONCLUSIONS: Perfluorodecalin injected into anterior chamber can lead to corneal damage and inflammatory reaction. PMID- 12567590 TI - Experiment study of effect of perfluorohexyloctane on corneal endothelial cells. AB - PURPOSE: To investigate the effect of Perfluorohexyloctane (F6H8) on corneal endothelial cells(CEC) of rabbit eyes. METHODS: Fifteen New Zealand white rabbits were divided into two groups: experimental group(F6H8) and control group(BSS). All rabbits underwent anterior chamber injection of 0.15 ml F6H8 or BSS. Slit lamp biomicroscopy and corneal endothelium photography were performed pre operatively and postoperatively. Histopathological examination and Transmission electron microscopy(TEM) were done after the rabbits were sacrificed. RESULTS: All the corneas were clear. Since 4 weeks after operation, the endothelial cells were markedly irregular in size and shape and the number of endothelial cells was markedly decreased. Multilayered retrocorneal membranes(RCM) grew gradually 2 weeks after surgery. Vacuolar degeneration was seen in some endothelial cells. Nuclear degeneration and edema of plasma were seen in TEM. CONCLUSION: Corneal endothelial cell degenerated after contacting with F6H8 for 2-4 weeks. As a silicone solvent, it should be removed completely after injection. We don't recommend it to be used as a new intraocular temponade. PMID- 12567591 TI - Dexamethasone modulation on cultured human retinal pigment epithelial cell. AB - PURPOSE: Dexamethasone(DEX) was tested for its ability to modulate human retinal pigment epithelium(hRPE) cell proliferation in cell culture. METHODS: DEX in different concentrations was added to cultured hRPE cells. The effects were measured with MTT method, 3H-thymidine(3H-TdR) incorporation and flow cytometry. RESULTS: DEX increased survival rate and DNA synthesis from 32 mg/L to 320 mg/L under hRPE culture conditions, but paradoxically reduced them at 1,000 mg/L and 3,200 mg/L in dose and time dependent fashion by both MTT assay and 3H-TdR incorporation. The cell numbers in S phase and G2/M phase increased 28.32% at DEX concentration 320 mg/L, in contrast, reduced 41.84% at 1,000 mg/L. CONCLUSION: DEX increased proliferation from 32 mg/L to 320 mg/L, and inhibited proliferation at concentrations greater than 320 mg/L. The inhibiting effect of DEX may happen in s phase and G2/M phase. PMID- 12567592 TI - Clinical analysis of Leber's hereditary optic neuropathy harboring mtDNA mutation at nt11778. AB - PURPOSE: To improve our diagnostic technique through the analysis of clinical features of Leber's hereditary optic neuropathy (LHON) harboring mtDNA point mutation at nt11778. METHODS: Detection of nt11778 mutation was performed on 38 patients clinically diagnosed as LHON in our ophthalmic center from year 1998 to 2000. Circumstances of onset and family history were obtained and ophthalmoscopy, fundus fluorescein angiography, visual field and visual evoked potential were performed on all 38 patients. RESULT: 30 In 38 patients (78.95%) harbor nt11778 mutation, including 28 male (93.33%) and 2 female (6.67%). The ratio of affected male to female is 14:1. Patients harboring nt11778 mutation display typical clinical manifestations. CONCLUSION: Identification of one of the three LHON specifically associated mtDNA mutations is essential to confirm the diagnosis. PMID- 12567593 TI - Management of post-traumatic aniridia with retinal detachment. AB - PURPOSE: To reconstruct the anatomic and functional impairment in patients with post-traumatic aniridia, aphakia, and retinal detachment. METHODS: Four patients with unilateral aniridia and aphakia as well as retinal detachment as results of severe eye injuries underwent scleral buckling, vitrectomy, membrane peeling, endolaser photocoagulation, silicone oil or gas temponade, combined with iris diaphragm-IOL implantation. RESULTS: All four patients achieved successfully anatomic and functional reconstruction after surgery. During five to 22 months postoperative follow-up, all retinas remained attached. The final visual acuity increased from finger counting to 0.1-0.3. CONCLUSIONS: The combination of vitreoretinal surgery and iris diaphrgm-IOL implantation is an effective method for post-traumatic aniridia, aphakia and traumatic retinal detachment. It could ameliorate photophobia and improve the biocular vision. Furthermore, artificial iris diaphragm implantation could prevent silicone oil-endothelia contact and salvage silicone keratopathy. PMID- 12567594 TI - Clinical observation on preservation of lens anterior capsule in reduction of complications associated with silicone oil. AB - PURPOSE: To observe the effect of preservation of anterior capsule on the incidence of complications associated with silicone oil. METHODS: 82 patients (82 eyes) accepted trans pars plana vitrectomy (TPPV) combined with lensectomy of whom, 30 eyes with preservation of anterior capsule (PAC) and 52 eyes with no preservation of anterior capsule (NPAC). Analysis of the incidence of complications associated with use of oil. RESULTS: The incidence rate was 50.0% in NPAC group, and 23.3% in PAC group (P < 0.025). There were secondary glaucoma (21.1%), band keratopathy (13.5%) and corneal decompensation (9.6%) in NPAC group, while there was none of them in PAC group. CONCLUSIONS: Preservation of anterior capsule is an effective method to reduce the complications associated with silicone oil. PMID- 12567595 TI - Effect of 0.2% brimonidine on retinal blood flow. AB - PURPOSE: To analyze the effect of 0.2% brimonidine eye drops on retinal blood flow of patients with glaucoma. METHODS: Using self-control method and Heidelberg Retina Flowmeter (HRF), we examined the volume, flow and velocity of the superior nasal and temporal, the inferior nasal and temporal artery of retina at baseline and 2 hours after single instillation of 0.2% brimonidine. RESULTS: There were no significant changes in volume, flow and velocity of four vessels before and after the administration of 0.2% brimonidine. CONCLUSIONS: There are no significant ocular haemodynamic benefits associated with Brimonidine therapy to the glaucoma patients. PMID- 12567596 TI - Change of visual acuity and related factors in idiopathic optic neuritis. AB - PURPOSE: To study the change of visual acuity and related factors in idiopathic optic neuritis. METHODS: Retrospectively studying 136 cases (210 eyes) of idiopathic optic neuritis hospitalized in our department during last 5 years. The data was analyzed with the methods of multivariate stepwise regression analysis by SPSS statistic package. RESULTS: The final visual acuity was very strongly related positively to the initial visual acuity at the time point of starting treatment and negatively to the time span beginning the treatment to the time showing improvement, age of patient and the duration from onset of the disorder to the time point of treatment beginning. CONCLUSIONS: Promptly right treatment can significantly change the natural course and the final vision of optic neuritis. Severe visual loss, old age, longer time without right therapy and slow improvement after the treatment may indicate an unfavorable final visual recovery. The different dosages of steroid applied in this study was not significant in view of the final visual acuity. PMID- 12567597 TI - The electroretinogram in patients with retinal vascular occlusion. AB - PURPOSE: To measure and analyze the electroretinogram (ERG) in patients with retinal vascular occlusion. METHOD: Fifty-eight cases (59 eyes) of retinal vascular occlusion and the fellow eyes of 47 cases were tested with Ganzfeld ERG. The scotopic and photopic ERG, and oscillatory potentials were tested according to the ERG standard of ISCEV. RESULTS: The abnormal rates of OPs and b wave were higher in retinal vascular occlusion. The abnormal ERG appeared mostly in CRVO among four types of retinal vascular occlusion. The comparisons of some amplitudes and latencies between the fellow eyes and the affected eyes showed statistically significant difference. CONCLUSION: The oscillatory potentials and b waves have important roles in evaluating the retinal function of patients with retinal vascular occlusions. There are some clinical significance for comparing ERG between the affected eye and the fellow eye. PMID- 12567598 TI - Foldable lens explantation and exchange: the reason and solution. AB - OBJECTIVE: To report the explanation and exchange of Hydrophilic Acrylic foldable intraocular lens (IOL) on 14 patients who had visual disturbances caused by the change of transparence on optic. METHODS: Sixteen Hydrophilic Acrylic foldable intraocular lenses from 14 patients who presented with decreased visual acuity from 6 months to 1 year after normal phacoemulsification and IOL implantation associated with extensive transparent change on optic of the lens. The lenses were explanted with the bisection technique. All the eyes were reinserted with Acrysof foldable lenses. RESULTS: Sixteen lenses were removed successfully and exchanged with the new lens in the capsule. The posterior capsular rupture and vitreous loss were found in the first two cases. One of them had the zonulysis due to the radial tear of the anterior capsule during the enlargement of the capsular opening. The anterior vitrectomy was performed with IOL fixed on the ciliary sulcus. The visual acuity of all the patients improved obviously without posterior complication. CONCLUSION: Foldable lens explanation with the bisection technique and exchange had a successful outcome with improvement of ocular condition. PMID- 12567599 TI - The preliminary clinical observation of array multifocal intraocular lens implantation. AB - PURPOSE: To evaluate the clinical effects of implantation of Array multifocal intraocular lenses. METHODS: Thirty-one cases (37 eyes) of cataract patients, including 15 males (19 eyes) and 16 females (18 eyes), were involved in this study. All patients underwent standard phacoemulsification with Array multifocal intraocular lens implantation. The complications during operation, postoperative distant visual acuity, near visual acuity, corneal curvature and visual symptoms were observed. RESULTS: The mean value of best postoperative visual acuity was recorded as follows: uncorrected distant visual acuity was 0.8, the best corrected distant visual acuity was 0.9, uncorrected near visual acuity was 0.5, near visual acuity with distant-corrected was 0.6, the best-corrected near visual acuity was 0.9. The astigmatism of cornea was less than 1.5 D pre-operatively and post-operatively. One patient complained of glare. CONCLUSION: Array multifocal intraocular lens can provide good distant and near visual acuity. With observation of more cases and follow-up of longer time, we can draw a further conclusion. PMID- 12567600 TI - Comparison of trans-scleral fixation of PMMA and foldable intraocular lens in children. AB - PURPOSE: To observe the difference of the effects of PMMA and foldable intraocular lenses (IOLs) trans-sclerally fixed in pediatric eyes. METHODS: Thirty-two children (43 eyes) who had undergone trans-scleral fixation of IOL were retrospected, of whom 5 children were implanted PMMA IOL in both eyes, 6 children were implanted PMMA IOL in one eye and foldable IOL in the other eye, 12 children were implanted foldable IOL in one eye and 9 children were implanted PMMA IOL in one eye. Mean age was 5.3 years (range 2.5-12 years). Twelve children had traumatic cataract and the others congenital cataract before lens extraction. RESULTS: Foldable group (18 eyes): Mean follow-up was 12.1 months. Visual acuity (VA): compared with the best corrected VA before IOL fixation, postoperative best corrected VA improved in 16 eyes, remained unchanged in 2 eyes. In 14 eyes, one or two stitches were needed to seal the incision. COMPLICATIONS: Severe anterior chamber reaction was seen in 3 eyes. Intraocular bleeding was found in 3 eyes. IOL decentration was detected in 1 eye. Iris capture of IOL was seen in one eye. PMMA group (25 eyes): Mean follow-up was 20.3 months. Visual acuity (VA): compared with the best corrected VA before IOL fixation, postoperative best corrected VA improved in 19 eyes, remained unchanged in 5 eyes and got worse in one eye. In 24 eyes, one to three stitches were needed to seal the incision. COMPLICATIONS: Severe anterior chamber reaction was seen in 5 eyes. Intraocular bleeding was found in 4 eyes. IOL decentration was seen in one eye. Iris capture of IOL was seen in 3 eyes. Intraocular pressure elevated in one eye. CONCLUSION: Our study shows that trans-scleral fixation of IOL is a safe procedure in pediatric eyes. Foldable IOL showed similar effect compared with PMMA IOL in pediatric trans-scleral fixation. PMID- 12567601 TI - [Sequence analysis of SSU rDNA variable regions of Leishmania isolates from hilly foci and plain foci of China]. AB - OBJECTIVE: To analyze the sequence difference of the SSU rDNA variable regions of Leishmania isolates from hilly foci and plain foci of China. METHODS: Specific SSU rDNA fragments from nuclear DNA of five Leishmania species and isolates were amplified by PCR. The amplified DNA fragments were cloned into pGEMR-T Easy vector. The specific fragments were sequenced by the automated DNA sequencer. RESULTS: Sequence analysis showed that the amplified DNA fragments of five Leishmania species and isolates were all 392 bp in length, point mutations were located in the two unique sequence (UQ-I and UQ-II); L. d. SC10 and L. d. GS7 had two same point mutations in UQ-II, only L. d. GS7 had one in UQ-I; no insertion/deletion. CONCLUSION: Sequence difference of the SSU rDNA variable region existed between Leishmania isolates from hilly foci and plain foci; The sequences of the SSU rDNA variable regions of L. d. SD2 isolate and L. infantum were identical. PMID- 12567602 TI - [On molecular identification and taxonomic status of Anopheles lesteri and Anopheles anthropophagus in China (Diptera: Culicidae)]. AB - OBJECTIVE: To clarify the taxonomic status of Anopheles lesteri and An. anthropophagus in China. METHODS: Using molecular identification (PCR assay and rDNA-ITS2 sequencing) to examine the field anopheline mosquito specimens from Liaoning and Shandong. According to the ITS2 sequences, molecular phylogenetic tree was made. RESULTS: According to the molecular identification, An. lesteri and An. anthropophagus were distributed both in Liaoning Province and Shandong Province. The length and GC content of rDNA-ITS2 sequence were 451 bp, 46.2% in An. lesteri (n = 6), and 448 bp, 46.0% in An. anthropophagus (n = 10), respectively. The ITS2 sequences from presentation sites were same in An. lesteri, while the intraspecies difference in An. anthropophagus was 0.88%. The specific difference between An. lesteri and An. anthropophagus was 25.7%. By analyzing molecular phylogenetic tree, the relationship between An. lesteri and An. sinensis, An. anthropophagus and An. liangshanensis was found to be closer. CONCLUSION: According to the molecular identification, it was defined that An. lesteri and An. anthropophagus were sympatric independent species in China. PMID- 12567603 TI - [Immune response in mice induced by C terminal encoding gene of Plasmodium falciparum histidine rich protein. 2]. AB - OBJECTIVE: To explore the humoral and cellular immune responses in mice to eukaryotic expression recombinant plasmid encoding histidine rich protein 2 (HRP II) of Plasmodium falciparum. METHODS: The start and stop codes were introduced into HRP-II gene fragment, the reading frame and the position of start and stop codes in HRP-II were identified by sequencing. HRP-II fragment containing the start and stop codes was cloned into pcDNA3.1 (-) to form pcDNA3.1 (-)/HRP-II. The BALB/c mice were immunized i.m. with the plasmids for 3 times in 3 weeks intervals. Two weeks after the last immunization, the sera and splenocytes were collected to investigate anti-HRP-II antibodies by ELISA and the splenocytes proliferation response to HRP-II. RESULTS: Sequence data show that the reading frame and the position of start and stop codes are correct. Restriction enzyme digestion indicated that the HRP-II gene fragment containing start and stop codes was successfully cloned into pcDNA3.1 (-). Mice raised significant anti-HRP-II antibodies after pcDNA3.1 (-)/HRP-II immunization, and the splenocytes proliferated prominently when stimulated with HRP-II protein. CONCLUSION: Eukaryotic expression recombinant plasmid encoding HRP-II gene can induce significantly humoral and cellular immune response in mice. HRP-II gene may be a good candidate for P. falciparum blood-stage multiple DNA vaccine. PMID- 12567604 TI - [Clinical analysis of 70 cases of cysticercosis]. PMID- 12567605 TI - [Preparation and characterization of McAbs against lactate dehydrogenase of Plasmodium falciparum]. AB - OBJECTIVE: To prepare and characterize the monoclonal antibodies(McAbs) against lactate dehydrogenase of the Plasmodium falciparum(LDHp). METHODS: BALB/c mice were immunized with purified recombinant LDHp and McAbs against LDHp were prepared according to the protocol of hybridoma technique. The McAbs were characterized by ELISA and Western blot analysis. RESULTS: Two McAbs against LDHp antigen were obtained. Both McAbs were IgG2b. The titer of two McAbs(2A5, 1H10) in the ascites was 1:25,600 and 1:12,800, and in supernatant were 1:512,1:256 respectively. The result of ELISA indicated that two McAbs reacted only with P. falciparum, and did not react with normal human red blood cells, P. vivax, Toxoplasma gondii, Schistosoma japonicum. It is recognized 33 kDa protein which was defined as LDHp by Western blot analysis. CONCLUSION: Two hybridoma cell lines secreting high titer of McAbs against LDHp with high specificity were established. PMID- 12567606 TI - [A case of sparganosis mansoni]. PMID- 12567607 TI - [Effect of artemether on phosphoglucomutase, aldolase, phosphoglycerate mutase and enolase of Schistosoma japonicum harbored in mice]. AB - OBJECTIVE: To study the effect of artemether (Art) on phosphoglucomutase (GPM), aldolase (ALD), phosphoglycerate mutase (PGM) and enolase (ENO) of Schistosoma japonicum harbored in mice. METHODS: Mice infected with S. japonicum cercariae for 4-5 wk were treated ig with Art 100 mg/kg or 300 mg/kg and killed 24 h or 48 h after treatment for collection of worms. The activities of GPM, ALD, PGM and ENO in female and male worms were measured by the formation of NADPH or consumption of NADH. RESULTS: After the worms were exposed in vivo to Art 100 mg/kg for 24 h, the GPM, ALD, PGM and ENO activities in female worms were significantly decreased by 15%, 19%, 50% and 46%, respectively, while in male worms only the PGM and ENO activities were markedly decreased by 22% and 32%, respectively. Following exposure of the worms to Art 100 mg/kg for 48 h, the GPM and ALD activities in male worms were also significantly reduced by 21% and 18%, respectively, while the activities of GPM, ALD, PGM and ENO in female worms and those of PGM and ENO in male worms declined progressively with time. After the worms were exposed in vivo to Art 300 mg/kg for 24-48 h, all the activities of the above-mentioned enzymes in female and male worms declined significantly in a time-related pattern. CONCLUSION: Art showed an apparently inhibitory effect on GPM, ALD, PGM and ENO in female schistosomes. PMID- 12567608 TI - [Cloning and sequence analysis of RESA gene fragment of Plasmodium falciparum isolate FCC1/HN]. AB - OBJECTIVE: To determine the nucleotide sequence of the 3'-termal of the RESA gene Plasmodium falciparum isolate FCC1/HN, and find out the differences of the sequences of RESA gene among isolate FCC1/HN, FC27, NF7 and Palo Alto. METHODS: 3'-terminal fragment of RESA gene of P. falciparum isolate FCC1/HN was amplified by PCR method, then was cloned into pMD18-T vector. The recombinant was screened and identified by BamHI + XhoI and PCR technique. The nucleotide sequence of the 3'-terminal of the RESA gene was determined by the dideoxy chain termination method. DNASTAR and BLAST software were used to compare and analyze the RESA gene sequences among the different isolates. RESULTS: The 3'-termal fragment of the RESA gene with about 846 bp was specifically amplified by PCR, the recombinant pMD18-T-RESA was successfully constructed. Different degrees of diversity of the RESA gene sequences were found among P. falciparum isolates FCC1/HN, FC27, NF7 and Palo Alto. CONCLUSION: There were differences in the sequences of RESA gene among the P. falciparum isolate FCC1/HN and three other isolates (FC27, NF7 and Palto alto). PMID- 12567609 TI - [Amplification, cloning and expression of a gene encoding hexose transporter of Plasmodium falciparum]. AB - OBJECTIVE: To amplify, clone and express of a gene encoding hexose transporter of Plasmodium falcipuram (PfHT1) from Southern China isolate FCC1/HN for studing the immune of recombinant which protective from malaria parasite infection. METHODS: Cultivation of P. falciparum isolate FCC1/HN in vitro; extraction of genomic DNA from FCC1/HN using the alkali specific cleavage method; PCR amplification of PfHT1 and cloning into eukaryotic expression vector, pEGFPN3. The recombinant as introduced into mammalian cells, HEPG2 by using liposome-mediated transfection. RESULTS: The gene encoding PfHT1 was specifically amplified from the genomic DNA of P. falciparum isolate FCC1/HN. The size of amplified fragment was 1,516 base pair. The eukaryotic expression recombinant, pN3-HT1, was constructed and expressed steadily in the hepatocarcinoma cell lines, HEPG2. CONCLUSION: The gene encoding PfHT1 was successfully amplified and cloned. The pN3-HT1/HEPG2 cell line was built for expressing fusion protein of GFP-HT1. PMID- 12567610 TI - [Correlation analysis of the malaria control measures and the number of malaria cases in Neijiang City]. PMID- 12567611 TI - [Analysis of trace elements in liver, spleen and brain of rats infected with Toxoplasma gondii]. AB - OBJECTIVE: To study the level of five trace elements(Fe2+, Cu2+, Zn2+, Ca2+, Mg2+) in the liver, spleen and brain of the rats infected with Toxoplasma gondii. METHODS: 20 rats were randomly and equally divided into two groups: normal group and experimental group. On the 64th day after injection of 1.5 x 10(6) T. gondii/2 ml, all of the rats were killed and the atomic absorption method were used. RESULTS: Compared with the normal group, the level of Cu2+ in liver in experimental group was significantly reduced (P < 0.01), and the amount of Fe2+ of sick rats were higher than that of the normal (P < 0.01). No differences in the content of Zn2+, Ca2+, Mg2+ between the experimental and control group were determined; the level of Fe2+, Cu2+ in spleen in experimental group was significantly altered (P < 0.01). The amount of Mg2+ of sick rats were higher than that of the normal (P < 0.01), and no difference on the content of Zn2+, Ca2+ between the experimental and control group were estimated; the level of Fe2+, Cu2+, Mg2+ in brain in experimental group was significantly reduced (P < 0.01). The amount of Ca2+ of sick rats were higher than that of the normal (P < 0.02). No difference on the content of Zn2+ between the experimental and control group were estimated. CONCLUSION: T. gondii infection might cause changes in trace elements in the liver, spleen and brain of rats. PMID- 12567612 TI - [Longitudinal observation on the control of intestinal helminthiasis]. AB - OBJECTIVE: To search suitable measure for rapid control intestinal helminthiasis and long-term strengthen efficacy. METHODS: The treatment was taken in egg positive population of intestinal helminthiasis in 1986-1988. The treatment was carried out only in the selected population in 1989-1992. No measure was taken in 1993-2000. RESULTS: (1) The prevalence rate of hookworm, Ascaris and Trichuris decreased to 3.2%, 37.3% and 3.5% respectively after administration of albendazole twice a year for 3 years. (2) The prevalence rate of hookworm continued to decrease to 0.5% after treatment on selected population. (3) The prevalence rate and the intensity of hookworm has been less than 1% and 10/LPG for 8 years. No hookworm larvae had been isolated from the soil. CONCLUSION: The hookworm transmission was effectively controlled in the study site. PMID- 12567613 TI - [Observation on the effect of certain Chinese herbs against the intestinal parasites and mites in mice]. PMID- 12567614 TI - [Surveillance of filariasis in some vulnerable areas in Guangxi after filariasis elimination]. AB - OBJECTIVE: To explore the measures for continuing surveillance of filariasis. METHODS: Selecting some vulnerable spots for focal surveillance, double-slide biood sampling for microfilaria examination, dissecting vectors for detecting the mosquitoes infected with filarial larva, using IFAT for detecting antifilarial antibody. RESULTS: 27,938 persons were examined for microfilaria and 4,454 mosquitoes were dissected for filaria larva, all were negative. 3,606 serum samples were examined for antifilarial antibody average positive rate was 1.35%(0.39-4.97%). CONCLUSION: The results of surveillance showed that the achievement of filariasis control in Guangxi after filariasis elimination is consolidated. PMID- 12567615 TI - [Clinical analysis of 36 cases with amebic liver abscess]. AB - OBJECTIVE: To investigate the clinical features of amebic liver abscess, the causes of misdignosis and the effect of medical and surgical therapy on patient's prognosis. METHODS: The clinical features of 36 patients with amebic liver abscess admitted from 1982 to 1997 in our hospital were retrospectively reviewed. RESULTS: The major clinical manifestations were: abdominal pain (86.1%), fever (86.1%), hepatomegaly with tenderness (83.3%) and right intercostal tenderness (58.3%). Leukocytosis was observed in 61.1%, and increased of ESR in 88.5% (23/28). Serologies against Entamoeba histolytica were noted in 92.6%. Ultrasonography showed single lesions in 75% and right-lobe involvement in 75%. All patients were treated with metronidazole and 27 patients received treatment with needle aspiration or draining at the same time. After treatment, 10 patients were cured, 25 patients were improved significantly and effective rate was 97.2%. One patient died of hepatic failure. CONCLUSION: Medical therapy alone was excellent for small abscesses, while percutaneous needle aspiration or draining was a successful approach in patients with large abscesses. PMID- 12567616 TI - [Immunoregulation of CD4 + T cells in intraerythrocytic malaria parasite infection]. PMID- 12567617 TI - [On inducible variations in genetics of malaria parasites from the view of emergence of chloroquine]. PMID- 12567618 TI - [Investigation of intestinal parasitic infections in the countryside of Dafeng City]. PMID- 12567619 TI - [Malaria Internet resources helps the molecular biological research on malaria parasites]. PMID- 12567620 TI - [Observation on the effect of irrigation ditch reform and chemotherapy of humans and animals in the control of schistosomiasis]. PMID- 12567621 TI - [Clinical and CT analysis of 1062 cases with cerebral cysticercosis]. PMID- 12567622 TI - [Control measures and systematic surveillance of filariasis in Zhoushan City]. PMID- 12567623 TI - [Analysis of the malaria surveillance in Fujian Province in the past 20 years]. PMID- 12567624 TI - [Application of capture-recapture method to adjust the number of malaria cases from reporting data]. PMID- 12567625 TI - [Studies on the control strategy of acute schistosomiasis in Yunnan mountainous regions]. PMID- 12567626 TI - [Investigation of the distribution of Anthropophagus lesteri in Chongqing municipality]. PMID- 12567627 TI - [Cloning and sequence analysis of the light chain variable region gene of monoclonal anti-idiotypic antibody NP30 of Schistosoma japonicum]. AB - OBJECTIVE: To amplify and sequence the light chain of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum. METHODS: By comparing the conserved regions at each end of the nucleotide sequences of murine germ-line genes encoding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of VL gene. The hybridoma cells secreting anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger's method. The VL gene was compared with GenBank and published mouse VL genes. RESULTS: The full-length of VL gene was 318 bp. The VL gene was a member of mouse Ig kappa light chain subgroup IV and generated from rearrangement of germ line V and J kappa 4 genes. The VL gene sequence has been registered by GenBank(accession No. AF206720). CONCLUSION: The obtained VL gene was a potentially functional gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum. PMID- 12567628 TI - [A case of ocular Thelazia callipaeda infection]. PMID- 12567629 TI - [Cloning and sequence analysis of SSU rRNA gene of cutaneous leishmaniasis pathogen from Xinjiang of China]. AB - OBJECTIVE: By sequencing of SSU rRNA gene cloning from Xinjiang cutaneous leishmaniasis pathogen (XJCLP) to provide evidence for identification of the pathogen. METHODS: By PCR assay with primers R222 and R333, the specific fragment had been produced from SSU rRNA gene of XJCLP, L. infantum, L. tropica and cloned into pGEM-T Easy vector. The clones were sequenced by the Sanger dideoxy-mediated chain termination method, analysis of SSU rRNA gene sequences from XJCLP, L. tropica, L. infantum with DNASIS. RESULTS: Sequence analysis showed that the specific fragment of SSU rRNA gene from XJCLP, L. infantum, L. tropica, were all 394 bp in length. There were 391 bases identical and three point mutations between the sequences of XJCLP and L. tropica, the similarity being 99.2%; 390 bases identical and three point mutations and one insertion/deletion between the sequences of XJCLP and L. infantum, the similarity being 99.0%. One insertion/deletion between the sequences of L. tropica and L. infantum, the similarity being 99.7%. The primary and secondary structures of SSU rRNA gene from XJCLP differed from those of L. infantum and L. tropica. A retrieval from GenBank confirmed that these 394 bp sequence are new gene sequences. CONCLUSION: The primary and secondary structures of SSU rRNA gene from XJCLP, L. infantum, L. tropica were different. 394 bp sequence from SSU rRNA gene of XJCLP is a new gene sequence. PMID- 12567630 TI - [Cloning and identification of cytochrome P450 resistance related genes in the mosquito, Culex pipiens pallens]. AB - OBJECTIVE: To inquire into the relationship between cytochrome P450 and deltamethrin resistance. METHODS: 24 new cDNA sequences encoding cytochrome P450 were amplified respectively from deltamethrin-susceptible and -resistant strains of Culex pipiens pallens with a pair of degenerate primers according to the conservative amino acid sequences of CYP4 in insects by RT-PCR and the Direct Cloning Method, and then were identified by cDNA chip and reverse Northern. RESULTS: 112 positive clones were obtained, of which 24 were shown to be new sequences encoded for cytochrome P450. They have been lodged in GenBank and were appraised by the Nomenclature Committee of Cytochrome P450, belonging to the subfamily CYP4C, CYP4D, CYP4H and CYP4J in CYP4 family. The hybrid signal values of 6 P450 sequences (NYDS3, NYDS5, NYDR6, NYDR9, NYDR15 and NYDR17) were 3.1-9.7 times higher in the resistance probe than in the susceptible probe, and NYDR17 only reacted with the resistance probe. The result of reverse Northern in NYDR15 was similar to that of cDNA chip. CONCLUSION: CYP4 is related to deltamethrin resistance and the specific expression caused by point mutation of cytochrome P450 gene may exist in deltamethrin-resistant Cx. pipiens pallens. PMID- 12567631 TI - [A case of dermamyiasis infected with Hypoderma bovis larvae]. PMID- 12567632 TI - [Studies on apoptosis and its induction in Schistosoma japonicum]. AB - OBJECTIVE: To explore the apoptosis and its induction in S. japonicum. METHODS: The eggs and adult cells of S. japonicum were isolated. The adult cells were incubated with H2O2, dexamethasone and cyclosporin A, cyclosporin A vehicle solution, respectively in order to initiate the apoptosis. The eggs were treatd at 4 degrees C, 37 degrees C and 42 degrees C, respectively for induction of apoptosis. The occurrence of apoptosis was supervised by agarose gel electrophoresis and in situ end labelling. RESULTS: DNA ladder-band characteristic of apoptosis appeared in the cells of adult worms after incubated with H2O2, dexamethasone or cyclosporin A and the eggs treated at 42 degrees C, whereas no DNA ladder band was found in other control groups. In situ end labeling revealed that the positive fluorescence could be seen in the cells incubated with H2O2, dexamethasone or cyclosporin A and few nuclei in the control group and cyclosporin A vehicle solution group appeared positive fluorescence. CONCLUSION: Apoptosis in the cells of S. japonicum adults and eggs induced by chemical or physical agents was observed. PMID- 12567633 TI - [Studies on identification of circumsporozoite protein genotyping of Plasmodium vivax]. AB - OBJECTIVE: To develop a new method of genotyping circumsporozoite protein (CSP) gene for identification of field isolates of Plasmodium vivax. METHODS: Improved Chelex-100 ion-exchange method was used to extract DNA from blood filter paper samples, nested PCR and allele-specific PCR techniques, agarose gel electrophoresis analysis and dot/southern blotting-probe hybridization were employed for amplification, resolution and identification of the diagnostic fragments. RESULTS: Using the nest-allele-specific PCR assay reported here, small amounts of DNA extracted from a piece of blood filter paper sample were amplified which produced three different size ranges of diagnostic bands: 650-770 bp PV species-specific band, 170-230 bp diagnostic band for temperate zone family and 588 bp band for PV type-2. The sizes and patterns of the bands produced by the reference strains were consistent with those of designed target sequences. Of 59 examined isolates from 6 provinces of China, 42 temperate zone family strains, 15 tropical zone family strains and two PV type-2 strains were identified. CONCLUSION: 1, Three genotype strains of P. vivax mentioned above could be identified by this method with only two rounds of PCR and without probe hybridization. 2, The preliminary results showed that PV type-1 including temperate zone family strains and tropical zone family strains as well as PV type 2 strains are present in China. In addition, another CSP genotype with both sequence characteristics of temperate zone and tropical zone family might also be present in China. PMID- 12567634 TI - [Analysis of Pagumogonimus skrjabini antigen and its application in serodiagnosis]. AB - OBJECTIVE: To analyse the soluble antigens of different developmental stages of Pagumogonimus skrjabini and develop a specific and sensitive serodiagnostic method for pagumogonimiasis. METHODS: The soluble antigens of P. skrjabini of various stages were separated by SDS-PAGE. The specific antigen of the adult fluke was recognized immunologically by immunoblot assay. The protein bands between 10-30 kDa purified by SDS-PAGE and electrophoretic elution were used in dot-ELISA. RESULTS: Using dot-ELISA, the soluble antigens of adult were recognized by sera infected with P. skrjabini. More reactive bands appeared at 10 30 kDa, but major protein bands were at 22, 24 and 26 kDa. However, using sera from patients infected with other trematodes including schistosome and Clonorchis, cross-reaction bands appeared within 60 to 90 kDa. When compared with ELISA of crude adult antigens for detecting 28 suspected patients, there was no significant difference between the two methods. The sera of 38 patients with other diseases were also detected by the two tests. No cross-reaction occurred with the purified adult antigen dot-ELISA while 13.2% (5/38) of the sera cross reacted in ELISA of crude adult antigens. CONCLUSION: Dot-ELISA using 10-30 kDa antigen might be a specific and sensitive serodiagnostic method for diagnoing pagumogonimiasis. PMID- 12567635 TI - [Diagnosis of falciparum malaria using ICT]. PMID- 12567636 TI - [Studies on pharmacokinetics of chloroquine in mice infected with chloroquine resistant strain of Plasmodium berghei]. AB - OBJECTIVE: To compare the pharmacokinetic differences of chloroquine in normal mice and the mice infected with the N and the RC strains of Plasmodium berghei. METHODS: The concentrations of chloroquine in the plasma of normal mice and the mice infected with the N or the RC strains of P. berghei were analyzed by reverse phase HPLC. The pharmacokinetic parameters were measured with software 3P87. RESULTS: The t1/2 beta value was significantly lower in the mice infected with the RC strain than in normal mice and the mice infected with the N strain(P < 0.05), however, there were no significant differences between the mice infected with the N strain and normal mice. CONCLUSION: Elimination of chloroquine in the mice infected with the RC strain of P. berghei speed up significantly comparing with the mice infected with the N strain. PMID- 12567637 TI - [Effect of Nippostrongylus brasiliensis induced alterations in T helper cell subsets on Plasmodium berghei infection in mice]. AB - OBJECTIVE: To observe the anti-P. berghei ability of C57BL/6 mice after infected with Nippostrongylus brasiliensis alterations in T helper cell subsets in the course of Plasmodium infection, and the effect of the alteration on host's prognoses. METHODS: C57BL/6 mice were infected with Nippostrongylus brasiliensis subcutaneously, 3 wk later, the mice were injected with Plasmodium berghei intraperitoneally. The parasitemia was monitored daily. On days 0, 3 and 9, RNA from the spleens of infected mice was prepared for PT-PCR to analyse the changes of IFN-gamma and IL-4 mRNA during the infection course. RESULTS: Compared with control group, the peak-reaching time of parasitemia in the experiment group was delayed, and the bearing capacity of mice to malaria infection and the surviving time increased obviously. The IL-4 level in the experiment group was higher than that in the control group on day 0 of P. berghei infection, but all raised abnormally in both groups at the early stage of the infection; while IFN-gamma level in the experiment group was higher than that in the control group on day 3 after infection, and then began to reduce to some extent on day 9 of the infection in the experiment group. CONCLUSION: T helper cell subsets play an important role in antimalarial immunoregulation in mice. PMID- 12567638 TI - [Survey of pediculosis capitis in primary school pupils in the suburbs of Luoyang City]. PMID- 12567639 TI - [Observation on specific IgG4 antibody of schistosomiasis patients before and after treatment]. AB - OBJECTIVE: To observe the alteration of specific IgG4 antibody of schistosomiasis patients before and after treatment. METHODS: ELISA. RESULTS: The SEA-IgG4 and AWA-IgG4 positive rates of 27 schistosomiasis cases were 96.3% and 100%, respectively, their average OD values were 1.62 and 0.72. 6 months post treatment 18 cases were followed up, the positive rates were 94.4% and 100%, respectively, their average OD values were 1.06 and 0.56, respectively. 12 months post treatment all cases were followed up, the positive rates of SEA-IgG4 and AWA-IgG4 were 96.3% and 92.6%, respectively, their average OD values were 0.99 and 0.58, respectively. CONCLUSION: No obvious changes were found in the SEA-IgG4 and AWA IgG4 positive rates of 27 schistosomiasis cases before and after treatment, whereas the antibody level of specific IgG4 was decreased. PMID- 12567640 TI - [Ultra-pathological study on the syncytium of Schistosoma mansoni exposed to cyclosporin Ain vitro]. AB - OBJECTIVE: To study the ultra-pathological changes of syncytium of Schistosoma mansoni after cyclosporin A (CsA) treatment. METHODS: MF1 mice were infected with Schistosoma mansoni cercariae. Six weeks later, the adult worms were recovered by portal vein perfusion. After the worms were exposed to CsA of 20 micrograms/ml for 24 h, the drug-induced damage of the worm surface was observed by SEM and TEM. RESULTS: Incubation of male and female schistosomes with 20 micrograms/ml of CsA for 24 h resulted in disruption of the tegument and rupture of the spines. Progressive surface damage and swelling and vacuolization of the tegument led to eventual disruption of the syncytium. CONCLUSION: The antischistosomal action of CsA is direct, the syncytium is the main site for CsA attack. PMID- 12567641 TI - [Occurrence of Pagumogonimus veocularis in Fujian Province]. AB - OBJECTIVE: To prove that Fujian Province is also a natural focus of Pagumogonimus veocularis(Pv). METHODS: The adult worms were obtained from a cat fed with Pv metacercariae. RESULTS: Pv were found in Jianou, Fujian Province. All 1,873 Semisulcospira libertina showed negative. The positive rate of Tricula fujianensis and Erhaia jianouenesis were 0.10% (1/695) and 0.25% (5/2,038), respectively. The main crab host was S. fujianensis. Ps alone and mixed infection with Pv were found in the Sinopotamon, the infection rates were 36.8% (43/117) and 20.5% (24/117), respectively. The numbers of the metacercariae were 806 and 40, respectively. A cat was infected with 12 metacercriae of Pv, eggs were found in the stool 56 days after infection, and 6 worms were found in the lungs 68 days after infection. CONCLUSION: Fujian is one of the natural focus of Pv, cat is the adequate host. The fluke was identified as Pv according to the characteristics of the metacercariae. PMID- 12567642 TI - [Two cases of intestinal ascariasis]. PMID- 12567643 TI - [Laboratory diagnosis of Acanthamoeba keratitis]. AB - OBJECTIVE: To find a rapid method for diagnosing Acanthamoeba keratitis and identifying Acanthamoeba. METHODS: 10% potassium hydroxide (KOH) wet-mount preparations, Acanthamoeba culture, inverted phase contrast microscopy, and pathological examination using H. E. staining and PAS staining. RESULTS: Using corneal scrapings and corneal materials obtained from surgery, 7 cases and 5 cases of Acanthamoeba keratitis were diagnosed by 10% KOH wet-mount preparations. 6 strains of Acanthamoeba were isolated in corneal materials of 6 cases by protozoa culture method. The cysts, trophozoites and pseudopods on the trophozoites of Acanthamoeba were directly observed under the inverted phase contrast microscope. The cysts and trophozoites of Acanthamoeba were seen by H. E. staining and PAS staining with 20 h. CONCLUSION: Acanthamoeba keratitis could be rapidly diagnosed by 10% KOH wet-mount preparations and inverted phase contrast microscopy. Acanthamoeba organisms could be directly observed and identified under inverted phase contrast microscope. PMID- 12567644 TI - [Analysis of malaria situation in Qinglong County, Guizhou Province during 1992 1998]. PMID- 12567645 TI - [Acanthamoeba keratitis and its diagnosis, treatment and prevention]. PMID- 12567646 TI - [Effects of Lagenidium giganteum on the activities of three enzymes of Culex quinquefasciatus larvae]. AB - OBJECTIVE: To investigate the possible mechanism underlying the killing of Culex quinquefasciatus larvae by Lagenidium giganteum from the biochemical point of view. METHODS: The activities of alkaline phosphatase(AKP), acid phosphatase(ACP) and esterase(EST) were observed by using calcium-cobalt method, lead nitrate method and naphthol method. Results were photomicrographed and analysed quantitatively by image analysis using computer. RESULTS: The activities of AKP and ACP were decreased while the activities of EST were increased among the infected groups with different infection intensities. CONCLUSIONS: The changes of these enzyme activities might be one of the important mechanisms of action of Lagenidium giganteum against Culex quinquefasciafus larvae. PMID- 12567647 TI - Preliminary study on screening of egg mimotope of Schistosoma japonicum. PMID- 12567648 TI - [A case of falciparum malaria complicated with black water fever]. PMID- 12567649 TI - [Application of nuclear polyhedrosis virus of insects in parasitology]. PMID- 12567651 TI - [Detection of Demodex infection using squeezing-adhering combination method]. PMID- 12567650 TI - [Long-term effect of ICT application in monitoring filariasis control]. PMID- 12567652 TI - [A case with Cyclospora cayetanensis infection]. PMID- 12567653 TI - [Monitoring of sensitivity of artesunate-resistant Plasmodium falciparum to pyronaridine]. PMID- 12567655 TI - [Views on a non-endemic area for filariasis in Yunan Province]. PMID- 12567654 TI - [Inducible expression of MSP1 gene of Plasmodium falciparum by a tetracycline controlled promoter]. AB - OBJECTIVE: To express the entire MSP1 gene of Plasmodium falciparum and its C terminal 42 kDa fragment using a tetracycline-controlled PLtetO-1 promoter. METHODS: The entire MSP1 gene and 42 kDa fragment gene were cloned into the plasmid of pZE11, and transformed into E. coli DH5 alpha Z1. Restriction enzyme analysis, SDS-PAGE and Western blotting were used to examine two recombinant plasmids and their expression in E. coli DH5 alpha Z1. RESULTS: The recombinant plasmids of pZE11/MSP1 and pZE11/MSP1-42 were constructed successfully. The expressive products about 190 kDa and 42 kDa of two genes in E. coli DH5 alpha Z1 were identified by SDS-PAGE and Western blotting. CONCLUSION: Tightly controlling expression of the MSP1 gene in E. coli is essential to reduce the toxicity of the product to its host cells as well as to provide a feasibility to construct Salmonella vaccine strain which can inducibly express the important malarial vaccine candidate gene. PMID- 12567656 TI - [Effect of anti-mosquito-midgut antibodies on the development of oocysts of Plasmodium yoelii in Anopheles stephensi]. AB - OBJECTIVE: To study the effect of anti-mosquito-midgut antibodies on the development of P. yoelii in A. stephensi. METHODS: Ookinetes and oocysts in midgut and sporozoites in the salivary glands of mosquitoes were examined respectively at 14 h and 9 d and 12 d after the mosquitoes ingested infected blood containing anti-midgut antibodies by membrane feeders. The effect of anti midgut antibody level and repeated feedings on the development of the oocysts in the mosquitoes was also observed. RESULTS: The anti-midgut antibodies reduced evidently not only the infection rates of the oocysts and the number of the sporozoites in salivary gland but also infection degree of ookinetes and the index of oocysts in the mosquitoes compared with control group (P < 0.05). The higher the antibody level, the more the times for mosquitoes digested antibodies, and the fewer the number of oocysts. CONCLUSION: The anti-mosquito-mindgut antibodies inhibited ookinetes and oocysts of P. yoelii in A. stephensi, with oocysts in particular. PMID- 12567657 TI - [Plasmodium falciparum: antimalarial activities of antisense oligodeoxyribonucleotides targeted to histidine-rich protein genes in vitro]. AB - OBJECTIVE: To study the inhibition of growth of Plasmodium falciparum cultured in vitro with antisense oligodeoxyribonucleotide (AS ODN) against histidine-rich proteins (HRP). METHODS: The ODNs against HRP II and HRP III were synthesized and used to study the antimalarial activities in vitro. Plasmodium falciparum (FCC1/HN strain, China) were exposed to AS DONs for 48 h, and the growth inhibition was determined by microscopic examination. RESULTS: At 1 mumol/L, all ODNs inhibited parasite growth and development in a target-independent manner. However, when the ODN concentrations were between 0.01 and 0.5 mumol/L, the AS ODN significantly inhibited the growth and development of P. falciparum compared with ODN controls (P < 0.01). Inhibition by the sense strand ODN did not differ significantly from the control group (P > 0.05). CONCLUSION: Blockade of the expression of HRP II and HRP III AS ODN could inhibit P. falciparum cultured in vitro. PMID- 12567658 TI - [A case with Ancylostoma duodenale larvae in ascitic acid]. PMID- 12567659 TI - The schizontocidal activity of daphnetin against malaria parasites in vitro and in vivo. AB - OBJECTIVE: To investigate the in vitro and in vivo schizontocidal activity of daphnetin. METHODS: Schizontocidal activity of daphnetin was tested using an in vitro assay based on the routine in vitro cultivation of P. falciparum FCC1 strain. The in vivo antimalarial effects of daphnetin at various dosages were assessed in mice infected with P. b. erghei ANKA according to "4-day suppress assay". RESULTS: In vitro, daphnetin exhibited potent schizontocidal activity comparable to chloroquine(CQ) at the dose range of 1-10 mumol/L. In vivo, 50 or 100 mg/kg.d-1 x 4 d daphnetin i.g. and 10, 50 or 100 mg/kg.d-1 x 4 d dephnetin i.p. showed antimalarial efficacy comparable to CQ 10 mg/kg.d-1 x 4 d i.g. in mice infected with P. berghei ANKA, evaluated by both the reduction rate of parasitemia on D4 and the average surviving days in 30 days. CONCLUSION: Daphnetin displays certain schizontocidal activity both in vitro and in vivo. PMID- 12567660 TI - The role of CD8+ T cells in the pathogenesis of Pneumocystis carinii pneumonia in mice depleted of CD4+ T cells. AB - OBJECTIVE: To define the cell populations contributing to inflammation-associated respiratory impairment in Pneumocystis carinii pneumonia (PCP). METHODS: The host inflammation response was observed in CD4+ T cell-depleted mice and in CD4+ and CD8+ T cell-depleted mice infected with P. carinii via intratracheal inoculation. RESULTS: Mice depleted of both CD4+ and CD8+ T cells developed infection with neither increased respiratory rate nor lung injury. In contrast, mice depleted of CD4+ T cells alone exhibited severe pulmonary inflammation, and increased respiratory rates. Respiratory compromise was associated with the presence of activated CD8+ cells and neutrophils in the BALF. CONCLUSION: The host's inflammatory cell response to P. carinii directly impairs pulmonary function and contributes to the pathogenesis of PCP, CD8+ T cells appear to play a major role. PMID- 12567661 TI - Gene sequencing for identification of Paragonimus eggs from a human case. AB - OBJECTIVE: To identify the etiologic agent from a paragonimiasis patient using molecular techniques. METHODS: The complete nuclear ribosomal DNA second internal transcribed spacer (ITS2) gene sequence of eggs in sputum from a paragonimiasis patient was obtained by directly auto-sequencing its PCR product. ITS2 genes from eggs of Paragonimus westermani and Pagumogonimus skrjabini (both from animal hosts) were also sequenced for comparison. In addition, morphological comparisons were made with the eggs of the two species. RESULTS: The ITS2 gene from the human case was 100% identical with the sequence from the eggs of P. westermani from an experimentally infected dog but only 92% identical with the sequence from the eggs of P. skrjabini. Morphologically, the eggs from the human case more resembled those from P. westermani infected dog. CONCLUSION: The patient was diagnosed to be suffered from paragonimiasis westermani by gene sequence analysis. PMID- 12567662 TI - [Analysis of the infection mode of 61 cases with taeniasis in Jilin City]. PMID- 12567663 TI - [Localization and characterization of partial immunodominant antigen epitopes of Trichinella spiralis]. AB - OBJECTIVE: To screen and characterize immunodominant antigen epitopes on the soluble antigens of Trichinella spiralis (T.s.). METHODS: 15 monoclonal antibodies (McAbs) against T.s. muscle larva(ML) soluble antigens were obtained by using hybridoma technique. The reactivity of monoclonal and polyclonal antibodies were tested by ELISA, Western blotting and indirect immunofluorescence assay(IFA). RESULTS: The Western blotting result showed that of the 15 McAbs, 6 could bind to the T.s. ML antigens displaying molecular weights of 40-70 kDa. Polyclonal sera could react with more than 10 bands having molecular weights of 20-200 kDa. Among the 6 McAbs, 4 could recognize epitopes on the cuticle surface and the other two could recognize epitopes on both the cuticle surface and the stichosome. CONCLUSION: The antigen epitopes of T.s. recognized by 6 McAbs had been characterized. PMID- 12567664 TI - [Gene cloning and characterization of mitochondria-related protein of Schistosoma japonicum]. AB - OBJECTIVE: To subclone and characterize a cDNA clone coding for Schistosoma japonicum (S.j.) mitochondria-related protein. METHODS: The open reading frame of the fragment(Sj338/24) obtained from an adult worm cDNA library of S.j. was analysed, at the upstream and downstream of the open reading frame(ORF) the primers A and B were designed, respectively, and the cDNA fragment was used as PCR template. The Sj338 gene fragment obtained was amplified by PCR method and then subcloned into pGEM-T vector for sequencing. The gene sequence was analyzed and the target fragment was restrictedly digested and subcloned into expression vector pGEX-6P-1. The expressed recombinant protein was purified and characterized. RESULTS: The cloned Sj338 gene was demonstrated to be 487 bp long containing one 459 bp ORF, encoding a protein with a molecular weight of 17 kDa. The nucleotide sequence of the cloned gene Sj338 had higher homology with those genes coding for mitochondrial outer membrane protein of Homo sapiens and Rattus norvegicus. The recombinant construct of pGEX-6P-1/Sj338 could be expressed efficiently and the antigenicity of its product rSj338 has been demonstrated by Western blotting. CONCLUSION: Sj338 may be the gene coding for S.j. mitochondria related protein and the recombinant protein may be used as a new vaccine candidate. PMID- 12567665 TI - [Clinical analysis of 41 cases with eosinophil asthma associated Ascaris infection]. PMID- 12567666 TI - [Investigation of anti-Toxoplasma gondii antibodies in immunodeficient patients]. AB - OBJECTIVE: To investigate the presence of anti-Taxoplasma gondii antibodies in immunodeficient patients. METHODS: T. gondii-specific immunoglobulin G (IgG) antibodies in serum samples from 371 immunodeficient patients were detected by enzyme-linked immunosorbent assay (ELISA). The patients were with solid malignancies (including untreated digestive system malignancies and solid malignancies received chemotherapy), chronic liver diseases, patients received immunosuppressant therapy (dermatomyositis, psoriasis, pemphigus, post-renal transplantation, systemic lupus erythematosus and other miscellanies), lymphoma, leukemia and diabetes. 100 normal serum samples served as controls. RESULTS: The positive rate of patients with solid malignancies received chemotherapy, solid malignancies received chemotherapy, chronic liver diseases, systemic lupus erythematosus and leukemia was 19.0%, 33.3%, 16.5%, 45.4% and 20.0%, respectively, being significantly higher than that of the control group(P < 0.05). CONCLUSION: The immunosuppressed patients are highly predisposing to secondary T. gondii infection. PMID- 12567668 TI - [Establishment of an animal model of Capillaria hepatica]. AB - OBJECTIVE: To establish an animal model of Capillaria hepatica. METHOD: Sixteen rats and two cats were infected with the embryo eggs of Capillaria hepatica through mouth. RESULTS: Of 16 infected rats, 2 were negative, 14 were positive with Capillaria hepatica. 2 cats were negative. CONCLUSION: Rats could be used as an animal model of Capillaria hepatica. PMID- 12567667 TI - [Comparison of circulating antigen detection in schistosomiasis japonica patients using two methods of ELISA]. AB - OBJECTIVE: To compare 2 kinds of ELISA methods for detecting circulating antigen in schistosomiasis patients. METHODS: Monoclonal antibody-based dot-ELISA and sandwich ELISA. RESULTS AND CONCLUSION: The positive rates of dot-ELISA were higher than or similar to those of sandwich ELISA, the former being 48.0%-98.8% and the latter being 12.0%-93.8%. The specificities were 95.4%-100% and 90.0%, respectively. PMID- 12567669 TI - [Image analysis of the Schistosoma juponicum eggs]. PMID- 12567670 TI - [Investigation on Schistosoma japonicum infection in rodents in Yunnan Province]. AB - OBJECTIVE: To investigate the role of rodents in the transmission of schistosomiasis in endemic areas in Yunnan Province. METHODS: Kato-Katz method was used to study the infection rate of rodents, humans and domestic animals. RESULTS: The infection rate was 0.9% (32/3,411) for rodents, 15.6% (461/2,964) for humans, and 9.6% (239/2,482), for domestic animals. The EPG(mean) was 5.8 for rodents, 1.8 for humans, 0.1 for cattle, 0.1 for horses and 0.02 for pigs. CONCLUSION: The rodents played a minor role in the transmission of schistosomiasis in the plateau area of Yunnan province. PMID- 12567671 TI - [Progress in the research on genetic transfection of malaria parasites]. PMID- 12567672 TI - [Progress in the research on the tachyzoite-bradyzoite conversion of Toxoplasma gondii]. PMID- 12567673 TI - [Progress in the research on recombinant antigens for diagnosing schistosomiasis]. PMID- 12567674 TI - [Pathological charges induced by the growth of Pneumocystis carinii in rat lungs]. PMID- 12567675 TI - [Analysis of the G- and C-banding of chromosome of larval Echinococcus granulosus from humans]. PMID- 12567676 TI - [Preliminary study on a proteolytic enzyme from Cysticercus cellulosae]. PMID- 12567677 TI - [A staining method for defecting microfilaria in preserved hemolysed smear preparations]. PMID- 12567678 TI - [Diagnosis of kala-azar by rK39 dipstick assay]. PMID- 12567680 TI - [Follow-up survey of chronic and advanced schistosomiasis patients in Qingpu District, Shanghai]. PMID- 12567679 TI - [Epidemiological survey of malaria in Nangiao endemic area of Hunan Province using IFAT and PCR]. PMID- 12567681 TI - [Evaluation of the effects of technology and economy of different plans to control schistosomiasis]. PMID- 12567682 TI - [Decystation and migration of metacercariae of Pagumogonimus skrjabini]. PMID- 12567683 TI - [Malaria situation in the People's Republic of China in 1999]. AB - Although the middle and lower reaches of Changjiang River were consecutively stricken by severe flood in 1998 and 1999, the transmission of malaria was not frequent and prevalence of the disease was basically stable with no reports of outbreaks in the above areas, which was attributed to the intensification of malaria surveillance, prompt implementation of integrated measures including mosquito control and chemoprophylaxis, as well as the zoophilous trend of Anopheles sinensis, the vector in the stricken areas. According to the case reporting system established on the basis of professional institutions of 22 provinces, autonomous regions and municipalities (P/A/M), the number of malaria cases in the country totalled 29,039 in 1999, with lethal cases of 67. Based on pilot-site surveillance and investigation of some localities, the actual number of malaria cases was estimated to be 250,000-300,000 in 1999. Hainan and Yunnan are still the major malarious provinces. The elongated borderline and increase in migratory population contributes to the difficulties in malaria control in Yunnan, consequently, the incidence of malaria was progressively upgrading in the last three years, the reported number of falciparum malaria cases and deaths was markedly increased in 1999 as compared with that in 1998, exhibiting a crucial status of prevalence in the province. In Hainan, circa 80% of malaria cases were infected via transmission by An. dirus away from villages, hence difficulties existed in malaria control; in areas affected by An. anthropophagus where a population of more than 100 million resided, relatively high incidence of malaria was noted, the prevalence was unstable, sometimes focal outbreaks occurred, and incidence of 20% was reported in a few villages and townships; in area where the only vector was An. sinensis, the prevalence was rather stable, the incidence of malaria was decreased to < 0.1 @1000 in most places. A total of 26,797 people proved to be positive for malaria parasite in the process of blood examination on febrile cases of 7,816,182 man/time in area where lived 691 million population, covering 1,336 counties in 19 P/A/M, the mean parasite rate being 0.34%, however, the rate was as high as 2.09% in migratory people. Blood examination on targetted people including the inhabitants in the epidemic foci, children and migratory population for 362,512 man/time detected 1,781 malaria parasite carriers, with a mean parasite rate of 0.49%. Prevalence of falciparum malaria was still confined to 44 counties of Hainan and Yunnan provinces. Imported falciparum malaria cases were found in 114 counties of 16 P/A/M (areas not endemic for falciparum malaria in Hainan and Yunnan also included). Altogether 5,466 cases and 227 carriers of falciparum malaria (including mixed infection of Plasmodium vivax and Plasmodium falciparum) were detected this year. PMID- 12567684 TI - [Human parasite antigen-specific IFN-gamma response in schistosomitsis japonica endemic area]. AB - OBJECTIVE: To explore the characteristics of human schistosome antigen-specific IFN-gamma response in a population in an area endemic for schistosomiasis japonica. METHODS: Three neighboring villages were chosen on Nanshan Island of Poyang Lake. 65 egg-negative persons and 64 egg-positive ones were selected randomly from the residents aged 14-41 years according to the egg counts by Kato Katz thick smear method. IFN-gamma was measured in the whole blood culture supernatant after stimulated by the schistosome soluble egg antigen (SEA) and soluble worm antigen preparations (SWAP). Serum isotype-restricted antibody was detected by ELISA. RESULTS: IFN-gamma levels induced by both SEA and SWAP were increased significantly after praziquantel treatment. The SEA-specific IFN-gamma level of the uninfected group was much higher than that of the reinfected group. A negative correlation between IFN-gamma level and IgG4 production was found, reflecting that IFN-gamma might be associated with the resistance to schistosome reinfection. CONCLUSION: The changes in IFN-gamma level might play an important role in association with the resistance to schistosome reinfection. PMID- 12567685 TI - [Purification of soluble egg antigen (SEA) 38 kDa molecule of Schistosoma japonicum and preliminary evaluation of its immunodiagnostic efficacy]. AB - OBJECTIVE: To study the applicability of soluble egg antigen (SEA) 38 kDa molecule of Schistosoma japonicum in the immunodiagnosis of schistosomiasis. METHODS: The target immunodiagnositic antigen SEA 38 kDa molecule was separated and purified by SDS-PAGE and electroelution technique and evaluated by SDS-PAGE and Western blotting. To compare their immunodiagnostic efficacy, the purified SEA 38 kDa and SEA were used to examine the sera from 182 cases including 31 acute schistosomiasis patients, 31 chronic schistosomiasis patients, 60 normal human controls, 30 paragonimiasis patients and 30 clonorchiasis patients. RESULTS: The positive rates of purified SEA 38 kDa and SEA for acute schistosomiasis patients, chronic schistosomiasis patients, normal human controls, paragonimiasis patients and clonorchiasis patients were 90.3%, 90.3%, 1.7%, 0, 0 and 90.3%, 83.9%, 3.3%, 0, 0, respectively. The P/N ratio of purified SEA 38 kDa and SEA in immunodiagnosis of schistosomiasis by ELISA had been compared by two-sample t-test. It was demonstrated that there existed significant differences between purified SEA 38 kDa and SEA in the three groups of acute and chronic schistosomiasis patients and normal persons(P < 0.001). The P/N ratio of SEA 38 kDa in detecting the sera from schistosomiasis patients was obviously higher than that of SEA, but when SEA 38 kDa was used to examine normal human sera, the result was converse. CONCLUSION: The purified SEA 38 kDa has higher sensitivity and specificity and immunodiagnostic potential. PMID- 12567686 TI - [Role of TNF-alpha and ICAM-1 in pathogenesis of cerebral malaria]. AB - OBJECTIVE: To investigate the role of TNF-alpha and ICAM-1 in the pathogenesis of cerebral malaria. METHODS: Immunohistochemical method and ELISA were employed to examine the expression of ICAM-1 on the brain microvessel endothelium and to detect the production of serum TNF-alpha in P. berghei ANKA strain infected-CBA/J mice. RESULTS: Serum TNF-alpha levels of mice were apparently higher and the ICAM 1 expression was more evident in P. berghei ANKA infected-CBA/J mice than in control groups. rTNF-alpha i.p. injection could enhance the development of CM and the expression of ICAM-1 on brain endothelial cells(EC). CONCLUSION: Excessive production of TNF-alpha may mediate the expression of ICAM-1 on brain EC and hence cause the development of CM. PMID- 12567687 TI - [Molecular phylogeny of Giardia lamblia based on triose phosphate isomerase (tim) gene sequence]. AB - OBJECTIVE: To investigate the intraspecies phylogeny and genetic diversity of G. lamblia. METHODS: Total genomic DNA was extracted and partial fragments of tim gene were amplified by PCR. All the nucleotide sequences were analyzed by means of phylogenetic analysis, which were constructed with pasimony and N-J methods. RESULTS: A total of 124 variable sites (23% of the total sequences detected) were defined, most of which were found at the silent sites of codons. Two similar phylogenetic trees were constructed, subdividing 16 isolates into two groups. CONCLUSION: The genetic diversity of G. lamblia appear to be little affected by both the host and geography, while natural-selection might play an important role at the DNA molecular evolution level of tim gene. The tim gene may be considered to be a very useful genetic marker of the population genetic structure of G. lamblia. PMID- 12567688 TI - Relationship of the livestock trade to schistosomiasis transmission in mountainous area. AB - OBJECTIVE: To study the relationship between the livestock trade and schistosomiasis transmission and to provide an evidence for making a strategy of schistosomiasis control in mountainous areas. METHODS: A retrospective survey and analysis was conducted to investigate the prevalence of schistosomiasis in both humans and livestock (cattle, horses, mules, donkeys and pigs), and the number and migration of livestock in Weishan County, Yunnan Province in 1980-1991. RESULTS: A positive correlation was found between the infection rate of residents and the numbers of livestock migration (R = 0.9151, P < 0.01). During 1980 to 1991 the infection rate was increased gradually along with the development of livestock husbandry, especially, from the economic reforms since 1984. In 1984 there was positive correlation in the infection rate both human and livestock (R = 0.8458, P < 0.05). The results show that the infection rates of livestock on sale including cattle, horses, mules, donkeys and pigs are 9.54%, 29.39%, 16.38%, 14.47%, 25.73% and 11.11%, respectively. CONCLUSION: The infection rate of human and livestock arises by parallel. The high frequency of livestock trade resulted in serious spreading of the infection source of schistosomiasis. The migration of the infected livestock might be an important factor in transmitting schistosomiasis. PMID- 12567689 TI - [Survey of susceptibility of anopheline vectors to insecticides in a malaria mesoendemic area, Hubei Province]. AB - OBJECTIVE: To determine the susceptibility of Anopheles anthropophagus and Anopheles sinensis to deltamethrin and DDT in an area where different counter measures had been adopted. METHODS: The mortality of the vectors was examined by using the standardised WHO test. RESULTS: The mortality for tentative diagnostic dose in An. anthropophagus to deltamethrin were 83.8%, 83.7% and 84.7%, respectively in the areas where impregnated net had been used for 1 year or 3 years and DDT residual spraying had been carried out for 3 years, suggesting that An. anthropophagus was an initially resistant group. The LT50 were 8.69, 7.48 and 9.87 min, respectively. The mortality for tentative diagnostic dose in An. sinensis to deltamethrin and DDT were 76.5%, 57.0% and 79.0%, respectively in three survey areas, suggesting that An. sinensis was a resistant group. The LT50 were 12.0, 15.4 and 11.2 min, respectively. The mortality for tentative diagnostic dose in An. sinensis to DDT was 95.8% in impregnated net area, suggesting that An. anthropophagus was an initially resistant group. The LC50 was 0.73%. The mortality for tentative diagnostic dose in An. sinensis to DDT was 44%, suggesting that An. sinensis was a highly resistant group. The LC50 was more than 4%. CONCLUSION: A substantial use of insecticides in the rice paddy field has resulted in resistance of An. sinensis, whereas no apparent resistance of An. anthropophgus has been found. PMID- 12567690 TI - [A case of kala-azar in Mianyang City]. PMID- 12567691 TI - [Detection of specific antibodies in saliva of patients with chronic schistosomiasis japonica]. AB - OBJECTIVE: To investigate the feasibility of detecting specific antibodies in the saliva of schistosomiasis patients. METHODS: Specific antibodies in saliva samples of 32 schistosomiasis patients and 140 normal individuals were detected by using ELISA and the results were compared with those detected for specific antibodies in serum. RESULTS: The sensitivity and specificity of the ELISA were 90.6% and 94.4%, respectively, being slightly lower than that of the 100% sensitivity and 96.2% specificity for the serum. No significant difference between them was found (P > 0.05). CONCLUSION: Detecting specific antibodies in saliva can be used for immunodiagnosis of schistosomiasis japonica as a noninvasive method in field surveys. PMID- 12567692 TI - [Report of 8 cases of angiostrongyliasis cantonensis]. PMID- 12567693 TI - Rapid screening with a recombinant antigen (rK39) for diagnosis of visceral leishmaniasis using dipstick. AB - OBJECTIVE: To evaluate the diagnostic value of the recombinant antigen of 39 amino acid repeats encoded by a kinesin-like gene of Leishmania changasi (rK39) in serodiagnosis of visceral leishmaniasis (VL). METHODS: In Kashi, Xinjiang, 13 VL patients with splenomegaly and bone marrow aspirate culture positive were subjected to dipstick assay. A drop of whole blood or serum from patient was placed at the absorbing pad at the bottom of the dipstick. Flooding of the bottom protein with buffer allows serum proteins to migrate upwards, producing the positive band and Western blot analysis of rK39 subsequently performed with the sera collected. RESULTS: The end-point titers of anti-rK39 antibodies of these sera were determined by ELISA and found to fall within the range of 10(-2) to 10( 4), which were consistent with the intensity of their reaction with rK39 in dipstick assay. The positive sera could also recognize the specific rK39 band as analyzed by Western blot analysis. CONCLUSION: The rK39 dipstick assay is more rapid, specific, sensitive and less invasive than the conventional methods of diagnosis for VL in the areas of low endemicity. PMID- 12567694 TI - [Clinical comparative trial of co-artemether and benflumetol (two formulations) in the treatment of falciparum malaria]. AB - OBJECTIVE: To observe the efficacy and safety of co-artemether and one of its components benflumetol (two formulations) in the treatment of patients with falciparum malaria. METHODS: Adopting double-blining, randomization and comparative method, all patients were hospitalized and observed for 28 days after treatment. RESULTS: Of 150 patients, 51 patients were treated with co-artemether group(A), 50-patients were treated with benflumetol tablet group(B), 49 patients were freated with benflumetol capsule group(C). The mean fever clearance times for groups A, B and C were 17.1 +/- 8.6, 34.0 +/- 23.2 and 29.4 +/- 24.9 hours, respectively; the mean parasite clearance times were 29.7 +/- 8.9, 51.6 +/- 14.1 and 54.7 +/- 17.4 hours respectively; the cure rates in 28 days for groups A, B and C were 98.2%, 92.0% and 95.8%, respectively. No apparent side-effect was observed. CONCLUSION: Co-artemether and benflumetol (2 formulations) are effective for the treatment of patients with falciparum malaria but co-artemether is more effective than benflumetol (2 formulations) in terms of controlling symptoms and killing parasites. PMID- 12567695 TI - [A case of dermatitis caused by Paederus fuscipes]. PMID- 12567697 TI - [A case of strongyloidiasis]. PMID- 12567696 TI - [Effect of artemether on enzymes involved in carbohydrate metabolism of Schistosoma japonicum]. AB - OBJECTIVE: To study the effect of artemether on several enzymes involved in carbohydrate metabolism of Schistosoma japonicum. METHODS: Mice infected with Schistosoma japonicum cercariae for 4-5 weeks were administered intragastrically with artemether 300 mg/kg and killed 24-72 h after medication. The supernatant fluids of female and male worm homogenates were prepared for determining 9 essential enzymes of carbohydrate metabolism by using horizontal starchgel electrophoresis. RESULTS: The activities of 8 out of 9 enzymes (i.e. hexokinase, aldolase, glucosephophate isomerase, malate dehydrogenase, malic enzyme, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and mannose-6 phosphate isomerase) in female worms from artemether-treated mice were obviously inhibited 24-72 h after treatment. In male worms, only aldolase, mannose-6 phosphate isomerase and glucose 6-phosphate dehydrogenase were slightly inhibited. CONCLUSION: Artemether displayed apparent effects on the carbohydrate metabolism of female schistosomes. PMID- 12567698 TI - [Detection of Toxoplasma infection in women with gynaecologic neoplasms using ELISA]. AB - OBJECTIVE: To understand the concurrent in Toxoplasma infection among women with cervical cancer. METHODS: The anti-Toxoplasma IgG, IgM antibodies (Abs) and circulating antigens (CAg) in 247 sera (from 50 cases with cervical cancer in group 1, 59 cases with gynaecological benign tumors in group 2 and 138 cases with other diseases in group 3) were detected by ELISA. RESULTS: The total positive rates of the CAg and the Abs in the three groups were 44.0% (22/50), 25.4% (15/59) and 20.3% (28/138), respectively. The positive rates were 20.0%, 11.9% and 10.9%, respectively for IgG 20.0%, 13.6% and 8.7%, respectively for IgM and 14.0%, 5.1% and 1.4% respectively for CAg. CONCLUSION: The Toxoplasma infection rate of the cervical cancer group was higher than those of the other two groups, particularly the positive rates of CAg and IgM. PMID- 12567699 TI - [Observation on the persistence period and transmission of residual microfilaremia with medium and higher density]. AB - OBJECTIVE: To explore the persistence period of medium and higher density of microfilaremia and its role in the transmission of filariasis in areas with filariasis basically eradicated. METHODS: The residual microfilaremia was followed up and the population were regularly examined by thick blood film assay. Culex quinquefaciatus were dissected to determine the natural infection rate and density of filarial larvae. The filarial antibody levels were detected by using IFAT. RESULTS: Two cases with medium and higher density of microfilaremia were found to be microfilaremia positive for twelve years. The natural infection rate and density of filarial larvae in Culex quinquefasciatus dropped down year by year, one case of new microfilaremia was found in the sixteenth year. CONCLUSION: The persistence period of residual medium and higher density of microfilaremia lasted for more than twelve years. The individual residual medium and higher density of microfilaremia cases still possess the potential for transmission of filariasis. PMID- 12567700 TI - [Evaluation of prevalent status of kala-azar in endemic area of Hubei Province]. AB - OBJECTIVE: To identify the diagnosis of 14 cases with "kala-azar" previously reported in Hubei Province since 1989 and to evaluate the present status of the disease. METHODS: Bone marrow smear, immunological test (LDTand IFAT), mouse inoculation, fungi culture, vector (Phlebotomus) and reservoir host (dog) survey. RESULTS: Among the 14 cases, 12 cases were progressive disseminated histoplasmois, one was an imported case and another one was a remnant skin type "kala-azar" case. An immunological survey was conducted during 1997-1999 in 25 townships located in 23 counties of 9 prefectures and cities. A total of 13,890 persons had been examined by LDT, the positive rate was 0.07%, no positive case was found among the persons under 40 years of age. During the survey, 1,756 persons had been examined by IFAT, the results were negative. No Phlebotomus chinensis had been found in the surveyed area. A total of 302 dogs were examined, no sick dog had been found. CONCLUSION: Kala-azar has been eradicated in Hubei Province. PMID- 12567701 TI - Effect of Schistosoma japonicum infection on serum testosterone levels in mice. AB - OBJECTIVE: To determine the effect of Schistosoma japonicum infection on the testoterone level in the sera from male C57BL/6 mice. METHODS: Radioimmunoassay was used to examine testosterone levels in sera of 9 male mice experimentally infected with Schistosoma japonicum. RESULTS: The serum testosterone levels reduced significantly in all experimentally infected animals 45 days after infection, as compared with the uninfected controls. CONCLUSION: Infection with Schistosoma japonicum decreases testosterone levels in the mouse host. PMID- 12567702 TI - [Survey of an outbreak of imported paragonimiasis]. PMID- 12567703 TI - [Epidemiological investigation on an outbreak of angiostrongyliasis cantonensis in Wenzhou]. AB - OBJECTIVE: To find out the cause and transmission factors of an outbreak of eosinophilic meningoradiculitis in Wenzhou City during October to November in 1997. METHODS: In addition to a retrospective cohort study on 182 persons who had dined at a same restaurant of the city, etiological and immunoserological investigations were conducted. RESULTS: Among 105 cases who had eaten the undercooked Ampullaria gigas in that restaurant, 47 cases were infected, the incidence rate was 44.8%; whereas none of the persons who did not eat the under cooked Ampullaria gigas in that restaurant got the infection, the difference between the two groups being significant(P < 0.001). The incidence rate of the cases who had eaten 4 and more pieces of the undercooked Ampullaria gigas was 86.2%(25/29), while that of those who had eaten less than 4 pieces was 28.9%(22/76), the difference between the two being very significant (chi 2 = 27.83, P < 0.001, RR = 2.98). The third stage larvae of the Angiostrongylus cantonensis were found out from Ampullaria gigas living in the same locality, and the adult worms of Angiostrongylus contonensis were found out from the rats living in the locality else. The positive rate of serum anti-Angiostrongylus cantonensis antibody in the patients was 84.0%. CONCLUSION: On the basis of the clinical, epidemiological, immunoserological and etiological investigations, an outbreak of the angiostrongyliasis cantonensis caused by eating undercooked Ampullaria gigas was confirmed. PMID- 12567705 TI - [Progress in the research on echinococcosis control]. PMID- 12567704 TI - [Report of a case with Schistosoma mansoni egg in the urine]. PMID- 12567706 TI - [Endemic status of hookworm disease and progress in vaccine preparation]. PMID- 12567707 TI - [11 cases of hydatid disease of the common bile duct]. PMID- 12567708 TI - [Toxic effects of levamisole]. PMID- 12567709 TI - [Report of a case with albendazole-resistant Strongyloides stercoralis infection]. PMID- 12567710 TI - [Evaluation of the effect of 40 years anti-malaria measure in Henan Province]. PMID- 12567711 TI - [Analysis of the focal surveillance of filariasis in Jiangxi Province]. PMID- 12567712 TI - [Field trial of snail control using membrane-covering method in mountainous area]. PMID- 12567713 TI - [Surveillance report of basical eradication of filariasis in Zoucheng City, Shandong Province]. PMID- 12567714 TI - [Comparison of rDNA-ITS2 sequences and morphological characters of Anopheles kunmingensis and Anopheles langshanensis in China, with discussion on taxonomic status]. AB - OBJECTIVE: To determine the taxonomic status of Anopheles kunmingensis (AK) and An. liangshanensis(AL) from China. METHODS: Sequence differences of rDNA-ITS2 and main morphological characters variability between different sources of AK and AL were compared. RESULTS: The level of rDNA-ITS2 sequence differences among eight samples was under 2.9%. The total occurrence rates of main morphological characters examined in the female mosquitoes with pale fringe spot on V5.2, white basal band (or spot) on hind tarsomere IV, and larvae with bifurcated head hair 2 C were 43%(9/21), 89%(17/19), 40(4/10) in AK, and 79%(34/43), 44%(17/39), 20%(4/20) in AL, respectively. These rates calculated from separate colonies fluctuated within a wide range and overlapped, suggesting that there was no definite, stable morphological difference between AK and AL. CONCLUSION: The morphological and molecular variations between AK and AL were small and belong to intraspecific level. The AK and AL may be considered as the same species, and that the An. kunmingensis may be recognized as the synonym of An. liangshanensis. PMID- 12567715 TI - [Clinical analysis of 10 cases with paragonimiasis westermani]. PMID- 12567717 TI - [A case with Taenia saginata eggs in cerebrospinal fluid]. PMID- 12567716 TI - [Evaluation of Schistosoma japonicum tegumental membrane antigen (SjMAg) in immunodiagnosis and efficacy assessment]. AB - OBJECTIVE: To evaluate the specificity and sensitivity of SjMAg in detecting specific antibodies in sera of patients with schistosomiasis japonica and in assessing therapeutic efficacy. METHODS: SjMAg-ELISA was used to determine the specific IgG and IgG4 in sera of patients with schistosomiasis japonica at different time points after chemotherapy. RESULTS: SjMAg-ELISA was as sensitive and specific as SEA-ELISA. The negative conversion rate of IgG detected by SjMAg ELISA 12 months after treatment was 80.0%, being significantly higher than 43.3% by SEA-ELISA. The negative conversion rate of IgG4 detected by SjMAg-ELISA 12 months after treatment was 93.3%, being significantly higher than 60.0% by SEA ELISA. The negative conversion rates of IgG and IgG4 2 yr after treatment were 92.9% and 97.6% respectively. CONCLUSION: SjMAg-ELISA is comparable to SEA-ELISA in diagnosing schistosomiasis japonica and is more effective in assessing therapeutic efficacy. PMID- 12567718 TI - [Cloning of Cysticercus cellulosae AgB cDNA coding region]. AB - OBJECTIVE: To amplify and clone Cysticercus cellulosae AgB cDNA coding region. METHODS: The AgB cDNA was amplified by RT-PCR technique from the total RNA of Cysticercus cellulosae. It was cloned into the vector pUC118 and sequenced. RESULTS: The PCR amplified product was a single band of 2.6 kb in size. The sequences of AgB cDNA coding region has 99.8% homology with that of Australian Cysticercus cellulosae, and their amino acid sequences have 99.3% homology. CONCLUSION: Cysticercus cellulosae AgB cDNA coding region has been cloned successfully. PMID- 12567719 TI - [The synergistic action of guanghuoxiang volatile oil and sodium artesunate against Plasmodium berghei and reversal of SA-resistant Plasmodium berghei]. AB - OBJECTIVE: To study the synergistic action of a combination of guanghuoxiang volatile oil (B) and sodium artesunate (SA) against Plasmodium berghei (P. b) and the resistance-reversal activity against SA-resistant P. b (P. b SA-R). METHODS: Mice infected with P. b N or P. b R were treated with a combination of B and SA respectively by 4-day suppressive test method and linear regression to calculate the SD50 of B and SA for each drug alone and in combination (equally effective dose compatibility). RESULTS: B alone, N:SD50 = 87.64 +/- 19.58(GKD), R:SD50 = 43.24 +/- 7.71(GKD); SA alone, N:SD50 = 0.88 +/- 0.01(MGKD), R:SD50 = 27.69 +/- 0.93(MGKD). B and SA combination, N:B SD50 = 36.89 +/- 4.57(GKD), SA SD50 = 0.39 +/- 0.05 (MGKD); R:B SD50 = 7.40 +/- 1.30(GKD), SA SD50 = 4.21 +/- 0.74(MGKD). The synergistic indexes of B and SA in combination were 2.2 for N and 6.6 for R, respectivly. The multiple of resistance reversal of B vs SA was 6.6. The relative reversal rate was 87.6%. CONCLUSION: A combination of B and SA may enhance the antimalarial effect against P. b and reverse the SA-resistance of P. b and delay the occurrence of resistance to SA in N. PMID- 12567721 TI - [Introduction to two simplified methods of COPT for diagnosing of schistosomiasis]. PMID- 12567720 TI - [Studies on the recurrent attacks of acute adenolymphangitis due to Malayan filariasis]. AB - OBJECTIVE: To explore the role of filarial and bacterial infections in the recurrent attacks of acute adenolymphangitis due to malayan fialriasis. METHODS: 1. To observe the seasonal fluctuation of acute attacks by performing monthly follow-up on patients with history of acute attacks in recent years. 2. To study the relationship between bacterial infection and filarial adenolymphangitis by performing bacteria culture and anti-streptolysin O test. 3. To investigate the variation of acute attacks by controlling filariasis transmission or by treating patients with a history of recurrent acute attacks. RESULTS: 1. The peak of acute attacks in patients coincided with the peak of vector transmission season. 2. Of the 97 cases examined by bacteria culture, 90 cases were negative; of the 255 cases examined by anti-streptolysin O test, the titres in 94.1% (143/152) of the cases with first attack and simple adenolymphangitis were within normal limits, however, the titres in 27.2% (28/103) of the cases complicated with elephantiasis were increased. 3. The acute attack rate of adenolymphangitis per year reduced significantly in cases with first attack and simple adenolymphangitis after effective control of filariasis transmission. 4. There was no evidence of the reduction of acute attacks by treating patients with DEC alone. CONCLUSION: In malayan filariasis endemic areas, the main causes of recurrent attacks of acute adenolymphangitis might be the repeated filarial infections due to the persistence of filariasis transmission. PMID- 12567722 TI - [Studies on the continuous culture and pathogenicity of Entamoeba gingivalis]. AB - OBJECTIVE: To establish a method for continuous culture of Entamoeba gingivalis (E. g.) and study the pathogenicity of E. g.. METHODS: The culture conditions of E. g. were compared by observing its growth and reproduction. The pathogenicity of E. g. was studied by investigating its infection rate in humans and observing the relationship between E. g. infection and periodontal abscess in rats and the in vitro effect of some drugs. RESULTS: The optimal culture condition of E. g. was modified LES medium containing Locke's solution with 20% bovine serum, penicillin, streptomycin and rice starch. Reproduction of E. g. peaked at the fourth day of incubatoin, E. g. could live continuously for 120 to 168 h. The infection rate of E. g. was higher in patients with periondontal diseases than in healthy groups. The animal test showed that E. g. could cause periondontal abscess in rats. The levels of acid phosphatase and MDA were higher in patients with E. g.-positive periondontal disease than in healthy individuals. Metronidazole and SBF are effective against E. g.. CONCLUSION: E. g. could grow very well in modified LES medium, and cause periondontal abscess in patients with low immunity. PMID- 12567723 TI - [Circulating antigen detection in schistosomiasis japonica patients by sandwich ELISA using polyclonal and monoclonal antibodies]. AB - OBJECTIVE: To detect circulating antigen in the patients with schistosomiasis. METHODS: Polyclonal antibody(PcAb)-monoclonal antibody(McAb) sandwich ELISA. RESULTS: Sera from 150 patients with schistosomiasis were detected, the average sensitivity was 84.7%(72.0%-91.0%). No false positive reaction was detected in 40 normal controls and none but 2 patients(2/10) with paragonimiasis showed cross reactions among 74 cases with other parasitic infections. CONCLUSION: PcAb-McAb sandwich ELISA is a relatively ideal method for circulatling antigen detection. PMID- 12567724 TI - A NMRI suckling mouse model for the evaluation of infectivity of Cryptosporidium parvum oocysts. AB - OBJECTIVE: To evaluate the infectivity of Cryptosporidium parvum oocysts in NMRI suckling mouse. METHODS: Four-day-old SPF NMRI suckling mice were inoculated with different amounts of oocysts by oral gavage. On day 7 after inoculation, suckling mice were sacrificed, and a suspension was prepared by homogenizing the intestinal tract from pylorus to anus. A mouse was considered infected when oocysts were found in smears of the intestinal content suspension stained with carbo lfuchsin solution. The infectivity of oocysts was evaluated as measured by the percentage of infected mice in each group. RESULTS: Mice receiving 1,500 or 2,000 oocysts were all infected. The percentages of infected mice were 88, 74, 51 and 28 respectively after ingestion of 1,000, 500, 250 and 100 oocysts. The percentage of infected mice was 9.5% after ingesting as few as 50 oocysts. CONCLUSION: This model is convenient for evaluation of the infectivity of C. parvum oocysts. PMID- 12567725 TI - [Correlation analysis of vaginal trichomoniasis and Lactobacillus]. PMID- 12567726 TI - Chemotherapeutic efficacy of praziquantel in rats with protective immunity to Clonorchis sinensis infection. AB - OBJECTIVE: To study the synergic effect of praziquantel (PZQ) and host acquired immunity on Clonorchis sinensis. METHODS: Acquired immunity to C. sinensis was induced by immunization with crude adult worm antigen (AW Ag) and excretory secretory antigen (ES Ag) or infection with C. sinensis metacercariae. The effect was assessed by the worm reduction rate compared with the control groups after challenge infection with 50 metacercariae and treated orally with a subcurative dose of praziquantel (50 mg/kg). Significant test was performed by analysis of variance (ANOVA) and Npar1 way Kruskal-Wallis test. All calculations were performed by PC-SAS system. RESULTS: 1. PZQ was more effective against C. sinensis larvae than against adult worms in the control (P < 0.001), ES Ag (P < 0.01) or crude AW Ag immunization group (P < 0.001). 2. As compared with the control, the worm reduction rate after challenge infection was significantly higher (P < 0.001) in ES Ag immunized group (35.6%) and metacercaria infection group (97.5%) and less in crude AW Ag group (23.4%). The PZQ efficacy was significantly enhanced in ES Ag immunized group. CONCLUSION: The efficacy of PZQ against C. sinensis could be synergically enhanced in rats by inducing host acquired immunity. PMID- 12567727 TI - [Changes in T-lymphocyte subsets of peripheral blood in patients with filarial chyluria]. AB - OBJECTIVE: To study the changes in T-lymphocyte subsets CD4+:CD8+ of peripheral blood in 29 patients with present chyluria(PPC), 29 patients with chyluria history but without chyluria (PNPC) and 38 healthy controls. METHODS: The determination of CD3+, CD4+ and CD8+ was conducted using test reagents kits. RESULTS: The percentage of CD3+, CD4+ cell were signfcantly decreased in PPS group than in PNPC and healthy control group, the ratio of CD4+/CD8+ being under 1.0. The T-lymphocyte subsets (CD3+, CD4+, CD8+) and CD4+/CD8+ ratio of both PNPC and healthy control group were all within normal range. CONCLUSION: The immune function of the patients with filarial chyluria was impaired in terms of the changes in T-lymphocyte subsets. PMID- 12567728 TI - [Observation on the effect of five years' prevention and treatment of schistosomiasis in Nanjian mountain area]. PMID- 12567729 TI - [Isoelectric focusing analysis of Echinococcus multilocularis protoscolex antigens]. AB - OBJECTIVE: To understand the components of the crude antigens of Echinococcus multilocularis protoscoleces. METHODS: Isoelectric focusing was carried out to fractionate the protein components of E. multilocularis protoscoleces. The individual fractions were collected and subjected to SDS-PAGE, the gels were analyzed by a gel analysis system. RESULTS: The pI values of individual fractions varied from 4.0 to 8.5, while the molecule weights of main protein bands were 72/73, 62/63, 42/44, 35/36, 34 and 21/22 kDa. CONCLUSION: An alternative separation method of crude antigens of Echinococcus multilocularis was established. PMID- 12567730 TI - [Experimental observation on anaphylaxis induced by Echinococcus infection in Meriones unguiculatus]. AB - OBJECTIVE: To observe the anaphylaxis induced by Echinococcus granulosus or E. multilocularis (E. g. and E. m.) infection in Meriones unguiculatus. METHODS: 56 and 48 gerbils were intraperitoneally inoculated with E. g. protoscoleces and E. m. tissue homogenate. After three months, the anaphylactic responses initiated by intraperitoneal challenge injection with E. g. cystic fluids or partially purified antigen or E. m. metacestode homogenate antigen were observed. IgE antibody and direct eosinophilic leukocyte count (DELC) were detected at pre postchallenge. RESULTS: The infection rates for E. g and E. m. in 56 and 48 gerbils were 89.3% and 97.9%, respectively. Among these groups, the lowest rate of anaphylaxis was 62.5%, the highest was 100%. The lowest rate of anaphylactic shock was 12.5%, and the highest 16.7%. After the gerbis were infected with E. g. and E. m., both IgE antibody and DELC increased gradually. At 60 min post challenge, IgE antibody level were significantly decreased, while DELC showed a sharp increase. CONCLUSION: Anaphylaxis could be induced in gerbils by Echinococcus infection followed by a challenge injection. Different antigens lead to different and degrees among the anaphylactic responses in the gerbils, however, there were no significant differences between E. g. and E. m. infections in gerbils. PMID- 12567731 TI - [The effect of artesunate in preventing the populations from Schistosoma japonicum infection during flood-control]. AB - OBJECTIVE: To evaluate the prophylactic effect of artesunate in high-risk populations who had contacted the infested water. METHODS: From the 7th day post exposure to infested water, a total of 17,031 people who had contacted the infested water for more than 20 days had been treated with artesunate at a dose of 300 mg once a week for three successive weeks. On day 17 after the last medication, they were examined for schistosomiasis and followed-up 60 days later. RESULTS: No acute schistosomiasis case was found. ELISA-positive was found in 204(1.20%) and COPT > 3% was found in 195 cases (1.14%). The side-effects were slight. CONCLUSION: Artesunate is highly effective and safe for the prevention of schistosomiasis. PMID- 12567732 TI - [Advances in the candidate vaccine antigens against Schistosoma japonicum]. PMID- 12567733 TI - [Progress in molecular biological research on Sarcocystis species and other cyst forming Coccida]. PMID- 12567734 TI - [Epidemiological survey of malaria in Yuanjiang County, Yunnan Province]. PMID- 12567735 TI - [Diagnosis of cryptosporidiosis by a rapid sandwich-ELISA]. PMID- 12567736 TI - [Diagnosis of Clonorchis sinensis infection by ultrasonography]. PMID- 12567737 TI - [28 cases of enterobiasis presenting as enuresis]. PMID- 12567738 TI - Identified circadian rhythm genes of ciliary epithelium with differential display. AB - PURPOSE: To identify differential genes expressed in the rabbit ciliary epithelium during the circadian cycle of aqueous flow. METHODS: Total RNA from ciliary epithelium of rabbits at 8 AM (light on 1 hour) and 8 PM (light off 1 hour) were compared by differential display reverse transcription-polymerase chain reaction (DD RT-PCR), using 6% denaturing polyacrylamide electrophoresis, choose differential display bands, cut and reamplify with the same primer, clone and sequence. Search the database of Genbank, prolong them with 5' RACE and 3' RACE technique then clone, sequence and search database of Genbank. RESULTS: 93 Significant differences gene expression were detected between light on and light off in the rabbit ciliary epithelium. CONCLUSION: Differential display is a powerful tool to screen differentially expressed genes in circadian rhythm of ciliary epithelium. PMID- 12567739 TI - The preliminary experimental study of induced differentiation of embryonic stem cells into corneal epithelial cells. AB - PURPOSE: To study preliminarily induced differentiation of embryonic stem cells into corneal epithelial cells in vitro. METHODS: Murine embryonic stem cells were co-cultured with Rabbit limbal corneal epithelial cells in Transwell system to induce differentiation. Mophological and immunohistochemical examination were implemented. RESULTS: The induced cells from embryonic stem cells have an epithelial appearance. The cells formed a network and were confluent into film gradually after being co-cultured with rabbit limbal corneal epithelial cells for 24-96 hours. The cells ranged mosaic structure and localized together with clear rim. Most of the cells showed polygonal appearance. Transmission electron microscope showed lots of microvilli on the surface of induced cells and tight junctions between them. These epithelial-like cells expressed the corneal epithelial cell specific marker cytokeratin3/cytokeratin12. CONCLUSION: The potential mechanism of the differentiation of murine embryonic stem cells into corneal epithelial cells induced by limbal corneal epithelial cell-derived inducing activity is to be further verified. PMID- 12567740 TI - Vitreous cavity-associated immune deviation induced by retinal S antigen. AB - PURPOSE: To determine whether the vitreous cavity (VC) supports the induction of deviant immune responses to retinal soluble(S) antigen and to observe the influence of interleukin-1 (IL-1) on the immunologic properties of the VC. METHODS: Retinal S antigen was inoculated into the anterior chamber(AC) and the VC in Wistar rats. Seven days after antigen inoculation, the recipient animals were immunized with S antigen and complete Freund's adjuvant. Delayed-type hypersensitivity (DTH) was assessed by footpad challenge. To alter systemic immune conditions, IL-1 was administrated by intraperitoneal injection. RESULTS: Antigen-specific DTH did not develop in rats in which S antigen was injected into the AC and the VC. By contrast, when IL-1 administrated systemically, S antigen was injected into the AC and VC elicited strong DTH. CONCLUSION: The VC supports immune deviation for soluble antigen by acitivity suppressing antigen-Specific DTH. Systemic administration of exogenous IL-1 eliminates the capacity of the VC to support immune deviation to soluble antigen locally injected. PMID- 12567741 TI - Effect of hypericin on confocal imaging of Ca2+ signaling in cultured human retinal pigment epithelium. AB - PURPOSE: To investigate the mechanism of the Ca2+ signaling in cultured human retinal pigment epithelial (RPE) cells with the protein kinase C(PKC) specific inhibitor-hypericin stimulation. METHODS: Cultured human RPE cells were analyzed using the fluorescence Ca2+ dye fluo-3 AM and laser scanning confocal microscope(LSCM) after stimulation with 100 nM phorbol 12-myristate 13 acetate(PMA) and (or) 5 concentrations of hypericin(1, 2, 3, 4 and 5 microM). RESULTS: The normal fluorescence in RPE cells was strong and distributed throughout the cells. The nucleus appeared to be more fluorescent than the cytoplasm. After stimulation with PMA alone or 5 concentrations of hypericin, a rapid decrease in flurescence intensity was observed. There was no obvious difference in decreased curve among 5 concentrations. However, after stimulation with a 24 hr preincubation of PMA and 5 concentrations of hypericin, a further decrease was not observed. CONCLUSION: Fluo-3 AM appears to be a good indicator of the change in Ca2+ occurring in RPE cells and hypericin is a strong inhibitor of Ca2+ influx channel. Hypericin has potential as a therapeutic drug for proliferative vitreoretinopathy(PVR), the inhibitory effect on PVR might be caused by blocking the PKC activity and inhibiting Ca2+ influx pathway. PMID- 12567742 TI - Construction of the enhanced yellow fluorescent protein expression vector carrying IFN-gamma gene. AB - PURPOSE: To construct the enhanced yellow fluorescent protein (EYFP) vector carrying interferon-gamma gene (ifn-gamma) in order to provide an ideal reporter in the expression of ifn-gamma and location of protein in vitro and in vivo. METHOD: According to the nucleotide sequence of ifn-gamma gene, a pair of oligonucleotides was designed as primer whose two end contained nucleotide sequence of EcoR V and Not I restriction endonuclease respectively. The gene encoding for inf-gamma was amplified using PCR technqiue. After the PCR product was retrieved and purified, it was digested with EcoR V and Not I restriction endonuclease, and then cloned into the plasmid pIRES-EYFP. The recombinant plasmid pIRES-EYFPIFN-gamma was identified by restriction endonuclease enzyme analysis and DNA sequence analysis. RESULTS: The ifn-gamma was successfully amplified and verified by partial DNA sequence analysis. The recombinant plasmid was correctly screened. CONCLUSION: The EYFP expression vector carrying ifn-gamma gene was successfully established. This research work has formed a base for monitoring the ifn-gamma gene expression and protein position in living cells. PMID- 12567743 TI - A study on the preservation of fresh amniotic membrane. AB - OBJECTIVE: To establish the standard preservation methods of fresh amniotic membrane for clinical use. METHODS: Human placentas were collected aseptically from selective caesarean sections in normal women in time. Amniotic or placental membrane were peeled and preserved in N. S, P. B. S or DMEM at 4 degrees C or cultured in DMEM at 37 degrees C, 5% CO2. Trypan-blue staining, light and electronic microscopy were observed every six hours after preservation. RESULTS: Seventy percent of amniotic epithelial cells survived after preservation in N.S for 6 hours, PBS 12 hours, DMEM 24 hours and 1 week in tissue culture. The amount of living epithelial cells maintained in placental membrane preservation was less than that in amniotic membrane preservation at the same time (t-test, P < 0.01). No collagen degeneration was found during preservation. CONCLUSION: Preservative solution and time will affect the maintenance time of fresh amniotic membrane greatly. Fresh amniotic membrane should be preserved within 6 hours in N. S, 12 hours in P. B. S, 24 hours in DMEM at 4 degrees C and 1 week in tissue culture for clinical use. PMID- 12567744 TI - A case of Erdheim-Chester disease with bilateral orbital involvement. AB - PURPOSE: To describe a case of Erdheim-Chester disease with bilateral orbital involvement. METHODS: A 43-year-old female with bilateral proptosis was presented. Its clinical features, image findings, pathological character and therapeutic effect were evaluated. RESULTS: CT demonstrated bilateral, diffuse orbital mass. Histopathologic assessment revealed a diffuse xanthogranulomatous process with clusters of lipidladen histocytes. Numerous Touton giant cells were scattered throughout the lesion. Renal and heart failure happened during a 6-year follow-up period. Long bones roentgenogram demonstrated diffuse symmetrical sclerosis with extensive, lytic lesions. Systemic administration of corticosteroids, chemotherapy, immunoglobulin and traditional Chinese medicine showed good therapeutic result. CONCLUSIONS: An administration of systemic corticosteroids, chemotherapy, immunoglobin and traditional Chinese medicine can control Erdheim-Chester disease. Further exploration of its pathogenesis and collection of useful clinical data are required. PMID- 12567745 TI - Pathological, ultrastructural and immunohistochemical observations of adenoma of retinal pigment epithelium. AB - PURPOSE: To Study the clinical, pathological, ultrastructural and immunohistchemical characters of adenoma of the retinal pigment epithelium in order to offer evidence to diagnose this tumor. METHODS: Routine paraffin slices HE stain, histochemistry PAS and VG stain, transmission electron microscopy, and immunohistochemistry for S-100 and vimentin with LSAB method were used. RESULTS: The tumor cells were oval and cuboidal in shape. Part of the tumor had a tubular arrangement. Around the sheets of tumors cells there was a large amount of uniform red stick-like substances. The above matter represented positive in PAS stain. Most of the above matter was yellow, while less of the matter showed red in VG stain. Transmission electron microscopy showed that there were tight junctions between tumor cells. Immunohistochemistry showed positive for S-100, negative for vimentin. CONCLUSIONS: The ultrastructural and immunohistochemical characters of the adenoma of retinal pigment epithelium are consistent with the retinal pigment epithelium. PMID- 12567746 TI - The analysis of electroretinography of diabetes mellitus. AB - PURPOSE: In order to get deeper understanding of Diabetic Retinopathy(DR), we analyzed and evauated the results of the amplitude and latency of F-ERG a-wave, b wave and the total amplitudes of oscillatory potentials(OPs). METHODS: F-ERG of 105 eyes from 55 cases of DM were diagnosed by the medical department from July 1997 to July 1998. The 105 eyes were examined by ophthalmoscope and fluorescing in angiography and divided into there groups: 22 eyes with DM without DR(NDR), 56 eyes with background DR(BDR) and 27 eyes with proliferate DR(PDR). In addition, 30 eyes were regard as normal control group(NCG). We used VATA-2000 type vision electrophysiological instrument and inter-national standard for clinical ERG to do measure meat and recording automatically by computer. RESULTS: 1. The proportion of eyes number of invisible wave of a-wave, b-wave of F-ERG and Ops increased with the development of DR. 2. There were significant differences(P < 0.01) in the latency of a-wave between NCG and BDR and statistical significance(P < 0.05) between BDR and NDR. There were significant differences(P < 0.01) in the amplitude of b-wave among NCG and BDR, NCG and PDR, NDR and BDR, NDR and PDR. 3. The total amplitudes of OPs lowered with progressions of DR. There were significant differences(P < 0.01) in tota amplitudes of OPs between NCG and NDR, NCG and BDR, NDR and BDR, statistical significance(P < 0.05) between NDR and PDR. 4. There was no correlation between each index and the duration of the disease (P < 0.05). CONCLUSION: The amplitude of a-wave, b-wave, and total amplitudes are the targets for early diagnosis of DR. The combined analyses of the three indexes of ERG can determine the severity, curative effect, and prognosis of the disease. PMID- 12567747 TI - Motion perception of short-wavelength sensitive cones in glaucoma using random dots moving. AB - PURPOSE: To determine whether motion perception of short-wavelength sensitive(SWS) cones is capable of predicting primary open angle glaucoma(POAG) optic neuropathy. METHODS: Motion perceptions of SWS cones were isolated by Blue random dot stimulus displayed on the Yellow background, then the detecting displacement threshold of motion perception from SWS cone in POAG was measured, and compared with the age-matched normal group. RESULTS: The detecting displacement thresholds of motion perception from SWS cones in 8 sites of 15 degrees were all elevated in POAG compared with the normal group. CONCLUSION: These findings suggest that motion perception of SWS cones may reveal preclinical visual nerve damage in early POAG. PMID- 12567748 TI - A study of factors related to the incidence of cataract in patients with non insulin dependent diabetes mellitus. AB - PURPOSE: To investigate the factors related to the development of cataract in patients with non-insulin dependent diabetes mellitus(NIDDM). METHODS: 792 NIDDM patients received ophthalmologic examinations including visual acuity, external status of the eyes, slit lamp microscopy and ophthalmoscopy. Glucose, urea nitrogen(BUN), creatinine(Cr), urine acid(UA), N-acetyl-beta 2-D-glucosaminidase (NAG), beta 2-microglobulin(beta 2-MG) and serum albumin in blood were quantitatively tested. Glucose, pH value, protein, cells, cast and ketobodies in urine were assayed. Diagnosis of cataract was based on lens opacities classification system II. Any patient meeting "NII", "CII" or "PII" level was diagnosed as cataract. RESULTS: The incidence of cataract in this group of NIDDM was 62.37% (494/792), which significantly related to the duration of the disease course, but not to the sex of the patient. The occurrence rate of cataract in patients suffering from NIDDM of less than five years duration, from five to ten years, and more than ten years was 49.67% (228/459), 71.84% (125/174), and 88.68% (141/159), respectively. The occurrence of cataract in patients diagnosed of the disease from five to ten years and more than ten years was much higher than that of those with the course of the disease less than five years(P < 0.05 and P < 0.001, respectively). Rising concentrations of blood urea nitrogen, creatinine, glycosylated hemoglobin HbA1c(G-HbA1c), N-acetyl-beta 2-D-glucosaminidase(NAG) and beta 2-microglobulin(beta 2-MG) indicated malfunction of the kidneys, and the rate of cataract occurrence in these patients was higher. CONCLUSION: This study indicates that prolongation of the duration of non-insulin dependent diabetes mellitus, renal dysfunction, as well as poor blood glucose control, may accelerate the development of cataract. PMID- 12567749 TI - Treatment of neovascular glaucoma using trabeculectomy combined with implantation of silicon rubber slice. AB - PURPOSE: To observe clinical effect of treatment for neovascular glaucoma using trabeculectomy combined with implantation of silicon rubber slice. METHODS: 28 cases(28 eyes) with neovascular glaucoma were performed trabeculectomy combined with a piece of 3 x 5 mm silicon rubber implantation, and their lowing intraocular pressure effect was observed. RESULTS: All patients were follow-up for 9-24 months (mean: 18.3 months). The average intraocular pressure at last visit was 21.65 mmHg (rang: 17.30-28.97 mmHg, 1 mmHg = 0.133 kPa). The symptom of eye pain was disappeared after surgery. CONCLUSION: Trabeculectomy with a piece of silicon rubber implantation is ideal therapy for neovascular glaucoma. PMID- 12567750 TI - The clinical study of Healon 5 in phacoemulsification and IOL implantation. AB - PURPOSE: To compare a new ophthalmic viscoelastic device (OVD) Healon 5 with two other kinds of OVDs commonly used in China during phacoemulsification and intraocular lens (IOL) implantation in terms of removal time, intraocular pressure, corneal and anterior chamber response. METHODS: This prospective randomized study, in which patients and observers are masked, comprises niety eyes. They were randomly divided into 3 groups with different OVDs. Healon 5 (sodium hyaluronate 5,000 2.3%) was compared with Healon GV (sodium hyaluronate 7,000 1.4%) and Iviz (sodium hyaluronate 1.0%). The surgeries were performed with temporal corneal incision, phacoemulsification in situ and foldable lens intracapsular implantation. The characteristic and the removal time of the viscoelastic material as well as the postoperative IOP, corneal reaction and anterior chamber reactions of the eyes were observed preoperatively and 24 hrs postoperatively. Exclusion criteria were glaucoma, a preoperative dilated pupil diameter smaller than 5.0 mm, proliferative diabetic retinopathy, significant corneal pathology or a history of uveitis. RESULTS: The removal time was 47.42 +/ 13.09 seconds in Healon 5, 16.50 +/- 5.45 in Healon GV and 15.83 +/- 7.40 in Iviz. The removal time of Healon 5 group was significantly longer than that of the other groups (P = 0.000). There is no significant IOP change 24 hours postoperatively and the difference of intraocular pressure among three groups was not significant (P > 0.05). Healon 5 group had the slightest corneal and anterior chamber response, followed by Healon GV and Iviz. CONCLUSION: Although takes longer time to remove, Healon 5 is an ideal viscoelastic material for its superior space maintenance capacity and high dispersity which provides superior protection of endothelial cells compared to Healon GV and Iviz. PMID- 12567751 TI - [Immunohistochemistry research on keratoconus]. AB - PURPOSE: Keratoconus is a noninflammatory disease characterized by thining and ectasia of the central cornea. The etiology and pathogenesis of keratoconus remain unclear. We studied the composition of basement membranes (BM) in normal, scarred and keratconus corneas to find out possible changes specific for keratoconus. METHODS: Paraffin-embeded sections of normal, scarred and keratoconus corneas were immunostained with antibodys against type IV collagen, Fibronectin (Fn) and Laminin (Ln). RESULTS: Keratoconus had increased staining of the epithelial BM for alpha 1-alpha 2 chain of type IV collagen and Fn, especially in scarred corneal regions. A similar enhancement was also observed in scarred corneas, and there was no significant difference between keratoconus and scarred cornea. CONCLUSION: A process similar to wound healing would largely contribute to the differences seen in the keratoconus. PMID- 12567752 TI - [Comparing cryopreserved with fresh corneas on clinical application in penetrating keratoplasty]. AB - PURPOSE: To evaluate the role of cryopreserved corneas on penetrating keratoplasty (PKP) by comparing them with fresh corneas on clinical application. METHODS: Third-six excellent condition patients were chosen and randomly divided into two groups. They were performed PKP by using fresh and cryopreserved corneas respectively. The graft transparency, epithelial healing speed of graft, tear membrane, graft thickness, endothelium density and visual acurity were emphasized respectively. RESULTS: When fresh and cryopreserved corneas were used on PKP as the donor respectively, postoperative time to recover normal break-up time (BUT) was two to six months average (4.7 +/- 0.3) months and two to eight months, average (5.6 +/- 0.4) months and time to recover normal Schirmer's test was one to four months average (2.1 +/- 0.3) months and 1-3.5 months, average (1.7 +/- 0.5) months. The graft's thickness, stabilized visual acurity and graft's rejective rate were 0.56-0.68 mm (average 0.62 mm) vs 0.54-0.62 mm (average 0.59 mm), (0.46 +/- 0.03) vs (0.44 +/- 0.05) and 20% vs 19% respectively when fresh vs cryoperserved corneas. No statistical significance between fresh and cryopreserved corneas (P > 0.05). But the grafts behaved intact epithelium and continuing transparency when fresh corneas as donors, while the graft's epithelial defects healed after three to five days of the operation and edematous grafts were reclear after two to three weeks of the operation when cryopreserved corneas as donors. Graft's endothelium density in fresh corneas group 2,043-2,210 cells/mm2 (average 2,135 cells/mm2) was high than in cryopreserved corneas 1,240 1,860 cells/mm2 (average 1,672 cells/mm2) (P < 0.01). CONCLUSION: There were no statistical significance on the transparent rate of grafts, grafts thickness, visual acurity and graft's rejective rate between fresh and cryopreserved corneas. But graft's epithelium healing was delayed and endothelium density was lower when cryopreserved corneas as donors. PMID- 12567753 TI - [The application of hansatome microkeratome in LASIK]. AB - OBJECTIVE: To evaluate the application of Hansatome in excimer laser in situ keratomileusis(LASIK). METHODS: Two hundred and sixty-six cases(514 eyes) with myopia were treated with the Hansatome microkeratome and the excimer laser. According to the preoperative spherical equivalent refraction, the patients were divided into three groups: < -8.0 D, -8.0 D(-)-15.0 D and > -15.0 D. The advanced rails of Hansatome microkeratome was revolving, then the stalk of corneal cap was over cornea. Postoperatively for six months all patients had been followed up on visual acuity, refractive status and complication. RESULTS: The uncorrected visual acuity in three groups was 0.5 or better in 303, 144 and 28 of the eyes, 278, 124 and 10 had vision of 1.0 or better six months after operation. The refractive status tended to be stable about four to six months after the operation. The complication of operation included corneal epithelial excoriation, incomplete cap, free cap, interfacial foreign bodies, glare, etc. The learning curve of the technique requested half a year. CONCLUSION: The Hansatome microkeratome in LASIK is effective for the correction of myopia and it has better safety. The complication of the stalk of corneal capover cornea might be less than which of the inside the nose. PMID- 12567754 TI - [Removal of congenital persistent pupilary membrane with sutureless clear corneal small incision]. AB - PURPOSE: To investigate a new way in the treatment of serious congenital persistent pupilary membrane. METHODS: In four cases of six eyes, after temporal clear corneal tunnel incision was made, viscoelastic material was injected into the anterior chamber, and the persistent pupilary membrane was cut down at the very point connected to the iris. RESULTS: There is no ocular hypertention and hyphema after operation. After one year's follow-up, the lens remained transparent in 3 cases(4 eyes). There was corneal astigmatism of 0.12 to 0.25 D after operation. And only a few corneal endothelial cells were lost. CONCLUSION: There are few postoperative complications and corneal astigmatism to incise congenital persistent pupilary membrane by temporal clear corneal tunnel incision. It is an ideal way to treat serious persistent pupilary membrane. PMID- 12567755 TI - [Character of alpha-crystallin molecular chaperone activity in selenite cataract]. AB - PURPOSE: To evaluate the character of alpha-crystallin molecular chaperone activity in selenite cataract. METHODS: alpha-Crystallin from lenses of normal young rats and lenses with selenite cataract were separated and purified by gel filtration chromatography. The chaperone activity of alpha-crystallin was assayed by mearsuring heat-induced aggregation and scattering of catalase. RESULTS: alpha Crystallin in both normal and cataractous lenses specifically prevented catalase against thermal aggregation compared with control proteins. The preventive ability in alpha H-crystallin was less than in alpha L-crystallin. Chaperone activity was diminished in selenite cataract. There was statistically significant difference in chaperone activity between normal and cataractous lenses in both alpha H-crystallin and alpha L-crystallin respectively. CONCLUSION: The reduction of alpha-crystallin chaperone activity in selenite cataract by preventing thermal aggregation of catalase is an important process in cataract formation. PMID- 12567756 TI - [The cause and management of acute high intraocular pressure in phacoemulsification]. AB - OBJECTIVE: To investigate the cause and management of acute high intraocular pressure and shallow anterior chamber in phacoemulsification. METHODS: A retrospective study was done on 1,500 eyes performed phacoemulsification. Eleven of 1,500 eyes showed higher intraocular pressure and shallowing of the anterior chamber during phacoemulsification. But phacoemulsification was successfully achieved after giving hyperosmotic agents, increasing infusion pressure and reducing aspiration rate as well as flow rate. RESULTS: Of 11 eyes, 10 eyes were treated by management of non vitrectomy. One of 11 eyes was unresponsive to medical management and require the use of pars plana vitrectomy. Phaco was performed in all patients. In 4 of 11 eyes, visual acuity postoperatively were 0.7 or better. Five of 11 eyes were 0.3 to 0.5. One eye was 0.05 and one was HM/30 cm. CONCLUSION: Aqueous misdirection, retrobulbar hemorrhage and intraoperative suprachoroidal hemorrhage are common causes of acute high intracular pressure in phaco. Most of patients could be performed phaco after medical treatment. If patients failed to improve after medical treatment, par plana vitrectomy is a useful method. PMID- 12567757 TI - [Phacoemulsification in patients with cataract and overfiltering bleb after antiglaucoma surgery]. AB - PURPOSE: To evaluate the effect of phacoemulsification in patients with cataract, overfiltering bleb, and low-tension after antiglaucoma surgery. METHODS: Phacoemulsification and implantation of a foldable intraocular len was performed in six patients(7 eyes) with overfiltering bleb, low-tension, and cataract. The complications, blebs, intraocular pressures(IOP), and visual acuity were observed postoperatively. RESULTS: No severe complication was found. The overfiltering blebs constricted and their walls became thicker three months after the operation. The visual acuity and IOP were improved (P < 0.05). CONCLUSION: Phacoemulsification may be another way to treat the patients with cataract and overfiltering bleb, low-tension after antiglaucoma surgery. PMID- 12567758 TI - [The inhibitory effect of mitomycin C applied by intracapsular method on rabbit lens epithelial cells: an experimental study of after-cataract]. AB - OBJECTIVE: To study the inhibitory effect of Mitomycin C applied by intracapsular infusion method on rabbit lens epithelial cells and its intraocular toxicity. METHODS: 0.2 ml Mitomycin C with different concentration (0.1 mg/ml, 0.2 mg/ml, 0.4 mg/ml) was injected into the capsular bags of the rabbits' lens during the hydrodissection of phacoemulsification. The drug was exposed to the lens epithelial cells directly and transiently. In the two-month follow-up after the operation, the posterior capsule opacification and the change of intraocular pressure were observed and compared between the control group and the experimental group; the histopathologic and the ultrastructural changes were also observed. RESULTS: For clinical study in the rabbit, the posterior capsule opacification only appeared in the control group two weeks after the operation. Four weeks after the operation, the posterior capsular opacification in the eyes of experimental group was not so obvious as that of the control group. The higher the concentration of the drugs, the less the posterior capsule opacification was shown. There was no obvious difference in the postoperative inflammation between the control group and the low, middle-concentration experimental group. Slight toxic reaction was shown on the rabbits' eyes of the high-concentration experimental group in the early stage after the operation. The histopathologic examination showed that the proliferation of lens epithelial cells in the control group was more obvious than that in the experimental group. Mitomycin C could cause the degeneration of lens epithelial cells. The degree of degeneration was related to the drug's concentration. The ciliary bodies of the rabbits' eyes in the high-concentration experimental group were infiltrated by inflammatory cells and bled slightly. CONCLUSION: Mitomycin C can inhibit the proliferation of lens epithelial cells effectively. Its intensity is related to the drug's concentration. It's a safe and effective drug in the prevention of after-cataract when the concentration of 0.1-0.2 mg/ml was applied in the capsular bag. PMID- 12567759 TI - [Hard nucleus chopping technique for non-phacoemulsification in small-incision cataract surgery: two-knife chopping]. AB - PURPOSE: To search for a practical nuclear chopping technique for non phacoemulsification in small-incision cataract surgery. METHODS: We designed two choppers for dividing nucleus in cataract surgery. We used them in our clinical practice of hard nuclear cataract surgery and thus improved the choppers constantly. RESULTS: Two hundred and forty-six cases (246 eyes) of cataract with their nuclei being Grade III or harder were involved in our observation. Nucleus had been divide into two halves at one chopping and the nuclear halves were delivered through a 4.5 mm incision safely in all cases. Visual acuity was 0.5 or better in 189 eyes (73.8%), 232 eyes (94.9%) and 241 eyes (97.9%) one day, seven days and three months respectively after surgery. Complications included corneal edema near the incision in 12% eyes and folding of the Descemet membrane in 25% eyes. CONCLUSION: Two-knife chopping technique has the advantages of reasonable design, small incision, little injury, no limitation by hard nucleus and easy manipulation. It is a practical and ideal chopping technique in non phacoemulsification small-incision cataract surgery. PMID- 12567760 TI - [Application of corepraxia in the implantation of intraocular lens]. AB - PURPOSE: In the cases of aphakia or cataract which complicated with iridosteresis, corectasis and pupil displacement, corepraxia was performed when implanting intraocular lens (IOL) in order to recover the normal shape and position of pupil, improve postoperative visual acuity and prevent monodiplopia and glare. METHODS: In the cases which have iridosteresis at the direction of 9-3 oclock or updrawn pupil, iris suture was performed at the incision directly(open method); while those at the direction of 3-9 o'clock, operation was performed outside the anterior chamber (close method). RESULTS: In 22 cases, the percentage which having visual acuity better than 0.5 is 65%, having nearly round shape pupil 67%, having pupil diameter less than 3 mm 73%, having normal pupil position 67%. CONCLUSION: All kinds of deformed pupil can be recovered to their normal shape, size and position by performing corepraxia while IOL was implanted. The prolene has light reaction and better effect in iris suture. PMID- 12567761 TI - [A model of calibrated optic nerve crush injury in rats]. AB - OBJECTIVE: To establish a calibrated optic nerve crush injury model in rats. METHODS: Twenty-eight healthy Sprague-Dawley (SD) rats were randomly divided into two groups. Seven rats were in normal control group, i.e. group A. In group B, model of partial optic nerve crush injury group, partial optic nerve crush injury model was induced in the right eyes of another 21 rats by a special designed optic nerve clip with 40-gram holding force at optic nerve 2 mm behind the eyeball clipped for four seconds to partially block the optic nerve axoplasmic transport. The left eyes served as a control. According to the sacrificing date of the rats, three subgroups in group B were further divided. Therefore, seven rats were in each subgroup. One, eleven and eighteen days later, 3% fast blue was injected into both superior colliculi to each subgroup and group A. The eyes were enucleated another three days later, flat mounts of the retinal from both eyes were prepared on a slide and observed under a fluorescence microscope. Four photos with 400 x magnification were taken from quadrants of the retina 1 mm away from the optic disc. The labeled retinal ganglion cells (RGCs) photos were counted by a computerized image analyzer. The labeled RGCs rate, a comparison of the labeled RGCs from both eyes in a rat were used for statistical analyzing. (The labeled RGCs rate = RGCs from injured right eye/RGCs from uninjured left eye x 100%). RESULTS: In group A, the group of normal control group, the labeling rate from the ratio of RGCs counting of right eye to left eye was 99.79% +/- 13.05% and from the ratio of the RGCs of left eye to right eye was 101.86% +/- 13.91%. In group B, model of partial optic nerve crush injury group, the labeled RGCs rate were 77.79% +/- 7.11%, 63.76% +/- 3.79% and 54.66% +/- 4.75% in day 4, day 14 and 21 respectively. CONCLUSION: Using a special designed clip with 40 gram holding force constantly pressed for four seconds to create an optic nerve crush injury model in rats. As time past along, it is revealed that the labeled RGCs rate gradually reduced. PMID- 12567762 TI - [Influence of cell cycle in human trabecular cells induced by dexamethsone]. AB - PURPOSE: To investigate the influence of dexamethesone (Dex) on cell cycle in human trabecular cells(HTC). METHODS: The trabecular specimens from human eyes were cultured in vitro. Then the cultured cells were immersed in DMEM solution containing 10(-7) M dexamethesone for three, five and seven days. The cell cycle was detected by Flow Cycometry. RESULTS: The G2 phase of cell cycle in HTC increased when HTC were treated by Dex. The G2 phase was 2% in control group. For three, five and seven days treated by Dex, the G2 phase were 8%, 16.3 and 19.4%. CONCLUSION: The influence of cell cycle might be induced by Dex, it may be related with the pathogenesis of glucocorticoid induced glaucoma. PMID- 12567763 TI - [The expression of growth factors in filtering area following sclerectomy and the interaction between growth factors and interferon]. AB - OBJECTIVE: To study the expression of beta-Fibroblastic Growth Factor (bFGF) and Transforming Growth Factor beta 1 (TGF beta 1) in filtering area following sclerectomy and the interaction between growth factors and interferon alpha-2b (IFN alpha-2b). METHODS: Immunohistochemical technique were applied to test the expression bFGF and TGF beta 1 in frozen sections of filtration area following sclerectomy on postoperative day three, five, seven and 14 in white rabbits, respectively. 16 eyes of eight rabbits in all were randomly and equally divided into two groups. The experimental eyes had subconjunctivally been administered IFN alpha-2b 5 x 10(5) IU 0.2 ml each time at filtering bleb following operation whereas the control eyes hadn't taken. RESULTS: In control group, positive cell and substance expressing bFGF and TGF beta 1 on postoperative day three were revealed in the filtration area, on postoperative day five and seven, they were gradually increased. On the postoperative day 14, all significant cells and substance markly decreased. In experimental group only small amount of positive cell and bFGF or TGF beta 1-expressing substance were sparely distributed in the filtration area. CONCLUSION: The filtration area after glaucoma filtering surgery was "bathed" in growth factors such as bFGF and TGF beta 1 which well-known play an important role in stimulating wound healing response, IFN alpha-2b had an inhibitory effect on bFGF or TGF beta 1 expression in wound environment. PMID- 12567764 TI - [Study on the dynamic changes of retinoblastoma gene of SO-Rb50 cell line]. AB - PURPOSE: To study the dynamic changes of Retinoblastoma Gene of SO-Rb50 cell line. METHODS: 1) Southern blot hybridization was used to analyse the tumor cell DNA of 327th passage of SO-Rb50 cell line. 2) A promoter and 27 exons of Rb gene were screened exon-by-exon by using PCR-SSCP at 415th and 713th passages of SO Rb50 cell line. 3) Three cell cloning strains named as MC2, MC3 and MC4 were isolated by single cell cloning technique from the SO-Rb50-775, and mutation of Rb gene were also screened exon-by-exon by using PCR-SSCP-HA in MC2-11, MC3-11, MC4-11 and MC3-138. RESULTS: The 3.5 Kb, 2.9 Kb and 1.0 Kb bands were deleted in the DNA of SO-Rb50-327 tumor cells, showing the deletion of Rb gene in SO-Rb50 cell line. Exon23 of 451th passage cells decreased one band; but exon 25 of 713th passage cells decreased two bands, indicating that exon 25 had new mutation. PCR SSCP-HA analysis of exon24 showed that MC4-11 and MC3-138 had abnormal bands, but MC2-11 and MC3-11 weren't found mutation of Rb gene. This result suggested that new mutation occurred to exon24 of MC3 during a long-term culture in vitro. CONCLUSION: Retinoblastoma gene mutation of SO-Rb50 cell line had dynamic changes during a long-term culture. PMID- 12567765 TI - [Correlations of Ki-67 expression and histopathologic characteristics in uveal melanomas]. AB - PURPOSE: To investigate the relations between the cellular proliferation and histopathologic variables in paraffin embedded uveal melanomas with the using of the monoclonal antibody Ki-67 as a proliferation marker. METHODS: Fifty-seven enucleated eyes were included for histopathological study and Ki-67 immunostaining. All these eye globes were enucleated between 1988 and 1997 without prior treatment. RESULTS: The score of Ki-67 index ranged from zero to 4.89%, mean (0.75 +/- 1.02) %. The mean score of Ki-67 was significantly higher in large tumors (P = 0.007) or in tumors that contained epithelioid cells (P = 0.037) when compared to medium size tumors or to spindle cell type tumors. There was a significant difference of the distribution of epithelioid cell and spindle cell when compared histologic cell type with tumor size (chi 2 = 4.528, P < 0.05). Higher proportions of large tumors were found in epithelioid cell group. No significant correlation was found between the Ki-67 index and tumor location, scleral extension and age. CONCLUSION: The Ki-67 index is significantly correlated with the histologic cell type and tumor size. PMID- 12567766 TI - [The study on effects of rhEGF on proliferation of human conjunctival epithelial cells]. AB - PURPOSE: To investigate the effects of Recombinant Human Epidermal Growth Factor (rhEGF) on proliferation of human conjunctival epithelial cells and optimal dose in vitro. METHODS: Cultured human conjunctival epithelial cells were plated in 96 well plates at a density of 2,000 cells per chamber, treated with different concentrations of rhEGF (100 micrograms/ml, 50 micrograms/ml, 20 micrograms/ml, 10 micrograms/ml, 5 micrograms/ml and 1 microgram/ml). After 24, 48, 72 and 96 hours of treatment, cell viability was assessed with MTT assay. RESULTS: There was no significant difference in all experimental groups and the control group in the first 24 horus. Experimental groups at a concentration of 50 micrograms/ml, 20 micrograms/ml and 10 micrograms/ml had significantly higher cell viability than which of the control group 48 hours later. All groups of rhEGF had significantly higher cell viability than which of the control group after 72 hours treatment, except for 100 micrograms/ml of rhEGF. Each group of rhEGF after 96 hours treatment had higher cell viability than which of the control group, and 100 micrograms/ml of rhEGF had lower cell viability than which of the other experimental groups, with significant difference. CONCLUSION: rhEGF enhance proliferation of human conjunctival epithelial cells effectively in vitro. The optimal concentration ranged from 10 micrograms/ml to 50 micrograms/ml. PMID- 12567767 TI - [Inhibition of cultured human retinal pigment epithelial cell by tissue-type plasminogen activator]. AB - PURPOSE: To study the inhibition effects of human retinal pigment epithelial (RPE) cells by tissue-type plasminogen activator(tPA). METHODS: The effects of tPA on human RPE cells proliferation were studied by viable cell counting and methyl thiazolyl tetrazolium (MTT) colorimetric assay. Flow cytometry (FCM) analysis was used to examine RPE cell cycles. RESULTS: The proliferation of human RPE cells could be inhibited by 0.1-3 micrograms/ml tPA (P < 0.05). The effects of inhibition were depended on the drug density and the action time directly (P < 0.01). 5 micrograms/ml tPA was deadly-toxic to human RPE cells. FCM revealed that the cells in S phase increased 9.8% (P < 0.05) and cells in G2M phase decreased 6.6% (P < 0.05), compared to controls. CONCLUSION: tPA at a certain dose range and period could inhibit human RPE cells proliferation without deadly-toxicity. The human RPE cells growth could be interfered in S phase by tPA. PMID- 12567768 TI - [A case report of idiopathic polypoidal choroidal vasculopathy]. AB - PURPOSE: To report a case of idiopathic polypoidal choroidal vasculopathy (IPCV) and its angiographic characteristics. METHODS: General ophthalmic check up, analyses of fundus picture, funds fluorescein angiography (FFA), indocyanine green angiography (ICGA) as well as literature review. RESULTS: The lack of typical choroidal neovascularization (CNV) on FFA and characteristic polypoidal choroidal vasculopathy on ICGA as well as its unsymmetrical manifestation and discrepancy with age-related macular degeneratio (AMD) on the fundus picture are typical signs for the diagnosis of IPCV. CONCLUSION: The angiographic features on FFA and ICGA are specific for the diagnosis of IPCV. Differentiating IPCV and AMD has clinical significance in prognosing the visual acuity, progress and management of the diseases. PMID- 12567769 TI - [Surgery treatment of retinal detachment with Marfan syndrome]. AB - PURPOSE: To detect the reattachment rate and visual function outcome of Pars Plana Lensectomy, vitrectomy and scleral encircling (PPL + PPV + SE) to treat retinal detachment (RD) with Marfan syndrome. METHODS: A retrospective study of 11 cases of 14 eyes with Marfan syndrome who submitted to PPL + PPV + SE. RESULTS: All the cases were followed up for four - 46 months (means 31.2 months). Retinal reattachment rate was 85.7%, visual acuity were 0.3 or better in eight cases. CONCLUSION: PPL + PPV + SE is a safe and effective treatment for retinal detachment with Marfan syndrome. PMID- 12567770 TI - [Antagonizing effects of novel multipeptid analogues on endothelin receptors and their pharmacological characteristics in cardiovascular system]. AB - AIM: To investigate the antagonistic effects of the novel compounds on vasoconstriction induced by ET-1 and the effect on the blood pressure of stroke prone spontaneously hypertensive rats. METHODS: Organ bath experiment and whole cardiac function experiment were used. RESULTS: The analogues of o-CPhe-D-Trp-D Phe(-X)-OH showed good ability against endothelin biological effects. When X was displaced by 3-F, 3-Cl or 4-Cl, the novel compounds inhibit the vascular constriction induced by ET-1 in a concentration-dependent manner, the IC50 +/- L95 were (0.09 +/- 0.05), (0.15 +/- 0.06) or (0.11 +/- 0.03) mumol.L-1 respectively. The blood pressure of stroke-prone spontaneously hypertensive rats was decreased. No significant effect on cardiac function of rats was discovered. CONCLUSION: The results demonstrate that among the six kinds of compounds, those with o-CPhe-D-Trp-D-Phe (-X)-OH configuration showed good biological effects. PMID- 12567771 TI - [Effects of chronic exposure to beta-amyloid-peptide25-35 on the mRNA expressions of voltage-gated outward potassium channel subunits in cultured rat hippocampal neurons]. AB - AIM: To investigate mRNA expression changes of voltage-gated outward potassium channel subtypes in cultured rat hippocampal neurons after chronic exposure to beta-amyloid-petitde25-35 (beta-AP25-35). METHODS: mRNA expression was detected by RT-PCR, comparative expression levels were determined by imaging densitometer. RESULTS: Delayed rectifying (Kv2.1, Kv1.5), transient outward (Kv1.4, Kv4.2) and large conductance calcium-activated (rSlo) potassium channel mRNA were expressed in cultured rat hippocampal. In the presence of beta-AP25-35 3 mumol.L-1 for 24 h, the relative expression level of Kv2.1 was significantly increased (n = 3, P < 0.05); the other subtypes were not changed obviously (n = 3, P > 0.05). The increase of Kv2.1 mRNA mainly happened between 24 and 36 h after exposure to beta AP25-35. After exposure to beta-AP25-35 for 60 h, Kv2.1 mRNA decreased significantly (n = 3, P < 0.01). CONCLUSION: The upregulation of Kv2.1 on transcription levels may be involved in the enhancement of delayed rectifying outward potassium (Ik) current induced by beta-AP25-35. PMID- 12567772 TI - Effects of benzyltetrahydropalmatine on the rapidly activating component of delayed rectifier potassium current in guinea pig ventricular myocytes. AB - AIM: To investigate the effect of benzyltetrahydropalmatine (BTHP) on the rapidly activating component of delayed rectifier K+ current (Ikr) in single guinea pig ventricular myocytes. METHODS: Whole-cell patch clamp technique was used to record Ikr. RESULTS: Ikr was blocked by 1-100 mumol.L-1 BTHP in concentration-, voltage-, and specifically frequency-dependent fashion, with IC50 of 13.5 mumol.L 1 (95% confidence range: 11.2-15.8 mumol.L-1). 30 mumol.L-1 BTHP reduced Ikr and Ikr.tail by (31 +/- 4)% and (36 +/- 5)% (n = 6, P < 0.01), respectively. The time constant for deactivation (tau') of the tail current was decreased by 30 mumol.L 1 BTHP from (238 +/- 16) ms to (196 +/- 14) ms, while drug had no any effect on the time constant for activation (tau) of Ikr,tail. CONCLUSION: BTHP inhibited Ikr in a frequency-dependent fashion. PMID- 12567773 TI - [Sex-difference on flutamide metabolism in rat liver microsomal cytochrome P450 1A2]. AB - AIM: To assess the sex-difference on flutamide metabolism in rat liver microsomes useing rat cytochrome P450, 1A2, inhibitory monoclonal antibody. METHODS: Liver microsomes were prepared from male or female rats. Protein concentration and total cytochrome P450 content were determined. Incubation mixture included liver microsomes (1.0 nmol.L-1), reduced form of nicotinamide adenine dinucleotide phosphate (NADPH, 0.1 nmol.L-1), CYP1A2 (1:400) and flutamide (2 mg.L-1). The incubation time was 30 min. The concentration of flutamide and its major metabolite 2-hydroxyflutamide were analyzed by reverse high-performance liquid chromatography. The mobile phase was a mixture of methanol-acetonitrile-water diethylether (40:20:35:1) with methyltestosterone as internal standard. The detection wavelength was 234 nm. The reaction mixture was extracted with acetic ether 4 mL. Sex-difference on flutamide metabolism was expressed as the difference between the concentration ratio of 2-hydroxyflutamide to flutamide in male and female rat liver microsomes. RESULTS: The recoveries of flutamide and 2 hydroxyflutamide for the proposed method were more than 75%. The formation of 2 hydroxyflutamide from flutamide was inhibited by CYP1A2 antibodies (1:400) in male and female rat liver microsome for 30 min of incubation time, but the inhibition of flutamide metabolism in female rat was stronger than that in male. The concentration ratios of 2-hydroxyflutamide to flutamide were (1.5 +/- 0.6) and (0.9 +/- 0.4) in male and female rat liver microsomes, respectively (P < 0.01). CONCLUSION: The results indicate that the activity of male rat CYP1A2 is higher than that of the female rat. There is difference in sex-related rate of flutamide metabolism in rat liver microsomes. PMID- 12567775 TI - [Effect of polysaccharide from Spirulina platensis on hematopoietic cells proliferation, apoptosis and Bcl-2 expression in mice bearing tumor treated with chemotherapy]. AB - AIM: To evaluate the effect of polysaccharide from Spirulina platensis (PSP) on hematopoietic cell proliferation, apoptosis and Bcl-2 expression in mice bearing tumor treated with chemotherapy. METHODS: The model of chemotherapy for transplant solid tumor in mice was established. The hematopoietic cell proliferation, apoptosis, Bcl2 expression and related cytokines were assayed by the technique of culture of hematopoietic progenitor cell, fluoromicroscope and light microscope, immunohistochemical method, and double antibody sandwich ELISA. RESULTS: PSP significantly ameliorated CFU-GM proliferation inhibition and hematopietic cells apoptosis induced by CTX. Moreover, PSP evidently increased the content of IL-1, IL-3, GM-CSF and TNF-alpha in serum and Bcl-2 expression of hematopoietic cells. CONCLUSION: PSP indirectly upregulated Bcl-2 expression of hematopoietic cells by promoting endogenous cytokines secretion which may be one of the mechanisms, by which PSP enhanced hematopoietic cell proliferation and inhibited its apoptosis in mice bearing tumor treated with chemotherapy. PMID- 12567774 TI - [mRNA expression of muscarinic receptors in spinal cord and brainstem in morphine dependent rats]. AB - AIM: To observe mRNA expression of muscarinic acetylcholine receptors in spinal cord and brainstem in morphine dependent or withdrawal rats. METHODS: The mRNA expression level of m1, m2, m3, m4 and m5 were determined by RT-PCR, the beta actin mRNA expression was used as internal control. RESULTS: The mRNA level of m1, m2, m3, m4 and m5 in spinal cord and m1 and m2 in brainstem were increased significantly during morphine dependence, and the levels of m1, m2, m3 and m4 in spinal cord and m1 in brainstem were decreased 1 h after the injection of naloxone (4 mg.kg-1, i.p.) in morphine dependent rats. Either scopolamine (0.5 mg.kg-1) or pirenzepine (10 mg.kg-1) was shown to significantly decrease the morphine withdrawal symptoms in rats. The levels of m1, m2, m3 and m5 in spinal cord were increased by pretreatment with pirenzepine and the levels of m2, m3 and m4 in spinal cord were increased by pretreatment with scopolamine. CONCLUSION: The adaptive expression of muscarinic receptors at spinal and supraspinal levels play important role in mediating morphine dependence and withdrawal in rats. PMID- 12567776 TI - [Synthesis and vasorelaxant activities of benzopyran-4-one hydrazone derivatives]. AB - AIM: In search of more potent, less toxic and selective potassium channel openers. METHODS: According to the structure-activity relationships of benzopyran compounds and the features of structures of aprikalim, dofetilide and nifekalant, twenty benzopyran-4-one hydrazone derivatives have been designed and synthesized from 4-cyanophenos through acetylation, Fries rearrangment, cyclization, hydrazone, substitution reaction and so on. The compounds were tested for their vasorelaxant activity in low (30 mmol.L-1) and high (80 mmol.L-1) KCl-induced contraction of rat aorta to identify potential potassium channel openers in vitro. RESULTS: Three series of twenty benzopyran-4-one hydrazone derivatives, nominated N-aminoacetyl-(6-cyano-3,4-dihydrospiro [2H-1-benzopyran-2,1' cyclohexane]-4)-one hydrazone (I), 2-(6-cyano-3, 4-dihydro-2H-1-benzopyran-4 ylene) hydrazinethiocarboxamide derivatives (II) and N-(2-arylethyl) aminoacetyl (6-cyano-3,4-dihydro-2H-1-benzopyran)-4-one hydrazone (III), have been synthesized. They (I1-9, II1-4 and III1-7) are new compounds. Their chemical structures were determined by IR, 1HNMR, MS and elemental analysis. The vasorelaxant effects of those novel compounds indicated that some of the compounds have vasorelaxant activities at 1 x 10(-6) mol.L-1. CONCLUSION: The vasorelaxant activities of compounds I9, III2 and III5 in inhibiting low KCl induced vasocontraction at 1 x 10(-6) mol.L-1 are less potent than the reference compound emakalim. However they are more potent than emakalim to inhibition high concentration KCl-induced vasocontraction at 1 x 10(-5) mol.L-1. It is worthy of further study. PMID- 12567777 TI - [Isolation and structure identification of ligan glycosides from pine needles of Pinus massoniana lamb]. AB - AIM: To study the chemical constituents of the pine needles of Pinus massoniana lamb.. METHODS: Various chromatographic techniques were used to separate and purify. Their physico-chemical properties and spectral data (UV, IR, MS, 1H-1 H COSY, HMQC, DEPT, HMBC and ORD ect.) were measured for structure elucication. RESULTS: Three compounds were isolated from the n-BuOH fraction of water extracts. Their structures were identified as massonianoside A (4), massonianoside A: (7S, 8R)-3, 4, 9'-trihydroxyl-3-methyoxyl-7, 8 dihydrobenzofunan-1'-propanolneoligan-9-O-alpha-L-rhamnopyranoside, massonianoside C (5), (7S, 8R)-9,9'-dihydroxyl-3,3'-dimethyoxyl-7,8 dihydrobenzofunan-1'- propanolneoligan-4-O-alpha-L-rhamnopyranoside and cedrusin 4-O-beta-glucoside (6), (7S, 8R)-3',9,9'-trihydroxyl-3-methoxyl-7,8 dihydrobenzofunan-1'- propanolneoligan-4-O-beta-D-glucopyranoside. CONCLUSION: Compound 4 and 5 are new compounds. PMID- 12567778 TI - [Studies of the chemical constituents of Swertia davida Franch]. AB - AIM: To study the active constituents of Swertia davidi Franch.. METHODS: Chromatography was used to isolate and purify the chemical components, their structures were identified by spectral analysis. RESULTS: Three compounds were identified as 1,7-dihydroxy-3,8-dimethoxyxanthone (gentiacaulein) (V), 1,8 dihydroxy-3,7-dimethoxyxanthone (methylswertianin) (VI) and 1,8-dihydroxy-3,4,7 trimethoxyxanthone (VII). CONCLUSION: Compound VII is a novel xanthone, named daviditin A, the others were isolated from Swertia davidi Franch. for the first time. PMID- 12567779 TI - [Chemical components of Boswellia carterii]. AB - AIM: To investigate the chemical components of Boswellia carterii. METHODS: Chromatographic technologies were used for separation and purification, while spectral analysis was used for structure elucidation. RESULTS: Six compounds were isolated and their structures were identified as acetyl-alpha-boswellic acid (1), acetyl-beta-boswellic acid (2), lup-20(29)-ene-3 alpha-acetoxy-24-oic acid (3), alpha-boswellic acid (4), beta-boswellic acid (5) and acetyl-11-keto-beta boswellic acid (6). CONCLUSION: Compound 3 is a new constituent. PMID- 12567780 TI - [Discussion of the application of three-point interaction principle to the phenylglycine racemic compound in IR spectrometry and its enantiomers separation in HPLC]. AB - AIM: To discuss the relationship between the three-point interaction principle in the stereoselective separation of chromatography and applying this principle to survey the infrared spectrometry of racemate. METHODS: In proving the applicability of the three-point interaction principal in IR spectrometry, a special case was found that phenylglycine did not obtain enantioselective separation on the chiral column but its IR spectrometry still obey this principle and explained such special case by experiment. RESULTS: After an equal quantity of solid crystal of d-phenylglycine and l-phenylglycine were mixed and ground for several minutes, they transformed to racemic compound. X-powder diffraction also confirmed this fact. CONCLUSION: The three-point principle was relatively reliable when it was used in the enantioselective chromatography separation and the IR spectrometric analysis. The reason of the fact that phenylglycine was not separated by chiral column can be explained by the fact that the acting force between the three polarity groups in the enantiomers is so strong that they can not form the instantaneous diastereoisomer with the chiral column, it was agreeable with the phenomenon that racemic mixture easily became racemic compound only by simply grinding the mixture in IR spectrometric experiment. PMID- 12567781 TI - [Quality control for recombinant adenovirus-IL2]. AB - AIM: To investigate the quality and optimized test methods and establish the quality specification of recombinant adenovirus-IL2. METHODS: The titer of Adv IL2 was measured by cytopathic effect (CPE). Hela cells were infected with Adv IL2 in vitro, the expressed IL-2 and its bioactivity in Hela cell were determined by enzyme-linked immunosorbent assay (ELISA) and 3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyltetrazolium bromide (MTT) respectively. The purity of Adv-IL2 was analysed by UV and IE-HPLC method. The molecular weight and enzyme digestive map of Adv-IL2 genome were analysed by electrophoresis. The characteristic gene E2B, IL-2 expression casseter and foreign factors (RCV, HIV, HBV, HCV) were detected with polymerase chain reaction (PCR). Other tests were carried out according to the Chinese Requirements for Biological Products. RESULTS: Adv-IL2 was generated efficiently with a titer of 3 x 10(10) pfu.mL-1. The expressed IL-2 and its bioactivity were 25 ng.mL-1 and 700 u.mL-1 respectively. A260 nm/A280 nm was 1.23. The purity determined by IE-HPLC was higher than 98%. The molecular weight, enzyme digestive map of Adv-IL2 genome, the characteristic gene E2B and IL-2 expression casseter conformed to expected values. CONCLUSION: The specification for Adv-IL2 is established and can be used for the quality control of the product. PMID- 12567782 TI - Studies on the thermal decomposition process and kinetics of purine drugs. AB - AIM: To study the thermal stability, decomposition process and kinetics of such purine pharmaceuticals as aciclovir (Acv), penciclovir (Pcv), and their parent substance, guanine. METHODS: Using infrared technique, accelerating test method and thermogravimetry to investigate the thermal decomposition processes and using Coast-Redfern method, MKN method and Ozawa method to deal with the data to get kinetic functions. RESULTS: The decomposition process and the formed products were derived, the kinetic model function was suggested by comparison of the kinetic parameters. CONCLUSION: Pcv and Acv's degrading product for the first step is guanine. The sequences of their thermal stabilities is: Pcv > Acv. The two drugs' kinetic equation of thermal decomposition is expressed as: da/dt = Ae Ea/RT2(1-alpha)3/2. PMID- 12567783 TI - [The enhancing effect of electroporation and iontophoresis on the permeation of insulin through human skin]. AB - AIM: To study the enhancing effect of electroporation and iontophoresis on the permeation of insulin through human cadaver skin in vitro. METHODS: Using side by side two-chamber diffusion cells, the flux of insulin achieved with iontophoresis and electrophoration were compared. RESULTS: The application of high-voltage pulse combined with iontophoresis resulted in higher flux transdermal permeation of insulin than either one technique alone (P < 0.05). Pulsing at a higher voltage increased the flux of insulin more dramatically than pulsing at a lower voltage (P < 0.01). The transdermal transport of insulin by 90 pulse of 500 V (exponential pulse generater, pulse time: 20-24 ms, pulse frequency: 3 pulse.min 1) followed by iontophoresis led to a quick input and a high steady flux. CONCLUSION: Electroporation combined with iontophoresis can enhance the permeation of insulin significantly. PMID- 12567784 TI - [Preparation of buspirone hydrochloride buccal adhesive tablet and study on its drug release mechanism]. AB - AIM: To prepare buspirone hydrochloride buccal adhesive tablet and investigate factors that influence drug release behavior and the drug release mechanism. METHODS: Buspirone hydrochloride buccal adhesive tablet was prepared with double layers structure composed of drug core and adhesive layer. The materials of the drug core were carbopol 974 and lactose, the adhesive layers were carbopol 974 and HPMC K4M. The influence of drug release factors such as adhesive layer component, adhesive layer weight and adhesive tablet hardness was investigated. The relationship between adhesive layer weight and drug release mechanism in vitro was studied. RESULTS: The results showed that the weight of the adhesive layer and the hardness of adhesive tablet showed significant effects on drug release, but the adhesive layer component showed no significant effect. The optimum prescription of buspirone hydrochloride buccal adhesive tablet was carbopol: HPMC = 1:1, adhesive layer weight = 50%, and adhesive tablet hardness = 4 kg. The study of drug release mechanism from adhesive tablet showed that it was double directions when adhesive layer weight was 20%, and single direction first then double directions when 33.33%, and single direction all along when 50%. CONCLUSION: The speed and direction of drug release from adhesive tablet can be controlled by regulating adhesive layer weight. PMID- 12567785 TI - [Progress in studies on neuroprotective agents]. PMID- 12567786 TI - [Application of membrane-active peptides in the intracellular targeting drug delivery system]. PMID- 12567787 TI - [Advances in research on pharmacogenomics]. PMID- 12567788 TI - Taking a new path. PMID- 12567790 TI - From mini skirts to specialist roles. Interview by Chris Bowles. PMID- 12567791 TI - The price of war. PMID- 12567792 TI - Just rewards. PMID- 12567795 TI - Beating stigma. PMID- 12567796 TI - The greatest gift. PMID- 12567797 TI - The effectiveness of cognitive behavioural therapy in depression. AB - AIM: To examine the process and outcome of the use of cognitive behavioural therapy (CBT) to treat depression. METHOD: The study was undertaken at an acute psychiatric assessment and treatment centre based in the community. The centre operates an open referral system. The study sample was taken from clients attending the centre for mental health assessment who were subsequently diagnosed with unipolar depression. The total sample of 38 clients were allocated to either a treatment group (n = 19) or waiting list control group (n = 19) matching for age, gender and social support. The treatment group was divided into four groups. Each of these groups received 12 sessions of CBT, derived from the procedures described by Beck et al (1979). Data from dysfunctional thought diaries and Beck depression inventory scores were analysed. RESULTS: The results confirm those of previous outcome studies, that CBT is an effective treatment for depression. The results also indicate some interesting points relating to the expression of negative emotions as depression scores improve. It appeared that the expression of sadness preceded the expression of anger. Expression of sadness may be a precursor to expression of anger and necessary for recovery from depression. CONCLUSION: The findings of this study indicate that dimensions of an individual's personality, such as perfectionism and sociotrophy, may be influential in determining the outcomes of CBT. Further research in this area is indicated. PMID- 12567798 TI - Swallowing difficulties protocol: medication administration. AB - Following his research (Wright 2002), David Wright developed this protocol for nurses to use when administering medication to patients who have swallowing difficulties. PMID- 12567799 TI - Evidence-based practice. AB - This article summarises how evidence-based practice is defined, and what might constitute evidence. It describes the steps that should be taken in a systematic review of evidence, and some of the issues involved in implementing the findings of such reviews in practice. PMID- 12567800 TI - Cash backing. PMID- 12567801 TI - [Health and mental health policies in Brazil: social exclusion/inclusion as intention and action]. AB - This study emphasizes the specific characteristics of the mental health as a health policy in the social political field, in Brazil. The objective of this study is to analyze on the Society/State/Health articulations between the structural policies and the specific politic program by means of the historical rescue of these policies, identifying the characteristics and problems at each moment. Presently, an impact between the two strategies of the psychiatric assistance is observed: the hegemonic, hospital centered model which abducts lives, mutilate bodies and minds and trade the Health and, that one, against to the predominant model, searching the rupture by criticizing the Brazilian society. PMID- 12567802 TI - [Infant mortality in a municipality of the interior of the State of Sao Paulo]. AB - Considering that infant mortality indicates the levels of health in the population, we have accomplished the foolwing work, which goals were to identify the causes of infancy mortality during the year of 1998 in Botucatu. The rate of infanty mortality obtained was as much as 12/1000 born alive with greater participation of the neonatal deaths-8,3/1000 born alive. Though most of deaths can be classified as reducible or partially reducible, but the necessary attention to make such reduction possible has varied. All the deaths that have occurred, 21.7% were classified as unavoidable, emphasizing that to reduce the rates of infant mortality, we must continue investing on the quality of health assistance as well as on the improvement of life conditions of the population. PMID- 12567803 TI - [Comparative study on the identification of occupational risks for nursing workers at 2 basic health units of the municipality of Sao Paulo]. AB - Occupational risks of nursing work in Basic Health Units have not been study frequently, so it stimulate us to develop this paper, whose objective are: to compare the perception of nursing workers of two Basic Health Units about these risks and to establish a parallel between the health problems with their exposition to occupational risks. Data were collected by an interview and were analyzed according to their frequencies. Biological risks were observed more frequently. The conclusions show that nursing workers have difficult to understand the genesis of occupational risks. PMID- 12567804 TI - [The informatics process in health: subjects discussed in articles published from 1978 to 1998]. AB - The principal themes are described approached in goods of indexed newspapers, in two bases of available data in Internet, in the period from 1978 to 1998. Consultations were accomplished BIREME, through Internet, being used the descriptors in several orders. A total of 54 goods was obtained that were codified and tabulated. It was ended that, in the studied sample, the thematic of the researches developed on computer science in health had a profile change that passed of theoretical studies for applications in the work atmosphere. PMID- 12567805 TI - [Characterization of a group of hospitalized elderly women and their caretakers keeping in mind the care after hospital discharge]. AB - In the present study 50 old women interned in a medical treatment unity and their respective caregivers were studied. It was verified that most of the women preseted various dependencies and, thErefore, they needed the presence of a caregiver for their survival. The caregivers, most of them female, belonged to the old women's family, had some scholarship degree and pointed out several difficulties en caring for the women. It is verified, consequently, that during hospitalization there is the need to take measures aiming at preparing the caregiver to take on the complex aid required by the old person, especially after hospital discharge. PMID- 12567806 TI - [System of patient classification: application of a validated tool]. AB - The authors applied an instrument of patient classification to the patients of Hall B2 of the Hospital das Clinicas de Marilia, in two distinct periods of 1999, seeking to discover the degree of dependency of the patients in relation to nursing staff. The sample consisted of 31 patients in the first period and 35 in the second. The results showed that most of the patients had been classified as needing minimum or intermediary care in their units of admission. Some modifications of the procedures were suggested. PMID- 12567807 TI - [Iatrogenic events with medications at the intensive care unit: procedures and feelings expressed by nurses]. AB - Iatrogenic occurrences with medication in ICU are adverse events that demand nurse's interventions. The aims of this study were:- to identify nurse's procedures and feelings in this situation;- to characterize factors related to this kind of event in ICU settings. A questionnaire was answered by 148 ICU nurses (76.7%) from 7 hospitals in Sao Paulo city, in 1997 and 1998. The results showed that of 407 mentioned procedures, 31.7% included to communicate the event to the doctor, followed by to intensify the patient control (26.5%). Regarding the 260 feelings informed by the nurses, anxiety and impotence were the most frequent, respectively, 38.1% and 14.6%. Of 303 factors related to the occurrences nurses pointed out neglect (24.6%) and professional inexperience (15.1%) as the most important. Forty three point two per cent (43.2%) of the nurses hardly experimented this situation in their professional life in ICU. PMID- 12567808 TI - [Therapeutic relation with children in the preoperative period: use of play and dramatization]. AB - The purpose of this study was to analyse the therapeutic relationship between a nursing student and a 3 years old child during perioperative period. Through the use of careful development assessments, preoperative tours, and therapeutic play techniques her fears and anxiety because of hospitalization and surgery were reduced and the surgical experience was lived by the child in a constructive manner. Parents and peri-operative team have evaluated positively the preparation and assessment of the child for surgery. PMID- 12567809 TI - [Operative group as a learning strategy in a nursing graduation course: a continent for the fourth year students' experiences]. AB - The objective of this study is to offer a space for the fourth-year students of the nursing school to express their experiences. A space was put in operation by the use of an operative group technique, and the subjects of the study, centered in the task, exchange experiences and evaluate their participation in the groups. To understand the students' experience, a qualitative study with the research action method was produced. The problematic of the study involved the concept that learning is a socialization process which demands changes and mobilize the novices' feelings. The operative group worked as a continent for the students' experiences in which they could explain their feelings. Such feelings, mobilized on their graduation process, were impossible to be ignored the nursing teacher. PMID- 12567810 TI - [Invisible or hidden sexuality in nursing?]. AB - This article is a reflection which pervades the theme about invisibility or hiding of sexuality in the nursing care practice, or besides, of the nurses themselves, leading to questions which hold this bearing, how it was created and what maintains it. The approach presented here permits to assure that this theme goes beyond technical informative itself and, at the same time, is revealed in the nurses's cultural diverseness, giving the opportunity to instrument people for the being and the doing in nurging which provides an experience of life much more qualitatively meaningful. PMID- 12567811 TI - [Alternative/complementary treatments -- the view of nursing students]. AB - The objective of this research was verify if Nursing students know Alternative Practices of Health (A.P.H.), if this knowledge is acquired during the graduation and if they use themselves or recommends this use. The bases were collected near to 96 students of the 4 years of graduation, that were shared out so that is the same as 30% of the students. We verified that 93.73% of the students know A.P.H., that this knowledge is not acquired during the graduation, but principally through the popular sense. Although the most of students (86.46%) recommend the use of A.P.H., principally because they believe in its effectiveness, only about the third part makes use of them. PMID- 12567812 TI - [Awareness education of the diabetic elderly: a nurse's intervention proposal]. AB - The purpose of this study was to identify educative needs of elderly people according to their perceptions elaborating an educative proposal. Data were collected from September 1996 to March 1997 and this sample was formed by 26 elderly patients with diabetes, which were members of the Association of Patients with Diabetes of Uberaba--M.G., Authors utilized the participant observation and semi-structured interview. Data were analysed according to the presupposition of the generator themes proposed by Paulo Freire. The results showed that the Association of Patients with Diabetes aims at their health education. Four generator themes emerged: Diabetes Mellitus type 2, Attention in Health Services, Associated Diseases and the Elderly Patient with Diabetes. PMID- 12567813 TI - [Knowledge of the diabetic patient on foot care]. AB - The objective of this research is to evaluate how much the clients that frequently went to the Diabetes Ambulatory, knew about their own ill, and the feet care. In the sample--diabetes type 2--the majority were people with more than fifty years old and 71.8% were diabetics for more than ten years. So the conclusion that could be taken is that the clients knew about the care that they must have with their feet to don't have complications, but the "own care" is not well done. To insert it in the rightly on their day-by-day they must have access to the information about diabetes, participation of the family, motivation and their own ability. PMID- 12567814 TI - More nursing graduates: an opportunity for rehab? PMID- 12567816 TI - Assistive technology: providing independence for individuals with disabilities. AB - Despite improvements in technology and health care, the number of people with disabilities, and the complexity of needs that they and their families experience, continue to increase. In response to these needs, specialized technology has been developed that helps people with disabilities to become more independent and more involved in the activities in their homes, schools, and communities. However, many individuals with disabilities, their family members, and many rehabilitation service providers are not aware of the availability, use of, and benefits afforded by assistive technology devices. Further, many providers have not received the pre-service academic preparation required to provide services and support to their clients. Preparation includes acquiring the knowledge and skills needed for evaluation of client needs, making appropriate assistive technology recommendations, and developing advocacy skills such as writing letters of necessity that justify funding for assistive technology devices. This article provides information about these issues and suggests that further information is available through continuing education courses as well as articles in the reference list and other sources cited in the accompanying tables. PMID- 12567817 TI - Restorative Nursing Bladder Training program: recommending a strategy. AB - This article describes a Restorative Nursing Bladder Training program, a performance improvement initiative aimed at helping patients regain bladder control by postponing voiding, and then urinating according to a specific, individualized timetable. Data were collected on 14 patients who participated in the program from April 1999 through January 2001. Eight patients achieved a 100% reduction in incontinent episodes; 1 patient documented a 90% reduction; another patient attained an 85% reduction; 3 patients showed a 50% reduction; and 1 patient showed a 30% reduction. PMID- 12567818 TI - Spinal cord injured women's views of sexuality: a Norwegian survey. AB - The purpose of this study was to learn the views of women with spinal cord injuries (SCI) about their sexual life 1 or more years after the trauma. Based on a descriptive survey design, data were collected through a self-administered questionnaire completed by 48 women. The data were analyzed with descriptive statistics. The results show that most of the respondents had been injured 10 or more years. More than 50% were sexually active before the trauma. Fifty percent of the respondents reported that their views on sexuality had not changed as a result of the trauma. Twenty-one percent reported that sexuality was less important to them after the trauma. There is a need for studies of possible nursing interventions that will influence SCI women to recognize both their need for, and their right to, a satisfactory sexual life. PMID- 12567820 TI - Won't you be my neighbor? Mr. Rogers doesn't live here--home visits in New York City. PMID- 12567819 TI - The effect of curriculum on the attitudes of nursing students toward disability. AB - Societal attitudes toward people with differing abilities are often based on a lack of understanding, fear of the unkNown, and stereotypes learned from others. Nursing students enter their educational programs with similar attitudes and experiences. Attitudes of nurses are key to how they respond toward individuals with disabilities and chronic illnesses. The nurses' attitudes affect the individual client's understanding of the disability and his or her self-concept as he or she adjusts to major life changes. The purpose of this descriptive study was to identify whether a change in curriculum and experience had an effect on nursing student attitudes. Using the Attitudes Toward Disabled Persons (ATDP) Scale, a pre-post study of 42 nursing students' attitudes toward individuals with disabilities was conducted. Tests were administered as the students entered their senior year and took the chronic illness course, and again at the completion of the senior year. The students' attitudes were significantly more positive at the completion of their senior year. The findings suggest that education about, and experience with, individuals with disabilities positively affect the attitudes of nursing students toward individuals with disabilities. PMID- 12567821 TI - Assessment scale for caregiver experience with dementia. Development of an instrument in Japan. AB - A great deal of research has been conducted concerning the burden on care-givers of elderly patients with dementia, but only a small number of qualitative studies discuss actual ways individuals with dementia are being cared for and describe the caregiving experience. Scales for assessing caregiver experience have not yet been developed. The aim of this study was to develop an instrument on the basis of the caregiver experience model developed by Kobayashi [Suwa], Masaki, and Noguchi (1993) in Japan. Psychometric evaluations of the scale were conducted. A sample of 116 caregivers were asked to fill out a questionnaire on a self-rating 5-point scale measuring their sense of caregiving burden, and 90 responses were obtained. The data measured with this original scale was subjected to factor analysis and an assessment scale was developed. As a result, the Assessment Scale for Caregiver Experience with Dementia (ASCED) was created. It consists of 24 items, divided into three subscales--the empathetic caregiving experience, the disciplinary caregiving experience, and the resigned caregiving experience. It also divides caregivers into six types. Work needs to be performed not only to further evaluate changes in the caregiving experience through refinement of the ASCED, but also to qualitatively and quantitatively investigate the nature of the care given to elderly individuals with dementia. PMID- 12567822 TI - Enhancing relationships between care providers and residents in long-term care. Designing a model of care. AB - Research in three different areas was examined and several conclusions can be drawn. Continuity of care provider is critical to understanding the resident and to developing relationships with the resident over time (Patchner, 1987; Teresi et al., 1993a). The teaching of interactional skills is not enough; the care provider must be engaged in some way, such as learning about the person through life stories (Best, 1998; Caris-Verhallen et al., 2000; Coker et al., 1998; Heliker, 1999; McCallion et al., 1999; Pietrukowicz & Johnson, 1991; Williams & Tappen, 1999). If care providers are called to enhance relationships with residents, they too must be supported in their work environments (Hallberg & Norberg, 1993; Montegar et al., 1977; Kovach & Krejci, 1998). Finally, research confirmed positive secondary resident and care provider outcomes can be achieved following the development of holding relationships. Overall, preliminary empirical support for the capacities of the care provider--reliability, empathy, continuity--and for the requirement for support were established from a review of the literature. However, no intervention studies were found that incorporated the complete set of theoretical variables. Testing the combined influence of these variables, as conceptualized by Winnicott's (1970) theory of relationships, and the manner in which they affect the holding relationship for residents, and subsequently secondary care provider and resident outcomes, is essential to assess the usefulness of this theory to relationship building in LTC. Caregiving relationships involve all kinds of social interaction during the course of which the patient's sense of self-worth can either be enhanced or thwarted (Agich, 1990). Therefore, a milieu should be developed to accentuate care provider resident relationships and lead to a systematic and encompassing framework of positive expectations on the part of all nursing personnel involved. A model of care focusing on relationships may be one means to this end. PMID- 12567823 TI - Self-transcendent views and behaviors. Exploring growth in caregivers of adults with dementia. AB - Literature suggests difficult life experiences such as fear, loss, and grief can provide an impetus for moving from self-centeredness toward a broader personal identity and self-transcendence. Caring for a family member with dementia may provide the context for such a movement; thus, the purpose of this study was to investigate self-transcendent views and behaviors in family caregivers of adults with dementia. A naturalistic field study design was used to collect data from individuals caring for family members with dementia. Findings showed little evidence of self-transcendent views and behaviors and little opportunity for caregivers to engage in activities that might facilitate self-transcendence. Conclusions of the study reveal social isolation, ambivalence, burden of care, and emotional fragility may inhibit the development of self-transcendence and interfere with healthy growth and development in family caregivers. PMID- 12567824 TI - Dehydration in terminally ill patients. Perceptions of long-term care nurses. AB - Dehydration in terminally ill patients has been found to be beneficial and to improve the quality of an individual's last few days of life. As the population continues to age, more individuals are cared for in long-term care (LTC) facilities, where they tend to spend their final days. Previous studies have examined the perceptions and attitudes of hospice nurses, acute care nurses, physicians, and caregivers; however, no such studies have evaluated LTC nurses. It is necessary to know LTC nurses' perceptions and attitudes so they can be offered the education needed to provide the best quality care for terminally ill patients. The purpose of this study was to describe how nurses working with elderly individuals in LTC perceived terminal dehydration (TD). Long-term care nurses (N = 64) were surveyed using a modified version of an established 10-item instrument. Significant findings included a positive correlation between age and positive perception of TD--as nurse age increaSed, a more positive view of TD was expressed. Also, the number of deaths witnessed was positively associated with the belief that TD was beneficial. In general, responses to the individual survey items were quite varied, representing inconsistencies in attitudes and care of dying LTC patients. The results of this descriptive study indicate the debate concerning the benefits of TD continues and remains an important topic for the LTC nurse. PMID- 12567825 TI - Coping with anxiety in later life. AB - The goal of this study was to determine how older adults cope with three forms of anxiety, and potential avenues for applied interventions. Although the findings shed light on some interesting findings with potential psychosocial applications, several limitations need to be noted. First, this study was based on two assumptions. The assumption, based on earlier work (Carver et al., 1989; Lazarus & Folkman, 1984; Zeidner & Saklofske, 1996), that certain coping strategies are more effective than others, and an assumption of the direction of influence in which anxiety is a precursor of coping strategies. Because this was an exploratory study, the research questions did not directly test these assumptions. Second, this study is correlational in nature. Therefore, conclusions cannot be drawn about the causality of these associations. Third, as with any self-report data and self-selected sample, one needs to interpret the findings with caution. Similarly, for the purposes of the study, a non-clinical sample of older adults was examined using three distinct conceptualizations of anxiety. Suggestions for future research include: Replication of this study using a multidimensional measure of anxiety appropriate for clinical samples. A longitudinal replication of this study identifying patterns of coping that facilitate adjustment over time. Finally, a more general purpose of this study was to focus attention on a neglected issue in gerontology--the experience of anxietY in later life (Frazier & Waid, 1999; Gatz, 1995; Rabins, 1992; Shamoian, 1991; Sheikh, 1992; Smyer, 1995; Stanley & Beck, 1998), and, most importantly, the role of gerontological nurses in early assessment and intervention for successful treatment of anxiety in older adults. PMID- 12567827 TI - When your patient is hearing impaired. PMID- 12567828 TI - Spina bifida. PMID- 12567829 TI - Nurse as patient. A fast-paced life takes its toll. PMID- 12567830 TI - Organ donation: a delicate balance. PMID- 12567831 TI - Triaging lower abdominal pain. PMID- 12567832 TI - Perineal care products. PMID- 12567833 TI - What if you're named in a lawsuit? PMID- 12567836 TI - [Listeriosis and the pregnant women]. PMID- 12567835 TI - Shortages. Reinvesting in nursing: an urgent need. PMID- 12567837 TI - [Home care in the context of ambulatory care reform]. PMID- 12567838 TI - [Living with asthma. (Part 1) Evaluation of the asthmatic crisis]. PMID- 12567839 TI - [For a practice based on convincing results]. PMID- 12567840 TI - [Perinatal interventions. The importance of convincing data]. PMID- 12567841 TI - [Reforms and restructurings in nursing care. The nurse in the heart of the torment and the solution]. PMID- 12567842 TI - [Advanced practice in neonatology]. PMID- 12567844 TI - Volunteer opportunities for registered nurses. PMID- 12567845 TI - DRG Validation Advisory Committee (DVAC). PMID- 12567846 TI - The new nurse and the nursing shortage. PMID- 12567847 TI - Role of APRNs in palliative care. PMID- 12567848 TI - NDNA Nursing Education Council member responds to "advanced practice nurses role" position statement. PMID- 12567849 TI - Patient education on pain management. PMID- 12567852 TI - The golden anniversary of the Oklahoma Baptist University School of Nursing. PMID- 12567854 TI - Clinical evaluation: an update. PMID- 12567855 TI - Effects of dimethyl diphenyl bicarboxylate on the metabolism and hepatotoxicity of aflatoxin B1 in rats. AB - AIM: To study the effect of antihepatitis drug, dimethyl diphenyl bicarboxylate (DDB) on the metabolism and hepatotoxicity of aflatoxin B1(AFB1) in rats. METHODS: Rats were given orally DDB 300 mg.kg-1.d-1 for 3 days and then injected intraperitioneally with AFB1 1.5 mg.kg-1. Serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were examined 16 hours after the injection of AFB1. The in vitro metabolism of AFB1 by DDB-pretreated rat liver microsome was investigated by HPLC assay. RESULTS: DDB (300 mg.kg-1) pretreatment provided significant protection against AFB1 hepatotoxicity as evidenced by the decrease of AFB1-elevated serum marker enzymes in rats. Pretreatment with DDB was shown to slightly increase the level of AFM1, the less toxic metabolite. DDB significantly increased the liver cytochrome P450 content, P450 isozyme 2B1-mediated 7-pentoxyresorufin O-dealkylase (PROD) activity, cytosolic glutahione (GSH) level and GSH S-transferase (GST) activities. In addition, DDB slightly increased P450 isozymes, 3A-mediated erythromycin demethylase and 1A-mediated 7-ethoxyresorufin O-deethylase (EROD) activities. CONCLUSION: The results indicate that DDB protected rats against AFB1 hepatotoxicity by increasing the detoxifying metabolism of AFB1 in the liver. PMID- 12567856 TI - [Recombinant human brain natriuretic peptide on the cardiac hemodynamics and renal function in dogs with heart failure]. AB - AIM: To study the effects of rhBNP on the cardiac hemodynamics and renal function in dogs with heart failure. METHODS: Congestive heart failure in dogs was induced by either rapid ventricular pacing (RVP), 250 beats.min-1 for 7-14 days or by thoracic inferior vena cava constriction (TIVCC) to 1/2 its original diameter. When remarkable hemodynamic changes appeared rhBNP was infused intravenously at the dosage of 10, 30 and 100 ng.kg-1.min-1, each dose lasting 30 min. RESULTS: In dogs (n = 7) with RVP heart failure, intravenous infusion of rhBNP at 10-100 ng.kg-1.min-1, caused decreases in mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), LVdP/dtmax, pulmonary arterial pressure (PAP), left ventricular end diastolic pressure (LVEDP), total peripheral vascular resistance (TPR) and renal vascular resistance (RVR) dose-dependently, without significant changes in cardiac output (CO), LVdp/dt/P, left ventricular work (LVW), renal blood flow (RBF) and heart rate (HR). This suggested that rhBNP reduced the pre load and after-load of the dogs with congestive heart failure but showed no distinct effect on the contractility of the heart. In dogs (n = 7) with TIVCC heart failure, there were remarkable decreases in MAP and LVEDP following the rhBNP infusion, without further reduction of CO, but no marked change in HR, LVSP, LVdP/dtmax, RAP and TPR. In both animal models of heart failure, there were significant increases in urine volume and sodium excretion which were more significant in TIVCC dogs than in RVP dogs. CONCLUSION: rhBNP reduced the pre load and after-load in dogs with heart failure and showed remarkable diuretic effect, but did not affect the contractility of the heart. PMID- 12567857 TI - [Studies on antitumor activity of rhEndostatin]. AB - AIM: To investigate the inhibitory effect of recombinant human endostatin (rhEndostatin) on endothelia cell proliferation and tumor growth. METHODS: MTT assay was applied to examine the anti-proliferation of rhEndostatin on human embryo umbilical cord vascular endothelial cell ECV304 and human cancer cell HCT 8, BGC803 and EJ. Xenotrasplanted nude mice models with human cancer and experimental implanted tumor mice model were used to evaluate rhEndostatin's antitumor activity. RESULTS: rhEndostatin was shown to inhibit the proliferation of ECV304 cells and the IC50 is about 7 x 10(-6) g.L-1. No inhibition was observed in HCT-8, BGC803 and EJ cells at 1 x 10(-4) g.L-1 rhEndostatin. rhEndostatin was shown to inhibit human xenograft in nude mice with human gastric cancer BGC803 and breast cancer B37 when administered subcutaneously at 5, 10, 20 mg.kg-1.d-1 for 24 days in a dose-dependent manner. Mouse hepatoma H22 was also suppressed when given rhEndostatin subcutaneously 20 mg.kg-1.d-1 for 9 days, but it showed no inhibitory effect on Lewis lung carcinoma and B16 melanoma. CONCLUSION: These results indicate that rhEndostatin can inhibit the growth of xenotransplanted human tumors in nude mice and certain murine tumor. The action mechanisms may be that it can inhibit endothelial cell proliferation, thereby inhibiting the formation of new blood vessel in tumor, leading the tumor to stop grow. PMID- 12567858 TI - [Effects of leflunomide and its active metabolite on the production and mRNA expression of TNF-alpha in peritoneal macrophages and synovial cells with adjuvant arthritis in rats]. AB - AIM: To observe the effects of leflunomide (LEF), an isoxazole immunomodulatory agent and its active metabolite, A771726, on the production and mRNA expression of TNF-alpha in peritoneal macrophages and synovial cells with adjuvant arthritis rats and to further investigate the immunosuppression effects of leflunomide and its mechanisms. METHODS: ELISA methods were used for assaying the levels of TNF alpha. The RT-PCR methods were used for measuring the expression of TNF-alpha. RESULTS: The production of TNF-alpha was increased in the supernatant of PM psi in adjuvant arthritis (AA) model rat. LEF (5, 10, 25 mg.kg-1, ig) was shown to inhibit the release of TNF-alpha from peritoneal macrophages induced by LPS and the inhibitory effects were in a dose-effect relevance manner. A parallel investigation of cytokine mRNA expression was undertaken using semi-quantitative reverse transcribed polymerase chain reaction (RT-PCR) to follow the kinetics of cytokine appearance in PM psi and synovial membrane tissue cells obtained from AA/normal rats treated with A771726. The results of RT-PCR from macrophages and synovial membrane tissue cells of AA rats at the peak of inflammatory phase showed that TNF-alpha mRNA expression levels were higher than those of normal rats, while the expression of TNF-alpha mRNA was reduced by treating with A771726 in vitro. On the other hand, the TNF-alpha mRNA expression showed kinetics very similar to those obtained by ELISA technique which measured protein expression. CONCLUSION: Leflunomide and its active metabolite, A771726, was found to inhibit the production and mRNA expression of TNF-alpha in peritoneal macrophages and synovial cells with adjuvant arthritis rats model. It might be involved in the mechanisms of its anti-inflammation and immunosupression. PMID- 12567859 TI - C-mycprotein expression upregulated by 2-(3-estrone-N-ethyl piperazine-methyl) tetracycline in bone. AB - AIM: To study the effect of XW630 on expression of pro-oncogene c-myc in the long bones of fetal mice in vitro for postulating the mechanism by which XW630 exerts its effect on bone. METHODS: The fetuses of pregnant mice were removed on day 16 of gestation, the long bones of the forelimbs of female fetal mice were freed of muscle and soft tissue and cultured in a specific device for 48 h in BGJb medium treated with 1 x 10(-7), 1 x 10(-8) and 1 x 10(-9) mol.L-1 XW630 in the final medium. After cultured for 48 h, the long bones were harvested and immunohistochemical analysis was performed for determination of c-Myc protein expression in epiphyseal plates. The areas of positive cells in the resting zone, proliferative zone and hypertrophic zone in epiphyseal plate were determined under image analytic system. RESULTS: When the concentration of XW630 in the medium was 1 x 10(-9) mol.L-1, the area of c-Myc positive cells increased in the proliferative zone compared with 1 x 10(-9) mol.L-1 in the estrone group, significant increase was also observed in the resting zone compared with the control group. When the concentration of XW630 in medium was 1 x 10(-8) or 1 x 10(-7) mol.L-1, stronger expression than that in the control group and the estrone group at the same concentration was observed in each of the three zones. CONCLUSION: The estrogenic effect of XW630 on bone was stronger than that of estrone. XW630 may promote proliferation and differentiation of chondroncytes by promoting c-Myc protein expression in chondroncytes. Thus, endochondral bone formation was enhanced. PMID- 12567860 TI - [Syntheses and antitumor activities of the derivatives of P-methyl goniotriol in vitro]. AB - AIM: To find derivatives of p-methyl-goniotriol with more potent antitumor activities and lacking undesirable effects. METHODS: Eighty derivatives of p methyl-goniotriol have been synthesized in nine steps from alpha-D-glucoheptonic delta-lactone (2). Compound 2 reacted with acetone in the catalyzer, H2SO4 and anhydrous MgSO4, and then reacted with 60% aqua HOAc, finally was oxidized by NaIO4 at room temperature into aldehyde 3 in a yield of 71.3%. The aldehyde 3 reacted immediately with P-CH3-PhMgBr giving mixture 4. The mixture 4 was oxidized by NaIO4, reacted with Ph3P = CHCO2Et and then induced by catalyzer. 1,8 Diazabicylclo[5, 4, 0]undec-7-ene in THF providing the compounds 6 and 7. The esterfication of 6 with cinnamyl chloride et al in 4-dimethylaminopyridine, Et3N gave the esters 8a-h. Acid hydrolysis of the acetone protecting group of 8a-h in 75% aqua HOAc gave compounds 9a-h. Their chemical structures were confirmed by IR, 1HNMR, MS and element analysis. The antitumor activities of the compounds were screened by MTT methods. RESULTS: Fifteen derivatives of p-methyl-goniotriol (7, 8b-h, 9b-h) are new compounds. CONCLUSION: Pharmacological tests indicate that these compounds showed antitumor activities toward tumor cells (A2780, HCT 8, Bel742, KB) in vitro. The antitumor activities of 8b, 8d, 8g and 9h were more potent than howiinol A. PMID- 12567861 TI - [Study on metabolites on aconitine in rabbit urine]. AB - AIM: To identify the main metabolites of aconitine in the urine of rabbits. METHODS: After oral administration of aconitine (5 mg.kg-1), the urine of male rabbits was collected and extracted by solid phase extraction and analyzed by liquid chromatography-ion trap mass spectrometry. RESULTS: Aconitine and 4 metabolites were found in the rabbit urine. Their protonated molecular ions at m/z 632, m/z 604, m/z 590, m/z 500 and multistage fragment ions with neutral loss of 60 u, 32 u, 28 u and 18 u were monitored. Their relative concentration were M1 > Aconitine > M4 > M2 > M3. CONCLUSION: The metabolites M1-M4 were deduced as 16 O-demethylaconitine, benzoylaconine, 16-O-demethylbenzoylaconine and aconine, respectively. PMID- 12567862 TI - [Studies on the chemical constituents of Spatholobus suberectus Dunn]. AB - AIM: To study the chemical constituents of Spatholobus suberectus Dunn. METHODS: The constituents of Spatholobus suberectus were separated with various chromatographic techniques. The structures were elucidated by physico-chemical properties and spectral data. RESULTS: Eight compounds were isolated from Spatholobus suberectus Dunn. They were identified as: suberectin (1), formononetin (2), daidzein (3), calycosin (4), pyromucic acid (5), 1,3,5 benzenetriol (6), succinic acid (7) and beta-sitosterol (8). CONCLUSION: Suberectin is a new compound and characterized as 7,3',4'-trihydroxy-6-methoxy flavanone on the basis of chemical and spectral evidence. Compounds 4, 5, 6 and 7 were obtained from Spatholobus genus for the first time. PMID- 12567863 TI - [Structures and pharmacological activities of beesiosides O and P]. AB - AIM: To investigate the chemical constituents of the rhizomes of Beesia calthaefolia native to China in order to obtain a more comprehensive understanding of its effective components. METHODS: Compounds were isolated by column chromatography with silica gel. Their structures were elucidated by spectral analysis and chemical evidence. Compounds identified were subjected to pharmacological evaluation. RESULTS: Two novel compounds were isolated and identified as (20S, 24S)-15 alpha-acetoxy-16 beta, 24; 20, 24-diepoxy-9, 19 cyclolanostane-3 beta, 25-diol-3-O-beta-D-xylopyranoside (I) and (20S, 24R)-15 alpha-acetoxy-9, 19-cyclolanostane-3 beta, 16 beta, 20, 24, 25-pentaol-3-O-beta-D xylopyranoside (II), named beesioside O and beesioside P. CONCLUSION: Compounds I and II are new compounds. Compounds I exhibited immunosuppressive activity and could inhibit angiogenesis as well as inhibit the proliferation of osteoblast. Compound II displayed remarkable inhibition activity against calcium channel receptor. PMID- 12567864 TI - Chemiluminescence flow-through sensor for the determination of vitamin B2 using controlled-reagent-release technology. AB - AIM: To establish a simple and novel method for the determination of vitamin B2 rapidly in pharmaceutical preparations. METHODS: Vitamin B2 was determined by a chemiluminescence (CL) sensor combined with flow-injection (FI) technology. The analytical reagents involved in the CL reaction, luminol and hexacyanoferrate (III), were both immobilized on an anion-exchange resin column in FI system. The CL signal produced by the reaction between luminol and hexacyanoferrate (III), which were eluted from the column through sodium phosphate injection, decreased in the presence of vitamin B2. RESULTS: The decreased CL intensity was linearly correlated with the vitamin B2 concentration in the range of 0.01-1.0 microgram.mL-1, the detection limit was 4.0 ng.mL-1 vitamin B2 (3 sigma). At a flow rate of 2.0 mL.min-1, the procedure including sampling and washing could be performed in 2 min with a relative standard deviation of less than 3.0%. CONCLUSION: The flow sensor exhibited both good sensitivity and stability. It could be reused more than 450 times and has been applied successfully to the analysis of vitamin B2 in pharmaceutical preparations. PMID- 12567865 TI - [Determination of six effective components in Rheum by cyclodextrin modified micellar electrokinetic chromatography]. AB - AIM: To determine six effective components (aloe-emodin, rhein, emodin, rhaponticin, physcion and chrysophanol) in Rheum. METHODS: Using buffer solution containing 20 mmol.L-1 borax, 20 mmol.L-1 sodium deoxycholate (SDC), 20 mmol.L-1 sodium taurocholate (STC), 15 mmol.L-1 beta-cyclodextrin (beta-CD) and O-phthalic acid as the internal standard, the six components were determined by cyclodextrin modified micellar electrokinetic chromatography. RESULTS: In less than 25 min, aloe-emodin, rhein, emodin, rhaponticin, physion and chrysophanol were separated. The separation conditions were optimized by adjusting buffer pH, concentrations of SDC, STC and beta-CD. The linearity ranges of aloe-emodin, rhein, emodin, rhaponticin, physcion and chrysophanol were 4-34, 5-40, 4-60, 5-80, 6-90 and 5-85 micrograms.mL-1 respectively. Relative standard deviation (RSD) of the method was less than 2.2%. The recoveries of aloe-emodin, rhein, emodin, rhaponticin, physcion and chrysophanol were 100.0%, 98.3%, 100.4%, 94.6%. 95.2% and 93.8% respectively. Raw Rheum, Mongolian Rheum and Rheum tanguticum samples were analyzed. CONCLUSION: This method can be an effective one for identification of Rhubarb. PMID- 12567866 TI - [Identification of the metabolites of penehyclidine hydrochloride raceme in rats by LC-MS/MS and ion cluster]. AB - AIM: To study the metabolites of penehyclidine hydrochloride (PH) raceme, a new anticholinerigic drug invented by the Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences. METHODS: Three healthy rat urine samples were collected within 24 h after a single i.m. dose of PH raceme and PH-d5 [(5 + 5) mg.kg-1] simultaneously. The eight metabolites of PH raceme were identified by the methods of LC-MS/MS, GC-MS, FAB-MS and the stable isotope ion cluster. Mass spectrometry was operated in the positive mode for the method of LC-MS/MS. RESULTS: M1 and M1* were identified as the oxygenated products of PH in the cyclopentyl group; M2 and M2* were as the hydroxylated products of PH in the cyclopentyl group; M3 and M3* were as the oxygented and hydroxylated products of PH at the meta-position of cyclopentyl group; M4 and M4* were identified as the dihydroxylated metabolites of PH, the hydroxylated position were at the cyclopentyl group and quiniuclidinol ring of PH. Among them, M1 and M1*, M2 and M2*, M3 and M3*, M4 and M4* were the isomers of each other. CONCLUSION: These characteristics can be used for future structure elucidation in studies of the metabolites of PH optical isomers. The structure data of PH metabolites provide important information for the clinical use and for developing better anticholinerigic drug. PMID- 12567867 TI - [Quality control methods for recombinant human endostatin]. AB - AIM: To establish the quality control methods for recombinant human endostatin. METHODS: Biological activity was determined by endothelial cell migration assays. Peptide mapping was tested by trypsin digestion and RP-HPLC. Purity was determined by non-reduced SDS-PAGE and RP-HPLC. Other tests including molecular weight, isoelectrical point, etc. were done according to the National Requirements for Biological Products (2000). RESULTS: The method of bioassay was established and used for determining activity of endostatin. Specific activity of the three batchs of drug substance was 1.45 x 10(6), 1.57 x 10(6) and 2.73 x 10(6) u.mg-1 proteins. Peptide mappings of the three batches of drug substance were completely identified. Both purity results of the products tested by SDS PAGE and RP-HPLC were more than 99%. CONCLUSION: The established methods can effectively control the quality of recombinant human endostatin. PMID- 12567869 TI - [Inhibition of oxygen consumption by annonaceous acetogenins in liver cell respiration and their structure-activity relationship]. AB - AIM: To study the inhibition of oxygen consumption by annonaceous acetogenins (ACG) and their structure-activity relationship (SAR). METHODS: The inhibition of oxygen consumption in chicken liver cell respiration by different structural ACG was studied by using oxygen electrode technique. RESULTS: Six ACG showed potent inhibitory effects like rotenone which was a classical inhibitor of mitochondrial complex I, and was more potent than complex IV inhibitor KCN. The IC50 values of six ACG for inhibiting oxygen consumption suggested that bistetrahydrofuran (THF) ACG was 7-11 times more active than non-THF ACG, and A1-type ACG was more potent than A2-type ACG. CONCLUSION: The terminal gamma-lactone was crucial for the inhibition of oxygen consumption. The distance between THF and gamma-lactone, the hydroxyl groups in the alkyl chain, were the important factors of SAR, but the 4 OH group possibly played some negative role in the exhibit of potent activity. PMID- 12567868 TI - [Studies on heart-protecting musk pH-dependent gradient-release pellets]. AB - AIM: To prepare heart-protecting musk pH-dependent gradient-release pellets and investigate the drug release in vitro and in vivo. METHODS: The pH-dependent gradient-release pellet system was prepared by using HPMC, Eudragit L-30D-55 and Eudragit L100-Eudragit S100 (1:5) combinations as coater. The release of borneol and total ginsenoside from pH-dependent gradient-release pellets were determined according to the method of Pharmacopoeia of the People's Republic of China (2000) in the simulated gastrointestinal pH conditions. The gastrointestinal transit and disintegration of pellets was investigated by using gamma-scintigraphic trace in volunteers. The pharmacokinetics of borneol of heart-protecting musk pH-dependent gradient-release pellets was studied in 6 healthy volunteers by GC methods. RESULTS: The f2 value of release data of borneol and total ginsenoside of the heart-protecting musk pH-dependent gradient-release pellets was 79.6 in the simulated gastrointestinal pH conditions. The gamma-scintigraphic trace evaluation demonstrated that the pellets coated with HPMC, Eudragit L-30D-55 or Eudragit L100-Eudragit S100 (1:5) combinations can disintegrate in stomach, duodenum and jejunum or ileum. The gastrointestinal transit time of pellets was about 5 hours in fasted state and about 6 hours in fed state. The concentration time curves of borneol of heart-protecting musk pills fit in two-compartment model. The pharmacokinetics data showed that borneol had a short time of absorption and elimination. The mean residence time (MRT) of borneol of heart protecting musk pills was 2.61 hours. The plasma concentration of borneol of heart-protecting musk sustained-release capsule which consisted of three kinds of pellets coated with HPMC, Eudragit L-30D-55 or Eudragit L100-Eudragit S100 (1:5) combinations was steadier than those of heart-protecting musk pills, its Cmax was lower than and Tmax was near to those of heart-protecting musk pills, its MRT was 4.0 hours, and its relative bioavailability was 96%. CONCLUSION: The lipidsoluble borneol and watersoluble total ginsenoside of heart-protecting musk pH-dependent gradient-release pellets can release simultaneously while sustained-releasing in vitro. The heart-protecting musk pH-dependent gradient-release pellets had the characteristics of pH-dependent gradient-releasing and disintegration while transiting in gastrointestinal tract. A characteristic of gradient sustained release was shown in the concentration-time curves of borneol of heart-protecting musk sustained-release capsule in volunteers. PMID- 12567870 TI - Recent progress in structure-activity relationship studies on the anticancer drug colchicine and its analogues. PMID- 12567871 TI - [Proteomics in post-genome era and its application in pharmacy]. PMID- 12567872 TI - HIV testing for all nurses is unlikely. PMID- 12567873 TI - Patient power. PMID- 12567874 TI - Smallpox: the disease and strategies for its control. AB - Smallpox was once one of the most severe infectious diseases and was endemic throughout the world for most of recorded history. It no longer exists in nature, having been declared extinct in 1980 following a global eradication campaign led by the World Health Organization. However, recent news reports on the vaccination of key health workers in the event of deliberate release of smallpox as part of a biological attack, and the deployment of Regional Smallpox Diagnosis and Response Teams, are stimulating demand for knowledge of the disease. This article provides an overview of the smallpox virus, including signs and symptoms, treatment and transmission, and vaccination and control in the event of deliberate release. PMID- 12567875 TI - Pelvic inflammatory disease. PMID- 12567876 TI - Using a hand-held Doppler. PMID- 12567877 TI - Erectile dysfunction: assessment, treatment and prescribing issues. AB - Because of the sensitive nature of erectile dysfunction, epidemiology may be based on conservative estimates. Today, oral medication is regularly prescribed, often in the primary care setting, although this may be at the expense of accurate patient assessment. Government restrictions on ED treatment are an obstacle for patients seeking help and may result in their financing themselves. PMID- 12567878 TI - Speeding up the diagnosis of oesophago-gastric cancer. AB - National referral guidelines have been developed for patients with suspected oesphago-gastric cancer with the aim of reducing the delay from presentation to referral and, therefore, reducing overall delay in diagnosis. This study evaluates the impact of these guidelines and shows that they have resulted in a significant decrease in the time from referral to endoscopy for patients. Nurses can further contribute to this by increasing patient awareness of symptoms. A course is being developed to provide the necessary knowledge. PMID- 12567879 TI - Relieving the pressure. PMID- 12567880 TI - Children in need. PMID- 12567881 TI - [Mycoplasma synoviae-associated amyloid arthropathy in white leghorns: case report]. AB - Under field circumstances amyloid arthropathy was nerve recorded in white layers, while experimentally their brown counterparts were found to be more susceptible to the induction of amyloid arthropathy, although articular amyloid was found in a few white leghorns. In the present manuscript the first field case of amyloid arthropathy in white layers associated with Mycoplasma synoviae is reported. In the same house where the white birds were housed, brown layers were present. The condition was much more severe in the latter chickens. The different susceptibility between both breeds is discussed in view of previously performed research. PMID- 12567882 TI - [Eight years of amyloid research in chickens: a review]. AB - The articular localisation of amyloid is not uncommon in Galliformes. In commercial brown layer chickens amyloid artropathy was first described in 1994 as a clinical problem with increasing importance associated with growth depression and lameness. Considering the clinical veterinary and the scientific relevance of amyloid artropathy, it was a subject of poultry research during the past eight years. The outcome of the various studies performed is described in the present update. The occurrence, nature, pathogenesis and possible etiological factors of chicken amyloid artropathy, the identification and characterization of the chicken joint amyloid protein and the gene involved are reported. Finally, the results of immunological studies, which provide a plausible explanation for the difference in breed susceptibility, and summarising conclusions are presented. PMID- 12567883 TI - [The next patient is a sick fish, what to do? (part I)]. AB - Many health problems in fish are the result of living in poor environmental conditions. The clinical history is very important, not only of the fish, but also of the water quality. Watertesting is presented and after making the clinical examination, a diagnosis can be made. In the second article therapy and surgical procedures are discussed. PMID- 12567884 TI - [Henk Jan Ormel: "Staying alert so that the veterinary infrastructure does not disappear through budget cuts"]. PMID- 12567885 TI - [Malfunctioning care of a 'pot-bellied' pig]. PMID- 12567886 TI - [Stay informed and don't jeopardize yourself!]. PMID- 12567887 TI - [Veterinary jurisprudence must leave the ivory tower]. PMID- 12567888 TI - [Studies on evodiamine induced HeLa cell apoptosis]. AB - AIM: To study the mechanism of evodiamine-induced growth inhibition of HeLa cells. METHODS: HeLa cells viability and the effect of caspase inhibitors on evodiamine-induced apoptosis were measured by crystal violet assay. Changes in cellular morphology were observed by phase-contrast microscopy. Apoptosis specific nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. RESULTS: Evodiamine was found to inhibit HeLa cell growth in dose- and time-dependent manners. Caspase-3 inhibitor, z-Asp-Glu-Val-Asp-fmk (z DEVD-fmk) was shown to partially inhibit evodiamine-induced apoptosis. However, caspase-1 inhibitor, Ac-Tyr-Val-Ala-Asp-chloromethyl-ketone (Ac-YVAD-cmk), did not antagonize evodiamine induced cell death. CONCLUSION: Evodiamine suppresses the growth of HeLa cells in vitro by apoptosis. Evodiamine-induced apoptosis is partially dependent on caspase-3 pathway in HeLa cells. Other apoptotic pathways might be also related to the induction of apoptosis by evodiamine. PMID- 12567889 TI - [Sodium selenite-induced oxidative stress and apoptosis in human acute promyelocytic leukemia NB4 cells]. AB - AIM: To explore sodium selenite-induced oxidative stress and apoptosis in human promyelocytic leukemia NB4 cells. METHODS: The growth inhibition of NB4 cells was measured by MTT test. Apoptosis was determined morphologically by Giemsa stain and by DNA ladder formation in electrophoresis. Quantitation of apoptosis was determined by percentage of PI stained cells containing subdiploid amount of DNA measured by flow cytometry. Generation of reactive oxidative species (ROS) in NB4 cells was determined by lucigenin dependent chemoluminescent (CL) test. Spectrophotometer was used to measure the level of reduced glutathione, superoxide dismutase (SOD) and glutathione peroxidase in the cell. RESULTS: Sodium selenite was shown to inhibit the growth of NB4 cells. Sodium selenite induced apoptosis with dose and time dependency: the ratio of subdiploid cells in control group was 1.3% +/- 0.7%. The 5 mumol.L-1 group was 10.4% +/- 1.4%, 10 mumol.L-1 group was 16% +/- 1%, and the 20 mumol.L-1 group was 27.3% +/- 0.8%. Sodium selenite (> or = 5 mumol.L-1) enhanced the ROS level markedly in NB4 cells (in 20 mumol.L-1 group ROS level was increased by 17 times, compared with control group), accompanied with decrease of reduced intracellular glutathione. These effects were time and dose dependent. N-acytlcysteine as an antioxidant was found to inhibit sodium selenite-induced oxidative stress and apoptosis in NB4 cells. CONCLUSION: Sodium selenite can induce apoptosis of NB4 cells and would possibly be used as an agent for the treatment of malignancy. The main mechanism of action might be related to oxidative stress induced by sodium selenite, thereby, leading to apoptosis as shown in NB4 cells. PMID- 12567890 TI - [Effect of melatonin on learning and memory impairment induced by aluminum chloride and its mechanism]. AB - AIM: To investigate the effect of melatonin on learning and memory impairment in mice induced by aluminum chloride and its possible mechanism. METHODS: Mice were treated with intracerebroventricular (icv) injection of 2 microL 5% aluminum chloride solution, once a day for 5 d. At the same time, the mice were given intraperitoneally melatonin 0.6, 3 and 15 mg.kg-1, once a day for 14 d. The passive avoidance of the mice was assessed by step-through test on day 15 after the last icv injection, and then the place navigation and spatial probe ability by Morris water maze were tested. After the spatial probe test, the activities of total superoxide dismutase (T-SOD), CuZn superoxide dismutase (CuZn-SOD), glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA) in the cerebral cortex and hippocampus of mice brain were determined. RESULTS: Melatonin ameliorated significantly the impairment of passive avoidance memory, the place navigation and spatial probe ability of mice induced by aluminum chloride. Melatonin was found to prevent significantly the decline of T-SOD, CuZn-SOD and GSH-Px activities, the increase of MDA content in the cortex and hippocampus of mouse brain induced by aluminum chloride. CONCLUSION: The results suggest that melatonin improves significantly the learning and memory impairment in mice induced by aluminum chloride, and this effect may be attributed to its antioxidation. PMID- 12567892 TI - [Apoptosis induced by diacetyldianhydrogalactitol and its mechanism in HL-60 leukemia cells]. AB - AIM: To investigate the apoptosis induced by diacetyldianhydrogalactitol (DADAG) and its mechanism in human HL-60 leukemia cells. METHODS: Inhibition of proliferation was measured by MTT assay. DADAG-induced apoptosis in HL-60 cells was observed by electron microscopy, flow cytometry and DNA fragmentation assay. The levels of Bcl-2 family proteins were detected by Western blotting. Caspase-3 activity was determined by ApoAlert CPP32 colorimetric assay kit. RESULTS: DADAG exhibited potent antiproliferative activity and induced apoptosis in HL-60 cells. After treatment with DADAG 8 micrograms.mL-1 for various times, the Bcl-XL protein level decreased in a time-dependent manner, while the Bad protein level was upregulated. The caspase-3 activity increased markedly after treatment with DADAG for 24 h. The apoptotic signals were suppressed by z-VAD.fmk (a general inhibitor of caspases), whereas z-DEVD.fmk, a selective inhibitor of caspase-3, only induced partial reversion of the apoptotic effects. CONCLUSION: DADAG induced apoptosis in HL-60 cells required caspase-3 activation and caspase-3 activation was related with Bcl-2 family members. PMID- 12567891 TI - [Effect of propylene glycol mannate sulfate on blood lipids and lipoprotein lipase in hyperlipidemic rat]. AB - AIM: To study the effect of propylene glycol mannate sulfate (PGMS) on blood lipids and lipoprotein lipase in hyperlipidemic rat, and its anti-hyperlipidemic mechanism. METHODS: PGMS was administered ig at different doses (37.8 mg.kg-1.d-1 and 75.6 mg.kg-1.d-1) to hyperlipidemic rats for three weeks and blood serum was obtained after starved 12 h. Total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were examined. The mRNA expression of lipoprotein lipase (LPL) in liver, spleen and artery was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: PGMS significantly decreased the levels of TC, TG and LDL-C and increased that of HDL-C in hyperlipidemic serum dose-dependently. PGMS was shown to increase the level of LPL mRNA expression, which is related directly to the controlling effects of PGMS on blood lipids. CONCLUSION: PGMS modulated blood lipids by promoting mRNA expression of LPL. This may be one important mechanism of PGMS to modulate blood lipids. PMID- 12567893 TI - [Antagonistic effect of hydroxysafflor yellow A on the platelet activating factor receptor]. AB - AIM: To observe the antagonistic effect of hydroxysafflor yellow A (HSYA) on the platelet activating factor (PAF). METHODS: Washed rabbit platelet (WRP) aggregation and rabbit polymorphonuclear leukocytes (PMNs) aggregation induced by PAF were observed by turbidimetric assay in vitro. The PAF receptor antagonistic effect of HSYA was investigated by radio ligand binding assay (RLBA). RESULTS: In RLBA the specific binding inhibition effect of HSYA was found to be concentration dependent in three different [3H]PAF concentrations. In the experiments, WRP aggregation and rabbit PMNs aggregation induced by PAF (9.55 x 10(-10), 9.55 x 10(-6) mol.L-1) were both inhibited by HSYA in a concentration-dependent manner in vitro. The IC50 of HSYA to inhibit WRP and rabbit PMNs aggregation was 0.99 and 0.70 mmol.L-1, respectively. CONCLUSION: The PAF receptor binding can be antagonized by HSYA. PMID- 12567894 TI - [Synthesis and insecticidal activity of the novel hydroxylbenzenedisulfonanilides compounds]. AB - AIM: To synthesize novel derivatives of hydroxylbenzenedisulfonanilide with high insecticidal activity against the Fasciola hepatica. METHODS: o-(m-, p-) Chlorphenol were used to synthesize the title compounds by sulphonation and nucleiphilic-substitution reaction. The uncoupling activity (insecticidal activity) of these compounds were tested by determining the influence on mitochondrial respiration control ratio (RCR) and the variation of inorganic phosphate in oxidative phosphorylation (delta Pi). RESULTS: Compounds 1-14 are new compounds. The structures of the compounds were determined by IR, HNMR and elemental analysis. Most new compounds have relatively high uncoupling activities, especially compounds 3, 5, 6 and 9. CONCLUSION: Compounds 3, 5, 6 and 9 will become novel fasciolicides and are worth further studying. PMID- 12567895 TI - [A new monoterpene glycoside from Paeonia lactiflora Pall]. AB - AIM: To study the chemical constituents of Paeonia lactiflora. METHODS: The constituents of P. lactiflora were separated by using various kinds of modern chromatography and was identified its structure on the basis of spectral analysis. RESULTS: A monoterpene glucoside named albiflorin R1 was isolated from the roots of Paeonia lactiflora Pall. In the structure of albiflorin R1, the aglycone connected with a glucose at its 2-OH while the hemiacetal hydroxyl in glucose moiety was free. CONCLUSION: Albiflorin R1 is a new monoterpene glycoside. PMID- 12567896 TI - [Study on chemical constituents of rhizome of Anemone raddeana]. AB - AIM: To investigate the chemical constituents of the rhizome of Anemone raddeana Regel, so as to find new active compounds. METHODS: The ethanol extracts of the rhizome of Anemone raddeana were obtained by silica column, HPLC. The structures of the compounds were elucidated by means of physico-chemical method and spectral analysis (IR, FAB-MS, 1HNMR, 13CNMR, DEPT, 1H-1H COSY, HMQC, HMBC). RESULTS: Nine compounds were isolated and identified as 27-hydroxyolean-12(13)-en-28-oic acid-3 0-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (1), eleutheroside K (2), Oleanolic acid-3-O-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D- glucopyranosyl (1-->4)]-alpha-L-arabinopyranoside (3), betulin (4), betulic acid (5), acetyloleamolic acid (6), evonymitol (7), oleamolic acid (8) and diosgenin (9). CONCLUSION: Compound 1 is a new compound, named raddeanoside 12. Compounds 3-7 were isolated from this plant for the first time. PMID- 12567897 TI - [Vectorial angle method for evaluating the similarity between two chromatographic fingerprints of Chinese herb]. AB - AIM: To evaluate the similarity between two chromatographic fingerprints automatically with computer. METHODS: Chromatogram can be treated as vector of hyperspace, and the similarity between them can be counted according to vectorial angle formula. This process was performed with software written in Visual Basic 6.0. The two main functions of this software are automatic peak tracking in two fingerprints under the same analytic condition and computing the similarity automatically. RESULTS: The HPLC fingerprints of eleven kinds of Evodia rutaecarpa (Juss.) Benth (a traditional Chinese herb) from different sources were obtained and the similarities were calculated with this software. This method was shown to be a good way to evaluate the similarity between two fingerprints. A sample washed seven times with hot water can be clearly discriminated from other samples of Evodia rutaecarpa (Juss.) Benth with similar results. CONCLUSION: This method is a good way to evaluate the similarity between two fingerprints and is helpful in quality control of traditional Chinese medicine. PMID- 12567898 TI - [Study on the mechanism of the interaction between montmorillonite and bacterium]. AB - AIM: To investigate the mechanism of the interaction between montmorillonite and bacteria by studying the reactions of different charges of montmorillonites with bacteria. METHODS: Bacteriostatic test: one loop of E. coli and Staphylococcus aureus at the concentration of 1 x 10(6).mL-1 was incubated to the plate culture medium containing different concentrations of montmorillonite, and incubated 24 h to observe the growth of bacteria. Bacterial adsorptive test: different amounts of montmorillonite were added into the artificially simulated intestinal solution (containing bacteria 1 x 10(7).mL-1). After the culture, the bacterial colonies were counted. RESULTS: The results showed that montmorillonite per se showed no bacteriostatic or bactericidal effect, but after exchange with metal ion and functional groups which inhibits bacteria, then it showed these activities. Adsorption was the main way between montmorillonite and bacteria. The special way of fixing bacteria into the "carriage" of montmorillonite gel which carry this structure was its pharmacological basis of curing diarrhea. The adsorption effect was related to layer charge density of the montmorillonites. CONCLUSION: Montmorillonite showed adsorption ability of bacteria with minus related to its layer charge, but has no bacteriostatic and bactericidal effect. PMID- 12567899 TI - [A fiber optic chemical sensor system for on-line monitoring the drug dissolution of rifampicin]. AB - AIM: To study a continuous, on-line and automatic monitoring method for dissolution of drugs. METHODS: A new fiber optical chemical sensor prepared by sol-gel immobilization of molecular probe was reported. Based on multiple quenching of flurescence, an instrument and software system was designed to on line monitor the drug dissolution of rifampicin. RESULTS: The concentration of rifampicin was linear with ln(F0/F) when concentration was in the range of 10-170 micrograms/mL-1. The relative coefficient was 0.9993. The procession of dissolution can be on-line monitored. Parameters were obtained directly from the dissolution curve. There is no distinctly difference when compared with the method of the Pharmacopoeia of People's Republic of China (P > 0.05). CONCLUSION: This procession analysis can reflect the real dissolution of drug and obtain the total information. PMID- 12567900 TI - [Preparation of diltiazem hydrochloride delayed-onset, sustained release tablet]. AB - AIM: To prepare dilitazem hydrochloride delayed-onset, sustained release tablet, which can not only provide the delay in release start, but also the constant release rate after a lag time. To analyze release mechanism and investigate the effect of outershell compositions on release behavior. METHODS: The delayed onset, sustained release tablets were prepared by dry-compression coated technology. The release profiles of uncoated cores and presscoated tablets were compared. Two parameters, time-lag (Tlag) and release rate (k), were used to evaluate the influence of factors, such as the amount of hydroxypropylmethylcellulose (HPMC) and poly-vinyl-pyrrolidinone (PVP) K30, the viscosity of ethylcellulose (EC) and HPMC, and the compression load on diltazam hydrochloride (DIL) release. Higuchi equation and Peppa's equation were used to analyze release mechanism. RESULTS: With the increase of HPMC amount or HPMC viscosity, Tlag was prolonged and k was decreased; With the increase of PVP K30 amount, Tlag was shortened and k was increased; EC viscosity and compression load above certain degree showed no effect on DIL release. CONCLUSION: The drug release from delayed-release tablet is controlled by erosion mechanism, Tlag is determined by the erosion rate of outer-shell. PMID- 12567901 TI - [Study on the characteristics of antisense oligodeoxy-neucleotides-liposomes complex and cellular uptake]. AB - AIM: To investigate factors affecting the properties of antisense oligodeoxy nucleotides (ASON)-liposomes complex and their cellular uptake. METHODS: Three types of blank liposomes were prepared by reverse-phase evaporation vesicles, and the complex were obtained through physical absorption. The light microscope was used to observe morphology characteristics of the complex. Drug loading capacity was analyzed by agarose gel electrophoresis. The transfected cell percentage and means fluorescence intensity were determined by flow cytometric analysis using M3 myeloma cell as a model. RESULTS: The neutral liposome showed no aggregation while the cationic liposomes appeared some different extent aggregation in different medium when associated antisense oligodeoxynucleotides. The drug loading capacity depended on the ratio of +/- and the cationic charge density on the lipid membrane. The two kinds of cationic liposomes appeared different principles of loading ASON. As far as cellular uptake, The neutral liposomes showed no improvement of cellular uptake of ASON. However, the cationic liposomes were shown to enhance the cellular uptake of ASON if the appropriate +/- charge ratio was used. The optimal cellular uptake was achieved when +/- charge ratio was at 0.5:1 and 1:1 for SA-I liposome and SA-II liposomes, respectively. CONCLUSION: The cationic liposomes improved the loading capacity and cell uptake of antisense oligodeoxynucleotides, which was determined by +/- charge ratio and charge density. PMID- 12567902 TI - [Preparation and in vitro release of tetramethylpyrazine phosphate pulsincap capsule controlled by an erodible plug]. AB - AIM: To develop a novel pulsatile drug delivery system of which the lag-time is controlled by an erodible plug (EP) and evaluate its release characteristics in vitro. METHODS: The impermeable capsule body was prepared by fulfilling method and the drug tablet and the erodible plug were made by wet granulating compression. Tetramethylpyrazine phosphate (TMPP) pulsincap capsule was prepared by sealing the drug tablet and fillers inside the impermeable capsule body with the EP. The influence factors on the lag-time such as the EP pharmaceutical properties and the dissolution condition were investigated by dissolution testing. RESULTS: Both the composition and the weight of EP influenced the lag time of the tetramethylpyrazine phosphate pulsincap capsule significantly. The lag-time prior to the drug release was enhanced when the content of gel-forming excipient (hydroxypropylmethylcellulose, HPMC) in the EP or the weight of EP was increased. The hardness of EP showed minor influence on the lag-time. In addition, the lag-time was shortened when the paddle speed was higher, while the pH value of the dissolution medium exhibited no significant influence on it. CONCLUSION: To meet the chronotherapeutic requirements, a pulsatile drug delivery system with a suitable lag-time can be achieved by adjusting the composition and the EP weight. PMID- 12567903 TI - [DNA sequencing and molecular identification of Patchouli and its substitute wrinkled gianthyssop]. AB - AIM: To analyze sequences of the nuclear ribosomal RNA small subunit (18S rRNA) gene and the chloroplast matK gene of crude drug Patchouli (Pogostemon cablin) in order to provide molecular evidence for identification of Patchouli drug. METHODS: To sequence the entire 18S rRNA gene and partial matK gene of Patchouli from Guangzhou and its substitute Wrinkled Gianthyssop (Agastache rugosa) from Sichuan using PCR direct sequencing and to detect the homology of two gene sequences between these two crude drugs. RESULTS: The complete 18S rRNA gene sequence is 1,805 bp in length for Patchouli from Guangzhou whereas 1,794 bp for Wrinkled Gianthyssop from Sichuan. The 3'-end sequence of matK gene is 521 bp (747-1,268 nt from upstream of matK gene) for these two crude drugs. Based on multiple sequence alignment, it is found that there are 18 variable sites and 11 aligned gap sites in 18S rRNA sequence, 49 variable sites in 3'-matK sequence between these two crude drugs. The homology is 98.4% for 18S rRNA and 90.6% for 3'-matK between two crude drugs, respectively. CONCLUSION: DNA sequencing can provide an accurate and reliable tool in the crude drug identification of Patchouli and its substitute Wrinkled Gianthyssop. PMID- 12567904 TI - [FK506-binding proteins and neurodegenerative diseases]. PMID- 12567905 TI - [Advances in search for acetylcholinesterase inhibitor based on the double active sites hypothesis]. PMID- 12567906 TI - [Effects of salvianolic acid B on beta-amyloid peptide neurotoxicity of primary cultured fetal rat cortical neurons]. AB - AIM: To study the roles of nitric oxide (NO) and nitric oxide synthase (NOS) on the glutamate (Glu) and beta-amyloid peptide [beta-AP (1-40)] mediated neurotoxicity in primary cultured fetal rat cortical neuron and the neuroprotective effects of salvianolic acid B (Sal B) against the beta-AP (1-40) and its mechanism of action. METHODS: With application of specific agonist and antagonist of NOS, establishment of the sodium nitroprusside (SNP), Glu and beta AP (1-40) neurotoxicity model, the cell viability, lactate dehydrogenase (LDH) efflux and NO release were detected by using morphological observation, MTT stain, spectrophotometric measurement and Griess method, respectively, in primary cultured fetal rat cortical neurons. RESULTS: Glu and beta-AP (1-40) were shown to increase the NO release of the neuron. Furthermore, nNOS was found to play an important role in the neurotoxicity of glutamate, iNOS may probably be involved in the neurotoxicity of beta-AP (1-40). Sal B (0.01, 0.10, 1.00 microgram.L(-1)) was shown to increase the cell viability, decrease the LDH release rate and inhibit NO release in a dose-dependent manner. CONCLUSION: These results suggest that the neurotoxicity of Glu and beta-AP(1-40) may be partly mediated through different types of NOS. Sal B was found to prevent the beta-AP(1-40) toxicity by directly or indirectly decreasing NO release. PMID- 12567907 TI - [Role of beta-adrenoceptor antagonists in the control of kindling models in rats]. AB - AIM: To study the role of beta-adrenoceptors (beta-AR) and their subtypes in the control of kindling models in rats. METHODS: Electrical amygdala kindling, pentylenetetrazol (PTZ) kindling models in rats and maximal electroshock (MES) in mice were used to observe the effects of beta-AR agonists and antagonists. RESULTS: The beta-AR antagonist propranolol 15 mg.kg-1 i.p. delayed the development of amygdala kindling; while propranolol 1-20 mg.kg-1 i.p. and beta-AR antagonist metoprolol 25-400 mg.kg-1 ig significantly increased the afterdischarge threshold and reduced the seizure severity in both kindling models (P < 0.01). beta 1-AR agonist dobutamine completely reversed the inhibitory effect of propranolol, while the beta 2-AR agonist terbutaline showed no effect on propranolol. Metoprolol 200 and 800 mg.kg-1 ig significantly inhibited MES in mice (P < 0.01). CONCLUSION: Central beta 1-AR is involved in the inhibitory effects on amygdala kindling and PTZ kindling models in rats. PMID- 12567908 TI - [Effects of baicalin on liver microsomal cytochrome P450 system]. AB - AIM: To investigate the effect of baicalin on liver microsomal cytochrome P450 system and the mechanism of liver protective action of baicalin. METHODS: Liver microsomal cytochrome P450, b5, aminopyrin N-demethylase (ADM), 7-ethoxycoumarin O-deethylase (ECD) and benzopyrene hydroxylase (AHH) activity were quantitated by UV chromatography. Activities of six cytochrome P450 isoforms were assayed with Western Blotting. RESULTS: Baicalin increased liver microsomal cytochrome. P450 level and ADM, ECD and AHH activity significantly. The three P450 isoforms, 1A1, 2B1 and 2C11, were also induced selectively by baicalin, but the b5 level, 3A2, 2D1 and 2E1 were not induced. CONCLUSION: Baicalin increases liver microsomal cytochrome P450 level and induces selectively 1A1, 2B1 and 2C11 of P450 isoforms in mice. PMID- 12567909 TI - [Effects of amiloride on calcium current in thyroxine-induced hypertrophied rat heart]. AB - AIM: The L-Type calcium currents were investigated in normal, hypertrophied and amiloride-treated rat ventricular myocytes so as to clarify the possible cause of the action potential lengthening that has been reported in thyroxine-induced hypertrophy. The interactions between agents such as thyroxine, Ang II and phenylephrine which induced hypertrophy with amiloride on the calcium current were analyzed. METHODS: Myocardial hypertrophy was induced by i.p. L-thyroxine in rats. The cell length, cell width and cell area were measured by image analysis technique. For recording ICaL, the whole cell patch clamp technique was used. Potassium currents were suppressed by replacing K+ ions with Cs+ ions in intracellular media, and sodium current was blocked by 50 mumol.L-1 tetrodotoxin. RESULTS: The Ca2+ current was found to be larger in hypertrophied cells (783pA) than in normal cells (433pA) and in amiloride-treated cells (429pA). However, no significant difference was observed in current density (6.54, 8.33, 5.74 pA/pF) in normal, hypertrophied and amiloride-treated cells. ICa displayed the same voltage dependence in three cell types. The potential giving 50% of activation (V1/2) moved towards more negative and the slope of activation curve was smaller in hypertrophied cells than in normal cells and amiloride cells. When expressed as percentages, the maximal increases in ICa were obtained with 1 mumol.L-1 Ang II and 100 mumol.L-1 phenylephrine in normal cells (+45.5% and +81.8%). Thyroxine 100 mumol.L-1 showed no effect on ICa. After giving amiloride, Ang II and phenylephrine did not increase the amplitude of ICa. CONCLUSION: At the dose of 0.5 mg.kg-1.d-1, amiloride (p.o.) was shown to prevent the amplitude of ICa from increasing and to be effective against increases of amplitude of ICa by Ang II and phenylephrine. PMID- 12567910 TI - [Neuroprotective effect of alpha-dihydroergocryptine depends on activation of nuclear factor kappa B]. AB - AIM: To investigate the relationship between the neuroprotective effect of alpha dihydroergocryptine (alpha-DHEC) and the activation of nuclear factor Kappa B (NF Kappa B). METHODS: Adult male rats were subjected to cerebral ischemia induced by middle cerebral artery occlusion (MCAO). DNA binding activity of NF-Kappa B was determined with electrophoretic mobility shift assay (EMSA) in animals subjected to varying durations of cerebral ischemia. Neuroapoptosis induced by ischemic damage was measured by deoxynucleotidy transferase-mediated dUTP nick end labeling (TUNEL) assay and flow cytometry (FCM) analysis. RESULTS: No change was observed in nuclear NF-Kappa B DNA binding in normal animal. A low level of constitutive NF-Kappa B DNA binding was detected in animals subjected to cerebral ischemia of 1 h, and significant increase in the amount of active NF-Kappa B in nuclear extracts was observed after cerebral ischemia of 3 h, 6 h, and 12 h. Peak of NF-Kappa B DNA binding was observed at the time point of 3 h. Animals subjected to cerebral ischemia of 3 h potentially initiates neuroapoptosis and activates NF-Kappa B in nuclear extract. Alpha-DHEC (100 micrograms.kg-1 and 150 micrograms.kg-1) showed significant protective effect on neuroapoptosis-induced by cerebral ischemia of 3 h, and inhibiting NF-Kappa B activation using 100 mg.kg 1 pyrrolidinedithiocarbamate (PDTC) in the continuous presence of alpha-DHEC, the neuroprotective effect of alpha-DHEC was blocked. CONCLUSION: The findings suggest that the neuroprotetive effect of alpha-DHEC may depend on the activation of NF-Kappa B. PMID- 12567911 TI - [Synthesis of droloxifene citrate and its new bioactivity]. AB - AIM: To investigate a feasible synthetic procedure of droloxifene and study on its new bioactivities. METHODS AND RESULTS: Droloxifene was synthesized using methoxybenzene and phenylacetic acid as starting materials, via Friedel-Crafts acylation, alkylation, demethylation, etherification, Grignard addition, elimination-dehydration, conversing configuration and forming citrate, totally 8 steps, overall yield 14.7% (9.2%). By pharmacological test, droloxifene citrate shows two new bioactivities: (1) obvious effect on reversing the MDR of K562/A02 cells and modulating mdrl, GST pi and TopoII alpha expression. (2) inducing apoptosis in cultured rat luteal cell. CONCLUSION: The improved synthetic procedure is shorter than the reported method by two steps and has advantages of simple separation, purification and conversion of configuration; easily available starting materials and reagents; concise operation. Two new bioactivities on reversing MDR and inducing apoptosis of luteal cell offered a clue in new drugs research and widened clinic application of droloxifene. PMID- 12567912 TI - [Structural elucidation of a new tetrahydroxystilbene of Radix Polygoni Multiflori Preparata and study on its cardiovascular activity]. AB - AIM: To investigate the cardiovascular active constituents of Radix Polygoni multiflori Preparata. METHODS: Compounds were isolated from the water soluble extract with column chromatography of Sephadex, ODS and HPLC. The compounds were identified on the basis of spectral analysis (IR, EI-MS, FAB-MS, 1HNMR, 13CNMR, 2D-NMR) and phytochemical properties. Its inhibitory effect on the proliferation of bovine vascular smooth muscle cells were bioassayed in vitro. RESULTS: One compound was isolated and identified as: 2,3,5,4'-tetrahydroxystilbene-2-O-(6"-O alpha-D- glucopyranosyl)-beta-D-glucopyranoside. CONCLUSION: Compound I is a new compound with cardiovascular activity. PMID- 12567913 TI - [Enantioselective pharmacokinetics of benproperine in healthy volunteers]. AB - AIM: To investigate the enantioselective pharmacokinetic process of benproperine in healthy volunteers. METHODS: An enantiospecific HPLC method was developed and used to determine the plasma concentrations of each enantiomer. Six healthy Chinese male volunteers received an oral dose of 60 mg (+/-)-benproperine. The ratios of the enantiomers in plasma samples were measured on a chiral AGP column. The plasma concentration of each enantiomer was then calculated using the ratios of enantiomers and total concentration of the two enantiomers previously measured. RESULTS: The plasma levels of (-)-(S)-benproperine were always significantly higher than those of its antipode in six volunteers. The mean AUC0 t and Cmax values for (-)-(S)-benproperine were 2.18 and 2.12 times higher than those of (+)-(R)-benproperine. There was no significant difference between the T1/2 for both enantiomers, tested by paired t test (P > 0.05). Half an hour after administration of benproperine, the S/R ratio in plasma samples was as high as 3.8, and in 2 hours it drastically decreased to about 2.2, then kept on till 24 hours. CONCLUSION: Benproperine showed significant enantioselective pharmacokinetics in the human after an oral dose of the racemate. PMID- 12567914 TI - [Effect of solution pH value on the chelation structure of zinc acexamate by electrospray ionization mass spectrometry]. AB - AIM: To study the effect of solution pH value on the chelation structure of zinc acexamate. METHODS: A series of samples at different solution pH values were prepared by 10% HCl or 1 mol.L-1 NH3.H2O. Then API/TOFMS with electrospray ion source was applied to assay the samples. The nitrogen curtain gas and nebulizer gas were adjusted to a constant flow rate of 0.6 microL.min-1 and 2 microL.min-1 respectively. Samples were infused into the electrospray interface using a 500 microL syringe pump at a flow rate of 5 microL.min-1. Mass spectra were acquired in positive ion modes by scanning over the range of m/z 100-1,000. RESULTS: The chelation structure of zinc acexamate is stable at pH 2.54 and it can be easy to form the ion (M + ZnY)+ (Y = CH3CONH(CH2)5COO-) and (2M + Na)+ in this condition. CONCLUSION: The drug is an effective antiulcer agent. It may decrease the acidity of stomach juice, and form a polymer to protect the ulcer. PMID- 12567915 TI - [Study on hydroxylated metabolites of benproperine in human urine]. AB - AIM: To investigate the hydroxylation process of benproperine in humans. METHODS: After an oral administration of 60 mg benproperine to ten healthy male volunteers, urine samples collected within 0-24 h were extracted by solid phase extraction and analyzed by liquid chromatography-ion trap mass spectrometry. A microbial transformation of benproperine combined with semi-preparative HPLC was used to get two reference substances of the hydroxylated metabolites, and their structures were then elucidated by NMR. Furthermore, the structures of conjugated metabolites were speculated based on their characteristics in MS fragmentation. RESULTS: Five hydroxylated metabolites of benproperine and some of their conjugates with endogenous glucuronic acid or sulfuric acid were found in urine of volunteers after the dose. The structures of two metabolites were identified as 4"-hydroxybenproperine and 4"'-hydroxybenproperine by comparison of HPLC retention times and mass spectra with those of authentics obtained from the microbial transformation. CONCLUSION: Hydroxylation of benproperine occurs preferentially at para-position of the alkoxyl group in the aromatic ring. The hydroxylated metabolites of benproperine in human urine mainly exist as their glucuronic acid or sulfuric acid conjugates. PMID- 12567916 TI - [Preparation of sterically stabilized liposomes containing all-trans retinoic acid]. AB - AIM: To prepare sterically stabilized liposomes containing all-trans retinoic acid, and to study its physicochemical properties. METHODS: The microstructure, the particle size and its distribution was determined by DSC, 1HNMR and 90 Plus Particle Size Analyzer and a TEM photo was taken. RESULTS: The size of conventional liposomes and sterically stabilized liposomes were about 0.35 and 0.42 micron. The sterically stabilized structure can be confirmed by TEM photo. CONCLUSION: The stericallly stabilized liposomes containing all-trans retinoic acid can be prepared by combination of film-casting and freeze-drying. PMID- 12567917 TI - [Interactions of insulin with dipalmitoylphosphatidylcholine liposomes]. AB - AIM: To study the interactions of insulin with dipalmitoylphosphatidylcholine liposomes. METHODS: The liposomes were prepared by reverse-phase evaporation vesicle method. The entrapped efficiency, size and distribution of the liposomes were determined, and the influences of insulin on entrapped efficiency, size and distribution of the liposomes were investigated. The influences of liposomes on the fluorescence emission spectra of insulin and the calcein leakage from the liposomes entrapped calcein induced by insulin were measured. RESULTS: Insulin has little influence on the size and distribution of the liposomes while the sizes of the liposomes were about 170-190 nm. The insertion of tyrosine of insulin into dipalmitoylphosphatidylcholine liposomes membrane was not deep. The insulin disturbed the liposomes membrane, induced the calcein leakage from the calcein-loaded liposomes. CONCLUSION: Amphiphilic, example insulin, may disturb the intact membrane of liposome through the interaction either hydrophobic or hydrophilic. The attention should be paid to the entrapment process of peptides into liposomes. PMID- 12567919 TI - [Advances in chemistry of norditerpenoid alkaloids]. PMID- 12567918 TI - [Dynamics of growth and total tanshinones accumulation in crown gall cultures of salvia miltiorrhiza]. AB - AIM: To determine the dynamics of growth and total tanshinones accumulation in crown gall cultures of Salvia miltiorrhiza in MS and 67-V liquid media. METHODS: Fresh, dry weight and total tanshinones yields in the cultures and in the medium were determined every 5 days in crown gall suspension cultures. RESULTS: In MS medium, the logarithmic growth phase of crown gall cultures in S. miltiorrhiza was from the 5th to 30th days, and the stationary growth phase was from the 30th to 35th days. From the 25th to 30th days, physiological activity of crown gall cultures was higher and their growth was better. However, in 67-V medium, the logarithmic growth phase of crown gall cultures was from the 10th to 25th days, and the stationary growth phase was from the 25th to 35th days. Total tanshinones were largely accumulated in the cultures and in the medium after 25 days. The total tanshinones yield (60 mg.L-1) was reached at the 35th day. CONCLUSION: Knowing the regularity of the growth and total tanshinones accumulation in crown gall cultures of S. miltiorrhiza will be helpful to take proper regulative measures in order to obtain the maximum total tanshinones yield. PMID- 12567920 TI - Pregnancy rhinitis and rhinitis medicamentosa. AB - PURPOSE: To present guidelines for the recognition, management, and referral of pregnancy rhinitis with a goal of improving the quality of the pregnancy experience for women afflicted with this condition. DATA SOURCES: A case study illustrating the presentation of a severe case of pregnancy rhinitis is followed a literature review of etiology, diagnosis, and management strategies. CONCLUSIONS: Pregnancy rhinitis is a condition of clinical importance that is frequently exacerbated by use of intranasal decongestant sprays. The resulting rhinitis medicamentosa exacerbates the nasal obstruction, with resulting sleep disruptions that negatively impact the experience of pregnancy. IMPLICATIONS FOR PRACTICE: Nurse practitioners may miss opportunities to provide support, anticipatory guidance, and symptom relief. Anticipatory guidance that stresses the critical necessity of avoiding nasal spray decongestants, environmental modification, use of intranasal saline, moderate exercise, and nasal strips for subjective relief may have the potential to markedly decrease escalation of the condition to a serious disorder. PMID- 12567921 TI - Screening strategies for group B streptococcus in the third trimester of pregnancy. AB - PURPOSE: To identify the best screening protocol to prevent neonatal group B streptococcal (GBS) sepsis through literature review. DATA SOURCES: Selected research articles, texts, and Internet sources. CONCLUSIONS: Centers for Disease Control and Prevention (CDC), American Academy of Pediatrics (AAP), American College of Obstetricians and Gynecologists (ACOG), and American College of Nurse Midwives (ACNM) have issued guidelines describing methods to identify pregnant women at risk of intrapartum transmission of GBS to their babies. Studies have been conducted to discover the superiority of one prevention method over the other but no consensus has been reached. IMPLICATIONS FOR PRACTICE: Before widely used prevention methods, approximately 8,000 babies each year became infected with GBS; of those, 400 died and many survivors suffered life-long sequelae. Adoption of an appropriate protocol can prevent transmission of GBS from a colonized mother to her infant. Clinicians should implement either universal culture-based or risk factor-based screening guidelines for prenatal diagnosis and intrapartum prophylaxis of GBS disease. PMID- 12567922 TI - Nurse practitioners' treatment of febrile infants in Utah: comparison to physician practice nationally. AB - PURPOSE: To determine nurse practitioners' (NPs') knowledge of published guidelines involving the treatment of infants with fever without a source and to compare the practice of NPs in Utah to physicians nationally. DATA SOURCES: Using a comparative descriptive design, a survey was sent to 450 NPs in Utah replicating the methodology of a 1998 survey of physicians. The NPs were asked about their evaluation decisions and management strategies for various ages of infants/toddlers with fever without a source. CONCLUSIONS: The practice decisions of NPs in Utah and physicians nationally were similar. Family NPs' clinical decisions aligned more closely with family practice medical doctors and pediatric NPs' with pediatricians. Overall, the younger the patient, the more likely were NPs to order labs and antibiotics or to hospitalize/refer to a physician. The fact that management decisions were not based upon existing protocols may account for the practice variation seen among NPs. IMPLICATIONS FOR PRACTICE: Management of a febrile infant is a common clinical scenario for the primary care provider. Although evidence-based protocols exist, the treatment of febrile infants remains controversial. Overall, NPs' evaluation and management of these infants were not shown to differ from physicians. Further studies are needed to determine why clinical guidelines are not being followed. PMID- 12567923 TI - Exploring the association between body weight, stigma of obesity, and health care avoidance. AB - PURPOSE: To explore the stigma of obesity and its effect on health care utilization, associations between self-esteem, attribution for weight, body mass index (BMI), satisfaction with medical care and the behavior of delaying/avoiding health care were examined. DATA SOURCES: A convenience sample of 216 women recruited from church sites in Las Vegas completed self-administered questionnaires. CONCLUSIONS: The findings show an increase in BMI is associated with an increase in the delay/avoidance of health care. Weight-related reasons for delaying/avoiding health care included having "gained weight since last health care visit," not wanting to "get weighted on the provider's scale," and knowing they would be told to "lose weight." IMPLICATIONS FOR PRACTICE: The obese are a stigmatized and vulnerable population. Nurse practitioners are challenged to be aware of attitudes towards obesity and to identify ways to promote continuity of care and regular health maintenance. The goals of Healthy People 2010 to reduce obesity-related morbidity cannot be met if health care is delayed/avoided. PMID- 12567924 TI - Prenatal care services in the public and private arena. AB - PURPOSE: This exploratory study described the prenatal care experience in the public and private arena from the perceptions of childbearing women using interpretive interactionism. DATA SOURCES: A face-to-face interview comprised of eight open-ended questions was used to obtain pregnant women's perceptions of their prenatal care experience and prenatal care needs. The purposive sample consisted of six women who received private prenatal care and 14 women who received public prenatal care. CONCLUSIONS: Five essential elements of the prenatal care experience were identified. Prenatal care was viewed as a cooperative effort between informal self-care and formal care by health professionals. Issues related to individuality and normality were important considerations in the delivery of prenatal care. IMPLICATIONS FOR PRACTICE: Controversy exists over the effectiveness of prenatal care in preventing poor outcomes, as the definition of what constitutes adequate prenatal care remains unclear. Advanced practice nurses (APNs) continue to play a pivotal role in the provision of prenatal care services. The expanded knowledge and skills possessed by APNs place them in a pivotal position to develop and implement individualized, developmentally appropriate prenatal care that the women in this study so desperately wanted. In addition, they can assist women in continuing the health promoting behaviors initiated prenatally through out their lifespan. PMID- 12567925 TI - Keeping an eye on a moving target: quality changes and challenges for nurses. PMID- 12567926 TI - Nurses, long-term care, and eldercare: impact on work performance. PMID- 12567927 TI - A multidisciplinary financial education research project. PMID- 12567929 TI - A lesson in patient safety from 'A beautiful mind'. PMID- 12567928 TI - Unlicensed assistive personnel retention and realistic job previews. AB - There is a rich literature addressing employee turnover and retention. Research demonstrates that realistic job previews increase employee retention by fostering development of realistic job expectations that match job and organization requirements. Realistic job previews use may be an effective strategy to decrease unlicensed assistive personnel turnover and increase retention.$ PMID- 12567930 TI - Responses to the nursing shortage: policy, press, pipeline, and perks. PMID- 12567931 TI - Clinic visit and waiting: patient education and satisfaction. AB - Teaching should be part of every patient's health care. Even though this study did not show that teaching improves patient satisfaction with the clinic visit, it did indicate that subjects who were taught by the nurse were more satisfied with the education received during that visit.$ PMID- 12567932 TI - A system approach to retention and recruitment. PMID- 12567933 TI - Messy conversations and the willingness to be disturbed. AB - We all long for a sense of community in our professional lives where we can bring our spirit, passion, and whole selves to work. When we work in environments of tribal warfare and clans carrying out subversive activities, we will choose to leave our souls at home to protect them from the toxicity of the work environment. For anyone who has been cared for by nurses who have left their soul at home, you have experienced the outcome of negative environments that cannot have effective conversations. Recently a nurse told about her experience of realizing one day that she had the right to be happy at work so that she could more effectively take care of her 5-year-old daughter when she returned home. When she saw this as a right, she made a decision to leave her organization and look for a place that valued conversations and harmony rather than aggression and lack of trust. The only way to improve the world is through relationships, and conversations are the prelude to creating that change. Having the courage to reach out and start the dialogue knowing full well that it might be very messy is the only way to end the fragmentation and loss of community we experience in our organizations. Wheatley (2002) describes the experience of "...sacred as a feeling that I belong here" (p. 133). In the process of conversations, we can explore and appreciate our differences but also feel more connected and one with each other as a result of having the courage to converse. When we see each other as equals, we also free ourselves to open up to new learning and relationships which can lead to excellence in clinical outcomes. It's all about the courage of conversations.$ PMID- 12567934 TI - Nursing informatics: an evolving specialty. AB - Health care organizations will continue to invest heavily in clinical information systems to help improve patient safety, reduce variability of care, and increase the efficiency of their staff. These clinical information systems can assist in reducing medication errors through sophisticated medication management solutions and provide clinical decision support to aid in the decision making of all clinicians. They can also help offset the effects of a growing worker shortage, especially hard-felt in nursing, by reducing redundant tasks, improving communication, and streamlining clinical data collection. As these clinical systems are implemented, there is an increased need for information management of all of this complex data. By bringing both clinical and information systems expertise to bear, the informatics nurse plays a crucial role in ensuring that these goals are met to help the organization maximize its use of clinical information technology.$ PMID- 12567935 TI - [Study on tissue culture of Poligonum multiflorum]. AB - In the present thesis, suitable medium for callus inducement and relationship between culture time and callus growth of Poligonum multiflorum were studied by mono-factorial and orthogonal design test. The experiments showed that the best medium for callus inducement was MS + 2,4-D 1 mg/L + 6-BA 1 mg/L + IBA 0.5 mg/L, dark culture was better than light culture, callus grew fast when it was cultured 12 days, whole plant could be induced by callus, the induce rate was 95%. PMID- 12567936 TI - [Chemical pattern recognition of traditional Chinese medicine kudingcha (I)]. AB - In this paper, the non-linear mapping method of pattern recognition was adopted to classify 78 samples of traditional Chinese medicine Kudingcha, with macro and trace elements as classified characteristic features. Ilex cornuta Lindl., Ilex latifolia Thunb. and Ligustrum lucidum Ait. were identified accurately. The results agree with those from pharmacognosy. This paper provides a new method for identification of traditional Chinese medicine. PMID- 12567937 TI - [Pharmacognostical identification of fructus glychrrhizae--an adulterant of fructus Xanthii]. AB - Pharmacognostic studies on the shape, microscopic structure and morphology of Fructus Glychrrhizae (Glycyrriza pallidifora)--an adulterant of Fructus Xanthii (Xanthium sibiricum) were carried out and compared with that of Frucus Xanthii to provide a basis for their identification. PMID- 12567938 TI - [Supercritical-CO2 fluid extraction of Allium stativum oils]. AB - Effective constituents from bulb of Allium stativum were extracted by supercritical-CO2 fluid. These constituents were analyzed by GC-MS. The results showed that oils from SFE-CO2 contained 12 components, two of them were first obtained from the plant. PMID- 12567939 TI - [Analysis of chemical components from Pheretima]. AB - The total fat, total nitrogens, free amino acids, inorganic elements of Pheretima have been determined. It will provide scientific evidence for the establishment of quality standards of pheretima. PMID- 12567940 TI - [The experiment research of ginger detoxification to Rhizoma Pinelliae]. AB - The experiment results have shown that Ginger can abate the stimulation of Phizoma Pinelliae on abdominal cavity of mice, reduce the torsion incidence rate of mice because of pain which is resulted to i.p. powder suspension of raw Rhizoma pinelliae. We also observed that Ginger can obviously inhibit the increase of capillary permeatility of abdominal cavity of mice, reduce the PGE2 content of inflammatory foot tissue of mice which is due to inject juice of fresh Pinellia Rhizoma. In addition, Ginger can increase the PGE2 content of mice gastric solution, antegonise the decrease of PGE2 content which is come from administration of raw Pinellia Rhizoma and protect the gastric mucous. Above mentioned results suggested that Ginger have detoxification in vivo. to toxin of Pinellia Rhizoma. PMID- 12567941 TI - [Oxygen free radical scavening activity and anti-lipid peroxidation of tea polyphenol]. AB - The oxygen free radical scavenging activity and anti-lipid peroxidation of tea polyshenol were studied in vitro. Tea polyphenol possessed significantly scavening effect on hydorxyl radical produced by Fenton recation and superoxide anion produced by xanthine-xanthine oxidase system (IC50 were 919.6 mg/L and 836 mg/L espectively). Tea polyphenol could significantly inhibit the lipid peroxidation of brain omogenate and cerbral mitochondrial membrane induced by hyderxyl radical. The results suggest that the anti-lipid peroxidation of tea polyphenol may be related to its scavenging effects on oxygen free radicals. PMID- 12567942 TI - [Effect of Limonium bicolor on bombyxmori life-span]. AB - The effect of limonium bicolor decoction (LBD) (1%, 5%, 10%) on the life-span of bombyxmori was investigated. It was found that LBD could obviously prolong not only average life-span and average of highest life-span of both female bombyxmori and moth, but also the daration of female silkworm stage. Furthermore, LBD clud be antianoxiasis and hunger-resistant, these results showed that BLD could promise longevity. PMID- 12567943 TI - [In vitro effects of ancient prescription Bazhengsan against Diplococus gonorrhoeae]. AB - The effects of three Chinese ancient prescriptions, Bazhengsan, Pixiefenqingyin and Wulinsan were determined in vitro against 4 strains of Diplocous gonorrhoeaes. The results showed antibacterial activity of Bazhengsan was the highest, with a MIC from 6.25% mg/ml to 25 mg/ml. when the concentration of Bazhengsan was 100 mg/ml or over, all the Diplocous gonorrhoeae tested would be killed within ten minutes. PMID- 12567944 TI - [Studies on control of root rot on Panax notoginseng]. AB - Chemical Control tests of pot, plot and field for Panax notoginseng root rot were conducted during 1995-1996. The results indicated that the chemical control is a effect measure to control rapidly occurring and spreading of Panax notoginseng root rot. It was the best treatment to coordinate use of bactericide and fungicide, obviously better than alone or mixed use of fungicide and also better than alone use of bactericide. In the pot and plot tests, the best coordinate treatment was the treatment of 10% phenazine plus 70% dexon plus 50% bavistin and plus water (1:1:500), the control effect was 70%; in the field test, the control effect of over 70% was also get with the treatment of 10% phenazine plus 70% dexon and plus small soil (1 Kg:1 Kg:150 Kg) per mu. PMID- 12567945 TI - [Chemical pattern recognition of traditional Chinese medicine kudingcha (II)]. AB - In this paper, the HPLC data from 78 samples of Kudingcha were treated with back propagation algorithm of artifical neural network pattern recognition, and the computer-aided classification of Ilex cornuta Lindl., Ilex latifolia Thunb. and Ligustrum lucidum Ait. was accomplished. This paper provides a scientific, advanced and feasible method for identification of traditional Chinese medicine. PMID- 12567946 TI - [Pharmacognostical identification on Chinese drug zhizhuliao]. AB - In this paper, 2 kinds of Zhizhuliao that original plants are Polygonum suffultum Maxim., P. amplexicaule D. Don. var. sinense Forb. et Hemal. are made an initial study on the characteristics, microscopical and physichemical sides. It provides scientific basis for its comprehensive development and ultilization, and to use in clinical. PMID- 12567947 TI - [Studies on the chemical constituents of Quercus mongolica]. AB - Seven compounds were isolated from Quercus mongolica Fisch., six (1-VI) of them were identified as fridelin (I), beta-amyrin (II), 1-octadecanol (III), beta sitosterol (IV), daucosterol (V) and gallic acid (VI) by means of chemical, physical and spectrosopic evidiences. PMID- 12567948 TI - [Analysis of amino acids in Trigonella foenumgraecum seeds]. AB - Seventeen amino acids in Trigonella foenumgraecum seeds are analyzed by means of HITACHI Model 835-50 High Speed Amino Acid Analyzer. Seven of them are essential amino acid for human body. The results provide references for evaluating the usefulness of T. foenumgraecum seeds in medicine and food. PMID- 12567949 TI - [Pharmacological studies on extracts from flesh of Cridtaria plecata and Hyriopsis cumingii]. AB - Adiminstration of extracts from flesh of Cridtaria plecata Lea and Hyriopsis cumingii Lea (EFCH), in 3.6 g/kg and 1.8 g/kg i.g for 7 days, can stimulate macrophage phagcytoic function and humoral immunolgical response to cook's RBC in mice. EFCH 3.6 g/kg i.g for 7 days, inhibite allergic reaction IV induced by 2,4 Dinitroclorobenzne. Swimming time of mice can be prolonged by EFCH 3.6 g/kg, 1.8 g/kg, 0.9 g/kg i.g and EFCH 3.6 g/kg enhence mice's toleranetowards hypoxia. Analgesic tests showed that EFCH 3.6 g/kg, 1.8 g/kg, 0.9 g/kg i.g have significent analgesic activities on mice. PMID- 12567950 TI - [The processing of Gekko gecko and the pharmacological action of its different parts]. AB - Both pharmacologic and toxic experiments are made on different parts of Gekko gecko Linnaeus. The results show that Gekko gecko Linnaeus' heads and its feet have obvious pharmacological action without any toxic or side effects, which provides a sound basis for the increase in its clinical utilization and the expansion of its medicinal parts as well as guarantee of safety and effectiveness after taking the medicine. PMID- 12567951 TI - [Comparison studies on pharmacodynamics of three kind of Carapax trionycis]. AB - Three kinds of Carapax Trionycis were used in rat to prevent and treat syndrom complex of dificiency of Yin. Compareing the improvement of symptoms of dificiency of Yin, it was found that all of them actted effectively in various degrees. The effect of ZhongHua Carapax Trionycis (ZHCT) was similar to those of Shan Rui Carapax Trionycis (SRCT), but the efficacy of Green Carapax Trionycis (GCT) was inferior to those of ZHCT and SRCT. All kinds of C. Trionycis could increase the function of responding to iriation in mice with deficiency of Yin. The intensity of ZHCT was similar to that of GCT, and superior to that of SRCT. They all could enhance phagocytic function of eticuloedothelial system obviously, promot the development of thymus in different degrees with no efect on spleen in young mice. The efficacy of increasing the immune function of ZHCT and GCT were more potent than that of SRCT. ZHCT and GCT significantly decreased the activity of SGPT in mice poisoned by CC14. ZHCT and GCT protected liver better than SRCT did. PMID- 12567952 TI - [Karyotype analysis on Iris lactea ssp. chinensis]. AB - In this paper, the chromosome number, karyotype and chromosome volume of Iris lactea ssp. chinensis were reported. The normal diploid was 2n = 32. Karyotype formula was based on Levan's publication in 1964, k (2n) = 32 = 22 m + 8 sm + 2T. According to the method of Kuo SR, the species chromosome based on the relative length was 2n = 32 = 6L + 4 M2 + 18 M1 + 4 S which belong to "2A" type according to the Stebbins' karyotype classification. The AS.K% was 62.5% from Arano's method. The total volume of chromosome groups was 34.95 micron 3. PMID- 12567953 TI - [Morphological and histological identification of Pholidota yunnanensis]. AB - The article reports the characters and microscopical characteristics of herba of Pholidota yunnanensis. The results can supply basis for the identification of the medicinal material. PMID- 12567954 TI - [Pharmacognostic identification of Chinese drug luxiancao]. AB - In this pper, the crude drugs of Balanophora involucrata and B. harlandii are identified on character, microproperties, UV spectra and TLC. Result of the identification may provide a basis for its diferentication, exploitage and utilization. PMID- 12567955 TI - [Studies on the chemical constituents of the root of Pimpinella thellungiana]. AB - Four compounds were isolated from the roots of Pimpinella thellungiana Wolff., their structures were identified as palmitic acid(I), 4-propenylphenol(II), pinoresinol(III), 2-methyl-2-hydroxy-5-methoxy berzo(d) hydrofuran-3-one(IV) by physicochemical constants and spectral analysis (UV, IR, MS, 1HNMR, 13CNMR, 13C 1HCOSY, DEPT), IV of them is a new compound, Pharmacological tests showed IV has a hypotensive effect. PMID- 12567956 TI - [A new triterpenoid from Adenophora polyantha]. AB - A needle crystal from the ethanol extract of Adenophora polyantha was isolated. Its structure was determined on the basis of its Elementary Analysis, IR, MS, 1HNMR, 13CNMR and chemical property and established as Isotaraxerone. PMID- 12567957 TI - [Study of anti-tumor action and influence on immune function of Allium stativum]. AB - Applying mice test on cancer model of S180 and Lewis cancer by using method clystered stomach(oral), observs the Anti-tumor function. Experiments showed that Allium sativum can inhibit tumor growth and Lengthen lift duration of mice, the rate of inhibiting tumor was 74.35% and life lengthening rate were 42.4%, with control grou comparison. There was profund difference between them. Besides, we discoved Allium sativum can raise immune function of mice. PMID- 12567958 TI - [Effect of aqueous extract of Lepidium apetalum on dog's left ventricular function]. AB - Left ventricular myocardial contractility, function of pump-blood and coronary flow in dog were increased by i.v. aqueous extract of Lepidium apetalum (AELA) 0.2 ml/kg (containing crude medicinal herb 2 g/ml). The effects were similar to i.v. isoperaline 10 micrograms/kg, but AELA did not obviously change heart rate, arterioveno-oxygen partial pressure difference and arteriovenooxygen solubility. The results indicated the AELA had advantages of cordial effect and increasing coronary flow instead of enhancing oxygen consumption of cardiac muscle. PMID- 12567959 TI - [Experimental study on the effect of abstinence with herbal preparation qingjunyin]. AB - The effect of herbal preparation Qingjunyin (QJY) on the animal model of drug dependence was studied. After the morphinistic models of white rats and mice were made by ever increasing doses of morphine each time, the animals were divided into large dosage QJY group, small dosage QJY group, sustained morphine group and control group. The withdrawal symptoms were observed after naloxone was given to the animals intraperitoneally. The results showed that QJY groups withdrawal symptoms of the addicted white rats were alleviated, the number of jumps of the addicted mice was reduced, the scores of the principal symptoms of the animals were decreased, while the body weights of the animals were gained, comparing with the control group, the difference was significant (P < 0.01). The effects of QJY enhanced in the large dosage group, which suggests QJY has abstinence effects. PMID- 12567960 TI - [Study on the inclusion of isodoniol-beta-cyclodextrin]. AB - The inclusion compound of isodoniol-beta-cyclodextrin was prepared by the liquidphase method in aqueous solution. And it was confirmed by thinlayer chromatography, differentical thermal analysis and specific rotation. The change (delta A) of ultraviolet spectrum was determined. The composition ratio of the inclusion compound between isodoniol and beta-CD was obtained by the continuous variation method. PMID- 12567961 TI - [Study on preparation of technological process of chaihuguizhi oral solution by the orthogonal design]. AB - The technological process of Chaihuguizhi Oral Solution was studied with the orthogonal design in four factors: the amount of collecting aromatic water, the time of decocting medicinal material, concentration of alcohol solution and the PH of oral solution. The best preparation conditions are selected. PMID- 12567962 TI - [Effect of various nutrient solutions on quality of Gastrodia elata]. AB - This paper dealed with the effect of media fertilization on the quality of Gastrodia eleta according to the results of tissue culture. The results showed that contents of nutrient elements, gastrodin and free amino acids in G. elata were affected by various nutrients solutions. The result of comprehensive analysis showed that the 1/2 MS medium was the fittest treatment of fertilization. PMID- 12567963 TI - [Identification on commercial drug jiangxiang from Hongkong]. AB - Accordint to macroscopical characters, histological structurs and analysis of essential oil, the comparison of drug Jiangxian from Hong Kong with Dalbergia sisso and Pterocarpus spp. was studied. The result showed that commercial drug Jiangxiang from Hong Kong is the heartwood of Pterocarpus masupium. PMID- 12567964 TI - [Pharmacognostical study of Chinese drug ningxia beimu]. AB - This paper deals with the macroscopic, microscopic and chemical characteristics of the bulbs of Fritillaria taipaiensis var. ningxiaensis. This crude drug could be distinguished by shape, size, hilum and striations of the sample grains. PMID- 12567965 TI - [Studies on the chemical constituents of Viscum liquidambaricolum]. AB - Two crystalline substances isolated from the whole plant of Viscum liquidambaricolum were identified as 5-hydroxy-7-methoxychromone and 5,7 dihydroxychromone. These compounds are first obtained from this genus. PMID- 12567966 TI - [Effects of hairy root huangqi on hemopoietic function]. AB - The effects of hairy root Huangqi on hemopoietic system were studied in this paper. The oral administration of 5-20 g/kg hairy root Huangqi per day for 5-12 days could increase the count of RBC and reticulocytes in mice with hemorrhagic anemia, raise the hemoglobin, RBC and hematocrit in mice with hemolytic anemia, enhance the number of WBC and nucleared cells in bone marrow of mice treated with cyclophosphamidum or x-rays. The experiments showed that hairy root Huangqi promoted the hemopoietic function. The effects were similer to those of natural Huangqi. PMID- 12567967 TI - [Pharmacologic studies on processed snakegall]. AB - The results showed that the processed snakegall could obviously relieve cough induced by the irritation of ammonia water in mice. It could also increase the secretion of respiratory tract and promote expectoration. PMID- 12567968 TI - [Study of the inclusion compound of rutin with beta-cyclodextrin]. AB - In order to improve the solubility of Rutin in aqueous solution and the stability, the inclusion compound of Rutin with beta-CD was manufacted by saturated aqueous solution method. The formation of the inclusion compound was confirmed by IR spectroscopy and differential scauning calorimetry. The content of Rutin in inclusion compound was determined by UV. The results indicated that the inclusion compound of Rutin with beta-CD was formed. Quantitative analysis demmonstrated that the molecular ratio of Rutin to beta-CD in the complex was 1:1 with the inclusion constant of 283.79 L/mol at 30 degrees C. The dissolubility of Rutin in water has added to 643.19 mg/L (20 degrees C). PMID- 12567969 TI - [Study on the extraction method and the opimun factor for Paeonia suffruticosa]. AB - The optimun extraction process were selected from three extraction methods according to the content of paeonol and it UV characters. Futhermore, the conditions of extraction process selected were studied with orthogonal test. The result showed that the yield of paeonol is 73.7%, RSD 2.93% (n = 3). PMID- 12567970 TI - [Study on GC fingerprint characteristic features of wuji baifeng pills]. AB - In this paper, the fingerprint characteristic features that can be used for identification of Wuji Baifeng Pills were obtained by gas chromatographic analysis. PMID- 12567971 TI - [Math-model of efficient cultivation of Angelica sinensis]. AB - Using two-time general rotation-constitution design, the math-mdele has been founded about Angelica sinensis prodution efficiency and quantity of nitrogen, phosphorus and potassium to use by doing experiments in many different spots and production demonstration for testing and verifying. Accordin to the modle, computer is used to simulate the better, through which people can get fresh Chinese angelica over 500 Kg per mu and also net income over 500 yuan per mu. It shows that this is the best combination. 95% density riliability for this plan is 6860-7600 plants, pure nitrogen 18.75-22.2 Kg, P2O5 9.1-11.1 Kg and K2O 4.46-5.37 Kg One mu. Proportion for nitrogen, phosphorus, potassium is 1:0.49:0.24. PMID- 12567972 TI - [Influence of seedling assortment on Panax notoginseng growth and yield]. AB - Making Panax notoginseng seedling assortment according to seedling size before transplanting, the result shows that the influence is better, the yield of root tuber and fruit is higher. Culturing good seedling is the fundamental measure to increase yield of P. notoginseng. PMID- 12567973 TI - [Comparison on the quality of scolopendra commodities from different gathering periods]. AB - Through physo-chemical analysis and pharmacological study to Scolopendra commodites, it showed that the quality of the commodities from autumn was better but gather, dry and store difficultly. According to market, gathering at Spring is available. PMID- 12567974 TI - [The analysis of donkey-hide glue, antler glue and tortoise shell glue by means of circular dichroism]. AB - Donkey-hide glue, antler glue, tortoise shell glue and their forgeries were identified by the analysis method of circular dichroism(CD) in this paper. The standard spectra and data were set up. The method is simple, fast and accurate. PMID- 12567975 TI - [Analysis of steroids in crude drugs by capillary gas chromatography]. AB - The steroids, cholest-4-ene-one and cholestane-3, 6-dion, were determined in 8 species of crude drugs of Haima and Hailong by capillary gas chromatography. PMID- 12567976 TI - [Studies on the chemical constituents of the bark of Betula platyphylla]. AB - The paper deals with the saperating of free phenolic hydroxyl dipolymer condensution-tannis B1, B2 and the compounds (+)-catechin, (-)-epicatechin from the bark of Betula platyphylla. Their structures have been evaluated by means of 1H, 13C-NMR technique and chemical ways. PMID- 12567977 TI - [Studies on the polysaccharide of Fomes marginatus]. AB - This paper reports that polysaccharose was prepared by ethanol precipitation of water extracted and by deproteinization with sevage method, purification obtain Fomes marginatus. The polysaccharose which was determined by using sephadex-G, PAGE, ACTME, TLC and PC is single composition. [alpha]D20 = 28.89, it content of polysaccharose is 22.62 percent. It was hydrolied of periodate oxidation, smith degradation and IR analysis, the results show that the main chain is composed of B-(1-->4), (1-->6) linkage. PMID- 12567978 TI - [Anti-injure effect of extract from orange (EOP) peel on germ cells of male mice]. AB - In present study, the anti-injure effect of EOP, on incidence of sperm abnormality and meiotic micronuclei of early spermatids in mice induced by cyclophosphamide was investigated. The results indicated that EOP itself was not a mutagen, but significantly inhibited increase of the rate of micronuclei and the sperm abnormality by CP. The rates of sperm abnormality and meiotic micronuclei of early spermatide of EOP before CP using group were lower obviously than those of CP could before EOP using group (P < 0.01). The results suggested that the CP could permeate blood-testis barrier into germ cells of male mice and induced mutation on genetic material. PMID- 12567979 TI - [The immune activity of polysacharide of chuanniuxi]. AB - It reported the immune activity of polysacharide of Chuanniuxi to mouce's RES, PFC, lymphocyte transformation and NK Cell. The results show that the polysacharide can improve the activity of RES, PFC and NK Cell, but can't increace the lymphocgte stimulation index. PMID- 12567980 TI - [Identification of the binding site on ICAM-1 for red blood cells infected by Plasmodium falciparum]. AB - OBJECTIVE: To identify the binding site on ICAM-1 to PRBCs in order to explore anti-adhesive agent against cerebral malaria. METHODS: Monoclonal antibody 15.2 against ICAM-1 domain 1 was chosen as target molecule to screen mimetic peptides of ICAM-1 from a 12-mer random peptide library. Three rounds of biopanning were carried out and then ELISA, competitive ELISA, dot-ELISA and Western blotting were used to evaluate the binding character between phage-borne peptides and McAb 15.2. The insert DNA sequences of positive clones were determined and their amino acid sequences were deduced. RESULTS: Thirty clones from the third round were randomly selected, and 26 of them were found positive by sandwich ELISA. The competitive ELISA test proved that most phage-borne peptides could competitively inhibit the binding of antibody (15.2 McAb) with ICAM-1. Analysis of DNA and amino acid sequences indicated that over a half positive phage clones expressed 12-mer peptide KLYLIAEGSVAA. Comparison of peptide K(XX) L(XXX) GSV with the 64 73 aa of primary sequence of ICAM-1 showed a 50% homogeneity. CONCLUSION: These peptides displayed by phage may be analogs of ICAM-1, K..L...GSV probably plays a significant role on the binding reaction of ICAM-1 and PRBCs. PMID- 12567981 TI - [Epidemiological survey of hydatid disease in Hualong County, Qinghai Province]. PMID- 12567983 TI - [Preliminary observation on Pneumocystis carinii infection in lung cancer patients]. PMID- 12567982 TI - [Preliminary observation on specific antibody level and reduction of ovulation induced by recombinant Schistosoma japonicum 26 kDa GST antigen in water buffaloes]. AB - OBJECTIVE: To observe the specific antibody level and reduction of egg-laying induced by a recombinant Schistosoma japonicum 26 kDa GST antigen (rSjc26 GST) in water buffaloes. METHODS: 20 water buffaloes were randomly divided into two groups, the vaccination group and control group, with 10 buffaloes each. The subjects in vaccination group were immunized with rSjc26 GST antigen while the control received adjuvant only. After challenged with S. japonicum cercariae, the anti-rSjc26 GST antibody level and the numbers of eggs and miracidia in stool were detected. RESULTS: The anti-rSjc26 GST antibody appeared 1 month after immunization with rSjc26 GST antigen and maintained a high level for 12 months. Numbers of eggs (EPG) and miracidia (MPG) in vaccination group were significantly lower than those in control group during the period of day 50 to day 90 post challenge. However, EPG and MPG tended to decrease starting from day 100 post challenge in both groups. The difference of EPG and MPG between the two groups diminished progressively, and both groups showed zero egg count from day 330 on post challenge. CONCLUSION: The specific anti-rSjc26 GST antibody was detected in vaccinated water buffaloes and maintained a high level for 12 months. The vaccination showed a significant effect to the reduction of ovulation in the first three months after S. japonicum cercariae challenge. PMID- 12567984 TI - Proliferation and cytokine production of lymphocytes from Clonorchis sinensis infected rats in response to stimulators in vitro. AB - OBJECTIVE: To study the rat lymphocyte proliferation, differentiation and cytokine production in response to Clonorchis sinensis infection. METHODS: The lymphocyte proliferation and cytokine production (IFN-gamma, IL-2, IL-4, IL-10) in response to mitogen phytohaemagglutinin (PHA), C. sinensis excretory-secretory antigen (ES Ag), C. sinensis crude antigen (crude Ag) and Anisakis larvae antigen were detected in vitro from splenic lymphocytes (SLC) and mesenteric lymph node cells (MLNC) of rats infected with C. sinensis. Statistical analysis was performed by Sigma Plot System. RESULTS: Lymphocyte proliferations in MLNC were higher than that in SLC. At concentrations of 3 x 10(6) or 9 x 10(6) cells/well, lymphocyte proliferations were significantly higher in both SLS and MLNC than in the control with cell alone (P < 0.01). At the lymphocyte concentration of 5 x 10(6) cells/well and stimulator concentration of 5 or 10 micrograms/ml, significant lymphocyte activation was observed. Under the same culture condition (5 x 10(6) cells/ well with 10 micrograms/ml stimulator), cytokine IFN-gamma and IL-10 production in vitro increased significantly in MLNC. CONCLUSION: Concentrations of 5 x 10(6) lymphocytes/well and 10 micrograms/ml stimulator were selected as the optimal culture condition for activation of lymphocyte proliferation and cytokine production. Since the production of Th1-type cytokine IFN-gamma and Th2-type cytokine IL-10 was much enhanced from MLNC of C. sinensis infected rats, it is considered that C. sinensis ES Ag may stimulate lymphocyte proliferation and cytokine production in C. sinensis-infected rats in vivo. PMID- 12567985 TI - [Observation on the survival and invasibility of Ascaris suum egg preserved at low temperature for 5 years]. PMID- 12567986 TI - [Purification and immunogenicity identification of the recombinant protein related with specific IgE against Schistosoma japonicum]. AB - OBJECTIVE: To purify the specific IgE antibody-related recombinant protein of Schistosoma japonicum and to identify its immunogenicity. METHODS: The recombinant plasmid Sj43B/pGEX-6p-1 was expressed in E. coli BL 21. The inclusion body of the fusion protein was washed by TNMFX buffer and separated by FPLC. After renaturation, the fusion protein was used to vaccinate the mice. The specific IgG and IgE antibodies were detected by dot-ELISA and Western blotting analysis, respectively. RESULTS: Most of the proteins mixed with the inclusion body of the recombinant protein could be eliminated by washing with TNMFX buffer. The purified recombinant fusion protein could be obtained by FPLC separation. The experiment on mice immunized with the fusion protein showed that the specific IgE antibody was generated against the target part of the fusion protein, but not the specific IgG antibody. CONCLUSION: The fusion protein expressed by the recombinant plasmid Sj43B/pGEX-6p-1 could induce specific IgE response of the immunized mice. PMID- 12567987 TI - [Investigation of Paragonimus westermani infection in juvenile crabs in pargonimiasis endemic area in Jing'an County, Jiangxi Province]. PMID- 12567988 TI - [Application of multifactor spatial composite model to predict transmission tendency of malaria at national level]. AB - OBJECTIVE: To predict the transmission tendency of malaria at national level by application of geographic information system(GIS) technique. METHODS: With the assistance of ArcView 3.0 a software and its spatial analyst extension, the surface spatial analysis on three natural factors, namely, total growing degree days(TGDD), precipitation and relative humidity, were conducted individually. The map calculation was preformed based on the three factors' ratio of 5:3:2 resulted from the Delphi investigation. RESULTS: The individual maps and composition map of TGDD, precipitation and relative humidity were created, respectively, based on the spatial composite model, which were used to predict the transmission tendency of malaria at national level. CONCLUSION: The high risk areas for malaria transmission, predicted by the spatial composite model based on the multilayers of environmental factors, are correlated with the previous reports. This will, therefore, provide information for predicting malaria transmission by multiple factors in a larger area. PMID- 12567989 TI - [Reexamination of specific antibodies in sera of cystic echinococccosis patients with IgG negative seroresponse]. AB - OBJECTIVE: To explore the factors of false negative antibody response in patients with echinococcosis granulosus (Eg) for improving immunodiagnosis. METHODS: Indirect ELISA and sandwich ELISA were used to detect the specific antibody of IgG subclass (IgG1, IgG2, IgG3, IgG4) and IgA, IgM, IgE, as well as circulating antigen (CAg) and immunocomplex (CIC) in sera of Eg patients with negative response of total IgG. RESULTS: Among 42 sera with IgG negative seroresponse, 32 were positive with IgG subclass, IgA, IgM or IgE antibody, 10 were negative in all 7 kinds of antibody response. The detection rate of specific IgG1, IgG4, IgA, IgM and IgE was 42.9%, 11.9%, 28.6%, 26.2% and 21.4% respectively, being significantly higher than in sera of the control. IgM level in children was higher than that in adults. IgG subclass in patients with liver Eg was higher than that of pulmonary hydatidosis, when testing with IgG1 combined one or two of the other six Ig antibodies. The highest positive rate (64.3%) was seen in IgG1 + IgA + IgM antibody system. The positive rate of CAg and CIC in IgG negative patients was 28.57% and 30.95%, respectively. CONCLUSION: The factors involved with seronegative response of total IgG in Eg patients might be low specific IgG, variant Ig antibody expression and formation of CIC. Combined detection of IgG1 + IgA + IgM could enhance the sensitivity of serological tests in Eg patients. PMID- 12567990 TI - [Report on a case of Dioctophyma renale infection]. PMID- 12567992 TI - [Improvement in the preparation of Paragonimus westermani chromosome for karyotype analysis]. AB - OBJECTIVE: To improve Paragonimus westermani chromosome preparation technique. METHODS: After being exposed to colchicine, the gonadal cells of P. westermani were treated by the following procedures: hypotonicity, fixation, dropping onto a slide and staining. RESULTS: The chromosome number of P. westermani is 22, and the karyotype is 2m + 6Sm + 14t chromosome. CONCLUSION: The improved technique of chromosome preparation is feasible to operate and the chromosome is clear enough for karyotype analysis. PMID- 12567991 TI - [Construction and expression of single chain Fv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum]. AB - OBJECTIVE: To construct single chain Fv (scFv) gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum. METHODS: The heavy and light chain variable region genes of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum were inserted into two corresponding sites of expression vector pTHA90, and a scFv gene was constructed with a short peptide (Gly4Ser)3 linker gene. The recombinants were determined by digesting with XhoI/SpeI, XbaI/EcoRI and XhoI/EcoRI, and then were introduced into E. coli Top10. The antigen binding activity of expressed product was detected with ELISA. RESULTS: The recombinants were determined by digesting with endonucleases and expected bands were identified. The value of expressed scFv was 3 times higher than negative control by ELISA(OD492 = 1.06). CONCLUSION: The scFv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum was successfully cloned, and the expressed scFv fragment could interact specifically with antigen NP48. PMID- 12567993 TI - [Therapeutic efficancy of tradition Chinese medicine "nang chong san" on derme muscular cysticercosis by local injection]. PMID- 12567994 TI - [Expression and characterization of 21.7 kDa membrane protein of Schistosoma japonicum]. AB - OBJECTIVE: To express, purify and characterize the 21.7 kDa membrane protein of Chinese strain S. japonicum (SjC21.7). METHODS: The gene of SjC21.7 was subcloned into the expression vector pGEX-4T-3 to form recombinant plasmid. The recombinant plasmids were transformed into E. coli BL21 and the GST-SjC21.7 fusion protein was expressed by IPTG induction. The recombinant SjC21.7 molecule was prepared by affinity chromatography and digested by thrombin. The Kunming strain mice were immunized with the recombinant SjC21.7 molecule to produce anti-SjC21.7 antibody. The purified SjC21.7 was recognized by the immunized mouse serum and the sera of rabbits infected by S. japonicum. RESULTS: The SjC21.7 gene was subcloned into expression vector pGEX-4T-3, then transformed into E. coli BL21 to express the GST-SjC21.7 fusion protein. The recombinant SjC21.7 molecule obtained from the fusion protein could stimulate the mice to produce a high titer of specific antibody and could be recognized by sera of both the immunized and infected rabbits. The sera of immunized mice could also recognize the 21.7 kDa protein molecule of the adult worm antigen (AWA). CONCLUSION: The recombinant and purified SjC21.7 was prepared and showed similar immunological characteristics to the natural SjC21.7 molecule, providing a basis for further investigation of the immunological protection of the recombinant SjC21.7. PMID- 12567995 TI - [Cloning and sequencing of the gp23 gene encoding a surface antigen on sporozoites of Cryptosporidium parvum]. AB - OBJECTIVE: To clone and sequence the gp23 gene encoding a surface antigen on the sporozoites of Cryptosporidium parvum. METHODS: Genomic DNA was isolated from oocysts of Cryptosporidium parvum. The gp23 gene was amplified by polymerase chain reaction (PCR) and cloned into pMD18-T vector and sequenced by the methods of dideoxy-mediated chain termination. RESULTS AND CONCLUSION: The gp23 gene was 345 bp in length and included an open reading frame encoding a protein of 114 amino acid residues. The homology of the nucleotide and amino sequences of the gp23 gene was 97.3% and 98.2% to that in the GenBank, respectively. The gp23 gene cloned contained 6 nucleotides more than that in the GenBank. PMID- 12567996 TI - [Identification of epitope of 22.6 kDa antigen of Schistosoma japonicum with phage-displayed random peptide library]. AB - OBJECTIVE: To identify the epitopes of 22.6 kDa antigen of Schistosoma japonicum (Sj22.6). METHODS: A 12-mer library displayed on pIII of fd phage was used to screen the epitopes of Sj22.6 with the purified multiple clone IgG antibody against the Sj22.6 antigen. Five rounds of biopanning were carried out and fourteen clones from the fifth round biopanning were randomly selected and identified by Western blotting. The mice were immunized with the obtained positive clones and the antibody titers of sera from the mice were detected. The clones which could stimulate the mice to produce anti-Sj22.6 antibodies were sequenced and their amino acid sequences were compared with that of Sj22.6. RESULTS: Six clones selected from the fourteen ones could stimulate the mice to produce anti-Sj22.6 antibodies analysed by Western blotting. The amino acid sequence of one epitope showed high homology with Sj22.6. CONCLUSION: Four epitopes of Sj22.6 were obtained. One may be a structure epitope and the other three were mimic epitopes. PMID- 12567997 TI - [Application of ultrasound scan during surgery of cystic hydatidosis with polylamination and multiple daughters cysts]. PMID- 12567998 TI - [Molecular variation based on mtDNA-COII gene of members of the Anopheles minimus group]. AB - OBJECTIVE: To investigate intra- and inter-species molecular variations and phylogeny of the five members of the Minimus Group of Anopheles subgenus Cellia in China: An. aconitus, An. varuna, An. jeyporiensis, An. minimus A and An. minimus C, and to report the DNA sequence divergence among these species at a mitochondrial locus (cytochrome oxidase II). METHODS: Single mosquito's legs were digested to extract DNA. COII gene was amplified, sequenced and analysed; and the phylogenetic tree of members of An. minimus group was reconstructed by maximum likelihood method (DNAML). RESULTS AND CONCLUSION: The data confirmed the presence of two cryptic species, A and C, within An. minimus complex in the research localities; and two species differed by 2.3% in the COII sequences owing to 16 nucleotide substitutions. There was less variation between the two species than other members of the Minimus Group. PMID- 12567999 TI - [Changes in element content in Aedes albopictus infected by dengue virus type II]. AB - OBJECTIVE: To study the changes in the content of elements in Aedes albopictus after dengue virus type II (DV2) infection. METHODS: DV2 were injected into the thorax of Ae. albopictus. The content of elements in Ae. albopictus was determined on days 5, 10 and 20 after infection respectively and compared with those in the same instar non-infected mosquitoes. RESULTS AND CONCLUSION: For the group of day 5 after DV2 infection, the content of elements Ca, S and Cr increased remarkably compared to those in the control group; while the content of elements Zn, Mo, Pb, Ce and Cl reduced remarkably. For the group of day 10 after infection, the content of Na, Mg, Zn, Se, Ce and S increased remarkably compared to those in the control; only that of Fe was lower remarkably than the control. For the group of day 20 after infection, the content of elements Ti and Pb increased remarkably than that of the control; while the content of Na, Zn, Cr and Ce reduced remarkably. PMID- 12568000 TI - [Clinical observation of 13 cases of paragonimiasis complicated with bronchial asthma]. PMID- 12568001 TI - [Target chemotherapy of intestinal nematode infection in area with low endemicity]. AB - OBJECTIVE: To evaluate the control measures for intestinal nematodiasis in endemic area with low prevalence and intensity of infection. METHODS: Target chemotherapy was carried out in high-risk population based on the epidemiological characteristics such as age and clinical findings. Albendazole and mebendazole were administered each 200 mg once daily every year for 3 or 5 years. Saturated brine floatation and Kato-Katz thick smear techniques were used for stool examination to evaluate the efficacy of treatment. RESULTS: Two hundred residents from each of the three investigation villages were selected for target chemotherapy once a year for three years. The prevalence of intestinal nematodes decreased from 6.2% in 1995 to 5.4% in 1996 and 3.2% in 1997, and remained at 2.3% after three years in 2000. One control village where only primary school students were treated once a year for 5 years, the prevalence of Ascaris and Trichuris infection also decreased from 1.4% and 4.2% in 1995 to 0.9% and 1.4% in 2000, respectively. The target chemotherapy on the predisposed population to hookworm infection showed that the prevalence in the population above 41 years old was declined from 19.4% to 10.9%. CONCLUSION: The target chemotherapy is an economical and effective approach for the control of intestinal nematode infection in endemic area with low prevalence and intensity of infection. PMID- 12568002 TI - [Endoscopic diagnosis on hookworm infection in 30 cases]. PMID- 12568003 TI - [Investigation of inpatient cases of food-borne parasitic encephalopathy]. AB - OBJECTIVE: To investigate the clinical features of the patients with encephalopathy caused by food-borne parasites. METHODS: Questionnairing was carried out to collect and analyze clinical data of cerebral form of food-borne parasitic diseases in the hospital during the past five years. RESULTS: Among 190 discharged medical histories, 115 cases were valid for investigation, the number of males was 73, females 42, with a ratio of 1.74:1. Among these patients, 20.9% (24/115) had a history of eating raw meat. For discharge diagnosis, neurocysticercosis accounted for 92.2% (106/115), cerebral paragonimiasis 3.5% (4/115), sparganosis 2.6% (3/115), and angiostrongyliasis cantonensis and gnathostomiasis 0.9% (1/115) each. 13.9% (16/115) of the patients were hospitalized for three times or more. CONCLUSION: More attention should be paid to food-borne parasitic encephalopathy. PMID- 12568004 TI - [Therapeutic effect of dihydroartemisinin combined with naphthoquine phosphate in patients with falciparum malaria]. AB - OBJECTIVE: To observe the therapeutic efficacy of dihydroartemisinin combined with naphthoquine phosphate in patients with falciparum malaria. METHODS: Patients with Plasmodium falciparum were selected as the subjects, treated with a single dose of dihydroarteminisinin 160 mg combined with naphthoquine phosphate 400 mg (for adults) and followed up in preselective time by blood and temperature examination for 28 days after drug administration. RESULTS: 37 patients with falciparum malaria were treated and followed up. One patient had recrudescence and the cure rate in 28 days was 97.3%(36/37). The mean fever clearance time was (15.8 +/- 8.7) hours; the mean parasite clearance time was (27.6 +/- 13.2) hours; the mean reduction parasite rate in 24 hours was 96.7% +/- 26.5%. No apparent side effect was observed. CONCLUSION: A combination of dihydroartemisinin and naphthoquine is effective for the treatment of patients with falciparum malaria. PMID- 12568005 TI - [Relationship of Demodex infection of facial skin disease]. PMID- 12568006 TI - [Current status and prospects of the research on sex-specific gene of schistosomes]. PMID- 12568007 TI - [Clinical analysis of 142 cases of cerebral cysticercosis]. PMID- 12568008 TI - [DNA sequencing of the AChE fragment of Aedes albopictus]. PMID- 12568009 TI - [Surveillance after the interruption of malaria transmission in Loudi City]. PMID- 12568010 TI - [Modification and optimization of agricultural structure and schistosomiasis control]. PMID- 12568011 TI - [Sequence analysis of MESA gene of Plasmodium falciparium FCC1/HN strain]. PMID- 12568012 TI - [Investigation of intestinal nematode infection in Jiande City, Zhejiang Province]. PMID- 12568013 TI - [A case of unfinequently seen toxoplasmosis]. PMID- 12568014 TI - [Study on treatment of multi-drug resistant falciparum malaria by using a combination of dihydroartemisinin and pyronaridine]. AB - OBJECTIVE: To provide a combined medication scheme for the treatment of multi drug resistant falciparum malaria. METHODS: Combined administration of dihydroartemisinin and pyronaridine was given to the 32 cases of falciparum malaria cases with multi-drug resistance. The indices for evaluation on day 14, 21, and 28 after treatment included the mean fever subsidence time, mean asexual form clearance time, mean recrudescence time of asexual form and recrudescence rate, proportion of gametocyte carriers, mean density of gametocytes and its mean clearance time, cure rate and rate of side-effects. A double blind clinical test was performed with standard schemes of dihydroartemisinin (20 cases) and pyronaridine (25 cases) as control. RESULTS: The mean fever subsidence time of treated patients by dihydroartemisinin/pyronaridine combination, dihydroartemisinin and pyronaridine was 35.7 +/- 24.7 h, 52.6 +/- 38.9 h and 35.8 +/- 16.5 h respectively, showing a significant difference between the combination group and dihydroartemisinin groups (P < 0.01). The mean asexual form clearance time was 23.8 +/- 10.1 h, 22.9 +/- 6.5 h and 49.4 +/- 20.3 h respectively, showing significantly faster in the combination group than the pyronaridine group (P < 0.01). The recrudescence rate was 0, 4.2% and 0 respectively. The proportion of gametocyte carriers was 20.0%, 16.7% and 60.9% respectively, with a significantly higher rate in the group of pyronaridine than the group of combination (P < 0.01). CONCLUSION: The combination of dihydroartemsinin and pyronaridine is an ideal medication scheme for the treatment of falciparum malaria cases with multi-drug resistance. PMID- 12568015 TI - [Preparation and staining of Domodex folliculorum specimen]. PMID- 12568016 TI - [Cloning and sequence analysis of LACK gene of Leishmania donovani isolates from plain and desert foci of China]. AB - OBJECTIVE: To determine the nucleotide sequence of the LACK (Leishmania homologue of receptors for activated protein kinase C) gene of Leishmania donovani isolates from plain foci (L. d SD1) and desert foci (L. d XJ771) of China, and to find out the difference of the sequence of LACK gene with other Leishimania spp. and also the isolate from hill foci of China. METHODS: The LACK genes of L. d SD1 and L. d XJ771 were amplified by RT-PCR and cloned into pUC18 vector respectively, sequenced by the dideoxy chain termination method, then analyzed and compared with that of other isolates. RESULTS: The LACK genes of the two isolates were successfully cloned. Both of the 2 fragments were 942 bp in length. Comparison of the two nucleotide sequences with that of other Leishmania spp. in GenBank showed that the identities were more than 97%, and the identities of the nucleotide sequences of LACK genes of the three L. d isolates from plain, desert and hill foci of China were more than 95%. CONCLUSION: High identities exist among the nucleotide sequences of LACK genes of the three L. d isolates from the three foci of China. PMID- 12568017 TI - [One case of malaria with repeated attacks]. PMID- 12568018 TI - [Protective immunity induced by the nucleic acid vaccine of SjC 21.7 in mice]. AB - OBJECTIVE: To observe the protective immunity induced by the nucleic acid vaccine of 21.7 kDa membrane protein molecule of Schistosoma japonicum Chinese mainland strain (SjC 21.7) in BALB/c mice. METHODS: A pair of primers (P1 and P2) was synthesized according to the DNA sequence of the SjC21.7. The ORF sequence of SjC21.7 was amplified by PCR, and the Kozark sequence was added to the position of initiator. The gene fragment was inserted into the eukaryotic expression plasmid pcDNA3.1 to form the recombinant plasmid SjC21.7-pcDNA3.1. Forty-eight BALB/c mice were divided into three groups: control, test and boost. Each mouse was injected in quadriceps femoris with plasmid pcDNA3.1 (control) or recombinant plasmid SjC21.7-pcDNA3.1 (test, boost); for the boost group, with additional P35 pcDNA3.1 and P40-pcDNA3.1. All mice were immunized three times with an interval of 2 weeks, challenged each with 45 cercariae of S. japonicum at the 30th day after final immunization. At day 45 after challenge, all mice were sacrificed, the numbers of worms and hepatic eggs were counted. Antibody level in the sera of mice before and two weeks after immunization was determined with ELISA. The expression of the target gene in quadriceps femoris was observed with immunohistochemistry. RESULTS: The immunohistochemistry analysis showed that there were specific antigens expressed in the local tissue of the test group mice. There was specific IgG in the serum of partial mice in test and boost groups. Compared with the control group, the worm reduction rate was 29.9% and its egg reduction rate 13.8% in the test group; 31.9% and 28.0% respectively in the boost group. The egg reduction rate in the boost group was higher than that of the test group (P < 0.05). CONCLUSION: The SjC21.7 nucleic acid vaccine could induce partial protective immunity against Schistosoma japonicum in BALB/c mice. PMID- 12568019 TI - Application of remote sensing for surveillance of snail habitats in Poyang Lake, China. AB - OBJECTIVE: To identify snail habitats and monitor the changes by combining remote sensing technique with the ground-based ecological data. METHODS: Of 100 survey sites selected throughout Poyang Lake, 75 were randomly identified from 574 land mass units: 50 were from snail habitats and 25 were from non-snail habitats based on a historical survey (1982-1984); 25 sites with habitats that did not have snails were also selected. Each site, covering 10,000 m2, was divided into a grid of 100 cells from which 20 cells were randomly selected. Snails, when present, were collected from a 4 m2 frame placed in the center of the selected cell. Satellite Landsat TM images were obtained for the same period as the ground survey data collected. Unsupervised classification was used to classify the images. Identified land-cover types were correlated with snail habitat. RESULTS: The sensitivity and specificity of classified snail habitat were 90.0%-95.6% and 61.1%-68.6%, respectively. Based on the classification, estimated snail habitat areas in Poyang Lake increased from 623.4 km2 in 1999 to 762.8 km2 in 2000. CONCLUSION: Snail habitats are associated with grassland-covered areas. Classified images can be used to identify snail habitats, determine their areas, and monitor snail habitat changes caused by annual fluctuations of key environmental factors. PMID- 12568020 TI - [Effect of pentoxifylline on the expression of hepatic TGF-beta 1, type I and type III collagen in mice with liver fibrosis due to Schistosoma japonicum infection]. AB - OBJECTIVE: To study the effect of pentoxifylline (PTX) on the content of hepatic TGF-beta 1, type I and type III collagen in schistosome-infected mice with liver fibrosis. METHODS: Forty mice with schistosomiasis were divided into four groups: one group as control without any treatment, other three were treated with praziquantel 500 mg/(kg.d) for 2 d, high dose PTX 360 mg/(kg.d) for 8 wk, and low dose PTX 180 mg/(kg.d) for 8 wk respectively. Immunohistochemical technique and multimedia color pathographic analysis system were applied to observe the content of hepatic TGF-beta 1, type I and type III collagen in mice infected with S. japonicum before and after treatment. RESULTS: The effect of PTX on the content of hepatic TGF-beta 1, type I and type III collagen in mice was related to the dosage of PTX. High dose PTX treatment significantly reduced the content of TGF beta 1, type I and type III collagen compared to the control (P < 0.01), whereas no difference was found between the group of low dose PTX treatment and control (P > 0.05). CONCLUSION: High dose PTX treatment could reduce the content of hepatic TGF-beta 1, type I and type III collagen significantly in schistosome infected mice with liver fibrosis. PMID- 12568021 TI - Glutathione inhibits the antischistosomal activity of artemether. AB - OBJECTIVE: To observe the effect of reduced glutathione (GSH) and glutathione depleting agent 1-chloro-2,4-dinitrobenzene (CDNB) on the susceptibility of adult Schistosoma japonicum to artemether (Art) in combination with hemin in vitro. METHODS: In vitro, malondialdehyde(MDA) levels were determined in five-week-old worms incubated without or with Art, hemin, GSH, and CDNB, either alone or in combination, for 24 h, and the remaining worms were continuously incubated up to 96 h for worm survival assessment. In vivo, GSH levels were determined in worms freshly recovered from mice 6, 12 and 24 h after treatment with Art 300 mg/kg. RESULTS: In vitro, GSH decreased the proportion of worms killed by Art plus hemin, but CDNB rendered the worms susceptible to the killing. The above mentioned distinguishing features of GSH and CDNB were associated with their reverse effect on worm lipid peroxidation induced by Art-hemin system. In vivo, Art led to a slight decrease followed by a significant increase in the parasite GSH levels. CONCLUSION: GSH might play an important role in the defense of the worms against Art-generated toxic peroxides and free radicals. PMID- 12568022 TI - [Effect of smear thickness on the morphology of protozoan parasites in specimen preparation]. PMID- 12568023 TI - [Ultrastructural dynamic observation on murine schistosomal hepatic fibrosis]. AB - OBJECTIVE: To explore possible mechanisms of hepatic fibrosis by investigating the ultrastructural dynamic changes of liver tissue, especially several kinds of cells related to hepatic fibrosis. METHODS: Murine schistosomal hepatic fibrosis model was established by infecting mice with Schistosoma japonicum cercariae. Routine transmission electron microscopy was used to observe the liver tissue. H.E. staining was used for examining the pathological changes. RESULTS: H.E. staining showed that the model was established successfully. Ultrastructural observation showed that at the 6th week after infection, the necrosis of hepatocytes around the acute granulomas occurred; the number of sinusoidal endothelial fenestrae and vitamin A droplets in fat-storing cells decreased; large phagosomes and rough endoplasmic reticulum could be seen in the cytoplasm of Kupffer's cells. At the 8th week, steatosis was found in some hepatocytes, some microvilli emerged on a few inter-hepatocytic surfaces and the inter hepatocytic spaces were enlarged. Large collagen fibrillar bundles filled in the perisinusoidal spaces, and capillarization of hepatic sinusoids was observed. Secretory vesicles filled with collagen fibrils appeared in the cytoplasm of fat storing cells with large amount of collagenous fiber bundles surround the cells. Rough endoplasmic reticulum increased in Kupffer's cells. At the 10th week, fat storing cells were activated and transformed into myofibroblasts. At the 12th week, the number of myofibroblasts decreased but that of fibroblasts and fiber cells increased. CONCLUSION: Activation of fat-storing cells and transformation from fat-storing cells into myofibroblasts are the critical link in the development of hepatic fibrogenesis following schistosome infection. Kupffer's cells, necrotic hepatocytes and sinusoidal endothelial cells may relate to the activation of fat-storing cells. Capillarization of hepatic sinusoids possibly accelerates the development of hepatic fibrosis. PMID- 12568024 TI - [Susceptibility and immune responses to Pneumocystis carinii infection in different strains of mice by contagious transmission]. AB - OBJECTIVE: To explore the susceptibility of different strains of mice to P. carinii infection and the host immune response to the organism. METHODS: C57BL/6 and BALB/c strains of mice (15 each) were exposed to SCID mice infected with P. carinii by co-housing. Observations were made on the number of parasites in the lungs, level of CD4+ T cell and CD8+ T cell in BALF, and serum IgG at 4, 5 and 6 wk after contagion. RESULTS: The number of P. carinii grew in the lungs of BALB/c mice was found much greater than those in C57BL/6 mice. A few number of P. carinii cysts were detected in the lungs of both strains of mice by 4 wk after co housing, the number of cysts increased at 5 wk in the lungs of BALB/c mice but not in that of C57BL/6 mice. P. carinii-specific IgG in the sera and high level of CD62low CD4- and CD8-positive T cells in the lungs were found at 5 wk. The parasites were cleared from the lungs at 6 wk in all infected mice, shortly after acquired immune response was initiated. CONCLUSION: BALB/c mice are more susceptible to P. carinii than the C57BL/6 mice by natural transmission, and all the immunocompetent mice can clear the infection of P. carinii by cellular and humoral immune responses. PMID- 12568025 TI - [One case of Pneumocystis carinii infection after renal transplantation]. PMID- 12568026 TI - [Changes of elements in Culex quinque fasciatus larvae after infected by Lagenidium giganteum (Oomycetes: Lagenidiales)]. AB - OBJECTIVE: To analyze the quantitative change of 18 elements in Culex quinque fasciatus Larvae infected by Lagenidium giganteum (Oomycetes: lagenidiales), and to explore possible mechanism of mosquito-killing by the fungus. METHODS: Elements in the mosquito larvae at different developmental stages from infected and non-infected (control) groups were detected with automatic bioassay machine. RESULTS: At the early stage of infection, content of Ca and Cu in the infected group was markedly lower while the content of Co and Cd were higher than the control. In 24 hrs after infection, the content of Ca, Mg, Na, P, Fe, Zn, Cu, Mn and Cr was significantly lower while that of K, Mo, Co, Ni, Al, Cd and Pb were higher than those in non-infected larvae. No marked difference was found for the content of Sr and B between the two groups. In 48 hrs after infection, considerable difference was found in 17 elements between infected and noninfected larvae, with only one exception of Sr. CONCLUSION: Many elements which play an important role in maintaining the vitality of the mosquito larvae decreased and some of the heavy metal elements accumulated after the infection of Lagenidium giganteum. This might be a part of the killing mechanism to Culex quinque fasciatus larvae. PMID- 12568027 TI - [Heterophyes heterophytes in hen's eggs]. PMID- 12568029 TI - [Report on six cases of paragonimiasis]. PMID- 12568028 TI - [Content of nitric oxide in cerebrospinal fluid from patients with neurocysticercosis]. AB - OBJECTIVE: To evaluate the relationship between the concentration of nitric oxide (NO) and cytological change in cerebrospinal fluid (CSF) and the onset of neurocysticercosis in patients. METHODS: NO Level in CSF was detected in 30 cases of neurocysticercosis, 20 cases of tuberculous meningitis (TBM) and 20 healthy people as control by using Griess method. The cytological components in CSF were also examined. RESULTS AND CONCLUSION: Griess method was proved to be a rapid and accurate technique for the detection of NO content in CSF. The NO concentration in cases of neurocysticercosis and TBM was significantly higher than that of control(P < 0.01). The neurocysticercosis cases showed the highest CST NO level which was considerably higher than that of TBM cases(P < 0.05). Among the cytological changes, the neurocysticercosis cases also showed a higher count of eosinophils than that of TBM cases and control (P < 0.01). The NO level and eosinophils count in CSF increased significantly in patients with neurocysticercosis. PMID- 12568031 TI - [Technical measures of filariasis elimination in Tengzhou City]. AB - OBJECTIVE: To summarize the technical measures in achieving the elimination of lymphatic filariasis in Tengzhou city. METHODS: To collect and analyze the materials on research and control of lymphatic filariasis in Tengzhou city during the disease elimination program over the 40 years. RESULTS: Following to the national criteria for the control and elimination of filariasis, efforts were made for epidemiological investigation, mass blood examination, and mass drug administration by taking diethylcarbamazine (DEC) medicated salt for the whole population. After the criteria of transmission interruption (basic elimination) of filariasis were met in 1974, re-examination and treatment for the cases with microfilaremia, and parasitological and entomological surveillance were conducted. In 1995, filariasis elimination in the whole city was declared. CONCLUSION: Comprehensive measures focusing on the elimination of source of infection were proved effective for interrupting the transmission of filariasis. Unremitting surveillance was necessary for achieving the goal of elimination of filariasis. PMID- 12568030 TI - [Observation on moving patterns of Oncomelania snails in rivers and canals]. AB - OBJECTIVE: To understand the moving patterns of Oncomelania snails, intermediate host of S. japonicum, in the water bodies. METHODS: Based on the biological features of the snails, methods and techniques in relation to hydraulics and silt engineering were adopted to investigate the active scrawl ability and passive movement of the snails. RESULTS AND CONCLUSION: The scrawl speed of the snails themselves was very low, 2.45 mm per minute only. The active movement in the water current was therefore almost negligible. The major moving patterns of the snails in rivers and canals were that: 1. Snails adsorbed on different kinds of carriers drifted on the water. 2. Snails suspended on the water and drifted with the current, these were the young snails under 7 weeks of age. Adult snail showed a strong ability of adhesion, 12 times higher than its body weight. PMID- 12568032 TI - [Application of intradermal test for schistosomiasis surveillance in areas where the disease transmissions is interrupted]. PMID- 12568033 TI - [Re-emergence of schistosomiasis in Dali City after the criteria of transmission control were met]. AB - OBJECTIVE: To analyze the epidemic situation on schistosomiasis in five years after reaching the criteria of transmission control in 1995 in Dali City. METHODS: Epidemiological data of schistosomiasis during 1995-2000 were collected and analyzed. RESULTS: Stool positive rate in the population was increasing yearly, new cases of infection including cases under 14 years old were detected each year. Acute infection occurred during 1996-1998 consecutively. Stool positive rate in cattle was unstable. Snail-ridden area increased significantly in 1996 and spots with infected snails stayed at a higher level during 1997-2000. CONCLUSION: Schistosomiasis in Dali city has been re-emerging though it was once deemed under transmission control. Possible reasons include a low drug-taking rate though distributed, frequent movement of cattle population, and less activity on snail control. PMID- 12568034 TI - [Epidemiological trend and characteristics of malaria in Liangping County of Chongqing during 52 years]. PMID- 12568035 TI - [Cost-effectiveness analysis of the current measures for malaria prevention in Yuanjiang valley, Yunnan province]. AB - OBJECTIVE: To evaluate the cost-effectiveness analysis of the measures currently used for malaria prevention in the region of Yuanjiang River Valley. METHODS: The efficacy and cost-effectiveness analysis of chemoprophylaxis, single DDT residual spraying and a combination of both were evaluated through three field intervention trials. RESULTS: (1) From July to November, the rate ratio of malaria incidence among building workers of expressway as the group of chemoprophylaxis using combined piperaquine/sulfadoxine once per month was 0.35 (95% CI: 0.20-0.59); the cost per individual protected (CIP) and the cost per case averted (CCA) were RMB 6.69 yuan and RMB 459 yuan respectively. (2) In the hyper-endemic villages near the river, the rate ratio in the group using chloroquine plus primaquine once every 10 days for 5 months was 0.28 (95% CI: 0.08-0.99); the CIP and CCA were RMB 5.30 yuan and RMB 300 yuan respectively. (3) The standardized morbidity ratio (SMR) was 100%, 98% and 50% respectively for the chemoprophylaxis group with pyrimethamine/sulfadoxine once per month from May to September, the group with single DDT residual spraying in April and the group of combined chemoprophylaxis and DDT spraying. The CIP was RMB 1.49, 2.48 and 3.97 yuan for the three groups respectively. As compared with the previous year, no cases were averted from the chemoprophlaxis group; the CCA was RMB 14,535 yuan and 908 yuan respectively for the DDT spraying group and the group with combined measures. CONCLUSION: There was no significant difference on the chemoprophylaxis cost between the group using piperaquine/sulfadoxine combination once per month and that of chloroquine plus primaquine once per ten days. However, the study proved an efficacy for malaria prevention in the former but not in the latter. In comparison with the groups of single measures, the group with combined measures showed the best efficacy and effectiveness, but highly expensive. PMID- 12568036 TI - [Investigation on acute schistosomiasis in an inland hilly village]. PMID- 12568037 TI - [Applications of bioinformation technology in research of Plasmodium falciparum genome]. PMID- 12568038 TI - [Liposome-mediated delivery system in prophylaxis and therapy of malaria]. PMID- 12568039 TI - [Evaluation on surveillance and intervention of lymphatic filariasis after interruption of the disease transmission in Jiangxi province]. PMID- 12568040 TI - [A fast and simple method for examining helminth eggs]. PMID- 12568041 TI - [Surgical treatment of cerebral cysticercosis with intracranial hypertension]. PMID- 12568042 TI - [CAb and CAg detection in population groups from areas with different endemicity of Schistosoma japonicum]. PMID- 12568043 TI - [Morphological identification of filarial larvae and adults of Strongyloides stercoralis in the sputum of a heavily infected patient]. PMID- 12568044 TI - [An investigation on intestinal parasitic infections among inhabitants in Jiaxing City]. PMID- 12568045 TI - [Investigation of Toxoplasma gondii infected animals in Yunnan province]. PMID- 12568046 TI - [Pulmonary function in 20 patients with hydatis disease]. PMID- 12568047 TI - Modernisation. Alien concept. PMID- 12568048 TI - Modernisation. 'What new NHS?'. AB - A 2001 survey of 203 doctors, nurses and midwives who had spent an average of 15 years in the NHS conducted at a teaching hospital in Belfast, showed poor understanding of the government's 'new NHS' agenda. Seventy-nine per cent were unaware that there was a new NHS. Only 44 per cent felt the principles of the new NHS were of any value in clinical practice. Only 51 per cent could name the health secretary. Much work needs to be done to bring home the importance of the government's quality agenda to clinical staff. PMID- 12568049 TI - Modernisation. Fear of flying. AB - Interviews with senior managers and clinicians involved in the national booking programme and cancer care collaborative found that scepticism, particularly among doctors, was a barrier to implementing change. Regular feedback and clear evidence of change emerged as key factors in persuading sceptics of the benefits of different ways of working. Getting sceptics on board can be a long process and time spent talking to them is not wasted. Consultants' support is key for endorsing change. PMID- 12568050 TI - Addressing London's needs. PMID- 12568051 TI - Advancing to better solutions. PMID- 12568052 TI - Rethinking construction procurement. PMID- 12568053 TI - Putting the patient first. PMID- 12568054 TI - Waste handling excellence. PMID- 12568055 TI - Focus on computer-aided diagnosis. PMID- 12568056 TI - A series of awakenings. PMID- 12568057 TI - Network technology offers significant benefits. PMID- 12568058 TI - Asbestos--why is this subject being dragged up again? PMID- 12568059 TI - Using "short" interrupted time-series analysis to measure the impacts of whole school reforms. With applications to a study of accelerated schools. AB - The present article introduces a new approach for measuring the impacts of whole school reforms. The approach is based on "short" interrupted time-series analysis, which has been used to evaluate programs in many fields. The approach is used to measure impacts on three facets of student performance: (a) average (mean) test scores, which summarize impacts on total performance; (b) the distribution of scores across specific ranges, which helps to identify where in the distribution of student performance impacts were experienced; and (c) the variation (standard deviation) of scores, which indicates how the disparity in student performance was affected. To help researchers use the approach, the article lays out its conceptual rationale, describes its statistical procedures, explains how to interpret its findings, indicates its strengths and limitations, and illustrates how it was used to evaluate a major whole-school reform- Accelerated Schools. PMID- 12568060 TI - Examining the impact of external influences on police use of deadly force over time. AB - Prior deadly force research has sought to identify appropriate mechanisms that can effectively control police officers' decisions to use deadly force. However, the relative impact of external discretion control policies on police shooting behavior remains largely unknown. Using data from Philadelphia for a period of more than two decades, this article employs interrupted time-series analysis (ARIMA) to examine the impact of legislation and judicial intervention on use of deadly force by Philadelphia police officers. The article also considers the impact of larger, community-level characteristics on levels of deadly force. Findings produced modest support for the potential effect of external discretion control policies and community-level factors on police shooting behavior but generally suggest that dynamic changes in the internal working environment can outweigh the influence of the external mechanisms. PMID- 12568061 TI - Methods to reduce the impact of intraclass correlation in group-randomized trials. AB - This study reports intraclass correlation (ICC) for dependent variables used in group-randomized trials (GRTs). The authors also document the effect of two methods suggested to reduce the impact of ICC in GRTs; these two methods are modeling time and regression adjustment for covariates. They coded and analyzed 1,188 ICC estimates from 17 published, in press, and unpublished articles representing 21 studies. Findings confirm that both methods can improve the efficiency of analyses shown to be valid across conditions common in GRTs. Investigators planning GRTs should obtain ICC estimates matched to their planned analysis so that they can size their studies properly. PMID- 12568062 TI - Basic Reading Through Dance program. The impact on first-grade students' basic reading skills. AB - This study evaluates the effectiveness of an arts-based educational program, Basic Reading Through Dance. Basic Reading Through Dance is a 20-session, curriculum-based reading intervention for first-grade students developed by Whirlwind, a not-for-profit organization. There were a total of 721 first-grade students from Chicago public schools who participated in the study, with 328 students from 6 schools receiving the program and 393 students from 9 schools serving as controls. The program was designed to improve reading skills, as assessed by the PhonoGraphix Test, such as code knowledge (alphabet sounds) and phoneme segmentation (separating letter sounds within spoken words). Results suggest that the students who participated in the program improved significantly more than control students on all reading skills that were assessed. PMID- 12568063 TI - [100 years with certificates: Danish Society of Insurance Medicine 1902-2002]. AB - Danish Society of Insurance Medicine was formed in 1902 as an association between the Danish Society of Accident Insurance and the Danish Society of Life Insurance Medicine, both founded a year before. The purpose of the society is to apply and to further the scientific basis of insurance medicine. The meetings also play a role as a forum for discussions. The members of the society are all working part time in an insurance company in Denmark (in contrast to many other countries where it is a full-time job). The members are mainly specialists in medicine and surgery. During the first half life of the society nearly all members were high qualified doctors, i.e. professors and consultants at a university hospital in Copenhagen usually with a doctoral degree. These prominent doctors had a significant influence on insurance medicine in Denmark. The managing directors of the Insurance Companies were invited to the meetings. Many scientific works and textbooks were written on the subject. The society arranges two meetings yearly - one concerning with life insurance and one with accident-insurance medicine. Previously, the question put to the medical adviser was always "Which impairments can still be insured?", but today the question is the opposite "Which impairments are not insurable?" The members have increased to currently 112, still only men, but today the members are younger, comprise fewer professors and fewer with a doctoral degree. During the last 6 years, all interested members in the insurance companies have been invited to the meetings, providing a broader input for the discussions. The topic of the meeting is of current interest: new laws or a new treatment of a disease (medication, surgery) which has been introduced. One to three speakers open the meeting, followed by a discussion. The meetings are always concluded with a light meal and a glass of wine, in earlier days also with cigars and Danish schnaps. 190 meetings have been held during the last 100 years. Often the speakers are members of the society. In the future the Danish Society of Insurance Medicine will face new challenges as regards to the communication of knowledge about new diseases and new treatments in the light of with biostatistics and new insurance products. PMID- 12568064 TI - Microbial ecology comes of age. PMID- 12568065 TI - The cimetidine protocol: a convenient, accurate, and inexpensive way to measure glomerular filtration rate. PMID- 12568066 TI - International activities of the American Society for Microbiology (ASM). PMID- 12568067 TI - Post-streptococcal hypertensive encephalopathy with normal urinalysis. PMID- 12568068 TI - Clinical standards and the structure of professional obligation. PMID- 12568069 TI - Reappraising the role of autonomy in medical ethics. PMID- 12568070 TI - Christian medical and dental associations adopt new bioethics statement: medical malpractice. PMID- 12568071 TI - Dutch physicians: euthanasia has created palliative care deficit. PMID- 12568072 TI - From Slaughter-House to Lochner: the rise and fall of the constitutionalization of public health. PMID- 12568074 TI - Love and death in a pediatric intensive care unit. PMID- 12568073 TI - The use of Zairian children in HIV vaccine experimentation: a cross-cultural study in medical ethics. PMID- 12568075 TI - Whose child am I? A look at how consent affects a husband's obligation to support a child conceived through heterologous artificial insemination. PMID- 12568077 TI - Constitutional mass torts: sovereign immunity and the human radiation experiments. PMID- 12568078 TI - [New screening chest radiography with computer analysis of pulmonary marking movement and change in regional density]. AB - Respiratory function data are useful for the evaluation of lung disease. To develop a new method of computer analysis for screening chest radiography that would provide respiratory kinetics automatically, we quantified and analyzed the movement of pulmonary markings and the changes in lung density during respiration. In this study, we obtained fluoroscopic chest images of a healthy volunteer during respiration using an image intensifier (I. I.) fluoroscopy system. We analyzed the vector of the movement of pulmonary markings and measured the regional mean density of sequential chest images during respiration. The movement of pulmonary markings in the upper lung field was less than that in the lower lung field. Variation in the phase density curve of the upper lung field was less than that of the lower lung field. Our results indicate that changes in density were regular and were synchronized with respiration. This computerized method may aid radiologists in the evaluation of lung diseases such as chronic obstructive pulmonary disease (COPD) , where abnormal respiratory change in density occurs as a result of air trapping. PMID- 12568080 TI - [Noise properties in the fading of the imaging plate on a computed radiography system]. AB - Fading of the imaging plate (IP) in computed radiography (CR) systems with photostimulable phosphors is known well. The relation between noise properties and the fading of the IP in a CR system was investigated. Uniform exposure for the IP with various times between exposure and readout for the same exposure was used for this study. The relative intensity of photostimulated emission and the digital Wiener spectra were calculated. The relative intensity of photostimulated emission decreased with increasing time, whereas Wiener spectral values showed no difference with increasing time. We concluded that noise properties were not affected by the fading of the IP in the CR system. PMID- 12568081 TI - [Application of run length matrix to magnetic resonance imaging diagnosis of Alzheimer-type dementia]. AB - To examine the possibility of diagnosing Alzheimer-type dementia, we studied this condition using the run length matrix, on head MR images of 29 Alzheimer-type dementia patients (8 men, 21 women, 78.7 +/- 6.7 years) and healthy elderly controls (10 men, 19 women, 72.3 +/- 8.7 years) . The results showed that differences in GLN (gray level nonuniformity) and RLN (run length nonuniformity) were statistically significant. Furthermore, discriminant analysis based on GLN and RLN showed a rate of sensitivity of 69.0%, specificity 86.2%, and correct classification 77.6%. Although this rate of correct classification is inferior to the planimetric and volumetric methods, run length matrix is only one method of texture analysis. The results of this study indicate the possibility of MR imaging-based diagnosis of Alzheimer-type dementia with texture analysis including a run length matrix. PMID- 12568082 TI - [Axial calcaneus roentgenograph]. AB - Low-frequency noise caused by intestinal gas, bone, or high-contrast subjects on roentgenographs often makes them difficult to read. The plantodorsal projection of the calcaneus taken by the conventional method has low-frequency noise because of its own high contrast for the long axis. Even if a higher X-ray tube voltage is used to reduce the low-frequency noise, middle- or high-frequency signals such as those of bone trabeculae are also reduced. We investigated a new method that takes into consideration the subject contrast between the part of the posterior calcanean articulation and distal part on the calcaneus projection, the spectrum of bone trabeculae, and the scatter fraction. The technique we introduced has some merit compared with the conventional method in that it reduces image distortion, provides low contrast for the long axis, and sharpens trabeculae images and articulations. PMID- 12568085 TI - "Prenatal adoption": the Vatican's proposal to the in vitro fertilization disposition dilemma. PMID- 12568086 TI - [Examination of MR cholangiopancreatography using the fast recovery single shot fast spin echo sequence]. AB - During this project, we evaluated methods to scan MRCP images with overlapping slice positions during one breath-hold using the FRSSFSE sequence. The FRSSFSE sequence is a technique to arbitrarily change the residual transverse magnetization to longitudinal magnetization. With the SSFSE sequence, the imaging field where the slice position overlaps is subject to substantial influence from the saturation effect of the water component. Therefore, one breath-hold is required for each image. However, the FRSSFSE sequence enabled the deterioration in image quality due to the saturation effect to be minimized even during a short TR. This enabled images of overlapping slice positions to be scanned during one breath-hold. We used a setting of TR=5,000 ms at our hospital, and scanned several images with overlapping slice positions during one breath-hold. The TR=5,000 ms setting was determined from image quality and breath-hold time considerations. The use of the FRSSFSE sequence with MRCP enabled 3-4 images to be scanned at overlapping slice positions during one breath-hold. This method is effective in reducing examination time and the burden on the patient. PMID- 12568087 TI - Family-based informed consent: reply to commentators. PMID- 12568088 TI - When it ain't broke. PMID- 12568090 TI - [Utility of SPECT in gallium scintigraphy]. AB - Whole-body gallium planar scintigraphy is a mainstay for the detection of tumors and inflammatory lesions. Recently, gallium SPECT (single photon emission computed tomography) has become more common in the clinical setting. This diagnostic modality is widely employed in our hospital, and lesions are actually detected by SPECT in some cases. Although the contrast of SPECT images is better than that of planar images, spatial resolution is limited by the limited matrix size. Thus, the overall diagnostic utility of SPECT remains to be confirmed. The usefulness of SPECT for the detection of gallium-accumulated lesions was evaluated in a phantom. In this study, we showed that SPECT is able to detect more smaller and lower gallium accumulations than planar imaging. Thus, SPECT imaging is useful in gallium scintigraphy. PMID- 12568089 TI - Commentary: stalking, threatening, and harassing behavior by patients--the risk management response. PMID- 12568091 TI - [Visualization between popularity and scientific practice: new perspectives for the history of medicine, science, and technology]. AB - The paper investigates how visual resources can be fruitfully used to study the history of science, medicine, and technology from a practical point of view. Two new international and interdisciplinary trends within recent historiography are reviewed: the history of visualisation and the history of popularisation. The results of both trends need to be combined in order to understand the ways in which images of science have been used to communicate science from its place of production within the laboratory to its users within the wider society. From the proposed perspective, visual representations of science (i.e. portraits, images of scientific instruments, measurement results and abstractions) are discussed as a distinct medium in which knowledge producers have transmitted and transformed their findings to the acquirers of knowledge. The paper introduces the wider historiographical framework for a discussion of the following four papers published in this issue of NTM. PMID- 12568092 TI - [Electricity in healing: four different applications in a copper engraving of the Elightenment]. AB - This text describes a single engraving of the picture encyclopedia Bilder Akademie fur die Jugend published from 1780 to 1784. It consisted of 52 picture tableaus, each with nine images that were connected through the biblical topic. The particular image under examination, the "Table 38", shows the healing wonders of Christ, the electrifying machine, a healing physician and the structure of ear and eye. Goal of this text will be to describe the different connections and meanings of these depicted scenes, as in the same time I will argue, that pictures can not only be interpreted by understanding how people looked at them, but also to take into question what people did with them. PMID- 12568093 TI - Evidence based musculo-skeletal treatment. PMID- 12568094 TI - [Romantic medicine in Germany as the philosophical explication for understanding the world and man - Mesmer and mesmerism]. AB - The turn of the 19th century was a rare period in history in which modernisation processes in human knowledge had brought about the rise of new scientific disciplines. They in turn considerably modified the traditional understanding of the world based on a metaphysical-esoteric mode. A multi-dimensional space was opened in research on nature, geography, history and culture as well as in the understanding of man and his spiritual structure. That period marked an important turning point in European thinking also because it expressed a kind of interparadigmatic state, wherein the theories of the old order were no longer applicable to swiftly changing reality. On the other hand, the symptoms of the new order displayed all the shortcomings of still underdeveloped, largely spontaneous attempts at understanding the world on the basis of new theoretical premises. At that time, German medicine, constantly in the process of seeking its theoretical and practical identity, became closely associated with the conceptual context and thinking structure of German idealism. That was an intellectual opposition against the rational and mechanistic model of the world which at the same time affirmed the philosophy of the absolute. In medical theory, it led to the emergence of new pursuits - studies on animal magnetism. Amid all the aberration of German romantic medicine (the ideologisation of and subordination to philosophical principles), its great achievement remains the discovery and attempt to describe areas of man's unconscious existence. PMID- 12568096 TI - Abstracts of the 12th Australian Gynaecological Endoscopy Society annual scientific meeting. 16-17 May 2002, Sydney, Australia. PMID- 12568095 TI - Accommodating medical school faculty with disabilities. AB - More than ten years have passed since the Americans with Disabilities Act (ADA) mandated that all employers provide "reasonable accommodations" for employees with disabilities. This mandate applies to medical schools, but no systematic information is available to assess the accommodations provided to medical school faculty with disabilities. This Issue Brief summarizes anecdotal evidence from several medical schools about the experiences of faculty with disabilities, and the barriers they face in establishing and maintaining their careers. It also recommends practical steps medical schools can take to provide a welcoming and accessible academic medical environment. PMID- 12568097 TI - [Medical myths and notions in Ancient Greece]. AB - The article deals with the views on health and disease prevalent in Ancient Greece, the cradle of modern European medicine, focusing on the ever-present myths functioning in that realm despite attempts to rationally explain medical phenomena. On the basis of the works of Hippocrates and Galen, the author has distinguished five different epistemological attitudes towards those phenomena: the holistic, macrocosmological, monistic, anti-hypothetical and eclectic. The first was based on the idea of mechanical and logical causes. In medicine it is marked by determinism connected with climatic conditions. Hippocrates believed that health depended on the weather, in particular on the effects of winds, types of water and properties of soil. Myth emerged in this conception in the way matter - earth, water, air and fire - was conceived, particular in the properties ascribed to them: cold, humidity, aridity and warmth. The author charges that this conception was permeated with ethnocentrism and cites examples invoked by Hippocrates on the basis of his observations on the Scythians. The macrocosmological attitude involves subordinating medicine to cosmology. Man's body is a microcosm. The author cites the treatise 'On Diets', in which the greatest importance both in the universe and in processes taking place in the human body as ascribed to two factors - fire and water. Their combination was said to have played a crucial role in the typology of corporal and mental constitutions. Those features, together with the seasons of the year, mode of behaviour and food, constitute the four forces guiding vital processes. The author then presents the embryogenic conception contained in the cosmological treatise. It was based on such things as numerological speculations, hence - despite its rationalistic assumptions, consigns it to the mythic. The third attitude, the monistic approach, presents a treatise ascribed to Hippocrates 'On the Sacred Disease' and dealing with epilepsy. The author of the article cites evidence desacralising epilepsy and, by the same token, other diseases. But the treatise stops short of separating medicine from meteorology, as the treatise attempts to present overall phenomena as dependent on one factor - air. The anti hypothetical attitude marks a turning-away from cosmology towards the observation of man as such. Medicine is the art of applying the proper diet according to a given individual's digestive capacity. Nevertheless, this anti-methaphysical medicine creates a fictitious scheme explaining health-related phenomena through the antagonism of two forces: the force of food and the inborn force of the body consumming it. The last attitude- the eclectic approach, is associated with its most distinguished representative, Galen, whose cognitive pursuits combined observation with logic. The author cites Galen's opinions about then current philosophical schools and portrays his method of reasoning and behaviour. But Galen also relied on his imagination with regards to the physiological processes taking place in the human body. That can be illustrated by numerous examples, especially the introduction of the concept of a demiurge, in the author's words - a transcendental craftsman setting the universe in order. The conception made medicine metaphysical once again. In summing up, the author states that Greek authors, despite their attempts at objectivity, became slaves of mythical thinking whenever they tried to explain the invisible. Nevertheless, the significance of imagination, both in the realm of heuristics and in the creation of structures, cannot be denied. Modern medicine also makes use of imagination when faced with the limits of what is available to observation, even though those limits are constantly being extended. PMID- 12568098 TI - Heliox questions. PMID- 12568099 TI - Heliox questions. PMID- 12568100 TI - [A non-classical approach to medical practices: Michel Foucault and Actor-Network Theory]. AB - The text presents an analysis of medical practices stemming from two sources: Michel Foucault's conception and the research of Annemarie Mol and John Law, representatives of a trend known as Actor-Network Theory. Both approaches reveal significant theoretical kinship: they can be successfully consigned to the framework of non-classical sociology of science. I initially refer to the cited conceptions as a version of non-classical sociology of medicine. The identity of non-classical sociology of medicine hinges on the fact that it undermines the possibility of objective definitions of disease, health and body. These are rather approached as variable social and historical phenomena, co-constituted by medical practices. To both Foucault and Mol the main object of interest was not medicine as such, but rather the network of medical practices. Mol and Law sketch a new theoretical perspective for the analysis of medical practices. They attempt to go beyond the dichotomous scheme of thinking about the human body as an object of medical research and the subject of private experience. Research on patients suffering blood-sugar deficiency provide the empirical background for the thesis of Actor-Network Theory representatives. Michel Foucault's conceptions are extremely critical of medical practices. The French researcher describes the processes of 'medicalising' Western society as the emergence of a new type of power. He attempts to sensitise the reader to the ethical dimension of the processes of medicalising society. PMID- 12568101 TI - [Unwanted memory, the Polish eugenic movement in between-the-wars period: side notes to Krzysztof Kawalec's article]. AB - A polemical response to Krzysztof Kawalec's article 'Dispute over Eugenics in 1918-1939', published in 'Medycyna Nowizytna' ['Modern Medicine'], 2000, vol. 7, fascicle 2. In his article Krzysztof Kawalec overlooks the issue of race, which had been at the centre of the eugenic ideology, and then erroneously situates eugenicists on the political spectrum. The eugenicists were not radicals or totalitarians but constituted a group of leftist-liberal intellectuals. Their views were rejected by the Polish government circles in power at that time, not without the deterring influence of Nazi racism and the opposition of the Catholic Church. The main reason why eugenic notions suffered a defeat in pre-war Poland was the isolation and political weakness of eugenic circles. Therefore, issues relating to Polish eugenics during the two decades between the two World Wars should be consigned to a much greater degree to the realm of learning and social movements rather than to the political sphere. PMID- 12568102 TI - Bibliography. PMID- 12568103 TI - [Annual Congress of the American Society of Clinical Oncology, Orlando, Florida, May 2002: update on breast and digestive cancer]. PMID- 12568104 TI - Abstracts of the American Dental Education Association annual session in conjunction with the Association of Canadian Faculties of Dentistry. March 8-12, 2003. San Antonio, Texas, USA. PMID- 12568105 TI - Abstracts of the Annual Meeting of the Society for Adolescent Medicine. March 2003. PMID- 12568106 TI - [The fate of graduates of the Medical Cadet Officers School, the first nine graduating classes (1922-2001): based on the archives of Dr Roman Jakubski]. AB - In 1922 a Military Medical School was set up in Warsaw. Following several reorganisations from 1928 it was known as the Medical Cadet Officers' School. This academy trained physicians, pharmacists and dentists for the needs of the Armed Forces. Professional studies were conducted at the Medical or Pharmacy Faculty of Warsaw University. By 1939, 17 incoming classes had been admitted, amounting to 903 students all told. Diplomas and officer's ranks were awarded to 462 graduates. Another 99 senior students were made officers on the eve of the war. The school ceased to exist the moment World War Two broke out. The subject of this article is the fate of graduates of the Medical Cadet Officers School (admitted from 1922 to 1930) who completed their studies in the first nine graduating classes, spent the war in Poland or returned home after the war. It is based on an analysis of archival materials gathered by the late Dr Roman Jakubski in London, who died in 2000. In 1998, he donated his collections to the Main Medical Library in Warsaw. Many graduates of the Medical Cadet Officers School survived the war, and about 26% of them perished. Many of the latter were murdered by Stalin's NKVD (security police) in Katyn and Kharkov, whilst the remainder died at the hands of the Germans while fighting on all the fronts of World War Two. The overwhelming majority of graduates of the school's first nine graduating classes proved through their often complicated biographies that their Alma Mata had played an important role in their lives. It produced many well trained and at times superbly skilled military physicians. Some of them went on to become outstanding specialists in various fields of medicine and pharmacy. Most of them courageously defended their homeland during the Second War World, providing to be distinguished officers ready to risk their lives when the situation waranted. PMID- 12568107 TI - Medicare physician payment. PMID- 12568108 TI - [State Hygiene Institute, 1918-1954 organisational structure, objectives and tasks]. AB - After Poland regained its independence in 1918, research units were set up to provide scientific assistance to the state health and epidemiological service then being created. Towards the end of 1918 The State Central Hygiene Institute was established and, following successive reorganisations, transformed into the State Hygiene Institute. It operated at Chocimska str. 24 throughout the entire pre-war period, adapting its organisational structure to current needs. As a state public-health institution, it tracked down sources of communicable diseases and the way they were spread, and conducted research to detect cases of Asian cholera and carriers of contagious diseases. It produced vaccinations of other biological products used in the treatment and prevention of epidemics and made a significant contribution to the war on communicable diseases. It also served as the central state institution in charge of inspecting the country's epidemiological condition as well as a scientific-research facility designed to maintain it at a suitable level in accordance with international norms. With various organisational modifications it survived World War Two, its continued existence being maintained by the German occupation authorities. Reactivated after the war it continues to function to this day at its old location - ulica Chocimska 24, where it serves as a scientific-research facility for health and epidemiological services transferred to a separate organisational structure after 1950. Health and sanitary stations became state institutions in charge of anti epidemic campaigns and were responsible for the country's health and epidemiological situation. In 1954, the State Health Inspectorate co-operating with the Ministry of Health was set up. The State Hygiene Institute functions to this day and it is the supervisory, co-ordinating and scientific-research institution of the State Health and Epidemiological Service. PMID- 12568109 TI - [XXVI Congress of the ISIM (International Society of Internal Medicine), Kyoto, Japan, May 26-30, 2002]. PMID- 12568110 TI - Take your endodontist out to lunch: enjoying the referring relationship. PMID- 12568111 TI - The other side. PMID- 12568112 TI - [The patient and medicine in contemporary Kazakhstan, attitudes towards bio medicine and complementary medicine]. AB - The article discusses society's attitudes towards bio-medicine and complementary medicine in Kazakhstan around the end of the 20th century. It presents the transformation of the health-care system in independent Kazakhstan and its influence on the health situation of the population as well as public opinion on bio-medicine. Presented is a broad spectrum of various fields of complementary medicine which achieved great popularity especially during the 1990s. Among the reasons for that growing popularity appears to be public disenchantment with the collapsing state health-care system which is costly and ineffective. At the same time, an important factor is the durability of traditional beliefs of a magical type which is behind the wide use of nonconventional magic-religious practices -- spells and prayers. Overall public socio-economic malaise and a fear of the future are conducive to a general increase of interest in mystical beliefs and occult practices as well as a paranormal phenomena, and the flourishing of complementary medicine naturally fits into that scheme. PMID- 12568113 TI - [European medical history museums: preserving our heritage]. AB - The article focuses on the Polish medical community's lack of interest in its own material medical legacy. In addition, it furnishes examples of museums, collections and exhibitions devoted to the history of medicine in the countries of Western Europe. PMID- 12568114 TI - [Homeopathic collections at the Institute of Medical History of Stuttgart's R. Bosch Foundation]. AB - The author contains the history, review and characteristics of the homeopathic collections kept at the Institute of Medical History in Stuttgart. This is the legacy of Samuel and Melania Hahnemann and other German homeopathic physicians. It encompasses printed texts, documentation of diseases, personal manuscripts and elements of 'homeopathic material culture'. The review of the collections described in the article has been arranged in such a way as to serve as a guide for those researching this area. PMID- 12568115 TI - [A novel viral encephalitis discovered at the end of 20th century--Nipah virus encephalitis]. PMID- 12568116 TI - Transmembrane TNF and IFNgamma induce caspase-independent death of primary mouse pancreatic beta cells. AB - Tumor necrosis factor (TNF) is important in the pathogenesis of autoimmune diabetes. It has an important role in immunological and inflammatory processes, and has also been shown to induce apoptotic cell death. We have shown that TNF + IFNgamma induce islet cell death in vitro. TNF exists as a biologically active transmembrane molecule (tmTNF), which is then cleaved to form soluble TNF (sTNF). We reasoned that sTNF, which has been used in previous studies, may not represent TNF in its physiological form. We compared the contributions of caspase activation and nitric oxide production to beta cell death induced by either tmTNF or sTNF together with IFNgamma. CHO cells transfected with a mutated TNF were used as a source of tmTNF. Either sTNF or tmTNF, together with IFNgamma, induced caspase-dependent cell death of the NIT-1 insulinoma cell line, as measured by DNA fragmentation and a fluorogenic caspase 3 activation assay. TNF + IFNgamma did not induce caspase 3 activation in primary mouse islets. Instead, iNOS gene expression was induced and cell death which was partly NO-dependent occurred. We conclude that the role of TNF in the development of type 1 diabetes is likely to be the activation of gene expression and not apoptosis. It appears that both tmTNF and sTNF act by a similar mechanism to induce beta cell death. PMID- 12568117 TI - TNF-alpha and -beta gene polymorphisms in multiple sclerosis: a highly significant role for determinants in the first intron of the TNF-beta gene. AB - Tumor necrosis factor (TNF)-alpha and TNF-beta are proinflammatory cytokines that have been implicated in the pathogenesis of several autoimmune diseases. The aim of this investigation was to determine whether a determinant in the first intron of the TNF-beta gene (TNF-beta(+252)) and two promoter-region polymorphisms of the TNF-alpha gene (TNF-alpha(-308) and TNF-alpha(-238)) affect susceptibility to multiple sclerosis (MS). DNA samples from 133 Caucasian MS patients and 148 healthy controls from Norway were genotyped for several polymorphic determinants, using polymerase chain reaction-based restriction fragment length polymorphisms (PCR-RFLP) methods. TNF-beta(+252) genotypes were significantly associated with MS: The frequency of TNF-beta 2,2 was increased (p = 0.00009) while the frequency of TNF-beta 1,2 was decreased (p = 0.0012) in MS patients as compared to controls. TNF-alpha genotypes were not associated with MS. These results suggest that the TNF-beta gene plays a significant role in the etiopathogenesis of MS. PMID- 12568118 TI - Decreased IL-4 producing CD4+ T cells in patients with active systemic lupus erythematosus-relation to IL-12R expression. AB - The balance of interferon-gamma (IFN-gamma) and/or interleukin-4 (IL-4) producing T cells and interleukin-12 receptor (IL-12R) expression on T cells were evaluated in patients with active systemic lupus erythematosus (SLE). Assessment of intracellular IFN-gamma and/or IL-4 were conducted with cytoplasmic staining. IL 12R presenting T cells were also assessed by flowcytometry without in vitro stimulation. In SLE, the number of IFN-gamma producing CD4+ T cells was increased, and the absolute number of IL-4 producing CD4+ T cells was significantly decreased. Although the ratio of IL-12R presenting CD4+ T cells was significantly greater, the absolute number did not increase. The ratio of IFN gamma/IL-4-producing CD4+ T cells correlated with the SLE disease activity index (SLEDAI) and was significantly higher among patients with lupus nephritis. Therefore, the imbalance of IFN-gamma/IL-4 producing CD4+ T cells was due to the decrease in IL-4 producing CD4+ T cells and may play an important pathogenic role in active SLE. PMID- 12568119 TI - Stress response of pancreatic islets from diabetes prone BB rats of different age. AB - Protective and/or repair mechanisms are thought to be activated in pancreatic beta cells in response to injury during insulitis. Manifestation of type-1 diabetes may depend on an imbalance between beta cell damage and repair. To prove this hypothesis, the ability of collagenase-isolated islets to respond to heat stress depending on the age of BB rats was investigated. The islets were exposed either to 44 degrees C (HS) or 37 degrees C (control) for 30 min and then kept at 37 degrees C for 5 h. Immediately and 5 h after heat shock, insulin secretion in response to 20 mmol/l glucose and total protein synthesis of heat-exposed islets were significantly diminished as compared with controls. The islet proteins were separated by SDS-PAGE followed by immunoblotting. Islets from BB rats at an age of 6-90 days responded to heat shock with the expression of major heat shock protein 70 (HSP 70). Islets from 3-day old rats, however, did not respond with induction of HSP 70. In contrast we could detect inducible HSP 70 in islets from 3-day old diabetes-resistant LEW rats. In islets from 90-day old BB rats we observed a decreased amount of HSP 70 compared with islets from 9-, 12-, 30- and 60-day old animals. There was also a higher extent of HSP 70 to observe in islets from 90-day old LEW rats as compared with 90-day old BB rats. Differences in HSP 70 expression between islets of 3-day old BB and LEW rats and other age groups of BB rats might represent distinct stages of maturation of islets whereas diminished expression of HSP 70 in islets of 90-day old BB rats at the age of high probability of developing diabetes might result from reduced ability to induce protective mechanisms. PMID- 12568120 TI - Anti-cyclic citrullinated peptide antibodies in patients with juvenile idiopathic arthritis. AB - We analysed the presence of anti-cyclic citrullinated peptide (anti-CCP) and anti keratin (AKA) antibodies of the IgG class in sera of patients with defined juvenile idiopathic arthritis (JIA) of various subgroups with more than one year duration of the disease. Enzyme-linked immunosorbent assay (Immunoscan RA, Eurodiagnostica, The Netherlands) and an indirect immunofluorescence (IIF) test on rat oesophagus substrate (ImmuGloTM, Immco Diagnostics, Buffalo, USA) were used for the detection and quantification of anti-CCP and AKA antibodies in 140 patients with JIA (64 male and 76 female) aged 2-47 years (median 16.5 years). Overall, anti-CCP were found in 7/140 (5.0%) patients including 3/52 RF negative polyarthritis, 2/18 RF positive polyarthritis, 1/15 enthesitis related arthritis and 1/5 unclassifiable arthritis. AKA were detected in 40/140 patients (28.6%, p = 0.04) including 2/11 systemic arthritis, 2/32 oligoarthritis, 18/52 patients with RF negative polyarthritis (34.6%, p = 0.01), 14/18 RF positive polyarthritis (77.8%, p = 0.000002), 2/15 enthesitis related arthritis and 2/3 psoriatic arthritis. While simultaneous negativity for AKA and anti-CCP occurred in most (97/140; 69.3%) studied cases, simultaneous antibody positivity was found only in few (4/140; 2.9%) studied samples. We conclude that while AKA measured using IIF on rat esophagus can be detected approximately in one third of patients with definite JIA with more than 1 year duration of the disease, only rare occurrence of anti-CCP was observed. We conclude that AKA seem to be partly useful to confirm JIA diagnosis, however, useless to follow-up severity or activity in JIA patients. Anti-CCP do not have any additional value in MA cohort in comparison to RA where their diagnostic and prognostic importance was reported. PMID- 12568121 TI - Experimental model for ophthalmopathy in BALB/c and outbred (CD-1) mice genetically immunized with G2s and the thyrotropin receptor. AB - In an attempt to develop an animal model for thyroid-associated ophthalmopathy (TAO) we have genetically immunized BALB/c and outbred (CD-1) mice with cDNAs encoding the thyroid and eye muscle shared protein G2s and full length human thyrotropin receptor (TSHr). Firstly, BALB/c mice were immunized with cDNAs for G2s and the TSHr, alone or in tandem with cDNAs for interleukin (IL)4 or IL12. Control mice were immunized with empty vehicle only. Sera from the great majority of experimental mice contained antibodies against a G2s fusion protein and the flavoprotein (Fp) subunit of mitochondrial succinate dehydrogenase, the "64 kDa protein", with the greatest levels being found at sacrifice (17 wk). Antibody levels in mice immunized with G2s + TSHr or G2s + IL12 were generally higher than those in mice immunized with G2s only. TSHr antibodies (TRAb), measured as TSH binding inhibition, were detected in only two mice. On histological examination of the orbits, mild edema, eye muscle fiber separation and mast cell infiltration in and around the eye muscles were found in the majority of experimental mice, but not in control mice. Splenocytes were transferred from selected G2s-immunized mice to normal syngeneic litter mates. None of the transfer mice had serum antibodies against G2s, Fp or TSHr but their orbital tissue showed the same degree of mast cell infiltration as primary mice. No major histological changes were observed in the thyroid or other skeletal muscle in either primary or transfer mice. Similar results were observed in CD-1 mice although, overall, the model was better expressed than in BALB/c mice. In these mice, serum anti-G2s antibody levels were not significantly different between the various experimental groups except at 16 wk, when they were slightly greater than in control animals. Anti-Fp antibodies were detected at 12, 14 and 16 wk, in all experimental groups, including those immunized with G2s only, and were greatest in mice immunized with TSHr alone. TRAb levels were greatest in mice immunized with both G2s and the TSHr in the presence of TL4, but not IL12. The finding of negative anti-G2s but positive anti-Fp antibodies in some CD-1 mice suggests that eye muscle damage and Fp release must have been mediated by T lymphocytes, rather than antibodies, targeting G2s or some other as yet unidentified cell membrane antigen. Histological changes in the orbit were similar to those observed in BALB/c mice although mast cell numbers were greater, in both primary and transfer mice. Overall, the greatest histological changes were observed in CD-1 mice immunized with both G2s + TSHr + IL4. None of the animals became overtly hyperthyroid or hypothyroid during the course of the study although several of the CD-1 mice had abnormal TSH or T4 levels. These results indicate that we have established a valid model for human ophthalmopathy using the novel thyroid and eye muscle expressed protein G2s, now recognized as a fragment of the winged-helix transcription factor Foxp1, and TSHr, and that G2s and the TSHr are both primary antigens in TAO. Reactivity against a TSHr-like protein may be the first event leading to ophthalmopathy in humans with TAO and experimental mice and eye muscle damage may result from autoimmunity against G2s and Fp as a result of "antigen spreading". PMID- 12568122 TI - Antibody dependent cell mediated cytotoxicity and phagocytosis of senescent erythrocytes by autologous peripheral blood mononuclear cells. AB - Human red blood cells (RBCs) have a life span of 120 days in circulation, after which they are removed primarily by resident macrophages. Autoimmune antibodies are commonly found on effete RBCs and appear to contribute to their removal from the circulation. In this article, we focused on senescent erythrocytes and studied their removal, in comparison to young RBCs, in two RBC-depletion in vitro assays: antibody dependent cell mediated cytotoxicity (ADCC) and erythrophagocytosis. The results were determined prior to and following the addition of anti-D antibodies to the systems. Old (O-RBC) and young (Y-RBC) erythrocytes were separated by differential centrifugation. When incubated with autologous peripheral blood mononuclear cells as attacking cells, both anti-D treated O-RBCs and anti-D treated Y-RBCs were phagocytized and underwent contact lysis. However, O-RBCs had a significantly higher tendency to be phagocytized (p = 0.05) and a higher predisposition to undergo lysis (p = 0.043) than did Y-RBCs. When incubated with attacking cells without anti-D antibodies, O-RBCs were phagocytized while Y-RBCs were not phagocytized at all (p = 0.046). No contact lysis of either source of target cells occurred when incubated with attacking cells alone without anti-D sera. These in vitro results suggest that ADCC may serve as an additional pathway of elimination of senescent erythrocyte in addition to the classical phagocytosis pathway. PMID- 12568123 TI - Salivary glands and noradrenergic transmission in diabetic rats. AB - 1 Type 2 diabetes is associated with diverse oral pathologies in which salivary flow reduction is one of the causes of these oral abnormalities. Scarce literature exists regarding noradrenergic transmission and adrenergic-induced salivary flow in submaxillary and parotid glands of type 2 diabetic rats. 2 We studied noradrenergic transmission as well as the secretory response to alpha1- and beta-adrenoceptor stimulation in the parotid and submaxillary glands of type 2 diabetic rats. 3 Diabetic rats exhibited diminished neuronal uptake, release and endogenous content of noradrenaline (NE) in both salivary glands. Further, NE synthesis was also diminished accompanied by decreased tyrosine hydroxylase activity. Salivary flow responses to alpha1-(methoxamine) and beta-(isoprenaline) adrenoceptor stimulation were reduced in the submaxillary as well as the parotid glands of diabetic rats. 4 Our results suggest that the reduction of noradrenergic transmission in the salivary glands of type 2 diabetic rats is in part responsible for the diminished salivary flow evoked by alpha1- and beta adrenergic stimulation. Reduced noradrenergic activity may contribute to the pathophysiology of oral abnormalities in diabetic patients. PMID- 12568124 TI - Endothelium-dependent relaxation in isolated renal arteries of diabetic rabbits. AB - 1 In this study, we have investigated the vasodilator response to acetylcholine under diabetes conditions in isolated renal arteries of rabbits. We have also examined the contribution of endothelium-derived nitric oxide (EDNO) and endothelium-derived hyperpolarizing factor (EDHF) to the endothelium-dependent relaxation caused by acetylcholine in the renal arteries of alloxan-induced diabetic rabbits. 2 Acetylcholine (10(-10) - 10(-4) M) produced cumulative concentration-response curve in the renal arteries of both control and diabetic rabbits. The EC50 values and maximal responses to acetylcholine were not significantly different relative to diabetic conditions. In order to isolate the EDHF component of acetylcholine-induced vasodilator response, L-nitro-methyl arginine ester (L-NAME, 10(-4) M) and indomethacin (10(-6) M) were added to the Krebs' solution throughout the experiment. Under these conditions, acetylcholine induced vasodilatation in the isolated renal arteries from both control and diabetic rabbits. The vasodilator response to acetylcholine was not affected under diabetic conditions. 3 Sodium nitroprusside (SNP)-induced relaxation was increased in the diabetic rabbits compared with the control animals. 4 Tetrabutyl ammonium (TBA, 0.5 mM) produced a significant reduction in acetylcholine-induced vasodilatation in both preparations from control and diabetic animals, consistent with involvement of K+ channels in mediating this response. Glibenclamide (1 microM) attenuated acetylcholine-induced vasodilatation in preparations from control animals only, while iberiotoxin (0.05 microM) significantly reduced the vasodilator response to acetylcholine in preparations from both control and diabetic animals. 5 The role of EDNO in mediating acetylcholine-induced vasodilatation was examined. The vascular preparations were incubated with 20 mM K(+)-Krebs' solution to inhibit the EDHF contribution to acetylcholine-induced vasodilatation. Under this condition, acetylcholine induced a vasodilator response in both preparations from control and diabetic rats. Pretreatment with L NAME (10(-4) M) attenuated acetylcholine-induced vasodilatation in both preparations, indicating an nitric oxide-mediated vasodilator response. 6 Our results indicated that acetylcholine-induced vasodilatation in the isolated renal arteries of alloxan-induced diabetic rabbits was not affected under diabetic conditions. Acetylcholine-induced vasodilatation is mediated by two vasodilator components; namely, EDHF and EDNO. The contribution of EDHF and EDNO to acetylcholine-induced vasodilatation was not affected under diabetic conditions and there was no indication of endothelial dysfunction associated with diabetes. EDHF component was found to act mainly through high conductance Ca(2+)-activated K+ channels under normal and diabetic conditions, while the adenosine triphosphate-dependent K+ channels were involved in mediating acetylcholine vasodilator response in the control preparations only. PMID- 12568125 TI - The enhanced plasma levels of soluble tumor necrosis factor receptors (sTNF-R1; sTNF-R2) and interleukin-10 (IL-10) in patients suffering from chronic heart failure are reversed in patients treated with beta-adrenoceptor antagonist. AB - 1 Cytokines may parallel or regulate the beneficial effects of beta-adrenoceptor antagonist treatment observed in chronic heart failure (CHF) patients. Therefore, this study was performed in order to investigate alterations of cytokine levels in beta-blocker-treated patients suffering from CHF. 2 We investigated plasma cytokine levels in eight healthy controls and 12 CHF patients. The patients were treated with standard medication (CHFstd) or with standard medication and additional beta1-blocker metoprolol (CHFmet). Interleukin-(IL)-1alpha, IL-1beta, IL-1 receptor antagonist (IL-1ra), IL-2, IL-6, IL-8, IL-10, tumor necrosis factor alpha (TNF), soluble TNF receptor type 1 (sTNF-R1), sTNF-R2, and sCD14 were measured by ELISA. 3 IL-1alpha and IL-1beta were not detectable in any of the tested groups. IL-2, TNF, or sCD14 were not altered as compared with healthy control subjects. CHFstd patients expressed enhanced IL-1ra, IL-6, IL-8, IL-10, sTNF-R1 and sTNF-R2. In CHFmet patients IL-1ra, IL-6 and IL-8 remained at the same level. In contrast, sTNF-R1 levels were significantly reduced, although not to control, whereas the sTNF-R2 and IL-10 were reduced to control levels. 4 The cAMP levels of mononuclear cells--recalculated for the patients included in this study from previous work [Werner et al. (2001). Basic Res. Cardiol., 96, 290]- correlated inversely with the sTNF-R2 data (Pearson, r = -0.46; P = 0.041; Spearman, r = -0.64, P = 0.002). 5 The present data indicate an interaction of the neurohumoral and the cytokine system in CHF patients at the cAMP level. Thus, measurement and correlation of sTNF-R2 and cAMP may provide a tool useful during investigation of beta-blocker therapy. PMID- 12568126 TI - Inhibitory effects of cortical steroids and adrenocorticotropic hormone on catecholamine secretion in guinea-pig perfused adrenal glands. AB - 1 We investigated the effects of exogenously applied steroids and endogenously released cortisol on catecholamine (CA) secretion induced by cholinergic agonists in perfused guinea-pig adrenal glands. 2 Acetylcholine (ACh) and electrical stimulation induced CA secretion, which was reversibly inhibited by cortisol. Adrenocorticotropic hormone (ACTH) increased the concentration of cortisol in the perfusion effluent and partly inhibited the secretory response to ACh. 3 Cortisol or aldosterone dose-dependently inhibited secretory responses to nicotine and muscarine. These inhibitory effects were not antagonized by mifepristone and spironolactone, respective cortisol and aldosterone receptor blockers. 4 Dexamethasone, cortisone, corticosterone, 11-deoxycortisol, 11 deoxycorticosterone, prednisolone and cholesterol inhibited nicotine-evoked CA secretion. The secretory response to muscarine was inhibited by these compounds except for dexamethasone and prednisolone. 5 Dexamethasone, cortisol and aldosterone had no effect on CA secretion induced by high KCl. 6 These results suggest that steroids affect nicotinic and muscarinic ACh receptor-mediated responses through distinct mechanisms, and that cortisol released from the adrenal cortex inhibits CA secretion from the adrenal medulla. PMID- 12568127 TI - The effect of nitric oxide synthase blockade on responses to morphine in rat aortic rings. AB - 1 It has been suggested that opioids may play an indirect role in the regulation of the peripheral circulation through the control of nitric oxide (NO) release in vascular tissue. The current study was undertaken to investigate the effect of nitric oxide synthase (NOS) blockade on responses to morphine in phenylephrine (PE)- or KCl-precontracted rat aortic rings. 2 Morphine (3 x 10(-8) - 3 x 10(-5) M) administration did not cause any significant effect on basal tonus of endothelium-intact or endothelium-denuded preparations. Morphine produced concentration-dependent relaxation responses in endothelium-intact as well as in endothelium-denuded rat aortic rings precontracted by PE or KCl. Removal of endothelium did not significantly alter the relaxation responses to morphine. 3 The relaxant responses to morphine were significantly and partially inhibited by pretreatment of tissues with naloxone (NAL, 3 x 10(-5) M) for 5 min. The inhibitory effect of NAL on relaxant responses to morphine in PE- or KCl precontracted rings did not differ significantly between endothelium-intact and endothelium-denuded preparations. 4 Incubation of endothelium-intact or endothelium-denuded rat aortic rings with NOS inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME, 10(-4) M) for 20 min did not cause a significant inhibition on relaxation responses to morphine. 5 These findings confirmed the presence of opiate receptors in rat thoracic aorta, but suggested that mechanisms other than NO release play a role in the relaxant effect of morphine on rat aortic rings. PMID- 12568128 TI - Induction of duodenal motility activates the sphincter of Oddi (SO)-duodenal reflex in the Australian possum in vitro. AB - 1 The aim of this study was to determine if stimulation of duodenal motility by duodenal fluid distension or by administration of carbachol, activates the sphincter of Oddi-duodenal reflex, in an in vitro preparation from the Australian possum. 2 Duodenal distension was achieved by infusion of Krebs solution (0-8 cm H2O). In separate experiments, the sphincter of Oddi (SO) was partitioned from the duodenum and carbachol (10(-7) - 5 x 10(-6) M) added to the duodenal compartment. 3 Fluid distension increased duodenal motility to 120-600% of control activity. These treatments induced increased SO motility (to 120-390% of control) in six preparations, reduced activity (to 60% of control) in one and no response in another. 4 Addition of carbachol to the duodenal compartment resulted in increased duodenal motility. SO motility was increased in seven preparations, reduced in another two and no response were evoked in two others. All SO responses were blocked by tetrodotoxin pretreatment. 5 These data suggest that the SO receives inputs from duodenal mechano and/or stretch receptors resulting in excitatory or inhibitory responses, with the excitatory response dominating. These findings support the role for the SO-duodenal reflex in preventing duodenobiliary/pancreatic reflux during periods of elevated duodenal activity. PMID- 12568129 TI - Neuropeptide Y inhibits double peaked vasoconstrictor responses to periarterial nerve stimulation primarily through prejunctional Y2 receptor subtype in canine splenic arteries. AB - 1 The effects of BIIE 0246, a novel and non-peptide neuropeptide Y (NPY) Y2 receptor antagonist on sympathetic vasoconstriction of the canine splenic artery were investigated. 2 The vasoconstrictor response to periarterial electrical nerve stimulation was described to be a double peaked vasoconstriction consisting of an initial transient, dominantly P2X purinoceptor-mediated constriction followed by a prolonged, mainly alpha1 adrenoceptor-induced response. 3 BIIE 0246 at a concentration of 0.1-1 microM dose-dependently potentiated double peaked constrictions at low frequencies (1 and 4 Hz), whereas at high frequency (10 Hz), it failed to affect these responses. BIIE 0246 (1 microM) also enhanced double peaked responses even in the presence of rauwolscine (0.1 microM). NPY (13-36) (1 100 nM), a selective Y2 receptor agonist reduced these two peaked responses in a dose-related manner. The vasoconstriction to noradrenaline (0.1-10 nmol) or adenosine triphosphate (0.01-1 micromol) was not significantly influenced by either 1 microM BIIE 0246 or 100 nM NPY (13-36). Exposure of tissues to 1 microM BIIE 0246 almost completely prevented the suppression of double peaked constrictions by NPY (13-36) (10 nM) or by NPY (10 nM). 4 We conclude that NPY inhibits sympathetic purinergic and adrenergic vasoconstrictions through an activation of prejunctional Y2 receptor subtype in the neurovascular junction of the canine splenic artery. PMID- 12568130 TI - The effects of hypochlorite-induced oxidative stress on presynaptic M2-receptors at sympathetic nerve endings in the rat tail artery. AB - 1 It was shown recently that stimulation of cardiac muscarinic M2-receptors revealed an enhanced negative inotropic response in isolated rat left atria after exposure to hypochlorite-induced oxidative stress. This phenomenon was not observed after stimulation of the cardiac A1-receptor, which like the M2-receptor is coupled to Gi-proteins. Since even the contractile response to M3-receptor stimulation was not amplified in the rat portal vein, we hypothesized a M2 receptor specificity of this hypochlorite-induced enhancement. 2 The present study was performed in order to investigate whether the sympathoinhibitory response to presynaptically located M2-receptor stimulation would also be modified after exposure to hypochlorite in the rat tail artery. We applied electrical field stimulation (EFS) in order to mimic sympathetic neurotransmission. 3 EFS increased the vascular tone frequency-dependently (0.3-4 Hz). EFS-induced vasoconstriction could be attenuated by acetylcholine (30 nM-1 microM) in a concentration-dependent manner. Hypochlorite (10 and 100 microM) did not affect the sympathoinhibitory effect of acetylcholine (100 nM). 4 In conclusion, in contrast to cardiac M2-receptors, hypochlorite did not amplify the sympathoinhibitory effects of presynaptic M2-receptors. The different responsiveness between neuronal and cardiac M2-receptors to hypochlorite may be explained by the different G-protein subunits involved in the activation of the underlying signalling cascade. PMID- 12568131 TI - Increase in twitch force of the adductor pollicis muscle with stabilized preload at constant thumb abduction before and after administration of muscle relaxant. AB - OBJECTIVE: To determine whether the twitch force of the adductor pollicis remains stable when 0.1 Hz single twitch stimulation is started after stabilization of the thumb preload at a constant degree of thumb abduction; also to study any possible increase in twitch force before the onset of and after the recovery from neuromuscular block. METHODS: Measurements were performed in thirty patients under general anaesthesia. Twitch forces were first allowed to stabilize after allowing the preload to drift to its resting tension at a constant degree of thumb abduction. Three groups of ten patients then each received either vecuronium (2, 4, 8, 16 and 32 microg/kg(-1), successively at intervals of 2 min), d-tubocurarine (5, 10, 20, 40 and 80 microg/kg(-1), successively at intervals of 2 min), or suxamethonium (0.025, 0.05, 0.1, 0.2 and 0.4 mg/kg(-1), successively at intervals of 2 min). Measurements were continued until twitch forces had recovered from neuromuscular block and were stable. RESULTS: Twitch forces stabilized at 114% (sd = 8.9) of the initial value after 10.9 (6.1) min of stimulation. Increase in twitch force before the onset of neuromuscular block was seen in two patients receiving vecuronium and in two patients receiving d tubocurarine. Increase in twitch force after recovery from neuromuscular block was seen in all patients receiving suxamethonium. CONCLUSIONS: Twitch forces may increase when stimulation is started after stabilization of thumb preload at a constant degree of thumb abduction. In some patients twitch forces may increase before the onset of neuromuscular block with vecuronium or d-tubocurarine; twitch forces increase after recovery from suxamethonium. PMID- 12568132 TI - Nonlinear model-based predictive control of non-depolarizing muscle relaxants using neural networks. AB - Neuromuscular blockade can be relatively easily measured in the clinical setting. Consequently, closed-loop control can be exercised by measuring the neuromuscular activity, calculating the dose of drug necessary to achieve a predefined degree of neuromuscular blockade and finally directing an infusion pump. Recently introduced short-acting blocking agents like mivacurium provide benefits for the clinical routine due to a small onset time and half life. In order to provide a stable blockade for different groups of patients a fast and highly adaptable control unit is needed. Furthermore its development should not imply costly investigations for determining a pharmacological model. The fulfilling of these requirements yield a self-adapting model-based predictive control system. The application of artificial neural networks allows an appropriate adjustment of specific parameters without the knowledge of inner pharmacodynamic processes. In a clinical study the EMG module within a Datex AS/3 monitor was used to measure the blockade and a Grasepy 3500 infusion pump for i.v. administration of mivacurium to 35 patients (ASA I-III). The performance of the novel system (mean of the T1 error: -0.32 +/- 1.7) compares favourably with closed-loop controllers demonstrated in the past. These promising results and the easy adaption to other blocking agents encourage to apply this technology even for delivering hypnotic drugs. PMID- 12568133 TI - Lithium dilution cardiac output measurements using a peripheral injection site comparison with central injection technique and thermodilution. AB - OBJECTIVE: The lithium dilution technique for the measurement of cardiac output by the central injection of lithium chloride was introduced by Linton et al. in 1993. In the present report, we compare lithium dilution cardiac output measurement (LD) by the peripheral injection of lithium chloride (pLD) and by central venous injection (cLD), cardiac output determined by electromagnetic flowmetry (EM), and conventional thermodilution cardiac output measurement (TD) on ten swine. METHODS: The animals were monitored with a pulmonary artery catheter, a femoral artery catheter, and an electromagnetic flowmeter placed around the ascending aorta. cLD, pLD, TD, and EM were determined at the baseline, then in a hyperdynamic state produced by dobutamine administration, at a second baseline, and finally in a hypodynamic state induced by propranolol during deep anesthesia. Data were analyzed by linear regression analysis and the comparison method described by Bland and Altman; bias and precision were calculated using the method of Sheiner and Beal. RESULTS: The correlation coefficient between pLD and EM (0.86) was significantly less than that between cLD and EM (0.96), however it was not significantly different from that between TD and EM (0.85). The precision value of pLD (0.14) was the same as that of TD (0.14). CONCLUSION: The results of the present study indicate that pLD is a reliable technique. PMID- 12568134 TI - Waste gas monitor reduces wasted volatile anesthetic. AB - OBJECTIVE: The increasing focus on health care costs requires that all physicians evaluate practice behaviors. The primary emphasis in anesthesia has been limiting the use of expensive medications and interventions. Reducing waste is another approach, and volatile anesthetics are an appropriate target in that simple reduction of fresh gas-flow (FGF) rates is effective. A monitor that measures and displays the cost of wasted volatile anesthetic was developed and used to determine if real-time display of the cost would result in decreased FGF rates, which, in turn, would decrease wasted anesthetic. METHODS: The waste gas monitor (WGM) measures flow rate at the anesthesia machine's scavenger port, integrates this with agent concentration, and displays the calculated cost, real-time, on a portable computer screen. The WGM equipment was attached to the anesthesia machine in the gynecologic surgery operating room (OR) and those cases performed under general endotracheal anesthesia and lasting longer than one hour were eligible for inclusion. First year anesthesiology residents assigned to the study OR as part of a non-specialty rotation, were the subjects of the study. For each resident, after data were collected from at least two eligible baseline cases (Baseline Phase, WGM not visible and resident unaware of its presence), the monitor was introduced and data collection continued for at least three more eligible cases (Visible Phase). RESULTS: Nine residents were initially enrolled, but due to scheduling difficulties only five residents completed the protocol. Data from cases using the WGM demonstrated a 50% decrease (3.58 +/- 1.34 l/min vs. 1.78 +/- 0.51 l/min (p = 0.009)) in the scavenger flow rates, which resulted in a 48% ($5.28 +/- 0.68 vs. $2.72 +/- 0.80 (p = 0.002)) decrease in hourly cost of wasted volatile anesthetic. There was no difference between the Baseline and Visible phases with regard to use of nitrous oxide or intravenous anesthetic agents. CONCLUSIONS. The WGM decreased wasted volatile anesthetic by encouraging decreased FGF rates. PMID- 12568135 TI - Evaluation of a new fibre-optical monitor for respiratory rate monitoring. AB - OBJECTIVES: To present further development of a fibre-optical respiratory rate monitor and evaluate the function in the final version. METHODS: Respiration was recorded in 18 healthy volunteers of ages 9-83 years by three different methods simultaneously: fibre-optical monitoring, capnography and manually. The recordings were made-breath by breath in four 3-minute periods in each subject. In total there were 54 observations, corresponding to 648 minutes. The recordings were fed simultaneously into a computer. Two subjects were excluded because of pronounced differences between all three methods. One person was later diagnosed as having a nasal septum deviation. Recordings with any kind of technical problem were also excluded. RESULTS: In total 516 minutes were analysed. Students' paired t-test showed a mean of 0.5 and 0.3 more breaths were recorded by fibre-optical monitor compared with the manual method and capnography respectively. The 95% confidence interval of the differences between the fibre-optical monitor and the other two methods was -0.5 to +1.5 when the mean respiratory frequency was 14 breaths per minute. Analysis of variance revealed that the fibre-optical monitor tended to overestimate the mean respiratory rate (p < 0.001). CONCLUSION: The fibre-optical monitor has sufficient accuracy for clinical monitoring. It is easy to use and inexpensive. Combinations with pulse oximetry seem attractive, to use especially for application outside "high tech areas," for example when respiratory depressant drugs are used or in ambulances. PMID- 12568136 TI - Predicted sevoflurane partial pressure in the brain with an uptake and distribution model comparison with the measured value in internal jugular vein blood. AB - OBJECTIVE: In order to predict the partial pressure of volatile anesthetics in brain tissue, we developed a pharmacokinetic simulation model suitable for real time application. The accuracy of this model was examined by comparing the predicted values against measured values for blood sampled from the internal jugular vein, which was used as a measure of the partial pressure in the brain. METHODS: Our model consists of six compartments: alveoli, arterial blood, a group of vessel-rich organs (VRG), muscle, fat, and venous blood. A volatile anesthetic, sevoflurane partial pressure in each compartment were calculated using the parameters of volume, blood flow, and solubility for each tissue as reported in previous studies. Simulated sevoflurane partial pressures in VRG were considered to reflect those in the brain. We studied 11 patients undergoing elective abdominal surgery or mastectomy. Sevoflurane was maintained at a concentration of 3% (by vaporizer setting) for 25 min. Sampling points were at 0 min (before sevoflurane administration) and 1, 2, 4, 9, 16, and 25 min after the start of inhalation. We measured the sevoflurane partial pressure in inspiratory gas (PI(S)), in end-expiratory gas (PET(S)), in arterial blood (Pa(S)), and in internal jugular vein blood (Pj(S)). These values were compared against those for the simulated brain (PB(S)sim). RESULTS: The sevoflurane partial pressures increased, in order from least rapid to most rapid, as follows: Pj(S), PB(S)sim, Pa(S), PET(S), and PI(S). The differences between Pj(S) and PB(S)sim were significantly smaller than those between Pj(S) and PET(S) at all sampling points. PB(S)sim did not differ significantly from Pj(S) at any sampling points after 4 min of inhalation, while PET(S) differed significantly from Pj(S) at all sampling points. CONCLUSION: We conclude that our model is clinically useful for predicting sevoflurane partial pressure in the brain, assuming that Pj(S) reflects the sevoflurane partial pressure in the brain. PMID- 12568137 TI - High fidelity correction of pressure signals from fluid-filled systems by harmonic analysis. AB - Fluid-filled systems are generally used for invasive pressure measurements in cardiology, anesthesiology and intensive care medicine. Wave reflection and attenuation cause considerable signal distortion. METHODS: The transducer signal is amplified (no filtering) and sampled (rate 1 kHz) using an autocorrelation based algorithm to detect instantaneous cycle length. A digital Fourier transformation (DFT) for each heart cycle is performed. Amplitude and phase distortion are corrected using data matrices determined in in vitro experiments or calibration measurements for each fluid-filled system to be used. As a measure for accuracy the maximum of the difference of reference and corrected pressures (DIFF) was selected. 960 analyses were performed to assess the impact of correction, used system, mean pressure, time and A/D sampling rate on the agreement with reference pressure. Clinical examples are presented. RESULTS: Mean pressure was correlated with DIFF (r = 0.83). The correction algorithm achieves a significant (p < 0.001) reduction of DIFF from 20-30 mm Hg to 0-5 mm Hg in the high pressure range and from 1-3 mm Hg to 0-1.5 mm Hg in the low pressure system in in vitro experiments and in clinical pressure recordings. Sampling frequency < 1 kHz reduces accuracy. CONCLUSIONS: High fidelity correction of pressure signals from fluid-filled systems by harmonic analysis is feasible. PMID- 12568138 TI - Anesthesia equipment and human error. PMID- 12568140 TI - The new NHS--friend or foe of complementary therapies? PMID- 12568139 TI - Inhaled nitric oxide delivery and monitoring. AB - Inhaled nitric oxide (NO) was found to cause selective pulmonary vasodilation in the late 1980's and since then there has been a huge interest in studying its clinical benefits. The equipment used to deliver and monitor inhaled NO has gone through a dramatic evolution from simple flow meters and industrial monitors to to-days purpose built, fully integrated, NO delivery and monitoring systems that were designed specifically for the demanding area of the intensive care unit. This paper explores the evolution of inhaled NO delivery systems and identifies the design challenges, the safety and regulatory requirements and the ease of use issues that had to be solved to bring this new exciting new class of medical device in to clinical use. PMID- 12568141 TI - Complementary therapy involvement of physicians: implications for practice and learning. AB - OBJECTIVE: We studied physicians' current and desired clinical role functions within the complementary health paradigm and their perceptions of the necessary educational programs to support them. DESIGN AND SETTING: A questionnaire to determine clinical activities within different complementary therapies was developed and mailed-out to 837 eligible physicians in Hamilton, Ontario, Canada, using a modified Dillman approach. RESULTS: The overall response rate to the mailed survey was 49.8% (417/837), with response rates of 50.2% (115/229) for family physicians and 49.7% (302/608) for specialists. The amount of interactions around complementary therapies between physicians, their patients and complementary therapists appears to be low. At the same time, there is a growing interest among physicians about complementary therapies, particularly with respect to developing their knowledge about efficacy and enhancing their skills in assessment and counselling. CONCLUSIONS: The differential levels of acceptance of different therapies by physicians will influence integration of complementary therapies in mainstream medicine. PMID- 12568142 TI - Cross-cultural differences in GPs' attitudes towards complementary and alternative medicine: a survey comparing regions of the UK and Germany. AB - OBJECTIVE: To investigate whether there is a difference in general practitioners' attitudes towards CAM in the UK and Germany. STUDY DESIGN: A descriptive questionnaire was developed and sent to 97 GPs in the UK and 99 GPs in Germany. RESULTS: The overall response rate was 68%. German GPs showed a (non-significant) overall more positive attitude towards CAM than did British GPs. British GPs made more referrals to complementary practitioners. The most popular CAM therapies that UK GPs referred their patients to were chiropractic treatment, acupuncture and osteopathy. German GPs referred their patients mainly to acupuncture treatment, chiropractic treatment and herbal medicine. A significantly higher number of German GPs reported having practised as a CAM practitioner before and having personally used CAM themselves. Seventy percent of British GPs and 76% of German GPs thought it is safe to prescribe complementary medicine and therapies to patients. CONCLUSION: There are small national differences in referring patients to various CAM modalities. Both nations have an overall positive attitude toward and a high interest in CAM. Lack of scientific evidence and information on training opportunities were important points that were continuously raised by GPs in both countries. PMID- 12568143 TI - Changes in public awareness of, attitudes to, and use of complementary therapy in North East Scotland: surveys in 1993 and 1999. AB - OBJECTIVE: To assess changes in awareness of, use of, attitudes to, and opinions about complementary and alternative medicine (CAM) amongst residents of North East Scotland and to obtain details about CAM use from respondents. STUDY DESIGN: Population survey carried out in 1999, 6 years after the initial study. Postal survey to 800 people to examine eight CAMs; acupuncture, aromatherapy, chiropractic, herbalism, homeopathy, hypnotherapy, osteopathy, and reflexology. RESULTS: A total of 432/800 (54%) responded, of whom 175 (41%) had used at least one type of CAM compared to 29% in 1993. Increases in use were statistically significant for aromatherapy (18% versus 9%), acupuncture (10% versus 6%) and reflexology (9% versus 3%). A greater proportion of 1999 respondents thought CAM should be available on the NHS but a smaller proportion of respondents had concerns about using CAM (25% in 1993 and 20% in 1999). Overall concerns about effectiveness of therapies had increased from 36 to 45%, but fewer individuals were concerned about the cost of therapy in the 1999 survey (52% in 1993 to 22% in 1999). A total of 175 individuals provided details about one CAM they had used. The self-reported primary reasons for using CAM were relief of pain due to headaches or musculoskeletal problems, and for relaxation and relief of stress. The majority of CAM was therapist administered (103/166) as opposed to a bought product. Effectiveness ratings were self-reported but overall 80/166 found CAM very effective and 62/166 partially effective. A total of 65% had consulted their GP about their health problem before using CAM, 59/157 indicated their GP knew they were using CAM and of these, 14 indicated their GP was administering the therapy. CONCLUSIONS: The study has provided further baseline data on which to assess trends in CAM use and highlighted issues for patients and the NHS about the use of CAM to relieve health problems. Results indicate a greater proportion of the population of North East Scotland are both aware of and using CAM to relieve health problems. More research into the implications for the NHS of concurrent use of CAM with conventional medicine is required. PMID- 12568144 TI - A survey of self-medication practices and perceived effectiveness of glucosamine products among older adults. AB - OBJECTIVES: The purpose of this study was to investigate the decision-making and self-treatment practices of older users of glucosamine products, namely (1) how do older adults decide to use glucosamine products? (2) what influences their choice? (3) how do they manage their glucosamine treatment? and (4) what is their perception of the effectiveness, side effects, and costs of glucosamine products? DESIGN: A descriptive, exploratory study using a questionnaire survey. SETTING: Province-wide. Invitations to participate were extended to all people living in a Canadian province who used/had used glucosamine products. RESULTS: Advice from friends was a major factor in the decision to take glucosamine. Cost was a major factor influencing the type and brand name of the product used. Ninety-two percent managed their own treatment. The majority (88%) used glucosamine regularly; only one-third took the recommended daily dose. Sixty-seven percent perceived the product to be helpful in relieving symptoms. CONCLUSION: Public education and further research is needed regarding the use of glucosamine products. PMID- 12568145 TI - Effects of electro-acupuncture on psychological distress in postmenopausal women. AB - OBJECTIVES: To evaluate effects of electro-acupuncture (EA) on general psychological distress and relate to experience of climacteric symptoms in 30 postmenopausal women. DESIGN: A randomised single-blind controlled design was used to evaluate effects of EA and extremely superficial needle insertion, with the latter serving as a near-placebo control. SETTINGS: The Linkoping University Hospital in Sweden. INTERVENTIONS: Fourteen treatments during 12 weeks with follow-ups at 3 and 6 months. OUTCOME MEASURES: General psychological well-being, mood and experience of climacteric symptoms. RESULTS: Mood Scale improved only in EA group and not until 12 weeks compared to baseline, from 110 to 129 (P = 0.01), and to 120 at 3-month follow-up (P = 0.04). Mood was significantly better than control at 8 (P = 0.05) and 12 weeks (P = 0.01). Visual analogue scale estimation of climacteric symptoms was decreased at 4 weeks in both groups, and lasted throughout the study period, in EA group from 5 to 2 (P = 0.04) and in control group from 5 to 3 (P = 0.02) at 6-month follow-up. Well-being was ameliorated from 4 weeks in EA and from 8 weeks in control group until end of study (P = 0.01, P = 0.03). No significant differences on climacteric symptoms or well-being existed between the groups. CONCLUSIONS: This study does not show that EA is better than superficial needle insertion for the amelioration of general psychological distress and experience of climacteric symptoms in women with vasomotor symptoms after menopause. However, the more pronounced effect on mood suggests that EA might have additional effects compared with superficial needle insertion. PMID- 12568146 TI - The effects of a low frequency acoustic waveform on peripheral vascular disease: a pilot study. AB - OBJECTIVE: To evaluate the effects of a low frequency acoustic waveform on peripheral vascular disease (PVD). DESIGN: Pilot study utilizing a one-group pre intervention, post-intervention design. SETTING: Adults with peripheral vascular disease were recruited through local advertisements. The study was conducted at a local facility housing the electroacoustic transducer. INTERVENTION: A 25-min exposure to an electroacoustic transducer. OUTCOME MEASURES: Pre- and post measurement of Doppler ultrasound blood-flow velocities in 10 arteries, ankle brachial index (RBI), foot assessment, and 1-week post telephone survey. RESULTS: A significant increase was noted in the right ankle brachial index (RABI) but not the left. Blood flow increased in all arteries, significantly in four. Thirteen participants reported improvement in symptom of peripheral vascular disease over the following week. CONCLUSIONS: While conclusions must be viewed cautiously, the significant differences noted warrant further study to examine effects of acoustic waveforms on peripheral vascular disease. PMID- 12568147 TI - Food or medicine? Decision time in Europe. PMID- 12568148 TI - CAM and the desperate call for cancer cures and alleviation what can websites offer cancer patients? PMID- 12568149 TI - Complementary medicine in primary care: time to decide. Royal Society of Medicine, London 27th March 2002. PMID- 12568150 TI - The art and science of making choices. PMID- 12568151 TI - Impact factors: arbiter of excellence? PMID- 12568152 TI - Librarian supply and demand. PMID- 12568153 TI - "Mapping the literature" useful for collection development. PMID- 12568154 TI - A job with a view: perspectives from the corporate side of the hospital. AB - A change in job responsibilities from library manager to hospital administrator provides this year's Doe lecturer the opportunity to reflect on the values of the library profession from a fresh perspective. Librarians play a unique role and remain vital to the health care enterprise but are frequently misunderstood. Their role can be viewed from three angles: service, technology, and a unique sort of professionalism. Librarians must focus their service priorities on the needs of the institution, while remaining true to their own unique professional values. They must be advocates for the appropriate use of technology in support of those service roles. The passion that many librarians bring to their jobs makes librarianship a vocation as much as a profession. The mission and vision developed by the American Society of Health-System Pharmacists in 2001 provides a useful model for defining a personal professional mission and vision. PMID- 12568155 TI - Brandon/Hill selected list of print books and journals in allied health. AB - This list of 434 books and 79 journals is intended as a selection guide for print literature to be used in a library supporting allied health educational programs or allied health personnel in either an academic or health care setting. Because of the impossibility of covering the large number of and wide variety of allied health professions and occupations, the recommended publications are focused primarily on the educational programs listed and recognized by the American Medical Association and other accrediting bodies. Books and journals are categorized by subject; the book list is followed by an author/editor index and the subject list of journals by an alphabetical title listing. Items suggested for initial purchase (169 books and 32 journals) are indicated by asterisks. To purchase the entire collection of books and journals (2002 subscriptions) would require an expenditure of about $36,744. The cost of only the asterisked items totals $14,465. PMID- 12568156 TI - Developing an "evidence-based medicine and use of the biomedical literature" component as a longitudinal theme of an outcomes-based medical school curriculum: year 1. AB - This paper describes the new outcomes-based curriculum at the University of Miami School of Medicine, a model curriculum for the first decade of the twenty-first century. The new curriculum has a strong emphasis on evidence-based medicine (EBM), implemented throughout its four years as a component of one of its longitudinal themes. The "EBM and Use of the Biomedical Literature" component, which begins at orientation, was developed and is implemented by the Louis Calder Memorial Library, the center of EBM focus and activity for the curriculum and other initiatives at the University of Miami/Jackson Memorial Medical Center. The authors are unaware of any published reports of library-centric EBM initiatives as part of a longitudinal theme of a four-year outcomes-based curriculum. Other innovations of the EBM component in the new curriculum to date include use of Blackboard and CATmaker software programs for self-paced, interactive educational opportunities. PMID- 12568157 TI - An author co-citation analysis of medical informatics. AB - OBJECTIVE: This study presents the results of an author co-citation analysis of the interdisciplinary field of medical informatics. METHODS: An author co citation analysis was conducted for the years 1994 to 1998, using the fifty most cited American College of Medical Informatics fellows as an author population. Co citation data were calculated for every author pair, and multivariate analyses were performed to ultimately show the relationships among all authors. A multidimensional map was created, wherein each author is represented as a point, and the proximity of these points reflects the relationships of authors as perceived by multiple citers. RESULTS AND CONCLUSION: The results from this analysis provide one perspective of the field of medical informatics and are used to suggest future research directions to address issues related to better understanding of communication and social networks in the field to inform better provision of information services. PMID- 12568158 TI - Tribal connections health information outreach: results, evaluation, and challenges. AB - In 1997, the National Library of Medicine (NLM), a component of the National Institutes of Health (NIH), initiated a program of intensified outreach to Native Americans, initially focusing on the Pacific Northwest in collaboration with the Pacific Northwest Regional Medical Library (PNRML). This initiative, known as the Tribal Connections Project, emphasized the establishment or strengthening of Internet connections at select Indian reservations and Alaska Native villages and related needs assessment and training. The hope was that these efforts would improve tribal access to health information available via the Internet and the Web. Phase I included sixteen tribal sites--eight in Washington, four in Alaska, two in Montana, and one each in Oregon and Idaho. Phase I results indicate that the project was successful in assessing local needs and building awareness of the Internet, forging new partnerships with and between the participating Indian reservations and Alaska Native villages and other organizations, making real improvements in the information technology (IT) infrastructure and Internet connectivity at fifteen of sixteen sites, and conducting training sessions with several hundred tribal participants across thirteen sites. Most importantly, the project demonstrated the key role of tribal community involvement and empowerment and contributed to development of an outreach evaluation field manual and the evolving concept of community-based outreach. The knowledge gained from Tribal Connections Project Phase I is helping refine and enhance subsequent NLM sponsored tribal connections and similar community outreach efforts. PMID- 12568160 TI - Cost analysis and student survey results of library support for distance education. AB - This paper describes the costs associated with providing library support for a series of distance-education courses at The Ohio State University (OSU). These courses are designed as a pilot program offered by the OSU Office of Geriatrics and Gerontology. Costs to the library are analyzed for document delivery, electronic reserves, reference services, and librarian activities. Also included are the results of a student evaluation survey. The students are full-time working professionals who cannot attend regularly scheduled classes on campus. Conclusions extrapolate costs for each course, student, and service. PMID- 12568159 TI - General health problems of inner-city sex workers: a pilot study. AB - OBJECTIVE: A pilot study was designed to determine the general health problems of inner city sex workers. SAMPLE: The researchers worked with an agency that provides outreach services to these sex workers. Through this agency, they had access to a purposive sample of sex workers in a large Midwest city. METHODS: Nonparticipant observation was used to gather information about their health problems, the nature of information they may need, and the barriers to obtaining health care and health information. RESULTS: Sex workers (N = 75) ranged in age from nineteen to sixty-one years old. They identified a number of physical or psychological problems, such as rape, depression, and tuberculosis. HIV/AIDS was never mentioned. A major barrier to health care is a lack of information about where to go for treatment or how to obtain health insurance. CONCLUSIONS: More research needs to be done by library and information science professionals to determine the information needs of sex workers and the agencies that provide them with health and social services. PMID- 12568161 TI - Staff-driven strategic planning: learning from the past, embracing the future. PMID- 12568162 TI - The Association of Vision Science Librarians' citation analysis of Duane's Clinical Ophthalmology. PMID- 12568163 TI - Utilizing the past in the present curriculum: historical collections and anatomy at the University of Alabama School of Medicine. PMID- 12568164 TI - Proceedings, 102d Annual Meeting Medical Library Association, Inc. Dallas, Texas May 17-23, 2002. PMID- 12568165 TI - Amount of air infused to patient increases as fluid flow rates decrease when using the Hotline HL-90 fluid warmer. AB - OBJECTIVE: The intraoperative use of fluid warming devices has been recommended to avoid perioperative hypothermia and related adverse outcomes. To evaluate whether these devices might introduce risks of their own, we measured the volume of air escaping from a warmed intravenous solution that might be delivered to a patient. METHODS: In an operating room maintained at 19-19.5 degrees C, we tested an HL-90 Hotline fluid warmer with the L-70 fluid-warming set. One liter of lactated Ringer's solution was infused at flow rates of 150, 300, 500 and 3400 ml/h. The air that formed within the L-70 tubing during infusion was collected in a bubble trap placed at the end of the L-70 tubing. The volume of air in the bubble trap was measured. Twelve separate measurements were obtained at each flow rate. One additional study (n = 8) was performed using the L-10 Gas Vent to determine whether this equipment might reduce the volume of air infused when fluid flow rate was 300 mL/h. The volume of air collected at each flow rate was compared using ANOVA. RESULTS: Volume of air increased significantly from 1.0 +/- 0.2 mL to 2.9 +/- 0.4 ml as flow rate decreased from 3400 ml/h to 150 ml/h (p < 0.0001). The L-10 gas eliminator was ineffective in reducing the amount of air infused. CONCLUSIONS: We conclude that the use of the Hotline fluid warmer can result in infusion of air into the patient, introducing possible risk of air embolism. PMID- 12568166 TI - Optimal sequencing of urgent surgical cases. Scheduling cases using operating room information systems. AB - Optimal sequencing of urgent cases (i.e., selecting which urgent case should be performed first and which second) may enhance patient safety, increase patient satisfaction with timeliness of surgery, and minimize surgeons' complaints. Before determining the optimal sequence of urgent cases, an operating room (OR) suite must identify the primary scheduling objective to be satisfied when prioritizing pending urgent cases. These scheduling objectives may include: 1) perform the cases in the sequence that minimizes the average length of time each surgeon and patient waits; 2) perform the cases in the order that they were submitted; or 3) perform the cases based on medical priority, as prioritized by an OR director, or surgeons discussing the cases among themselves. We provide mathematical structure which can be used to program a computerized surgical services information system to assist in optimizing the sequence of urgent cases. We use an example to illustrate that the optimal sequence varies depending on the scheduling objective chosen. PMID- 12568167 TI - Implementation of a respiratory drive monitor on a Servo Ventilator. AB - OBJECTIVE: To design and evaluate a clinical monitor of respiratory drive (P0.1) and other respiratory variables in a simple way, using a commercial ventilator. METHODS: Nine healthy males were studied as they were breathing spontaneously in a Servo 900C Ventilator, at rest and during light exercise (50 W). The ventilator was slightly modified to improve its mechanical performance during spontaneous breathing, and was used as a measuring instrument. All the relevant information was retrieved, calculated and monitored by a PC. Respiratory drive was assessed as occlusion pressures from the inspiratory airway pressure signal. The equipment was compared with a two-way non-rebreathing laboratory system. Furthermore, negative and positive inspiratory pressures were applied from the ventilator, to study respiratory responses to mechanical loads. RESULTS: At rest, the ventilator introduced a minor influence on inspiratory time and P0.1, but not in ventilation, tidal volume, expiratory duration and respiratory frequency. During exercise, the influence was more evident. This effect could also be noticed in the coefficients of variation. The responses to mechanical loads were easily recorded and can be used as a simple test of central load-compensating mechanisms. CONCLUSIONS. The ventilator, with limitations, may be an alternative to conventional techniques, especially in clinical studies of the central inspiratory activity with and without respiratory loading. PMID- 12568168 TI - Evaluation of the HB-Quick: a portable hemoglobinometer. AB - OBJECTIVE: The Hb-Quick is a new portable hemoglobinometer that uses disposable cuvettes to measure the total hemoglobin concentration of capillary, venous, or arterial blood. Therefore, the objectives of this study were 1) to evaluate the performance of this compact, battery-powered hemoglobinometer by assessing its precision, accuracy, and linearity, 2) to determine whether its measurements suffer from interference by hemolysis, bilirubin, fetal hemoglobin, or hemodilution, and 3) to establish whether it can easily be used by clinical personnel with little or no laboratory training. METHODS: The precision of the test instrument was assessed by making repeated measurements on blood samples. Its accuracy and linearity were evaluated by comparing its measurements with the internationally accepted cyanmethemoglobin method. Samples of whole blood with and without bilirubin, fetal hemoglobin, hemolysis, and hemodilution were also analyzed to determine if any interference occurred when these disturbances were present, and it was placed in physicians' offices to evaluate its use by non laboratory personnel. RESULTS: Repeated measurements on blood samples with a wide range of hemoglobin concentrations were consistent with the precision specification (0.25 g/dl). The bias of the new hemoglobinometer was calculated as the mean difference between its readings and measurements with the cyanmethemoglobin method, and its accuracy as the standard deviation of the differences between the two methods. As assessed in this manner, the new hemoglobinometer had a bias of -0.04 g/dl and an accuracy of 0.40 g/dl. The linearity was checked over a hemoglobin concentration range from 0 to nearly 30 g/dl. There was a highly significant linear relationship between its readings and measurements with the cyanmethemoglobin method (slope = 0.997, y-intercept = 0.005, r = 0.999). Complete hemolysis of the sample increased the readings on average by only 0.22 g/dl. Bilirubin (17.5 mg/dl) increased the reading by an average of 0.29 g/dl, and fetal hemoglobin (76.5% HbF) reduced the readings by an average of only 0.035 g/dl. Diluting blood samples with saline also did not appreciably affect its accuracy. CONCLUSIONS: The new hemoglobinometer is fast and easy to operate. No sample preparation or pipetting is required. To operate the instrument, the user simply allows a drop of blood to fill the disposable cuvette by capillary action and inserts the cuvette into the instrument. The instrument analyzes the 10 microl sample and displays the results in less than 10 seconds. The interference caused by hemolysis, hemodilution with saline, fetal hemoglobin, and bilirubin were too small to be of any dinical importance. Tests in physician's offices indicated that clinical personnel with little or no formal laboratory training could successfully use this device. The observed precision, accuracy, linearity, and freedom from interference indicate that this hemoglobinometer is suitable for near-patient testing in a wide range of clinical settings including physicians' offices. PMID- 12568169 TI - Inter observer variability of the transcranial Doppler ultrasound technique: impact of lack of practice on the accuracy of measurement. AB - OBJECTIVE: Published data on the observer variability with the transcranial doppler (TCD) ultrasound are limited by the use of the product moment correlation coefficient. This study was designed to quantify the intra and inter observer variability with the TCD in terms of the intra class correlation coefficient (ICC) and to assess the impact of lack of practice on the observer variability and the accuracy of data generated. METHODS: The study was performed in two phases. In phase I, three observers insonated the middle cerebral artery in 10 healthy volunteers and measured peak systolic and end diastolic cerebral blood flow velocities. In phase II, the same observers repeated the experiment on five healthy volunteers after an eight week break during which none of the observers were allowed to use the TCD system. The observers were blinded to the measurements obtained. RESULTS: The ICC for peak velocity measurement was 0.9 (0.81-0.99) in phase I and 0.85 (0.66-1.00) in phase II (p < 0.05). The ICCs for end diastolic velocity measurements for phase I and II were 0.8 (0.64-0.96) and 0.67 (0.33-1.00) respectively (p < 0.01). CONCLUSIONS: A high level of observer agreement is possible with the TCD for measurement of cerebral blood flow velocities. Lack of regular practice with the system reduces the accuracy of measurements and impacts on both measured and calculated indices. PMID- 12568170 TI - Transcranial Doppler monitoring compared with invasive monitoring of intracranial pressure during acute intracranial hypertension. AB - OBJECTIVE: To determine whether a simple transcanial Doppler waveform variable pulsatility difference (systolic - diastolic blood flow velocity) can serve as a measure of critical changes in cerebral perfusion. METHODS: Thirteen pigs were anesthetized (anesthesia maintained with halothane) and ventilated to maintain normoxia and normocarbia. To measure mean arterial pressure, hemoglobin, and blood gases, the right carotid artery was cannulated. The right intracranial lateral ventricle was cannulated to measure and increase intracranial pressure; the right internal jugular vein was cannulated in 8 of 13 pigs to measure jugular venous oxygen saturation and to calculate cerebral arteriovenous oxygen content difference. Intracranial pressure was also monitored continuously with a subdural bolt in the contralateral frontal region, and blood flow velocity in the middle cerebral artery was measured with a transcranial Doppler probe on the right orbital region. Intracranial pressure was increased in increments of 10 to 20 mmHg by infusing saline through the ventriculostomy catheter until the transcranial Doppler indicated that blood flow velocity had ceased, at which point all variables were allowed to return to baseline. If mean arterial pressure failed to return to baseline, epinephrine, 0.01 to 0.1 microg/kg/min, was infused. Useful data were obtained from 8 pigs and were analyzed separately for pigs that received epinephrine (n = 4) and those that did not (n = 4). RESULTS: Transcranial Doppler measurements correlated more closely with cerebral perfusion pressure = (mean arterial pressure - intracranial pressure) than with intracranial pressure. In the range of 30 to 60 mmHg, cerebral perfusion pressure correlated linearly with the pulsatility difference. The closest nonlinear correlation (third order polynomial relationship) was noted between cerebral perfusion pressure and pulsatility difference (r = 0.8, P < 0.001, n = 217), for the animals that did not receive epinephrine. When a cerebral perfusion pressure < 60 mmHg and a cerebral arteriovenous oxygen content difference > 6.5 vol% were used to define limits of abnormal, pulsatility difference was a sensitive and specific indicator of abnormality in either variable. Pulsatility difference of > 70 cm/sec had > 77.1% and 86.7% positive accuracy rate, and < 0% and 14.3% negative accuracy rate for abnormal cerebral perfusion pressure (CPP) and cerebral arterio-venous O2 (C[a-v]O2), respectively. CONCLUSIONS: In pigs with induced diffuse intracranial hypertension, noninvasive transcranial Doppler waveform monitoring of pulsatility difference can identify increased cerebral oxygen extraction and dangerously decreased cerebral perfusion pressure. PMID- 12568171 TI - Point-of-care and standard laboratory coagulation testing during cardiovascular surgery: balancing reliability and timeliness. AB - OBJECTIVE: The use of point-of-care technology has increased faster than efforts to validate its effectiveness compared to standard laboratory testing modalities. To address this issue with a current point-of-care coagulation system (HEMOCHRON Jr, International Technidyne Corporation (ITC), Edison, NJ), we designed a study to test the hypothesis that data obtained from point-of-care coagulation equipment correlates with data obtained from standard laboratory coagulation equipment. One of the potential advantages gained using point-of-care testing is the ability to obtain more rapid results. To address this issue, turnaround time, defined as the elapsed time (in minutes) from when the sample was acquired from the patient until the investigators knew the results, was also determined. METHODS: Following Human Investigation Committee approval and informed consent, a prospective study was conducted to compare results obtained from point-of-care coagulation equipment with those results obtained from standard laboratory coagulation equipment. The study was performed in three groups of patients undergoing cardiovascular surgery, each requiring different levels of anticoagulation. RESULTS: Of the 83 patients who met the inclusion criteria, the correlation (combining data from groups 1-3) between results obtained from point of-care and standard laboratory prothrombin time was r = 0.867, p < 0.001. The correlation (group 3) between point-of-care and standard laboratory international normalized ratio was r = 0.943, p < 0.001. The correlation (combining data from groups 1 & 2) between point-of-care and standard laboratory activated partial thromboplastin time was r = 0.825, p < 0.001. Median turnaround time for the standard laboratory was 90 minutes, with a mean turnaround time of 74 to 78 minutes, depending upon the group. In contrast, the median turnaround time for point-of-care testing was two minutes and 14 seconds. CONCLUSIONS: The results from this study population reveal that data obtained from point-of-care prothrombin time, international normalized ratio and activated partial thromboplastin time results correlate with results obtained from standard laboratory coagulation testing. The value of obtaining reliable results in a timely fashion offers a potential advantage for point-of-care testing in dinical situations, such as in the operating room, where saving time may translate into financial savings. PMID- 12568172 TI - Assessment of short-term blood pressure variability in anesthetized children: a comparative study between intraarterial and finger blood pressure. AB - OBJECTIVE: Continuous blood pressure (BP) measurement provides instantaneous information on hemodynamic status, and allows for assessment of sympathetic modulation of vasomotor tone using spectral analysis. As an alternative to intraarterial blood pressure (IABP) measurement, the Finapres, a photoplethysmographic device, allows for non-invasive continuous measurement of finger blood pressure (FBP). This study was designed to evaluate the accuracy of spectral measurements of FBP variability in children during anesthesia and recovery. For this purpose, reliability of BP measurement and short-term BP variability assessed by FBP were calculated and compared with IABP. METHODS: Finger blood pressure was compared with IABP from the ipsilateral radial artery, in 14 children undergoing major surgery. Sixty-seven simultaneous recordings of both signals were performed during anesthesia and 32 during recovery period. The accuracy of the FBP was determined by measuring its bias and precision according to the Bland and Altman method. To assess the ability of the FBP to follow short term BP variability, bias of total spectral power and bias of the 3 main spectral components (LF, MF, HF) were calculated. Transfer functions between invasive and non-invasive signals were calculated. RESULTS: The average bias of SBP measurement was 3.8 +/- 7.4 mmHg during anesthesia and 2.2 +/- 6.7 mmHg during recovery. During anesthesia overall variability and spectral components of FBP and IABP were similar with both techniques; while during recovery, a selective amplification of the low frequencies (< 0.15 Hz) of FBP was observed. Frequency response analysis of the pressure waveform, showed a high coherence between both signal with a gain of 0.96 +/- 0.52 mmHg FBP/ mmHg IABP under anesthesia, and of 0.74 mmHg FBP/ mmHg IABP during recovery. CONCLUSIONS: The differences evidenced between FBP and IABP spectral profiles might result from specific physiological properties of digital arteries, which are sympathetic effectors. This study supports the use of FBP in children to assess non-invasively the vascular sympathetic component of the autonomic nervous system during anesthesia and recovery. PMID- 12568173 TI - On the dynamic performance of the Abbott Safeset blood-conserving arterial line system. AB - OBJECTIVE: Critically ill patients frequently have indwelling arterial lines placed during their Intensive Care Unit stay. The lines are used to monitor blood pressure continuously, administer drugs and to draw blood for a variety of physiologic tests. Several blood-conserving arterial line systems have been developed to eliminate the need to discard blood in the process of obtaining undiluted and uncontaminated blood samples. The purpose of this study was to evaluate the dynamic performance of one such system the Abbott Clinical Care System Safeset blood conserving arterial line system - in comparison to a conventional arterial line system. METHODS: We studied ninety-nine patients who had indwelling arterial lines placed during surgery and who were admitted to our Surgical Intensive Care Unit (SICU). The patients were randomly placed into one of two groups. The control group received a conventional indwelling arterial line system; the experimental group received the Abbott Safeset system. We measured the damping coefficient and resonant frequency daily in order to evaluate and compare the dynamic performance of the two systems. We also measured discard volumes (in the control group) and blood sample sizes during the patients' stays in the SICU. RESULTS: The two patient groups were similar in regards to demographics and baseline clinical characteristics. A median 3 ml of blood per draw and 17.5 ml of blood per patient was discarded in purging the conventional arterial line system while, by design, no blood was discarded with the experimental system. There was no difference between the two groups with regard to damping coefficient. Both systems were underdamped. However, the conventional arterial line system had a significantly higher resonant frequency (16.7 Hz) compared to the Safeset system (12.5 Hz). CONCLUSIONS: Because the Abbott Safeset blood-conserving arterial line system is underdamped and has a lower resonant frequency compared to the traditional arterial system, it may overestimate systolic blood pressure, particularly in patients with high heart rates. PMID- 12568174 TI - The clinical use of an esophageal Doppler monitor for hemodynamic monitoring in sepsis. PMID- 12568175 TI - Problem-based learning in residency education: a novel implementation using a simulator. AB - BACKGROUND: We developed a problem-based learning exercise with a full-scale human patient simulator to teach residents the emergency management and differential diagnosis of acute intraoperative hypotension. METHODS: We developed the exercise through the following steps: clear definition of learning objectives, preparation of an appropriate case stem, development of clinically realistic scenarios to illustrate objectives, and an interactive instructor to stimulate discussion. RESULTS: The exercise focused on the differential diagnosis of intraoperative hypotension, and the acute treatment of hypovolemia, cardiac tamponade, tension pneumothorax, and anaphylaxis. CONCLUSIONS: Exercises on a full-scale patient simulator are a natural extension of problem-based learning. Recent research in learning theory provides the rationale for this teaching modality's potential as a learning tool. PMID- 12568176 TI - Anorexia nervosa following bariatric surgery. PMID- 12568177 TI - A bariatric surgery algorithm. AB - BACKGROUND: Three premises underlie this formulation of an algorithm: 1) there is no gold standard operation; 2) the bariatric surgeon should be able to perform more than one bariatric operation; and 3) a patient can be broadly matched to an operation. METHODS: Literature review and interpretation. RESULTS: The 5 currently clinically-tested bariatric procedures, ranked from least to most weight loss, are: 1) gastric banding (GB); 2) vertical banded gastroplasty (VBG); 3) Roux-en-Y gastric bypass (RYGBP); 4) biliopancreatic diversion (BPD) or duodenal switch (DS); and 5) long-limb Roux-en-Y gastric bypass (LLRYGBP). The following diagram for decision-making takes into consideration body mass index (BMI); age; gender, race, and body habitus (GRH); and comorbidities (CoM): This diagram can be converted to an equation; OC = 1.0+BMI Number (1 to 6) +/- 0.5 (age <40>) +/- 0.5 (GRH, Favorable or Unfavorable) +/- 1 (CoM, Low or High), where OC = operative category: GB = 0 to 3, VBG = 2 to 5, RYGBP = 3 to 6, BPD/DS = 4 to 7, and LLRYGBP = 6 to 9. Overlap between OCs integers allows for surgeon and patient preference. CONCLUSIONS: A diagram or equation algorithm for operative selection in the morbidly obese has been constructed. PMID- 12568178 TI - Positioning the morbidly obese patient for anesthesia. AB - Each of the different positions employed during surgery can compromise cardiopulmonary function in the morbidly obese patient. An understanding of the physiologic changes that can occur is essential for the successful management of these patients. PMID- 12568179 TI - Pilot study examining the frequency of several gene polymorphisms in a morbidly obese population. AB - BACKGROUND: Obesity is a growing problem and is associated with numerous medical conditions. Several polymorphisms have been associated with lipid metabolism and obesity: PPAR-gamma2-Pro115Gln, PPARgamma2-Pro12Ala, beta3AR-Trp64Arg and SR-BI IVS5 C > T. We examined the frequency of these polymorphisms in patients with a BMI > 40 and compared them to individuals with a BMI < 30. Our hypothesis was that these polymorphisms would occur more frequently in the obese population. METHODS: This case-control study examined 126 individuals with a BMI > 40 in the McGill University Health Centre Bariatric Surgery Program and 102 individuals (controls) with a BMI < 30 attending a Lipid Clinic. DNA was extracted from whole blood by standard techniques. The polymorphisms were determined by polymerase chain reaction (PCR) and restriction genotyping. RESULTS: A significant difference between controls and the morbidly obese group was observed for 2 of 4 polymorphisms. The carrier frequency for PPARgamma2-Pro12Ala was 24.8% in the obese group and 12.9% in controls (odds ratio = 2.2; 95% CI = 1.1-4.4; P = 0.02). The carrier frequency for SR-BI IVS5 C > T was 22.8% in obese individuals versus 8.1% in controls (odds ratio = 3.5; 95% CI = 1.6-7.7; P = 0.002). There were no differences in frequency of diabetes in both obese (13/104) and control (9/126) groups (odds ratio = 1.86, 95% CI = 0.76 < O.R. < 4.54, P = 0.184). CONCLUSIONS: These results underscore the relationship between gene polymorphisms and obesity. Obese individuals may differ from non-obese individuals in the gene polymorphisms associated with metabolic control. PMID- 12568180 TI - The effect of erythromycin on bile excretion and proximal small bowel motility following divided gastric bypass surgery: a prospective randomized placebo controlled trial. AB - BACKGROUND: No conclusive data exists supporting the use of any prokinetic agent in the postoperative setting. The study was designed to examine the effect of erythromycin on small bowel motility in a placebo-controlled trial of post gastric bypass patients utilizing a standardized nuclear medicine test. METHODS: A consecutive series of 21 patients undergoing elective gastric bypass surgery for morbid obesity between September 1999 and March 2001 were enrolled in this prospective double-blind randomized controlled trial. Standard open, divided gastric bypass was performed. Patients were randomized to receive either erythromycin 250 mg i. v. (11 patients) or placebo (10 patients) every 8 hours. On postoperative day 2, a hepatic iminodiacetic acid (HIDA) scan was obtained. Tracer movement through the biliary tree and proximal small bowel was quantified and compared. RESULTS: Tracer clearance from the liver and biliary tree was no different between groups from time of injection through 1 hour. Tracer material clearance from the duodenum into the jejunum was no different between the erythromycin and control groups at 1 hour, 37% +/- 13% and 37% +/- 22% respectively (P = 0.95). At 4 hours, clearance was greater in the erythromycin group, 77% +/- 6%, compared to control, 60% +/- 20% (P = 0.036). The rate of tracer change between hour 1 and 4 (slope) was steeper in the erythromycin group (P = 0.048). CONCLUSIONS: Erythromycin increases intestinal transit in the postoperative setting. PMID- 12568181 TI - Health-related quality of life in morbidly obese patients: effect of gastric bypass surgery. AB - BACKGROUND: The effect of gastric bypass on the health-related quality of life (HRQoL) of morbidly obese patients was investigated in a cross-sectional study. METHODS: A postoperative group of 78 patients on average 13.8 years after gastric bypass was compared with a preoperative control group of 110 patients. The SF-36 was used to assess HRQoL. In preoperative patients, the SF-36 was self administered, while in the postoperative group, telephone interviews were conducted. In the postoperative sample, multiple stepwise linear regression analyses were carried out to examine putative predictors of the physical (PCS) and the mental (MCS) composite scores of the SF-36. RESULTS: Significant differences between the pre-and postoperative group were found for all subscales except Mental Health, in favor of the postoperative group. On average 13.8 years after gastric bypass, most of the sub-scales were similar to the US norm values. However, the Bodily Pain and the overall Physical Composite scale (PCS) scores were lower (more impaired) in the postoperative group compared with the US norms. Female patients, patients who were hospitalized since the surgery, and those who had lost less weight had more impaired values on the PCS and patients who reported binge-eating disorder (BED) at follow-up had more impaired values on the Mental Composite Scale (MCS) of the SF-36. CONCLUSION: HRQoL was significantly better in postoperative gastric bypass patients in comparison to a sample of preoperative patients. However, HRQoL, specifically the physical domain of the SF 36, was more impaired in long-term follow-up patients compared with US norm values. The reoccurrence of BED after surgery negatively influenced the mental domain of the SF-36. PMID- 12568182 TI - Perceived psychosocial outcomes of gastric bypass surgery: a qualitative study. AB - BACKGROUND: Attempts to understand postoperative psychosocial changes in the lives of individuals who have undergone gastric bypass surgery for morbid obesity have 1) been guided by constructs emanating from the assumptions of researchers, and 2) have resulted in fragmented conclusions that catalogue changes without theoretically integrating them. MATERIALS AND METHODS: Using unstructured and semi-structured interviews and in-depth focus groups, 31 patients were asked in an open-ended fashion about the ways, if any, in which gastric bypass surgery had affected their lives. Grounded theory methodology was utilized in order to identify emergent themes and their interrelations, and build a meaningful, comprehensive theory of life after gastric bypass. RESULTS: Patients' report of a rebirth/transformation was identified as the core process of the theory. The changes marking this process were clearly conceptualized in dichotomous terms comparing pre to postsurgical life. Patients reported changes that they regarded as unequivocally positive, a number of which had not been previously reported in the literature. Unique to this particular study was the finding of numerous life changes that generated tension and posed challenges in various aspects of patients' lives. CONCLUSION: The grounded theory proposes that the extent to which patients successfully negotiate tension-generating changes may be a major determinant in the long-term outcome of gastric bypass, both weight loss and psychosocial adjustment. PMID- 12568183 TI - Sweet eating is not a predictor of outcome after Lap-Band placement. Can we finally bury the myth? AB - BACKGROUND: It is common belief that sweet eaters will do poorly after gastric restrictive surgery. There is scant evidence for this and significant evidence that sweet eating behavior is not predictive of weight outcome. Preoperative and current sweet eating behavior was assessed in subjects who have had Lap-Band surgery, to find if this influenced weight outcomes. METHOD: 200 unselected patients who had bands inserted for > 1 year completed a questionaire regarding preoperative sweet eating behavior. The last 100 patients also reported current sweet eating behavior. Sweet eating was scored using a standard dietary questionnaire. RESULTS: Mean +/- SD % excess weight loss at 1 year (% EWL1) for the 100 with the highest preoperative sweet eating scores (SES) was 47.1 +/- 16% compared with a loss of 48.2 +/- 16% by those with the lowest SES (P = 0.64). Analysis showed no significant linear or non-linear correlation between the SES and the % EWL. For the highest quintile of SES, the EWL1 was 47.3 +/- 14% and for the lowest was 46.1 +/- 16% (NS). Sweet eaters were younger (r = -0.21, P = 0.003) and had higher fasting insulin concentrations (r = -0.18, P = 0.03). Preoperative SES had no influence on % EWL1 after controlling for factors known to influence weight loss. % EWL at 2 years (n = 130) and 3 years (n = 88) were not different for sweet eaters and non-sweet eaters. Current sweet eating tendency (n = 100) also had no impact on % EWL. CONCLUSION: Sweet eaters do not have less favorable weight outcomes following Lap-Band surgery. Our study confirms the findings of two other major studies. Sweet eating behavior should not be used as a preoperative selection criterion for bariatric surgery. PMID- 12568185 TI - Liver damage in obese patients. AB - BACKGROUND: Hepatic steatosis, nonalcoholic steatohepatitis and cryptogenetic cirrhosis are frequent in an obese population. Therefore, it is wise to submit all obese patients with significant alterations in hepatic function tests to transparietal liver biopsy. The aim of this study is to determine the hepatic conditions of morbidly obese patients during bariatric surgery by means of a wedge liver biopsy, to avoid any eventual hepatic damage being ascribed to the surgical procedure. METHODS: This prospective study entails 216 consecutive patients, whose work-up included liver function tests, before undergoing vertical gastroplasty and wedge liver biopsy. Histology was assessed for hepatic steatosis, necroinflammatory activity and liver fibrosis/cirrhosis. RESULTS: Abnormal preoperative liver function tests were detected in 65 patients, in 52 unexpectedly. Histologically, significant steatosis was found in 168 patients (77.8%); necroinflammatory activity in 13 (6.0%); liver fibrosis in 46 (21.3%), 5 of whom (2.3%) were found to have an asymptomatic and unknown liver cirrhosis. CONCLUSION: In morbidly obese patients, the incidence of histological liver damage is very high, despite acceptable liver function tests. In addition to steatosis, however, a "second hit" to induce necrosis and inflammation, favoring the development of significant fibrosis, is not essential. Being obese is an independent risk factor for liver damage and could contribute to liver fibrosis either alone or in association with other insulting factors. The identification of obese patients with septal fibrosis/cirrhosis, at surgery, is of considerable interest in clinical practice, mainly under the aspect of prognosis and liability. PMID- 12568184 TI - Effect of surgical weight loss on free radical and antioxidant balance: a preliminary report. AB - BACKGROUND: This study observes the effect of surgical weight loss on free radical and antioxidant vitamin balance. PATIENTS AND METHODS: 22 consecutive morbidly obese patients undergoing vertical banded gastroplasty (VBG) were chosen for the study. Postoperative studies were done at 12 and 24 weeks. Plasma antioxidant and vitamin determinations were performed by HPLC method. RESULTS: Subjects lost a significant amount of weight (P < 0.01). Compared to preoperative measurements, postoperative measurements of plasma beta-carotene were not statististically different both at 12 and 24 weeks (13.86 +/- 1.26 microg/dl, 12.35 +/- 1.2, P = 0.44; 14.33 +/- 2.03, P = 0.77; preoperatively, 12 and 24 weeks respectively). Alpha-tocopherol increased slightly at the 12th week; the difference was not significant (8.50 +/- 0.77; 9.56 +/- 0.82, P = 0.37; preoperatively and 12th week respectively). The levels of alpha-tocopherol rose at 24th week significantly (10.89 +/- 0.55, P = 0.028). The indicator of lipid peroxidation (malondialdehyde) decreased with weight loss (1.505 +/- 0.11 micromol/L preoperatively; 0.75 +/- 0.062 at 12th week, P = 0.01; 0.712 +/- 0.05 at 24th week, P < 0.01). CONCLUSION: Our data show that free radical generation falls markedly in association with weight loss after VBG. Surgical weight loss leads to significant decrease in oxidant production and also leads to increase in some antioxidant vitamins. The demonstration of decreased free radical generation and correction of balance between free radicals and antioxidant vitamins has important implications for oxidative mechanisms underlying obesity-associated disorders. PMID- 12568186 TI - Long-term dietary changes after vertical banded gastroplasty: is the trade-off favorable? AB - BACKGROUND: Insufficient data exist about the long-term health consequences of gastric restriction procedures used for treatment of obesity. The long-term nutritional changes that occur after vertical banded gastroplasty (VBG) were evaluated. METHODS: All consecutive patients who underwent VBG surgery in one surgical ward were invited for a follow-up study 3-10 years after the surgery. Demographic and clinical characteristics were evaluated from the patients' medical charts. Dietary assessment was performed using a food frequency questionnaire, which included 52 frequently consumed food items. RESULTS: Of the 122 patients who underwent VBG between 1986-1992, 75 (62%) participated in the follow-up study. The average time since surgery was 5.4 +/- 1.8 years, and the average weight loss was 24.9 +/- 12.4%. Most of the patients eat only one major meal daily, and only one-third regularly ingest solid foods. Dietary analysis revealed a decreased intake of most nutrients compared with pre-surgery, with the exception of dairy products, sweet foods and fluids. The greatest decrease was found in the consumption of fiber-rich fruits and vegetables, followed by meat, fish and complex carbohydrates. CONCLUSION: While the weight loss itself and the reduction in fat consumption that are seen after VBG are probably beneficial, the long-term effects of the decreased consumption of fruit, vegetables, other complex carbohydrates and fish may counterbalance these benefits. The net effect of this trade-off on future health is difficult to predict and requires long-term evaluation of clinical outcome. PMID- 12568187 TI - Open versus laparoscopic vertical banded gastroplasty: a randomized controlled double blind trial. AB - BACKGROUND: Vertical banded gastroplasty (VBG) is a frequently used surgical procedure for the treatment of morbid obesity. It can be done open (OVBG) or laparoscopic (LVBG). The aim of this double-blind randomized clinical trial was to compare the postoperative outcome and 1-year follow-up of 2 cohorts of patients who underwent either OVBG or LVBG. PATIENTS AND METHODS: 30 patients with morbid obesity were randomized into 2 groups (14 OVBG and 16 LVBG). Pain intensity, analgesic requirements, respiratory function, and physical activity were blindly analyzed during the first 3 postoperative days. Complications, weight loss, and cosmetic results after 1 year follow-up were evaluated. RESULTS: Both groups were highly comparable before surgery. Surgical time was longer in the laparoscopic procedure. Patients in this group required less analgesics during the first postoperative day. There was an earlier recovery in the expiratory and inspiratory forces, as well as faster recovery of physical activities in patients who underwent LVBG. Postoperative complications were more frequent in the open group. Excess body weight loss after 1 year was similar in both groups. Cosmetic results were significantly better in the laparoscopic group. CONCLUSIONS: LVBG had advantages over the open procedure in terms of analgesic requirements, respiratory function, postoperative recovery, and cosmetic results. PMID- 12568188 TI - Gastrointestinal quality of life following laparoscopic vertical banded gastroplasty. AB - BACKGROUND: Laparoscopic vertical banded gastroplasty (LVBG) is a safe and effective treatment for morbid obesity. Previous studies disclosed a significant improvement in the health-related quality of life after substantial weight loss following VBG. Data regarding the specific gastrointestinal quality of life following LVBG is lacking. MATERIALS AND METHODS: 223 patients who underwent LVBG for morbid obesity were studied prospectively. Quality of life was measured by the Gastrointestinal Quality of life Index (GIQLI), a 36-item questionnaire before surgery, and at 6 months, 1 year and 2 years after surgery. The questionnaire is divided into 5 domains, and the maximum score is 144. RESULTS: After LVBG, weight loss has been good. Mean BMI decreased from 43.2 to 31.3 after 2 years. Co-morbidities were eliminated in 71%. 84.3% of patients were satisfied with the results. However, the score of GIQLI remained similar before and after surgery. Preoperative score was 106.2 +/- 19 points. The score became 116.6 +/- 9, 106.8 +/- 21, and 108.5 +/- 20 at 6 months, 1 year and 2 years after surgery respectively. The patients had improvement in 3 domains of the questionnaire (social function, physical status and psychological emotions) but decreased in domains of core symptoms and disease-specific items. CONCLUSION: Although LVBG was effective in reduction of weight and resolution of co-morbidities in morbidly obese patients, the specific gastrointestinal quality of life did not improve. Many patients developed some specific gastrointestinal symptoms in order to obtain weight reduction. PMID- 12568189 TI - Five-year results of laparoscopic vertical banded gastroplasty in the treatment of massive obesity. AB - BACKGROUND: Laparoscopic surgery appears to offer rapid recovery and low postoperative morbidity. The aim of the present study was to assess the outcome of laparoscopic vertical banded gastroplasty (LVBG) in 154 obese patients with a follow-up of 12-60 months. PATIENTS AND METHODS: 154 massively obese patients (132 female) with a mean +/- SEM body mass index (BMI) of 43.4 +/- 0.6 kg/m2 were followed prospectively for an average of 31.7 +/- 1.4 months. LVBG was performed using 5 trocars placed in a standard fashion for laparoscopic upper gastrointestinal surgery. A 4-row stapler was used for the vertical staple-line and a stretched polytetrafluoroethylene (Gore-tex) band was used to reinforce the outlet. After the first 67 cases, the procedure was altered so that a 5-cm length was marked on the band. RESULTS: Conversion to open surgery was performed in 33 cases. All patients lost weight. At 60 months follow-up, the postoperative weight was similar in the open and laparoscopic group.The subjects where 5 cm length was marked on the band had a significantly better weight loss at 36 months (30.4 +/- 1.2). Both early (< 1 month postoperative) and late (> 1 month postoperative) complications were more common in the group converted to open surgery. Postoperative stay was shorter in the laparoscopic group. CONCLUSIONS: LVBG can be performed safely and results in shorter postoperative stay than open VBG. With adherence to surgical technique (5-cm band circumference), weight-loss is maintained at an adequate level. Complications after LVBG do not exceed open VBG. PMID- 12568190 TI - Medial thigh lift free flap for autologous breast augmentation after bariatric surgery. AB - BACKGROUND: Massive weight loss following bariatric surgery frequently results in body contour deformities like ptotic and hypoplastic breasts, redundant abdominal tissue and loose skin especially in the medial thigh area. This redundant tissue can be used for breast augmentation in the case of hypertrophic ptotic breasts. METHOD: In 3 patients who underwent a vertical banded gastroplasty and consecutively lost more than 60% of their body weight, a breast augmentation with a transverse gracilis myocutaneous free flap was performed. RESULTS: Bilateral myocutaneous gracilis free flap breast augmentation resulted in an esthetic, pleasing result, with additional correction of the redundant skin from the medial thigh region. CONCLUSION: Autologous breast augmentation with a simultaneous medial thigh lift can be performed safely, after successful weight loss following bariatric surgery. PMID- 12568191 TI - Psychopathological status and interpersonal functioning following weight loss in morbidly obese patients undergoing bariatric surgery. AB - BACKGROUND: We questioned whether differences in psychopathological status and interpersonal relations exist in a group of morbidly obese patients 18 months after bariatric surgery, as related to extent of weight loss. METHODS: The study group consisted of 100 morbidly obese patients (85 female, 15 male) who had undergone surgical treatment (vertical banded gastroplasty) for weight reduction. Each patient completed the Lancashire Quality of Life Profile European version, the Eating Disorder Inventory, the Symptom Check List-90-Revised, and the Millon Clinical Multiaxial Inventory-II. The sample was divided into 2 groups according to the percentage of excess weight loss 18 months after surgery: a greater weight loss group (weight loss > 30%) and a lesser weight loss group (weight loss < 30%). RESULTS: Significant differences were found between the 2 groups in percentage of weight loss (P < 0.0001), negative self-esteem (P < 0.001), drive for thinness (P < 0.001), body dissatisfaction (P < 0.001), global EDI (P < 0.002), anxiety (P < 0.003), GSI (P < 0.002), avoidant (P < 0.001), borderline (P < 0.0001), and passive-aggressive (P < 0.002). CONCLUSION: Greater weight loss strongly correlates with improved quality of life, less disturbed eating behavior, and lower psychopathology. These results justify the clinical use of surgical procedures and demonstrate that weight loss has powerful psychological and psychosocial implications. PMID- 12568192 TI - Weight loss after Swedish Adjustable Gastric Banding: relationships to insulin resistance and metabolic syndrome. AB - BACKGROUND: The metabolic syndrome is a cluster of cardiovascular risk factors (central obesity, hypertension, dyslipidemia, disturbance in glucose metabolism) associated with insulin-resistance. The cluster of risk factors defining the metabolic syndrome increases cardiovascular risk more than each single component. The aim of the present longitudinal study was to evaluate the relationship between weight loss and changes in insulin-resistance and in the prevalence of the metabolic syndrome 1-year after SAGB implantation. METHODS: 51 premenopausal severely obese women (mean age 35.2 +/- 8.8 years, BMI 43.3 +/- 6.9) were enrolled. As a control group, 51 premenopausal non-obese women (BMI < 30) were enrolled. All obese subjects underwent successful implantation of the SAGB via videolaparoscopy. In all subjects insulin-resistance was estimated by HOMA index and metabolic syndrome was defined according to the criteria of the European Group for the Study of Insulin Resistance. RESULTS: HOMA (4.2 +/- 2.0 vs 1.9 +/- 0.8, P < 0.001) and the prevalence of the metabolic syndrome (58.8% vs 7.8%, P < 0.001) were significantly higher in obese than non-obese women. 1 year after SAGB, BMI significantly decreased from 43.3 +/- 6.9 to 34.5 +/- 7.4 (P < 0.001). HOMA index showed a significant dramatic breakdown (4.2 +/- 2.0 vs 2.4 +/- 1.0, P < 0.001). The prevalence of the metabolic syndrome declined significantly (58.8% vs 21.6%, P < 0.001). CONCLUSION: Our study shows that in severely obese women, insulin-resistance and the prevalence of the metabolic syndrome significantly decrease 1 year after SAGB. Our findings indicate that SAGB could be a useful tool to reduce the global cardiovascular risk in severely obese people and to improve their long-term prognosis. PMID- 12568193 TI - Results of the Italian multicenter study on 239 super-obese patients treated by adjustable gastric banding. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) is the most common bariatric operation. This study is a retrospective analysis of the multicenter Italian experience in patients with BMI > 50 over the last 4 years. METHODS: An electronic data sheet made for LAGB-operated patients since January 1996, was mailed and e-mailed to all surgeons involved in this kind of procedure in Italy. Items regarding patients with BMI > 50 were selected. Analysis used Fisher's exact test and logarithmic regression analysis (P < 0.05 significant). Data were expressed as mean +/- SD. RESULTS: 239 patients (13.3%), out of 1,797 Lap-Band operated patients entered the study (179F / 60M), with mean age 37.6 +/- 11.3 years (19-69) and mean BMI 54.6 +/- 4.8 (50.1-83.6). Laparotomic conversion rate was 5.4% (44/239). Postoperative complications occurred in 24 / 239 patients (9.0%). Follow-up was obtained in 218 / 218, 198 / 198, 121 /147, 75 / 93, 30 / 38 LAGB patients at 6,12, 24, 36, and 48 months respectively. At these time periods, mean BMI was 46.7, 43.9, 42.2, 41.9, and 39.3 kg/m2. At the same intervals, mean %EWL was 24.1, 34.1, 38.8, 38.9, and 52.9%. The number of patients with < 25% EWL at 12, 24, 36, and 48 months follow-up were 34, 10, 4, and 0. Serious co-morbidities (189 in 124 of 239, 57%) had completely resolved 1 year postoperatively in 74 / 124 of the patients (59.6%). CONCLUSION: Although super-obese patients following the LAGB remain obese with BMI > 35, in the short term most lose their co-morbidities, with a very low morbidity and mortality rate. PMID- 12568194 TI - Re-operations following laparoscopic adjustable gastric banding. AB - BACKGROUND: Re-operations after laparoscopic adjustable gastric banding operation (LAGB) are band-associated or due to complications of the access-port. Symptoms, diagnostics, operations, and follow-up of patients with re-operations were analyzed. METHODS: Between December 1996 and January 2002, 250 morbidly obese patients were treated with LAGB and prospectively evaluated using a standardized protocol. Since June 2000 the pars flaccida technique was applied, since October 2000 with the new 11-cm Lap-Band. All adjustments of the band were done under radiological control. RESULTS: Of 250 patients, 39 had to be re-operated because of band-associated complications: 27 laparoscopic re-gastric bandings after 12 (3 26) months because of slippage; 6 laparoscopic removals of the band (band intolerance - 4, pain - 1, pouch dilatation - 1); 12 biliopancreatic diversions with duodenal switch (BPD-DS) after 29 (18-43) months due to pouch and/or esophageal motility disorders (9) or insufficient weight loss (3), in 6 patients after having already performed a re-banding for slippage. 9 revisions of the access-port were done after 6 (2-53) months (disconnection - 3, dislocation - 6). The morbidity of the re-operations was 5.3%: 1 hematoma in the abdominal wall and 1 temporary dysphagia after re-banding, 1 pulmonary embolism following BPD-DS. There have been no deaths. In patients with a minimal follow-up of 3 years (n = 92), the yearly re-operation rate was 11-12%. No slippage has occurred with the new 11-cm Lap-Band. CONCLUSION: Re-operations after LAGB for band-associated complications were frequent but could be performed safely with little morbidity. When the new 11-cm Lap-Band was employed, the high slippage rate dropped. PMID- 12568195 TI - Laparoscopic adjustable gastric banding: essential role of psychological support. AB - BACKGROUND: A follow-up study of laparoscopic adjustable gastric banding (LAGB) in morbidly obese patients was conducted, with special emphasis on psychological aspects. MATERIALS AND METHODS: 128 patients (98/30 females/males) underwent LAGB between October 1995 and March 2001. Age was 38 +/- 9 years, body mass index (BMI) 42.72 +/- 4.25 kg/m2 and body weight 124 +/- 25 kg (mean +/- SD). Weight loss, early and late complications, band calibrations and difference between patients with and without psychotherapy were analysed. Psychological status was investigated before and during LAGB treatment using MMPI. Quality of life was investigated using MOS SF36. RESULTS: Median BMI decreased from 42.7 preoperatively to 31.0 kg/m2 after 12 months. Reduction of BMI was already significant (P < 0.01) after 3 months. In the following 12 months, median BMI wavered between 31.0 and 29.5 kg/m2. Best results were achieved in patients who followed psychotherapy. CONCLUSION: LAGB gave therapeutic success (progressive and significant weight loss). Psychotherapy was not only a simple support to diet but also an important part of the multidiscipline treatment. For best results, the team must care for patients individually and not just follow the same fixed protocol. PMID- 12568196 TI - Coping behavior and loneliness among obese patients. AB - BACKGROUND: Morbid obesity can be accompanied by physical and social problems that may influence interpersonal relationships and the recruitment of social support. The problems can be tackled with a variety of coping strategies. METHODS: 104 patients with a body mass index (BMI) 32-64 kg/m2 and mean age 36 yr were presented with the Utrecht Coping List (UCL) and the Loneliness Scale. Of these patients, 94 were female, and this cohort was analyzed more extensively. RESULTS: Patients exhibited elevated values on the Loneliness Scale and in the UCL sub-scales palliative response, avoidance / wait-and-see, passive / depressive response pattern and expression of emotions / anger. The active approach UCL sub-scale scored lower than in a control group. CONCLUSION: Obese female patients displayed avoidance, wait-and-see and passive response pattern as coping behavior, experiencing their intimate relationships as relatively unreliable and not very intimate. More research is needed to determine the effect of coping behavior on therapeutic effect. PMID- 12568197 TI - Anthropometric measurements in male breast cancer. AB - BACKGROUND: 1% of breast cancers occur in men. The etiology is obscure. An elevated BMI has been postulated to be a cause. METHODS: All male breast cancer patients operated from January 1990 to May 2001 were retrospectively reviewed. Relation between BMI and male breast cancer was examined. RESULTS: 43 males underwent breast surgery for breast cancer during this period. 3 patients were excluded from the study because of other risk factors for breast cancer. The average BMI of 40 patients was 26.54 kg/m2, which is mildly above the level for normal weight. CONCLUSIONS: Excessive adipose tissue may increase risk of male breast cancer. PMID- 12568198 TI - Immobilization hypercalcemia in critical illness following bariatric surgery. AB - BACKGROUND: Immobilization hypercalcemia has been previously reported in a number of entities but not as a complication of bariatric surgery. We recognized this complication in two consecutive bariatric patients requiring intensive care unit (ICU) admission. METHODS: These two patients are reported in detail, and a review of our ICU database is also reported. RESULTS: Treatment of immobilization hypercalcemia in these two patients with pamidronate was successful. CONCLUSION: Immobilization hypercalcemia complicating the course of bariatric patients requiring ICU admission is a newly recognized and treatable entity. PMID- 12568199 TI - Carcinoid tumor and bariatric surgery. AB - Carcinoid is a rare gastrointestinal tumor. We have noticed a high incidence of carcinoid in our patient population during the gastric reduction-duodenal switch operation. Further studies are needed to identify any potential correlation between morbid obesity and carcinoid tumors. PMID- 12568200 TI - Functional gastric bypass with an adjustable gastric band. AB - BACKGROUND: To create a bond between the technique proposed and the patient, flexibility and adaptability are indispensable. Thus, we devised the functional gastric bypass, which can be activated and deactivated by inflating or deflating the Lap-Band. METHODS: The Lap-Band was positioned according to the standard technique, with the addition of a hand-sewn side-to-side gastroenterostomy between the gastric pouch and the intestine in the form of an Omega loop. Inflation or deflation of the Lap-Band allows activation or deactivation of the bypass. From October 1995 to December 2001, 545 Lap-Band operations were carried out. Between January 2001 and December 2001, functional gastric bypass was performed on 7 patients. Indications were: 1) prior failed Lap-Band treatment; 2) the first-choice operation for patients where non-restrictive surgery may be indicated. RESULTS: There was no morbidity or mortality. The functioning bypass was confirmed radiologically and clinically. CONCLUSION: In morbid obesity characterized by a variable relationship with food, the flexibility of the functional bypass allows adaptation to changes in the pathology itself and in the individual patient, which other surgical techniques cannot do. Surgical indications, proven feasibility, safety and efficacy await long-term documentation. PMID- 12568201 TI - Comments to presidential address: gastric bypass and biliopancreatic diversion operations. PMID- 12568202 TI - Homocysteine--are we making a mountain out of a molehill? PMID- 12568203 TI - Plasma homocysteine concentration and coronary artery disease in Asian Indians. AB - OBJECTIVE: This study was done (a) to evaluate the relationship between the plasma total homocysteine (tHcy) levels and coronary artery disease (CAD) in Asian Indians and (b) to see the relationship between tHcy and glucose intolerance. METHODS: Fasting concentrations of plasma tHcy was measured in 137 men, aged > or = 25 years who underwent coronary angiography while investigating for chest pain. Among them 71 had no CAD and 66 had CAD. Fasting plasma glucose (FPG) and glycosylated haemoglobin (HbA1c) were estimated. Total Hcy was measured using the Elisa method (Axis Biochemicals ASA-Oslo, Norway) in fasting EDTA plasma. RESULTS: The subjects with CAD were significantly older but had similar body mass index (BMI), waist-hip ratio (WHR), FPG and HbA1c values compared with the non-CAD subjects (P < 0.001). The median tHcy and the percentages of abnormal values were similar in non-CAD and CAD groups. No significant differences were seen in the four subgroups with respect to the mean tHcy or the percentage of abnormal values. The highest tHcy values were seen in the non-diabetic, non-CAD group (group 1--control). CONCLUSION: This preliminary data indicates that tHcy concentrations are not elevated in subjects with CAD and probably there is no association between total homocysteine and CAD in Indians. Homocysteine values were not influenced by the glucose tolerance status. Measurement of homocysteine concentrations may be more appropriate when the blood levels of vitamin B12 and folate are also measured. PMID- 12568204 TI - Immunoreactive insulin response to a single dose of glimepiride in lean type 2 diabetic subjects. AB - BACKGROUND: The best method for glucose lowering in lean type 2 diabetes remains controversial and this study was undertaken to study the 24 hour insulin response of these diabetics to glimepiride, a sulfonylurea with distinctive properties. METHODS: Twenty five consecutive newly diagnosed diet-unresponsive lean type 2 diabetics (BMI < 19 kg/m2) without any vascular complications were given single dose (1 mg) of glimepiride and insulin responses were measured 2,4,8,12 and 24 hours later. Pre and post-glimepiride blood glucose levels were also measured. RESULTS: All the post-glimepiride insulin levels were significantly higher than basal values. Increase in insulin secretion peaked at four hours and benefits lasted for at least 24 hours. This was accompanied by clinically and statistically significant reductions in fasting and postprandial blood glucose levels. Maximum secretory response correlated positively with beta cell function (HOMA) and negatively with fasting glucose. CONCLUSIONS: Glimepiride improved insulin secretion and hyperglycemia in lean type 2 diabetic subjects, with benefits lasting for 24 hours. The degree of response was proportional to the beta cell reserve, and occurred irrespective of the presence or absence of markers of insulin resistance. PMID- 12568205 TI - An open comparative clinical trial to assess the efficacy and safety of losartan versus enalapril in mild to moderate hypertension. AB - OBJECTIVES: To compare the efficacy and safety of losartan with enalapril, in mild to moderate hypertension. METHODS: An open, enalapril controlled study was conducted in 30 patients with mild to moderate hypertension. Losartan 50 mg was administered to patients for eights weeks. Throughout the study blood pressure was measured every two weeks. Routine laboratory investigations were performed before entering the trial, fourth week and at the end of the study. Adverse effects were recorded. After eight weeks losartan was stopped and enalapril 10 mg daily was administered to the same patients after two weeks washout period. The same methodology that was followed for losartan trial was repeated for enalapril trial also. RESULTS: Losartan treatment resulted in a highly significant reduction in the mean sitting diastolic blood pressure. Comparison with enalapril showed that both drugs are equally efficacious in reducing blood pressure in mild to moderate hypertension. The percentage of responders was slightly more with losartan than enalapril (86.7% vs 76.7%). Adverse events reported with losartan were mild. Enalapril also was well tolerated like losartan but there was high incidence of dry cough, which was reported in nine patients (30%). CONCLUSIONS: Losartan is an effective antihypertensive drug with an excellent safety and tolerability profile. It shows similar blood pressure lowering efficacy to that of enalapril. In contrast to enalapril, losartan does not cause dry cough. PMID- 12568206 TI - Comparison of colour duplex ultrasound and ankle-brachial pressure index measurements in peripheral vascular disease in type 2 diabetic patients with foot infections. AB - BACKGROUND AND OBJECTIVES: Peripheral vascular disease (PVD) is a common cause of morbidity and mortality and is reported to be more common among diabetic subjects compared to non-diabetic subjects. The aim of the study was to compare the specificity and sensitivity of ankle-brachial index (ABI) measured by peripheral doppler with the colour duplex ultrasound (CDU) for diagnosis of PVD. METHODS: One hundred type 2 diabetic patients admitted to our diabetic centre with foot lesions underwent both colour duplex ultrasound and ankle-brachial index measurements. PVD was diagnosed if the individual had haemodynamically significant obstruction on CDU, or if the ABI was < 0.9. The sensitivity and specificity of ABI was determined using the CDU as 'gold standard'. RESULTS: The mean age of the study group was 59.5 +/- 10.1 years and the mean duration of diabetes was 11.7 +/- 8.1 years. Of the total 100 subjects, six subjects had calcification of peripheral vessels and they were not included while calculating for sensitivity and specificity of ABI. Twenty (21.3%) subjects diagnosed as PVD by the CDU were not classified as PVD by the ABI measurements. Conversely, only three subjects (3.2%) classified as PVD by ABI had normal arteries based on CDU scanning. Overall, ABI had low sensitivity (70.6%) but a high specificity (88.5%). The overall agreement between CDU and ABI was poor (k = 0.20). CONCLUSION: ABI is a good initial screening tool but some patients with significant stenosis in lower extremities would be missed, if ABI measurement alone is used for diagnosis of PVD. PMID- 12568207 TI - Effects of angiotensin converting enzyme inhibitor on renal function in patients of membranoproliferative glomerulonephritis with mild to moderate renal insufficiency. AB - AIM: To determine the effect of enalapril, angiotensin-converting-enzyme inhibitor (ACE-I) on progression of renal insufficiency in primary membranoproliferative glomerulonephritis with mild to moderate renal insufficiency. METHOD: Thirty patients with histopathologically proved MPGN having hypertension (grade I and II of JNC-VI criteria of hypertension) and mild to moderate impairment of renal function (creatinine clearance varying from 30-80 ml/min, significant albuminuria and serum creatinine 1.2-3.0 mg/dl) were initially treated with diuretics and 3-blockers to bring down BP < 140/90 mm Hg. These patients were then randomly divided into three groups of 10 each, group I- Control; group II--Nifedipine and group III--Enalapril. In group II and III Nifidepine 30 mg/day and in group III Enalapril 10 mg/day respectively were added in addition and treatment was continued for nine months. These patients were followed up monthly for drug efficacy, side effects and any adverse drug reaction. RESULTS: Out of 30, 28 patients completed the study. At the end of nine months of treatment the patients of control group revealed significant increase in serum creatinine (1.65 +/- 0.38 to 2.17 +/- 0.31 mg/dl), blood urea (34.0 +/- 3.9 to 40.0 +/- 3.1 mg/dl), and 24 hours albuminuria (3.6 +/- 0.6 to 4.2 +/- 0.6 gm) and decrease in creatinine clearance (60.3 +/- 13.3 to 37.5 +/- 11.8 m/min); however, in enalapril group there was decrease in serum creatinine (1.72 +/- 0.45 to 1.24 +/- 0.58 mg/dl), blood urea (34.6 +/- 4.7 to 28.1 +/- 6.7 mg/dl) and 24 hours albuminuria (3.3 +/- 1.0 to 1.6 +/- 1.1 gm) and increase in creatinine clearance (56A +/- 15.8 to 77.1 +/- 23.5 ml/min). The patients on nifedipine showed statistically nonsignificant changes in creatinine clearance, blood urea and serum creatinine; while albuminuria increased from 3.0 +/- 1.3 to 3.9 +/- 0.4 gm/24 hours (p < 0.01). The blood pressure was well controlled in all patients. None of the patient had side effects leading to withdrawal of drugs. No adverse drug reaction was noted. CONCLUSION: ACE-I (enalapril) provided protection against the progression of renal insufficiency in patients of MPGN having hypertension with mild to moderate renal impairment. The renoprotective effects of ACE inhibitor (enalapril) is associated with substantial decrease in albuminuria. PMID- 12568208 TI - Angioplasty for chronic total coronary occlusions: safety and efficacy. AB - OBJECTIVE: To determine the short term results and safety of angioplasty in chronic coronary occlusions. METHODS: Eighty consecutive patients undergoing percutaneous transluminal coronary angioplasty (PTCA) for chronic coronary occlusions were prospectively analyzed for acute success rate and safety of the procedure. RESULTS: The mean age was 46.7 years (range 30-78 years). There were 72 males and eight females. Clinical presentation was recent myocardial infarction (MI) in four cases (5%), unstable angina in 20 (25%), chronic stable angina in 24 (30%) and past history of MI in 32 (40%) cases. Vessel distribution was left anterior descending artery (LAD) in 40 (50%), left circumflex artery (LCx) in 12 (15%) and right coronary artery (RCA) in 28 (35%) cases. Lesion length varied from 8 mm to 37 mm with a mean of 16.7 mm. Acute success rate was 70% (56/80). Twenty four cases (30%) had unsuccessful result due to failure to cross with wire (18 cases) or inability to cross with the balloon (six cases). One major complication in the form of type III coronary perforation was encountered which was successfully managed surgically. CONCLUSION: Percutaneous transluminal coronary angioplasty (PTCA) in chronic total occlusion has a reasonable success rate and very low complication rate. PMID- 12568209 TI - Clinical spectrum of inflammatory myositis in South India--a ten year study. AB - OBJECTIVE: To describe the clinical spectrum of inflammatory myopathies at a referral hospital in South India. METHODS: Patients were assessed for the pattern of muscle involvement, for the presence of arthritis, Raynaud's phenomenon, interstitial lung disease (ILD) and cardiac involvement. Muscle enzymes, electromyogram (EMG) and muscle biopsies were done. RESULTS: Eighty seven patients with inflammatory myopathies were encountered over 10 years. These included 24 with adult polymyositis, 26 with adult dermatomyositis, one with amyopathic dermatomyositis, five with juvenile myositis, one with dermatomysitis following carcinoma breast and 30 with overlap with other connective tissue diseases. There was a female preponderance (M:F = 1:2.35) except in juvenile myosits group (M:F = 1.5:1). The mean age of onset in years was 33.26 in adult polymyositis, 35.03 in adult dermatomyositis, 7.4 in juvenile dermatomyositis, 42 in malignancy-associated dermatomyositis and 25.51 in the overlap group. Proximal muscle weakness was seen in 98.8% patients, dysphagia in 33.3%, distal muscle weakness in 12.5%, respiratory muscle weakness in 9.2% and dysphonia in 4.6%. Other features included arthritis 35.63%, interstitial lung disease (ILD) 9.2%, Raynaud's 5.7%, myocarditis 4.6% and conduction disturbances 1.15%. Eleveated muscle enzymes were seen in 85.1% patients. Eletromyogram was positive in 66.6%. Muscle biopsy was positive in 85.29%. Anti-nuclear antibody was positive in 67.24%. All received steroids, non-responders needed methotrexate (13 patients) or azathioprine (11 patients). Death occurred in 10 (seven with dermatomyositis predominantly due to respiratory involvement and three with overlap). CONCLUSION: There was female preponderance except in juvenile myositis group. Proximal muscle weakness was the commonest feature. ILD was the commonest respiratory problem, while myocarditis was the commonest cardiac problem seen. Response to therapy and prognosis in polymyositis were good with no mortality during the study period. Death in the dermatomyositis group was mainly due to respiratory involvement. PMID- 12568210 TI - Persistent microalbuminuria in type 1 diabetic subjects in South India. AB - AIM: There are only a few reports on the prevalence of microalbuminuria (MAU) in type 1 diabetic subjects in India. This study was done to assess the prevalence of MAU in South Indian diabetic subjects. MATERIAL AND METHODS: MAU was estimated by measuring albumin/creatinine ratio in an early morning urine sample on three occasions using immunoturbidimetry (> 30 microg albumin/mg creatinine) in South Indian patients. Kidney function tests were done in 95 type 1 DM, (M:F, 56:39). Persistent proteinuria was detected in 17 (17.9%) and they were excluded from the study. Further details were taken only in the remaining 78 subjects. Persistent MAU was seen in 22 (28.2%) and other 56 subjects had normoalbuminuria (NAU) (71.8%). RESULTS: There was no significant difference in the age, duration of DM and the average HbAlc between MAU and NAU. MAU developed at the age of > or = 20 years, even in subjects with onset of diabetes at a younger age. Diabetic retinopathy was present in 3 (13.6%) and ECG abnormalities were seen in one patient with MAU. None of the subjects in NAU group had these abnormalities. Prevalence of MAU increased with increasing duration of diabetes, MAU was present in 3 patients (10.3%) with duration of diabetes < or = 5 years, in five patients (23.8%) with duration of diabetes> 5 to < or = 10 years and in 14 (50%) patients with duration of diabetes > 10 years. Hypertension was present in 5 (22.7%) of MAU and only in one subject with NAU. CONCLUSION: It is observed that MAU occurs in type 1 diabetes after the age of 20 years and it is present in a large proportion of the study subjects. PMID- 12568211 TI - The effect of antidepressants on serum melatonin levels in endogenous depression. AB - OBJECTIVES: 1. To assess the serum melatonin levels in patients suffering from endogenous depression and the effect of pharmacological therapy. 2. To establish possible correlation between the height from the mean sea levels and the patients suffering from endogenous depression. METHODS: Forty patients, 18 males and 22 females, suffering from endogenous depression (according to DSM IV criteria), aged 48.3 +/- 12.32 years were evaluated and serum melatonin level was assayed between 9-10 am. They were not on any drugs/medication, which was likely to alter serum melatonin level for one month prior to study. The serum melatonin levels were assessed at 0, 3, 6 and 12 months while they were continued to be treated. It also included 30 controls of the matched age and sex and satisfying the inclusion criteria. The possible correlation was also studied between the serum levels and the height from the mean sea level at which the patients reside. RESULTS: The two groups were comparable at the beginning of the study. The serum melatonin levels started falling from three months onwards (from 9.99 +/- 3.59 pg/nl to 8.49 +/- 3.16 pg/nl; p < 0.05). However, the decline was maximum between 3-6 months (from 8.49 +/- 3.16 to 5.589 +/- 1.96; p<0.05). The serum levels became stationary beyond six months. Highest melatonin levels were observed in patients residing at an altitude of 6001-8000 metres (14.32 +/- 2.68 pg/ml; p < 0.05) followed by 4,001-6,000 meters (11.137 +/- 2.62 pg/ml; p < 0.05). However, the levels were almost stationary below 4,000 metres (p = 0.05). CONCLUSION: 1. Higher serum melafonin values were observed in patients suffering from endogenous depression. 2. Morning serum melatonin values decreased with pharmacological therapies. 3. Patients living at higher altitudes had higher serum values for the hormone. PMID- 12568212 TI - Evaluation of communication skills of physicians about asthma. AB - OBJECTIVE: Communication skills in medical practice are important. To assess the communication skills of physicians about asthma, we undertook this study. METHODS: One thousand four hundred and twenty one physicians attending the annual conference of association of physicians of India volunteered to participate in the study. They were divided into three categories. 'Teachers' group 1, (n = 131), 'Clinicians', group 2, (n = 951) and 'Students' group 3, (n = 339). Questionnaire containing ten most frequently asked questions by asthmatic patients were given to the physicians. A panel of three patients assessed the replies of these question and graded them as 'Convincing', 'Just convincing' and 'Not convincing'. RESULTS: Replies of only 2% of physicians were graded as convincing, 15.6% just convincing and 82.4 as not convincing. Among group 1 none gave a convincing reply while replies of 2.2% of group 2 and 2.4% of group 3 were graded as convincing. The difference in results was not significant statistically on comparing the three groups (p > 0.1). CONCLUSIONS: Communication skills are the integral part of patient care and management. Our study demonstrated lack of effective communication skills among physicians. PMID- 12568213 TI - Angiotensin and its inhibition: a nephrologist point of view. PMID- 12568214 TI - Sir Ronald Ross(1857-1932)--first day cover & postal-stamp released in India- 1997. PMID- 12568215 TI - Dermatomyositis. PMID- 12568216 TI - Diffuse parenchymal lung disease: an approach to diagnosis. PMID- 12568217 TI - Current recommendations on iron therapy in anaemia of chronic renal failure. PMID- 12568218 TI - Cellular and molecular biology of adiposity and adipogenesis. PMID- 12568219 TI - Baclofen in the treatment of intractable hiccups. AB - Availability of a large number of drugs/remedies for the management of hiccups suggests that all the available drugs are not effective in all the patients. Baclofen was successful in relieving the intractable hiccups where all other known drugs of hiccup were unable to provide relief. PMID- 12568220 TI - Herpes simplex encephalitis in acquired immune deficiency syndrome. PMID- 12568221 TI - Indirect hyperbilirubinemia with indinavir. AB - Indinavir is a protease inhibitor used in the treatment of HIV infected individuals and as post-exposure prophylaxis. Indinavir is associated with various adverse effects including gastrointestinal, a lipodystrophy syndrome and nephrolithiasis. We describe indirect hyperbilirubinemia as an adverse effect of indinavir in a person on post-exposure prophylaxis (PEP). PMID- 12568222 TI - Painless pancreatitis: a rare manifestation of leptospirosis. AB - A rare case of painless pancreatitis caused by Leptospira is reported. The patient presented clinically with features of paralytic ileus and renal failure. Hyperamylasemia was persistent inspite of recovery of renal function. The patient recovered on treatment with penicillin and serum amylase returned to normal in three months. PMID- 12568223 TI - Lymphocytic interstitial lung disorder: an isolated entity. AB - A 40 years female patient presented with recurrent haempoptysis since last five years and taking antituberculous as well as antidiabetic treatment. She was further investigated and found having lymphocytic interstitial lung disease without any other autoimmune disorder. She was treated by surgery with good response. PMID- 12568224 TI - Acute febrile neutrophilic dermatosis following tuberculous infection. AB - A case of acute febrile neutrophilic dermatosis (Sweet's syndrome) in association with tuberculosis has been reported. However association of the two diseases is not common. It appears as if Mycobacterium tuberculosis present somewhere in the body can induce cutaneous reactions as is the case in erythema nodosum and erythema induratum. PMID- 12568225 TI - Unilateral pulmonary agenesis with ipsilateral musculoskeletal anomalies. AB - A young patient with right pulmonary agenesis is described. She presented with left pneumonitis and improved with antibiotic therapy. She additionally had absence of right thumb and reduced muscle mass over right side of face. The anomalies may have resulted from an antenatal insult around the fourth week of embryonic development. PMID- 12568226 TI - Hypokalemic paralysis in pregnancy. AB - Hypokalemic paralysis is an uncommon entity with varied etiology. Renal tubular acidosis is an unusual cause of this disorder. Its occurrence with pregnancy is rare. We report such a case and mention the implications of the association and review relevant literature breifly. PMID- 12568227 TI - Poisonous jellyfish contact and paralytic ileus. PMID- 12568228 TI - Non-secretory multiple myeloma with hypercalcemic acute renal failure. PMID- 12568229 TI - Spinal tuberculosis. PMID- 12568230 TI - Idiopathic CD4+ T-lymphocytopenia. PMID- 12568231 TI - Diagnostic dilemma: aspergillosis. PMID- 12568232 TI - Cyclosporine-induced reversible pure motor spastic paraparesis in a patient with aplastic anaemia. PMID- 12568233 TI - Unusual distant metastases from papillary thyroid carcinoma. PMID- 12568234 TI - Diabetes mellitus and G-protein dysfunction: facts and possibilities. PMID- 12568235 TI - Intercostal lung herniation in cavitary pulmonary tuberculosis. PMID- 12568236 TI - Oral contraceptives and central retinal vein occlusion. PMID- 12568237 TI - Drug treatment of mild hypertension in elderly--we should be cautious in some patients. PMID- 12568238 TI - Efficacy and safety of monoclonal anti-D immunoglobulin in comparison with polyclonal anti-D immunoglobulin in prevention of rho isoimmunization. PMID- 12568239 TI - Stands to reason. PMID- 12568240 TI - Awakening to the behavioral analysis of sleep in Drosophila. AB - Perhaps the most observable of the many circadian oscillations that have been described in both vertebrate and invertebrate animals is the daily alterations in periods of rest and activity. Recent studies in the fruit fly Drosophila melanogaster suggest that these periods of inactivity are not simply rest but share many of the fundamental components that define mammalian sleep. Thus, quiescent episodes are characterized by reduced awareness of the environment and are homeostatically regulated. Although this field is in its infancy, recent studies have focused on the interaction between circadian and homeostatic processes. These results indicate that components of the circadian clock may play a substantial role in mechanisms underlying sleep homeostasis at the molecular level. In this article, the author reviews recent advances obtained using Drosophila as a model system to elucidate fundamental components of sleep regulation. PMID- 12568241 TI - Gender dimorphism in the role of cycle (BMAL1) in rest, rest regulation, and longevity in Drosophila melanogaster. AB - The central clock is generally thought to provide timing information for rest/activity but not to otherwise participate in regulation of these states. To test the hypothesis that genes that are components of the molecular clock also regulate rest, the authors quantified the duration and intensity of consolidated rest and activity for the four viable Drosophila mutations of the central clock that lead to arrhythmic locomotor behavior and for the pdf mutant that lacks pigment-dispersing factor, an output neuropeptide. Only the cycle (cyc01) and Clock (Clk(Jrk)) mutants had abnormalities that mapped to the mutant locus, namely, decreased consolidated rest and grossly extended periods of activity. All mutants with the exception of the cyc01 fly exhibited a qualitatively normal compensatory rebound after rest deprivation. This abnormal response in cyc01 was sexually dimorphic, being reduced or absent in males and exaggerated in females. Finally, the cyc01 mutation shortened the life span of male flies. These data indicate that cycle regulates rest and life span in male Drosophila. PMID- 12568242 TI - Defined cell groups in the rat suprachiasmatic nucleus have different day/night rhythms of single-unit activity in vivo. AB - The electrical activity of the rat suprachiasmatic nucleus (SCN) was examined in anesthetized rats in vivo using single-unit electrophysiological techniques. The present data confirm the daily variation in the electrical activity of the SCN previously reported in vitro and in vivo using multiple-unit recording techniques. They further suggest that subpopulations of suprachiasmatic neurons with different neural connections have a different daily rhythm of activity. Neurons in the SCN region showed a significant rhythm of activity (p = 0.034; Kruskall-Wallis analysis of variance [KW-ANOVA]). The greatest activity occurred during the second part of the light period (ZT 10-12), and the lowest activity occurred in the early part of the light period (ZT 0-2). The subgroup of cells in the suprachiasmatic region with output projections to the arcuate nucleus (ARC) and/or supraoptic nucleus (SON) regions also showed a significant rhythm (p = 0.001; K-W ANOVA). Their activity appeared to show two peaks near the light-dark (ZT 10-12) and dark-light (ZT 22-24) transition periods with the lowest activity at ZT 16-18. This rhythm was significantly different (p = 0.016) from that of neurons without an output projection to the ARC and/or SON. Retinorecipient suprachiasmatic neurons appeared to have a less robust daily rhythm in their activity. The change in the firing behavior of the cells was not reflected simply by changes in mean firing rate. Examination of the coefficient of variation of the interspike interval distribution of cells at different times of day revealed changes in the firing pattern of cells in the SCN region that did not have output projections (p = 0.032; K-W ANOVA). The present results thus suggest that the SCN is composed of a heterogeneous population of neurons and that different rhythms of activity are expressed by neurons with different neural connections. There were changes in both firing pattern and firing rate. PMID- 12568243 TI - Testosterone suppresses circadian responsiveness to social cues in the diurnal rodent Octodon degus. AB - The diurnal, social rodent Octodon degus displays a robust sex difference in the ability to use social cues to facilitate reentrainment following a phase advance of the light cycle. Adult females housed with a female social cue donor reentrained 25% to 40% faster than did females reentraining alone. However, reentrainment rates of males were unaffected by exposure to female social cues during reentrainment. The authors hypothesized that males were less sensitive to the reentrainment-enhancing effects of social cues and that their higher threshold to the stimuli could be overcome if the social cues were either increased in strength or salience. Housing a male with two females significantly shortened the time to reentrain following a 9-h phase advance (p = 0.002). Housing with a sister had no effect on reentrainment. Therefore, male degus are able to respond to social cues but require the stimulus to be stronger than that for females. The effect of testosterone was tested by comparing reentrainment rates of castrated males before and after testosterone replacement both with and without a female social cue donor. Castrated males responded to a single female social cue donor, reentraining 35% faster than when housed alone (p = 0.006), whereas the time to reentrainment of intact males and males with testosterone capsule implants did not differ. Intact females were also implanted with testosterone and phase shifted with and without donors. Testosterone treatment eliminated the increase in reentrainment rates in the presence of social cues. The authors conclude that the rate of recovery from odor-enhanced phase shifts is modulated by activational effects of testosterone in male degus. Testosterone is also effective in suppressing social cue responsiveness in females, suggesting that testosterone's effects on responsiveness are not sexually dimorphic. This hormonal effect likely occurs by altering sensory system functions or CNS response to sensory information. PMID- 12568245 TI - Photoperiod affects amplitude but not duration of in vitro melatonin production in the ruin lizard (Podarcis sicula). AB - The pineal gland and its major output signal melatonin have been demonstrated to play a central role in the seasonal organization of the ruin lizard Podarcis sicula. Seasonal variations in the amplitude of the nocturnal melatonin signal, with high values in spring as compared to low values in summer and autumn, have been found in vivo. The authors examined whether the pineal gland of the ruin lizard contains autonomous circadian oscillators controlling melatonin synthesis and whether previously described seasonal variations of in vivo melatonin production can also be found in isolated cultured pineal glands obtained from ruin lizards in summer and winter. In vitro melatonin release from isolated pineal glands of the ruin lizard persisted for 4 days in constant conditions. Cultured explanted pineal glands obtained from animals in winter and summer showed similar circadian rhythms of melatonin release, characterized by damping of the amplitude of the melatonin rhythm. Although different photoperiodic conditions were imposed on ruin lizards before explantation of pineal glands, the authors did not find any indication for corresponding differences in the duration of elevated melatonin in vitro. Differences were found in the amplitude of in vitro melatonin production in light/dark conditions and, to a lesser degree, in constant conditions. The presence of a circadian melatonin rhythm in vitro in winter, although such a rhythm is absent in vivo in winter, suggests that pineal melatonin production is influenced by an extrapineal oscillator in the intact animal that may either positively or negatively modulate melatonin production in summer and winter, respectively. PMID- 12568244 TI - Photoperiodic adjustments in immune function protect Siberian hamsters from lethal endotoxemia. AB - Seasonal changes in day length enhance or suppress components of immune function in individuals of several mammalian species. Siberian hamsters (Phodopus sungorus) exhibit multiple changes in neuroendocrine, reproductive, and immune function after exposure to short days. The manner in which these changes are integrated into the host response to pathogens is not well understood. The present experiments tested the hypothesis that short-day changes in immune function alter the pathogenesis of septic shock and survival after challenge with endotoxin. Male and female Siberian hamsters raised in long-day photoperiods were transferred as adults to short days or remained in their natal photoperiod. Six to 8 weeks later, hamsters were injected i.p. with 0, 1, 2.5, 10, 25, or 50 mg/kg bacterial lipopolysaccharide (LPS) (the biologically active constituent of endotoxin), and survival was monitored for 96 h. Short days significantly improved survival of male hamsters treated with 10 or 25 mg/kg LPS and improved survival in females treated with 50 mg/kg LPS. Transfer from long to short days shifted the LD50 in males by approximately 90%, from 5.3 to 9.9 mg/kg, and in females from 11.1 to 15.0 mg/kg (+35%). Long-day females were more resistant than were males to lethal endotoxemia. In vitro production of the proinflammatory cytokine TNFalpha in response to LPS stimulation was significantly lower in macrophages extracted from short-day relative to long-day hamsters, as were circulating concentrations of TNFalpha in vivo after i.p. administration of LPS, suggesting that diminished cytokine responses to LPS in short days may mitigate the lethality of endotoxemia. Adaptation to short days induces changes in immune parameters that affect survival in the face of immune challenges. PMID- 12568247 TI - Life between clocks: daily temporal patterns of human chronotypes. AB - Human behavior shows large interindividual variation in temporal organization. Extreme "larks" wake up when extreme "owls" fall asleep. These chronotypes are attributed to differences in the circadian clock, and in animals, the genetic basis of similar phenotypic differences is well established. To better understand the genetic basis of temporal organization in humans, the authors developed a questionnaire to document individual sleep times, self-reported light exposure, and self-assessed chronotype, considering work and free days separately. This report summarizes the results of 500 questionnaires completed in a pilot study individual sleep times show large differences between work and free days, except for extreme early types. During the workweek, late chronotypes accumulate considerable sleep debt, for which they compensate on free days by lengthening their sleep by several hours. For all chronotypes, the amount of time spent outdoors in broad daylight significantly affects the timing of sleep: Increased self-reported light exposure advances sleep. The timing of self-selected sleep is multifactorial, including genetic disposition, sleep debt accumulated on workdays, and light exposure. Thus, accurate assessment of genetic chronotypes has to incorporate all of these parameters. The dependence of human chronotype on light, that is, on the amplitude of the light:dark signal, follows the known characteristics of circadian systems in all other experimental organisms. Our results predict that the timing of sleep has changed during industrialization and that a majority of humans are sleep deprived during the workweek. The implications are far ranging concerning learning, memory, vigilance, performance, and quality of life. PMID- 12568246 TI - Inferior retinal light exposure is more effective than superior retinal exposure in suppressing melatonin in humans. AB - Illumination of different areas of the human retina elicits differences in acute light-induced suppression of melatonin. The aim of this study was to compare changes in plasma melatonin levels when light exposures of equal illuminance and equal photon dose were administered to superior, inferior, and full retinal fields. Nine healthy subjects participated in the study. Plexiglass eye shields were modified to permit selective exposure of the superior and inferior halves of the retinas of each subject. The Humphrey Visual Field Analyzer was used both to confirm intact full visual fields and to quantify exposure of upper and lower visual fields. On study nights, eyes were dilated, and subjects were exposed to patternless white light for 90 min between 0200 and 0330 under five conditions: (1) full retinal exposure at 200 lux, (2) full retinal exposure at 100 lux, (3) inferior retinal exposure at 200 lux, (4) superior retinal exposure at 200 lux, and (5) a dark-exposed control. Plasma melatonin levels were determined by radioimmunoassay. ANOVA demonstrated a significant effect of exposure condition (F = 5.91, p < 0.005). Post hoc Fisher PLSD tests showed significant (p < 0.05) melatonin suppression of both full retinal exposures as well as the inferior retinal exposure; however, superior retinal exposure was significantly less effective in suppressing melatonin. Furthermore, suppression with superior retinal exposure was not significantly different from that of the dark control condition. The results indicate that the inferior retina contributes more to the light-induced suppression of melatonin than the superior retina at the photon dosages tested in this study. Findings suggest a greater sensitivity or denser distribution of photoreceptors in the inferior retina are involved in light detection for the retinohypothalamic tract of humans. PMID- 12568248 TI - Method for predicting photocatalytic oxidation rates of organic compounds. AB - In designing a photocatalytic oxidation (PCO) system for a given air pollution source, destruction rates for volatile organic compounds (VOCs) are required. The objective of this research was to develop a systematic method of predicting PCO rate constants by correlating rate constants with physical-chemical characteristics of compounds. Accordingly, reaction rate constants were determined for destruction of volatile organics over a titanium dioxide (TiO2) catalyst in a continuous mixed-batch reactor. It was found that PCO rate constants for alkanes and alkenes vary linearly with gas-phase ionization potential (IP) and with gas-phase hydroxyl radical reaction rate constant. The correlations allow rates of destruction of compounds not tested in this research to be predicted based on physical-chemical characteristics. PMID- 12568250 TI - Pig slurry concentration by vacuum evaporation: influence of previous mesophilic anaerobic digestion process. AB - Water can be removed from pig slurry by evaporation, through the application of wasted heat from a power plant or from other processes. Apart from obtaining a concentrate with an obviously higher nutrient concentration than the original slurry, another objective of water removal is to obtain water as condensate, which could be reused. The objective of this work was to study the vacuum evaporation of pig slurry liquid fraction and to evaluate condensate composition as a function of both pH (4, 5, and 6) and pig slurry type (fresh slurry and anaerobically digested slurry). Batch experiments showed that condensate characteristics, total ammonia nitrogen (NH3-N), volatile fatty acids (VFA), and chemical oxygen demand were strongly dependent on initial slurry pH. In addition to producing part of the required thermal energy, previous anaerobic digestion presented several other clear advantages. The consumption of VFA and other volatile organic compounds during anaerobic digestion reduced the volatilization of organic matter in the evaporation treatment and, consequently, provided a higher quality condensate. PMID- 12568249 TI - Gaseous and particulate emissions from a DC arc melter. AB - Tests treating soils contaminated with metal compounds and radionuclide surrogates were conducted in a DC arc melter. The soil melted, and glassy or ceramic waste forms with a separate metal phase were produced. Tests were run in the melter plenum with either air or N2 purge gases. In addition to nitrogen, the primary emissions of gases were CO2, CO, oxygen, methane, and oxides of nitrogen (NO(x)). Although the gas flow through the melter was low, the particulate concentrations ranged from 32 to 145 g/m3. Cerium, a nonradioactive surrogate for plutonium and uranium, was not enriched in the particulate matter (PM). The PM was enriched in cesium and highly enriched in lead. PMID- 12568251 TI - Compatibility analysis of material and energy recovery in a regional solid waste management system. AB - The rising prices of raw materials and concerns about energy conservation have resulted in an increasing interest in the simultaneous recovery of materials and energy from waste streams. Compatibility exists for several economic, environmental, and managerial reasons. Installing an on-site or off-site presorting facility before an incinerator could be a feasible alternative to achieve both goals if household recycling programs cannot succeed in local communities. However, the regional impacts of presorting solid waste on a waste to-energy facility remain unclear because of the inherent complexity of solid waste compositions and properties over different areas. This paper applies a system-based approach to assess the impact of installing a refuse-derived fuel (RDF) process before an incinerator. Such an RDF process, consisting of standard unit operations of shredding, magnetic separation, trommel screening, and air classification, might be useful for integrating the recycling and presorting efforts for a large-scale municipal incinerator from a regional sense. An optimization modeling analysis is performed to characterize such integration potential so that the optimal size of the RDF process and associated shipping patterns for flow control can be foreseen. It aims at exploring how the waste inflows with different rates of generation, physical and chemical compositions, and heating values collected from differing administrative districts can be processed by either a centralized presorting facility or an incinerator to meet both the energy recovery and throughput requirements. A case study conducted in Taipei County, which is one of the most densely populated metropolitan areas in Taiwan, further confirms the application potential of such a cost-benefit analysis. PMID- 12568252 TI - Interpretation of air pollution mortality: number of deaths or years of life lost? AB - This paper examines the relation between the results of epidemiologic studies of air pollution mortality and impact indicators that can be informative for environmental policy decisions. Using models that are simple and transparent, yet contain the essential features, it is shown that (1) number of deaths is not meaningful for air pollution, whereas loss of life expectancy (LLE) is an appropriate impact indicator; (2) the usual short-term (time series) studies yield a change in daily number of deaths attributable to acute effects of pollution, without any information on the associated LLE (although some information on this has recently become available by extending the observation window of time series); and (3) long-term studies yield a change in age-specific mortality, which makes it possible to calculate the total population averaged LLE (acute and chronic effects) but not the total number of premature deaths attributable to air pollution. The latter is unobservable because one cannot distinguish whether few individuals suffer a large or many suffer a small LLE. The paper calculates the LLE from exposure to PM10, as implied by the long-term mortality studies of adults and infants; population LLE for infants turns out to be an order of magnitude smaller than for adults. The LLE implied by short-term studies is a small fraction of the total loss implied by long-term studies, even if one assumes a very high loss per death. Applied to environmental policy, taking a permanent 50-70% reduction of PM10 as a reasonable goal, one finds a corresponding increase of average life expectancy in urban areas of the European Union (EU) and the United States of approximately four months. PMID- 12568253 TI - Odor concentration decay and stability in gas sampling bags. AB - This paper presents results of an experimental study into factors contributing to decay of odor samples during storage, between 4 and 40 hr after sample collection. The odor studied was sampled from a tobacco processing plant as part of collaborative research with a view to establishing a manual outlining methods for odor annoyance management, specifically for the tobacco industry. In August and September 1997, an experimental program was carried out in which two types of tobacco odor were sampled: Burley Toaster and Mix. The dependent variable was odor concentration in the bag, measured by dynamic olfactometry in accordance with the draft Comite Europeen de Normalisation (CEN) standard EN13725 "Air Quality-Determination of Odor Concentration by Dynamic Olfactometry." The independent variables were sampling bag material, degree of dilution during sampling, dilution gas used, particle removal during sampling, and age of sample in hours. In the first phase, 94 odor analyses were carried out. In a second test, 32 samples were analyzed for odor concentration. In addition, 16 samples were analyzed using gas chromatography-mass spectrometry (GC-MS). Analysis of the results (analysis of variance) led to the unexpected conclusion that Nalophan film bags performed significantly better than metalized Cali-Bond layered film as a bag material. The odor concentration of samples in Nalophan bags remained relatively stable between 4 and 12 hr after sampling. After 30 hr, decay to about half the initial concentration, as measured at 4 hr, was observed. Particle removal during sampling caused the odor concentration in the bags to be reduced by approximately 20%. For practical reasons, particle removal remains useful, to avoid contamination of equipment. Using air or nitrogen as the neutral gas for pre-dilution during sampling or the dilution factor used (between factor 2 and 6) did not appear to have an effect on the decay characteristic of odor samples. The following recommendations are suggested for the practice of collecting odor samples and apply specifically to tobacco processing emissions: Analyze samples as soon as possible, preferably within 12 hr; When samples age for more than 12 hr, decay is likely to cause a reduction in odor concentration to half the original concentration at age 30 hr; Use sampling bags made of Nalophan NA or benchmark performance of other materials against Nalophan NA before using alternative materials; Use pre-dilution when sampling only for the purpose of avoiding condensation during sample storage. Use an appropriate minimum dilution factor to avoid condensation; Both nitrogen and high-purity (synthetic) air are suitable to use as neutral gas for pre-dilution; and When sampling tobacco odors, use an odorless filter to remove particles. This practice removes a source of variation and avoids contamination of equipment. The effect on results, despite being consistently lower in odor concentration, is not meaningful in terms of perceived intensity or annoyance potential. PMID- 12568254 TI - Fate and potential environmental effects of methylenediphenyl diisocyanate and toluene diisocyanate released into the atmosphere. AB - Information from a variety of sources has been collected and summarized to facilitate an overview of the atmospheric fate and potential environmental effects of emissions of methylenediphenyl diisocyanate (MDI) or toluene diisocyanate (TDI) to the atmosphere. Atmospheric emissions of both MDI and TDI are low, both in terms of concentration and mass, because of their low volatility and the need for careful control over all aspects of their lifecycle from manufacture through disposal. Typical emission losses for TDI are 25 g/t of TDI used in slabstock foam production. MDI emission losses are lower, often less than 1 g/t of MDI used. Dispersion modeling predicts that concentrations at the fenceline or beyond are very low for typical releases. Laboratory studies show that TDI (and by analogy MDI) does not react with water in the gas phase at a significant rate. The primary degradation reaction of these aromatic diisocyanates in the atmosphere is expected to be oxidation by OH radicals with an estimated half-life of one day. Laboratory studies also show that this reaction is not expected to result in increased ground-level ozone accumulation. PMID- 12568255 TI - Comparison of vehicle exhaust emissions from modified diesel fuels. AB - Three diesel fuels, one oil sand-derived (OSD) diesel serving as base fuel, one cetane-enhanced base fuel, and one oxygenate [diethylene glycol dimethyl ether (DEDM)]-blended base fuel, were tested for their emission characterizations in vehicle exhaust on a light-duty diesel truck that reflects the engine technology of the 1994 North American standard. Both the cetane-enhanced and the oxygenate blended fuels were able to reduce regulated [CO, particulate matter (PM), total hydrocarbon (THC)] and nonregulated [polyaromatic hydrocarbons (PAHs), carbonyls, and other volatile organic chemicals] emissions, except for nitrogen oxides (NO(x)), compared with the base fuel. Although burning a fuel that contains oxygen could conceivably yield more oxygenated compounds in emissions, the oxygenate-blended diesel fuel resulted in reduced emissions of formaldehyde along with hydrocarbons such as benzene, 1,3-butadiene, and PAHs. Reductions in nitro PAH emissions have been observed in both the cetane-enhanced and oxygenated fuels. This further demonstrates the benefits of using a cetane enhancer and the oxygenated fuel component. PMID- 12568256 TI - Utilization of coal fly ash as a slow-release granular medium for soil improvement. AB - This work proposes a new potential application of waste coal fly ash as a K fertilizer support. Fly ash was reacted with KOH to facilitate the impregnation of K as well as to enhance the bonding force. In particular, the applied process resulted in a significant slow-releasing characteristic of fertilizer elements. To examine the effect of K impregnation, a few detailed leaching tests were carried out in terms of process variables such as reaction time and temperature, sintering time and temperature, and KOH concentration. The current experiment presented an optimum preparation condition that is competitive with conventional commercial fertilizers. The manufactured ash fertilizers inhibited release of the K elements. It was also found through the continuous leaching test with pure water that the ash fertilizer had excellent moisture absorbability. However, the effects of some trace elements in fly ash on soil health and crop productivity as well as environmental considerations need to be established with long-term studies. PMID- 12568257 TI - Short-term temporal variation in PM2.5 mass and chemical composition during the Atlanta Supersite Experiment, 1999. AB - Measurements in urban Atlanta of transient aerosol events in which PM2.5 mass concentrations rapidly rise and fall over a period of 3-6 hr are reported. The data are based on new measurement techniques demonstrated at the U.S. Environmental Protection Agency (EPA) Atlanta Supersite Experiment in August 1999. These independent instruments for aerosol chemical speciation of NO3-, SO4(2-), NH4+, and organic and elemental carbon (OC and EC), reconstructed the observed hourly dry PM2.5 mass to within 20% or better. Data from the experiment indicated that transient PM2.5 events were ubiquitous in Atlanta and were typically characterized by a sudden increase of EC (soot) and OC in the early morning or SO4(2-) in the late afternoon. The frequent temporal decoupling of these events provides insights into their origins, suggesting mobile sources in metro Atlanta as the main contributor to early morning PM2.5 and more regionally located point SO2 sources for afternoon PM2.5 events. The transient events may also have health implications. New data suggest that short-term PM2.5 exposures may lead to adverse health effects. Standard integrated filter-based techniques used in PM2.5 compliance monitoring networks and in most past PM2.5 epidemiologic studies collect samples over 24-hr periods and thus are unable to capture these transient events. Moreover, health-effects studies that focus on daily PM2.5 mass alone cannot evaluate the health implications of the unique and variable chemical properties of these episodes. PMID- 12568258 TI - Removal of ammonia by biofilters: a study with flow-modified system and kinetics. AB - The characteristics of ammonia removal by two types of biofilter (a standard biofilter with vertical gas flow and a modified biofilter with horizontal gas flow) were investigated. A mixture of organic materials such as compost, bark, and peat was used as the biofilter media based on the small-scale column test for media selection. Complete removal capacity, defined as the maximum inlet load of ammonia that was completely removed, was obtained. The modified biofilter showed complete removal up to 1.0 g N/kg dry material/day. However, the removal capacity of the standard biofilter started to deviate from complete removal around 0.4 g N/kg dry material/day, indicating that the modified biofilter system has higher removal efficiency than the standard upflow one. In kinetic analysis of the biological removal of ammonia in each biofilter system, the maximum removal rate, Vm, was 0.93 g N/kg dry material/day and the saturation constant, Ks, was 32.55 ppm in the standard biofilter. On the other hand, the values of Vm and Ks were 1.66 g N/kg dry material/day and 74.25 ppm, respectively, in the modified biofilter system. PMID- 12568259 TI - Source profiles and ozone formation potentials of volatile organic compounds in three traffic tunnels in Kaohsiung, Taiwan. AB - Twenty-five volatile organic compounds (VOCs) up to C10 were measured using Carbotrap multibed thermal adsorption tubes during the morning and afternoon rush hours on four different days in all three traffic tunnels in Kaohsiung, Taiwan. A gas chromatograph (GC) equipped with a flame-ionization detector (FID) was then used to analyze the VOCs. The analytical results show that VOC concentrations increase with traffic flow rate, and emission profiles in the three tunnels are mostly in the range C2-C6. In addition to the traffic conditions and vehicle type, the pattern of emissions in each tunnel was also influenced by other factors, such as vehicle age, nearby pollution sources, and the spatial or temporal variation of VOCs in the urban atmosphere. The ozone formation potential (OFP) in each tunnel was assessed based on the maximum incremental reactivities of the organic species, demonstrating that OFP increases with traffic flow rate. Vehicle distribution influences the contributions of organic group to OFP in a tunnel. Meanwhile, when ranked in descending order of contribution to OFP in all tunnels, the organic groups followed the sequence olefins, aromatics, and paraffins. PMID- 12568260 TI - The short-term outcome of surgical treatment for painful varus arthritis in association with chronic ACL deficiency. AB - A review of patients with anterior cruciate ligament (ACL) insufficiency, symptomatic medial compartment osteoarthritis, and varus malalignment of the knee was performed. Twenty-six patients met the inclusion criteria. Twelve patients were treated with a valgus closing-wedge high tibial osteotomy (group 1). Fourteen patients were treated with a valgus closing-wedge high tibial osteotomy combined with arthroscopic ACL reconstruction (group 2). Twenty-five patients were available for follow-up at a minimum of 2 years. For group 1 patients, high tibial osteotomy alone had no effect on the Lachman test or pivot shift phenomena. For group 2 patients, combined high tibial osteotomy/ACL reconstruction resulted in a grade 1 Lachman test in 11 of 13 patients, and a negative pivot shift in 12 of 13 patients. No deficits in range of motion were noted in either group. Prior to surgery, 14 (56%) patients participated in recreational sports; 23 (92%) patients were able to participate in recreational sports at follow-up. Radiographs demonstrated osteoarthritic progression in group 1 and 2 patients (P<.05). The results of this study suggest that high tibial osteotomy alone and combined high tibial osteotomy/ACL reconstructions are effective in the surgical treatment of varus, ACL-deficient knees with symptomatic medial compartment arthritis; however, good or excellent results were more often seen after the combined procedure. PMID- 12568261 TI - Osteochondral autograft plug transfer: achieving perpendicularity. AB - Although osteochondral autograft plug transfer is a common clinical procedure, few authors have addressed the outcomes or how variations and surgical technique can influence results. Variations in arthroscopic versus open harvesting techniques, as well as intercondylar notch versus lateral trochlear ridge harvest locations, influence the ability to obtain the most perpendicular osteochondral grafts. Eight fresh-frozen cadaveric knees were studied. Four plugs were harvested from each knee, two from the superior lateral intercondylar notch area and two from the superior lateral trochlear ridge area. Plugs harvested from the lateral trochlear ridge (86.9 degrees) were statistically more perpendicular than the plugs harvested from the intercondylar notch (83.3 degrees) (P=.02). No statistically significant difference was noted between the open (86.4 degrees) and arthroscopic (83.8 degrees) harvesting techniques (P=.11). PMID- 12568262 TI - High tibial valgus osteotomy for medial gonarthrosis: a 10- to 21-year study. AB - This retrospective study reviewed the long-term experience with high tibial osteotomy and determined which factors influence the results. Between 1980 and 1989, 120 closing wedge high tibial osteotomies for varus gonarthrosis were performed in 102 patients. Twenty-nine knees were excluded because the patients died (17 knees), were bedridden (7 knees), or lost to follow-up (5 knees). Thirty of the remaining 91 knees had a conversion to total knee replacement (TKR) after 11 years on average, leaving 61 knees with a high tibial osteotomy available for clinical and radiographic evaluation at an average follow-up of 15 years (range: 10-21 years). Of the 91 knees, excellent/good results were found in 49% and fair/poor in 51%. Anatomical femorotibial angle in the 61 knees at follow-up averaged 4.7 degrees +/- 5 degrees of valgus (range: 3 degrees varus to 23 degrees valgus). Alignment obtained at consolidation changed with varus recurrence at follow-up in 14% of 61 knees and did not correlate with the clinical results. Twelve (19%) knees showed a patella baja (Caton ratio <0.6) at follow-up, which correlated with patients immobilized postoperatively by a cylinder cast (P=.04). A valgus alignment at consolidation between 8 degrees and 15 degrees, good muscle strength, and male gender correlated with better results (P<.05). Survivorship analysis, considering an unsatisfactory result or revision to TKR as the endpoint, was 96% at 5 years, 88% at 7 years, 78% at 10 years, and 57% at 15 years. High tibial osteotomy provides symptomatic relief for approximately 10 years, but is unlikely to provide permanent relief. PMID- 12568263 TI - Comparison between the shape of resected femoral sections and femoral prostheses used in total knee arthroplasty in Japanese patients: simulation using three dimensional computed tomography. AB - Simulation of the femoral cut for total knee arthroplasty was performed in 44 knees using three-dimensional computed tomography. The three-dimensional images were measured, and the shape of the femur was compared to the provided femoral prosthesis. The ratio between the medial-lateral and anteroposterior dimensions of the three-dimensional images did not always match the prosthesis. The widths of the medial condyles of the three-dimensional images tended to be larger than those of the prosthesis. The lengths of the lateral posterior condyles of the three-dimensional images tended to be shorter than those of the prosthesis. These results suggest that a new prosthesis should be designed to achieve a better anatomical fit. PMID- 12568264 TI - Correction of large bilateral tibia vara with the Ilizarov method. PMID- 12568265 TI - Double-plating nonunions of high tibial osteotomies. PMID- 12568266 TI - The medial parapatellar approach to the knee. PMID- 12568267 TI - The midvastus approach to the knee. PMID- 12568268 TI - The subvastus approach to the knee. PMID- 12568269 TI - The quadriceps snip for exposing the stiff knee. PMID- 12568270 TI - Tibial tubercle osteotomy. AB - During primary and revision TKA, difficulties with exposure may be due to poor motion, obesity, and patellar baja. To gain accurate component positioning and avoid catastrophic complications with the extensor mechanism, a stepwise approach to optimizing the exposure should be used. In cases where the standard medial parapatellar arthrotomy is inadequate, a quadriceps snip is helpful. Occasionally, exposure will still be limited and in these cases, a tibial tubercle osteotomy can be used following the quadriceps snip. The tubercle osteotomy consistently heals postoperatively and results in less extensor lag than the V-Y quadricepsplasty. To minimize postoperative complications, strict attention to the operative technique should be maintained. PMID- 12568271 TI - The lateral approach for total knee arthroplasty. PMID- 12568272 TI - Effectiveness of auricular therapy on sleep promotion in the elderly. AB - Sleep disturbances are a particularly common problem in the elderly. The purpose of this study was to examine the effectiveness of auricular therapy on sleep behaviors in the elderly. One hundred and twenty participants of 60 years old or above and who were suffering from sleep disturbances were invited to participate in this study. Eligible participants were randomly allocated to receive auricular therapy using Junci Medulla (Group A = 30), Semen Vaccariae (Group B = 30) or magnetic pearls (Group C = 60). Groups A and B were the control groups, while Group C was the experimental group. Seven auricular points which are thought to have an effect on promoting sleep were selected. The total treatment course lasted for three weeks. Objective measurement using actigraphic monitoring was performed before the therapy commenced, in the middle period of the therapy, and within one week after the therapy had been completed. After the therapy, there were significant differences among the three groups in terms of the nocturnal sleep time (NST) (F(2,117) = 6.84, p < 0.05) and sleep efficiency (SE) (F(2,117) = 7.69, p < 0.05). Significant improvement in the sleep behaviors was observed in the experimental group using magnetic pearls. In a backward multiple regression, the effect of auricular therapy on SE after allowing for age in female participants is of high statistical significance (F(3,106) = 9.04, p < 0.001). The paper concludes that auricular therapy using magnetic pearls is an effective means of improving the quantity and quality of sleep in the elderly. PMID- 12568273 TI - The cardiorespiratory response and energy expenditure of Tai-Chi-Qui-Gong. AB - The purpose of this study was to evaluate the cardiorespiratory response and energy expenditure during the practice of Tai-Chi-Qui-Gong (TCQG). Forty-seven TCQG practitioners with a mean age of 60.7 +/- 9.9 years participated in this study. The cardiorespiratory responses and energy expenditure were measured by a K4 telemetry system. TCQG has 54 motions and can be divided into 3 sets. The result indicated that the exercise intensity of each motion is about 3 metabolic equivalents (MET) and the energy expenditure of each set is about 60 Kcal. The estimated intensity of TCQG in elderly individuals approximated 50% of the maximum oxygen uptake (vO2max) for men, and 60% of the maximum oxygen uptake (vO2max) for women. The findings demonstrate that TCQG is a low intensity exercise and can be prescribed as an alternative exercise program for cardiopulmonary rehabilitation. PMID- 12568274 TI - Effects of Qi-training on heart rate variability. AB - This study investigates changes in autonomic nervous function through Qi training. The power spectrum of heart rate variability (HRV) was examined in 20 sedentary healthy subjects and 20 Qi-trainees. It was found that Qi-training in healthy young subjects during controlled respiration increases the high frequency (HF) power and decreases the low frequency/high frequency (LF/HF) power ratio of HRV. These results support the hypothesis that Qi-training increases cardiac parasympathetic tone. In addition, Qi-trainees were found to have higher parasympathetic heart modulation compared with their age-matched, sedentary counterparts. This augmented HRV in Qi-trainees provides further support for long term Qi-training as a possible non-pharmacological cardio-protective maneuver. In conclusion, Qi-training may stabilize the autonomic nervous system by modulating the parasympathetic nervous system. PMID- 12568275 TI - Fabrication and evaluation of a new composite composed of tricalcium phosphate, gelatin and chi-li-saan as a bone substitute. AB - The purpose of this study was to prepare and evaluate the feasibility and biocompatibility of a new composite as a bone substitute. The new composite (GTGC) was mainly composed of tricalcium phosphate ceramics and gelatin to which chi-li-saan, a Chinese medicinal remedy was added. The GTGC composite was manually packed into cylindrical Teflon molds, dried overnight in an oven and sterilized by gamma-ray prior to use. Mature New Zealand rabbits, weighting 3-3.5 kg, underwent full-thickness excision of the parietal bone. In the experimental group, bone defects of 12 animals were filled with the GTGC composites and another 12 unreconstructed rabbits were considered as controls. Three rabbits were examined for each group in every time period at 2, 4, 8 and 12 weeks after operation. There was no evidence of adverse tissue reaction to the GTGC composite. In addition, examination with light and fluorescent microscopy revealed a significantly greater amount of new bone ingrowth in the GTGC group at the same implantion time as compared with the controls. Therefore, the GTGC composite could serve as a useful substitute when repairing bone defects. PMID- 12568276 TI - Effects of red ginseng upon postoperative immunity and survival in patients with stage III gastric cancer. AB - In this paper, we present evidence that the red ginseng powder from Panax ginseng C.A. Meyer inhibits the recurrence of AJCC stage III gastric cancer and shows immunomodulatory activities during postoperative chemotherapy, after a curative resection with D2 lymph node dissection. Flow cytometric analyses for peripheral T-lymphocyte subsets showed that the red ginseng powder restored CD4 levels to the initial preoperative values during postoperative chemotherapy. Depression of CD3 during postoperative chemotherapy was also inhibited by the red ginseng powder ingestion. This study demonstrated a five-year disease free survival and overall survival rate that was significantly higher in patients taking the red ginseng powder during postoperative chemotherapy versus control (68.2% versus 33.3%, 76.4% versus 38.5%, respectively, p < 0.05). In spite of the limitation of a small number of patients (n = 42), these findings suggest that red ginseng powder may help to improve postoperative survival in these patients. Additionally, red ginseng powder may have some immunomodulatory properties associated with CD3 and CD4 activity in patients with advanced gastric cancer during postoperative chemotherapy. PMID- 12568277 TI - Effects of hydroxyapatite in combination with far-infrared rays on spontaneous mammary tumorigenesis in SHN mice. AB - We have found that the administration of a diet containing 5% hydroxyapatite (HAP) derived from pig and cattle bones, and exposure to far-infrared rays (FIR) markedly inhibited spontaneous mammary tumorigenesis in SHN mice. Thus, the effect of combined treatment with HAP and FIR on mammary tumorigenesis was examined. The significant inhibition of tumor development by individual treatment with HAP or FIR was not enhanced by combined treatment; instead, the decrease in the inhibitory effect of HAP with age was ameliorated. Associated with this, life span was elongated and a decline in ovarian function was prevented by HAP plus FIR. Normal and preneoplastic growth of mammary glands and plasma component levels were not significantly affected by any treatment. The findings indicate that HAP and FIR have characteristics common to most natural products; in combination with other agents, they have little additive effect, when each is highly active. PMID- 12568278 TI - Neurotransmitter levels in brains of chronically Jiawey Siwu-treated rats. AB - Levels of monoamines and metabolites, excitatory amino acids, and gamma aminobutyric acid (GABA) were investigated in discrete brain areas of chronic Jiawey Siwu (JS)-treated rats. Male Sprague-Dawley rats were dosed orally for 3 months with normal saline or JS at 0.21, 1.05 or 4.2 g/kg/day. Body weights of these four groups were similar over 3 months. Most effects of JS revealed a dose dependency with levels of neurotransmitters. Levels of norepinephrine (NE) and epinephrine (EPI) in cerebral cortex; EPI, vanillylmandelic acid (VMA), dopamine (DA) and 5-hydroxytryptamine (5-HT) in medulla oblongata; DA in midbrain; NE and 5-HT in amygdala; and 5-HT in hypothalamus had decreased in JS-treated rats. 3 Methoxytyramine (3-MT) in cerebral cortex; 5-hydroxyindole-3-acetic acid (5-HIAA) in medulla oblongata; NE, 3-MT and homovanillic acid (HVA) in pons; EPI and 3-MT in midbrain; 3-MT and HVA in amygdala; 3-MT, 3,4-dihydroxyphenylacetic acid (DOPAC), HVA and 5-HIAA in cerebellum; HVA in hypothalamus; and DOPAC and HVA in hippocampus had all increased in JS-treated rats. In pons, 5-HT increased with low and decreased with high JS doses. Ratios of DA/3-MT in pons and midbrain; DA/HVA in pons and cerebellum; and 5-HT/5-HIAA in medulla oblongata, cerebellum and hypothalamus had decreased. Furthermore, aspartate (ASP) and glutamate (GLU) levels had decreased in cerebral cortex, midbrain, hypothalamus and hippocampus or amygdala, and increased in pons. GABA levels were reduced in cerebral cortex, and higher in medulla oblongata, pons, amygdala, cerebellum, hippocampus and striatum of JS-treated rats. These results indicate that the synthesis and (or) metabolism of NE, DA, EPI and 5-HT, and the levels of ASP, GLU and GABA in rat brains were differentially regionally altered by JS, which may contribute to the central manifestations of JS treatment. PMID- 12568279 TI - Tokishakuyakusan directly attenuates PACAP's luteolytic action on luteal function in the rat ovary. AB - We investigated the potential direct effects of Tokishakuyakusan (TS) on progestin [progesterone and 20alpha-hydroxyprogesterone (20alpha-OH-P)] and cyclic adenosine-3',5'-monophosphate (cAMP) production in cultured rat luteal cells. In addition, we examined whether TS regulates the inhibitory effects of pituitary adenylate cyclase-activating polypeptide (PACAP), a newly found peptide, on luteinizing hormone (LH)-stimulated progesterone production. TS significantly stimulated progesterone, but not 20alpha-OH-P, production and cAMP accumulation through 24 hours of culture. PACAP-38 significantly elevated progesterone, 20alpha-OH-P and cAMP levels at all concentrations studied. On the other hand, PACAP-38 inhibited the production of progesterone and the accumulation of cAMP enhanced by LH, while the ratio of progesterone to 20alpha OH-P was significantly decreased by PACAP-38 + LH. Concomitant treatment with TS and PACAP-38 + LH increased the ratio of progesterone to 20alpha-OH-P more than with PACAP-38 + LH. The present data have demonstrated that TS stimulates progesterone production in rat luteal cells, reconfirming our previous evidence that TS stimulates luteal steroidogenesis. The data further suggest that TS tends to attenuate PACAP's inhibition of LH-stimulated progesterone production, suggesting a luteotrophic effect within the corpus luteum. PMID- 12568280 TI - Isocupressic acid blocks progesterone production from bovine luteal cells. AB - The needles of ponderosa pine (Pinus ponderosa Laws.) were reported to induce abortions when fed to late-term pregnant beef cows in North America. An in vivo study of pregnant cows suggested that isocupressic acid (IA) was the main abortifacient isolated from needles and bark of the pine. However, the mechanism of abortifacient activity of IA is not clear yet. In a pregnant cow, the corpus luteum of the ovary helps the maintenance of pregnancy by its progesterone production. This study involved the IA extracted from the root of the Taiwan cypress (Juniperus formosana) and used a frozen-thawed bovine luteal cell culture system to investigate the action of IA on progesterone production. Thawed bovine luteal cells (1 x 10(5) cells/ml/well) in M199 medium were cultured in 24-well culture plates at 37 degrees C in a 5% CO2 incubator. Ten ml of tested drugs, IA at 1 to 1000 ng/ml and/or ovine luteinizing hormone (oLH) at 1 to 100 ng/microl or 8-bromo-cyclic adenosine monophosphate (8-Br-cAMP) with 0.1-10 mM, were added into each well. After 4 hours of incubation, the media were harvested and assayed for progesterone by an enzyme immunoassay. Progesterone production from cells was the indicator used to evaluate the action of IA. All tested doses of IA significantly inhibited progesterone production in both basal and oLH stimulating conditions. Also those dosages inhibited cyclic adenosine-3',5'- monophosphate (cAMP) stimulation, suggesting a post-cAMP mechanism is involved in the IA action. We concluded that IA can induce pregnant cows to abort partly through blocking luteal function and may be identified as a new abortifacient chemical. PMID- 12568281 TI - Effects of Orengedoku-to and Scutellariae radix extract on aqueous flare elevation induced by intravitreal IL-1alpha in pigmented rabbits. AB - We investigated the effects of Orengedoku-to (huanglian-jie-du-tang in Chinese) and Scutellariae radix extract on experimental aqueous flare elevation in pigmented rabbits. To produce the elevation of aqueous flare in rabbits, interleukin-1 alpha (IL-1alpha) was injected intravitreally. Animals were pretreated with oral administration of 150 g/day of food containing 0.07%, 0.2% or 0.7% (w/w) Orengedoku-to; or 0.02%, 0.07% or 0.2% Scutellariae radix extract for 5 days. The animals were further treated with the same food for 7 days after intravitreal injection of IL-1alpha. Aqueous flare was measured with a laser flare-cell meter. Treatment with 0.2% or 0.7% Orengedoku-to and 0.07% or 0.2% Scutellariae radix extract significantly (p < 0.05) suppressed elevation of aqueous flare induced by IL-1alpha. Thus, orengedoku-to and Scutellariae radix extract have an inhibitory effect on experimental elevation of aqueous flare induced by IL-1alpha in rabbits. PMID- 12568282 TI - A Chinese herbal medicine, fu-ling, regulates interleukin-10 production by murine spleen cells. AB - Fu-ling is one of the most widely used Chinese herbal medicines. In this study, we investigated the regulatory effect of fu-ling on interleukin-10 (IL-10) production in vivo. Mice were i.p. administered 0.1 mg to 1.0 mg fu-ling per gram body weight daily for three days. The spleen cells were isolated and assayed for both IL-10 and immunoglobulin (Ig) production. Results indicated that the mice treated with fu-ling had significantly increased spleen cell ability to secrete IL-10. Spleen cells isolated from the mice injected with either 0.1 mg or 1.0 mg fu-ling per gram body weight also showed an increase in IL-10 mRNA expression. As IL-10 is a potent differentiation factor of B-lymphocytes, the possible effect of fu-ling on Ig production was also studied. Results indicated that fu-ling significantly induced an increase in IgG and IgA secretion by spleen cells but showed no effect on IgM secretion. Thus, fu-ling may affect the function of B lymphocytes via stimulating IL-10 production. PMID- 12568283 TI - Protective effects of Wuyaoshunqisan against H2O2-induced apoptosis on hippocampal cell line HiB5. AB - Oxidative stress has been implicated in the pathogenesis of different neurodegenerative disorders. To investigate the protective effects of Wuyaoshunqisan against H2O2-induced apoptosis in the central nervous system, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) method, flow cytometric analysis, and the DNA fragmentation assay were performed on cells of the hippocampal cell line HiB5. Through the morphological and biochemical analyses, it was shown that HiB5 cells treated with H2O2 exhibit classical apoptotic features, while the occurrence of such changes is reduced in cells pre-treated with Wuyaoshunqisan prior to H2O2 exposure. PMID- 12568284 TI - The combination therapy of Ephedra herb and Loxoprofen caused gastric lesions in mice. AB - The combination therapy of a Kampo formula and an analgesic-antipyretic agent is often used for the common cold in Japan. We investigated the effect of such a combination therapy, using the Ephedra herb, which is a common ingredient of Kakkon-to and Mao-to, and Loxoprofen, a nonsteroidal anti-inflammatory drug (NSAID), on fever induced in an experimental model of mice under strong stress. The combination therapy of Ephedra herb and Loxoprofen caused gastric mucosal lesions and loss of body weight. It is considered that this combination therapy should be avoided because of its adverse effects. PMID- 12568285 TI - Toxicological interactions of Cassia senna and Nerium oleander in the diet of rats. AB - The toxic effects of diet containing 10% of C. senna L. fruits or 10% of N. oleander L. leaves or their 1:1 mixture (5% + 5%) on male Wistar rats treated for 6 weeks were investigated. Diarrhea was a prominent sign of C. senna L. toxicosis. In both phytotoxicities, there were decreases in body weight gains, inefficiency of feed utilization, dullness and enterohepatonephropathy. These findings accompanied by leukopenia and anemia were correlated with alterations of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities and concentrations of total protein, albumin, urea and other serum constituents. In both phytotoxicities, the ability of the liver to excrete bilirubin remained unchanged. Feeding the mixture of C. senna L. fruits and N. oleander L. leaves caused more serious effects and death of rats. The implications of these findings are discussed. PMID- 12568286 TI - Effect of berberine on regression of pressure-overload induced cardiac hypertrophy in rats. AB - Berberine is the basic chemical component of a Chinese herb, Coptis chinensis Franch (coptis), considered to be useful in treating some diseases of the cardiovascular system, such as hypertension and chronic heart failure (CHF). In this study, we investigate the inhibitory effect of berberine on experimental cardiac hypertrophy, which is regarded as a risk factor of CHF and other heart diseases. Forty-two male SD rats were divided into four groups: age-matched control, aortic banding model, berberine-treated group and captopril-treated group. Cardiac hypertrophy was induced by suprarenal abdominal aorta constriction (banding). The drugs were orally administered for 8 weeks starting from 4 weeks after surgery at dosage of berberine 10 mg/kg and captopril 50 mg/kg. Blood pressure (BP) was measured four times during the period of the experiment, and hemodynamic parameters, cardiac index, cell size of left ventricular myocardium and total protein of left ventricular tissue were detected 8 weeks after treatment with drugs. The data from the present study showed that: (1) The BP of the aorta banded rats was increased compared with those of the normal (p < 0.001) and the age-matched control rats (p < 0.001), and berberine showed no significant effect on it. (2) After 8 weeks of treatment with berberine, the elevated left ventricular end diastolic pressure (LVEDP) was slightly decreased compared with the aortic banded rats. Meanwhile, the maximum rates of contraction and relaxation (+/- dp/dtmax) was increased (p < 0.05) and the time to reach the point of maximum rate from beginning of contraction (t-dp/dt) was shortened (p < 0.01), indicating that the functions of heart, both contraction and relaxation, were improved. (3) Cardiac growth was inhibited by treatment with berberine. Both whole heart and left ventricular weight were notably decreased compared with the banded rats (p < 0.05 and p < 0.01). (4) The cell size of left ventricular myocardium was significantly reduced (p < 0.001) and the total protein of left ventricular tissue was slightly down-regulated by treatment with berberine. These data suggest that berberine can improve abnormal cardiac function and can prevent the development of left ventricular hypertrophy induced by pressure-overload. This indicates that it may have therapeutic potential in the treatment of CHF. PMID- 12568287 TI - Effect of tetramethylpyrazine on lipid peroxidation in streptozotocin-induced diabetic mice. AB - Oxygen free radicals have been suggested to be a contributory factor in complications of diabetes mellitus. In the present study, we investigated the effect of tetramethylpyrazine (TMP) (10, 25 and 50 mg/kg), an active component of a Chinese medicinal plant Ligusticum wallich Franch (chuanxiang), on streptozotocin (STZ)-induced diabetic male ICR mice. STZ was injected as a single daily dose (4 mg/kg i.p. in buffered citrate solution, pH 4.0) for one week. A single daily dose of TMP (10, 25 or 50 mg/kg i.p.) was administered for 2 weeks to the STZ-induced diabetic mice immediately following 1 week STZ induction. Plasma glucose and serum blood urea nitrogen (BUN) concentrations were estimated at the time of sacrifice. Malonialdehyde (MDA) formation was measured from the liver and kidney tissues. TMP dose dependently inhibited glucose concentration and BUN elevation. TMP also remarkably reduced the degree of lipoperoxidation. Our results indicate that TMP may be an effective agent for treatment of diabetes mellitus complications with oxidative stress. PMID- 12568288 TI - The evaluation of reno- and hepatoprotective effects of huai-shan-yao (Rhizome Dioscoreae). AB - Huai-shan-yao (Chinese yam; Rhizome Dioscoreae) is a common food in China. In the present study, we evaluated the protective effects of the crude extract of huai shan-yao on acute kidney and liver injuries in rats induced by ethanol. Results of pharmacological, biochemical and pathologic observations all showed that rats treated with the extract of huai-shan-yao had decreased damage in renal tubules as well as decreased inflammation in the central vein and necrosis in the liver tissue. PMID- 12568289 TI - Effect of Kampo herbal medicines on murine water metabolism in a microgravity environment. AB - To determine the possibility of new applications of Oriental medicines, we examined the changes in water metabolism of mice that underwent microgravity and were treated with Kampo medicines. Male ICR mice were used in this experiment. Eight extracts of Kampo herbal medicines were dissolved in water and added to the drinking water administered to mice at 1 g/kg body weight for two days. The microgravity experiment was performed at the Japan Microgravity Center. We used a drop-shaft type microgravity experimental system with a free fall of 490 m. Before the drop, 7 ml of physiological saline was injected intraperitoneally. Under fasting and dehydration, body weights were measured and loss of body weight was calculated as urine. Blood samples were collected 24 hours after the microgravity experiment, and the antidiuretic hormone (ADH) in plasma related to water metabolism was measured by the radioimmunoassay (RIA) method. Heat shock protein in the spleen was measured by the enzyme-linked immunosolvent assay (ELISA) method. In the Hachimi-jio-gan and Hochu-ekki-to groups in microgravity, a decrease of urine was observed, which significantly suppressed the increase of ADH due to microgravity. Hachimi-jio-gan reduced the content of heat shock protein (HSP) 70 in the spleen. It is suggested that Hachimi-jio-gan and Hochu ekki-to could be used as water metabolism adjustment reagents in a space environment. Furthermore, it is suggested that Hachimi-jio-gan could ease the stresses caused by microgravity. The physiological changes resulting from a microgravity environment are serious problems for space flight. Pre-treatment with Kampo medicines is expected to prevent, ease and treat these problems. PMID- 12568290 TI - The in vitro inhibitory effects of crude extracts of traditional Chinese herbs on 3-hydroxy-3-methylglutaryl-coenzyme A reductase on Vero cells. AB - Cardiovascular disease is still the leading cause of death in Western countries. Epidemiological studies have shown that hypercholesterolemia is a major risk factor for coronary artery disease. Clinical trials of lipid lowering therapy with 3-hydroxy-3-methylglutaryl coenzyme A (HMG Co-A) reductase inhibitor have been shown to decrease coronary events and mortality. Flavonoids are polyphenolic natural antioxidants occurring in natural products such as traditional Chinese herbs, fruits and beverages such as tea and wine. The aim of this study was to evaluate the effects of crude extracts from traditional Chinese herbs on HMG Co-A reductase. The methods for analysis of specific inhibitors of mevalonate biosynthesis have been well-established by using Vero cells, a cell line obtained from kidneys of African green monkeys. Crude extracts from different traditional Chinese herbs were dissolved in 1% Dulbecco's modified Eagle's medium and incubated with Vero cells with or without the addition of 1 mM mevalonate or 5 mM sodium acetate for 24 hours in order to observe cell growth. Pravastatin, a specific HMG Co-A reductase inhibitor, was used as a positive control which inhibits Vero cells growth effectively and cell growth inhibition was reversible after 1 mM mevalonate. Among 100 traditional Chinese herbs used for the study, only two herbs: Curcuma zedoaria Roscoe and Poncirus trifoliata Raf. showed significant growth inhibition of Vero cells. This study shows that some crude extracts isolated from traditional medicinal herbs were effective HMG Co-A reductase inhibitors which might be developed into new hypocholesterolemic agents. PMID- 12568291 TI - A hospital-based study on the use of alternative medicine in patients with chronic liver and gastrointestinal diseases. AB - To determine the extent to which Taiwanese patients use alternative medicine, we interviewed 500 consecutive patients with chronic liver and gastrointestinal disorders at an outpatient-service. Forty-two patients were excluded due to incomplete data. The percentages of patients with chronic liver (102/269, 37.9%) and gastrointestinal (74/189, 39.2%) diseases using alternative medicine were not significantly different (p = 0.70). The patients who used alternative medicine were not statistically different in gender (p = 0.37), age (p = 0.59), education level (p = 0.83), family income (p = 0.90), or occupation (p = 0.72). Only 36% (64/176) of patients informed their doctors of their use of alternative medicine. The kinds of alternative medicine used by the 176 patients included: Chinese/herbal medicine, 169 (96%); acupuncture, 31 (18%); nutritional supplements, 22 (13%); chiropractic, 17 (10%); scratching, 14 (8%); Qigong, 13 (7%); cupping, 13 (7%); and incense ash, 3 (2%). Sixty-six percent (111/169) of patients used Chinese/herbal medicine in addition to Western allopathic medicine. Only 11% (19/169) of them believed that Chinese/herbal medicine had side effects. Our study indicates the use of alternative medicine occurs across all demographic groups in one-third of patients with chronic liver and gastrointestinal diseases at a major general hospital in Taipei. We suggest that the doctors question all patients for history of alternative therapy use. PMID- 12568292 TI - Anti-diabetic and anti-obese effects of ginseng berry extract: comparison between intraperitoneal and oral administrations. PMID- 12568293 TI - Recent developments in basic and clinical hematopoietic stem cell transplantation. PMID- 12568294 TI - Review of "minitransplantation": nonmyeloablative allogeneic hematopoietic stem cell transplantation. AB - Nonmyeloablative conditioning regimens for allogeneic hematopoietic stem cell transplantation (HSCT) have been developed over the past few years as important alternatives to conventional myeloablative conditioning regimens for older or medically unfit patients with hematologic malignancies, as well as for patients with certain nonmalignant hematologic diseases or renal cell cancer. This review summarizes the biological background, current clinical applications, and indications for this novel treatment approach for treating hematologic malignancies. Historically, allogeneic HSCT has been based on the use of cytotoxic and myeloablative chemotherapy and radiotherapy conditioning regimens that are intended both to eradicate malignancy and to eliminate host hematopoiesis and immune cells. Such a regimen was followed by the infusion of histocompatible donor marrow or peripheral blood stem cells to rescue hematopoiesis. For older patients or for those who had previously been treated with intensive chemotherapy or radiotherapy, the toxicity of myeloablative conditioning was prohibitive. Although most hematologic malignancies occur in older patients, these patients had not been previously eligible for the potentially curative therapy offered by allogeneic HSCT. Based in large part on preclinical studies with the dog model of HSCT and on an improved understanding of the mechanisms for controlling immune modulation, successful development of nonmyeloablative conditioning regimens for clinical use has occurred. Clear evidence of a therapeutic graft-versus-tumor effect mediated by allogeneic T cells prompted an exploration for HSCT regimens that rely solely on nonmyeloablative immunosuppression to facilitate allogeneic engraftment. In lieu of intensive chemoradiotherapy before transplantation, engrafted donor T-cells are used to accomplish the task of eradicating the host's malignant cells. We review the updated results of an ongoing multicenter study to investigate the safety and efficacy of nonmyeloablative HSCT using a regimen of 2 Gy total body irradiation in patients with advanced hematologic malignancies who were ineligible for conventional myeloablative conditioning. In addition, we review the results of reduced-intensity HSCT trials from other transplantation centers. PMID- 12568295 TI - New strategies for prevention and treatment of graft-versus-host disease and for induction of graft-versus-leukemia effects. AB - Graft-versus-host disease (GVHD) continues to be a problem in allogeneic hemopoietic stem cell transplantation; however, our understanding of the basic pathophysiology of GVHD has improved. Although not all data obtained from murine or other animal models can be extrapolated to the clinic, there are leads that deserve to be pursued. The skin, intestinal tract, and liver are the 3 major target organs of GVHD and share the feature of presenting a barrier to the "environment" of the host. There is evidence that the damage inflicted to these organs, the epithelial and endothelial cells in particular, by the conditioning regimen causes a release of various cytokines and a penetration of endotoxin into the systemic circulation. According to these observations, the nonimmunologic aspects of GVHD have been likened to an inflammatory process. If this characterization is valid, blocking these nonspecific inflammatory changes would ameliorate GVHD without interfering with the graft-versus-leukemia (GVL) reaction. In fact, one study has shown a substantial amelioration of GVHD with a molecule that directly blocks endotoxin. Clinical data also suggest that patients with organ dysfunction early after transplantation that is presumed to be treatment related may benefit from preemptive interventions aimed at controlling GVHD. Furthermore, there is growing evidence that the mechanisms involved in GVHD may differ from organ to organ (for example, Fas/Fas-ligand interactions in the liver versus tumor necrosis factor alpha/receptor interactions in the intestinal tract), and from a therapeutic point of view, the time of onset of clinical GVHD may be important in choosing the appropriate therapy. Thus, combinations of interventions chosen and timed appropriately may be more effective in preventing and managing GVHD than are the standard across-the-board approaches that have been used so far. Such a strategy may also be successful in maintaining a GVL effect and possibly in incorporating direct antileukemic therapy, such as the use of cytotoxic T-cells directed at minor histocompatibility antigens, without increasing the risk of GVHD. The development of nonmyeloablative conditioning regimens and the observations on GVHD kinetics and the progression or eradication of leukemia with that strategy are likely to add new insights into how one can optimally combine various modalities to achieve engraftment, prevent GVHD, and at the same time maintain a GVL effect. PMID- 12568296 TI - Role of immunotherapy in stem cell transplantation. AB - Relapse of the underlying malignancy continues to be a major problem after both autologous and allogeneic stem cell transplantation. Over the years, it has been recognized that immune-mediated graft-versus-tumor effects are crucially involved in eliminating minimal disease and controlling its recurrence after stem cell transplantation. This recognition has led to a number of studies that have attempted to stimulate a cellular immune response in the recipient, especially after allogeneic transplantation. Immunotherapy after autologous transplantation has to take into consideration the fact that patients' immune cells frequently are compromised and tolerance to the host tumor may have developed. Hence, trials involving the administration of cytokines (such as with interleukin and interferon) have shown limited benefits. This situation is different for allogeneic transplantation for which the infusion of donor lymphocytes has shown disease regression, especially in patients with chronic leukemias. However, such treatment is effective only if the patient has limited disease, and severe graft versus-host disease frequently has to be accepted as a complication. This fact has led investigators to pursue the generation of specific lymphocytes that can recognize tumor antigens but not necessarily induce graft-versus-host disease. Such studies are in the early stages, and although some promising results have been observed, it is unclear at this point if the antitumor effect can be separated sufficiently from the graft-versus-host disease mediated by allogeneic lymphocytes. More recently, it has been shown that natural killer (NK) cells can have an antitumor effect in myeloid malignancies, particularly if the cells are allogeneic and do not recognize self-HLA antigens. At this point, it appears that engineered T-lymphocytes and allogeneic NK cells may be useful in preventing or treating relapse after allogeneic transplantation. It remains to be seen if such novel cellular therapies can also be implemented after autologous transplantation via the use of engineered allogeneic immune cells. PMID- 12568297 TI - Genetic control of stem cells: implications for aging. AB - Stem cells are currently at the center of both controversy and notoriety. The harvest of human embryonic or fetal stem cells, at least with methods available now, necessarily involves the sacrifice of the embryo or fetus. This critical step in the procurement of stem cells has stimulated intense discussion at all levels of academia, government, and society in general. What societal benefits, if any, justify such a strategy for obtaining these stem cells? In other species it has been possible to generate virtually all cell types found in adult organs from embryonic stem cells. This ability has opened endless clinical possibilities for tissue and organ replacement through the transplantation of cells derived from embryonic stem cells. Luckily, there may be an alternative to this ethical dilemma. It is becoming increasingly clear that stem cells exist in many, if not all, adult tissues. Adult stem cells normally replenish tissue cells lost through the wear and tear of aging or damage from injury or disease. With the proper coaxing in tissue culture and when transplanted, these stem cells may regenerate the full repertoire of organotypic cells and thus may therapeutically regenerate tissues in vivo in much the same way as embryonic stem cells do. For several reasons, the best-studied stem cells are those of the blood-forming system. Mature blood cells generally have short functional life spans, usually measured in days, and therefore require replenishment at a steady pace throughout one's lifetime. Stem cells are intimately involved in this renewal and, because of the relative ease of access to the bone marrow, stem cells have been well studied. Second, bone marrow transplantation following radiation or high-dose chemotherapy in the treatment of cancer has fostered research on the basic biology and therapeutic uses of hematopoietic stem cells over the more than 30 years stem cell transplantation has been used clinically. It is my aim to review what is known about the genes controlling hematopoietic stem cell function. Identifying, and ultimately manipulating, the genes that regulate stem cell number, replication rate, and self-renewal capacity may have important clinical benefits. I discuss evidence suggesting that the characterization of least some of these stem cell genes will shed light on mechanisms important in the aging process. I advance the hypothesis that stem cells accumulate cellular damage during aging that diminishes their developmental potency and ability to replenish blood cells, particularly after hematopoietic stress. In this view, the impaired function of stem cells in hematopoietic and in other self-renewing tissues limits the longevity of animals, and perhaps of humans. PMID- 12568299 TI - Frequencies of the hereditary hemochromatosis allele in different populations. Comparison of previous phenotypic methods and novel genotypic methods. AB - AIM: The frequencies of the hereditary hemochromatosis allele were compared for different populations assessed by previous phenotypic methods and the present genotypic methods. METHODS: From a literature survey, the calculated hemochromatosis allele frequencies from 16 studies using phenotypic biochemical markers (threshold levels for transferrin saturation [range, 46%-70%] and serum ferritin [range, 164-700 microg/L]) were compared with allele frequencies of the Cys282Tyr mutation of the hemochromatosis gene reported in 19 genotypic studies. RESULTS: Calculated phenotypic allele frequencies are high in Scandinavia: Iceland, 6.1% to 7.4%; Norway, 5.8%; central Sweden, 6.3% to 6.9%; Denmark, 6.1%. Frequencies are similarly high in Wales, Canada, Utah, South Africa, and Australia (range, 5.2%-9.8%). Frequencies are low in Finland (1.9%) and northern Italy (4.5%). Genotypic allele frequencies of the Cys282Tyr mutation are likewise high in Scandinavia. Frequencies are high in the United Kingdom and northern France and low in Finland, central Germany, northern Italy, and Greece. The phenotypic-genotypic ratios of the hemochromatosis homozygosity frequencies for the same geographic area were calculated. A ratio of 1.0 indicates that the 2 methods give similar results. In 3 studies, the ratio was above 1.0, the highest ratio of 1.67 being reported from Italy. In most studies the ratio was slightly below 1.0 (0.71-0.97). The lowest ratio was found in Finland (0.33). CONCLUSION: In most studies there was good agreement between the hemochromatosis allele frequencies determined by phenotypic and genotypic methods. A high ratio (northern Italy) may indicate that phenotypic selection criteria were too loose and/or that causes of iron overload other than the Cys282Tyr mutation are frequent in the region. A low ratio (in Finland) may indicate phenotypic selection criteria that were too stringent and/or a low penetration rate of the mutation. PMID- 12568298 TI - Hodgkin's lymphoma and CD30 signal transduction. AB - Advances in molecular biology have shed light on the biological basis of Hodgkin's lymphoma (HL). Knowledge of the biological basis has enabled us to understand that most Hodgkin and Reed-Sternberg (H-RS) cells are derived from germinal center B-cells and constitutive nuclear factor kappaB (NF-kappaB) activation is a common molecular feature. Molecular mechanisms responsible for constitutive NF-kappaB activation, Epstein Barr virus latent membrane protein 1, and defective IkappaBalpha and IkappaB kinase activation have been clarified in the past several years. A recent study revealed the biological link between 2 characteristic features of H-RS cells: CD30 overexpression and constitutive NF kappaB activation. Ligand-independent signaling by overexpressed CD3O was shown to be a common mechanism that induced constitutive NF-kappaB activation in these cells. These results suggest the self-growth-promoting potential of H-RS cells and redefine the biology of HL composed of H-RS cells and lymphocytes. PMID- 12568300 TI - Recovery of normal hematopoiesis after severe bone marrow aplasia induced by interferon-alpha in a patient with chronic myelogenous leukemia. AB - We describe an interesting case of a patient with chronic myelogenous leukemia (CML) who developed sustained severe bone marrow aplasia after 2 years and 11 months of interferon-alpha (IFN-alpha) therapy but demonstrated recovery of normal hematopoiesis when treated with immunosuppressive therapy with granulocyte colony stimulating factor (G-CSF). Administration of G-CSF resulted in a partial recovery of hematopoiesis, and after starting immunosuppressive therapy, the patient was no longer dependent on blood transfusions. Moreover, her bone marrow had no Philadelphia chromosome-positive clones. According to the results of the present case, bone marrow recovery can be achieved with immunosuppressive therapy and a fatal outcome avoided, even in CML patients suffering from sustained bone marrow aplasia during IFN-alpha treatment. PMID- 12568302 TI - Cyclic platelet and leukocyte count oscillation in chronic myelocytic leukemia regulated by the negative feedback of transforming growth factor beta. AB - We report a case of chronic myelocytic leukemia (CML) with cyclic oscillation of platelet and leukocyte counts and attempt to elucidate the oscillatory mechanism from the standpoint of cytokine regulation of hematopoiesis. A 57-year-old woman with a diagnosis of CML exhibited platelet and white blood cell (WBC) count fluctuations of a cyclic nature. The average duration of the cycles was about 8 weeks. The patient suffered from headache, fatigue, and malaise at the peak of the cycle. The peak thrombopoietin concentration in peripheral blood coincided with a period of decrease in platelet numbers. The change in transforming growth factor beta (TGF-beta) level paralleled that of the platelet numbers. A progenitor cell assay revealed the suppression of trilineage colony formation in the presence of plasma from the blood cell peak point, and this suppression was completely blocked when the plasma was incubated with an anti-TGF-beta antibody. From these findings, we concluded that the cyclic oscillation of the platelet, WBC, and reticulocyte counts had been induced by excess negative feedback to megakaryopoiesis by TGF-beta. PMID- 12568301 TI - Antiapoptotic effect of granulocyte colony-stimulating factor, granulocyte macrophage colony-stimulating factor, and cyclic AMP on human neutrophils: protein synthesis-dependent and protein synthesis-independent mechanisms and the role of the Janus kinase-STAT pathway. AB - Spontaneous neutrophil apoptosis during culture was delayed by granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), or dibutyryl cyclic adenosine monophosphate (cAMP), whereas apoptosis was accelerated by cycloheximide or actinomycin D. G-CSF-mediated antiapoptosis was completely abolished by cycloheximide or actinomycin D, whereas GM-CSF-mediated antiapoptosis was not completely abolished by these inhibitors. Antiapoptosis induced by dibutyryl-cAMP was highly resistant to cycloheximide, and that induced by benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone was unaffected by cycloheximide. G-CSF- and GM-CSF-mediated antiapoptosis and phosphorylation of signal transducer and activator of transcription 3 (STAT3) and STAT5 were inhibited by AG490, an inhibitor of Janus kinase. The level of Mcl-1 protein was not associated with neutrophil apoptosis. The results suggest that (a) neutrophil survival in the resting state is primarily regulated by the constitutive synthesis of antiapoptotic proteins; (b) the prevention of spontaneous apoptosis is mediated through the protein synthesis-dependent and/or protein synthesis independent mechanisms according to the stimuli used; and (c) the Janus kinase STAT pathway is involved in G-CSF- and GM-CSF-mediated antiapoptosis. PMID- 12568303 TI - Effect of granulocyte colony-stimulating factor on bone metabolism during peripheral blood stem cell mobilization. AB - Granulocyte colony-stimulating factor (G-CSF) has been shown to affect the biochemical markers of bone metabolism, including serum bone alkaline phosphatase (BALP), serum osteocalcin, and urine deoxypyridinoline. To determine the association between bone resorption and formation and the G-CSF-induced mobilization of peripheral blood stem cells (PBSC), we examined these markers during mobilization in 19 healthy donors. The average (+/- SEM) serum BALP level before treatment was 81.6 +/- 17.0 IU/dL, and the level increased significantly to 117.7 +/- 15.8 IU/dL on day 5 of G-CSF administration (P < .0001). The urine deoxypyridinoline level before treatment was 12.3 +/- 2.4 nmol/mmol creatinine, and this level also increased significantly to 19.4 +/- 3.0 nmol/mmol creatinine on day 5 of G-CSF administration (P < .0001). In contrast, the average level of serum osteocalcin significantly decreased from 8.07 +/- 2.88 ng/mL to 1.53 +/- 0.18 ng/mL on day 5 (P = .0353). During G-CSF administration, we also studied the serum levels of various cytokines (IL-1beta, osteoclastogenesis inhibitory factor [OCIF], IL-6, tumor necrosis factor alpha, transforming growth factor beta, interferon-gamma, macrophage colony-stimulating factor) related to bone metabolism. Only the kinetics of OCIF were significantly affected. The serum level of OCIF increased immediately after the start of G-CSF administration and remained high during G-CSF administration. These results demonstrate that high dose G-CSF affects bone metabolism and that OCIF may play a role in bone metabolism. Consistent with the notion that G-CSF affects bone metabolism, a significant correlation was observed between CD34+ cell yield and the increase in urine deoxypyridinoline but not for the changes in serum BALP and osteocalcin levels. This result suggests that bone resorption is either directly or indirectly related to the mobilization of PBSC by G-CSF. PMID- 12568304 TI - Successful cord blood stem cell transplantation for myelodysplastic syndrome with Behcet disease. AB - A 10-year-old girl who had been treated as an outpatient for neurofibromatosis type 1 was admitted with fever, 10% weight loss, and abdominal pain that had persisted for 1 month. The clinical manifestations and the results of endoscopy led to the diagnosis of intestinal Behcet disease. Blood cell dysplasia appeared during the hospitalization. The patient subsequently developed blast cells, and the diagnosis of myelodysplastic syndrome (MDS) was made. Behcet disease was first controlled with steroids and diet followed by chemotherapy and cord blood stem cell transplantation (SCT). Both the MDS and Behcet disease went into remission after transplantation. The use of hematopoietic SCT has the potential not only to cure but also to help explicate the mechanism of Behcet disease. PMID- 12568305 TI - Granisetron plus dexamethasone versus granisetron alone in the prevention of vomiting induced by conditioning for stem cell transplantation: a prospective randomized study. AB - This prospective randomized study compared the efficacy and toxicity of granisetron and dexamethasone to those of granisetron alone for antiemetic control in patients receiving high-dose chemotherapy with or without total body irradiation (TBI) for stem cell transplantation. Patients were divided randomly into 2 groups. Groups received granisetron twice daily at a dose of 40 microg/kg with or without 4 mg dexamethasone (GS group and G group, respectively), starting 30 minutes before each dose of chemotherapeutic agent or TBI, or 12 hours after the first dose if TBI or a drug was given once a day. Fifty patients were evaluated for the analysis. During the first 24 hours of conditioning, 23 of 25 patients (92.0%) in the GS group achieved complete control of emesis (no emetic episodes over the course of a day), compared with 72.0% in the G group (P = .06). For patients receiving TBI on the first day of conditioning, complete emetic control was achieved in all patients (100.0%) in the GS group, compared with 63.2% in the G group (P = .02). The same degree of emetic control was maintained throughout the conditioning period in 38.8% of the GS group and 29.9% of the G group (P = .10). Adverse reactions were observed more frequently in the GS group (68.0% versus 5.0% in the G group). These reactions included insomnia, headache, flushing, and hyperglycemia. None of the events were serious. We conclude that granisetron with dexamethasone seems superior to granisetron alone for the prevention of emesis resulting from the conditioning regimen; however, the more frequent side effects may limit the wide use of this combination. PMID- 12568306 TI - Fluorescence in situ hybridization detection of AML-1/ETO rearrangement in a case of acute myelogenous leukemia with complicated cytogenetic abnormalities. PMID- 12568307 TI - Thrombocytopenia induced by imatinib mesylate (Glivec) in patients with chronic myelogenous leukemia: is 400 mg daily of imatinib mesylate an optimal starting dose for Japanese patients? PMID- 12568308 TI - The number of people in Japan with coagulation disorders: 2001 update. PMID- 12568309 TI - Low-dose antithymocyte globulin for treatment of steroid-pulse-resistant acute graft-versus-host disease. PMID- 12568310 TI - Antitumor effect of G3139 Bcl-2 antisense oligonucleotide is independent of its immune stimulation by CpG motifs in SCID mice. AB - The Bcl-2 antisense oligonucleotide (AS-ODN) G3139 chemosensitizes human malignancies by downregulating the antiapoptotic protein Bcl-2. Because G3139 contains two potential immunostimulatory CpG motifs, we asked if immune stimulation contributes to the antitumor activity observed previously. 5' Methylation of cytosines in CpG motifs abrogates immune stimulation by oligonucleotides. We, therefore, studied the antitumor and immunostimulatory potential of G3139 vs. an identical oligonucleotide, except for methylation of cytosines in the two CpG motifs (G4232). In a human melanoma SCID mouse xenotransplantation model, G3139 or G4232 was administered by continuous subcutaneous (s.c.) or bolus intraperitoneal (i.p.) infusion. Both G3139 and G4232 significantly reduced tumor growth by about one third relative to the saline-treated group. Furthermore, we noted a similar downregulation of Bcl-2 expression and increase in tumor cell apoptosis caused by G3139 and G4232 compared with saline controls. However, mice treated with G3139 had a pronounced increase in spleen weight and interleukin-12 (IL-12) plasma levels relative to mice treated with either G4232 or saline. Splenomegaly and elevated IL-12 plasma levels suggest that G3139 can be immunostimulatory. However, there is clear evidence that the antitumor effect of G3139 in this model appears to be a Bcl-2 antisense effect that is independent of immune stimulation, as G3139 and its immune-silent counterpart G4232 caused similar tumor suppression and apoptosis induction. PMID- 12568311 TI - Sequence dependence of C5-propynyl-dU,dC-phosphorothioate oligonucleotide inhibition of the human IGF-I receptor: mRNA, protein, and cell growth. AB - Human keratinocytes are highly responsive to mitogenic and antiapoptotic signaling by the insulin-like growth factor-I receptor (IGF-IR). IGF-IR hyperstimulation is a feature of hyperplastic skin conditions, making the IGF-IR an appealing target for antisense therapeutic intervention. In this study, we used a C5-propynyl-dU,dC-phosphorothioate oligo-2'-deoxyribonucleotide antisense 15-mer to the human IGF-IR mRNA, along with liposome transfection, to inhibit IGF IR activity in a human keratinocyte cell line and demonstrated potent inhibition of cell growth despite the presence of serum. To investigate the sequence specificity of these effects and to establish the concentration range over which a purely antisense effect could be demonstrated, we introduced 1, 2, 4, 8, and 15 base mismatches into the oligonucleotide and analyzed changes in inhibitory efficacy. In the 10-30 nM concentration range, the introduction of 1 and 2 mismatches into the middle of the 15-mer only modestly affected inhibitory efficacy, whereas >4 mismatches profoundly reduced mRNA, protein, and growth inhibitory effects. From these results, we conclude that (1) sequence-specific antisense inhibition of IGF-IR activity in keratinocytes is achievable, (2) potent anti-IGF-IR antisense inhibition can be achieved in vitro at concentrations as low as 10 nM, and (3) a sequence-dependent mechanism is likely to underpin the observed in vivo therapeutic effects (Wraight et al. Nat. Biotechnol. 2000;18:521) of these antisense oligonucleotides (AS-ODN) in cutaneous hyperplastic disorders, such as psoriasis. PMID- 12568312 TI - Bromodeoxyuridine-labeled oligonucleotides as tools for oligonucleotide uptake studies. AB - The mechanisms by which various oligonucleotides (ODNs) and their analogs enter cells are not fully understood. A common technique used in studies on cellular uptake of ODNs is their conjugation with fluorochromes. However, fluorescently labeled ODNs may vary from the parent compounds in charge and hydrophilicity, and they may interact differently with some components of cellular membranes. In this report, we present an alternative method based on the immunofluorescent detection of ODNs with incorporated 5-bromo-2'-deoxyuridine (BrdUrd). Localization of BrdUrd-modified ODNs has been achieved using FITC-labeled anti-BrdUrd antibodies. This technique allowed determination of the differences in cellular uptake of phosphodiester (PO) and phosphorothioate (PS) ODNs and their derivatives conjugated with cholesterol and menthol. The immunocytochemical method also has shown that the cellular uptake of some ODNs may be influenced by specific sequences that are responsible for the formation of higher-order structures. PMID- 12568313 TI - Cytosine residues influence kidney accumulations of 99mTc-labeled morpholino oligomers. AB - Watson-Crick pairing between complementary oligomers is proving to be an effective means for rapidly directing radioisotopes specifically to the exterior surface of cancer cells in vivo. In such pretargeting applications, it is highly desirable that the excess of isotopically labeled oligomers, which do not bind to the cancer cells, be rapidly cleared from the body. In this context, understanding the influence of chain length and base sequence of the radiolabeled oligomers is critical. We had earlier determined that the kidneys are the principal targets of short-chain radiolabeled morpholino oligomers (MORFs). To explain these observations, MORFs consisting of uniform cytosines (Cs), uniform thymines (Ts), uniform adenines (As), and uniform AAG repeat were labeled with technetium-99m (99mTc) and studied in normal mice. In a limited investigation of the influence of oligomer backbone, a 20-mer MORF (MORF20) with a base sequence rich in Cs was compared with a phosphoromonothioate DNA (S-DNA20) of the same sequence. The in vivo behavior of the labeled MORFs was nearly identical in all organs, with the exception of kidneys. The kidney accumulations were about 25- to 80-fold higher for the uniform Cs relative to the other three uniform MORFs at 3 hours. The S-DNA20 rich in Cs showed only modest kidney accumulations compared with the equivalent MORF20, presumably because of preferential clearance of the S DNA20 through the liver. Urine analysis showed no evidence of intact labeled S DNA20 in contrast to fully intact labeled MORF20. We conclude that the high kidney levels observed by us previously for MORFs are most likely due largely to the C residues in the base sequence. In the case of S-DNAs, this phenomenon is partly disguised by the increased hepatic excretion and degradation. These results show that the base sequences of MORFs, and probably other oligomers as well, are an important determinant of biodistribution. PMID- 12568314 TI - Inhibition of CD4 expression by antisense oligonucleotides in PMA-treated lymphocytes. AB - To decrease CD4 expression on T helper (Th) lymphocyte surface, antisense oligonucleotides (AS-ODNs), delivered by the cationic liposome DOTAP, were assayed in vitro on rat spleen lymphocytes. Four 21-mer ODNs (AS-CD4-1, AS-CD4-2, AS-CD4-3, and AS-CD4-4) directed against the translation start region of the cd4 gene were designed. AS-CD4-1 was phosphorothioate (PS)-modified in each base, and the other three were PS-modified at both ends and in the internal pyrimidine residues. Four ODN controls (fully PS-modified ODN-A and partially modified ODN B, ODN-C, and ODN-D) were also assayed. CD4 resynthesis was stimulated by treatment with phorbol 12-myristate 13-acetate (PMA) at the same time as the incubations with the ODN. After 24 hours of treatment, CD4 expression was measured by immunofluorescence staining and flow cytometry. CD4 reexpression in rat PMA-treated lymphocytes was counteracted by 40% by means of AS-CD4-2 and AS CD4-4 treatments. On the other hand, AS-CD4-3 produced only 20% inhibition, similar to that produced by ODN-B, and AS-CD4-1 did not have any significant effect compared with control ODNs. Both AS-CD4-2 and AS-CD4-4 decreased CD4 mRNA, as determined by RT-PCR, and in addition, they did not affect the expression of other surface lymphocyte molecules. Inhibition of surface CD4 expression remained at least 72 hours. The addition of both AS-ODNs did not further increase the effect obtained separately by each AS-ODN. Treatment of rat PMA-lymphocytes with two concentrations of AS-CD4-2 and AS-CD4-4 added 24 hours apart did not further decrease CD4 expression. In summary, AS-CD4-2 and AS-CD4-4 could constitute a good strategy to inhibit CD4 expression on Th lymphocytes and modulate their function. PMID- 12568315 TI - The effectiveness of double-stranded short inhibitory RNAs (siRNAs) may depend on the method of transfection. AB - RNA interference (RNAi) is a recently described powerful experimental tool that can cause sequence-specific gene silencing, thereby facilitating functional analysis of gene function. Consequently, we became interested in using RNAi to determine the function of aberrantly expressed ErbB3 in the KAS-6/1 human myeloma cell line. Despite the wealth of information available on the use of RNAi, dsRNA target design, and the transfection of dsRNA in vitro, little information is available for transfecting dsRNA into nonadherent cells from any species. In the present study, we report that gene silencing of ErbB3 was not observed in myeloma cells when dsRNA targeting ErbB3 was introduced using conventional transfection agents and protocols that have proved successful for several adherent cell lines. Silencing of ErbB3, however, was observed in T47D cells, an adherent breast carcinoma cell line, using the same transfection methods, indicating that our target sequence was functional for gene silencing of ErbB3. Interestingly, ErbB3 was silenced in myeloma cells when the dsRNA target was introduced by electroporation. Thus, our studies illustrate the striking dependence of dsRNA mediated gene silencing in some cells on the methods of dsRNA transfection. PMID- 12568316 TI - Histologically classified venous angiomas of the brain: a controversy. AB - The term "venous angioma" (VA) usually refers to a developmental venous anomaly (DVA). However, a group of vascular malformations called VAs shows no venous abnormalities on angiography. The clinical and histological features of histologically classified VAs were studied in eight patients who presented with hemorrhage or seizures to reevaluate these venous anomalies. Angiography showed no venous abnormalities in six patients. Histological study included immunostaining for smooth muscle actin and glial fibrillary acidic protein. Surgical specimens of 10 cases of cavernous angiomas, 10 cases of arteriovenous malformations, and two cases of capillary telangiectasias were studied to compare these types of VAs. Angiographically occult VAs were surgically removed safely, whereas removal of DVAs was complicated by brain swelling and hemorrhagic infarction of the brain. Histological examination found angiographically occult VAs contained malformed and compactly arranged vessels with partly degenerated walls, whereas DVAs had dilated thin-walled vessels that were diffusely distributed in the normal white matter. This study of our cases and a review of the reported cases of VAs suggests that two different clinical and pathological entities are commonly categorized as "VA," angiographically occult VAs and DVAs. These two entities should be carefully distinguished. PMID- 12568317 TI - Detection of chromosomal imbalances in spinal meningiomas by comparative genomic hybridization. AB - Little is known about genetic mutations during the malignant progression of spinal meningiomas. This study investigated genomic changes across the entire genome in spinal meningioma samples to determine possible mechanism(s) of tumorigenesis. Paraffin-embedded tissue sections of 16 spinal meningiomas were analyzed by the comparative genomic hybridization (CGH) technique. Lymphocytes of the patients were evaluated as controls. Genomic change was detected in 11 samples. Complete or partial loss of chromosome 22 was the most commonly seen abnormality in eight cases. Chromosome losses on 1p, 9p, and 10q and gains on 5p and 17q were the other abnormalities. These changes are all frequently seen in meningiomas, but are mostly specific to atypical and anaplastic meningiomas. However, in the present study, copy number changes on chromosomes 9p (3 samples), 17q (2 samples), and 1p (2 samples) were seen even in the benign tumors. Our results suggest that in addition to the neurofibromatosis type 2 tumor suppressor gene, other cancer-related genes located on 1p, 9p, 10q, and 17q might be involved in the etiology of spinal meningiomas. PMID- 12568318 TI - Caspase activation in neuronal and glial apoptosis following spinal cord injury in mice. AB - The involvement of caspases in apoptosis after spinal cord injury (SCI) was investigated in adult mouse spinal cord after contusion. Sections of spinal cord were processed for staining 7 days after SCI with the fluorescent dye Hoechst 33342, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL), and immunostaining with an antibody (CM1) recognizing activated caspase-3. Caspase-3- and caspase-8-like enzyme activities were measured colorimetrically at 8 hours to 7 days after SCI using the specific substrates Asp-Glu-Val-Asp-p-nitroanilide and Ile-Glu-Thr-Asp-p-nitroanilide, respectively. Hoechst 33342 staining showed small, bright areas in fragmented nuclei. Double labeling with TUNEL plus immunostaining with cell type-specific markers identified TUNEL-positive neurons stained by anti-neuronal nuclear protein/neurons antibody, and TUNEL-positive oligodendrocytes stained by anti cyclic nucleotide 3'-phosphohydrolase antibody. Double labeling with CM1 and cell type specific markers similarly identified CM1-positive neurons and oligodendrocytes. Caspase-8-like enzyme activity was increased significantly on days 3 and 7 (p < 0.01), whereas caspase-3-like activity increased on day 7 (p < 0.01). Intraventricular injection of a nonspecific tetrapeptide caspase inhibitor or a specific tetrapeptide inhibitor of caspase-3 just after SCI reduced enzyme activity at 7 days. Apoptotic cells were identified with TUNEL staining in both neurons and oligodendrocytes in mice after SCI, which also showed activated caspase-3. Increased caspase-3- and caspase-8-like activity was detected in the injured spinal cord on days 3 and 7. Caspase protease activities may be involved in delayed neuronal and glial apoptosis after SCI. PMID- 12568319 TI - Saccular aneurysm of the accessory middle cerebral artery--case report. AB - A 30-year-old man presented with a rare ruptured saccular aneurysm located at the junction of the accessory middle cerebral artery and the horizontal portion (A1 segment) of the anterior cerebral artery. A right frontotemporal craniotomy was performed and the aneurysm was clipped through a transsylvian approach. The postoperative course was uneventful and he was discharged in good condition. The transsylvian approach is the method of choice for such aneurysm. PMID- 12568320 TI - Cranial nerve pareses following wrapping of a ruptured dissecting vertebral artery aneurysm: a possible complication of cyanoacrylate glue--case report. AB - A 51-year-old female with a ruptured dissecting vertebral artery aneurysm underwent an uneventful wrapping technique using Biobond-soaked gauze through a unilateral suboccipital transcondyle approach. On the 3rd postoperative day, she developed pareses of the ipsilateral VII through XII cranial nerves. Daily intravenous administration of 300 mg of hydrocortisone was started. This treatment was continued and dosage was tapered until the 10th postoperative day. The cranial nerve pareses deteriorated until the 8th postoperative day, but slowly resolved by 3 weeks after surgery. The patient was discharged with slight hoarseness and dysphasia 5 weeks after surgery. She had only slight hoarseness at 6 months. This complication was probably due to a neural toxic response to the Biobond. PMID- 12568321 TI - Transcortical transchoroidal fissure approach for ruptured distal posterior cerebral artery (P2-P3 junction) aneurysm associated with packed intraventricular hemorrhage--two case reports. AB - A 56-year-old man with ruptured right P2-P3 junction aneurysm and a 66-year-old man with ruptured left P2-P3 junction aneurysm of the posterior cerebral artery associated with acute-stage packed intraventricular hemorrhage. The aneurysms were successfully clipped through the transcortical transchoroidal fissure approach. This approach requires less retraction of the temporal lobe, provides a wider surgical field, and the P2 segment can be easily reached. The present approach is very useful for the treatment of ruptured aneurysms at the P2-P3 junction, in particular for acute stage surgery associated with packed intraventricular hemorrhage. PMID- 12568322 TI - Ring-enhanced mass in the brain of a woman with systemic lupus erythematosus and elevated serum CA19-9 level: brain abscess or metastatic tumor?--case report. AB - A 70-year-old woman with systemic lupus erythematosus presented with a brain abscess manifesting as progressive monoparesis of the right lower extremity over 4 days. She had had no episodes of fever, and did not complain of headache or exhibit any signs of meningeal irritability. Computed tomography of the brain showed a round, low-density mass with strong ring enhancement in the left frontal lobe. Laboratory examination found a moderately elevated serum level of CA19-9, a marker of some digestive organ cancers. Together with the absence of febrile episodes, headache, and a rise in leukocyte count, the initial suspicion was metastatic brain tumor rather than brain abscess. However, diffusion-weighted magnetic resonance imaging depicted the mass as a very hyperintense area. The neuroimaging diagnosis was brain abscess. After conservative treatment with intravenous antibiotics for 6 weeks, the brain abscess completely resolved, and the patient was discharged without neurological deficits. PMID- 12568323 TI - Retroorbital hemangiopericytoma and cavernous sinus schwannoma--case report. AB - An 18-year-old male presented with severe proptosis and blindness in the right eye. Neuroimaging revealed a large and hypervascular tumor in the right retrobulbar region and a large tumor in the left cavernous sinus. Angiography showed the right tumor was extensively vascular, fed by a hypertrophic ophthalmic artery, and the left tumor was moderately vascular, fed by a large middle meningeal artery. Following embolization of the feeder vessels, the right retrobulbar hemangiopericytoma and the left cavernous sinus schwannoma were uneventfully and successfully resected. Such combinations of different pathological lesions present unusual therapeutic challenges. PMID- 12568324 TI - Suprasellar peri-infundibular ectopic prolactinoma--case report. AB - A 21-year-old man presented with bitemporal hemianopia and hyperprolactinemia. Magnetic resonance (MR) imaging showed a suprasellar cystic tumor in contact with the pituitary stalk. The diagnosis was craniopharyngioma. Intraoperatively, there was no clear continuity between the tumor and the tissue of the anterior lobe of the pituitary gland. The pituitary stalk and the diaphragma sellae were intact, and their morphology remained almost completely normal after the tumor was removed. The histological diagnosis was prolactin-producing pituitary adenoma. Postoperatively, the bitemporal hemianopia improved, and the serum prolactin levels returned to normal. The final diagnosis was suprasellar ectopic pituitary adenoma arising in the peri-infundibular region. Follow-up MR imaging at 1 year showed a normal pituitary stalk and pituitary gland, with no evidence of residual tumor. PMID- 12568325 TI - Cervical myelopathy caused by hypoplasia of the atlas and ossification of the transverse ligament--case report. AB - A 79-year-old Japanese female presented with symptomatic cervical myelopathy caused by a hypoplastic posterior arch of the atlas and ossification of the transverse ligament. Neuroradiological examination demonstrated a hypoplastic posterior arch of the atlas and ossification of the transverse ligament. The cervical spinal cord was compressed at the level of the atlas by both the hypoplastic posterior arch of the atlas and the ossification of the transverse ligament. The patient underwent C-1 laminectomy, which arrested the progressive myelopathy and resulted in a good recovery. Atlas hypoplasia with ossification of the transverse ligament may be associated with Asian ethnicity. PMID- 12568326 TI - Introduction to the multi-author review on conjugative transposons. PMID- 12568327 TI - Mechanism of integration and excision in conjugative transposons. AB - Translocation of conjugative transposons proceeds via excision of the element to generate a circular molecule that can then integrate into a new site, which can be in the same or a different cell. This review summarises some of the different mechanisms used for excision and integration of conjugative transposons. PMID- 12568328 TI - Association of different mobile elements to generate novel integrative elements. AB - Among the more important problems in modern hospitals is the prevalence of bacterial pathogens expressing resistance to multiple antimicrobial agents. The frequency of multiresistance suggests mechanisms by which bacterial species can concentrate and efficiently exchange a variety of resistance determinants. Mechanisms by which this occurs include insertion of transposons within transposons, coalescence through the activity of insertion sequences and the employment of integrons. In some instances, more than one of these mechanisms is involved in creating large multiresistance genetic elements. The association of the elements with transferable elements or transposons may promote rapid dissemination among clinical strains, and create further opportunities for inclusion of additional resistance determinants. PMID- 12568329 TI - The clostridial mobilisable transposons. AB - Mobilisable transposons are transposable genetic elements that also encode mobilisation functions but are not in themselves conjugative. They rely on coresident conjugative elements to facilitate their transfer to recipient cells. Clostridial mobilisable transposons include Tn4451 and Tn4452 from Clostridium perfringens, and Tn4453a and Tn4453b from Clostridium difficile, all of which are closely related, and Tn5398 from C. difficile. The Tn4451 group of elements encodes resistance to chloramphenicol and is unusual in that transposition is dependent upon a large resolvase protein rather than a more conventional transposase or integrase. This group of elements also encodes the mobilisation protein TnpZ that, by acting at the RS(A) or oriT site located on the transposon, and in the presence of a coresident conjugative element, promotes the movement of the nonreplicating circular intermediate and of plasmids on which the transposon resides. The erythromycin resistance element Tn5398 is unique in that it encodes no readily identifiable transposition or mobilisation proteins. However, the element is still capable of intraspecific transfer between C. difficile isolates, by an unknown mechanism. The detailed analysis of these mobilisable clostridial elements provides evidence that the evolution and dissemination of antibiotic resistance genes is a complex process that may involve the interaction of genetic elements with very different properties. PMID- 12568330 TI - The role of Bacteroides conjugative transposons in the dissemination of antibiotic resistance genes. AB - Investigations into the mechanisms of antibiotic resistance gene transfer utilized by Bacteroides species have led to a greater understanding of how bacteria transfer antibiotic resistance genes, and what environmental stimuli promote such horizontal transfer events. Although Bacteroides spp. harbor a variety of transmissible elements that are involved in the dissemination of antibiotic resistance genes, it is one particular class of elements, the conjugative transposons, that are responsible for most of the resistance gene transfer in Bacteroides. The potential for Bacteroides conjugative transposons to transfer antibiotic resistance genes extends beyond those genes carried by the conjugative transposon itself, because Bacteroides conjugative transposons are able to mobilize coresident plasmids in trans and in cis, and also stimulate the excision and transfer of unlinked integrated elements called mobilizable transposons. These characteristics of conjugative transposons alone have significant implications for the ecology and spread of antibiotic resistance genes, and in terms of biotechnology. A novel feature of the most widespread family of Bacteroides conjugative transposons, the CTnDOT/ERL family, is that their transfer is stimulated 100- to 1000-fold by low concentrations of tetracycline. This is significant because the use of antibiotics not only selects for resistant Bacteroides strains, but also stimulates their transfer. Other Bacteroides conjugative transposons do not require any induction to stimulate transfer, and hence appear to transfer constitutively. The constitutively transferring elements characterized so far appear to have a broader host range than the CTnDOT/ERL family of conjugative transposons, and the prevalence of these elements is on the increase. Since these constitutively transferring elements do not require induction by antibiotics to stimulate transfer, they have the potential to become as pervasive as the CTnDOT/ERL family of conjugative transposons. PMID- 12568331 TI - The role of conjugative transposons in the Enterobacteriaceae. AB - Although widely studied in gram-positive Streptococci and in the gram-negative Bacteroides, there is a scarcity of information on the occurrence and nature of conjugative transposon-like elements in the well-studied Enterobacteriaceae. In fact, some of the major reviews on conjugative transposons prior to 1996 failed to mention their occurrence in this group. Recently, their presence has been reported in Salmonella, Vibrio and Proteus species, and in some cases such as the SXT element in Vibrio and the IncJ group element CTnR391, there has been some molecular characterization. The elements thus far examined appear to be larger than the common gram-positive conjugative transposons and to be mosaic in structure, with genes derived from several sources. Recent evidence suggests that in the Enterobacteriaceae the elements may be related to enteric pathogenicity islands. The evolution, distribution and role of these elements in the Enterobacteriaceae is discussed. PMID- 12568332 TI - Comparison of SXT and R391, two conjugative integrating elements: definition of a genetic backbone for the mobilization of resistance determinants. AB - The SXT element (SXT) is becoming an increasingly prevalent vector for the dissemination of antibiotic resistances in Vibrio cholerae. SXT is a member of a larger family of elements, formerly defined as IncJ plasmids, that are self transmissible by conjugation and integrate site-specifically into the host chromosome. Comparison of the DNA sequences of SXT and R391, an IncJ element from Providencia rettgeri, indicate that these elements consist of a conserved backbone that mediates the regulation, excision/integration and conjugative transfer of the elements. Both elements have insertions into this backbone that either confer the element-specific properties or are of unknown function. Interestingly, the conserved SXT and R391 backbone apparently contains hotspots for insertion of additional DNA sequences. This backbone represents a scaffold for the mobilization of genetic material between a wide range of gram-negative bacteria, allowing for rapid adaptation to changing environments. PMID- 12568333 TI - The role of conjugative transposons in spreading antibiotic resistance between bacteria that inhabit the gastrointestinal tract. AB - There is huge potential for genetic exchange to occur within the dense, diverse anaerobic microbial population inhabiting the gastrointestinal tract (GIT) of humans and animals. However, the incidence of conjugative transposons (CTns) and the antibiotic resistance genes they carry has not been well studied among this population. Since any incoming bacteria, including pathogens, can access this reservoir of genes, this oversight would appear to be an important one. Recent evidence has shown that anaerobic bacteria native to the rumen or hindgut harbour both novel antibiotic resistance genes and novel conjugative transposons. These CTns, and previously characterized CTns, can be transferred to a wide range of commensal bacteria under laboratory and in vivo conditions. The main evidence that gene transfer occurs widely in vivo between GIT bacteria, and between GIT bacteria and pathogenic bacteria, is that identical resistance genes are present in diverse bacterial species from different hosts. PMID- 12568334 TI - Retrovirus molecular conjugates. AB - Retrovirus-derived vectors are currently the preferred vectors used for human gene therapy protocols. Serious safety concerns persist, however, which are specifically related to the formation of a replication-competent virus, and no synthesis method currently employed precludes its formation with certainty. For many cell types, a low transduction efficiency results in insufficient therapeutic benefit. We describe the development of a molecular conjugate system, which permits transient chemical modification of a retrovirus with polylysine. This modification not only introduces additional safety features over standard unmodified retrovirus vectors, but also provides enhanced transduction efficiency. PMID- 12568335 TI - The functions of mucosal T cells in containing the indigenous commensal flora of the intestine. AB - There is an immense load of non-pathogenic commensal bacteria in the distal small intestine and the colon of mammals. The physical barrier that prevents penetration (translocation) of these organisms into the body is a simple epithelium comprised of the single enterocyte/colonocyte cell layer with its overlying mucus. In this review, we discuss the roles of intestinal T cells in initiating and regulating innate and adaptive mucosal immune responses of the mucosal immune system that avoid or limit penetration of the commensal intestinal bacteria. PMID- 12568336 TI - Dictyostelium mobile elements: strategies to amplify in a compact genome. AB - Dictyostelium discoideum is a eukaryotic microorganism that is attractive for the study of fundamental biological phenomena such as cell-cell communication, formation of multicellularity, cell differentiation and morphogenesis. Large scale sequencing of the D. discoideum genome has provided new insights into evolutionary strategies evolved by transposable elements (TEs) to settle in compact microbial genomes and to maintain active populations over evolutionary time. The high gene density (about 1 gene/2.6 kb) of the D. discoideum genome leaves limited space for selfish molecular invaders to move and amplify without causing deleterious mutations that eradicate their host. Targeting of transfer RNA (tRNA) gene loci appears to be a generally successful strategy for TEs residing in compact genomes to insert away from coding regions. In D. discoideum, tRNA gene-targeted retrotransposition has evolved independently at least three times by both non-long terminal repeat (LTR) retrotransposons and retrovirus-like LTR retrotransposons. Unlike the nonspecifically inserting D. discoideum TEs, which have a strong tendency to insert into preexisting TE copies and form large and complex clusters near the ends of chromosomes, the tRNA gene-targeted retrotransposons have managed to occupy 75% of the tRNA gene loci spread on chromosome 2 and represent 80% of the TEs recognized on the assembled central 6.5 Mb part of chromosome 2. In this review we update the available information about D. discoideum TEs which emerges both from previous work and current large-scale genome sequencing, with special emphasis on the fact that tRNA genes are principal determinants of retrotransposon insertions into the D. discoideum genome. PMID- 12568337 TI - Protecting against promiscuity: the regulatory role of insulators. AB - Eukaryotic genomes contain transcriptional regulatory elements that alter promoter activity through long-range interactions. Many control elements show a broad range of promoter interactions, suggesting that these elements are capable of inappropriate transcription. The identification of a novel class of directing regulatory elements, called insulators, has provided clues into mechanisms used in eukaryotic genomes to maintain transcription fidelity. Insulators contribute to the organization of independent domains of gene function by restricting enhancer and silencer function. This review describes the properties of insulators and related elements that have been isolated from several eukaryotic genomes. Two classes of models of insulator function are considered. These models provide insights into possible mechanisms used by these diverse elements to provide regulatory autonomy. PMID- 12568338 TI - The subventricular zone: new molecular and cellular developments. AB - The subventricular zone (SVZ), which lines the lateral walls of the lateral ventricle, persists as a neurogenic zone into adulthood and functions as the largest site of neurogenesis in the adult brain. In recent years, with the acceptance of the concept of postembryonic mammalian neurogenesis, neurogenesis in the adult SVZ has been an area of active research. With the rapid accumulation of new information on the SVZ, some of which is contradictory, summarizing existing knowledge on the SVZ and outlining future research directions in this area become important. In this review, we will cover recent molecular and cellular investigations that characterize the SVZ niche, SVZ neurogenesis, and SVZ cell migration within the adult brain. PMID- 12568339 TI - Active-site mutants of class B beta-lactamases: substrate binding and mechanistic study. AB - Increased resistance to beta-lactam antibiotics is mainly due to beta-lactamases. X-ray structures of zinc beta-lactamases unraveled the coordination of the metal ions, but their mode of action remains unclear. Recently, enzymes in which one of the zinc ligands was mutated have been characterized and their catalytic activity against several beta-lactam antibiotics measured. A molecular modeling study of these enzymes was performed here to explain the catalytic activity of the mutants. Coordination around the zinc ions influences the way the tetrahedral intermediate is bound; any modification influences the first recognition of the substrate by the enzyme. For all the studied mutants, at least one of the interactions fails, inducing a loss of catalytic efficiency compared to the wild type. The present studies show that the enzyme cavity is a structure of high plasticity both structurally and mechanistically and that local modifications may propagate its effects far from the mutated amino acid. PMID- 12568340 TI - Structural features of prions explored by sequence analysis. II. A PrP(Sc) model. AB - Prion diseases are neurodegenerative disorders associated with a conformational conversion of the prion PrP protein, in which the beta strand content increases and that of the a helix decreases. However, the structure of the pathogenous form PrP(Sc), occurring after conformational conversion of the normal cellular form PrP(C), is not yet known. From sequence analysis, we have previously proposed that helix H2 of the prion PrP(C) structure might be a key region for this structural conversion. More recently, we identified the TATA box-binding protein fold as a putative scaffold that may locally satisfy the predicted secondary structure organisation of PrP(Sc). In the present analysis, we detail the schematic construction of PrP(Sc) monomeric and dimeric models, based on this hypothesis. These models are globally compatible with available data and therefore may provide further insights into the structurally and functionally elusive PrP protein. Some comments are also devoted to a comparison of the yeast Ure2p prion and animal prions. PMID- 12568341 TI - Specific localization of the catalytic subunits of protein kinase CK2 at the centrosomes. AB - The protein kinase CK2 holoenzyme is composed of two regulatory beta subunits and two catalytic alpha or alpha' subunits. Although experimental evidence for involvement of the enzyme in the regulation of cell proliferation is accumulating, the exact mechanism of its action is still unclear. The subcellular localization of the enzyme may be a key to its function. We have recently shown that the CK2 holoenzyme is tightly associated with the Golgi complex and the endoplasmic reticulum. Centrosomes, which organize spindle formation during the cell cycle and microtubule cytoskeleton formation and, thereby, the location and orientation of different organelles in the cell, are in close vicinity to the Golgi complex. Because several kinases and phosphatases have been described to regulate the functions of the centrosome, we analysed the association of CK2 with these organelles. Using biochemical cell fractionation and coimmunoprecipitation, we never found the holoenzyme but only the catalytic alpha subunits associated with the centrosome. These data were confirmed by immunoelectron microscopy. Thus, the present data point to a particular role of the catalytic alpha and alpha' subunit of protein kinase CK2, which may be different from their roles in the holoenzyme. PMID- 12568342 TI - ERKs are the point of divergence of PKA and PKC activation by PTHrP in human skin fibroblasts. AB - Parathyroid hormone-related peptide (PTHrP) receptors, coupled to trimeric G proteins, operate in most target cells through at least three different transduction routes: Galpha s-mediated stimulation of adenylylcyclase (AC), Galpha q-mediated activation of phospholipase Cbeta (PLC) and mitogen-activated protein kinase (MAPK) activation. In this study we investigated the relative role of different pathways in human skin fibroblast proliferation. Using chemical inhibitors and activators of signal transduction, we demonstrated that: (i) AC/cAMP and PLC/1,4,5 inositol triphosphate/diacylglycerol second-messenger systems are simultaneously activated following PTHrP binding to its receptors; (ii) the mitogenic response to PTHrP derives from a balance between two counteracting pathways--an activating route mediated by protein kinase C (PKC) and an inhibitory route mediated by protein kinase A (PKA); (iii) PTHrP mitogenic effects are largely dependent on MAPKs, whose activity can be modulated by both PKA and PKC. Our results indicate that MAPKs are common targets of both transduction routes and, at the same time, their point of divergence in mediating PTHrP dual and opposite mitogenic effects. PMID- 12568343 TI - Agonist-induced internalization and desensitization of the human nociceptin receptor expressed in CHO cells. AB - In this study, we examined agonist-induced internalization, recycling and signalling (measure of cAMP levels) of the cloned human nociceptin receptor (hNOP) expressed in CHO-K1 cells. Internalization was proven by a receptor binding assay on viable cells. The agonist nociceptin/orphanin FQ (NC) promoted rapid internalization of the hNOP receptor (approximately 78% of cell surface receptors were lost after 2 min exposure to 1 microM NC) in a clathrin- and ATP dependent manner. Internalization was more rapid and marked in CHO-K1 cells than, as we previously reported, in SK-N-BE cells. This difference may be related to higher levels of beta-arrestin isoforms detected in CHO-K1 than in SK-N-BE cells. hNOP receptor internalization was partially reversible and recycling occurred in the presence of the agonist; receptor recycling was dependent on okadaic acid sensitive phosphatases and was blocked by monensin. Confocal microscopy analysis confirmed the internalization and the recycling back to the plasma membrane of an epitope-tagged hNOP receptor expressed in CHO-K1 cells. These receptors underwent rapid desensitization upon agonist challenge: NC efficacy in inhibiting forskolin stimulated cAMP production was significantly reduced 10 min after exposure and correlated with the rate of receptor internalization. Moreover, we observed that blockade of hNOP receptor recycling by monensin would cause a more prolonged and relevant desensitization of this receptor. Thus, the dynamic cycle between hNOP receptor activation, internalization and recycling determines the activity of this receptor on the cell surface. PMID- 12568344 TI - Susceptibility of recombinant porcine endogenous retrovirus reverse transcriptase to nucleoside and non-nucleoside inhibitors. AB - Transplantation of organs, tissues or cells from pigs to humans could be a potential solution to the shortage of human organs for transplantation. Porcine endogenous retroviruses (PERVs) remain a major safety concern for porcine xenotransplantation. Thus, finding drugs that could be used as virological prophylaxis (or therapy) against PERV replication would be desirable. One of the most effective ways to block retroviral multiplication is to inhibit the enzyme reverse transcriptase (RT) which catalyzes the reverse transcription of viral RNA to proviral double-stranded DNA. We report here the cloning and expression of PERV RT and its susceptibility to several inhibitors. Our data demonstrate PERV susceptibility in vitro to the triphosphorylated nucleoside analog of zidovudine (AZT) and to ddGTP and to a lesser extent to ddTTP but almost no susceptibility to the non-nucleoside RT inhibitors tested. PMID- 12568345 TI - The processing and presentation of endogenous and exogenous antigen by Schwann cells in vitro. AB - The expression of major histocomatibility complex class II in vitro and in vivo by Schwann cells indicates a potential facultative role of Schwann cells in the presentation of antigen to neuritogenic T cells during inflammatory demyelinating neuropathies. Using a T cell proliferation assay, this study demonstrated that processing and presentation of endogenous and exogenous antigen by Schwann cells influences T cell proliferation. Statistical analysis of proliferation and its relation to processing and presentation of antigen by Schwann cells had not been previously addressed. Different combinations of factors including treatment of cultures (untreated, irradiated or fixed), concentration of exogenous antigen (0 or 40 microg/ml), the presence of interferon-gamma and the timing of exogenous antigen addition influence the proliferation P2-specific, non-mammalian protein ovalbumin-specific T cell lines and naive T cells. PMID- 12568346 TI - Morphofunctional changes in the mitochondrial subpopulations of conceptus tissues during the placentation process. AB - To establish the role of mitochondrial subpopulations in the mitochondrial maturation process, we studied morphological and functional changes in the mitochondria of different mammalian conceptus tissues during the organogenic and the placentation processes. Mitochondrial subpopulations of three different conceptus tissues, embryo and visceral yolk sac placenta on gestational days 11, 12 and 13 and placenta on days 12 and 13, were examined morphologically by transmission electron microscopy. Cytochrome oxidase activity and protein levels were also measured in each mitochondrial subpopulation. The results indicate two different mitochondrial subpopulation profiles: a homogeneous one, which corresponds to immature mitochondria, and a heterogeneous one, which represents the mature mitochondria. The three tissues studied show different morphologic and metabolic patterns of mitochondrial maturation during the placentation process, rendering them suitable as experimental models to establish the possible relationship between mitochondrial maturation and the mitochondrial subpopulations. PMID- 12568347 TI - Isolation of an antifreeze peptide from the Antarctic sponge Homaxinella balfourensis. AB - Polar plants and animals survive in subzero waters (-2 degrees C) and many of these marine organisms produce antifreeze proteins (AFPs) to better adapt themselves to these conditions. AFPs prevent the growth of ice crystals which disrupt cellular membranes and destroy cells by inhibiting crystallization of water within the organism. The hydrophilic extract of an Antarctic sponge Homaxinella balfourensis exhibited a non-colligative freezing point depression effect on the crystal morphology of water. The extract was purified by repeated reverse phase high-pressure liquid chromatography, then assayed and shown to contain several AFPs. The major peptide was isolated, analyzed using matrix assisted laser desorption ionization mass spectrometry and the partial structure of the peptide identified through amino acid sequencing. AFPs have potential applications in agriculture, medicine and the food industry. PMID- 12568348 TI - Isoforms of soluble alpha-tubulin in oocytes and brain of the frog (genus Rana): changes during oocyte maturation. AB - Rana oocytes have previously been shown to contain much more soluble tubulin than does the brain, suggesting different assembly and disassembly dynamics of frog oocyte tubulin compared to that in brain. By using centrifugation, SDS-PAGE, two dimensional gel electrophoresis and Western blots, probed with anti-alpha-tubulin monoclonal antibodies, polymorphic alpha-tubulins (isoforms) were compared in brains and follicle-enclosed oocytes of northern (Rana pipiens) and southern (R. berlandieri) frogs. Oocyte tubulin in both species had isoforms with greater ranges of isoelectric point (pI) than those of brain tubulins; in particular, the oocyte tubulin pIs ranged further into the acidic region of the isoelectric focusing gels than corresponding brain tubulin. This difference may, in part, be responsible for the previously reported assembly differences between oocyte tubulin (undetectable assembly) and brain tubulin (high assembly). Isoforms of alpha-tubulin with relatively acidic pI were more abundant in northern frog brain and oocyte soluble extracts than in analogous extracts from southern frogs. Furthermore, additional acidic alpha-tubulin isoforms were found in progesterone treated oocytes (i.e., eggs), indicating increased heterogeneity of acidic alpha tubulin isoforms during oocyte meiotic maturation. Among northern frog oocyte soluble components fractionated on Superose-6b columns, tubulin complexes with apparent molecular mass of about 1800 kDa were found to contain acidic alpha tubulin isoforms while the putative oligomeric tubulins with an apparent molecular mass of about 250 kDa contained an additional relatively basic alpha tubulin isoform. The acidic alpha-tubulin isoforms, therefore, are proposed to be associated with cold-adaptable cells of brain and oocytes, and may also be involved in stabilization of large soluble tubulin complexes in oocytes of the northern frog. PMID- 12568349 TI - Synthesis and antiinflammatory activity of novel indazolones. AB - In this study, a series of new N2 substituted 1,2-dihydro-3H-indazol-3-ones (3a d) as well as their condensed pyrazolo, pyridazino derivatives such as pyridazino[1,2-a]indazole-6,9,11-triones (4a-h) and 3,9-dioxo-3H,9H-pyrazolo[1,2 a]indazole (7) were synthesized. The antiinflammatory activity of some synthesized compounds was determined by carrageenan-induced rat paw edema technique using diclofenac as reference drug. The pharmacological data showed that most of the tested compounds exhibited a significant long lasting antiinflammatory activity, which in the case of compound 3b was superior to that of diclofenac. PMID- 12568350 TI - Importance of sulfonylimidazolidinone motif of 4-phenyl-1 arylsulfonylimidazolidinones for their cytotoxicity: synthesis of 2-benzoyl-4 phenyl[1,2,5]thiazolidine-1,1-dioxides and their cytotoxcity. AB - For probing the importance of planarity of imidazolidinone motif of 4-phenyl-1 (benzenesulfonyl)imidazolidinones 1 for their cytotoxicity, 4-phenyl-2 (benzoyl)[1,2,5]thiadiazolidine-1,1-dioxide (2a), 4-phenyl-2-(p toluoyl)[1,2,5]thiadiazolidine-1,1-dioxide (2b), 4-phenyl-2 (phenylcarbamoyl)[1,2,5]thiadiazolidine-1,1-dioxide (3a), and 4-phenyl-2-(p tolylcarbamoyl)[1,2,5]thiadiazolidine-1,1-dioxide (3b) were prepared along with their regioisomers (5a, 5b, 9a, 9b) and their cytotoxicity were measured against human lung carcinoma (A549), human colon carcinoma (COL0205), human ovarian cancer (SK-OV-3), human leukemic cancer (K562), and murine colon adenocarcinoma (Colon26) cell lines in vitro. All compounds prepared do not show any activity against all five cancer cell lines unlike 1. Compounds 1 possess planarity of imidazolidinone, especially in sulfonylurea moiety (-SO2NHCONH-). However compounds 2 and 3 have nonplanar 5-membered ring, [1,2,5]thiadiazolidine-1,1 dioxides. Such structural differentiation might result in the loss of activity. Therefore the inactivity of 2 and 3 could also be an indication for the necessity of planarity of imidazolidinone ring of 1 for their cytotoxic activity. PMID- 12568351 TI - Chemical constituents from the Hydrangea chinensis. AB - Two quinazolone alkaloids, (+)-febrifugine (1) and isofebrifugine (2), along with three coumarin derivatives, 6-hydroxy coumarin (3), skimmin (5), and umbelliferone-7-O-alpha-L-rhamnopyranosyl(1-->4)-beta-D-glucopyranoside (6), were isolated from the roots of Hydrangea chinensis. Compound 6 is a new compound. In addition, umbelliferone (4), linoleic acid (7), two steroidal glycosides (8, 9), three furfural derivatives (10-12), and butyl-beta-D-fructofuranoside (13) were isolated from the leaves of the same plant. The structures of all isolates were elucidated by spectral methods. PMID- 12568352 TI - Indolyl alkaloid derivatives, Nb-acetyltryptamine and oxaline from a marine derived fungus. AB - Indolyl alkaloids, Nb-acetyltryptamine (1) and the known oxaline (2) have been isolated from the organic extract of the broth of an unidentified fungus collected from the surface of the marine red alga Gracilaria verrucosa. The structure of Nb-acetyltryptamine (1) was assigned on the basis of comprehensive spectroscopic analyses. PMID- 12568353 TI - Further isolation of antioxidative (+)-1-hydroxypinoresinol-1-O-beta-D-glucoside from the rhizome of Salvia miltiorrhiza that acts on peroxynitrite, total ROS and 1,1-diphenyl-2-picrylhydrazyl radical. AB - A furanofuranoid lignan glycoside, with radical scavenging on peroxynitrite, total reactive oxygen species (ROS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, was isolated from the rhizome of Salvia miltiorrhiza and characterized as (+)-1-hydroxypinoresinol-1-O-beta-D-glucoside based on spectroscopic evidence. The compound exhibited peroxynitrite, total ROS and DPPH radical scavenging activities with IC50 values of 3.23 +/- 0.04, 2.26 +/- 0.07 and 32.3 +/- 0.13 microM, respectively. Penicillamine, Trolox (6-hydroxy-2,5,7,8-tetramethylchroman 2-carboxylic acid) and L-ascorbic acid, acting as positive controls, showed radical scavenging activities with IC50 values of 6.72 +/- 0.25, 1.43 +/- 0.04 and 11.4 +/- 0.07 microM, respectively. PMID- 12568354 TI - Effects of ginsenosides on GABA(A) receptor channels expressed in Xenopus oocytes. AB - Ginsenosides, major active ingredients of Panax ginseng, are known to regulate excitatory ligand-gated ion channel activity such as nicotinic acetylcholine and NMDA receptor channel activity. However, it is not known whether ginsenosides affect inhibitory ligand-gated ion channel activity. We investigated the effect of ginsenosides on human recombinant GABA(A) receptor (alpha1beta1gamma2S) channel activity expressed in Xenopus oocytes using a two-electrode voltage-clamp technique. Among the eight individual ginsenosides examined, namely, Rb1, Rb2, Rc, Rd, Re, Rf, Rg1 and Rg2, we found that Rc most potently enhanced the GABA induced inward peak current (I(GABA)). Ginsenoside Rc alone induced an inward membrane current in certain batches of oocytes expressing the GABA(A) receptor. The effect of ginsenoside Rc on I(GABA) was both dose-dependent and reversible. The half-stimulatory concentration (EC50) of ginsenoside Rc was 53.2 +/- 12.3 microM. Both bicuculline, a GABA(A) receptor antagonist, and picrotoxin, a GABA(A) channel blocker, blocked the stimulatory effect of ginsenoside Rc on I(GABA). Niflumic acid (NFA) and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), both Cl- channel blockers, attenuated the effect of ginsenoside Rc on I(GABA). This study suggests that ginsenosides regulated GABA(A) receptor expressed in Xenopus oocytes and implies that this regulation might be one of the pharmacological actions of Panax ginseng. PMID- 12568355 TI - Induction of growth hormone by the roots of Astragalus membranaceus in pituitary cell culture. AB - The traditional Asian medicinal herb, roots of Astragalus (A.) membranaceus (Leguminosae), is used for many purposes, some of which are purported to stimulate the release of growth hormone in vivo. Extracts of A. membranaceus were tested to determine whether they stimulate the release of growth hormone in rat pituitary cell culture. A. membranaceus was extracted sequentially with 80% ethanol (fraction A), n-hexane (fraction B); the test compound from the herbal extraction was isolated using silica gel column chromatography and was identified with spectral data. Test compound was also extracted by traditional boiling water methods. Induction of growth hormone in pituitary cell culture was conducted with isolated compounds and extracted fractions of A. Radix (dried roots of A. membranaceus). The fraction A was not active in the rat pituitary cell culture, but the fraction B derived from the ethanol fraction stimulated the release of growth hormone in culture. Six compounds from fraction B (1-6) were isolated and identified previously. The compounds 1,2-benzendicarboxylic acid diisononylester (1), beta-sitosterol (2), and 3-O-beta-D-galactopyranosyl-beta-sitosterol (5) did not induce growth hormone release in the culture. Formononetin (3), 9Z,12Z octadecadienoic acid (4), stigmast-4-en-6beta-ol-3-one (6) and 98-E, a mixture of 1'-9,12-octadecadienoic acid (Z,Z)-2',3'-dihydroxy-propylester (7) and 1' hexadecanoic acid-2',3'-dihydroxy-propylester (8) stimulated the release of growth hormone in the rat pituitary cell culture significantly compared to the control. In conclusions, four compounds isolated from extracts of A. Radix induced growth hormone release in the rat pituitary cell culture. The 98-E isolate was the most active inducer of growth hormone release. PMID- 12568356 TI - Antibacterial compounds from the leaves of Acanthopanax senticosus. AB - Chiisanogenin (1), hyperin (2) and chiisanoside (3) were isolated from the leaves of Acanthopanax senticosus, and were tested for their inhibitory activities against 6 strains of bacteria. Among them, chiisanogenin (1) revealed broad but moderate antibacterial activities against G (+) and G (-) bacteria, the minimum inhibitory concentration (MIC) being in the range of 50-100 microg/ml. PMID- 12568357 TI - In vivo anti-oxidant activities of tectochrysin. AB - The anti-oxidant activities of tectochrysin, a major compound of propolis, were investigated. Tectochrysin exhibited a significant decrease in serum transaminase activities elevated by hepatic damage induced by CCl4-intoxication in rats. Tectochrysin tested exhibited a lipid peroxidation causing a significant decrease in MDA production in TBA-reactant assay. Tectochrysin was strong in the increase in the anti-oxidant enzymes such as hepatic cytosolic superoxide dismutase, catalase and glutathione peroxidase activities in CCl4-intoxicated rats. These results suggest that tectochrysin possess not only the anti-oxidant, but also the activities in CCl4-intoxicated rats. Especially, tectochrysin was found to cause significant increases in the rat liver cytosolic SOD, catalase, GSH-px activities as well as a significant decrease in the MDA production. PMID- 12568358 TI - The roles of Kupffer cells in hepatocellular dysfunction after femur fracture trauma in rats. AB - The aim of this study was to investigate the effects of trauma on alterations in cytochrome P450 (CYP 450)-dependent drug metabolizing function and to determine the role of Kupffer cells in hepatocellular dysfunction. Rats underwent closed femur fracture (FFx) with associated soft-tissue injury under anesthesia, while control animals received only anesthesia. To deplete Kupffer cells in vivo, gadolinium chloride (GdCl3) was injected intravenously via the tail vein at 7.5 mg/kg body wt., 1 and 2 days prior to FFx surgery. At 72 h after FFx, serum alanine aminotransferase (ALT) activity was increased, and this increase was attenuated by GdCl3 pretreatment. Serum aspartate aminotransferase (AST) and lipid peroxidation levels were not changed by FFx. Hepatic microsomal CYP 450 content and aniline p-hydroxylase (CYP 2E1) activity were significantly decreased; decreases that were not prevented by GdCl3. The level of CYP 2B1 activity was decreased by Kupffer cell inactivation, but not by FFx. There were no significant differences in the activities of CYP 1A1, CYP 1A2 and NADPH-CYP 450 reductase among any of the experimental groups. Our findings suggest that FFx trauma causes mild alterations of hepatic CYP 450-dependent drug metabolism, and that Kupffer cells are not essential for the initiation of such injury. PMID- 12568359 TI - Ginsenoside Rc and Re stimulate c-fos expression in MCF-7 human breast carcinoma cells. AB - We have found that ginsenoside Rc and Re induce c-fos in MCF-7 human breast carcinoma cells at both the mRNA and protein levels. However, neither ginsenoside activated the expression of reporter gene under the control of AP-1/TPA response elements. We have also examined the possibility that ginsenoside Rc and Re act by binding to intracellular steroid hormone receptors that act as transcriptional factors in the nucleus in inducing c-fos mRNA in MCF7 human breast carcinoma cells. However, ginsenoside Rc and Re did not bind to glucocorticoid, androgen, estrogen, or retinoic acid receptors as examined by the transcription activation of the luciferase reporter genes in CV-1 cells that were transiently transfected with the corresponding steroid hormone receptors and hormone responsive luciferase reporter plasmids. These data demonstrate that ginsenoside Rc and Re act via other transcription factors and not via estrogen receptor in c-Fos expression. PMID- 12568360 TI - Ginsenoside-Rb1 acts as a weak phytoestrogen in MCF-7 human breast cancer cells. AB - Ginseng has been recommended to alleviate the menopausal symptoms, which indicates that components of ginseng very likely contain estrogenic activity. We have examined the possibility that a component of Panax ginseng, ginsenoside-Rb1, acts by binding to estrogen receptor. We have investigated the estrogenic activity of ginsenoside-Rb1 in a transient transfection system using estrogen responsive luciferase plasmids in MCF-7 cells. Ginsenoside-Rb1 activated the transcription of the estrogen-responsive luciferase reporter gene in MCF-7 breast cancer cells at a concentration of 50 microM. Activation was inhibited by the specific estrogen receptor antagonist ICI 182,780, indicating that the estrogenic effect of ginsenoside-Rb1 is estrogen receptor dependent. Next, we evaluated the ability of ginsenoside-Rb1 to induce the estrogen-responsive gene c-fos by semi quantitative RT-PCR assays and Western analyses. Ginsenoside-Rb1 increased c-fos both at mRNA and protein levels. However, ginsenoside-Rb1 failed to activate the glucocorticoid receptor, the retinoic acid receptor, or the androgen receptor in CV-1 cells transiently transfected with the corresponding steroid hormone receptors and hormone responsive reporter plasmids. These data support our hypothesis that ginsenoside-Rb1 acts a weak phytoestrogen, presumably by binding and activating the estrogen receptor. PMID- 12568361 TI - Analysis of vasopressin-induced Ca2+ increase in rat hepatocytes. AB - To analyze vasopressin-induced Ca2+ increase in liver cells, rat hepatocytes were isolated and attached to collagen-coated cover slips. Using fura-2, a Ca2+ sensing dye, changes in intracellular Ca2+ concentration by vasopressin were monitored. Results in this communication suggested that vasopressin-induced Ca2+ increase were composed of both Ca2+ release from internal Ca2+ stores and influx from the plasma membrane. The Ca2+ influx consisted of two distinguishable components. One was dependent on the presence of vasopressin and the other was not. SK&F96365 blocked vasopressin-induced Ca2+ influx in a dose-dependent manner. Vasopressin-induced Ca2+ release from internal stores diminished in a primary culture of hepatocytes according to the culture time. However, changes in vasopressin-induced Ca2+ influx across the plasma membrane differed from changes in the Ca2+ release from internal stores, suggesting two separate signalings from receptor activation to internal stores and to the plasma membrane. PMID- 12568363 TI - Growth factor releasing porous poly (epsilon-caprolactone)-chitosan matrices for enhanced bone regenerative therapy. AB - Drug releasing porous poly(epsilon-caprolactone) (PCL)-chitosan matrices were fabricated for bone regenerative therapy. Porous matrices made of biodegradable polymers have been playing a crucial role as bone substitutes and as tissue engineered scaffolds in bone regenerative therapy. The matrices provided mechanical support for the developing tissue and enhanced tissue formation by releasing active agent in controlled manner. Chitosan was employed to enhance hydrophilicity and biocompatibility of the PCL matrices. PDGF-BB was incorporated into PCL-chitosan matrices to induce enhanced bone regeneration efficacy. PCL chitosan matrices retained a porous structure with a 100-200 microm pore diameter that was suitable for cellular migration and osteoid ingrowth. NaHCO3 as a porogen was incorporated 5% ratio to polymer weight to form highly porous scaffolds. PDGF-BB was released from PCL-chitosan matrices maintaining therapeutic concentration for 4 week. High osteoblasts attachment level and proliferation was observed from PCL-chitosan matrices. Scanning electron microscopic examination indicated that cultured osteoblasts showed round form and spread pseudopods after 1 day and showed broad cytoplasmic extension after 14 days. PCL-chitosan matrices promoted bone regeneration and PDGF-BB loaded matrices obtained enhanced bone formation in rat calvarial defect. These results suggested that the PDGF-BB releasing PCL-chitosan porous matrices may be potentially used as tissue engineering scaffolds or bone substitutes with high bone regenerative efficacy. PMID- 12568362 TI - Extensive hepatic uptake of Pz-peptide, a hydrophilic proline-containing pentapeptide, into isolated hepatocytes compared with colonocytes and Caco-2 cells. AB - The objective of the present study was to investigate the uptake process of 4 Phenylazobenzoxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (Pz-peptide), a hydrophilic and collagenase-labile pentapeptide, by isolated hepatocytes. For comparison, the uptake of Pz-peptide by Caco-2 cells and colonic cells, two known paracellular mutes of Pz-peptide, was also evaluated. A simple and sensitive reversed-phase HPLC assay method using UV detection has been developed. The coefficient of variation for all the criteria of validation were less than 15%. The method was, therefore, considered to be sutable for measuring the concentration of Pz-peptide in the biological cells. Pz-peptide was extensively uptaked into hepatocytes. The initial velocity of Pz-peptide uptake assessed from the initial slope of the curve was plotted as Eadie-Hofstee plots. The maximum velocity (Vmax) and the Michaelis constant (Km) were 0.190+/-0.020 nmol/min/10(6) cells and 12.1+/-3.23 microM, respectively. The permeability-surface area product (PS(influx)) was calculated to be 0.0157 ml/min/10(6) cells. Vmax and Km values for Caco-2 cells were calculated to be 6.22+/-0.930 pmol/min/10(6) cells and 82.8+/-8.37 microM, respectively, being comparable with those of colonocytes (6.04+/-1.03 pmol/min/10(6) cells and 87.8+/-13.2 microM, respectively). PS(influx) values for Caco-2 cells and colonocytes were calculated to be 0.0751 microl/min/10(6) cells and 0.0688 microl/min/10(6) cells, respectively. The more pronounced uptake of Pz peptide by hepatocytes, when compared with Caco-2 cells and colonocytes, is probably due to its specific transporter. In conclusion, Pz-peptide, a paracellularly transported pentapeptide in the intestine and ocular epithelia, was uptaked into hepatocytes extensively. Although Pz-peptide is able to be uptaked into the Caco-2 cells and colonocytes, it is less pronounced when compared with hepatocytes. PS(influx) values of Caco-2 cells and colonocytes for unbound Pz-peptide under linear conditions were less than 0.4% when compared with that of hepatocytes. PMID- 12568364 TI - Pharmacokinetics and bioavailability of oral cephalosporins, KR-984055 and its prodrugs, KR-999001 and KR-999002, in the rat. AB - KR-984055 is a new oral cephalosporin antibiotic with activity against both gram positive and gram-negative bacteria. Lipophilic ester-type prodrugs of KR-984055, i.e., KR-999001 and KR-999002, have been synthesized in an attempt to increase the oral bioavailability of this broad-spectrum antibiotic agent. In this study we determined the oral bioavailability of KR-984055 and its prodrugs in the rat, and evaluated the pharmacokinetic model that best describes the plasma concentration behavior following single intravenous (i.v.) and oral single dose. In addition, concentrations in plasma as well as biliary and urinary recovery of KR-984055 were determined. Also, protein binding of KR-984055 in plasma was examined in vitro. The degree of protein binding of KR-984055 was in the range of 92.09-94.77%. KR-984055 exhibited poor oral bioavailability (7.02 +/- 1.58%). The observed oral bioavailabilities of KR-984055 from KR-999001 and KR-999002 were 38.77 +/- 2.81% and 39.81 +/- 5.25%, respectively. These data were calculated from the levels of free KR-984055 in plasma. Oral KR-999001 and KR-999002 were not recovered from plasma, suggesting that it was readily cleaved to free KR 984055. KR-999001 and KR-999002 appear to be an efficient oral prodrug of KR 984055 that deserved further clinical evaluation in human. PMID- 12568365 TI - Pharmacokinetics and metabolism of endothelin receptor antagonist: contribution of kidneys in the overall in vivo N-demethylation. AB - In vivo clearance of BMS-182874 was primarily due to metabolism via stepwise N demethylation. Despite in vivo clearance approached ca 50% of the total liver plasma flow, BMS-182874 was completely bioavailable after oral administration in rats. Saturable first-pass metabolism and the role of extrahepatic tissue were evaluated as possible reasons for complete oral bioavailability despite extensive metabolic clearance. Pharmacokinetic parameters were obtained after an intravenous and a range of oral doses of BMS-182874 in rats. Bile and urine were collected from bile-duct cannulated (BDC) rats and the in vivo metabolic pathways of BMS-182874 were evaluated. Pharmacokinetics of BMS-182874 were also compared in nephrectomized (renally impaired) vs. sham-operated control rats. Oral bioavailability of BMS-182874 averaged 100%, indicating that BMS-182874 was completely absorbed and the first-pass metabolism (liver or intestine) was negligible. The AUC and Cmax values increased dose-proportionally, indicating kinetics were linear within the oral dose range of 13 to 290 mmole/kg. After intravenous administration of BMS-182874 to BDC rats, about 2% of intact BMS 182874 was recovered in excreta, indicating that BMS-182874 was cleared primarily via metabolism in vivo. The major metabolite circulating in plasma was the mono-N desmethyl metabolite and the major metabolite recovered in excreta was the di-N desmethyl metabolite. In vivo clearance of BMS-182874 was significantly reduced in nephrectomized rats. These observations suggest saturable first-pass metabolism is unlikely to be a mechanism for complete oral bioavailability of BMS 182874. Reduced clearance observed in the nephrectomized rats suggests that extrahepatic tissues (e.g., kidneys) may play an important role in the in vivo clearance of xenobiotics that are metabolized via N-demethylation. PMID- 12568366 TI - Separation methods in the chemistry of humic substances. AB - Separation methods are widely used to isolate humic substances (HSs), to fractionate them before further investigation, and to obtain information about their structure and properties. Among the chromatographic methods, techniques based on a size-exclusion effect appear to be most useful, as they allow us to relate elution data to the molecular mass distribution of HSs. The limitations of this approach are discussed in this review. Gas chromatography with mass spectrometric detection is typically used to identify the products of pyrolysis or thermochemolysis of HSs; this technique is considered most important in the structural investigation of HSs. Electrophoretic methods (especially capillary zone electrophoresis) provide detailed characterization of HSs, but it is very difficult to relate the electrophoretic data to any specific subfraction, structure or properties of HSs. The electrophoretic patterns are often called "fingerprints" and can potentially be used for the identification and classification of HSs. This is limited, however, by the great diversity of the procedures employed and by the low degree of harmonization--no data on reproducibility and between-laboratory comparability are available. The same holds true, to a certain degree, for most methods utilized for the characterization of HSs. Separation methods play an important role in the examination of the interactions of HSs with heavy metals and other chemical pollutants. They allow us to determine binding constants and other data necessary to predict the mobility of chemical pollutants in the environment. PMID- 12568367 TI - Analysis of virgin olive oil volatile compounds by headspace solid-phase microextraction coupled to gas chromatography with mass spectrometric and flame ionization detection. AB - The efficiency of headspace solid-phase microextraction (SPME) was evaluated for the qualitative and semi-quantitative analysis of virgin olive oil volatile compounds. The behaviour of four fibre coatings was compared for sensitivity, repeatability and linearity of response. A divinylbenzene-Carboxen polydimethylsiloxane fibre coating was found to be the most suitable for the analysis of virgin olive oil volatiles. Sampling and chromatographic conditions were examined and the SPME method, coupled to GC with MS and flame ionization detection, was applied to virgin olive oil samples. More than 100 compounds were isolated and characterised. The presence of some of these compounds in virgin olive oil has not previously been reported. The main volatile compounds present in the oil samples were determined quantitatively. PMID- 12568368 TI - Determination of primary aromatic amines in water food simulant using solid-phase analytical derivatization followed by gas chromatography coupled with mass spectrometry. AB - Solid phase analytical derivatization with trifluoroacetic anhydride has been introduced as sample preparation for the determination of primary aromatic amines in water by gas chromatography coupled with mass spectrometry. Water was used as a food simulant for testing migration from laminated flexible food packaging materials. The method was evaluated for 8 primary aromatic amines in 200 ml water samples, which resulted in detection limits in the 0.1-0.4 microg/l range, relative standard deviations in the 4-17% range and acceptable linearity (R2 = 0.997-1.000). Detectable levels of 2,4-diaminotoluene, 2,6-diaminotoluene and 4,4'-methylenedianiline were found in water food simulant from some of the investigated food packaging materials. PMID- 12568369 TI - Determination of rotenone in river water utilizing packed capillary column switching liquid chromatography with UV and time-of-flight mass spectrometric detection. AB - Fast and sensitive packed capillary column switching liquid chromatography methodology has been developed for the determination of the pesticide rotenone in river water. Sample volumes of up to 1 ml are loaded onto a 23 x 0.25 mm, 5 microm Kromasil C18 packed capillary precolumn using a noneluting solvent composition of water-acetonitrile (99:1, v/v) at flow-rates up to 100 microl/min prior to solute backflushing onto a 200 x 0.32 mm, 3.5 microm Kromasil C18 packed capillary analytical column using a mobile phase of water-acetonitrile (30:70, v/v) at a flow-rate of 5 microl/min. The method was evaluated using river water samples spiked with rotenone in the concentration range 0.5-50 ng/ml using UV detection. The within-assay precision was between 5.0 and 7.7% relative standard deviation (RSD, n = 6) and the between assay precision was between 7.5 and 8.9% RSD (n = 6). The method was linear within the investigated mass range displaying a calibration curve correlation factor of 0.997. The mass limit of detection was 10 pg corresponding to a concentration limit of detection of 10 pg/ml, using time of-flight mass spectrometry. PMID- 12568370 TI - Study of the mass transfer kinetics in a monolithic column. AB - The purpose of this work is to investigate the mass transfer kinetics of butylbenzoate on a monolithic RPLC column, with methanol-water (65:35, v/v) as the mobile phase. We used the perturbation method, measuring the height equivalent to a theoretical plate (HETP) of the peaks obtained as the response to small pulses of solute injected on a concentration plateau. The equilibrium isotherm of butylbenzoate was previously determined by frontal analysis. It is well accounted for by a liquid-solid extended multilayer BET isotherm model. The equilibrium data derived from the pulse method are in excellent agreement with those of frontal analysis in the accessible concentration range of 0 to 8 g/dm3. Plots of the HETP of small pulses. injected on eight different plateau concentrations, were acquired in a wide range of mobile phase flow velocities. The axial dispersion and the mass transfer kinetic coefficients were derived from these data. The validity of these measurements is discussed. The mass kinetics of butylbenzoate depends strongly on the plateau concentration. Processes involving adsorptive interactions between the solute and the stationary phase, e.g. surface diffusion and adsorption-desorption kinetics, combine in series to the external mass transfer kinetics and to effective pore diffusivity. PMID- 12568371 TI - Determination of minor conformational changes of a doxorubicin-peptide conjugate under chromatographic conditions. AB - Thermodynamic analysis of the reversed-phase retention behavior of a doxorubicin peptide conjugate demonstrated that the degree of non-linearity observed in Van't Hoff plots was impacted by mobile phase acetonitrile content over the 25-38% acetonitrile (v/v) range tested. Small decreases in the non-polar surface area of the doxorubicin-peptide conjugate as a function of temperature were estimated from these data using linear solvent strength relationships, suggesting that the retention behavior may be the result of minor analyte conformational changes during the chromatographic experiment. This hypothesis was supported via circular dichroism (CD), Raman and 1H NMR spectroscopic studies of the doxorubicin-peptide conjugate in selected chromatographic mobile phase compositions. The CD and Raman data indicated small changes to the apparent analyte microenvironment as a function of temperature and bulk solvent environment, while 1H NMR studies specifically demonstrated the environmental sensitivity of protons on three non polar peptide residues and the proximal aromatic region of the analyte. Together, these data suggest that minor changes to the conformational order of the essentially random structure of the doxorubicin-peptide conjugate are sufficient to impact chromatographic performance. PMID- 12568372 TI - Elevated-temperature ultrahigh-pressure liquid chromatography using very small polybutadiene-coated nonporous zirconia particles. AB - Capillary columns packed with small diameter particles typically lead to low permeability and long separation times in high-performance liquid chromatography. Ultrahigh pressures (>10,000 p.s.i.; 1 p.s.i. is identical with 6,894.76 Pa) can be used to overcome the limitations that small particles impose. Ultrahigh pressure liquid chromatography (UHPLC) has demonstrated great potential for high speed and high-efficiency separations. Decreasing the viscosity of the mobile phase by elevating the temperature could additionally reduce the pressure drop and facilitate the use of longer columns or smaller particles to achieve even higher total plate numbers. For this reason, we investigated the use of elevated temperatures in UHPLC. Water-resistant, flexible heater tape covered with insulation was used to provide the desired heat to the column. Polybutadiene coated 1 microm nonporous zirconia particles were used because of their chemical stability at elevated temperature. A column efficiency as high as 420,000 plates m(-1) was obtained. The effects of temperature and pressure on the separation of parabens were investigated. Separation of five herbicides was completed in 60 s using 26,000 p.s.i. and 90 degrees C. PMID- 12568373 TI - Enantioselective ion-exclusion chromatography on teicoplanin aglycone and (+)-(18 crown-6)-2,3,11,12-tetracarboxylic acid stationary phases. AB - Ion-exclusion chromatography is a well established technique for the analysis of achiral ionic species, but it has rarely been applied to chiral analytes. In this paper enantioselective ion-exclusion separations were developed on two commercially available HPLC phases: Chirobiotic TAG, based on teicoplanin aglycone, and Opticrown RCA (+), based on (+)-(18-crown-6)-2,3,11,12 tetracarboxylic acid. Chirobiotic TAG columns have a carboxylic acid group on the chiral ligand, which can be partially ionized to exclude anionic analytes by ionic repulsion. Under acidic conditions Opticrown columns have a cationic sublayer generated from the aminopropyl base silica that excludes cationic analytes. Both columns demonstrate a large dependence of efficiency on flow-rate, with the highest efficiencies at 0.1 ml/min on a 4.6 mm inner diameter column. PMID- 12568374 TI - Impact of normal-phase gradient elution in chiral chromatography: a novel, robust, efficient and rapid chiral screening procedure. AB - Novel normal-phase gradient systems have been employed for fast high-throughput chiral analyses of Discovery compounds in our research laboratories in Eli Lilly and Company. In this report, we describe an automated screening approach based on gradient elution, in order to achieve accurate enantiomeric excess determinations, and chiral separations when needed, in the shortest possible timeframe. Baseline resolution of enantiomers has been obtained for over 85% of the samples so tested. For the remaining cases, complete enantioseparation by isocratic optimisation is generally achieved in a single shot. This technique has been proven to be robust and is now standard operating procedure at our analytical research laboratories. PMID- 12568375 TI - Liquid chromatographic separation of phosphoramidate diastereomers on a polysaccharide-type chiral stationary phase. AB - To improve the therapeutic potential of anti-HIV nucleoside analogues (d4T, AZT, 3TC and ddl), the delivery of the corresponding monophosphate from neutral, membrane-permeable prodrugs has been realised by the synthesis of lipophilic phosphoramidate triester prodrugs, such as the simple phenyl-L-alaninephosphate derivatives. However, the present non-stereoselective synthesis results in a mixture of 1:1 diastereomers, which differ from the configuration of the phosphorus atom asymmetric center. Since each diastereomer may have different biological activity and pharmacokinetic profile, analytical methods have to be developed for their separation. This work aims at showing the ability of a polysaccharide-type chiral stationary phase to resolve such diastereomers in reversed-phase high-performance liquid chromatography. The influence of operating parameters has been studied to optimise the separation; a thermodynamic approach has also been investigated to gain an insight in the retention mechanism of the prodrugs. Preliminary validation study (linearity, accuracy, repeatability) has yielded good results; in addition, the feasibility of HPLC-electrospray-mass spectrometry (HPLC-ESI-MS) coupling has been demonstrated and it is expected that this will lead to lower detection limits. PMID- 12568376 TI - Nickel and copper complexes of a chelating methacrylate sorbent in the purification of chitinases and specific immunoglobulin G1 by immobilized metal ion affinity chromatography. AB - The isolation of the isoforms of endo- and exochitinases of Clostridium aminovalericum T1 and of the horseradish peroxidase (HRP)-specific immunoglobulin G1 from natural sources by immobilized metal ion affinity chromatography was studied. The effect of Cu2+ and Ni2+ complexes of iminodiacetic acid incorporated in porous glycidyl methacrylate-co-ethylene dimethacrylate and in agarose (Sepharose Fast Flow) beads on separation of the target polypeptides was analyzed. It was found that the Cu2+ complexes bound both the HRP-specific IgG1 and some isoforms of chitinases more strongly than the Ni2+ complexes. From the former complexes, both target polypeptides were eluted by a stepwise imidazole concentration gradient of 5-100 mM. The lower strength of Ni2+ complex binding with the HRP-specific IgG1 resulted in its easy elution with a pH gradient of 5.5 5 while some isoforms of chitinases required imidazole for their elution. The "fraction elution degree" of a target polypeptide (i.e., the ratio of its amounts in each eluate fraction and in the combined fractions) was used for the evaluation of the sorption selectivity and binding affinity of the separating components to the studied metal complexes. PMID- 12568377 TI - Protein partitioning in thermoseparating systems of a charged hydrophobically modified ethylene oxide polymer. AB - The phase behavior of a thermoseparating cationic hydrophobically modified ethylene oxide polymer (HM-EO) containing tertiary amines has been investigated at different pH, salt and sodium dodecyl sulfate (SDS) concentrations, in order to find a water/HM-EO two-phase system suitable for protein partitioning. The used polymer forms micellar aggregates that can be charged. By changing pH and SDS concentrations the netcharge of the SDS/HM-EO aggregate can be shifted from positive to negative. Bovine serum albumin (BSA) and lysozyme were partitioned in the thermoseparated two-phase systems of the cationic polymer at different pH, salt and SDS concentrations. The dominant attractive interactions between the polymer aggregates and the studied proteins were shown to be of electrostatic (Coulomb) nature rather than hydrophobic interaction. At low ionic strength the positively charged polymeric aggregates attracted negatively charged BSA and repelled positively charged lysozyme. Upon addition of SDS the negatively charged aggregates attracted lysozyme and repelled BSA. Thus, it was possible to direct proteins with different charges to the polymeric phase and redirect them to a polymer-depleted phase by changing the netcharge of the polymeric aggregates. The effect of different salts on the partitioning of BSA in a system of slightly positively charged HM-EO was studied. NaCl and KBr have a significant effect on driving the BSA to the polymer-depleted phase, whereas KF and K2SO4 have a smaller effect on the partitioning. The cloud point temperature of the charged polymer decreased upon addition of SDS near the isoelectric molar ratio of SDS to polymer and also upon salt addition. In the latter case the decrease was smaller than expected from model calculations based on Flory-Huggins theory, which were performed for a charged thermoseparating polymer at different charges and salt concentrations. PMID- 12568378 TI - Isocratic reversed-phase liquid chromatographic method for the simultaneous determination of (S)-methoprene, MGK264, piperonyl butoxide, sumithrin and permethrin in pesticide formulation. AB - An isocratic reversed-phase high-performance liquid chromatographic (RP-HPLC) method development for the simultaneous determination of five active ingredients (S)-methoprene, MGK264, PBO, sumithrin and permethrin in a new complex pesticide formulation is described. These active ingredients have similar polarities and contain isomers. The RP-HPLC method development approach began with the selection of a column based on the component structure information, bonded phase, and particle physical characteristics. Second, the mobile phase composition was changed to improve peak resolution and peak sensitivity, especially with analytes containing isomers. Choosing the match between the stationary phase and mobile phase composition, the developed RP-HPLC method not only can simplify the procedure appreciably but also significantly decrease total analysis time and increase peak height. The developed isocratic RP-HPLC method for the analysis of this new formulation was then validated for specificity, linearity, precision, and accuracy. The chromatographic peak identification was identified by LC-MS using the electrospray ionisation in the positive-ion mode. PMID- 12568379 TI - Pressurized liquid extraction of active ingredients (ginsenosides) from medicinal plants using non-ionic surfactant solutions. AB - The feasibility of employing aqueous non-ionic surfactant solutions as an alternative solvent system in pressurized liquid extraction (PLE) is demonstrated for the first time using the roots of American ginseng as model solid samples. When compared to the use of pure water or methanol, the presence of a common non ionic surfactant (Triton X-100) in water at a concentration above its critical micelle concentration was shown to enhance the amount of pharmacologically active ingredients (ginsenosides) extracted from ginseng roots. The advantages of using aqueous non-surfactant solutions were also demonstrated by comparing extraction performances between ultrasonic-assisted extraction and PLE methods. Furthermore, the combination of PLE and cloud point extraction was shown to be a new and effective approach for the rapid sample preconcentration of herbal materials prior to analysis by high-performance liquid chromatography. PMID- 12568380 TI - Retention behavior of metal particle dispersions in aqueous and nonaqueous carriers in thermal field-flow fractionation. AB - Until quite recently, theories on thermophoresis of particles predicted very low thermophoretic velocities of metal particles in liquids. This prediction was based on the understanding that the very high thermal conductivities of metals relative to most liquid media resulted in quite low temperature gradients across the metal particle thereby leading to low net force on the particle. In this paper, we report the retention behavior of submicrometer size metal particles of silver (Ag), gold (Au), palladium (Pd) and platinum (Pt) suspended in both aqueous and organic (specifically, acetonitrile and tetrahydrofuran) carrier liquids in thermal field-flow fractionation (ThFFF). The dependence of the metal particle retention on various factors such as particle composition, amount of added electrolyte, carrier liquid composition, field strength, channel thickness, and carrier flow-rate is evaluated and discussed. A comparison in particle retention behavior among equal-sized metal, latex and silica particles is also provided. PMID- 12568381 TI - Time-resolved determination of surface diffusion coefficients for physically adsorbed or chemisorbed species on heterogeneous surfaces, by inverse gas chromatography. AB - A new simple method is developed for measuring surface diffusion coefficients Ds of gases adsorbed on heterogeneous surfaces, using the reversed-flow version of inverse gas chromatography. The Ds values are found in a time-resolved way, together with the corresponding adsorption energy values, the local adsorbed concentrations, and the local adsorption isotherm values. A relative dynamic adsorption rate constant, an adsorption/desorption rate constant, and a surface reaction rate constant are also found in the same experiment, together with the total diffusion coefficient of the gas in the solid bed. The method has been applied for carbon monoxide, oxygen gas, and carbon dioxide as adsorbates on 75% Pt+25% Rh catalyst supported on SiO2, at 593.8 K. PMID- 12568382 TI - Comprehensive two-dimensional gas chromatography of volatile and semi-volatile components using a diaphragm valve-based instrument. AB - A high-temperature configuration for a diaphragm valve-based gas chromatography (GCXGC) instrument is demonstrated. GCxGC is a powerful instrumental tool often used to analyze complex mixtures. Previously, the temperature limitations of valve-based GCxGC instruments were set by the maximum operating temperature of the valve, typically 175 degrees C. Thus, valve-based GCxGC was constrained to the analysis of mainly volatile components; however, many complex mixtures contain semi-volatile components as well. A new configuration is described that extends the working temperature range of diaphragm valve-based GCxGC instruments to significantly higher temperatures, so both volatile and semi-volatile compounds can be readily separated. In the current investigation, separations at temperatures up to 250 degrees C are demonstrated. This new design features both chromatographic columns in the same oven with the valve interfacing the two columns mounted in the side of the oven wall so the valve is both partially inside as well as outside the oven. The diaphragm and the sample ports in the valve are located inside the oven while the temperature-restrictive portion of the valve (containing the O-rings) is outside the oven. Temperature measurements on the surface of the valve indicate that even after a sustained oven temperature of 240 degrees C, the portions of the valve directly involved with the sampling from the first column to the second column track the oven temperature to within 1.2% while the portions of the valve that are temperature-restrictive remain well below the maximum temperature of 175 degrees C. A 26-component mixture of alkanes, ketones, and alcohols whose boiling points range from 65 degrees C (n hexane) to 270 degrees C (n-pentadecane) is used to test the new design. Peak shapes along the first column axis suggest that sample condensation or carry-over in the valve is not a problem. Chemometric data analysis is performed to demonstrate that the resulting data have a bilinear structure. After over 6 months of use and temperature conditions up to 265 degrees C, no deterioration of the valve or its performance has been observed. PMID- 12568383 TI - Headspace microdrop analysis--an alternative test method for gasoline diluent and benzene, toluene, ethylbenzene and xylenes in used engine oils. AB - The primary standard test method used for the determination of gasoline diluent in used engine oils is method D 3525-93 of the American Society for Testing and Materials (ASTM), which involves direct injection of used oil onto a packed GC column and flame ionization detection. Recently, we have utilized a new headspace sampling method: headspace solvent microextraction (HSM), for GC and GC-MS analysis of gasoline diluent in used engine oils. High resolution capillary columns can be used without the necessity for the use of inlet cryogenic cooling or expensive sampling interfaces. This analytical method, which we generically refer to as headspace microdrop analysis yields results comparable to those obtained using the ASTM method, with the added benefit that it allows the quantification of individual volatile diluent components, including benzene, toluene, ethylbenzene and the xylenes. PMID- 12568384 TI - Phase-transfer catalytic determination of phenols as methylated derivatives by gas chromatography with flame ionization and mass-selective detection. AB - A convenient method for the GC determination of phenols as methylated derivatives is proposed, taking advantage of the beneficial features of phase-transfer catalysis (PTC). The optimal experimental conditions of pH, temperature, organic solvent, time of extraction-derivatization and amounts of the participating reactants and catalysts, were properly established. Several catalysts in soluble or polymer-bound form were tested. Most of them demonstrated appreciably high performance characteristics but the polymer-bound catalyst is most favourable due to its facile separation from the rest of the reaction system after the extraction-derivatization. Interferences with the extraction and derivatization yield were not noticed. The chromatographic separation of 11 methylated derivatives of phenols was complete within 23 min. The detection limits of the method, which range from 0.005 to 0.120 microg, are inadequate for drinking water analysis. However, the method was successfully applied to the analysis of fortified composite lake water samples using GC-flame ionization detection and GC MS in the single ion monitoring mode with the most abundant characteristic ions. Spiked recoveries of phenolics were in the range 94-102%, on the basis of distilled water calibration graph, signifying that PTC determination of phenols is not affected by the composition of such matrices. PMID- 12568385 TI - Methods for selective determination of persistent organochlorine pesticide residues in water and sediments by capillary gas chromatography and electron capture detection. AB - Different extraction methods were evaluated for the determination of fifteen organochlorine pesticides (OCPs) in water and sediments. Liquid-liquid extraction (LLE) was evaluated for the pesticides analyses in water while Soxhlet extraction (SE) and microwave assisted extraction (MAE) methods were compared in sediment. Of all the extracting solvents used, dichloromethane gave the best results. Percentage recoveries ranged from 71.03 +/- 8.15 (dieldrin) to 101.25 +/- 2.17% [a-benzenehexachloride (alpha-BHC)] in water with LLE. In sediments the percentage recoveries with Soxhlet extraction method varied between 88.22 +/- 7.85 (endrin) and 109.63 +/- 5.10% (beta-BHC) and ranged from 74.11 +/- 9.82 (2,4 DDT) to 97.50 +/- 4.56% (alpha-BHC) with MAE. The limits of detection for the OCPs ranged from 5.5 to 20.6 ng/l and between 0.6 and 2.1 ng/g. respectively. The LLE and the SE methods were applied to water and sediments samples, respectively, from marine and freshwater sources in the Eastern Cape Province of South Africa that receive runoffs from agricultural lands and effluents from industries. The levels of OCPs ranged from 5.5 (2,4-DDD) to 450 +/- 0.10 ng/l (beta-BHC) in water samples and from 0.6 (aldrin and 2,4-DDD) to 184 +/- 0.12 ng/g (beta-BHC) in sediments for triplicate analyses. Some endocrine disrupting OCPs such as DDT, DDE, heptachlor, endosulphan and the chlordanes were detected. PMID- 12568386 TI - Gas chromatography-mass spectrometry determination of the migration of phthalate plasticisers from polyvinyl chloride toys and childcare articles. AB - Two laboratory-based linear horizontal agitation methods for determining a range of phthalate esters from soft polyvinyl chloride (PVC) toys are presented in compliance with EU legislation. Both of these methods were validated through interlaboratory trials using a PVC reference disc and four soft PVC toy/childcare articles intended or likely to be mouthed. Two of these commercial samples contained diisononyl phthalate (DINP), one diisodecyl phthalate (DIDP) and one bis(2-ethylhexyl) phthalate (DEHP). Acceptable repeatability (r, within laboratory) and reproducibility (R, between-laboratory) data were demonstrated for both the analytical detection technique (GC-MS) (r = 9.8% and R = 8.1%) and agitation/extraction procedure (r=21.9% and R = 35.3% at 37 degrees C; r = 22.7% and R = 31.1% at 65 degrees C) for DINP. This was achieved through the participation of six laboratories. The remaining three phthalates from the EU Scientific Committee for Toxicity, Ecotoxicity and the Environment (CSTEE) list- dibutyl phthalate (DBP), benzyl butyl phthalate (BBP) and di-n-octyl phthalate (DnOP)--were not tested due to the unavailability of suitable materials. PMID- 12568387 TI - Role of buffer concentration and applied voltage in obtaining a good separation in planar electrochromatography. AB - Planar electrochromatography is performed by applying an electric field across a thin layer chromatography (TLC) plate. In addition to electroosmotic flow in the axial direction, there is also flow to the surface of the TLC layer, and this can substantially degrade the quality of separation. This effect is offset by Joule heating which causes evaporation of liquid from the layer surface, and which under some conditions causes degradation of separation quality by excessive drying of the layer. It is shown that pH, buffer concentration, and applied voltage control the balance between liquid being driven to the surface and liquid evaporating from the surface due to Joule heating. Conditions are discussed which result in good separation quality, or in separations degraded by either excessive wetting or drying of the layer. The above separations were performed at constant voltage. A chromatogram is presented that shows that a good separation is also obtained at constant power, i.e. under conditions where there is a constant amount of Joule heating. PMID- 12568388 TI - Electrochromatography with a 2.7 mm inner diameter monolithic column. AB - Monolithic columns of 2.7 mm I.D. have been prepared and used in electrochromatography (EC) separation. Although capillary electrochromatography (CEC) has higher separation efficiency, it displays some shortcomings, such as limited sample loadability and restricted concentration detectability etc. In this paper, we investigate the feasibility of EC separation with millimeter diameter monolithic columns. By using a designed preparation method of monolithic column packed with about 150 microm quartz sand, the effect of Joule heating can be reduced, and the processes of frit making and column packing can be avoided. The concentration detectability of the EC is improved comparing with that of CEC. Moreover, the separation efficiency of 52,000 plates/m was achieved with a 70 mm length and 2.7 mm I.D. monolithic column. PMID- 12568389 TI - Capillary zone electrophoresis of soil humic acid fractions obtained by coupling size-exclusion chromatography and polyacrylamide gel electrophoresis. AB - Capillary zone electrophoresis (CZE) was used for characterisation of soil humic acid (HA) fractions obtained by coupling size-exclusion chromatography with polyacrylamide gel electrophoresis, on the basis of their molecular size and electrophoretic mobility. CZE was conducted using several low alkaline buffers as background electrolyte (BGE): 50 mM carbonate, pH 9.0; 50 mM phosphate, pH 8.5; 50 mM borate, pH 8.3; 50 mM Tris-borate+1 mM EDTA+7 M urea+0.1% sodium dodecyl sulphate (SDS), pH 8.3. Independently of BGE conditions, the effective electrophoretic mobility of HA fractions were in good agreement with their molecular size. The better resolution of HA were obtained in Tris-borate-EDTA buffer with urea and SDS. This results indicated that CZE, mostly with BGE contained disaggregating agents, is useful for separating HAs in fractions with different molecular sizes. PMID- 12568390 TI - Matrix solid-phase dispersion for the liquid chromatographic determination of phenolic acids in Melissa officinalis. AB - Matrix solid-phase dispersion (MSPD) was used for sample preparation of plant material (Melissa officinalis, Lemon Balm) prior to liquid chromatography of rosmarinic, caffeic and protocatechuic acids, phenolic compounds present in this herb. Different MSPD sorbents and various elution agents were tested and the optimal extraction conditions determined with the aim to obtain extraction recoveries greater than 90% for all analytes. PMID- 12568391 TI - Column chromatographic separation of uranium(VI) and other elements using poly(dibenzo-18-crown-6) and ascorbic acid medium. AB - A selective and very effective separation method for uranium(VI) has been developed by using poly(dibenzo-18-crown-6) and column chromatography. The separations are carried out from ascorbic acid medium. The adsorption of uranium(VI) was quantitative from 0.00002 to 0.006 M ascorbic acid. The elution of uranium(VI) was quantitative with 2.0-8.0 M HCl and 2.0-5.0 M H2SO4. The capacity of poly(dibenzo-18-crown-6) for uranium(VI) was found to be 0.92 +/- 0.01 mmol g(-1) of crown polymer. Uranium(VI) was separated from a number of cations in binary as well as in multicomponent mixtures. The method was extended to the determination of uranium in geological samples. It is possible to separate and determine 5 ppm of uranium(VI) by this method. The method is very simple, rapid, selective and has good reproducibility (approximately +/- 2%). PMID- 12568392 TI - Simultaneous determination of long-chain aliphatic aldehydes and waxes in olive oils. AB - A procedure for the simultaneous determination of long-chain aliphatic aldehydes, and aliphatic and triterpenic waxes in virgin olive oils is described. A fraction containing these compounds was isolated from the oil using solid-phase extraction on silica-gel cartridges. The fraction was analyzed by capillary GC on 35% dimethyl-65%-diphenylpolysiloxane phase using on-column injection. In extra virgin olive oils, the long-chain aliphatic aldehydes with even carbon atom numbers from C22 to C30 were identified by comparison of retention times and mass spectra with those of synthesized standards. The concentration of total aldehydes ranged from 20.2 to 108.0 mg/kg-n-hexacosanal being the most abundant aldehyde. The determination of aliphatic waxes was achieved with similar or better precision than that of the EU official methods. PMID- 12568393 TI - Congenic strains of mice for verification and genetic decomposition of quantitative trait loci for femoral bone mineral density. AB - Peak femoral volumetric bone mineral density (femoral bone mineral density) in C57BL/6J (B6) 4-month-old female mice is 50% lower than in C3H/HeJ (C3H) and 34% lower than in CAST/EiJ (CAST) females. Genome-wide analyses of (B6 x C3H)F2 and (B6 x CAST)F2 4-month-old female progeny demonstrated that peak femoral bone mineral density is a complex quantitative trait associated with genetic loci (QTL) on numerous chromosomes (Chrs) and with trait heritabilities of 83% (C3H) and 57% (CAST). To test the effect of each QTL on femoral bone mineral density, two sets of loci (six each from C3H and CAST) were selected to make congenic strains by repeated backcrossing of donor mice carrying a given QTL-containing chromosomal region to recipient mice of the B6 progenitor strain. At the N6F1 generation, each B6.C3H and B6.CAST congenic strain (statistically 98% B6-like in genomic composition) was intercrossed to obtain N6F2 progeny for testing the effect of each QTL on femoral bone mineral density. In addition, the femoral bone mineral density QTL region on Chr 1 of C3H was selected for congenic subline development to facilitate fine mapping of this strong femoral bone mineral density locus. In 11 of 12 congenic strains, 6 B6.C3H and 5 B6.CAST, femoral bone mineral density in mice carrying c3h or cast alleles in the QTL regions was significantly different from that of littermates carrying b6 alleles. Differences also were observed in body weight, femoral length, and mid-diaphyseal periosteal circumference among these 11 congenic strains when compared with control littermates; however, these latter three phenotypes were not consistently correlated with femoral bone mineral density. Analyses of eight sublines derived from the B6.C3H-1T congenic region revealed two QTLs: one located between 36.9 and 49.7 centiMorgans (cM) and the other located between 73.2 and 100.0 cM distal to the centromere. In conclusion, these congenic strains provide proof of principle that many QTLs identified in the F2 analyses for femoral bone mineral density exert independent effects when transferred and expressed in a common genetic background. Furthermore, significant differences in femoral bone mineral density among the congenic strains were not consistently accompanied by changes in body weight, femur length, or periosteal circumference. Finally, decomposition of QTL regions by congenic sublines can reveal additional loci for phenotypes assigned to a QTL region and can markedly refine genomic locations of quantitative trait loci, providing the opportunity for candidate gene testing. PMID- 12568394 TI - From QTL mapping to genes: the long and winding road. PMID- 12568395 TI - Mechanisms of tumor metastasis to the bone: challenges and opportunities. AB - In human cancers, bone is a common site for metastasis. It is well known that metastasis is the cause of morbidity and mortality in patients with cancer. Both breast and prostate carcinomas have a propensity to metastasize to bone. In general, metastatic breast cancers result in osteolytic lesions. On the other hand, prostate cancer metastases are osteoblastic and result in osteosclerosis. Thus, bone formation and bone resorption are at the crux of the cancer metastasis problem. For example, in the prostate, there is a vicious cycle of metastasis to bone (Fig. 1). Metastases to bone causes excruciating bone pain, pathological fractures, and eventually death, and therefore is a serious challenge to both bone biologists and cancer cell biologists. The stromal-epithelial interactions in breast and prostate are critical in initiation of carcinogenesis and the progression of the metastatic cascade to bone (Fig. 2). Over a hundred years ago, Stephen Paget enunciated the seed and soil hypothesis in which seeds of metastatic cancer cells of breast preferentially settle in the soil of bone matrix. Thus, the prostate/breast cancer bone interface and continuum has continuously presented challenges and opportunities and were discussed at a recent workshop. PMID- 12568396 TI - A proteome study of secreted prostatic factors affecting osteoblastic activity: galectin-1 is involved in differentiation of human bone marrow stromal cells. AB - Prostate cancer cells metastasize to bone causing a predominantly osteosclerotic response. It has been shown that cells from the human prostate cancer cell line PC3 secrete factors that influence the behavior of osteoblast-like cells. Some of these factors with mitogenic activity have been found to be proteins with molecular weights between 20 and 30 kDa, but the identity of the osteoblastic mitogenic factor or factors produced by prostate cancer cells is still unknown. Therefore, the aim of this study was to characterize the protein profile of conditioned medium (CM) from PC3 cells in the molecular weight range from 5 to 30 kDa using proteome analysis. A protein profile of the CM from PC3 cells was performed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Thirty protein spots with molecular weights ranging from 5 to 30 kDa were analyzed by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). One of these spots was identified as galectin-1. We examined whether PC3 CM, recombinant galectin-1 alone, or combined with insulin-like growth factor-I (IGF-I) had any effects on the proliferation or differentiation of human bone marrow stromal (hBMS) cells. Furthermore, we tested whether adhesion of PC3 cells to plastic, laminin, fibronectin, and collagen type I was influenced by lactose, which inhibits galectin-1. Galectin-1 (1000 ng/ml) inhibited the proliferation of hBMS cells up to 70 +/- 12% (treated/control) of control in contrast to PC3 CM, which induced hBMS cell proliferation by 3-fold. This effect was abolished by IGF-I. PC3 CM and galectin-1 in concentrations of 10 and 1000 ng/ml increased the alkaline phosphatase (ALP) activity of hBMS cells up to 175 +/- 27%, 137 +/- 8%, and 131 +/- 11%, respectively, compared with ALP activity of untreated cells, and inhibited the secretion of osteocalcin (OC) up to 81 +/- 3%, 93 +/- 1%, and 58 +/- 2%, respectively, compared with OC secretion of untreated cells. These effects were affected by IGF-I. Lactose inhibited adhesion of PC3 cells to plastic, fibronectin, laminin, and collagen type I up to 58 +/- 4%, 30 +/- 12, 72 +/- 9%, and 86 +/- 4%. In conclusion, galectin-1 modulated osteoblastic proliferation and differentiation. These effects were affected by IGF-I. Thus, galectin-1 is likely be involved in the osteoblastic response, caused by prostate cancer cells metastasizing into bone, by affecting the matrix mineralization. PMID- 12568397 TI - Antagonistic effects of different classes of bisphosphonates in osteoclasts and macrophages in vitro. AB - Nitrogen-containing bisphosphonates, such as alendronate and ibandronate, inhibit bone resorption by preventing protein prenylation in osteoclasts, whereas non nitrogen-containing bisphosphonates, such as clodronate, are metabolized to nonhydrolyzable analogs of ATP, resulting in osteoclast apoptosis. Because these two classes of bisphosphonates have different molecular mechanisms of action, we examined in vitro whether combined treatment with clodronate and alendronate would alter antiresorptive effectiveness. Although, in cultures of rabbit osteoclasts, the antiresorptive effect of 10 microM alendronate was increased by the addition of clodronate, the effect of higher concentrations of alendronate was not altered by addition of clodronate. Furthermore, the inhibition of protein prenylation in osteoclasts caused by higher alendronate concentrations was partially prevented by cotreatment with clodronate. As in osteoclasts, the inhibition of protein prenylation in J774 cells caused by alendronate or ibandronate treatment was dose-dependently prevented by cotreatment with clodronate. Furthermore, alendronate-induced J774 apoptosis was significantly inhibited in the presence of clodronate. The presence of clodronate also decreased the short-term cellular uptake of [14C]ibandronate. These observations suggest that combined treatment with clodronate could enhance the antiresorptive effect of a low concentration of nitrogen-containing bisphosphonate, but clodronate can also antagonize some of the molecular actions and effects of higher concentrations of nitrogen-containing bisphosphonates. The exact molecular basis for the antagonistic effects between bisphosphonates remain to be determined, but could involve competition for cellular uptake by a membrane-bound transport protein. PMID- 12568398 TI - Changes in Runx2/Cbfa1 expression and activity during osteoblastic differentiation of human bone marrow stromal cells. AB - Runx2/Cbfa1 has been identified as a "master gene" controlling osteoblast differentiation. However, its role in inducing the osteoblast phenotype has been characterized primarily in rodent systems. Thus, we examined Runx2/Cbfa1 messenger RNA, protein, and activity levels during osteoblastic differentiation of human bone marrow stromal (BMSC) cells. Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) analysis demonstrated that the expression of alkaline phosphatase and osteocalcin mRNAs increased in a time-dependent manner with the development of the osteoblast phenotype by these cells (hMS2-15). Type II Runx2/Cbfa1 messenger RNA was found to be constitutively expressed in hMS2-15 cells and not altered during differentiation; there was no detectable expression of the type I Runx2/Cbfa1 transcript. Interestingly, despite the absence of any change in Runx2/Cbfa1 messenger RNA levels during osteoblastic differentiation of these cells, the activity of Runx2/Cbfa1, as assessed by binding to the osteoblast-specific cis-acting element 2 (OSE2), increased markedly at all time points examined, with the highest activity level seen at day 7. Similar results were observed in primary cultures of less differentiated human marrow-derived mesenchymal stem cells. Immunoprecipitation and Western blot analysis revealed that whereas there was no increase in Runx2/Cbfa1 protein levels with differentiation in hMS2-15 cells, there was an increase in Runx2/Cbfa1 phosphorylation. Thus, in contrast to rodent systems where osteoblast differentiation is associated with increased synthesis of Runx2/Cbfa1, we find that in human BMSC, osteoblastic differentiation is associated primarily with increases in Runx2/Cbfa1 activity, without a change in messenger RNA or protein levels. Our findings also show that the increase in Runx2/Cbfa1 activity occurs through a posttranslational mechanism involving phosphorylation of key residues. PMID- 12568399 TI - Cathepsin K mRNA and protein expression in prostate cancer progression. AB - Prostate cancer (CaP) is the most commonly diagnosed malignancy in men and is often associated with bone metastases, which cause much of the morbidity associated with CaP. Lesions associated with CaP generally exhibit increased bone formation and resorption. Increased bone resorption may release factors from the extracellular matrix that contribute to tumor growth. Cathepsin K (cat K) is a cysteine protease that exhibits strong degradative activity against the extracellular matrix and is involved in osteoclast-mediated bone destruction. In this study, we analyzed the expression of cat K in CaP cell lines and patient samples. Cat K message was detected in CaP cell lines by reverse transcription polymerase chain reaction (RT-PCR) and in primary CaP and metastases by in situ hybridization. Immunohistochemistry revealed variable expression of cat K in primary CaP samples, as well as nonosseous metastases, whereas expression in bone metastases was significantly higher than in primary CaP, and normal prostate tissues were negative. Cat K protein was detected in CaP cell lines by Western blotting after immunoprecipitation. Cat K enzymatic activity was also detected in CaP cell lines by a fluorogenic assay and by an assay for degradation of collagen type I. Increased levels of NTx, a marker of bone matrix degradation mediated primarily by cat K, were also detected in sera of patients with CaP bone metastases. We hypothesize that CaP-expressed cat K may contribute to the invasive potential of CaP, while increased expression in bone metastases is consistent with a role in matrix degradation. PMID- 12568400 TI - Evaluation of bone mineral density and fat-lean distribution in patients with multiple myeloma in sustained remission. AB - To study the usefulness of bone mineral density (BMD) in the follow-up of myeloma (MM) patients, BMD was evaluated in 44 MM patients in sustained remission for at least 2 years (35.4 +/- 10.5 months) after high-dose or conventional chemotherapy in a retrospective study. Patients never received bisphosphonates before or during the follow-up. Patients underwent lumbar spine (LS) BMD and a whole body (WB) BMD testing before therapy and at least once in the remission period. At baseline, mean LS BMD was 0.863 +/- 0.026 g/cm2, mean lumbar Z-score was -1.45 SD. LS BMD significantly increased from baseline by 5 +/- 1.8%, 9.3 +/- 1.7%, and 14 +/- 1.9% at 1, 2, and 3 years, respectively. The percentage of patients with a T-score below 2.5 SD decreased from 39% at baseline to 18.5% at 3 years. Compared with baseline, WB BMD decreased by -2.8 +/- 0.5%, -2.6 +/- 0.7%, and -1.7 +/- 0.6% at 1, 2, and 3 years, respectively. Mean percentage change of the fat compartment increased from baseline by +28.4 +/- 7.1% at the trunk, and +17.1 +/- 5% in peripheral areas at 3 years. In conclusion, in MM patients in remission after chemotherapy, LS BMD progressively increased after a mean follow-up of 3 years. These patients never received bisphosphonates, so this increase was related to the anti-myeloma treatment. The major effect on BMD was observed at the LS, which is primarily composed of trabecular bone containing the bone marrow. Interestingly, a drastic increase of the fat content was also observed. These results underlined that BMD and fat-lean evaluation could be of interest in the follow-up of MM patients. PMID- 12568401 TI - Positive linear growth and bone responses to growth hormone treatment in children with types III and IV osteogenesis imperfecta: high predictive value of the carboxyterminal propeptide of type I procollagen. AB - Extreme short stature is a cardinal feature of severe osteogenesis imperfecta (OI), types III and IV. We conducted a treatment trial of growth hormone in children with OI and followed linear growth velocity, bone metabolism markers, histomorphometrics, and vertebral bone density. Twenty-six children with types III and IV OI, ages 4.5-12 years, were treated with recombinant growth hormone (rGH), 0.1-0.2 IU/kg per day for 6 days/week, for at least 1 year. Length, insulin-like growth factor (IGF-I), insulin-like growth factor binding protein (IGFBP-3), bone metabolic markers, and vertebral bone density by DXA were evaluated at 6-month intervals. An iliac crest biopsy was obtained at baseline and 12 months. Approximately one-half of the treated OI children sustained a 50% or more increase in linear growth over their baseline growth rate. Most responders (10 of 14) had moderate type IV OI. All participants had positive IGF I, IGFBP-3, osteocalcin, and bone-specific alkaline phosphatase responses. Only the linear growth responders had a significant increase in vertebral DXA z-score and a significant decrease in long bone fractures. After 1 year of treatment, responders' iliac crest biopsy showed significant increases in cancellous bone volume, trabecular number, and bone formation rate. Responders were distinguished from nonresponders by higher baseline carboxyterminal propeptide (PICP) values (p < 0.05), suggesting they have an intrinsically higher capacity for collagen production. The results show that growth hormone can cause a sustained increase in the linear growth rate of children with OI, despite the abnormal collagen in their bone matrix. In the first year of treatment, growth responders achieve increased bone formation rate and density, and decreased fracture rates. The baseline plasma concentration of PICP was an excellent predictor of positive response. PMID- 12568402 TI - Purification of matrix Gla protein from a marine teleost fish, Argyrosomus regius: calcified cartilage and not bone as the primary site of MGP accumulation in fish. AB - Matrix Gla protein (MGP) belongs to the family of vitamin K-dependent, Gla containing proteins, and in mammals, birds, and Xenopus, its mRNA was previously detected in extracts of bone, cartilage, and soft tissues (mainly heart and kidney), whereas the protein was found to accumulate mainly in bone. However, at that time, it was not evaluated if this accumulation originated from protein synthesized in cartilage or in bone cells because both coexist in skeletal structures of higher vertebrates and Xenopus. Later reports showed that MGP also accumulated in costal calcified cartilage as well as at sites of heart valves and arterial calcification. Interestingly, MGP was also found to accumulate in vertebra of shark, a cartilaginous fish. However, to date, no information is available on sites of MGP expression or accumulation in teleost fishes, the ancestors of terrestrial vertebrates, who have in their skeleton mineralized structures with both bone and calcified cartilage. To analyze MGP structure and function in bony fish, MGP was acid-extracted from the mineralized matrix of either bone tissue (vertebra) or calcified cartilage (branchial arches) from the bony fish, Argyrosomus regius, separated from the mineral phase by dialysis, and purified by Sephacryl S-100 chromatography. No MGP was recovered from bone tissue, whereas a protein peak corresponding to the MGP position in this type of gel filtration was obtained from an extract of branchial arches, rich in calcified cartilage. MGP was identified by N-terminal amino acid sequence analysis, and the resulting protein sequence was used to design specific oligonucleotides suitable to amplify the corresponding DNA by a mixture of reverse transcription-polymerase chain reaction (RT-PCR) and 5'rapid amplification of cDNA (RACE)-PCR. In parallel, ArBGP (bone Gla protein, osteocalcin) was also identified in the same fish, and its complementary DNA cloned by an identical procedure. Tissue distribution/accumulation was analyzed by Northern blot, in situ hybridization, and immunohistochemistry. In mineralized tissues, the MGP gene was predominantly expressed in cartilage from branchial arches, with no expression detected in the different types of bone analyzed, whereas BGP mRNA was located in bone tissue as expected. Accordingly, the MGP protein was found to accumulate, by immunohistochemical analysis, mainly in the extracellular matrix of calcified cartilage. In soft tissues, MGP mRNA was mainly expressed in heart but in situ hybridization, indicated that cells expressing the MGP gene were located in the bulbus arteriosus and aortic wall, rich in smooth muscle and endothelial cells, whereas no expression was detected in the striated muscle myocardial fibers of the ventricle. These results show that in marine teleost fish, as in mammals, the MGP gene is expressed in cartilage, heart, and kidney tissues, but in contrast with results obtained in Xenopus and higher vertebrates, the protein does not accumulate in vertebra of non-osteocytic teleost fish, but only in calcified cartilage. In addition, our results also indicate that the presence of MGP mRNA in heart tissue is due, at least in fish, to the expression of the MGP gene in only two specific cell types, smooth muscle and endothelial cells, whereas no expression was found in the striated muscle fibers of the ventricle. In light of these results and recent information on expression of MGP gene in these same cell types in mammalian aorta, it is likely that the levels of MGP mRNA previously detected in Xenopus, birds, and mammalian heart tissue may be restricted to regions rich in smooth muscle and endothelial cells. Our results also emphasize the need to re-evaluate which cell types are involved in MGP gene expression in other soft tissues and bring further evidence that fish are a valuable model system to study MGP gene expression and regulation. PMID- 12568403 TI - Expression of either NF-kappaB p50 or p52 in osteoclast precursors is required for IL-1-induced bone resorption. AB - Interleukin (IL)-1 is implicated in postmenopausal- and inflammation-mediated bone loss. Its expression is regulated by NF-kappaB and vice versa. To examine the role of NF-kappaB p50 and p52 (they are required for osteoclast formation during embryonic development) in IL-1-induced resorption, we used various NF kappaB knockout (KO) mice, including p50-/- and p52-/- single KO, p50-/- and p52+/- (3/4KO), and p50-/- and p52-/- double KO (dKO) mice. IL-1 increased blood calcium and bone resorption in wild-type (wt), p50, and p52 single KO mice, but not in 3/4KO or dKO mice. Osteoclast formation was impaired in bone marrow cultures from 3/4KO compared with single KO and wt mice treated with IL-1. IL-1 receptor expression was similar in colony forming unit-granulocyte macrophage (CFU-GM) colony cells from wt and dKO mice. However, IL-1 promoted CFU-GM colony formation and survival as well as the formation, activity, and survival of osteoclasts generated from these colonies from wt mouse splenocytes, but not from dKO splenocytes. No difference in expression of the osteoclast regulatory cytokines, RANKL, and OPG, was observed in osteoblasts from wt and dKO mice. Thus, expression of either NF-kappaB p50 or p52 is required in osteoclasts and their precursors, rather than osteoblasts, for IL-1-mediated bone resorption. PMID- 12568404 TI - Expression of RANKL and OPG correlates with age-related bone loss in male C57BL/6 mice. AB - Osteoblasts regulate the recruitment and activity of osteoclasts through expression of RANKL and osteoprotegerin (OPG). To determine whether expression of RANKL and OPG change with age and how these changes relate to the bone loss of aging, we measured bone mass and cancellous volume, and expression of RANKL, OPG, alkaline phosphatase (AP), osteocalcin (OC), and alpha I collagen (COLL) in whole bone and osteoblast-like cells in culture using 6-week- (young), 6-month- (adult), and 24-month-old (old) mice. Cancellous volume decreased by 20% from young to adult and by 52% from adult to old. RANKL mRNA levels in whole bone were 2.1-fold and 4.4-fold higher in adult and old mice, respectively, compared with young mice, whereas OPG mRNA levels decreased with age slightly. RANKL expression was negatively (r = -0.99) and OPG was positively (r = 0.92) correlated with cancellous bone volume. Expression of RANKL was higher and OPG lower in cells from older animals early in culture (day 7). With cell maturation, RANKL mRNA levels in cells from young and adult mice increased, whereas levels in cells from old animals decreased. By 21 and 28 days of culture, no differences were found in RANKL mRNA in osteoblast-like cells among different age groups. We conclude that expression of RANKL and OPG change with age in whole bone and in cultured osteoblast-like cells. These changes favor increased osteoclast over osteoblast activity, and may explain, in part, the imbalance in bone formation and resorption associated with aging. PMID- 12568405 TI - Overexpression of RANKL implicates IFN-beta-mediated elimination of B-cell precursors in the osteopetrotic bone of microphthalmic mice. AB - The microphthalmic (mi) mouse possesses a dominant negative mutation in the microphthalmia-associated transcript factor (MITF) transcription factor. These animals are characterized by reduced numbers of peripheral mast and natural killer (NK) cells, are osteopetrotic because of osteoclast reduction and malfunction, lack functional melanocytes, and are deficient for maturing B-cells within the bone marrow. Granulocyte precursor cells, however, are functionally maintained within the mi bone marrow. A central question has been whether the B cell deficiency of the mi mouse marrow is caused by the absence of an MITF controlled gene product or because of the compromised, osteopetrotic environment. In this report, we examined mi marrow by performing transcriptional mapping analyses of candidate genes whose products are instrumental for functional osteoclast and B-cell development. Surprisingly, the expression of a subset of such genes including RANKL, stromal-derived factor (SDF-1), B-cell lymphotactin chemokine (BLC), and RANK was dramatically enhanced in the mi marrow. Normal and mutant marrow were also analyzed by subtractive transcript cloning, which identified a number of known and unknown genes with altered transcriptional activity. One such unknown mouse gene possesses a human counterpart that is interferon-beta (IFN-beta) inducible, suggesting the osteopetrotic marrow is enriched for IFN-beta, a cytokine that is known to eliminate B-cell precursors. A model is proposed suggesting excess RANKL sets off a cascade of cytokine production including IFN-beta that leads to the preferential elimination of B cell precursors in the marrow of osteopetrotic marrow. PMID- 12568406 TI - Endoglin is expressed on human chondrocytes and forms a heteromeric complex with betaglycan in a ligand and type II TGFbeta receptor independent manner. AB - Previous work has implicated transforming growth factor beta (TGFbeta) as an essential mediator of cartilage repair and TGFbeta signaling as a requirement for the maintenance of articular cartilage in vivo. However, the mechanisms regulating TGFbeta action in chondrocytes are poorly understood. Endoglin, an accessory receptor of the TGFbeta receptor superfamily, is highly expressed on endothelial cells and has been shown to potently modulate TGFbeta responses. It is not known whether chondrocytes express endoglin or whether it modulates TGFbeta signaling in these cells. In this study, we show that endoglin is expressed on human chondrocytes at levels comparable with endothelial cells and that it forms higher order complexes with the types I and II TGFbeta receptors. More importantly, we show that endoglin forms a heteromeric complex with betaglycan on these cells at endogenous receptor concentrations and ratios. Endoglin complexes with betaglycan in a ligand-independent and -dependent manner as indicated by co-immunoprecipitation in the absence of TGFbeta and after affinity labeling with radiolabeled TGFbeta, respectively. Also, the endoglin betaglycan association can occur independently of the type II TGFbeta receptor. These findings, taken together with the available evidence that endoglin and betaglycan are potent modulators of TGFbeta signal transduction, imply that the complex formation between endoglin and betaglycan may be of critical significance in the regulation of TGFbeta signaling in chondrocytes. PMID- 12568407 TI - Deficient acquisition of bone during maturation underlies idiopathic osteoporosis in men: evidence from a three-generation family study. AB - To address the issue whether deficient acquisition of bone during maturation or adult-onset bone loss is primarily to blame for idiopathic osteoporosis in men, we assessed indices of bone mineral density and size, as well as biochemical markers of bone turnover in 61 probands (ages 20-65 years) with idiopathic osteoporosis (z-score < or = -2.0 at the spine or hip), their first-degree relatives (n = 130), and age-matched controls. There was no indication of accelerated bone loss. Indeed, in probands, the observed bone deficit versus controls was unrelated to the age of probands, and indices of bone turnover were not significantly different from controls. On the other hand, a specific deficit in bone acquisition was suggested by findings of lower bone mineral density values in three generations of male and female relatives of the probands, including their offspring; bone turnover in relatives was not different from controls. The bone mineral density deficit was more pronounced in male compared with female relatives; approximately 60% of the sons had a spinal bone mineral density z-score of less than -2.0. There also was a skeletal site-specificity in probands and their male relatives with a larger areal bone mineral density deficit at the spine compared with the hip and the forearm. The deficit at the spine corresponded to a reduction of both volumetric bone mineral density and bone size; a similar less pronounced deficit in volumetric bone mineral density, but not in bone size, was observed at the femoral neck. These findings in probands and their first-degree relatives point toward a major contributory role of a genetically determined maturational defect in bone acquisition in the pathogenesis of idiopathic osteoporosis in men. PMID- 12568408 TI - Relative contributions of bone density, bone turnover, and clinical risk factors to long-term fracture prediction. AB - Long-term fracture prediction using bone mineral density remains controversial, as does the additional contribution from assessing bone turnover or clinical risk factors. We measured bone mineral density at various sites, along with biochemical markers of bone turnover, sex steroid levels, and over 100 clinical variables, at baseline on an age-stratified sample of 304 Rochester, MN women in 1980. The 225 postmenopausal women were subsequently followed for 3146 person years (median, 16.2 years per subject), wherein they experienced 302 new fractures: 81% resulted from minimal or moderate trauma and 60% of these involved the proximal femur, thoracic or lumbar vertebrae, or distal forearm. Accounting for multiple fractures per subject, these osteoporotic fractures together were best predicted by baseline femoral neck bone mineral density (age-adjusted hazard ratio [HR] per SD decrease, 1.37; 95% CI, 1.10-1.70); 19 moderate trauma forearm fractures were best predicted by distal radius bone mineral content, whereas 28 hip fractures and 100 vertebral fractures were best predicted by femoral neck bone mineral density. Femoral neck bone mineral density performed comparably in predicting osteoporotic fracture risk within the first decade of follow-up (HR, 1.38; 95% CI, 1.10-1.74) as well as more than 10 years after baseline (HR, 1.39; 95% CI, 1.05-1.84). The older biochemical markers were not associated with fractures, but serum "free" estradiol index was independently predictive of short and long-term fracture risk. Consistent clinical risk factors were not identified, but statistical power was limited. Identifying patients at increased long-term risk of fracture is challenging, but it is reassuring that femoral neck bone mineral density can predict osteoporotic fractures up to 20 years later. PMID- 12568409 TI - Relation of androgen receptor gene polymorphism to bone mineral density and fracture risk in early postmenopausal women during a 5-year randomized hormone replacement therapy trial. AB - In women, the influence of androgens on bone health is not clear. It has been suggested that the androgen receptor (AR) genotype is associated with bone mineral density and serum androgen levels in pre- and perimenopausal women, but the association between AR genotype, bone mineral density, and fracture risk has not been studied in postmenopausal women. Therefore, we studied whether AR polymorphism affects bone mineral density, bone mineral density change, or fracture risk in a 5-year randomized hormone replacement therapy (HRT) trial on 331 early postmenopausal women (mean baseline age, 52.7 +/- 2.3 years). The participants consisted of two treatment groups: the HRT group (n = 151) received a sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate with or without vitamin D3, 100-300 IU + 93 mg calcium as lactate/day, and the non-HRT group (n = 180) received 93 mg calcium alone or in combination with vitamin D3, 100-300 IU/day for 5 years. Bone mineral density was measured from lumbar spine and proximal femur (DXA) before and after the 5-year trial. All new symptomatic, radiographically defined fractures were recorded during the follow up. The length of CAG repeat in exon 1 of AR gene was evaluated after polymerase chain reaction (PCR) amplification. The subjects were divided into three repeat groups according to AR alleles. None of the baseline characteristics were associated with AR gene polymorphism and HRT treatment. The polymorphism did not influence the calculated annual changes of lumbar or femoral neck bone mineral density during the 5-year follow-up in the HRT (p = 0.926 and 0.146, respectively) or non-HRT (p = 0.818 and 0.917, respectively) groups. In all, 28 women sustained 33 fractures during the follow-up. Thus, the numbers of fractures were limited. The AR repeat length variation was not significantly associated with fracture risk in the HRT or non-HRT groups (p = 0.632 and 0.459, respectively; Cox proportional hazards model). In conclusion, AR gene polymorphism was not associated with baseline bone mineral density, 5-year bone mineral density change, or fracture risk in early postmenopausal Finnish women. PMID- 12568410 TI - Ten-year longitudinal relationship between physical activity and lumbar bone mass in (young) adults. AB - Little is known about the influence of long-term daily physical activity (PA) on lumbar bone mass after peak bone mass has been reached, that is, during [young] adulthood. The purpose of this study was to investigate the longitudinal relationship between PA and lumbar bone mineral density (LBMD) in healthy subjects over a 10-year period. The data reported here relate to 225 male and 241 female participants in the Amsterdam Growth and Health Longitudinal Study, who were measured at the mean ages of 27, 32, and/or 36. LBMD, habitual daily PA, total body weight, and calcium intake were assessed at each measurement point. The effects of two aspects of PA were analyzed: the mechanical (MECHPA; sum of all ground reaction forces) and metabolic (METPA; weighted metabolic score of intensity, frequency, and duration) components, each within a separate model. Multilevel analysis was used to investigate the relationship between PA and LBMD over the 10-year period. Gender, total body weight, and calcium intake were included in the analysis as covariates. The results indicated that MECHPA was a significant positive linear predictor of LBMD for males (r = 0.09; p < 0.001) but not for females. For the METPA, no linear longitudinal relationship with LBMD was found. The results suggest that there is a metabolic threshold at which extra PA becomes "deleterious" and METPA in its totality becomes ineffective for LBMD. It is concluded that during the (young) adult period, between 27 and 36 years of age, PA causing mechanical loading on the skeleton has a small positive influence on LBMD in males. PMID- 12568411 TI - Hormone replacement therapy dissociates fat mass and bone mass, and tends to reduce weight gain in early postmenopausal women: a randomized controlled 5-year clinical trial of the Danish Osteoporosis Prevention Study. AB - The aim of this study was to study the influence of hormone replacement therapy (HRT) on weight changes, body composition, and bone mass in early postmenopausal women in a partly randomized comprehensive cohort study design. A total of 2016 women ages 45-58 years from 3 months to 2 years past last menstrual bleeding were included. One thousand were randomly assigned to HRT or no HRT in an open trial, whereas the others were allocated according to their preferences. All were followed for 5 years for body weight, bone mass, and body composition measurements. Body weight increased less over the 5 years in women randomized to HRT (1.94 +/- 4.86 kg) than in women randomized to no HRT (2.57 +/- 4.63, p = 0.046). A similar pattern was seen in the group receiving HRT or not by their own choice. The smaller weight gain in women on HRT was almost entirely caused by a lesser gain in fat. The main determinant of the weight gain was a decline in physical fitness. Women opting for HRT had a significantly lower body weight at inclusion than the other participants, but the results in the self-selected part of the study followed the pattern found in the randomized part. The change in fat mass was the strongest predictor of bone changes in untreated women, whereas the change in lean body mass was the strongest predictor when HRT was given. Body weight increases after the menopause. The gain in weight is related to a decrease in working capacity. HRT is associated with a smaller increase in fat mass after menopause. Fat gain protects against bone loss in untreated women but not in HRT treated women. The data suggest that women's attitudes to HRT are more positive if they have low body weight, but there is no evidence that the conclusions in this study are skewed by selection bias. PMID- 12568412 TI - Effects of vitamin D and calcium supplementation on falls: a randomized controlled trial. AB - Specific receptors for vitamin D have been identified in human muscle tissue. Cross-sectional studies show that elderly persons with higher vitamin D serum levels have increased muscle strength and a lower number of falls. We hypothesized that vitamin D and calcium supplementation would improve musculoskeletal function and decrease falls. In a double-blind randomized controlled trial, we studied 122 elderly women (mean age, 85.3 years; range, 63 99 years) in long-stay geriatric care. Participants received 1200 mg calcium plus 800 IU cholecalciferol (Cal+D-group; n = 62) or 1200 mg calcium (Cal-group; n = 60) per day over a 12-week treatment period. The number of falls per person (0, 1, 2-5, 6-7, >7 falls) was compared between the treatment groups. In an intention to treat analysis, a Poisson regression model was used to compare falls after controlling for age, number of falls in a 6-week pretreatment period, and baseline 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D serum concentrations. Among fallers in the treatment period, crude excessive fall rate (treatment - pretreatment falls) was compared between treatment groups. Change in musculoskeletal function (summed score of knee flexor and extensor strength, grip strength, and the timed up&go test) was measured as a secondary outcome. Among subjects in the Cal+D-group, there were significant increases in median serum 25 hydroxyvitamin D (+71%) and 1,25-dihydroxyvitamin D (+8%). Before treatment, mean observed number of falls per person per week was 0.059 in the Cal+D-group and 0.056 in the Cal-group. In the 12-week treatment period, mean number of falls per person per week was 0.034 in the Cal+D-group and 0.076 in the Cal-group. After adjustment, Cal+D-treatment accounted for a 49% reduction of falls (95% CI, 14 71%; p < 0.01) based on the fall categories stated above. Among fallers of the treatment period, the crude average number of excessive falls was significantly higher in the Cal-group (p = 0.045). Musculoskeletal function improved significantly in the Cal+D-group (p = 0.0094). A single intervention with vitamin D plus calcium over a 3-month period reduced the risk of falling by 49% compared with calcium alone. Over this short-term intervention, recurrent fallers seem to benefit most by the treatment. The impact of vitamin D on falls might be explained by the observed improvement in musculoskeletal function. PMID- 12568413 TI - Effect of long-term impact-loading on mass, size, and estimated strength of humerus and radius of female racquet-sports players: a peripheral quantitative computed tomography study between young and old starters and controls. AB - Bone characteristics of the humeral shaft and distal radius were measured from 64 female tennis and squash players and their 27 age-, height-, and weight-matched controls with peripheral quantitative tomography (pQCT) and dual energy X-ray absorptiometry (DXA). The players were divided into two groups according to the starting age of their tennis or squash training (either before or after menarche) to examine the possible differences in the loading-induced changes in bone structure and volumetric density. The following pQCT variables were used: bone mineral content, total cross-sectional area of bone (TotA), cross-sectional area of the marrow cavity (CavA) and that of the cortical bone (CoA), cortical wall thickness (CWT), volumetric density of the cortical bone (CoD) and trabecular bone (TrD), and torsional bone strength index for the shaft (BSIt) and compressional bone strength index for the bone end (BSIc). These bone strength indices were compared with the DXA-derived areal bone mineral density (aBMD) to assess how well the latter represents the effect of mechanical loading on apparent bone strength. At the humeral shaft, the loaded arm's greater bone mineral content (an average 19% side-to-side difference in young starters and 9% in old starters), was caused by an enlarged cortex (CoA; side-to-side differences 20% and 9%, respectively). The loaded humerus seemed to have grown periosteally (the CavA did not differ between the sites), leading to 26% and 11% side-to-side BSIt differences in the young and old starters, respectively. CoD was equal between the arms (-1% difference in both player groups). The side-to-side differences in the young starters' bone mineral content, CoA, TotA, CWT, and BSIt were 8-22% higher than those of the controls and 8-14% higher than those of the old starters. Old starters' bone mineral content, CoA, and BSIt side-to-side differences were 6-7% greater than those in the controls. The DXA-derived side-to side aBMD difference was 7% greater in young starters compared with that of the old starters and 14% compared with that in controls, whereas the difference between old starters and controls was 6%, in favor of the former. All these between-group differences were statistically significant. At the distal radius, the player groups differed significantly from controls in the side-to-side bone mineral content, TrD, and aBMD differences only: the young starters' bone mineral content difference was 9% greater, TrD and aBMD differences were 5% greater than those in the controls, and the old starters' TrD and aBMD differences were both 7% greater than those in the controls. In summary, in both of the female player groups, the structural adaptation of the humeral shaft to long-term loading seemed to be achieved through periosteal enlargement of the bone cortex, although this adaptation was clearly better in the young starters. Exercise-induced cortical enlargement was not so clear at the distal radius (a trabecular bone site), and the study suggested that at long bone ends, the trabecular density could be a modifiable factor to built a stronger bone structure. Conventional DXA based aBMD measurement detected the intergroup differences in the exercise induced bone gains, although, because it measured two dimensions of bone only, it seemed to underestimate the effect of exercise on the apparent bone strength, especially if the playing had been started during the growing years. PMID- 12568414 TI - Distinct anabolic response of osteoblast to low-intensity pulsed ultrasound. AB - Low-intensity pulsed ultrasound, a form of mechanical energy transmitted as high frequency acoustical pressure waves, provides noninvasive therapeutic treatment for accelerating fracture repair and distraction osteogenesis. Relatively young osteoblasts respond to ultrasound by transiently upregulating message levels of immediate-early genes as well as that of osteocalcin and insulin-like growth factor I (IGF-I). Osteocytes derived from newborn rat tibia and calvaria responded to a lesser extent only in c-fos and cyclooxygenase-2 (COX-2) messages. Compared with the stretched osteocytes, which use stretch-activated and parathyroid hormone (PTH)-potentiated Ca2+ influx as an entry route to the protein kinase A (PKA) signal transduction pathways, there was no evidence of Ca2+ internalization by any of the cells tested on exposure to the ultrasound. On the other hand, inhibitors of p38 mitogen-activated protein kinase (MAPK) and upstream phosphoinositide 3-kinase (PI3K) blocked COX-2 and osteocalcin upregulation by the ultrasound-exposed ST2, murine bone marrow-derived cells. This is distinct from the aforementioned osteocytic response to low-frequency stretching and implies the involvement of integrins. Our findings suggested that accelerated fracture repair and distraction osteogenesis by the low-intensity pulsed ultrasound depend, at least in part, on the stimulation of osteoblastic cells at relatively early stages of osteogenic lineage. Bone is under control of multiple regulatory mechanisms so that diverse physical forces can be reflected to the microenvironment of each cell, in turn, to the entire bone. PMID- 12568415 TI - Bone loss detection in rats using a mouse densitometer. AB - Estrogen-depletion bone-loss studies often use ovariectomized (ovx) rats and measure bone mineral density in vivo or ex vivo using DXA. Recently, a portable densitometer (PIXImus) was developed for mouse research; however, its use in rats is unclear. This study compared the ability of PIXImus and a standard densitometer (DPXL) to detect ovx-induced bone loss in rats both in vivo and ex vivo. Additionally, instrument accuracy was assessed by comparing measured bone mass with ash weight. Finally, the use of two distal femur regions of interest (ROI) to detect ovx-induced bone loss was evaluated. Twenty-three 6-month-old nulliparous female Sprague-Dawley rats were randomly assigned to sham or ovx groups. Distal femur bone mineral density was assessed at baseline and at 1 and 2 months postoperatively, using a PIXImus and DPXL densitometer. At 3 months postoperatively, all animals were killed, and ex vivo femur scans obtained. Distal femur bone loss was demonstrable by 1 month post-ovx using either densitometer. With the PIXImus, a 4-mm ROI demonstrated greater bone loss (p < 0.05) than an 8-mm ROI. Using the 4-mm ROI, similar amounts of bone loss were detected by the PIXImus and DPXL: 22.2% and 22.4%, respectively, at 2 months post ovx. Total femur bone mineral content was overestimated by the PIXImus but highly correlated with the DPXL measurement (r = 0.988) and ash weight (r = 0.998). Given its comparability to standard DXA plus its rapid scan speed and portability, the PIXImus is useful in evaluating ovx-induced osteopenia in rats. PMID- 12568416 TI - Identification of a novel tandem duplication in exon 1 of the TNFRSF11A gene in two unrelated patients with familial expansile osteolysis. AB - Familial expansile osteolysis (FEO) is a rare autosomal dominant disorder characterized by striking focal expansile osteolytic bone lesions and generalized osteopenia, often accompanied by characteristic early hearing loss and dental disease. The TNFRSF11A gene encodes the receptor activator of nuclear factor kappaB (RANK), which has been demonstrated to be essential in bone remodeling and osteoclast differentiation. Identical insertional mutations in the first exon of RANK have been identified in all published FEO kindreds. The mutation is an 18 base pair tandem duplication in the sequence coding for the signal peptide of RANK, which causes an increase in NF-kappaB signaling. We report the identification and mutational analysis of two unrelated FEO patients. One had no family history of FEO, but presented with bilateral hearing loss at an early age, deterioration of teeth, and severe pain and swelling in the distal tibia before the age of 20. The second patient had a family history of FEO and exhibited an extensive expansile tibial lesion and lesions in one humerus and a phalanx. She also had early hearing loss and dental disease. Mutational analysis of the TNFRSF11A gene in our patients demonstrated an 18 base pair tandem duplication, one base proximal to the duplications previously reported. This novel mutation results in addition of the same six amino acids to the RANK signal peptide that has been observed previously. Further analysis of the exon 1 sequence demonstrated that it has the ability to form a stable secondary structure that may facilitate the generation of tandem duplications. PMID- 12568417 TI - Reduced levels of MMP-2 and TIMP-1 in dyssegmental dysplasia. AB - Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) were measured in a mild case of dyssegmental dysplasia. X-ray pictures of a female baby born vaginally at 39 weeks of gestation showed short, bent, dumbbell-shaped long bones of the limbs and profound dyssegmental ossification in the spine, findings characteristic of dyssegmental dysplasia. When the levels of MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured, the levels of MMP-2 and TIMP-1 were significantly reduced. This case might provide a clue to disclose the etiology of dyssegmental dysplasia. PMID- 12568418 TI - Determinants of bone strength. PMID- 12568419 TI - Development of newer therapies in haemodialysis. PMID- 12568420 TI - Medicine in stamps. Avicenna (980-1037): prince of physicians. PMID- 12568421 TI - Profile of admissions to an acute dialysis care unit. AB - The profile of admissions to an acute dialysis care facility of the National University Hospital Renal Centre, Singapore was studied in an attempt to identify the needs of these patients so that admissions may be possibly reduced. All patients above the age of 18 years who were admitted to the dialysis centre in the National University Hospital for dialysis between 1 Jan 2000 and 31 Mar 2000 inclusive were studied. A total of 124 patients were studied. Almost all the patients had end stage renal failure. The commonest cause of end stage renal failure was diabetic nephropathy (58.8%). There were a total of 157 admissions. Most of the admissions were associated with end stage renal failure (96%). Vascular access problems caused 40% of all admissions and 68% of all multiple admissions. The data suggest that a dialysis facility in a multidisciplinary acute hospital needs to meet the requirements of mainly patients who are already in end stage renal failure, those who have vascular access problems and those who have diabetes. PMID- 12568422 TI - Malignant mixed Mullerian tumours of the uterus--a ten-year experience. AB - OBJECTIVES: To review the clinico-pathological features of malignant mixed Mullerian tumours of the uterine corpus, their prognosis and treatment outcome. METHODS: A retrospective study of malignant mixed Mullerian tumours of the uterus seen at KK Women's & Children's Hospital from January 1989 to December 1998. RESULTS AND CONCLUSION: Twenty-six patients with mean age of 56.5 years were analysed. Twenty (76.9%) were menopausal. None had previous pelvic irradiation. Vaginal bleeding and uterine enlargement were the commonest presenting symptom and sign. Diagnostic dilatation and curettage obtained the diagnosis in 15 patients. Majority of patients had surgery with adjuvant chemotherapy, while adjuvant radiotherapy was offered only recently. Positive peritoneal washings were significantly associated with advanced disease. There were seven patients with stage I, four with stage II, nine with stage III and four with stage IV disease. There were 17 homologous and nine heterologous tumours. Presence of heterologous stromal components did not influence the stage of the disease. Increasing depth of myometrial invasion was associated with poorer survival. Prognosis of patients with stage III and IV disease were poor, with none surviving to two years. All the patients with stage I disease were still alive at the end of the study period. In conclusion, malignant mixed Mullerian tumours of the uterine corpus are aggressive tumours associated with poor prognosis. PMID- 12568424 TI - Medical support for the North East Line Mass Rapid Transit project by the Republic of Singapore Navy's Naval Medicine Hyperbaric Centre. AB - Naval Medicine Hyperbaric Centre (NMHC) was approached by the Land Transport Authority (LTA) to provide medical support for the North Eastern Line MRT Project. The medical support provided by NMHC focused on providing training, audits and ensuring that safety and health infrastructure were in place even before the commencement of compressed air tunnelling. The extensive use of the EPBM (Earth Pressure Balance Machine) tunnelling machines for the first time in Singapore necessitated a reassessment of the medical support system and paradigm that was based primarily on the older form of tunnelling. The survey results showed that human exposure to compressed air works with the use of EPBM was minimal and no decompression illness was reported. However, there was still a total of 28 cases of barotrauma that were reported which were mainly from the tunnels dug using the open face compressed air tunnelling method. As such, medical providers should still exercise careful planning for the medical support of compressed air works. PMID- 12568423 TI - Well-differentiated thyroid carcinoma: factors predicting recurrence and survival. AB - AIMS: 1) Determine the patient and tumour characteristics for well-differentiated thyroid carcinoma--towards developing a unique risk classification for our largely Chinese population. 2) Assess extent of thyroid surgery required. 3) Document prognostic value of UICC and AMES classification. METHODS: Retrospective review of 175 patients treated for primary thyroid epithelial malignancy by the Department of General Surgery at the Singapore General Hospital. RESULTS: There were 78% papillary carcinomas (PC) and 19% follicular carcinomas (FC). Female: male ratio was 3:1. Patient distribution in the UICC stages I, II, III, IV is respectively 56, 11, 31 and 2%. Twenty-six percent had hemithyroidectomy, and 74% total thyroidectomy. Neck dissections were required in 6% of FC compared to 34% of PC. Mean follow-up was 40 months. Extent of surgery did not affect PC/FC survival nor recurrence rates (p=0.53 and 0.06 respectively). Recurrences occurred in 15% FC and 9% PC. Death occurred in one FC and two PC. Survival correlated with UICC stage I/II and stage III/IV groups (p=0.04), and recurrence correlated with AMES High and Low Risk groups (p=0.004). No statistically significant difference was shown for survival between PC and FC or AMES groups and recurrence between PC and FC or UICC groups. CONCLUSIONS: Extent of thyroid surgery does not significantly affect local recurrences of PC/FC. The characterisation of thyroid carcinoma here is an important step towards developing a risk classification unique to our largely Chinese population. PMID- 12568425 TI - Use of percussion as a screening tool in the diagnosis of occult hip fractures. AB - Traumatic hip pain is a common clinical problem in the emergency department. There is significant morbidity in discharging a patient with an undiagnosed undisplaced hip fracture. The auscultatory percussion technique is a useful method to risk stratify patients who present with traumatic hip pain and with normal radiographs. We sought to study the sensitivity and specificity of the auscultatory percussion technique in a prospective study. PMID- 12568426 TI - Premature discharge in a community hospital. AB - AIM: To determine the size of the problem of premature discharge in a community hospital (CH) and to ascertain the reasons for it. METHOD: A retrospective review of all admissions in year 2000 which resulted in premature discharge i.e. discharge within one week of admission, was conducted in a community hospital, St Luke's Hospital for the Elderly. Information collected on the selected cases included biodata, reason for CH stay, admitting diagnosis, source of admission, duration of stay and reason for terminating stay prematurely. For cases which required acute hospital transfer or ended in death in the CH, the type and day of onset of the respective medical problems were documented. RESULTS: Out of 924 admissions in year 2000, 12% resulted in premature discharge. Within this category of patients, 54% were discharged within the first three days and median duration of stay was three days. Majority of the admissions were for rehabilitation (83%) and respite care (15%). Neurological (60%) and orthopaedic (18%) problems constituted the bulk of the admitting diagnoses. The main reason for premature discharge was acute hospital transfer (90%) for medically unstable patients and those with unresolved medical problems. CONCLUSION: Premature discharge in the CH is an important issue and the greater cause lies in the need to transfer medically unstable patients or patients with unresolved medical problems back to the acute hospital. Stricter enforcement of admission criteria into CHs, increased vigilance on the part of acute hospitals and implementation of subacute care in CH can be solutions to the problem. PMID- 12568427 TI - A defective endotracheal tube. AB - Routine inspection and testing of endotracheal tubes prior to use may fail to detect certain manufacturing defects. We describe a case of endotracheal tube kinking at the junction where the inflation tube for the pilot balloon is attached to the endotracheal tube. This case report highlights the importance of maintaining an awareness that airway obstruction or leak due to structural defects can still occur even with high quality, prepacked single use plastic endotracheal tubes. It also emphasises the need to have a systematic approach when dealing with such critical events. PMID- 12568428 TI - Vitamin E and thetreatment and prevention of diabetes: a case for a controlled clinical trial. AB - Strategies to delay the onset and ameliorate the sequelae of type 2 diabetes are urgently needed in Singapore. Diabetes is accompanied by severe oxidative stress (especially lipid peroxidation) due to increased oxygen free radical production. Oxidative stress in part results from hyperglycaemia, but it may also precede, and accelerate the development of overt type 2 diabetes and then of diabetic complications. Epidemiological evidence indicates low vitamin E intake as a risk factor for development of type 2 diabetes, and small scale human intervention studies have indicated benefit of vitamin E in improving endothelial function, retinal blood flow and renal dysfunction. Animal studies also support its usefulness. The weight of evidence available supports the suggestion that a major double-blind controlled clinical trial of antioxidants in prevention and treatment of type 2 diabetes should be undertaken. PMID- 12568429 TI - Clinics in diagnostic imaging (77). Pes anserine bursitis. AB - Many cystic lesions occur around the knee and may produce overlapping clinical features, rendering the clinical diagnosis difficult. A 50-year-old woman presented with a soft tissue swelling on the medial aspect of her right knee. The diagnosis of pes anserine bursitis was made, based on typical MR imaging features. Cystic masses occurring in and around the knee, such as bursae and recesses, meniscal and ganglion cysts, and benign and malignant soft tissue masses that may mimic cysts, are classified and described. The role of MR imaging in making an accurate diagnosis and distinguishing among the various masses is discussed. PMID- 12568430 TI - "Playing through pain". PMID- 12568431 TI - "Virtual" medical schools becoming a reality. PMID- 12568432 TI - Nurses and caring. PMID- 12568433 TI - Attitudes toward care of the terminally ill: an educational intervention. AB - This quasiexperimental study examined the effect of an educational program on attitudes toward caring for terminally ill persons and their families. Participants were 115 undergraduate students: intervention group, N = 49; control group, N = 66. Pre- and post-intervention measurements were done with the Frommelt Attitude Toward Care of the Dying Scale (FATCOD, Form B). Students in the intervention group participated in a semester-long (15-week, 45-hour) educational program. Demographic variables, including age, gender, religion, major area of study, influence of religious beliefs, profession, previous education, and past or present experience with loss were evaluated. Statistical analyses (t-test, ANOVA, ANCOVA, and APVs) indicated a significant positive change in the attitude scores of the intervention group and no significant change in the attitude scores of the control group. PMID- 12568434 TI - Spiritual pain: a comparison of findings from survivors and hospice patients. AB - The article presents comparative research findings on the notion of "spiritual pain." The findings from interviews with hospice patients affirm the previously published, preliminary conceptualization of spiritual pain from interviews with survivors. However, while the survivor findings highlight the potential for spiritual pain associated with life after high-tech curative treatment, the hospice patient data emphasize the protectiveness of the hospice experience for deflecting the possibility of spiritual pain. It is anticipated the discussion of comparative findings will affirm the importance of researching this "ignored dimension" and, in so doing, will enrich our understanding of the spiritual dimension of healthcare. The work is part of a program presently developing a language of spirituality through research. PMID- 12568435 TI - Psychosocial issues in palliative care: physicians' self-perceived role and collaboration with hospital staff. AB - Psychosocial issues are a major part of palliative treatment, yet, due to inadequate training, physicians are often ill-prepared to address them. Twenty physicians were interviewed about the importance they placed on psychosocial issues and the perceptions they had of their role in addressing them. Several respondents felt psychosocial issues were important because they affect physical issues, enable holistic care, enhance relationships, impact care decisions, and can reduce patient and family stress. Other respondents did not feel psychosocial issues were their responsibility due to time constraints, their focus on physical care, their lack of expertise in this area, the patients' preferences for attending physicians, and a sense on the part of house staff physicians of not yet being "real" doctors. Collaboration with other hospital staff helped overcome some of these obstacles. Since physicians must often provide psychosocial care, improved training in addressing psychosocial issues is indicated PMID- 12568436 TI - Hospice and primary care physicians: attitudes, knowledge, and barriers. AB - Underuse of hospice services is a significant problem in the United States. Primary care physicians constitute an increasing referral base and have been hypothesized to be important barriers to increased use. We conducted a mail survey of 131 primary care physicians (overall response rate of 72 percent), examining their attitudes toward, knowledge about, and perceived benefits and barriers to hospice care. Physicians demonstrated very favorable attitudes towards hospice. They had correct knowledge about most aspects of hospice, and, where they did not, they were far more likely to be uncertain than erroneous. Primary care physicians perceived many benefits to hospice care and identified patient and family readiness as the major barrier to earlier hospice referrals. A significant subgroup had concerns about problems in interacting with hospices. There were very few differences between family practitioners and general internists. These findings have many implications for directing collaborative efforts between primary care physicians and hospices to improve end-of-life care. PMID- 12568437 TI - Communication of a cancer diagnosis: patients' perceptions of when they were first told they had cancer. AB - Fifty consecutive cancer patients cared for by a palliative medicine program were interviewed concerning the initial communication of their diagnosis. The majority of patients were satisfied with the manner and the circumstances in which the information was imparted. A minority of women were significantly more unhappy than men about the manner in which they were told. Sophisticated techniques are available to help physicians impart bad news effectively and humanely. PMID- 12568438 TI - Opioids, respiratory function, and dyspnea. AB - Dyspnea, the sensation of difficult breathing, is a common debilitating symptom in advanced cancer and chronic progressive cardiopulmonary disease. Primary treatment is correction of the underlying etiology. In incurable illness wherein the cause is irreversible and the goal is palliation, opioids are the drugs of choice for symptomatic relief. This article reviews current knowledge in the pathophysiology of dyspnea, proposed opioid mechanism of action, and evidence of efficacy. PMID- 12568439 TI - Phenol saddle blocks for intractable pain at end of life: report of four cases and literature review. AB - Four cancer patients with prior bladder diversions had phenol neurolytic saddle blocks performed for intractable pelvi-sacral pain. All patients had advanced disease, the focus of their treatment being palliative. Treatment limiting side effects precluded further upward titration of systemic analgesic therapies. Pain control improved after intrathecal neurolysis and allowed a greater than 60 percent reduction in systemic opiate dosage. No significant block-related adverse effects were encountered. The value and technical aspects of intrathecal saddle blocks in end-of-life pain management is discussed. PMID- 12568440 TI - Cancer statistics, 2003: further decrease in mortality rate, increase in persons living with cancer. PMID- 12568441 TI - Cancer statistics, 2003. AB - Each year, the American Cancer Society estimates the number of new cancer cases and deaths expected in the United States in the current year, and compiles the most recent data on cancer incidence, mortality, and survival by using incidence data from the National Cancer Institute (NCI) and mortality data from the National Center for Health Statistics (NCHS). Incidence and death rates are age adjusted to the 2000 US standard population. In the year 2003, we estimate that 1,334,100 new cases of cancer will be diagnosed, and 556,500 people will die from cancer in the United States. Age-adjusted cancer death rates declined in both males and females in the 1990s, though the magnitude of decline is substantially higher in males than in females. In contrast, incidence rates continued to increase in females while stabilizing in males. African-American males showed the largest decline for mortality. However, African Americans still carry the highest burden of cancer with diagnosis of cancer at a later stage and poorer survival within each stage compared with Whites. In spite of the continued decline in cancer death rates in the most recent time period, the total number of recorded cancer deaths in the United States continues to increase slightly due to the aging and expanding population. PMID- 12568442 TI - American Cancer Society guidelines for the early detection of cancer, 2003. AB - Each January, the American Cancer Society (ACS) publishes a summary of existing recommendations for early cancer detection, including updates, and/or emerging issues that are relevant to screening for cancer. In 2002, the ACS assembled expert groups to update guidelines for cervical cancer screening and breast cancer screening, and to evaluate new technology for colorectal cancer screening. In November 2002, updated guidelines for cervical cancer screening were published in this journal, and breast cancer screening guidelines will be updated in 2003. In this issue, there is a report of a workshop held to review emerging technology for colorectal cancer screening that resulted in a modification of current previous recommendations for fecal occult blood tests, and revised recommendations for the "cancer-related check-up" in which clinical encounters provide case-finding and health-counseling opportunities. Finally, we provide an update of the most recent data pertaining to participation rates in cancer screening by age, gender, and ethnicity from the Centers for Disease Control and Prevention's Behavioral Risk Factor Surveillance System (BRFSS). PMID- 12568443 TI - Emerging technologies in screening for colorectal cancer: CT colonography, immunochemical fecal occult blood tests, and stool screening using molecular markers. AB - The American Cancer Society's (ACS) Colorectal Cancer Advisory Group held a workshop on new technologies for the early detection of colorectal cancer and adenomatous polyps as part of a regular review of ACS guidelines for colorectal cancer screening. The Advisory Group formally reviewed CT colonography, immunochemical fecal occult blood tests (FOBT), and stool screening using molecular markers, and also addressed other technologies including capsule video endoscopy. With the exception of immunochemical stool testing, the ACS has determined that at this time there is insufficient evidence to recommend these technologies for routine colorectal cancer screening. Based on recommendations of the Advisory Group, only a minor modification has been made to the ACS's Recommendations for Screening and Surveillance of the Early Detection of Adenomatous Polyps and Colorectal Cancer. PMID- 12568444 TI - Welcome to the 'bottom line'. PMID- 12568445 TI - Implementing paramedic thrombolysis--an overview. AB - The UK Government has made improvements in cardiac care a high priority. The publication in 2000 of the National Service Framework for Coronary Heart Disease and the NHS Plan set out national standards for the management of suspected heart attack, including challenging targets for reducing treatment delays for administration of thrombolytic therapy. This paper discusses the background, evidence base and challenges of implementing one component of the Government's drive to improve cardiac care: the NHS Plan commitment to a three year programme to equip and train ambulance paramedics to safely provide thrombolysis for appropriate patients. PMID- 12568446 TI - Blunt chest trauma: a challenge to accident and emergency nurses. AB - The incidence of myocardial contusion as a result of blunt chest trauma remains difficult to assess. As such the potentially lethal consequences are often difficult to mitigate against. The true incidence of myocardial contusion is not known and various authors have ascribed percentages in the very broad range of 8 71% of those having suffered blunt chest trauma. The extremely wide variation in presenting signs and symptoms further complicates the clinical picture creating a complicated and complex challenge for the assessing team. In the absence of clear guidelines to date a number of potential options are discussed exploring their efficacy and appropriateness in the management of those patients suffering from blunt chest trauma. PMID- 12568447 TI - The quality of psychiatric services provided by an Australian tertiary hospital emergency department: a client perspective. AB - The mainstreaming of psychiatric services within the general health care system has created fundamental changes to the manner in which clients access acute psychiatric services. A review of the literature suggests that this process has been problematic. The current study involved the conduct of telephone interviews with psychiatric clients (n = 136) to ascertain their level of satisfaction with the services received in the emergency department of a Melbourne Metropolitan Hospital. The results were analysed using descriptive statistics. The study participants indicated a high level of satisfaction. Particular emphasis was placed upon the availability of staff with psychiatric qualifications and experience to provide treatment, support and care. Dissatisfaction was noted by some clients regarding lengthy waiting times, lack of privacy in the triage area and negative attitudes of emergency department staff. These findings support the value of psychiatric consultancy services in the emergency department, and further identify the need for triage guidelines to be tailored to the needs of mental health clients. PMID- 12568448 TI - How can I improve my professional practice? PMID- 12568450 TI - Development of physiological discriminators for the Australasian Triage Scale. AB - The Consistency of Triage in Victoria's Emergency Departments Project (2001), funded by the Victorian Department of Human Services, aimed to improve the consistency of application of the Australasian (National) Triage Scale (ATS). One of the major objectives of the project was the development of an education strategy to promote a consistent approach to triage education, leading to the development of the Adult Physiological Discriminators (APDs) for the ATS and Paediatric Physiological Discriminators (PPDs) for the ATS. The guidelines and physiological discriminators were developed in consultation with the Emergency Nurses' Association of Victoria (ENA Vic.) and clinical nurse educators, lecturers, nurse unit managers and clinicians from a wide variety of Emergency Departments (EDs) across Victoria. Numerous studies have identified varying degrees of inconsistency in the application of the ATS. A number of factors associated with inconsistency in the application of the ATS have also been alluded to in the literature. These range from the wide variation in the experiential and educational requirements of Victorian triage nurses to the specific clinical characteristics of the patient identified by the triage nurse. However, a consistent approach to triage education and uniform triage guidelines has been repeatedly identified as a key factor in improving the consistency of application of the ATS. Physiological data demonstrates the highest degree of objectivity and consistency and research has shown that physiological observations are useful and measurable indicators of clinical urgency and patient safety. This paper will discuss the development of these discriminators as part of the educational strategy including a critique of other approaches to triage decision-making and a review of the consultative processes used to facilitate consensus amongst triage nurses, ED Nurse Managers and ED Nurse Educators. The physiological discriminators developed by this project are also presented. PMID- 12568449 TI - Recommendations for nursing requirements at a field hospital, based on the Israel Defense Forces field hospital at the earthquake disaster in Turkey--August 1999. AB - On the 17th of August 1999, an earthquake of 7.4 magnitude on the Richter Scale struck the Marmara region in Turkey causing a massive casualties event with an estimated 2,680 deaths and 5,300 injuries just at the city of Adapazari alone. A field hospital was set up by the Israel Defense Forces at Adapazari in order to provide temporary medical services until regular medical forces recovered. The aim of the paper is to overview the requirements of the nursing staff at a field hospital based on our experience and analysis of the nursing activity at the field hospital at Adapazari. The methods implemented include interviewing all nurses and many of the doctors who took part in the field hospital as well as a review of medical literature about disasters. We found an inverted nurse:phycisian ratio of 1:1.77, as opposed to a 2.5-3:1 ratio in regular civilian hospitals. The nurses in our field hospital had to work longer and more intensive shifts than in a regular hospital. They had to overcome language barriers and cultural differences, and faced difficult hygiene conditions. Our overview analysis of results brought up several recommendations. Firstly, although it is not possible to predictthe number and types of casualties, it is necessary to provide an adequate number of nurses (1-1.5:1 nurse:physician ratio). Furthermore, the nurses should be specialized and rotated as needed. Secondly, the language and cultural barriers should not be undermined despite the abundance of translators. Finally, the hygiene status in a field hospital requires management by nurses with active participation of all members. PMID- 12568451 TI - Emergency care of the crucifixion victim. AB - Of all the terrible ways to die, most people say that death by fire or death by drowning are the worst. If you lived 2000 years ago however, you most certainly would disagree. Throughout world history, one of the most feared deaths was that of crucifixion. This article will guide you through the medical, psychological, and emotional aspects of crucifixion. The death of the man called Jesus Christ will be used to illustrate the use of a punishment that was unequalled in its cruelty and depth of suffering. This article will review not only the injuries associated with crucifixion, as well as current medical archaeological theories relating to the suffering and eventual death on a cross, but also using the introduction case study, the initial assessment and resuscitation of a crucifixion victim will be addressed. Regardless of religious beliefs, this article will give attendees a deeper awareness of "and they crucified him". PMID- 12568452 TI - Effects of pesticides on DNA and protein of shrimp larvae Litopenaeus stylirostris of the California Gulf. AB - Recently, diverse pathologies and massive mortalities have been presented in shrimp hatcheries located along the California Gulf; therefore, toxic responses of shrimp larvae were used as biomarkers of pesticide pollution, because in this region intensive agriculture is practiced. Shrimp larvae were exposed to DDT, azinphosmethyl, permethrine, parathion, chlorpyrifos, malathion, endosulfan, and carbaryl, in order to determine LC50, DNA adducts and/or breaks, and total protein in larvae. The results indicate reductions in protein and DNA in larvae exposed to these pesticides, and in those exposed to DDT, breaks and/or adducts were registered. It is possible that pesticide pollution is a cause of these problems, because reduction in protein indicates a decrease in larvae growth rate and DNA breaks or adducts have been related to pathologies and carcinogenesis in many aquatic organisms. PMID- 12568453 TI - Photonegative response in brown planaria (Dugesia tigrina) following regeneration. AB - The photonegative response was investigated in regenerated brown planaria (Dugesia tigrina). Old, middle, and young animals were tested, bisected into heads and tails, and allowed to regenerate. Different regeneration times affected function, with generally older planaria achieving full functionality of the photonegative response before younger planaria. Counterintuitively, the heads of the middle and young lost this function initially (despite only needing to regenerate their tails) and regained it over time. The size of the animal was not a factor in the photonegativity response. The biological basis for the photonegative response is complex, requiring nervous and locomotive system function and integration. Using the heads from young planaria may serve as a model in aging, degenerative diseases, or environmental toxins. PMID- 12568454 TI - Nutrition influence on copper accumulation by Brassica pekinensis Rupr. AB - Effect of nutrition status in the growth medium on Cu accumulation and plant growth were tested in Brassica pekinensis grown in hydroponic culture in a cultural facility. Cu concentration in the roots and shoots increased with increasing external Cu in the culture solution. On the other hand, it decreased with increasing Hoagland's solution (HS) strength. Interaction between external Cu and HS strength was also observed to affect Cu concentration in the tissues. A low level of external Cu could lead to a high Cu concentration in the plant at low HS strength, while to obtain a high Cu concentration in the plant at high HS strength, a high level of external Cu was needed. Partial nutrient experiments showed an important role of Fe and P in governing Cu accumulation. The influence of nutrients on Cu accumulation were mainly attributed to the osmolarity of the nutrient solution, antagonism between nutrients and Cu, pH change, and cation transporter induction. Root length and shoot biomass were negatively related to Cu concentration in the plant, suggesting an adverse effect of Cu in the tissue on plant growth. Root biomass showed less response to Cu concentration in the plant. This was presumably due to plant resource allocation between shoot and root, by which more resources were shifted to the Cu-stressed roots. The results might have important implications in understanding the sterility of the growth media of plants on Cu fate and effect in the ecosystem. PMID- 12568455 TI - Effect of fenvalerate-20EC on sericigenous insects. I. Food utilization in the late-age larva of the silkworm, Bombyx mori L. AB - The effect of the common pyrethroid insecticide fenvalerate-20EC (Sumicidine 20EC) on food utilization parameters in the pure multivoltine (Pure Mysore) and pure bivoltine (NB4D2) races of silkworm, Bombyx mori, fed ad libitum with Morus alba, was studied in the final two instars. Insecticide treatment was found to reduce the rates of feeding, assimilation, and conversion along with the efficiencies of conversion of ingested and digested food into body substance. This was due to the toxic effects, which interfered with the silkworms' feeding behavior, and physiology of digestion, and the magnitude of changes increased with the increase in the concentration of fenvalerate used. To overcome the deleterious effects of such pyrethroid insecticides, it has been suggested that the safe periods for use of mulberry leaves contaminated with fenvalerate for summer and winter. PMID- 12568456 TI - Effect of fenvalerate-20EC on sericigenous insects. II. Digestive enzymes in the nutritive physiology of silkworm, Bombyx mori L. AB - Changes in the activities of a few digestive enzymes after fenvalerate-20EC treatment were studied during the late stages of multivoltine and bivoltine silkworms, Bombyx mori. Insecticide treatment induced a reduction in the activity of such enzymes as amylase, sucrase, and protease, whereas the trehalase activity was enhanced when the midgut was used as the source of enzyme. The changes recorded in the activity of these enzymes indicated probable adverse effects of fenvalerate on silkworm metabolism through interference with the functional abilities of digestive enzymes. PMID- 12568457 TI - Bioluminescence assays: effects of quinones and phenols. AB - The influence of a series of quinones and phenols on bacteria bioluminescence systems was investigated. Three bioluminescence systems used in ecological monitoring were compared: (1) water-soluble; (2) immobilized in starch gel coupled enzyme systems: NADH:FMN-oxidoreductase-luciferase; (3) luminescent bacteria. Bioluminescence inhibition constants of quinones and phenols and bioluminescence induction periods were compared. These kinetic parameters are proportional to quinone concentrations and depend on the quinone redox potential. Different effects of the substances are related to structure and properties of the bioluminescence systems. The set of bioluminescence assays for quinones and phenols monitoring should include two bioluminescence systems: 1 (or 2) and 3. PMID- 12568458 TI - Confounding factors in bioassays with freshwater and marine organisms. AB - The use of bioassays in ecological risk assessments often raises questions about the causative factors, and insight into the possibility that confounding factors, such as pH or increased ammonia concentrations, might be responsible for the observed toxicity is needed. It was decided to develop a practical approach for the Dutch situation, in which a first screening is carried out based on provisional criteria. In collecting the required data, dozens of experiments were performed, while the scientific literature was searched for additional information. It is concluded that the provisional criteria specified are at present useful tools in interpreting results of bioassays. Depending on the outcome and the aim of the research, it might be necessary to further reduce uncertainties in the interpretation. This might require some additional experiments, using alternative controls or test procedures or altering the composition of the original sample. PMID- 12568459 TI - Biomarker responses and resin acids in fish chronically exposed to effluents from a total chlorine-free pulp mill during regular production. AB - This study used rainbow trout (Oncorhynchus mykiss) to investigate the biological effects of long-term exposure to a total chlorine-free (TCF) pulp mill effluent. Fish tanks were set up on the premises of the pulp mill, and fresh effluent water was led through a pipe directly from the pulp mill to the tanks. The fish were exposed to effluent for up to 50 days and kept for up to 70 days afterward to study the recovery process. Two independent experiments were carried out. Ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), and glutathione reductase (GR) activity were elevated in fish exposed to a 2% concentration of pulp mill effluent, as were levels of DNA adducts. These effects could be detected during exposure and also after a period of recovery. The results demonstrate that the pulp mill effluent contains substances that affect the detoxification process and also have genotoxic potential. The continued occurrence of effects after a 70-day recovery period was demonstrated in both experiments, and may indicate that the effluent contained compounds with persistent properties. The content of free and conjugated resin acids in the bile of the fish was found to be a useful indicator of exposure to pulp mill effluent. PMID- 12568460 TI - Low concentrations of lorsban in water result in far reaching behavioral and histological effects in early life stages in guppy. AB - Chlorpyrifos (O,O-diethyl-O (3,5,6-trichlor-2-pyridyl) phosphorothioate) is a broad-spectrum organophosphate compound. A number of previous work of different countries, institutes, and individuals have confirmed that chlorpyrifos is very highly toxic to freshwater fish and to aquatic invertebrates. The acute toxicity and subacute toxic effects on different concentrations of chlorpyrifos in guppy were investigated in the current study. Twelve guppy juveniles (x 6) were exposed to a range of chlorpyrifos concentrations and 96 h. There was one control group. Subacute exposures were predetermined based on 96-h LC50 values. Fish were exposed to three concentrations 0.5, 1, and 2.0 microg/L chlorpyrifos (Lorsban) for 14 consecutive days. Numbers of live and dead juveniles, signs of paralysis, and deformations were monitored every 24 h. The acute toxicity results revealed that even low concentrations of Lorsban were toxic to guppy juveniles, and with toxicity being evident from 7.17 microg/L LC50 for 96 h at 26 +/- 1 degree C. Low concentrations of Lorsban were also responsible for changes in swimming behavior. Malformations were common to all treated groups but increased in the highest exposure group. Gill histology revealed several pathological changes, including loss and shortening of secondary lamellae. Collectively, the present study reveals that low concentrations of chlorpyrifos can be highly detrimental to guppy in the early life stages, with clear evidence of behavioral, morphologic, and histologic effects. The present findings warrant further studies. PMID- 12568461 TI - Toxicity study of diethyl phthalate on freshwater fish Cirrhina mrigala. AB - Diethyl phthalate (DEP) is used as a plasticizer, a detergent base, in aerosol sprays, as a perfume binder in incense sticks and after-shave lotions. It is known to be a contaminant of freshwater and marine ecosystems. Therefore, a study was designed to determine the toxic effects of DEP on a freshwater fish, Cirrhina mrigala. The fish was treated with 25, 50, 75, and 100 ppm (w/v) DEP dissolved in acetone to determine the LC50. Positive controls were treated with acetone only. There was 100% mortality observed within 24 h in 75 and 100 ppm, and 50% mortality in 50 ppm treated fish in 72 h. Those treated at 25 ppm showed only 10% mortality within 72 h and remaining fish continued to survive. The surviving fish were treated with 25 ppm DEP once daily for 3 days with every change of water (Group III). One group was maintained as negative control in dechlorinated water (Group I) and the other group received acetone once daily for 3 days with every change of water and was used as positive control (Group II). Fish were killed by cold narcosis on an ice block and dissected to obtain liver, muscle, and brain samples; 10% homogenates in ice-cold saline were prepared. Brain and muscle acetylcholinesterase (AchE) activity was measured. Liver aspartate (AST) and alanine aminotransferase (ALT), and liver and muscle succinate dehydrogenase (SDH) alkaline and acid phosphate (ALP and ACP) were measured. There was a significant increase in liver and muscle ACP and ALP in DEP-treated fish compared with positive and negative controls. There was a significant increase in muscle SDH and liver ALT (ALT) in DEP-treated fish compared with positive and negative controls. Brain AchE level was significantly decreased in DEP-treated fish compared to positive and negative controls. These results indicate that DEP brings about significant changes in the activity of certain liver and muscle enzymes. These alterations in enzyme activity may have long-term effects on that are continuously exposed to low doses of DEP in the aquatic environment. PMID- 12568462 TI - Lead intoxication in gestational and lactation periods alters the development of male reproductive organs. AB - The effect of lead (Pb) intoxication during pregnancy and lactation on the male reproductive system was studied to evaluate the alterations caused by Pb in the development of pups. The investigations covered the effect of lead on the course of spermatogenesis and the development of the epididymis and reproductive glands. For this purpose, dams were intoxicated with 300 mg/L Pb during the gestational period and through lactation. Pups were sacrificed on Postnatal (PN) Days 12 and 21. Blood lead (PbB) and plasma iron concentrations were measured, and blood cells counted. Biochemical studies as well as histochemical analyses were performed on testes and accessory glands of the reproductive system. Lead intoxication resulted in a decrease in testis and seminal vesicle weights and an increase in DNA and RNA levels on PN Day 21. Total protein was significantly decreased by the toxicant, and alkaline and acid phosphatase levels of the gonads were reduced. Effects were also reflected in the reduction of the thickness of epithelium and of seminiferous tubule diameter (STD) as a consequence of the action of lead in the reduction in numbers of prospermatogonia and spermatocytes. Results indicate that the reproductive system targets of lead intoxication are not only the testes; lead intoxication results in the inhibition of testicular, epididymal, and seminal vesicle function, altering the biochemical composition of these organs, and consequently, affecting the normal development of germinal cells. PMID- 12568463 TI - Effects of Sevin (carbaryl insecticide) on early life stages of zebrafish (Danio rerio). AB - Sevin brand carbaryl insecticide is one of the most commonly used insecticides in the United States, with great potential for leaching into ground- and surface water reserves. Its possible teratogenic effects were tested on zebrafish eggs in four dilutions of decreasing concentration. The average mortality rate was low, indicating that Sevin does not directly kill embryos at these concentrations. Eggs and embryos were consistently smaller than the control starting at 24 h after spawning until hatching. Embryos in the highest concentration took up to twice as long to hatch as the control. This delayed hatching time increases vulnerability to predation. In addition, as minnows are lower on the food chain, bioaccumulation of Sevin in tissues may increase in larger predators, affecting their metabolism and reproduction. PMID- 12568464 TI - Acute toxicity of drainage ditch water from a Washington State cranberry-growing region to Daphnia pulex in laboratory bioassays. AB - High concentrations of organophosphorous insecticides resulting from cranberry bog applications were detected in the Grayland Drainage Ditch (GDD) system in Grayland, Washington State, during the 1994-1996 Washington State Department of Ecology Pesticide Monitoring Program. This drainage ditch system drains cranberry bogs and enters the Pacific Ocean via the North Cove and Supon Inlet. Concerns about the impact of these pesticides on human and environmental health led to this investigation of the potential impact on an indicator species, Daphnia pulex. To determine the toxic effects of multiple pesticides entering the GDD, standardized laboratory toxicity tests with D. pulex were conducted concurrently with the Washington State Department of Ecology pesticide sampling. Concentrations of three insecticides, diazinon, chlorpyrifos, and azinphosmethyl, were the highest ever detected in state waters. The GDD water was found to cause acute toxicity in 33% of the laboratory bioassays conducted. Regression analysis, however, detected a poor correlation between total insecticide detected and percentage mortality of D. pulex at the two drainage ditch sites studied, Grays Harbor County site and the Pacific County site. However, the relationship between mortality of D. pulex and detected concentrations of diazinon and chlorpyrifos were significant. Sampling schedules for chemical analysis and bioassay testing appear to be the primary reason that statistical analysis failed to correlate mortality with detected OP pesticide concentrations. Grab samples used in toxicity testing may over- or underestimate actual concentrations of contaminants present in the system being studied. PMID- 12568465 TI - Effect of lead and ancylostomiasis on phosphatases of heart in Swiss albino mice. AB - Administration of 0.01 and 0.1 mg of lead nitrate for 4 and 7 days and infection of Ancylostoma caninum larvae orally altered the activation of alkaline phosphatase and acid phosphatase in the hearts of mice when compared to infected animals and controls. Alkaline phosphatase activity increased significantly in all drug-treated + infected mice. The level of acid phosphatase decreased significantly in mice exposed to chronic doses of lead. The altered levels of alkaline phosphatase and acid phosphatase suggest that administration of lead could cause toxicity in the heart, disturbing the cellular metabolism; infection alone could not cause any significant changes in enzymes of heart. PMID- 12568466 TI - Glutathione and metallothionein status in an acute response by Mercenaria mercenaria brown cells to copper in vivo. AB - Red glands of Mercenaria mercenaria comprise brown cells that accumulate, detoxify, and excrete copper. Brown cell involvement in metal detoxification is due in part to endogenous glutathione (GSH) and metallothionein (MT). The intent of this study was to test the hypotheses that brown cell GSH functions in protection against Cu2+ toxicity, that brown cell GSH provides the initial defense against Cu2+ prior to brown cell MT induction, and that MT variants (MTI, MTII), if present are unequal in response to Cu2+. Brown cells were analyzed for GSH and MT after 0.25, 1, 2, 3, and 4 days of treatment of Mercenaria with 0.01 and 0.05 ppm Cu2+. Glutathione initiated the brown cell acute response (within the first day of treatment) to both the 0.01 and 0.05 ppm Cu2+ treatments. Metallothionein in brown cells increased to Day 4 during treatment with 0.01 ppm Cu2+, whereas MT concentration was greatest at Day 2 after which it decreased to Day 4 with treatment of 0.05 ppm Cu2+. The change in MTII relative to its control was greater than that of MTI in the brown cell acute response to 0.01 ppm Cu2+ and also for Days 0.25 to 2 in response to the 0.05 ppm Cu2+ treatment. At Days 3 and 4 with the 0.05 ppm Cu2+ the change in MTI/MTII ratio was due to a greater change in MTI than MTII relative to their respective controls. The variants of brown cell MT appear to respond differently to Cu2+ depending on the Cu2+ treatment concentration. PMID- 12568467 TI - Cytological and biochemical effects of a mixture of 20 pollutants on isolated rainbow trout (Oncorhynchus mykiss) hepatocytes. AB - Isolated hepatocytes of rainbow trout (Oncorhynchus mykiss) were exposed to different concentrations (1x, 10x, 50x) of a complex mixture of 20 environmentally relevant contaminants (PAHs, PCBs, pesticides) typical of the chemical burden of surface waters and sediments of small rivers in southwest Germany to investigate sublethal cytological and biochemical alterations. Results document that all concentrations clearly induced biochemical and morphological changes. The activities of enzymes such as lactate dehydrogenase and catalase as well as the rate of lipid peroxidation were significantly increased, whereas acid phosphatase activity decreased. Cytopathological effects included deformation of cellular shape, deformation and dilation of the nuclear envelope, increase in heterochromatin, nuclear inclusions, heterogeneity of mitochondria, vesiculation of RER cisternae, as well as increases in myelinated bodies, lysosomes, and peroxisomes. If compared with cytological alterations induced in isolated rainbow trout hepatocytes after exposure to sediment extracts from rivers of corresponding contamination, cytological effects of exposure to the complex chemical mixture displayed numerous similarities. PMID- 12568468 TI - Bioaccumulation and detoxication of nodularin in tissues of flounder (Platichthys flesus), mussels (Mytilus edulis, Dreissena polymorpha), and clams (Macoma balthica) from the northern Baltic Sea. AB - Cyanobacterial hepatotoxin accumulation in mussels (Mytilus edulis, Dreissena polymorpha), clam (Macoma balthica), and flounder (Platichthys flesus) tissues was measured. Flounder were caught with gillnets from the western Gulf of Finland on 21 August 1999, 25 July 2000, and 25 August 2000. Blue mussels were collected from: (1) a steel cage at a depth of 3 m on 20 August 1999, (2) an enclosure at depths of 3-5 m, and (3) an artificial reef (wreck at 25-30 m) in the western Gulf of Finland between June and September 2000. Furthermore, blue mussels were collected from two sites between August and October 2000: south of the town of Hanko at depths of 5 and 20 m in the western Gulf of Finland and south of the city of Helsinki at a depth of 7 m in the central Gulf of Finland. M. balthica and D. polymorpha were collected at a depth of 12 m from Russian waters in the eastern Gulf of Finland on 1-4 August 2000. The samples were analyzed for the cyanobacterial hepatotoxins nodularin (NODLN) and microcystins (MCs) using enzyme linked immunosorbent assay (ELISA), liquid chromatography-mass spectrometry (LC MS), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). ELISA indicated a time-dependent accumulation of hepatotoxins in flounder liver up to 400 +/- 10 (SD) microg/kg on 25 August 2000. No hepatotoxins were detected in flounder muscle samples. In blue mussels, collected from an enclosure 3-5 m deep in the western Gulf of Finland on 23 August 2000, ELISA indicated cyanobacterial hepatotoxins up to 1490 +/- 60 microg/kg dry wt. Blue mussels collected from the other sites contained less cyanobacterial hepatotoxins (40-130 microg/kg dry wt). Clams and mussels from Russian waters contained cyanobacterial hepatotoxin at about 100-130 microg/kg dry wt. Total hepatotoxin levels in mussels from enclosures decreased from August to September, indicating at least partial detoxication/depuration of the toxins. LC-MS verified the presence of NODLN in mussels and flounder. Typical detoxication conjugates were observed by MALDI-TOF-MS in mussel samples collected during August 2000. In deeper-living wreck mussels cyanobacterial hepatotoxin levels continued to increase, from August to September, indicating that portions of cyanobacterial hepatotoxins reach the sea floor. NODLN bioaccumulation is a constant phenomenon in the area. PMID- 12568469 TI - Physiological responses of Carassius auratus to ytterbium exposure. AB - Physiological and biochemical perturbations in the liver of Carassius auratus were investigated in vivo following 40 days of exposure to ytterbium solutions of different concentration. Glutamate-pyruvate transaminase (GPT) activity in goldfish liver was stimulated at 0.05 mg/L Yb3+ and inhibited at higher Yb3+ concentrations. Activity of the antioxidant enzyme superoxide dismutase (SOD) was stimulated at Yb3+ higher than 0.05 mg/L, and catalase (CAT) activity was strongly inhibited after 40 days of exposure. Detoxifying enzymes glutathione S transferase (GST) and glutathione peroxidase (GSH-Px) were stimulated at 0.05 mg/L and inhibited at 0.1 mg/L after 40 days of exposure. Among the parameters determined, CAT in goldfish liver was most sensitive to Yb3+, indicating that CAT might be considered a potential tool in the biomonitoring of exposure to Yb3+ in an aquatic ecosystem. PMID- 12568470 TI - Uptake of copper complexed to EDTA, diaminoethane, oxalic acid, or tartaric acid by neon tetras (Paracheirodon innesi). AB - Uptake of copper complexes by fish was studied by exposing neon tetras (Paracheirodon innesi) to solutions containing different concentrations of copper complexed to EDTA, diaminoethane, oxalic acid, or tartaric acid, while other copper species were kept at low and constant levels. The distribution of the copper species in the solution was quantified using chemical equilibrium modeling. It was found that the presence of the organic ligands studied could reduce, but not totally eliminate, the uptake of copper by fish. The rate of the additional uptake besides the contribution of free copper depended on the conditional stability constant of the copper complexes. PMID- 12568471 TI - Microbial toxicity tests and chemical analysis as monitoring parameters at composting of creosote-contaminated soil. AB - Traditionally, chemical analyses are used in the assessment of contaminated soil and in monitoring the efficiency of soil remediation processes. We investigated if chemical analysis could be supported and even partly replaced by biological toxicity tests. In two case studies creosote-contaminated soil was composted outdoors in 5- and 100-m3 windrows. Degradation of polyaromatic hydrocarbons (PAHs) was followed by chemical analysis and toxicity tests. Polyaromatic hydrocarbons were quantified and identified by HPLC. Because the soil was also contaminated by copper-, chromium-, and arsenic-containing fungicides, these elements were analyzed by atomic absorption spectrometry. The toxicity of soil samples was assessed by a soil-contact modification of the luminescent bacteria (Vibrio fischeri) test and in the other case also by enzyme synthesis inhibition (Toxi-ChromoPad test, Escherichia coli). The toxicity of soil water extracts was measured by the standard luminescent bacteria (V. fischeri) test and bacterial (Pseudomonas putida) growth inhibition test. After the first 4 months of the composting period the total amount of PAHs was reduced in all windrows, and in particular, the loss of two- and some three-ring compounds was high, almost 90%. Toxicity decreased concurrently with the decrease in PAH concentration during composting, but after 4 months, one of the piles inoculated with mycobacteria and containing more three- and four-ring compounds was found to be more toxic than at the beginning. After the next summer, total PAH content was further reduced but some four-ring or heavier compounds were demonstrated to be poorly degraded. The toxicity was also reduced to the same level as in the control pile. The total PAH content and the toxicity were both reduced significantly during 5 months of composting. PMID- 12568472 TI - Safe communities for children: only models of good practice or ways through practical difficulties in injury prevention? PMID- 12568473 TI - Parents as experts: the position of parents of children with profound multiple disabilities. AB - BACKGROUND: Parents of children with disabilities are increasingly considered as experts in the field of care. Their expertise can deliver an important contribution towards planning their child's care and education. The law is increasingly taking this factor into consideration. On the one hand, parents already possess an abundance of know-how; on the other, they still have to increase their knowledge and sometimes their skills too. AIM: The purpose of this study is to demonstrate the supposition that co-operation between parents and professionals must meet certain criteria if parents are to receive a proper chance of using their existing knowledge, while at the same time adding to their skills. DESIGN: A questionnaire was sent to 723 parents of children with profound multiple disabilities enquiring about their relationship with the professionals at their child's school for special education. Subsequently, for one group of parents, a method that recognizes the parents' expertise and assigns them a formal role was introduced. For the other group, no change in method occurred. RESULTS: The results demonstrate that parents regard co-operation in a favourable light. Comparing both groups after a year, the expected effect of the method (a better rating of the co-operation by the parents) could not be demonstrated. This is probably a result of a problem with the questionnaire used in this study. CONCLUSION: Other studies, however, appear to support our supposition. PMID- 12568474 TI - Health of children considered for adoption. AB - Children in the 'care' system are known to have poor health. Although health services can respond to and deal with health problems, it is parents, foster parents and social workers who recognize problems, seek advice and keep follow-up appointments. Many health needs remain unmet in such children. This short paper from a deprived area in north-east England shows that once a decision is made to proceed to adoption, the stable and consistent care that can then be provided meets the increased health needs that such children have. PMID- 12568475 TI - Parental participation in the habilitation process--evaluation from a user perspective. AB - AIM: To develop a national instrument for evaluation of parental participation: (1) to obtain a functional measure of quality from a user perspective; (2) as part of quality development in child habilitation services departments; (3) to create common grounds for the evaluation of important aspects of the habilitation process based on the opinions of users and care professionals; (4) to enable evaluation of individual service departments from a more general viewpoint and to highlight areas for improvement; and (5) to enable comparisons of individual service departments in the future against those of others via benchmarking. METHOD: The Measurement of Processes of Care (MPOC) was deemed to be the method that corresponded most closely with these formulated aims. A shortened version, MPOC 20, had already been produced and was awaiting publication. This shortened version measures the same important aspects of habilitation as the original MPOC. It also has a new scale, with verbal clarification for each step. This makes it more user friendly, as the results are easier to interpret. MPOC 20 was modified to become MPOC 28. This questionnaire was sent out in 11 of 26 counties in Sweden. The target group for the questionnaires was families with children up to 18 years of age who had been in contact with a habilitation services department for at least 1 year. The sample group comprised 4013 randomly selected families. A total of 3391 (84.5%) returned the questionnaire, and 2458 (61%) had responded to the questions. RESULTS: Twelve particular questions that can be regarded as fundamental to the habilitation processes emerged from the questionnaire in the regression analysis. These are measures of good quality in the habilitation process as perceived by the parents and are important in their overall satisfaction with habilitation services. Apart from the specific information category, these questions represented all the factors, i.e. enabling/partnership, general information, co-ordinated/comprehensive care and respectful/supportive care. CONCLUSION: MPOC 28 can be useful as an analytical tool for comparisons over time and for measuring changes in the way in which parents rank the various question areas linked to their overall level of satisfaction with the habilitation services in general. PMID- 12568476 TI - Use of physiotherapy and alternatives by children with cerebral palsy: a population study. AB - OBJECTIVES: To describe the use of physiotherapy services and alternative therapies by a population of children with moderate to severe cerebral palsy (CP). DESIGN: Descriptive cross-sectional survey. SUBJECTS: A total of 212 parents of children aged 4-14 years with moderate to severe CP were identified from the Northern Ireland Cerebral Palsy Register (NICPR) and a random subsample of their paediatric physiotherapists. MAIN MEASURES: A standardized description of motor impairment or assessment form; a postal questionnaire to parents and paediatric physiotherapists (to validate parents' reports of service use). RESPONSE RATES: In total, 85% of parent questionnaires were returned and 100% of paediatric physiotherapists responded. RESULTS: Service use among families was high; on average the families had contact with approximately seven services in a 6-month time interval. The overwhelming majority of children (96%) received physiotherapy during the school term and most (59%) received treatment at least twice a week for 30 min; 43% of children had their physiotherapy discontinued over the summer holidays. Over one-quarter (28%) of families had opted out of the NHS and bought alternatives like conductive education (21%) or private forms of conventional physiotherapy (16%). Children with more severe forms of CP, in special education, particularly at schools for physical disability, were high intensity users of the physiotherapy service. Despite this, 74% of parents wanted more physiotherapy for their child. CONCLUSIONS AND IMPLICATIONS: The demand for physiotherapy services is likely to continue given the relatively stable prevalence rate of CP, the proportion of children with disabling CP and the level of parent interest in the service. A number of quality aspects and gaps in the service have been identified. PMID- 12568477 TI - The meaning of parental illness to children: the case of inflammatory bowel disease. AB - BACKGROUND: Research on the impact of parental illness on children is sparse, and it remains unclear what kind of support, if any, families would value. This paper presents findings from the first investigation to explore the experiences of young people who have a parent with inflammatory bowel disease (IBD). DESIGN AND METHODS: This qualitative study involved interviews with 23 young people, aged between six and 20 years, who have a parent with IBD. A range of non-verbal techniques was used to facilitate interviews with young children. Data were analysed using the framework approach. RESULTS: Participants varied in their understanding and perception of their parent's condition and the extent to which they discussed it with family and friends. Some reported being unaffected by their parent's illness. The main negative effects on everyday life were restrictions on social activities, parents being unable to do housework, children having to be well-behaved and parents becoming withdrawn or irritable. Experiences could lead to feelings of sadness, worry, anger and guilt. The few who reported positive effects described spending time with the parent while ill, being allowed to be more independent and being close as a family. Relatives, friends, neighbours, service providers and young people all played a role in helping to manage the situation, and most participants felt that their family did not need additional support. However, some young people would value advice and information regarding the condition and opportunities to meet others in a similar situation. CONCLUSION: IBD is a highly variable condition, with substantial differences in the extent to which it impinges on the lives of family members. In addition to supporting patients, health professionals have an important role to play in offering advice and information to others affected by the illness, including children. PMID- 12568478 TI - A prospective study of absence for illness and injury in childcare children. AB - OBJECTIVES: This population-based study investigated prospectively collected absence episodes to describe patterns and rates of absences for illness and other reasons in two types of formal childcare. DESIGN: Centre day care and family day care homes were selected at random from licensing lists. All centres (n = 11) and 95% of family day care caregivers (n = 130) agreed to participate. The results are based on absence information from carers and on questionnaires to the parents of participating children (n = 846). RESULTS: The major reason for absence was respiratory illness with 48% of all children absent at least once during the year with some form of respiratory illness. Although the absence rate for children in centre care was higher than that in family care (3.34 episodes per 52 child-weeks compared with 2.41), the difference was not statistically significant. CONCLUSION: Investigating patterns of absences from childcare allows a better understanding of needs in childcare. The major reason for absence for illness was respiratory illness. PMID- 12568479 TI - Oscillating between hope and despair--a qualitative study. AB - OBJECTIVE: The aim of this qualitative study was to elucidate the life situation and psychosocial processes of living with chronic pain in children suffering from juvenile chronic arthritis (JCA). METHODS: Taped open qualitative interviews with 22 children (aged 6-17 years) were transcribed verbatim and analysed using the comparative method for grounded theory. RESULTS: A core category, labelled oscillating between hope and despair, was identified and related to four additional categories labelled disturbed order, dependency, ambivalence and uncertainty about the future. There were relationships between the children's subjective experience of pain and their experiences of disturbed order in daily life, dependency on treatment, health care and significant others, ambivalence related to environmental reactions and uncertainty about the future. Chronic pain and disease control the children's lives and lead to restricted participation in social life. CONCLUSIONS: Chronic pain is a substantial problem for children with JCA, which must be considered in health care. The pain affected and disturbed the children's ordinary way of life. It blocked a number of goals. Psychological and physiological processes interact in the phenomenological experience of pain, and the response of the environment elicits, maintains or decreases the experience of pain. Hopefully,the findings contribute to a deeper understanding of the life situation of children suffering from JCA and can be of importance in developing rehabilitation programmes for these children. PMID- 12568480 TI - Omission of active commuting to school and the prevalence of children's health related physical activity levels: the Russian Longitudinal Monitoring Study. AB - BACKGROUND: Active commuting to school by walking or bicycle is a potential source of continuous moderate activity for children that has been largely ignored in surveys of physical activity. The purpose of this study was to evaluate the analytical impact of omitting active commuting to school (walking or bicycling) on conclusions about children's physical activity levels. METHODS: The Russian Longitudinal Monitoring Study (RLMS) is the first nationally representative household survey in the Russian Federation. More than 6400 households from all regions of Russia were surveyed eight times between 1992 and 1998. Analysis was conducted using physical activity data (school physical education classes, out-of school active pursuits and active commuting to school) obtained by parent-proxy on 1094 (572 boys, 522 girls) school-aged Russian children (mean age 10.2 +/- 1.9 years) participating in the November 1998 round of the RLMS. Data were examined according to prevalence of achievement of health-related physical activity guidelines, active commuting to school behaviours included then omitted. RESULTS: Omitting active commuting to school resulted in a statistically significant decrease in the prevalence of achievement of health-related guidelines from 12% to 20%, similar for both genders. Likewise, the prevalence of sedentarism (defined as not meeting any of the guidelines) was increased by 17-22%. CONCLUSIONS: The present findings suggest that, in order to avoid misclassification bias of children's physical activity levels, it is necessary to include questions about mode of commuting to school. The findings also carry practice implications: the commonplace need to get to and from school may be a missed opportunity for children's health-related physical activity in motorized societies. PMID- 12568481 TI - A multimethod approach to the evaluation of community preschool speech and language therapy provision. AB - OBJECTIVES: The paper presents a research study investigating the effectiveness and acceptability of community speech and language therapy provision for preschool children with early speech/language delays. As a 'worked example', it demonstrates the value of a multimethod approach to evaluation. METHODS: The paper examines how the research findings of a pragmatic randomized controlled trial (RCT), a survey questionnaire and qualitative in-depth interviews, which were the methods used in the evaluation, overlap and complement each other. RESULTS: There was little evidence to establish the effectiveness of the speech and language therapy provided in the trial. This lack of difference was reflected in responses of parents to items on the questionnaire. The findings of the RCT, questionnaire and interviews all cast considerable doubt on the prospect of spontaneous resolution of the children's difficulties. Although the RCT showed few differences between the children allocated to immediate therapy or 'watchful waiting', the questionnaire and interviews revealed the circumstances in which the parents felt that these intervention strategies had been acceptable and unacceptable. CONCLUSIONS: Although the trial provided information about the progress of the children, the questionnaire and interview components highlighted the advantages and limitations of the intervention from the viewpoint of parents, thereby helping to explain the RCT findings. Thus, the study demonstrates how a multimethod approach to evaluation can yield useful information to explain the findings of RCTs. PMID- 12568482 TI - The role of disability living allowance in the management of attention deficit/hyperactivity disorder. AB - OBJECTIVE: To explore the use of Disability Living Allowance (DLA) by families of children and adolescents with Attention-Deficit/Hyperactivity Disorder (ADHD), and to discuss the implications for clinicians involved in their treatment. STUDY DESIGN: Opportunistic survey of patients attending ADHD clinic. SETTING: Urban area in the north-east of England. SUBJECTS: A total of 32 carers of children being treated for ADHD with methylphenidate. INTERVENTION: Semi-structured telephone interviews about receipt and use of DLA. This involved open and closed questions and a multiple-choice section. RESULTS: In total, 19 out of the 32 families were receiving DLA. They chose to use it mainly to replace clothes and furniture and to provide diversions and activities for the children concerned. Some families were unaware of potential eligibility for DLA, whereas a few had chosen not to apply. Only one family's application for DLA had been unsuccessful. Carers were unanimously positive about the extra income. CONCLUSIONS: Families view DLA as an important means to replace damaged items and to fund recreational activities to contain over-activity. Families receive little formal guidance on ways of using DLA money to support children with ADHD. Virtually no specific training in benefits awareness is provided to general practitioners and child health specialists, who are often asked to judge the child's level of impairment or incapacity. Applying for DLA may affect the therapeutic relationship for good or ill. There is a need for professionals in contact with children with ADHD to inform families of the possibility of receiving DLA and support them in applications. As diagnosis and treatment of ADHD becomes more commonplace, more families are likely to be entitled to claim DLA. This has definite implications for the social security budget. PMID- 12568483 TI - Gender identity problems in autistic children. AB - Several psychiatric and behavioural problems have been described in children with autism. The aim of this case report is to discuss the developmental pattern of cross-gender behaviour in children with autistic disorder. Two boys are described with high functioning autism, followed for about 4 years, who showed persistent gender identity problems. PMID- 12568484 TI - Mucus hypersecretion in respiratory disease. Chair's introduction. PMID- 12568485 TI - Epidemiological studies in mucus hypersecretion. AB - Respiratory mucus in epidemiology has mainly been studied using standardized questionnaires including questions on cough and phlegm. In chronic obstructive pulmonary disease (COPD) much controversy exists regarding the importance of mucus hypersecretion. From being the key element in the 'British hypothesis' it was reduced to being an innocent disorder in the 1980s but is now again recognized as a potential risk factor for an accelerated loss of lung function. Whereas early studies in mainly occupational cohorts showed no effect of chronic mucus hypersecretion on decline in lung function, such an effect has been shown in subsequent studies on general population samples. Chronic mucus hypersecretion also increases risk of hospital admission which may be due to an increased risk of lower respiratory tract infection. In severe COPD this may explain the increased mortality associated with the presence of mucus. In asthma recent findings suggest that in epidemiology chronic mucus hypersecretion may indicate lack of control which leads to an accelerated loss of lung function and increased mortality in subjects with self-reported asthma. PMID- 12568486 TI - Post-secretory fate of host defence components in mucus. AB - Airway mucus is a complex mixture of secretory products that provide a multifaceted defence against infection. Among many antimicrobial substances, mucus contains a peroxidase identical to milk lactoperoxidase (LPO) that is produced by goblet cells and submucosal glands. Airway secretions contain the substrates for LPO, namely thiocyanate and hydrogen peroxide, at concentrations sufficient for production of the biocidal compound hypothiocyanite, a fact confirmed by us in vitro. In vivo, inhibition of airway LPO in sheep significantly inhibits bacterial clearance, suggesting that the LPO system is a major contributor to host defences. Since secretory products including LPO are believed to be steadily removed by mucociliary clearance, their amount and availability on the surface is thought to be controlled solely by secretion. In contrast to this paradigm, new data suggest that LPO and other substances are retained at the ciliary border of the airway epithelium by binding to surface associated hyaluronan, thereby providing an apical, fully active enzyme pool. Thus, hyaluronan, secreted from submucosal gland cells, plays a previously unrecognized pivotal role in mucosal host defence by retaining LPO and possibly other substances important for first line host defence at the apical surface 'ready for use' and protected from ciliary clearance. PMID- 12568487 TI - Mechanisms of submucosal gland morphogenesis in the airway. AB - Submucosal glands (SMGs) are thought to play an important role in the pathogenesis of a number of hypersecretory lung diseases including cystic fibrosis, asthma, and chronic bronchitis. In such diseases, severe SMG hypertrophy and hyperplasia is characteristic of disease progression. Our laboratory has focused efforts on defining both the mechanism of SMG morphogenesis and the identification of SMG stem cells. To this end, we have identified a transcription factor (LEF1) that is temporally and spatially uniquely regulated in SMG progenitors during the initial stages of gland development. LEF1 expression is absolutely required for SMG development in mouse and ferret tracheas, but is insufficient to induce de novo gland development in the absence of other unknown co-factors. In an effort to delineate the transcriptional cascades responsible for inducing LEF1 expression and subsequent SMG development in the airway, we have begun to dissect the regulation of the LEF1 promoter using cell line and transgenic mouse models. Current efforts are focused on defining the cis-acting elements and transcriptional binding factors responsible for Wnt induction of the LEF1 promoter and determining whether the Wnt/beta catenin cascade plays a role in submucosal gland development in vivo. PMID- 12568488 TI - Mucin-producing elements and inflammatory cells. AB - Airway goblet cells and submucosal glands form the major sources of human respiratory mucins. In the adult, mucus-secreting glands occupy about one-third of the inner airway wall wherever there is supportive cartilage (i.e. from the larynx to small bronchi). In hypersecretory conditions such as chronic bronchitis, asthma and cystic fibrosis, glands are considered to be the major source of tracheobronchial mucus, especially that which is expectorated abnormally as sputum. In contrast, goblet cells are regularly found throughout the tracheobronchial tree. Normally sparse or absent in bronchioles (i.e. small airways of less than 1 mm diameter), goblet cells appear and increase in number in airway hypersecretory conditions: their secretions likely contribute to airflow obstruction and early closure of bronchioles, especially during expiration. The increase in gland mass has been considered to be the histological correlate of mucus-hypersecretion in conditions such as chronic bronchitis. However, there appears to be a better association of sputum production with scores of airway wall inflammation than with gland size per se. Thus, while the absolute mass of mucus-secreting tissue is important, it is likely that the release of inflammatory cell secretions (e.g. neutrophil elastase, mast cell chymotryptase), mediators of inflammation (e.g. interleukin 4, 13) and products of the metabolism of arachidonic acid (such as 15-HETE) contribute more than previously realized to the hypersecretion of mucus in chronic bronchitis. New data discussed herein provide supportive evidence for this hypothesis and identify a newly reported link between plasma cells and mucus-hypersecretion by submucosal glands. These considerations demonstrate the complexity of targets that need to be considered for the treatment of mucus hypersecretion. PMID- 12568489 TI - Respiratory tract mucins: structure and expression patterns. AB - Goblet cells produce mainly MUC5AC, but also MUC5B and some MUC2 in apparently 'irritated' airways. MUC5B dominates in the submucosal glands although a little MUC5AC and MUC7 are usually present. MUC4 originates from the ciliated cells. After separation into a gel and a sol phase, lysozyme and lactoferrin are enriched in the salivary gel phase suggesting that mucus may act as a matrix for 'protective' proteins on the mucosal surface. A salivary MUC5B N-terminal fragment consistent with a cleavage event in the D' domain was detected with antibodies against various N-terminal peptide sequences suggesting that assembly of MUC5B occurs through a mechanism similar to that of the von Willebrand factor. Identification of additional cleavage sites C-terminal to the D' domain suggests that most of the N-terminal low-glycosylated part of MUC5B may be removed without affecting the oligomeric nature of the mucin. Possibly, the generation of mucins with different macromolecular properties through proteolytic 'processing' is one way of adapting the mucus polymer matrix to meet local physiological demands. Monomeric mucins that appear to turn over rapidly in the airway epithelium have been identified using radiolabelled mucin precursors. 'Shedding' of such mucins after microbe attachment may prevent colonization of epithelial surfaces. PMID- 12568490 TI - Development and validation of a lectin-based assay for the quantitation of rat respiratory mucin. AB - The significance of a mucus hypersecretory phenotype to the pathogenesis of airways disease is controversial, but increasing evidence suggests that it can negatively impact upon patient health status. A critical aspect of developing our understanding of the role of mucus hypersecretion in disease is the development and appropriate use of methods permitting detection and quantitation of mucins in biological samples. However, the unique biophysical and biochemical properties of this class of glycoproteins do not lend themselves to routine quantitation. Individual pure mucins are not commercially available, the reactivity profile of commonly used reagents is generally not well characterized and assay development and validation is rarely covered adequately in the literature. Therefore quantitation of mucin in biological samples relies upon careful histochemical and biochemical characterization and partially purified mucin preparations. The absence of tools considered essential for assay development in other areas means that this characterization process does not generally lead to proof of selective detection of mucin, but rather to a level of confidence that mucin is detected and defined contaminants are not. This chapter provides an example of the process of development and validation of a lectin-based assay for quantitation of mucin in untreated complex biological samples. PMID- 12568491 TI - Regulation of mucin secretion from in vitro cellular models. AB - Conceptually, in vitro models for airway mucin secretion may provide useful information pertinent to many aspects of goblet cell biology/physiology. Such models may be especially useful in identifying potential secretagogues, probing the distribution of receptors between goblet cell apical and basolateral membrane domains, and revealing intracellular messenger pathways underlying receptor activation. We have focused most recently on human bronchial epithelial cell cultures grown as tracheal xenografts and SPOC1 cell cultures. These two models are remarkably similar with respect to the regulation of mucin secretion: luminal challenges with the P2Y2 purinoceptor agonists ATP or UTP elicit mucin secretion with EC50s of about 3 microM and archetypal agonists to other purinoceptors test negative. P2Y2 purinoceptors typically couple via Gq to phospholipase C, suggesting that intracellular Ca2+ and protein kinase C (PKC) are important in activating intracellular pathways leading to goblet cell mucin release. Consistent with this notion, phorbol myristate acetate and ionomycin elicit mucin secretion from SPOC1 cells and HBE xenografts, whereas cyclic nucleotides do not. Delineation of the molecules comprising these receptor/messenger interactions and their supporting pathways remains an important challenge for the development of drugs effective in therapeutic interventions in mucin hypersecretory airway diseases; with these models we have initiated the process. PMID- 12568492 TI - Oscillations of pH inside the secretory granule control the gain of Ca2+ release for signal transduction in goblet cell exocytosis. AB - Although Ca2+ plays a critical function in relaying intracellular messages, the role of subcellular organelles in the dynamics of intracellular Ca2+ still remains largely unexplored. We recently demonstrated that secretory granules can signal their own export from the cell by releasing Ca2+ to the cytosol. Oscillations and release of Ca2+ in/from the granule result from the combined action of a Ca2+/K+ ion exchange process that occurs in the granule's matrix, and the sequential activation of two Ca2+-sensitive ion channels: an inositol 1,4,5 trisphosphate receptor Ca2+ channel (InsP3R) and an apamin-sensitive Ca2+ activated K+ channel (ASK(Ca)). The results reported here from studies using isolated mucin granules indicate that intralumenal granular Ca2+ oscillations ([Ca2+]L) and the corresponding cyclical release of Ca2+ to the cytosol induced by InsP3 are accompanied by corresponding intragranular pH(G) oscillations. Our data show that K+-induced unbinding of Ca2+ from the mucin matrix increases as the pH(G) declines. These observations suggest that oscillations of pH(G) can modulate the gain of the Ca2+/K+ ion exchange process, thereby controlling the amplitude of [Ca2+]L oscillations and the granule-cytosol release gradient of [Ca2+]. PMID- 12568493 TI - A calcium-activated chloride channel blocker inhibits goblet cell metaplasia and mucus overproduction. AB - We have previously shown that expression of a Ca2+-activated Cl- channel (mCLCA3 in mice and bCLCA1 in humans) is up-regulated along with goblet cell metaplasia and mucus overproduction in the lungs of interleukin 9 (IL9) transgenic mice, and in human primary lung cultures by IL4, IL13 and IL9. We show here that hCLCA1 expression in NCI-H292 cells specifically induces soluble gel-forming mucin production. Moreover, niflumic acid (NFA), a blocker of hCLCA1-dependent Cl- efflux, inhibits MUC5A/C production in these cells. NFA treatment during natural antigen-exposure, where mCLCA3 is greatly up-regulated in the lung, significantly reduces airway inflammation, goblet cell metaplasia and mucus overproduction in vivo. These data suggest that this Ca2+-activated Cl- channel plays an important role in epithelial-regulated inflammatory responses, including goblet cell metaplasia, and represents a potential novel therapeutic target for the control of mucus overproduction in chronic pulmonary disorders. PMID- 12568494 TI - Mechanisms by which gram-positive bacteria and tobacco smoke stimulate mucin induction through the epidermal growth factor receptor (EGFR). AB - Mucin, the major macromolecular component of mucus, is generally considered to be a protective substance. When overproduced in a variety of lung diseases, however, mucin gives rise to clinical problems such as airway obstruction and recurrent infection. Our approach to identifying drug targets for the control of mucin overproduction is the analysis of cellular signalling pathways linking stimuli in the diseased lung to mucin transcription. Here we show that mucin transcription in response to both gram-positive bacteria and tobacco smoke is mediated through activation of the epidermal growth factor receptor (EGFR). The mode of activation of EGFR in response to bacterial lipoteichoic acid involves cleavage of the transmembrane ligand HBEGF by ADAM 10, whereas the activation of EGFR in response to smoke involves cleavage of amphiregulin by ADAM 17. PMID- 12568495 TI - Non-allergic models of mucous cell metaplasia and mucus hypersecretion in rat nasal and pulmonary airways. AB - Mucous cell proliferation and hypersecretion of airway mucus are important pathological features of human respiratory disorders such as asthma and chronic bronchitis. In addition to airborne allergens and infectious agents, inhaled chemical irritants such as ozone and cigarette smoke have been demonstrated to induce changes in airway mucus production. Cellular and molecular mechanisms involved in non-allergic, toxicant-induced mucous cell metaplasia (MCM; transformation of airway epithelium, normally devoid of mucous cells, to secretory epithelium containing numerous mucus-secreting cells) are still unclear. We have used two experimental models of toxicant-induced MCM in the airways of rats to study the epithelial and inflammatory factors involved in the pathogenesis of MCM. Mucin-specific gene expression and MCM are induced in the nasal transitional epithelium (NTE), but not in the bronchiolar epithelium of F344 rats acutely exposed to ozone, an important air pollutant of photochemical smog. Inhalation of endotoxin, a lipopolysaccharide-protein molecule of gram negative bacteria, induces MCM in the bronchiolar epithelium, but not in the NTE, of rats. Both ozone- and endotoxin-induced MCM are dependent on neutrophilic inflammation. Interestingly, each toxicant enhances the MCM induced by the other toxicant. These synergistic effects elicited by coexposure to ozone and endotoxin are also mediated, in part, by neutrophils. PMID- 12568496 TI - Cytokine regulation of mucus production in a model of allergic asthma. AB - Mucus hyperproduction in asthma results from airway inflammation and contributes to clinical symptoms, airway obstruction and mortality. Th2 lymphocytes and eosinophils dominate the airway inflammatory infiltrate. We investigated the role of different lymphocyte subsets and their cytokines in the stimulation of mucus production using a system in which T cell receptor (TCR) transgenic CD4+ Th cells were generated in vitro, transferred into recipient mice and activated in the respiratory tract with inhaled antigen. Th2 cells induced mucus production and eosinophilic inflammation, while mice that received Th1 cells exhibited airway inflammation without mucus. Th1 cells failed to stimulate mucus due to the inhibitory effects of interferon (IFN)gamma. Mucus was induced by Th2 cells in the absence of interleukin (IL)4, IL5, eosinophils and mast cells, but not without IL4R alpha signalling. Th2 cells lacking IL13 could not stimulate mucus production, despite the presence of airway inflammation. IL9 also stimulates mucus through an IL13-mediated pathway. Using bone marrow chimeras we show that IL13 acts on structural cells in the lung, most likely by direct stimulation of epithelial cells, and not through intermediate inflammatory cells. In asthma, airway inflammation with CD4+ Th2 cells stimulates mucus production by a single pathway mediated by IL13. PMID- 12568497 TI - The role of apoptotic regulators in metaplastic mucous cells. AB - Exposure of airways to environmental toxins or allergens induces proliferation of epithelial cells. Depending on the type of exposure, existing and newly formed cells can differentiate into mucus-producing cells resulting in mucous cell metaplasia (MCM). During recovery, the epithelium reduces the number of epithelial cells to return to the original state. Understanding the mechanisms involved in this resolution could be useful in deleting mucous cells and, thereby, mucous secretions. We have found that metaplastic mucous cells induced by exposure to ozone, endotoxin, cigarette smoke or allergens in epithelia of various regions of the airways express Bcl-2, a regulator of apoptosis, and neutrophils appear to be involved in its expression. The percentage of Bcl-2 positive mucous cells is decreased prior to the resolution of MCM. Furthermore, targeted reduction of Bcl-2 expression causes a dose-dependent reduction of epithelial mucous cells, suggesting that Bcl-2 is involved in maintaining metaplastic mucous cells. Horses with recurrent airway obstruction show an increased percentage of Bcl-2-positive mucous cells compared to their normal counterparts. These studies suggest that down-regulation of Bcl-2 expression may be useful to reduce mucous secretions in diseased subjects. The role of Bax in the reduction of MCM during prolonged exposure to allergen is also discussed. PMID- 12568498 TI - Current and future therapies for airway mucus hypersecretion. AB - Mucus hypersecretion is a prominent feature of chronic inflammatory diseases of the airways, including asthma, chronic obstructive pulmonary disease (COPD) and cystic fibrosis, but little is known about the effects of current therapies for airway disease because of the difficulties in quantifying mucus hypersecretion in clinical studies. Anticholinergics may reduce mucus hypersecretion, whereas beta2 agonists and mucolytics have little obvious effect. Corticosteroids are highly effective in inhibiting mucus hypersecretion in asthma by suppressing the underlying inflammatory process, but are ineffective in COPD and cystic fibrosis. Novel approaches in the future may include inhibition of sensory neuropeptides by tachykinin antagonists, modulators of sensory nerves or K+ channel openers. Inhibition of Th2 cytokines (interleukin [IL]4, IL9, IL13) may also be effective in asthma. In COPD inhibition of neutrophil-derived proteases by small molecule inhibitors or inhibiting neutrophilic inflammation in the airways by reducing neutrophil chemotaxis may also be effective strategies. Several novel targets involved in mucus hypersecretion have recently been identified, including epidermal growth factor receptors, MARCKs, Ca2+-activated Cl- channels and mitogen-activated protein kinases. However, the clinical benefits from inhibiting mucus hypersecretion are still not certain, casting some doubts on this therapeutic approach. PMID- 12568499 TI - Clinical evaluation of new therapies for treatment of mucus hypersecretion in respiratory diseases. AB - In the past mucoactive drugs in airway diseases have been identified and profiled in symptom-based animal experiments and in clinical trials along related lines (cough and expectoration). Presently available drugs of this class are not generally accepted by licensing authorities worldwide and no new molecule clinically profiled as a mucoactive drug has been brought to regulatory approval in the past 20 years. Among regulatory guidelines only the CPMP 1999 'points to consider' on drug development in chronic obstructive pulmonary disease (COPD) advises for mucoactive drug development by suggesting that an indication for symptomatic treatment may be established on the basis of a symptom-related primary endpoint that should be justified as for its importance and supported by a co-primary lung function endpoint. Quality and safety of the new drug must be documented in long-term studies and the indication and use clearly described based on established or adequately profiled new primary endpoints in two pivotal studies. Published trials on mucoactive drugs have used a variety of endpoints. These include mucus hypersecretion-related symptoms by questionnaire, expectorated volume and dry weight, and mucus viscosity, elasticity and transportability. Most methods and endpoints are not validated and a positive standard of treatment is not established. New surrogate markers of efficacy for shorter term trials, e.g. induced or spontaneous sputum based assays (cellularity, mucus antigens), exhaled breath (NO), breath condensate (eicosanoids) or airway biopsy are only partially validated and the risk of false positive or negative phase II results is appreciably high. On the other hand, lung function measurements including airway hyper-reactivity assessment and typical phase III (long-term) endpoints like dyspnoea ratings, health status assessments, incidence of exacerbations and lung function decline over time are validated endpoints and offer a high likelihood of regulatory acceptance. Proof for no depression of lung mucociliary clearance is an important safety endpoint. PMID- 12568500 TI - Maltreatment issues and outcomes of adolescents enrolled in substance abuse treatment. AB - Although they are mandated reporters and work regularly with youth who have experienced abuse and trauma, many substance abuse treatment programs do not systematically screen for or address child maltreatment issues. This special issue provides a collection of articles that consistently demonstrate the feasibility of systematically screening for victimization among adolescents entering these programs and documents that multiple co-occurring types of victimization are actually the norm. Other articles in the section examine how levels of abuse and traumatic victimization are associated with aspects of adolescent substance use disorders, traumatic distress symptoms, co-occurring somatic and health problems, HIV risk behaviors, and a victim-to-abuser spiral. Finally, the articles address how this might affect treatment matching across levels of care. This introduction provides an overview of the articles and then highlights some of the key implications of this collective body of work for practice, policy, and future research. PMID- 12568501 TI - Treatment processes and outcomes among adolescents with a history of abuse who are in drug treatment. AB - This study reports on the effects of having a history of physical and/or sexual abuse on treatment processes and posttreatment abstinence among adolescents (N = 803) in a multisite study of drug treatment outcomes. A history of physical and/or sexual abuse was reported by 59% of the girls and 39% of the boys and was related to being White and having higher levels of comorbidity, exposure to other traumatic events, dependence on alcohol and marijuana, problem recognition, and negative peer or family influence. Abused adolescents had more service needs at treatment admission and there was some variation in treatment processes related to gender and abuse status. Youths with a history of physical abuse had a lower likelihood of posttreatment abstinence, except among those who had better rapport with their counselors. Attention to treatment processes and engagement strategies is crucial for treating youths in drug treatment who have a history of abuse. PMID- 12568502 TI - Gender differences in victimization severity and outcomes among adolescents treated for substance abuse. AB - This article uses data from the Global Appraisal of Individual Needs (GAIN) on 214 adolescents entering substance abuse treatment. The goals of the article are to validate the General Victimization Index (GVI), examine its relationship to gender and co-occurring problems, and determine its relationship to outcomes. The GVI includes 15 items on lifetime traumatic events, traumagenic factors, and current worries of victimization. The items fall along a severity dimension (alpha = .88), and evidence was generated to support the construct validity of cutoff scores for interpretation. Girls were significantly more likely than boys to have experienced a variety of victimization incidents. When used as grouping variables, gender and severity of victimization significantly interacted with measures of intake status and were significant predictors of 3-month postdischarge treatment outcomes. PMID- 12568503 TI - Maltreatment issues by level of adolescent substance abuse treatment: the extent of the problem at intake and relationship to early outcomes. AB - Differences in self-reported victimization and outcomes for residential (n = 114) vs. outpatient (n = 73) substance abuse treatment samples were examined. Repeated measures MANOVAs for victimization level by level of care were performed on days of alcohol and marijuana use and substance-related problems. Residential treatment participants reported higher prevalence of victimization and higher levels of general victimization but did not differ on acute (high) victimization at intake. Analyses revealed a significant interaction between follow-up outcomes and level of care for adolescents with acute intake victimization. Adolescents placed in residential treatment were more likely to reduce their days of alcohol and marijuana use and past month substance-related problems at follow-up. Adolescents with low intake levels of victimization did not differ by level of care. Findings suggest that clinicians must carefully weigh placement recommendations for adolescents with maltreatment histories and that researchers should study ways to increase outpatient treatment effectiveness for these adolescents. PMID- 12568504 TI - Traumatic stress and gender differences in relationship to substance abuse, mental health, physical health, and HIV risk behavior in a sample of adolescents enrolled in drug treatment. AB - Research on traumatic stress (TS) among adolescent substance users is limited, with research indicating that not all adolescents who experience trauma are substance users and not all adolescent substance users report symptoms of TS. In the general adolescent population, research on TS symptoms indicates gender differences, with more females reporting traumatic life events and more symptoms associated with traumatic stress. A gap in research exists, however, with regard to gender differences among adolescent substance users who report low versus acute levels of TS symptoms. This study included 274 male and 104 female adolescents enrolled in four drug treatment programs in Arizona. Comparisons between males and females and those with low versus acute levels of TS symptoms were examined with regard to substance use, mental health, physical health, and HIV risk-taking behavior. Results indicate significant differences between males and females and between those reporting low versus acute TS. In general, females and those with acute levels of TS symptoms had higher levels of substance use, mental health, and physical health problems as well as greater HIV risk behaviors when compared to males and those with low levels of TS symptoms. Results of this study indicate the need to assess adolescents for TS, including victimization and maltreatment histories, when entering substance abuse treatment and the need to simultaneously address issues of substance use, TS, and related mental health, physical health, and HIV sex risk behavior while in treatment. PMID- 12568505 TI - Posttreatment victimization and violence among adolescents following residential drug treatment. AB - This article examines the relationships among experiences of childhood abuse, psychiatric disorders, self-reported victimization, and violent behavior, with a focus on gender differences. Data were obtained from treatment entry and 5-year post-treatment interviews of 446 adolescent clients in therapeutic community (TC) drug treatment programs throughout the United States and Canada. Fifty-eight percent of the sample indicated that they engaged in serious violent behaviors (e.g., beatings, threatening or using weapons against other people, or violent crimes such as assaults, rapes, murders) in the 5 years following their separation from TC treatment. Multivariate logistic regression analyses revealed that victimization in the posttreatment period was the most significant factor associated with violent behavior, and pretreatment childhood abuse experiences and psychiatric disorders were not significantly related to the odds of violent behavior. There were significant gender differences in self-reported victimization and violent behavior. The findings suggest that violence in young adulthood for males is related to increasing involvement in violent lifestyles that include drug trafficking, while violence among females is associated with the social and psychological consequences of drug involvement and victimization. High rates of violent involvement and victimization among former adolescent clients suggests the utility of incorporating interventions such as safety oriented strategies for females or interventions that address involvement in the drug use lifestyles (i.e., use and dealing) for both males and females into residential treatment to reduce the likelihood of future violence. PMID- 12568506 TI - Re: Carnes et al. (2001), Extended forensic evaluation when sexual abuse is suspected: a multisite field study, Child Maltreatment, 6(3), 229-241. PMID- 12568507 TI - eHealth--the road ahead. PMID- 12568508 TI - Scrotal swellings. PMID- 12568509 TI - Where there is no ultrasound: delay and failure of antenatal diagnosis of twin pregnancy. AB - We investigate the problem of late diagnosis of twin pregnancy in Soweto, South Africa, where routine antenatal ultrasound is not available. One hundred consecutive pairs of twins were studied, using the notes of mothers who delivered twins at Chris Hani Baragwanath Hospital and the referring Soweto clinics. A positive history was found in 31 mothers (22 family history, nine previous history of twins). Six mothers did not attend for antenatal care. Twenty-five twin pregnancies were discovered at delivery, 15 of them in the second stage of labour, and 27 were diagnosed accidentally in the third trimester. Only 15 pregnancies were referred specifically for suspicion of twin pregnancy. Most twin pregnancies are detected only in the third trimester or at delivery. Until routine ultrasound is available to all pregnant women, the teaching of antenatal care in South Africa must give emphasis to clinical suspicion of twin pregnancy. PMID- 12568510 TI - The diagnosis of extrapulmonary tuberculosis in Malawi. AB - There is little information on a country-wide basis in sub-Saharan Africa about how the diagnosis of extra-pulmonary tuberculosis (EPTB) is made. A country-wide cross-sectional study was carried out in 40 non-private hospitals in Malawi which register and treat (TB) patients in order to assess diagnostic practices in adults registered with EPTB. All patients aged 15 years and above in hospital on treatment for EPTB were reviewed usingTB registers, case note files and clinical assessment. There were 244 patients, 132 men and 112 women whose mean age was 36 years. In 138 (57%) patients, all appropriate procedures and investigations, commensurate with hospital resources, had been carried out. Of 171 EPTB patients with cough for 3 weeks or longer, 138 (81%) submitted sputum specimens for smear microscopy of acid-fast bacilli (AFB). A confirmed diagnosis ofTB was made in 15 (6%) patients based on finding AFB or caseating granulomas in specimens. In 157 (64%) patients, the diagnosis of EPTB was considered to be correct. In 46 (19%) patients the diagnosis was considered to be TB, although different from the type of EPTB with which the patient was registered. In 39 (16%) patients an alternative non-TB diagnosis was made and in two (1%) patients it was not possible to make a decision. Diagnostic practices need to be improved, and ways of doing this are discussed. PMID- 12568511 TI - 324 cases of misplaced IUCD--a 5-year study. AB - Copper-T remains the mainstay of family planning measures in developing countries but is associated with serious complications like bleeding, perforation and migration to adjacent organs or omentum. Patients with misplaced intrauterine contraceptive device (IUCD) may present with pregnancy or lost strings or may remain asymptomatic for years. In our series of 324 cases with misplaced IUCD1; in 258 (79.93%) cases copper-T was found in the uterine cavity and in 47 cases (14.51%) it was removed from cervical canal. In only 18 cases (5.56%), it was translocated and of these 66.67% were inserted at primary health centres. Laparoscopic retrieval was successful in 61.11% cases. Endoscopy thus helps in the localization and retrieval of misplaced IUCDs. A regular follow up of IUCDs for visible threads would help in earlier detection of misplaced IUCDs. Proper training of paramedical staff is mandatory in developing countries to provide safe and better family planning services. PMID- 12568512 TI - Contribution to the surgical treatment of patients with cervical spine cord injury using simple procedures. AB - Thirty-nine patients with acute cervical spine fractures and/or dislocations between C3 and C7 were submitted to an anterior approach using bone graft fixation without screw and plate systems and three required a preliminary posterior approach to reduce a dislocation. Graft dislodgement due to technical problems occurred at a rate of 7.7% postoperatively and 2.8% 1 month later. No redislodgement occurred. All fusions became solid after 3 months. Their progress was based on the Frankel scale, before surgery, at the moment of the discharge, and at 6 months follow-up. This experience shows how patients with an acute cervical injury can improve even when admitted late after trauma. PMID- 12568513 TI - A practical scheme for the initial out-patient management of type-2 diabetes mellitus. PMID- 12568514 TI - Admission troponin T as a prognostic marker and it relationship to streptokinase treatment patients with acute myocardial infarction. AB - The relationship between the admission troponinT (TnT) level and the response to streptokinase (SK) was examined in 76 patients with acute myocardial infarction (AMI). Of 27 TnT positive patients, 10 (37%) showed a response to SK as suggested by a non-invasive criterion for reperfusion, while 24 (49%) were 'responders' among 49 TnT negative patients. There appeared to be a trend towards a better response to SK in the TnT negative group but the difference lacked statistical power due to the small sample size. The mean time-interval between the onset of symptoms and thrombolytic treatment among TnT positive 'non-responders' was significantly (P < 0.005) higher than the TnT negative 'non-responders' (5.23 + 3.42 h versus 2.38 +/- 1.37 h). An 18 month follow up on 61 patients revealed a higher mortality (33%) among TnT positive patients than TnT negative patients (10%). Mortality among TnT positive 'non-responders' was significantly higher (P = 0.0494) than mortality among TnT-negative 'non-responders' (43% versus 9%), indicating that TnT positive patients, non-responsive to SK were at a greater risk of cardiac death. The data suggest that the admission TnT level can be of value in risk stratification of patients with AMI. PMID- 12568515 TI - Alternative medicine use at Vila Central Hospital Vanuatu: a survey of the use of 'custom medicine' in patients and staff. AB - A structured questionnaire was developed to assess the use of custom (traditional or herbal) medicine in 50 medical and 50 surgical inpatients and 50 staff at Vila Central Hospital, Vanuatu. Eighty-six per cent of those interviewed had used custom medicine at least once, 60% within the last year. Twenty-one medical patients (42%) had used custom medicine for their current problem, 12% whilst in hospital. Eighteen surgical patients (36%) had used custom medicine for their current problem, 6% whilst in hospital. Fourteen of the staff (28%) interviewed had used custom medicine within the last year. Sixty-nine per cent of interviewees stated that custom medicine had improved their condition. Whilst custom medicine use was common across all attained education levels, there was a trend for those with higher levels of education to use less custom medicine. The most common form of custom medicine was leaf tea, followed by leaf applications, cutting, magic, massage and chanting. The majority of these were made/prescribed by local 'Klevas' (custom doctors). The cost of the 'medicine' was often substantial, generally many times the cost of a day in hospital. These findings indicate a high prevalence of custom medicine use within Vanuatu. Clinicians must be alert to the possibility of their patients using custom medicine even whilst in hospital and the potential for custom medicine to contribute to the clinical problems. Late presentation of patients to hospital is common in Vanuatu, and the use of custom medicine is partially a contributing factor to this. It also poses questions as to patient confidence in health services. PMID- 12568516 TI - Cerebrovascular diseases in Nigeria: what do we know and what do we need to know? PMID- 12568517 TI - Excision biopsy of lymph node--needle fixing technique. PMID- 12568518 TI - Reagent strip testing of cerebrospinal fluid. PMID- 12568519 TI - A guide to sending e-mail telemedicine referrals. AB - This article describes simple e-mail protocols that have proven their effectiveness in the telemedicine links established by the Swinfen Charitable Trust. They are presented here as a guide for the benefit of those doctors who may be considering setting up their own telemedicine links. PMID- 12568520 TI - Telemedicine--the way ahead for medicine in the developing world. AB - An e-mail link with the facility to send high-resolution digital images is a cheap and uncomplicated telemedicine method. The Swinfen Charitable Trust helped establish such a link in Patan Hospital Kathmandu, Nepal in March 2000. Over 12 months using this link 42 telemedicine referrals were sent to specialists throughout the world. Referrals were: 36% respiratory medicine; 21% neurology, 21% dermatology; 14% cardiology; 5% nephrology; and 3% radiology--28 had digital pictures attached, of which 96% were of high enough quality on which specialists were able to comment. Thirty-nine replies were received. The average time for a specialist reply was 2 days, and 45% were answered within 24 hours. All replies were judged by independent assessors to be helpful or very helpful for diagnosis, management and education. The assessors decided that in 50% of cases the advice if acted upon would have shortened hospital stay. This pilot study has shown that a low-cost telemedicine link is technically feasible and can be of significant benefit for diagnosis, management and education in a developing world setting. PMID- 12568521 TI - Correlation between serum-ascites albumin concentration gradient with gastrointestinal bleeding in patients of portal hypertension. AB - Non-availability of endoscopic screening to detect portal hypertensive patients at a high risk of index gastrointestinal (GI) haemorrhage led us to examine the relationship between serum-ascitic albumin concentration gradient (SAAG) measurements and the occurrence of GI haemorrhage in patients with portal hypertension and ascites. Fifty-six consecutive patients of portal hypertension with ascites attending the GI surgery outpatients clinic were divided into two groups on the basis of history: (a) those who had no history of GI bleeding; and (b) those who had an episode of GI bleeding within the past 21 days. Child-Pugh score was calculated for all patients. All the patients were assessed by ultrasonography and SAAG was estimated. Groups (a) and (b) were compared and sensitivity, specificity, positive and negative predictive value of using SAAG as a possible screening test for GI bleeding were calculated. SAAG values correlated significantly with bleeding and splenomegaly. For prediction of bleeding, SAAG had a sensitivity of 100% and specificity of 33.33%, positive predictive value of SAAG for GI bleeding was 71.4% and the negative predictive value was 100%. Estimation of SAAG is possible even in a small, modestly equipped laboratory, and could provide a new means for the identification of high-risk patients for GI bleeding and define patients more accurately for future clinical studies. The results of the present study are encouraging for clinicians working in developing countries and may help in optimizing prophylactic therapy and where available, improving the cost effectiveness of screening endoscopy. PMID- 12568522 TI - Healthcare workers' knowledge of lymphatic filariasis and its control in an endemic area of Eastern India: implications on control programme. PMID- 12568523 TI - The Bangladesh arsenic catastrophe: clinical manifestations. PMID- 12568524 TI - The prevalence and pattern of skin diseases in relation to CD4 counts among HIV infected police officers in Dar es Salaam. AB - Among HIV-infected individuals, skin diseases cause significant morbidity and are frequently the initial indication of immunosuppression. From an on-going cohort study to determine prevalence and incidence of HIV infection among police officers (POs) and their suitability for HIV vaccine trials, a sub-study was carried out to determine the prevalence and pattern of skin diseases among HIV infected POs and relate this to their immunodeficiency status. Consenting HIV infected POs and their age and sex-matched HIV-negative officers were assessed for presence and type of skin diseases at their workplaces. A questionnaire was used for data collection. Immunodeficiency was measured by plasma CD4+ lymphocytes using flow cytometry. Between November 1998 and 31 December 2000, 716 POs were assessed. Overall HIV-1 prevalence was 17.7% (127/716). One hundred and ninety-one POs (26.7%) had at least one skin diagnosis. HIV-infected POs had significantly higher (41.7%) prevalence of skin diseases than HIV-uninfected POs (26.4%), P = 0.002. Fungal infections were common in both HIV-infected and uninfected POs. Among the HIV infected, other common diseases were: herpes zoster (11.8%); pruritic papular eruption (PPE) (7.1%); seborrheic dermatitis (5.5%); and Kaposi's sarcoma (KS) (1.6%). KS and PPE were associated with severe immunodeficiency, with mean absolute (percentage) CD4+ counts of 75.5 cells/microL (4.0%) and 71.7 cells/microL (4.8%), respectively. The values for herpes zoster and seborrheic dermatitis were 271.1 cells/micronL (12.4%) and 206.3 cells/microL (11.3%), respectively. Skin diseases were common among HIV infected POs. PPE and KS are markers of severe immunodeficiency due to HIV. PPE, herpes zoster and KS strongly suggest underlying HIV-related immunodeficiency and patients with these conditions should be counselled and tested for HIV. PMID- 12568525 TI - Injection safety. PMID- 12568526 TI - Calcinosis cutis: a report of four cases. PMID- 12568527 TI - Postpartum foot drop: a report of four cases. PMID- 12568528 TI - Convexity meningioma presenting as postpartum eclampsia. AB - A 37-year-old woman, Para 5(+0) presented with a 1 year history of recurrent convulsions and progressive weakness of the right side of the body. She had been treated for postpartum eclampsia in her last delivery but symptoms recurred 3 months later. Evaluation including computerized tomography scan of the brain suggested a parieto-temporal meningioma, which was completely excised at craniotomy. Histology confirmed this to be a meningioma. The patient was well at 8 months of follow up. The growth of meningiomas may increase during pregnancy due to presence of receptors for progestational hormones in the tumour and the meningioma may become symptomatic in pregnancy, presenting as eclampsia. Close follow up of patients with eclampsia is necessary to identify neurological features that may lead to a diagnosis of meningioma. Early diagnosis is essential if a good outcome is to be ensured. PMID- 12568529 TI - Traumatic diaphragmatic rupture. PMID- 12568530 TI - Upper gastrointestinal bleeding in tropical pancreatitis due to pseudoaneurysm rupture. PMID- 12568531 TI - Acute flaccid paralysis and obligations of clinicians. PMID- 12568532 TI - Treatment of echinococcosis at rural hospital level in Africa. PMID- 12568533 TI - Epidural needle and intraosseous access. PMID- 12568534 TI - Unusual method for tracheal tube placement. PMID- 12568535 TI - Development of a radioimmunoassay procedure for aflatoxin B1 measurement. AB - A radioimmunoassaay (RIA) procedure to measure aflatoxin B(1) (AfB(1)) in agricultural commodities was developed. AfB(1) oxime derivative was synthesized, characterized, and used for preparation of (125)I-labeled AfB(1). Antiaflatoxin B(1) serum was raised in-house using AfB(1)-bovine serum albumin conjugate as immunogen. The assay system was optimized in the range of 0.2-5 ng/mL, using a liquid phase (PEG) as well as a solid phase (coated polystyrene beads) separation system. Inter-assay and intra-assay variations, recovery, and parallelism studies validated the assay. AfB(1) analysis was carried out in nearly 130 samples of different agricultural commodities. The correlation coefficient was determined using commercial ELISA and in-house-developed RIA methods. PMID- 12568536 TI - Determination of the activity of acidic phytate-degrading enzymes in cereal seeds. AB - Five different methods were compared to elucidate the total activity of the acidic phytate-degrading enzymes present in the seeds of rye, wheat, and barley. Phytate-degrading activity was studied at pH 5.0 by quantifying the liberated phosphate. Rye showed the highest acid phytate-degrading activity among the cereals studied. Using an aqueous extract, only 30-50% of the activity was found (rye, 3443 mU g(-1) of grain; wheat, 1026 mU g(-1) of grain; barley, 1032 mU g( 1) of grain) compared to that found by the direct incubation of the dry-milled cereal grains in a buffered phytate-containing solution (rye, 6752 mU g(-1) of grain; wheat, 2931 mU g(-1) of grain; barley, 2093 mU g(-1) of grain). Extending the extraction time resulted in an increase in extractable phytate-degrading activity by, at maximum, 10-15%. Extraction of phytate-degrading activity is strongly enhanced in the presence of Triton X-100 and the protease inhibitor phenylmethylsulfonyl fluoride (rye, 6536 mU g(-1) of grain; wheat, 2873 mU g(-1) of grain; barley, 2023 mU g(-1) of grain), suggesting at least a partial association with membrane structures and a degradation by proteolytic activity during extraction. In addition, it was shown that determining phytate-degrading activity by quantification of the liberated inorganic phosphate is more robust and precise than determining phytate-degrading activity by quantification of the residual phytate. PMID- 12568537 TI - Biomimesis of linolenic acid transport through model lipidic membranes by differential scanning calorimetry. AB - Multienoic fatty acids, such as linolenic acid, show their ability to interact with and to penetrate into model biomembranes by biomimetic experiments performed to support the absorption route followed by n-3 fatty acid in cells. The thermotropic behavior of model biomembranes, that is, dimyristoylphosphatidylcholine multilamellar or unilamellar vesicles, interacting with linolenic acid was investigated by differential scanning calorimetry. When dispersed in liposomes during their preparation, the examined biomolecule was found to interact with the phospholipid bilayers by modifying the gel to liquid crystal phase transition of lipid vesicles; this modification is a function of the fatty acid concentration. Calorimetric analysis was also performed on samples obtained by leaving the pure n-3 acid in contact with lipid aqueous dispersions (multilamellar or unilamellar vesicles) and then examining the thermotropic behavior of these systems for increasing incubation times at temperatures higher than the transitional lipid temperature. Linolenic acid (LNA) was able to migrate through the aqueous medium and successively to interact with the vesicle surface and to penetrate into the model membranes, following a flip-flop mechanism, with a faster and higher effect for unilamellar vesicles, caused by the larger lipid surface exposed, compared to the multilamellar ones, although due to the lipophilic nature of LNA, such a transfer is hindered by the aqueous medium. The relevance of the medium in LNA absorption has been well clarified by other biomimetic transfer experiments, which showed the LNA transfer from loaded multilamellar vesicles to empty vesicles. Taken together, the present findings support the hypothesis of a passive n-3 acid transport as the main route of absorption into cell membranes. PMID- 12568538 TI - Multielement analysis of Canadian wines by inductively coupled plasma mass spectrometry (ICP-MS) and multivariate statistics. AB - Trace element fingerprints were deciphered for wines from Canada's two major wine producing regions, the Okanagan Valley and the Niagara Peninsula, for the purpose of examining differences in wine element composition with region of origin and identifying elements important to determining provenance. Analysis by ICP-MS allowed simultaneous determination of 34 trace elements in wine (Li, Be, Mg, Al, P, Cl, Ca, Ti, V, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Br, Rb, Sr, Mo, Ag, Cd, Sb, I, Cs, Ba, La, Ce, Tl, Pb, Bi, Th, and U) at low levels of detection, and patterns in trace element concentrations were deciphered by multivariate statistical analysis. The two regions were discriminated with 100% accuracy using 10 of these elements. Differences in soil chemistry between the Niagara and Okanagan vineyards were evident, without a good correlation between soil and wine composition. The element Sr was found to be a good indicator of provenance and has been reported in fingerprinting studies of other regions. PMID- 12568539 TI - Simple and rapid determination of thiabendazole, imazalil, and o-phenylphenol in citrus fruit using flow-injection electrospray ionization tandem mass spectrometry. AB - A simple and rapid analytical method for thiabendazole (TBZ), imazalil (IMA), and o-phenylphenol (OPP) in citrus fruit has been developed by using flow-injection electrospray ionization tandem mass spectrometry for the first time. The method involves the combined use of stable isotopically labeled internal standards (thiabendazole-(13)C(6), imazalil-d(5), and p-phenylphenol-d(9)) and a multiple reaction monitoring technique. The average recoveries for the fungicides at the tolerance levels (TBZ and OPP, 10 mg/kg; IMA, 5 mg/kg) ranged from 77 to 101%, with the coefficients of variation (CVs) ranging from 0.7 to 4.2% (n = 5). At half the tolerance levels (TBZ and OPP, 5 mg/kg; IMA, 2.5 mg/kg), the average recoveries ranged from 62 to 112%, with the CVs ranging from 0.7 to 8.4% (n = 5). The CVs of the average recoveries, obtained from lemon samples fortified with three fungicides at the tolerance levels, obtained on three different days over two weeks, ranged within 2%. The analysis time, including sample preparation and determination, is only 15 min. PMID- 12568540 TI - Highly sensitive analysis of iodide anion in seaweed as pentafluorophenoxyethyl derivative by capillary gas chromatography. AB - A simple and sensitive gas chromatography (GC) method is described for the trace analysis of iodide anion (iodide) in processed seaweed as an organic derivative. The method is based on the derivatization of aqueous iodide extracted from seaweed with 2-(pentafluorophenoxy)ethyl 2-(piperidino)ethanesulfonate in toluene using tetra-n-hexylammonium bromide as a phase-transfer catalyst. The resulting pentafluorophenoxyethyl iodide is highly responsive to an electron-capture detector (ECD) and was analyzed by GC-ECD, giving a low detection limit of approximately 2.7 nM (2.7 fmol/microL injected). Interferences of some common anions in the analysis of iodide were studied and proved to be minimal. Application of the method to the analysis of iodide in processed seaweed was performed. PMID- 12568541 TI - Matrix solid-phase dispersion extraction and gas chromatographic determination of chloramphenicol in muscle tissue. AB - A method based on matrix solid-phase dispersion (MSPD) was developed for the gas chromatographic (GC) determination of chloramphenicol (CAP) residues in animal muscle tissue. Muscle tissue was blended with octadecylsilyl-derivatized silica (C(18)). A column made from the C(18)/muscle tissue matrix was washed with n hexane and acetonitrile/water (5 + 95), after which CAP was eluted with acetonitrile/water (50 + 50) and partitioned into ethyl acetate. The final extract was evaporated, and a trimethylsilyl derivative of CAP was prepared with Sylon HTP and detected by GC with an electron capture detector (ECD) and a mass spectrometer. For quantitation, the internal standard used was the meta isomer of CAP (m-CAP) for GC-ECD. Muscle tissue samples were fortified at three concentration levels. At 5, 10, and 15 microg/kg levels the respective mean recoveries were 93, 96, and 98%, and the repeatabilities were 13, 11, and 3%. The detection and quantitation limits with ECD were 1.6 and 4.0 microg/kg, respectively. No statistically significant difference was observed in the efficiency of CAP extraction from muscle tissue of various animals (bovine, porcine, and poultry) by the MSPD technique. PMID- 12568542 TI - Electrochemical and spectroscopic studies of the oxidation mechanism of the herbicide propanil. AB - Electrochemical oxidation of propanil in deuterated solutions was studied by cyclic, differential pulse, and square wave voltammetry using a glassy carbon microelectrode. The oxidation of propanil in deuterated acid solutions occurs at the nitrogen atom of the amide at a potential of +1.15 V vs Ag/AgCl. It was also found that, under the experimental conditions used, protonation at the oxygen atom of propanil occurs, leading to the appearance of another species in solution which oxidizes at +0.60 V. The anodic peak found at +0.79 V vs Ag/AgCl in deuterated basic solutions is related to the presence of an anionic species in which a negative charge is on the nitrogen atom. The electrochemical data were confirmed by the identification of all the species formed in acidic and basic deuterated solutions by means of NMR spectroscopy. The results are supported by electrochemical and spectroscopic studies of acetanilide in deuterated solutions. PMID- 12568543 TI - Supercritical carbon dioxide processing of pyrethrum oleoresin and pale. AB - Possible refining of crude hexane extract (CHE) from pyrethrum flowers and further refining of Pyrethrum Board of Kenya (PBK) pale product is investigated with both liquid and supercritical carbon dioxide. The experiments were carried out in a small pilot plant with a 200 mL extractor and three cyclonic separators in series. To understand the dynamics of pyrethrin extraction, CHE was extracted in a single step; pyrethrin concentration was found to be improved from 0.16 to 0.50 g/g. The effects of temperature and pressure on the quality of the extract were studied at 29 degrees C and 80 bar and at 40 degrees C and 100 bar. Liquid CO(2) processing (29 degrees C, 80 bar) yielded slightly better product quality. A comparison study of CHE and PBK pale processing with supercritical CO(2) (40 degrees C, 100 bar) showed that the final products were similar in terms of pyrethrin content. Extraction of both PBK pale and CHE in two steps with different operating conditions improved their purity. PMID- 12568544 TI - Acaricidal activity of clove bud oil compounds against Dermatophagoides farinae and Dermatophagoides pteronyssinus (Acari: Pyroglyphidae). AB - The acaricidal activity of clove (Eugenia caryophyllata) bud oil-derived eugenol and its congeners (acetyleugenol, isoeugenol, and methyleugenol) against adults of Dermatophagoides farinae and Dermatophagoides pteronyssinus was examined using direct contact application and fumigation methods and compared with those of benzyl benzoate and N,N-diethyl-m-toluamide (DEET). Responses varied according to compound, dose, and mite species. On the basis of LD(50) values, the compound most toxic to D. farinae adults was methyleugenol (0.94 microg/cm(2)) followed by isoeugenol (5.17 microg/cm(2)), eugenol (5.47 microg/cm(2)), benzyl benzoate (9.22 microg/cm(2)), and acetyleugenol (14.16 microg/cm(2)). Very low activity was observed with DEET (37.59 microg/cm(2)). Against D. pteronyssinus adults, methyleugenol (0.67 microg/cm(2)) was much more effective than isoeugenol (1.55 microg/cm(2)), eugenol (3.71 microg/cm(2)), acetyleugenol (5.41 microg/cm(2)), and benzyl benzoate (6.59 microg/cm(2)). DEET (17.85 microg/cm(2)) was least toxic. These results indicate that the lipophilicity of the four phenylpropenes plays a crucial role in dust mite toxicity. The typical poisoning symptom of eugenol and its congeners was a similar death symptom of the forelegs extended forward together, leading to death without knockdown, whereas benzyl benzoate and DEET caused death following uncoordinated behavior. In a fumigation test with both mite species, all four phenylpropenes were much more effective in closed containers than in open ones, indicating that the mode of delivery of these compounds was largely due to action in the vapor phase. Eugenol and its congeners merit further study as potential house dust mite control agents or as lead compounds. PMID- 12568545 TI - Natural fungicides from Ruta graveolens L. leaves, including a new quinolone alkaloid. AB - Bioassay-directed isolation of antifungal compounds from an ethyl acetate extract of Ruta graveolens leaves yielded two furanocoumarins, one quinoline alkaloid, and four quinolone alkaloids, including a novel compound, 1-methyl-2-[6'-(3' ',4' '-methylenedioxyphenyl)hexyl]-4-quinolone. The (1)H and (13)C NMR assignments of the new compound are reported. Antifungal activities of the isolated compounds, together with 7-hydroxycoumarin, 4-hydroxycoumarin, and 7-methoxycoumarin, which are known to occur in Rutaceae species, were evaluated by bioautography and microbioassay. Four of the alkaloids had moderate activity against Colletotrichum species, including a benomyl-resistant C. acutatum. These compounds and the furanocoumarins 5- and 8-methoxypsoralen had moderate activity against Fusarium oxysporum. The novel quinolone alkaloid was highly active against Botrytis cinerea. Phomopsis species were much more sensitive to most of the compounds, with P. viticola being highly sensitive to all of the compounds. PMID- 12568546 TI - Structure-dependent phytotoxicity of catechins and other flavonoids: flavonoid conversions by cell-free protein extracts of Centaurea maculosa (spotted knapweed) roots. AB - Invasive plants are believed to succeed in part by secretion of allelochemicals, thus displacing competing plant species. Centaurea maculosa (spotted knapweed) provides a classic example of this process. We have previously reported that spotted knapweed roots secrete (+/-)-catechin and that (-)-catechin, but not (+) catechin, is phytotoxic and hence may be a major contributor to C. maculosa's invasive behavior in the rhizosphere. In this communication, we explore both structure/activity relationships for flavonoid phytotoxicity and possible biosynthetic pathways for root production of (+/-)-catechin. Kaempferol and dihydroquercetin were shown to be phytotoxic, while quercetin was not. Kaempferol was converted to dihydroquercetin and (+/-)-catechin when treated with total root protein extracts from C. maculosa, but quercetin was not. This finding suggests an alteration in the standard flavonoid biosynthetic pathway in C. maculosa roots, whereby kaempferol is not a dead-end product but serves as a precursor to dihydroquercetin, which in turn leads to (+/-)-catechin production. PMID- 12568547 TI - Antioxidant activity of South African red and white cultivar wines: free radical scavenging. AB - The free radical scavenging activity of South African red (n = 46) and white (n = 40) cultivar wines was determined using 2,2'-azinobis(3 ethylbenzothialozinesulfonic acid) radical cations (ABTS(.+)) and 2,2-diphenyl-1 picrylhydrazyl radicals (DPPH.). The total antioxidant activities (TAA) of red and white wines using ABTS(.+) were 14.916 and 0.939 mM Trolox, respectively, at corresponding total phenol (TP) contents of 2339.0 and 273.8 mg of gallic acid equiv/L. Ruby Cabernet wines had the lowest TAA(ABTS) (13.177 mM Trolox) of the red wines, whereas the TAA(ABTS) values of Chardonnay and Chenin blanc wines were the highest (1.060 mM Trolox) and lowest (0.800 mM Trolox) of the white wines. The TAA(DPPH) values were of the same magnitude as the TAA(ABTS) values, and similar trends were observed. TAA correlated (P < 0.001) with total phenol content of red (r = 0.935) and white (r = 0.907) wines, as well as flavanol content of red wines (r = 0.866) and tartaric acid ester content of white wines (r = 0.767). Canonical discriminant analysis using phenolic composition and antioxidant activity was applied to differentiate between red and white cultivar wines. PMID- 12568548 TI - Variability in Neem (Azadirachta indica) with respect to azadirachtin content. AB - There is a controversy over variations in azadirachtin content in neem (Azadirachta indica) seeds among various provenances and countries. Also, variations in azadirachtins are usually attributed to climatic conditions such as temperature and humidity. The present study was undertaken to evaluate qualitative and quantitative variability in azadirachtins A and B among various neem provenances or individual neem trees. Forty-three provenances of India were examined for intraprovenance variability in azadirachtin A and B content and oil percentage. Twenty-eight individual neem trees from five provenances of different agroclimatic regions were also examined for interprovenance variability. The azadirachtins were quantified using reversed phase analytical HPLC. There were wide variations in oil and azadirachtin contents among different provenances. Azadirachtin A ranged from 556.9 to 3030.8 mg kg(-)(1) of kernels, whereas azadirachtin B was in the range 43.1-590.6 mg kg(-)(1) of kernel among the provenances investigated. Analysis of variance among various neem provenances showed significant differences in oil content, azadirachtin A, total azadirachtin (A + B), and A:B ratio. There were individuals with high and low azadirachtins within a single provenance, and this trend was observed in all of the provenances selected from five agroclimatic regions of the country. Variations among individual trees of a particular provenance indicated that climatic factors such as rainfall, humidity, or temperature did not influence azadirachtin content in the neem trees. The present study shows that there are individual genetic differences among neem trees. A systematic study for tree improvement with a population of mother trees with desired traits should be undertaken by performing half-sib progeny trials and further selections by clonal propagations. The role of genetic makeup needs further research. PMID- 12568549 TI - Effect of organic acids and nitrogen source on alcoholic fermentation: study of their buffering capacity. AB - The effect of tartaric acid and other organic acids on alcoholic fermentation was studied. Organic acids in media with high sugar concentrations and ammonium as the sole nitrogen source had an enormous impact on Saccharomyces cerevisiae metabolism during alcoholic fermentation. The main effect on yeast metabolism was the quick acidification of the media in the absence of organic acids. All of the organic acids used in this study (tartaric, malic, citric, and succinic acids) showed a buffering capacity, but not all of the acids had the same one. However, the results suggested that buffering should not be considered the only effect of organic acids on yeast metabolism. Nitrogen source also had a great influence on media pH. Ammonium consumption by yeasts produced a greater acidification of the media than when amino acids were used. PMID- 12568550 TI - Evolution of sugars in cider brandy aged in oak barrels: a contribution to its characterization. AB - A chemometric study was carried out to typify cider brandies according to the type of wood employed in the maturation process and their aging time. Monosaccharides, previously derivatized with p-aminobenzoic ethyl ester, were analyzed using a reversed-phase high-performance liquid chromatographic method. Univariate data treatment was not sufficient to enable differentiation of the classes of cider brandies on the basis of wood type and maturation time. Two linear combinations of original variables, ascertained by principal components analysis, provided an adequate data structurization. A mathematical decision rule was established to classify cider brandies with prediction capacities of 92 and 97% using an LDA method and Bayesian analysis, respectively. The use of the PLS algorithm allowed the authors to differentiate cider brandies according to the age and type of oak used in the aging process. PMID- 12568551 TI - Effects of gamma-irradiation on the free radical and antioxidant contents in nine aromatic herbs and spices. AB - Nine spice and aromatic herb samples (i.e., basil, bird pepper, black pepper, cinnamon, nutmeg, oregano, parsley, rosemary, and sage) were gamma-irradiated at a dose of 10 kGy according to commercial practices. The effects of the disinfection treatment on the content of organic radicals and some nutrients (namely, vitamin C and carotenoids) in the samples were investigated by chromatographic and spectroscopic techniques. Irradiation resulted in a general increase of quinone radical content in all of the investigated samples, as revealed by electron paramagnetic resonance spectroscopy. The fate of these radicals after storage for 3 months was also investigated. The cellulose radical was clearly observed in a few samples. Significant losses of total ascorbate were found for black pepper, cinnamon, nutmeg, oregano, and sage, whereas a significant decrease of carotenoids content was observed for cinnamon, oregano, parsley, rosemary, bird pepper, and sage. PMID- 12568552 TI - Phytochemical composition and antioxidant stability of fortified yellow passion fruit (Passiflora edulis). AB - Yellow passion fruit juice (PFJ, Passiflora edulis f. flavicarpa) is an important component of many tropical fruit beverages, but limited data exist on its antioxidant chemical composition and stability during processing and storage. PFJ fortified with ascorbic acid (450 mg/L) and sucrose (10%) was compared to a nonfortified control, and each was evaluated with and without vacuum deaeration to remove dissolved oxygen. Following pasteurization, juices were stored for 28 days at 37 degrees C to accentuate physicochemical changes. Pasteurization (85 degrees C for 30 min) resulted in minor changes to physicochemical attributes, but appreciable changes occurred during storage that resulted in termination of the study after 28 days. Oxygen control strategies proved to be ineffective for quality retention and indicated oxygen-independent reactions affecting juice color, phytochemical content, and antioxidant activity. Ascorbic acid and sucrose fortification had an overall preservation effect on total carotenoids, the former resulting in hyperchromic shifts in absorbance, indicating their chemoprotection. Pasteurization resulted in a 25% loss in l-ascorbic acid, which was completely destroyed after 14 days of storage; losses coincided with increased juice browning and formation of 5-hydroxymethylfurfural. Numerous polyphenolics were present in PFJ, and 16 of them were tentatively characterized on the basis of spectral similarities to known standards. Individually, polyphenolics increased during pasteurization, only to decline during storage at elevated temperatures. Antioxidant activity was measured in PFJ and in two subfractions (hydrophilic and lipophilic) after processing and storage, but antioxidant values were nonadditive. A significant chemical interaction affecting antioxidant capacity was found for hydrophilic juice components, but none was observed in the presence of lipophilic phytochemicals. Physicochemical attributes and overall quality of PFJ were retained following pasteurization but were significantly impacted by degradative reactions during accelerated storage. PMID- 12568553 TI - Antioxidant properties of ready-to-drink coffee brews. AB - The influence of some technological variables on the changes of the antioxidant capacity of ready-to-drink coffee brews was investigated. Results showed that, depending on the roasting degree as well as on the packaging conditions adopted, redox reactions, which can take place during storage, are responsible for significant changes in the overall antioxidant capacity of the product. In particular, the redox potential of air-packaged coffee brews, obtained from light and medium-roasted beans, showed maximum values after 2 days of storage, which corresponded to a minimum in the chain-breaking activity, while, in the case of the dark-roasted sample packaged under ordinary atmosphere, both the redox potential and the chain-breaking activity showed a maximum around 2-3 days of storage. In contrast, in the absence of oxygen, the coffee brews maintained the initial reducing properties over all the storage time, although the radical scavenging activity values changed in a way very similar to that of the air packaged sample. These results suggested that the changes in the antioxidant properties of the coffee brews may be attributed to a further development of the Maillard reaction during storage. PMID- 12568554 TI - Biodegradation of oak (Quercus alba) wood during growth of the shiitake mushroom (Lentinula edodes): a molecular approach. AB - The chemical transformations that occur during growth of the shiitake mushroom (Lentinula edodes) on oak (Quercus alba) were investigated to improve mushroom cultivation and utilization of the spent substrate. Oak logs were decayed by L. edodes over 8 years, during which time they were sampled at six intervals (30, 40, 66, 76, 77, and 101 months). Fresh and decayed oak samples were analyzed using solid-state (13)C NMR and pyrolysis-gas chromatography-mass spectrometry as well as off-line thermochemolysis with tetramethylammonium hydroxide. Degraded oak exhibited lower carbon contents and increased oxygen content compared to the control. Solid-state (13)C NMR analysis revealed that polysaccharides were the major component of both fresh and decayed oak but that L. edodes mediated the preferential loss of cellulose and xylans as compared to lignin, which remained in an altered form. Several trends point toward the degradation of lignin, including a decrease in the proportion of syringyl units as compared to guaiacyl units and a reduction in side-chain length. An increase in guaiacyl and syringyl acid-to-aldehyde ratios occurred with growth, which suggested that the fungus had caused oxidation of Calpha-Cbeta bonds. The overall effect of L. edodes on oak is similar to that of many white-rot fungi, which simultaneously degrade all cell wall components. PMID- 12568555 TI - Phytochemical stability and color retention of copigmented and processed muscadine grape juice. AB - Muscadine (Vitis rotundifolia) grape juice was assessed for color and phytochemical stability as influenced by anthocyanin copigmentation with a water soluble rosemary extract, fortification with ascorbic acid, and processing by heat or high hydrostatic pressure (HHP). The roles of polyphenolic cofactors in the presence and in the absence of ascorbic acid were assessed as a means to improve the overall processing stability of the juice. Addition of rosemary extract from 0 to 0.4% (v/v) readily formed copigment complexes with anthocyanins and resulted in concentration-dependent hyperchromic shifts from 10 to 27% that corresponded to increased antioxidant activity. The presence of ascorbic acid was generally detrimental to juice quality, especially in the presence of rosemary extract, and resulted in overall anthocyanin, ascorbic acid, and antioxidant activity losses. Although thermal and high-pressure processing methods were detrimental to juice quality, HHP resulted in greater losses after processing, likely due to action from residual oxidase enzymes. Although physicochemical attributes were enhanced by copigmentation with rosemary extract, methods to inactivate residual enzymes should be addressed prior to copigmentation to prevent degradation of anthocyanins in the presence of ascorbic acid. PMID- 12568556 TI - Drying temperature and relative humidity effects on wheat gluten film properties. AB - The mechanical and physical properties of glycerol-plasticized wheat gluten films dried at different temperatures (20, 50, and 80 degrees C) and relative humidities (35 and 70% RH) were investigated. Dispersion of wheat gluten was prepared at pH 11 in aqueous solution. Films were obtained by casting the wheat gluten suspension, followed by solvent evaporation in a temperature and relative humidity controlled chamber. Decreasing relative humidity altered most of the mechanical properties. At 35% RH, tensile strength increased when drying temperature increased. However, at 70% RH, tensile strength decreased when temperature increased. Thickness of the films decreased by increasing temperature. Hypothetical coating strength increased with increasing drying temperature at 35% RH. However, at 70% RH, a maximum value was observed at 50 degrees C. Films produced at 80 degrees C exhibited low solubility in aqueous solution. Addition of 1.5% (w/v) sodium dodecyl sulfate increased solubility of all of the films except the film dried at 50 degrees C and 70% RH. Overall, drying temperature and relative humidity affected mechanical and physical properties of the wheat gluten films. However, the effect of drying temperature was more pronounced than the effect of relative humidity. PMID- 12568557 TI - Oil goldenberry (Physalis peruviana L.). AB - Whole berries, seeds, and pulp/peel of goldenberry (Physalis peruviana L.) were compared in terms of fatty acids, lipid classes, triacylglyerols, phytosterols, fat-soluble vitamins, and beta-carotene. The total lipid contents in the whole berries, seeds, and seedless parts were 2.0, 1.8, and 0.2% (on a fresh weight basis), respectively. Linoleic acid was the dominating fatty acid followed by oleic acid as the second major fatty acid. Palmitic and stearic acids were the major saturates. In pulp/peel oil, the fatty acid profile was characterized by higher amounts of saturates, monoenes, and trienes than in whole berry and seed oils. Neutral lipids comprised >95% of total lipids in whole berry oil and seed oil, while neutral lipids separated in lower level in pulp/peel oil. Triacylglycerols were the predominant neutral lipid subclass and constituted ca. 81.6, 86.6, and 65.1% of total neutral lipids in whole berry, seed, and pulp/peel oils, respectively. Nine triacylglycerol molecular species were detected, wherein three species, C54:3, C52:2, and C54:6, were presented to the extent of approximately 91% or above. The highest level of phytosterols was estimated in pulp/peel oil that contained the highest level of unsaponifiables. In both whole berry and seed oils, campesterol and beta-sitosterol were the sterol markers, whereas Delta5-avenasterol and campesterol were the main 4-desmethylsterols in pulp/peel oil. The tocopherols level was much higher in pulp/peel oil than in whole berry and seed oils. beta- and gamma-tocopherols were the major components in whole berry and seed oils, whereas gamma- and alpha-tocopherols were the main constituents in pulp/peel oil. beta-Carotene and vitamin K(1) were also measured in markedly high levels in pulp/peel oil followed by whole berry oil and seed oil, respectively. Information provided by the present work is of importance for further chemical investigation of goldenberry oil and industrial utilization of the berries as a raw material of oils and functional foods. PMID- 12568558 TI - Determination of tea polyphenols and caffeine in tea flowers (Camellia sinensis) and their hydroxyl radical scavenging and nitric oxide suppressing effects. AB - The native occurrence of tea polyphenols, namely, (-)-epicatechin, (+)-catechin, (-)-epigallocatechin 3-gallate, (-)-epicatechin, and (-)-epicatechin 3-gallate, and caffeine in tea flowers was assessed by an isocratic HPLC procedure. The levels of total catechins and caffeine were determined in tea flowers collected from 10 different species of Camellia sinensis. The results showed the levels of total catechin ranged from 10 to 38 mg/g, whereas the level of caffeine ranged from 3 to 8 mg/g. Levels of catechins and caffeine in tea leaves and various teas were also determined and ranged from 2 to 126 mg/g and from 23 to 49 mg/g, respectively. Both tea flower and tea leaf extracts exert their strong hydroxyl radical scavenging effects in the Fenton reaction system and nitric oxide suppressing effects in LPS-induced RAW 264.7 cells. Most tea flowers contain less caffeine, but comparable amounts of total catechins, compared to tea leaves and teas. The present study demonstrates that both tea flowers and tea leaves contain appreciable amounts of catechins and caffeine. It is likely that tea flowers might be useful for making alternative tea beverages. PMID- 12568559 TI - Synthesis and characterization of dehydrogenation polymers in Gluconacetobacter xylinus cellulose and cellulose/pectin composite. AB - To mimic the lignin polymerization process, mats of bacterial cellulose and of a pectin/cellulose composite were used as a host matrix for in vitro polymerization of coniferyl alcohol. A diffusion cell was used to allow the diffusion of both hydrogen peroxide and coniferyl alcohol into the peroxidase impregnated cellulose mats through dialysis membranes. The results indicate that significant polymerization occurs within the mats. The resulting binary and ternary blends were imaged by scanning electron microscopy (SEM) and characterized by chemical means. The presence of pectin induces a better dispersion of the synthetic lignin in the cellulose network and enhances the proportion of alkyl-aryl-ether in the polymer. PMID- 12568560 TI - Large-scale preparation of the phytoalexin elicitor glucohexatose and its application as a green pesticide. AB - Large-scale preparation of the phytoalexin elicitor was achieved through a highly regio- and sterereoselective synthesis using 2,3,4,6-tetra-O-benzoyl-d glucopyranosyl trichloroacetimidate (1), 1,2:5,6-di-O-isopropylidene-alpha-d glucofuranose (2), and 6-O-acetyl-2,3,4-tri-O-benzoyl-alpha-d-glucopyranosyl trichloroacetimidate (3) as the synthons. Coupling of 1 with 2 gave the 1-->3 linked disaccharide; subsequent selective removal of 5,6-O-isopropylidene to give 5 followed by selective 6-O-glycosylation with 1 afforded the trisaccharide 6. Hydrolysis to remove the 1,2-O-isopropylidene was accompanied by ring expansion, giving 3,6-branched pyranosyl trisaccharide. Acetylation, selective 1-O deacetylation, and activation with trichloroacetonitrile gave the trisaccharide donor 7. The trisaccharide acceptor 9 was prepared from condensation of the disaccharide 5 with 3 and subsequent 6-O-deacetylation. Coupling of the trisaccharide donor 7 with the trisaccharide acceptor 9 and subsequent deprotection afforded the glucohexatose elicitor. The cost of the produced glucohexatose should be low enough to allow its applications in agriculture as a green pesticide. At a concentration of 5-10 mg/L, the resultant elicitor was used to treat growing orange trees and harvested oranges, giving very encouraging results, comparable with those obtained using commercial pesticides at a concentration of 1400 mg/L (Topsin-M) for growing trees and 900 mg/L (Tecto) for harvested oranges, respectively. Treatment of tomato leaves against Botrytis cinerea with the synthetic elicitor at a concentration of 10 mg/L gave 82% inhibition, comparable with the inhibition of 84% by Wanmeiling at a concentration of 1000 mg/L. Treatment of tea leaves also showed promising results. PMID- 12568561 TI - Synthesis and quantitative structure-activity relationships of oxanilates as chemical hybridizing agents for wheat (Triticum aestivum L.). AB - Chemical hybridizing agents (CHAs) can facilitate two-line breeding in heterosis programs of crops. Twenty-seven oxanilates having different aromatic substitutions were synthesized and screened as CHAs on two genotypes of wheat, PBW 343 and HD 2733, during two Rabi (winter) seasons, 2000-01 and 2001-02. The oxanilates prepared by thermal condensation of anilines with diethyl oxalate or by acylation with ethoxycarbonyl methanoyl chloride were sprayed at 1000 and 1500 ppm at the premeiotic stage of wheat, when the length of the emerging spike of the first node was 7-8 mm. Pollen sterility and spikelet sterility were measured in each treatment. Ethyl oxanilates 5, 6, and 25,containing 4-F, 4-Br, and 4 CF(3) aromatic substituents, respectively, induced greater than 98% spikelet sterility, the desired level, at 1500 ppm. Quantitative structure-activity relationship analysis revealed a direct relationship between F(p) and molecular mass but an inverse relationship between MR, E(S), and R in influencing the bioactivity. Several F(1) hybrids were developed using 5, and at least one showed heterosis. PMID- 12568562 TI - Influence of organic fertilizer application on pendimethalin volatilization and persistence in soil. AB - Laboratory studies were undertaken to evaluate the influence of fertilizers on pendimethalin volatilization and persistence in soil. Various organic fertilizers such as liquid humic substances and urea were used at doses of 100 L/ha or 170 kg of N/ha, respectively. Herbicide residues were determined in air, soil solution, and soil samples by GC-ECD; the recovery of pendimethalin from spiked fertilized or control samples was found to be 81-103%. Liquid humic fertilizers increased pendimethalin dissipation during the first part of the assay, although 4 months after application, herbicide levels were similar to those observed in unfertilized soil. Fertilization of soil with urea decreased, in general, pendimethalin volatilization but increased herbicide levels in soil solution and persistence in soil, with a pendimethalin half-life approximately 70% higher than that found in unfertilized soil. PMID- 12568563 TI - Environmental effects caused by olive mill wastewaters: toxicity comparison of low-molecular-weight phenol components. AB - Olive oil mill wastewaters (OMWs) show significant polluting properties due to their content of organic substances, and because of their high toxicity toward several biological systems. Wastewaters' toxicity has been attributed to their phenolic constituents. A chemical study of wastewaters from a Ligurian oil mill characterized phenolic products such as 1,2-dihydroxybenzene (catechol), derivatives of benzoic acid, phenylacetic acid, phenylethanol, and cinnamic acid. The OMWs were fractioned by ultrafiltration and reverse osmosis techniques and tested for toxicity on aquatic organisms from different trophic levels: the alga Pseudokirchneriella subcapitata (formerly known as Selenastrum capricornutum); the rotifer Brachionus calyciflorus; and two crustaceans, the cladoceran Daphnia magna and the anostracan Thamnocephalus platyurus. The fraction most toxic to the test organisms was that from reverse osmosis containing compounds of low molecular weight (<350 Da), and this was especially due to the presence of catechol and hydroxytyrosol, the most abundant components of the fraction. PMID- 12568564 TI - Gas chromatographic-olfactometric characterization of aroma compounds in two types of cashew apple nectar. AB - Cashew apple nectar is a secondary product from the production of cashew nuts and possesses an exotic tropical aroma. Aroma volatiles in pasteurized and reconstituted (from concentrate) Brazilian cashew apple nectars were determined using GC-MS and split, time-intensity GC-olfactometry (GC-O)/GC-FID. Methional, (Z)-1,5-octadien-3-one, (Z)-2-nonenal, (E,Z)-2,4-decadienal, (E,E)-2,4 decadienal, beta-damascenone, and delta-decalactone were identified for the first time in cashew apple products. These compounds plus butyric acid, ethyl 3 methylbutyrate, 2-methylbutyric acid, acetic acid, benzaldehyde, homofuraneol, (E)-2-nonenal, gamma-dodecalactone, and an unknown were the most intense aroma volatiles. Thirty-six aroma volatiles were detected in the reconstituted sample and 41 in the pasteurized sample. Thirty-four aroma active components were common to both samples. Ethyl 3-methylbutyrate and 2-methylbutyric acid were character impact compounds of cashew apple (warm, fruity, tropical, sweaty). Using GC-pFPD, 2-methyl-3-furanthiol and bis(2-methyl-3-furyl) disulfide were identified for the first time in cashew apple. Both were aroma active (meaty). PMID- 12568565 TI - Role of certain volatile thiols in the bouquet of aged champagne wines. AB - A method for the specific extraction of volatile thiols by use of p hydroxymercuribenzoate has made it possible to identify certain flavor-active volatile thiols in Champagne wines. Benzenemethanethiol, 2-furanmethanethiol, and ethyl 3-mercaptopropionate were present in these wines at concentrations considerably higher than their perception thresholds. Their concentrations increased gradually in proportion to the bottle aging period and sharply as a result of disgorging. The contribution of these volatile thiols to the empyreumatic nuances of the bouquet of aged Champagne wines was demonstrated for the first time. PMID- 12568566 TI - Quantitative determination of geosmin in red beets (Beta vulgaris L.) using headspace solid-phase microextraction. AB - An improved analytical method for the determination of geosmin in red beets was developed using headspace solid-phase microextraction (HSPME). Volatiles of beet juice were extracted in headspace for 2 h using a polydimethylsiloxane/divinylbenzene fiber, thermally desorbed from the fiber, and analyzed by gas chromatography. The HSPME method was determined to be suitable for geosmin analysis as evidenced by high relative recovery (99.2%), low relative standard deviation (7.48%), and reasonable detection limit (1 microg/kg of beet root tissue). The concentrations of geosmin in four beet cultivars ranged from 9.69 +/- 0.22 to 26.7 +/- 0.27 microg/kg, depending on cultivar. PMID- 12568567 TI - Biosynthetic origin of geosmin in red beets (Beta vulgaris L.). AB - Geosmin provides the characteristic but sometimes undesirable "earthy" flavor to red table beets. To date, it is not known whether geosmin is a byproduct of beet metabolism or synthesized by soil-borne microorganisms and taken up by the beets during maturation. Analysis of mature beet roots revealed that peels contained 6 times the amount of geosmin compared to the bodies and cores. Sterilized beet seeds were aseptically grown in a basal medium prior to analysis for the presence of geosmin. Using a headspace solid-phase microextraction (HSPME) method, the relative recovery of geosmin from beet seedling extracts was 72.0 +/- 4.2% with ( )-menthone as the internal standard. The presence of geosmin in aseptically grown beet seedlings was confirmed by gas chromatography-mass spectrometry using authentic geosmin as the standard. During aseptic growth, the concentration of geosmin in seedlings remained constant for up to 5 months but increased at 6 months. Geosmin added to the growth medium was not absorbed by the seedlings. These studies support the conclusion that red beets are capable of endogenous synthesis of geosmin. PMID- 12568568 TI - Chemical composition, plant genetic differences, and antifungal activity of the essential oil of Helichrysum italicum G. Don ssp. microphyllum (Willd) Nym. AB - The chemical composition of the essential oil of the Sardinian dwarf curry plant [Helichrysum italicum G. Don ssp. microphyllum (Willd) Nym] was studied. Genetic analysis suggested the presence of two chemotypes; morphological and chemical differences confirmed the presence of two chemotypes (A and B). The maximum yields were 0.18 and 0.04% (v/w) for flowering tops and stems, respectively. The concentrations of nerol and its esters (acetate and propionate), limonene, and linalool reach their highest values during the flowering stage both in flowers and in stems. Besides the essential oil, type B showed an interesting antifungal activity. PMID- 12568569 TI - Discovery and structure determination of a novel Maillard-derived sweetness enhancer by application of the comparative taste dilution analysis (cTDA). AB - Application of a novel screening procedure, the comparative taste dilution analysis (cTDA), on the non-solvent-extractable reaction products formed in a thermally processed aqueous solution of glucose and l-alanine led to the discovery of the presence of a sweetness-enhancing Maillard reaction product. Isolation, followed by LC-MS and 1D- and 2D-NMR measurements, and synthesis led to its unequivocal identification as N-(1-carboxyethyl)-6 (hydroxymethyl)pyridinium-3-ol inner salt. This so-called alapyridaine, although being tasteless itself, is the first nonvolatile, sweetness-enhancing Maillard reaction product reported in the literature. Depending on the pH value, the detection thresholds of sweet sugars, amino acids, and aspartame, respectively, were found to be significantly decreased when alapyridaine was present; for example, the threshold of glucose decreased by a factor of 16 in an equimolar mixture of glucose and alapyridaine. Studies on the influence of the stereochemistry on taste-enhancing activity revealed that the (+)-(S) alapyridaine is the physiologically active enantiomer, whereas the (-)-(R) enantiomer did not affect sweetness perception at all. Thermal processing of aqueous solutions of alapyridaine at 80 degrees C demonstrated a high thermal and hydrolytic stability of that sweetness enhancer; for example, more than 90 or 80% of alapyridaine was recovered when heated for 5 h at pH 7.0, 5.0, or 3.0, respectively. PMID- 12568570 TI - Characterization of three chitosanase isozymes isolated from a commercial crude porcine pepsin preparation. AB - Three chitosanases designated PSC-I, PSC-II, and PSC-III were purified from commercial pepsin preparation by sequentially applying pepstatin A-agarose affinity chromotography, DEAE-Sephacel ion-exchange chromatography, Mono Q column chromatography, and Mono P chromatofocusing. With respect to chitosan hydrolysis, the optimal pHs were 5.0, 5.0, and 4.0 for PSC-I, PSC-II, and PSC-III, respectively; optimal temperatures were 40, 40, and 30 degrees C; and the Km's were 5.2, 4.0, and 5.6 mg/mL. The molecular masses of the three isozymes were approximately 40 kDa, as estimated by both gel filtration and SDS-PAGE, and the isoelectric points were 4.9, 4.6, and 4.4, respectively, as estimated by isoelectrofocusing electrophoresis. All three chitosanase isozymes showed activity toward chitosan polymer and N,N",N' "-triacetylchitotriose oligomer. Most effectively hydrolyzed were chitosan polymers that were 68-88% deacetylated. PMID- 12568571 TI - Emulsifying properties of bovine serum albumin-galactomannan conjugates. AB - Bovine serum albumin (BSA)-galactomannan conjugates were prepared through the Maillard reaction. To 1 mol of BSA was bound 2.5-7 mol of galactomannan. Conjugates could be grouped into two fractions, on the basis of the weight average molar mass measured with a multiangle laser light-scattering detector, the main one with 220 000-250 000 Da and the other one with a very small amount of aggregates over 1 000 000 Da. Spectroscopic analysis suggested that the surface of the conjugate was covered with galactomannan and the conformation of the hydrophobic interior and the secondary structure were not significantly changed. The emulsifying activity index of the conjugates increased greatly as compared with that of BSA alone. All conjugates showed better stability than BSA, presumably due to the physical protection introduced by the viscoelastic galactomannan layer. The average particle sizes of the emulsions were similar. The interfacial properties of the BSA-galactomannan conjugates were improved, possibly due to the reduced hydrophobic interaction between the droplets and the viscoelastic interfacial properties of galactomannan. PMID- 12568572 TI - Effects of endogenous flour lipids on the quality of semisweet biscuits. AB - Fractionation and reconstitution techniques were used to study the contribution of endogenous flour lipids to the quality of semisweet (Rich Tea-type) biscuits. Biscuit flour was defatted with chloroform and baked with bakery fat but without endogenous lipid addition. Semisweet biscuits baked from defatted flour were flatter, denser, and harder and showed collapse of gas cells during baking when compared with control biscuits. Defatted flour semisweet doughs exhibited a different rheological behavior from the control samples showing higher storage and loss moduli (G' and G' ' values), that is, high viscoelasticity. Functionality was restored when total nonstarch flour lipids were added back to defatted flour. Both the polar and nonpolar lipid fractions had positive effects in restoring flour quality, but the polar lipid fraction was of greatest benefit. Both fractions were needed for complete restoration of both biscuit quality and dough rheological characteristics. PMID- 12568573 TI - Purification and characterization of amylases from small abalone (Sulculus diversicolor aquatilis). AB - Amylases II-1 and II-2 with molecular weights of 55.7 and 65 kDa, respectively, were purified to electrophoretical homogeneity from small abalone (Sulculus diversicolor aquatilis) by ammonium sulfate fractionation, Sepharose CL-6B, CM Sepharose CL-6B, and Sephacryl S-100 chromatographs. They had optimal temperatures of 45 and 50 degrees C and an optimal pH of 6.0. The purified amylases were stable at pH 5.0-8.0 and 6.0-8.0, respectively. They were completely or partially inhibited by Hg(2+), Cu(2+), Cd(2+), Zn(2+), iodoacetamide, phenylmethanesulfonyl fluoride, and N-ethylmaleimide, suggesting the existence of cysteine at their active sites. Digestion tests against various polysaccharides suggested that the purified amylases II-1 and II-2 are neoamylases which can hydrolyze both alpha-1,4 and alpha-1,6 glucosidic bonds. Amylase II-2 might be an exo- and II-1 an endo-/exo-amylase. PMID- 12568574 TI - Bacteriocins from Pediococcus pentosaceus L and S from pork meat. AB - Two strains of Pediococcus pentosaceus were isolated from refrigerated pork and found to produce antimicrobial substances that may inhibit foodborne pathogens and have potential as natural food preservatives. They were named P. pentosaceus L and S. The antimicrobial substances were purified to electrophoretical homogeneity by chloroform extraction and designated pentocins L and S with molecular masses (M) of 27 and 25 kDa, respectively. Both pentocins also had broad inhibition spectra and were thermostable. They inhibit the growth of tested spore-forming G+ and G- strains and the germination of Bacillus subtilis ATCC 10225, B. subtilis ATCC 10254, and Bacillus cereus ATCC 11778 spores. The inhibition activities decreased as the glucose in the medium decreased from 8.0 to 2.0%. PMID- 12568575 TI - Sequestering ability of butylated hydroxytoluene, propyl gallate, resveratrol, and vitamins C and E against ABTS, DPPH, and hydroxyl free radicals in chemical and biological systems. AB - The antioxidant capacity of butylated hydroxytoluene (BHT; 2,6-di-tert-butyl-p cresol), propyl gallate (3,4,5-trihydroxybenzoic acid n-propyl ester), resveratrol (trans-3,4',5-trihydroxystilbene), and vitamins C (l-ascorbic acid) and E [(+)-alpha-tocopherol] was studied in chemical and biological systems. The chemical assays evaluated the capacity of these antioxidants to sequester 2,2' azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS.) and 1,1 diphenyl-2 picrylhydrazyl (DPPH.). A new colorimetric method to determine hydroxyl radical scavenging is also described. The biological tests use the eucaryotic cells of Saccharomyces cerevisiae treated with the antioxidants in the presence of the stressing agents apomorphine, hydrogen peroxide, and paraquat dichloride (methylviologen; 1,1'-dimethyl-4,4'-bipyridinium dichloride). The results in chemical systems showed that all of the antioxidants were able to significantly inhibit the oxidation of beta-carotene by hydroxyl free radicals. The assays in yeast showed that the antioxidant activity of the tested compounds depended on the stressing agent used and the mechanism of action of the antioxidant. PMID- 12568576 TI - Effect of selenium on increasing the antioxidant activity of tea leaves harvested during the early spring tea producing season. AB - This research was to determine the effect of foliar application of selenium on increasing the antioxidant activity of tea harvested during the early spring tea producing season using a alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radical scavenging method and the linoleic acid system. The results showed that the radical scavenging ability of the tea extracts followed this order during the first 60 min: selenium-enriched tea obtained by fertilization with selenate > BHT > selenium-enriched tea obtained by fertilization with selenite > alpha tocopherol > regular tea. Se-enriched tea obtained by fertilization with selenate exhibited the highest inhibition percentage of 84.29% at 30 min. Se-enriched tea extracts provided higher hydrogen-donating capabilities than regular tea and contrasts with BHT and alpha-tocopherol at the concentration of 100 microg of solids/mL of ethanol. There was a little change in the sequence of radical scavenging ability during the later 60 min: Se-enriched tea obtained by fertilization with selenate > Se-enriched tea obtained by fertilization with selenite > BHT > regular tea > alpha-tocopherol. The individual activity of tea extracts and references measured by the linoleic acid system showed that the tea extracts, BHT, and alpha-tocopherol manifested almost the same patterns of activity as the DPPH method. Tea enriched in selenium by fertilization with selenate still exhibited the highest inhibition activity of lipid oxidation, whereas alpha-tocopherol showed the lowest inhibition. The antioxidant activity of Se-enriched green tea harvested during the early spring tea producing season is enhanced compared to regular tea. PMID- 12568577 TI - Ingestion of water-soluble soybean fiber prevents osteopenia and hypercholesterolemia induced by ovariectomy in rats. AB - This study examined the effects of feeding water-soluble soybean fiber (WSSF), a highly fermentable dietary fiber with low viscosity, on intestinal calcium (Ca) absorption, bone mineral content, and serum total cholesterol levels using female Sprague-Dawley rats (5 weeks old) with or without ovariectomy (OVX). The rats in each group were fed a WSSF (50 g/kg of diet) or WSSF-free diet for 4 weeks. Ca absorption was higher in OVX rats fed the WSSF diet than in OVX rats fed the WSSF free diet. Femoral Ca content in both sham and OVX rats fed the WSSF diet was higher than that in rats fed the WSSF-free diet. The serum total cholesterol levels were elevated after OVX compared with that in the sham rats, and this elevation was prevented by the WSSF diet. In conclusion, ingestion of WSSF prevented osteopenia and hypercholesterolemia induced by ovariectomy in rats. PMID- 12568578 TI - Influence of different mineral and Organic pesticide treatments on Cd(II), Cu(II), Pb(II), and Zn(II) contents determined by derivative potentiometric stripping analysis in Italian white and red wines. AB - This paper deals with the use of derivative potentiometric stripping analysis (dPSA) as a rapid and precise method to determine Cd(II), Cu(II), Pb(II), and Zn(II) levels in red and white wine samples from Sicily, Campania, and Tuscany and to investigate the possible connection between the content of these metals and the pesticide treatments used in vine-growing to control plant diseases and pests. dPSA allowed direct quantitation of heavy metals in acidified wines without any sample pretreatment. Mean recoveries of Cd(II), Cu(II), Pb(II), and Zn(II) ranged from 95.5 to 99.2% for white wine samples and from 96.1 to 100.0% for red wine samples. The obtained results showed that Cd(II) was not found in any sample and that Cu(II), Pb(II), and Zn(II) levels were always lower than the toxicity limits in both fungicide- and water-treated wines. Nevertheless, the contents of metals were increased in samples from organic and inorganic pesticides treatment with respect to the water-treated samples. In particular, quinoxyfen, dinocap-penconazole, and dinocap applications considerably increased Cu(II) and Zn(II) contents in white and red wines. The levels of lead were significantly raised by azoxystrobin and sulfur treatments. PMID- 12568579 TI - Metabolism of stevioside by chickens. AB - In intubation experiments (643-1168 mg per animal), most of the stevioside administered to chickens was recovered unchanged in the excreta, and only about 2% was converted into steviol. Neither stevioside nor steviol could be found in the blood. In chronic studies (667 mg of stevioside/kg of feed) with laying hens and meat-type chickens, no significant differences were found in feed uptake, weight gain, and feed conversion as the result of stevioside administration. The egg production and egg composition of laying hens were not influenced. Most of the stevioside taken up was found untransformed in the excreta, and about 21.5% or 7.3% was converted to steviol by meat-type chickens or laying hens, respectively. No stevioside or steviol could be detected in the blood or in the eggs of the different groups of animals. In anaerobic incubation experiments with chicken excreta, only a 20% conversion of stevioside into steviol was found. No harmful effects were observed in the chronic stevioside supplementation experiments nor in the intubation experiments in which very high stevioside doses were given. PMID- 12568580 TI - Quantitation of ochratoxin A in South African wines. AB - The natural occurrence of the carcinogenic mycotoxin ochratoxin A (OTA) in wines sold in local retail outlets in South Africa and Italy was investigated by HPLC analysis with fluorescence detection following cleanup by immunoaffinity column. All 24 local South African wines tested (15 white and 9 red) were found to contain detectable levels (>0.01 microg/L) of OTA, with a mean of 0.16 microg/L in the white wines and a mean of 0.24 microg/L in the red wines. Results were subsequently confirmed by LC-MS analysis using positive ion electrospray ionization with collision-induced dissociation of the protonated molecular ion [M + H](+) at m/z 404 and selected reaction monitoring of the resultant product ions [M + H - H(2)O - CO](+) at m/z 358 and [M + H - H(2)O](+) at m/z 386. Comparison with the fluorescence method gave a significant correlation (r = 0.87; p < 0.01). Although OTA contamination was present in all of the South African samples analyzed, levels were well below the suggested European Union limit of 0.5 microg/kg. The highest level found in a locally purchased wine was 0.39 microg/L in a blend of local and imported Spanish red wine. Of the eight Italian wines analyzed, only two red wines were contaminated above the suggested maximum level. PMID- 12568582 TI - Highly regioselective functionalization of aliphatic carbon-hydrogen bonds with a perbromohomoscorpionate copper(I) catalyst. AB - The complex Tp(Br3)Cu(NCMe) (1) is an excellent catalyst for the regioselective carbene transfer reaction to tertiary C-H bonds of hydrocarbons, at room temperature, using the readily available ethyl diazoacetate (EDA) as the carbene source. PMID- 12568581 TI - Direct observation of an acyl nitroso species in solution by time-resolved IR spectrocopy. AB - Benzoyl nitroside (5) was generated in solution by laser photolysis of 3,5 diphenyl-1,2,4-oxadiazole-4-oxide (4) and studied by time-resolved infrared spectroscopy. The second-order rate constants for reaction of 5 with diethylamine and 1,3-cyclohexadiene were determined to be (1.3 +/- 0.5) x 10(5) M(-1) s(-1) and (6.0 +/- 0.5) x 10(3) M(-1) s(-1), respectively. The formation of nitroxyl (HNO), a product of the reaction of 5 with diethylamine, was also observed. PMID- 12568583 TI - Fluorescence resonance energy transfer in a novel two-photon absorbing system. AB - A novel fluorescence resonance energy transfer (FRET) system containing a two photon absorbing dye and a nile red chromophore has been synthesized. Upon two photon excitation by laser at 815 nm this molecule displays efficient energy transfer from the two-photon absorbing dye to the nile red moiety, with an 8-fold increase in emission compared to the model compound. Similarly, single-photon excitation of the two-photon absorbing moiety at 405 nm results in >99% energy transfer efficiency, along with a 3.4-fold increase in nile red emission compared to direct excitation of the nile red chromophore at 540 nm. This system provides an effective way to use IR radiation to excite molecules that, by themselves, have little or no two-photon absorption. PMID- 12568585 TI - The first structurally characterized aluminum compound with two SH groups: [LAl(SH)(2)] (L = N(Ar)C(Me)CHC(Me)N(Ar), Ar = 2,6-i-Pr(2)C(6)H(3)) and the catalytic properties of the sulfur P(NMe(2))(3) system. AB - Surprisingly stable is the bis(hydrogen sulfide) of aluminum LAl(SH)(2) with two terminal arranged SH groups. The insertion of sulfur into the Al-H bonds is catalyzed by SP(NMe(2))(3). A possible mechanism is discussed. PMID- 12568584 TI - Converting the sacrificial DNA repair protein N-ada into a catalytic methyl phosphotriester repair enzyme. AB - Mutation of the active-site residue Cys38 of N-Ada converts it from a sacrificial DNA repair protein to an enzyme that uses methanethiol as an external sacrificial reagent to repair DNA methyl phosphotriesters catalytically. PMID- 12568586 TI - A neutral xenon-containing radical, HXeO. AB - We report an open-shell species containing xenon, HXeO ((2)Sigma), prepared by UV photolysis of H(2)O/Xe or N(2)O/HBr/Xe solid mixtures at 7 K and subsequent thermal mobilization of oxygen atoms at >/=30 K. The H-Xe stretching absorption of HXeO in solid Xe is at 1466.1 cm(-1), and it shifts to 1070.3 cm(-1) upon deuteration. The extensive ab initio calculations indicate that HXeO is intrinsically stable, owing to significant ionic and covalent contributions to its bonding. The formation of HXeO ((2)Sigma) radicals in these experiments suggests extensive stabilization and thermal mobility of singlet ((1)D) oxygen atoms in solid Xe and holds promises for the stability of the HKrO and HArO species. PMID- 12568587 TI - Wavelength-dependent photochemistry of Diazo Meldrum's acid and its spirocyclic isomer, Diazirino Meldrum's acid: Wolff rearrangement versus isomerization. AB - Photoreaction of diazo Meldrum's acid (1) shows a unique wavelength selectivity. At 254 nm it results in efficient (phi(254) = 0.34) Wolff rearrangement, while irradiation with 355 nm light leads to a completely different process, isomerization into corresponding cyclic alpha,alpha"-dicarbonyl diazirine 2 (phi(350) = 0.024). UV photolysis of diazirine 2 is accompanied by two competing processes: loss of nitrogen followed by the Wolff rearrangement and isomerization into diazo compound 1. Thermal decomposition of 1 leads to clean Wolff rearrangement, while heating of 2 causes quantitative conversion into diazo isomer 1. PMID- 12568588 TI - Enantioselective total synthesis of borrelidin. AB - The first total synthesis of the natural product borrelidin is described. The propionate fragment of the molecule was concisely synthesized through catalytic enantioselective reductive aldol reactions, a catalytic Negishi coupling, and a catalytic directed hydrogenation. The propionate segment was then fused to the vinyl iodide fragment through a catalytic Sonogashira coupling. Subsequent catalytic hydrostannylation and catalytic cyanation allowed access to the target structure. PMID- 12568589 TI - N,N'-dimethyl-N,N'-diarylurea anion radicals: an intramolecular reductive elimination. AB - The one-electron reduction of tertiary N,N'-dimethyl-N,N'-diarylureas (aryl = phenyl, beta-naphthyl, alpha-naphthyl), in HMPA, results in anion radicals that undergo novel intramolecular reductive elimination reactions leading to the formation of the anion radicals of the corresponding biaryls. These results are due to face to face pi-pi stacking interactions involving the two aromatic rings in the urea systems. The overlapping p(pi)() orbitals on the ipso carbons of opposing aryl groups evolve into a sigma bond leading to the formation of the biaryl anion radical. In the case of the N,N'-dimethyl-N,N'-di-2-pyrenylurea system, there is a node in the LUMO of the number 2 carbon, and the parent anion radical remains intact. PMID- 12568590 TI - A new valence tautomerism example in an electroactive ferrocene substituted triphenylmethyl radical. AB - A new molecular system combining an open-shell organic radical that acts as an acceptor group, different from an o-quinone moiety, covalently linked to a ferrocene moiety, acting as the donor group, is shown to exhibit valence tautomerism. PMID- 12568591 TI - Correlation between hydrogen bond lengths and reduction potentials in Clostridium pasteurianum rubredoxin. AB - 15N NMR hyperfine-shift data were collected for wild-type and site-specific mutant (V44I, V44A, and V44G) Clostridium pasteurianum rubredoxins in the oxidized state. Whereas most of the (15)N NMR signals did not exhibit large systematic changes upon mutation of residue 44, the signal from the backbone nitrogen of residue 44 itself (arrows) shifted by approximately 400 ppm. These shifts were used to determine the lengths of the hydrogen bond between the backbone amide of residue 44 and the side-chain sulfur of cysteine-44, which is covalently ligated to the iron of the metal center. The results, which demonstrated that this hydrogen bond is shorter in mutants with higher reduction potential, point to the importance of hydrogen bonds in modulating the reduction potential of iron-sulfur proteins. PMID- 12568592 TI - Modeling a central ligand in the nitrogenase FeMo cofactor. AB - In very recent work by Einsle et al. (Science 2002, 297, 1696), a new X-ray crystallographic structure of the FeMo cofactor of nitrogenase with a central ligand was presented. The central ligand is a light atom (N, O, or C), and Einsle et al. suggest that it is nitrogen. We present density functional calculations on the FeMo cofactor, and we investigate N, O, and C as central ligands. We show that both N and O lead to energetically stable FeMo cofactor structures, whereas C is energetically unfavorable. By comparison of bond geometries with the crystallographically determined values, we show that the central ligand is most likely nitrogen. PMID- 12568593 TI - A fluorescent sensor with high selectivity and sensitivity for potassium in water. AB - This communication describes a new optical sensor suitable for practical measurement of extracellular (serum or whole blood) potassium. The sensor responds rapidly and reversibly to changes in potassium concentrations typical of whole blood samples. No interferences from clinical concentrations of calcium or pH are observed, and the sodium interference is very minor. Excitation and emission occur in the visible light region. This new potassium sensor is currently used in the Roche OPTI CCA, a commercially available whole blood analyzer. PMID- 12568594 TI - [2,6-Mes(2)C(6)H(3)](2)Ga(+)Li[Al(OCH(CF(3))(2))(4)](2)(-) (Mes = 2,4,6 Me(3)C(6)H(2)): A compound containing a linear unsolvated two-coordinate gallium cation. AB - The title compound [2,6-Mes(2)C(2)H(3)](2)Ga(+)Li[Al(OCH(CF(3))(2))(4)](2)(-), 1, containing a linear two-coordinate gallium cation, has been obtained by metathesis reaction of [2,6-Mes(2)C(2)H(3)](2)GaCl with 2 equiv of Li[Al(OCH(CF(3))(2))(4)] in C(6)H(5)Cl solution at room temperature. Compound 1 has been characterized by (1)H, (13)C((1)H), (19)F, and (27)Al NMR spectroscopy and X-ray crystallography. Compound 1 consists of isolated [2,6 Mes(2)C(6)H(3)](2)Ga(+) cations and Li[Al(OCH(CF(3))(2))(4)](2)(-) anions. The C Ga-C angle is 175.69(7) degrees, and the Ga-C distances are 1.9130(14) and 1.9145(16) A. The title compound is remarkably stable, is only a weak Lewis acid, and polymerizes cyclohexene oxide. PMID- 12568595 TI - Enzymes do what is expected (chalcone isomerase versus chorismate mutase). AB - Madicago sativa chalcone isomerase (CI) catalyzes the isomerization of chalcone to flavanone, whereas E. coli chorismate mutase (CM) catalyzes the pericyclic rearrangement of chorismate to prephenate. Covalent intermediates are not formed in either of the enzyme-catalyzed reactions, K(M) and k(cat) are virtually the same for both enzymes, and the rate constants (k(o)) for the noncatalyzed reactions in water are also the same. This kinetic identity of both the enzymatic and the nonenzymatic reactions is not shared by a similarity in driving forces. The efficiency (DeltaG(o)() - DeltaG(cat)()) for the CI mechanism involves transition-state stabilization through general-acid catalysis and freeing of three water molecules trapped in the E.S species. The contribution to lowering DeltaG(cat)() by an increase in near attack conformer (NAC) formation in E.S as compared to S in water is not so important. In the CM reaction, the standard free energy for NAC formation in water is 8.4 kcal/mol as compared to 0.6 kcal/mol in E.S. Because the value of (DeltaG(o)() - DeltaG(cat)()) is 9 kcal/mol, the greater percentage of NACs accounts for approximately 90% of the kinetic advantage of the CM reaction. There is no discernible transition-state stabilization in the CM reaction. These results are discussed. In anthropomorphic terms, each enzyme has had to do what it must to have a biologically relevant rate of reaction. PMID- 12568596 TI - The development of a catalytic synthesis of munchnones: a simple four-component coupling approach to alpha-amino acid derivatives. AB - A new palladium-catalyzed route to prepare 1,3-oxazolium-5-oxides (i.e., Munchnones) directly from imine, carbon monoxide, and acid chloride building blocks has been developed. This provides a straightforward catalytic synthesis of Munchnones and is amenable to generating a diverse range of products by simple modification of the imine or acid chloride starting materials. Munchnones are vital synthetic intermediates to a variety of heterocyclic and peptide-based molecules. As such, this methodology has been utilized to design a new catalytic synthesis of alpha-amino acid derivatives via a one-pot coupling of imines, carbon monoxide, and acid chloride followed by alcohol. The latter represents the first reported catalytic synthesis of alpha-amino acids directly from imine and carbon monoxide building blocks. PMID- 12568597 TI - Direct coupling of benzene with olefin catalyzed by Pd(OAc)(2) combined with heteropolyoxometalate under dioxygen. AB - The oxidative coupling reaction of benzenes with alkenes was successfully achieved by the Pd(OAc)(2)/molybdovanadophosphoric acid (HPMoV)/O(2) system. For example, the reaction of benzene with ethyl acrylate by the above catalytic system in acetic acid afforded ethyl cinnamate as a major product in satisfactory yield. PMID- 12568598 TI - Coupling of internal alkynes in tp(me2)ir derivatives: selective oxidation of a noncoordinated double bond of the resulting iridacycloheptatrienes. AB - The reaction of different Tp(Me2)Ir derivatives and dimethylacetylene dicarboxylate (DMAD) allows the preparation of three different metallacycloheptatriene complexes and an unusual allyl-terminated metallacycle. The C atoms of distant C=C bonds in the metallacycles, including aromatic ones, can be converted selectively to the corresponding keto functionality under mild conditions. PMID- 12568599 TI - The addition polymerization of a p=c bond: a route to new phosphine polymers. AB - Addition polymerization, the most general method of preparation for organic polymers, has successfully been extended to P=C bonds. The polymerization of a phosphaalkene has been initiated by thermolysis or with alkyllithium reagents. The unprecedented poly(methylenephosphine)s are easily oxidized using oxygen or sulfur to give air stable macromolecules. A molecular weight (M(w)) of 35000 g/mol for the poly(methylenephosphine sulfude) was estimated by light-scattering GPC. PMID- 12568600 TI - Synthesis and aldol reactivity of o- and C-enolate complexes of nickel. AB - The often facile C-/O-tautomerization of transition metal enolates is severely hindered in the cyclic Ni complexes 1 and 2, allowing the study of their individual reactivities. At room temperature only the O-bound tautomer, 2, reacts with aldehydes, giving rise to the corresponding addition products. PMID- 12568601 TI - Oxidative heck-type reaction involving cleavage of a carbon-phosphorus bond of arylphosphonic acids. AB - Cleavage of a carbon-phosphorus bond is achieved under palladium catalysis in an oxidative Heck-type reaction which exploits arylphosphonic acids. The reaction of arylphosphonic acids with alkenes provides arylation products in good yields in the presence of TBAF with trimethylamine oxide as an oxidant. PMID- 12568602 TI - Highly conducting crystals based on single-component gold complexes with extended TTF dithiolate ligands. AB - Highly conducting crystals based on single-component gold complexes with extended TTF dithiolate ligands [Au(dmdt)(2)](0+) (1) and [Au(tmdt)(2)](0+) (2) were prepared (dmdt = dimethyltetrathiafulvalenedithiolate and tmdt = trimethylenetetrathiafulvalenedithiolate). On the basis of the synchrotron radiation powder diffraction data, the MEM electron density of 2 was successfully obtained. The conductivities of compacted powder samples of 1 and 2 at room temperature were 12 and 15 S cm(-1), respectively. Pauli-like susceptibility of 1 suggested the system to be essentially metallic at least above 50 K, while 2 showed a magnetic transition around 100 K without loss of its high conductivity. PMID- 12568603 TI - Characterization of (1)H-(1)H distances in a uniformly (2)H,(15)N-labeled SH3 domain by MAS solid-state NMR spectroscopy (section sign). AB - In this communication, we demonstrate the feasibility of obtaining long-range (1)H-(1)H distance information by MAS solid-state NMR for a microcrystalline, uniformly (2)H,(15)N-labeled sample of a SH3 domain of chicken alpha-spectrin. The experiments yield NOESY-type spectra and rely on the favorable dispersion of the (15)N chemical shifts of the protein backbone. Perdeuteration of nonexchangeable sites is employed to simplify proton spin systems and to obtain multiple structural information. Two mixing schemes, (1)H-(1)H double quantum filtered Post-C7 and (1)H spin diffusion, are implemented to obtain quantitative (1)H-(1)H distance information. Post-C7 and spin diffusion cross-peak buildup rates are discussed for initial-rate fitting and in the framework of n = 0 rotational resonance (rotor driven spin diffusion), respectively. Different deuteration schemes were tested to find conditions where short-range (1)H-(1)H interactions are truncated (e.g., between H(N) and H(alpha)), but long-range interactions are retained (e.g., between H(N) and H(N)). PMID- 12568604 TI - Ab initio evidence for slow fragmentation in RAFT polymerization. AB - High-level ab initio molecular orbital calculations on model reversible addition fragmentation transfer (RAFT) polymerization reactions indicate that the retardation effect that is widely observed for methyl acrylate polymerization in the presence of cumyl dithiobenzoate may be attributed to the slow fragmentation of the RAFT adduct radical. PMID- 12568605 TI - A new reaction manifold for the Barton radical intermediates: synthesis of N heterocyclic furanosides and pyranosides via the formation of the C(1)-C(2) bond. AB - The first radical intermediate in the thiourethane-mediated deoxygenation of an alcohol (Barton-McCombie reaction) can participate in an exo-hex-5-enyl- or exo hept-6-enyl-type radical cyclization when a suitable radical acceptor (e.g., alpha,beta-unsaturated ester, oxime ether, or hydrazone) is appropriately placed. Carbohydrate-derived imidazolyl and triazolyl thiourethanes with such acceptors, upon addition to excess of a good hydride donor (reverse addition), undergo efficient cyclization reactions to give N-heterocyclic furanosides, and, surprisingly even N-pyranosides. Depending on the acceptor, glycosides with either a C(2)()-amino or a C(2)()-carbon substituent are formed. PMID- 12568606 TI - A simple method to synthesize Dy(OH)(3) and Dy(2)O(3) nanotubes. AB - Here we report the first synthesis of Dy(OH)(3) nanotubes by facile hydrothermal treatment of bulky Dy(2)O(3) crystals. Dy(OH)(3) nanotubes were calcined to produce Dy(2)O(3) nanotubes. Ho(OH)(3) and Ho(2)O(3) nanotubes were also obtained by the same method. The growth of nanotubes occurred through a dissolution recrystallization process. PMID- 12568607 TI - On the alpha --> beta transition of carbon-coated highly oriented PVDF ultrathin film induced by melt recrystallization. AB - Poly(vinylidene fluoride) (PVDF) is one of the polymers which exhibit pronounced polymorphic crystalline forms, depending on crystallization conditions. Four different crystalline modifications, i.e., alpha, beta, gamma, and delta, have been reported so far. Among them, even though the alpha-form is the most common one, the beta-phase is the one that has attracted the widest interest due to its extensive piezo- and pyroelectric applications. During the past few decades, a substantial amount of work has been done in attempts to characterize these crystal modifications and transformations among them. It was well documented that the alpha-form PVDF can be easily obtained through melt crystallization of the PVDF at atmospheric pressure. Its beta-counterpart can, however, only be directly obtained by growth from solution, molecular epitaxy on the surface of potassium bromide, melt crystallization at high pressures, or by applying with a strong electric field. Transformation from its alpha-phase to beta-phase has been achieved by mechanical deformation, while the retransformation could be conducted through melt recrystallization at atmospheric pressure. In the present work, the recrystallization behavior of carbon-coated melt-drawn oriented PVDF ultrathin films at atmospheric pressure was studied by means of transmission electron microscopy and electron diffraction. The results indicate that through vacuum evaporating a thin carbon layer on the surface of highly oriented alpha-PVDF ultrathin film, not only has its high orientation been preserved after a complete melting and recrystallization process, but an alpha --> beta transition of PVDF has also been achieved through melt recrystallization at atmospheric pressure. This technique can be successfully used for preparing highly oriented beta-PVDF ultrathin films, especially patterned microstructures of PVDF with its highly oriented polar beta-phase and nonoriented nonpolar alpha-phase. PMID- 12568608 TI - Total synthesis of ingenol. AB - Total synthesis of ingenol, a diterpene isolated from the genus Euphorbia, was accomplished on the basis of the novel key reactions. The highly strained ingenane skeleton was constructed through an intramolecular cyclization reaction of an acetylene dicobalt complex followed by a rearrangement reaction of an epoxy alcohol. The C(3),C(4),C(5)-triol moiety was introduced by a stereoselective double dihydroxylation reaction of a diene having C(2)-C(3) and C(4)-C(5) double bonds. PMID- 12568609 TI - Designing protein dimerizers: the importance of ligand conformational equilibria. AB - In an effort to elucidate the role of ligand conformation in induced protein dimerization, we synthesized a flexible methotrexate (MTX) dimer, demonstrated its ability to selectively dimerize Escherichia coli dihydrofolate reductase (DHFR), and evaluated the factors regulating its ability to induce cooperative dimerization. Despite known entropic barriers, bis-MTX proved to possess substantial conformational stability in aqueous solution (-3.8 kcal/mol >/= DeltaG(fold) >/= -4.9 kcal/mol), exerting a dominant influence on the thermodynamics of dimerization. To dimerize DHFR, bis-MTX must shift from a folded to an extended conformation. From this conclusion, the strength of favorable protein-protein interactions in bis-MTX-E. coli DHFR dimers (-3.1 kcal/mol >/= DeltaG(c) >/= -4.2 kcal/mol), and the selectivity of dimerization for E. coli DHFR relative to mouse DHFR (>10(7)) could be determined. The crystal structure of bis-MTX in complex with E. coli DHFR confirms the feasibility of a close-packed dimerization interface and suggests a possible solution conformation for the induced protein dimers. Consequently, the secondary structure of this minimal foldamer regulates its ability to dimerize dihydrofolate reductase in solution, providing insight into the complex energy landscape of induced dimerization. PMID- 12568610 TI - Synthesis and NMR structure of p41icf, a potent inhibitor of human cathepsin L. AB - The total synthesis and structural characterization of the MHCII-associated p41 invariant chain fragment (P41icf) is described. P41icf plays a crucial role in the maturation of MHC class II molecules and antigen processing, acting as a highly selective cathepsin L inhibitor. P41icf synthesis was achieved using a combined solid-phase/solution approach. The entire molecule (65 residues, 7246 Da unprotected) was assembled in solution from fully protected peptides in the size range of 10 residues. After deprotection, oxidative folding in carefully adjusted experimental conditions led to the completely folded and functional P41icf with a disulfide pairing identical to that of native P41icf. CD, NMR, and surface plasmon resonance (SPR) were used for the structural and functional characterization of synthetic P41icf. CD thermal denaturation showed clear cooperative behavior. Tight cathepsin L binding was demonstrated by SPR. (1)H NMR spectroscopy at 800 MHz of unlabeled P41icf was used to solve the three dimensional structure of the molecule. P41icf behaves as a well-folded protein domain with a topology very close to the crystallographic cathepsin L-bound form. PMID- 12568611 TI - Improving the accuracy of NMR structures of RNA by means of conformational database potentials of mean force as assessed by complete dipolar coupling cross validation. AB - The description of the nonbonded contact terms used in simulated annealing refinement can have a major impact on nucleic acid structures generated from NMR data. Using complete dipolar coupling cross-validation, we demonstrate that substantial improvements in coordinate accuracy of NMR structures of RNA can be obtained by making use of two conformational database potentials of mean force: a nucleic acid torsion angle database potential consisting of various multidimensional torsion angle correlations; and an RNA specific base-base positioning potential that provides a simple geometric, statistically based, description of sequential and nonsequential base-base interactions. The former is based on 416 nucleic acid crystal structures solved at a resolution of 28 at baseline (trial entry). RESULTS: The rate of moderate or marked improvement in aphthae count and pain was 36% (5 of 14 subjects) in the placebo group and 65% (11 of 17 subjects) in the rebamipide group. During months 2 to 6 of treatment, aphthae count tended to increase and reached a peak at month 4 in the placebo group but decreased in the rebamipide group. Pain score decreased to the same extent in both groups for the first 3 months of treatment; however, in the fourth to sixth months of treatment, the pain score tended to increase in the placebo group but decreased in the rebamipide group. In patients with a monthly aphthae pain score >28 at baseline, pain and count scores decreased throughout the 6 months of rebamipide treatment but increased during the last 3 months of treatment in the placebo group (p < 0.01 for the between-group comparisons). CONCLUSIONS: Rebamipide is well tolerated and improves the aphthae count and pain score in BD patients. It may therefore be useful in the treatment and prevention of frequently recurrent oral aphthous ulcers (not restricted to BD). Administration of rebamipide is not cumbersome, and it does not cause any discomfort, which corticosteroid ointments for example may do; furthermore, there are no specific adverse drug reactions. Rebamipide is therefore recommended as a long-term treatment for recurrent oral aphthous ulcers. PMID- 12568633 TI - Profile summary. PMID- 12568632 TI - Effect of policosanol on carbon tetrachloride-induced acute liver damage in Sprague-Dawley rats. AB - BACKGROUND: Policosanol is a cholesterol-lowering drug purified from sugarcane (Saccharum officinarum, L.) wax. Beneficial pleiotropic effects of policosanol, such as inhibition of the susceptibility of low density lipoprotein to lipid peroxidation, have been shown. Policosanol has a good safety profile and well tolerated and, to date, no drug-related adverse effects have been demonstrated. Specifically, policosanol has not been shown to affect liver function or to increase liver enzyme levels in experimental or clinical studies. AIM: This study was conducted to determine whether policosanol prevents liver damage induced by carbon tetrachloride (CCl4) in rats, since this model has been associated with an increased rate of lipid peroxidation. METHODS: Male Sprague-Dawley rats were randomised to four experimental groups: negative controls (no CCl4 or policosanol, group 1); positive controls (CCl4 but no policosanol, group 2); policosanol 25 mg/kg (group 3) and policosanol 100 mg/kg (group 4). Acute liver injury was induced in groups 2, 3 and 4 by CCl4 suspended in olive oil and administered at a dose of 1590 mg/kg via intraperitoneal injection. Eighteen hours after CCl4 dosing, the rats were anaesthetised and their livers removed for histopathological studies. RESULTS: Policosanol 25 and 100 mg/kg dose dependently and significantly (p < 0.01) decreased the percentage of ballooned cells and hepatocytes with lipid inclusions and increased the percentage of normal hepatocytes compared with positive controls. The percentage inhibition of the occurrence of ballooned cells and hepatocytes with lipids was marked, reaching 71 and 49%, respectively, with the higher dose (100 mg/kg). The percentage of swollen hepatocytes was unchanged by policosanol compared with positive controls. No histological alterations in liver sections were found in the negative control group. Necrotic areas and inflammatory infiltrates were observed in the liver of seven of eight (87.5%) animals in the positive control group. However, only one of eight (12.5%) animals treated with policosanol 25 mg/kg and none (0%) treated with the higher dose (100 mg/kg) showed such a pattern. CONCLUSIONS: Policosanol protected against the histological changes characteristic of CCl(4)-induced hepatic injury in rats, a model of hepatotoxicity in which the process of lipid peroxidation plays a role. Further studies aimed at demonstrating the connection between such hepatoprotective and antioxidant effects of policosanol must be initiated. PMID- 12568635 TI - Cinacalcet: Amg 073, calcimimetics--Amgen/NPS Pharmaceuticals. PMID- 12568634 TI - Budesonide hydrofluoroalkane inhalation--Chiesi: budesonide HFA inhalation- Chiesi, budesonide Modulite, S 1320 HFA inhalation--Chiesi. PMID- 12568636 TI - Emtricitabine: 524W91, BW524W91, Coviracil, FTC. PMID- 12568637 TI - Estradiol-topical--Novavax: Estrasorb. PMID- 12568638 TI - Ferumoxtran-10: AMI 227, AMI 227cn, AMI 27, BMS 180549, Combidex, G 53425, Sinerem. PMID- 12568639 TI - Morphine/dextromethorphan--Endo: E 3231, Morphidex. PMID- 12568640 TI - MT 100. PMID- 12568641 TI - Selegiline-transdermal--Somerset: Emsam. PMID- 12568642 TI - Sitafloxacin: DU 6859, DU 6859A, Gracevit, sitafloxacin hydrate. PMID- 12568643 TI - Talaporfin: LS 11, LS11, ME 2906, mono-L-aspartyl chlorin e6, NP e6, NPE 6, taporfin sodium. PMID- 12568644 TI - Testosterone-buccal--Columbia: COL 1621, Striant, testosterone PHBT, testosterone progressive hydration buccal tablet. PMID- 12568645 TI - Trabectedin: ET 743, ecteinascidin 743, Yondelis. PMID- 12568646 TI - Who needs optics? PMID- 12568647 TI - The optical properties of the crystalline lens and their significance. AB - We do not yet have definitive data on the optical structure of the lens, including surface shapes and gradient refractive index distribution. Some data do exist but we need much more data to complete our knowledge of this structure. However, using current and sparse knowledge, we can use mathematical modelling to study the effects of surface shapes and gradient refractive index structure on the Gaussian and aberration properties of the lens. The equations extracted from this modelling show some interesting results, for example, the way the gradient index structure provides some of the refractive power and how this structure possibly predicts a negative spherical aberration of the lens, which is observed in real eyes. This modelling will provide a firm theoretical foundation for analysing more accurate and extensive lenticular shape and refractive index data as they become available. PMID- 12568648 TI - A survey of pediatric visual assessment by optometrists in New South Wales. AB - BACKGROUND: The importance of early vision care in the prevention and treatment of amblyopia is clear from an abundance of literature on visual development. Optometrists possess the necessary skills to detect and manage amblyogenic factors and thus have an important role to play in paediatric visual assessment and management. The present study investigates the role that optometrists in New South Wales are taking in paediatric vision care and the methods and strategies used in this role. METHODS: Questionnaires were sent to 400 optometrists practising in New South Wales; 179 completed questionnaires were returned. RESULTS AND DISCUSSION: Our findings reveal a number of associations between practice characteristics (such as location) and the management of paediatric patients. In general, practitioners in rural locations, in full-time practice or self-employed were more likely to treat patients and less likely to refer them elsewhere than those in city locations, in part-time practice or employed by others. In addition, our findings suggest the possibility of a changing pattern of collaboration between optometrists and ophthalmologists in the management of paediatric patients. PMID- 12568649 TI - Cataract surgery and changes in quality of life measures. AB - BACKGROUND: This study examines whether quality of life measured with non-disease specific instruments is altered by cataract surgery and whether the effect is modulated by the presence of ocular comorbidity or other non-ocular issues. METHODS: Thirteen patients were assessed pre- and at three months post-cataract surgery. Visual disability was measured with the Visual Disability Assessment. Quality of life was measured using two established generic instruments: the Cantril Ladder and the Profile of Mood States (POMS). Subjects were also asked to report any events that may have influenced their quality of life in the three months after surgery. RESULTS: Visual acuity and visual disability were significantly improved by surgery (p < 0.01). Cantril Ladder score was significantly improved after surgery (p = 0.04). The POMS score improved significantly after surgery, only when cases with ocular comorbidity were excluded (p = 0.04). Two cases had a large deterioration in POMS score that was attributable in one case, to the death of a partner and in the other (a patient with severe glaucoma) to depression from the realisation that there was no further potential for visual improvement. DISCUSSION: Quality of life can be measurably improved by cataract surgery. However, this is less likely if ocular comorbidity is present. The use of generic instruments for outcome studies increases the likely influence of external factors on quality of life measures. Disease specific instruments, such as the Visual Disability Assessment, should be used wherever possible in outcomes research. PMID- 12568650 TI - Practice of orthokeratology by a group of contact lens practitioners in Hong Kong. Part 2: orthokeratology lenses. AB - PURPOSE: The aim of the study was to provide information about the characteristics of three main types of orthokeratology (ortho-k) lenses used in Hong Kong and to report on their performance based on the clinical impressions of a group of ortho-k practitioners. METHOD: Twelve ortho-k practitioners were interviewed between 1 March and 30 June 2001. RESULTS: Most ortho-k lenses were ordered from three manufacturers: DreimLens, Fargo and Contex. The median maximum myopia reduction reported for DreimLens, Contex and Fargo lenses were 6.25 D, 6.00 D and 4.50 D respectively. The time to reduce myopia by up to 4.00 D could be up to three weeks for Contex and DreimLens and up to four weeks for Fargo. For reduction of myopia by up to 4.00 D, the treatment usually required only one or two lenses per eye for all three types of lenses. The incidence of lens binding and lens tightening after achieving the optimal reduction was reported to be higher with the DreimLens design. Good centration, less lens binding, relatively lower incidence of complications and lens tightening after achieving the optimal reduction were reported with the Fargo lenses. CONCLUSIONS: DreimLens tended to be more effective for myopia reduction. However, some practitioners were concerned with the aggressiveness of myopia reduction using this lens design and the higher potential for ocular complications. Selection of the lens design is dependent on various factors, in particular, practitioners need to be comfortable with the design they choose and to consider the needs of their patients and the final goal of the treatment. PMID- 12568651 TI - Ocular cicatricial pemphigoid. AB - Ocular cicatricial pemphigoid is a chronic progressive immunological disease, although acute inflammatory episodes may occur. Typically, it leads to fornix foreshortening, symblepharon and dry eye problems. In the end stage, the cornea is severely affected, resulting in serious vision loss. We report on the diagnosis and management of a patient with ocular cicatricial pemphigoid and comment on the differential diagnosis, complications and treatment options. PMID- 12568652 TI - Sub-retinal neovascular membrane in exudative macular degeneration. AB - Age-related macular degeneration is a major cause of serious vision loss. The earliest stages of age-related maculopathy may be defined by the size of the drusen present in the macula and the effects on vision. Further manifestations may include soft drusen, choroidal neovascularisation, macular haemorrhage and cicatricial or disciform degeneration of the macula. This report describes a patient with a macular haemorrhage, a choroidal neovascular membrane and serious loss of vision. In addition, the pathogenesis, diagnosis and treatment options of macular degeneration are reviewed. PMID- 12568653 TI - David Michael Cockburn OAM. Educator, entertainer, innovator, legend. PMID- 12568654 TI - Reading the fine print: correcting ametropia in infant visual acuity studies. PMID- 12568655 TI - Pre-steady-state kinetics of Bacillus licheniformis 1,3-1,4-beta-glucanase: evidence for a regulatory binding site. AB - In a previous paper, we reported the first stopped-flow experiments on a Bacillus licheniformis 1,3-1,4-beta-glucanase [Abel, Planas and Christensen (2001) Biochem. J. 357, 195-202]. It was shown that the pre-steady-state kinetics of the 1,3-1,4-beta-glucanase using the substrate 4-methylumbelliferyl 3-O-beta cellobiosyl-beta-D-glucoside may be explained by a reaction scheme involving an induced fit and the binding of two substrates as well as a second enzymic conformational change, whereas the results definitely could not be explained in terms of the simple double-displacement scheme. In the present study, we report further stopped-flow kinetic results on the glucanase using a series of low molecular-mass substrates with various leaving groups and varying chain length. The analysis of the resulting data leads to the conclusion that the free enzyme exists in two conformations, one of which binds the substrates rather strongly in a regulatory site, before any productive interactions can take place. This corresponds to an allosteric activation mechanism. With these substrates, however, the productive enzyme-substrate species are also able to change into less active or inactive forms. This may be seen as a feedback inhibitory mechanism. PMID- 12568657 TI - Nutritional genomics - "Nutrigenomics". PMID- 12568659 TI - Glucose transporters (GLUT and SGLT): expanded families of sugar transport proteins. AB - The number of known glucose transporters has expanded considerably over the past 2 years. At least three, and up to six, Na+-dependent glucose transporters (SGLT1 SGLT6; gene name SLC5A) have been identified. Similarly, thirteen members of the family of facilitative sugar transporters (GLUT1-GLUT12 and HMIT; gene name SLC2A) are now recognised. These various transporters exhibit different substrate specificities, kinetic properties and tissue expression profiles. The number of distinct gene products, together with the presence of several different transporters in certain tissues and cells (for example, GLUT1, GLUT4, GLUT5, GLUT8, GLUT12 and HMIT in white adipose tissue), indicates that glucose delivery into cells is a process of considerable complexity. PMID- 12568656 TI - Role of organic anion-transporting polypeptides, OATP-A, OATP-C and OATP-8, in the human placenta-maternal liver tandem excretory pathway for foetal bilirubin. AB - Recent functional studies have suggested that, in addition to simple diffusion, carrier-mediated transport may play an important role in foetal unconjugated bilirubin (UCB) uptake by the placenta. We have investigated the role of organic anion-transporting polypeptides (OATPs) in UCB transport by the placenta-maternal liver tandem. RNA was obtained from human liver (hL), human placenta (hPl) at term, and purified (> 80%) cytokeratin-7-positive mononucleated human trophoblast cells (hTCs). By analytical reverse transcription (RT)-PCR, agarose gel electrophoresis separation and sequencing, the mRNA of OATP-A ( SLC21A3 ) and OATP-8 ( SLC21A8 ) was identified in hL, hPl and hTCs, whereas that of OATP-C ( SLC21A6 ) was detectable only in hL. Real-time quantitative RT-PCR revealed that in hL the abundance of mRNA was OATP-8 > OATP-C >> OATP-A, whereas in hPl and hTCs this was OATP-8 >> OATP-A >> OATP-C. Expression levels for these OATPs were hL >> hTCs > hPl. Injection of mRNA of OATP-A, OATP-C or OATP-8 or RNA from hL, hPl or hTCs into Xenopus laevis oocytes conferred on them the ability to take up [(3)H]17 beta-D-glucuronosyl oestradiol ([(3)H]E(2)17 beta G) and [(3)H]UCB, although in the case of OATP-A mRNA, the induced uptake of [(3)H]UCB was very low. Cis -inhibition of [(3)H]E(2)17 beta G and [(3)H]UCB uptake by both unlabelled E(2)17 beta G and UCB was found in all cases. The affinity and efficiency of [(3)H]UCB transport was OATP-C > OATP-8. Kinetic parameters for [(3)H]UCB uptake induced by RNA from hTCs resembled most closely those of OATP-8. In conclusion, our results suggest that OATP-8 may play a major role in the carrier-mediated uptake of foetal UCB by the placental trophoblast, whereas both OATP-8 and OATP-C may substantially contribute to UCB uptake by adult hepatocytes. PMID- 12568660 TI - Effects of fish oil- and olive oil-rich diets on iron metabolism and oxidative stress in the rat. AB - The objective of the present study was to examine the effects of fish oil (FO)- and olive oil (OO)-rich diets on Fe metabolism and oxidative stress. Rats were fed for 16 weeks with diets containing 50 g lipids/kg; either OO, maize oil (MO) or FO. OO or MO diets contained a standard amount (100 mg/kg) of all-rac-alpha tocopheryl acetate. FO diets were supplemented with 0, 100 or 200 mg all-rac alpha-tocopheryl acetate/kg (FO-0, FO-1 or FO-2 diets, respectively). At the end of the feeding period, we measured non-haem Fe stores in liver and spleen, and erythrocyte and reticulocyte count. We also determined antioxidants and products derived from lipid peroxidation in plasma and erythrocytes. Our results showed reduced non-haem Fe stores in rats fed any of the FO diets. Reticulocyte percentage was higher in the rats fed FO-0 and FO-1. Plasma alpha-tocopherol was very low in rats fed the FO-0 diet. Rats fed the FO-1 and FO-2 diets showed higher alpha-tocopherol in plasma than the FO-0 group but lower than the MO or OO groups. We did not observe such differences in the alpha-tocopherol content in erythrocyte membranes. Superoxide dismutase and glutathione peroxidase activities were lower in the erythrocytes of rats fed the FO-0 diet. The products derived from lipid peroxidation were also higher in the FO groups. The administration of FO-rich diets increased lipid peroxidation and affected Fe metabolism. On the other hand, the OO-rich diet did not increase oxidative stress and did not alter Fe metabolism. Based on these results, we conclude that FO supplementation should be advised carefully. PMID- 12568661 TI - Relationship between tissue lipid peroxidation and peroxidizability index after alpha-linolenic, eicosapentaenoic, or docosahexaenoic acid intake in rats. AB - In a previous study, we found that the extent of dietary n-3 docosahexaenoic acid (DHA)-stimulated tissue lipid peroxidation was less than expected from the relative peroxidizability index of the total tissue lipids in rats with adequate vitamin E nutritional status. This suppression of lipid peroxidation was especially prominent in the liver. To elucidate whether this phenomenon was unique to DHA, we compared the peroxidation effects of n-3 alpha-linolenic acid (alpha-LN) and n-3 eicosapentaeonic acid (EPA) with those of DHA in rats. Either alpha-LN (8.6 % of total energy), EPA (8.2 %), or DHA (8.0 %) and one of two levels of dietary vitamin E (7.5 and 54 mg/kg diet) were fed to rats for 22 d. Levels of conjugated diene, chemiluminescence emission and thiobarbituric acid (TBA)-reactive substance in the liver, kidney, and testis were determined as indicators of lipid peroxidation. In rats fed the DHA diet deficient in vitamin E (7.5 mg/kg diet), TBA values in the liver, kidney, and testis correlated well with the tissues' relative peroxidizability indices. In rats fed the alpha-LN diet with an adequate level of vitamin E (54 mg/kg diet), a close association between relative peroxidizability indices and lipid peroxide levels was observed in all the tissues analysed. However, in rats fed either the EPA diet or the DHA diet with an adequate level of vitamin E, the extent of lipid peroxidation in each tissue was less than expected from the relative peroxidizability index. This suppression was particularly marked in the liver. We concluded that suppression of lipid peroxidation below the relative peroxidizability index was not unique to DHA, but was also seen with EPA, which has five double bonds, in rats with adequate vitamin E nutritional status, but not with alpha-LN, which has three double bonds. PMID- 12568662 TI - Whole-body protein turnover of a carnivore, Felis silvestris catus. AB - The cat (Felis silvestris catus) has a higher dietary protein requirement than omnivores and herbivores, thought to be due to metabolic inflexibility. An aspect of metabolic flexibility was examined with studies of whole-body protein turnover at two levels of dietary protein energy, moderate protein (MP; 20 %) and high protein (HP; 70 %), in five adult cats in a crossover design. Following a 14 d pre-feed period, a single intravenous dose of [15N]glycine was administered and cumulative excretion of the isotope in urine and faeces determined over 48 h. N flux increased (P<0.005) with dietary protein, being 56 (se 5) mmol N/kg body weight (BW) per d for cats fed the MP diet and 146 (se 8) mmol N/kg BW per d for cats fed the HP diet. Protein synthesis was higher (P<0.05) on the HP diet (75 (se 10) mmol N/kg BW per d; 6.6 (se 1) g protein/kg BW per d) than the MP diet (38 (se 5) mmol N/kg BW per d; 3.4 (se 0.4) g protein/kg BW per d). Protein breakdown was higher (P<0.05) on the HP diet (72 (se 8) mmol N/kg BW per d; 6.3 (se 0.7) g protein/kg BW per d) than the MP diet (44 (se 3) mmol N/kg BW per d; 3.9 (se 0.3) g protein/kg BW per d). Compared with other species the rate of whole-body protein synthesis in the well-nourished cat (9.7 (se 1.3) g protein/kg BW0.75 per d) is at the lower end of the range. These results show that feline protein turnover adapts to dietary protein as has been shown in other species and demonstrates metabolic flexibility. Further work is required to determine exactly why cats have such a high protein requirement. PMID- 12568663 TI - Intraruminal propionate supplementation modifies hindlimb energy metabolism without changing the splanchnic release of glucose in growing lambs. AB - The influence of propionate supplementation on the splanchnic metabolism of energy-yielding nutrients and the supply of glucose to the hindlimb was investigated in growing lambs. Six rumen-cannulated and multicatheterized lambs (32.2 kg), fed frozen rye grass at 690 kJ metabolizable energy intake/d per kg body weight0.75, were infused intraruminally with a salt solution (control) or with propionate solutions at 0.55 mol/d (P1) or 0.98 mol/d (P2) according to a replicated Latin square design. In the rumen fluid, supplementation decreased the acetate:propionate molar ratio from 2.36:1 to 1.37:1, without modifying the ruminal concentrations of acetate and NH4. As a result, the portal appearance of propionate increased by 51 and 72 % with P1 and P2, respectively, and that of l lactate doubled. Across the liver, net extraction of propionate increased by 47 and 67 % with P1 and P2, respectively. However, the net hepatic production of glucose remained unchanged, probably as the result of a substantial rise in insulin secretion and its hepatic extraction. Overall, the net splanchnic release of acetate, glucose and butyrate was not modified while that of l-lactate increased. Despite this, the net uptake of acetate, glucose, l-lactate and non esterified fatty acids by the hindlimb increased. Propionate probably enhanced the storage of energy-yielding nutrients in the hindlimb, despite their unchanged release by the splanchnic tissues and the unmodified insulinaemia. Regulatory mechanisms are not clear. PMID- 12568665 TI - Determination of each neutral oligosaccharide in the milk of Japanese women during the course of lactation. AB - Using reverse-phase HPLC after pyridylamination, we quantified the concentrations of major neutral oligosaccharides in the milk of sixteen Japanese women collected at 4, 10, 30 and 100 d postpartum. In colostrum and mature milk (30 d lactation), lacto-N-fucopentaose (LNFP) I was the most abundant oligosaccharide, followed by 2'-fucosyllactose (2'-FL) + lacto-N-difucotetraose (LNDFT), LNFP II + lacto-N difucohexaose II (LNDFH II), and 3-fucosyllactose (3-FL). Together these accounted for 73 % of the total weight of neutral oligosaccharides in colostrum and mature milk. Changes in concentration occurred during the course of lactation. LNFP I and 2'-FL + LNDFT increased from 4 to 10 d postpartum, and then declined by 100 d. LNFP II + LNDFH II steadily increased during the first 30 d and then declined. In contrast, 3-FL increased steadily throughout the entire 100 d of study. Large differences were observed between our data and previously published data in Italian women, in terms of both the concentration and temporal changes of each oligosaccharide. These differences may be caused by different assay methodology, although racial differences cannot be ruled out. PMID- 12568664 TI - Metabolism and requirements for calcium and phosphorus in the fast-growing chicken as affected by age. AB - Three series of experiments were conducted with fast-growing chickens in order: to evaluate the effects of dietary Ca and P on cholecalciferol metabolism and expression; to determine dietary Ca requirements; to determine dietary P requirements. The results of the first series confirmed previous results on the effects of dietary Ca and P on some variables of vitamin D metabolism and expression, Ca homeostasis and P metabolism in the young chicken (1- to 21-d old), and extended them to older birds (22- to 43-d-old). The bone formation rate and the duodenal calbindin content were maintained at high levels until the age of 43 d. Dietary Ca or P restriction increased duodenal calbindin and decreased bone ash in both 22- and 43-d-old chickens, but the effect on bone ash was less pronounced in the 43-d-old birds than in the younger ones. These results suggest that: (a) the capabilities for adaptation to dietary Ca and P restriction remain high during the whole growing period; (b) the growing broilers express a high adaptive capability even when the diet contains the recommended Ca and P contents. The results of the second and third series of experiments suggest that: (c) unlike the Ca requirements of the 1- to 22-d-old chick, P requirements for growth and bone ash are similar, and are as high in the older chicks as in the younger ones (7.4-8.3 g P/kg or 4.8-5.7 g non-phytate P/kg diet); (d) although growth and bone ash in the 29- to 43-d-old chickens appear to be less sensitive to dietary Ca content, within a range close to the calculated P requirement, 10 g Ca/kg diet appears to be required for best tibia mineralization, and to a lesser extent for better growth at this age. PMID- 12568666 TI - Possible effects of pollock and herring on the growth and reproductive success of Steller sea lions (Eumetopias jubatus): insights from feeding experiments using an alternative animal model, Rattus norvegicus. AB - The decline of Steller sea lions (Eumetopias jubatus) in the Gulf of Alaska appears to have been associated with a switch of diet from one dominated by fatty forage fishes (such as herring; Clupea pallasi) to one dominated by low-fat fish (such as pollock; Theragra chalcogramma). Observations made during the decline include reduced body size of sea lions, low pregnancy rates, and high mortality. We used the general mammalian model, the laboratory rat (Rattus norvegicus), to test whether changing the quality of prey consumed could cause changes in size and reproductive performance. Five groups of twelve female, weanling rats were fed diets composed of herring (H), pollock (P), pollock supplemented with herring oil (PH), pollock supplemented with pollock oil (PP), or a semi-purified diet (ICN). Mean body weights were greatest for H, followed by PH, P, PP and finally ICN, although ICN was the only group significantly different from the others (P<0.05). Food intakes before mating were 10 % higher for groups on the lower-fat diets (P and ICN), resulting in similar energy intakes in all groups. The protein efficiency ratio was highest for the H diet, slightly lower for all pollock diets, and significantly lower for ICN (P<0.05). The fetal weights for mothers fed P were significantly reduced (P<0.05). The present study shows that the energy content was a major limiting factor in the nutritional quality of pollock. When food intake was adjusted to meet energetic requirements, there were no detrimental consequences from eating pollock. However, supplementation of pollock meal with additional pollock oil may reduce growth and reproductive performance, although the reasons for this were not apparent. PMID- 12568667 TI - Plasma retinol and tocopherol levels in relation to demographic, lifestyle and nutritional factors of plant origin in Greece. AB - In the context of the Greek segment of the European Prospective Investigation into Cancer and Nutrition (EPIC) we have conducted a cross-sectional study to determine the effect of demographic, lifestyle and nutritional factors of plant origin on retinol, alpha-tocopherol and gamma-tocopherol. Study subjects were a random sample of forty-five men and sixty-eight women, aged 30-82 years, from the Greek EPIC cohort of 27953 volunteers. Analyses were done using linear regression models with the vitamin blood levels as dependent variables, and BMI, the intake of selected food items, total energy intake and a set of demographic factors as independent variables. Women had significantly lower plasma retinol levels (P<0.006) than men. Both alpha- and gamma-tocopherol levels increased by approximately 5 % for every additional 5 years of age. Among dietary factors, intake of non-fruity vegetables was associated with increase, while intake of added lipids other than olive oil with decrease, of plasma retinol levels. BMI was positively associated with plasma gamma-tocopherol only, while none of the foods or food groups investigated significantly affected the plasma levels of either of the two tocopherols. These findings need to be confirmed before attempts are made to use them in the context of explanatory mechanistic processes. PMID- 12568670 TI - Food sources of nutrients in the diet of Spanish children: the Four Provinces Study. AB - The aim of the present study was to assess the principal food sources of energy and nutrients among Spanish children. We used a cross-sectional study design, based on results obtained from a food-frequency questionnaire. The sample included 1112 children, aged 6-7 years, from Cadiz, Madrid, Orense and Murcia, Spain. Children were selected through random cluster-sampling in schools. We analysed the percentage contributed by each food item to total energy and nutrient intake. The most important food sources were: white bread in the case of carbohydrate (13.4 %); olive oil in the case of total lipids (18.3 %) and monounsaturated fatty acids (29.2 %); whole milk in the case of protein (10.2 %) and saturated fatty acids (14.9 %); chips (French fried potatoes) in the case of polyunsaturated fatty acids (30.4 %). The greatest proportion of Na, consumed in excess, came from salt added to meals. Ham ranked second as a source of saturated fats. Fruits and green leafy vegetables proved to have great relevance as sources of fibre and vitamins, though with regard to the latter, it was observed that fortified foods (breakfast cereals, dairy products, fruit juices, etc.) had come to play a relevant role in many cases. In conclusion, the nutritional profile of Spanish school-aged children aged 6-7 years could be improved by nutritional policies targeted at limiting their consumption of ham (cured or cooked) and of salt added to meals, replacing whole milk with semi-skimmed milk, encouraging the consumption of products rich in complex carbohydrates already present in children's diets (bread, pasta, rice) and promoting less fatty ways of cooking food. PMID- 12568669 TI - Effects of feed intake on composition of sheep rumen contents and their microbial population size. AB - The present study was conducted to determine the effect of feed intake on the composition of the rumen contents of sheep and on their bacterial densities. Whole rumen contents were sampled after a period of continuous inter-rumen infusion of 15NH3 from four rumen-cannulated wethers successively fed on a hay concentrate diet (2:1, w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight0.75. Total weight and chemical composition of rumen contents, as well as the distribution by size and chemical composition of particles, were determined. The populations of bacteria associated with the liquid (liquid associated bacteria, LAB) and solid (solid-associated bacteria, SAB) fractions of rumen digesta and the distribution of SAB according to feed particle size were also examined. The greater feed intake caused an increase in the mass of the rumen contents, while its chemical composition did not change, except for a higher content of organic matter (P=0.023). The distribution of feed particles by size was similar at both levels of intake. The concentrations of neutral- and acid-detergent fibre in feed particles decreased and those of total, dietary, and microbial N increased, both with a quadratic response (P=0.001), as particle size decreased. The proportion of LAB in the microbial biomass of rumen digesta reached only 8.0 %. This proportion and the density of LAB were unaffected by the level of feed intake, whereas an apparent reduction (10.4 %) occurred with the SAB biomass in whole rumen contents. A systematic, but not significant, reduction (mean value 11.9 %) in the level of microbial colonisation in the different particle fractions with the increase of feed intake was also observed. PMID- 12568668 TI - Carbohydrate-energy restriction may protect the rat brain against oxidative damage and improve physical performance. AB - Chronic energy restriction, alpha-tocopherol supplementation and their interaction with exhaustive exercise were investigated. Eleven-week-old male Wistar rats (n 6x 10) were fed either a control (C), a 30 % carbohydrate-energy restricted control (R) or an alpha-tocopherol-supplemented (S) diet for 5 months. The animals in each diet were divided into exercised (E) and non-exercised (NE) groups. Before killing, the exercised rats were required to run to exhaustion (39 (se 6), 69 (se 11) and 18 (se 2) min for the C, R and S groups, respectively). Lipid peroxidation (thiobarbituric acid-reactive substances; TBARS), protein damage (reactive carbonyls) and alpha-tocopherol were determined in gastrocnemius, liver, brain and/or plasma. There was no difference in lipid peroxidation between the R and C groups, but in liver and muscle peroxidation appeared significantly lower in the S than the other two diets. TBARS in the brain were similar in all groups. On the other hand, reactive carbonyls showed that both the R and S diets reduced protein damage in the brain, while exhaustive exercise increased it. For liver and muscle, however, reactive carbonyl levels were similar in all groups. alpha-Tocopherol supplementation increased the vitamin concentrations in liver, muscle and plasma, but exercise decreased them in plasma and brain. Carbohydrate-energy restriction increased (P=0.0025) resistance to exhaustive exercise considerably without depleting stores of alpha tocopherol or exacerbating oxidative damage in monitored tissues. It is concluded that while exhaustive exercise promotes a tissue-specific oxidative damage detectable only in brain proteins, both experimental diets tended to ameliorate this condition. PMID- 12568671 TI - Redistribution of abdominal fat after a period of food restriction in rats is related to the type of dietary fat. AB - The aim of the present experiment was to test the hypothesis that during refeeding a redistribution of intra-abdominal fat takes place and that both the recovery of weight and the redistribution of intra-abdominal fat are related to the type of dietary fat. The experimental study was carried out using male Sprague-Dawley rats. Three groups of animals were fed diets with three different fatty acid profiles. Each group contained two branches, one fed normally and the other fed initially with a 50 % energy reduction followed by refeeding ad libitum with the same isoenergetic diet as the control branch, giving a total of six treatments. Measurements were made of the final and incremental weight of the rat, weight of the intra-abdominal adipose tissue (total intra-abdominal, epididymal, omental and retroperitoneal adipose tissue weight), and feed efficacy (weight increment/metabolizable energy intake). Carcass, epididymal, omental, and muscle lipid contents, carcass protein and energy density were also measured. The results revealed that diets rich in fish oil or olive oil increase catch-up growth more than diets rich in saturated fats. During refeeding the lipid content in the adipose tissue increases while that of muscle tissue decreases. A diet rich in saturated fats induces a relative increase in the amount of intra abdominal adipose tissue. The lipid content in adipose and muscle tissues and the distribution of intra-abdominal fat can all be modified by the type of dietary fat. PMID- 12568672 TI - Ewe-lamb bonding behaviours at birth are affected by maternal undernutrition in pregnancy. AB - Maternal undernutrition in pregnancy results in low birth-weights and impaired postnatal survival in sheep. Largely anecdotal evidence suggests that the expression of appropriate maternal and neonate behaviours may also be disrupted by undernutrition. In the present study, we investigated the effect of a moderate (35 %) reduction in ewe nutritional intake in pregnancy on the expression of ewe lamb bonding behaviours in primiparous Scottish Blackface ewes. Low-intake (L) ewes had significantly higher plasma progesterone than high-intake (H) ewes from mid-gestation onwards (e.g. plasma progesterone at 20 weeks (ng/ml): H 15.72, L 22.38, sed 1.80, P<0.001), and a lower oestradiol: progesterone value than H ewes at delivery (H 0.46, L 0.35, sed 0.05, P<0.05). Lamb birth-weight was reduced in the L lambs compared with H lambs (mean body weight (kg): H 3.31, L 3.00, sed 0.14, P<0.05), but the incidence of malpresentation at delivery was greater in L lambs. L ewes spent significantly less time licking their lambs than H ewes after delivery (time grooming in 2 h after birth (%): H 56.12, L 48.17, sed 2.639, P<0.01) and were more aggressive towards the lambs. Lamb behaviours were not directly affected by maternal nutritional treatment, but lamb birth-weight had a significant effect on neonatal developmental progress. Low-birth-weight lambs were slower than heavier lambs to stand and sucked less frequently. In tests of maternal attachment to the lamb, H ewes received higher scores than L ewes at both 24 h after birth (ewes receiving high scores (%): H 41.3, L 21.4, P<0.05) and at 3 d postnatal. We conclude that even a moderate level of undernutrition impairs the attachment between ewes and lambs by affecting maternal behaviours expressed at birth. In addition, the results suggest that levels of nutrition resulting in a decrease in birth weight will affect neonatal lamb behavioural progress. PMID- 12568673 TI - Anthropometric measurements from a cross-sectional survey of Irish free-living elderly subjects with smoothed centile curves. AB - Anthropometric screening has been recommended for the detection of undernutrition as it is simple, inexpensive and non-invasive. However, a recent study estimating the prevalence of undernutrition on admission to hospital in Dublin, Republic of Ireland, highlighted that the anthropometric reference data currently available in the UK and Republic of Ireland are inadequate to accurately determine nutritional status. In order to provide current anthropometric data, we carried out a cross-sectional study of 874 free-living, apparently healthy Irish-born elderly individuals aged over 65 years. Height, weight, triceps skinfold thickness, mid-arm and calf circumference were measured, values for BMI, mid-arm muscle circumference and arm muscle area were calculated and smoothed centile data derived for each variable. One-third of these elderly individuals had a BMI between 20-25 kg/m2, approximately two-thirds (68.5 % of males and 61 % of females) were classified as overweight or obese, almost one-fifth having a BMI over 30 kg/m2 (17 % of men and 20 % of women). Very few were underweight, only 3 % having a BMI below 20 kg/m2. Height, weight, BMI and muscle reserves decreased with increasing age. The reduction in muscle size was associated with lower handgrip strength. Fat reserves declined with age in females only. Just over half of elderly Irish women reported participating in active leisure of 20 min duration four or more times/week, although 13 % reported having no involvement in active leisure. These data for the Irish elderly extend the data generated from a recent countrywide survey of Irish adults aged 18-64 years, thus providing suitable reference standards for nutritional assessment of elderly Irish individuals. PMID- 12568698 TI - [Pneumonia in regional hospitals]. PMID- 12568697 TI - [When to admit a patient with community-acquired pneumonia]. PMID- 12568699 TI - [Diagnosis and treatment of obstructive sleep apnea-hypopnea syndrome: the situation in Navarra (Spain)]. AB - OBJECTIVE: To assess the diagnosis and treatment of obstructive sleep apnea hypopnea syndrome (OSAHS) in Navarra (Spain) and to know the characteristics of patients treated with nasal continuous positive airway pressure (n-CPAP) in our community. METHOD: All patients receiving nasal n-CPAP from the public health service of Navarra in July 2000 were enrolled. A nurse visited each patient and filled in a form with epidemiological data, and patient and clinical characteristics. The nurse then measured CO in expired air and the n-CPAP pressure used and recorded the hour counter reading on the n-CPAP device. Between one and two months later and without prior warning, the patient was once again contacted and a second visit was made. The counter was read again and expired CO was measured. RESULTS: The prevalence of treatment with n-CPAP in Navarra was 125/100,000 inhabitants, and a gradual increase in annual prescriptions was observed. Home visits were made to 535 patients (80.7% of the total). Pneumologists wrote 83.5% of the prescriptions. n-CPAP was used a mean 6.1 hours/day, and the numbers of declared and real hours of use were statistically correlated. The mean apnea-hypopnea index was 51.5, although results were not available for 4.2% of the patients. The treatment was well tolerated according to 83.7% of the patients and 24.8% had been following it for over four years. The pathology most often associated with OSAHS was obesity (73.4% of the patients), followed by high blood pressure (40%); 299 patients (55.8%) had two or more associated diseases and 54 (9.7%) had none. Conventional polysomnography was the diagnostic method used in 45.9% of the cases, and respiratory polygraphs were available for 49.7%. Pressure was adjusted with the aid of polysomnography for 19.4% patients, whereas 32.1% had undergone no testing. Side effects of treatment were reported by 50.8%. CONCLUSIONS: a) The prevalence of n-CPAP treatment in Navarra is high and prescriptions are increasing; b) patient tolerance of and compliance with therapy are good, c) polysomnographs or respiratory polygraphs are available for diagnosis of OSAHS in most cases but the n-CPAP pressure level is adjusted without the recommended studies for many patients. PMID- 12568700 TI - [Characteristics of lung cancer in patients exposed to silica at work. A comparison of exposed and non-exposed individuals]. AB - INTRODUCTION: In 1977 silica was listed as a group 1 carcinogen (demonstrated in humans) by the International Agency for Research on Cancer. However, conflicting results from various studies have kept debate alive as to its carcinogenic capacity. The interest of this debate lies in the large number of workers exposed to silica. OBJECTIVE: To analyze the differential characteristics of lung cancer among silica exposed and silica non-exposed individuals, to identify indirectly a possible carcinogenic effect of silica. METHODS: For all males with a diagnosis of lung cancer over a period of 22 consecutive months at the National Silicosis Institute (Oviedo, Spain), we recorded work history, age, smoking habits, spirometry, the presence of pneumoconiosis and histology. RESULTS: Comparing the results for silica exposed and non-exposed individuals, we found significant differences for age (63.7 8.8 and 66.7 8.6 y, respectively; p < 0.05), smoking (44.1 22.4 and 48.9 22.3 packs/year; p < 0.05) and a Tiffeneau index under 70% (in 78% of exposed vs. 55% of non-exposed patients; p < 0.05). No significant differences in histology, signs and symptoms or radiographs were observed between the two groups. In the multifactorial analysis in function of age of onset, exposure to silica continued to be an independent predictor. CONCLUSION: Our results show that the clinical and radiological signs and histology of patients exposed or non-exposed to silica did not differ. However, lung cancer appears earlier among individuals with work-related exposure to silica and lower rates of tobacco smoking, suggesting some carcinogenic effect for silica. PMID- 12568701 TI - [Tuberculin test responses of tuberculosis patients]. AB - OBJECTIVE: To determine the response of tuberculosis patients to tuberculin skin tests. METHOD: The results of skin tests used for initial assessment of tuberculosis patients in Catalonia were reviewed (Multicenter Tuberculosis Research Project). Negative skin tests were those with indurations < 5 mm; positive tests were those with indurations measuring > or = 5 mm. Individuals were classed as having or not having risk factors for developing tuberculosis and those without risk factors were classified by age, location and extension of tuberculosis. RESULTS: Negative skin tests were seen in 1,566 patients (23%). Negative tests were more common in patients with risk factors, significantly so in those undergoing immunosuppressant therapy (50%) or with HIV infection (61%). Negative tests were less prevalent in patients with no risk factors (13%) and, among them, in children (1%), in patients between 15 and 29 years of age (10%) or in those with non-pulmonary forms (10%). Negative tests were significantly more prevalent among patients 60 to 74 years of age (27%), those over 74 (44%), and those whose disease was pulmonary and extrapulmonary (26%) or disseminated (64%). No significant differences in induration size of positive skin tests were observed for patients with and without risk factors (including HIV infection or non-infection). A normal distribution of induration size was observed in all groups. CONCLUSIONS: A negative tuberculin skin test for initial assessment should be interpreted in function of the presence or absence of risk factors, age, location or extension of tuberculosis. When a skin test is positive, the response will be similar whether or not an immunodepressant factor is present. PMID- 12568702 TI - [Quality of clinical trials published in Spain on asthma in comparison to trials in English language journals]. AB - OBJECTIVES: To evaluate the quality of clinical trials on asthma published in Spanish journals in comparison with those published in European or North American English-language journals. METHOD: Clinical trials were identified by searching MEDLINE and the Spanish Medical Index (Indice Medico Espanol), and by manual searches for trials mentioned in the bibliographies of the previously identified trials. Quality was assessed by the method proposed by Detsky and colleagues in their 1992 article in Journal of Clinical Epidemiology. RESULTS: The articles published in Spanish journals had lower mean quality scores than those in European or North American English-language journals. Trials published in Spain were less likely to give certain details of methodology, including ethics committee approval, details of randomization or patient enrollment and blinding. Among trials published in Spain, those with non-Spanish authors had lower mean quality scores. CONCLUSION: Clinical trials published in Spain on asthma treatments are of poorer quality than those published in English-language journals. Editors should consider using clinical trial quality checklists during the peer review process to raise the standards for published articles. PMID- 12568703 TI - [Low-weight syndrome associated with COPD in our setting]. AB - The high prevalence of chronic obstructive pulmonary disease (COPD) has considerable economic and health-related impact. The consequences arise largely from limitations on a patient's activity and shortened life expectancy. Low body weight has recently been implicated as a factor affecting limitations. Although the reason is not clear, weight loss appears to affect many patients (from 25 to 35% in different series), at least in Europe and North America. However, the situation is thought to be different in the Mediterranean area. OBJECTIVE: To estimate the prevalence of the low weight syndrome in patients with COPD in our area. METHODS: COPD patient characteristics monitored by our laboratory over the last two years (2000 and 2001) were reviewed. RESULTS: The prevalence of a body mass index (BMI) less than 20 kg/m2 was only 6.6% among the 3,126 patients studied. That percentage fell to 3.1% with a cutoff of 18 kg/m2. The figure was even more striking if we consider that half the patients had severe disease (FEV1 < 50% of reference). BMI was directly related to FEV1/FC and CO transfer. CONCLUSIONS: These results suggest that COPD patients in our geographic area have characteristics that distinguish them from those previously described in other countries. Specifically, the prevalence of low weight syndrome in our area appears to be lower. However, larger studies should be performed to confirm this finding. PMID- 12568704 TI - [COPD: from unjustified nihilism to reasonable optimism]. PMID- 12568705 TI - [Diagnostic criteria and treatment of pneumothorax]. PMID- 12568706 TI - [Primary T-cell pulmonary lymphoma. A case report and review of the literature]. AB - Non-Hodgkin's lymphomas rarely involve the pulmonary parenchyma. Most cases described to date involve B-cells. We report the first case of T-cell pulmonary lymphoma in the Spanish literature. The course of disease was rapid and response to treatment was poor. Such cases demonstrate the importance of open biopsy and the study of cell markers for diagnosis. T-cells lymphomas should be included in the differential diagnosis of febrile patients with pulmonary nodules. PMID- 12568707 TI - [Rendu-Osler-Weber syndrome. Pulmonary arteriovenous fistulas. A report of three cases]. AB - Hereditary hemorrhagic telangiectasia (HHT), or Rendu-Osler-Weber syndrome, is a rare disorder that is closely linked to the development of pulmonary arteriovenous malformations (PAVM). Diagnosis can be based on clinical signs such as upper respiratory tract changes or recurrent hemorrhagic events. Nevertheless, pulmonary involvement, a prognostic factor, may remain undetected. In the three HTT cases with PAVM we report, the following diagnostic information was obtained non-invasively: shunt fraction measurements (breathing 100% oxygen), echocardiographic contrast studies, and three-dimensional helical computed tomographs. Arteriography demonstrated a single PAVM in one case and the patient underwent successful coil embolization, with clinical and functional improvement. PMID- 12568708 TI - [Mediastinal lipoma and superior vena cava syndrome]. PMID- 12568709 TI - [Desmoid tumor of the chest wall]. PMID- 12568710 TI - Modulation of DNA-binding domains for sequence-specific DNA recognition. AB - Sequence-specific DNA-binding proteins use diverse mechanisms to recognize their cognate DNA sites. In addition to direct sequence-specific DNA contacts made by DNA recognition domains, extrinsic factors such as ligand binding, homo- and hetero-dimeric protein associations, and association via other transcription factors can also modulate the DNA-recognition properties of DNA-binding domains. In each case, these extrinsic factors act as molecular switches to facilitate cognate DNA recognition. In this article we review the available structural examples of how such extrinsic factors can modulate the way in which a DNA binding domain can recognize DNA. Together, these examples reveal a variety of ways in which such extrinsic factors can significantly extend the repertoire of DNA sites recognized by a given DNA-binding domain. PMID- 12568711 TI - Transcriptional control of the testis-specific histone H1t gene. AB - The mammalian testis-specific linker histone H1t is synthesized only in pachytene primary spermatocytes during spermatogenesis. In this review we summarize some of the progress made in our laboratory and in other laboratories in understanding transcriptional regulation of this gene. The gene is transcriptionally active in pachytene primary spermatocytes and is repressed in all other germinal and non germinal cell types. To place the transcriptional control of the testis-specific histone H1t gene in the proper context, we briefly review recent literature concerning mammalian linker histone genes in general and we compare and contrast these with the testis-specific histone H1t gene. PMID- 12568712 TI - Chromosome-based vectors for gene therapy. AB - Currently used vectors in human gene therapy suffer from a number of limitations with respect to safety and reproducibility. There is increasing agreement that the ideal vector for gene therapy should be completely based on chromosomal elements and behave as an independent functional unit after integration into the genome or when retained as an episome. In this review we will first discuss the chromosomal elements, such as enhancers, locus control regions, boundary elements, insulators and scaffold- or matrix-attachment regions, involved in the hierarchic regulation of mammalian gene expression and replication. These elements have been used to design vectors that behave as artificial domains when integrating into the genome. We then discuss recent progress in the use of mammalian artificial chromosomes and small circular non-viral vectors for their use as expression systems in mammalian cells. PMID- 12568713 TI - Characterization of Mcmar1, a mariner-like element with large inverted terminal repeats (ITRs) from the phytoparasitic nematode Meloidogyne chitwoodi. AB - Two copies of a new mariner-like element (MLE) presenting unusual inverted terminal repeats (ITRs), Mcmar1-1 and Mcmar1-2, were cloned and sequenced in the genome of the phytoparasitic nematode Meloidogyne chitwoodi. Although the sequence features of these Mcmar1 transposons are commonplace and link them to the mariner family, at their extremities they have large 355-pb long inverted terminal repeats that are perfectly conserved. This characteristic distinguishes them from all the other MLEs so far described that have imperfectly conserved ITRs of about 26-30 bp. In consequence, the sequenced full-length Mcmar1-1 element is 2000 bp long, and comprises an uninterrupted open reading frame (ORF) that encodes a putatively active transposase with 340 amino acid residues. The Mcmar1-2 element is a deleted form of Mcmar1-1 that contains a deletion overlapping most of the internal region of the 5'ITR and the 5' region of the transposase ORF. The presence of large ITRs in different transposons related to the Tc1-mariner super-family is discussed. PMID- 12568714 TI - The forkhead gene family of Caenorhabditis elegans. AB - The forkhead genes encode a family of transcriptional regulators with important roles in the control of animal development. The genomic sequence has revealed Caenorhabditis elegans has 15 forkhead genes of which six had been experimentally characterised previously. The remaining nine have now been investigated by RNAi and reporter gene expression pattern analysis. Two (B0286.5 and F26B1.7) have key developmental roles, four (F26D12.1, K03C7.2, F40H3.4 and C25A1.2) are expressed in, and may function in, the nervous system and three (C29F7.4, C29F7.5 and F26A1.2), which are closely related to each other, do not appear to be essential. Conservation with forkhead genes in the closely related nematode Caenorhabditis briggsae does not appear to correspond completely with demonstrated functionality in C. elegans. Comparisons of expression patterns and RNAi/mutant phenotypes for all 15 C. elegans forkhead genes reveal the potential complexity of functional interactions for this gene family. PMID- 12568715 TI - Genomic structure and characterization of the promoter region of the human NAK gene. AB - NAK has been identified as an IkappaB-kinase activating-kinase that plays an important role in NF-kappaB activation in response to several pro-inflammatory cytokines such as TNF-alpha. We describe here the genomic structure of the human NAK gene and analysis of the promoter. The gene spanned 40.5 kb and contained 21 exons with lengths ranging from 39 to 196 bp. Comparison of the phase and position of intron insertions within the human NAK gene with those within IKKalpha, IKKbeta and IKK epsilon indicated that the exon/intron organization of IKK epsilon is more highly conserved than that of IKKalpha or IKKbeta. The transcriptional start site was mapped at a position about 98 bp upstream from the translation start site by means of both an RNase protection assay and a primer extension method. Fluorescence in situ hybridization using full-length human NAK cDNA as a probe showed that the human NAK gene is localized to human chromosome 13q14.2-3, a region in which the loss of heterozygosity is associated with squamous cell carcinoma and leukemia. By using a series of deletion constructs in performing a reporter assay, a minimal 77 bp upstream of the transcriptional initiation site was shown to contribute to the major promoter activity. PMID- 12568716 TI - Sequence diversity and evolution of the malaria vaccine candidate merozoite surface protein-1 (MSP-1) of Plasmodium falciparum. AB - The merozoite surface protein-1 (MSP-1) of the malaria parasite Plasmodium falciparum is a major blood-stage antigen containing highly polymorphic tripeptide repeats in the domain known as block 2 and several non-repetitive domains that are essentially dimorphic. We have analyzed sequence variation in block 2 repeats and in non-repetitive block 17, as well as other polymorphisms within the MSP-1 gene, in clinical isolates of P. falciparum. Repeat haplotypes were defined as unique combinations of repeat motifs within block 2, whereas block 17 haplotypes were defined as unique combinations of single nucleotide replacements in this domain. A new block 17 haplotype, E-TNG-L, was found in one isolate from Vietnam. MSP-1 alleles, defined as unique combinations of haplotypes in blocks 2 and 17 and other polymorphisms within the molecule, were characterized in 60 isolates from hypoendemic Brazil and 37 isolates from mesoendemic Vietnam. Extensive diversity has been created in block 2 and elsewhere in the molecule, while maintaining significant linkage disequilibrium between polymorphisms across the non-telomeric MSP-1 locus separated by a map distance of more than 4 kb, suggesting that low meiotic recombination rates occur in both parasite populations. These results indicate a role for non-homologous recombination, such as strand-slippage mispairing during mitosis and gene conversion, in creating variation in a malarial antigen under strong diversifying selection. PMID- 12568717 TI - Comprehensive analysis of transmembrane topologies in prokaryotic genomes. AB - We analysed comprehensively transmembrane (TM) topologies of TM proteins of 50 selected prokaryotic genomes, by discriminating between TM and soluble proteins by using SOSUI, then detecting and removing signal peptides by applying 'DetecSig', and finally predicting TM topologies by employing 'ConPred'. Estimated fraction of TM proteins in proteome averaged over the 50 genomes is approximately 22%. About 13% of TM proteins were predicted to have a signal peptide, and the fraction of soluble proteins with signal peptide (secretory proteins) ranges from 8 to 18% for most majority of the genomes. The N(in)-type TM proteins with 2-, 4-, 6- and 12-tms (number of transmembrane segments) are predominant among multi-spanning TM proteins, and correspondingly, significantly higher fractions of N(out)-type TM proteins with 1-, 3-, 5- and 11-tms have a signal peptide. It is also found that the TM proteins with signal peptide tend to have a long N-tail loop. The averaged sequence length of TM proteins increases linearly with the increase of the number of TM segments, with the increasing rate of about 35 residues, suggesting a possibility that TM topologies might have been evolved by the 'internal gene duplication' mechanism. Datasets of TM topologies predicted in this study are available at http://bioinfo.si.hirosaki-u.ac.jp/ approximately TMPinGS/. PMID- 12568718 TI - Isolation of novel olfactory receptor genes in marmosets (Callithrix): insights into pseudogene formation and evidence for functional degeneracy in non-human primates. AB - Nineteen olfactory receptor (OR) genes were isolated from three OR subfamilies in two species of marmoset (Callithrix). Olfactory receptor 912-93 has high sequence similarity among marmosets and between marmosets and humans, suggesting strong conservation of function. All of the remaining seventeen OR genes identified from subfamilies 3A and 1E were pseudogenes. Following pseudogene formation, marmoset OR genes in both 1E and 3A subfamilies underwent duplications, indel events and a high rate of nucleotide substitution. These results provide a contrast to previous studies, and show that in spite of the keen olfactory sense of marmosets, they harbour many OR pseudogenes. A high rate of in vitro recombination using Pfu polymerase but not Taq polymerase was confirmed. The rapid molecular evolution of OR pseudogenes suggests that they do not provide a useful source of sequence variation for conversion to intact OR genes over evolutionary timescales. The overall pattern of OR evolution in marmosets is comparable to the 'birth-and-death' model of gene family evolution. An unbiased view on the evolutionary timing of the reduction of the functional olfactory repertoire in humans must await more data. PMID- 12568719 TI - Isolation and expression of tilapia (Oreochromis niloticus) serine 8-type GnRH coding and regulatory sequences. AB - The complete Serine 8-type gonadotropin releasing hormone (GnRH) coding sequence with a substantial 5-prime regulatory sequence (5 kb) has been isolated and characterised in Nile Tilapia (Oreochromis niloticus) from a relevant genomic library. The primary structure of the protein precursor was identified for this gene. The promoter efficacy has been tested using 0.6 kb of the GnRH promoter driving a lacZ reporter gene in both cultured spleen cells and transiently expressing zebrafish. In the cell transfection experiments, the average level of beta-galactosidase activity in transfected cells was more than 2.1 (P<0.05) times higher than the control promoter-less vector in five independent cultures indicating that the 0.6 GnRH/lacZ construct is able to express in spleen cells. In addition, the transient expression of the lacZ gene was detected in the brain of G0 zebrafish embryos (Danio rerio) 4 days after fertilisation following egg injection with the construct, which demonstrated the efficacy of the tilapia GnRH promoter. PMID- 12568720 TI - Genomic organization of human myotonic dystrophy kinase-related Cdc42-binding kinase alpha reveals multiple alternative splicing and functional diversity. AB - Myotonic dystrophy kinase-related Cdc42-binding kinase alpha (MRCKalpha) is a Cdc42/Rac interactive binding-containing serine/threonine kinase with multiple functional domains. Its roles in the regulation of peripheral actin reorganization in HeLa cells and NGF-induced neurite outgrowth in PC12 cells have been documented. Here we report the characterization of the genomic structure and alternative splicing of the human counterpart. Human MRCKalpha gene is located on chromosome 1q42.1, spanning a genomic region of 250-300 kb and is composed of 41 exons. Four exons in the internal variable region and six in the 3' end were found to undergo extensive alternative splicing, giving rise to 96 possible transcripts of different combinations. The region of the internal splice site that defines a variable region in between two functional domains of opposite regulatory effects on MRCKalpha catalytic activity, and the 3' end splice site that generates variants with differential GTPase binding activity suggest a role for these alternative splicing events in MRCKalpha regulation. PMID- 12568721 TI - Serine proteases and their homologs in the Drosophila melanogaster genome: an initial analysis of sequence conservation and phylogenetic relationships. AB - Serine proteases (SPs) and serine protease homologs (SPHs) constitute the second largest family of genes in the Drosophila melanogaster genome. Eighty-four SPs comprise less than 300 amino acid residues, and a significant portion of them are probably digestive enzymes. Some larger SPs may contain one or more regions important for protein-protein interactions, including clip domains, low-density lipoprotein receptor class A repeats, and scavenger receptor cysteine-rich domains. We identified 37 clusters of SP or SPH genes, which probably evolved from relatively recent gene duplication and sequence divergence. A majority of the SPs may be trypsin-like and activated by cleavage after a specific arginine or lysine residue. Among the 147 SPs and 57 SPHs studied, 24 SPs and 13 SPHs contain at least one regulatory clip domain. A multiple sequence alignment of the clip domains provided further information on structural conservation of these regulatory modules. Detailed sequence comparison led to an improved classification system for SPs containing clip domains. These analyses have established a framework of information about evolutionary relationships among the Drosophila SPs and SPHs, which may facilitate research on these proteins as well as homologous molecules from other invertebrate species. PMID- 12568722 TI - A simple Cre-loxP method for chromosomal N-terminal tagging of essential and non essential Schizosaccharomyces pombe genes. AB - To facilitate the N-terminal tagging of essential genes at their genomic locus and under control of their own promoters we have developed a series of novel polymerase chain reaction templates. Initially, a 1.8 kb DNA fragment is integrated upstream of the ATG of the gene of interest. This fragment encodes the tag, a loxP site, a selectable marker, an exogenous nmt1 promoter and a second loxP site. In a single homologous integration event, the gene of interest is placed under control of the thiamine regulated nmt1 promoter, allowing identification of potential integrants on the basis of phenotype. Subsequently, this integrant strain is transformed with a plasmid expressing the Cre recombinase. This results in excision of the marker and nmt1 promoter and leaves sequences encoding an in-frame tag at the N-terminus of the gene of interest under the control of its native promoter. We have created TAP-cdc22, TAP-suc22 and TAP-rad50 strains using this N-tagging system, and developed a range of vectors for introducing TAP-, (His)10HA-, (His)6Myc- and EGFP. PMID- 12568723 TI - Functional analysis of the KCS-like element of the interferon-inducible RNA specific adenosine deaminase ADAR1 promoter. AB - The ADAR1 gene encodes an RNA-specific adenosine deaminase that alters the functional activity of both viral and cellular RNAs by posttranscriptional adenosine-to-inosine RNA editing. The interferon (IFN) responsive PI promoter of the ADAR1 gene possesses an IFN-stimulated response element (ISRE) responsible for IFN-inducibility, as well as an adjacent upstream sequence, designated kinase conserved sequence-like (KCS-l) element. The KCS-l element is similar to the 15 bp KCS element so far unique to the human and mouse RNA-dependent PKR kinase gene promoters. The KCS element of the PKR kinase (PKR) promoter is essential for both basal and IFN-inducible PKR promoter activity. We have now examined the functional properties of the KCS-l element of the ADAR1 PI promoter. Electrophoretic mobility shift assays (EMSAs) detected constitutively expressed nuclear proteins that bound selectively to the ADAR1 KCS-l element. Competition EMSA and antibody supershift assays indicated that ADAR1 KCS-l-binding proteins shared some properties with PKR KCS-binding proteins. However, transient transfection analyses performed with ADAR1 PI promoter constructs possessing deletion and substitution mutant forms of the KCS-l element revealed that the ADAR1 KCS-l element was not essential for either basal or IFN-inducible promoter activity. Substitution of the ADAR1 KCS-l element with the PKR KCS element increased both basal and inducible ADAR1 PI promoter activity. These results suggest that the KCS-l element of the ADAR1 PI promoter is not functionally equivalent to the KCS element of the PKR promoter. PMID- 12568724 TI - Identification and characterization of ADAM32 with testis-predominant gene expression. AB - Members of the ADAM (A Disintegrin And Metalloprotease) family of cell surface proteins have been discovered in a variety of tissues and species. We have cloned and sequenced a mouse cDNA encoding a novel member of the ADAM family. This mouse ADAM gene was named ADAM32 after an human homologue previously sequenced but not characterized. The cDNA sequence of ADAM32 has an open reading frame which encodes 750 (mouse ADAM32) and 787 (human ADAM32) amino acids, sharing 69% homology between the two species. The deduced amino acid sequences of mouse and human ADAM32 cDNAs contain all of the domains common to ADAM family members. Comparison of amino acid sequences showed that ADAM32 is highly homologous to ADAM2 and ADAM3, sperm surface proteins required for fertilization. We found that the ADAM32 gene is expressed predominantly in the testis. Examination of transcription in pre-pubertal and adult mouse testes revealed that expression of ADAM32 begins during meiotic prophase, in pachytene spermatocytes. These results suggest a potential role for ADAM32 in sperm development or fertilization. Genome database searches revealed that the mouse and human ADAM32 genes are located on chromosomes 8 and 8q11, respectively. The ADAM32 gene covers a relatively large region of the genome (120 kb for mouse ADAM32 and 180 kb for human ADAM32) and consists of multiple small exons (at least 22 exons in mouse ADAM32 and 25 exons in human ADAM32). PMID- 12568726 TI - Genomic organization, splice products and mouse chromosomal localization of genes for transcription factor Nuclear Factor One. AB - Transcription factor Nuclear Factor One (NFI) proteins are derived from a small family of four vertebrate genes (NFIA, B, C and X), all of which produce a fair number of protein variants by alternative splicing. In order to ultimately locate RNA signal sequences around exon/intron borders for the production of regulated splice variants, we have determined the exon structure of the chicken NFIB gene as the last of the four vertebrate genes for which the gene structure was not yet elucidated. This made it possible to compile nine newly isolated and sequenced mouse NFI cDNA sequences together with all previously available ones and to deduce corresponding splicing patterns for the orthologous vertebrate genes of all four paralogous gene types. Results from the analysis of alternative splicing and of NFI gene mapping in the genome of human and mouse argue for a phylogenetic route in which the four vertebrate NFI genes result from a single duplication of a genomic segment containing two NFI intermediate genes rather than from two independent duplications of two separated single ancestor genes. PMID- 12568725 TI - Gene structure and promoter analysis of the rat BTEB2 gene. AB - Rat BTEB2 protein is a transcription factor with three zinc fingers that binds to GC box, and is expressed in the placenta, intestine, and testis. To understand mechanisms of gene expression of BTEB2, we have cloned the rat BTEB2 gene from a rat liver genomic library and determined the gene structure. The BTEB2 gene contained 4 exons. In the process of cloning of the BTEB2 gene, we cloned two pseudogenes for BTEB2, one of which was a processed gene. The upstream region of the bona fide gene was fused to a luciferase reporter gene, and the generated BTEB2-luciferase chimeric plasmid was transiently transfected into HeLa cells that expressed endogenous BTEB2 mRNA. Significant expression of luciferase activity was observed. Deletion analysis of the promoter region of the BTEB2 gene revealed that at least three regions are important for the activity. Upon investigation of cis-acting elements in the regions, the GC box, CCAAT box and NF 1 binding site were found. As binding factors, Sp1, CBFa, and NF-1 were identified to the DNA elements by gel mobility shift assays using specific antibodies. PMID- 12568727 TI - Pentamer vocabularies characterizing introns and intron-like intergenic tracts from Caenorhabditis elegans and Drosophila melanogaster. AB - Overall compositional properties at the level of bases, dinucleotides and longer oligos characterize genomes of different species. In Caenorhabditis elegans, using recurrence analysis, we recognized the existence of a long-range correlation in the oligonucleotide usage of introns and intergenic regions. Through correlation analysis, this is confirmed here to be a genome-wide property of C. elegans non-coding portions. We then investigate the possibility of extracting a typical vocabulary through statistical analysis of experimentally confirmed introns of sufficient length (>1 kb), deprived of known splice signals, the focus being on distributed lexical features rather than on localized motifs. Lexical preferences typical of introns could be exposed using principal component analysis of pentanucleotide frequency distributions, both in C. elegans and in Drosophila melanogaster. In either species, the introns' pentamer preferences are largely shared by intergenic tracts. The pentamer vocabularies extracted for the two species exhibit interesting symmetry properties and overlap in part. A more extensive investigation of the interspecies relationship at the level of oligonucleotide preferences in non-coding regions, not related by sequence similarity, might form the basis of new approaches for the study of the evolutionary behaviour of these regions. PMID- 12568728 TI - Genomic organization and promoter characterization of the murine dopamine receptor regulating factor (DRRF) gene. AB - To study the transcriptional mechanisms by which expression of the dopamine receptor regulating factor (DRRF) gene is regulated, a murine genomic clone was isolated using a DRRF cDNA as probe. A 24 kb genomic fragment which comprises 13 kb upstream of the transcription initiation site was sequenced. The promoter region lacks a TATA box and CAAT box, is rich in G+C content, and has multiple putative binding sites for the transcription factor Sp1. The DRRF gene also has consensus sequences for AP1 and AP2 binding sites. The transcriptional activity of five deletion mutants of a 1.5 kb fragment was analyzed by modulating transcription of the heterologous chloramphenicol acetyltransferase (CAT) gene in the promoterless plasmid pCAT-Basic. All mutants showed significant transcriptional activity in the murine neuroblastoma cell line NB41A3, except the construct stretching from -901 to +17. These transient expression assays suggested the presence of positive regulators between -1153 and -901 and between 118 and -93 while a negative regulator was found in the region between -901 and 118. Comparison among cell types revealed strong transcriptional activity of the DRRF promoter in neuronal NB41A3 cells and moderate activity in hepatic HepG2 and renal OK cells, but none in skeletal muscle C2C12 or glial C6 cells. These findings confirm the tissue-specific activity of the DRRF promoter and suggest that this gene shares structural and functional similarities with the dopamine receptor genes that it regulates. PMID- 12568729 TI - Characterization of the human Snrpn minimal promoter and cis elements within it. AB - Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are caused by loss of gene function of the imprinted genes including Snrpn within a 2 Mb domain on chromosome 15q11-13. Based on microdeletions in PWS and AS patients, a 4.3 sequence around Snrpn promoter/exon 1, together with a 880 bp sequence upstream to Snrpn, are believed to encompass an imprinting control center for the entire 2 Mb domain. We have previously characterized the mouse Snrpn minimal promoter and a 7 bp element (SBE) within it, which is required for its activity. Here we describe the human Snrpn minimal promoter sequence, which is comprised of a 71 bp upstream sequence and 51 bp of exon 1. The SBE, which has been shown to be critical for mouse promoter activity, is also found in the human sequence and absolutely required for promoter activity. Methylation of this element, like in the mouse, prevents the binding of a protein factor and abolishes promoter activity. In addition, the 5' end of exon 1 must contain cis elements that support promoter activity. In contrast, the 3' end of exon 1 appears to repress promoter activity. This sequence specifically binds a protein factor which presumably exerts a repressory effect on the promoter. Methylation of this sequence prevents the binding of this protein. PMID- 12568730 TI - The clinical practice impact of the Women's Health Initiative: political vs biologic correctness. PMID- 12568731 TI - Critical comments. PMID- 12568732 TI - Effect of vitamin K2 on cortical and cancellous bones in orchidectomized young rats. AB - OBJECTIVES: The purpose of the present study was to compare the effect of vitamin K(2) on cortical and cancellous bones in orchidectomized young rats. METHODS: Forty male Sprague-Dawley rats, 6 weeks of age, were randomized by stratified weight method into four groups with 10 rats in each group: baseline controls (BLC), age-matched controls (AMC), orchidectomy (ORX), and ORX+vitamin K(2) administration (K). Vitamin K(2) (menatetrenone) was administered subcutaneously twice a week at dose of 30 mg/kg each. The experimental period was 8 weeks, and cortical and cancellous bone histomorphometry was performed on the tibial shaft and the proximal tibia, respectively. RESULTS: Cortical area (Ct Ar) and cancellous bone volume (BV/TV) were significantly greater in the AMC group than in the BLC group. Ct Ar was significantly lower in the ORX group than in the AMC group, and cancellous BV/TV was also significantly lower in the ORX group than in the AMC group as a result of significantly increased eroded surface (ES/BS). Although Ct Ar in the ORX+K group did not differ significantly from that in the ORX group, cancellous BV/TV was significantly greater in the ORX+K group than in the ORX group, but still significantly lower than in the AMC group. This protective effect of vitamin K(2) on cancellous bone was attributable to normalizing increased ES/BS. CONCLUSIONS: Vitamin K(2) appears to act more strongly on cancellous bone than on cortical bone in ORX young rats. High dose vitamin K(2) could partially prevent the reduction of cancellous bone gain by normalizing raised bone resorption in ORX young rats. PMID- 12568733 TI - The effect of hormone replacement therapy on atherosclerotic severity in relation to ESR1 genotype in postmenopausal women. AB - OBJECTIVE: The atheroprotective action of estrogen is mediated by estrogen receptors (ESR) 1 and 2, expressed in atherosclerotic lesions. The effects of hormone replacement therapy (HRT) and ESR1 PvuII genotypes on atherosclerosis have not previously been studied prospectively in postmenopausal women. METHODS: We investigated the effect of HRT on the progression of atherosclerosis in a 5 year follow-up study of 88 postmenopausal women aged 45-71 years at baseline allocated into three groups based on the use of HRT. The HRT-EVP group (n=26) used sequential estradiol valerate (EV) plus progestin (P), the HRT-EV group EV alone (n=32), and a control group (n=30) was without HRT. The atherosclerosis severity score (AS) of the abdominal aorta and carotid arteries were determined by sonography and the ESR1 PvuII genotypes (P/P, P/p and p/p) by PCR. RESULTS: HRT, time and ESR1 PvuII genotype had a statistically significant or borderline significant main effect on AS during 5-year follow-up (P=0.004, P<0.001 and P=0.090, respectively), when analyzed by repeated measures analysis of variance. There was a significant genotype-by-treatment (HRT-EVP and control groups) interaction for AS (P=0.034). In response to HRT-EVP, subjects with P/P, compared with those with P/p and p/p genotypes, had a less increase in AS (1.61+/-1.14 vs. 1.71+/-1.27 vs. 2.43+/-1.27). Baseline AS as covariate in similar model does not change the significant interaction effect between HRT-EVP and control groups (P=0.036). But this effect was not found between HRT-EV and control groups. CONCLUSIONS: Our results suggest that the effect of HRT-EVP in postmenopausal women on progression of AS may be determined in part by the genotype of ESR1 PvuII. PMID- 12568734 TI - Estrogen effect on heart rate variability in hypertensive postmenopausal women. AB - Healthy postmenopausal women and hypertensive patients show an imbalance in the modulation of autonomic nervous control of the cardiovascular system, which may increase the cardiovascular risk. OBJECTIVE: To examine the heart rate variability (HRV) response to estrogen replacement therapy (ERT) and its association with changes in metabolic variables in hypertensive postmenopausal women. METHODS: A double-blind, placebo-controlled clinical trial was conducted in 30 hypertensive postmenopausal women receiving 180 mg/day of verapamil. The experimental group (n=16) received 0.625 mg OD of natural conjugated estrogens during 4 months, while control group (n=14) received a placebo. Lipids, lipoproteins, apolipoproteins, glucose and insulin were measured at 0, 2 and 4 months. HRV was determined in time and frequency domains using a 24-h Holter before and after ERT. RESULTS: Significant higher values of spectral and non spectral parameters of HRV, associated with a lower LF/HF ratio, were found at the end of 4 months of ERT. Multiple regression analysis revealed that estrogen treatment itself and changes in total cholesterol, LDL-cholesterol, glucose and waist circumference, contributed to the changes observed in indexes reflecting parasympathetic activity in time and frequency domains. CONCLUSIONS: We conclude that ERT partially improves HRV favoring increased parasympathetic drive, and that part of the effect may be mediated by changes in metabolic variables. PMID- 12568735 TI - Interleukin-1beta and interleukin-1 receptor antagonist gene polymorphism in postmenopausal women: correlation to bone mineral density and susceptibility to osteoporosis. AB - OBJECTIVE: Osteoporosis is a common disorder with a strong genetic component. Our aim was to investigate the correlations of the interleukin-1beta (IL-1beta) and interleukin-1 receptor antagonist (IL-1Ra) gene polymorphisms with bone mineral density (BMD) and their relationship to osteoporosis. METHODS: The IL-1beta (promoter and exon 5) and IL-1Ra (intron 2) gene polymorphisms were determined using polymerase chain reaction. BMD of the lumbar spine and proximal femur were measured using dual-energy X-ray absorptiometry. RESULTS: The prevalence of each genotype of the interleukin-1 related genes in the study population was: (1) 14% C/C, 71.5% C/T, and 14.5% T/T in IL-1beta promoter; (2) 95.3% E1/E1 and 4.7% E1/E2 in IL-1beta exon 5; (3) 92.4% I/I, 6.4% I/II, and 1.2% II/II in IL-1Ra intron 2. After adjustment for potential confounding factors such as age, height, weight, years since menopause, and daily calcium intake, subjects with genotype E1/E2 (n=8) in IL-1beta exon 5 had lower BMD values and a significantly greater risk for osteoporosis (OR 10.6, 95% CI 1.3-83.8) at the lumbar spine when compared with subjects with genotype E1/E1 (n=164) in IL-1beta exon 5. CONCLUSION: The Taq I IL-1beta exon 5 gene polymorphism is associated with reduced BMD and predisposes women to osteoporosis at the lumbar spine, but our results should be interpreted with caution because of the small number of subjects with the unfavorable E1/E2 genotype. PMID- 12568737 TI - Tear function changes of postmenopausal women in response to hormone replacement therapy. AB - OBJECTIVE: The goal of this study was to search the effect of hormone replacement therapy (HRT) on tear function changes in postmenopausal women. METHODS: Following initial ophtalmic evaluation and tear sample collection, the subjects were given daily 0.625 mg conjugated estrogen, and either continuous combined or cyclic 5 mg medroxyprogesterone acetate was added. Eye examination included visual acuity, slit-lamp examination, fundus examination, tonometry, Schirmer's test, and break up time (BUT) evaluation. Six months later, control examination was done and repeat tear samples were obtained. Tear immune globulin A (IgA) and lysozyme levels were measured by gel electrophoresis. The pictures of the bands were evaluated by digital image analysis with Scion Image program. RESULTS: Conjunctival vascular congestion, laxity and corneal desquamation did not change before and after HRT (P>0.05). A significant improvement was noted in meibomian gland inflammation (P=0.034). We have not observed any significant difference in burning, foreign body sensation, and tearing (P>0.05). In addition, no significant difference was noted in BUT (P=0.370) and Schirmer's test values (P=0.271). Though both lysozyme and IgA levels were elevated following the therapy, only IgA levels increased significantly (P=0.04). CONCLUSION: HRT decreased meibomian gland inflammation and increased tear lysozyme and IgA levels in postmenopausal women. PMID- 12568736 TI - A pilot study comparing the clinical effects of Jia-Wey Shiau-Yau San, a traditional Chinese herbal prescription, and a continuous combined hormone replacement therapy in postmenopausal women with climacteric symptoms. AB - OBJECTIVES: Interest in use of alternative remedies for managing menopausal symptoms is increasing exponentially during these years. Jia-Wey Shiau-Yau San (JWSYS), one of the traditional Chinese herbal prescriptions, is a famous herbal remedy used for the management of various menopausal-related symptoms. A randomized, controlled pilot study was performed to evaluate the clinical effects of JWSYS compared with those of a continuous combined hormone replacement therapy, Premelle, on quality of life in non-hysterectomized postmenopausal women. METHODS: The present trial compared the effect of a l6-week treatment with JWSYS or HRT (Premelle) in postmenopausal women with climacteric symptoms. The Greene Climacteric Scale was used to assess the clinical effects at baseline and after 16 weeks' treatment with either JWSYS or Premelle. The physiological parameters, such as follicle-stimulating hormone and estradiol levels, were also recorded at the same time points. RESULTS: The results showed that JWSYS had a relatively lower discontinuation rate due to adverse effects, in particular the bleeding and breast tenderness. Both JWSYS and Premelle effectively alleviated most of the menopausal symptoms with no significant differences between treatment groups, whereas the beneficial effects of JWSYS were not mediated by hormone replacement-like effects. Moreover, JWSYS showed a good compliance and safety without estrogenic effects and metabolic alterations. CONCLUSIONS: It was suggested that JWSYS was a safe and efficacious therapy and might be an alternative choice for relief of climacteric symptoms in postmenopausal women. However, the exact efficacy and clinical roles of JWSYS have not been convincingly demonstrated in this study because of the blinding approach and some statistical concerns, and only the possibility of its efficacy has been raised. Therefore, a blinding trial with more patient numbers to evaluate the efficacy of JWSYS deserves further study. PMID- 12568738 TI - The European Consensus Development Conference 2002: Sex Steroids and Cardiovascular Diseases. On the route to combined evidence from OC and HRT/ERT. PMID- 12568739 TI - Relevant results of the WHI study for the management of the menopause in Spain. PMID- 12568740 TI - Metabolic flux analysis of Escherichia coli K12 grown on 13C-labeled acetate and glucose using GC-MS and powerful flux calculation method. AB - A new algorithm was developed for the estimation of the metabolic flux distribution based on GC-MS data of proteinogenic amino acids. By using a sensitive GC-MS protocol as well as by combining the global search algorithm such as the genetic algorithm with the local search algorithm such as the Levenberg Marquardt algorithm, not only the distribution of the net fluxes in the entire network, but also certain exchange fluxes which contribute significantly to the isotopomer distribution could be quantified. This mass isotopomer analysis could identify the biochemical changes involved in the regulation where acetate or glucose was used as a main carbon source. The metabolic flux analysis clearly revealed that when the specific growth rate increased, only a slight change in flux distribution was observed for acetate metabolism, indicating that subtle regulation mechanism exists in certain key junctions of this network system. Different from acetate metabolism, when glucose was used as a carbon source, as the growth rate increased, a significant increase in relative pentose phosphate pathway (PPP) flux was observed for Escherichia coli K12 at the expense of the citric acid cycle, suggesting that when growing on glucose, the flux catalyzed by isocitrate dehydrogenase could not fully fulfill the NADPH demand for cell growth, causing the oxidative PPP to be utilized to a larger extent so as to complement the NADPH demand. The GC-MS protocol as well as the new algorithm demonstrated here proved to be a powerful tool for characterizing metabolic regulation and can be utilized for strain improvement and bioprocess optimization. PMID- 12568741 TI - Function and mechanism of a low-molecular-weight peptide produced by Gloeophyllum trabeum in biodegradation of cellulose. AB - A special low-molecular-weight peptide named Gt factor, was isolated and purified via HPLC from the culture extract of the brown-rot fungus Gloeophyllum trabeum. It had high-affinity Fe(3+)-chelating ability and could reduce Fe(3+) to Fe(2+). In the presence of O(2), it could produce hydroxyl radicals HO*. The effects of Gt factor on cellulose degradation suggested that Gt factor could disrupt inter- and intra- hydrogen bonds in cellulose chains by a HO*-involved mechanism. This resulted in depolymerization of cellulose chains, which produced more reducing and non-reducing ends, thus making cellulose accessible for further degradation. This pathway was quite different from the hydrolytic processes driven by cellulases, and Gt factor might play an important role in the early stage of cellulose depolymerization by brown-rot fungi. PMID- 12568742 TI - Organic acid complexation, heavy metal distribution and the effect of ATPase inhibition in hairy roots of hyperaccumulator plant species. AB - Heavy metal uptake and distribution were investigated in hairy roots of the Cd hyperaccumulator, Thlaspi caerulescens, and the Ni hyperaccumulator, Alyssum bertolonii. Hairy roots of both species contained high constitutive levels of citric, malic and malonic acids. After treatment with 20 ppm Cd or 25 ppm Ni, about 13% of the total Cd in T. caerulescens roots and 28% of the total Ni in A. bertolonii were associated with organic acids. T. caerulescens and A. bertolonii hairy roots remained healthy and grew well at high concentrations of Cd and Ni, respectively, whereas hairy roots of the non-hyperaccumulator, Nicotiana tabacum, did not. Most of the Cd in T. caerulescens and N. tabacum roots was localised in the cell walls. In contrast, 85-95% of the Ni in A. bertolonii and N. tabacum was associated with the symplasm. Growth of T. caerulescens and A. bertolonii hairy roots was severely reduced in the presence of diethylstilbestrol (DES), an inhibitor of plasma membrane H(+)-ATPase. Treatment with DES increased the concentration of Cd in the symplasm of T. caerulescens about 6-fold with retention of root viability, whereas viability and Ni transport across the plasma membrane were both reduced in A. bertolonii. These results suggest that the mechanisms of Cd tolerance and hyperaccumulation in T. caerulescens hairy roots are capable of withstanding the effects of plasma membrane depolarisation, whereas Ni tolerance and hyperaccumulation in A. bertolonii hairy roots are not. PMID- 12568743 TI - Adhesion of plant roots to poly-L-lysine coated polypropylene substrates. AB - The ability to immobilize plant tissue in a bioreactor is an important process tool. We have shown that roots of several species rapidly attach to poly-L-lysine coated polypropylene mesh in a liquid environment. Using transformed roots of Artemisia annua as a model, the attachment process was found to be enhanced by sheep serum, but not BSA and inhibited by excess Mn(2+), but unaffected by Ca(2+) or Mg(2+). Attempts to characterize the molecule(s) responsible for binding using lectins and antibodies showed that the binding site does not appear to be glycosylated or vitronectin-like. This method of rapid attachment should prove useful for controlled immobilization of roots in bioreactors. PMID- 12568744 TI - Inhibition of paclitaxel and baccatin III accumulation by mevinolin and fosmidomycin in suspension cultures of Taxus baccata. AB - To achieve a better understanding of the metabolism and accumulation of paclitaxel and baccatin III in cell cultures of Taxus, inhibitors of the early steps in the terpenoid pathway were applied to a cell suspension culture of Taxus baccata: fosmidomycin as an inhibitor of the non-mevalonate branch of the pathway, and mevinolin as an inhibitor of the mevalonate branch. Synthesis of both taxanes in the cell suspension was first increased when cultured in the product formation medium supplemented with methyljasmonate (100 microM). The product formation medium was selected after assaying 24 different culture media. When fosmidomycin (200 microM) was added to the product formation medium together with the elicitor, the accumulation of paclitaxel and baccatin III was reduced by up to 3.0 and 1.5 times, respectively, whereas the inhibitory effect of mevinolin (1 microM) was only clearly exerted in the case of paclitaxel. Under the conditions of our experiment, we conclude that in the synthesis of both taxanes, the non-mevalonate pathway is the main source of the universal terpenoid precursor isopentenyl diphosphate (IPP). PMID- 12568745 TI - Estimation of viable biomass in aerobic biodegradation processes of organic fraction of municipal solid waste (MSW). AB - 2-(p-Iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT) dehydrogenase test and RNA assay were introduced to evaluate biomass in the processes of aerobic biodegradation of the organic fraction of municipal solid waste (MSW) in bioreactors. It was found that RNA quantification by KOH/UV method delivered reliable and repeatable results. Relative standard deviation (RSD) for INT test was significantly higher than for RNA assay and achieved values of 3 15%. Moreover, it occurred that the optimum temperature for the growth of autochthonic biomass, which takes part in the biodegradation process, was in the range from 25 to 37 degrees C. PMID- 12568746 TI - Growth at low temperature suppresses readthrough of the UGA stop codon during the expression of Bacillus subtilis flgM gene in Escherichia coli. AB - The efficient production of recombinant proteins in Escherichia coli requires a proper termination of translation to ensure the synthesis of only the desired product. During the recombinant production of Bacillus subtilis flgM in E. coli, we detected an additional polypeptide of molecular mass higher than the expected, corresponding to a product of a translational readthrough of the UGA stop codon. In this paper we show that the readthrough was abolished when the synthesis of the recombinant protein was carried out at 25 degrees C. The possible causes that contribute to reduce the proportion of readthrough protein species against the correct terminated product are discussed. PMID- 12568747 TI - Enhanced production of plumbagin in immobilized cells of Plumbago rosea by elicitation and in situ adsorption. AB - Cell cultures of Plumbago rosea were immobilized in calcium alginate and cultured in Murashige and Skoog's basal medium containing 10 mM CaCl(2) for the production of plumbagin, an important medicinal compound. Studies were carried to find out the impact of immobilization on the increased accumulation of this secondary metabolite. Immobilization in calcium alginate enhanced the production of plumbagin by three, two and one folds compared to that of control, un-crosslinked alginate and CaCl(2) treated cells respectively. Cell loading at a level of 20% to the polymer volume (Na-alginate) was optimal and maximum plumbagin was obtained. At higher cell loading (40-50%), lower plumbagin accumulation was noticed. Addition of 200 mg l(-1) chitosan as an elicitor to the immobilized cells resulted in eight and two folds higher accumulation of plumbagin over control and immobilized cells. Also, more than 70% of the plumbagin was released into the medium, which is highly desirable for easy recovery of the product. Sucrose utilization rate of the cells was higher when cells were subjected to in situ product removal using Amberlite XAD-7. This may indicate that the toxicity of plumbagin was reduced on cells when it was removed from the medium. Cells subjected to combined treatments of chitosan, immobilization and in situ extraction showed a synergistic effect and yielded 92.13 mg g(-1) DCW of plumbagin which is 21, 5.7, 2.5 times higher than control, immobilized, immobilized and elicited cells respectively. PMID- 12568748 TI - Expression and functional reconstitution of a recombinant antibody (Fab') specific for human apolipoprotein B-100. AB - We have cloned and constructed plasmid vectors, pETB23H and pETB23L, for bacterial expression of heavy (H) and light (L) chain cDNAs of Fab' of mAbB23 a monoclonal antibody specific to human plasma apolipoprotein (apo) B-100. The H- and L-chains were expressed as insoluble inclusion bodies in the cytoplasm of Escherichia coli. The inclusion bodies of both chains were isolated from the cell lysate, solubilized in 6 M guanidium-HCl, and mixed in equal molar amounts. Refolding was performed in three stages of dialysis: first, dialysis against 3 M guanidium buffer, next, continuous decrement of guanidium in the dialysis buffer through slow addition of 1 M guanidium buffer, and finally, dialysis against a buffer without guanidium. After the refolding, active Fab' (rFab') was purified through an apo B-100-coupled affinity column. When compared by ELISA, the rFab' had a slightly decreased antigen-binding activity (about 0.7-fold) compared with native Fab. The refolding yield was maximum (75%) when performed at the protein concentrations not more than 0.4 mg ml(-1), whereas the yield decreased exponentially at higher concentrations. The maximum recovery was obtained at the refolding concentration of 1.8 mg ml(-1), where the yield was about 45%. Overall, 2.4-3.0 mg of active rFab' specific to apo B-100 was successfully obtained from 1 l cultivation of E. coli cells. PMID- 12568749 TI - Incidence of Salmonella from poultry products and their susceptibility to antimicrobial agents. AB - The incidence of Salmonella in 60 samples of poultry products of national origin available for consumers obtained from two local butcher shops and one canteen of the city of Porto and the susceptibility to antimicrobial agents allowed for human or animal therapy were evaluated. The results show that poultry samples are frequently contaminated with Salmonella (60%), belonging to 10 different serotypes. Salmonella enteritidis and S. hadar were the most prevalent serotypes. In addition, a high number (75%) of the Salmonella isolates was resistant to one or more antimicrobial agents and eight different resistance profiles were recorded. Resistance to nalidixic acid and enrofloxacin was demonstrated for 50% of the isolates and the occurrence of resistant and multiresistant S. enteritidis isolates were less frequent than for S. hadar. This study suggests a high incidence of Salmonella on Portuguese poultry products and shows that they could be a potential vehicle of resistant Salmonella foodborne infections. PMID- 12568750 TI - Prevalence of Clostridium botulinum types B, E and F in faecal samples from Swedish cattle. AB - Faeces were collected from 60 cows at three slaughterhouses situated in southern and central Sweden. The faecal samples were collected during two sampling periods over the year, summer and winter. All samples were analysed for the presence of Clostridium botulinum spores, according to a combined selection and enrichment PCR procedure. One PCR assay was specific for part of the type B neurotoxin gene, while the other assay was specific for both type E and F neurotoxin genes. The prevalence of C. botulinum in Swedish cattle was established to be 73% for non proteolytic type B and less than 5% for types E and F. Twenty-eight (64%) of the positive faecal samples had a spore load of less than 4 spores/g. Statistical analysis (ANOVA) showed that seasonal variation (summer and winter) had a significant effect on the prevalence of C. botulinum type B in cattle, whereas the effect of geographical location of rearing of the cattle (southern and central Sweden) was less significant. PMID- 12568751 TI - Identification, isolation and quantification of representative bacteria from fermented cassava dough using an integrated approach of culture-dependent and culture-independent methods. AB - The use of denaturing gradient gel electrophoresis (DGGE) and traditional culture depending methods for examining the bacterial community of traditional cassava starch fermentation were investigated. It appeared that DGGE profiles of total DNA of cassava dough exhibited 10 distinguishable bands. In contrast, DGGE fingerprints of bacteria recovered from enrichment cultures of fermented dough gave variable profiles containing fewer bands. Bands corresponding to five bacterial species detected by direct PCR-DGGE of total DNA from of cassava dough were also observed in DGGE patterns of enrichment cultures. Eighteen strains were isolated from cultures selected on the basis of their DGGE banding patterns. Assessment of bacterial identification by 16S rDNA sequence similarity revealed that band comigration implied sequence identity. Comparison of 16S rDNA sequences of excised DGGE bands and recovered pure culture isolates with those in GENBANK and the RDP databases revealed that representative bacteria of fermented cassava dough were Lactobacillus and Pediococcus species as well as species of Clostridium, Propionibacterium and Bacillus. Some Lactobacillus species detected in dough samples by sequence analysis of DGGE bands were not recovered in any of the five culture media and conditions used. On the other hand, some species recovered as pure cultures from enrichments were not detected by direct DGGE analysis of total bacterial DNA from cassava dough. Our results provide evidence of the necessity to combine both culture-dependent and culture-independent methods for better description of microbial communities in indigenous cassava starch fermentations. PMID- 12568752 TI - Growths kinetics comparison of clinical and seafood Listeria monocytogenes isolates in acid and osmotic environment. AB - Comparison of pathogenic bacterial strains of clinical origin with strains of the same species isolated from the environment may be a valuable tool for microbial risk assessment, especially for foodborne pathogens. Thus, a number of Listeria monocytogenes strains responsible for human cases of listeriosis, in relation to the consumption of contaminated seafood, have been compared with "natural" L. monocytogenes strains isolated from similar seafood products. Complete factorial designs were used to assess quantitatively the growth abilities of four clinical and four seafood isolates of L. monocytogenes placed in various environmental conditions. The cells were submitted to acid and osmotic stress as they were in stationary phase (constant condition) or in exponential phase (dynamic condition). The effects and interactions of pH (5-7) and NaCl concentration (0.5 8% v/v) were studied at two growth temperatures (10 and 20 degrees C). Growth parameters (lag and generation times calculated with Gompertz equation) were used to compare the behavior of the strains with respect to the conditions of culture. The results indicated an overall weak effect of acid stress alone, whereas osmotic stress clearly affected bacterial growth and a synergic effect between these two factors was observed. Clinical strains displayed better adaptation than seafood strains in stationary phase, however, this difference was not verified in exponential phase. Low temperature (10 degrees C) usually confirmed the observations at 20 degrees C, and the differences between clinical and food strains were more pronounced. Finally, a classification of the eight strains, based on the collected data, showed three groups: (i) seafood strains, (ii) three clinical strains and (iii) the last clinical strain, alone due to its high resistance to adverse conditions. PMID- 12568753 TI - Evaluation of the effect of malt, wheat and barley extracts on the viability of potentially probiotic lactic acid bacteria under acidic conditions. AB - In this work, the effect of cereal extracts, used as delivery vehicles for potentially probiotic lactic acid bacteria (LAB), on the acid tolerance of the cells was evaluated under conditions that simulate the gastric tract. More specifically, the effect of malt, barley and wheat extracts on the viability of Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus reuteri during exposure for 4 h in a phosphate buffer acidified at pH 2.5 was investigated. In the absence of cereal extracts all strains demonstrated a significant reduction in their cell population, particularly L. plantarum. The viability of L. plantarum was improved by approximately 4 log(10) cycles in the presence of malt and 3 log(10) cycles in the presence of wheat and barley. The survival of L. acidophilus and L. reuteri was increased by more than 1.5 and 0.7 log(10) cycle, respectively, upon addition of cereal extracts. In order to evaluate the contribution of the cereal constituents on cell survival, the individual effect of glucose, maltose and free amino nitrogen (FAN), which were added at concentrations that correlated to the reducing sugar and FAN content of the cereal extracts, was examined. The viability of L. plantarum was progressively improved as the maltose or glucose concentration increased; an increase by approximately 2 log(10) cycles was observed in the presence of 8.33 g/l sugar. The survival of L. acidophilus increased by more than 1 log(10) cycle, even at very low concentrations of maltose and glucose (e.g., 0.67 g/l), while L. reuteri stability was enhanced in the presence of maltose but no appreciable effect was demonstrated in the presence of glucose. Sugar analysis indicated that glycolysis was inhibited in all cases. Addition of tryptone and yeast extract, used as sources of FAN, enhanced L. acidophilus acid tolerance, but did not affect L. reuteri and L. plantarum. The results presented in this study indicate that malt, wheat and barley extracts exhibit a significant protective effect on the viability of L. plantarum, L. acidophilus and L. reuteri under acidic conditions, which could be mainly attributed to the amount of sugar present in the cereal extracts. PMID- 12568754 TI - Quantification of the copy number of nor-1, a gene of the aflatoxin biosynthetic pathway by real-time PCR, and its correlation to the cfu of Aspergillus flavus in foods. AB - A real-time PCR system directed against the nor-1 gene of the aflatoxin biosynthetic pathway as a target sequence has been applied to detect an aflatoxinogenic A. flavus strain in plant-type foods like maize, pepper and paprika. The system is based on the TaqMan fluorescent probe technology. The copy numbers of the nor-1 gene were compared to conventional cfu data obtained from the same set of samples. In general, a good correlation between nor-1 gene copy number and the cfu data was observed; however, the nor-1 copy numbers were always higher. It was shown that the system is specific for nor-1 containing species. PMID- 12568755 TI - Evolution of microbial populations during traditional Feta cheese manufacture and ripening. AB - In three different dairies (A, B and C) located in Peloponess region (Southern Greece), traditional Feta cheese trials took place February to March using mixtures of sheep's and goat's milk. Only small variations in the evolution of microbial groups were observed during the whole ripening period. The main groups, such as thermophilic cocci, mesophilic lactococci, thermophilic lactobacilli, nonstarter lactic acid bacteria (NSLAB), presumptive Leuconostoc, enterococci and micrococci, reached their highest levels during the first 16 days, and then declined approximately 1-2 log units until the end of ripening. The remaining groups investigated, comprising yeasts, coliforms and Escherichia coli, were highest at day 4. The yeasts remained constant, while coliforms and E. coli decreased sharply and were not detectable after 120 days of ripening. A number of 146 isolates (dairy A) taken from all stages of the manufacturing and ripening process were purified and studied. Lactobacillus plantarum (58/146) and isolates of related species Lactobacillus pentosus and Lactobacillus paraplantarum (16/146) were the most common microorganisms found during cheese ripening. Streptococcus thermophilus (23/146) and Lactobacillus delbrueckii subsp. bulgaricus (20/146) were detected in high levels up to 20 days, and then gradually reduced. Enterococcus faecium (29/146) was found in all manufacturing and ripening stages. PMID- 12568756 TI - Molecular typing of Lactobacillus delbrueckii of dairy origin by PCR-RFLP of protein-coding genes. AB - Thirty-five strains of Lactobacillus delbrueckii subsp. lactis and subsp. bulgaricus isolated from dairy products were typed by restriction fragment length polymorphism (RFLP) of protein-coding genes. The strains were analysed by RFLP of PCR amplified, infragenic fragments of the following housekeeping genes: beta galactosidase, lactose permease, and proline dipeptidase. Sequencing of the variable regions of the 16S rDNA was then performed on a reduced number of strains. PCR-RFLP analysis evidenced wide strain heterogeneity. Strains were grouped into genotypes according to both subspecies assignment and infra-species genetic polymorphism. This polymorphism was related to the presence of microbial groups within subspecies populations. The low infra-species sequence polymorphism detected in the variable region of the 16S rRNA gene did not enable to group the strains with the same sensitivity reached by PCR-RFLP of protein-coding genes. PCR-RFLP of protein-coding genes applied to L. delbrueckii seems a promising tool to evaluate microbial diversity within bacterial subpopulations. Differences among bacterial subpopulations based upon molecular heterogeneity in protein coding genes would enable to better understand the role of strains from different ecological niches. PMID- 12568757 TI - The source of ochratoxin A in Brazilian coffee and its formation in relation to processing methods. AB - A total of 408 Brazilian coffee samples was examined during the 1999 and 2000 coffee harvest seasons for the presence of ochratoxin A (OA) and fungi with the potential to produce it. Samples came from four regions: Alta Paulista (western area of Sao Paulo State), Sorocabana (southwest Sao Paulo State), Alta Mogiana (northeast Sao Paulo State) and Cerrado Mineiro (western area of Minas Gerais State). Cherries and beans were examined at different stages: immature, mature and overripe cherries from trees, overripe cherries from the ground and beans during drying and storage on the farm. For mycological studies, the cherries and beans were surface disinfected with chlorine, plated on Dichloran 18% Glycerol Agar at 25 degrees C for 5-7 days and analysed for the presence of Aspergillus ochraceus and closely related species, A. carbonarius and A. niger. More than 800 isolates of fungi belonging to these species were identified and studied for the ability to produce OA using the agar plug technique and thin layer chromatography (TLC). A. niger was the species found most commonly (63% of isolates of these three species), but only 3% of them produced OA. A. ochraceus also occurred commonly (31% of isolates), and 75% of those studied were capable of OA production, a much higher percentage than reported elsewhere. A. carbonarius was found (6% of isolates) only in Alta Paulista, the hottest region studied, and only from beans in the drying yard or in storage. However, 77% of the A. carbonarius isolates were capable of producing OA. Average infection rates for cherries taken from trees were very low, but were higher in fruit taken from the ground, from the drying yard and from storage, indicating infection by toxigenic species after harvest. The average OA content in 135 samples of mature cherries from trees, overripe from trees, overripe from the ground, drying yard and storage was 0.1, <0.2, 1.6, 2.1 and 3.3 microg/kg, respectively. Although individual OA levels varied widely, only 9 of the 135 samples analysed exceeded 5 microg/kg OA, with one sample of poor quality dried coffee in excess of 100 microg/kg OA. The causes of high contamination were investigated on the farms concerned and several critical points were found, relating both to local climatic conditions and the drying processes used. PMID- 12568758 TI - Identification of lactic acid bacteria from spontaneous fermentation of 'Almagro' eggplants by SDS-PAGE whole cell protein fingerprinting. AB - In order to complete the previously performed phenotypic identification [Sanchez et al., 2000. Int. J. Food Microbiol. 59 (2000) 9], whole cell protein patterns obtained using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE) of 149 lactic acid bacteria (LAB) strains, isolated from 'Almagro' eggplants spontaneous fermentation, were analysed. Phenotypic identification of those strains had revealed the presence of the species Lactobacillus (Lb) plantarum, Lb. pentosus, Lb. brevis biotype 2, Lb. brevis biotype 3, Lb. fermentum, Lactobacillus spp. and Aerococcus viridans. The SDS-PAGE technique generated complex and stable patterns that were easy to interpret and compare with those of the 17 reference strains used in this study. After numerical analysis of the resulting electrophoretic protein patterns, six well-delineated clusters were discriminated that, with some exceptions, correlated well with the different groups phenotypically found. Only two strains showed unique protein patterns without meaningful homology to any of the reference strains used for this study and remained unidentified. PMID- 12568759 TI - Characterisation of a Bifidobacterium strain with acquired resistance to cholate- a preliminary study. AB - The susceptibility levels against bile salts (ox gall) and sodium cholate of 19 Bifidobacterium strains from different origins ranged between 0.125% and 2.0% and 0.05% and 0.4%, respectively. Seven of these strains were subsequently selected for the isolation of sodium cholate resistant derivatives by exposure to gradually increasing concentrations of this compound. Derivatives resistant to at least 1.2% of cholate were obtained from Bifidobacterium bifidum CECT 4549 and B. bifidum M6. Further analysis of the CECT 4549 derivative indicated that the phenotype induced was stable; the enzymatic activities analysed remained unchanged and no major rearrangements of chromosomal DNA were produced. However, the cells of the resistant derivative were smaller and more regular in average size than those of the original strain. In addition, the derivative had lost the ability to use lactose, fructose and galactose, and showed reduced expression levels of two membrane proteins of 78 and 114 kDa. Finally, the resistant strain displayed higher survival and lower growth inhibition in the presence of cholate than the original strain. PMID- 12568760 TI - Overview of lead remediation effectiveness. AB - A Symposium on Lead Remediation Effectiveness, sponsored by the US Environmental Protection Agency, was held at Coeur d'Alene, Idaho, USA from 22-25 May, 2000. International participants from various levels of government, educational institutions, industry, and community representatives presented papers and posters on themes that ranged from engineering practices through community participation in the remediation processes. The papers in this volume represent a global distribution of sites, especially those outside the USA. In providing an overview of the symposium and the theme of Lead Remediation Effectiveness we have drawn on information from some presentations at the symposium, besides those described in this volume. PMID- 12568761 TI - A critical evaluation of public health programs at the Bunker Hill Superfund site. AB - Since 1983, the Bunker Hill Superfund site (BHSS) has been the second largest on US Environmental Protection Agency's (EPA) National Priority List for cleanup. Contaminants include millions of tons of Pb, Cd, Hg and As. In 1974, following a bag house fire, 22.1% of young children had blood lead levels >80 microg/dl. In the early 1980s to the present, EPA initiated the cleanup of exterior residential soils and the smelter complex. In 1999, The National Geological Service confirmed that heavy metal pollution had extended from BHSS to Lake Coeur d'Alene (already known earlier) all the way to the Spokane River in Washington State via water borne tributaries linking Idaho and Washington States. This report focuses on public health programs and their results initiated by Federal and State agencies at the BHSS. These programs include blood lead screening, educational programs, exploratory dust control plans, and land transactions. These programs and their results are then evaluated, assessed and critically discussed. The conclusion of this critical evaluation assessment is that the protection of public health has not been adequately addressed or protected by Federal and State agencies. PMID- 12568763 TI - Lead remediation and changes in human lead exposure: some physiological and biokinetic dimensions. AB - This paper presents a qualitative and quantitative analysis of the various aspects of lead remediation effectiveness with particular reference to human health risk assessment. One of the key elements of lead remediation efforts at such sites as those under the Superfund program deals with populations at elevated exposure and toxicity risk in the proximity of, or at, the site of remediation, especially remediation workers, workers at other tasks on sites that were remediated down to some action level of lead concentration in soils, and groups at risk in nearby communities. A second element has to do with how one measures or models lead exposure changes with special reference to baseline and post-remediation conditions. Various biomarkers of lead exposure can be employed, but their use requires detailed knowledge of what results using each means. The most commonly used approach is measurement of blood lead (Pb-B). Recognized limitations in the use of Pb-B has led to the use of predictive Pb exposure models, which are less vulnerable to the many behavioral, physiological, and environmental parameters that can distort isolated or 'single shot' Pb-B testings. A third aspect covered in this paper presents various physiological factors that affect the methods by which one evaluates Pb remediation effectiveness. Finally, this article offers an integrated look at how lead remediation actions directed at one lead source or pathway affect the total lead exposure picture for human populations at elevated lead exposure and toxicity risk. PMID- 12568762 TI - An evaluation of recent blood lead levels in Port Pirie, South Australia. AB - The Port Pirie Lead Program commenced in 1984. The abatement program involves identification of children with elevated blood lead levels, house decontamination, soil treatment, development of heavily vegetated buffer zones around the smelter, family education and support and community education. Since 1984 the smelter has also implemented substantial new emission controls and environmental improvements. Blood lead and air monitoring programs as well as investigations of emission sources, ongoing household contamination and infant exposure mechanisms are in place. Although capillary blood lead monitoring has shown a significant decrease in the mean blood lead levels of the children, 61% of children aged 1-4 years still exceed 10 microg/dl, with 28% at or above 15 microg/dl. Re-entrainment of lead from the contaminated areas within the city is only a small contributor to air-borne lead levels compared with that from the smelter and its environs. The smelter has undertaken extensive work to reduce windborne fugitive emissions. While attempts to demonstrate reductions in air lead have been hampered by large annual variations in wind speed and direction, air lead studies have confirmed that only small losses are now arising from the stockpile area of the smelter site. Evidence suggests that worker hygiene improvements, relocation of children to lower exposure suburbs, community education, house decontamination, specific measures for individual children with elevated blood lead, and avoidance of tank rainwater have all been partially successful. A substantial investigation program has refocused intervention efforts towards reducing exposure from indoor environments during the first year of life and contributed to improved identification and ranking of ongoing smelter emission sources. PMID- 12568765 TI - The influence of soil remediation on lead in house dust. AB - Lead in house dust has long been recognized as a principal source of excess lead absorption among children at the Bunker Hill Superfund Site (BHSS) in northern Idaho. House dust lead concentration from homeowner's vacuum cleaner bags has been monitored since the epidemic of childhood lead poisoning in 1974. Geometric mean house dust lead concentrations decreased from >10000 mg/kg in 1974 to approximately 4000 mg/kg in 1975, in response to air pollution control initiatives at the defective primary lead smelter. After smelter closure, 1983 mean dust lead concentrations were near 3000 mg/kg and were most dependent on soil sources. Following emergency soil removals from public areas and roadsides and fugitive dust control efforts in the mid-1980s, house dust lead decreased by approximately 40-60% to 1200-1500 mg/kg. In 1992, a cleanup goal of 500 mg/kg dust lead community average, with no individual home exceeding 1000 mg/kg, was adopted. This goal was to be achieved by a combination of contaminated soil removals and fugitive dust control efforts throughout the 21 square mile BHSS. Continual reductions in house dust lead concentrations have been noted throughout the residential area soil cleanup. Geometric mean house dust lead concentrations averaged approximately 500-600 mg/kg from 1996 to 1999 and dropped below 500 mg/kg in 2000. Analysis of these data indicates that approximately 20% of the variance in dust lead concentrations is attributed to yard, neighborhood, and community soil lead concentrations. Since 1996, dust lead concentrations and dust and lead loading rates have also been measured by dust mats placed at entryways into the homes. Neighborhood soil lead concentrations, household hygiene, the number of adults living in the home, and the number of hours a child spends outdoors in summer explain approximately 26% of the variance in mat dust lead loading rates. It is estimated that post-remedial house dust lead concentrations will stabilize at 400-500 mg/kg, as compared to approximately 200 mg/kg in socio economically similar background communities; the difference possibly attributed to residual soil concentrations (3-6 times background), recontamination of rights of-way, tracking of non-residential mining district soils and dusts, fugitive dusts associated with the remediation, and residual structural or carpet dusts. PMID- 12568764 TI - Effect of smelter emission reductions on children's blood lead levels. AB - Trail, British Columbia has been the site of an active lead-zinc smelter for approximately 95 years. Since 1989, the community has been monitoring blood lead levels in children, studying exposure pathways and conducting comprehensive education and case management programs. From 1989 through 1996, mean blood lead levels of pre-school children declined at an average rate of 0.6 microg/dl per year. From 1996 to 1999, mean blood lead levels fell at an average rate of 1.8 microg/dl per year, from 11.5 in 1996 to 5.9 in 1999. The recent rapid decline appears to be mainly attributable to the start-up of a new lead smelter using modern flash-smelting technology in May of 1997. In 1998, the annual arithmetic mean air lead level in Trail was 0.28 microg/m(3), compared with 1.1 microg/m(3) in 1996. Reductions of approximately 50% were observed in lead loadings and concentrations in outdoor dustfall, street dust and indoor dustfall after smelter emissions were reduced. Slight reductions (statistically insignificant) have been observed in carpet dust and soil lead concentrations. During the summer of 2001, the smelting and refining operations at Trail were shut down completely for 3 months. During this period, average air lead levels in Trail dropped to 0.03 microg/m(3). The average blood lead level in Trail pre-school children at the end of the shutdown was 4.7 microg/dl. These results challenge prevailing theories about the relative importance of various environmental lead sources. For example, the US EPA Integrated Biokinetic Uptake Model for Lead (IEUBK), with its emphasis on soil concentrations, would not have predicted the dramatic decline in children's blood lead levels seen in Trail following the reductions in air lead levels. The Trail experience suggests that increased attention should be paid to the importance of active sources of highly bioavailable and mobile lead bearing dusts. PMID- 12568766 TI - Mobilization of lead from human bone tissue during pregnancy and lactation--a summary of long-term research. AB - The skeleton is potential endogenous source of lead during pregnancy and lactation. We have undertaken a longitudinal investigation into the mobilization of lead from the human maternal skeleton to determine whether lead is mobilized from the maternal skeleton during pregnancy and lactation, and if so, when and how much is released. Subjects in the study were migrants to Australia (n=15) whose skeletal lead isotopic composition (endogenous lead) was different to that prevailing in the Australian environment (exogenous lead). This migrant cohort was compared with 6 multi-generational Australian controls. Biological and environmental samples were taken pre-pregnancy where possible, throughout pregnancy and postpartum for at least 6 months. Newly-born infants of the migrant and Australian mothers were monitored for 6 months. Blood lead concentrations for the migrant mothers ranged from 1.5 to 20 microg/dl (geometric mean 2.8) and for Australian mothers ranged from 1.9 to 4.3 microg/dl (geometric mean 2.9). There was minimal change in lead isotopic composition of the Australian pregnant controls although there were increases of approximately 40% in blood lead concentration in 3 of 6 cases during the postpartum period and from 0 to 12% in the other 3. In the migrant pregnant subjects, the geometric mean skeletal lead contribution to blood lead using the isotopic composition was approximately 33% (range 10-88%) for 14 subjects using a revised estimate for exogenous lead. Skeletal contribution to blood lead during the postpartum period was significantly greater than during pregnancy (P<0.001). The skeletal contributions to blood lead are higher and the changes are more consistent in those subjects who conceived within 100 days of arrival in Australia compared with those who conceived longer than 100 days. In the migrant subjects, changes in blood lead concentration during pregnancy and postpartum varied from subject to subject with an overall 20% increase; the increases during the postpartum period were greater than during pregnancy (P<0.001). It was estimated that the amount of maternal skeletal lead mobilized during pregnancy and transferred to the infant via cord blood averaged approximately 79%. The increased skeletal contribution to blood lead is attributed to a low daily calcium intake of approximately 500 mgCa/day, a condition which was present in both migrant and Australian subjects. An ongoing clinical trial is providing a new cohort with calcium supplements. A summary of other aspects of the study is included and covers: additional flux released from the skeleton during pregnancy and postpartum; XRF bone lead results; urinary excretion of lead during pregnancy and postpartum; dietary contribution to blood lead in female adults and children; comparison of rates of exchange of lead in blood of newly-born infants and mothers; relationships of lead in breast milk to lead in blood, urine and diet of the infant and mother; changes in blood lead after cessation of breastfeeding; urinary lead isotopes during pregnancy and postpartum indicate no preferential partitioning of endogenous lead into plasma; a comparison of some aspects of the nonhuman primate and human pregnancy studies. PMID- 12568767 TI - A case study of lead contamination cleanup effectiveness at Bunker Hill. AB - A review of cleanup effectiveness at Bunker Hill Superfund Site (BHSS) has shown that yard soil cleanup is an effective tool for reducing house dust lead concentrations, thereby reducing children's blood lead levels. This review has also shown that contiguous cleanup of residences has a three-fold greater reduction of children's blood lead levels compared with cleaning only those homes where children currently reside by reducing exposures attributable to neighboring properties. This review underscores the importance of a community-wide, preventative approach to controlling lead contamination in soil and house dust. This review has further characterized the need for careful design, implementation, and perpetual maintenance of a community-wide lead cleanup. Several key areas of importance to maintain large scale mining/smelting remedies in the Bunker Hill area were analyzed and noted for further action, including: infrastructure, institutional controls for homeowner projects (post cleanup), erosion control for undeveloped hillsides with potential to impact the developed valley floor, drainage improvements and flood control, waste piles, and increasing the rate at which cleanup proceeds. Focusing on these areas is crucial to minimizing recontamination at a large scale lead cleanup. PMID- 12568768 TI - An assessment of the effectiveness of lead pollution reduction strategies in North Lake Macquarie, NSW, Australia. AB - The North Lake Macquarie area of NSW, Australia, principally the suburbs of Boolaroo, Argenton and Speers Point, has been significantly polluted by lead emissions emanating from the Pasminco Metals Smelter (Cockle Creek). A lead accessibility reduction program has been carried out in North Lake Macquarie since 1991. The primary measure of success for the program is reduced blood lead levels, particularly in children. From 1991 to 2000 average child blood lead levels have decreased from 11 to 7.5 microg/dl. However, the lead accessibility reduction programs had failed to eliminate child blood lead levels that exceed the National Health and Medical Research Council maximum goal of 10 microg/dl, and a number of children still retain blood lead levels >25 microg/dl. Many factors have contributed to this failure, notably the continued presence of airborne high lead pollutants in ambient air, dusts and soil in residential areas. Significant reductions in lead mass emissions from 92 tonnes per annum (p.a.) in 1988 to approximately 15 tonnes p.a. to September 2000 have been achieved by the smelter operator. However, the reductions have been insufficient to ensure that ambient air quality consent conditions of 1 microg/m(3) are achieved in the residential areas surrounding the smelter. Sampling by the smelter operator also provides confirmation of continued deposition of high lead dusts in residential areas. The continued dust deposition places a high burden on the local community which must maintain obsessive levels of household cleanliness in order to minimise lead uptake from dusts deposited within the home and community. Evidence exists of institutional failure that may also have limited the success of the program. These failures include the use of inappropriate standards and non-approved techniques in remediation, lack of coordination between various arms of government and an unwillingness to enforce consent conditions. The smelter operator has also failed to proactively seek changes which would minimise community harm. It is suggested that for child blood lead levels to be further reduced it may be necessary to impose added restrictions on smelter emissions. Firstly, however, current emission requirements should be mandated and methodologies for remediation of historical lead depositions should also follow accepted standards and practices to maximise the benefits of the work to the affected community. PMID- 12568769 TI - Assessing remedial effectiveness through the blood lead:soil/dust lead relationship at the Bunker Hill Superfund Site in the Silver Valley of Idaho. AB - The 21 square mile Bunker Hill Superfund Site in northern Idaho includes several thousand acres of contaminated hillsides and floodplain, a 365-acre abandoned lead/zinc smelter and is home to more than 7000 people in 5 residential communities. Childhood lead poisoning was epidemic in the 1970s with >75% of children exceeding 40 microg/dl blood lead. Health response activities have been ongoing for three decades. In 1991, a blood lead goal of 95% of children with levels less than 10 microg/dl was adopted. The cleanup strategy, based on biokinetic pathways models, was to reduce house dust lead exposure through elimination of soil-borne sources. An interim health intervention program, that included monitoring blood lead and exposures levels, was instituted to reduce exposures through parental education during the cleanup. In 1989 and 2001, 56% and 3% of children, respectively, exceeded the blood lead criteria. More than 4000 paired blood lead/environmental exposure observations were collected during this period. Several analyses of these data were accomplished. Slope factors derived for the relationship between blood lead, soil and dust concentrations are age-dependent and similar to literature reported values. Repeat measures analysis assessing year to year changes found that the remediation effort (without intervention) had approximately a 7.5 microg/dl effect in reducing a 2-year-old child's mean blood lead level over the course of the last ten years. Those receiving intervention had an additional 2-15 microg/dl decrease. Structural equations models indicate that from 40 to 50% of the blood lead absorbed from soils and dusts is through house dust with approximately 30% directly from community-wide soils and 30% from the home yard and immediate neighborhood. Both mean blood lead levels and percent of children to exceed 10 microg/dl have paralleled soil/dust lead intake rates estimated from the pathways model. Application of the IEUBK model for lead indicates that recommended USEPA default parameters overestimate mean blood lead levels, although the magnitude of over prediction is diminished in recent years. Application of the site-specific model, using the soil and dust partitions suggested in the pathways model and an effective bioavailability of 18%, accurately predicts mean blood lead levels and percent of children to exceed 10 microg/dl throughout the 11-year cleanup period. This reduced response rate application of the IEUBK is consistent with the analysis used to originally develop the cleanup criteria and indicates the blood lead goal will be achieved. PMID- 12568770 TI - Environmental conditions in the Rudnaya River watershed--a compilation of Soviet and post-Soviet era sampling around a lead smelter in the Russian Far East. AB - The Rudnaya River valley in the Russian Far East contains a rich reserve of lead, zinc and boron and has been mined for nearly 100 years. Environmental contamination related to the area's mines and lead smelter was studied for over 30 years during the Soviet era, by members of the Pacific Geographic Institute (PGI). Due to government restrictions, much of the sampling focused on contamination of the river, the air, forests, vegetation, agricultural products and soil. Source-specific samples, such as stack emissions from the smelter, and blood lead levels from the residents and smelter workers could not be obtained or were classified as State secrets. However, the data do describe the extent and severity of the environmental contamination and related public health concerns. Water discharged from the smelter averages 2900 m(3)/day (containing 100 kg of lead (Pb) and 20 kg of arsenic (As)) and leachate from area mine dumps and other industrial processes contaminates the Rudnaya River. Annual air emissions include 85 tonnes of particulates (containing 50 tonnes of Pb and 0.5 tonnes of As) and 250000 m(3) of gases high in sulfur dioxide (SO(2)), carbon monoxide (CO) and carbon dioxide (CO(2)). Vegetative stress is severe and much of this area is denuded. Pb and other metals in agricultural products suggest local produce may be dangerous for human consumption, although it is a major food source for the community. Public and occupational health indicators of basophilic stippling, respiratory disease and hair lead levels further suggest the severity of the problem. Although, descriptions of complete methodologies and procedures are often lacking, these data describe how sampling was conducted during the Soviet era and document a site with severe heavy metals contamination, especially lead, and the likelihood of related public health problems. They are relevant today as investigators employ state-of-the-art-sampling techniques and explore cleanup options under a new governmental system and challenging economic times. In the post-Soviet era, a Russian/US team sampled area soils and dusts and confirmed the severity of the environmental problems using commonly employed sampling and analysis techniques. Lead concentrations in residential gardens (476-4310 mg/kg, Gx=1626 mg/kg) and in roadside soils (2020-22900 mg/kg, Gx=4420 mg/kg) exceed USEPA guidance for remediation. Preliminary biokinetic estimates of mean blood levels (average 13-27 microg/dl) suggest pre-school children are at significant risk of lead poisoning from soil/dust ingestion. Today, the PGI, in cooperation with the industrial owners and the local health and environmental authorities, is attempting to establish long-term monitoring and pollution abatement within the constraints of their difficult economic situation. PMID- 12568771 TI - Actin strange. AB - Actin, an abundant protein forming part of the cytoskeleton, can act as a co factor for viral enzymes and could thus be the key to a potential new class of anti-viral drugs. PMID- 12568772 TI - Big gene banks: nuggets for drug discovery or fool's gold? AB - Gene-mapping studies that look for complex traits among human populations have deepened our understanding of disease causes, but do they hold promise for identifying drug targets? PMID- 12568773 TI - Cleaning up the environment. AB - Aggressive melanoma cells have been shown to overexpress proteins that modify their environment, but these proteins can be inhibited by a chemically modified tetracycline. PMID- 12568776 TI - Pharma-entomology: when bugs become drugs. PMID- 12568777 TI - Structural pharmacogenomics: the answer to antimicrobial drug resistance? PMID- 12568778 TI - Polymers in medicine; a game of chess. PMID- 12568779 TI - The European conference on computational biology. AB - Highlights from the first European Conference on Computational Biology (ECCB 2002), held in conjunction with the German Conference on Bioinformatics (GCB 2002), 6-9 October 2002, Saarbrucken, Germany. PMID- 12568780 TI - The TNF superfamily is on the TRAIL to BlyS. AB - Highlights of the 9th (Biennial) International Congress on TNF-Related Cytokines (30 October-2 November 2002; The Manchester Grand Hyatt, San Diego, CA, USA. PMID- 12568781 TI - Optimization of protein therapeutics by directed evolution. AB - Directed evolution is a broadly applicable technology platform that is ideally suited to address the need for protein optimization and to fully exploit the therapeutic potential of biologics. The approach takes advantage of the remarkable structural and functional plasticity of proteins and permits the rapid remodeling of biologics into new entities with improved functions. The ability to ameliorate virtually any characteristic of a protein can translate into significant clinical benefits, including decreased immunogenicity, higher potency, greater efficacy and improved safety profile, and can considerably increase the probability of successfully developing and commercializing biotherapeutics. PMID- 12568782 TI - Toxicokinetics in preclinical evaluation. AB - Toxicokinetic evaluation is both a regulatory and scientific requirement in the drug development process. However, as a fairly new term, its use within the industry is still confused. Toxicokinetics is the generation of kinetic data to assess systemic exposure, either as an integral component of preclinical toxicity studies, or in specially designed supportive studies. These data help to understand the relationship between observed toxicity and administered dose. They also play a role in the clinical setting, assisting in the setting of plasma limits for early human exposure and in the calculation of safety margins. PMID- 12568783 TI - Microarrays: the use of oligonucleotides and cDNA for the analysis of gene expression. AB - Completion of the human genome sequence has made it possible to study the expression of the entire human gene complement (>30,000 estimated genes). Aiding in this remarkable feat, DNA microarrays have become the main technological workhorse for gene expression studies. To date, detection platforms for most microarrays have relied on short (25 base) oligonucleotides synthesized in situ, or longer, highly variable length DNAs from PCR amplification of cDNA libraries. A third choice, long (50-80 base) oligonucleotide arrays, is now available and might eventually eliminate the use of cDNA arrays. The technology has advanced to such a point that researchers now demand microarrays that are cost-effective and have flexibility and quality assurance. Short- and long-oligonucleotide technologies offer such advantages, and could possibly become the major competing platform in the near future. PMID- 12568785 TI - Combinatorial chemistry. PMID- 12568786 TI - Flexible reproduction in wild canaries is independent of photoperiod. AB - In temperate zone species, timing of the breeding season is determined by the minimum photoperiod or "critical day length" and the development of photorefractoriness. The integration of environmental cues like food, temperature, rainfall, and behavioural interactions determines the breeding window. However, there are a number of examples for breeding activities outside this time window. Here we investigated the possible mechanisms for early seasonal breeding activities in a population of wild canaries (Serinus canaria) that inhabits an isolated island within the Madeiran archipelago. In December 1999, breeding activities were observed six weeks before the usual onset of the breeding season on the island, which can be related to unusual heavy rainfall during the two previous months. Moreover, testosterone (T) levels of the birds were significantly higher as during the same time in previous years and showed no difference to the T levels found in the breeding season. Thus, birds do not rely on the cues given by the photoperiod alone, but react independently of day-length to favourable environmental conditions, like water availability. Therefore, the individuals of this wild canary population can be considered as seasonal opportunistic breeders. PMID- 12568787 TI - Cloning and expression of aromatase in a turtle with temperature-dependent sex determination. AB - It has been hypothesized that estrogen production may play a pivotal role in the sex determination of reptiles with temperature-dependent sex determination (TSD). This hypothesis has been furthered by studies that have shown higher aromatase activity in the developing ovaries in some reptiles. However, other studies have not consistently supported this hypothesis. In the current study we addressed this issue by cloning P450 aromatase cDNA in the turtle, Trachemys scripta, and developing a quantitative competitive RT-PCR for aromatase. This assay was then used to quantify aromatase mRNA levels in adrenal-kidney-gonad complexes (AKG) during TSD. Aromatase mRNA was detected in the AKGs at both male- and female producing temperatures from the earliest stage of development sampled (stage 15), through hatching (stage 26). However, levels remained relatively constant during the thermosensitive period of TSD. Further, no significant difference was detected between male- and female-producing temperatures during the thermosensitive period. After the thermosensitive period, aromatase mRNA levels increased in females (this coincides with the period during which the ovaries are differentiating). These results are consistent with those of several previous studies of certain reptiles with TSD. The current results suggest that the expression of aromatase may not be a pivotal regulatory step in the sex determination cascade of this turtle. PMID- 12568788 TI - Developmental and diel changes in plasma thyroxine and plasma and ocular melatonin in the larval and juvenile bullfrog, Rana catesbeiana. AB - Diel variation in plasma thyroxine (T(4)), and plasma and ocular melatonin was studied in Rana catesbeiana tadpoles and postmetamorphic froglets on 12:12 and 6:18 light/dark (LD) regimens. A progressive rise in plasma T(4) initiates metamorphosis while melatonin can modulate metamorphic progress. Changes in the phase of the rhythms of these two hormones during development might influence the hormonal regulation of metamorphosis. The hormones studied exhibited LD cycle specific diel fluctuations except in froglet plasma T(4) and all hormones at prometamorphosis on 6L:18D. On 12L:12D, plasma T(4) and ocular melatonin peaked during the scotophase at prometamorphosis and early climax, whereas the plasma melatonin acrophase shifted from the light to the dark at climax. A nocturnal peak of plasma melatonin closely correlated with the onset and offset of dark appeared in the froglet, while the peak of ocular melatonin shifted to the light. Compared to 12L:12D, the peaks of the diel fluctuations on 6L:18D occurred later than on 12L:12D in synchrony with an earlier onset, and increase in length, of the scotophase. The phase of the hormone rhythms changed during metamorphosis in such a way that the peaks of melatonin had a different relationship to the T(4) peaks as development proceeded. On both LD cycles, the 24-h mean of plasma T(4) rose at climax and fell in the froglet whereas plasma melatonin decreased at climax and then rose to a high level in the froglet. After only minor changes during metamorphosis, froglet ocular melatonin levels decreased on 12L:12D and increased on 6L:18D. The findings indicate that the hormonal flux during metamorphosis has circadian aspects, which might explain variations in the response to exogenous hormone treatment at different times of the day and LD cycle-specific timing of development. A fall in plasma melatonin at climax appears to be as much a part of the hormonal changes of metamorphosis as a rise in plasma T(4). PMID- 12568789 TI - Thyroid hormone stimulates hepatic IGF-I mRNA expression in a bony fish, the tilapia Oreochromis mossambicus, in vitro and in vivo. AB - To gain more knowledge about the physiological regulation of hepatic insulin-like growth factor-I (IGF-I) production in bony fish, we examined the potential influence of thyroid hormone (T3, 3,5,3'-triiodothyronine) on the expression of IGF-I in the liver of the tilapia Oreochromis mossambicus, using in vitro and in vivo methods. The in vitro experiments were performed using a recently established primary hepatocyte cell culture and IGF-I expression was determined by means of semiquantitative RT-PCR. T3 (100 nM) significantly enhanced the synthesis rate of IGF-I mRNA in short (>8h) and long (>42h) time courses. The stimulating effect of T3 was detected already after 1h. After 4h, the IGF-I mRNA expression was more than 150% of the starting amount. In long time courses, after 6h the IGF-I mRNA value was about 170% of that in untreated cells and at the end of the experiment, it was still three times higher than in the control. In addition, the increase in IGF-I mRNA expression evoked by T3 (1 nM to 1 microM) was dose-dependent. In the in vivo approach, 10 individuals of tilapia received 4 daily intraperitoneal injections of T3 (6 microg/g body weight). IGF-I mRNA was assessed using dot blot technique with a tilapia specific IGF-I cRNA probe. The T3 treatment led to an increase of the IGF-I mRNA level up to 45% in the liver compared to the untreated animals. In conclusion, our results show that T3 directly stimulates the hepatic production of IGF-I in the tilapia in vitro and in vivo and indicate that in tilapia liver regulatory mechanisms seem to exist, as they are discussed for mammals. PMID- 12568790 TI - Corticosterone levels during post-natal development in captive American kestrels (Falco sparverius). AB - We investigated post-natal development of the adrenocortical stress-response system in captive American kestrels (Falco sparverius) by measurements of baseline and stress-induced levels of corticosterone at ages 10, 16, 22, and 28 days post-hatching. Baseline levels of corticosterone increased significantly during post-natal development and although chicks aged 10- and 16-days old exhibited comparable baseline corticosterone levels, those of 22-day-old chicks were significantly higher and those of 28-day-old chicks close to fledging were higher than all younger groups. Chicks in this study exhibited low levels of stress-induced corticosterone early in development and did not exhibit adult-type stress-induced levels of corticosterone until 22 days of age post-hatching. Finally, although baseline and stress-induced levels of 28-day-old birds were significantly higher than one-year-old adults, there was no relationship between baseline corticosterone concentrations and time to nest departure. The fact that baseline levels of corticosterone are low during early development and then increase during later development may be an adaptation to the negative effects of chronically elevated corticosterone levels and as previously noted in other studies may minimize these negative effects on rapid growth and development in young birds, potentially maximizing normal growth. The ability of even young kestrel chicks to elevate corticosterone levels in response to stress suggests that they may be able to physiologically cope with food shortages associated with unpredictable food resources which wild kestrels often face. PMID- 12568791 TI - Season-related changes in circulating androgen, brain aromatase, and perch calling in male ring doves. AB - The perch-call of ring doves (Streptopelia risoria) is related to territorial defence and mate attraction. Perch-calls are sexually dimorphic and individually different in structure. The expression of perch-call is androgen-dependent and is controlled by the action of testosterone on the preoptic-hypothalamic areas. However, it is not known whether the acoustic features of the call vary with the reproductive condition. We studied plasma androgen levels, brain steroid metabolism, and perch-calling of male ring doves kept in winter-like (low temperature, short days) or spring-like (mild temperature, long day) conditions. Circulating levels of androgen were higher in males kept on spring-like condition. Spring males spent more time in perch-calling than winter males. However, a detailed analysis of the call structure revealed no difference for any time or frequency parameter between groups. This work shows that in ring doves season-dependent variations in the circulating levels of androgen are correlated with differences in the amount of time males spend in perch-calling. In addition, the study suggests that in adult males the acoustic structure of the perch-calls is not influenced by season-related changes in androgen levels. PMID- 12568792 TI - Radioimmunoassay of prolactin for the meerkat (Suricata suricatta), a cooperatively breeding carnivore. AB - We report the development and validation of a highly specific heterologous radioimmunoassay (RIA) to measure meerkat prolactin (PRL) by using rabbit antiserum to human prolactin and canine [125I]iodo-PRL. Dilutions of meerkat pituitary standard and plasma gave parallel inhibition curves in the assay. Gel filtration of meerkat pituitary extracts and canine [125I]iodo-PRL run separately on a Sephadex G-100 generated identical peaks of activity, and Western blot analysis of meerkat pituitary extract with the human prolactin antiserum used in the RIA gave a molecular weight similar to canine prolactin (21kDa). We carried out a biological validation of the prolactin assay by administering three different doses each of sulpiride and cabergoline to adult male meerkats. Increasing doses of sulpiride and cabergoline caused substantial increases and decreases, respectively, in the plasma prolactin of the study animals as expected. Activation of the stress response in meerkats by capture and ketamine hydrochloride anesthesia caused short-term but significant increases in prolactin levels in individuals bled repeatedly. The RIA developed and described here was able to determine plasma concentrations of prolactin in all animals sampled. We conclude, however, that it will be important in all future studies to confine blood sampling times to 4-7 min after capture/administration of anesthesia to avoid the confounding effects of the stress response on prolactin levels. PMID- 12568793 TI - A thromboxane A(2) system in the Atlantic stingray, Dasyatis sabina. AB - Thromboxane B(2)(TXB(2)) is the stable metabolite of thromboxane A(2)(TXA(2)) and thromboxane B(2)-like immunoreactivity (iTXB(2)) has been identified in the plasma of the Atlantic stingray, Dasyatis sabina (0.57+/-0.03 ng/ml). Plasma levels of iTXB(2) increase if the blood is allowed to clot (3.0+/-0.27 ng/ml). When clotting occurs in the presence of indomethacin, this increase is partially inhibited (1.5+/-0.17 ng/ml), indicating the presence of a cyclooxygenase activity. Radioligand binding analysis using the TXA(2) analog [125I]BOP in isolated kidney membranes revealed a receptor of K(d)=2.88+/-0.51 nM and B(max)=25.6+/-5.9 fmol/mg protein. [125I]BOP binding was displaced by the TXA(2) receptor (TP receptor) agonists U46619 (IC(50)=106.4+/-15.7 nM) and U44069 (IC(50)=88.7+/-13.0 nM), and the antagonist SQ29548 (IC(50)=51.0+/-12.9 nM). Binding was also displaced stereoselectively by the antagonists (-)L657925 (IC(50)=18.9+/-3.8 nM) and (+)L657926 (IC(50)=2025+/-280 nM). Tissue bath studies revealed that U46619, a stable TXA(2) mimetic, elicited concentration-dependent contractions in the ventral aorta which were inhibited in a concentration dependent manner by the TP receptor antagonist SQ29548. Using a human TP receptor riboprobe, Northern blotting of mRNA isolated from the stingray kidney identified transcripts of 2.8 and 6kb. The 2.8kb transcript is similar to a 2.8kb transcript found in human cells or tissues, but the 6kb transcript may be unique. These data indicate the presence of a TXB(2)-like substance in the blood, a TP receptor in the kidney, TXA(2) biological activity in the ventral aorta, and expression of a TP receptor-like gene. PMID- 12568794 TI - Corticosterone concentrations in free-living spotted salamanders (Ambystoma maculatum). AB - A non-lethal technique for drawing repeated blood samples from spotted salamanders (Ambystoma maculatum) was used to examine sex, seasonal, and capture technique differences in the physiological stress response under natural, undisturbed conditions. Baseline and stress-induced (30 min of handling and restraint) corticosterone (CORT) concentrations were measured at night during the spring migrations into and out of the breeding pond, as well as during the fall migration to over-wintering sites. Females had significantly higher CORT concentrations than males during the spring migration toward the breeding pond, but this difference was not present when animals emerged from the pond post breeding or during the fall migration. CORT concentrations did not vary seasonally, but during the acute stress response, CORT concentrations nearly doubled during the spring inbound migration, exhibited a nearly significant increase during the spring outbound migration, and did not change in the fall. Allowing animals to sit overnight in the buckets in which they were captured elicited CORT concentrations that tended to be higher (although not statistically significant) than when animals were sampled on the night of capture. The ability to sample blood from spotted salamanders using a non-lethal technique offers an opportunity to better understand both amphibian physiology and ecology. PMID- 12568795 TI - Effects of a phytosterol mixture on male fish plasma lipoprotein fractions and testis P450scc activity. AB - Plant sterols (phytosterols) have been identified as one potential source of reproductive effects in fish living downstream of pulp mills. beta-Sitosterol, the predominant plant sterol in pulp mill effluent, has previously been shown to decrease plasma sex steroid and cholesterol levels and in vitro gonadal steroid production in fish. In this study, male brook trout (Salvelinus fontinalis) and goldfish (Carassius auratus) were exposed to a phytosterol mixture (72% beta sitosterol) via Silastic intraperitoneal implants to help elucidate the mechanisms of action of phytosterols on steroid depression. As cholesterol is exogenously supplied for gonadal steroidogenesis, changes in plasma cholesterol fractions were examined. In male brook trout, low-density lipoprotein cholesterol and triglyceride levels decreased significantly, 43 and 50%, respectively, in phytosterol-treated fish. It is improbable, however, that these decreases are linked to depressed gonadal steroidogenesis in fish. The activity of P450scc, which converts cholesterol to pregnenolone (the first step in the steroidogenic pathway), was not affected in testis mitochondria isolated from brook trout or goldfish. Further investigation of the mechanisms of action of phytosterols is required. PMID- 12568796 TI - C-type natriuretic peptide of rainbow trout (Oncorhynchus mykiss): primary structure and vasorelaxant activities. AB - Natriuretic peptides (NPs) play important roles in osmoregulatory and cardiovascular systems of vertebrates. For functional studies of NPs, rainbow trout (Oncorhynchus mykiss), a euryhaline fish, is an interesting model. The information on homologous NPs of salmonid fish is, however, still incomplete with respect to C-type NP (CNP). In this study, we isolated cDNAs encoding the precursor of CNP from the brain of trout. Predicted mature CNP (CNP-22) sequence was identical to that of killifish Fundulus heteroclitus, and only one amino acid was different from that of the eel Anguilla japonica, demonstrating a greater conservation among different teleost species than is found with atrial NP (ANP) and ventricular NP (VNP). While the preprosegment of trout CNP retained 57% similarity to the eel sequence, similarities were low to those of sharks and tetrapods. The major site of expression identified by RT-PCR was the brain with minor expression in the atrium. The putative mature CNP-22 was synthesized and its biological activity was compared with other trout NPs (ANP and VNP) using trout ventral aorta, efferent branchial and celiacomesenteric arteries and anterior cardinal vein in vitro. Synthetic trout CNP-22 relaxed all pre contracted vessels with potencies comparable to trout ANP and VNP. PMID- 12568797 TI - Characterization of a cDNA encoding P-450 aromatase (CYP19) from Japanese eel ovary and its expression in ovarian follicles during induced ovarian development. AB - A cDNA encoding P450 aromatase (CYP19) was isolated from a Japanese eel (Anguilla japonica) ovarian cDNA library. This cDNA contains a complete open reading frame encoding 511 amino acid residues. The deduced amino acid sequence is 59% and 65% identical to the catfish and rainbow trout forms, respectively, and 52-54% to mammalian and chicken forms. Non-steroidogenic COS-7 cells transfected with the eel CYP19 cDNA converted exogenous androstenedione to estrone, thus verifying its identity. Northern blot analysis indicated that there was a single 2.1 kb transcript in the ovary. A 2.1 kb transcript was also found in the brain but not in the spleen, head kidney, kidney, or liver. Throughout ovarian development induced by weekly injections of salmon pituitary homogenate (SPH, 20 microg/g body weight), the 2.1 kb transcript was barely or not detectable in the ovaries. However, signals greatly increased in intensity in oocytes in the migratory nucleus stage and then decreased slightly in the post-ovulatory ovary. These changes in transcript levels are consistent with the changes in aromatase activity of ovarian follicles, suggesting that aromatase activity in ovarian follicles is mainly regulated at the transcriptional level. In addition, fadrozole was found to significantly inhibit aromatase activity in a heterologous expression system using COS-7 cells, which indicates that fadrozole treatment could be useful to control E(2) production during artificial maturation of eels. PMID- 12568798 TI - Genomic structure of the luciferase gene and phylogenetic analysis in the Hotaria group fireflies. AB - The luminescent fireflies have species specific flash patterns, being recognized as sexual communication. The luciferase gene is the sole enzyme responsible for bioluminescence. We describe here the complete nucleotide sequence and the exon intron structure of the luciferase gene of the Hotaria-group fireflies, H. unmunsana, H. papariensis and H. tsushimana. The luciferase gene of the Hotaria group firefly including the known H. parvula spans 1950 bp and consisted of six introns and seven exons coding for 548 amino acid residues, suggesting highly conserved structure among the Hotaria-group fireflies. Although only one luciferase gene was cloned from H. papariensis, each of the two sequences of the gene was found in H. unmunsana (U1 and Uc) and H. tsushimana (T1 and T2). The amino acid sequence divergence among H. unmunsana, H. papariensis, and H. tsushimana only ranged from zero to three amino acid residues, but H. parvula differed by 10-11 amino acid residues from the other Hotaria-group fireflies, suggesting a divergent relationship of this species. Phylogenetic analysis using the deduced amino acid sequences of the luciferase gene resulted in a monophyletic group in the Hotaria excluding H. parvula, suggesting a close relationship among H. unmunsana, H. papariensis and H. tsushimana. Additionally, we also analyzed the mitochondrial cytochrome oxidase I (COI) gene of the Hotaria group fireflies. The deduced amino acid sequence of the COI gene of H. unmunsana was identical to that of H. papariensis and H. tsushimana, but different by three positions from H. parvula. In terms of nucleotide sequences of the COI gene, intraspecific sequence divergence was sometimes larger than interspecies level, and phylogenetic analysis placed the three species into monophyletic groups unresolved among them, but excluded H. parvula. In conclusion, our results suggest that H. unmunsana, H. papariensis and H. tsushimana are very closely related or might be an identical species, at least based on the luciferase and COI genes. PMID- 12568799 TI - Antioxidant capacity in rat skeletal muscle tissues determined by electron spin resonance. AB - The amount of radical scavenging activity in muscle is unknown. The present study examines whether electron spin resonance (ESR) could measure and distinguish antioxidant capacity in muscle with different contractile and metabolic characteristics. Specimens of the soleus, plantaris, gastrocnemius (deep/surface portions), heart and diaphragm were obtained from female Wistar rats (n=7; 12 weeks old). Scavenging activity against superoxide anions in these specimens were determined by ESR using a spin-trapping chemical (5,5-dimethyl-1-pyrroline-N oxide). The ESR signal intensity of reaction mixtures containing muscle tissues was significantly lower in the heart, soleus, diaphragm and deep portion of the gastrocnemius than in the plataris and surface portion of the gastrocnemius. Thus, the amount of scavenging activity converted into superoxide dismutase activity was the highest in the heart, and higher in the soleus, diaphragm and deep portion of the gastrocnemius than in other muscles (ANOVA, P<0.01). In addition, scavenging activity significantly correlated with citrate synthase activity (r=0.72, P<0.01, n=42) and myoglobin content (r=0.63, P<0.01, n=42). These findings suggested that ESR and spin-trapping can be detect differences in free radical scavenging activity among muscle tissues with different metabolic characteristics. PMID- 12568800 TI - Storage of retinal in the eggs of the ascidian, Halocynthia roretzi. AB - Retinoids in the eggs of the solitary ascidian, Halocynthia roretzi, were analyzed by high performance liquid chromatography. Retinal was the almost exclusive retinoid (>>99%), and the concentration of retinal was 25.9-40.1 (30.6 on average) ng/mg of protein. The egg retinal consisted of four isomers: all trans (50.9%), 9-cis (6.8%), 11-cis (20.4%) and 13-cis (21.9%). The presence of retinal in the eggs of this ascidian is a characteristic shared with the wide range of oviparous vertebrates, although the isomer composition differs between ascidian eggs and vertebrate eggs; in vertebrate eggs, almost all the retinal is in the all-trans form. The egg retinal was bound to a protein complex via a Schiff base linkage. The electrophoretic characteristics of the protein complex were similar to that of egg yolk proteins of oviparous vertebrates. The results presented in this study strongly suggest that, as is found with oviparous vertebrates, retinal in the ascidian eggs is the essential mode of retinoid storage, and is the precursor of photoreceptive pigment chromophores and retinoic acid during development. PMID- 12568801 TI - Partial purification and characterization of midgut leucyl aminopeptidase of Morimus funereus (Coleoptera: Cerambycidae) larvae. AB - Exopeptidases of Morimus funereus larvae were partially purified and characterized. Specific leucyl aminopeptidase (LAP) activity was increased eight fold by gel filtration of the crude midgut extract. The partially purified LAP had a molecular mass greater than 100 kDa with pH optima from 7.0-9.0 and no strict substrate specificity. M. funereus LAP preferentially hydrolyzed p nitroanilides with hydrophobic amino acids in the active site, with a K(m) for leucine-p-nitroanilide of 0.21 mM. Zymogram analysis of an electropherogram obtained by native polyacrylamide gel electrophoresis revealed four enzymatically active proteinases using leucine-p-nitroanilide and methionine-p-nitroanilide as substrates and two enzymatically active proteinases using lysine-p-nitroanilide as a substrate. Although the optimal temperature of LAP activity was 40 degrees C, the enzyme was active over a broad temperature range from 2 to 60 degrees C. Among a number of inhibitors tested, heavy metals and 1,10-phenanthroline completely inhibited the enzyme, while methanol, ethanol and EGTA stimulated somewhat LAP activity. PMID- 12568802 TI - The process of hypoxic induction of Daphnia magna hemoglobin: subunit composition and functional properties. AB - The process of oxygen-dependent hemoglobin induction in Daphnia magna was studied over an 11-day period of hypoxia (ambient oxygen partial pressure: 3 kPa). Along with the increase of hemoglobin concentration in the hemolymph, hemoglobin became the dominant protein fraction in gel filtration experiments using extracts of whole animals. The size of the native aggregates was constant. However, subunit composition depended on the duration of hypoxia: the pattern of predominantly expressed subunits under hypoxia deviated from that of normoxic individuals. The varying degree of hypoxic induction for different hemoglobin subunits was confirmed by autoradiography. Along with changes in hemoglobin subunit composition, oxygen affinity of the respiratory protein increased. The dynamics of the hemoglobin induction process was analysed. Newly synthesized hemoglobin can be detected within 18 h after the onset of hypoxia. A marked increase in hemoglobin concentration is evident from the third day of hypoxia, and a steady state of hemoglobin concentration is reached within 11 days. The changes of hemoglobin subunit expression in response to hypoxia form the structural basis for the observed adjustments of hemoglobin function leading to enhanced oxygen transport at low ambient oxygen concentrations. PMID- 12568803 TI - Vertical stratification of fatty acids in the blubber of southern elephant seals (Mirounga leonina): implications for diet analysis. AB - Fatty acid signature analysis (FASA) is a powerful ecological tool that uses essential fatty acids (FA) from the tissues of animals to indicate aspects of diet. However, the presence of vertical stratification in FA distribution throughout blubber complicates the application of FASA to marine mammals. Blubber biopsy samples were collected from adult female southern elephant seals (Mirounga leonina) from Macquarie Island (n=11), and blubber cores were divided into inner and outer sections to determine the degree to which the blubber layer was stratified in FA composition, we found 19 FA from both blubber layers in greater than trace amounts (>0.5%). The inner and outer blubber layers could be separated using principal components analysis based on the relative proportion of FA in each layer. Dietary polyunsaturated fatty acids (PUFA) were also observed in significantly higher proportions in the inner blubber layer. Due to the degree of FA stratification in southern elephant seals, we concur with other marine mammal studies that sampling only the outer blubber layer will result in a loss of recently accumulated information regarding diet structure (as indicated by 'surplus' PUFA from the diet). This finding suggests that differential mobilization/deposition of certain FA may result in a modified signature from prey to predator. Thus, sampling animals to recover the inner blubber layer is important for studies attempting to describe aspects of marine mammal diet. This can be achieved in animals such as pinnipeds where the whole blubber layer can be readily sampled. PMID- 12568804 TI - Dietary vitamin E supplementation affects tissue lipid peroxidation of hybrid tilapia, Oreochromis niloticus x O. aureus. AB - A feeding trial was conducted to evaluate the effects of dietary vitamin E contents on the growth, ascorbate induced iron-catalyzed lipid peroxidation in post-mortem muscle and liver tissue, and Raman spectral changes in lens of juvenile hybrid tilapia (Oreochromis niloticus x O. aureus). Experimental fish were fed practical diets supplemented with 0, 50, 100, 200, 450 and 700 mg alpha tocopheryl acetate/kg diet for 14 weeks. There was no significant difference in weight gain, feed conversion ratio and protein efficiency ratio among fish fed test diets (P>0.05). Protein content of fish fed diet containing the lowest vitamin E level was the lowest (P<0.05) among all groups. No difference was found in other body constituents among test fish (P>0.05). The thiobarbituric acid reactive substances produced by iron-catalyzed lipid peroxidation in muscle and liver tissue of fish fed the diet without alpha-tocopheryl acetate supplementation were significantly (P<0.05) greater than those from fish fed diets containing higher levels of alpha-tocopheryl acetate. Dietary vitamin E supplementation increased the antioxidant capability of tilapia tissues against lipid peroxidation. Further, dietary vitamin E supplementation also influenced the lens cortical membrane structure of tilapia. PMID- 12568805 TI - Effects of cytidine 5'-diphosphocholine on plasma homocysteine levels in rat. AB - We have investigated the effects of cytidine 5'-diphosphocholine (CDP-choline) on total plasma homocysteine concentration in male Sprague-Dawley rats of 2 months of age (young rats) or 15 months of age (old rats). Oral administration of 0.35 or 1 g/kg of CDP-choline to young rats significantly increased homocysteine, by 19 and 47%, respectively (P<0.05) in plasma obtained 25 min after treatment. This effect was transient for the administration of 0.35 g/kg and increased up to 64% (P<0.05) after 150 min for the administration of 1 g/kg. However, treatment through a supplemented diet resulting in an average daily intake of 0.35 g/kg of CDP-choline for up to 60 days did not significantly alter homocysteine concentration. Old rats showed a significantly (P<0.05) lower homocysteine level (25%) than control young animals, even after 60 days of treatment with the supplemented diet. Thus, when rats are used in experimental studies on the beneficial effects of CDP-choline, it has to be considered that administration of high doses of CDP-choline will not affect the plasma levels of the risk factor homocysteine as long as the compound is not administered as a single bolus. PMID- 12568806 TI - Dependence of the coupling of dopamine receptors to G proteins on the protein redox state in the neural plasma membranes of pond snail. AB - Binding analysis using [3H]dopamine has shown that reduction of protein thiol groups with dithiothreitol (DTT) led to a dual effect on the receptors. First, the amount of dopamine-binding sites on the membranes and their affinity to the ligand were decreased. Second, the affinity of the receptors to [3H]dopamine was enhanced in the presence of GDP. Binding of D(1) antagonist [3H]SCH23390 to dopamine receptors increased following DTT treatment, opposite to the case with D(1) agonist [3H]SKF38393. The displacement of [3H]GDP by GTPgammaS was depressed by dopamine. Stimulation of [3H]GDP binding by dopamine was potentiated after incubation with DTT. Membrane nitrosylation eliminated the reciprocal dependence of GDP and dopamine binding to the membranes. It is suggested that binding of dopamine to the receptors can lead to both stimulation and inhibition of G protein activity, and the ratio of these effects depends on the reduction and oxidation of sulfhydryl groups of membrane proteins. Thiol reduction potentiated inhibitory action of dopamine receptors on coupled G proteins, and nitrosylation led to their uncoupling. PMID- 12568807 TI - Variation in lipid classes and fatty acid composition of salmon shark (Lamna ditropis) liver with season and gender. AB - The influence of season and gender on lipid content, lipid classes, and fatty acid compositions was assessed in livers of salmon shark (Lamna ditropis), caught in the Pacific Ocean. No significant difference in the hepatosomatic index was noted with season, though the lipid content was significantly higher (P<0.05) in winter. Triacylglycerol (TAG) was identified as the predominant lipid class (78.5 82.0%), followed by sterol esters (5.7-9.1%) and hydrocarbons (3.4-5.4%). No significant differences were observed in TAG composition with respect to the season or gender. However, diacylglyceryl ether contents were significantly higher (P<0.05) in winter (3.8-5.3%) than those obtained in summer (1.3-1.1%). Polyunsaturated fatty acids constituted the major fatty acid class of salmon shark total liver lipid and docosahexaenoic acid (C22:6n-3) (22.7-28.4%) was the most abundant fatty acid which was significantly lower (P<0.05) in winter. These results suggested that lipid characteristics of salmon shark liver were influenced by season, but not by gender. PMID- 12568808 TI - Differential expression of ubiquitin and proteasome-dependent pathway components in rat tissues. AB - The ATP-ubiquitin-dependent pathway in eukaryotes is a complex system, which plays an essential role in selective protein degradation. The functional diversity of this system must be matched to the specific protein metabolism related to the physiology of each cell types. The aim of our work was to study the expression of different components of the proteasome-dependent pathway in various rat tissues. Therefore we quantified the 20S proteasome and the 19S and 11S regulators by Western blot, and measured the expression of the mRNAs of certain subunits, which are markers of these components. We compared the peptidase activities of the purified 20S proteasomes, and also mapped its components by 2D electrophoresis. Our results show that the components of the ATP ubiquitin-dependent pathway vary considerably both in abundance and activity from one tissue to another. This diversity allows the cells to respond appropriately to tissue-specific protein metabolism in the rat. PMID- 12568809 TI - Purification and characterization of aspartate racemase from the bivalve mollusk Scapharca broughtonii. AB - High concentrations of D-aspartate occur in blood shell Scapharca broughtonii (Mollusca) tissues. We purified aspartate racemase from the foot muscle of the bivalve to electrophoretic homogeneity. The molecular mass shown by sodium dodecyl sulfate polyacrylamide gel was 39 kDa, while that shown by gel filtration ranged from 51 to 63 kDa. Pyridoxal 5'-phosphate-dependency of the enzyme was demonstrated by its absorption spectrum as well as the effects of amino oxyacetate and other reagents on the activity and spectrum. The enzyme is highly specific to aspartate and does not racemize L-alanine, L-serine and L-glutamate. It showed the highest activity at pH 8 both in the conversion of L- to D- and D- to L-aspartate, and the optimal temperature was 25 degrees C. V(max) and K(m) values for L-aspartate were 7.39 micromolmin(-1)mg(-1) and 60.4 mM and those for D-aspartate were 22.6 micromolmin(-1)mg(-1) and 159 mM, respectively. PMID- 12568810 TI - Differential expression of Dlk-1 in bovine adipose tissue depots. AB - Dlk-1, a type 1 membrane glycoprotein, is a member of the Epidermal Growth Factor like family of homeotic proteins that are typically involved in cell fate decisions and in mice it has been implicated in the control of differentiation of adipocytes. The aim of this study was to determine whether there were tissue specific expression patterns of Dlk-1 splice variants in bovine tissues. Only the Dlk-1-C2 variant was expressed in adult bovine tissues while both Dlk-1-C2 and Dlk-1-A variants were expressed in foetal tissues. Quantitative real-time PCR revealed large differences in the relative levels of expression of the Dlk-1-C2 variant in adult adipose tissue depots with no expression in subcutaneous and brisket adipose tissues. Expression was also demonstrated in three adult skeletal muscle samples. The large variation in the level of expression of Dlk-1-C2 in different adult tissues may reflect the relative preadipocyte content of those tissues and consequently their potential for generating new adipocytes. A low abundance soluble glycoprotein (bFA1) was purified from bovine amniotic fluid. Analyses of its amino acid sequence revealed that it corresponded to most of the extracellular domain of bovine Dlk-1 and was derived by proteolytic processing from the full-length Dlk-1 protein encoded by the Dlk-1-A variant. The tissues expressing the Dlk-1-A variant have not been identified but are likely to be foetal in origin. Splice variants of Dlk-1 may have varied functional roles with the foetal Dlk-1-A form capable of generating a protein that undergoes proteolytic processing to release a soluble ecto-domain of Dlk-1. In contrast the Dlk-1-C2 splice variant codes for a protein lacking this processing site and therefore it probably remains bound to the cell membrane. PMID- 12568811 TI - Polyunsaturated fatty acid profiles of whole body phospholipids and triacylglycerols in anadromous and landlocked Atlantic salmon (Salmo salar L.) fry. AB - We compared the fatty acid compositions and gains of whole body triacylglycerols (TAG) and phospholipids (PL) in anadromous and landlocked Atlantic salmon (Salmo salar) fry, of the same age, fed the same commercial marine oil-rich diet over a 42-day feeding trial. The landlocked strain exhibited significantly (P<0.05) higher growth rate and feed efficiency, due principally to a higher fat retention, particularly of monounsaturated and saturated fatty acids (SFA). n-3 and n-6 long-chain polyunsaturated fatty acid (PUFA) gains and retentions were significantly higher (P<0.05) in the landlocked fry. Great similarities were found in the fatty acid profiles of whole body TAG of both strains. However, marked genotypic differences were observed in the PUFA profiles of whole body PL fractions. The total PUFA, n-3 PUFA and docosahexaenoic acid (DHA) level in PL was significantly higher (P<0.05) while the SFA level, and the PUFA C18/C20 and eicosapentaenoic acid/arachidonic acid ratios were significantly lower (P<0.05) in the anadromous fry than in landlocked fry. Our results indicate that the level of DHA in salmon PL is under strong genetic control and that the capacity for incorporation, and possibly for the conversion of dietary n-3 and n-6 PUFA, is higher in the landlocked strain. PMID- 12568812 TI - The effects of dietary lipid and strain difference on polyunsaturated fatty acid composition and conversion in anadromous and landlocked salmon (Salmo salar L.) parr. AB - Five experimental diets containing different proportions of olive, sunflower and linseed oils were used in a 55-day feeding trial on both anadromous and landlocked parr of Atlantic salmon (Salmo salar) of the same age, in order to study the effects of diet and strain on growth and fatty acid composition and absolute gains in fish whole body triacylglycerols (TAG) and phospholipids (PL). Growth rate was higher in landlocked than in anadromous parr, but not between the different diets. By contrast, the effect of diet on whole body fatty acid composition was much more pronounced than that of strain difference. The fatty acids deposition results establish significant (P<0.05) positive correlations and linear relationships between the percentage of several fatty acids (18:1n-9, 18:2n-6, 18:3n-3) in dietary lipids and their absolute gains in whole body TAG and PL of both stocks. They also indicate the selective deposition of 18:1n-9 compared with linoleic acid (LLA) and linolenic acid (LNA). Finally, the results suggest the occurrence of the conversion of LLA and LNA to long-chain polyunsaturated fatty acids, its stimulation by increased substrate availability, a significantly higher n-3 and n-6 polyunsaturated fatty acids conversion capacity in landlocked than in anadromous parr and a strong genetic influence on docosahexaenoic acid content in salmon parr PL. PMID- 12568813 TI - Seasonal changes in lipid classes and fatty acid composition in the digestive gland of Pecten maximus. AB - Seasonal variations in lipid classes and fatty acid composition of triacylglycerols and phospholipids in the digestive gland of Pecten maximus were studied over a period of 16 months. Acylglycerols predominated (19-77% of total lipids), in accordance with the role of the digestive gland as an organ for lipid storage in scallops. Seasonal variations were mainly seen in the acylglycerol content, while phospholipids (2.5-10.0% of total lipids) and sterols (1.9-7.4% of total lipids) showed only minor changes. The most abundant fatty acids were 14:0, 16:0, 18:0, 16:1(n-7), 18:1(n-9), 18:1(n-7), 18:4(n-3), 20:5(n-3) and 22:6(n-3) and these showed similar seasonal profiles in both, triacylglycerol and phospholipid fractions. In contrast to the phospholipid fraction, the triacylglycerol fraction contained more 20:5(n-3) than 22:6(n-3). In three phospholipid samples we noted a high percentage of a 22-2-non-methylene interrupted fatty acid, previously described to have a structural role in several bivalve species. The main polyunsaturated fatty acids displayed important seasonal variations parallel to those of the acylglycerols, suggesting good nutritional conditions. A positive correlation existed between the level of saturated fatty acids and temperature, whereas the levels of polyunsaturated fatty acids correlated negatively with temperature. PMID- 12568814 TI - Rabbit liver dCMP phosphohydrolase: a pyrimidine 5'-nucleotidase I-like enzyme in non-erythrocytic cells. AB - A nucleotide phosphomonoesterase activity that preferably hydrolyzed dCMP was detected in rabbit liver and purified approximately 20-fold. The enzyme was similar in the catalytic and molecular properties to pyrimidine 5'-nucleotidase subclass I (P5N-I), which distributed specifically in vertebrate erythrocytes. In addition to liver, the activity was found in rabbit kidney, spleen, heart, intestine, but was not detected in any rat or chicken tissues tested. The rabbit enzyme protein reacted with antibodies against chicken P5N-I. Its pI was estimated to be approximately 5.3, and the enzyme was concluded to consist of single polypeptide of an approximately 38 kDa based on gel filtration and Western blot analysis. The partially purified enzyme preferentially hydrolyzes dCMP, UMP and CMP, K(m) values for these substrates are approximately 0.3 mM, the optimal pH is approximately 7, and the enzyme requires Mg(2+). This nucleotidase may contribute to the regulation of intracellular pyrimidine nucleotides in the rabbit. PMID- 12568815 TI - Molecular characterization of hemoglobin alpha-D chains from Geochelone carbonaria and Geochelone denticulata land turtles. AB - In order to help elucidate the evolution of alpha-globins, the complete cDNA and amino acid sequences of Geochelone carbonaria and Geochelone denticulata land turtles alpha-D chains have been described. In G. carbonaria, the cDNA is 539 bp with ATG start codon located at position 46, TGA stop codon at position 469 and AATAAA polyadenylation signal at position 520. In G. denticulata, the cDNA is 536 bp with ATG start codon located at position 46, TGA stop codon at position 469 and AATAAA polyadenylation signal at position 517. Both cDNAs codify 141 amino acid residues, differing from each other in only four amino acid residues. When comparing with human Hb alpha-chain, alterations in important regions can be noted: alpha110 Ala-Gly, alpha114 Pro-Gly, alpha117 Phe-Tyr and alpha122 His-Gln. There is a high homology between the amino acids of these turtles when compared with chicken alpha-D chains, progressively decreasing when compared with human, crocodile, snake, frog and fish alpha-chains. Phylogenetic analysis of alpha-D chains shows that those of turtles are closer to those of birds than to snakes and lizards. PMID- 12568816 TI - Caregiving and risk of coronary heart disease in U.S. women: a prospective study. AB - BACKGROUND: A growing number of women provide care to disabled or ill relatives. Many studies have linked caregiving to psychiatric morbidity, lower perceived health status, elevated blood pressure, and poorer immune function. However, no studies have examined the association between caregiving and cardiovascular disease incidence. METHODS: We conducted the study in 54,412 women from the Nurses' Health Study, a prospective cohort of female registered nurses residing in 11 U.S. states. These women were aged 46 to 71 years and did not have diagnosed coronary heart disease (CHD), stroke, or cancer at baseline (1992). We collected information on caregiving responsibilities in 1992 and coronary heart disease between baseline (June 1, 1992) and return of the 1996 questionnaire. RESULTS: During 4 years of follow-up, we documented 321 incident cases of CHD (231 nonfatal cases of myocardial infarction and 90 CHD deaths). In multivariate analyses controlling for age, smoking, exercise, alcohol intake, body mass index, history of hypertension, diabetes mellitus, and other covariates, caregiving for disabled or ill spouse for > or =9 hours per week was associated with increased risk of CHD (RR, 1.82; 95% confidence interval, 1.08-3.05). However, caregiving for disabled or ill parents or disabled or ill others was not significantly associated with increased risks of CHD. CONCLUSION: These data indicate that high levels of caregiving burden for ill spouses may increase the risk of CHD among women. PMID- 12568817 TI - Public perceptions about prematurity: a national survey. AB - BACKGROUND: More than 460,000 preterm babies are born annually in the United States, with rates varying depending on the mother's race/ethnicity. Preliminary evidence suggests that the general public may have a lack of knowledge and misconceptions about preterm birth. METHODS: A national telephone survey of U.S. adults, over-sampling for black and Hispanic women, was conducted in 2002 to assess people's knowledge, attitudes, and beliefs related to preterm birth (N=1967). Analyses included racial/ethnic differences among women's responses and comparisons to men's responses. RESULTS: Prematurity was named by <1% of women as the most important issue facing pregnant women and infants. Overall, 34% of women and 31% of men felt that prematurity is a common problem in the United States, and 38% of women and 25% of men believed prematurity to be a serious problem in the United States. More black and Hispanic women considered prematurity to be common and serious. A majority of women (65%) and men (59%) attributed preterm births to risky prenatal maternal behaviors. CONCLUSIONS: Although nearly one in eight babies is born prematurely, most U.S. adults do not consider prematurity to be a serious public health problem. While the etiology can be identified in only half of spontaneous preterm births, the public largely blames the mother's prenatal behavior. Misperceptions are prevalent and may impede future research and prevention efforts if not corrected. PMID- 12568818 TI - Research priorities for evaluating family history in the prevention of common chronic diseases. PMID- 12568819 TI - Family history assessment: strategies for prevention of cardiovascular disease. AB - Family history assessment can be used to combine population-wide health promotion and risk-reduction efforts with a high-risk, targeted approach to help reduce the burden of cardiovascular disease (CVD). Family history is an independent predictor of CVD, and the upper portion of the family history distribution explains a larger fraction of CVD in the population than can be explained by extreme values of other risk factors (e.g., blood pressure and cholesterol). A positive family history of disease captures the underlying complexities of gene gene and gene-environment interactions by identifying families with combinations of risk factors, both measured and unmeasured, that lead to disease expression. Family history is a useful tool for identifying most prevalent cases of CVD and for population-wide disease-prevention efforts. A positive family history also identifies the relatively small subset of families in the population at highest risk for CVD who may benefit most from targeted screening and intensive intervention. PMID- 12568820 TI - Family-centered approaches to understanding and preventing coronary heart disease. AB - Family history represents the contributions and interactions of unique genomic and ecologic factors that affect the metabolic profile and life course of a family and its members. It is well known that a family history of coronary heart disease (CHD) is a significant predictor of an individual's risk for CHD even after adjusting for an individual's own established risk factors, such as hypertension, smoking, and abnormal lipoprotein levels. The explanation for the observed familial disease aggregation is not well understood except for the general knowledge that genetic and environmental factors predisposing to CHD also aggregate in families. Given the multifactorial nature of an individual's risk, it can be argued that an individual's familial risk of disease may, in fact, be a better indicator of the many complex interactions among predisposing genetic and environmental factors than can be captured by an individual's own risk factors. Issues of how to assess, quantitate, and apply family history information in clinical settings still need to be resolved. Some clinical risk indicators, such as the National Cholesterol Education Program III guidelines, take into account family history, while others, such as the Framingham Risk Score, do not. Moreover, several family-centered intervention studies have demonstrated the particular advantages of focusing on families rather than just individuals. Although there has been tremendous progress in primary prevention of CHD over the last 20 years, substantial advancements may still be achieved by focusing on the family as its own unit of inference and as a specific target for disease prevention. PMID- 12568821 TI - Family history of diabetes as a potential public health tool. AB - Given the substantial morbidity and mortality associated with type 2 diabetes, it is important that public health seek ways to delay or prevent the onset of this condition. Risk factors for type 2 diabetes are well established and include underlying genetic susceptibility. Despite this knowledge, as well as significant advances in understanding the human genome, the prevalence of type 2 diabetes continues to rise at an alarming rate. Because type 2 diabetes is a complex condition involving a combination of genetic and environmental factors, DNA testing for susceptibility genes is not yet warranted. However, because family history reflects genetic susceptibility in addition to other factors, it may be a useful public health tool for disease prevention. When evaluating family history as a public health tool, several important issues need to be considered, including the analytic and clinical validity and the clinical utility of using family history as a screening tool. These issues as well as a review of the epidemiologic evidence evaluating family history as a risk factor will be reviewed.Overall, a family history approach appears to be a promising new public health tool to fight the growing epidemic of diabetes in the United States. Adequate levels of funding to further evaluate this approach and to develop appropriate tools should be made available for research activities focused on this important area. PMID- 12568822 TI - Family history as a predictor of asthma risk. AB - Asthma, one of the most important chronic diseases of children, disproportionately affects minority and low-income children. Many environmental risk factors for asthma have been identified, including animal, mite, and other allergens; cigarette smoke; and air pollutants. Genetics also play an important causative role, as indicated by familial aggregation and the identification of candidate genes and chromosomal regions linked to asthma risk. Using a positive family history of asthma to identify children at increased risk could provide a basis for targeted prevention efforts, aimed at reducing exposure to environmental risk factors. To assess the predictive value of family history as an indicator of risk for childhood asthma, we reviewed population-based studies that evaluated family history of asthma and atopic disease in children with asthma. Our search identified 33 studies from all geographic regions of the world for review. The studies varied in definitions of positive family history and asthma phenotype and used study populations with asthma prevalence ranging from 2% to 26%. Nevertheless, family history of asthma in one or more first-degree relatives was consistently identified as a risk factor for asthma. In ten studies, sensitivity and predictive value of a positive family history of asthma could be calculated: sensitivity ranged from 4% to 43%, positive predictive value from 11% to 37%, and negative predictive value from 86% to 97%. Although a positive family history predicts an increased risk of asthma, it identifies a minority of children at risk. Positive family history may have utility in targeting some individual prevention efforts, but the low positive predictive value limits its value as a means to direct environmental remediation efforts. PMID- 12568823 TI - Family history of colon cancer: what does it mean and how is it useful? AB - BACKGROUND: Family history of colon cancer can be deconstructed into causal and noncausal explanations, which include genetic factors, environmental factors, gene-environment interactions, misclassification, and differences in screening. METHODS: We investigated some of these causal and noncausal explanations by using data from a case-control study of colon cancer conducted among African Americans and whites in North Carolina. We examined the relationship between family history and polymorphisms in four genes (N-acetyltransferase 1 and 2 [NAT1, NAT2], methylenetetrahydrofolate reductase, and peroxisome proliferator-activated receptor gamma [PPARG]), environmental risk factors, the joint distributions of these genes and environmental risk factors, and the prevalence of colon cancer screening. RESULTS: Participants with one or more first-degree relatives with colon cancer showed a slightly higher prevalence of at-risk genotypes for each locus, but results were statistically significant only for NAT2. Participants with a family history showed a higher prevalence of at-risk combinations of genotypes and environmental risk factors (NAT2 and well-done red meat consumption; PPARG and nonsteroidal anti-inflammatory medication use). The sensitivity and predictive value of family history for identifying persons with at-risk genotypes or environmental risk factors was low. History of cancer screening was similar in those with and without a family history. CONCLUSIONS: Our results suggest that family history of colon cancer may represent aggregation of some genetic polymorphisms and environmental risk factors. Although it is premature to use family history as a screening tool when testing for genetic polymorphisms, further research is needed to identify additional genes and environmental factors that may be associated with family history. PMID- 12568824 TI - Achieving utility with family history: colorectal cancer risk. AB - Family history of chronic disease is rapidly becoming a research tool for targeting participants at increased risk. Its current usefulness in clinical practice remains unknown. This paper details the possible utility and complications in using family history in a primary care setting, using colorectal cancer risk as the health issue. Where available, we cite data to support the issues that could arise. Where there are no studies, we invite further research. The potential of family history as a health improvement tool is still under review. PMID- 12568825 TI - Effecting behavior change: awareness of family history. AB - This article explores the use of family history of disease as a public health tool for risk stratification and improved disease prevention by drawing from previous research on women at moderate risk of developing breast cancer because of a positive family history. About one quarter to one third of women do not appear to be aware of the added risk a family history of breast cancer poses, and many women with a family history overestimate their risk. It is unclear whether risk perceptions are causally related to breast cancer screening in women with a family history. Exaggerated risk perceptions may not hinder breast cancer screening, unless accompanied by distress. Studies suggest that counseling women with a family history of breast cancer about their risk has a small and short term effect on risk comprehension, a small effect on breast cancer screening, psychological benefits for some women, and unintended negative effects on screening for other women. Future research needs to consider the psychological, individual difference, and cultural variables that moderate counseling effects, recruitment biases, the prospective relationship between perceived risk and breast cancer screening, and whether risk perceptions and comprehension need to match objective risk to be an effective tool to promote screening. PMID- 12568826 TI - Validation of family history data in cancer family registries. AB - BACKGROUND: Although family history information on cancer is used to infer risk of the disease in population-based, case-control, cohort, or family-based studies, little information is available on the accuracy of a proband's report. In this study, we sought to determine the validity of the reporting of family history of cancer by probands in population-based and clinic-based family registries of breast, ovarian, and colorectal cancers. METHODS: To assess the accuracy of probands' reported family history of cancer in their relatives, we compared the family history from the personal interview of each proband to a reference standard that included pathology reports, self-reports, or death certificates on the relatives. Our study included 1111 families that accounted for 3222 relatives who were verified. To account for within-family correlations in the responses, we used a generalized estimating equation approach. RESULTS: The probability of agreement between the proband-reported cancer status in a relative with the reference standard varied by cancer site and by degree of relationship to the proband. This probability for first-degree relatives was 95.4% (95% confidence interval [CI]=92.6-98.3) for female breast cancer; 83.3% (95% CI=72.8-93.8) for ovarian cancer; 89.7% (95% CI=85.4-94.0) for colorectal cancer; and 79.3% (95% CI=70.0-88.6) for prostate cancer. CONCLUSIONS: We found high reliability of probands' reporting on most cancer sites when they reported on first-degree relatives and moderate reliability for their reporting on second- and third-degree relatives. Overreporting of cancer was rare (2.4%). Race or ethnicity and gender of the proband did not influence the accuracy of reporting. However, degree of relationship to the proband, type of cancer, age at diagnosis of the proband, and source of ascertainment of probands were statistically significant predictors of accuracy of reporting. PMID- 12568827 TI - Using decision analytic methods to assess the utility of family history tools. AB - Family history may be a useful tool for identifying people at increased risk of disease and for developing targeted interventions for individuals at higher-than average risk. This article addresses the issue of how to examine the utility of a family history tool for public health and preventive medicine. We propose the use of a decision analytic framework for the assessment of a family history tool and outline the major elements of a decision analytic approach, including analytic perspective, costs, outcome measurements, and data needed to assess the value of a family history tool. We describe the use of sensitivity analysis to address uncertainty in parameter values and imperfect information. To illustrate the use of decision analytic methods to assess the value of family history, we present an example analysis based on using family history of colorectal cancer to improve rates of colorectal cancer screening. PMID- 12568828 TI - Prevention of premature births, a national priority. PMID- 12568829 TI - Modern obstetrical and infertility care may increase the prevalence of disease: an evolutionary concept. AB - OBJECTIVE: To explore the hypothesis that the increasing prevalence of some polygenetically inherited conditions, may, to a degree, be the consequence of treatment successes of modern health care that have overcome evolutionary blocks to reproductive success. DESIGN: Hypothesis, based on a selective review of the literature. SETTING: University-affiliated private infertility center. Patient(s)None. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Occurrence of polygenetically inherited diseases. RESULT(S): Successfully treated evolutionary blocks to the inheritance of polygenetically inherited diseases result in an increased prevalence of these diseases in subsequent generations. CONCLUSION(S): Evolutionary processes have erected barriers to successful reproduction for many polygenetically inherited diseases, which are now overcome by successful treatment, leading to an increased prevalence, younger age of occurrence, and greater severity of these diseases in next generation offspring. PMID- 12568830 TI - Use of luteinizing hormone in the treatment of infertility: time for reassessment? PMID- 12568831 TI - Effects of estrogen and psychological stress on plasma homocysteine levels. AB - OBJECTIVE: To investigate the effects of estrogen (E) and psychological stress on plasma total homocysteine levels in relation to menopausal status. DESIGN: Double blind, randomized, placebo-controlled study. SETTING: The General Clinical Research Center of a university hospital. PATIENT(S): Thirty-six postmenopausal women and 26 premenopausal women. Both samples were healthy nonsmokers. INTERVENTION(S): Both premenopausal and postmenopausal women were subjected to a 6-minute psychological stressor. Postmenopausal women were randomized to one of three treatment arms: 2 mg of E2 or 2 mg of E2 + 5 mg of medroxyprogesterone acetate (MPA), or a placebo, all of which were given orally for 3 months. The psychological stressor was readministered after the 3-month regimen. MAIN OUTCOME MEASURE(S): Plasma total homocysteine levels were measured before and after the psychological stressor on one occasion for premenopausal women and before and after hormone replacement or placebo for postmenopausal women. RESULT(S): There were no significant differences in homocysteine levels between premenopausal (7.2 +/- 1.7 micromol/L; mean +/- SD) and postmenopausal women (7.9 +/- 2.06; mean +/- SD). There was no effect of stress or hormone replacement on homocysteine levels. CONCLUSION(S): Psychological stress, menopausal status, and oral hormone replacement therapy (HRT) do not affect plasma total homocysteine levels in women with normal basal homocysteine levels. PMID- 12568832 TI - A randomized placebo-controlled study of the effect of transdermal vs. oral estradiol with or without gestodene on homocysteine levels. AB - OBJECTIVE: To assess the effect of transdermal vs. oral administration of E2 on plasma homocysteine levels and to evaluate the impact of adding a progestogen to these regimens. DESIGN: Prospective, double-blind, double-dummy, placebo controlled study. SETTING: Outpatient clinics in two university hospitals and two teaching hospitals in The Netherlands. PATIENT(S): One hundred fifty-two healthy hysterectomized postmenopausal women. INTERVENTION(S): Thirteen 28-day treatment cycles with placebo (n = 49); transdermal 17beta-E2, 50 microg (n = 33), oral E2, 1 mg (n = 37), or oral E2, 1 mg, plus gestodene, 25 microg (n = 33), followed by four cycles of placebo in each group. MAIN OUTCOME MEASURE(S): Fasting plasma total homocysteine concentrations at baseline and cycle 4, 13, and 17. RESULT(S): Mean (+/-SD) homocysteine concentrations in the oral E2 group decreased from baseline to cycle 4 (9.0 +/- 2.5 micromol/L vs. 8.2 +/- 2.0 micromol/L; mean change, -7.6%). Homocystine values in the oral E2 plus gestodene group did not change substantially from baseline to cycle 4 (8.9 +/- 1.6 micromol/L vs. 8.6 +/- 2.0 micromol/L; mean change, -4.4%). No significant changes were observed in the transdermal E2 group. After four washout cycles, the homocysteine concentration had returned to baseline values in all groups. CONCLUSION(S): Oral E2 therapy reduced the homocysteine concentration more than did therapy with transdermal E2 or oral E2 plus gestodene. This finding may indicate a role of liver metabolism and suggests that gestodene has a negative effect on these changes. PMID- 12568833 TI - Effect of tibolone treatment on intima-media thickness and the resistive indices of the carotid arteries. AB - OBJECTIVE: To assess the effect of tibolone treatment on the intima-media thickness (IMT) of the common carotid artery (CCA) and the resistive indices (RIs) of the CCA and internal (ICA) and external (ECA) carotid and the vertebral arteries (VAs) in postmenopausal women as sonographic markers of atherosclerosis. DESIGN: A prospective study. SETTING: University hospital. PATIENT(S): Twenty postmenopausal women who had no cardiovascular disease. INTERVENTION(S): Noninvasive measurements of the IMT of the CCA and the RI of the CCA, ICA, ECA, and VA were made with ultrasound at baseline and after 12 weeks of tibolone treatment. MAIN OUTCOME MEASURE(S): IMT of the CCA and RI of the CCA, ICA, ECA, and VA. RESULT(S): Three months of tibolone treatment decreased the IMT of the CCA (mean +/- SD) from 0.70 +/- 0.22 mm (95% confidence interval [CI], 0.60-0.80) to 0.47 +/- 0.17 mm (95% CI, 0.39-0.55) by 28%. Resistive indices of the CCA, ICA, and VA also decreased significantly. CONCLUSION(S): The present study showed that tibolone treatment decreases both the IMT of the CCA and RI of the CCA, ICA, and VA, which appears to be related to its anti-atherosclerotic effect. Nevertheless, the clinical implications of these findings are yet to be investigated. PMID- 12568834 TI - Hormone replacement therapy in postmenopausal women with Alzheimer's disease: a randomized, prospective study. AB - OBJECTIVE: To compare the therapeutic efficacy of hormone replacement therapy (HRT) and tacrine in Alzheimer's disease. DESIGN: Six-month, randomized, open label study. SETTING: University hospital. PATIENT(S): Fifty-five women with mild to moderate Alzheimer's disease were randomly assigned to tacrine (n = 26) or HRT (n = 29). INTERVENTION(S): In the tacrine group, an initial dose of 40 mg/day was increased up to 160 mg/day. In the HRT group, conjugated equine estrogen was given to patients without uteri (n = 3) or together with micronized progesterone to patients with uteri (n = 26). MAIN OUTCOME MEASURE(S): Mini-Mental State Examination (MMSE), Hopkins Verbal Learning Test, Boston Naming Test, Controlled Oral Word Association Test, Geriatric Depression Scale, Hamilton Depression Scale (HDS), and Instrumental Activities of Daily Living (IADL). RESULT(S): Thirty three patients who completed the outcome measures (tacrine, 17; HRT, 16) were included in an intent-to-treat analysis. The results did not differ between groups except for IADL, which rated more highly after HRT. Apolipoprotein E genotype effects were assessed. MMSE and HDS scores were improved after tacrine treatment in epsilon 4-negative patients. CONCLUSION(S): Overall efficacy of estrogen plus progesterone combination was similar to tacrine for cognition and mood, but greater for ADL. In epsilon 4-negative patients, tacrine is preferable for cognition and mood. PMID- 12568835 TI - Clinical and endocrine follow-up of patients after testicular sperm extraction. AB - OBJECTIVE: To evaluate the risk of testicular damage from testicular biopsies that are carried out for testicular sperm extraction (TESE) in infertile men. DESIGN: Prospective controlled clinical study. SETTING: Academic hospital. PATIENT(S): Forty infertile males with azoospermia. Examination of the clinical, endocrine, biochemical, and sonographic data in average after 18 months after TESE was performed. MAIN OUTCOME MEASURE(S): Measurements before and after TESE: hormone values, testicular size, morphologic characteristics, and power Doppler after scrotal sonography. RESULT(S): Comparison of preoperative and postoperative values of basal testosterone, FSH, LH, and estradiol levels did not reveal any differences. Twelve of 26 patients had subnormal testosterone values before TESE; 14 of 39 patients had subnormal levels afterward. Postoperative sonographic measurements showed no significant difference of the testicular volume as compared with the preoperative values. Results of power Doppler sonography revealed pathological conditions (n = 5) in patients with former iliacal or testicular operations. CONCLUSION(S): Endocrine testicular function and testicular size were not impaired after testicular biopsy when compared with preoperative data. However, patients with nonobstructive azoospermia seem to be at risk for androgen deficiency due to primary testicular failure after repeated testicular biopsies. PMID- 12568836 TI - Semen quality of male smokers and nonsmokers in infertile couples. AB - OBJECTIVE: To investigate the effect of cigarette smoking on main sperm variables. DESIGN: Cohort study. SETTING: Men attending the andrology laboratory in the context of infertility investigation in the couple. PATIENT(S): Eight hundred thirty-nine smokers and 1,266 non-smokers were enrolled; 655 smokers and 1,131 nonsmokers fulfilled the inclusion criteria and were compared. Standard clinical analysis of semen. RESULT(S): Cigarette smoking was associated with a significant decrease in sperm density (-15.3%), total sperm count (-17.5%), total number of motile sperm (-16.6%), and citrate concentration (-22.4%). The percentage of normal forms was significantly reduced in smokers, and sperm vitality, ejaculate volume, and fructose concentration were slightly but nonsignificantly affected. CONCLUSION(S): Cigarette smoking is associated with reduced semen quality. PMID- 12568837 TI - Use of carnitine therapy in selected cases of male factor infertility: a double blind crossover trial. AB - OBJECTIVE: To determine the efficacy of L-carnitine therapy in selected cases of male factor infertility. DESIGN: Placebo-controlled, double-blind, crossover trial. SETTING: University tertiary referral center. PATIENT(S): One hundred infertile patients (ages 20-40 years) with the following baseline sperm selection criteria: concentration, 10-20 x 10(6)/mL; total motility, 10%-30%; forward motility, <15%; atypical forms, <70%; velocity, 10-30 micro/s; linearity, <4. Eighty-six patients completed the study. INTERVENTION(S): Patients underwent L carnitine therapy 2 g/day or placebo; the study design was 2 months of washout, 2 months of therapy/placebo, 2 months of washout, and 2 months placebo/therapy. MAIN OUTCOME MEASURE(S): Variation in sperm parameters used in the patients selection criteria, in particular, sperm motility. Excluding outliers, a statistically significant improvement in semen quality, greater than after the placebo cycle, was seen after the L-carnitine therapy for sperm concentration and total and forward sperm motility. The increase in forward sperm motility was more significant in those patients with lower initial values, i.e., <5 x 10(6) or <2 x 10(6) of forward motile sperm/ejaculate or sperm/mL. CONCLUSION(S): Based on a controlled study of efficacy, L-carnitine therapy was effective in increasing semen quality, especially in groups with lower baseline levels. However, these results need to be confirmed by larger clinical trials and in vitro studies. PMID- 12568838 TI - Cytogenetic and Y chromosome microdeletion screening of a random group of infertile males. AB - OBJECTIVE: To assess whether to perform routine cytogenetic and Y chromosome microdeletion screening on all infertile male patients. DESIGN: A cytogenetic and Y microdeletion study of a random group of infertile men. SETTING: University department. PATIENT(S): In total, 40 patients had azoospermia (21 nonidiopathic), 27 had severe oligozoospermia/oligoasthenozoospermia (10 IU/L, polycystic ovarian syndrome, one ovary or previous ovarian surgery, previous poor response to ovarian stimulation, or ovarian hyperstimulation syndrome (OHSS). INTERVENTION(S): Randomized to commence 150 IU or 225 IU of Gonal-F per day without dose alterations during treatment. MAIN OUTCOME MEASURE(S): Number of oocytes retrieved and total rFSH dose. RESULT(S): More oocytes were retrieved in women aged or=33 years), the number of oocytes retrieved in the two groups were similar. No significant differences were found for fertilization rate, number of embryos formed and cryopreserved, and pregnancy rates between the two groups. The total rFSH dose used was higher in the 225-IU group (2,595.0 +/- 510.0 vs. 1,897.5 +/- 457.5 IU). The cancellation rate due to insufficient ovarian response was higher in the 150-IU group (15.0% vs. 3.3%). All cases of ovarian hyperstimulation syndrome (n = 4) occurred in the 225-IU group. CONCLUSION(S): Two hundred twenty-five IU is more effective than 150 IU in younger women but requires a higher total dose of Gonal-F. The use of 225 IU in older women did not result in a higher oocyte yield, suggesting that 225 IU of rFSH does not compensate for the age-related decline in the number of follicles available for stimulation. PMID- 12568840 TI - Coordination of early antral follicles by luteal estradiol administration provides a basis for alternative controlled ovarian hyperstimulation regimens. AB - OBJECTIVE: To investigate whether luteal E(2) administration reduces size discrepancies of early antral follicles. DESIGN: Prospective, crossover study. SETTING: ART unit, Clamart, France. PATIENT(S): Sixty women and 120 cycles. INTERVENTION(S): On cycle day 3 (baseline day 3), all women underwent measurements of early antral follicles by ultrasound and serum FSH and ovarian hormones. From day 20 until the next cycle day 2, 30 of them received oral 17beta E(2), whereas the remaining women served as controls. The day after E(2) discontinuation (E(2) day 3) or on subsequent cycle day 3 (control day 3), participants were reevaluated as on baseline day 3. MAIN OUTCOME MEASURE(S): Magnitude of follicular size discrepancies. RESULT(S): Follicular size discrepancies and follicular diameters were significantly attenuated on E(2) day 3 (3.7 +/- 0.5 mm) as compared with baseline day 3 (4.9 +/- 1.0 mm), but not in controls (5.0 +/- 0.8 vs. 4.9 +/- 0.8 mm). FSH (4.3 +/- 1.9 vs. 7.3 +/- 3.3 mIU/mL) and inhibin B (34 +/- 28 vs. 71 +/- 32 pg/mL) levels were consistently lower on E(2) day 3 than on baseline day 3 but remained unchanged in controls. CONCLUSION(S): Luteal E(2) administration reduces the size and improves the homogeneity of early antral follicles on day 3. This approach may be instrumental in synchronizing follicular development during controlled ovarian hyperstimulation. PMID- 12568841 TI - Bleeding patterns after immediate vs. conventional oral contraceptive initiation: a randomized, controlled trial. AB - OBJECTIVE: To compare bleeding patterns after immediate vs. conventional oral contraceptive (OC) initiation. DESIGN: Randomized controlled trial. SETTING: University-based clinic. PATIENT(S): One hundred thirteen women initiating combination OCs. INTERVENTION(S): Participants received a 4-month supply of a monophasic 35-microg ethinyl E(2) (EE) OC and a bleeding diary, were randomized to immediate or conventional OC start, underwent monthly telephone follow-up, and after 90 days returned the diary and completed an exit interview. MAIN OUTCOME MEASURE(S): Total number of bleeding-spotting days, using the World Health Organization 90-day reference period method. Comparisons were made by trial assignment (immediate vs. conventional) and cycle day of OC initiation (day 8+ vs. days 1-7). RESULT(S): There was no significant difference in the number of bleeding-spotting days (mean difference: -0.5 days; 95% CI: -3.4 to 2.3) or any other bleeding parameter between the immediate and conventional starters, or days 1-7 and day 8+ starters. CONCLUSION(S): Immediate start of OCs does not induce bleeding patterns different from conventional starting regimens. Concern about adverse bleeding patterns should not be considered a justification for instructing women to wait until menses before starting OCs. PMID- 12568842 TI - Effects of maternal and paternal alcohol consumption on the success rates of in vitro fertilization and gamete intrafallopian transfer. AB - OBJECTIVE: To determine whether the amount and timing of female and male alcohol use during IVF and GIFT affect reproductive endpoints. DESIGN: Multicenter prospective study. SETTING: Clinics in southern California. PATIENT(S): Two hundred twenty-one couples with female infertility. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Egg retrieval, transfer, fertilization, pregnancy, miscarriage, live birth, and multiple gestations. RESULT(S): Female alcohol consumption was associated with: (1) a 13% decrease in the number of eggs aspirated (adjusted 95% confidence interval [CI]: -2% to -23%, for one additional drink per day, 1 year before the IVF or GIFT attempt); (2) an increase in risk of not achieving pregnancy by 2.86 times (0.99-8.24, 1 month prior); and (3) an increase in risk of miscarriage by 2.21 times (1.09-4.49, 1 week before the procedure). For men, one additional drink per day increased the risk of not achieving a live birth by 2.28 (1.08-4.80) to 8.32 (1.82-37.97) times, depending on the time period; beer also affected live births (ORs = 5.49-45.64). This outcome may be due partially to increased risk of miscarriage by 2.70 to 38.04 times for men who drank 2.5, free T 5.6 pg/mL, DHEAS 2,070 ng/mL, androstenedione (A) 3.42 ng/mL; whereas 17-beta-E(2), P, PRL, and free T(4) values were within the normal range. Ultrasonography demonstrated a typical polycystic echographic architecture of the ovaries. CONCLUSION(S): Lipodystrophy and insulin resistance are well-recognized side effects of HAART. Polycystic ovary syndrome might be an additional side effect secondary to insulin resistance and lipodystrophy. PMID- 12568858 TI - Dizygotic twin delivery following in vitro fertilization and transfer of thawed blastocysts cryopreserved at day 6 and 7. AB - OBJECTIVE: To report the first conception and delivery following transfer of thawed human blastocysts maintained in extended in vitro culture with cryopreservation at day 6 and 7. DESIGN: Case report. SETTING: Major urban infertility referral center. PATIENT(S): A 26-year-old woman with pelvic endometriosis and two prior unsuccessful in vitro fertilization/embryo transfer (IVF-ET) attempts. INTERVENTION(S): The patient underwent controlled ovarian hyperstimulation using a combined FSH + hMG protocol, and 24 oocytes were retrieved. MAIN OUTCOME MEASURE(S): Dizygotic twin delivery after IVF and intracytoplasmic sperm injection (ICSI), assisted embryo hatching, and ultrasound guided transfer of cryopreserved blastocysts. RESULT(S): After three embryos were subjected to assisted hatching, they were transferred fresh on day 3, but no implantation occurred. All nontransferred embryos (n = 11) were observed during extended in vitro culture and three blastocysts were selected for cryopreservation on day 6 and 7; thaw and transfer occurred the following month and a pregnancy was achieved. Dizygotic twins (female/female) were delivered by cesarean in the early third trimester. CONCLUSION(S): Substantial advancements have been made in the field of embryo cryogenics and in vitro fertilization, but controversy remains regarding the value of freezing late-developing human blastocysts. Here we describe the first reported live births with IVF after extended in vitro culture and cryopreservation at day 6 and 7 after fertilization. PMID- 12568859 TI - Successful resection of a heterotopic cervical pregnancy resulting from intracytoplasmic sperm injection. AB - OBJECTIVE: To report a case of cervical heterotopic pregnancy after IVF-ET treatment. DESIGN: Case report. SETTING: Private IVF center. PATIENT(S): A woman who had undergone controlled ovarian hyperstimulation and intracytoplasmic sperm injection-ET for primary male factor infertility. INTERVENTION(S): Resection of cervical heterotopic pregnancy by hysteroscopy. MAIN OUTCOME MEASURE(S): Successful treatment of heterotopic cervical pregnancy. RESULT(S): A successful pregnancy resulting in a term baby. CONCLUSION(S): Hysteroscopic resection can be used to treat a cervical heterotopic pregnancy. PMID- 12568860 TI - Fertility potential of women with congenital ampullary atresia of the fallopian tube. AB - OBJECTIVE: To determine the fertility potential of women with congenital ampullary atresia of the fallopian tube. DESIGN: Case report and review of literature. SETTING: University-affiliated tertiary care infertility clinic. PATIENT(S): Six infertile women with congenital ampullary atresia of the oviduct. INTERVENTION(S): Salpingostomy and fimbrial approximation. MAIN OUTCOME MEASURE(S): Pregnancy. RESULT(S): Of six reported cases of congenital ampullary atresia of the fallopian tube who underwent surgery, four women conceived intrauterine pregnancies (IUP) and another, a pregnancy of undetermined site. CONCLUSION(S): Salpingostomy and fimbrial approximation is a therapeutic option for infertile women with congenital ampullary atresia of the fallopian tube. PMID- 12568861 TI - Isolated follicle-stimulating hormone (FSH) deficiency in a young man with normal virilization who did not have mutations in the FSHbeta gene. AB - OBJECTIVE: To determine the cause of isolated FSH deficiency in a young infertile man. DESIGN: Case report. SETTING: Clinical and genetic studies in an academic research environment. PATIENT(S): A 19-year-old man with normal virilization, azoospermia, and isolated FSH deficiency. INTERVENTION(S): Pituitary and gonadal functions were evaluated at baseline and after repeated GnRH stimulation. FSH was tested with both immunological and biological methods. The FSHbeta gene was sequenced in the patient and in a series of 50 controls. MAIN OUTCOME MEASURE(S): Clinical, endocrine, and genetic characterization of an infertile patient with isolated FSH deficiency. RESULT(S): LH and T secretions were normal. No interference in FSH measurement was detected, and serum FSH concentrations were very low and completely unresponsive to repeated GnRH stimulation. No circulating FSH-like bioactivity was detected by means of rat Sertoli cell bioassay. Other pituitary functions were unaffected, and no lesions were seen at pituitary nuclear magnetic resonance (NMR). Inhibin B and activin levels were normal, but a progressive decrease of activin concentrations was seen during GnRH stimulation. The coding sequence of the FSHbeta gene was normal, but the patient was homozygous for a novel G/T substitution in the promoter region within a P response element. This substitution was present in heterozygosity in eight out of 50 controls and in homozygosity in one man with normal FSH levels. CONCLUSION(S): We report an infertile male with isolated FSH deficiency but no evidence of mutations in the FSHbeta gene. The G/T substitution in the FSHbeta promoter represents a novel silent polymorphism, indicating that other defects in factors involved in FSH-specific expression should be taken into account. PMID- 12568862 TI - Adnexal torsion in a patient with hydrosalpinx who underwent tubal occlusion before in vitro fertilization. AB - OBJECTIVE: To report a rare case of adnexal torsion in an infertile patient with bilateral hydrosalpinges who had proximal tubal occlusion performed before IVF-ET 5 years earlier. DESIGN: A case report and literature review. SETTING: Reproductive endocrine division in a community teaching hospital. PATIENT(S): A 43-year-old infertile woman with acute left-side pelvic pain. INTERVENTION(S): Laparoscopic bilateral salpingectomies. MAIN OUTCOME MEASURE(S): Diagnosis and etiology of pelvic pain with subsequent therapy to alleviate symptoms. RESULT(S): The patient experienced resolution of her pain and prophylactic prevention of further torsion status after bilateral salpingectomies. CONCLUSION(S): Although laparoscopic proximal tubal occlusion is an effective treatment of hydrosalpinges in patients undergoing IVF-ET, it carries the risk of torsion of the hydrosalpinx and the associated morbidities. PMID- 12568863 TI - Successful pregnancies after combined pentoxifylline-tocopherol treatment in women with premature ovarian failure who are resistant to hormone replacement therapy. AB - OBJECTIVE: To determine whether treatment with combined pentoxifylline (PTX) and tocopherol (Vit.E) can improve uterine parameters in hormonal replacement therapy (HRT)-resistant women with premature ovarian failure (POF), for whom the outcome of assisted reproductive technology is usually negative. We previously reported that uterine radiation-induced fibrosis is reversible by combined PTX-Vit.E treatment. DESIGN: Case report. SETTING: Volunteer participants in an oocyte donation (OD) program in a French public hospital. PATIENT(S): Three women with POF (ages 36 +/- 2 years) using HRT exhibited uterine hormonoresistance, although they had high E(2) plasma levels. Their mean endometrial thickness was 4.9 mm, and they had an echogenic endometrium and thin uterine crosses. INTERVENTION(S): Between May 1998 and April 1999, treatment consisted of 800 mg of PTX combined with 1,000 IU of Vit.E daily for at least 9 months. MAIN OUTCOME MEASURE(S): Endometrial thickness, echogenicity, and pulsatility index of the uterine arteries, assessed by ultrasound and Doppler before and after treatment, and embryo implantation by IVF-OD. RESULT(S): PTX-Vit.E treatment was well tolerated and induced improvements, as mean edematous endometrial thickness increased to 7.4 mm, with nice uterine crosses. Three frozen-thawed ETs resulted in two viable pregnancies. CONCLUSION(S): In women with POF and uterine resistance to HRT, combined PTX-Vit.E reduces fibroatrophic uterine lesions and improves the uterine response to HRT, thus allowing embryo implantation and ongoing pregnancy. PMID- 12568864 TI - A case of complete hypogonadotropic hypogonadism with a mutation in the gonadotropin-releasing hormone receptor gene. AB - OBJECTIVE: To screen for mutations in the GnRH receptor gene in a case of complete hypogonadotropic hypogonadism (HH) with GnRH resistance. DESIGN: Case report. SETTING: A university hospital. PATIENT(S): A male patient with the complete form of HH without anosmia. INTERVENTION(S): Physical examination and laboratory and genetic studies. MAIN OUTCOME MEASURE(S): Gonadotropins at the basal state and after GnRH administration and GnRH receptor DNA sequencing. RESULT(S): A novel missense mutation, localized in the first amino acid of the extracellular loop found in the heterozygous state, and another mutation, Arg(139)His (R139H), located in the conserved aspartate-arginine-serine motif at the junction of the third transmembrane and second intracellular loop of the GnRH receptor, were identified in the homozygous state. Pedigree studies reveal that both parents were heterozygous for R139H, while the mother carried the missense mutation at codon 1(M1T). CONCLUSION(S): GnRH receptor mutations may account for a larger proportion of cases of HH than previously thought. The phenotypic spectrum of HH seems to vary, and this heterogeneity may be related, at least in part, to the degree of impaired biological activity of the mutated GnRH receptor caused by the allelic type of mutations. PMID- 12568865 TI - Mutation analysis of BrCA1, BrCA2, and p53 versus soluble HLA class I and class II in a case of familial endometriosis. AB - OBJECTIVE: To investigate possible correlation(s) between mutations of BrCA1, BrCA2, and p53 genes versus soluble HLA expression in familial endometriosis. DESIGN: Mutation analysis. SETTING: University teaching departments and hospital. PATIENT(S): A family with seven women in two generations with familial endometriosis. INTERVENTION(S): Mutation analysis of BrCA1, BrCA2, and p53 genes. MAIN OUTCOME MEASURE(S): A point mutation of the BrCA1 gene appears to inhibit soluble HLA secretion. RESULT(S): Among the three genes examined, only the BrCA1 gene showed a T to A mutation at position 3232 that correlates with total abolishment of both class I and class II antigen release. CONCLUSION(S): A possible correlation between a BrCA1 mutation and soluble HLA expression appears to exist. The mutation is not stage dependent and seemingly influences the secretion of both class I and class II antigens that are totally absent from the serum of only one family member. PMID- 12568866 TI - The origin of biochemical pregnancies: lessons learned from preimplantation genetic diagnosis. PMID- 12568868 TI - Developmental monitoring of children conceived by intracytoplasmic sperm injection and in vitro fertilization. PMID- 12568869 TI - Effect of raloxifene, estrogen, and hormone replacement therapy on serum homocysteine levels in postmenopausal women. PMID- 12568867 TI - Ovarian brain-derived neurotrophic factor is present in follicular fluid from normally cycling women. PMID- 12568870 TI - Antisperm antibodies in hydroceles before and after surgery. PMID- 12568871 TI - Profound luteinizing hormone suppression induces a deleterious follicular environment during assisted reproduction technology. PMID- 12568872 TI - In syndromes of confused gonads, height can be important! PMID- 12568873 TI - In syndromes of confused gonads, height can be important! PMID- 12568875 TI - In syndromes of confused gonads, height can be important! PMID- 12568909 TI - Baicalein inhibits Raf-1-mediated phosphorylation of MEK-1 in C6 rat glioma cells. AB - Baicalein is a flavonoid derived from the Scutellaria root. In investigations of the inhibitors of prostaglandin synthesis in C6 rat glioma cells, we found that baicalein had a potent inhibitory activity on prostaglandin synthesis induced by either histamine or A23187, a Ca(2+) ionophore. Baicalein inhibited histamine- or A23187-induced phosphorylation of p42/p44 extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK), which causes the phosphorylation of cytosolic phospholipase A(2) (PLA(2)). Baicalein also inhibited the phosphorylation of MAPK kinase-1 (MEK-1) induced by histamine or A23187 in the cells. To examine the site of action of baicalein, MEK-1 and Raf-1 were prepared by immunoprecipitation with anti-MEK-1 and anti-Raf-1 antibodies, respectively. Baicalein inhibited the phosphorylation of exogenous MEK-1 by Raf-1 under cell free conditions, while it did not change the phosphorylation of exogenous p42 MAPK by MEK-1. These results imply that baicalein inhibits the ERK/MAPK cascade, acting on the phosphorylation of MEK-1 by Raf-1. PMID- 12568910 TI - Argatroban, specific thrombin inhibitor, induced phenotype change of cultured rabbit vascular smooth muscle cells. AB - To investigate whether argatroban ((2R,4R)-4-methyl-1-[N(2)-((RS)-3-methyl 1,2,3,4-tetrahydro-8-quinolinesulfonyl)-L-arginyl]-2-piperidinecarboxylic acid hydrate, a selective thrombin inhibitor, exerts a direct action on phenotype conversion of vascular smooth muscle cells, cultured rabbit aortic vascular smooth muscle cells were employed. Myosin heavy chain isoforms (SM1, SM2, and SMemb) mRNA expressions were evaluated by in situ hybridization and reverse transcription-polymerase chain reaction (RT-PCR). After the cells were incubated in serum-free medium containing argatroban (10 and 50 microg/ml) and platelet derived growth factor (PDGF)-BB (10 and 50 ng/ml) for 3 h, total RNA was extracted. In situ hybridization demonstrated that myosin heavy-chain isoform mRNAs were homogenously expressed in argatroban- and PDGF-BB-treated cells. RT PCR revealed that SM1/SM2 mRNA expressions were not changed with argatroban, while SMemb mRNA expression was increased to 1.6-fold with a statistical significance (P<0.05). Treatment with argatroban (10 and 50 microg/ml) at 24 h did not change SM1/SM2 mRNA expressions. Although SMemb mRNA expression was slightly increased, there was no statistical significance. Other phenotype markers including plasminogen activator inhibitor-1 (PAI-1) and beta-actin mRNAs were also significantly increased by argatroban. In conclusion, argatroban can directly induce phenotype conversion of vascular smooth muscle cells with the resultant up-regulation of SMemb, PAI-1, and beta-actin mRNAs. PMID- 12568911 TI - Targeted gene deletion of the 5-HT3A receptor subunit produces an anxiolytic phenotype in mice. AB - Anxiety disorders are the most common psychiatric disorders. Typical medications used to treat patients are benzodiazepines or antidepressants that target serotonin (5-HT) activity. The ionotropic 5-HT(3) receptor has emerged as a potential therapeutic target because selective antagonist compounds reduce anxiety in rodents, primates, and humans. 5-HT binds to the extracellular N terminus of the 5-HT(3A) receptor subunit, but receptor activation is also enhanced by distinct allosteric sites. It is not known if specific molecular subunits of the 5-HT(3) receptor modulate anxiety. To address this issue, we characterized anxiety-like behavior of mice with a targeted deletion of the 5 HT(3A) receptor subunit gene in the light/dark box, elevated plus maze, and novelty interaction animal models of anxiety. 5-HT(3A) null mice exhibited an anxiolytic behavioral phenotype that was highly correlated across behavioral measures. This evidence indicates that the 5-HT(3A) molecular subunit influences anxiety-like behavior. Pharmacotherapy that targets specifically the 5-HT(3A) receptor subunit may provide a novel treatment for anxiety disorders. PMID- 12568912 TI - Milk intake and survival in newborn cannabinoid CB1 receptor knockout mice: evidence for a "CB3" receptor. AB - Cannabinoids, whether plant-derived, synthetic or endogenous, have been shown to stimulate appetite in the adult organism. We have reported previously that cannabinoid receptors play a critical role during the early suckling period: The selective cannabinoid CB(1) receptor antagonist N-(piperidiny-1-yl)-5-(4 chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (SR141617A) permanently prevented milk ingestion in a dose-dependent manner, when administered to (Sabra, albino) mouse pups, within 1 day of birth. As a consequence, these pups died within the first week of life. We now generalize this finding to a different strain of mice (C57BL/6). Further, we show that cannabinoid CB(1) receptor blockade (20 mg/kg SR141716A) must occur within 24 h after birth as injection of SR141716A into 2- or 5-day-old pups had a much smaller effect or no effect at all, respectively. Cannabinoid CB(1) receptor knockout mice did not ingest milk on the first day of life, similarly to SR141716A-treated normal pups, as measured by the appearance of "milkbands". However, the knockout pups started to display milkbands from day 2 of life. Survival rates of cannabinoid CB(1) receptor knockout mice were affected significantly, but to a lesser extent than normal pups, by the administration of SR141716A. Daily administration of the endocannabinoid 2-arachidonoyl glycerol, or the synthetic agonists (R)-(+)-[2,3-dihydro-5-methyl-3-(4 morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone (WIN55,212-2, 5 mg/kg) or (-)-cis-3-[2-Hydroxy4-(1,1-dimethylheptyl) phenyl] trans-4-(3-hydroxypropyl)cyclohexanol (CP55,940, 5 or 20 mg/kg) did not promote survival or weight gain in CB(1)(-/-) pups. Our data support previous evidence for a critical role of cannabinoid CB(1) receptors for the initiation of suckling. Further, the present observations support the existence of an unknown cannabinoid receptor, with partial control over milk ingestion in newborns. Our data also suggest that the CB(1)(-/-) neonates possess a compensatory mechanism which helps them overcome the lack of cannabinoid CB(1) receptors. PMID- 12568913 TI - Blockade of mu-opioid receptor-mediated G-protein activation and antinociception by TRK-820 in mice. AB - The effects of kappa-opioid receptor agonists trans-3,4-dichloro-N-(2-(1 pyrollidinyl)-cyclohexyl) benzeneacetamide ((-)-U50,488H) and 17 cyclopropylmethyl-3,14beta-dihydroxy-4,5alpha-epoxy-6beta-[N-methyl-trans-3-(3 furyl)acrylamido]morphinan hydrochloride (TRK-820) on the G-protein activation and antinociception induced by the selective mu-opioid receptor agonist, [D Ala(2),N-MePhe(4),Gly-ol(5)]enkephalin (DAMGO), were determined in mice. G protein activation was measured by monitoring the guanosine-5'-O-(3 [35S]thio)triphosphate ([35S]GTPgammaS) binding in the mouse pons/medulla. DAMGO (10 microM) produced a marked increase of [35S]GTPgammaS binding to the mouse pons/medulla membrane. On the other hand, both TRK-820 and (-)-U50,488H produced small but significant increases of [35S]GTPgammaS binding to the mouse pons/medulla membrane. These increases by both TRK-820 and (-)-U50,488H were completely reversed by the selective kappa-opioid receptor antagonist, norbinaltorphimine. Under these same conditions, the DAMGO-induced increase of [35S]GTPgammaS binding was significantly attenuated by TRK-820 in a concentration dependent manner, but not by (-)-U50,488H. In the tail-flick test, DAMGO (16 ng) given intracerebroventricularly (i.c.v.), produced a marked antinociception. The antinociception induced by DAMGO was dose-dependently blocked by co-treatment with TRK-820, but not (-)-U50,488H, in mice pretreated with norbinaltorphimine (5 microg, i.c.v.). The present results provide direct evidence for the antagonistic property of TRK-820 for mu-opioid receptors, in addition to the full agonistic property for kappa-opioid receptors. PMID- 12568914 TI - The action of nitric oxide on hepatic haemodynamics during secondary biliary cirrhosis in the rat. AB - The role of nitric oxide (NO) in portal hypertension is poorly understood. The role of NO upon hepatic arterial and portal venous vasoconstrictor responses to noradrenaline and ATP in rats with secondary biliary cirrhosis was evaluated. Cirrhosis was induced by bile duct ligation after which livers were excised and dual-perfused in vitro. Concentration-dependent dose-response curves were then constructed to hepatic arterial and portal venous noradrenaline and ATP. Hepatic arterial responses to noradrenaline and ATP were significantly attenuated in cirrhotic rats. 100 microM N(G)-nitro-L-arginine methyl ester (L-NAME) restored attenuated hepatic arterial responses to noradrenaline and ATP in cirrhotic rats. Portal venous responses to noradrenaline in cirrhotic rats were significantly increased compared to controls and were not affected by L-NAME. However, portal venous responses to ATP were significantly attenuated in cirrhotic rats and were also not restored by L-NAME. Hepatic arterial or portal venous responses to noradrenaline did not change after infusion of L-NAME. Hepatic arterial responses to noradrenaline and ATP were significantly attenuated in cirrhotic rats, possibly due to increased production of NO. However, portal venous responses in cirrhotic rats were increased to noradrenaline and attenuated to ATP, and were not related to increased NO production. PMID- 12568915 TI - SSR69071, an elastase inhibitor, reduces myocardial infarct size following ischemia-reperfusion injury. AB - Neutrophil elastase contributes to the severity of cardiac damage following coronary ischemia and reperfusion. We evaluated the effects of 2-(9-(2 piperidinoethoxy)-4-oxo-4H-pyridol[1,2-a]pyrimidin-2-yloxymethyl)-4-(1 methyethyl)-6-methoxy-1,2-benzisothiazol-3(2H)-one-1,1-dioxide hemihydrate (SSR69071), a novel, potent and selective inhibitor of neutrophil elastase, on infarct size in anaesthetized rabbits subjected to coronary artery occlusion for 30 min followed by reperfusion for 120 min. SSR69071 (3 mg/kg i.v.) reduced cardiac infarct size when administered before ischemia (-39%, P<0.05) or just prior to reperfusion (-37%, P<0.05). Subsequent experiments using the latter administration protocol confirmed the ability of SSR69071 (1 and 3 mg/kg i.v.) to reduce infarct size. This cardioprotective activity was associated with inhibition of cardiac elastase. PMID- 12568916 TI - Assessment of the anti-inflammatory activity and free radical scavenger activity of tiliroside. AB - Three flavonoids, gnaphaliin, pinocembrin and tiliroside, isolated from Helichrysum italicum, were studied in vitro for their antioxidant and/or scavenger properties and in vivo in different models of inflammation. In vitro tests included lipid peroxidation in rat liver microsomes, superoxide radical generation in the xanthine/xanthine oxidase system and the reduction of the stable radical 1,1-diphenyl-2-pycryl-hydrazyl (DPPH). Acute inflammation was induced by application of 12-O-tetradecanoylphorbol 13-acetate (TPA) to the mouse ear or by subcutaneous injection of phospholipase A(2) or serotonin in the mouse paw. Eczema provoked on the mouse ear by repeated administration of TPA was selected as a model of chronic inflammation. The flavonoids were assayed against sheep red blood cell-induced mouse paw oedema as a model of delayed-type hypersensitivity reaction. The most active compound, both in vitro and in vivo, was tiliroside. It significantly inhibited enzymatic and non-enzymatic lipid peroxidation (IC(50)=12.6 and 28 microM, respectively). It had scavenger properties (IC(50)=21.3 microM) and very potent antioxidant activity in the DPPH test (IC(50)=6 microM). In vivo, tiliroside significantly inhibited the mouse paw oedema induced by phospholipase A(2)(ED(50)=35.6 mg/kg) and the mouse ear inflammation induced by TPA (ED(50)=357 microg/ear). Pinocembrin was the only flavonoid that exhibited anti-inflammatory activity in the sheep red blood cell induced delayed-type hypersensitivity reaction. However, only tiliroside significantly reduced the oedema and leukocyte infiltration induced by TPA. As in the case of other flavonoids, the anti-inflammatory activity of tiliroside could be based on its antioxidant properties, although other mechanisms are probably involved. PMID- 12568918 TI - CP8668, a novel orally active nonsteroidal progesterone receptor modulator with tetrahydrobenzindolone skeleton. AB - We investigated progestational activity of a new nonsteroidal compound, CP8668, ((4aR,5R,6R,7R)-7-methoxy-6-(N-propylaminocarbonyl)oxy-4a,5,6,7-tetrahydro 1,3,4a,5-tetramethylbenz[f]indol-2(4H)-one). CP8668 showed selective affinity for human progesterone receptor equal in strength to other steroidal progestins. CP8668 showed no significant affinity for human glucocorticoid receptor or human estrogen receptor and very weak affinity for rat androgen receptor. In endogenous and exogenous progesterone-dependent enzyme expression assays using human mammary carcinoma T47D, CP8668 showed mixed agonist-antagonist activity. However, in a rabbit endometrial transformation test, CP8668 showed good progestational activity following s.c. and p.o. administration. These results suggest that CP8668 is a selective and orally active progesterone receptor modulator, which shows mixed agonist-antagonist activity in in vitro transcription tests and agonist activity in endometrial transformation assays in rabbits, and that it is potentially a promising lead compound for a new type of orally active progesterone receptor modulator. PMID- 12568917 TI - Role of bacteria and inducible nitric oxide synthase activity in the systemic inflammatory microvascular response provoked by indomethacin in the rat. AB - The role of bacteria and nitric oxide (NO), formed by the inducible isoform of NO synthase (iNOS), in a widespread systemic inflammatory microvascular response that follows indomethacin administration, has been investigated in the rat. Subcutaneous administration of indomethacin (10 mg kg(-1)) daily for 2 days produced an increase in microvascular leakage of radiolabelled albumin accompanied by expression of iNOS activity in the lung, liver, spleen and kidney, as well as in the jejunum, caecum, colon and ileum. Pretreatment with dexamethasone (1 mg kg(-1) day(-1), s.c.) reduced indomethacin-provoked microvascular leakage and the expression of iNOS activity in all the tissues studied. The widespread microvascular leakage and iNOS activity was also inhibited by pretreatment with ampicillin (200 mg kg(-1) day(-1), p.o.), metronidazole (200 mg kg(-1) day(-1), p.o.) or by polymyxin B (15 mg kg(-1) day( 1), s.c.). Administration of the highly selective iNOS inhibitor GW 273629 (3-[[2 (ethanimidoylamino)ethyl]sulphonyl]-L-alanine; five doses of 5 mg kg(-1), s.c. over 48 h) substantially inhibited the microvascular leakage in the affected organs. Such findings suggest the involvement of indigenous gut bacteria, lipopolysaccharide and iNOS expression following indomethacin-induced enteropathy in this widespread systemic inflammatory microvascular response. PMID- 12568919 TI - Synthesis of starch derivatives with labile cationic groups. AB - A new route to starch derivatives bearing hydrolyzable cationic groups was developed. This was based on reacting starch compounds with betaine derivatives in the presence of diisopropylcarbodiimide and 4-dimethylaminopyridine as coupling reagents in an aprotic polar solvent. Water-soluble starches with a perfectly controlled degree of substitution were thus obtained which were fully characterized by infrared, 1H and 13C spectroscopy and viscosity measurements. The cationic groups grafted on the polysaccharides are shown to hydrolyze slowly upon storage at room temperature. PMID- 12568920 TI - Effect of a modified thymine on the structure and stability of [d(TGGGT)]4 quadruplex. AB - Telomeric guanine-rich sequence can adopt quadruplex structures that are important for their biological role in chromosomal stabilisation. G quartets are characterised by the cyclic hydrogen bonding of four guanine bases in a coplanar arrangement and their stability is ion-dependent. In this work we compare the stability of [d(TGGGT)](4) and [d(T*GGGT)](4) quadruplexes. The last one contains a modified thymine, where the hydroxyl group substitutes one hydrogen atom of the methyl group of the thymine in the [d(TGGGT)](4) sequence. We used a combination of spectroscopic, calorimetric and computational techniques to characterise the G quadruplex formation. NMR and CD spectra of [d(T*GGGT)](4) were characteristic of parallel-stranded, tetramolecular quadruplex. CD and DSC melting experiments reveal that [d(T*GGGT)](4) is less stable that unmodified quadruplex. Molecular models suggest possible explanation for the observed behaviour. PMID- 12568921 TI - Mesostructure of fibrillar bovine serum albumin gels. AB - The mesostructure of bovine serum albumin (BSA) at low pH was investigated. Rheological measurements were performed to determine the critical percolation concentration (c(p)). A decreasing c(p) with increasing ionic strength was found. Fibrils with a contour length of about 100-300 nm were found using transmission electron microscopy. The measured conversion of monomers into fibrils was independent of ionic strength (0.20-0.30 M). Dilution of BSA samples showed that the aggregation process is reversible and that there exists a critical concentration for the self-assembly of BSA. We explain the decreasing c(p) with increasing ionic strength in terms of an adjusted random contact model. PMID- 12568922 TI - Thermoreversible gelation in aqueous binary solvents of chemically modified agarose. AB - The thermoreversible gelation of chemically modified agarose has been studied in aqueous binary solvents (dimethyl sulfoxide and a series of formamide) by differential calorimetry, mechanical testing, and small-angle neutron scattering. The temperature-composition phase diagrams have been established. It is concluded that gelation is promoted by the formation of ternary complexes modified agarose/water/cosolvent, wherein the cosolvent mediates the interaction between chains through the formation of electrostatic interactions. PMID- 12568923 TI - Salt influence on glutathione--Schistosoma japonicum glutathione S-transferase binding. AB - There has been some speculation about the salt independence of Schistosoma japonicum glutathione S-transferase (Sj26GST, EC. 2.5.1.18), but this aspect has not been carefully studied before. To establish the basis for a further development of this dependence, we have performed a methodical study of the influence of some important ions and their concentration on the binding properties of glutathione to Sj26GST by means of isothermal calorimetry and fluorescence quenching. Salts like NaCl, Na(2)SO(4) and MgSO(4) do not change practically the affinity of the protein for its substrate, whilst MgCl(2) has the effect of decreasing the affinity as its concentration rises. However, the enthalpy change is not affected by all the salts studied, and so, the entropy change is the causal factor in dropping the affinity. We also looked at the conformational stability of the protein under different conditions to check the structural changes they provide, and found that the unfolding parameters are practically not affected by the salt concentration. We discuss the results in terms of the chaotropic nature of the ions implied. PMID- 12568924 TI - The effect of copper/zinc replacement on the folding free energy of wild type and Cys3Ala/Cys26Ala azurin. AB - The effect of copper/zinc metal ion replacement on the folding free energy of wild type (w.t.) and disulfide bridge depleted (C3A/C26A) azurin has been investigated by differential scanning calorimetry (DSC) and fluorescence techniques. The denaturation experiments have shown that, in both cases, the thermal transitions of the zinc derivative of azurins can be depicted in terms of the classical Lumry-Eyring model, N if U-->F, thus resembling the unfolding path of the two copper proteins. The thermally induced transition of Zn azurin, monitored by fluorescence occurs at lower temperature than the DSC scans indicating that a local conformational rearrangement of the Trp microenvironment, takes place before protein denaturation. For Zn C3A/C26A azurin, the two techniques reveal the same transition temperature. Comparison of the thermodynamic data shows that the presence of Zn in the active site stabilises the three-dimensional structure of azurin only when the disulfide bridge is present. Compared to the copper form of the protein, the unfolding temperature of Zn azurin has increased by 4 degrees C, while the unfolding free energy, deltaG, is 31 kJ/mol higher. Both enthalpic and entropic factors contribute to the observed DeltaG increase. However, the copper/zinc replacement has no effect on the unfolding free energy of C3A/C26A azurin. Taking Cu azurin w.t. as the reference state, for both Cu and Zn C3A/C26A azurin the unfolding free energy is decreased by about 28 kJ/mol, indicating that metal substitution is not able to compensate the destabilising effect induced by the disulfide bridge depletion. It is noteworthy that the thermal denaturation of the Zn derivative, which thermodynamically is the most stable form of azurin, is also characterized by the highest value of the activation energy, E(a), as derived from the kinetic stability analysis. PMID- 12568925 TI - Antimicrobial activity of acidic xylo-oligosaccharides produced by family 10 and 11 endoxylanases. AB - Acidic oligosaccharides were obtained from birchwood xylan by treatment with a Thermoascus aurantiacus family 10 and a Sporotrichum thermophile family 11 endoxylanases. The main difference between the products liberated by xylanases of family 10 and 11 concerned the length of the products containing 4-O-methyl-D glucuronic acid. The xylanase from T. aurantiacus liberate from glucuronoxylan an aldotetrauronic acid as the shortest acidic fragment in contrast with the enzyme from S. thermophile, which liberated an aldopentauronic acid. Acidic xylooligosaccharides were separated from the hydrolysate by anion-exchange and size-exclusion chromatography (SEC) and the primary structure was determined by 13C NMR spectroscopy. The acidic xylo-oligosaccharides were tested against three Gram-positive and three Gram-negative aerobically grown bacteria, as well as against Helicobacter pylori. Aldopentauronic acid was proved more active against the Gram-positive bacteria and against H. pylori. PMID- 12568926 TI - Cation effects on sol-gel and gel-sol phase transitions of kappa-carrageenan water system. AB - Sol-gel and gel-sol phase transitions of kappa-carrageenan in pure water and in KCl solution were studied using photon transmission technique. Photon transmission intensity, I(tr), was monitored against temperature to determine the sol-gel and gel-sol temperatures (T(sg) and T(gs)) and activation energies (deltaH(sg) and deltaH(gs)). It was observed that T(gs) was notably higher than T(sg) due to the hysteresis on the phase transition loops. T(gs) and deltaH(gs) values were also higher for gels containing KCl than for those without KCl. The increase in carrageenan content caused an increase in both critical temperatures and activation energies for the gels prepared in pure water and in KCl solution. Increases in the KCl/carrageenan ratio, raised both T(gs) and T(sg). Similarly deltaH(sg) was elevated by the increase in cation content of the gel. These results were interpreted as the formation of stronger gels in the presence of KCl in water. PMID- 12568927 TI - Behavior of various mammalian albumins towards bilirubin binding and photochemical properties of different bilirubin-albumin complexes. AB - Bilirubin (BR) binding properties of serum albumins from different mammalian species viz. human (HSA), equine (ESA), dog (DSA) and guinea pig (GPSA) were studied by absorption, fluorescence and CD spectroscopy. Whereas, a complex of BR with ESA produced maximum change, GPSA-BR complex showed weaker interaction as reflected from absorption and fluorescence spectroscopic data. Conformational analysis of these albumins by near- and far-UV CD spectra suggested similar structural characteristics (both secondary and tertiary structures) for ESA and HSA, whereas, DSA and GPSA had lower amounts of secondary and tertiary structures being minimum for GPSA. Photoirradiation results of BR-albumin complexes showed GPSA-bound BR more labile compared with other complexes, whereas, BR-ESA complex was found to be more stable against photoinduced chemical changes. Taken together, all these results suggest that chiroptical properties/stability of albumin bound BR varies with albumin species. PMID- 12568928 TI - Molecular characterization and properties of (R)-specific enoyl-CoA hydratases from Pseudomonas aeruginosa: metabolic tools for synthesis of polyhydroxyalkanoates via fatty acid beta-oxidation. AB - The use of (R)-specific enoyl-coenzyme A (CoA) hydratase (PhaJ) provides a powerful tool for polyhydroxyalkanoate (PHA) synthesis from fatty acids or plant oils in recombinant bacteria. PhaJ provides monomer units for PHA synthesis from the fatty acid ss-oxidation cycle. Previously, two phaJ genes (phaJ1(Pa) and phaJ2(Pa)) were identified in Pseudomonas aeruginosa. This report identifies two new phaJ genes (phaJ3(Pa) and phaJ4(Pa)) in P. aeruginosa through a genomic database search. The abilities of the four PhaJ(Pa) proteins and the (R)-3 hydroxyacyl-acyl carrier protein [(R)-3HA-ACP] dehydrases, FabA(Pa) and FabZ(Pa), to supply monomers from enoyl-CoA substrates for PHA synthesis were determined. The presence of either PhaJ1(Pa) or PhaJ4(Pa) in recombinant Escherichia coli led to the high levels of PHA accumulation (as high as 36-41 wt.% in dry cells) consisting of mainly short- (C4-C6) and medium-chain-length (C6-C10) 3HA units, respectively. Furthermore, detailed characterizations of PhaJ1(Pa) and PhaJ4(Pa) were performed using purified samples. Kinetic analysis revealed that only PhaJ4(Pa) exhibits almost constant maximum reaction rates (V(max)) irrespective of the chain length of the substrates. The assay for stereospecific hydration revealed that, unlike PhaJ1(Pa), PhaJ4(Pa) has relatively low (R)-specificity. These hydratases may be very useful as monomer-suppliers for the synthesis of designed PHAs in recombinant bacteria. PMID- 12568929 TI - Inhibition of cryogelation by the novel synthetic peptide (Gly-Arg-Lys-Lys-Thr): recognition site of extra domain A containing fibronectin for heparin in cryogelation. AB - Cryogel is a physical gel formed by the heterophilic aggregation of extra domain A (EDA) containing fibronectin [EDA(+)FN], plasma fibronectin (pFN), fibrinogen (Fbg) and heparin (Hep) in the blood of rheumatoid arthritis (RA) patients. In cryogelation EDA(+)FN cross-links to form an interaggregate of cryogel with Hep. In the present study, we determined the recognition structure of Hep for EDA(+)FN by using oligo- and desulfonated-Hep. The affinity constant (KA) (1.2 x 10(8) per M) of oligo-Hep for EDA(+)FN did not change with a decrease in number-average molecular weight (4.9 x 10(4)-->6.0 x 10(3)). The KA-value of desulfonated-Hep for EDA(+)FN decreased from 3.2 x 10(8) to 1.0 x 10(7) per M with a decrease in the sulfonation ratio (7.0-->4.3%). We also determined the recognition structure of EDA(+)FN for Hep by an inhibition experiment on the heparin binding domain II (HepII) in EDA(+)FN with the synthetic peptides, Arg-Arg-Ala-Arg (RRAR), Asp-Gln Ala-Arg (DNAR), Ile-Lys-Tyr-Glu-Lys (IKYEK), and Gly-Arg-Lys-Lys-Try (GRKKT). The GRKKT sequence clearly inhibited bonding between EDA(+)FN and Heps containing oligo- and desulfonated-Hep. The amount of cryogel formed in the RA-patient model plasma corresponded to the EDA(+)FN concentration in cryogel (36.7%) normalized by the EDA(+)FN concentration in plasma. When GRKKT was added to plasma, the EDA(+)FN concentration fell to 10.5%. These results demonstrated that inhibition of cryogelation in plasma could progress to a novel treatment for RA. PMID- 12568930 TI - Interaction of a low mobility group protein, LMG160, with deoxyribonucleic acid. AB - A fraction of low mobility group (LMG) nonhistone protein designated LMG(160) was isolated from rat liver chromatin by preparative gel electrophoresis and its interaction with DNA was studied using thermal denaturation and DNA-cellulose affinity chromatography techniques. The results showed that LMG(160) with an isoelecteric point of 5-5.5 was bound to DNA and decreased its melting temperature. Increasing ionic strengths decreased this effect. DNA-cellulose affinity chromatography showed the affinity of LMG(160) to double stranded DNA was higher than that to single stranded DNA, since it required 0.6 M NaCl for elution. The results suggest that LMG(160) protein preferentially binds to double stranded DNA destabilizes it and the binding is electrostatic. PMID- 12568931 TI - Mg2+ ion effect on conformational equilibrium of poly A . 2 poly U and poly A poly U in aqueous solutions. AB - Differential UV spectroscopy and thermal denaturation were used to study the Mg(2+) ion effect on the conformational equilibrium in poly A.2 poly U (A2U) and poly A . poly U (AU) solutions at low (0.01 M Na(+)) and high (0.1 M Na(+)) ionic strengths. Four complete phase diagrams were obtained for Mg(2+)-polynucleotide complexes in ranges of temperatures 20-96 degrees C and concentrations (10(-5) 10(-2)) M Mg(2+). Three of them have a 'critical' point at which the type of the conformational transition changes. The value of the 'critical' concentration ([Mg(t)(2+)](cr)=(4.5+/-1.0) x 10(-5) M) is nearly independent of the initial conformation of polynucleotides (AU, A2U) and of Na(+) contents in the solution. Such a value is observed for Ni(2+) ions too. The phase diagram of the (A2U+Mg(2+)) complex with 0.01 M Na(+) has no 'critical' point: temperatures of (3-->2) and (2-->1) transitions increase in the whole Mg(2+) range. In (AU+Mg(2+)) phase diagram at 0.01 M Na(+) the temperature interval in which triple helices are formed and destroyed is several times larger than at 0.1 M Na(+). Using the ligand theory, a qualitative thermodynamic analysis of the phase diagrams was performed. PMID- 12568932 TI - Application of CP-MAS and liquid-like solid-state NMR experiments for the study of the ripening-associated cell wall changes in tomato. AB - 13C and 1H NMR spectra of an ethanol insoluble material (EIM) prepared from the pericarp of mature-green (MG) and red-ripe (RR) tomato fruits were acquired in 'liquid-like' and cross-polarisation with dipolar decoupling and magic angle spinning (CPMAS) conditions using the same triple resonance probe. Such a strategy allowed acquisitions of various NMR experiments aimed at detecting compositional differences as well as distinguishing differences in molecular mobility for various constituent polysaccharides related with the two ripening stages. Increase of the proton dipolar decoupling power levels from 3 to 50-55 kHz during single pulse 13C acquisition, led to more intense signals for pectic and hemicellulosic polysaccharides. This behaviour was interpreted as reflecting motional restrictions of these polysaccharides inside the porous cell wall network. Measurements of the proton rotating frame relaxation times T(1rho) in the 'liquid-like' conditions and of the proton transverse relaxation times T(2) from CPMAS spectra, revealed changes in mobilities for some pectic polysaccharides in relation with ripening, particularly for the H1 and H5 protons of alpha-1,5 arabinan (Ara) side chains of rhamnogalacturonans. These data are discussed in relation with known pectic modifications occurring during ripening and associated with the tomato fruit softening. PMID- 12568933 TI - Ferricytochrome c encapsulated in silica hydrogels: correlation between active site dynamics and solvent structure. AB - Ferricytochrome c encapsulated in silica hydrogels has been prepared by the sol gel technique following, with some modifications, the procedure originally developed by Zink et al. A suitable preparation of hydrogels enables to have both 'wet' and 'dry' samples. Wet samples have a high water content: as the temperature is lowered below approximately 260 K water freezes and the samples crack. On the contrary, dry samples have a low water content (hydration h approximately 0.35): in these conditions water does not freeze even at cryogenic temperatures and the samples remain transparent and non-cracking. The dynamics of ferricytochrome c and its dependence on the surrounding medium have been studied by optical absorption spectroscopy in the temperature range 10-300 K. At each temperature, spectra were collected both in the Soret region and in the near infrared at approximately 1.45 microm (the water overtone band); this enables to probe the local dynamics of the protein active site as well as the 'structure' of water molecules present in the sample. The data show that sol-gel encapsulation 'per se' does not alter the protein active site dynamics, but rather introduces an increased local heterogeneity. At difference, we find a correlation between active site dynamics and water structure: in the wet hydrogel, freezing of water quenches the ensemble of soft modes linearly coupled to the Soret transition; while, in the dry hydrogel, water does not freeze, and an active site dynamic behavior-similar to the non-freezing water/glycerol solution-is observed. PMID- 12568934 TI - YY1-DNA interaction results in a significant change of electronic context as measured by capacitance. AB - The detailed mechanism behind the processes of DNA-dependent RNA transcription initiation is largely unknown. When transcription initiation factors bind DNA, a significant change in the electrostatic state of the complex can result. Using electrical capacitance measurements of solutions of the YY1 zinc finger transcription initiation factor and the adeno-associated viral P5 promoter DNA, we observed a specific dielectric change when a protein-DNA complex was formed. We propose that complexation results in electrostatic changes that may trigger the markedly different electrical behavior, and offer a possible explanation for our results. PMID- 12568935 TI - Mechanistic equations for membrane substance transport and their identity with Kedem-Katchalsky equations. AB - Since the physical interpretation of practical Kedem-Katchalsky (KK) equations is not clear, we consider an alternative, mechanistic approach to membrane transport generated by osmotic and hydraulic pressure. We study a porous membrane with randomly distributed pore sizes (radii). We postulate that reflection coefficient (sigma(p)) of a single pore may equal 1 or 0. From this postulate we derive new (mechanistic) transport equations. Their advantage is in clear physical interpretation and since we show they are equivalent to the KK equations, the interpretation of the latter became clearer as well. Hence the equations allow clearer and more detailed interpretation of results concerning membrane substances transport. PMID- 12568936 TI - Assembly of EcoKI DNA methyltransferase requires the C-terminal region of the HsdM modification subunit. AB - The methyltransferase component of type I DNA restriction and modification systems comprises three subunits, one DNA sequence specificity subunit and two DNA modification subunits. Limited proteolysis of the EcoKI methyltransferase shows that a 55-kDa N-terminal fragment of the 59-kDa modification subunit is resistant to degradation. We have purified this fragment and determined by mass spectrometry that proteolysis removes 43 or 44 amino acids from the C-terminus. The fragment fails to interact with the other subunits even though it still possesses secondary and tertiary structure and the ability to bind the S adenosylmethionine cofactor. We conclude that the C-terminal region of the modification subunit of EcoKI is essential for the assembly of the EcoKI methyltransferase. PMID- 12568937 TI - Molecular dynamics investigation of an antibody binding site by the fluorescence photochrome method. AB - A combined fluorescence-photochrome approach was used for investigation of the molecular dynamics antiDNP antibody binding site and its cavity. A 4-(N-2,4 dinitrophenylamino)-4'-(N,N'-dimethylamino)stilbene (StDNP) fluorescence DNP analog was incorporated into the antibody binding site. This was followed by measurements of fluorescence and photochrome parameters such as the StDNP excitation and emission spectra, fluorescence lifetime, steady-state and time resolved fluorescence polarization, kinetics of trans-cis and cis-trans photoisomerization, and fluorescence quenching by nitroxide radicals freely diffused in solution. In parallel, computational modeling studies on the location and dynamics of DNP/TEMPO spin-label (NslDNP) and StDNP guests within a model of the binding site were performed. When all the experimental evidence is considered (including data from the antibody X-ray study), one can conclude that wobbling of the Trp 91 L/Trp 96 H binding-site.bound-hapten moiety (StDNP), can be responsible for the label's nanosecond dynamics monitored by fluorescence polarization techniques. A similar conclusion may be reached as a result of data analysis on NslDNP mobility within the antibody binding site. The mobility of Trp 91 L and Trp 96 H moieties provides the induced fit needed for effective stacking and release of the DNP epitope. Analysis of the above-mentioned data allows one to explore the mechanism of the probe's movement within the binding site and enables one to discuss the local dynamics of the binding site region. The combined fluorescence-photochrome approach can be used for investigation of local medium molecular dynamics in the immediate vicinity of specific sites of proteins and nucleic acids, as well as for other biologically important structures and synthetic analogues. PMID- 12568938 TI - Electrochemical investigations of cholesterol enriched glassy carbon supported thin lipid films. AB - The formation and study of stable cholesterol enriched thin lipid layers onto the surface of glassy carbon electrode is reported in this work. The method of formation relies on additional thinning of wetting films by electrostriction. Electrochemical techniques based on the concepts of impedance and voltammetry are used to explore the films' features. The impedance data reveal a substantial change of relaxation characteristics of the modified films. In this respect, opportunities for the evaluation of the films' stage based on the approximation with 'constant phase angle element' are discussed. The possible final structure of the films, as well as, their relevance for development of sensor elements are briefly viewed. PMID- 12568939 TI - Enzyme catalysis in microgravity: steady-state kinetic analysis of the isocitrate lyase reaction. AB - Two decades of research in microgravity have shown that certain biochemical processes can be altered by weightlessness. Approximately 10 years ago, our team, supported by the European Space Agency (ESA) and the Agenzia Spaziale Italiana, started the Effect of Microgravity on Enzyme Catalysis project to test the possibility that the microgravity effect observed at cellular level could be mediated by enzyme reactions. An experiment to study the cleavage reaction catalyzed by isocitrate lyase was flown on the sounding rocket MASER 7, and we found that the kinetic parameters were not altered by microgravity. During the 28th ESA parabolic flight campaign, we had the opportunity to replicate the MASER 7 experiment and to perform a complete steady-state analysis of the isocitrate lyase reaction. This study showed that both in microgravity and in standard g controls the enzyme reaction obeyed the same kinetic mechanism and none of the kinetic parameters, nor the equilibrium constant of the overall reaction were altered. Our results contrast with those of a similar experiment, which was performed during the same parabolic flight campaign, and showed that microgravity increased the affinity of lipoxygenase-1 for linoleic acid. The hypotheses suggested to explain this change effect of the latter were here tested by computer simulation, and appeared to be inconsistent with the experimental outcome. PMID- 12568940 TI - Kinetic isotope effects of proton transfer in aqueous and methanol containing solutions, and in gramicidin A channels. AB - The electrochemical conductivities of HCL and DCI were measured in: H(2)O and D(2)O; in methanol and fully deuterated methanol; and in water-methanol solutions. The single channel conductances to H(+) (g(H)) and D(+) (g(D)) in various gramicidin A (gA) ion channels incorporated in glycerylmonooleate planar bilayers were also measured. Kinetic isotope effects (KIE) were estimated from the ratio of conductivity measurements. In 1 and 5 M HCl aqueous solutions and in 1 M HCl+3.7 M methanol, the KIE ( approximately 1.35) is not different from values previously determined in dilute acid solutions. This suggests that the mobility of protons in those solutions is largely determined by proton transfer. In 10 M HCl, however, where the mobility of protons is likely to be determined by hydrodynamic diffusion, the measured KIE is considerably larger (1.47). Possible causes for this effect are discussed. The KIE of proton conductivities in 5 and 50 mM HCl in methanol and d-methanol is approximately 1.15. This is considerably smaller than the ratio between conductivities of 5 mM KCl in methanol and d methanol (1.24). The KIE values (1.22-1.37) for g(H) in gA channels in 1 M HCl are significantly larger than for other monovalent cations and consistent with H(+) transfer. Methanol reduces g(H) in gA channels. The KIE of this effect is not different from the one measured in the absence of methanol. Possible mechanisms for the methanol-induced block of H(+) conductivities in solution and gA channels are discussed. PMID- 12568941 TI - Tropicamide effects on pupil size and pupillary light reflexes in Alzheimer's and Parkinson's disease. AB - Diagnostic tests for Alzheimer's disease (AD) involving tropicamide blockade of cholinergic oculomotor functions were examined in AD patients (n=15), Parkinson's disease (PD) patients (n=15), and non-clinical control (NC) participants (n=15). Pupillographic methods were used to measure pupil diameter and pupillary light reflexes after double-blind ocular administration of dilute tropicamide (0.01%) in one eye and saline in the other eye. Changes in pupil size were measured in bright background light and near-darkness. Tropicamide increased pupil diameter to a similar extent in all three groups in light and darkness. Tropicamide also reduced the amplitude and latency of the pupillary light reflex to a similar extent for all three groups. Tropicamide pupillary response tests, therefore, were not sensitive or specific diagnostic tests for AD. Peak constriction amplitude of the pupillary light reflex was significantly reduced in both eyes in AD and PD groups relative to non-clinical controls, but AD and PD groups did not differ significantly. The pupillary light reflex test, therefore, was sensitive to AD, but lacked adequate specificity. Finally, peak constriction amplitude correlated significantly with dementia severity and donepezil treatment may have partially normalized pupillary light reflex abnormalities in AD patients. The pupillary light reflex test, therefore, may index central cholinergic dysfunction associated with disease progression and improvement in cholinergic function associated with pharmacologic treatment response in AD. PMID- 12568942 TI - Effects of effort and distress coping processes on psychophysiological and psychological stress responses. AB - The purpose of this study was to investigate the effects of coping processes on psychophysiological and psychological responses in stressful settings. In particular, we focused on the effects of a combination of active and avoidant coping processes. Subjects were 40 healthy undergraduate male students (mean=19.80, S.D.=0.97) who were randomly divided into four groups: (a) an effort coping group in which a subject faced a controllable stressor mobilizing an effortful and active coping behavior for a reward; (b) a distress coping group in which a subject faced a distressful stressor mobilizing an avoidant coping behavior for threat of punishment; (c) an effort-distress coping group in which a subject faced an ambivalent stressor mobilizing active coping behavior for a reward and avoidant coping behavior for threat of punishment; and (d) a control group. Initially, the effects of effort coping, distress coping, and effort distress coping on psychophysiological and psychological responses were investigated. It was found that effort coping and effort-distress coping intensified cardiovascular responses, particularly blood pressure, and that distress coping and effort-distress coping intensified skin conductance level (SCL). Secondarily, the relationships between effort coping process, distress coping process, psychophysiological responses, and psychological responses were investigated. As a result of cluster analysis, the changes of heart rate and blood pressure were correlated to the change of the effort score, and the changes of SCL and psychological responses were correlated to the change of the distress score. These findings suggest that active coping processes and avoidant coping processes independently affect different response systems. PMID- 12568943 TI - Effects of stimulant medications on the EEG of children with Attention Deficit/Hyperactivity Disorder Predominantly Inattentive type. AB - Stimulant medications are the most commonly-used treatments for Attention Deficit/Hyperactivity Disorder (ADHD) in North America and Australia, although it is still not entirely known how these medications work. This study investigated the effects of stimulant medications on the EEG of children with the Inattentive type of ADHD. An initial EEG was recorded during an eyes-closed resting condition and Fourier transformed to provide absolute and relative power estimates for the delta, theta, alpha and beta bands. Theta/alpha and theta/beta ratios were also calculated. Subjects were placed on a 6-month trial of a stimulant and a second EEG was recorded at the end of the trial. Subjects were included in this study only if they showed a good clinical response during the trial. The unmedicated ADHD group had significantly greater absolute and relative theta, less relative alpha, and higher theta/alpha and theta/beta ratios than the control group. The stimulant medications resulted in a normalisation of the EEG, with changes in the theta, alpha and beta bands being most evident. These results suggest that stimulants act to increase cortical arousal in children with ADHD, normalising their EEG. PMID- 12568944 TI - Facilitation of heartbeat self-detection in a choice task. AB - Brener and Kluvitse (Psychophysiology 25 (1988) 554) developed a heartbeat self detection task in which subjects are required to judge the simultaneity of heartbeat sensations and tones presented at six intervals following the onset of each ventricular contraction (0, 100, 200, 300, 400 or 500 ms). They found that most of their participants considered as simultaneous in the intervals of 100, 200 and 300 ms, in comparison to those of 0, 400 and 500 ms. In the research reported here, an adaptation of this task was administered to 32 subjects (16 men, 16 women) to see whether these interval choices would be replicated using a narrower range of values, and if task performance would be affected by the reduction in number of trials from 30 to 20. The interval values of the external stimuli with reference to the R-wave were 150, 200, 250, 300 and 350 ms. The results indicate that there was a clear choice for the intervals of 150 and 200 ms in comparison to the rest and that the performance of participants decreases after 20 trials. PMID- 12568945 TI - Pain in the trigeminal system: irritation of the nasal mucosa using short- and long-lasting stimuli. AB - The paper describes methods which allow intranasal irritation using short- and long-lasting painful stimuli in humans. Short-lasting pain is induced by gaseous CO(2), while long-lasting pain is induced by a stream of dry air. Both models have been explored regarding their major determinants, e.g. stimulus duration, stimulus intensity, or repeated stimulation. Short-lasting, non-inflammatory pain stimuli seem to provide specific indicators of A(delta)-fiber function, while responses to long-lasting, inflammatory pain appear to be indicative of C-fiber function. Responses to both types of painful stimuli are modulated by analgesic drugs. As these well-investigated models allow the detailed and precise analysis of modulatory effects on intranasal nociception, they appear to be suited for the investigation of subtle changes of intranasal irritation, e.g. induced by environmental agents. PMID- 12568946 TI - Relative effects of harassment, frustration, and task characteristics on cardiovascular reactivity. AB - Effects of anger induction procedures such as frustration and harassment on cardiovascular reactivity have been demonstrated in a wide range of experimental situations. Similarly, heightened cardiovascular reactivity has been associated with a diverse range of tasks involving active coping, competition and interpersonal interaction. The present study sought to directly compare the relative effects of these two important ways of inducing cardiovascular changes. One hundred and five university students performed two tasks that differed in the degree of active coping and interpersonal competition: a competitive psychomotor task and a problem-solving task. States of anger were induced during both tasks by means of harassment, frustration or frustration+harassment. Task-related changes in heart rate, systolic blood pressure, diastolic blood pressure, pulse volume amplitude and respiratory sinus arrhythmia amplitude were monitored. The competitive psychomotor task produced greater cardiovascular reactivity than did the problem-solving task. Harassment and frustration+harassment provoked more cardiovascular reactivity than did frustration alone. However, harassment and frustration+harassment had the greatest cardiovascular effects in the competitive task, whereas frustration had the greatest cardiovascular effects in the problem solving task. In this sense, the increases on cardiovascular reactivity seem to depend on the interaction between anger induction procedures and the context in which anger is provoked. PMID- 12568947 TI - The relationship between reaction time, error rate and anterior cingulate cortex activity. AB - Current concepts of cognitive control suggest that the anterior cingulate cortex (ACC) is involved in performance monitoring. This idea is supported by the finding that increased ACC activity is found in situations in which errors are likely to occur, even if none are actually made. In addition, recent results suggest that increased ACC activity is negatively correlated with reaction time. We have now compared the error rates and the ACC activity of healthy subjects with short (n=19) vs. long reaction times (n=17) in an auditory choice reaction paradigm and analysed the current density differences in the ACC in the time range of the N1 component with low resolution electromagnetic tomography. Subjects with short reaction times showed significantly more ACC activation (Brodmann Area 24) and an increased error rate. This finding suggests that increased ACC activity is associated with a gain in reaction speed at the expense of correctness and is discussed in the context of current concepts about the role of the ACC in cognitive functions. PMID- 12568948 TI - Four-year study of cobalt and chromium blood levels in patients managed with two different metal-on-metal total hip replacements. AB - BACKGROUND: Metal-on-metal total hip prostheses will produce a certain amount of wear debris. This results in increased whole-blood metal levels, which may cause adverse effects. It is not known to what extent the problem has been overcome by advances in alloy technology. METHODS: In 259 patients who with total hip replacement, blood cobalt and chromium concentrations were measured with atomic absorption spectrophotometry over a period of four years after arthroplasty. Of the patients enrolled in the study, 131 had been managed with a METASUL cobalt chromium alloy metal-on-metal bearing combination, while 128 had been given a SIKOMET-SM21 cobalt-chromium alloy metal-on-metal combination. The control group consisted of 31 age- and gender-matched subjects. RESULTS: Compared with the controls, all the patients had higher cobalt and chromium levels. Cobalt concentrations were up to 50 times higher, while chromium concentrations were up to 100 times higher. CONCLUSIONS: Both systems showed evidence, in the whole blood samples, of wear debris production by the implants. Therefore, patients managed with metal-on-metal bearing combinations should be carefully monitored in order to ensure that any local or systemic complications are detected early on. PMID- 12568949 TI - PMMA particles and pressure--a study of the osteolytic properties of two agents proposed to cause prosthetic loosening. AB - Amongst the wear debris particles implicated in the particle hypothesis for prosthetic loosening are polymethylmethacrylate (PMMA), and particularly PMMA with barium sulphate contrast agent. Another suggested cause for loosening is hydrostatic pressure. PMMA particles were combined with hydrostatic pressure in a study to investigate whether there could be a synergistic or additive effect between these two factors. Titanium plates were fastened onto tibiae of 59 rats. After osseointegration, PMMA particles with barium sulphate were administered to the bone-implant interface. Further, PMMA particles were introduced into a previously published model for hydrostatic pressure induced osteolysis. There was measurable resorption in response to the PMMA particles but no additive or synergistic effect from introducing particles to the pressure model, and the effect of pressure was far greater than that of particles. These results suggest that, whereas particles can be shown to elicit an osteolytic response, the much less studied osteolytic effects of pressure could be far more important. PMID- 12568951 TI - Autologous osteoblasts enhance osseointegration of porous titanium implants. AB - The goal of this study was to assess the osseointegration of porous titanium implants by means of coating with autologous osteoblasts. Titanium implants (8 x 5 x 4 mm) having drill channels with diameters of 400, 500, and 600 microm were coated with autologous osteoblasts obtained from spongiosa chips. The implants were inserted into the distal femora of 17 adult Chinchilla Bastard rabbits (group I). Uncoated implants were inserted as controls in the contralateral femur (group II). The animals were sacrificed after 5, 11, and 42 days. Intravital fluorochrome labeling and microradiography were used for the assessment of bone ingrowth into the titanium channels. In both groups, no bone tissue was formed in the channels up to day 5. On day 11, group I exhibited significantly more (p<0.05) bone tissue (19.8+/-14.0% vs. 5.8+/-9.1%) with greater bone-implant contact (13.3+/-15.1% vs. 5.7+/-5.3%, p<0.05) at the channel mouths than group II. Bone tissue was formed mainly between day 15 and 30 in group I, in group II between day 25 and 40. Six weeks after implantation, bone tissue filled on an average 68.8+/-15.1% of the mouths of the drill channels in implants in group I, the filling for group II was 49.8+/-18.1% (p<0.05). The average bone-implant contact at the channel mouths after six weeks was 56.5+/-13.5% in group I, 40.2+/ 21.9% in group II (p<0.05). 600-microm channels showed at this time point the best osseous integration (p<0.05). Coating with autologous osteoblasts accelerates and enhances the osseointegration of titanium implants and could be a successful biotechnology for future clinical applications. PMID- 12568950 TI - RANKL is an essential cytokine mediator of polymethylmethacrylate particle induced osteoclastogenesis. AB - RANKL is a TNF superfamily member and an essential cytokine mediator of developmental osteoclastogenesis. We examined the role of RANKL in PMMA particle induced osteoclastogenesis in vitro. In murine whole bone marrow cultures, PMMA particles stimulate a 2.5 fold increase in secreted RANKL, a 5-8 fold increase in osteoclast number and induce the formation of giant multinuclear osteoclasts. RANKL and TNF, potential cytokine mediators of PMMA, had similar osteoclastogenic effects. The RANKL inhibitor OPG was utilized to define the role of RANKL in mediating the PMMA response and was found to inhibit basal and PMMA particle induced osteoclastogenesis. Additionally, particles stimulate osteoclast formation in RANKL-primed osteoclast precursor cells (devoid of supporting stromal cells) while RANKL untreated osteoclast precursors demonstrate no osteoclastogenic response to particles. Since TNF can potentiate RANKL action and is thought to mediate implant osteolysis we analyzed TNF(-/-) whole bone marrow cultures to elucidate the role of this cytokine. In TNF(-/-) cultures basal osteoclastogenesis remains intact, yet the PMMA effect is blunted. Finally, we show that PMMA, RANKL and TNF all activate the NF-kB and c-jun/AP-1 signaling pathways which are both fundamental to osteoclast formation and are potential sites of signal convergence in RANKL-mediated particle osteoclastogenesis. PMID- 12568953 TI - The relationship between knee strength and functional stability before and after anterior cruciate ligament reconstruction. AB - Functional stability of the knee is dependent on an intact ligamentous system and the timely and efficient contraction of supporting musculature. The aim of this study was to assess the relationship between muscle strength and functional stability in 31 patients pre- and post-operatively, following a unilateral anterior cruciate ligament rupture. All subjects underwent reconstructive surgery using semitendonosis and gracilis tendons. Isokinetic strength assessment of quadriceps and hamstring muscles was performed at a rate of movement of 60 degrees /s and 120 degrees /s. Functional stability was determined by performance during five functional stability tests that included the shuttle run, side step, carioca, single and triple hop tests. Pearson's correlation coefficient statistics were applied to pre-operative and post-operative data respectively. These analyses demonstrated a significant positive correlation between quadriceps strength indices at both testing speeds and the two hop tests pre-operatively (p's<0.007) and between quadriceps strength indices at both speeds and all five functional tests post-operatively (p's<0.01). Assessed using Steiger's formula, there was a significant increase in the correlation between quadriceps strength indices and three functional tests post-operatively compared to pre-operatively (p<0.05). No significant correlation between hamstring strength indices and functional scores existed pre- or post-operatively. This study has shown a significant correlation exists between quadriceps strength indices and functional stability both before and after surgery, this relationship does not reach significance between hamstring strength indices and functional stability. PMID- 12568952 TI - A comparison of joint stability between anterior cruciate intact and deficient knees: a new canine model of anterior cruciate ligament disruption. AB - Transection of the canine anterior cruciate ligament (ACL) is a well-established osteoarthritis (OA) model. This study evaluated a new method of canine ACL disruption as well as canine knee joint laxity and joint capsule (JC) contribution to joint stability at two time points (16 and 26 weeks) after ACL disruption (n=5/time interval). Ten crossbreed hounds were evaluated with force plate gait analysis and radiographs at intervals up to 34 weeks after monopolar radiofrequency energy (MRFE) treatment of one randomly selected ACL. Each contralateral ACL was sham treated. The MRFE treated ACLs ruptured approximately eight weeks (mean 52.5 days, SEM+/-1.0, range 48-56 days) after treatment. Gait analysis and radiographic changes were consistent with established canine ACL transection models of OA. Anterior-posterior (AP) translation and medial-lateral (ML) rotation were measured in each knee at 30 degrees, 60 degrees, and 90 degrees of flexion with and then without JC with loads of 40 N in AP translation and 4 Nm in ML rotation. A statistically significant interaction in AP translation included JC by cruciate (P=0.02), and there was a trend for a cruciate by time (P=0.07) interaction. Significant interactions in ML rotational testing included the presence of joint capsule (P=0.0001) and angle by cruciate (P=0.0012). This study describes a model in which canine ACLs predictably rupture approximately eight weeks after arthroscopic surgery and details the contribution of JC to canine knee stability in both ACL intact and deficient knees. The model presented here avoids the introduction of potential surgical variables at the time of ACL rupture and may contribute to studies of OA pathogenesis and inhibition. This model may also be useful for insight into the pathologic changes that occur in the knee as the ACL undergoes degeneration prior to rupture. PMID- 12568954 TI - The effect of selected growth factors on human anterior cruciate ligament cell interactions with a three-dimensional collagen-GAG scaffold. AB - Our work focuses on development of a collagen-glycosamimoglycan (CG) scaffold to facilitate ligament healing in the gap between the ruptured ends of the human anterior cruciate ligament (ACL). In the present investigation, we evaluated the effects of selected growth factors on human ACL cell responses important in tissue regeneration, namely cell migration, proliferation, collagen production, and expression of alpha-smooth muscle actin (SMA). METHODS: Explants from six human ACLs were cultured on top of a CG scaffold. Culture conditions were with either 2% FBS (control), or 2% FBS supplemented with TGF-beta1, PDGF-AB, EGF, or FGF-2. Histologic cell distribution, total DNA content, proliferation rate, rate of collagen synthesis, scaffold diameter and percentage of SMA positive cells were determined at two, three and four weeks. RESULTS: The addition of TGF-beta1 to the culture medium resulted in increased cell number, increased collagen production and increased expression of SMA within the scaffold. Supplementation with PDGF-AB resulted in increased cell proliferation rates within the scaffold and increased collagen production. The addition of FGF-2 resulted in increased cell proliferation rates and slowed rates of scaffold shrinkage when compared with the control group. DISCUSSION: This work suggests that certain growth factors can alter the biologic functions of human ACL cells in a CG scaffold implanted as a bridge at the site of an ACL rupture. Based on these findings, the addition of selected growth factors to an implantable CG scaffold may facilitate ligament healing in the gap between the ruptured ends of the human ACL. PMID- 12568955 TI - The biomechanical response to doses of TGF-beta 2 in the healing rabbit medial collateral ligament. AB - Ligament injuries result in significant disability in over 100,000 patients each year. Despite current methods of treatment, 13% of patients with medial collateral ligament (MCL) injury develop early signs of arthritis, suggesting an incomplete return of knee stability. The principal hypothesis of this work was that the addition of TGF-beta 2 to the healing MCL would accelerate the development of scar strength and stiffness. Forty-four rabbits were divided evenly into four groups, with each group receiving either 0.1, 1 or 5 microg of TGF-beta 2 and the fourth group receiving 1 microg TGF-beta 2 and 1 microg of PDGF. Each rabbit underwent bilateral transection of the MCL, with one side having treatment with one of four doses of growth factor and the other side left untreated. All animals were sacrificed at 6 weeks and the structural properties of maximum load at failure, stiffness, and energy absorbed at failure measured. All treatment groups demonstrated an increase in scar mass, but no group had a significant increase in scar load at failure at 6 weeks. The addition of 0.1 microg TGF-beta 2 led to a significant increase in scar stiffness. The addition of PDGF had no significant effect on any of the parameters studied. This study suggests the mechanical stiffness, but not the load at failure, of ligament scar can be significantly altered by the administration of TGF-beta 2. PMID- 12568956 TI - Neurohistological examination of the inferior glenohumeral ligament of the shoulder. AB - The neural histology of the anterior band of the inferior glenohumeral ligament (IGHL) was studied in 11 fresh shoulder specimen using a special silver impregnation technique. Between the collagen fibers small myelinated and unmyelinated dendrites could be detected. The appearance of neurovascular structures in the adjacent synovial layer clearly exceeded the typical supply to soft tissues. Analysing about 11,000 sections Ruffini mechanoreceptors that are known to be slow adapting were found on the humeral insertion of the band. The sections containing these neural end organs were identified by means of transillumination and reflection-contrast microscopy and reconstructed using three-dimensional image processing. The presence of neural structures including Ruffini corpuscles in these most important passive stabilizers of the shoulder joint shows that these ligaments function also as an active safety device. There slow adaption is a prerequisite for muscular reflexes counteracting the tensile stresses to which the passive stabilizing structures of the shoulder are exposed. A disruption of the continuity of these structures by mechanical forces or surgery can reduce the biofeedback and proprioceptive quality and thus lead to a decrease of shoulder function and/or stability. These observations should be taken into account when planning surgical interventions involving the IGHL. Procedures like capsule shifts or plications may affect mechanoreceptor orientation and concentrations, thereby affecting the interaction between these structures and the synergistic muscles. When possible, these intervention should avoid receptor-dense regions while attempting to restore normal anatomical orientation and tissue tension. PMID- 12568957 TI - IL-1 beta induces COX2, MMP-1, -3 and -13, ADAMTS-4, IL-1 beta and IL-6 in human tendon cells. AB - Overuse injuries and trauma in tendon often involve acute or chronic pain and eventual matrix destruction. Anti-inflammatory drugs have been used as a treatment, however, the cellular and molecular mechanisms of the destructive processes in tendon are not clearly understood. It is thought that an inflammatory event may be involved as an initiating factor. Mediators of the inflammatory response include cytokines released from macrophages and monocytes. Interleukin-1 beta (IL-1 beta) is a candidate proinflammatory cytokine that is active in connective tissues such as bone and cartilage. We hypothesized that tendon cells would express receptors and respond to IL-1 beta in an initial "molecular inflammation" cascade, that is, connective tissue cell expression of cytokines that induce matrix destructive enzymes. This cascade results in expression of matrix metalloproteinases (MMPs) and aggrecanases that may lead to matrix destruction. Normal human tendon cells from six patients were isolated, grown to quiescence and treated with human recombinant IL-1 beta in serum-free medium for 16 h. Total RNA was isolated and mRNA expression assessed by semiquantitative RT-PCR. IL-1 beta (1 nM) induced mRNAs for cyclooxygenase 2 (COX2), MMP-1, -3, -13 and aggrecanase-1 as well as IL-1 beta and IL-6, whereas mRNAs for COX1 and MMP-2 were expressed constitutively. The IL-1 beta-treated tendon cells released prostaglandin E(2) (PGE(2)) in the medium, suggesting that the inducible COX2 catalyzed this synthesis. Induction of PGE(2) was detectable at 10 pM IL-1 beta. IL-1 beta also stimulated MMP-1 and -3 protein secretion. Induction of MMP-1 and -3 was detectable at 10 pM IL-1 beta. Post-injury or after some other inciting events, exogenous IL-1 beta released upon bleeding or as leakage of local capillaries may drive a proinflammatory response at the connective tissue cell level. The resulting induction of COX2, MMP-1 and -3 may underscore a potential for nonlymphocyte-mediated cytokine production of MMPs that causes matrix destruction and a loss of tendon biomechanical properties. Endogenous IL-1 beta might contribute to the process through a positive feedback loop by stimulating expression and accumulation of MMPs in the tendon matrix. PMID- 12568958 TI - Inhibition of tendon cell migration by dexamethasone is correlated with reduced alpha-smooth muscle actin gene expression: a potential mechanism of delayed tendon healing. AB - Local corticosteroid injection is commonly used to treat sports-related tendon injuries. However, isolated cases of tendon rupture following injection suggest that this treatment may impair the healing process. Tendon healing requires the migration of tendon cells to the repair site, followed by the proliferation and synthesis of the extracellular matrix. This study was designed to determine the effect of dexamethasone on the migration of tendon cells intrinsic to rat Achilles tendon at concentrations similar to those typically used for local injection treatment. Furthermore, the existence of a correlation between this effect and the expression of the contractile actin isoform, alpha-smooth muscle (SM) actin, which is associated with cell motility, was also examined. Using cultured tendon cells, migration was evaluated by counting the number of initial outgrowths from the tendon explants and by transwell filter migration assay. The distribution and assembly of alpha-SM actin were assessed by immunocytochemistry. The mRNA and protein expressions of alpha-SM actin were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. Dose dependent dexamethasone inhibition was demonstrated for both tendon cells outgrowth from the explants, ex vivo, and migration of tendon cells through the transwell filter, in vitro. Immunocytochemical staining revealed significant decreases in both the amount and assembly of alpha-SM actin in cells. Suppression of mRNA expression and protein level of alpha-SM actin was revealed from RT-PCR and Western blot analyses. In conclusion, dexamethasone inhibits tendon cell migration that is correlated with decreased gene expression of alpha-SM actin. PMID- 12568959 TI - Effects of a cultured autologous chondrocyte-seeded type II collagen scaffold on the healing of a chondral defect in a canine model. AB - Using a previously established canine model for repair of articular cartilage defects, this study evaluated the 15-week healing of chondral defects (i.e., to the tidemark) implanted with an autologous articular chondrocyte-seeded type II collagen scaffold that had been cultured in vitro for four weeks prior to implantation. The amount and composition of the reparative tissue were compared to results from our prior studies using the same animal model in which the following groups were analyzed: defects implanted with autologous chondrocyte seeded collagen scaffolds that had been cultured in vitro for approximately 12 h prior to implantation, defects implanted with autologous chondrocytes alone, and untreated defects. Chondrocytes, isolated from articular cartilage harvested from the left knee joint of six adult canines, were expanded in number in monolayer for three weeks, seeded into porous type II collagen scaffolds, cultured for an additional four weeks in vitro and then implanted into chondral defects in the trochlear groove of the right knee joints. The percentages of specific tissue types filling the defects were evaluated histomorphometrically and certain mechanical properties of the repair tissue were determined. The reparative tissue filled 88+/-6% (mean+/-SEM; range 70-100%) of the cross-sectional area of the original defect, with hyaline cartilage accounting for 42+/-10% (range 7-67%) of defect area. These values were greater than those reported previously for untreated defects and defects implanted with a type II collagen scaffold seeded with autologous chondrocytes within 12 h prior to implantation. Most striking, was the decreased amount of fibrous tissue filling the defects in the current study, 5+/-5% (range 0-26%) as compared to previous treatments. Despite this improvement, indentation testing of the repair tissue formed in this study revealed that the compressive stiffness of the repair tissue was well below (20 fold lower stiffness) that of native articular cartilage. PMID- 12568960 TI - Type II collagen modulates the composition of extracellular matrix synthesized by articular chondrocytes. AB - The articular cartilage extracellular matrix (ECM) interfaces with chondrocytes and influences many biological processes important to cartilage homeostasis and repair. The alginate bead culture system can be viewed as a model of cartilage repair in which the chondrocyte attempts to recreate the pericellular matrix while maintaining a differentiated phenotype. The purpose of this study was to evaluate the alteration in epitopes of proteoglycan and tenascin synthesized by chondrocytes in the presence of exogenous extracellular type II collagen. We evaluated the effects on four biomarkers associated with the creation of the denovo matrix using ELISA and immunohistochemistry: keratan sulfate epitope (5D4), 3B3(-) neoepitope of chondroitin-6- sulfate, 3B3(+) chondroitinase generated epitope of chondroitin-6-sulfate, and tenascin-C expression. TGF-beta1 stimulated the production of 3B3(+), 5D4, and tenascin-C in a dose-dependent manner and decreased 3B3(-) levels. Following the addition of exogenous type II collagen, 3B3(-) increased and tenascin-C decreased but did not change the direction of TGF-beta1 effects. In contrast, 5D4 expression decreased in the presence of collagen II as TGF-beta1 increased to 10 ng/ml. Interestingly, the amount of 3B3(+) epitope was not affected by the incorporation of type II collagen. Immunohistochemistry found there was no significant difference in distribution of these biomarkers in the presence and absence of extracellular type II collagen incorporation. These results elucidate the subtle biochemical differences in ECM synthesized by chondrocytes in the presence of type II collagen and further characterize the role played by ECM in the TGF-beta1 regulation of the articular cartilage physiology. PMID- 12568961 TI - Localization of insulin-like growth factor binding protein-2 in chondrocytes of bovine articular cartilage. AB - PURPOSE: Previous work indicated that transforming growth factor (TGF-beta) treatment of bovine articular cartilage resulted in an accumulation of insulin like growth factor binding protein-2 (IGF-BP-2). The purpose of the work presented in this paper was to define the localization of the IGF-BP-2 in freshly excised articular cartilage and in slices cultured in the presence and absence of TGF-beta. METHOD: Newborn calf articular cartilage was dissected and immediately fixed or maintained in organ culture for five days under basal conditions (media without added serum or growth factors) or with basal media containing 15 ng/ml of TGF-beta1. Frozen or paraffin embedded sections were prepared, and immunohistochemistry using anti-IGF-BP-2 performed. RESULTS: The paraffin sections provided the best preservation of morphology and consistency of immunohistochemical staining patterns. In fresh cartilage slices, IGF-BP-2 was associated with most of the chondrocytes. The basal cultured cartilage showed positive immunostaining in some areas, but not others: the most consistently stained area was the upper radial zone. In all cases where a positive reaction was observed, it was associated mostly with chondrocytes. On the other hand, all the TGF-beta treated samples that were examined in this study were evenly stained, and most chondrocytes were positive in all areas from superficial to deep zones, thus closely resembling the pattern of fresh tissue. CONCLUSIONS: It is concluded that IGF-BP-2 is closely cell associated in bovine articular cartilage. Following culture of cartilage slices, TGF-beta increases the number of cells with positive immunostaining. These data help to support the postulate that TGF-beta exerts at least some of its actions in articular cartilage via cross-talk mechanisms involving the IGF-BP-2 system. PMID- 12568962 TI - Induction of CD-RAP mRNA during periosteal chondrogenesis. AB - Induction of chondrogenesis and maintenance of the chondrocyte phenotype are critical events for autologous periosteal transplantation, which is a viable approach for cartilage repair. Cartilage-derived retinoic acid-sensitive protein (CD-RAP) is a recently discovered protein that is mainly produced in cartilage. During development, CD-RAP expression starts at the beginning of chondrogenesis and continues throughout cartilage maturation. In order to investigate the involvement of CD-RAP during periosteal chondrogenesis we have determined the nucleotide sequence of the rabbit CD-RAP mRNA and utilized this information to evaluate the temporal and spatial expression pattern of CD-RAP at the mRNA level during chondrogenesis. When the periosteal explants were cultured under chondrogenic conditions, the expression of CD-RAP was induced, as shown by a 40 fold increase in CD-RAP mRNA between days 7 and 10. The temporal expression pattern of CD-RAP closely mimicked that of collagen type IIB mRNA. Also, the CD RAP mRNA was localized to the matrix forming chondrocytes in the cambium layer of the periosteum by in situ hybridization as indicated by colocalization with collagen type II mRNA and positive safranin O staining. These data suggest a regulatory role of CD-RAP in periosteal chondrogenesis, which is potentially important for both cartilage repair and fracture healing via callus formation. PMID- 12568963 TI - The use of intra-articular Na-hyaluronate as a potential chondroprotective device in experimentally induced acute articular cartilage injury and repair in rabbits. AB - OBJECTIVE: This study examined if viscosupplementation from intra-articular administration of a commercially available form of hyaluronan (HA) could promote the restoration of proteoglycan (PG) depletion induced by chymopapain and then if the repair could be maintained once HA treatment was discontinued. METHODS: Animals received cartilage injury with intra-articular chymopapain (2.0 mg) followed by weekly treatment with intra-articular HA. HA treated animals were compared to injured animals with no treatment, contralateral untreated joints and joints from normal controls. The effect of intra-articular HA alone on articular cartilage was also examined. RESULTS: Serum keratan sulfate levels confirmed degradation of the cartilage PGs in the chymopapain-injected knees. Intra articular chymopapain resulted in marked loss of PGs. There were no significant differences among the control groups (untreated control, HA/800 treatment only). HA treatment did not affect the loss of PGs caused by chymopapain after 42 days. However, in animals receiving chymopapain injury followed by weekly HA treatment for 42 days and then 42 days of free cage activity without HA, cartilage PG contents were significantly increased. Intra-articular HA alone had no effect on the articular cartilage. CONCLUSION: The results in the present study suggest a potential protective effect of HA on chymopapain-induced acute articular cartilage injury in rabbits that, in time, permits damaged cartilage to resynthesize matrix PGs after the HA treatment is discontinued. PMID- 12568964 TI - Determination of cortical bone porosity and pore size distribution using a low field pulsed NMR approach. AB - The objective of this study was first to prove the concept of a low field pulsed nuclear magnetic resonance (NMR) process for assessing the cortical porosity and pore size distribution of human bone in vitro, and then to apply the technique to detect age-related changes of bone in these parameters. The Carr-Purcell-Meiboom Gill NMR spin echo train method is used to determine the porosity, and an inversion NMR spin-spin relaxation (T(2)) spectrum is used to assess the pore size distribution in cortical bone. Using these techniques, cortical porosity and pore size distribution of 19 specimens of human cadaveric bone, ranging from 16 to 89 years of age, were assessed. The NMR results were compared with the histomorphometric data of the same bone samples to verify the efficacy of the NMR approach. Moreover, a coefficient (surface relaxivity) relating the pore size to the T(2) relaxation time was determined empirically for the Haversian canals and the osteocytic lacunae. The results of this study demonstrate that the in vitro NMR approach using T(2) relaxation techniques can directly assess the porosity and pore size distribution (Haversian canals and osteocytic lacunae) in human cortical bone. In addition, this study indicates that the age-related changes in cortical porosity relate predominantly to Haversian canals, whereas the porosity of osteocytic lacunae appears to be independent of age. PMID- 12568965 TI - Mitogens are increased in the systemic circulation during bone callus healing. AB - The influence of mechanical tissue strain caused by flexible fracture fixation on the systemic occurrence of systemic mitogens during callus healing was investigated. For this purpose the mitogenic capacity and growth factor concentration of sera from patients undergoing fracture treatment were determined. Sera from 9 patients whose fractures had been stabilized by external fixation were collected before and during fracture treatment. The sera were added to cell culture media of the osteoblastic cell line SaOS-2. After 5-6 days cell proliferation was measured. Transforming growth factor-beta1 (TGF-beta1) and insulin-like growth factor-I (IGF-I) concentrations were analyzed in serum samples from different healing stages. STATISTICS: paired Wilcoxon-test. Sera from fracture patients decreased SaOS-2 proliferation in the first week after surgery (p<0.05) compared to sera obtained prior to surgery. In the fourth or fifth week proliferation increased significantly (p<0.03). The increased proliferation of the SaOS-2 cells was associated with elevated levels of TGF-beta and IGF-I (p<0.05). The higher mitogenic activity of sera suggests an increased level of circulating mitogens. In a previous study this increase had also been observed in patients during distraction osteogenesis treatment but not in patients with primary bone healing by a stable fixated plate. It is therefore assumed that their release from the fracture site is a consequence of mechanical stimulation by interfragmentary movement of fracture ends. PMID- 12568966 TI - Pulsed electromagnetic fields affect phenotype and connexin 43 protein expression in MLO-Y4 osteocyte-like cells and ROS 17/2.8 osteoblast-like cells. AB - Osteocytes, the predominant cells in bone, are postulated to be responsible for sensing mechanical and electrical stimuli, transducing signals via gap junctions. Osteocytes respond to induced shear by increasing connexin 43 (Cx43) levels, suggesting that they might be sensitive to physical stimuli like low-frequency electromagnetic fields (EMF). Immature osteoblasts exhibit decreased intercellular communication in response to EMF but no change in Cx43. Here, we examined long term effects of pulsed EMF (PEMF) on MLO-Y4 osteocyte-like cells and ROS 17/2.8 osteoblast-like cells. In MLO-Y4 cell cultures, PEMF for 8 h/day for one, two or four days increased alkaline phosphatase activity but had no effect on cell number or osteocalcin. Transforming growth factor beta-1 (TGF-beta 1) and prostaglandin E(2) were increased, and NO(2-) was altered. PEMFs effect on TGF-beta1 was via a prostaglandin-dependent mechanism involving Cox-1 but not Cox 2. In ROS 17/2.8 cells, PEMF for 24, 48 or 72 h did not affect cell number, osteocalcin mRNA or osteocalcin protein. PEMF reduced Cx43 protein in both cells. Longer exposures decreased Cx43 mRNA. This indicates that cells in the osteoblast lineage, including well-differentiated osteoblast-like ROS 17/2.8 cells and terminally differentiated osteocyte-like MLO-Y4 cells, respond to PEMF with changes in local factor production and reduced Cx43, suggesting decreased gap junctional signaling. PMID- 12568968 TI - Evaluation of femoral perfusion in a non-traumatic rabbit osteonecrosis model with T2*-weighted dynamic MRI. AB - We evaluated femoral perfusion in a non-traumatic rabbit serum sickness osteonecrosis (ON) model, using serial repetitive T2*-weighted (T2*W) dynamic magnetic resonance imaging (MRI) and investigated prediction of ON occurrence in early stages, comparing T2*W dynamic MRI with non-enhanced (T2-, T1- and fat suppression T1-weighted) and contrast-enhanced MRI. Early microcirculatory injury or necrotic lesion was detected in 0% of femora (extravasation, 0/6) at 72 h, 33% (necrotic, 4/12) at 1 week and 100% (necrotic, 14/14) at 3 weeks using non enhanced MRI, and in 67% of femora (extravasation, 4/6) at 72 h, 58% (necrotic, 7/12) at 1 week and 100% (necrotic, 14/14) at 3 weeks using contrast-enhanced MRI. In contrast, microcirculatory injury or necrotic lesion was detected in 83% of femora (extravasation, 5/6) at 72 h, 92% (necrotic, 11/12) at 1 week and 100% (necrotic, 14/14) at 3 weeks using T2*W dynamic MRI as no transient decrease or less marked transient decrease in signal intensity of regions of interest (ROIs), compared to normal femora, which showed a clear transient decrease in signal intensity of ROIs. These results indicate that T2*W dynamic MRI with optimal imaging parameters and a dose of contrast agent is the most sensitive of these three MRI methods and may be clinically useful for evaluating femoral perfusion in artery phase and predicting ON occurrence. PMID- 12568967 TI - Effects of locally applied vascular endothelial growth factor (VEGF) and VEGF inhibitor to the rabbit tibia during distraction osteogenesis. AB - INTRODUCTION: Therapeutic angiogenesis, a novel concept in tissue engineering, is neo-formation of blood vessels in a tissue upon delivery of an angiogenic growth factor to the tissue. We hypothesised that therapeutic angiogenesis could enhance bone formation and challenged the hypothesis in an experimental model of distraction osteogenesis. METHODS: Rabbits, divided into three equal groups of 12, had their right tibia lengthened by distraction osteogenesis. A mini-osmotic pump delivered to the osteotomy gap either recombinant human vascular endothelial growth factor (VEGF), VEGF-inhibitor, or vehicle alone during the latency and distraction phase. After consolidation, we assessed bone blood flow by radioactive microsphere entrapment, measured torsional stiffness and bone mineral content, and did histomorphometry. RESULTS: VEGF and VEGF-inhibitor treatment failed to influence bone blood flow, torsional stiffness, bone mineral content and histomorphometric indices of the bone regenerate. However, VEGF treatment increased the blood flow in bone of the distracted limb and VEGF-inhibitor treatment decreased bone blood flow. CONCLUSION: The regenerate was unresponsive to VEGF and VEGF-inhibitor treatment in contrast to the neighbouring bone, which implies different biological properties of the vasculature in native and regenerating bone. VEGF is not recommended for enhancement of bone formation in this setting. PMID- 12568969 TI - A dissection and computer tomograph study of tarsal coalitions in 100 cadaver feet. AB - Most of the clinical studies report the incidence of tarsal coalitions (TC) as less than 1% but they disregard the asymptomatic coalitions. TC have been associated with degenerative arthritic changes. After X-rays, computer tomography (CT) is the most commonly used diagnostic test in the detection of TC. The aims of our study were to establish the incidence of TC; the association between TC and accessory tarsal bones and between TC and tarsal arthritis; and to assess the sensitivity of CT as a diagnostic tool in TC. We performed spiral CT scans of 100 cadaver feet (mean age at death 77.7+/-10.4), which were subsequently dissected. The dissections identified nine non-osseous TC: two talocalcaneal and seven calcaneonavicular. There was no osseous coalition. Tarsal arthritis was identified in 31 cases. Both talocalcaneal coalitions were associated with arthritis while none of the calcaneonavicular coalitions were associated with tarsal arthritis. The CT diagnosed an osseous talocalcaneal coalition and was suspicious of fibrocartilaginous coalitions in eight cases. There was correlation between dissection and CT in two talocalcaneal coalitions and three calcaneonavicular coalitions thus CT identifying 55.5% of the coalitions. CT did not diagnose four non-osseous coalitions and diagnosed errouresly four possible coalitions. In conclusion our study demonstrated that the incidence of non osseous TC is higher than previously thought (12.72%). The calcaneonavicular coalitions are the most common single type (9.09%) and they do not seem to be associated with arthritic changes in the tarsal bones. Our CT results suggest that spiral CT has a low sensitivity in the detection of non-osseous coalitions and questions if multislice CT should be used routinely when TC are suspected. PMID- 12568970 TI - Acute changes in the axonal cytoskeleton after mild stretching of the rat brachial plexus. AB - We have developed an animal model to investigate acute changes in the axonal cytoskeleton caused by a mild stretching of the peripheral nerve in the upper limbs of rats. Rat forelimbs were continuously stretched at 2 N for 1 h. Thereafter, a part of the brachial plexus and median nerve were harvested and processed for electron microscopic analysis. The total number of microtubules in the brachial plexus decreased to 55% of that of the control animals (p<0.05) without change in the number of neurofilaments. No significant changes in microtubules or neurofilaments were observed in the median nerve. By Western blotting analysis, the amount of tau protein in the stretch group significantly decreased in the brachial plexus but not in the median nerve. However, no significant changes in the amount of tubulin protein were observed in either the brachial plexus or median nerve. These results suggest that the microtubules were depolymerized by stretching of the brachial plexus and that the depolymerization may have been mediated by the decrease in the tau protein. PMID- 12568971 TI - Spinal somatosensory evoked potential evaluation of acute nerve-root injury associated with pedicle-screw placement procedures: an experimental study. AB - Pedicle screws for spinal fixation risk neural damage because of the proximity between screw and nerve root. We assessed whether spinal somatosensory evoked potential (SSEP) could selectively detect pedicle-screw-related acute isolated nerve injury. Because pedicle screws are too large for a rat's spine, we inserted a K-wire close to the pedicle in 32 rats, intending not to injure the nerve root in eight (controls), and to injure the L4 or L5 root in 24. We used sciatic-nerve elicited SSEP pre- and postinsertion. Radiologic, histologic, and postmortem observations confirmed the level and degree of root injury. Sciatic (SFI), tibial (TFI), and peroneal function indices (PFI) were calculated and correlated with changes in potential. Although not specific for injuries to different roots, amplitude reduction immediately postinsertion was significant in the experimental groups. Animals with the offending wire left in place for one hour showed a further non-significant deterioration of amplitude. Electrophysiologic changes correlated with SFI and histologic findings in all groups. SSEP monitoring provided reliable, useful diagnostic and intraoperative information about the functional integrity of single nerve-root injury. These findings are clinically relevant to acute nerve-root injury and pedicle-screw insertion. If a nerve-root irritant remains in place, a considerable neurologic deficit will occur. PMID- 12568972 TI - Influence of a standardized closed soft tissue trauma on resistance to local infection. An experimental study in rats. AB - PURPOSE: The etiology of local posttraumatic infection in the locomotor system depends on the amount, virulence and pathogenicity of the inoculated microorganisms and the local/systemic host damage due to the type and extent of the accident or iatrogenic trauma. The relative effect of these factors remains unclear. In particular, it is still unclear today whether--in presence of microorganisms--soft tissue damage and its pathophysiological consequences lead to infection after soft tissue trauma, or whether the bacterial contamination is the primarily cause for posttraumatic infection. The aim of the project was to gain information on the consequences of a soft tissue injury in terms of resistance to local infection. Since clinical populations are too heterogeneous, the problem was investigated in a standardized, reduced (no surgery or implants) experimental in vivo model. METHOD: In female Sprague-Dawley-rats with a standardized closed soft tissue trauma to the tibialis anterior muscle (group I: n=13) or without (group II: n=13), we compared the incidence of local infection after a pairwise local, percutaneously injected bacterial challenge with various concentrations of Staphylococcus aureus (2 x 10(4)-2 x 10(6) colony forming units, CFU). The standardized closed soft tissue trauma was created by application of a specially designed, computer controlled impact device. The contaminated soft tissue and the underlying bone were removed under sterile conditions after five days and quantitatively evaluated for bacterial growths. Infection was defined as positive bacterial growth at the soft tissue and/or bone. A stepwise experimental design with an "up-and-down" dosage technique was used to adjust the bacterial challenge in the area of the ID50 (50% infection dose). Statistical evaluation of the difference between the infection rates of both groups was performed by two-sided fisher exact test (p<0.05). RESULTS: The overall infection rate was 46%. For the group with soft tissue trauma the ID50 was 1.32 x 10(5) CFU and 1.05 x 10(6) CFU for the group without soft tissue trauma. The infection rate was 69% (9 of 13 animals) for the group with soft tissue trauma and 23% (3 of 13 animals) for the group without soft tissue trauma. This difference is statistically significant (p=0.047). CONCLUSIONS: The infection rate after a standardized closed soft tissue injury was significantly higher and the ID50 lower than without soft tissue trauma. Our results demonstrate that in presence of microorganisms it is not primarily the bacterial contamination but rather the soft tissue damage and its pathophysiological consequences resulting in decreased infection resistance that secondarily lead to infection. PMID- 12568973 TI - Direct costs for the treatment of HIV-infection in a German cohort after the introduction of HAART. AB - BACKGROUND: Highly active antiretroviral combination therapy (HAART) has become the standard of care for HIV infection. The reduction of morbidity by HAART has been proven to be cost-effective despite high expenditures for regular use of antiretrovirals. We examined direct costs in a German monocentric cohort of HIV infected patients after introduction of HAART. SUBJECTS/METHODS: In 1997 recruitment started and 201 patients gave informed consent. They underwent structurized interviews. Additional data were taken from the patients records. Later on follow-ups were performed for the same cohort in the years 2000 and 2001 respectively. Direct costs have been calculated per patient and year for each period. RESULTS: The proportion of HAART treated patients rised in the cohort from 86% to 93%. The mean of antiretrovirals used per case increased from 2.4 to 3.4. Nevertheless mean direct costs for HAART decreased significantly from Euro 17,746 to Euro 16,007. Reduction of expenditures for additional drugs, hospitalisation and diagnostics led to about one third decrease of mean total direct cost from Euro 35,865 in 1997 to Euro 24,482 in 2001. For surviving patients expenditures remained higher in advanced stage of disease for HAART, hospitalisation, diagnostics and total costs. CONCLUSION: Expenditures for HAART remained on a high level. Despite rising drug prices and increased use of antiretrovirals a decrease of mean costs of HAART by about 10% resulted from more frequent use of less expensive drug combinations. The continuous decrease of expenditures for non-HAART drugs, diagnostics and hospitalisation predominated and therefore HAART caused about a half of total direct costs in 1997 and two third in 2001 respectively. Higher expenditures in advanced stages of disease continued over the follow up period and raise the question of an economic impact of earlier initiation of treatment. More extensive use of standardized evaluation of direct costs could be an important tool towards a more rational allocation of resources in health care. PMID- 12568974 TI - Immunoglobulin deposition in sural nerves of AIDS patients with distal-symmetric HIV-associated polyneuropathy. AB - The most frequent neurological diagnosis in peripheral nerve function of HIV positive individuals is distal-symmetric polyneuropathy (DSPN). In this study we investigated the histopathology as well as the immunohistochemical expression of immunoglobulins IgA, IgG and IgM in post-mortem sural nerve tissue gained from 11 patients who had suffered from DSPN in the clinical course of AIDS (CDC 3C). We found that all 11 sural nerves showed signs of demyelination while in 6 out of 11 cases axonal degeneration could also be detected. Immunohistochemical expression of at least one immunoglobulin was found in all but two cases with deposits uniformly being located immediately beneath the basement membrane of capillary blood vessels and within the perineurium while endoneurial staining was discernable in three cases. The most commonly expressed immunoglobulin was IgA which was identified in 7 cases, followed by IgG and IgM which were positive in 6 and 5 cases, respectively. All three immunoglobulins were found to be expressed simultaneously in only two cases. Thus, our study shows that immunoglobulin deposits among other factors may be implicated in altering the function of sural nerves or enhance their vulnerability. In peripheral nerves they may be responsible for some of the common alterations in the development of AIDS associated distal symmetric polyneuropathy. PMID- 12568975 TI - Remission of a cutaneous Mycosis fungoides after topical 5-ALA sensitisation and photodynamic therapy in a patient with advanced HIV-infection. AB - BACKGROUND: Treatment of Mycosis fungoides (MF) in HIV-infected patients is controversially discoursed. Photodynamic therapy (PDT) after topical sensitization with 5-aminolevulinic acid (5-ALA) is a new and effective modality for treatment of skin malignancies. OBJECTIVE: In this report we describe, what is, to our knowledge, the first case of a patient with MF through advanced HIV infection, successfully experiencing topical 5-ALA sensitization and PDT. METHODS: 5-ALA ointment was applied to plaques and held in occlusion for 4 hours. PDT was applied using the PDT 1200 irradiation source (Waldmann Medizintechnik System) with 180 J/cm superset 2. RESULTS: Complete remission of MF was achieved, after two completed cycles of photodynamic therapy. CONCLUSION: MF lesions in the presended case showed a high response to 5-ALA sensitization and PDT. This modality appeared to be very effective in treatment of MF in a HIV-infected patient and could be a valuable treatment option for cutaneous T-cell lymphoma in HIV-infected patients. PMID- 12568976 TI - Efficacy and tolerability of valerian extract LI 156 compared with oxazepam in the treatment of non-organic insomnia--a randomized, double-blind, comparative clinical study. AB - Patients aged 18 to 73 years and diagnosed with non-organic insomnia according to ICD-10 (F 51.0) were treated in a multicentre, double-blind, randomised parallel group comparison with either 600 mg/die valerian extract LI 156 (Sedonium) or 10 mg/die oxazepam taken for 6 weeks. A total of 202 outpatients with a mean duration of insomnia of 3.5 months at baseline were included at 24 study centres (general practices) in Germany. - Sleep quality (SQ) after 6 weeks measured by the Sleep Questionnaire B (SF-B; CIPS 1996) showed that 600 mg/die valerian extract LI 156 was at least as efficacious as a treatment with 10 mg/die oxazepam. Both treatments markedly increased sleep quality compared with baseline (p <0.01). The other SF-B subscales, i.e. feeling of refreshment after sleep (GES), psychic stability in the evening (PSYA), psychic exhaustion in the evening (PSYE), psychosomatic symptoms in the sleep phase (PSS), dream recall (TRME), and duration of sleep confirmed similar effects of both treatments. Clinical Global Impressions scale (CGI) and Global Assessment of Efficacy by investigator and patient, again, showed similar effects of both treatments. Adverse events occurred in 29 patients (28.4%) receiving valerian extract LI 156 and 36 patients (36.0%) under oxazepam, and were all rated mild to moderate. No serious adverse drug reactions were reported in either group. Most patients assessed their respective treatment as very good (82.8% in the valerian group, 73.4% in the oxazepam group). During the 6 week treatment phase Valerian extract LI 156 (Sedonium) 600 mg/die showed a comparable efficacy to 10 mg/die oxazepam in the therapy of non-organic insomnia. PMID- 12568977 TI - Elevation of soluble tumor necrosis factor receptor II in non-febrile patients with acute myeloid leukemia. AB - BACKGROUND: Tumor necrosis factor alpha (TNF) and its cellular and soluble (s) receptors (TNF-R) are important mediators in acute myeloid leukemia (AML) and infectious complications during cytoreductive therapy. We investigated the serum concentrations of sTNF-RII in previously untreated patients with AML at the onset of cytoreductive therapy and in non-febrile chemotherapy-associated neutropenia. PATIENTS AND METHODS: Of 54 eligible patients with AML, serum concentrations of sTNF-RII could be evaluated in 25 non-neutropenic, non-febrile and in 11 neutropenic, non-febrile patients. RESULTS: At baseline, non-neutropenic, non febrile AML patients showed high median serum sTNF-RII concentrations of 3,804 pg/mL. In neutropenia, there was a non-significant trend (p = 0.18) to lower median sTNF-RII levels of 3,246 pg/mL. CONCLUSIONS: Serum sTNF-RII concentrations in non-febrile AML patients before chemotherapy are in the range of levels reached in uncomplicated febrile episodes in otherwise healthy individuals. This must be taken into account when evaluating the cytokine profile for sepsis in patients with therapy-associated neutropenia. Concentrations are still elevated in neutropenia, suggesting that a normal number of leukocytes is not necessarily required for the activation of the TNF ligand/TNF receptor system in AML. PMID- 12568978 TI - Evaluation of sulcular sulphide level monitoring using a portable sensor system. AB - PURPOSE: Volatile sulphur compounds (VSC) such as hydrogen sulphide and methyl mercaptan are toxic bacterial metabolites considered to contribute to the progression of periodontitis. However, the quantitative determination of sulcular VSC concentration requires costly time-consuming laboratory methods limiting its evaluation in large clinical trials. Recently, a portable monitor with a sulphide microsensor has been developed to measure sulcular VSC levels (SU). This cross sectional study evaluated 1) the clinical usage of the sulphide sensor system, 2) the reproducibility of SU-measurements, 3) the distribution of SU-values in periodontal health and disease and 4) the relationship between SU and clinical parameters documenting periodontal disease severity. METHODS: The specificity, the measuring range and the reproducibility of the sensor system were tested with reference solutions in vitro. 10 probands with healthy gingiva, 23 patients with gingivitis and 30 patients with chronic periodontitis were examined recording periodontal parameters (clinical attachment loss, CAL, probing depth, PD, bleeding on probing intensity BI, plaque and gingival index PI, GI) and sulcular sulphide level (SU) measured by the portable monitor as a digital score ranging from 0.0 (< 10 superset -6 M of S2-) to 5.0 (10 superset -2 M of S superset 2-) in increments of 0.5. RESULTS: The in vitro testing confirmed the sensor s specificity for sulphide ions. The intraexaminer agreement (Pearson correlation coefficient) was 0.91 between two measurements (p < 0.05). However, positive SU values decreased with measurement repetition due to the volatility of the sulphur compounds. SU and the percentage of positive SU-values per patient were correlated with disease severity. There were no positive SU-values in healthy probands, a mean SU of 0.11 +/- 0.08 in gingivitis and of 0.22 +/- 0.15 in periodontitis (p <0.001). SU was significantly correlated with PD in gingivitis, PD and CAL in periodontitis and the bleeding index in the analysis of all data. CONCLUSION: The portable sulphide monitor is a reliable method for the chairside determination of sulcular sulphide level in periodontal disease and in spite of its limitations a valuable supplementation of traditional clinical examination methods. It offers the possibility of relating in vitro results on the toxicity of VSC to clinical findings and might provide new insight into the impact of sulcular sulphide on the pathophysiology of periodontitis. PMID- 12568979 TI - Effect of C-peptide on wound healing and microcirculation in diabetic mice. AB - AIM: Recent studies have demonstrated that C-peptide is biologically active and might have a beneficial effect on late complications in diabetes mellitus. The aim of this study was to investigate the effects of systemically given C-peptide on dermal wound healing in diabetic mice. METHODS: Experiments were carried out in male SKH-1 hr hairless mice. Dermal wounds were created (diameter 2.5mm) in streptozotozin-diabetic and normal control mice. Mice were randomized into three treatment groups (n = 10 each): Normal controls, diabetic mice with PBS or C peptide injection twice daily. At various time points (prior wounding as well as days 4, 7, 10 and 15) microcirculation was quantitatively analyzed by intravital fluorescent microscopy to determine wound surface area, vessel diameter, red blood cell velocity, plasma leakage, functional capillary density. In addition, leukocyte/endothelium interaction was quantified by in vivo visualization of leukocytes. RESULTS: Systemic administration of C-peptide showed no influence on wound healing or standard microcirculatory parameters. The leukocyte/ endothelium interaction revealed a significant (p<0.05) increase in the number of adherent leukocytes 15 days after wound creation in C-peptide treated diabetic mice. CONCLUSION: Except for the significantly increased number of leukocytes adherent to venular endothelium in the C-peptide group no alteration was observed in wound healing and microcirculation. Neutrophil recruitment after C-peptide injection is of interest because it may reduce the risk of infection in diabetes mellitus. PMID- 12568982 TI - BM-derived stem cells for the treatment of nonhematopoietic diseases. PMID- 12568983 TI - Multipotential stem cells and 'side population' cells. PMID- 12568984 TI - Adipose tissue: challenging the marrow monopoly. PMID- 12568985 TI - On the development of cell therapy for genetic disorders. PMID- 12568986 TI - Muscular dystrophies and stem cells: a therapeutic challenge. PMID- 12568987 TI - Adult BM stem cells regenerate mouse myocardium. PMID- 12568988 TI - Mesenchymal 'stem cell rescue' for myocardial disease. PMID- 12568989 TI - GM-CSF versus G-CSF: engraftment characteristics, resource utilization, and cost following autologous PBSC transplantation. AB - BACKGROUND: G-CSF and GM-CSF have both been shown to decrease the time to hematopoietic recovery when administered after autologous BM or peripheral stem cell re-infusion. However, few studies have compared G-CSF and GM-CSF to determine which is the preferred myeloid growth factor. METHODS: This study compares a prospectively accrued cohort of 22 patients receiving GM-CSF with a historical cohort of patients who received G-CSF commencing Day + 6 after autologous PBSC transplantation. Patients were matched based on disease type and stage, CD34(+) cell dose/kg, conditioning regimen, and prior treatment. Time to myeloid engraftment, growth factor utilization, antibiotic utilization, fever incidence, and cost were compared. RESULTS: The median time to neutrophil and platelet engraftment was similar in the two groups (ANC > 500 /mm(3), GM-CSF 12 versus G-CSF 11, P = 0.69). There was a trend towards more days of temperature > 38.0 masculine C (six versus three, P = 0.05) and febrile neutropenia (three versus two, P = 0.06) in the GM-CSF arm. There was a trend towards increased use of i.v. antibiotics in the GM-CSF cohort (7.6 days versus 5.5 days, P = 0.06). More chest X-rays (1.5 versus 1.0, P = 0.03) were ordered, and more blood cultures drawn (4.2 versus 2.7, P = 0.05) as part of fever evaluation in the group treated with GM-CSF. Resource utilization based on actual wholesale pricing (AWP) favored the G-CSF cohort. Applying a sensitivity analysis, GM-CSF became cost-effective when priced below $94 per 250 micro g, despite greater resource utilization. DISCUSSION: This study suggests that engraftment characteristics are similar with GM-CSF and G-CSF following PBSC transplantation. Resource utilization for fever treatment and evaluation may be greater with GM-CSF. Determination of which agent is more cost-effective depends on institutional acquisition costs. PMID- 12568990 TI - Catastrophic failures of freezing bags for cellular therapy products: description, cause, and consequences. AB - BACKGROUND: Container integrity is critical for maintaining sterility of cryopreserved cellular therapy products. We investigated a series of catastrophic bag failures, first noticed in early 2001. METHODS: Process records were reviewed for all PBPC and lymphocyte products cryopreserved in bags from January 2000 through April 2002. Patient charts were also reviewed. RESULTS: One thousand two hundred and four bags were removed from storage for infusion to 261 patients. All products had been cryopreserved in Cryocyte poly(ethylene co-vinyl acetate) (EVA) bags in either 10% DMSO or 5% DMSO and 6% pentastarch. Product volumes were 25-75 mL, and bags were stored with overwrap bags in a liquid nitrogen tank. From January 2000 to April 2001, failure occurred in 10 of 599 (1.7%) bags. From May 2001 to April 2002, 58 of 605 (9.6%) bags failed, typically with extensive fractures that were visible before thaw. Of the 58 that failed, 24 were salvaged by aseptic methods and infused to patients under antibiotic coverage; 10 of those 24 (42%) had positive bacterial cultures. Bag failures were not related to product type, cryoprotectant solution, liquid versus vapor storage, or freezer location. Failures were linked to use of four Cryocyte bag lots manufactured in 2000 and 2001. After replacing these lots with a 1999 Cryocyte lot and with KryoSafe polyfluoroethylene polyfluoropropylene (FEP) bags, no more failures occurred in 75 and 102 bags, respectively, thawed through April 2002. DISCUSSION: High rates of bag failure were associated with four Cryocyte bag lots. No serious adverse patient effects occurred, but bag failures led to microbial contamination, increased product preparation time, increased antibiotic use, and increased resource expenditure to replace products. PMID- 12568991 TI - Thawing of cryopreserved mobilized peripheral blood--comparison between waterbath and dry warming device. AB - BACKGROUND: Thawing of cryopreserved mobilized peripheral blood (MPB) is routinely performed for autologous and allogeneic MPB transplantation. Usually thawing is achieved by submerging the cell bag in a waterbath (37 degrees C temperature). We compared the effectiveness of thawing cryopreserved MPB in a waterbath with an electric dry-warming device containing warmed gel pads (Sahara, Transmed). METHODS: Two cryopreserved bags from each of 31 apheresis procedures were thawed in a waterbath and under dry conditions in parallel. Viability (dye exclusion), apoptosis/necrosis (annexin/propidiumiodide staining) and clonogenic potential (CFU-E plus BFU-E, CFU-GM) of the cells were tested after thawing. RESULTS: Statistical analysis by Wilcoxon matched-pair test showed no significant difference between the thawing procedures in terms of the in vitro parameters tested. DISCUSSION: Our results indicate that thawing of cryopreserved MPB using dry warming and water bath give similar viability, apoptosis/necrosis rate and clonogenic potential. Both procedures take about the same amount of time and are easy to perform. Nevertheless, the potentially decreased risk of bacterial contamination of either the cell product or the patient room, and guidelines of good clinical practice (GCP), favor the use of the dry warming procedure. PMID- 12568992 TI - False positive endotoxin results in a DC product caused by (1-->3)-beta-D-glucans acquired from a sterilizing cellulose filter. PMID- 12569011 TI - Single-hit mechanism of tumour cell killing by radiation. AB - PURPOSE: To review the relative importance of the single-hit mechanism of radiation killing for tumour response to 1.8-2.0 Gy day(-1) fractions and to low dose-rate brachytherapy. MATERIALS AND METHODS: Tumour cell killing by ionizing radiation is well described by the linear-quadratic equation that contains two independent components distinguished by dose kinetics. Analyses of tumour cell survival curves that contain six or more dose points usually provide good estimates of the alpha- and beta-inactivation coefficients. Superior estimates of tumour cell intrinsic radiosensitivity are obtained when synchronized populations are employed. The characteristics of single-hit inactivation of tumour cells are reviewed and compared with the characteristics of beta-inactivation. Potential molecular targets associated with single-hit inactivation are discussed along with strategies for potentiating cell killing by this mechanism. RESULTS: The single-hit mechanism of tumour cell killing shows no dependence on dose-rate and, consequently, no evidence of sublethal damage repair. It is uniquely potentiated by high linear-energy-transfer radiation, exhibits a smaller oxygen enhancement ratio and exhibits a larger indirect effect by hydroxyl radicals than the beta mechanism. alpha-inactivation coefficients vary slightly throughout interphase but mitotic cells exhibit extremely high alpha-coefficients in the range of those observed for lymphocytes and some repair-deficient cells. Evidence is accumulating to suggest that chromatin in compacted form could be a radiation hypersensitive target associated with single-hit radiation killing. CONCLUSIONS: Analyses of tumour cell survival curves demonstrate that it is the single-hit mechanism (alpha) that determines the majority of cell killing after doses of 2Gy and that this mechanism is highly variable between tumour cell lines. The characteristics of single-hit inactivation are qualitatively and quantitatively distinct from those of beta-inactivation. Compacted chromatin in tumour cells should be further investigated as a radiation-hypersensitive target that could be modulated for therapeutic advantage. PMID- 12569012 TI - Chromosomal biodosimetry by unfolding a mixed Poisson distribution: a generalized model. AB - PURPOSE: The current chromosomal biodosimetry system is practically limited to the estimation of equivalent whole-body dose in the low or moderate dose range. To circumvent this problem, a new dosimetry system was developed which was effective over a wide dose range and dose gradient. MATERIALS AND METHODS: In order to establish a quantitative formalism of a comprehensive dose-response kinetics, chromosome aberration frequencies were studied in human peripheral blood lymphocytes irradiated in vitro with gamma-rays in a dose range of 0.01 50Gy. Using the dose-yield relationship thus established, a new model of biodosimetry was developed which involved unfolding the chromosome aberration distribution into a mixed Poisson distribution and thence into a dose distribution profile. The model was then tested with chromosome aberration data in lymphocytes irradiated in vitro, simulated partial body irradiation, accidental radiation exposure, therapeutic local irradiation and protracted exposure by internal deposit of Thorotrast. RESULTS: The dose-yield relations over a wide dose range fit satisfactorily a multiparametric dose-response curve, which included the first approximation kinetics of linear-quadratic dose-response and its distortion by mutual interference among multiple breaks on a chromosome arm. The unfolding strategy using these dose-response kinetics was successfully applied to the various types of radiation exposures; the reconstructed dose profile showing a sharp unimodal peak for moderate or high doses of gamma-rays in vitro irradiation, broadening for high-linear energy transfer radiation, discrimination of doses in the mixed culture of irradiated and unirradiated blood, and reasonable dose distribution for the in vivo exposures. CONCLUSION: A novel biodosimetry system was developed. The system is innovative in that it provides information not only on the macroscopic dose inhomogeneity, but also on the microscopic spectrum of doses stemming from the variability of energy transfer by charged particles as well as the multiple ionization events to which the target cell nucleus is exposed. PMID- 12569013 TI - Effects of cell cycle phase on low-dose hyper-radiosensitivity. AB - PURPOSE: To examine the low-dose radiation response of human glioma cell lines separated into different cell-cycle phases and to determine if low-dose hyper radiosensitivity (HRS) differs in populations defined by cell-cycle position. To assess whether predictions of the outcome of multiple low-dose regimens should take account of cell-cycle effects. MATERIALS AND METHODS: The clonogenic survival of G1, G2 and S phase cells was measured after exposure to single doses of X-rays in two human glioma cell lines. One cell line (T98G) showed marked HRS when asynchronous cells were irradiated, while the other (U373) did not. Separation of populations and high-resolution cell counting was achieved using a fluorescence activated cell sorter. Sorted cell populations were irradiated with 240 kVp X-rays to doses between 0.05 and 5Gy. The resulting cell-survival versus dose data were comparatively fitted using the linear-quadratic and induced-repair models in order to assess the degree of HRS. RESULTS: In both cell lines the low dose response was altered when different populations were irradiated. In T98G cells, all populations showed HRS, but this was most marked in G2 phase cells. In U373 cells, no HRS was found in G1 or S phase cells, but HRS was demonstrable in G2 phase cells. CONCLUSIONS: HRS was expressed by the whole cell population of T98G cells but the size of the effect varied with cell-cycle phase and was most marked in the G2 population. In U373 cells, the effect could only be demonstrated in G2 cells. This implies that HRS is primarily a response of G2 phase cells and that this response dominates that seen in asynchronous populations. Actively proliferating cell populations may therefore demonstrate a greater increase in radiosensitivity to very low radiation doses compared with quiescent populations. PMID- 12569014 TI - Relative biological effectiveness (RBE) of 210Po alpha-particles versus X-rays on lethality in bovine endothelial cells. AB - PURPOSE: Alpha-radiation from polonium-210 ((210)Po) can elevate background radiation dose by an order of magnitude in people consuming large quantities of meat and seafood, particularly caribou and reindeer. Because up to 50% of the ingested (210)Po body burden is initially found in the blood, a primary target for the short range alpha-particles is the endothelial cells lining the blood vessels. This study examined the relative biological effectiveness (RBE) of (210)Po alpha-particles versus 250 kVp X-rays in producing injury to cultured bovine aortic endothelial cells. MATERIALS AND METHODS: Radiation effects on cells were measured in four different ways: the percentage viable cells by trypan blue dye exclusion, the number of live cells, the lactate dehydrogenase (LDH) release to medium and the ability to form colonies (clonogenic survival). RESULTS: Comparison of dose-response curves yielded RBE values of 13.1+/-2.5 (SEM) for cell viability, 10.3+/-1.0 for live cell number and 11.1+/-3.0 for LDH activity. The RBE values for clonogenic survival were 14.0+/-1.0 based on the ratio of the initial slopes of the dose-response curves and 13.1, 9.9 and 7.7 for 50, 10 and 1% survival rate, respectively. At X-ray doses <0.25 Gy, a pronounced stimulatory effect on proliferation was noted. CONCLUSIONS: Exposure to (210)Po alpha-particles was seven to 14 times more effective than X-ray exposure in causing endothelial cell damage. PMID- 12569015 TI - Effect of keratinocyte growth factor on the proliferation, clonogenic capacity and colony size of human epithelial tumour cells in vitro. AB - PURPOSE: The effect of recombinant human keratinocyte growth factor (rHuKGF) on the proliferation, clonogenic capacity and colony size of low-passage human epithelial tumour cells was tested in vitro. MATERIALS AND METHODS: Five tumour cell cultures derived from head and neck squamous cell carcinomas, three cultures derived from pleural effusions of carcinomas of different origin and normal human nasal epithelial cells were analysed in passages 2-4. Expression of FGF7 and its receptor (FGFR2) were determined by the RNase protection assay. Cells were incubated with rHuKGF (10-200 ng ml(-1)) 3 days before or immediately after plating for clonal growth in serum-depleted media. To determine cellular radiosensitivity, single doses of 1-8 Gy X-rays were applied. Colony formation as well as colony size, reflecting the number of cell divisions, was determined after 10-15 days of growth in rHuKGF-treated and control cells. RESULTS: Normal nasal epithelial cells showed a two- to threefold increase in the number of cell divisions due to rHuKGF-treatment. In tumour cell cultures, significant stimulation of proliferation occurred in only one of eight samples. Tumour cells expressed FGF7 mRNA and protein, and low levels of FGFR2 mRNA. The addition of rHuKGF to the medium of the tumour cell cultures influenced neither radiation induced impairment of proliferation nor clonogenic cell survival. CONCLUSION: rHuKGF has been shown to ameliorate the radiation tolerance of normal epithelia. The minimum in vitro tumour cell response to rHuKGF compared with normal epithelial cells suggests a potential for selective protection of normal epithelia during radiotherapy. The low FGFR2 expression as well as the FGF7 expression in the tumour cells may contribute to their resistance to rHuKGF treatment. PMID- 12569016 TI - Radiation dose-dependent increases in inflammatory response markers in A-bomb survivors. AB - PURPOSE: The well-documented increases in malignant tumours in the A-bomb survivors have recently been supplemented by reports that non-cancer diseases, including cardiovascular disease, may also have increased in incidence with increasing radiation dose. Given that low-level inflammatory responses are widely accepted as a significant risk factor for such diseases, we undertook a detailed investigation of the long-term effects of ionizing radiation on the levels of the inflammatory markers C-reactive protein (CRP) and interleukin 6 (IL-6) in A-bomb survivors. MATERIALS AND METHODS: Blood samples were taken from 453 participants in a long-term epidemiological cohort of A-bomb survivors. Plasma levels of CRP and IL-6 were measured using standard antibody-mediated procedures. Relationships between CRP or IL-6 levels and radiation dose were then investigated by multivariate regression analysis. Blood lymphocytes from each individual were used for immunophenotyping by flow cytometry with murine monoclonal antibodies to CD3, CD4 and CD8. RESULTS: CRP levels were significantly increased by about 31% Gy(-1) of estimated A-bomb radiation (p=0.0001). Higher CRP levels also correlated with age, male gender, body mass index and a history of myocardial infarction. After adjustments for these factors, CRP levels still appeared to have increased significantly with increasing radiation dose (about 28% increase at 1Gy, p=0.0002). IL-6 levels also appeared to have increased with radiation dose by 9.3% at 1Gy (p=0.0003) and after multiple adjustments by 9.8% at 1Gy (p=0.0007). The elevated CRP and IL-6 levels were associated with decreases in the percentages of CD4(+) helper T-cells in peripheral blood lymphocyte populations. CONCLUSIONS: Our results appear to indicate that exposure to A-bomb radiation has caused significant increases in inflammatory activity that are still demonstrable in the blood of A-bomb survivors and which may lead to increased risks of cardiovascular disease and other non-cancer diseases. PMID- 12569017 TI - Association of transforming growth factor beta-1 single nucleotide polymorphisms with radiation-induced damage to normal tissues in breast cancer patients. AB - PURPOSE: To investigate whether transforming growth factor beta-1 (TGFbeta1) single nucleotide polymorphisms were associated with the susceptibility of breast cancer patients to severe radiation-induced normal tissue damage. MATERIALS AND METHODS: PCR-RFLP assays were performed for TGFbeta1 gene polymorphisms on DNA obtained from 103 breast cancer patients who received radiotherapy. The G-800A, C 509T, T+869C and G+915C polymorphic sites were examined, and genotype and allele frequencies of two subgroups of patients were calculated and compared. RESULTS: The less prevalent -509T and +869C alleles were significantly associated with a subgroup of patients who developed severe radiation-induced normal tissue fibrosis (n=15) when compared with those who did not (n=88) (odds ratio=3.4, p=0.0036, and 2.37, p=0.035, respectively). Furthermore, patients with the -509TT or +869CC genotypes were between seven and 15 times more likely to develop severe fibrosis. CONCLUSIONS: These findings imply a role for the -509T and +869C alleles in the pathobiological mechanisms underlying susceptibility to radiation induced fibrosis. Their predictive value would be limited to patients who are 509TT or +869CC, but if "fibrosis-associated" polymorphic sites in other genes could be identified, it may be possible to detect fibrosis prone individuals before radiotherapy with greater certainty. PMID- 12569020 TI - The synthesis of semipermeable membrane microcapsules using in situ cyanoacrylate ester polymerization. AB - A new material for the microencapsulation of biological systems was discovered and characterized with regards to the effects of reaction conditions on product yield. By using poly(cyanoacrylate ester), membrane microcapsules were produced with sufficient strength and porosity to be effective in a process environment for the immobilization and protection of encapsulated material. After synthesizing numerous monomeric cyanoacrylate esters, the n-butyl derivative was discovered to give the best results with regards to microcapsule formation. Microcapsules were prepared by a droplet technique in which an aqueous solution is sprayed into an organic solvent containing the cyanoacrylate ester monomer. By pre-treating the cyanoacrylate ester monomer with an anion exchange resin (Amberlyst A-21, Rohm and Haas, Philadelphia, USA), a significant reduction in the amount of acidic impurities which can adversely affect results was achieved. The use of polyvinylpyrrolidone as a polymerization initiator gave the best results of a variety of polymeric and non-polymeric initiators investigated. Successful encapsulations were achieved using a solvent mixture of 60% (v/v) iso octane/40% trichloroethylene, 0.1% (w/v) polyvinylpyrrolidone initiator, pH 6.5 aqueous encapsulation solution, and 5% (v/v) methylcyanoacrylate/A-21 treated n butylcyanoacrylate (added separately to solvent) made to a 4% (v/v) solution in solvent. Ester monomers were synthesized and used to prepare polymer membranes. PMID- 12569021 TI - A new microencapsulation device for controlled membrane and capsule size distributions. AB - Microbeads and microcapsules, employed for the microencapsulation of bioactive material, should provide sufficient mechanical protection to the encapsulated material, insure an optimal diffusion of desired molecules and, for transplantation-related applications, block the ingress of the imunoagents. Microcapsules are also often required to be smooth, spherical, within narrow size and membrane thickness distributions. In addition, the bioactive material has to be centred within the capsule, whose size should be minimized in relation to the bioactive material in order to optimise the diffusion of active molecules. The production process of such microcapsules should respect the aforementioned constraints and, in addition, be sterile, repeatable, robust, and harmless to the bioactive material while showing a high output. Two prototypes, dedicated to the microencapsulation of bioactive materials are presented. A semi-manual device permits the control of microcapsule properties for small scale (< 10 000 microcapsules), sterile production. An 'automated reaction control' system has also been developed. The features of the former are demonstrated for the repeatable production of 400 microm-microcapsules using the alginate/cellulose sulphate/poly(methylene-co-guanidine) system. The production rate is 500 000 microcapsules/h, with a size distribution within +/-10% and membrane thickness distribution within +/-5 microm. The latter in particular is, to the authors' knowledge, better than can be achieved with currently disclosed technologies, and is due to the precise control of the reaction conditions and time. PMID- 12569022 TI - Polymers for biodegradable medical devices XI. Microencapsulation studies: characterization of hydrocortisone-loaded poly-hydroxybutyrate-hydroxyvalerate microspheres. AB - Hydrocortisone-loaded monolithic microspheres were prepared, using a single emulsion solvent evaporation process, from a range of poly-beta-hydroxybutyrate hydroxyvalerate copolymers in which both molecular weight and hydroxyvalerate content were varied. Many similarities were observed in the effects of process parameters and co-polymer composition on the morphologies of the microspheres, and the morphologies of microcapsules prepared (and previously reported) by a double emulsion process. The yields of the single emulsion process were generally superior to those of the double emulsion process, although these were adversely affected by hydrocortisone incorporation as the molecular weight of the copolymer was reduced. The predominant effect of hydrocortisone incorporation was on polymer morphology, characterized by the appearance of small surface pores; an effect which increased with increasing drug loading. Changes in polymer molecular weight, copolymer composition and process temperature, together with the incorporation of polycaprolactone in the form of a solvent blend, enabled microspheres with a range of morphologies to be produced providing the potential for control of drug release. PMID- 12569023 TI - Oral controlled release formulation of diclofenac sodium by microencapsulation with ethyl cellulose. AB - The aim of this study was to formulate and evaluate microencapsulated controlled release preparations of diclofenac sodium (DFS) using different proportions of ethyl cellulose (EC) as the retardant material to extend the release. The formulated microcapsules were then compressed into tablets to obtain controlled release oral formulations. Phase separation-coacervation technique was employed to prepare microcapsules of DFS using different proportions of EC in cyclohexane. Physical characteristics of microcapsules and their tablets, in vitro release pattern of the designed microcapsules and their tablets prepared from them were studied using USP dissolution apparatus (USP 2000) type 2 (paddle method) in triple distilled water. The prepared microcapsules were white, free flowing and spherical in shape, with the particle size varying from 49.94-52.72 microm. The duration of DFS release from microcapsules was found to be directly proportional to the proportion of EC and, thus, coat thickness. All tablets were of good quality with respect to appearance, drug content uniformity, hardness, weight variation, friability and thickness uniformity. In vitro release study of the tabletted microcapsules in triple distilled water showed a zero order release kinetics and extended release beyond 24 h. A good correlation was obtained between drug release (t(60)) and proportion of EC in the microcapsules. In the case of tabletted microcapsules, very good correlation could be established between t(60), proportion of EC, weight of the tablets and between release rate constant (K) and proportion of EC. All the formulations were highly stable and possessed reproducible release kinetics across the batches. PMID- 12569024 TI - Microsphere entrapped bee-venom phospholipase A2 retains specific IgE binding capacity: a possible use for oral specific immunotherapy. AB - The only specific treatments of allergy are long and exacting desensitization by subcutaneous injections of the allergens. While oral administration of allergens could greatly facilitate these treatments, effective delivery systems are needed to prevent allergen degradation in the gastrointestinal tract and to enable their uptake by Peyer's patches. The potential for bee-venom phospholipase A2 (PLA2) to be used in such oral immunotherapy was tested. For this purpose, PLA2 potential alterations were analysed when encapsulated into poly(D,L-lactide-co-glycolide) microspheres by double emulsion solvent evaporation. It was shown that microencapsulation had only limited effects on the integrity of the entrapped PLA2, which retained its fully specific murine IgE binding capacity. Thus, PLA2 loaded microspheres could represent a potential delivery system for bee venom allergy-specific oral immunotherapy. PMID- 12569025 TI - The effect of solvent removal conditions on performance and release property of protein-loaded microparticles. AB - This study was designed to investigate the influence of solvent removal conditions on the performance and release properties of protein-loaded poly(epsilon-caprolactone) microparticles. The microparticles were prepared by the coacervation method in three different conditions. The effects of vacuum pressure, fabrication temperature and evaporation time on the crystallinity, surface morphology, particle size as well as the yield of microparticles, encapsulation efficiency of BSA and in vitro release property were investigated. There was no significant difference in the size of microparticles prepared by varying the vacuum pressure and temperature. Similar results were obtained for the production yield of microparticles and the loading efficiency of protein in these microparticles. However, accelerating the evaporation rate of solvent significantly reduced the crystallinity of polymer from 54.13 +/- 2.67% down to 44.64 +/- 2.17% (p < 0.05). The release of protein from the resulting microparticles was rapid, within 6 h, after which BSA was continuously and slowly released for up to 7 days. The protein release rate and polymer crystallinity possessed a good correlation (r = -0.951). This result indicated that the higher the crystallinity, the slower the release rate. In other words, change in vacuum pressure and temperature reduced the crystallinity of polymer, which was feasible for protein to release from amorphous domain in microparticles. PMID- 12569026 TI - Characterization of structure and diffusion behaviour of Ca-alginate beads prepared with external or internal calcium sources. AB - Ca-alginate beads were prepared with either external or internal calcium sources. The structures of both beads were investigated with the aid of scanning electron microscopy (SEM) and confocal microscopy. It was shown that the beads with internal calcium source had a looser structure and bigger pore size than those with external calcium source. The attempts to interpret the difference were carried out by determining the Ca content within the beads at various times, which indicated that it was the different gelation mechanisms that caused the difference of structures of both beads. Furthermore, it was also found that the diffusion rate of haemoglobin (Hb) within the beads with an internal calcium source was faster than that of the beads with an external one, which was consistent with the observation of their structures. PMID- 12569027 TI - The effects of infiltration on protein release from multi-phase microspheres fabricated via solvent removal. AB - Multi-phase polymer microspheres for drug encapsulation have been fabricated via solvent removal using poly(L-lactic) acid (PLLA) and poly(fumaric-co-sebacic) anhydride (P(FA:SA)20:80). A process of protein infiltration by the polymer solution was studied to determine the effect on protein release. Additionally, multiple variations of the infiltration process were investigated. The mechanisms involved in the different infiltration methods were looked at to determine if polymer degradation was occurring or if the protein was aggregating as a result of the infiltration process. Multiple drugs were used in these studies: FITC labelled bovine serum albumin (BSA), a model drug and antide (a GnRH antagonist), which is a therapeutic agent. Characterization and comparison of the various microsphere batches was performed via scanning-electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), gel-permeation chromatography (GPC) and differential-scanning calorimetry (DSC). Protein infiltration by the polymer solution was successful in decreasing the initial burst of drug with insignificant differences between the various methods of infiltration. Furthermore, there were minimal differences in polymer degradation due to the different methods of infiltration and there were no significant differences in the degree of crystallinity of the polymers in the various batches fabricated with and without infiltration. PMID- 12569028 TI - Reacetylated chitosan microspheres for controlled delivery of anti-microbial agents to the gastric mucosa. AB - The high aqueous solubility of chitosan restricts the utility of chitosan microspheres for gastric drug delivery. This paper describes the preparation of reacetylated chitosan microspheres with suitable properties for the controlled release of active anti-microbial agents, such as amoxycillin and metronidazole, in the gastric cavity. Two different microencapsulation approaches were developed and optimized in order to encapsulate hydrophilic (amoxycillin) and hydrophobic (metronidazole) compounds efficiently. The reacetylated chitosan microspheres exhibited a controlled water swelling capacity and gelified at acidic pH, resulting in prolonged release of the encapsulated antibiotics. The reacetylation time was found to be a key factor that affects not only drug release, but also encapsulation efficiency and anti-microbial activity of the encapsulated compound. The last two parameters were also dependent on drug solubility in the reacetylating agent. Using short reacetylation time periods, it was possible to efficiently control the release of both hydrophilic and lipophilic antibiotics while maintaining their activity against different bacteria. Consequently, reacetylated chitosan microspheres are promising vehicles for the controlled delivery of anti-microbial agents to the gastric cavity and, hence, for the eradication of Helicobacter pylori, a pathogen strongly associated with gastric ulcers and possibly gastric carcinoma. PMID- 12569030 TI - Literature alerts. PMID- 12569029 TI - Preparation and characterization of drug-loaded polymethacrylate microspheres by an emulsion solvent evaporation method. AB - Microspheres containing the anti-hypertension drug, felodipine, were prepared by the emulsion solvent evaporation method (o/o) using acrylate methacrylate copolymers, Eudragit RL PO and Eudragit RS PO, as wall materials. In order to increase the encapsulation efficiency, a mixed solvent system comprising 1:1 proportions of acetonitrile and dichloromethane was used as a dispersed phase. The morphology of the microspheres was evaluated using a scanning electron microscope, which showed a spherical shape with smooth surface. The mean sphere diameter was between 9.5-13.2 microm and the microencapsulation efficiencies ranged from 51.4-80.4%. The release profiles and encapsulation efficiencies depended strongly on the structure of the polymers used as wall materials. The release rate of the Eudragit RS PO microspheres was much lower than that of Eudragit RL PO microspheres. Whereas Eudragit RL PO microspheres followed the Higuchi rule, Eudragit RS PO microspheres exhibited a triphasic release profile. It is concluded that drug release rate can be controlled by choice of polymer type. PMID- 12569033 TI - Bullying risks of 11-year-old children with specific language impairment (SLI): does school placement matter? AB - In addressing an issue rarely explored in research literature, the prevalence and severity of the risk of being bullied at school was measured in 100 children with specific language impairment (SLI). Participants attended a range of different educational placement types and these were compared for bullying risk. Furthermore, the risk encountered by children with SLI was compared with that of normally developing age-matched peers. Each participant completed a questionnaire and it was found that 36% of participants with SLI considered themselves at risk of being bullied in school compared with only 12% of the normally developing cohort. No statistically significant difference was found between the risk experienced by participants with SLI attending mainstream education and that by participants attending special education placements. Possible explanations for the results are offered and the relevance of the findings in the context of optimizing the educational experience of children with SLI is highlighted. PMID- 12569034 TI - Intergenerational communication: fundamental but under-exploited theory for speech and language therapy with older people. AB - There is a body of research literature already applied in speech and language therapy practice that is concerned with communication between children and adults and the adaptations adults make to facilitate the development of language in children. There is much less and more recent literature concerned with intergenerational communication involving older people and older people in institutional care. This has not yet impacted on speech and language therapy practice, especially in the area of training others. The aims of this paper are (1) to describe some of the main theoretical concepts associated with intergenerational communication, (2) to present the results from a study of the opinions about and experiences of intergenerational communication in which children, community- based older women and professional carers of older people were included and (3) to discuss the implications for speech and language therapy practice. A hypothesis for the study was that views on and attitudes towards communication and ageing would vary among the age groups. Some of the main concepts and models associated with intergenerational communication with older people are reviewed, including the communication predicament and enhancement models and the concept of patronizing communication. A qualitative study of three different age groups of (mainly) women was undertaken using a variety of methods of data elicitation (including written questionnaire and focus group discussion). Themes arising from the data were illuminated using content analysis. Participants' responses demonstrate some current generally positive views across the life-span on what it means to be old and the value of communication with older people. The implications for speech and language therapy practice are outlined, with the main emphasis on the potential use of intergenerational communication theory in developing a new focus for training other staff groups who care for older people and for measures of effectiveness of such in-service training. PMID- 12569035 TI - Evaluation of speech in people with head and neck cancer: a pilot study. AB - BACKGROUND: A paucity of information exists on the effects of cancer on speech, with what little literature exists focussing on the effects of treatment. Baseline assessment of speech in head and neck cancer patients before treatment is important to distinguish accurately the effects of treatment from the effects of the cancer itself. AIMS: This prospective study had four major foci: (1) to determine if speech intelligibility and motor speech functions of patients with head and neck cancer before treatment differed from the normal population; (2) to determine if impairments in speech intelligibility and motor speech functions were specific to the original site of cancer; (3) to determine which motor speech functions were most related to overall speech intelligibility; and (4) to determine whether there was a relationship between self-report of speech intelligibility, motor speech functions and the clinical assessment thereof. This study aimed incidentally to determine whether the Frenchay Dysarthria Assessment is a practicable, valid and reliable tool for assessing speech functions in the head and neck cancer population. METHODS & PROCEDURES: Twenty participants, between 47 and 76 years of age and all newly diagnosed with a cancer of the head and neck, participated in the pilot study. In examining speech, participants completed a self-report questionnaire pertaining to speech intelligibility and motor speech functions and were then assessed clinically using subscales of the Frenchay Dysarthria Assessment before the start of cancer treatment. The motor speech functions examined included: respiratory ability and functions of the lip, soft palate, larynx and tongue. OUTCOMES & RESULTS: Results indicated that participants with head and neck cancer had a greater reduction in both speech intelligibility and in almost all aspects of speech when compared with the normal population. Results further demonstrated that the site of the cancer dictated the type of impairment experienced with respect to speech function. Not all motor speech characteristics correlated with overall speech intelligibility and, lastly, self-report and clinical assessment were significantly correlated with respect to motor speech characteristics, but they differed slightly in the assessment of speech intelligibility. CONCLUSIONS: This pilot study has highlighted the critical importance of collecting baseline measures and reporting functional results according to the cancer site in future research. It has also shown the usefulness of the Frenchay Dysarthria Assessment as a practicable, valid and reliable protocol of motor speech assessment for the head and neck cancer population. PMID- 12569036 TI - Changes in linguapalatal contact patterns during therapy for velar fronting in a 10-year-old with Down's syndrome. AB - BACKGROUND: Articulation errors in the speech of people with Down's syndrome are frequent and often resistant to speech therapy. This preliminary study investigates the use of electropalatography (EPG) to diagnose and treat abnormal articulation patterns associated with velar fronting in a 10-year-old girl. AIMS: The study measured changes in the accuracy and stability of linguapalatal (tongue palate) contact patterns during a 14-week course of visual feedback therapy using EPG. Therapy aimed to resolve a pattern of velar fronting whereby targets /k, g, eta/ had alveolar placement [t, d, n]. METHODS & PROCEDURES: The participant was a girl (P) with Down's syndrome aged 10;11 years. P had a moderate-severe speech disorder, which included velar fronting. Her speech was recorded with EPG on three occasions during a 14-week course of therapy: first, before therapy; second, midway through therapy; and third, after therapy. Three analyses were conducted on the EPG data. The first used an EPG classification scheme that identified accuracy of placement for /t/ and /k/ targets. The second was a centre of gravity measure that detected whether P produced a significant difference between /t/ and /k/ targets. The third was a variability index that quantified the stability of contact patterns. OUTCOMES & RESULTS: The results of the EPG classification showed that before therapy, /t/ and /k/ targets had identical alveolar placement, reflecting the process of velar fronting. The results after therapy showed that 87% of /k/ targets had accurate velar placement. The centre of gravity measure showed no difference in contact patterns for /t/ and /k/ before therapy, but a statistically significant difference at the second and third recordings. The variability index showed stable contact patterns before therapy for /t/ and /k/ targets, but both became highly unstable midway through therapy, with a return to stability at the third recording. We embed a discussion of P's increased articulation instability during therapy in a recent theoretical framework--dynamic systems--that attempts to account for the emergence of new behavioural forms. CONCLUSIONS: These preliminary results suggest that EPG has potential as an effective diagnostic and therapy procedure for articulation errors in people with Down's syndrome. A major issue still to be addressed, however, is the extent to which others will benefit from this approach to intervention. PMID- 12569037 TI - Evaluating UK research in speech and language therapy. AB - BACKGROUND: There has been a steady growth in recent years in British higher degree training in speech and language therapy. But what is the standing of UK research in the subject and its component areas which should underpin and inform such training? How can such research be evaluated? AIMS: The intention was to compare UK publications relevant to speech and language therapy with those of other countries, both quantitatively and qualitatively. We sought then to examine the UK papers in more detail to analyse their sources of funding, their geographical distribution and the ways in which they could appropriately be evaluated. METHODS & PROCEDURES: Papers were selectively retrieved from the Science Citation Index and the Social Sciences Citation Index for 1991-2000 by means of a filter based on journal names and paper title words. They were subsequently checked to remove many false positives. The papers were classified into one of seven subject areas and by their research level (from clinical to basic). Their importance was estimated through their potential impact on other researchers, as determined by the citation score of their journals, by the numbers of citations they actually received and by the subjective esteem in which the various journals were held by UK speech and language researchers. OUTCOMES & RESULTS: World output of speech and language therapy papers has averaged 1000 papers per year during the 1990s, and has grown by half over the period. UK output has been about 12% of the total, compared with 10% in biomedicine, and is published in high impact journals relative to the norm for the field, which is quite a low rate compared with biomedicine overall. Almost half the UK papers had no funding acknowledgements, with the private-non-profit and industrial sectors playing less of a role than in other biomedical areas. Papers in seven subject areas showed substantial differences in their performance on the four criteria selected. CONCLUSIONS: The state of British speech and language research appears to be satisfactory, with an above average output in both quantity and quality. However, it is not attracting funding from some types of sponsors and is not being published in general medical journals where it might have a wider influence on general clinical practice. It is also not clear how best such research can be evaluated, although conventional citation counts may be relevant for some subject areas. PMID- 12569038 TI - Is speech and language therapy meeting the needs of language minorities? The case of deaf people with neurological impairments. AB - The cultural diversity of the UK poses a challenge for speech and language therapists. Work with children from language minorities has been documented, but less so with adults. This raises the question of whether adults from minority communities are gaining access to services. Deaf people who use British Sign Language (BSL) are one language minority. It is known that sign language is vulnerable to neurological damage. Of interest is whether Deaf people with such damage are referred to speech and language therapy (SLT). The aim was to find out how many deaf people were referred to SLT services in the UK over the last 5 years, and reasons for the referrals. We also explored the service offered to Deaf referrals, and whether SLT teams had access to BSL skills, either internally or via interpreters. A brief questionnaire was sent to the managers of all SLT services in the UK (n=264). There was as 60% response rate. Only 34 services received referrals of Deaf patients, with a total of 39 Deaf people seen. This is substantially below the predicted rates. Most referrals were for dysphagia, or dysphagia with communication impairments. In line with this, dysphagia management was the dominant service, although most referrals also received language assessment. Most teams did not have signing staff members and access to interpreters was variable. The results suggest that many Deaf people are not gaining access to SLT after neurological impairment. Those who are referred are unlikely to receive language therapy. The instigation of a national team specializing in BSL impairments is recommended. PMID- 12569039 TI - Conversation analysis as a technique for exploring the dynamics of a mediated interview. AB - The study analysed the dynamics of a mediated medico-legal interview using conversation analysis (CA) as a key methodology. The aim of using CA was to identify both facilitators and inhibitors of a successfully mediated interview, using a detailed microscopic analysis of the dynamics involved. A 45-minute interview with the client's parents, the speech pathologist and the interpreter was tape recorded and analysed according to CA principles. Results revealed several facilitators, including equal and active roles, use of code switching, familiarity between the interviewer and interpreter, and use of repetition. Inhibitors included different agendas, complicated repair trajectories and interruptions. In addition, aspects such as cultural brokerage were identified that could be considered neither as facilitators nor as inhibitors. Each area is discussed in detail using extracts from the transcription. PMID- 12569042 TI - Frame of reference and adaptation to directional bias in a video-controlled reaching task. AB - The present study (N=56) investigated spatio-temporal accuracy of horizontal reaching movements controlled visually through a vertical video monitor. Direct vision of the hand was precluded and the direction of hand trajectory, as perceived on the video screen, was varied by changing the angle of the camera. The orientation of the visual scene displayed on the fronto-parallel plane was thus congruent (0 degrees condition) or non-congruent (directional bias of 15 degrees, 30 degrees or 45 degrees counterclockwise) according to the horizontal working space. The goal of this study was to determine whether local learning of a directional bias can be transferred to other locations in the working space, but taking into account the magnitude of the directional bias (15 degrees, 30 degrees or 45 degrees ), and the position of the successive objectives (targets at different distances (TDD) or different azimuths (TDA)). Analysis of the spatial accuracy of pointing movements showed that when introducing a directional bias, terminal angular error was linearly related to the amount of angular perturbation (around 30%). Seven trials were, on average, necessary to eliminate this terminal error, whatever the magnitude of the directional bias and the position of the successive targets. When changing the location of the spatial objective, transfer of adaptation was achieved in the TDD condition but remained partial in the TDA condition. Furthermore, initial orientation of the trajectory suggested that some participants used a hand-centred frame of reference whereas others used an external one to specify movement vector. The adaptation process differed as a function of the frame of reference used, but only in the TDA condition. Adaptation for participants using a hand-centred frame of reference was more concerned with changes in the shape of the trajectory, whereas participants using an external frame of reference adapted their movement by up dating the initial direction of hand trajectory. As a whole, these findings suggest that the processes involved in remote visual control of hand movement are complex with the result that tasks requiring video-controlled manipulation like video-controlled surgery require specific spatial abilities in actors and consequential plasticity of their visuo-motor system, in particular concerning the selection of the frame of reference for action. PMID- 12569043 TI - A maritime navigation display that provides visual feedback to improve conning officers' ship-handling during low visibility environments. AB - Navigating a ship at night in close proximity to another vessel is a dangerous task. To improve conning officers' night time ship-handling performance, a visual navigation display is proposed that is mounted on the stern of the aircraft carrier that will help shipboard conning officers manoeuvre in a battle group formation. To test the effectiveness of the visual navigation display, the Nimitz class aircraft and plane guard vessel, a Ticonderoga-class cruiser, were modelled in a virtual environment. A navigation display condition had significantly fewer navigational positional errors than a non-navigation display condition. The navigation display provided immediate feedback as to whether the aircraft carrier had changed bearing or speed, thus enabling the operator to initiate the appropriate input to maintain station astern of the carrier. Actual or potential applications of this research include the deployment of a maritime navigation display to assist conning officers' ship handling. PMID- 12569044 TI - Grasping an augmented object to analyse manipulative force control. AB - Augmented reality allows changes to be made to the visual perception of object size even while the tangible components remain completely unaltered. It was, therefore, utilized in a study whose results are being reported here to provide the proper environment required to thoroughly observe the exact effect that visual change to object size had on programming fingertip forces when objects were lifted with a precision grip. Twenty-one participants performed repeated lifts of an identical grip apparatus to a height of 20 mm, maintained each lift for 8 seconds, and then replaced the grip apparatus on the table. While all other factors of the grip apparatus remained unchanged, visual appearance was altered graphically in a 3-D augmented environment. The grip apparatus measured grip and load forces independently. Grip and load forces demonstrated significant rates of increase as well as peak forces as the size of graphical images increased; an aspect that occurred in spite of the fact that extraneous haptic information remained constant throughout the trials. By indicating a human tendency to rely - even unconsciously - on visual input to program the forces in the initial lifting phase, this finding provides further confirmation of previous research findings obtained in the physical environment; including the possibility of extraneous haptic effects (Gordon et al. 1991a, Mon-Williams and Murray 2000, Kawai et al. 2000). The present results also suggest that existing knowledge concerning human manipulation tasks in the physical world may be applied to an augmented environment where the physical objects are enhanced by computer generated visual components. PMID- 12569045 TI - Perceptual and signal detection factors in radiography. AB - Diagnostic imaging systems (e.g. X-rays and ultrasound) play an important role in medicine. Owing to increasing workloads now carried by clinical radiologists, the interpretation of medical images is increasingly being passed to radiographers. This is especially so in those areas of diagnoses where examinations are performed on large numbers of patients but when the frequency of the disorder is very low (e.g. mass mammography screening). This study explored the accuracy of the radiographic interpretation of pulmonary nodules by clinical radiologists and student radiographers, and their correlations with tests of field-dependency, visual search and spatial reasoning. Overall performance in lung nodule detection gave a mean ROC (Az)=0.851 for student radiographers, while radiologists achieved 0.871. No correlation was found between diagnostic performance and either fielddependency or visual search. A significant negative correlation was found between diagnostic performance and spatial reasoning (p < 0.05). PMID- 12569046 TI - The metabolic cost of operating a bicycle generator light. AB - The purpose of the study was to determine the metabolic cost of operating a bicycle generator (dynamo) light. Nineteen subjects (12 males, 7 females) volunteered to participate in the study. All tests were conducted using a multigeared road bicycle mounted on a Velodyne computer-controlled, electromagnetically-braked bicycle training simulator. A tyre sidewall-driven 6V 3W bicycle generator was attached to the bicycle frame. Tyre pressure was maintained at 6.12 atm. Gears were prescribed to produce simulated riding speeds of approximately 13 and 21 km h(-1) at 62 rpm. Subjects rode four, 5-min stages during the test session to evaluate riding under conditions of generator OFF or ON at each speed. Oxygen consumption, heart rate, and rating of perceived exertion (RPE) were recorded at the end of each minute. Paired samples t tests revealed significant differences between generator OFF and ON conditions for both the metabolic cost of cycling and heart rate at each of the speeds tested. There were no significant differences found between conditions for RPE. Bicycling with the generator ON increased oxygen consumption by 8.4% at 13 km h(-1) and 9.6% at 21 km h(-1). PMID- 12569048 TI - Creative (dis)agreement in ergonomics. PMID- 12569049 TI - Subjective rating scales: science or art? AB - Subjective rating scales are widely used in almost every aspect of ergonomics research and practice for the assessment of workload, fatigue, usability, annoyance and comfort, and lesser known qualities such as urgency and presence, but are they truly scientific? This paper raises some of the key issues as a basis for debate. First, it is argued that all empirical observations, including those conventionally labelled as 'objective', are unavoidably subjective. Shared meaning between observers, or intersubjectivity, is the key criterion of scientific probity. The practical steps that can be taken to increase intersubjective agreement are discussed and the well-known sources of error and bias in human judgement reviewed. The role of conscious experience as a mechanism for appraising the environment and guiding behaviour has important implications for the interpretation of subjective reports. The view that psychometric measures do not conform to the requirements of truly 'scientific' measurement is discussed. Human judgement of subjective attributes is essentially ordinal and, unlike physical measures, can be matched to interval scales only with difficulty, but ordinal measures can be used successfully both to develop and test substantive theories using multivariate statistical techniques. Constructs such as fatigue are best understood as latent or inferred variables defined by a set of manifest or directly observed indicator variables. Both construct validity and predictive validity are viewed from this perspective and this helps to clarify several problems including the dissociation between measures of different aspects of a given construct, the question of whether physical (e.g. physiological) measures should be preferred to subjective measures and whether a single measure of constructs which are essentially multidimensional having both subjective and physical components is desirable. Finally, the fitness of subjective ratings to different purposes within the broad field of ergonomics research is discussed. For testing of competing hypotheses concerning the mechanisms underlying human performance, precise quantitative predictions are rarely needed. The same is frequently true of comparative evaluation of competing designs. In setting design standards, however, something approaching the level of measurement required for precise quantitative prediction is required, but this is difficult to achieve in practice. Although it may be possible to establish standards within restricted contexts, general standards for broadly conceived constructs such as workload are impractical owing to the requirement for representative sampling of tasks, work environments and personnel. PMID- 12569050 TI - Measurement and the practising ergonomist. PMID- 12569051 TI - Sans subjectivity - ergonomics is engineering. PMID- 12569052 TI - Is ergonomics empirical? PMID- 12569053 TI - Subjective measures: not perfect but what is? PMID- 12569054 TI - Transactional perspectives on subjective rating scales. PMID- 12569055 TI - Use of subjective rating scores in ergonomics research and practice. PMID- 12569056 TI - Do ratings measure latent attributes? PMID- 12569057 TI - Subjective rating scales: scientific measures of perceived urgency? PMID- 12569058 TI - Measurements serve a purpose: a note on a possible use of natural measurements. PMID- 12569059 TI - It's all relative: defining mental workload in the light of Annett's paper. PMID- 12569060 TI - Subjective evaluation of usability. PMID- 12569061 TI - On the assessment of (mental) workload and other subjective qualifications. PMID- 12569062 TI - Measurement in virtual environments: another dimension to the objectivity/subjectivity debate. PMID- 12569063 TI - Can design supportive research be scientific? PMID- 12569065 TI - RSD931, a novel anti-tussive agent acting on airway sensory nerves. AB - 1 The anti-tussive effects, of the local anaesthetic, lidocaine and carcainium chloride (RSD931) have been investigated in guinea-pigs and rabbits. 2 Pre treatment of guinea-pigs with aerosols of lidocaine or RSD931 at 0.1, 1.0 and 10 mg ml(-1) reduced the number of citric acid-induced coughs by 9.3, 32.6 and 40.9% (P>0.05) for lidocaine and by 25.3% (P>0.05), 40.4% (P>0.05) and 97.6% (P<0.01) for RSD931, respectively and increased the latency to onset of cough at 10.0 mg ml(-1) only. In addition, RSD931 at 10 mg ml(-1) reduced citric acid-evoked cough responses in rabbits (with prior exposure to ozone at 3 p.p.m. for 1 h) from 22.1+/-5.1 to 2.7+/-0.9 coughs (P<0.01). 3 Acute pre-treatment of guinea-pigs with aerosols of lidocaine or RSD931 at 10.0 and 30.0 mg ml(-1) reduced the number of capsaicin-evoked coughs by 42.2 and 10.3% (P>0.05) (lidocaine) and by 25% (P>0.05) and 76.9% (P<0.01) (RSD931), respectively. Lidocaine had little effect on the latency of cough onset at either 10.0 or 30.0 mg ml(-1), however, RSD at 30.0 mg ml(-1) significantly (P<0.05) prolonged the latency of cough onset. 4 RSD931 (10.0 mg ml(-1)) significantly (P<0.05-<0.01) reduced the spontaneous and histamine-evoked discharges in Adelta-fibres originating from airway, rapidly adapting stretch receptors (RARs) without affecting histamine evoked bronchoconstriction. Lidocaine at 10.0 mg ml(-1) also significantly (P<0.05) inhibited the spontaneous and histamine-induced discharges of RARs without affecting histamine-evoked bronchoconstriction. 5 Aerosols of RSD931 (10.0 mg ml(-1)) caused a transient, but significant (P<0.05), activation of pulmonary C-fibre endings 2.5 min after administration started. RSD931 had no significant (P>0.05) effects on discharges in bronchial C-fibres originating from bronchial C-fibre endings, capsaicin-evoked discharges of either pulmonary or bronchial C-fibre endings or on capsaicin-evoked bronchoconstriction. In contrast, lidocaine (10.0 mg ml(-1)) significantly (P<0.05) inhibited spontaneous and capsaicin-induced discharges in both pulmonary and bronchial C-fibres respectively. Lidocaine also significantly (P<0.05) reduced capsaicin-evoked bronchoconstriction. 6 These studies suggest that the anti-tussive actions of RSD931 are mediated via inhibition of discharges in Adelta-fibres originating from airway RARs. The mechanism of action of RSD931 is distinct from that of the local anaesthetic lidocaine and RSD931 may represent a novel class of anti tussive agent. PMID- 12569066 TI - The effect of chelerythrine on depolarization-induced force responses in skinned fast skeletal muscle fibres of the rat. AB - 1 We examined the effect of the protein kinase C (PKC) inhibitor chelerythrine on depolarization-induced force responses (DIFRs) and sarcoplasmic reticulum (SR) function in single, mechanically skinned skeletal muscle fibres of the rat. 2 In this study, the DIFRs in the skinned fibres normally underwent an irreversible loss of excitation-contraction coupling (ECC) after 10-15 responses. Chelerythrine (12 micro M) was shown to restore ECC in these fibres. Restored force responses were similar in peak (control 50.8+/-6.4%, chelerythrine 56.9+/ 12.4% of maximum force, P=0.42, n=21), but significantly broadened compared to initial control responses (full-width at half maximum, control; 3.7+/-0.3 s, chelerythrine; 13.3+/-1.1 s, P<0.001). Early exposure to chelerythrine prevented run-down of DIFRs. Chelerythrine also induced spontaneous force responses in some fibres. 3 The PKC inhibitors calphostin C and staurosporine did not restore ECC, and the PKC activator phorbol 12-myristate 13-acetate did not promote loss of ECC in the skinned fibres. 4 Chelerythrine significantly increased SR Ca(2+) loading by 8.4+/-1.7% (P=0.02, n=9) and SR Ca(2+) release by at least 14.1+/-2.7% (P=0.004, n=11) in the skinned fibres. 5 Chelerythrine had no significant effect on maximum force production or the [Ca(2+)] producing half maximal activation of the myofilaments. However, chelerythrine did have a small effect on the slope of the force-Ca(2+) relationship (P=0.02, n=10). 6 Chelerythrine reverses the use dependent loss of excitation-contraction coupling in skinend skeletal muscle fibres by a PKC independent pathway. Chelerythrine may be an important pharmacological probe for examining the mechanisms of contraction-induced muscle injury. PMID- 12569067 TI - A unique phenotype of 5-HT2C, agonist-induced GTPgamma35S binding, transferable to 5-HT2A and 5-HT2B, upon swapping intracellular regions. AB - 1 The human 5-HT(2C) receptor, when expressed heterologously in various mammalian cell lines (HEK293, SH-EP and NIH-3T3) at various receptor densities (6 to 45 pmol mg(-1) protein), mediates robust agonist-induced GTPgamma(35)S binding from coupling to G(i) subtypes of G proteins, in addition to G(q/11). Such a phenotype, however, was not seen with the human 5-HT(2A) and 5-HT(2B) receptors, indicating their common pathway with 5-HT(2C) limited to G(q/11), not including G(i). 2 Because intracellular regions are largely responsible for signalling pathways, we prepared the chimeras of the 5-HT(2A) and 5-HT(2B) receptors where the second and third intracellular loops, and the C-terminal region were replaced with the 5-HT(2C) counterparts. 3 The chimeras showed robust agonist-induced GTPgamma(35)S binding. Relative intrinsic efficacies of agonists from the GTPgamma(35)S binding were nearly identical to the reported values for their parent receptors as measured with Ca(2+) or [(3)H]-inositol phosphate accumulation. Also the chimeras displayed the same ligand-binding properties as the parent receptors. 4 We conclude that the phenotype of agonist-induced GTPgamma(35)S binding is unique to 5-HT(2C) among the 5-HT(2) receptor family, and is transferable to 5-HT(2A) and 5-HT(2B), upon swapping intracellular sequences, without altering their receptor pharmacology. PMID- 12569068 TI - Sphingosylphosphorylcholine, a naturally occurring lipid mediator, inhibits human platelet function. AB - 1 The lysophospholipids, lysophosphatidic acid and sphingosine 1-phosphate, have been reported to activate platelets. Here we examined effects of the naturally occurring related sphingosylphosphorylcholine (SPC) on human platelet function. 2 Platelet activation was determined as aggregation, elevation of intracellular Ca(2+) concentrations, surface expression of P-selectin, GP 53, and GP IIb/IIIa neoepitope PAC-1, and of fibrinogen binding to the platelet surface. 3 Platelets were activated by ADP (5 and 20 micro M), the thrombin receptor-activating peptide TRAP-6 (5 and 20 micro M), the thromboxane A(2) mimetic U-46619 (1 micro M) and collagen (20 and 50 micro g ml(-1)) but not by SPC (up to 20 micro M). 4 SPC concentration-dependently (IC(50) approximately 1-10 micro M) inhibited activation of washed human platelets in response to all of the above agonists, with almost complete inhibition occurring at 20 micro M SPC. 5 The SPC stereoisomers, D-erythro SPC and L-threo SPC, exhibited similar concentration response curves in inhibiting 20 micro M ADP-induced platelet aggregation, suggesting that SPC did not act via specific lysophospholipid receptors. 6 Although SPC slightly activated platelet protein kinase A (as assessed by VASP phosphorylation), this effect could not explain the marked platelet inhibition. Possible protein kinase C inhibition also did not explain the inhibition of platelet activation by SPC. On the other hand, SPC suppressed agonist-induced Ca(2+) mobilization and phospholipase C stimulation. 7 These results indicate that the lysophospholipid SPC is an effective inhibitor of human platelet activation, apparently primarily by uncoupling agonist-activated receptors from their effectors. PMID- 12569069 TI - Characterization of human 5-HT4(d) receptor desensitization in CHO cells. AB - 1 Serotonin 5-HT(4) receptor isoforms differ in their C-terminal tail and yet little is known about their regulation. In this study, we investigated the desensitization of two human 5-HT(4) receptors stably expressed in CHO cells, with a special emphasis on the h5-HT(4(d)) isoform. 2 Exposure of h5-HT(4(d)) and h5-HT(4(e)) receptors to 1 micro M 5-HT induced a rapid desensitization of the adenylyl cyclase response. The h5-HT(4(d)) receptor desensitized with a faster rate (t(1/2)<5 min) than the h5-HT(4(e)) receptor (t(1/2)=15 min) and after 10 min 5-HT treatment cAMP production was reduced by approximately 70%. 3 5-HT induced h5-HT(4(d)) receptor desensitization was mimicked by 8-Bromo-cAMP, a cAMP analogue, and was inhibited by [n-[2-(p-bromocinnamylamino)ethyl]-5 isoquinolinesulphonamide, 2HCl] (H-89), an inhibitor of cAMP-dependent protein kinase (PKA). Inhibitors of endocytosis (sucrose, 0.45 M and concanavaline A, 0.25 mg ml(-1)) partially reversed the h5-HT(4(d)) receptor desensitization process. 4 Given the prominent role of PKA in agonist-induced desensitization, we mutated the four putative PKA phosphorylation sites present in the third intracellular loop (Ser242, Thr253, Thr255) and the C terminal tail (Ser338) of the h5-HT(4(d)) receptor. Surprisingly, mutated receptors in which either one or all four putative phosphorylation sites were substituted to alanine did not impair receptor desensitization suggesting that PKA might act on nonconsensus sites. 5 Altogether, our data demonstrate that the C-terminal tail of h5-HT(4) receptors may influence the rate of agonist-induced desensitization and we provide evidence for a major role of PKA in h5-HT(4(d)) receptor desensitization. PMID- 12569070 TI - Contribution of Na+ -Ca2+ exchanger to pinacidil-induced relaxation in the rat mesenteric artery. AB - 1 Pinacidil relaxes blood vessels through opening the K(ATP) channels with a resultant membrane hyperpolarization and inhibition of Ca(2+) influx. The aim of this study was to examine the mechanisms thereby pinacidil induces K(+) channel independent relaxation in isolated endothelium-denuded rat mesenteric artery. 2 Pinacidil-induced relaxation was inhibited by glibenclamide (1-10 micro M) in phenylephrine-preconstricted rings, but was unaffected by glibenclamide after inhibition of K(+) channels and VGCCs. Pinacidil-induced K(+) channel-independent relaxation remained unchanged after treatment with cyclopiazonic acid (10 micro M), thapsigargin (1 micro M), ouabain (100 micro M), propranolol (10 micro M), Rp cAMPS triethylamine (30 micro M), L-NNA (100 micro M), or ODQ (10 micro M). 3 Pinacidil induced more relaxant effect in the presence of nifedipine than in the presence of 60 mM K(+) plus nifedipine. Pretreatment with Na(+)-Ca(2+) exchanger inhibitors, nickel (30-300 micro M) or benzamil (20 micro M) attenuated pinacidil induced relaxation in normal or in nifedipine-containing solution. Pinacidil (1 micro M) produced less relaxant effect with decreasing extracellular Na(+) concentration. Na(+)-free condition abolished the inhibitory effect of benzamil. Both nickel and benzamil inhibited pinacidil-induced relaxation in the presence of glibenclamide (10 micro M). Nickel (300 micro M) did not affect the relaxant response to sodium nitroprusside. 4 Pinacidil relaxed the rings preconstricted by active phorbol and U46619 with similar potency. 5 The present results indicate that stimulation of the forward mode Na(+)-Ca(2+) exchange pathway is in part responsible for pinacidil-induced K(+) channel-independent vasorelaxation. Pinacidil also induces K(+) channel-dependent but VGCCs-independent relaxation. The PKC-mediated cellular pathway may be a target site for pinacidil only in higher concentrations. PMID- 12569071 TI - Interleukin-1beta inhibits ATP-induced protein kinase B activation in renal mesangial cells by two different mechanisms: the involvement of nitric oxide and ceramide. AB - 1 Extracellular nucleotides, like ATP and UTP, have been shown to activate the protein kinase B (PKB) pathway in mesangial cells. In this study we report that the pro-inflammatory cytokine interleukin-1beta (IL-1beta) inhibits ATP-induced PKB activation. 2 Pretreatment of mesangial cells with IL-1beta leads to a time dependent decrease of ATP-induced PKB phosphorylation. Maximal inhibition is seen after 6 h of pretreatment. Incubating cells with IL-1beta in the presence of the NO synthase inhibitor, N-monomethyl-L-arginine (L-NMMA), reversed the IL-1beta inhibition of PKB activity. A similar decrease in ATP-evoked PKB activation is obtained when cells were pretreated with the nitric oxide (NO) donor, (Z)-1-[2 Aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (Deta-NO), but not with the cell-permeable cGMP analogue, 8-bromo-cGMP. 3 The NO- and IL-1beta mediated delayed inhibition of PKB activity is completely reversed by the phosphatase inhibitor calyculin A, but not by ocadaic acid, suggesting that NO upregulates a protein phosphatase activity, which most probably belongs to the group of protein phosphatases type 1. 4 In addition, IL-1beta also triggers a short-term and transient inhibitory effect on ATP-induced PKB activation which is maximal after 2-5 min of pre-incubation with IL-1beta. This effect occurs independently of NO generation, because no NO synthase is expressed at that time, and consequently, L-NMMA does not reverse the effect. Rather an involvement of the sphingolipid ceramide is likely, since IL-1beta triggers rapid ceramide formation and incubation of cells with the cell-permeable C6-ceramide blocked ATP induced PKB phosphorylation. 5 In summary, our data show that IL-1beta exerts both short-term and long-term inhibitory effects on ATP-stimulated PKB activation, the short-term effect probably involves ceramide formation, whereas the long-term effect is due to inducible NO synthase induction and subsequent NO formation. These results reveal a further facet in the mechanisms of ceramide- and NO-induced cell death, i.e. by turning off the survival signal elicited by PKB. PMID- 12569072 TI - Heterogeneity of presynaptic muscarinic receptors mediating inhibition of sympathetic transmitter release: a study with M2- and M4-receptor-deficient mice. AB - 1 Presynaptic muscarinic receptors modulate sympathetic transmitter release. The goal of the present study was to identify the muscarinic receptor subtype(s) mediating inhibition of sympathetic transmitter release in mouse atria, urinary bladder and vas deferens. To address this question, electrically evoked noradrenaline release was assessed using tissue preparations from NMRI, M(2)- and M(4)-knockout, and the corresponding M(2)- and M(4)-wildtype mice, after preincubation with (3)H-noradrenaline. 2 The muscarinic agonist carbachol decreased evoked tritium overflow (20 pulses/50 Hz) in each tissue and strain investigated. After deletion of the M(2)-receptor the maximal inhibition by carbachol was significantly reduced (by 41-72%), but not abolished, in all tissues. After deletion of the M(4)-receptor a moderate and significant reduction of the maximal inhibition by carbachol (by 28%) was observed only in the vas deferens. 3 Experiments with the muscarinic antagonists methoctramine and pirenzepine confirmed that the presynaptic muscarinic receptors were predominantly M(2) in atria and bladder and probably a mixture of M(2) and M(4) in the vas deferens. 4 Experiments in the urinary bladder with the cholinesterase inhibitor physostigmine and the muscarinic antagonist ipratropium demonstrated that endogenously released acetylcholine predominantly acted through M(2) receptors to inhibit noradrenaline release. However, the results do not exclude a minor contribution of M(4)-receptors to this endogenous inhibition. 5 In conclusion, our results clearly indicate that the release-inhibiting muscarinic receptors on postganglionic sympathetic axons in mouse atria, bladder and vas deferens represent mixtures of M(2)- and non-M(2)-receptors. The non-M(2) receptors remain unknown in atria and the bladder, and may represent primarily M(4)-receptors in the vas deferens. These results reveal an unexpected heterogeneity among the muscarinic receptors mediating inhibition of noradrenaline release. PMID- 12569073 TI - Regulated and constitutive activation of specific signalling pathways by the human S1P5 receptor. AB - 1 We tested the hypothesis, whether G Protein-coupled receptors (GPCRs) may differentially regulate specific signalling pathways by constitutive and agonist induced activation using the human sphingosine 1-phosphate receptor S1P(5) as a model. 2 S1P(5) receptor-expressing HEK293 cells exhibited a high degree of basal activity for both inhibition of adenylyl cyclase and extracellular signal regulated kinase (ERK) when cultured in serum, which contains high levels of sphingosine 1-phosphate (S1P). However, basal activity was independent of the presence of S1P: (i) constitutive activity remained when cells were cultured in delipidated serum, (ii) addition of S1P to delipidated serum did not increase basal S1P(5) receptor signalling. 3 Conversely, constitutive inhibition of forskolin-stimulated adenylyl cyclase was further enhanced by S1P in S1P(5) HEK293 cells. 4 Transfection of several mammalian cell lines (CHO-K1, HEK293, NIH 3T3, RH7777) with the S1P(5) receptor induced cell rounding, which was more pronounced in the presence of S1P-containing serum. Rounded cell morphology did not correlate with apoptotic cell death, but led to detachment of cells. 5 Cell surface ELISA assays showed that a fraction of plasma membrane S1P(5) receptors were dose-dependently internalized with S1P. 6 These data reveal that intrinsic inhibition of unstimulated adenylyl cyclase or ERK activity by the S1P(5) receptor is insensitive to ligand modulation. Conversely, effects on forskolin stimulated adenylyl cyclase, cell morphology and internalization can be further augmented with S1P. Our results suggest that different signal transduction pathways are not equally activated through constitutively active GPCRs with promiscuous signalling characteristics. PMID- 12569074 TI - Attenuation of acute and chronic effects of morphine by the imidazoline receptor ligand 2-(2-benzofuranyl)-2-imidazoline in rat locus coeruleus neurons. AB - 1 The aim of this study was to determine if 2-(2-benzofuranyl)-2-imidazoline (2 BFI) interacts with the opioid system in the rat locus coeruleus, using single unit extracellular recordings. 2 In morphine-dependent rats, acute administration of the selective imidazoline receptor ligands 2-BFI (10 and 40 mg kg(-1), i.p. and 100 micro g, i.c.v.) or valldemossine (10 mg kg(-1), i.p.) did not modify the naloxone-induced hyperactivity of locus coeruleus neurons compared with that observed in the morphine-dependent control group. 3 After chronic administration of 2-BFI (10 mg kg(-1), i.p., three times daily, for 5 days) and morphine, naloxone-induced hyperactivity and tolerance to morphine were attenuated. This effect was not observed when a lower dose of 2-BFI (1 mg kg(-1), i.p.) or valldemossine (10 mg kg(-1), i.p.) were used. 4 Acute administration of 2-BFI (10 and 40 mg kg(-1), i.p. and 100 micro g, i.c.v.) but not valldemossine (40 mg kg( 1), i.p.) diminished the potency of morphine to inhibit locus coeruleus neuron activity in vivo (ED(50) values increased by 2.3, 2.9; and 3.1 fold respectively). Similarly, the potency of Met(5)-enkephalin to inhibit locus coeruleus neurons was decreased when 2-BFI (100 micro M) was applied to rat brain slices (EC(50) increased by 5.6; P<0.05). 5 The present data demonstrate that there is an interaction between 2-BFI and the opioid system in the locus coeruleus. This interaction leads to an attenuation of both the hyperactivity of locus coeruleus neurons during opiate withdrawal and the development of tolerance to morphine when 2-BFI is chronically administered. These results suggest that imidazoline drugs may prove to be useful agents for the management of opioid dependence and tolerance. PMID- 12569076 TI - Desensitization of beta2-adrenoceptor-mediated responses by short-acting beta2 adrenoceptor agonists in human lung mast cells. AB - 1 The principal aim of the present study was to determine whether long-term treatment of human lung mast cells (HLMC) with the clinically-relevant beta(2) adrenoceptor agonists, salbutamol and terbutaline, leads to desensitization of beta(2)-adrenoceptor-mediated responses in these cells. 2 The non-selective beta adrenoceptor agonist, isoprenaline, and the selective beta(2)-adrenoceptor agonists, salbutamol and terbutaline, inhibited the IgE-mediated release of histamine from HLMC. Salbutamol (pD(2); 7.7+/-0.3) and terbutaline (pD(2); 7.3+/ 0.2) were roughly equipotent as inhibitors of histamine release although both agonists were less potent than isoprenaline (pD(2); 8.6+/-0.2). 3 Isoprenaline (10(-5) M), salbutamol (10(-5) M) and terbutaline (10(-5) M) enhanced total cell cAMP levels in HLMC over basal by 361+/-90, 150+/-38 and 165+/-35%, respectively. 4 Long-term exposure (24 h) of HLMC to either salbutamol (10(-7) M) or terbutaline (10(-7) M) led to a subsequent reduction in the effectiveness of salbutamol and terbutaline (both 10(-9)-10(-4) M) to inhibit histamine release. However, salbutamol was significantly (P<0.05) more effective than terbutaline at promoting the functional desensitization. 5 Radioligand binding studies, using iodinated cyanopindolol, were performed to determine beta(2)-adrenoceptor density in cell membranes after pretreatment (24 h) of cells with either salbutamol (10( 6) M) or terbutaline (10(-6) M). Both agonists reduced beta(2)-adrenoceptor density in membranes to about the same extent (approximately 25% reduction) but these changes in receptor density were not statistically significant (P>0.05). 6 These data indicate that long-term exposure of mast cells to salbutamol causes greater levels of desensitization to beta(2)-adrenoceptor-mediated responses in HLMC than terbutaline. These findings may have wider clinical significance in the context of asthma treatment as compromised mast cell inhibition could result following long-term exposure of mast cells to short-acting bronchodilators. PMID- 12569075 TI - The anti-inflammatory actions of methotrexate are critically dependent upon the production of reactive oxygen species. AB - 1 The mechanism of action by which methotrexate (MTX) exerts its anti inflammatory and immunosuppressive effects remains unclear. The aim of this study is to investigate the hypothesis that MTX exerts these effects via the production of reactive oxygen species (ROS). 2 Addition of MTX (100 nM-10 micro M) to U937 monocytes induced a time and dose dependent increase in cytosolic peroxide [peroxide](cyt) from 6-16 h. MTX also caused corresponding monocyte growth arrest, which was inhibited (P<0.05) by pre-treatment with N-acetylcysteine (NAC; 10 mM) or glutathione (GSH; 10 mM). In contrast, MTX induction of [peroxide](cyt) in Jurkat T cells was more rapid (4 h; P<0.05), but was associated with significant apoptosis at 16 h at all doses tested (P<0.05) and was significantly inhibited by NAC or GSH (P<0.05). 3 MTX treatment of monocytes (10 nM-10 micro M) for 16 h significantly reduced total GSH levels (P<0.05) independently of dose (P>0.05). However, in T-cells, GSH levels were significantly elevated following 30 nM MTX treatment (P<0.05) but reduced by doses exceeding 1 micro M compared to controls (P<0.05). 4 MTX treatment significantly reduced monocyte adhesion to 5 h and 24 h LPS (1 micro g ml(-1)) activated human umbilical vein endothelial cells (HUVEC; P<0.05) but not to resting HUVEC. Pre-treatment with GSH prevented MTX induced reduction in adhesion. 5 In conclusion, ROS generation by MTX is important for cytostasis in monocytes and cytotoxicity T-cells. Furthermore, MTX caused a reduction in monocyte adhesion to endothelial cells, where the mechanism of MTX action requires the production of ROS. Therefore its clinical efficacy can be attributed to multiple targets. PMID- 12569077 TI - The preferential beta3-adrenoceptor agonist BRL 37344 increases force via beta1 /beta2-adrenoceptors and induces endothelial nitric oxide synthase via beta3 adrenoceptors in human atrial myocardium. AB - 1 The present study investigated the effects of the preferential beta(3)-AR agonist BRL 37344 (BRL) on force of contraction (FOC), Ca(2+)-transient and eNOS activity in human right atrial myocardium. 2 BRL concentration-dependently caused an increase in FOC that was paralleled by an increase in Ca(2+)-transient and a shortening of time to half peak relaxation (T0.5T). These effects were abolished in the presence of propranolol (0.3 micro M). 3 BRL acted as a competitive antagonist towards isoprenaline and in binding experiments it was shown to have a distinct affinity towards beta(1/2)-AR. 4 In immunohistochemical experiments BRL (10 micro M) increased detection of activated eNOS. This effect remained constant in the presence of propranolol (0.3 micro M). 5 BRL increased directly detected NO in DAF-staining experiments. This increase was significantly smaller in the presence of the NO-inhibitor L-NAME. 6 The inotropic effects of BRL were not changed in the presence of L-NMA. 7 These results suggest that the inotropic effects of BRL in human atrium are mediated via beta(1/2)-AR, whereas the increase of atrial eNOS-activity is due to beta(3)- adrenergic stimulation. This increase in eNOS-activity did not influence atrial myocardial contractility. In conclusion, this study shows that beta(3)-adrenergic stimulation is present in human atrium, but may not be functionally as significant as in the left ventricle. PMID- 12569078 TI - Incidence of serious side effects from first-line antituberculosis drugs among patients treated for active tuberculosis. AB - Major adverse reactions to antituberculosis drugs can cause significant morbidity, and compromise treatment regimens for tuberculosis (TB). Among patients treated for active TB we estimated the incidence, and risk factors, of major side effects from first-line anti-TB drugs. Side effects, resulting in modification or discontinuation of therapy, or hospitalization, were attributed on the basis of resolution after withdrawal, and/or recurrence with rechallenge. Among 430 patients treated between 1990 and 1999, the incidence of all major adverse effects was 1.48 per 100 person-months of exposure (95% confidence interval [95% CI], 1.31 to 1.61) for pyrazinamide, compared with 0.49 (95% CI, 0.42 to 0.55) for isoniazid, 0.43 (95% CI, 0.37 to 0.49) for rifampin, and 0.07 (95% CI, 0.04 to 0.10) for ethambutol. Occurrence of any major side effect was associated with female sex (adjusted hazard ratio, 2.5; 95% CI, 1.3 to 4.7), age over 60 years (adjusted hazard ratio, 2.9; 95% CI, 1.3 to 6.3), birthplace in Asia (adjusted hazard ratio, 2.5; 95% CI, 1.3 to 5.0), and human immunodeficiency virus-positive status (adjusted hazard ratio, 3.8; 95% CI, 1.05 to 13.4). Pyrazinamide-associated adverse events were associated with age over 60 years (adjusted hazard ratio, 2.6; 95% CI, 1.01 to 6.6) and birthplace in Asia (adjusted hazard ratio, 3.4; 95% CI, 1.4 to 8.3), whereas rifampin-associated adverse events were associated with age over 60 years (adjusted hazard ratio, 3.9; 95% CI, 1.02 to 14.9) and human immunodeficiency virus-positive status (adjusted hazard ratio, 8.0; 95% CI, 1.5 to 43). The incidence of pyrazinamide induced hepatotoxicity and rash during treatment for active TB was substantially higher than with the other first-line anti-TB drugs, and higher than previously recognized. PMID- 12569079 TI - A small molecule very late antigen-4 antagonist can inhibit ovalbumin-induced lung inflammation. AB - A nonpeptidyl small molecule antagonist, compound A, to nonactivated very late antigen-4 (VLA4) was examined in lung inflammation induced by a single dose of ovalbumin challenge. Compound A presented a good pharmacokinetic property, when given intratracheally, and the blood cells from such pharmacokinetic study showed good receptor occupancy of the compound for approximately 8 hours. Compound A was then tested in an ovalbumin-induced airway inflammation model by intranasal or intravenous route of administration. There was a dose-dependent inhibition of eosinophilia in the bronchiolar lavage fluid, when compound A was given intranasally but not when it was given intravenously. For comparison, antibody to VLA4 and another compound, BIO1211, which reacts only with activated VLA4, were examined in this system. Immunohistochemical analyses of the lung tissue substantiated the findings in the bronchiolar lavage fluid. Specific staining of the major basic protein of eosinophils showed peribronchiolar infiltration of eosinophils. Some of these eosinophils were also positive for nitrotyrosine, suggesting activation of eosinophils in the lung interstitium. There was deposition of major basic protein and nitrotyrosine at the base of the perivascular endothelium, indicative of degranulation of eosinophils in the area. After intranasal treatment with compound A, eosinophils in the lungs and their activation products were substantially decreased, documenting its effectiveness in inhibiting lung inflammation. PMID- 12569080 TI - Role of the quorum-sensing system in experimental pneumonia due to Pseudomonas aeruginosa in rats. AB - The virulence of Pseudomonas aeruginosa is partly controlled by the las quorum sensing system. A rat model of acute pneumonia was used to investigate the pathophysiological impact of this system by comparing the virulence of the wild type virulent laboratory strain PAO1 with that of its lasR-deleted mutant PAOR. In comparison with PAO1, PAOR was avirulent after an instillation of 106 cfu (mortality rates, 72 versus 0%, respectively; p < 0.0001). A ten-fold higher inoculum slightly increased the mortality rate induced by PAOR (25%), which remained lower than that induced by PAO1 (75%, p = 0.0001). In addition, with both inocula lung and bronchoalveolar lavage bacterial counts were significantly lower in rats infected with PAOR than with PAO1 (p or = mean + 2SD in control) showed a higher basal mean blood pressure (BP) before the clamp (89 +/- 3 vs. 82 +/- 2, P < 0.03) but not a higher glucose infusion rate (GIR) compared with low responders (30 microg/dl), (C + AA; n = 6). The control group (AA) received intravenous saline (n = 6). Femoral arteriovenous blood samples and muscle biopsies were obtained during a primed (2.0 micromol/kg) constant infusion (0.05 micromol. kg(-1). min(-1)) of l-[ring (2)H(5)]phenylalanine before and after ingestion of 15 g of EAA. Hypercortisolemia [36.5 +/- 2.1 (C + AA) vs. 9.0 +/- 1.0 microg/dl (AA)] increased postabsorptive arterial, venous, and muscle intracellular phenylalanine concentrations. Hypercortisolemia also increased postabsorptive and post-EAA insulin concentrations. Net protein balance was blunted (40% lower) following EAA ingestion but remained positive for a greater period of time (60 vs. 180 min) in the C + AA group. Thus, although differences in protein metabolism were evident, EAA ingestion improved muscle protein anabolism during acute hypercortisolemia and may help minimize muscle loss following debilitating injury. PMID- 12569086 TI - Effect of thiazolidinediones and metformin on LDL oxidation and aortic endothelium relaxation in diabetic GK rats. AB - In this study, using GK diabetic rats, we compared the effects of three insulin sensitizers on lipid oxidation and the aortic relaxation response. Eight-week-old rats were treated for 4 wk with either troglitazone or pioglitazone, both of which are thiazolidinediones, or with metformin. Despite the fact that only troglitazone has a similarity in structure to alpha-tocopherol, a potent antioxidant, the level of thiobarbituric acid-reactive substance was lower, and the lag time of the conjugated dienes was longer, in the blood samples from the rats in both troglitazone- and pioglitazone-treated groups. In contrast, another insulin sensitizer, metformin, failed to inhibit the oxidation of blood samples. The aortic vasorelaxation response was increased in both troglitazone- and metformin-treated groups compared with the untreated group. These findings suggest that thiazolidinediones have a beneficial effect on lipid oxidation irrespective of the drug's structural similarity to alpha-tocopherol. It is also suggested that the thiazolidinediones and metformin improve vascular function in diabetes. These effects may play a role in the prevention of atherosclerosis in diabetic patients. PMID- 12569087 TI - Lysine synthesized by the gastrointestinal microflora of pigs is absorbed, mostly in the small intestine. AB - This study used a digesta transfer protocol to determine the site of absorption of lysine synthesized by the gastrointestinal microflora of pigs. Eight pigs were used, four with reentrant cannulas in the terminal ileum, two with simple T cannulas in the terminal ileum, and two intact. All pigs were given, for 5 days, the same low-protein diet that included fermentable carbohydrates. The diet of two pigs with reentrant cannulas (donor) and of the two intact (control) pigs was supplemented with (15)NH(4)Cl. The two other pigs with reentrant cannulas (acceptor pigs) and those with simple cannulas (used to supply unlabeled digesta) were given the same diet but unlabeled NH(4)Cl. Ileal digesta were collected continuously from all of the reentrant cannulas and kept on ice. All digesta from each donor pig were reheated and returned to the distal cannula of its companion acceptor, whose ileal digesta were discarded. Unlabeled ileal digesta from the pigs with simple cannulas were instilled into the distal cannulas of the donor pigs. At the end of the experiment, the average (15)N enrichment in the plasma free lysine of control pigs was 0.0407 atom % excess (APE); that of donor pigs was 0.0322 APE (79% of controls), whereas that of acceptor pigs was only 0.0096 APE (24% of controls). Due to nitrogen recycling, acceptor pigs had labeled lysine in the digesta of the stomach and small intestine, and donor pigs had labeled lysine in the digesta of the large intestine. If account is taken of the higher (15)N enrichment of microbial lysine in the large compared with the small intestine, it can be estimated that >90% of the absorption of microbial lysine took place in the small intestine. PMID- 12569088 TI - Effect of intraportal glucagon-like peptide-1 on glucose metabolism in conscious dogs. AB - Arteriovenous difference and tracer ([3-(3)H]glucose) techniques were used in 42 h-fasted conscious dogs to identify any insulin-like effects of intraportally administered glucagon-like peptide 1-(7-36)amide (GLP-1). Each study consisted of an equilibration, a basal, and three 90-min test periods (P1, P2, and P3) during which somatostatin, intraportal insulin (3-fold basal) and glucagon (basal), and peripheral glucose were infused. Saline was infused intraportally in P1. During P2 and P3, GLP-1 was infused intraportally at 0.9 and 5.1 pmol. kg(-1). min(-1) in eight dogs, at 10 and 20 pmol. kg(-1). min(-1) in seven dogs, and at 0 pmol. kg(-1). min(-1) in eight dogs (control group). Net hepatic glucose uptake was significantly enhanced during GLP-1 infusion at 20 pmol. kg(-1). min(-1) [21.8 vs. 13.4 micromol. kg(-1). min(-1) (control), P < 0.05]. Glucose utilization was significantly increased during infusion at 10 and 20 pmol. kg(-1). min(-1) [87.3 +/- 8.3 and 105.3 +/- 12.8, respectively, vs. 62.2 +/- 5.3 and 74.7 +/- 7.4 micromol. kg(-1). min(-1) (control), P < 0.05]. The glucose infusion rate required to maintain hyperglycemia was increased (P < 0.05) during infusion of GLP-1 at 5.1, 10, and 20 pmol. kg(-1). min(-1) (22, 36, and 32%, respectively, greater than control). Nonhepatic glucose uptake increased significantly during delivery of GLP-1 at 5.1 and 10 pmol. kg(-1). min(-1) (25 and 46% greater than control) and tended (P = 0.1) to increase during GLP-1 infusion at 20 pmol. kg( 1). min(-1) (24% greater than control). Intraportal infusion of GLP-1 at high physiological and pharmacological rates increased glucose disposal primarily in nonhepatic tissues. PMID- 12569089 TI - Retraction. Dynein, dynactin, and kinesin II's interaction with microtubules is regulated during bidirectional organelle transport. PMID- 12569090 TI - Phosphorylation of serine 1106 in the catalytic domain of topoisomerase II alpha regulates enzymatic activity and drug sensitivity. AB - Topoisomerases alter DNA topology and are vital for the maintenance of genomic integrity. Topoisomerases I and II are also targets for widely used antitumor agents. We demonstrated previously that in the human leukemia cell line, HL-60, resistance to topoisomerase (topo) II-targeting drugs such as etoposide is associated with site-specific hypophosphorylation of topo II alpha. This effect can be mimicked in sensitive cells treated with the intracellular Ca(2+) chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM). Here we identify Ser-1106 as a major phosphorylation site in the catalytic domain of topo II alpha. This site lies within the consensus sequence for the acidotrophic kinases, casein kinase I and casein kinase II. Mutation of serine 1106 to alanine (S1106A) abrogates phosphorylation of phosphopeptides that were found to be hypophosphorylated in resistant HL-60 cells or sensitive cells treated with BAPTA-AM. Purified topo II alpha containing a S1106A substitution is 4-fold less active than wild type topo II alpha in decatenating kinetoplast DNA and also exhibits a 2-4-fold decrease in the level of etoposide-stabilized DNA cleavable complex formation. Saccharomyces cerevisiae (JN394t2-4) cells expressing S1106A mutant topo II alpha protein are more resistant to the cytotoxic effects of etoposide or amsacrine. These results demonstrate that Ca(2+)-regulated phosphorylation of Ser-1106 in the catalytic domain of topo II alpha modulates the enzymatic activity of this protein and sensitivity to topo II targeting drugs. PMID- 12569091 TI - Ectopic expression of Axin blocks neuronal differentiation of embryonic carcinoma P19 cells. AB - Axin regulates Wnt signaling through down-regulation of beta-catenin. To test the role of Wnt signaling in neuronal differentiation, embryonal carcinoma P19 cells (P19 EC), which can be stimulated to differentiate into a neuron-like phenotype in response to retinoic acid (RA), were used. Reverse transcription-PCR and Western blot analysis showed that Axin is expressed in undifferentiated cells, whereas the level is clearly reduced during RA-induced neuronal differentiation. Interestingly, Axin levels were not reduced during endodermal differentiation of P19 EC cells and F9 EC cells by RA, suggesting that the reduction of the Axin level is a specific property of neuronal differentiation. Western analysis showed that the cytoplasmic level of beta-catenin increased during neuronal differentiation of P19 EC cells. Indirect immunofluorescence with beta-catenin antibody showed that the localization of beta-catenin was changed from membrane in undifferentiated cells to nuclei in neuronal P19 EC cells. Induced expression of Axin during endodermal and early neuronal differentiation, using the Tet-On system, did not block normal differentiation. However, maintenance of the Axin level blocked neuronal differentiation and inhibited expression of a neuron specific marker protein, beta III-tubulin. Also, ectopic induction of a beta catenin signaling inhibitor, ICAT, inhibited expression of beta III-tubulin. In contrast, addition of Wnt-3A-conditioned medium during the neuronal differentiation period enhanced the expression of beta III-tubulin. Overall, our data show that Wnt-3a/canonical beta-catenin signaling through the down regulation of Axin may play an important role in neuronal differentiation. PMID- 12569092 TI - Transcription factor IID recruitment and Sp1 activation. Dual function of TAF1 in cyclin D1 transcription. AB - Cyclin D1 is an oncogene that regulates progression through the G(1) phase of the cell cycle. A temperature-sensitive missense mutation in the transcription factor TAF1/TAF(II)250 induces the mutant ts13 cells to arrest in late G(1) by decreasing transcription of cell cycle regulators, including cyclin D1. Here we provide evidence that TAF1 serves two independent functions, one at the core promoter and one at the upstream activating Sp1 sites of the cyclin D1 gene. Using in vivo genomic footprinting, we have identified protein-DNA interactions within the cyclin D1 core promoter that are disrupted upon inactivation of TAF1 in ts13 cells. This 33-bp segment, which we termed the TAF1-dependent element 1 (TDE1), contains an initiation site that displays homology to the consensus motif and is sufficient to confer a requirement for TAF1 function. Electrophoretic mobility shift assays reveal that binding of ts13-TAF1-containing TFIID complexes to the cyclin D1 TDE1 occurs at 25 degrees C but not at 37 degrees C in vitro and involves the initiator element. Temperature-dependent DNA binding activity is also observed for TAF1-TAF2 heterodimers assembled with the ts13 mutant but not the wild-type TAF1 protein. These data suggest that a function of TAF is required for the interaction of TFIID with the cyclin D1 initiator. Our finding that recruitment of TFIID, by insertion of a TBP binding site upstream of the TDE1, restores basal but not activated transcription supports the model that TAF1 carries out two independent functions at the cyclin D1 promoter. PMID- 12569093 TI - Functional interaction between the c-Abl and Arg protein-tyrosine kinases in the oxidative stress response. AB - The Abl family of mammalian nonreceptor tyrosine kinases consists of c-Abl and Arg. Recent work has shown that c-Abl and Arg are activated in the cellular response to oxidative stress. The present studies demonstrate that reactive oxygen species (ROS) induce the formation of c-Abl and Arg heterodimers. The results show that the c-Abl SH3 domain binds directly to a proline-rich site (amino acids 567-576) in the Arg C-terminal region. Formation of c-Abl.Arg heterodimers also involves direct binding of the Arg Src homology 3 domain to the C-terminal region of c-Abl. The results further demonstrate that the interaction between c-Abl and Arg involves c-Abl-mediated phosphorylation of Arg. The functional significance of the c-Abl-Arg interaction is supported by the demonstration that both c-Abl and Arg are required for ROS-induced apoptosis. These findings indicate that ROS induce c-Abl.Arg heterodimers and that both c Abl and Arg are necessary as effectors in the apoptotic response to oxidative stress. PMID- 12569094 TI - The role of the minor groove substituents in indirect readout of DNA sequence by 434 repressor. AB - The sequence of non-contacted bases at the center of the 434 repressor binding site affects the strength of the repressor-DNA complex by influencing the structure and flexibility of DNA (Koudelka, G. B., and Carlson, P. (1992) Nature 355, 89-91). We synthesized 434 repressor binding sites that differ in their central sequence base composition to test the importance of minor groove substituents and/or the number of base pair hydrogen bonds between these base pairs on DNA structure and strength of the repressor-DNA complex. We show here that the number of base pair H-bonds between the central bases apparently has no role in determining the relative affinity of a DNA site for repressor. Instead we find that the affinity of DNA for repressor depends on the absence or presence the N2-NH(2) group on the purine bases at the binding site center. The N2-NH(2) group on bases at the center of the 434 binding site appears to destabilize 434 repressor-DNA complexes by decreasing the intimacy of the specific repressor-DNA contacts, while increasing the reliance on protein contacts to the DNA phosphate backbone. Thus, the presence of an N2-NH(2) group on the purines at the center of a binding site globally alters the precise conformation of the protein-DNA interface. PMID- 12569095 TI - An NMR-based model of the ubiquitin-bound human ubiquitin conjugation complex Mms2.Ubc13. The structural basis for lysine 63 chain catalysis. AB - A heterodimer composed of the catalytically active ubiquitin-conjugating enzyme hUbc13 and its catalytically inactive paralogue, hMms2, forms the catalytic core for the synthesis of an alternative type of multiubiquitin chain where ubiquitin molecules are tandemly linked to one another through a Lys-63 isopeptide bond. This type of linkage, as opposed to the more typical Lys-48-linked chains, serves as a non-proteolytic marker of protein targets involved in error-free post replicative DNA repair and NF-kappa B signal transduction. Using a two dimensional (1)H-(15)N NMR approach, we have mapped: 1) the interaction between the subunits of the human Ubc13.Mms2 heterodimer and 2) the interactions between each of the subunits or heterodimer with a non-covalently bound acceptor ubiquitin or a thiolester-linked donor ubiquitin. Using these NMR-derived constraints and an unbiased docking approach, we have assembled the four components of this catalytic complex into a three-dimensional model that agrees well with its catalytic function. PMID- 12569096 TI - Mutational analysis of the non-homologous region of subunit A of the yeast V ATPase. AB - Subunit A is the catalytic nucleotide binding subunit of the vacuolar proton translocating ATPase (or V-ATPase) and is homologous to subunit beta of the F(1)F(0) ATP synthase (or F-ATPase). Amino acid sequence alignment of these subunits reveals a 90-amino acid insert in subunit A (termed the non-homologous region) that is absent from subunit beta. To investigate the functional role of this region, site-directed mutagenesis has been performed on the VMA1 gene that encodes subunit A in yeast. Substitutions were performed on 13 amino acid residues within this region that are conserved in all available A subunit sequences. Most of the 18 mutations introduced showed normal assembly of the V ATPase. Of these, one (R219K) greatly reduced both proton transport and ATPase activity. By contrast, the P217V mutant showed significantly reduced ATPase activity but higher than normal levels of proton transport, suggesting an increase in coupling efficiency. Two other mutations in the same region (P223V and P233V) showed decreased coupling efficiency, suggesting that changes in the non-homologous region can alter coupling of proton transport and ATP hydrolysis. It was previously shown that the V-ATPase must possess at least 5-10% activity relative to wild type to undergo in vivo dissociation in response to glucose withdrawal. However, four of the mutations studied (G150A, D157E, P177V, and P223V) were partially or completely blocked in dissociation despite having greater than 30% of wild type levels of activity. These results suggest that changes in the non-homologous region can also alter in vivo dissociation of the V ATPase independent of effects on activity. PMID- 12569097 TI - Transition from natively unfolded to folded state induced by desiccation in an anhydrobiotic nematode protein. AB - Late embryogenesis abundant (LEA) proteins are associated with desiccation tolerance in resurrection plants and in plant seeds, and the recent discovery of a dehydration-induced Group 3 LEA-like gene in the nematode Aphelenchus avenae suggests a similar association in anhydrobiotic animals. Despite their importance, little is known about the structure of Group 3 LEA proteins, although computer modeling and secondary structure algorithms predict a largely alpha helical monomer that forms coiled coil oligomers. We have therefore investigated the structure of the nematode protein, AavLEA1, in the first such analysis of a well characterized Group 3 LEA-like protein. Immunoblotting and subunit cross linking experiments demonstrate limited oligomerization of AavLEA1, but analytical ultracentrifugation and gel filtration show that the vast majority of the protein is monomeric. Moreover, CD, fluorescence emission, and Fourier transform-infrared spectroscopy indicate an unstructured conformation for the nematode protein. Therefore, in solution, no evidence was found to support structure predictions; instead, AavLEA1 seems to be natively unfolded with a high degree of hydration and low compactness. Such proteins can, however, be induced to fold into more rigid structures by partner molecules or by altered physiological conditions. Because AavLEA1 is associated with desiccation stress, its Fourier transform-infrared spectrum in the dehydrated state was examined. A dramatic but reversible increase in alpha-helix and, possibly, coiled coil formation was observed on drying, indicating that computer predictions of secondary structure may be correct for the solid state. This unusual finding offers the possibility that structural shifts in Group 3 LEA proteins occur on dehydration, perhaps consistent with their role in anhydrobiosis. PMID- 12569098 TI - Identification of an archaeal alpha-L-fucosidase encoded by an interrupted gene. Production of a functional enzyme by mutations mimicking programmed -1 frameshifting. AB - The analysis of the complete genome of the thermoacidophilic Archaeon Sulfolobus solfataricus revealed two open reading frames (ORF), named SSO11867 and SSO3060, interrupted by a -1 frameshift and encoding for the N- and the C-terminal fragments, respectively, of an alpha-l-fucosidase. We report here that these ORFs are actively transcribed in vivo, and we confirm the presence of the -1 frameshift between them at the cDNA level, explaining why we could not find alpha fucosidase activity in S. solfataricus extracts. Detailed analysis of the region of overlap between the two ORFs revealed the presence of the consensus sequence for a programmed -1 frameshifting. Two specific mutations, mimicking this regulative frameshifting event, allow the expression, in Escherichia coli, of a fully active thermophilic and thermostable alpha-l-fucosidase (EC ) with micromolar substrate specificity and showing transfucosylating activity. The analysis of the fucosylated products of this enzyme allows, for the first time, assigning a retaining reaction mechanism to family 29 of glycosyl hydrolases. The presence of an alpha-fucosidase putatively regulated by programmed -1 frameshifting is intriguing both with respect to the regulation of gene expression and, in post-genomic era, for the definition of gene function in Archaea. PMID- 12569100 TI - Crystal structure of two ternary complexes of phosphorylating glyceraldehyde-3 phosphate dehydrogenase from Bacillus stearothermophilus with NAD and D glyceraldehyde 3-phosphate. AB - The crystal structure of the phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus was solved in complex with its cofactor, NAD, and its physiological substrate, D-glyceraldehyde 3-phosphate (D-G3P). To isolate a stable ternary complex, the nucleophilic residue of the active site, Cys(149), was substituted with alanine or serine. The C149A and C149S GAPDH ternary complexes were obtained by soaking the crystals of the corresponding binary complexes (enzyme.NAD) in a solution containing G3P. The structures of the two binary and the two ternary complexes are presented. The D G3P adopts the same conformation in the two ternary complexes. It is bound in a non-covalent way, in the free aldehyde form, its C-3 phosphate group being positioned in the P(s) site and not in the P(i) site. Its C-1 carbonyl oxygen points toward the essential His(176), which supports the role proposed for this residue along the two steps of the catalytic pathway. Arguments are provided that the structures reported here are representative of a productive enzyme.NAD.D-G3P complex in the ground state (Michaelis complex). PMID- 12569099 TI - N-oleoyldopamine, a novel endogenous capsaicin-like lipid that produces hyperalgesia. AB - N-Arachidonoyldopamine (NADA) was recently identified as an endogenous ligand for the vanilloid type 1 receptor (VR1). Further analysis of the bovine striatal extract from which NADA was isolated indicated the existence of substances corresponding in molecular mass to N-oleoyldopamine (OLDA), N-palmitoyldopamine (PALDA), and N-stearoyldopamine (STEARDA). Quadrupole time-of-flight mass spectrometric analysis of bovine striatal extracts revealed the existence of OLDA, PALDA, and STEARDA as endogenous compounds in the mammalian brain. PALDA and STEARDA failed to affect calcium influx in VR1-transfected human embryonic kidney (HEK) 293 cells or paw withdrawal latencies from a radiant heat source, and there was no evidence of spontaneous pain behavior. By contrast, OLDA induced calcium influx (EC(50) = 36 nm), reduced the latency of paw withdrawal from a radiant heat source in a dose-dependent manner (EC(50) = 0.72 microg), and produced nocifensive behavior. These effects were blocked by co-administration of the VR1 antagonist iodo-resiniferatoxin (10 nm for HEK cells and 1 microg/50 micro;l for pain behavior). These findings demonstrate the existence of an endogenous compound in the brain that is similar to capsaicin and NADA in its chemical structure and activity on VR1. Unlike NADA, OLDA was only a weak ligand for rat CB1 receptors; but like NADA, it was recognized by the anandamide membrane transporter while being a poor substrate for fatty-acid amide hydrolase. Analysis of the activity of six additional synthetic and potentially endogenous N acyldopamine indicated the requirement of a long unsaturated fatty acid chain for an optimal functional interaction with VR1 receptors. PMID- 12569101 TI - Mutational analysis of TraR. Correlating function with molecular structure of a quorum-sensing transcriptional activator. AB - TraR, the quorum-sensing activator of the Agrobacterium tumefaciens Ti plasmid conjugation system, induces gene expression in response to its quormone, N-(3 oxooctanoyl)-L-homoserine lactone. Ligand binding results in dimerization of TraR and is required for its activity. Analysis of N- and C-terminal deletion mutants of TraR localized the quormone-binding domain to a region between residues 39 and 140 and the primary dimerization domain to a region between residues 119 and 156. The dominant-negative properties of these mutants predicted a second dimerization domain at the C terminus of the protein. Analysis of fusions of N-terminal fragments of TraR to lambda cI' confirmed the dimerization activity of these two domains. Fifteen single amino acid substitution mutants of TraR defective in dimerization were isolated. According to the analysis of these mutants, Asp-70 and Gly-113 are essential for quormone binding, whereas Ala-38 and Ala-105 are important, but not essential. Additional residues located within the N-terminal half of TraR, including three located in alpha-helix 9, contribute to dimerization, but are not required for ligand binding. These results and the recently reported crystal structure of TraR are consistent with and complement each other and together define some of the structural and functional relationships of this quorum-sensing activator. PMID- 12569102 TI - Sustained induction of ERK, protein kinase B, and p70 S6 kinase regulates cell spreading and formation of F-actin microspikes upon ligation of integrins by galectin-8, a mammalian lectin. AB - Galectin-8, a member of the galectin family of mammalian lectins, is a secreted protein that promotes cell adhesion and migration upon binding to a subset of integrins through sugar-protein interactions. Ligation of integrins by galectin-8 triggers a distinct pattern of cytoskeletal organization, including formation of F-actin-containing microspikes. This is associated with activation of integrin mediated signaling cascades (ERK and phosphatidylinositol 3 kinase (PI3K)) that are much more robust and are of longer duration than those induced upon cell adhesion to fibronectin. Indeed, formation of microspikes is enhanced 40% in cells that overexpress protein kinase B, the downstream effector of PI3K. Inhibition of PI3K activity induced by wortmannin partially inhibits cell adhesion and spreading while largely inhibiting microspike formation in cells adherent to galectin-8. Furthermore, the inhibitory effects of wortmannin are markedly accentuated in cells overexpressing PKB or p70S6K (CHO(PKB) and CHO(p70S6K) cells), whose adhesion and spreading on galectin-8 (but not on fibronectin) is inhibited approximately 25-35% in the presence of wortmannin. The above results suggest that galectin-8 is an extracellular matrix protein that triggers a unique repertoire of integrin-mediated signals, which leads to a distinctive cytoskeletal organization and microspike formation. They further suggest that downstream effectors of PI3K, including PKB and p70 S6 kinase, in part mediate cell adhesion, spreading, and microspike formation induced by galectin-8. PMID- 12569103 TI - Characterization of a functional bacterial homologue of sodium-dependent neurotransmitter transporters. AB - The tnaT gene of Symbiobacterium thermophilum encodes a protein homologous to sodium-dependent neurotransmitter transporters. Expression of the tnaT gene product in Escherichia coli conferred the ability to accumulate tryptophan from the medium and the ability to grow on tryptophan as a sole source of carbon. Transport was Na(+)-dependent and highly selective. The K(m) for tryptophan was approximately 145 nm, and tryptophan transport was unchanged in the presence of 100 microM concentrations of other amino acids. Tryptamine and serotonin were weak inhibitors with K(I) values of 200 and 440 microM, respectively. By using a T7 promoter-based system, TnaT with an N-terminal His(6) tag was expressed at high levels in the membrane and was purified to near-homogeneity in high yield. PMID- 12569104 TI - Isolation, structural characterization, and properties of mattacin (polymyxin M), a cyclic peptide antibiotic produced by Paenibacillus kobensis M. AB - Mattacin is a nonribosomally synthesized, decapeptide antibiotic produced by Paenibacillus kobensis M. The producing strain was isolated from a soil/manure sample and identified using 16 S rRNA sequence homology along with chemical and morphological characterization. An efficient production and isolation procedure was developed to afford pure mattacin. Structure elucidation using a combination of chemical degradation, multidimensional NMR studies (COSY, HMBC, HMQC, ROESY), and mass spectrometric (MALDI MS/MS) analyses showed that mattacin is identical to polymyxin M, an uncommon antibiotic reported previously in certain Bacillus species by Russian investigators. Mattacin (polymyxin M) is cyclic and possesses an amide linkage between the C-terminal threonine and the side chain amino group of the diaminobutyric acid residue at position 4. It contains an (S)-6 methyloctanoic acid moiety attached as an amide at the N-terminal amino group, one D-leucine, six L-alpha,gamma-diaminobutyric acid, and three L-threonine residues. Transfer NOE experiments on the conformational preferences of mattacin when bound to lipid A and microcalorimetry studies on binding to lipopolysaccharide showed that its behavior was very similar to that observed in previous studies of polymyxin B (a commercial antibiotic), suggesting an identical mechanism of action. It was capable of inhibiting the growth of a wide variety of Gram-positive and Gram-negative bacteria, including several human and plant pathogens with activity comparable with purified polymyxin B. The biosynthesis of mattacin was also examined briefly using transpositional mutagenesis by which 10 production mutants were obtained, revealing a set of genes involved in production. PMID- 12569105 TI - Plicatamide, an antimicrobial octapeptide from Styela plicata hemocytes. AB - Plicatamide (Phe-Phe-His-Leu-His-Phe-His-dc Delta DOPA), where dc Delta DOPA represents decarboxy-(E)-alpha,beta-dehydro-3,4-dihydroxyphenylalanine, is a potently antimicrobial octapeptide from the blood cells of the solitary tunicate, Styela plicata. Wild type and methicillin-resistant Staphylococcus aureus (MRSA) responded to plicatamide exposure with a massive potassium efflux that began within seconds. Soon thereafter, treated bacteria largely ceased consuming oxygen, and most became nonviable. Native plicatamide also formed cation selective channels in model lipid bilayers composed of bacterial lipids. Methicillin-resistant S. aureus treated with plicatamide for 5 min contained prominent mesosomes as well as multiple, small dome-shaped surface protrusions that suggested the involvement of osmotic forces in its antimicrobial effects. To ascertain the contribution of the C-terminal dc Delta DOPA residue to antimicrobial activity, we synthesized several analogues of plicatamide that lacked it. One of these peptides, PL-101 (Phe-Phe-His-Leu-His-Phe-His-Tyr-amide), closely resembled native plicatamide in its antimicrobial activity and its ability to induce potassium efflux. Plicatamide was potently hemolytic for human red blood cells but did not lyse ovine erythrocytes. The small size, rapid action, and potent anti-staphylococcal activity of plicatamide and PL-101 make them intriguing subjects for future antimicrobial peptide design. PMID- 12569106 TI - Purification and characterization of monolysocardiolipin acyltransferase from pig liver mitochondria. AB - In mammalian tissues cardiolipin is rapidly remodeled by monolysocardiolipin acyltransferase subsequent to its de novo biosynthesis (Ma, B. J., Taylor, W. A, Dolinsky, V. W., and Hatch, G. M. (1999) J. Lipid Res. 40, 1837-1845). We report here the purification and characterization of a monolysocardiolipin acyltransferase activity from pig liver mitochondria. Monolysocardiolipin acyltransferase activity was purified over 1000-fold by butanol extraction, hydroxyapatite chromatography, and preparative SDS-PAGE. The purified 74-kDa protein catalyzed acylation of monolysocardiolipin to cardiolipin with [(14)C]linoleoyl coenzyme A. Photoaffinity labeling of the protein with 12-[(4 [(125)I]azidosalicyl)amino]dodecanoyl coenzyme A indicated coenzyme A was bound at its active site and photoaffinity cross-linking of 12-[(4 azidosalicyl)amino]dodecanoyl coenzyme A to the enzyme inhibited enzyme activity. Enzyme activity was optimum at pH 7.0, and the enzyme did not utilize other lysophospholipids as substrate. The purified enzyme was heat-labile and exhibited an isoelectric point of pH 5.4. To determine the enzymes kinetic mechanism the effect of varying concentrations of linoleoyl coenzyme A and monolysocardiolipin on initial velocity were determined. Double-reciprocal plots revealed parallel lines consistent with a ping pong kinetic mechanism. When the enzyme was incubated in the absence of monolysocardiolipin, coenzyme A was produced from linoleoyl coenzyme A at a rate consistent with the formation of an enzyme linoleate intermediate. The true K(m) value for linoleoyl coenzyme A and true K(m) value for monolysocardiolipin were 100 and 44 microM, respectively. The calculated V(max) was 6802 pmol/min per mg of protein. A polyclonal antibody, raised in rabbits to the purified protein, cross-reacted with the protein in crude pig liver mitochondrial fractions. In liver mitochondria prepared from thyroxine-treated rats, the level of the protein was elevated compared with euthyroid controls indicating that expression of monolysocardiolipin acyltransferase is regulated by thyroid hormone. The study represents the first purification and characterization of a monolysocardiolipin acyltransferase activity from any organism. PMID- 12569107 TI - Characterization of human thioredoxin-like 2. A novel microtubule-binding thioredoxin expressed predominantly in the cilia of lung airway epithelium and spermatid manchette and axoneme. AB - We describe here the cloning and characterization of a novel member of the thioredoxin family, thioredoxin-like protein 2 (Txl-2). The Txl-2 open reading frame codes for a protein of 330 amino acids consisting of two distinct domains: an N-terminal domain typical of thioredoxins and a C-terminal domain belonging to the nucleoside-diphosphate kinase family, separated by a small interface domain. The Txl-2 gene spans approximately 28 kb, is organized into 11 exons, and maps at locus 3q22.3-q23. A splicing variant lacking exon 5 (Delta 5Txl-2) has also been isolated. By quantitative real time PCR we demonstrate that Txl-2 mRNA is ubiquitously expressed, with testis and lung having the highest levels of expression. Unexpectedly, light and electron microscopy analyses show that the protein is associated with microtubular structures such as lung airway epithelium cilia and the manchette and axoneme of spermatids. Using in vitro translated proteins, we demonstrate that full-length Txl-2 weakly associates with microtubules. In contrast, Delta 5Txl-2 specifically binds with very high affinity brain microtubule preparations containing microtubule-binding proteins. Importantly, Delta 5Txl-2 also binds to pure microtubules, proving that it possesses intrinsic microtubule binding capability. Taken together, Delta 5Txl-2 is the first thioredoxin reported to bind microtubules and might therefore be a novel regulator of microtubule physiology. PMID- 12569108 TI - Inactivation of Cdc13p triggers MEC1-dependent apoptotic signals in yeast. AB - Inactivation of the budding yeast telomere binding protein Cdc13 results in abnormal telomeres (exposed long G-strands) and activation of the DNA damage checkpoint. In the current study, we show that inactivation of Cdc13p induces apoptotic signals in yeast, as evidenced by caspase activation, increased reactive oxygen species production, and flipping of phosphatidylserine in the cytoplasmic membrane. These apoptotic signals were suppressed in a mitochondrial (rho(o)) mutant. Moreover, mitochondrial proteins (e.g. MTCO3) were identified as multicopy suppressors of cdc13-1, suggesting the involvement of mitochondrial functions in telomere-initiated apoptotic signaling. These telomere-initiated apoptotic signals were also shown to depend on MEC1, but not TEL1, and were antagonized by MRE11. Our results are consistent with a model in which single stranded G-tails in the cdc13-1 mutant trigger MEC1-dependent apoptotic signaling in yeast. PMID- 12569109 TI - Folate: a key to optimizing health and reducing disease risk in the elderly. AB - Inadequate folate status is associated with an increased risk for chronic diseases that may have a negative impact on the health of the aging population. Folate, a water-soluble vitamin, includes naturally occurring food folate and synthetic folic acid in supplements and fortified foods. Inadequate folate status may result in hyperhomocysteinemia, a significant risk factor for atherosclerotic vascular disease, changes in DNA that may result in pro-carcinogenic effects and increased risk for cognitive dysfunction. Folate status may be negatively influenced by inadequate intake, genetic polymorphisms and interactions with various drugs. In the US, folic acid is now added to enriched grain products and continues to be included in the majority of ready-to-eat breakfast cereals. Recent data indicate that the folate status in the US population has improved significantly, presumably due to the effects of fortification. Folic acid (not food folate) intake in excess of the Tolerable Upper Intake Level may mask the diagnosis of a vitamin B(12) deficiency, which is more prevalent in the elderly than younger individuals. When folic acid supplements are recommended, a multivitamin that includes vitamin B(12) should also be advised. To safely and effectively increase folate intake in the elderly, naturally occurring folate rich food sources should be promoted. Folate-rich foods include orange juice, dark green leafy vegetables, asparagus, strawberries and legumes. These foods are also excellent sources of other health-promoting nutrients associated with chronic disease risk reduction. PMID- 12569110 TI - The Zone Diet phenomenon: a closer look at the science behind the claims. AB - The purported health benefits of low-carbohydrate diets have been advocated intermittently over the last century and have enjoyed increasing popularity over the last decade. Although most revolve around the emphatic theme that carbohydrates are to blame for many chronic diseases, their specific ideologies are more variable and in some cases quite sophisticated. The Zone Diet phenomenon represents a new generation of modern low carbohydrate food fad with sales placing it among the most popular diet books in recent history. The Zone is a 40% carbohydrate, 30% protein and 30% fat eating plan that advocates only sparing use of grains and starches. The precise 0.75 protein to carbohydrate ratio required with each meal is promoted to reduce the insulin to glucagon ratio, which purportedly affects eicosanoid metabolism and ultimately produces a cascade of biological events leading to a reduction in chronic disease risk, enhanced immunity, maximal physical and mental performance, increased longevity and permanent weight loss. There is presently little scientific support for the connections made between diet, endocrinology and eicosanoid metabolism. In fact, a review of the literature suggests that there are scientific contradictions in the Zone Diet hypothesis that cast unquestionable doubt on its potential efficacy. The purpose of this review is to evaluate the scientific merit of the Zone Diet and its health claims in an effort to help delineate what is and what is not sound nutrition science. PMID- 12569111 TI - Vitamin C as an antioxidant: evaluation of its role in disease prevention. AB - Vitamin C in humans must be ingested for survival. Vitamin C is an electron donor, and this property accounts for all its known functions. As an electron donor, vitamin C is a potent water-soluble antioxidant in humans. Antioxidant effects of vitamin C have been demonstrated in many experiments in vitro. Human diseases such as atherosclerosis and cancer might occur in part from oxidant damage to tissues. Oxidation of lipids, proteins and DNA results in specific oxidation products that can be measured in the laboratory. While these biomarkers of oxidation have been measured in humans, such assays have not yet been validated or standardized, and the relationship of oxidant markers to human disease conditions is not clear. Epidemiological studies show that diets high in fruits and vegetables are associated with lower risk of cardiovascular disease, stroke and cancer, and with increased longevity. Whether these protective effects are directly attributable to vitamin C is not known. Intervention studies with vitamin C have shown no change in markers of oxidation or clinical benefit. Dose concentration studies of vitamin C in healthy people showed a sigmoidal relationship between oral dose and plasma and tissue vitamin C concentrations. Hence, optimal dosing is critical to intervention studies using vitamin C. Ideally, future studies of antioxidant actions of vitamin C should target selected patient groups. These groups should be known to have increased oxidative damage as assessed by a reliable biomarker or should have high morbidity and mortality due to diseases thought to be caused or exacerbated by oxidant damage. PMID- 12569112 TI - Konjac supplement alleviated hypercholesterolemia and hyperglycemia in type 2 diabetic subjects--a randomized double-blind trial. AB - OBJECTIVES: The present study was designed to evaluate effects of konjac glucomannan (KGM) supplement (3.6 g/day) for 28 days on blood lipid and glucose levels in hyperlipidemic type 2 diabetic patients and the possible mechanism for the reductions in blood lipid levels. METHODS: Twenty-two diabetic subjects (age 64.2 + 8.4 years, BMI 25.5 + 3.2 kg/m(2)) with elevated blood cholesterol levels (fasting glucose between 6.7-14.4 mmol/L), but currently not taking lipid lowering medication, were recruited to participate in a two 28-day period, randomized, double-blind, crossover clinical trial. Fasting blood samples drawn on the initial and final days of each period were determined for plasma lipids and glucose levels. Feces collected at the end of each experimental period were analyzed for neutral sterol and bile acid contents. RESULTS: Compared with placebo, KGM effectively reduced plasma cholesterol (11.1%, p = 0.0001, adjusted alpha = 0.006), LDL-cholesterol (20.7%, p = 0.0004, adjusted alpha = 0.006), total/HDL cholesterol ratio (15.6%, p = 0.0005, adjusted alpha = 0.007), ApoB (12.9%, p = 0.0001, adjusted alpha = 0.006) and fasting glucose (23.2%, p = 0.002, adjusted alpha = 0.008). Plasma triglyceride, HDL-cholesterol, LDL/HDL cholesterol, postprandial glucose and body weight were not significant after adjustment by the Bonferroni-Hochberg procedure. Fecal neutral sterol and bile acid concentrations were increased by 18.0% (p = 0.004) and 75.4% (p < 0.001), respectively, with KGM supplement. CONCLUSIONS: The KGM supplement improved blood lipid levels by enhancing fecal excretion of neutral sterol and bile acid and alleviated the elevated glucose levels in diabetic subjects. KGM could be an adjunct for the treatment of hyperlipidemic diabetic subjects. PMID- 12569113 TI - Plasma selenium and plasma and erythrocyte glutathione peroxidase activity increase with estrogen during the menstrual cycle. AB - OBJECTIVE: This study was designed to determine the timing and magnitude of changes in selenium status in relation to the fluctuation of 17-beta-estradiol during the menstrual cycle and the effect of different phases of the menstrual cycle on dietary selenium intake. METHODS: Plasma 17-beta-estradiol and plasma and erythrocyte selenium and glutathione peroxidase (GPx) activity were measured in fasting blood samples collected in the morning at four times over three phases of the menstrual cycle: early follicular (EF: days 1-3 menstruation), periovulatory (PO; E-1: 1 day before estrogen peak and E: during estrogen peak) and mid-luteal (ML: 7-9 days after ovulation) in healthy women (n = 14) aged 21 to 39 years and with regular menstrual cycles (26 to 30 days). The estrogen peak was confirmed by measurement of the luteinizing hormone surge. Dietary records (three-day) coincided with blood collection for each phase. RESULTS: Plasma selenium and plasma and erythrocyte GPx activity were greatest during the periovulatory phase, coinciding with the estrogen peak. No differences were observed for erythrocyte selenium or dietary selenium throughout the cycle. A linear relationship existed between estradiol and plasma selenium (p < 0.0027), plasma GPx activity (p < 0.0001), and erythrocyte GPx activity (p < 0.0001). CONCLUSIONS: These results indicate that blood selenium parameters fluctuate during the menstrual cycle such that the phase of the cycle should be considered when assessing selenium status. PMID- 12569114 TI - Cholesterol vehicle in experimental atherosclerosis 24: avocado oil. AB - OBJECTIVE: To determine atherogenicity of avocado oil relative to saturated (coconut oil), monounsaturated (olive oil) and polyunsaturated (corn oil) fats. METHODS: New Zealand White rabbits were fed a semipurified diet containing 0.2% cholesterol and 14% fat for 90 days. They were then necropsied and severity of atherosclerosis was determined visually. RESULTS: Coconut oil was the most atherogenic fat. Corn oil was only slightly less atherogenic than either olive or avocado oils. Percentage of serum HDL cholesterol was highest in the rabbits fed the two monounsaturated fats. CONCLUSION: Avocado oil is of the same order of atherogenicity as corn oil and olive oil. PMID- 12569115 TI - Randomized controlled trial of the effect of bifidobacteria-fermented milk on ulcerative colitis. AB - BACKGROUND: Alterations of intestinal flora, such as reduction in the concentration of bifidobacteria and increase in that of Bacteroides species, are apparently associated with the severity of ulcerative colitis. OBJECTIVE: We conducted a randomised clinical trial of the use of a bifidobacteria-fermented milk (BFM) supplement as a dietary adjunct in the treatment of ulcerative colitis. METHODS: The subjects were randomly divided into two groups: a group with BFM supplementation (BFM group, 11 subjects) and a control group (control group, 10 subjects). The BFM group was given 100 mL/day of BFM for one year. Colonoscopies, general blood markers and examinations of intestinal flora including the analysis of fecal organic acids were performed at the commencement of the study and after one year. RESULTS: Exacerbation of symptoms was seen in 3 out of 11 subjects in the BFM group and in 9 out of 10 in the control group. Log rank statistic analysis of the cumulative exacerbation rates showed a significant reduction in exacerbations for the BFM group (p = 0.0184). The analysis of microflora and the organic acids in the feces showed a significant reduction in the relative proportion of B. vulgatus in Bacteroidaceae and butyrate concentration, respectively, after supplementation with BFM, in comparison with before. CONCLUSION: Supplementation with the BFM product was successful in maintaining remission and had possible preventive effects on the relapse of ulcerative colitis. PMID- 12569116 TI - Calcium intake of Asian, Hispanic and white youth. AB - OBJECTIVE: To examine calcium intake and food sources among Asian, white and Hispanic youth, in order to develop and target interventions to improve calcium intake. METHODS: Cross-sectional survey with two 24-hour dietary recalls one week apart. Calcium intake was evaluated in 167 male and female adolescents of Asian, Hispanic and white ethnicity, ages 10-18 years, from six states. Main outcome measures were mean daily calcium intake (mg/day). STATISTICAL ANALYSES PERFORMED: t tests, Chi-square and analysis of variance for differences by age, ethnicity and gender, multiple regression of factors influencing calcium intake. RESULTS: Overall median calcium intake was 938 mg/day with 868 mg/day for Asians, 1180 mg/day for whites and 896 mg/day for Hispanics. Daily milk intake was the primary predictor of calcium intake with Asian ethnicity and female gender each showing a negative association to calcium intake in multiple regression models. CONCLUSIONS: Milk intake was the primary factor positively influencing calcium intake, while Asian ethnicity and female gender negatively influenced calcium intake. Thus, interventions to improve calcium intake should focus on improving milk intake of Asians and females. PMID- 12569117 TI - Carbohydrate intake is associated with diet quality and risk factors for cardiovascular disease in U.S. adults: NHANES III. AB - OBJECTIVES: To determine if carbohydrate intake, as a % of energy, was related to diet quality and risk factors for cardiovascular disease (CVD) in adults in a cross-sectional and population-based study in the U.S. METHODS: Data from the third National Health and Nutrition Examination Survey (NHANES III, 1988-1994) were utilized. The nationally representative sample of the U.S. population (3,754 men, 4,074 women, ages 25 to 64 years) was divided into quintiles of carbohydrate intake (% of energy), which was examined in relation to risk factors for CVD: systolic blood pressure, body mass index (BMI), and concentrations of serum triglyceride, serum total and HDL cholesterol and plasma glucose. RESULTS: When covariates (age, ethnicity, smoking, alcohol intake and total energy intake) were adjusted in multivariate analyses, carbohydrate intakes (% of energy) were inversely associated with BMI and serum total cholesterol concentration in men and BMI in women and positively associated with serum triglyceride concentration in women. When total sugar intake (% of energy) was further controlled as a step to understand the quality of carbohydrate, carbohydrate intakes (% of energy) was a stronger predictor of BMI and plasma glucose in men and BMI in women. A high carbohydrate diet (>57.4% of energy in men and >59.1% of energy in women) was associated with a low serum HDL-cholesterol concentration in men and high serum triglyceride in women. CONCLUSION: Moderately high carbohydrate (50% to 55% of energy) diets were associated with low CVD risks with favorable lipid profiles. PMID- 12569118 TI - Postprandial lipemia in hypertension. AB - OBJECTIVE: Many studies have shown that patients with coronary artery disease have an exaggerated rise and a delayed fall of plasma triglyceride (TG) concentration postprandially. We examined whether patients with essential hypertension have the same response to a fatty meal. METHODS: A fatty meal (350g per 2 m(2) body surface with 83.5% fat) was given to 25 patients with essential hypertension (H) and to 25 normotensives (N). The two groups were matched for age, body mass index, lipid profile, basal glucose and insulin concentrations, and an index of homeostasis model of insulin resistance (HOMA-IR). A quantitative insulin sensitivity check index (QUICKI) was calculated. Blood samples were taken at 0, 4, 6, and 8 hours after the fatty meal. Lipid variables were measured in all samples. Blood glucose and insulin levels were measured in the fasting state. RESULTS: Total and high density lipoprotein cholesterol, apolipoprotein A1 and B, lipoprotein (a), HOMA-IR and QUICKI did not differ significantly over time between the groups. The plasma TG concentration (mg/dL) increased significantly after fat loading in H (from 118 +/- 31 to 284 +/- 137 at 4 hours, 327 +/- 93 at 6 hours and 285 +/- 71 at 8 hours) compared to N group (from 105 +/- 29 to 150 +/ 38 at 4 hours, 148 +/- 40 at 6 hours and 115 +/- 34 at 8 hours), p = 0.001, p < 0.001 and p < 0.001, respectively. CONCLUSION: This study suggests that patients with hypertension have an exaggerated response and delayed clearance of plasma TG concentration after fat loading. PMID- 12569119 TI - Caseinphosphopeptides in milk and fermented milk do not affect calcium metabolism acutely in postmenopausal women. AB - BACKGROUND: Caseinphosphopeptides (CPPs) are formed in food processing or during digestion in the gastrointestinal tract. CPPs prevent the formation of insoluble calcium salts; thus, the hypothesis is that CPPs increase the absorption of calcium. OBJECTIVE: We examined the effect of additional caseinphosphopeptides in milk and fermented milk on acute calcium metabolism by measuring intact PTH (iPTH), ionized calcium (iCa), total calcium (Ca) and phosphate (P) from serum, and 24-hour calcium from urine (U-Ca). METHODS: The study consisted of two separate parts, both applying a double-blind randomized crossover study with two interventions, in nine postmenopausal women. The acute effect on calcium metabolism was analysed by measuring iPTH, iCa, Ca and P from serum during the first six hours after the administration of the study milks. U-Ca was analysed 24 hours prior to the study and 0, 2, 4, 6, 12 and 24 hours after the administration of the study milks. The study included two parts, both consisting of two study days with a one-week washout period in between. In the first part the effect of control milk and CPP-enriched milk was measured. The second part evaluated the effect of fermentation by giving subjects milk or fermented milk, both enriched with CPPs. RESULTS: In the first part of the study there were no statistically significant differences in iPTH, iCa, Ca, P or U-Ca between the groups receiving control milk compared to CPP-containing milk. There was no difference in the AUC((0-6)) of iCa and iPTH. In the second part, fermentation did not affect calcium metabolism, when results from the CPP-enriched milk and CPP-enriched fermented milk groups were compared. CONCLUSION: One gram of caseinphosphopeptides does not affect calcium metabolism acutely in postmenopausal women. PMID- 12569120 TI - How does calorie restriction work? PMID- 12569121 TI - The bloody fate of endothelial stem cells. PMID- 12569122 TI - Early-replicating heterochromatin. AB - Euchromatin, which has an open structure and is frequently transcribed, tends to replicate in early S phase. Heterochromatin, which is more condensed and rarely transcribed, usually replicates in late S phase. Here, we report significant deviation from this correlation in the fission yeast, Schizosaccharomyces pombe. We found that heterochromatic centromeres and silent mating-type cassettes replicate in early S phase. Only heterochromatic telomeres replicate in late S phase. Research in other laboratories has shown that occasionally other organisms also replicate some of their heterochromatin in early S phase. Thus, late replication is not an obligatory feature of heterochromatin. PMID- 12569123 TI - The ch-TOG/XMAP215 protein is essential for spindle pole organization in human somatic cells. AB - The ch-TOG/XMAP215 family of proteins bind directly to microtubules and appear to play an essential role in stabilizing spindle microtubules. These proteins stabilize microtubules mainly by influencing microtubule plus-end dynamics, yet, in vivo, they are all strongly concentrated at spindle poles, where the minus ends of the microtubules are concentrated. In Drosophila embryos, the centrosomal protein D-TACC is required to efficiently recruit ch-TOG/Msps to centrosomes. In humans, ch-TOG and the three known TACC proteins have been implicated in cancer, but their functions are unknown. Here we extensively depleted TACC3 and ch-TOG from HeLa cells using RNA interference. In TACC3-depleted cells, spindles are well organized, but microtubules are partially destabilized and ch-TOG is no longer concentrated on spindle microtubules. In ch-TOG-depleted cells, relatively robust spindles form, but the spindles are highly disorganized. Thus, in human somatic cells, ch-TOG appears to play a major role in organizing spindle poles, and a more minor role in stabilizing spindle microtubules that is, at least in part, mediated via an interaction with TACC3. PMID- 12569124 TI - Feedback control of mammalian Hedgehog signaling by the Hedgehog-binding protein, Hip1, modulates Fgf signaling during branching morphogenesis of the lung. AB - Hedgehog (Hh) signaling plays a major role in multiple aspects of embryonic development. A key issue is how negative regulation of Hh signaling might contribute to generating differential responses over tens of cell diameters. In cells that respond to Hh, two proteins that are up-regulated are Patched1 (Ptch1), the Hh receptor, a general target in both invertebrate and vertebrate organisms, and Hip1, a Hh-binding protein that is vertebrate specific. To address the developmental role of Hip1 in the context of Hh signaling, we generated Hip1 mutants in the mouse. Loss of Hip1 function results in specific defects in two Hh target issues, the lung, a target of Sonic hedgehog (Shh) signaling, and the endochondral skeleton, a target of Indian hedgehog (Ihh) signaling. Hh signaling was up-regulated in Hip1 mutants, substantiating Hip1's general role in negatively regulating Hh signaling. Our studies focused on Hip1 in the lung. Here, a dynamic interaction between Hh and fibroblast growth factor (Fgf) signaling, modulated at least in part by Hip1, controls early lung branching. PMID- 12569125 TI - A Serrate-expressing signaling center controls Drosophila hematopoiesis. AB - The differentiation of Drosophila blood cells relies on a functional hierarchy between the GATA protein, Serpent (Srp), and multiple lineage-specific transcription factors, such as the AML1-like protein, Lozenge (Lz). Two major branches of Drosophila hematopoiesis give rise to plasmatocytes/macrophages and crystal cells. Serrate signaling through the Notch pathway is critical in the regulation of Lz expression and the specification of crystal cell precursors, thus providing a key distinction between the two lineages. The expression of Serrate marks a discrete cluster of cells in the lymph gland, a signaling center, with functional similarities to stromal signaling in mammalian hematopoiesis. PMID- 12569126 TI - SCARECROW is involved in positioning the stem cell niche in the Arabidopsis root meristem. AB - Stem cells self-renew and produce daughter cells that differentiate. How stem cells are specified and maintained is a central question in developmental biology. Plant stem cells occupy a small region or niche in larger zones of mitotic activity called meristems. Here we provide molecular evidence that in the Arabidopsis root meristem, the stem cell population depends on a central group of cells, the quiescent center (QC), which positions the stem cell niche. We show that the putative transcription factor SCARECROW (SCR), first identified by its role in radial patterning, is required cell-autonomously for distal specification of the QC, which in turn regulates stem cell fate of immediately surrounding cells. PMID- 12569127 TI - CIAP1 and the serine protease HTRA2 are involved in a novel p53-dependent apoptosis pathway in mammals. AB - Recently a Drosophila p53 protein has been identified that mediates apoptosis via a novel pathway involving the activation of the Reaper gene and subsequent inhibition of the inhibitors of apoptosis (IAPs). The present study found that CIAP1, a major mammalian homolog of Drosophila IAPs, is irreversibly inhibited (cleaved) during p53-dependent apoptosis and this cleavage is mediated by a serine protease. Serine protease inhibitors that block CIAP1 cleavage inhibit p53 dependent apoptosis. Furthermore, activation of the p53 protein increases the transcription of the HTRA2 gene, which encodes a serine protease that interacts with CIAP1 and potentiates apoptosis. These results demonstrate that the mammalian p53 protein may activate apoptosis through a novel pathway functionally similar to that in Drosophila, which involves HTRA2 and subsequent inhibition of CIAP1 by cleavage. PMID- 12569128 TI - Six3 repression of Wnt signaling in the anterior neuroectoderm is essential for vertebrate forebrain development. AB - In vertebrate embryos, formation of anterior neural structures requires suppression of Wnt signals emanating from the paraxial mesoderm and midbrain territory. In Six3(-/-) mice, the prosencephalon was severely truncated, and the expression of Wnt1 was rostrally expanded, a finding that indicates that the mutant head was posteriorized. Ectopic expression of Six3 in chick and fish embryos, together with the use of in vivo and in vitro DNA-binding assays, allowed us to determine that Six3 is a direct negative regulator of Wnt1 expression. These results, together with those of phenotypic rescue of headless/tcf3 zebrafish mutants by mouse Six3, demonstrate that regionalization of the vertebrate forebrain involves repression of Wnt1 expression by Six3 within the anterior neuroectoderm. Furthermore, these results support the hypothesis that a Wnt signal gradient specifies posterior fates in the anterior neural plate. PMID- 12569129 TI - Combinatorial effects of Flk1 and Tal1 on vascular and hematopoietic development in the mouse. AB - Mouse embryos mutant for the VEGF receptor, VEGFR2, Flk-1, or Kdr, fail to form both endothelial and hematopoietic cells, suggesting a possible role in a common progenitor to both lineages. The transcription factor Tal1 (Scl), is not expressed in Flk1(-/-) embryos, consistent with a downstream role in the Flk1 pathway. We tested whether expression of Tal1 under the Flk1 promoter was sufficient to rescue the loss of endothelial and hematopoietic cells in Flk1 mutants. Only partial rescue of hematopoiesis and endothelial development was observed in vivo. However, Flk1(-/Tal1) embryonic stem (ES) cells were capable of blast colony formation in vitro at levels equivalent to Flk1(+/-) heterozygotes. Ectopic expression of Tal1 under the Flk1 promoter in Flk1(+/-) mouse embryos or ES cells caused no obvious pathology but increased the number of blast colony forming cells (BL-CFCs) and enhanced their hematopoietic potential. These single cell-derived BL-CFCs also produced smooth muscle cells in vitro. Increased Tal1 expression inhibited smooth muscle differentiation in this assay, whereas loss of Tal1 promoted smooth muscle formation. We propose a model in which the combinatorial effects of Flk1 and Tal1 act to regulate cell fate choice in early development into hematopoietic, endothelial, and smooth muscle lineages. PMID- 12569131 TI - AFP is a novel negative regulator of ABA signaling that promotes ABI5 protein degradation. AB - Plants have evolved protective mechanisms to ensure their survival when threatened by adverse environmental conditions during their transition to autotrophic growth. During germination, there is a 2- to 3-d period during which a plant can execute growth arrest when challenged by water deficit. This postgermination developmental checkpoint is signaled by the stress hormone abscisic acid (ABA), which induces the expression of the bZIP transcription activator ABI5. The growth arrest efficiency depends on ABI5 levels, and abi5 mutants are ABA-insensitive and unable to execute the ABA-mediated growth arrest. Here we show that a novel ABI5-interacting protein, designated as AFP, can form high molecular weight (Mr) complexes with ABI5 in embryo-derived extracts. Like ABI5, ABI five binding protein (AFP) mRNA and protein levels are induced by ABA during seed germination. Two different afp mutant alleles (afp-1 and afp-2) are hypersensitive to ABA, whereas transgenic plants overexpressing AFP are resistant; in these plants, AFP and ABI5 protein levels are inversely correlated. Genetic analysis shows that abi5-4 is epistatic to afp-1, indicating the ABA hypersensitivity of afp mutants requires ABI5. Proteasome inhibitor studies show that ABI5 stability is regulated by ABA through ubiquitin-related events. When expressed together, AFP and ABI5 are colocalized in nuclear bodies, which also contain COP1, a RING motif protein. Our results suggest that AFP attenuates ABA signals by targeting ABI5 for ubiquitin-mediated degradation in nuclear bodies. PMID- 12569130 TI - Ectodermal Wnt3/beta-catenin signaling is required for the establishment and maintenance of the apical ectodermal ridge. AB - The formation of the apical ectodermal ridge (AER) is critical for the distal outgrowth and patterning of the vertebrate limb. Recent work in the chick has demonstrated that interplay between the Wnt and Fgf signaling pathways is essential in the limb mesenchyme and ectoderm in the establishment and perhaps the maintenance of the AER. In the mouse, whereas a role for Fgfs for AER establishment and function has been clearly demonstrated, the role of Wnt/beta catenin signaling, although known to be important, is obscure. In this study, we demonstrate that Wnt3, which is expressed ubiquitously throughout the limb ectoderm, is essential for normal limb development and plays a critical role in the establishment of the AER. We also show that the conditional removal of beta catenin in the ventral ectodermal cells is sufficient to elicit the mutant limb phenotype. In addition, removing beta-catenin after the induction of the ridge results in the disappearance of the AER, demonstrating the requirement for continued beta-catenin signaling for the maintenance of this structure. Finally, we demonstrate that Wnt/beta-catenin signaling lies upstream of the Bmp signaling pathway in establishment of the AER and regulation of the dorsoventral polarity of the limb. PMID- 12569132 TI - Predicting success in cancer prevention trials. PMID- 12569133 TI - Familial pancreatic cancer: where are we in 2003? PMID- 12569134 TI - Trials examine intraepithelial neoplasia as a marker for cancer. PMID- 12569135 TI - Stat bite: Number of people under age 65 without health insurance, 2002. PMID- 12569136 TI - New carcinogen list includes estrogen, UV radiation. PMID- 12569137 TI - Serving up specimens: NASA-NCI project links databases across the country. PMID- 12569138 TI - Animal models offer insights into human brain tumors. PMID- 12569139 TI - ODAC panel gives nod to Bexxar. PMID- 12569140 TI - Adjuvant breast cancer treatment and cognitive function: current knowledge and research directions. AB - Evidence is mounting that potentially curative systemic adjuvant therapy for early-stage breast cancer may result in cognitive impairment. Five published studies have investigated cognitive function in this setting, and the consistent results of all five studies suggest an adverse effect of adjuvant chemotherapy. These studies are reviewed with particular attention to their methodologic limitations. For example, all five studies used cross-sectional designs, none controlled for possible confounding hormonal factors, and three examined patients who had not received a uniform chemotherapy regimen. The potential roles of chemotherapy-induced menopause and of adjuvant hormonal therapy in cognitive impairment are also discussed. Priorities for future research include confirmation of an effect of adjuvant chemotherapy in a study with a longitudinal design, closer examination of the potential contribution of hormonal factors, and similar studies on the effect of adjuvant therapy on cognitive function in other cancer types. If an effect of systemic adjuvant therapy on cognitive function is confirmed, such an effect will have implications for informed consent. It may also result in incorporation of objective measures of cognition in clinical trials of adjuvant therapy and in the investigation of preventive interventions that might minimize the impact of cognitive dysfunction after cancer treatment. PMID- 12569141 TI - Cyclin D1 genotype, response to biochemoprevention, and progression rate to upper aerodigestive tract cancer. AB - BACKGROUND: Altered cyclin D1 expression in advanced preinvasive lesions of the upper aerodigestive tract (UADT) is associated with an increased risk of developing cancer and histologic progression during and after combination biochemopreventive therapy (13-cis-retinoic acid, alpha-interferon, and alpha tocopherol). Both alleles of the adenine (A)/guanine (G) cyclin D1 polymorphism located at nucleotide 870 encode two alternatively spliced transcripts, but the A allele preferentially encodes a protein with an extended half-life. We investigated whether the cyclin D1 genotype at nucleotide 870 was associated with baseline levels of cyclin D1 protein, post-treatment modulation of cyclin D1 protein levels, histologic response to treatment, and the outcome for subjects with preinvasive UADT lesions after biochemopreventive therapy. METHODS: UADT tissue biopsy samples were obtained before and 6 and 12 months after biochemopreventive treatment from 31 individuals with advanced preinvasive UADT lesions. Tissues were examined for cyclin D1 genotype (by DNA single-strand conformation polymorphism analysis), for cyclin D1 protein expression (by immunohistochemistry), and for cyclin D1 gene copy number (by fluorescence in situ hybridization). Associations of cyclin D1 genotype with histologic response to therapy and time to progression to a higher degree of dysplasia or invasive cancer were investigated. All statistical tests were two-sided. RESULTS: The A allele was associated with increased baseline cyclin D1 expression in the parabasal epithelial layer (16 of 18 AA/AG subjects versus four of nine GG subjects; P =.02), decreased histologic response to biochemopreventive treatment (six of 21 AA/AG subjects versus four of 10 GG subjects; P =.70), decreased favorable modulation of cyclin D1 expression by the treatment (seven of 18 AA/AG subjects versus eight of nine GG subjects; P =.02), and shorter progression-free survival (P =.05). CONCLUSIONS: The cyclin D1 A allele was associated with a diminished modulation of normal physiologic and treatment-induced decreased expression of cyclin D1, a decreased likelihood of response to biochemopreventive intervention, and an increased rate of progression to cancer development, findings that require validation in a larger cohort. PMID- 12569142 TI - Treatment of former smokers with 9-cis-retinoic acid reverses loss of retinoic acid receptor-beta expression in the bronchial epithelium: results from a randomized placebo-controlled trial. AB - BACKGROUND: Loss of retinoic acid receptor beta (RAR-beta) expression in the bronchial epithelium is considered a biomarker of preneoplasia. Retinoids can restore expression of this receptor and, presumably, halt the progression of carcinogenesis. This study was designed to investigate whether either of two retinoid-based regimens, 9-cis-retinoic acid (RA) or 13-cis-RA plus alpha tocopherol (AT), could reverse RAR-beta expression loss in former smokers after 3 months of treatment. METHODS: Individuals (n = 226) who had smoked at least 20 pack-years and had ceased smoking for at least 12 months were randomly assigned to receive 3 months of daily oral 9-cis-RA (100 mg), 13-cis-RA (1 mg/kg) + AT (1200 IU), or placebo. Bronchoscopy and biopsy at six predetermined sites of the bronchial tree were performed before treatment and at 3 and 6 months thereafter. Specimens were evaluated for squamous metaplasia, dysplasia, and RAR-beta expression. McNemar's test was used to test changes in RAR-beta expression and squamous metaplasia within each treatment group, and a generalized estimating equations model was applied to model the treatment effect, adjusting for covariates. All statistical tests were two-sided. RESULTS: A total of 177 assessable subjects completed at least 3 months of therapy and underwent at least the baseline and 3-month bronchoscopic evaluations with biopsies. RAR-beta was detected in 69.7% of all baseline biopsy samples, and metaplasia was evident in 6.9% of all baseline samples from 240 subjects. Restoration of RAR-beta expression (P =.03) and reduction of metaplasia (P =.01) were found in the 9-cis RA group. After adjustment for years of smoking, packs/day smoked, and metaplasia, treatment with 9-cis-RA, but not with 13-cis-RA + AT, led to a statistically significant increase in RAR-beta expression compared with placebo (P =.03). CONCLUSION: 9-cis-RA treatment can restore RAR-beta expression in the bronchial epithelium of former smokers, raising the possibility that this retinoid has potential chemopreventive properties in former smokers. PMID- 12569143 TI - BRCA2 germline mutations in familial pancreatic carcinoma. AB - BACKGROUND: Although as many as 10% of pancreatic cancer cases may have an inherited component, familial pancreatic cancer has not been linked to defects in any specific gene. Some studies have shown that families with germline mutations in the breast cancer susceptibility gene BRCA2 have an increased risk of breast and ovarian cancers, as well as a modestly increased risk of pancreatic cancer. To study these relationships in more detail, we examined whether BRCA2 germline mutations are associated with familial pancreatic cancer. METHODS: We identified 26 European families in which at least two first-degree relatives had a histologically confirmed diagnosis of pancreatic ductal adenocarcinoma. We sequenced genomic DNA isolated from peripheral blood lymphocytes obtained from participating family members to identify germline mutations in BRCA2. RESULTS: Three (12%, exact 95% confidence interval [CI] = 2% to 30%) families carried germline frameshift mutations in the BRCA2 gene that are predicted to result in a truncated BRCA2 protein. Two additional families harbored mutations previously designated as unclassified variants of BRCA2. Thus, 19% (exact 95% CI = 7% to 39%) of the families in our study had either a frameshift mutation or an unclassified variant of BRCA2. None of the families in our study met the criteria for familial breast or ovarian cancer. CONCLUSIONS: Our data support an important role for BRCA2 germline mutations in a subpopulation of families with familial pancreatic cancer. BRCA2 mutation analysis should be included in molecular genetic testing and counseling strategies in families with at least two first degree relatives affected with ductal adenocarcinoma of the pancreas. PMID- 12569144 TI - Single- versus multiple-fraction radiotherapy in patients with painful bone metastases: cost-utility analysis based on a randomized trial. AB - BACKGROUND: Radiotherapy is an effective palliative treatment for cancer patients with painful bone metastases. Although single- and multiple-fraction radiotherapy are thought to provide equal palliation, which treatment schedule provides better value for the money is unknown. We compared quality-adjusted life expectancy (the overall valuation of the health of the patients) and societal costs for patients receiving either single- or multiple-fraction radiotherapy. METHODS: A societal cost-utility analysis was performed on a Dutch randomized, controlled trial of 1157 patients with painful bone metastases that compared pain responses and quality of life from a single-fraction treatment schedule of 8 Gy with a treatment schedule of six fractions of 4 Gy each. The societal values of life expectancies were assessed with the EuroQol classification system (EQ-5D) questionnaire. A subset of 166 patients also answered additional questionnaires to estimate nonradiotherapy and nonmedical costs. Statistical tests were two sided. RESULTS: Comparing the single- and multiple-fraction radiotherapy schedules, no differences were found in life expectancy (43.0 versus 40.4 weeks, P =.20) or quality-adjusted life expectancy (17.7 versus 16.0 weeks, P =.21). The estimated cost of radiotherapy, including retreatments and nonmedical costs, was statistically significantly lower for the single-fraction schedule than for the multiple-fraction schedule ($2438 versus $3311, difference = $873, 95% confidence interval [CI] on the difference = $449 to $1297; P<.001). The estimated difference in total societal costs was larger, also in favor of the single fraction schedule, but it was not statistically significant ($4700 versus $6453, difference = $1753, 95% CI on the difference = -$99 to $3604; P =.06). For willingness-to-pay between $5000 and $40 000 per quality-adjusted life year, the single-fraction schedule was statistically significantly more cost-effective than the multiple-fraction schedule (P< or =.05). CONCLUSIONS: Compared with multiple fraction radiotherapy, single-fraction radiotherapy provides equal palliation and quality of life and has lower medical and societal costs, at least in The Netherlands. Therefore, single-fraction radiotherapy should be considered as the palliative treatment of choice for cancer patients with painful bone metastases. PMID- 12569145 TI - Risk of perforation after colonoscopy and sigmoidoscopy: a population-based study. AB - BACKGROUND: Although the risk of bowel perforation is often cited as a major factor in the choice between colonoscopy and sigmoidoscopy for colorectal screening, good estimates of the absolute and relative risks of perforation are lacking. METHODS: We used a large population-based cohort that consisted of a random sample of 5% of Medicare beneficiaries living in regions of the United States covered by the Surveillance, Epidemiology, and End Results (SEER) Program registries to determine rates of perforation in people aged 65 years and older. We identified individuals who were cancer-free and had undergone colonoscopy or sigmoidoscopy between 1991 and 1998, calculated both the incidence and risk of perforation within 7 days of the procedure, and explored the impact on incidence and risk of perforation of age, race/ethnicity, sex, comorbidities, and indication for the procedure. We also estimated the risk of death after perforation. Risks were calculated with odds ratios (ORs) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: There were 77 perforations after 39 286 colonoscopies (incidence = 1.96/1000 procedures) and 31 perforations after 35 298 sigmoidoscopies (incidence = 0.88/1000 procedures). After adjustment, the OR for perforation from colonoscopy relative to perforation from sigmoidoscopy was 1.8 (95% CI = 1.2 to 2.8). Risk of perforation from either procedure increased in association with increasing age (P(trend)<.001 for both procedures) and the presence of two or more comorbidities (P(trend)<.001 for colonoscopy and P(trend) =.03 for sigmoidoscopy). Compared with those who were endoscopied and did not have a perforation, the risk of death was statistically significantly increased for those who had a perforation after either colonoscopy (OR = 9.0, 95% CI = 3.0 to 27.3) or sigmoidoscopy (OR = 8.8, 95% CI = 1.6 to 48.5). The risk of perforation after colonoscopy, especially for screening procedures, declined during the 8-year study period. CONCLUSIONS: The risk of perforation after colonoscopy is approximately double that after sigmoidoscopy, but this difference appears to be decreasing. These observations should be useful to clinicians making screening and diagnostic decisions for individual patients and to policy officials setting guidelines for colorectal cancer screening programs. PMID- 12569146 TI - Effects of dietary selenium supplementation on DNA damage and apoptosis in canine prostate. AB - The trace mineral selenium inhibits cancer development in a variety of experimental animal models. We used an in vivo canine model to evaluate the effects of dietary selenium supplementation on DNA damage in prostate tissue and on apoptosis in prostate epithelial cells. Sexually intact elderly male beagle dogs were randomly assigned to receive an unsupplemented diet (control group) or diets that were supplemented with selenium (treatment group), either as selenomethionine or as high-selenium yeast at 3 micro g/kg or 6 micro g/kg body weight per day for 7 months. The extent of DNA damage in prostate cells and in peripheral blood lymphocytes, as determined by the alkaline comet assay, was lower among the selenium-supplemented dogs than among the control dogs (prostate P<.001; peripheral blood lymphocytes P =.003; analysis of variance) but was not associated with the activity of the antioxidant enzyme glutathione peroxidase in plasma. The median number of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling-positive (i.e., apoptotic) prostate epithelial cells was 3.7 (interquartile range = 1.1-7.6) for the selenium-supplemented dogs and 1.7 (interquartile range = 0.2-2.8) for the control dogs ( P =.04, Mann-Whitney U test). These data suggest that dietary selenium supplementation decreases DNA damage and increases epithelial cell apoptosis within the aging canine prostate. PMID- 12569147 TI - Re: Sex-related differences in bronchial epithelial changes associated with tobacco smoking. PMID- 12569150 TI - New insights into the pathogenesis of asthma. PMID- 12569151 TI - Huntingtin in health and disease. PMID- 12569152 TI - Neuronal degeneration and mitochondrial dysfunction. PMID- 12569153 TI - DAP12 signaling: from immune cells to bone modeling and brain myelination. PMID- 12569154 TI - Artemis: guarding small children and, now, the genome. PMID- 12569155 TI - HIV protease inhibitors and atherosclerosis. PMID- 12569156 TI - Endothelial lipase: direct evidence for a role in HDL metabolism. PMID- 12569157 TI - Osteopetrosis and thalamic hypomyelinosis with synaptic degeneration in DAP12 deficient mice. AB - Deletions in the DAP12 gene in humans result in Nasu-Hakola disease, characterized by a combination of bone fractures and psychotic symptoms similar to schizophrenia, rapidly progressing to presenile dementia. However, it is not known why these disorders develop upon deficiency in DAP12, an immunoreceptor signal activator protein initially identified in the immune system. Here we show that DAP12-deficient (DAP12(-/-)) mice develop an increased bone mass (osteopetrosis) and a reduction of myelin (hypomyelinosis) accentuated in the thalamus. In vitro osteoclast induction from DAP12(-/-) bone marrow cells yielded immature cells with attenuated bone resorption activity. Moreover, immature oligodendrocytes were arrested in the vicinity of the thalamus, suggesting that the primary defects in DAP12(-/-) mice are the developmental arrest of osteoclasts and oligodendrocytes. In addition, the mutant mice also showed synaptic degeneration, impaired prepulse inhibition, which is commonly observed in several neuropsychiatric diseases in humans including schizophrenia, and aberrant electrophysiological profiles in the thalami. These results provide a molecular basis for a unique combination of skeletal and psychotic characteristics of Nasu-Hakola disease as well as for schizophrenia and presenile dementia. PMID- 12569158 TI - Decreased atherosclerosis in CX3CR1-/- mice reveals a role for fractalkine in atherogenesis. AB - The hallmark of early atherosclerosis is the accumulation of lipid-laden macrophages in the subendothelial space. Circulating monocytes are the precursors of these "foam cells," and recent evidence suggests that chemokines play important roles in directing monocyte migration from the blood to the vessel wall. Fractalkine (FK) is a structurally unusual chemokine that can act either as a soluble chemotactic factor or as a transmembrane-anchored adhesion receptor for circulating leukocytes. A polymorphism in the FK receptor, CX(3)CR1, has been linked to a decrease in the incidence of coronary artery disease. To determine whether FK is critically involved in atherogenesis, we deleted the gene for CX(3)CR1 and crossed these mice into the apoE(-/-) background. Here we report that FK is robustly expressed in lesional smooth muscle cells, but not macrophages, in apoE(-/-) mice on a high-fat diet. CX(3)CR1(-/-) mice have a significant reduction in macrophage recruitment to the vessel wall and decreased atherosclerotic lesion formation. Taken together, these data provide strong evidence that FK plays a key role in atherogenesis. PMID- 12569159 TI - A common SCN5A polymorphism modulates the biophysical effects of an SCN5A mutation. AB - Our understanding of the genetic basis of disease has expanded with the identification of rare DNA sequence variations ("mutations") that evoke inherited syndromes such as cystic fibrosis, congenital epilepsy, and cardiac arrhythmias. Common sequence variants ("polymorphisms") have also been implicated as risk factors in multiple diseases. Mutations in SCN5A, the cardiac Na(+) channel gene, that cause a reduction in Na(+) current may evoke severe, life-threatening disturbances in cardiac rhythm (i.e., Brugada syndrome), isolated cardiac conduction disease, or combinations of these disorders. Conduction disease is manifest clinically as heart rate slowing (bradycardia), syncope, or "lightheadedness". Recent electrophysiologic studies reveal that mutations in particular families exhibiting cardiac conduction disease cause marked effects on several competing voltage-dependent gating processes, but nonetheless cause a mild "net" reduction in Na(+) current. Here we show that a common SCN5A polymorphism (H558R) in the Na(+) channel I-II interdomain cytoplasmic linker, present in 20% of the population, can mitigate the in vitro effects of a nearby mutation (T512I) on Na(+) channel function. The mutation and the polymorphism were both found in the same allele of a child with isolated conduction disease, suggesting a direct functional association between a polymorphism and a mutation in the same gene. PMID- 12569160 TI - Endothelial lipase is a major determinant of HDL level. AB - A new member of the lipase gene family, initially termed endothelial lipase (gene nomenclature, LIPG; protein, EL), is expressed in a variety of different tissues, suggesting a general role in lipid metabolism. To assess the hypothesis that EL plays a physiological role in lipoprotein metabolism in vivo, we have used gene targeting of the native murine locus and transgenic introduction of the human LIPG locus in mice to modulate the level of EL expression. Evaluation of these alleles in a C57Bl/6 background revealed an inverse relationship between HDL cholesterol level and EL expression. Fasting plasma HDL cholesterol was increased by 57% in LIPG(-/-) mice and 25% in LIPG(+/-) mice and was decreased by 19% in LIPG transgenic mice as compared with syngeneic controls. Detailed analysis of lipoprotein particle composition indicated that this increase was due primarily to an increased number of HDL particles. Phospholipase assays indicated that EL is a primary contributor to phospholipase activity in mouse. These data indicate that expression levels of this novel lipase have a significant effect on lipoprotein metabolism. PMID- 12569161 TI - Inhibition of endothelial lipase causes increased HDL cholesterol levels in vivo. AB - Endothelial lipase (EL) is a recently discovered member of the lipoprotein lipase gene family that hydrolyzes HDL phospholipids ex vivo and reduces HDL cholesterol (HDL-C) levels when overexpressed in vivo in mice. To gain further insight into the physiological role of EL in the metabolism of HDL in vivo, studies were performed in which EL was inhibited in wild-type, hepatic lipase knockout (HL(-/ )), and human apoA-I transgenic mice by intravenous infusion of a polyclonal antibody inhibitory to murine EL. As compared with infusion of a control antibody, infusion of the inhibitory antibody resulted in a 25-60% increase in HDL-C levels in the three mouse models, with the peak HDL-C levels occurring at 48 hours after injection. Inhibition of EL also generated larger HDL particles in the HL(-/-) mice. The clearance of HDL phospholipid was significantly slower in human apoA-I transgenic mice injected with an antibody against murine EL (mEL) than in mice injected with a control antibody. We conclude that inhibition of EL results in increased HDL-C levels and that EL is an important enzyme in the physiological regulation of HDL metabolism. PMID- 12569162 TI - Costimulating aberrant T cell responses by B7-H1 autoantibodies in rheumatoid arthritis. AB - A pathogenic hallmark of rheumatoid arthritis (RA) is persistent activation of self-reactive CD4(+) T cells. The cause of this aberrant activity remains elusive. We report here detection of autoantibodies against B7-H1, a recently described member of the B7 family, in 29% of patients with RA versus 4% of healthy donors. High-level expression of cell surface B7-H1 are found on activated human CD4(+), CD8(+), and CD45RO(+) T cells. Immobilized autoantibodies to B7-H1 are capable of costimulating the proliferation of CD4(+) T cells in vitro, and the presence of these autoantibodies correlates with active disease status. Using immobilized B7-H1 mAb's and programmed death 1Ig, we demonstrate that engagement of B7-H1 on CD4(+) T cells costimulates proliferation and secretion of IL-10, and subsequently leads to programmed cell death, accompanied with upregulated expression of TNF-related apoptosis-inducing ligand and activation of caspase-3. Taken together with our previous findings, these data indicate a bidirectional signaling role of B7-H1 in T cell costimulation and apoptosis and implicate B7-H1 autoantibodies as contributing to the progression of RA by inducing aberrant T cell responses. PMID- 12569163 TI - P2Y6 receptor mediates colonic NaCl secretion via differential activation of cAMP mediated transport. AB - Extracellular nucleotides are important regulators of epithelial ion transport. Here we investigated nucleotide-mediated effects on colonic NaCl secretion and the signal transduction mechanisms involved. Basolateral UDP induced a sustained activation of Cl(-) secretion, which was completely inhibited by 293B, a specific inhibitor of cAMP-stimulated basolateral KCNQ1/KCNE3 K(+) channels. We therefore speculated that a basolateral P2Y(6) receptor could increase cAMP. Indeed UDP elevated cAMP in isolated crypts. We identified an epithelial P2Y(6) receptor using crypt [Ca(2+)](i) measurements, RT-PCR, and immunohistochemistry. To investigate whether the rat P2Y(6)elevates cAMP, we coexpressed the P2Y(1) or P2Y(6) receptor together with the cAMP-regulated cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel in Xenopus oocytes. A two-electrode voltage clamp was used to monitor nucleotide-induced Cl(-) currents. In oocytes expressing the P2Y(1) receptor, ATP transiently activated the endogenous Ca(2+) activated Cl(-) current, but not CFTR. In contrast, in oocytes expressing the P2Y(6)receptor, UDP transiently activated the Ca(2+)-activated Cl(-) current and subsequently CFTR. CFTR Cl(-) currents were identified by their halide conductance sequence. In summary we find a basolateral P2Y(6) receptor in colonic epithelial cells stimulating sustained NaCl secretion by way of a synergistic increase of [Ca(2+)](i) and cAMP. In support of these data P2Y(6) receptor stimulation differentially activates CFTR in Xenopus oocytes. PMID- 12569164 TI - Partial T and B lymphocyte immunodeficiency and predisposition to lymphoma in patients with hypomorphic mutations in Artemis. AB - We have previously described the identification of Artemis, a factor involved in the nonhomologous end joining (NHEJ) phase of V(D)J recombination of T and B cell receptor genes. Null mutations of the Artemis gene result in a complete absence of T and B lymphocytes that is associated with increased cell radiosensitivity, causing the radiosensitive T(-)B(-) SCID (RS-SCID) condition. We presently report the occurrence of hypomorphic mutations of the Artemis gene in four patients from two kindreds. Partially preserved in vivo activity of Artemis is associated with the presence of polyclonal T and B lymphocyte populations, albeit in reduced numbers, along with chromosomal instability and development of EBV-associated lymphoma in two of four patients. This syndrome emphasizes the role of Artemis in the NHEJ pathway of DNA repair and suggests that other, yet ill-defined, conditions associating immunodeficiency and lymphoma could be caused by mutations in genes encoding NHEJ factors. PMID- 12569165 TI - HIV protease inhibitors promote atherosclerotic lesion formation independent of dyslipidemia by increasing CD36-dependent cholesteryl ester accumulation in macrophages. AB - Protease inhibitors decrease the viral load in HIV patients, however the patients develop hypertriglyceridemia, hypercholesterolemia, and atherosclerosis. It has been assumed that protease inhibitor-dependent increases in atherosclerosis are secondary to the dyslipidemia. Incubation of THP-1 cells or human PBMCs with protease inhibitors caused upregulation of CD36 and the accumulation of cholesteryl esters. The use of CD36-blocking antibodies, a CD36 morpholino, and monocytes isolated from CD36 null mice demonstrated that protease inhibitor induced increases in cholesteryl esters were dependent on CD36 upregulation. These data led to the hypothesis that protease inhibitors induce foam cell formation and consequently atherosclerosis by upregulating CD36 and cholesteryl ester accumulation independent of dyslipidemia. Studies with LDL receptor null mice demonstrated that low doses of protease inhibitors induce an increase in the level of CD36 and cholesteryl ester in peritoneal macrophages and the development of atherosclerosis without altering plasma lipids. Furthermore, the lack of CD36 protected the animals from protease inhibitor-induced atherosclerosis. Finally, ritonavir increased PPAR-gamma and CD36 mRNA levels in a PKC- and PPAR-gamma dependent manner. We conclude that protease inhibitors contribute to the formation of atherosclerosis by promoting the upregulation of CD36 and the subsequent accumulation of sterol in macrophages. PMID- 12569166 TI - Paradoxical resistance to diet-induced obesity in UCP1-deficient mice. AB - The availability of mice lacking the mitochondrial uncoupling protein UCP1, has provided an opportunity to analyze the relationship between the capacity for energy expenditure and the development of obesity in response to a high-fat, high sucrose diet. Congenic UCP1-deficient mice on a C57BL/6J genetic background show a temperature-dependent resistance to diet-induced obesity when compared with wild-type mice. This resistance, which occurs at 20 degrees C, is quickly reversed when the ambient temperature is increased to 27 degrees C. At 20 degrees C, total oxygen consumption and physical activity of mutant and wild-type mice are indistinguishable; however, body temperature is higher in UCP1-deficient mice by 0.1-0.3 degrees C, and respiratory quotient is slightly reduced. A reduced respiratory quotient, together with elevated beta-hydroxybutyrate and reduced plasma fatty acid levels, suggests that the mutants oxidize a greater proportion of fat than wild-type mice, and that this possibly accounts for the resistance to diet-induced obesity. Although shivering is one alternative mechanism of thermogenesis that is probably used in UCP1-deficient mice, whether there are others remains to be determined. Nevertheless, our study underscores the paradox that elimination of the major thermogenic mechanism in the animal reduces rather than increases metabolic efficiency. We propose that in the absence of nonshivering thermogenesis, alternative, calorically more costly pathways of metabolism must be used to maintain body temperature. PMID- 12569167 TI - Activation of human T cells by FcR nonbinding anti-CD3 mAb, hOKT3gamma1(Ala-Ala). AB - Dimeric Fc receptor (FcR) nonbinding anti-CD3 antibodies have been developed to minimize toxicities associated with classical anti-CD3 monoclonal antibodies (e.g., OKT3). Studies with murine analogs of non-FcR-binding antibodies have shown reduced mitogenicity compared to OKT3. In a trial of an FcR nonbinding humanized anti-CD3 mAb hOKT3gamma1(Ala-Ala) for treatment of patients with type 1 diabetes, we found significant increases in IL-10 and IL-5 in the serum of 63% and 72% of patients, respectively, and TNF-alpha and IL-6 levels that were lower than those previously reported following OKT3 therapy. The activation signal delivered by hOKT3gamma1(Ala-Ala) was associated with calcium signaling and cytokine production by previously activated human cells in vitro. However, the production of IL-10, compared to IFN-gamma on a molar basis, was greater after culture with hOKT3gamma1(Ala-Ala) than with OKT3. Flow cytometric studies confirmed that OKT3 induced IFN-gamma and IL-10 production, but hOKT3gamma1(Ala Ala) induced only detectable IL-10 production in CD45RO(+) cells. Moreover, in vivo, we found IL-10(+)CD4(+) T cells after drug treatment. These cells were heterogeneous but generally CD45RO(+), CTLA-4(-), and expressed CCR4. A subgroup of these cells expressed TGF-beta. Thus, the non-FcR binding anti-CD3 mAb, hOKT3gamma1(Ala-Ala) delivers an activation signal to T cells that is quantitatively and qualitatively different from OKT3. It leads to the generation of T cells that might inhibit the autoimmune response and may be involved in the beneficial effect on beta cell destruction in Type 1 diabetes. PMID- 12569168 TI - Lipoprotein lipase (LpL) on the surface of cardiomyocytes increases lipid uptake and produces a cardiomyopathy. AB - Lipoprotein lipase is the principal enzyme that hydrolyzes circulating triglycerides and liberates free fatty acids that can be used as energy by cardiac muscle. Although lipoprotein lipase is expressed by and is found on the surface of cardiomyocytes, its transfer to the luminal surface of endothelial cells is thought to be required for lipoprotein lipase actions. To study whether nontransferable lipoprotein lipase has physiological actions, we placed an alpha myosin heavy-chain promoter upstream of a human lipoprotein lipase minigene construct with a glycosylphosphatidylinositol anchoring sequence on the carboxyl terminal region. Hearts of transgenic mice expressed the altered lipoprotein lipase, and the protein localized to the surface of cardiomyocytes. Hearts, but not postheparin plasma, of these mice contained human lipoprotein lipase activity. More lipid accumulated in hearts expressing the transgene; the myocytes were enlarged and exhibited abnormal architecture. Hearts of transgenic mice were dilated, and left ventricular systolic function was impaired. Thus, lipoprotein lipase expressed on the surface of cardiomyocytes can increase lipid uptake and produce cardiomyopathy. PMID- 12569169 TI - Gain control of firing rate by shunting inhibition: roles of synaptic noise and dendritic saturation. AB - Adjusting input-output gain is crucial for information processing by the brain. Gain control of subthreshold depolarization is commonly ascribed to increased membrane conductance caused by shunting inhibition. But contrary to its divisive effect on depolarization, shunting inhibition on its own fails to divisively modulate firing rate, apparently upsetting a critical tenet of neural models that use shunting inhibition to achieve gain control. Using a biophysically realistic neuron model, we show that divisive modulation of firing rate by shunting inhibition requires synaptic noise to smooth the relation between firing rate and somatic depolarization; although necessary, noise alone endows shunting inhibition with only a modest divisive effect on firing rate. In addition to introducing noise, synaptic input is associated with a nonlinear relation between somatic depolarization and excitation because of dendritic saturation; this nonlinearity dramatically enhances divisive modulation of firing rate by shunting inhibition under noisy conditions. Thus, shunting inhibition can act as a mechanism for firing rate gain control, but its modulatory effects (which include both divisive and subtractive components) are fully explained only when both synaptic noise and dendritic saturation are taken into account. PMID- 12569170 TI - Tracking an embryonic L1 retrotransposition event. AB - Long interspersed nuclear elements 1 (L1) are active retrotransposons that reside in many species, including humans and rodents. L1 elements produce an RNA intermediate that is reverse transcribed to DNA and inserted in a new genomic location. We have tagged an active human L1 element (L1(RP)) with a gene encoding enhanced GFP (EGFP). Expression of GFP occurs only if L1-EGFP has undergone a cycle of transcription, reverse transcription, and integration into a transcriptionally permissive genomic region. We show here that L1-EGFP can undergo retrotransposition in vivo and produce fluorescence in mouse testis. The retrotransposition event characterized here has occurred at a very early stage in the development of an L1-EGFP transgenic founder mouse. PMID- 12569172 TI - Pretarget radiotherapy with an anti-CD25 antibody-streptavidin fusion protein was effective in therapy of leukemia/lymphoma xenografts. AB - Although radioimmunotherapy with radiolabeled intact monoclonal antibodies has demonstrated efficacy in the treatment of lymphoma, it provides low tumor-to normal-tissue radionuclide target ratios and unwanted prolonged radiation exposure to the bone marrow. To overcome these obstacles, the administration of the radionuclide was separated from that of the antibody by using an anti-IL-2 receptor alpha antibody single chain Fv-streptavidin fusion protein, followed by radiolabeled biotin to treat lymphoma or leukemia xenografted mice. This Pretarget approach provided extremely rapid and effective tumor targeting, permitting the use of short-lived alpha-emitting radionuclides. With the beta emitter (90)Y, all of the 10 lymphoma-xenografted mice were cured. With the alpha emitter (213)Bi, significant efficacy was obtained in treating leukemic mice, and, furthermore, when combined with immunotherapy, 7 of 10 leukemic mice were cured. Thus, Pretarget radioimmunotherapy is very promising and could represent the next generation in the treatment of lymphoma and leukemia. PMID- 12569171 TI - Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection. AB - Streptococcus pneumoniae is one of the leading causes of invasive bacterial disease worldwide. Fragments of the cell wall and the cytolytic toxin pneumolysin have been shown to contribute substantially to inflammatory damage, although the interactions between pneumococcal components and host-cell structures have not been elucidated completely. Results of a previous study indicated that cell-wall components of pneumococci are recognized by Toll-like receptor (TLR)2 but suggested that pneumolysin induces inflammatory events independently of this receptor. In this study we tested the hypothesis that pneumolysin interacts with surface proteins of the TLR family other than TLR2. We found that pneumolysin stimulates tumor necrosis factor-alpha and IL-6 release in wild-type macrophages but not in macrophages from mice with a targeted deletion of the cytoplasmic TLR adapter molecule myeloid differentiation factor 88, suggesting the involvement of the TLRs in pneumolysin recognition. Purified pneumolysin synergistically activated macrophage responses together with preparations of pneumococcal cell walls or staphylococcal peptidoglycan, which are known to activate TLR2. Furthermore, when compared with wild-type macrophages, macrophages from mice that carry a spontaneous mutation in TLR4 (P712H) were hyporesponsive to both pneumolysin alone and the combination of pneumolysin with pneumococcal cell walls. Finally, these TLR4-mutant mice were significantly more susceptible to lethal infection after intranasal colonization with pneumolysin-positive pneumococci than were control mice. We conclude that the interaction of pneumolysin with TLR4 is critically involved in the innate immune response to pneumococcus. PMID- 12569173 TI - Mapping the binding interface between human eukaryotic initiation factors 1A and 5B: a new interaction between old partners. AB - The translation initiation factors (IFs) IF1/eIF1A and IF2e/IF5B have been conserved throughout all kingdoms. Although the central roles of the bacterial factors IF1 and IF2 were established long ago, the importance of their eukaryotic homologs, eukaryotic IFs (eIFs) eIF1A and eIF5B, has only recently become evident. The translation machinery in eukaryotes is more complex and accordingly, eIF1A and eIF5B seem to have acquired a number of new functions while also retaining many of the roles of bacterial IF1 and IF2. IF1 and IF2 have been shown to interact on the ribosome but no binding has been detected for the free factors. In contrast, yeast eIF1A and eIF5B have been reported to interact in the absence of ribosomes. Here, we have identified the binding interface between human eIF1A and the C-terminal domain of eIF5B by using solution NMR. That interaction interface involves the C termini of the two proteins, which are not present in bacterial IF1 and IF2. The interaction is, therefore, unique to eukaryotes. A structural model for the interaction of eIF1A and eIF5B in the context of the ribosome is presented. We propose that eIF1A and eIF5B simultaneously interact at two sites that are >50 A apart: through their C termini as reported here, and through an interface previously identified in bacterial IF1 and IF2. The binding between the C termini of eIF1A and eIF5B has implications for eukaryote-specific mechanisms of recruitment and release of translation IFs from the ribosome. PMID- 12569174 TI - Microsatellite instability and defects in mismatch repair proteins: a new aetiology for Sertoli cell-only syndrome. AB - Microsatellite instability is characteristic of certain types of cancer, and is present in rodents lacking specific DNA mismatch repair proteins. These azoospermic mice exhibit spermatogenic defects similar to some human testicular failure patients. Therefore, we hypothesized that microsatellite instability due to deficiencies in mismatch repair genes might be an unrecognized aetiology of human testicular failure. Because these azoospermic patients are candidates for testicular sperm extraction and ICSI, transmission of mismatch repair defects to the offspring is possible. Seven microsatellite loci were analysed for instability in specimens from 41 testicular failure patients and 20 controls. Blood and testicular DNA were extracted from patient and control specimens, and amplified by PCR targeting seven microsatellite loci. DNA fragment length was analysed with an ABI Prism 310 Genotyping Machine and GeneScan software. Immunohistochemistry was performed on paraffinized testis biopsy sections and cultured testicular fibroblasts from each patient to determine if expression of the mismatch repair proteins hMSH2 and hMLH1 was normal in both somatic and germline cells. Results demonstrate that microsatellite instability and DNA mismatch repair protein defects are present in some azoospermic men, predominantly in Sertoli cell-only patients (P < 0.01 and P < 0.05 respectively). This provides evidence of a previously unrecognized aetiology of testicular failure that may be associated with cancer predisposition. PMID- 12569175 TI - Single nucleotide polymorphisms in the protamine-1 and -2 genes of fertile and infertile human male populations. AB - Although various genetic factors have been implicated in human male infertility, the causative genes for the different types of idiopathic male infertility have not been elucidated. Protamines, which are the major DNA-binding proteins in the sperm nucleus, package the DNA into the sperm head. Analysis of the human protamine-1 (PRM1) and -2 (PRM2) gene sequences in 226 sterile male patients and in 270 proven-fertile male volunteers revealed four single nucleotide polymorphisms (SNPs) in the PRM1 coding region, which did not cause any amino acid substitutions, and one SNP in the PRM2 gene, which produced translation termination. We also observed one SNP in the 3' non-coding region of the PRM1 gene, and two SNPs within the intron of the PRM2 gene. The prevalence of these SNPs was similar in both infertile patients and in proven-fertile volunteers, except that the c248t alteration in the PRM2 gene induced a nonsense codon under conditions of heterozygosity in one infertile patient. Although the PRM1 and PRM2 genes are highly conserved, the single SNP in the PRM2 gene that induces translation termination may result in male infertility due to haploinsufficiency of PRM2. PMID- 12569177 TI - Expression of beta hCG and alpha CG mRNA and hCG hormone in human decidual tissue in patients during tubal pregnancy. AB - We recently showed that endometrial tissue produces hCG during the secretory phase of the menstrual cycle. Based on these findings, we hypothesized that the decidua should also be able to secrete hCG. We examined the decidualized endometrium of patients with extrauterine pregnancies. Decidual specimens were obtained for mRNA extraction and paraffin embedding from 24 patients that were between weeks 6-11 of tubal pregnancy. Tissues were evaluated and classified into one of three groups based on the endometrial differentiation that took place prior to conception: (A) high secretory transformation, (B) diminished transformation with restricted decidualization and (C) inferior endometrial proliferation. Decidual gland hCG secretion was demonstrated immunohistochemically and by Western blotting. Serum hCG levels were higher (P < 0.0001) in patients from group A than group C. mRNA expression of both the beta subunit (beta-hCG) and alpha subunit (alpha-CG) was determined by RT-PCR. Furthermore, the specificity of beta-hCG amplification was confirmed by restriction enzymes. beta-LH amplification was not found. Moreover, the degree of endometrial transformation and the level of decidualization was found to correlate with hCG hormone staining and beta-hCG mRNA expression. hCG protein in the decidua was present in the glands of the compact layer and in the spongy layer, and was more pronounced in previously transformed high secretory endometrium than in inferior endometrium. In conclusion, this study provides the first evidence that hCG is produced in the decidua of patients during extrauterine pregnancies and might play a possible paracrine role. PMID- 12569176 TI - Expression and function of interleukin-11 and its receptor alpha in the human endometrium. AB - The interleukin-11 (IL-11) receptor alpha has an important function in decidualization of mouse endometrial stroma but the function of IL-11 and its receptor in the human endometrium remains unknown. The mRNA for IL-11 and its receptor alpha in human endometrial tissue samples were analysed by semi quantitative RT-PCR and RNase protection assays respectively. The proteins were detected in frozen endometrial tissue samples by immunofluorescence. The effect of heparin-binding epidermal growth factor (HB-EGF) on secretion of IL-11 by cultured endometrial stromal cells was assessed by enzyme-linked immunosorbent assay. The proliferative potential of IL-11 in endometrial stromal cells was assessed by [(3)H]thymidine uptake. IL-11 and its receptor alpha mRNAs and proteins were detected in the endometrium throughout the cycle. Distinct patterns of localization of the ligand and receptor were observed. HB-EGF induced IL-11 secretion by cultured stromal cells, and IL-11 induced [(3)H]thymidine uptake by these cells. Our data suggest that IL-11-receptor interactions may perform different functions in the human endometrium at different stages of the cycle, and that secretion of IL-11 is modulated by local growth factors. PMID- 12569178 TI - Transforming growth factor beta1 exerts an autocrine regulatory effect on human endometrial stromal cell apoptosis, involving the FasL and Bcl-2 apoptotic pathways. AB - Transforming growth factor beta1 (TGFbeta1) is expressed in human endometrium. It regulates epithelial cell proliferation and apoptosis. The aim of the present work was to examine the role of TGFbeta1 on human endometrial stromal cell apoptosis. Primary cultures of isolated stromal cells were obtained from biopsies of late secretory phase endometrium. We have found the following: (i) TGFbeta1 induced apoptosis of stromal cells in a time- and dose-dependent manner; (ii) blockade of TGFbeta1's autocrine/paracrine effect by TGFbeta1-neutralizing antibodies diminished the basal rate of stromal cell apoptosis; (iii) semi quantitative Western blot analysis showed that TGFbeta1 caused a rapid but transient elevation of the pro-apoptotic FasL protein, without affecting the levels of Fas receptor; (iv) TGFbeta1 increased the levels of the anti-apoptotic Bcl-2 and Bcl-xL proteins, while having no significant effects on the pro apoptotic proteins Bax and Bak, suggesting the activation of a transient survival mechanism activated in stromal cells as a parallel rescue response to the apoptosis-inducing FasL protein. In conclusion, our data provide evidence that TGFbeta1 exerts an autocrine pro-apoptotic effect on human endometrial stroma, via the FasL/Fas system. PMID- 12569179 TI - Fractalkine (FRK) levels in amniotic fluid and its production during pregnancy. AB - Fractalkine is a new CX(3)C chemokine that has chemoattractant activity for T cells, monocytes and natural killer (NK) cells. Western blot analysis revealed that fractalkine protein was detected as a 95 kDa band in both the amniotic fluid and the amnion during the second and third trimesters. Immunohistochemistry using an anti-fractalkine polyclonal antibody revealed positive staining of epithelial cells in amnion and trophoblasts in both the second and third trimesters. Neonatal urine also contained detectable amounts of fractalkine. RT-PCR detected fractalkine mRNA transcripts in the amnion. To determine whether fractalkine receptor (CX(3)CR1)-positive cells were present in amniotic fluid and amnion, we performed RT-PCR using specific primers for CX(3)CR1. CX(3)CR1-positive cells had migrated into the amniotic fluid and the amnion. The present findings suggest that fractalkine found in amniotic fluid may contribute to the immunodefence mechanism during pregnancy. PMID- 12569180 TI - Interleukin-1beta stimulates placental leucine aminopeptidase/oxytocinase expression in BeWo choriocarcinoma cells. AB - In addition to prostaglandins, inflammatory cytokines induce uterine contraction via oxytocin (OT). Placental leucine aminopeptidase (P-LAP), an oxytocinase that is identical to cystine aminopeptidase, destroys OT activity. Patients with spontaneous preterm delivery have higher concentrations of inflammatory cytokines and lower P-LAP activities than those with normal delivery. In addition, the P LAP promoter region contains putative binding sites for cytokine-induced transcription factors. We therefore postulated that inflammatory cytokines suppress P-LAP expression and examined this notion using BeWo choriocarcinoma cells cultured in the presence of cytokines. However, interleukin-1beta (IL 1beta) increased P-LAP activity in a time- and dose-dependent manner. Furthermore, Western blot analysis showed a dose-dependent increase of P-LAP proteins. We also detected IL-1 type I receptor mRNA in BeWo cells by RT-PCR. Semi-quantitative RT-PCR and Southern blot analysis showed that IL-1beta also increased P-LAP mRNA, which was abrogated by prior exposure to cycloheximide. Luciferase assays did not reveal any regulatory regions that could explain IL 1beta-induced P-LAP mRNA accumulation within 1.1 kb upstream of the P-LAP gene. Immunohistochemical analysis of human placenta with chorioamnionitis demonstrated prominent P-LAP staining at sites of abundant inflammatory cell infiltration. These findings indicated that prolonged exposure to IL-1beta induces P-LAP in the trophoblasts, possibly via other de-novo protein synthesis, which contradicted our initial hypothesis. PMID- 12569181 TI - First preimplantation genetic diagnosis of hereditary retinoblastoma using informative microsatellite markers. AB - Retinoblastoma is a malignant intra-ocular tumour of developing retina initiated by inactivation of both alleles of the retinoblastoma susceptibility (RB1) gene. This paper reports the first clinical experience of preimplantation genetic diagnosis (PGD) for hereditary retinoblastoma using two highly polymorphic microsatellite markers RB1.20 and D13S284, located within and close to the RB1 gene respectively. Duplex PCRs were tested on more than 300 single lymphocytes from heterozygous individuals at both loci, in order to test the accuracy and reliability of the single-cell protocol. This procedure requires a nested PCR and the analysis of fluorescently labelled PCR products on an automatic DNA sequencer. Amplification efficiency and allele drop-out rates ranged from 96.7 to 98.4%, and 3.7 to 5.4% respectively. This test was found to be accurate and reliable enough to be applied to the study of human blastomeres. Subsequently, this approach was used in a PGD treatment cycle for a couple who already had a child affected with hereditary retinoblastoma and found to be informative for both microsatellite markers. PMID- 12569183 TI - Institute of Medical Ethics prize for the most innovative web publication. PMID- 12569182 TI - Dissociation of steroid receptor coactivator 1 and nuclear receptor corepressor recruitment to the human glucocorticoid receptor by modification of the ligand receptor interface: the role of tyrosine 735. AB - Within the human glucocorticoid receptor (GR) steroid binding pocket, tyrosine 735 makes hydrophobic contact with the steroid D ring. Substitution of tyrosine735 selectively impairs glucocorticoid transactivation but not transrepression. We now show, using both mammalian two-hybrid and glutathione-S transferase pull downs, that such substitutions reduce interaction with steroid receptor coactivator 1, both basally and in response to agonist binding. Using a yeast two-hybrid screen we identified one of the three nuclear receptor interacting domains (NCoR-N1) of nuclear receptor corepressor (NCoR) as interacting with the GR C terminus in an RU486-specific manner. This was confirmed in mammalian two-hybrid experiments, and so we used the NCoR-N1 peptide to probe the GR C-terminal conformation. Substitution of Tyr735phe, Tyr735val, and Tyr735 ser, which impaired steroid receptor coactivator 1 (SRC1) interaction, enhanced NCoR-N1 recruitment, basally and after RU486. RU486 did not direct SRC1 recruitment to any of the GR constructs, and dexamethasone did not allow NCoR-N1 recruitment. Using a glutathione-S-transferase pull-down approach, the NCoR-N1 peptide was found to bind the full-length GR constitutively, and no further induction was seen with RU486, but it was reduced by dexamethasone. As both SRC1 and NCoR are predicted to recognize a common hydrophobic cleft in the GR, it seems that changes favorable to one interaction are detrimental to the other, thus identifying a molecular switch. PMID- 12569184 TI - Consent and confidentiality--where are the limits? An introduction. PMID- 12569185 TI - Some limits of informed consent. AB - Many accounts of informed consent in medical ethics claim that it is valuable because it supports individual autonomy. Unfortunately there are many distinct conceptions of individual autonomy, and their ethical importance varies. A better reason for taking informed consent seriously is that it provides assurance that patients and others are neither deceived nor coerced. Present debates about the relative importance of generic and specific consent (particularly in the use of human tissues for research and in secondary studies) do not address this issue squarely. Consent is a propositional attitude, so intransitive: complete, wholly specific consent is an illusion. Since the point of consent procedures is to limit deception and coercion, they should be designed to give patients and others control over the amount of information they receive and opportunity to rescind consent already given. PMID- 12569186 TI - Confidentiality and its limits: some contributions from Christianity. AB - The issue is whether Christianity, of its nature, would seek to prevent a justifiable breach of confidentiality or could endorse it, under certain circumstances, as the act which is fundamentally more loving or more truthful. The individualistic nature of Western Christianity is noted. The Lutheran theologian Dietrich Bonhoeffer is used to show Christian support for dynamic rather than literal truth telling, and for awareness of the contexts and power relations within which persons stand. PMID- 12569188 TI - Consent and confidentiality in genetics: whose information is it anyway? AB - Against a background of increasing regulation regarding access to medical information and the presentation of patients' confidentiality, the case of genetic information raises interesting questions about whether the application of general rules is appropriate in all situations. Whilst all genetic information is not equally sensitive, some of it is highly predictive. It also allows deductions to be made about other family members. It may not be regarded as particularly sensitive when compared to other types of medical information and those to whom it applies may not be as anxious about preserving their confidentiality as compared with-for example, the prospect of seeing research into cause and cures for rare diseases put in hand. These distinctions also find resonance with the general public. Resolving conflicting tensions will require subtlety, not a blunt "one size fits all" model. PMID- 12569187 TI - Consent and end of life decisions. AB - This paper discusses the role of consent in decision making generally and its role in end of life decisions in particular. It outlines a conception of autonomy which explains and justifies the role of consent in decision making and criticises some misapplications of the idea of consent, particular the role of fictitious or "proxy" consents. Where the inevitable outcome of a decision must be that a human individual will die and where that individual is a person who can consent, then that decision is ethical if and only if the individual consents. In very rare and extreme cases such a decision will be ethical in the absence of consent where it would be massively cruel not to end life in order to prevent suffering which is in no other way preventable. Where, however, the human individual is not a person, as is the case with abortion, the death of infants like Mary (one of the conjoined twins in a case discussed in the paper), or in the very rare and extreme cases of those who have ceased to be persons like Tony Bland, such decisions are governed by the ethics of ending the lives of non persons. PMID- 12569189 TI - Research on the mentally incompetent. AB - The specific problems of consent for the mentally incompetent are reviewed. Scientific research is essential to test the validity of present treatments and to develop new ones. The respective roles of the physician and the researcher have to be clearly defined. The vulnerability of psychiatric patients has to be taken into consideration in such a way that some research can be conducted. It is emphasised that the ethical restrictions for research, although highly justified and necessary, are in part responsible for the relatively slow progress in the application of modern neurosciences to psychiatric diseases. PMID- 12569190 TI - Misled and confused? Telling the public about MMR vaccine safety. Measles, mumps, and rubella. AB - The extraordinary events surrounding the measles, mumps, and rubella (MMR) vaccine in the United Kingdom have not only placed in jeopardy the use of this triple vaccine but have also spread concern to other parts of the world. Examination of the public's worry about MMR vaccine reveals they have been exposed to a range of conflicting views resulting in the feeling of having been misled about the safety of the vaccine. There are various groups and individuals who have legitimate roles in informing the public about such subjects. But is each one behaving in an ethically responsible way? And if confidence falters, vaccine coverage dips, and an outbreak of measles, mumps, or rubella ensues, who, if anyone, will stand and say "I misled them, I confused them, this is my responsibility"? We examine the ethical issues of each group with a voice in the debate about vaccine safety. PMID- 12569191 TI - Common sense and common consent in communicable disease surveillance. AB - The need to protect the public against the spread of communicable (infectious) disease provides a good example of the need for a commonsense approach to the use of confidential data. Laboratories need to notify different professionals in order to trace the sources of outbreaks of infection and eradicate the cause. It is often not possible to obtain consent from individual patients, given the rapid time scale required. In doing so, however, laboratory staff and others would contravene the Data Protection Act in passing on information without consent. Section 60 of the Health and Social Care Act was designed to overcome barriers to research relying on data accumulated in the past and this type of public health work. But this is proving a sluggish procedure. It is an awkward solution to the problem of data use without specific consent. This problem will be overcome only when the public can have sufficient trust in safeguards that are in place to protect their safety and confidentiality so that these important activities can be pursued without specific informed consent. PMID- 12569192 TI - Organ retention and return: problems of consent. AB - This paper explores difficulties around consent in the context of organ retention and return. It addresses the proposals of the Independent Review Group in Scotland on the Retention of Organs at Post Mortem to speak of authorisation rather than consent. Practical problems about whose consent determines disputes in relation to organ retention are explored. If a young child dies and his mother refuses consent but his father agrees what should ensue? Should the expressed wishes of a deceased adult override the objections of surviving relatives? The paper suggests much broader understanding of the issues embedded in organ retention is needed to provide solutions which truly meet families' and society's needs. PMID- 12569193 TI - The requirements of the Data Protection Act 1998 for the processing of medical data. AB - The Data Protection Act 1998 presents a number of significant challenges to data controllers in the health sector. To assist data controllers in understanding their obligations under the act, the Information Commissioner has published guidance, The Use and Disclosure of Health Data, which is reproduced here. The guidance deals, among other things, with the steps that must be taken to obtain patient data fairly, the implied requirements of the act to use anonymised or psuedonymised data where possible, an exemption applicable principally to records based research, the right of patients to object to the processing of their data, and the interface of the act and the common law duty of confidence. PMID- 12569194 TI - Confidentiality and the duties of care. AB - Doctors have an ethical and legal duty to respect patient confidentiality. We consider the basis for this duty, looking particularly at the meaning and value of autonomy in health care. Enabling patients to decide how information about them is disclosed is an important element in autonomy and helps patients engage as active partners in their care. Good quality data is, however, essential for research, education, public health monitoring, and for many other activities essential to provision of health care. We discuss whether it is necessary to choose between individual rights and the wider public interest and conclude that this should only rarely be necessary. The paper makes some recommendations on practical steps which could help ensure that good quality information is available for work which benefits society and the public health, while still enabling patients' autonomy to be respected. PMID- 12569195 TI - Incapacity to give informed consent owing to mental disorder. AB - What renders some mentally disordered patients incapable of informed consent to medical interventions? It is argued that a patient is incapable of giving informed consent owing to mental disorder, if a mental disorder prevents a patient from understanding what s/he consents to; if a mental disorder prevents a patient from choosing decisively; if a mental disorder prevents a patient from communicating his/her consent; or if a mental disorder prevents a patient from accepting the need for a medical intervention. This paper holds that a patient's capacity to give informed consent should be assessed clinically by using these conditions necessary for informed consent, and should be assessed specifically for each intervention and specifically at the time when the consent has to be given. The paper considers patients' incapacity to give informed consent to treatment, to give informed consent to be examined clinically, and to give informed consent to participate in research. PMID- 12569196 TI - Knowledge of the legislation governing proxy consent to treatment and research. AB - OBJECTIVE: To assess the knowledge of four groups of individuals regarding who is legally authorised to consent to health care or research involving older patients. DESIGN: A provincewide postal survey. SETTING: Province of Quebec, Canada. PARTICIPANTS: Three hundred older adults, 434 informal caregivers of cognitively impaired individuals, 98 researchers in aging and 136 members of research ethics boards (REBs). MEASUREMENTS: Knowledge was assessed through a pretested postal questionnaire comprising five vignettes that describe hypothetical situations involving an older adult who requires medical care or is solicited for research. The respondent had to identify the person who is legally authorised to provide consent. RESULTS: Nearly 80% of all respondents provided the correct answer when the hypothetical scenario depicted a person who was competent to consent or incompetent but legally represented. Knowledge was worse (from 2% among older adults to 44% among REB members) for the scenario describing a research situation that involved an incompetent adult without a legal guardian. CONCLUSION: The observed lack of knowledge raises doubts about the ability of current legislation to truly protect the rights of older adults with diminished decision making capacity. It points to the need for educational programmes aimed at increasing public awareness of the legislation put in place for those requiring special protection. PMID- 12569197 TI - Relatives' attitudes towards informing patients about the diagnosis of Alzheimer's disease. AB - OBJECTIVES: To evaluate relatives' attitudes towards informing patients with Alzheimer's disease (AD) about their diagnosis. SETTING: A university hospital in Italy. METHODS: The closest relatives of each of 71 subjects diagnosed for the first time as having AD were interviewed, using a semistructured questionnaire. Spontaneous requests by relatives not to communicate issues concerning the diagnosis were also recorded. RESULTS: Forty three (60.6%) relatives spontaneously requested that patients not be fully informed. After being interviewed, nobody thought that the patient should be given all the information. Justifications were related to the fear of the onset or worsening of depressive symptoms in the patient. CONCLUSIONS: In Italy relatives' opposition to informing AD patients appears to be common. Knowledge of the relatives' attitudes may be useful for clinicians but disclosure of diagnosis should be based on the clinical evaluation of the patient and on a prudent evaluation of the relationship between the patient and her/his relative caregiver. PMID- 12569198 TI - Psychiatric research: what ethical concerns do LRECs encounter? A postal survey. Local research ethics committees. AB - BACKGROUND AND METHODS: Psychiatric research can occasionally present particular ethical dilemmas, but it is not clear what kind of problems local research ethics committees (LRECs) actually experience in this field. We aimed to assess the type of problems that committees encounter with psychiatric research, using a postal survey of 211 LRECs. RESULTS: One hundred and seven (51%) of those written to replied within the time limit. Twenty eight (26%) experienced few problems with psychiatric applications. Twenty six (24%) emphasised the value of a psychiatric expert on the committee. The most common issues raised were informed consent (n=64, 60%) and confidentiality (n=17, 16%). The use of placebos (and washout periods) (n=18, 17%), the validity of psychiatric questionnaires (n=16, 15%) and overuse of psychiatric "jargon" (n=14, 13%) in psychiatric applications also raised concern. CONCLUSIONS: Our results suggest that LRECs have specific concerns regarding methodology, consent, and confidentiality in psychiatric research, and that they find psychiatric input invaluable. PMID- 12569200 TI - American biofutures: ideology and utopia in the Fukuyama/Stock debate. AB - Francis Fukuyama, in his Our Posthuman Future, and Gregory Stock, in his Redesigning Humans, present competing versions of the biomedical future of human beings, and debate the merits of more or less stringent regimes of regulation for biomedical innovation. In this article, these positions are shown to depend on a shared discourse of market liberalism, which limits both the range of ends for such innovation discussed by the authors, and the scope of their policy analyses and proposals. A proper evaluation of the human significance and policy imperatives for biomedical innovation needs to be both more utopian in its imagination, and more sophisticated in its political economy. In essence, the Fukuyama/Stock debate tells us more about current US political ideology than it does about the morality of human genetic and biopsychological engineering. PMID- 12569202 TI - Exercise training in women with heart disease: influence of hormone replacement therapy. AB - PURPOSE: To examine the central and peripheral cardiovascular effects of exercise training in postmenopausal women with CAD with and without hormone replacement therapy (HRT and N-HRT). METHODS: Thirty-eight female cardiac patients referred for cardiac rehabilitation were divided into HRT ( N= 18) or N-HRT (N = 20) groups. Peak oxygen uptake (VO2) peak and ventilatory anaerobic thresholds (AT) were determined, in addition to submaximal cardiac output (Q). Peripheral measures of resting and peak ischemic blood flows (BF) were also measured. Measurements were all repeated after 12 and 26 wk of exercise training consisting of 1 h of walking at 75-80% of the measured VO2peak at baseline (T1) for 5 d.wk( 1). RESULTS: VO2peak mL.kg(-1).min(-1) at baseline (14.9 +/- 0.4) increased by 5% after 12 wk (15.6 +/- 0.4) and significantly by 15% (17.2 +/- 0.5) after 26 wk of exercise training (P < 0.001). The HRT group was significantly younger than the N HRT group (58 vs 65 yr; P < 0.01) and had significantly higher VO2peaks at baseline (15.7 vs 14.2 mL.kg(-1).min(-1); P < 0.05), yet either did not influence changes in other variables. At fixed submaximal work rates, there was a significant training bradycardia ( P < 0.01), but insignificant changes in Q or stroke volume regardless of HRT status. Resting and peak ischemic calf BF and vascular conductance increased significantly ( P < 0.001) at 12 and 26 wk, with no difference found according to HRT status. CONCLUSIONS: The cardiovascular responses to training in postmenopausal women with CAD appear to be consistent regardless of HRT status and dominated by peripheral adaptations. PMID- 12569201 TI - Identification of an endogenous ligand that activates pregnane X receptor mediated sterol clearance. AB - The nuclear receptor PXR (pregnane X receptor) is a broad-specificity sensor that recognizes a wide variety of synthetic drugs and xenobiotic agents. On activation by these compounds, PXR coordinately induces a network of transporters, cytochrome P450 enzymes, and other genes that effectively clear xenobiotics from the liver and intestine. Like PXR, the majority of its target genes also possess a broad specificity for exogenous compounds. Thus, PXR is both a sensor and effector in a well integrated and generalized pathway for chemical immunity. Although it is clear that PXR responds to numerous foreign compounds, it is unclear whether it possesses an endogenous ligand. To address this issue, we noted that there is substantial overlap in the substrate specificities of PXR and its critical CYP3A target gene. This prompted us to ask whether endogenous CYP3A substrates also serve as PXR ligands. We demonstrate that 5beta-cholestane 3alpha,7alpha,12alpha-triol (triol), a cholesterol-derived CYP3A substrate, is a potent PXR agonist that effectively induces cyp3a expression in mice. This defines a critical salvage pathway that can be autoinduced to minimize triol accumulation. In contrast, triol can accumulate to very high levels in humans, and unlike mice, these people develop the severe clinical manifestations of cerebrotendinous xanthomatosis. The reason for these dramatic species differences has remained unclear. We now demonstrate that triol fails to activate human PXR or induce the CYP3A-salvage pathway. This explains why humans are more susceptible to sterol accumulation and suggests that synthetic ligands for human PXR could be used to treat cerebrotendinous xanthomatosis and other disorders of cholesterol excess. PMID- 12569204 TI - Muscle pain in athletes with locomotor disability. AB - PURPOSE: Athletes with locomotor disabilities (LDA) participate in many competitive sport activities, yet little is known about sport-related muscle pain (SRMP). This study assessed the prevalence, determinants, and main characteristics of SRMP in LDA. METHODS: A cross-disability epidemiological survey was used to investigate the occurrence of SRMP during the previous year by using a questionnaire administrated by medical doctors. SRMP was defined as any muscle pain experienced during the past 12 months that either occurred during sport activity (training or competition) and/or was reported as a consequence of physical exercise, causing discomfort for at least 1 d and not related to systemic disease. A total of 227 LDA were recruited randomly from the population (567 LDA) who participated in selected National sports events (including swimming, athletics, wheelchair basketball matches, and others) organized by the Italian Federation of Sports for Disabled. Collected data were statistically analyzed with univariate and multivariate logistic regression models to identify possible determinants of SRMP, through the estimate of the prevalence odds ratios. RESULTS: The SRMP period prevalence rate was equal to 50.7% (95% confidence interval (44.0-57.4%)), ranging from 47.0% (swimmers) to 58.8% (basketball athletes). In 71.1% of cases, SRMP lasted less than 1 wk and only 8.7% experienced pain for more than 1 month. SRMP had a higher prevalence in amputees (75.0%) and spinal cord injured LDA (58.1%) than the other groups. There was increased prevalence rate of SRMP with increased training volume. The multivariate logistic regression model showed disorder type, body mass index, and training volume as determinants of SRMP. CONCLUSIONS: Prospective studies could be devised to assess the role of anthropometric characteristics and training volume as risk factors of SRMP. PMID- 12569203 TI - A single bout of walking exercise enhances endogenous fibrinolysis in stroke patients. AB - PURPOSE: Impaired endogenous fibrinolysis is an important predictor for increased risk of stroke and myocardial infarction. Acute exercise can enhance fibrinolysis, primarily by stimulating short-term increases in plasma tissue plasminogen activator (tPA), which is postulated to protect against atherothrombotic events. No prior studies have examined the fibrinolytic response to exercise in stroke survivors despite their high risk for recurrent stroke and myocardial infarction. The purpose of this study was to assess the fibrinolytic response to acute submaximal exercise in chronic hemiparetic stroke patients. METHODS: Eighteen (16 men, 2 women) untrained stroke patients with chronic hemiparetic gait deficits volunteered for participation in this single session exercise study. Fasting blood samples for determination of tPA and plasminogen activator inhibitor (PAI-1) enzyme activities were obtained before, immediately after, and 60 min after submaximal treadmill walking. Patients walked at 60% maximal heart rate reserve (low-moderate intensity) for a cumulative total of 20 min. RESULTS: The exercise bout increased tPA activity by 79% (P < 0.01) and decreased PAI-1 activity by 18% (P < 0.01). At 1 h after completing the walking exercise, plasma tPA activity levels were still significantly elevated (43%,P < 0.01), and PAI-1 activity levels were 25% lower (P < 0.01) than baseline. CONCLUSIONS: These findings demonstrate that a single bout of aerobic walking exercise can improve fibrinolysis profiles in chronic stroke patients. Significant increases in endogenous tPA and reductions in PAI-1 activity persist for at least 1 h after exercise cessation. The implications are that alterations in physical activity during the day may modify clot lysing potential, thereby affecting atherothrombotic risk. PMID- 12569205 TI - Exercise-induced weight loss preferentially reduces abdominal fat. AB - PURPOSE: To investigate whether abdominal fat is reduced in response to substantial weight loss induced by exercise in young obese men. METHODS: Thirty obese men (mean age 19.8 +/- 0.6 yr) were evaluated before (pretraining) and after (posttraining) 4 months of regimented training in the Singapore Armed Forces. There were 30 obese male subjects (mean age 19.2 +/- 1.3 yr) without training who were monitored as control subjects. Fat free mass (FFM), fat mass, and percent body fat were determined from skinfold measurements. Differences between pre- and posttraining responses were analyzed with a paired -test. RESULTS: Subjects lost 12.0 +/- 3.6 kg (P < 0.001) from pre- to posttraining, which was attributable to a reduction in fat mass (P < 0.001), as FFM was unchanged. Both waist circumference (WC) and hip circumference (HC) decreased (P < 0.01), the reduction in WC (13.7%) being greater than the reduction in HC (7.7%) as reflected by the decrease in waist-to-hip ratio (WHR; P < 0.001). These data reveal that large exercise-induced weight losses are associated with maintenance of FFM. The significant reduction in WHR indicates a greater mobilization of abdominal fat and a preferential loss of fat from this region. CONCLUSIONS: Large exercise-induced weight loss is associated with a preferential reduction in abdominal fat and a corresponding maintenance of FFM. Such an effect on body composition should reduce disease risk and the eventual weight regain that typically follows diet-induced weight losses with obese subjects. PMID- 12569206 TI - The relationship of left ventricular to femoral artery structure in male athletes. AB - PURPOSE: In humans, cardiac morphological adaptations to athletic training have been exhaustively described, while the existence of morphologic (adaptive) changes in the peripheral vessels are less well known. Therefore, the scope of the present study was 1) to assess the existence of morphologic changes in the peripheral vessels, such as the common femoral artery (CFA), in athletes engaged in different types of sports; and 2) to assess the relationship existing between the cardiac and CFA changes in highly endurance- ( N = 16) and strength-trained athletes ( N = 15), and in sedentary controls ( N = 20). METHODS: The cross sectional diameters of the left ventricular (LV) posterior wall and cavity and CFA diameter and intima-media thickness were determined by M-mode echocardiography and ultrasound, respectively. RESULTS: CFA intima-media thickness in the endurance-trained (0.33 +/- 0.03 mm.m(-2) ) was similar to the strength-trained athletes (0.34 +/- 0.05 mm.m(-2) ) and both greater (P < 0.01) compared with the controls (0.27 +/- 0.03 mm.m(-2) ). CFA diameter in endurance trained athletes (5.6 +/- 0.6 mm.m(-2) ) was highest among groups (strength trained athletes 4.2 +/- 0.4 mm.m(-2), controls 4.2 +/- 0.5 mm.m(-2) ). Thus, the relative CFA wall thickness (2.intima-media thickness/CFA diameter) was significantly higher (P < 0.01) in the strength-trained (0.16 +/- 0.02) than in the endurance-trained athletes (0.13 +/- 0.02) and the controls (0.13 +/- 0.02), respectively. The relative LV posterior wall thickness (2.LV posterior wall thickness/LV diameter) was not significantly different among the groups. LV posterior wall thickness was significantly related to CFA intima-media thickness (r = 0.31, P < 0.05) and LV diameter to CFA diameter (r = 0.55, P < 0.01). CONCLUSIONS: The study shows a physiological relationship between the LV and CFA wall and cavity dimensions in endurance-trained athletes and controls, respectively. The greater relative wall thickness of the strength-trained athletes may be related to different influencing factors, e.g., increased blood pressure. PMID- 12569208 TI - Muscle metaboreflex-induced increases in stroke volume. AB - PURPOSE: Accumulation of by-products of metabolism within skeletal muscle may stimulate sensory nerves, thus evoking a pressor response named muscle metaboreflex. The aim of this study was to evaluate changes in central hemodynamics occurring during the metaboreflex activation. METHODS: In seven healthy subjects, the metaboreflex was studied by postexercise regional circulatory occlusion at the start of the recovery from a mild rhythmic forearm exercise. Central hemodynamics was evaluated by means of impedance cardiography. RESULTS: The main findings of this study were that, with respect to rest, the metaboreflex: 1) raised mean blood pressure (+13%; P < 0.01); 2) enhanced myocardial contractility (-12% in preejection period/left ventricular ejection time ratio; P < 0.01); 3) prolonged diastolic time (+11%; P < 0.01); 4) increased stroke volume (+ 10%; P < 0.05); and 5) increased cardiac output (+6%; P < 0.05). These responses were present neither during recovery without circulatory occlusion nor during circulatory occlusion without prior exercise. Moreover, the metaboreflex did not affect systemic vascular resistance and induced bradycardia with respect to recovery without circulatory occlusion. CONCLUSION: These results suggest that the blood pressure response during metaboreflex activation after mild rhythmic exercise is strongly dependent on the capacity to increase cardiac output rather than due to increased vascular resistance. PMID- 12569210 TI - Muscle metabolic function, exercise performance, and weight gain. AB - PURPOSE: The study purpose was to determine the relationship: 1) of muscle metabolism to exercise performance and 2) of exercise performance to rate of weight gain. METHODS: Eighty-three black and white premenopausal women were evaluated for maximum oxygen uptake (VO2max ), isometric quadriceps and triceps surae strength, and 31P magnetic resonance spectroscopy of calf muscle metabolic capacity. Rate of weight gain was determined 1 yr later. Multiple regression was used to model dependent variables. RESULTS: Muscle aerobic capacity and strength of the quadriceps muscle independently contributed to endurance time on the treadmill (ET) in one model (overall R = 0.47, P < 0.01), and VO2max and strength of the quadriceps muscle independently contributed to ET in another model (R = 0.85, P < 0.001). In models of muscle strength, maximum creatine kinase activity and maximum anaerobic glycolytic rate independently contributed to triceps surae strength, after adjusting for triceps surae cross-section area (R = 0.63, P < 0.001). In another model, maximum creatine kinase activity was related to quadriceps strength independent of leg lean tissue (R = 0.31, P < 0.05). Rate of weight gain was related to muscle metabolic economy (r = -0.25, = 0.04), quadriceps strength (r = -0.34, P < 0.01), VO2max (r = -0.22, = 0.04), and ET (r = -0.21, = 0.04). Rate of weight gain was modeled by muscle metabolic economy, VO2max, and quadriceps strength (R = 0.48, P < 0.01). CONCLUSIONS: Implications of findings are 1) greater strength and aerobic fitness-at the muscle and whole body levels-improve endurance; 2) greater muscle anaerobic metabolism is associated with greater muscle strength, independent of muscle size; and 3) greater exercise endurance reduces weight gain. PMID- 12569211 TI - Specificity and reversibility of inspiratory muscle training. AB - PURPOSE: The purpose of this study was to evaluate the pressure-flow specificity of adaptations to inspiratory muscle training (IMT), in addition to the temporal effects of detraining and reduced frequency of training upon these adaptations. METHODS: Twenty-four healthy subjects were assigned randomly to one of four groups (A: low-flow-high-pressure IMT; B: high-flow-low-pressure IMT; C: intermediate flow-pressure IMT; and D: no IMT). Subjects performed IMT 6 d.wk(-1) for 9 wk, and inspiratory muscle function was evaluated at baseline and every 3 wk. Groups A, B, and C were then assigned randomly to either a maintenance group (M) (IMT 2 d.wk(-1) ) or a detraining group (DT) (no IMT). Inspiratory muscle function was reassessed at 9 and 18 wk post-IMT. RESULTS: At 9 wk, group A exhibited the largest increase in pressure, B a large increase in flow, C more uniform increases in pressure and flow, and D no changes in pressure or flow. Maximum inspiratory muscle power increased in groups A, B, and C by 48 +/- 3%, 25 +/- 3%, and 64 +/- 3%, respectively (mean +/- SEM, P < or = 0.01). Maximum rate of pressure development increased in groups A, B, and C by 59 +/- 1%, 10 +/- 1%, and 29 +/- 1%, respectively ( P < or = 0.01). A decrease in inspiratory muscle function was observed at 9 wk post-IMT in DT. Inspiratory muscle function plateaued between 9 and 18 wk but remained above pre-IMT values. Group M retained the improvements in inspiratory muscle function. CONCLUSION: These data support the notion of pressure-flow specificity of IMT. Detraining resulted in small but significant reductions in inspiratory muscle function. Reducing training frequency by two thirds allowed for the maintenance of inspiratory muscle function up to 18 wk post-IMT. PMID- 12569212 TI - Eccentric muscle strength in functional ankle instability. AB - PURPOSE: To investigate muscle strength in subjects with unilateral functional ankle instability, with the specific aim of determining whether eccentric strength deficits exist. METHODS: Isokinetic invertor and evertor torques were assessed eccentrically and concentrically at speeds of 60 and 120 degrees.s(-1) in 16 subjects with unilateral ankle instability. Absolute peak torque values were compared between the injured and noninjured limbs to determine whether strength deficits existed. RESULTS: No eccentric or concentric evertor strength deficit was found in the injured limb. Interestingly, for the injured limb, a significant interaction effect was found whereby eccentric inversion strength was reduced. CONCLUSION: Functional ankle instability is not associated with deficits in evertor strength. Deficits in eccentric invertor strength were found, and this may contribute to the symptoms of functional ankle instability. Weak invertors may contribute to functional ankle instability because of a reduced ability to assist in controlling lateral displacement of the shank over the weight-bearing foot. Excess lateral postural sway outside of the base of support results in the foot's medial border rising from the floor once the limits of closed chain eversion is reached, which may result in the foot being forced into rapid inversion. PMID- 12569213 TI - Isometric training lowers resting blood pressure and modulates autonomic control. AB - PURPOSE: This study examined the effects of isometric handgrip training on resting arterial blood pressure, heart rate variability, and blood pressure variability in older adults with hypertension. METHODS: Nine subjects performed four 2-min isometric handgrip contractions at 30% of the maximum voluntary contraction force, 3 d.wk(-1) for 10 wk, and eight subjects served as controls. RESULTS: After training, there was a significant reduction in resting systolic pressure and mean arterial pressure. In addition, power spectral analysis of heart rate variability demonstrated that the low frequency: high frequency area ratio tended to decrease. CONCLUSIONS: It is concluded that isometric training at a moderate intensity elicits a hypotensive response and a simultaneous increase in vagal modulation in older adults with hypertension. PMID- 12569214 TI - Endurance training reduces end-exercise VO2 and muscle use during submaximal cycling. AB - INTRODUCTION: End-exercise VO2 during heavy, constant-load exercise is reduced after endurance training, due to an attenuated VO2 slow component. PURPOSE/METHODS: To determine whether the training-induced reduction in end exercise VO2 was associated with reduced muscle use, we measured VO2 and T2 changes in magnetic resonance images in the final minute of two 15-min constant load cycle rides, one above lactate threshold and the other below lactate threshold. These measures were repeated after a 4-wk period in eight subjects who trained on a cycle ergometer and seven controls. RESULTS: There were no changes in end-exercise VO2 or active muscle after training in either group during low intensity cycling, in which no VO2 slow component was present. During high intensity cycling, in which there was a slow component before training, the training group experienced a significant reduction (P < 0.05) in end-exercise VO2 (2625 +/- 673; 2567 +/- 605 mL.min (-1) and the T2 of the vastus lateralis (35.6 +/- 1.4; 34.5 +/- 0.9 ms). CONCLUSION: These results support the hypothesis that reduction in end-exercise VO2 (and the VO2 slow component) after training is due to reduced muscle use during heavy, constant load cycling. PMID- 12569215 TI - Evaluation of stress responses to interval training at low and moderate altitudes. AB - PURPOSE: The purpose of the present field study was to explore whether extensive interval training (IT) performed with a similar behavior of blood lactate (LA) at an altitude of 1800 m (ALT) and near sea level (SL) goes along with a comparable hormonal, metabolic, and acute phase response in highly trained endurance athletes. METHODS: Twelve distance runners (VO2 64.6 +/- 6.9 mL.kg(-1) ) performed IT (10 x 1000 m, 2-min rest) at SL with a running velocity (V) corresponding to 112% of the individual anaerobic threshold (IAT). After an acclimatization period of 7 d, IT was repeated with a lower V (107% IAT) at ALT. Blood samples were drawn at rest, 0, 0.3, 3, and 24 h after IT. LA during IT was similar at SL and ALT (5.4 +/- 1.3/5.3 +/- 1.2 mmol.L(-1)), whereas HR tended to be higher at SL. RESULTS: Postexercise rises in plasma noradrenaline (NA), NA sulfate, adrenaline, glucose, interleukin-6 (IL-6), and neutrophils were significantly more pronounced at ALT. The increase of cortisol and human growth hormone showed an insignificant trend toward higher values at ALT. A slight but significant increase of plasma erythropoietin was only apparent after IT at ALT. No differences between either condition were observed for exercise-related changes in free fatty acids, IL-8, lympho-, or monocyte counts. CONCLUSIONS: In spite of a matched accumulation pattern of LA between ALT and N, stress responses, such as sympathetic activation and hepatic glucose release, still appear to be greater at ALT. This additional impact of moderate ALT on the stress response to IT should be taken into account if repeated training sessions are performed within a short period of time. PMID- 12569216 TI - The effect of cardiorespiratory fitness and obesity on cancer mortality in women and men. AB - PURPOSE: The purpose of this study was to determine the independent and combined effects of cardiorespiratory fitness and obesity on all-cause cancer mortality for women and men. METHODS: Using the Lipids Research Clinics Prevalence Study, we examined the relationship of fitness and obesity on cancer mortality among 2585 women and 2890 men followed from 1972-1976 to 1998. Cardiorespiratory fitness was measured using a treadmill test and obesity was assessed using body mass index (BMI) calculated from measured height and weight. Gender-specific hazard ratios (HR) were calculated from proportional hazard models, which included covariates for age, education, smoking, alcohol intake, Keys score, and menopause (women only). RESULTS: Adjusted cancer mortality was significantly lower in the most fit quintile relative to the other four quintiles for men (HR = 0.47; 95% CI, 0.27-0.81) but not for women (HR = 0.84; 95% CI, 0.52-1.36). Adjusted cancer mortality was significantly higher in the highest BMI quintile relative to the other four BMI quintiles for women (HR = 1.49; 95% CI, 1.06-2.09) but not for men (HR = 1.05; 95% CI, 0.77-1.43). Further adjustment for BMI on fitness and adjustment for fitness on BMI did not meaningfully change the HR. There were no significant interactions between fitness and obesity in predicting cancer mortality for either women or men. CONCLUSION: In this study, high fitness was a stronger predictor of cancer mortality in men, whereas high BMI was a stronger predictor of cancer mortality in women. PMID- 12569217 TI - Lifetime physical activity and breast cancer risk in pre- and postmenopausal women. AB - PURPOSE: This research examined associations between leisure time and occupational physical activity (PA) across the lifespan and pre- and postmenopausal breast cancer. METHODS: In a population-based case-control study, 301 premenopausal cases, 316 premenopausal controls, 439 postmenopausal cases, and 494 postmenopausal controls, 40- to 85-yr-old reported time spent in exercise or sports strenuous enough to sweat and miles walked per week for time periods 2, 10, and 20 yr before the interview and at age 16. Lifetime occupational history was obtained. Jobs were coded according to the National Cancer Institute's PA job matrix. RESULTS: Strenuous PA was generally associated with a reduced breast cancer risk. Among women categorized as active at all four periods [at least 91+ h.yr(-1) (1.75+ h.wk(-1) avg)], a strong, significant protective effect was observed in postmenopausal [odds ratio (OR) 0.50 (0.28-0.90)] but not in premenopausal women [OR 1.06 (0.54-2.08)]. A strong protective effect was observed for activity performed 20 yr prior, in both pre- and postmenopausal women, although CIs overlapped for different time periods. Using women who reported no strenuous activity as the referent, OR (95% CIs) for the highest PA category [182+ h.yr(-1) (3.5 h.wk(-1) avg)] 20 yr ago were 0.57(0.31-1.05) and 0.51(0.31-0.83) for pre- and postmenopausal women, respectively. Walking was generally unrelated to risk. There was some indication of increased risk for the upper category of occupational PA for postmenopausal women, perhaps related to other industrial occupational exposures. CONCLUSION: Our results suggest a modest protective effect of strenuous leisure time PA on breast cancer risk in both pre- and postmenopausal women. The effects appear strongest for those active at least 20 yr prior and among postmenopausal women who were consistently active throughout their lifetime. PMID- 12569218 TI - Daughters and mothers exercising together: effects of home- and community-based programs. AB - PURPOSE: This pilot study compares the effectiveness of home- and community-based physical activity interventions that target mothers and daughters to increase physical activity and improve health-related fitness. METHODS: Mothers (45.18 +/- 7.49 yr) and daughters (15.41 +/- 1.33 yr) were randomly assigned to a community based (CB) (N = 20 participants) or home-based (HB) (N = 14 participants) program. CB participants attended three instructor-led sessions per week for 12 wk. HB participants were asked to participate in 3 sessions per week for 12 wk in a program similar to the CB program. The main difference between the programs was that CB activities were completed at a fitness facility within a university and HB activities were completed in or near the home. Before and after the intervention, changes in health-related fitness and physical activity were assessed. A series of 2 (group assignment) x 2 (time) ANOVAs were conducted to assess changes separately for mothers and daughters. RESULTS: CB participants attended 77% of the sessions, and none of the pairs dropped out. HB participants completed 70% of the recommended sessions, and three pairs dropped out. Mothers and daughters in both groups significantly increased their participation in aerobic, muscular strength, and flexibility activities (P = 0.02 to 0.000). Daughters in both groups significantly improved their muscular endurance (sit ups,P = 0.000). Mothers in both groups improved their muscular strength (push ups, P = 0.003), muscular endurance (sit-ups, P = 0.000), flexibility (sit-and reach, P = 0.008), and aerobic capacity (1-mile walk, P = 0.002). Positive changes in diastolic blood pressure also occurred (P = 0.008). CONCLUSION: Mothers and daughters responded positively to CB and HB physical activity programs. Home-based physical activity programming is a cost-effective means to increase physical activity and improve health-related fitness in these groups. PMID- 12569219 TI - Training and bioenergetic characteristics in elite male and female Kenyan runners. AB - PURPOSE: This study compares the training characteristics and the physical profiles of top-class male and female Kenyan long-distance runners. METHOD: The subjects were 20 elite Kenyan runners: 13 men (10-km performance time: 10-km performance time of 28 min, 36 s +/- 18 s) and 7 women (32 min, 32 s +/- 65 s). The male runners were separated into high-speed training runners (HST: N = 6) and low-speed training runners (LST: N = 7) depending on whether they train at speeds equal or higher than those associated with the maximal oxygen uptake (vVO2max ). All but one woman were high-speed training runners (female HST: N = 6). Subjects performed an incremental test on a 400-m track to determine VO2max, vVO2max, and the velocity at the lactate threshold (vLT). RESULTS: Within each gender among the HST group, 10-km performance time was inversely correlated with vVO2max (rho = -0.86, P = 0.05, and rho = -0.95, P = 0.03, for men and women, respectively). HST male runners had a higher VO2max, a lower (but not significantly) fraction of vVO2max (FVO2max ) at the lactate threshold, and a higher energy cost of running (ECR). Among men, the weekly training distance at vVO2max explained 59% of the variance of vVO2max, and vVO2max explained 52% of the variance of 10-km performance time. Kenyan women had a high VO2max and FVO2max at vLT that was lower than their male HST counterparts. ECR was not significantly different between genders. CONCLUSION: The velocity at the VO2max is the main factor predicting the variance of the 10-km performance both in men and women, and high intensity training contributes to this higher VO2max among men. PMID- 12569221 TI - Individual effects of stride length and frequency on shock attenuation during running. AB - Shock attenuation during running is the process of absorbing impact energy due to the foot-ground collision, reducing shock wave amplitude between the foot and head. Shock attenuation is affected by changes in stride length and stride frequency, but it is not clear whether either parameter individually affects shock attenuation. PURPOSE: To identify the independent affects of stride length (SL) and stride frequency (SF) on shock attenuation. METHODS: Subjects ( N = 10) completed three experiments consisting of SL and SF manipulations relative to preferred stride length (PSL) and frequency (PSF). During experiment 1, stride length was manipulated (+15% PSL, PSL, -15% PSL) while stride frequency was always set to PSF. During experiment 2, stride frequency was manipulated (+15% PSF, PSF, -15% PSF) while stride length was always set to PSL. During experiment 3, stride length and stride frequency were manipulated concurrently (+10% PSL/ 10% PSF, PSL/PSF, and -10% PSL/+10% PSF). Running velocity was always the product of stride length and stride frequency. Transfer functions were calculated using tibial and forehead surface mounted accelerometer data to represent shock attenuation. RESULTS: Shock attenuation changed only when stride length changed ( P < 0.05). Specifically, shock attenuation increased as stride length increased. CONCLUSION: It was concluded that changes in stride length not stride frequency affected shock attenuation. PMID- 12569222 TI - Effect of shoe inserts on kinematics, center of pressure, and leg joint moments during running. AB - PURPOSE: The purposes of this project were to assess the effect of four different shoe inserts on the path of the center of pressure (COP), to quantify the effect of these inserts on selected knee joint moments during running, and to assess the potential of COP data to predict the effects of inserts/orthotics on knee joint moments. METHODS: Kinematics for the lower extremities, resultant ankle and knee joint moments, and the path of the COP were collected from the right foot of 15 male subjects while running heel-toe with five different shoe inserts (full or half with 4.5-mm postings). RESULTS: Individual movement changes with respect to the neutral insert condition were typically small and not systematic. Significant changes for the path of the COP were registered only for the full lateral insert condition with an average shift toward the lateral side. The mediolateral shift of the COP was not consistent for the full medial and the two half-shoe inserts. The subject-specific reactions to the inserts' intervention in the corresponding knee joint moments were typically not consistent. Compared with the neutral insert condition, subjects showed increases or decreases of the knee joint moments. The correlation between the individual COP shifts and the resultant knee joint moment was generally small. CONCLUSION: The results of this study showed that subject-specific reactions to the tested inserts were often not as expected. Additionally, reactions were not consistent between the subjects. This result suggests that the prescription of inserts and/or orthotics is a difficult task and that methods must be developed to test and assess these effects. Such methods, however, are not currently available. PMID- 12569223 TI - Effect of monitor placement and of activity setting on the MTI accelerometer output. AB - PURPOSE: To examine the effect of monitor placement (hip vs back) and of activity setting (treadmill vs track) on the output from the Manufacturing Technology Inc. (MTI), activity monitor (model WAM 7164). METHODS: In a laboratory study, 28 subjects (14 men, 14 women) walked at a normal pace, walked at a fast pace, and jogged at a comfortable pace on an indoor track. These activities were repeated on a treadmill using the individual speeds from the track locomotion. Oxygen uptake was measured simultaneously using a portable metabolic system. One activity monitor was worn on the hip and one on the lower back. In a field study, 34 subjects (18 men, 16 women) each wore two monitors (hip and low back placement) for seven consecutive days. In the laboratory study, ANOVA showed significant effects of placement ( P = 0.009) and setting ( P < 0.001), indicating that activity counts differ between different body sites and different settings (track vs treadmill). Gross energy expenditure predictive equations were developed and thereafter evaluated in the field study. Time spent at moderate and vigorous intensity of physical activity was 38% and 85% ( P < 0.001) higher when calculated from the treadmill-based equations as compared to the track-based equations. Free-living physical activity estimates were not affected by the placement. CONCLUSION: The relationship between activity counts and energy expenditure during laboratory locomotion is placement and setting-specific. When habitual physical activity is assessed in free-living subjects, the treadmill derived relationship between energy expenditure and activity counts may overestimate time spent at moderate intensity of physical activity, whereas the placement of the monitor does not influence on the interpretation of the data. PMID- 12569224 TI - Bicycle seat designs and their effect on pelvic angle, trunk angle, and comfort. AB - PURPOSE: To examine whether bicycle seats with anterior-medial cutouts influence pelvic angle, trunk angle, and comfort in female subjects during cycling. METHODS: Twenty female cyclists pedaled a stationary bicycle with their hands on the tops and drops of the handlebars under three different saddle conditions (standard, partial, and complete cutout designs). Pelvic angle was measured using an inclinometer attached to a caliper whereas trunk angle was quantified from digitization of video images. Comfort level was assessed subjectively by having participants rank the saddles from most to least comfortable. RESULTS: Anterior pelvic tilt angles for the partial and complete cutout saddles were 8% and 16% greater, respectively, than values for the standard saddle condition ( P < 0.05). Trunk flexion angles were greater for the complete versus standard and partial cutout designs ( P< 0.05). Participants displayed a 77% greater anterior pelvic tilt angle and an 11% greater trunk flexion angle in the drop versus top handlebar positions ( P < 0.05). A total of 55% of the subjects ranked the partial cutout saddle as the most comfortable, and 30% ranked the standard saddle as the most comfortable. CONCLUSIONS: These data indicate that partial and complete cutout saddle designs may increase anterior pelvic tilt, and saddles with a complete cutout design may increase trunk flexion angles under select cycling conditions. A saddle with a partial cutout design may be more comfortable than a standard or complete cutout saddle design. PMID- 12569225 TI - Concurrent validation of the OMNI perceived exertion scale for resistance exercise. AB - The criterion variables were total weight lifted (Wttot ) determined separately for women and men during BC and KE, and blood lactic acid concentration ([Hla]) determined for a combined female ( N = 10) and male ( N = 10) subset during BC. Subjects performed three separate sets of 4, 8, and 12 repetitions for BC and KE at 65% one-repetition maximum. Rating of perceived exertion for the active muscles (RPE-AM) was measured during the mid and final repetition and RPE for the overall body (RPE-O) during the final repetition. : For both female and male groups across the three sets: (a) RPE-AM ranged from 3.6 to 8.2 for BC and 5.1 to 9.6 for KE and (b) RPE-O ranged from 2.4 to 6.7 for BC and 4.2 to 7.6 for KE. Positive linear regressions ranged from r = 0.79 to 0.91 ( P < 0.01) between Wttot and RPE-AM (mid), RPE-AM (final), and RPE-O for both BC and KE in both sex groupings. A positive ( P < 0.01) linear regression was found between [Hla] and RPE-AM (final) (r = 0.87) during BC. RPE did not differ between women and men at any measurement point within each set for BC and KE. RPE-AM (final) was greater ( P < 0.01) than RPE-O in the three sets of BC and KE. CONCLUSION: Findings provided concurrent validation of the OMNI-RES to measure RPE for the active muscle and overall body in young recreationally trained female and male weight lifters performing upper- and lower-body resistance exercise. PMID- 12569226 TI - Multicomponent cross-validation of minimum weight predictions for college wrestlers. AB - In 1998, the National Collegiate Athletic Association (NCAA) adopted a rule requiring that skinfolds (SF) or hydrostatic weighing (HW) be used to estimate minimum weight (MW) in college wrestlers. PURPOSE: To cross-validate the NCAA methods for estimation of MW using a multicomponent criterion (4C). METHODS: Criterion MW was calculated from body density (BD), bone mineral content (BMC), and total body water (TBW) using the 4C equation of Lohman (1992). BMC was measured by dual energy x-ray absorptiometry (DXA), TBW by deuterium dilution, and BD by HW. Subjects were Division I athletes from the University of Wisconsin (mean +/- SD; N = 33, age = 19.5 +/- 1.3 yr, height = 177.3 +/- 7.8 cm, weight = 74.2 +/- 9.3kg). RESULTS: There was no significant difference between mean MW from HW (69.6 +/- 8.5 kg) and SF (70.1 +/- 8.3 kg) ( P = 0.113), and between mean MW from HW (69.6 +/- 8.5 kg) and 4C (69.5 +/- 8.6 kg) ( P = 0.46). A clinically small, yet significant difference was seen when comparing mean MW from SF to 4C ( P = 0.013). The regression for the relationship between 4C and HW (y = 0.994 x HW + 0.294 kg, R2 = 0.985) and 4C and SF (y = 1.019 x SF - 1.885 kg, R2 = 0.979) did not significantly deviate from the line of identity. Pure error (PE) values of 1.04 kg and 1.35 kg were found for HW and SF, respectively. The difference between the methods was plotted as a function of the 4C criterion. The regression line for HW and 4C (y = -0.009x + 0.743, r = -0.07, P = 0.69) and SF and 4C (y = 0.038x + 3.259, r = -0.27, P = 0.13) suggest that no systematic differences in the prediction were associated with the size of the criterion. CONCLUSION: These data support the NCAA methods of HW and SF to predict MW when cross-validated using a 4C criterion in this sample. PMID- 12569227 TI - Impact of a competitive marathon race on systemic cytokine and neutrophil responses. AB - PURPOSE: To investigate whether cytokines and neutrophils mediate exercise related pathogenesis, we examined their responses and possible association after exhaustive exercise. METHODS: Plasma and urine samples were obtained from 10 male runners before and after a 42.195-km marathon race. Major cytokines and neutrophil activation markers [myeloperoxidase (MPO) and lactoferrin (LTF)] were measured by enzyme-linked immunosorbent assays. Functional modulation of standard neutrophils and monocytes by plasma was determined on their luminol-dependent chemiluminescence responses. RESULTS: The race induced peripheral neutrophilia accompanied by an increase in band neutrophils and monocytosis. Plasma MPO and LTF concentrations increased significantly by 1.8 and 1.4 times after the race. There was a greater increase in urine concentrations of MPO and LTF, 12.3 and 3.5 times after exercise, respectively, suggesting that neutrophil activation occurred and that renal clearance exceeded the increase in plasma concentrations. Plasma interleukin (IL)-6, IL-8, IL-10, granulocyte colony-stimulating factor (G CSF), macrophage CSF (M-CSF), and monocyte chemotactic protein 1 (MCP-1) increased significantly after the race, and urine IL-1beta, IL-6, G-CSF, M-CSF, and MCP-1 increased significantly. The plasma IL-6 responses correlated with the increases of band neutrophil count (r = 0.860, P < 0.01), suggesting IL-6 mediated bone marrow release of neutrophils. Furthermore, the increases in urine MPO concentration were correlated with increases in urine IL-6 (r = 0.868, P < 0.01) and G-CSF (r = 0.875, P < 0.01), suggesting that these cytokines promoted neutrophil activation. However, preincubation of neutrophils and monocytes with postexercise plasma could not cause priming responses, possibly because of the exercise-induced enhancement of plasma antioxidant activity. CONCLUSION: Although many cytokines recruiting and priming neutrophils and monocytes were secreted and functional after exhaustive exercise, overwhelming antioxidant and antiinflammatory defenses were induced, preventing exercise-induced oxidative stress. PMID- 12569228 TI - Effect of water polo practice on cytokines, growth mediators, and leukocytes in girls. AB - PURPOSE: The effects of exercise on growth and development are mediated through a complex interaction between the endocrine, immune, and nervous systems. Very little is known about how these systems respond to exercise in children or adolescents. Moreover, there are few studies that have examined growth factors, inflammatory cytokines, and leukocyte responses to "real-life" or field exercise solely in girls. Thus, the goal of the present study was to determine the acute exercise-induced alterations in the growth hormone --> insulin-like growth factor I axis, inflammatory cytokines, and certain aspects of immune function in a group of adolescent girls after a typical water polo practice. METHODS: Ten, healthy, high-school female subjects, 14-16 yr old, performed a single, typical, 1.5-h water polo practice session. Blood was sampled before and after the session. RESULTS: The exercise resulted in an increase in HR (from 82 +/- 2 to 161 +/- 5 beats.min(-1) at 30 min, P < 1.4.10(-6) ), as well as in circulating lactate levels (375 +/- 66%, P < 0.0005). Significant increases where noted in circulating IL-6 (396 +/- 162%, P < 0.005) and IL-1ra (71 +/- 20%, P < 0.015). A substantial increase in the level of IGFBP-1 (1344 +/- 344%, P < 0.001) was also observed. Interestingly, TNF-alpha levels decreased after the exercise (-10.4 +/- 3.8%, P < 0.04) as did insulin (55 +/- 12%, P < 0.005). The exercise led to significant increases in granulocytes, monocytes, and lymphocytes. The exercise significantly influenced adhesion molecules (such as CD62L and CD54), which has not been previously studied in adolescent girls. CONCLUSIONS: These data demonstrate that an intense "real-life" exercise bout in adolescent females leads to profound increases in inflammatory cytokines and reductions in anabolic mediators with substantial alterations in white blood cell subpopulations and adhesion molecules. The role of these frequent, almost daily immune and cytokine changes on growth and development have yet to be determined. PMID- 12569229 TI - Physical demands of vacuuming in women using different models of vacuum cleaners. AB - PURPOSE: Women continue to have the primary responsibility for housekeeping, even after a cardiac event. Vacuuming is one housekeeping task that is often reported as difficult to perform due to angina symptoms. The aim of this study was to evaluate the energy expenditure and hemodynamic responses associated with vacuuming using five different models of vacuum cleaners. METHODS: Thirty-six healthy women 50-59 yr of age (54.5 +/- 3.1 yr) participated in this study. Energy expenditure was measured by indirect calorimetry using an Aerosport KB1-C portable metabolic system. After collection of baseline HR, blood pressure (BP), and oxygen consumption ( VO2 ) data, subjects performed vacuuming and treadmill walking in one of six different sequences. Vacuuming consisted of using five different models of vacuum cleaners, three upright models (heavy-duty, self propelled, and lightweight) and two canister models (standard and compact) for 6 min each. Treadmill walking was conducted at 2.0 mph (0% grade) for 6 min. VO2, HR, BP, and RPE were recorded during each task. The rate-pressure product (RPP) was calculated to estimate myocardial oxygen demand. RESULTS: Vacuuming with the self-propelled upright model resulted in significantly lower VO2, RPE, HR, systolic BP, and RPP responses compared with some of the other models.(2) CONCLUSION: Significant differences in oxygen consumption and myocardial oxygen demand are associated with vacuuming using different models of vacuum cleaners. When making recommendations to individuals regarding the least physiologically demanding models, the power assist features of the machine should be the greatest consideration. This important characteristic should be taken into account when making recommendations for patients with limited capacity, or those wanting to limit physiological stress due to a disease state. PMID- 12569230 TI - Alcohol and cardiovascular disease--more than one paradox to consider. Type of alcoholic beverage and cardiovascular disease--does it matter? AB - While a large number of studies have shown a U- or J-shaped relation between alcohol intake and cardiovascular disease, the question remains whether this risk function persists in all subsets of the population or derives from all types of alcoholic beverages. Recent large population studies have suggested that the effect of alcohol and cardiovascular disease and all cause mortality is modified by factors such as age, gender, cardiovascular risk factor status, drinking pattern and type of alcohol. The author reviews the studies indicating this effect modification, while focusing on the potential beverage specific differences. The review goes through several of the methodological issues in studying the effects of the different types of beverages on health, and gives some of the plausible biological mechanisms that might explain the differences, but stresses the different biases, which are the reasons why the issue is not settled yet. PMID- 12569231 TI - Alcohol and cardiovascular disease--more than one paradox to consider. Alcohol and hypertension--does it matter? (no!). AB - Good evidence suggests that alcohol probably has a causal relationship to hypertension, although many possible confounding factors that may exaggerate or attenuate the relationship, if true. Alcohol can also adversely affect other systems, including the heart (arrhythmias, alcoholic cardiomyopathy, etc.), the liver (alcoholic hepatitis, cirrhosis, etc.) and the nervous system (peripheral neuropathy, etc.). Hypertension is very common and it is unlikely that all (or most) of hypertensives can identify alcohol as causative. Indeed, hypertension is likely to be multifactorial and many factors would confound the relationship, if any, between alcohol and hypertension. PMID- 12569232 TI - Alcohol and cardiovascular disease--more than one paradox to consider. Average volume of alcohol consumption, patterns of drinking and risk of coronary heart disease--a review. AB - The effect of average volume of alcohol on coronary heart disease (CHD) is J shaped in established market economies. Light to moderate drinkers have less risk than abstainers, with heavy drinkers displaying the highest level of risk. This relationship between average volume of alcohol consumption and CHD is modified by different patterns of drinking. Heavy drinking occasions as well as drinking outside meals are related to increased CHD risk, independently of volume of drinking. Beverage type does not seem to have much impact, even though there are some indications that wine is more protective than other forms of alcohol. Physiological mechanisms have been identified to explain this complex relationship between alcohol and CHD. Since patterns of drinking are important in determining CHD risk, they should be included in future epidemiologic studies, together with biomarkers further to test hypotheses about pathways. PMID- 12569233 TI - Alcohol and cardiovascular disease--more than one paradox to consider. Alcohol and hypertension: does it matter? Yes. AB - Observational studies and clinical experiments solidly establish an empiric relationship between heavy drinking (>/= 3 standard drinks/day) and higher blood pressure. A biologic mechanism for this link remains unclear. It has been difficult to study the incidence of usual hypertension sequelae in alcohol associated hypertension because of complex interactions of alcohol, hypertension and cardiovascular conditions. Even with these limitations, a causal association is the most likely explanation and control of alcohol intake is probably important in prevention and treatment of hypertension. PMID- 12569234 TI - Alcohol and cardiovascular disease--more than one paradox to consider. Alcohol and type 2 diabetes--another paradox? AB - The French Paradox relates to the observation that mortality rates due to coronary heart disease are relatively low in France despite a diet rich in saturated fats. Another paradox linked to alcohol is the diverse associations of acute and chronic alcohol use with respect to insulin resistance, incidence of type 2 diabetes and incidence of cardiovascular disease in type 2 diabetes. Reports consistently suggest that the acute affects of alcohol induce a state of insulin resistance following either an oral and/or intravenous glucose load. Contrary to the acute alcohol studies is a large body of epidemiological evidence from cross-sectional studies which suggests that long-term exposure to alcohol is associated with an improvement in insulin sensitivity. Furthermore, a substantial number of prospective studies point to a protective role for light to moderate chronic alcohol intake against the development of diabetes as well as a protective effect of regular mild to moderate drinking against coronary artery disease in type 2 diabetic subjects. PMID- 12569236 TI - Novel biomedical risk markers for cardiovascular disease. AB - Molecular moieties within newly discovered pathophysiological pathways in atherothrombosis represent potential biomedical risk markers for cardiovascular disease. Their discovery is important in that a large proportion of cardiovascular disease is unexplained by 'conventional' risk markers, and that novel markers may pave the way to new targets for intervention. Many candidate markers have been identified, among which inflammatory and haemostatic factors are prominent. This review will examine the evidence for the roles of key novel risk markers in cardiovascular disease, against criteria which define causality as well as potential clinical utility. PMID- 12569235 TI - Effects of a general practice-based intervention on diet, body mass index and blood lipids in patients at cardiovascular risk. AB - BACKGROUND: General practice-based health-promotion programmes implemented by nurse-counsellors may reduce cardiovascular risk factors, particularly in patients at increased risk. DESIGN: In a randomized controlled trial, trained nurse-counsellors delivered cognitive behavioural programmes aiming to reduce cardiovascular risk in patients with hypertension, Type 2 diabetes mellitus or coronary heart disease. Of 591 eligible patients from seven general practices in Perth, Western Australia, 212 agreed to participate and were randomized to one of three groups. In the 'Low' intervention (n = 69), monthly telephone contacts for 1 year followed one face-to-face individual counselling session; in the 'High' intervention (n = 74) individual face-to-face counselling continued over 1 year, taking place monthly, for up to 1 h; Controls (n = 69) received usual care only. Participants were assessed at baseline with follow-up 12 and 18 months later. RESULTS: Follow-up assessments were attended by 77, 71 and 65% of the Control, 'Low' and 'High' groups, respectively. Total fat intake fell by 9, 12 and 5% in the 'High', 'Low' and Control groups, respectively, at 12 months and by 2, 10 and 5% at 18 months. Total serum cholesterol fell by 3, 3 and 2% in the 'High', 'Low' and 'Control' groups, respectively, at 12 months and by 7, 5 and 8% at 18 months. Neither change in fat intake nor serum cholesterol differed significantly between groups. Body mass index increased in all groups with no significant changes related to the programme. CONCLUSIONS: Programmes using nurse-counsellors were not significantly more successful than usual care from general practitioners, consistent with reported benefits of lifestyle advice from primary-care physicians. PMID- 12569237 TI - Influence of hormone replacement therapy on C-reactive protein: population-based data. AB - BACKGROUND: Observational data, possibly resulting from confounding due to self selection, show that use of hormone replacement therapy (HRT) is associated with reduced coronary risk, while randomized trial data do not. One possible explanation for the trial findings is that adverse effects of HRT-- such as an elevation of C-reactive protein (CRP)--might counterbalance other benefits of HRT. DESIGN: Women aged 40-74 years were identified from a cross-sectional, annual nationally representative study of the English population, the Health Survey for England 1998. Valid data were available from 4,112 of the 4,319 women in this age group. Of these, 710 (17%) were current, 465 (11%) past and 2,937 (71%) never users of HRT. METHODS: To study the association between current and past use of HRT and CRP levels. CRP was expressed as a continuous variable (logged) and in percentiles. RESULTS: Current, past and never users of HRT did not differ in terms of age, body mass index or smoking status. However, compared with past and never users of HRT, current users tended to take more vigorous physical activity and were more likely to drink more alcohol than is currently recommended. Excluding potentially confounding conditions, median CRP levels were significantly higher in current (2.5 mg/l) and in past (1.9 mg/l) than never HRT users (1.4 mg/l). Controlling for age, smoking and body mass index logged CRP remained significantly raised only among current users. CONCLUSIONS: HRT use was significantly and independently associated with raised CRP levels in an English nationally representative sample. Increases in CRP may represent one plausible mechanism by which HRT may adversely affect risk of coronary heart disease. PMID- 12569238 TI - Insufficient use of lipid-lowering drugs and measurement of serum cholesterol among patients with a history of myocardial infarction. AB - BACKGROUND: The effect of lipid-lowering drugs (LLDs) on coronary heart disease is well documented, particularly in patients with established ischaemic heart disease. However, intensity of the use of these drugs is low. The aim of this linkage study was to analyse the use of serum cholesterol measurements and LLDs among patients with a history of myocardial infarction (MI) in a Danish population. METHODS: Information on serum cholesterol values was retrieved from the hospital's Department of Clinical Chemistry for all patients from the Odense municipality and four surrounding municipalities (213,868 inhabitants) hospitalised at Odense University Hospital for MI between 1994-1997. Information on LLD use was obtained from a prescription database. Only patients alive at discharge were included in the investigation. The total observation period was from 1993-1998. RESULTS: In all, 1,018 patients were eligible for observation and 39% of the patients who met the criteria for LLD reimbursement (MI and serum cholesterol >5.4 mmol/L) started treatment. Relatively more males than females and relatively more patients under the age of 70 years were treated. Patients above the age of 70 years were prescribed these drugs at a slower rate. CONCLUSIONS: More attention to the insufficient use of lipid-lowering drug treatment in patients with established coronary heart disease is needed. PMID- 12569239 TI - Cardiovascular risk factors in the elderly: the Tehran Lipid and Glucose Study. AB - BACKGROUND: Coronary artery disease is becoming more prevalent in developing countries, particularly in urban areas. Because the proportion of elderly individuals in the population is on the rise, this study was conducted to determine the prevalence of cardiovascular risk factors among the Tehran urban elderly population. DESIGN AND METHODS: Among 15,005 urban individuals of 3 years old and over who had been chosen in a cross-sectional phase of a longitudinal study in Tehran, there were 1,799 people aged 60 years and over. The prevalence and distribution of high blood pressure, cigarette smoking, dyslipoproteinemia, diabetes mellitus and obesity were determined in this population. Dietary intake was assessed in a subsidiary of 54 people by means of two 24 h dietary recalls. RESULTS: The percentage of women with two or more cardiovascular disease risk factors was significantly greater than in men (74% compared with 53%, P < 0.001). One fourth of men and 55% of women had high serum cholesterol levels (>/=240 mg/dl). The prevalence of diabetes mellitus and impaired glucose tolerance was 24% and 21% in men and 29% and 20% in women, respectively. The prevalence of obesity (body mass index >/=30 kg/m(2)) was 15% for men and 36% for women. Fifty five per cent of men and 94% of women had high waist-to-hip ratios (>0.95 in men and >0.8 in women). The mean percentage values of energy intake derived from carbohydrate, protein and fat were 60.5 +/- 8.0, 11.5 +/- 2.0 and 27.8 +/- 8.9, respectively. CONCLUSIONS: The prevalence of cardiovascular risk factors among the Tehran urban elderly population is high. Some efforts should be made to reverse the recent trend towards increasing age-related mortality and morbidity rates of coronary heart disease. PMID- 12569240 TI - The ALLHAT trial: a verdict or a challenge? PMID- 12569241 TI - All hats off to ALLHAT: a massive study with clear messages. PMID- 12569242 TI - The ALLHAT trial: strengths and limitations. PMID- 12569243 TI - The meaning of ALLHAT. PMID- 12569244 TI - Comments on the renal aspects of the ALLHAT study. PMID- 12569245 TI - Influence of body and arm position on blood pressure readings: an overview. PMID- 12569246 TI - Publication bias and meta analysis. PMID- 12569247 TI - Problems with linkage analysis and QTL detection in hypertension. PMID- 12569248 TI - Salt sensitivity and arterial structure and function. PMID- 12569249 TI - Intracellular Ca2+ and renin gene transcription. PMID- 12569250 TI - Inhibitors of Na+/Mg2+ exchange activity attenuate the development of hypertension in angiotensin II-induced hypertensive rats. PMID- 12569251 TI - Anti-atherosclerotic effects by AT1 receptor blockade. PMID- 12569252 TI - On a low calorie diet, are there separate and discrete effects of negative energy balance and weight loss on blood pressure? PMID- 12569253 TI - Reactive oxygen species and pre-eclampsia. PMID- 12569254 TI - Essential hypertension and left ventricular hypertrophy in cardiovascular disease: beyond overactivity of the renin-angiotensin-aldosterone-sodium system. PMID- 12569255 TI - Altered tubular albumin degradation in the pathogenesis of albuminuria: additional experimental evidence challenging the paradigm of glomerular capillary leakage. PMID- 12569256 TI - Publication bias and heterogeneity in the relationship between systolic blood pressure, birth weight, and catch-up growth--a meta analysis. AB - OBJECTIVE: To use meta-analytic techniques to formally examine (1) publication bias, and (2) between-study heterogeneity in the reported regression coefficient of systolic blood pressure (SBP) on birth weight, adjusting for current weight. DATA IDENTIFICATION: A total of 55 separate analyses reporting on 382 514 male and female subjects ranging from 0 to 75 years, summarized in a recent systematic review. RESULTS: Some 52 of 55 analyses reported a negative regression coefficient (beta) of SBP on birth weight, adjusting for current size, and in 27 of 55 (49%) the 95% confidence interval (CI) did not include zero. A pooled estimate of the regression coefficient across studies under a random-effects model was -1.38 mmHg/kg (95% CI: -1.66 to -1.10). The magnitude of the regression coefficient was negatively correlated with the age of the study population (r = 0.44, P = 0.0007). However, there was strong evidence of publication bias with smaller studies tending to report more negative betas - the Pearson correlation between beta and its standard error (SE(beta)) was -0.55 (P < 0.0001). To examine independent effects, meta analysis regressions of beta on age and SE(beta) were run. After controlling for age, there remained strong evidence of publication bias (P = 0.002), whereas after controlling for publication bias as measured by SE(beta), the relationship between beta and age was not significant (P = 0.08). CONCLUSIONS: This analysis strongly suggests, but does not prove, that publication bias is present when systematically examining the relationship between blood pressure and birth weight. Controlling for publication bias lessens the apparent relationship between the regression coefficient and age of the study population. PMID- 12569257 TI - Serum lipoprotein(a) concentrations and alcohol consumption in hypertension: possible relevance for cardiovascular damage. AB - OBJECTIVES: To evaluate the relationships between alcohol intake and serum lipoprotein(a) [Lp(a)], a powerful predictor of organ damage, in patients with essential hypertension with a wide range of alcohol intake, and to investigate whether the association between alcohol intake and serum Lp(a) concentrations occurs over the entire spectrum of apo(a) phenotypes. DESIGN: Cross-sectional study of a case series. SETTING: University medical centre. PATIENTS: Four hundred and two patients with untreated essential hypertension recruited at a hypertension clinic. MAIN OUTCOME MEASURES: Serum Lp(a) concentrations, apo(a) isoforms, alcohol consumption, smoking habits and cardiovascular status. RESULTS: No difference in Lp(a) concentrations was observed between teetotalers and occasional drinkers. Light drinkers (1-20 g/day ethanol), moderate drinkers (21 50 g/day), and heavy drinkers (> 50 g/day) had, respectively, 21, 26 and 57% lower median Lp(a) concentrations than teetotalers and occasional drinkers. Similar findings were obtained when male and female patients were analysed separately. Log Lp(a) concentrations were inversely and independently correlated with alcohol consumption in both men and women with hypertension. The frequency distributions of apo(a) isoforms and liver function parameters were comparable across the different alcohol intake groups. Patients with evidence of cardiovascular damage had greater concentrations of serum Lp(a) and higher frequency of low-molecular weight apo(a) isoforms as compared with patients without such evidence. CONCLUSIONS: Serum Lp(a) is inversely and dose-dependently related with alcohol intake in patients with hypertension, and this relationship is independent of the size distribution of apo(a) isoforms. Reduction of Lp(a) concentrations by regular consumption of alcohol might favourably affect the atherosclerotic risk profile of patients with hypertension and thereby decrease cardiovascular morbidity. PMID- 12569258 TI - Sodium intake and blood pressure in healthy individuals. AB - OBJECTIVE: To investigate further the relationships between blood pressure and sodium intake, which have been claimed to exist, but have not been clearly established, in epidemiological studies. DESIGN AND PARTICIPANTS: A 2-year follow up study of a cohort of 296 healthy volunteers working in the same company. RESULTS: Consistent with previously reported findings, covariance analysis in the entire population, including multivariate analysis to examine the potential influence of confounding variables, failed to identify a significant relationship between blood pressure and sodium intake or between their respective changes within 2 years. To test the hypothesis of a heterogeneous distribution of salt sensitivity, we calculated the statistical dependence between blood pressure and sodium intake using the coefficient analysis that quantifies the prevalence of a statistical link between two variables. A significant dependence (P < 0.05) between diastolic (DBP) or systolic (SBP) blood pressure and sodium intake was found in 16 and 5% of the participants, respectively. The analysis of the 2-year changes in blood pressure and sodium intake reinforced our findings. Our finding of a relationship between blood pressure and sodium intake in only a few individuals may explain the lack of a global relationship between these two variables investigated by the covariance analysis in our study.(Rho) CONCLUSION: Our results suggest that 5-16% of healthy individuals have a 'salt-dependent blood pressure' and might benefit from a reduction in dietary salt intake. PMID- 12569259 TI - Genome-wide searches for blood pressure quantitative trait loci in the stroke prone spontaneously hypertensive rat of a Japanese colony. AB - OBJECTIVES: Although several quantitative trait loci for blood pressure have been reported in stroke-prone spontaneously hypertensive rats (SHRSP), the results are not always concordant among different crosses. To evaluate potential confounding factors in linkage analysis, we performed genome-wide screens in F2 populations derived from SHRSP and Wistar-Kyoto rats of a Japanese colony. METHODS: Two F cohorts were independently produced: F2-1 (110 male and 110 female rats), and F2 2 (174 male and 184 female rats). Blood pressure was measured longitudinally (from 2 to 5 months of age and 1 month after salt-loading) in F2-1, while it was measured at 13 weeks of age in F2-2. Subsequent to an initial screen with 251 markers in F2-1 male progeny, 170 markers were selected and characterized in the remaining populations. RESULTS: When 578 rats were analyzed together, markers from five chromosomal regions showed significant linkage to blood pressure at 13 weeks of age. The strongest and the most consistent linkage was found on rat chromosome 1 (a maximal log of the odds score reached 8.3). In the other regions, the degree of linkage was more prominent in either of sexes. Some evidence of age specific and sex-specific linkage was detected in five additional regions in the F2-1 cohort. In the Japanese colony, however, there was no significant linkage to several chromosomal regions previously reported in other SHRSP colonies. CONCLUSIONS: Our data provide solid evidence of a chromosome-1 linkage and demonstrate the importance of aging, sex, and dietary manipulation in linkage analysis. Also, the combination of parental rat strains seems to be critical when searching for blood pressure quantitative trait loci. PMID- 12569260 TI - Linkage analysis of 2q14-q23 and 5q32 with blood pressure quantitative traits in Chinese sib pairs. AB - OBJECTIVES: Several genome-wide scans recently accomplished in the ethnic Chinese revealed a number of candidate loci possibly contributing to essential hypertension, and some appeared to be replicable in 2q14-q23 and 5q32. The current study aimed to examine the linkage of qualitative and blood pressure quantitative traits in essential hypertension with these genomic regions in a large sample of Chinese hypertensive families. METHODS: We performed a genetic analysis on 148 randomly ascertained families containing 328 affected sib pairs, grouped into two geographically distinct subsets. Five highly informative microsatellite markers (D2S151, D2S142, D5S2090, D5S413 and D5S2013) were genotyped, and linkage analyses were performed with different genetic models. RESULTS: We did not observe consistent evidence for excess allele sharing identity by descent in either of the qualitative or the quantitative test. However, higher LOD scores were found at D5S2013 in North Group subset with Haseman-Elston and maximum likelihood (ML) variance (no dominance variance, NDV) algorithms. With the ML (NDV) algorithm, the LOD was 1.410 for diastolic blood pressure at this locus, although this was not statistically significant. CONCLUSIONS: These findings provide no evidence to support a significant linkage of 2q14-q23 or 5q32 with essential hypertension or blood pressure quantitative traits in the ethnic Chinese, and indicate the aetiologic diversity and complexity of hypertension. Previous reports implied 2q14-q23 or beta 2- adrenergic receptor gene potentially linked to essential hypertension in the ethnic Chinese. To replicate these results and perform quantitative linkage analysis, we genotyped members of 148 hypertensive families with five highly informative microsatellite markers. We observed no evidence of excess allele sharing identity by descent in sib pairs, revealing a lack of linkage between 2q14-q23 or 5q32 (chromosome region harboring the gene encoding beta 2 adrenergic receptor) and hypertension in our study sample. PMID- 12569261 TI - Altered structure and reduced distensibility of arteries in Dahl salt-sensitive rats. AB - OBJECTIVES: The role of salt sensitivity in arterial stiffening and the structural basis of reduced arterial distensibility were investigated in Dahl salt-sensitive (DS) rats. METHODS: Three-month-old male DS rats received a normal (0.7% NaCl) or a high-sodium (2% NaCl) diet for 3 months. Dahl salt-resistant (DR) rats were controls. Pressure-volume (distensibility) relationships were measured in in-vitro-perfused segments of right carotid and iliac arteries, in the presence and absence of extracellular calcium. The left carotid and iliac arteries were perfusion-fixed at 100 mmHg for morphometric measurements. RESULTS: The average monthly tail systolic blood pressure (SBP) of DS rats on normal and high-sodium diets were increased compared to that of DR rats. Compared to controls, carotid and iliac artery pressure-volume curves of DS rats on normal and high-sodium diets were shifted toward the pressure axis, without a change in elastic moduli. In DS rats, reduced distensibility of the carotid artery was accompanied by increased lumen diameter and increased thickness of media and elastic lamellae, the wall to lumen ratio being unchanged; wall thickness was increased and lumen diameter unchanged in the iliac artery. The high-sodium diet had no effect on either distensibility or dimensions of carotid and iliac arteries in DS or DR rats. CONCLUSION: Geometry (increased or unchanged lumen and increased wall thickness), rather than increased stiffness of wall components, appears to be the cause of reduced distensibility of arteries in DS rats. Structural and functional adaptation to salt sensitivity may occur on what is considered a 'normal' sodium diet. PMID- 12569262 TI - Adolescent blood pressure does not predict aortic stiffness in healthy young adults. The Atherosclerosis Risk in Young Adults (ARYA) study. AB - BACKGROUND: Increased arterial stiffness has been shown to be a good predictor of cardiovascular morbidity and mortality and to be associated with an adverse cardiovascular risk profile. OBJECTIVE: To evaluate the relationship between adolescent blood pressure and aortic stiffness in 524 healthy young adults aged 27-30 years, as a means of investigating early determinants of arterial stiffness. SETTING: General community. PARTICIPANTS: We studied 524 healthy young adults, aged 27-30 years, who attended secondary school in Utrecht, The Netherlands. MEASUREMENTS: Data on adolescent weight, height, blood pressure and stage of puberty were available from the original school health records of the Municipal Health Service. At young adulthood, a questionnaire on cardiovascular risk factors was completed and a fasting blood sample was drawn. MAIN OUTCOME MEASURE: Arterial stiffness, assessed by carotid-femoral pulse wave velocity (PWV). RESULTS: Adolescent blood pressure did not predict aortic PWV at young adulthood (linear regression coefficient 0.03 m/s per 10 mmHg increase in systolic blood pressure; 95% confidence interval 0.09 to 0.14). Repeated analysis in a subgroup (n = 199) for whom two adolescent blood pressure measurements were averaged showed stronger (2-19x) associations with adult PWV, although these were not significant. CONCLUSION: Adolescent blood pressure did not predict arterial stiffness in healthy young adults. Measurement error in the baseline blood readings (regression to the mean phenomenon) may partly explain the lack of association in our study. Further studies should confirm our results in order to enable better understanding of the role of adolescent blood pressure in the aetiology of vascular damage. PMID- 12569263 TI - Implication of Ref-1 in the repression of renin gene transcription by intracellular calcium. AB - OBJECTIVE: The production of renin, which catalyzes the rate-limiting step of the renin-angiotensin system, is tightly regulated by intracellular second messengers. Among them, an increase of intracellular calcium represses renin gene expression. This inhibition of gene expression by intracellular calcium is exceptional, and the molecular mechanism supporting this phenomenon has not yet been identified. As the renin gene is negatively regulated by calcium in the same way as the parathormone (PTH) gene, we hypothesized that a similar molecular transcriptional mechanism could be involved. RESULTS: Analysis of the human renin proximal promoter led to the identification of a negative calcium response element (nCaRE), which is identical to the region of the PTH promoter and is involved in its repression by calcium. Transfection experiments in renin expressing chorio-decidual cells demonstrated the transcriptional functionality of the human renin promoter nCaRE. In addition, mutation of nCaRE suppressed the sensitivity of the renin promoter to the increase in intracellular calcium. Gel shift assays demonstrated that Redox factor 1, a multifunctional protein involved in the repair of damaged DNA and the redox activation of AP-1 transcriptional factors, binds specifically to nCaRE. Immunostaining showed that this factor is translocated from the cytoplasm to the nucleus in response to an increase in the intracellular calcium concentration. CONCLUSION: Thus, the repression of renin expression by intracellular calcium may be mediated by the calcium-induced translocation of Ref-1 to the nucleus, where it binds to the renin promoter nCaRE, to repress the transcription of the renin gene. PMID- 12569264 TI - Inhibitors of Na+/Mg2+ exchange activity attenuate the development of hypertension in angiotensin II-induced hypertensive rats. AB - OBJECTIVES: To investigate whether imipramine and quinidine, inhibitors of the Na /Mg exchanger, influence development of hypertension in rats infused with angiotensin (Ang) II. METHODS: Sprague-Dawley rats were divided into six groups: (1) control (vehicle); (2) Ang II (150 ng/kg per min subcutaneously); (3) imipramine alone (5 mg/kg per day in drinking water); (4) quinidine alone (5 mg/kg per day in drinking water); (5) Ang II plus imipramine; (6) Ang II plus quinidine. Rats were studied for 3 weeks. To verify that Ang II directly influences Na -dependent Mg exchange, in-vitro studies were performed in vascular smooth muscle cells (VSMCs) derived from mesenteric arteries. RESULTS: Ang II increased systolic blood pressure (SBP) in all groups. The magnitude of the increase was lower ( 0.01) in Ang II groups treated with imipramine (151 +/- 7.4 mmHg) or quinidine (163 +/- 4 mmHg) than in the Ang II only group (205 +/- 4 mmHg). Neither imipramine nor quinidine influenced SBP in vehicle-treated rats. Plasma concentrations of Mg and K were decreased in Ang II rats compared with controls (P < 0.05). Platelet intracellular free Mg concentration was reduced and platelet intracellular free Na concentration was increased in the Ang II group compared with control and treated groups (P < 0.01). These effects were normalized by imipramine and quinidine. Ang II stimulated Na -dependent Mg transport in VSMCs. These actions were abrogated by imipramine and quinidine and in Na -free conditions. CONCLUSIONS: Our data demonstrate that inhibitors of Na dependent Mg transport attenuate development of hypertension in rats infused with Ang II. These findings suggest a possible role for Na /Mg exchange activity in the pathogenesis of Ang II-dependent hypertension. PMID- 12569265 TI - Role of angiotensin II in L-NAME-induced systemic and renal hemodynamic effects in hydrochlorothiazide-pretreated hypertensive subjects. AB - BACKGROUND: Experimental evidence suggests that in conditions associated with an activated renin-angiotensin system, unopposed activity of angiotensin II underlies the marked renal vasoconstrictor response to nitric oxide synthase inhibition. In the present study, we investigated whether this holds true in hypertensive subjects pretreated with hydrochlorothiazide (HCT). METHODS: Systemic N(G)-nitro-L-arginine methyl ester (L-NAME) infusions (12.5 microg/kg per min for 40 min) were given to eight hypertensive subjects (age 53 +/- 6 years) during placebo, and during pretreatment with HCT (25 mg once daily) or HCT and losartan (LOS) (50 mg twice daily), both for 9 days. The glomerular filtration rate (GFR) and renal plasma flow were estimated from the clearances of radiolabeled thalamate and hippuran. Renal blood flow (RBF) was calculated as renal plasma flow/1 - hematocrit and the renal vascular resistance (RVR) as mean arterial pressure (MAP) divided by RBF. RESULTS: Compared with placebo, plasma renin increased (P < 0.001) from 15 +/- 4 mU/l during placebo to 26 +/- 7 mU/l during HCT and to 133 +/- 51 mU/l during HCT + LOS. MAP (110 +/- 3 mmHg) decreased to 102 +/- 4 mmHg during HCT and to 98 +/- 5 mmHg during HCT + LOS. RBF (579 +/- 36 ml/min), GFR (97 +/- 6 ml/min) and filtration fraction (29 +/- 2%) did not change, whereas RVR (200 +/- 15 RU) decreased to 183 +/- 13 RU during HCT and to 165 +/- 14 RU during HCT + LOS (P < 0.05). In response to L-NAME, MAP and RVR increased maximally by 10 +/- 3 and 67 +/- 9%, whereas RBF and GFR decreased maximally by 42 +/- 6 and 18 +/- 4%. Compared with these responses, the responses of MAP, RBF and RVR were not affected by pretreatment of HCT or HCT + LOS, but the L-NAME-induced decrease in GFR (26 +/- 5% during HCT and 29 +/- 5% during HCT and LOS) was enhanced (P < 0.01). CONCLUSIONS: In hypertensive subjects with an activated renin-angiotensin system, unopposed activity of angiotensin II is not involved in L-NAME-induced pressor and renal vasoconstrictor response, whereas the L-NAME-induced decrease in GFR is enhanced, indicating greater dependency of GFR on nitric oxide-mediated vasodilator tone during sodium depletion. PMID- 12569266 TI - Proximal tubular fluid angiotensin II levels in angiotensin II-induced hypertensive rats. AB - BACKGROUND: It has been shown that infusions of low-dose angiotensin II (Ang II) for 2 weeks lead to impaired pressure natriuresis and autoregulatory capability. Although intrarenal renin content and renin mRNA levels are markedly reduced, whole-kidney Ang II content has been shown to be increased. However, the intrarenal distribution of the increased intrarenal Ang II has not been established. OBJECTIVE: To determine the concentrations of Ang II in the proximal tubule fluid achieved in hypertensive rats (n = 16) infused with Ang II, previously prepared by infusion with Ang II at 60 ng/min via osmotic minipump for 13 days. METHODS: Rats were anesthetized with pentobarbital sodium and prepared for micropuncture, and then several free-flow proximal tubular fluid collections were obtained and pooled for each rat. At the end of each experiment, a blood sample was collected and the micropunctured kidney was excised and homogenized in chilled methanol. All samples were extracted immediately after collection and stored at 20 degrees C until the day of Ang II radioimmunoassay. RESULTS: Mean arterial blood pressure averaged 179 +/- 3 mmHg, renal plasma flow was 1.89 +/- 0.15 ml/min per g, and glomerular filtration rate averaged 0.58 +/- 0.04 ml/min per g. The Ang II concentration in proximal tubular fluid averaged 4.5 +/- 1.1 pmol/ml, a value substantially greater than the Ang II concentrations in plasma (0.17 +/- 0.03 pmol/ml), urine (0.06 +/- 0.01 pmol/ml), or total kidney tissue (0.40 +/- 0.10 pmol/g). Plasma renin activity (1.0 +/- 0.21 ng Ang I/ml per h) was markedly suppressed, as observed previously. CONCLUSIONS These findings indicate that Ang II concentrations in proximal tubular fluid collected from kidneys of anesthetized hypertensive rats infused with Ang II are in the nanomolar range, similar to those observed in normotensive rats. The inappropriate maintenance of nanomolar concentrations of Ang II in proximal tubular fluid of Ang II-infused hypertensive rats, even at markedly increased arterial pressures, may contribute to the impaired pressure natriuresis capability previously reported and, thereby, to the development and maintenance of hypertension in this model. PMID- 12569267 TI - Mechanisms of angiotensin II type 1 receptor blocker for anti-atherosclerotic effect in monkeys fed a high-cholesterol diet. AB - OBJECTIVE To clarify the mechanism of the anti-atherosclerotic effect of angiotensin II type 1 receptor blocker (ARB) in primates, we investigated whether an ARB (CS-866) affects the serum markers of inflammation and growth factors, and the endothelial function in monkeys fed a high-cholesterol diet. DESIGN Monkeys fed a high-cholesterol diet for 6 months were divided into two groups: one group was given an ARB, CS-866 (10 mg/kg per day), and the other group was not. The control group was fed a normal diet. RESULTS Blood pressure and the plasma cholesterol level were not affected by CS-866. Plasma levels of angiotensin II, renin, angiotensin converting enzyme and chymase were not changed by the high cholesterol diet, whereas vascular angiotensin converting enzyme, but not chymase, was significantly increased. Serum levels of macrophage-colony stimulating factor, transforming growth factor-beta1 and intracellular adhesion molecule-1 were significantly increased in monkeys fed a high-cholesterol diet but they were suppressed by CS-866. The relaxation response of isolated carotid arteries to acetylcholine was suppressed in the high-cholesterol group, whereas it was improved by CS-866. CONCLUSIONS CS-866 reduced lipid deposition along with the suppression of serum macrophage-colony stimulating factor, transforming growth factor-beta 1 and intracellular adhesion molecule-1, and the improvement of vascular functions, suggesting that ARB has multiple mechanisms for reducing lipid deposition in primates. PMID- 12569268 TI - Weight loss and weight maintenance, ambulatory blood pressure and cardiac autonomic tone in obese persons with the metabolic syndrome. AB - BACKGROUND: Cardiac autonomic function may play a role in obesity-associated hypertension. Most studies on the effects of weight loss on blood pressure and autonomic function do not distinguish between acute or continuing weight loss and steady-state weight maintenance after weight loss. OBJECTIVES: We sought to clarify the changes in ambulatory blood pressure, heart rate and heart rate variability as assessed by spectral analysis during rapid weight loss and extended weight maintenance. PARTICIPANTS: Abdominally obese (body mass index 35.2 +/- 2.1 kg/m2, waist 114.3 +/- 9.0 cm) men and women (n = 41) with the metabolic syndrome. METHODS AND RESULTS: The 34 men and women completing the 1 year weight maintenance period lost 14.6 +/- 3.5 kg during the 9-week very-low calorie diet and maintained a 12.5 +/- 7.5 kg weight loss to the end of the trial. Ambulatory 24-h blood pressure decreased dramatically during the diet ( 9.0 +/- 8.0/-4.6 +/- 4.9 mmHg), but had risen to near baseline levels by the end of weight maintenance (-2.2 +/- 8.2 /-1.2 +/- 6.1 mmHg). Night-time heart rate decreased (-5.5 +/- 9.6 beats/min, P = 0.012), and heart rate variability total and low-frequency power measured during 5 min of controlled breathing increased by 46-56% (P = 0.003-0.09) during rapid weight loss. These changes gradually attenuated during weight maintenance, and only the change in night-time heart rate was still of borderline significance after 1 year of weight maintenance ( 3.6 +/- 8.6 beats/min, P = 0.063). Heart rate variability high-frequency power tended to increase during weight loss and weight maintenance. CONCLUSION: Despite successful weight maintenance, the decrease in ambulatory blood pressure after rapid weight loss was largely transient. The increase in parasympathetic tone was more sustained, but also gradually attenuated during 1 year of weight maintenance. PMID- 12569269 TI - Atorvastatin causes depressor and sympatho-inhibitory effects with upregulation of nitric oxide synthases in stroke-prone spontaneously hypertensive rats. AB - OBJECTIVE: Recent studies have suggested that statins decrease blood pressure in hypertensive animals and upregulate endothelial nitric oxide synthase (eNOS) expression. However, the effects of statins on the expression of nitric oxide synthase (NOS) in the brain and the sympathetic nervous system remain to be elucidated. The aim of this study was thus to examine the effects of atorvastatin on blood pressure, sympathetic nerve activity, and the expression of NOS in stroke-prone spontaneously hypertensive rats (SHRSP) as well as in Wistar-Kyoto (WKY) rats. METHODS: The animals received atorvastatin (50 mg/kg per day) for 30 days. Systolic blood pressure and heart rate were evaluated using the tail-cuff method. Urinary norepinephrine excretion was measured for 24 h. The expression of eNOS, neuronal NOS (nNOS), and inducible NOS (iNOS) in the brain (cortex, cerebellum, hypothalamus and brainstem), aorta and heart were determined by Western blot analysis. RESULTS: Systolic blood pressure and 24-h urinary norepinephrine excretion were significantly decreased in SHRSP, but not in WKY, after the treatment with atorvastatin. The eNOS and iNOS expression in the brain and aorta was significantly increased in atorvastatin-treated SHRSP and WKY. However, the nNOS expression in the brain was not altered in the atorvastatin treated group. CONCLUSIONS: These results suggest that atorvastatin decreases blood pressure, at least in part via the reduction of sympathetic nervous system activity in SHRSP. They also suggest that this sympatho-inhibitory effect may be mediated by an increase in NO production, with the upregulation of eNOS expression in the brain. PMID- 12569270 TI - Acute effects of transdermal estrogen on hemodynamic and vascular reactivity in elderly postmenopausal healthy women. AB - OBJECTIVE: The acute effects of estrogen on hemodynamic responses were studied with emphasis on the sympathoadrenal system and peripheral circulation. DESIGN: Eleven healthy postmenopausal women recruited from the population-based study BEDA were included in this randomized, double-blind, cross-over, placebo controlled hypothesis-generating pilot study, where the effect of transdermal estrogen (17 beta-estradiol, 100 microg/24 h) was tested. METHODS: Twenty-four hours after the patch with estrogen/placebo was attached, the blood pressure during rest and mental stress test was measured, together with blood samples for analysis of P-adrenaline and P-noradrenaline. Twenty-four-hour ambulatory registration of blood pressure and heart rate were recorded. Contractile properties and endothelial function of subcutaneous small arteries from gluteal biopsies were studied with the wire-myograph technique. RESULTS: Estrogen treatment reduced both ambulatory systolic blood pressure (5 mmHg, P = 0.05), diastolic blood pressure (3 mmHg, P < 0.05) and heart rate (6-8 beats/min during morning hours, P < 0.01). Diastolic blood pressure during and after mental stress was significantly reduced after estrogen treatment (p < 0.01). The levels of P adrenaline and P-noradrenaline were similar in both treatment protocols. The contractile properties of the arteries were not significantly influenced by estrogen. Substance P induced nitric oxide-dependent relaxation in both estrogen treated and placebo-treated precontracted arteries. Acetylcholine, on the other hand, induced a non-nitric oxide, non-prostanoid-dependent hyperpolarization, which was inhibited by potassium-induced depolarization after placebo but not after estrogen treatment. CONCLUSIONS: Acute administration of transdermal estrogen in clinically relevant doses modulates hemodynamics, probably by an altered parasympathetic balance, which might involve changes at the muscarinic receptor level. PMID- 12569271 TI - Neutrophil activation and production of reactive oxygen species in pre-eclampsia. AB - OBJECTIVE: To investigate neutrophil NADPH oxidase activation and subsequent production of reactive oxygen species (ROS) in pre-eclampsia. DESIGN: Baseline values and the activated response of neutrophils upon stimulation of the NADPH oxidase with the agonists was measured. Neutrophils from 17 third-trimester pre eclamptic and 17 age- and gestation-matched normal pregnant women were examined. METHODS: Neutrophil ROS production was measured by both lucigenin- and luminol derived chemiluminescence. The abundance of the various phox proteins was examined using Western blotting techniques. Lucigenin-derived ROS generation was significantly increased in neutrophils isolated from women with pre-eclampsia compared with normotensive controls in the case of both agonists [n-formyl-met leu-phe (fMLP): pre-eclamptic 2.071 +/- 0.336 relative light units seconds (RLU.s) and normotensive 1.141 +/- 0.249 RLU.s, P = 0.035; phorbol-12-myristate 13-acetate (PMA): pre-eclamptic 34.954 +/- 2.634 RLU.s and normotensive 17.208 +/ 3.325 RLU.s, P = 0.0001]. Luminol-derived ROS generation was also significantly increased in the neutrophils isolated from the women with pre-eclampsia compared with the normotensive controls in the case of both agonists (fMLP: pre-eclamptic 1.955 +/- 0.316 RLU.s and normotensive 1.058 +/- 0.191 RLU.s, P = 0.023; PMA: pre eclamptic 4.108 +/- 0.351 RLU.s and normotensive 3.073 +/- 0.332 RLU.s, P = 0.042). There were no differences between the relative abundance of the phox proteins in the two groups. CONCLUSIONS: Neutrophils isolated from women with pre eclampsia during the third trimester showed increased sensitivity to agonist stimulation and produced significantly more ROS than age-matched normotensive controls. This was not due to an increased abundance of any of the phox proteins. Increased ROS production in pre-eclampsia may highlight a role for neutrophils in the oxidative stress and associated endothelial dysfunction that are characteristic of the condition. PMID- 12569272 TI - Increased salt sensitivity induced by impairment of sensory nerves: is nephropathy the cause? AB - OBJECTIVE: A unique model of hypertension developed in our laboratory shows that neonatal degeneration of capsaicin-sensitive sensory nerves renders a rat responsive to salt load with a significant rise in blood pressure. Considering that capsaicin impairs the micturition reflex, increased retention of urine in the bladder of capsaicin-treated rats may lead to nephropathy and therefore to an increase in blood pressure. The present study was designed to test the hypothesis that abnormalities in renal function and blood pressure in this model are independent of urine retention, and are reversible when a high-salt diet is withdrawn. DESIGN AND METHODS: Newborn Wistar rats were given either 50 mg/kg capsaicin subcutaneously or vehicle on the first and second days of life. After weaning, one set of male rats was treated for 3 weeks with: capsaicin pretreatment plus high-sodium diet (4%, CAP-HS), capsaicin plus normal-sodium diet (0.5%, CAP-NS), control plus high-sodium diet (CON-HS), and control plus normal sodium diet (CON-NS). The other set of male rats was given the same treatment for 4 weeks except that the two high-salt-treated groups were switched to normal salt (CAP-HS-NS and CON-HS-NS) for the last week. Chronic bladder catheterization was carried out in all rats to maintain free flow of urine. Western blot was used for measurement of the capsaicin (vanilloid) receptor levels in dorsal root ganglia. Radioimmunoassay was used for measurement of plasma renin activity and plasma aldosterone levels. RESULTS: Capsaicin (vanilloid) receptor contents in dorsal root ganglia were markedly decreased in all capsaicin-treated groups. Plasma renin activity and plasma aldosterone levels were higher in CAP-HS rats than in CON-HS rats, but were not statistically different between CAP-HS-NS and CON-HS-NS rats. Urine Na excretion but not urine volume was decreased and water intake increased in CAP-HS compared with CON-HS, but these parameters were not statistically different between CAP-HS-NS and CON HS-NS rats. Although systolic blood pressure and mean arterial pressure were higher in CAP-HS and CAP-HS-NS rats than in CON-HS and CON-HS-NS rats, they were higher in CAP-HS than in CAP-HS-NS rats. CONCLUSION: Despite elimination of urine retention, sensory denervation impairs renal function and leads to an increase in blood pressure in response to high salt intake. The abnormalities in renal function and blood pressure in sensory denervated rats are reversible, at least in part, when high salt intake is withdrawn. These data support the hypothesis that sensory innervation counterbalances the pro-hypertensive systems and serves as a modulator to control salt sensitivity and cardiovascular function. PMID- 12569273 TI - Reduced activity of the kallikrein-kinin system predominates over renin angiotensin system overactivity in all conditions of sodium balance in essential hypertensives and family-related hypertension. AB - OBJECTIVE: To study the renin-angiotensin-aldosterone and kallikrein-kinin systems in essential hypertensives and offspring of hypertensive parents during different sodium loads, and to explore their possible influence on renal hemodynamics. METHODS: Forty-five essential hypertensives (35 +/- 4 years old, 25 males), 30 offspring of hypertensive parents (26 +/- 8 years old, 16 males) and 30 normotensive controls (28 +/- 5 years old, 20 males) were submitted to three different sodium loads (high, 250 mmol/l; normal, 140 mmol/l; and low, 20 mmol/l). Blood pressure, plasma renin activity, serum aldosterone, total kallikrein and urinary kallikrein-like activity were measured after each period. Effective renal plasma flow and glomerular filtration rate were also measured. In essential hypertensive subjects, renal hemodynamic and hormonal parameters were also measured after 3 days of 20 mg enalapril administration. RESULTS: Plasma renin activity and serum aldosterone were higher in normotensives, essential hypertensives and offspring of hypertensive parents only during low sodium intake, whereas urinary kallikrein activity was lower in hypertensive offspring and essential hypertensives, compared with normotensives, during the three diet conditions. Effective renal plasma flow was found to be reduced in hypertensives and normotensive offspring, while the glomerular filtration rated was similar in the three groups. Angiotensin converting enzyme inhibitor (ACEI) administration to essential hypertensives for 3 days normalized effective renal plasma flow, increased plasma renin activity and decreased aldosterone and urinary kallikrein activity. CONCLUSIONS: Our observations confirmed the presence of a hormonal imbalance between the renin-angiotensin-aldosterone system and the kallikrein kinin system, not only in essential hypertensives but also in the offspring of hypertensive parents. This imbalance probably affects the renal circulation and sodium homeostasis, since there was reduced effective renal plasma flow in both populations compared with normotensive subjects. The positive effect of ACEI, resulting in normalization of the effective renal plasma flow in essential hypertensive patients, suggests the involvement of both systems in impaired renal circulation. PMID- 12569274 TI - The effect of ramipril on albumin excretion in diabetes and hypertension: the role of increased lysosomal activity and decreased transforming growth factor beta expression. AB - OBJECTIVES: Albumin excretion is modulated post-filtration by lysosomal processing that produces a spectrum of albumin-derived material in urine, much of which is not detected by conventional immunoassays. This study aimed to determine the efficacy of ramipril treatment (+ RAM) after 24 weeks on total albumin excretion (intact plus albumin-derived peptides) in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats with (d) and without (c) diabetes. METHODS: Intact albumin excretion was analysed by radioimmunoassay and total albumin excretion was analysed by measuring radioactivity derived from circulating [ C]albumin. Renal lysosomal activity was determined by urinary [ H]dextran sulphate desulphation. Renal transforming growth factor-beta 1 (TGF beta 1), TGF-beta inducible gene-h3 (beta ig-h3) and angiotensinogen mRNA production were analysed by real time reverse transcriptase-polymerase chain reaction. RESULTS: Hypertension (SHR-c and SHR-d) resulted in a significant increase in intact albumin excretion, which was significantly reduced by ramipril treatment (P < 0.05 for SHR-c + RAM and 0.001 for SHR-d + RAM compared to non treated). This was accompanied by a significant decrease in blood pressure (P < 0.001 for SHR-c + RAM and SHR-d + RAM), renal beta ig-h3 mRNA production (P < 0.05 for SHR-c + RAM and SHR-d + RAM), and an increase in lysosomal activity. Diabetes (WKY-d and SHR-d) primarily caused a significant increase in total albumin excretion, predominantly in the form of albumin-derived fragments in the WKY-d group and intact albumin in the SHR-d group. Ramipril treatment reduced total albumin excretion in the WKY-d + RAM group (P < 0.001). CONCLUSIONS: Ramipril prevents increases in both intact albumin and total albumin excretion in hypertensive and diabetic states, respectively. PMID- 12569275 TI - Ultrafast multiplanar determination of left ventricular hypertrophy in spontaneously hypertensive rats with single-shot spin-echo nuclear magnetic resonance imaging. AB - OBJECTIVE: To determine whether left ventricular hypertrophy can be correctly evaluated in hypertensive rats with a new nuclear magnetic resonance (NMR) imaging modality that is relatively simple to operate and provides results of constant quality while offering a high signal-to-noise ratio. DESIGN Left ventricular mass as calculated from the NMR imaging analysis was compared with the actual left ventricular mass measured by gravimetry. METHODS: Single-shot ultrafast spin-echo (SSFSE) imaging of hearts of Wistar-Kyoto rats and spontaneously hypertensive rats was performed at 4 T. Left ventricular mass was determined by using Simpson's rule on stacks of images acquired in systole and diastole. RESULTS: SSFSE NMR imaging performed in systole or in diastole evaluated and quantified left ventricular hypertrophy in hearts of spontaneously hypertensive rats very similarly to gravimetry. The left ventricular mass as determined by NMR was in good accordance with the actual left ventricular weight (SEE: 30.39 and 35.86 mg for the systolic and diastolic NMR acquisitions, respectively). CONCLUSION: Using an SSFSE sequence, high-quality NMR images of the rat heart can be generated very reliably with sufficient contrast and temporal and spatial resolution, and allow precise, non-invasive and fast characterization of left ventricular hypertrophy in a hypertensive rat model. PMID- 12569276 TI - Angiotensin II type 1 and endothelin type A receptor antagonists modulate the extracellular matrix regulatory system differently in diastolic heart failure. AB - OBJECTIVE: Ventricular fibrosis plays a pivotal role in the development of diastolic heart failure and is a therapeutic target; however, the effects of pharmacological interventions on the extracellular matrix (ECM) regulatory system in diastolic heart failure remain to be clarified. DESIGN AND METHODS: Dahl salt sensitive rats fed on a diet containing 8% NaCl from age 7 weeks--a hypertensive diastolic heart failure model--were divided into untreated rats, rats treated with angiotensin II type 1 (AT1) receptor antagonist and those treated with endothelin type A (ETA) receptor antagonist. RESULTS: Ventricular fibrosis progressed in the untreated rats, with increases in mRNA levels of type I collagen, matrix metalloproteinase-2, -9 and -13, and tissue inhibitor of metalloproteinase-1 and -2. Both antagonists attenuated ventricular fibrosis to the same degree. AT1 receptor blockade decreased the type I collagen mRNA level more than ETA receptor blockade. ETA receptor blockade did not decrease the matrix metalloproteinase-2 mRNA level that was decreased by AT1 receptor blockade, and decreased the tissue inhibitor of metalloproteinase-2 mRNA level that was not affected by AT1 receptor blockade. These led to a higher ratio of matrix metalloproteinase-2 to tissue inhibitor of metalloproteinase-2 mRNA levels and a greater 72-kDa gelatinase activity in the rats treated with ETA receptor antagonist than in those treated with AT1 receptor antagonist, and may well cancel out the lesser decline in collagen synthesis, resulting in the equivalent attenuation of ventricular fibrosis. CONCLUSIONS: AT1 receptor and ETA receptor antagonists provide their beneficial effects on the ECM through different modulation of its regulatory system. PMID- 12569277 TI - Ventricular repolarization is prolonged in nondipper hypertensive patients: role of left ventricular hypertrophy and autonomic dysfunction. AB - OBJECTIVE: The aim of this study was to investigate the influence of circadian behavior of blood pressure, left ventricular hypertrophy, and autonomic function on QTc interval duration in untreated hypertensive patients. DESIGN: Hypertensive patients underwent simultaneous blood pressure and ECG 24-h ambulatory monitoring. Patients were classified into two groups on the basis of a lack of nocturnal fall in blood pressure, as dippers and nondippers. A group of normotensive healthy subjects was studied as controls. METHODS: QT and QTc intervals were automatically computed and spectral analysis was applied to RR interval time series from the same electrocardiogram (ECG) recordings. Left ventricular mass index (LVMI) was computed by echocardiogram. RESULTS: No difference among the three groups was found concerning mean values and circadian pattern of heart rate; by contrast, QTc was significantly longer in nondippers compared to dippers or to normotensive subjects, particularly at night-time, whereas all groups exhibited similar circadian variations in heart rate. Compared to dippers, nondippers showed significantly higher LVMI, which positively correlated with QTc, and parasympathetic withdrawal, which negatively correlated with QTc. CONCLUSIONS: Nondippers show a prolonged ventricular repolarization throughout the 24-h period, absent either in dippers or normotensives. The association of left ventricular hypertrophy and vagal deactivation may lead to prolongation of QTc, potentially facilitating ventricular arrhythmias in nondipper hypertensive patients. PMID- 12569278 TI - Effects of several polyhydroxylated flavonoids on the growth of B16F10 melanoma and Melan-a melanocyte cell lines: influence of the sequential oxidation state of the flavonoid skeleton. AB - The response of B16F10 melanoma and Melan-a melanocyte cell lines to treatment with five polyhydroxylated flavonoids and gallic acid, after 24 and 72 h of exposure, was determined, and the relationship between any antiproliferative effects observed and the chemical structure is discussed. After 24 h, none of the studied compounds showed significant cytotoxic activity in the B16F10 cell line, whereas compounds with an adjacent trihydroxylated substitution pattern did affect the viability of the Melan-a cell line. After 72 h of exposure, myricetin, baicalein and gallic acid significantly inhibited both B16F10 and Melan-a cell cultures, whereas luteolin and quercetin had only a moderate effect. Eriodictyol only had an effect on Melan-a cell viability, which was reduced slightly. These results suggest that the presence of a C2-C3 double bond and three adjacent hydroxyl groups in the flavonoid A- or B-rings confers greater antiproliferative activity to the flavonoid. PMID- 12569279 TI - Enzyme and integrin expression by high and low metastatic melanoma cell lines. AB - Dissemination of a malignant tumour is the result of a cascade of events beginning with detachment of cells from primary tumour followed by extravasation and growth at secondary sites. The differences in metastatic ability could be attributed to properties intrinsic to the various tumour types. Thus the clonal selection of tumour cells from successive metastases apparently results in cells better equipped for survival and formation of colonies in secondary sites, indicating that survival is not a random phenomenon. Many studies of malignant cells have correlated the overexpression of adhesion receptors such as integrins and the production of cysteine proteases and glycosidases with the progression of malignancy. The interaction of cysteine proteases with basement membrane components has been implicated in tumour invasion, activation of hormones and growth factors. On the other hand, the expression of the heparanase gene and its protein has been associated with the metastatic potential of several human and mouse tumour cell lines. This study aimed to investigate the correlations between the metastatic properties of clones with high and low metastatic potential and their ability to adhere to the extracellular matrix and to degrade proteins and sulphated glycosaminoglycans present there. Clonal selection of the B16F10 cell line was performed, and the clones were examined for the expression of an integrin-type laminin receptor. A significantly higher level was detected in a high metastatic clone. Enzymatic assays showed higher activity for alpha-d-N acetylglucosaminidase, beta-d-N-acetylgalactosaminidase and beta-d-glucuronidase in conditioned medium from low metastatic clones compared with that from high metastatic clones. However, higher endopeptidase activity was observed in conditioned medium from high metastatic clones. In summary, these results showed a positive correlation between high metastatic potential and endopeptidase secretion. Similarly, a positive correlation was observed between low metastatic cells and the secretion of glycosaminoglycan-degrading glycosidases. PMID- 12569280 TI - Lipid peroxidation and antioxidant capacity in selected tissues of healthy black C57BL/6J mice and B16 melanoma-bearing mice. AB - During the process of melanogenesis free radicals are generated. The aim of this study was to investigate the effects of melanogenesis in B16 melanoma on lipid peroxidation and antioxidant capacity in selected tissues of black C57BL/6J mice. The study was conducted on 24 mice: 12 healthy controls and 12 with a transplanted B16 melanoma. Two weeks after the melanoma transplant, when the average weight of the tumours was approximately 2.0 g, blood samples were taken from the orbital venous plexus. The mice were killed by dislocation of the spinal cord, and the brain, liver and lungs were removed for analysis. The level of thiobarbituric acid-reactive substances (TBARS) and of 1,1-diphenyl-2 picrylhydrazyl (DPPH) reactive substances were determined in full liver, lung and brain homogenates and in serum. The activity of superoxide dismutase (SOD) was determined only in homogenized tissue. The concentration of TBARS and the SOD activity were statistically significantly higher in all the studied tissues from mice with B16 melanoma than in tissues from healthy mice. The antioxidant capacity, however, was lower in the tissues of melanoma-bearing mice. The results obtained demonstrate an increase in oxidative stress in the tissues of mice bearing a transplanted B16 melanoma. PMID- 12569282 TI - Microsatellite analysis at 1p36.3 in malignant melanoma of the skin: fine mapping in search of a possible tumour suppressor gene region. AB - Deletions in 1p36 in malignant melanoma have been found in high percentages in nodular melanomas and melanoma metastases. Despite many efforts, no candidate tumour suppressor gene associated with malignant melanoma has so far been found in this region. To further determine a possible tumour suppressor gene locus, we carried out a deletion mapping of chromosome 1p36 at nine microsatellite loci in 74 malignant melanomas. Loss of heterozygosity (LOH) in this region was found in 77% of nodular melanomas (NMs), 86% of metastatic melanomas, but only 20% of superficial spreading melanomas (SSMs). Regarding the allelic losses, the nodular and metastatic melanoma samples could be divided into three groups: one showing LOH at the more telomeric loci D1S243 and D1S468 (1p36.33), one displaying allelic loss at the more centromeric loci D1S214 and D1S253 (1p36.32-31) and one with LOH over all informative loci between D1S243 and D1S160. We did not find any significant correlation between a deletion in any of the investigated loci and the survival data of the patients. However, our results confine the deleted region in malignant melanoma to a very small area around 1p36.32, thus facilitating the search for the tumour suppressor gene with importance in malignant melanoma. PMID- 12569281 TI - Inverse relationship between density of cutaneous hair and pigmented lesion count in patients with malignant melanoma. AB - The aim of this study was to investigate whether there is an inverse relationship between the density of cutaneous hair and the number of pigmented lesions in patients with malignant melanoma, and to review the clinical and experimental findings with regard to this relationship. Cutaneous hair density and pigmented lesion counts were determined at nine sites by two physicians using specified criteria in 10 patients with a history of malignant melanoma and 22 control subjects. Study participants were Caucasian males of similar age, height and weight, and had similar occupational, ethnic, socioeconomic and sun exposure backgrounds. Statistical analysis was performed using combined data from all the sites and individual site data to determine whether an inverse relationship between cutaneous hair density and pigmented lesion counts exists. The results demonstrated statistically significant inverse relationships for the chest, upper back, upper arm and forearm in the melanoma patients, but not in the controls. For all the sites combined, both groups demonstrated statistically significant inverse relationships. The study findings are consistent with the hypothesis that some pigmented lesions may arise from the melanocytes of the hair follicle. This hypothesis may lead to new approaches to studying and treating melanoma. PMID- 12569283 TI - Sentinel lymph node biopsy in melanoma patients with clinically negative regional lymph nodes--one institution's experience. AB - The purpose of this prospective study of sentinel lymph node (SLN) biopsy in a large series of melanoma patients with clinically negative regional lymph nodes from one cancer centre was to analyse the reliability of the procedure, the pattern of failures during follow-up and the factors affecting the clinical outcome of patients. Between April 1995 and November 2001, 726 consecutive patients with primary cutaneous malignant melanoma underwent SLN biopsy with preoperative lymphoscintigraphy. The vital blue dye technique was used in 170 patients, and the blue dye technique combined with intraoperative lymphoscintigraphy in 556 patients. The primary melanoma sites were head and neck in nine patients, the extremities in 419 patients, and the trunk in 298 patients. The median Breslow thickness was 3.0 mm. All patients were followed closely, the median follow-up time being 34 months. The sentinel node(s) were successfully identified in 96% of patients. Intraoperative lymphoscintigraphy combined with the blue dye technique improved the SLN identification rate (technical success in 97.3% of cases) compared with the blue dye technique alone (technical success in 91.6%). The rate of failed SLN procedures was significantly (P = 0.007) lower in inguinal basins (3.1%) compared with axillary basins (7.9%). SLN metastases were detected in 147 patients (20.2%). The presence of SLN metastases correlated significantly with primary tumour thickness and ulceration (P < 0.001). The false negative SLN biopsy rate was 4.66% (27 out of 579 SLN-negative patients). All but two node-positive patients underwent complete lymphadenectomy. Lymph nodes other than SLNs were found to contain metastases in 26.9% of patients (39 out of 145). The 5 year overall survival (OS) rate was 84% for SLN-negative patients and 40% for SLN-positive patients. Five variables showed a strong, statistically significant negative independent prognostic association with OS: positive SLN status (P = 0.000001), primary melanoma thickness > 4 mm (P = 0.0009), male gender (P = 0.001), more than one lymph node involvement (P = 0.02) and lymph node extracapsular extension (P = 0.03). SLN biopsy is currently a valuable and effective diagnostic procedure for the precise staging of patients with clinically N0 cutaneous melanoma. So far SLN biopsy seems to be the only accessible method for consciously oriented detection of nodal micrometastases in melanoma that would otherwise go undetected. SLN status is the most important factor proven to distinguish high and low risk melanoma patients. PMID- 12569284 TI - Serum S100B is suitable for prediction and monitoring of response to chemoimmunotherapy in metastatic malignant melanoma. AB - Serum S100B and lactate dehydrogenase (LDH) levels were evaluated for their ability to predict response in patients with metastatic malignant melanoma and to determine their usefulness in monitoring the results of chemoimmunotherapy. Levels were studied in 53 patients with metastatic malignant melanoma receiving chemoimmunotherapy and in 19 control patients with metastatic renal cell carcinoma receiving a similar immunotherapy regimen. The serum S100B level was elevated in 81% of the patients before treatment. Marker levels were significantly higher in patients who did not respond (n = 22). Patients with S100B levels >or= 1.0 microg/l were less likely to obtain remission or stable disease than the group with normal or moderately elevated serum concentrations (P < 0.01). After treatment, 17 of the 31 (55%) patients with stable or responsive disease had a S100B serum level below the cut-off point versus only one of the 22 (5%) patients in the group with progressive disease. For LDH the proportions of patients were 17 out of 31 (55%) and nine out of 22 (41%), respectively. In 15 melanoma patients there was a transient rise in the level of serum S100B at the beginning of systemic therapy. All 19 patients in the control group had an initial serum S100B level or =65 years). All patients received at least 1 cycle of chemotherapy (median 3 cycles). Chemotherapy-associated toxicity was generally mild, more pronounced in females and did not increase with age. Median time to progression was 10.5+/-3.2 months for the patients < 65 years and 5.1+/-1 months for patients > or =65 years. median overall survival was 17.5+/-3.8 months in patients < 65 years and 8.6+/-1 months in patients > or =65 years (p < 0.0001). In multivariate analysis, age and female sex remained independent prognostic factors. Our data indicate that a treatment concept including concomitant radio- and chemotherapy is feasible even in elderly patients with GBM. PMID- 12569301 TI - Biweekly bolus 5-fluorouracil and leucovorin plus oxaliplatin in pretreated patients with advanced colorectal cancer: a dose-finding study. AB - The primary objective of this study was to determine the maximum tolerable dose (MTD) and dose-limiting toxicity (DLT) for bolus 5-fluorouracil (5-FU) administered on a biweekly schedule and in combination with fixed doses of leucovorin (LV) and oxaliplatin. The secondary objectives were to evaluate the toxicity profile and antitumor activity of this regimen for pre-treated patients with advanced colorectal cancer. A total of 26 patients with documented fluoropyrimidine-resistant, advanced colorectal cancer were enrolled into this phase I study. Fixed dose of oxaliplatin (85 mg/m2) was delivered as an i.v. infusion over 2 h, followed by LV (20 mg/m2) and 5-FU bolus every 2 weeks. The starting dose of 5-FU was 600 mg/m2, which was then incremented by 100 mg/m2 for each dose level. The DLT was determined for the first two treatment cycles, while toxicity and efficacy were evaluated throughout treatment. Six dose levels were tested. The MTD of 5-FU was deemed to be 1000 mg/m2 since dose-limiting fatigue was noted for three of the five-patient cohort during the first two cycles of chemotherapy at dose level 6. The most frequent treatment-related toxicities during the study were neutropenia, vomiting, diarrhea, fatigue and neuropathy. In an intent-to-treat analysis, the objective response rate was 30.8% (95% confidence interval 11.8-49.8%) for the 26 patients. The combination of bolus 5 FU/LV and oxaliplatin every 2 weeks is a feasible and effective treatment at the recommended dosages. A phase II study, to more-precisely define activity and toxicity, is ongoing. PMID- 12569302 TI - Thioredoxin reductase and cancer cell growth inhibition by organotellurium antioxidants. AB - Thioredoxin (Trx) expression is increased in several human primary cancers and the Trx/Trx reductase (TrxR) system therefore provides an attractive target for cancer drug development. Novel organotellurium antioxidants, especially a primitive analog of vitamin E (compound 1d) and compounds 7, 9 and 10--all carrying highly functionalized 4-(dialkylamino)phenyltelluro groups to secure high antioxidative capacity--were found to inhibit TrxR with IC50 values in the low micromolar range. Whereas antioxidant 1d also inhibited the growth of MCF-7 human breast cancer cells in culture at a similar level (IC50 = 1.8 microM), the other TrxR inhibitors were inactive in concentrations below about 10 M. PMID- 12569304 TI - Anti-cancer alkyl-lysophospholipids inhibit the phosphatidylinositol 3-kinase Akt/PKB survival pathway. AB - Synthetic alkyl-lysophospholipids (ALPs) represent a new class of anti-tumor agents that target cell membranes and induce apoptosis. However, the exact mechanisms by which ALPs exert these effects remain unclear. Here, we investigated in the epithelial carcinoma cell lines A431 and HeLa the effect of three clinically relevant ALPs [Et-18-OCH3 (Edelfosine), HePC (Miltefosine) and D 21266 (Perifosine)] on the phosphatidylinositol 3-kinase (PI3K)-Akt/PKB survival pathway. We found that growth factor-induced Akt/PKB activation in these cells is dependent on PI3K and that all three ALPs inhibited this pathway in a dose dependent manner. We further showed that inhibition of the PI3K-Akt/PKB pathway by wortmannin or ALPs is associated with activation of the pro-apoptotic SAPK/JNK pathway. Inhibition of the PI3K-Akt/PKB survival pathway represents a novel mode of action of ALPs that may significantly contribute to the induction of apoptosis. PMID- 12569303 TI - Preclinical in vitro evaluation of hematotoxicity of the cisplatin-procaine complex DPR. AB - We evaluated in vitro the inhibitory effect of cis-diaminechloro-[2 (diethylamino) ethyl 4-amino-benzoate, N4]-chlorideplatinum(II) monohydrochloride monohydrate (DPR) on colony formation by granulocyte/macrophage (CFU-GM) peripheral blood progenitor cells, representing a method to quantitate the toxicity of drugs to the hematopoietic system, and human leukemic cell lines. The results were compared with those obtained exposing cells to cisplatin and carboplatin. Our data showed that while DPR had a significantly better cytotoxic activity than cisplatin and carboplatin against HL60 and K562, and than carboplatin against Molt 4 cells, it showed 12 and 43 times less inhibitory effect on CFU-GM than cisplatin and carboplatin, respectively. These results suggest that the myelosuppressive activity of DPR could be lower than that of cisplatin and carboplatin, and, furthermore, that leukemic cells represent a preferential target for its cytotoxic activity compared to normal committed hemopoietic progenitor cells. All our results speak in favor of a better therapeutic index for DPR than for the other platinum compounds considered here. PMID- 12569305 TI - Differential effects of the optical isomers of KR30031 on cardiotoxicity and on multidrug resistance reversal activity. AB - The present study was performed to compare the cardiovascular adverse effects of verapamil, KR30031 and their optical isomers, and also to measure their ability to overcome multidrug resistance (MDR). The R-isomer of KR30031 (R-KR30031) was equipotent with the S-isomer of KR30031 (S-KR30031) and 25-fold less potent than the R-isomer of verapamil (R-verapamil) in relaxing the aorta isolated from rat (EC50: 11.8, 10.2 and 0.46 microM, respectively). The effect of R-KR30031 in decreasing left ventricular pressure of heart isolated from rat was 2- and 267 fold smaller than those of S-KR30031 and R-verapamil, respectively (EC50: 23.9, 9.4 and 0.089 mM, respectively). The hypotensive effect of R-KR30031 in rat was about 2- and 23-fold smaller than those of S-KR30031 and R-verapamil, respectively (ED20: 1.15, 0.60 and 0.05 mg/kg, respectively). On the other hand, R-KR30031 elicited potency similar to those of S-KR30031 and R-verapamil in enhancing the paclitaxel-induced cytotoxicity to HCT15/CL02 and MES-SA/DX5 cells that reveal high levels of P-glycoprotein expression (IC50: 3.11, 3.04 and 2.58 microM, respectively). In addition, the intrinsic cytotoxicity of R-KR30031 in HCT15/CL02 and MES-SA/DX5 cells was observed only at the very high concentration of 100 microM. All these results suggest that R- and racemic KR30031 are active modulators of MDR with potentially minimal cardiovascular adverse activity. PMID- 12569306 TI - Magnetic Resonance in breast imaging. PMID- 12569307 TI - Breast MRI: report on a multicentric national trial by the Study Section of Magnetic Resonance and Breast Imaging. PMID- 12569308 TI - Bronchial carcinoid tumors: clinical and radiological correlation. AB - PURPOSE: To evaluate the incidence of bronchial carcinoid tumor (BCT) in population affected by various lung tumors, retrospectively reviewed between 1986 2001, and to emphasize the radiographic patterns if they are characteristic. MATERIALS AND METHODS: In the above mentioned period, chest x-ray and CT were performed in 1110 patients, both male and female, affected by lung neoplasms. RESULTS: 20 patients were affected by BCT (16 males, and 13 females), aged between 26 and 75 years (mean age 57.5), with histopathologic diagnosis of typical (9 cases, 31%) and atypical (20 cases, 69%) bronchial carcinoid tumors. Localized right lung lesion were as follows: 7 cases in superior, 9 in middle and 4 in inferior lobes; in the left lung, 5 cases in superior, and 4 in inferior lobes. Size of the lesions was as average 2.4 cm in diameter (with a range of 0.5 12 cm). On x-ray and CT images, BCT appeared as a well marginated nodule, of which 9 were peripheral and 20 central. DISCUSSION AND CONCLUSION: BCT are classified as neuroendocrine carcinomas,and are divided in typical and atypical forms, with variable grade of malignancy. Central neoplasms are symptomatic due to bronchial obstruction (i.e., pneumonia, atelectasis, bronchiectasis, emphysema and/or lung abscess); if airway obstruction is partial, then cough, wheezing and recurrent pulmonary infections occur. Peripheral tumors are generally asymptomatic and they are discovered occasionally, when chest x-ray is made for other reasons. Radiographic features are similar in typical and atypical BCT. In central tumors a rounded well circumscribed hilar mass is noted, with lobulated or bumpy margins. Central cavitation is not referred to. Peripheral BCT appear as a solitary nodule, inferioer then 3 cm in size, marginated, surrounded by normal pulmonary tissue. Signs and symptoms of BCT are evasive and vague. No current clinical or laboratory procedures are useful in confirming the diagnosis; particularly, no imaging modalities are able to differentiate between BCT and other pulmonary tumors. For this reason, a clinical radiologic endoscopic and histopathologic approach is necessary. CT is more sensiticve then conventional radiography, especially in detecting small lesions, calcification and enlarged lymph nodes. MRI may be useful in those patients, who cannot tolerate IV contrast media. Scintigraphy may be employed in discovering relapses and long standing metastases. PMID- 12569309 TI - Characterization of focal hepatic lesions in cirrhotic patients by Pulse Inversion Harmonic Imaging US contrast specific technique with Levovist. AB - PURPOSE: To evaluate the capabilities of Pulse Inversion Harmonic Imaging (PIHI) with hepatospecific US contrast agent Levovist in the characterization of focal liver lesions in cirrhotic patients. MATERIALS AND METHODS: Thirty-nine focal hepatic lesions in 25 consecutive cirrhotic patients identified by conventional ultrasound (US), were evaluated by color Doppler (CD), power Doppler (PD) with spectral analysis of tumoural vessels and PIHI. PIHI was performed 30 seconds (vascular phase) and 3-5 minutes (late phase) after Levovist injection. To definitely characterize the evaluated focal hepatic lesions, helical-CT (HCT) enhancement patterns (15 patients) and/or surgical/bioptic histologic findings (10 patients) were considered as reference procedures. RESULTS: Thirty focal hepatic lesions classified as hepatocellular carcinoma (HCC) by reference procedures appeared hypoechoic (n=19), isoechoic (n=5) or hyperechoic (n=6) on conventional US, with basket arterial pattern (n=10), vessels within the tumor (n=6), peripheral arterial pattern (n=4) or no vascular pattern (n=10) on CD/PD evaluation. On PIHI they appeared hyperechoic (n=26) or isoechoic (n=4) in the vascular phase, if compared to the surrounding liver parenchyma, and hypoechoic (n=23) or isoechoic (n=7) in the late phase. Four focal hepatic lesions classified as regenerative nodules (RNs) by reference procedures appeared hypoechoic on conventional US, with peripheral venous/arterial pattern (n=1) or no vascular pattern (n=3) on CD/PD. On PIHI they appeared hypoechoic (n=3) or isoechoic (n=1) in the vascular phase, remaining prevalently hypoechoic (n=3) or isoechoic (n=1) in the late phase. Five focal hepatic lesions classified as hemangioma by reference procedures appeared hyperechoic (n=4) or hypoechoic (n=1) on conventional US with few peripheral venous vessels on CD/PD. On PIHI they revealed progressive fill-in from the periphery toward the centre during the vascular and late phase after Levovist injection. CONCLUSIONS: PIHI seems to be a reliable technique to characterize focal lesions in cirrhotic patients. PMID- 12569310 TI - Spiral hydro-CT in the evaluation of colo-sigmoideal cancer. AB - PURPOSE: To evaluate the accuracy of spiral hydro-CT in the diagnosis and preoperative staging of colo-sigmoideal neoplasms and the impact of this technique on therapeutic decisions. MATERIALS AND METHODS: Sixty patients with colo-sigmoideal neoplasms, 16 in caecum-ascending colon, 2 in transverse colon, 14 in descending colon, and 28 in sigmoid colon, diagnosed with fiberoptic colonoscopy and histopathological specimen, under-went spiral hydro-CT with IV contrast medium administration. Local invasion (T factor) and nodal involvement (N factor) were evaluated. In all cases, the CT reports were compared with the result of the histopathological examination of the surgical specimen. TNM and Dukes staging classifications were applied. RESULTS: Local invasion was correctly identified in 80% of cases. There was overstaging in 18.3% and understaging in 1.7% of the patients. Sensitivity, specificity and accuracy for nodal involvement were 57.9%, 67.5% and 63.3%, respectively. There was overstaging in 23.3% and understaging in 15% of patients. With Dukes classification, CT correctly staged 38.8% of patients. Significant differences were detected in different stages. The stage with the highest concordance was A/B1 with 58.8% (10/17), whereas the one with the lowest concordance was B2 with 9.1% (2/22). All of the three patients in stage D were correctly staged and none of the patients with stage C was overstaged. CONCLUSIONS: Spiral hydro-CT with intravenous contrast medium administration, despite the limits in the correct evaluation of local depth invasion and nodal involvement, proved useful for selecting patients likely to benefit from surgery. PMID- 12569311 TI - The usefulness of spiral Computed Tomography and colour-Doppler ultrasonography to predict portal-mesenteric trunk involvement in pancreatic cancer. AB - PURPOSE: The aim of this study was to determine the clinical utility of spiral computed tomography (CT) and colour Doppler ultrasonography (US) in the evaluation of portal-mesenteric trunk (PMT) involvement in pancreatic cancer. MATERIALS AND METHODS: Ninety-five patients with pancreatic cancer underwent preoperative assessment of the PMT with spiral CT and colour Doppler US. Five stages of vascular involvement were established. During surgery intraoperative US was performed to confirm the preoperative findings. RESULTS: Of the 95 patients observed, 82 (86.3%) underwent surgery. The sensitivity of spiral CT was 98%, specificity 79%, overall accuracy 80.2%. The positive predictive value was 87.5%; the negative predictive value 96%. The results of colour Doppler US were 92.3%, 72,7%, 72.8%, 79.5% and 88.8%, respectively. CONCLUSIONS: The results indicate that spiral CT is the gold standard in detecting PMT involvement in pancreatic cancer. Colour Doppler US is useful, but adds nothing to CT. Both of these techniques improve the possibility of predicting the resectability of pancreatic cancer. PMID- 12569312 TI - Hemoperitoneum following ovarian cyst rupture: CT usefulness in the diagnosis. AB - PURPOSE: The rupture of an ovarian cyst is among the most frequent causes of hemoperitoneum in women, and especially in young women. An ultrasound (US) examination performed in emergency allows the easy detection of fluids leaking into the abdomen. It may however be difficult to establish whether this fluid is blood and identify the cyst or the signs of a cystic rupture. The aim of the present study was the assessment of CT diagnostic capabilities in cases of hemoperitoneum following cyst rupture. MATERIALS AND METHODS: CT abdominal studies performed in emergency in 15 patients with surgical findings of ovarian cyst rupture and consequent hemoperitoneum were retro-spectively reviewed. An ultrasound examination carried out in 12 women was positive in all cases for abdominal effusion, without however confirming the presence of an ovarian cyst or without definitely defining the cystic rupture. Spiral CT examinations were performed with a preliminary unenhanced study of the abdomen (10-mm thickness image acquisitions, 10-mm intervals) and with scans obtained after injection of 150 cc of contrast medium (c.m.) at the speed of 2 cc/sec, 60-sec delay; late phase acquisition scans of the pelvic cavity were obtained in eight of the 15 patients. RESULTS: CT imaging allowed the depiction of the effusion in all cases. This was limited to the pelvic cavity in five cases and extended to the upper abdomen in the remaining ten patients; in seven of these ten cases, a difference in the density between the upper abdomen and the pelvic cavity was detected. An ovarian cyst was visualized in 11 patients; in eight of them, an irregular opacification of the walls was recorded, suggesting the diagnosis of cystic rupture. In the remaining four cases, a c.m. extravasation was observed, starting from the adnexal region, intracystic in one of them. In all these patients, a hyperdense collection in the pelvic cavity was documented in late-phase acquisitions. DISCUSSION AND CONCLUSIONS: CT imaging in cases of bleeding due to ovarian cyst rupture permits the acquisition of useful diagnostic information: a different effusion density in the pelvic cavity and in the upper abdomen, the direct visualization of the cystic mass (even when involved in a large effusion), the irregularity of opacified cystic walls (a sign of cyst rupture), and finally, the direct extravasion and collection of the c.m. in the pelvic cavity. These is fundamental information in those cases in whom US findings are negative or doubtful. PMID- 12569313 TI - Percutaneous treatment of femoro-popliteal aneurysms with covered stents. AB - PURPOSE: To assess the efficacy of the percutaneous treatment of femoro-popliteal aneurysms with covered stents. MATERIALS AND METHODS: Ten femoro-popliteal (1 femoral, 6 femoro-popliteal, 3 popliteal) aneurysms with diameters ranging from 2.1 to 6 cm (mean 3.6 cm) and lengths of 2.2-9 cm (mean 5.8 cm) were treated between September 1998 and December 2001. The patients were 8 men and 2 women aged between 33 and 73 years (mean age 65.4). Self-expanding covered stents (Wallgraft) were employed in all cases: the flexible and elastic stents were implanted percutaneously, under local anesthesia, using 9-10 F introducer sheaths. Color-Doppler ultrasound confirmed the clinical diagnosis. In all cases spiral CT angiography was performed to better evaluate the diameters and lengths of the aneurysms and the size of the arteries above and below the aneurysms. The patients were followed up with color-Doppler US and only occasionally with CT angiography. RESULTS: Stent implantation was successful in all patients; there were no complications. As regards follow up, 6 patients did not show any stent graft occlusions. One patient, who died 45 days later of causes unrelated to the procedure, was not taken into account. As for the remaining five patients, color Doppler US showed primary patency of the stents and exclusion of aneurysms at 24 months (2/5), 12 months (1/5), and 6 months (2/5). There were 4 cases of stent graft occlusion within the first month after placement (in 3/4 cases the caudal end of the aneurysm was located in the median segment of the popliteal artery), re-canalized by local thrombolysis and, in two cases, by PTA aimed at removing the stenoses caused by intimal hyperplasia: one case was patent at 36 months, 3 cases re-occluded and were converted to surgical by-pass (1 at 14 months, 2 at 18 months). Therefore in our series (mean follow-up: 18 months) primary patency was 55.5% and secondary patency was 66.6%. CONCLUSIONS: Percutaneous placement of a covered stent seems to provide an alternative to surgery for aneurysms with limited length and caudal end located not beyond the proximal tract of the popliteal artery. A good peripheral run-off (at least two patent leg arteries) is also necessary. Nevertheless, further studies and longer follow-up are required to confirm the results obtained in our small series. PMID- 12569314 TI - Acute hypoxic encephalopathy in the full-term newborn: correlation between Magnetic Resonance Spectroscopy and neurological evaluation at short and long term. AB - PURPOSE: To determine which brain metabolite ratios have the strongest correlation with poor neurological outcome in newborns with perinatal asphyxia, whether the correlation is stronger with basal ganglia (BG) and whether a combined approach using Magnetic Resonance Imaging (MRI) and Magnetic Resonance Spectroscopy (MRS) can be used to evaluate the severity of neonatal asphyxia. MATERIALS AND METHODS: Twenty newborns with perinatal asphyxia were studied within the first week from the hypoxic insult with MRI and MRS. The MRS variables were compared with the assessment of general movements (GMs). The brain metabolite levels measured by MRS were N-acetylaspartate (NAA), creatine (Cr), choline (Cho) and lactate and the ratios NAA to Cr, Lac to Cr, Cho to Cr, mI to Cr. RESULTS: High lactate levels and low NAA levels were found in the newborns with the worst outcome; the levels in newborns with good outcome were within normal limits. CONCLUSIONS: Correlations between NAA/Cr and Lac/Cr ratios, general movements and outcome at 6 months are stronger in the basal ganglia regions than in the frontal border zone. The presence or absence of lactate may indicate the severity of the brain insult and the combination of MRS, MRI and evaluation of general movements may assist in localising and predicting a long term brain injury. PMID- 12569315 TI - A comparison between two different concentration of contrast media with multidetector CT for the study of abdominal vascular system. AB - PURPOSE: To evaluate two different concentrations of contrast medium (CM) and two different acquisition techniques to obtain vascular images with multislice spiral CT (msCT). MATERIALS AND METHODS: Thirty patients referred for msCT examinations of the kidney or upper abdomen underwent evaluation of the abdominal aorta during the arterial phase, after giving their consent (GE Light Speed Plus). In the first part of the study (20 patients) the enhancement values measured at six different levels of the aortoiliac axis (celiac trunk, suprarenal aorta, subrenal aorta, aortal bifurcation, right iliac and left iliac) were compared after administering two different concentrations of iodinated contrast medium (300 and 370 mgI/ml) with two different acquisition techniques (HQ and HS). The flow rate was 3 ml/sec. In the second part of the study 10 patients were examined by changing the flow rate (2 and 4 ml/sec). Both quantity and quality assessments were conducted on axial images and on vascular reconstructions. RESULTS: In the first part of the study the best results were obtained with the 370 mgI/ml concentration and with the HQ acquisition technique (pitch 3, table feed 7.5 mm/rotation, gantry rotation speed 0.5 s). The quality of reconstructions gradually improved before reaching 240 HU, above which image quality remained constant. The patients examined with a flow rate of 2 ml/sec showed a poor image quality, unlike those examined with a flow rate of 4 ml/sec where we observed the maximum threshold values and constant optimal vascular opacification, with no significant differences from the patients studied at 3 ml/sec. CONCLUSIONS: Our results indicate that, albeit on a limited number of patients, a higher concentration of CM and the use of an HQ technique allow a high quality examination to be obtained. The HS technique yields results comparable to those obtained with higher concentrations. PMID- 12569316 TI - Emergency departments. PMID- 12569317 TI - OS acromiale producing rotator cuff impingement and rupture. A case report. PMID- 12569318 TI - Inflammatory pseudotumor of the liver: about a case investigated by US and CT. PMID- 12569319 TI - CT diagnosis of pancreatic lipoma: a case report and Literature review. PMID- 12569320 TI - Trans-femoral retrieval of a pace-maker lead from a thrombosed left succlavian vein. PMID- 12569321 TI - Percutaneous embolization of a gastroduodenal artery aneurysm secondary to antrectomy and Roux en Y reconstruction. PMID- 12569322 TI - Gigantiform cementoma of the jaw. A case report. PMID- 12569325 TI - Prognostic significance of ischemic electrocardiographic changes during vasodilator stress testing in patients with normal SPECT images. AB - BACKGROUND: Patients with ischemic electrocardiographic (ECG) findings during exercise stress testing but normal perfusion images generally have a low risk of cardiac death or myocardial infarction (<1% per year). During vasodilator stress testing, however, the prognostic significance of the combination of normal perfusion images and ischemic ECG changes is unknown. METHODS AND RESULTS: Among 5526 patients who underwent vasodilator stress single photon emission computed tomography (SPECT), 49 (0.9%) had normal images but ischemic ECG changes. A unique feature of this population was that 43 (88%) were women with a mean age of 67 +/- 10 years. Ischemic ECG changes occurred at a mean heart rate of 101 +/- 15 beats per minute and persisted for 6.8 +/- 4.7 minutes after termination of drug infusion. During follow-up of 28 +/- 20 months, cardiac death occurred in 2 patients and nonfatal myocardial infarction in 4 patients. The rate of cardiac death or nonfatal myocardial infarction was 4% at 1 year, 10% at 2 years, and 14% at 3 years. Of the 12 patients who underwent coronary angiography or autopsy during follow-up, 11 had multivessel coronary artery disease, indicating that these patients likely had false-negative SPECT image results. Eight patients required coronary revascularization. CONCLUSIONS: The finding of ischemic ECG changes with normal SPECT images during vasodilator infusion is uncommon, occurs primarily in older women, and is associated with a higher subsequent cardiac event rate than is customarily associated with normal images. PMID- 12569326 TI - Prognostic significance of ischemic electrocardiographic changes during adenosine infusion in patients with normal myocardial perfusion imaging. AB - BACKGROUND: The development of ST-segment depression during adenosine myocardial perfusion imaging (MPI) has been shown to be an independent predictor of subsequent cardiac events and worse outcome, particularly in association with ischemic defects. However, the prognostic significance of ST-segment depression with adenosine in patients with normal MPI is not known. METHODS AND RESULTS: We performed a retrospective analysis of 3231 patients undergoing adenosine MPI. Patients with baseline electrocardiographic (ECG) abnormalities were excluded. Overall, 228 patients (7%) had ischemic ECG changes develop during adenosine infusion. Of these, 66 (29% [2% of all patients]) had normal MPI (+ECG group). An age- and sex-matched group of 200 patients with normal MPI without ECG changes served as control subjects (-ECG group). During a mean follow-up of 29 +/- 12 months, patients in the +ECG group had significantly more adverse cardiac events than those in the -ECG group (nonfatal myocardial infarction, 7.6% vs 0.5%, respectively, P = .004; subsequent revascularization, 13.6% vs 2.5%, respectively, P = .0015). Although cardiac death alone did not differ between the 2 groups (+ECG, 3.0%, vs -ECG, 1.0%; P = .25), cumulative survival free from cardiac death and nonfatal myocardial infarction was worse in patients with ST segment depression during adenosine infusion and normal MPI (+ECG, 10.6%, vs ECG, 1.5%; log-rank chi(2) = 11.82; P = .0006). CONCLUSIONS: Patients with normal myocardial perfusion images in whom ST-segment depression develops during adenosine administration appear to be at increased risk for future cardiac events compared with similar patients without ECG evidence of ischemia. Ischemic ECG changes during adenosine MPI should warrant further evaluation, even when perfusion images are reassuring. PMID- 12569328 TI - Simultaneous cardiac output and regional myocardial perfusion determination with PET and nitrogen 13 ammonia. AB - BACKGROUND: The purpose of this study was to evaluate the possibility of measuring cardiac output during positron emission tomography (PET) examination of myocardial perfusion with nitrogen 13 ammonia. METHODS AND RESULTS: In 7 patients undergoing right-sided cardiac catheterization for evaluation of heart failure and 6 patients who had undergone heart transplantation, a thermodilution catheter for continuous measurement of cardiac output was inserted. An N-13 ammonia scan of the heart was subsequently performed, and with use of factor analysis, the time-activity curve from the right ventricle was derived from the dynamic image sequence. The PET-derived cardiac output was subsequently obtained according to the Stewart-Hamilton principle as the amount of injected tracer divided by the area under the time-activity curve. PET-acquired cardiac output measurements correlated closely with the invasively determined values for a wide range of cardiac output values (P < .0001). The mean difference was 0.12 L/min, with an SD of 0.74 L/min. The interobserver variation was low, with a mean difference of 0.06 L/min and an SD of 0.46 L/min. CONCLUSIONS: Cardiac output determination with N-13 ammonia and PET appears to be both accurate and precise and can be performed simultaneously with measurement of myocardial perfusion. PMID- 12569327 TI - Relationship between contractile reserve, Tl-201 uptake, and collateral angiographic circulation in collateral-dependent myocardium: implications regarding the evaluation of myocardial viability. AB - BACKGROUND: The aim of this study was to compare collateral angiographic circulation (CC), thallium 201 single photon emission computed tomography (SPECT), low-dose dobutamine echocardiography (LDDE), and their combination in identifying viable myocardium beyond a chronic occlusion. METHODS AND RESULTS: We studied 33 consecutive patients with a chronic occluded coronary artery and regional ventricular dysfunction who underwent dobutamine infusion (5-10 microg x kg(-1) x min(-1)), Tl-201 SPECT, and coronary revascularization (15 patients underwent coronary artery bypass grafting and 18 patients percutaneous transluminal coronary angioplasty). Echocardiography and Tl-201 SPECT at rest were repeated 90 +/- 48 days after revascularization. For viability assessment, Tl-201 SPECT showed the best sensitivity and accuracy (77% and 72%, respectively) compared with LDDE (64% and 63%, respectively) and CC (74% and 55%, P < .05 vs Tl 201 SPECT). Specificity was significantly better for both Tl-201 SPECT (65%) and LDDE (62%) compared with collateral circulation (27%, P < .001 vs Tl-201 SPECT and LDDE). Furthermore, combined Tl-201 SPECT and LDDE data improved specificity (from 88% to 92%) and positive predictive power (from 75% to 78%) but not global accuracy. A direct relationship between the number of viable segments and global functional recovery after revascularization was found only in the case of Tl-201 SPECT (r = 0.48, P = .04). CONCLUSIONS: This study demonstrates that CC has a very low specificity in the evaluation of viability in collateral-dependent myocardium and that LDDE has a very low sensitivity, whereas Tl-201 SPECT is the most reliable method, particularly in akinetic segments, and optimally identifies patients who will mostly benefit from revascularization in terms of ventricular functional improvement. No additional advantages in terms of accuracy were found when Tl-201 SPECT and LDDE data were combined. PMID- 12569329 TI - Assessment of myocardial viability in dysfunctional myocardium by resting myocardial blood flow determined with oxygen 15 water PET. AB - BACKGROUND: There is controversy about the role of decreased resting blood flow as the pathophysiologic correlate of hibernating myocardium. The aim of this study was an absolute quantification of volumetric myocardial blood flow (MBFvol) in dysfunctional myocardium with different viability conditions as defined by fluorine 18 deoxyglucose (FDG) positron emission tomography (PET) while taking into consideration the functional recovery after revascularization. The impact of MBFvol in the diagnosis of functional recovery was also investigated. METHODS AND RESULTS: Forty-two patients with severe coronary artery disease and dysfunctional myocardium underwent resting oxygen 15 water PET, as well as FDG PET and technetium 99m tetrofosmin single photon emission computed tomography, all attenuation-corrected. Relative FDG and Tc-99m tetrofosmin uptake (normalized to the segment with 100% Tc-99m tetrofosmin uptake), as well as MBFvol (myocardial blood flow multiplied by the water-perfusable tissue fraction to account for the flow to the entire segment volume), were determined in 18 myocardial segments per patient. Viability in dysfunctional segments (estimated by ventriculography) with reduced Tc-99m tetrofosmin uptake of 70% or lower was classified as viable (FDG >70%, mismatch) or nonviable (FDG < or =70%, match). Fifteen patients underwent revascularization and were followed up. Mismatch segments with improved function were classified as hibernating myocardium. Mean MBFvol in viable myocardium was slightly reduced (0.60 +/- 0.02 mL x min(-1) x mL(-1)) compared with that in normokinetic myocardium (0.64 +/- 0.01 mL x min(-1) x mL(-1)) (P = .036) and was significantly higher than in nonviable myocardium (0.36 +/- 0.01 mL x min(-1) x mL(-1)) (P < .001). Receiver operating characteristic analysis confirmed an FDG uptake greater than 70% as the optimal threshold to predict functional recovery (diagnostic accuracy [ACC], 76%). MBFvol in hibernating myocardium (0.62 +/- 0.04 mL x min(-1) x mL(-1)) was not significantly reduced compared with that in normokinetic myocardium (0.66 +/- 0.02 mL x min(-1) x mL(-1)) and was significantly higher than in persistently dysfunctional myocardium (0.51 +/- 0.04 mL x min(-1) x mL(-1)) (P < .05). The ACC of MBFvol greater than 0.40 mL x min( 1) x mL(-1) as the threshold to predict functional recovery was 61% but did not improve the accuracy of FDG PET by itself. CONCLUSIONS: In patients with severe coronary artery disease and dysfunctional myocardium, MBFvol as determined with O 15 water differs significantly between viable and nonviable myocardium as determined by FDG PET and is not significantly reduced in hibernating compared with normokinetic myocardium. Therefore chronically reduced resting blood flow appears unlikely to be the pathophysiologic correlate of the functional state of hibernation. However, MBFvol does not improve the ACC of FDG PET by itself. PMID- 12569330 TI - Differential systolic and diastolic effects of beta-adrenergic stimulation in patients with severe left ventricular dysfunction: a radionuclide ventriculographic study. AB - BACKGROUND: An attenuated inotropic response to beta-adrenergic stimulation has been documented in patients with severely depressed left ventricular (LV) function. Scant data exist regarding the effect of beta-adrenergic stimulation on LV diastolic function in human beings. Our objective was to evaluate the effect of dobutamine infusion on LV systolic and diastolic function in patients with severe ventricular dysfunction. METHODS AND RESULTS: We studied 26 patients (60 +/- 9 years) in stable condition with healed myocardial infarction and LV ejection fraction (EF) less than 35% by radionuclide ventriculography. LVEF, volumes, peak ejection rate (PER), peak filling rate (PFR), and peak systolic pressure-to-end-systolic volume ratio were calculated at the infusion rate of 5 and 10 gamma x kg(-1) x min(-1). At 5 gamma x kg(-1) x min(-1), no changes were observed in heart rate (75 +/- 16 beats/min vs 75 +/- 15 beats/min at rest), LVEF (23% +/- 8% vs 22% +/- 7%), and PER (1.40 +/- 0.48 end-diastolic volume per second [EDV/s] vs 1.30 +/- 0.48 EDV/s); however, a significant increase in PFR was observed (1.11 +/- 0.36 EDV/s vs 0.86 +/- 0.30 EDV/s, P < .01). At 10 gamma x kg(-1) x min(-1), an increase in LVEF (25% +/- 10%, P < .01) and PER (1.60 +/- 0.57 EDV/s, P < .01) was observed; PFR (1.25 +/- 0.36 EDV/s, P < .05) also increased, whereas heart rate (78 +/- 18 beats/min) was unchanged. No significant changes in LV volumes or in indices of LV contraction synchronicity were observed during infusion. Finally, the peak systolic pressure-to-end-systolic volume ratio was unchanged at 5 gamma x kg(-1) x min(-1) and significantly increased at 10 gamma x kg(-1) x min(-1). CONCLUSIONS: In stable patients with ischemic heart disease and severe LV dysfunction, beta-adrenergic stimulation may still induce a positive lusitropic response, despite attenuated inotropic and chronotropic responses; moreover, the effects on diastolic function occur earlier than those on systolic function. PMID- 12569331 TI - Use of atropine in patients with submaximal heart rate during exercise myocardial perfusion SPECT. AB - BACKGROUND: Failure to reach 85% of maximal predicted heart rate (MPHR) during exercise may render a myocardial perfusion single photon emission computed tomography (MPS) study nondiagnostic for ischemia detection. Although commonly used to increase heart rate (HR) during dobutamine stress, the administration of atropine for patients failing to achieve 85% of MPHR during exercise performed for MPS is still infrequent. METHODS AND RESULTS: Patients undergoing dual isotope MPS were considered candidates for the study when, during exercise treadmill testing, they had less than 85% of MPHR and were unable to continue because of fatigue, without an ischemic response. Forty-seven patients (aged 65.3 +/- 12.5 years, 78.7% men) received atropine (0.6-1.2 mg). Maximal HR achieved before and after atropine was 118.0 +/- 14.8 beats/min (76.3% +/- 6.2% of MPHR) and 146.4 +/- 12.6 beats/min (94.4% +/- 8.1% of MPHR), respectively (P < .001). Of patients, 44 (93.6%) reached at least 85% of MPHR after atropine and had diagnostic MPS studies. After atropine, arrhythmias occurred in 14 patients (29.8%) and other minor side effects in 1 (2.1%). CONCLUSIONS: Atropine allows patients initially failing to achieve 85% of MPHR during exercise to increase HR and have a diagnostic MPS study, without major complications. It may provide an alternative to pharmacologic stress for patients with a blunted HR response to exercise. PMID- 12569332 TI - Detection and reproducibility of mental stress-induced myocardial ischemia with Tc-99m sestamibi SPECT in normal and coronary artery disease populations. AB - BACKGROUND: Mental stress-induced ischemia, as detected by radionuclide studies, has yielded reversible ischemia in only 30% to 60% of patients with exercise induced ischemia. Little is known about the reproducibility of myocardial perfusion imaging in detecting mental stress-induced ischemia. The purpose of this study was to further evaluate the occurrence and reproducibility of mental stress-induced ischemia in patients with coronary artery disease (CAD) and in normal control subjects with a low likelihood of CAD by using sestamibi single photon emission computed tomography (SPECT) imaging. METHODS AND RESULTS: A total of 40 patients were enrolled in this study: 19 patients with CAD and typical angina or reversible ischemia (positive exercise treadmill study or positive adenosine thallium study) and 21 normal control subjects underwent mental stress testing as well as myocardial perfusion imaging. The subjects were given a speaking task, and SPECT imaging was subsequently performed. Two experienced readers compared mental stress imaging with a resting image using a 20-segment cardiac model. Hemodynamic changes in blood pressure and heart rate with mental stress were also measured in all subjects. Each patient with CAD also underwent repeat mental stress testing and myocardial imaging approximately 2 weeks later. Of the 19 patients with CAD and typical angina or with evidence of reversible ischemia, 16 (84%) demonstrated ischemia with mental stress, as detected by sestamibi SPECT imaging. The mean number of new or worsened perfusion defects attributable to mental stress was 3.5, with a mean severity of 1.7. These results were also reproducible. With repeated mental stress testing and myocardial imaging, 12 of the 16 CAD patients (75%) demonstrated evidence of myocardial ischemia. None of the 21 normal control subjects had evidence of mental stress induced myocardial ischemia. Mental stress also induced reproducible and significant hemodynamic changes in CAD patients. CONCLUSIONS: In patients with known CAD with typical angina or with evidence of reversible ischemia despite taking medications, mental stress was very effective in inducing myocardial ischemia, as detected by sestamibi SPECT imaging. Mental stress was also found to elicit significant hemodynamic responses. Furthermore, these findings demonstrated good reproducibility. PMID- 12569333 TI - Building a heart: implications for congenital heart disease. PMID- 12569334 TI - The genetics of congenital heart disease. PMID- 12569335 TI - Large animal models of congestive heart failure: a critical step in translating basic observations into clinical applications. AB - Congestive heart failure (CHF) is a clinical syndrome in which pathophysiologic underpinnings include left ventricular (LV) dysfunction, remodeling, and increased neurohormonal activation. Accordingly, large animal constructs must be developed that mimic this disease process in order to define new pharmacologic and surgical treatment strategies. Multiple large animal species have been used for these purposes. For instance, canine coronary artery microembolization has been used to generate ischemia-induced LV dilation and dysfunction. Sheep have been subjected to total acute coronary artery occlusion to evaluate ischemia induced mitral valve insufficiency. Rapid ventricular pacing has been used in both dogs and pigs to reproduce the characteristics of dilated cardiomyopathy. Each model is associated with advantages and disadvantages. Therefore findings derived from the study of large animal models of LV failure must be carefully evaluated. With proper interpretation, important insights into the pathogenesis of CHF may be realized. Furthermore, these models may be used in conjunction with imaging modalities such as magnetic resonance imaging, single photon emission computed tomography, and positron emission tomography to elucidate the identification of cellular and extracellular alterations associated with LV failure. Thus large animal models of CHF are critical components in the effort to translate basic observations into beneficial clinical applications. PMID- 12569336 TI - Risk stratification of the normal perfusion scan: does normal stress perfusion always mean very low risk? PMID- 12569337 TI - What lies beyond a chronically occluded coronary artery, and what should we do about it? PMID- 12569339 TI - Detection of pericardial effusion during Tc-99m sestamibi cardiac imaging. PMID- 12569340 TI - Comparing commercial attenuation correction methods for myocardial perfusion single photon emission computed tomography in phantoms. PMID- 12569338 TI - American Society of Nuclear Cardiology consensus statement: Task Force on Women and Coronary Artery Disease--the role of myocardial perfusion imaging in the clinical evaluation of coronary artery disease in women [correction]. PMID- 12569341 TI - Use of phantom studies to compare the performance of 8 different attenuation correction devices. PMID- 12569344 TI - Adjuvant systemic therapies for patients with breast cancer: endocrine responsiveness and effects of chemotherapy. PMID- 12569345 TI - Radiotherapy in early breast disease: current status and perspectives. PMID- 12569346 TI - Hereditary breast and ovarian cancer. PMID- 12569347 TI - News about the sentinel node in breast cancer. PMID- 12569348 TI - New possibilities of diagnostic imaging and intervention. PMID- 12569349 TI - How to treat good risk prostate cancer. PMID- 12569350 TI - Antiandrogen monotherapy for prostate cancer. PMID- 12569351 TI - Early or delayed hormonal therapy in N+ M+ disease. PMID- 12569352 TI - Adjuvant radiation therapy for rectal cancer: selecting the right cases. PMID- 12569353 TI - New potentials of radiotherapy in non-small cell lung cancer: stereotactic therapy and IMRT. PMID- 12569354 TI - Role of chemotherapy in the multimodality approach of non-small cell lung cancer. PMID- 12569355 TI - Modern management of symptoms and quality of life. PMID- 12569357 TI - An unusual H-Ras mutant isolated from a human multiple myeloma line leads to transformation and factor-independent cell growth. AB - Multiple myeloma (MM) is an incurable plasma cell malignancy. To investigate biochemical lesions associated with MM, we constructed an expression cDNA library from the OPM-2 human myeloma line. A highly transforming H-Ras mutant was identified by transfection analysis using NIH 3T3 cells. DNA sequencing demonstrated a single-point mutation at position 117 located in the guanine nucleotide-binding site resulting in a lysine-to-glutamic acid substitution. This mutant, H-Ras (K117E), was found to be constitutively activated in terms of GTP binding. We compared the biological effects of H-Ras (K117E) and H-Ras (G12V) in 32D murine hematopoietic progenitor cells. Whereas both Ras proteins are constitutively activated, 32D cells expressing H-Ras (G12V) are still dependent on IL-3 for survival and proliferation while cells carrying H-Ras (K117E) become IL-3 independent. Similar experiments conducted with the B9 line, an IL-6 dependent hybridoma, also demonstrated that B9/H-Ras (K117E) became IL-6 independent. Expression of H-Ras (K117E) in the human IL-6-dependent ANBL-6 myeloma line resulted in enhanced proliferation at suboptimal concentrations of IL-6. These observations suggest that H-Ras mutations at the binding site for the GTP nucleotide ring structure may also represent activating lesions and have additional biological effects when compared to previously described Ras mutants. PMID- 12569356 TI - hob1+, the fission yeast homolog of Bin1, is dispensable for endocytosis or actin organization, but required for the response to starvation or genotoxic stress. AB - BAR (Bin/Amphiphysin/Rvs) adapter proteins have been suggested to regulate endocytosis, actin organization, apoptosis, and transcription, but their precise roles are obscure. There are at least five mammalian genes that encode BAR adapter proteins, including the evolutionarily conserved and ubiquitously expressed Bin1/Amphiphysin-II and Bin3 genes. Bin1 holds special interest as certain splice isoforms localize to the nucleus, interact with the c-Abl and c Myc oncoproteins, and display tumor suppressor properties. To obtain functional insights, we embarked upon a genetic analysis of the two BAR adapter proteins expressed in the fission yeast Schizosaccharomyces pombe. In a previous work, a role in actin organization and cytokinesis was identified for the Bin3 homolog hob3+. In this study, a role in stress signaling was defined for the Bin1 homolog, hob1+. Notably, hob1+ was dispensable for endocytosis, actin organization, or osmotic sensitivity. Instead, mutation of hob1+ led to slight cell elongation and faulty cell cycle arrest upon nutrient starvation. These defects were complemented by Bin1, but not by Amphiphysin-I, arguing that these genes have distinct functions despite their structural similarity. hob1 delta mutant cells were also hypersensitive to genotoxic stress. This was not related to a faulty checkpoint response, but mutation in the checkpoint gene rad3(+) further exacerbated the sensitivity of hob1 delta mutant cells. Interestingly, mutation of the cell cycle regulator wee1+ partially relieved the sensitivity defect, suggesting that hob1+ may influence the efficiency of DNA repair or checkpoint release after DNA damage. Genetic and biochemical evidence indicated that hob3+ is epistatic to hob1+ in the response to genotoxic stress. Our findings indicate that the Bin1 homolog hob1+ participates in DNA damage signaling and they suggest a novel role for BAR adapter proteins in stress response processes. PMID- 12569358 TI - Effect of tyrosine kinase inhibitor STI571 on the kinase activity of wild-type and various mutated c-kit receptors found in mast cell neoplasms. AB - Systemic mastocytosis (SM) is a rare disease caused by an abnormal mast cell accumulation in various tissues. Two classes of constitutive activating c-kit mutations are found in SM. The most frequent class occurs in the catalytic pocket coding region with substitutions at codon 816 and the other in the intracellular juxtamembrane coding region. Therefore, kinase inhibitors that block mutated c kit activity might be used as therapeutic agents in SM. Here, we show that STI571 inhibits both wild-type and juxtamembrane mutant c-kit kinase activity, but has no effect on the activity of the D816 V mutant. Accordingly, STI571 selectively decreases the survival of normal mast cell and of mast cell lines either with juxtamembrane c-kit mutations, but not that of tumoral mast cell from patient with SM or of mast cell lines with the D816 V mutation. Therefore, STI571 is not a good candidate to treat SM and specific kinase inhibitors should be designed to inhibit constitutive activating mutations at codon 816. PMID- 12569359 TI - The twist gene is a common target of retroviral integration and transcriptional deregulation in experimental nephroblastoma. AB - The genes involved in the transformation of kidney blastema cells were searched for in avian nephroblastomas induced by the MAV2 retrovirus. The twist gene was identified as a common site of provirus integration in tumor cells. Twist was rearranged by the MAV2 provirus in three out of 76 independent nephroblastoma samples. The MAV2 integration sites were localized within 40 nucleotides of the twist 5'UTR region, right upstream from the ATG initiation codon. The integrated proviruses were deleted at their 5'ends. As a consequence, twist transcription became controlled by the retroviral 3'LTR promoter and was strongly upregulated, more than 200 times. In addition, 2-100 times elevated twist transcription was also detected in the majority of other nephroblastoma samples not containing MAV2 in the twist locus. We propose that chicken nephroblastoma originates from a single blastemic cell in which the MAV retrovirus, through its integration, has deregulated specific combinations of genes controlling proliferation and differentiation. The activation of the twist gene expression appears to contribute to tumorigenesis, as there is an in vivo positive selection of tumor cell clones containing the twist gene hyperactivated by MAV2 sequences inserted within the twist promoter. PMID- 12569360 TI - Expression of tetraspanins in human lung cancer cells: frequent downregulation of CD9 and its contribution to cell motility in small cell lung cancer. AB - Small cell lung cancer (SCLC) invades locally and metastasizes distantly extremely early when compared with nonsmall cell lung cancer (NSCLC). The underlying molecular mechanisms, however, have not been elucidated. Accumulating evidence suggests that downregulation of several members of tetraspanins is associated with progression of solid tumors, thus indicating poor prognosis. Here we screened 30 lung cancer cell lines for expression of tetraspanins, CD9, CD63, CD81, CD82, CD151, and NAG-2. Flow cytometry revealed that, among these proteins, CD9 is broadly expressed in NSCLC lines, but is absent or highly reduced in most SCLC lines (P<0.0001). Using the Boyden chamber and videomicroscopic cell motility assays, we showed that stable transfection of CD9 into an SCLC line, OS3 R5, reduced cell motility on fibronectin. Furthermore, by transient transfection of green fluorescent protein (GFP)-tagged CD9 into three other SCLC lines, we observed that SCLC cells expressing GFP-CD9 were uniformly less motile than untransfected cells. CD9 or GFP-CD9 was associated with beta1 integrins and distributed at the tumor cell periphery and cell-cell contacts, suggesting that CD9 modifies beta1 integrin function to reduce motility. These findings suggest that low expression of CD9 may contribute to the highly invasive and metastatic phenotype of SCLC. PMID- 12569361 TI - Bcl-x(L) and Akt cooperate to promote leukemogenesis in vivo. AB - To analyse individual factors that may contribute to leukemic transformation in vivo, we have developed a murine model of leukemogenesis based on the early hematopoietic precursor cell FL5.12. FL5.12 cells are interleukin-3 (IL-3) dependent for growth, proliferation, and survival. Relative resistance to cell death following IL-3 withdrawal can be conferred by either overexpression of the Bcl-x(L) apoptotic inhibitor, or constitutive activation of the serine/threonine kinase Akt. The ability of Bcl-x(L) or a constitutively active myristylated Akt to promote leukemic transformation of FL5.12 cells was compared in athymic nu(+)/nu(+) mice. Bcl-x(L) alone could not promote leukemic transformation, but mice injected with FL5.12 cells overexpressing Bcl-x(L) and a dominant-negative p53 construct developed leukocytosis and blastic infiltration of lymph nodes, spleen, and liver with features of a high-grade lymphoid malignancy. In contrast to the cells injected into these animals, cell lines derived from the mice were able to proliferate in the absence of IL-3, and were found to have constitutively activated Akt. This constitutive activation was associated with a variety of alterations of the signaling pathway regulating Akt activity, including alterations of PTEN mRNA and protein expression. In addition, some of these leukemic clones demonstrated concurrent constitutive upregulation of ERK activity. A constitutively active Akt construct introduced into FL5.12 cells promoted similar clonal expansion in vivo, with emergence of clonal IL-3 independent proliferation. Bcl-x(L) and Akt appeared to function cooperatively in this model, enhancing rapid clonal outgrowth in vivo relative to Akt alone. These results implicate activated Akt and growth-factor independence in leukemogenic transformation, and demonstrate the potential for in vivo analysis of genetic determinants of leukemogenesis. PMID- 12569362 TI - The MN1 oncoprotein synergizes with coactivators RAC3 and p300 in RAR-RXR mediated transcription. AB - The t(12;22) creates an MN1-TEL fusion gene leading to acute myeloid leukemia. The fusion partner TEL (ETV6) is a member of the ETS family of transcription factors. The nature of the other fusion partner, MN1, has not been investigated in detail until now. We recently described that MN1 activates the transcription activity of the moloney sarcoma virus long terminal repeat, indicating that this protein itself may act as a transcription factor. We show here that MN1 comprises multiple transcription activating domains. A search for a bound DNA sequence revealed that MN1 has affinity for retinoic acid responsive elements. A DR5 retinoic acid responsive element was observed in the LTR. The combination of MN1 and ligand-activated retinoic acid receptor leads to a synergistic induction of expression directed by the LTR. Cotransfection of MN1 with RAC3 or p300, known coactivators of retinoic acid receptors, leads to a further synergistic induction of transcription. In addition, the effect of MN1 can be inhibited by the wild type adenovirus ElA protein that inhibits p300 function, but not by an E1A mutant lacking the p300-binding site. GAL4-MN1-mediated transcription can be enhanced directly by RAC3 and p300. Taken together, our results indicate that MN1 is a transcription coactivator rather than a sequence-specific transcription factor, and that it may stimulate RAR/RXR-mediated transcription through interaction with p160 and p300. PMID- 12569364 TI - A human brain tumor-derived PDGFR-alpha deletion mutant is transforming. AB - Aberrant receptor tyrosine kinase signaling plays an important role in the molecular pathogenesis of brain tumors. We have been studying a previously identified human glioblastoma-derived PDGFR-alpha mutant that has an in-frame deletion in the extracellular domain, causing loss of exons 8 and 9 (PDGFR alpha(delta8,9)). In the primary tumor, this deletion mutant receptor was shown to be amplified and overexpressed. The purpose of this study was to determine the expression, activity, localization, and transformation properties of this deletion mutant. In the absence of serum, or PDGF-AA, PDGFR-alpha(delta8,9) was phosphorylated on tyrosine residues, indicating ligand-independent autoactivation. Localization by staining and cell surface biotinylation studies revealed expression of the deletion mutant predominantly in the cytoplasm, with very little present on the cell surface. To determine if PDGFR-alpha(delta8,9) was oncogenic, we transfected wild-type and mutant receptors into Rat1 cells and performed analyses of cell growth, in vitro transformation, and subcutaneous growth in the nude mouse. PDGFR-alpha(delta8,9)-expressing cells displayed enhanced cell growth and survival in low serum, and formed foci in monolayer cultures. PDGFR-alpha(delta8,9)-expressing Rat1 cells were also tumorigenic when injected subcutaneously into nude mice. Expression of PDGFR-alpha(delta8,9) was also associated with increased c-Jun phosphorylation in the absence of PDGF ligand, demonstrating also that the mutant receptor is associated with altered intracellular signaling. These data demonstrate that PDGFR-alpha(delta8,9) is transforming, and it is the first demonstration of a naturally occurring tumor derived mutant PDGFR-alpha with oncogenic properties. PMID- 12569363 TI - Selective PDZ protein-dependent stimulation of phosphatidylinositol 3-kinase by the adenovirus E4-ORF1 oncoprotein. AB - While PDZ domain-containing proteins represent cellular targets for several different viral oncoproteins, including human papillomavirus E6, human T-cell leukemia virus type 1 Tax, and human adenovirus E4-ORF1, the functional consequences for such interactions have not been elucidated. Here we report that, at the plasma membrane of cells, the adenovirus E4-ORF1 oncoprotein selectively and potently stimulates phosphatidylinositol 3-kinase (PI3K), triggering a downstream cascade of events that includes activation of both protein kinase B and p70S6-kinase. This activity of E4-ORF1 could be abrogated by overexpression of its PDZ-protein targets or by disruption of its PDZ domain-binding motif, which was shown to mediate complex formation between E4-ORF1 and PDZ proteins at the plasma membrane of cells. Furthermore, E4-ORF1 mutants unable to activate the PI3K pathway failed to transform cells in culture or to promote tumors in animals, and drugs that block either PI3K or p70S6-kinase inhibited E4-ORF1 induced transformation of cells. From these results, we propose that the transforming and tumorigenic potentials of the adenovirus E4-ORF1 oncoprotein depend on its capacity to activate PI3K through a novel PDZ protein-dependent mechanism of action. PMID- 12569365 TI - Overexpression of Cdc25B, an androgen receptor coactivator, in prostate cancer. AB - Cdc25B is a dual-specific phosphatase that mediates cell cycle progression by activating the cyclin-dependent kinases. It has been shown to possess oncogenic potential. To elucidate its potential contribution to human prostate cancer development, the expression profile of Cdc25B protein in human patients was analysed by immunohistocytochemistry. Cdc25B is frequently overexpressed in human prostate cancer tissues (29 of 30; 97%). In addition, the overexpression is more profound in the tumors of high combined Gleason scores and in late stages. Subsequently, we demonstrated that Cdc25B acts as a coactivator for AR in a hormone-dependent manner in the prostate cancer cell line, LNCaP. This coactivator function, surprisingly, is independent of its cell cycle functions. Cdc25B, on the other hand, directly interacts with AR as evidenced in GST-pull down and mammalian two-hybrid assays. In addition, it is also able to enhance AR mediated transcription in synergy with other coactivators, including CREB-binding protein (CBP) and p300/CBP associated factor. Therefore, upregulation of Cdc25B in human prostate cancer and its interplay with AR may contribute to prostate cancer development. PMID- 12569366 TI - The noncatalytic TrkCNC2 receptor is cleaved by metalloproteases upon neurotrophin-3 stimulation. AB - The trkC locus encodes catalytic and noncatalytic receptors, generated by alternative splicing. These primary high-affinity neurotrophin-3 (NT-3) receptors may act in concert to modulate responsiveness to NT-3. Signal modulation can also be achieved by receptors that are post-translationally processed. We report that the noncatalytic TrkC receptor, TrkCNC2, is cleaved at the membrane-proximal region of its extracellular domain. This generates a soluble ectodomain (gp90(TrkCNC2)) recovered in the cell culture medium and a membrane-bound fragment (p20(TrkCNC2)), which contains the transmembrane and intracellular regions including the juxtamembrane and the NC2-specific cytoplasmic domains. We also show that this processing, which does not occur in the TrkC catalytic counterpart, is upregulated by NT-3 and upon treatment with the tumor promoter 12 O-tetradecanoylphorbol-13-acetate. Moreover, cleavage inhibition after EDTA or 1.10 phenanthroline treatment suggests involvement of a metalloprotease(s). Finally, this post-translational processing was observed not only in TrkCNC2 overexpressing NIH3T3 cells but also in primary cultures of cortical neurons and brain extracts. This study shows that, in addition to alternative splicing, ectodomain shedding represents a novel means of regulating TrkC receptor signaling, and consequently NT-3 biological effects on target cells. PMID- 12569367 TI - Regulation of the ER81 transcription factor and its coactivators by mitogen- and stress-activated protein kinase 1 (MSK1). AB - The transcription factor ER81 has been shown to be involved in ontogenesis and breast tumor formation. ER81 is activated by many signals through phosphorylation directly mediated by mitogen-activated protein kinases (MAPKs), but also by an unknown protein kinase(s). Here, mitogen- and stress-activated protein kinase 1 (MSK1), which itself is directly activated by distinct classes of MAPKs, is identified to regulate ER81 function. MSK1 expression enhances ER81-dependent transcription upon stimulation of especially the p38-MAPK pathway. Two serine residues in ER81 are phosphorylated by MSK1, and mutating these serine residues to alanines dramatically diminishes the ability of MSK1 to stimulate ER81. However, mutation of the MSK1 phosphorylation sites in ER81 does not completely abrogate the ability of MSK1 to activate ER81 function, suggesting that MSK1 may also target cofactors of ER81. Consistently, MSK1 interacts with two homologous coactivators of ER81, CBP and p300, and stimulates the transactivation domains of CBP. Thus, MSK1 may regulate ER81-dependent transcription via direct phosphorylation of ER81 as well as via stimulation of CBP/p300, which might be important for ER81's normal function and during mammary tumor formation. PMID- 12569368 TI - Human HMGA2 promoter is coregulated by a polymorphic dinucleotide (TC)-repeat. AB - HMGA proteins are thought to be causally involved in the progression of different diseases, including benign and malignant tumors, obesity, arteriosclerosis, and restenosis. As HMGA proteins are architectural transcription factors, their binding to DNA leads to changes in DNA-conformation modulating the environment for the assembly and function of transcriptional complexes, thus influencing the expression of a huge variety of genes. Despite the emerging role of HMGA proteins for important diseases, only limited information is available about mechanisms regulating the expression of the HMGA2 gene. In this report, 2240 bp of the 5' flanking region of the HMGA2 gene were functionally analyzed by luciferase assay experiments. Besides the identification of novel positive and negative regulatory elements, it was shown that transcription is initiated from two independent promoter regions within cell lines HeLa, MCF7, and L14TSV40. Furthermore, a functional polymorphic dinucleotide repeat (TCTCT(TC)(n)) 500 bp upstream of the ATG translational start codon was found to regulate strongly the human HMGA2 promoter with an activation pattern that correlates to its TC-repeat length. PMID- 12569369 TI - Blockage of heregulin expression inhibits tumorigenicity and metastasis of breast cancer. AB - The growth factor heregulin (HRG), expressed in about 30% of breast cancer tumors, activates the erbB-2 receptor via induction of heterodimeric complexes of erbB-2 with erbB-3 or erbB-4. HRG induces tumorigenicity and metastasis of breast cancer cells. Our investigation into whether HRG is a factor likely to promote tumor formation independently of erbB-2 overexpression concludes that blockage of HRG expression suppresses the aggressive phenotype of MDA-MB-231 breast cancer cells by inhibiting cell proliferation, preventing anchorage-independent growth, and suppressing the invasive potential of the cells in vitro. More importantly, we observed a marked reduction in tumor formation, tumor size, and a lack of metastasis in vivo. These studies were achieved by blocking HRG expression in MDA MB-231 cells using an HRG antisense cDNA. In the search for the mechanism by which blockage of HRG reverts this aggressive phenotype, we discovered that the cells in which HRG is blocked exhibit a marked decrease in erbB activation and a significant reduction in MMP-9 activity, demonstrating a direct causal role in HRG induction of tumorigenicity. Our study is the first report and serves as a proof of the concept that HRG is a key promoter of breast cancer tumorigenicity and metastasis independently of erbB-2 overexpression and should be deemed a potential target in developing therapies for breast cancer. PMID- 12569370 TI - Cyclin E in breast tumors is cleaved into its low molecular weight forms by calpain. AB - Abundant levels of the hyperactive low molecular weight (LMW) forms of cyclin E contribute to deregulation of Cdk2 in breast tumors, but the mechanism through which they arise is not fully understood. Here, we explored the hypothesis that post-translational processing by a protease generates the LMW forms of cyclin E in breast tumors. In ZR75 tumor cell lysates, calcium-induced cyclin E truncation into peptides corresponding in size with LMW forms of cyclin E in tumor tissues. Calpeptin inhibited calcium-stimulated cyclin E truncation, indicating that cleavage resulted from activity of the calcium-dependent protease, calpain. Consistently, calcium+calpain caused truncation of cyclin E immunoprecipitated from tumor cells and tissues. Calcium also caused truncation of the calpain regulatory subunit in tumor cell lysates, indicating that elevated calpain activity accompanies cyclin E truncation. Increased levels of the calpain small subunit were also observed in breast tumors, and significant amounts of its proteolyzed forms indicated increased calpain activity. While elastase also caused cyclin E truncation, the cleavage pattern was distinct from that generated by calpain, suggesting discrete mechanisms in regulating the formation of LMW cyclin E in breast tumors. Treatment of ZR75 cultures with calcium+A23187 recapitulated the formation of the calcium/calpain-induced LMW forms of cyclin E. Altered calcium homeostasis and/or inability of the endogenous calpain inhibitor to control the activity of high levels of the calpain small subunit may contribute to increased calpain activity in breast tumors, causing abundant levels of LMW cyclin E. PMID- 12569371 TI - Tributyltin (TBT) induces ultra-rapid caspase activation independent of apoptosome formation in human platelets. AB - Activation of caspases has been demonstrated to be involved in thrombocytopenia and prolonged storage of platelet concentrates. Platelets represent enucleate cells that comprise all elements of the mitochondrial apoptosis pathway. However, no apoptotic stimuli capable of activating the endogenous caspase cascade have been identified so far. Using tributyltin (TBT) we could identify a compound that is capable of activating caspase-9 and -3 in platelets. Recent studies implicate that TBT induces apoptosis via the mitochondrial signaling pathway that is characterized by the formation of a high-molecular-weight complex (apoptosome) containing the adapter protein Apaf-1 and active caspase-9. Interestingly, addition of TBT induced the activation of caspase-9 in an ultra-rapid kinetic within the first 2 min. In addition, size exclusion chromatography revealed that TBT-mediated processing of caspase-9 occurs in the absence of the apoptosome. Thus, these data implicate that TBT induces the activation of caspase-9 by a mechanism not involving the formation of the apoptosome. PMID- 12569372 TI - ErbB-2 signaling is involved in regulating PSA secretion in androgen-independent human prostate cancer LNCaP C-81 cells. AB - The expression and secretion of prostate-specific antigen (PSA) are regulated by androgens in normal prostate secretory epithelial cells. In prostate cancer patients, the serum PSA level is usually elevated and cancer cells are initially responsive to androgens. However, those cancer cells become androgen-independent after androgen ablation therapy. In hormone-refractory cancer patients, even in an androgen-deprived environment, the circulation level of PSA rebounds and is constitutively elevated through a yet unknown mechanism. Tyrosine phosphorylation of ErbB-2 is involved in regulating the androgen-responsive phenotype of prostate cancer cells, and it is at least partly regulated by the cellular form of prostatic acid phosphatase (PAcP), a prostate-unique protein tyrosine phosphatase. We investigated the ErbB-2 signal pathway in androgen-independent PSA secretion. LNCaP C-81 cells, which are androgen-independent LNCaP cells lacking endogenous PAcP expression with a hypertyrosine phosphorylated ErbB-2, secreted a higher level of PSA in conditioned media than did androgen-sensitive LNCaP C-33 parental cells. A restored expression of cellular PAcP in C-81 cells was concurrent with a decrease in tyrophosphorylation of ErbB-2 and reduction of PSA secretion. Moreover, transient transfection of C-33 cells with the wild-type ErbB-2 or a constitutively active mutant of MEK1 cDNA resulted in an increased level of secreted PSA. The elevation of secreted PSA level by the forced expression of ErbB-2 was inhibited by an MEK inhibitor, PD98059. In C-81 cells, the expression of a dominant negative mutant of ErbB-2 reduced the secreted level of PSA. The inhibition of ErbB-2 or mitogen-activated protein (MAP) kinases by specific inhibitors AG879, AG825, or PD98059 led to a decrease in PSA secretion. Taken together, our data clearly indicate that the ErbB-2 signal pathway via MAP kinases (ERK1/2) is involved in regulating the secretion of PSA by androgen independent human prostate cancer LNCaP C-81 cells in an androgen-depleted environment. PMID- 12569373 TI - Randomised phase III trial of irinotecan combined with cisplatin for advanced non small-cell lung cancer. AB - To determine a standard combination chemotherapy for patients with advanced non small-cell lung cancer (NSCLC), we conducted a phase III trial of irinotecan (CPT 11) to test the hypotheses that CPT-11+cisplatin is superior to cisplatin+vindesine and that CPT-11 monotherapy is not inferior to cisplatin+vindesine. A total of 398 patients with previously untreated NSCLC were randomised to receive cisplatin+CPT-11 (CPT-P), cisplatin+vindesine (VDS-P) or CPT-11 alone (CPT). In the CPT-P arm, CPT-11 60 mg m(-2) was administered on days 1, 8 and 15, and cisplatin 80 mg m(-2) was administered on day 1. In the VDS-P arm, cisplatin 80 mg m(-2) was administered on day 1, and vindesine 3 mg m(-2) was administered on days 1, 8 and 15. In the CPT arm, CPT-11 100 mg m(-2) was administered on days 1, 8 and 15. The median survival time was 50.0 weeks for patients on CPT-P, 45.6 weeks for those on VDS-P and 46.0 weeks for those on CPT (P=0.115, CPT-P vs VDS-P; P=0.089, CPT vs VDS-P), and the hazard ratio was 0.85 (95% confidence interval (CI): 0.65-1.11) for CPT-P vs VDS-P and 0.83 (0.64-1.09) for CPT vs VDS-P. The response rate was 43.7% for patients on CPT-P, 31.7% for those on VDS-P and 20.5% for those on CPT. Major adverse reactions were grade 4 neutropenia observed in 37, 54 and 8% of the patients on CPT-P, VDS-P and CPT, respectively; and grades 3 and 4 diarrhoea observed in 12, 3 and 15% of the patients, respectively. CPT-P therapy produces comparable survival to VDS-P in patients with advanced NSCLC. CPT-11 monotherapy is not inferior to VDS-P in terms of survival. The CPT-11-containing regimen is one of the most efficacious and well tolerated in the treatment of advanced NSCLC. PMID- 12569375 TI - Metastatic renal carcinoma comprehensive prognostic system. AB - The purpose of the study was to identify a comprehensive prognostic system of pretreatment clinical parameters in 425 patients (pts) with metastatic renal-cell carcinoma treated with different subcutaneous (s.c.) recombinant cytokine-based home therapies in consecutive trials. Treatment consisted of (A) s.c. interferon alpha 2a (INF-alpha), s.c. interleukin-2 (IL-2) (n=102 pts), (B) s.c. IFN-alpha 2a, s.c. IL-2, and i.v. 5-fluorouracil (5-FU) (n=235 pts) or (C) s.c. IFN-alpha 2a, s.c. IL-2, and i.v. 5-FU combined with p.o. 13-cis-retinoic acid (13cRA) (n=88 pts). Kaplan-Meier survival analysis, log-rank statistics, and Cox regression analysis were employed to identify risk factors and to create a multiple risk factor model. The following pretreatment risk factors were identified by univariate analysis: (1) three and more metastatic sites, (2) presence of liver, lymph node or bone metastases, (3) neutrophil count > or = 6500 cells microl(-1), (4) serum lactate dehydrogenase level (LDH) > or = 220 U l(-1), and (5) serum C-reactive protein level (CRP) > or = 11 mg l(-1). Cox regression analysis with forward stepwise variable selection identified neutrophil count as the major prognostic factor (hazard ratio=1.9, P<0.001), while serum levels of LDH and CRP, time between diagnosis of tumour and onset of metastatic disease, number of metastatic sites, and bone metastases were significant but somewhat less important prognostic variables within the multiple risk factor model (hazard ratio < or = 1.5). Patients were assigned to one of the three risk groups according to cumulative risk defined as the sum of simplified risk s.c.ores for six pretreatment variables. Low-, intermediate-, and high-risk patients achieved a median overall survival of 32+ months (95% CI 24, 43; 5-year survival of 27%), 18+ months (95% CI 15, 20; 5-year survival of 11%), and 8+ months (95% CI 6, 10; 5-year survival of 5%), respectively. These prognostic categories are helpful both in individual patient care and in the assessment of patients entering prospective clinical trials. PMID- 12569374 TI - Gemcitabine and vinorelbine followed by docetaxel in patients with advanced non small-cell lung cancer: a multi-institutional phase II trial of nonplatinum sequential triplet combination chemotherapy (JMTO LC00-02). AB - To evaluate the efficacy and toxicity of the sequential nonplatinum combination chemotherapy consisting of gemcitabine (GEM) and vinorelbine (VNR) followed by docetaxel (DOC) in patients with advanced non-small-cell lung cancer (NSCLC), we conducted the multiinstitutional phase II study. A total of 44 chemotherapy-naive patients with advanced NSCLC were treated with GEM 1000 mg m(-2) and VNR 25 mg m( 2) intravenously on days 1 and 8 every 3 weeks for three cycles. DOC 60 mg m(-2) was then administrated intravenously at 3-week intervals for three cycles. Patients were evaluated for response and toxicity with each cycle of the treatment. The major objective response rate was 47.7% (95% confidence interval (CI), 33.8-62.1%). Median survival time (MST) was 15.7 months and 1-year survival rate was 59%. In the GEM/VNR cycle, grade 3/4 neutropenia occurred in 36.3%, grade 3/4 anaemia in two patients (4.5%) and grade 3 thrombocytopenia in one patient (2.3%). Grade 3 pneumonitis occurred in two patients (4.5%) in GEM/VNR cycles. In the DOC cycles, grade 3/4 neutropenia occurred in 39.4% but no patient experienced grade 3/4 anaemia or thrombocytopenia. Of the 44 eligible patients, 33 patients completed three cycles of GEM/VNR and 22 patients completed six cycles of planned chemotherapy (three cycles of GEM/VNR followed by three cycles of DOC). The sequential triplet nonplatinum chemotherapy consisted of GEM/VNR followed by DOC, and was very active and well tolerated. This study forms the basis for an ongoing phase III trial that compares this nonplatinum triplet and standard platinum doublet combination (carboplatin/paclitaxel). PMID- 12569376 TI - Rapid access multidisciplinary lymph node diagnostic clinic: analysis of 550 patients. AB - Lymphadenopathy is common, affecting patients of all ages. The current referral pattern for investigating patients with lymphadenopathy varies widely with no universally practised pathway. Our institution set up a lymph node diagnostic clinic (LNDC) accepting direct referrals from primary care physicians. Details of clinical presentation and investigations were recorded prospectively. Between December 1996 and July 2001, 550 patients were referred (M: 203; F:347). The median age was 40 years (range 14-90). The median time between initial referral and the first clinic visit was 6 days. Of 95 patients diagnosed to have malignant diseases, the median time from the first clinic visit to reaching malignant diagnosis was 15 days. Multivariate logistic regression analysis identified five significant predictors for malignant nodes: male gender (risk ratio (RR)=2.72; 95% confidence interval (CI): 1.63-4.56), increasing age (RR=1.05; 95% CI: 1.04 1.07), white ethnicity (RR=3.01; 95% CI: 1.19-7.6) and sites of lymph nodes: supraclavicular region (RR=3.72; 95% CI: 1.52-9.12) and > or = 2 regions of lymph nodes (RR=6.41; 95% CI: 2.82-14.58). Ultrasound and fine-needle aspiration cytology of palpable lymph nodes were performed in 154 and 289 patients, respectively. An accuracy of 97 and 84% was found, respectively. In conclusion, a multidisciplinary lymph node diagnostic clinic enables a rapid, concerted approach to a common medical problem and patients with malignant diseases were diagnosed in a timely fashion. PMID- 12569377 TI - Breast cancer incidence after the start of mammography screening in Denmark. AB - Mammography screening may lead to overdiagnosis of asymptomatic breast cancers, that would otherwise not have given rise to clinical symptoms. This aspect was studied in three regional screening programmes in Denmark, which started in Copenhagen municipality, Fyn county, and Frederiksberg municipality in 1991, 1993, and 1994, respectively. In these regions, we compared time trends in incidence of invasive breast cancer with the rest of Denmark. Since the number of clinical mammograms was relatively low, it was reasonable to assume that the breast cancer incidence outside the three screening regions represented the incidence of a population with low-intensity opportunistic screening. In Copenhagen and Fyn, a prevalence peak in incidence was seen during the first invitation round. During the subsequent invitation rounds, the incidence dropped to a level in line with the incidence expected without screening. The pattern was different in the small municipality of Frederiksberg, where the sensitivity was low during the first invitation round. Inclusion of screen-detected ductal carcinoma in situ cases did not change these results. The experiences from Copenhagen and Fyn show that organised mammography screening can operate without overdiagnosis of breast cancer. PMID- 12569378 TI - The precancer risk of betel quid chewing, tobacco use and alcohol consumption in oral leukoplakia and oral submucous fibrosis in southern Taiwan. AB - In areas where the practise of betel quid chewing is widespread and the chewers also often smoke and drink alcohol, the relation between oral precancerous lesion and condition to the three habits is probably complex. To explore such association and their attributable effect on oral leukoplakia (OL) and oral submucous fibrosis (OSF), a gender-age-matched case-control study was conducted at Kaohsiung, southern Taiwan. This study included 219 patients with newly diagnosed and histologically confirmed OL or OSF, and 876 randomly selected community controls. All information was collected by a structured questionnaire through in-person interviews. A preponderance of younger patients had OSF, while a predominance of older patients had OL. Betel quid chewing was strongly associated with both these oral diseases, the attributable fraction of OL being 73.2% and of OSF 85.4%. While the heterogeneity in risk for areca nut chewing across the two diseases was not apparent, betel quid chewing patients with OSF experienced a higher risk at each exposure level of chewing duration, quantity and cumulative measure than those who had OL. Alcohol intake did not appear to be a risk factor. However, cigarette smoking had a significant contribution to the risk of OL, and modified the effect of chewing based on an additive interaction model. For the two oral premalignant diseases combined, 86.5% was attributable to chewing and smoking. Our results suggested that, although betel quid chewing was a major cause for both OL and OSF, its effect might be difference between the two diseases. Cigarette smoking has a modifying effect in the development of oral leukoplakia. PMID- 12569379 TI - Parental smoking and childhood cancer: results from the United Kingdom Childhood Cancer Study. AB - There are strong a priori reasons for considering parental smoking behaviour as a risk factor for childhood cancer but case - control studies have found relative risks of mostly only just above one. To investigate this further, self-reported smoking habits in parents of 3838 children with cancer and 7629 control children included in the United Kingdom Childhood Cancer Study (UKCCS) were analysed. Separate analyses were performed for four major groups (leukaemia, lymphoma, central nervous system tumours and other solid tumours) and more detailed diagnostic subgroups by logistic regression. In the four major groups, after adjustment for parental age and deprivation there were nonsignificant trends of increasing risk with number of cigarettes smoked for paternal preconception smoking and nonsignificant trends of decreasing risk for maternal preconception smoking (all P-values for trend >0.05). Among the diagnostic subgroups, a statistically significant increased risk of developing hepatoblastoma was found in children whose mothers smoked preconceptionally (OR=2.68, P=0.02) and strongest (relative to neither parent smoking) for both parents smoking (OR=4.74, P=0.003). This could be a chance result arising from multiple subgroup analysis. Statistically significant negative trends were found for maternal smoking during pregnancy for all diagnoses together (P<0.001) and for most individual groups, but there was evidence of under-reporting of smoking by case mothers. In conclusion, the UKCCS does not provide significant evidence that parental smoking is a risk factor for any of the major groups of childhood cancers. PMID- 12569380 TI - Sex ratio among offspring of childhood cancer survivors treated with radiotherapy. AB - It has been postulated that paternal gonadal exposure would increase the sex ratio by inducing X-chromosomal dominant lethals but that maternal gonadal exposure would decrease the sex ratio by inducing recessive sex-linked lethals. We therefore evaluated the sex ratio (male-to-female ratio) of children born to survivors of childhood cancers in Denmark. Children with cancer were identified from the Danish Cancer Registry from 1943 to 1996 and their offspring from the Central Population Registry. Radiation treatments were determined from records within the Cancer Registry and gonadal radiation exposures were estimated based on the cancer being treated and the likely proximity of the radiation fields to the gonads. Overall, 1100 survivors of childhood cancer became the parents of 2130 children. The sex ratio for male (0.99) and female (1.00) cancer survivors was similar and did not differ significantly from the Danish population (1.06). Radiotherapy did not influence the sex ratio of the children of either male or female survivors, and there was no evidence for dose-related changes over categories of estimated dose to parental gonads. We saw no consistent association between the sex ratio and the interval between cancer diagnosis of the parent and birth of the child. This nationwide study provides no support for the hypothesis that radiation exposure to the gonads results in an inherited genetic effect that would be manifested by a change in the sex ratio of children born after exposure. It may be, however, that sex ratio alterations are not a good or even a valid indicator of possible genetic effects in humans. PMID- 12569381 TI - Cell cycle checkpoint status in human malignant mesothelioma cell lines: response to gamma radiation. AB - Knowledge of the function of the cell cycle checkpoints in tumour cells may be important to develop treatment strategies for human cancers. The protein p53 is an important factor that regulates cell cycle progression and apoptosis in response to drugs. In human malignant mesothelioma, p53 is generally not mutated, but may be inactivated by SV40 early region T antigen (SV40 Tag). However, the function of p53 has not been investigated in mesothelioma cells. Here, we investigated the function of the cell cycle checkpoints in six human mesothelioma cell lines (HMCLs) by studying the cell distribution in the different phases of the cell cycle by flow cytometry, and expression of cell cycle proteins, p53, p21(WAF1/CIP1) and p27(KIP1). In addition, we studied p53 gene mutations and expression of SV40 Tag. After exposure to gamma-radiation, HMCLs were arrested either in one or both phases of the cell cycle, demonstrating a heterogeneity in cell cycle control. G1 arrest was p21(WAF1/CIP1)- and p53-dependent. Lack of arrest in G1 was not related to p53 mutation or binding to SV40 Tag, except in one HMCL presenting a missense mutation at codon 248. These results may help us to understand mesothelioma and develop new treatments. PMID- 12569382 TI - ErbB2 and bone sialoprotein as markers for metastatic osteosarcoma cells. AB - Osteosarcoma is the most common malignant bone neoplasia occurring in young patients in the first two decades of life, and represents 20% of all primitive malignant bone tumours. At present, treatment of metastatic osteosarcoma is unsatisfactory. High-dose chemotherapy followed by CD34+ leukapheresis rescue may improve these poor results. Neoplastic cells contaminating the apheresis may, however, contribute to relapse. To identify markers suitable for detecting osteosarcoma cells in aphereses we analysed the expression of bone-specific genes (Bone Sialoprotein (BSP) and Osteocalcin) and oncogenes (Met and ErbB2) in 22 patients with metastatic osteosarcoma and six healthy stem cell donors. The expression of these genes in aphereses of patients affected by metastatic osteosarcoma was assessed by RT-PCR and Southern blot analysis. Met and Osteocalcin proved to be not useful markers since they are positive in aphereses of both patients with metastatic osteosarcoma and healthy stem cell donors. On the contrary, BSP was expressed at significant levels in 85% of patients. Moreover, 18% of patients showed a strong and significantly positive (seven to 16 times higher than healthy stem cell donors) ErbB2 expression. In all positive cases, neoplastic tissue also expressed ErbB2. Our data show that ErbB2 can be a useful marker for tumour contamination in aphereses of patients affected by ErbB2 expressing osteosarcomas and that analysis of Bone Sialoprotein expression can be an alternative useful marker. PMID- 12569383 TI - Prognostic value of Goseki histological classification in adenocarcinoma of the cardia. AB - Various histologic classification systems have been proposed as prognostic factors for gastric cancer. We assessed the prognostic value of Goseki classification as well as the TNM staging system, histological tumour grading, Lauren, WHO, Goseki and Siewert classifications in 100 patients with cardia carcinoma undergoing curative surgery. Two patients were lost at follow-up. The median time of follow-up in the remaining patients was 32.9 months after surgery (range: 0.1-142.1 months). No differences in survival rates were observed according to tumour grading, Lauren or WHO histologic or Siewert topographical classification. No differences were found according to Goseki classes, when considering either the mucin content of the carcinoma (types I and III vs II and IV) or the differentiation grade (types I and II vs III and IV). Multivariate analysis showed that the only lymph node positivity was a significant predictor of survival: 7.2% of patients with, but 41.5% of those without nodal involvement were alive after five years (P=0.0001). In conclusion, we found no prognostic role for Goseki or the traditional histological indexes, while the TNM staging system and particularly lymph node positivity were the main predictors of survival in patients with cardia adenocarcinoma. PMID- 12569384 TI - Breast cancer response to neoadjuvant chemotherapy: predictive markers and relation with outcome. AB - The aim of this study was to provide a better insight into breast cancer response to chemotherapy. Chemotherapy improves outcome in breast cancer patients. The effect of cytotoxic treatment cannot be predicted for individual patients. Therefore, the identification of tumour characteristics associated with tumour response and outcome is of great clinical interest. We studied 97 patients, who received anthracycline-based neoadjuvant chemotherapy. Tumour samples were taken prior to and after chemotherapy. We quantified the response to chemotherapy clinically and pathologically and determined histological and molecular tumour characteristics. We assessed changes in the expression of Bcl-2, ER, P53 HER2 and Ki-67. Association with response and outcome was tested for all parameters. The experimental results showed 15 clinical (17%) and three (3%) pathological complete remissions. There were 18 (20%) clinical vs 29 (33%) pathological nonresponders. The expression of most markers was similar before and after chemotherapy. Only Ki-67 was significantly decreased after chemotherapy. Factors correlated with response were: large tumour size, ER negativity, high Ki-67 count and positive P53 status. Tumour response and marker expression did not predict disease-free or overall survival. In conclusion, clinical and pathological response assessments are poorly associated. Proliferation decreases significantly after chemotherapy. ER negativity and a high proliferation index are associated with better response. HER2 status does not predict response, and outcome is not related to tumour response. PMID- 12569385 TI - CpG island methylation is a common finding in colorectal cancer cell lines. AB - Tumour cell lines are commonly used in colorectal cancer (CRC) research, including studies designed to assess methylation defects. Although many of the known genetic aberrations in CRC cell lines have been comprehensively described, no studies have been performed on their methylation status. In this study, 30 commonly used CRC cell lines as well as seven primary tumours from individuals with hereditary nonpolyposis colorectal cancer (HNPCC) were assessed for methylation at six CpG islands known to be hypermethylated in colorectal cancer: hMLH1, p16, methylated in tumour (MINT-)-1, -2, -12 and -31. The cell lines were also assessed for microsatellite instability (MSI), ploidy status, hMLH1 expression, and mutations in APC and Ki-ras. Methylation was frequently observed at all examined loci in most cell lines, and no differences were observed between germline-derived and sporadic cell lines. Methylation was found at MINT 1 in 63%, MINT 2 in 57%, MINT 12 in 71%, MINT 31 in 53%, p16 in 71%, and hMLH1 in 30% of cell lines. Overall only one cell line, SW1417, did not show methylation at any locus. Methylation was found with equal frequency in MSI and chromosomally unstable lines. MSI was over-represented in the cell lines relative to sporadic CRC, being detected in 47% of cell lines. The rate of codon 13 Ki-ras mutations was also over three times that expected from in vivo studies. We conclude that CpG island hypermethylation, whether acquired in vivo or in culture, is a ubiquitous phenomenon in CRC cell lines. We suggest that CRC cell lines may be only representative of a small subset of real tumours, and this should be taken into account in the use of CRC cell lines for epigenetic studies. PMID- 12569386 TI - Infrequent mutation of the tumour-suppressor gene Smad4 in early-stage colorectal cancer. AB - Smad4 is a candidate tumour-suppressor gene identified recently on chromosome 18q21.1. Both alleles are inactivated in nearly one-half of pancreatic carcinomas, but its role in the tumorigenesis of other tumours is still unknown. The aim of this study was to investigate the potential involvement of the Smad4 locus in early-stage colorectal cancers (stages I-III) in tumour samples from a randomised multicentre trial. Of a large collection of DNA samples, 73 with a loss of one allele of the Smad4 gene were analysed for the presence of point mutations in the remaining gene. Patients, from whom biopsies were isolated, were part of a previous randomised multicentre study of the Swiss Group for Clinical Cancer Research on the benefit of adjuvant chemotherapy (SAKK study 40/81). Mutation analysis was restricted to the highly conserved C-terminal domain (exons 8, 9, 10 and 11) of Smad4, using PCR and single-strand conformational variant analysis. Two of the 73 patients (3%) with loss of one allele of Smad4 had a point mutation in the remaining allele. These results indicate that whereas Smad4 point mutations are prevalent in pancreatic carcinoma, they are infrequent in early stages (I-III) of colorectal cancer. PMID- 12569387 TI - On the biological relevance of MHC class II and B7 expression by tumour cells in melanoma metastases. AB - A large number of studies have indicated that specific immune reactivity plays a crucial role in the control of malignant melanoma. In this context, expression of MHC I, MHC II and B7 molecules by melanoma cells is seen as relevant for the immune response against the tumour. For a better understanding of the biological relevance of MHC II and B7 expression by tumour cells in metastatic melanoma, we studied the expression of these molecules in melanoma metastases in relation to the inflammatory response, regression of the tumour and survival from 27 patients treated with biochemotherapy (30 mg m(-2) Cisplatin and 250 mg m(-2) decarbazine (dimethyl-triazene-imidazole-carboxamide, DTIC) on days 1-3 i.v., and 10(7) IU IFN-alpha 2b 3 days a week s.c., q. 28d). In 19 out of 27 lesions studied, we found expression of MHC II by the tumour cells, while only in one out of 11 tumour biopsies obtained from untreated metastatic melanoma patients, MHC II expression was detected. Expression of B7.1 and B7.2 by tumour cells was found in nine out of 24 and 19 out of 24 lesions, respectively. In all cases where B7.1 expression was found, expression of B7.2 by the tumour cells was also seen. In general, no or only few inflammatory cells positive for B7 were found. Expression of MHC II by tumour cells was positively correlated with the presence of tumour infiltrating lymphocytes, regression of the lesion, and with time to progression (TTP) and overall survival (OS) of the patient. However, no significant correlation between B7.1 or B7.2 expression and regression of the tumour, TTP or OS was found. In light of other recent findings, these data altogether do support a role as biomarker for MHC II expression by tumour cells; however, its exact immunological pathomechanism(s) remain to be established. PMID- 12569388 TI - Topical application of ALA and ALA hexyl ester on a subcutaneous murine mammary adenocarcinoma: tissue distribution. AB - Although 5-aminolevulinic acid (ALA)-based photodynamic therapy (PDT) has proven to be clinically beneficial for the treatment of certain cancers, including a variety of skin cancers, optimal tissue localisation still remains a problem. An approach to improve the bioavailability of protoporphyrin IX (PpIX) is the use of ALA derivatives instead of ALA. In this work, we employed a subcutaneous murine mammary adenocarcinoma to study the tissue distribution pattern of the ALA hexyl ester (He-ALA) in comparison with ALA after their topical application in different vehicles. He-ALA induced porphyrin synthesis in the skin overlying the tumour (SOT), but it did not reach the tumour tissue as efficiently. Only 5 h after He-ALA lotion application, tumour porphyrin levels surpassed control values. He-ALA delivered in cream induced a substantially lower porphyrin synthesis in SOT, reinforcing the importance of the vehicle in the use of topical PDT. Porphyrin levels in internal organs remained almost within control values when He-ALA was employed. The addition of DMSO to ALA formulation slightly increased tumour and SOT porphyrin biosynthesis, but it did not when added to He ALA lotion. PMID- 12569389 TI - SR31747A is a sigma receptor ligand exhibiting antitumoural activity both in vitro and in vivo. AB - SR31747A is a recently described sigma receptor ligand that binds SR31747A binding protein 1 (SR-BP) and emopamil-binding protein (EBP) (also called the sigma 1 receptor and the human sterol isomerase (HSI), respectively), and has immunoregulatory and antiproliferative activities. To further investigate its antitumour activity and focusing on cancers, which are sensitive to the molecule, we measured the proliferation of different human epithelial breast or prostate cancer cell lines following in vitro and in vivo SR31747A treatment. Firstly, in vitro, we found that nanomolar concentrations of SR31747A dramatically inhibited cell proliferation in both hormono-responsive and -unresponsive cancer cell lines. Secondly, tumour development was significantly decreased in mice treated with SR31747A. In an attempt to decipher the SR31747A mode of action, we found that the two binding sites may not fully account for this activity. Indeed, while competitive experiments indicated that EBP prevails in mediating SR31747A antiproliferative activity, an analysis of the expression of both receptors indicated that the cellular sensitivity to SR31747A is not correlated with either EBP or SR-BP expression. These data suggest that additional binding sites may exist. Preliminary binding studies demonstrated that SR31747A also binds to sigma 2, a protein that has not yet been cloned, but which is considered as a potential marker of the proliferative status of tumour cells. Altogether, our data demonstrate the antitumoural activity of SR31747A both in vitro and in vivo in two different cancer models, broaden the spectrum of its binding proteins and enhance the potential for further therapeutic development of the molecule. PMID- 12569390 TI - Changes in the ornithine cycle following ionising radiation cause a cytotoxic conditioning of the culture medium of H35 hepatoma cells. AB - Cultured H35 hepatoma cells release a cytotoxic factor in response to irradiation with X-rays. When the conditioned medium from irradiated cells is given to nonirradiated cells, growth is inhibited and followed by cell death, possibly apoptosis, Analysis of the conditioned medium reveals a dramatic change in the ornithine (urea) cycle components after the irradiation. A strong decrease in medium arginine is accompanied with parallel increases in ornithine, citrulline and ammonia. The high level of ammonia appears to be largely responsible for the observed cytotoxicity. The development of hyperammonia by irradiated cells and the related toxicity depend on the radiation dose and the number of cells seeded thereafter for the medium conditioning. Development of cytotoxicity by irradiated cells is completely prevented with the arginase inhibitor L-norvaline, in arginine-deficient medium or when citrulline replaces arginine. These preventive measures result in subtoxic ammonia levels. PMID- 12569391 TI - Effect of CD26/dipeptidyl peptidase IV on Jurkat sensitivity to G2/M arrest induced by topoisomerase II inhibitors. AB - CD26/dipeptidyl peptidase IV (DPPIV) is a surface antigen with multiple functions, including a role in T-cell activation and the development of certain human cancers. We previously demonstrated that CD26/DPPIV enhanced sensitivity of Jurkat cells to doxorubicin. We now show that expression of CD26/DPPIV enhanced sensitivity of CD26 Jurkat transfectants to G(2)-M arrest mediated by the antineoplastic agent etoposide. The increased sensitivity to etoposide-induced G(2)-M arrest was associated with disruption of cell cycle-related events, including hyperphosphorylation of p34(cdc2) kinase, change in cdc25C expression and phosphorylation, and alteration in cyclin B1 expression. CD26/DPPIV associated enhancement of doxorubicin and etoposide-induced G(2)-M arrest was also observed in serum-free media, suggesting an effect of CD26 on cell-derived processes rather than serum-derived factors. Importantly, our work elucidated a potential mechanism for the enhanced susceptibility of CD26-expressing Jurkat cells to the topoisomerase II inhibitors by demonstrating that CD26/DPPIV surface expression was associated with increased topoisomerase II alpha levels and enhanced enzyme activity. Besides being the first to show a functional association between the multifaceted molecule CD26 and the key cellular protein topoisomerase II alpha, our studies provide additional evidence of a potential role for CD26 in the treatment of selected malignancies. PMID- 12569392 TI - Temozolomide induces apoptosis and senescence in glioma cells cultured as multicellular spheroids. AB - Temozolomide is an alkylating cytostatic drug that finds increasing application in the treatment of melanoma, anaplastic astrocytoma and glioblastoma multiforme. The compound is a prodrug that decomposes spontaneously, independent of an enzymatic activation step. DNA methylation induces futile mismatch repair cycles and depletion of the DNA repair enzyme O(6)-methylguanine-DNA methyltransferase should then initiate programmed cell death. We show drug-dependent inhibition of tumour growth in a three-dimensional cell culture model of the glioma cell lines U87MG and GaMG. Migrational behaviour of the glioblastoma cells remained unaltered. However, coincubation of tumour spheroids with primary brain aggregates showed reduced tumour cell invasion into brain tissue in the presence of temozolomide. This was not achieved by slowing cellular migration, as temozolomide-treated cells displayed no reduced motility. By transferase-mediated dUTP nick-end labelling (TUNEL) of apoptotic nuclei, we found that the drug was able to induce apoptosis throughout the tumour cell spheroids. Apoptosis was highest in the core region of the spheroids. Repetitive application of sublethal doses of temozolomide to multicellular spheroids resulted in the development of drug resistance in GaMG cells. We suggest that temozolomide is a strong initiator of apoptosis in glioblastoma tumour cells in a spheroid cell culture system, when cells are already in a stressful environment. PMID- 12569393 TI - Antitumour 2-(4-aminophenyl)benzothiazoles generate DNA adducts in sensitive tumour cells in vitro and in vivo. AB - 2-(4-Aminophenyl)benzothiazoles represent a potent and highly selective class of antitumour agent. In vitro, sensitive carcinoma cells deplete 2-(4 aminophenyl)benzothiazoles from nutrient media; cytochrome P450 1A1 activity, critical for execution of antitumour activity, and protein expression are powerfully induced. 2-(4-Amino-3-methylphenyl)benzothiazole-derived covalent binding to cytochrome P450 1A1 is reduced by glutathione, suggesting 1A1 dependent production of a reactive electrophilic species. In vitro, 2-(4 aminophenyl)benzothiazole-generated DNA adducts form in sensitive tumour cells only. At concentrations >100 nM, adducts were detected in DNA of MCF-7 cells treated with 2-(4-amino-3-methylphenyl)-5-fluorobenzothiazole (5F 203). 5F 203 (1 microM) led to the formation of one major and a number of minor adducts. However, treatment of cells with 10 microM 5F 203 resulted in the emergence of a new dominant adduct. Adducts accumulated steadily within DNA of MCF-7 cells exposed to 1 microM 5F 203 between 2 and 24 h. Concentrations of the lysylamide prodrug of 5F 203 (Phortress) > or = 100 nM generated adducts in the DNA of sensitive MCF 7 and IGROV-1 ovarian cells. At 1 microM, one major Phortress-derived DNA adduct was detected in these two sensitive phenotypes; 10 microM Phortress led to the emergence of an additional major adduct detected in the DNA of MCF-7 cells. Inherently resistant MDA-MB-435 breast carcinoma cells incurred no DNA damage upon exposure to Phortress (< or = 10 microM, 24 h). In vivo, DNA adducts accumulated within sensitive ovarian IGROV-1 and breast MCF-7 xenografts 24 h after treatment of mice with Phortress (20 mg kg(-1)). Moreover, Phortress derived DNA adduct generation distinguished sensitive MCF-7 tumours from inherently resistant MDA-MB-435 xenografts implanted in opposite flanks of the same mouse. PMID- 12569394 TI - Noninvasive estimation of tumour viability in a xenograft model of human neuroblastoma with proton magnetic resonance spectroscopy (1H MRS). AB - The aim of the study was to evaluate proton magnetic resonance spectroscopy ((1)H MRS) for noninvasive biological characterisation of neuroblastoma xenografts in vivo. For designing the experiments, human neuroblastoma xenografts growing subcutaneously in nude rats were analysed in vivo with (1)H MRS and magnetic resonance imaging at 4.7 T. The effects of spontaneous tumour growth and antiangiogenesis treatment, respectively, on spectral characteristics were evaluated. The spectroscopic findings were compared to tumour morphology, proliferation and viable tumour tissue fraction. The results showed that signals from choline (Cho)-containing compounds and mobile lipids (MLs) dominated the spectra. The individual ML/Cho ratios for both treated and untreated tumours were positively correlated with tumour volume (P<0.05). There was an inverse correlation between the ML/Cho ratio and the viable tumour fraction (r=-0.86, P<0.001). Higher ML/Cho ratios concomitant with pronounced histological changes were seen in spectra from tumours treated with the antiangiogenic drug TNP-470, compared to untreated control tumours (P<0.05). In conclusion, the ML/Cho ratio obtained in vivo by (1)H MRS enabled accurate assessment of the viable tumour fraction in a human neuroblastoma xenograft model. (1)H MRS also revealed early metabolic effects of antiangiogenesis treatment. (1)H MRS could prove useful as a tool to monitor experimental therapy in preclinical models of neuroblastoma, and possibly also in children. PMID- 12569395 TI - Iron uptake, trafficking and homeostasis in plants. AB - Iron is an essential micronutrient with numerous cellular functions, and its deficiency represents one of the most serious problems in human nutrition worldwide. Plants have two major problems with iron as a free ion: its insolubility and its toxicity. To ensure iron acquisition from soil and to avoid iron excess in the cells, uptake and homeostasis are tightly controlled. Plants meet the extreme insolubility of oxidized iron at neutral pH values by deficiency inducible chelation and reduction systems at the root surface that facilitate uptake. Inside the cells the generation of highly toxic hydroxyl radicals by iron redox changes is avoided by intricate chelation mechanisms. Organic acids, most notably nicotianamine, and specialized proteins bind iron before it can be inserted into target molecules for biological function. Uptake and trafficking of iron throughout the plant is therefore a highly integrated process of membrane transport and reduction, trafficking between chelator species, whole-plant allocation and genetic regulation. The improvement of crop plants with respect to iron efficiency on iron-limiting soils and to iron fortification for human nutrition has been initiated by breeding and biotechnology. These efforts have to consider molecular and physiological evidence to overcome the inherent barriers and problems of iron metabolism. PMID- 12569396 TI - Differential ozone sensitivity among Arabidopsis accessions and its relevance to ethylene synthesis. AB - We compared the physiological and molecular responses of two Arabidopsis accessions, Col-0 and Ws-2, to ozone (O(3)) exposure. Observation of visible injury as well as ion-leakage analysis demonstrated clear differences between the O(3)-tolerant accession Col and the O(3)-sensitive accession Ws. RNA-blot analysis showed that O(3)-induced increases in mRNA levels of several ethylene inducible genes and a salicylic acid-inducible gene were substantially higher in Ws than in Col. The time-course of induction of various mRNA levels shows that the expression of ethylene-inducible genes was rapidly, and more strongly, induced by O(3) in Ws than in Col, suggesting that Ws exhibits higher ethylene signaling. Both the level of mRNA for an O(3)-inducible 1-aminocyclopropane-1 carboxylate synthase and the level of ethylene generation after 3 h of O(3) exposure were higher in Ws than in Col. O(3)-induced leaf damage was attenuated by pretreatment with ethylene biosynthesis- and signaling-inhibitors, indicating that ethylene signaling is required for O(3)-induced leaf injury in Ws. On the other hand, an ethylene-overproducing mutant of Col, eto1-1, displayed significantly increased O(3)-induced leaf injury compared to wild type plants. These results indicate that the difference in O(3) sensitivity is dependent on the difference in ethylene production rate between these two accessions. Finally, we investigated the relationship between the degree of leaf damage and the level of ethylene evolution in 20 different Arabidopsis accessions. Based on the result, the accessions were classified into four types. However, most of them showed significant correlation between the ethylene production level and the degree of leaf injury, suggesting that ethylene signaling is an important factor in the natural variety of O(3) sensitivity among Arabidopsis accessions. PMID- 12569397 TI - Regulation of biosynthesis and intracellular localization of rice and tobacco homologues of nucleosome assembly protein 1. AB - The nucleosome assembly protein 1 (NAP1) is considered to be a conserved histone chaperone, facilitating the assembly of nucleosomes in all eukaryotes. However, studies in yeast and animal cells also indicated that NAP1 proteins have diverse functions likely independent of nucleosome-assembly activity. Here, we describe the isolation and characterization of cDNAs encoding NAP1-like proteins from the monocotyledon rice ( Oryza sativa L.) and the dicotyledon tobacco ( Nicotiana tabacum L.). Northern-blot analysis demonstrated that the two rice NAP1-like genes are predominantly expressed in stem tissues such as root and shoot apical meristems as well as in young flowers. During the cell cycle, all four tobacco NAP1-like genes are highly expressed, with one of them showing a slightly increased expression at the G1/S transition. These results are consistent with a role for plant NAP1-like proteins in cell division. In vitro binding assays revealed that different NAP1-like proteins bind, with distinct relative binding strengths, to different classes of histone. Intracellular localization analyses showed that some NAP1-like proteins could be targeted into the nucleus whereas others are exclusively cytoplasm-localized. It is thus likely that different plant NAP1-like proteins have distinct functions in vivo. Plant NAP1-like proteins were observed to concentrate around the metaphase plate and in the phragmoplast, suggesting a role in mitotic events and cytokinesis. PMID- 12569398 TI - Stomatal neighbor cell polarity and division in Arabidopsis. AB - Asymmetric divisions are key to regulating the number and patterning of stomata in Arabidopsis thaliana (L.) Heynh. Many formative asymmetric divisions take place in neighbor cells (NCs), cells adjacent to a stoma or stomatal precursor. TOO MANY MOUTHS is a receptor-like protein required for the correct plane of NC division, resulting in the placement of the new precursor distal to the pre existing stoma. Because plant cells usually become polarized before asymmetric division, we studied whether NCs display a cytological asymmetry as a function of cell stage and of possible division behavior. Cells that divided in the developing leaf epidermis were smaller than 400 micro m(-2) in area and included NCs as well as isolated cells. All NCs in the youngest complexes divided with comparable frequencies, but divisions became restricted to the smaller and most recently produced NCs as the stomatal complex matured. The majority of developing NCs had distally located nuclei, suggesting that nuclear position is actively regulated in NCs. NC stages exhibiting distally located nuclei were the likeliest to divide asymmetrically. However, a distal nucleus did not necessarily predict an asymmetric division, because more NCs had distal nuclei than were likely to divide. No defect was detected in nuclear distribution in tmm NCs. These data suggest that TMM uses intercellular signals to control the plane of asymmetric division after or independently of nuclear positioning. PMID- 12569399 TI - Light strongly promotes gene transfer from Agrobacterium tumefaciens to plant cells. AB - Light conditions during Agrobacterium-based plant transformation, the most routinely used method in plant genetic engineering, differ widely and, to our knowledge, have not been studied systematically in relation to transformation efficiency. Here, light effects were examined in two already optimized transformation procedures: coculture of Agrobacterium tumefaciens with callus from two genotypes of the crop plant Phaseolus acutifolius (tepary bean) and coculture of root segments from two ecotypes of Arabidopsis thaliana. Except for the light conditions during coculture, all steps followed established procedures. Coculture was done either under continuous darkness, under a commonly used photoperiod of 16 h light/8 h darkness or under continuous light. beta glucuronidase (GUS) production due to the transient expression of an intron containing uidA gene in the binary vector was used to evaluate T-DNA transfer. In all situations, uidA expression correlated highly and positively with the light period used during coculture; it was inhibited severely by darkness and enhanced more under continuous light than under a 16 h light/8 h dark photoperiod. The promotive effect of light was observed with Agrobacterium strains harboring either a nopaline-, an octopine- or an agropine/succinamopine-type non-oncogenic helper Ti plasmid. The observed positive effect of light has obvious implications for developing and improving transient and stable transformation protocols, specifically those involving dark coculture conditions. PMID- 12569401 TI - Expression and localization of human lysozyme in the endosperm of transgenic rice. AB - In order to understand the characteristics of recombinant protein expression and sublocalization in rice ( Oryza sativa L.) endosperm, we examined the expression level of human lysozyme protein and its subcellular location in transgenic rice seeds driven by rice glutelin and globulin promoters and signal peptides. A time course of human lysozyme expression during endosperm development was analyzed. The results showed that the expression profile of recombinant protein accumulation in endosperm paralleled that of the two storage proteins. Immunofluorescence microscopy revealed that human lysozyme and storage proteins co-localized to type-II protein bodies. Both promoter-signal peptide parings targeted recombinant protein to the protein bodies. In addition, a transgenic line with a higher lysozyme expression level exhibited morphologically different protein bodies with an unbalanced composition of lysozyme and native storage proteins. The high-level expression of recombinant protein distorted the trafficking and sorting of native storage proteins in rice endosperm and affected the expression of native storage protein. PMID- 12569400 TI - MSI-99, a magainin analogue, imparts enhanced disease resistance in transgenic tobacco and banana. AB - Magainin is one of the earliest reported antimicrobial peptides isolated from skin secretions of the African clawed frog Xenopus laevis. A synthetic substitution analogue of magainin, MSI-99, is employed in this study to impart disease resistance in transgenic tobacco ( Nicotiana tabacumL.) and banana [( Musaspp. cv. Rasthali (AAB)]. This peptide inhibited the growth and spore germination of Fusarium oxysporumf.sp. cubenseat 16 micro g/ml. MSI-99 has been subcloned into plant expression vectors pMSI164 and pMSI168, targeting the peptide into the cytoplasm and extracellular spaces, respectively. Tobacco plants transformed with pMSI168 showed enhanced resistance against Sclerotinia sclerotiorum, Alternaria alternataand Botrytis cinerea. Transgenic banana pants were obtained for both pMSI164 and pMSI168 transformations and showed resistance to F. oxysporumf.sp. cubenseand Mycosphaerella musicola. The transgenic nature of the transformants and expression of this peptide was confirmed through polymerase chain reaction (PCR) and reverse transcription (RT)-PCR. The results suggest that MSI-99 can be useful in imparting enhanced disease resistance in transgenic plants. PMID- 12569402 TI - Effects of co-expressing the plant CDK inhibitor ICK1 and D-type cyclin genes on plant growth, cell size and ploidy in Arabidopsis thaliana. AB - The cyclin-dependent kinase (CDK) plays a crucial role in regulating the cell cycle of eukaryotic organisms including plants. From previous studies, it is known that ICK1, the first plant CDK inhibitor identified in Arabidopsis plants, interacts with Arath;CycD3;1 (CycD3) and Arath;CDKA;1 (Cdc2a). Overexpression of ICK1 has major effects on cell division, plant growth, and morphology. In this study, approaches were taken to determine the effects on transgenic 35S::ICK1 Arabidopsis plants of introducing another gene that could potentially modulate the activity of ICK1. F1 plants were obtained by crossing 35S::ICK1 plants with wild type (Wt) and transgenic plants expressing 35::GUS, 35S::CycD3, 35S::CycD2, or 35S::antiICK1 ( antiICK1 refers to antisense- ICK1). The major effects on plant growth and morphology observed in the 35S::ICK1 plants were partially reversed in the F1 plants from the crosses [35S::ICK1 x 35S::CycD2] and [35S::ICK1 x 35S::CycD3], and completely restored in the F1 plants from the cross [35S::ICK1 x 35S::antiICK1]. This observation was further supported by the results of ploidy analysis and structural characterization. Overexpression of CycD2 and CycD3 had the opposite effect on leaf cell size to the overexpression of ICK1. In addition, in ICK1-overexpressing plants, the CycD2 and CycD3 transcript levels increased, indicating a possible feedback regulation. The present results demonstrate that the interactions between ICK1 and D-type cyclins previously observed by the yeast two-hybrid and in vitro techniques are biologically relevant. These results illustrate the possibility of modifying plant growth and architecture dynamically by adjusting the levels of positive and negative cell-cycle regulators. PMID- 12569403 TI - Investigating the role of polyols in Cladosporium fulvum during growth under hyper-osmotic stress and in planta. AB - The role of the large amounts of polyols accumulated by the fungal tomato pathogen, Cladosporium fulvum (syn. Fulvia fulva, Cooke) both in planta and in axenic cultures has been examined. Arabinitol and glycerol accumulated in response to hyper-osmotic stress in vitro. Mannitol levels were lower in osmo stressed mycelium. (13)C NMR spectroscopy indicated that carbon flow from glucose to mannitol was redirected to arabinitol and glycerol in hyper-osmotic conditions. Infected tomato ( Lycopersicon esculentum Mill.) plants contained all three polyols whereas glycerol was the only polyol present in uninfected plants, suggesting that the mannitol and arabinitol were of fungal origin. Substantially higher levels of arabinitol and glycerol were present in infected plants that were subjected to a restricted watering regime compared to fully watered plants. The results suggest that a primary role of fungal arabinitol and glycerol, but not mannitol, is osmoregulation and that water acquisition is an important aspect of pathogenicity. PMID- 12569404 TI - Intracellular feruloylation of arabinoxylan in wheat: evidence for feruloyl glucose as precursor. AB - Incorporation of [(3)H]arabinose and [(14)C]ferulic acid into soluble and polymeric fractions from suspension-cultured wheat (Triticum aestivum L.) cells and the corresponding extracellular medium was studied. The major part of these products was identified as arabinoxylan and two proteins of 40 and 100 kDa. The time course suggests an intracellular synthesis of feruloylated arabinoxylan with feruloyl-glucose as substrate. In contrast, synthesis of feruloylated proteins appears to occur with feruloyl-CoA as precursor. Intracellular formation of ferulic acid dimers is limited to 8,5'-diferulic acid, while other dimers appear to be formed extracellularly. [(3)H]Arabinose was incorporated into polymeric material in both the cellular and in the medium fraction while [(14)C]ferulic was only found in polymers from the cellular fraction, indicating synthesis of both feruloylated and non-feruloylated arabinoxylan by the cells. PMID- 12569405 TI - Measurement of photochemical quenching of absorbed quanta in photosystem I of intact leaves using simultaneous measurements of absorbance changes at 830 nm and thermal dissipation. AB - The relationship between the redox state of the photosystem (PS) I primary donor, P700, and thermal energy dissipation in PSI were examined in intact leaves using simultaneous measurements of absorbance changes at 830 nm and variations of thermal emission monitored by photoacoustic (PA) spectroscopy, respectively. A strict proportionality (close to a 1:1 ratio) was found between the magnitudes of P700 oxidation and a positive variable PA signal induced by far-red light of various irradiances under conditions favoring effective electron donation from PSII to PSI. The proportionality was observed also between the ratio of reduced P700 to the total P700 content and the ratio of the variable component to the total PA signal measured with modulated light of 695 nm. Those findings clearly revealed that in intact leaves, variable thermal dissipation in PSI is determined by the fraction of P700 in the reduced state. Diuron-treated leaves exposed to 45 degrees C in which PSI received electrons not from PSII, but from soluble reductants localized in the chloroplast stroma were also used. In such leaves, the linear relationship between the ratio of reduced P700 to the total P700 content and the ratio of the variable component to the total PA signal measured with modulated light of 700 nm has been found as well, but its slope was twice smaller than in untreated leaves. This is probably related to an increased contribution of thermal emission from inactive PSII to the steady-state level of the PA signal in diuron-treated leaves exposed to high temperatures. The results demonstrated that the yield of variable thermal dissipation is strictly dependent on the redox pressure applied to the photosystem. The above illustrates the strong photochemical energy quenching occurring when the reaction centers are in open state (reduced P700). PMID- 12569406 TI - Glutamate synthesis in barley roots: the role of the plastidic glucose-6 phosphate dehydrogenase. AB - Evidence is provided for a close link between glutamate (Glu) synthesis and the production of reducing power by the oxidative pentose phosphate pathway (OPPP) in barley ( Hordeum vulgare L. var. Alfeo) root plastids. A rapid procedure for isolating organelles gave yields of plastids of over 30%, 60% of which were intact. The formation of Glu by intact plastids fed with glutamine and 2 oxoglutarate, both substrates of glutamate synthase (GOGAT), depends on glucose-6 phosphate (Glc-6-P) supply. The whole process exhibited an apparent K(m Glc-6-P) of 0.45 mM and is abolished by azaserine, a specific inhibitor of GOGAT; ATP caused a decrease in the rate of Glu formation. Glucose and other sugar phosphates were not as effective in supporting Glu synthesis with respect to Glc 6-P; only ribose-5-phosphate, an intermediate of OPPP, supported rates equivalent to Glc-6-P. Glucose-6-phosphate dehydrogenase (Glc6PDH) rapidly purified from root plastids showed an apparent K(m Glc-6-P) of 0.96 mM and an apparent K(m NADP)(+) of 9 micro M. The enzyme demonstrated high tolerance to NADPH, exhibiting a K(i) (NADPH) of 58.6 micro M and selectively reacted with antibodies against potato plastidic, but not chloroplastic, Glc6PDH isoform. The data support the hypothesis that plastidic OPPP is the main site of reducing power supply for GOGAT within the plastids, and suggest that the plastidic OPPP would be able to sustain Glu synthesis under high NADPH:NADP(+) ratios even if the plastidic Glc6PDH may not be functioning at its highest rates. PMID- 12569407 TI - Characterization of salt stress-enhanced phosphoenolpyruvate carboxylase kinase activity in leaves of Sorghum vulgare: independence from osmotic stress, involvement of ion toxicity and significance of dark phosphorylation. AB - C(4) phosphoenolpyruvate carboxylase (PEPCase: EC 4.1.1.31) is subjected to in vivo regulatory phosphorylation by a light up-regulated, calcium-independent protein kinase. Salt stress greatly enhanced phosphoenolpyruvate carboxylase kinase (PEPCase-k) activity in leaves of Sorghum. The increase in PEPCase-k anticipated the time course of proline accumulation thereby suggesting that water stress was not involved in the kinase response to salt. Moreover, osmotic stress seemed not to be the main factor implicated, as demonstrated by the lack of effect when water availability was restricted by mannitol. In contrast, LiCl (at a concentration of 10 mM in short-term treatment of both excised leaves and whole plants) mimicked the effects of 172 mM NaCl salt-acclimation, indicating that the rise in PEPCase-k activity resulted primarily from the ionic stress. Both NaCl and LiCl treatments increased the activity of a Ca(2+)-independent, 35 kDa kinase, as demonstrated by an in-gel phosphorylation experiment. Short-term treatment of excised leaves with NaCl or LiCl partially reproduces the effects of whole plant treatments. Finally, salinization also increased PEPCase-k activity and the phosphorylation state of PEPCase in darkened Sorghum leaves. This fact, together with increased malate production during the dark period, suggests a shift towards mixed C(4) and crassulacean acid metabolism types of photosynthesis in response to salt stress. PMID- 12569408 TI - Seed yield and plant biomass increases in rice are conferred by deregulation of endosperm ADP-glucose pyrophosphorylase. AB - In this work we test the hypothesis that yield of rice ( Oryza sativa L.) can be enhanced by increasing endosperm activity of ADP-glucose pyrophosphorylase (AGP), a key enzyme in starch biosynthesis. The potential for increases in yield exist because rice initiates more seeds than are taken to maturity and possesses excess photosynthetic capacity that could be utilized if there were more demand for assimilate. Following an approach already shown to be successful in wheat, experiments were designed to increase demand for assimilate by increasing the capacity for starch synthesis in endosperm. This was accomplished by transforming rice with a modified maize AGP large subunit sequence ( Sh2r6hs) under control of an endosperm-specific promoter. This altered subunit confers upon AGP decreased sensitivity to allosteric inhibition by inorganic phosphate (Pi) and enhanced heat stability, potentially leading to higher AGP activity in vivo. The Sh2r6hs transgene increased AGP activity in developing endosperm by 2.7-fold in the presence of Pi. Increases in AGP activity in transgenic seeds compared with controls were maximal between 10-15 days after anthesis. Starch content of individual seeds at harvest was not increased, but seed weight per plant and total plant biomass were each increased by more than 20%. Increased endosperm AGP activity thus stimulates setting of additional seeds and overall plant growth rather than increasing yield of seeds already set. Results demonstrate that deregulation of endosperm AGP increases overall plant sink strength, leading to larger, more productive plants in a manner similar to that in wheat having similar genetic modification. PMID- 12569409 TI - Quantitative relationships between induced jasmonic acid levels and volatile emission in Zea mays during Spodoptera exigua herbivory. AB - Jasmonic acid (JA) has long been hypothesized to be an important regulator of insect-induced volatile emission; however, current models are based primarily on circumstantial evidence derived from pharmacological studies. Using beet armyworm caterpillars (BAW: Spodoptera exigua) and intact corn seedlings, we examine this hypothesis by measuring both the time-course of insect-induced JA levels and the relationships between endogenous JA levels, ethylene, indole and sesquiterpenes. In separate Morning and Evening time-course trials, BAW feeding stimulated increases in JA levels within the first 4-6 h and resulted in maximal increases in JA, indole, sesquiterpenes and ethylene 8-16 h later. During BAW herbivory, increases in JA either paralleled or preceded the increases in indole, sesquiterpenes and ethylene in the Morning and Evening trials, respectively. By varying the intensity of the BAW herbivory, we demonstrate that strong positive relationships exist between the resulting variation in insect-induced JA levels and volatile emissions such as indole and the sesquiterpenes. To address potential signaling interactions between herbivore-induced JA and ethylene, plants were pretreated with 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. 1-MCP pretreatment resulted in reduced production of ethylene and volatile emission following BAW herbivory but did not alter the insect-induced accumulation of JA. Our results strongly support a role for JA in the regulation of insect-induced volatile emission but also suggest that ethylene perception regulates the magnitude of volatile emission during herbivory. PMID- 12569410 TI - Nod factors activate both heterotrimeric and monomeric G-proteins in Vigna unguiculata (L.) Walp. AB - Nod factors are lipo-chito-oligosaccharides secreted by rhizobia that initiate many responses in the root hairs of the legume hosts, culminating in deformed hairs. The heterotrimeric G-protein agonists mastoparan, Mas7, melittin, compound 48/80 and cholera toxin provoke root hair deformation, whereas the heterotrimeric G-protein antagonist pertussis toxin inhibits mastoparan and Nod factor NodNGR[S] (from Rhizobiumsp. NGR234) induced root hair deformation. Another heterotrimeric G-protein antagonist, isotetrandrine, only inhibited root hair deformation provoked by mastoparan and melittin. These results support the notion that G proteins are implicated in Nod factor signalling. To study the role of G-proteins at a biochemical level, we examined the GTP-binding profiles of root microsomal membrane fractions isolated from the nodulation competent zone of Vigna unguiculata(L.) Walp. GTP competitively bound to the microsomal membrane fractions labelled with [(35)S]GTPgammaS, yielding a two-site displacement curve with displacement constants ( K(i)) of 0.58 micro M and 0.16 mM. Competition with either ATP or GDP revealed a one-site displacement curve with K(i) of 4.4 and 29 micro M, respectively, whereas ADP and UTP were ineffective competitors. The GTP binding profiles of microsomal membrane fractions isolated from roots pretreated with either NodNGR[S] or the four-sugar, N- N'- N"- N'"-tetracetylchitotetraose (TACT) backbone of Nod factors were significantly altered compared with control microsomal fractions. To identify candidate proteins, membrane proteins were separated by SDS-PAGE and electrotransferred to nitrocellulose. GTP overlay experiments revealed that membrane fractions isolated from roots pretreated with NodNGR[S] or TACT contained two proteins (28 kDa and 25 kDa) with a higher affinity for GTPgammaS than control membrane fractions. Western analysis demonstrated that membranes from the pretreated roots contained more of another protein (~55 kDa) recognised by Galpha(common) antisera. These results provide pharmacological and biochemical evidence supporting the contention that G proteins are involved in Nod factor signalling and, importantly, implicate monomeric G-proteins in this process. PMID- 12569411 TI - Elucidation of the functions of genes central to diterpene metabolism in tobacco trichomes using posttranscriptional gene silencing. AB - The functions of two key, trichome-expressed genes were assessed using different posttranscriptional gene silencing strategies (PTGS). Efficient RNA interference (RNAi) revealed the function of a cembratriene-ol (CBT-ol) cyclase gene responsible for conversion of geranylgeranyl pyrophosphate to CBT-ols, and verified the function of a P450 gene responsible for conversion of CBT-ols to CBT diols. CBT-diols are abundant diterpenes that comprise about 60% and 10% of trichome exudate weight and leaf dry weight, respectively, in Nicotiana tabacum, T.I. 1068. The relative efficiencies and levels of suppression using antisense (AS), sense co-suppression (S), and RNAi were compared for these two genes. With a partial cDNA of the P450 gene, the suppression efficiencies (percent of primary transformants with high CBT-ols/CBT-diols) were low, 3.3% for AS and 0% for S plants. In contrast, using RNAi with a partial gene sequence, a knockdown efficiency of about 45% was achieved. For the CBT-ol cyclase gene, no suppression was observed using partial cDNAs in AS or S orientations, while RNAi with a partial gene sequence yielded an efficiency of about 64%. The efficiencies of gene silencing using full-length coding regions of both genes in AS and S orientations were 500 nm). The photocyclization quantum yields were found to be close to unity irrespective of the crystal types, while the photocycloreversion quantum yields were different as much as four times depending on the conformation of the closed-ring isomers in the crystals. PMID- 12569454 TI - Determining the strengths of hydrogen bonds in solid-state ammonia and urea: insight from periodic DFT calculations. AB - Plane-wave density functional theory has been applied to determine the strengths of hydrogen bonds in the phase I crystal structures of ammonia and urea. For ammonia, each component of the trifurcated hydrogen bond has been found to be almost as strong as a standard N-H.N interaction, and for urea the strengths of the two different N-H.O interactions have been determined by a quantum mechanical technique for the first time. PMID- 12569455 TI - Halide anion capture and recognition by a tetrahedral tetraammonium receptor in water: a molecular dynamics investigation. AB - We report molecular dynamics potential of mean force (PMF) simulations on the capture of halide anions X(-) (F(-), Cl(-), Br(-)) by a tetrahedral receptor L(4+) built from four quaternary ammonium sites connected by six (CH(2))(n) chains, leading to the formation of inclusion complexes X(-) subset L(4+). Simulations performed with a reaction field correction of the electrostatics and with PME-Ewald summation gave very similar energy profiles. In aqueous solution, an energy barrier of 12-17 kcal mol(-1) was found for the three anions, mainly due to their dehydration when they enter through the largest triangular face of L(4+). In the inclusion complexes, the anion is anchored near the center of the cavity due to the electrostatic field of the four positively charged ammonium sites, shielded from the surrounding water molecules. It was predicted that L(4+) is selective for Cl(-) over Br(-) which both form stable inclusion complexes, while the F(-) complex should dissociate. The comparison of PMFs in aqueous solution and in the gas phase and the energy component analysis demonstrates the importance of solvent on the nature of these complexes and on the complexation energy profiles. The Cl(-)/Br(-) selectivity obtained from the dissociation pathways in water was in good agreement with the results of free energy perturbation simulations based on the "alchemical route" of a thermodynamic cycle, and consistent with experimental observations. PMID- 12569456 TI - Topological rotational strengths as chirality descriptors for fullerenes. AB - A graph-theoretical procedure is proposed for assigning a chirality descriptor (the topological sign tau(+) or tau(-)) to each enantiomer of a chiral polyhedron, polyhedral molecule or graph, independently of any vertex labelling scheme. Model Cartesian coordinates and rotational strengths are obtained using only adjacency information; a generalised HOMO-LUMO rotational strength is used to associate a sign with a Schlegel diagram and the corresponding three dimensional structure, polyhedron or molecule. The topological sign gives an unambiguous way of communicating the identity of an enantiomer. The mean-square topological rotational strength is a possible measure of the chirality content of a polyhedral graph or structure. PMID- 12569457 TI - Synthesis of enantiomerically-pure [13C]aristeromycylcobalamin and its reactivity in dioldehydratase, glyceroldehydratase, ethanolamine ammonia-lyase and methylmalonyl-CoA mutase reactions. AB - We describe a novel enantioselective synthesis of aristeromycin, the carbocyclic analogue of adenosine. The seven-step synthesis is also suitable for the preparation of specifically-labelled [6'-(13)C]aristeromycin. Both the unlabelled and (13)C-labelled product was coupled to vitamin B(12) to form aristeromycylcobalamin. This carbocyclic analogue of coenzyme B(12) was examined for its coenzymic activity with several adenosylcobalamin-dependent enzymes. For glyceroldehydratase and dioldehydratase, the reaction rate (k(cat)) was 38 and 44 % of that measured with adenosylcobalamin as coenzyme. In contrast, aristeromycylcobalamin showed no detectable activity with methylmalonyl-CoA mutase and ethanolamine ammonia-lyase. Instead, it was a weak inhibitor of the former and a strong inhibitor of the latter enzyme. The slower turnover rate with glyceroldehydratase raised the hope of detecting the 6'-deoxyaristeromycyl radical intermediate. Comparison of the EPR spectra of the intermediates in the glyceroldehydratase reaction, which used adenosyl- and aristeromycylcobalamines, respectively, as coenzyme, revealed a significant shift and this suggests a different geometric position of these cofactors at the binding site during the cleavage of the carbon-cobalt bond. However, we found no evidence for the existence of a 6'-deoxyaristeromycyl radical during the reaction with [6' (13)C]aristeromycylcobalamin. We conclude that the lifetime of this radical is still too short to be observed. PMID- 12569458 TI - Redox-induced coordination isomerization of a phosphoniobenzophospholide. AB - 1-Triphenylphosphoniobenzo[c]phospholide 1 reacts with [M(CO)(5)Br] (M = Mn, Re) and [Mn(CO)(3)(naphthalene)][BF(4)] to give complexes cis-[M(CO)(4)(1)Br] (5 a,b) and [Mn(CO)(3)(1)][BF(4)] (6 a[BF(4)]), respectively, featuring eta(1)(P)- and eta(5)(pi)-coordination of the phosphole ring. The corresponding reactions with [M(2)(CO)(10)] proceed with conservation of the metal-metal bond and yield, depending on the reaction temperature, dinuclear complexes [M(2)(CO)(8)(1)] (M=Mn, 7 a) or [M(2)(CO)(6)(1)(2)] (M=Mn, Re, 8 a,b) with mu(2)-bridging eta(1)(P):eta(2)(Pdbond;C) coordination of the phosphole moiety. All complexes formed were characterized by spectroscopic data; 5 b, 6 a[BF(4)], and 8 a,b were characterized by X-ray diffraction studies as well. The structural and (31)P NMR data of the dinuclear manganese complex 8 a suggest that the interaction between the metal atoms and the eta(2)-bound Pdbond;C double bond moieties is dominated by the L-->M charge-transfer contribution; this hints at a very low back-donation ability of the central M(2)(CO)(6) fragment. Investigation of the reactions of the Mn complexes 6 a and 8 a with Mg or ferrocenium hexafluorophosphate ([Fc][PF(6)]), respectively, revealed that the chemically reversible mutual interconversion between both species was feasible. Likewise, oxidation of the rhenium complex 8 b with [Fc][PF(6)] gave spectroscopic evidence for the formation of a Re analogue of 6 a. Electrochemical studies suggested that the oxidation 8 a-->2 6 a involves two consecutive single-electron-transfer steps, the first of which is electrochemically reversible and produces a metastable radical cation that is detectable by ESR spectroscopy. The mutual interconversion between 6 a and 8 a represents the first case of a reversible coordination isomerization of a phosphaarene that is triggered by a redox process and might stimulate further studies directed at the use of dinuclear phosphaarene complexes in redox-catalysis. PMID- 12569459 TI - Early-late heterobimetallic alkoxides as model systems for late-transition-metal catalysts supported on titania. AB - Titanium complexes with chelating alkoxide ligands [TiCp*(O(2)Bz)(OBzOH)] (1) and [TiCp*(Me)((OCH(2))(2)Py)] (2) were synthesised by reaction of [TiCp*Me(3)] (Cp*=eta(5)-C(5)Me(5)) with 2-hydroxybenzyl alcohol ((HO)(2)Bz) and 2,6 pyridinedimethanol ((HOCH(2))(2)Py), respectively. Complex 1 reacts with [(M(mu OH)(cod))(2)] (M=Rh, Ir) to yield the early-late heterobimetallic complexes [TiCp*(O(2)Bz)(2)M(cod)] [M=Rh (3), Ir (4)]. Carbon monoxide readily replaces the COD ligand in 3 to give the rhodium dicarbonyl derivative [TiCp*(O(2)Bz)(2)Rh(CO)(2)] (5). Compound 2 reacts with [(M(mu-OH)(cod))(2)] (M=Rh, Ir) with protonolysis of a Tibond;Me bond to give [TiCp*((OCH(2))(2)Py)(mu O)M(cod)] [M=Rh (6), Ir (7)]. The molecular structures of complexes 3, 5 and 7 were established by single-crystal X-ray diffraction studies. PMID- 12569460 TI - New mechanistic insight into the coupling reactions of CO2 and epoxides in the presence of zinc complexes. AB - Coupling reactions of CO(2) and epoxide to produce cyclic carbonates were performed in the presence of a catalyst [L(2)ZnX(2)] (L=pyridine or substituted pyridine; X=Cl, Br, I), and the effects of pyridine and halide ligands on the catalytic activity were investigated. The catalysts with electron-donating substituents on pyridine ligands exhibit higher activity than those with unsubstituted pyridine ligands. On the other hand, the catalysts with electron withdrawing substituents at the 2-position of the pyridine ligands show no activity; this demonstrates the importance of the basicity of the pyridine ligands. The catalytic activity of [L(2)ZnX(2)] was found to decrease with increasing electronegativity of the halide ligands. A series of highly active zinc complexes bridged by pyridinium alkoxy ions of the general formula [((mu OCHRCH(2)L)ZnBr(2))(n)] (n=2 for R=CH(3); n=3 for R=H; L=pyridine or substituted pyridine) were synthesized and characterized by X-ray crystallography. The dinuclear zinc complexes obtained from propylene oxide adopt a square-planar geometry for the Zn(2)O(2) core with two bridging pyridinium propoxy ion ligands. Trinuclear zinc complexes prepared from ethylene oxide adopt a boat geometry for the Zn(3)O(3) core, in which three zinc and three oxygen atoms are arranged in an alternate fashion. These zinc complexes bridged by pyridinium alkoxy ions were also isolated from the coupling reactions of CO(2) and epoxides performed in the presence of [L(2)ZnBr(2)]. Rapid CO(2) insertion into the zincbond;oxygen bond of the zinc complexes bridged by pyridinium alkoxy ions leads to the formation of zinc carbonate species; these which yield cyclic carbonates and zinc complexes bridged by pyridinium alkoxy ions upon interaction with epoxides. The mechanistic pathways for the formation of active species and cyclic carbonates are discussed on the basis of results from structural and spectroscopic analyses. PMID- 12569461 TI - Synthesis and structures of trifluoromethyl-, fluoro-, and azido-substituted hexabenzylhexaazaisowurtzitanes and isolation of a novel hexaazaisowurtzitane based polycycle. AB - The first hexabenzylhexaazaisowurtzitane cage compounds with trifluoromethyl and azide groups, as well as those with fluorine atoms in the 3- and 4-positions, have been prepared and fully characterized. A study of the substituent influence on the benzene ring regarding the formation of the hexaazaisowurtzitane polycycle in either the 2- or 4-position with CF(3), F, and N(3) revealed an interesting difference. In all cases with CF(3), F, and N(3) substituents in the 4-position, the corresponding hexabenzylhexaazaisowurtzitanes were isolated. The corresponding hexabenzylhexaazaisowurtzitanes were also formed when these substituents were in the 2-position; however, in addition with azide in 2 position a novel type of polycycle was isolated and identified. (15)N NMR data and crystal structures of hexabenzylhexaazaisowurtzitane derivatives were obtained and are discussed in detail. PMID- 12569462 TI - A new class of ferromagnetically-coupled mixed valence vanadium(IV/V) polyoxometalates. AB - Reaction of [V(VI)OCl(2)(thf)(2)] with a bidentate nitrogen-donor ligand (L: phen=1,10-phenanthroline, 5-mephen=5-methyl-1,10-phenanthroline, bipy=2,2' bipyridine, 5,5'-me(2)bipy=5,5'-dimethyl-2,2'-bipy) in methyl alcohol, in the presence of triethylamine, leads to the formation of hexameric [V(2) (IV)V(4) (V)] oxo-alkoxo-vanadates of the general formula [V(6)O(12)(mu(2) OCH(3))(4)(L)(4)].x H(2)O [L=phen (1.4 H(2)O), 5-mephen (2.6 H(2)O), bipy (3.4 H(2)O), 5,5'-me(2)bipy (4.H(2)O)]. X-ray structure analysis of 1.2 H(2)O and 4.8 CH(3)OH revealed a pair of V(3)O(13)N(4) trimeric units sharing two corners, with a centrosymmetric planar V(6)-core. In addition, a fully oxidized V(V) species [V(V) (4)O(8)(OCH(3))(2)(mu(3)-OCH(3))(2)(5,5'-me(2)bipy)(2)].3 CH(3)OH (5.3 CH(3)OH) was isolated from the reaction mixture used for the synthesis of 4.H(2)O. The crystal structure of 5.3 CH(3)OH revealed a dicubane-like framework with two missing vertices. Electron paramagnetic resonance (EPR) and variable temperature magnetic susceptibility studies for the hexamers 1.4 H(2)O and 3.4 H(2)O showed the complete localization of the single 3d electrons on the V(IV) ions and unusual ferromagnetic interaction between the two paramagnetic vanadium(IV) ions separated by a distance of about 5.1 A. Furthermore, intermolecular antiferromagnetic interactions through pi-contacts of phenyl rings were observed for these species below 8 K. The ferromagnetic exchange coupling observed in the hexanuclear compounds 1.4 H(2)O and 3.4 H(2)O is also discussed using ab initio UHF calculations on a model compound. The value of the exchange coupling constant (3.7 cm(-1)) for this model compound, calculated using the broken symmetry approach, is in good agreement, both in sign and magnitude, with the experimental J values (6.00 cm(-1) for 1.4 H(2)O and 8.54 cm(-1) for 3.4 H(2)O). PMID- 12569463 TI - Template synthesis of benzannulated N-heterocyclic carbene ligands. AB - The reaction of 2-azidophenyl isocyanide (7) with [M(CO)(5)(thf)] (M=Cr, W) yields the isocyanide complexes [M(CO)(5)(7)] (M=Cr 8, M=W 9). Complexes 8 and 9 react with tertiary phosphines such as triphenylphosphane at the azido function of the isocyanide ligand to give the 2-triphenylphosphiniminophenyl isocyanide complexes 10 (M=Cr) and 11 (M=W). The polar triphenylphosphiniminophenyl function in complexes 10 and 11 can be hydrolyzed with H(2)O/HBr to afford triphenylphosphane oxide and the complexes containing the unstable 2-aminophenyl isocyanide ligand. This ligand spontaneously cyclizes by intramolecular nucleophilic attack of the primary amine at the isocyanide carbon atom to yield the 2,3-dihydro-1H-benzimidazol-2-ylidene complexes 12 (M=Cr) and 13 (M=W). Double deprotonation of the cyclic NH,NH-carbene ligands in 12 and 13 with KOtBu and reaction with two equivalents of allyl bromide yields the N,N'-dialkylated benzannulated N-heterocyclic carbene complexes 14 (M=Cr) and 15 (M=W). The molecular structures of complexes 9 and 11-15 were confirmed by X-ray diffraction studies. PMID- 12569464 TI - Competitive reactions of interstrand and intrastrand DNA-Pt adducts: A dinuclear platinum complex preferentially forms a 1,4-interstrand cross-link rather than a 1,2 intrastrand cross-link on binding to a GG 14-mer duplex. AB - A study of the kinetics and mechanism of the reaction between the dinuclear Pt complex [(trans-PtCl(NH(3))(2))(2)(mu-NH(2)(CH(2))(6)NH(2))](2+) (1) and the 14 mer duplex 5'-d(ATACATG(7)G(8)TACATA)-3'.5'-d(TATG(25)TACCATG(18)TAT)-3' is reported. [(1)H,(15)N]-HSQC NMR was used to follow the reaction at 298 K, pH 5.4. The product is primarily the 5'-5' 1,4-interstrand cross-link between G(8) and G(18) bases and exists in two conformational forms. No evidence for the possible 1,2-intrastrand G(7)G(8) adduct was seen, confirming the preferential formation of interstrand cross-links by these dinuclear complexes. An initial electrostatic association of (15)N-1 with the duplex is indicated by changes in its (1)H/(15)N chemical shifts, followed by aquation of 1 to form the monoaqua monochloro species 2, with a rate constant of 4.00+/-0.03x10(-5) s(-1). Monofunctional binding to the duplex occurs primarily at G(8), the 3' base of the nucleophilic GG grouping, with a rate constant of 1.5+/-0.7 M(-1) s(-1). Changes in the (1)H/(15)N shifts indicate there is an electrostatic interaction between the unbound (PtN(3)Cl) group of the monofunctional adduct and the duplex. No peaks for a transient aquated monofunctional species are seen and closure of 3 to form the 1,4-G(8)G(18) interstrand cross-link (5) was treated as direct, with a rate constant of 4.47+/-0.06x10(-5) s(-1). The G(8)G(18) cross-link was confirmed from analysis of the NOESY NMR spectrum of the final product. Structural perturbations for the 1,4-interstrand cross-link extend over approximately four base-pairs and are similar to those found for a 1,4-interstrand cross-link with a shorter 8-mer GTAC- sequence. A major distortion was evident for the 5'T (T(17)) adjacent to the platinated G(18), consistent with the findings from the use of chemical probes to investigate the conformation of 1,4-interstrand cross-links. PMID- 12569465 TI - An ab initio/RRKM study of product branching ratios in the photodissociation of buta-1,2- and -1,3-dienes and but-2-yne at 193 nm. AB - Ab initio G2M(MP2)//B3LYP/6-311G** calculations have been performed to investigate the reaction mechanism of photodissociation of buta-1,2- and -1,3 dienes and but-2-yne after their internal conversion into the vibrationally hot ground electronic state. The detailed study of the potential-energy surface was followed by microcanonical RRKM calculations of energy-dependent rate constants for individual reaction steps (at 193 nm photoexcitation and under collision-free conditions) and by solution of kinetic equations aimed at predicting the product branching ratios. For buta-1,2-diene, the major dissociation channels are found to be the single Cbond;C bond cleavage to form the methyl and propargyl radicals and loss of hydrogen atoms from various positions to produce the but-2-yn-1-yl (p1), buta-1,2-dien-4-yl (p2), and but-1-yn-3-yl (p3) isomers of C(4)H(5). The calculated branching ratio of the CH(3) + C(3)H(3)/C(4)H(5) + H products, 87.9:5.9, is in a good agreement with the recent experimental value of 96:4 (ref. 21) taking into account that a significant amount of the C(4)H(5) product undergoes secondary dissociation to C(4)H(4) + H. The isomerization of buta-1,2 diene to buta-1,3-diene or but-2-yne appears to be slower than its one-step decomposition and plays only a minor role. On the other hand, the buta-1,3-diene- >buta-1,2-diene, buta-1,3-diene-->but-2-yne, and buta-1,3-diene-->cyclobutene rearrangements are significant in the dissociation of buta-1,3-diene, which is shown to be a more complex process. The major reaction products are still CH(3) + C(3)H(3), formed after the isomerization of buta-1,3-diene to buta-1,2-diene, but the contribution of the other radical channels, C(4)H(5) + H and C(2)H(3) + C(2)H(3), as well as two molecular channels, C(2)H(2) + C(2)H(4) and C(4)H(4) + H(2), significantly increases. The overall calculated C(4)H(5) + H/CH(3) + C(3)H(3)/C(2)H(3) + C(2)H(3)/C(4)H(4) + H(2)/C(2)H(2) + C(2)H(4) branching ratio is 24.0:49.6:4.6:6.1:15.2, which agrees with the experimental value of 20:50:8:2:2022 within 5 % margins. For but-2-yne, the one-step decomposition pathways, which include mostly H atom loss to produce p1 and, to a minor extent, molecular hydrogen elimination to yield methylethynylcarbene, play an approximately even role with that of the channels that involve the isomerization of but-2-yne to buta-1,2- or -1,3-dienes. p1 + H are the most important reaction products, with a branching ratio of 56.6 %, followed by CH(3) + C(3)H(3) (23.8 %). The overall C(4)H(5) + H/CH(3) + C(3)H(3)/C(2)H(3) + C(2)H(3)/C(4)H(4) + H(2)/C(2)H(2) + C(2)H(4) branching ratio is predicted as 62.0:23.8:2.5:5.7:5.6. Contrary to buta-1,2- and -1,3-dienes, photodissociation of but-2-yne is expected to produce more hydrogen atoms than methyl radicals. The isomerization mechanisms between various isomers of the C(4)H(6) molecule including buta-1,2- and -1,3 dienes, but-2-yne, 1-methylcyclopropene, dimethylvinylidene, and cyclobutene have been also characterized in detail. PMID- 12569466 TI - Preorganized bis-zinc phosphodiester cleavage catalysts possessing natural ligands: a lesson pertinent to bimetallic artificial enzymes. AB - Two preorganized bis-zinc receptors (2 and 3) were synthesized wherein the metals were ligated with ligands present in natural phosphodiesterases: imidazoles and carboxylates. The intrametallic distance is near 4.5 A, that found in natural nucleases and other successful artificial nucleases. With only two imidazoles (2), the zinc binding affinities were not high enough to achieve cooperativity. Yet, with a third ligand, a carboxylate (3), cooperativity was found in the cleavage of HPNPP. The preorganization of 3 was achieved using a "steric gearing" strategy. The enhancement was 80-fold for cooperation between the two metals relative to a mono-metallic analogue (5). However, there was no observable enhancement in the hydrolysis of RNA using 3 relative to 5. Therefore, we conclude that placing two zinc atoms that are ligated with natural ligands at the appropriate distance for catalysis is not sufficient to enhance the cleavage of RNA, but is successful for activated RNA substrate mimics. PMID- 12569467 TI - An alternative way of characterising the bonding in compounds featuring main group elements and with the potential for multiple bonding: on the dissociation of binary main-group hydrides. AB - Herein the bonding in compounds featuring main-group elements and with the potential for multiple bonding is studied theoretically by examination of their fragmentation into two fragments that still exhibit the same structure as they had in the molecule prior to dissociation. The fragments were calculated both in their electronic ground state and in an excited electronic state, in which the number of unpaired electrons is equal to the maximal number of bonds in the compounds before dissociation. The energies of the fragmentation processes (DeltaE(frag)) can be more directly linked to the bond strengths than the dissociation energies (DeltaE(diss)), because of the absence of any secondary effects like relaxation of the electronic state or of the geometry of the fragments. These relaxation energies of the fragments (DeltaE(frag)) are also studied herein. The energies derived in this work allow for an accurate comparison of the bonding properties in main-group-element hydrides. The trends of the fragmentation and relaxation energies are discussed in detail. It will be shown that the relaxation energies allow for a classification of the bonds ("classical" sigma and pi bonds or donor-acceptor interactions), while the fragmentation energies are good quantitative measures for the total bond strength. Similar calculations are on the way to explore the bonding in systems in which the hydrogen atoms are replaced by organic groups or halogen atoms. PMID- 12569468 TI - Prevalence of the alkyl/phenyl-folded conformation in benzylic compounds C6H5CH2 X-R (X=O, CH2, CO, S, SO, SO2): significance of the CH/pi interaction as evidenced by high-level ab initio MO calculations. AB - Ab initio MO calculations were carried out to examine the conformational energies of various benzylic compounds C(6)H(5)CH(2)XR (X=O, CH(2), CO, S, SO, SO(2); R=CH(3), C(2)H(5), iC(3)H(7), tC(4)H(9)) at the MP2/6-311G(d,p)//MP2/6-31G(d) level. Rotamers with R/Ph in gauche relationship are generally more stable than the R/Ph anti rotamers. In these stable geometries, the interatomic distance in the interaction of alpha- or beta-CH in the alkyl group and the ipso-carbon atom of the phenyl ring is short. The computational results are consistent with experimental data from supersonic molecular jet spectroscopy on 3-n-propyltoluene and NMR and crystallographic data on structurally related ketones, sulfoxides, and sulfones. In view of this, the alkyl/phenyl-congested conformation of these compounds has been suggested to be a general phenomenon, rather than an exception. The attractive CH/pi interaction has been suggested to be a dominant factor in determining the conformation of simple aralkyl compounds. PMID- 12569469 TI - Dynamic chemical devices: generation of reversible extension/contraction molecular motion by ion-triggered single/double helix interconversion. AB - The polyheterocyclic strands 1-H and 2-H adopt a helical shape enforced by the pyridine-pyrimidine helicity codon. The crystal structure of 2-H shows the formation of stacks of dimers of right- and left-handed individual helices. Treatment of 1-H and 2-H with silver triflate results in the generation of double helical entities 1-DH and 2-DH, containing two strands and two silver ions. NMR studies and determination of the crystal structure of 2-DH indicate that the duplex is stabilized by coordination of each Ag(+) ion to two terminal bipyridine units, one from each strand, and by pronounced pi-pi stacking interactions between the internal heterocycles of the strands, yielding a very robust double helical structure. Reversible interconversion of the single and double helix may be achieved by addition of a cryptand capable of sequestering Ag(+) and releasing it by protonation. Thus, successive addition of acid and base leads to reversible interconversion between the shorter ( approximately 3.6 A) single helix and the longer ( approximately 10.3 A) double helix, resulting in the generation of pronounced extension/contraction motion. The system 1,2-H/1,2-DH represents a dynamic chemical device undergoing ionic modulation of reversible molecular mechanical motion fueled by acid/base neutralization. PMID- 12569470 TI - Synthesis and solid-state NMR studies of p-vinylbenzylphosphonic acid. AB - P-Vinylbenzylphosphonic acid (2) was synthesized and crystallized in two phases A and B. The latter phase was easily converted into the former. Both phases were investigated by IR and solid-state (31)P and (1)H NMR spectroscopy, and X-ray crystallography. The use of recently developed NMR methods has enabled us to increase the usually poor resolution of solid-state (1)H spectra. This gave additional insights on the proton environments. In particular, two dimensional 2D (1)H-(31)P heteronuclear correlation (HETCOR) experiments, incorporating a Lee Golburg homonuclear decoupling scheme, allowed description of the mixture of phase A and B. PMID- 12569471 TI - Highly enantioselective conjugate addition of AlMe3 to linear aliphatic enones by a designed catalyst. AB - 2-Hydroxy-2'-alkylthio-1,1'-binaphthyl compounds are catalytic promoters of the 1,4-addition of AlMe(3) to linear aliphatic enones in THF at -40 to -48 degrees C in the presence of [Cu(MeCN)(4)]BF(4). At ligand loadings of 5-20 mol %, enantioselectivities of 80-93 % are realised for most substrates. To attain these values, the use of highly pure AlMe(3) is mandatory. The presence of methylalumoxane (MAO), derived by hydrolysis, leads to reduced enantioselectivity and a conjugate addition product. PMID- 12569472 TI - Recognition of caffeine in aqueous solutions. AB - Binding of caffeine in aqueous solutions has been achieved for the first time by using water-soluble, tetracationic peptide-porphyrin conjugates Zn-1 as the receptor molecules. The association constant for caffeine with receptor Zn-1 is in some cases as high as 6000 M(-1), only 5-6 times lower than the highest binding constant reported for an artificial caffeine receptor in low polarity aprotic solvents. The binding mechanism has been studied by a combination of experimental techniques such as UV-visible and NMR spectroscopy and microcalorimetry. Recognition of caffeine involves both stacking with the porphyrin ring and metal coordination. Subtle variations of the receptor structure affect the complexation. Receptors Zn-1 have also been investigated for the recognition of molecules structurally related to caffeine, for example, 1 methylimidazole. Selectivity towards oxopurine derivatives (caffeine and theophylline) have been found. PMID- 12569473 TI - Unidirectional threading of triphenylureidocalix[6]arene-based wheels: oriented pseudorotaxane synthesis. AB - Triphenylureidocalix[6]arenes 5 a,b are heteroditopic receptors having a pinched cone structure able to interact with both the cation and the anion of ion pairs. They are able to act as wheels and form complexes of the pseudorotaxane type with axles derived from dialkylviologen salts. An investigation into the possibility of exploiting the different structural and chemical information present on the two distinct rims of the calixarene wheel as control elements to pivot the direction of the axle threading processes and give access to oriented pseudorotaxanes is reported. It was verified that, in C(6)D(6), an asymmetric dicationic axle derived from 4,4'-bipyridil bearing two alkyl chains, one of which has a stopper, and triphenylureidocalix[6]arenes 5 a or 5 b form 1:1 supramolecular complexes belonging to the class of pseudorotaxanes. The structure of these complexes has been inferred through (1)H NMR techniques. The data show that the axle accesses the calixarene cavity only through the wider rim. To further verify this issue, the new rotaxane 8, obtained by stoppering the pseudorotaxane derived from 5 b and the symmetrical axle 7 with diphenylacetyl chloride, was synthesised. In the (1)H NMR spectrum of 8, the aliphatic protons of the axle portion that resides at the wide rim of the wheel show chemical shifts that are almost identical to those observed in pseudorotaxanes 6. On the other hand, those that stick out of the narrow rim of 8 experience chemical shifts that could not be found in the oriented pseudorotaxanes 6. PMID- 12569474 TI - Molecular switch triggered by solvent polarity: synthesis, Acid-base behavior, alkali metal ion complexation, and crystal structure. AB - The synthesis and characterization of the new tetraazamacrocycle L, bearing two 1,1'-bis(2-phenol) groups as side-arms, is reported. The basicity behavior and the binding properties of L toward alkali metal ions were determined by means of potentiometric measurements in ethanol/water 50:50 (v/v) solution (298.1+/-0.1 K, I=0.15 mol dm(-3)). The anionic H(-1)L(-) species can be obtained in strong alkaline solution, indicating that not all of the acidic protons of L can be removed under the experimental conditions used. This species behaves as a tetraprotic base (log K(1)=11.22, log K(2)=9.45, log K(3)=7.07, log K(4)=5.08), and binds alkali metal ions to form neutral [MH(-1)L] complexes with the following stability constants: log K(Li)=3.92, log K(Na)=3.54, log K(K)=3.29, log K(Cs)=3.53. The arrangement of the acidic protons in the H(-1)L(-) species depends on the polarity of the solvents used, and at least one proton switches from the amine moiety to the aromatic part upon decreasing the polarity of the solvent. In this way two different binding areas, modulated by the polarity of solvents, are possible in L. One area is preferred by alkali metal ions in polar solvents, the second one is preferred in solvents with low polarity. Thus, the metal ion can switch from one location to the other in the ligand, modulated by the polarity of the environment. A strong hydrogen-bonding network should preorganize the ligand for coordination, as confirmed by MD simulations. The crystal structure of the [Na(H(-1)L)].CH(3)CN complex (space group P2(1)/c, a=12.805(1), b=20.205(3), c=14.170(2) A, beta=100.77(1) degrees, V=3601.6(8) A(3), Z=4, R=0.0430, wR2=0.1181), obtained using CH(2)Cl(2)/CH(3)CN as mixed solvent, supports this last aspect and shows one of the proposed binding areas. PMID- 12569478 TI - Enzymatic catalysis of the peptidyl-prolyl bond rotation: are transition state formation and enzyme dynamics directly linked? PMID- 12569479 TI - Recent developments in protein microarray technology. AB - The sequencing of the human genome and the advent of DNA chips and sophisticated bioinformatics platforms have enabled molecular biologists to take a more global view of biological systems and to analyze naturally occurring genetic variation. Microarrays of antibodies can measure the concentrations of many proteins quickly and simultaneously. Microarrays of genomically encoded proteins allow scientists to screen entire genomes for proteins that interact with particular factors, catalyze particular reactions, or act as substrates for protein-modifying enzymes or as targets of autoimmune responses. The new protein microarray platforms will prove invaluable to basic biological research, and will dramatically accelerate the pace of discovery of drug targets and diagnostic biomarkers. PMID- 12569480 TI - One century of aryne chemistry. AB - Arynes, which are formally derived from aromatic rings by abstraction of two hydrogen atoms, have been a focus of organic chemistry for 100 years. In contrast to ortho-benzyne, which is mentioned in almost every introductory textbook on organic reaction mechanisms as a reactive intermediate of nucleophilic aromatic substitution, the meta and para isomers were regarded as rather exotic until recently. This situation has changed dramatically with the discovery of the enediyne antibiotics, a promising new class of antitumor drugs, and has aroused the interest of research groups from all branches of chemistry. Nowadays, arynes and related compounds are among the most intensively studied systems in chemistry. However, many aspects of the chemistry of these reactive intermediates are not well understood yet. In this review we outline the historical developement with an emphasis on recent progress in this challenging field of research. PMID- 12569481 TI - A hexanuclear iridium chain. PMID- 12569482 TI - Nanoporous lanthanide-copper(II) coordination polymers: syntheses and crystal structures of [[M2(Cu3(iminodiacetate)6)].8 H2O]n (M=La, Nd, Eu). PMID- 12569484 TI - Stabilization of tricoordinate pyramidal boron: theoretical studies on CBSiH5, BSi2H5, CBGeH5, and CBSnH5. PMID- 12569483 TI - Design and application of an alpha-helix-mimetic scaffold based on an oligoamide foldamer strategy: antagonism of the Bak BH3/Bcl-xL complex. PMID- 12569485 TI - RPM-1: a recyclable nanoporous material suitable for ship-in-bottle synthesis and large hydrocarbon sorption. PMID- 12569486 TI - The preparation of a double metallahelicate containing 28 copper atoms. PMID- 12569487 TI - Total synthesis of (+/-)-photodeoxytridachione through a Lewis acid catalyzed cyclization. PMID- 12569489 TI - Exact replication of biological structures by chemical vapor deposition of silica. PMID- 12569488 TI - [[N(PPh3)2]2[Os18Pd3(mu6-C)2(CO)42]]: an osmium-palladium mixed-metal high nuclearity carbonyl cluster. PMID- 12569490 TI - Surface-initiated polymerization on self-assembled monolayers: amplification of patterns on the micrometer and nanometer scale. PMID- 12569492 TI - [2.2]Paracyclophanes with defined substitution pattern-key compounds for the mechanistic understanding of the Gilch reaction to poly(p-phenylene vinylene)s. PMID- 12569491 TI - Coordinated o-dithio- and o-iminothiobenzosemiquinonate(1-) pi radicals in [MII(bpy)(L*)](PF6) complexes. PMID- 12569494 TI - DIBAH-mediated amide/hydride transformation in ansa-lanthanidocene(III) complexes. PMID- 12569493 TI - The surprising structures of B8F12 and B10F12. PMID- 12569495 TI - An efficient and selective photocatalytic system for the oxidation of sulfides to sulfoxides. PMID- 12569496 TI - Highly stereoselective N-terminal functionalization of small peptides by chiral phase-transfer catalysis. PMID- 12569500 TI - The 50th birthday of the organocuprates. PMID- 12569501 TI - Advances in functional-group-tolerant metal-catalyzed alkyl-alkyl cross-coupling reactions. PMID- 12569502 TI - Bioactivation of nitroglycerin--a new piece in the puzzle. PMID- 12569503 TI - Sensory principles of higher plants. AB - Plants respond to a broad variety of stimuli from the interior of their body and from the outside environment. A revolution in our knowledge of the sensory capabilities of plants took place during the last decade, thanks to the consequent application of the tools of molecular genetics and the focusing of most work, in a world-wide effort, on a single, representative, higher plant: Arabidopsis thaliana. This review concentrates on the complete sensory periphery of higher plants, with focus placed on the principles rather than the details, and such systems that have been functionally identified unambiguously. Higher plants are no less fascinating, no less complex, and no less sensitive than animals, even man, with respect to their sensory capabilities. Plants constantly monitor their environment and their interior situation by using a stunning array of sensory systems, that are mostly different from those of animals or man. PMID- 12569504 TI - Hierarchical and self-similar growth of self-assembled crystals. PMID- 12569506 TI - A cyclic metallopeptide induces alpha helicity in short peptide fragments of thermolysin. PMID- 12569505 TI - Sliding helix and change of coordination geometry in a model di-MnII protein. PMID- 12569507 TI - Polyoxovanadium(IV) sulfite compounds: synthesis, structural, and physical studies. PMID- 12569508 TI - Porous coordination-polymer crystals with gated channels specific for supercritical gases. PMID- 12569509 TI - Toward fully synthetic N-linked glycoproteins. PMID- 12569510 TI - A strategy for the synthesis of phosphorus-containing macrocycles--ligands for exceptional coordination geometries. PMID- 12569511 TI - Application of HRMAS 1H NMR spectroscopy to investigate interactions between ligands and synthetic receptors. PMID- 12569512 TI - A topologically chiral molecular dumbbell. PMID- 12569513 TI - Triple bonding to tin: synthesis and characterization of the stannylyne complex trans-[Cl(PMe3)4W[triple bond]Sn-C6H3-2,6-Mes2]. PMID- 12569514 TI - Iodine-induced gold-nanoparticle fusion/fragmentation/aggregation and iodine linked nanostructured assemblies on a glass substrate. PMID- 12569515 TI - Dendritic stars by ring-opening-metathesis polymerization from ruthenium-carbene initiators. PMID- 12569516 TI - Large, concentration-dependent enhancement of the quadratic hyperpolarizability of [Zn(CH3CO2)2(L)2] in CHCl3 on substitution of acetate by triflate. PMID- 12569517 TI - Dual stereoselectivity of 1-(2'-carboxy)benzyl 2-deoxyglycosides as glycosyl donors in the direct construction of 2-deoxyglycosyl linkages. PMID- 12569518 TI - Porphyrazines as molecular scaffolds: periphery-core spin coupling between metal ions of a schiff base porphyrazine. PMID- 12569519 TI - Copper and silver complexes containing the S(SiMe2S)2(2-) ligand: efficient entries into heterometallic sulfido clusters. PMID- 12569523 TI - Background and operating parameters in microemulsion electrokinetic chromatography. AB - Microemulsion electrokinetic chromatography (MEEKC) is an electrodriven separation technique. Separations are generally achieved using microemulsions consisting of surfactant-coated nanometer-sized oil droplets suspended in aqueous buffer. A cosurfactant such as a short-chain alcohol is generally used to stabilize the microemulsion. This review summarizes the various microemulsion types and compositions that have been used in MEEKC. The effects of key-operating variables such as surfactant type and concentration, cosurfactant type and concentration, buffer pH and type, oil type and concentration, use of organic solvent and cyclodextrin additions, and temperature are described. Specific examples of water-in-oil microemulsions and chirally selective separations are also covered. PMID- 12569524 TI - Retention modeling and resolution optimization for a group of N-phenylpyrazole derivatives in micellar electrokinetic chromatography using empirical and physicochemical models. AB - The optimization of the separation resolution for a group of N-phenylpyrazole derivatives in micellar electrokinetic chromatography (MEKC) as a function of the separation buffer composition (surfactant and organic modifier concentration) has been performed. In order to achieve our purpose, the first step has been the prediction of the migration times of the electroosmotic flow (t(0)) and micelles (t(m)), and the retention factors of solutes (k), as a function of surfactant (sodium dodecyl sulfate) and alcohol (n-propanol or n-butanol) concentrations, by means of empirical equations. Also, some physicochemical models have been applied to relate the retention factors to the surfactant and the organic modifier concentrations in order to optimize the separation resolution and to increase our knowledge of the separation process. Finally, a comparison of the resolution optimization through the use of the physicochemical and empirical models selected has been made in order to obtain the optimum separation buffer composition for the separation of a group of 17 N-phenylpyrazole derivatives as test solutes. PMID- 12569525 TI - Analysis of selected withanolides in plant extract by capillary electrochromatography and microemulsion electrokinetic chromatography. AB - Microemulsion electrokinetic chromatography (MEEKC) coupled with a diode-array detector was developed for the simultaneous analysis of natural steroidal compounds, withanolides including withaferin A, withacnistin and iochromolide. Optimal resolution was obtained with a microemulsion consisting of 70 mM octane, 800 mM 1-butanol, 100 mM sodium dodecyl sulfate (SDS), and 10 mM phosphate-borate buffer (pH 7) using a fused-silica capillary at 25 kV and 40 degrees C. Since this technique is not compatible with mass spectrometry detection, a capillary electrochromatographic method was developed to separate the investigated withanolides. The effects of mobile phase composition and pH were systematically investigated. Complete separation was obtained with a capillary electrochromatography (CEC) Hypersil C18 bonded silica column (packed length, 25 cmx100 microm ID and 375 microm OD), packed with 3 microm particles. The mobile phase consisted of formic acid-ammonia, pH 8 / acetonitrile (40/60 v/v); the voltage was set at 25 kV and the temperature at 20 degrees C. Under these conditions, resolution of these closely related compounds, including the critical pair withacnistin and iochromolide, was achieved in less than 5 min. The separations by MEEKC and CEC were compared with that obtained by reversed-phase liquid chromatography and showed similar retention order, indicating the analogy of the retention mechanism of these techniques. To further improve specificity and sensitivity, the developed CEC method was interfaced with electrospray ionization mass spectrometry using a Teflon connection between the CEC column and a void fused-silica capillary. Finally, the described methods were applied to the qualitative analysis of withanolides in Iochroma gesnerioides plant extract. PMID- 12569526 TI - Enhancement of selectivity and resolution in the enantioseparation of uncharged compounds using mixtures of oppositely charged cyclodextrins in capillary electrophoresis. AB - The enantiomeric separation of some nonsteroidal anti-inflammatory drugs (NSAIDs) was investigated in capillary electrophoresis (CE) using dual systems with mixtures of charged cyclodextrin (CD) derivatives. A significant enhancement of selectivity and resolution could be achieved in the enantioseparation of these analytes in their uncharged form by the simultaneous addition of two oppositely charged CD derivatives to the background electrolyte. The combination of the single-isomer cationic CD, permethyl-6-monoamino-6-monodeoxy-beta-CD (PMMAbetaCD) and the single-isomer polyanionic CD, heptakis-6-sulfato-beta-cyclodextrin (HSbetaCD) in a pH 2.5 phosphoric acid-triethanolamine buffer, was designed and employed for the enantioseparation of profens. The improvement in selectivity and resolution can be attributed to the fact that the two CDs, which lead to independent and enantioselective complexation with the analyte enantiomers, have not only opposite effects on the electrophoretic mobility of these compounds but also opposite affinity patterns towards the enantiomers of these compounds. Binding constants for these enantiomers with each CD were determined using linear regression approach, in order to be able to predict the effect of the concentrations of the two CDs on enantiomeric selectivity and resolution in such dual systems. PMID- 12569527 TI - Synthesis, analytical characterization and use of octakis(2,3-di-O-methyl-6-O sulfo)-gamma-cyclodextrin, a novel, single-isomer, chiral resolving agent in low pH background electrolytes. AB - The third member of the family of single-isomer, sulfated gamma-cyclodextrins, the sodium salt of octakis(2,3-di-O-methyl-6-O-sulfo)-gamma-cyclodextrin has been synthesized, analytically characterized and used for the capillary electrophoretic separation of the enantiomers of nonionic, weak acid and weak base analytes in low-pH aqueous background electrolytes. Though octakis(2,3-di-O methyl-6-O-sulfo)-gamma-cyclodextrin complexes less strongly with many of the analytes tested than the other members of the single-isomer, 6-O-sulfo gamma cyclodextrin family, such as octa(6-O-sulfo)-gamma-cyclodextrin and octakis(2,3 di-O-acetyl-6-O-sulfo)-gamma-cyclodextrin, it offers excellent separation selectivities, often complementary to those of both the single-isomer, 6-O-sulfo beta-cyclodextrins and 6-O-sulfo gamma-cyclodextrins. Rapid, efficient enantiomer separations were observed for a large number of structurally diverse analytes in acidic aqueous background electrolytes. PMID- 12569528 TI - Synergistic effects of ion-pairing in the enantiomeric separation of basic compounds with cyclodextrin derivatives in nonaqueous capillary electrophoresis. AB - The enantiomeric separation of various kinds of basic pharmaceuticals has been investigated in nonaqueous capillary electrophoresis (NACE) systems using an ion pairing reagent in combination with cyclodextrins (CDs). The simultaneous addition to the methanolic background electrolyte (BGE) of (+)-S-camphorsulfonate or alkanesulfonates and an anionic beta-cyclodextrin derivative, heptakis(2,3 dimethyl-6-sulfato)-beta-cyclodextrin (HDMS-beta-CD), led to partial or complete enantioresolution in most cases. In the absence of ion-pairing reagent, the enantiomeric resolution obtained with this CD derivative was most often completely lost or strongly reduced, indicating the important role of ion-pairing in the chiral recognition mechanism in these NACE systems. The influence of the nature and concentration of the counterion and the anionic CD derivative on the enantioseparation of basic compounds was studied. Synergistic effects between these two kinds of charged additives were clearly observed. PMID- 12569529 TI - Influence of methanol as a buffer additive on the mobilities of organic cations in capillary electrophoresis. AB - The mobilities of a series of aromatic ammonium ions, ranging in charge from +1 to + 3, were investigated by capillary electrophoresis using buffers consisting of 0-75% v/v methanol. This is an extension of our previous studies involving anion mobility in methanol-water media [1]. Absolute mobilities were determined by extrapolation of the effective mobilities to zero ionic strength according to the Pitts' equation. For all of the buffer compositions studied, the ionic strength effect increased with increasing cation charge, and varied as a function of solvent 1/eta epsilon (1/2) as predicted by the electrophoretic term within the Pitts' equation. In the presence of methanol, the ionic strength effects became more dramatic. The absolute mobilities of the cations were altered by the addition of methanol to the electrophoretic media. For example, at 75% MeOH, a migration order reversal was observed between the + 2 and + 3 ammonium ions. These solvent-induced selectivity changes are attributed to dielectric friction. As predicted by the Hubbard-Onsager dielectric friction model, dielectric friction increased with increasing methanol content and with increasing analyte charge. Further, the changes in cation mobility correlated to the changes in solvent relaxation time (tau), epsilon and eta. Although not predicted by the Hubbard-Onsager theory, the + 3 ammonium ion experienced more dielectric friction than the - 3 sulfonate and - 3 carboxylate investigated previously [1]. This apparent failure of the Hubbard-Onsager model results from its continuum nature, whereby ion-solvent interactions are not taken into account. PMID- 12569530 TI - Mathematical modelling of adsorption and transport processes in capillary electrochromatography: open-tubular geometry. AB - A mathematical modelling approach for open-tubular capillary electrochromatography is presented. The spatially one-dimensional model takes into account (i) a coupling of (non)linear adsorption of positively or negatively charged analyte molecules (at a negatively charged capillary inner surface) with the equilibrium electrokinetics at this solid-liquid interface, (ii) mobile phase transport by electroosmosis and pressure-driven flow, as well as (iii) transport of species by electrophoresis and molecular diffusion. Under these conditions the local zeta-potential and electroosmotic mobility become a function of the concentration of the charged analyte. The resulting inhomogeneity of electroosmotic flow through the capillary produces a compensating pore pressure as requirement for incompressible mobile phase flow (i.e., for constant volumetric flow along the capillary). The results of the simulations are discussed in view of the surface-to-volume ratio of the capillary lumen, the analyte concentration (in combination with a Langmuir isotherm for the adsorption process), and buffer effects. PMID- 12569531 TI - Estimation and comparison of zeta-potentials of silica-based anion-exchange type porous particles for capillary electrochromatography from electrophoretic and electroosmotic mobility. AB - Mobilities of different chromatographic particles obtained from two electrokinetic methods were determined and compared. The particles were all based on porous silica, between 3 and 15 microm diameter, and were either native, or derivatized. As intermediate of chemical modification 3-mercaptopropyl-modified silica particles (TP-silica) are obtained. These particles were finally transformed into weakly basic anion exchangers with O-9-(tert butylcarbamoyl)quinine (tBuCQN) as chiral selector. The electrophoretic mobility of the particles was determined from their migration velocity in an electric field using microelectrophoresis. Electrokinetic chromatography with a capillary column packed with the same particles was used to measure the electroosmotic flow generated. All measurements were carried out in background electrolytes of equal ionic strength (10(-2) mol/L), at pH varying between 3.5 and 9.5. From these data a rough estimation of the zeta-potential was made, taking Helmholtz-Smoluchowski conditions into consideration. With both methods the zeta-potential of the native silica particles is negative throughout, and its value increases with pH. The weakly basic tBuCQN particles have positive zeta-potentials at pH lower than about 7.5, but exhibit a negative zeta-potential above this pH, indicating the dominating effect of residual silanol groups at the silica surface. The zeta potential for these anion-exchange particles ranged between +30 and -40 mV. The zeta-potentials derived with electrophoresis and electroosmosis agree, showing the adequacy of the approach, although many limitations must be taken into account in the treatment of the electrokinetic phenomena in such porous systems. These restrictions in interpreting mobility and zeta-potential were discussed. PMID- 12569532 TI - Control of electroosmotic flow in nonaqueous capillary electrophoresis by polymer capillary coatings. AB - In aqueous capillary electrophoresis the electroosmotic flow (EOF) can be strongly suppressed or eliminated by coating the capillary surface silanols either by buffer additive adsorption or chemical modification. Hydrophilic coatings, e.g., polyvinyl alcohol (PVA) proved to be most efficient for EOF control in applications like DNA analysis. In nonaqueous capillary electrophoresis (NACE), however, the EOF cannot be totally suppressed with these capillaries and coating efficiency turned out to be solvent-depending. In this paper, fused-silica capillaries with monomeric and polymeric coatings differing in hydrophobicity and chemical properties (vinyl, vinyl acetate, vinyl alcohol and acrylates with different alkyl chain length) were investigated. Besides studying the EOF characteristics with different organic solvents and water, gas chromatography (GC) measurements were carried out to probe the silanol reduction via ether retention and the surface hydrophobicity by retention of nonane. Good correlations between GC results and EOF magnitude could be found. It could be demonstrated that the polymeric coating has to be solvatized by the buffer solvent to reduce the EOF. The PVA coating was optimal for aqueous systems but not effective for some nonaqueous buffers. On the other hand, polyvinyl acetate and polyethyl acrylate as polymeric coatings proved to be optimal to reduce the EOF in NACE. PMID- 12569533 TI - Capillary electrochromatography with monolithic silica column: I. Preparation of silica monoliths having surface-bound octadecyl moieties and their chromatographic characterization and applications to the separation of neutral and charged species. AB - Monolithic silica columns with surface-bound octadecyl (C18) moieties have been prepared by a sol-gel process in 100 microm ID fused-silica capillaries for reversed-phase capillary electrochromatography of neutral and charged species. The reaction conditions for the preparation of the C18-silica monoliths were optimized for maximum surface coverage with octadecyl moieties in order to maximize retention and selectivity toward neutral and charged solutes with a sufficiently strong electroosmotic flow (> 2 mm/s) to yield rapid analysis time. Furthermore, the effect of the pore-tailoring process on the silica monoliths was performed over a wide range of treatment time with 0.010 M ammonium hydroxide solution in order to determine the optimum time and conditions that yield mesopores of narrow pore size distribution that result in high separation efficiency. Under optimum column fabrication conditions and optimum mobile phase composition and flow velocity, the average separation efficiency reached 160 000 plates/m, a value comparable to that obtained on columns packed with 3 microm C18 silica particles with the advantages of high permeability and virtually no bubble formation. The optimized monolithic C18-silica columns were evaluated for their retention properties toward neutral and charged analytes over a wide range of mobile phase compositions. A series of dimensionless retention parameters were evaluated and correlated to solute polarity and electromigration property. A dimensionless mobility modulus was introduced to describe charged solute migration and interaction behavior with the monolithic C18-silica in a counterflow regime during capillary electrochromatography (CEC )separations. The mobility moduli correlated well with the solute hydrophobic character and its charge-to-mass ratio. PMID- 12569534 TI - Sieving mechanisms in polymeric matrices. AB - A critical review of the existing theoretical models and experimental evidences for sieving mechanisms during separation of macromolecules, paying particular attention to capillary electrophoresis applications is presented. Gel models (Ogston and reptation) have been successfully applied to highly entangled polymer solutions, where fast and efficient separations can occur. In order to account for the DNA/polymers collision-interaction mechanisms during separation in dilute solutions - characterized by a poorer resolution -, approximated analytical models have been developed. An insight in the mechanism regulating the intermediate case of moderately entangled polymer solutions, for low fields and concentrations of small multiples of the overlap concentration c*, is given by the constraint release approach. This model proposes an upper limit of size separation, increasing with matrix concentration and molecular mass. Finally, the coupling between the reptative motion of the analytes and the effect of matrix constraint release very likely plays a fundamental role in the separation mechanism and requires therefore further and deeper investigation, both theoretically and experimentally. PMID- 12569535 TI - An exactly solvable Ogston model of gel electrophoresis: X. Application to high field separation techniques. AB - Recently, we generalized our lattice model of gel electrophoresis to study the net velocity of particles being pulled by a high-intensity electric field through an arbitrary distribution of immobile obstacles (Gauthier, M. G., Slater, G. W., J. Chem. Phys. 2002, 117, 6745-6756). In this article, we show how the high-field version of our model can be used to compare the velocity of particles with different electric charges and/or physical sizes. We then investigate specific two-dimensional distributions of obstacles that can be used to separate particles, e.g., in a microfluidic device. More precisely, we compare the velocity of differently charged or sized analytes in sieving, trapping and deflecting systems to model various electrophoretic separation techniques. In particular, we study the nonlinear effects present in ratchet systems and how they can be combined with time-asymmetric pulsed fields to provide new modes of separation. PMID- 12569536 TI - Anomalously fast migration of triplet-repeat DNA in capillary electrophoresis with linear polymer solution. AB - We investigated the capillary electrophoretic behavior of single-stranded DNA fragments in methylcellulose solution, and found that triplet-repeat DNA showed anomalously faster mobilities than DNA markers with random sequence. Through the further study on the electrophoretic data, reptation model is proven appropriate to describe the migration of DNA under our experimental conditions. Accordingly, with the equations based on reptation theory, we could obtain the persistence length of DNA fragments and find that these values of triplet-repeat DNAs are larger than that of DNA markers with random sequence, which means the former DNAs are less flexible than the latter ones when they migrate in the electric field. This phenomenon is supposed to result from the characteristic higher-order structure formed by GC base pairs within triplet-repeat DNA, which is further proven by the resumed migration order in accordance with DNA size when the denaturant is added into the sieving matrix. PMID- 12569537 TI - Integrated on-capillary instrumentation for gene expression measurement directly from cells. AB - We studied the fundamental instrumental issues relevant to a capillary-based integrated system to measure expression of a specific gene directly from cells. Samples were introduced into a capillary by use of a syringe pump. All reactions were carried out in a microthermocycler, where a part of the capillary having 1 microL inner volume was used as a reaction vessel. First, cells were lysed by heating to release RNA, followed by deoxyribonuclease (DNase) treatment. Then, reverse transcription-polymerase chain reaction (RT-PCR) was performed to obtain amplified products from the targeted mRNA. Finally, the product was verified by capillary electrophoresis (CE) with laser-induced fluorescence detection. The whole protocol was completed in the system in 3 h. PCR product from beta-actin mRNA in 16 human lymphoblast cells was obtained with a signal-to-noise ratio (S/N) of 3400 +/- 730 (n = 3). Therefore, the system is reproducible and sensitive enough to measure gene expression from a single cell. We show that the amplified fragment from breast cancer-specific mRNA was obtained from cells of breast cancer cell line, but was not obtained from cells of hepatoma cell line. These results therefore lay the foundations for future CE or microchip instrumentation for high-throughput automated gene-expression analysis. PMID- 12569538 TI - Stacking phenomena in electromigration: From basic principles to practical procedures. AB - The review describes principles of procedures and techniques used in capillary zone electrophoresis for enhancement of sensitivity that are based on increasing analyte mass in its zone during the electromigration process, for which the term stacking is generally used. Attention is paid to intrinsic stacking in samples with low conductivity, transient isotachophoretic stacking applied in samples with high conductivity, and sweeping in micellar electrokinetic chromatography. Principles of these stacking schemes are explained, new procedures and instrumental arrangements are discussed, and all contributions involving stacking principles that have been published since the year 2000 are surveyed. PMID- 12569539 TI - Sample stacking revisited: a personal perspective. AB - One of the major challenges in capillary electrophoresis and other miniaturization separation techniques is to maintain high detection sensitivity in the increasingly smaller dimension. Numerous on-column sample preconcentrating procedures, based either on electrokinetic focusing or chromatographic effects, have been developed. This review will discuss some practical approaches to sample stacking from a personal perspective. Several recent developments in sample stacking on microfluidic devices are reviewed. PMID- 12569540 TI - High-sensitive capillary zone electrophoresis analysis by electrokinetic injection with transient isotachophoretic preconcentration: electrokinetic supercharging. AB - The principle of an on-line preconcentration method for capillary zone electrophoresis (CZE) named electrokinetic supercharging (EKS), is described and based on computer simulation the preconcentration behavior of the method is discussed. EKS is an electrokinetic injection method with transient isotachophoretic process, is a powerful preconcentration technique for the analysis of dilute samples. After filling the separation capillary with supporting electrolyte, an appropriate amount of a leading electrolyte was filled and the electrokinetic injection was started. After a while, terminating electrolyte was filled subsequently and migration current was applied. This procedure enabled the introduction of a large amount of sample components from a dilute sample without deteriorating separation. Computer simulation of the electrokinetic injection revealed that EKS was effective for the preconcentration of analytes with wide mobility ranges by proper choice of transient isotachophoresis (ITP) system and electroosmotic flow (EOF) should be suppressed to increase injectable amount of analytes under constant voltage mode. A test mixture of rare-earth chlorides was used to demonstrate the uses of EKS-CZE. When a 100 microL sample was used, the low limit of detectable concentration was 0.3 microg/L (1.8 nM for Er), which was comparable or even better than that of ion chromatography and inductively coupled plasma-atomic emission spectrometry (ICP AES). PMID- 12569541 TI - Chloride present in biological samples as a tool for enhancement of sensitivity in capillary zone electrophoretic analysis of anionic trace analytes. AB - Effects originating from the variability of the sample matrix can be efficiently eliminated when the separation conditions are selected so that compounds of like charge with high concentration referred to as macrocomponents are embodied into the system of transient isotachophoresis. For stacking and separation of anionic trace analytes in biological samples, the presence of chloride is shown to be important to balance out effects of other macrocomponents that act against isotachophoretic stacking. Having acetoacetate, malate, citrate, and some drug metabolites in untreated human serum samples, the stacking mechanism of these compounds in an electrolyte system comprising 5 mM mandelic acid and epsilon aminocaproic acid, pH 3.8, is explained. Analytes are monitored by indirect UV absorption detection. Attention is paid to the minimum chloride concentration required with respect to the concentration ratio of phosphate (stacker) and lactate (destacker) present in the sample so as to ensure both stacking and separation of trace analytes. Insight into the separation process is given both with computer simulations and experiments. For selected analytes, the effect of chloride concentration on quantitative evaluation, sensitivity and limit of detection is demonstrated as well. Moreover, the applicability of the mobility window between phosphate and lactate for an additional group of metabolites is sketched. PMID- 12569542 TI - The preparation of background electrolytes in capillary zone electrophoresis: golden rules and pitfalls. AB - In this article the methodology of the design of suitable background electrolytes (BGEs) in capillary zone electrophoresis (CZE) is described. The principal aspects of the role of a BGE in CZE are discussed with respect to an appropiate migration behavior of analytes, including the transport of the electric current, the buffering of pH, the Joule heat, the electro-endosmotic flow (EOF) and the principal migration and detection modes. The impact of the composition of the BGE upon migration and detection is discussed. It is shown that the total concentration of the BGE is a principal factor and the adjustment of migrating analyte zones according to the Kohlrausch regulating function (KRF) is the principal effect in most of the sample stacking techniques. The number of co-ions and their properties are of key importance for peak shapes of the analyte peaks and for the existence of system zones. The detection of UV-transparent analytes may advanteously be done in the indirect UV mode, by using UV-absorbing co-ions, however, both peaks and dips may be expected in the UV trace in case of multiple co-ionic BGEs. Properties of BGEs can be predicted applying mathematical models and it is shown that with SystCharts, predictions can be given concerning the existence of system zones, detection modes and the peak shapes of analytes for a given BGE. Practical examples of methodological considerations are given in the design of suitable BGEs for four principal combinations of migration and detection modes. The properties of the BGEs selected are exemplified with experimental results. Golden rules are summarized for the preparation of suitable BGEs in CZE. PMID- 12569543 TI - Eigenmobilities in background electrolytes for capillary zone electrophoresis: II. Eigenpeaks in univalent weak electrolytes. AB - We analyze in detail a mathematical model of capillary zone electrophoresis (CZE) based on the conception of eigenmobilities, which are eigenvalues of the matrix tied to the linearized continuity equations. Our model considers CZE systems, where constituents are weak electrolytes and where pH of the background electrolyte may reach the full range from 0 to 14. Both hydrogen and hydroxide ions are taken into account in relations for conductivity and electroneutrality. An electrophoretic system with N constituents has N eigenmobilities. We reveal that two of the eigenmobilities have a special meaning as they exist due to the presence of hydrogen ions and hydroxide ions (in water solutions). These two eigenmobilities are responsible for the existence of two corresponding system zones (system peaks). We show that the stationary zone (injection zone, water zone, gap, peak, dip) is in many common background electrolytes composed of these two eigenzones which overlap, due to their very low electrophoretic mobility, into one zone. Other eigenmobilities give rise to system zones originating due to a possible existence of double (or multiple) coconstituents in the background electrolyte. The last group of eigenmobilities is connected with the movement of eigenzones accompanying analytes and enabling their indirect UV or conductivity detection. The model allows assessing experimentally available quantities such as effective mobility of the analyte, molar conductivity detection response, transfer ratio, and relative velocity slope and gives a picture about migration of analytes, their electromigration dispersion and signals obtained in detectors. It allows computer simulation of electropherograms and enables optimization of background electrolytes. PMID- 12569544 TI - Ampholytes as background electrolytes in capillary zone electrophoresis: sense or nonsense? Histidine as a model ampholyte. AB - A lot of phenomena, occuring in capillary zone electrophoresis (CZE), are linked with the ionic concentration of the background electrolyte (BGE). If weak bases and acids are used as BGEs in CZE, at a pH where they are scarcely ionized, the ionic concentration of the BGE is very low and this brings a strong peak broadening, limited sample stacking and low sample load. Because the electromigration dispersion increases extremely, moreover, the existence of low conductivity BGEs in CZE is a contradiction in terms. The behavior of ampholytes as BGE in CZE is examined, by means of histidine as a model ampholyte. For BGEs consisting of histidine, important parameters, including the ionic concentrations, buffer capacity, transfer ratio, and the indicator for electromigration dispersion E(1)m(1)/E(2)m(2), are calculated at various pH. Although the transfer ratio is fairly constant over the whole pH traject, the ionic concentration and buffer capacity decrease whereas the electromigration dispersion strongly increases near the pI of histidine. I.e., that ampholytes can be applied as BGEs in CZE, however, just not at pH near their pI value, except as the difference between the pK values of the basic and acidic group, the deltapK value, is very small. For ampholytes with a low deltapK value or at high concentrations, all the before-mentioned effects are less fatal, but in that case we can not speak of a real low-conductivity BGE. If ampholytes are used at pH near their pK values, the use of ampholytes as BGE is not advantageously compared with simple weak bases and acids. This has been confirmed by calculations and experiments. PMID- 12569545 TI - Sensitive indirect photometric detection of inorganic and small organic anions by capillary electrophoresis using Orange G as a probe ion. AB - This study addresses the two major problems in the use of dyes as highly absorbing probes for indirect photometric detection in capillary electrophoresis (CE). First, effective electroosmotic flow (EOF) modification or suppression to allow separation and detection of a wide mobility range of analytes is not straightforward when electrolytes containing increased dye concentrations are used. The suppression of EOF to less than + 5x10(-9) m(2)V(-1)s(-1) was achieved with a combination of a poly(ethylenimine) (PEI)-coated capillary and the addition of the neutral polymer hydroxypropylmethylcellulose (HPMC) to the background electrolyte. Second, the deterioration of baselines due to adsorption of the dye probe to the capillary wall is generally a problem. In this work, baseline quality at higher probe concentrations was significantly improved by a rather unusual but highly effective combination of a simultaneous application of a slight overpressure (25 mbar) at the injection end during the separation, and the use of a relatively narrow capillary of 50 microm inner diameter. Both measures would appear to be counterproductive. Optimisation of the probe concentration with regard to signal-to-noise ratio resulted in an electrolyte of 4 mM Orange G, 0.05% HPMC buffered at pH 7.7 by the addition of 10.0 mM histidine isoelectric buffer. Very high separation efficiencies of 128 000-297 000 plates were made possible by the relatively high probe concentration. Combined with excellent detection sensitivity, even with the introduction of hydrodynamic flow and a reduced optical path length, these measures resulted in limits of detection ranging from 0.216 to 0.912 microM with a deuterium lamp light source (248 nm) and from 0.147 to 0.834 microM with a 476 nm blue light-emitting diode (LED) light source. Reproducibility over 30 consecutive runs without changing the electrolyte was excellent, with relative standard deviation (RSD) values of 0.14 0.80% for migration time, 1.27-3.36% for peak area and 0.88-5.12% for peak heights. The optimised electrolyte was used for the analysis of inorganic anions in air filter samples, providing good agreement with results obtained by ion chromatography. PMID- 12569546 TI - Indirect laser-induced fluorescence detection for capillary electrophoresis using a frequency-doubled diode laser. AB - A blue (452 nm) frequency-doubled diode laser with a quasi-cw optical output power of 10 microW is used for indirect laser-induced fluorescence detection in combination with the capillary electrophoretic separation of inorganic anions. As fluorescing probe ion the anion of 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) was selected having an absorption maximum of 454 nm in alkaline medium. Employing a capillary coated with linear acrylamide, baseline separation of eight inorganic anions was possible within 5 min. With a separation buffer containing 50 micromol.L(-1) HPTS and 10 mmol.L(-1) lysine the limits of detection for sulfate, nitrite, nitrate, azide, thiocyanate, and chlorate were between 0.9 and 4.7 micromol.L(-1). Separation of chloride and sulfate was achieved by adding 0.25 mmol.L(-1) calcium hydroxide to the separation buffer. Inorganic anions in several mineral and tap water samples have been determined with the technique developed and results are compared to data obtained by ion chromatography in combination with conductivity detection after conductivity suppression. PMID- 12569548 TI - Decreased DNA repair gene expression among individuals exposed to arsenic in United States drinking water. AB - Arsenic is well established as a human carcinogen, but its precise mechanism of action remains unknown. Arsenic does not directly damage DNA, but may act as a carcinogen through inhibition of DNA repair mechanisms, leading indirectly to increased mutations from other DNA damaging agents. The molecular mechanism underlying arsenic inhibition of nucleotide excision repair after UV irradiation (Hartwig et al., Carcinogenesis 1997;18:399-405) is unknown, but could be due to decreased expression of critical genes involved in nucleotide excision repair of damaged DNA. This hypothesis was tested by analyzing expression of repair genes and arsenic exposure in a subset of 16 individuals enrolled in a population based case-control study investigating arsenic exposure and cancer risk in New Hampshire. Toenail arsenic levels were inversely correlated with expression of critical members of the nucleotide excision repair complex, ERCC1 (r(2) = 0.82, p < 0.0001), XPF (r(2) = 0.56, p < 0.002), and XPB (r(2) = 0.75, p < 0.0001). The internal dose marker, toenail arsenic level, was more strongly associated with changes in expression of these genes than drinking water arsenic concentration. Our findings, based on human exposure to arsenic in a US population, show an association between biomarkers of arsenic exposure and expression of DNA repair genes. Although our findings need verification in a larger study group, they are consistent with the hypothesis that inhibition of DNA repair capacity is a potential mechanism for the co-carcinogenic activity of arsenic. PMID- 12569549 TI - A cyclooxygenase-2 inhibitor, nimesulide, inhibits postinitiation phase of N nitrosobis(2-oxopropyl)amine-induced pancreatic carcinogenesis in hamsters. AB - The modification effects of nimesulide, a cyclooxygenase (COX)-2 inhibitor, administration during the postinitiation phase of pancreatic carcinogenesis were investigated in hamsters treated with N-nitrosobis(2-oxopropyl)amine (BOP). Male Syrian hamsters were given 4 weekly s.c. injections of BOP at a dose of 10 mg/kg and thereafter administered 0, 100 or 400 ppm nimesulide in the diet for 36 weeks. Additional groups of hamsters were fed 400 ppm nimesulide without prior BOP initiation or nontreated. At week 40, all surviving animals were killed and development of neoplastic and preneoplastic lesions was assessed histopathologically. The incidence of pancreatic adenocarcinomas was significantly (p < 0.05) decreased in the BOP/400 ppm nimesulide group compared to the BOP alone group. The multiplicity of total lesions of pancreatic adenocarcinoma plus atypical hyperplasia was also significantly (p < 0.05) lowered. Immunohistochemically, COX-2 was clearly expressed in pancreatic and lung tumor cells, whereas expression was not remarkably affected by the 400 ppm nimesulide treatment. Proliferating cell nuclear antigen labeling indices of pancreatic ducts were significantly (p < 0.01) reduced by nimesulide. The incidence and multiplicity of neoplastic lesions in other organs did not significantly differ among the BOP-treated groups, though only the multiplicity of lung tumors showed a tendency to decrease. No neoplastic lesions were detected in animals receiving nimesulide alone. Our results clearly indicate that nimesulide protects against BOP-induced pancreatic tumors in hamsters. PMID- 12569550 TI - Reactive oxygen species-mediated regulation of eNOS and iNOS expression in multicellular prostate tumor spheroids. AB - Nitric oxide (NO) generated by either endothelial nitric oxide synthase (eNOS) or inducible nitric oxide synthase (iNOS) may be involved in prostate tumorigenesis through the inhibition of reactive oxygen species (ROS)-induced apoptosis. Multicellular DU-145 prostate tumor spheroids endogenously generated NO that paralleled the production of ROS. With increasing spheroid size, eNOS expression was downregulated, whereas an upregulation of iNOS expression was observed. In parallel, NO generation declined, as evaluated by the NO indicator diaminofluorescein-2 diacetate (DAF-2DA), suggesting that NO generation in DU-145 tumor spheroids is mainly mediated by eNOS. Elevation of ROS by treatment of tumor spheroids with either buthionine sulfoximine (BSO) or hydrogen peroxide resulted in upregulation of eNOS, whereas iNOS was downregulated. Furthermore, eNOS expression was increased by epidermal growth factor (EGF) in a redox sensitive manner. Upregulation of eNOS after treatment with hydrogen peroxide was apparently transduced through receptor tyrosine kinase signaling pathways since it was abolished by the protein kinase C (PKC) inhibitor bisindolylmaleimide-1 (BIM-1), the p21(ras) inhibitor S-trans-trans-farnesylthiosalicylic acid (FTS), the c-Raf inhibitor ZM 336372 and PD98059, which inhibits ERK1/2 activation. Endogenous NO may serve to escape from oxidative stress-induced apoptosis since treatment of tumor spheroids with the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5 tetramethyl imidazoline-1-oxyl 3-oxide (carboxy-PTIO) as well as the NO synthase inhibitor N-omega-amino-L-arginine (L-NAA) increased cleaved caspase-3. Consequently, lowering intracellular NO levels with either L-NAA or PTIO significantly raised ROS levels, indicating that endogenously generated NO may play a role as a ROS scavenger, thereby protecting exponentially growing tumor spheroids from ROS-induced apoptosis. PMID- 12569551 TI - The product of the candidate prostate cancer susceptibility gene ELAC2 interacts with the gamma-tubulin complex. AB - ELAC2 is a novel candidate cancer susceptibility gene located on chromosome 17p: Carriers of mutations in ELAC2 display a higher risk of developing prostate cancer. Overexpression of ELAC2 in tumor cells causes a delay in G2-M progression characterized by accumulation of cyclin B levels. Consistent with a function in mitosis, further biochemical analysis revealed that ELAC2 physically interacts with the gamma-tubulin complex. This is the first biologic insight into the function of this new putative cancer susceptibility gene, providing clues of how perturbation of ELAC2 might promote tumorigenesis through irregular cell division. PMID- 12569552 TI - Suppression of invasion and peritoneal carcinomatosis of ovarian cancer cell line by overexpression of bikunin. AB - Bikunin (bik), a Kunitz-type protease inhibitor, also known as urinary trypsin inhibitor, is proposed as a main participant in the inhibition of tumor cell invasion and metastasis, possibly through the direct inhibition of cell associated plasmin activity and suppression of urokinase-type plasminogen activator (uPA) mRNA expression. In the present study, we transfected the human ovarian carcinoma cell line HRA, highly invasive cells, with an expression vector harboring a cDNA encoding for human bik. Our study was designed to investigate the effect of bik overexpression and changes in tumor cell phenotype and invasiveness in the stably transfected clones. Bik gene transfection of HRA gave the following results: 1) transfection of HRA with the bik cDNA resulted in 5 variants stably expressing functional bik; 2) bik(+) clones exhibited a significantly reduced uPA mRNA expression as compared to the parental cells; 3) bikunin negatively regulates the ERK1/2 activity; 4) secretion-blocking treatments of bik(+) clones abrogated bik-mediated suppression of ERK1/2 activation and uPA expression; 5) the regulation of invasion seen in the HRA cells is mainly mediated by the uPA-plasmin-MMP-2 system; 6) transfection of HRA with the bik gene significantly reduced invasion, but not proliferation, adhesion, or migration relative to the parental cells; and 7) animals with bik(+) clones induced reduced peritoneal dissemination and long term survival. We conclude that transfection of HRA cells with the bik cDNA constitutively suppresses ERK1/2 activation, which results in inhibition of uPA expression and subsequently reduces dissemination of bik(+) clones. PMID- 12569553 TI - Structural analysis of the ErbB-2 receptor using monoclonal antibodies: Implications for receptor signalling. AB - The extracellular part of ErbB-2 is formed by 4 domains, specifically, L1, L2 that adopt a beta-helical structure and S1, S2 that consist of several cysteine rich, EGF-fold modules. These ectodomains mediate ErbB-2 dimerisation with itself or with other members of the epidermal growth factor receptor (EGFR) family, events essential to both ErbB-2 signaling and the development of certain malignancies. The anti-ErbB-2 monoclonal antibodies N12, N28 and L87 bind to the polypeptides C531-A586, T216-C235 and C220-C235 respectively. In this study, glycine walking and random mutagenesis were used to further delineate the critical residues involved in antibody binding. A molecular model of ErbB-2 ectodomains was then constructed based on the recently published coordinates of the EGFR (EGFR) model. This model rationalized successfully many features of our epitope mapping, including their location in modules within the S1 and S2 domains and the importance of Arg545, Gln548 and Leu561 for N12 binding. Further investigation of the functional effects of the anti-ErbB-2 monoclonal antibodies demonstrated that N28 strongly stimulated ErbB-2 phosphorylation and MAPK activation whereas N12 had no effect. As bivalency is required for the action of these antibodies we propose that at least 2 different kinds of ErbB-2 homodimers can be formed as relative rotational isomers and that the S1 and S2 domains are instrumental in determining the relative orientations of the ErbB-2 homodimers, such that different signaling effects are induced. PMID- 12569554 TI - CYP enzyme polymorphisms and susceptibility to HCV-related chronic liver disease and liver cancer. AB - Cancer risk can be influenced by the exposure to endogenous or environmental toxins. Polymorphic enzymes involved in the metabolic activation/detoxification of carcinogens may account for individual variations of risk. We studied the polymorphisms of five enzymes of the P450 superfamily, CYP1A1, CYP1A2, CYP2D6, CYP2E1 and CY3A4, as risk factors for liver disease progression and cancer in hepatitis C virus-infected patients. CYP genotyping was performed by polymerase chain reaction (PCR) restriction fragment length polymorphism or allele-specific PCR. Different stages of disease were considered, as follows: 90 asymptomatic carriers and 87 chronic hepatitis, 92 cirrhosis and 91 hepatocellular carcinoma (HCC) cases. Reference allele frequencies were obtained from 99 blood donors. Allele distributions among categories were compared using the chi(2) test. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to express relative risks. Independent associations were modeled by correspondence analysis and logistic regression. Frequencies of the CYP1A1 highly inducible alleles, MspI m2 and Val, were increased in liver disease patients compared with carriers; no specific association with HCC was found. The high-activity CYP2E1 c2 allele was underrepresented among HCC patients with respect to other HCV categories, including cirrhosis. CYP2D6 poor metabolizer (PM) genotypes were significantly more frequent in healthy subjects (7.1%) and carriers (11.1%) than in hepatitis/cirrhosis (4.6%) and HCC (1.2%) patients. This was confirmed by multivariable analysis. PM genotypes protected against progressive disease as ORs reduced proportionally to stage. The age at diagnosis for HCC was anticipated in non-PM individuals. No differences were seen for CYP1A2 and CYP3A4 genes. Polymorphic variants of CYP genes may contribute to the progression of liver disease and HCC risk in HCV-infected subjects. PMID- 12569555 TI - Frequent monoallelic deletion of PTEN and its reciprocal associatioin with PIK3CA amplification in gastric carcinoma. AB - Mutational alterations of PTEN and PIK3CA, which negatively and positively regulate PI3-kinase activity, respectively, have been observed in many types of human cancer. To explore the implication of PTEN and PIK3CA mutations in gastric tumorigenesis, we characterized the expression and mutation status of the genes in 126 gastric tissues and 15 cell lines. Expression of PTEN transcript was abnormally low in 5 of 15 (33%) cell lines and 20 of 55 (36%) primary carcinomas, whereas 0 of 71 noncancerous tissues including 16 benign tumors showed altered expression. Allelotyping analysis using an intragenic polymorphism (IVS4+109) revealed that 14 of 30 (47%) informative cases carried LOH of the gene, which is closely linked to low expression. The LOH rate was significantly higher in advanced tumors [12 of 19 (63%)] compared to early-stage tumors [2 of 11 (18%)] and more frequent in poorly differentiated tumors [9 of 13 (69%)] than well- or moderately differentiated tumors [5 of 17 (29%)]. Interestingly, however, none of the LOH tumors carried mutational disruption of the remaining allele, suggesting haploinsufficiency of PTEN in gastric tumorigenesis. Methylation studies revealed that PTEN pseudogene, but not PTEN, is methylated in cell lines and primary tumors, indicating that PTEN is not a target of epigenetic silencing in gastric cancers and that the pseudogene should be considered more carefully in methylation analysis of the PTEN promoter. Genomic amplification of PIK3CA was found in 9 of 15 (60%) cell lines and 20 of 55 (36.4%) primary tumors but in no noncancerous tissues. Furthermore, PIK3CA amplification was predominantly detected in tumors with no PTEN alterations, suggesting that mutations of PTEN and PIK3CA are mutually exclusive events in gastric tumorigenesis. Amplification of PIK3CA was strongly associated with increased expression of PIK3CA transcript and elevated levels of phospho-AKT. Collectively, our data reveal that 13 of 15 (87%) gastric cell lines and 31 of 55 (56%) primary carcinomas harbored either amplification of PIK3CA or abnormal reduction of PTEN. Mutually exclusive alterations of PTEN and PIK3CA also suggest that mutations of either gene could activate the PI3-kinase/AKT signaling pathway, which is directly linked to the malignant progression of gastric tumor cells. PMID- 12569556 TI - Mucosal immunoglobulin-A and -G responses to oncogenic human papilloma virus capsids. AB - Oncogenic human papilloma virus (HPV) infection is the most important risk factor for developing cervical cancer. It is known that serum antibody responses against these viruses are associated with persistent infection. We conducted an epidemiological study of 627 women to detect cervical mucosal immunoglobulin (Ig)A and IgG responses to oncogenic HPV capsids. Antibody reactivity and cervical HPV infection genotypes were examined by enzyme-linked immunosorbent assay (ELISA) using HPV types 16, 18, 31, and 45 virus-like particles, and a polymerase chain reaction-based method, respectively. HPV infection was defined as being positive for HPV DNA. Multivariate analysis revealed that a mucosal IgA response was associated with the HPV infection, whereas the IgG response was associated with high-grade cervical squamous intraepithelial lesions (SIL)/squamous cell cancer (SCC) and subject age (40-49 years). IgA was positive in 72% of women with oncogenic HPV infections, whereas IgG was positive in 64% of women with high-grade SIL/SCC. The longitudinal study demonstrated that the IgA response was elicited earlier than the IgG response, and the IgG response was barely induced in the preclinical HPV infection. However, once an IgG response was induced, it persisted longer after HPV clearance. The mucosal IgA response reflects current HPV infection, whereas an IgG response may be induced with the development of cervical lesions. PMID- 12569557 TI - Human papillomavirus infection as a prognostic factor in carcinomas of the oral cavity and oropharynx. AB - Although studies have established human papillomaviruses (HPVs) as a risk factor for oral and oropharyngeal cancer, it is not clear whether viral infection affects survival in head and neck malignancies. This investigation examined the relationship between HPV and survival in carcinomas of the oral cavity and oropharynx. Formalin-fixed, paraffin-embedded tumor specimens from 139 newly diagnosed cases were tested for HPVs by PCR and DNA sequencing. Patient and tumor characteristics were obtained from questionnaires, pathology reports and cancer registries. Odds ratios (ORs) and relative risks (RRs) were based on logistic and Cox regression models, respectively. HPVs were detected in 21% of the tumors; 83% were HPV-16. Greater risk of HPV infection was associated with males (OR = 2.9, 95% CI = 1.0-8.6), a history of oral-genital sex (OR = 4.2, 95% CI = 1.5-11.7), and oropharyngeal tumors (OR = 10.4, 95% CI = 3.5-31.2). As tobacco usage increased, the odds of HPV detection decreased (OR = 0.97/pack-year, 95% CI = 0.96-0.99). HPV infected patients had better overall survival (RR = 0.3, 95% CI = 0.1-0.8) than those with HPV-negative tumors. There was an interaction between gender and HPV for overall (p = 0.05) and disease-specific (p = 0.03) survival that suggested that HPV infected males had better prognosis than HPV-negative males, but this was not the case among females. HPV status was identified as an independent prognostic factor in oral and oropharyngeal cancers. This result appeared to be gender-specific, suggesting the need for further study of the interaction between HPV and gender on survival. PMID- 12569558 TI - Identification of a naturally processed HLA-A*0201 HPV18 E7 T cell epitope by tumor cell mediated in vitro vaccination. AB - Immunotherapy of HPV-associated disease such as cervical cancer is moving from preclinical investigation to clinical trials. The viral oncoproteins E6 and E7 are ideal target antigens because their expression is mandatory in HPV transformed tumor cells. T cells are the most important effector cells for therapeutic vaccination strategies. Therefore, the identification and characterization of HPV E6 and E7 T cell epitopes is necessary. Methods to date rely on screening for immunogenicity of peptides predicted by algorithms. Presentation of the identified peptides on tumor cells, however, needs to be confirmed. In our study, we have improved the method to identify peptide epitopes of HPV18 E7 that are actually presented by tumor cells. We induced allogeneic T cell lines by stimulation with HPV18-positive, CD80 and HLA-A*0201 transfected cervical cancer cells. Sensitized T cells were probed against an array of a HPV18 E7 20mer peptide-library. We found specific reactivity to one of the 20mer peptides. This sequence was then screened via algorithms for putative epitopes. One putative HLA-A2 restricted epitope was confirmed to bind to HLA-A2, to be immunogenic and to induce IFN gamma-release in ELISpot assays. Epitope-specific T cells were cytolytic toward autologous peptide pulsed targets and HPV18 transformed tumor cells. The identification of epitope-specific T cells in tumor infiltrating lymphocytes of a HPV18-positive HLA-matched cervical cancer patient suggests an in vivo relevance of the identified epitope. We suggest that our approach is advantageous over conventional methods, because it yields candidate peptides that are relevant CTL epitopes that are expressed, processed and presented by tumor cells. PMID- 12569559 TI - Expression and role of MICA and MICB in human hepatocellular carcinomas and their regulation by retinoic acid. AB - Natural killer (NK) cells are important effector cells for the first line of defense against tumor, but the mechanisms by which they recognize and kill human hepatocellular carcinoma (HCC) remains to be elucidated. Distant MHC class I homologs MICA and MICB are recently identified human ligands for NK cell activating receptor NKG2D. In our present study, MICA or MICB transcript was detected in 6 of 10 human hepatocellular carcinoma tissues, but not in the surrounding non-cancerous tissues. Immunohistochemical analysis showed that MICA/B were expressed in the tumor cells of the cancerous tissues. Huh7 and HepG2 hepatoma cells, but not Hep3B cells, substantially expressed MICA/B on their cell surface. MICA/B expressed on hepatoma cells contributed to their NK sensitivity, because Huh7 and HepG2 were less susceptible to NK cytolysis when MAb against MICA/B was added during the cytolysis assay. Of interest is the finding that retinoic acid upregulated expression of MICA/B in Huh7 and HepG2 cells. Retinoic acid-treated hepatoma cells induced IFN gamma production from cocultured NK cells and rendered themselves more susceptible to NK cells. This was clearly dependent on upregulation of MICA/B, because both the enhanced IFN gamma production and NK cytolysis were completely abolished by MAb-mediated masking of MICA/B. These results suggest that MICA/B, expressed on a subset of human HCCs, may play an important role in their susceptibility to NK cells. Furthermore, retinoic acid can function as a modulator of MICA/B expression and thereby further activate NK cells. PMID- 12569560 TI - A CD4+, HLA-DR7-restricted T-helper lymphocyte clone recognizes an antigen shared by human malignant melanoma and glioma. AB - CD4(+) Th cells that are restricted by MHC class II molecules play an important role in the induction of antitumor immune responses. We have established a stable CD4(+) Th cell clone (Th35-1A) from the PBMCs of a patient with primary cutaneous melanoma. The Th cell clone is noncytolytic and proliferates specifically in the presence of irradiated autologous melanoma cells or autologous EBV-transformed B cells pulsed with melanoma tumor cell lysates. Th35-1A produces IFN-gamma (a Th1 type cytokine) after autologous tumor cell stimulation, and its proliferative reactivity is HLA class II-restricted. Th cells showed helper activity for PWM responses of PBMCs. Using a panel of HLA class II-matched and unmatched EBV-B cells as APCs and allogeneic melanoma tumor cell lysate as stimulant, DR7 was delineated as the HLA class II restriction element used by the Th cell clone. In agreement with these results, transfection of an allogeneic melanoma cell line with HLA-DR7 isolated from autologous EBV-B cells rendered the cell line stimulatory for Th35-1A cells. Specificity studies using autologous EBV-B cells (EBV-B35) pulsed with a panel of allogeneic tumor cell lysates of various tissue origins indicated that the Th cell clone recognizes an antigen shared by melanoma and glioma cells. The availability of the Th cell clone may lead to the development of new therapies against melanoma, using adoptive Th cell transfer and/or active immunization with a shared Th cell antigen. PMID- 12569561 TI - Abnormal expression pattern of cyclin E in tumour cells. AB - The expression pattern of cyclin E during the cell cycle was studied in normal and tumour cells in culture and in tumour biopsies. This pattern was found to be abnormal in tumour cells. A triple immunostaining protocol, digital microscopy and image analysis were used to find the position of the individual cells in the cell cycle and to measure the nuclear cyclin E levels. In normal cells, the number of cyclin E-positive cells decreased rapidly when the cells entered the S phase. In the tumour cell lines, cyclin E was not downregulated in early S-phase, as in normal cells. Instead the number of cyclin E-positive cells remained high throughout S-phase, and the cyclin E staining intensity per cell often increased during S-phase. In about half of the analysed tumour cell lines, many cells stained positive for cyclin E even in the G(2)-phase. This abnormal expression over the cell cycle of cyclin E was also found in tumour biopsies from cervical, breast and prostatic carcinomas, even though it varied greatly between individual tumours. In some tumours, the expression pattern of cyclin E was similar to that of normal cells in culture, whereas in others high cyclin E levels could be seen in S-phase cells, as in the transformed cell lines. A high percentage of cells expressing cyclin E during S- or G(2)-phase was found to be related to poor outcome (p < 0.025) in a small group of cervical carcinoma patients (n = 12). PMID- 12569562 TI - Anti-tumor effect and increased survival after treatment with [177Lu DOTA0,Tyr3]octreotate in a rat liver micrometastases model. AB - Peptide receptor scintigraphy with [(111)In-DTPA(0)]octreotide (a stabilized radiolabeled somatostatin (SS) analogue, OctreoScan) is widely used for the visualization and staging of somatostatin receptor-positive tumors. The application of likewise somatostatin analogues as vehicle for the deliverance of radionuclides to somatostatin receptor-positive targets are now in use for peptide receptor-targeted radionuclide therapy (PRRT). Currently preclinical and clinical investigation are ongoing trying to find the optimal combination of radionuclide and ligand. The anti-tumoral effects of such combinations, like [90Y DOTA degrees, Tyr(3)]octreotide and [(177)Lu-DOTA degrees, Tyr(3)]octreotate, on SSR-positive solid tumors have been reported. In this study we present the anti tumor effects of (177)Lu-DOTA-tate on: a) a single SSR-positive cell model and b) on a SSR-positive tumor in a rat liver micrometastatic model, mimicking disseminated disease. (177)Lu-DOTA-tate showed anti-tumoral effects in both cases and significant survival in the PRRT-treated rats. (177)Lu-DOTA-tate is a very promising new treatment modality for SSR-positive tumors, including disseminated disease. PMID- 12569564 TI - BACH1 517C-->T transition impairs protein translocation to nucleus: a role in breast cancer susceptibility? PMID- 12569563 TI - Polyamine biosynthesis inhibition enhances HSV-1 thymidine kinase/ganciclovir mediated cytotoxicity in tumor cells. AB - Cancer gene therapy with the aid of herpes simplex virus type 1 thymidine kinase gene (HSV-TK) and anti-herpes drug ganciclovir (GCV) has been widely used and its efficacy has been demonstrated in a variety of different malignant cells and animal tumor models. It is also commonly accepted, however, that this gene therapy regimen needs to be enhanced for a true clinical success. We studied whether polyamine biosynthesis inhibition by 2-difluoromethylornithine (DFMO), a clinically tested and well-tolerated chemotherapeutic drug, can increase the cytotoxicity of HSV-TK/GCV in 9L rat glioma cells. Our initial experiments showed that polyamine depletion actually protected the cells from cytotoxicity if GCV treatment was started too early after removal of DFMO. Analyses of cell growth, intracellular polyamine pools and cell cycle phase distribution suggested that later initiation of GCV treatment would be more beneficial due to increased proportion of cells in the middle of the cell cycle S phase. When the cells were exposed to GCV 3 or 4 days after removal of DFMO from growth medium, the cytotoxicity was increased up to 2.5-fold. We also verified whether cell cycle blockage per se could yield similar effect as DFMO. Our results from serum deprivation experiments showed that, despite of apparent cell growth and cell cycle phase distribution effects, serum starvation was weaker enhancer of HSV TK/GCV cytotoxicity than DFMO. Finally, the general utility of HSV-TK/GCV + DFMO combination was tested in another tumor cell type, human prostate carcinoma cell line DU-145. DFMO sensitized these cells to HSV-TK/GCV cytotoxicity, but the effect was less prominent than in 9L cells. In conclusion, we have demonstrated that a correctly timed induction of DFMO-mediated polyamine biosynthesis inhibition can enhance the efficiency of HSV-TK/GCV gene therapy in vitro. The observed synergistic effect is potentially useful in clinical trials because, as opposed to use of other cell cycle-altering drugs, DFMO has already been tested in the treatment of human tumors and used as chemo preventive regimen with excellent tolerability. PMID- 12569566 TI - Differentially expressed transcripts in neoplastic hepatic nodules and neonatal rat liver studied by cDNA microarray analysis. AB - The molecular mechanisms underlying hepatocarcinogenesis remain unclear. Wistar rats treated with 2-AAF develop hepatocarcinoma in histologically well characterised stages. In our study, cDNA microarrays were used to measure the expression of 3,000 genes during the progression of liver carcinogenesis in persistent neoplastic nodules. Because tumours frequently revert into a more poorly differentiated phenotype, we also studied the expression of the same set of transcripts in neonatal rat liver. Approximately 2,000 transcripts gave a detectable signal in experiments comparing gene expression in nodules and control tissue. Approximately 8% of these were identified as differentially expressed in liver nodules. The differentially expressed genes fell into several categories with putative or demonstrated roles in signal transduction, metabolism, detoxification, cell-structure and transport. Many of the differentially expressed genes in nodules were not previously known to be regulated during liver carcinogenesis. A universal transcript profile for gene expression in hepatic liver nodules and neonatal liver has been created. PMID- 12569567 TI - Breast cancer cell line proliferation blocked by the Src-related Rak tyrosine kinase. AB - Rak is a 54 kDa protein tyrosine kinase originally isolated from breast cancer cells and expressed in epithelial cells. It resembles the protooncogene Src structurally but lacks an amino-terminal myristylation site and localizes to the nuclear and perinuclear regions of the cell. We report here that expression of Rak in 2 different breast cancer cell lines inhibits growth and causes G(1) arrest of the cell cycle. This growth inhibition is kinase-dependent but does not require the Rak SH2 or SH3 domain. Rak also binds to the pRb tumor-suppressor protein but inhibits growth even in cells that lack pRb. These results suggest that Rak regulates cell growth by phosphorylating perinuclear proteins and has a function that is distinct from the Src-related kinase family. PMID- 12569568 TI - Overexpression of cadherins suppresses pulmonary metastasis of osteosarcoma in vivo. AB - Osteosarcoma by nature shows aggressive pulmonary metastasis; however, the underlying molecular mechanisms remain unclear. We previously showed that N cadherin and cadherin-11 (OB-cadherin), which are highly expressed in normal osteoblasts, are anomalously expressed in human osteosarcoma (Kashima et al., Am J Pathol 1999;155:1549-55). In the present study, we examined the role of cadherins in osteosarcoma metastasis using the mouse osteosarcoma cell line Dunn and its highly metastatic subline LM8. Oligonucleotide array and RT-PCR analyses demonstrated that Dunn and LM8 cells did not express appreciable levels of several members of the cadherin family, and Western blot analysis confirmed that Dunn and LM8 cells did not express P-cadherin, E-cadherin, N-cadherin or cadherin 11 protein. We therefore investigated the functional consequences of cadherin overexpression on cell migration and in vivo metastatic potential of LM8 cells. Several LM8 clones were isolated which expressed exogenous N-cadherin and cadherin-11 localized to the cell membrane and able to bind to beta-catenin. Overexpression of N-cadherin or cadherin-11 in LM8 cells did not affect cell proliferation but caused an inhibitory effect on cell migration in vitro. In vivo analysis showed that N-cadherin- and cadherin-11-overexpressing cells exhibited a marked reduction in their ability to form pulmonary metastases, with significant decreases in lung weight and the number and weight of metastatic lesions, as well as the size and weight of primary lesions at the s.c.-inoculated site. These observations demonstrate that disruption of N-cadherin- and cadherin-11-mediated cell-cell adhesion is critical in the pulmonary metastasis of osteosarcoma. PMID- 12569569 TI - Letrozole as a potent inhibitor of cell proliferation and expression of metalloproteinases (MMP-2 and MMP-9) by human epithelial breast cancer cells. AB - P450 aromatase catalyzes the conversion of androgens to estrogens and plays a key role in the cell growth of hormone-dependent breast cancer in postmenopausal women. On the other hand, matrix metalloproteinases (MMPs), which can degrade almost all components of the extracellular matrix, play a crucial role in tumor cell invasion and cancer metastasis. In the present study the effect of letrozole on cell proliferation of estrogen receptor (ER)-positive MCF-7 human epithelial breast cancer and MCF-12A human mammary epithelial cells was studied. The effect of letrozole on the in vitro release of MMPs, particularly type IV collagenases (MMP-2 and MMP-9), by the ER-positive MCF-7 cells was also investigated, using a solid-phase method of high sensitivity and accuracy. Using RNA isolates from cell lines MCF-7 and MCF-12A, reverse transcriptase-polymerase chain reaction analysis revealed that only MCF-7 cells express the P450 aromatase gene. Study of the effects of letrozole alone and the hormones 17-beta-estradiol, testosterone and 4 androstene-3, 17-dione in the presence and absence of letrozole on cell growth at the DNA synthesis level showed that letrozole significantly suppressed the endogenous aromatase-induced proliferation of MCF-7 cells. The majority of MMPs secreted by MCF-7 cells were identified in their pro-forms, which was in accordance with the low metastatic potential determined for these cells. After treatment of cells with letrozole (10 nM) for 24 and 48 h, significant inhibition of MMP levels was obtained. Furthermore, concurrent treatment of MCF-7 cells with 17-beta-estradiol in the presence of letrozole significantly suppressed the estradiol-induced stimulation of MMP levels. The data obtained suggest that letrozole is a potent in vitro inhibitor of cell proliferation and of type IV collagenases expressed by ER-positive MCF-7 cells and may be of value for suppressing breast tumor growth and invasiveness. PMID- 12569570 TI - Reduced expression of transforming growth factor-beta receptors is an unfavorable prognostic factor in human esophageal squamous cell carcinoma. AB - Transforming growth factor-beta (TGF-beta) inhibits epithelial cell proliferation. Inactivation of the TGF-beta signaling pathway is thought to play a role in tumorigenesis. Our purpose was to clarify the correlation between TGF beta receptors or TGF-beta 1 expression and the clinicopathologic characteristics of patients with esophageal squamous cell carcinoma (SCC). Immunohistochemical staining for TGF-beta type I receptor (TGF-beta R-I), TGF-beta R-II and TGF-beta 1 was performed on surgical specimens obtained from 80 patients with esophageal SCC. Preoperative plasma TGF-beta1 levels were measured and correlated with pathologic features and clinical outcomes. Expression of TGF-beta R-I and TGF beta R-II was reduced in 43 (53.8%) and 23 (28.8%) specimens, respectively. TGF beta 1 was overexpressed in 29 (36.3%). Reduced expression of TGF-beta R-I and TGF-beta R-II showed a significant association with depth of invasion (p = 0.0015 and p = 0.0012), lymph node metastasis (p = 0.0309 and p = 0.0059) and pathologic stage (p = 0.0103 and p = 0.0401). Overexpression of TGF-beta 1 had a significant association with depth of invasion only (p = 0.0335). Reduced expression of TGF beta R-I and TGF-beta R-II was correlated with cancer-specific survival (p = 0.0324 and p = 0.0243). The mean preoperative plasma TGF-beta 1 level was 10.5 +/ 0.8 ng/ml in patients with esophageal carcinoma and was significantly higher compared to healthy controls (p < 0.01). We demonstrate that reduced expression of TGF-beta receptors in esophageal SCC appears to be correlated with depth of invasion, lymph node metastasis, pathologic stage and poor prognosis. TGF-beta receptor expression may play a key role in the progression of this cancer. PMID- 12569571 TI - Inhibition of heparanase activity and heparanase-induced angiogenesis by suramin analogues. AB - Heparanase, a heparan sulfate-specific endo-beta-D-glucuronidase, plays an important role in tumor cell metastasis through the degradation of extracellular matrix heparan sulfate proteoglycans (ECM HSPG). Heparanase activity correlates with the metastatic propensity of tumor cells. Suramin, a polysulfonated naphthylurea, is an inhibitor of heparanase with suramin analogues shown to possess antiangiogenic and antiproliferative properties. We investigated the effects of selected suramin analogues (NF 127, NF 145 and NF 171) on heparanase activity and heparanase-driven angiogenesis. Studies of the ability of cellular extracts and purified heparanase from human, highly invasive and brain-metastatic melanoma (70W) cells revealed that heparanase expressed by these cells was effectively inhibited by suramin analogues in a dose-dependent manner. These analogues possessed more potent heparanase inhibitory activities than suramin: The concentrations required for 50% heparanase inhibition (IC(50)) were 20-30 microM, or at least 2 times lower than that for suramin. One hundred percent inhibition was observed at concentrations of 100 microM and higher. Of relevance, these compounds significantly decreased (i) the invasive capacity of human 70W cells by chemoinvasion assays performed with filters coated with purified HSPG or Matrigel trade mark, and (ii) blood vessel formation by in vivo angiogenic assays, thus linking their antiangiogenic properties with impedance of heparanase induced angiogenesis. Specifically, inhibition of invasion by NF 127, NF 145 and NF 171 was found at 10 microM concentrations of compounds with a significant decrease of invasive values at concentrations as low as 1.5 microM. In addition, NF 127, NF 145 and NF 171 promoted nearly complete inhibition of heparanase induced angiogenesis at values ranging from 236 microM (for NF 145) to 362 microM (for NF 127). These results further emphasize the importance of heparanase in invasive and angiogenic mechanisms and the potential clinical application of heparanase inhibitors such as suramin analogues in cancers and angiogenesis dependent diseases. PMID- 12569572 TI - PTEN/MMAC1 gene mutation is a rare event in soft tissue sarcomas without specific balanced translocations. AB - The tumor suppressor gene PTEN/MMAC1 was identified on chromosome 10q23.3, which is homozygously deleted in many human malignancies. The loss of chromosome 10q was also frequently reported in some types of soft tissue sarcomas. Our study was designed to investigate the frequency of PTEN/MMAC1 gene mutation and to evaluate the role of the PTEN/MMAC1 gene in the tumorigenesis of soft tissue sarcomas without specific balanced translocations. We analyzed 51 cases of soft tissue sarcomas without specific balanced translocations for PTEN/MMAC1 mutations by polymerase chain reaction-single strand conformation polymorphism and direct sequencing. Mutations in the PTEN/MMAC1 gene were found in only 2 cases (3.9%). Both tumors with PTEN/MMAC1 mutation were leiomyosarcomas arising from the retroperitoneum and inferior vena cava, respectively. Two of 3 leiomyosarcomas arising from the intra-abdominal cavity examined harbored mutations of this tumor suppressor gene. This result suggests that leiomyosarcomas derived from the intra abdominal cavity might have different tumorigenesis from those of an extremity or the trunk, from the viewpoint of PTEN/MMAC1 mutation, although PTEN/MMAC1 gene mutations are rare event in these soft tissue sarcomas. PMID- 12569573 TI - Regulation of the H19 imprinting gene expression in human nasopharyngeal carcinoma by methylation. AB - In East Asia and Singapore, the human nasopharyngeal carcinoma (NPC) presented clinically is mainly of the undifferentiated type. In contrast, the well differentiated squamous NPC is more commonly detected in the West. To study the potential differences in carcinogenesis between undifferentiated and differentiated human NPC, we employed cDNA microarrays to isolate genes that might be specific for human undifferentiated NPC. One of the genes identified to be specifically upregulated in the undifferentiated human NPC cell line CNE-2 is the human imprinting gene H19. Interestingly, H19 is not expressed in the well differentiated human HK1 NPC cells. Northern blot and in situ hybridization analyses also confirmed that the H19 gene is strongly expressed in the undifferentiated CNE-2 human NPC cell line but not in the well-differentiated HK1 human NPC cell line. In situ hybridization and reverse transcriptase-polymerase chain reaction also demonstrated that H19 is specifically expressed in NPC biopsies and not in non-NPC human tissue biopsies. Furthermore, we demonstrated that deregulation of H19 gene expression in the well-differentiated human HK1 NPC cells could be induced by the hypomethylation of CpG sites of the H19 promoter region. Hypermethylation of gene promoter regions might therefore be an important epigenetic event that plays a role in the differentiation of human NPC cells and the transcriptional silencing of imprinted genes. PMID- 12569574 TI - Effects of granulocyte-macrophage colony-stimulating factor and foreign helper protein as immunologic adjuvants on the T-cell response to vaccination with tyrosinase peptides. AB - Immunologic adjuvants are used to augment the immunogenicity of MHC class I restricted peptide vaccines, but this effect has rarely been systematically evaluated in a clinical trial. We have investigated, in a phase I study, whether addition of the 2 adjuvants GM-CSF and KLH can enhance the T-cell response to MHC class I peptide vaccines. Forty-three high-risk melanoma patients who were clinically free of disease received 6 vaccinations with MHC class I-restricted tyrosinase peptides alone, with either GM-CSF or KLH or with a combination of both adjuvants. The primary end point was induction of tyrosinase-specific T cells, and serial T-cell monitoring was performed in unstimulated peripheral blood samples before and after the second, fourth and sixth vaccinations by ELISPOT assay. Tyrosinase-specific IFN-gamma-producing T cells were detected as early as 2 weeks after the second vaccination in 5 of 9 patients vaccinated with tyrosinase peptides in combination with GM-CSF and KLH but not in any patient vaccinated with tyrosinase peptides without adjuvants or in combination with either adjuvant alone. After 6 vaccinations, tyrosinase-specific T cells were found in patients immunized with peptides either without adjuvants (3 of 9 patients) or in combination with the single adjuvant GM-CSF (4 of 9 patients) but not with KLH (0 of 10 patients). Our results suggest that addition of either GM CSF or KLH as a single adjuvant has little impact on the immunogenicity of tyrosinase peptides. The combined application of GM-CSF and KLH was associated with early induction of T-cell responses. PMID- 12569575 TI - Dysregulated PTEN-PKB and negative receptor status in human breast cancer. AB - Recent studies demonstrate that abnormalities in PTEN may be one of the most frequent genetic events observed in human cancers. PTEN dysfunction leads to tumorigenesis through unopposed survival signals mediated via activated protein kinase B (PKB), which may also be associated with hormone-independence. We therefore investigated the relationship between PTEN-PKB and receptor status in human breast cancer. Several molecular variables, including immunohistochemical staining for PTEN, PKB (phosphorylated on ser473), p53 and p21 were evaluated. The p53 gene was sequenced from exons 2-11. Seventy-eight participants in a randomised breast cancer trial served as the cohort for our study. Twenty-eight of 77 (36%) patients' tumours demonstrated absent or reduced PTEN expression; 17 of 78 (22%) tumours over-expressed P-PKB. A significant inverse relationship was observed between reduced PTEN and increased P-PKB expression. Reduced PTEN also correlated with reduced ER or PR expression. None of the molecular variables correlated with survival. ER and PR negative tumours, however, experienced a significantly inferior disease-free survival than other ER/PR status tumours. Immunohistochemical analyses of ER expression in mammary carcinomas arising in PTEN heterozygous knockout mice did not demonstrate a reduction in ER immunoreactivity, in comparison to wild-type mice. Our data demonstrate that the PTEN-PKB pathway is abnormal in approximately 1/3 of lymph node negative breast cancer. Dysregulated PTEN-PKB was also associated with reduced ER/PR expression, but this does not appear to be a simple direct causal relationship. These observations support the contention that dysregulation in PTEN-PKB contributes to disease progression and hormone resistance of human breast cancer. PMID- 12569576 TI - Differential expression of genes induced by resveratrol in LNCaP cells: P53 mediated molecular targets. AB - Prostate cancer prevention by key elements present in human nutrients derived from plants and fruits has been confirmed in various cell cultures and tumor models. Resveratrol (RE), a phytoalexin, induces remarkable inhibitory effects in prostate carcinogenesis via diverse cellular mechanisms associated with tumor initiation, promotion and progression. Earlier studies have shown that RE alters the expression of genes involved in cell cycle regulation and apoptosis, including cyclins, cdks, p53 and cdk inhibitors. However, most of the p53 controlled effects related to the role of RE in transcription either by activation or repression of a sizable number of primary and secondary target genes have not been investigated. Our study examined whether RE activates a cascade of p53-directed genes that are involved in apoptosis mechanism(s) or whether it modifies the androgen receptor and its co-activators directly or indirectly and induces cell growth inhibition. We demonstrate by DNA microarray, RT-PCR, Western blot and immunofluorescence analyses that treatment of androgen sensitive prostate cancer cells (LNCaP) with 10(-5) M RE for 48 hr downregulates prostate-specific antigen (PSA), AR co-activator ARA 24 and NF-kB p65. Altered expression of these genes is associated with an activation of p53-responsive genes such as p53, PIG 7, p21(Waf1-Cip1), p300/CBP and Apaf-1. The effect of RE on p300/CBP plays a central role in its cancer preventive mechanisms in LNCaP cells. Our results implicate activation of more than one set of functionally related molecular targets. At this point we have identified some of the key molecular targets associated with AR and p53 target genes. These findings point to the need for further extensive studies on AR co-activators, such as p300, its central role in post-translational modifications such as acetylation of p53 and/or AR by RE in a time- and dose-dependent manner at different stages of prostate cancer that will fully elucidate the role of RE as a chemopreventive agent for prostate cancer in humans. PMID- 12569577 TI - Human papillomavirus infection among women in South and North Vietnam. AB - The incidence rate of invasive cervical carcinoma (ICC) is 4-fold higher in Ho Chi Minh City, in the South of Vietnam, than in Hanoi, in the North. Thus, we explored the prevalence of and the risk factors for human papillomavirus (HPV) infection in these 2 areas. A population-based random sample of married women aged 15-69 years were interviewed and had a gynaecological examination in the urban district of Ho Chi Minh City and in a peri-urban district in Hanoi. HPV DNA detection was performed using a GP5+/6+ primer-mediated PCR enzyme immunoassay. A total of 922 women from Ho Chi Minh and 994 from Hanoi, for whom a Pap smear and HPV-status were available, were evaluated. HPV DNA was detected among 10.9% of women in Ho Chi Minh City and 2.0% in Hanoi (age standardized prevalence, world standard population: 10.6% and 2.3%, respectively). In the 2 areas combined, 30 different HPV types were found, the most common being HPV 16 (in 14 single and 18 multiple infections), followed by HPV 58, 18 and 56. A peak of HPV DNA detection in women younger than age 25 was found in Ho Chi Minh City (22.3%) but not in Hanoi. Major risk factors for HPV DNA detection were indicators of sexual habits, most notably the presence of HSV-2 antibodies, nulliparity and the current use of oral contraceptives. Women in Hanoi showed the lowest HPV prevalence ever reported so far, suggesting that HPV has not spread widely in this population. As expected, HPV prevalence in a population seemed to be closely correlated with ICC incidence rates. PMID- 12569578 TI - Meat, fish and egg intake and risk of breast cancer. AB - Intakes of animal protein, meat, and eggs have been associated with breast cancer incidence and mortality in ecological studies, but data from long-term prospective studies are limited. We therefore examined these relationships in the Nurses' Health Study. We followed 88,647 women for 18 years, with 5 assessments of diet by food frequency questionnaire, cumulatively averaged and updated over time. We calculated the relative risks (RR) and 95% confidence intervals (95% CI) for risk of developing invasive breast cancer, over categories of nutrient and food intake. During follow-up, 4,107 women developed invasive breast cancer. Compared to the lowest quintile of intake, the RR and 95% CI for the highest quintile of intake were 1.02 (0.92-1.14) for animal protein, 0.93 (0.83-1.05) for red meat and 0.89 (0.79-1.00) for all meat. Results did not differ by menopausal status or family history of breast cancer. We found no evidence that intake of meat or fish during mid-life and later was associated with risk of breast cancer. PMID- 12569579 TI - Ovulation and risk of epithelial ovarian cancer. AB - Incessant ovulation is thought to be one of the primary causes of epithelial ovarian cancer. However, the effects of ovulation at different ages and of the various exposures or events that suppress ovulation have not been established. We used data from an Australian case-control study of 791 ovarian cancer cases and 853 controls to examine the effect of ovulation on ovarian cancer risk. The total number of lifetime ovulations was calculated using information provided in a monthly contraceptive/reproductive calendar, as well as incorporating other information such as average menstrual cycle length. An increase of 1 year's worth of ovulation was associated with a 6% increase in risk of ovarian cancer (95% confidence interval [CI] = 4-8%). Ovulations in the 20-29-year age group were associated with the greatest risk, with a 20% increase in risk associated with each year of ovulation during this age period (95% CI = 13-26%). When the effects of different exposures that suppress ovulation were compared, there was an indication that some factors may have a greater effect than others. These findings support the theory that incessant ovulation is a major contributor to the occurrence of ovarian cancer and suggest that ovulations during the 20s may be those most associated with disease risk. PMID- 12569580 TI - Rapamycin inhibits proliferation of human neuroblastoma cells without suppression of MycN. AB - MYCN and insulin-like growth factor (IGF) system are important for the pathogenesis and development of neuroblastoma. We previously reported evidence of a direct linkage between MycN and the IGF system in KP-N-RT human neuroblastoma cells, where IGF-I induced both MycN expression at the RNA level and G1-S cell cycle progression through the IGF-I receptor (IGF-IR)/ MEK/ mitogen-activated protein kinase (MAPK) pathway (A. Misawa et al., Cancer Res, 2000; 60:64-9). Our data also showed the possibility of a potent IGF-IR downstream signal cascade that accelerates progression into the S-phase, other than the MAPK pathway. In this study, we further investigated the role of this alternative pathway in the growth of neuroblastoma cells. A phosphoinositide 3-kinase (PI3K) inhibitor wortmannin blocked IGF-I-mediated induction of MycN. Our data suggest that the inhibition of MycN by wortmannin was transmitted through the MAPK pathway. Progression of the cell cycle from G1 to S phase was inhibited up to 90% by wortmannin or rapamycin, an inhibitor of mTOR, which acts downstream of PI3K. Despite its effects on induction of MycN and on progression through S phase, wortmannin did not block proliferation of neuroblastoma cells. On the other hand, rapamycin inhibited both IGF-I-induced cell cycle progression and cell proliferation in complete medium, although it had no effect on IGF-I-mediated MycN induction. Our study indicates maintenance of cell proliferation requires mTOR function, which is independent of MycN induction in human neuroblastoma cells. PMID- 12569581 TI - Antiproliferative effects of the histone deacetylase inhibitor FR901228 on small cell lung cancer lines and drug-resistant sublines. AB - FR901228 is a novel histone deacetylase (HDAC) inhibitor, and its antiproliferative effects on non-small cell lung cancer cells have been shown in vitro. However, there have been no reports concerning the effects on small-cell lung cancer (SCLC). We have recently demonstrated that the HDAC inhibitors trichostatin A and sodium butyrate inhibit expression of the catalytic subunit of telomerase (hTERT) mRNA and telomerase activity in prostate cancer cells. The present study was designed to evaluate the effects of FR901228 on proliferation and telomerase activity in SCLC cells in vitro. FR901228 at 5 to 10 nM increased the fraction of cells in the G(2)/M and sub-G(1) phases of the cell-cycle, and inhibited the growth of H69, H526 and H82 cell lines. The expression of hTERT mRNA was inhibited 6 hr after treatment, prior to obvious inhibition of cell growth or cell-cycle distribution shifts. The inhibition of hTERT mRNA expression and telomerase activity was not a consequence of cell-growth arrest or apoptosis. Cycloheximide blocked the suppression of hTERT mRNA induced by FR901228, and the inhibition of hTERT mRNA by FR901228 required newly synthesized proteins. FR901228 also effectively inhibited growth of etoposide-resistant UMCC-1/VP-16, irinotecan-resistant PC-6/SN2-5H and cisplatin-resistant H526/CDDP cells having decreased expression of hTERT mRNA and telomerase activity, as well as their parental cells. This implies that SCLC resistant to these key drugs are not cross resistant to FR901228. The present study suggests that FR901228 may be a promising drug for chemotherapy of cancers including SCLC, even for refractory or relapsing tumors after conventional chemotherapy. PMID- 12569582 TI - Alpha-tocopherol protects against cisplatin-induced toxicity without interfering with antitumor efficacy. AB - Our aim was 2-fold: to investigate the role of alpha-tocopherol supplementation on the antitumor activity of DDP and to evaluate the effect of alpha-tocopherol on the survival and neurotoxicity of DDP-treated mice. Experiments performed on the M14 human melanoma line demonstrated that alpha-tocopherol supplementation did not influence the efficacy of DDP; the inhibition of cell survival and of the in vivo tumor growth after treatment with alpha-tocopherol and DDP combination was similar to that observed after DDP alone. Conversely, alpha-tocopherol was also able to increase survival of mice treated with a high dose of DDP. While DDP alone produced death in about 70% of mice, the combination reduced deaths to about 30%. Analysis of oxidative stress markers and peroxidative damage in organs indicated that the protective effect of alpha-tocopherol was mainly related to its antioxidant activity. A significant increase in the concentration of TBARS and decreased PUFAs and catalase activity were observed after DDP treatment, while with alpha-tocopherol the levels of these markers were comparable to those observed in untreated mice. Histologic analysis performed on peripheral nerve revealed that alpha-tocopherol also protected mice from severe neurologic damage induced by DDP treatment. In conclusion, our results demonstrate that alpha tocopherol protects against the systemic toxicity and neurotoxicity induced by DDP without interfering with its antitumor activity and suggest that this combination is a promising strategy to improve the therapeutic index of DDP-based chemotherapy. PMID- 12569583 TI - The effect of aging on the incidence of Kaposi's sarcoma among HIV-positive individuals with known dates of seroconversion. PMID- 12569584 TI - Clearance of cervical human papillomavirus infections. PMID- 12569585 TI - Association of first ocular melanoma with subsequent cutaneous melanoma: results from the Swedish Family-Cancer Database. PMID- 12569586 TI - Ocular melanoma and cutaneous melanoma. PMID- 12569588 TI - Compliance with breast-conservation standards for patients with early-stage breast carcinoma. AB - BACKGROUND: Multiple treatment guidelines and practice standards have been developed regarding the management of patients with breast carcinoma. Few evaluations of the penetration and utility of these practice standards have been performed. In 1992, the American College of Surgeons (ACOS), the American College of Radiology, the College of American Pathologists, and the Society of Surgical Oncology collaborated in establishing standards for breast-conservation treatment (BCT). The authors sought to determine whether practice patterns for patients with breast carcinoma who underwent BCT were consistent with these standards 2 years after their dissemination and to establish whether compliance varied by the same patient and hospital variables that predicted for the amount of BCT performed. METHODS: A study specific questionnaire was circulated to cancer registrars through the Commission on Cancer of the ACOS asking them to submit reports on patients with Stage I and II breast carcinoma who were diagnosed in 1994. Eight hundred forty-two predominantly community hospitals throughout the United States responded, yielding a total of 16,643 analyzable patients. The frequency of compliance to the 1992 published practice standards for 7097 patients who received BCT was determined. The variation in compliance rates by patient age, race, and insurance status and the treating hospital's geographic locations and cancer programs were evaluated. RESULTS: Of the 22 standards that were evaluated in the areas of preoperative mammography (2 standards), labeling of the surgical specimen (3 standards), pathology report content (10 standards), radiation after lumpectomy (6 standards), and systemic therapy for patients with positive lymph nodes (1 standard), treatment adherence was > or = 80% for 16 standards (73%). Poor compliance was demonstrated for six standards: the documentation of an abnormality's size in the mammogram report, labeling the lumpectomy specimen with the affected quadrant of the breast, spatial orientation of the lumpectomy specimen and inclusion of lymphatic/vascular invasion, ductal carcinoma in situ, and macroscopic margin assessment in the pathology report. Variation in compliance to a standard occurred frequently across the type of hospital cancer program and geographic region (77% for both), and variation occurred less across the patient variables of age (32%), race (41%), and payer (23%). There was not a pattern of more frequent compliance among variables associated with more BCT use. CONCLUSIONS: Large-scale evaluation of the penetration of treatment standards is feasible. For patients who underwent lumpectomy, practice appeared to be consistent (> or = 80% compliance), with 73% of 22 treatment standards evaluated. The standards with poor compliance represent areas for targeted physician education and reevaluation. Significant differences in adherence to a standard were seen frequently based on a hospital's geographic location and type of cancer program. This emphasizes the importance of adequate dissemination of treatment standards to ensure penetration into medical practices of all types. PMID- 12569589 TI - Breast boost: are we missing the target? AB - BACKGROUND: Randomized trials have shown improved local control with the use of a breast boost for patients given breast-conserving treatment for breast carcinoma. Although the use of a breast boost is routine practice, no standard technique has been established. The authors compared the commonly used clinical technique with a technique based on computed tomography (CT) imaging of surgical clips in the tumor bed. METHODS: Thirty patients underwent CT simulation for postoperative radiation treatment planning as part of breast conservation therapy. During simulation, a CT-compatible wire was placed on the patient's skin, outlining the intended electron boost field; an electron boost volume (EBV) was generated by contouring the tissue underlying the wire. Also contoured was a CT-based clinical target volume (CTV) using surgical clips and postsurgical changes in the tumor bed as a guide. A planning target volume (PTV) was generated using a 1 cm margin around the CTV. An electron beam treatment plan was generated for each technique using the FOCUS three-dimensional treatment planning system. Dose-volume histograms (DVH) were generated to determine the fraction of the PTV receiving 90% of the prescribed dose if treatment was delivered using the EBV. In addition, DVH analysis was done to determine the volume of normal tissue unnecessarily irradiated when using the EBV. RESULTS: Although the electron cone size remained unchanged in most patients for both EBV and PTV, the isocenter differed more than 1 cm in the medial-lateral direction in 5 patients and in the cephalocaudal direction in 12 patients. The en face gantry angle differed for most patients. On average, only 51% (range, 27-79%) of the PTV received 90% or more of the prescribed dose when the electron plan was generated using the EBV (P < 0.0001). Ten patients received the prescription dose to less than 50% of the PTV. Mean volume of normal tissue receiving more than 50% of prescribed dose was 64.5 cm(3) (range, 24-119 cm(3)). CONCLUSIONS: Clinical delineation of the tumor bed not only carries a significant risk of missing the target, but unnecessarily treats breast tissue that may otherwise be spared. Better delineation of the tumor bed, which optimizes coverage of the target volume and spares normal breast tissue, has the potential to improve both local control and cosmetic outcome. The authors recommend the use of surgical clips to delineate the target volume, followed by CT-based treatment planning, accounting for not only microscopic disease, but also organ motion and daily setup error. PMID- 12569590 TI - Does local recurrence affect the rate of distant metastases and survival in patients with early-stage breast carcinoma treated with breast-conserving therapy? AB - BACKGROUND: The purpose of the current analysis was to evaluate the impact of local recurrence (LR) on the development of distant metastases (DM), overall survival (OS), and cause specific survival (CSS) in patients with early-stage breast carcinoma who underwent conservative surgery (CS) and received postoperative radiotherapy (RT). METHODS: Between 1980 and 1995, 1169 patients underwent CS and received RT. All patients were followed for > 1 year and had < or = 4 lymph nodes involved with disease. The median duration of follow-up was 7.7 years. A Cox proportional hazards model was performed to evaluate the effect of LR on the development of DM and CSS. A matched-pair analysis that controlled for multiple prognostic factors also was performed comparing the outcomes of patients with and without LR. RESULTS: The LR rate was 11% at 12 years. For the entire population, LR led to poorer OS and CSS rates at 12 years compared with local control (LC) (71% vs. 81% [P = 0.001] and 69% vs. 88% [P < 0.001], respectively). In a Cox multiple regression model, LR was a significant predictor of disease specific mortality. The hazard ratio (HR) associated with LR was 2.69 for mortality and 2.67 for DM (P < 0.001 and P < 0.001, respectively). The median time from surgery to the development of DM was 3.8 years for patients without LR compared with 4.7 years for patients with LR. Patients who developed LR also had two peaks in the rate of DM (at 2.5 years and at 6.5 years) compared to only one peak (at 1.5 years) for patients who did not develop LR. The impact of LR on DM still was evident in patients with small tumors (< or = 2.0 cm; P < 0.001), negative lymph nodes (P = 0.004), or both (P < 0.001). Recurrent disease that developed outside of the surgical bed region had no negative effect on survival. In the matched-pair analysis (controlling for age, tumor size, grade, number of positive lymph nodes, and estrogen receptor status), LR remained the most significant predictor of mortality (HR: mortality, 5.86; DM, 6.43). CONCLUSIONS: The current results suggest that LR may be responsible for an increase in DM and disease specific mortality in patients who undergo CS and receive RT. This suggestion is reinforced by the distinct difference seen in the time distribution of DM after LR developed and by the fact that recurrent disease that originated outside of the surgical bed did not affect OS. These data reinforce the necessity to insure optimal LC in patients who are treated with breast-conserving therapy. PMID- 12569591 TI - Electronic support groups for breast carcinoma: a clinical trial of effectiveness. AB - BACKGROUND: A recent Pew Charitable Trust study found that 52,000,000 individuals used the Internet to obtain health/medical information. Clinical trials of face to-face breast carcinoma support groups show evidence of 1) improvement in quality of life, 2) reduction of psychologic symptoms, 3) improvement in coping responses, and 4) a reduction in pain. To the authors' knowledge, a few studies published to date have investigated Internet-delivered electronic support groups (ESGs) for cancer. The most sophisticated is the Comprehensive Health Enhancement Support System (CHESS) program, which provides integrated information, referral, and a newsgroup-based social support program. However, to the authors' knowledge, no studies published to date have examined the impact of a breast carcinoma ESG in a clinical trial. METHODS: Sixty-seven women completed the initial baseline questionnaires, 32 of whom accepted the authors' invitation and began the groups. With regard to geographic location, 49% lived in rural/small towns, 41% lived in medium-sized cities, and 10% lived in large cities. Diagnostic stages of disease were: Stage I, 22%; Stage II, 56%; Stage III, 12%; and other forms, 10%. There were 4 intervention groups, of which 8 participants led by trained Wellness Community (TWC) (a national agency) leaders met for 1.5 hours once a week for 16 weeks. Student t tests for paired outcome data were computed using baseline and postgroup scores. RESULTS: The results of the current study indicated that breast carcinoma patients significantly reduced depression (Center for Epidemiologic Studies-Depression [CES-D] scale) and Reactions to Pain. They also demonstrated a trend toward increases on The Posttraumatic Growth Inventory (PTGI) in two subscales: New Possibilities and Spirituality. Counterintuitively, breast carcinoma patients appeared to demonstrate an increase in emotional suppression. Postinterview results indicated that approximately 67% of patients found the group to be beneficial. Those who withdrew from the groups (20%) demonstrated low scores in their ability to contain anxiety and appeared to be more likely to suppress their thoughts and feelings regarding their illness. CONCLUSIONS: The findings of the current study are encouraging, particularly because it was conducted through TWC, a national agency willing to make this type of intervention readily available at no cost. A limitation of the current study was the lack of randomization and a control group comparison. Although the authors were not able to demonstrate effectiveness without the addition of a control condition, the analysis of pregroup and postgroup outcomes suggests that a randomized trial is worthwhile. Women with a devastating disease will join and commit themselves to an online support group. In addition, because a large percentage of these women were from rural locations, this type of intervention may hold promise for those who have limited access to support groups. PMID- 12569592 TI - Positive surgical margins and ipsilateral breast tumor recurrence predict disease specific survival after breast-conserving therapy. AB - BACKGROUND: The current study identified determinants of systemic recurrence and disease-specific survival (DSS) in patients with early-stage breast carcinoma treated with breast-conserving surgery and radiation therapy (breast-conserving therapy, or BCT). METHODS: The study population consisted of 1,043 consecutive women with Stages I or II breast carcinoma who underwent BCT between 1970 and 1994. Clinical and pathologic characteristics evaluated included age, tumor size, tumor grade, estrogen and progesterone receptor status, surgical margins, axillary lymph node involvement, and use of adjuvant therapy. RESULTS: At a median follow-up time of 8.4 years, 127 patients (12%) had developed an ipsilateral breast tumor recurrence (IBTR), and 184 patients (18%) had developed a systemic recurrence. On multivariate logistic regression analysis, tumor size greater than 2 cm, positive lymph nodes, lack of adjuvant tamoxifen therapy, and positive margins (odds ratio [OR], 3.7; 95% confidence interval [CI], 1.1-12.3; P = 0.034) were predictors of systemic recurrence. When IBTR was added into the model, adjuvant therapy and surgical margins were not independent predictors; however, IBTR was an independent predictor of systemic recurrence (IBTR vs. no IBTR; OR, 6.2; 95% CI, 3.1-12.3; P < 0.001). The 10 year DSS rate after BCT was 87%. On multivariate Cox proportional hazards model analysis, the following factors were independent predictors of poor DSS: tumor size greater than 2 cm (vs. < or = 2 cm; relative risk [RR], 2.3; 95% CI, 1.2-4.3; P = 0.010), negative progesterone receptor status (vs. positive; RR, 2.7; 95% CI, 1.4-5.1; P = 0.003), positive margins (vs. negative; RR, 3.9; 95% CI, 1.4-11.5; P = 0.011), and IBTR (vs. no IBTR; RR, 5.5; 95% CI, 2.8-11.0; P < 0.001). CONCLUSIONS: Positive surgical margins and IBTR are predictors of systemic recurrence and disease specific survival after BCT. Aggressive local therapy is necessary to ensure adequate surgical margins and to minimize IBTR. PMID- 12569593 TI - A 5-decade analysis of 13,715 carcinoid tumors. AB - BACKGROUND: Carcinoid tumors represent an unusual and complex disease spectrum with protean clinical manifestations. This compilation of several large United States-based databases comprising patients from 1950 to 1999 examines 13,715 carcinoid tumors and provides epidemiologic information regarding the natural history and evolution of the detection and diagnosis of this entity. METHODS: The authors evaluated 10,878 carcinoid tumors that were identified by the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute (NCI) from 1973 to 1999 in addition to 2837 carcinoid tumors that were registered previously by two earlier NCI programs. To the authors' knowledge, this represents the largest current epidemiology series addressing carcinoid tumors to date. RESULTS: Specific trends in incidence for carcinoid tumors of certain sites were identified. Among the most recently collected subset of data, sites that demonstrated the greatest incidence of carcinoids were the gastrointestinal tract (67.5%) and the bronchopulmonary system (25.3%). Within the gastrointestinal tract, most carcinoid tumors occurred in the small intestine (41.8%), rectum (27.4%), and stomach (8.7%). For all sites, age-adjusted incidence rates were highest in black males (4.48 per 100,000 population per year). Associated noncarcinoid tumors were frequent in conjunction with small intestinal (29.0%), gastric (20.5%), colonic (20.0%), and appendiceal (18.2%) carcinoids. The highest percentages of nonlocalized lesions were noted for cecal (81.5-83.2%) and pancreatic (71.9-81.3%) carcinoids, whereas the highest percentage of localized disease was found among rectal (81.7%), gastric (67.5%), and bronchopulmonary (65.4%) carcinoids. The best 5-year survival rates were recorded for patients with rectal (88.3%), bronchopulmonary (73.5%), and appendiceal (71.0%) carcinoids; these tumors exhibited invasive growth or metastatic spread in 3.9%, 27.5%, and 38.8% of patients, respectively. CONCLUSIONS: Carcinoids appear to have increased in overall incidence over the past 30 years; for some sites, this trend has been evident for nearly half a century. Recent marked increases in gastric and rectal carcinoids and a concomitant decrease in appendiceal carcinoid incidence may be due in part to varying rules of registration among the compiled databases examined in this report or to improvements in diagnostic technology; increased awareness of and about carcinoid tumors also may play a significant role. In 12.9% of all patients with carcinoid, distant metastases already were evident at the time of diagnosis; the overall 5-year survival rate for all carcinoid tumors, regardless of site, was 67.2%. These findings bring into question the widely promulgated relative benignity of carcinoid disease. Certain carcinoid tumors, such as those of the rectum, appear to be over-represented among the black and Asian populations within the United States, suggesting the role of genetics in the development of this intriguing disease. PMID- 12569594 TI - Vascular endothelial growth factor, CD68, and epidermal growth factor receptor expression and survival in patients with Stage II and Stage III colon carcinoma: a role for the host response in prognosis. AB - BACKGROUND: The elucidation of new therapeutic targets of prognostic significance in colon carcinoma is necessary to improve outcomes. In the current study, the authors examined the expression of epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) in primary colon carcinoma cases and VEGF in tumor-associated macrophages (TAM)/stroma, and their correlation with survival. METHODS: The authors identified 131 consecutive American Joint Committee on Cancer Stage II and Stage III colon carcinoma patients seen at the University of Rochester between 1990-1995. Expression of VEGF, EGFR, and CD68 were examined by immunohistochemistry in paraffin-embedded primary colon tumors and graded as the percentage of cells stained. Data were analyzed using a multivariate Cox proportional hazards model. RESULTS: VEGF expression in tumor was not found to be significantly associated with survival. However, 42% of the patients expressed VEGF in TAM/stroma. The median survival in this group was 9.7 years versus 4.3 years in the VEGF-negative (TAM/stroma) group (hazards ratio of 0.57, 95% confidence interval [95% CI], 0.34-0.95; P = 0.03). Although TAM infiltration alone was not found to be significant in multivariate analysis, the presence of both CD68 and VEGF (TAM/stroma) was predictive of improved survival (hazards ratio of 0.48, 95% CI, 0.28-0.83; P = 0.006). High grades of EGFR expression (> or = Grade 2) were found to be associated with a trend toward worsened survival. CONCLUSIONS: The greater than twofold increase in median survival associated with VEGF-expressing TAM suggests a hitherto unknown role for this subset of cells in the host response to colon carcinoma and requires further investigation. Overexpression of EGFR may be associated with worsened survival, providing a rationale for trials of anti-EGFR agents as adjuvant therapy. PMID- 12569595 TI - Quantitative biopsy pathology for the prediction of pathologically organ-confined prostate carcinoma: a multiinstitutional validation study. AB - BACKGROUND: Quantitative biopsy pathology with prostate specific antigen significantly improves the prediction of pathologic stage in patients with clinically localized prostate carcinoma (PCa). The authors recently reported a computational model for predicting patient specific likelihood of organ confinement of PCa using biopsy pathology and clinical data. The current study validates the initial models and presents an new, improved tool for clinical decision making. METHODS: The authors assessed 10 biopsy pathologic parameters and 2 clinical parameters using data from two institutions. Of 1287 patients, 798 men had pathologically organ confined (OC) PCa, 282 men had nonorgan-confined disease with capsular penetration (NOC-CP) only, and 207 men showed seminal vesicle or lymph node invasion (NOC-AD) after undergoing pelvic lymphadenectomy and radical prostatectomy. Patient input data were evaluated by ordinal logistic (OLOGIT) and neural network (NN) models; and the likelihood of developing OC, NOC CP, or NOC-AD disease was calculated for the combined and separate data sets and was compared with the results from original presentation. In addition, a new two output model was constructed (OC/NOC-CP vs. NOC-AD). RESULTS: The three-output OLOGIT and NN models predicted OC disease with 95.0% and 98.6% accuracy, respectively, for the combined data set and with 93.0% and 98.6% accuracy, respectively, on subset analysis. The combined accuracy for predicting OC, NOC CP, and NOC-AD disease in the entire validation set was 66.7% for OLOGIT model and 66.0% for the NN model. The two-output OLOGIT and NN models correctly predicted 94.9% and 100.0% of all OC/NOC-CP disease, respectively. CONCLUSIONS: Both computation models predicted OC PCa with an accuracy of 93.0-98.6% when they were validated with two different data sets. The OLOGIT and NN-based, two-output model permitted an appropriate treatment decision for 85.2-90.2% of patients. These data support the use of quantitative pathology and clinical data-based decision modeling to manage patients with clinically localized PCa. PMID- 12569596 TI - Long-term outcomes after treatment with external beam radiation therapy and palladium 103 for patients with higher risk prostate carcinoma: influence of prostatic acid phosphatase. AB - BACKGROUND: The objective of this study was to define the long-term prognostic significance of prostatic acid phosphatase (PAP) levels in patients with higher risk, early-stage prostate carcinoma. METHODS: One hundred sixty-one consecutive patients with Stage T1-T3 prostate carcinoma (according to the 1992 criteria of the American Joint Committee on Cancer) were treated from 1992 through 1996. Each patient had a Gleason score > or = 7 and/or a prostate specific antigen (PSA) level > 10 ng/mL. The original biopsy slides for 130 of 161 patients were retrieved and rereviewed by a single pathologist (L.T.). Enzymatic PAP measurements were determined using a standard method. Values up to 2.5 Units were considered normal. Patients received 41 grays (Gy) of external beam radiation therapy to a limited pelvic field followed 4 weeks later by a palladium 103 (Pd 103) boost using transrectal ultrasound and fluoroscopic guidance as described previously. The prescribed minimum Pd-103 dose to the prostate was 80 Gy (pre National Institute of Standards and Technology [NIST]-99). Freedom from biochemical failure was defined as a serum PSA level < or =0.2 ng/mL at last follow-up. RESULTS: There was little correlation between pretreatment PSA levels, Gleason scores, and PAP measurements. Thirty-eight patients developed biochemical failure. The overall actuarial freedom from biochemical progression at 10 years is 79%, with 118 patients followed for > 5 years. In a multivariate Cox proportional hazards analysis that considered each factor as a continuous variable, the strongest predictor of failure was PAP (P = 0.0001), followed by Gleason score (P = 0.13), and PSA (P = 0.04). PAP was especially helpful in stratifying patients with pretreatment PSA levels between 4 ng/mL and 20 ng/mL, for whom the prognosis does not different when they are subdivided into PSA categories. When the PAP subgroup analysis was limited to this relatively favorable group, there was a wide range of prognoses. CONCLUSIONS: The biochemical cure rate was remarkably high among the 161 patients evaluated. The fact that the PAP was the strongest predictor of long-term biochemical failure in patients with otherwise higher risk features reported here suggests that it may be a more accurate indicator of micrometastatic disease compared with the Gleason score and the PSA level. This report adds to the rationale for reintroducing PAP measurement into general practice. PMID- 12569597 TI - Syndromic aspects of testicular carcinoma. AB - BACKGROUND: In patients with hereditary or constitutional chromosomal anomalies, testicular carcinoma can develop sporadically or on the basis of an underlying hereditary genetic defect. Greater knowledge of these genetic defects would provide more insight into the molecular pathways that lead to testicular carcinoma. To the authors' knowledge, little attention has been paid to date to the comorbid occurrence of testicular carcinoma in patients with hereditary disorders or constitutional chromosomal anomalies. METHODS: The authors performed a review of the literature. RESULTS: Twenty-five different hereditary disorders or constitutional chromosomal anomalies have been reported in patients who developed seminomatous or nonseminomatous testicular carcinoma. CONCLUSIONS: Although most of these malignancies were too rare to enable the detection of statistically significant correlations between the chromosomal/hereditary disorder and the testicular tumor, it was striking that many of the patients had also other urogenital abnormalities. Susceptibility to urogenital abnormalities seems to disrupt normal urogenital differentiation and suggests a correlation with testicular dysgenesis and, thus, also with testicular carcinoma. Other evidence of causal involvement has been found in the field of tumor cytogenetics. Some of the genes responsible for hereditary disorders have been mapped to regions that are of interest in the development of sporadic testicular carcinoma. Molecular studies on candidate genes will be required to provide definite answers. Completion of the human gene map and the availability of advanced gene arrays and bioinformatics are expected to greatly facilitate further exploration of the role of hereditary genetic defects in testicular carcinoma. PMID- 12569598 TI - Evaluation of an instrument to assess the needs of men diagnosed with prostate carcinoma: an assessment of the validity and reliability of a self-administered questionnaire developed to measure the needs experienced by men diagnosed with prostate carcinoma. AB - BACKGROUND: This study was conducted to evaluate a self-administered questionnaire developed to measure the needs experienced by men diagnosed with prostate carcinoma (the Prostate Cancer Needs Questionnaire [PCNQv1.1]). The PCNQv1.1 was constructed in two parts. Part 1 measures the needs at diagnosis and initial treatment and Part 2 measures current needs. METHODS: A random sample of 650 men diagnosed with prostate carcinoma who were being treated by a urologist of the Hunter Urology Group in Newcastle, New South Wales, Australia, were invited to participate in the study. They were sent by post the self-administered PCNQv1.1. Completed questionnaires were received from 385 men. Information provided on 100 additional men was considered ineligible. RESULTS: The principal components method of factor analysis with varimax orthogonal rotation identified eight factors with eigenvalues greater than 1, which accounted for 61.7% of the variance in Part 1 of the PCNQv1.1. Likewise, six factors were identified in Part 2, which accounted for 63.5% of the variance. Internal reliability coefficients (Cronbach alpha) were adequate for identified factors with values ranging from 0.70 to 0.88 for Part 1 and from 0.71 to 0.87 for Part 2. Test-retest reliability was acceptable with intraclass correlation coefficients ranging from 0.61 to 0.78 for Part 1 and from 0.60 to 0.82 for Part 2. CONCLUSIONS: These results suggest that the PCNQv1.1 is a valid and reliable instrument to assess the needs experienced by men diagnosed with prostate carcinoma. PMID- 12569599 TI - Hypermethylation in promoter region of E-cadherin gene is associated with tumor dedifferention and myometrial invasion in endometrial carcinoma. AB - BACKGROUND: Loss of E-cadherin expression is associated with aberrant 5' CpG island methylation in various tumors. METHODS: The authors analyzed the methylation status and immunohistochemical expression of E-cadherin in 142 endometrial tissues, consisting of 21 normal endometria, 17 endometrial hyperplasias, and 104 endometrial carcinomas. RESULTS: All normal endometria and endometrial hyperplasias showed positive staining of E-cadherin, and methylation of the E-cadherin gene was not detected in any samples. In endometrial carcinoma, the positive ratio of methylation was higher and was associated with tumor dedifferention and myometrial invasion. In G1 endometrial adenocarcinomas, 66.7% showed positive staining and 33.3% showed heterogeneous staining. Methylation of the E-cadherin gene was detected in 15.6%. In G2 tumors, 19.0% showed positive staining, 69.0% showed heterogeneous staining and 11.9% showed negative staining. Methylation of the E-cadherin gene was found in 50.0%. In G3 tumors, 9.1% showed positive staining, 54.5% showed heterogeneous staining and 36.3% showed negative staining. Methylation of the E-cadherin gene was found in 81.8% of the tumors. Of the samples with no-myometrial invasion, 23.1% had methylation. In those with invasion in less than half of the myometrium, 28.6% did and in those with invasion of half or more of the myometrium, 55.6% had methylation. Of samples that did not have lymph node metastasis, 33.7% had methylation, whereas of samples that had lymph node metastasis, 60.0% had methylation. CONCLUSIONS: This is the first report to analyze methylation of the E-cadherin gene promoter of endometrial carcinoma and the evidence suggests that methylation of the E cadherin gene occurs in association with the acquisition of invasive capacity. PMID- 12569600 TI - Characterization of laryngopharyngeal reflux in patients with premalignant or early carcinomas of the larynx. AB - BACKGROUND: An association between laryngopharyngeal reflux (LPR) and laryngeal carcinoma has been suggested, but remains unproven. The current pilot study was performed to determine the incidence of LPR among patients with early laryngeal carcinomas or dysplasia and to examine the associations between levels of LPR and histologic stage, smoking status, the symptom of heartburn, and body position during reflux episodes. METHODS: Behavioral and 24 hour pH monitoring data were prospectively acquired and analyzed for 40 previously untreated adults with dysplasia, T1 or T2 laryngeal carcinomas. RESULTS: Eighty-five percent of patients had LPR. No significant association was shown between the level of LPR and histologic stage or smoking status. Heartburn did not predict LPR. The incidence of LPR in the upright body position was 91%, compared with 9% in the supine position, among patients with LPR. CONCLUSIONS: When compared to available normative data, the current findings show a high incidence of LPR in patients with premalignant and early laryngeal cancer. These findings highlight the need for a matched-control study evaluating LPR as a potential predisposing factor for laryngeal carcinoma. PMID- 12569601 TI - Overexpression of Glut-1 and increased glucose metabolism in tumors are associated with a poor prognosis in patients with oral squamous cell carcinoma. AB - BACKGROUND: The overexpression of glucose transporters, especially of Glut-1, is a common characteristic of human malignancies, including head and neck carcinoma. Recently, the assessment of glucose metabolism in the tumor with [(18)F]-2-fluoro 2 deoxy-D-glucose (FDG) and positron emission tomography (FDG-PET) has been used to identify particularly aggressive tumors. The authors tested the hypothesis that both glucose transport and its metabolism play a key role in the progression of oral squamous cell carcinoma (OSCC). METHODS: Retrospective analysis of Glut-1 expression was performed by immunohistology in 118 patients with OSCC, and a Glut 1 labeling index (LI) was established for each. A separate group of 44 patients with primary OSCC was evaluated prospectively by FDG-PET prior to surgery. To link the expression of Glut-1 with glucose metabolism, both FDG-PET and immunohistology were determined in a subgroup of 31 patients, and the results were correlated with overall survival. RESULTS: The patients who had OSCC with a low LI for Glut-1 survived significantly longer compared with patients who had OSCC with a high LI (138 months vs. 60 months; P = 0.0034). It was found that Glut-1 expression was an independent marker of prognosis in patients with OSCC. In patients who were evaluated by FDG-PET, the standardized uptake value (SUV) below the median split value of 5.6 was predictive of a longer survival (P < 0.027), whereas an SUV > 5.6 was associated with an increased hazard of death. In combination, a high Glut-1 level and a high SUV predicted shorter survival (P < 0.005) for patients with OSCC. Patients who achieved a complete response to preoperative radiation tended to have tumors with low glucose metabolism, as defined by both the Glut-1 LI and the SUV. CONCLUSIONS: Both glucose transport and glucose metabolism determine the glycolytic tumor phenotype, which is a significant negative biomarker of prognosis and overall survival in patients with OSCC. PMID- 12569602 TI - Refractory Aspergillus pneumonia in patients with acute leukemia: successful therapy with combination caspofungin and liposomal amphotericin. AB - BACKGROUND: Pulmonary aspergillosis and other invasive fungal infections (IFIs) commonly complicate the management of patients with acute leukemia. Standard amphotericin-based therapies may be ineffective for many patients and the available salvage agents (itraconazole and caspofungin) are reported to possess only moderate activity against resistant infections. Laboratory evidence suggests a synergistic interaction between amphotericin and caspofungin. The authors treated a group of patients with amphotericin-refractory IFIs with the combination of caspofungin and amphotericin (or liposomal amphotericin). METHODS: A retrospective evaluation of patients with amphotericin-resistant IFIs was conducted. Diagnosis was based on clinical, radiographic, and when available, microbiologic data. Response to combination antifungal therapy was graded as either favorable or unfavorable. Favorable responses included improvement of both clinical and radiographic signs of fungal pneumonia. All other responses were graded as unfavorable. RESULTS: Thirty patients were included in this analysis. Twenty-six patients had acute leukemia. Based on recently published criteria, the IFIs were classified as proven in 6 patients, probable in 4 patients, and possible in 20 patients. The median duration and dose of amphotericin monotherapy were 12 days (range, 4-65 days) and 7.8 mg/kg (range, 4.2-66.1 mg/kg), respectively. The median duration of combination therapy was 24 days (range, 3-74 days). Eighteen patients (60%) experienced a favorable antifungal response. Twenty patients with acute leukemia received combination therapy for fungal pneumonias arising during intensive chemotherapy treatments. Favorable responses were observed in 15 of these patients (75%), and antifungal response did not depend on the response of the underlying leukemia. Survival to hospital discharge was significantly better (P < 0.001) in patients having a favorable response. Mild to moderate nephrotoxicity was noted in 50% of patients, necessitating the substitution of liposomal amphotericin. Mild elevation of alkaline phosphatase levels occurred in 30% of patients. Caspofungin was temporarily withheld from one patient who developed moderate but reversible biochemical hepatotoxicity. CONCLUSIONS: The antifungal combination of caspofungin and amphotericin can be administered safely to high-risk patients with hematologic malignancies. Although an absolute assessment of efficacy is limited by the design of this study, encouraging outcomes were noted for many patients. The authors plan to evaluate this regimen further in a randomized clinical trial. PMID- 12569603 TI - Complete cytogenetic and molecular responses to interferon-alpha-based therapy for chronic myelogenous leukemia are associated with excellent long-term prognosis. AB - BACKGROUND: Little is known regarding long-term prognosis among patients with Philadelphia chromosome (Ph)-positive chronic myelogenous leukemia (CML) who achieve a complete cytogenetic response (0% Ph-positive cells) after treatment with interferon-alpha. METHODS: The authors analyzed 512 patients with Ph positive, early chronic-phase CML who were treated with interferon-based therapies between 1981-1995 for the incidence and durability of complete cytogenetic response, and in relation to long-term prognosis. RESULTS: One hundred forty patients (27%) achieved a complete cytogenetic response. Their 10 year survival rate was 78%. At the time of last follow-up, 44 patients (31%, 9% of the total) were alive, 21 in first and 23 in second durable complete cytogenetic response (median, 127 months; range, 88-191 months); 39 patients had not received any therapy for a median of 50 months (range, 11-139 months). Analysis by reverse transcriptase-polymerase chain reaction in 78 patients during complete cytogenetic response showed 46 who had achieved at least 1 complete molecular response. Five of these 78 patients had died by the time of last follow up, but only 2 had died of disease-specific causes. Recurrence rates were significantly lower and cytogenetic response durations were significantly longer among patients who achieved at least one complete molecular response. CONCLUSIONS: Achieving a complete cytogenetic or molecular response after therapy with interferon-alpha appears to be associated with excellent long-term prognosis. Approximately 10% of patients reportedly can achieve durable complete cytogenetic response, with or without continuation of interferon. This finding emphasizes the potential of long-term event-free survival in CML patients outside the context of allogeneic stem cell transplantation, which may be improved with new therapies such as imatinib mesylate. PMID- 12569604 TI - Evaluation of responses to chemoembolization in patients with unresectable hepatocellular carcinoma. AB - BACKGROUND: The authors used computed tomography (CT) scans to correlate the changes in tumor vascularity, necrosis, and size with response and survival after transcatheter arterial chemoembolization (TACE) in patients with advanced, unresectable, hepatocellular carcinoma (HCC). METHODS: The authors studied 72 patients with biopsy-proven, unresectable HCC and focused on 186 individual tumor masses. A baseline, multiphase, helical CT was performed and at least three follow-up CT scans were performed after treatment by TACE. Tumors were classified as hypervascular or hypovascular and patients were classified as responders or nonresponders based on CT evidence of altered tumor size, tumor necrosis, and the appearance of new tumors. A new scoring system was used to monitor patient response to TACE. RESULTS: Thirty-eight patients were responders and 34 were nonresponders. Patient survival was significantly increased (P = 0.009) in patients who were hypervascular responders. Survival also was increased in hypervascular nonresponders compared with hypovascular nonresponders (P = 0.008) and in hypovascular responders compared with hypovascular nonresponders (P = 0.002). Response to chemoembolization was found to be significantly (P = 0.02) and inversely proportional to tumor size, but the number of tumor foci in an individual patient was not predictive. CONCLUSIONS: TACE appears to result in improved survival among HCC patients with hypervascular tumors who responded to therapy. However, even patients classified by CT as hypervascular nonresponders and hypovascular responders have improved survival. PMID- 12569605 TI - Angiographic subsegmentectomy for the treatment of patients with small hepatocellular carcinoma. AB - BACKGROUND: The therapeutic results of nonsurgical treatment for patients with hepatocellular carcinoma (HCC) have been poor, and improved treatments are needed. The authors recently developed a new technique called angiographic subsegmentectomy for the treatment of patients with small HCC. METHODS: The technique includes confirming the diagnosis of small HCC using a helical computed tomography (CT) scan combined with an angiography system for identifying the tumor-feeding subsegmental hepatic artery, injecting lipiodol containing farmorubicin until it enters the portal vein in sufficient amounts, and injecting sponge particles into the hepatic artery for embolization. Occlusion of the hepatic artery with gel particles and occlusion of the portal vein by lipiodol induce infarction necrosis, which encompasses the entire tumor and the surrounding liver parenchyma. RESULTS: The treatment was given to 23 patients with 30 HCC tumors that measured < 20 mm in greatest dimension. It was successful in all 23 patients. Serum alanine aminotransferase levels were elevated to a significant level in the majority of patients after treatment, mild ascites developed in three patients, and the patients complained of pain and fever posttreatment that were controlled readily. No patients developed hepatic failure. Only one patient developed recurrent disease posttreatment at 1.5 years, for a recurrence rate of 5% at 1 year and 6.6% at 1.5 years, a rate that has never been achieved with other treatment modalities. CONCLUSIONS: Angiographic subsegmentectomy is a novel treatment for patients with small HCC. The results indicated that it is equivalent to undergoing small resection and is superior to conventional arterial chemoembolization. PMID- 12569606 TI - Detection of tumor specific gene expression in bone marrow and peripheral blood from patients with small cell lung carcinoma. AB - BACKGROUND: Small cell lung carcinoma (SCLC) has the propensity to grow rapidly and metastasize extensively. Detection of micro-dissemination of SCLC may have clinical relevance. For its detection, tumor-specific gene expressions were examined in peripheral blood and bone marrow aspirate from patients with SCLC. METHODS: Expression of prepro-gastrin-releasing peptide (preproGRP), neuromedin B receptor (NMB-R) and gastrin-releasing peptide receptor (GRP-R) were examined by reverse transcriptase polymerase chain reaction (RT-PCR) in peripheral blood and bone marrow aspirate from 40 untreated patients with SCLC. Control samples consisted of peripheral blood samples from 5 patients with nonsmall cell lung cancer (NSCLC) and 20 healthy volunteers. RESULTS: Positive rates of preproGRP, NMB-R, and GRP-R in bone marrow aspirate of patients with SCLC were 23% (9/40), 8% (3/40), and 10% (4/40), respectively. Those rates in peripheral blood were 11% (4/38), 5% (2/38), and 29% (11/38), respectively. Although GRP-R expression was detected in patients with NSCLC and in healthy volunteers, preproGRP and NMB-R expressions were not detected in patients with NSCLC and in healthy volunteers. All three gene expressions in bone marrow were more frequently observed in patients with bone marrow metastasis, accessed by biopsy, than in patients without. PreproGRP gene expression in bone marrow was also more frequent in patients with bone metastasis, accessed by bone scintigram, than in patients without, and was related to poorer survival. CONCLUSIONS: Micro-dissemination of SCLC was detectable by RT-PCR of preproGRP and NMB-R, both specific for SCLC. These gene expressions in bone marrow may be related to disease extent and prognosis. PMID- 12569608 TI - Primitive neuroectodermal tumors in the central nervous system following cranial irradiation: a report of four cases. AB - BACKGROUND: Radiation induced intracranial neoplasms are uncommon but well described and include gliomas, meningiomas, and sarcomas. The development of primitive neuroectodermal tumors (PNETs) following prophylactic craniospinal irradiation has been infrequently reported previously. The authors present four additional cases of PNETs that developed after previous cranial irradiation. METHODS: Four patients who had previously been irradiated were determined to have PNETs of the central nervous system characterized by histopathologic and immunohistochemical features. The average patient age at diagnosis of the initial tumors and cranial irradiation was 17 years. The PNETs developed 5, 11, 11, and 18 years after completion of radiation. RESULTS: Three patients had posterior fossa tumors, one pilocytic astrocytoma, one low grade astrocytoma, and one malignant ependymoma, which had been diagnosed and treated in childhood. Two of those patients developed supratentorial PNETs and the third a cerebellar hemispheric PNET. The fourth patient developed a posterior fossa PNET following irradiation for a temporal astrocytoma, which was initially diagnosed and resected at 37 years of age. Mean survival was 12 months after diagnosis. CONCLUSIONS: The development of PNETs after cranial irradiation may be more common than thought previously and should be considered in the differential diagnosis of irradiation induced neoplasms. Survival after diagnosis of these radiation induced PNETs was short, and this may reflect an inability to provide standard therapy used for primary PNETs. PMID- 12569607 TI - Prognostic implication of clinical, radiologic, and pathologic features in patients with anaplastic gliomas. AB - BACKGROUND: The clinical evolution of anaplastic glioma (anaplastic astrocytoma, oligodendroglioma, and oligoastrocytoma) is variable. Previous studies merged patients with anaplastic glioma and the much more common glioblastoma multiforme. Therefore, the conclusions on prognostic factors reflected in part the consequences of an analysis in a heterogeneous population. METHODS: To identify clinical, neuroradiologic, pathologic, and molecular factors with prognostic significance, we analyzed 95 treated patients with a histologic diagnosis of anaplastic glioma. Variables included age, gender, clinical manifestations at diagnosis (seizures, focal neurologic deficit, and cognitive changes), computed tomographic (CT) scan characteristics (diffuse, ring, and no enhancement), tumor location, extent of resection, histopathology, postoperative Karnofsky performance status (KPS) score, adjuvant chemotherapy, tumor response, proliferation index (Ki-67 expression), and p53, p16, pRb, and epidermal growth factor receptor immunohistochemical expression. RESULTS: Ninety-five patients with a histologic diagnosis of anaplastic astrocytoma (73%), anaplastic oligoastrocytoma (16.6%), or anaplastic oligodendroglioma (10.4%) constituted the basis of this study. Median overall survival was 29 months. Multivariate analysis revealed that an age of 49 years or younger (P < 0.03), postoperative KPS score of 80 or higher (P < 0.007), absence of ring enhancement (P = 0.03), and a proliferation index of 5.1% or lower (P = 0.044) were independently associated with longer survival. The presence of an oligodendroglial component was associated with better prognosis in the univariate analysis (P = 0.009), although this lost power in the multivariate analysis. CONCLUSIONS: In addition to previously recognized prognostic variables such as age and KPS score, CT ring enhancement and tumor proliferation index were identified as independent predictors of survival in a homogeneous series of patients with anaplastic gliomas. PMID- 12569609 TI - Expression of survivin in astrocytic tumors: correlation with malignant grade and prognosis. AB - BACKGROUND: Astrocytic tumors are the most common tumors of the central nervous system. The mechanisms of genetic change of astrocytic tumors have not been understood completely. Recently, survivin has been identified as a member of the inhibitor-of-apoptosis family. Survivin expression is considered an important prognostic factor of many tumors. METHODS: The authors investigated 43 astrocytic tumors (8 diffuse astrocytomas; 15 anaplastic astrocytomas; 20 glioblastomas). The authors examined survivin mRNA expression in tumor specimens by reverse transcriptase-polymerase chain reaction amplification. The authors analyzed the relationship between survivin expression and other molecular changes commonly found in astrocytic tumors [p53 alteration, overexpression of epidermal growth factor receptor (EGFR), and bcl-2 expression]. RESULTS: Thirty-four of 43 (79.1%) astrocytic tumors expressed survivin. The distributions included 3 of 8 (37.5%) diffuse astrocytomas, 13 of 15 (86.7%) anaplastic astrocytomas, and 18 of 20 (90.0%) glioblastomas. Expression of survivin (P = 0.0057) and EGFR (P = 0.0112) was significantly associated with malignant grade of astrocytic tumors, but expression of p53 (P = 0.1893) and bcl-2 (P = 0.2552) was not. Furthermore, patients with survivin-positive astrocytic tumors had significantly shorter overall survival times compared with patients who had survivin-negative tumors (P = 0.0271). CONCLUSIONS: Survivin expression in astrocytic tumors varies with histologic malignancy and may play an important role in the oncogenesis and progression of astrocytic tumors. These data suggest that survivin has great potential as a therapeutic target in astrocytic tumors. PMID- 12569610 TI - Survival and functional outcome of children with hypothalamic/chiasmatic tumors. AB - BACKGROUND: The management of children with hypothalamic (H) and/or chiasmatic (C) tumors remains controversial. We evaluated the impact of clinical and neuroimaging parameters and primary therapy on overall (OS) and progression-free (PFS) survival and on neuroendocrine and neurocognitive outcome in children with H and/or C tumors. METHODS: Records were reviewed for 73 children with H and/or C tumors treated at St. Jude Children's Research Hospital between October 1981 and December 1999. RESULTS: Thirty-six patients received irradiation or chemotherapy immediately postdiagnosis and 37 were observed. The 6-year OS and PFS rates were 86 +/- 5%; and 36 +/- 7%, respectively. The 6-year PFS rates for the irradiation, chemotherapy, and observation groups were 69 +/- 16%, 12 +/- 11%, and 37 +/- 9%, respectively. In multivariate analysis, intracranial NF1 lesions (P = 0.015) and initial irradiation (P = 0.056) led to better PFS rates. There was no difference in OS between those initially treated or observed. Mean serial intelligence quotient (IQ) scores were 86 and 86 at diagnosis and at 6 years later, respectively. Patients younger than 5 years old had a lower mean IQ score at diagnosis (79.1) than older patients (96.3; P = 0.003). Patients who were irradiated at diagnosis had a significantly higher cumulative incidence of endocrinopathy at 3 years (P = 0.008). CONCLUSIONS: Overall survival for children with H and/or C tumors is excellent. Initial treatment with radiation and the presence of intracranial NF1 lesions were positive predictors of PFS. Mean IQ is significantly compromised at diagnosis, but does not change over time or with irradiation. Overall survival is not affected by initial observation. We recommend observation in asymptomatic patients, platinum-based chemotherapy in younger patients, and irradiation in older symptomatic patients. PMID- 12569611 TI - Stereotactic linear accelerater-based radiosurgery for the treatment of patients with glomus jugulare tumors. AB - BACKGROUND: The optimal treatment for patients with glomus jugulare tumor (GJT) of the skull base remains controversial. Surgical excision is associated with a high incidence of cranial nerve injury, decreased quality of life, and high mortality. Fractionated radiotherapy is used to control the majority of these tumors, but disadvantages are a prolonged therapy interval and exposition of adjacent brain tissue to irradiation. The authors present the results of a study on 12 of 14 consecutively admitted patients who were treated using linear accelerator-based radiosurgery (LINAC-RS), an innovative method for the treatment of GJT. METHODS: From May 1991 to March 2001, 14 patients with GJT were treated with stereotactic LINAC-RS for continued growth of tumor or of remaining tumor after surgery. Twelve patients (9 women and 3 men; age range, 28-71 years; median age, 59 years) with a median follow-up of 4 years (range, 0.8-9,0 years), were selected for retrospective analysis. A median single dose of 15 grays (Gy; range, 11-20 Gy) was applied to the surface of the tumor. RESULTS: After undergoing LINAC-RS, 8 of 12 patients (67%) reported partial or complete subjective improvement, whereas complaints remained unchanged in 4 patients (33%). Neurologic status improved in 3 patients (25%) and remained unchanged in 8 patients (67%). Magnetic resonance images showed tumor shrinkage in 8 patients (67%) and no further progression in 4 patients (33%). CONCLUSIONS: LINAC-RS is an effective and safe therapy for patients with GJT and may be used as an alternative to surgical resection. Compared with fractionated radiotherapy, LINAC RS has some advantages. However, to clarify the question of long-term tumor control, longer observation times are required. PMID- 12569612 TI - Recent trends and racial/ethnic differences in the incidence and treatment of ductal carcinoma in situ of the breast in California women. AB - BACKGROUND: The rapid increase in the incidence of ductal carcinoma in situ (DCIS) of the breast in the U.S. has been associated with the widespread adoption of screening mammography. Little is known regarding the incidence and treatment of DCIS in women of racial/ethnic groups other than white and black. The current investigation examined recent trends and racial/ethnic differences in the incidence and treatment of DCIS in California. METHODS: All cases of DCIS diagnosed in women age > or = 40 years in California between 1988-1999 were included. Age-adjusted incidence rates for white, black, Hispanic, and Asian Pacific Islander women were calculated using the 2000 U.S. female population as the standard. The estimated annual percent change (EAPC) in the rates was calculated using least squares regression. RESULTS: The average annual age adjusted incidence of DCIS (1988-1999) was 45.3 per 100,000 in white women, 35.0 in black women, 30.9 in Asian-Pacific Islander women, and 21.8 in Hispanic women. Although a steady increase in the incidence of DCIS was noted in all racial/ethnic groups over the study period, Asian-Pacific Islander women were found to have experienced the steepest increase (EAPC = 9.1%), particularly in the age group 50-64 years (EAPC = 12.0%). The DCIS incidence was reported to increase with age in white, black, and Hispanic women, but remained fairly constant after the age of 50 years in Asian-Pacific Islanders. The proportion of women with DCIS treated with mastectomy decreased from 53% in 1988 to 32% in 1999. Younger women and Asian-Pacific Islander women reportedly were more likely to undergo mastectomy. CONCLUSIONS: Considerable differences by race/ethnicity and age were observed in DCIS incidence and the change in the incidence in California between 1988 and 1999. Further information is needed to determine whether these differences are because of differential utilization of screening mammography or biologic characteristics of DCIS lesions. PMID- 12569613 TI - Can older cancer patients tolerate chemotherapy? A prospective pilot study. AB - BACKGROUND: To the authors' knowledge, few data currently are available regarding the tolerance to chemotherapy in older cancer patients. This prospective pilot study evaluated the changes in functional, mental, nutritional, and comorbid status, as well as the quality of life (QOL), in geriatric oncology patients receiving chemotherapy. METHODS: Sixty patients age > or = 70 years who were undergoing cancer chemotherapy were recruited in a university-based comprehensive cancer center. Changes in physical function were measured by the Eastern Cooperative Oncology Group performance status (ECOG PS) and Instrumental Activities of Daily Living (IADLs), mental health changes were measured by the Mini-Mental State Examination and the Geriatric Depression Scale (GDS), comorbidity was measured by Charlson's index and the Cumulative Illness Rating Scale-Geriatric, nutrition was measured by the Mini-Nutritional Assessment, and QOL was measured by the Functional Assessment of Cancer Therapy-General (FACT-G). Changes were assessed at baseline and at the end of treatment (EOT). Grade 4 hematologic and Grade 3-4 nonhematologic toxicities were recorded. RESULTS: Thirty-seven patients (63%) completed both assessments. Older cancer patients demonstrated a significant decline in measurements of physical function after receiving chemotherapy, as indicated by changes in scores on the IADL (P = 0.04) and on the physical (P = 0.01) and functional (P = 0.03) subscales of the FACT-G. They also displayed worse scores on the GDS administered postchemotherapy (P < 0.01). Patients who experienced severe chemotoxicity had more significant declines in ECOG PS (P = 0.03), IADL (P = 0.03), and GDS (P = 0.04), and more gain in the social well-being subscale (P = 0.02) of the FACT-G, than those who did not experience severe chemotoxicity. However, changes in most scores were small in magnitude clinically. No significant change was found between baseline and EOT in nutrition, comorbidity, and other aspects of the FACT-G. CONCLUSIONS: Older cancer patients undergoing chemotherapy may experience toxicity but generally can tolerate it with limited impact on independence, comorbidity, and QOL levels. It is important to recognize and monitor these changes during geriatric oncology treatment. PMID- 12569614 TI - Utilization of special education services and educational attainment among long term survivors of childhood cancer: a report from the Childhood Cancer Survivor Study. AB - BACKGROUND: The objective of the current report was to compare the self-reported rates of special education (SE) and educational attainment among specific groups of childhood cancer survivors and a random sample of sibling controls. METHODS: The Childhood Cancer Survivor Study is a retrospective cohort of individuals who were diagnosed with a cancer in childhood and survived at least 5 years postdiagnosis. This analysis includes 12,430 survivors and 3410 full siblings. Reported use of SE services and educational attainment were analyzed within subgroups defined by type of cancer, age at diagnosis, and type of treatment. RESULTS: The use of SE services was reported in 23% of survivors and 8% of siblings, with the greatest differences observed among survivors who were diagnosed before age 6 years, most notably survivors of central nervous system (CNS) tumors (odds ratio [OR], 18.8; 95% confidence interval [95%CI], 15.01 23.49), leukemia (OR, 4.4; 95%CI, 3.75-5.16), and Hodgkin disease (OR, 4.4; 95%CI, 2.64-7.24). It was found that intrathecal methotrexate (IT MTX) and cranial radiation (CRT), administered alone or in combination, significantly increased the likelihood that a survivor would use SE (IT MTX only: OR, 1.3; 95%CI, 1.09-1.78; CRT only: OR, 7.2; 95%CI, 6.14-8.39; IT MTX and CRT combined: OR, 2.6; 95%CI, 2.30-2.95). A positive dose response was identified between higher doses of CRT and use of SE. It was determined that survivors of leukemia (OR, 1.6; 95%CI, 1.23-2.16), CNS tumors (OR, 2.7; 95%CI, 1.92-3.81), non-Hodgkin lymphoma (OR, 1.8; 95%CI, 1.15-2.78), and neuroblastoma (OR, 1.7; 95%CI, 1.14 2.61) were significantly less likely to finish high school compared with siblings; however, when survivors received SE services, risk estimates approximated those of the sibling SE population. CONCLUSIONS: Children who are diagnosed with cancer should be followed closely during and after treatment to identify early signs of learning disabilities and to maximize intervention strategies for the successful completion of scholastic goals. PMID- 12569615 TI - Validation of a treatment policy for patients with prostate specific antigen failure after three-dimensional conformal prostate radiation therapy. AB - BACKGROUND: The objective of this report was to present an outcomes validation for the Fox Chase Cancer Center (FCCC) management policy for patients who demonstrate prostate specific antigen (PSA) failure after receiving three dimensional conformal radiation therapy (3DCRT). METHODS: Eligible patients included 248 men with T1-T3N0M0 prostate carcinoma who demonstrated PSA failure (according to the American Society for Therapeutic Radiology and Oncology definition) after completing definitive 3DCRT alone or with androgen deprivation (AD) therapy between May 1989 and November 1997. The primary endpoint evaluated was freedom from distant metastasis (FDM). The secondary endpoints evaluated included cause specific survival (CSS) and overall survival (OS). The variables evaluated in the multivariate analyses (MVA) included initial PSA, Gleason score, T classification, dose, PSA nadir, time to PSA failure, PSA doubling time (PSADT), initial use of AD therapy, and the use of AD therapy upon PSA failure. RESULTS: The 5-year FDM, CSS, and OS rates for the entire group were 76%, 92%, and 76%, respectively. It was found that four variables were independent predictors of FDM: Gleason score (P = 0.0039), PSA nadir (P = 0.0001), PSADT (P = 0.0001), and the use of AD on PSA failure (P = 0.0001). One hundred forty-eight men demonstrated a PSADT < 12 months. AD therapy was started in 59 men, and 89 men refused AD therapy and were observed. The use of AD therapy was associated with a significant improvement in the 5-year FDM rate (57% vs. 78%; P = 0.0026). In the group of men with PSADT < 12 months, the median time to distant failure was significantly longer in the men who received AD therapy (6 months vs. 25 months; P = 0.02). Of the 100 men with a PSADT > or = 12 months, 89 men were observed, and 11 men received AD therapy. There was no improvement in the 5-year FDM rate with the use of AD therapy compared with observation (88% vs. 92%, respectively; P = 0.74). CONCLUSIONS: The current results validate the use of PSADT as an indicator of patients who may be observed expectantly or treated with AD therapy for PSA failure after 3DCRT. Prospective trials are needed to define further the optimal treatment for these patients. PMID- 12569616 TI - Combination chemotherapy for desmoid tumors. AB - BACKGROUND: Desmoid tumor (aggressive fibromatosis) is an aggressive fibroblastic proliferation of well circumscribed, locally invasive, differentiated fibrous tissue. For patients with desmoid tumors that are not amenable to surgery or radiation therapy, the use of hormonal agents and nonsteroidal antiinflammatory drugs (NSAIDs) have been attempted, with some success. The use of chemotherapy also has been reported to have activity. METHODS: Seven patients (5 males and 2 females) with a median age of 40 years (range, 17-66 years) who received cytotoxic chemotherapy (combinations of cyclophosphamide and doxorubicin; mitomycin, doxorubicin, and cisplatin; and ifosfamide and etoposide) for desmoid tumor were reviewed retrospectively. Five patients were found to have recurrent tumors. Four patients had familial adenomatous polyposis. Four patients had failed tamoxifen and six had failed NSAIDs prior to receiving cytotoxic chemotherapy. In six patients the desmoid tumor was intraabdominal and one tumor had occurred on the buttock. RESULTS: Patients received a median number of six cycles of chemotherapy (range, two to eight cycles). Objective disease regression occurred in 3 patients. There was an apparent clinical benefit in six patients with the duration of benefit ranging from 3 months to 15 years. The chemotherapy was well tolerated and no treatment-related mortality was reported. CONCLUSIONS: The results of the current study indicate that the use of combination chemotherapy for desmoid tumors may provide long-term clinical benefits. PMID- 12569619 TI - Development of a novel process for the biological conversion of H2S and methanethiol to elemental sulfur. AB - The feasibility of anaerobic treatment of wastewater containing methanethiol (MT), an extremely volatile and malodorous sulfur compound, was investigated in lab-scale bioreactors. Inoculum biomass originating from full-scale anaerobic wastewater treatment facilities was used. Several sludges, tested for their ability to degrade MT, revealed the presence of organisms capable of metabolizing MT as their sole source of energy. Furthermore, batch tests were executed to gain a better understanding of the inhibition potential of MT. It was found that increasing MT concentrations affected acetotrophic organisms more dramatically than methylotrophic organisms. Continuous reactor experiments, using two lab scale upflow anaerobic sludge bed (UASB) reactors (R1 and R2), aimed to determine the maximal MT load and the effect of elevated sulfide concentrations on MT conversion. Both reactors were operated at a hydraulic retention time (HRT) of about 7 hours, a temperature of 30 degrees C, and a pH of between 7.3 and 7.6. At the highest influent MT concentration applied, 14 mM in R1, corresponding to a volumetric loading rate of about 50 mM MT per day, 87% of the organic sulfur was recovered as hydrogen sulfide (12.2 mM) and the remainder as volatile organic sulfur compounds (VOSCs). Upon decreasing the HRT to 3.5 to 4.0 h at a constant MT loading rate, the sulfide concentration in the reactor decreased to 8 mM and MT conversion efficiency increased to values near 100%. MT conversion was apparently inhibited by the high sulfide concentrations in the reactor. The specific MT degradation rate, as determined after 120 days of operation in R1, was 2.83 +/- 0.27 mmol MT g VSS(-1) day(-1). During biological desulfurization of liquid hydrocarbon phases, such as with liquefied petroleum gas (LPG), the combined removal of hydrogen sulfide and MT is desired. In R2, the simultaneous addition of sodium sulfide and MT was therefore studied and the effect of elevated sulfide concentrations was investigated. The addition of sodium sulfide resulted in enhanced disintegration of sludge granules, causing significant washout of biomass. Additional acetate, added to stimulate growth of methanogenic bacteria to promote granulation, was hardly converted at the termination of the experimental period. PMID- 12569620 TI - Modeling O2 transport within engineered hepatic devices. AB - Predicting and improving oxygen transport within bioartificial liver (BAL) devices continues to be an important engineering challenge since oxygen is one of the critical nutrients necessary for maintaining hepatocyte viability and function. Such a computational model would not only help predict outcomes but it would also allow system modifications to be analyzed prior to developing experimental protocols. This would help to facilitate future design improvements while reducing both experimental time and capital resource costs, and is the focus of the current study. Specifically, a computational model of O(2) transport through collagen and microporous collagen ECMs is analyzed for hollow fiber (HF), flat plate (FP), and spheroid BAL designs. By modifying the O(2) boundary conditions, hepatocyte O(2) consumption levels, O(2) permeability of the ECM, and ECM void fractions, O(2) transport predictions are determined for each system as a function of time and distance. Accuracy of the predictive model is confirmed by comparing computational vs. experimental results for the HF BAL system. The model's results indicate that O(2) transport within all three BAL designs can be improved significantly by incorporating the enhancement technique. This technique modifies a diffusion-dominant gel ECM into a porous matrix with diffusive and convective flows that mutually transport O(2) through the ECMs. Although tortuous pathways increase the porous ECM's overall effective length of O(2) travel, the decreased transport resistances of these pathways allow O(2) to permeate more effectively into the ECMs. Furthermore, because the HF design employs convective flow on both its inner and outer ECM surfaces, greater control of O(2) transport through its ECM is predicted, as compared with the single O(2) source inputs of the flat plate and spheroid systems. The importance of this control is evaluated by showing how modifying the O(2) concentration and/or transfer coefficients of the convective flows can affect O(2) transport. PMID- 12569621 TI - Theory of pH-stat titration. AB - Innovative techniques are being studied to assess the activity of bioreactors and to improve the performance and operational stability of biological processes. Among these techniques, the pH-stat titration is applicable to any bioreaction involving pH variations. Up to now, the main application of the pH-stat titration has been for nitrification monitoring. In this article, we present a theoretical model of pH-stat titration, which predicts the response to any reaction involving the production or consumption of protons, hydroxyl ions, or inorganic carbon chemical species (CO(2), HCO(3)(-), CO(3)(=)). This model is a useful tool to understand pH-stat titrations, to define their applicability and limits, and to select the best experimental conditions for specific applications. Tests have been performed to compare experimental pH-stat titration rates in the presence of carbon dioxide and HCO(3)(-) producing reactions to the values predicted by the model and a very satisfying correspondence was found. PMID- 12569622 TI - Inhibition of an activated Ras protein with genetically selected peptide aptamers. AB - Mutant alleles of Ras maintain an activated, GTP-bound conformation and relay mitogenic signals that cannot be turned off. A genetic selection in Saccharomyces cerevisiae was used to identify peptide aptamers that suppress the growth arrest phenotype of an activated Ras allele. Peptide aptamers were expressed as C terminal fusions to glutathione-S-transferase. Modifications that alter the coding capacity of the peptide aptamer indicate it is necessary for Ras2-Val19 suppression. Aptamer expression also reduces the elevated levels of cAMP and suppresses the heat shock sensitivity characteristic of Ras-activated yeast cells. The peptide aptamer retains suppressor activity when fused to thioredoxin. The peptide aptamer expression strategy described here indicates that aptamers presented as unconstrained peptides have functional capacity in vivo. PMID- 12569623 TI - A reversible hydrogel membrane for controlling the delivery of macromolecules. AB - Glucose-sensitive hydrogel membranes have been synthesized and characterized for their rate-of-delivery of macromolecules. The mechanism for changing this rate is based on variable displacement of the affinity interaction between dextran and concanavalin A (con A). Our main objective was to characterize the diffusion of model proteins (insulin, lysozyme, and BSA) through the membrane, in response to changes in environmental glucose concentrations. Membranes were constructed from crosslinked dextrans to which con A was coupled via a spacer arm. Changes in the porosity of the resulting hydrogel in the presence of glucose led to changes in the diffusion rate observed for a range of proteins. Gels of specified thickness were cast around to nylon gauze support (pore size, 0.1 mm) to improve mechanical strength. Diffusion of proteins through the gel membrane was determined using a twin-chamber diffusion cell with the concentrations being continuously monitored using a UV-spectrophotometer. Changes in the transport properties of the membranes in response to glucose were explored and it was found that, while 0.1M D-glucose caused a substantial, but saturateable, increase in the rates of diffusion of both insulin and lysozyme, controls using glycerol or L-glucose (0.1M) had no significant effect. Sequential addition and removal of external glucose in a stepwise manner showed that permeability changes were reversible. As expected, diffusion rates were inversely proportional to membrane thickness. A maximum increase in permeability was observed at pH 7.4 and at 37 degrees C. The results demonstrate that this hydrogel membrane functions as a smart material allowing control of solute delivery in response to specific changes in its external environment. PMID- 12569624 TI - A new VFA sensor technique for anaerobic reactor systems. AB - A key parameter for understanding and controlling the anaerobic biogas process is the concentration of volatile fatty acids (VFA). However, this information has so far been limited to off-line measurements using labor-intensive methods. We have developed a new technique that has made it possible to monitor VFA on-line in one of the most difficult media: animal slurry or manure. A novel in situ filtration technique has made it possible to perform microfiltration inside a reactor system. This filter enables sampling from closed reactor systems without large scale pumping and filters. Furthermore, due to its small size it can be placed in lab-scale reactors without disturbing the process. Using this filtration technique together with commercially available membrane filters we have constructed a VFA sensor system that can perform automatic analysis of animal slurry at a frequency as high as every 15 minutes. Reproducibility and recovery factors of the entire system have been determined. The VFA sensor has been tested for a period of more than 60 days with more than 1,000 samples on both a full scale biogas plant and lab-scale reactors. The measuring range covers specific measurements of acetate, propionate, iso-/n-butyrate and iso-/n-valerate ranging from 0.1 to 50 mM (6-3,000 mg). The measuring range could readily be expanded to more components and both lower and higher concentrations if desired. In addition to the new VFA sensor system, test results from development and testing of the in situ filtration technique are being presented is this article. PMID- 12569625 TI - Outdoor helical tubular photobioreactors for microalgal production: modeling of fluid-dynamics and mass transfer and assessment of biomass productivity. AB - The production of the microalga Phaeodactylum tricornutum in an outdoor helical reactor was analyzed. First, fluid dynamics, mass-transfer capability, and mixing of the reactor was evaluated at different superficial gas velocities. Performance of the reactor was controlled by power input per culture volume. A maximum liquid velocity of 0.32 m s(-1) and mass transfer coefficient of 0.006 s(-1) were measured at 3200 W m(-3). A model of the influence of superficial gas velocity on the following reactor parameters was proposed: gas hold-up, induced liquid velocity, and mass transfer coefficient, with the accuracy of the model being demonstrated. Second, the influence of superficial gas velocity on the yield of the culture was evaluated in discontinuous and continuous cultures. Mean daily values of culture parameters, including dissolved oxygen, biomass concentration, chlorophyll fluorescence (F(v)/F(m) ratio), growth rate, biomass productivity, and photosynthetic efficiency, were determined. Different growth curves were measured when the superficial gas velocity was modified-the higher the superficial gas velocity, the higher the yield of the system. In continuous mode, biomass productivity increased by 35%, from 1.02 to 1.38 g L(-1) d(-1), when the superficial gas velocity increased from 0.27 to 0.41 m s(-1). Maximal growth rates of 0.068 h(-1), biomass productivities up to 1.4 g L(-1) d(-1), and photosynthetic efficiency of up to 15% were obtained at the higher superficial gas velocity of 0.41 m s(-1). The fluorescence parameter, F(v)/F(m), which reflects the maximal efficiency of PSII photochemistry, showed that the cultures were stressed at average irradiances within the culture higher than 280 microE m( 2) s(-1) at every superficial gas velocity. For nonstressed cultures, the yield of the system was a function of average irradiance inside the culture, with the superficial gas velocity determining this relationship. When superficial gas velocity was increased, higher growth rates, biomass productivities, and photosynthetic efficiencies were obtained for similar average irradiance values. The higher the superficial gas velocity, the higher the liquid velocity, with this increase enhancing the movement of the cells inside the culture. In this way the efficiency of the cells increased and higher biomass concentrations and productivities were reached for the same solar irradiance. PMID- 12569626 TI - Quantification of a novel h-shaped ultrasonic resonator for separation of biomaterials under terrestrial gravity and microgravity conditions. AB - A novel, h-shaped ultrasonic resonator was used to separate biological particulates. The effectiveness of the resonator was demonstrated using suspensions of the cyanobacterium, Spirulina platensis. The key advantages of this approach were improved acoustic field homogeneity, flow characteristics, and overall separation efficiency (sigma = 1 - ratio of concentration in cleared phase to input), monitored using a turbidity sensor. The novel separation concept was also effective under microgravity conditions; gravitational forces influenced overall efficiency. Separation of Spirulina at cleared flow rates of 14 to 58 L/day, as assessed by remote video recording, was evaluated under both microgravity (300 g/L) and product purity (91%). It also resulted in higher reactor productivity (7.37 g/L. h) and butyric acid yield (0.45 g/g). Without on-line extraction to remove the acid products, at the optimal pH of 6.0, the final butyric acid concentration was only approximately 43.4 g/L, butyric acid yield was 0.423 g/g, and reactor productivity was 6.77 g/L. h. These values were much lower at pH 5.5: 20.4 g/L, 0.38 g/g, and 5.11 g/L. h, respectively. The improved performance for extractive fermentation can be attributed to the reduced product inhibition by selective removal of butyric acid from the fermentation broth. The solvent was found to be toxic to free cells in suspension, but not harmful to cells immobilized in the fibrous bed. The process was stable and provided consistent long-term performance for the entire 2-week period of study. PMID- 12569629 TI - Enhancement of 2-phenylethanol productivity by Saccharomyces cerevisiae in two phase fed-batch fermentations using solvent immobilization. AB - The bioconversion of L-phenylalanine to 2-phenylethanol by Saccharomyces cerevisiae in fed-batch experiments has shown that concentrations of 2 phenylethanol of >2.9 g/L have a negative impact on the oxidative capacity of the yeast. Without tight control on ethanol production, and hence on the feed rate, ethanol rapidly accumulates in the culture media, resulting in complete inhibition of cell growth before the maximal 2-phenylethanol concentration of 3.8 g/L, obtained in the absence of ethanol production, could be achieved. This effect was attributed to a cumulative effect of ethanol and 2-phenylethanol, which reduced the tolerance of the cells for these two products. To enhance the productivity of the bioconversion, a novel in situ product recovery strategy, based on the entrapment of an organic solvent (dibutylsebacate) into a polymeric matrix of polyethylene to form a highly absorbent and chemically and mechanically stable composite resin, was developed. Immobilization of the organic solvent successfully prevented phase toxicity of the solvent and allowed for an efficient removal of 2-phenylethanol from the bioreactor without the need for prior cell separation. The use of the composite resin increased the volumetric productivity of 2-phenylethanol by a factor 2 and significantly facilitated downstream processing, because no stable emulsion was formed. The 2-phenylethanol could be backextracted from the composite resin, yielding a concentrated and almost cell free solution. In comparison to two-phase extractive fermentations with cells immobilized in alginate-reinforced chitosan beads, the use of a composite resin was extremely inexpensive and simple. In addition, the composite resin was found to be insensitive to abrasion and chemically stable, such that sterilization with 2 M NaOH or heat was possible. Finally, the composite resin could be produced on a large scale using commercially available equipment. PMID- 12569630 TI - Pulsed addition of limiting-carbon during Aspergillus oryzae fermentation leads to improved productivity of a recombinant enzyme. AB - Fungal morphology in many filamentous fungal fermentations leads to high broth viscosity which limits oxygen mass transfer, and often results in reduced productivity. The objective in this study was to determine if a simple, fed batch, process strategy-pulsed addition of limiting-carbon source-could be used to reduce fungal broth viscosity, and increase productivity of an industrially relevant recombinant enzyme (glucoamylase). As a control, three Aspergillus oryzae fed-batch fermentations were carried out with continuous addition of limiting-carbon. To determine the effect of pulse-feeding, three additional fermentations were carried out with limiting-carbon added in 90-second pulses, during repeated five-minute cycles. In both cases, overall carbon feed-rate was used to control dissolved oxygen concentration, such that increased oxygen availability led to increased addition of limiting-carbon. Pulse-fed fermentations were found to have smaller fungal mycelia, lower broth viscosity, and improved oxygen mass transfer. As a result, more carbon was added to pulse fed fermentations that led to increased enzyme productivity by as much as 75%. This finding has significant implications for the bioprocessing industry, as a simple process modification which is likely to cost very little to implement in most production facilities, has the potential to substantially increase productivity. PMID- 12569632 TI - [Effects of blocking the TGF alpha-EGFR autocrine loop on the growth of human pancreatic carcinoma cells in vitro]. AB - OBJECTIVE: Investigate the effects of blocking the TGF alpha-EGFR autocrine loop on the growth of human pancreatic carcinoma. METHODS: The pCMV-AS-EGFR, a recombinant vector expressing antisense EGFR under the control of human CMV promotor was constructed and transfected the transformant PC-7/AS-TGF alpha cell line cells, which had been transfected by a recombinant retroviral vector expressing antisense TGF alpha. G418 resistant colonies were isolated and identified as PC-7/AS-TGF alpha/AS-EGFR. The integration and expression of exogenous and endogenous genes were detected by Southern blot and Northern blot analysis. Apoptosis was detected by DNA fragmentation, flow cytometry and in situ cell death detection. RESULTS: The cells which were double-transfected by the recombinant vectors showed the integration and expression of exogenous genes, and the downregulation of endogenous EGFR and cyclin D1 mRNA. 125I-EGF ligand binding test showed the binding affinity of the EGFR on the cell surface also reduced. The inhibition effect of cotransfecting of antisense TGF alpha and antisense EGFR was remarkable as compared with that of antisense TGF alpha alone. The incorporation rate of 3H-TdR reduced from 25% to 14.5%, and the growth inhibition rate increased from 78.6% to 86.0%. The ability of soft agar colony-formation was completely suppressed. CONCLUSIONS: These observations strongly support that the blockage of the expression of autocrine growth factor TGF alpha and its receptor EGFR was a potent way in circumventing the malignant properties of the pancreatic carcinoma cells. PMID- 12569631 TI - Inactivation of bacteria and spores by pulse electric field and high pressure CO2 at low temperature. AB - The common methods for inactivation of bacteria involve heating or exposure to toxic chemicals. These methods are not suitable for heat-sensitive materials, food, and pharmaceutical products. Recently, a complete inactivation of many microorganisms was achieved with high-pressure carbon dioxide at ambient temperature and in the absence of organic solvent and irradiation. The inactivation of spores with CO(2) required long residence time and high temperatures, such as 60 degrees C. In this study the synergistic effect of pulsed electric field (PEF) in combination with high-pressure CO(2) for inactivation was investigated. The bacteria Escherichia coli, Staphylococcus aureus, and Bacillus cereus were suspended in glycerol solution and treated in the first step with PEF (up to 25 KV/cm) and then with high-pressure CO(2) not higher than 40 degrees C and 200 bar. The inactivation efficiency was determined by counting the colony formation units of control and sample. Samples of the cells subjected to PEF treatment alone and in combination with CO(2) treatment were examined by scanning electron microscopy to determine the effect of the processes on the cell wall. Experimental results indicate that the viability decreased with increasing electrical field strength and number of pulses. A further batch treatment with supercritical CO(2) lead to complete inactivation of bacterial species and decreased the count of the spores by at least three orders of magnitude, the inactivation being enhanced by an increase of contact time between CO(2) and the sample. A synergistic effect between the pulsed electric field and the high-pressure CO(2) was evident in all the species treated. The new low temperature process is an alternative for pasteurization of thermally labile compounds such as protein and plasma and minimizes denaturation of important nutrient compounds in the liquid media. PMID- 12569633 TI - [Clinical study on antibodies against EBV in sera of patients with rheumatoid arthritis]. AB - OBJECTIVE: Study on the relationship between the infection of Epstein-Barr virus (EBV) and the pathogenesis of rheumatoid arthritis (RA). METHODS: The IgA and IgG antibodies against EBV capsid antigen (IgA/VCA and IgG/VCA respectively), and anti-Z protein IgG antibodies (IgG/Z) in the sera from the patients with RA, systemic lupus erythematosus (SLE), and normal controls were detected by using indirect immunofluoresence and immunobloting techniques. RESULTS: The positive rate of IgA/VCA antibody in the serum of RA patients which was significantly higher than that in SLE patients and normal subjects. The anti IgG/Z antibodies were only found in RA patients with IgA/VCA antibody. CONCLUSIONS: The results suggested that activated EB virus may play a role in the pathogenesis of RA. It is a useful laboratory method for the clinical diagnosis of RA. PMID- 12569634 TI - [The modulation of collagen synthesis and expressions of type I and III collagen mRNAs in aorta smooth muscle cell by PDGF and heparin]. AB - OBJECTIVE: The effect of platelet derived growth factor (PDGF) and heparin on the modulation of human aorta smooth muscle cell(hASMC) proliferation, collagen synthesis, expressions of type I and III collagen mRNAs as well as transforming growth factor-beta (TGF-beta) mRNA were investigated. METHODS: 3H-TdR and 3H proline incorporation and Northern blot analysis were done. 3H-TdR and 3H-proline incorporation was statistically analyzed with t test among different experimental groups. RESULTS: In comparison to the control group, PDGF possessed the ability in promoting markedly the DNA synthesis (3H-TdR incorporation: PDGF group vs control, P < 0.01), synthesis and secretion of collagen protein (3H-proline incorporation of SMC and medium: PDGF group vs control, P < 0.01), expressions of type I and type III collagen mRNAs, and transforming growth factor mRNA in hASMC. Whereas heparin significantly decreased the DNA synthesis (heparin group vs control, P < 0.01), synthesis and secretion of collagen protein (heparin group vs control, P < 0.01) of hASMC in vitro. Heparin also inhibited the promoting effect of PDGF on DNA synthesis, the synthesis and secretion of collagen protein, and reduced the up-regulation of expressions of type I and type III collagen mRNAs as well as TGF-beta mRNA that stimulated by PDGF. CONCLUSIONS: PDGF could promote the collagen synthesis through upregulating type I and type III collagen mRNA expressions while heparin just inhibiting SMC DNA and collagen synthesis effected in the way of anti-atherosclerosis. PMID- 12569635 TI - [Expression of nm23-H1 gene in oral squamous cell carcinoma and its relation to tumor metastasis]. AB - OBJECTIVE: To make an inquiry into the relation between the expression level of nm23-H1 protein and metastasis of oral squamous cell carcinoma (OSCC). METHODS: We evaluated the expression of nm23-H1 protein in 75 cases of OSCC by immunohistochemical technique (SABC). RESULTS: These immunoreactivities of nm23 H1 protein were most common in the cytoplasm of tumor and epithelia cell. Using single-factor chi-test, we found there was a significant correlation between the expression of nm23-H1 protein and lymph node metastasis of OSCC (P < 0.01), pathology grade (P < 0.05). But no statistical association was found among nm23 H1 protein expression level and tumor size, site and patient's sex, age. Using multi-factor logistic stepwise regression analysis, we found that nm23-H1 protein expression level had more effect on OSCC cervical lymph node metastasis than any other parameters. CONCLUSIONS: The results suggested that nm23-H1 protein play an important role in the suppression of the metastasis of OSCC, and that determining the expression level of nm23-H1 protein may help us prognose the metastasis of OSCC. PMID- 12569636 TI - [Preparation of the high-titer retroviral viruses for transducting PDGF-A gene]. AB - OBJECTIVE: To prepare high-titer recombinant retrovirus for transducting platelet derived growth factor A chain(PDGF-A) gene, which will pave the way for future research in vitro and in vivo on biological function of autocrine and paracrine of PDGF-A gene expression product. METHODS: The cDNA fragment of human PDGF-A was inserted into the polyclonal sites of retroviral vector GINa containing cytomegalovirus (CMV) promotor. The constructed recombinant plasmid was transformed into PA317 packaging cell line. The positive clones were obtained by selection in G418-medium and amplified. The supernatant virus was used to infect NIH3T3 cells and titrated. After drug resistance selection, NIH3T3 cell clones were amplified, and a histochemical analysis was performed. SDS-PAGE electrophoresis and Western blot as well as 3H-TdR incorporation methods were adopted to assay the presence and the mitogenic activity of the PDGF in the conditioned medium. RESULTS: The highest titer of the virus was 1.4 x 10(5) CFU/ml. The PDGF-A expression was verified with SDS-PAGE electrophoresis and Western blot and immunofluorescent staining. The cell mitogenic activitiy in the media was up to 51.2 U/(10(6).d). CONCLUSIONS: We had made high-titer recombinant retrovirus for transducting human PDGF-A gene. The gene was expressed with high efficiency. The expressed product has significant mitogenic effect on NIH3T3 cell line. PMID- 12569637 TI - [Modification of N-terminal cDNA of hGM-CSF and high-expression in E. coli]. AB - OBJECTIVE: N-terminal cDNA of human granulococyte-macrophage colony stimulating factor(GM-CSF) was designed to be modified and highly-expressed in E. coli. METHODS: A pair of oligo-nucleotide primers were used to modify the mature N terminal cDNA sequence of hGM-CSF with the method of PCR. the modified cDNA of hGM-CSF was cloned into E. coli. expressive vector PBV220 and expressed in E. coli DH5 alpha strain, the biological activities of the recombinant protein was identified by means of cell colony formation and TF-1 cell growth assay in vitro. RESULTS: The expression level of modified hGM-CSF cDNA was higher than that of unmodified native type. SDS-PAGE revealed that expressed protein of hGM-CSF accounted for about 25% of total bacterial cell protein. The biological activity of recombinant protein was about 1.5 x 10(7) U/mg protein. The sequence of 1-16 amino acid of N-terminal of the recombinant protein was same with native hGM-CSF. CONCLUSIONS: Modification of the N-terminal cDNA of hGM-CSF could dramatically enhance the expression level in E. coli system. PMID- 12569638 TI - [Nucleotide sequence at position -155 to +25 of the 5' flanking region of the angiotensinogen gene of Han people]. AB - OBJECTIVE: To study the polymorphism of 5'-flanking region of angiotensinogen gene and relations to essential hyperfension. METHODS: The nucleotide sequence at position -155 to +25 of the 5'-flanking region in angiotensinogen gene of Han people in Chinese population was identified by applying PCR-single stranded conformational polymorphism(SSCP) and PCR-directed sequencing. RESULTS: (1) The Han people carry an adenylate(A), instead of a cytidylate(C) at position -20 of the 5'-flanking region of AGT gene; (2) A new mutation T-->A at position -46 was detected and A-allele frequencies was similar in both hypertensives and normotensive controls. CONCLUSIONS: The variant T-->A at position -46 of AGT gene was not associated with hypertension, but an adenylate (A) at position -20 of the 5'-flanking region of AGT gene. might be an genetic marker for Hans people. PMID- 12569639 TI - [Dexamethasone inhibits IL-5-induced eosinophil survival]. AB - OBJECTIVE: To assess the effect of dexamethasone (Dex) on interleukine-5-induced survival of eosinophils (EOS) in vitro. METHODS: EOS were purified from guinea pigs by Percoll density gradient and incubated for 4 days in the presence of IL-5 with or without Dex. Cell viabilities were determined by staining cells with fluorescein diacetate and propidium iodide. RESULTS: In the absence of Dex, IL-5 enhanced EOS survival was inhibited in a dosedependent manner. Inhibition was time-dependent and required at least 2 days exposure of EOS to Dex. When EOS were incubated with varying concentrations of IL-5 and 1,000 nmol/l Dex, survival inhibition was reduced at higher concentration of IL-5, and completely abolished by IL-5 1,000 pg/ml. CONCLUSIONS: Dex acts directly on mature EOS to decrease their viabilities, implying the Dex may be important in the treatment of allergic and other inflammatory eosinophilic disorders. PMID- 12569640 TI - [Evaluation of the risk of vertebral fracture predictable with quantitative tibial ultrasound velocity test in postmenopausal women]. AB - OBJECTIVE: To investigate predicting value of increased risk of vertebral fracture associated with reduction of tibial ultrasound velocity (SOS) test in postmenopausal women. METHODS: 193 menopausal women aged(54.4 +/- 7.3) years were included in our study. Post menopausal period of these subjects were (16.0 +/- 7.8) years. These participants received tibial ultrasound speed (SOS) test, DEXA for lumbar 2-4, femoral neck wards triangle and trochant, as well as SPA for 1/3 and 1/10 of distal forearm. Vertebral fracture was determined by radiograph of lateral lumbar and thoracic spine. RESULTS: 54 subjects were found at least one vertebral deformation, and no vertebral deformation was found in other 139 subjects. Tibial SOS was significantly lower in subjects with than without vertebral fracture [(3,651.9 +/- 188.3) vs (3,731.3 +/- 140.8), P < 0.01], The similar results were also found in BMD. The analysis of logistic regression showed: odds ratio (OR) of SOS to vertebral fracture was 1.86(P < 0.01) when 1 standard deviation of SOS changed. It was lower than OR of BMD by DEXA(2.25-3.28, all P < 0.001), and similar with OR of BMD by SPA (1.50-1.96). CONCLUSIONS: Low SOS was associated with presence of vertebral fracture in postmenopausal women. SOS could predict the risk of vertebral fracture, 86% of the risk of vertebral fracture increased when 1 standard deviation of SOS reduced. PMID- 12569641 TI - [An experimental study of viscoelastic properties of articular-cartilage of patella]. AB - OBJECTIVE: To study the viscoelastic properties of human articular cartilages of patella in 8 cadavers, which were obtained from patients ranged in age from nineteen to fifty-one years old and died of acute head injury. METHODS: The creep and stress relaxation of articular cartilage of patella were abserved in the condition of articular cartilage loaded of 500 Pa, just as the physiological load of articular cartilage of patella in keeping human body standing position with the knee joint flexed 30 degrees, during the experiment. The deformation-time curve under certain load, the load-time curve under certain deformity and related data were obtained. RESULTS: Creep and stress relaxation showed that obvious changes occured in the first 10-15 minutes during the whole process. CONCLUSIONS: The data and curve will not only be helpful to understand the pathogenesis of chondromalacia, but also propose a feasible method to study it from biomechanical view, meanwhile provide reference data for designing the prosthesis of patella and material choosing. PMID- 12569642 TI - [The morphometric analysis of lung on protective effect of Chinese herbs 814 on elastase-induced emphysema in hamsters]. AB - OBJECTIVE: To determine the protective effect of Chinese herbal medicine 814 on elastase-induced emphysema in hamsters. METHODS: Animals were injected intratrachealy with elastase for emphysematous models, prophylactic-therapeutic groups were administrate with 814 through esophagus two weeks before the instillation of elastase untill animals were killed at three different time at first, second, and third month. Pulmonary artery pressure, blood gas analysis, heart index and the ratio of dried weight to wet weight of lungs (WW/DW) were examined. The lung paraffin sections were measured mean linear intercept (MLI), mean alveolar number (MAN), ratio of parenchyma area to total area (PA/TA) by the microscope-computer morphometric analysis system. RESULTS: WW/DW in the prophylactic-therapeutic groups was recovered at the same level with the controls, whereas the emphysematous were significantly increased (P < 0.05); and compared with the emphysematous groups, the prophylactictherapeutic groups significantly decreased in MLI, increased in MAN and PA/TA (P < 0.001 or P < 0.05). CONCLUSIONS: Administration of 814 could partly inhibit the development of emphysema induced by elestase in hamsters. PMID- 12569643 TI - [Gene cloning and expression of Mycobacterium leprae alpha 2 antigen]. AB - OBJECTIVES: The recombinant alpha 2 antigen of M. leprae was prepared using the molecular biologic tools and the recombinant DNA expression technology. METHODS: Screening of the M. leprae expression library was performed by the plaque hybridization technique. Nucleotide sequences were determined by dideoxy termination method. RESULTS: The gene coding for alpha 2 antigen of M. leprae was cloned and characterized, and the complete nucleotide sequence data has been assigned in the GSDB, DDBJ, EMBL and NCBI nucleotide sequence databank. The over expression system of alpha 2 antigen gene in E. coli was constructed, and the recombinant alpha 2 antigen has been purified by amylose column chromatography at the purity of more than 95%. More than 10 mg of recombinant alpha 2 antigen has been obtained from 200 ml of liquid culture. CONCLUSION: The recombinant alpha 2 antigen of M. leprae could be used as one of the specific antigens for the sero diagnosis of leprosy. PMID- 12569644 TI - [Clinical features and changing trends of bacterial liver abscess: analysis of 48 cases]. AB - OBJECTIVE: To analyse and compare the changing of clinical feature, pathogenic bacteria, and so is diagnosis and treatment of bacterial liver abscess up date. METHODS: Retrospective study of forty-eight patients over 10 years in PUMC hospital. RESULTS: Compared with the patients in 1960s, the complication and mortality of this disease decreased remarkably. The main clinical manifestations and signs became less typical. Cholelithes and tumor of biliary tract instead of biliary ascariasis became chief pathogenic factor (29.2%). Patients with diabetes mellitus complicating bacterial liver abscess tended to increase (8.3%). The case with negative bacterial culture has increased (52.1%). Ultrasonography should be the first choice for diagnosis, the confirmed diagnosis rate is 79.2%. Percutaneous drainage of the abscess and combined using antibiotics are the effective treatment method. Septic shock was the chief complication causing death. CONCLUSIONS: This disease becomes less typical and difficult to early diagnosis attention should be paid to clinical medicine. PMID- 12569645 TI - [Management of thoracic esophageal perforation]. AB - OBJECTIVE: To compare surgical management of esophageal perforation on thoracic portion with nonoperative management. METHODS: Seventeen patients were treated for thoracic esophageal perforation at our department between 1962 and 1996. Among them, seven patients underwent nonoperative management. The remaining 10 had operative procedures (primary repair in 5, esophagostomy in 3, and drainage alone in 2 cases). RESULTS: Postoperative leakage occurred in 2 patients; among the two leakages, 1 required cervical esophagostomy, and 1 became a controlled fistula needed pleural drainage and feeding jejunostomy. One patient had anastomotic narrowing after esophagostomy. Of patients with nonoperative management, five survived (with 28% in-hospital mortality rate), Whereas only one patient, who was treated by surgical procedure, died (with 10% inhospital mortality rate). The mean hospital stay in nonoperative and operative group were (28.3 +/- 12.9) days and (57.6 +/- 52.7) days respectively. CONCLUSIONS: The clinical observation suggested that rapid diagnosis of thoracic esophageal perforations is essential and once the diagnosis of esophageal perforation is established, a appropriate management must be selected promptly. PMID- 12569646 TI - [Acquirement and identification of a novel intron 1 of human Na(+)-K(+) exchanging ATPase alpha 1 subunit gene]. AB - OBJECTIVE: The unknown part genome structure of the alpha 1 subunit of human Na(+)-K(+)-Exchanging ATPase gene which covers the 80-130 aa in its extracellular domain was studied. METHODS: By means of PCR, the targeted sequences was amplified by using two different templates: human Genome DNA and the muscle cDNA library; the PCR products were analyzed by restriction endonuclease digestion, then cloned into plasmid vector to perform the chemiluminescence Sequencing. The homologous search, splicing site detection and the analysis of the sequence were applied. RESULTS: Two different lengths of products, 833 bp(Fg) and 195 bp(Fc) were acquired after PCR amplification with the human genome DNA and muscle cDNA library respectively. There was an insertion of a 638 bp fragment in Fg at its 138-775 bp. And there was not obviously known sequence homologous to the insertion sequence. CONCLUSIONS: A novel sequence containing the complete human Na(+)-K(+)-Exchanging ATPase alpha 1 subunit gene intron 1 was found, the exact splicing donor and receptor site within it were identified. The Accession No of this sequence is L76938. Analysis of this sequence showed that the intron 1 may function in the regulation of the gene expression and also had a potential coding sequence. PMID- 12569647 TI - [Tissue specificity and expression of a human sperm protein's gene BSD-2.4]. AB - OBJECTIVE: To investigate the tissue specificity and expression in spermatogenic stage of BSD-2.4 cDNA which might be related with fertility and its sequence was received by GenBank. METHODS: Dot blot, Northern blot, and in situ hybridization were used. RESULTS: Northern blot analysis of sixteen human tissues showed that BSD-2.4 cDNA had one transcript only with testis. BSD-2.4 mRNA was detected in germ cells of human testis. CONCLUSIONS: BSD-2.4 is a testis-specific protein. The cDNA was expressed throughout the stages of the spermatogenic cycle. PMID- 12569648 TI - [Relationship between complement activation and reactive oxygen species generation in inflammatory response]. AB - OBJECTIVE: To verify the proposed activation feedback mechanism between complement and polymorphonuclear leukocytes(PMN) derived reactive oxygen species(ROS). METHODS: In vitro tests were designed to identify the activation feedback process by alternative activations of complement by inulin or PMNs by 12 O-tetradecanoylphorbol-13-acetate(TPA) in the test medium in consisting serum complement and isolated PMNs, with chemiluminescence technique to determine the production of ROS and complement fixation trial for complement activations. RESULTS: Evidently that activated complement could activate PMNs to release ROS which further activated complement, so the activating feedback cycle was set. CONCLUSIONS: The complement and PMNs-ROS activation feedback cycle mechanism was recognized. It was expected to serve for delineating the regulation of inflammatory response. PMID- 12569649 TI - [Role of N-methyl-D-aspartat receptor-nitric oxide-cyclic guanosin monophosphate pathway in ischemic brain injury]. AB - OBJECTIVE: To observe the role of NMDA receptor-Nitric Oxide(NOS)-cGMP pathway in ischemic brain injury. METHODS: The common cervical arteries of the rats were transient blocked bilaterally, in association with bleeding from their tails and followed by reperfusion. The procedures were repeated once again as above to establish a stable ischemic brain injury model. 3H-MK801 binding, cNOS activity, iNOS activity, and the cGMP content were measured at different time intervals after ischemic injury. RESULTS: The results indicated that the change of 3H-MK801 binding varied among the cerebral cortex and hippocampus. cNOS activity began to rise in all parts of the brain 24 hours after operation and reached its peak in 3 days. The regions where iNOS and cNOS activity and cGMP content all increased significantly include the hippocamus, striatum and cortex, especially in hippocampus. CONCLUSIONS: The tendency and degree of increase of those four indixes were cosistent during ischemic injury, indicating an important role of NMDA receptor-NO-cGMP in ischemic injury of the hippocampus. PMID- 12569650 TI - [IFN alpha- and IFN gamma-induced differentially expressed genes]. AB - OBJECTIVES: It was previously found that IFN alpha could reduce, while IFN gamma could enhance metastatic potential of a mouse mammary adenocarcinoma cell line MA891. The aim of this study was to identify the sequence of cDNA fragments that were differentially expressed by the two types of IFN, respectively. METHODS: Total RNA was extracted from MA891 cells treated with IFN alpha or IFN gamma. Northern blotting was used to confirm the differentially expressed gene fragments. DNA sequencing was then carried out. RESULTS: The Northern blots indicated that among the 10 cDNA fragments obtained by mRNA differential display technique, 4(G15 gamma, C4 alpha, T2 gamma, and T11 gamma) were shown to be unequivocally differentially expressed. Among them, the nucleotide sequence of G15 gamma did not show homology with any of the existing genes, while that of C4 alpha, T2 gamma and T11 gamma was highly homolgous to mouse minor histocompatility complex gene, mouse beta-actin gene and hamster high blood pressure gene, respectively. CONCLUSIONS: Four differentially expressed cDNA fragments by IFN alpha- and IFN gamma-treated MA891 cells have been ascertained. Regardless of sequence homology, none of them is reportedly related to tumor cell metastasis. The results herein reported provide basis for the elucidation of new metastasis-associated gene(s). PMID- 12569651 TI - [Isolating, cloning, and sequencing of HPV16E6E7 gene from a cervical carcinoma biopsy in Shandong Province]. AB - OBJECTIVE: In order to study the structure specificity of HPV16E6E7 gene of a Chinese patient of cervical carcinoma in Shandong province. METHODS: The tissue DNA were abstracted from cervical carcinoma biopsies and the type of HPV was identified by HPV multiple primers PCR. HPV16E6E7 gene was amplified by PCR from the cervical carcinoma tissue DNA with the infection of HPV16 type only, and then cloned the E6E7 gene into pALTERI vector. After sequencing the double strand, the gene was compared with the prototype E6E7 gene of HPV16. RESULTS: A new recombinant plasmid was constructed and named HPV16E6E7-SD. Sequencing results showed one mutation in HPV16E6E7-SD, the 557th neocleotide in the viral neocleotide sequence "T" was changed into "C", and caused the termination cordon TAA of E6 gene to convert into a Gln cordon CAA. CONCLUSIONS: There is a structure difference between HPV16E6E7-SD and the standard strain. PMID- 12569652 TI - [Synthesis and distribution of type IV collagen in mice testes]. AB - OBJECTIVE: The variations of the synthesis and distribution type IV collagen in the testes of young, adult, and old mice were studied. METHODS: Biotin-avidin DCS system indirect immunofluorescence technique and in situ hybridization method were used. RESULTS: Sertoli cell could synthesize type IV collagen. Type IV collagen was present in the basement membrane of seminiferous tubules in mouse testis. Type IV collagen mRNA signal was seen in the Sertoli cells. The content of type IV collagen and type IV collagen mRNA peaked at day 15, and then declined successively to adult value and the aged value. CONCLUSIONS: The results suggest that the extracellular matrix of testis might play an important role in the process of spermatogenesis. PMID- 12569653 TI - [Effects of antisense N-myc gene on nerve growth factor-induced differentiation of neuroblastoma cell line]. AB - OBJECTIVE: The purpose of this study is to investigate the effects of antisense N myc gene transfection on nerve growth factor-induced differentiation of neuroblastoma cell line. METHODS: Recombinant retroviral vector expressing antisense N-myc gene was constructed. Using the Transfectam Reagent, the recombinant vector was transduced into the human neuroblastoma cell line, IMR 32/NGFR, which was previously transfected by nerve growth factor receptor gene. The transformant cell line expressing antisense N-myc was established and studied by single chain RNA probe hybridization, immunocytochemistry, and nerve growth factor treatment to see whether nerve growth factor can induce differentiation in this transformant cell line and inhibition of N-myc expression. The TUNEL technique and electromicroscopy were used to detect apoptosis of the tumor cells. RESULTS: The cell line transduced by antisense N-myc expressed much less amount of N-myc both on mRNA and protein levels. After nerve growth factor treatment, remarkable morphological differentiation appeared in these cells. Apoptosis was also enhanced in these cells. CONCLUSIONS: Antisense N-myc transfection could specifically inhibit the expression of N-myc and promoted the nerve growth factor induced differentiation of neuroblastoma cell line. Transfection of antisense N myc could also enhance the process of apoptosis of the tumor cells. PMID- 12569654 TI - [Vestibular potentials evoked by electrical stimulation]. AB - OBJECTIVE: To identify the evoked potentials (VsEPs) by electrical stimulation were from vestibule. METHODS: The stimulating electrode was set on the round window of guinea pig. Constant current shocks of 0.05 ms (0.25-1.2 mA) were used to evoke VsEPs by means of vertex-pinna skin electrodes. RESULTS: These potentials were short latencies of (0.973 +/- 0.086) ms, (1.618 +/- 0.176) ms and (2.416 +/- 0.274) ms respectively, which reflected true potentials and were not from electrical stimulus artefacts. It was pure vestibular origin. As being masked with a continuous white noise (120 dB SPL) or removed ipsilateral auditory nerve, and even after facial neurectomy, waves were still existing, but disapeared after selective vestibular neurectomy. The twitching responses in the facial region had never been detected during the whole test. These potentials were bioelectric responses in the vestibular sensory pathway, characterized by threshold saturation, adaptability of excitation, dependent on intact vestibular nerve. CONCLUSIONS: We have set up animal model of recording VsEPs evoked by electrical stimulation on the round window of guinea pigs. PMID- 12569655 TI - [Effect of dialysates on ultrastructure of mouse peritoneal mesothelium]. AB - OBJECTIVE: To observe the effect of various peritoneal dialysates on the ultrastructure of the peritoneal mesothelium. METHODS: Continuous ambulatory peritoneal dialysis mice model was made by injecting domestic lactate, acetate, and American Baxter dialysates intraperitoneally. The diaphragmatic peritoneum of the mice was taken at 10th day and 21th day respectively after injection. RESULTS: After 10 day experiment, the mesothelial ultrastructure of Baxter group was normal, but some pathological changes occurred in the peritoneal mesothelium of both domestic lactate and acetate groups. And the injury of the peritoneal mesothelium was progressing with times, e.g. adhesion and exfoliation of the microvilli, shrinkage of the mesothelial cells and even fibrous adhesion. The extent of effects on the mesothelial cells varied in three groups, the least is Baxter group. After 21 day experiments, the diameters of the stomata were (6.96 +/- 2.46) microns and (6.98 +/- 2.16) microns in both domestic lactate and acetate dialysates groups respectively, which were much greater than those in physiological condition (1.47 +/- 0.88) microns (P < 0.01). The distribution density of the lymphatic stomata were significantly increased in acetate and lactate dialysate groups compared with the control group (P < 0.01). But little effects on the lymphatic stomata were showed after using Baxter dialysate. After discontinuing experiment for 10 days, the impairment of the peritoneal mesothelium recovered in Baxter group and improved to varying degrees in both domestic dialysate groups. The lymphatic stomata were still abnormal in both domestic dialysate groups. CONCLUSIONS: Our findings indicate that lactate and acetate dialysates are more like to elicit peritoneal fibrosis and the increase of distribution density and the enlargement of the lymphatic stomata can significantly enhance their absorption effects and increase their reabsorption quantity of dialysate from the peritoneal cavity during peritoneal dialysis. These changes of the lymphatic stomata and their lymphatic reabsorption are thought to be associated with deterioration of peritoneal function, and to cause ultrafiltration failure in the patients on long-term continuous ambulatory peritoneal dialysis (CAPD) therapy. PMID- 12569656 TI - [Expression of c-fos prooncogene in rat brain induced by an olfactory stimulus]. AB - OBJECTIVE: In order to investigate how widespread the activated regions induced by an olfactory stimulus throughout the central nervous system (CNS), c-fos expressions in CNS of the rat observed, which is known as a combined morphologically and functionally probe. METHODS: Rats were stimulated by iso-amyl acetate for about 10 times, then one hour later sacrificed with perfusion of fixatives. After ABC immunocytochemi cal staining with anti-c-fos polyclonal antiserum, the c-Fos protein labelling areas on sections of the brain were observed. RESULTS: The results showed that the labelling areas were estremely widespread throughout the brain and cervical segments of the spinal cord in the rat, expecially much more in the areas above the level of the pons. Nearly all neuronal structures in the limbic system were labeled in addition to structures of the olfactory pathways, suggesting there were neuronal activities in multiple neural circuits, which may be associated with emotional, somatic and visceral responses to the olfactory stimulus. Next, many brain areas which were associated with mechanisms of attention and central state regulation were widely labeled. Unexpected heavy labellings in deep layers of the superior colliculus, inferior colliculus and pontine nuclei were presumably relevant to integrating activity between sensations and motions. CONCLUSIONS: The present study indicated that a specific olfactory stimulus did induce widespread activities of so-called pan brain networks within the central nervous system. PMID- 12569657 TI - [Evaluation of intraoperative ultrasonography for hepatic neoplasm in surgery]. AB - OBJECTIVE: The purpose of this study was to evaluate the impact of intraoperative ultrasound (IOUS) on the management of patients with neoplasms of the liver. METHODS: Forty-nine patients received operations for liver tumors were examined intraoperatively with 5.0 MHz special ultrasound transducers during surgical exploration of the abdomen. Preoperative imaging studies including percutaneous ultrasound (n = 49), magnetic resonance imaging (n = 11), and computed tomography (n = 34) were taken to compare with intraoperative ultrasonography for the evaluation. RESULTS: Sensitivity for detection of hepatic neoplasms showed in intraoperative ultrasound, percutaneous ultrasound, magnetic resonance imaging and computed tomography as 100%(23/23), 74%(17/23), 74%(14/19), and 75%(6/8) respectively. Specificity showed 100%(26/26), 100%(26/26), 93%(14/15), and 67(2/3) respectively. CONCLUSIONS: Intraoperative ultrasound was the most sensitive and specific method for detection and surgery of liver neoplasms, especially the occult neoplasms and small size lesion (< 2 cm). PMID- 12569658 TI - [Clinic observation on the prothetic results of dental supply implant system]. AB - OBJECTIVE: To make a clinical evaluation by the Dental Supply implant system by three years follow-up study. METHODS: 46 patients treated by Dental Supply implant system were chosen for three years follow-up investigations. RESULTS: The ball-bar framework complete denture was the best and the prostheses for single molars were almost a failure, as the success rate was only 14.3%. CONCLUSIONS: Dental Supply system was not appropriate to restore the single tooth and posterial fixed bridge. PMID- 12569659 TI - [Establishment of A + G ligation mediated polymerase chain reaction and its application in in vivo footprinting study]. AB - OBJECTIVE: To establish a new ligation mediated PCR method for the work in in vivo footprinting study of some regions of low G-residue content or some regions of G-residue free. METHODS: The A > G chemical cleavage method of Maxam-Gilbert sequencing was modified for ligation mediated PCR after A + G chemical cleavage and separate the PCR products by sequencing PAGE. RESULTS: A new A + G ligation mediated method was established and could be used in vivo footprinting study. CONCLUSIONS: Our results indicated that the newly developed method could analyze the DNA-Protein interaction at both A-residue and G-residue, enabled us to get more information and enlarged the scope of its application. PMID- 12569660 TI - [The characterization of a novel testis-specific nucleoporin gene]. AB - OBJECTIVE: To investigate the structure and function of testis-specific gene and spermatogenesis in human. METHODS: Screening cDNA expression library, 5' rapid amplification of cDNA ends, Northern blot and fluorescent in situ hybridization(FISH) were used. Gene expressing, purified of expressed protein by affinity chromatography and SDS-PAGE as well as phosphorylation of expressed protein in vitro by PKC and p34cdc2 were observed. RESULTS: A cDNA designated as BS-63 was isolated and found to consist of 2,209 bp with an open reading frame of 2,100 bp and assigned the accession number U64675 by GenBank. The deduced polypeptide consisted of 700 amino acid residues containing XFXFG or FG motifs that were characteristic of nuclear pore complex (NPC) protein and acted as potential binding sites for Ran. The N-terminal region had high homology with Ran BP2/Nup 358, a nucleoporin component, showing that BS-63 was a member of the NPC family. Northern blot analysis of mRNA prepared from various human tissues showed that BS-63 gene was transcribed in two forms: 6.0 and 8.5 kb. The 8.5 kb transcript was present in low amounts in several somatic tissues; whereas the 6.0 kb transcript was expressed only in testis. Analysis by FISH method mapped the BS 63 gene in 2q11.2-12. A protein band with an estimated Mr of 80,000 was detected with E. coli BL21 (DE3) transfected with recombinant plasmid pET30a (+)-BS-63. In vitro phosphorylation test indicated the BS-63 recombinant protein could be phosphorylated by PKC and p34cdc2. CONCLUSIONS: The study was the first demonstration that the BS-63 gene encoding a nucleoporin-related protein with Ran binding sites was expressed in germ cells of human testis. PMID- 12569661 TI - [Differential diagnosis and operative timing of intramedullary spinal cord tumors]. AB - OBJECTIVE: To discuss the differential diagnosis of various kinds of intramedullary spinal cord tumors, and choose the right operative timing. METHODS: On grading the dysfunction of the movement, an analysis has been made statistically for 147 cases with 160 tumors resected by microneurosurgery and confirmed pathologically. Their neuroimage have been also compared. RESULTS: The clinical manifestations of main kinds of intramedullary spinal cord tumors are not significantly different, but there are certain characteristic features in their neuroimage. The tumors of grade 0 group (normal movement) are obviously smaller than those of other grades groups. The pre- and post-operative grades of motor disturbance show a better, positive linear correlation. CONCLUSIONS: MRI, particularly its enhancement, can differentiate an ependymoma from astrocytoma and hemangioblastoma in most cases. A satisfactory result can be achieved to resect the tumor immediately by using the minimally invasive, microsurgical techniques. PMID- 12569663 TI - [Construction of a pufferfish gridded cosmid library]. AB - OBJECTIVE: Pufferfish is a newly-established model organism in human genome research, which has been demonstrated its unique advantages in comparative genomics. Construction of a genomic library of pufferfish is the prerequisite to any studies of its genome. METHODS: Genomic DNA from Fugu rubripes, a species distribute only in Japan and China seas, was used to construct a library in a novel cosmid vector, named sCOGH2, which contains an exon-trapping cassette. RESULTS: This library is composed of 57,600 clones gridded in 60,096-well cell culture plates, one clone each well. The average size of the inserts is around 35 kb. It provides 99.2% probability to find any unique DNA fragment of pufferfish in this library. The clones of this library can survive after more than 10 times freeze-thaw, also demonstrating high stability in propagation. The clones shone highly positive signals when hybridized with the pufferfish genomic DNA. CONCLUSIONS: The library accords with qualitative demands of a cosmid library. PMID- 12569662 TI - [Subtractive cDNA cloning and analysis of murine erythroid terminal differentiation related factor]. AB - OBJECTIVE: Cloning and identification of cDNA related to erythroid terminal differentiation factor (MEDRF). METHODS: cDNA related to erythroid terminal differentiation from the Friend virus anemia (FVA) infected splenic erythroblasts of BALB/c mice were performed by using subtractive hybridization combined with PCR technique. The splenic proerythroblasts isolated were cultured in the presence of erythropoietin for 36 hrs. Subtractive cDNA clones of differentially expressed in the 36 hrs erythroblasts (sub cDNA-36) but absent in the uncultured proerythroblasts were observed. The sub cDNA-36 was then used for construction of subtractive cDNA library using the Bluescript-SK(+) phage vector system, and differentially screened by 32P-labelled PCR generated probes. The positive clones were analyzed by nucleotide sequencing. RESULTS: The results indicated that a 472 bp cDNA fragment which contained a 309 bp reading frame from 51 to 359 coding 102 amino acids was identified and it has been accepted by GenBank as a new cDNA sequence that without comparable homology of existing sequences (accession number: AA114369). Northern blot analysis demonstrated that it was differentially expressed in the 36 hrs cultured intermediate-late stages of erythroblasts. CONCLUSIONS: The newly found cDNA, which expresses specifically in intermediate late stages of erythroblasts, not in stages of proerythroblasts, may be a new gene related to murine erythroid terminal differentiation. PMID- 12569664 TI - [Effects of 17 beta-estradiol on intracellular free calcium, inositol-1,4,5 trisphophate and calmodulin in human osteoblast-like osteosarcoma cell line TE85]. AB - OBJECTIVE: To study the effects of 17 beta-estradiol (E2) on intracellular free calcium ([Ca2+]i), inositol-1,4,5-trisphophate (IP3) and calmodulin (CaM) in human osteoblast-like cell line TE85. METHODS: Using Fluo-3/AM as fluorescent indicator, the [Ca2+]i was measured by laser confocal microscopy system. The IP3 content was determined by anion-exchange chromatography. CaM content was detected by a high sensitive assay based on stimulation of Ca(2+)-dependent phosphodiesterase activity. RESULTS: E2 at dose of 0.1 and 1.0 nmol/L increased fluorescent level by 4.7 and 6.1 times. Pretreatment with thapsigargin (100 nmol/L), the E2 caused only 1.5 times elevation in fluorescence. E2 induced a concommitant bi-peak increase in IP3 content. At the presence of E2(1.0 nmol/L), the CaM content increased by 85.2%. Tamoxifen did not affect the effect of E2 on [Ca2+]i, IP3 and CaM content. But, the inhibitor of phospholipase C (neomycin) and pertusis toxin depressed them partly or completely. CONCLUSIONS: E2 regulate bone cells function by way of Ca2+/CaM activation. PMID- 12569666 TI - [C677T genetic polymorphism of methylenetetrahydrofolate reductase in premature coronary heart disease]. AB - OBJECTIVE: Methylenetetrahydrofolate reductase (MTHFR) is an important factor responsible for hyperhomocysteinemia. The relation of MTHFR gene C677T polymorphism and premature coronary heart disease was studied. METHODS: MTHFR C677T genetic polymorphisms in 67 patients with premature coronary heart disease were detected by PCR-RFLP technique. RESULTS: In case group, the frequency of T homogenic type was 34.3% (23/67), heterogenic type 43.3% (29/67) and C homogenic type 22.4% (15/67). T allele frequency was 55.9% (75/134) while C allele frequency 44.1% (59/134) in case group. There were significant differences in MTHFR genotype and allele frequencies between cases and controls (chi 2 = 6.82 and 5.41 respectively, P < 0.05). CONCLUSIONS: It was suggested that MTHFR gene C677T mutation was a possible risk factor of Chinese premature coronary heart disease. PMID- 12569665 TI - [Experimental studies in vitro on the alpha-fetoprotein-specific promoter mediated target gene therapy of hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the alpha-fetoprotein (AFP)-specific promoter mediated target gene therapy of hepatocellular carcinoma in vitro. METHODS: Retroviral vector (LX2.2CD) in which the cytosine deaminase (CD) gene was driven by the 2.2 kb recombinant human AFP TRS. After transfecting three human hepatoma cell lines and one non-hepatoma cell line with LX2.2 CD, anti-G418 clones integrated CD gene were selected, and inhibitory experiment of cell growth was performed. RESULTS: 5 fluorocytosine (5FC) could confer the chemosensitivity to transduced AFP producing hepatoma cells (HuH-7 and huH-1/c1-2), but not to AFP-nonproducing hepatoma cells (HLE) or nonhepatoma cells (GLC). On the other hand, when transfecting the above four tumor cell lines with another retroviral vector pCD2 (CD gene was driven by 5'LTR internal promoter), no cell selection was found in 5FC-induced cell grow inhibition. CONCLUSIONS: Recombinant retrovival transfer of the CD gene under the control of the AFP TRS followed by 5FC may well be a promising trageted gene therapy for hepatoma. PMID- 12569667 TI - [Apoptosis of human pancreatic carcinoma cell lines induced by combinations of chemotherapeutic drugs]. AB - OBJECTIVE: To investigate the apoptotic in human pancreatic carcinoma cell lines P3, SW1990 and Capan-2 induced by the combined treatment of 5-fluorouracil (5FU), mitomycin C (MMC), carboplatin(CBP) or adriamycine (ADM) or epirubicin (E-ADM). METHODS: The changes of cell morphology were monitored by means of microscopic and fluorescence technique after treated with the combinations of chemotherapeutic drugs. The rate of apoptosis was calculated with flow cytometry and the fragmentation of DNA was measured by agarose gel electrophoresis. RESULTS: The combinations of chemotherapeutic drugs were able to induce the apoptosis of human pancreatic carcinoma cell lines. Compared with the cell necrosis, the dosage of drugs was less and the time was shorter for inducing apoptosis. With the increased dosage and/or the time prolonged for treating the cells, the rate of apoptotic cells was increased. However, the apoptosis reaction of various human pancreatic carcinoma cell lines, which were mentioned above, induced by the same combinations of chemotherapeutic drugs were various different. CONCLUSIONS: The apoptosis process might be a mechanism of an inhibiting cancer cell growth induced by the combinations of chemotherapeutic drugs. The examination of apoptosis reaction for the cells induced by different methods and/or chemotherapeutic drugs could become an important index of testing their ability of inhibiting pancreatic neoplasm. PMID- 12569668 TI - [Distribution of human hsp90 beta gene in active chromatin upon heat shock]. AB - OBJECTIVE: To study the distribution of human hsp90 beta gene in active chromatin of Jurkat cells subjected to heat shock. METHODS: Active and inactive chromatin were isolated by using organomercurial affinity chromatography from Jurkat cells with or without heat shock. Slot-blot hybridization was then carried out with human hsp90 beta cDNA probe. And the efficiency of mRNA expression was studied by Northern blot hybridization. RESULTS: The content of the hsp90 beta gene distributed in the active chromatin was increased upon heat shock which was comparable with the highly efficient mRNA expression induced by the same treatment. CONCLUSIONS: The results suggested chromatin activity was a prerequisite for heat shock induction of hsp90 beta gene. PMID- 12569669 TI - [The role of ICAM-1 in leukocyte adherence to endothelial cells induced by hypoxia/reoxygenation]. AB - OBJECTIVE: To determine whether intercellular adhesion molecule-1 (ICAM-1) mediats the polymorphonuclear leukocyte (PMN) adherence to endothelial cells elicited by hypoxia/reoxygenation (H/R). METHODS: Human umbilical vein endothelial cells (HUVEC) monolayers was exposed to hypoxia for 30 min or 120 min then reoxygenated. Count the number of PMN in adhesion assays. Expression of ICAM 1 and ICAM-1 mRNA was detected by immunohistochemical analysis and in situ hybridization. RESULTS: PMN adhesion to HUVEC exposed to H/R was significantly increased (1 fold) as compared with that of control (P < 0.01). H/R induced hyperadherence was significantly diminished by monoclonal antibodies directed at ICAM-1 or CD11a/CD18. The results showed an increased expression of ICAM-1 and ICAM-1 mRNA on HUVEC exposed to H/R. Hypoxic HUVEC might be responsible for PMN adesion. CONCLUSIONS: ICAM-1 mediated the adhesive interaction between PMN and VEC induced by hypoxia/reoxygenation. PMID- 12569670 TI - [Effect of 15-methyl PGF2 alpha methyl ester combined with tyrosine hydrazide on early pregnancy in mice]. AB - OBJECTIVE: To observe the effect of 15-methyl PGF2 alpha methyl ester (PG05) in combination with tyrosine hydrazide (TH) on early pregnancy in mice. METHODS: The pregnancy rate of mice treated with PG05 and TH was counted and serum progesterone level was measured by RIA. RESULTS: Administration of TH at 90 mg.kg.day-1 to mice on day 7-9 of gestation enhanced significantly the termination of early pregnancy induced by PG05 at 0.2 mg-1.kg-1 for 2 days. Serum progesterone level of mice treated with PG05 combined with TH was found to be markedly lower than that of animals given PG05 or TH alone. TH at 30 micrograms 1.ml-1 inhibited significantly the activity of 3 beta-HSD of luteal cells in rat. However, PG05 at 10 micrograms-1.ml-1 had no influence upon inhibitory effect of 3 beta-HSD activity induced by TH. TH at 150 mg-1.kg-1 for 3 days did not affect the progesterone receptor level of utering cytoplasma in pseudopregnant rats. CONCLUSIONS: The inhibitory effect of TH on the production of progesterone may be the main mechanism to terminate early pregnancy when used synergically with PG05. PMID- 12569671 TI - [Establishment of a microbial assay for screening anti-human immunodeficiency virus type-1 protease inhibitors]. AB - OBJECTIVE: This study was to establish a microbial assay of human immunodeficiency virus type-1 protease (HIV-1 PR) activity for screening anti-HIV PR inhibitors. METHODS: A 24 bp synthetic oligonucleotide fragment that encodes the HIV-1 PR recognition sequence was inserted into the tetr gene of pBR322 (mtetr). Escherichia coli containing HIV-1 PR expression vector-pPOLO was transformed with pACYC184M containing modified mtetr gene. The transformant could express both HIV-1 PR and the modified Tet protein. RESULTS: The growth of engineered E. coli was prevented in the presence of tetracycline because the resistance Tet protein was degraded by HIV-1 PR. However inhibition of the HIV-1 PR restored tetracycline resistance. 31 chemical synthetic compounds were tested by the microbial assay. CONCLUSIONS: A microbial assay method of HIV-1 PR activity was established through a engineered E. coli. 5 mumol/L saqunavir-a special HIV-1 PR inhibitor showed inhibitory effect on the engineered E. coli. That means this model could be used as a initial screening model for anti-HIV PR agents. PMID- 12569672 TI - [The surgical treatment of congenital kyphosis]. AB - OBJECTIVE: To summarize the treatment experience of congenital kyphosis, compare the results of several surgical procedures and conclude the lessons of failure cases with surgical treatment. METHODS: Retrospective analysis of 33 patients with surgical treatment of congenital kyphosis. These patients had a follow-up of 6 months to 14 years with an average follow-up of 5 years and 6 months. RESULTS: Among the 33 cases, the average age at surgery was 16.5 years old and the average kyphosis was 90.2 degrees. Four had anterior fusion, and 7 had posterior fusion only, and 22 had combined anterior and posterior fusion. The percentage of correction of the 3 procedures above mentioned was 32.7%, 34.4% and 41.2% respectively. All of the patients with incomplete paraplegia, but 2 had been recovered completely. CONCLUSIONS: The combined anterior and posterior fusion had a better correction than the anterior group or posterior group alone. PMID- 12569673 TI - [Content variety of alisol B 23-acetate in Rhiozma Alismatis reaped at different time]. AB - Alisol B 23-acetate in Rhizoma Alismatis reaped at different time was determinated by RP-HPLC. The result indicated that the content of those reaped in April is obviously higher than those reaped from Jan. to March. PMID- 12569674 TI - [Identification of crude drugs from genus Leonurus]. AB - The structure in cross section of stem and lamina of 9 species and 1 variety from genus Leonurus, whose herb used as Yimu Cao, were reported. Diagnostic characteristics were listed out in a key for identification. PMID- 12569675 TI - [Pharmacognostical identification on the stem and leaf of Humulus scandens]. AB - In this paper, pharmacognostiacal identification of Humulus scandens was studied. The character of medicinal materials, histological and powder characteristics for the setem and leaf of H. scandens were mainly reported. The microscopical features for leaf is typical identified basis. PMID- 12569676 TI - [Identification of 16 kinds of mineral drugs with fluorescence spectra]. AB - In this paper, 8 groups 16 kinds of mineral drugs were identified by fluorescence spectra. The results showed that the fluorescence spectra method in identifying mineral drugs is not only exact, sensitive, but also using little experimental materials and better repealability. This method can specially be used to identify the mineral drugs which have similar natrues and difficult to be distinguished from thim. PMID- 12569677 TI - [Constituents of volatile oils from Geranium wlassowianum]. AB - In this paper, the chemical constituents of voltile oils from Geranium wlassowianum were analyzed by GC-MS. 24 constituents in the oils were identified (75.42% of total oil), saturated alkanes of them are main components. PMID- 12569678 TI - [Research on the mechanism of Yin-enriching and Qi-reinforcing, strengthening body and tonification deficiency syndrome action of radix Panacis quinquefolii]. AB - Radix Panacis Quinquefolii could significantly promote 3H-TdR infiltrating of spontaneously and ConA-stimulated splenic lymphocytes of mice; Promote the splenic lymphocytes capacity of mice in interleukin2 (IL-2) production; Significantly enhance erythropoitin (EPO) level of mice in serum; Markedly promote production of colony stimulating factors (CSFs) in murine spleen conditioned medium (SCM); And promote production of CSFs in lung-conditioned medium (LCM) of mice. The results suggest that yin-enriching and qi-reinforcing, strengthening body and tonification deficiency syndrome action of Radix Panacis Quinquefolii is in association with its lymphocytes activation-stimulating, IL-2 secrection-promoting, EPO secrection, and CSTs production enhancing effectes. PMID- 12569679 TI - [Effect of radix Achyranthis bidentatae on memory and endurance]. AB - After P. O. decoction of Radix Achyranthis Bidentatae, continuously for seven days in mice, the drug could effectively improve the acquisition of memory of mice, significantly enhance the endurance of mice. The results indicated that the decoction of Radix Achyranthis Bidentatae has the actions of enhancing memory and endurance. PMID- 12569680 TI - [Gastroenteric absorptive kinetic study on strychine in Jiufensan]. AB - After P. O. Jiufensan, the rest curve of strychine in stomach and intestines of mice per kg was determined by TLC scanning. And the gastroenteric absorptive kinetic rule of strychine in rivo was studied, As a result, the strychine in stomach and intestine of mice accorded with pharmacokinetic model of one room. Also the main pharmacokinetic parameters of this preparation were examined. PMID- 12569681 TI - [An experimental study on protective effects of Schwann cell cooperation with ECM gel to neurons in rat dorsal root ganglion]. AB - OBJECTIVE: To research the protective effect of Schwann cell and extracellular matrix (ECM) gel on neurons in dorsal root ganglion. METHODS: 1. Schwann cells were seeded into 30% ECM at 1 x 10(8)/ml and then implanted into PLA hollow fiber conduits to repair 10 mm length defects of rat sciatic nerve, and histological observation was taken at 8 and 12 weeks after operation. 2. To observe the survival of Schwann cells, Schwann cells labeled BrdU were seeded into 30% ECM at 1 x 10(8)/ml and then implanted into PLA hollow fiber conduits to repair 10 mm length defects of rat sciatic nerve. Histological observation and immunohistochemical method stained with BrdU were done at 3 and 6 weeks after operation. RESULTS: 1. When seeded into ECM gel and transplanted into rats, most of the Schwann cells survived to 3 weeks and a part of them survived up to 6 weeks. 2. The survival neuron ratios of Schwann cells with ECM gel group and ECM gel group were 83.5% and 81.3% respectively, and significantly higher than that of saline group (72.9%, P < 0.05). CONCLUSION: When seeded into ECM gel and transplanted into rats, most of the Schwann cells survive and protect 83.5% neurons in dorsal root ganglion from retrograde death. PMID- 12569682 TI - [Study on sense neurotrophism of denervated red and white muscles]. AB - OBJECTIVE: To clarify the character of nerve growth factor (NGF) in the denervated red and white muscles and the relationship between the amount of NGF and sensitive neurotrophism of denervated red and white muscles. METHODS: The model of the denervated gastrocnemius and soleus was made by clipping the sciatic nerve of Wister rats. The immunohistochemistry was taken to measure the amount of NGF in muscles, and the neurotrophism of extracts of muscles was tested with culture of dorsal root ganglions at the 1st, 3rd, 7th and 14th days after injury. RESULTS: The amount of NGF in denervated gastrocnemius and soleus decreased, especially in soleus. The neruotrophism of the extracts of the two kinds of denervated muscles did not decrease; on the contrary, it increased after a week after injury. CONCLUSION: The injury of peripheral nerves causes the amount of NGF in the target tissues to increase, but the change is different between the denervated muscles; the neurotrophism of the extracts of musclesis determined by all kinds of neurotrophic factors, and can not be explained by a single factor. PMID- 12569683 TI - [Effect of interleukin-1 on expression of interleukin-1 receptor in KM mice sciatic nerves before and after injuries]. AB - OBJECTIVE: To study the expression of interleukin-1 receptor in normal mice sciatic nerves and in transected sciatic nerves treated with or without extrinsic interleukin-1 locally at different periods. METHODS: Seventy-two KM mice were equally divided into two groups. All the left sciatic nerves were transected. The stumps in experimental group were soaked in liquid with interleukin-1, whereas those in control group without interleukin-1. Then all the stumps were repaired end to end. At the 3rd hour, 1st day, 3rd day, 7th day, 14th day, and 28th day after operation respectively, every proximal stump was dissected and the expression of interleukin-1 receptor was carried out by immunohistochemistry method (LSAB method). The expression level of interleukin-1 of ten normal sciatic nerves of mice was studied, too. RESULTS: Normal nerves showed interleukin-1 receptor expression on the membrane of Schwann cells. After nerve injury, the interleukin-1 receptor expression increased biphasically in both groups, but the intensity of increase was lower in the experimental group than in the control group. CONCLUSION: Schwann cell is the target cell of interleukin-1. PMID- 12569684 TI - [Application of alpha-cyanoacrylate medical adhesive in fixation of intratemporal facial nerve within chitin chamber]. AB - OBJECTIVE: To study the feasibility of alpha-cyanoacrylate medical adhesive in fixation of intratemporal facial nerve when nerve was repaired within chitin chamber, and to investigate the nerve regeneration. METHODS: Nerve defect of 6 mm was made in left intratemporal facial nerves of 48 rabbits. All the defects were bridged with chitin chamber and were fixed by alpha-cyanoacrylate medical adhesive, surgical suture and natural union. Nerve function test and histomorphological examination were carried out at 1 month and 3 months after repair. RESULTS: It was observed that the nerve was fixed firmly to the chamber with no crack or crease by alpha-cyanoacrylate medical adhesive. The regenerated new nerve fibers were more regular and denser and the neurological function recovered much better in the group fixed by alpha-cyanoacrylate medical adhesive than in the groups those fixed by surgical suture and natural union. CONCLUSION: The medical adhesive is strong in adhesion and beneficial to nerve repair; repair of intratemporal facial nerve defect within chitin chamber fixed by alpha cyanoacrylate medical adhesive is feasible, simple and timesaving. PMID- 12569685 TI - [Repair of peripheral nerve defect by xenogeneic acellular nerve basal lamina scaffolds]. AB - OBJECTIVE: To explore the possibility to bridge peripheral nerve defects by xenogeneic acellular nerve basal lamina scaffolds. METHODS: Thirty SD rats were randomly divided into 5 groups; in each group, the left sciatic nerves were bridged respectively by predegenerated or fresh xenogeneic acellular nerve basal lamina scaffolds, autogenous nerve grafting, fresh xenogeneic nerve grafting or without bridging. Two kinds of acellular nerve basal lamina scaffolds, extracted by 3% Triton X-100 and 4% deoxycholate sodium from either fresh rabbit tibial nerves or predegenerated ones for 2 weeks, were transplanted to bridge 15 mm rat sciatic nerve gaps. Six months after the grafting, the recovery of function was evaluated by gait analysis, pinch test, morphological and morphometric analysis. RESULTS: The sciatic nerve function indexes (SFI) were -30.7% +/- 6.8% in rats treated with xenogeneic acellular nerve, -36.2% +/- 9.7% with xenogeneic predegenerated acellular nerve, and -33.9% +/- 11.3% with autograft respectively (P > 0.05). The number of regenerative myelinated axons, diameter of myelinated fibers and thickness of myelin sheath in acellular xenograft were satisfactory when compared with that in autograft. Regenerated microfascicles distributed in the center of degenerated and acellular nerve group. The regenerated nerve fibers had normal morphological and structural characters under transmission electron microscope. The number and diameter of myelinated fibers in degenerated accellular nerve group was similar to that of autograft group (P > 0.05). Whereas the thickness of myelin sheath in degenerated accellular nerve group was significantly less than that of autograft group (P < 0.05). CONCLUSION: The above results indicate that xenogeneic acellular nerve basal lamina scaffolds extracted by chemical procedure can be successfully used to repair nerve defects without any immunosuppressants. PMID- 12569686 TI - [Treatment of common fibular nerve secondary compression syndrome]. AB - OBJECTIVE: To investigate the mechanism, diagnosis, and treatment of common fibular nerve compression syndrome secondary to sciatic nerve injury. METHODS: Based on the clinical manifestation and Tinel's sign at fibular tunnel, 5 cases of common fibular nerve secondary compression following sciatic nerve injury were identified and treated by decompression and release of fibular tunnel. All 5 cases were followed up for 13-37 months, 25 months in average, and were evaluated in dorsal flexion strength of ankle. RESULTS: The dorsal flexion strength of ankle in 4 cases increased from 0-I degrees to III-V degrees, and did not recover in 1 case. CONCLUSION: Fibular tunnel is commonly liable to fibular nerve compression after sciatic nerve injury. Once the diagnosis is established, either immediate decompression and release of the entrapped nerve should be done or simultaneous release of fibular tunnel is recommended when the sciatic nerve is repaired. PMID- 12569687 TI - [Role of thyroid hormone in peripheral nerve regeneration]. AB - OBJECTIVE: To review the role of thyroid hormone in the peripheral nerve regeneration. METHODS: The recent literatures of experimental study and clinical application on the role of thyroid hormone in nerve regeneration were reviewed. The researches on expression, isoform and changes of thyroid hormones in rat sciatic nerve in normal or injury were summarized. The effect of thyroid hormone on local rat sciatic nerve was studied, too. RESULTS: Nuclear thyroid hormone receptors expressed in numerous nuclei of sciatic nerve during a limited period of development extending from the third week of embryonic life to the end of the second postnatal week and after injury of adult sciatic nerve. A single and local administration of thyroid hormone at the level of the transected sciatic nerve produced a lasting effect on peripheral nerve regeneration. CONCLUSION: The beneficial effects of thyroid hormones upon injured peripheral nerve may have considerable therapeutic potential. PMID- 12569688 TI - [Experimental study on forms of microcirculation of arterialized venous flap in rabbits]. AB - OBJECTIVE: To study the forms of microcirculation of arterialized venous flap. METHODS: Twenty New Zealand rabbits were equally divided into two groups, arterialized venous flap group (group A) and control group (group B). The microcirculatory haemodynamic of arterialized venous flap was studied through observation of transparent chamber in rabbit's ears with aspecial TV set with manification of 1000. RESULTS: The blood of arterilized venous flap flowed through venule anastomosis and drained to another venule. CONCLUSION: It is the main form of microcirculation in early stage that blood flows from venule to draining venule by way of communicating networks between venules. PMID- 12569689 TI - [Acceleration of burn wound healing with topical application of recombinant human epidermal growth factor ointments]. AB - OBJECTIVE: To investigate the efficiency of recombinant human epidermal growth factor (rhEGF) on burn wound healing and to explore the effective density of the ointments. METHODS: A total of 120 cases of burn in superficial II degree and profound II degree were randomly divided into 2 groups. In the first group of 15 cases of superficial II degree, the wounds were treated by rhEGF ointments of different density, 0.5 microgram/g, 10 micrograms/g and 50 micrograms/g, to screen out the effective density. And in the other 105 cases of the second group, optimal density of the ointments based on the result of the first group were employed to treat the burn wound in superficial II degree and profound II degree, with the self-corresponding wounds of the same degree as control, to study the efficiency of rhEGF on wound healing, according to the wound healing time, and adverse reaction of the ointment. RESULTS: In the first group, the average healing time of superficial II wound treated by ointments of 10 micrograms/g and 50 micrograms/g significantly shortened when compared with that treated by ointments of 0.5 microgram/g(P < 0.01), but there was no obvious difference between the cases treated by ointments of 10 micrograms/g and 50 micrograms/g. In the second group, the healing time of superficial II wound treated by ointments of 10 micrograms/g was (8.39 +/- 2.25) days, (9.52 +/- 2.56) days in the control (P < 0.01); and healing time of profound II burn treated by ointments of 10 micrograms/g was (16.80 +/- 2.99) days, (18.27 +/- 3.17) days in the control (P < 0.01). And healing rates of burn wound at different periods were higher than those of the control. CONCLUSION: The above results indicate that rhEGF ointments can enhance burn wound healing significantly, and the ointment of 10 micrograms/g is a good choice for clinical application. PMID- 12569690 TI - [Application of pre-fabricated free flap in reconstruction and repair of skin defect of foot in weight-bearing area]. AB - OBJECTIVE: To investigate the clinical effect of pre-fabricated free skin flap in reconstruction and repair of skin defect of foot in weight-bearing area. METHODS: Eight cases of skin defect of foot in weight-bearing area, due to trauma, were repaired by such an approach; free skin flap was designed and pre-fabricated at the contralateral plantar center, and 3 weeks later the free skin flap, with sensory nerve was transplanted to the site of skin defect, fixed by stitches through drilled holes in the calcaneous bone. All of 8 cases were followed up for 15 to 23 months before clinical evaluation. RESULTS: The wound healed well with no ulcer or deformity. According to American AOFAS scoring standard, it was more than 80 in 5 cases, more than 75 in 2 cases and 70 in one case. CONCLUSION: The pre-fabricated free skin flap from the contralateral plantar center is a good option to reconstruct and repair the skin defect of foot in weight-bearing area, with low ulcer occurrence and good contour. PMID- 12569691 TI - [Repair of soft tissue defect at finger-tips by cross-arm skin flap with lateral ante-branchial cutaneous nerve]. AB - OBJECTIVE: To summarize the application of cross-arm skin flaps with lateral antebranchial cutaneous nerve in repair of soft tissue defect. METHODS: From March 1996 to March 2001, 37 cases of soft tissue defect at fingertips were repaired by cross-arm skin flaps with lateral antebranchial cutaneous nerve, 1.5 cm x 1.5 cm to 3.5 cm x 4.0 cm in size. All of the cases were followed up for 3 48 months with routine evaluation of the wound and the function of hands. RESULTS: All of the flaps survived and the wound achieved primary healing. The sensation and shape of hands recovered well. CONCLUSION: It's a good choice to repair soft tissue defect at fingertips by cross-arm skin flaps with lateral antebranchial cutaneous nerve. PMID- 12569692 TI - [Application of reversed digital artery cross-finger flap with a compound skin pedicle]. AB - OBJECTIVE: To summarize the application of reversed digital artery cross-finger flap with a compound skin pedicle in soft tissue defect of hand. METHODS: From October 1997, 35 fingers of 30 cases, with soft tissue defect at the dorsal side of digital interphalangeal joint and at the fingertips, were repaired by the reversed artery cross-finger flap with a compound skin pedicle, 1.5 cm x 1.0 cm to 2.0 cm x 2.0 cm in size. All of the cases were followed up for 1-6 months and evaluated clinically. RESULTS: All of the flaps survived, with a good texture and no swelling; and there was no adverse side effect on the donor site. CONCLUSION: Reversed digital artery cross-finger flap with a compound skin pedicle is a good option to repair the soft tissue defect of hand. PMID- 12569693 TI - [Management of extensive closed internal degloving injury]. AB - OBJECTIVE: To study the management of extensive closed internal degloving injury (CIDI). METHODS: From September 1987 to October 1999, 18 cases of CIDI were retrospectively reviewed. Of 18 cases, there were 7 cases in thigh, 6 cases in legs and 5 cases in pelvis, ranging from 15 cm x 12 cm to 38 cm x 25 cm in size. Various managements were adopted according to the severity of the injury, including vacuum drainage and adjuvant compression in 5 cases, regrafting of defatting fenestrated full-thickness skin by non-resection in 8 cases, and skin grafting with transfer of myocutaneous flap in 5 cases. Among them, there were 11 cases of bone and articular fixation or repair, 4 cases of principal vessels repair. All of the cases were evaluated clinically and followed up for 6 months to 3 years. RESULTS: In the 8 cases repaired by regrafting of defatting fenestrated full-thickness skin, only one case of skin necrosis, 5 cm x 2 cm in size, recovered after skin grafting; the others healed well. All of the patients recovered normal life and had normal limbs. CONCLUSION: It's crucial to make a careful assessment about the injury severity of CIDI, to stress on importance of management of both CIDI and deep injury, and to choose proper options after comprehensive assessment of the injury. PMID- 12569694 TI - [Application of autogenous bone grafting with vascular anastomosis in treatment of nonunion of fracture and bone defect]. AB - OBJECTIVE: To present a surgical choice for nonunion and bone defect. METHODS: From November 1994 to October 1997, 17 cases of nonunion of fracture and massive bone defect were treated by autogenous iliac bone and fibular bone with vascular anastomosis. Of 17 cases, there were 10 cases of nonunion of bone fracture, 7 cases of bone defect following tumor resection (4 cases of benign and 3 cases of malignant). Autogenous fibular bone grafting with vascular anastomosis, ranging from 12 cm to 29 cm in length, were employed in 12 cases; autogenous iliac bone grafting, ranging from 7 cm x 3 cm to 9 cm x 5 cm in size, were utilized in the other 5 cases. All of 17 cases were followed up for 10 months to 5 years, 3 years and 7 months in average, and were evaluated from clinical manifestation. RESULTS: Bone union was achieved in 10 cases of nonunion of fracture after bone grafting, bony refilling of the bone defect was observed in 4 cases due to benign tumor and 1 case due to malignant tumor. The other 2 cases of malignant tumor died from lung metastasis of the tumor 10 months and 12 months after bone grafting, respectively. CONCLUSION: Bone grafting is an effective surgical option in treatment of nonunion of fracture and bone defect. PMID- 12569695 TI - [Surgical reconstruction of the stability of lower cervical spine in children]. AB - OBJECTIVE: To investigate surgical reconstruction of stability of lower cervical spine in children suffering trauma, tuberculosis and tumor. METHODS: From January 1998 to September 2001, 8 cases of unstable lower cervical spine were treated by operations, of anterior decompression, massive iliac bone grafting, posterior fixation with spinous process tension band wiring, and fusion with heterogeneous iliac bone grafting. RESULTS: With an average following up of 1 year and 9 months (6 months to 4 years and 3 months), 3 cases recovered excellently, 4 cases recovered well and 1 case died of pulmonary infection. CONCLUSION: The above results indicate that anterior decompression, massive iliac bone grafting, posterior fixation with spinous process tension band wiring and fusion with heterogeneous iliac bone grafting can be used as one of the methods to reconstruct the stability of lower cervical spine in children. PMID- 12569696 TI - [Surgical management of vascular injury of extremities]. AB - OBJECTIVE: To summarize the importance of surgical management to repair vascular injury in limbs salvage, and to analyze the influence factors in the management. METHODS: From 1993 to 2000, 42 cases of 58 vascular injuries were reviewed; there were 37 males and 5 females, aging from 12 to 70 years old. Emergency operations were performed in 38 cases and selective operations in 4 cases from 1 hour to 45 days after injury. There were 22 cases of complete rupture in 32 blood vessels, 5 cases of partial rupture in 6 blood vessels, and 15 cases of vascular defect in 20 blood vessels, with 5 cm to 10 cm defect. The operation management included end-to-end anastomosis in 22 cases, side-to-end anastomosis in 1 case, vascular repair in 5 cases and vascular grafting in 14 cases. All of the cases were followed up for 6 months to 7 years. RESULTS: In those received emergency operations, it was successful in 35 cases, with amputation in the other 3 cases; after operation, there were 5 cases of post-operative angio-crisis, 1 case of hematoma and 1 case of pseudoaneurysm. In those received selective operation, all succeeded but 1 case of post-operative angio-crisis. After the follow-up, except for 3 cases of amputation, the other limbs survived; and function of the survived limbs recovered satisfactorily after operation except poor recovery in 7 cases of replantation of the limbs. CONCLUSION: To repair vascular injury immediately, to manage angio-crisis and to remove influence factors is the key to save the injured limbs and to maintain the function of them. PMID- 12569697 TI - [Diagnosis and treatment of subcutaneous cavernous hemangioma with color Doppler ultrasonography]. AB - OBJECTIVE: To discuss clinical application of the color Doppler ultrasonography in diagnosis and treatment of cavernous hemangioma in deep subcutaneous tissue. METHODS: From 1996, 15 cases of cavernous hemangioma were diagnosed and located with color Doppler ultrasonography and were embolized under monitoring of the ultrasonography or resected by operation before re-examination of the hemangioma via the color Doppler ultrasonography after the intervention. RESULTS: Direct embolization was achieved in 10 cases after pinpoint location of the hemangioma by the ultrasonography, and guided embolization was performed successfully in 2 cases via the monitoring of ultrasonography, and operation had to be adopted to remove the focus. No reoccurrence of the hemangioma was observed in all the cases. CONCLUSION: Cavernous hemangioma in deep subcutaneous tissue could be easily diagnosed and located with color Doppler ultrasonography, and could be removed by embolization under monitoring of the ultrasonography successfully. PMID- 12569698 TI - [Culture of human fibroblasts transfected by human telomerase reverse transcriptase eucaryotic expression plasmid pGRN145 and their biological characteristics in vitro]. AB - OBJECTIVE: To analysis the biological characteristics of human fibroblasts transfected by human telomerase reverse transcriptase (hTERT) eucaryotic expression plasmid pGRN145. METHODS: Fibroblasts from children's foreskin were isolated and cultured in vitro, and the fibroblasts were transfected by pGRN145 with Lipofec-tAMINE PLUS Reagent. After strict screening of hygromycin B, the positive clones were subcultured. The telomerase activity was detected by RT-PCR and TRAP-PCR technique. The cell generation cycle and apoptosis rate were detected by flow cytometry to investigate the proliferative characteristics after transfection, and the chromosome karyotype of transformed cells was analyzed. The collagen secreted by transformed cells was detected by immunohistochemical staining. RESULTS: The morphological properties of fibroblasts did not change obviously after transfection. There were telomerase activity in transfected fibroblasts, while it could not be detected in pre-transfection fibroblasts. The cell generation cycle had no obvious changes between pre-transfection and post transfection. However, the apoptosis rate of transfected fibroblasts were decreased compared with that of pre-transfection. The fibroblasts transfected by pGRN145 maintained the normal diploid karyotype, as well as the cells could normally secret type I and III collagen. CONCLUSION: The human fibroblasts transfected by pGRN145 has telomerase activity with prolonged life span of culture, which preliminarily proves the availability of establishing standard seeding cell lines of tissue engineering by hTERT plasmid transfection techniques. PMID- 12569699 TI - [Osteogenic potential of fibroblasts with reconstituted telomerase activity]. AB - OBJECTIVE: To prevent the senescence of 'seed cells' for tissue engineering, the life span of human fibroblasts is extended by reconstitution of telomerase activity, and the osteogenic potential of these fibroblasts are tested. METHODS: The pGRN145 plasmids encoding human telomerase reverse transcriptase (hTERT) were introduced into the normal human primary fibroblasts by electroporation. Telomerase activity was analyzed by TRAP-PCR assay. The beta-galactosidase stain was used to indicate the signs of cell senescence. The hTERT positive fibroblasts were then induced to form bone nodules. The bone nodules were stained by tetracycline and Alizarin Red S. RESULTS: Stable telomerase activity could be detected in the transfected fibroblasts and no signs of cell senescence were found in the fibroblasts cultured for more than 50 doublings. The hTERT positive fibroblasts could form bone nodules when they were cultured in vitro induced by bone morphogentic protein 2 and tumor necrosis factor-alpha. CONCLUSION: The fibroblasts with reconstituted telomerase activity reserve their osteogenic potential. PMID- 12569700 TI - [Animal model of immunologic change detection in vivo after xenotransplantation]. AB - OBJECTIVE: To review the current condition, test method and progress of the animal model of xeno-graft-versus-host disease(xeno-GVHD). METHODS: The literature review and comprehensive analysis methods were used in this article. RESULTS: Implanted immunologic cells, the recepient had the chance of showing host-versus-graft reaction, GVHD or microchimerism. Now, xeno-GVHD could be induced in vivo at small and large animals, it also could be supervised through many ways. CONCLUSION: Chimeric cells is very important to xeno-GVHD animal model. With this model, we can really mimic the immunologic change in vivo after xenotransplantation. PMID- 12569701 TI - [Orthopaedic applications for biodegradable and absorbable internal fixation of fractures]. AB - OBJECTIVE: To illustrate the effect and complication of orthopedic applications for biodegradable and absorbable internal fixation of fractures, and to indicate the existent problem and research aspect currently. METHODS: The recent literatures on orthopedic applications and study of biodegradable and absorbable internal fixation for fractures were reviewed. The effect of biodegradable materials on bone healing was summarized. RESULTS: It is good for the stability of fracture fixation and result of treatment. The biodegradable and absorbable internal fixation fractures had no adverse effect on bone healing. CONCLUSION: There will be more widespread application for biodegradable and absorbable materials in orthopedics, but the intensive research should be carried out to prevent its complication. PMID- 12569702 TI - [Physician for life]. PMID- 12569703 TI - [Nobel Prize in physiology or medicine 2002]. PMID- 12569704 TI - [Health care cooperation from Porto to Petrozavodsk]. PMID- 12569705 TI - [Outbreak of campylobacteriosis among soccer players]. AB - BACKGROUND: Last year Rosenborg Ballklub won the Norwegian Premier League Championship in a nerve-racking match against Lillestrom Sportsklubb. Lillestrom ended the season in 2nd place. The results may possibly have been caused by the cancellation of a match scheduled between Lillestrom and Odd Grenland. MATERIAL AND METHODS: This article describes the problem and consequences that can occur when 14 players on a Premier League team contract diarrhoea and present with vomiting either before or during a football match. RESULTS: With Campylobacter infection jejuni amply documented, the Norwegian Football Union found it necessary to cancel the match between the two Premier League teams, a decision that led to heated discussion in the media. A common opinion was that an intestinal infection, even one that affected so many players, was insufficient reason to cancel a match. INTERPRETATION: It is always difficult to foresee the consequences of continued physical activity by those infected with intestinal bacteria. It is difficult to know at what point the footballer's health may be a risk either during or after infection. PMID- 12569706 TI - [Stormorken's syndrome]. AB - In 1985, a new syndrome with the following characteristics was described: thrombopathia, thrombopenia, asplenia, miosis, headache, ichthyosis, dyslexia, muscle defect, and subsequently also hypocalcaemia. Skin and deep bleedings, leg spasms, disturbed dark vision and dyslexia are main worries. This paper describes these patients with a review of the investigations performed. Causes of the bleeding tendency are complex disturbances of the platelet membrane causing insufficient stability of the haemostatic plug, the nature of which is unresolved, but involves membrane scrambling. The muscle defect consists in tubular aggregates and high blood values of creatine kinase. A connection with the hypocalcaemia is possible, because increasing the ionic Ca with calcitriol significantly improves muscle function. Miosis is resistant to mydriatics and causes decreased dark vision, possibly also influencing dyslexia. The asplenia has little influence on immunocompetence, and the patients have survived 300 patient years without critical infections. The gene defect has not yet been unravelled. PMID- 12569707 TI - [Valerian as a sleeping aid?]. AB - BACKGROUND: Valeriana is a herbal over-the-counter drug, in Norway mainly used for insomnia. It is estimated that in Norway only, annual sales of valeriana amount to NOK 20 m. With this considerable consumption in mind, we wanted to review the scientific basis for valeriana as a sleep aid. MATERIAL AND METHODS: Through a literature search we identified 18 experimental studies examining the effects of valeriana on human sleep. RESULTS: The majority of studies reported positive effects of valeriana on subjective sleep parameters. Objective sleep measures yielded inconsistent results. All studies covered a short period of time. Few side effects of valeriana were reported. Only two studies compared valeriana to established hypnotics. INTERPRETATION: We conclude that valeriana may have some hypnotic effect. Long-term studies are clearly lacking, as well as studies comparing valeriana to well-established drug treatments for insomnia. PMID- 12569708 TI - [Botulism in Osterdalen in 1831]. AB - The earliest report we have had of an outbreak of botulism in Norway is from 1934, of five cases caused by contaminated salted and dried ham. All patients survived. This paper describes an early outbreak of botulism in the county of Hedmark in 1831 on the basis of a previously unknown report to the county governor from district public health officer Peter Heiberg (1778-1849). Two cases were reported, one of them fatal. The contaminated food was salted and fermented trout or char. PMID- 12569709 TI - [Kristian Igelsrud and the first successful direct heart compression]. AB - In 1901, at Tromso Hospital in Norway, Kristian Igelsrud performed the first successful direct heart compression in history. The event attracted little attention, and Igelsrud did little to make sure that it did. Igelsrud's operation is one of the most important contributions by Norwegian medicine to the development of anaesthesia and resuscitation. PMID- 12569710 TI - [Methodological problems in studies of acute morbidity 1870-1900]. AB - BACKGROUND: The use in medical history of morbidity data from national health statistics raises difficult issues concerning the quality of registration practice in the late nineteenth century. MATERIAL AND METHODS: Data on morbidity have been extracted from the annual medical reports from each of the seven health districts in Telemark County, Norway from 1870 to 1900. The incidence of epidemic diseases and general mortality is calculated. RESULTS: In Skien and Kragero health districts, the occurrence of epidemic disease was higher than in the five adjacent rural districts. While illness increased in Skien during most of the period, it declined in Kragero from about 1885. Throughout the period there is a clear tendency towards conformity in the patterns of development of acute morbidity and general mortality. INTERPRETATION: Health statistics are assumed to have been influenced to some extent by inaccurate diagnostics, varying numbers of doctors and their accessibility, and dissimilarities in general educational standards and in the ability and willingness to pay for medical treatment. However, these factors alone can not account for the considerable differences in morbidity between the seven health districts. It seems reasonable to conclude that variations in acute morbidity reflect differences in living conditions in the districts in question. PMID- 12569711 TI - [Sources of information about medical equipment and instruments from earlier times]. AB - The literature in medical history often provides little information about the daily work in general practice and the kind of equipment doctors used. It is possible to obtain valuable information from the Internet and from libraries and other sources, but some of the literature is not easily accessible. Literature searches, organisations, collections and networks for medical and public health history are important sources of information. Large databases and virtual museums are easily accessible on the Internet. PMID- 12569712 TI - [The gynecologist Max Sanger and his relationship with Nina and Edvard Grieg]. AB - Hundred years ago, on 12 January 1903 the gynaecologist Max Sanger (1853-1903) died. He was one of the most well-known German gynaecologists at the end of the 19th century, and during the 20 years he practiced in Leipzig, he achieved an international reputation. His scientific work covered almost the whole field of contemporary obstetrics and gynaecology, and he is still remembered for his modification of the Caesarean operation, which substantially reduced maternal mortality. Sanger was closely linked to Norway, both professionally and personally. This paper describes his work as gynaecologist and his relationship to Norway, especially his friendship with Nina and Edvard Grieg. PMID- 12569713 TI - ["Bread from stone and spiritual uplift for illiterates"--paternalistic community health promotion 200 years ago]. AB - Some of the early pioneers of community health promotion in Norway were clergymen. One of these "potato priests", as they often are referred to, was the Rev. Niels G.A. Dahl (1778-1852) in Gulen, a poor rural district in western Norway. In a spirit of paternalism he engaged in many non-religious activities promoting the lives and health of his congregation. He engaged in vaccination, medical treatment, control of infectious diseases and provision for the poor, hired a midwife, struggled hard against heavy liquor drinking among locals and promoted primary education. This article highlights Dahl's contributions to community health promotion. Effective community health promotion depends on local knowledge, as well as local authority, influence and personal engagement. Maybe a dose of the paternalistic spirit is called for also in today's health promotion? PMID- 12569714 TI - ["While breathing, one will die"--descriptions of diseases in the writings of Carl Michael Bellman]. AB - The Swedish poet Carl Michael Bellman (1740-1795) was one of the world's great song-writers. His exuberant characters include society's outcasts, many of them portrayed in great detail, with suffering and death frequently forming the backdrop for his often joyous poetry which gives unique glimpses of the living conditions in 17th-century Stockholm. The article is based on a study of the Swedish Bellman Society's "Standardupplaga", the complete edition in 19 volumes of the work of C.M. Bellman (published 1921-2001). Bellman described many of the most important diseases of his time. The descriptions are often mere depictions of symptoms and allusions to disease. They reflect Bellman's distinctive and extremely visual narrative technique, as well as the literary and social frames of reference of his time. It is argued that his frequent descriptions of suffering and death are among the reasons for, and not contradictory to, the reception he has received as a poet of life and joy. PMID- 12569715 TI - [Leonardo da Vinci--a dyslectic genius?]. AB - Leonardo da Vinci's texts consist almost exclusively of scientific notes. Working on a book on Leonardo's art, I studied all Leonardo's published texts carefully for any new information. In some prefaces I came to suspect that Leonardo might have suffered from dyslexia. This article considers the question of whether it is possible to find indications of dyslexia in Leonardo's texts and in the accounts of his life. PMID- 12569716 TI - [Mobilization of mesenchymal infection defense]. AB - BACKGROUND: Most infections are arrested in epithelial and superficial connective tissues long before antibodies and antigen specific killer cells have been induced; i.e. before the specific defence system has had time enough to come to the rescue. Microbial substances that activate and modulate this non-specific first-line defence in and near the body surfaces may enhance disease resistance, mainly by stimulating the production of anti-microbial substances by epithelia and by local activation of tissue macrophages. MATERIAL AND METHODS: There are many different microbial substances that can activate macrophages. Beta-1,3 glucans from yeast and mushrooms are the most obvious candidates for pharmaceutical development because their chemical composition and mode of action has been clarified in great detail. RESULTS: Beta-1,3-glucans in purified form provide efficient protection of animals against infections by virus, bacteria, fungi and parasites. Such enhanced protection is obtained after injection as well as after oral or mucosal administration. Beta-1,3-glucans also counteract the toxic effects of bacterial endotoxins and enhance the body's capacity to destroy cancer cells. INTERPRETATION: Activation of non-specific immunity in epithelia and in connective tissues by purified microbial substances corresponds to early events in a natural infection process and renders animals more resistant to infections. This way to enhance resistance to microbial infections has been applied with success in animal husbandry with beta-1,3-glucans administrated orally or onto mucosal surfaces. Corresponding use in human medicine is a realistic possibility, in addition to the use of microbial immune modulators as adjuvants in mucosal vaccines. PMID- 12569717 TI - [Exotic invertebrates--a health problem?]. AB - BACKGROUND: Tropical insects and arachnids are becoming popular pets in Norway. Most animals kept are harmless, but some species might cause the need for assistance from the health service. This paper presents tropical arthropods common in captivity and some challenges they might pose to health services. MATERIALS AND METHOD: This article is mainly based on articles collected with the help of Biological Abstracts, Science Citation Index and PubMed. Also, some of the findings are based on the author's experience gained through keeping tropical arthropods for 15 years and participating in international networks. RESULTS AND INTERPRETATION: There is no evidence in the literature that the bite of tarantulas is dangerous. However, the large amounts of urticating hairs in many species can cause skin irritations and serious eye inflammations that can be difficult to treat. The scorpion species imported into the Norwegian pet trade today have no medical significance, but the increasing private import of potential dangerous species via the Internet might change this situation. Walking sticks and mantis are harmless insects that have no medical significance. Most giant millipedes are also harmless, but some species have defence fluids that can cause pathological reactions if exposed to skin, mouth/throat or eyes. PMID- 12569718 TI - [Is the use of probiotics in the treatment of infections justified?]. PMID- 12569719 TI - [Magic numbers--magic and risk as explanation of disease and death]. PMID- 12569720 TI - [Physicians and sickness certification for insurance claims]. PMID- 12569722 TI - [Euthanasia in the Netherlands]. PMID- 12569721 TI - [Contraceptive injection Depo-Provera, effect on bones and teenage girls]. PMID- 12569723 TI - [Phenomenology, medicine and functional somatic disorders]. PMID- 12569724 TI - [All information about preparations is on the Medical Products Agency's web site]. PMID- 12569725 TI - [Gynecologists misuse psychiatric diagnoses]. PMID- 12569726 TI - [No effect of vitamin E in macular degeneration]. PMID- 12569727 TI - [Is Subutex a magic drug?]. PMID- 12569728 TI - Saving insurance for the sickest. PMID- 12569729 TI - A way to increase access. PMID- 12569730 TI - The view from the trenches. PMID- 12569731 TI - Liability awards and nurses. PMID- 12569732 TI - Community health. Head above water. PMID- 12569733 TI - Community health. Part and parcel. PMID- 12569734 TI - Supply chain. E-gads. PMID- 12569735 TI - Women's health. The whole woman. PMID- 12569736 TI - Medical errors. Going paperless. PMID- 12569737 TI - Rural hospitals. SHIPs come in. PMID- 12569738 TI - Cartoon therapy. Not-so-loony tunes. PMID- 12569739 TI - 1Q[3a]. Should doctors be paid for computer consultations? PMID- 12569740 TI - The big tent. PMID- 12569741 TI - The IT culture. How's yours coming? AB - Sure, you've invested in information technology. But do you really understand it? Is IT essential to your organization's success? Is your thinking about IT truly forward-thinking? Or do you end up with a lot of gizmos that, despite being really cool, just don't get used? Also: why health care is not like banking. PMID- 12569742 TI - A crisis in adolescent psych. AB - The number of children and adolescents needing mental health services has surged; yet state mental hospitals are closing and private for-profit psychiatric facilities are getting out of the business. Hospitals that still offer inpatient psychiatric beds are often full, with long waiting lists. So who fills the breach? The hospital emergency department. PMID- 12569743 TI - Medicare+Choice. AB - Created five years ago out of the Balanced Budget Act, Medicare+Choice was thought to be the next generation of Medicare, giving benefits to seniors and saving the government money. But enrollment has dropped and health plans, complaining of low reimbursement rates, have fled. What went wrong? Can it be fixed? PMID- 12569744 TI - Outsourcing information technology. An executive's guide to creating and managing IT contracts. AB - Many hospitals outsource one or more key areas of their IT operations, but not all are happy with the outcomes. In this special foldout, H&HN looks at what sectors hospitals should consider outsourcing and what questions to ask potential partners. PMID- 12569745 TI - Advocacy in '03: Why me? PMID- 12569746 TI - [The complete nucleotide sequences of A/Goose/Guangdong/2/96(H5N1) virus RNA segment 1-3 and 5]. AB - OBJECTIVE: To determine the nucleotide and amino acid sequences of PB2, PB1, PA and NP genes and compared them with sequences of A/HK/156/97(H5N1) virus for revealing the relationship between A/Googs/Guangdong/2/96(H5N1) and A/HK/156/97(H5N1) viruses. METHODS: Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA amplified by PCR, the productions of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method, using synthetic oligodeoxynucleotide primers. RESULTS: The lengths of A/Goose/Guangdong/2/96(H5N1) virus RNA segment 1-3 and 5 contain 2,341, 2,341, 2,233 and 1,565 nucleotides, respectively. They encode for PB2 (759 amino acids), PB1 (757 amino acids), PA (716 amino acids) and NP (498 amino acids) proteins. The homologies of amino acid sequences of PB2, PB1, PA and NP proteins between A/Goose/Guangdong/2/96 (H5N1) and A/HK/156/97 (H5N1) virus are 96.4%, 97.2%, 97.3% and 97.0%, respectively. CONCLUSION: The lengths of RNA segment 1-3 and 5 of Goose strain are 2,341, 2,341, 2,233 and 1,565 nucleotides, respectively. The nucleotide sequences of these genes are distinguish able from those of Hong Kong virus. PMID- 12569747 TI - [Biological characteristics of human fetal esophageal epithelial cell line immortalized by the E6 and E7 gene of HPV type 18]. AB - OBJECTIVE: Biological features including proliferation, differentiation and cell death of SHEE cell line, an immortalized epithelium of the fetal esophageal epithelium induced by HPV 18 E6E7 AAV, were studied. METHODS: SHEE cell line being cultured for more than 50 passages, were cultured in 199 growth medium and were examined by light, electron and fluorescence microscopy for growth rate, morphological features and chromosome analysis, by flow cytometry for cell proliferative dynamics, by immunohistochemistry for Ki67 and cytokeratin, and by terminal DNA transferase label (TUNEL) for apoptosis. RESULTS: At the 20th passage, the SHEE cell remained monolayer, anchorage-dependent and attachment inhibited growth. The growth curve showed proliferative phase 3-8th days, top plate phase 9-10th days and attenuative phase after 10th days. Proliferative index(PIx) 34.0%, mitotic index (MI) 2.74% (1.20%-4.80%), apoptotic index (AI) 1.30%-6.90%, chromosome analysis mainly 46 (44-54/nucleus) and DNA distribution in diploidy were calculated and described. The tonofilament expression in cell cytoplasm by electron-microscopy and positive reaction of cytokeratin by immunochemistry showed differentiative character of squamous epithelium. The cell apoptosis occured in the proliferative phase and especially increased in attenuative phase. CONCLUSION: Of biological behaviors, the SHEE cells are close to the basal cells of their original fetal esophageal mucosa keeping proliferative and differentiative potency. This study suggests that the cell death (including cell apoptosis) may be an important factor in studying of cell growth regulation and it may be an research area for cellular biological behaviors. PMID- 12569748 TI - [Cloning and expression of bluetongue virus VP3 protein in insect cells]. AB - OBJECTIVE: To elucidate the biological activities of VP3 protein of bluetongue virus (BTV) expressed in insect cells and to assemble, in future, the BTV core like particles. METHODS: First, full length of BTV13 L3 cDNA was constructed from 2 sequenced clones. The silver staining of the splicing region proved that the construction was correct. Then, a recombinant baculovirus expression vector pFB1BL3 was constructed to express the L3 gene in insect cells. RESULTS: BTV13 VP3 could be highly expressed in Sf-9 cells, the production could occupy up to 10%-15% of the total cell protein. Core-like particles could be observed after co expression of the VP3 and VP7 in insect cells. CONCLUSION: VP3 of BTV expressed in insect cells possesses biological activity and can be used to assemble BTV core-like particles. PMID- 12569749 TI - [DNA cloning and partial sequence analysis of EBV LMP1 gene isolated from human nasopharyngeal carcinoma cell line SUNE1]. AB - OBJECTIVE: To study the variation of EBV LMP1 encoding gene isolated from human nasopharyngeal carcinoma cell line SUNE1 in Guangdong. METHODS: The EBV LMP1 gene was amplified from the SUNE1 cell genome by PCR and then the recombinant vector was constructed by inserting the PCR fragment into pcDNA3. The encoding EBV LMP1 spanning from exon1 to exon3 in recombinant vector was sequenced by the Sequence Analyzer. RESULTS: The 1,312 bp encoding EBV LMP1 pieces in 2.6 kb PCR fragments were compared with the same EBV segments in B95-8 cell line. The data indicated that the rate of nucleotide sequence homology between the two fragments is 98.5% and the rate of amino acid is 96%. The restricted enzyme site of Xho I in exon1 was deleted in SUNE1 but the 30 bp deletion at the carboxyl terminus in most Chinese NPC LMP1 gene was not present. CONCLUSION: Although the LMP1 gene derived from SUNE1 had greater tumorigenicity than that derived from B95-8 cell, the high homology rate of nucleotide and amino acid sequences between them indicated that it was not the result from the variation of certain nucleotide sites but the change in amino acid domain. PMID- 12569750 TI - [Variation of primary structure in nonstructural region 5 b of hepatitis C virus]. AB - OBJECTIVE: To analyze the sequence variation in the nonstructural region 5 b(NS5 b) of hepatitis C virus (HCV) of China. METHODS: Complementary DNA fragments from sera of 49 Chinese patients were amplified by reverse transcription nested polymerase chain reaction (RT-nested-PCR) and the products were cloned and sequenced. RESULTS: 33 clones belonged to genotype 1 b and 16 clones were genotype 2 a. Several conserved nucleotide sites were found to be different from those of Japanese isolates though they belonged to an identical genotype. Comparison of average homology among the 33 clones of genotype 1 b and the 16 clones of genotype 2 a indicated that the average homology among genotype 2 a was lower than that among genotype 1 b. A deletion of three nucleotides and a frame shift, resulting from the introduction of an in-frame stop codon, were firstly observed in the NS5 b region. CONCLUSION: The nucleotide sequences of different HCV isolates of a same subtype may possibly possess the characteristics for different geographical distribution and so a definite epidemic HCV strain may emerge from a definite area. PMID- 12569751 TI - [New subtype of coltivirus isolated from mosquitoes in the northeast part of China]. AB - OBJECTIVE: The purpose of this study was to isolate new arbovirus from mosquitoes collected from the northeast part of China during the summer of 1996. METHODS: The cell used for isolation were C6/36 and BHK-21 cells. RESULTS: Two strains (NE97-12 and NE97-31) of coltivirus were isolated among 68 mosquito specimens. The viruses caused cytopathogenic effects on C6/36 cell, but not on BHK-21 cells. They were resistant to both 5-IDU and ether, but sensitive to pH3.0 and temperature at 56 degrees C for 30 minutes. IFA of new isolates with mouse ascitic fluid anti-coltivirus TRT2 strain showed positive reaction. Polyacrylamide gel electrophoresis (PAGE) of two isolates showed 12 segments of double-stranded(ds) RNA and PAGE profiles were 6-5-1, which were different from that of coltivirus TRT2 strain(6-6). There was no cross neutralization between new isolates and TRT2 strain by tissue culture cross-neutralization test. CONCLUSION: The results indicated while new isolates were serologically related to TRT2 strain of coltivirus by IFA, the neutralization test suggests that it represents a distinct serotype. These new isolates isolated firstly from the northeast part of China belong to a new subtype of coltivirus. PMID- 12569752 TI - [Production of HBV surface antigen-specific target cell]. AB - OBJECTIVE: To analyze the hepatitis B virus surface antigen (HBsAg)-specific cytotoxic T-cell (CTL) response during hepatitis development and investigate the effect of some factors on CTL response. METHODS: The recombinant retrovirus plasmid PLXSN-S was tranferred into PA317 by electroporation and the pseudoviruses produced from PA317 were used to infect EL4. Clones which highly expressed HBsAg were selected with limited dilution. RESULTS: A clone expressing HBsAg the highest (A = 0.85 in supernatant at 48th hour) was obtained which steadly expressed HBsAg in vitro after at least 100 passages. It worked very well when used as target cells in CTL response after C57 mice immunized with PLXSN-S and recombinant adenovirus vector (rAdv-B7-2-S). CONCLUSION: We constructed HBsAg specific target cell line which steadly expressed HBsAg. It may play an important role in HBsAg CTL response and immunopathogenetic mechanisms of hepatitis B. PMID- 12569753 TI - [Detection of human papillomavirus and herpesvirus genotypes in biopsy specimens from cervical carcinoma by PCR-endonuclease cleavage]. AB - OBJECTIVE: This study was to detect the human papillomavirus (HPV) and herpes simplex virus (HSV) genotypes in biopsy specimens of cervical carcinoma and genital verruca by PCR-endonuclease cleavage. METHODS: Biopsy samples of were obtained from 390 patients with cervical carcinoma, HPV and HSV genotypes were detected by PCR and endonuclease cleavage. RESULTS: The results indicated that HPV types 16, 18, 35 were found in 152 (38.9%) of 390 cervical carcinoma, in 56 (31.8%) of 176 cervical erosion, while HPV types 6, 11 were found in 80 (58.8%) of 136 acuminate erruca. The HSV-2 type were found in 135 (34.6%) of 390 cervical carcinoma and in 33 (18.7%) of 176 cervical erosion. The HPV 16, 18, 35 types and HSV-2 type found in normal cervical secretions of the control group all were 3.2%, There was significant difference between the test group of cervical carcinoma and the control group of normal cervical secretion (P < 0.001 by x 2 analysis). The restriction endonuclease analysis of these amplified products showed that positive rates of HPV 16, 18, 35 types and HSV-2 type from cervical carcinoma were 23.3%, 14.6%, 1.0% and 34.6%, respectively. While HPV 6, 11 types from acuminate verruca were 36.0% and 22.8% positive respectively. CONCLUSION: The results suggest that HPV 16, 18 types and HSV-2 type are associated with majority of the cervical carcinoma. PMID- 12569754 TI - [Effect of dengue virus infection on the production of ET-1 and PGI2 by human vascular endothelial cells]. AB - OBJECTIVE: To study the effect of dengue virus infection on the production of endothelin 1(ET-1) and prostacyclin 2(PGI2) by human umbilical cord vein endothelial cells (HUVECs). METHODS: HUVECs were infected by dengue-2 virus (DV 2), the culture supernatants were collected at 4, 24, 48, 72 and 96 hours postinfection, and the levels of ET-1 and PGI2 were measured by radio immunoassay. RESULTS: The results showed that inhibition of production of ET-1 and PGI2 was induced by dengue virus infection in HUVECs. Marked inhibition of ET 1 and PGI2 production was observed as early as 4 h postinfection and the inhibitory effect continued until 96 h postinfection. Significant differences were shown between the ET-1 concentration of the virus-infected and non-infected cells. Nevertheless, the inhibitory effect of dengue virus infection on the production of PGI2 gradually decreased and the PGI2 concentration reverted to normal level at 96 h postinfection. No significant difference between the PGI2 concentration of the virus-infected and non-infected cells was seem. CONCLUSION: The present study suggests that dengue virus infection of vascular endothelial cells could affect their normal functions of secretion of vasoactive substances, resulting in increased vascular permeability and impairment of homostasis and blood coagulation. Therefore, functional impairment of vascular endothelial cells induced by dengue virus might be an important aspect in the pathogenesis of DHF/DSS. PMID- 12569755 TI - [Analysis of hepatitis C virus genotypes in Guizhou area, using second generation line probe assay]. AB - OBJECTIVE: To analyse the frequency of various hepatitis C virus genotypes in different populations with HCV infection from Guizhou area. METHODS: Ninty-eight HCV isolates obtained by using RT-nested PCR from 37 patients with chronic hepatitis C (PHCs), 15 patients with hemopathy (PHs), 12 blood donors (BDs) and 34 intravenous drug-users (IVDUs) were genotyped by using the second generation line probe assay. RESULTS: Among 37 HCV isolates from PHCs, there were 34 (91.9%) genotype 1b, 3 (8.1%) mixted genotypes (1 for 1a + 1b, 1 for 1b + 2, 1 for 1a + 1b + 2a/2c, respectively). Among 15 PHs, 14 (93.3%) were type 1b, 1(6.7%) were type 1b + 2, among 12 BDs, all(100.0%) were type 1b. Among 34 IVDUs, 13(38.2%) were type 1b, 10(29.4%) were type 6a, 1(2.9%) was type 3a, 4(11.8%) were type 3b and 6(17.6%) were mixed types(2 for 1a + 2, 1 for 1a, 1b + 2, 1 for 1b + 3b, 1 for 3b + 2, 1 for 1b + 6a, respectively). CONCLUSION: The results showed that HCV subtypes 1a, 1b, 2a/2c, 3a, 3b and 6a belonging to 4 HCV genotypes(1,2,3,6) were found from different populations in Guizhou, of these, 3a, 3b and 6a were only found in IVDUs and their total frequency was above 50%. PMID- 12569756 TI - [A sero-epidemiological study of HCV and HBV infection among intravenous drug abusers]. AB - OBJECTIVE: To investigate hepatitis B and C virus infection in drug addicts in Guangzhou. METHODS: Serum samples from drug users and normal control people were collected. Serum antibodies against hepatitis C virus (HCV) and hepatitis B virus (HBV) were detected by ELISA. RESULTS: The results showed that hepatitis prevalence is significantly higher among the drug users than that in normal control group (for HCV 67.49% to 3.75%, for HBV 90.48% to 55.63%, and for the mixed infections 62.73% to 3.75%). CONCLUSION: In Guangzhou, the very high infection rates of HBV and HCV were found in drug addicts. Our results emphasize intravenous drug use is the major risk factor for HBV and HCV infections. PMID- 12569757 TI - [Hepatitis B virus core promoter mutations in patients with fulminant hepatitis]. AB - OBJECTIVE: To detect hepatitis B virus core promoter (CP) mutations in patients with fulminant hepatitis. METHODS: Polymerase chain reaction amplified serum HBV DNA fragments were directly sequenced. RESULTS: There were 2-12 nucleotide substitutions in CP region in the 7 subacute fulminant hepatitis patients studied. An 11 bp nucleotides insertion was found in one patient. Mutations in CP were usually seen in the first and the second A T rich regions. The A to T mutation at nt 1,762 and G to A mutation at nt 1,764 were found in 4 cases, 3 of them were HBeAg negative. The third A T rich region was kept intact in all the 7 patients, so did the initial site of HBV replication (DR1) and the initial site of mRNA transcription (1,783/1784 or 1,790 +/- 1 for precore mRNA and 1818 for pregenome-C/P mRNA). CONCLUSION: CP mutations in patients with fulminant hepatitis are common, most of the CP variations occur in the first and the second A T rich regions, and these mutations may impede the transcription of precore mRNA and affect the expression of HBeAg. PMID- 12569758 TI - [Virus interaction in HBV and HCV super infection]. AB - OBJECTIVE: To investigate virus interaction in HBV and HCV superinfection. METHODS: The changes of serum markers of 30 patients with HBV and HCV superinfetion were observed. Meanwhile, mutation of HBV precore region at nt 1,896 and the amount of TNF alpha and IL-6 in sera were also investigated. RESULTS: Compared with HBV or HCV single infection, positive rates of HBeAg, HBV DNA and HCV RNA in superinfectim were lower, positive rates of anti-HBe was higher, the average titers of HBsAg, anti-HBcIgG and anti-HCV were significantly lower, too. In some patients, HBsAg were seronegatively convested. Occurrence rate of HBV precore region mutation at nt1896 and levels of TNF alpha and IL-6 in sera were higher in super infection than those in single infection. CONCLUSION: Interference existed between HBV and HCV superinfection, conversion from HBeAg to anti-HBe was not only due to the suppression by HCV but also to the mutation of HBV precore region, and HCV may be a reason causing HBV precore region mutation through increasing the pressure of the body's immunosystem. PMID- 12569759 TI - [The feature of influenza virus activity and genetic characterization of HA1 gene of influenza A H3N2 viruses isolated from 1994-1997 in Shenzhen]. AB - OBJECTIVE: To understand the feature of influenza virus activity and genetic characterization of HA1 gene of influenza A H3N2 viruses isolated in recently years in Shenzhen. METHODS: The specimens were inoculated into amniotic and allantoic cavities of the embryonated chicken eggs for viral isolation. The isolates were typed with HI test. Virion RNA extracted from allantoic fluid by Rneasy RNA extraction kit was used for RT-PCR amplification. Afterward, the product of PCR was purified with purification kit. RNA sequence analysis was performed by Sanger dideoxy-mediated nucleotide chain termination method. RESULTS: Influenza A H3N2, H1N1 and B viruses were cocirculating in men in Shenzhen in recent years. However, the extent of their activities was not the same in a single year. The influenza B virus was the predominant strain in 1994 and 1997, but the predominant strain was H3N2 virus in 1996. The genetic analysis showed that the substitution of amino acids occured year by year and was mainly located at antigenic determinants or receptor binding site on HA1 domain of H3N2 viruses. CONCLUSION: Influenza A H3N2, H1N1 and B viruses were still co circulating in men in Shenzhen in recent years, the predominant strain of influenza virus was different in different years. An increase of influenza A H3N2 virus activity in 1996 was due to the substitution of amino acids sequences on viral HA1 domain. PMID- 12569760 TI - [The study on clinical significance of human herpesvius 6 infection of NPC tissues]. AB - OBJECTIVE: To study the infection of human herpesvirus 6(HHV-6) in NPC and its role in the carcinogensis of NPC. METHODS: By using polymerase chain reaction (PCR) and in situ hybridization (ISH) HHV-6 and EBV DNA to from the paraffin embedded tissues of normal nasopharynx precancerous nasopharynx and nasopharyngeal carcinoma. EBV LMP1 was also detected in 36 tissues of NPC by immunohistochemistry (IHC). RESULTS: HHV-6 DNA PCR were detected in 30.9% (13/42) of tissues of NPC, 4.7% (1/21) of precancerous nasopharynx and non in the normal nasopharynx. Within the same tissues, EBV DNA positive rates were 95.2% (40/42), 66.6% (14/21) and 25.9% (7/27), respectively. By using ISH, HHV-6 DNA were detected only in 11 NPC tissues out of 13 HHV-6 DNA positive NPC cases and EBV DNA were detected in 47.6% (10/21) of precancerous nasopharynx tissues and 85.7% (36/42) of NPC cases. EBV LMP1 expression were detected in 47.2% (17/36) of NPC tissues. CONCLUSION: NPC tissues can be infected with both HHV-6 and EBV, HHV-6 infection is correlated with the expression of EBV LMP1, suggesting that HHV-6 plays a direct/or indirect role in carcinogenesis of NPC via upregulating the expression of EBV LMP1. PMID- 12569761 TI - [The treatment effect of ribavirin to hemorrhagic fever with renal syndrome on the kinetics of serum creatine phosphate kinase isoenzyme]. AB - OBJECTIVE: To study the effect of ribavirin treatment on kinetics of serum creatine phosphate kinase isoenzyme(CK-MB) of patients with hemoarrhagic fever with renal syndrome(HFRS). METHODS: CK-MB levels in 155 HFRS patients treated with larger dose of ribavirin and with routine therapeutic dose were observed dynamicaly, using randomly controlled double-blind method. In addition, CK-MB levels in different disease phases and different clinical types of HFRS were compared. RESULTS: The results showed that CK-MB levels were significantly increased in febrile phase and decreased gradually to normal during convalescent phase in both groups, but in febrile phase, CK-MB levels in ribavirin-treated group and control group were 55.25 +/- 18.25 IU/L and 69.63 +/- 23.52 IU/L, respectively, the treated group had a lower level (P < 0.05). The severe and critical group treated by ribavirin showed a lower CK-MB level of 47.74 +/- 24.82 IU/L, while the control group showed a higher level of 66.08 +/- 25.01 IU/L(P < 0.05). CONCLUSION: These results indicated that an early application of large dose of ribavirin in treatment of HFRS can reduce the serum CK-MB levels in patients with HFRS. PMID- 12569762 TI - [Influence of IL-4 on the expression of TNF-alpha and IL-1 alpha mRNA by PBMCs from patients with subfulminant viral hepatitis before and after treatment]. AB - OBJECTIVE: The aim of this study was to compare the influence of IL-4 on the expression of TNF-alpha and IL-1 alpha mRNA by PBMCs from patients with subfulminant viral hepatitis(SFH) before and after IL-4 treatment. METHODS: The expression levels of TNF-alpha and IL-1 alpha mRNA by PBMCs were assessed by semiquantitative RT-PCR. RESULTS: IL-4 suppressed the expression of both TNF alpha and IL-1 alpha mRNA by PBMCs from SFH patients before and after treatment dose-dependently. However, their dose-response curves showed significant differences. The inhibitory effect could be observed at a concentration of 100 U/ml and was near to maximum at 1,000 U/ml in acute phase, whereas the same suppressive action of IL-4 was reached at 100 U/ml in recovery phase. When PBMCs were cultured in the presence of IL-4 at a concentration of 100 U/ml, IL-4 down regulated the production of both TNF-alpha and IL-1 alpha mRNA by acute phase PBMCs only to 18.18-21.98% of control cells(in absence of IL-4), while the inhibitory rates were all near to 50% in recovery phases. Moreover, it was also found that the suppressive effect of IL-4 on the expression of PBMCs TNF-alpha and IL-1 alpha mRNA in the patients with endotoxinemia and HBeAg was significantly decreased in acute phase. CONCLUSIONS: The inhibitory effect of IL 4 on the expression of PBMCs TNF-alpha and IL-1 alpha mRNA is markedly decreased in acute phase as compared with that in recovery phase and this declined response may be related to endotoxinemia and viremia. PMID- 12569763 TI - [An inquire into the infection of a new virus(TTV)]. AB - OBJECTIVE: The aim of this study is to find out the prevalence of TTV in the cryptogenic hepatitis' and the clinical features of TTV infected patients. METHODS: TTV DNA in the serum was tested by nested PCR and the clinical data was analyzed. RESULTS: Infection rate of the TTV in cryptogenic hepatitis was 38.6%. It was a sporadic disease, and involved mainly the adults, with no sexual difference. The symptoms were notable, often with jaundice, but the level of transaminase were elevated moderately. Histology revealed the portal area inflammation. If accompanied with other disease, the illness would be exacerbated. CONCLUSION: TTV would be accounted for part of the reason in patients with cryptogenic hepatitis. PMID- 12569764 TI - [The significance of TT virus detection in persons with elevated alanine aminotransferase during routine medical check up and patients with liver diseases]. AB - OBJECTIVE: To investigate the significance of the new DNA virus (TTV) infection in persons with elevated alanine aminotransferase (ALT) during routine medical check up and patients with liver diseases, and analyze its pathogenesis. METHODS: PCR was used in the detection of TTV DNA in 19 serum samples from ALT elevated persons and 41 from ALT normal persons during their routine medical check up, and 182 serum samples from patients with various kinds of liver diseases. PCR products were confirmed by RFLP. Other hepatitis viruses were simultaneously detected using routine methods. RESULTS: TTV DNA was detected in 10.53% (2/19) and 2.44% (1/41) in groups of ALT elevated and control respectively with no significant difference between their detective rates (P > 0.05). There were 9 cases with current HBV infection, 1 case with hepatitis C and 1 case with hepatitis A in group of elevated ALT. Two TTV positive cases were both superinfected with HBV. The detecting rate of TTV DNA in patients with liver diseases was 4.4%. Only one case out of 18 non A to G hepatitis showed TTV DNA in serum. CONCLUSION: There were TTV infections in Chinese population. The main cause for ALT elevation in persons during routine medical check up was HBV infection. There were TTV infections in patients with liver diseases too. However, TTV wasn't the main cause of non A to G hepatitis. Thus, the pathogenesis of TTV might be very weak. PMID- 12569765 TI - [Establishment of hepatitis C virus reproductive model of human peripheral blood mononuclear cells transformed by Epstein-Barr virus]. AB - OBJECTIVE: For the present, one of the crucial questions to study pathogenic mechanism of hepatitis C and its drug therapy is to seek a suitable cell model which may support the replication of hepatitis C virus(HCV). In this article, we made effort on the study of this aspect. METHODS: Using Epstein-Barr virus(EBV), we transformed the peripheral blood mononuclear cells(PBMC) isolated from a patient diagnosed as chronic hepatitis C. HCV RNA from the cultured cells and the supernatants were analysed by reverse transcriptase-polymerase chain reaction(RT PCR) at an intervals of one month. RESULTS: HCV positive-strand RNA could be detected continually in the cultured cells for more than one year since the establishment of the transformed PBMC clone whereas the negative-strand RNA were also observed in the cultured cells. Interestingly, the HCV positive-strand RNA could be identified in supernatants intermittently. CONCLUSION: HCV may exist in the cultured cell line for a longer period and reproduce in and secrete out of the cells. This will offer a new cell model for studying the molecular biology and immunology of HCV. PMID- 12569766 TI - [Detection of serum antibody of coxsackievirus A24 variant]. AB - OBJECTIVE: To observe the serum antibody in residents after outbreak of acute hemorrhagic conjunctivitis (AHC) caused by coxrackievirus (CA24 v). METHODS: The serum was diluted in 1:10 and mixed with 100 TCID50 CA24 v in volume. The same the mixture was incubated 1 hour in 37 degrees C and then observed for cytopathic changes of the infected HeLa cells. RESULTS: The post-epidemic antibody positive rate in residents was 49.67% and in 19-25 age group was about 69.49%. It had significant difference statistically (P < 0.01). CONCLUSION: The raising of CA24 v antibody level in residents was relevent with age. May be it was caused by the opporturnities of taking part in social activities in different age groups. PMID- 12569767 TI - [Detection of cytomegalovirus antigens and virus DNA in the peripheral blood after organ transplantation]. AB - OBJECTIVE: To diagnose timely the cytomegalovirus(CMV) infection of recipients after organ transplantation. METHODS: Thirty two organ transplanted recipients were demonstrated for CMV infection based on detection of antigenemia by immunoperoxidase assay, viremia with virus isolation, DNAemia by polymerase chain reaction (PCR) and in situ hybridization (ISH) in peripheral blood samples. RESULTS: Of 89 blood specimens examined, 35(39.3%) were positive for antigenemia, 25(28.1%) for viremia, 37(41.6%) for DNAemia by ISH, and 51(57.3%) for DNAemia by PCR. The results indicated that DNAemia assay with ISH and PCR and antigenemia assay were suitable methods for rapid, early, and sensitive detection of CMV infection. The results correlated with clinical symptoms. CONCLUSION: We suggest that detection of antigenemia, viremia, and DNAemia can monitor CMV infection of organ transplaned recipients and provide indication to antiviral therapy. PMID- 12569768 TI - [The therapeutic effect of Chinese medicinal herbs on HIV patients]. AB - OBJECTIVE: To study the therapeutic effect of Chinese medicinal herbs on HIV patients. METHODS: We adopted the combined Chinese medicinal herbs for treatment, its chief component is to enhance human immunity and the complementary part is suited for lightening the clinical synptoms. The clinical aspects, the CD4, CD8 lymphocyte numbers and the plasma viral load of the patients were compared before and after the treatment. RESULTS: Of 28 patients exemined, they all had different degrees of body weight increasing, averagely increased 5.4 kg after treatment. 7 cases having a long-time fever, 4 cases of diarrhea, 2 cases of large scale skin ulcer and 1 case of skin rash all had their synptoms disappeared ofter one month treatment. 10 cases, who had lymph nodes enlargement, after treated five months with Chinese medicinal herbs, 3 of them the swelling lymph nodes disappeared, 7 of them the lymph nodes diminished or decreased in number. CD4 lymphocytes increased in 42.9% and 50.0% patients and viral load decreased in 71.4% and 80.0% patients after two months and five months treatment with Chinese medicinal herbs respectively. Taking clinical symptoms and laboratory indexs into consideration, the total effective rate of Chinese medicinal herbs reached 94.0%. CONCLUSION: The Chinese medicinal herbs can obviously improve the clinical symptoms, increase CD4 lymphocytes and decrease serum viral load of the HIV patients, thus provides evidence that the Chinese medicinal herbs can be used for control of HIV infection and treatment of AIDS patients. PMID- 12569769 TI - [A report of recombinant interferon-alpha therapy in 19 patients with acute hepatitis B]. AB - OBJECTIVE: In patients whose HBsAg are still positive after 8 weeks of acute hepatitis B attack, the chronic rate will increase. We try to treat them with IFN for reducing the rate of chronicity. METHODS: 19 patients were injected with rIFN alpha 3 MU, q.o.d. for 12 weeks(treatment group), those who didn't have anti-HBs response after treatment were vaccinated with hepatitis B vaccine(30 micrograms, i.m., once weekly for 3 weeks); 15 patients received common therapy except rIFN alpha(control group). RESULTS: HBsAg negative seroconversion rate was 95.0% (18/19) in treatment group and 53.0% (8/15) in control group(P < 0.05). No one responsed for anti-HBs in 18 HBsAg negatively converted patients in treatment group, but in control group 87.5% of HBsAg negatively converted patients developed anti-HBs (P < 0.001). Vaccinating the patients of treatment group, who didn't developed anti-HBs, with hepatitis B vaccine, 94.0% of them developed anti HBs. After 24 to 240 week follow-up, no one relapsed in treatment group, in control group 8 HBsAg negative patients didn't replase, 6 out of 7 HBsAg positive patients were still positive at the termination and follow-up. CONCLUSION: The therapy with rIFN-alpha can reduce the chronic rate in the patients with acute HBV infection. PMID- 12569770 TI - [Progress in the study of formation of virus capsid and nucleic acid packaging]. PMID- 12569771 TI - [Discovery of men infected by avian influenza A (H9N2) virus]. AB - OBJECTIVE: To understand whether the avian influenza A(H9N2) virus can infect men or not. METHODS: Seroepidemiological surveys for avian (H9N2) virus in human, chickens and pigs were conducted. The specimens for viral isolation were taken from throat of patients with influenza like disease, as well as from chickens, then the specimens were inoculated into embryonated chicken eggs. Afterward, the idsolates were identified with HI and NI tests. Meanwhile, the patients who would be studied individually were found to carry H9N2 virus. RESULTS: Approximately 19% of human had antibody to H9N2 virus with HI titers > or = 20, 5 strains of influenza A (H9N2) virus were isolated from the patients. CONCLUSION: Avian influenza A(H9N2) virus can infect men. PMID- 12569772 TI - [The identification of recombinant HIV-1 strains in IDUs in southwest and northwest China]. AB - OBJECTIVE: To search for possible recombination of HIV-1 strains in China. METHODS: The blood samples were collected from the HIV infected individuals in areas where epidemic of more than two subtypes of HIV-1 were found. The HIV env and tat genes were amplified by nested PCR from PBMCs and directly sequenced and analyzed. RESULTS: The sequence analysis were performed on 14 HIV-1 strains collected in the subtype B' and C epidemic areas of China. No evidence of recombination after sequencing of env was found. The recombnant HIV-1 strains of subtype B' and C were found in 10 of the 14 samples tested when the first exon of the tat gene was sequenced. In addition, three subtype B' and one subtype C non recombinant HIV-1 strains were found among IDUs in Sichuan Province. CONCLUSIONS: The recombinant HIV-1 strains of subtype B' and C were firstly identified in Sichuan Province of southwest China and Xinjiang Autonomous Region of western China. Their identical sequences and recombinant patterns indicated the same origin of the recombinant strains, it strongly indicates a close correlation between these two HIV epidemic regions. Since no other subtype of HIV-1 except the recombinant was found in Xinjiang, whereas both subtype B and C as well as B'/C recombinant were found in Sichuan, the recombination was most probably happened in Sichuan Province rather than in Xinjiang. PMID- 12569773 TI - [Expression of recombinant HIV-1 envelope glycoprotein gp41]. AB - OBJECTIVE: To construct and express HIV-1 env gp41 gene for develoing a simple and rapid test for HIV-1 infection. METHODS: HIV-1 env gp41 gene of BH10 strain (nt6,977-7,497) was constructed into expressing vector pBV221 and expressed in E. Coli HB101. The expressed proteins were purified on 15% SDS-PAGE, the specific protein gel was cut down, transferred onto nitrocellulose membrane and stained with ponceau for 10 minutes. The membrane was detected with positive and negative serum respectively. The membrane was blocked with blocking buffer and cut into 2 mm of each strip and fixed into the well of thin plastic plate. RESULTS: We obtained a strand plasmid expressing HIV-1 env gene and the protein. CONCLUSION: The results showed that: (1) HIV-1 env gp41 protein can be used to detect HIV-1 antibody in serum of individual; (2) The expressed protein is a nonfusion protein and has high specificity and sensitivity to HIV-1. PMID- 12569774 TI - [Expression of non-structural region 3 gene of the Chinese HGY and analysis of the antigenicity of the recombinant proteins]. AB - OBJECTIVE: This study is to analyze the antigenicity of the NS3 proteins of Chinese HGV and their potential use in the serological diagnosis. METHODS: All three gene fragments of NS3 region of Chinese HGV were cloned into the pRSET vectors to construct recombinant plasmids. In E. coli BL21, all three recombinant plasmids achieved a high expression level with induction of IPTG. The expressed products were analyzed with Western blot and ELISA. RESULTS: The recombinant protein PA, P3 and P4 have a molecular weight of 42,000, 30,000 and 24,000, respectively. They all could react with HGV positive sera in Western blot and ELISA. Among them, the protein that covers the N terminal of NS3 region of HGV had a stronger reaction with HGV positive sera than the other two proteinsdid. CONCLUSION: The N terminal in the NS3 region of Chinese HGV includes an dominant antigenic determinant, and its gene product has relatively strong antigenicity. PMID- 12569775 TI - [Immortalization of human fetal esophageal epithelial cells induced by E6 and E7 genes of human papilloma virus 18]. AB - OBJECTIVE: For studying the relationship between HPV and esophageal carcinoma, an immortalized human fetal esophageal epithelial cell line (SHEE) was established. METHODS: The human fetal esophageal tissues were cultured and infected with HPV 18 E6E7 AAV. It was examined by light- and electron-microscope for morphological changes, by PCR and FISH for detection of HPV E6E7 and by soft agar culture and nude mice inoculatior for dectecting tumor transformation. RESULTS: The cell line has become immortal and has propagated cntinuously for more than 50 passages. After a long-term culture, the phenotype keeps the characteristics of primary epithelial cells. They showed as monolayer growth and anchorage dependent growth without forming colonies in softagar. They were nontumorigenic in nude mice. SHEE cells contained tonofilaments in its cytoplasm by electron microscopic examination and showed cytokeratin positive in immunohistochemical procedure. So it shows that the cells are squamous epithelium in origin. The cell line contained the HPV 18 E6 and E7 genes by FISH and PCR assay. CONCLUSION: Establishment of the esophageal epithelial cell line SHEE successfully immortalized with HPV 18, E6E7, supports that the HPV18 may be related to the etiology of esophageal carcinoma. It will facilitate further research on etiology and pathogenesis of esophageal carcinoma. PMID- 12569776 TI - [Expression of human prp gene in prokaryotic cells using GST fusion protein expression system]. AB - OBJECTIVE: To study the biological features of cell-surface protein PrPc, which is thought to be involved in the prion-associated diseases after converting to a proteinase-resistant isoform PrPSc posttranslationally, and to establish an effective immunologic diagnostic method using PrPc as antigen. METHODS: Amplifying and cloning the human prp gene from lymphocytes of two normal Chinese, after confirmed by DNA sequence analysis, the genes were separately subcloned into a GST-fusion expression plasmid. RESULTS: Sequence analysis showed that one contatined a point mutation that induced the 65th amino acid "Trp" inverting to a stop codon "TAG", whereas the other had the same sequence as the published standard prp gene. Both the standard and the mutated prp genes were separately subcloned into a GST fusion protein expression vector and expressed in the prokaryotic cells effectively. Western blot assay revealed that both of them expressed GST-PrP fusion proteins and could be recognized by PrP specific monoclonal antibody. CONCLUSION: It suggests that human PrP protein can be expressed in the GST fusion protein expression system and the expressed proteins hold good immune-reactivity. PMID- 12569777 TI - [Establishment of hybridomas secreting monoclonal antibodies to HIV-gp41 and HCV NS3]. AB - OBJECTIVE: To establish the monoclonal antibodies of HIV and HCV for developing serological diagnostic method. METHODS: The antigens of HIV gp41 and HCV NS3 expressed from bacteria were mixed to immunize BALB/c mice. The hybridoma cell lines were obtained by fusing SP2/0 myeloma cells with spleen cells from immunized mice. RESULTS: Four hybridoma cell lines secreting specific antibodies against gp41 of HIV and six hybridoma cell lines secreting antibodies to NS3 of HCV were obtained. ELISA sandwich method for detection of viral antigens with McAbs was developed. CONCLUSION: A quick and convenient method for obtaining McAbs was developed using mixture of different antigens for immunizition of mice. The monoclonal antibodies had high senusitivity and were stably secreted by hybridoma cell lines. This technique is worthy to be popularized. PMID- 12569778 TI - [Expression of hepatitis E virus structural gene in E. coli]. AB - OBJECTIVE: To obtain recombinant antigen for development of vaccine against hepatitis E virus. METHODS: Amplified the structural gene (5,816-7,126 nt) by PCR. The upstream primer was 5'-CCATATGAATTCAATAACCTC-3' and the downstream primer was 5'-GGGATCCTATAACTCCCGAGT-3'. Cut the PCR product with Nde I and BamHI, then inserted this fragment into the plasmid pET-11 where a cut was by the same restriction endonucleases. The expression plasmid named pEa47 was transformed into E. Coli BL21. The recombinant strains were grown at 37 degrees C and induced by IPTG. The recombinant protein was confirmed by Western blot analysis using serum from hepatitis E patient. RESULTS: The structural gene of hepatitis E virus from open reading frame 2,224-660 aa, was expressed in E. Coli BL21. Western blot assay showed that the expressed 50,000 recombinant protein specifically reacted with the serum antibody from the hepatitis E patient. CONCLUSION: The protein might be useful to develop vaccine against hepatitis E virus infection. PMID- 12569779 TI - [Influence of aluminum adjuvant to experimental rabies vaccine]. AB - OBJECTIVE: To study whether the rabies vaccine for human use should contain aluminum adjuvant. METHODS: Testing vaccine antibodies and efficacy (ED50), comparing the effect between aluminum adjuvant contained and non-aluminum adjuvant contained vaccines in a new animal model which accords with the rabies field practice. RESULTS: At fourth and seventh day after immunization, the neutralizing antibody titres of the rabies vaccine containing aluminium adjuvant were much lower than that of the vaccine not containing aluminum adjuvant. In the NIH efficacy test the ED50 of the vaccine containing aluminum adjuvant was 93-132 ng while the ED50 of the vaccine not containing aluminum adjuvant was 221 ng, but the NIH test does not accord with the rabies field practice. In that new animal model, aluminum adjuvant to rabies vaccine had not any promoting effect for preventing and treating rabies. CONCLUSION: Aluminum adjuvant to rabies vaccine has no advantages, this paper suggests that the vaccines containing and not containing aluminium adjuvant had better compare in human bodies. If the results are the same as our experiments, the aluminum adjuvant should be eliminated from rabies vaccine for human use. PMID- 12569780 TI - [Cloning of 0-17.5 mu and sequencing of 0-4.8 mu of the lefe and DNA of the Ad7 vaccine strain left end]. AB - OBJECTIVE: Cloning of 0-17.5 mu DNA fragment of adenovirus 7 vaccine strain and sequencing of 0-4.8 mu fragment (1,737 bp). METHODS: Isolating and purifying Ad7 vaccine genome from the A549 cultured cells, putting 0.3-17.5 mu fragment into pAd7T plasmid, then sequencing the 0-4.8 mu fragment including inverted terminal repeats (ITR), packaging sequence and Ela region. RESULTS: We obtained 0-17.5 mu fragment of Ad7 vaccine strain genome and sequenced its left terminal 1,737 bp. Sequence analysis showed that the Ad7 vaccine strain Ela region encodes 6,300, 24,000 and 28,000 proteins. Compared with equivalent region of Ad7 Gomen strain, they share the homology of the nucleotide sequence 98.9%, 97.3%, 97.5% and the homology of the deduced amino acid sequence 96.6%, 96.5%, and 96.9% respectively. When compared with Ad7 Grider strain, they share the homology of the nucleotide sequence 100%, 99.7%, 99.7% and the homology of the deduced amino acid sequence 100%, 99.1%, and 99.2% respectively. CONCLUSION: The Ad7 vaccine strain left terminal 1,737 bp nucleotide sequence showed a high homology with corresponding region of Ad7 Gomen strain and Grider strain. PMID- 12569781 TI - [Preliminary study on ToRCH epidemic laws in Wuhan region]. AB - OBJECTIVE: To investigate the ToRCH epidemic law in Wuhan region. METHODS: The ToRCH infection in the normal populations of Wuhan region was inspected through a whole year (from Mar. 1997 to Feb. 1998) by detecting the specific IgM antibodies to ToRCH in sera. RESULTS: The results showed that in the whole year, the average positive rates of IgM aitibodies to Toxo, RuV, CMV and HSV were 5.8%, 7.5%, 4.2% and 4.9% respectively. Three characteristics of the ToRCH epidemic in the region were concluded: (1) The epidemic of RuV obviously changed with seasons. Its infection rates were the highest (8.5%, 19.5%, 14.0%) during spring (March, April and May). However, the changes of Toxo, CMV and HSV were not obvious with seasons; (2). The adult infections were mainly recessive, which had no apparent clinical symptoms; (3) The cross infections of the four pathogens were common, the cross-reaction rate of the surveyed year was 2.6%. CONCLUSIONS: RuV infection of pregnant woman may result in congenital malformation of the emlbryo. Accoroding to the seasonal epidemic characteristics of RuV, women in pregnancy had better not to go to the public places during the sping seasons (March, April, May) as far as possible. PMID- 12569782 TI - [Tolerance of HepG2 and HepG2.2.15 cell lines to different apoptotic stimuli]. AB - OBJECTIVE: To elucidate the effect of HBV on hepatocyte apoptosis. METHODS: The HepG2 cells and the HBV transfected HepG2.2.15 cells were cultured with MIX or ActD or cultured in DMEM medium without serum, The apoptosis was examined with FCM. RESULTS: The apoptotic rates of HepG2.2.15 cells at 24 h and 48 h after MTX addition were 10.8% and 13.3%, at 24 h and 48 h after Act D addition were 16.8% and 37.7%, and at 4th and 6th day after serum withdrawal were 13.2% and 14.8%, respectively. While those of HepG2 cells were 12.6% and 65.3%, 44.5% and 89.7%, and 19.8% and 28.8%, correspondingly. CONCLUSION: HepG2.2.15 cell was tolerant to these apoptotic stimuli, and it might be inferred that HBV inhibits hepatocyte apoptosis. PMID- 12569783 TI - [Cloning and sequence analysis of 55 K protein of egg drop syndrome virus]. AB - OBJECTIVE: For understanding the characteristics of genomic structure of egg drop syndrome virus(EDSV). METHODS: Nucleic acid was extracted using routine method from weak virulent strain AA-2 of EDSV isolated from Chinese sick hens. Construction of the whole genomic library was by hydrolysis with Hind III, strand encoding 55 K gene locating in Hind III--A segment was sequenced and analyzed. RESULTS: The open reading frame has a length of 1,014 nt and codes a polypeptide of 337 amino acids with molecular weight of 38,200. Analysis of the amino acid sequence revealed a homology from 25.5%-32.4% to the 55 K protein of human adenovirus types 2, 12, 40, canine adenovirus and fowl adenoviruses of group 1, whereas to ovine adenovirus is 46.4%. CONCLUSION: The genomic structure of EDSV has some relationship with adenoviruses. PMID- 12569784 TI - [Study on HBV preS/S gene mutation in peripheral blood mononuclear cells]. AB - OBJECTIVE: To study the significance of mutations of HBV preS and S genes in peripheral blood mononuclear cells (PBMC). METHODS: Srea and PBMCs of 19 chronic hepatitis B patients were collected. The S, preS1 and preS2 gene fragments of HBV were separately amplified by PCR, and 5 cases of preS1 and preS2 gene fragments in paired sera and PBMC were sequenced. RESULTS: The positive rates of S, preS1 and preS2 gene fragments in serum and PBMC were 100%, 94.7%, 100% and 0, 26.3%, 26.3% respectively. Sequence analysis of preS1 and preS2 genes showed that in 8/10 specimens there were point mutations in preS1 and 7/10 mutations in preS2; while in the others (2 of preS1 and 3 of preS2) the sequences were identical to that of wild-type HBV. CONCLUSION: No full-length HBV genome in PBMC was detected by PCR, this is a fact not supporting that HBV can replicate, assemble and release from PBMC. The amino acid changes predicted mutatieds from nucleotide were focused in aa21-47 region, which might affect adsorption and penetration of HBV, and alteration in tissue tropism. Whether the presence of HBV subgenomic fragments in PBMC can affect cell function should be pursued. PMID- 12569786 TI - [Identification and characterization of HIV-1 subtype D strain in China]. AB - OBJECTIVE: Through sequence analysis of HIV-1 gag, tat and env regions, the subtype D HIV-1 had been identified appering in China. METHODS: The blood was collected from HIV-1 infected individual (SC9 712) in Sichuan Province. The peripheral blod mononuclear cells (PBMCs) were isolated from the blood by ficoll and the genomic DNA was extracted. The gag gene, the C2-V3 of the env gene and the first exon of tat gene were amplified by nested-PCR and sequenced directly. RESULTS: The genetic distance respectively is 7.56% in tat gene between SC9 712 and UG274A, 9.59% and 9.94% in gag gene between SC9 712 and K31 and VI203, 13.86% in env gene between SC9 712 and MAL. On the phylogenetic tree the SC9 712 HIV-1 strain clustered with the Africa subtype D strains UG274A, K31, VI203 and MAL. The above data demonstrated that SC9 712 was subtype D HIV-1 strain. Epidemiological data showed that SC9 712 virus was from a person who had worked in Africa. CONCLUSION: The results indicated that the person who came back from Africa had transmitted the subtype D of HIV-1 from Africa to China. PMID- 12569785 TI - [Study of the 3'noncoding region of Chinese hepatitis C virus genome]. AB - OBJECTIVE: To analyze the 3' noncoding region (3' NCR) of HCV genome from Chinese hepatitis C patients so as to facilitate further study of mechanism of HCV gene replication. METHODS: Two different strategies were employed to amplify the full length of the 3' noncoding region of HCV genome from sera of HCV infected patients in Shanghai area: one was to amplify the full-length fragment directly by nested PCR and the other amplify two overlapping fragments. The PCR products were further analyzed by sequencing and nucleotide alignments. A HCV genome 3'NCR based RT-PCR was developed and its specificity and sensitivity for HCV RNA detection in sera was compared with the established 5'NCR based RT-PCR. RESULTS: Sequence analysis showed that Chinese HCV genomic 3' NCR consists of three parts: the 5' region, poly (U-UC) tract and the 98-base region. Sequence alignments revealed that, while the 98-base regions were completely conserved in different isolates and were identical to the reported sequences, the poly (U-UC) region shared highly diversities. A high degree of concordance(95%) between the 3'NCR and 5'NCR RT-PCR for detection of HCV RNA in sera was found. CONCLUSION: The high conservation at the 3' NCR(98 bases) of HCV genome among different isolates indicated that this region may be critical for HCV gene replication The 3'NCR based RT-PCR may be a useful addition to available systems to diagnosis HCV infection. PMID- 12569787 TI - [Study on transient infection of T cell lines by M tropic HIV-1 strains]. AB - OBJECTIVE: To reveal the mechanism of transient infection of T cell by HIV-1 isolates of early stages from Yunnan and Xinjiang, China. METHODS: We made these viruses pass on CXCR4 expressing T cell lines and CCR5 expressing U937 cell line. After having observed the biological phenotype, we analyzed sequences of env gene to find genetic mutations of the strains, and used heteroduplex mobility assay (HMA) to show the complexity of the virus groups. RESULTS: Sequence analysis indicated that these viruses are M tropic, NSI strains, correlating with their phenotype; they mutated largely through T cell passage, all showed sequence characteristics deviating from M tropic/NSI to different extents. HMA results indicated the complexity of virus groups was low at the very beginning of the passage and kept high later. CONCLUSION: We concluded that these viruses tried mutating to different directions to adapt the T cell line but all failed. Although env gene correlates with cell tropism, coreceptor usage and HIV syncytium inducing, it may have relationship with the whole genome, whether these phenotypes are present or not. The results imply that there is no T tropic/SI strain in the M topic/NSI virus pool in early infection, it is generated later following continued infection in vivo. PMID- 12569788 TI - [Construction and characterization of non-replicating recombinant vaccinia virus expressing rabies glycoprotein]. AB - OBJECTIVE: To improve the safety of recombinant vaccinia rabies virus, the non replicating recombinant vaccinia rabies virus VTKRG delta CK was constructed. METHODS: The rabies glycoprotein(RG) gene was introduced into the TK locus of Chinese Tiantan strain of vaccinia virus (VTKRG), and a fragment between fragment C and fragment K involving vaccinia virus virulence and host-range related genes were deleted by homologous recombinant. RESULTS: The ability to replicate in primary chick chick embryo fibroblast(CEF) and dramatically reduced replicating capability in mammalian cells of the virus were observed, whereas VTKRG delta CK could express high level of RG efficiently. The expression of RG could be detected in CEF by indirect immunofluorescence assay and Western blot analysis. The VTKRG delta CK could elicit neutralizing antibody against CVS rabies virus and protect the animals from lethal rabies virus challenge in inoculated mice. CONCLUSION: VTKRG delta CK possesses immunogenicity and higher safety. PMID- 12569789 TI - [The experimental and clinical study of inhibitory effects of Re-Du-Qing on human cytomegalovirus (HCMV)]. AB - OBJECTIVE: To study the theurapeutic effect of Chinese medicine Re-Du-Qing on HCMV. METHODS: The expression of the HCMV late-mRNA in infected cells was measured by quantitative RT-PCR and the development of cytopathic effect (CPE) caused by HCMV was observed before and after the treatment of Chinese medicine "Re-Du-Qing". On the basis of its anti-HCMV activity study in vitro, Re-Du-Qing was applied to the child-bearing aged women and to pregnant women who were HCMV IgM positive which indicated the active HCMV infection, meanwhile the serum levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 (IL-2) were measured before and after the treatment by the ELISA method. RESULTS: The results showed that the Re-Du-Qing had strong inhibition effect both on HCMV-mRNA and CPE. The serum level of the TNF-alpha increased and IL-2 decreased significantly in HCMV-IgM positive cases when compared with normal women. After being treated with Re-Du-Qing, the serum TNF-alpha and IL-2 in patients returned to normal level, and 72.9% of the HCMV-IgM positive cases turned negative which was significantly higher than that (13.6%) in control group. CONCLUSION: It was suggested that serum TNF-alpha level is closely related to the activity of HCMV which has harmful effect on immune system. Re-Du-Qing can enhance immunity of the body and has an obvious inhibition on HCMV in Vitro and Vivo, and can thus serve as safe and effective medicine for treatment of active HCMV infection, especially for the pregnant women. PMID- 12569790 TI - [Comparison of three methods in detection of rotavirus infection in neonates]. AB - OBJECTIVE: To compare, three methods in detection of rotavirus infection in neonates. METHODS: 252 fecal samples from neonates with acute gastroenteritis, healthy neonates and under 4-month-old babies with diarrhoea were detected with ELISA, PAGE and EM. RESULTS: Using ELISA and PAGE to check 5 fecal samples from neonates with diarrhoea, the positive number of ELISA was 4 and the PAGE was all negative. One fecal sample from neonate of one month old with diarrhoea detected with ELISA was strong positive and result with EM was negative. By checking 119 fecal samples from normal neonates of less than ten days old, the positive rate of ELISA was 54.6% and results of PAGE were all RNA negative. 128 fecal samples from less than 4-month-old babies with diarrhoea showed coincident detection rate of ELISA and PAGE (81.2%), but which was going up with increasing of their ages. We also checked 55 fecal samples from less than 4-month-old babies with diarrhoea by ELISA, PAGE, EM and the positive rates were 27.3%, 38.2% and 20% respectively, the coincident rate of the three was 63.6% and the differentiation rate 36.4%. CONCLUSION: The differentiation rate between ELISA and PAGE was obvious, as in the elisa there showed a sirong positive and in PAGE the weak. The diffesence went down with increasing of the ages of the babies with diarrhoea. This indicated that the false positive rate of ELISA existed. PAGE is more reliable and objective and is the better method to check neonate diarrhoea. PMID- 12569792 TI - [Study on etiology of acute hemorrhagic conjunctivitis in Qingdao during 1997]. AB - OBJECTIVE: To study the etiology of acute hemorrhagic conjunctivitis (AHC) in Qingdao during 1997. METHODS: During the AHC epidemic season viruses from the conjunctival swabs were isolated in Hela and Hep-2 cell cultures and were identified by neutralization test. RESULTS: All isolated virus strains were identified as (CA24) v and adenoviruses (Adv) by neutralization test with antisera of CA24 v, Echo virus 70(EV70), CA24, CA24 v of Beijing local strain, Ad3, Ad7 and Ad11. CONCLUSION: In 1997, a small epidemic of acute hemorrhagic conjunctivitis was caused by adenoviuses and CA24 v. PMID- 12569791 TI - [A study of HGV infection in liver tissue of hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the prevalence of hepatitis G virus (HGV) in hepatocellular carcinoma (HCC). METHODS: HGV NS5 antigen was detected in paraffin embedded HCC tissues from 53 autopsy patients by using immunohistochemical method with antiHGV NS5 monoclonal antibody. RESULTS: Among the 53 subsjects, 20(37.7%) had been detected of HGV antigen (HGAg) in HCC tissues. When serological markers were used, the HGAg detection rate was 2/8(25%) in non-B and non-C, 16/41(39%) in HBV and 2/2 in HCV infection groups, respectively. CONCLUSION: The infection of HGV is often seen in patients with HCC. HGV infection does not appear to account for an important independent risk factor for development of HCC. PMID- 12569793 TI - [Observation on effect of chongcao shenkang capsules in preventing and treating acute renal failure in cases of hemorrhagic fever with renal syndrome]. AB - OBJECTIVE: To observe the effect of Chongcao shenkang capsules in preventing and treating acute renal failure in cases of hemorrhagic fever with renal syndrome (HFRS). METHODS: 150 cases of HFRS were randomly divided into two groups: the treatment group and the control group. The former contained 76 cases and the latter the 74 cases. All cases in both groups were given a combined therapy mainly composed of balanced salt solution and ribavirin, cases in treatment group additionally accepted Chongcao shenkang capsules 2.7 grams tid, till the end of polyuric phase. RESULTS: The results showed that in the treatment group the occurrence rate of cases with oliguric phase was 13.0% (4/46) and the oliguric and polyuric periods were 2.7 +/- 1.6 days and 6.8 +/- 2.9 days respectively while those in the control group were 31.1% (4/45), 4.3 +/- 2.1 dnys and 8.2 +/- 3.4 days respectively (P < 0.01 or P < 0.05). The urinary protein of the treatment group disappeared within 5.1 +/- 2.3 days and their serum creatinine restored within 10.9 +/- 6.2 days, but the urinary protein disappearing and the serum creatinine restoring periods of the control group were 6.8 +/- 2.7 days and 13.7 +/- 7.4 days respectively (P < 0.01 or P < 0.05). The serum and urine beta 2 microglobulin levels of the treatment group were remarkably lower than those of the control group on the fifth and tenth days after treatment (P < 0.01 or P < 0.05). Compared with those of the control group of 33.8% (25/74) and 87.8% (65/74), the occurrence rate of severe complications and curative rate of the treatment group were 18.4% (14/76) and 94.7% (72/76) respectively (the former P < 0.05, but the latter P > 0.05). CONCLUSION: It was concluded that Chongcao shenkang capsules is capable of reducing the occurrence rate of acute renal failure in HFRS patients, shortening the duration of acute renal failure, decreasing their needs for dialysis, lessening their complications and increasing their curative rate. Its administration is safe and convenient and therefore it is worthy to be popularized. PMID- 12569794 TI - [Study on a purified and inactivated Japanese encephalitis vaccine prepared on Vero cells using SA14-14-2 attenuated virus strain]. AB - OBJECTIVE: To develop a kind of new Japanese Encephalitis (JE) vaccine prepared on Vero cells. METHODS: A JE attenuated virus strain SA14-14-2 was adapted on Vero cells for preparation of purified inactivated vaccine. Comparison of the growth curves of SA14-14-2 in roller bottle and in spinner flask was made. After the virus inoculation and absorption on Vero cells for 2 hours, cultures were replaced with serum-free MEM, the culture supernatants were harvested on day 2,4,6 after inoculation. The virus fluids were pooled, concentrated by 8% PEG, and purified on 15%-60% sucrose density gradients. The purified virus was inactivated with 0.02% formalin. RESULTS: It showed that virus titer was higher and maintained longer in roller bottle. Mice vaccinated twicely with 0.5 microgram dose of the purified inactivated vaccine induced neutralizing antibody titers equal to that of the mice vaccinated with primary hamster kidney inactivated vaccine. CONCLUSION: This Vero cell prepared purified and inactivated JE vaccine made by SA14-14-2 strain could be used for human as a kind of new JE vaccine. PMID- 12569795 TI - [Analysis of TTV infection in patients of hemodialysis and bone marrow transplantation]. AB - OBJECTIVE: To make clear the infection of TT virus(TTV) in patients of hemodialysis and bone marrow-transplantation and to evaluate the virulence. METHODS: The TTV infection were investigated in 37 cases of hemodialysis and 90 cases of bone marrow-transplantation by means of review method. RESULTS: The positive rate of TTV was 16.22% (6/37) in 37 cases of hemodialysis and 3.33% (3/90) in 90 cases of bone marrow-transplantation. The difference between them was significant(P < 0.01). Of the six TTV-positive patients, 5 had blood transfusion and 4 of them had bone marrow-transplantation. Infection of TTV had nothing to do with the infection of HBV and HCV. There were no significant differences in the results of liver function tests between TTV positive and TTV negative patients. CONCLUSION: These results suggest that blood transfusion might be one of the critical routes to transmit TTV. The virulence of TTV might be mild. Whether TTV is a hepatitis viral remains further study. PMID- 12569796 TI - [Advance in the study of hepatitis B virus gene expression and replication in animal cultured cells and its uses]. PMID- 12569797 TI - [Preliminary study of cryopreservation of tissue engineered tendon]. AB - OBJECTIVE: To investigate a cryophylactic agent (CPA) to protect tissue engineered tendon (TET) in deep low temperature. METHODS: Sixty-four BALB/C inbred nude mice were chosen, which included 4 as blank control group, left sides of 60 as experimental group and their right sides as control group. Transformed human embryonic tendon cells of the 54th passage and artificial materials of carbon fiber (CF) and polyglycolic acid (PGA) were co-cultured in vitro to construct TET. TET was frozen in liquid nitrogen with four kinds of CPA (groups A, B, C, and D) for 2 months. They were thawed quickly and transplanted into hind limbs of nude mice to repair the defects of Achilles tendon, which was 5 mm in length and 65.7% of total Achilles tendon. In control group, no cryopreservation treatment was taken. The morphological, histological, ultrastructure, and immunohistochemistry examinations were made and short tandem repeat loci were detected 2, 4, 6, 8, and 12 weeks later. RESULTS: In the experimental group, the morphological properties of tendon cells resumed gradually and the capability of synthesizing collagen enhanced by degrees. Tendon cells survived and could secret type I collagen and there was less difference between experimental and control groups 12 weeks after transplantation. In group A, vacuole in mitochondrion of tendon cell decreased, tendon cell arranged in order and abundant collagen fibers were found and linked. CONCLUSION: The cryopreservation agent in group A can protect TET in deep low temperature. PMID- 12569798 TI - [Experiment of histocompatibility and degradation in vivo of artificial material calcium polyphosphate fiber]. AB - OBJECTIVE: To study the histocompatibility and degradation in vivo of a new artificial material, calcium polyphosphate fiber (CPPF), and to provide some experimental basis for further study in tendon tissue engineering. METHODS: CPPF and carbon fiber (CF) as control material were implanted in symmetry part in subcutaneous layer, muscle and Achilles tendon of 20 SD rats. The day of operation, the 4th, 8th, 12th, 16th, and 20th weeks after operation, X-ray examination was performed to detect the density change of materials. Local tissue and materials were observed grossly, and pathological examination was made with HE staining 4, 8, 12, 16 weeks after operation. RESULTS: CPPF degraded completely within 16 weeks in muscle, and in 20 weeks after implantation in Achilles tendon and subcutaneous layer according to X-ray and pathological examination. No calcium phosphate crystal deposit was observed in local tissue. No obviously degradation of CF was found within 20 weeks. Local infiltration of lymphocytes and macrophagocytes around CPPF were much fewer than that of CF. CPPF combined compactly with surrounding hyperplastic tissue. CONCLUSION: CPPF degrade thoroughly from 16 to 20 weeks without sediment of crystal of calcium phosphate in vivo. CPPF has good histocompatibility and can be used as a scaffold material of tissue engineering. PMID- 12569799 TI - [Preparation of compound biodegradable matrices and growth of vascular endothelial cell on them]. AB - OBJECTIVE: To prepare the compound biodegradable matrices, polyglycolic acid (PGA), polylactic acid (PLA) mesh and poly-beta-hydroxybutyrate(PHB) which precoated with collagen, and to observe the growth and differentiation of bovine vascular endothelial cells on these scaffolds. METHODS: By enzymatic digestion methods, bovine vascular endothelial cell (VEC) were isolated from calf thoracic aorta, then cultured and purified. PGA, PLA, PHB meshes were dipped into cross linked type I collagen solution, dried under vacuum frozen condition. VEC were seeded into these scaffolds. The growth of VEC on scaffolds was analyzed by MTT method. RESULTS: The collagen, PGA/collagen, PLA/collagen scaffolds were elasticity and tenacity. VEC grew better on collagen, PGA/collagen, and PLA/collagen membranes than on the PHB/collagen one. CONCLUSION: The PGA/collagen scaffold has elasticity, plasticity and tenacity. VEC grow best on it. It is an ideal scaffold for tissue engineered vessel reconstruction for it integrating both advantages of biomaterials and degradable materials. PMID- 12569800 TI - [Experimental study on rabbit periosteal osteoblasts and renal vascular endothelial cells indirect co-culture in vitro]. AB - OBJECTIVE: To determine an optimal co-culture ratio of the rabbit periosteal osteoblasts (RPOB) and rabbit renal vascular endothelial cells(RRVEC) without direct contact for future study of bone tissue engineering. METHODS: RPOB and RRVEC in the ratios of 1:0(control group), 2:1(group 1), 1:1(group 2) and 1:2(group 3) were co-cultured by six well plates and cell inserts. Four days later, the proliferation of RPOB and RRVEC were examined through cell count. Differentiated cell function was assessed by alkaline phosphatase (ALP) activity assay and 3H proline incorporation assay. RESULTS: When RPOB and RRVEC were indirectly co-cultured, the proliferation of RPOB and 3H proline incorporation was higher in group 1 than in the other experimental groups and control group (P < 0.05). ALP activity of RPOB was higher in group 1 than in control group and group 3 (P < 0.05), but there was no significant difference between group 1 and group 2 (P > 0.05). CONCLUSION: These results suggest that RPOB and RRVEC co cultured in a ratio of 2:1 is optimal for future study of bone tissue engineering. PMID- 12569801 TI - [Bio-derived bone transplantation with tissue engineering technique: preliminary clinical trial]. AB - OBJECTIVE: To sum up the clinical results of bio-derived bone transplantation in orthopedics with tissue engineering technique. METHODS: From January 2000 to May 2002, 52 cases with various types of bone defect were treated with tissue engineered bone, which was constructed in vitro by allogeneous osteoblasts from periosteum (1 x 10(6)/ml) with bio-derived bone scaffold following 3 to 7 days co culture. Among them, there were 7 cases of bone cyst, 22 cases of non-union or malunion of old fracture, 15 cases of fresh comminuted fracture of bone defect, 4 cases of spinal fracture and posterior route spinal fusion, 3 cases of bone implant of alveolar bone, 1 case of fusion of tarsotarsal joint. The total weight of tissue engineered bone was 349 g in all the cases, averaged 6.7 g in each case. RESULTS: All the cases were followed up after operation, averaged in 18.5 months. The wound in all the case healed by first intention, but 1 case with second intention. Bone union was completed within 3 to 4.5 months in 50 cases, but 2 cases of delayed union. Six cases were performed analysis of CD3, CD4, CD8, ICAM-1 and VCAM-1 before and after operation, and no obvious abnormities were observed. CONCLUSION: Bio-derived tissue engineered bone has good osteogenesis. No obvious rejection and other complications are observed in the clinical application. PMID- 12569802 TI - [Shape, phenotype and gap junction of the rabbit chondrocytes]. AB - OBJECTIVE: To study the gap junction and phenotype of cultured chondrocyte of rabbit, and the gap junction between the chondrocytes in the same cartilage cavities in human femoral head articular cartilage. METHODS: CFDA-AM was added into the medium of the fifth passage of chondrocyte of rabbit in the 96-well plate. The fluorescent in spherical and fibroblast-like chondrocytes was detected separately. The recurrence of the fluorescent in accordant with time in 16 minutes was recorded after blanching the fluorescent with laser. And the fluorescent after blanching of chondrocyte in the cartilage cavities in the proliferative zone of articular cartilage of adult human femoral head was recorded, too. RESULTS: The average fluorescent of the single layer of the fibroblast-like chondrocyte was 83(ranged from 1 to 274), the highest was found in the spherical shaped cell (averaged 2,057, ranged from 340 to 3,538). The recurrence of the fluorescent after the blanching appeared only in the spherical chondrocyte, the gap junctions reappeared only in the spherical chondrocytes, as well as in the cells in the cartilage cavities in the articular cartilage of the human femoral head. CONCLUSION: The appearance of the gap junction is corresponded with the spherical shape, secretion of the cartilage matrix of the chondrocyte. There are gap junctions in the cells in the same cartilage cavities in the articular cartilage of the human femoral head, while no gap junctions in the isolated chondrocytes in the cartilage. PMID- 12569803 TI - [Autocrine regulation for transforming growth factor-beta 1 by fibroblast in vitro]. AB - OBJECTIVE: To explore the autocrine regulation for the transforming growth factor beta 1(TGF-beta 1) by the fibroblasts in vitro. METHODS: Fibroblasts were cultured in vitro which isolated from the normal prepuce by circumcision. TGF beta 1 concentration were determined with ELISA for the different passages and the different time at the same passage. RESULTS: The TGF-beta 1 concentration was different for the distinct passage of fibroblasts and achieved the peak (450 ng/L) at the sixth passage. Among the same fibroblasts, the TGF-beta 1 concentration was different in the various days and passage of the summit (680 ng/L) on the fifth day, which was 2.5 times as high as that of the first day. CONCLUSION: The fibroblasts possess the ability of autocrine of the TGF-beta 1 and have some regularity. It will provide the theory basis for the research about TGF-beta 1 regulation mechanism and the medical application about salvia miltiorrhiza. PMID- 12569804 TI - [In vitro drug release behavior of carrier made of porous glass ceramics]. AB - OBJECTIVE: To conduct the in vitro test on drug release of rifampin encapsulated in a carrier made of porous phosphate glass ceramics and to analyze main factors which affect the drug release rate. METHODS: A certain quantitative of rifampin was sealed in a hollow cylindrical capsule which consisted of chopped calcium phosphate crystal fiber obtained from glass crystallization. The rifampin concentration was measured in the simulated physiological solution in which the capsule soaked. RESULTS: Rifampin could be released in a constant rate from the porous glass ceramic carrier in a long time. The release rate was dependent on the size of crystal fiber and the wall thickness of the capsule. CONCLUSION: This kind of calcium phosphate glass ceramics can be a candidate of the carrier materials used as long term drug therapy after osteotomy surgery. PMID- 12569805 TI - [Advances in research and development of tissue engineering]. AB - OBJECTIVE: From the point of view of material science, the methods of tissue repair and defect reconstruct were discussed, including mesenchymal stem cells (MSCs), growth factors, gene therapy and tissue engineered tissue. METHODS: The advances in tissue engineering technologies were introduced based on the recent literature. RESULTS: Tissue engineering should solve the design and preparation of molecular scaffold, tissue vascularization and dynamic culture of cell on the scaffolds in vitro. CONCLUSION: Biomaterials play an important role in the tissue engineering. They can be used as the matrices of MSCs, the delivery carrier of growth factor, the culture scaffold of cell in bioreactors and delivery carrier of gene encoding growth factors. PMID- 12569806 TI - [Pluripotential of human adult stem cells and its application in reparative and reconstructive surgery]. AB - OBJECTIVE: To explore the pluripotential and possible clinical application of adult stem cells. METHODS: The original articles on adult stem cells were extensively reviewed and the recent advances were summed up. RESULTS: Adult stem cells were located at different tissues of human beings and had the pluripotentiality of self-renewal and differentiation. Some adult stem cells, such as in marrow, nerve, muscle, fat, skin, liver, tissues, had the ability to differentiate into the unrelated cell type. CONCLUSION: The pluripotential, ubiquitous distribution and plasticity of adult stem cells offered a new way in regeneration medicine, such as cell therapy and tissue engineering. PMID- 12569807 TI - [Hemorheology of island flap after ischemia-reperfusion injury and modulation of dexamethasone]. AB - OBJECTIVE: To study the hemorheology of island flap after ischemia-reperfusion injury and modulation of dexamethasone. METHODS: Sixty Wister rats were made ischemia-reperfusion injury model, and divided into two groups randomly(Group I: intraperitoneal injection of normal saline 2 ml/kg as control group; Group II: intraperitoneal injection of dexamethasone 5 mg/kg as experimental group). Flap survived areas were measured and neutrophil necrosis numbers in flaps were counted. Erythrocytes and neutrophil hemorheology were observed. RESULTS: Area survived flap in group II was larger than that in group I. Neutrophil necrosis numbers were less in group II than in group I (P < 0.05). Whole blood hyposhear viscosity, erythrocyte aggregation, Casson yield stress and nerutrophil adhesion ability were higher in group I than in group II (P < 0.05); and the neutrophil deformability was lower in group I than in group II. CONCLUSION: Flap inchemia reperfusion can increase erythrocyte aggregation index and neutrophil adhesion ability. Dexamethasone can improve these and decrease neutrophil necrosis numbers, so as to prevent flap from ischemia-reperfusion injury. PMID- 12569808 TI - [Repair of circular cicatricial contracture of the thigh using deep inferior epigastric perforator flap]. AB - OBJECTIVE: To provide an ideal method for repairing the circular cicatricial contracture of thigh. METHODS: Deep inferior epigastric perforator (DIEP) island flaps was elevated based only on the deep inferior epigastric artery and vein and transferred to cover the thigh wound after scar had been resected. RESULTS: Four DIEP island flaps was applied clinically and all flaps survived. The size of the flaps ranged from (8 cm x 28 cm) to (11 cm x 32 cm). Venous return and edema had been obviously improved postoperatively. There was no abdominal weakness and hernia in the donor sites. CONCLUSION: DIEP flap not only retains the advantages of TRAM flap such as good blood supply and rich tissue volume, but also preserves the integrity of the rectus abdominis muscle. DIEP island flap is a good material for repair of the circular cicatricial contracture of the thigh. PMID- 12569809 TI - [Effect of local application of basic fibroblast growth factor and sucralfate on skin tissue structure after expansion]. AB - OBJECTIVE: To investigate the effect of local application of basic fibroblast growth factor (bFGF) and sucralfate on the skin tissue structure after expansion. METHODS: Continuous tissue expansion (CTE) experimental animal model was made with constant pressure pump in nine white pigs. The bFGF and sucralfate were perfused between tissue and tissue expander. Both bFGF and sucralfate were injected in group I, both bFGF and normal saline in group II, only sucralfate in group III, and normal saline in group IV as control group. The samples were took from each pig for histomorphological assessment on the 3rd day and 6th week after expansion. RESULTS: Compared with control group, epidermal, granular spinous and basal cell thickness increased significantly (P < 0.05) in group I; in dermal layer, dermal thickened slightly and the collagen fibers became thicker and orderly; the elastic fiber regenerated significantly; the density of fibroblasts and capillaries significantly increased (P < 0.05) in group I. The collagen fibers were broken on the 3rd day and 6th week after expansion in control group. However, the thickness of the fibrous capsule layer was similar in each group. CONCLUSION: Local application of exogenous bFGF and sucralfate combined with CTE can greatly promote the skin growth. It can accelerate the tissue expansion. PMID- 12569810 TI - [Curative effects of basic fibroblast growth factor on anus wound healing]. AB - OBJECTIVE: To observe the curative effects of basic fibroblast growth factor (bFGF) on anus wound healing. METHODS: From April 1996 to December 2000, out of 109 patients with anus trauma, hemorrhoidectomy or fistula resection, 68 were treated with bFGF as the experimental group, while 41 were treated routinely as the control group. The healing of the wound, the general and local reaction were observed. RESULTS: The healing time of the experimental group was(17.00 +/- 1.54) days while that of the control group was(20.00 +/- 1.16) days (P < 0.01). Three weeks after operation, the healing rates of the experimental and control groups were 97.1% and 87.8%, respectively (P < 0.01). No general or local detrimental reactions were found in two groups. CONCLUSION: Local application of bFGF can accelerate the healing of anus wound, and the patients have little pain. PMID- 12569811 TI - [Application of decalcified dental matrix in bone reparation]. AB - OBJECTIVE: To investigate the effect of decalicified dental matrix (DDM) on healing of traumatic fracture and its side-effect. METHODS: From June 1997 to December 1998, 42 patients with closed traumatic long bone fracture (36 males and 6 females, aged from 18 to 57 years with an average of 32 years) were divided into two groups randomly; open reduction and internal fixation were carried out in all patients, but the DDM was used only in experimental group. After operation, the body temperature, ALT, and bone union time were observed and recorded. RESULTS: Following-up 1 to 3 years, the bone union time of experimental group was significantly shorter than that of control group (P < 0.01), there was no significant difference in the temperature and ALT changes between two groups(P > 0.05). No infection occurred. CONCLUSION: DDM can promote the growth of bony callus and enhance the healing of fracture. There is no side-effect. PMID- 12569812 TI - [Clinical analyses of cervical fracture-dislocation without spinal cord injury]. AB - OBJECTIVE: To investigate the etiology, pathological mechanism and treatment of cervical fracture-dislocation without spinal cord injury. METHODS: Nine patients with cervical fracture-dislocation without spinal cord injury were male and aged 22 to 63 years. Based on the clinical symptoms and roentgenographic changes, the injury mechanism was analyzed; and the pathological characteristics and treatment principle were put forward. RESULTS: Anterior reduction was employed in all 9 cases. Eight cases were reduced completely while 1 case was reduced partially. After following up 1 to 3 years, 7 cases recovered completely and the other 2 cases relieved their symptoms obviously. No nervous symptoms aggravated during the following-up period. CONCLUSION: Fracture-dislocation of the cervical spine without spinal cord injury has special pathological mechanism. The surgical intervention is needed for solid fixation and complete decompression without any delayed neurosymptoms. PMID- 12569813 TI - [Biological characteristics of mesenchymal stem cells in vitro derived from bone marrow of banna minipig inbred line]. AB - OBJECTIVE: To isolate and characterize mesenchymal stem cells (MSCs) derived from bone marrow of Banna minipig inbred line (BMI). METHODS: BMI-MSCs was isolated from bone marrow by density gradient centrifugation and cultured in DMEM (containing 15% bovine serum) at 37 degrees C with humidified 5% CO2. These cultured stem cells were characterized in clonal growth, expression of specific markers and capability of differentiation. RESULTS: Mesenchymal stem cells were proliferative and could be expanded rapidly in vitro. Clonal growth of these cells can be observed when small amount of cells was inoculated. These cells were SH2, SH3, SH4, SB10 and SB21 positive. And it was proved that these cells possess osteo-differentiation ability, up-regulated alkaline phosphatase expression and calcium secretion after osteosupplement was added into the media for several days. CONCLUSION: Mesenchymal stem cells derived from bone marrow of BMI possess the general characters of stem cell. PMID- 12569814 TI - [Expression and distribution of xenoantigen alpha-Gal in intervertebral disk of Chinese banna minipig inbred line]. AB - OBJECTIVE: To investigate the expression and distribution of xenoantigen in intervertebral disk of Chinese banna minipig inbred line, and to study the availability of xenograft transplantation of intervertebral disk. METHODS: Samples of intervertebral disk were collected from six Banna pigs of 8 to 11 month-old. The fixation, embedment and slice were performed. alpha-Gal specific binding lection (BSI-B4) were used as affinity reagents and affinity immunohistochemistry assays (SABC methods and DAB stain) were conducted to detect the expression and distribution of xenoantigen (alpha-Gal). RESULTS: alpha-Gal was found in chondrocyte cell and chondrocyte-like cell in intervertebral disk which have the positive yellow-stained particulate aggradation. There was no stain in the matrix, elastic fiber and collagen fiber. CONCLUSIONS: The distribution of xenoantigen is locally in the tissue of intervertebral disk and its expression is weak. This suggests that the intervertebral disk of Banna pig may be alternative donor for xenotransplantation. PMID- 12569815 TI - [Study on the relationship between provenance, leaf type and quality in Magnolia officinalis]. AB - An analysis of effective ingredients in 20 to 29 year old Magnolia officinalis of three different leaf types coming from various sources shows that the content of phenols differ extremely significantly in various leaf types which are a key to determination of quality of Magnolia officinalis also varies significantly among different sources which only have a significant effect on the content of magnolol. Varieties and types cultivated in different places are responsible for the variation in content of magnolol. The conclusions obtained from this study coincide with the traditional insight into quality of Magnolia officinalis. PMID- 12569816 TI - [Prospect on the callus culture of Dentrobium Candidum used as drugs]. AB - This paper reports a comparison between the callus culture of Dentrobium Candidum and its original plant from polysaccharide (one of active constituents). The result suggests that the callus culture may be used instead of its original plant. PMID- 12569817 TI - [Comparing Swerita daridi with Swertia mileencis on pharmacologic action]. AB - The article compared Swertia daridi with Swertia mileencis about chemical composition and pharmacological action. The study showed two species drug's bitter taste chemical composition and pharmacological action are approximately equal. It can be tried to replace Swertia daridi with Swertia mileencis. PMID- 12569818 TI - [Study on the pharmacognosy of traditional Chinese medicine tubeimu]. AB - In this article, we have studied the traditional Chinese medicine Tubeimu on the pharmacognosy. This paper reports the thickening condition on the epidermis cell walls of the bulbuls of Tubeimu [Bolbostemma paniculatum (Maxim) Franquet] for the first time and corrects the wrong record in before literature. PMID- 12569819 TI - [The technology of processing carbonized Cirsium japonicum and quality standards of its prepared pieces]. AB - The technology of processing carbonized Cirsium japonicum was selected by using orthogonal experiment design. The result shows that the best way is to bake of 10 min at 220 degrees C. The contents of macro and trace elements in the prepared pieces significantly increased. PMID- 12569820 TI - [Steroids and lipids from Spirodela polyrrhiza]. AB - Two steroids and two lipids were isolated from the ethanolic extract of Spirodela polyrrhiza. Their structures were identified by spectral analysis and chemical evidence, which were identified as stigmasterol, monopalmitic glycerate, daucosterol and palmitic acid. Monopalmitic glycerate was discovered from this plant for the first time. PMID- 12569821 TI - [Study on the volatile oils of Murraya microphylla]. AB - The essential oils from the leaves of Murraya microphylla (Merr. et Chun) Swing. growing in Sangya city, Hainan province have been studied by GC-MS and 22 compounds have been isolated and identified. Major compounds are identified as beta-terpinene (27.86%), decahdro-1, 1,7-trimethylene-1H-cyclopop[e] azulene (24.572%), 1,2,3,4,4 alpha, 5,6,8 alpha-octahydro-7-methyl-1-(1-methyl) naphthalene (11.426%), 3-carene (10.125%), 1, 1-dimethy1-2-(3-methyl-1, 3 butadienyl) cyclopropane (5.581%). According to Mr. Bipeixi's opinion, it exists two distinct sections within Murraya microphylla (Merr. et Chun) Swing. belongs to Murraya section Bergera. Monoterpene must be the dominated volatile fractions according Li Qian's opinion, but it is a exception. Maybe it has special taxonomic significance. PMID- 12569822 TI - [Study on pharmacological actions of pingxiaozhitong pill]. AB - Pingxiaozhitong pill was administered ig. once daily for 7 successive days at dosages (4,2,2 g/kg). It was shown markelly suppressive effects on solid sarcoma 180 (S180) and Ehrlich aseites carcinoma (EAC) and synergism effect on cytoxan inhibiting S180 in mice. It had significant analgesic effect. 4,2 g/kg pingxiaozhitong pill enhanced effects on phagocysis of monocyte macrophage and value of hemagglutinin in serum in mice. PMID- 12569823 TI - [Effect of different animal skin on the transdermal speed constant of sinomenine]. AB - OBJECTIVE: To study the effect of different animal skin on the transdermal speed constant of sinomenine. METHODS: Improved franz-cell was used for the transdermal study, pH6.8 phosphate buffer was used as the receptor and HPLC method was used for the determination of sinomenine through different animal skin. RESULTS: The transdermal speed constant of the skin of hareless mice was much close to which of person. The higuchi equation of sinomenine through skin of hareless mice stored under -20 degrees C for 0d, 3d and 7d were Q = 40.34t1/2 + 196, Q = 38.01t1/2 + 25.3, Q = 41.55t1/2 + 24.7 respectively. CONCLUSION: The results indicate that skin of hareless mice is suitable for the transdermal study of sinomenine, and the effect of storing time (0-7d) under -40 degrees C is not obvious. PMID- 12569824 TI - [The habit of flowering and fruiting of Scaphium lychnophorum]. AB - The flowering and fruiting are much different between the trees of Scaphium lychnophorum which were introduced in Jinghong. The trees have on-year of fruiting once 3-4 years and off-year continue 2-3 years. In Jinghong, the low temperature in the early flowering period is one factor among the main factors effecting flowering and fruiting of trees. The trees which vegetative grow flush and which grow on the place in a poor sunlight are effected their flowering and fruiting, too. PMID- 12569825 TI - [Identification of Chinese drug sea coconut]. AB - Sea Coconut is the seed of Lodoicea maldivica Persoon, palmae. It is one species in the Seychelles Islands, Africa. The drug is yellow-white in colour, slice shaped, 12 cm in length and 4 cm in breadth. Seed-coat cell circular in shape. Under seed-coat, the cells are elliptic, and stone cells are rectangle with thick wall and large pit. Endosperm cell elongate with thick wall and large pits, the end of pits enlarge as a pearl. PMID- 12569826 TI - [Microscopic identification of Herba Dianthi grown in Shandong]. AB - The medicinal materials of four species and two varieties of Herba Dianthi grown in Shandong were identified. The result shows that they are identified easily and accurately according to the outer properties, and they are apparently divided into the Shizhu group, the Qumai group and the Dianthus shandongensis on the basis of their morphological and structural characteristics of the stem and leaf, but they have not obvious distinction among the species of every group. PMID- 12569827 TI - [Microscopic and TLC identification on the fruits of ten species plants for Umbelliferae]. AB - The fruits of ten species plants of Umbelliferae, including the fruits of Peucedanum decursiyum, Saposhnikovia divaricata, Peucedanum terebinthaceun, Anethum graveolens, Cnidium monnieri, Angelica sinensis, Foeniculum vulgate, Angelica polymorpha, Ferula tunnshanica and Cicuta virosa were identified on histology and TLC. PMID- 12569828 TI - [Investigation of injurious insects in storehouses of medicinal materials in Xinjiang]. AB - By investigation it was discovered that the injurious insects in storehouses of medicinal materials in Xinjiang had 95 species belonging to 29 families of 7 orders, 32 species of them were main. The register, distribution and injurious kinds of medicinal materials were listed. PMID- 12569829 TI - [Isolation and identification of triterpenoide compound from Patrinia scabiosaefolia]. AB - In this paper, a triterpenoide was isolated from the roots and rhizomes of Patrinia scabiosaefolia Fish ex Link. Its structure was identified by combination of chemical reactions and spectrum analysis as 28-O-beta-D-glucopyranosyl-(1-->6) beta-D-glucopyranosyl-hederagenin ester. The compound was found in the Patrinia for the first time. PMID- 12569830 TI - [Isolation and identification of active constituent of Toddalia asiatica in cardiovascular system]. AB - The ethereal extract from the crude ethanolic extract of Toddalia asiatica was chromatographed on silica gel and tested on biological activity. A crystalline was obtained and confirmed as an active constituent in cardiovascular system. The active constituent was identified as isopimpinellin on the basis of chemical method and spectral analysis. PMID- 12569832 TI - [Study on the Rheum palmatum volatile oil against HBV in cell culture in vitro]. AB - The antiHBV effect of Rheum palmatum Volatile oil was studied by using 2215 cell line transfected with HBV DNA. At the same time MTT method was applied for the detection of cytoxicity of drugs, selecting acyclovir(ACV) as control medicine. It turns out that the toxic concentration of Rheum palmatum Volatile oil for 50% cells was (CD50) > 1.25 x 10(-1) g/L. When concentration was below 0.625 x 10(-1) g/L, the survival rate of cells was over 90%. The maximum inhibitory rates for HBsAg and HBeAg were 70.71 +/- 5.4% and 30.99 +/- 5.3% respectively. This shows Rheum palmatum Volatile oil possesses the effect of antiHBV in vitro. PMID- 12569831 TI - [Effects of Curcuma phaeocaulis on learning and memory and lipid peroxide in mice]. AB - Nine months mice were inoculated and the method of water maze test and lipid peroxide determination to study the effects of extract of Curcuma phaeocaulis Valeton on learning and memory ability and retarding of senescence. Results showed that in water maze test the time for the group with Curcuma phaeocaulis Valeton needed from initial position to the end was significantly shortened compared with old control. The error numbers for the group of Curcuma phaeocaulis Valeton were significantly decreased compared with old control after starting experiment for 18 days and 24 days and for 35 days no significance difference was showed between groups. The levels of brain, liver and red blood cells lipid peroxide in the group with Curcuma phaeocaulis Valeton were considerably lower than old control, hemoglobin content was considerably increased than old control. PMID- 12569833 TI - [Cultivation of Pinella ternata in artificial soil and its growth habits]. AB - In order to increase yield, the cultivation of Pinellia ternata in artificial soil was studied, and its growth habits was investigated. At same time, the measures of dominating soil thickness, using nutrient solution as fertillizer and complete illumination were taken. The research results can supply a reference for the industrial production of P. ternata. PMID- 12569834 TI - [TLC identification of Flos Carthami]. AB - In 137 kinds of medicinal meterials included 21 kinds of traditional Chinese patent medicines in Pharmacopoeia such as Qili Powder, character identified constituent of Flos Carthami was found by TLC, and was successful to be used to identification of Flos Carthami in 9 kinds of traditional Chinese patent meicines such as Qili Powder. PMID- 12569835 TI - [Morphological study on rhizome of Dioscorea gracillima being forgery of Rhizoma Dioscoreae Hypoglaucae]. AB - Rhizome of Dioscorea gracillima is the main forgery of Rhizoma Dioscoreae Hypoglaucae. The paper reports the results of study on chracteristics of its apperance, micriscopical characteristics of cross section and powder, characteristics of inner wall of trachea by scanning electron microscope. Rhizome of Dioscorea gracillima can be distinguished from Rhizoma Dioscoreae Hypoglaucae by above results of study. PMID- 12569836 TI - [The history of Sparganium stoloniferum processing]. AB - Through the study on medical literatures about Sparganium stoloniferum processing from Tang Dynasty to Qing Dynasty and Pharmacopoeias of China and the standards of herbal processing practised in defferent localities in China, the author summed the history of Sparganium stoloniferum processing, and its original meaning and gave suggestions to improve the Sparganium stoloniferum processing. PMID- 12569837 TI - [Isolation and identification of alkaloids form Menispermum dauricum growing in Xianning]. AB - The alkaloids of rhizoma of Menispermum dauricum DC growing in Xianning have been subjected to isolation and identification. The results showed that its two major constituents, which are only next of dauricine in content, are dauricinoline and daurinoline, instead of the commonly found daurisoline in the same plant materials from North China. PMID- 12569838 TI - [Study on chemical components of Momordica charantia]. AB - The paper deals with the study of chemical constituents of the unmatured fruits of Chinese traditional medicine Momordica charantia L. which is usually used as green-stuff. There are two parts of the extracts obtained by ethanol precipitation, and four compounds obtained from the further isolation. They are identified as Vincine, Mycose, Momordicoside A and Momordicoside B. PMID- 12569840 TI - [Effects of polysaccharide of Polygala aureocauda on the immunnity functions of normal mouse]. AB - The effects of polysaccharide of polygala aureocauda on the immunity functions of the normal mouse were researched by the tests of clearing away carbon granules, hemolysin antibodies production and lymphocyte transformation. The results showed that the polysaccharide of polygala aureocauda Dunn. (PADTP) could strengthen the phagocytosis of the single nucleus phagocytes, promote the production of the antibodies, and raise the transformable rate of the lympocytes. That proved the PADTP could strengthen immunity function of the normal mouse. PMID- 12569839 TI - [Effects of herba Pogostemonis on gastrointestinal tract]. AB - The effects of the three extracts (Decoction, oil-free decoction and volatile oil) of Herba Pogostemonis on gastrointestinal tract were studied. The results showed that all the three extracts inhibited the automatic contraction and Ach, BaCl2-induced spasmodic contraction of isolated rabbit intestine, among the three extracts the volatile oil was the most potent. In vivo the decoction and the oil free decoction could depress gastric evacuation and inhibit the normal and neostigmine-induced intestinal propellent movement in mice, but the volatile oil could not. The decoction and the oil-free decoction also increased gastric secretion of acid and activity of pepsin and amylase. Furthermore, the decoction and the oil-free decoction reduced the incidence of diarrhea induced by senna but volatile oil enhanced cooperatively. All the three extracts relieved the gripping pain induced by abdominal administration of acetic acid, and the effect of decoction was more potent that the others. The above results revealed that the effective components of Herba Pogostemonis may be mainly water-solube. PMID- 12569841 TI - [Effects of oncomelania-killing for the total saponins of Phytolacca acinosa]. AB - The total saponins got from the roots of Phytolacca acinosa Roxb were examined about the effect of Oncomelania-killing. The results showed the efficiency was 95.6% controlled the temperature at 28 degrees C and concentration of drugs at 125 mg/L after soaking for 24 h. PMID- 12569842 TI - [The study on the fermentation medium of Ganoderma lucidum]. AB - This study has explored the fermentation medium of Ganoderma lucidum including inorganic salts, carbon source, nitrogen source and vitamins. Then a suitable formula of fermented medium was obtained by comparing different production slats of Gandoerma powder during fermentation. PMID- 12569843 TI - [Pharmcognostial study on the Geranium wilfordii var. glandulosum]. AB - First time has done pharmcognostical research on the Geranium wilfordii Maxim var. glandulosum Z. M. Tan, including microscopic identification, chemical qualitative test and bacteriostatic test. PMID- 12569844 TI - [Morphological and histological identification of Lysimachia christinae]. AB - This paper reports the identification of morphological and histological characteristics of Lysimachia christinae. It can supply basis for the identification of the true and false medicinal materials. PMID- 12569845 TI - [Studies on the chemical components of Brucea javanica]. AB - Three active components were isolated from treated dry-fruits of Brucea javanica (L.) Merr by chromatographic methods, and they were identified as Brusatol (I), Bruceine D (II), Bruceosidae A (III) by means of UV, IR, 1H-NMR, 13C-NMR spectroscopic analysis methods. PMID- 12569846 TI - [The isolation and identification of flavonoids from Drosera burmannii]. AB - Two flavonoid compounds were isolated from the whole plants of Drosera burmannii Vahl. They were identified as quercetin (I) and hyperoside (II) by means of physicochemical properties and spectral analysis (UV, IR, 1HNMR, 13CNMR and MS). PMID- 12569847 TI - [Study on the supercritical-CO2 fluid extraction of Prunus mandshurica oils]. AB - The paper reported technological study on extracting Prunus mandshurica oils by supercritical-CO2 fluid, mainly researched the influence of pressure, temperature, time and flow rate of CO2 on the oil yield, determined optimum technology of extracting the oils, compared the oils from SFE-CO2 and traditional technology. PMID- 12569848 TI - [Effects of artemisia capillaris on blood glucose and lipid in mice]. AB - The effects of Artemisia capillaris Thunb (AC) on blood glucose and lipid in mice were studied and its effective mechanism was explained. The results showed that hyperglycemic effect in aloxan-induced mice was inhibited (P < 0.01); that hyperglycemica in DM mice was decreased (P < 0.05); but the change of fasting blood glucose (FBG) in normal and hyperlipidemia mice was not significant (P < 0.05); that the level of total cholesterol (TC) and/or atherosclerotic index (AI) was decreased (P < 0.01-0.05) and the content of high density lipoprotein cholesterol (HDL-C) was increased in the above different model mice (P < 0.01 0.05). These results indicated that the AC would have the similar hypoglycemic effect as biguanide-drugs, improve endogenous and exogenous metabolic derangement in blood lipid. These suggest that AC could be used to protect and treat DM and its chronic complication. PMID- 12569849 TI - [The pharmacologic research on "shen yang formulae"]. AB - Trough pharmacologic test on seral cholesterol and teral triglycerin in normal mouse fed by high fat food and by hperlipidemia mouse, it is found that "Sen Yang Formulae" has a very good effect on lowering cholesterol and triglycerin, and it also has good effect on promoting quantity of HDL-C, ratio of HDL-C/TC, lowering quantity of LDL-C and improving condition of blood viscosity and fluidity. The results indicate that it is effective on prevention and treatment of hyperlipidemia and benefits at prevention and treatment of arteriosclerosis and coronary heart disease. PMID- 12569850 TI - [Pharmacological studies of shundao granules on gastrointestinal track]. AB - The traditional Chinese medicine Shundao Granules po markedly inhibited the formation of gastric ulcers induced by indomethacin, restraint and pyloric ligatin in rats. A decreased volume of gastric juice collected from pyloric ligated stomaches and a reduced level of gastric acid have been observed. The results showed that Shundao Granules had markedly analgesic action and significantly enhanced intestinal movement in mice, increased propulsive rate of small ingestion in mice. PMID- 12569851 TI - [The connotation investigation of "female-male" in Chinese traditional medicine nomenclature]. AB - The connotation investigation of "female-male" in nomenclature of Polygonum multiflorum Thunb, suggests that the female-male" in Chinese medicine nomenclature is generalized concept. It has three connotation, including correlation, relativity identity. It's concrete use of materialist dialectics in Chinese medicine. PMID- 12569852 TI - [Seeds collecting of Ephedra sinica and treatment before sown]. AB - Seeds collecting of Ephedra sinica and treatment before sown were studied. These results indicated that time collecting seeds and treating such as threshing, washing, drying, storing at low temperature and immersing seeds with chemical were available measures. Which could supply a reference for the culture of E. sinica. PMID- 12569853 TI - [Ultra-morphological study on the surface of the stem and leaf of the original plant of herba Dianthi in Shandong]. AB - Ultra-morphology of 4 spieces and 2 varities of the original plant of Herba Dianthi grown in Shandong was identified with SEM. The result showed that they were aparently divided into the Shizhu group and the Qumai group, and the Dianthus shandongensis was between the two groups, on the basis of the cutin grain type of the epidermis of the stem and leaf as well as the surface of the guard cell of the stoma. PMID- 12569854 TI - [Pharmacognostical identification of Chinese drug wuxiangcao (Mosla soochowensis)]. AB - In this paper, Chinese drug Wuxiangcao (Mosla soochowensis Matsuda) is made an initial study on character, microscopical and physicochemical sides. It provide a scientific basis for its comprehensive devlopment and ultilization, and to use in clinical. PMID- 12569855 TI - [Pharmacognostical identification of Pholidota cantonensis--a confused variety of herba dendrobii]. AB - This paper reports the identification of Pholidota cantonensis on medicinal materials character, microscopical characteristic, TLC and UV. The results supply a basis for the identification of Herba Dendrobii medicinal materials. PMID- 12569856 TI - [Study on the chemical components of Rabdosia excisa]. AB - Four kinds of components were extracted and separated from the stems and leaves of Rabdosia excisa. They had been identified as Kamebakaurin, Ursolic acid, Daucosterol and Liquiritin respectively by physical and chemical methods, as well as spectrum data. It was first time that Liquiritin was obtained from the Robdosia family, and its 13C-NMR data of Liquiritin had not been reported before. PMID- 12569857 TI - [Study on the chemical components of Alpinia officinarum]. AB - Four crystalline substances were isolated from rhizome of Alpinia officinarum Hance. They were identified as beta-Sitosterol (I), Galangin (II), Emodin (III) and Quercetin (IV), I and III were separated from this plant for the first time. PMID- 12569858 TI - [The effects of jianpizengmian granule on tumor growth and immune function]. AB - The effects of Jianpizengmian (JPZM) granule on tumor growth and immune function were researched in tumor-bearing mice. The results showed it had significant inhibiting activity against S180 and HePA by oral JPZM. The inhibitory rate of JPZM reached 46.25% and 45.57%. JPZM could markedly increase the index of spleen and thymus, the level of serum hemolysin and the spleen lymphocytes transformation in tumor-bearing mice, compared to control group (H2O) respectively. PMID- 12569859 TI - [Study of old-age-resistant function of Radix Achyranthes Bidentatae]. AB - Make mice to take the decoction of Radix Achyranthes Bidentatae for 30 days. The result was that the SOD vigour in the senile model mice had been raised and the plasma LPO reduced. PMID- 12569860 TI - U.S. is lone holdout on drug access vote. PMID- 12569861 TI - State budget cuts play key role in 2003 national HIV/AIDS policies. PMID- 12569862 TI - Official wants TB, HIV tests for all U.K. immigrants. PMID- 12569863 TI - Study: few immigrants bring HIV into U.S. PMID- 12569864 TI - Fear of infection. Proof of HIV, 'channel of infection' absent in mental anguish case. PMID- 12569865 TI - State, county budget woes imperil AIDS programs. PMID- 12569866 TI - UNICEF report: HIV/AIDS an increasing threat to world's children. PMID- 12569868 TI - SFAF's injunction against activist group upheld on appeal. PMID- 12569867 TI - Appeals court upholds injunction against ACT-UP members. PMID- 12569869 TI - Malpractice claim from needlestick injury reinstated on appeal. PMID- 12569870 TI - Claims for HIV/AIDS treatment in Colombia jump more than 400 percent. PMID- 12569871 TI - FDA to fast-track VaxGen experimental AIDS vaccine. PMID- 12569872 TI - Risk assessment. Insurance firms to terminate sexual history questions. PMID- 12569873 TI - Smallpox vaccination begins in U.S.--precautions needed. AB - People with HIV and certain other medical conditions must avoid smallpox vaccination, and also vaccinia infections from others who have been vaccinated recently. PMID- 12569874 TI - U.S. blocks trade agreement on generic drug access in poor countries. AB - Governments can override patents for legitimate purposes, including public health. But many small or poor countries hve no pharmaceutical industry to manufacture medicines themselves, and global trade rules being phased in will prevent others from exporting to them without the patent holder's permission. Negotiations to solve this problem broke down when the U.S. insisted that any agreement apply only to AIDS, tuberculosis, malaria, and similar major epidemics- excluding cancer, heart disease, and hundreds of other diseases. PMID- 12569875 TI - Tenofovir (Viread(R)) access for poor countries. AB - Gilead Sciences announced that it will make its drug tenofovir available at cost in all of Africa, and in some of the poorest countries elsewhere. PMID- 12569876 TI - New government AIDS Web site, phone number. AB - A new site, www.aidsinfo.nih.gov, has government-approved treatment information, as well as searchable listings of government and private clinical trials. PMID- 12569877 TI - The Center for AIDS and Humanity presents an exhibit by artist Robert Sherer. PMID- 12569878 TI - Are you willing to fight for your healthcare? PMID- 12569879 TI - "Dental care: all I want for the holidays is to keep my two front teeth". PMID- 12569880 TI - Faith-based HIV services. PMID- 12569881 TI - Segmental BIA: can we measure lipodystrophy? Emerging data reported at the Ninth Annual ANSA Conference. PMID- 12569882 TI - The positive outcomes of HIV palliative care consultations: five meaningful cases. PMID- 12569883 TI - How common illicit substances interact with antiretroviral agents. PMID- 12569884 TI - HIV in pregnancy: an update of current recommendations. PMID- 12569885 TI - HIV infected are more at risk for intimate partner violence. PMID- 12569886 TI - Evidence-based dental practice and HIV: is there relevance? PMID- 12569887 TI - Expertise is needed to provide medical nutrition therapy in HIV. PMID- 12569888 TI - HIV and substance use: pain and the drug-seeking patient. PMID- 12569889 TI - Simplifying HIV therapeutics, and the global treatment of AIDS. PMID- 12569890 TI - A solitary brain lesion in a patient with AIDS. PMID- 12569891 TI - Double "d" drug danger. PMID- 12569892 TI - Update on antiretroviral drug resistance testing: combining laboratory technology with patient care. AB - HIV drug resistance testing has been considered an emerging asset to modernized HIV management. Despite the increased number of antiretroviral agents currently available, virologic failure remains a significant problem. Drug resistance testing is designed to identify gene mutations or viral growth characteristics that suggest reduced drug susceptibility. The widespread use and virologic benefits of resistance testing in some prospective clinical trials have prompted the development of formal guidelines by expert panels for clinical use. Despite technological advances in drug resistance testing, clarification of assay interpretation, assay standardization, and the results from validation studies are needed. This review discusses updated genotyping and phenotyping methodologies, assay utilities and limitations, clinical validation studies, and current recommendations. PMID- 12569893 TI - Editorial comment: resistance testing--an integral part of HIV management. PMID- 12569894 TI - Virologic testing in the management of perinatal HIV exposure. AB - Over the last several years, virologic assays for the detection and measurement of HIV-1 RNA have become important in the diagnosis and management of perinatal HIV infection. Since AIDS Clinical Trials Group 076, a number of prospective and retrospective analyses have investigated the role of viral load in perinatal transmission. Although no universal virologic threshold for perinatal HIV transmission has been established, much has been learned about the timing of perinatal transmission and the relationship between maternal viral load and disease progression in HIV-infected neonates. Virologic assays have become accepted as standards of care in monitoring viral load during pregnancy, diagnosing neonatal infection, and establishing prognosis and response to therapy in infected infants. PMID- 12569895 TI - WHO/CONRAD technical consultation on nonoxynol-9, World Health Organization, Geneva, 9-10 October 2001: summary report. AB - This report summarises the evidence presented to a technical consultation on the safety of N-9 and its effectiveness for protection against pregnancy, sexually transmitted infections and HIV, organized by the World Health Organization Department of Reproductive Health and Research with CONRAD in October 2001. Key conclusions from the meeting include: 1. Although Nonoxynol-9 has been shown to increase the risk of HIV infection when used frequently by women at high risk of infection, it remains a contraceptive option for women at low risk. 2. Nonoxynol 9 offers no protection against sexually transmitted infections such as gonorrhoea or chlamydia. 3. There is no evidence that condoms lubricated with nonoxynol-9 are any more effective in preventing pregnancy or infection than condoms lubricated with silicone, and such condoms should no longer be promoted. However, it is better to use a nonoxynol-9 lubricated condom than no condom at all. 4. Nonoxynol-9 should not be used rectally. PMID- 12569896 TI - [Medicine as a profession has four feasible obligations]. PMID- 12569897 TI - Optical and structural properties of Ag-Si3N4 nanocermets prepared by means of ion-beam sputtering in alternate and codeposition modes. AB - Ion-beam sputtering deposition has been used in two ways, as granular multilayers and as cosputtered film, to elaborate Ag-Si3N4 nanocermets. Multilayer deposition creates slightly oblate clusters, and cosputtering produces two cluster families: elongated clusters within the Si3N4 matrix and larger ones at the film surface. The transmittance spectra of these nanocermets are characterized by a surface plasmon resonance. In the reported research the position of this resonance is related to the morphological properties of silver nanoclusters, which are studied by transmission-electron microscopy, grazing-incidence small-angle x-ray scattering, and atomic-force microscopy. PMID- 12569899 TI - 80% of all cases of blindness can be prevented or treated. PMID- 12569898 TI - Characteristics of effective clinical guidelines for general practice. AB - BACKGROUND: The use of clinical guidelines in general practice is often limited. Research on barriers to guideline adherence usually focuses on attitudinal factors. Factors linked to the guideline itself are much less studied. AIM: To identify characteristics of effective clinical guidelines for general practice, and to explore whether these differ between therapeutic and diagnostic recommendations. DESIGN OF STUDY: Analysis of performance data from an audit study of 200 general practitioners (GPs) in The Netherlands conducted in 1997. SETTING: Panel of 12 GPs in The Netherlands who were familiar with guideline methodology. METHOD: A set of 12 attributes, including six potential facilitators and six potential barriers to guideline use, was formulated. The panel assessed the presence of these attributes in 96 guideline recommendations formulated by the Dutch College of General Practitioners. The attributes of recommendations with high compliance rates (70% to 100%) were compared with those with low compliance rates (0% to 60%). RESULTS: Recommendations with high compliance rates were to a lesser extent those requiring new skills (7% compared with 22% in recommendations with low compliance rates), were less often part of a complex decision tree (12% versus 25%), were more compatible with existing norms and values in practice (87% versus 76%), and more often supported with evidence (47% versus 31%). For diagnostic recommendations, the ease of applying them and the potential (negative) reactions of patients were more relevant than for therapeutic recommendations. CONCLUSION: To bridge the gap between research and practice, the evidence as well as the applicability should be considered when formulating recommendations. If the recommendations are not compatible with existing norms and values, not easy to follow or require new knowledge and skills, appropriate implementation strategies should be designed to ensure change in daily practice. PMID- 12569900 TI - Need to boost funding for TB control in worst affected areas. PMID- 12569901 TI - New WHO publication explores important links between health and human rights, an area drawing increased attention. PMID- 12569902 TI - Insomnia sufferers failing to seek medical help in Europe and USA. PMID- 12569904 TI - Current readings in nuclear medicine. PMID- 12569903 TI - Violence and health. PMID- 12569905 TI - Probiotics in prevention of antibiotic-associated diarrhea: meta-analysis. PMID- 12569907 TI - Death in children with epilepsy: a population-based study. PMID- 12569906 TI - Risperidone in children with autism and serious behavioral problems. PMID- 12569909 TI - Bibliography. Current world literature. Circulation and hemodynamics. PMID- 12569908 TI - Bibliography. Current world literature. Hormones, autacoids, neurotransmitters and growth factors. PMID- 12569910 TI - Bibliography. Current world literature. Hematology and oncology. PMID- 12569912 TI - Bibliography. Current world literature. Infectious diseases and immunization. PMID- 12569911 TI - Bibliography. Current world literature. Orthopedics. PMID- 12569913 TI - Conversations with John H. Glenn Jr. Interview by Frank Sietzen Jr. PMID- 12569914 TI - [Research and studies on design and production for medical diagnostic imaging systems considered environmental problems part 2]. PMID- 12569915 TI - Arrhythmogenic right ventricular dysplasia/cardiomyopathy. PMID- 12569916 TI - Major new thrust for MEMS engines. PMID- 12569918 TI - A sexual assault protocol for Catholic hospitals. PMID- 12569919 TI - Dealing with scientific disputes involving authorship. PMID- 12569920 TI - Research integrity. PMID- 12569921 TI - Scientific misconduct and research integrity: federal definitions and approaches. PMID- 12569922 TI - Keep your hands off other people's DNA. PMID- 12569923 TI - Planning end-of-life care for patients with dementia: roles of families and health professionals. AB - We examined families' end-of-life decision making and their interactions with health professionals. Twenty-eight family members of institutionalized dementia patients participated in four focus groups. We found that participating family members were not well prepared for their decision-making roles, and that they: 1) experienced substantial burdens and loss in caring for institutionalized elders; 2) had limited understanding of the natural progression of dementing conditions; 3) were uncomfortable in setting goals for their relatives' end-of-life care; 4) had little experience with death, and were ambivalent about the anticipated death of their relative; and (5) reported that they had little substantive communication with health professionals regarding end-of-life care planning. We concluded that many of the needs of such families could be addressed through improved application of the principles of advance care planning, including regular structured discussions, involvement of surrogate decision-makers, and anticipation of clinical decisions. Health professionals should take the lead in 'normalizing' the discussion of death. PMID- 12569924 TI - Older adults' attitudes toward death: links to perceptions of health and concerns about end-of-life issues. AB - The current study had two primary goals, to determine whether: 1) self-rated mental and physical health, pain, and experience with health problems were predictors of elderly adults' attitudes toward death; and 2) death attitudes predict end-of-life medical treatment concerns. Participants were 109 adults, 65 years of age or older (M=78.74 years), recruited from the local community. Regression analysis indicated that poorer perceived physical health predicted a greater likelihood of viewing death as an escape, and poorer perceived mental health predicted a greater fear of death. Viewing death as an escape and fearing death predicted end-of-life medical treatment concerns; a greater endorsement of either attitude predicted more concern. Possible explanations for the links between perceived health, attitudes toward death, and concern about end-of-life issues are suggested. PMID- 12569925 TI - End of life care and reactions to death in African-American and white family caregivers of relatives with Alzheimer's disease. AB - Family caregivers for relatives with Alzheimer's Disease (AD) often experience significant stress-related problems in mental and physical health. Patients with AD often survive for protracted periods of time, placing an extensive burden of care on the caregiver prior to the patient's death. The present study addresses ethnic differences in the experience of AD caregivers around the time of their loved one's death, including life-sustaining treatment decisions and reactions to death. The results showed that, in our sample, more patients died in their homes than has been reported for deaths in the United States. African-American and White caregivers differed substantially in their reports of end of life care and subjective reactions to the death. Compared with White caregivers, African American caregivers were less likely to make a decision to withhold treatment at the time of death, less likely to have their relative die in a nursing home, and reported less acceptance of the relative's death and greater perceived loss. Results suggest that death after AD caregiving deserves further study, and that ethnic differences in end of life care and bereavement may be of particular importance. PMID- 12569926 TI - Dynamic three-dimensional echocardiography offers advantages for specific site pacing. PMID- 12569927 TI - "Ecstasy" and neurotoxic effects on the brain. PMID- 12569928 TI - A plea for a concerted nomenclature for arbuscular myocorrhizal fungal genes. PMID- 12569929 TI - HIV/AIDS and health care workers: contact with patients and attitudes towards them. AB - Data on a range of variables associated with contact with patients with HIV/AIDS and attitudes towards them were collected from 174 Scottish health care workers. Measures of attitudes and contact (overall, social and physical) were derived from the scales devised by Pleck et al. (1988). The measure of overall contact was not significantly related to attitudes, but those with predominantly social contact with patients with HIV/AIDS had more positive attitudes towards them. This relationship was moderated by occupational characteristics, concern about working with people of unknown HIV status and neuroticism. With all these variables controlled, including social contact, those who had not received in service training relating to HIV/AIDS had more negative attitudes. The results are discussed with regard to the measurement of social contact with patients, salient beliefs, the occupational characteristics associated with attitudes, and in-service education. PMID- 12569930 TI - Human factors in accidents. 1987. PMID- 12569931 TI - Certainty and uncertainty in genetic counselling: a qualitative case study. AB - This study examines the communication that occurs during routine genetic counselling sessions. It involves the qualitative analysis of nine consultations with one member of a clinical genetics team. The consultations were characterised by a contrast between the sense of certainty apparent in how the clinicians talked about the power and promise of the new genetics and the uncertainty communicated regarding the actual information produced by genetic tests and their limitations in solving people's problems. There was also a contrast between how the clinician seemed to control the format and agenda of the consultation and the apparently uncontrollable personal and social implications of the topics discussed. We speculate that this may be explained in terms of the clinician giving some order and certainty in an area of inherent uncertainty, and where great promises are as yet unrealised. PMID- 12569932 TI - Genetic countdown: biologists take a crucial step toward synthesizing life. PMID- 12569933 TI - Biotech in the clinic. PMID- 12569934 TI - The five sexes, revisited. PMID- 12569935 TI - A structure for deoxyribose nucleic acid. 1953. PMID- 12569936 TI - Molecular structure of deoxypentose nucleic acids. 1953. PMID- 12569937 TI - Experimenting with the "right to die" in the laboratory of the states. PMID- 12569938 TI - Managed care and the health of a nation. PMID- 12569939 TI - Molecular configuration in sodium thymonucleate. 1953. PMID- 12569940 TI - Transplant Amendments Act of 1990. Public Law 101-66. PMID- 12569941 TI - [There was once a Cadet Officers School in Warsaw... (Graduates of the Medical Cadet Officers School 1922-2001)]. AB - Report on an exhibition in the building of the Special Collections Department of the Main Medical Library, devoted to the war-time and post-war fate of the graduates of the Medical Cadet Officers School, a military medical academy training physians with the rank of officer that existed in Warsaw until the outbreak of the Second World War. PMID- 12569942 TI - Survey of recent Halakhic periodical literature--surrogate motherhood. PMID- 12569943 TI - OCT findings in patients with retinopathy after watching a solar eclipse. AB - PURPOSE: To evaluate the optical coherence tomography (OCT) findings in patients with solar retinopathy after watching a solar eclipse. METHODS: Complete ocular examinations and OCT were done in 4 patients presenting with acute solar retinopathy soon after observation of an eclipse. All 4 patients repeated the examinations about 1 month and 1 year after the first visit. RESULTS: The symptoms and fundus findings were similar in all patients; all eyes were emmetropic. However, the OCT images were different in all patients, and the alterations were at different levels. The most evident alterations shown by OCT were: a reduction in the intensity of reflectiveness of the retinal pigment epithelium in 3 cases; intraretinal nonreflective spaces between the inner retinal layers in 2 cases; increased reflectiveness of the inner retinal layers in 2 cases, and a round hyperreflective formation in the vitreous just in front of the fovea in 1 case. All these OCT alterations disappeared after 1 month. CONCLUSIONS: The retinal damage arising soon after exposure to sunlight showed many different aspects in the OCT images of the 4 cases examined. All retinal layers seemed to be altered, but these alterations disappeared after 1 month, and the OCT findings remained the same after 1 year. PMID- 12569944 TI - Precedents for hospice and surrogate decision-making in Jewish law. PMID- 12569945 TI - [Inky marks of memory]. AB - An exhibition in the Special Collections Department of the Main Medical Library in Warsaw, the second in a series devoted to women's themes, presenting the correspondence of Polish women-doctors, the wives and fiansees of well-known physicians and the patients of the latter, complemented with photographic documentation. PMID- 12569946 TI - Public reason and political conflict: abortion and homosexuality. PMID- 12569947 TI - The market for surrogate motherhood contracts. AB - Surrogate motherhood is a controversial subject, and has not previously been formally modelled by economists. In this paper, a neoclassical model of the market for surrogate motherhood contracts is developed, based on the utility maximizing decisions of potential surrogate mothers and commissioning parties. The presence of both altruistic and self-interested behaviour generates unusual market outcomes. PMID- 12569948 TI - Interventional endoscopic ultrasound. AB - Endoscopic ultrasound (EUS) has been used in clinical practice for the past two decades, mainly for staging upper gastrointestinal tract tumours. More recently the technique has been used to guide interventional procedures, particularly EUS guided biopsy. In this review we describe the equipment and technique required for interventional EUS. We also discuss with illustrations the current clinical applications, potential complications and developments for the future. PMID- 12569949 TI - [Saint Panteleimon van Nicomedia, healer]. PMID- 12569950 TI - [Consumption coagulopathy and sepsis]. PMID- 12569951 TI - Current awareness in geriatric psychiatry. Bibliography. PMID- 12569952 TI - [Thrombolytic therapy in acute myocardial infarct]. PMID- 12569953 TI - Diffuse white oral plaques. PMID- 12569955 TI - Bibliography. Current world literature. Urinary tract infections and sexually transmitted diseases. PMID- 12569954 TI - Bibliography. Current world literature. Benign prostatic hyperplasia. PMID- 12569956 TI - [Effect of "home of asthma" action on patients with asthma]. PMID- 12569957 TI - Cranial trauma in adults in physical medicine and rehabilitation: from coma to wakefulness (Short and long texts of jury recommendations). PMID- 12569958 TI - Therapeutic use of botulinum toxins: background and history. AB - The seven botulinum neurotoxin serotypes share less than 50% sequence homology and are immunologically distinct. The neurotoxins inhibit release of the neurotransmitter acetylcholine from the axon terminals of motor neurons, preganglionic sympathetic and parasympathetic neurons, and postganglionic parasympathetic nerves by a multi-step mechanism that differs slightly, but significantly, for each serotype. The inhibition is long lasting but temporary. The resulting muscle paralysis has provided the basis for therapeutic use of botulinum toxin types A and B in a variety of focal dystonias. The safety of the botulinum toxins, when administered focally, has permitted their widespread use in a number of other painful conditions. PMID- 12569959 TI - Possible mechanisms for the effects of botulinum toxin on pain. AB - The therapeutic effects of botulinum toxin are principally, if not exclusively, derived from an alteration in the release of acetylcholine (ACh) at pre-synaptic neurons. The rationale for how these effects could be beneficial in conditions characterized by excessive muscle contraction is clear, but the hypotheses regarding botulinum toxin-induced effects on pain are highly speculative. We explore five possible mechanisms by which botulinum toxin could directly or indirectly alter pain, including: 1) changes in the sensitivity and response patterns of group III and IV muscle nociceptors, 2) diminished activity in the gamma-motor neurons and consequent changes in muscle spindle afferents, 3) alterations in cholinergic control of vascular and autonomic functions, including neurogenic inflammation, 4) induced neuroplastic changes in the processing of afferent somatosensory activity at multiple levels of the neuroaxis, and 5) direct non-cholinergic effects on pain afferents. PMID- 12569960 TI - Considering the immune response to botulinum toxin. AB - The immune system is unable to determine whether material it encounters is deleterious, benign, or even beneficial to the organism. This presents a significant challenge when protein-based biological therapies, such as botulinum toxin, are administered to patients. Many factors combine to influence the likelihood and the magnitude of an immune response if a response is elicited. Those factors intrinsic to antigens that heighten their immunogenicity include nonhuman origin, larger molecules, and aggregated forms of the protein. Extrinsic factors also must be considered, such as the presence of adjuvants in the formulation, either intended or unintended; increasing amounts of antigen within specific dosing ranges; frequent dosing; and, finally, the genetic predisposition of the patient. Once present, not all immune responses preclude the biological therapy from being clinically effective. Only antibodies that bind botulinum toxin in a manner that neutralizes its biological activity will attenuate its effect on the neuromuscular junction. The majority of anti-toxin antibodies do not affect its function. Finally, although crossreactivity has been reported among the seven botulinum toxin serotypes, non-neutralizing antibodies are present that recognize regions of similarity among the serotypes. No cross neutralizing antibodies have been described in patients administered any of the toxin serotypes. PMID- 12569961 TI - Review of the FDA-approved uses of botulinum toxins, including data suggesting efficacy in pain reduction. AB - Botulinum toxin has dramatically improved the treatment of a variety of neurologic disorders. Two botulinum toxin preparations are commercially available in the United States: type A (Botox) and type B (Myobloc). Current indications approved by the United States Food and Drug Administration include cervical dystonia, strabismus, blepharospasm, hemifacial spasm, and glabellar wrinkles for Botox, and cervical dystonia for Myobloc. Botulinum toxin inhibits release of acetylcholine from the neuromuscular junction, resulting in a localized paralysis when minute doses are injected. This mechanism enables botulinum toxin to alleviate symptoms of focal dystonias (which are characterized by excessive muscle contraction), and it may also, along with other theoretical mechanisms, be responsible for pain relief. Studies conducted in patients with cervical dystonia have shown that botulinum toxin effectively reduces pain associated with this disorder, suggesting that this agent may be effective in alleviating other painful syndromes. PMID- 12569962 TI - Botulinum toxin in pain management of soft tissue syndromes. AB - Botulinum toxin is approved for the treatment of muscle overactivity associated with several disorders, such as dystonias. However, control of muscle spasm often results in pain relief as well. Effective relief of pain associated with myofascial pain syndrome provides a model for the use of botulinum toxin to relieve pain associated with other types of soft-tissue syndromes, such as fibromyalgia. Although the mechanisms that trigger the pain in these syndromes vary, recent data suggest that a central neuroplastic mechanism may contribute to many complex pain syndromes. Botulinum toxin therapy may be particularly useful in soft-tissue syndromes that are refractory to traditional treatment with physical therapy, electrical muscle stimulation, and other approaches. Although not used as first-line therapy for pain relief, botulinum toxin may decrease pain long enough for patients to resume more conservative therapy. A primary benefit of treatment with botulinum toxin is its long duration of action. Several studies have demonstrated the efficacy of botulinum toxin types A and B in treating several neuropathic pain disorders. Proper patient selection, injection technique, and dosing are critical to obtaining the best outcomes in managing pain with botulinum toxin. Additional study is needed to better characterize its use for the treatment of pain. PMID- 12569963 TI - A focused review of the use of botulinum toxins for low back pain. AB - Chronic low back pain is the second most common illness reported by patients in the United States and accounts for substantial morbidity and health-care resource utilization. Many back and spine stressors can contribute to tissue injury, resulting in acute or chronic pain. In response to injury, biochemical processes that cause inflammation and nerve sensitization increase pain levels and contribute to a cycle of reactivity that further heightens patients' sensitivity to pain stimuli. Treatment of back pain depends on its severity, duration, and underlying cause. Traditional therapeutic options include exercise, oral anti inflammatory or analgesic medication, antidepressants, physical therapy and, in severe cases, surgery. Unfortunately, dissatisfaction with treatment of back pain is common. Oral medications may not completely alleviate symptoms, and opioid analgesics must be used with caution because of their addictive properties. Surgery does not always produce relief and, in some cases, may even exacerbate the problem. Botulinum toxin, which has already been shown to alleviate pain associated with cervical dystonia and other conditions characterized by muscle spasticity, is now being studied for the treatment of back pain. Preliminary evaluations have shown that this treatment is safe and has the advantage of providing local relief directly to the site of injury or pain, without causing systemic side effects. Initial data from small trials also suggest that botulinum toxin is effective, alleviating back pain in selected patients. On the basis of these promising results, additional study in larger trials is warranted. PMID- 12569964 TI - Use of botulinum toxin in chronic whiplash-associated disorder. AB - Whiplash-associated disorders (WADs) occur as a result of trauma and are often due to motor vehicle accidents and sports injuries. Cervical injury is attributed to rapid extension followed by neck flexion. The exact pathophysiology of WAD is uncertain but probably involves some degree of aberrant muscle spasms and may produce a wide range of symptoms. Initial treatment of pain associated with whiplash usually includes oral medications, such as muscle relaxants and nonsteroidal anti-inflammatory drugs. However, these agents are limited by potential systemic adverse effects. Some patients with chronic WAD may benefit from radiofrequency neurotomy. A new approach to treatment is the use of botulinum toxin, which acts to reduce muscle spasms. Type A toxin (Botox) has been studied in small trials of patients with WAD and has generally been found to relieve pain and improve range of motion. In addition, recent preliminary data from a small trial showed that type B toxin (Myobloc) produced almost immediate pain relief for most patients with post-whiplash headache. Although botulinum toxin has not been evaluated in large long-term trials, these initial data are promising and suggest a role for this agent in the treatment of WAD. Additional study is needed to identify the subset of patients with WAD who are most likely to respond to treatment with botulinum toxin. PMID- 12569965 TI - Use of botulinum toxins for chronic headaches: a focused review. AB - Botulinum toxin has been shown to effectively treat several types of pain associated with neurologic disorders. It has recently been evaluated for the treatment of various types of headaches. In studies of migraine headache, chronic daily headache (more than 15 days of headache per month), tension-type headache, and post-whiplash headache, patients have reported decreased pain after treatment with botulinum toxin type A. A more recently available preparation, botulinum toxin type B, has also been shown to provide relief to patients with transformed migraine headache and post-whiplash headache. Additional study is under way to determine the effectiveness of botulinum toxin for the treatment of chronic cluster headaches. Although the pathophysiology of headache is incompletely understood, muscle tension may trigger or aggravate migraine headaches. Botulinum toxin, which reduces muscle hyperactivity, may reduce headache pain by decreasing muscle tension. It may also provide peripheral and central neurogenic effects and reduce inflammation. Large, rigorously controlled trials of botulinum toxin are needed to better characterize its effects on various types of headaches and its role as a therapeutic agent. Current data suggest that botulinum toxin is safe and does not produce systemic effects associated with other types of headache treatments. PMID- 12569966 TI - A focused review on the use of botulinum toxins for neuropathic pain. AB - Understanding the pathophysiology of a pain syndrome is helpful in selecting appropriate treatment strategies. Nociceptive pain is related to damage to tissues due to thermal, chemical, mechanical, or other types of irritants. Neuropathic pain results from injury to the peripheral or central nervous system. Common examples of neuropathic pain include postherpetic neuralgia, diabetic neuropathy, complex regional pain syndrome, and pain associated with spinal cord injuries. Nociceptive pain may have similar clinical characteristics to neuropathic pain. It is also possible for acute nociceptive pain to become neuropathic in nature, as with myofascial pain syndrome. A clear benefit of botulinum toxin therapy for treatment of neuropathic pain disorders is that it often relieves pain symptoms. Although the precise mechanism of pain relief is not completely understood, the injection of botulinum toxin may reduce various substances that sensitize nociceptors. As a result, botulinum toxin types A and B are now being actively studied in nociceptive and neuropathic pain disorders to better define their roles as analgesics. PMID- 12569967 TI - Treatment of spasticity with botulinum toxin. AB - Spasticity is an abnormal increase in muscle contraction often caused by damage to central motor pathways that control voluntary movement. During clinical examination, spasticity manifests as an increase in stretch reflexes, producing tendon jerks and resistance appearing as muscle tone. There are many causes of spasticity, including demyelination from multiple sclerosis, congenital damage from diseases such as cerebral palsy, trauma to the brain or spinal cord, hemorrhage or infarction, and other pathologic conditions that interrupt neural pathways. Effects of spasticity range from mild muscle stiffness to severe, painful muscle contractures and repetitive spasms that reduce mobility and substantially impede normal activities of daily living. Botulinum toxin therapy reduces spasticity and pain associated with several disorders. Local treatment with botulinum toxins can be used as adjunctive therapy, along with oral antispasticity medications, or alone to provide localized decrease in symptoms of spasticity and pain. Botulinum toxin therapy may be particularly useful for patients with spasticity due to stroke, whose treatment can be tailored based on recovery of function over time. In addition, botulinum toxin therapy is safe for pediatric patients, including children with cerebral palsy, who may not be able to tolerate the cognitive side effects of oral medications. Results of studies evaluating botulinum toxin for the treatment of spasticity due to various causes are presented here. PMID- 12569968 TI - Review of the use of botulinum toxin for hyperhidrosis and cosmetic purposes. AB - This article reviews the current status of dermatological uses for botulinum toxin type A (Botox), recently approved in the United States for treatment of glabellar wrinkles, and type B (Myobloc), approved for cervical dystonia. The respective formulations of Botox and Myobloc are described, and injection techniques and special considerations for administration in the treatment of dermatologic conditions are also discussed. The use of botulinum toxin injections for cosmetic treatment of movement-related facial lines and platysmal bands in the neck is reviewed, including injection procedures, efficacy, and potential complications. Recent developments in the use of botulinum toxins for the treatment of palmar and axillary hyperhidrosis are also described, comparing type A and type B results. Although direct comparisons between botulinum toxins for dermatologic applications are complicated by the lack of functional equivalence of the standard potency assays, appropriate dosing strategies for obtaining satisfactory clinical results using type B are being established, which will add to the experience already gained with type A. The diffusion characteristics of type B appear to show different and potentially advantageous clinical profiles in the treatment of crows' feet and hyperhidrosis compared with type A. PMID- 12569969 TI - Treatment of temporomandibular disorders with botulinum toxin. AB - Temporomandibular disorder (TMD) is a collective term used to characterize a heterogeneous group of conditions involving the temporomandibular joint (TMJ) and its contiguous tissues. Although the pathologies behind TMDs have not been completely explained, the symptoms associated with these disorders are similar and are most commonly manifest as pain in the orofacial region. In preliminary studies, botulinum toxin has been used successfully to treat various pain syndromes, including TMDs. Because of the complex nature of TMDs and proximity of affected muscles to facial nerves, correct injection technique and appropriate dosing guidelines are very important for successful results. This article describes common TMDs and their treatment with botulinum toxin. Dosing guidelines and illustrations of affected muscles and target injection sites are provided. PMID- 12569970 TI - The placenta: a portal of fetal allergen exposure. PMID- 12569971 TI - Products of 15-lipoxygenase: are they important in asthma? PMID- 12569972 TI - Optimizing the use of chromones in the management of asthma: attention to detail over delivered dose and method of administration is essential. PMID- 12569973 TI - Materno-fetal passage of nutritive and inhalant allergens across placentas of term and pre-term deliveries perfused in vitro. AB - BACKGROUND: The pre- and postnatal environment appears to be of crucial importance for the manifestation of allergic diseases, which often begin during infancy. Although T cell reactivity of fetal origin to a range of common allergens is present in most cord blood samples, the immunological basis remains unclear. OBJECTIVE: In order to test the hypothesis of transplacental allergen transfer we studied double-sided open ex vivo perfusion experiments of isolated placental cotyledons with the nutritive allergens beta-lactoglobulin (BLG) and ovalbumin (OVA) and the inhalant major birch pollen allergen Bet v1. METHODS: Placentas of full-term and pre-term newborns were obtained immediately after delivery to recover functionally active maternal and fetal circulations. Thus, a fetal artery and a fetal vein were cannulated and perfused with pure medium (fetoplacental circulation), whereas the intervillous space of placentas was flushed with allergen containing medium by puncture of the basal plate (maternoplacental circulation). Samples that were collected throughout the perfusion experiment from fetal venous outflow were tested by allergen-specific enzyme-linked immunosorbent assays (ELISA) for the presence of allergens indicative of materno-fetal transplacental passage. RESULTS: We observed transplacental transfer of BLG, OVA and Bet v1 in placentas of term as well as premature deliveries. The respective allergen was readily detectable in fetal effluent at the beginning of the perfusion experiment and allergen levels reached a plateau after about 2 h. The steady state transfer rate of BLG and OVA in term placentas was 0.012% +/- 0.001 and 0.013% +/- 0.001 of total dose, i.e. 130.21 +/ 7.41 ng/mL and 115.83 +/- 6.07 ng/mL, respectively. The observed transfer rate of Bet v1 after 2h of perfusion was 0.155% +/- 0.034 of total dose, that is 2.41 +/- 1.36 ng/mL. Transplacentally transferred concentration of BLG and OVA in pre term placentas increased continuously throughout perfusion time from 5.32 +/- 0.92 ng/mL at 1 min to 87.53 +/- 21.93 ng/mL at 120 min and 1.35 +/- 0.31 ng/mL at 1 min to 112.87 +/- 5.25 ng/mL at 150 min, respectively. CONCLUSION: Allergen specific cord blood reactivity may be attributed to low levels of allergens crossing the human placenta and providing the fetus with the necessary stimulus for T cell priming. PMID- 12569974 TI - Polyclonal and allergen-induced cytokine responses in children with elevated immunoglobulin E but no atopic disease. AB - BACKGROUND: Reduced Th1 and elevated Th2 cytokine responses are considered to be a principal mechanism in the generation of the inflammation leading to the manifestations of atopic disease in the skin of atopic dermatitis and in the airways of asthma. If reduced Th1 and elevated Th2 responses are principal determinants of the manifestation of atopic disease it might be expected that subjects with established disease would exhibit differences in their cytokine profiles as compared with atopic patients without clinical disease. OBJECTIVE: To determine whether asymptomatic atopic children exhibit a cytokine imbalance similar to that seen in patients with established atopic disease or if they behave like non-atopic controls. Cytokine responses in a group of children with elevated IgE but no clinical manifestations of disease, atopic children with established disease and non-atopic controls were compared. METHODS: We examined allergen-induced (house dust mite, HDM, rye grass pollen and RYE) cytokine responses in parallel with polyclonal (staphylococcal enterotoxin B, SEB) cytokine responses in a group of children with elevated serum IgE levels without current or past evidence of atopic disease (median age 6.6 years) and compared these with a non-atopic control group (median age 6.5 years) and a group of children with atopic disease (median age 6.7 years). RESULTS: Symptomatic atopic children had reduced SEB-induced IFN-gamma and increased SEB-induced IL-4 and IL 5 as compared with non-atopic controls. In contrast, SEB-induced IFN-gamma, IL-4 and IL-5 production in asymptomatic atopics was not significantly different from the non-atopic control subjects. Allergen-induced Th1 (IFN-gamma) and Th2 (IL-5 and IL-13) cytokine production was increased in both symptomatic atopics and asymptomatic atopics when compared with non-atopic controls. CONCLUSION: The defect in polyclonally induced IFN-gamma production was associated with the clinical manifestation of atopic disease but not the atopic stateper se. This suggests that the global reduction in IFN-gamma is the key determinant of the development of overt atopic disease. In contrast, elevated allergen-induced Th2 cytokine responses in children related to the atopic state per se irrespective of the presence of clinical atopic disease. PMID- 12569975 TI - Expression and activation of 15-lipoxygenase pathway in severe asthma: relationship to eosinophilic phenotype and collagen deposition. AB - BACKGROUND: Although 15(S)-hydroxyeicosatetraenoic acid (15(S)-HETE), a product of 15-lipoxygenase (15-LO), may be involved in mild to moderate asthma, little is known about its potential roles in severe asthma. OBJECTIVES: This study was performed to evaluate 15(S)-HETE levels in bronchoalveolar lavage fluid (BALF) from severe asthmatics with and without airway eosinophils and from the control groups. In addition, 15-LO protein expression was examined in endobronchial biopsy, while its expression and activation were evaluated in BAL cells. RESULTS: While 15(S)-HETE levels in BALF were significantly higher in all severe asthmatics than normal subjects, severe asthmatics with airway eosinophils had the highest levels compared with mild, moderate asthmatics and normal subjects. 15(S)-HETE levels were associated with tissue eosinophil numbers, sub-basement membrane thickness and BALF tissue inhibitor of metalloproteinase-1 levels, and were accompanied by increased 15-LO expression in bronchial epithelium. In addition, activation of 15-LO was suggested by the increased proportion of 15-LO in the cytoplasmic membrane of alveolar macrophages from severe asthmatics. CONCLUSION: The data suggest that severe asthmatics with persistent airway eosinophils manifest high levels of 15(S)-HETE in BALF, which may be associated with airway fibrosis. It is likely that 15-LO expression and activation by airway cells explain the increased 15(S)-HETE levels. PMID- 12569976 TI - Uncoupled regulation of leukotriene C4 synthase in platelets from aspirin intolerant asthmatics and healthy volunteers after aspirin treatment. AB - BACKGROUND: We have reported that thromboxane A2 induces suppression of leukotriene (LT) C4 synthase activity in human platelets. AIM: In the present study, we describe a mechanism whereby aspirin treatment can lead to increased formation of LTC4, which is a potent bronchoconstrictor and inflammatory mediator. This mechanism is also demonstrated to be present in platelets from aspirin-intolerant asthmatics (AIA). METHODS: The effect of arachidonic acid or platelet agonists on LTC4 synthase activity was investigated in platelets obtained from healthy volunteers, aspirin-intolerant asthmatics or aspirin tolerant asthmatics after in vivo treatment or in vitro pre-incubation with aspirin. RESULTS: Incubation of normal platelets with arachidonic acid or collagen provoked approximately 50% reduction of platelet LTC4 synthase activity, as determined by the conversion of LTA4 to LTC4. However, the inhibitory effect of arachidonic acid or collagen was not observed after oral administration of aspirin prior to collection of the platelets. Arachidonic acid-induced inhibition of LTC4 synthase activity was totally abolished in platelets collected from peripheral blood already 30 min after aspirin ingestion but was fully restored in platelets collected 3 to 7 days after the administration of aspirin. Treatment of platelet suspensions with aspirin in vitro dose-dependently counteracted the suppressive effect of arachidonic acid on LTC4 formation, with total reversal at approximately 40 microm. In contrast, the major aspirin metabolite, salicylic acid did not alter arachidonic acid-induced reduction of LTC4 synthase activity. Similarly, LTC4 synthase activity in platelets from AIA and aspirin-tolerant asthmatics (ATA) was reduced by approximately 50% after pre-treatment with arachidonic acid in vitro. Again the inhibitory effect was abolished when platelets were pre-incubated in the presence of aspirin. CONCLUSION: The results indicate that oral aspirin administration can lead to uncoupling of thromboxane A2-dependent negative feedback mechanisms, which may normally restrict the production of cysteinyl leukotrienes. This mechanism can be of potential interest in aspirin-induced asthma. PMID- 12569977 TI - Liposome-entrapped D. pteronyssinus vaccination in mild asthma patients: effect of 1-year double-blind, placebo-controlled trial on inflammation, bronchial hyperresponsiveness and immediate and late bronchial responses to the allergen. AB - BACKGROUND: Allergen vaccination is effective in mite-allergic asthma. Liposomes are immunological adjuvants that can act as allergen carriers. OBJECTIVE: To evaluate the immunological and functional effects of a liposome-entrapped D. pteronyssinus vaccine on mite monosensitive, mild asthma patients. METHODS: A double-blind, placebo-controlled trial was conducted on 26 asthma patients who randomly received vaccination or placebo for 1 year. The levels of exposure to Der p 1 allergen were constant during the study. Allergen bronchial challenge was made at the beginning (T0) and after 1 year of treatment (T12). The day before and 24 h after the allergen provocation, patients were challenged with methacholine (Mth) (until FEV1 fell by 40%) and blood and sputum samples were obtained. Dose-response curves to Mth were evaluated in terms of Mth-PD20 (dose of Mth that induced 20% drop in FEV1), slope (Mth-DRS) and level of plateau. Blood and sputum eosinophils and serum levels of eosinophil cationic protein (ECP) and intercellular adhesion molecule-1 (ICAM-1) were measured. RESULTS: Groups were comparable at the start of the trial. At TI2, previous to the allergen challenge, the active group showed higher values of both FEV1 and Mth PD20 and lower values of Mth-DRS. The number of patients presenting a level of plateau increased in the active group (from two to four) and decreased in the placebo group (from two to one). At T12, before the allergen challenge, serum ECP levels increased in the placebo group and blood eosinophils showed a trend towards lower numbers in the active one. The immediate response and the changes in Mth-DRS values, sputum eosinophils and serum ECP levels following the allergen challenge were attenuated in the active group. CONCLUSION: Liposome-entrapped D. Pteronyssinus vaccination: (i) protects mild asthma patients from the worsening of asthma due to sustained mite exposure; and (ii) reduces the functional and inflammatory changes induced by allergen bronchial provocation. PMID- 12569978 TI - Phage-displayed Bet mim 1, a mimotope of the major birch pollen allergen Bet v 1, induces B cell responses to the natural antigen using bystander T cell help. AB - BACKGROUND AND OBJECTIVE: In previous studies we have generated mimotopes of Bet v 1, the major birch pollen allergen, by biopannings of phage-display random peptide libraries. In the present study, we analysed the humoral and cellular immune response to Bet v 1-mimotopes. METHODS: The mimotope CFPYCYPSESA, designated Bet mim 1, was used for intraperitoneal immunizations of BALB/c mice in phage-displayed form. For examination of the humoral immune response, enzyme linked immunosorbent assay (ELISA) experiments were applied. Stimulation capacities were investigated in cultured mouse splenocytes and in humoral Bet v 1 specific T cell clones. RESULTS: We demonstrated that the Bet mim 1-induced murine antibody response against Bet v 1 was predominated by the IgG1 isotype. In these mice only the phage-displayed mimotopes, but neither the allergen nor the synthetic Bet mim 1-mimotopes were able to stimulate proliferation of cultured splenocytes. Using Bet v 1-specific T cell clones of allergic patients, phage displayed and synthetic mimotopes were unable to stimulate T cell proliferation. Moreover, tolerance induction to Bet v 1 in mice by intranasal administration of Bet mim 1-phages or Bet mim 1-peptide failed. CONCLUSION: Taking these results together, our data indicate that Bet mim 1 mimics a Bet v 1-epitope on the B cell but not on the T cell level. We suggest that the phage itself is responsible for the recruitment of T cells providing bystander help in the formation of a mimotope-specific humoral response. PMID- 12569979 TI - T cell cytokine responses to outer membrane proteins of Haemophilus influenzae and the house dust mite allergens Der p 1 in allergic and non-allergic subjects. AB - BACKGROUND: Haemophilus influenzae are ubiquitous colonizers of the nasopharynx, Little is known about the T cell cytokine responses to the antigens of these bacteria and whether or not the responses may interact with responses to aeroallergen. OBJECTIVE: To measure the T cell cytokine responses to conserved outer membrane protein antigens of Haemophilus influenzae and to house dust mite allergen of subjects allergic to the house dust mite and of subjects without allergic sensitization. METHODS: T cell responses were measured by in vitro proliferation and cytokine release from peripheral blood monocytes (PBMC). The allergen used was Der p 1 and outer membrane proteins were recombinant polypeptides representing the OMP6 and D15 antigens. RESULTS: The PBMC of most subjects had proliferative responses to OMP6 and D15, which were highly correlated. The pattern of cytokine release was Th1 biased with high levels of IFN-gamma and usually little IL-5 or IL-13 although PBMC from a few subjects did release IL-5 independent of allergic status. IL-10 release was readily detected. There was no difference in the anti-OMP cytokine response of PBMC from subjects without any known allergy and the responses of PBMC from subjects who were highly allergic to house dust mite. The responses to the Der p 1 allergen showed the expected Th2 cytokine release. CONCLUSION: The outer membrane protein antigens of the ubiquitous colonizing bacteria Haemophilus influenzae induce Th1 cytokine responses which are similar for PBMC from non-allergic individuals and subjects with a high degree of allergy to the perennial house dust mite allergen and strong Th2 responses to Der p 1. PMID- 12569980 TI - A rapid test for detection of mite allergens in homes. AB - BACKGROUND: International guidelines recommend allergen avoidance for asthma management, but do not include making assessments of allergen exposure. Mite allergen exposure cannot be assumed, especially in geographical regions where climatic conditions vary. OBJECTIVE: To develop a rapid test that would enable consumers to detect mite allergen in the home. METHODS: A lateral flow test using gold labelled antibody for mite group 2 allergen was developed as part of a detection kit incorporating the MITEST dust sampling device. Dust samples were assayed by ELISA for group 1 and group 2 allergens and by using the rapid test. The tests were compared as indices of mite allergen exposure. RESULTS: There was a good correlation between group 1 and group 2 levels by ELISA (n = 349, r = 0.60, P < 0.001). In a multi-centre study of 65 homes (263 dust samples) in five countries, there was a strong correlation between ELISA and the rapid test. Most samples with high scores in the test (43/48, 90%) contained > 1 microg/m2 group 2 allergen, whereas most low samples contained < 1 microg/m2 (50/64, 78%). Differences between mean group 2 levels of samples that scored low (0.28 microg/m2), medium (1.68 microg/m2) or high (3.18 microg/m2) on the test were highly significant (P 0.007 to < 0.001). CONCLUSIONS: A simple rapid test has been developed that detects mite allergen in the home within 10 min. The mite screening test should educate consumers about allergen exposure and encourage compliance with allergen-avoidance procedures. This technology has applications for the detection of other common environmental allergens. PMID- 12569981 TI - Allergens, Der p 1, Der f 1, Fel d 1 and Can f 1, in newly bought mattresses for infants. AB - BACKGROUND: To avoid allergen exposure of newborn babies, the use of a new mattress for the baby bed may be recommended. However, it is not certain that new mattresses are always free of allergens. OBJECTIVE: In the present study the allergen content of new infant mattresses was investigated. METHODS: Dust samples were vacuumed from 90 new mattresses for infant beds bought in 50 different Dutch shops, and the concentrations of Der p 1, Der f 1, Fel d 1 and Can f 1 were determined by radioimmunoassays. RESULTS: Most mattresses contained some allergen and often the allergen concentrations were surprisingly high. Only 15 of the 90 mattresses contained no detectable amounts of any of the four allergens. The highest concentration found for each allergen was 3.1, 46.5, 20.2 and 95.7 microg/g of dust, respectively. However, the total amount of allergen in a mattress was still rather low because the new mattresses contained only modest amounts of dust. Baby mattresses more often contained an increased allergen load than the larger, standard-sized, infant mattresses. This may be caused by differences in manufacturing procedure. Also, mattresses that were sold without a plastic encasement more often contained an increased allergen load. CONCLUSIONS: It is advisable to buy a mattress that is wrapped in plastic, but it may still contain a substantial amount of allergen. Thorough vacuuming of a newly bought mattress before it is installed on a child's bed, is also advisable. After instalment, regular vacuuming of the mattress and washing of the bed linen as well as measures to eliminate allergen reservoirs in other parts of the house are important, because our observations indicate that mattresses easily pick up allergens from the environment. PMID- 12569982 TI - The use of a neural network to forecast daily grass pollen concentration in a Mediterranean region: the southern part of the Iberian Peninsula. AB - BACKGROUND: Pollen allergy is a common disease causing hayfever in 15% of the population in Europe. Medical studies report that a prior knowledge of pollen content in the air can be useful in the management of pollen-related diseases. OBJECTIVES: The aim of this work was to forecast daily Poaceae pollen concentrations in the air by using meteorological data and pollen counts from previous days as independent variables. METHODS: Linear regression models and co evolutive neural network models were used for this study. Pollen was monitored by a Hirst-type spore trap using standard techniques. The data were obtained from the Spanish Aerobiology Network database, University of Cordoba Monitoring Unit. The set of data includes a series of 20 years, from 1982 to 2001. A classification of the years according to their allergenic potential was made using a K-mean cluster analysis with pollen and meteorological parameters. Statistical analysis was applied to all the years of each class with the exception of the most recent year, which was used for model validation. RESULTS: It was observed that cumulative variables and pollen values from previous days are the most important factors in the models. In general, neural network equations produce better results than linear regression equations. CONCLUSION: Co evolutive neural network models, which obtain the best forecasts (an almost 90% "good" classification), make it possible to predict daily airborne Poaceae pollen concentrations. This new system based on neural network models is a step toward the automation of the pollen forecast process. PMID- 12569983 TI - Long-range destruction of Der p 1 using experimental and commercially available ionizers. AB - BACKGROUND: To reduce the risk of sensitization and the elicitation of allergy symptoms, it is important to reduce the level of allergens in the home. It has previously been demonstrated that corona discharge, the process by which ionizers produce ions, can destroy the major house dust mite allergen Der p 1. OBJECTIVE: In this paper the denaturing efficacy of an experimental ionizer and two commercially available products are evaluated. METHODS: The first test was conducted in an electrically grounded chamber with samples of Der p 1 placed in various positions for 1, 2 and 3 weeks. The second test was conducted in situ in an unoccupied, furnished office room for 1 week. Der p 1 concentration was quantified by two-site monoclonal antibody enzyme-linked immunosorbent assay (ELISA). RESULTS: All ionizers in both tests caused significant reductions in allergen concentration (P < 0.05), reaching a maximum of 92% with the experimental ionizer in the chamber after 3 weeks. The percentage reductions observed in situ with the experimental and the larger commercial ionizer were similar, reaching a maximum of 32% at a distance of 4 m away from the experimental ionizer after 1 week of exposure. CONCLUSION: With a revised protocol for use, air ionizers may offer a simple, efficient and inexpensive way to reduce allergen levels in the domestic environment. PMID- 12569984 TI - Comparison between the native glycosylated and the recombinant Cup a1 allergen: role of carbohydrates in the histamine release from basophils. AB - BACKGROUND: Cypress pollinosis is an important cause of respiratory allergies. Recently, the Cupressus arizonica major allergen, Cup a1, has been cloned and expressed. The native counterpart of this allergen has been purified and characterized by our group. It has been suggested that sugar moieties play a role in the in vitro IgE binding on Cupressus arizonica pollen extract. OBJECTIVE: To characterize the immunoreactivity of the recombinant major allergen in comparison with its native counterpart. To evaluate the role of carbohydrate moieties in the IgE-mediated in vitro histamine release from basophils by using the native glycosylated Cup a1 as compared with the recombinant one. METHODS: Recombinant Cup a1 was expressed in E. coli. IgE reactivity of Cupressaceae-allergic patients on the native as well as the recombinant molecule was investigated by immunoblotting, ELISA experiments and histamine release test from passively sensitized basophils. RESULTS: Fourteen out of 17 Cup a1-positive sera had IgE antibodies reactive with the native molecule only and lost their reactivity-after periodate deglycosylation of the allergen. Moreover, only native molecule was capable of inducing histamine release by this group of sera. Both the recombinant and the native molecules were recognized by three out of the 17 sera and were equally capable of triggering degranulation. CONCLUSION: A large number of sera reactive with the major allergen recognize carbohydrate epitopes only. IgE from these sera are able to induce histamine release from basophils and they might play a functional role in the clinical symptoms of allergy. PMID- 12569985 TI - Studies on the carbohydrate moiety of Pla l 1 allergen. identification of a major N-glycan and significance for the immunoglobulin E-binding activity. AB - BACKGROUND: Pla l 1, the major allergen of Plantago lanceolata pollen, is a glycoprotein that contains an N-glycosylation site. Carbohydrate moieties of many allergenic glycoproteins have been reported to be IgE-binding determinants responsible for cross-reactivity among different species. OBJECTIVE: To identify the kind of linkages and the type of glycans present in Pla l 1 and to investigate their contribution to the allergic response to this allergen. METHODS: Pla l 1 was deglycosylated by N-glycosidase A and the IgE-binding ability of the unglycosylated protein was evaluated by dot-blot. Identification of beta1 --> 2 xylose and/or alpha1 --> 3 fucose residues in Pla l 1 N-glycan was carried out by incubation with specific antibodies from rabbit antiserum against HRP (anti-HRP). The contribution of this N-glycan to total IgE reactivity was analysed quantitatively by pre-incubation of Pla l 1 with anti-HRP prior to incubation with sera. The role of the carbohydrate moiety of Pla l 1 in cross reactivity was studied by RAST using unrelated glycoproteins with known sugar composition and structure. RESULTS: The effectiveness of N-glycosidase A to deglycosylate Pla l 1 and the ineffectiveness of the treatment with PNGase F indicate that Pla l 1 carries a complex N-glycan with an alpha1 --> 3 fucose residue in its structure. Furthermore, the presence of beta1 --> 2 xylose and/or alpha1 --> 3 fucose residues was identified in this N-glycan by means of an ELISA. Pre-incubation of Pla l 1 with an anti-HRP antibody caused a weak but significant reduction in IgE reactivity. Some sera from P. lanceolata-allergic patients reacted positively with four glycoproteins that bear N-glycans of complex type but not with fetuine. CONCLUSIONS: Pla l 1 is a glycoprotein that carries at least a complex, major N-linked glycan, with a alpha1 --> 3 fucose residue in its structure and probably also a beta1 --> 2 xylose. This glycan moiety does not seem to constitute a relevant allergenic epitope of Pla l 1. PMID- 12569986 TI - Non-covalent presentation of sulfamethoxazole to human CD4+ T cells is independent of distinct human leucocyte antigen-bound peptides. AB - BACKGROUND: It has been shown that drugs comprise a group of non-peptide antigens that can be recognized by human T cells in the context of HLA class II and that this recognition is involved in allergic reactions. Recent studies have demonstrated a MHC-restricted but processing- and metabolism-independent pathway for the presentation of allergenic drugs such as lidocaine and sulfamethoxazole (SMX) to drug-specific T cells. However, there is little information so far on the precise molecular mechanisms of this non-covalent drug presentation. OBJECTIVE: The aim of this study was to evaluate the requirements for a specific peptide occupying the groove of the MHC class II molecule for the efficient presentation of non-covalently bound drugs to CD4+ T cells. METHODS: We analysed the effect of coincubation or prepulse of antigen presenting cells (APC) with different peptides on the proliferative responses of SMX-specific CD4+ T cell clones. In a second series of experiments, we eluted HLA-bound peptides from the surface of antigen presenting cells by mild acid treatment. Successful removal of peptides was tested directly using labelled peptides and functionally by monitoring activation and proliferation of peptide-specific T cell clones. Finally, the presentation of SMX to SMX-specific T cell clones before and after elution of MHC class II bound peptides was tested. RESULTS: We found that neither peptide coincubation nor peptide prepulse of APC altered the proliferative response of SMX-specific T cells. APC treated with the acid for a short time retained cell viability, MHC class II expression and antigen presenting cell function. However, defined peptides could be eluted from surface MHC class II molecules nearly quantitatively. Nevertheless, the chemically non-reactive drug SMX could still be presented to specific T cells independent of the presence of distinct self-peptides. CONCLUSION: Our data suggest that small molecules like drugs can bind to a multitude of HLA-bound peptides or that, similar to superantigens, they might bind directly to HLA. PMID- 12569987 TI - beta-Lactam allergenic determinants: fine structural recognition of a cross reacting determinant on benzylpenicillin and cephalothin. AB - BACKGROUND: An appreciation of the structural heterogeneity of allergenic determinants on penicillins and cephalosporins reveals the importance of side chain groups and their involvement in many allergies to beta-lactam drugs. Although allergenic cross-reactions between penicillins and cephalosporins are known to occur, the precise molecular bases of such recognitions and cross sensitivities have rarely been studied and identified. OBJECTIVES: The unexpected finding of a high incidence of positive IgE antibody reactions with both benzylpenicillin and cephalothin prompted serological and immunochemical studies to identify the chemical basis of antibody recognition of these drugs from the two different families of beta-lactam antibiotics. METHODS: Adsorption studies were employed to identify whether or not a single population of antibodies was involved in the recognition of benzylpenicillin and cephalothin. Identification of the fine structural features recognized by IgE antibodies was investigated by quantitative hapten inhibition studies employing carefully selected beta-lactam drugs, analogues and some other structurally related chemicals. RESULTS: Adsorption studies with penicilloic acid-solid phase clearly established that a single population of cross-reacting antibodies recognized both benzylpenicillin and cephalothin. Quantitative inhibition findings, especially with phenylacetic acid and 2-thiopheneacetic acid and with cephaloridine and cefoxitin, which have the same (2-thienyl)methyl side-chain as cephalothin, implicated the methylene group as the focus of the allergenic determinant recognized on benzylpenicillin and cephalothin. In addition to the methylene group, recognition graded into neighbouring structures including the amide group and extended weakly to the beta lactam ring. CONCLUSIONS: Results confirmed that structural features as small as a methylene group may be allergenically important. In the present case, this group, making up only part of the different side-chains on benzylpenicillin and cephalothin, together with neighbouring structures extending toward the beta lactam ring, accounted for the cross-reactivity seen between structures that, at first sight, appear to be not closely related. Such subtle, small, common structural features are likely to be immunologically recognized and implicated in allergic reactions to other drugs, including beta-lactam antibiotics. PMID- 12569988 TI - Exogenous interleukin-16 inhibits antigen-induced airway hyper-reactivity, eosinophilia and Th2-type cytokine production in mice. AB - BACKGROUND: IL-16 has been described as a natural soluble CD4-ligand with immunosuppressive effects in vitro. However, little is known about the effect of IL-16 on immune responses in vivo. OBJECTIVE: In the present study, we examined the effect of IL-16 administration in a murine model of allergic asthma. Next, we determined whether these effects were mediated by modulation of CD4+ T lymphocytes. METHODS AND RESULTS: Intraperitoneal administration of IL-16 completely inhibits antigen-induced airway hyper-responsiveness and largely decreases the number of eosinophils in bronchoalveolar lavage fluid (> 90%) and airway tissue of ovalbumin-sensitized and challenged mice. Firstly, it appears that thoracic lymph node cells isolated from in vivo IL-16-treated ovalbumin challenged animals produce less IL-4 (77%) and IL-5 (85%) upon antigenic re stimulation, when compared to vehicle-treated mice. Secondly, pre-incubation of lymphocytes with IL-16 in vitro reduces antigen-induced proliferation (55%) and Th2-type cytokine production (IL-4; 56%, IL-5; 77%). Thirdly, the presence of IL 16 during priming cultures of TCR transgenic T cells (DO11.10), reduces IL-4 (33%) and IL-5 (35%), but not IL-10 and IFNgamma levels upon re-stimulation. CONCLUSION: It can be concluded that IL-16 has potent immunosuppressive effects on a Th2dominated allergic airway response. PMID- 12569990 TI - Oral tolerance induction to Dermatophagoides pteronyssinus and Blomia tropicalis in sensitized mice: occurrence of natural autoantibodies to immunoglobulin E. AB - BACKGROUND: The dust mites Dermatophagoides pteronyssinus (Dp) and Blomia tropicalis (Bt) are important sources of indoor allergens in tropical and subtropical countries. Murine models allow the analysis of the immune response and regulation of IgE production to Dp and Bt allergens. Oral tolerance induces unresponsiveness in naive animals, but its application in sensitized animals can provide useful information to improve allergy therapy. OBJECTIVE: To study the profile of IgE and IgG subclasses antibody upon oral administration with Bt and Dp extract in previously sensitized mice. Further, the occurrence of autoantibodies IgG anti-IgE in the immunization and in the oral tolerance was investigated. METHODS: A/Sn mice were immunized with Bt or Dp extract in alum, orally administrated with 0.25 mg of Bt or Dp extract or PBS at the 6th, 7th and 8th days after immunization and boosted twice with their respective allergens. To analyse the mice groups, specific IgE antibodies were measured by passive anaphylaxis reaction and specific IgG subclasses and anti-IgE IgG autoantibody by ELISA assay. RESULTS: IgE levels were markedly increased in Bt-immunized mice compared with Dp-immunized mice. A distinct profile of the specific isotypes was verified in Bt-immunized mice with a preferential production of IgG3 and IgA antibodies, whereas Dp-immunized mice developed high titres of anti-Dp IgG1, IgG2a and IgG2b antibodies. The antigen feeding inhibited IgE response in both fed-mice groups but only Dp-fed mice presented decreased levels of IgG antibodies. Free anti-IgE IgG autoantibodies were detected mainly in the Dp immunization and they correlated with the antibody isotypes found against the allergen. CONCLUSIONS: This is the first time that the murine-type I hypersensitivity is employed to study Bt-immunization, showing a marked IgE production, associated with IgG response, which is at least in part driven by T independent antigens. The oral tolerance protocol in previously sensitized animals was able to down-modulate IgE response and points out this route as a strategy for allergy therapy. PMID- 12569989 TI - Eosinophilic inflammation and airway hyper-responsiveness are profoundly inhibited by a helminth (Ascaris suum) extract in a murine model of asthma. AB - BACKGROUND: The increase of atopic disorders in developed countries has been associated with the decline of infectious diseases, including helminthic infections. We have already demonstrated that adult worm extracts from Ascaris suum (ASC) suppress the IgE antibody production against unrelated antigens. OBJECTIVE: Here we investigated the influence of ASC on the development of pulmonary eosinophilic inflammation in a murine model of asthma. METHODS: Heat coagulated egg white alone (EWI) or mixed with ASC (EWI + ASC) was implanted subcutaneously in B10.A or C57BL/6 mice, and 14 days later they were challenged intratracheally with OVA or exposed to aerosolized OVA for 4 days. RESULTS: The suppressive effect of ASC was demonstrated on the accumulation of cells into airways, with reduction of eosinophil numbers and of eosinophil peroxidase activity in EWI + ASC-immunized mice. This effect correlated with a marked reduction of IL-5 and IL-4 levels in the BAL from C57BL/6 and B10. A mice, respectively, and of eotaxin in BAL and lung tissue from both strains. OVA specific IgG1 and IgE levels were also impaired in serum and BAL from these mice. Airway hyper-reactivity to methacholine was obtained in B10. A mice sensitized with EWI, but the respiratory mechanical parameters returned to normal levels in EWI + ASC-immunized mice. CONCLUSION: These results indicate that ASC has a profound inhibitory effect on lung inflammation and hyper-responsiveness and that suppression of IL-5 or IL-4 and of eotaxin contributes to this effect. PMID- 12569991 TI - Contribution to hospital performance: market orientation vs. marketing effort and lack of competition. AB - Marketing is still viewed with some skepticism by some hospital administrators who wonder if marketing is needed when the hospital is in a benign competitive environment. This research seeks to investigate the contribution of a marketing orientation to hospital performance beyond what can be achieved by merely spending money on promotion or not facing stiff competition. Findings reveal that having an authentic market orientation makes a significant contribution to a hospital's success above what can be achieved through promotional budgets and lack of competition. PMID- 12569992 TI - Nursing presence, patient satisfaction, and the bottom line. AB - Hospital planners rely on a number of motivational models to increase patient satisfaction. Missing is the user friendly model oriented to nursing presence. Absent is the approach which taps into that contribution particular nurses might make to increase patient satisfaction and enhance the bottom line. This paper remedies this circumstance by the introduction of the Nursing Presence Grid model. The model is based upon a review of the literature conducted in the manner of Polit and Hungler (1995) and appearing since 1983. Nursing presence is defined. The Nursing Presence Grid matches the various dimensions of nursing presence with various courses of action thought to attain nursing presence. An explanation of how the model might be applied is presented. PMID- 12569993 TI - Nursing: the lost commodity in managed care or marketing health care agencies and the professional nursing shortage. AB - It is essential for Hospital organizations today to have an in depth appreciation for Marketing strategies and practices when dealing with their patients. The most effective first line of defense and action to ensure the highest level of health care service and value for consumers are nurses. This article will explore the essential sophisticated business and marketing role that nurses can play to achieve patient satisfaction. PMID- 12569994 TI - Marketing strategies of hospital service organizations in Nigeria: a study of selected privately owned hospitals in Port Harcourt. AB - This study was centered on the identification of the marketing strategies of hospitals and the factors that militate against their effective adoption and implementation. A total of 80 hospitals in Port Harcourt responded to a structured questionnaire administered on them. It was found, among other things, that many hospital organizations apply marketing strategies, and that hospitals that adopt effective marketing strategies perform better than those that do not. Some of the factors militating against the effective adoption and implementation of marketing strategies include lack of planning, lack of top management support, and non-utilization of all the promotional mix elements. PMID- 12569995 TI - Hospitals and reengineering. AB - One of the key success factors for reengineering in hospitals will be the ability of health care leaders to redefine their organizations in terms of process. Hospital managers are responding with anything from mergers to minor shifts in strategic thinking. Some hospital leaders answering the call for significant change are faced without dated models that are complex and inflexible. PMID- 12569996 TI - Technology and the sales force: Amach, Inc. AB - This paper explores the application of sales force technology by Amach, Inc. a Detroit based medical device firm. Sales force technology represents a variety of dynamic tools that can help the sales force. While these tools cannot replace the salesperson and generate deals from customers, they can enhance efficiency and effectiveness levels. Sales force technology can range from computer laptops, pagers, cellular phones, desktop personal computers and software for contact management, database management and multimedia presentations. The key for sales managers to remember is that these tools do not drive themselves and still need the skill of management to get the best results. Managerial suggestions and research ideas are presented. PMID- 12569997 TI - What can Johnson & Johnson do to remain a giant in the health care industry? AB - As a major Fortune 500 corporation and manufacturer of significant drug products for the pharmaceutical industry, Johnson & Johnson has also had its share of marketing crisis, including the classic case example of The Tylenol Scare in Fall, 1982, so they can appreciate the need for effective marketing performance and customer responsiveness. This article will examine how Johnson & Johnson has adapted to a highly volatile business environment and how they can be benchmarked for highly competitive marketing strategies and practices. PMID- 12569998 TI - Ophthalmic surgery, lasers & imaging. PMID- 12569999 TI - A comparison of quantitative mapping and stereoscopic fundus photography grading of retinal thickness in diabetic eyes with macular edema. AB - OBJECTIVE: To evaluate the ability of a retina specialist's grading of 30 degrees color stereoscopic fundus photographs to identify areas of significant retinal thickening as assessed by the Retinal Thickness Analyzer (RTA) and to determine whether this ability was affected by the presence of retinal pathology. MATERIALS AND METHODS: Thirty-two eyes in 29 patients clinically diagnosed as having diabetic macular edema underwent RTA scanning and nonsimultaneous 30 degrees color stereoscopic fundus photography. Retinal thickness maps of the macular area were generated, and regions with significant retinal thickening (> or = 2 SD above normal values) were identified. A retina specialist reader, masked to the RTA measurements, identified areas with macular edema on the stereoscopic fundus photographs, which subsequently were overlaid on the retinal thickness maps. The sensitivity (percent of significant retinal thickening areas identified by the retina specialist grading the stereoscopic fundus photographs) was calculated separately for areas with and without retinal pathology. Specificity of the stereoscopic fundus photograph grading was assessed similarly. RESULTS: The retina specialist's stereoscopic fundus photography grading identified 78.8% of areas with significant retinal thickening (range over eyes: 20.4%-100%) and was slightly more likely to identify significant retinal thickening when pathology was present (89.6%) than when pathology was not present (78.4%; pooled risk ratio, 1.14 [95% CI = 0.54, 2.42]). Specificity of stereoscopic fundus photography grading was 58%, ie, 42% of areas without significant retinal thickening were (incorrectly) identified as edematous by the stereoscopic fundus photograph grading. This misidentification was more likely if pathology was present (76.9%) than if pathology was not present (41.1%; pooled risk ratio, 1.87 [95% CI = 1.28, 2.73]). CONCLUSION: This study shows the determination of macular edema by a retina specialist reading color stereoscopic fundus photographs is sensitive but not specific with reference to edema identified by the RTA. Furthermore, the presence of retinopathy tends to cause false-positive readings with reference to edema identified by the RTA. These findings indicate the need to use objective, quantitative methods in clinical studies to detect and monitor macular edema. PMID- 12570000 TI - Oral fluorescein angiography in patients with choroidal neovascularization and macular degeneration. AB - OBJECTIVE: To evaluate image quality of oral fluorescein angiography in patients with age-related macular degeneration and choroidal neovascularization. MATERIALS AND METHODS: Twenty four patients with proven choroidal neovascularization were examined in a prospective observational case series. Confocal oral and intravenous fluorescein angiography were performed using the Heidelberg Retina Angiograph. Oral and intravenous image quality were compared in a masked fashion. RESULTS: Choroidal neovascularization could be diagnosed in all patients using intravenous angiograms. Oral fluorescein angiography accurately detected the presence or absence of choroidal neovascularization in all patients and could determine the classification of choroidal neovascularization, location, and extent and borders in 20 (83.3%) of 24 patients. Early, mid, and late phases of oral angiograms occurred at 8, 15, and 26 minutes, respectively. CONCLUSION: Oral fluorescein angiography using the confocal Heidelberg Retina Angiograph allowed detection of choroidal neovascularization in all patients in this study. Visualization of extent and type of choroidal neovascularization was possible in most eyes with choroidal neovascularization. These findings suggest oral angiography is an excellent screening tool for choroidal neovascularization and allows guidance of treatment in the majority of cases. PMID- 12570001 TI - Correlation of clinical and optical coherence tomography findings in postoperative macular hole closure status. AB - OBJECTIVE: To determine the correlation between clinical and optical coherence tomography findings in macular hole closure status after surgery. MATERIALS AND METHODS: Consecutive patients who underwent macular hole surgery were recruited prospectively. All patients underwent detailed macular examination with noncontact and contact biomicroscopy, and linear cross-sectional optical coherence tomography was performed. Macular hole status was defined as elevated/open, flat/open, and flat/closed. Pearson's correlation coefficient was calculated to study the relationship between the clinical and optical coherence tomography findings by assigning a pseudo-value for hole status. RESULTS: Twenty eight patients were recruited, with optical coherence tomography performed successfully in 27 cases. There was good correlation between the optical coherence tomography findings with both the noncontact (r = 0.965; P < .001) and contact lens biomicroscopy (r = 0.965; P < .001). CONCLUSION: Optical coherence tomography findings of postoperative macular hole closure status correlate well with the clinical findings. Careful clinical examination alone may be adequate in determining the surgical anatomical end points in the majority of patients after macular hole surgery. PMID- 12570002 TI - Peripapillary nerve fiber layer thickness measurement reproducibility using optical coherence tomography. AB - OBJECTIVE: To assess reproducibility of quadrantic and clock hour sectors of retinal nerve fiber layer thickness in normal eyes using optical coherence tomography. MATERIALS AND METHODS: Normal eyes of healthy volunteers meeting eligibility criteria were imaged by two inexperienced operators. Six 360 degrees circular scans with a diameter of 3.4 mm centered on the optic disc were obtained during each scanning session, and a baseline image was formed using 3 high quality images defined by the software. Images were obtained on three different days within a 4-week period. Variance and coefficient of variation (CV) were calculated for quadrantic and retinal nerve fiber layer clock hour sectors obtained from the baseline image. RESULTS: Five normal eyes were scanned. Intraoperator reproducibility was high. The mean (+/- SD) CV for total retinal nerve fiber layer thickness was 5.3 +/- 3.82% and 4.33 +/- 3.7% for operators 1 and 2, respectively. Interoperator reproducibility was good with statistically similar variance for all quadrantic and clock hour retinal nerve fiber layer parameters (P = .42 to .99). The nasal retinal nerve fiber layer was the most variable sector for both operators (mean CV: 10.42% and 7.83% for operators 1 and 2, respectively). Differences in mean total, nasal, temporal, and superior retinal nerve fiber layer thickness were not statistically significant between operators for all eyes; however, for inferior retinal nerve fiber layer thickness, there was a significant (P = .0007) difference between operators in one eye. CONCLUSION: Peripapillary retinal nerve fiber layer thickness assessments using optical coherence tomography have good intraoperator and interoperator reproducibility. Inexperienced operators can generate useful measurement data with acceptable levels of variance. PMID- 12570003 TI - Chorioretinal topography and histopathology in laser-induced choroidal neovascularization. AB - OBJECTIVE: To investigate the correspondence between topographic mapping of the vitreoretinal and chorioretinal surfaces in vivo and histopathology findings. MATERIALS AND METHODS: Choroidal neovascularization was induced in the retina of a primate by an argon laser. Serial optical section images of the retina were obtained using an optical imaging system based on the Retinal Thickness Analyzer. Topography of the vitreoretinal and chorioretinal surfaces was mapped. The animal was killed and the eyes enucleated for histopathologic examination. RESULTS: In the normal retina, the topography of the vitreoretinal surface showed a depression at the center of the fovea while the chorioretinal surface was relatively flat, corresponding to normal anatomy. In the retina with choroidal neovascularization, the topography of the vitreoretinal surface indicated a smooth elevation while there were irregular elevations in the topography of the chorioretinal surface. Histological sections displayed focal serous retinal detachment, metaplasia of retinal pigment epithelium, and choroidal neovascularization. CONCLUSION: Topographic mapping of the vitreoretinal and chorioretinal surfaces in vivo corresponds with histological findings and shows promise for quantitative evaluation of pathologic alterations caused by chorioretinal diseases. PMID- 12570004 TI - Subthreshold infrared footprinting with indocyanine green for localizing low intensity infrared photocoagulation. AB - A novel technique for localizing subthreshold infrared (810 nm) laser treatment and its use in three patients is described. Two patients were treated for choroidal neovascularization with transpupillary thermotherapy or focal subthreshold photocoagulation, and the third patient was treated with grid laser photocoagulation for diabetic macular edema using the 810-nm laser after undergoing indocyanine green angiography for diagnostic purposes. Postoperative indocyanine green images, obtained without reinjection of indocyanine green, revealed areas of hypofluorescence that corresponded to the laser treatment spots. This technique, named subthreshold infrared footprinting, can precisely localize subthreshold infrared laser photocoagulation sites and is potentially useful for evaluating treatment postoperatively and planning additional therapy if necessary. PMID- 12570005 TI - Improvements after sheathotomy for branch retinal vein occlusion documented by optical coherence tomography and scanning laser ophthalmoscope. AB - A 55-year-old man presented with decreased visual acuity of 20/400 and central foveal thickness of 450 microm as measured using optical coherence tomography due to a superior temporal branch retinal vein occlusion. Adventitial sheathotomy was performed with a 25-gauge sutureless transconjunctival vitrectomy system. At 1 day, 1 month, and 6 months postoperatively, visual acuity improved to 20/80, 20/40, and 20/30 OS, respectively; central foveal thickness decreased to 228, 195, and 161 microm, respectively; and the scanning laser ophthalmoscope microperimetry showed improved retinal sensitivity and fixation pattern with more stable and central fixation at both postoperative visits. This case indicates optical coherence tomography can detect an early positive effect of sheathotomy surgery on macular edema, and scanning laser ophthalmoscope can document associated improvement in fixation stability. PMID- 12570006 TI - Macular hole secondary to branch retinal vein occlusion diagnosed by Retinal Thickness Analyzer. AB - The Retinal Thickness Analyzer is a laser slit biomicroscopy imaging device that produces accurate measurements of retinal thickness and displays the images in a two-dimensional pattern, superimposed on a fundus image. This article reports a patient with decreased vision following branch retinal vein occlusion in whom the Retinal Thickness Analyzer was used to establish the diagnosis of a macular lamellar hole that was not apparent clinically. PMID- 12570007 TI - Optical coherence tomography of a traumatic Neodymium:YAG laser-induced macular hole. AB - A 34-year-old electronics technician with a macular hole caused by a Neodymium (Nd):YAG laser injury underwent follow up with optical coherence tomography for 1 year. Optical coherence tomography showed a defect in all retinal layers at the macula center, with rectified and perpendicularly orientated borders to the retinal pigment epithelium. One year postinjury, the hole dimensions increased, and optical coherence tomography disclosed less evident rectified borders. After 1 year of follow up, the optical coherence tomography characteristics of a traumatic Nd:YAG laser macular hole can change, looking similar to a senile idiopathic hole. PMID- 12570008 TI - Photodynamic therapy of polypoidal choroidal vasculopathy. AB - Polypoidal choroidal vasculopathy is an abnormality of the choroidal vasculature characterized by aneurysmal-like budding of the vessels with exudation and recurrent serosanguinous detachments of the retinal pigment epithelium. Treatment is limited to laser photocoagulation of leaking vessels, which is used when vision is decreased, or exudate and hemorrhage threaten or involve the fovea. This article describes a patient with polypoidal choroidal vasculopathy successfully treated with photodynamic therapy using verteporfin. PMID- 12570009 TI - Increased diode laser uptake in inner retinal layers after indocyanine green staining of the internal limiting membrane. AB - Intraoperative use of indocyanine green to stain the internal limiting membrane assists in its visualization and removal. This article describes increased uptake of diode laser energy by the superficial layers of the retina after indocyanine green staining of the internal limiting membrane in a patient with diabetic macular edema and a taut, attached posterior hyaloid. The patient was undergoing pars plana vitrectomy, removal of the posterior hyaloid, and indocyanine green assisted peeling of the internal limiting membrane. In addition, panretinal diode laser endophotocoagulation was performed after discovering retinal neovascularization. In areas that were more intensely stained with indocyanine green, the clinical appearance and optical coherence tomography scans demonstrated markedly increased laser energy uptake in the superficial layers of the retina. This case indicates near-infrared or infrared laser procedures performed in areas of indocyanine green-stained internal limiting membrane may necessitate adjustment of laser power and technique. PMID- 12570010 TI - Confocal microscopy in late-onset diffuse lamellar keratitis after laser in situ keratomileusis. AB - This article reports a case of diffuse lamellar keratitis, without exposure of the flap interface, that developed in a patient who underwent intraepithelial photorefractive keratectomy 1 year after bilateral LASIK. Confocal microscopy was performed in both eyes at the onset of the diffuse lamellar keratitis and after its resolution. In the eye with diffuse lamellar keratitis, abundant round structures (inflammatory cells) were present at the interface; these structures disappeared after the keratitis resolved and were not present in the contralateral eye at any time. These confocal microscopic findings further support the hypothesis that diffuse lamellar keratitis is a nonspecific inflammatory response in corneas with a lamellar interface. PMID- 12570011 TI - A new silicone eyecup for ultrasound biomicroscopy. AB - This study compared a new disposable silicone eyecup for high-frequency ultrasound biomicroscopy to the standard hard plastic eyecup. The results showed the silicone eyecup attenuated leakage of the medium in which the probe was immersed, decreased scanning time, decreased distortion of the iris and angle during scanning, enhanced image quality, and improved patient comfort. These findings indicate the silicone eyecup is a good alternative to the hard plastic eyecup and also is easier to use. PMID- 12570012 TI - A wide-field, high-resolution ophthalmoscopic contact lens. AB - A new inverted-image ophthalmoscopic contact lens provides 2X laser spot magnification and a 117 degrees degree instantaneous field of view. It has three times the resolution of very wide-field, inverted-image ophthalmoscopic contact lenses to permit effective use of higher slit-lamp magnifications. The lens was designed for transpupillary thermotherapy with large lesions and panretinal photocoagulation with hazy ocular media. PMID- 12570013 TI - Application of pharmacokinetic modelling to the routine therapeutic drug monitoring of anticancer drugs. AB - Over the last 10 years, proofs of the clinical interest of therapeutic drug monitoring (TDM) of certain anticancer drugs have been established. Numerous studies have shown that TDM is an efficient tool for controlling the toxicity of therapeutic drugs, and a few trials have even demonstrated that it can improve their efficacy. This article critically reviews TDM tools based on pharmacokinetic modelling of anticancer drugs. The administered dose of anticancer drugs is sometimes adjusted individually using either a priori or a posteriori methods. The most frequent clinical application of a priori formulae concerns carboplatin and allows the computation of the first dose based on biometrical and biological data such as weight, age, gender, creatinine clearance and glomerular filtration rate. A posteriori methods use drug plasma concentrations to adjust the subsequent dose(s). Thus, nomograms allowing dose adjustment on the basis of blood concentration are routinely used for 5 fluorouracil given as long continuous infusions. Multilinear regression models have been developed, for example for etoposide, doxorubicin. carboplatin, cyclophosphamide and irinotecan, to predict a single exposure variable [such as area under concentration-time curve (AUC)] from a small number of plasma concentrations obtained at predetermined times after a standard dose. These models can only be applied by using the same dose and schedule as the original study. Bayesian estimation offers more flexibility in blood sampling times and, owing to its precision and to the amount of information provided, is the method of choice for ensuring that a given patient benefits from the desired systemic exposure. Unlike the other a posteriori methods, Bayesian estimation is based on population pharmacokinetic studies and can take into account the effects of different individual factors on the pharmacokinetics of the drug. Bayesian estimators have been used to determine maximum tolerated systemic exposure thresholds (e.g. for topotecan or teniposide) as well as for the routine monitoring of drugs characterized by a very high interindividual pharmacokinetic variability such as methotrexate or carboplatin. The development of these methods has contributed to improving cancer chemotherapy in terms of patient outcome and survival and should be pursued. PMID- 12570014 TI - New insights in insurmountable antagonism. AB - Antagonists that produce parallel rightward shifts of agonist dose-response curves with no alteration of the maximal response are traditionally classified as surmountable, while insurmountable antagonists also depress the maximal response. Although the longevity of the antagonist-receptor complex is quoted in many studies to explain insurmountable antagonism, slowly interconverting receptor conformations, allosteric binding sites, and receptor internalization have been evoked as alternative explanations. To complicate matters even further, insurmountable antagonism is not only drug-related; it may also depend on the tissue, species and experimental design. For the sake of drug development, it is important to elucidate the molecular mechanisms of insurmountable antagonism. New experimental approaches, such as intact cell studies and the use of computer assisted simulations based on dynamic receptor models, herald the advent of better insight in the future. PMID- 12570015 TI - Do endogenous opioids contribute to the bradycardia of rats with obstructive cholestasis? AB - Endogenous opioids have nitric oxide (NO)-dependent cardiovascular actions. In the light of biological evidence of accumulation of endogenous opioids in cholestasis and also existence of NO-dependent bradycardia in cholestatic subjects, this study was carried out to evaluate the role of endogenous opioids in the generation of bradycardia in a rat model of cholestasis. Male Sprague Dawley rats were used to induce cholestasis by surgical ligation of the bile duct, with sham-operated animals serving as a control. The animals were divided into six groups which received naltrexone [20 mg/kg/day, subcutaneously (s.c.)], N(G)-L-nitro-arginine methyl ester (L-NAME, 3 mg/kg/day, s.c.), aminoguanidine (200 mg/kg/day, s.c.), L-arginine (200 mg/kg/day, s.c.), naltrexone + L-NAME (20 and 3 mg/kg/day, s.c) or saline. One week after the operation, a lead II electrocardiogram (ECG) was recorded and the spontaneously beating atria of the animals were then isolated and the chronotropic responses to epinephrine evaluated. The plasma L-nitro-tyrosine level and alanine amino transferase and alkaline phosphatase activities were also measured. The heart rate of cholestatic animals was significantly lower than that of control rats in vivo and this bradycardia was corrected with daily adminstration of naltrexone or L-NAME. The basal spontaneous beating rate of atria in cholestatic animals was not significantly different from that of sham-operated animals in vitro. Cholestasis induced a significant decrease in the chronotropic effect of epinephrine. This effect was corrected by daily injection of naltrexone or L-NAME, or concurrent administration of naltrexone + L-NAME, and was not corrected by aminoguanidine. L arginine had an equivalent effect to L-NAME and increased the chronotropic effect of epinephrine in cholestatic rats but not in control animals. Bile duct ligation increased the plasma activity of liver enzymes as well as the level of L-nitro tyrosine. L-arginine and naltrexone treatment significantly decreased the elevation of liver enzymes in bile duct-ligated rats. Pretreatment of cholestatic animals with naltrexone or L-NAME decreased the plasma L-nitro-tyrosine level. The results suggest that either prevention of NO overproduction or protection against liver damage is responsible for recovery of bradycardia after naltrexone administration. PMID- 12570016 TI - Alpha2-adrenoceptor ligands inhibit alpha1-adrenoceptor-mediated contraction of isolated rat arteries. AB - The experiments in this study were designed to investigate the potential relaxing effects of different compounds known as alpha2-imidazoline ligands (either agonists or antagonists) in isolated rat arteries, and to test the role of nitric oxide (NO) and prostaglandins, in addition to the influence of the nature of the contracting agent in these responses. Segments of mesenteric arteries were isolated and mounted in a small vessel myograph (JP Trading, Aarhus, Denmark) for isometric tension recording, while segments of gracilis muscle arteries were cannulated and studied in the pressurized state using an arteriograph (Living Systems Instrumentation, Burlington, VT, USA). In phenylephrine precontracted mesenteric arteries, the agonists clonidine, BHT920, UK 14304, and rilmenidine, as well as the antagonists idazoxan, yohimbine and rauwolscine, all induced marked relaxations. Similarly, clonidine and idazoxan, both induced marked dilatations of phenylephrine preconstricted gracilis muscle arteries. In both mesenteric and gracilis muscle arteries, the responses to clonidine and idazoxan were not affected by the NO synthase inhibitor (omega)-nitro-L-arginine (L-NA, 10(-5) M) or the cyclooxygenase inhibitor diclofenac (10(-5) M). In mesenteric arteries, the responses to clonidine or idazoxan were similar when the arteries were precontracted by different alpha1-adrenoceptor agonists (phenylephrine, methoxamine or norepinephrine). In contrast, in arteries precontracted by PGF2alpha or endothelin, clonidine induced contractions while idazoxan induced very modest relaxations. Thus, alpha2-adrenoceptor/imidazoline ligands (whether agonists and antagonists) induce paradoxical relaxation of small mesenteric or gracilis muscle arteries of rats, which are not affected by NO-synthase or cyclooxygenase inhibition, and appear related to direct non specific interactions of the alpha2-imidazoline ligands with alpha1-adrenergic receptors in vascular smooth muscle. PMID- 12570017 TI - Wine polyphenols modulate calcium handling in rat aorta: involvement of nitric oxide pathway. AB - The effects of short-term oral administration of red wine polyphenolic compounds (RWPCs) on blood pressure and vascular reactivity were investigated in rats. The consequence of RWPCs treatment on agonist-induced contractility of rat aorta with respect to Ca2+ handling was assessed, by examining both intracellular Ca2+ store and extracellular Ca2+ influx components of the response. Rats were treated daily for 7 days by intragastric administration of either 5% glucose, or RWPCs (20 mg/kg) [from two different sources, i.e. Provinols (SFD, Vallont Pont d'Arc, France) and RWPC1 (INRA, Montpellier, France)]. Administration of these compounds produced a decrease in systolic blood pressure. The consequence of RWPCs treatment on vascular smooth muscle was investigated in rat aorta without endothelium exposed to noradrenaline. In Ca(2+)-free medium, RWPC1 but not Provinols treatment induced an increase in noradrenaline-induced contraction. After depletion of intracellular Ca2+ stores by noradrenaline in Ca(2+)-free medium, addition of CaCl2 in the continuous presence of agonist induced an increase in contraction, which was not significantly different between control, Provinols- and RWPC-treated rats. The presence of an inhibitor of sarcoplasmic reticulum Ca(2+)-ATPase, thapsigargin, significantly reduced noradrenaline induced contraction in Ca(2+)-free medium in RWPCs-treated aorta, as compared to that of control. Interestingly, the inhibitory effect of thapsigargin on the response linked to the release of Ca2+ from internal stores in RWPCs-treated vessels was completely prevented in the presence of NO-synthase inhibitor, L nitro arginine methyl ester, the inhibitor of guanylyl cyclase, oxadiazolo quinoxaline or the protein kinase G inhibitor, 8-Bromoguanosine-3'-5-cyclic mono phosphorothioate, Rp isomer. These results suggest that short-term administration of RWPCs in rats induced subtle alteration of thapsigargin-sensitive component of agonist-induced contraction in rat aorta linked to Ca2+ release from intracellular store. Calcium release from intracellular stores sensitive to thapsigargin was implicated in this mechanism. The prevention of the inhibitory effect of thapsigargin by the inhibitors of NO/cyclic guanosine monophosphate pathway after RWPCs treatment highlights the role of NO in this phenomenon. PMID- 12570018 TI - Effect on cancer cell proliferation of palmitoylethanolamide, a fatty acid amide interacting with both the cannabinoid and vanilloid signalling systems. AB - Palmitoylethanolamide (PEA) is a bioactive fatty acid amide belonging to the class of N-acyl-ethanolamines (NAEs). This compound has been known since the 1950s for its anti-inflammatory effects, but was re-discovered only after the finding that another NAE, arachidonoyl-ethanolamide (anandamide, AEA), could act as an endogenous ligand of cannabinoid receptors. Although a similar function for PEA has also been proposed, this compound does not activate the two cannabinoid receptor subtypes described to date. PEA and AEA are co-synthesized by cells, and PEA might act as an 'entourage' compound for AEA, i.e. as an endogenous enhancer of AEA biological actions. Indeed, long-term treatment of human breast cancer cells (HBCCs) with PEA downregulates the expression of the enzyme responsible for AEA degradation, the fatty acid amide hydrolase, thereby leading to an enhancement of AEA-induced, and cannabinoid CB1 receptor-mediated, cytostatic effect on HBCCs. AEA is also a full agonist for the receptors of another class of bioactive fatty acid amides, the N-acyl-vanillyl-amines (e.g. capsaicin and olvanil). These sites of action are known as vanilloid receptors of type 1 (VR1). PEA enhances the VR1-mediated effects of AEA and capsaicin on calcium influx into cells. These 'entourage' effects of PEA might be attributable to modulation of VR1 activity, and could underlie the enhancement by PEA, described here for the first time, of the antiproliferative effects of VR1 receptor agonists. PMID- 12570019 TI - Nimesulide and diclofenac inhibit lipopolysaccharide-induced hypothermia and tumour necrosis factor-alpha elevation in rats. AB - The effects of nimesulide and diclofenac on lipopolysaccharide (LPS)-induced rectal temperature changes and serum tumour necrosis factor (TNF)-alpha elevation were investigated in rats. LPS (Escherichia coli O111:B4; 50 microg/kg, intraperitoneally) produces a dual body temperature response, in which initial hypothermia precedes fever. Serum TNF-alpha levels rise during the initial phase of the induced hypothermia. Nimesulide, a preferential inhibitor of cyclooxygenase-2 (0.05, 0.5 or 1 mg/kg, subcutaneously) completely abolished the hypothermia, resulting in an acceleration of the fever phase. However, the peak and plateau phases of fever were not changed by nimesulide treatment. Nimesulide (0.5 mg/kg) partially prevented serum TNF-alpha elevation. The non-selective cyclooxygenase inhibitor diclofenac inhibited hypothermia at all doses tested (0.03, 0.3 or 3 mg/kg, subcutaneously) although fever was completely abolished at the 3 mg/kg dose only. Diclofenac also partially abolished the elevation in serum TNF-alpha levels, but at the highest dose only (3 mg/kg). These data suggest that nimesulide and diclofenac can preferentially inhibit LPS-induced hypothermia at doses that do not abolish fever in rats. Both these drugs also reduced elevated TNF-alpha levels, a fact which may, at least partly, explain the antihypothermic effect of nimesulide. PMID- 12570020 TI - Effect of amitriptyline on gastric ulceration. AB - Amitriptyline significantly inhibited alcohol, aspirin, indomethacin and cold restraint stress-induced ulceration. Secretory studies conducted in pyloric ligated rats revealed that the drug, at the doses employed, significantly reduced total acidity and protein content. However, significant reductions of the gastric volume were only observed at the highest dose of the drug. In another set of experiments, when 50% alcohol (v/v) was administered to the pyloric-ligated rats pretreated with amitriptyline, it was observed that the drug significantly reduced the pH, total acidity and protein content. PMID- 12570022 TI - Membrane effects of ropivacaine compared with those of bupivacaine and mepivacaine. AB - We compared the effects of ropivacaine, bupivacaine and mepivacaine on membrane lipids in an attempt to determine the anaesthetic mechanism of ropivacaine with structure-dependent potency. The membrane effects were determined by measuring anaesthetic-induced changes in the phase transition temperature and the fluorescence polarization of liposomal membranes prepared with cholesterol and phosphatidylcholine. Bupivacaine, ropivacaine and mepivacaine depressed the membrane lipid phase transition and decreased the polarization of liposomal membranes at 0.0625-1.0 mg/mL, indicating that these anaesthetics fluidize membranes at concentrations lower than those in clinical use. Ropivacaine and bupivacaine were effective in fluidizing the membrane core rather than the membrane surface, whereas mepivacaine was a membrane fluidizer acting equally on both regions. In the comparison of membrane fluidization at an equimolar concentration (3.0 mmol/L), ropivacaine was found to be less potent than bupivacaine and more potent than mepivacaine. This membrane-fluidizing potency was also consistent with the hydrophobic properties of these substances evaluated by reversed-phase chromatography. Structure-dependent membrane fluidization associating with hydrophobicity appears to underlie the local anaesthetic effect of ropivacaine as well as those of bupivacaine and mepivacaine. PMID- 12570021 TI - Binding of KRH-594, an antagonist of the angiotensin II type 1 receptor, to cloned human and rat angiotensin II receptors. AB - We studied the binding properties of KRH-594, a new selective antagonist of angiotensin II (AII) type 1 (AT1) receptors, to rat liver membranes and to recombinant AT1 and AT2 receptors. Preincubation of rat liver membranes with KRH 594 produced maximal inhibition of [125I]-AII binding when the preincubation time was 1-2 h. Preincubation with KRH-594 for 2 h decreased the B(max) value and increased the Kd value. For human AT1, human AT2, rat AT1A and rat AT1B receptors, the Ki values for KRH-594 were 1.24, 9360, 0.67, and 1.02 nm, respectively. The rank order of K1 values for human AT1 receptors was KRH-594 >> EXP3174 > candesartan = AII. The order of specificities for human AT1 and AT2 receptors was candesartan > EXP3174 > KRH-594. Although a 2-h preincubation of human AT2 receptors with KRH-594 (30 microM) or CGP 42112 (a selective AT2 receptor antagonist; 0.3 nM) inhibited binding of [125I]-AII, the suppression by KRH-594 was not significant. These results indicate that KRH-594 binds potently to AT1 receptors in an insurmountable manner, and that at a very high dose (30 microM) it may also bind to AT2 receptors, but in a surmountable manner. PMID- 12570023 TI - Proton pump inhibitors in the elderly population. AB - Accurate diagnosis and effective treatment of acid-related diseases, including gastro-oesophageal reflux disease (GORD), are important health care priorities, particularly in the elderly. Both the prevalence and severity of GORD are increased in older individuals. The reason for the age-related increase in the prevalence of GORD is not completely understood, but it appears to result from both age-related changes in physiology and effects of the medications often taken by older people. The diagnosis of GORD in the elderly is also difficult because of its potential atypical presentation in these patients, as well as the overlap between GORD symptoms and those of other chronic conditions, including coronary artery disease and chronic obstructive pulmonary disease. Proton pump inhibitors (PPIs) are now considered the treatment of choice for patients with GORD, and these drugs have excellent efficacy and safety profiles. One of the newer drugs in this class, rabeprazole, may be particularly suited for use in older patients. Rabeprazole requires no dosing adjustment in the elderly or in patients with renal insufficiency or mild-to-moderate hepatic disease. It is also relatively free of clinically significant drug-drug interactions, which strongly differentiates rabeprazole from omeprazole, the prototype PPI. Overall, the clinical and pharmacokinetic profiles for rabeprazole suggest that it is an excellent first choice for the management of older patients with GORD. PMID- 12570024 TI - Gastrointestinal disease and the kidney. AB - Renal disease at any stage, from insufficiency to end-stage renal disease requiring dialysis or following renal transplantation, is often accompanied by significant gastrointestinal (GI) symptoms. Conversely, patients with GI disease may present with significant renal complications. Patients with end-stage renal disease, patients undergoing dialysis, and recipients of renal grafts are at increased risk for GI complications, including erosive disease and GI bleeding. Selection of pharmacotherapy for GI conditions in patients with concomitant renal disease is complicated by three factors: (1) the potential for a significant negative impact on renal function, which is already compromised; (2) the requirement for dosing alteration in renal insufficiency; and (3) the potential for drug-drug interactions with concomitant medications. Proton pump inhibitors appear to be the most suitable acid-suppressing therapy for patients with renal disease; recently developed drugs in this class (e.g. rabeprazole) may be the best choice for treatment of patients with both acid-related GI conditions and renal disease. PMID- 12570025 TI - Gastro-oesophageal complications in patients receiving cancer therapy: the role of proton pump inhibitors. AB - Chemotherapy and radiotherapy, both alone and in combination, remain important tools in the successful treatment of cancer. Because the oral and gastrointestinal (GI) mucosa is often significantly damaged by cancer therapy, management of these problems is an important challenge for oncologists. Such treatment complications are generally not severe or life threatening, but they can result in both treatment delays and dose reductions in potentially curative regimens. Numerous therapeutic approaches have been evaluated as prophylaxis or treatment for mucosal damage in patients undergoing cancer therapy. The results of large-scale, placebo-controlled, comparative trials demonstrate that administration of a proton pump inhibitor (PPI) can provide both significant symptom relief and prophylaxis against upper GI ulceration in patients receiving cancer chemotherapy. Although no corresponding clinical trial has been conducted in patients undergoing irradiation, PPIs are also likely to be effective in preventing gastro-oesophageal mucosal injury in such individuals. Thus, PPIs may play a crucial role in supportive care for patients undergoing cancer therapy. PMID- 12570026 TI - Laryngopharyngeal manifestations of reflux: diagnosis and therapy. AB - Acid reflux--most often associated with heartburn--may also cause a wide range of laryngopharyngeal symptoms, including laryngitis and chronic cough. Symptoms of laryngopharyngeal reflux (LPR), like those of gastrooesophageal reflux disease, result from abnormal exposure of tissues to acid refluxate. Deranged sensorimotor function of the upper oesophageal sphincter appears to play a key role in the aetiology of LPR, but the disease is not completely understood. Among the significant long-term complications of LPR are bronchopulmonary disorders, recurrent pneumonia, chronic cough, chronic or recurrent laryngitis, and oral cavity disorders. It also appears to be a risk factor for the development of laryngeal carcinoma. Diagnosis of LPR is based on physical examination, medical history, and results of specific tests. At present, the test of choice for LPR diagnosis is intraluminal oesophageal pH monitoring. Barium contrast oesophagography, intraoesophageal acid perfusion challenge, and flexible endoscopic evaluation of swallowing with sensory testing may also be used in LPR diagnosis. Treatment for LPR includes changes to the diet and lifestyle, and acid suppressing therapy. The Therapeutic Working Party at the First Multi Disciplinary International Symposium on Supraesophageal Complications of Gastroesophageal Reflux Disease has recommended twice-daily dosing with a proton pump inhibitor as an initial therapy for LPR, with treatment continued between 4 weeks and 6 months. Such treatment has been found highly effective in resolving symptoms of LPR, and it may also prevent the serious long-term complications of this condition. PMID- 12570027 TI - Do selective cyclo-oxygenase inhibitors eliminate the adverse events associated with nonsteroidal anti-inflammatory drug therapy? AB - Among the most widely prescribed drugs worldwide, non-steroidal anti-inflammatory drugs (NSAIDs) are effective for relieving pain, but they are also associated with a high incidence of gastrointestinal (GI) adverse events. Both the beneficial and harmful effects of NSAIDs result from inhibition of the cyclo oxygenase (COX) enzyme. Recognition of the two distinct COX isoforms prompted development of drugs that selectively block the activity of COX-2, thus providing pain relief and reducing inflammation while sparing COX-1, the enzyme apparently responsible for most protective prostaglandin synthesis in the mucosa of the stomach and duodenum. The results of preclinical and clinical studies indicate that COX-2 inhibitors exhibit high selectivity in inhibiting COX-2, provide excellent pain relief, and cause significantly less GI toxicity than do conventional nonselective NSAIDs. Although they represent a significant advance over nonselective NSAIDs, selective COX-2 inhibitors are not without limitations. They do not completely eliminate GI toxicity or the renal side effects associated with use of conventional NSAIDs. Moreover, in cases of inflammation or ulceration in the GI tract, COX-2 inhibition may delay ulcer healing. Finally, case reports and the results of animal experiments suggest that COX-2 inhibitors may adversely affect ovulation and cause a tendency towards prothrombotic events. PMID- 12570028 TI - In vitro tests for predicting drug-drug interactions: the need for validated procedures. AB - Over the past decade, the prediction of drug-drug interactions from in vitro studies has become a rapidly expanding field of research. Numerous papers and excellent review articles (Bertz & Granneman 1997; Ito et al. 1998a & b; Lin 2000; Bachmann & Ghosh 2001; Ekins & Wrighton 2001; Weaver 2001) have been published in this area. Yet like any new and fast-growing subject, this one has been developing with some confusion and without any real, efficient organisation. Depending on the drug tested, the models and extrapolation parameters used, etc., results and conclusions may vary widely from study to study (von Moltke et al. 1998; Weaver 2001). Several authors have called for validation of these procedures (Rodrigues et al. 2001; Kummar & Surapaneni 2001; Pelkonen et al. 2001a & b; Kremers 2002), and regulatory authorities intend to require better traceability and reliability (FDA & EMEA guidelines). A systematic and reliable approach is needed also to allow such protocols to be incorporated into early screening for potential drugs and new chemical entities. There is certainly a great need to standardise these studies and to verify their conclusions, but is true validation possible in this field? The main purpose of the present paper is to discuss this issue and to examine what is possible and what is needed to improve the quality of predictions made from in vitro experiments. PMID- 12570029 TI - Subchronic toxicity of Baltic herring oil and its fractions in the rat I: Fractionation and levels of organohalogen pollutants. AB - Baltic herring (Clupea harengus) oil was extracted and fractionated. To examine the contribution to toxicity and biological effects of different halogenated organic pollutants, the herring oil and the fractions were mixed into pelleted food and given to Sprague-Dawley female rats at three levels, corresponding to a human intake of 1.6, 8.2 and 34.4 kg fish per week. Herring oil, its fractions, as well as liver tissues from exposed rats, were analyzed for: eight chlorinated biphenyls, all 2,3,7,8-substituted chlorinated dibenzo-p-dioxins and dibenzofurans, hexachlorocyclohexanes, hexachlorobenzene, 1,1,1-trichloro-2,2 bis(4-chlorophenyl)ethane (DDT), DDT-metabolites, three brominated diphenylethers as well as extractable organically bound chlorine and halogenated fatty acids. A bioassay (EROD) was used for measuring the dioxin-like enzyme induction activity. Nordic Sea lodda (Mallotus villosus) oil was used as a nutritionally equivalent control, with much lower levels of halogenated organic pollutants. A full toxicological subchronic examination is reported in the following paper (Stern et al. 2002). In this study, we report that the fractionation procedure resulted in a substantial reduction of most of the pollutants in the triacylglycerol fraction, and a pronounced enrichment of most of the pollutants into the two other fractions. However, all contaminants were present at some levels in all of the fractions. The concentrations of organohalogens found in this study were representative for Baltic herring during the mid-1990s. Rat liver tissue showed similar residue patterns as the diet, with the exception of chlorinated dibenzo-p dioxin and dibenzofuran congeners that had a higher liver retention than pesticides, chlorinated biphenyls and brominated diphenylethers. PMID- 12570030 TI - Subchronic toxicity of Baltic herring oil and its fractions in the rat II: Clinical observations and toxicological parameters. AB - This study aimed to increase the knowledge about the toxicity of fish-derived organohalogen pollutants in mammals. The strategy chosen was to separate organohalogen pollutants derived from Baltic herring (Clupea harengus) fillet, in order to obtain fractions with differing proportions of identified and unidentified halogenated pollutants, and to perform a subchronic toxicity study in rats, essentially according to the OECD guidelines, at three dose levels. Nordic Sea lodda (Mallotus villosus) oil, with low levels of persistent organohalogen pollutants, was used as an additional control diet. The toxicological examination showed that exposure to Baltic herring oil and its fractions at dose levels corresponding to a human intake in the range of 1.6 to 34.4 kg Baltic herring per week resulted in minimal effects. The spectrum of effects was similar to that, which is observed after low-level exposure to pollutants such as chlorinated dibenzo-p-dioxins and dibenzofurans (CDD/F) and chlorinated biphenyls, despite the fact that these contaminants contribute to a minor part of the extractable organically bound chlorine (EOCI). The study confirmed previous findings that induction of hepatic ethoxyresorufin deethylase (EROD) activity takes place at daily intake levels 0.15 ng fish-derived CDD/F TEQs/kg body weight. The study also demonstrated that hepatic vitamin A reduction takes place at somewhat higher daily exposure levels, i.e. 0.16-0.30 ng fish derived CDD/F-TEQs/kg body weight. Halogenated fatty acids, the major component of EOCI, could not be linked to any of the measured effects. From a risk management point of view, the study provides important new information of effect levels for Ah-receptor mediated responses following low level exposure to organohalogen compounds from a matrix relevant for human exposure. PMID- 12570031 TI - In vitro cytotoxicity of protocatechuic acid to cultured human cells from oral tissue: involvement in oxidative stress. AB - Data on the biologic activity of protocatechuic acid are contradictory; some studies have shown that it acts as an antioxidant and suppresses chemical-induced carcinogenesis and others that it induces oxidative stress and promotes tumour formation. The anticarcinogenicity of protocatechuic acid was postulated to be related, in part, to its specific suppression of neoplastic hyperproliferation. To determine whether protocatechuic acid was preferentially antiproliferative to malignant cells, non-malignant and carcinoma cells were exposed for 24 hr to protocatechuic acid (2.5 to 25 mM) and viability was assessed with the neutral red assay. The cell lines were derived from tissues of the human oral cavity, the initial site of exposure upon ingestion of dietary protocatechuic acid, and included normal GN61 gingival fibroblasts, immortalized, non-tumorigenic S-G gingival epithelial cells, and malignant HSG1 cells derived from the salivary gland, HSC-2 cells from the floor of the oral cavity, and CAL27 cells from the tongue. Selective toxicity of protocatechuic acid to malignant cells was not observed. Furthermore, using a total cellular protein determination to quantitate cell growth, no differences in comparative sensitivities of S-G epithelial cells and HSG1 carcinoma cells were noted in a 3 day continuous exposure to 2.5 to 12.5 mM protocatechuic acid and in recovery from a 24 hr exposure to 3 to 15 mM protocatechuic acid. The S-G and HSG1 cells were then used to study the effects of elevated concentrations of protocatechuic acid on oxidative stress. For both cell types, protocatechuic acid induced oxidative stress, presumably through its bioactivation by a tyrosinase pathway. A brief exposure to 25 mM protocatechuic acid lowered the levels of intracellular glutathione and potentiated Fe2+-induced lipid peroxidation of the cells. As determined with the neutral red assay, S-G and HSG1 cells exposed briefly to a non-toxic level (0.5 mM) of the glutathione depleter, 1,3-bis(2-chloroethyl)-N-nitrosourea, were hypersensitive to a subsequent challenge with 10 mM protocatechuic acid and preexposure of the S-G and HSG1 cells to a nontoxic level of protocatechuic acid (2.5 mM) enhanced their sensitivity to a subsequent exposure to tert-butyl hydroperoxide. These findings were consistent with protocatechuic acid, at high levels (> or = 10 mM), acting as an inducer of oxidative stress. PMID- 12570032 TI - The in vitro effects of galactose and its derivatives on rat brain Mg2+-ATPase activity. AB - Galactosaemia is an inborn error of metabolism characterized by irreversible damage to neural tissue. To evaluate whether galactose metabolic disorders, (e.g. classical galactosaemia, galactokinase deficiency galactosaemia), is implicated for alterations of brain Mg2+-ATPase activity, various concentrations (1-16 mM) of galactose, galactose-1-phosphate, galactitol, glucose-1-phosphate or glucose were preincubated with whole brain homogenates of suckling rats at 37 degrees for 1 hr. Mg2+-ATPase activities were determined according to Bowler & Tirri's (1974). Galactose-1-phosphate or glucose-1-phosphate excessively activated the brain Mg2+-ATPase in a concentration-dependent way. Additionally, galactitol, galactose or glucose stimulated the enzyme up to 35-45% (P < 0.001) at concentrations >4 mM. A mixture of galactose-1-phosphate (2 mM), glactitol (2 mM) and galactose (4 mM), concentrations commonly found in blood and brain of untreated patients with classical galactosaemia, resulted in a 500% enzyme activation (P < 0.001) as compared to control. Moreover, a mixture of galactitol (2 mM) and galactose (1 mM), concentrations measured in patients with galactokinase deficiency, caused an enzyme stimulation (35%, P < 0.001). These findings suggest: a) The great Mg2+-ATPase activation by galactose-1-phosphate or glucose-1-phosphate may be due to the epimer of galactose and the presence of phosphorus. b) The brain Mg2+-ATPase stimulation by galactose and its derivatives could be toxic by modulating the Mg2+ concentration, the ATP availability, the activity of other ATP- and Mg2+-dependent enzymes as well as the rates of protein synthesis and cell growth. PMID- 12570033 TI - Diazenecarboxamide UP-91, a potential anticancer agent, acts by reducing cellular glutathione content. AB - Glutathione (GSH) is a ubiquitous non-protein thiol essential for cellular homeostasis and protection. Diazenecarboxamides (diazenes) are new compounds that could, according to their biochemical properties, lower the intracellular GSH content, thus inhibiting the growth of tumour cells. In the present study we examined four such compounds: JK-914, JK-918, JK-1013 and UP-91. Their cytotoxic effect on the growth of eight human tumour cell lines (glioblastoma, cervical and laryngeal carcinoma cells, mammary carcinoma cells and four drug-resistant sublines) was determined using a modified colorimetric MTT assay. The rate of reaction of thiophenol (as a model thiol) with diazenes leading to diphenyl disulfide was established by chromatography (TLC). Reactivity of diazenes with GSH under quasi-physiological conditions was determined by NMR spectroscopy. Intracellular GSH content was examined spectrophotometrically by the procedure developed by Tietze (1969). Diazene UP-91 reduced significantly the cell survival of all eight examined cell lines, including four drug-resistant cell lines. Other diazenes did not influence the survival of tumour cells. Reaction time for quantitative conversion of thiophenol to diphenyl disulfide was shortest for diazene UP-91, which is highly consistent with high reactivity of the same diazene with GSH, observed under quasi-physiological conditions. UP-91 reduced intracellular GSH level, while other diazenes had no effect on it. Thus, diazenecarboxamides UP-91 is a potential anticancer agent that may inhibit the growth of tumour cells due to reduction in glutathione level. PMID- 12570034 TI - Adjuvant effect of di-n-butyl-, di-n-octyl-, di-iso-nonyl- and di-iso-decyl phthalate in a subcutaneous injection model using BALB/c mice. AB - During the last decades, the prevalence of the allergic airway diseases, asthma and rhinitis, has increased world-wide. Introduction of environmental chemicals with adjuvant effect may play a role in this increase. In the present study, the adjuvant effects of di-n-butyl-, di-n-octyl-, di-iso-nonyl- and di-iso-decyl phthalate are studied in a screening model. Ovalbumin, used as the model antigen, was injected subcutaneously in the neck region of BALB/cJ mice with the selected phthalate in concentrations from 2-2000 microg/ml. Additionally, the mice were boosted once or twice with ovalbumin alone. Immunization with ovalbumin alone, the ovalbumin control group, served as the baseline for antibody production, whereas aluminium hydroxide served as the positive control. The levels of ovalbumin-specific IgE, IgG1 and IgG2a antibodies in sera were determined. Adjuvant effect was accepted to be present if a statistical increase in antibody production occurred in a test group as compared to an ovalbumin control group together with the fulfillment of dose-response relationships. Adjuvant effect varied strongly between the phthalates investigated. Phthalates with 8 or 9 carbon atoms in the alkyl side chains were the stronger adjuvants whereas phthalates with shorter or longer alkyl side chains possessed less adjuvant activity. Adjuvant effects were apparent either from the IgE or the IgG1 response or both, whereas no effect was seen on the IgG2a response. Additional studies with airborne exposure are required to establish whether the hazards also result in a significant risk for the development of allergy in man. PMID- 12570035 TI - Toward optimal health: the experts discuss cognitive function and memory changes. PMID- 12570036 TI - Bony pathology in the cancer patient. AB - Bony pathology in the cancer patient represents a significant source of morbidity and mortality. Complications include insufficiency and pathological fractures resulting from either medical treatments or bony metastases that can cause significant functional limitations. Additional complications include spinal cord compression, hypercalcemia, and bone marrow failure. Rehabilitation management of such conditions is reviewed, with an emphasis on diagnostic and therapeutic management. Bracing and focused rehabilitation programs facilitate maximal participation and functional outcomes, which can result in an enhanced quality of life. Specific rehabilitation goals and strategies are discussed, with an emphasis on tailoring these according to the functional staging of the patient. PMID- 12570037 TI - Quality of life and menopause: the role of estrogen. AB - The use of estrogen or hormone replacement therapy (ERT/HRT) in preventing disease in menopausal women has been well documented. Less attention has been paid to the menopausal symptoms that can impair the quality of life of menopausal women, such as hot flushes, sleep disorders, sexual dysfunction, and alterations in mood. Researchers have used a variety of methods to investigate these concerns. Decreases in ovarian hormones that occur with menopause have been implicated in these symptoms. Ovarian hormones affect the central nervous system and urogenital tissues directly via receptors for estrogen, progesterone, and androgens. Changes in the symptoms of menopause consequential to estrogen therapy reflect the effect of this therapy on these tissues. Evidence supporting the effectiveness of ERT/HRT in the treatment of symptoms affecting quality of life is growing and supports the use of ERT/HRT during menopause. Because the most dramatic hormonal changes associated with menopause are related to estrogen and because estrogen is usually coadministered with a progestogen in patients with an intact uterus, this review is focused primarily on ERT/HRT. Because androgen therapy may also improve quality of life by enhancing perimenopausal and postmenopausal sexual desire, function, and general well-being, a brief discussion of androgen supplementation of ERT/HRT is also included. The ideal doses and combinations of hormones must be determined on an individual basis, taking into consideration benefits, risks, and interactions of the different hormone therapies. PMID- 12570038 TI - Chest pain in hospitalized patients: cause-specific and gender-specific differences. AB - OBJECTIVES: The aim of this prospective cardiological-linguistic study was to assess cause-specific and gender-specific differences in the reported symptoms and description of chest pain. METHODS: In patients hospitalized because of chest pain, location, radiation, quality of chest pain, pain precipitating and relieving factors, and additional symptoms were assessed. The cause of chest pain was assessed as either coronary or noncoronary. Patients' pain descriptions were taped for linguistic narrative analysis and transcribed according to ethnomethodological standards. RESULTS: The cause of chest pain was assessed as coronary in 43 (18 females, 25 males, mean age 63 years) and noncoronary in 49 (30 females, 19 males, mean age 62 years) patients. Only few cause-related differences in the symptoms were found. In patients with a coronary cause, the location of chest pain was more often retrosternal (93% vs. 71%, p = 0.0078), in the right arm (23% vs. 6%, p = 0.0186), and less often in the back (28% vs. 51%, p = 0.0241) than in patients with a noncoronary cause of chest pain. Coronary patients more often had a pressing pain quality (81% vs. 61%, p = 0.034), less often pain precipitated by respiration (16% vs. 45%, p = 0.0032), and vertigo (21% vs. 43%, p = 0.0252) than noncoronary patients. The women were older than the men (mean age 65.6 vs. 59.0 years, p = 0.01). Women with a coronary cause more often had a gradual pain onset (78% vs. 48%, p = 0.0488) and relief by rest (78% vs. 40%, p = 0.0139) than men with a coronary cause. Linguistic analysis revealed that men presented themselves as interested in the cause of the chest pain, observing and describing pain concretely, whereas women presented themselves as prevailingly pain enduring, describing their pain diffusely. CONCLUSIONS: Cause-related and gender-related differences in symptoms are too unspecific to distinguish between coronary and noncoronary causes. The strong gender differences in self-presentation and description of chest pain might be an explanation for underdiagnosis and undertreatment of women with coronary heart disease and should be considered when taking the clinical history of a female patient. PMID- 12570039 TI - Deaf women: experiences and perceptions of healthcare system access. AB - BACKGROUND: The authors investigated the knowledge, attitudes, and healthcare experiences of Deaf women. METHODS: Interviews with 45 deaf women who participated in focus groups in American Sign Language were translated, transcribed, and analyzed. Deaf women's understanding of women's health issues, knowledge of health vocabulary in both English and American Sign Language, common health concerns among Deaf women, and issues of access to information, including pathways and barriers, were examined. As a qualitative study, the results of this investigation are limited and should be viewed as exploratory. RESULTS: A lack of health knowledge was evident, including little understanding of the meaning or value of cancer screening, mammography, or Pap smears; purposes of prescribed medications, such as hormone replacement therapy (HRT); or necessity for other medical or surgical interventions. Negative experiences and avoidance or nonuse of health services were reported, largely due to the lack of a common language with healthcare providers. Insensitive behaviors were also described. Positive experiences and increased access to health information were reported with practitioners who used qualified interpreters. Providers who demonstrated minimal signing skills, a willingness to use paper and pen, and sensitivity to improving communication were appreciated. CONCLUSIONS: Deaf women have unique cultural and linguistic issues that affect healthcare experiences. Improved access to health information may be achieved with specialized resource materials, improved prevention and targeted intervention strategies, and self-advocacy skills development. Healthcare providers must be trained to become more effective communicators with Deaf patients and to use qualified interpreters to assure access to healthcare for Deaf women. PMID- 12570040 TI - Continuity of medical care and risk of incarceration in HIV-positive and high risk HIV-negative women. AB - OBJECTIVES: Incarceration rates in the United States have tripled over the past two decades and have increased even more rapidly among women than men. To identify risk factors that predict incarceration in HIV-positive (HIV+) and high risk HIV-negative (HIV-) women and to evaluate the association between continuity of medical care and risk of incarceration, this analysis was conducted. METHODS: This was a prospective cohort study of HIV+ and high-risk HIV- women enrolled between April 1993 and January 1995 at four urban centers: Providence, Rhode Island; New York, New York; Baltimore, Maryland; and Detroit, Michigan. The HIV Epidemiology Research (HER) Study enrolled 871 HIV+ and 439 high-risk HIV- innercity women between the ages of 16 and 55 years. All participants had a history of injection drug use or high-risk sexual behavior. Interviews, including questions on continuity of medical care and incarceration, were administered at baseline and 6 and 12 months after enrollment. Any incarceration in the 1-year period following enrollment was the main outcome measure. Continuity of care was measured as having seen one healthcare provider for at least 2 years, having received medical care from one usual physician or clinic, and having obtained medical care in a primary care setting as opposed to an emergency room or drug treatment center. RESULTS: Twelve percent of women were incarcerated within 1 year postenrollment. Factors associated with incarceration included recent drug use, prior incarceration, lack of college education, engaging in sex for drugs or money, and having multiple unmet basic needs at the time of enrollment in the study. Continuity of care with a single healthcare provider for more than 2 years prior to enrollment in the study was associated with decreased rates of incarceration even after adjusting for possible confounding factors (OR = 0.67, 95% CI = 0.48 - 0.92). HIV serostatus did not correlate with incarceration. CONCLUSIONS: History of prior incarceration and recent drug use were associated with increased risk of incarceration. Continuity of medical care by a single healthcare provider was associated with decreased likelihood of incarceration, suggesting that the provider may play an important role in designing interventions to prevent incarceration in this high-risk population. PMID- 12570041 TI - Implication of SAR of male medfly attractants in insect olfaction. AB - Medfly (Ceratitis capitata) males are strongly attracted by different compounds, not described as pheromones. The best attractants reported are (+)-alpha-copaene, a sesquiterpene of natural source and (-)-ceralure-B1, a non-natural iodinated cyclohexane ester. Although their origin, atomic composition, chemical and physical properties are rather different, they show similar attraction to medflies. The question of why these compounds, act behaviorally in the same way, has been never addressed in research papers. We show here for the first time that these compounds have quite similar stereochemistry, water accessible surfaces, certain local dipole moments and, to some extent, similar octanol/water partition coefficient (log P). When seven carbons, one oxygen and one iodine belonging to ( )-ceralure-B1 are selectively chosen based on topological homology with (+)-alpha copaene and are overlaid with nine corresponding carbons of (+)-alpha-copaene, the RMS is 0.367 A. This represents a high degree of steric resemblance. Local dipole moments and charges are similar in those regions where the molecules show topological homologies. Thus, we hypothesize that these two molecules could interact with the same male medfly's odorant receptor(s). The implications of this result in future research in insect olfaction is discussed. PMID- 12570042 TI - Structure-toxicity modeling of pesticides to honey bees. AB - A quantitative structure-activity relationship (QSAR) model was derived for estimating the acute toxicity of pesticides on the honey bee. Chemicals were described by means of autocorrelation descriptors encoding lipophilicity (H), molar refractivity (MR) and the H-bonding acceptor ability (HBA) of the pesticides. A three-layer feedforward neural network trained by the back propagation algorithm was used as statistical engine for deriving a powerful QSAR model. The root mean square residual (RMSR) values for the training and testing sets were 0.430 and 0.386, respectively. The practical interest of this original model was discussed. PMID- 12570043 TI - Prediction of tissue-air partition coefficients: a comparison of structure-based and property-based methods. AB - Three linear regression methods were used to develop models for the prediction of rat tissue-air partition coefficient (P). In general, ridge regression (RR) was found to be superior to principal component regression (PCR) and partial least squares regression (PLS). A set of 46 diverse low molecular-weight volatile chemicals was used to model fat-air, liver-air and muscle-air partition coefficients for male Fischer 344 rats. Comparisons were made between models developed using descriptors based solely on molecular structure and those developed using experimental properties, including saline-air and olive oil-air partition coefficients, as independent variables, indicating that the structure property correlations are comparable to the property-property correlations. Multiple structure-based models were developed utilizing various classes of structural descriptors based on level of complexity, i.e. topostructural (TS), topochemical (TC), 3-dimensional (3D) and calculated octanol-water partition coefficient. In most cases, the structure-based models developed using only the TC descriptors were found to be superior to those developed using other structural descriptor classes. Haloalkane subgroups were modeled separately for comparative purposes, and although models based on the congeneric compounds were superior, the models developed on the complete sets of diverse compounds were acceptable. Comparisons were also made with respect to the types of descriptors important for partitioning across the various media. PMID- 12570044 TI - The inorganic and organic characters for predicting bioconcentration on wide variety of chemicals in fish. AB - The applicability of Fujita's inorganic (i) and organic (o) characters as descriptors for predicting bioconcentration factor (BCF) in fish was investigated with a wide variety of organic chemicals. Among 612 BCF data recently released by National Institute of Technology and Evaluation (NITE), the values for the chemicals with a molecular weight of less than 600 and the 1-octanol/water partition coefficient (log P) of less than six were extracted and analyzed. By applying theoretically derived model equation, a good relationship between these BCF and each Fujita's sigma i: sigma o was established. Statistical analyses and model validations revealed that the estimations of our approach were very excellent. More precise predictions were attained than those by using other published models, especially, for chemicals such as disperse dye having hetero atoms. PMID- 12570045 TI - Docking study of cistanoside C to telomeric DNA fragment. AB - Experiments show that the natural products phenyl propanoid glycosides (PPGs) extracted from the plant Pedicularis spicata are capable of repairing DNA damaged by oxygen radicals. Based on kinetic measurements and experiments on tumor cells, a theoretical study of the interaction between PPG molecule Cistanoside C and telomeric DNA fragment has been carried out. The docking calculations performed using JUMNA software showed that the Cistanoside C could be docked into the minor groove of telomeric DNA and form complexes with the geometry suitable for an electron transfer between guanine radical and the ligand. Such complexes can be formed without major distortions of DNA structure and are further stabilized by the interaction with the saccharide side-groups. PMID- 12570046 TI - Novel matrix invariants for characterization of changes of proteomics maps. AB - Previous studies on mathematical characterization of proteomics maps by sets of map invariants were based on the construction of a set of distance-related matrices obtained by matrix multiplication of a single matrix by itself. Here we consider an alternative characterization of proteomics maps based on a set of matrices characterizing local features of an embedded zigzag curve over the map. It is shown that novel invariants can well characterize proteomics maps. Advantages of the novel approach are discussed. PMID- 12570047 TI - PLS-QSAR of the adult and developmental toxicity of chemicals to Hydra attenuata. AB - Autocorrelation descriptors encoding lipophilicity, molar refractivity, the H bonding acceptor and donor ability of the molecules and also indicator variables were used to describe 30 organic chemicals characterized by their adult and developmental toxicities to Hydra attenuata. A PLS regression analysis was successfully employed to derive a QSAR model allowing the simulation of both endpoints. Comparisons were made with orthogonal regression analysis and different nonlinear regression analyses. PMID- 12570048 TI - QSARs for identifying and prioritizing substances with persistence and bioconcentration potential. AB - From the 8511 chemicals with 1998 production volumes reported to the U.S. Environmental Protection Agency (U.S. EPA), the TSCA Interagency Testing Committee's (ITC's) Degradation Effects Bioconcentration Information Testing Strategies (DEBITS) was used to identify 56 chemicals. The DEBITS Quantitative Structure-Activity Relationships (QSARs) and the U.S. EPA's PBT profiler QSARs were used to predict the persistence and bioconcentration factors of these 56 chemicals. Partial order ranking was used to prioritise the chemicals based on persistence and bioconcentration potential. PMID- 12570049 TI - Quantitative molecular similarity analysis (QMSA) methods for property estimation: a comparison of property-based, arbitrary, and tailored similarity spaces. AB - Three classes of arbitrary quantitative molecular similarity analysis (QMSA) methods have been computed using atom pairs, topological indices, and physicochemical properties. Tailored QMSA models have been developed using a selected number of TIs chosen by ridge regression. The methods have been applied to the K-nearest neighbor based estimation of log P of two sets of chemicals. Results show that the property-based and tailored QMSA methods are superior to the arbitrary similarity methods in estimating log P of both sets of chemicals PMID- 12570050 TI - Ranking of volatile organic compounds for tropospheric degradability by oxidants: a QSPR approach. AB - The limited availability and variability of data related to atmospheric degradation reaction is a very relevant issue in studies related to environmental fate and behavior of chemicals. For screening purposes, the experimental data of the oxidation rate constants for the reactions with the radicals OH, NO3 and with ozone of 65 heterogeneous organic compounds were explored by Principal Component Analysis: a ranking of volatile organic chemicals (VOC) according to their relative overall atmospheric degradability and an atmospheric persistence index (ATPIN) is proposed. This index has been modeled by theoretical molecular descriptors to obtain MLR models with high predictive power, both internally and externally validated, and the definition of chemical domain applicability. This procedure allows a fast ranking of VOCs according to their tendency to be degraded by atmospheric oxidants, starting only from the knowledge of their molecular structure. PMID- 12570051 TI - The epidemiology of oral and pharynx cancer in Brazil. AB - A study was carried out on the occurrence of oral and pharynx cancer among populations living in five Brazilian geographic regions, by using incidence and mortality rates and trends. Information on survival was also analyzed. Complementary information regarding prevalence of the disease risk factors has been supplied in the attempt to establish some etiological connections with the incidence and mortality observed. Two clear patterns for the occurrence of oral and pharyngeal cancer have been detected: a pattern for the north (including the northern, northeastern and midwestern regions, which are the poorest ones) and a pattern for the south (including the southern and southeastern regions). The southern pattern revealed incidence rates higher than 15 per 100,000 among males, and the northern pattern displayed lower rate levels. Similarly, mortality rates in the south were higher than three per 100,000 and the northern pattern was characterized by lower rates. Other cancers linked to tobacco smoking, such as esophagus, larynx, lung and bladder cancer kept the same pattern of incidence and mortality rates related to oral and pharyngeal cancer in males per region. Information on survival of patients with oral and pharyngeal cancer was very restricted in Brazil, thus hindering the survival analysis per region, but the studies conducted allowed to conclude that the majority of diagnoses were conducted in advanced stages of the disease and implied the low 5-year survival rate observed in Brazil. Tongue tumors displayed the worst survival rates. The southern region shares specific risk factors involved in the oral and pharyngeal cancer present in the Argentinean and Uruguayan populations, namely heavy tobacco smoking and alcohol consumption, as well as high intake of charcoal-grilled red meat and mate. In addition to these risks, part of the oral and pharyngeal cancer registered in Brazil might be linked to a few factors present among people with low life standards, including poor oral hygiene and nutrition. PMID- 12570052 TI - Prognostic significance of microscopic and macroscopic extracapsular spread from metastatic tumor in the cervical lymph nodes. AB - It has been established that the presence or absence of cervical node metastases in patients with head and neck squamous cell carcinoma (HNSCC) is a powerful prognostic indicator. This report reviews the evolution of thinking over the past 70 years with regard to the import and detection of cervical nodal metastases which exhibit spread of tumor beyond the confines of the original encompassing nodal capsule. In the process, this discussion touches upon clinical examination, gross and microscopic pathologic examination, and radiographic imaging studies. In particular, the distinction between gross nodal extracapsular spread of tumor and microscopic nodal extracapsular spread of tumor has been drawn in recent reports; this raises the possibility that identification of microscopic breaching of the nodc capsule by tumor might provide clinically significant information which is not provided by the gross observation of an intact lymph node capsule. While it remains to be seen whether microscopic extracapsular spread alone will prove to be an important prognostic factor, it is recommended that selective neck dissection continue to be offered even in those patients with clinically negative necks; further studies should aid in defining the import of microscopic extracapsular tumor spread in patients with positive cervical nodes. PMID- 12570053 TI - BcI-xL antisense treatment sensitizes Bcl-xL-overexpressing squamous cell carcinoma cells to carboplatin. AB - Carboplatin (CBDCA) has been widely used for the treatment of oral squamous cell carcinoma (SCC). The Bcl-2 family member Bcl-xL has been demonstrated to provide resistance to chemotherapeutic agents including CBDCA. Morpholino Bcl-xL antisense oligonucleotides (oligos) were employed to down-regulate Bcl-xL in CBDCA-resistant (MIT8, MIT16) as well as CBDCA-sensitive (MIT7) SCC cell lines. The oligos were delivered to adherent cells using a scrape-load procedure. The Bcl-xL antisense reduced Bcl-xL levels without altering the level of control actin, suggesting the specificity of this agent. The addition of Bcl-xL antisense oligos substantially prevented the cell growth of both CBDCA-sensitive and resistant cells. The CBDCA-induced partial prevention of cell growth was further augmented by the addition of the Bcl-xL, but not the control, antisense oligos. The morpholino type Bcl-xL antisense oligos may be useful for the treatment of SCC, especially multidrug-resistant tumors with enhanced Bcl-xL levels. PMID- 12570054 TI - Angiosarcoma of the head and neck with intra-oral presentation. A clinico pathological study of four cases. AB - Angiosarcoma of the head and neck is a rare malignant tumour that most commonly involves the skin and subcutis, particularly of the scalp. It has been reported in intra-oral locations very rarely. This study reports on the clinico pathological features of four such cases, in elderly patients and manifesting an aggressive course. All patients were admitted for rapidly enlarging masses involving the hard palate, alveolar crest and retromolar trigone and displaying bone destruction in three cases. Morphologically, three cases were composed by bundles of spindle cells admixed with variably sized abortive vascular channels which only showed slit-like spaces separating large clusters of epithelioid cells. In all instances, intra-cytoplasmic vacuoles containing intact or fragmented red blood cells were distinctly evident. All neoplasms showed consistent CD 31 immunoreactivity and variable degrees of CD 34 and Factor VIII related antigen positivity and were responsible for the death of the patients, for local progression or lung metastases, within 6 months from the original diagnosis. Due to their rarity, angiosarcomas with intra-oral presentation may be diagnosed with difficulty, especially on small incisional biopsies. and their differentiation from spindle cell carcinoma, malignant melanoma, anaplastic lymphoma, malignant myoepithelioma and other sarcomas that more commonly arise at this site should be based on accurate immunohistochemical characterisation. PMID- 12570055 TI - The presence of candidate tumor suppressor gene loci at chromosome 3p for oral squamous cell carcinomas. AB - We investigated the short arm of chromosome 3 (3p) for allelic imbalances, including loss of heterozygosity (LOH) and microsatellite instability (MSI) in 40 primary oral squamous cell carcinomas (SCCs) using 10 microsatellite markers and constructed a deletion map for this chromosome arm. We examined 40 primary tumor tissues, 40 corresponding normal tissues, and seven lymph node metastatic tissues. LOH at one or more loci was found in 24/40 (60%) of tumors. Deletion mapping of these tumors revealed at least three discrete, commonly deleted regions on the chromosome arm. Furthermore, we detected MSI in six of those tested cases (15%). We compared our results with the clinicopathologic features. A number of sites displaying LOH at 3p could be detected in early stage lesions, and the frequencies of LOH tended to be higher in later clinical stages. Thus, the frequent LOH was observed from early stage in pTNM classification. An unknown tumor suppressor gene in the genesis of oral squamous cell carcinoma may exist in 3p. PMID- 12570056 TI - Expression of beta-catenin in rat oral epithelial dysplasia induced by 4 nitroquinoline 1-oxide. AB - The aim of this study was to evaluate whether beta-catenin accumulation is useful for diagnosing the malignant potential of oral precancerous lesions. We investigated oral epithelial dysplasia adjacent to early cancer induced by 4 nitroquinoline 1-oxide in rats. Localization of beta-catenin and cell proliferation were detected immunohistochemically, and exon 3 of the beta-catenin gene was analyzed. Accumulation of beta-catenin in the cytoplasm and nucleus was evident in 10 of 16 dysplasia lesions. Since almost all of the dysplastic lesions in these rats transformed to invasive cancer, beta-catenin accumulation may contribute to the early stage of carcinogenesis. The Ki-67 labeling index was significantly higher in dysplasia and early cancer than in no change. However, there were no significant differences between the expression patterns of beta catenin protein, suggesting that other proliferation pathways are involved in the early stage of tumor development in addition to beta-catenin accumulation. No mutations of exon 3 of the beta-catenin gene were detected in any of the dysplasia or early cancer lesions. These findings suggested that beta-catenin accumulation in the cytoplasm and nucleus without mutation of exon 3 is an early event during carcinogenesis in this tongue cancer model. PMID- 12570057 TI - Suppression of Egr-1 expression in human oral squamous carcinoma cells by okadaic acid. AB - We examined the expression of early growth response-1 (Egr-1) gene in human oral squamous carcinoma cell lines SCCKN and SCC-25 cells and human osteoblastic cell lines Saos-2 and MG63 cells treated with okadaic acid, a potent inhibitor of protein phosphatases type 1 and type 2A. Western blot analysis revealed that Egr 1 was strongly expressed in SCCKN cells and that okadaic acid decreased the expression of Egr-1 protein in these cells. However, Egr-1 was expressed at lower levels in SCC-25, Saos-2, and MG63 cells and transiently increased with the okadaic acid treatment. Suppression of Egr-1 protein expression in okadaic acid treated SCCKN cells stemmed from the suppression of the Egr-1 mRNA level, as determined by the RT-RCR method. Formaldehyde-fixed and alcohol-permeabilized cultured SCCKN cells were reacted with the anti-Egr-1 antibody using immunohistochemical methods. Intense fluorescence was observed in the nuclei of the control SCCKN cells interacted with anti-Egr-1 antibody. However, only a weak reaction was observed in the nuclei in SCCKN cells treated with okadaic acid. A gel mobility shift assay showed that treatment of SCCKN cells with okadaic acid suppressed Egr-1 binding to the DIG-labeled Egr-1 consensus oligonucleotide probe. The present results indicate that the alteration of phosphorylation states in SCCKN cells regulates Egr-1 binding to its consensus sequence and its expression at the transcriptional level. PMID- 12570058 TI - Osteosarcoma arising in Paget's disease of the mandible. AB - Although osteosarcoma is a well-known complication of Paget's disease of bone, it uncommonly develops in the jaw bones. We present an osteosarcoma arising in Paget's disease of the mandible with unique features of a normal serum alkaline phosphatase level, and histologic features of telangiectatic change in the osteosarcoma and association with cemento-osseous dysplasia. Sixteen reported cases of osteosarcoma arising in Paget's disease of the jaw bones (OPJ) are also reviewed and compared to osteosarcoma arising in Paget's disease occurring in the entire skeleton (OPS) and osteosarcoma arising de novo in the jaw bones (OJ). Females are more commonly involved in OPJ in contrast to a male predominance in OPS and OJ. OPJ also has a distinctively higher percentage involving blacks compared to OPS. The prognosis of OPJ is poor, with 69% of patients dying within two years after diagnosis. Early recognition, early and aggressive treatment are important to improve the prognosis and are hence emphasized. PMID- 12570059 TI - Intra-osseous soft tissue perineurioma of the inferior alveolar nerve. AB - This report describes an intra-osseous perineurioma affecting the inferior alveolar nerve, the second at this site, of a 53-year-old man. It produced a well defined, partially corticated, round radiolucency associated with resorption of the root apices of 36 and 37. Histology showed a circumscribed, partially encapsulated cellular tumour composed of bland spindle cells with undulating nuclei and indistinct cell membranes arranged in interlacing fascicles. The tumour was strongly positive for epithelial membrane antigen, collagen IV and laminin, with only scattered S100-positive cells and lack of CD34 expression. Electron microscopy showed elongated cells with continuous external laminae of basement membrane-like material, subplasmalemmal densities, desmosome-like junctions, and variable numbers of micropinocytotic vesicles. The lesion was surgically excised and in 9 months there has been no recurrence. PMID- 12570060 TI - A needle free system for cervical lymphatic mapping and sentinel node biopsy in oral squamous cell carcinoma. AB - The techniques of lymphatic mapping and sentinel node biopsy are finding increasing utility in the management of the loco-regional lymphatics in many areas of oncology. Our unit is currently investigating their feasibility in relation to the management of the clinically node negative neck in oral squamous cell carcinoma. In order to reduce patient discomfort, and to remove the possibility of sharps injury, particularly in the high-risk patient, we report the use of a needle free device for the intra-oral delivery of the radiopharmaceutical. We believe that this is the first report using such a device in the oral cavity for the purpose of lymphoscintigraphy, and that it has significant advantages over the conventional hypodermic needle in certain patient groups. PMID- 12570061 TI - Odontoameloblastoma. Clinico-pathologic study of three cases and critical review of the literature. AB - The odontoameloblastoma (OA), is an infrequent neoplasm. To date, there are less than 50 cases reported as OA or ameloblastic odontoma in the English dental literature, but only 14 (including three of our own material), fulfill the histological criteria of the current WHO histological classification of odontogenic tumours. Nine occurred in men and five in women (male to female ratio 1.8:1). Age ranged from 2 to 50 years (mean 20.2 years), and nine cases (64.2%) were diagnosed during the first two decades. Maxilla and mandible were equally involved, and most cases occurred posterior to the canines (71.4%). Follow-up ranged from 6 months to 8 years (mean: 25.5 months). Of the 12 cases with informed follow-up, two recurred once (at 24 and 18 months, respectively), and one case had two documented recurrences, at 6 and 49 months. Although OA tends to occur at an earlier age than conventional ameloblastoma, it has practically the same potential to produce bone expansion, root resorption and recurrence. For these reasons OA should be treated in a similar fashion, with wide surgical excision and close follow-up for at least 5 years. PMID- 12570062 TI - Comment on: F. Sandra et al. "The role of MDM2 in the proliferative activity of ameloblastoma", oral oncology 2002;38(2): 153-57. PMID- 12570063 TI - Odontomas and ameloblastomas: variable prevalences around the world? PMID- 12570064 TI - Comment on: Kuffer and Lombardi "Premalignant lesions of the oral mucosa. A discussion about the place of intraepithelial neoplasia", Oral Oncology 2002;38:125-30. PMID- 12570065 TI - When is an oral leukoplakia premalignant? PMID- 12570066 TI - Atypical antipsychotics in mood disorders. AB - Bipolar disorder is ranked as the sixth most important worldwide cause of disability. Current treatment is based chiefly on lithium and/or anticonvulsants, of which sodium valproate is the most widely used. A significant minority of patients fail to respond fully to current treatments, particularly those with mixed mania and/or rapid cycling. Many patients are unable to tolerate the side effects of current therapy in the long term, and adverse effects may contribute to the high rate of noncompliance observed in bipolar disorder. The shortcomings of current treatments are reflected in poor outcomes: two-thirds of patients with bipolar disorder require hospitalization on more than one occasion; employment and social functioning are significantly lower than in control groups; 93% of carers suffer at least moderate distress; and 25-50% of patients are believed to attempt suicide at least once. Bipolar disorder shares some features with schizophrenia, and several atypical antipsychotics have demonstrated efficacy in bipolar disorder. Quetiapine has a particularly favourable tolerability profile, with placebo-level extrapyramidal symptoms and prolactin levels across the entire dose range combined with a neutral effect on weight during long-term use, and may be a valuable treatment option in acute mania and bipolar disorder. PMID- 12570067 TI - Schizophrenia and bipolar disorder: similarities in pathogenic mechanisms but differences in neurodevelopment. AB - Over the past 100 years, the Kraepelinian classification of psychoses has dominated our approach to schizophrenia and bipolar disorder. However, controversy as to the nature of the illnesses--whether they can be viewed as completely distinct, essentially the same, or occupying different points along a psychosis spectrum--has intensified in recent years. This paper reviews the evidence for these differing opinions, examining both the commonalities between the two diseases and the distinctions. A genetic propensity towards psychotic disorders is widely acknowledged; more recent studies suggest a considerable overlap in genetic susceptibility to schizophrenia and bipolar disorder. The influence of early environmental effects, such as obstetric complications, on schizophrenia is also established but little such evidence exists for bipolar disorder. Structural abnormalities of the brain of developmental origin as well as neuropsychological deficits have been clearly identified in schizophrenia but less evidence has been found in bipolar disorder. The most plausible explanation is that one or more susceptible genes are shared between schizophrenia and bipolar illness, and can be thought of as predisposing individuals to psychosis, perhaps by producing a dysregulation of the dopaminergic response to stress. Other genes and environmental factors are likely to have more specific effects and contribute to producing the patterns that psychiatrists recognize as 'classical' schizophrenia and mania. In particular, genes involved in early cortical development and early neurodevelopmental insults causing developmental impairment may put individuals on a trajectory towards schizophrenia rather than bipolar illness. PMID- 12570068 TI - Mood stabilization and the role of antipsychotics. AB - The pharmacological management of bipolar disorder is complex as a result of the cyclic nature of the condition. Long-term treatment is, however, essential to control the symptoms of depression and mania and to stabilize the cyclical mood changes. In particular the use of mood stabilizers in all phases of treatment has been acknowledged. The atypical antipsychotics are being increasingly used to control acute manic episodes, and data are emerging to support their mood stabilizing and antidepressant properties. This article will review the results from open and double-blind studies with quetiapine, ziprasidone, risperidone and olanzapine in the management of bipolar disorder. PMID- 12570069 TI - Choosing an atypical antipsychotic. AB - The atypical antipsychotics vary in terms of their pharmacological profiles, particularly in relation to their tolerability, effects on safety parameters and patient acceptability. Olanzapine, risperidone and ziprasidone are associated with extrapyramidal symptoms (EPS) in a dose-dependent manner, whereas quetiapine has been shown to produce a significantly lower incidence of substantial EPS effects than haloperidol, and less EPS requiring treatment than risperidone. Similarly, unlike risperidone and haloperidol, quetiapine treatment has been associated with a significant reduction in serum prolactin levels, and has normalized raised prolactin levels after discontinuation of previous treatment. Weight gain is also one of the major unwanted adverse effects of treatment with many antipsychotic drugs but, in contrast, quetiapine has demonstrated a neutral or 'normalizing' effect on body weight. In comparison with other antipsychotics, quetiapine has been shown to possess a favourable safety profile, with no requirement for routine blood, thyroid, or liver monitoring during treatment. Overall, quetiapine therapy has produced high levels of patient satisfaction and compliance and this, coupled with its efficacy in reducing psychoses of various origins, has made it an attractive treatment option in both patients at increased risk of EPS, and the general population. PMID- 12570070 TI - Furniture strippers need to adopt a pollution prevention approach. PMID- 12570072 TI - AIHA expanded position statement on extremely low frequency electric and magnetic fields. PMID- 12570071 TI - More on Trinity. PMID- 12570073 TI - AIHA white paper on extremely low frequency (ELF) fields. PMID- 12570074 TI - Partition coefficients for benzene in human skin. AB - The contribution of benzene to body burden after skin absorption compared with that due to inhalation absorption is of potential interest in the setting and interpretation of benzene (inhalation) exposure standards. However, an understanding of the quantitative relationship between skin and inhalation absorption, under different exposure conditions, is required. Such knowledge may be gained through physiological based pharmacokinetic (PBPK) modeling. The intake of benzene to the body via inhalation has been studied extensively. Physiological parameters enabling the calculation of amounts of benzene entering the blood stream per unit time are readily available for use in a PBPK model. Unfortunately, some data (i.e., partition coefficients) that would enable biologically plausible calculation of amounts of benzene entering the blood stream via skin absorption in a PBPK model are not available. Hence, the aim of this research was to determine partition coefficients across the epidermal and dermal layers of human skin so that these could be used within a PBPK model to determine quantitatively the flow rate of benzene per unit time through intact skin into the blood stream. The partition coefficients found for blood substitute: viable epidermis and blood substitute: dermis were, respectively, 2.4 and 11.2. Partition coefficients for benzene : stratum corneum (4.2), whole skin : blood substitute (2.2), benzene : water (109/126), and benzene : blood substitute (55/59) also were determined for the purposes of validating the blood substitute: viable epidermis and blood substitute : dermis partition coefficients. PMID- 12570075 TI - Dynamic monitoring of the dust pickup efficiency of vacuum cleaners. AB - This study evaluated a new method that uses an optical aerosol photometer for dynamically monitoring dust pickup efficiency during vacuuming. In the first stage of this study the new method was compared with built-in dirt sensors installed by vacuum cleaner manufacturers. Through parallel testing it has been shown that the widely available built-in dirt sensors are not sensitive enough to register small (< 53 microm) dust particles. Therefore, only the optical photometer was used in the rest of the experiments of this study to monitor the dust pickup efficiency while the vacuum cleaner was operated with different nozzles on clean and soiled carpet and vinyl sheet flooring. This method also was used to monitor dust pickup efficiency when vacuuming carpets originating from lead-contaminated homes. The dust pickup efficiencies obtained with the optical aerosol photometer have been compared with the surface lead concentrations found during different stages of cleaning. Results indicate that the dust mass concentration registered with the optical aerosol photometer at the nozzle outlet correlates well with the dust mass collected in the vacuum cleaner filter bag and with the surface lead level. Therefore, dynamic dust pickup monitoring can provide valuable information about the efficiency of cleaning when a vacuum cleaner is used. This suggests that a small aerosol photometer similar to a light scattering smoke detector would be beneficial in vacuum cleaners used for cleaning surfaces contaminated with leaded dust and biological particles (including allergens). PMID- 12570076 TI - Filter and cassette mass instability in ascertaining the limit of detection of inhalable airborne particulates. AB - In the gravimetric assessment of workplace aerosols, there is an increasing need to confidently measure smaller and smaller collected masses. To do this, it is important to know both the limit of detection (LOD) and limit of quantification (LOQ) of the analysis performed, determined by the weighing imprecision of blank samples. Of particular current interest is the measurement of inhalable aerosols, as defined for many substances in the American Conference of Governmental Industrial Hygienists threshold limit values list. One popular method is the use of a filter contained within a small cassette, in which both are weighed. Earlier investigations of plastic and stainless steel cassettes showed that plastic cassettes are highly sensitive to changes in humidity. But one study also demonstrated that the resulting changes in mass could be largely corrected using field blanks. An investigation, therefore, was undertaken to determine the weighing imprecision of various cassette and filter assemblies, assuming blank corrections are made. Three types of filter (Teflon, glass fiber, and polyvinyl chloride) were investigated in combination with three types of cassette (plastic, nickel-plated plastic, and stainless steel). Results show that regardless of the substrate being used, sample masses equal to or higher than 0.19 and 0.65 mg can be confidently detected and quantified, respectively. PMID- 12570077 TI - VOC exposures in a mixed-use university art building. AB - Despite a sizable educational art enterprise in the United States there is a dearth of rigorously performed studies of exposures to persons engaged in such activities. Exposures to 45 EPA-designated volatile organic compounds were examined in printmakers in a mixed-use university art school served by a 100% exhausted mechanical ventilation system. Personal exposures (n = 90) were compared with area concentrations (n = 36) in the studio area and at a second location at the same facility. For personal exposure assessments a cohort of 12 students wore passive dosimeters twice weekly over a 6-week period. Numerous compounds were found, the most prevalent being toluene at an average concentration of 64.6 microg/m3 (17.1 ppb; range < 1-319 ppb); 1,1,1, trichloroethane at 40.5 microg/m3 (7.5 ppb; range < 1-211 ppb); xylenes at 8.0 microg/m3 (1.8 ppb; range < 1-43 ppb); 1,3,5-trimethyl benzene at 6.2 microg/m3 (1.3 ppb; range < .3-32 ppb); propyl benzene at 5.0 microg/m3 (1.0 ppb; range < .5-27 ppb); methylene chloride at 4.9 microg/m3 (1.4 ppb; range < 1-10 ppb); and ethyl benzene at 4.5 microg/m3 (1.1 ppb; range < .4-23 ppb). Personal exposures were considerably higher than average area air concentrations, with the exception of methylene chloride concentrations, which were five times higher at the print cleaning operation. Floors where solvents were not used had no detectable exposures (typical lower limit of detection approximately 1 ppb) and were free of solvent odors. Despite frequent solvent contact with skin, personal protective equipment was seldom used. Results indicate that in mixed-use facilities such as this, nonrecirculating general ventilation systems can effectively eliminate indoor air quality issues between floors, despite perceptible odors on solvent use floors. For total exposure assessments in such processes, contact exposures from printmaking solvents during cleaning procedures are a potentially important consideration. PMID- 12570078 TI - Exposure levels and determinants of softwood dust exposures in BC lumber mills, 1981-1997. AB - Measurements of personal exposure to wood dust (n = 1237) collected by the Workers' Compensation Board of British Columbia, Canada, over the period 1981 1997 were used to construct an empirical model to identify broad determinants of softwood dust exposure. Potential determinants of exposure examined included species of tree processed; company; geographic location of lumber mill; department; job title; calendar year; and production factors such as board feet of lumber produced per year. A determinants of exposure model was built using multiple linear regression. Nested within this compliance database was a subset of samples collected for a research study. These enabled the authors to explore whether differences in exposure measurements can in part be explained by sampling strategy (research versus compliance). Potential differences were examined by examining differences in means for each job title, stratified by sampling strategy; and by offering "sampling strategy" as a categorical predictor variable to the empirical model. Multiple linear regressions revealed the most important determinants of increased wood dust exposure to be mill location away from the coast, earlier calendar year, and indoor jobs. The empirical model had an R2 of 0.39 and a predictive range from 0.02 to 25.45 mg/m3. Research and compliance sampling strategies showed no difference in mean exposure and distribution in the empirical model (p < 0.05), suggesting that regulatory exposure databases may be of utility for exposure assessment in epidemiology. This research indicates that compliance-sampling strategies do not result in an overestimation of mean exposure levels within jobs, but they do focus on a biased sample of jobs-those most highly exposed. PMID- 12570079 TI - Development of a sampling and analytical method for 2,2-dichloro-1,1,1 trifluoroethane in workplace air. AB - The use of 2,2-dichloro-1,1,1-trifluoroethane (compound number: HCFC-123) is growing in industry as a substitute for ozone-depleting chlorofluorocarbons (CFCs). Recently, liver-related illnesses have been reported from industries handling HCFC-123. However, information on worker exposure to the material is limited, and an acceptable sampling/analytical method is not available. The aim of this study was to develop a widely applicable sampling and analytical method to determine worker exposures to airborne HCFC-123 and to evaluate the performance of the method. A solid sorbent tube, containing two sections (400 mg in the front and 200 mg in the back) of activated coconut-shell charcoal was chosen for sampling airborne HCFC-123 vapor. The breakthrough volumes were 13.6 L at 3597 +/- 210.1 ppm (with a sampling airflow rate of 0.046 L/min) and 17.0 L at 1841 +/- 4.5 ppm (with sampling airflow rate of 0.046-0.050 L/min). Samples of HCFC-123 in the charcoal tube were stable for 7 days either at room temperature or in a refrigerator and a migration occurred within 14 days at room temperature. It is recommended that the HCFC-123 sample in activated charcoal tubes be stored either at room temperature or in a refrigerator and be analyzed within 7 days. The HCFC-123 in the charcoal tubes was desorbed into dichloromethane and analyzed using gas chromatography/ flame ionization detection. The limit of detection was 0.23 mg/sample, and the average desorption efficiency was 99.0%. The total coefficient of variation was 0.060, and the method accuracy was 16.6%. In conclusion, the performance of the sampling and analytical method developed for the determination of airborne HCFC-123 concentrations was acceptable to the NIOSH sampling and analytical criteria. PMID- 12570080 TI - Investigation of the presence of amino and nitro polycyclic aromatic hydrocarbons in a Soderberg primary aluminum smelter. AB - An excessive risk of bladder cancer among Soderberg primary aluminum smelter potroom workers has been reported by several authors, and to date, a causal agent has not been identified. Certain nitro and amino polycyclic aromatic hydrocarbons (PAHs) are either known or suspected human bladder carcinogens, and their presence in this work environment would be cause for concern. In fact, one of these, 2-aminonaphthalene, has been found in both the air and the coal tar pitch used to fabricate process electrodes following an investigation in a Soderberg plant. During the present study, monitoring was conducted in a primary Soderberg aluminum smelter over a 3-week period using novel methods having very good specificity, reproducibility, and sensitivity. Results indicated that 1 aminonaphthalene and 2-aminonaphthalene and 1-nitronaphthalene and 2 nitronaphthalene were present at low parts-per-million levels in the coal tar pitch used to prepare the process electrodes. 1-aminonaphthalene and 2 aminonaphthalene were measured 40 cm above the electrolytic cells on days 1 and 2 of the investigation when the cells' ventilation systems had been shut down for repairs and maintenance. 2-aminonaphthalene was measured at concentrations ranging from 0.080 to 0.350 microg/m3. 1-nitronaphthalene and 2-nitronaphthalene, predominant in coal tar pitch, were not detected in any of the 43 air samples collected. A plausible explanation is their reduction to their respective amine during the electrolytic process. The routine screening of coal tar pitch used to prepare the process's electrode for both amino and nitro PAHs should be adopted by the aluminum industry as an effective means of controlling the presence of these carcinogenic pollutants in their workplace. PMID- 12570081 TI - Monitoring for N,N-dimethylformamide and N,N-dimethylacetamide with a diffusive sampler using distilled water as an absorbent. AB - To obtain an appropriate sampling method for evaluating individual workers' exposure concentrations to chemicals highly soluble in water, the suitability of LiPS (liquid passive sampler) as diffusive sampler was examined. Although adsorbed chemical substances with activated carbon felt being used as the adsorbent must be desorbed in such samplers, this process can be omitted by using liquid as the absorbent. This kind of simplification can be utilized for water soluble solvents such as N,N-dimethylformamide (DMF) and N,N-dimethylacetamide (DMAC). In laboratory experiments DMF and DMAC levels determined by this method were sufficiently correlated with exposure concentrations, exposure duration, and cumulative exposure levels. The sampling rates for DMF and DMAC obtained from these results were 0.50 and 0.43 mL/min, respectively, and the theoretical sampling rate for DMF obtained using Fick's diffusion formula was consistent with the experimental value. The effects of relative humidity, wind velocity, and temperature on the measured values were negligible within the range expected in a practical industrial environment. In a field survey the exposure concentrations for 38 workers handling DMF in the process of manufacturing spandex fiber and polyurethane gloves were determined both by this method and by the National Institute of Occupational Safety and Health (NIOSH) method, in which silica gel tubes are used. The DMF concentrations detected by the two methods were comparable (Y = 0.912X, correlation coefficient: 0.932). This LiPS method proved to be useful in determining the concentrations of DMF and DMAC to which workers were exposed. PMID- 12570082 TI - Airborne fiber exposure assessment of dry asbestos-containing gaskets and packings found in intact industrial and maritime fittings. AB - This study assessed airborne fiber exposures from intact asbestos-containing gaskets and packings while activities representative of historical work practices were performed. The study design eliminated analytical interferences while systematically capturing information related to activity variables. A series of gasket and packing activities was conducted according to traditional methods while sampling was performed to determine the 8-hour time-weighted average (TWA). The fittings used during this study were obtained intact from a decommissioned industrial power plant and U.S. Navy destroyers. The activities tested included flat blade scraping, hand wire brushing, power wire brushing, making gaskets with a ball-peen hammer, and stem packing removal and replacement. All activities were performed dry. Results for every area and personal sample showed the 8-hour TWAs were well below the current Occupational Safety and Health Administration permissible exposure limit of 0.1 f/cc. A database of more than 400 points was developed to analyze information from variable factors related to the tests. These factors included, for example, type of gasket, composition of the gasket, percentage of gasket adhering to a flange surface, gasket surface area, and minutes elapsed for removal. The results demonstrate a very low rate of fiber exposure from routine activities associated with asbestos-containing elastomeric gaskets and impregnated packing. PMID- 12570083 TI - Chemical characterization of sanding dust and methylene chloride usage in automotive refinishing: implications for occupational and environmental health. AB - Surface preparation activities conducted during automotive refinishing present several potential human health and environmental risks. This study examines the chemical composition of vehicle sanding dust and the prevalence of methylene chloride use as a basis for evaluating potential chemical exposures in the work environment, fugitive environmental releases, and take-home toxics. This article reports on the findings of (1) a statewide technology and work practices survey of 353 licensed auto body shops and (2) laboratory analyses of sanding dust representing more than 200 vehicles, 10 commercial body filler compounds, and work shirts worn during vehicle sanding while using nonventilated equipment. Survey data revealed that the majority of shops (78%) do not use ventilated sanding equipment, that most workers (55%) take their work clothes and shoes home at the end of the workday, and that 17% of the respondents used a methylene chloride-based paint stripper as an adjunct to mechanical sanding. Laboratory results showed that Pb, As, Cr, Mn, and Ni were present in the sanding dust at every facility tested. Lead concentrations in sanding dust were found to be highest at facilities that performed complete vehicle refinishing (range 770 to 7300 ppm) and at a collision repair shop that used a high-lead content body filler compound (1800 ppm). Hexavalent chromium also was found in two vocational high school paint dust samples at concentrations of 54 and 710 ppm. When total lead and chromium concentrations reached 7300 and 2300 ppm, respectively, facility sanding dust samples failed the U.S. Environmental Protection Agency's Toxicity Characteristic Leaching Procedure for hazardous waste. Metals found in the sanding dust also were present on the work shirts of technicians-ranging from 0.06 (Cd) to 81 (Mg) microg/inch2 of cloth-who sanded on paint without ventilated equipment. Results suggest that sanding dust and methylene chloride paint strippers used in vehicle resurfacing operations pose a potential hazard to human health and the environment. PMID- 12570084 TI - Exposure to respirable crystalline silica in eastern North Carolina farm workers. AB - Occupational exposure to crystalline silica has been linked to silicosis, some forms of cancer, and certain autoimmune diseases. Little information exists on exposure levels of respirable silica in the agricultural industry. This study assessed respirable silica exposure of farm workers in eastern North Carolina. Sandy soils in this region have been shown to contain high levels of respirable silica. Personal breathing zone samples (n = 37) were collected from 27 workers at seven farms during various agricultural activities. The highest respirable silica concentrations were measured during sweet potato transplanting (3.91 +/- 2.07 mg/m3). Respirable silica exposure was observed to be associated with agricultural activity, soil moisture, relative humidity, and wind speed. Most of the variation in exposure (79%) was explained by agricultural activity and soil moisture. The observed percentage of silica levels (mean 34.7%) were almost twice as high as was reported in studies of California agriculture. This may be due to the loamy sand and sandy loam soil types in the regions in this study. In agriculture, respirable silica exposure is highly variable, but the potential for exposures above the threshold limit value of 0.05 mg/m3 exists during particular agricultural activities. PMID- 12570085 TI - Comparison of fiber counting by TV screen and eyepieces of phase contrast microscopy. AB - This study designed a modified light path of a phase-contrast microscope to evaluate the feasibility of fiber counting on a television screen (TVS). A comparison also was made of the fiber counts, fiber density, and precision derived from repeatedly counting fibers through eyepieces (EPs) and on a TVS connected to a phase contrast microscope. Thirty asbestos fiber samples were counted 10 times repeatedly. Ten samples were counted by viewing the same field through an EP and on a TVS alternately, whereas the other 20 samples were counted separately by using an EP and a TVS. The A rules of National Institute for Occupational Safety and Health (NIOSH) 7400 method was quoted. No statistically significant difference of fiber density (p = 0.39) or fiber counts (p > 0.05) was observed between the TVS and EP methods, though TVS gave a slightly lower value than EP did. The bias of the two methods was 7.7 +/- 8.0% on an average. The relative standard deviations (RSDs) for both methods were the same, 0.40, which resembled the theoretical RSD, 0.44, of NIOSH 7400. Meanwhile, the theoretical and experimental RSDs were not significantly different for either method (p > 0.05). The modified light path of a phase contrast microscope provided a compatible view with less eye strain on a TVS than a conventional EP. Moreover, operator biases and variability might be greatly reduced by training several counters simultaneously on the TVS. PMID- 12570086 TI - Comparison of three commercially available fit-test methods. AB - American National Standards Institute (ANSI) standard Z88.10, Respirator Fit Testing Methods, includes criteria to evaluate new fit-tests. The standard allows generated aerosol, particle counting, or controlled negative pressure quantitative fit-tests to be used as the reference method to determine acceptability of a new test. This study examined (1) comparability of three Occupational Safety and Health Administration-accepted fit-test methods, all of which were validated using generated aerosol as the reference method; and (2) the effect of the reference method on the apparent performance of a fit-test method under evaluation. Sequential fit-tests were performed using the controlled negative pressure and particle counting quantitative fit-tests and the bitter aerosol qualitative fit-test. Of 75 fit-tests conducted with each method, the controlled negative pressure method identified 24 failures; bitter aerosol identified 22 failures; and the particle counting method identified 15 failures. The sensitivity of each method, that is, agreement with the reference method in identifying unacceptable fits, was calculated using each of the other two methods as the reference. None of the test methods met the ANSI sensitivity criterion of 0.95 or greater when compared with either of the other two methods. These results demonstrate that (1) the apparent performance of any fit-test depends on the reference method used, and (2) the fit-tests evaluated use different criteria to identify inadequately fitting respirators. Although "acceptable fit" cannot be defined in absolute terms at this time, the ability of existing fit-test methods to reject poor fits can be inferred from workplace protection factor studies. PMID- 12570087 TI - Construction noise: exposure, effects, and the potential for remediation; a review and analysis. AB - More than one-half million construction workers are exposed to potentially hazardous levels of noise, yet federal and state Occupational Safety and Health Administration (OSHA) programs provide little incentive to protect them against noise-induced hearing loss. Construction noise regulations lack the specificity of general industry noise regulations. In addition, problems that characterize the construction industry, such as worker mobility and the large proportion of small businesses, make implementing hearing conservation measures more difficult. The apparent severity of exposure depends greatly on the measurement method, with the 3-dB exchange rate almost always showing higher average exposure levels than the 5-dB (OSHA) rule. Construction workers demonstrate hearing threshold levels that generally conform to those expected in manufacturing. The prevalence of hearing protection device (HPD) use among U.S. construction workers is very poor, partly because of perceived difficulties in hearing and understanding speech communication and warning signals. In addition, masking by noise of necessary communication and warning signals is of particular concern in construction, where recent research demonstrated the association between fatalities and the failure to hear reverse alarms. Judicial use of HPDs is of the utmost importance, along with avoiding overattenuation, selecting HPDs with uniform attenuation, and using noise-attenuating communication systems when possible. A successful hearing conservation program in British Columbia can serve as a model for the United States, with a long-standing positive safety culture, a high percentage of HPD use, improvement in average hearing threshold levels over the last decade, and a centralized record-keeping procedure, which helps solve the problem of worker mobility. However, controlling construction noise at the source is the most reliable way to protect worker hearing. U.S. manufacturers and contractors should benefit from the activities of the European Community, where noise control and product labeling in construction has been carried out for more than 20 years. PMID- 12570088 TI - Field comparison of impingers and treated filters for sampling of total aliphatic isocyanates with the MAP reagent. AB - Results of a comparative field study on the performance of 25-mm inhalable samplers (Institute of Occupational Medicine [IOM]) and midget impingers for the collection of total isocyanates in air using the 1-(9 anthracenylmethyl)piperazine (MAP) reagent are presented. Air sampling and analysis was performed according to the National Institute for Occupational Safety and Health MAP draft method 5525. Midget impingers filled with 15 mL of 1 x 10(-4) M MAP in butyl benzoate were operated at 1 L/min. IOM cassettes loaded with 25-mm quartz fiber filters impregnated with 500 microg MAP, were operated at 2 L/min. Filters were field extracted with 10 mL of 1 x 10(-4) M MAP in acetonitrile. Thirty-four impinger-IOM pairs were collected in three autobody shops during spray painting tasks. Regression analysis [Ln (IOM) = 1.0 Ln (impinger), R2 = 0.98] and a paired t-test (Pr > 0.9) demonstrated that impingers and IOMs perform equally in their collection efficiency for the monomer and total oligomeric hexamethylene diisocyanate (HDI). IOM performance did not deteriorate at longer sampling times compared with the impinger performance. Within-sampler variability was calculated from the individual coefficients of variation (CV) of the 17 pairs of like samplers. The mean of CVs (SD)% for the monomer and total oligomeric HDI was approximately 12 (12)% and 15 (13)% for the IOM and the impinger sampler, respectively. Poor correlation (Pearson correlation coefficient < 0.3) and statistically nonsignificant differences (P > 0.74 two-sided) were found for the between-sampler CVs. Factors that might have influenced the observed sampler agreement are also discussed. It is concluded that MAP impregnated filters can be successfully used for sampling of slow curing total aliphatic isocyanates in air. PMID- 12570089 TI - PAT Program report: background and current status. PMID- 12570090 TI - Hypnosis, differential expression of cytokines by T-cell subsets, and the hypothalamo-pituitary-adrenal axis. AB - This investigation tested the hypothesis that hypnosis can differentially modulate T-cell subsets, and that this effect is mediated by changes in hypothalamo-pituitary-adrenal (HPA) mediators. Seven healthy, highly hypnotizable volunteers participated in three one-day sessions, a baseline and two intervention sessions. Hypnosis intervention entailed a standardized induction, suggestions for ego strengthening and optimally balanced functioning of the immune and neuroendocrine systems, and post-hypnotic suggestions for stress management and continued optimal balance of bodily systems. Blood samples were drawn at five time points between 8:00 a.m. and 3:00 p.m. and were analyzed for T cell activation and intracellular cytokine expression (Interferon (IFN)-gamma, Interleukin-2, Interleukin-4) and HPA axis mediators (ACTH, cortisol, and beta endorphin). Following hypnosis intervention, statistically significant immunological effects were noted. Specifically, the proportion of T-cells expressing IFN-gamma (p = .0001) and IL-2 (p = .013) were lower after hypnosis. T cell activation response to polyclonal stimulation was positively correlated with ACTH (p = .01) and beta-endorphin (p = .001) while IFN-gamma expression was correlated with levels of cortisol (p < .001). Further controlled studies utilizing hypnosis with patients in treatment are warranted in order to examine whether an altered T-cell response can be replicated in the presence of disease. PMID- 12570091 TI - Gene expression, neurogenesis, and healing: psychosocial genomics of therapeutic hypnosis. AB - The historical lineage of therapeutic hypnosis in James Braid's "psychophysiology", Pierre Janet's "physiological modification", and Milton Erickson's "neuro-psycho-physiology" is extended to include current neuroscience research on activity-dependent gene expression, neurogenesis, and stem cells in memory, learning, behavior change, and healing. Three conditions that optimize gene expression and neurogenesis--novelty, environmental enrichment, and exercise -could integrate fundamentals of the theory, research, and practice of therapeutic hypnosis. Continuing research on immediate-early, activity-dependent, behavior state-related, and clock gene expression could enhance our understanding of how relaxation, sleep, dreaming, consciousness, arousal, stress and trauma are modulated by therapeutic hypnosis. It is speculated that therapeutic and post hypnotic suggestion could be focused more precisely with the time parameters of gene expression and neurogenesis that range from minutes and hours for synthesizing new synapses to weeks and months for the generation and maturation of new, functioning neurons in the adult brain. PMID- 12570092 TI - Utilization Sobriety: brief, individualized substance abuse treatment employing ideomotor questioning. AB - This article presents a substance abuse treatment method that acknowledges and accommodates the personal needs that are being addressed by substance. This method, Utilization Sobriety, bypasses perceived resistances and employs idiosyncratic psychobiological learning to achieve a body-mind gestalt that is complementary to the client's sobriety. It develops a safe framework for addressing any subsequent mental health themes directly or indirectly related to substance misuse. A treatment protocol for the use of Utilization Sobriety as well as relevant clinical material illustrating its application and a discussion of its implications are offered. PMID- 12570093 TI - Hypnosis-facilitated relaxation using self-guided imagery during dermatologic procedures. AB - Many patients experience some degree of anxiety and/or discomfort during dermatologic procedures. For most patients this anxiety or discomfort is tolerable, but a few find it intolerable to the point of interference with the accomplishment of the procedure. A case is presented in which a 51 -year-old female experienced so much anxiety that it jeopardized the continuation of a necessary procedure. When (with her consent) a trance state was induced through hypnosis, she relaxed and remained in a pleasant mental condition for the remainder of the procedure. This case illustrates the usefulness of hypnosis in selected situations where it can alleviate anxiety and discomfort associated with dermatologic procedures. Suggested trance induction, maintenance, and termination scripts are included to assist those with adequate training and experience in providing this comfort to selected patients through hypnosis. PMID- 12570094 TI - A character in search of character: Narcissistic Personality Disorder and Ego State Therapy. AB - The individual diagnosed with Narcissistic Personality Disorder presents with grandiosity, extreme self-involvement, and lack of interest in and empathy for others. This paper reviews current theories concerning the development and treatment of Narcissistic Personality Disorder, and introduces the use of Ego State Therapy for its treatment. The ego state model of treatment will be described and demonstrated with case material. Initially ego states that reveal the grandiosity will be accessed. As therapy progresses, ego states that hold the underlying feelings of emptiness, rage, and depression are able to emerge. With further treatment, transformation and maturation of the ego states occur, reflecting the changes in internal structure and dynamics as well as improvement in external interpersonal relationships. Issues concerning Ego State Therapy as utilized with personality disorders will be discussed and contrasted with more traditional methods of treatment. PMID- 12570095 TI - Playful metaphors. AB - The inner world of the child is a community of archetypes potentially available for the child's healthy ego development. Many forces limit and prohibit their utility. Play therapy in the context of a hypnotic relationship can potentiate these archetypes into becoming "playful metaphors" for healing and strengthening ego development. In this article, the author describes her use of playful metaphors in her therapeutic work with two children and explores how metaphor in play therapy is able to connect the child with healing archetypal imagery. PMID- 12570096 TI - Magnitude estimation of conceptual data dimensions for use in sonification. AB - Sonifications must match listener expectancies about representing data with sound. Three experiments showed the utility of magnitude estimation for this. In Experiment 1, 67 undergraduates judged the sizes of visual stimuli and the temperature, pressure, velocity, size, or dollars they represented. Similarly, in Experiment 2, 132 listeners judged the pitch or tempo of sounds and the data they represented. In both experiments, polarity and scaling preference depended on the conceptual data dimension. In Experiment 3, 60 listeners matched auditory graphs to data created with the results of Experiment 2, providing initial validation of scaling slopes. Magnitude estimation is proposed as a design tool in the development of data sonifications, with the level of polarity preference agreement predicting mapping effectiveness. PMID- 12570097 TI - Models of performance in learning multisegment movement tasks: consequences for acquisition, retention, and judgments of learning. AB - Participants learned different keystroke patterns, each requiring that a key sequence be struck in a prescribed time. Trials of a given pattern were either blocked or interleaved randomly with trials on the other patterns and before each trial modeled timing information was presented that either matched or mismatched the movement to be executed next. In acquisition, blocked practice and matching models supported better performance than did random practice and mismatching models. In retention, however, random practice and mismatching models were associated with superior learning. Judgments of learning made during practice were more in line with acquisition than with retention performance, providing further evidence that a learner's current ease of access to a motor skill is a poor indicator of learning benefit. PMID- 12570098 TI - Measurement of individual differences in children's suggestibility across situations. AB - The authors attempted to use scores on the Video Suggestibility Scale for Children (VSSC, M. H. Scullin & S. J. Ceci, 2001) to predict 50 preschool children's performance during a field study in which they were interviewed suggestively 4 times about both a true event and a suggested event. Among the 25 children over age 4 years 6 months, tendencies on the VSSC to respond affirmatively to suggestive questions ("yield"), change answers in response to negative feedback ("shift"), and the sum of these ("total suggestibility") were all related to lack of accuracy about the true event in the field study and to both accuracy and lack of accuracy about the suggested event. Results support a 2 factor model of suggestibility. PMID- 12570099 TI - Gaze-eccentricity effects on road position and steering. AB - The effects of gaze eccentricity on the steering of an automobile were studied. Drivers performed an attention task while attempting to drive down the middle of a straight road in a simulation. Steering was biased in the direction of fixation, and deviation from the center of the road was proportional to the gaze direction until saturation at approximately 15 degrees gaze-angle from straight ahead. This effect remains when the position of the head was controlled and a reverse-steering task was used. Furthermore, the effect was not dependent on speed but reversed when the forward movement of the driver was removed from the simulation. Thus, small deviations in a driver's gaze can lead to significant impairments of the ability to drive a straight course. PMID- 12570100 TI - Anticipation skill in a real-world task: measurement, training, and transfer in tennis. AB - Anticipation skill in tennis was examined using realistic film simulations, movement-based response measures, and a portable eye movement recording system. Skilled players were faster than their less skilled counterparts in anticipating the direction of opponents' tennis strokes, with this superior performance being based, at least in part, on more effective visual search behaviors. The processes mediating superior performance were then modeled in groups of recreational tennis players using video simulation, instruction, and feedback. Players who received perceptual training improved their performance on laboratory- and field-based tests of anticipation when compared with matched placebo and control groups that did not receive any instruction regarding expert performance strategies. The approach used may have practical utility in a variety of performance contexts. PMID- 12570101 TI - Modulatory role of tachykinin NK1 receptor in cholinergic contraction of mouse trachea. AB - The role of the NK1 receptor in airway contraction induced by electrical field stimulation (EFS) was evaluated by comparing the response in NK1 receptor knockout mice (NK1R-/-) with that of NK1 receptor wild-type controls (WT). A frequency/response curve on tracheas from NK1R-/- mice and NK1R WT littermates was constructed. After incubation with [3H]choline, [3H]acetylcholine release upon EFS was measured by high-performance liquid chromatography and liquid scintillation counting. The effects of atropine (1 x 10(-6) M), tetrodotoxin (1 x 10(-6) M) and a specific NK1R antagonist (SR140333, 1 x 10(-8) M) were studied, as well as the effects of substance P (1 x 10(-5) M) on precontracted tracheas. Upon EFS, NK1R-/- mice had a significant lower trachea contractility than the NK1R WT animals, accompanied with less [3H]acetylcholine release. Pretreatment with atropine or tetrodotoxin abolished the EFS-induced contraction in both strains. Pretreatment with the NK1R antagonist SR140333 significantly reduced the contractility in the NK1R WT but not in the NK1R-/- mice. Substance P caused a small contraction in both NK1R WT and NK1R-/- mice. Substance P induced a relaxation in precontracted tracheas in NK1R WT but not in NK1R-/- mice. The data presented here provide direct evidence that the NK1 receptor augments cholinergic neurotransmission in mouse trachea. PMID- 12570102 TI - T-helper 1 cells induce alveolitis but do not lead to pulmonary fibrosis in mice. AB - T-helper (Th)1 cells have a pivotal role in the pathogenesis of hypersensitivity pneumonitis. Continued low-level exposure to the antigens may induce chronic hypersensitivity pneumonitis with lung fibrosis. Although such exposure may activate Th1 cells in the lung, it is not known whether activation of Th1 cells per se can lead to pulmonary fibrosis. To determine this, the lung pathology induced by Th1 clones was investigated. Mice (BALB/c) were injected intraperitoneally with Th1 clones 1-4 times. Each injection was performed 4 days apart and was followed by repeated exposure to aerosolised ovalbumin (OVA) once a day for 5 days. The number of macrophages and lymphocytes in bronchoalveolar lavage fluids (BALF) increased as the number of Th1 transfers increased. The number of neutrophils also increased but peaked in the second transfer and then decreased following further transfers. Increased cell infiltration, thickness of alveolar walls and number of type II cells in the lung occurred. However, histological findings showed no evidence of fibrosis and hydroxyproline levels did not increase. Findings of histology and BALF were ameliorated 2 weeks after the discontinuation of OVA exposure, indicating the reversibility of the Th1 induced pathology. In conclusion, adoptive transfer of T-helper 1 cells results in reversible alveolitis but does not lead to pulmonary fibrosis. PMID- 12570103 TI - Endogenous endothelins and nitric oxide in hypoxic pulmonary vasoconstriction. AB - The effects of endothelin receptor blockade on the pulmonary circulation have been reported variably, possibly in relation to a more or less important associated release of endogenous nitric oxide (NO). The aim of this study was to test whether endothelin antagonism would inhibit hypoxic pulmonary vasoconstriction, and if it would not, then would it do so after NO synthase inhibition. Hypoxic pulmonary vasoconstriction (HPV) was evaluated in anesthetised dogs by the increase in the mean pulmonary artery pressure (Ppa) minus occluded Ppa (Ppao) gradient in response to hypoxia (inspiratory oxygen fraction of 0.1) at constant pulmonary blood flow. Bosentan, an endothelin A and B receptor antagonist, did not affect baseline Ppa, Ppao or systemic arterial pressure (Psa) and did not alter HPV (n=8). The NO synthase inhibitor N(G)-nitro L-arginine (L-NA) did not affect baseline Ppa and Ppao, but increased Psa and enhanced HPV (n=12). The addition of bosentan in these dogs did not affect baseline Ppa or Ppao, but decreased Psa and inhibited HPV. Exhaled NO was decreased by L-NA and by bosentan and abolished by L-NA+bosentan (n=9). The authors conclude that endogenous nitric oxide is released by, and opposes the vasoconstricting effects of, endothelins in vivo, reducing systemic blood pressure and limiting hypoxic pulmonary vasoconstriction. PMID- 12570104 TI - Genetic polymorphisms associated with acute pulmonary embolism and deep venous thrombosis. AB - Frequently an inherited predisposition to thrombosis remains clinically silent until an additional environmental factor intervenes. The present study aimed to assess distribution of inherited risk factors of venous thrombosis in patients with venous thromboembolism (VTE). The prevalences of factor V Leiden (FV Leiden), prothrombin factor II G20210A (FII G20210A), C677T and A1298C of methylenetetrahydrofolate reductase (MTHFR) mutations were studied in 149 VTE patients and 100 controls. The following key risks were established: previous deep venous thrombosis or pulmonary embolism (23.5%), bed rest (34.2%), immobilisation of lower limb (10.1%), hospitalisation (30.9%) and obesity (28.9%). In 29 (19%) patients and in three (3%) controls FV Leiden was found. A significant association between VTE and FV Leiden was established. There were six (4%) carriers of the FII G20210A among VTE patients and one in the controls. No associations between VTE and MTHFR polymorphisms (C677T, A1298C) were found. In three of 149 patients both FV Leiden and FII G20210A polymorphisms were observed. The mean protein C activity was slightly, though nonsignificantly, smaller in VTE patients. In conclusion, there was a positive association between venous thromboembolism and factor V Leiden. Only a weak trend favouring a relationship between prothrombin factor II G20210A and venous thrombolism was present. No associations between common polymorphisms of methylenetetrahydrofolate reductase and venous thromboembolism were found. PMID- 12570105 TI - Single arterial occlusion to locate resistance in patients with pulmonary hypertension. AB - The purpose of this study was to determine the site of increased resistance using the arterial occlusion technique in patients with severe pulmonary hypertension. Pulmonary vascular resistance was partitioned in arterial and venous components based on double exponential fitting analysis of the pulmonary artery pressure decay curve: after balloon occlusion in 36 patients with pulmonary arterial hypertension (PAH); at baseline and during the inhalation of 20 parts per million of nitric oxide (NO); in four patients with chronic thromboembolic pulmonary hypertension; and in two patients with pulmonary veno-occlusive disease. In the patients with PAH, at baseline, mean pulmonary artery pressure was 56+/-2 mmHg (mean+/-SE), with an arterial component of resistance of 63+/-1%. Inhaled NO did not change the partition of resistance. The arterial component of resistance amounted on average to 42% and 77% in the patients with veno-occlusive disease and the patients with thromboembolic pulmonary hypertension, respectively. However, the partitioning of resistance did not discriminate between these three diagnostic categories. The occlusion technique may help to locate the predominant site of increased resistance in patients with severe pulmonary hypertension, but does not allow for a satisfactory differential diagnosis on an individual basis. PMID- 12570106 TI - Airway surface pH in subjects with cystic fibrosis. AB - The cystic fibrosis (CF) transmembrane conductance regulator protein can transport bicarbonate and may therefore regulate airway surface (AS) pH. Disturbances of AS pH could contribute to the pathophysiology of CF lung disease. Five studies were carried out including the following: study 1) nasal pH measurements were made in 25 CF and 10 non-CF adults using an antimony pH probe. Mean nasal pH was significantly lower in the CF group. Nasal potential difference may have been a confounding factor; study 2) in a fresh cohort of CF and non-CF subjects, no significant difference was found between the two groups using a gold pH probe; study 3) simultaneous nasal pH measurements were made in 15 CF and 15 non-CF adults using both probes. In the CF group, there was a trend for the antimony probe to read lower than the gold probe. In the non-CF group, the antimony probe read higher. The pH difference noted in study 1 related to technical factors; study 4) the effect of acute changes in serum acid/base balance on nasal pH was assessed in five non-CF adults. Nasal pH was not altered by either acute respiratory acidosis or alkalosis; study 5) nasal and lower airway pH was measured in five CF and six non-CF children. No difference was found between the groups. There was a correlation between nasal and lower airway pH. The authors conclude that airway surface pH does not differ between cystic fibrosis and noncystic fibrosis subjects and therefore, cystic fibrosis transmembrane conductance regulator may not play a major role in airway surface pH in vivo. PMID- 12570107 TI - Nasal nitric oxide measurements for the screening of primary ciliary dyskinesia. AB - Primary ciliary dyskinesia (PCD) presents to general practitioners with symptoms pertinent to a variety of specialists because of the involvement of ciliated epithelium in the upper/lower respiratory tract, ears, eyes and genital tract. There is no easy, reliable screening test for PCD, and thus, the majority of patients remain undiagnosed. Nitric oxide (NO) is measurable in nasal air of normal subjects and found to be low in cystic fibrosis (CF) and very low in PCD. Recently, it was suggested to play an important role in regulating ciliary motility. The aim of this study was to evaluate whether measurements of nasal NO could be used to screen for PCD. Nasal NO was measured from the nasal cavity by a chemiluminescence analyser in subjects with PCD, healthy controls, CF, idiopathic bronchiectasis, Young's syndrome and lone sinusitis. Nasal NO was significantly lower in PCD (64.0+/-36.6) compared with normal controls (759+/-145.8), idiopathic bronchiectasis (734+/-163.7), CF (447.5+/-162.6), lone sinusitis (1487+/-734) and Young's syndrome (644+/-129.9). Nasal NO was also significantly lower in PCD than CF patients. Measurement of nasal nitric oxide may therefore be used clinically in various specialities to screen suspected patients for primary ciliary dyskinesia. PMID- 12570108 TI - Effect of oxygen on breath markers of oxidative stress. AB - Supplemental oxygen is often administered to induce hyperoxia in nonhypoxic patients for indications such as chest pain, despite lack of evidence of clinical benefit. Induced hyperoxia is potentially toxic, since it may increase oxidative stress and peroxidative damage to deoxyribonucleic acid, lipids and proteins. The aim of this study was to establish whether supplemental oxygen induces oxidative stress in nonhypoxic subjects. Breath markers of oxidative stress were measured in 31 healthy subjects before and after breathing 28% oxygen at 2.0 L x min(-1) via nasal prongs for 30 min while resting. The criterion standard of oxidative stress was the breath methylated alkane contour (BMAC), a three-dimensional plot of the alveolar gradients of C4-C20 alkanes and monomethylated alkanes produced by lipid peroxidation. Volatile organic compounds (VOCs) in breath were assayed by gas chromatography and mass spectroscopy, and the BMACs before and after oxygenation were compared. Following oxygenation, there was a significant increase in mean volume under the curve of the BMAC and in alveolar gradients of three VOCs: 3-methyltridecane, 3-methylundecane and 5-methylnonane. Breath markers of oxidative stress were significantly increased in normal volunteers breathing supplemental oxygen for 30 min. PMID- 12570109 TI - Markers of activity in clinically recovered human leukocyte antigen-DR17-positive sarcoidosis patients. AB - Scandinavian human leukocyte antigen-DR17-positive (DR17+) sarcoidosis patients are characterised by a good prognosis. They also reveal an accumulation in bronchoalveolar lavage fluid of T-lymphocytes expressing the T-cell receptor V gene segment AV2S3 at disease onset. The authors of this study wished to establish whether AV2S3 T-lymphocyte accumulation changes from disease onset to clinically resolved disease and how this relates to other activity parameters. Bronchoalveolar lavage fluid and serum from nine DR17+ sarcoidosis patients were examined at disease onset and after spontaneous resolution of clinical and radiographical signs of disease. Nine DR17+ patients with lung accumulated CD4+ AV2S3+ T-cells were investigated after clinically recovery. At re-examination the percentage of CD4+ AV2S3+ lymphocytes in bronchoalveolar lavage fluid was normalised (29 versus 5.4%). A significant reduction in lymphocyte percentage (14 versus 4.4%) and a decrease in cellular concentration (179x10(6) x L(-1) versus 111x10(6) x L(-1)) and CD4/CD8 ratio (5.2 versus 2.4) were also seen. In serum, the activity of angiotensin-converting enzyme (24.9 versus 14.0 U x mL(-1)) as well as the levels of neopterin (7.8 versus 5.3 nmol x L(-1)) decreased significantly after recovery. These results indicate that the locally accumulated AV2S3 positive T-lymphocytes in bronchoalveolar lavage are involved in the pathogenic process of sarcoidosis in this patient group. PMID- 12570110 TI - Home hospitalisation of exacerbated chronic obstructive pulmonary disease patients. AB - It was postulated that home hospitalisation (HH) of selected chronic obstructive pulmonary disease (COPD) exacerbations admitted at the emergency room (ER) could facilitate a better outcome than conventional hospitalisation. To this end, 222 COPD patients (3.2% female; 71+/-10 yrs (mean+/-SD)) were randomly assigned to HH (n=121) or conventional care (n=101). During HH, integrated care was delivered by a specialised nurse with the patient's free-phone access to the nurse ensured for an 8-week follow-up period. Mortality (HH: 4.1%; controls: 6.9%) and hospital readmissions (HH: 0.24+/-0.57 controls: 0.38+/-0.70) were similar in both groups. However, at the end of the follow-up period, HH patients showed: 1) a lower rate of ER visits (0.13+/-0.43 versus 0.31+/-0.62); and 2) a noticeable improvement of quality of life (delta St George's Respiratory Questionnaire (SGRQ), -6.9 versus 2.4). Furthermore, a higher percentage of patients had a better knowledge of the disease (58% versus 27%), a better self-management of their condition (81% versus 48%), and the patient's satisfaction was greater. The average overall direct cost per HH patient was 62% of the costs of conventional care, essentially due to fewer days of inpatient hospitalisation (1.7+/-2.3 versus 4.2+/-4.1 days). A comprehensive home care intervention in selected chronic obstructive pulmonary disease exacerbations appears as cost effective. The home hospitalisation intervention generates better outcomes at lower costs than conventional care. PMID- 12570111 TI - Disease severity and the effect of fluticasone propionate on chronic obstructive pulmonary disease exacerbations. AB - Exacerbations of chronic obstructive pulmonary disease (COPD) are associated with worse health and increased healthcare utilisation. The Inhaled Steroids in Obstructive Lung Disease in Europe (ISOLDE) study in COPD showed a 26% reduction in the yearly rate of exacerbations in patients treated with fluticasone propionate (FP) compared to placebo, but did not indicate which patients showed greatest benefit. In this study the patients were stratified into mild and moderate-to-severe COPD using the American Thoracic Society criterion of forced expiratory volume in one second (FEV1) 50% predicted, and the total number of exacerbations and those requiring treatment with oral corticosteroids were examined. There were 391 (195 FP) patients with mild COPD and 359 (180 FP) patients with moderate-to-severe disease. The exacerbation rate was highly skewed in mild disease, but more normally distributed in moderate-to-severe disease. FP reduced the overall exacerbation rate in moderate-to-severe disease (FP median rate 1.47 yr(-1), placebo 1.75 yr(-1)), but not in mild disease (FP 0.67 yr(-1), placebo 0.92 yr(-1)). FP use was associated with fewer patients with > or = 1 exacerbation x yr(-1) being treated with oral corticosteroids (mild: FP 8%, placebo 16%; moderate-to-severe: FP 17%, placebo 30%). Effects of fluticasone propionate on exacerbations were seen predominantly in patients with a postbronchodilator forced expiratory volume in one second <50% predicted. These data support recommendations in the Global Initiative for Chronic Obstructive Disease treatment guidelines that inhaled corticosteroids should be considered in patients with moderate-to-severe chronic obstructive pulmonary disease who experience recurrent exacerbations. PMID- 12570112 TI - Efficacy and safety of budesonide/formoterol in the management of chronic obstructive pulmonary disease. AB - The efficacy and safety of budesonide/formoterol in a single inhaler compared with placebo, budesonide and formoterol were evaluated in patients with moderate to-severe chronic obstructive pulmonary disease (COPD). In a 12-month, randomised, double-blind, placebo-controlled, parallel-group study in 812 adults (mean age 64 yrs, mean forced expiratory volume in one second (FEV1) 36% predicted normal), patients received two inhalations twice daily of either budesonide/formoterol (Symbicort) 160/4.5 microg (delivered dose), budesonide 200 microg (metered dose), formoterol 4.5 microg or placebo. Severe exacerbations and FEV1 (primary variables), peak expiratory flow (PEF), COPD symptoms, health related quality of life (HRQL), mild exacerbations, use of reliever beta2-agonist and safety variables were recorded. Budesonide/formoterol reduced the mean number of severe exacerbations per patient per year by 24% versus placebo and 23% versus formoterol. FEV1 increased by 15% versus placebo and 9% versus budesonide. Morning PEF improved significantly on day 1 versus placebo and budesonide; after 1 week, morning PEF was improved versus placebo, budesonide and formoterol. Improvements in morning and evening PEF versus comparators were maintained over 12 months. Budesonide/formoterol decreased all symptom scores and use of reliever beta2-agonists significantly versus placebo and budesonide, and improved HRQL versus placebo. All treatments were well tolerated. These results suggest a role for budesonide/formoterol in the long-term management of moderate-to-severe chronic obstructive pulmonary disease. PMID- 12570113 TI - Bronchodilation test in COPD: effect of inspiratory manoeuvre preceding forced expiration. AB - The effects of an inspiratory manoeuvre preceding forced expiration on functional tests performed under routine conditions before and after inhalation of a bronchodilator drug (salbutamol) were assessed on 150 consecutive chronic obstructive pulmonary disease outpatients. The patients performed forced vital capacity manoeuvres either immediately after a rapid inspiration (manoeuvre no. 1) or after a slow inspiration with a 4-6 s pause (manoeuvre no. 2). Under baseline conditions, forced expiratory volume in one second (FEV1) values were 8% (% control) larger with manoeuvre no. 1 than no. 2. FEV1 values increased with salbutamol administration by approximately 8% and were, on average, still 7% larger with manoeuvre no. 1 than no. 2. The incidence of reversibility, assessed according to American Thoracic Society criteria, was 76% when manoeuvre no. 2 was selected to represent baseline conditions and manoeuvre no. 1 was chosen to represent the effects of bronchodilator administration, whereas the lowest incidence (2%) was found when manoeuvre no. 1 was selected to represent baseline conditions and manoeuvre no. 2 was chosen to represent the effects of bronchodilator administration. These results indicate that the time dependence of the forced vital capacity manoeuvre has an important impact on the assessment of routine lung function in a clinical setting and supports the notion that the time course of the inspiration preceding the forced vital capacity manoeuvre should be standardised. PMID- 12570114 TI - Effect of inhaled bronchodilators on inspiratory capacity and dyspnoea at rest in COPD. AB - It has been shown that patients with chronic obstructive pulmonary disease (COPD) develop dynamic hyperinflation (DH), which contributes to dyspnoea and exercise intolerance. Formoterol, salmeterol and oxitropium have been recommended for maintenance therapy in COPD patients, but their effect on DH has only been assessed for salmeterol. The aim of the present study was to compare the acute effect of four inhaled bronchodilators (salbutamol, formoterol, salmeterol and oxitropium) and placebo on forced expiratory volume in one second, inspiratory capacity, forced vital capacity and dyspnoea in COPD patients. A cross-over, randomised, double-blind, placebo-controlled study was carried out on 20 COPD patients. Patients underwent pulmonary function testing and dyspnoea evaluation, in basal condition and 5, 15, 30, 60 and 120 min after bronchodilator or placebo administration. The results indicate that in chronic obstructive pulmonary disease patients with decreased baseline inspiratory capacity, there was a much greater increase of inspiratory capacity after bronchodilator administration, which correlated closely with the improvement of dyspnoea sensation at rest. For all bronchodilators used, inspiratory capacity reversibility should be tested at 30 min following the bronchodilator. On average, formoterol elicited the greatest increase in inspiratory capacity than the other bronchodilators used, though the difference was significant only with salmeterol and oxitropium. The potential advantage of formoterol needs to be tested in a larger patient population. PMID- 12570115 TI - The role of concomitant respiratory diseases on the rate of decline in FEV1 among adult asthmatics. AB - Several recent reports have presented evidence suggesting that adults with asthma have an accelerated rate of decline in pulmonary function compared with nonasthmatics. However, most of these studies have not taken into account the possible effect of comorbid lung disease in addition to asthma. This study was designed to determine if comorbid respiratory diseases modify or otherwise change the estimates of decline in forced expiratory volume in one second (FEV1). Study subjects were White, non-Mexican, American participants, who were > or = 20 yrs of age at the initial survey and had at least one pulmonary function testing. Respiratory disease status, based on self-reported questionnaires and pulmonary function tests, were assessed during 12 surveys spanning a period of up to 20 yrs. There were 2,926 subjects who met the inclusion criteria, 214 (7.3%) had physician-confirmed asthma, 325 (11.1%) chronic obstructive pulmonary disease (COPD), and 125 (4.3%) both physician-confirmed asthma and COPD. Longitudinal analysis revealed that among those with longstanding asthma, FEV1 values were low but did not decline at a more rapid rate than normal. Likewise, subjects with both asthma and COPD had the lowest levels of FEV1, but also a rate of decline that was not significantly increased. Only those with COPD in the absence of asthma experienced both a low initial FEV1 level and a significantly steeper rate of decline. Based on these findings, the authors conclude that forced expiratory volume in one second does not decline more rapidly in asthmatics or in those with asthma and chronic obstructive pulmonary disease, compared with nonasthmatics. PMID- 12570116 TI - Use of anti-inflammatory therapy and asthma mortality in Japan. AB - Asthma treatment guidelines were introduced in Japan in the 1990s, insisting as elsewhere, on the importance of anti-inflammatory therapy. The present study assessed whether use of anti-inflammatory medications was associated with a decrease in asthma mortality in Japan, the first country to use leukotriene receptor antagonists. A population-based ecological study was conducted, spanning the period 1987-1999, among people aged 5-34 yrs in Japan. The association between the yearly rate of asthma death and sales of inhaled corticosteroids and leukotriene receptor antagonists was estimated using Poisson regression. The yearly asthma death rate was stable at 6-7 deaths per million before the introduction of leukotriene receptor antagonists in 1995 and decreased by 23% thereafter, reaching 3.5 per million in 1999. The rate of asthma death was found to decrease with increasing use of both leukotriene receptor antagonists and inhaled corticosteroids. The rate ratio of asthma death was 0.96 per 1 million 25 day treatment courses of inhaled corticosteroids and 0.80 for every 1 million 25 day treatment courses of leukotriene receptor antagonists, consumed per year in Japan. The increasing use of inhaled corticosteroids and leukotriene receptor antagonists may have contributed to the significant reduction in asthma mortality among young asthmatics in Japan. PMID- 12570117 TI - Socioeconomical conditions as risk factors for bronchial asthma in children aged 4-5 yrs. AB - A number of factors are associated with the development of childhood asthma. The purpose of this study was to establish the prevalence of childhood asthma and to explore the socioeconomic background factors associated with childhood asthma in a Norwegian cohort of children aged 4-5 yrs. A questionnaire was given to parents in connection with the ordinary child control of 4-5-yr-old children in Vestfold county, Norway. In addition to the question "Has the child at present or ever had asthma?", a number of medical and socioeconomical background factors were registered. Of the 2,430 parents, 1,913 (79%) responded. Of the 163 (cumulative prevalence 8.7%) children with confirmed asthma, 19 did not use any medication and were regarded as having outgrown their asthma. Several background factors were significantly associated with asthma in a logistic regression analysis: few rooms at home, psychosocial problems, fever more than three times during the last year, hay fever, reaction to food and mother or father with chronic disease. The findings indicate that socioeconomic background factors are associated with asthma in childhood, in addition to other known risk factors. PMID- 12570118 TI - Evaluation of an individualised asthma programme directed at behavioural change. AB - An individualised asthma programme directed at behavioural change was evaluated in asthmatic subjects who reported complaints and impairment, despite adequate medical treatment. Mild-to-moderate asthma patients (n=23) were randomly assigned to a programme or waiting list condition. Outcome measures were: McMaster Asthma Quality of Life Questionnaire, Asthma Symptom Checklist, Negative Emotionality Scale, Knowledge, Attitude and Self-Efficacy Asthma Questionnaire, Adherence Scale, and peak flow measurements. Both groups were evaluated at three consecutive moments, each separated by 3 months; the programme was delivered between the first two evaluations. At onset the patient received a workbook containing information, exercises and homework assignments. Psycho-education, behavioural and cognitive techniques were introduced during six 1-h individual sessions. Compared with controls the programme group reported less symptoms (obstruction, fatigue), better quality of life (activity, symptoms, emotions), decreased negative affectivity, and increased adherence, immediately after finishing the programme and at 3 months follow-up. All three cognitive variables (knowledge, attitude towards asthma, self-efficacy) and day and night peak flow ratings improved in the programme group but not in the waiting list group. Participation in an individualised programme resulted in improvement of asthma morbidity, and asthma-related behaviour and cognitions, in subjects reporting symptoms and impairment despite adequate medical therapy. PMID- 12570119 TI - Cost of illness of atopic asthma and seasonal allergic rhinitis in Germany: 1-yr retrospective study. AB - The purpose of this study was to evaluate the cost of illness of moderate-to severe atopic asthma and/or seasonal allergic rhinitis (SAR) in Germany from the perspective of third-party payers (TPP) and patients. Five-hundred patients (276 children/adolescents) with moderate-to-severe asthma and/or SAR were included in this cross-sectional study. Information was collected using a specific patient questionnaire and the abstraction of patient records. Overall, annual costs per patient increased with the severity of atopic asthma and if it was associated with SAR. The average annual cost of SAR was Euro1,089 per child/adolescent and Euro1,543 per adult. Annual costs of severe asthma plus SAR increased to Euro7,928 per child/adolescent and to Euro9,287 per adult. For TPPs, the main cost drivers were medication, hospitalisation, and rehabilitation. The most significant costs for patients were household modifications. For children/adolescents, 60-78% of the expenditures were direct costs, while in adults, 58% of expenditures were indirect costs. It was also observed that patients with moderate and severe asthma used inhaled corticosteroids less frequently than recommended by treatment guidelines. In summary, the total cost for patients increases with the severity of atopic asthma and/or seasonal allergic rhinitis and indirect costs represent a large proportion of the total cost. PMID- 12570120 TI - Distribution of therapeutic response in asthma control between oral montelukast and inhaled beclomethasone. AB - The distribution of responses in study populations provides a novel method of comparing the benefit of two treatments. This 6-week, randomised, placebo controlled, double-blind study compared the effectiveness of oral montelukast with inhaled beclomethasone in chronic asthma by assessing the distribution and overlap of patient responses to therapy, as measured by a clinical outcome (asthma control days). A total of 730 adult patients with asthma, age 15-65 yrs, with a forced expiratory volume in one second (FEV1) at baseline of 50-85% of predicted and > or = 15% improvement in FEV1 after inhaled beta-agonist were enrolled. After a 2-week placebo run-in period, patients were randomly allocated to receive montelukast (10 mg once daily), inhaled beclomethasone (200 microg twice daily) or placebo. The primary end-point (per cent of asthma control days) was compared between treatments as the overlap in the response distributions. The overlap of the distribution of responses between the montelukast and beclomethasone groups was 89% for per cent asthma control days and 96% for change from baseline in FEV1. The mean (+/-SD) per cent asthma control days in the montelukast and beclomethasone groups was significantly higher than that in the placebo group (placebo 40.0+/-35.8, montelukast 50.7+/-37.1, beclomethasone 57.9+/-36.1). The mean differences between montelukast and placebo, beclomethasone and placebo, and montelukast and beclomethasone were significant. The mean per cent change (+/-SD) from baseline in FEV1 was 12.1+/-18.7 and 13.9+/ 20.8 in the montelukast and beclomethasone groups, respectively, and significantly greater than that in the placebo group (6.4+/-20.1); there was no significant difference between the montelukast and beclomethasone groups in mean values or response distribution. There was also no difference among treatment groups in the frequency of adverse experiences. A comparison of the response distribution is an important approach to comparing therapies; montelukast and beclomethasone provided similar response distributions for the end-point of per cent asthma control days over a 6-week treatment period. PMID- 12570121 TI - Monitoring the quality of laboratories and the prevalence of resistance to antituberculosis drugs: Italy, 1998-2000. AB - In 1998 a network of 20 regional tuberculosis (TB) laboratories (the Italian Multicentre Study on Resistance to Antituberculosis drugs (SMIRA) network) was established in Italy to implement proficiency testing and to monitor the prevalence of drug resistance nationwide. The network managed 30% of all TB cases reported in Italy each year. The aim of the present report is to describe: 1) the accuracy of drug-susceptibility testing in the network; 2) the prevalence of drug resistance for the period 1998-2000. Data were collected from the network laboratories. Sensitivity to streptomycin and ethambutol increased from the first survey (1998-1999) to the second survey (2000) from 87.7 to 91.9%. Specificity, predictive values for resistance and susceptibility, efficiency and reproducibility were consistent in both surveys. In previously untreated cases, the prevalence of multidrug-resistance was the same in both surveys (1.2%), while a slight decrease from the first to the second survey was observed for monoresistance to rifampicin (from 0.8 to 0.4%) and isoniazid (from 2.9 to 2%). The significant association found between isoniazid resistance and immigration is a useful indicator for both clinicians managing individual tuberculosis cases and public health services planning control strategies. PMID- 12570122 TI - Effectiveness of oral moxifloxacin in standard first-line therapy in community acquired pneumonia. AB - Based on recent guidelines for the management of community-acquired pneumonia, this study was designed to evaluate the effectiveness of a new fluoroquinolone compared with standard antimicrobial regimens, in conditions relating as closely as possible to the real world setting. In this study, 564 patients were randomised to either oral moxifloxacin (400 mg o.d.) or to standard oral therapy (amoxicillin 1 g t.i.d. or clarithromycin 500 mg b.i.d. alone or in combination) for up to 14 days using a double-blind procedure. The choice between the three standard regimens was made by the clinician prior to randomisation. Clinical response, quality of life, symptoms, healthcare resources and safety were assessed. In the per-protocol population, clinical success was reported for 201 of 215 (93.5%) and 217 of 231 (93.9%) in the moxifloxacin and standard groups, respectively, at 7-10 days post-therapy. At 28-35 days follow-up, continued clinical cure was observed in 183 of 192 (95.3%) moxifloxacin and 207 of 221 (93.7%) standard groups. Drug-related adverse events were reported in 55 of 274 (20%) moxifloxacin and 86 of 279 (31%) standard patients with diarrhoea >5%. Oral moxifloxacin monotherapy was as effective as, and better tolerated than, optimal antibiotic strategy represented either by mono- or combination therapy (amoxicillin and/or clarithromycin) in community-acquired pneumonia management. PMID- 12570123 TI - Bronchoalveolar lavage fluid findings in children with hypersensitivity pneumonitis. AB - Bronchoalveolar lavage (BAL) has been shown to be useful in the diagnosis of hypersensitivity pneumonitis (HP) in adults, the typical constellation being lymphocytosis with a decrease in the CD4/CD8 ratio. Only limited data exist for the diagnostic value of BAL cytology in paediatric patients with this disorder. Children aged 6-15 yrs (n=9) with acute HP were studied. BAL was performed before initiation of anti-inflammatory treatment via a flexible bronchoscope in the middle lobe with 3 mL x kg body weight(-1) normal saline warmed to body temperature; BAL cytology and lymphocyte surface markers were compared with age matched controls. The percentage of lymphocytes was significantly increased in all patients with HP. No significant differences were observed in the CD4/CD8 ratio between children without lung disease and those with HP. Increased expression of human leukocyte antigen-DR was found in seven of eight children with HP, whereas natural killer cells were elevated in five of eight children. Every patient had at least one of these two alterations in BAL fluid in addition to lymphocytosis. It was concluded that while lymphocytosis is generally present in children with hypersensitivity pneumonitis, the CD4/CD8 ratio is not increased in these patients. Assessing natural killer cells and human leukocyte antigen-DR expression appears to be a helpful adjunct in the diagnosis of paediatric patients with this disorder. PMID- 12570124 TI - Urinary leukotriene E4 in preschool children with acute clinical viral wheeze. AB - Cysteinyl leukotrienes (cystLTs) are important mediators of wheeze in atopic asthma, but the role of cystLTs in the pathogenesis of preschool viral wheeze (PVW) is unclear. Therefore, evidence for increased production of cystLTs in PVW was sought. Urinary leukotriene E4 (uLTE4) and serum total immunoglobulin (Ig)E were measured in children (1-5 yrs) with PVW during an acute attack (n=44) and in the convalescent phase (n=19), and compared with normal controls (n=15). The effect of atopic sensitisation was assessed in a separate group of atopic controls (n=6) in whom only uLTE4 was measured. The levels of uLTE4 were similar in normal and atopic controls and increased in acute PVW (median (interquartile range) 165 (101-285) versus 125 (82-163) ng x mM creatinine(-1)). Stratification by IgE showed that whereas uLTE4 was increased in 23 children with acute PVW and IgE > 95th percentile (median 211 (118-312) ng x mM creatinine(-1)), uLTE4 was not increased in the 21 children with acute PVW and IgE < or = 95th percentile. In the convalescent phase, uLTE4 fell in the subgroup with high IgE but not in the subgroup with low IgE. It is concluded that increased cysteinyl leukotriene production during acute preschool viral wheeze is associated with high serum immunoglobulin E. PMID- 12570125 TI - Pulmonary arterial hypertension in children. AB - For physicians to admit that a group of patients remains for whom no cure is available in modern medicine is intellectually unsatisfying. Pulmonary arterial hypertension is a rare condition. Because the symptoms are nonspecific and the physical finding can be subtle, the disease is often diagnosed in its later stages. The natural history of pulmonary arterial hypertension is usually progressive and fatal. At the 1998 Primary Pulmonary Hypertension World Symposium, clinical scientists from around the world gathered to review and discuss the future of pulmonary arterial hypertension. Bringing together experts from a variety of disciplines provided the opportunity for a better understanding of the pathology, pathobiology, risk factors, genetics, diagnosis and treatment for pulmonary arterial hypertension. Remarkable progress has been made in the field of pulmonary arterial hypertension over the past several decades. The pathology is now better defined and significant advances have occurred in understanding the pathobiological mechanisms. Risk factors have been identified and the genetics have been characterised. Advances in technology allow earlier diagnosis as well as better assessment of disease severity. Therapeutic modalities such as new drugs, e.g. epoprostenol, treprostinil and bosentan, and surgical interventions, e.g. transplantation and blade septostomy, which were unavailable several decades ago, have had a significant impact on prognosis and outcome. Thus, despite the inability to really cure pulmonary arterial hypertension, therapeutic advances over the past two decades have resulted in significant improvements in the outcome for children with various forms of pulmonary arterial hypertension. This review of pulmonary arterial hypertension will highlight the key features of pulmonary hypertension in infants and children and the current understanding of pulmonary arterial hypertension with specific recommendations for current practice and future directions. PMID- 12570126 TI - Biomarkers of lipid peroxidation, airway inflammation and asthma. AB - Oxidative stress, specifically lipid peroxidation, is believed to contribute to the pathophysiology of asthma. This review highlights the pathways through which reactive oxygen species (ROS) may lead to lipid peroxidation. The potential of both the innate and acquired immune systems to activate inflammatory cells and release ROS that may overwhelm the host antioxidant defences and cause lipid peroxidation, accompanied by detrimental pathophysiological effects, are discussed. Despite the evidence demonstrating the importance of lipid peroxidation, systematic characterisation of oxidative stress and antioxidant defences has not been undertaken, largely due to the lack of appropriate biomarkers. This review discusses the emergence of isoprostanes (specifically 8 iso-prostaglandin F2alpha) as reliable, in vivo markers of lipid peroxidation, which provides an appropriate tool for studying oxidative stress. Furthermore, the development of techniques to study induced sputum and breath condensate, derived directly from the airway surface, enables the site of oxidative damage to be closely assessed. Evidence suggests that dietary changes that have occurred over recent years have increased susceptibility to lipid peroxidation, due to reduced antioxidant defences. To date, the limited number of long-term (>1 week) supplementation trials have been promising. However, the development of techniques to study isoprostanes in airway-lining fluid pave the way for further studies investigating the potential for antioxidant supplements to be used as routine therapy in asthma. PMID- 12570127 TI - Acute interstitial pneumonia: report of a series. AB - Four cases of acute interstitial pneumonia (AIP) are described with special emphasis on clinical background, lung imaging and bronchoalveolar lavage findings. A retrospective chart review of four patients with histologically proven AIP, diagnosed between 1998 and 2000, was carried out. Clinical data, bronchoalveolar lavage (BAL) findings, high-resolution computed tomography (HRCT) and histological features were analysed. Three patients died and only one is in follow-up. HRCT showed areas of ground glass attenuation and alveolar consolidation in all patients. Histology, documented by open lung biopsy or autopsy specimens, was consistent with the organising form of diffuse alveolar damage pattern. BAL findings were characteristic, with a huge neutrophilia associated with scattered atypical type II pneumocytes collected in clusters with extracellular amorphous material (fragments of hyaline membranes) observed in two out of three cases. In this paper, four cases of acute interstitial pneumonia are reported in detail. The poor prognosis associated with this entity has been confirmed and the possible diagnostic role of the bronchoalveolar lavage is emphasised. PMID- 12570128 TI - Epoprostenol therapy for primary pulmonary hypertension after rejection of a single donor lung. AB - Before prostacyclins became available, lung or heart/lung transplantation was the only effective treatment for patients with primary pulmonary hypertension (PPH) who deteriorated under supportive medical treatment. Unfortunately, acute and chronic rejections occur in a large number of cases, limiting the average survival to 4.5 yrs. A female patient, age 35 yrs, was diagnosed with PPH and underwent single lung transplantation. Despite aggressive immunosuppressive therapy, the patient had several episodes of acute rejection. Eventually, chronic rejection with bronchiolitis obliterans developed. After 5 yrs, the donor lung was no longer functional. The patient was in New York Heart Association (NYHA) class 4, had a 6-min walking distance of 50 m and a resting arterial oxygen tension (Pa,O2) of 9.8 kPa (74 mmHg) when using 3 L x min(-1) of oxygen. Epoprostenol treatment was started and the patient showed remarkable improvement. After 17 months the patient was NYHA class 2, walked 503 m in 6 min and had a resting Pa,O2 of 10.9 kPa (82 mmHg) without supplemental oxygen. In this patient, treatment with epoprostenol was effective after rejection of a single donor lung transplanted for primary pulmonary hypertension. PMID- 12570129 TI - Exhaled breath temperature in asthma. PMID- 12570130 TI - Exhaled carbon monoxide in lung disease. PMID- 12570131 TI - Intraoperative imaging in neurosurgery. MRI, CT, ultrasound. Introduction. PMID- 12570132 TI - Future perspectives in intraoperative imaging. AB - Of all the advances in imaging science in the past twenty years, none has had a greater impact than Magnetic Resonance Imaging. Since its introduction as a diagnostic tool in the mid-1980's, MRI has evolved into the premier neuroimaging modality, and with the addition of higher field magnets, we are able to achieve spatial resolution of such superb quality that even the most exquisite details of the brain anatomy can be visualized. With the implementation of intraoperative, neurosurgical MRI, we can not only monitor brain shifts and deformations; we can achieve intraoperative navigation using intraoperative image updates. In the future, intraoperative MRI can be used not only to localize, target, and resect brain tumors and other lesions but also to fully comprehend the surrounding cortical and white matter functional anatomy. In addition to the inclusion of new imaging methods such as diffusion tensor imaging, new therapeutic methods will be applied. Especially encouraging are the promising results in MRI-guided Focused Ultrasound Surgery, in which the non-invasive thermal ablation of tumors is monitored and controlled by MRI. With the clinical introduction of these advances, intraoperative MRI is changing the face of Neurosurgery today. PMID- 12570133 TI - How to choose a magnet--reflections on the development of MRI-guided neurosurgery. PMID- 12570134 TI - Intraoperative MR at 1.5 Tesla--experience and future directions. AB - The objective of this report is to present and contrast the development of the different intraoperative MR systems that are currently in use. The manuscript focuses on the design and clinical experience of a 1.5 Tesla MR system, based on a movable magnet. This configuration is similar to the operating microscope and other surgical adjuncts, with MR technology moved to and from the patient as needed. The system has been used to monitor 294 neurosurgical procedures. including CNS neoplasia. epilepsy, cervical spine disorders, arteriovenous malformations, cavernomas and aneurysms. In many cases the surgical procedure was significantly altered by intraoperatively acquired MRI. Future developments include the construction of a 3 Tesla intraoperative MR system and an ambidextrous MR-compatible robot. This seamless integration of robotic technology into an intraoperative MR environment may well revolutionize neurosurgery. PMID- 12570135 TI - Influence of 1.5-Tesla intraoperative MR imaging on surgical decision making. AB - To determine the frequency that high-field magnetic resonance (MR) imaging sequences influenced surgical decision making during intraoperative MR-guided surgery. From January 1997 to February 2001, 346 MR-guided procedures were performed using a 1.5-Tesla MR system (NT-ACS, Philips Medical Systems). This system can perform functional MR imaging (fMRI), diffusion weighted imaging (DWI), MR spectroscopy (MRS), MR angiography (MRA), and MR venography (MRV) in addition to T1-weighted, T2-weighted, and turbo FLAIR (fluid-attenuated inversion recovery) imaging. FMRI was used to determine areas of brain activation for language, motor function, and memory. DWI was utilized after tumor resection to exclude cerebral ischemia or infarction. MRS was obtained to identify areas of elevated choline that were suspected to correlate with tumor presence. MRA and MRV localized vascular structures adjacent to tumors prior to resection. The intraoperative procedures performed included 140 brain biopsies of which 82 utilized a trajectory guide and prospective stereotaxy. MRS was used in 42 biopsies (30%), of which 29 had turbo spectroscopic imaging (TSI) and 21 had single voxel spectroscopy (SVS). In all biopsy cases, diagnostic tissue was obtained. There were 103 tumor resections of which 18 (17%) had MRS. Functional MRI was used in 17 cases; 3 biopsies (2%) and 14 planned resections (14%). Speech function was localized in 3 cases, memory function in 3, and motor function in 11. In one case where the motor function of the tongue was intimately involved with a low-grade glioma, resection was not attempted. DWI was used in less than 10% of tumor resections. MRA and MRV were performed in 3 (3%) and 2 (2%) of tumor resections, respectively. The imaging capabilities (i.e., fMRI, DWI, MRA, MRV) associated with high-field intraoperative MR influenced surgical decision making primarily for tumor resections. MRS influenced target selection during brain biopsy. PMID- 12570136 TI - Tumor resection in a shared-resource magnetic resonance operating room: experience at the University of Cincinnati. AB - INTRODUCTION: At the University of Cincinnati, we have developed a shared resource magnetic resonance operating suite that facilitates performance of both neurosurgical and diagnostic procedures in a single unit. METHODS: The shared resource magnetic resonance operating suite utilizes a Hitachi AIRIS II, 0.3-T, vertical field, open MRI unit located in the MROR. This magnet can be used for both diagnostic and interventional procedures. The addition of a rotating operating table permits neurosurgical procedures to be performed outside of the 5 G line using standard neurosurgical equipment and operating microscopes. RESULTS: We review our results with the shared-resource magnetic resonance operating room including the tabulated results from 30 transsphenoidal procedures and 63 glioma procedures. In addition, 2832 diagnostic procedures have been performed in the first 4 years of use. CONCLUSION: The shared-resource intraoperative MRI facility produces high-quality intraoperative imaging studies, equal to those of high resolution magnets, and is valuable in enabling the surgeon to achieve the planned degree of resection of glioma and pituitary tumors. The ability to perform diagnostic procedures in a shared unit has been a cost-effective solution for our institution. PMID- 12570137 TI - Surgical management of intracranial gliomas--does radical resection improve outcome? AB - This paper discusses the arguments for and against radical tumor resection as a strategy for treatment of cerebral gliomas. METHOD: Data from the Glioma Outcome Project were analyzed to determine whether survival could be related to extent of resection in 666 patients treated by biopsy or resection for malignant cerebral gliomas. FINDINGS: Consistent survival advantages were noted for those patients treated with resection. INTERPRETATION: This observational study, although undoubtedly reflecting selection bias, provides data that support resection as a major factor in survival after surgery for malignant gliomas. PMID- 12570138 TI - Glioma surgery evaluated by intraoperative low-field magnetic resonance imaging. AB - OBJECTIVE: To give an overview on intraoperative magnetic resonance (MR) imaging in glioma surgery. MATERIAL AND METHODS: MR imaging was performed using a 0.2T scanner, located in a radiofrequency-shielded operating theatre. Two setups were used: surgery either in a neighbouring operating theatre, or directly at the 5G line. Additionally, in gliomas adjacent to eloquent brain areas microscope- or pointer-based neuronavigation with integrated functional data was applied. 106 gliomas were among the 330 patients investigated in the last 5 years. RESULTS: We did not observe complications attributable to intraoperative MR imaging. Image quality was sufficient to evaluate the extent of the tumour resection in the majority of cases. Intraoperative imaging revealed remaining tumour in 63%. In a total of 26% patients further tumour could be removed due to the results of intraoperative imaging, increasing the rate of complete tumour removal especially in the low-grade tumours. The additional use of functional neuronavigation prevented an increased morbidity. CONCLUSION: Intraoperative MR imaging offers the possibility of further tumour removal during the same surgical procedure in case of tumour remnants, increasing the rate of complete tumour removal. The effects of brain shift can be compensated for using intraoperative image data for updating. PMID- 12570139 TI - Clinical results in MR-guided therapy for malignant gliomas. AB - The prognostic impact of the extent of tumour resection in surgery of malignant glioma patients remains controversial. We report the results of cumulative survival of malignant glioma patients operated with MR-guidance. Patients with complete tumour removal were compared with a population of patients with incomplete tumour removal. A 0.5 T scanner was used to criticize the extent of resection during surgery. In total no significant difference could be found, however there is a tendency that complete tumour removal seems to be associated with a slightly increased median survival time. PMID- 12570140 TI - Intraoperative MRI for pediatric tumor management. AB - The emergence of intraoperative MRI has opened new doors for the surgical treatment of pediatric disorders. This technology will hopefully not only improve the surgeon's ability to obtain complete tumor resections with minimal damage to surrounding structures, but also allows surgeons to perform various procedures via less invasive measures. We performed a total of 38 procedures in 36 children in our intraoperative MRI system (GE Signa SP, open configuration). All procedures were performed within the magnet bore, which allows for either continuous real-time or periodic imaging. Procedures included craniotomy for tumor resection, open biopsy, stereotactic biopsy or catheter placement into a tumor-related cyst. There were no infectious, hemorrhagic or neurological complications. Intraoperative MRI is an useful tool for the management of pediatric neurosurgical disorders. Intraoperative imaging not only helps surgeons navigate through eloquent areas of the brain, but also ensures the maximal possible tumor resection or confirms adequate catheter placement prior to skin closure. The impact of this technology on long term survival is yet to be determined. PMID- 12570141 TI - Cranial surgery and navigation with a compact intraoperative MRI system. AB - Experience with a compact unit for intraoperative magnetic resonance imaging (iMRI) and integrated surgical navigation is presented. The system, commercially known as the PoleStar N-10 (Odin Medical Technologies, Yokne'am, Israel) includes a 0.12 Tesla permanent magnet that docks under a regular OR table. A passive infrared-based optical navigation system is included and the device can be controlled by the operating neurosurgeon and staff. We have operated on 93 patients using the PoleStar N-10. Diagnoses included glioma in 27, pituitary adenoma in 19, meningioma in 15, and others in 40. Lesions were removed from the skull base in 35 patients of whom 8 had posterior fossa masses. Surgery was affected by imaging in 51% of operations. Additional lesion, not otherwise apparent, was removed in 21; in 14 other patients unnecessary dissection was avoided when the new image confirmed that the surgical goals had been achieved. Additional time incurred with use of the PoleStar diminished with increasing experience. Testing of the navigation tool confirmed its accuracy to be comparable to other "frameless stereotactic" devices, on the order of 2 mm; likewise accuracy was increased by using T1 weighted imaging with thin slices. The introduction of new, faster imaging sequences and targeting tools has helped to make the PoleStar N-10 a routine tool for intracranial surgical navigation, with the added benefit of updated imaging during the procedure. PMID- 12570142 TI - Intraoperative ultrasound imaging: practical applicability as a real-time navigation system. AB - Experience with the use of Intaoperative Ultrasound (US) imaging as real time navigation system in neurosurgery is presented and discussed. Since 1987 we have performed US routinely in a wide variety of intracerebral and intramedullar pathologies. In this analysis we define useful intraoperative applications. Accurate definition of deep-seated lesions and their delineation from surrounding anatomical structures is possible with an US frequency of 5 MHz. Small subcortically located lesions can clearly be visualized with a high frequency probe. Differentiation between solid tumor, cyst and necrosis can be delineated. Identification of residual tumor is difficult. Dural sinuses and eventual invading tumor can be visualized by a 10 MHz probe. US guidance can be helpful for puncturing with a catheter, needle or endoscope. Postoperative percutaneous US imaging through a burr hole did not prove to be useful. The intraoperative use of US imaging is a reliable method for determining the size, shape and localization of lesions. It can be used as a practicable, cost effective and timesaving real time navigation system. PMID- 12570143 TI - Intraoperative ultrasound imaging: comparison of pathomorphological findings in US versus CT, MRI and intraoperative findings. AB - Since 1987, Ultrasound (US) is performed routinely as real time navigation system in our neurosurgical practice. In 374 cases with different pathologies the preoperative CT and MRI images were compared with the intraoperative US images and the operative findings. In all instances, the lesion could be localized and described in detail. US findings correlated with the findings an CT/MRI, concerning size and shape of lesions. US allowed the differentiation of more structural details within tissue compartments. The demarcation of gliomas was not as well defined in US as compared to CT/MRI, which correlated with the intraoperative situation. As for CT/MRI imaging, a correlation between US findings and histopathology of the lesion was not possible. In our opinion. intraoperative US imaging is an excellent tool for localization of cerebral and medullar lesions and for detailed description of their interior. This indicates a widespread applicability of this method in neurosurgery as an anatomical link between preoperative imaging and the reality of the operative field. PMID- 12570144 TI - CT-guided neurosurgery: preliminary experience. AB - BACKGROUND: With the possibility of CT systems becoming more handy and sophisticated, intraoperative CT was introduced in a few neurosurgical Centres with better results in lesion removal and surgical outcome. METHOD: At our Institution a mobile CT scanner was recently used for intraoperative evaluation (Philips Tomoscan M). For 27 tumour resections performed with a neuronavigation system, and 23 deep brain electrode positioning examinations, an intraoperative CT was employed. In addition the CT scanner was used in the recovery room for a postoperative control in 198 patients. FINDINGS: Our preliminary experience used for a real time evaluation of the treated patients, permitted to verify an incomplete removal in 23/27 cases. Evaluation of stereotactic electrode position in relation to the planned target was also possible and demonstrated a correct position in 21 cases. INTERPRETATION: Intraoperative CT scan is a useful system that permits to modify neuronavigation planning and is able to give information to the surgeon for better tumour removal, rule out possible hemorrhagic complications, and suitable deep brain electrode positioning. PMID- 12570145 TI - Intraoperative computerized tomography for improved accuracy of spinal navigation in pedicle screw placement of the thoracic spine. AB - We report on our experiences with the use of intraoperative CT imaging in surgery of the thoracic spine and on our results of pedicle screw insertion using spinal navigation and implantable fiducial markers. For our operations we used the Tomoscan M-EG and the EasyGuideSpine (Philips Medical Systems). During the operation the patient was positioned on the mobile CT table. Following dorsal preparation, small titanium screws were implanted in the vertebrae so as to serve as fiducial markers. Image data were obtained by performing a spiral CT scan. Ventilation was suspended for the duration of the CT scan. Screw insertion as well as vertebral biopsies were performed using spinal navigation. Intraoperative CT scans were obtained to confirm the position of the implants and to assess the amount of bony decompression as well as the realignment. Since 1998, 112 patients with various disorders of the thoracic spine have been operated on using the described technique. 365 screws were inserted in the area of T1 to T12. There were 23 (6.3%) misplacements of pedicle screws. In 42 cases (11.5%) we observed a minimal lateral perforation (<2 mm) of the pedicle wall. No neurological, cardiovascular, or pulmonary injury occurred. Intraoperative CT imaging influenced surgical decisions as well as the final result of surgery. Despite the use of intraoperative imaging and accurate spinal navigation, pedicle screw placement in the thoracic spine remains extremely challenging. PMID- 12570146 TI - Intraoperative imaging in a comprehensive neuronavigation environment for minimally invasive brain tumour surgery. AB - BACKGROUND: Development of an image-guided operation theatre offering multimodal information for mini-invasive neurosurgical brain tumour operations. METHODS: A multi-purpose resistive low-field MR scanner with on-off capability, was installed in a radio frequency-shielded operating room with in-room control panel and display. Intraoperative ultrasound imaging with Doppler mode as needed is used to provide check-up image data between intraoperative MR-imaging sessions. Cortical stimulation and registration are performed during awake craniotomies. The neuronavigation systems are customised arm-based and passive optical. The navigation systems show the positions of the ultrasound probe, cortical stimulation electrode, biopsy needles, endoscope and other instruments on the intraoperative MR-images. FINDINGS: Since 1999, 70 patients (mean age 47, range 3 88 years) have been operated with intraoperative MR-guidance (including 10 tumour biopsies, 56 resections). Twenty-one patients (mean age 46, range 16-67 years) underwent awake craniotomy and tumour resection secured with cortical stimulation and usually preoperative fMR-imaging. The present operating environment offered useful multimodal information for surgery of brain tumours in critical locations. Surgical mortality was 0%, morbidity included 3 (4.3%) infections and 2 (2.9%) permanent hemiparesis. Further removal of tumour was continued in 17 cases (57%) out of the 30 cases where intraoperative MR imaging was used for controlling completeness of the resection. PMID- 12570147 TI - Surgical navigation in the open MRI. AB - The introduction of MRI into neurosurgery has opened multiple avenues, but also introduced new challenges. The open-configuration intraoperative MRI installed at the Brigham and Women's Hospital in 1996 has been used for more than 500 open craniotomies and beyond 100 biopsies. Furthermore the versatile applicability, employing the same principles, is evident by its frequent use in other areas of the body. However, while intraoperative scanning in the SignaSP yielded unprecedented imaging during neurosurgical procedures their usage for navigation proved bulky and unhandy. To be fully integrated into the procedure, acquisition and display of intraoperative data have to be dynamic and primarily driven by the surgeon performing the procedure. To use the benefits of computer-assisted navigation systems together with immediate availability of intraoperative imaging we developed a software package. This "3D Slicer" has been used routinely for biopsies and open craniotomies. The system is stable and reliable. Pre- and intraoperative data can be visualized to plan and perform surgery, as well as to accommodate for intraoperative deformations, "brain shift", by providing online data acquisition. PMID- 12570149 TI - Costs and benefits of intraoperative MR-guided brain tumor resection. AB - We retrospectively compared the costs and benefits of brain tumor resection in the conventional operating room (cOR) with the interventional magnetic resonance (iMR) suite from 1993-1998. Comparisons were made for adults (diagnosis-related group (DRG) 001) and children (DRG 003) for length of stay (LOS), hospital charges and payments, hospital total direct and indirect costs, readmission rates, repeat resection (RR) interval, and net health outcome. Statistical analysis was with ANOVA, Dunnett's, and Bonferroni tests. For DRG 001, iMR LOS (3.7 days (d)) was 54.9% shorter than for cOR (8.2 d) for first resections (FR) (P < 0.001) and RR (6.0 vs. 8.7 d (31.0%), P < 0.05). IMR hospital charges were 12.2% lower ($4063) for FR and 4.1% lower ($922) for RR than for cOR. Total iMR hospital costs were 14.4% lower ($3415) than for cOR for FR and 3.3% lower ($723) than costs for RR. Cost-to-charge ratio (c/c) for FR was 69.6% (iMR) and 71.4% (cOR) and for RR 70.9% (iMR) and 71.1% (cOR). For DRG 003, iMR LOS (4.5 d) was shorter than for cOR (14.1 d, P < 0.001) for FR and for RR (8.0 vs. 13.3 d). IMR hospital charges were 43.8% lower than for cOR for FR (P < 0.05) and RR. The iMR costs were lower for FR (46.4%, P < 0.01) and RR (44.7%) than cOR. IMR c/c was 71.4% and 74.8% for cOR. For RR, the iMR c/c was 72.8% and 73.9% for cOR. No RR have followed iMR surgery. COR RR rate was 20% in adults and 30% in children. The mean time from iMR surgery was 11.3 months in adults and 18.0 in children. For the cOR, the mean time to RR was 9.3 months in adults and 13.3 in children. This data suggests that iMR surgery improves net health outcomes by reduced LOS, reduced RR, and reduced hospital charges and costs. PMID- 12570148 TI - The roles of functional MRI in MR-guided neurosurgery in a combined 1.5 Tesla MR operating room. AB - BACKGROUND AND PURPOSE: During MR-guided neurosurgical procedures performed in a combined 1.5 Tesla MR-operating room (MR-OR), we have successfully implemented and validated a functional MRI (fMRI) scheme for efficiently localizing eloquent functional areas and assessing their proximity to a lesion volume immediately prior to the craniotomy. METHODS: The fMRI examination consists of a dynamical blood oxygenation level dependent (BOLD) MR imaging technique and a task paradigm that is designed to activate the brain area of interest. The functional imaging technique was based on gradient-echo (GE) echo-planar imaging (EPI) (TR/TE = 2000 3000/40-50 msec). The motor task paradigm involves a periodic movement task, such as alternating between thumb and the other four fingers as a finger-tapping task, while the language involved a covert repeat of a series of words given as a task stimulus. While patient is performing the task, a dynamical fMRI was performed concurrently covering the volume of interest every 2 or 3 sec. Also, we have used a temporal series averaging (TSA) method for correcting the background drift in the raw fMRI signal, and developed a scheme for presenting fMRI results to neurosurgeons in an intuitive 3-dimensional volume-rendered display format. RESULTS: By using the fMRI scheme, we have successfully performed sixteen fMRI examinations immediately prior to neurosurgery in the combined MR-OR on the same surgical table to localize various eloquent functional areas of interests. TSA was successful in reducing the background drift in the fMRI time course data, and the 3-dimensional volume-rendered display was proven effective in presenting the resulting brain activations to neurosurgeons. More importantly, in three representative cases (one biopsy and two tumor resections) presented, the information provided by fMRI have indeed contributed significantly in making the optimal surgical decisions prior to craniotomy. CONCLUSIONS: Intra-operative fMRI can be an indispensable tool for determining the location of a neighboring eloquent functional area of concern in reference to a targeted lesion. Information provided by fMRI has helped in improving the outcome and clinician confidence of all surgeries performed. PMID- 12570150 TI - Intraoperative imaging in neurosurgery. MRI, CT, ultrasound. Postscript. PMID- 12570151 TI - Proteomics--advances, applications and the challenges that remain. PMID- 12570152 TI - Trends in mass spectrometry instrumentation for proteomics. AB - Mass spectrometry has become a primary tool for proteomics because of its capabilities for rapid and sensitive protein identification and quantitation. It is now possible to identify thousands of proteins from microgram sample quantities in a single day and to quantify relative protein abundances. However, the need for increased capabilities for proteome measurements is immense and is now driving both new strategies and instrument advances. These developments include those based on integration with multi-dimensional liquid separations and high accuracy mass measurements and promise more than order of magnitude improvements in sensitivity, dynamic range and throughput for proteomic analyses in the near future. PMID- 12570153 TI - Multidimensional peptide separations in proteomics. AB - Multidimensional peptide separation will play an increasingly important role in the drive to identify and quantitate the proteome. By increasing the peak and load capacity, multidimensional approaches increase the number and dynamic range of peptides that can be analyzed in a complex biological organism. Separation methods using different physical properties of peptides have been combined with varying degrees of success. The ultimate goal is a rapid separation strategy that can be coupled with analytical methods, such as mass spectrometry, to provide comprehensive monitoring of the changing concentration, interactions, and structures of proteins in the proteome. PMID- 12570154 TI - Protein chips: from concept to practice. AB - A series of exciting reports over the past two years has established the usefulness of protein chips and made important advances in preparing protein arrays. However, several technical challenges must still be addressed to make these tools available to the wider community of researchers. Here, we discusses these challenges and survey recent opportunities for creating quantitative assays, preparing and immobilizing large numbers of proteins, using detection methods to analyze the results of chip-based experiments, and using informatics tools to interpret these results. PMID- 12570155 TI - Affinity-capture reagents for protein arrays. AB - The simultaneous identification and quantitative measurement of the production levels of thousands of different proteins in a biological specimen remains an unachieved goal of modern proteomic research. Advances in the development of microarray-based platforms for highly parallel detection of proteins have therefore received a considerable impulse during the last few years. Here, we review the existing reagents for affinity capture of protein targets, as well as the techniques used for their immobilization on solid supports and methods for the detection of binding events, underlining the problems and the opportunities in this continuously evolving research field. PMID- 12570156 TI - Advances in quantitative proteomics using stable isotope tags. AB - A great deal of current biological and clinical research is directed at the interpretation of the information contained in the human genome sequence in terms of the structure, function and control of biological systems and processes. Proteomics, the systematic analysis of proteins, is becoming a critical component in this endeavor because proteomic measurements are carried out directly on proteins--the catalysts and effectors of essentially all biological functions. To detect changes in protein profiles that might provide important diagnostic or functional insights, proteomic analyses necessarily have to be quantitative. This article summarizes recent technological advances in quantitative proteomics. PMID- 12570157 TI - Proteomic analysis at the bedside: early detection of cancer. AB - Proteomic technologies promise to accelerate rapidly a new era in molecular medicine, especially in the detection and discovery of disease-related biomarkers. These technologies have no bigger impact than in the field of human cancer research. Beyond lifestyle-associated prevention strategies, early detection of cancer has the most profound impact on the ultimate course of the disease: the earlier the cancer is detected, the better the prognosis. Today, new proteomic technologies are being used to discover new diagnostic and prognostic biomarkers for the early detection and treatment of cancer that will have important implications at the bedside. PMID- 12570158 TI - Informatics and data management in proteomics. AB - Proteomics has become dominated by large amounts of experimental data and interpreted results. This experimental data cannot be effectively used without understanding the fundamental structure of its information content and representing that information in such a way that knowledge can be extracted from it. This review explores the structure of this information with regard to three fundamental issues: the extraction of relevant information from raw data, the scale of the projects involved and the statistical significance of protein identification results. PMID- 12570159 TI - Publishing large proteome datasets: scientific policy meets emerging technologies. AB - Currently, there are various approaches to proteomic analyses based on either 2D gel or HPLC separation platforms, generating data of different formats, structures and types. Identification of these separated proteins or peptide fragments is typically achieved by mass spectrometry (MS) measurements that use either accurate mass measurements or fragmentation (MS-MS) information. Integrating the information generated from these different platforms is essential if proteomics is to succeed. A further challenge lies in generating standards that can accept the hundreds-of-thousands of mass spectra produced per analysis based on threshold or probability measurements. Finally, peer review and electronic publication processes will be crucial to the dissemination and use of proteomic information. Merging the policy requirements of data-intensive research with information technology will enable scientists to gain real value from global proteomics information. PMID- 12570160 TI - Proteomics: drug target discovery on an industrial scale. AB - The discovery of targets that are sufficiently robust to yield marketable therapeutics is an enormous challenge. Through the years, several approaches have been used with varying degrees of success. These include target-independent screening of tumor-derived cell lines (disease-dependent), reductionist approaches to identifying crucial elements of disease-affected pathways, disease independent screening of homologs of previously drugged targets, disease dependent 'global' examination of gene transcript levels, and disease-dependent global examination of protein expression levels. These endeavors have been enabled by several major advancements in technology, most recently, the sequencing of the human genome. This review identifies the technical issues to be addressed for industrial-scale protein-based discovery in the identification of targets for therapeutic (or diagnostic) intervention. Such approaches aim to direct discovery in a way that increases the probability of robust target identification, and decreases the probability of failure owing to variable expression in this emerging field. PMID- 12570161 TI - Factor Va-factor Xa interactions: molecular sites involved in enzyme:cofactor assembly. AB - The generation of thrombin by the prothrombinase complex is a key event in coagulation. In this complex, the activated form of coagulation factor V (FVa) serves as an essential cofactor to factor Xa (FXa) in the activation of prothrombin to thrombin. The enzyme FXa is virtually ineffective in the absence of its cofactor. The assembly of FXa with its cofactor FVa on negatively charged phospholipid membranes enhances its catalytic efficiency by several orders of magnitude. The non-activated procofactor factor V (FV) circulates in plasma with a domain organization of A1-A2-B-A3-C1-C2 expressing little procoagulant activity. Upon activation through limited proteolysis by either thrombin or FXa, the B-domain dissociates from FVa. After activation, the procoagulant activity of FVa is greatly enhanced. This report provides insight into the interaction of FV and FXa and the molecular events important in enzyme:cofactor assembly of the FXa:FVa complex. Furthermore, light is shed on the molecular events associated with the activation process, i.e. the release of the B-domain and exposure of binding sites for FXa. The assembly of FVa and FXa was studied using a set of recombinant FV mutants. The interaction between FVa and FXa on phospholipid was investigated with a functional prothrombin activation assay as well as in a novel direct binding assay in the absence of prothrombin. We found that all three thrombin cleavages in FV contribute to increasing the FXa affinity and that the B domain in intact FV has an inhibitory effect on the FV-FXa interaction, which is important in prohibiting premature coagulation. PMID- 12570162 TI - Role of the N-terminal EGF module of coagulation factor IX in activation of factors IX and X. AB - Absence or reduced activity of coagulation factor IX (FIX) causes the severe bleeding disorder haemophilia B. FIX contains a Gla module, two epidermal growth factor-like (EGF) modules, and a serine protease region. I characterized a monoclonal antibody and found that it recognizes an epitope around residues 72 and 80 in the C-terminal part of EGF1 in human FIX. The antibody exhibited 10 fold greater affinity for activated FIX (FIXa) than for the zymogen FIX, indicating the existence of intra-molecular communication between the serine protease region and EGF1. Binding of the antibody did not affect the amidolytic activity of FIXa, hence I could use the antibody during activation of FIX to show that the C-terminal part of EGF1 is of importance for the interaction with FXIa but not with FVIIa/TF. Considering activation of FX, it is a matter of debate whether EGF1 or FIXa interacts directly with FVIIIa. I activated FX in the presence and absence of the antibody and/or FVIIIa. The addition of antibody caused only a minor decrease in k(cat,app), and the major increase in k(cat,app) caused by the addition of FVIIIa occurred even in the presence of the antibody. This implies that EGF1 of FIXa is not directly involved in interaction with FVIIIa in the Xase complex. A model of the FIXa-FVIIIa complex, based on my findings and results from the literature, was constructed and indicated that EGF1 of FIXa does not interact directly with FVIIIa. PMID- 12570163 TI - Studies on the interaction between vitamin K-dependent protein S and complement regulator C4b-binding protein: localization of binding sites and identification of a possible function of the complex. AB - Complement is a cascade-like system that is part of the innate immune defence. It is an explosive system, potentially harmful also for the host cells, and needs to be strictly regulated. An important down-regulator of complement is C4b-binding protein (C4BP). C4BP contains two different types of subunits, seven identical alpha-chains and one unique beta-chain. The alpha-chains bind to C4b, C4BP's target in the complement system. The beta-chain binds to vitamin K-dependent protein S. Approximately 70% of all protein S in plasma circulates in a high affinity complex with C4BP. Free protein S, the remaining 30%, functions as an important cofactor in the anticoagulant system. The reason for the complex formation between C4BP and protein S has remained an intriguing enigma. Protein S has a very high affinity to negatively charged phospholipids for protein S. One area where such phospholipids are present is the surface of the apoptotic cell, where the exposure of phosphatidylserine is an early event. Physiological apoptosis is characterized by a lack of inflammatory response in surrounding tissues, indicating that cells are rapidly cleared before leaking cytoplasmic components into the extracellular space. A number of studies demonstrate that early complement proteins are important for the removal of apoptotic cells, but that subsequent assembly of later complement components and anaphylatoxin release must be prohibited in order not to provoke an inflammatory response. We demonstrate that protein S localizes C4BP to the surface of apoptotic cells via binding to the exposed phosphatidylserine. The C4BP attached to the apoptotic cell through protein S was still able to bind C4b, suggesting that C4BP retains its physiological function also when localized to the apoptotic cell surface. In addition, we have also pinpointed a hydrophobic binding site for protein S on C4BP. The binding studies between C4BP and protein S were performed on recombinant proteins where mutations had been introduced. Mutations were chosen based on a 3D-homology model of the C4BP beta-chain. PMID- 12570164 TI - Soluble CD163: a marker molecule for monocyte/macrophage activity in disease. AB - By immunoprecipitation we have identified a soluble plasma form of CD163 (sCD163), the IL-6 inducible macrophage-receptor for clearing haptoglobin haemoglobin complexes. A sandwich ELISA for measuring sCD163 was established and used to determine the sCD163 levels in normal subjects and patients with inflammatory and myeloproliferative diseases. In normal subjects, the concentration of sCD163 was high (median 1.9 mg/l) with low intra-individual variation. Highly increased levels were seen in patients with sepsis, myeloid leukaemia and in patients with Gaucher disease characterized by accumulation of tissue macrophages. Although the physiological role of sCD163 remains unknown, our present data suggest that sCD163 might prove to be a valuable marker molecule in infectious and myeloproliferative diseases. PMID- 12570165 TI - Lipoprotein production by the heart: a novel pathway of triglyceride export from cardiomyocytes. AB - The current knowledge on lipoprotein secretion from the heart is examined in this article. The ability of cells to secrete apolipoprotein B (apo-B) containing lipoproteins depends on expression of the apo-B and microsomal triglyceride transfer protein (MTP) genes. Initially, it was shown that both genes are expressed in cardiac myocytes of mice and humans. Electron microscopy of human cardiac myocytes revealed lipoproteins in the secretory pathway and metabolic labelling studies demonstrated the secretion of LDL-like lipoproteins from minced heart biopsies. To examine the role of lipoprotein formation in the heart, we tested whether overexpression of a human apo-B transgene in the heart affects cardiac triglyceride accumulation. In wild-type mice, diabetes conferred an increase in heart triglycerides. In apo-B transgenic mice, diabetes did not affect heart triglycerides. Also, apo-B overexpression prevents fasting-induced heart triglyceride accumulation, whereas inhibition of MTP expression increases heart triglycerides in mice. In hypoxic human hearts, MTP mRNA expression was negatively associated with triglyceride contents. These findings suggest that lipoprotein formation rates affect cardiac triglyceride stores. The MTP mRNA levels are approximately 2-fold higher in hypoxic compared with normoxic human myocardium and in diabetic compared with non-diabetic mouse hearts. In both hypoxia and diabetes, the delivery of triglycerides to the heart exceeds their utilization for beta-oxidation. Thus, endogenous lipoprotein secretion rates might be upregulated to remove surplus fat from the heart. Diabetes negatively affected indexes of systolic and diastolic function in wild-type mice. However, the diabetogenic effects on the heart were absent or much less pronounced in apo B transgenic mice. This suggests that accelerated lipoprotein formation by the heart attenuates development of diabetic cardiomyopathy in mice. In conclusion, current evidence suggests that lipoprotein secretion from the heart plays an integrated role in cardiac lipid homeostasis and that it can affect the biomechanical function of the heart. PMID- 12570166 TI - Aggressive angiomyxoma of the pelvis: case report and review. AB - OBJECTIVE: To report a case of recurrent aggressive angiomyxoma managed by a team composed of a radiologist, general surgeon, and reproductive endocrinologist, with a literature review which focuses on histologic differences between various types of myxomas. STUDY DESIGN: Case report and literature review. RESULTS AND CONCLUSIONS: The proband patient has an apparent cure, but this particular type of myxoma shows a proclivity for recurrence, sometimes years after resection. Therefore, long-term follow-up with MRI or CT scans is necessary. Preoperative management with vessel embolization and creation of gonadal suppression facilitates the surgical approach, which usually can be via a perineal, extraperitoneal route. PMID- 12570167 TI - Triplet pregnancies and their aftermaths. Part I: Basic considerations. AB - Until recently, triplets were an infrequent occurrence. With the development of ovulation-inducing medications and then assisted reproductive technology (ART), the triplet birth rate in maternal age ranges over 24 years has multiplied three- to fiftyfold. The first review of the clinical aspects of triplet pregnancies was published in 1988. Since then, more data reported on such births have revealed a consistent triad of preterm delivery, low birth weight, and very low birth weight, plus increased maternal risks. This article details comparative risks to mothers and children using population-based data comparing differential outcomes in singletons, twins, and triplets; an extensive review of recent triplet publications is provided in the references. PMID- 12570168 TI - Positive bacterial culture of semen from infertile men with asymptomatic leukocytospermia. AB - OBJECTIVE: To examine results of semen culture in a population of infertile men with asymptomatic leukocytospermia, and to determine the correlation between culture results and sperm characteristics in these patients. METHODS: Semen samples were collected from a group of infertile men (n = 80) after 2-3 days of sexual abstinence. Standard semen analysis was performed according to World Health Organization (WHO) guidelines. Seminal leukocyte concentrations were determined by a myeloperoxidase staining technique. Culture of semen was performed on enriched and specific culture media. RESULTS: Based on the results of semen culture, the samples were classified into three groups: group 1 (enteric gram-negative bacilli [Enterobacteriaceae], n = 13), group 2 (bacteria other than Enterobacteriaceae, n = 15), and group 3 (negative cultures, n = 52). No correlation was found between positive semen culture and sperm characteristics (concentration, motility, and morphology). However, seminal leukocyte concentrations were negatively correlated with percentages of motile sperm and normal sperm morphology. CONCLUSION: Our data indicate that semen samples from infertile men with asymptomatic leukocytospermia may have bacterial contamination, evidenced by positive bacterial cultures. Lack of correlation between positive semen cultures and sperm characteristics may be indicative of early or mild (subclinical) infection. PMID- 12570169 TI - A prospective, double-blind, randomized, placebo-controlled clinical trial of bromocriptine in clomiphene-resistant patients with polycystic ovary syndrome and normal prolactin level. AB - OBJECTIVE: To evaluate the effects of administration of bromocriptine combined with clomiphene citrate (CC) in CC-resistant patients with polycystic ovary syndrome (PCOS) and normal prolactin (PRL) level. DESIGN: Prospective double blind, placebo-controlled, randomized. SETTING: Referral university hospitals. PATIENTS: One hundred women with PCOS and normal PRL who failed to ovulate with a routine protocol of CC. INTERVENTIONS: Treatment group received 150 mg of CC from day 5 to 9 and 7.5 mg bromocriptine continuously, with hCG 10,000 units on day 16 or 17. Control group received the same protocol of CC combined with placebo. MAIN OUTCOME MEASURES: Follicular development, hormonal changes, ovulation rate, pregnancy rate. RESULTS: Follicular development (follicular size greater than 15 mm) was observed in 12 (25.5%) and 8 (15.1%) women in the treatment and placebo group, respectively (p = 0.29). The serum prolactin level was within normal limits in all patients before treatment. After 3 and 6 months of treatment with bromocriptine, there was a significant decrease in serum level of prolactin (p = 0.000001). No significant differences were seen in ovulation, pregnancy rate, or serum levels of FSH, LH, DHEAS, and progesterone between treatment and placebo groups after treatment. CONCLUSIONS: The only significant effect of long-term bromocriptine therapy in CC-resistant women with PCOS was to lower the serum PRL concentration. It is also concluded that 10%-15% of patients with PCOS experienced occasional ovulatory cycles and pregnancy whether or not they were on treatment. PMID- 12570170 TI - Peer education for HIV prevention in the Socialist Republic of Vietnam: a national assessment. AB - In 2000, the government of Vietnam conducted the first assessment of its national peer education program for HIV prevention. Twenty (32%) of Vietnam's 61 provinces and urban areas had functioning peer education programs, and program coordinators of all 20 were interviewed regarding their programs. In addition, on-site reviews were done for 10 of the 20 programs, including interviews of peer educators and high-risk persons in each program. The assessment found that a total of 500 peer educators were functioning either independently or as part of one of 79 teams. In the 20 provinces, the peer educators made an estimated 7,000 total contacts per month with high risk persons, but many persons were likely contacted repeatedly. Despite this, coverage was limited: some provinces with high numbers of persons reported with HIV/AIDS had few peer educators. Although most provinces targeted IDU and many targeted CSW, few provinces targeted sex partners of IDU or CSW. The definition of peer education and composition of teams varied substantially by province; only one province included persons living with HIV/AIDS as peer educators. The services provided by peer educators were primarily distributional: delivering information either through word of mouth, pamphlets, or brochures, providing condoms, and sometimes providing clean syringes and needles. Skills building or goal setting interventions aimed at HIV risk reduction were rarely provided. Most provinces had concerns about ongoing funding and sustainability of the programs. Based on the assessment, specific recommendations were provided for strengthening and expanding Vietnam's peer education programs. PMID- 12570171 TI - Human cargo: health conditions of Chinese migrants interdicted offshore by U.S. authorities. AB - During the eight month period between April and December 1999, the United States Coast Guard intercepted seven boats carrying migrants from the People's Republic of China destined for the United States. These migrants were processed by the United States Immigration and Naturalization Service in three locations: Tinian Island, Midway Island, and Guatemala. Emergency Medical Response Teams from the United States Public Health Service, Division of Immigration Health Services, were deployed to conduct initial health screenings of the 913 migrants on board these ships and provide on-going health care until the individuals were repatriated or relocated. The distributions of demographic characteristics of the population and the health conditions observed are presented. Differences in health conditions observed by temporary detention location, sex, and age group were assessed. The majority of migrants were males younger than age 30. Few serious illnesses were observed. The most prevalent conditions included skin rashes, fungal rashes, upper respiratory infections, abdominal discomfort, scabies, abrasions, skin lesions, headache, pain and/or injuries, dental problems, and ear problems. For many health conditions, statistically significant differences were observed by location. For nearly all conditions for which differences were observed by sex, these differences were accounted for by a greater proportion of females presenting with the condition. PMID- 12570172 TI - Knowledge of cervical cancer risk factors among Chinese immigrants in Seattle. AB - Chinese American immigrants are a growing part of the United States population. Cervical cancer is a significant cause of morbidity and mortality among Chinese Americans. Pap smear testing is less common in Chinese American immigrants than in the general population. During 1999, we conducted a community-based survey of Chinese American women living in Seattle. We assessed knowledge of cervical cancer risk factors and history of Pap smear testing along with socioeconomic and acculturation characteristics. The overall estimated response rate was 64%, and the cooperation rate was 72%. Our study sample included 472 women. Most cervical cancer risk factors were recognized by less than half of our participants. Factors independently associated with knowledge of cervical cancer risk factors included marital status, employment, and education. Respondents with the highest knowledge had greater odds of ever receiving a Pap smear, compared to those respondents with the lowest knowledge (OR 2.5; 95% CI: 1.1,5.8). Our findings suggest a need for increased recognition of cervical cancer risk factors among Chinese American immigrants. Culturally and linguistically appropriate educational interventions for cervical cancer risk factors should be developed, implemented and evaluated. PMID- 12570173 TI - Evaluation of access, a primary care program for indigent patients: inpatient and emergency room utilization. AB - We analyzed the impact of a program that provides indigent patients with free primary care on inpatient admissions, emergency room (ER) visits, and resulting charges in 91 patients before and after admittance into the program. There was a decrease in ER visits after enrolling in the program (1.89 versus 0.83 visits per year; p < 0.0001). This difference translated into mean ER charges of $1,174 vs. $717 (p = 0.0007), and a decrease in charges of $41,587 per year. The charges for the program (outpatient visits and laboratory) were $23,141. Entry into the program had no effect on inpatient admissions, which averaged 0.07 admissions per year both before and after admission to the program. Indigent patients enrolled in a complimentary primary care program had significantly decreased per-year ER utilization rates and charges. The program had no effect on inpatient admissions. By conservative estimate, the program decreased ER charges by approximately $18,000 per year secondary to decreased ER utilization. PMID- 12570174 TI - Sale of alcohol in pharmacies: results and implications of an empirical study. AB - Alcohol misuse continues to be a major concern in the health care community. Within this community, pharmacists are ideally positioned to play a major role in preventive efforts in dealing with alcohol misuse. The objective of the study was to follow-up on important findings from a 1996 statewide survey of Indiana pharmacists regarding their opinions and practices related to the sale of alcohol in pharmacies. Of the 1,280 registered Indiana pharmacies, one-half were chosen using a systematic random selection process. From the 640 pharmacies contacted, 476 pharmacists responded completely to the questionnaire, providing a 74.4% response rate. Findings reveal that independent pharmacies had significantly reduced their sale of alcohol since 1996 while there was no significant change among retail chain pharmacies. Pharmacies that generated a greater share of their business from prescription drugs were less likely to sell alcohol products. Compared to five years ago, significantly more pharmacists in 2001 believed that pharmacies should not sell alcohol. In addition, more than half of pharmacists who worked in pharmacies that sold alcohol felt it differed from their professional values. Twenty-three percent of pharmacists reported inquiring about their patients' alcohol habits, a significant increase over 1996. More pharmacists thought alcohol was an important contributing factor to morbidity in 2001 compared with 1996. The data also revealed that pharmacists were more likely to ask patients about their alcohol habits if they considered alcohol use to be a more important cause of morbidity. PMID- 12570175 TI - Evidence-based practice. PMID- 12570176 TI - Evidence-based practice. PMID- 12570177 TI - Vaginal douching. AB - OBJECTIVE: To review current literature on vaginal douching. DATA SOURCES: MEDLINE, CINAHL, and Cochrane databases from 1997 to 2001, using keywords douche or douching; 2001 Web sites of the Centers for Disease Control and Prevention; and Internet search engines for information about current retail sales of douches. STUDY SELECTION: MEDUNE included 67 records, CINAHL 18, and Cochrane 2. Abstracts of articles in English were reviewed, and those pertaining to vaginal douching practices were included. MEDLINE had 44 pertinent articles, CINAHL 11, and Cochrane 1. References from these articles were reviewed and included when appropriate. DATA EXTRACTION: Articles were reviewed and summarized. DATA SYNTHESIS: Vaginal douching is a common practice for women in the United States. Douching is associated with adverse reproductive and gynecologic outcomes including bacterial vaginosis, preterm birth, low-birth-weight infants, pelvic inflammatory disease, chlamydial infection, tubal pregnancy, higher rates of HIV transmission, and cervical cancer. Cultural beliefs and educational factors strongly influence douching practices. CONCLUSIONS: Nursing assessment of women should include information on vaginal douching practices and beliefs. Nurses should use culturally appropriate educational strategies to discourage women of all ages from using vaginal douches as part of routine feminine hygiene because of the associated risks. Further research is needed on factors that influence women's beliefs and douching practices. PMID- 12570178 TI - The graying of the obstetric population: implications for the older mother. AB - OBJECTIVE: To review the literature on the relationship between advanced maternal age and pregnancy outcomes and to discuss social trends in childbearing. DATA SOURCES: Computerized searches on CINAHL, MEDLINE, National Institutes of Health, and published birth statistics. Keywords used in this search were the following: advanced maternal age, pregnancy outcomes for older primiparae and older mothers, and pregnancy risks. STUDY SELECTION: Relevant articles published between 1978 and 2001 were reviewed. DATA EXTRACTION: Data were extracted and organized under the following headings: childbearing trends in developed countries, maternal morbidity for older mothers, preeclampsia, preterm delivery, operative and assisted delivery, and infant mortality and morbidity. DATA SYNTHESIS: The existing literature is concentrated on the medical aspects of the risks associated with childbearing at an advanced maternal age. Few systematic investigations have been undertaken of healthy older mothers, an increasingly common obstetric population. CONCLUSIONS: Lifestyle influences, such as weight, general health, and smoking habits in older mothers, have not been addressed. Little is known about the actual risks of pregnancy complications for healthy older mothers. No clear guidelines exist to direct the obstetric and nursing care of the healthy older mother. PMID- 12570179 TI - Coping with grief after involuntary pregnancy loss: perspectives of African American women. AB - OBJECTIVE: To present the coping strategies used by African American women following their miscarriages, ectopic pregnancies, fetal deaths, and still births, which the authors have termed involuntary pregnancy losses or IPLs. DESIGN: Semistructured audiotaped interviews; grounded theory methods used to collect and analyze the data. SETTING: Urban community-based sites in the Western United States. PARTICIPANTS: 20 African American adult women who reported a history of involuntary pregnancy loss within 3 years of interview. RESULTS: In this study, the women's responses to their IPL were grouped into four areas. They coped with personal reactions, reactions of others, memories of the baby, and subsequent pregnancies. CONCLUSION: The women in this study used inner resources to develop self-help strategies to cope with reactions following IPL. Nurses are challenged to harness the influence of family, friends, religion, and cultural traditions to assist women in processing the cognitive, emotional, and social traumas associated with IPL. Educating women to recognize grief responses after IPL and to manage these responses effectively may prevent adverse outcomes to their physical and mental health. A culturally sensitive framework of clinical assessment and intervention for African American women experiencing IPL has been developed. PMID- 12570180 TI - Breastfeeding knowledge, breastfeeding confidence, and infant feeding plans: effects on actual feeding practices. AB - OBJECTIVE: To explore relationships among breastfeeding knowledge, breastfeeding confidence, and infant feeding plans and their effects on feeding practices in first-time breastfeeding mothers. DESIGN: Prospective descriptive design. SETTING: Telephone interviews were conducted prenatally and at 6 weeks, 3 months, and 6 months postpartum. PARTICIPANTS: Seventy-four of 83 first-time mothers with prenatal intentions to breastfeed completed all study requirements. The majority were White (95%), between the ages of 21 and 30 years (73%), with a post-high school education (85%), and household incomes of more than 200% of the federal poverty guideline (88%). MAIN OUTCOME MEASURES: Breastfeeding knowledge, breastfeeding confidence, planned infant feeding method, planned breastfeeding duration, weeks of daily human milk substitute feeding, breastfeeding duration, achievement of breastfeeding goals. RESULTS: Breastfeeding knowledge was strongly correlated with breastfeeding confidence (r = .262; p = .025) and actual lactation duration (r = .455; p = .0001). Compared with women planning to exclusively breastfeed their infants, those planning to combination feed planned shorter breastfeeding duration (p = .022), reported shorter actual duration (p = .004), and were less likely to meet their breastfeeding goal (p = .034). The variables maternal education, breastfeeding knowledge, and weeks of daily human milk substitute feeding were used to develop a prediction equation that correctly categorized 93% of participants who met their breastfeeding goal and 90% of those who did not. CONCLUSIONS: Expectations and the actual breastfeeding experience differed among women planning to combination feed and those planning to exclusively breastfeed. Whether a cause or consequence, daily human milk substitute feeding was associated with negative breastfeeding outcomes. PMID- 12570181 TI - Hypoglycemia and the full-term newborn: how well does birth weight for gestational age predict risk? AB - OBJECTIVE: To determine whether anthropometric characteristics could be used to accurately predict risk of hypoglycemia in full-term newborns during the early post-birth period. DESIGN: Descriptive, utilizing newborn anthropometric measurements singly and in combination to determine risk of neonatal hypoglycemia. The following measurements were obtained twice for each newborn: weight, head circumference, chest circumference, abdominal circumference, mid-arm circumference, thigh circumference, and length. The investigator was blind to all measurements except weight. SETTING: Mothers' rooms or the newborn nursery in a community hospital. INTERVENTIONS: All measurements were obtained twice, and a physical examination was completed on each newborn by the principal investigator. These newborns were classified as large-, average-, and small-for-gestational age, using a tool typically used in many newborn nurseries. SAMPLE: One hundred fifty-seven full-term newborns (94 White and 63 African American). MAIN OUTCOME MEASURES: The differences in anthropometric measurements by race and gender were calculated using two-way analysis of variance. The risk of hypoglycemia was calculated using logistic regression modeling. RESULTS: There were significant differences in measurements by race and by gender. Additionally, there was a subset of newborns classified as average for gestational age who had an increased risk of hypoglycemia (OR = 4.17, 95% CI = 1.33-13.08). Newborns with a mid-arm circumference/head circumference ratio that varied from .26 to .29 have an odds ratio of 6.10 (95% CI = 1.89-19.66) for risk of hypoglycemia. Plotting a newborn's birth weight on a published fetal growth curve clearly did not accurately predict his or her risk of hypoglycemia. CONCLUSIONS: These findings indicate that extremes in newborn birth weight are not always correctly defined, may vary by race and ethnic group, and may not be the best method for determining under- or overnourishment and risk of neonatal hypoglycemia. PMID- 12570182 TI - Keeping safe: teenagers' strategies for dealing with perinatal violence. AB - OBJECTIVE: To identify strategies adolescents use to cope with and keep safe from perinatal violence. DESIGN: Exploratory descriptive design incorporating qualitative content analysis. A semistructured interview guide was used to inquire about coping strategies. SAMPLE: 20 nonpregnant teenagers between the ages of 18 and 20 years who experienced violence before, during, or after a prior pregnancy, regardless of outcome, were recruited for this study from community advertising and two outpatient gynecologic clinics. RESULTS: The teenagers described experiences of rape, stalking, and physical and sexual violence. They identified perpetrators as family members, nonintimate social acquaintances, and male and female intimate partners. Five themes were identified that described the teenagers' experiences with "Keeping Safe," including Losing Ground: Moving Backward; Minimizing Damage: Decreasing the Impact and the Severity of Violence; Keeping Away: Drawing Inward and Moving On; Getting Help: Reaching Out to Others; and Taking Charge: Being Proactive. CONCLUSIONS: The teenagers were actively engaged in developing creative strategies to minimize stress and the severity of violence in their lives. They also incorporated maladaptive behaviors that further increased their jeopardy. Asking about the ways that teenagers deal with violence might assist health care workers to proactively minimize damage from ineffective coping mechanisms. PMID- 12570183 TI - Stages of change-based nursing interventions for victims of interpersonal violence. AB - Recently, the health care system has become recognized as an important site for domestic violence programs. However, most of these programs focus on screening for violence. There is a need for conceptually based intervention strategies that provide help for victims once they are identified. The Transtheoretical Model of Change is useful in suggesting interventions that are consistent with the ways victims perceive their situations and take actions to end violence in their lives. PMID- 12570184 TI - Intrapartum nursing: integrating Rubin's framework with social support theory. AB - Numerous studies and articles on labor support have focused on the potential for improved labor and birth outcomes from supportive care during labor. Despite increased attention to labor support research, surprisingly little has been written about the theoretical underpinnings for intrapartum nursing care. This article explores Reva Rubin's framework and social support theory as a foundation for intrapartum nursing care. The most common features of social support provided the structure into which Rubin's descriptions of nursing care during labor and birth could be evaluated. Social support theory fit remarkably well, both with Rubin's views of the role of the perinatal nurse and with Rubin's observations of mothers' needs and feelings during childbirth. Combining these theoretical frameworks provides a perspective of intrapartum nursing practice that has not been previously considered in the published literature. PMID- 12570186 TI - HIV update. PMID- 12570187 TI - Gynecologic care for women with HIV infection. AB - The purpose of this article is to review the gynecologic conditions encountered among women with HIV and to outline elements of gynecologic care for these women. The most prevalent problems are vulvo-vaginal candidiasis and cervical dysplasia; however, other sexually transmitted diseases, pelvic inflammatory disease, genital ulcer disease, and menstrual abnormalities are also seen. Drug interactions may limit the effectiveness or increase the toxicities associated with hormonal contraception. Intrauterine devices are contraindicated. PMID- 12570188 TI - Psychosocial correlates of HIV risk behavior in adolescent girls. AB - OBJECTIVE: To present the preintervention data collected for a pilot HIV prevention randomized controlled trial specifically developed for single, sexually active adolescent girls. DESIGN: Comparative, descriptive design using confidential self-administered questionnaires. SETTING: An urban family planning clinic that provided gynecologic services to adolescents. PARTICIPANTS: One hundred twenty-nine single, sexually active adolescent girls 15 to 19 years of age (44% minority, 34% economically disadvantaged). MAIN OUTCOME MEASURES: In addition to demographics and risk behaviors, the following were assessed: HIV related information (i.e., knowledge) and motivation to reduce risk (i.e., perceived vulnerability, readiness to change HIV risk behaviors, behavioral intentions, pros and cons of condom use, and confidence in condom use). RESULTS: Assessments revealed limited HIV-related knowledge, ambivalence regarding risk reduction, and considerable risk behaviors. Compared with girls at lower risk for HIV infection (i.e., consistent condom users), those at higher risk (i.e., inconsistent or noncondom-users) were more likely to be White and older and to have better HIV-related knowledge but less motivation to reduce risk (i.e., behavioral intentions to use condoms, condom-use confidence) than girls at lower risk. CONCLUSION: These data document (a) the need for HIV prevention interventions targeted to all sexually active adolescent girls and (b) the importance of including a motivational component in the intervention. PMID- 12570189 TI - The evolving art of caring for pregnant women with HIV infection. AB - Care for the pregnant woman who has human immunodeficiency virus (HIV) is highly complex and constantly evolving. The purpose of this article is to describe the pertinent issues regarding women and HIV during pregnancy, including epidemiology of mother-to-child transmission, the effect of the disease on pregnancy, care issues in the perinatal period, and the issues in women's lives that place them at risk for HIV infection. Antiretroviral therapy offers significant reduction in the rate of mother-to-child transmission, and this is presently the cornerstone of therapy for the pregnant woman with HIV. Careful intrapartum management may also reduce the risk of transmission. Clinical studies of treatment modalities continue to offer new hope to prevent transmission of the virus to the fetus. PMID- 12570190 TI - HIV exposure: neonatal considerations. AB - The presence of human immunodeficiency virus (HIV) in pregnant women puts infants at risk for exposure through placental infection and contact with contaminated maternal blood and genital secretions. Efforts to combat this inevitably fatal disease continue to focus on preventing transmission of the virus from a mother who has HIV to her newborn during the prenatal, intrapartum, and postnatal periods. Prophylaxis against transmission and vigilant assessment for indicators of infection are hallmarks of appropriate health care for infants exposed to HIV. PMID- 12570191 TI - HIV and infant feeding: issues in developed and developing countries. AB - Women in developed countries who are human immunodeficiency virus (HIV)-positive generally feed their infants formula to avoid virus transmission via breastfeeding. However, for most of the world's women who are HIV positive, the choice of infant feeding method is not so clear. Poor socioeconomic and living conditions place infants on breast milk substitutes at higher risk of non-HIV infectious diseases as compared with breastfed infants. Mothers in these settings who are HIV positive must weigh the risks and benefits of breastfeeding to choose the best infant feeding option. PMID- 12570192 TI - Searching for graves using geophysical technology: field tests with ground penetrating radar, magnetometry, and electrical resistivity. AB - Field experiments were conducted using three types of geophysical equipment in a variety of situations. The goal of the study was to ascertain the relative utility of this technology for non-geophysical expert forensic professionals searching for buried human remains. The study concludes that the equipment should be used with caution after a critical evaluation of specific field conditions, and more refinement of technical methods and skills should be developed. PMID- 12570193 TI - Measurement of eyeball protrusion and its application in facial reconstruction. AB - The estimation of eyeball protrusion from the bony orbit has traditionally relied upon Wolff's theory (1976) that the cornea will be tangental to a line taken from the superior to the inferior orbital margin. This study tested this theory by taking measurements from MRI cranial images of 78 eyes of white adult subjects. Orbital depth had a significant negative linear correlation with eyeball protrusion, and protrusion of the eyeball was determined by the following standard: eyeball protrusion = 18.3 - (0.4 x orbit depth). Current eyeball position determination was shown to be as much as 3.9 mm too deep in the socket, and a practical application of this result to facial reconstruction is discussed. PMID- 12570194 TI - Testing the validity of metacarpal use in sex assessment of human skeletal remains. AB - To assess the potential of employing metacarpals in assessing sex of human skeletal remains, previous investigators have generated regression equations (Scheuer & Elkington, 1995) and linear discriminant functions (Falsetti, 1995; Stojanowski, 1999) based upon measurements from metacarpals. Results have varied in overall accuracy and which metacarpal produces the greatest accuracy. Using a contemporary sample, this study seeks to evaluate the validity of using metacarpals to assign sex by testing methodologies of previous studies. Measurements defined by previous authors were repeated on metacarpals from 23 adult cadavers and data were subjected to regression equations and linear discriminant analysis according to previous methodologies. Accuracy in sex determination from methods of Scheuer & Elkington (1993) and Falsetti (1995) were lower than originally reported while accuracy from methods of Stojanowski (1999) were higher than previously reported. These results suggest that the use of metacarpals in sex determination may be limited and should be applied cautiously. PMID- 12570195 TI - The feasibility of external blind DNA proficiency testing. I. Background and findings. AB - We describe the origins, purposes, and findings of a national study to determine whether a large-scale program of blind proficiency testing in U.S. DNA laboratories is feasible and/or practical. Proficiency testing in clinical laboratories is reviewed, particularly as mandated by the Clinical Laboratory Improvement Acts and its role in the regulation of those laboratories. Proficiency testing in forensic urine drug testing labs is also briefly reviewed. Studies involving comparisons between open and blind proficiency testing are discussed. The clinical laboratory proficiency testing and regulation landscape provides the background for the DNA Act of 1994, and the congressional mandate to investigate blind proficiency testing in forensic DNA laboratories. Four models of blind proficiency testing are defined and discussed, along with the advantages and disadvantages of each and estimates of the costs of a large-scale program. The purposes of proficiency testing in a quality-assurance context are likewise discussed and related to the models and the arguments generally proffered for and against blind vs. open proficiency testing. PMID- 12570196 TI - The feasibility of external blind DNA proficiency testing. II. Experience with actual blind tests. AB - The background and goals of a national study to determine the feasibility of blind proficiency testing in U.S. forensic DNA laboratories are discussed. Part of the project involved designing and executing a series of fifteen blind proficiency tests. Execution included biological specimen donor recruitment and case evidence manufacturing. Simulated cases were submitted to DNA laboratories by law enforcement agencies and in some cases by other forensic-science laboratories. Replicate-manufactured evidence was submitted to reference laboratories to simulate the workings of a larger-scale program. Ten tests were straightforward, and essentially tested analytical ability. Five tests involved selecting on the basis of case facts appropriate bloodstains for typing from a bloodstain pattern. We describe in detail our experience in designing and conducting these blind proficiency test trials, and relate those experiences to the overall issue of blind proficiency testing as a quality-assurance tool in forensic DNA laboratories. In this feasibility test series, one blind test was detected by a laboratory, a second one was shown to the lab by law enforcement, and a third was never completed because of lapses in communication. Turnaround times were relatively fast in the independent/commercial labs and relatively slow in the larger public laboratories. Two cross-state case-to-case CODIS "hits" were "planted" among the first series of ten blind tests. One pair was detected. One member of the second pair went to a lab that was not CODIS-ready. PMID- 12570197 TI - GC-MS of ignitable liquids using solvent-desorbed SPME for automated analysis. AB - Solid-phase microextraction (SPME) is well documented with respect to its convenience and applicability to sampling volatiles. Nonetheless, fire debris analysts have yet to widely adopt SPME as a viable extraction technique, although several fire debris studies have demonstrated the utility of SPME coupled with gas chromatography-mass spectrometry (GC-MS) to identify ignitable liquids. This work considers the expansion of SPME sampling from the customary thermal desorption mode to solvent-based analyte desorption for the analysis of ignitable residues. SPME extraction fibers are desorbed in 30 microL of nonaqueous solvent to yield a solution amenable to conventional GC-MS analysis with standard autosampler apparatus. This approach retains the advantages of convenience and sampling time associated with thermal desorption while simultaneously improving the flexibility and throughput of the method. Based on sampling results for three ignitable liquids (gasoline, kerosene, anddiesel fuel) in direct comparisons with the widely used activated charcoal strip (ACS) method this methodology appears to be a viable alternative to the routinely used ACS method. PMID- 12570198 TI - How the probability of a false positive affects the value of DNA evidence. AB - Errors in sample handling or test interpretation may cause false positives in forensic DNA testing. This article uses a Bayesian model to show how the potential for a false positive affects the evidentiary value of DNA evidence and the sufficiency of DNA evidence to meet traditional legal standards for conviction. The Bayesian analysis is contrasted with the "false positive fallacy," an intuitively appealing but erroneous alternative interpretation. The findings show the importance of having accurate information about both the random match probability and the false positive probability when evaluating DNA evidence. It is argued that ignoring or underestimating the potential for a false positive can lead to serious errors of interpretation, particularly when the suspect is identified through a "DNA dragnet" or database search, and that ignorance of the true rate of error creates an important element of uncertainty about the value of DNA evidence. PMID- 12570199 TI - Statistical analysis of barefoot impressions. AB - Comparison of the shapes of barefoot impressions from an individual with footprints or shoes linked to a crime may be useful as a means of including or excluding that individual as possibly being at the scene of a crime. The question of the distinguishability of a person's barefoot print arises frequently. This study indicates that measurements taken from the outlines of inked footprint impressions show a great degree of variability between donors and a great degree of similarity for multiple impressions taken from the same donor. The normality of the set of measurements on footprint outlines that we have selected for this study is confirmed. A statistical justification for the use of the product rule on individual statistical precisions is developed. PMID- 12570200 TI - Inhibition of bleach-induced luminol chemiluminescence. AB - The luminol chemiluminescence presumptive test for blood is based on the mild peroxidase activity of hemoglobin in basic peroxide solution. However, this test is subject to interference by strong oxidants, certain transition metal ions, and true peroxidases. This paper reports methods for reducing the interference caused by hypochlorite-containing bleaches. Amines such as 1,2-diaminoethane react rapidly with hypochlorite without interfering significantly with the hemoglobin catalyzed oxidation. Thus, addition of 0.1 mol/L 1,2-diaminoethane to a standard luminol-peroxide spray lead to almost complete inhibition of hypochlorite-induced chemiluminescence while satisfactory chemiluminescence was still observed from bloodstains. If time allows, an alternative method for reducing interference from hypochlorite bleach is to wait several days until the bloodstains have dried thoroughly, by which time the hypochlorite will have decomposed. PMID- 12570201 TI - Identification of pressure-sensitive adhesive polypropylene tape. AB - Identification of colorless, transparent, pressure-sensitive adhesive polypropylene tape (PP tape) was performed using infrared absorption spectrometry (IR) and pyrolysis gas chromatography mass spectrometry (Py-GC/MS) to determine the pressure-sensitive adhesive. Twenty different products of colorless, transparent PP tapes were examined in this study, and the results of analysis of IR spectra and Py-GC/MS were classified into twelve groups. In addition, the tapes were classified into 14 groups on the basis of IR measurement of release agent present in the backside. The results indicate that colorless, transparent PP tapes can be distinguished in terms of manufacturer, thereby demonstrating that this method of identifying colorless, transparent PP tape is effective. Moreover, the method was applied to the analysis of an actual forensic sample. PMID- 12570202 TI - Using resolution calculations to assess changes in capillary electrophoresis run parameters. AB - Capillary electrophoresis is widely used in the forensic community for the analysis of Short Tandem Repeat DNA. The CE system used in most forensic laboratories allows the user to modify standard operational protocols to accommodate some samples that fall outside of interpretational guidelines. We have made operational changes and monitored system resolution and the ability of the software to identify alleles as a result of these modifications. Increased amount of amplified product in the injection preparation, lengthened injection times or repeated injections of the same sample were all investigated along with variations in run voltages. Certain protocol modifications offer some advantages to the analysis of STRs and may allow the analyst to avoid extraction of additional portions of crime scene samples to achieve interpretable results. PMID- 12570203 TI - Characterization of a novel dimorphism in the 5' flanking region of the short tandem repeat (STR) locus, c-fes/fps (FES). AB - The FES short tandem repeat (STR) locus contains seven to 14 repeats of the tetranucleotide sequence ATTT. A novel 10 base pair dimorphism in the 5' flanking region of the FES locus was characterized in four broad populations: African American, Hispanic, Caucasian, and Asian. The absence of the 10 base pair sequence, or (-) allele, was closely linked to FES STR alleles with 10 or fewer repeats. The presence of the 10 base pair sequence, or (+) allele, was closely linked to FES STR alleles with 12 or more repeats. The (-) and (+) alleles occurred equally often in FES STR allele 11. The nucleotide sequence (5' GGCTGTTTTG-3') of the (+) allele, located 179 base pairs upstream of the FES STR, was determined to be consistent within and among the four populations. Statistical and sequence analysis confirmed the linkage between the two polymorphic sites. The results indicate that the exclusion rate of the FES locus is increased, above that for the STR alone, when both polymorphic characteristics are considered. PMID- 12570204 TI - Validation of cytochrome b sequence analysis as a method of species identification. AB - One of the stages of dealing with biological material submitted to forensic laboratories is species identification. The aim of the present work was to validate and assess the possibility of applying sequence analysis of the region coding cytochrome b as a method of species identification in the field of forensic science. DNA originating from individuals from major phyla of vertebrates was isolated by the organic method from various specimens. Extracted DNA was subjected to PCR and direct cycle sequencing using a universal pair of primers. The validation process, performed according to TWGDAM recommendations, revealed that the technique is a very sensitive and reliable method of species identification allowing analysis of tiny amounts of material and also degraded material, and can be useful in the field of forensic genetics. The case example presented here, concerning the determination of species origin of biological evidence collected from fatal road accident, confirms that analysis can be carried out even when there is no reference sample, and the sequences obtained can be assessed through analysis of their similarity to sequences for cytochrome b present in DNA databases. PMID- 12570205 TI - Evaluation of the DNA stability of forensic markers used in betel-quid chewers' oral swab samples and oral cancerous specimens: implications for forensic application. AB - Chewed betel-quid (BQ) residues are often considered vital biological evidence at crime scenes, since the human DNA extracted from the residues is actually from buccal epithelial cells and can be associated with suspects. BQ-chewing is also a risk factor for oral diseases and/or cancers. Archived medical oral-specimens can be used to identify specific individuals under adverse conditions, although STR markers are known to be unstable in various tumor tissues. This study evaluates the DNA stability of forensic marker systems in BQ-chewers' oral epithelial cells, and in archived clinical specimens of oral cancer patients. The genotypes of oral and paired peripheral blood samples in 200 subjects were compared, using the commercialized typing systems of HLA-DQA1, PM (including LDLR, GYPA, HBGG, D7S8, and GC loci), and AmpFlSTR markers (including 9 STR loci and the Amelogenin gene). The 100 healthy BQ-chewers had consistent oral swab and paired blood sample genotypes analyzed withboth DQA1/PM and STR marker systems. In the 100 oral cancer patients, one discordant result at D7S8 was found in the 600DQA1/PM marker loci, and 25 allelic alterations with expansion or contraction were detected in the 900 STR loci. The findings herein suggest that when cancerous specimens were tested, the HLA-DQA1/PM system with point polymorphism appears more reliable than the STR system with length polymorphism. Our results also indicate that healthy BQ-chewers' oral cotton swabs containing buccal epithelial cells are useful for forensic purposes using the HLA-DQA1, PM, and STR marker systems. PMID- 12570206 TI - Development and validation of a multiplexed Y-chromosome STR genotyping system, Y PLEX 6, for forensic casework. AB - A Y-chromosome multiplex polymerase chain reaction (PCR) amplification kit, known as Y-PLEX 6, has been developed for use in human identification. The Y-PLEX 6 kit enables simultaneous amplification of six polymorphic short tandem repeat (STR) loci located on the non-recombinant region of the human Y-chromosome. These loci are: DYS393, DYS19, DYS38911, DYS390, DYS391, and DYS385. Our studies show that as little as 0.2 ng of template DNA can be used for analysis. The specificity of the amplification reaction enabled analysis of male DNA in a male:female DNA mixture at a ratio of 1:125. Among the six Y-STR loci, the maximum mean stutter percentage was 11.9 for allele at DYS38911 locus. Attempts at amplification of DNA from various animal sources revealed that the Y-PLEX 6 primers are human specific. Details of the development of the kit, generation and description of the allelic ladders, and validation of the multiplex PCR are presented. In addition, Y-STR allele and haplotype frequencies in three populations have been investigated. The data indicate that results obtained using the Y-PLEX 6 kit are robust, sensitive, and reliable and can be used in human forensic and male lineage identification cases. PMID- 12570207 TI - Molecular phenotyping of a trinucleotide repeat (D5S373) experimental conditions. AB - The aim of this study is to assess the utility of the STR D5S373 in human identification. PCR amplification and electrophoretic separation were optimized in order to achieve unambiguous phenotyping. We concluded that primer concentration and annealing temperature are the main factors affecting the specificity of PCR. In our population survey including three human major groups (Europe, Sub-Saharan Africa, and Asia), up to six alleles and six interalleles have been found ranging in size from 86 to 101 bp. The phenotypes were determined using horizontal polyacrylamide gel electrophoresis, a technique which has turned out to be suitable for separating fragments as close as 1 bp. In each population, the genotype frequencies conformed to the expectations of genetic equilibrium. Sequence studies were carried out to make the allele nomenclature fit to ISFH recommendations. Results from our population analysis of D5S373 show clear differences in allelic frequency patterns among the three major human groups examined. Human identification parameters estimated from our study are similar to those obtained for other STRs currently used in DNA testing. PMID- 12570208 TI - A simple method of DNA extraction and STR typing from urine samples using a commercially available DNA/RNA extraction kit. AB - We devised a simple DNA extraction procedure suitable for STR typing of urine sample. Use of a commercially available DNA/RNA extraction kit equipped with a silica-gel-based membrane made it possible to omit the recovery of urinary nucleated cells by sedimentation before the extraction. Successful genotyping of the TH01, HumTPO and multiplex STRs was achieved using aliquots of urine as small as 100 microL. Furthermore, application of this DNA extraction procedure to frozen urine samples provided STR allele results comparable to results obtained from fresh samples. Therefore, this extraction procedure is considered to be effective for STR typing of urine samples in both the frozen and aqueous state. Furthermore, addition of sodium azide to fresh urine samples prolonged their storage duration even at room temperature. PMID- 12570209 TI - An alternative method to screen for pepper spray residue. AB - A method was developed to screen for pepper spray residue using instruments and methods other than those techniques commonly employed to analyze chemical residue (i.e.. gas chromatography mass spectrometry-GCMS or liquid chromatography mass spectrometry-LCMS). The method employed gas chromatography (GC), thin layer chromatography (TLC), and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) to screen for dried pepper spray stains. Pepper sprays from nine different manufacturers were investigated. Capsaicin and dihydrocapsaicin were identified and unique IR reflectance spectra are presented. An additional five compounds were presumptively found. Results showed that a particular stain could be characterized as a pepper-based stain. PMID- 12570210 TI - A novel fluorescent quadruplex STR typing system and the allele frequency distributions in a Thai population. AB - We have previously reported a new triplex amplification and typing system by silver staining for three short tandem repeat (STR) loci, 9q2h2 (D2S3020), D15S233, and D14S299 without "microvariant" alleles such as .1, .2, and, .3 alleles in the Japanese population. In the present study, we established a new quadruplex system with an additional locus D7S809 using primer sets labeled with fluorescent multi-color dyes. Using this system, we genotyped 183 Thai people, found only one "microvariant" allele (allele 20.2) at D7S809, and calculated allele frequencies and some statistical properties at these four STR loci. From these allele frequencies at four STR loci, we performed three statistical analyses including a homozygosity test, a likelihood ratio test, and an exact test for Hardy-Weinberg equilibrium (HWE). Deviations from HWE (p < 0.05) were observed only in the two tests at the locus D7S809. In the present study, we compared the allele frequencies at these four loci in the Thai population to those in the Japanese population described previously. Consequently, all observed heterozygosities and power of discrimination (PD) at those loci in the Thai population were higher than 0.8 and 0.9, respectively, and all statistical values for discriminating power in the Thai population were slightly higher than those in the Japanese population. The combined paternity exclusion rate (combined PE) in the Thai population (0.978) was almost the same as that in the Japanese population (0.971). Therefore, this novel PCR amplification and typing system for four STR loci would be a convenient and informative DNA profiling system in the forensic field. PMID- 12570211 TI - Stable-isotope fingerprints of biological agents as forensic tools. AB - Naturally occurring stable isotopes of light elements in chemical and biological agents may possess unique "stable-isotope fingerprints" depending on their sources and manufacturing processes. To test this hypothesis, two strains of bacteria (Bacillus globigii and Erwinia agglomerans) were grown under controlled laboratory conditions. We observed that cultured bacteria cells faithfully inherited the isotopic composition (hydrogen, carbon, and nitrogen) of media waters and substrates in predictable manners in terms of bacterial metabolism and that even bacterial cells of the same strain, which grew in media water and substrates of different isotopic compositions, have readily distinguishable isotopic signatures. These "stable-isotopic fingerprints" of chemical and biological agents can be used as forensic tools in the event of biochemical terrorist attacks. PMID- 12570212 TI - DNA profiling of azoospermic semen samples from vasectomized males by using Y PLEX 6 amplification kit. AB - Post-vasectomized azoospermic semen samples (N = 6) were analyzed for short tandem repeats (STR) on the Y-chromosome by using Y-PLEX 6 and the 310 Genetic Analyzer. We have observed a wide variation in the yield of extracted DNA from 12.5-1,000 ng. This variation was attributed to the number of epithelial and/or white blood cells that are present in these azoospermic samples. DNA profiles of these vasectomized males were obtained for all six Y-STR loci, namely DYS393, DYS 19, DYS389II, DYS390, DYS391, and DYS385 amplified by using the Y-PLEX 6. PMID- 12570213 TI - Preferential extraction of hydrocarbons from fire debris samples by solid phase microextraction. AB - Headspace analysis by extraction/GC-MS is a common method of detecting volatile hydrocarbon accelerants in fire debris samples. Solid-phase microextraction was tested to determine if there is selective extraction of chemically distinct compounds. It was found that both the polydimethylsiloxane (PDMS) and Carboxen/PDMS solid phase microextraction fibers show preferential extraction of aliphatic or aromatic compounds from the headspace depending on fiber type and temperature. The Carboxen/PDMS fiber type showed particular (although not exclusive) selectivity for extraction of aromatic hydrocarbons. Other experimental considerations of SPME are noted. PMID- 12570214 TI - Detection of fetal DNA in a cell pellet after centrifugation of mountant. AB - In order to obtain fetal cells (e.g., for paternity cases) after abortion, the centrifugation of mountant (in our case formalin) may be tried when the DNA examination of the fixed tissue itself gives limited or no (or not enough) information. The fixed tissue was microscopically negative for fetal cells and gave no satisfactory results when examined for DNA. Centrifugation of approximately 50 mL of reddish colored formalin resulted in a cell pellet that was examined for DNA, which gave enough information to confirm a case of sexual abuse. PMID- 12570215 TI - 'The story of Abraham, Isaac and Jacob" or "am I my brother's keeper?". AB - Presented is a case report of a violent sexual assault where the DNA profile obtained from an item of evidence was compared to a suspect's profile. The profiles did not match, but the sharing of such a large number of alleles raised the suspicion that perhaps the real perpetrator was a blood relative of the suspect. The investigators requested a sample from the suspect's brother, and a match was defined. In an era of technological breakthroughs in the field of forensic DNA analysis, the importance of the scientist's attention to the evidence presented in each case is stressed. PMID- 12570216 TI - Age estimation from teeth in children and adolescents. AB - The aim of this cross-sectional study was to evaluate the dental formulas according to age because of possible changes during the last decades. From these dental formulas, the median age of emergence for the permanent teeth was deduced. The study population corresponded to the 1 to 15 year-old children who had consulted either a pediatric dentist or orthodontist in the city of Nice (France). Data were collected from the 5,848 patients' charts that included an orthopantomography. This permitted the observation of emerged teeth and agenesis. Bar charts were used to indicate the dental formula according to age. There was no significant difference in the emergence pattern of both controlateral maxillary and mandibular teeth. Only the anterior tooth emergence significantly differed according to the maxillary. The lower central incisor was the lone tooth with a median age earlier than others; the remaining teeth had an age equivalent to those of previously estimates. PMID- 12570217 TI - Reliability and validity of eight dental age estimation methods for adults. AB - This paper evaluates the reliability and validity of eight published dental age estimation methods for adults that may aid in victim identification. Age was calculated on 20 Caucasian teeth of known age according to the methods of Kvaal (for in situ and extracted teeth), Solheim (for in situ and sectioned teeth), Lamendin (for extracted teeth), Johanson (for sectioned teeth) and Bang (for extracted and sectioned teeth) by one independent observer. For each method, mean age error and standard error were assessed as the measures of accuracy and precision. In addition, method simplicity, requirements for tooth preparation and the equipment necessary were assessed and recommendations given for forensic use in various situations. Methods for sectioned teeth gave more reliable results when compared to methods for intact teeth. PMID- 12570218 TI - The human ear: its role in forensic practice. AB - The human ear has much to offer investigators in relation to many areas of forensic practice including forensic pathology, anthropology, identification and facial reconstruction and yet its full potential is often overlooked. This review paper explores current knowledge related to the human ear drawing attention to those situations where the ear could play a valuable role in a forensic investigation. PMID- 12570219 TI - The happy land homicides: 87 deaths due to smoke inhalation. AB - We reviewed all 87 deaths from the Happy Land Social Club fire. All deaths were due to smoke inhalation. The carboxyhemoglobin (COHb) concentrations ranged from 37 to 93% with a mean of 76.5%. The vast majority (97%) of the decedents had a COHb concentration over 50%. Cyanide blood concentrations ranged from 0 to 5.5 mg/L with a mean of 2.2 mg/L. Nine decedents had no cyanide detected, and seven had cyanide concentrations of less than 1 mg/L. Fewer than one third of the decedents had thermal injuries, and most were partial thickness burns involving less than 20% body surface area. Ethanol was detected in 72% of decedents with a range of 0.01 to 0.29 g% and a mean blood concentration of 0.11 g%. Cocaine or cannabinoid use was identified in 9% of the decedents. All decedents were visually identified, and all had soot in the airway extending to the major bronchi. Carboxyhemoglobin concentrations corresponded well with deaths from smoke inhalation. Cyanide concentrations did not correspond with the extent of smoke inhalation, and the role of cyanide in contributing to these deaths is doubtful. Hydrogen chloride inhalation, as evidenced by comparison of the pH of tracheal mucosa to controls, was not a factor. PMID- 12570220 TI - Schizophrenia as a cause of death. AB - Schizophrenia is a chronic disorder that is associated with increased mortality. Although traumatic deaths account for most of this increase, there is also an increased rate of natural deaths in this population. Altered autonomic physiology in this group might contribute to death. To determine if there are schizophrenics in whom, after a complete autopsy, no recognizable cause of death other than schizophrenia is established, the records of the Office of Chief Medical Examiner of the City of New York were reviewed for deaths associated with schizophrenia and a natural manner of death. Six such decedents were identified, and their histories and autopsy results are described. We believe that schizophrenia per se is a potentially lethal disorder. Autonomic irregularities and their interactions with psychotropic drugs deserve further attention. PMID- 12570221 TI - Taphonomic mycota: fungi with forensic potential. AB - Forensic archaeologists and criminal investigators employ many different techniques for the location, recovery, and analysis of clandestine graves. Many of these techniques are based upon the premise that a grave is an anomaly and therefore differs physically, biologically, or chemically from its surroundings. The work reviewed in this communication demonstrates how and why field mycology might provide a further tool towards the investigation of scenes of crime concealed in forest ecosystems. The fruiting structures of certain fungi, the ammonia and the postputrefaction fungi, have been recorded repeatedly in association with decomposed mammalian cadavers in disparate regions of the world. The ecology and physiology of these fungi are reviewed briefly with a view to their potential as a forensic tool. This application of mycology is at an interface with forensic archaeology and forensic taphonomy and may provide a means to detect graves and has the potential to estimate postburial interval. PMID- 12570222 TI - Mechanisms of unexpected death in tuberous sclerosis. AB - Tuberous sclerosis complex is a protean autosomal dominant disorder characterized by multifocal tissue lesions arising from defects in cellular migration, proliferation, and differentiation. It has an association with sudden death. In the current study, review of all cases of sudden death due to tuberous sclerosis was undertaken at the Forensic Science Centre in Adelaide, Australia from 1991 to 2001, in addition to an analysis of cases from the literature. There were two local cases where unexpected death had occurred in individuals with known tuberous sclerosis, involving a 31-year-old male (epilepsy), and a 24-year-old female (massive hemorrhage into a renal angiomyolipoma). Fatal mechanisms in cases of tuberous sclerosis may be associated with underlying cardiovascular, renal and cerebral abnormalities. Sudden death may be due to cardiac arrhythmia, epilepsy, and intra-tumoral hemorrhage with additional complications including cardiac outflow obstruction, obstructive hydrocephalus, aneurysm rupture, and spontaneous pneumothorax. An awareness of the highly variable tissue manifestations of tuberous sclerosis and the mechanisms that may be responsible for death is necessary to establish correctly the diagnosis in occult cases (possibly with molecular confirmation), and to chart accurately organ changes in individuals with established disease. PMID- 12570223 TI - Spontaneous cerebellar hemorrhage in children. AB - Spontaneous cerebellar hemorrhages are a rare but often fatal occurrence in children. Although there are several predisposing factors such as blood dyscrasias or astrocytomas, the most common cause of cerebellar hemorrhage in an otherwise healthy child is the rupture of a vascular malformation. We reviewed the files of the Office of Chief Medical Examiner of the City of New York and found four such instances over a period of less than two years. We present these here and outline the approach the forensic pathologist should take in evaluating such deaths. PMID- 12570224 TI - Stun gun injuries in the abuse and death of a seven-month-old infant. AB - The author autopsied a seven-month-old infant who was shocked repeatedly with a stun gun by his foster mother, in an attempt by the foster mother to get the infant to stop crying. The stun gun injuries were round, well-circumscribed, erythematous macular lesions, which were found in pairs. The lesions were 2 in. apart, and were found to match the distance between the electrodes of the stun gun found in the foster mother's purse. Based on the electrical output of the stun gun, the small size of the infant, location of stun gun discharge, and the decreased resistance of the infant's skin, it can be concluded that the stun gun injury is responsible for the infant's death. PMID- 12570225 TI - Fatal parvovirus B19 myocarditis in an 8-year-old boy. AB - A report is given on an 8-year-old boy who suddenly and unexpected died. Autopsy findings point to acute heart failure. Microscopic examination of the heart showed increased interstitial and perivasal fibrosis and myocarditis with macrophage infiltration. Polymerase chain reaction (PCR) analysis for parvovirus B19 was positive in heart samples and in the spleen. Immunostaining for parvoviral surface antigens was negative. Although the virus does not appear to have infected the cardiomyocytes, we speculate that myocarditis arose from immunological cross-reaction to epitopes shared between the virus and the myocardium. PMID- 12570226 TI - Neuroleptic-induced akathisia and violence: a review. AB - Surprisingly, the association of neuroleptic-induced akathisia and aggressive behavior was not formally recognized until nearly two and one-half decades of antipsychotic prescribing had passed. Using a search of the anglophonic literature, this phenomenon is reviewed. Advances in psychopharmacology have reduced neuroleptic-induced akathisia and hold promise to eliminate it altogether. Nonetheless, important clinical and forensic aspects of neuroleptic induced akathisia and aggression remain and are explored. PMID- 12570227 TI - Attempted suicide by cop: a case study of traumatic brain injury and the insanity defense. AB - Over the years, there have been a number of well-publicized incidents involving persons who seemingly maneuver police officers into shooting them. Such cases, while relatively rare compared with most forms of violence, nevertheless pose difficult challenges to law enforcement agencies. Relatively little is known about persons who engage in suicide by cop incidents. To our knowledge, there has been no published indepth research on instigators of suicide by cop who survived. In this paper, we present a case study of an individual who engaged in and lived through three separate "attempted suicide by cop" incidents. After describing relevant history as well as events of the most recent incident, we compare the case with the extant literature on suicide by cop and analyze commonalities and differences. Finally, we examine the legal considerations involved, with particular attention devoted to the role the individual's traumatic brain injury played in applying the insanity defense. PMID- 12570228 TI - Testing for GHB in hair by GC/MS/MS after a single exposure. Application to document sexual assault. AB - Gamma-hydroxybutyric acid, or GHB, is a substance naturally present within mammal species. Properties of neurotransmitter or neuromodulator are generally given to this substance. GHB is therapeutically used as an anesthetic, but can be used for criminal offenses (date-rape drug). It appears that the window of detection of GHB is very short in both blood and urine, and therefore its presence is very difficult to prove after a rape case. In order to document single exposure, we investigated the use of hair. Hair was collected one month after the allegated event in order to sample the corresponding period after regular growing. After rapid (2 min) decontamination with dichloromethane, the hair shaft was cut into 3 mm segments. They were overnight incubated in 0.01 N NaOH in the presence of GHB d6, followed by neutralization and extraction in ethyl acetate under acidic conditions. GHB (precursor ion m/z 233, product ions m/z 147 and 148) was tested by GC/MS/MS (Finnigan TSQ 700) after derivatization with BSTFA + 1% TMCS. Physiological concentrations (n = 24) were in the range 0.5 to 12.0 ng/mg, with no influence due to hair color. No variation of concentrations was observed along the hair shaft in controlled subjects, except for the proximal segment, due to an incorporation through sweat. This demonstrates that endogenous levels for each single subject are constant during hair growth. A controlled human administration of 25 mg/kg to a volunteer demonstrated that a single exposure to GHB is detectable in hair after segmentation. In a case of rape under influence, a clear increase of the corresponding segment (about 2.4 ng/mg) in time was observed, in comparison with the other segments (0.6 to 0.8 ng/mg). This study demonstrates that a single exposure to GHB in a case of sexual assault can be documented by hair analysis when collected about one month after the crime. PMID- 12570229 TI - Basque country autochthonous population data on D8S1179, D21S11, D18S51 and D16S539 loci. PMID- 12570230 TI - Angola population data with AmpFlSTR profiler plus. PMID- 12570231 TI - Haplotype distribution of nine Y-chromosome STR-loci in two northern Spanish populations (Asturias and Aragon). PMID- 12570232 TI - Allele frequencies for two STR loci D21S1436, D21S2052 in Chinese population. PMID- 12570233 TI - Allele frequencies for D1S80 (pMCT118) locus in some East European populations. PMID- 12570234 TI - Allele frequencies for six STR loci (D5S818, D7S820, D8S1179, D13S317, D16S539, FGA) in a population sample of North Italy. PMID- 12570235 TI - Autosomal microsatellite profile of three socially diverse ethnic Tamil populations of India. PMID- 12570236 TI - Y-chromosomal STR haplotypes at the loci DYS19, DYS385, DYS38911, DYS390, DYS391, and DYS393 in Marmara Region of Turkey. PMID- 12570237 TI - Turkish population data on the CTTV STR loci. PMID- 12570238 TI - Allele frequencies for two STR loci D11S1977 and D22S444 in Chinese population. PMID- 12570239 TI - Gene frequencies for three hypervariable DNA loci in a Chilean population of mixed ancestry. PMID- 12570240 TI - Haplotype diversity of two Y-chromosomal SNPs in Chinese populations. PMID- 12570241 TI - Distribution of the D17S5 alleles in South-East Spain (Murcia Region). PMID- 12570242 TI - Population data of Ecuador for fifteen STR loci (PowerPlex 16). PMID- 12570243 TI - Mitochondrial DNA hypervariable region I and II sequence polymorphism in the Dravidian linguistic group of India. PMID- 12570244 TI - Allele frequencies of nine STR loci in Chinese She ethnic group from Zhejiang, China. PMID- 12570245 TI - Allele frequencies and statistical parameters for eight STR loci in Cordoba (Argentina) population. PMID- 12570246 TI - French allele frequencies and haplotypes of nine Y-specific STRs. PMID- 12570247 TI - Allele frequency of eight Y-chromosome STR loci in oriya population of India. PMID- 12570249 TI - Commentary on: Fregean CJ, Vanstone H, Borys S, McLean D, et al. AmpFlSTR Profile Plus and AmpFlSTR Cofiler analysis of tissues stored in GenoFix, a new tissue preservative solution for mass disaster DNA identification. J Forensic Sci 2001;46(5):1180-90. PMID- 12570248 TI - Paraguayan population data on the fifteen STR loci included in the PowerPlex 16 kit. PMID- 12570251 TI - Halo polarization profiles and sampled ice crystals: observations and interpretation. AB - Simultaneous two-wavelength polarization and radiance distributions have been obtained for 22 degrees parhelia in four Antarctic ice-crystal swarms that extended to ground level. Samples of crystals that produced these parhelia were collected and replicated. The wavelength dependence of the width of the halo polarization peak agrees with Fraunhofer diffraction theory, indicating that the broadening of the halos is caused primarily by diffraction. However, the observed broadening is much more than predicted from the size distribution of the replicated crystals. From one halo display to the other, the ratio of observed/predicted broadening is erratic, suggesting size-dependent collection efficiency in the sampling. This would imply that, for South Pole conditions, halo polarimetry (or even photometry) is a more reliable method for crystal size determination than actual sampling. It also implies that shapes of the sampled crystals need not necessarily be representative for the shapes of the halo-making crystals in the swarm. Our previous hypothesis [Appl. Opt. 33,4569 (1994)], that a spread of interfacial angles is the dominating cause of halo broadening, has proved untenable. PMID- 12570252 TI - Symmetry in halo displays and symmetry in halo-making crystals. AB - The relation between the symmetry in halo displays and crystal symmetry is investigated for halo displays that are generated by ensembles of crystals. It is found that, regardless of the symmetry of the constituent crystals, such displays are always left-right (L-R) symmetric if the crystals are formed from the surrounding vapor. L-R symmetry of a halo display implies here that the cross sections for formation of a halo arc on the left-hand side of the solar vertical and its right-hand side mirror image are equal. This property leaves room for two types of halo display only: a full symmetric one (mmm-symmetric), and a partial symmetric one (mm2-symmetric) in which halo constituents lack their counterparts on the other side of the parhelic circle. A partial symmetric display can occur only for point halos. Its occurrence implies that a number of symmetry elements are not present in the shape of the halo-making crystals. These elements are a center of inversion, any rotatory-inversion axis that is parallel to the crystal spin axis P, a mirror plane perpendicular to the P axis, and a twofold rotation axis perpendicular to the P axis. A simple conceptual method is presented to reconstruct possible shapes of the halo-generating crystals from the halos in the display. The method is illustrated in two examples. Halos that may occur on the Saturnian satellite Titan are discussed. The possibilities for the Huygens probe to detect these halos during its descent through the Titan clouds in 2005 are detailed. PMID- 12570253 TI - Midlatitude cirrus cloud climatology from the Facility for Atmospheric Remote Sensing. IV. Optical displays. AB - In this fourth of a series of papers that describe long-term cloud research at the Facility for Atmospheric Remote Sensing at Salt Lake City, Utah, an approximately 10-year record of polarization lidar and photographic observations is analyzed to characterize the occurrence of optical displays in our local varieties of midlatitude cirrus clouds. The frequencies of occurrence of various types of halo, arc, and corona displays are evaluated according to their appearance and longevity over nominal 1-h observation periods and to the meteorological source of the cirrus. We find that complex halo-arc displays are rare at our locale and that even the so-called common 22 degree halo occurs infrequently as a complete long-lived ring. For example, only approximately 6% of the 1561-h daytime cirrus periods have bright and prolonged 22 degree halos, although a total of 37.3% have some indications of this halo, even if they are brief and fragmentary. Other fairly frequent features are the 22 degree upper tangent arc (8.6%), 22 degree parhelia (8.5%), and solar corona (7.2%). Of the optical displays observed, 83.6% are refraction based, only 1.9% are due to reflection phenomena, and a surprising 15.4% are caused by diffraction. Complex halo-arc displays are disproportionally associated with cirrus formed in tropical or subtropical airflow and also contain more horizontally oriented planar ice crystals. Lidar linear depolarization ratios from a subset of vivid displays show significant differences between halo- and the corona-producing cirrus, reflecting the effects of particle shape. Halos are associated with relatively warm cirrus that contain randomly and horizontally oriented planar ice crystals, whereas the colder corona cirrus produce much stronger depolarization from crystals too small to be uniformly oriented. Comparisons are made with available information from other locales, and we attempt to explain the geographical differences in terms of basic cirrus cloud processes. PMID- 12570254 TI - Comparison of modeled and observed astronomical refraction of the setting sun. AB - In this study a ray-tracing model that uses atmospheric data from VIZ and Vaisala RS80 rawinsondes is compared with the observed astronomical refraction presented by the setting Sun as seen from Stony Plain, Alberta, Canada. Photogrammetric measurements taken from photographs of the setting Sun show good agreement with the model for the 14 and 22 December 1998 sunsets. The poorer model results for the 8 December sunset appear to be caused by an obsolete and possibly defective VIZ rawinsonde. The results suggest that the ray-tracing model can produce improved refraction values when compared with the Pulkovo tables [Pulkovo Observatory, Refraction Tables of the Pulkovo Observatory, 5th ed. (Nauka, Leningrad, 1985)]. However, they also indicate that the inverse solution (i.e., extracting the temperature profile from refraction measurements) may produce no improvement on U.S. Standard Atmosphere adjusted to the surface conditions. PMID- 12570255 TI - Ray tracing and refraction in the modified US1976 atmosphere. AB - A new and flexible ray-tracing procedure for calculating astronomical refraction is outlined and applied to the US1976 standard atmosphere. This atmosphere is generalized to allow for a free choice of the temperature and pressure at sea level, and in this form it has been named the modified US1976 (MUSA76) atmosphere. Analytical expressions and numerical procedures are presented for calculating dry-air refractions and for the water-vapor correction. Results for all apparent altitudes are presented and compared with The Star Almanac for Land Surveyors (1951), The Nautical Almanac (1958), and the Pulkovo tables (Refraction Tables of the Pulkovo Observatory, 1985). Dependences on sea-level pressure, temperature, and temperature gradient and on humidity are discussed. PMID- 12570256 TI - Novaya Zemlya effect and sunsets. AB - Systematics of the Novaya Zemlya (NZ) effect are discussed in the context of sunsets. We distinguish full mirages, exhibiting oscillatory light paths and their onsets, the subcritical mirages. Ray-tracing examples and sequences of solar images are shown. We discuss two historical observations by Fridtjof Nansen and by Vivian Fuchs, and we report a recent South Pole observation of the NZ effect for the Moon. PMID- 12570257 TI - Gerrit de Veer's true and perfect description of the Novaya Zemlya effect, 24-27 January 1597. AB - The first recordings of the Novaya Zemlya (NZ) effect were made during Willem Barents' third Arctic expedition. Ray-tracing analyses of the three key observations, on 24-27 January 1597, show that all the reported details can be explained by adopting one common and realistic type of temperature inversion. In particular, the Moon-Jupiter conjunction could have been visible over the central mountain ridge of the island. We show that the NZ effect distorts the relative positions of Jupiter and the Moon in such a way that the looked-for fingerprint of the conjunction occurred almost 2 h after the true conjunction. The quoted direction for the apparent Moon-Jupiter conjunction is then found to be accurate to within 1 degree. This delay of the apparent conjunction largely explains the error of 29 degrees in their longitude determination. The truthfulness of these observations, debated for four centuries, now appears to be beyond doubt. PMID- 12570258 TI - Bright superior mirages. AB - Superior mirages of unusual brightness are occasionally observed. Two such cases, photographed over the frozen surface of Lake Winnipeg, Canada, are documented. Visually, these mirages appear as featureless bright barriers far out on the lake. They are just images of the lake ice, yet the luminance in one case was 2.5 times (in the other, 1.7 times) the luminance of the ice surface in front of the mirage. The mirage itself can be modeled by means of a conduction inversion, but a proper explanation of the brightness is not yet available. PMID- 12570259 TI - Halo and mirage demonstrations in atmospheric optics. AB - Some laboratory demonstrations on atmospheric optics are presented. The focus is on dispersion effects in mirages, lateral mirages, and inferior mirages produced with small hot plates. We also show a demonstration of the upper-tangent-arc halo, produced with a hexagonal prism, rotating about two axes. PMID- 12570260 TI - Imaging polarimetry of the rainbow. AB - Using imaging polarimetry, we measured the polarization patterns of a rainbow on the shore of the Finnish town of Oulu in July 2001. We present here high resolution color-coded maps of the spatial distributions of the degree and angle of linear polarization of the rainbow in the red (650 +/- 30 nm), green (550 +/- 30 nm), and blue (450 +/- 30 nm) ranges of the spectrum. The measured polarization characteristics of the investigated rainbow support earlier theoretical and computational results and are in accordance with previous qualitative observations. To our knowledge, this is the first imaging polarimetric study of rainbow polarization. PMID- 12570261 TI - Experimental observation of total-internal-reflection rainbows. AB - A new class of rainbows is created when a droplet is illuminated from the inside by a point light source. The position of the rainbow depends on both the index of refraction of the droplet and the position of the light source, and the rainbow vanishes when the point source is too close to the center of the droplet. Here we experimentally measure the position of the transmission and one-internal reflection total-internal-reflection rainbows, and the standard (primary) rainbow, as a function of light-source position. PMID- 12570262 TI - Observation of the enhanced backscattering of light by the end of a tilted dielectric cylinder owing to the caustic merging transition. AB - The scattering of light by obliquely illuminated circular dielectric cylinders was previously demonstrated to be enhanced by a merger of Airy caustics at a critical tilt angle. [Appl. Opt. 37, 1534 (1998)]. A related enhancement is demonstrated here for backward and near-backward scattering for cylinders cut with a flat end perpendicular to the cylinder's axis. It is expected that merged caustics will enhance the backscattering by clouds of randomly oriented circular cylinders that have appropriately flat ends. PMID- 12570263 TI - Analysis of the shadow-sausage effect caustic. AB - We analyze the optical caustic produced by light refracted at the curved meniscus surrounding a cylindrical rod standing partially out of a liquid-filled container. When the rod is tilted from the vertical or when light is diagonally incident, the caustic is a four-cusped astroid with two of its cusps obscured by the rod's shadow. If a portion of the flat end of the rod is raised above the water level, the caustic evolves into a pattern of five interlocking cusps. The five cusps result from symmetry breaking of a three-cusped surface perturbation caustic. PMID- 12570264 TI - Simulating glories and cloudbows in color. AB - Glories and cloudbows are simulated in color by use of the Mie scattering theory of light upwelling from small-droplet clouds of finite optical thickness embedded in a Rayleigh scattering atmosphere. Glories are generally more distinct for clouds of droplets of as much as approximately 10 microm in radius. As droplet radius increases, the glory shrinks and becomes less prominent, whereas the cloudbow becomes more distinct and eventually colorful. Cloudbows typically consist of a broad, almost white band with a slightly orange outer edge and a dark inner band. Multiple light and dark bands that are related to supernumerary rainbows first appear inside the cloudbow as droplet radius increases above approximately 10 microm and gradually become more prominent when all droplets are the same size. Bright glories with multiple rings and high color purity are simulated when all droplets are the same size and every light beam is scattered just once. Color purity decreases and outer rings fade as the range of droplet sizes widens and when skylight, reflected light from the ground or background, and multiply scattered light from the cloud are included. Consequently, the brightest and most colorful glories and bows are seen when the observer is near a cloud or a rain swath with optical thickness of approximately 0.25 that consists of uniform-sized drops and when a dark or shaded background lies a short distance behind the cloud. PMID- 12570265 TI - Simulation of rainbows, coronas, and glories by use of Mie theory. AB - Mie theory offers an exact solution to the problem of scattering of sunlight by spherical drops of water. Until recently, most applications of Mie theory to scattering of light were restricted to a single wavelength. Mie theory can now be used on modern personal computers to produce full-color simulations of atmospheric optical effects, such as rainbows, coronas, and glories. Comparison of such simulations with observations of natural glories and cloudbows is encouraging. PMID- 12570266 TI - Measuring and modeling twilight's purple light. AB - During many clear twilights, much of the solar sky is dominated by pastel purples. This purple light's red component has long been ascribed to transmission through and scattering by stratospheric dust and other aerosols. Clearly the vivid purples of post-volcanic twilights are related to increased stratospheric aerosol loading. Yet our time-series measurements of purple-light spectra, combined with radiative transfer modeling and satellite soundings, indicate that background stratospheric aerosols by themselves do not redden sunlight enough to cause the purple light's reds. Furthermore, scattering and extinction in both the troposphere and the stratosphere are needed to explain most purple lights. PMID- 12570267 TI - Color and luminance asymmetries in the clear sky. AB - A long-standing assumption about the clear sky is that its colors and luminances are distributed symmetrically about the principal plane. As useful as this approximation is, our digital-image analyses show that clear-sky color and luminance routinely depart perceptibly from exact symmetry. These analyses reconfirm our earlier measurements with narrow field-of-view spectroradiometers [J. Opt. Soc. Am. A 18, 1325 (2001)], and they do so with much higher temporal and angular resolution across the entire sky dome. PMID- 12570268 TI - Neutral points of skylight polarization observed during the total eclipse on 11 August 1999. AB - We report here on the observation of unpolarized (neutral) points in the sky during the total solar eclipse on 11 August 1999. Near the zenith a neutral point was observed at 450 nm at two different points of time during totality. Around this celestial point the distribution of the angle of polarization was heterogeneous: The electric field vectors on the one side were approximately perpendicular to those on the other side. At another moment of totality, near the zenith a local minimum of the degree of linear polarization occurred at 550 nm. Near the antisolar meridian, at a low elevation another two neutral points occurred at 450 nm at a certain moment during totality. Approximately at the position of these neutral points, at another moment of totality a local minimum of the degree of polarization occurred at 550 nm, whereas at 450 nm a neutral point was observed, around which the angle-of-polarization pattern was homogeneous: The electric field vectors were approximately horizontal on both sides of the neutral point. PMID- 12570269 TI - Coronas and iridescence in mountain wave clouds. AB - We use Fraunhofer diffraction theory and meterological data to determine the nature of cloud-particle distributions and the mean particle sizes required for interpreting photographs of coronas and iridescence in mountain wave clouds. Traditional descriptions of coronas and iridescence usually explain these optical phenomena as diffraction by droplets of liquid water. Our analysis shows that the photographed displays have mean particle sizes from 7.6 to 24.3 microm, with over half the cases requiring diffraction by small (approximatley 20 microm) quasispherical ice particles rather than liquid water droplets. Previous documentation of coronas produced by ice particles are limited to observations in cirrus clouds that appear to be composed of small ice crystals, whereas our observations suggest that coronas and iridescence quite often can be created by tiny quasispherical ice particles that might be unique to mountain wave clouds. Furthermore, we see that the dominant colors in mountain wave-cloud coronas are red and blue, rather than the traditionally described red and green. PMID- 12570270 TI - Cirrus cloud iridescence: a rare case study. AB - On the evening of 25 November 1998, a cirrus cloud revealing the pastel colors of the iridescence phenomenon was photographed and studied by a polarization lidar system at the University of Utah Facility for Atmospheric Remote Sensing (FARS). The diffraction of sunlight falling on relatively minute cloud particles, which display spatial gradients in size, is the cause of iridescence. According to the 14-year study of midlatitude cirrus clouds at FARS, cirrus rarely produce even poor iridescent patches, making this particularly long-lived and vivid occurrence unique. In this unusually high (13.2-14.4-km) and cold (-69.7 degrees to -75.5 degrees) tropopause-topped cirrus cloud, iridescence was noted from approximately 6.0 degrees to approximately 13.5 degrees from the Sun. On the basis of simple diffraction theory, this indicates the presence of particles of 2.5-5.5-microm effective diameter. The linear depolarization ratios of delta = 0.5 measured by the lidar verify that the cloud particles were nonspherical ice crystals. The demonstration that ice clouds can generate iridescence has led to the conclusion that iridescence is rarely seen in midlatitude cirrus clouds because populations of such small particles do not exist for long in the presence of the relatively high water-vapor supersaturations needed for ice-particle nucleation. PMID- 12570271 TI - Solar aureoles caused by dust, smoke, and haze. AB - The forward scattering of sunlight by atmospheric aerosols causes a bright glow to appear around the Sun. This phenomenon, the simplest manifestation of the solar corona, is called the solar aureole. Simple methods can be used to photograph the solar aureole with conventional and digital cameras. Aureole images permit both a visually qualitative and an analytically quantitative comparison of aureoles caused by dust, smoke, haze, pollen, and other aerosols. Many hundreds of aureole photographs have been made at Geronimo Creek Observatory in Texas, including a regular time series since September 1998. These images, and measurements extracted from them, provide an important supplement to studies of atmospheric aerosols. PMID- 12570272 TI - Simulating coronas in color. AB - Coronas are simulated in color by use of the Mie scattering theory of light by small droplets through clouds of finite optical thickness embedded in a Rayleigh scattering atmosphere. The primary factors that affect color, visibility, and number of rings of coronas are droplet size, width of the size distribution, and cloud optical thickness. The color sequence of coronas and iridescence varies when the droplet radius is smaller than approximately 6-microm. As radius increases to approximately 3.5 microm, new color bands appear at the center of the corona and fade as they move outward. As the radius continues to increase to approximately 6 microm, successively more inner rings become fixed in the manner described by classical diffraction theory, while outer rings continue their outward migration. Wave clouds or rippled cloud segments produce the brightest and most vivid multiple ringed coronas and iridescence because their integrated dropsize distributions along sunbeams are much narrower than in convective or stratiform clouds. The visibility of coronas and the appearance of the background sky vary with cloud optical depth tau. First the corona becomes visible as a white aureole in a blue sky when tau approximately 0.001. Color purity then rapidly increases to an almost flat maximum in the range 0.05 < or = tau < or = 0.5 and then decreases, so coronas are almost completely washed out by a bright gray background when tau > or = 4. PMID- 12570273 TI - Evaluation of a one-dimensional cloud model for yellow and green thunderstorms. AB - Many observers have reported observations of green light emanating from severe thunderstorms in the midwestern United States. Spectral measurements have demonstrated that the dominant wavelength of the light is in the green portion of the visible spectrum and that this is not just a subjective impression. According to the theory proposed by Bohren and Fraser [Bull. Am. Meteorol. Soc. 74, 2185 (1993)], two effects combine to produce green light from thunderstorms. First, incident solar radiation is reddened by selective scattering by air molecules and particles in the atmosphere before it enters the cloud. Second, the radiation that passes through an optically thick cloud is attenuated in the longer wavelengths because of selective absorption by liquid water. Model calculations indicate that realizable combinations of mean drop diameters, mean liquid-water contents, and cloud thicknesses can satisfy the conditions required for shifting the dominant wavelength of the incident solar radiation to green. PMID- 12570274 TI - Polarization-based vision through haze. AB - We present an approach for easily removing the effects of haze from passively acquired images. Our approach is based on the fact that usually the natural illuminating light scattered by atmospheric particles (airlight) is partially polarized. Optical filtering alone cannot remove the haze effects, except in restricted situations. Our method, however, stems from physics-based analysis that works under a wide range of atmospheric and viewing conditions, even if the polarization is low. The approach does not rely on specific scattering models such as Rayleigh scattering and does not rely on the knowledge of illumination directions. It can be used with as few as two images taken through a polarizer at different orientations. As a byproduct, the method yields a range map of the scene, which enables scene rendering as if imaged from different viewpoints. It also yields information about the atmospheric particles. We present experimental results of complete dehazing of outdoor scenes, in far-from-ideal conditions for polarization filtering. We obtain a great improvement of scene contrast and correction of color. PMID- 12570275 TI - Flow cytometric determination of size and complex refractive index for marine particles: comparison with independent and bulk estimates. AB - We advance a method to determine the diameter D and the complex refractive index (n + n'i) of marine particles from flow cytometric measurements of forward scattering, side scattering, and chlorophyll fluorescence combined with Mie theory. To understand better the application of Mie theory with its assumptions to flow cytometry (FCM) measurements of phytoplankton cells, we evaluate our flow cytometric-Mie (FCM-Mie) method by comparing results from a variety of phytoplankton cultures with independent estimates of cell D and with estimates of n and n' from the inversion of bulk measurements. Cell D initially estimated from the FCM-Mie method is lower than independent estimates, and n and n' are generally higher than bulk estimates. These differences reflect lower forward scattering and higher side scattering for single-cell measurements than predicted by Mie theory. The application of empirical scattering corrections improves FCM Mie estimates of cell size, n, and n'; notably size is determined accurately for cells grown in both high- and low-light conditions, and n' is correlated with intracellular chlorophyll concentration. A comparison of results for phytoplankton and mineral particles suggests that differences in n between these particle types can be determined from FCM measurements. In application to natural mixtures of particles, eukaryotic pico/nanophytoplankton and Synechococcus have minimum mean values of n' in surface waters, and nonphytoplankton particles have higher values of n than phytoplankton at all depths. PMID- 12570277 TI - Foundations for low-loss fiber gradient-index lens pair coupling with the self imaging mechanism. AB - A fiber-optic collimator that emits a Gaussian beam with its beam waist at a certain distance after the exit face of the lens is labeled a self-imaging collimator. For such a collimator, the waist of the emitted Gaussian beam and its location are partly dependent on the properties of the gradient-index (GRIN) lens. Parameters for the self-imaging collimator are formulated in terms of the parameters of a GRIN lens (e.g., pitch, core refractive index, gradient index, length) and the optical wavelength. Next, by use of the Gaussian beam approximation, a general expression for the coupling power loss between two self imaging-type single-mode fiber (SMF) collimators is, for the first time to our knowledge, derived as a function of three types of misalignment, namely, separation, lateral offset, and angular tilt misalignment. A coupling experiment between two self-imaging collimators with changing separation distance is successfully performed and matches the proposed self-imaging mechanism coupling loss theory. In addition, using a prism, lateral offset, as well as angular tilt, misalignments are experimentally simulated for a two self-imaging collimator coupling condition by a single collimator reflective test geometry. Experimental results agree well with the proposed loss formulas for self-imaging GRIN lenses. Hence, for the first time to our knowledge, the mathematical foundations are laid for employing self-imaging-type fiber collimators in SMF-based free-space systems allowing optimal design for ultra-low-loss coupling. PMID- 12570276 TI - Pitfalls in atmospheric correction of ocean color imagery: how should aerosol optical properties be computed?: Comment. AB - The Sea-Viewing Wide Field-of-View Sensor (SeaWiFS) experience suggests that in most situations the aerosol models presently in use for atmospheric correction of ocean color imagery are sufficient for this task. It has been shown [Appl. Opt. 41, 412 (2002)] that the top-of-atmosphere reflectances computed for more realistic aerosol models differ from those computed for presently used models but have not shown that they will yield a better atmospheric correction, e.g., through direct application to ocean color imagery. Thus they provide no evidence that the presently used aerosol models are inadequate, or that their use is a pitfall in atmospheric correction. PMID- 12570278 TI - Analysis of transient-grating signals for reacting-flow applications. AB - Single-shot transient-grating measurements for thermometry in pressurized reacting flows are examined in the context of rapid digital signal processing. Simple approaches are discussed for temperature determination and rejection of unwanted signals in real-time measurement applications. Examples of temperature data in pressurized postflame gases are presented in the form of probability density functions (PDFs). Three contributions to the PDF half-widths are discussed. Analysis of phase-matching requirements indicates that beam steering as a result of density fluctuations affects the signal amplitude but not the grating period. Therefore, such stochastic beam deviations have little effect on the derived temperatures. Mode noise on the cw probe beam as well as linear light scattering are found to be insignificant in the frequency range of the observed transient-grating acoustic signature. Use of a single-mode laser for the pump beams is shown to enhance the signal intensity. PMID- 12570279 TI - Optical breakdown in fused silica and argon gas: application to Nd:YAG laser limiter. AB - A gas cell filed with argon gas under pressure is placed in a tightly focused laser beam to provide a limiter for laser pulses above a certain peak power, corresponding to the optical breakdown threshold for the creation of a laser induced plasma. Measurements of the threshold intensity as a function of argon gas pressure are given for a laser wavelength of 1.064 microm (Nd:YAG) and a pulse length of 6.4 ns. Threshold intensities for optical breakdown in fused silica were measured with the same optical system, enabling a relative comparison of breakdown thresholds, of interest for protecting fused-silica optical components in fiber-optic delivery systems for laser material processing applications. The threshold intensity was measured to 220 GW/cm2 in Ar at 1.0 x 10(5) N/m2 (1 atm), 80 GW/cm2 in Ar at 8.0 x 10(5) N/m2 (7.9 atm), and 55 GW/cm2 in fused silica. Even though the threshold in argon is higher than that in fused silica, the limiter will protect the optical components if the laser beam is focused to a tighter spot in the gas cell than at the input end of the fiber. PMID- 12570280 TI - Stable near-end solution of the lidar equation for clear atmospheres. AB - A stable variant of the near-end solution has been developed for inversion of lidar signals measured in clear atmospheres. The inversion is based on the use of reference values of the extinction coefficient obtained with a nephelometer at the lidar measurement site. The inversion method, based on a combination of the optical depth and boundary point solutions, is illustrated by simulated and experimental data. PMID- 12570281 TI - Far-infrared laser measurement of the refractive index of polypropylene. AB - The refractive index of polypropylene in the far infrared (FIR) is measured by means of a suitably modified laser of a FIR spectrometer. When thin polypropylene films of 12.7-microm nominal thickness are introduced in the optical cavity of a laser at the Brewster angle, the radiation ceases because of the change in the optical path of the laser beam. This change is measured from the displacement of one of the laser mirrors, which is necessary to restore the laser resonance. The refractive index of polypropylene is deduced from this measurement and from the film thickness, as obtained from an independent measurement based in pycnometry. The value obtained for the refractive index is 1.492(15) for the wavelengths between 118.834 and 251.140 microm, for a polypropylene film of 12.71(2)-microm thickness and 0.9049(7) g/cm3 density. PMID- 12570282 TI - Systematic review of the quality of randomized controlled trials for patellofemoral pain syndrome. AB - STUDY DESIGN: Systematic review of the literature. OBJECTIVES: To develop a grading scale to judge the quality of randomized clinical trials (RCTs) and conduct a systematic review of the published RCTs that assess nonoperative treatments for patellofemoral pain syndrome (PFPS). BACKGROUND: Systematic reviews of the quality and usefulness of clinical trials allow for efficient synthesis and dissemination of the literature, which should facilitate clinicians' efforts to incorporate principles of evidence-based practice in the clinical decision-making process. METHODS AND MEASURES: Using a scale based on criteria in the Cochrane Collaboration Handbook, we sought to critically appraise the methodology used in RCTs related to the nonoperative management of PFPS, synthesize and interpret our results, and report our findings in a user-friendly fashion. A scale to assess the methodological quality of trials was designed and pilot tested for its content and reliability. Published RCTs identified during a literature search were then selected and rated by 6 raters. We used predefined cutoff scores to identify specific weaknesses in the clinical research process that need to be improved in future clinical trials. RESULTS: The quality scale we developed was demonstrated to be sufficiently reliable to warrant interpretation of the reviewers' findings. The percentage of trials that met a minimum level of quality for each specific criterion ranged from a low of 25% for the adequacy of the description of the randomization procedure to a high of 95% for the description and standardization of the intervention. CONCLUSIONS: Based on the results of trials exhibiting a sufficient level of quality, treatments that were effective in decreasing pain and improving function in patients with PFPS were acupuncture, quadriceps strengthening, the use of a resistive brace, and the combination of exercises with patellar taping and biofeedback. The use of soft foot orthotics in patients with excessive foot pronation appeared useful in decreasing pain. In addition, at a short-term follow-up, patients who received exercise programs were discharged earlier from physical therapy. Unfortunately, most RCTs reviewed contained qualitative flaws that bring the validity of the results into question, thus diminishing the ability to generalize the results to clinical practice. These flaws were primarily in the areas of randomization procedures, duration of follow-up, control of cointerventions, assurance of blinding, accountability and proper analysis of dropouts, number of subjects, and the relevance of outcomes. Also, given the limited number of high-quality clinical trials, recommendations about supporting or refuting specific treatment approaches may be premature and can only be made with caution. PMID- 12570283 TI - Articular and muscular impairments in cervicogenic headache: a case report. AB - STUDY DESIGN: Case report. OBJECTIVES: To describe the use of manual therapy and muscle re-education for an individual with suspected cervicogenic headache. BACKGROUND: This patient was a 27-year-old woman with complaint of headache. She was functionally limited with prolonged postures and lifting. She also demonstrated impairments in cervical mobility and muscular performance. METHODS AND MEASURES: This patient was treated 8 times over an 8-week period. Intervention included manual upper cervical spine mobilization techniques, muscle re-education for the deep neck flexor muscles, and scapular stabilization exercises. RESULTS: Following treatment, the patient demonstrated an increase in cervical mobility, improved muscular performance, a decrease in headaches, and complete resolution of functional limitations. CONCLUSION: The combination of manual therapy and muscle re-education was successful in relieving headaches and improving function in this patient. PMID- 12570284 TI - Treatment of fabella syndrome with manual therapy: a case report. AB - STUDY DESIGN: Clinical case report. OBJECTIVES: To educate clinicians about fabella syndrome as a possible cause for posterolateral knee pain and dysfunction. Also to describe a physical therapy intervention strategy for posterolateral knee pain secondary to hypomobility or malposition of a fabella. BACKGROUND: A 44-year-old, physically fit, Caucasian male with a 10-year history of left posterolateral knee pain and functional limitations during athletic activities, walking, and activities of daily living presented for evaluation and treatment. He had previously experienced relief of symptoms after experimenting with a mechanical maneuver administered by his wife. METHODS AND MEASURES: A thorough examination for strength, range of motion, and accessory motions was performed. A fabella was palpable in the lateral head of the gastrocnemius muscle and a provisional diagnosis of fabella syndrome was made. While in a prone position, the patient received soft tissue mobilization of the lateral gastrocnemius, followed by medial, lateral, and inferior glides of the fabella. RESULTS: The patient reported an immediate reduction in posterolateral knee pain and demonstrated a 30 degrees increase in active knee flexion. CONCLUSIONS: Physical therapists may be unaware that fabella syndrome is a possible source of posterolateral knee pain and dysfunction. This simple manual therapy intervention was effective in reducing symptoms of pain and increasing tolerance for activities involving knee flexion, extension, and rotation. Physical therapists may wish to add this diagnosis and the corresponding examination and intervention techniques to their management strategy for patients with fabella syndrome. PMID- 12570285 TI - Improvement in sit-up performance associated with 2 different training regimens. AB - STUDY DESIGN: Factorial experimental design. OBJECTIVE: To compare the outcomes of 2 different abdominal muscular fitness training regimens on sit-up performance across sex and abdominal muscular fitness level. BACKGROUND: Researchers suggest that the curl-up, when compared to the sit-up, optimizes the challenge to the abdominal muscles while minimizing shear and compressive forces on the lumbar spine. Although researchers have compared curl-ups and sit-ups in many ways, a comparison of sit-up performance after training programs involving curl-ups and sit-ups has not been investigated. METHODS AND MEASURES: One hundred two active, healthy, college-aged subjects participated in this study. After stratification based upon maximal 2-minute sit-up performance during the orientation session, subjects were randomly assigned to either a training group using curl-up exercise, a training group using sit-up exercise, or a control group. Maximal 2 minute sit-up test performance was measured before and after a 6-week training program. Data were analyzed utilizing an ANOVA model. Significant interactions or main effects were analyzed utilizing Tukey's Honestly Significant Difference Test. Level of significance for all testing was at alpha = 0.05. RESULTS: The sit up training group improved significantly (P < 0.05). No significant difference in sit-up performance was noted for the curl-up or control groups after the 6-week training program. No statistically significant difference in improvement was noted between sex of subject and level of abdominal muscular fitness of subject. CONCLUSIONS: Short-term sit-up training with the Modified Kersey Method in this population significantly improved a maximum 2-minute sit-up test performance. Curl-up training utilizing the same method did not result in improvement in the number of sit-ups performed in 2 minutes. Specificity of training provides the primary explanation for our findings. PMID- 12570286 TI - Bilateral rearfoot asymmetry and anterior knee pain syndrome. AB - STUDY DESIGN: Single-session observational study. OBJECTIVES: To examine the magnitude and extent of asymmetry between right and left rearfoot angles in individuals who are asymptomatic versus symptomatic for anterior knee pain. BACKGROUND: Asymmetry in lower limb measures has been linked to the occurrence of knee pathologies, yet the practice of deriving unilateral rather than bilateral data and thereafter averaging the findings may play a role in obscuring the true nature of the rearfoot angle-knee pain relationship. METHODS AND MEASURES: A convenience sample of participants was recruited from the local university and business communities and a university-affiliated physiotherapy clinic. Eighty-on young adult males and females volunteered to participate in this study. Following an extensive screening process, including mandatory exclusion for a previous history of lower limb dysfunction or injury, data were collected from 75 individuals classified as asymptomatic (n = 50), unilaterally symptomatic (n = 11), or bilaterally symptomatic (n = 14) for anterior knee pain. Right and left rearfoot angles were goniometrically measured under static, weight-bearing conditions. RESULTS: No significant differences in mean right and left rearfoot angles by group or limb were observed. However, these mean values did not always reflect the true variation of data within the sample. Fifty-six percent of the individuals studied demonstrated a minimum of 4 degrees bilateral rearfoot angle difference, while for 20% of the sample this difference exceeded 7 degrees. Only a we yet statistically significant correlation existed between right and left rearfoot angles (r = 0.31, P < 0.01). CONCLUSIONS: Rearfoot angles in weight bearing are not bilaterally symmetric, and the magnitude and direction of the observed asymmetry does not appear to be indicative of whether an individual is asymptomatic or symptomatic for anterior knee pain. PMID- 12570287 TI - Matrix method to find a new set of Zernike coefficients from an original set when the aperture radius is changed. AB - A matrix method is developed that allows a new set of Zernike coefficients that describe a surface or wave front appropriate for a new aperture size to be found from an original set of Zernike coefficients that describe the same surface or wave front but use a different aperture size. The new set of coefficients, arranged as elements of a vector, is formed by multiplying the original set of coefficients, also arranged as elements of a vector, by a conversion matrix formed from powers of the ratio of the new to the original aperture and elements of a matrix that forms the weighting coefficients of the radial Zernike polynomial functions. In developing the method, a new matrix method for expressing Zernike polynomial functions is introduced and used. An algorithm is given for creating the conversion matrix along with computer code to implement the algorithm. PMID- 12570288 TI - Cramer-Rao sensitivity limits for astronomical instruments: implications for interferometer design. AB - Multiple-telescope interferometry for high-angular-resolution astronomical imaging in the optical-IR-far-IR bands is currently a topic of great scientific interest. The fundamentals that govern the sensitivity of direct-detection instruments and interferometers are reviewed, and the rigorous sensitivity limits imposed by the Cramer-Rao theorem are discussed. Numerical calculations of the Cramer-Rao limit are carried out for a simple example, and the results are used to support the argument that interferometers that have more compact instantaneous beam patterns are more sensitive, since they extract more spatial information from each detected photon. This argument favors arrays with a larger number of telescopes, and it favors all-on-one beam-combining methods as compared with pairwise combination. PMID- 12570289 TI - Focal shift in small-Fresnel-number focusing systems of different relative aperture. AB - Axial irradiance distribution arising from the diffraction of a uniform, converging, spherical wave at a circular aperture is studied on the basis of scalar boundary-diffraction wave theory. The combined effects of Fresnel number and angular aperture on the focal shift are evaluated, and the validity of the results is checked against the Kirchhoff boundary conditions. PMID- 12570290 TI - Least-squares phase estimation with multiple parameters in phase-shifting electronic speckle pattern interferometry. AB - We have developed an accurate and robust phase-estimation method in phase shifting electronic speckle pattern interferometry. Unlike other methods that assume a constant phase within a fitting window, our method treats the phase variation with a gradient. A cost function that can utilize the information of pixel positions is formulated on the basis of a least-squares criterion. Powell's iteration method is applied to it to derive the phase and its gradient. An automatic consistency-checking routine and an algorithm that improves the initial guess of the iteration are developed for severe situations with large noise and steep phase variations. PMID- 12570291 TI - Dot pattern generation technique using molecular dynamics. AB - We have developed a new technique for generating homogeneously distributed irregular dot patterns useful for optical devices and digital halftoning technologies. To introduce irregularity, we use elaborately designed sequences called low-discrepancy sequences instead of pseudorandom numbers. We also use a molecular-dynamics redistribution method to improve the distribution of dots. Our method can produce arbitrary density distributions in accordance with a given design. The generated patterns are free from visible roughness as well as any moire patterns when superimposed on other regular patterns. We demonstrate that our method effectively improves luminance uniformity and eliminates moire patterns when used for a backlight unit of a liquid-crystal display. PMID- 12570292 TI - Rigorous model of the scattering of a focused spot by a grating and its application in optical recording. AB - We describe a rigorous model for the scattering of a three-dimensional focused spot by a one-dimensional periodic grating. The incident field is decomposed into a sum of quasi-periodic fields, and the scattering of each of these is computed inside one unit cell of the grating. The model is applied to the simulation of the readout of a DVD disk. The polarization dependence of the reflected near and far fields is studied, and, for a TM-polarized incident spot, plasmons are observed in the reflected far-field intensity. PMID- 12570293 TI - Equivalent circuit of Bragg gratings and its application to Fabry-Perot cavities. AB - An equivalent circuit of both uniform and apodized symmetrical Bragg gratings is proposed, consisting of an ideal partially reflecting mirror placed between two wavelength-dependent uniform propagating sections. The model is simple, exact, and valid for every wavelength; it can be combined with other port-based models and is of great aid in the analysis and design of devices containing two or more Bragg gratings. As an application, the synthesis of dispersive Bragg gratings based Fabry-Perot cavities is demonstrated. The conditions under which a Bragg grating-based Fabry-Perot cavity behaves exactly as an ideal dispersive Fabry Perot cavity are discussed. Experimental results that confirm the theoretical spectral response calculated by using the equivalent circuit are reported. PMID- 12570294 TI - Design of space-variant diffractive polarization elements. AB - Improved diffraction efficiencies can be obtained in the paraxial domain of diffractive optics by considering light explicitly as an electromagnetic rather than a scalar field because of the extra freedoms provided by the state of polarization. For example, diffractive beam splitters with 100% efficiency are made possible by means of space-variant subwavelength-carrier surface-relief elements. Some aspects of the general design theory of polarization-modulating elements for vector fields, including design freedoms and constraints, are presented. Upper bounds of diffraction efficiency are derived and compared with those for the scalar case. Iterative design algorithms are developed. Several design examples with different constraints are presented, and the effects of replacing continuous-fringe structures by pixel structures containing locally linear gratings are evaluated. PMID- 12570295 TI - Unambiguous coherence retrieval from intensity measurements. AB - It is demonstrated that the members of different classes of two-dimensional fields that have the same intensity distributions everywhere in free space but different coherence properties can be identified by measuring the fields' intensities after passing the fields through an anamorphic optical system, such as a cylindrical lens. In this way the ambiguity in coherence determination from intensity measurements alone, present for the case of two-dimensional fields, is removed. PMID- 12570296 TI - Coherent reflectance in a system of random Mie scatterers and its relation to the effective-medium approach. AB - We consider the coherent reflection and transmission of electromagnetic waves from a slab of a dilute system of randomly located, polarizable, spherical particles. We focus our attention on the case where the size of the spheres is comparable to the wavelength of the incident radiation. First, using wave scattering and Mie theories, we derive expressions for the coherent fields that are transmitted and reflected by a very thin slab. Then we find the effective current distribution that would act as a source of these fields. We conclude that if the effective currents were induced in an effective medium, this medium must possess, besides an effective electric permittivity, also an effective magnetic permeability. We find that both of these optical coefficients become functions of the angle of incidence and the polarization of the incident wave. Then we calculate the reflection coefficient of a half-space by considering a semi infinite pile of thin slabs and compare the result with Fresnel relations. Numerical results are presented for the optical coefficients as well as for the half-space reflectance as a function of several parameters. The reflectance is compared with that obtained without considering the magnetic response. Finally, we discuss the relevance and the physics behind our results and indicate as well the measurements that could be performed to obtain an experimental verification of our theory. PMID- 12570297 TI - Feasibility of calculating morphology-dependent resonance peaks by numerical methods. AB - We investigate the feasibility of numerically calculating morphology-dependent resonance (MDR) peaks. To do so, one has to calculate the scattering intensities numerically and determine how difficult it is to numerically predict the position and the magnitude of the MDR peaks. However, at present, in practice it is impossible to calculate MDR peaks with a personal computer because so much computing time is required. Therefore the surface values of the Debye potential and its derivative for a homogeneous sphere are obtained from Mie's analytical solution and then used in integral equations to give the scattering intensities at a specific position of infinity by numerical integrations. It is shown that if a sufficient number of surface elements are used, the MDR peaks are exactly calculated for a homogeneous sphere with refractive index of 1.5, 1.4, and 1.3 up to a size parameter of 20. One can conjecture the number of finite and boundary elements necessary to numerically compute accurate scattering intensities. It should be also noted that the number of surface elements necessary for exact integration shows peaks similar to MDR peaks with respect to the size parameter. Therefore one will need many more elements at the size parameter at which the MDR occurs. PMID- 12570298 TI - Resonant inelastic scattering by use of geometrical optics. AB - We investigate the inelastic scattering on spherical particles that contain one concentric inclusion in the case of input and output resonances, using a geometrical optics method. The excitation of resonances is included in geometrical optics by use of the concept of tunneled rays. To get a quantitative description of optical tunneling on spherical surfaces, we derive appropriate Fresnel-type reflection and transmission coefficients for the tunneled rays. We calculate the inelastic scattering cross section in the case of input and output resonances and investigate the influence of the distribution of the active material in the particle as well as the influence of the inclusion on inelastic scattering. PMID- 12570299 TI - Adaptive phase-shifting algorithm for temporal phase evaluation. AB - Most standard temporal-phase-shifting (TPS) algorithms evaluate the phase by computing a windowed Fourier transform (WFT) of the intensity signal at the carrier frequency of the system. However, displacement of the specimen during image acquisition may cause the peak of the transform to shift away from the carrier frequency, leading to phase errors and even unwrapping failure. We present a novel TPS method that searches for the peak of the WFT and evaluates the phase at that frequency instead of at the carrier frequency. The performance of this method is compared with that of standard algorithms by using numerical simulations. Experimental results from high-speed speckle interferometry studies of carbon fiber panels are also presented. PMID- 12570300 TI - Delay and dispersion characteristics of a frequency-domain optical delay line for scanning interferometry. AB - The reflective frequency-domain optical delay line employing a diffraction grating, a lens, and a tiltable mirror has emerged as a device particularly suitable for interferometry and optical coherence tomography. The device is comprehensively described, both theoretically and experimentally, in the context of interferometry. The variations of phase and group delay produced by the device as well as its dispersive properties are described and demonstrated experimentally. PMID- 12570301 TI - Polarization state fixer composed of passive optical devices. AB - We investigate a polarization state fixer that produces light with an arbitrary desired polarization state and constant power from an arbitrary elliptically polarized input light whose principal polarization axis fluctuates randomly. An example of such a device composed of a quarter-wave plate and a polarizer is proposed, and its characteristics at wavelengths of 633 nm and 1550 nm are demonstrated. We conclude that we cannot obtain a passive device that produces fixed-axis, linearly polarized light with a constant power level from an elliptically polarized light with a varying elliptical parameter. PMID- 12570302 TI - Generalized far-infrared magneto-optic ellipsometry for semiconductor layer structures: determination of free-carrier effective-mass, mobility, and concentration parameters in n-type GaAs. AB - We report for the first time on the application of generalized ellipsometry at far-infrared wavelengths (wave numbers from 150 cm(-1) to 600 cm(-1)) for measurement of the anisotropic dielectric response of doped polar semiconductors in layered structures within an external magnetic field. Upon determination of normalized Mueller matrix elements and subsequent derivation of the normalized complex Jones reflection matrix r of an n-type doped GaAs substrate covered by a highly resistive GaAs layer, the spectral dependence of the room-temperature magneto-optic dielectric function tensor of n-type GaAs with free-electron concentration of 1.6 x 10(18) cm(-3) at the magnetic field strength of 2.3 T is obtained on a wavelength-by-wavelength basis. These data are in excellent agreement with values predicted by the Drude model. From the magneto-optic generalized ellipsometry measurements of the layered structure, the free-carrier concentration, their optical mobility, the effective-mass parameters, and the sign of the charge carriers can be determined independently, which will be demonstrated. We propose magneto-optic generalized ellipsometry as a novel approach for exploration of free-carrier parameters in complex organic or inorganic semiconducting material heterostructures, regardless of the anisotropic properties of the individual constituents. PMID- 12570303 TI - Cartilage thickness measurements from optical coherence tomography. AB - We describe a new semiautomatic image processing method for detecting the cartilage boundaries in optical coherence tomography (OCT). In particular, we focus on rabbit cartilage since this is an important animal model for testing both chondroprotective agents and cartilage repair techniques. The novel boundary detection system presented here consists of (1) an adaptive filtering technique for image enhancement and speckle reduction, (2) edge detection, and (3) edge linking by graph searching. The procedure requires several steps and can be automated. The quantitative measurements of cartilage thickness on OCT images correlated well with measurements from histology. PMID- 12570304 TI - Theoretical investigation of the signal-to-noise ratio in fluorescence lifetime imaging. AB - We deduce the signal-to-noise ratio for fluorescence lifetime imaging when using frequency-domain methods. We assume mono-exponential decay and quantum-noise limited performance. The results are compared with Monte Carlo simulations with good agreement. We also compare our results with previous investigations of time domain methods for fluorescence lifetime imaging. For a given number of detected photons, we find that frequency-domain and time-domain methods are equally good. The correct choice of detection technique and its parameters is important for obtaining good results. PMID- 12570305 TI - Optical sectioning by holographic coherence imaging: a generalized analysis. AB - A theory of optical sectioning by image plane holography is developed, emphasizing the use of broad-spectrum holographic methods to enhance the process. It is shown that a broad-spectrum source in a grating interferometer imitates the behavior of a monochromatic broad source. PMID- 12570306 TI - Spatial width and power-content ratio of hard-edge diffracted beams. AB - On the basis of the intensity-moment formalism, certain analytical relationships are obtained for both the angular domain and the size of a transverse region of the beam that ensure a power content of at least 75% of the total power. As an illustrative application, the analytical results are compared with the exact values (numerically computed) of the amplitude of a lowest-order Gaussian beam diffracted by slits. PMID- 12570307 TI - EC adopts revised legislation on FMD. PMID- 12570308 TI - Effect of month of birth on hip dysplasia in labrador retrievers and Gordon setters. AB - Recent studies have proposed selective breeding policies for preventing or controlling hip dysplasia, based upon the relationships between the hip scores of the offspring and their parents. However, these studies have ignored the possible effects on an animal's hip score of its age when it was examined and the month in which it was born. This paper describes the results of statistical analyses of large data sets of Kennel Club-registered labrador retrievers and Gordon setters. Regression modelling showed that an animal's hip score depended significantly upon the hip score of its sire and dam, and upon its month of birth. Enlarging the model by including its age when examined made it possible to quantify the significant genetic parental effects and the effects of the animal's month of birth and its age when examined. PMID- 12570309 TI - Transmission of porcine reproductive and respiratory syndrome virus by houseflies (Musca domestica). AB - Three hundred houseflies were allowed to feed on donor pigs viraemic with porcine reproductive and respiratory syndrome virus (PRRSV) on the fifth, sixth and seventh days after the pigs had been inoculated with the virus. After 60 seconds, the flies' feeding was interrupted, and they were transferred manually to feed to repletion on a naive recipient pig housed in a separate room. To enhance the chance of the flies obtaining the pigs' blood, the back of each pig was scarified with sandpaper until a slight haemorrhage was visible. The PRRSV was transmitted from the donor to the recipient pigs, and PRRSV RNA was detected by reverse transcriptase-PCR from homogenates of the flies. In a second experiment, 210 houseflies were allowed to feed to repletion on a PRRSV-infected pig on the sixth day after it had been inoculated, and were then maintained under laboratory conditions. Groups of 30 flies were collected immediately after they had fed and six, 12, 24, 48, 72 and 96 hours later, and were tested for PRRSV. Homogenates of the flies collected up to six hours after feeding were PCR- and pig bioassay positive, but the others were negative by both tests. PMID- 12570310 TI - Study of dog and cat owners' perceptions of medical treatment for cancer. AB - Between April 10 and June 9, 2000, 91 owners of cats and dogs that were being or had been treated with anticancer chemotherapy were given a questionnaire designed to obtain information about their experiences as a result of the discovery and treatment of the disease, the efficacy and side effects of the treatment and their perceptions of the procedures associated with the administration of the drugs. Nearly all of the owners felt that the treatment was worthwhile. The level of observed side effects was low. Well over half of the owners believed that their animal had lived longer than it would have if it had not been treated and that its general wellbeing had improved. In general, they felt that the treatment had been rewarding and that any adverse side effects had been outweighed by the positive experiences during the treatment; they felt that they had been well informed and that their animals had benefited from the treatment. PMID- 12570311 TI - Reduced antibiotic resistance to fluoroquinolones and streptomycin in 'animal friendly' pig fattening farms in Switzerland. PMID- 12570312 TI - Faecal culture of wild animals for Escherichia coli O157:H7. PMID- 12570313 TI - Copper-induced hepatitis in an Anatolian shepherd dog. PMID- 12570314 TI - Bovine ephemeral fever in beef cattle herds in the Jordan Valley, Israel. PMID- 12570315 TI - Ciprofloxacin-resistant Campylobacter. PMID- 12570317 TI - Vets in art. PMID- 12570316 TI - Uterine prolapse and mid-pregnancy uterine torsion in cows. PMID- 12570318 TI - A foal from a mule in Morocco. PMID- 12570319 TI - Norovirus activity--United States, 2002. AB - During June-December 2002, an increased number of outbreaks of acute gastroenteritis (AGE) were reported on cruise ships sailing into U.S. ports. In addition, since October 2002, several states have noted an increase in outbreaks of AGE consistent clinically and epidemiologically with norovirus infection, particularly in institutional settings such as nursing homes (CDC, unpublished data, 2002). This report describes recent norovirus activity in two states and New York City (NYC) and data from CDC that indicate the possible emergence of a predominant, circulating norovirus strain. PMID- 12570320 TI - Deaths among drivers of off-road vehicles after collisions with trail gates--New Hampshire, 1997-2002. AB - During April-July 2002, three deaths occurred on New Hampshire trails when adolescents driving off-highway recreational vehicles (OHRVs) collided with trail gates. Because of these three incidents, the New Hampshire Department of Health and Human Services conducted a study to determine the extent of the problem and characteristics of the fatal events. This report describes trail gate fatalities in New Hampshire during 1997-2002. To prevent trail gate collisions, efforts should focus on increased enforcement of OHRV operating rules, driver education, enhanced gate visibility, and improved signage. PMID- 12570321 TI - Human rabies--Iowa, 2002. AB - On September 28, 2002, a man aged 20 years residing in Linn County, Iowa, died from rabies encephalitis caused by infection with a variant of rabies virus associated with silver-haired (Lasionycteris noctivagans) and eastern pipistrelle (Pipistrellus subflavus) bats. This is the first case of human rabies in Iowa since 1951. This report summarizes the investigation of the case by the Linn County and Iowa public health departments. Bats found in living quarters should be submitted to local public health laboratories for rabies testing. PMID- 12570322 TI - Combined opto-electronic perometry and bioimpedance to measure objectively the effectiveness of a new treatment intervention for chronic secondary leg lymphedema. AB - Secondary lymphedema of the legs is a common sequela of patients treated for cancer of the reproductive, gastrointestinal, urinary systems and melanoma. From a clinical and research perspective it is of utmost importance to use techniques that objectively quantify leg volume and fluid composition as an indicator of lymphedema severity and response to treatment. Two techniques often used in both the clinical and research setting are leg perometry and multi-frequency bioimpedance. Although both techniques have been extensively validated, this trial aimed to cross correlate both measurement techniques to ascertain whether each or both could be used reliably for measurement of leg lymphedema. These measurements were utilized throughout a clinical trial that assessed the effectiveness of a new home based treatment program in the form of the Sun Ancon Aerobic Exerciser. This machine delivered both elevation and passive exercise to the legs, with participants using the machine over a three week period during which time their leg volumes were measured using both perometry and bioimpedance. The results demonstrated that leg volume measurements decreased using both perometry and bioimpedance. The reduction in body extracellular fluid as measured by bioimpedance correlated well with a reduction in leg volume as measured by perometry. Bioimpedance also recorded a reduction in weight, which was correlated with the reduction in leg volume as measured by perometry. This trial confirms that perometry and bioimpedance were both effective in independently showing a reduction in leg lymphedema using the Aerobic Exerciser therapy, and that both methods can be reliably used to measure and follow leg lymphedema. PMID- 12570323 TI - Effect of phosphodiesterase III inhibitor (Olprinone) on thoracic duct lymph flow in anesthetized sheep with experimentally induced heart failure by endothelin-1. AB - We investigated the short-term effects of a phosphodiesterase III inhibitor (Olprinone) on hemodynamics and thoracic duct lymph flow in anesthetized open chest sheep with heart failure induced by endothelin-1 (cardiogenic shock). Ultrasound transit-time flow probes were attached to the thoracic duct, the ascending aorta and the renal artery. Arterial, pulmonary and central venous pressures were monitored. Endothelin-1 was infused intravenously at a dosage that reduced cardiac output to 50% or more of baseline (n=11). The effects of Olprinone were examined (n=5) by intravenous infusion after endothelin-1 administration. Other sheep (n=6) were used as controls. Olprinone significantly increased cardiac output that had been decreased by endothelin-1 and further increased thoracic duct flow that had been increased by endothelin-1. Increased arterial and pulmonary pressures induced by endothelin-1 administration were rapidly decreased by Olprinone. Renal arterial flow and central venous pressure were, however, unchanged by Olprinone. Overall, Olprinone acutely improved experimental cardiogenic shock (heart failure) induced by endothelin-1, and maintained thoracic duct lymph flow at a high level after endothelin-1 administration. PMID- 12570324 TI - Lymphatic pathways of the upper medial quadrant of the breast in healthy women: radiotracer study of the sentinel lymph node. AB - This study examined the lymphatic drainage after injection of a radiotracer into the upper medial quadrant of the breast in healthy women. Most studies of lymphatic pathways of the breast have been performed in patients with breast cancer and concentrate on the upper lateral quadrant of the breast because of the high incidence of carcinoma at this site. The lymphatic drainage pathways of the medial half of the breast, however, has been less studied. A radiotracer (Tc-99m human serum albumin nanocolloid or HSA) was injected intradermally into the upper medial quadrant of the right breast in 12 healthy women. Dermal markers were placed at the middle of the clavicle, the axilla and at the jugular incisura. Three minutes after injection a static image of the injection site was made with a scintillation camera (Multispect 2 Gamma Camera System) over 20 seconds. After nine minutes, local soft massage was instituted at the injection site for 6 minutes. Fifteen minutes after injection, a graphic scintigraphic image was made of both breasts and axillae over 22 minutes. After this interval, three or four static images were made for a few seconds to locate the sentinel lymph node as related to the injection site. A sentinel lymph node (lymphatic pathway) in the axilla was visualized in 11 subjects (91.9%) and was undetected in one subject (8.3%). The radiotracer migrated in all patients (100% ) towards the ipsilateral axilla. In 9 subjects, the sentinel lymph node was visualized 15 minutes after injection, whereas in 2 subjects it appeared within an hour. PMID- 12570325 TI - Axillary lymph drainage as a prognostic factor of survival in breast cancer. AB - Axillary lymph node staging is an important prognostic factor in patients with breast cancer. The long-term survival is poor if the number of metastatic axillary lymph nodes exceeds three. With 1-3 metastatic lymph nodes, survival in 30% of the patients is 20 years. The markedly different outcome of these patients with metastatic axillary lymph nodes suggests that other factors likely play a critical role in breast cancer dissemination. In this study, we examined the association of impaired axillary lymph drainage with breast cancer survival. Fifty patients with breast cancer underwent direct breast lymphography before treatment to evaluate the status of ipsilateral axillary lymph nodes and lymph drainage at time of presentation. Twenty-five patients with signs of metastatic disease and/or impaired lymph drainage were followed for 10 years. Seven of 25 patients survived 10 years; 6 are tumor free; and 1 has distant mestastases. The other 18 patients died during the 10 year period. Seventeen of the 18 died from disseminated breast cancer; 1 died of unrelated disease. In the survivors, direct breast lymphography showed normal breast lymph drainage. All patients with distant metastases had obstructed lymph vessels at the time of original diagnosis. These findings suggest that the chance for survival is determined both by the status of axillary lymph drainage as well as the number of metastatic axillary lymph nodes. PMID- 12570327 TI - Lymph heart scintigraphy in the toad. AB - To our knowledge, there are no reports on the use of radionuclide labeled agents to image the lymphatic system of anuran amphibia. Lymphoscintigraphy was performed in 18 toads Bufo marinus L, after the injection of Tc-99m-serum albumin within the ventral lymph sac. Images were obtained with a rectilinear scanner immediately after tracer administration. Scintigrams were digitalized and densitometric profiles were obtained. One or more lymph hearts were demonstrated on all 18 scans. Scintigrams showed the four lymph hearts functioning simultaneously in only 17% of the toads. Lymphoscintigraphy with Tc-99m serum albumin is useful to study the movement of lymph from the lymph sacs or to study the function of lymph hearts in amphibia. PMID- 12570326 TI - The effects of Unguentum Lymphaticum on skin in patients with obstructive lymphedema of the lower extremities. AB - Obstructive lymphedema of extremities in humans is characterized by swelling of tissues with lymph stasis and inflammatory infiltrates in skin and subcutaneous tissues. Treatment of the inflammatory component requires application of antiinflammatory drugs. We studied the effect of topical application of Unguentum Lymphaticum (UL) containing antiphlogistic compounds (digitalis, calendulin, hyoscyamine, colchicine and podophyllin) on lymphedematous skin in 33 patients with stage II postinflammatory obstructive lymphedema. A three-week treatment of swollen legs with UL brought about stimulation of epidermal cells with proliferation of keratinocytes, increased numbers of Langerhans cells, accumulation of macrophages in the dermis and activation of infiltrating cells and endothelia. Besides some foci of acanthosis, no degenerative changes were observed in the skin in patients treated for 12 weeks and no changes were observed in the placebo treated groups. Immunohistochemical evaluation of biopsy specimens of inguinal lymph nodes in patients treated for 12 weeks revealed reactive granulocyte and macrophage accumulation in the cortical and paracortical areas. Components of UL inhibited stimulation of blood mononuclear cells in in vitro cultures. UL did not change the spectrum of calf skin bacterial populations. The nonspecific stimulation of skin-associated lymphoid tissue and putative elimination of factors responsible for lymphe-dematous skin inflammation appears to be responsible for the beneficial clinical effect of UL on legs with lymph stasis. PMID- 12570328 TI - Lifestyle, health and disease prevention: the underlying mechanisms. AB - International studies in geographic pathology provide background information that a disease may have a quite different incidence and resulting mortality as a function of area of residence. Investigations in animals can model fairly precisely what is learned through such international research, and provide the basis for examining relevant hypotheses and, more importantly, possible mechanisms of action. These approaches can yield public health recommendations and health promotion activities. Regular intake of foods rich in saturated fats, such as meat and certain dairy products, raises the risk of coronary heart disease, especially in smokers. The total mixed fat intake is associated with a higher incidence of the nutritionally linked cancers (i.e. of the postmenopausal breast, distal colon, prostate, pancreas, ovary and endometrium). Monounsaturated oils, such as olive or canola oil, are low-risk fats, as shown in animal models, and through the finding that the incidence of coronary heart and neoplastic diseases is lower in the Mediterranean region, where such oils are customarily used. Fish and fish oils are protective. The associated genotoxic carcinogens for several of these cancers, and also in heart disease causation, are heterocyclic amines, produced during the broiling and frying of creatinine-containing foods such as meats. Excessive salt intake is associated with high blood pressure and with stomach cancer, especially with inadequate intake of potassium, from fruits and vegetables, and calcium from certain vegetables and low-fat dairy products. Bran cereal fiber intake, especially with adequate calcium, yields an increased stool bulk, eliminating factors involved in colon and breast cancer. Vegetables and fruits, as well as soy products, are rich in antioxidants that are essential to lower disease risk stemming from reactive oxygen species in the body. Green and black tea are excellent sources of such beneficial antioxidants of a polyphenol nature, as are cocoa and chocolates. Antioxidants also extend healthy aging and may protect against Alzheimer's and Parkinson's diseases. Nutritional lifestyles can be described for most populations in the world and offer the possibility of a healthy long life. PMID- 12570329 TI - Plant antimutagens and their mixtures in inhibition of genotoxic effects of xenobiotics and aging processes. AB - The antimutagenic effect of the bioactive compounds from fruits of Morus alba L. (MA), Punica granatum L. (PG), Diospyros kaki L. (DK), Cydonia oblonga Mill. (CO) and roots of Glycyrrhiza glabra (GG) were investigated. The antimutagenic effects of compounds separately as well as their action in complex mixtures were studied on mutations induced by genotoxicants (X-rays, N-methylnitrosourea, cyclophosphamide, NaF) and aging in bone marrow cell chromosomes from mice and rats. When tested separately and in a complex mixture, the plant products showed an ability to decrease the frequency of chromosome aberrations. The antimutagenic properties of the complex mixtures were considerably greater than those of the separate components. More antimutagenic activity of the mixture was revealed when mutagenesis was the result of X-rays and the natural aging processes. PMID- 12570330 TI - Environment-gene interactions in intestinal cancer. AB - Only 5-10% of all colorectal cancer in the United States can be directly attributed to inheritance of genetic predisposition for tumor development. Thus, the vast majority of colorectal cancer is classified as sporadic, and in these patients environmental factors--particularly the diet--play a major role in determining the probability of tumor formation and its progression. Investigations of how dietary components interact with genetic factors in cancer development have been extremely productive in terms of understanding the subtle and complex mechanisms that maintain homeostasis of the intestinal mucosa, and how perturbations in these mechanisms cause disease. We have found that the cyclin-dependent kinase inhibitor p21(WAF1/cip1) plays a major role in regulating several aspects of mucosal homeostasis and the response to dietary and pharmacologic modulators of tumorigenesis; that disruption of lineages of differentiation of intestinal epithelial cells are intimately involved in tumor formation; and that important pathways that contribute to normal homeostasis and cancer development may be coordinately regulated by mitochondrial function, with the mitochondrial membrane potential playing a key role. Several lines of evidence from our work have also suggested that intestinal epithelial cells have adapted to the environment that they usually encounter. This renders the cells competent to efficiently utilize factors in the intestinal lumen in normal metabolic and signaling pathways that contribute to homeostasis. PMID- 12570331 TI - Chemoprevention strategies for prostate cancer. AB - Prostate cancer is the most common male malignancy in western countries. Although primary prevention of prostate cancer is not possible, screening using prostate specific antigen (PSA) may eliminate prostate cancers by definitive treatments. Prevention of clinically detectable prostate cancer requires earlier chemoprevention interventions. Because prostate cancer is histologically present in 30-50% of 30- to 50-year-old men, effective chemoprevention needs to inhibit not only prostate carcinogenesis but also growth and progression of these cancers. A prostate carcinogenesis animal model has been used to screen chemopreventive agents; inhibitory effects were found with 9-cis-retinoic acid, dehydroepiandrosterone, fluasterone, and the Bowman-Birk inhibitor and an isoflavone mixture which both occur in soy. Such results can be used to select agents for clinical trials. Besides large-scale long-duration prevention trials, trials of short/intermediate duration using smaller cohorts prior to or following radical prostatectomy may provide excellent and cost-effective approaches for chemopreventive agent efficacy testing. Intervention prior to surgery allows measurements of intervention agents and intermediate end-points in the prostate. These peri-surgical trials only assess inhibition of growth and progression of preexisting cancer, not real preventive effects, but they focus on clinically significant cancers. Such trials are an essential step in the development of antiprostate cancer chemoprevention agents. PMID- 12570332 TI - Inhibitory effects of tea and caffeine on UV-induced carcinogenesis: relationship to enhanced apoptosis and decreased tissue fat. AB - Oral administration of green tea or caffeine to hairless SKH-1 mice for 2 weeks stimulated UV-induced increases in apoptotic sunburn cells in the epidermis, and a similar effect was observed when caffeine was applied topically immediately after UV. In mice pretreated with UV for 22 weeks (high-risk mice without tumors), topical applications of caffeine 5 days a week for 18 weeks with no further UV treatment inhibited carcinogenesis and stimulated apoptosis in the tumors. Oral administration of green or black tea to UV-pretreated high-risk mice for 23 weeks inhibited skin tumorigenesis, decreased the size of the parametrial fat pads and decreased the thickness of the dermal fat layer away from tumors and directly under tumors. Administration of the decaffeinated teas had little or no effect on these parameters and adding caffeine to the decaffeinated teas restored their inhibitory effects. Administration of caffeine alone also inhibited carcinogenesis and decreased the size of the parametrial fat pads and the thickness of the dermal fat layer. Using data from individual mice and linear regression analysis, we found a highly significant positive correlation between the thickness of the dermal fat layer away from tumors and the number of tumors per mouse. PMID- 12570334 TI - An evidence-based approach to cancer prevention clinical trials. AB - Research on carcinogenesis and its inhibition has made significant progress in the last 30 years, providing an impressive body of evidence that supports various strategies for cancer prevention. Innovative studies have helped to identify potential causes of cancer, including environmental factors such as diet, and provided valuable information about their mechanisms of action. Hundreds of epidemiologic and experimental studies have focused on possible associations between dietary factors and different types of cancer. During the same period, potential inhibitors of cancer that appeared able to prevent, arrest or reverse cancer development by interfering with one or more steps in the process of carcinogenesis were identified, and the term 'chemoprevention' was coined for this pharmacological approach to cancer prevention. Promising compounds were systematically evaluated for their potential as chemopreventive agents. Numerous agents were determined to be safe and effective in preclinical trials, including naturally occurring vitamins, minerals and phytochemicals as well as synthetic compounds. Based on preclinical results, selected agents have been and are now being evaluated in phase I, II and III clinical interventions for various cancers. Development of valid surrogate end point biomarkers for clinical disease that can be modulated by interventions is essential to accelerate progress in cancer prevention clinical trials. PMID- 12570333 TI - Microarray analysis of diet-induced alterations in gene expression in the ACI rat prostate. AB - The natural history of prostate cancer is a multistage process that involves the transition from normal tissue to subclinical cancer, with progression to carcinoma in situ and eventually metastatic disease. Evidence suggests that a high-fat diet plays a critical role in the biology and progression of the disease. ACI rats were maintained for two generations on high beef fat or control diets for 18 months. Affymetrix microarrays were used to analyze the mRNA expression levels in the dorsolateral prostates of rats on the different diets. Approximately 4752 genes and expressed sequence tag (EST) were expressed in the prostates of rats on either diet. Twenty-seven genes were upregulated and 28 genes downregulated in the high beef fat diet. Data analysis indicated that a high beef fat diet affects the expression of genes involved in inflammation, glucose and fatty acid metabolism, androgen metabolism, potential tumor suppression and protein kinase activity, as well as intracellular and extracellular matrix molecules, growth factors and androgen responsive genes. Results from these and future studies will lead to a better understanding of the effect of diet on gene expression in the prostate and facilitate the rational design and assessment of potential dietary programs for prostate cancer prevention. PMID- 12570335 TI - Antioxidant and antitumor effects of hydroxymatairesinol (HM-3000, HMR), a lignan isolated from the knots of spruce. AB - The antioxidant properties of hydroxymatairesinol (HM-3000) were studied in vitro in lipid peroxidation, superoxide and peroxyl radical scavenging, and LDL oxidation models in comparison with the known synthetic antioxidants Trolox (a water-soluble vitamin E derivative), butylated hydroxyanisol (BHA) and butylated hydroxytoluene (BHT). On a molar basis HM-3000 was a more effective antioxidant than Trolox in all assays and more effective than BHT or BHA in lipid peroxidation and superoxide scavenging test. The in vivo antioxidative effect (evaluated as the weight gain of C57BL/6J mice fed an alpha-tocopherol-deficient diet) of HM-3000 (500 mg/kg per day) was comparable to that of DL-alpha tocopherol (766 mg/kg per day). The antitumor activity of HM-3000 was studied in dimethylbenz[a]anthracene (DMBA)-induced rat mammary cancer. HM-3000 had a statistically significant inhibitory effect on tumor growth. Prevention of tumor formation was also evaluated in the Apc(Min) mice model, which develops intestinal polyps spontaneously. HM-3000 was given in diet at 30 mg/kg per day and decreased the formation of polyps and prevented beta-catenin accumulation into the nucleus, the pathophysiological hallmark of polyp formation in this mouse model. In short-term toxicity studies (up to 28 days) HM-3000 was essentially non-toxic when given p.o. to rats and dogs (daily doses up to 2000 and 665 mg/kg, respectively); HM-3000 was shown to be well absorbed (> 50% of the dose) and rapidly eliminated. In human studies HM-3000 has been given in single doses up to 1350 mg to healthy male volunteers without treatment-related adverse events. Rapid absorption from the gastrointestinal tract and partial metabolism to enterolactone in humans was demonstrated. In summary, HM-3000 is a safe, novel enterolactone precursor lignan with antioxidant and antitumor properties. PMID- 12570337 TI - Early development of cancer chemoprevention clinical trials: studies of dietary calcium as a chemopreventive agent for human subjects. AB - Early cancer chemoprevention clinical trials in human subjects had to be carried out with large numbers of subjects studied for long durations, measuring cancer as an end point. However new findings on abnormal epithelial cell growth and development during the multistage evolution of colonic tumors made it possible to carry out chemoprevention clinical trials in several stages, with fewer subjects studied for shorter durations, thus enabling investigators to analyze increasing numbers of chemopreventive agents and nutritional regimens in clinical trials. Supplemental dietary calcium was the first candidate chemopreventive agent studied in this multistage approach in human subjects, as a putative agent for colon cancer prevention. Early- and late-stage intermediate biomarker studies in humans have strongly suggested utility for supplemental dietary calcium to inhibit the development of benign and subsequent malignant colonic neoplasms. Preclinical experimental studies have further demonstrated the ability of increased dietary calcium to inhibit the evolution of colonic tumors when they were induced by targeted mutations, dietary factors, and particularly when given over a long duration of lifespan. PMID- 12570336 TI - Strategies for chemoprevention of liver cancer. AB - Liver cancer is one of the most common cancers worldwide. Infection with hepatitis B virus and exposure to aflatoxins in the diet act synergistically to amplify risk. From a public health perspective, hepatitis virus vaccination programs and efforts to both reduce aflatoxin exposures and to attenuate the toxicological consequences of unavoidable exposures should have major impacts on the incidence of this disease. Experimentally, aflatoxin-induced hepatocarcinogenesis can be inhibited by over a score of different chemopreventive agents with multiple mechanisms of action. One agent, oltipraz, is a potent inducer of phase 2 enzymes involved in the detoxication of carcinogens including aflatoxin. A second agent, chlorophyllin, impedes the bioavailability of carcinogens by forming molecular complexes and enhances their elimination in the fecal stream. This review highlights the findings of recent randomized clinical trials with oltipraz and chlorophyllin conducted in individuals exposed to dietary aflatoxins and at high risk for development of liver cancer. Both chemopreventive agents modulated levels of aflatoxin biomarkers in the study participants in manners consonant with protection. PMID- 12570338 TI - Chemoprevention of large bowel carcinogenesis; the role of control of cell proliferation and significance of beta-catenin-accumulated crypts as a new biomarker. AB - Control of cell proliferation is important for cancer prevention since cell proliferation has essential roles in carcinogenesis in the processes of both initiation and promotion. In large bowel carcinogenesis, carcinogens produce hyperproliferation of cells in the target sites and the cell proliferation persists even after the cessation of carcinogen exposure. Chemopreventive agents principally control the increased cell proliferation when given in the initiation as well as post-initiation phases. Aberrant crypt foci (ACF) which appear soon after carcinogen exposure in large bowel carcinogenesis in rodents have been used as a reliable biomarker for screening of potential chemopreventive agents. Recently, our group demonstrated the presence of probable premalignant lesions with frequent beta-catenin gene mutations and accumulation of the corresponding protein in the colonic epithelium of rats given a large bowel carcinogen. Such early-appearing lesions lack the morphological appearance of ACF. Expression of these beta-catenin-accumulated crypts (BCAC) is markedly suppressed by a chemopreventive cyclooxygenase-2 inhibitor, celecoxib. BCAC are suggested to be more reliable biomarkers than ACF for screening effective chemopreventive agents for colorectal cancer and for investigating the mode of action of the agents. PMID- 12570339 TI - Structural concepts in cancer prevention. AB - The notion of developing cancer preventative strategies is attractive both from a public health and from a health economic viewpoint. However, as currently visualized, this may involve dietary supplementation of publicly available foods or the ingestion of specific supplements for prolonged periods of time. In view of the fact that the outcome of such preventative strategies may as yet not be known, it is essential that the strategy is devoid of risks. Structure-activity relationship (SAR) concepts can be of use in identifying possible health hazards associated with chemoprevention. Overall, SAR can be used (1) to predict the chemopreventative potential of a chemical and to understand its mechanism of action, (2) to evaluate the toxicological liabilities of such agents and (3) to design molecules with enhanced chemopreventative potency and decreased (or abolished) toxicity. While SAR techniques currently available are appropriate to achieve these aims, the primary block to their widespread deployment is lack of sufficient experimental data of acceptable quality to perform SAR modeling. The present report analyzes the current applicability of SAR methods to cancer chemoprevention. PMID- 12570340 TI - Multiple molecular targets of indole-3-carbinol, a chemopreventive anti-estrogen in breast cancer. AB - The mechanism of action of the anti-estrogen indole-3-carbinol (I3C), present in cruciferous vegetables, is being examined in our laboratory with a view to promote the use of this naturally occurring chemopreventive as an alternative to synthetic anti-estrogens in human breast cancer. Our previous results clearly demonstrated that despite its low affinity for the estrogen receptor (ER), I3C abrogated estradiol-mediated cellular and biochemical effects in estradiol responsive cells and tissues. In an earlier report, we identified ER phosphorylation as one of the targets of I3C, and in this communication we describe the consequence of inhibition of ER phosphorylation. Estradiol-induced DNA-binding proteins that bound to several DNA-responsive elements were inhibited by I3C and this effect was not at the level of DNA-protein physical interaction as inclusion of I3C in vitro in the reaction mix did not affect the binding. We analyzed the spectrum of genes induced by estradiol and modulated and/or intercepted by I3C. Our results conclude that although estradiol-mediated functions are affected by I3C, its biochemical targets are multiple and some of these may be modulated by the oligomeric products of I3C. PMID- 12570341 TI - Weight control and physical activity in cancer prevention: international evaluation of the evidence. AB - To evaluate the evidence for the role of weight control and physical activity in cancer prevention and to identify priorities for research and for public health action in relation to the primary prevention of cancer, an international working group of experts was convened in Lyon in February 2001 by the International Agency for Research on Cancer of the World Health Organization. The expert group concluded that limiting weight gain during adult life, thereby avoiding overweight and obesity, reduces the risk of postmenopausal breast cancer and cancers of the colon, endometrium, kidney (renal cell) and esophagus (adenocarcinoma). Limiting weight gain possibly reduces risk of cancer of the thyroid. Weight loss among overweight or obese persons possibly reduces risks of these cancers, but no definite conclusion can be drawn because of the paucity of the epidemiological evidence. The working group also concluded that there was sufficient evidence for the role of physical activity in preventing colon and breast cancers, and limited evidence for the cancers of the prostate and endometrium. Some of these effects were independent of that of the weight control. Taken together, the working group considered that excess body weight and physical inactivity account for approximately a quarter to one-third of cancers of the colon, breast, endometrium, kidney (renal cell) and esophagus (adenocarcinoma). Thus adiposity and physical inactivity appear to be the most important avoidable causes of these cancers. PMID- 12570342 TI - Anticarcinogenicity of monocyclic phenolic compounds. AB - The synthetic monocyclic phenolics (MPs), acetaminophen (APAP), butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) are antimutagenic or anticarcinogenic against a diversity of chemical carcinogens affecting a variety of tissues in experimental animals. In studies in this laboratory of the anticarcinogenicity of MPs, the focus has been on delineating efficacy at low levels of MPs that do not elicit adaptive or toxic responses. To accomplish this, we are studying anticarcinogenicity against the neoplastic initiating activity of lower doses of carcinogens than have previously been studied and which are closer to human environmental exposures. In these studies, we have investigated anticarcinogenicity of BHT against liver cancer in rats induced by either 2 acetylaminofluorene (AAF) or aflatoxin B1 (AFB1) and anticarcinogenicity of APAP against colon cancer induced in rats by 3,2'-dimethyl-4-aminobiphenyl (DMAB). BHA and BHT at 100-125 ppm in the diet inhibited the initiation phase of AAF and AFB1 hepatocarcinogenesis and therefore may act intracellularly to block effects of the carcinogen. Likewise, with APAP in colon anticarcinogenicity, at 1000 ppm it reduced DNA binding and exerted a cytoprotective effect against DMAB. Thus, APAP also shows evidence of producing a blocking effect. We conclude that these MPs appear to be anticarcinogenic through a mechanism different from that of most other chemopreventive agents, possibly involving interception of the reactive chemical species of the carcinogen. Accordingly, they have promise as cancer prophylactics, including in combination with agents operating through other mechanisms. PMID- 12570343 TI - What is your diagnosis? Micro-angiopathic haemolytic anaemia secondary to splenic infarcts. PMID- 12570344 TI - Kinking of endotracheal tubes during maxima flexion of the atlanto-occipital joint in dogs. AB - Endotracheal tubes manufactured from red rubber and polyvinyl chloride (PVC) were compared, using radiographic techniques, in dog cadavers, under in vitro conditions and in anaesthetised dogs (in vivo study). All endotracheal tubes were radiographed in 'neutral' and flexed positions and the percentage reduction in the ventrodorsal radiographical diameter of the tube was calculated. The red rubber tubes kinked completely in 40 per cent of the cadavers, 75 per cent of the in vitro study and 40 per cent of anaesthetised dogs. The PVC tubes kinked completely in only one case (in vivo study). However, these tubes had a mean reduction in diameter of 15 per cent in the dog cadaver study, 19 per cent in the in vitro study and 26 per cent in the in vivo study. During procedures in which the trachea is intubated and the atlanto-occipital joint of the patient must be flexed, PVC endotracheal tubes may be less prone to kinking than red rubber tubes. PMID- 12570345 TI - Heritability and complex segregation analysis of hypoadrenocorticism in the standard poodle. AB - The heritability of hypoadrenocorticism (Addison's disease) was evaluated in 778 standard poodles with known Addisonian phenotypes. Addisonian status was confirmed clinically by adrenocorticotropic hormone (ACTH) challenge and 8.6 per cent of the poodles enrolled in the study were classified as being Addisonian. Hypoadrenocorticism affected both sexes with equal probability (P > 0.1). The most common coat colours had a negligible effect on the incidence of hypoadrenocorticism (P > 0.09), although red coat colour had a significant impact on the disease, probably due to the relatively small numbers of dogs with that coat colour. The heritability of hypoadrenocorticism in the standard poodle was estimated to be 0.75. Complex segregation analyses suggested that hypoadrenocorticism in the breed is influenced by an autosomal recessive locus. Clarification of both the heritability and mode of inheritance of hypoadrenocorticism in the standard poodle allows for better-informed breeding decisions. PMID- 12570346 TI - Renal abscess in a dog with transient diabetes mellitus. AB - A nine-year-old, intact female dalmatian with diabetes mellitus and a renal abscess is described. The renal abscess was treated surgically by nephrectomy, and the diabetes mellitus resolved with ovariohysterectomy. Abdominal ultrasound and ultrasound-guided aspiration of the abscess were helpful in establishing a diagnosis. To the authors' knowledge, this is the first report of a renal abscess in a dog with diabetes mellitus. PMID- 12570347 TI - Suprascleral removal of a foreign body from the retrobulbar muscle cone in two dogs. AB - The successful application of a suprascleral approach to remove a foreign body within the retrobulbar muscle cone in two dogs is described. The episcleral space was entered through a small opening in the paralimbal conjunctiva and bulbar vagina. Under ultrasound guidance, a curved haemostat with a fine tip was advanced along the scleral surface to the foreign body, which was then carefully grasped and removed using gentle traction. The conjunctival incision was left to heal secondarily. The suprascleral approach may allow the quick removal of non cutting and non-barbed foreign bodies from the episcleral space or the retrobulbar muscle cone, without the need for extensive tissue dissection. To reduce the risk of damaging extraocular tissues, the procedure must be performed cautiously and under ultrasound guidance. The globe should be continuously observed for any movement suggesting traction or pushing forces on bulbar or adjacent anatomical structures. PMID- 12570348 TI - Parotid salivary duct mucocoele and sialolithiasis following parotid duct transposition. AB - An 11-year-old, female neutered English springer spaniel presented with recurrent episodes of mucopurulent discharge from the right eye, 17 months after parotid duct transposition, culminating in the development of a swelling over the infraorbital area. A diagnosis of parotid duct obstruction with resulting rupture of the duct and sialocele was made. On exploratory surgery, three sialoliths were removed from various sites within the duct. Surgical repair of the duct restored normal saliva flow. PMID- 12570349 TI - Panniculitis associated with pancreatitis in a cocker spaniel. AB - A 13-year-old, male cocker spaniel presented with a history of inappetence, depression and reluctance to stand. The dog had multiple, ulcerated skin lesions which were diagnosed as panniculitis by histopathology. A diagnosis of pancreatitis was made on the basis of markedly elevated serum lipase concentrations, abdominal ultrasonography which showed an abnormal lobulated area of hypoechoic tissue in the body and right lobe of the pancreas, and a fine needle biopsy from this area which revealed large numbers of degenerate neutrophils. After treatment with antibiotics and prednisolone, the dog made a full clinical recovery and was free of clinical signs for four months. The dog was euthanased five months later and postmortem examination revealed chronic, active pancreatitis and a pancreatic adenoma. This is the first report of antemortem diagnosis of pancreatitis and panniculitis in a dog. PMID- 12570350 TI - Cardiology and imaging at Congress 2003. PMID- 12570351 TI - United by an enthusiasm for feline medicine. PMID- 12570352 TI - Current perspectives on the genetics of inherited diseases. PMID- 12570353 TI - Theoretically derived CVLT subtypes in HIV-1 infection: internal and external validation. AB - The present study sought to delineate empirically derived memory subtypes using the California Verbal Learning Test (CVLT; Delis et al., 1987) in a sample of adults with HIV-1 infection (N = 154). Confirmatory factor analysis was used to evaluate eight models of the CVLT structure suggested by Wiegner and Donders (1999). A four-factor model, consisting of Attention Span, Learning Efficiency, Delayed Recall, and Inaccurate Recall appeared to be the best fitting model. Variables with the highest factor loadings from the model were entered in a two stage cluster analysis. Four reliable CVLT clusters or subtypes were identified: Normal, Atypical, Subsyndromal, and Frontal-striatal. Internal and external validation of subtypes demonstrated that clusters were stable and clinically interpretable. Subtypes were meaningfully related to neuropsychological functioning, and to some extent, depressive symptomatology. Subtypes did not differ significantly with respect to subjective neurocognitive complaints and markers of HIV-1 disease. The present findings highlight the heterogeneity of memory profiles in HIV-1 infection and support a frontal-striatal conceptualization of verbal memory performance. The identification of robust HIV 1 memory subtypes may have important implications for the clinical management of adults infected with HIV-1 infection. PMID- 12570354 TI - Prodromal frontal/executive dysfunction predicts incident dementia in Parkinson's disease. AB - To identify the cognitive characteristics predictive of incident dementia in Parkinson's disease (PD), we examined the baseline neuropsychological profiles of 18 initially non-demented patients with PD who met diagnostic criteria for dementia (PDD) at one-year follow-up. PDD participants' baseline neuropsychological test scores were compared to the baseline performance of 18 patients with PD who did not meet criteria for dementia at one-year follow-up (PDND) and 18 normal controls (NC). The three groups were matched on baseline demographic and disease variables. Relative to the PDND group, the incident PDD participants demonstrated significantly poorer performance on digits backward (Wechsler Memory Scale-Revised), word list learning and recognition (California Verbal Learning Test), and perseverative errors on the Wisconsin Card Sorting Test. Each of these baseline neuropsychological variables exhibited adequate diagnostic classification accuracy in predicting PDD and PDND group membership at follow-up. These results suggest that subtle frontal/executive dysfunction is evident during the immediate PDD prodrome and may be of prognostic value in identifying PD patients at risk for dementia. Accordingly, neuropsychological evaluation may facilitate early identification of PDD and thereby inform appropriate dispositional planning. PMID- 12570355 TI - Dissociation of perceptual and motor inhibition processes through the use of novel computerized conflict tasks. AB - Efficient behavioral functioning requires early perceptual inhibition of irrelevant stimuli and later motor inhibition of inappropriate responses. The Perceptual and Motor Conflict Tasks were developed to differentially assess perceptual and motor inhibition, and to determine whether these processes utilize separate or shared cognitive resources. The computerized tasks include six subtests involving a box or an arrow appearing in various locations. Subjects respond by pressing a key on the left or right side of a keyboard. In different subtests, arrow direction or stimulus location determines correct responses. Perceptual inhibition assessment requires the subject to respond to a conflicting arrow direction while ignoring stimulus location. Motor inhibition assessment involves the subject responding in the direction opposite to that indicated by a centrally located arrow. In a neurologically normal sample (N = 44), reaction time analyses yielded significant Perceptual and Motor Conflict main effects, with slower performance under conflict conditions, but no significant Perceptual x Motor interaction. The lack of a significant Perceptual x Motor interaction, according to the additive factor model, indicates that these two processes utilize distinct cognitive resources. Nevertheless, performance on the two conflict tasks was significantly correlated with each other, and Perceptual Conflict performance was significantly correlated with Stroop interference. PMID- 12570356 TI - Ecological validity of a simplified version of the multiple errands shopping test. AB - Shallice and Burgess (1991) reported the utility of the Multiple Errands Test (MET) in discriminating executive deficits in three frontal lobe patients with preserved high IQ, who were otherwise unimpaired on tests of executive function. The aim of this study was to ascertain the value of a simplified version of the MET (MET-SV) for use with the range of people more routinely encountered in clinical practice. Main findings were as follows: 1) The test discriminated well between neurological patients and controls, and the group effects remained when the difference in current general cognitive functions (WAIS-R FSIQ) was taken into account. 2) The best predictors of performance in the healthy control group (n = 46) were age and the number of times participants asked for help (with more requests associated with poorer performance). 3) In the neurological group, two clear patterns of failure emerged, with performance either characterized by rule breaking or failure to achieve tasks. These two patterns were associated with different dysexecutive symptoms in everyday life. 4) The patients not only made more errors than controls, but also different ones. A scoring method that took this into account markedly increased test sensitivity. 5) Many patients passed traditional tests of executive frontal lobe function but still failed the MET-SV. This pattern was strongly associated with observed dysexecutive symptoms in everyday life. The results demonstrate the clinical utility of the test, and suggest that there are two common and independent sources of failure on multitasking tests in a general neurological population: memory dysfunction, and initiation problems. PMID- 12570357 TI - Clinical validity of the Chinese version of Mattis Dementia Rating Scale in differentiating dementia of Alzheimer's type in Hong Kong. AB - The present study aimed to examine the clinical validity and applicability of the Chinese version of Mattis Dementia Rating Scale (CDRS) for elderly Chinese individuals. The scale was found to have good reliability with internal consistency ranging from 0.7 to 0.9. Its significant correlation with the Chinese version of Mini-Mental State Examination (CMMSE) suggested satisfactory construct validity of the scale. The discriminant validity of the CDRS in differentiating Alzheimer's disease (AD) patients and Normal Control (NC) elderly was also supported by the Receiver Operating Characteristics (ROC) analysis. Several cut off points were presented for different clinical or research applications, and formulas to adjust for the age and educational level of the CDRS total score were provided. The Initiation/Perseveration and Memory subscales were suggested to be an abbreviated version of the CDRS for a quick screening. The present results also showed the association between the impairment of the CDRS performance and the progress of the AD. PMID- 12570358 TI - A consideration of neuropsychologically normal schizophrenia. AB - Neuropsychological deficits are considered by many to be core features of schizophrenia. However, about 20% of patients with schizophrenia appear to have normal neuropsychological function. This study investigates this subgroup by comparing a "neuropsychologically normal" schizophrenia group to a non schizophrenic, non-brain damaged patient comparison (PC) sample, and to patients with definitive brain damage who performed normally on neuropsychological testing. All patients completed the Halstead-Reitan Neuropsychological Test Battery and were classified as neuropsychologically normal or impaired using the Average Impairment Rating (AIR). In a sample of 113 patients with schizophrenia, 19.5% were classified as neuropsychologically normal. The brain damaged neuropsychologically normal group (BD-NN) consisted of 14.3% of 124 subjects. These groups were compared with a patient non-schizophrenic, non-brain damaged group who were selected on the basis of having an Average Impairment Rating in the neuropsychologically normal range. The neuropsychologically normal schizophrenic group performed less well than the non-brain damaged, non schizophrenic patient comparison group on a number of tests, indicating that patients in this group may not be completely neuropsychologically normal, and would be better characterized as "high-functioning" or near normal. The results are discussed in regard to possible neurobiological differences between neuropsychologically impaired and intact schizophrenic patients, and implications for course and outcome. PMID- 12570359 TI - Computerized reaction time battery versus a traditional neuropsychological battery: detecting HIV-related impairments. AB - In recent years, interest in the use of computerized neuropsychological (NP) assessment measures has increased. However, there are limited data regarding how performance on these measures relates to performance on more traditional, clinical instruments. In the present study, 82 HIV+ men, who were all believed on clinical grounds to have neurobehavioral impairment, completed a traditional NP battery (TNB) and the California Computerized Assessment Package (CalCAP, a collection of computerized reaction time tests). Summary scores based on a TNB, as well as those based on the CalCAP, demonstrated significant associations with both degree of immunosuppression (CD4 count) and detectable viral load in cerebrospinal fluid, but not with detectable viral load in plasma. Established norms on the TNB and CalCAP batteries resulted in classifying 57% and 49% of the HIV+ sample as impaired, respectively. When using the TNB as the "gold standard," impairment classifications based on CalCAP summary scores exhibited a sensitivity of 68% and a specificity of 77%. Overall agreement on impairment classifications between batteries was low (kappa = .44). Data from this study suggest that traditional NP batteries and computerized reaction time tests do not measure the same thing, and are not interchangeable in examining HIV-related NP impairments. PMID- 12570360 TI - Are psychophysical functions derived from line bisection reliable? AB - Psychophysical functions are used to characterize both normal perception and altered perception among patients with neglect, yet the reliability of these functions is rarely examined. The present study examined two-week, test-retest reliability for power functions derived from line bisection data among 58 normal, young and old, male and female subjects. Power function exponents and constants were, at best, moderately reliable over time. The size of the exponent tended to decrease at retesting. Reliability coefficients varied by age and gender; they were highly significant for young men, marginally significant for older men, and non-significant for women. Race influenced reliability as coefficients were significant for Caucasian subjects but not for African American subjects. Age and gender effects in this study parallel those in the literature on pseudoneglect, and they may reflect hemispheric differences in visuo-spatial processing, magnitude estimation, or both. PMID- 12570361 TI - Is there disproportionate impairment in semantic or phonemic fluency in schizophrenia? AB - Phonemic and semantic fluency involve the capacity to generate words beginning with particular letters or belonging to particular categories, respectively. The former has been associated with frontal lobe function and the latter with temporoparietal function, but neuroimaging studies indicate overlap of underlying neural networks. Schizophrenia patients may experience disproportionate semantic fluency impairment owing to abnormal semantic organization; however, executive dysfunction in schizophrenia suggests possible disproportionate phonemic fluency impairment. Moreover, little is known about the diagnostic specificity of either verbal fluency deficit to schizophrenia or their stability over time. We examined 83 schizophrenia patients, 15 bipolar disorder patients, and 83 normal controls. Both fluency types were impaired in schizophrenia patients. Schizophrenia patients as a whole manifested disproportionate semantic fluency impairment relative to bipolar disorder patients, but only a subset of schizophrenia patients manifested disproportionate semantic fluency impairment relative to controls. Few characteristics, except to some extent paranoid-nonparanoid subtype, meaningfully differentiated schizophrenia patients with and without this disproportionate impairment. Verbal fluency measures were moderately stable over a 4-year period in schizophrenia patients and controls (.48 < rs < .79). These results mirror a literature that overall suggests a small degree of disproportionate semantic fluency impairment in schizophrenia, but also some heterogeneity in fluency deficits. PMID- 12570362 TI - What does the WMS-III tell us about memory changes with normal aging? AB - The standardization sample from the WMS-III (N = 1250), which varied in age from 16 to 89, was used to determine whether encoding, retrieval, or storage of verbal and spatial information was most affected by normal aging. Immediate and delayed recall and recognition of Logical Memory and Visual Reproduction were examined. Immediate verbal and spatial recall significantly deteriorated with increasing age, and the age-associated deterioration in delayed recall and recognition was largely explained by poorer immediate memory. These findings, in concert with the smaller aging effects for percent retention after a delay, suggest that the aging effect is due to deterioration in encoding more than retrieval or storage of new information. While Visual Reproduction deteriorated more rapidly with age than Logical Memory, the pattern of performance decrements as a function of age were comparable across both tests. Decreases in performance were first seen in the fifth decade with gradual deterioration until the eighth decade when there was another precipitous drop. These results suggest that functions that are more dependent on the frontal lobes are more vulnerable to aging than those that are more dependent on the temporal lobes. PMID- 12570363 TI - Age and gender interactions on verbal memory performance. AB - Age and gender effects on verbal episodic memory are well established. However, the possibility of interactions between age and gender has been raised by studies linking estrogen and verbal memory performance, and by research suggesting gender differences in age-related cortical atrophy. We evaluated whether age by gender interactions in verbal memory were present. Subjects within three years of the median age of menopause were excluded from a large cohort of normal subjects, resulting in a younger sample (16-47 years) of 288 men and 285 women, and an older sample (55-89 years) of 201 men and 245 women. All subjects were administered the CVLT-2, a multiple-trial list-learning task. Verbal memory was negatively correlated with age for younger men, older men, and older women, but not for younger women. Multivariate analyses indicated age by gender interactions on memory for the younger group but not the older group. Results indicate that verbal memory declines with age for younger men but not younger women, whereas both older men and older women show age-related declines. These findings are consistent with hypotheses linking estrogen and verbal memory performance, and with imaging data suggesting that age-related hippocampal atrophy is found in younger men but not younger women. The role of estrogen on cognition in normal aging warrants further study. PMID- 12570364 TI - Frontal-striatal circuit functions: context, sequence, and consequence. AB - The exact role of the basal ganglia in both the motor and non-motor domains has proven elusive since it is virtually impossible to refer to its function in isolation of cortical, and especially frontal cortical circuits. The result is that we often speak of frontal-striatal circuits and functions but this still leaves us in the dark when trying to specify basal ganglia information processing. A critical review of the data from both basic science and clinical studies suggests that we should break down processing along a temporal continuum, including the domains of context, sequential information processing, and feedback or reinforcement (i.e., the consequences of action). This analysis would cut across other theoretical constructs, such as attention, central executive, memory, and learning functions, traditionally employed in the neuropsychological literature. Under specified behavioral constraint, the basal ganglia can then be seen to be involved in fundamental aspects of attentional control (often covert), in the guidance of the early stages of learning (especially reinforcement-based, but also encoding strategies in explicit paradigms), and in the associative binding of reward to cue salience and response sequences via dopaminergic mechanisms. Parkinson's disease is considered to offer only a limited view of basal ganglia function due to partial striatal depletion of dopamine and the potential involvement of other structures and transmitters in its pathology. It is hoped that the present formulation will suggest new heuristic research strategies for basal ganglia research, permitting a closer link to be established between neurophysiological, functional imaging and neuropsychological paradigms. PMID- 12570365 TI - Qualitative features of finger movement during the Halstead finger oscillation test following traumatic brain injury. AB - Qualitative and quantitative performance on the Halstead Finger Tapping test may help differentiate brain dysfunctional patients from normal controls. "Normal" and "abnormal" finger tapping patterns during this task have been characterized and illustrated pictorially. Data from 65 patients with traumatic brain injury (TBI) and 15 normal controls support the dual proposition that (1) abnormal finger tapping patterns are more commonly observed in TBI patients than in controls and (2) the frequency of abnormal finger movements may relate to the severity of TBI during the acute stages after trauma. Future prospective studies are needed to replicate these findings. PMID- 12570366 TI - Ernest Guenther award in chemistry of natural products. Amphibian skin: a remarkable source of biologically active arthropod alkaloids. PMID- 12570367 TI - Design and synthesis of 8-hydroxy-[1,6]naphthyridines as novel inhibitors of HIV 1 integrase in vitro and in infected cells. AB - Naphthyridine 7 inhibits the strand transfer of the integration process catalyzed by integrase with an IC50 of 10 nM and inhibits 95% of the spread of HIV-1 infection in cell culture at 0.39 microM. It does not exhibit cytotoxicity in cell culture at < or =12.5 microM and shows a good pharmacokinetic profile when dosed orally to rats. The antiviral activity of 7 and its effect on integration were confirmed using viruses with specific integrase mutations. PMID- 12570368 TI - Substituted pyrazolopyridopyridazines as orally bioavailable potent and selective PDE5 inhibitors: potential agents for treatment of erectile dysfunction. AB - Novel pyrazolopyridopyridazine derivatives have been prepared as potent and selective PDE5 inhibitors. Compound 6 has been identified as a more potent and selective PDE5 inhibitor than sildenafil (1). It is as efficacious as sildenafil in in vitro and in vivo PDE5 inhibition models, and it is orally bioavailable in rats and dogs. The superior isozyme selectivity of 6 is expected to exert less adverse effects in humans when used for erectile dysfunction treatment. PMID- 12570369 TI - Metabolism-directed optimization of 3-aminopyrazinone acetamide thrombin inhibitors. Development of an orally bioavailable series containing P1 and P3 pyridines. AB - Recent efforts in the field of thrombin inhibitor research have focused on the identification of compounds with good oral bioavailability and pharmacokinetics. In this manuscript we describe a metabolism-based approach to the optimization of the 3-(2-phenethylamino)-6-methylpyrazinone acetamide template (e.g., 1) which resulted in the modification of each of the three principal components (i.e., P1, P2, P3) comprising this series. As a result of these studies, several potent thrombin inhibitors (e.g., 20, 24, 25) were identified which exhibit high levels of oral bioavailability and long plasma half-lives. PMID- 12570370 TI - Structure-based de novo design, synthesis, and biological evaluation of non-azole inhibitors specific for lanosterol 14alpha-demethylase of fungi. AB - The active site of lanosterol 14alpha-demethylase (CYP51) was investigated via MCSS functional group mapping and LUDI calculations. Several non-azole lead molecules were obtained by coupling structure-based de novo design with chemical synthesis and biological evaluation. All of the lead molecules exhibited a strong inhibitory effect on CYP51 of Candida albicans. They occupy the substrate-binding site and interfere with the binding of azole antifungal agents in a competitive manner. The mode of action of the lead molecules was validated by spectrophotomeric analysis and SAR studies. This is the first successful example reported for the inhibitor design of the cytochrome P450 superfamily using the de novo design strategy. Because the affinity of the lead molecules for CYP51 was mainly attributed to their nonbonding interaction with the apoprotein, the studies presented here afford the opportunity to develop novel antifungal agents that specifically interact with the residues in the active site and avoid the serious toxicity arising from coordination binding with the heme of mammalian P450s. PMID- 12570371 TI - Identification, synthesis, and activity of novel blockers of the voltage-gated potassium channel Kv1.5. AB - The voltage-gated potassium channel Kv1.5 is regarded as a promising target for the development of new atrial selective drugs with fewer side effects. In the present study the discovery of ortho,ortho-disubstituted bisaryl compounds as blockers of the Kv1.5 channel is presented. Several compounds of this new class were synthesized and screened for their ability to block Kv1.5 channels expressed in Xenopus oocytes. The observed structure-activity relationship (SAR) is described by a pharmacophore model that consists of three hydrophobic centers in a triangular arrangement. The hydrophobic centers are matched by a phenyl or pyridyl ring of the bisaryl core and both ends of the side chains. The most potent compounds (e.g., 17c and 17o) inhibited the Kv1.5 channel with sub micromolar half-blocking concentrations and displayed 3-fold selectivity over Kv1.3 and no significant effect on the HERG channel and sodium currents. In addition, compounds 17c and 17m have already shown antiarrhythmic effects in a pig model. PMID- 12570372 TI - Surflex: fully automatic flexible molecular docking using a molecular similarity based search engine. AB - Surflex is a fully automatic flexible molecular docking algorithm that combines the scoring function from the Hammerhead docking system with a search engine that relies on a surface-based molecular similarity method as a means to rapidly generate suitable putative poses for molecular fragments. Results are presented evaluating reliability and accuracy of dockings compared with crystallographic experimental results on 81 protein/ligand pairs of substantial structural diversity. In over 80% of the complexes, Surflex's highest scoring docked pose was within 2.5 A root-mean-square deviation (rmsd), with over 90% of the complexes having one of the top ranked poses within 2.5 A rmsd. Results are also presented assessing Surflex's utility as a screening tool on two protein targets (thymidine kinase and estrogen receptor) using data sets on which competing methods were run. Performance of Surflex was significantly better, with true positive rates of greater than 80% at false positive rates of less than 1%. Docking time was roughly linear in number of rotatable bonds, beginning with a few seconds for rigid molecules and adding approximately 10 s per rotatable bond. PMID- 12570373 TI - 3-(4-Aroyl-1-methyl-1H-2-pyrrolyl)-N-hydroxy-2-alkylamides as a new class of synthetic histone deacetylase inhibitors. 1. Design, synthesis, biological evaluation, and binding mode studies performed through three different docking procedures. AB - Recently we reported a novel series of hydroxamates, called 3-(4-aroyl-1H-2 pyrrolyl)-N-hydroxy-2-propenamides (APHAs), acting as HDAC inhibitors (Massa, S.; et al. J. Med. Chem. 2001, 44, 2069-2072). Among them, 3-(4-benzoyl-1-methyl-1H-2 pyrrolyl)-N-hydroxy-2-propenamide 1 was chosen as lead compound, and its binding mode into the modeled HDAC1 catalytic core together with its histone hyperacetylation, antiproliferative, and cytodifferentiating properties in cell based assays were investigated (Mai, A.; et al. J. Med. Chem. 2002, 45, 1778 1784). Here we report the results of some chemical manipulations performed on (i) the aroyl portion at the C4-pyrrole position, (ii) the N(1)-pyrrole substituent, and (iii) the hydroxamate moiety of 1 to determine structure-activity relationships and to improve enzyme inhibitory activity of APHAs. In the 1 structure, pyrrole N(1)-substitution with groups larger than methyl gave a reduction in HDAC inhibiting activity, and replacement of hydroxamate function with various non-hydroxamate, metal ion-complexing groups yielded poorly active or totally inactive compounds. On the contrary, proper substitution at the C4 position favorably affected enzyme inhibiting potency, leading to 8 (IC50 = 0.1 micro M) and 9 (IC50 = 1.0 micro M) which were 38- and 3.8-fold more potent than 1 in in vitro anti-HD2 assay. Against mouse HDAC1, 8 showed an IC50 = 0.5 micro M (IC50 of 1 = 4.9 micro M), and also in cell-based assay, 8 was endowed with higher histone hyperacetylating activity than 1, although it was less potent than TSA and SAHA. Such enhancement of inhibitory activity can be explained by the higher flexibility of the pyrrole C4-substituent of 8 which accounts for a considerably better fitting into the HDAC1 pocket and a more favorable enthalpy ligand receptor energy compared to 1. The enhanced fit allows a closer positioning of 8 hydroxamate moiety to the zinc ion. These findings were supported by extensive docking studies (SAD, DOCK, and Autodock) performed on both APHAs and reference drugs (TSA and SAHA). PMID- 12570374 TI - Antineoplastic agents. 487. Synthesis and biological evaluation of the antineoplastic agent 3,4-methylenedioxy-5,4'-dimethoxy-3'-amino-Z-stilbene and derived amino acid amides. AB - An efficient synthesis of 3,4-methylenedioxy-5,4'-dimethoxy-3'-amino-Z-stilbene (1c) and hydrochloride (1d) is reported. The nitrostilbene intermediate 6a was obtained via a Wittig reaction using phosphonium salt 4 and 3-nitro-4 methoxybenzaldehyde 5. A one-step reduction using zinc in acetic acid produced the synthetic objective amine 1c. The coupling of this amine with various Fmoc amino acids, followed by cleavage of the alpha-amine protecting group, resulted in a series of new cancer cell growth inhibitory amides. Amine 1c, hydrochloride 1d, glycine amide 3b, and tyrosine amide 3f had the highest level (GI50 = 10(-2) 10(-3) micro g/mL) of activity against a panel of six human and one animal (P388) cancer cell lines. Amine 1c and its hydrochloride 1d potently inhibited tubulin polymerization by binding at the colchicine site, while the amides had little activity against purified tubulin. Nevertheless, most of the amides caused a marked increase in the mitotic index of treated cells, indicating that tubulin was their intracellular target. PMID- 12570375 TI - 4-Substituted 4-hydroxycyclohexa-2,5-dien-1-ones with selective activities against colon and renal cancer cell lines. AB - The synthesis and antitumor evaluation of a series of new heteroaromatic- and aromatic-substituted hydroxycyclohexadienones ("quinols"), and their imine counterparts, are described. The quinols were synthesized via the addition of a lithiated aromatic moiety to a quinone ketal followed by deprotection. When the aromatic portion of the molecule is a fused heterobicyclic structure (e.g., benzothiazole derivative 7a), potent in vitro antitumor activity was observed in HCT 116 (GI50 = 40 nM) and HT 29 (GI50 = 380 nM) human colon as well in as MCF-7 and MDA 468 human breast cancer cell lines. When examined on the NCI Developmental Therapeutics Screening Program in vitro screen (60 human cancer cell lines), active compounds in this series consistently displayed a highly unusual pattern of selectivity; cytotoxicity (LC50) was concentrated in certain colon and renal cell lines only. Analogue 7a also showed in vivo antitumor activity against human RXF 944XL renal xenografts in nude NMRI mice and is the focus of further study. PMID- 12570376 TI - Synthesis and in vitro and in vivo antimalarial activity of N1-(7-chloro-4 quinolyl)-1,4-bis(3-aminopropyl)piperazine derivatives. AB - Three series of monoquinolines consisting of a 1,4-bis(3-aminopropyl)piperazine linker and a large variety of terminal groups were synthesized. Our aim was to prove that in related bisquinoline, it is the second quinoline moiety that is responsible for cytotoxicity and that it is not an absolute requirement for overcoming resistance to chloroquine (CQ). Eleven compounds displayed a higher selectivity index (ratio CC50/IC50 activity) than CQ, and one of them cured mice infected by Plasmodium berghei. PMID- 12570377 TI - Absorption classification of oral drugs based on molecular surface properties. AB - The aim of this study was to investigate whether easily calculated and comprehended molecular surface properties can predict drug solubility and permeability with sufficient accuracy to allow theoretical absorption classification of drug molecules. For this purpose, structurally diverse, orally administered model drugs were selected from the World Health Organization (WHO)'s list of essential drugs. The solubility and permeability of the drugs were determined using well-established in vitro methods in highly accurate experimental settings. Descriptors for molecular surface area were generated from low-energy conformations obtained by conformational analysis using molecular mechanics calculations. Correlations between the calculated molecular surface area descriptors, on one hand, and solubility and permeability, on the other, were established with multivariate data analysis (partial least squares projection to latent structures (PLS)) using training and test sets. The obtained models were challenged with external test sets. Both solubility and permeability of the druglike molecules could be predicted with high accuracy from the calculated molecular surface properties alone. The established correlations were used to perform a theoretical biopharmaceutical classification of the WHO-listed drugs into six classes, resulting in a correct prediction for 87% of the essential drugs. An external test set consisting of Food and Drug Administration (FDA) standard compounds for biopharmaceutical classification was predicted with 77% accuracy. We conclude that PLS models of easily comprehended molecular surface properties can be used to rapidly provide absorption profiles of druglike molecules early on in drug discovery. PMID- 12570378 TI - Design, synthesis, and biological evaluation of a series of fluoroquinoanthroxazines with contrasting dual mechanisms of action against topoisomerase II and G-quadruplexes. AB - Topoisomerase inhibitors are important and clinically effective drugs, while G quadruplex-interactive compounds that disrupt telomere maintenance mechanisms have yet to be proven useful in the clinic. If G-quadruplex-interactive compounds are to be clinically useful, it will most likely be in combination with more established cytotoxic agents. We have previously reported on a family of topoisomerase II inhibitors that also interact with G-quadruplexes. On the basis of previously established structure-activity relationships (SARs) for compounds that are able to inhibit topoisomerase II or interact with G-quadruplex to varying degrees, we have now designed and synthesized four new fluoroquinoanthroxazines (FQAs) that have different profiles of mixed topoisomerase II poisoning effects and G-quadruplex interactions. The biological profiles of the four new compounds were determined with respect to G-quadruplex interaction (polymerase stop and photocleavage assays) and topoisomerase II interaction (DNA cleavage and kDNA decatenation assays), alongside cytotoxicity tests with matched pairs of topoisomerase II-resistant and topoisomerase II sensitive cells and with telomerase (+) and ALT (+) cell lines (ALT = alternative lengthening of telomeres). From this study, we have identified two FQAs with sharply contrasting profiles of potent G-quadruplex interaction with a weak topoisomerase II poisoning effect, and vice versa, for further evaluation to determine the optimum combination of these activities in subsequent in vivo studies. PMID- 12570379 TI - Orally active analogues of the dopaminergic prodrug 6-(N,N-di-n-propylamino) 3,4,5,6,7,8-hexahydro-2H-naphthalen-1-one: synthesis and pharmacological activity. AB - A series of analogues of 6-(N,N-di-n-propylamino)-3,4,5,6,7,8-hexahydro-2H naphthalen-1-one (6), an enone prodrug of the mixed DA D(1)/D(2) agonist 5,6-diOH DPAT (2), was synthesized. The pharmacological profiles of these new enones and their in vivo pharmacological activities were investigated in the Ungerstedt rat rotation model for Parkinson's disease. At 0.1 mg kg(-1) po, the N-methyl-N-n propyl (12) and the N-ethyl-N-propyl (13) analogues induced pronounced and long lasting pharmacological effects. The pharmacological profile of enone 12 was found to be similar to that of 6, while enone 13 was significantly more potent than 6 (p < 0.01). Analyses of rat brains after the administration of (-)-6 and 13 indicated the presence of hydroxylated metabolites of the parent enones. It is speculated that such metabolites are alpha'-hydroxylated enones that may constitute the first step in the formation of the corresponding catechols. PMID- 12570380 TI - Design, synthesis, and biological activities of classical N-[4-[2-(2-amino-4 ethylpyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-l-glutamic acid and its 6-methyl derivative as potential dual inhibitors of thymidylate synthase and dihydrofolate reductase and as potential antitumor agents. AB - Two novel analogues, N-[2-amino-4-ethyl[(pyrrolo[2,3-d]pyrimidin-5 yl)ethyl]benzoyl]-l-glutamic acid (2) and N-[2-amino-4-ethyl-6 methyl[(pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-l-glutamic acid (4), were designed and synthesized as potent dual inhibitors of thymidylate synthase (TS) and dihydrofolate reductase (DHFR) and as antitumor agents. Compound 2 had inhibitory potency against human DHFR similar to N-[4-[2-(amino-3,4-dihydro-4-oxo 7H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-L-glutamic acid (LY231514) and 1, whereas 4 was inactive against human DHFR. Both 2 and 4 were more potent than LY231514 against E. coliTS. Against human TS, 2 was 7-fold less potent than LY231514 and 4 showed similar inhibitory activity as LY231514. In contrast to 2, which was an efficient substrate of human folypolyglutamate synthetase (FPGS), 4 was a poor substrate of FPGS. Compound 2 showed GI50 values in the nanomolar range against more than 18 human tumor cell lines in the standard NCI preclinical in vitro screen. PMID- 12570381 TI - Preparation of 7-substituted ginkgolide derivatives: potent platelet activating factor (PAF) receptor antagonists. AB - Ginkgolides are structurally unique constituents of Ginkgo biloba extracts and are known antagonists of the platelet-activating factor (PAF) receptor. Ginkgolide C is 25-fold less potent than ginkgolide B as a PAF receptor antagonist, due to the presence of the 7beta-OH. Recently, we found that 7alpha fluoro ginkgolide B was equipotent to ginkgolide B underlining the critical importance of the 7-position of ginkgolides for PAF receptor activity. Herein we describe the synthesis of a series of ginkgolide B derivatives with modifications at the 7-position and the pharmacological evaluation of these derivatives as assayed by cloned PAF receptors. In two cases nucleophilic attack on a 7beta-O triflate ginkgolide B did not lead to the expected products, but gave rise to two unprecedented ginkgolide derivatives, one with a novel rearranged skeleton. Furthermore, standard reduction of 7alpha-azido ginkgolide B did not give the expected primary amine, but instead yielded alkylated amines depending on the solvent employed. Pharmacological testing with cloned PAF receptors showed that ginkgolides with 7alpha-substitutents had increased affinity compared to 7beta substituents, in particular 7alpha-chloro ginkgolide B, the most potent nonaromatic ginkgolide derivative described to date with a K(i) value of 110 nM. PMID- 12570382 TI - Syntheses and antiproliferative activities of 7-azarebeccamycin analogues bearing one 7-azaindole moiety. AB - Rebeccamycin analogues containing one azaindole unit, with and without a methyl group on the imide nitrogen and with the sugar moiety coupled either to the indole nitrogen or to the azaindole nitrogen were synthesized. To increase the solubility and induce stronger interactions with the target macromolecules, a bromo or nitro substitutent was introduced on the indole unit. The DNA binding and topoisomerase I inhibition properties were investigated together with the antiproliferative activities toward nine tumor cell lines. In addition, the effect of the compounds on the cell cycle of L1210 leukemia cells was examined. The nonaza analogues were found to be cytotoxic against all cell lines of the panel whereas the aza-analogues showed a selective action toward certain cell lines. They strongly inhibited the proliferation of SK-N-MC neuroblastoma, A431 epidermoid carcinoma and NCI-H69 small cell lung carcinoma cells, but showed little or no cytotoxic effect against IGROV ovary carcinoma, HT29 colon carcinoma, and A549 non small cell lung carcinoma cells. Whatever their cytotoxicity profile, all compounds induce similar cell cycle effects, with a marked G2+M block observed with L1210 leukemia cells. The data suggest that the molecular mechanism of action of the aza-analogue derivatives is different from that of rebeccamycin. PMID- 12570383 TI - 3-(alphaR)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N alkyl-N-arylbenzamides: potent, non-peptidic agonists of both the micro and delta opioid receptors. AB - Opioid analgesics with both micro and delta opioid receptor activation represent a new approach to the treatment of severe pain with an improved safety profile. Compounds with this profile may exhibit strong analgesic properties due to micro agonism, with a reduced side effect profile resulting from delta agonism. Replacing the p-diethylamide of the known potent delta opioid receptor selective agonist BW373U86 with a m-diethylamide resulted in a compound with agonist activity at both the micro and delta opioid receptors. Modifying the amide to an N-methyl-N-phenylamide increased agonist potency at both receptors. A series of 3 (alphaR)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N alkyl-N-arylbenzamides have been made to explore the structure-activity relationship (SAR) around the N-methyl-N-phenylamide. Several potent agonists of both the micro and delta opioid receptors have been identified, including (+)-3 ((alphaR)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N (4-fluorophenyl)-N-methylbenzamide (23), which has EC50 values of 0.67 and 1.1 nM at the micro (guinea pig ileum assay) and delta (mouse vas deferens assay) opioid receptors, respectively. PMID- 12570384 TI - CC-1065 analogues bearing different DNA-binding subunits: synthesis, antitumor activity, and preliminary toxicity study. AB - CC-1065 analogues bearing different DNA-binding subunits were synthesized. A terminal C5-NO2 and -F moiety at the DNA-binding subunit increased the drug's potency and antitumor efficacy. A C5-OCH3 reduced the potency and antitumor efficacy. Compound (+/-)-7, bearing a trans double bond, had increased antitumor efficacy. A preliminary toxicity study indicated that terminal C5-OCH3 and acetamido moieties at the DNA-binding subunit caused delayed death in mice. PMID- 12570385 TI - New semisynthetic quassinoids with in vivo antimalarial activity. AB - On the basis of a comparative analysis for stability in mouse serum between 15-O acetylbruceolide and bruceolide 15-methyl carbonate, several 3,15-dialkyl carbonates of bruceolide were synthesized and their in vitro antimalarial activity was assessed. Methyl, ethyl, and isopropyl carbonates with pronounced in vitro activity were further evaluated for in vivo antimalarial potency. Both the methyl and ethyl carbonates significantly increased the life span of mice as compared with 3,15-di-O-accetylbruceolide and chloroquine. PMID- 12570386 TI - Synthesis, structure-activity relationship, and evaluation of SR141716 analogues: development of central cannabinoid receptor ligands with lower lipophilicity. AB - Exploration of the central CB1 cannabinoid receptors using positron emission tomography (PET) will allow for an understanding of the pharmacological and physiological role played by these receptors in the CNS. Current tracers are highly lipophilic compounds that exhibit very high nonspecific to specific binding ratios and as a result are inapt for use in humans. We have synthesized a series of less lipophilic analogues of SR141716 to serve as potential radioligands. Binding affinities of the series and a functional electrophysiological assay of three of our compounds have been presented. PMID- 12570387 TI - Synthesis and structure-affinity relationships of 1-[omega-(4-aryl-1 piperazinyl)alkyl]-1-aryl ketones as 5-HT(7) receptor ligands. AB - Structural requirements for 5-HT(7) receptor affinity and selectivity over that for the 5-HT(1A) receptor were studied on a series of 1-[omega-(4-aryl-1 piperazinyl)alkyl]-1-aryl ketones. The presence of a hydroxy or methoxy substituent on aryl ketone moiety, alkyl chain length, and the nature of N-1 piperazine substituent were explored. 6-[4-(3-Benzisoxazolyl)-1-piperazinyl]-1-(2 hydroxyphenyl)-1-hexanone (40) and its methoxy analogue 43 exhibited high 5-HT(7) receptor affinities (Ki = 2.93 nM and 0.90 nM, respectively) and agonist properties when tested for 5-HT(7) receptor-mediated relaxation of substance P induced guinea-pig ileum contraction. PMID- 12570390 TI - Monitoring of acid-base status of workers at a methyl methacrylate and polymethyl methacrylate production plant in Bulgaria. AB - This study was carried out on 104 workers at three work operations and a control (nonproduction) area, within a methyl methacrylate (MMA)/polymethyl methacrylate (PMMA) production facility in Bulgaria. Airborne monitoring was conducted over a 10-year period for MMA and the reactant chemicals methanol and acetone cyanhydrine at the MMA operation, and MMA was monitored at the PMMA operation. Acid-base status of the workers was evaluated using traditional criteria (pH, pCO(2), pO(2), and HCO(3) in plasma). Data from retrospective monitoring of air levels of the chemicals were compared with the acid-base status of workers at the plant. In some cases air concentrations exceeded the threshold limit value, with the highest percentage of overexposure occurring with airborne MMA in the PMMA production operation. Acid-base disruption indicated by reductions in plasma pH and HCO(3) was found for all groups except the control population. The highest percentage reduction was associated with PMMA production workers. Additionally, respiratory acidosis, indicated by increased pCO(2), was noted in the MMA production and maintenance groups, implying that the response to MMA exposure may involve both the metabolic and respiratory acidosis component. This study was unique in that the combined exposure to MMA and the precursor chemical (methanol) were shown to produce the same effects in workers. It is suggested that when combined exposure occurs, disruption of acid-base status may occur. Enforcement of PPM requirements for coveralls and gloves should prevent skin contamination. Additionally, improvement of equipment in MMA and PMMA production areas is recommended: (1) automation of some manual operations; (2) use of respiratory protection during equipment cleaning; and (3) installation of local ventilation when applicable. PMID- 12570391 TI - Contribution of toxicokinetic modeling to the adjustment of exposure limits to unusual work schedules. AB - This study compared two toxicokinetic approaches for determining correction factors to be applied to occupational exposure limits (ELs) for unusual exposure scenarios: a classic one-compartment toxicokinetic approach and the physiologically based toxicokinetic (PBTK) approach. The approaches were applied to three typical unusual exposure scenarios: four consecutive 10-hour workdays followed by 3 days of recovery; three consecutive 12-hour workdays followed by 4 days of recovery; and a 4/3 work schedule. Results indicate that use of an adjustment method for ELs based on contaminant toxicokinetics generates less protective correction factors (i.e., a smaller adjustment) than those obtained using the U.S. Occupational Safety and Health Administration approach, which is based on Haber's law. Among all scenarios tested, the highest adjustment required, resulting from the use of a toxicokinetic approach (PBTK or one compartment), was for the 4/3 work schedule and for a contaminant with a half life equal to 18 hours. In that case the ELs would need to be reduced by 26%. Based on previous work, the authors believe an adjustment based on a toxicokinetic approach is more realistic from a toxicological standpoint. Given the value of a substance's half-life, the use of the graphs of Hickey and Reist (developed from a one-compartment toxicokinetic model) is a rapid and reliable means of establishing the correction factor. However, this approach is limited to simple and repetitive scenarios. For more complex exposure scenarios, such as that corresponding to a 4/3 work schedule, a one-compartment model also can be developed for each of the needs. Finally, the use of PBTK models allows greater flexibility for adjusting ELs for novel work schedules. PMID- 12570392 TI - Control of wake-induced exposure using an interrupted oscillating jet. AB - A problem may arise in ventilation design when the contaminant source is located in the worker's wake, where turbulence and vortex formation can carry the contaminant into the breathing zone even though the source is downwind. It was found previously that forced directional variations in the flow can reduce or eliminate the vortex formation that causes these local reversals. Reported here is a simple realization of this concept, in which an oscillating jet of air was directed at a mannequin in an otherwise steady flow of air. A 50th percentile male mannequin was placed in a nearly uniform flow of approximately 0.18 m/sec (36 ft/min). A low-velocity tracer gas source (isobutylene) was held in the standing mannequin's hands with the upper arms vertical and the elbows at 90 degrees. Four ventilation scenarios were compared by concentration measurements in the breathing zone, using photoionization detectors: (A) uniform flow; (B) addition of a steady jet with initial velocity 5.1 m/sec (1.0 x 10(3) ft/min) directed at the mannequin's back, parallel to the main flow; (C) making the jet oscillate to 45 degrees on either side of the centerline with a period of 13 sec; and (D) introducing a blockage at the centerline so the oscillating jet never blew directly at the worker. At the 97.5% confidence level the interrupted oscillating jet (case D) achieved at least 99% exposure reduction compared with the uniform flow by itself (case A), at least 93% compared with the steady jet (case B), and at least 45% exposure reduction compared with the unblocked oscillating jet (case C). PMID- 12570393 TI - Comparison of coal mine dust size distributions and calibration standards for crystalline silica analysis. AB - Since 1982 standard calibration materials recommended for respirable crystalline silica analysis by the Mine Safety and Health Administration (MSHA) P7 Infrared Method and the National Institute for Occupational Safety and Health (NIOSH) X ray Diffraction (XRD) Analytical Method 7500 have undergone minor changes in size distribution. However, a critical assumption has been made that the crystalline silica in ambient mine atmosphere respirable dust samples has also remained essentially unchanged in particle size distribution. Therefore, this work compared recent particle size distributions of underground coal mine dust and the silica component of these dusts with estimated aerodynamic particle size distributions of calibration standard materials MIN-U-SIL 5, Berkeley 5, and SRM 1878 used by two crystalline silica analysis techniques. Dust impactor sampling data for various locations in 13 underground coal mines were analyzed for the respirable mass median aerodynamic diameters. The data suggest that the MSHA P7 method will underestimate the silica content of the sample by at most 7.4% in the median size range 0.9 to 3.6 microm, and that it is unlikely one would obtain any significant error in the MSHA P7 method analysis when the method uses Berkeley 5, MIN-U-SIL 5, or SRM 1878 as a calibration standard material. The results suggest that the NIOSH Analytical Method 7500 would be more appropriate for a dust sample that is representative of the total (no cyclone classifier) rather than the respirable airborne dust, particularly because the mass fraction in the size range below 4 microm is usually a small percentage of the total airborne dust mass. However, NIOSH Analytical Method 7500 is likely to underestimate the silica content of an airborne respirable dust sample by only 5 to 10%. The results of this study also suggest that any changes that may have occurred in the median respirable size of airborne coal mine dust are not significant enough to cause any appreciable error in the current methods used for respirable crystalline silica analysis. PMID- 12570394 TI - A method for detecting fungal contaminants in wall cavities. AB - This article describes a practical method for detecting the presence of both fungal spores and culturable fungi in wall cavities. Culturable fungi were collected in 25 mm cassettes containing 0.8 microm mixed cellulose ester filters using aggressive sampling conditions. Both culturable fungi and fungal spores were collected in modified slotted-disk cassettes. The sample volume was 4 L. The filters were examined microscopically and dilution plated onto multiple culture media. Collecting airborne samples in filter cassettes was an effective method for assessing wall cavities for fungal contaminants, especially because this method allowed the sample to be analyzed by both microscopy and culture media. Assessment criteria were developed that allowed the sample results to be used to classify wall cavities as either uncontaminated or contaminated. As a criterion, wall cavities with concentrations of culturable fungi below the limit of detection (LOD) were classified as uncontaminated, whereas those cavities with detectable concentrations of culturable fungi were classified as contaminated. A total of 150 wall cavities was sampled as part of a field project. The concentrations of culturable fungi were below the LOD in 34% of the samples, whereas Aspergillus and/or Penicillium were the only fungal genera detected in 69% of the samples in which culturable fungi were detected. Spore counting resulted in the detection of Stachybotrys-like spores in 25% of the samples that were analyzed, whereas Stachybotrys chartarum colonies were only detected on 2% of malt extract agar plates and on 6% of corn meal agar plates. PMID- 12570395 TI - Evaluation of three methodologies for assessing work activity during computer use. AB - The overall goal of this study was to evaluate three separate methodologies for gathering work activity information among computer users. These methodologies included worker self-report, work sampling, and activity monitoring. A repeated measures design was employed whereby data were collected simultaneously on each subject (n = 51) across three consecutive workdays. Exposure information gathered included keying time, mouse usage, and time spent performing various work tasks (i.e., writing, proofreading, handling documents). Subjects were recruited to represent a wide range of keyboard activity and mouse usage. The study found that worker self-reports overestimated actual keyboard usage by a factor of approximately 1.5 for workers using the keyboard an average of 4 hours per day to a factor of 4 for workers using the keyboard an average of 30 min per day. On average, there was an approximate twofold difference between worker self-reported keying time and that obtained via activity monitoring and work sampling. This trend was similar with regard to time spent using the computer mouse. Worker self reported mouse usage was approximately twofold higher than that obtained via activity monitoring or work sampling. Self-reported exposure information not only resulted in different estimates, but showed greater variance compared with the other methodologies. The results of this study suggest that the use of worker self-reported exposure information on keying time and mouse usage may not represent an accurate account of time spent performing these tasks. In the context of epidemiological studies work sampling and/or activity monitoring would be more suitable methodologies for obtaining such information. PMID- 12570396 TI - Postural stability is compromised by fatiguing overhead work. AB - In a laboratory setting, 16 participants performed a repetitive overhead tapping task for 3 hours or until self-terminated due to substantial shoulder discomfort. Several measures of postural sway and stability were obtained using a force plate, both during quiet standing and during performance of the tapping task. Sway area and peak sway velocity showed consistent increases with time, whereas changes in average velocity and peak whole-body center-of-mass acceleration were either small or nonsignificant. Although relatively insensitive to several task variables, changes in sway areas and peak velocities were substantially larger in trials terminated by the participants. It is argued that fatigue plays a more important role than simple task duration in causing the observed increases in sway, and hence decreases in postural stability. Potential whole-body consequences of localized musculoskeletal stresses appear supported by the results, and implications for safety, risks of falls, and work scheduling are discussed. PMID- 12570397 TI - Prevention strategies in industrial hygiene: a critical literature review. AB - Little is known of the extent of use of industrial hygiene prevention and control strategies in actual workplaces. The recent occupational safety and health literature was identified as a potential source for identifying which strategies are being utilized and evaluated as a means of controlling or preventing chemical hazards. Using preestablished selection criteria, the peer-reviewed industrial hygiene literature 1994-1999 was searched for articles describing prevention and control strategies for chemical hazards in actual workplaces. Ninety-two articles were found and categorized by the type of strategy discussed, whether strategies were implemented and evaluated, and by several other categories. Almost three quarters discussed engineering control strategies, mostly local exhaust ventilation. Administrative strategies, including housekeeping, personal hygiene, and medical surveillance, appeared in about half the articles. Personal protective equipment was considered in one-third of the articles, and primary prevention strategies, such as material substitution, were considered in one quarter. Intervention effectiveness was not consistently evaluated in these articles. In response to these findings, recommendations are made to improve the evaluation and promotion of effective prevention and control strategies. PMID- 12570399 TI - Quantifying synthetic vitreous fiber surface contamination in office buildings. AB - A reliable method for determining the distribution of synthetic vitreous fiber (SVF) densities found on building surfaces is presented with data collected from rooms in 40 office buildings in the United States. The initial survey of 20 buildings-complete with collection, counting, and analysis of SVF loading-found that nearly 60% of the samples had zero or one fiber counted. To resolve the nondetects, especially for lighter dust loading, the counting technique was modified. The detection limit improved as the zero fiber count decreased from about 43% in the initial survey to about 14% of the samples taken from 20 additional buildings. Analysis of the results indicated that the variability within rooms was much larger than the variability between rooms or between buildings. This variability is attributable to differences among the contact surfaces. Surfaces seldom contacted had greater SVF loading than those in frequent or constant contact. The technique developed will standardize evaluation of SVFs in building dust and can be used to assess potential causes of cutaneous and mucous membrane irritation symptoms in future studies. PMID- 12570398 TI - Hepatotoxicity associated with overexposure to 1,1-dichloro-2,2,2-trifluoroethane (HCFC-123). AB - 1,1-Dichloro-2,2,2-trifluoroethane (HCFC-123) was evaluated as a substitute for trichlorofluoromethane (CFC-11), and it appeared that a permissible exposure limit of 50 ppm was justified. When HCFC-123 was introduced as a precision cleaning agent in a controlled operation, marked elevations in serum alanine transaminase and serum aspartase transaminase were noted in exposed workers. Sampling taken during start-up documented personal samples from 24-480 ppm (375 and 21 min, respectively) and area samples of 18-180 ppm (375 and 21 min, respectively). Personal and area samples collected after the liver abnormalities were identified ranged from 5-12 ppm. Exposure data were not available for the period when the abnormalities are suspected to have developed. Two models were developed to estimate exposure during the unmonitored period: (1) the entire plant as a homogenous box and (2) evaporation into smaller work zones. Modeling using the entire building estimated 8-hour time-weighted average (TWA) exposures of 10-35 ppm. Modeled estimates of work area and air exchange rates indicated that degreaser exposed workers could have experienced peak levels of 280-2,100 ppm (8-hour TWAs 252-1,630 ppm). Modeling of the work environment, estimated to be one-third of the volume of the entire open building, indicated peak exposures of 28-210 ppm (8-hour TWAs 25-163 ppm). These ranges estimate the minimum and maximum exposure levels. The best estimates, using 12 air changes per day, suggest peak levels around the degreaser of 635-2,100 ppm (8-hour TWA 499-1,630 ppm) and 63-207 ppm (8-hour TWAs 50-163 ppm) in the work area. These are the first estimates of exposure level associated with these hepatotoxic effects; all are significantly higher than personal and area samples collected for HCFC-123 after the liver abnormalities were identified. PMID- 12570400 TI - Bactericidal treatment of raw cotton as the method of byssinosis prevention. AB - In early studies, research to control byssinosis focused on methods to reduce the trash in the textile mill environment. Dust control has been effective in reducing the prevalence of byssinosis, but simple reduction in dust levels does not always assure its prevention. Also, bacteria and fungi present in cotton do not in themselves cause byssinosis, but the endotoxins-heat-stable lipopolysaccharide-protein complexes contained in the cell wall of Gram-negative bacteria-are responsible for the development of this respiratory disease of workers on cotton, flax, and some other fibers. Experimental work was carried out in cotton fields in different cotton growing countries. Opened cotton capsules were treated by spraying them with bactericidal water solutions of benzododecinium bromide to avoid the growth of bacteria by bacteriostatic effect during transportation and storage and thus to prevent the formation of endotoxins. To simulate transport conditions, treated and nontreated cotton samples were incubated under high air humidity. The endotoxin contents were determined by Limulus amebocyte lysate assay depending on the duration of incubation. In nontreated samples the endotoxin content grew to over 5,000 ng/mg. In comparison, in treated samples the endotoxin content grew extremely slowly. Thus, the bactericidal treating of raw cotton showed high efficiency as a potential method of byssinosis prevention. The irradiation by gamma-rays is also efficient, but it is not realistic in cotton growing areas of developing countries at the present time. PMID- 12570401 TI - A risk analysis approach to selecting respiratory protection against airborne pathogens used for bioterrorism. AB - The authors present a quantitative risk analysis approach to estimating infection risk due to airborne pathogens exhibiting relatively large infectious dose values. The method is applied to hypothetical scenarios involving airborne spores of Bacillus anthracis. The method combines the estimated parameters for exposure intensity, the pathogen dose-response relationship, and respirator penetration values (if respiratory protection is used). Because knowledge of the true parameter values will be uncertain, an uncertainty analysis is an essential part of the process. Given a specified value for acceptable infection risk, the method permits choosing a respirator that sufficiently reduces exposure to meet the acceptable risk criterion. A strength of the risk analysis approach is its transparency, in that the model structure and data inputs are explicitly identified. Further, risk analysis informs the expert judgment that must typically be applied in selecting respiratory protection against airborne pathogens. PMID- 12570402 TI - Influence of storage on the fungal concentration determination of impinger and filter samples. AB - The effects of storage on concentrations of airborne fungal samples were evaluated in a laboratory test chamber. Spores of Penicillium citrinum and cells of Candida famata var. flareri were aerosolized by a Pitt-3 generator and a Collison nebulizer, respectively. The evaluated sampling methods were AGI-30 impingers, Nuclepore filters, and gelatin filters. The effect of storage time was determined as the ratio, C(t)/C(0), where C(t) and C(0) were the total counts or colony concentrations of the simultaneously collected samples stored for t and 0 hr, respectively. In addition, the effect of storage temperature was investigated by storing AGI-30 and filter samples at 25 and 4 degrees C. The results demonstrated that the culturability of Penicillium spores in the impinger samples decreased as storage time increased. In contrast, the Candida yeast cells could survive and reproduce more cells in the impinger fluid. Moreover, it was observed that refrigeration of the impinger samples could inhibit the growth rate of collected cells. Therefore, it was suggested that the impinger samples should be refrigerated and processed as soon as possible to avoid the loss of spore culturability or the increase of yeast cells. Regarding the filtration methods, the effect of storage time and temperature for Penicillium spores was demonstrated to be insignificant. However, Candida yeast recovery from filters was found to decrease as storage time increased. PMID- 12570403 TI - Effect of building frame and moisture damage on microbiological indoor air quality in school buildings. AB - The effect of building frame and moisture damage on microbial indoor air quality was characterized in 17 wooden and 15 concrete or brick school buildings. Technical investigations to detect visible moisture and mold damage were performed according to a standardized protocol. Viable airborne microbes were determined by using a six-stage impactor (Andersen 10-800). Mean concentrations of viable airborne fungi were significantly higher in wooden schools than in concrete schools, showing that the frame material was a determinant of concentrations of airborne fungi. Moisture damage of the building did not alter the fungal concentrations in wooden school buildings. In contrast, in concrete schools the effect of moisture damage was clearly seen as higher concentrations compared with the reference schools. Aspergillus versicolor, Stachybotrys, and Acremonium were detected only in samples from moisture damaged buildings, and can be considered marker fungi of such damage in school buildings. In addition, the presence of Oidiodendron as well as elevated concentrations of Cladosporium and actinobacteria were associated with moisture damage in concrete schools. PMID- 12570404 TI - Can Chinese MAWL be used for designing manual handling tasks? AB - Previous studies have employed collected MAWL (maximal acceptable weight of lift) data as criteria for designing manual handling tasks because MAWL relates to one's limitation of muscle strength. However, whether the Chinese MAWL can still be used for job design is questionable because the Chinese are relatively weaker than Western subjects in their upper extremities. To clarify this, in the present study 24 male Chinese subjects psychophysically determined their MAWL and MAWLo (maximal acceptable weight of lower) under task conditions of three lifting frequencies and two lifting ranges. The results indicated that subjects' MAWLo were significantly lower than their MAWL, especially when infrequent tasks were performed (including tasks of one time maximum and one lift (lower)/min, all p<.05). This could be attributed to a relatively disadvantageous position in the upper extremities at the beginning stage of these lowering tasks. This finding strongly implies that it is not enough to consider only Chinese MAWL when designing manual handling tasks; MAWLo data should not be ignored. PMID- 12570405 TI - Tunneling measurement of a single quantum spin. AB - Measurement of the tunneling current of spin-polarized electrons via a molecule with a localized spin provides information on the orientation of that spin. We show that a strong tunneling current due to the shot noise suppresses the spin dynamics, such as the spin precession in an external magnetic field, and the relaxation due to the environment (quantum Zeno effect). A weak tunneling current preserves the spin precession with the oscillatory component of the current of the same order as the noise. We propose an experiment to observe the Zeno effect in a tunneling system and describe how the tunneling current may be used to read a qubit represented by a single spin 1/2. PMID- 12570406 TI - Condensate density and superfluid mass density of a dilute Bose-Einstein condensate near the condensation transition. AB - We derive via diagrammatic perturbation theory the scaling behavior of the condensate and superfluid mass density of a dilute Bose gas just below the condensation temperature, T(c). Sufficiently below T(c) particle excitations are described by mean field (Bogoliubov). Near T(c), however, mean field fails, and the system undergoes a second order phase transition, rather than first order as predicted by Bogoliubov theory. Both condensation and superfluidity occur at the same T(c), and have similar scaling functions below T(c), but different finite size scaling at T(c) to leading order in the system size. A self-consistent two loop calculation yields the condensate fraction critical exponent, 2beta approximately 0.66. PMID- 12570407 TI - Dynamically stabilized bright solitons in a two-dimensional bose-einstein condensate. AB - We demonstrate that a matter-wave bright soliton can be stabilized in 2D free space by causing the strength of interactions to oscillate rapidly between repulsive and attractive by using, e.g., Feshbach resonance. PMID- 12570408 TI - Multiscaling of galactic cosmic ray flux. AB - Multiscaling analysis of the differential flux dissipation rate of galactic cosmic rays (carbon nuclei) is performed in the energy ranges 56.3-73.4 MeV/nucleon and 183.1-198.7 MeV/nucleon, using the data collected by the ACE/CRIS spacecraft instrument for the year 2000. The analysis reveals strong (turbulencelike) intermittency of the flux dissipation rate for short-term intervals: 1-30 h. It is also found that the type of intermittency can be different in different energy ranges. PMID- 12570409 TI - Replica limit of the toda lattice equation. AB - In a recent breakthrough Kanzieper showed that it is possible to obtain exact nonperturbative random matrix results from the replica limit of the corresponding Painleve equation. In this article we analyze the replica limit of the Toda lattice equation and obtain exact expressions for the two-point function of the Gaussian unitary ensemble and the resolvent of the chiral unitary ensemble. In the latter case both the fully quenched and the partially quenched limit are considered. This derivation explains in a natural way the appearance of both compact and noncompact integrals, the hallmark of the supersymmetric method, in the replica limit of the expression for the resolvent. PMID- 12570411 TI - n-p short-range correlations from (p,2p+n) measurements. AB - We studied the 12C(p,2p+n) reaction at beam momenta of 5.9, 8.0, and 9.0 GeV/c. For quasielastic (p,2p) events p(f), the momentum of the knocked-out proton before the reaction, was compared (event by event) with p(n), the coincident neutron momentum. For |p(n)|>k(F)=0.220 GeV/c (the Fermi momentum) a strong back to-back directional correlation between p(f) and p(n) was observed, indicative of short-range n-p correlations. From p(n) and p(f) we constructed the distributions of c.m. and relative motion in the longitudinal direction for correlated pairs. We also determined that 49+/-13% of events with |p(f)|>k(F) had directionally correlated neutrons with |p(n)|>k(F). PMID- 12570412 TI - Giant-dipole resonance and the deformation of hot, rotating nuclei. AB - The development of nuclear shapes under the extreme conditions of high angular momentum and/or temperature is examined. Scaling properties are used to demonstrate universal properties of both thermal expectation values of nuclear shapes as well as the minima of the free energy, which can be used to understand the Jacobi transition. A universal correlation between the width of the giant dipole resonance and quadrupole deformation is found, providing a novel probe to measure the nuclear deformation in hot nuclei. PMID- 12570413 TI - Three-body monopole corrections to realistic interactions. AB - It is shown that a very simple three-body monopole term can solve practically all the spectroscopic problems-in the p, sd, and pf shells-that were hitherto assumed to need drastic revisions of the realistic potentials. PMID- 12570410 TI - Indications of neutrino oscillation in a 250 km long-baseline experiment. AB - The K2K experiment observes indications of neutrino oscillation: a reduction of nu(mu) flux together with a distortion of the energy spectrum. Fifty-six beam neutrino events are observed in Super-Kamiokande (SK), 250 km from the neutrino production point, with an expectation of 80.1(+6.2)(-5.4). Twenty-nine one ring mu-like events are used to reconstruct the neutrino energy spectrum, which is better matched to the expected spectrum with neutrino oscillation than without. The probability that the observed flux at SK is explained by statistical fluctuation without neutrino oscillation is less than 1%. PMID- 12570414 TI - Feedback cooling of a one-electron oscillator. AB - A one-electron oscillator is cooled from 5.2 K to 850 mK using electronic feedback. Novel quantum jump thermometry reveals a Boltzmann distribution of oscillator energies and directly measures the corresponding temperature. The ratio of electron temperature and damping rate (also directly measured) is observed to be a fluctuation-dissipation invariant, independent of feedback gain, as predicted for noiseless feedback. The sharply reduced linewidth that results from feedback cooling illustrates the likely importance for improved fundamental measurements and symmetry tests. PMID- 12570415 TI - Nonclassical paths in the recurrence spectrum of diamagnetic atoms. AB - Using time-independent scattering matrices, we study how the effects of nonclassical paths on the recurrence spectra of diamagnetic atoms can be extracted from purely quantal calculations. This study reveals an intimate relationship between two types of nonclassical paths: exotic ghost orbits and diffractive orbits. This relationship proves to be a previously unrecognized reason for the success of semiclassical theories, such as closed-orbit theory, and permits a comprehensive reformulation of the semiclassical theory that elucidates its convergence properties. PMID- 12570416 TI - Direct versus sequential double ionization of Mg with extreme-ultraviolet radiation. AB - We present an ab initio calculation of multiphoton double ionization in atomic magnesium, the first of its type in an atom with a complex core, showing that direct correlated double electron ejection can be studied through analysis of the photoelectron energy spectrum, using currently or upcoming extreme-ultraviolet radiation sources. PMID- 12570417 TI - Simple iterative construction of the optimized effective potential for orbital functionals, including exact exchange. AB - For exchange-correlation functionals that depend explicitly on the Kohn-Sham orbitals, the potential V(xcsigma)(r) must be obtained as the solution of the optimized effective potential (OEP) integral equation. This is very demanding and has limited the use of orbital functionals. We demonstrate that instead the OEP can be obtained iteratively by solving the partial differential equations for the orbital shifts that exactify the Krieger-Li-Iafrate approximation. Unoccupied orbitals do not need to be calculated. Accuracy and efficiency of the method are shown for atoms and clusters using the exact-exchange energy. Counterintuitive asymptotic limits of the exact OEP are presented. PMID- 12570418 TI - Excitations in time-dependent density-functional theory. AB - An approximate solution to the time-dependent density-functional theory response equations for finite systems is developed, yielding corrections to the single pole approximation. These explain why allowed Kohn-Sham transition frequencies and oscillator strengths are usually good approximations to the true values, and why sometimes they are not. The approximation yields simple expressions for Gorling-Levy perturbation theory results, and a method for estimating expectation values of the unknown exchange-correlation kernel. PMID- 12570419 TI - Linking ultracold polar molecules. AB - We predict that pairs of polar molecules can be weakly bound together in an ultracold environment, provided that a dc electric field is present. The field that links the molecules together also strongly influences the basic properties of the resulting dimer, such as its binding energy and predissociation lifetime. Because of their long-range character, these dimers will be useful in disentangling cold collision dynamics of polar molecules. As an example, we estimate the microwave photoassociation yield for OH-OH cold collisions. PMID- 12570420 TI - Discovery of temperature-dependent phenomena of muon-catalyzed fusion in solid deuterium and tritium mixtures. AB - A systematic experimental study on muon-catalyzed fusion was conducted using a series of solid deuterium and tritium mixtures. A variety of conditions were investigated, i.e., tritium concentrations from 20% to 70%, and temperatures from 5 to 16 K. With decreasing temperature, we observed an unexpected decrease in the muon cycling rate (lambda(c)) and an increase in the muon loss probability (W). The origins of these observed changes were interpreted by the temperature dependence in the dt mu formation process for lambda(c) and that in the muon reactivation process after muon-to-alpha sticking for W. PMID- 12570421 TI - Experimental investigation of criteria for continuous variable entanglement. AB - We generate a pair of entangled beams from the interference of two amplitude squeezed beams. The entanglement is quantified in terms of EPR paradox and inseparability criteria, with both results clearly beating the standard quantum limit. We experimentally analyze the effect of decoherence on each criterion and demonstrate qualitative differences. We also characterize the number of required and excess photons present in the entangled beams and provide contour plots of the efficacy of quantum information protocols in terms of these variables. PMID- 12570422 TI - Nonlocal pulse shaping with entangled photon pairs. AB - Nonlocal shaping effects in the time or spectral profiles of an entangled photon pair emerging from a pulsed parametric down-converter are observed by spectrally or temporally filtering one of the twin beams. In particular, we demonstrate the appearance of fourth-order ("ghost") interference fringes in the spectrum of one beam conditioned by photodetection at the output of an unbalanced Michelson interferometer placed in the path of the other beam. The coherence time of the pump is the limiting factor for the sharpness of the details in the shaped biphoton spectrum. PMID- 12570423 TI - Rayleigh scattering and atomic dynamics in dissipative optical lattices. AB - We investigate Rayleigh scattering in dissipative optical lattices. In particular, following recent proposals [S. Guibal, Phys. Rev. Lett. 78, 4709 (1997)]; C. Jurczak, Phys. Rev. Lett. 77, 1727 (1996)]], we study whether the Rayleigh resonance originates from the diffraction on a density grating and is therefore a probe of transport of atoms in optical lattices. It turns out that this is not the case: the Rayleigh line is instead a measure of the cooling rate, while spatial diffusion contributes to the scattering spectrum with a much broader resonance. PMID- 12570424 TI - Resonant second-order nonlinear optical processes in quantum cascade lasers. AB - We demonstrate an efficient intracavity nonlinear interaction of laser modes in a specially adapted quantum cascade laser. A two-wavelength quantum cascade laser structure emitting at wavelengths of 7.1 and 9.5 micrometer included cascaded resonant optical intersubband transitions in an intracavity configuration leading to resonantly enhanced sum-frequency and second-harmonic generation at wavelengths of 4.1, 3.6, and 4.7 micrometer respectively. Laser peak optical powers of 60 and 80 mW resulted in 30 nW of sum-frequency signal and 10-15 nW of second-harmonic signal, both in good agreement with theoretical calculations. PMID- 12570425 TI - Quantum chaos triggered by precursors of a quantum phase transition: the dicke model. AB - We consider the Dicke Hamiltonian, a simple quantum-optical model which exhibits a zero-temperature quantum phase transition. We present numerical results demonstrating that at this transition the system changes from being quasi integrable to quantum chaotic. By deriving an exact solution in the thermodynamic limit we relate this phenomenon to a localization-delocalization transition in which a macroscopic superposition is generated. We also describe the classical analogs of this behavior. PMID- 12570426 TI - Observation of locked intrinsic localized vibrational modes in a micromechanical oscillator array. AB - The nonlinear vibrational properties of a periodic micromechanical oscillator array have been measured. For sufficiently large amplitude of the driver, the optic mode of the di-element cantilever array becomes unstable and breaks up into excitations ranging over only a few cells. A driver-induced locking effect is observed to eternalize some of these intrinsic localized modes so that their amplitudes become fixed and the modes become spatially pinned. PMID- 12570427 TI - Granular avalanches in fluids. AB - Three regimes of granular avalanches in fluids are put in light depending on the Stokes number St which prescribes the relative importance of grain inertia and fluid viscous effects and on the grain/fluid density ratio r. In gas (r>>1 and St>1, e.g., the dry case), the amplitude and time duration of avalanches do not depend on any fluid effect. In liquids (r approximately 1), for decreasing St, the amplitude decreases and the time duration increases, exploring an inertial regime and a viscous regime. These three regimes are described by the analysis of the elementary motion of one grain. PMID- 12570428 TI - Magnetic propulsion of intense lithium streams in a tokamak magnetic field. AB - This paper describes the effect and gives the theory of magnetic propulsion which allows driving free surface plasma facing liquid lithium streams in tokamaks. In the approximation of a thin flowing layer the MHD equations are reduced to one integrodifferential equation which takes into account the propulsion effect, viscosity, and the drag force due to magnetic pumping and other interactions with the magnetic field. A stability criterion is obtained for stabilization of the "sausage" instability of the streams by centrifugal force. PMID- 12570429 TI - Optical manipulation of defects in a lyotropic lamellar phase. AB - Here we study the line defect in a hyperswollen lamellar phase of lyotropic liquid crystal by applying a laser trapping method. We have succeeded in directly measuring the tension of a single isolated line defect and the adhesion energy between two defects. We demonstrate a new possibility of intentional patterning of various defects by direct optical manipulation. Furthermore, local rheological measurements provide information on the membrane organization around a particle and also evidence suggesting that flow in a lamellar phase has a two-dimensional nature. PMID- 12570431 TI - Cooperative Jahn-Teller phase transition in LaMnO3 studied by X-ray absorption spectroscopy. AB - The local structure of LaMnO3 across the Jahn-Teller (JT) transition at T(JT)=750 K was studied by means of x-ray absorption near edge structure and extended x-ray absorption fine structure at the Mn K-edge. Our results indicate a similar electronic local structure for Mn atoms above and below T(JT) and a dynamical tetragonal JT distortion of MnO6 octahedra above T(JT). The structural transition is originated by the ordering of tetragonally distorted octahedra. The entropy content of the transition is analyzed within the framework of the three-state Potts model with nearest neighbor antiferrodistortive coupling. PMID- 12570430 TI - Adaptive crystal structures: CuAu and NiPt. AB - We discover that Au-rich Cu1-xAux and Pt-rich Ni1-xPtx contain a composition range in which there is a quasicontinuum of stable, ordered "adaptive structures" made of (001) repeat units of simple structural motifs. This is found by searching approximately 3x10(6) different fcc configurations whose energies are parametrized via a "cluster expansion" of first-principles-calculated total energies of just a few structures. This structural adaptivity is explained in terms of an anisotropic, long-range strain energy. PMID- 12570432 TI - Origin of the universal roughness exponent of brittle fracture surfaces:stress weighted percolation in the damage zone. AB - We suggest that the observed large-scale universal roughness of brittle fracture surfaces is due to the fracture propagation being a damage coalescence process described by a stress-weighted percolation phenomenon in a self-generated quadratic damage gradient. We use the quasistatic 2D fuse model as a paradigm of a mode I fracture model. We measure for this model, which exhibits a correlated percolation process, the correlation length exponent nu approximately 1.35 and conjecture it to be equal to that of classical percolation, 4/3. We then show that the roughness exponent in the 2D fuse model is zeta=2nu/(1+2nu)=8/11. This is in accordance with the numerical value zeta=0.75. Using the value for 3D percolation, nu=0.88, we predict the roughness exponent in the 3D fuse model to be zeta=0.64, in close agreement with the previously published value of 0.62+/ 0.05. We furthermore predict zeta=4/5 for 3D brittle fractures, based on a recent calculation giving nu=2. This is in full accordance with the value zeta=0.80 found experimentally. PMID- 12570433 TI - Roughness of interfacial crack fronts: stress-weighted percolation in the damage zone. AB - We study numerically the roughness exponent zeta of an in-plane fracture front slowly propagating along a heterogeneous interface embedded in an elastic body, using a model based on the evolution of a process zone rather than a fracture line. We find zeta=0.60+/-0.05. For the first time, simulation results are in close agreement with experimental results. We then show that the roughness exponent is related to the correlation length exponent nu of a stress-weighted percolation problem through zeta=nu/(1+nu). A numerical study of the stress weighted percolation problem yields nu=1.54 giving zeta=0.61 in close agreement with our numerical results and with experimental observations. PMID- 12570434 TI - Dynamics of large-scale plastic deformation and the necking instability in amorphous solids. AB - We use the shear transformation zone (STZ) theory of dynamic plasticity to study the necking instability in a two-dimensional strip of amorphous solid. Our Eulerian description of large-scale deformation allows us to follow the instability far into the nonlinear regime. We find a strong rate dependence; the higher the applied strain rate, the further the strip extends before the onset of instability. The material hardens outside the necking region, but the description of plastic flow within the neck is distinctly different from that of conventional time-independent theories of plasticity. PMID- 12570435 TI - Fluid adsorption near an apex: covariance between complete and critical wetting. AB - Critical wetting is an elusive phenomenon for solid-fluid interfaces. Using interfacial models we show that the diverging length scales, which characterize complete wetting at an apex, precisely mimic critical wetting with the apex angle behaving as the contact angle. Transfer matrix, renormalization group, and mean field analysis show that this covariance is obeyed in 2D and 3D and for long- and short-ranged forces. This connection should be experimentally accessible and provides a means of checking theoretical predictions for critical wetting. PMID- 12570436 TI - Atomistics of Ge deposition on Si(100) by atomic layer epitaxy. AB - Chlorine termination of mixed Ge/Si(100) surfaces substantially enhances the contrast between Ge and Si sites in scanning tunneling microscopy observations. This finding enables a detailed investigation of the spatial distribution of Ge atoms deposited on Si(100) by atomic layer epitaxy. The results are corroborated by photoemission measurements aided by an unusually large chemical shift between Cl adsorbed on Si and Ge. Adsorbate-substrate atomic exchange during growth is shown to be important. The resulting interface is thus graded, but characterized by a very short length scale of about one monolayer. PMID- 12570437 TI - First-principles atomistic thermodynamics for oxidation catalysis: surface phase diagrams and catalytically interesting regions. AB - Present knowledge of the function of materials is largely based on studies (experimental and theoretical) that are performed at low temperatures and ultralow pressures. However, the majority of everyday applications, like, e.g., catalysis, operate at atmospheric pressures and temperatures at or higher than 300 K. Here we employ ab initio, atomistic thermodynamics to construct a phase diagram of surface structures in the (T,p) space from ultrahigh vacuum to technically relevant pressures and temperatures. We emphasize the value of such phase diagrams as well as the importance of the reaction kinetics that may be crucial, e.g., close to phase boundaries. PMID- 12570438 TI - Role of bulk thermal defects in the reconstruction dynamics of the TiO2(110) surface. AB - We use low-energy electron microscopy to show that changing the temperature of oxygen-deficient, rutile-structure crystals causes steps on the (110) surfaces to move. This motion occurs because the concentration of bulk oxygen vacancies changes with temperature, requiring that material be added to or subtracted from the surface. During cooling below a bulk-stoichiometry-dependent temperature, the surface reconstructs into a 1x2 structure in the regions surface steps have swept through, showing that the structural and compositional changes needed to form the 1x2 phase are facilitated by the surface-to-bulk mass flow. PMID- 12570439 TI - Novel optical and magnetic bistability and photoinduced transition in a one dimensional halogen-bridged binuclear pt complex. AB - In iodine-bridged binuclear Pt compounds, R(4)[Pt(2)(pop)(4)I]nH(2)O and R(')(2)[Pt(2)(pop)(4)I]nH(2)O (pop=P(2)O5H2-2), electronic structures on the PtPtI chains have been controlled between a diamagnetic charge-density-wave (CDW) state and a paramagnetic charge-polarization (CP) state by modification of the counterions (R, R') located between chains. In the R=(C2H5)2NH+2 compound, a pressure-induced CP to CDW transition with a drastic color change is identified. This transition is accompanied by a large hysteresis loop within which photoinduced transition between CDW and CP can be driven by selecting the excitation photon energy. PMID- 12570440 TI - Nonlinear screening in two-dimensional electron gases. AB - We have performed self-consistent calculations of the nonlinear screening of a point charge Z in a two-dimensional electron gas using a density functional theory method. We find that the screened potential for a Z=1 charge supports a bound state even in the high-density limit where one might expect perturbation theory to apply. To explain this behavior, we prove a theorem to show that the results of linear response theory are in fact correct even though bound states exist. PMID- 12570441 TI - Charge transport transitions and scaling in disordered arrays of metallic dots. AB - We examine the charge transport through disordered arrays of metallic dots using numerical simulations. We find power law scaling in the current-voltage curves for arrays containing no voids, while for void-filled arrays charge bottlenecks form and a single scaling is absent, in agreement with recent experiments. In the void-free case we also show that the scaling exponent depends on the effective dimensionality of the system. For increasing applied drives we find a transition from 2D disordered filamentary flow near threshold to a 1D smectic flow which can be identified experimentally using characteristics in the transport curves and conduction noise. PMID- 12570442 TI - Two-electron photon emission from metallic quantum wells. AB - Unusual emission of visible light is observed in scanning tunneling microscopy of the quantum well system Na on Cu(111). Photons are emitted at energies exceeding the energy of the tunneling electrons. Model calculations of two-electron processes which lead to quantum well transitions reproduce the experimental fluorescence spectra, the quantum yield, and the power-law variation of the intensity with the excitation current. PMID- 12570443 TI - Linear and nonlinear photoexcitation dynamics in pi-conjugated polymers. AB - Linear and nonlinear recombination kinetics with various lifetime distributions were identified for long-lived photoexcitations in a series of pi-conjugated polymer films using modulation frequency and excitation intensity dependencies of the photoinduced absorption. This includes monomolecular, bimolecular, and defect limited recombination processes that lead to saturation. Using generalized kinetics parameters, we found characteristic plots for all recombination processes. Specifically, the bimolecular recombination process shows superlinear intensity dependence away from the steady state; on the contrary, dispersive bimolecular recombination leads to sublinear dependence. PMID- 12570444 TI - Nonlinear quasiparticle tunneling between fractional quantum hall edges. AB - Remarkable nonlinearities in the differential tunneling conductance between fractional quantum Hall edge states at a constriction are observed in the weak backscattering regime. In the nu=1/3 state a peak develops as temperature is increased and its width is determined by the fractional charge. In the range 2/3J(t), the electric field penetrating from current leads gives rise to various static and dynamic soliton phase textures, and voltage oscillations similar to the nonstationary Josephson effect. We propose experiments to observe these effects which would probe the multicomponent nature of the superconducting order parameter. PMID- 12570452 TI - Domain wall depinning in random media by ac fields. AB - The viscous motion of an interface driven by an ac external field of frequency omega(0) in a random medium is considered here in the nonadiabatic regime. The velocity exhibits a smeared depinning transition showing a double hysteresis which is absent in the adiabatic case omega(0)-->0. Using scaling arguments and an approximate renormalization group calculation we explain the main characteristics of the hysteresis loop. In the low frequency limit these can be expressed in terms of the depinning threshold and the critical exponents of the adiabatic case. PMID- 12570453 TI - Anomalous Hall effect as a probe of the chiral order in spin glasses. AB - The anomalous Hall effect arising from the noncoplanar spin configuration (chirality) is discussed as a probe of the chiral order in spin glasses. It is shown that the Hall coefficient yields direct information about the linear and nonlinear chiral susceptibilities of the spin sector, which has been hard to obtain experimentally from the standard magnetic measurements. Based on the chirality scenario of spin-glass transition, predictions are given on the behavior of the Hall resistivity of canonical spin glasses. PMID- 12570454 TI - Algorithm for linear response functions at finite temperatures: application to ESR spectrum of s=1/2 antiferromagnet Cu benzoate. AB - We introduce an efficient and numerically stable method for calculating linear response functions chi(q,omega) of quantum systems at finite temperatures. The method is a combination of numerical solution of the time-dependent Schrodinger equation, random vector representation of trace, and Chebyshev polynomial expansion of Boltzmann operator. This method should be very useful for a wide range of strongly correlated quantum systems at finite temperatures. We present an application to the ESR spectrum of s=1 / 2 antiferromagnet Cu benzoate. PMID- 12570455 TI - Ruderman-Kittel-Kasuya-Yosida-like ferromagnetism in MnxGe1-x. AB - The nature and origin of ferromagnetism in magnetic semiconductors is investigated by means of highly precise electronic and magnetic property calculations on MnxGe1-x as a function of the location of Mn sites in a large supercell. Surprisingly, the coupling is not always ferromagnetic (FM), even for large Mn-Mn distances. The exchange interaction between Mn ions oscillates as a function of the distance between them and obeys the Ruderman-Kittel-Kasuya-Yosida analytic formula. The estimated Curie temperature is in good agreement with recent experiments, and the estimated effective magnetic moment is about 1.7mu(B)/Mn, in excellent agreement with the experimental values, (1.4 1.9)mu(B)/Mn. PMID- 12570456 TI - Rigorous decimation-based construction of ground pure states for spin-glass models on random lattices. AB - A constructive scheme for determining pure states at very low temperature in the 3-spins glass model on a random lattice is provided, in full agreement with Parisi's one step replica symmetry breaking (RSB) scheme. Proof is based on the analysis of a partial decimation procedure and of the statistical properties of its output, i.e., a reduced Hamiltonian acting on a subset of the initial spins. The number of ground states (GS) in each state, the number of states, and the distances between GS are calculated and correspond to RSB predictions. PMID- 12570457 TI - Obtaining absorptive line shapes in two-dimensional infrared vibrational correlation spectra. AB - Absorptive line shapes in two-dimensional infrared (2D IR) vibrational spectra are important for an intuitive interpretation of molecular structure and dynamics. We obtain an absorptive 2D IR correlation spectrum by summing complementary spectra from experiments sampling vibrational coherences that oscillate with conjugate frequencies in the initial evolution time period. The 2D correlation spectrum of a coupled vibrational system reveals certain spectral features with tilted line shapes that are explained in terms of unequal contributions of Liouville-space pathways. PMID- 12570458 TI - Time domain investigation of the intrachain vibrational dynamics of a prototypical light-emitting conjugated polymer. AB - Coherent nuclear motion of the carbon backbone is monitored in real time in substituted poly(phenylene)-vinylene in both excited and ground electronic state using sub-10-fs light pulses. Characteristic features of the intrachain vibrational dynamics are obtained. We observe vibrational dephasing in the excited state within 1 ps (time constant T(2e) approximately 300 fs). These findings are in agreement with the molecular picture for photoexcitation in conjugated polymers. PMID- 12570459 TI - Observing photochemical transients by ultrafast x-ray absorption spectroscopy. AB - Accurate determination of the transient electronic structures, which drive photochemical reactions, is crucial in chemistry and biology. We report the detection of transient chemical changes on the picosecond time scale by x-ray absorption near-edge structure of photoexcited aqueous [Ru(bpy)(3)](2+). Upon ultrashort laser pulse excitation a charge transfer excited state having a 300 ns lifetime is formed. We detect the change of oxidation state of the central Ru atom at its L3 and L2 edges, at a temporal resolution of 100 ps with the zero of time unambiguously determined. PMID- 12570460 TI - Quantum computing with spin cluster qubits. AB - We study the low energy states of finite spin chains with isotropic (Heisenberg) and anisotropic (XY and Ising-like) antiferromagnetic exchange interaction with uniform and nonuniform coupling constants. We show that for an odd number of sites a spin cluster qubit can be defined in terms of the ground state doublet. This qubit is remarkably insensitive to the placement and coupling anisotropy of spins within the cluster. One- and two-qubit quantum gates can be generated by magnetic fields and intercluster exchange, and leakage during quantum gate operation is small. Spin cluster qubits inherit the long decoherence times and short gate operation times of single spins. Control of single spins is hence not necessary for the realization of universal quantum gates. PMID- 12570461 TI - Local indistinguishability: more nonlocality with less entanglement. AB - We provide a first operational method for checking local indistinguishability of orthogonal states. It originates from that in Ghosh et al. [Phys. Rev. Lett. 87, 5807 (2001)]], though we deal with pure states. Our method shows that probabilistic local distinguishing is possible for a complete multipartite orthogonal basis if and only if all vectors are product. Also, it leads to local indistinguishability of a set of orthogonal pure states of 3 multiply sign in circle 3, which shows that one can have more nonlocality with less entanglement, where "more nonlocality" is in the sense of "increased local indistinguishability of orthogonal states." This is, to our knowledge, the only known example where d orthogonal states in d multiply sign in circle d are locally indistinguishable. PMID- 12570462 TI - Using distributed nonlinear dynamics for public key encryption. AB - We introduce a new method for asymmetric (public key/private key) encryption exploiting properties of nonlinear dynamical systems. A high-dimensional dissipative nonlinear dynamical system is distributed between transmitter and receiver, so we call the method distributed dynamics encryption (DDE). The transmitter dynamics is public, and the receiver dynamics is hidden. A message is encoded by modulation of parameters of the transmitter, and this results in a shift of the overall system attractor. An unauthorized receiver does not know the hidden dynamics in the receiver and cannot decode the message. We present an example of DDE using a coupled map lattice. PMID- 12570463 TI - Entangling power of passive optical elements. AB - We investigate the entangling capability of passive optical elements, both qualitatively and quantitatively. We present a general necessary and sufficient condition for the possibility of creating distillable entanglement in an arbitrary multimode Gaussian state with the help of passive optical elements, thereby establishing a general connection between squeezing and the entanglement that is attainable by nonsqueezing operations. Special attention is devoted to general two-mode Gaussian states, for which we provide the optimal entangling procedure, present an explicit formula for the attainable degree of entanglement, and discuss several practically important special cases. PMID- 12570464 TI - Entanglement and entropy engineering of atomic two-qubit States. AB - We propose a scheme employing quantum-reservoir engineering to controllably entangle the internal states of two atoms trapped in a high-finesse optical cavity. Using laser and cavity fields to drive two separate Raman transitions between stable atomic ground states, a system is realized corresponding to a pair of two-state atoms coupled collectively to a squeezed reservoir. Phase-sensitive reservoir correlations lead to entanglement between the atoms, and, via local unitary transformations and adjustment of the degree and purity of squeezing, one can prepare entangled mixed states with any allowed combination of linear entropy and entanglement of formation. PMID- 12570465 TI - Direct measurement of entropic forces induced by rigid rods. AB - We present the first direct depletion potential measurements for a single colloidal sphere close to a wall in a suspension of rigid colloidal rods. Since all particle interactions are as good as hard-core-like, the depletion potentials are of entirely entropic origin. We developed a density functional theory approach that accurately accounts for this experimental situation. The depletion potentials calculated for different rod number densities are in very good quantitative agreement with the experimental results. PMID- 12570466 TI - Stability of soap films: hysteresis and nucleation of black films. AB - We study the stability of soap films of a nonionic surfactant under different applied capillary pressures on the film. Depending on the pressure, either a thick common black film (CBF), or a micro-scopically thin Newton black film (NBF) is formed as a (metastable) equilibrium state, with a first-order (discontinuous) transition between the two. Studying the dynamics of the CBF-NBF transition, it is found that under certain conditions a hysteresis for the transition is observed: for a given range of pressures, either of the two states may be observed. We quantify the nucleation process that is at the basis of these observations both experimentally and theoretically. PMID- 12570467 TI - Monte carlo study of coulombic criticality in polyelectrolytes. AB - The role of charges in determining the water solubility of polyelectrolytes, a question of considerable relevance to biology, is currently unresolved. We use computer simulations to study the purely Coulombic phase separation of flexible polyelectrolytes with monovalent counterions in an athermal solvent. In agreement with recent theories we find that the critical temperature for this transition increases with chain length, but that the critical density remains unchanged. We therefore stress that the phase behavior of polyelectrolytes is qualitatively different from uncharged polymers, where the critical density decreases towards zero for long chains. PMID- 12570468 TI - Phase behavior of thermally responsive microgel colloids. AB - The phase behavior of poly-N-isopropylacrylamide (PNIPAM) nanoparticles dispersed in water is investigated using a thermodynamic perturbation theory combined with light-scattering and spectrometer measurements. It is shown how the volume transition of PNIPAM particles affects the interaction potential and determines a novel phase diagram that has not been observed in conventional colloids. Because both particle size and attractive potential depend on temperature, PNIPAM aqueous dispersion exhibits phase transitions at a fixed particle number density by either increasing or decreasing temperature. PMID- 12570469 TI - Comment on "Microscopic mechanisms of electric-field-induced alignment of block copolymer microdomains". PMID- 12570471 TI - Comment on "Orthorhombic intermediate state in the zinc blende to rocksalt transformation path of SiC at high pressure". PMID- 12570473 TI - Condensate statistics in one-dimensional interacting Bose gases: exact results. AB - Recently, a quantum Monte Carlo method alternative to the path integral Monte Carlo method was developed for solving the N-boson problem; it is based on the stochastic evolution of classical fields. Here we apply it to obtain exact results for the occupation statistics of the condensate mode in a weakly interacting trapped one-dimensional Bose gas. The temperature is varied across the critical region down to temperatures lower than the trap level spacing. We also derive the condensate statistics in the Bogoliubov theory: this reproduces the exact results at low temperature and explains the suppression of odd numbers of noncondensed particles at T approximately 0. PMID- 12570474 TI - Creating maximally entangled atomic states in a Bose-Einstein condensate. AB - We propose a protocol to create maximally entangled pairs, triplets, quartiles, and other clusters of Bose-condensed atoms starting from a condensate in the Mott insulator state. The essential element is to drive single atom Raman transitions using laser pulses. Our scheme is simple, efficient, and can be readily applied to the recent experimental system as reported by M. Greiner 413, 44 (2002)]. PMID- 12570475 TI - Coherent collisions between Bose-Einstein condensates. AB - We study the nondegenerate parametric amplifier for matter waves, implemented by colliding two Bose-Einstein condensates. The coherence of the amplified waves is shown by observing high contrast interference with a reference wave and by reversing the amplification process. Since our experiments also place limits on all known sources of decoherence, we infer that relative number squeezing is most likely present between the amplified modes. Finally, we suggest that reversal of the amplification process may be used to detect relative number squeezing without requiring subshot-noise detection. PMID- 12570476 TI - Cooling dynamics of ultracold two-species Fermi-Bose mixtures. AB - We compare strategies for evaporative and sympathetic cooling of two-species Fermi-Bose mixtures in single-color and two-color optical dipole traps. We show that in the latter case a large heat capacity of the bosonic species can be maintained during the entire cooling process. This could allow one to efficiently achieve a deep Fermi degeneracy regime having at the same time a significant thermal fraction for the Bose gas, crucial for a precise thermometry of the mixture. Two possible signatures of a superfluid phase transition for the Fermi species are discussed. PMID- 12570477 TI - Doubly stochastic coherence via noise-induced symmetry in bistable neural models. AB - The generation of coherent dynamics due to noise in an activator-inhibitor system describing bistable neural dynamics is investigated. We show that coherence can be induced in deterministically asymmetric regimes via symmetry restoration by multiplicative noise, together with the action of additive noise which induces jumps between the two stable steady states. The phenomenon is thus doubly stochastic, because both noise sources are necessary. This effect can be understood analytically in the frame of a small-noise expansion and is confirmed experimentally in a nonlinear electronic circuit. Finally, we show that spatial coupling enhances this coherent behavior in a form of system-size coherence resonance. PMID- 12570478 TI - Cosmological parameters are dressed. AB - In the context of the averaging problem in relativistic cosmology, we provide a key to the interpretation of cosmological parameters by taking into account the actual inhomogeneous geometry of the Universe. We discuss the relation between "bare" cosmological parameters determining the cosmological model and the parameters interpreted by observers with a "Friedmannian bias," which are "dressed" by the smoothed-out geometrical inhomogeneities of the surveyed spatial region. PMID- 12570479 TI - Parametrization of dark-energy properties: a principal-component approach. AB - Considerable work has been devoted to the question of how best to parametrize the properties of dark energy, in particular, its equation of state w. We argue that, in the absence of a compelling model for dark energy, the parametrizations of functions about which we have no prior knowledge, such as w(z), should be determined by the data rather than by our ingrained beliefs or familiar series expansions. We find the complete basis of orthonormal eigenfunctions in which the principal components [weights of w(z)] that are determined most accurately are separated from those determined most poorly. Furthermore, we show that keeping a few of the best-measured modes can be an effective way of obtaining information about w(z). PMID- 12570481 TI - Domination of black hole accretion in brane cosmology. AB - We consider the evolution of primordial black holes formed during the high energy phase of the braneworld scenario. We show that the effect of accretion from the surrounding radiation bath is dominant compared to evaporation for such black holes. This feature lasts till the onset of matter (or black hole) domination of the total energy density which could occur either in the high energy phase or later. We find that the black hole evaporation times could be significantly large even for black holes with small initial mass to survive until several cosmologically interesting eras. PMID- 12570480 TI - Quantum cosmology and conformal invariance. AB - According to Belinsky, Khalatnikov, and Lifshitz, gravity near a spacelike singularity reduces to a set of decoupled one-dimensional mechanical models at each point in space. We point out that these models fall into a class of conformal mechanical models first introduced by de Alfaro, Fubini, and Furlan (DFF). The deformation used by DFF to render the spectrum discrete corresponds to a negative cosmological constant. The wave function of the Universe is the zero energy eigenmode of the Hamiltonian, or the spherical vector of the representation of the conformal group SO(1,2). A new class of conformal quantum mechanical models with enhanced ADE symmetry is constructed, based on the quantization of nilpotent coadjoint orbits. PMID- 12570482 TI - Black hole evaporation with separated fermions. AB - In models with a low quantum gravity scale, fast proton decay can be avoided by localizing quarks and leptons to separated positions in an extra 1/TeV sized dimension with gauge and Higgs fields living throughout. Black holes with masses of the order of the quantum gravity scale are therefore expected to evaporate nonuniversally, preferentially radiating directly into quarks or leptons but not both. Should black holes be copiously produced at a future hadron collider, we find the ratio of final state jets to charged leptons to photons is 113:8:1, which differs from previous analyses that assumed all standard model fields live at the same point in the extra dimensional space. PMID- 12570483 TI - Resummed photon spectrum in radiative upsilon decays. AB - We present a theoretical prediction for the photon spectrum in radiative upsilon decay including the effects of resumming the end point region, E(gamma)- >M(upsilon)/2. Our approach is based on nonrelativistic QCD (NRQCD) and the soft collinear effective theory. We find that our results give much better agreement with data than the leading order NRQCD prediction. PMID- 12570485 TI - Two-step enantio-selective optical switch. AB - We present an optical "enantio-selective switch" that, in two steps, turns a ("racemic") mixture of left-handed and right-handed chiral molecules into the enantiomerically pure state of interest. The optical switch is composed of an "enantio-discriminator" and an "enantio-converter" acting in tandem. The method is robust, insensitive to decay processes, and does not require molecular preorientation. We demonstrate the method on the purification of a racemate of (transiently chiral) D2S2 molecules, performed on the nanosecond time scale. PMID- 12570486 TI - Observation of a transition in the dynamics of strong-field double ionization. AB - The double ionization of argon and xenon in an intense laser field has been studied in detail using an electron-ion coincidence technique. The observed double ionization electron spectra in xenon show resonancelike structures here resolved for the first time. In argon, the featureless spectra are consistent with rescattering. This represents a clear transition in the dynamics of strong field double ionization, analogous to the well-known transition between the tunneling and multiphoton regimes in single ionization. PMID- 12570487 TI - Sensitive detection of cold cesium molecules formed on Feshbach resonances. AB - We observe the dynamic formation of quasibound Cs2 molecules near Feshbach resonances in a cold sample of atomic cesium. Using an external probe beam, more than 15 weakly coupled molecular states are detected with high sensitivity, whose collisional formation cross sections are as small as sigma=2 x 10(-16) cm(2). By modeling the molecule formation and dissociation processes with rate equations, we conclude that at an atomic density of 10(13) cm(-3) and temperature of 5 microK, more than 5(1)x10(5) Cs2 molecules in a single rovibrational state coexist with 10(8) Cs atoms in our trap. PMID- 12570488 TI - Element- and size-dependent electron delocalization in AuNX+ clusters (X = Sc, Ti, V, Cr, Mn, Fe, Co, Ni). AB - We investigated the stability of gold clusters doped with open 3d-shell atoms (Sc, Ti, V, Cr, Mn, Fe, Co, Ni). Steps, peaks, and odd-even staggering in mass abundance spectra upon photofragmentation provide evidence for enhanced stability for specific cluster sizes. The observed magic numbers are explained in terms of size- and dopant-dependent modifications of the effective mean-field potential within a phenomenological shell-model approach. Element-dependent 3d electron delocalization and odd-even staggering amplitudes are related to the dopant-atom structure. PMID- 12570489 TI - Observation of a chiral state in a microwave cavity. AB - A microwave experiment has been realized to measure the phase difference of the oscillating electric field at two points inside the cavity. The technique has been applied to a dissipative resonator which exhibits a singularity-called exceptional point-in its eigenvalue and eigenvector spectrum. At the singularity, two modes coalesce with a phase difference of pi/2. We conclude that the state excited at the singularity has a definitive chirality. PMID- 12570490 TI - Arithmetical method to detect integrability in maps. AB - We develop a method to detect the presence of integrals of the motion in symplectic rational maps, by representing these maps over finite fields and examining their orbit structure. We find markedly different orbit statistics depending upon whether the map is integrable or not. PMID- 12570491 TI - Decay of scalar turbulence revisited. AB - We demonstrate that at long times the rate of passive scalar decay in a turbulent, or simply chaotic, flow is dominated by regions where mixing is less efficient. We examine two situations. The first is of a spatially homogeneous stationary turbulent flow with both viscous and inertial scales present. It is shown that at large times scalar fluctuations decay algebraically in time at all spatial scales. The second example explains chaotic stationary flow in a disk/pipe. The boundary region of the flow controls the long-time decay, which is algebraic at some transient times, but becomes exponential, with the decay rate dependent on the scalar diffusion coefficient, at longer times. PMID- 12570484 TI - Azimuthal anisotropy and correlations in the hard scattering regime at RHIC. AB - Azimuthal anisotropy (v(2)) and two-particle angular correlations of high p(T) charged hadrons have been measured in Au+Au collisions at sqrt[s(NN)]=130 GeV for transverse momenta up to 6 GeV/c, where hard processes are expected to contribute significantly. The two-particle angular correlations exhibit elliptic flow and a structure suggestive of fragmentation of high p(T) partons. The monotonic rise of v(2)(p(T)) for p(T)<2 GeV/c is consistent with collective hydrodynamical flow calculations. At p(T)>3 GeV/c, a saturation of v(2) is observed which persists up to p(T)=6 GeV/c. PMID- 12570492 TI - Ultimate state of thermal convection. AB - The ultimate regime of thermal convection, the so-called Kraichnan regime [R. H. Kraichnan, Phys. Fluids 5, 1374 (1962)]], hitherto has been elusive. Here numerical evidence for that regime is presented by performing simulations of the bulk of turbulence only, eliminating the thermal and kinetic boundary layers and replacing them with periodic boundary conditions. PMID- 12570493 TI - Experimental study for the feasibility of a crystalline undulator. AB - We present an idea for creation of a crystalline undulator and report its first realization. One face of a silicon crystal was given periodic microscratches (grooves) by means of a diamond blade. The x-ray tests of the crystal deformation due to a given periodic pattern of surface scratches have shown that a sinusoidal like shape is observed on both the scratched surface and the opposite (unscratched) face of the crystal; that is, a periodic sinusoidal-like deformation goes through the bulk of the crystal. This opens up the possibility for experiments with high-energy particles channeled in a crystalline undulator, a novel compact source of radiation. PMID- 12570494 TI - Temperature-transformed "minimal coupling": magnetofluid unification. AB - The dynamics of a relativistic, hot charged fluid is expressed in terms of a hybrid magnetofluid field which unifies the electromagnetic field with an appropriately defined but analogous flow field. The unification is affected by a well-defined prescription that allows the derivation of the equations of motion of a plasma embedded in an electromagnetic field from the field-free equations. The relationship of this prescription with the minimal coupling prescription of particle dynamics is discussed; the changes brought about by the plasma temperature are highlighted. A few consequences of the unification are worked out. PMID- 12570495 TI - Measurement of internal magnetic field fluctuations in a reversed-field pinch by Faraday rotation. AB - Magnetic field fluctuations (and the associated current perturbation) have been measured in the core of a high-temperature reversed-field pinch using a newly developed fast-polarimetry system. Radial magnetic field fluctuation levels of approximately 1% are measured in standard-reversed-field pinch discharges which increase to approximately 4% during the sawtooth crash (enhanced dynamo). The fluctuation level is reduced fourfold for high-confinement plasmas where the core resonant tearing modes are suppressed. PMID- 12570496 TI - Measurement of the current sheet during magnetic reconnection in a toroidal plasma. AB - The current and magnetic-field fluctuations associated with magnetic-field-line reconnection have been measured in the reversed field pinch plasma configuration. The current sheet resulting from this reconnection has been measured. The current layer is radially broad, comparable to a magnetic-island width, as may be expected from current transport along magnetic-field lines. It is much larger than that predicted by resistive MHD for linear tearing modes and larger than prediction from two-fluid linear theory. PMID- 12570497 TI - Measurements of the shear Alfven wave dispersion for finite perpendicular wave number. AB - Measurements of the dispersion relation for shear Alfven waves as a function of the perpendicular wave number are reported for the regime in which V(A) approximately V(Te). By measuring the parallel phase velocity of the waves, the measurements can be compared directly to theoretical predictions of the dispersion relation for a parameter regime in which particle kinetic effects become important. The comparison shows that the best agreement between theory and experiment is achieved when a fully complex, warm plasma dispersion relation is used. PMID- 12570498 TI - Demonstration of radiation symmetry control for inertial confinement fusion in double Z-pinch hohlraums. AB - Simulations of a double Z-pinch hohlraum, relevant to the high-yield inertial confinement-fusion concept, predict that through geometry design the time integrated P2 Legendre mode drive asymmetry can be systematically controlled from positive to negative coefficient values. Studying capsule elongation, recent experiments on Z confirm such control by varying the secondary hohlraum length. Since the experimental trend and optimum length are correctly modeled, confidence is gained in the simulation tools; the same tools predict capsule drive uniformity sufficient for high-yield fusion ignition. PMID- 12570499 TI - Magnetic shear damping of dissipative drift wave turbulence. AB - The influence of local and global magnetic field line shear on structure formation and transport in dissipative drift-Alfven turbulence is explored. It is found that the generation of zonal flow shear is connected to magnetic shear in ways not accounted previously. The concept of a locally sheared slab flux tube model (including toroidicity) is introduced in order to extend previous analyses to general local variations of magnetic field line shear. It is shown that local shear damping is efficient even when flux surface averaged shear is low. PMID- 12570500 TI - Liquid 3He thermomechanical effect in regular cylinders. AB - We have measured the normal liquid 3He thermomechanical effect with the liquid confined in the regular 210 nm diameter cylinders of Anopore. The pressure difference resulting from the temperature difference was deltaP/deltaT=25 Pa/mK, independent of temperature, from 0.7 to 12 mK. This value was 100x the theoretical value and 3x the result with a packed powder constriction. Two monolayers of 4He did not change the value, but four monolayers reduced it by 2 orders of magnitude; the high value was therefore attributed to boundary scattering from magnetic 3He surface layers. PMID- 12570501 TI - Enhanced Friedel structure and proton pairing in dense solid hydrogen. AB - The mechanism of proton pairing in dense solid hydrogen, and its progression with density, are both studied using effective potentials between protons which include electronic response up to quadratic terms. For high pressures nonlinear effects originating with different pairs are crucial in establishing the net attraction within a given pair, and in this picture Friedel oscillations are considerably enhanced by quadratic response, subsequently playing a very important role in the overall pairing. Calculated vibron frequencies also show substantial agreement with experiment, reflecting at the same time significant changes in the physical character of the pairing itself. PMID- 12570502 TI - Fully coordinated silica nanoclusters: (SiO2)N molecular rings. AB - A new form of finite silica with edge-sharing SiO2 units connected in a ring is proposed. High-level density-functional calculations for (SiO2)(N), N=4-14, show the rings to be energetically more stable than the corresponding (SiO2)(N) linear chains for N>11. The rings display frequency modes in remarkable agreement with infrared bands measured on dehydrated silica surfaces indicating their potential as models of strained extended silica systems. Silica rings, if synthesized, may also be useful precursors for new bulk-silica polymorphs with tubular or porous morphologies. PMID- 12570503 TI - Light scattering study of a twist grain boundary liquid crystal. AB - Dynamic light scattering is used to probe the fluctuation modes of a liquid crystal exhibiting twist grain boundary (TGB) structure. At the chiral nematic to proposed "chiral line liquid" phase transition, anomalous temperature dependence in the fluctuation spectrum and an instability in the helicoidal director structure signify developing TGB order. At lower temperatures, the behavior of the smectic layer-director fluctuations ("soft" mode) indicates that the previously identified commensurate TGB(A) phase may in fact be a TGB(C) phase with an unusually small tilt angle. PMID- 12570504 TI - First-principles calculations of pseudolocal vibrational modes: the case of Cu and Cu pairs in Si. AB - Pseudolocal vibrational modes (pLVMs) are defect-related vibrational modes which are localized despite being below the phonon maximum. Such modes are sometimes seen as phonon replicas in photoluminescence spectra. The pLVMs associated with two copper-related defects are calculated from first-principles density functional theory in periodic supercells. The localization of the pLVMs is quantified using the magnitude of the eigenvectors of the dynamical matrix. PMID- 12570505 TI - Use of a wave reverberation technique to infer the density compression of shocked liquid deuterium to 75 GPa. AB - A novel approach was developed to probe density compression of liquid deuterium (L-D2) along the principal Hugoniot. Relative transit times of shock waves reverberating within the sample are shown to be sensitive to the compression due to the first shock. This technique has proven to be more sensitive than the conventional method of inferring density from the shock and mass velocity, at least in this high-pressure regime. Results in the range of 22-75 GPa indicate an approximately fourfold density compression, and provide data to differentiate between proposed theories for hydrogen and its isotopes. PMID- 12570506 TI - Structure of a collapsed polymer chain with stickers: a single- or multiflower? AB - Using an amphiphilic copolymer with evenly spaced hydrophobic styrene segments (stickers), poly(N-isopropylacrylamide-s-styrene), we recently confirmed a long standing prediction that such a copolymer chain in a solvent selectively poor for the stickers could self-fold from a random coil to a single-flowerlike core-shell nanostructure. Moreover, we found that the self-folding involves the movement of the sticks to the center and the transition from the random coil to the collapsed globule passes through a proposed ordered coil state. PMID- 12570507 TI - Endohedral impurities in carbon nanotubes. AB - A generalization of the Anderson model that includes pseudo-Jahn-Teller impurity coupling is proposed to describe distortions of an endohedral impurity in a carbon nanotube. Within mean-field theory, spontaneous axial symmetry breaking is found when the vibronic coupling strength g exceeds a critical value. The effective potential is found to have O(2) symmetry, in agreement with numerical calculations. For metallic zigzag nanotubes endohedrally doped with transition metals in the dilute limit, the low-energy properties of the system may display two-channel Kondo behavior; however, strong vibronic coupling is seen to exponentially suppress the Kondo energy scale. PMID- 12570508 TI - Fast calculation of the density of states of a fluid by Monte Carlo simulations. AB - Two related methods are proposed to calculate the density of states of a fluid from Monte Carlo simulations. In contrast to previous approaches, which require that histograms be accumulated in a stochastic manner, the methods proposed here rely on evaluation of the instantaneous temperature. In the first method, the temperature is calculated from the gradient of the forces. In the second, it is estimated from the kinetic contribution to the total energy. The validity and usefulness of the new approaches are demonstrated by presenting results from simulations of a Lennard-Jones fluid. It is shown that the new methods are considerably faster than previously available techniques. PMID- 12570509 TI - Effect of impurities in the large Au-Au distances in gold nanowires. AB - Experimentally obtained atomically thin gold nanowires have presented exceedingly large Au-Au interatomic distances before they break. Since no theoretical calculations of pure gold nanowires have been able to produce such large distances, we have investigated, through ab initio calculations, how impurities could affect them. We have studied the effect of H, B, C, N, O, and S impurities on the nanowire electronic and structural properties, in particular how they affect the maximum Au-Au bond length. We find that the most likely candidates to explain the distances in the range of 3.6 A and 4.8 A are H and S impurity atoms, respectively. PMID- 12570510 TI - Ion beam triangulation of ultrathin Mn and CoMn films grown on Cu(001). AB - Total target currents for grazing scattering of keV protons from a crystal target are used to investigate the structure of surfaces and ultrathin films. This current shows pronounced maxima whenever the azimuthal incidence angle coincides with close-packed rows of atoms in the surface and subsurface layers. The real space method is applied to study monolayer and bilayer films of Mn and of CoMn epitaxially grown on a Cu(001) surface. PMID- 12570511 TI - Measurements of elastic constants in thin films of colossal magnetoresistance material. AB - Measurements of elastic constants of strained 200 and 400 nm thin films, as well as unstrained samples, of the colossal magnetoresistance (CMR) material La0.67Ca0.33MnO3 are presented. Since the peak resistance temperature of a strained CMR film decreases as the film thickness decreases, it is of interest to see if features in the elastic constants, reflecting structural or magnetic changes, follow the peak resistance temperature. It is observed that features in the elastic constants appear not only at the peak resistance temperatures of the CMR samples, but also at a temperature about 17 K higher. A new technique, thin film resonant ultrasound spectroscopy, was used to make the measurements. PMID- 12570512 TI - First-principles calculation of vibrational Raman spectra in large systems: signature of small rings in crystalline SiO2. AB - We present an approach for the efficient calculation of vibrational Raman intensities in periodic systems within density functional theory. The Raman intensities are computed from the second order derivative of the electronic density matrix with respect to a uniform electric field. In contrast to previous approaches, the computational effort required by our method for the evaluation of the intensities is negligible compared to that required for the calculation of vibrational frequencies. As a first application, we study the signature of 3- and 4-membered rings in the Raman spectra of several polymorphs of SiO2, including a zeolite (H-ZSM-18) having 102 atoms per unit cell. PMID- 12570513 TI - Triplet-to-singlet exciton formation in poly(p-phenylene-vinylene) light-emitting diodes. AB - The triplet to singlet exciton formation ratio in a MEH-PPV light-emitting diode is measured by comparing the triplet-induced absorptions with optical and electric excitations at the same singlet exciton density. The ratio is a strong universal decreasing function of the averaged electric field. Using 4 ns for singlet to triplet intersystem crossing time, the ratio is significantly larger than the spin-independent value 3 at intermediate field but is reduced to about 2 for higher field. PMID- 12570514 TI - Second-harmonic generation in GaAs: experiment versus theoretical predictions of chi(2)xyz. AB - For GaAs we have determined |chi((2))(xyz)(-2omega;omega,omega)| in second harmonic generation experiments using two-photon energies between 2 and 5 eV. In addition to the E1, E1+delta(1), E'0, and E2 critical-point bulk transitions of GaAs, a surprisingly strong surface transition at 3.35 eV was observed for natively oxidized GaAs(001) samples. A detailed comparison with theoretical predictions reveals that calculations that include many-particle effects at the level of the "scissors" approximation can describe the overall frequency dependence of the second-harmonic susceptibility reasonably well. PMID- 12570515 TI - Artificial dielectric superlattices with broken inversion symmetry. AB - A variety of three-constituent superlattice patterns were made in atomic layer-by layer films, with patterns breaking inversion symmetry giving effective permanent bias fields ranging up to about 200 kV/cm. Dielectric constants at room temperature were nearly 10(3), with loss tangents under 0.01. Most of the response came from discrete dipoles comprising multiple unit cells, but without any ferroelectric phase transition. PMID- 12570516 TI - Evidence of Landau levels and interactions in low-lying excitations of composite fermions at 1/3or=nu>or=1/3 of the fractional quantum Hall regime are observed by resonant inelastic light scattering. Spectra of spin reversed excitations suggest a structure of lowest spin-split Landau levels of composite fermions that is similar to that of electrons. Spin-flip energies determined from spectra reveal significant composite fermion interactions. The filling factor dependence of mode energies displays an abrupt change in the middle of the range when there is partial population of a composite fermion level. PMID- 12570517 TI - Dynamical matching of Josephson vortex lattice with sample edge in layered high Tc superconductors: origin of the periodic oscillation of flux flow resistance. AB - We numerically investigate Josephson vortex flow states in layered high-T(c) superconductors motivated by a recent experimental observation for accurate periodic magnetic field dependences of the Josephson vortex flow resistance over a wide range of magnetic field (0.5-4.0 T). We confirm in our mesoscale simulations that dynamical matching of Josephson vortex lattice with sample edge is responsible for the periodic dependence. The present simulations reveal that the Josephson vortex lattice flow speed is particularly suppressed when the moment of vortex entry matches that of vortex escape. Thus, the possible matching situations are taken into account and the observed periodicity is successfully explained. PMID- 12570518 TI - Phase diagram of La2-xSrxCuO4 probed in the infared: imprints of charge stripe excitations. AB - While there is increasing evidence for antiferromagnetic (AF) ordering in the Cu O planes of high-T(c) superconductors, either static or fluctuating, there is no direct evidence so far for the charge stripes that should separate the AF domains. By investigating the optical response of La2-xSrxCuO4 for 0X(s)l+l-. AB - We report the first measurement of the branching fraction for the inclusive decay B-->X(s)l(+)l(-), where l is either an electron or a muon, and X(s) is a hadronic recoil system that contains an s quark. We analyzed a data sample of 65.4 x 10(6) B meson pairs collected with the Belle detector at the KEKB e(+)e(-) asymmetric energy collider. We find B(B-->X(s)l(+)l(-))=[6.1 +/-1.4(stat) +1.4-1.1(syst)] x 10(-6) for dilepton masses greater than 0.2 GeV/c(2). PMID- 12570536 TI - First results from KamLAND: evidence for reactor antineutrino disappearance. AB - KamLAND has measured the flux of nu;(e)'s from distant nuclear reactors. We find fewer nu;(e) events than expected from standard assumptions about nu;(e) propagation at the 99.95% C.L. In a 162 ton.yr exposure the ratio of the observed inverse beta-decay events to the expected number without nu;(e) disappearance is 0.611+/-0.085(stat)+/-0.041(syst) for nu;(e) energies >3.4 MeV. In the context of two-flavor neutrino oscillations with CPT invariance, all solutions to the solar neutrino problem except for the "large mixing angle" region are excluded. PMID- 12570540 TI - Universal pion freeze-out in heavy-ion collisions. AB - Based on an evaluation of data on pion interferometry and on particle yields at midrapidity, we propose a universal condition for thermal freeze-out of pions in heavy-ion collisions. We show that freeze-out occurs when the mean free path of pions lambda(f) reaches a value of about 1 fm, which is much smaller than the spatial extent of the system at freeze-out. This critical mean free path is independent of the centrality of the collision and beam energy from the Alternating Gradient Synchrotron to the Relativistic Heavy Ion Collider. PMID- 12570542 TI - Echo spectroscopy and quantum stability of trapped atoms. AB - We investigate the dephasing of ultra cold 85Rb atoms trapped in an optical dipole trap and prepared in a coherent superposition of their two hyperfine ground states by interaction with a microwave pulse. We demonstrate that the dephasing, measured as the Ramsey fringe contrast, can be reversed by stimulating a coherence echo with a pi pulse between the two pi / 2 pulses, in analogy to the photon echo. We also demonstrate that "echo spectroscopy" can be used to study the quantum dynamics in the trap even when more than 10(6) states are thermally populated and to study the crossover from quantum to classical dynamics. PMID- 12570541 TI - Precision measurement of the 7Be(p,gamma)8B cross section with an implanted 7Be target. AB - The 7Be(p,gamma)8B reaction plays a central role in the evaluation of solar neutrino fluxes. We report on a new precision measurement of the cross section of this reaction, following our previous experiment with an implanted 7Be target, a raster-scanned beam, and the elimination of the backscattering loss. The new measurement incorporates a more abundant 7Be target and a number of improvements in design and procedure. The point at E(lab)=991 keV was measured several times under varying experimental conditions, yielding a value of S17(E(c.m.)=850 keV)=24.0+/-0.5 eV b. Measurements were carried out at lower energies as well. Because of the precise knowledge of the implanted 7Be density profile, it was possible to reconstitute both the off- and on-resonance parts of the cross section and to obtain from the entire set of measurements an extrapolated value of S17(0)=21.2+/-0.7 eV b. PMID- 12570543 TI - Circular polarization of ion fluorescence completing the analysis of resonant Xe* 4d(-1)(5/2)6p Auger decay. AB - The relative contributions of the partial electron waves emitted in the Auger decay of the Xe* 4d(-1)(5/2)6p(J(*)=1) resonance have been determined by fluorescence polarimetry after excitation with circularly polarized synchrotron radiation. The analysis of circularly polarized fluorescence of the photoion leads to an independent determination of the orientation parameters for all states of the Xe II 5p(4)6p multiplet. The present study provides, in combination with data on the angular distribution and spin polarization of the Auger electrons, complete quantum mechanical information on the resonant Auger decay, i.e., branching ratios and relative phases of the Auger decay amplitudes. PMID- 12570544 TI - High-density trapping of cold ytterbium atoms by an optical dipole force. AB - We have succeeded in trapping a high density of rare-earth atom of ytterbium (Yb) in a crossed far-off resonance trap. The peak density reaches more than 10(14) cm(-3). With a new method of a delayed crossed far-off resonance trap, we have elucidated that the atoms became concentrated into the cross region by atom-atom collisions. We trap fermionic Yb atoms in the same way as bosonic ones. PMID- 12570545 TI - Time-resolved explosion of intense-laser-heated clusters. AB - We investigate the femtosecond explosive dynamics of intense laser-heated argon clusters by measuring the cluster complex transient polarizability. The time evolution of the polarizability is characteristic of competition in the optical response between supercritical and subcritical density regions of the expanding cluster. The results are consistent with time-resolved Rayleigh scattering measurements, and bear out the predictions of a recent laser-cluster interaction model [H. M. Milchberg, S. J. McNaught, and E. Parra, Phys. Rev. E 64, 056402 (2001)]]. PMID- 12570539 TI - Photoproduction of the omega meson on the proton at large momentum transfer. AB - The differential cross section, dsigma/dt, for omega meson exclusive photoproduction on the proton above the resonance region (2.6P in unsteady, transitional flows. We find in the nonlinear flow regime the decay as approximately t(-alpha) where alpha is related to a space-time scaling parameter psi as alpha proportional, variant psi(m), for the considered parameter range m=0.067. In the linear flow regime, we find that the maximum is alpha approximately 2/3 for psi approximately 1. The proposed scaling law should be useful for linking dynamical subgrid processes with reaction kinetics in a variety of transitional flow systems. PMID- 12570586 TI - Bragg rods and multiple X-ray scattering in random-stacking colloidal crystals. AB - Synchrotron small-angle x-ray diffraction images of random-stacking-induced Bragg scattering rods are obtained in a wide range of wave vectors from a single colloidal crystal. The results reveal a strong multiple scattering effect, which leads to new features in the diffraction pattern-secondary Bragg rods. We argue that dynamic x-ray diffraction is rather common for high-quality colloidal photonic crystals and should be taken into account. PMID- 12570587 TI - Absence of epidemic threshold in scale-free networks with degree correlations. AB - Random scale-free networks have the peculiar property of being prone to the spreading of infections. Here we provide for the susceptible-infected-susceptible model an exact result showing that a scale-free degree distribution with diverging second moment is a sufficient condition to have null epidemic threshold in unstructured networks with either assortative or disassortative mixing. Degree correlations result therefore irrelevant for the epidemic spreading picture in these scale-free networks. The present result is related to the divergence of the average nearest neighbor's degree, enforced by the degree detailed balance condition. PMID- 12570588 TI - Comment on "Resonant spectra and the time evolution of the survival and nonescape probabilities". PMID- 12570590 TI - Comment on "Effects of disorder on ferromagnetism in diluted magnetic semiconductors". PMID- 12570592 TI - Comment on "Wave refraction in negative-index media: always positive and very inhomogeneous". PMID- 12570594 TI - Stability and phase coherence of trapped 1D Bose gases. AB - We discuss stability and phase coherence of 1D trapped Bose gases and find that inelastic decay processes, such as three-body recombination, are suppressed in the strongly interacting (Tonks-Girardeau) and intermediate regimes. This is promising for achieving these regimes with a large number of particles. "Fermionization" of the system reduces the phase coherence length, and at T=0 the gas is fully phase coherent only deeply in the weakly interacting (Gross Pitaevskii) regime. PMID- 12570595 TI - Instantaneous measurement of nonlocal variables. AB - It is shown, under the assumption of the possibility to perform an arbitrary local operation, that all nonlocal variables related to two or more separate sites can be measured instantaneously, except for a finite time required for bringing to one location the classical records from these sites which yield the result of the measurement. It is a verification measurement: it yields reliably the eigenvalues of the nonlocal variables, but it does not prepare the eigenstates of the system. PMID- 12570596 TI - Decoherence-induced continuous pointer states. AB - We investigate the reduced dynamics in the Markovian approximation of an infinite quantum spin system linearly coupled to a phonon field at positive temperature. The achieved diagonalization leads to a selection of the continuous family of pointer states corresponding to a configuration space of the one-dimensional Ising model. Such a family provides a mathematical description of an apparatus with continuous readings. PMID- 12570597 TI - Are there phase transitions in information space? AB - The interplay between two basic quantities--quantum communication and information -is investigated. Quantum communication is an important resource for quantum states shared by two parties and is directly related to entanglement. Recently, the amount of local information that can be drawn from a state has been shown to be closely related to the nonlocal properties of the state. Here we consider both formation and extraction processes, and analyze informational resources as a function of quantum communication. The resulting diagrams in information space allow us to observe phaselike transitions when correlations become classical. PMID- 12570598 TI - Levy flights in the Landau-Teller model of molecular collisions. AB - We consider the Landau-Teller model, which is a prototype for the exchanges of energy, in molecular collisions, between internal degrees of freedom and those of the center of mass. We show that the statistics of the energy exchanges computed through the dynamics over a finite time is of the Levy type for high enough frequencies of the internal motions, while it reduces to the familiar Gaussian one in the limit of low frequencies. The relevance for the definition of the times of relaxation to equilibrium is also pointed out. PMID- 12570599 TI - Radiative cycle with stimulated emission from atoms and ions in an astrophysical plasma. AB - We propose that a radiative cycle operates in atoms and ions located in a rarefied gas in the vicinity of a hot star. Besides spontaneous transitions the cycle includes a stimulated transition in one very weak intermediate channel. This radiative "bottleneck" creates a population inversion, which for an appropriate column density results in amplification and stimulated radiation in the weak transition. The stimulated emission opens a fast decay channel leading to a fast radiative cycle in the atom (or ion). We apply this model by explaining two unusually bright Fe II lines at 250.7 and 250.9 nm in the UV spectrum of gas blobs close to eta Carinae, one of the most massive and luminous stars in the Galaxy. PMID- 12570600 TI - Two-loop renormalization group equations in the standard model. AB - Two-loop renormalization group equations in the standard model are recalculated. A new coefficient is found in the beta function of the quartic coupling and a class of gauge invariants is found to be absent in the beta functions of hadronic Yukawa couplings. The two-loop beta function of the Higgs mass parameter is presented in complete form. PMID- 12570601 TI - Consistent resolution of possible anomalies in B0-->phi K(S) and B+ --> eta'K+ Decays. AB - In the framework of R-parity violating (R(p)) supersymmetry, we try to find a consistent explanation for both recently measured CP asymmetry in B0 --> phi K(S) decay and the large branching ratio of B+/- --> eta'K+/- decay, which are inconsistent with the standard model (SM) prediction. We also investigate other charmless hadronic B -- >PP and B --> VP decay modes whose experimental data favor the SM: for instance, recently measured CP asymmetries in B0 --> eta(')K(S) and B0 -- >J/Psi K(S). We find that all the observed data can be accommodated for certain values of R(p) couplings. PMID- 12570602 TI - Double deeply virtual Compton scattering off the nucleon. AB - We study the double deeply virtual Compton scattering (DDVCS) process off the nucleon, through the scattering of a space-like virtual photon with large virtuality resulting in the production of a time-like virtual photon, decaying into an e(+)e(-) pair. This process is expressed in the Bjorken regime in terms of generalized parton distributions (GPDs), and it is shown that by varying the invariant mass of the lepton pair, one can directly extract the GPDs from the observables. We give predictions for the DDVCS cross section and beam helicity asymmetry and discuss its experimental feasibility. PMID- 12570603 TI - Novel search for heavy nu mixing from the beta+ decay of 38mK confined in an atom trap. AB - A new technique, full neutrino momentum reconstruction, is used to set limits on the admixture of heavy neutrinos into the electron neutrino. We measure coincidences between nuclear recoils and positrons from the beta decay of trapped radioactive atoms and deduce the neutrino momentum. A search for peaks in the reconstructed recoil time-of-flight spectrum as a function of positron energy is performed. The admixture upper limits range from 4 x 10(-3) to 2 x 10(-2) and are the best direct limits for neutrinos (as opposed to antineutrinos) for the mass region of 0.7 to 3.5 MeV. PMID- 12570604 TI - Fine structure in proton emission from 145Tm discovered with digital signal processing. AB - Fine structure in proton emission from the 3.1(3) mus activity of 145Tm was discovered by using a novel technique of digital processing of overlapping recoil implantation and decay signals. Proton transitions to the ground state of 144Er and to its first excited 2(+) state at 0.33(1) MeV with a branching ratio I(p)(2(+))=9.6+/-1.5% were observed. The structure of the 145Tm wave function and the emission process were analyzed by using particle-core vibration coupling models. PMID- 12570605 TI - Conical intersections and bound molecular states embedded in the continuum. AB - Nuclear dynamics on coupled potential surfaces can lead to bound states embedded in the continuum. For one type of conical intersection situation, an explicit proof is presented that such states exist. Non-Born-Oppenheimer effects are responsible for the binding of these states. Once the Born-Oppenheimer approximation is introduced, these states at best become resonances which decay via potential tunneling. The tunneling is completely suppressed by the coupling between the electronic states. A numerical example is given. PMID- 12570606 TI - Photodetachment of He- in the vicinity of the two-electron escape threshold. AB - Recording the yield of He(1snl(3)L) Rydberg states for n=11-14, we measure the photodetachment cross sections of metastable He-(1s2s2p(4)P(o)) ions in the vicinity of the two-electron escape threshold. We observe a large number of double Rydberg He- quartet state resonances and report energies and widths of intrashell states in the n=13-15 manifolds. Sharp thresholds are measured at He((3)P(o)) and He((3)D(e)) Rydberg states with preference for population of the former, whereas the He((3)S(e)) states are not populated, in agreement with qualitative theoretical arguments. PMID- 12570607 TI - Strong field atomic ionization: origin of high-energy structures in photoelectron spectra. AB - Two distinct interpretations have been proposed to account for conspicuous enhancements of the ionization peaks in the high energy part of above-threshold ionization spectra. One of them ascribes the enhancement to a multiphoton resonance involving an excited state, while other analysis performed for zero range model potential link it to "channel closings, " i.e., to the change in the number of photons needed to ionize the atom when the laser intensity increases. We report the results of model calculations that confirm the existence of a resonant process in atoms and shed light on why short-range potential models can mimic the experimental observations. PMID- 12570608 TI - Single-molecule study of the electronic couplings in a circular array of molecules: light-harvesting-2 complex from Rhodospirillum molischianum. AB - Applying single-molecule spectroscopic techniques allowed us to determine the mutual angles between the transition-dipole moments associated with optical transitions of the eight bacteriochlorophyll a molecules which form the so-called B800 ring of the light-harvesting-2 complex from Rhodospirillum molischianum. The orientation of the transition-dipole moment is a sensitive probe for the strength of the local intermolecular interactions because of the well-defined arrangement of the individual molecules within the B800 ring. Our data reveal that the strength of the electronic coupling between individual molecules in the ring is subjected to spatial as well as temporal variations. PMID- 12570609 TI - Thermally enhanced neutralization in hyperthermal energy ion scattering. AB - Neutralization probabilities are presented for hyperthermal energy Na+ ions scattered from a Cu(001) crystal as a function of surface temperature and scattered velocity. A large enhancement in neutralization is observed as the temperature is increased. Velocity-dependent charge transfer regimes are probed by varying the incident energy, with the most prominent surface temperature effects occurring at the lowest energies. The data agree well with results obtained from a model based on the Newns-Anderson Hamiltonian, where the effects of both temperature and velocity are incorporated. PMID- 12570610 TI - Eigenvalues of the Zakharov-Shabat scattering problem for real symmetric pulses. AB - The classical problem of determining the solitons generated from symmetric real initial conditions in the nonlinear Schrodinger equation is revisited. The corresponding Zakharov-Shabat scattering problem is solved for real and symmetric double-humped rectangular initial pulse forms. It is found that such real symmetric pulses may generate eigenvalues with nonzero real parts corresponding to separating soliton pulse pairs. Moreover, it is found that the classical formula relating the number of eigenvalues to the area of the pulse is not always correct. PMID- 12570611 TI - Nonlinear theory of polarization-mode dispersion for fiber solitons. AB - We consider the evolution of optical solitons inside a nonlinear dispersive fiber with random birefringence, causing polarization-mode dispersion. We convert the pair of coupled nonlinear Schrodinger equations satisfied by the orthogonally polarized components into a Fokker-Planck equation using the collective-variable approach. We solve this equation and derive expressions for the probability density functions associated with the differential group delay and the pulse width in the limit of large propagation distances. PMID- 12570612 TI - Near-field second-harmonic generation induced by local field enhancement. AB - The field near a sharp metal tip can be strongly enhanced if irradiated with an optical field polarized along the tip axis. We demonstrate that the enhanced field gives rise to local second-harmonic (SH) generation at the tip surface thereby creating a highly confined photon source. A theoretical model for the excitation and emission of SH radiation at the tip is developed and it is found that this source can be represented by a single on-axis oscillating dipole. The model is experimentally verified by imaging the spatial field distribution of strongly focused laser modes. PMID- 12570613 TI - X-ray interferometry with microelectronvolt resolution. AB - We demonstrate an interferometer for hard x rays with two back-reflecting sapphire crystal mirrors--a prototype x-ray Fabry-Perot interferometer. A finesse of 15 and 0.76 mu eV broad Fabry-Perot transmission resonances are measured by the time response of the interferometer. Interference patterns are observed directly in spectral dependences of reflectivity. PMID- 12570614 TI - Self-organized and driven phase synchronization in coupled maps. AB - We study the phase synchronization and cluster formation in coupled maps on different networks. We identify two different mechanisms of cluster formation: (a) self-organized phase synchronization which leads to clusters with dominant intracluster couplings and (b) driven phase synchronization which leads to clusters with dominant intercluster couplings. In the novel driven synchronization the nodes of one cluster are driven by those of the others. We also discuss the dynamical origin of these two mechanisms for small networks with two and three nodes. PMID- 12570615 TI - Phase shift experiments identifying Kramers doublets in a chaotic superconducting microwave billiard of threefold symmetry. AB - The spectral properties of a two-dimensional microwave billiard showing threefold symmetry have been studied with a new experimental technique. This method is based on the behavior of the eigenmodes under variation of a phase shift between two input channels, which strongly depends on the symmetries of the eigenfunctions. Thereby a complete set of 108 Kramers doublets has been identified by a simple and purely experimental method. This set clearly shows Gaussian unitary ensemble statistics, although the system is time-reversal invariant. PMID- 12570616 TI - Parameter scaling in the decoherent quantum-classical transition for chaotic systems. AB - The quantum to classical transition for a system depends on many parameters, including a scale length for its action, variant Planck's over 2 pi, a measure of its coupling to the environment, D, and, for chaotic systems, the classical Lyapunov exponent, lambda. We propose measuring the proximity of quantum and classical evolutions as a multivariate function of (Planck's over 2 pi,lambda,D) and searching for transformations that collapse this hypersurface into a function of a composite parameter zeta= Planck's over 2 pi alpha)lambda beta D gamma. We report results for the quantum Cat Map and Duffing oscillator, showing accurate scaling behavior over a wide parameter range, indicating that this may be used to construct universality classes for this transition. PMID- 12570617 TI - Taking advantage of multiple scattering to communicate with time-reversal antennas. AB - We present an experimental demonstration showing that, contrary to first intuition, the more scattering a mesoscopic medium is, the more information can be conveyed through it. We used a multiple input-multiple output configuration: a multichannel ultrasonic time-reversal antenna is used to transmit random series of bits simultaneously to different receivers which were only a few wavelengths apart. Whereas the transmission is free of error when multiple scattering occurs in the propagation medium, the error rate is huge in a homogeneous medium. PMID- 12570618 TI - Reversing the Brazil-nut effect: competition between percolation and condensation. AB - We report on experiments on vertically shaken binary granular mixtures, which separate into their components due to the external excitation. This well-known phenomenon, where large particles rise to the top of the mixture, is called the Brazil-nut effect. Recent theoretical findings predict also a reverse Brazil-nut effect, where large particles sink to the bottom of the container. We choose spherical beads of various diameters and materials in order to observe the transition from Brazil-nut effect to its reverse form. The direction of demixing depends sensitively on the external excitation, so that it is possible to switch between both effects for a given mass density ratio. PMID- 12570619 TI - Transition to turbulence in particulate pipe flow. AB - We investigate experimentally the influence of suspended particles on the transition to turbulence. The particles are monodisperse and neutrally buoyant with the liquid. The role of the particles on the transition depends upon both the pipe to particle diameter ratios and the concentration. For large pipe-to particle diameter ratios the transition is delayed while it is lowered for small ratios. A scaling is proposed to collapse the departure from the critical Reynolds number for pure fluid as a function of concentration into a single master curve. PMID- 12570620 TI - Quadrupole-induced resonant-particle transport in a pure electron plasma. AB - Small transverse magnetic quadrupole fields sharply degrade the confinement of non-neutral plasmas held in Malmberg-Penning traps. For example, a quadrupole magnetic field of only 0.02 G/cm doubles the diffusion rate in a trap with a 100 G axial magnetic field. Larger quadrupole fields noticeably change the shape of the plasma. The transport is greatest at an orbital resonance. These results cast doubt on plans to use magnetic quadrupole neutral atom traps to confine antihydrogen atoms created in double-well positron/antiproton Malmberg-Penning traps. PMID- 12570621 TI - Transport reduction by rotation shear in tokamak-edge turbulence. AB - Effects of externally imposed and self-generated poloidal flows on turbulent transport in the edge region of a tokamak are investigated using 3D nonlinear global simulations of resistive pressure-gradient-driven turbulence. Transport reduction is found to be due to synergetic changes in the fluctuation amplitude and in the dephasing of the fluctuations. A scaling of the fluctuation level and turbulent diffusivity with E x B flow shear strength is deduced from these simulations. These scalings agree with recent experimental observations on edge biasing as well as with analytical models. PMID- 12570622 TI - Stability of small carbon-nitride heterofullerenes. AB - Carbon nitride nanostructures have been produced by the arc-discharge technique and analyzed by mass spectrometry. A series of structured peaks in the region of masses from 480 up to 600 suggests the existence of heterofullerenes C(n-x)Nx(40 < or = n < or = 50). The structure and stability of these small fullerenes were theoretically investigated by quantum chemical calculations. The obtained heats of formation indicate that C(n) molecules stabilize upon nitrogen substitution. Two C(n-x)Nx cages are quite stable, with heats of formation per atom approaching that of C60. These molecules could be the seeds of onion-like structures seen in CN materials [Phys. Rev. Lett. 87, 225503 (2001)]]. PMID- 12570623 TI - Hollow-core dendrimers revisited. AB - In revisiting the original argument of de Gennes and Hervet [J. Phys. (Paris) 44, L351 (1983)]] leading to "hollow-core" dendrimers, we show that a self-consistent application of their model leads to the "filled-core" model first elucidated by Lescanec and Muthukumar [Macromolecules 23, 2280 (1990)]]. The monomer density falls off parabolically from the center of the dendrimer, and the tips are distributed throughout the molecule. PMID- 12570624 TI - Partial crystallization of an amorphous alloy by electronic energy deposition. AB - The first experimental evidence is reported of crystallization induced in an amorphous alloy by a high density of electronic excitation deposited along the path of swift heavy ions. The formation of nanocrystalline iron boride phases was observed in an amorphous Fe73.5Cu1Nb3Si13.5B9 alloy irradiated at low temperature with 5 GeV Pb ions up to fluences of 1 x 10(11) ions cm-2. No evidence for the formation of the Fe(Si) phase was found. This phenomenon was interpreted in terms of the relaxation of the high level of energy deposited in electronic excitations along the path of Pb ions in the target, which induces extensive stress and strain that could destabilize the amorphous structure. PMID- 12570625 TI - Surface ordering transitions at a liquid crystal-solid interface above the isotropic smectic-A transition. AB - The degree of orientational order induced by confining cylindrical surfaces is monitored via deuteron nuclear magnetic resonance linesplitting and linewidth above the smectic-A to isotropic phase transition. The orientational order strongly depends on the length of the surfactant coupling molecule, on the surface coverage, and on the liquid crystal. Continuous and stepwise growth of orientational order and surface-induced orientational order transitions found in the isotropic phase are explained in terms of a simplified model of surface induced layering and molecular self-diffusion. PMID- 12570626 TI - Self-diffusion of tris-naphthylbenzene near the glass transition temperature. AB - We present a direct measurement of self-diffusion of a single-component glass forming liquid at the glass transition temperature. Forward recoil spectrometry is used to measure the concentration profiles of deuterio and protio 1,3-bis-(1 naphthyl)-5-(2-naphthyl)benzene (TNB) following annealing-induced diffusion in a vapor-deposited bilayer. These experiments extend the range of measured diffusion coefficients in TNB by 6 orders of magnitude. The results indicate a decoupling of translational diffusion coefficients from viscosity or rotation. At T(g), D(T) is 400 times larger than expected from the Stokes-Einstein equation. PMID- 12570627 TI - Two-dimensional coarsening kinetics of reconstruction domains: GaAs(001)-beta(2 x 4). AB - We study the nonconserved coarsening kinetics of a reconstructed semiconductor surface. The domain size evolution is obtained in situ by time-resolved surface x ray diffraction. The system exhibits four equivalent domain types with two nonequivalent types of domain boundaries. Small domains are prepared by molecular beam epitaxy deposition of one GaAs layer. We find the correlation lengths of the domain size distribution to depend on time as l is proportional to t(0.42+/-0.05) in the half-order reflections and l is proportional to t(0.22+/-0.05) in the quarter-order reflections. The fraction of the higher energy domain boundaries increases as lnt. PMID- 12570628 TI - Novel stabilization mechanism on polar surfaces: ZnO(0001)-Zn. AB - The (1x1) terminated (0001)-Zn surface of wurtzite ZnO was investigated with scanning tunneling microscopy. The surface is characterized by the presence of nanosized islands with a size-dependent shape and triangular holes with single height, O-terminated step edges. It is proposed that the resulting overall decrease of the surface Zn concentration stabilizes this polar surface. Ab initio calculations of test geometries predict triangularly shaped reconstructions over a wide range of oxygen and hydrogen chemical potentials. The formation of these reconstructions appears to be electrostatically driven. PMID- 12570629 TI - Oxidation at the Si/SiO2 interface: influence of the spin degree of freedom. AB - We show, using first-principles spin-polarized total-energy calculations, that depending on the spin configuration of the system, the reaction of an O2 molecule with a Si-Si bond in a suboxidized region might result either in a peroxy linkage defect (for a singlet spin state) or in a perfect Si-O-Si bond plus an interstitial O atom (for a triplet spin state). Even though the singlet has a lower energy than the triplet configuration, we find a rather small probability for triplet to singlet conversion. Therefore, as the O2 in an SiO2 interstitial site has a triplet configuration, this reaction spin dependence may have a strong influence on the high quality of the Si/SiO2 interface. PMID- 12570630 TI - Rapid roughening in thin film growth of an organic semiconductor (diindenoperylene). AB - The scaling exponents alpha, beta, and 1/z in thin films of the organic molecule diindenoperylene deposited on SiO2 under UHV conditions are determined. Atomic force microscopy, x-ray reflectivity, and diffuse x-ray scattering were employed. The surface width displays power law scaling over more than 2 orders of magnitude in film thickness. We obtained alpha = 0.684+/-0.06, beta = 0.748+/-0.05, and 1/zeta = 0.92+/-0.20. The derived exponents point to an unusually rapid growth of vertical roughness and lateral correlations. We suggest that they could be related to lateral inhomogeneities arising from the formation of grain boundaries between tilt domains in the early stages of growth. PMID- 12570631 TI - Hydrodynamic origin of diffusion in nanopores. AB - We study the transport of a subcritical Lennard-Jones fluid in a cylindrical nanopore, using a combination of equilibrium and nonequilibrium as well as dual control volume grand canonical molecular dynamics methods. We show that all three techniques yield the same value of the transport coefficient for diffusely reflecting pore walls, even in the presence of viscous transport. We also demonstrate that the classical Knudsen mechanism is not manifested, and that a combination of viscous flow and momentum exchange at the pore wall governs the transport over a wide range of densities. PMID- 12570632 TI - Bridging simulations and experiments in microstructure evolution. AB - We demonstrate the importance of anisotropic interface properties in microstructure evolution by comparing computed evolved microstructures to final experimental microstructures of 5170 grains in 19 thin aluminum foil samples. This is the first time that a direct experimental validation of simulation has been performed at the level of individual grains. We observe that simulated microstructures using curvature-driven grain boundary motion and anisotropic interface properties agree well with experimentally evolved microstructures, whereas agreement is poor when isotropic properties are used. PMID- 12570633 TI - Measuring local optical properties: near-field polarimetry of photonic block copolymer morphology. AB - Ultrahigh molecular weight polystyrene-b-polyisoprene block copolymers (BCs), noted for their photonic behavior, were imaged using transmission near-field scanning optical microscopy (NSOM) and NSOM polarimetry. Our improved scheme for polarization modulation (PM) polarimetry, which accounts for optical anisotropies of the NSOM aperture probe, enables mapping of the local diattenuation and birefringence (with separately aligned diattenuating and fast axes) in these specimens with subdiffraction limited resolution. PM-NSOM micrographs illuminate the mesoscopic optical nature of these BC specimens by resolving individual microphase domains and defect structures. PMID- 12570634 TI - Narrow-gap Luttinger liquid in carbon nanotubes. AB - Electron interactions reinforce minigaps induced in metallic nanotubes by an external field and turn the gap field dependence into a universal power law. An exactly solvable Gross-Neveau model with an SU(4) symmetry is derived for neutral excitations near half filling. Charge excitations, described by a sine-Gordon perturbation of Luttinger liquid theory, are composite solitons formed by the charged and neutral fields with two separate length scales. Charge compressibility at finite density, evaluated in terms of intersoliton interaction, exhibits a crossover from overlapping to nonoverlapping soliton state. Implications for the Coulomb blockade measurements are discussed. PMID- 12570635 TI - Glassy behavior of electrons near metal-insulator transitions. AB - The emergence of glassy behavior of electrons is investigated for systems close to disorder-driven and interaction-driven metal-insulator transitions. Our results indicate that Anderson localization effects strongly stabilize such glassy behavior, while Mott localization tends to suppress it. We predict the emergence of an intermediate metallic glassy phase separating the insulator from the normal metal. This effect is expected to be most pronounced for sufficiently disordered systems, in agreement with recent experimental observations. PMID- 12570636 TI - Indication of charge-density-wave formation in Bi(111). AB - Photoemission spectroscopy of Bi(111) reveals a small hexagonal two-dimensional Fermi surface (FS) associated with an electron band centered in the surface Brillouin zone. Along the hexagon the Fermi momentum k(F) ranges from 0.053 to 0.061 A(-1). Temperature dependent valence band spectra show an anisotropic energy gap Delta near the Fermi level. We find a transition temperature of about 75 K. At 11 K, the gap is Delta=4 meV at the corner and Delta=7.5 meV at the side of the hexagon. Arguments based on susceptibility chi(--> q) calculations of a hexagonal FS are used to discuss an incommensurate charge-density-wave (CDW) formation associated with a q(CDW)=0.106 A(-1). PMID- 12570637 TI - Origin of the residual acceptor ground-state splitting in silicon. AB - The residual ground-state splitting of acceptors in high-quality silicon has been studied intensely by different experimental techniques for several decades. Recently, photoluminescence studies of isotopically pure silicon revealed the ground-state splitting to result from the random distribution of isotopes in natural silicon. Here we present a new model that explains these surprising experimental results, and discuss the implications for acceptor ground-state splittings observed in other isotopically mixed semiconductors, as well as for the acceptor ground state in semiconductor alloys. PMID- 12570638 TI - Asymmetric transport due to spin injection into a Kondo alloy. AB - Spin injection is found to have a significant effect on the transport properties of the Kondo alloy Cu(Fe). When a spin-polarized electron current flows from Co into Cu(Fe) wires through the Co/Cu(Fe) interface, the resistivity of the Cu(Fe) wire is suppressed near the interface, as distinct from the ordinary logarithmic increase in the resistivity at low temperatures. For the opposite current direction, no significant changes are observed. The asymmetry of the resistivity with respect to the current direction decays with a characteristic length of 1.5+/-0.4 microm at 2.5 K as the distance from the interface is increased. Possible mechanisms for the asymmetry are discussed. PMID- 12570639 TI - Fractional quantum Hall effect of composite fermions. AB - In a GaAs/AlGaAs quantum well of density 1 x 10(11) cm(-2) we observed a fractional quantum Hall effect (FQHE) at nu = 4/11 and 5/13, and weaker states at nu = 6/17, 4/13, 5/17, and 7/11. These sequences of fractions do not fit into the standard series of integral quantum Hall effects of composite fermions (CF) at nu = p/(2mp +/- 1). They rather can be regarded as the FQHE of CFs attesting to residual interactions between these composite particles. In tilted magnetic fields the nu = 4/11 state remains unchanged, strongly suggesting it to be spin polarized. The weak nu = 7/11 state vanishes quickly with tilt. PMID- 12570640 TI - Monte Carlo evaluation of non-Abelian statistics. AB - We develop a general framework to (numerically) study adiabatic braiding of quasiholes in fractional quantum Hall systems. Specifically, we investigate the Moore-Read (MR) state at nu=1/2 filling factor, a known candidate for non-Abelian statistics, which appears to actually occur in nature. The non-Abelian statistics of MR quasiholes is demonstrated explicitly for the first time, confirming the results predicted by conformal field theories. PMID- 12570641 TI - Quantum orders in an exact soluble model. AB - We find all the exact eigenstates and eigenvalues of a spin-1/2 model on square lattice: H=16g Sum S(y)(i)S(x)(i + empty set x)S(y)(i + empty set x + empty set y)S(x)(i + empty set y). We show that the ground states for g < 0 and g > 0 have different quantum orders described by Z2A and Z2B projective symmetry groups. The phase transition at g = 0 represents a new kind of phase transition that changes quantum orders but not symmetry. Both the Z2A and Z2B states contain Z2 lattice gauge theories at low energies. They have robust topologically degenerate ground states and gapless edge excitations. PMID- 12570642 TI - Phase coherent precessional magnetization reversal in microscopic spin valve elements. AB - We evidence multiple coherent precessional magnetization reversal in microscopic spin valves. Stable, reversible, and highly efficient magnetization switching is triggered by transverse field pulses as short as 140 ps with energies down to 15 pJ. At high fields a phase coherent reversal is found revealing periodic transitions from switching to nonswitching under variation of pulse parameters. At the low field limit the existence of a relaxation dominated regime is established allowing switching by pulse amplitudes below the quasistatic switching threshold. PMID- 12570643 TI - Observation of minority spin character of the new electron doped manganite La0.7Ce0.3MnO3 from tunneling magnetoresistance. AB - We report the magnetotransport characteristics of a trilayer ferromagnetic tunnel junction built of an electron doped manganite (La0.7Ce0.3MnO3) and a hole doped manganite (La0.7Ca0.3MnO3). At low temperatures the junction exhibits a large positive tunneling magnetoresistance (TMR), irrespective of the bias voltage. At intermediate temperatures below T(C) the sign of the TMR is dependent on the bias voltage across the junction. The magnetoresistive characteristics of the junction strongly suggest that La0.7Ce0.3MnO3 is a minority spin carrier ferromagnet with a high degree of spin polarization, i.e., a transport half-metal. PMID- 12570644 TI - Microscopic description of aging dynamics: fluctuation-dissipation relations, effective temperature, and heterogeneities. AB - We consider the dynamics of a diluted mean-field spin glass model in the aging regime. The model presents a particularly rich heterogeneous behavior. In order to catch this behavior, we perform a spin-by-spin analysis for a given disorder realization. We confirm the connection between statics and dynamics at the level of single degrees of freedom. Moreover, working with single-site quantities, we can introduce a new response-vs-correlation plot, which clearly shows how heterogeneous degrees of freedom undergo coherent structural rearrangements. We discuss the general scenario which emerges from our work and (possibly) applies to more realistic glassy models. Interestingly enough, some features of this scenario can be understood recurring to thermometric considerations. PMID- 12570645 TI - Quasiballistic magnetization reversal. AB - We demonstrate a quasiballistic switching of the magnetization in a microscopic magnetoresistive memory cell. By means of time resolved magnetotransport, we follow the large angle precession of the free layer magnetization of a spin valve cell upon application of transverse magnetic field pulses. Stopping the field pulse after a 180 degrees precession rotation leads to magnetization reversal with reversal times as short as 165 ps. This switching mode represents the fundamental ultrafast limit of field induced magnetization reversal. PMID- 12570646 TI - Interplay between anisotropic strain relaxation and uniaxial interface magnetic anisotropy in epitaxial Fe films on (001) GaAs. AB - Grazing incidence x-ray diffraction study of Fe epitaxial ultrathin films (1.5-13 nm) on GaAs (001) reveals an anisotropy of both domain shape and strain, with [110] and [1-10] as the principal directions. It is shown that the observed thickness-dependent strain anisotropy, together with a uniaxial interface term, can provide an unambiguous explanation to the usual in-plane magnetic anisotropy and its thickness dependence observed in this magnetic thin-film system. PMID- 12570647 TI - Long-range ferromagnetic dipolar ordering of high-spin molecular clusters. AB - We report the first example of a transition to long-range magnetic order in a purely dipolarly interacting molecular magnet. For the magnetic cluster compound Mn6O4Br4(Et2dbm)6, the anisotropy experienced by the total spin S = 12 of each cluster is so small that spin-lattice relaxation remains fast down to the lowest temperatures, thus enabling dipolar order to occur within experimental times at T(c) = 0.16 K. In high magnetic fields, the relaxation rate becomes drastically reduced and the interplay between nuclear- and electron-spin lattice relaxation is revealed. PMID- 12570648 TI - Ferromagnetism induced by clustered Co in Co-doped anatase TiO2 thin films. AB - We investigated ferromagnetism of a newly discovered ferromagnetic semiconductor Co-doped anatase TiO2 thin film, using the magnetic circular dichroism (MCD) at the Co L(2,3) absorption edges. The magnetic moment was observed to be approximately 0.1 micro(B)/Co in the measurements, but the MCD spectral line shape is nearly identical to that of Co metal, showing that the ferromagnetism is induced by a small amount of clustered Co. With thermal treatments at approximately 400 degrees C, the MCD signal increases, and the moment reaches up to approximately 1.55 micro(B)/Co, which is approximately 90% of the moment in Co metal. In the latter case, the cluster size was observed to be 20-60 nm. PMID- 12570649 TI - Optical properties of ultrathin films: evidence for a dielectric anomaly at the insulator-to-metal transition. AB - Optical properties of ultrathin layers (<50 A) of Au and Pb quench condensed on amorphous germanium (alpha-Ge) have been measured in situ at 10 K. The development of these films from an insulating state to a metallic state is traced as a function of the film thickness as well as the sheet resistance, R(square). Of particular interest is the regime of R(square) near 3000 Omega where there is an anomaly in the optical transmission. This anomaly is due to a singularity in the dielectric function when the system undergoes an electronic percolation or insulator-to-metal (I/M) transition. PMID- 12570650 TI - Minimal model for genome evolution and growth. AB - Textual analysis of typical microbial genomes reveals that they have the statistical characteristics of a DNA sequence of a much shorter length. This peculiar property supports an evolutionary model in which a genome evolves by random mutation but primarily grows by random segmental duplication. That genomes grew mostly by duplication is consistent with the observation that repeat sequences in all genomes are widespread and intragenomic and intergenomic homologous genes are preponderant across all life forms. PMID- 12570651 TI - Elastic response of single DNA molecules exhibits a reentrant collapsing transition. AB - We measured the elastic response of single DNA molecules at various concentrations of the trivalent cation, spermidine. When added spermidine caused the DNA to collapse, the force-extension curves showed either plateaus or stick release patterns depending on the concentration. The periodic stick-release response determines a characteristic length, which may reflect toroidal supercoiling. At high concentrations of spermidine, we observed the reelongation of single molecules of collapsed DNA. Thus condensation occurs between lower and upper critical concentrations, verifying that the transition is reentrant as theoretically predicted. PMID- 12570652 TI - A scheme for electrical detection of single-electron spin resonance. AB - We study a scheme for electrical detection of the spin resonance of a single electron trapped near a field effect transistor (FET) conduction channel. In this scheme, the resonant Rabi oscillations of the trapped electron spin cause a modification of the average charge of a shallow trap, which can be detected through the change in the FET channel resistivity. We show that the dependence of the channel resistivity on the frequency of the rf field can have either peak or dip at the Larmor frequency of the electron spin in the trap. PMID- 12570653 TI - Gentle dragging of reaction waves. AB - Using a recently realized "addressable catalyst surface" [Science 294, 134 (2001)]] we study the interaction of chemical reaction waves with prescribed spatiotemporal fields. In particular, we study how a traveling chemical pulse is "dragged" by a localized, moving temperature heterogeneity as a function of its intensity and speed. The acceleration and eventual "detachment" of the wave from the heterogeneity is also explored through simulation and stability analysis. PMID- 12570654 TI - Liquid crystal formation in suspensions of hard rodlike colloidal particles: direct observation of particle arrangement and self-ordering behavior. AB - We successfully prepared monodisperse, hard rodlike colloidal particles with a wide range of length-to-width ratios (L/W). In their suspensions liquid crystals, or nematic (N) and smectic (Sm) phases, spontaneously appeared. The size of the particles made it possible to directly observe their arrangement and dynamics with an optical microscope. The phase behavior observed exhibited an I (isotropic)-Sm transition for L/W=3.5-8.0 and I-N-Sm transitions for L/W=10-35. In pre-Sm transition regions, lateral clustering of the particles and subsequent layering of the clusters were observed exactly. PMID- 12570655 TI - Rotational diffusion microrheology. AB - Examining the rotational diffusion of a microparticle suspended in a soft material opens up exciting new opportunities for locally probing the frequency dependent linear viscoelastic shear modulus, G*(omega). We study the one dimensional rotational diffusion of a wax microdisk in an aqueous polymer entanglement network using light streak tracking. By measuring the disk's time dependent mean square angular displacement, , we predict the polymer solution's G*(omega) using a rotational generalized Stokes-Einstein relation. The good agreement of the predicted modulus with mechanical measurements confirms this new microrheological approach. PMID- 12570656 TI - Kinetic potential and barrier crossing: a model for warm cloud drizzle formation. AB - The kinetic potential of nucleation theory is used to describe droplet growth processes in a cloud. Drizzle formation is identified as a statistical barrier crossing phenomenon that transforms cloud droplets to drizzle size with a rate dependent on turbulent diffusion, droplet collection, and size distribution. Steady-state and transient drizzle rates are calculated for typical cloud conditions. We find drizzle more likely under transient conditions. The model quantifies an important indirect effect of aerosols on climate-drizzle suppression in clouds of higher droplet concentration. PMID- 12570657 TI - Paclitaxel resistance: molecular mechanisms and pharmacologic manipulation. AB - It has been approximately ten years since the Food and Drug Administration (FDA) approved paclitaxel for the treatment of platinum resistant epithelial ovarian carcinoma. Since the approval, the drug has found therapeutic applications in a variety of schedules and in a wide variety of epithelial malignancies. Its novel mechanism of action provided the hope that it would demonstrate anti-neoplastic activity in multidrug resistant tumor cells. Unfortunately, as with other chemotherapeutic drugs, resistance is commonly seen. Laboratory investigation has defined a wide variety of resistance mechanisms including overexpression of multidrug resistance (MDR-1) gene, molecular changes in the target molecule (betatubulin), changes in apoptotic regulatory and mitosis checkpoint proteins, and more recently changes in lipid composition and potentially the overexpression of interleukin 6 (IL-6). This review describes the in vitro molecular data that define and support the various mechanisms of resistance and critically evaluates the evidence for the participation of these mechanisms in clinically relevant paclitaxel resistance. This review also explores pharmacologic attempts to modulate paclitaxel resistance, principally through inhibition of the MDR-1 drug efflux pump. Future avenues for drug resistance research and its pharmacologic manipulation in the clinic are discussed. PMID- 12570658 TI - Mechanisms controlling sensitivity to platinum complexes: role of p53 and DNA mismatch repair. AB - Although cisplatin is effective in the treatment of different types of tumors, resistance to treatment is a major limitation. In an attempt of overcoming resistance mechanisms, a large effort has been made to generate compounds with a different geometry. At present, the most clinically relevant compounds include mononuclear (i.e. oxaliplatin) as well as multinuclear platinum complexes (i.e. BBR 3464). The mechanisms of cellular response to platinum complexes have not been completely elucidated. Among the main pathways affecting cell sensitivity of these drugs a role for p53 has been proposed at least for cisplatin and BBR 3464. Our results indicate that, also in the case of oxaliplatin, cytotoxicity is modulated by this pathway. Indeed, the effect of oxaliplatin could be reduced in tumor cells expressing mutant p53. The DNA mismatch repair system also appears to be critical in regulating cellular sensitivity to cisplatin because the loss of DNA mismatch repair results in low level of resistance to cisplatin, but not to oxaliplatin. Thus, platinum compounds are endowed with differential capability to activate pathways of p53-dependent or independent apoptosis, and differential recognition by specific cellular systems is likely to be the critical determinant of the cell fate (death/survival) after drug exposure. Further molecular studies are required to better define the precise contribution of such pathways to the cellular responses of the clinically relevant platinum complexes. A complete understanding of the molecular basis of sensitivity to platinum drugs is expected to provide useful insights for the optimization of tumor treatment. PMID- 12570659 TI - Oncogenic signaling pathways activated by RON receptor tyrosine kinase. AB - RON (Receptuer d'Origine Nantaise) is a member of the MET receptor tyrosine kinase family. RON is expressed in various cell types including macrophages, epithelial and hematopoietic cells. Its ligand, macrophage stimulating protein (MSP, also known as hepatocyte growth factor-like protein), is a multifunctional factor regulating cell growth and survival, adhesion and motility, cytokine production and phagocytosis. Accumulated data indicate that in addition to the regulation of normal cell functions, RON can be involved in cancer development and progression: (i). RON is overexpressed and constitutively active in some primary tumors and tumor cell lines; (ii). experimental mutations of RON cause oncogenic cell transformation, and (iii). RON mediates susceptibility to Friend virus-induced erythroleukemia in mice. Constitutive activation of intracellular signaling pathways such as the PI-3 kinase/AKT, beta-catenin, MAPK and JNK pathways may underlie the molecular mechanism of RON-mediated oncogenic cell transformation. The present review describes RON-activated signaling pathways, which may play an important role in tumor formation and metastasis. PMID- 12570660 TI - Molecular targets of the oncogenic transcription factor jun. AB - The Jun oncoprotein is a major component of the transcription factor complex AP 1, which regulates the expression of multiple genes essential for cell proliferation, differentiation and apoptosis. Constitutive activation of endogenous AP-1 is required for tumor formation in avian and mammalian cell transformation systems, and also occurs in distinct human tumor cells suggesting that AP-1 plays an important role in human oncogenesis. The highly oncogenic v jun allele capable of inducing neoplastic transformation in avian fibroblasts and fibrosarcomas in chicken as a single oncogenic event, was generated by mutation of the cellular c-jun gene during retroviral transduction. Hence, avian cells represent an excellent model system to investigate molecular mechanisms underlying jun-induced cell transformation. Approaches aimed at the identification of genes specifically deregulated in jun-transformed fibroblasts have led to the identification of several genes targeted by oncogenic Jun. Some of the activated genes represent direct transcriptional targets of Jun encoding proteins, which are presumably involved in cell growth and differentiation. Genes suppressed in v-jun-transformed cells include several extracellular proteins like components of the extracellular matrix or proteins involved in extracellular signalling. Due to aberrant regulation of multiple genes by the Jun oncoprotein, it is assumed that only the combined differential expression of Jun target genes or of a subset thereof contributes to the conversion of a normal fibroblast into a tumor cell displaying a phenotype typical of jun-induced cell transformation. It has already been shown that distinct activated targets exhibit partial transforming activity upon over-expression in avian fibroblasts. Also, distinct target genes silenced by v-Jun inhibit tumor formation when re-expressed in v-jun transformed cells. The protein products of these transformation-relevant genes may thus represent potential drug targets for interference with jun-induced tumorigenesis. PMID- 12570661 TI - The prodigiosins: a new family of anticancer drugs. AB - Apoptosis is involved in the action of several (and perhaps all) cancer chemotherapeutic agents. Prodigiosins, a family of natural red pigments characterized by a common pyrrolylpyrromethene skeleton, are produced by various bacteria. Three members of the prodigiosin family, viz. prodigiosin (PG), undecylprodigiosin (UP) and cycloprodigiosin hydrochloride (cPrG.HCl), have immunosuppressive properties and apoptotic effects on cancer cells in vitro and in vivo. Their cytotoxic effect is attributed to the presence of the C-6 methoxy substituent. The A-pyrrole ring plays a key role in both the copper nuclease activity and the cytotoxicity of prodigiosins. Here, we have reviewed the pharmacological activity of PG and related compounds, including novel synthetic PG-derivatives with lower toxicity. The mechanism of action for these molecules is a current topic in biomedicine. The molecular targets of prodigiosins are also discussed. PMID- 12570662 TI - Garlic [Allium sativum]: a review of its potential use as an anti-cancer agent. AB - Garlic [Allium sativum] is among the oldest of all cultivated plants. It has been used as a medicinal agent for thousands of years. It is a remarkable plant, which has multiple beneficial effects such as antimicrobial, antithrombotic, hypolipidemic, antiarthritic, hypoglycemic and antitumor activity. In this review, we will discuss particularly the largely preclinical use of this agent in the treatment and prevention of cancer. A number of studies have demonstrated the chemopreventive activity of garlic by using different garlic preparations including fresh garlic extract, aged garlic, garlic oil and a number of organosulfur compounds derived from garlic. The chemopreventive activity has been attributed to the presence of organosulfur compounds in garlic. How this is achieved is not fully understood, but several modes of action have been proposed. These include its effect on drug metabolizing enzymes, antioxidant properties and tumor growth inhibition. Most of these studies were carried out in the animal models. Also, recent research has focused on the antimutagenic activity of garlic. Recently, it has been observed that aged garlic extract, but not the fresh garlic extract, exhibited radical scavenging activity. The two major compounds in aged garlic, S-allylcysteine and S-allylmercapto-L-cysteine, had the highest radical scavenging activity. In addition, some organosulfur compounds derived from garlic, including S-allylcysteine, have been found to retard the growth of chemically induced and transplantable tumors in several animal models. Therefore, the consumption of garlic may provide some kind of protection from cancer development. PMID- 12570663 TI - Comparative immunology. AB - Comparative Immunology has gained wide acceptance in biology, as an offspring of immunology and an amalgam of immunology and zoology. The prescient experiments of Metchnikoff on phagocytosis in invertebrates during the 19th century served to splinter immunology into its two main components: cellular and humoral. There is much interest in the immune system of invertebrates as representing early models or precursors of the innate system of vertebrates that by contrast possess the innate system as well as the more highly evolved adaptive system. With respect to mechanisms, we think of the invertebrate system as innate, natural, non-specific, non-anticipatory, and non-clonal. Innate immunity operates through leukocytes that are not components of the macrophage T and B interrelationships that characterize vertebrate adaptive immunity that is adaptive, induced, specific, anticipatory, and clonal. This symposium on invertebrate immunology has provided an overview of what is current and crucial to understanding the larger field of comparative immunology. Comparative immunology is now an established field, here since Metchnikoff but officially since about 1977, with a journal (Developmental and Comparative Immunology) (DCI) and an International Society of Developmental and Comparative Immunology (ISDCI). During this short but vigorous history several national, adherent societies have been organized in Japan, Italy and Germany with sporadic interest in a national group in the USA. Nevertheless, comparative immunology is here as vital to zoology in general and to immunology in particular as we delve deeper into unique but also shared characteristics. PMID- 12570664 TI - Leeches: immune response, angiogenesis and biomedical applications. AB - The innate immune response is the first line of defence strategies in invertebrates against attack of infectious agents. A detailed analysis of the immune mechanisms involved in annelids has been performed in oligochaets, but few data are available in polichaets and hirudineans. The aim of this review is to describe the responses of leeches to different kinds of stimuli (infections following non-self agent attacks, surgical lesions, grafts). Furthermore, the use of this invertebrate as a novel experimental model to be used to screen drugs and genes, which are responsible for positive and negative modulation of angiogenesis, is discussed. PMID- 12570665 TI - Annelid neuroimmune system. AB - Neuropeptides have been found in nervous central or immune systems of Annelids. Since these signaling molecules are found free in the hemolymph, they are considered as hormones. Hormonal processes along with enzymatic processing similar to that found in vertebrates occur in annelids. Furthermore, amino acid sequence determination of annelids precursor gene products reveals the presence of the respective peptides that exhibit high sequence identity to their mammalian counterparts. Nevertheless, specific neuropeptides to annelids or invertebrates have also been in these animals. These peptides are flanked by potential proteolytic signal sites for the various known enzymes confirming that annelids neuropeptide precursors are processed in a similar manner to that described in mammals i.e. implicating prohormone convertase enzymes. PMID- 12570666 TI - Antimicrobial compounds of low molecular mass are constitutively present in insects: characterisation of beta-alanyl-tyrosine. AB - The number of bacterial and fungal strains that have developed resistance against the classical antibiotics continues to grow. The intensified search for new antibiotic lead compounds has resulted in the discovery of numerous endogenous peptides with antimicrobial properties in plants, bacteria and animals. Their possible applications as anti-infective agents are often limited by their size, in reference to production costs and susceptibility to proteases. In this article, we report recent isolations of antimicrobial compounds from insects, with molecular masses less than 1 kDa. Experimental approaches are discussed and the first data on the antimicrobial properties of beta-alanyl-tyrosine (252 Da), one of such low molecular mass compounds isolated from the fleshfly Neobellieria bullata, are presented. We also offer evidence for the constitutive presence of antimicrobial compounds in insects of different orders, in addition to the previously identified inducible antimicrobial peptides. PMID- 12570667 TI - Probiotics as biotherapeutic agents: present knowledge and future prospects. AB - Since the early observations of Elie Metchnikoff, a wealth of experiments have described the use of selected microorganisms, mainly belonging to the lactic acid bacteria family, for the prevention or treatment of a variety of pathological situations. Nevertheless, the mechanisms underlying the proposed actions remain vastly unknown, partly as a consequence of the complexity of the gastro intestinal ecosystem with which these biotherapeutic agents are expected to interact, but also because of the increasing variety of strains considered to have potential probiotic characteristics. During the past decades, however, the beneficial effect of specific strains in preventing or treating intestinal disorders has been substantiated by well-controlled clinical trials. Increasing evidence, including human studies, is also supporting the immunomodulatory role attributed to given lactic acid bacterial strains. The desire by consumers to use natural methods for health maintenance rather than long-term chemotherapeutic agents (i.e. antibiotics), linked to their expectation that food becomes a source of prolonged well-being, supports the speculation that the probiotic market will expand rapidly. Much of this growth will also depend on the reliability of claims that these products will bare. Therefore, the legislator will have to provide clear rules and regulations which will depend on measurable biomarkers and criteria based on scientific evidence. These commercial and legislative needs will hopefully provide scientists with the resources necessary to conduct the multidisciplinary research required to establish facts and mechanisms of action for carefully selected probiotic strains. These research results will probably be as essential for the positioning of probiotic preparations as either a food, a food supplement or as pharmaceutical preparation. PMID- 12570668 TI - Study of atrazine effects on Pacific oyster, Crassostrea gigas, haemocytes. AB - Shellfish farming is an important economic activity around the world. This activity often takes place in areas subjected to various recurring pollutions. The recrudescent use of herbicides in agriculture including atrazine implies pollutant transfer towards aquatic environment in estuarine areas. Harmful effects of such substances on animals in marine environment, particularly on cultured bivalves, are poorly documented. Bivalve molluscs such as mussels and oysters have been postulated as ideal indicator organisms because of their way of life. They filter large volumes of seawater and may therefore accumulate and concentrate contaminants within their tissues. Moreover, development of techniques allowing effect analysis of such compounds on bivalve biology may lead to the development of diagnosis tools adapted to analyze pollutant transfer towards estuarine areas. In this context, influence of atrazine on defence mechanisms was analyzed in Pacific oysters, Crassostrea gigas. Atrazine was tested in vitro and in vivo on oyster haemocytes, and its effects were analyzed by flow cytometry. Haemocyte viability, cell cycle and cellular activities were monitored. Atrazine induced no significant effect in oyster under tested conditions except for peroxidase activity. PMID- 12570669 TI - Intestinal epithelial toll-like receptors: to protect. And serve? AB - The innate immune system uses a series of pattern recognition receptors to detect the presence of pathogens thus allowing for rapid host defense responses to invading microbes. A key component of such receptors are the "toll-like receptors" (TLRs), which recognize a panel of microbial molecules that tend to be somewhat invariant, at least in select regions, thus permitting a relatively small number of receptors to recognize a large number of different microbes. Accordingly, this panel of TLRs bears little ability to distinguish between commensal and pathogenic microbes as such organisms generally bear far more structural similarities than differences between them. For the professional phagocytic cells classically considered to be the primary mediators of innate immunity such distinction between commensal and pathogenic microbes is not particularly important since any microbe that breaches the outer host defensive barriers to reach these phagocytes, whether doing so by a pathogen-specific or opportunistic mechanism, is likely potentially hazardous to its host. However, epithelial cells that line mucosal surfaces, thus being on the front line of host defense, also play an active role in innate immunity particularly by secreting chemokines and other immune mediators in response to pathogenic microbes. Epithelial cells have been reported to express several TLRs suggesting these receptors play a role in intestinal epithelial innate immune signaling pathways. However, since some mucosal surfaces such as the intestinal epithelium are normally densely colonized by a wide variety of microbes, the ability to distinguish the occasional pathogen from the sea of commensals presents an important challenge. This minireview considers the current findings regarding TLR expression in the intestinal epithelium and the role these receptors might serve in host defense. PMID- 12570670 TI - Role of DAP12 in innate and adaptive immune responses. AB - DAP12 is a novel immunoreceptor tyrosine-based activation motifs (ITAM)-bearing transmembrane adapter molecule. This molecule, together with its partner receptor complex molecules including the killer cell activating receptors (KARs), myeloid DAP12 associating lectin-1 (MDL-1), triggering receptor expressed on myeloid cells 1/2/3 (TREM-1, TREM-2, TREM-3), and signal regulatory protein beta1 (SIRPbeta1), are expressed on the surface of NK and myeloid cells including antigen presenting cells. While the function of DAP12 and its associating molecules has just begun to be unveiled, emerging evidence suggests that these molecules play an important role in both innate and adaptive immune responses. In this review, we intend to provide an overview on what have been known and are still unknown to date about the function of these molecules based on the observations made by us and others. PMID- 12570671 TI - Mast cell cytokine and chemokine responses to bacterial and viral infection. AB - Mast cells have been most widely studied in the context of allergic disease but also play a critical role in host defence against bacterial infection, most elegantly demonstrated in studies using mast cell deficient w/wv mice. There is less data available concerning the role of mast cells in defence against viral pathogens, however, mast cells have been demonstrated to be a potential reservoir of infection for several pathogens, such as HIV-1 and dengue, and capable of producing mediators following challenge with a number of viral products. Traditional mast cell mediators such as histamine, protease enzymes and leukotrienes are important for effective host responses. The cytokines and chemokines produced by mast cells in response to pathogens are known to profoundly alter the nature of the innate immune response and its effectiveness in eliminating infection. Cytokine and chemokine production by mast cells is closely regulated and may occur independently of classical mast cell degranulation. Depending upon the nature of the stimulus or type of infection, a unique profile of cytokines is induced. In this review, we will examine the role and regulation of mast cell cytokines and chemokines in the context of a number of bacterial and viral infections, emphasizing the multiple receptor mechanisms used to activate mast cells. This area of research is still in its early stages and much work remains to be done. However, understanding the unique properties of resident tissue mast cells and how their cytokine responses are regulated by pathogens or pathogen products, will provide important opportunities for the therapeutic manipulation of local immune responses. PMID- 12570672 TI - Cytokines in the pathogenesis of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a common cause of morbidity and mortality. The term is heterogenous and encompasses a number of distinct but often overlapping phenotypes including chronic bronchitis, small airways obstruction, emphysema and in some individuals, a systemic component. Although there have been significant advances in understanding the pathophysiology of COPD, understanding of the role of the inflammation in the pathogenesis of the condition remains in its infancy. Indeed, cytokines that are known to orchestrate the inflammatory response in asthma and other inflammatory diseases are only beginning to be reported in COPD. In this review, we highlight the potential role of cytokines in the development of mucus hypersecretion observed in chronic bronchitis and the morphological changes observed in the small airways and airspaces contributing to the development of airflow limitation and respiratory failure respectively. We report evidence that exacerbations are linked to increased expression of pro-inflammatory cytokines and that the wasting and skeletal muscle dysfunction observed in some patients is most probably related to the presence of a systemic inflammatory response. In addition transgenic and gene therapy technology has been used to explore the temporal and co-ordinated role of cytokines in the development of COPD animal models. Enhanced understanding of the events involved in the pathogenesis of COPD will lead to the development of therapy with potential to modify the observed progressive decline in lung function and impact on the development of the illness. PMID- 12570673 TI - Re-evaluation of fibrogenic cytokines in lung fibrosis. AB - Idiopathic Pulmonary Fibrosis (IPF) is a chronic interstitial lung disease which results in end-stage fibrosis. The pathogenesis is believed to be related to a dysregulation in cross-talk between inflammatory and structural cells, mediated by various cytokines, chemokines and growth factors, which are responsible for the maintenance of tissue homeostasis and which coordinate the response to injury. The large number of mediators involved and the complexity of their interaction makes it difficult to identify the factors responsible for initiation of fibrogenesis and progression to chronicity. Whether a mediator's presence in fibrotic lung is as a result of tissue injury or if it playsan active role in disease onset and progression has been partly answered by the use of transient and/or permanent transgenic and gene knock-out approaches to over-express single factors at a time. Chemokines such as interleukin-8 (IL-8), RANTES, IP-10, MIG or lymphotactin, do not appear to induce fibrosis when over-expressed in rodent lung. Amongst many tested, four cytokines and growth factors have been found to be pro-fibrotic; IL-1beta, which demonstrates marked inflammation, tissue damage and chronic fibrosis, TNF-alpha, which induces inflammation and mild fibrosis, and GM-CSF, which induces moderate inflammation and fibrosis. A common finding with these cytokines are increased lung TGF-beta levels, proportionate to the degree of fibrosis generated, while TGF-beta itself causes minor inflammation but marked progressive chronic fibrosis. A growth factor 'downstream' from the pro fibrotic effects of TGF-beta, CTGF, is a likely critical mediator. However, over expression of CTGF produces only mild and reversible fibrosis. PMID- 12570674 TI - Role of cytokines in the development and maintenance of memory T cells during respiratory viral infection. AB - Much progress has been made in understanding the relationship between cytokines, T cell development, and the maintenance of memory T cells by examining the immune response to respiratory viral infections. Most of these studies have examined the T cell response to viruses that cause acute infection of limited duration, and have focused on the interplay between cytokines and individual responses by T cell subsets. This reductionism approach has been useful to piece together the puzzle of the host-immune response to respiratory virus infection, and has added to the holistic view of the networks involved in homeostatic control of T cell development and maintenance. This review addresses aspects of T cell biology that constitute the response to respiratory viral infections. PMID- 12570675 TI - Role of type I cytokines in host defense against Mycobacterium avium infection. AB - Mycobacterium avium is a human pathogen that causes infection in immunocompetent as well as immunocompromised patients. Infection is acquired both by the respiratory and gastrointestinal routes, and bacterial invasion of mucosal epithelial cells is characteristic. M. avium crosses the mucosal barrier without triggering substantial inflammatory response. Once in the intestinal submucosa or in the alveolar space M. avium infects macrophages. Intracellular bacteria block the production of cytokines involved in the host response against the infection, such as TNF-alpha and IL-12, and suppress antigen presentation by the macrophage. Innate response against the infection is effective to certain extent but the ability of the bacterium to remain "silent" for a period of time prevents neutrophil and NK cells from effectively controlling the establishing of the infection. CD4+ T cells as well as CD8+ T cells are activated, although only CD4+ T cells appear to be effective in inducing anti-M. avium activity in macrophages. M. avium-specific CD8+ T cells undergo apoptosis early in the infection. Therefore, the immune mechanisms of the host and bacterial strategies for survival are complex and fascinating. PMID- 12570676 TI - Role of cytokines in Chlamydia trachomatis protective immunity and immunopathology. AB - Diseases caused by Chlamydia trachomatis infection and its sequelae represent major public health concerns worldwide. In order to rationally develop an effective vaccine to chlamydial infection, we need a better understanding of the mechanisms underlying the protective and pathological immune responses to chlamydial antigens. Recent studies in chlamydial immunobiology have demonstrated a close link between cytokine production patterns and the type of immune responses to this pathogen. In particular, IL-10 has been found to be associated with susceptibility to chlamydial infection and the typical pathological changes caused by the infection such as granuloma formation and fibrosis. Although Th1 type delayed hypersensitivity (DTH) is associated with protective immunity, the Th2 type DTH documented in interferon (IFN)-gamma gene knockout (KO) mice fails to control chlamydial infection. The Th2 type DTH is characterized by eosinophil and neutrophil infiltration and is associated with high levels of IL-4 and IL-5 production. The capability of dendritic cell (DC) in initiating T cell response to Chlamydia has been shown in studies using cultured DCs and a DC line. Further studies should be performed to elucidate the role played by DCs in natural chlamydial infection and the potential involvement of DC subsets in directing immune responses to chlamydial infection. PMID- 12570677 TI - Cytokine therapeutics for the treatment of sepsis: why has nothing worked? AB - Several clinical trials have attempted to treat sepsis by blocking certain aspects of the inflammatory response. Tumor necrosis factor and interleukin 1 have been specific targets for inhibition but none of the trials have been successful. These trials were started on the basis of preclinical trials that suggested these would be effective. There were three lines of evidence to support the idea of cytokine inhibition. First, patients with increased levels of cytokines are more likely to die. Second, experimental animal models demonstrated that blocking the cytokines would improve outcome. Third, injection of purified, recombinant cytokines would cause both organ injury and death in experimental animals. Several additional aspects of the inflammatory response have been discovered since these trials were initiated. Included among these potential new targets are interleukin 18 and HMG-1. However, before new clinical trials are started there must be careful consideration of why previous interventions were not effective. The concept of blocking a single elevated cytokine may be too simple to deal with the complex problem of sepsis. As patients move through different phases of the septic response, there may be intervals when it is appropriate to inhibit multiple cytokines while at other times it may be appropriate to augment the immune response. PMID- 12570678 TI - Immune modulation by plasmid DNA-mediated cytokine gene transfer. AB - Autoimmune diseases frequently develop as a result of an abnormal activation of autoreactive T cells, excessive production of proinflammatory cytokines, particularly by CD4(+) Th1 cells, and subsequent tissue destruction. Cytokine dependent immunotherapy can be applied to alter the balance between Th1 and Th2 cell activity, or proinflammatory versus immunosuppressive cytokine profiles. Cytotoxic T lymphocyte (CTL) and/or macrophage activity can also be suppressed. Gene transfer offers numerous advantages for the in vivo delivery of cytokines or their receptors for immunotherapeutic use. We have relied on the injection of naked plasmid DNA into skeletal muscle to deliver therapeutic genes. In particular, we have successfully used this approach to deliver neutralizing cytokine receptors such as interferon gamma (IFNgamma)-receptor-Ig fusion proteins or anti-inflammatory cytokines such as transforming growth factor beta-1 (TGF-beta1) and interleukin 4 (IL-4). Intramuscular gene therapy is effective in protecting against several experimental autoimmune diseases including insulin dependent diabetes mellitus (IDDM), experimental allergic encephalomyelitis (EAE), and systemic lupus erythematosus (SLE). Another promising approach involves DNA vaccination by plasmid-based codelivery of genes encoding an autoantigen and either a cytokine or other immunomodulatory molecule. Plasmid vectors offer interesting advantages over viral vectors, since they are simple to produce, non-immunogenic and non-pathogenic. They can be repeatedly administered with relatively prolonged periods of expression in vivo, ranging from weeks to months after each injection. Plasmid-based intramuscular gene transfer has great therapeutic potential in the areas of autoimmune and inflammatory disorders. PMID- 12570679 TI - Receptor-based design of cytokine therapeutics. AB - Cytokines hold huge potential for the treatment of disease due to their often fundamental roles in development and homeostasis. However, it is this same primary biological function that can both cause disease through dysregulation as well as prevent their therapeutic use due to systemic consequences arising from this inherent pleiotropy. Molecularly, this can be explained through an understanding of the receptor system specific to each cytokine and the cells on which they are expressed. This knowledge has been exploited to yield muteins (mutated proteins) that exhibit selective, and sometimes novel, biological properties dependent upon receptor subunit usage. In some cases, these muteins have been evaluated in clinical trials and have been approved for clinical use; in most instances, however, these muteins are not suitable for therapeutic application due to intrinsic characteristics of the muteins themselves or the cellular and receptor system to which they are directed. Ultimately, molecular insight to the biological processes governing disease pathology underlies the successful application of mutein-based therapy. The clinical success enjoyed by a subset of these proteins signals the advent of a new mode of therapeutic protein development. PMID- 12570680 TI - Inhibiting cyclin-dependent kinase/cyclin activity for the treatment of cancer and cardiovascular disease. AB - Excessive cell proliferation contributes to the pathobiology of human diseases with a high health and socio-economic impact, including cancer and vascular occlusive diseases (e. g., atherosclerosis, in-stent restenosis, transplant vasculopathy, and vessel bypass graft failure). Recent advances in the understanding of the molecular networks governing the hyperplastic growth of tumors and vascular obstructive neointimal lesions have provided new perspectives for preventive and therapeutic strategies against these disorders. Mammalian cell proliferation requires the activation of several cyclin-dependent protein kinases (CDKs). Postranslational activation of CDKs is a complex process that involves their association with regulatory subunits called cyclins. The activity of CDK/cyclin holoenzymes is negatively regulated through their interaction with members of the CDK family of inhibitory proteins (CKIs). Moreover, over fifty low molecular weight pharmacological CDK inhibitors that target the ATP-binding pocket of the catalytic site of CDKs have been identified. In this review, we will discuss the use of pharmacological and gene therapy strategies against CDK/cyclins in animal models and clinical trials of cancer and cardiovascular disease. PMID- 12570681 TI - DAB389EGF fusion protein therapy of refractory glioblastoma multiforme. AB - Primary brain tumors including anaplastic astrocytomas and glioblastoma multiforme are difficult to treat because of their locally invasive nature and chemoradioresistance. Novel therapies are needed. One class of therapeutics is fusion proteins consisting of peptide toxins fused to brain tumor selective ligands. DAB389EGF is a fusion protein composed of the catalytic and translocation domains of diphtheria toxin fused via a His-Ala linker to human epidermal growth factor (EGF). DAB389EGF is selectively toxic to EGF receptor (EGFR) overexpressing cells. Close to half of all high-grade primary brain tumors have EGFR gene amplification and EGFR overexpression. With the use of convection enhanced delivery (CED), DAB389EGF may be delivered locally at high concentrations to the brain tumor. CED would avoid many of the pharmacologic and toxicologic barriers which have limited effective use of this agent including rapid clearance from the circulation, high anti-diphtheria toxin antibody titers in the blood and toxicities to the liver and kidney. Both cell lines and animal models are available to assess the potential of this agent for brain tumor therapy. Since significant amounts of clinical grade DAB389EGF are available, some careful additional preclinical efficacy work should lead to testing of this agent in patients within the next few years. PMID- 12570682 TI - Recombinant secretory immunoglobulin A in passive immunotherapy: linking immunology and biotechnology. AB - The use of monoclonal antibodies has become routine in the research and diagnostic laboratories, but the potential of antibody molecules in public health and medical applications is still far from its maximum. Most infections begin at mucosal surfaces, and this is certainly not only a stroke of good fortune if mother's milk serves as a natural delivery vehicle for antibodies protecting the gastrointestinal tract of nursing infants. Mammary gland or other mucous secretions containing numerous antibody specificities provide an efficient mean to immediately protect a mucosal surface against pathogens, which have never been encountered by the host. From a public health perspective, topical passive immunization of mucosal surfaces with monoclonal antibodies can block entry and transmission of bacteria, viruses, fungi and parasites that infect humans, and thus defeat some key immune evasion strategies designed by many pathogens. The chief antibody on most mucosal surfaces is secretory immunoglobulin A (SIgA), a polypeptide complex comprising dimeric IgA, the connecting J chain, and the secretory component. The molecular stability, tetravalency, and strong anti inflammatory properties make SIgA particularly well suited to fulfill the function of passive protective immunity when applied exogenously to mucosal surfaces. The review will give an overview of the basic concepts underlying mucosal immunity, present the molecular mechanisms whereby SIgA prevents mucosal infections, cover the last advances in the topic of recombinant SIgA production, and examine how structure-function relationship in SIgA will help designing molecules with novel properties for passive immunotherapy. PMID- 12570683 TI - Selecting the components for a safe and efficient tuberculosis subunit vaccine- recent progress and post-genomic insights. AB - Prophylactic vaccination against tuberculosis with BCG gained much of the credit for the decline of TB in Europe. However, with TB resurgent in many parts of the world, better vaccines are urgently needed. To improve on BCG, a rapid, rational approach to vaccine discovery is needed. Fortunately, advances in the fields of molecular biology and computer science have spawned new disciplines: Genomics, Proteomics and Transcriptomics are transforming the ways in which candidate vaccine antigens are discovered. In this review, we discuss how these new approaches have accelerated the pace of antigen discovery and vaccine development, and highlight some of the most promising new candidate vaccines and vaccine targets. PMID- 12570684 TI - Drug discovery today. AB - In recent years, tools for the development of new drugs have been dramatically improved. These include genomic and proteomic research, numerous biophysical methods, combinatorial chemistry and screening technologies. In addition, early ADMET studies are employed in order to significantly reduce the failure rate in the development of drug candidates. As a consequence, the lead finding, lead optimization and development process has gained marked enhancement in speed and efficiency. In parallel to this development, major pharma companies are increasingly outsourcing many components of drug discovery research to biotech companies. All these measures are designed to address the need for a faster time to market. New screening methodologies have contributed significantly to the efficiency of the drug discovery process. The conventional screening of single compounds or compound libraries has been dramatically accelerated by high throughput screening methods. In addition, in silico screening methods allow the evaluation of virtual compounds. A wide range of new lead finding and lead optimization opportunities result from novel screening methods by NMR, which are the topic of this review article. PMID- 12570692 TI - The nitric oxide producing reactions of hydroxyurea. AB - Hydroxyurea is used to treat a variety of cancers and sickle cell disease. Despite this widespread use, a complete mechanistic understanding of the beneficial actions of this compound remains to be understood. Hydroxyurea inhibits ribonucleotide reductase and increases the levels of fetal hemoglobin, which explains a portion of the effects of this drug. Administration of hydroxyurea to patients results in a significant increase in levels of iron nitrosyl hemoglobin, nitrite and nitrate suggesting the in vivo metabolism of hydroxyurea to nitric oxide. Formation of nitric oxide from hydroxyurea may explain a portion of the observed effects of hydroxyurea treatment. At the present, the mechanism or mechanisms of nitric oxide release, the identity of the in vivo oxidant and the site of metabolism remain to be identified. Chemical oxidation of hydroxyurea produces nitric oxide and nitroxyl, the one-electron reduced form of nitric oxide. These oxidative pathways generally proceed through the nitroxide radical (2) or C-nitrosoformamide (3). Biological oxidants, including both iron and copper containing enzymes and proteins, also convert hydroxyurea to nitric oxide or its decomposition products in vitro and these reactions also occur through these intermediates. A number of other reactions of hydroxyurea including the reaction with ribonucleotide reductase and irradiation demonstrate the potential to release nitric oxide and should be further investigated. Gaining an understanding of the metabolism of hydroxyurea to nitric oxide will provide valuable information towards the treatment of these disorders and may lead to the development of better therapeutic agents. PMID- 12570693 TI - Inhibitors of 17 beta-hydroxysteroid dehydrogenases. AB - The 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) play an important role in the regulation of steroid hormones, such as estrogens and androgens, by catalysing the reduction of 17-ketosteroids or the oxidation of 17beta hydroxysteroids using NAD(P)H or NAD(P)(+) as cofactor. The enzyme activities associated with the different 17beta-HSD isoforms are widespread in human tissues, not only in classic steroidogenic tissues, such as the testis, ovary, and placenta, but also in a large series of peripheral intracrine tissues. In the nineties, several new types of 17beta-HSD were reported, indicating that a fine regulation is carried out. More importantly, each type of 17beta-HSD has a selective substrate affinity, directional (reductive or oxidative) activity in intact cells, and a particular tissue distribution. These findings are important for understanding the mode of action of the 17beta-HSD family. From a therapeutic point of view, this means that selectivity of drug action could be achieved by targeting a particular 17beta-HSD isozyme. Consequently, each study that leads to better knowledge of the inhibition of 17beta-HSDs deserves attention from scientists working in this and related fields. Being involved in the last step of the biosynthesis of sex steroids from cholesterol, the 17beta-HSD family constitutes an interesting target for controlling the concentration of estrogens and androgens. Thus, inhibitors of 17beta-HSDs are useful tools to elucidate the role of these enzymes in particular biological systems or for a therapeutic purpose, especially to block the formation of active hydroxysteroids that stimulate estrogeno-sensitive pathologies (breast, ovarian, and endometrium cancers) and androgeno-sensitive pathologies (prostate cancer, benign prostatic hyperplasia, acne, hirsutism, etc). Few review articles have however focussed on 17beta-HSD inhibitors although this family of steroidogenic enzymes includes interesting therapeutic targets for the control of several diseases. Furthermore, inhibitors of 17beta-HSDs constitute a growing field in biomedical research and there is a need for an exhaustive review on this topic. In addition to giving an up-to-date description of inhibitors of all 17beta-HSD isoforms (types 1-8), the present review will also address, when possible, the isoform selectivity and residual estrogenic or androgenic activity often associated with steroidal inhibitors. PMID- 12570694 TI - Proteasome inhibitors as therapeutic agents: current and future strategies. AB - In cells, protein degradation is a key pathway for the destruction of abnormal or damaged proteins as well as for the elimination of proteins whose presence is no longer required. Among the various cell proteases, the proteasome, a multicatalytic macromolecular complex, is specifically required for the degradation of ubiquitinated proteins. In normal cells, the proteasome ensures the elimination of numerous proteins that play critical roles in cell functions throughout the cell cycle. Defects in the activity of this proteolytic machinery can lead to the disorders of cell function that is believed to be the root cause of certain diseases. Indeed, many proteins involved in the control of cell cycle transitions are readily destroyed by the proteasome once their tasks have been accomplished. Moreover, because proteasome inhibitors can provoke cell death, it has been suggested that proteasomes must be continually degrading certain apoptotic factors. For these reasons, proteasome inhibition has become a new and potentially significant strategy for the drug development in cancer treatment. The proteasome possesses three major peptidase activities that can individually be targeted by drugs. Different classes of proteasome inhibitors are reviewed here. In addition, we present new pseudopeptides with the enriched nitrogen backbones bearing a side chain and a modified C-terminal position that inhibit proteasome activity. PMID- 12570695 TI - Beta-propellers: associated functions and their role in human diseases. AB - The beta-propeller fold appears as a very fascinating architecture based on four stranded antiparallel and twisted beta-sheets, radially arranged around a central tunnel. Similar to the alpha/beta-barrel (TIM-barrel) fold, the beta-propeller has a wide range of different functions, and is gaining substantial attention. Some proteins containing beta-propeller domains have been implicated in the pathogenesis of a variety of diseases such as cancer, Alzheimer, Huntington, arthritis, familial hypercholesterolemia, retinitis pigmentosa, osteogenesis, hypertension, and microbial and viral infections. This article reviews some aspects of 3D structure, amino acids sequence regularities, and biological functions of the proteins containing beta-propeller domains. Major emphasis has been laid on beta-propellers whose functions are associated to human diseases. Recent research efforts reported in the fields of protein engineering, drug design, and protein structure-function relationship studies, concerning the beta propeller architecture, have also been discussed. PMID- 12570696 TI - Surrogate markers as a guide to evaluate response to antiretroviral therapy. AB - The development of an increasing number of antiretroviral agents has dramatically reduced HIV-associated morbidity and mortality. However, most of these drugs have been approved through clinical trials where only surrogate markers for clinical endpoints have been used. Ideally, a surrogate marker should be biologically plausible, predictive of disease progression and measurable by standardized assays. Historically, a number of candidate markers have been explored for monitoring the course of HIV infection and response to treatment. While the level of plasma HIV RNA and the absolute numbers of peripheral CD4+ T cells have eventually become the reference markers in clinical practice, several additional parameters are still being evaluated to improve our knowledge of the virus-host interaction, discriminate between apparently equivalent stages and further refine antiretroviral treatment. Advances in molecular methods and growing elucidation of HIV dynamics in vivo have made it possible to consider several molecular virologic parameters as candidate markers for treatment response, including intracellular levels of different HIV RNA species and amount of integrated and unintegrated HIV DNA. Much effort has been recently devoted to the definition of immunological parameters as prognostic markers. The abnormal activation induced by HIV on the immune system represents a major pathogenetic feature of HIV infection. Immune activation may be evaluated by the analysis of activation markers expressed on the cell membrane and by the quantification of soluble plasma molecules released by activated cells. Such markers of immune activation have an important prognostic significance in terms of disease progression and might be suitable for the monitoring and prognosis of antiretroviral therapies. In the late years, the possibility of extending potent antiretroviral therapies to developing countries has raised the need of simple, reliable and cost effective tests to measure prognostic markers for disease evolution and assessment of therapy efficacy. This review summarizes the benefits and limits of reference and candidate surrogate markers and their integration for optimal antiretroviral therapy. PMID- 12570697 TI - Cyclin-dependent kinase inhibitors: cancer killers to neuronal guardians. AB - The development of small molecule kinase inhibitors as potential cancer therapeutics is an area of intense interest, and a subset of these agents target cyclin-dependent kinase (CDK) activity. Ten distinct CDKs (1-9, 11), when paired with their cyclin activators, are integral to such diverse processes as cell cycle control, neuronal development, and transcriptional regulation. Mutation and/or aberrant expression of certain CDKs and their regulatory counterparts are associated with uncontrolled proliferation and tumorigenesis. As such, CDK selective inhibitors (CDKIs) that attenuate or prevent tumor growth have been developed. Recently, interest in the therapeutic potential of CDKIs has expanded to include neurodegenerative diseases, where dysregulated CDK activity has been linked to the pathogenesis of Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), and stroke. Specifically, aberrant activation of cell cycle CDKs or CDK5 is associated with apoptosis and neuronal dysfunction in response to various neuronal stressors. To date, CDKIs have shown promise as neuroprotective agents in the research laboratory and, in the future, may prove useful in the neurology clinic. PMID- 12570698 TI - Chromatographic retention parameters in medicinal chemistry and molecular pharmacology. AB - The importance of lipophilicity for pharmacological and toxicological potency of xenobiotics has been recognized for a century. The reference lipophilicity scale is defined by the logarithm of partition coefficient, log P, determined in the l octanol-water partition system. The tediousness of determinations and limited interlaboratory reproducibility of log P, on one hand, and the observations of linear relationship between log P and chromatographic retention parameters, on the other hand, gave rise to the substitution of the former by the readily available chromatographic data. Since its introduction, the reversed-phase high - performance liquid chromatography (HPLC), which has been viewed in terms of partition of a solute between a polar, aqueous mobile phase and a nonpolar stationary phase appeared especially suitable for lipophilicity (hydrophobicity) determination. The method got wide acceptance and has officially been recommended by the OECD. Fundamental relationships between chromatographic parameters are reviewed from the point of view of convenient and reliable lipophilicity measurements. The advantages and disadvantages of the stationary phase materials, which are presently employed for the determination of lipophilicity as well as those of specific HPLC systems and procedures, are critically reported. The literature on the application of chromatographic and electrochromatographic methods for assessment of lipophilicity of xenobiotics is reviewed. A separate paragraph is devoted to interpretation of retention parameters from HPLC systems comprising biomacromolecules. Role of lipophilicity in drug-biomacromolecule interactions is discussed in terms of quantitative structure-retention relationships (QSRR). Finally, reports are analyzed on systemic information which can be extracted by multivariate methods of data processing, like principal component analysis (PCA), from sets of lipophilicity parameters determined in diverse HPLC systems. PMID- 12570699 TI - Vascular actions of anthracycline antibiotics. AB - Anthracycline antibiotics are of particular value in the therapy of malignant diseases and exert profound effects not only on tumor cells but also on cells in the cardiovascular system. These quinone drugs affect vascular tone by a multitude of mechanisms, including acute modulation of Ca(2+) homeostasis, altered expression of membrane proteins and enzymes that are involved in the control of smooth muscle contraction, and generation of autoregulatory mediators, such as nitric oxide and endothelin. Anthracyclines interfere with blood coagulation-fibrinolysis balance due to its effects on the production of prostacyclin, plasminogen activator and plasminogen activator inhibitor in the endothelium. Moreover, anthracyclines are thought to be the modulators of angiogenesis. The intensity and quality of anthracycline actions on blood vessel function are highly variable and may depend not only on the chemical structure of anthracycline but also on the type of blood vessel as well as the metabolic and redox status of the vascular tissue. Vascular actions of anthracyclines are possibly involved in both beneficial as well as toxic and undesirable side effects such as tumor progress. Further investigations are required to clarify the relation between specific modifications of vascular cell function and clinical events observed during antineoplastic therapy with anthracyclines. PMID- 12570700 TI - Recent advances in peroxisome proliferator-activated receptor science. AB - The peroxisome proliferator-activated receptor (PPAR) family of nuclear receptors, a set of three receptor sub-types encoded by distinct genes, function as lipid sensors to regulate a broad range of genes in many metabolically active tissues. Synthetic PPAR agonists have exhibited therapeutic benefits in treating diabetes and cardiovascular diseases. The discovery of PPAR-specific ligands has led to significant advancement in our understanding of the structure of these receptor proteins and the molecular mechanism of their ligand-dependent activation. Herein, we present both recent progress in the functional analysis of these orphan receptors and the confirmation of the PPARs as molecular targets for the development of new medicines to treat human metabolic disease. PMID- 12570701 TI - Abuse of clenbuterol and its detection. AB - Clenbuterol and other beta-agonists are commonly misused as repartitioning agents in meat production and as doping substances to improve athletic performance. Numerous reports on food poisoning throughout Europe prompted the EU regulatory offices and FDA to implement a ban on the use of beta-agonists as growth promoters. Several analytical methods have been developed for detecting illegal administration of these compounds, based mainly on chromatography and immunoassay screening. This article deals with the pharmacological aspect of beta-agonists in growth promotion, control of their abuse and methods of analysis. PMID- 12570702 TI - Drugs that activate specific nitric oxide sensitive guanylyl cyclase isoforms independent of nitric oxide release. AB - Nitric oxide (NO) releasing drugs have helped patients suffering from angina pectoris for more than a century. In the 1970s NO-sensitive guanylyl cyclase was identified as the target of NO. Since then, three different isoforms of the enzyme have been identified. All NO-releasing drugs act by binding of NO to the prosthetic heme group common to all three isoforms. They thus act all as isoform unspecific substances. This review addresses recently developed drugs that activate NO-sensitive guanylyl cyclase independent of NO-release. They have great potential in the treatment of angina pectoris, hypertension and erectile dysfunction. The molecular target has been validated by the successful clinical use of NO-releasing drugs for more than a century. At the same time the mode of action of these drugs is entirely new. The development of highly isoform-specific derivatives with distinct pharmacological profiles is now an open possibility with great potential. PMID- 12570703 TI - Reservoirs of HIV replication after successful combined antiretroviral treatment. AB - Sustained reduction of viral replication can be achieved in HIV infected patients after treatment with combinations of drugs (HAART) that inhibit the viral reverse transcriptase, and protease enzymes. However, replication competent virus can still be recovered from latently infected resting memory CD4+ T-cell lymphocytes. Moreover, "covert" virus replication has been demonstrated in patients who experienced reductions in plasma viremia to levels below the limit of detection of the most sensitive PCR assays. In most studies, preferential attention has been given to latent resting CD4+ T-lymphocytes as a source of HIV persistence. However, insufficient suppression of HIV replication could also lead to viral re emergence after HAART interruption. In addition to CD4+ T- lymphocytes, other host cells such as long-lived resident macrophages or recently infected blood monocytes could also contribute to maintain persistent HIV replication after HAART. Establishing the origin of re-emerging HIV in patients under HAART upon treatment interruption is important to design optimal treatment schemes. Therapeutic strategies aimed at reducing the number of latently infected cells involve immune activation with IL-2, or other stimulatory factors, in the presence of antiretroviral drugs. Elimination of replication-competent virus would require intensification of HAART, or the use of antiretroviral drugs achieving an effective concentration at the site of HIV replication. In this review the mechanisms of HIV persistence and the methods that can be used to distinguish latent from covert HIV replication in different cell types will be discussed. PMID- 12570704 TI - Conformational analysis and molecular properties of N-(substituted phenyl carbonylamino)-4-(1-hydroxymethylphenyl)-1,2,3,6-tetrahydropyridines. AB - The three-dimensional structures of active derivatives of N (substitutedphenylcarbonylamino)-4-(1-hydroxymethylphenyl)-1,2,3,6-tetrahydropyri dines, which have previously been shown to possess anti-inflammatory activities, were built using BIOMEDCAche 5.0 software program. In addition, the three dimensional structures of some of the inactive ones were similarly generated. The conformational analysis, molecular and electronic structures were examined by molecular mechanics and quantum mechanics calculations. The primary objective was to clarify the effects of physicochemical properties of substituents on activity, since the exact role of the substitution pattern on the phenyl ring is uncertain. In addition, the experimental log P values did not appear to have any influence on the anti-inflammatory potencies of these compounds, since compounds having differing lipid solubilities are equiactive. We found that strongly electron donating group, such as the para-substituted methoxy group, detracts from activity. The conformational analysis indicated that the 4-ethyl derivative had the lowest energy conformation. Except for compound 1, which showed the lowest surface volume, compounds 2-9 had nearly similar surface volumes. PMID- 12570705 TI - Poly(ADP-ribose) polymerase inhibitors. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) is the principal member of the PARP enzyme family consisting of PARP-1 and several recently identified novel poly(ADP ribosyl)ating enzymes. PARP-1 functions as a DNA damage sensor and signalling molecule. Upon binding to DNA breaks, activated PARP cleaves NAD(+) into nicotinamide and ADP-ribose and polymerizes the latter onto nuclear acceptor proteins including histones, transcription factors and PARP itself. This Poly(ADP ribosyl)ation contributes to inflammatory signal transduction processes. In addition, oxidative stress-induced overactivation of PARP consumes NAD(+) and consequently ATP, culminating in cell dysfunction or necrosis. Activation of PARP has been implicated in the pathogenesis of stroke, myocardial ischemia, diabetes, diabetes-associated cardiovascular dysfunction, shock, traumatic central nervous system injury, arthritis, colitis, allergic encephalomyelitis and various other forms of inflammation. Therefore, inhibition of PARP by pharmacological agents may prove useful for the therapy of these diseases, as has been shown in preclinical animal models. Moreover, PARP inhibitors may have additional, potential utility as anticancer agents, radiosensitizers and antiviral agents. In the present article we overview the structures and pharmacological actions of various pharmacological classes of compounds which inhibit the catalytic activity of PARP. PMID- 12570706 TI - Advances in extracorporeal detoxification by MARS dialysis in patients with liver failure. AB - Although liver transplantation has become standard procedure for patients with liver failure, a number of issues in the management of these patients remains to be addressed. Alternative approaches have been tested, such as hepatocytes containing liver-support systems and filtration devices. However, the replacement of detoxification has been difficult, as the majority of toxins accumulating in liver failure is albumin-bound. Albumin dialysis (MARS system) is characterized by the specific removal of albumin-bound toxins through an innovative membrane transport. In particular, the albumin acts as a specific molecular adsorbent that is regenerated on line in a recycling system. Nowadays MARS represents the most frequently used liver support system. This treatment has been shown to remove albumin-bound molecules, such as bilirubin, bile acids, aromatic amino acids and copper. The removal of these toxins is clinically accompanied with an improvement of liver, cardiovascular and renal functions and hepatic encephalopathy. In several trials MARS was found to improve the clinical situation in patients with acute exacerbation of chronic liver failure and acute hepatic failure, but also in hepatorenal syndrome and primary graft non function or chronic rejection after liver transplantation. In summary, a critical analysis of the literature confirms that MARS device can be a safe therapeutic choice to achieve a better clinical outcome, and, sometimes, a survival advantage in patients with liver failure, even if a multi-center randomized trial is the only reliable way to enforce today's results. Further advances in the MARS components will definitively state whether albumin dialysis may represent the future in the field of artificial liver devices. PMID- 12570707 TI - Advances in the science of estrogen receptor modulation. AB - This work details recent advances in the science of estrogen receptor (ER) modulation, with emphasis on the discovery of novel ligands for the ER ligand binding domain (LBD). A detailed examination of structural studies of the ERs is presented with analysis of the impact of such works on contemporary ligand design and the molecular pharmacology of the ER. The various classes of ER modulators are discussed on the basis of stuctural similarities including selective estrogen receptor modulators (SERMs) and 'pure' non-steroidal antiestrogens. Additionally we review the emergence of a novel selective class of modulator which we have termed the selective estrogen receptor subtype modulators (SERSMs) and, in a departure from LBD strategies we examine the discovery of novel peptide inhibitors of the ER which inhibit transcriptional activiation of agonist liganded receptor through interaction with coactivator recruitment proteins, and offer unique insight to the mechanism of action of all classes of ER modulators. Through examination of patent and classical literature we present a thorough and informative cross-section of the contemporary state of the art in this exciting field of pharmaceutical research. PMID- 12570708 TI - Polyene macrolide antibiotics and their applications in human therapy. AB - Fungal infections represent a serious problem for patients with immune systems compromized either by HIV infection, or administration of immunosuppressive drugs during cancer therapy and organ transplantation. High dissemination and proliferation rates of many pathogenic fungi along with their insusceptibility to common antimicrobial drugs urge implementation of efficient and reliable antifungal therapy. Up to date, polyene macrolide antibiotics proved to be the most effective antifungal agents due to their potent fungicidal activity, broad spectrum, and relatively low frequency of resistance among the fungal pathogens. However, polyene macrolides are rather toxic, causing such serious side effects as renal failure, hypokalemia and thrombophlebitis, especially upon intravenous administration. Current views on the biosynthesis of polyene macrolides, their mode of action and structure-function relationship, as well as strategies used to overcome the toxicity problem are discussed in this review. In addition, some of the new potential applications for polyene macrolides in therapy of prion diseases, HIV infection and cancer are highlighted. PMID- 12570709 TI - Prediction of biological activity spectra for substances: evaluation on the diverse sets of drug-like structures. AB - The concept of Biological Activity Spectrum served as a basis for developing PASS (Prediction of Activity Spectra for Substances) software product. PASS predicts simultaneously more than 780 pharmacological effects and biochemical mechanisms based on the structural formula of a substance. It may be used for finding new targets (mechanisms) for known pharmaceuticals and for searching new biologically active substances. PASS prediction ability was evaluated by activity spectra prediction for 63 substances that are presented in the Molecule of the Month section of Prous Science (http://www.prous.com), belong to different chemical classes and reveal various types of biological activity. Mean accuracy of prediction turned out to be about 90%; therefore, it is reasonable to use PASS for finding and optimizing new lead compounds. A web-site with a new internet version of PASS is introduced into practice in December 2001 (http://www.ibmh.msk.su/PASS). On the site, one can find a detailed description of the PASS approach as well as some examples of its applications, and estimate the quality of prediction by submitting structures of substances with known activities. PMID- 12570710 TI - Receptor-operated regulation of ATP-sensitive K+ channels via membrane phospholipid metabolism. AB - ATP-sensitive K(+) channels (K(ATP)channels) regulate insulin secretion by coupling intracellular metabolic changes to excitability of the plasma membrane in pancreatic beta-cells. The channels are closed when extracellular glucose levels are elevated due to enhanced feature. By contrast, cardiac-type K(ATP) channels, which open in response to metabolic stress during cardiac ischemia, shorten action potential durations. This may contribute to the cardioprotection by decreasing Ca(2+) influx through sarcolemma. By sensing intracellular ATP levels or ATP/ADP ratios, changes in activity of K(ATP) channels convert metabolic information into membrane excitability. In addition to channel regulation by nucleotide concentrations, the channel activity is also dependent on the concentrations of membrane phospholipids, including phosphatidyl inositol 4,5-bisphosphate (PIP(2)). The levels of PIP(2) in the membrane may determine the basal activity of the channels. This suggests that channel activity would be modulated by the pathway of receptor-coupled GTP-binding protein (G-protein) and phosphatidyl inositol phospholipase C (PI-PLC) stimulation, which brings about depletion of the membrane PIP(2) pool. Thus, K(ATP) channels not only provide interface of metabolic changes with electrical excitation, but also rapidly transmit extracellular signals through receptor-coupled G-protein and PI-PLC pathway via PIP(2) metabolism. PMID- 12570711 TI - Gene silencing mediated by small interfering RNAs in mammalian cells. AB - RNA interference (RNAi) as a protecting mechanism against invasion by foreign genes was first described in C. elegans and has subsequently been demonstrated in diverse eukaryotes such as insects, plants, fungi and vertebrates. RNAi is the mechanism of sequence-specific, post-transcriptional gene silencing initiated by double-stranded RNAs (dsRNA) homologous to the gene being suppressed. dsRNAs are processed by Dicer, a cellular ribonuclease III, to generate duplexes of about 21 nt with 3'-overhangs (small interfering RNA, siRNA) which mediate sequence specific mRNA degradation. In mammalian cells siRNA molecules are capable of specifically silencing gene expression without induction of the unspecific interferon response pathway. Thus, siRNAs have become a new and powerful alternative to other genetic tools such as antisense oligonucleotides and ribozymes to analyze loss-of-function phenotypes. Application of siRNA duplexes to interfere with the expression of a specific gene requires knowledge of target accessibility, highly effective delivery of siRNAs into target cells and for some applications long-term siRNA expression. Effective strategies to deliver siRNAs to target cells in cell culture include transduction by physical or chemical transfection. An alternative strategy uses the endogenous expression of siRNAs by various Pol III promoter expression cassettes that allow transcription of functional siRNAs or their precursors. This review summarizes some genetic and biochemical aspects of RNAi, the delivery and application of siRNAs to target cells, the kinetics of RNAi and the utility of siRNAs as analytical and potential therapeutic tools. PMID- 12570712 TI - Cisplatin biochemical mechanism of action: from cytotoxicity to induction of cell death through interconnections between apoptotic and necrotic pathways. AB - Although cisplatin, cis-diamminedichloroplatinum(II), has been successfully used in the chemotherapy of cancer for more than 25 years, its biochemical mechanism of action is still unclear. The current accepted paradigm about cisplatin mechanism of action is that the drug induces its cytotoxic properties through binding to nuclear DNA and subsequent interference with normal transcription, and/or DNA replication mechanisms. If cisplatin-DNA adducts are not efficiently processed by cell machinery, cytotoxic processes eventually end up in cell death. However, before cisplatin enters the cell it may bind to phospholipids and phosphatidylserine in the cell membrane. In addition, in the cytoplasm many potential platinum-binding sites are also available, including RNA and sulfur containing biomolecules. Moreover, there is much evidence suggesting that the cytotoxic effects induced by binding of cisplatin to non-DNA targets (especially proteins) may contribute to its biochemical mechanism of action. On the other hand, it has been found that several factors such as the dose of drug as well as the metabolic condition of the cell subjected to cisplatin aggression, may determine that cancer cells die through apoptosis or necrosis. In fact, it has recently been reported that both mechanisms of cell demise work in concert so that within a population of tumour cells there is a continuum of possible modes of cell death. PMID- 12570713 TI - Prospects for pharmacologic inhibition of hepatic glucose production. AB - Type 2 diabetes is a widespread disease where effective pharmacologic therapies can have a profound beneficial public health impact. Increased hepatic glucose production (HGP) is observed in diabetics and its moderation by currently available agents provides therapeutic benefits. This review describes the challenges associated with the discovery of small molecules that inhibit HGP. Gluconeogenesis, glycogenolysis, liver architecture, and hepatocyte composition are described to provide background information on hepatic function. Current methods of target validation for drug discovery, HGP measurement, diabetes animal models, as well as current drug therapies are covered. In the accompanying review article the new drug targets being probed to produce the next generation of therapies are described. Significant pharmaceutical and academic efforts to pharmacologically inhibit HGP has the opportunity to provide new therapeutics for type 2 diabetics. PMID- 12570714 TI - Potential drug targets and progress towards pharmacologic inhibition of hepatic glucose production. AB - A number of therapeutic targets are currently under investigation for inhibition of hepatic glucose production with small molecules. Antagonists of the glucagon receptor, glycogen phosphorylase, 11-beta-hydroxysteroid dehydrogenase-1 and fructose 1,6-bisphosphatase are, or have been, under evaluation in human clinical trials. Other strategies, including glucocorticoid receptor antagonists and carnitine palmitoyltransferase inhibitors, are supported by proof of principle studies in man as well as rodents. Several potential targets including glucose-6 phosphatase, glucose-6-phosphatase translocase, glycogen synthase kinase-3, adenosine receptor 2B antagonists, phosphoenolpyruvate carboxykinase and pyruvate dehydrogenase kinase, have been validated by compounds that are effective in animal models. Other targets like PGC-1a and CREB have initial validation support but no medicinal chemistry has been reported. PMID- 12570715 TI - Chemical modification of glycyrrhizic acid as a route to new bioactive compounds for medicine. AB - Glycyrrhizic Acid (GL) is the major bioactive triterpene glycoside of licorice root (Glycyrrhiza Radix) extracts possessing a wide range of pharmacological properties (anti-inflammatory, anti-ulcer, anti-allergic, anti-dote, anti oxidant, anti-tumor, anti-viral etc.). Official sources of GL are Glycyrrhiza glabra L. and Gl. uralensis Fish. (Leguminosae). The content of GL in licorice root is 2-24% of the dry weight. GL is one of the leading natural compounds for clinical trials of chronic active viral hepatitis and HIV infections (preparation Stronger Neo-Minophagen C, SNMC), and its monoammonium salt (glycyram, tussilinar) is used as an anti-inflammatory and anti-allergic remedy. The synthetic transformations of GL on carboxyl and hydroxyl groups were carried out to produce new bioactive derivatives for medicine. GL esters were produced containing fragments of bioactive acids (4-nitrobenzoic, cinnamic, salycilic, acetylsalycilic, nicotinic, isonicotinic). Bioactive amides of GL were synthesized using chloroanhydride technique and N,N'-diciclohexylcarbodiimide (DCC) method. The synthesis of acylthioureids and semicarbazones was carried out via the reaction of triacylisothiocianate of penta-O-acetyl-GL with primary amines and hydrazines. The chain of transformations of trichloranhydride of penta O-acetyl-GL was made with the introduction of diazoketone groups in the molecule. A new group of GL derivatives to be triterpene glycopeptides was prepared by the activated esters method (N-hydrohysuccinimide-DCC or N-hydroxybenzotriazol-DCC) using alkyl (methyl, ethyl, propyl, butyl, tert-butyl) or benzyl (4-nitrobenzyl) esters of amino acids. The glycyrrhizyl analogs of the known immunostimulator, N acetyl-muramoyldipeptide (MDP), were synthesized using Reagent Woodward K. A series of ureids and carbamates of GL was synthesized containing 5-amino-5-desoxy D-xylopyranose units. The synthesis of 4-nitro-4-desoxy-glycosides, modified analogs of GL, was carried out by the oxidative splitting of the carbohydrate part of GL with NaIO(4). Triterpene 2-desoxy-D-glycosides, analogs of GL, were prepared by the glycal method in the presence of iodine-containing promoters or sulfonic acid cation-exchange resin KU-2-8 (H+) and LiBr. New anti-inflammatory and anti-ulcer agents were found among GL derivatives such as esters, amides, ureids, carbamates, thioureids and glycopeptides. GL glycopeptides are of interest as immunomodulators. Some of the chemically modified GL derivatives (salts, amides, glycopeptides) were potent HIV-1 and HIV-2 inhibitors in vitro. Preparation niglizin (penta-O-nicotinate of GL) was studied clinically as an anti inflammatory agent and is of interest for studies as hepatoprotector and HIV inhibitor. PMID- 12570716 TI - In vitro and in vivo antitumor effects of bisphosphonates. AB - Bisphosphonates are powerful inhibitors of osteoclast-mediated bone resorption. They are currently used in the palliative treatment of bone metastases. However, bisphosphonates do not only act on osteoclasts. There is now extensive in vitro preclinical evidence that bisphosphonates can act on tumor cells: they inhibit tumor cell adhesion to mineralized bone as well as tumor cell invasion and proliferation. Bisphosphonates induce also tumor cell apoptosis and stimulate gammadelta T cell cytotoxicity against tumor cells. In vivo, bisphosphonates inhibit bone metastasis formation and reduce skeletal tumor burden. This may reflect direct antitumor effects and indirect effects via inhibition of bone resorption. In addition, bisphosphonates inhibit experimental angiogenesis in vitro and in vivo. Understanding the molecular mechanisms through which bisphosphonates act on tumor and endothelial cells will be undoubtedly an important task in the future. It will allow the design of clinical trials to investigate whether the antitumor activity of bisphosphonates can be realized in the clinical setting. PMID- 12570717 TI - Natural and synthetic xanthonolignoids: chemistry and biological activities. AB - Natural xanthonolignoids are formed by the coupling of cinnamyl alcohol with an appropriate ortho-dihydroxyxanthone. More than twenty xanthonolignoids have been isolated and synthesized and some interesting pharmacological activities of these compounds have been described. The purpose of this review is to describe the natural occurrence, isolation, structure elucidation and synthesis - both biomimetic and classic - as well as biological activities of xanthonolignoids. Aspects related to biosynthesis will also be considered. PMID- 12570718 TI - Synergy in plant medicines. AB - The notion that medicines derived from plants depend for their action on active principles present has to be modified in view of the findings that there are, in many cases, adjuvant substances in the plant which enhance the activity of the components actually responsible for the effect. This synergy may involve protection of an active substance from degradation by enzymes; it may facilitate transport across barriers such as cell and organelle walls, it may overcome multi drug resistance mechanisms or provide other signals to the host's cells that result in higher efficacy of the crude drug when compared with isolated components. The many plant substances that stimulate the immune system, often at very low doses, have not been reviewed as this is not strictly speaking synergy. Some of the evidence for the phenomena described is reviewed and its bearing on phytotherapy commented. PMID- 12570719 TI - Biological activity and synthetic metodologies for the preparation of fluoroquinolones, a class of potent antibacterial agents. AB - In this present review we report different synthetic methodologies for the preparation of fluoroquinolones and their biological properties. The appearance of the fluoroquinolones, a new class of antibacterial agents (based on nalidixic acid, 4-quinolone-3-carboxylates), in early 1980's, gave a new impulse for the international competition to synthesize more effective drugs. Fluoroquinolones have a broad spectrum of activity against Gram-positive, Gram-negative and mycobacterial organisms as well as anaerobes. The fluoroquinolone ciprofloxacin hydrochloride is an important bioterrorist weapon and also an antibiotic used to treat bacterial infection in many different parts of the body, approved for use in patients who have been exposed to the inhaled form of anthrax. PMID- 12570720 TI - Lessons learned from the irinotecan metabolic pathway. AB - Irinotecan, a camptothecin analogue, is a prodrug which requires bioactivation to form the active metabolite SN-38. SN-38 acts as a DNA topoisomerase I poison. Irinotecan has been widely used in the treatment of metastatic colorectal cancer, small cell lung cancer and several other solid tumors. However, large inter patient variability in irinotecan and SN-38 disposition, as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan. Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy. Irinotecan is subjected to be shunted between CYP3A4 mediated oxidative metabolism to form two inactive metabolites APC or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by UGT1A1 to form SN-38G. The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan (e.g., CYP3A4, CYP3A5, UGT1A1) and sharing competitive elimination pathways with many concomitant medications, such as anticonvulsants, St. John's Wort, and ketoconazole. Efflux of the parent compound and metabolites out of cells by several drug transporters (e.g., Pgp, BCRP, MRP1, MRP2) also occurs. This review highlights the latest findings in drug activation, transport mechanisms, glucuronidation, and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent. PMID- 12570721 TI - "Multi-component reactions : emerging chemistry in drug discovery" 'from xylocain to crixivan'. AB - With the recent emergence of combinatorial chemistry and high-speed parallel synthesis for drug discovery applications, the multi-component reaction (MCR) has seen a resurgence of interest. Easily automated one-pot reactions, such as the Ugi and Passerini reactions, are powerful tools for producing diverse arrays of compounds, often in one step and high yield. Despite this synthetic potential, the Ugi reaction is limited by producing products that are flexible and peptide like, often being classified as 'non drug-like'. This review details developments of new, highly atom-economic MCR derived chemical methods, which enable the fast and efficient production of chemical libraries comprised of a variety of biologically relevant templates. Representative examples will also be given demonstrating the successful impact of MCR combinatorial methods at different stages of the lead discovery, lead optimization and pre-clinical process development arenas. This will include applications spanning biological tools, natural products and natural product-like diversity, traditional small molecule and 'biotech' therapeutics respectively. In particular, this review will focus on applications of isocyanide based MCR (IMCR) reactions. PMID- 12570722 TI - Controlled modulation of inflammatory, stress and apoptotic responses in macrophages. AB - An outstanding question of current immunology is to define the mechanisms by which microbial products influence the immunopathologic host response elements in the early stages of infection. Macrophages are now well recognized to have a critical role in both innate and acquired immunity. In order to adjust promptly to continuous changes in microenvironment and maintain the immunologic balance, macrophages adequately respond by activating one of the numerous immunologic programs. However, sustained macrophage activation and excessive production of inflammatory mediators can perpetuate the numerous pathological processes and contribute to induction of stress response and even apoptosis. Therefore, selective modulation of macrophage activity represents an important strategy for prevention and treatment of inappropriate inflammatory responses in order to minimize the unwanted side-effects of the immunity. Macrophages can be selectively reprogrammed for a specific phenotype of immune response, e.g. cytokine or nitric oxide (NO), by relatively short-term exposure of the cells to substimulatory concentrations of different microbial components, including LPS. These LPS-dependent reprogramming effects are mediated by IFN-gamma-independent autocrine cytokine regulatory mechanisms that also controlled at the transcriptional level. Furthermore, LPS reprogrammed macrophages exhibit differential capacity to resist experimentally induced apoptosis and to produce heat shock proteins. Complete analysis of, and appreciation for, the immunoregulatory mechanisms implicated in LPS-dependent reprogramming of immune responses in macrophages can be expected to increase our understanding of the host innate response, as well as allow investigators to utilize emerging immunologic technologies in effective treatment of infections and chronic inflammatory diseases. PMID- 12570723 TI - Unfolding the pathophysiological role of bioactive lysophospholipids. AB - Lysophospholipids (LPLs), including glycerol- and sphingoid-based lipids, stimulate cell signaling and play important pathophysiological roles in humans and other animals. These LPLs include lysophosphatidic acid (LPA), lysophosphatidylinositol (LPI), lysophosphatidylcholine (LPC), lysophosphatidylserine (LPS), sphingosine-1-phosphate (S1P), and sphingosylphosphorylcholine (SPC). Analyses of LPLs in human body fluids from subjects with different pathophysiological conditions reveal not only the relevance of LPLs in human diseases, but also their potential application as biomarkers and/or therapeutic targets. In recent years, the identification and/or characterization of the plasma membrane receptors for LPLs and enzymes regulating the metabolism of LPLs have greatly facilitated our understanding of their role and signaling properties. In vitro and in vivo functional and signaling studies have revealed the broad and potent biological effects of LPLs and the mechanisms of LPL actions in different cellular systems. Development of specific antagonists for each of the LPL receptors will provide powerful tools for dissecting signaling pathways mediated by receptor subtypes. More importantly, these antagonists may serve as therapeutics for relevant diseases. Genetic depletion of LPL receptors in mice has provided and will continue to provide critical information on the pathophysiological roles of LPL receptors. It is important to further evaluate the significance of targeting these bioactive LPL receptors, their downstream signaling molecules, and/or metabolic enzymes in the treatment of cancers and other diseases. PMID- 12570724 TI - Studies on human neutrophil biological functions by means of formyl-peptide receptor agonists and antagonists. AB - Phagocytes are activated by several extracellular signals, including formyl peptides derived from bacterial proteins or disrupted cells. The most intensely studied member of the formylpeptide family is the synthetic tripeptide N-formyl-L methionyl-L-leucyl-L-phenylalanine (fMLP), whose specific receptors have been identified on neutrophil plasma membrane and subsequently cloned. The fMLP receptor interaction activates multiple transduction pathways responsible for various neutrophil functions such as adhesion, chemotaxis, exocytosis of secretory granules and superoxide anion production, which represent the physiological response to bacterial infection and tissue damage. An unresolved question is whether signaling requirements are identical or specific for each physiological function. The development of fMLP receptor agonists and antagonists has led to an improvement of our knowledge about the above issue. Of particular interest is the possibility that receptorial antagonists, able to transiently inhibit neutrophil responses to formylpeptides, could be therapeutic agents in the treatment of inflammation-related diseases. Aim of this review is, i) to summarise the current understanding of the series of events that begins at the level of formylpeptide-receptor interaction and is responsible for the activation of transduction pathways, which finally determine neutrophil response; ii) to define the state of art regarding the synthesis as well as the biological actions of fMLP receptor agonists and antagonists. PMID- 12570725 TI - Vitamin D analogs--drug design based on proteins involved in vitamin D signal transduction. AB - Vitamin D analogs have proven to be very valuable tools for the treatment of calcium-related diseases and certain hyperproliferative conditions such as renal osteodystrophy, psoriasis and cancer. In general, vitamin D analogs exploit the enzymic and receptor machinery of the 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) signal transduction pathway. Key proteins in this cascade include the vitamin D receptor (VDR), the vitamin D-binding protein (DBP) and three cytochrome P450s (CYP27A, CYP27B and CYP24) which effect the synthesis and breakdown of the natural hormone, 1alpha,25(OH)(2)D(3). Analogs have been designed which reduce or enhance the importance of each of these proteins in the signal transduction pathway. Vitamin D prodrugs require one or more steps of activation and overcome congenital or acquired blocks in the 1alpha-hydroxylation step. By far the biggest class of vitamin D analogs are the VDR agonists which directly mimic 1alpha,25(OH)(2)D(3) and trigger protein conformational changes in the receptor which lead to changes in the transcriptional machinery at vitamin D responsive genes. Other emerging classes of molecules include the VDR antagonists and CYP24 inhibitors which target different events in the cascade. This review assesses the relative importance of each of the proteins of the vitamin D cascade, evaluates the success of these modifications in tailoring drugs in all classes for selected disease states and contemplates future directions for the field. PMID- 12570726 TI - WISH cells as a model for the "in vitro" study of amnion pathophysiology. AB - In the course of pregnancy amnion cells produce a number of factors which include cytokines and prostaglandins (PGs) produced in response to autocrine, paracrine and endocrine signals. Recent studies performed by several researchers contributed to elucidate the mechanism through which amnion tissue is involved in the triggering of physiological labor. However, there are other possible functions to be ascribed to amniotic cells, depending on the high number of factors that they produce as well as on the receptors that enable them to act in turn as target. For instance, it has been demonstrated that amnion cells are able to produce lecithin upon the regulation of several factors, such as glucocorticoids and epidermal growth factor, a finding that suggests a protective role of the tissue on fetal pulmonary function. As regards to triggering the uterine contractions, it is accepted that prostaglandin release by amnion cells represents a key event. It is under the control of hormones, growth factors, cytokines and probably PGs themselves. A striking analogy has been found between the mechanism of inflammation and the onset of myometrial activity in labor. In this context, it has been shown that for-Met-Leu-Phe (fMLP), the prototype of a series of formylated peptides traditionally considered chemotactic agents, is also involved in the regulation of amniotic PG release. The similitude between labor and inflammatory response is enforced by the antiprostaglandin action of some classes of antibiotics observed in amnion tissue, that enable them as effective tools against preterm labor, both in the absence and in the presence of infection. As for the mechanisms responsible for the regulation of PG synthesis, some agents act by influencing protein synthesis, while others exert their effects through the production of intracellular second messengers, mainly represented by phosphatidyl-inositol-4-5 bisphosphate and cyclic AMP. The mechanism whereby second messengers induce PG release is not clear, and a crosstalk between the two transduction pathways could be hypothesized. This interaction has extensively been analysed in "WISH" cells, a human amnion-derived cell line, which represent a model for the in vitro study of amnion functions. In the present review, we intend to report the results of the studies regarding the mechanisms through which the control of the above mentioned functions is executed. PMID- 12570727 TI - Pathobiology of cholesterol gallstone disease: from equilibrium ternary phase diagram to agents preventing cholesterol crystallization and stone formation. AB - The primum movens in cholesterol gallstone formation is hypersecretion of hepatic cholesterol, chronic surpersaturation of bile with cholesterol and rapid precipitation of cholesterol crystals in the gallbladder from cholesterol enriched vesicles. Associated events include biochemical defects (increased biliary mucin, and increased proportions of hydrophobic bile salts in the intestine and gallbladder), motility defects (gallbladder smooth muscle hypocontractility in vitro and gallbladder stasis in vivo, sluggish intestinal transit), and an abnormal genetic background. The study of physical-chemical factors and pathways leading to cholesterol crystallization in bile has clinical relevance and the task can be carried out in different ways. The lithogenicity of bile is investigated in artificial model biles made by three biliary lipids - cholesterol, bile salts and phospholipids - variably combined in systems plotting within the equilibrium ternary phase diagram; also, crystallization propensity of ex vivo incubated human bile is studied by biochemical analysis of precipitated crystals, polarizing quantitative light microscopy and turbidimetric methods. The present review will focus on the recent advances in the field of pathobiology of cholesterol gallstones, by underscoring the role of early events like water transport, lipid transport, crystallization phenomena - including a genetic background - in gallstone pathogenesis. Agents delaying or preventing precipitation of cholesterol crystals and gallstone formation in bile will also be discussed. PMID- 12570728 TI - Mode of action of plant defensins suggests therapeutic potential. AB - Higher vertebrates can rely both on an innate as well as an adaptive immune system for defense against invading pathogens. In contrast, plants can only employ an innate immune system that largely depends on the production of antimicrobial compounds such as plant defensins and other pathogenesis-related proteins. Plant defensins are ubiquitous, cationic, cysteine-rich plant peptides and have a folding pattern that shares high similarity to defense peptides of mammals and insects, suggesting an ancient and conserved origin. A large number of plant defensins appear to display antifungal activity. Some of these defensins have been found to interact with fungal-specific components in the plasmamembrane, resulting in membrane permeabilization. This makes them an attractive source of potential therapeutics to treat fungal infections. PMID- 12570729 TI - Beta-lactamases: a survey of protein diversity. AB - Bacterial resistance to beta-lactam antibiotics and beta-lactamase inhibitors is an ever increasing problem that threatens the clinical utility of drugs that form the cornerstone of the antibiotic armamentarium. Especially among Gram-negative pathogens, elaboration of structurally and mechanistically novel beta-lactamase enzymes is the most important means by which resistance occurs. An appreciation of the tremendous diversity of these drug-modifying enzymes will assist in understanding why so few generally effective inhibitory agents exist for these unique drug targets. This review will give a general background on the reaction mechanisms and classification schemes of the more than 340 beta-lactamase enzymes described to date. A discussion will follow highlighting the emerging Class A SHV and TEM-derived extended-spectrum (ESBLs), and inhibitor-resistant enzymes, non TEM, non-SHV Class A ESBLs, and carbapenemases, Class B metallo-beta-lactamases and some of their novel inhibitors, plasmid and chromosomally encoded Class C enzymes, and finally, the OXA-type oxacillinases, ESBLs, and carbapenemases of Class D. The clinical importance of multiple resistance mechanisms in conjunction with the production of beta-lactamase enzymes is emphasized. PMID- 12570730 TI - Therapy of chronic hepatitis C: a critical review. AB - Combination therapy (Interferon plus ribavirin) is the current therapeutic gold standard for naive Hepatitis C Virus (HCV)-positive patients and with the recent advent of pegylated (PEG) IFN the rate of the sustained virological response (HCV RNA clearance 6 months after the end of treatment) is about 54%-56% with a therapeutical gain mainly among patients with unfavourable HCV genotype (1a, 1b); in this subset of patients, a 42%-46% sustained response rate is achieved compared with 33%-36% found among genotype 1 patients treated with the standard therapy. Patients who respond to IFN monotherapy but relapse during the follow-up should be re-treated with combination therapy given for at least 6 months at the minimum dose of 3 MU thrice weekly plus ribavirin 1000 mg/daily. Recent data suggest that prolonging the time of treatment (12 months) may induce a significantly higher rate of sustained response among patients with genotype 1. The efficacy of the combination of IFN and ribavirin in retreating patients with chronic hepatitis C not responding to IFN monotherapy is controversial as it ranges between 0% and 40%. Recent data show an overall rate of sustained response of 23% when an aggressive approach is adopted but increasing the dosage and the time of treatment induces a significant therapeutic benefit only in patients with genotype 1. In conclusion, a therapeutic progress for chronic hepatitis C has been achieved during the last 10 years (56% vs 20% of sustained response rate obtained with IFN monotherapy) but several unresolved issues are yet to be addressed. PMID- 12570731 TI - Strategy of computer-aided drug design. AB - Modern strategies of computer-aided drug design (CADD) are reviewed. The task of CADD in the pipeline of drug discovery is accelerating of finding the new lead compounds and their structure optimization for the following pharmacological tests. The main directions in CADD are based on the availability of the experimentally determined three-dimensional structure of the target macromolecule. If spatial structure is known the methods of structure-based drug design are used. In the opposite case the indirect methods of CADD based on the structures of known ligands (ligand-based drug design) are used. The interrelationship between the main directions of CADD is reviewed. The main CADD approaches of molecule de novo design and database mining are described. They include methods of molecular docking, de novo design, design of pharmacophore and quantity structure-activity relationship models. New ways and perspectives of CADD are discussed. PMID- 12570732 TI - Antivirals at the mirror: the lack of stereospecificity of some viral and human enzymes offers novel opportunities in antiviral drug development. AB - The enantioselectivity of enzymes, namely the property of enzymes to recognise and metabolise only one of the two enantiomers of chiral molecules, is related to the chiral structure of the enzymes, reflecting the three-dimensional folding of the polypeptide backbone and the orientation of the amino acid side chains in the folded molecule. Because of the chirality of the amino acids (L), the chemistry of life should be highly sensitive to different enantiomers of chiral substrates. However, in a world consisting only of D-nucleosides and L-amino acids, an enzyme which lacks enantio-selectivity does not reduce its fitness, since there is no chance of molecular misunderstanding when no other choice is available. Thus, although enantioselectivity is theoretically essential for life we do not expect to be always present among the biochemical properties of enzymes. If this is the case for key enzymes involved in virus infection or cancer, how to exploit such lack of enantioselectivity for a novel approach to antiviral or anticancer chemotherapy? The present review will discuss the possible lack of enantioselectivity of enzymes and its relevance for the developing of novel drugs with the inverted optical configuration. PMID- 12570733 TI - A review of macrolide treatment of atherosclerosis and abdominal aortic aneurysms. AB - Seroepidemiological studies have shown an association between Chlamydia pneumoniae and atherosclerosis, the risk of acute myocardial infarction and abdominal aortic aneurysms (AAA). Several studies have detected C. pneumoniae in atherosclerotic lesions from coronary and carotid arteries, in AAA, and in sclerotic aortic valves. However, culturing of C. pneumoniae is difficult and has seldomly succeeded from atherosclerotic lesions. Thus, the pathogenicity is unknown, and the significance of detecting the organism is unresolved. Nevertheless, in a large observational study comparing the risk of cardiovascular events among recipients of macrolide versus pencillins, macrolide treatment reduced the risk of such events after relevant adjustment. Furthermore, in two out of three minor randomized clinical trials were patients with ischaemic heart disease were randomized into antibiotic treated and placebo groups, a significant reduction in serious end-points were noticed in patients receiving macrolide. Similarly, two other minor randomized trials showed that macrolide treatment inhibited growth of small AAA. Macrolide therapy thus seems potential to improve the outcome of severe ischaemic heart disease, and growth of AAA. If true, it not known whether this is transient because of macrolide's non-specific anti inflammatory effect or latent infection, or permanent because of eradicating C. pneumoniae organisms. In order to clarify this, large and long term randomized trials are needed, as well as diagnostic methods that can differentiate between individuals who are or are not infected with C. pneumoniae. The latter are needed in order to clarify the impact of the presence of C. pneumoniae and to avoid overconsumption of antimicrobials, which can result in serious ecological problems. PMID- 12570734 TI - New approaches for anti-infective drug discovery: antibiotics, vaccines and beyond. AB - Infectious disease is the leading cause of death worldwide, and billions of dollars are invested every year in developing anti-infective drugs. In the meantime, resistant bacteria are on the steady rise and render many once effective drugs useless. The tremendous funding and the urgent need to treat the resistant bacterial infections lead to the rapid progress on development of new drugs and potential new drug targets. New discoveries are being made that increase our understanding of microbial pathogenesis. Technological advancement is also being made to accelerate the drug discovery process. This review will mainly focus on discussing novel strategies on the development of antibiotics and vaccines for treating bacterial infections. Details of how some of the emerging technologies such as genomics and bioinformatics are accelerating the drug discovery process will be highlighted. Newly emerging concepts in controlling bacterial infections such as the use of probiotics and enzybiotics will also be briefly described. PMID- 12570735 TI - Chitin synthesis as target for antifungal drugs. AB - Human mycoses have become a threat to health world-wide. Unfortunately there are only a limited number of antimycotic drugs in use. Promising targets for drugs specific against fungi are those affecting chitin synthesis. Chitin is absent in vertebrates, and is essential for fungal wall integrity. A thorough knowledge of the mechanism of chitin synthesis is required to design specific inhibitors. We review here our current understanding of the process, and the most promising drugs that inhibit it. Chitin is made by chitin synthases requiring specific microvesicles, the chitosomes, for intracellular transport. Fungi contain several chitin synthases, some of which may be essential at a certain stage. This phenomenon is important to take into account for drug design. The most widely studied chitin synthase inhibitors are polyoxins and nikkomycins that probably bind to the catalytic site of chitin synthases. These are not equally susceptible to the drugs. In Saccharomyces cerevisiae the order of sensitivity is: Chs3p>Chs1p>Chs2p. Main problems for their succesful use in vivo are: low permeability, and different susceptibility of fungal species, and variable responses in animal models. Chemical modifications have been proposed to make more potent derivatives. Other synthetic or natural compounds are also promising as possible inhibitors, but their properties are less well known. Rational drug design has proceeded only on the basis of existing inhibitors, because the structure of the active site of chitin synthase is unknown. Undoubtedly, determination of this, and the biosynthetic mechanism will reveal unexpected drug targets in the future. PMID- 12570736 TI - Bioinformatics and the discovery of novel anti-microbial targets. AB - Genomic research is playing a critical role in the discovery of new anti microbial drugs. The rapid increase in bacterial and eukaryotic genome sequences allows for new and innovative ways for obtaining antimicrobial protein targets. Here, we describe a two level strategy for target identification and validation using computers (in silico). First, large scale comparative analyses of genome sequences were used to identify highly conserved genes which might be essential for in vitro and/or in vivo survival of bacterial pathogens. Lab-based experiments provided confirmation or validation of the hypothesis of in silico essentiality for over 350 individual genes. Over 200 validated, broad spectrum; yet highly specific gene targets, were identified in community infection pathogens. The second part of the target discovery strategy is an in-depth evolutionary, structural and cellular analysis of key drug targets. As an example, phylogenetic and structural analyses suggest that sequence and binding pocket conservation in FabH (beta-ketoacyl-ACP synthase III) would allow for the development of small molecule inhibitors not only effective against a broad species spectrum of community bacterial pathogens but also as potential new therapies for tuberculosis and malaria. PMID- 12570737 TI - Novel antibacterials: a genomics approach to drug discovery. AB - The appearance of antibiotic resistant pathogens, including vancomycin resistant Staphylococcus aureus, in the clinic has necessitated the development of new antibiotics. The golden age of antibiotic discovery, in which potent selective compounds were readily extracted from natural product extracts is over and novel approaches need to be implemented to cover the therapeutic shortfall. The generation of huge quantities of bacterial sequence data has allowed the identification of all the possible targets for therapeutic intervention and allowed the development of screens to identify inhibitors. Here, we described a number of target classes in which genomics has contributed to its identification. As a result of analyzing sequence data, all of the tRNA synthetases and all of the two-component signal transduction systems were readily isolated; which would not have been easily identified if whole genome sequences were not available. Fatty acid biosynthesis is a known antibacterial target, but genomics showed which genes in that pathway had the appropriate spectrum to be considered as therapeutic targets. Genes of unknown function may seem untractable targets, but if those that are broad spectrum and essential are identified, it becomes valuable to invest time and effort to determine their cellular role. In addition, we discuss the role of genomics in developing technologies that assist in the discovery of new antibiotics including microarray gridding technology. Genomics can also increase the chemical diversity against which the novel targets can be screened. PMID- 12570738 TI - Genomic pathways to antifungal discovery. AB - The limitations of the therapeutic antifungals are becoming increasingly apparent in the clinic due to their modest efficacy against life-threatening systemic fungal infections. These antifungals belong to only a few structural classes that affect a small range of targets, some are quite toxic in humans while the use of others, particularly the azole drugs, has encouraged the emergence of resistant clinical isolates and the selection of innately resistant fungal pathogens. Only a few new drugs based on novel targets are in clinical development, and these may be insufficient to overcome the changing tide of fungal disease. In parallel with the successful completion of the Saccharomyces cerevisiae and human genome sequencing projects, an increasing number of genome sequencing projects are being initiated and completed for significant fungal pathogens. The growing repository of genomic information, which is complemented by decades of genetic and biochemical study, is now available for genome-wide analysis of gene function and for incisive inter-genomic comparison, with the S. cerevisiae and human genomes providing key points of reference. Functional genomic and comparative genomic techniques, many of which were developed with S. cerevisiae, are being applied to fungal pathogens with the aim of obtaining an integrated view of fungal biology and to extract targets suitable for drug discovery. This review describes some of these techniques, their limitations and their increasing contribution to the antifungal discovery process through effective gene annotation, target identification and prioritization, and in the optimization of antifungal leads. PMID- 12570739 TI - Novel intervention strategies for Helicobacter pylori treatment. AB - Helicobacter pylori infects the gastric mucosa of almost half of the worlds population and infection is associated with several gastrointestinal diseases, ranging in severity from superficial and chronic gastritis to duodenal ulceration and gastric adenocarcinoma. Developing new therapeutics against a bacterium with such a unique niche has proven challenging, and the current therapy is complex and increase of bacterial resistance to current antimicrobials and treatment failure has identified a need for newer, more potent compounds. Access to the genomic sequence of several H. pylori isolates has allowed a more focused, target specific approach to the development of new therapeutics. PMID- 12570740 TI - Assisting functional assignment for hypothetical Heamophilus influenzae gene products through structural genomics. AB - The three-dimensional structures of Haemophilus influenzae proteins whose biological functions are unknown are being determined as part of a structural genomics project to ask whether structural information can assist in assigning the functions of proteins. The structures of the hypothetical proteins are being used to guide further studies and narrow the field of such studies for ultimately determining protein function. An outline of the structural genomics methodological approach is provided along with summaries of a number of completed and in progress crystallographic and NMR structure determinations. With more than twenty-five structures determined at this point and with many more in various stages of completion, the results are encouraging in that some level of functional understanding can be deduced from experimentally solved structures. In addition to aiding in functional assignment, this effort is identifying a number of possible new targets for drug development. PMID- 12570741 TI - Natural and acquired macrolide resistance in mycobacteria. AB - The genus Mycobacterium contains two of the most important human pathogens, Mycobacterium tuberculosis and Mycobacterium leprae, the etiologic agents of tuberculosis and leprosy, respectively. Other mycobacteria are mostly saprophytic organisms, living in soil and water, but some of them can cause opportunistic infections. The increasing incidence of tuberculosis as well as infections with non-tuberculous mycobacteria (NTM) in AIDS patients has renewed interest in molecular mechanisms of drug resistance in these pathogens. Mycobacteria show a high degree of intrinsic resistance to most common antibiotics. For instance, species from the M. tuberculosis complex (MTC) are intrinsically resistant to macrolides. Nevertheless, some semi-synthetic macrolides as the erythromycin derivatives clarithromycin, azithromycin and most recently the ketolides, are active against NTM, particularly Mycobacterium avium, and some of them are widely used for infection treatment. However, shortly after the introduction of these new drugs, resistant strains appeared due to mutations in the macrolide target, the ribosome. The mycobacterial cell wall with its specific composition and structure is considered to be a major factor in promoting the natural resistance of mycobacteria to various antibiotics. However, to explain the difference in macrolide sensitivity between the MTC and NTM, the synergistic contribution of a specific resistance mechanism might be required, in addition to possible differences in cell wall permeability. This mini-review summarizes the current knowledge on the natural and acquired macrolide resistance in mycobacteria, gives an overview of potential mechanisms implicated in the intrinsic resistance and brings recent data concerning a macrolide resistance determinant in the MTC. PMID- 12570742 TI - Retinoic acid metabolism and mechanism of action: a review. AB - Retinoids are vitamin A (retinol) derivatives essential for normal embryo development and epithelial differentiation. These compounds are also involved in chemoprevention and differentiation therapy of some cancers, with particularly impressive results in the management of acute promyelocytic leukemia (APL). Although highly effective in APL therapy, resistance to retinoic acid (RA) develops rapidly. The causes of this resistance are not completely understood and the following factors have been involved: increased metabolism, increased expression of RA binding proteins, P-glycoprotein expression, and mutations in the ligand binding domain of RARalpha. RA exerts its molecular actions mainly through RAR and RXR nuclear receptors. In addition to the nuclear receptor based mechanism of RA action, covalent binding of RA to cell macromolecules has been reported. RA derives from retinol by oxidation through retinol and retinal dehydrogenases, and several cytochrome p450s (CYPs). RA is thereafter oxidized to several metabolites by a panel of CYPs that differs for the different RA isomers. Phase II metabolism, mainly glucuronidation, is also observed. The role RA metabolism plays in the expression of its biological actions is not completely understood: in several systems, metabolism decreases RA activity, whereas in other systems metabolism appears involved in its action. In addition, several RA metabolites have shown activity and cannot be classified as only catabolites. Therapy of cancer with retinoids is still in its infancy, but the use of new analogues with improved pharmacological properties, along with combination with other drugs, could undoubtedly improve the management of several cancers in the future. PMID- 12570743 TI - DNA demethylating agents and chromatin-remodelling drugs: which, how and why? AB - DNA hypermethylation at the CpG dinucleotides clustered in "islands" in the promoter regions of genes causes transcriptional repression through the remodelling of chromatin. Aberrant methylation patterns of tumor suppressor genes and their subsequent silencing constitute a common feature of many cancers. Thus, the search for drugs that interfere in methylation-mediated gene repression has become one of the major goals in the design of cancer therapies. The major actors in the mammalian methylation system are DNA-methyltransferases (DNMTs), and methyl-CpG-binding proteins (MBDs), which recognize methylated cytosine and recruit repressor complexes, including histone deacetylases (HDACs). In this context, two major groups of drugs can be distinguished. The first one is constituted by substances that inhibit the action of DNMTs, either competing with cytosine or with S-adenosylmethionine (SAM, AdoMet) or acting over the DNMTs themselves. The second group involves compounds that inhibit subunits of the repressor complexes, such as HDACs. In this manuscript we review these two different groups of drugs, discussing their properties and the side effects that have been described (that occur by interference with other metabolic pathways). We also propose the logical pharmacological extension of these findings to design more specific and effective drugs for the prevention and treatment of human cancer. PMID- 12570744 TI - Drug metabolism and individualized medicine. AB - Drug metabolism refers to the biochemical transformation of a compound into another more polar chemical form. Absorption, distribution, metabolism and excretion comprise an integral part in understanding the safety and efficacy of a potential new drug. Detailed in-depth knowledge of the Pharmacokinetics and Drug Metabolism of a new drug entity is considered a prerequisite to know the appropriate route of administration, correct dose etc. Sometimes there is (are) different/unwanted effect(s) of certain drugs in different populations. This is particularly true for the drug having narrow therapeutic index. Often these different effects are detrimental to an individual, thus termed as adverse drug reactions. After the raw draft of human genome has evolved, it has become increasingly clear that change(s) in the drug response between individuals, is due to the occurrence of genetic polymorphisms in the Phase I and II drug metabolizing enzymes, due to which distinct subgroups in the population differ in their ability to perform certain drug biotransformation reactions. The study about the occurrence of genetic polymorphisms in drug metabolizing enzymes is termed as Pharmacogenetics/ Pharmacogenomics. Pharmacogenetic characterization of particular drug can be both phenotypically or genotypically conducted in population groups. The study is very important to check the post-marketed drug withdrawal, if a particular percentage of population suffers from adverse drug reactions, and thus a lot of revenue be saved. The study also helps to find out Right Medicine for Right Individual or Individualized Medicine. PMID- 12570745 TI - Cancer and phase II drug-metabolizing enzymes. AB - Cancer development results from the interaction between genetic factors, the environment, and dietary factors have been identified as modulators of carcinogenesis process. The formation of DNA adducts is recognized as the initial step in chemical carcinogenesis. Accordingly, blocking DNA adducts formation would be the first line of defense against cancer caused by carcinogens. Glutathione-S-transferases inactivate chemical carcinogens into less toxic or inactive metabolite through reduction of DNA adducts formation. There are many different types of glutathione S-transferase isozymes. For example, GST delta serves as a marker for hepatotoxicity in rodent system, and also plays an important role in carcinogen detoxification. Therefore, inhibition of GST activity might potentiate the deleterious effects of many environmental toxicants and carcinogens. In addition, approximately half of the population lacks GST Mu expression. Epidemiological evidence showed that persons possessing this genotype are predisposed to a number of cancers including breast, prostate, liver and colon cancers. In addition, individual risk of cancer depends on the frequency of mutational events in target oncogenes and tumor suppressor genes which could lead to loss of chromosomal materials and tumor progression. The most frequent genetic alteration in a variety of human malignant tumors is the mutation of the coding sequence of the p53 tumor suppressor gene. O(6)-alkylguanine in DNA leads to very high rates of G:C deltaA:T transitions in p53 gene. These alterations will modulate the expression of p53 gene and consequently change DNA repair, cell division, and cell death by apoptosis. Also, changes in the expression of BcI-2 gene results in extended viability of cells by over-riding programmed cell death (apoptosis) induced under various conditions. The prolonged life-span increases the risk of acquiring genetic changes resulting in malignant transformation. In addition, a huge variety of food ingredients have been shown to affect cell proliferation rates. They, therefore, may either reduce or increase the risk of cancer development and progression. For example, it has been found that a high intake of dietary fat accelerates the development of breast cancer in animal models. Certain diets have been suggested to act as tumor promoters also in other types of cancer such as colon cancer, where high intake of fat and phosphate have been linked to colonic hyper-proliferation and colon cancer development. Different factors such as oncogenes, aromatic amines, alkylating agents, and diet have a significant role in cancer induction. Determination of glutathione S transferase isozymes in plasma or serum could be used as a biomarker for cancer in different organs and could give an early detection. PMID- 12570746 TI - A nuclear receptor-mediated xenobiotic response and its implication in drug metabolism and host protection. AB - Regulation of the Phase I CYP enzymes and Phase II conjugating enzymes is implicated in both drug metabolism and drug-drug interactions. Moreover, the elimination of numerous xenobiotic and endobiotic toxic chemicals also requires a concerted function of Phase I and II enzymes, as well as the membrane spanning drug transporters. The genes that encode these enzymes and transporters are inducible by numerous xenobiotics, yet the inducibility shows clear species specificity. In the last 3-4 years, orphan nuclear receptors (NRs) such as PXR, CAR, and FXR have been established as species-specific xeno-sensors that regulate the expression of Phase I and II enzymes, as well as selected drug transporters. This transcriptional regulation is achieved by binding of these xenobiotic receptors to the NR response elements found within the promoter regions of target genes. The identification of NRs as xenosensors represents a major step forward in understanding the genetic mechanisms controlling the expression of drug metabolizing enzymes. The establishment of NR-mediated and mechanism-guided xenobiotic screening systems by using cultured cells or genetically engineered mouse models has not only advanced our understanding of the molecular complexity of this drug-induced xenobiotic response, but has also provided in vitro and in vivo platforms to facilitate the development of safer drugs. PMID- 12570747 TI - Kidney CYP450 enzymes: biological actions beyond drug metabolism. AB - Arachidonic acid can be metabolized by cytochrome p450 (CYP450) enzymes to 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acids (EETs), their corresponding dihydroxyeicosa-trienoic acids (DHETs), and 20-hydroxyeicosatetraenoic acid (20 HETE). These arachidonic acid metabolites are involved in the regulation of renal epithelial transport and vascular function. 20-HETE and EETs are produced in the renal microvascular smooth muscle cells and endothelial cells, respectively. 20 HETE constricts the preglomerular arterioles by inhibiting K(+) channels, and contributes importantly to renal blood flow autoregulatory responsiveness of the afferent arterioles. EETs dilate the preglomerular arterioles by activating the renal smooth muscle cell Ca(2+)-activated K(+) channels and hyperpolarizing smooth muscle cells. These EET actions are consistent with their identification as endothelium-derived hyperpolarizing factors (EDHFs). In the kidney, EETs and 20-HETE are also produced in the proximal tubule and the thick ascending loop of Henle, and these metabolites modulate ion transport in the proximal tubules and the thick ascending limb by inhibiting Na(+)-K(+)-ATPase and the Na(+)-K(+)-2Cl( ) cotransporter. CYP450 metabolites also act as second messengers for many paracrine and hormonal agents, including endothelin, nitric oxide, and angiotensin II. The production of kidney CYP450 arachidonic acid metabolites is altered in diabetes, pregnancy, hepatorenal syndrome, and in various models of hypertension, and it is likely that changes in this system contribute to the abnormalities in renal function that are associated with many of these conditions. PMID- 12570748 TI - Developing the molecular modelling of diffusion in zeolites as a high throughput catalyst screening technique. AB - Molecular modelling techniques have been used to screen zeolite catalysts for their suitability for organic synthesis. For example, we have used these techniques to study the alkylation of aromatic molecules which are important in the fine-chemical and drug industries. A survey of all such efforts is reviewed in this article. The application of molecular modelling techniques in a systematic manner is an efficient first step in the design of zeolite catalysts. As a qualitative screening tool, molecular graphics is used to visualize how well the reactant and product molecules fit inside the pores of the zeolites. Using a hybrid of several molecular modelling methods, which combines molecular dynamics (MD) and Monte Carlo methods with energy minimization, it is possible to determine the minimum energy locations of the molecules inside the zeolites cages. The minimum energy configurations determined by this hybrid method are taken as a starting point for diffusion of the molecules through the zeolite channels. When a molecule is allowed to diffuse through zeolite channel, the molecule attains some maxima and minima in its diffusion energy profile. From the differences between a maximum and a minimum energy configuration, the diffusion energy barrier for the molecule can be calculated in the zeolites. By comparing the diffusion energy barriers for various isomers of a molecule in different zeolites, it is possible to find out the most suitable zeolite for achieving the required shape-selectivity. In addition, factors influencing the diffusivity of the molecules and consequently the shape selectivity are derived. The list of factors and their relative importance are analysed to derive valuable guidelines to design shape-selective zeolite catalysts for a given reaction. Thus, the ultimate aim of these studies is to develop a high throughput computational screening process for the selection of shape-selective zeolite catalysts for various reactions. The dynamic behaviour of molecules inside the pores of zeolites can be studied using MD methods. Since MD is computationally time consuming, it is more efficient to screen the possible zeolite catalysts by energy minimization methods and then perform MD in specific zeolites. More accurate values of diffusivity of the molecules can be calculated using MD methods, and these values can be correlated with the shape-selectivity observed experimentally and /or derived from diffusion energy barrier calculations. PMID- 12570749 TI - Purification strategies for combinatorial and parallel chemistry. AB - This review surveys the methods developed for the purification of intermediates and final compounds originating from parallel and combinatorial chemistry. Included will be reviews of polymer-assisted purification, liquid-phase combinatorial chemistry, fluorous synthesis, liquid-liquid and solid-phase extraction, reverse-phase HPLC and supercritical fluid chromatography. A critique of each method is given, highlighting the methodologies strengths and weaknesses. PMID- 12570750 TI - Use of intein-directed peptide biosynthesis to improve serum stability and bioactivity of a gelatinase inhibitory peptide. AB - Screening of phage display libraries allows rapid identification of peptides binding to a target. However, functional analysis of the phage sequences and their reproduction as soluble and stable peptides are often the most time consuming part in the screening. We have used here intein-based peptide biosynthesis to produce a phage-display derived gelatinase inhibitory peptide CTTHWGFTLC and to identify the critical residues for gelatinase inhibitory activity by performing alanine-scanning mutagenesis. By biosynthetic incorporation of 5-fluorotryptophan, we obtained an inhibitor of MMP-2 and MMP-9 gelatinases that showed a 6-fold enhancement in serum stability in comparison to the wild-type peptide. The new peptide also had an improved ability to inhibit tumor cell migration. These studies indicate the utility of intein methodology for synthesis and design of peptides obtained by phage display. PMID- 12570751 TI - DBU-assisted cyclorelease elimination: combinatorial synthesis and gamma-glutamyl cysteine synthetase and glutathione-S-transeferase modulatory effect of C nucleoside analogs. AB - A combinatorial library of 60C- nucleoside analogs was synthesized by sequential coupling of building blocks followed by cyclative cleavage with DBU in an efficient manner. Only DMSO soluble compounds were tested for their modulatory effect against filarial gamma-glutamyl cysteine synthetase (gamma-GCase) and glutathione-S-transeferases (GSTs). Several compounds were found to be weak inhibitors of filarial gamma-GCase, whereas, most of them stimulated filarial GSTs. PMID- 12570753 TI - Design and peptide-based validation of phage display antibodies for proteomic biochips. AB - To validate potential application of phage display-antibody arrays for high throughput screening on a novel proteomics biochip, we examined the epitopes versus the full protein of glucose-6-phosphate-dehydrogenase (G6PD) from yeast. In a predictive approach, we used the Hopp-Woods method and compared the results with antibodies directed against the entire enzyme. In total, 16 peptides of a length of 11 amino acids each fulfilling the desired criteria were identified and synthesized. Subsequently, antibodies against G6PD were raised using a phage display library. Selective interaction of the antibodies with certain peptides facilitated the identification of epitopes predicted by the hydropathic profile. The setup was adapted to a novel biochip system based on surface-enhanced absorption for direct CCD-camera based screening. PMID- 12570752 TI - Lead generation--enhancing the success of drug discovery by investing in the hit to lead process. AB - Improving on the poor success rates in the drug discovery industry requires that knowledge-based decisions are made to advance or stop a lead candidate as early as possible in the discovery process. Failure to make such timely decisions on the rigorous selection of lead candidates has costly time and resource implications in downstream drug development. To meet this challenge dedicated 'hit to lead' groups have recently been established in many major pharmaceutical companies, and a key to the success of such groups is establishing a clear consistent process and rigorous metrics for lead quality. The importance of such a "Lead Generation" group within the drug discovery process will be highlighted with the aim of placing a greater level of emphasis in discovering and refining novel lead series with enhanced drug-like properties. This activity is facilitated by the application of productivity enhancing, integrated technologies coupled with the early evaluation of drug-like properties in the lead refinement process to ensure that a balanced activity - properties profile can be attained before committing to a full lead optimisation program. This article will survey the processes and tools employed in the hit to lead process in such a "Lead Generation" group in order to achieve these objectives, emphasising the possible gains in productivity through close, early interactions between chemistry and other expert groups. PMID- 12570754 TI - Solid-phase synthesis of heterocyclic aromates: applicability towards combinatorial chemistry; a review. AB - Because of their biological activity, stability in vivo, the rigid spatial positioning of their substituents, and their synthetic challenges, heterocyclic aromates continue to be of interest to both academic and industrial medicinal chemists. Currently, many drug-like heterocyclic aromates are prepared via solid phase organic chemistry methods. This review examines the applicability of those methods towards combinatorial chemistry with respect to the basic demands of such an approach: 1) synthesis, work-up and subsequent purification should be easily automated enabling the efficient simultaneous synthesis of large numbers of highly pure compounds in a minimum amount of time, 2) large diversity among the ligands to be synthesized, 3) high conversion rates of the individual reaction steps, and 4) the use of commercially available starting materials. Although many methods have been developed for the synthesis of heterocyclic aromates, very few of the available methods enable the synthesis of highly diverse heteroaromatic libraries. PMID- 12570755 TI - Medicinal chemistry of adenosine A1 receptor ligands. AB - In this review the latest developments in ligand design for the adenosine A(1) receptor are summarized. Novel series of agonists and antagonists are discussed, leading to the conclusion that ligands truly selective for the human adenosine A(1) receptor are still scarce. PMID- 12570756 TI - Pharmacology and therapeutic applications of A1 adenosine receptor ligands. AB - Adenosine's diverse physiological functions are mediated by four subtypes of receptors (A(1), A(2A), A(2B) and A(3)). The A(1) adenosine receptor pharmacology and therapeutic application of ligands for this receptor are the subjects of this review. A(1) receptors are present on the surface of cells in organs throughout the body. Actions mediated by A(1) receptors include slowing of heart rate and AV nodal conduction, reduction of atrial contractility, attenuation of the stimulatory actions of catecholamines on beta-adrenergic receptors, reduction of lipolysis in adipose tissue, reduction of urine formation, and inhibition of neuronal activity. Although adenosine analogs with high efficacy, affinity, and selectivity for the A(1) receptor are available, the ubiquitous distribution and wide range of physiological actions mediated by A(1) receptors are obstacles to development of therapeutic agents that activate these receptors. However, it may be possible to exploit the high A(1) "receptor reserve" for some actions of adenosine by use of weak (partial) agonists to target these actions while avoiding others for which receptor reserve is low. The presence of high receptor reserves for the anti-arrhythmic and anti-lipolytic actions of adenosine suggests that partial A(1) agonists could be used as anti-arrhythmic and anti-lipolytic agents. In addition, allosteric enhancers of the binding of adenosine to A(1) receptors could be used therapeutically to potentiate desirable effects of endogenous adenosine. Antagonists of the A(1) receptor can increase urine formation, and because they do not decrease renal blood flow, are particularly useful to maintain glomerular filtration in patients having edema secondary to reduced cardiac function. PMID- 12570757 TI - Medicinal chemistry of adenosine A2A receptor agonists. AB - The search for potent and selective A(2A) adenosine receptor agonists has been particularly fruitful in the early nineties. A series of 2-amino, 2-alkoxy, 2 alkythio-, 2-alkynyl-, and 2-alkenyl-derivatives of adenosine (Ado, 1) and N ethylcarboxamidoadenosine (NECA, 30) have been synthesized and tested mainly on different model of rat A(1) and A(2A) receptor subtypes. From these studies some ligands, such as CGS 21680 (33), HENECA (42), and (S)-PHPNECA (46b), showed to possess high A(2A) affinity combined with good A(2A) vs A(1) selectivity. More detailed characterization of these ligands at the four cloned human adenosine receptor subtypes revealed that none of the prototypical adenosine receptor agonists exhibits at the same time high affinity and selectivity for the human A(2A)AR subtype. Both NECA and CGS 21680, which are avalaible as radioligands for this subtype, have lower affinity at human than at rat receptor. The 2 alkynylNECA derivatives HENECA an PHPNECA showed high affinity also at human A(3) receptors. In particular, (S)-PHPNECA displayed K(i)s in the low nanomolar range at A(1), A(2A), and A(3)subtypes and an EC(50) of 220 nM at human A(2B) receptor. On the other hand, it is now well known that the coronary vasodilation induced by Ado in different species is mediated by activation of A(2A)AR and a compound capable of producing coronary vasodilation through activation of A(2A)AR, but that is devoid of A(1)- and A(1)-agonist activity would have advantage over Ado for use in myocardial perfusion imaging studies. Other potential therapeutic applications of selective A(2A)AR agonists are as anti-aggregatory, anti inflammatory, anti-psychotic, and anti-Huntington's disease agents. This review is aimed at presenting a complete overview of the medicinal chemistry development of A(2A) adenosine receptor agonists and at stressing the strong need for more selective ligands at A(2A) human subtype. PMID- 12570758 TI - Medicinal chemistry of A2A adenosine receptor antagonists. AB - Due to the clearly demonstrated receptor-receptor interaction between adenosine A(2A) and dopamine D(2) receptors in the basal ganglia, the discovery and development of potent and selective A(2A)adenosine receptor antagonists became, in the last ten years, an attractive field of research to discovery new drugs for the treatment of neurodegenerative disorders, such as Parkinsons disease. Different compounds have been deeply investigated as A(2A) adenosine receptor antagonists, which could be classified in two great families: xanthine derivatives and nitrogen poliheterocyclic systems. These studies led to the discovery of some highly potent and selective A(2A) adenosine receptor antagonists such as ZM241385, SCH58261 and some xanthine derivatives (KW6002), which have been used as pharmacological tools for studying this receptor subtype. However, those compounds showed some problems that do not permit their use in clinical studies, such as poor water solubility (SCH58261, and xanthine derivatives) or good affinity for A(2B) adenosine receptor subtype (ZM241385). In the last few years great efforts have been made to overcome these problems, trying to optimize not only the pharmacological profile but also the pharmacokinetic character of this class of compounds. The aim of this report is to briefly summarize the recent progress made in this attractive field of research. PMID- 12570759 TI - Pharmacology of adenosine A2A receptors and therapeutic applications. AB - Adenosine A(2A) receptors were cloned about ten years ago and are known to be well conserved among mammals. Rather selective agonists and antagonists are available. In addition, two different knock-out mice have been prepared and extensively characterized. A(2A) receptors are highly enriched in the basal ganglia and on cells involved in inflammatory reactions. At these sites they are likely to play physiologically important roles. Efforts to develop new therapies based on A(2A) receptors have focused on these topics. However, A(2A) receptors are found on many other cell types and on them as well agonists can exert effect. PMID- 12570760 TI - Medicinal chemistry and pharmacology of A2B adenosine receptors. AB - The low affinity A(2B) adenosine receptor, like any other adenosine receptor subtype, belongs to the super-family of seven transmembrane domain protein coupled receptors (7TMs GPCR) and is classified by the GPCR database in the family of rhodopsin like receptors (Class A of GPCR). It has been cloned from various species, including rat and human, and its sequences are highly similar across species, ranging from 85% identity between human and mouse and 95% identity between rat and mouse. The A(2B)receptors show a ubiquitous distribution, the highest levels are present in cecum, colon and bladder, followed by blood vessels, lung, eye and mast cells. Through A(2B) receptors adenosine seems to cause mast cells degranulation, vasodilation, cardiac fibroblast proliferation, inhibition of Tumor Necrosis Factor (TNF-alpha), increased synthesis of interleukin-6 (IL-6), stimulation of Cl(-) secretion in intestinal epithelia and hepatic glucose production. Hence, A(2B) adenosine receptor agonists could be useful in the treatment of cardiac diseases like hypertension or myocardial infarction and in the management of septic shock, while antagonists may serve as novel drugs for asthma, Alzheimer's disease, cystic fibrosis and type-II diabetes. No potent and selective A(2B) agonists have been reported so far; 5'-N-ethylcarboxamidoadenosine (NECA) is one of the most active. The monosubstitution on N(6)-position of adenosine is well tolerated and that position appears to be a useful site for increasing A(2B) potency. Among substituents in 2-position of adenosine only 1-alkynyl chains are effective for A(2B) potency. In particular, the (S)-2-hydroxypropynyl substituents brought about the highest activity demonstrating that the A(2B) receptors discriminate between (R) and (S) diastereomers. Hence, (S)-2-phenylhydroxypropynylNECA (PHPNECA), with an EC(50) = 0.22 micro M, proved to be the most potent A(2B) agonist reported so far. Classical antagonists for adenosine receptors are alkylxanthines which show considerable potency at A(2B) receptors. Para substituted 1,3-dialkyl-8-phenylxanthines possess high affinity in binding assays; the 3-unsubstituted 1-alkyl analogues resulted more A(2B) selective with the 8-[4-[(N-(2-hydroxyethyl)carboxamidomethyl)oxy]phenyl]-1-propylxanthine (60) showing the highest affinity (K(i) = 1.2 nM) and selectivity (A(1)/A(2B) = 60, A(2A)/A(2B) = 1,790, A(3)/A(2B) = 360). Among non-xanthine derivatives very promising are substituted purines, in which combination of appropriate substituents in 2-, 8-, and 9-position could lead to very potent and selective A(2B) antagonists. PMID- 12570761 TI - Medicinal chemistry of adenosine A3 receptor ligands. AB - A(3) Adenosine receptors (ARs) exhibit large species differences. Potent, selective agonists for rat (e.g. Cl-IB-MECA, 5) and human A(3) ARs (e.g. PENECA, 17, and analogs) have been developed during the past years. Potent, selective antagonists for human A(3) ARs include the imidazopurinones PSB-10 (28) and PSB 11 (29), the pyrazolotriazolopyrimidines MRE-3005F20 (38) and analogs, and the dihydropyridines (e.g. MRS-1334, 50). For rat A(3) ARs only moderately potent antagonists have been identified, such as the pyridine derivative MRS-1523 (51) and the flavonoid MRS-1067 (52), both of which exhibit only a low degree of selectivity versus the other AR subtypes. Selective antagonist radioligands for the human A(3) receptor, [(3)H]MRE-3008F20 and [(3)H]PSB-11, have been prepared, while A(3)-selective agonist radioligands are still lacking. Recent developments also include allosteric modulators, irreversibly binding antagonists, fluorescence-labelled agonists, partial agonists and inverse agonists for A(3)ARs. Site-directed mutagenesis and molecular modeling studies have been performed in order to obtain information about the ligand binding site and the process of receptor activation. A(3)Adenosine receptors have recently attracted considerable interest as novel drug targets. A(3) Agonists may have potential as cardioprotective and cerebroprotective agents, for the treatment of asthma, as antiinflammatory and immunosuppressive agents, and in cancer therapy as cytostatics and chemoprotective compounds. A(3) AR antagonists might be therapeutically useful for the acute treatment of stroke, for glaucoma, and also as antiasthmatic and antiallergic drugs, since A(3)receptors cannot only mediate antiinflammatory, but also proinflammatory responses. The future development of further pharmacological tools, including potent, selective antagonists for rat A(3) receptors and selective agonist radioligands for rat and human receptors will facilitate the evaluation of the (patho)physiological roles of A(3) receptors and the pharmacological potential of their ligands. PMID- 12570762 TI - Pharmacology and therapeutic applications of A3 receptor subtype. AB - The present study summarizes the biological effects elicit upon A(3) adenosine receptor (A(3)AR) activation in normal and tumor cells. Anti-inflamatory response is mediated upon A(3)AR activation in neutrophils, eosinophils and macrophages via direct effect on cell degranulation or the production of anti-inflamatory cytokines. In basophils, which highly express A(3)AR, degranulation and mediator release upon receptor activation lead to pro-inflammatory effects resulting in bronchospasm and asthma. In other normal cells such as cardiomyocytes, neuronal cells and bone marrow cells A(1)AR activation induces cytoprotective effects in vitro. In vivo, A(3)AR agonists act as cardio- and neuroprotective agents and attenuate ischemic damage. Furthermore, agonists to A(3)AR induce granulocyte colony stimulating factor (G-CSF) production and myeloprotective effect in chemotherapy treated mice. Interestingly, A(3)AR agonists inhibit tumor cell growth both in vitro and in vivo through a cytostatic effect mediated via the de regulation of the Wnt signaling pathway. The variety of activities elicit by A(3)AR agonists suggest their potential use as therapeutic agents in inflammation, brain/cardiac ischemia and cancer. Antagonists to A(3)AR may be implemented to the therapy of asthma and additional allergic conditions. PMID- 12570763 TI - Fluoroquinolones: action and resistance. AB - Fluoroquinolones trap gyrase and topoisomerase IV on DNA as ternary complexes that block the movement of replication forks and transcription complexes. Studies with resistant mutants indicate that during complex formation quinolones bind to a surface alpha-helix of the GyrA and ParC proteins. Lethal action is a distinct event that is proposed to arise from release of DNA breaks from the ternary complexes. Many bacterial pathogens are exhibiting resistance due to alterations in drug permeability, drug efflux, gyrase-protecting proteins, and target topoisomerases. When selection of resistant mutants is described in terms of fluoroquinolone concentration, a threshold (mutant prevention concentration, MPC) can be defined for restricting the development of resistance. MPC varies among fluoroquinolones and pathogens; when combined with pharmacokinetics, MPC can be used to identify compounds least likely to enrich mutant subpopulations. Use of suboptimal doses and compounds erodes the efficacy of the class as a whole because resistance to one quinolone reduces susceptibility to others and/or increases the frequency at which resistance develops. When using fluoroquinolones in combination therapy, the development of resistance may be minimized by optimizing regimens for pharmacokinetic overlap. PMID- 12570764 TI - The ATP-binding site of type II topoisomerases as a target for antibacterial drugs. AB - DNA topoisomerases are essential enzymes in all cell types and have been found to be valuable drug targets both for antibacterial and anti-cancer chemotherapy. Type II topoisomerases possess a binding site for ATP, which can be exploited as a target for chemo-therapeutic agents. High-resolution structures of protein fragments containing this site complexed with antibiotics or an ATP analogue have provided vital information for the understanding of the action of existing drugs and for the potential development of novel anti-bacterial agents. In this article we have reviewed the structure and function of the ATPase domain of DNA gyrase (bacterial topoisomerase II), particularly highlighting novel information that has been revealed by structural studies. We discuss the efficacy and mode of action of existing drugs and consider the prospects for the development of novel agents. PMID- 12570765 TI - Non-camptothecin DNA topoisomerase I inhibitors in cancer therapy. AB - Human DNA topoisomerase I is the target of camptothecins, which have been recently introduced in the clinic for cancer chemotherapy. The discovery of novel non-camptothecin inhibitors is facilitated by the availability of biochemical and cellular assays for testing topoisomerase I activity. Among the non-camptothecin inhibitors, the indolocarbazoles (NB-506 and J-107088) are the most advanced in their development, and are in clinical trials. A number of indenoisoquinolines and minor groove binders (benzimidazoles) have been reported recently. Their antitumor activity is promising for further development. The potential binding site(s) of topoisomerase I inhibitors in the enzyme I-DNA complex is discussed. PMID- 12570766 TI - Stabilization of eukaryotic topoisomerase II-DNA cleavage complexes. AB - Topoisomerase II is an essential enzyme that plays critical roles in many DNA processes, including chromosome segregation. In order to carry out its important physiological functions, topoisomerase II creates and rejoins double-stranded breaks in the genetic material. Thus, while the enzyme is necessary for cell survival, it also has the capacity to fragment the genome. Topoisomerase II mediated DNA breaks are sequestered within a covalent enzyme-DNA complex. Normally, these "cleavage complexes" are present at low levels and are tolerated by the cell. However, conditions that significantly increase the physiological concentration or life-time of topoisomerase II-DNA cleavage complexes lead to chromosomal translocations and other mutagenic events, and can induce cell death pathways. The potentially lethal aspect of enzyme mechanism has been exploited by a number of highly successful anticancer agents. Since drugs that increase levels of topoisomerase II-DNA cleavage complexes transform the enzyme into a potent cellular toxin, they are referred to as topoisomerase II "poisons" to distinguish them from compounds that inhibit the catalytic activity of the enzyme. Recent evidence indicates that many DNA lesions also act as topoisomerase II poisons. This finding has provided tremendous insight into enzyme and drug action and raises important questions regarding the physiological interactions of topoisomerase II with DNA damage. Since the DNA cleavage and ligation reactions of topoisomerase II are fundamental to its physiological and pharmacological functions, this review will focus on how the enzyme cuts and rejoins the double helix and how these reactions are altered by topoisomerase II poisons. PMID- 12570767 TI - Dual topoisomerase I/II inhibitors in cancer therapy. AB - While the majority of topoisomerase (topo) inhibitors show selectivity against either topo I or topo II, a small class of compounds can act against both enzymes. These can be divided into three classes. The first and largest class comprise drugs that bind to DNA by intercalation and include the clinically evaluated acridine DACA, the benzopyridoindole intoplicine, the indenoquinolinone TAS-103, the benzophenazine XR11576, and the pyrazoloacridine NSC 366140. The second category comprises hybrid molecules, prepared by physically linking separate inhibitors of topo I and topo II, or by linking pure topo inhibitors to other DNA-interactive carriers. While several derivatives (e.g., camptothecin epipodophyllotoxin and ellipticine-distamycin hybrids) have been prepared, there have been no detailed studies. The third category are less well defined as a structural class, but apparently recognize structural motifs that are present in both topo I and II enzymes. These include a series of benzoisoquinolinium quaternary salts such as NK 109, and more interestingly modified versions of classical topo I or topo II inhibitors; e.g., the modified camptothecin BN 80927 and the modified epipodophyllotoxin tafluposide (F-11782). There is as yet no detailed understanding of the factors that result in selective or dual inhibition, but structure-activity studies in several classes show that structural changes can influence topo I/II selectivity. DNA intercalation mode also appears to play a part. The basis for the high antitumor activity of some topo inhibitors is not yet understood but may depend on the complex pattern of activities that include both inhibition and poisoning of the two enzymes. PMID- 12570768 TI - Biodynamic constituents in the Mexican morning glories: purgative remedies transcending boundaries. AB - Jalap, a pre-Hispanic herbal remedy still considered a useful laxative, is an ingredient in some over-the-counter products sold by herbalists in contemporary Mexico. The purgative crude drugs are prepared from the roots of several morning glories species which all have been identified as members of the Ipomoea genus (Convolvulaceae). Their incorporation as therapeutical agents into Europe is an outstanding example of the assimilation of botanical drugs from the Americas as substitutes for traditional Old World remedies. Phytochemical investigations on the resin glycosides, main constituents of these drugs, were initiated during the second half of the XIX century; however, the structures of their active ingredients had remained poorly known, and still are for some members of these purgative root species. Modern analytical techniques with higher resolution capabilities (HPLC) for the isolation of the active principles of these crude drugs used in conjunction with powerful spectroscopic methods (high field NMR) have facilitated the investigation of these relevant, to the herbal product market, convolvulaceous species during the last decade. The advantages and limitations of theses techniques will be discussed. This review will also describe the ethnobotanical information associated with the Mexican morning glory species and how the traditional usages of these plants have played an important role in the selection of these materials for chemical studies. Little is as yet known about either the mechanism of purge action caused by the resin glycosides or the ecological significance of these same compounds for the producing plants. Over the five centuries of Mexican herbal medicine, one hundred years of phytochemistry has only partially elucidated the active ingredients of the jalap roots but has exemplified how to further contemporary drug discoveries through the investigation of those plants traditionally held to be economically and medicinally important in developing countries. PMID- 12570769 TI - Traditionally-used antimalarials from the Meliaceae. AB - A quantitative ethnobotanical approach to antimalarial drug discovery led to the identification of Lansium domesticum Corr. Ser. (Meliaceae) as an important antimalarial used by Kenyah Dyak healers in Indonesian Borneo. Triterpenoid lansiolides with antimalarial activity were isolated from the bark and shown to have activity in both in vitro bioassays with Plasmodium falciparum, and in mice infected with P. berghei. A survey of African and tropical American Meliaceae led to further development of the limonoid gedunin from the traditionally used medicinal plants, tropical cedar, Cedrela odorata L., and neem, Azadirachta indica A. Juss. Gedunin has significant in vitro activity but initially showed poor in vivo activity. In vivo activity was improved by (1) incorporation into an easy to absorb suspension, (2) preparation of a more stable compound, 7 methoxygedunin; and (3) synergism with dillapiol, a cytochrome P450 3A4 inhibitor. The results show the potential for both antimalarial drug and phytomedicine development from traditionally used plants. PMID- 12570770 TI - Ethnobotany and natural products: the search for new molecules, new treatments of old diseases or a better understanding of indigenous cultures? AB - Results of various projects on Mexican Indian ethnobotany and some of the subsequent pharmacological and phytochemical studies are summarised focusing both on chemical-pharmacological as well as anthropological (ethnopharmacological) aspects of our research. We have identified taste and smell properties of medicinal (vs. non-medicinal) plants as important indigenous selection criteria. There exist well-defined criteria specific for each culture, which lead to the selection of a plant as a medicine. This field research has also formed a basis for studies on bioactive natural products from selected species. The bark of Guazuma ulmifolia showed antisecretory activity (cholera toxin-induced chloride secretion in rabbit distal colon in an USSING chamber). Active constituents are procyanidins with a polymerisation degree of eight or higher. Byrsonima crassifolia yielded proanthocyanidins with (+) epicatechin units and Baccharis conferta showed a dose-dependent antispasmodic effect with the effect being particularly strong in flavonoid-rich fractions. Our ethnopharmacological research led to the identification of sesquiterpene lactones (SLs) like parthenolide as potent and relatively specific inhibitors of the transcription factor NF-kappaB, an important mediator of the inflammatory process. The inhibitory effect of SLs is very strongly enhanced by the presence of such groups as the isoprenoid ring system, a lactone ring containing a conjugated exomethylene group (alpha-methylene-gamma-lactone) and an alpha,beta-unsaturated cyclopentenone or a conjugated ester moieties. Our work also elucidated the NF kappaB inhibiting activity of the photosensitiser phaeophorbide A from Solanum diflorum (Solanaceae) in PMA induced HeLa cells. Hyptis verticillata yielded a series of lignans as well as sideritoflavone, rosmarinic acid and (R)-5 hydroxypyrrolidin-2-one and is rich in essential oil (rich in alpha-pinene, beta pinene and thymol). Other species investigated include Begonia heracleifolia, Crossopetalum gaumerii, Epaltes mexicana, Pluchea symphytifolia and Xanthosoma robustum. PMID- 12570771 TI - Plant-derived terpenoids and analogues as anti-HIV agents. AB - The plant-derived terpenoids and analogues are reviewed with respect to their anti-HIV activity, structure-activity relationships, and mechanism of action. The active compounds include diterpenoid lactones, phenolic diterpenes, atisane and kaurane diterpenes, phorbol diterpenes, triterpene glycosides, friedelane triterpenes, taraxerane triterpenes, ursane triterpenes, lanostane triterpene, lupane triterpenes, seco-ring A triterpenes, degraded triterpenes, and cucurbitacin triterpenes. Positive new leads for drug development will be highlighted. PMID- 12570772 TI - Terpenoids: sources, structure elucidation and therapeutic potential in inflammation. AB - Natural products research has lately undergone exponential growth owing to advances in isolation techniques and in synthetic methods design, as well as for the identification of a wide range of biological properties exhibited by these compounds. In the present review, general remarks on the chemical features, biosynthetic pathways, isolation and structure elucidation of terpenoids are briefly discussed. In addition to this, recent work done on anti-inflammatory terpenoids (diterpenoids, triterpenoids and sesquiterpene lactones) with special emphasis on the last new molecular targets evaluated is presented. PMID- 12570773 TI - Hypericum perforatum, a source of neuroactive lead structures. AB - Although St. John's wort has been known for thousands of years and has been used for a variety of medicinal purposes, understanding of its activity and mechanisms of action is relatively new and not well understood. While researchers originally thought the naphthodianthrone hypericin was responsible for Hypericum's antidepressant activity, it is now believed some other compound or a combination of constituents exerts their antidepressant activity on the body. Hypericum is unique in that it seems to impact all known neurotransmitters at some level, directly, or indirectly through receptor sensitivity and regulation. There has been a proliferation of clinical studies on Hypericum in the last ten years, and even though some of these studies might be methodologically flawed, the preponderance of the evidence proves Hypericum to be beneficial for the treatment of mild-to-moderate depression, with a very favorable side effect profile. One clinical trial carried out using two extracts with different hyperforin content indicate this constituent as (one of) the main active principle responsible for the antidepressant activity. PMID- 12570774 TI - Novel diarylheptanoids of Alpinia blepharocalyx. AB - The seeds of Alpinia blepharocalyx K. Schum. (Zingiberaceae) is used in Chinese traditional medicine for the treatment of stomach disorders. From the ether fraction of a 95% ethanolic extract, which showed hepatoprotective and antiproliferative activities, we isolated 16 novel diarylheptanoids bearing a chalcone or a flavanone moiety [calyxins A-H; epicalyxins B-D, G, and H; 6 hydroxycalyxin F; and blepharocalyxins A and B] together with seven known compounds, while the residual fraction of the ethanolic extract gave 32 novel diarylheptanoids namely, calyxins A, E-G, and I-M; epicalyxins B, F, I-K, and M; deoxycalyxin A; blepharocalyxins C-E; neocalyxins A and B; (3S,5S)- and (3S,5R)-3 hydroxy-1-(4-hydroxyphenyl)-5-methoxy-7-phenyl-6E-heptene, (3S,5S)- and (3S,5R)-3 hydroxy-1-(4-hydroxyphenyl)-5-ethoxy-7-phenyl-6E-heptene, (3S)-3-methoxy-1,7 bis(4-hydroxyphenyl)-6E-hepten-5-one, 1,7-bis(4-hydroxyphenyl)-hepta-4E,6E-dien-3 one, (3S,7R)-5,6-dehydro-1,7-bis(4-hydroxy-phenyl)-4"-de-O-methyl-centrolobine, (3S,5S,6S,7R)-5,6-dihydroxy-1,7-bis(4-hydroxyphenyl)-4"-de-O-me-thylcentrolobine, (3S,5R,6S,7R)- and (3S,5S,6R,7R)-5,6-dihydroxy-1,7-bis(4-hydroxyphenyl)-4"-de-O methyl-centrolobine, 1,2- dihydro-bis(de-O-methyl)curcumin, and (3S,7S)-5,6 dehydro-4"-de-O-methylcentrolobine, and one known diarylheptanoid [(3S,5S)-3,5 dihydroxy-1,7-bis(4-hydroxyphenyl)heptane] together with 12 other known phenolic compounds. Moreover, in vitro NO inhibitory and antiproliferative activities of the isolated compounds were also tested and the active constituents identified. PMID- 12570775 TI - Chemical constituents and biological activities of Vietnamese medicinal plants. AB - Every country has its own traditional medicine system originated from ancient times. Vietnam, a Southeast Asian country, has a unique traditional medicine system. Several medicinal plants are still used by different ethnic societies of Vietnam for treating several diseases due to their easy availability. Combretum quadrangulare, Panax vietnamensis (Vietnamese Ginseng), Dracaena angustifolia (Nam Ginseng) and Orthosiphon stamineus are among the popular medicinal plants used in Vietnamese traditional medicine for curing various diseases and to improve physical strength. In this review article, we discuss the chemical analysis as well as biological activities of these Vietnamese medicinal plants in order to establish the relation between their therapeutic uses and scientific findings. PMID- 12570776 TI - Design of small molecule libraries for NMR screening and other applications in drug discovery. AB - There are conceptual differences between high-throughput screening (HTS) and fragment-based screening by NMR. The number of compounds in libraries for NMR screening may be significantly smaller than those used for HTS. Because one relies on a small library its design is significantly important and is the object of this article. A short introduction on fragment-based NMR screening approaches will be provided. Although there are currently very few reports describing the design of libraries of small molecules for NMR screening, aspects of the question of how to compile diverse collections of small molecular fragments useful for drug design were previously addressed for the purposes of combinatorial library design and de novo drug design. As these disciplines are highly interrelated and are applied in an interconnected manner with NMR screening within the drug discovery process, a review of combinatorial library design and especially the building block or fragment selection strategies applied for combinatorial library design and de novo design is well suited to reveal fundamental strategies and potential techniques for the design of NMR screening libraries. This section will be rounded off by a report on hands-on-experience with the design of the Novartis second-site NMR screening library and practical considerations for the design of compound mixtures. Rather than providing an exact protocol general guidelines will be indicated. PMID- 12570777 TI - Automation of biomolecular NMR screening. AB - For NMR based screening we review equipment needed for automated preparation of samples and acquisition of a large number of data sets. Hardware connecting lab bench and NMR spectrometer is described. We focus on software used for automated calculation of the similarity between spectra - a prerequisite for the identification of test compounds interacting with a target-molecule. PMID- 12570778 TI - Second-site NMR screening and linker design. AB - One of the prime merits of NMR as a tool for lead finding in drug discovery research is its sensitivity and robustness to detect weak protein-ligand interactions. This sensitivity allows to build up ligands for a given target in a modular way, by a fragment-based approach. In this approach, two ligands are seperately identified which bind to the target protein generally weakly, but at adjacent binding sites. In a next step, they are chemically linked to produce a high-affinity ligand. This review discusses methods to detect "second-site" ligands that bind to a protein in the presence of a "first-site" ligand, and methods to elucidate structural details on the spatial orientation of both ligands, so that chemical linkage is based on a large piece of experimental information. Published examples from second-site screening and linker design are summarized, and are complemented by previously unpublished in-house examples. PMID- 12570779 TI - Application of NMR screening in drug discovery. AB - The application of NMR screening in drug discovery has recently attained heightened importance throughout the pharmaceutical industry. NMR screening can be applied at various points in a drug discovery program, ranging from very early in the program, when new targets can be screened long before an HTS enzymatic assay is developed, to later in the program, as in the case where no useful hits have been detected by HTS using biological assays. The binders determined in primary NMR screens are used to guide secondary screens, which can be either completely NMR driven or use NMR in combination with other biophysical techniques. In this review we briefly discuss the methods and techniques used in NMR screening. Then, we describe in detail the NMR screening strategies and their applications to specific targets, including successful examples from actual drug design programs at our own and other pharmaceutical companies. PMID- 12570780 TI - Why are glycoproteins modified by poly-N-acetyllactosamine glyco-conjugates? AB - Poly-N-acetyllactosamine structures occur in mammalian glycoproteins in both N- and O-linked glycans. They represent a backbone for additional modifications by fucosyltransferases, sialyltransferases and sulfotransferases. These glycans have been suggested to be involved in biospecific interactions with selectins and other glycan-binding proteins. Moreover, the poly-N-acetyllactosamine chains in N glycans have been found to promote tumor progression and metastasis. Poly-N acetyllactosamine chains are synthesized by repeated alternating additions of N acetylglucosamine and galactose, catalyzed by beta-1,3-N acetylglucosaminyltransferases (poly-N-acetyllactosamine synthase) and beta-1,4 galactosyltransferases. This review describes the poly-N-acetyllactosamine assembling machinery and focuses on recent advances in the molecular cloning and characterization of poly-N-acetyllactosamine synthase gene families. Recent progress in revealing the biological functions of poly-N-acetyllactosamine structures by various approaches in vitro and in vivo using different model systems has also been summarized. PMID- 12570781 TI - Mapping protein: carbohydrate interactions. AB - Many biologically important interactions occur between proteins and carbohydrates. The examination of these interactions at the atomic level is critical not only in understanding the nature of these interactions and their biological role, but also in the design of effective modulators of these interactions. While experimentally obtained structural information is preferred, quite often this information is unavailable. In order to address this, several methods have been developed to probe the interactions between protein and carbohydrate in the absence of structural data. These methods map the interactions between protein and carbohydrate, and identify the groups involved, both at the carbohydrate and protein level. Here, we review these developments, and examine the strengths, weaknesses, and pitfalls of these methods. PMID- 12570782 TI - Bacterial beta-ketoacyl-acyl carrier protein synthases as targets for antibacterial agents. AB - As a result of increasing drug resistance in pathogenic bacteria, there is a critical need for novel broad-spectrum antibacterial agents. As fatty acid synthesis (FAS) in bacteria is an essential process for cell survival, the enzymes involved in the FAS pathway have emerged as promising targets for antimicrobial agents. Several lines of evidence have indicated that bacterial condensing enzymes are central to the initiation and elongation steps in bacterial fatty acid synthesis and play a pivotal role in the regulation of the entire fatty acid synthesis pathway. beta-ketoacyl-acyl carrier protein (ACP) synthases (KAS) from various bacterial species have been cloned, expressed and purified in large quantities for detailed enzymological, structural and screening studies. Availability of purified KAS from a variety of bacteria, along with a combination of techniques, including combinatorial chemistry, high-throughput screening, and rational drug design based on crystal structures, will undoubtedly aid in the discovery and development of much needed potent and broad-spectrum antibacterial agents. In this review we summarize the biochemical, biophysical and inhibition properties of beta-ketoacyl-ACP synthases from a variety of bacterial species. PMID- 12570783 TI - Neuropeptide conversion to bioactive fragments--an important pathway in neuromodulation. AB - Biosynthetic pathways for the formation of neuroactive peptides and the processes for their inactivation include several enzymatic steps. In addition to enzymatic processing and degradation, several neuropeptides have been shown to undergo enzymatic conversion to fragments with retained or modified biological activity. This has most clearly been demonstrated for e.g. opioid peptides, tachykinins, calcitonin gene-related peptide (CGRP) as well as for peptides belonging to the renin-angiotensin system. Sometimes the released fragment shares the activity of the parent compound. However, in many cases the conversion reaction is linked to a change in the receptor activation profile, i.e. the generated fragment acts on and stimulates a receptor not recognized by the parent peptide. This review will describe the characteristics of certain neuropeptide fragments having the ability to modify the biological action of the peptide from which they are derived. Focus will be directed to the tachykinins, the opioid peptides, angiotensins as well as to CGRP, bradykinin and nociceptin. The kappa opioid receptor selective opioid peptide, dynorphin, recognized for its ability to produce dysphoria, is converted to the delta opioid receptor agonist Leu-enkephalin, with euphoric properties. The tachykinins, typified by substance P (SP), is converted to the bioactive fragment SP(1-7), a heptapeptide mimicking some but opposing other effects of the parent peptide. The bioactive angiotensin II, known to bind to and stimulate the AT-1 and AT-2 receptors, is converted to angiotensin IV (i.e. angiotensin 3-8) with preference for the AT-4 sites or to angiotensin (1-7), not recognized by any of these receptors. Both angiotensin IV and angiotensin (1-7) are biologically active. For example angiotensin (1-7) retains some of the actions ascribed for angiotensin II but is shown to counteract others. Thus, it is obvious that the activity of many neuroactive peptides is modulated by bioactive fragments, which are formed by the action of a variety of peptidases. This phenomenon appears to represent an important regulatory mechanism that modulates many neuropeptide systems but is generally not acknowledged. PMID- 12570784 TI - Hormonal control of the neuropeptide Y system. AB - Neuropeptide Y (NPY) and the related receptors represent a widely diffused system that is involved in the regulation of multiple biological functions. NPY, a 36 aminoacid peptide expressed in several areas of the nervous system, is a pleiotropic factor participating to the control of some physiological processes, such as cognitive functions, eating behavior, circadian rhythms, neuroendocrine mechanisms, reproductive and cardiovascular functions. NPY acts through a series of G-protein-associated membrane receptors (NPY-Rs), characterized by different tissue distribution and affinity for the ligand. The expression and secretion of NPY and the expression of NPY-R isoforms are controlled by a very wide range of agents, acting in an endocrine and/or paracrine fashion. NPY and NPY-Rs appear to be strongly involved in the control of eating behavior; their expression is modulated by changes of food intake and energy balance and is disrupted in several animal models of obesity and diabetes. Moreover, the hypothalamic NPY system appears to integrate signals of energy balance in the modulation of the reproductive axis. Agents that stimulate their expression include activators of intracellular signalling pathways (protein kinase A and C), classical neurotransmitters, steroid and peptide hormones and growth factors, while other agents (leptin, insulin and retinoic acid) have been shown to be inhibitory. Interestingly, some agents, like retinoic acid, have been shown to modulate the expression of both NPY and NPY-Rs in the same direction, thus providing a fine mechanism for the tuning of the system. The regulation of NPY/NPY-R expression and function appears to be part of a complex system controlling multiple physiological functions, and its disruption might be relevant in the pathophysiology of disease states such as obesity. PMID- 12570785 TI - Structural/functional aspects of ES-62--a secreted immunomodulatory phosphorylcholine-containing filarial nematode glycoprotein. AB - ES-62 is a major secreted glycoprotein of the rodent filarial nematode Acanthocheilonema viteae and homologue of molecules found in filarial nematodes which parasitise humans. The molecule consists of a tetramer of apparently identical monomers of ~62 kDa which we have shown by sedimentation equilibrium analytical ultracentrifugation to strongly associate. ES-62 is one of several filarial nematode proteins to contain the unusual post-translational modification of phosphorylcholine (PC) addition. Specifically, we have found that PC is attached to one of three distinct N-type glycans we have characterised on the molecule. The amino acid sequence of ES-62 shows 37-39% identity with a family of 6 other proteins, some of which have been predicted to be amino- or carboxy peptidases. We have also found that ES-62 is able to interact with a number of cells of the immune system, specifically B- and T-lymphocytes, macrophages and dendritic cells. Lymphocytes exposed to ES-62 in vitro or in vivo are less able to proliferate in response to ligation via the antigen receptor. Peritoneal macrophages pre-exposed to the molecule are less able to produce the cytokines IL 12, IL-6 and TNF-alpha following subsequent incubation with the classical stimulators IFNgamma and LPS. Dendritic cells allowed to mature in the presence of ES-62 acquire a phenotype, which allows them to induce anti-inflammatory "TH2 type" responses. With respect to immunomodulation, the PC moiety of the parasite molecule appears to be predominantly responsible for the effects on lymphocyte proliferation at least and we have also found that its removal converts the murine IgG antibody response to ES-62 from solely IgG1 to mixed IgG1/IgG2a. ES-62 appears to interact with cells of the immune system in a PC-dependent manner and, at least in part, via a molecule of ~82 kDa. Studies of the interaction in lymphocytes show that it is associated with activation of certain signal transduction molecules including a number of protein tyrosine kinases and mitogen activated protein kinases (MAPkinases). Although such activation is insufficient to induce proliferation, it serves to almost completely desensitise the cells to antigen-receptor ligation-induced activation of the phosphoinositide 3-kinase (PI 3-kinase) and Ras/MAPkinase pathways, events critical for lymphocyte proliferation. Such desensitisation reflects ES-62-primed recruitment of a number of negative regulators of these pathways, such as the phosphatases SHP-1 and Pac 1. PMID- 12570786 TI - Rapid translation system (RTS): a promising alternative for recombinant protein production. AB - Rapid Translation System (RTS) is a cell-free protein production system employing an enhanced Escherichia coli lysate to perform coupled in vitro transcription translation reactions. A continuous supply of energy substrates, nucleotides and amino acids combined with the removal of by-products guarantees a high yield of protein production. The gene to express is either cloned into a plasmid vector or introduced as a PCR product amenable to automation. The main property of this alternative system to cellular expression systems is its open design allowing direct manipulation of the reaction conditions and applications that are impossible or difficult in cell-based systems. RTS offers new promising possibilities in the postgenomic era. PMID- 12570787 TI - Properties and distribution of angiotensin I converting enzyme. AB - This review summarizes some basic properties and distribution of angiotensin I converting enzyme (ACE). ACE is one of several biologically important ectoproteins that exists in both membrane-bound and soluble forms. Localized on the surface of various cells, ACE is inserted at the cell membrane via its carboxyl terminus. Human plasma ACE originates from endothelial cells while other body fluids may contain ACE that originates from epithelial, endothelial or germinal cells. The two isoforms of ACE, the two-domain somatic form and the single domain germinal form, convert angiotensin I to angiotensin II, and metabolize kinins and many other biologically active peptides, including substance P, chemotactic peptide and opioid peptides. The broad spectrum of substrates for ACE and its wide distribution throughout the body indicates that this enzyme, in addition to an important role in cardiovascular homeostasis, may be involved in additional physiologic processes such as neovascularization, fertilization, atherosclerosis, kidney and lung fibrosis, myocardial hypertrophy, inflammation and wound healing. Future research should explore the possible functions of tissue ACE and its systemic role as a pressor agent. ACE inhibitors have achieved widespread use in the treatment of hypertension and the protection of end-organ damage in cardiovascular and renal diseases. Potential problems related to side effects and compliance of such therapy need to be addressed. A safer way of producing therapeutic effects is promised by the delivery of the ACE antisense sequences by a vector producing a permanent inhibition of ACE and long term control of blood pressure in hypertensive patients. PMID- 12570788 TI - Blockade of apoptosis by ACE inhibitors and angiotensin receptor antagonists. AB - Inhibitors of angiotensin converting enzyme (ACE-Is) and angiotensin (ANG) receptor antagonists were originally developed to aid in the management of hypertension. As the use of these agents was extended to the management of heart failure and other cardiovascular diseases, studies of tissue remodeling suggested that blockade of ANGII function might play a role in the regulation of cell death by apoptosis. Experiments with cultured cells confirmed that ANGII is an inducer of apoptosis in well differentiated cell types isolated from the heart, kidneys, lungs and other organs. More recent evidence with animal models strongly suggests that ACE-Is and ANG receptor antagonists, in addition to affecting hemodynamics, also influence disease progression through direct inhibition of ANG-induced apoptosis. This manuscript reviews the evidence supporting this view, discusses its potential relevance to disease pathogenesis and offers new hypotheses regarding novel uses of ACE-Is and ANG receptor antagonists in the control of cell death. PMID- 12570789 TI - The pulmonary renin-angiotensin system. AB - The circulating renin-angiotensin system (RAS) has a well-described role in circulatory homeostasis. Recently, local tissue-based RAS have also been described which appear to play a key role in the injury/repair response. The expression of RAS components and the elevation of angiotensin converting enzyme in a number of interstitial lung diseases suggests the existence of a pulmonary RAS and that angiotensin II could mediate, at least in part, the response to lung injury. Activation of a local RAS within the pulmonary circulation and lung parenchyma could influence the pathogenesis of lung injury via a number of mechanisms including an increase in vascular permeability, vascular tone and fibroblast activity, and by reducing alveolar epithelial cell survival. The ability of both ACE inhibitors and angiotensin II receptor antagonists to attenuate experimental lung injury further supports a role for RAS activation and suggests these agents may be useful in the treatment of diffuse parenchymal lung disease. However, further studies are required to delineate the cell types responsible for RAS component expression in the lung and also to identify the key effector molecules of this system. The presence of common polymorphisms in RAS genes and their study in relation to specific physiological phenotypes will aid both our understanding of the role of RAS in the lung and also aid the targeting of future therapies. PMID- 12570790 TI - Angiotensin converting enzyme inhibitors and angiotensin II receptor antagonists in experimental myocarditis. AB - Myocarditis is a disease whose pathogenesis is not completely understood and whose prevalence is likely underestimated. Individuals afflicted with this condition may be treated with agents that relieve symptoms arising from inflammation and concurrent cellular damage. One class of drugs commonly used in the treatment of myocarditis includes the angiotensin converting enzyme inhibitors, such as captopril, enalapril and lisinopril, and the angiotensin Pi receptor antagonists, such as L-158,809 and losartan. The effects of these drugs on cardiomyopathy have been studied using a variety of animal models of heart failure and hypertension. However, less research has been done in the area of animal models of frank myocarditis. Here we review the use of angiotensin converting enzyme inhibitors and angiotensin Pi receptor antagonists in animal models of myocarditis. We extend the implications of that published work by correlation with results from studies of other disease models and in vitro experiments that highlight the immunomodulatory potential of these compounds. The literature strongly suggests that aggressive therapy employing angiotensin converting enzyme inhibition and/or blockade of angiotensin Pi receptors is beneficial. Treatment is useful not only for reducing complications associated with myocarditis, but also for downregulating the potential autoimmune component of disease without increasing the levels of the infectious agent that may initiate the myocarditis. PMID- 12570791 TI - ACE inhibitors and AII receptor antagonists in the treatment and prevention of bone marrow transplant nephropathy. AB - Radiation nephropathy has emerged as a major complication of bone marrow transplantation (BMT) when total body irradiation (TBI) is used as part of the regimen. Classically, radiation nephropathy has been assumed to be inevitable, progressive, and untreatable. However, in the early 1990's, it was demonstrated that experimental radiation nephropathy could be treated with a thiol-containing ACE inhibitor, captopril. Further studies showed that enalapril (a non-thiol ACE inhibitor) was also effective in the treatment of experimental radiation nephropathy, as was an AII receptor antagonist. Studies also showed that ACE inhibitors and AII receptor antagonists were effective in the prophylaxis of radiation nephropathy. Interestingly, other types of antihypertensive drugs were ineffective in prophylaxis, but brief use of a high-salt diet in the immediate post-irradiation period decreased renal injury. A placebo-controlled trial of captopril to prevent BMT nephropathy in adults is now underway. Since excess activity of the renin-angiotensin system (RAS) causes hypertension, and hypertension is a major feature of radiation nephropathy; an explanation for the efficacy of RAS antagonism in the prophylaxis of radiation nephropathy would be that radiation leads to RAS activation. However, current studies favor an alternative explanation, namely that the normal activity of the RAS is deleterious in the presence of radiation injury. On-going studies suggest that efficacy of RAS antagonists may involve interactions with a radiation-induced decrease in renal nitric oxide activity or with radiation-induced tubular cell proliferation. We hypothesize that while prevention (prophylaxis) of radiation nephropathy with ACE inhibitors, AII receptor antagonists, or a high-salt diet work by suppression of the RAS, the efficacy of ACE inhibitors and AII receptor antagonists in treatment of established radiation nephropathy depends on blood pressure control. PMID- 12570792 TI - Cytostatic properties of some angiotensin I converting enzyme inhibitors and of angiotensin II type I receptor antagonists. AB - Angiotensin converting enzyme (ACE) inhibitors and angiotensin II (AII) type 1 receptor antagonists have strong cytostatic properties on in vitro cultures of many normal and neoplastic cells. They are effective, in particular, in reducing the growth of human lung fibroblasts, renal canine epithelial cells, bovine adrenal endothelial cells, simian T lymphocytes, and of neoplastic cell lines derived from human neuroblastomas, a ductal pancreatic carcinoma of the Syrian hamsters, human salivary glands adenocarcinomas, and two lines of human breast adenocarcinomas. ACE inhibitors are also effective in protecting lungs, kidneys and bladders from the development of nephropathy, pneumopathy, cystitis, and eventually fibrosis in different models of organ-induced damage such as exposure to radiation, chronic hypoxia, administration of the alkaloid monocrotaline or bladder ligation. ACE inhibitors and AII type 1 receptor antagonists are also effective in reducing excessive vascular neoformation in a model of injury to the cornea of rats and rabbits, and in controlling the excessive angiogenesis observed in the Solt-Farber model of experimentally induced hepatoma, in methylcholantrene or radiation-induced fibrosarcomas, in radiation-induced squamous cell carcinomas and in the MA-16 viral-induced mammary carcinoma of the mouse. Captopril was, in addition, effective in controlling tumor growth in a case of Kaposi's sarcoma in humans. The inhibition of AII synthesis and/or its blockade by AII receptors is likely to be an important mechanism for this cytostatic action. The mitogenic effect of AII is well established and a reduction of AII synthesis may well explain cell and neoplasm delayed growth. Moreover, AII regulates and enhances the activity of several growth factors including transforming growth factor B (TGFB) and smooth muscle actin (SMA); and many of these factors are reduced in tissues of animals treated with ACE inhibitors and AII type 1 receptor antagonists. These processes seem to be particularly relevant in the control of fibroblast growth and in the control of the ensuing fibrosis. The ACE inhibitors containing a sulphydril (SH) or thiol radical in their moiety (Captopril and CL242817) seemed to be more effective in controlling fibrosis and the growth of some neoplastic cells than those ACE inhibitors without this thiol radical in their structure, even if the second group of these drugs show in vitro a stronger inhibitory effect on converting enzyme activity. Pharmacologically it is known that ACE inhibitors containing a thiol radical also have antioxidant properties and they are efficient in controlling metalloproteinase action. However, although these additional properties are pharmacologically relevant, the blockade of AII synthesis plays an essential role in the cytostatic activity of these two categories of drugs. These observations underline that in addition to the beneficial effect of these drugs on the cardiovascular system, new potential applications are opening for their wider deployment. PMID- 12570793 TI - Angiotensin-converting enzyme inhibitors: mechanisms of action and implications in anesthesia practice. AB - This review summarizes physiology of circulating and local renin-angiotensin system (RAS), enzymatic properties and mechanism of action of angiotensin I converting enzyme inhibitors (ACEIs) on RAS, and implications of ACEIs in anesthetic management of patients treated with these drugs. ACEIs, through their effect on RAS, may improve cardiovascular functions, pulmonary dynamics, and body fluid homeostasis. Thus, ACEIs have become an integral part of management of patients with hypertension, congestive heart failure (CHF) and chronic renal disease. ACEIs, due to differences in their chemical structure, exert different pharmacological actions and can have protective or occasional damaging effects on different organs. The anesthesiologists are commonly involved in the management of patients treated with ACEIs. Thus, the role of ACEIs and their possible interaction with anesthetic agents must be an integral part of clinical decision making during anesthesia Hemodynamic variation during anesthesia is mainly related to specific effects of anesthetic agents on sympathetic nervous system. Those with preoperative fasting, volume depletion and extended sympathetic blockade can have reduced vascular capacitance resulting in decreased venous return, reduced cardiac output and severe arterial hypotension. Angiotensin II (ANG2) a potent vasoconstrictor may counterbalance such hypotensive effect. During ACE inhibition ANG2 cannot counterbalance this hypotension. Thus, induction of anesthesia may cause severe hypotension in hypovolemic patients specifically in those receiving diuretics as a complement to ACEIs. Recent advances in RAS and the pharmacology of ACEIs have identified some predisposing factors and risks associated with anesthesia in patients treated with ACEIs. Practitioners should be vigilant, and readily have vasopressors, necessary fluids and other resuscitative measures for treatment of unexpected hemodynamic instability during anesthesia and surgery. PMID- 12570794 TI - Aprotinin: a serine protease inhibitor with therapeutic actions: its interaction with ACE inhibitors. AB - Aprotinin is an important member of a family of related protease inhibitors and has many clinically beneficial activities. These inhibitors have multiple functions, but not all of them are mediated by enzyme inhibition. Aprotinin has complex effects on many homeostatic functions including coagulation, platelet function and inflammation. It also has complex interactions with other drug therapies including angiotensin-converting enzyme inhibitors. Since patients with cardiovascular diseases are treated frequently with angiotensin-converting enzyme inhibitors and also often need cardiopulmonary bypass surgery and receive aprotinin, these interactions are potentially significant but often overlooked. Aprotinin is currently used to reduce the amount of transfused homologous blood (during cardiopulmonary bypass surgery) and thus, the risks associated with homologous blood transfusion. Aprotinin also has potential uses in acute pancreatitis, carcinoid tumors, sepsis, and other clinical situations. Future research will provide a definitive answer for the need to employ this inhibitor therapeutically in these situations. Aprotinin also has some potentially adverse effects in the kidney in special circumstances. For example, the use of aprotinin in diabetic patients may be related with an increased risk for renal dysfunction. It has also been associated with thrombosis, inadequate coagulation, and allergic reactions. In balance, the available information indicates that the advantages of its application outweigh its disadvantages in most patients. PMID- 12570795 TI - Neuronal high-affinity sodium-dependent glutamate transporters (EAATs): targets for the development of novel therapeutics against neurodegenerative diseases. AB - L-Glutamate is the major excitatory neurotransmitter in mammalian central nervous system, and excitatory amino acid transporters (EAATs) are essential for terminating synaptic excitation and for maintaining extracellular glutamate concentration below toxic levels. Although the structure of these channel-like proteins has not been yet reported, their membrane topology has been hypothesised based on biochemical and protein sequence analyses. In the case of an inadequate clearance from synaptic cleft and from the extrasynaptic space, glutamate behaves as a potent neurotoxin, and it may be related to several neurodegenerative pathologies including epilepsy, ischemia, amyotrophic lateral sclerosis, and Alzheimer disease. The recent boom of glutamate is demonstrated by the enormous amount of publications dealing with the function of glutamate, with its role on modulation of synaptic transmission throughout the brain, mainly focusing: i). on the structure of its receptors, ii). on molecular biology and pharmacology of Glu transporters, and iii). on the role of glutamate uptake and reversal uptake in several neuropathologies. This review will deal with the recent and most interesting published results on Glu transporters membrane topology, Glu transporters physiopathological role and Glu transporters medicinal chemistry, highlighting the guidelines for the development of potential neuroprotective agents targeting neuronal high-affinity sodium-dependent glutamate transporters. PMID- 12570796 TI - The role of melanocortin peptides and receptors in regulation of energy balance. AB - Energy balance is a highly regulated, complex process which is modulated by central and peripheral systems. Dysregulation of energy homeostasis can result in metabolic disorders, such as obesity and type II diabetes. Obesity and type II diabetes are two of the most prevalent and challenging clinical conditions in society today. A growing body of evidence has implicated the melanocortin system as an important component in the maintenance of energy balance. alpha-MSH, a 13 amino acid peptide secreted as a product of the pro-opiomelanocortin (POMC) gene in the pituitary is a potent agonist of 4 of the 5 cloned melanocortin receptors (MCR). MC receptors are members of a G-protein-coupled receptor (GPCR) family, which signal through cAMP. Agouti and agouti-related protein (AGRP) are natural antagonists of melanocortin receptors and participate in regulation of skin/fur pigmentation, body weight, and adiposity. Stimulation of MC receptors has pleiotropic effects, which impact the nervous system as well as endocrine and immune functions. One of the most prominent effects of MC receptor stimulation is a dramatic suppression of food intake and body weight, which has led to the hypothesis that the MC receptor system plays a primary role in the maintenance of energy balance. This idea is supported by a large body of pharmacological, molecular and human genetic evidence. The following review summarizes the role of melanocortin receptors in the regulation of food intake and energy homeostasis and highlights the opportunities for MC receptors as drug development targets in treating eating disorders and diabetes. PMID- 12570797 TI - Progress in developing D3 dopamine receptor ligands as potential therapeutic agents for neurological and neuropsychiatric disorders. AB - The dopamine D3 receptor has been the subject of a tremendous amount of research since its discovery in 1990. A previous review of this subject [3] described the advances in molecular biology and neuroanatomical localization of the D3 receptor, with a special emphasis on schizophrenia. In the current review, we attempt to describe recent advances in the biochemistry and pharmacology of the D3 receptor from the molecular to the behavioral level. Evidence linking an alteration in D3 receptor function as playing an important role in the etiology of a variety of CNS disorders, including schizophrenia, Parkinson's Disease, and substance abuse is also provided. Also discussed are the recent developments in attempting to map the ligand-binding domains of the D2 and D3 receptors. A. survey of the literature, including a description of the medicinal chemistry approaches toward developing D3-selective ligands, is also presented in this review. PMID- 12570798 TI - The discovery of epidepride and its analogs as high-affinity radioligands for imaging extrastriatal dopamine D(2) receptors in human brain. AB - [(123)I]Epidepride, [(18)F]fallypride, and [(76)Br]isoremoxipride (FLB-457) and their corresponding [(11)C]labeled derivatives belong to a class of high-affinity radioligands for SPECT or PET imaging of dopamine D(2) receptors in the human brain. In contrast to previously used imaging agents, these ligands are capable of identifying extrastriatal dopamine D(2) receptors. The design of these substituted benzamides derive its origin from the atypical antipsychotic agent, remoxipride. Starting in the late 1970's, halogenated analogs of (S)-sulpiride were evaluated in binding assays and behavioral studies, leading to the discovery of remoxipride. Remoxipride was 10 times weaker than sulpiride in vitro but 50 times more potent in vivo. Search for a putative active metabolite of remoxipride led to the discovery of raclopride and eticlopride, the former becoming a useful radioligand as tritium or carbon-11 labeled form for receptor binding and PET studies, respectively. In the US, the mono-iodine analog of raclopride, [(123)I]iodobenzamide (IBZM), was found to have moderate putamen-to-cerebellum ratio in rat and human brain. Continued search for metabolites of remoxipride led to the discovery of its 3,6-dihydroxy derivative, NCQ-344, with an extremely potent in vivo activity in the rat. SAR studies of the metabolites of remoxipride led to the discovery of the 3-methoxy isomer, isoremoxipride (FLB-457) and its corresponding 6-hydroxy analog, FLB-463, both having affinities for the dopamine D(2) receptor in the 20-30 pM range. Later, the 5-[(123)I]iodo analog of FLB-463, [(123)I]ioxipride ([(123)I]NCQ-298), became a potential SPECT imaging agent. In the mean time, the deshydroxy analog of IBZM, [(125)I]iodopride, showed binding potential in the rat similar to [(125)I]IBZM. Epidepride was designed by combining the structure of isoremoxipride with that of iodopride. In 1988, epidepride was independently prepared and radiolabeled in three separate laboratories in Stockholm, Berkeley, and Nashville. Evaluation of seven [(125)I]iodine substituted analogs of raclopride, including IBZM, revealed the unusual high striatum-to-cerebellum ratio of 234 of [(125)I]epidepride in the rat. Subsequent SPECT images with [(123)I]epidepride demonstrated its ability to identify extrastriatal dopamine D(2) receptors in the human brain. Exploration of the structure of epidepride confirmed its exceptional properties, to be exceeded only by its N-allyl homolog, [(125)I]nalepride. The design by others of a series of potent 5-(3-[(18)F]fluoropropyl) substituted analogs of epidepride for PET imaging, lead to the discovery of [(18)F]fallypride. By elucidating the role of lipophilicity in the substituted benzamides, the excellent imaging characteristics of [(11)C]/[(123)I]epidepride, [(11)C]/[(76)Br]isoremoxipride and [(18)F]fallypride, could not only be explained but predicted with remarkable accuracy. By using the inverse product of the receptor affinity (K(D)), and the apparent partition constant of the radioligand (P((7.4))), estimates of maximal binding potential of any radioligand for imaging of any neurotransmitter receptor or transporter site seem possible. PMID- 12570799 TI - Regulation of angiogenesis by Th1- and Th2-type cytokines. AB - Angiogenesis is a complex process, where several cell types and mediators interact to establish a specific microenvironment suitable for the formation of new capillaries from pre-existing vessels. Such biological processes occur in several physiological conditions, such as embryo development and wound healing, as well as in pathological conditions, including tumours and diabetic retinopathy. T lymphocytes, neutrophils and monocytes fully participate in the angiogenic process by secreting cytokines that may control endothelial cell (EC) proliferation, their survival and apoptosis, as well as their migration and activation. Angiogenesis is the result of a net balance between the activities exerted by positive and negative regulators. This balance is conceptually very similar to that of the Th1/Th2 cells that modulate an appropriate and specific immune response. Th1 or Th2 cytokines may control angiogenesis directly, by acting on cell growth and differentiation, indirectly by inducing the release of other cytokines in the microenvironment, and by modulating the expression of specific receptors, involved in the control of angiogenic processes, such as EC proliferation and migration. In this review we will mainly discuss the role of Th1- and Th2-type cytokines in the angiogenic process, emphasizing the complexity of the cytokine and leukocyte/EC network, and highlighting the care that needs to be taken when designing new therapeutic interventions involving Th1 and Th2 cytokines. PMID- 12570800 TI - Role of nitric oxide in the modulation of angiogenesis. AB - Angiogenesis, the development of new capillaries form pre-existing vessels, requires the coordinate activation of endothelial cells, which migrate and proliferate in response to growth factors to form functional vessels. Therapeutic angiogenesis is proposed to restore tissue integrity and function following damage and ischemia, while strategies aimed to block or suppress the neovascular growth are designed as adjuvant therapies for cancer treatment. Different experimental and clinical observations support the existence of a molecular/biochemical link between vasodilation, nitric oxide (NO) production and angiogenesis. NO significantly contributes to the prosurvival/proangiogenic program of capillary endothelium by triggering cell growth and differentiation via endothelial-constitutive NO synthase (ecNOS) activation, and cyclic GMP (cGMP) dependent gene transcription. Re-establishment of a balanced NO production in the cardiovascular system results in a reduction of cell damage during inflammatory and vascular diseases. Elevation of NOS activity in correlation with angiogenesis and tumor growth and aggressiveness has been extensively reported in experimental and human tumors. On these bases, the nitric oxide pathway appears to be a promising target for the development of pro- and anti-angiogenic therapeutic strategies. In particular, the use of NOS inhibitors or NO scavengers seems appropriate to reduce edema, block angiogenesis and facilitate antitumor drug delivery. PMID- 12570801 TI - Protein kinases and the hypoxia-inducible factor-1, two switches in angiogenesis. AB - In the last few decades it has become clear that detailed understanding of the mechanisms of angiogenesis, a process leading to growth of new blood vessels, should lead to improved treatment of diseases such as ischemic disorders and cancer where neovascularization is impaired or activated, respectively. In this review, we will outline some of our recent findings concerning the regulation of the vascular endothelial growth factor (VEGF), a key player in angiogenesis and one of its transcription factors, the hypoxia-inducible factor-1 (HIF-1) a master gene product driving adaptation to hypoxia. We will discuss the observation that growth factors and oncogenic transformation via the mitogen-activated protein kinases p42/p44 MAPKs not only activate the VEGF promoter through the Sp1/AP-2 transcriptional factor complex but also phosphorylate HIF-1alpha leading in turn to enhance HIF-1 dependent transcriptional activation of VEGF. The stress activated protein kinases (SAPK) also contribute to angiogenesis by stabilizing VEGF mRNA. Finally, we will present recent advances into oxygen-sensing, in particular the HIF-hydroxylases that govern HIF-1alpha instability (PHD2) or inactivation (FIH-1). The revelation of these oxygen sensors has provided pharmacologists with new molecular targets for the development of novel therapies to control angiogenesis either positively or negatively. PMID- 12570802 TI - Calcium as a molecular target in angiogenesis. AB - The plethora of cellular pathways and events involved in angiogenesis are a prime example of the widespread role of calcium ion flux in biological functions. Indeed, calcium is a main point of intersection for many distinct molecular signaling pathways that promote and modulate angiogenesis. Here, we illustrate some of the important aspects of calcium induction, function, downstream effects, and resulting cellular changes that ensue. We describe some of the main mechanisms of calcium regulation in cells as well as intracellular and cross membrane flux, highlighting key players that are known to facilitate these events. We review some of the major signaling pathways that tie into angiogenesis, and also describe how cellular phenotypic changes that occur during angiogenesis require processes rich in calcium ion stimulation of gradient shifts. Lastly, we hypothesize on current thinking of the role of calcium as a whole in angiogenic cellular function and propose new insight into calcium as a universal effector molecule and a prime target for therapeutic intervention. PMID- 12570803 TI - Heparin derivatives as angiogenesis inhibitors. AB - Angiogenesis is the process of generating new capillary blood vessels. Uncontrolled endothelial cell proliferation is observed in tumor neovascularization and in angioproliferative diseases. Tumors cannot growth as a mass above few mm(3) unless a new blood supply is induced. It derives that the control of the neovascularization process may affect tumor growth and may represent a novel approach to tumor therapy. Angiogenesis is controlled by a balance between proangiogenic and antiangiogenic factors. The angiogenic switch represents the net result of the activity of angiogenic stimulators and inhibitors, suggesting that counteracting even a single major angiogenic factor could shift the balance towards inhibition. Heparan sulfate proteoglycans are involved in the modulation of the neovascularization that takes place in different physiological and pathological conditions. This modulation occurs through the interaction with angiogenic growth factors or with negative regulators of angiogenesis. Thus, the study of the biochemical bases of this interaction may help to design glycosaminoglycan analogs endowed with angiostatic properties. The purpose of this review is to provide an overview of the structure/function of heparan sulfate proteoglycans in endothelial cells and to summarize the angiostatic properties of synthetic heparin-like compounds, chemically modified heparins, and biotechnological heparins. PMID- 12570804 TI - Small-molecule inhibitors of fibroblast growth factor receptor (FGFR) tyrosine kinases (TK). AB - Fibroblast growth factor receptors (FGFR) are members of a family of polypeptides synthesized by a variety of cell types during the processes of embryonic development and in adult tissues. FGFR have been detected in normal and malignant cells and are involved in biological events that include mitogenic and angiogenic activity with a consequent crucial role in cell differentiation and development. To activate signal transduction pathways, FGFR are coupled to fibroblast growth factors (FGF) and heparan sulfate (HS) proteoglycans to form a biologically fundamental ternary complex. Based on these considerations, a variety of inhibitors able to block the signaling cascade through a direct interaction with FGFR have been designed and investigated for their biological properties related to antiangiogenesis and antitumor activity. The purpose of this review is to focus on synthetic chemical approaches aimed at blocking tyrosine kinase (TK) receptors, members of the FGFR family. In particular, a literature survey aimed at summarizing on the structural properties that a compound should possess to show affinity toward FGFR is presented, and structure-activity relationships (SAR) on FGFR inhibitors are delined. PMID- 12570805 TI - Thrombospondins as anti-angiogenic therapeutic agents. AB - Thrombospondin-1 (TSP-1) was one of the first endogenous inhibitors of angiogenesis to be discovered. This large multimodular protein of around 600 kDa inhibits endothelial cell proliferation, migration and morphogenic organization into capillary tubes. TSP-2 shares homology with TSP-1 in primary sequence, structural organization and angiostatic properties. TSP-1-null and TSP-2-null mice display increased tissue vascularity and enhanced sensitivity to carcinogenesis. Conversely, overexpression of TSP-1 or TSP-2 in cancer cells results in reduced tumor vascularization and tumor growth. In this review, we focus on the preclinical data obtained in experimental anti-tumorigenic assays using either TSP-1, TSP-2 or shorter peptides derived from the type 1 repeats of these molecules. In contrast with the full length thrombospondin molecules, which present a poor bioavailability and are highly susceptible to proteolytic degradation, TSP-derived angiostatic peptides appear as potent and promising therapeutic agents in anti-angiogenic therapy. PMID- 12570806 TI - Human tissue kallikrein: a new bullet for the treatment of ischemia. AB - Recently, therapeutic angiogenesis has been proposed as an alternative for the treatment of ischemic diseases unresponsive to conventional therapy. This strategy is based on the concept that a supply-side approach with growth factors would overcome the endogenous deficit and result in more robust collateralization. We have developed a strategy based on local delivery of human tissue kallikrein gene for potentiation of microcirculation and rescue of peripheral ischemia. Following successful application in otherwise healthy animals, the approach resulted to be of therapeutic value in rats with endothelial dysfunction caused by arterial hypertension. In addition, human tissue kallikrein prevents or rescues microvascular rarefaction caused by diabetes mellitus. In this model, human tissue kallikrein was able to stimulate vascular growth and contrast apoptosis. The strategy displays interesting pharmacological features because is devoid of obvious side effects and is effective even at low infecting doses. In addition, the neovascularization promoted by human tissue kallikrein is well organized and durable. It is reasonable to anticipate that the new approach will have a great impact in the treatment of cardiovascular ischemic complications. PMID- 12570807 TI - Emerging beta-amyloid therapies for the treatment of Alzheimer's disease. AB - Alzheimer's Disease (AD) is a progressive neurodegenerative disorder marked by loss of memory, cognition, and behavioral stability. AD is defined pathologically by extracellular neuritic plaques comprised of fibrillar deposits of beta-amyloid peptide (Abeta) and neurofibrillary tangles comprised of paired helical filaments of hyperphosphorylated tau. Current therapies for AD, such as cholinesterase inhibitors, treat the symptoms but do not modify the progression of the disease. The etiology of AD is unclear. However, data from familial AD mutations (FAD) strongly support the "amyloid cascade hypothesis" of AD, i.e. that neurodegeneration in AD is initiated by the formation of neurotoxic beta-amyloid (Abeta) aggregates; all FAD mutations increase levels of Abeta peptide or density of Abeta deposits. The likely link between Abeta aggregation and AD pathology emphasizes the need for a better understanding of the mechanisms of Abeta production. This review summarizes current therapeutic strategies directed at lowering Abeta levels and decreasing levels of toxic Abeta aggregates through (1) inhibition of the processing of amyloid precursor protein (APP) to Abeta peptide, (2) inhibition, reversal or clearance of Abeta aggregation, (3) cholesterol reduction and (4) Abeta immunization. PMID- 12570808 TI - Peptidases, proteases and amyloid beta-peptide catabolism. AB - The formation of senile plaques containing amyloid beta peptides (Abeta peptides) as a major constituent plays a significant role in development of Alzheimer's disease. The concentration of Abeta peptides in the brain is determined by a combination of their rate of synthesis and their rate of clearance. Considerable effort has been expended in producing inhibitors of the beta and gamma secretases involved in the synthesis of the Abeta peptides. More recently interest in the mechanism of clearance of the Abeta peptides has emerged, as promoting Abeta peptide clearance represents an alternative therapeutic approach. It now appears that cleavage of Abeta peptide by peptidases and proteases represents the major mechanism of clearance. This review describes those peptidases and proteases implicated in Abeta peptide clearance, the evidence that these enzymes function in vivo, and how they may represent new therapeutic targets. PMID- 12570809 TI - Drug-targeting strategies for prostate cancer. AB - Prostate cancer is the most frequently diagnosed cancer in North American men and accounts for 10% of cancer-related deaths in men. Despite advances in early detection and aggressive treatment of early disease, the overall mortality rate has not appear to have fallen, indicating that the current therapies are not beneficial for life expectancy and new strategies are required. Prostate cancer is a dynamic evolving process that develops in distinct steps, with each step liable to additional genetic hits that change the cancer cell phenotype and alter the patterns of gene expression. The molecular events in prostate cancer are beginning to be understood, including altered expression of tumor suppressor genes, pro- and anti-apoptotic genes, and oncogenes associated with the progression of the disease; and specific genes that are expressed predominantly or exclusively in prostate cells, prostate cancer cells, and prostate metastasis cells. These latter genes on the level of DNA, RNA and protein products are the targets of several new approaches to prostate cancer therapy and are the focus of this review. PMID- 12570810 TI - PACAP in avians: origin, occurrence, and receptors--pharmacological and functional considerations. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a novel member of the secretin/glucagon/vasoactive intestinal peptide (VIP) superfamily. In vertebrates, including avians, it occurs in two forms: PACAP(38) and PACAP(27). PACAP structure is well conserved during evolution, being identical in mammals, and showing one amino acid dfifference in avians (chick, turkey). PACAP is widely distributed in the central nervous system and peripheral tissues and displays a pleiotropic activity, including functions as a hypophysiotropic hormone, neuromodulator, and neurotrophic factor. PACAP exerts its biological actions through three types of receptors designated PAC(1), VPAC(1) and VPAC(1). This review (1) presents the current knowledge on PACAP origin, distribution and function, (2) compares the avian findings with those found in mammals, and (3) describes receptor-linked mechanisms in avians, including recent data on receptor related signal transduction pathways, with a special emphasis on receptor pharmacology and function. PMID- 12570811 TI - VIP and drug design. AB - The following review outlines the physiological outcome of VIP and VIP gene manipulations. Previously, we reviewed the various VIP receptors associated with biological functions ranging from growth regulation, sexual function, bronchodilation, vasodilation and immune interactions to neurotrophism. VIP-based drug design is discussed below. PMID- 12570813 TI - Neuropeptides as autocrine growth factors in cancer cells. AB - Neuropeptides can function as autocrine growth factors in cancer cells. High levels of bombesin (BB) and neurotensin (NT)-like immunoreactivity are present in small cell lung cancer (SCLC), a neuroendocrine tumor. Vasoactive intestinal peptide (VIP) stimulates and somatostatin (SST) inhibits the release of BB-like peptides from SCLC cells. BB-like peptides bind to BB(2) receptors, which are present on the cell surface. BB-like peptides stimulate the mitogen activated protein kinase (MAPK) cascade leading to increased expression of nuclear oncogenes and growth factors in SCLC cells. Due to the high density of neuropeptide receptors present on the cell surface, SST analogs have been radiolabeled to image neuroendocrine tumors. VIP receptors are present in many epithelial cancers including breast, colon, non-small cell lung cancer (NSCLC), pancreatic and prostate cancers. Due to the high density of VIP receptors on lung cancer cells, radiolabeled VIP agonists may be used to image these tumors. VIP receptor antagonists, such as VIPhybrid, inhibit the growth of cancer cell lines in vitro and in vivo. VIPhybrid and SR48692, a NT receptor antagonist, potentiate the cytotoxicity of chemotherapeutic drugs. These results suggest that neuropeptide receptor antagonists may be useful in the treatment of cancer. PMID- 12570814 TI - Nitric oxide and its role during pregnancy: from ovulation to delivery. AB - Nitric oxide (NO) is a major paracrine mediator and important regulatory agent in various female reproductive processes, such as ovulation, implantation, pregnancy maintenance, labor and delivery. Ovulation: Circulating NO-products are increased during follicle development and decreased right after ovulation. INOS-inhibition results in a 50% reduction of ovulation, an effect completely reversed by an NO. Endometrium/Implantation: NO also regulates endometrial functions such as endometrial receptivity, implantation and menstruation. NO-donors may be useful for promoting fertility, while NO-inhibitors might be used for contraception. Uterine contractility: Throughout gestation myometrial NO-production is upregulated thus contributing to achieve uterine quiescence. Close to term, NO production decreases promoting effective contractions resulting in labor. Clinical trials have demonstrated that NO-donors are effective tocolytics. Cervical ripening: In contrast to the myometrium, NO-production in the cervix is low during gestation and becomes upregulated once pregnancy advances to term. NO donors are effective and safe cervical ripening agents. This finding from animal studies has been confirmed by several clinical trials. Vasoreactivity: In blood vessels, NO is a potent vasodilator and platelet-aggregation-inhibitor. Lack of NO during gestation was related to the development of pregnancy-induced hypertension and preeclampsia. In conclusion, NO-donors and NOS-inhibitors may provide novel, effective, safe, and inexpensive drugs to regulate and steer various functions in female reproductive life. The benefits reach from contraception to preventing possibly lethal pregnancy complications such as preeclampsia. Introducing NO-donors as tocolytics and cervical ripening agents may contribute to a reduction of fetal and maternal perinatal morbidity and mortality. PMID- 12570815 TI - The role of nitric oxide (NO) in control of LHRH release that mediates gonadotropin release and sexual behavior. AB - Nitric oxide (NO) plays a crucial role in reproduction at every level in the organism. In the brain, it activates the release of luteinizing hormone-releasing hormone (LHRH). The axons of the LHRH neurons project to the mating centers in the brain stem and by efferent pathways, evoke the lordosis reflex in female rats. In males, there is activation of NOergic terminals that release NO in the corpora cavernosa penis to induce erection by generation of cyclic guanosine monophosphate (cGMP). NO also activates the release of LHRH which reaches the pituitary and activates the release of gonadotropins by activating neural NO synthase (NOS) in the pituitary gland. Follicle stimulating hormone (FSH)RH selectively releases FSH also by activating NOS. Leptin releases LHRH by activating NOS to release FSH and LH with the same potency as LHRH. These actions are mediated by specific receptors on the gonadotropes for LHRH, FSHRH and leptin. The responsiveness of the pituitary is controlled by gonadal steroids. In the gonad, NO plays an important role inducing ovulation and in causing luteolysis; whereas in the reproductive tract, it relaxes uterine muscle via cGMP and constricts it by prostaglandins. PMID- 12570816 TI - Mouse models and the role of nitric oxide in reproduction. AB - As a group, nitric oxide synthase (NOS) isoforms are localized to a wide variety of tissues. Understanding the role of NOS in reproductive physiology was facilitated by the introduction of genetically engineered mice. Specifically, "knock-out" mice with targeted disruptions in neuronal NOS, endothelial NOS and inducible NOS have been made. These models have been useful in addressing the of role of nitric oxide in areas of reproductive biology that include: hypothalamic pituitary-ovarian axis, mating behavior, maternal blood pressure regulation and fetal development. Despite several promising observations using "knock-out" mouse models, one must exercise caution in interpreting data from individual experiments. Very often the need to draw on two mouse strains to generate the founder NOS deficient line, purity of the strain used for observing phenotypes (presence or absence of backcrossing to achieve a pure strain), and the strain used for comparison purposes (control strains are often not identical in their genetic make-up to the deficient line) make wide spread applicability of results open to criticism. PMID- 12570817 TI - Nitric oxide in oocyte maturation, ovulation, fertilization, cleavage and implantation: a little dab'll do ya. AB - Nitric oxide synthases, the enzymes that generate NO gas, may be involved in reproduction and development of multicellular organisms at many levels and thus provide important targets for design of drugs to intervene in reproductive processes. This review focuses on the role of nitric oxide in key events of reproduction including gamete activation, fertilization, early cell divisions and implantation. A general trend highlighted by the studies reviewed is that NO plays a biphasic role in reproduction. That is, a narrow range of NO concentrations, usually low, will stimulate or enhance these early events in reproduction, but either a lack of NO or too much NO has negative consequences. One of the shortcomings of the field currently is the lack of molecular detail concerning the mechanism of NO action. This has been due in part to lack of technology for effective detection of NO and its molecular targets. A few targets of NO have been indirectly implicated and advances in this area of research will provide substrates for development of drugs to control reproductive function. Work from both invertebrate and vertebrate model systems is presented and implications for control of reproductive physiology discussed. Ubiquity of NO signaling in animals may mean that effective control of reproduction must target mediators of NO action and not NOS enzymes themselves. PMID- 12570818 TI - Nitric oxide regulation of permeability in human cervical and vaginal epithelial cells and in human endothelial cells. AB - This paper reviews new models of nitric-oxide (NO)-related regulation of transepithelial and transendothelial permeability in normal tissues, with emphasis on physiological relevance of the data. Novel data obtained in cultured human epithelial and endothelial cells indicate that NO can originate from different intracellular sources, and can be selectively regulated and employed to activate various intracellular mechanisms that will affect the permeability via different mechanisms. These models may be important for understanding NO regulation of permeability in vivo. PMID- 12570819 TI - Nitric oxide is a signaling molecule in spermatozoa. AB - Low and controlled concentrations of nitric oxide play an important role in sperm physiology. Nitric oxide is produced by spermatozoa and acts as an intracellular signaling molecule in the processes of capacitation and acrosome reaction. It has been documented that during capacitation, nitric oxide interacts with the cAMP protein kinase A pathway and also is involved in tyrosine nitration of sperm proteins. On the other hand, during the acrosome reaction, two different pathways have been postulated for nitric oxide to exert its effects. During the progesterone-induced acrosome reaction, nitric oxide stimulates a heme-containing enzyme, named cyclooxygenase with a subsequent increase in prostaglandin E(2). Furthermore, the acrosome reaction inducing effect of NO-releasing compounds occurs via an increase in cGMP levels and protein kinase G activation. Taken together, these data support a role for nitric oxide in sperm function. This review focuses on providing new evidence for the physiological role of nitric oxide (NO) on sperm function. We will first present a brief description on nitric oxide chemistry and on the events leading to sperm fertilizing ability followed by the observations obtained on the participation of NO on fertilization. PMID- 12570820 TI - Anti-TNF therapy for Crohn's disease. AB - Although the cause of Crohn's disease remains unknown considerable progress has been made in recent years to unravel the pathogenesis of the inflammatory processes seen in chronic idiopathic inflammatory bowel diseases (IBD). Th-1 lymphocytes seem to orchestrate the inflammation through the production of pro inflammatory cytokines such as IFN-gamma, IL-1beta and tumor necrosis factor (TNF). After isolation and characterization of TNF and its two receptors (p55 and p75) detailed regulatory processes for transcription, secretion and post receptor actions of TNF are now rapidly being discovered. Genetically engineered monoclonal antibodies, specifically directed against TNF are only the first drugs acting against TNF, available for clinical use now in the treatment of Crohn's disease. A single IV injection of these antibodies produces very dramatic clinical, endoscopic and histological responses in a majority of refractory patients. More data on long term safety and the exact role in combination with standard therapies are being awaited. In the mean time, these drugs should be reserved for patients not responding to standard antiinflammatory therapy. The exciting "TNF story" very nicely illustrates how the benchmark of basic immunological research now provides us with very potent and rationally designed drugs. Expected and unexpected safety toxicity data should caution clinicians to a certain extent against too liberal use of these agents interfering with very basic physiological events. PMID- 12570821 TI - Modification of intestinal flora in the treatment of inflammatory bowel disease. AB - Because the intestinal microflora play an important role in the development of inflammatory bowel disease (IBD), there is currently some interest in the manipulation of the composition of the microflora towards a potentially more remedial community. This review summarizes the clinical and experimental efficacy of the manipulation of microflora by the use of prebiotics, probiotics, synbiotics, and antibiotics in IBD. Prebiotics, defined as nondigestible food ingredients that beneficially affect the host by selectively stimulating the growth or activity of one or a limited number of bacterial species already resident in the colon, can modulate the colonic microbiota by increasing the number of specific bacteria and thus changing the composition of the microbiota. Prebiotics for IBD include lactosucrose, oligofructose, inulin, bran, psyllium, and germinated barley foodstuff (GBF). GBF, which mainly consists of dietary fiber and glutamine-rich protein, is a prebiotic foodstuff for ulcerative colitis. GBF has shown to be converted into a preferential nutrient for colonocytes through Eubacterium and Bifidobacterium and also inactivate nuclear factor kappa B (NFkB). Moreover, it exhibits a potent water-holding capacity and bile-acid binding capacity. Probiotics, which are microbial food supplements that beneficially affect the host by improving the intestinal microbial balance, have been used to change the composition of colonic microbiota. The approaches for IBD include VSL#3, Nissle1917, Clostridium butyricum and Bifidobacterium-fermented milk. Use of Lactococci secreting IL-10 provides excellent results. The combination of prebiotics and probiotics in a synbiotic has not been studied in IBD but is promising. The use of antibiotics continues to be of interest. Although these strategies hold great promise and appear to be useful in some settings, more clinical study is needed to firmly establish the relevance of these therapies. PMID- 12570822 TI - Anti-interleukin-6 therapy for Crohn's disease. AB - Proinflammatory cytokines have been demonstrated to play a crucial role in the pathogenesis and physiopathology of various chronic inflammatory conditions including Crohn's disease (CD). Among these cytokines, interleukin-6 (IL-6) must be especially important because increased serum concentrations of acute phase proteins, reduced level of serum albumin, and remarkable thrombocytosis are all well-explained by the increased level of IL-6. Moreover, IL-6 is capable of stimulating even IL-6 receptor (IL-6R) negative cells such as vascular endothelial cells when complexed to soluble form of IL-6R (sIL-6R), and serum level of IL-6 as well as sIL-6R has been demonstrated to increase during inflammation. To investigate the therapeutic potential of IL-6 signaling blockade for CD, anti-IL-6R monoclonal antibody (mAb) was introduced to various murine models of colitis. Anti-IL-6R mAb successfully prevented wasting disease and the development of macroscopic and histological lesions. It suppressed the accumulation of ICAM-1 positive and Mac-1 positive cells in the lamina propria (LP) and the expression of ICAM-1 and VCAM-1 by vascular endothelial cells. Expansion of colonic and splenic CD4(+) T cells was reduced as well as the colonic expression of tumor necrosis factor alpha (TNF-alpha), IL-1beta, and interferon gamma (IFN-gamma) mRNA without affecting the production of transforming growth factor beta (TGF-beta), IL-10, and IL-4 mRNA. The treatment also suppressed established colitis by inducing LP T cell apoptosis. These results strongly suggest that specific targeting of IL-6/sIL-6R pathway will be a promising new approach for the treatment of CD, and the clinical trial of humanized anti-IL-6R mAb is now under way. PMID- 12570823 TI - Multicenter randomized controlled trial for the treatment of ulcerative colitis with a leukocytapheresis column. AB - The administration of steroids is not always effective for the treatment of ulcerative colitis (UC). Their long-term use often causes adverse effects which sometimes result in their stoppage and acute exacerbation. Therefore, an alternative treatment is necessary in order to decrease steroid dosage and avoid the clinical problems associated with steroids. Methods The effectiveness and adverse effects of a leukocytapheresis (LCAP) were investigated in a controlled multicenter trial with randomized assignment of 76 active-stage UC patients in two groups. In the LCAP group (39 patients), LCAP weekly for 5 weeks as an intensive therapy was added to the on-going drug therapy, while steroids were maintained but not increased, and then LCAP was gradually reduced to once every 4 weeks as a maintenance therapy. In the high dose prednisolone (h-PSL) group (37 patients), PSL was added or increased 30 approximately 40 mg/day for moderately severe and 60 approximately 80 mg/day for severe patients and then gradually tapered. Findings The LCAP group showed a significantly higher effectiveness (74% vs. 38%; p=0.005) and lower incidence of adverse effects (24% vs. 68%; p<0.001). The patients were able to continue the trial for a longer period in the LCAP group than the h-PSL group (p=0.012). Clinical activity and endoscopic indexes showed the LCAP group had better improvements than the h-PSL group. Interpretation The results of the trial show that LCAP permits a reduction in total PSL dosage and is more effective and safer than high-dose PSL administration for intensive therapy, and LCAP may maintain remission longer than PSL. PMID- 12570824 TI - Enteral and parenteral nutrition therapy for Crohn's disease. AB - Even with the development of new therapeutic agents, such as infliximab, enteral nutrition (EN) and parenteral nutrition (PN) therapies remain important for the treatment of Crohn's disease because Crohn's patients often require nutritional support. Furthermore, nutritional therapies can be used in the control of disease activity. Elemental diets, which are mainly used in EN therapy, consist of a refined amino acid mixture, glucose or maltodextrins and minimal essential fatty acids. EN therapy can reduce mucosal inflammation by the elimination of dietary antigens, which induce inflammation, and by reductions in fat, which activates inflammation. EN is applied not only as induction therapy, but also as maintenance therapy after remission (home EN). However, the unpalatability of elemental diets, difficulties related to self-intubation and the high cost of EN have limited its application as a primary therapy in western countries. PN is utilized as complete bowel rest supporting nutrition. However, since the therapeutic efficacies of EN and PN are similar, the indications for PN are limited and PN is mainly utilized in patients with bowel obstructions or severe fistulas. PN is also used as home therapy in the treatment of Crohn's patients with short bowel syndrome. However, long-term PN sometimes causes life threatening complications including catheter-induced sepsis, liver failure and lethal mineral deficiencies. We suggest that gastroenterologists should recognize the advantages and limitations of all therapies and choose carefully or combine various therapies in order to maintain the quality of life in individual patients even if in cases where remission can not be achieved. PMID- 12570825 TI - Role of dietary fiber and short-chain fatty acids in the colon. AB - Luminal nutrition is important for maintenance of gastrointestinal mucosal structure and function. In particular, short chain fatty acids (SCFAs), metabolic products of anaerobic bacterial fermentation of dietary fiber and resistant starch, are particularly important as the preferred respiratory fuel of the colonocytes. A variety of biological effects of SCFAs have been reported, and there is now increasing number of experimental works showing new aspects of these molecules. For example, as the mechanisms mediating anti-inflammatory effects of SCFAs, several investigators identified the inhibitory effect of butyrate on proinflammatory cytokine-induced NF-kappaB activation. Various inflammatory responses are now discussed with the central role of NF-kappaB activation, and thus the inhibition of NF-kappaB activation represents the efficacy of dietary fiber and SCFAs in the treatment with inflammatory bowel disease. Furthermore, recent advance in molecular technology has identified mechanisms mediating anti tumor effects of SCFAs. SCFAs modulate expression of cell cycle-regulating proteins and induce apoptosis in colon cancer cells. SCFAs increase the susceptibility of colon cancer cells to complement-mediated cell injury. In this review, new aspects of functions of SCFAs are focused and summarized. PMID- 12570826 TI - Natural killer T cells as targets for therapeutic intervention in autoimmune diseases. AB - Natural killer T (NKT) cells are a subset of lymphocytes that express receptors characteristic of conventional T cells together with receptors typically found on natural killer cells. A key feature of NKT cells is the expression of a semi invariant T cell receptor that is specific for glycolipid antigens presented by the unusual major histocompatibility complex class I-like molecule CD1d. While their precise immunological functions remain unknown, NKT cells have been implicated in the regulation of adaptive immune responses, including those directed against autoantigens. These findings raise the possibility that specific stimulation of NKT cells may be exploited for therapeutic purposes. A number of laboratories have tested this hypothesis, utilizing the sea sponge-derived agent alpha-galactosylceramide (alpha-GalCer), a specific agonist of NKT cells. Administration of alpha-GalCer to mice results in potent activation of NKT cells, rapid and robust cytokine production, and activation of a variety of cells of the innate and adaptive immune systems. Most notably, repeated administration of alpha-GalCer to mice favors the generation of conventional T lymphocytes producing T helper (Th) type 2 cytokines such as IL-4 and IL-10. These findings suggest that alpha-GalCer can modulate inflammatory conditions that are mediated by pathogenic Th1 cells. Indeed, recent studies have demonstrated that alpha GalCer prevents the development of Type 1 diabetes in non-obese diabetic mice and central nervous system inflammation in mouse models of multiple sclerosis. Collectively, these studies provide a solid foundation for the development of NKT cell ligands as pharmacological agents for treatment of autoimmune diseases. PMID- 12570827 TI - Dendritic cell homeostasis in the regulation of self-reactivity. AB - Dendritic cells (DC) are known for their remarkable ability to induce specific T cell responses. However, the existing views on the role of DC in maintaining tolerance to self-antigens and induction of autoimmunity are somewhat controversial especially when the basic physiology of DC migration, function and homeostasis is considered. This review attempts to provide a comprehensive overview on these topics with particular emphasis on DC homeostasis and presents implications for the generation of pathological autoimmune T cell responses. Furthermore, we advocate the need for a conceptual characterization of the immune system operating in vivo. With particular focus on the contribution of DC, we suggest that a 'spatiotemporal' view of the rules for T cell responses (antigen dose and availability, duration and mode of antigen presentation) permits a better understanding of the relevant factors contributing to the pathogenesis of autoimmune diseases. PMID- 12570828 TI - Modulation of T cell immunity by TCR/pMHC dwell time and activating/inhibitory receptor pairs on the antigen-presenting cell. AB - The molecular interactions occurring at the interface between the antigen presenting cell (APC) and the T lymphocyte play an important role in the immune surveillance against infectious agents and tumors, as well as in autoimmunity and transplant rejection. The significance of the APC-T cell interaction in immunity is underscored by the observation that deficiencies in the function of either one of these two cell types cause extreme susceptibility to infections and tumor growth. Furthermore, a disregulated APC-T cell interaction can initiate autoimmunity. Thus, antigen recognition by T cells must be tightly regulated in order to ensure protection against pathogens and tumors, avoiding activation of self-reactive T cells. Efficient T cell activation requires two simultaneous signals provided by the APC: Antigen (or signal 1) and co-stimulation (or signal 2). The specificity of antigen recognition by T cells (signal 1) is controlled exclusively by the T cell receptor (TCR), an extremely diverse heterodimeric protein composed of disulfide-bonded alpha and beta chains. While it is clear that the TCR recognizes antigens as small peptides bound to molecules of the Major Histocompatibility Complex (MHC), the molecular explanation for the specificity of antigen recognition by the betaalphaTCR is just beginning to be elucidated. In this review are described some of the advances made in the understanding of the molecular interactions that define the antigen-specificity of the TCR, and the current models for T cell activation by antigen on APCs are discussed. PMID- 12570829 TI - Regulation of self-reactive T cells by human immunoglobulins--implications for multiple sclerosis therapy. AB - The intravenous administration of high doses of immunoglobulins pooled from the plasma of healthy donors (IVIg therapy) has beneficial effects in patients with a variety of autoimmune disorders. These clinical observations indicate that IVIg have potent antiinflammatory characteristics, and identification of the precise mode of action may open up perspectives for future therapeutic strategies. In certain tissue-specific autoimmune disorders like multiple sclerosis (MS), self reactive T cells recognizing autoantigens play a significant role for disease pathogenesis, as these cells are able to initiate, maintain, and propagate the harmful immune attack in experimental animal models of disease. These findings render self-reactive T cells an important therapeutic target for autoimmune diseases. Here, we review the effects of IVIg on the homeostasis of T cells and discuss the possible therapeutic implications for multiple sclerosis. As supported by several experimental studies, IVIg regulate crucial steps of T cell mediated immune responses. These effects involve the modulation of activation, proliferation, differentiation, apoptosis, and effector mechanisms of T cells. The pattern of IVIg-T cell interactions is complex, as IVIg may directly bind to regulatory structures on T cells, or modulate T cell functions indirectly via soluble or cellular components of the immune system. PMID- 12570830 TI - Macrophages and the regulation of self-reactive T cells. AB - Macrophages are professional scavengers of apoptotic and necrotic cells, and hence constantly take up self antigens. Paradoxically, macrophages are also professional antigen-presenting cells, which would seem to invite autoimmune disorders. Moreover, macrophages are effector cells in the tissue-destruction phase of autoimmune disorders, where they encounter additional self antigens in the stimulatory context of chronic inflammation. This review examines the array of immunosuppressive mechanisms which may help macrophages suppress unwanted T cell responses, and considers the consequences of a breakdown in these negative regulatory systems in autoimmunity. PMID- 12570831 TI - Autoimmune lymphoproliferative syndrome (ALPS). AB - In patients with ALPS, defective homeostasis of lymphocytes is reflected in abnormal accumulation of lymphocytes, leading to lymphadenopathy, (hepato)splenomegaly and hypersplenism, autoimmunity due to a failure to remove autoreactive lymphocytes, and inappropriate survival of lymphocytes associated with an increased occurrence of lymphoma. Several of the laboratory findings are unique for ALPS and reflect defective Fas-mediated apoptosis and abnormal immune regulation. Much has been learned about the molecular mechanisms that underlie defective Fas-mediated apoptosis and the complex relationship between genotype, phenotype and disease penetrance. Family studies strongly suggest the contribution of one or more additional factors to the pathogenesis of ALPS. This may pertain to defective immunoregulation by an altered IL-2/IL-2 receptor system, reflected in the specific loss of CD4+/CD25+ T cells, and/or by the highly increased IL-10 levels, but other factors may equally be involved. Treatment strategies remain mostly targeted at the disease manifestations, but more specific therapies directed at the primary pathogenic defects themselves might become possible in the future. Continued efforts directed at both careful clinical follow-up and basic scientific investigation are needed to increase our understanding of the incidence, natural history, and pathogenesis of ALPS. In return, this may prove of benefit for the understanding of autoimmune disease in general. PMID- 12570832 TI - Treatment of refractory autoimmune diseases with ablative immunotherapy using monoclonal antibodies and/or high dose chemotherapy with hematopoietic stem cell support. AB - Immunological manipulations are the basis for modern treatments of autoimmune diseases (AID). Targeted immune suppression with lymphopenic based chemotherapy, and monoclonal anti B or T lymphocytic antibodies, are integral part of the conditioning for stem cell transplantation (SCT). Immune manipulation by Cyclophosphamide (Cy), ATG, Campath and recently rituximab (RI), with or without stem cell support are the basis for emerging therapeutic modalities aiming to eradicate the autoreactive clone in various autoimmune disorders. Couple of hundreds of SCTs have been recently performed in various autoimmune disorders, mainly multiple sclerosis (MS), progressive systemic sclerosis (PSS), systemic lupus erythematosis (SLE) and rheumatoid arthritis (RA). Preliminary results are encouraging. Better selection of patients and earlier treatment, before irreversible organ failure develops will probably improve results. Current ongoing multicenter studies are evaluating the role of SCT in MS, RA, SLE, and PSS. PMID- 12570833 TI - Peptide derivatives as agonists or antagonists of formylpeptide receptors: analysis of their effects on neutrophils. AB - The effects of peptide derivatives as agonists or antagonists of formylpeptide receptors are described, taking into account the related cellular responses by neutrophils. These effects are related to the structure of peptide derivatives, some of which are potent anti HIV-1 agents. Finally, formylpeptide receptor models are depicted. PMID- 12570834 TI - Biological role of formaldehyde, and cycles related to methylation, demethylation, and formaldehyde production. AB - An overview is given on the analysis, formation, role and occurrence of formaldehyde in living organisms. Various methods have been used for the determination of formaldehyde in tissues and body fluids. Gas chromatography, thin-layer chromatography and HPLC were employed for the analysis of formaldehyde, mainly after derivatization. The formaldehyde level of human blood and urine was found at the low ppm level. The formaldehyde level could be increased upto several ten micro g/mL(-1) following special dietary supply. Biochemical pathway of both the formaldehyde production and demethylation/methylation processes is generally connected to the methionine - homocysteine cycles. Another important way of demethylation generated formaldehyde production is given by microsomal cytochrome P-450 dependent oxidation of xenobiotics, such as various drugs prescribed by doctors. Semicarbazide sensitive amine oxidase also produces formaldehyde. Increased level of formaldehyde may be the indication of either patho-physiological processes, or environmental contamination, or malnutrition. The formaldehyde-related methylation and demethylation procedures are also detailed. DNA methylation may have an important role in the pathogenesis of certain diseases. PMID- 12570835 TI - Vitamin D receptor as a drug discovery target. AB - 1alpha, 25-dihydroxyvitamin D3 [1,25 (OH)(2)D(3)], the active metabolite of vitamin D3, is known for the maintenance of normal skeleton architecture and mineral homeostasis. Apart form these traditional calcemic actions, 1,25 (OH)(3)D(1) and its synthetic analogs are increasingly recognized for their potent anti-proliferative, prodifferentiative and immunomodulatory activities. The calcemic and non-calcemic actions of 1,25 (OH)(2)D(3) and its synthetic analogs are mediated through vitamin D receptor (VDR), which belongs to the superfamily of steroid/thyroid hormone nuclear receptors. Physiological and pharmacological actions of 1,25 (OH)(2)D(3) in various systems, along with the detection of VDR in target cells, have indicated potential applications of VDR ligands in inflammation, dermatological indications, osteoporosis, cancers and autoimmune diseases. VDR ligands have shown therapeutic potential in limited clinical trials as well as in animal models of these diseases. As a result, a VDR ligand, calcipotriol is in clinic for psoriasis and another, OCT, [2-oxa-1,25 (OH)(2)D(3)] is being developed as a topical agent for the same indication. Further, 1alpha,-hydroxyvitamin D3 (alphacalcidol), a prodrug of 1,25 (OH)(2)D(3) is in clinic and a synthetic VDR ligand, ED-71, is under consideration for approval in Japan for the treatment of osteoporosis. Interestingly, VDR ligands have shown not only preventive but also potent therapeutic anabolic activities in animal models of osteoporosis. However, the wide spread use of VDR ligands in above-mentioned indications is hampered by their major side effect, namely hypercalcemia. In view of this associated toxicity, synthetic VDR ligands with reduced calcemic potential have been synthesized with the ultimate aim of improving their therapeutic efficacy. This review presents recent advances in VDR biology, novel VDR ligands and therapeutic applications of VDR ligands. PMID- 12570836 TI - New aspects of cyclosporin a mode of action: from gene silencing to gene up regulation. AB - Cyclosporin A (CSA) has transformed clinical transplantation, both in term of success and of quality-of-life of the patient. Studies aimed to unfold the site of CSA action have shown that this molecule binds to cytosolic proteins of the cyclophilin family. CSA:cyclophilin complexes have a high affinity for calcineurin, a key enzyme in T-cell activation. By blocking the calcineurin activity, CSA prevents the induction of genes encoding for cytokines and their receptors. Thus, humoral and cellular immune responses are abolished, this resulting in the successful graft acceptance. Disappointingly, CSA and the other molecules as FK506, sharing the capacity to inhibit calcineurin, should be administered for all patient life, as tolerance to alloantigens is not achieved by these molecules. The long term utilization of this class of immunosuppressors increases the incidence of different tumors. The finding that CSA does not interfere with various biochemical pathways has prompted different groups to analyze a possible effect of CSA on molecules that might be involved in different functions of the immune response and/or in tumorogenesis. A new picture of CSA mode of action is emerging in which the immunosuppressor prevents the transcription of a group of genes, concomitantly inducing the transcription of another set. Here, we review the data and discuss the consequences of these new findings in term of T-cell activation mechanisms. PMID- 12570837 TI - The 1,4-dihydropyridine nucleus: a pharmacophoric template part 1. Actions at ion channels. AB - The 1,4-dihydropyridine nifedipine is a prototypical example of the group of calcium channel blockers that also includes a number of second and third generation agents. These drugs enjoy substantial therapeutic prominence for their cardiovascular actions, including hypertension and angina. These actions are exerted at a specific member of the voltage-gated calcium channel family -the L type channel. However, it is increasingly clear that the 1,4-dihydropyridine structure is a pharmacophoric template or "privileged structure" that, when appropriately substituted, can exert potent and selective actions at a diverse set of membrane receptors, including ion channels, G protein-coupled receptors and enzymes. This review will summarize the actions of 1,4-dihydropyridines at these receptors and advance the case that the 4-phenyl-1,4-dihydropyridine structure is a particularly versatile drug template. Part I of the review will summarize actions at ion channels and part II will summarize actions at other receptor systems. PMID- 12570838 TI - Steroid 5alpha-reductase inhibitors. AB - The objective of this study is to synthesize new steroidal compounds based on the progesterone skeleton with a high inhibitory activity for the enzyme 5alpha reductase. Presently similar compounds are being used for the treatment of androgen dependent diseases such as: hirsutism, androgenic alopecia, bening prostatic hyperplasia and prostate cancer. Dihydrotestosterone 2 (Fig. (1)), a 5alpha-reduced metabolite of testosterone 1 has been implicated as a causative factor in the progression of these diseases, largely through the clinical evaluation of males who are genetically deficient of steroid 5alpha-reductase enzyme. As a result of this study, the inhibition of this enzyme has become a pharmacological strategy for the design and synthesis of new antiandrogenic drugs. The advent of finasteride 8 (Fig. (4)) a 5alpha-reductase inhibitor has grately alleviated the symptoms associated with benign prostatic hyperplasia. In our laboratory we recently synthesized several new 16beta-methyl-pregnadiene-3,20 diones derivatives 27 (Fig.(6)), 38-42 (Fig. (11)), 16beta-phenyl-pregnadiene 3,17a-dione derivatives 32-33 (Fig. (7)), 16beta-phenyl-pregnatriene-3,17a diones, 30, 31 (Fig. (7)) and 16beta-methyl-pregnatriene-3,20-diones 43-46 (Fig. (11)). These compounds were evaluated as 5alpha-reductase inhibitors in the following biological models: Penicillium crustosum broths, the flank organs of gonadectomized male hamsters, the incorporation of radiolabeled sodium acetate into lipids, the effect of the new steroids on the reduction of the weight of the seminal vesicles and on the in vitro metabolism of [(3)H]T to [(3)H]DHT in seminal vesicles homogenates of gonadectomized male hamsters. All trienones 30, 31, and 43-46 in all biological models showed consistently a higher 5alpha reductase inhibitory activity than the corresponding dienones 27, 32, 33 and 38 42. We believe that with these compounds the 5alpha-reductase enzyme is inactivated by an irreversible Michael type addition of the nucleophilic portion of the enzyme to the conjugated double bond of the steroid. The trienones having a more coplanar structure react faster with the enzyme and thus show a higher inhibitory activity. PMID- 12570839 TI - Current strategies for the development of novel antipsychotic drugs. AB - While classical neuroleptics are characterized by dopamine D(2) antagonism, this is also considered to be the cause of their neurological side effects. In recent years, novel antipsychotic drugs with improved efficacy, devoid of extrapyramidal effects are being developed. The mechanisms of action of these new atypical antipsychotics can be classified into three general groups: a) binding to D(2) together with non-dopaminergic receptors, b) interaction with dopamine receptor subtypes other than D(2) and c) selective binding to non-dopaminergic systems, such as glutamatergic, sigma, neurotensin, and cannabinoid. PMID- 12570840 TI - Biological properties and therapeutic potential of bilirubin. AB - Bilirubin was long considered a useless metabolite of heme catabolism, responsible for the clinical manifestation of jaundice, and potentially toxic in high doses, particularly in neonates. In the past two decades the potent biological properties of bilirubin, particularly as an antioxidant, have been recognised, and this has prompted a number of investigations into this molecule concerning its in vitro and in vivo properties. This review summarises that work, as well as more recent investigations into the potential therapeutic uses of bilirubin. PMID- 12570841 TI - 9-hydroxyellipticine and derivatives as chemotherapy agents. AB - The hydroxy group in 9-hydroxyellipticines increases the apparent affinity for DNA, stabilisation of toposiomerase II-DNA cleavable complex, oxidation to reactive quinone-imine intermediates, phosphorylation of p53 suppressor proteins and cytotoxicity relative to the parent ellipticines. Recent studies have focused on the mechanism of inhibition of phosphorylation of the mutant type of p53 protein, structural characterisation of the drug-DNA complex, the synthesis of carbohydrate derivatives and calculations of physical parameters, including dipole moments, as potential screens to allow identification of new active derivatives. Derivatisation at the 2- and 9-positions has lead to significant improvements in the in vivo activity of the 9-hydroxyellipticine derivatives and has provided important insights into the mechanism of action of these compounds. PMID- 12570842 TI - A synthetic overview of new molecules with 5-HT1A binding affinities. AB - The present review discusses the synthetic strategies of new ligands exhibiting mainly 5-HT(1A)binding affinities. Specifically we focused our attention in the synthesis of compounds structurally related to arylpiperazine, 2-aminotetralin, and benzopyran derivatives. PMID- 12570843 TI - New progresses in the enantioselective synthesis and biological properties of carbocyclic nucleosides. AB - The recent advances in the chemistry of carbocyclic nucleosides focused on different synthetic approaches that lead to optically pure products as well as a comprehensive overview of their biological properties are discussed. In the latter aspect, molecular recognition of enzymes of pharmacological importance such as: reverse transcriptase, adenosine deaminase, thymidine kinase, DNA cytosine-C5 methyl transferase, S-adenosylhomocysteine hydrolase, etc are considered. The role of conformation and puckering of the glycon moiety in modulating the biological activity and also the use of carbanucleosides as building blocks to prepare oligonucleotides are carefully illustrated. PMID- 12570844 TI - Inhibitors of lysine biosynthesis as antibacterial agents. AB - Bacterial biosynthesis of lysine has come under increased scrutiny as a target for novel antibacterial agents as it provides both lysine for protein synthesis and meso-diaminopimelate for construction of the bacterial peptidoglycan cell wall. Recent studies of the enzymes of the lysine biosynthetic pathway, development of inhibitors and investigations of their antibacterial properties are discussed. PMID- 12570845 TI - Monoamine oxidases: to inhibit or not to inhibit. AB - Monoamine oxidase (MAO) inhibitors were developed as antidepressants but many drugs, including the novel oxazolidinone antibacterial agents, share similar molecular properties and have MAO inhibitory activity. Factors important for binding antidepressants and modifications to decrease binding of oxazolidinones to avoid undesirable vascular effects are discussed. PMID- 12570846 TI - Opioid ligands having delayed long-term antagonist activity: potential pharmacotherapies for opioid abuse. AB - Buprenorphine is a partial agonist at the micro -opioid receptor with long duration of action and also exhibits delayed antagonist activity. Buprenorphine is finding increasing use as a treatment agent for opioid abuse, though its low efficacy is not well tolerated by all addicts. There is interest in developing a higher efficacy version of buprenorphine and in this mini-review some of the ligands recently discovered, that share with buprenorphine a profile of agonism followed by delayed antagonism, are discussed. PMID- 12570847 TI - Platelet ADP receptors and their antagonists. AB - ADP plays a crucial role in haemostasis and thrombosis and its receptors are potential target for antithrombotic drugs. The knowledge of the ADP-receptors has been amplified by recent discoveries. This review highlights the ADP-receptors models and their antagonists described in the recent literature, mainly the thienopyridine derivatives. PMID- 12570848 TI - Epothilone B and its analogs - a new family of anticancer agents. AB - Epothilones are naturally occurring 16-membered macrolides with the ability to promote tubulin polymerization in vitro and to stabilize preformed microtubules against Ca(2+)- or cold-induced depolymerization. In contrast to paclitaxel (Taxol((R))) epothilones are also active in vitro against multidrug-resistant cancer cell lines as well as cell lines whose paclitaxel-resistance is derived from specific beta-tubulin mutations. Based on their attractive in vitro biological profile epothilones have turned into important lead structures in anticancer drug discovery and hundreds of analogs and derivatives of epothilone A and B have been prepared and biologically characterized over the past four years. A number of compounds, including natural epothilone B, deoxyepothilone B, and epothilone B lactam (BMS-247550) have also been reported to exhibit profound in vivo antitumor activity in animal models. Apart from providing a brief summary of the SAR that has emerged from the above in vitro studies, this minireview will largely focus on the biology and chemistry of those analogs for which in vivo antitumor activity has been reported in the literature. Two of these compounds, natural epothilone B and epothilone B lactam (BMS-247550) have advanced to clinical studies in humans. PMID- 12570849 TI - Naturally occurring peroxides with biological activities. AB - New natural peroxides that have potent biological activities with novel diverse structures are reviewed with classification as secondary metabolites such as terpenes, polyketides, phenolics, and hydroperoxides. These compounds, isolated mainly from medicinal plants and marine sponges, are valuable sources in the drug discovery for particularly antitumor and antimalarial agents. PMID- 12570850 TI - Did quadruplex DNA play a role in the evolution of the eukaryotic linear chromosome? AB - The current evidence on prokaryotic linear chromosomes, the eukaryotes that do not use telomerase and quadruplex DNA has been considered. This has lead to the suggestion that quadruplex DNA may have played a role in the evolution of the protection linear chromosomes rather than in overcoming the end replication problem. PMID- 12570851 TI - The design of G-quadruplex ligands as telomerase inhibitors. AB - Guanine-rich repetitive DNA sequences are of particular importance at the ends of chromosomes, where they are associated with a number of proteins to form telomeres. Their function is in large part to protect chromosomal ends from unwanted degradation and chromosomal fusions, although in normal somatic cells telomeres progressively shorten, eventually becoming non-proliferating and consequently these cells have a finite lifetime. By contrast tumour cell telomeres are maintained in length so that tumour cells are effectively immortalised. The reverse transcriptase enzyme telomerase is activated in over 80% of tumour cells, and it undertakes the synthesis of further telomeric DNA repeats, so directly maintaining telomeres. The inhibition of telomerase leads to the senescence and eventual apoptosis of tumour cells, and thus telomerase is an attractive target for selective chemotherapy. This review describes an approach to the inhibition of telomerase that involves the folding of telomeric DNA into a four-stranded quadruplex structure, held together by Hoogsteen hydrogen-bonded arrays of guanine bases. The formation of a quadruplex structure at the 3' end of telomeric DNA effectively hinders telomerase from adding further repeats. A number of small-molecule ligands are described that stabilise quadruplex formation, and which result in telomerase inhibition. Implications for anti tumour therapy with such molecules are discussed, and the particular challenges and problems discussed. PMID- 12570852 TI - Nucleic acids as targets for antitelomerase agents. AB - Telomeric DNA progressively erodes with each round of cell division in cells that do not express telomerase, a specialized reverse transcriptase necessary to fully duplicate the chromosomal ends. Telomerase is expressed in tumor cells but not in most somatic cells and thus telomeres and telomerase may be proposed as attractive targets for the discovery of new anticancer agents. In this paper we will present different strategies to inhibit telomerase activity via an interaction with a telomere/telomerase nucleic acid component, with a special emphasis on quadruplex ligands. PMID- 12570853 TI - Alternative approaches to the discovery and development of telomerase-targeted anticancer drugs. AB - Four different approaches have been reviewed herein: i) nucleoside analogs as mock agents of the reverse transcriptase (hTERT) catalytic site; ii) miscellaneous molecules with unknown mechanism(s) of action; iii) inhibitors of upstream processes of regulation of the hTERT subunit; iiii) immunotherapy against immunogenic hTERT-derived peptides. PMID- 12570854 TI - Approaches for the inhibition of human telomerase based on the use of peptide nucleic acids and hammerhead ribozymes. AB - The ability of peptide nucleic acids and hammerhead ribozymes, which target different subunits of human telomerase, to efficiently inhibit the enzyme's catalytic activity has been clearly demonstrated in several in vitro studies carried out in human immortalized and cancer cell lines. However, the actual efficacy of these molecules still needs to be validated in in vivo human tumor models, and such validation appears to be largely dependent on the development of reliable systems for their intracellular delivery. PMID- 12570855 TI - Small-molecule inhibitors of actin dynamics and cell motility. AB - Cell motility is a central feature of a range of normal and pathological processes, including embryonic development, tissue repair, immune cell function, angiogenesis, and cancer metastasis. The dynamics of the actin cytoskeleton power cell migration. A large number of proteins are known or suspected to play roles in regulating actin dynamics. While there are now many available small molecules that target the actin cytoskeleton directly, there is a paucity of specific inhibitors of actin-binding proteins and other immediate regulators of actin dynamics and cell movement. This makes the field of exceptional interest as a meeting place between the goals of chemical biology and the needs of cell biology. Furthermore, while regulators of the cell cycle have been recognized for some time as targets for anti-cancer drug development, controlling actin dynamics and cell motility as a therapeutic approach has received scant attention in comparison until recently. This review deals with small-molecule inhibitors of actin dynamics as they relate to cell shape change and motility, from compounds targeting actin directly to those targeting proteins involved in the fundamental control of the actin cytoskeleton. PMID- 12570856 TI - Recent advances in glycomics and glycogenetics. AB - As the human genome sequence is nearly deciphered, it is important to turn the attention to the physiological functions of the genes. Thus, the study of the gene products, the proteins, is the next big challenge. The proteins, however, are not the final gene products in many cases. It has been shown that carbohydrates participate in post-translational modifications and in many other functional regulations, hence the study of the glycome, the entire collection of carbohydrates is essential in order to determine the functions of all genes, and will greatly enhance the field of chemical genetics. Known biological function / targets of carbohydrates and combinatorial synthesis & structural analysis of natural / non-natural carbohydrates are surveyed in this review. Methods to search for new biological targets that include carbohydrate mimetics and carbohydrate scaffolds along with chip technology, are also presented. PMID- 12570857 TI - Custom DNA-binding proteins and artificial transcription factors. AB - Expression of the genome is primarily regulated at the level of transcription by gene-specific transcription factors, which recognize specific DNA sequences to activate or inhibit transcription. The ability to control gene expression at will would provide scientists with a powerful tool for biotechnology and drug discovery research. Over the last decade or so, researchers have made great strides in our understanding of the structures and mechanisms of action of naturally occurring transcription factors. Such research has revealed that members of the Cys2-His2 zinc finger family of transcription factors consist of functional modules that recognize a wide variety of DNA sequences. This review describes recent advances in the development of novel methods to design and construct artificial transcription factors to control gene expression at will. The applications of artificial transcription factors in the areas of medicine and biotechnology are discussed. PMID- 12570858 TI - Supertargeted chemistry: identifying relationships between molecular structures and their sub-cellular distribution. AB - Supertargeted chemistry is the study of how chemical structures localize or direct molecules to specific sub-cellular compartments in living cells. Supertargeting can be used to increase the activity or specificity of an inhibitor against its target, by concentrating the inhibitor in the particular organelle where the target is active. But, unlike structure-activity relationships, structure-localization relationships are not a simple function of compound concentration. Various aspects of mitochondrial physiology, proteomics and pharmacology have made this the organelle of choice for supertargeting studies. While exploration of supertargeting strategies to this and the other organelles has been limited, combinatorial chemical libraries of fluorescent molecules are beginning to illuminate new supertargeting mechanisms at the sub cellular level. Moreover, predictive approaches that determine the relationship between a molecule's features and sub-cellular localization are being developed in the related field of functional genomics. Applied to the small molecules, such strategies could prove useful for predicting structure-localization relationships amongst large libraries of compounds. PMID- 12570859 TI - Micro- and nanotechnologies for studying cellular function. AB - The study of complex biological systems requires methods to perturb the system in complex yet controlled ways to elucidate mechanisms and dynamic interactions, and to recreate in vivo conditions in flexible in vitro set-ups. This paper reviews recent advances in the use of micro- and nanotechnologies in the study of complex biological systems and the advantages they provide in these two areas. Particularly useful for controlling the chemical and mechanical microenvironments of cells is a set of techniques called soft lithography, whereby elastomeric materials are used to transfer and generate micro- and nanoscale patterns. Examples of some of the capabilities of soft lithography include the use of elastomeric stamps to generate micropatterns of protein and the use of elastomeric channels to localize chemicals with subcellular spatial resolutions. These types of biological micro- and nanotechnologies combined with mathematical modeling will propel our understandings of cellular and subcellular physiology to new heights. PMID- 12570860 TI - Array-based technologies and their applications in proteomics. AB - Latest microarray-based technologies, including small molecule-, peptide-, protein- and cell-based arrays, and their applications in the field of proteomics are reviewed. PMID- 12570861 TI - Embryonic stem cells: a perfect marriage between gene regulation and regenerative medicine. AB - The mechanism of mammalian gene regulation is highly complex, involving multiple layers of feedback control loops and dynamic chromatin remodeling. The current approach used to dissect the genetic circuitry of mammalian gene regulation utilizes somatic cells and protein fusion as a means to modulate protein interactions. This approach has several limitations that include (i) genome inaccessibility, (ii) high background interferences and, (iii) limited cellular phenotypes. Previously, the two broad fields of research "control of gene expression" and "stem cell biology" had been pursued separately by cell biologists; this review outlines evidence suggesting that integration of these two fields would provide a comprehensive platform for interdisciplinary research seeking to address mechanistic questions concerning gene regulation-that could have enormous implication for the development of therapeutic applications. PMID- 12570862 TI - Naturally occurring iminosugars and related compounds: structure, distribution, and biological activity. AB - Alkaloids mimicking sugars in size and shape are now believed to be widespread in plants and microorganisms. Iminosugars are monosaccharide analogs in which the ring oxygen has been replaced by an imino group. Such iminosugars inhibit the glycosidases involved in a wide range of important biological processes because of their structural resemblance to the sugar moiety of the natural substrate and the presence of the nitrogen atom mimicking the positive charge of the glycosyl cation intermediate in the enzyme-catalyzed glycoside hydrolysis. These iminosugars and their derivatives are arousing considerable attention as potential therapeutic agents. In this review, the distribution of naturally occurring iminosugars and their biological activities and therapeutic applications will be reviewed and the prospects of iminosugars and their derivatives for new therapeutic applications will also be described. PMID- 12570863 TI - Iminosugars and relatives as antiviral and potential anti-infective agents. AB - Iminosugars have emerged in the literature during mid-1960's as synthetic compounds. Around the same time, the first examples were found in Nature and their interesting enzyme inhibitory properties were recognised. Due to their powerful interference with glycohydrolases (glycosidases) as well as glycosyltransferases, quite a few representatives exhibit notable biological activities. These range from anti-diabetic, insect antifeedant, nematicidal, plant growth regulating to immunomodulating, anti-cancer as well as - in selected cases - anti-infective properties. The latter will be the focus of the following survey. PMID- 12570864 TI - Imino-sugar-based nucleosides. AB - Continued interest in imino-sugar nucleosides derives from the observations that N-ribosyltransferases are powerfully inhibited by imino-C-nucleoside analogues that mimic the ribooxacarbenium ion nature of their transition states. The synthesis of such compounds is challenging, and synthetic routes to imino- and imino-C-nucleosides are reviewed, together with the nature of the N ribosyltransferases that are targets for these analogues. Examples are given to exemplify the promise these compounds hold for the treatments of biological disorders. PMID- 12570865 TI - Design, synthesis and biological evaluation of iminosugar-based glycosyltransferase inhibitors. AB - Due to their pivotal role in the biosynthesis of oligosaccharides and glycoconjugates, glycosyltransferases represent targets of choice for the development of potent inhibitors and therefore for drug discovery. Since glycosyltransferase reactions are thought to proceed through transition states similar to those of glycosidases, iminosugars and the related compounds have been recently used to design potential inhibitors of this major class of enzymes. The purpose of this review is to stimulate further research in this promising area by providing a description and a biological evaluation of the different types of inhibitors containing an iminosugar. These data in addition to the recent insights into the structural basis of glycosyltransferase catalysis may constitute the first steps toward a rational design of potent glycosyltransferases inhibitors. PMID- 12570866 TI - Therapeutic applications of imino sugars in lysosomal storage disorders. AB - The N-alkylated imino sugars have inhibitory activity against the first enzyme in the pathway for glucosylating sphingolipid in eukaryotic cells, ceramide-specific glucosyltransferase. A therapeutic approach termed 'substrate deprivation' or 'substrate reduction therapy' (SRT) aims to reduce biosynthetic capability in the cell to match the reduced lysosomal catalytic activity seen in lysosomal storage disorders. The use of N-alkylated imino sugars to establish this therapeutic strategy is described in cell culture and gene knockout mouse disease models. One imino sugar, N-butyl-DNJ (NB-DNJ) has been in clinical trials for type 1 Gaucher disease and has shown to be an effective therapy for this disorder. PMID- 12570867 TI - gem-Diamine 1-N-iminosugars and related iminosugars, candidate of therapeutic agents for tumor metastasis. AB - Cancer metastasis is one of the major challenges in cancer research. Inhibitors of tumor metastasis are rapidly emerging as important new drug candidates for cancer therapy. Tumor metastasis formation occurs via a complex multistage process which involves a crucial step of tumor invasion through the basement membrane. Tumor cell invasion involves attachment of tumor cell to the basement membrane through laminin, degradation of the matrix by proteolytic enzymes from the tumor cell and cell migration through the basement membrane. New drugs aimed at the metabolism of tumor cell surface oligosaccharides and/or catabolism of glycoconjugates of extracellular matrix and basement membrane could inhibit tumor metastasis. In this article, current progress in the control of tumor metastasis by gem-diamine 1-N-iminosugars and related iminosugars (nojirimycin and d-glucaro delta-lactam), which are potent and specific inhibitors of carbohydrate metabolism and catabolism, has been reviewed. gem-Diamine 1-N-iminosugars related to d-glucuronic acid and l-iduronic acid, nojirimycin and d-glucaro-delta-lactam suppress invasion of B16 melanoma variants and 3LL (lung carcinoma) cells through reconstituted basement membrane, and inhibit pulmonary metastasis of these tumor cells in mice and/or cKDH-8/11 (liver carcinoma) cells in rats. These results suggest that the metabolism of beta-d-glucuronide and alpha-l-iduronide of glycoconjugates and/or the processing of carbohydrates of tumor cell surface may participate in tumor metastasis. That these gem-diamine 1-N-iminosugars and related iminosugars are potent inhibitors of tumor metastasis holds promise of new drug candidates for cancer chemotherapy. PMID- 12570868 TI - Coronary and sympathetic responses to core hypothermia: answers and questions. PMID- 12570869 TI - Gene therapy for ocular angiogenesis. AB - Ocular neovascularization is a central feature of diabetic retinopathy and age related macular degeneration. These conditions are the major causes of blindness in the developed world. Current treatments are of limited efficacy and associated with significant adverse effects. Characterization of the molecular and cellular events involved in angiogenesis has led to the identification of a number of angiostatic molecules with potential therapeutic value. The systemic administration of small molecule angiostatic proteins risks significant systemic adverse effects and the effect of their intraocular injection is short-lived. Local gene transfer, however, offers the possibility of targeted, sustained and regulatable delivery of angiostatic proteins to the retina after a single procedure to introduce a vector to an intraocular site. The effect of intra ocular delivery of recombinant viruses carrying genes encoding angiostatic proteins has been demonstrated in rodent models of ocular neovascularization. Recombinant adeno-associated virus-mediated local gene transfer of a vascular endothelial growth factor inhibitor controls both retinal and choroidal neovascularization. The clinical application of this approach may require the means to regulate gene expression in order to minimize the potential for adverse effects. Regulation of transgene expression by means of a hypoxia-responsive promoter offers an attractive strategy for the targeted and regulated delivery of angiostatic proteins to the retina in the management of ischaemia-induced ocular neovascularization. Preclinical studies of gene transfer in a large animal model following subretinal delivery of a recombinant adeno-associated virus vector have demonstrated efficient sustained reporter gene expression in cells of the outer retina. Recent progress has enabled the planning of clinical trials of gene therapy for ocular neovascular disorders. PMID- 12570870 TI - Circulating adhesion molecules are correlated with ultrasonic assessment of carotid plaques. AB - The relationship between levels of circulating intercellular cell-adhesion molecule-1 (cICAM-1) or P-selectin (cP-selectin) and the severity of carotid atherosclerosis was examined in 301 outpatients undergoing duplex ultrasonographic examination. Carotid plaque was defined as an intima-media thickness greater than 1.0 mm, and a plaque score (PS) was calculated from the plaque thickness in both carotid arteries. Multivariate analysis demonstrated significant positive associations between cICAM-1 and the number of plaques [beta = 0.11; confidence interval (CI), 0.007-0.213], maximum intima-media thickness (beta = 0.11; CI, 0.01-0.219), and PS (beta = 0.10; CI, 0.001-0.205). In contrast, no significant association was found for cP-selectin. cP-selectin did not increase until atherosclerosis was advanced (PS > 10), showing a marked increase in patients with >/= 50% stenosis. The circulating levels of both proteins are related to real measurements of plaque formation in the carotid arteries independently of classical risk factors. Marked elevation of cP-selectin occurs in advanced carotid atherosclerosis after gradual elevation of cICAM-1. PMID- 12570871 TI - MIM-B, a putative metastasis suppressor protein, binds to actin and to protein tyrosine phosphatase delta. AB - We have found that MIM-B, a putative metastasis suppressor protein, is implicated in actin cytoskeletal control and interaction with a protein tyrosine phosphatase (PTP). MIM was originally described as a protein whose mRNA was Missing in Metastasis, as it was found not to be present in metastatic bladder carcinoma cell lines [Lee, Y. G., Macoska, J. A., Korenchuk, S. and Pienta, K. J. (2002) Neoplasia 4, 291-294]. We further characterized a variant of MIM, which we call MIM-B, and which we believe may be a link between tyrosine kinase signalling and the actin cytoskeleton. We have shown, using purified proteins and cell extracts, that MIM-B is an actin-binding protein, probably via a WASP (Wiskott-Aldrich syndrome protein)-homology 2 domain at its C-terminus. We have also found that MIM-B binds to the cytoplasmic domain of receptor PTPdelta. Expression of full length MIM-B induces actin-rich protrusions resembling microspikes and lamellipodia at the plasma membrane and promotes disassembly of actin stress fibres. The C-terminal portion of MIM-B is localized in the cytoplasm and does not affect the actin cytoskeleton when expressed, while the N-terminal portion localizes to internal vesicles and probably targets the protein to membranes. We postulate that MIM-B may be a regulator of actin assembly downstream of tyrosine kinase signalling and that this activity may explain the involvement of MIM in the metastasis of cancer cells. PMID- 12570872 TI - Different binding motifs in metabotropic glutamate receptor type 7b for filamin A, protein phosphatase 1C, protein interacting with protein kinase C (PICK) 1 and syntenin allow the formation of multimeric protein complexes. AB - Metabotropic glutamate receptor (mGluR) type 7-mediated neurotransmission depends critically on its regulation by associated molecules, such as kinases, phosphatases and structural proteins. The splice variants mGluR7a and mGluR7b are defined by different intracellular C-termini, and simultaneous or exclusive binding of interacting proteins to these domains modulates mGluR7-mediated signalling. However, molecular determinants defining binding regions for associated proteins within mGluR7 C-termini are mostly unknown. In the present study, we have mapped the binding domains of four proteins [filamin A, protein phosphatase (PP) 1C, protein interacting with protein kinase C (PICK) 1 and syntenin] interacting with the mGluR7b variant, and show that the alternatively spliced distal part of the mGluR7b C-terminus was sufficient for the interactions. By individual substitution of all mGluR7b isoform-specific amino acids with alanine and construction of a series of deletion constructs, residues important for the interactions were identified and binding regions could be defined. Interestingly, mGluR7b contains an unusual PP1C-binding motif, located at the N-terminus of the binding domains for PICK1 and syntenin. Consistently, binding of PP1C and PICK1 or PP1C and syntenin to mGluR7b was not competitive. Furthermore, PICK1, but not PP1C, interacted physically with syntenin. Our results represent a molecular description of the binding mechanisms of four mGluR7-associated proteins, and indicate the formation of ternary protein complexes composed of mGluR7b, PP1C, PICK1 and syntenin. PMID- 12570873 TI - Calcium and domain interactions contribute to the thermostability of domains of the multimodular cellobiohydrolase, CbhA, a subunit of the Clostridium thermocellum cellulosome. AB - Each of three internal domains of multi-modular cellobiohydrolase CbhA from Clostridium thermocellum, X1(1), X1(2) (previously designated as fibronectin type 3-like modules, Fn3(1) and Fn3(2)) and family 3 carbohydrate-binding module (CBM3) binds 1 mol of Ca(2+). Structures and thermal stabilities of X1(1), X1(2), CBM3, X1(1)X1(2), and X1(1)X1(2)-CBM3 containing Ca(2+) (holo-proteins) and without Ca(2+) (apo-proteins) have been studied using CD spectroscopy. All domains are beta-proteins with irregular far-UV CD spectra due to the aromatic side chain contributions. The positive signal at 294 nm in the near-UV CD spectrum of X1(1) lacking a tryptophan residue might be attributed to the presence of aromatic clusters. Thermal denaturation of all proteins is reversible and results in the total loss of tertiary structure and preservation of significant amount of ordered secondary structure. Removal of Ca(2+) destabilizes polypeptides in a different way and to a different extent. It decreases the melting temperature ( T (m)) (by 20 degrees C) and co-operativity of thermal transition of X1(1), increases the number of transitions and lowers the co operativity of unfolding of CBM3, and slightly decreases T (m)s (2.4-4.2 degrees C) of X1(2), X1(1)X1(2), and X1(1)X1(2)-CBM3. Transitions of X1(1)X1(2) and X1(1)X1(2)-CBM3 follow a two-state model regardless of the presence of Ca(2+). X1(1) is strongly stabilized in the apo-X1(1)X1(2) and apo-X1(1)X1(2)-CBM3 as they display T (m)s similar to those of individual and combined holo-modules. Observed CD spectra of X1(1)X1(2) and X1(1)X1(2)-CBM3 differ from those calculated as the simple weighted sum of individual modules. These differences are more prominent in spectra of apo-proteins. The results indicate the presence of inter-domain interactions in CbhA. Holo-modules, i.e. containing Ca(2+), behave essentially independently, but in the absence of Ca(2+) domain interactions are more important for the conformation of the polypeptides. PMID- 12570874 TI - Curcumin activates the haem oxygenase-1 gene via regulation of Nrf2 and the antioxidant-responsive element. AB - The transcription factor Nrf2, which normally exists in an inactive state as a consequence of binding to a cytoskeleton-associated protein Keap1, can be activated by redox-dependent stimuli. Alteration of the Nrf2-Keap1 interaction enables Nrf2 to translocate to the nucleus, bind to the antioxidant-responsive element (ARE) and initiate the transcription of genes coding for detoxifying enzymes and cytoprotective proteins. This response is also triggered by a class of electrophilic compounds including polyphenols and plant-derived constituents. Recently, the natural antioxidants curcumin and caffeic acid phenethyl ester (CAPE) have been identified as potent inducers of haem oxygenase-1 (HO-1), a redox-sensitive inducible protein that provides protection against various forms of stress. Here, we show that in renal epithelial cells both curcumin and CAPE stimulate the expression of Nrf2 in a concentration- and time-dependent manner. This effect was associated with a significant increase in HO-1 protein expression and haem oxygenase activity. From several lines of investigation we also report that curcumin (and, by inference, CAPE) stimulates ho-1 gene activity by promoting inactivation of the Nrf2-Keap1 complex, leading to increased Nrf2 binding to the resident ho-1 AREs. Moreover, using antibodies and specific inhibitors of the mitogen-activated protein kinase (MAPK) pathways, we provide data implicating p38 MAPK in curcumin-mediated ho-1 induction. Taken together, these results demonstrate that induction of HO-1 by curcumin and CAPE requires the activation of the Nrf2/ARE pathway. PMID- 12570875 TI - Selective interaction of LAT (linker of activated T cells) with the open-active form of Lck in lipid rafts reveals a new mechanism for the regulation of Lck in T cells. AB - In T cells, the lipid raft-associated Lck is strongly tyrosine phosphorylated and has reduced enzymic activity in contrast with the detergent-soluble pool, which has substantial activity. Lck tagged at the C-terminus (Lck/V5-His) was efficiently captured by epitope-specific reagents from the detergent-soluble fraction but not from lipid rafts. Binding was restored following urea denaturation, suggesting that Lck/V5-His is in a 'closed' conformation in these domains. In agreement with this hypothesis, the Tyr(505) --> Phe/V5-His and Arg(154) --> Lys/V5-His mutants, which disrupt the SH2-Tyr(505) intramolecular interaction, were efficiently precipitated from lipid rafts. In contrast to Lck, Fyn/V5-His was precipitated equally well from both fractions. In the LAT(linker of activated T cells)-deficient J.CaM2 cells, Tyr(505) phosphorylation of raft associated Lck was reduced whereas its enzymic activity was elevated. This correlated with decreased levels of raft-localized Csk (C-terminal Src kinase) kinase. Increased tyrosine phosphorylation of Lck was restored in LAT reconstituted J.CaM2 cells suggesting that LAT negatively regulates Lck activity in lipid rafts. Co-immunoprecipitation experiments from Tyr(505) --> Phe/V5-His expressing cells revealed that LAT preferentially interacts with the 'open' form of Lck in T cell raft domains. These results demonstrate that, unlike the non raft pool, Lck in lipid rafts has a 'closed'-inactive structure, and that LAT plays a role in maintaining this conformation, possibly by facilitating critical associations within lipid rafts via its capacity to interact with the 'open' form of the kinase. PMID- 12570876 TI - No bias of ignored bilaterality when analysing the revision risk of knee prostheses: analysis of a population based sample of 44,590 patients with 55,298 knee prostheses from the national Swedish Knee Arthroplasty Register. AB - BACKGROUND: The current practice of the Swedish Knee Register is not to take into consideration if one or both knees in a patient are subject to surgery when evaluating risk of revision after arthroplasty. Risk calculations are typically done by statistical methods, such as Kaplan-Meier analyses and Cox's proportional hazards models, that are based on the assumption that observed events are independent, and this is rarely appreciated. The purpose of this study was to investigate if ignoring bilateral operations when using these methods biases the results. METHODS: The bias of not taking bilateral operations into account was investigated by statistically analysing 55 298 prostheses in 44 590 patients, undergoing knee arthroplasty surgery in Sweden during 1985-1999, using traditional proportional hazards analysis, which assumes that all observations are independent, and a shared gamma frailty model, which allows patients to contribute repeated observations. RESULTS: The effect of neglecting bilateral prostheses is minute, possibly because bilateral prosthesis failure is a rare event. CONCLUSION: We conclude that the revision risk of knee prostheses in general can be analysed without consideration for subject dependency, at least in study populations with a relatively low proportion of subjects having experienced bilateral revisions. PMID- 12570877 TI - The 2002 World Summit on Sustainable Development: a more holistic approach for public health nutrition? PMID- 12570878 TI - The Indaba declaration on food, nutrition, health and sustainable development. PMID- 12570879 TI - Breast-feeding and the working mother in Nairobi. AB - OBJECTIVE: In the fast growing city of Nairobi, women often combine the roles of mother and worker in trying to achieve better standards of living. The objective of this study was to document the effect of returning to work on breast-feeding by mothers in Kenya. DESIGN: A cross-sectional survey. SETTING: Outpatient clinics of two major hospitals in Nairobi, one government hospital in an economically deprived area and one high-fee private hospital. SUBJECTS: Four hundred and forty-four working mothers from low and higher socio-economic areas in Nairobi. All working mothers with infants aged 4 to 12 months attending during the survey period were invited to participate. RESULTS: The prevalence of breast feeding at the time of interview was found to be 94.1%. The lower socio-economic group exhibited a higher prevalence of breast-feeding (99%), 10% greater than the higher socio-economic group. The mean number of hours the mothers were away from home due to work was 46.2 hours each week. The majority (54.4%) of the mothers employed a 'house-girl' to care for their infant while they were at work, while 28.4% were able to take their infants to work. Most of the breast-feeding mothers (95%) breast-fed their infants at least three times a day and only 23 mothers reported not being able to breast-feed their infants during the day. The lower socio-economic group had a mean of 5.09 breast-feeding times per day while the higher socio-economic group had a mean of 3 times a day. In a logistic regression analysis the mode of work (fixed working hours vs. shift working hours) was associated with exclusive breast-feeding at one month (odds ratio (OR)=0.45) and two months (OR=0.39). CONCLUSION: In Western countries 'return to work' is often cited as the reason that breast-feeding is discontinued prematurely. In this study we have shown how mothers in Kenya are able to successfully continue breast feeding after they have returned to work, often for very long hours. PMID- 12570880 TI - Urbanisation and cariogenic food habits among 4-24-month-old black South African children in rural and urban areas. AB - OBJECTIVE: To determine if social class, education level and group environment (rural and urban) influence particular food habits commonly associated with dental caries incidence among 4-24-month-old black South African children. DESIGN, SETTING AND SUBJECTS: Information was collected by trained interviewers using a food-frequency questionnaire from mothers of children in two areas in South Africa: Ndunakazi, a rural area in KwaZulu/Natal (n = 105) and two urban areas in Gauteng - Soweto (low to middle socio-economic area) (n = 100) and the northern suburbs of Johannesburg and Sandton (middle to upper socio-economic area) (n = 101). Education level and occupation of the parents, which define social class, were also recorded. A linear logistic (Proc Catmod) analysis tested social class, education level and group environment as the independent variables and the food habits as the dependent variables. RESULTS: Group environment was significantly associated with nine of the 18 food habits investigated. More urban than rural mothers added sugar to their child's comforter. More mothers in urban Soweto than in urban Johannesburg were still breast-feeding their infants at 24 months. More rural than urban mothers were giving 'mutis' (common and traditional medicines). Together with group environment, education level was significantly associated with giving of 'mutis' and the frequency of giving them. Social class was significantly associated with the frequency of breast-feeding and when the child was breast-fed. Mothers from the upper social class breast-fed less frequently than mothers from the lower class. CONCLUSION: The study showed a strong influence of rural/urban environment on specific cariogenic food habits among young black South African children, enabling the development and implementation of a nutrition strategy. PMID- 12570881 TI - Diarrhoea and child feeding practices in Saudi Arabia. AB - OBJECTIVE: The aim of the study was to estimate the prevalence of diarrhoea in children less than two years old and study the relationship between diarrhoeal episodes and action taken for these episodes by their mothers. DESIGN: The prevalence of diarrhoeal episodes among children and its associations with sociodemographic information and anthropometric measurements of the subjects was examined. Predictive factors for morbidity-associated diarrhoeal disease and actions taken for this were explored. SETTING: Primary health care centres (PHCCs) in Riyadh, Kingdom of Saudi Arabia. SUBJECTS: Children less than two years of age. RESULTS: Nearly a quarter of the children contracted diarrhoea during the two weeks preceding the data collection point, giving about six episodes of diarrhoea per child per year. Diarrhoea was more common in children over 6 months of age, in children who had no vaccination or follow-up cards, and in those who were taken care of by friends and neighbours if their mothers were working outside the home. The mothers of the affected children were young, married before 25 years of age with 2-6 years of formal schooling. During diarrhoeal episodes, about 25% of mothers stopped or decreased breast-feeding, 11.3% reduced the volume of fluids given to their children, and 22.7% of children were fed less solid/semi-solid foods. Mothers used oral rehydration salt in more than 40% of diarrhoeal episodes and unprescribed antibiotics were used in 17% of cases. The mothers who were not taking appropriate action included young mothers with low education level and those working outside the home. CONCLUSION: Diarrhoea is common in children less than two years old in Riyadh City, and intervention based in PHCCs needs to be undertaken to correct the faulty practices of mothers during diarrhoeal episodes in their children. Health education messages should emphasise feeding during diarrhoeal episodes. PMID- 12570882 TI - Sociodemographic inequalities in the diet of young children in the 1946 British birth cohort. AB - OBJECTIVE: To evaluate whether there was food and nutrient equality across occupational social classes and geographical region for members of the 1946 British birth cohort at age 4 years. DESIGN: Cross-sectional analysis of selected food groups, energy and nutrients from one-day recall diet records. SETTING: England, Scotland and Wales in 1950. SUBJECTS: Nationally representative sample of 4419 children aged 4 years in 1950 from the MRC National Survey of Health and Development (NSHD) (1946 Birth Cohort). RESULTS: Significant food and nutrient inequalities occurred by region and occupational social class of the father. Disparity in fruit and vegetable consumption primarily led to the nutrient differences, especially with respect to lower vitamin C and carotene intakes in children from Scotland and from a manual social class background. Lower energy intake in Scottish children was attributable to inequality in the consumption of foods providing fat, and also to the retention of the traditional Scottish diet that included porridge and soups. Consumption of some rationed foods -- bacon, orange juice and tea -- was inequitably distributed by father's social class, but others, in particular meat and spreading fats, were consumed more uniformly. In contrast to fruits and vegetables, which showed marked sociodemographic disparities, other non-rationed foods such as bread and potatoes were consumed universally. CONCLUSION: Local cultural norms may have played as strong a part in sociodemographic differences in the diet of children in the early 1950s as did the strict, post-war food rationing that prevailed. In consequence, nutritional equality was not achieved, and the relatively low intake of antioxidant vitamins during early childhood in certain population groups may have compromised health in the long term. PMID- 12570883 TI - The contribution of foods to the dietary lipid profile of a Spanish population. AB - OBJECTIVE: To identify the food that has the greatest effect on the variation in the percentage of energy intake derived from fat and saturated fatty acids for the consumption of a Spanish population. DESIGN: A cross-sectional study of food consumption, using the 24-hour recall method for three non-consecutive days, one of which was a non-working day. Subjects were interviewed by trained interviewers in the subjects' homes. We used multiple linear regression for statistical analysis. SETTING: The citizens of Reus. SUBJECTS: One thousand and sixty subjects over five years old, randomly selected from the population census of Reus. RESULTS: In both sexes, the foods that mainly determine a high consumption of fat are oil and red meat while those that determine a lower consumption of fat are bread, savoury cereals and fruit. The foods that mainly determine a high consumption of saturated fatty acids are red meat and whole-fat dairy products while those that determine a low consumption are bread, savoury cereals and fruit. CONCLUSIONS: In our population, feasible variations in the intake of some foods - less than one portion - would reduce the estimated percentage of energy intake derived from fat and saturated fatty acids by a quantity considered important for cardiovascular disease prevention. The periodic identification and quantification of the food that most affects the dietary fat profile will help in drawing up dietary guidelines with more reasonable strategies for consuming a healthier diet and decreasing the risk of developing nutritional disorders. PMID- 12570884 TI - Diet patterns and the risk of renal cell carcinoma. AB - OBJECTIVES: Our objective was to identify food intake patterns that might be associated with the risk of renal cell carcinoma. DESIGN: A total of 461 cases (210 females, 251 males) were age frequency matched to population controls. Diet factors were created using factor analysis of 69 food items from a food-frequency questionnaire. These factors were modelled using logistic regression to identify those associated with renal cell carcinoma. SETTING: We investigated the role of diet in the aetiology of renal cell carcinoma using a population-based case- control study conducted in Ontario between 1995 and 1996. SUBJECTS: Cases were Ontario residents 20 to 74 years of age identified through review of pathology reports in the Ontario Cancer Registry. RESULTS: A 'dessert' diet factor was positively associated with disease for both sexes (odds ratio estimate (OR) for males = 3.7, 95% confidence interval (CI) 2.0-6.9; OR for females = 1.4, 95% CI 0.8-2.2, for the highest vs. lowest quartile). In males, a 'beef' diet factor was identified and was associated with an increased risk of renal cell carcinoma. Furthermore, a 'juices' diet factor also showed an association with increased risk in males (OR = 1.8, 95% CI 1.0-3.1). For females, a positive association was observed between renal cell carcinoma and an 'unhealthy' diet factor (OR = 1.4, 95% CI 0.8-2.4). CONCLUSIONS: Our findings confirmed that high-fat and high protein diets might be risk factors for renal cell carcinoma. The data also suggest an increased risk associated with juice intake, a finding not previously reported. PMID- 12570885 TI - Validation of the second version of a quantitative food-frequency questionnaire for use in Western Mali. AB - OBJECTIVE: To assess the relative validity of the second version of a quantitative food-frequency questionnaire (QFFQ), designed to measure the habitual food and nutrient intake in one season in rural populations in Western Mali, West Africa. DESIGN: The dietary intake during the previous week was assessed with the 164-item QFFQ administered by interview. This was compared with the intake from a 2-day weighed record (WR) with weighed recipes. SETTING: The village of Ouassala in the Kayes region, Western Mali. SUBJECTS: Thirty-four women and 36 men aged 15-45 years, from 29 households. RESULTS: The QFFQ gave a lower intake of lunch and dinner and a higher intake of snacks than the WR. The discrepancies were larger for women than for men. The median proportion of subjects classified in the same quartile of intake was 29% for food groups and 36% for energy and nutrients. For classification into extreme opposite quartiles, the median proportion was 6% for food groups and 7% for energy and nutrients. Spearman's rank correlation for energy and nutrients ranged from 0.16 (% energy from protein) to 0.62 (retinol equivalents). CONCLUSIONS: The second version of the QFFQ tends to underestimate total food weight. The methods used for estimating food portion size should therefore be applied with caution. The changes made from the first version had little effect. The ability to rank subjects according to dietary intake is similar with both versions. The improved layout of the new QFFQ makes it a more user-friendly tool for comparing dietary intake between population groups and for measuring changes over time. PMID- 12570886 TI - Can the EPIC food-frequency questionnaire be used in adolescent populations? AB - OBJECTIVE: To assess the validity of the food-frequency questionnaire used in the European Prospective Investigation of Cancer (EPIC FFQ) for estimating nutrient intake in an adolescent population. DESIGN: Sixty-seven schoolchildren (mean age: 12.3 +/- 0.3 years) were recruited to complete a 7-day weighed dietary record (7 day WDR), the EPIC FFQ and supply one 24-hour urine collection. SETTING: Harris Academy in Dundee (UK). RESULTS: Fifty subjects completed both dietary assessment methods. Thirteen of these were classified as underreporters with energy intake/basal metabolic rate < 1.14. The EPIC FFQ showed higher estimates than the 7-day WDR for all nutrients. The median Spearman correlation coefficient for the nutrients examined was found to be 0.31 and increased to 0.48 after adjustment for total energy. The limits of agreement were as far apart as 13.4 MJ, 120 g, 270 g, 120 g and 1170 mg for energy, fat, sugar, protein and calcium, respectively. Correlations between urine and 7-day WDR dietary nitrogen and potassium were found to be statistically significant with r = 0.45 (P < 0.05) and r = 0.78 (P < 0.001) respectively. The median proportion of subjects that appeared in the same and opposite third of intake was found to be 45.9% and 10.8%, respectively. CONCLUSIONS: The EPIC FFQ seems adequate to correctly classify low, medium and high consumers and might therefore be used to identify adolescent population groups at risk or for differences between populations. However, agreement between the EPIC FFQ and the 7-day WDR was very poor on both a group and an individual basis, and demonstrates that the EPIC FFQ is not an appropriate method for estimating absolute intakes in this age group. PMID- 12570887 TI - Urinary excretion of dithiocarbamates and self-reported Cruciferous vegetable intake: application of the 'method of triads' to a food-specific biomarker. AB - OBJECTIVE: Greater intake of Cruciferous vegetables (e.g. broccoli) may prevent cancer at several sites. Urinary excretion of isothiocyanate conjugates (dithiocarbamates, DTC) provides a specific biomarker of Cruciferous vegetable consumption suitable for epidemiological investigations. However, no gold standard referent is available for evaluating urinary DTC levels as an estimator of Cruciferous vegetable consumption. We compared urinary DTC levels to intake as measured by two self-reported dietary assessment techniques. DESIGN: Cruciferous vegetable consumption was measured before and after a behavioural dietary intervention using multiple 24-hour recalls (24HR), a food-counting questionnaire (VFQ) and urinary DTC excretion levels. Analysis included a structural equation approach (Method of Triads) combining these three assessment techniques to estimate the relationship between DTC level and the study population's 'true' Cruciferous vegetable intake. SETTING: The intervention curriculum assisted participants in consuming about 2 servings per day for a 6-week period. Participants attended four classes emphasising problem-solving skills, dietary counselling and vegetable preparation skills. There were no dietary restrictions. SUBJECTS: Thirty-three healthy, free-living, post-menopausal women. RESULTS: Although few participants reported Cruciferae consumption prior to the intervention, 30 participants reported Cruciferae consumption after the intervention (Post-intervention). Urinary DTC levels were correlated with estimated intake derived from either the 24HR (r = 0.57; 95% confidence interval (95% CI) 0.28, 0.76) or VFQ (r =0.49; 95% CI 0.17, 0.71). The validity coefficient (Method of Triads) between urinary DTC excretion and an index of true Cruciferous intake was stronger than the Pearson correlation (r(v) = 0.65; 95% CI 0.35, 0.90), and comparable to estimates derived from the 24HR (r(v) = 0.82; 95% CI 0.65, 1.00) or VFQ (r(v) = 0.76; 95% CI 0.47, 0.92) method. These associations were not affected by adjustment for body mass index, energy intake, or social approval or desirability response sets. CONCLUSIONS: Food-frequency questionnaires (FFQ) suitable for large epidemiological studies may not be designed to measure all Cruciferae, and cannot capture exposure to phytochemicals derived from those vegetables. Urinary DTC measurement was significantly correlated with Cruciferae intake derived from two dietary assessment approaches, and urinary DTC levels could supplement traditional FFQ data by providing an index of recent Cruciferous vegetable intake not susceptible to reporting biases. PMID- 12570888 TI - A Diet Quality Index for Pregnancy detects variation in diet and differences by sociodemographic factors. AB - OBJECTIVE: Methods currently used to assess nutritional status during pregnancy have limitations if one wishes to examine the overall quality of the diet. A Diet Quality Index for Pregnancy (DQI-P) was developed to reflect current nutritional recommendations for pregnancy and national dietary guidelines. DESIGN: Dietary intake was assessed during the second trimester using a food-frequency questionnaire. The DQI-P includes eight components: % recommended servings of grains, vegetables and fruits, % recommendations for folate, iron and calcium, % energy from fat, and meal/snack patterning score. Scores can range from 0 to 80; each component contributed 10 points. SETTING: Two public prenatal clinics in central North Carolina. SUBJECTS: N = 2063 pregnant women who participated in the Pregnancy, Infection, and Nutrition (PIN) Study. RESULTS: The DQI-P quantitatively differentiated diets. The mean score for the population was 56.0 (standard deviation 12.0). Women who were >30 years old, >350% of poverty, nulliparous and high school graduates had significantly higher overall DQI-P scores. Higher percentages of recommended vegetable servings were consumed by higher-income, older and better-educated women. Greater percentages of recommended intakes of folate and iron were seen among black, low-income and nulliparous women. Higher iron intakes were also seen among women who graduated high school and were less than 30 years old. Other differences were observed for intake of fat and meal/snack pattern. Because this index was based on national recommendations, the DQI-P may be a useful tool for research and public health settings to evaluating overall diet quality of pregnant women. PMID- 12570889 TI - [Characterization of de novo malignancies in liver transplantation]. AB - INTRODUCTION: The incidence of de novo malignancies after liver transplantation varies from 3-15%, and is greater than that in the general population. Immunosuppression may play a significant role in the development of most of these tumors. OBJECTIVE: To evaluate the incidence and clinical features of de novo tumors in liver transplant recipients in our center as well as to assess survival. PATIENTS AND METHODS: We retrospectively analyzed 437 liver transplantations (380 patients) performed from April 1990 to July 2001. The incidence of de novo malignancies was 7.4% (n = 28). Four patients presented two different tumors during their lifetime. The etiology of the underlying disease was alcoholic cirrhosis (45.8%), hepatitis C virus cirrhosis (20.8%), hepatitis B virus cirrhosis (12.5%), autoimmune disease (8.4%) and other causes (12.5%). The most frequent neoplasms were cutaneous and epidermoid tumors (21.4% of the malignancies both groups). All the patients with epidermoid tumors and adenocarcinomas were active smokers. The mean age at diagnosis was 58 9 years and this was a factor that influenced tumoral type (adenocarcinomas in older patients and epidermoid tumors in younger patients; p = 0.04). RESULTS: Sarcomas and adenocarcinomas appeared sooner after transplantation than epidermoid and cutaneous tumors (p = 0.04). Fifty percent of the malignancies developed in the second and third year after transplantation. The type of immunosuppression did not influence tumoral type, although most patients received cyclosporin A in combination with azathioprine and/or corticoids. The mean duration of follow-up after diagnosis of the tumor was 23.1 28 months (range, 1-81). Mortality was 58.4% with a median survival of 9 16 months. The actuarial probability of survival at 1, 3 and 5 years was 46.1, 27.7 and 27.7%, respectively. CONCLUSIONS: De novo malignancies are frequent after liver transplantation and their clinical course differs from that in the general population. Because their clinical course is more aggressive, regular follow-up of these patients is essential for early diagnosis. PMID- 12570890 TI - [Ulcerative colitis with segmental involvement]. AB - INTRODUCTION: Ulcerative colitis is a chronic inflammatory disease affecting areas of the colon or the full length. From the endoscopic point of view, ulcerative colitis presents lesions that stretch continuously from the rectum to variable colon segments, a characteristic that is of great value when distinguishing it from Crohn's disease. Continuous involvement, without healthy patches, justifies ending endoscopic exploration once the distal end of the lesion has been reached. OBJECTIVE: To retrospectively study the frequency of segmental lesions in the colonoscopies performed in patients with ulcerative colitis. METHODS: Diagnosis of ulcerative colitis and proctitis was established by clinical, endoscopic, histologic, analytical, and radiological criteria. The indication and number of endoscopies was made on the basis of the clinical criteria of diagnosis, acute episodes, refractoriness or dysplasia screening. The extent of the examination also depended on clinical criteria: the severity of the episode, tolerance to colonoscopy or the degree of cleansing. RESULTS: A total of 155 coloscopies were performed. In 113 colonoscopies (73%) the distal end of the lesion was reached and in 70 (45%) the cecum was reached. Of the 80 patients, 27 (33%) presented ulcerative proctitis at diagnosis. Nine of the 80 patients (11.3%) biopsies were performed in healthy colonic patches, which confirmed histological normality. Six of the 9 patients were receiving no treatment. In all patients except two, the cecum was reached in one or more of the colonoscopies. The distribution of the segmental lesions varied but these were mainly found in the periappendicular region and in the cecum in 6 of the 7 patients in whom the cecum was reached. Of the 80 patients, endoscopic evidence of rectal sparing was found in 5 (6.3%); of these, 4 were receiving systemic or topical treatment. Histological analysis confirmed the absence of inflammatory lesions in these patients. The only patient who was not receiving treatment presented microscopic lesions compatible with ulcerative colitis. CONCLUSIONS: Endoscopic segmental lesions in ulcerative colitis were present in 11.3% of patients. Segmental lesions were most frequently found in the cecum and periappendicular region. Endoscopic and histologic evidence of rectal sparing may be the result of systemic or topical treatment. PMID- 12570891 TI - [Guidelines for the diagnosis and treatment of acute non-variceal upper gastrointestinal bleeding]. PMID- 12570892 TI - [Molecular aspects of duodenal iron absorption]. PMID- 12570893 TI - [Research using medication. The regulatory norms must be respected]. PMID- 12570894 TI - [Hospitalizations preventable by timely and effective primary health care]. AB - OBJECTIVE: To know the specific health problems referred as ambulatory care sensitive conditions (ACSC) and to identify the primordial interventions of primary health care (PHC) in reducing hospitalisations due to ACSC. DESIGN: Descriptive study of hospital discharges generated in Catalonia during 1998-1999, and a Delphi study to reach information about PHC primordial interventions. MEASURES: Diagnostic codes of ACSC selected as markers of PHC effectiveness were used. We analysed hospital discharge distribution by age groups and overall, and hospitalisation rates with its 95% confidence intervals. Descriptive analysis of consensus reached by experts using self-administrated questionnaires was done. RESULTS: The 8.42% of total discharges were due to ACSC. The majority of these (86.9%) fell in 4 of the 13 diagnostic categories included in the ACSC list. A great variety of pathologies with different frequencies were identified. Primary prevention and early diagnoses and treatment were considered as primordial interventions. Chronic health problems needed multimodal interventions. CONCLUSIONS: Diagnostic codes included in each diagnostic category were congruent with the diseases identified. Interventions that could prevent hospitalisations due to ACSC are contemplated as role of PHC. Indicator validity to assess PHC effectiveness is maintained by both results. PMID- 12570896 TI - [Do men and women have different perceptions of pain?]. AB - OBJECTIVE: To evaluate possible differences between men and women's perception of a painful stimulus (venous puncture). DESIGN: Transversal, analytic and observational study.Setting. Primary care.Participants. 709 people over 14 who attended three health centres for blood extraction. 98 cases were rejected (most because they received more than one puncture). MAIN MEASUREMENTS: Age, sex, tobacco habit, perception of pain (through visual analogic scale from 1 to 10), complaints expressed (oral and/or by gestures) and patient's prior expectations. RESULTS: Mean age: 47.4; 60.1% women. The intensity of pain had an average of 1.35 (SD, 1.60; mode, 0; median, 0.8). 14.7% expressed a complaint. Median of pain was 0.95 (Q3-Q1, 1.9-0.4) in men, whereas it was 0.70 (Q3-Q1, 1.725-0.3) in women (U=40.48; P=.095). No correlation was found between valuation of pain and age (R=0.055). No differences were found for smokers (median, 0.925; Q3-Q1, 1.725 0.35) or non-smokers (0.75; 1.825-0.325) (P.05). There were statistically significant valuations that varied according to the prior expectations of pain. Equally, pain was assessed as more intense by those who expressed a complaint (median, 2.1; Q3-Q1, 3.85-0.475) versus those who did not (0.625; 1.55-0.3) (P<.001). CONCLUSIONS: We found no significant differences between men and women's perception of painful stimulus. In any case, differences were clinically irrelevant. There is wide individual variability, which suggests that the essential factors are psycho-social. We have to assess and treat properly a patient in pain, regardless of his/her gender. PMID- 12570897 TI - [Minor surgery in primary care: is continuing education within the team a valid strategy for improving quality?]. AB - OBJECTIVE: To evaluate the impact of continuing education within the team (FCI, in Spanish) on the quality of minor surgery. DESIGN: Study of level of quality. SETTING. Primary care. PARTICIPANTS: First evaluation: all the lesions referred for biopsy during 1998 (62 samples). Second evaluation: those referred in 1999 2000 (150). MAIN MEASUREMENTS: Four explicit criteria regulating procedure and result were designed: C1, sufficient information; C2, correct referral; C3, correct extirpation of lesion; C4, clinical-pathological concordance. Request forms and anatomical-pathological reports were assessed. Evaluation was before and after corrective measures (FCI and organisational changes designed to support FCI). The Kappa index of inter-observer concordance, the Compliance Index and Fisher's Z index were analysed. RESULTS: 62 lesions were included in the first evaluation, with high reliability for C1 and C4, good for C2 and moderate for C3. 150 lesions were included in the second evaluation. The compliance indices showed statistically significant increases from the first to the second evaluation for C1 (38.09% and 50.66%, relative improvement of 19%) and C4 (68.85% and 85.2%, relative improvement of 53%). C2 showed an improvement, but without statistical significance (87.30% and 92.66%). The compliance index for C3 dropped (94.73% and 87.50%). 5.33% of cases in the second evaluation (8 biopsies) were malignant or pre-malignant lesions, compared with 20.96% in the first (P<.05). 100% of these latter showed free resection limits. 77.99% of lesions studied through a biopsy in the second evaluation were nevus, seborrhoeic keratosis or dermatofibroma. CONCLUSIONS: FCI is a valid strategy for improving the quality of programmes of minor surgery in primary care. There was significant improvement in the identification of malignant and pre-malignant pathology, in correct referral and in the clinical-pathological concordance of the lesions. PMID- 12570898 TI - [Health services utilization in Mexico: the perspective of individuals with diabetes]. AB - AIMS: This paper reports findings of a qualitative study whose focus is health services use among individuals with chronic illness. DESIGN: Qualitative research with an ethnographic orientation. SETTING: A low income neighborhood of Guadalajara, Mexico. SUBJECTS: Twenty subjects selected through a theoretical sample. Measurements and main results. Open-ended interviews. The data were content-analyzed. Most diabetic sufferers live with two or more chronic illnesses. Medical services use is combined with that of other healing systems. Participants also combine different types of health care services. Four health care use categories were found: frequent, occasional, periodical, and abandonment. Those who use health services frequently have oscillating or deteriorating illness trajectories, are supported by family members and are primarily women and elderly/retired men. Those who abandon health services have stable trajectories, they are their families primary providers, and cannot stop work to receive medical care; but some others have deteriorating illness trajectories and their symptoms do not ameliorate with consultations. CONCLUSIONS: Different factors explain patterns of health services utilization. Such patterns of use have serious implications in the health care reorganization. PMID- 12570899 TI - [Prevalence of psychiatric pathology at a rural health centre]. AB - OBJECTIVES: To find the prevalence of psychiatric pathology at a rural health centre and to compare the results with results from studies with a similar design. DESIGN: Two-stage crossover study of prevalence: first, detection and then, psychiatric pathology evaluation.Setting. El Marquesado Health Centre (Granada). Scattered rural population with low social and economic level and high percentage of elderly people. PARTICIPANTS: Simple randomised sample representing the population over 18. N=245; error alpha=5%. Sampling source: electoral register. MEASUREMENTS: GHQ-28 Goldberg Questionnaire, social and demographic data, and CAGE test. The CIS questionnaire for psychiatry was administered to those who exceeded the cut-off point (GHQ-28=5) and they were allocated a DSM IV diagnosis. If CAGE was>=2, damaging alcohol consumption was studied. If there was a prior diagnosis, the second stage was avoided. Measurements of prevalence with their exact confidence intervals were made. RESULTS: Prevalence of psychiatric pathology: 28.6% (34.5% in women and 22.6% in men). 30% of cases had two diagnoses. Anxiety disorders were the most common pathology (8.97%), representing 23.9% of the grouped diagnoses. Depression disorders had 8.16% prevalence; problematic consumption of substances, 6.12%. The most common isolated diagnosis was Dysthymia (9.78% of cases). Hidden psychiatric morbidity reached 40%. CONCLUSIONS: The prevalence of psychiatric pathology is very high. The most common disorders are those of anxiety, followed by depression and damaging alcohol consumption. PMID- 12570900 TI - [Surgery and minor traumatology. Can we? Will we be able to?]. PMID- 12570901 TI - [Reflections on living wills (I and II)]. PMID- 12570904 TI - [Pneumococcal vaccines: problems and solutions]. PMID- 12570905 TI - [Primary care evaluation and hospitalization due to ambulatory care sensitive conditions. Conceptual framework]. PMID- 12570906 TI - [LADA-type diabetes. A category to be taken into account in primary health care]. PMID- 12570907 TI - [Efficacy of lipid-lowering treatment in the adult population at a health centre]. PMID- 12570908 TI - [My knee has been hurting for a month. Can't you do something for me?]. PMID- 12570909 TI - [Measuring blood pressure: approximate or scientific]. PMID- 12570910 TI - [Mortality register in a health district: experience three years after its introduction]. PMID- 12570912 TI - [Growth hormone deficiency in adults: effects of replacement therapy on body composition and health-related quality of life]. AB - BACKGROUND AND OBJECTIVE: Deficiency of growth hormone (GH) in the adult is accompanied by changes in the body composition and a diminished health-related quality of life (HR-QoL). The aim of the study was to assess the biochemical response to GH replacement therapy and its safety as well as the resulting body composition and HR-QoL. PATIENTS AND METHODS: One hundred sixty-five patients with hypopituitarism and GH deficiency were studied. A double-blind,randomized, placebo-controlled, 6-months study was first designed,then followed by a further 6-months period in which all patients received GH. The initial GH dose was 0.125 IU/Kg/week followed by 0.250 IU/Kg/week. The body composition was determined by bioelectric impedanciometry and the HR-QoL was evaluated by the Nottingham Health Profile (NHP) and the QoL-AGHDA questionnaire. RESULTS: A significant increase in fat-free mass was observed during treatment with GH, which was accompanied with a simultaneous decrease in fat mass. Total body water increased during GH treatment. Energy and emotional reaction areas evaluated by the NHP showed changes at 6 months; no changes were observed in the remaining dimensions. A progressive improvement was observed in the QoL-AGHDA score in the treated group but not in the placebo group. Adverse events mainly consisted of fluid retention which resolved upon decrease of the dose. CONCLUSIONS: GH treatment in GH deficient adults is in general well tolerated and leads to beneficial effects on body composition and HR-QoL. PMID- 12570913 TI - [Stroke code impact on the efficacy of thrombolytic treatment]. AB - BACKGROUND AND OBJECTIVES: Most stroke patients arrive to emergency rooms too late, many times beyond the therapeutic window for thrombolysis. The activation of a Stroke Code (SC)has proven to be effective in decreasing the latency time for starting therapy in clinical trials. However, its usefulness in routine clinical practice is uncertain. The aim of this study was to determine the feasibility of applying an SC in the daily clinical practice and to evaluate its impact on the efficacy of t-PA thrombolytic treatment. PATIENTS AND METHODS: We included middle-cerebral artery stroke patients treated with IV t-PA from June 2000 to December 2001 who were categorized into three periods of 6 months each. The influence of SC activation on the latency time, arterial recanalization and early neurological recovery was evaluated. RESULTS: Forty-eight patients were prospectively studied. A significant increase in the number of t-PA treated patients was observed in the three periods (4.8%, 5.3% and 8%). A significant progressive decrease (p = 0.036) in the overall delay to start t-PA was observed (from 161.4 to 130 minutes). The proportion of t-PA treated patients in whom SC was activated increased gradually (p = 0.032). SC activation reduced time from symptom onset to hospital arrival (p = 0.013) as well as the "door-to-needle"time (p = 0.049). The rate of early recanalization (p = 0.027) and neurological recovery (p = 0.036) at 48 hours was significantly higher in patients in whom SC was activated than in those in whom SC was not. CONCLUSIONS: SC activation is very useful in the management of acute stroke, increasing the rate of t-PA treated patients and improving the clinical outcome. PMID- 12570914 TI - [Validation of the Spanish version of the Brief Pain Inventory in patients with oncological pain]. AB - BACKGROUND AND OBJECTIVE: Our goal was to validate the Spanish version of the Brief Pain Inventory (BPI) questionnaire used to measure the intensity of oncological pain and its impact on activities of daily living in patients with cancer. PATIENTS AND METHOD: Patients with oncological pain were consecutively included in the study. These patients filled up the Spanish version of the BPI questionnaire (CBD) and the Rotterdam Symptom Checklist (RCSL) during the inclusion visit and again after 3-5 days (patients with clinically stable oncological pain) or after one month (patients with unstable oncological pain). RESULTS: 126 patients were assessed; 85.1% of them had suffered some episode of irruptive pain 24 hours prior to their inclusion in the study. 86.5% of patients fully completed the questionnaire. The CBD showed mild to moderate correlations with the patients perception of pain severity and with the presence of tumor dissemination. The <> dimension of the RCSL displayed the highest correlation with the dimensions of the BPI (<> and <>). The internal consistency and the test-retest reliability between dimensions were good (0.87 and 0.89) and low to moderate (0.53 and 0.77), respectively. The CBD questionnaire was found to be a tool capable of detecting changes in pain intensity. The changes observed in the two CBD dimensions between study visits fairly reflected the patients perceived changes in pain intensity. CONCLUSIONS: The Spanish version of BPI is valid for measuring the intensity of oncological pain and its impact on activities of daily living in conditions of usual clinical practice. PMID- 12570915 TI - [Body composition and health related quality of life as markers of efficacy of replacement therapy in adult patients with growth hormone deficiency]. PMID- 12570916 TI - [Pharmacogenetic studies: evaluation guidelines for research ethics committees. Scientific background and legal framework (I)]. PMID- 12570917 TI - [Advanced bladder cancer: new therapeutic strategies]. PMID- 12570918 TI - [Evolution of antibiotic consumption in Spain]. PMID- 12570919 TI - [Prolonged out-of-hospital prophylaxis of venous thromboembolism]. PMID- 12570920 TI - [Levofloxacin and bilateral spontaneous Achilles tendon rupture]. PMID- 12570921 TI - [RS3PE syndrome associated to AL-amyloidosis and multiple myeloma, with intestinal pseudo-obstruction]. PMID- 12570922 TI - Plasma leptin suppression by arginine vasopressin in normal women and men. AB - Leptin inhibits appetite by activating several neuroendocrine systems, including the hypothalamo-pituitary-adrenal cortical (HPA) axis. In turn, elevated glucocorticoids can increase circulating leptin. We therefore measured plasma leptin in 12 normal women and eight normal men administered low-dose physostigmine (PHYSO) and arginine vasopressin (AVP) to stimulate the HPA axis. The subjects underwent four test sessions 5-7 days apart: PHYSO (8 microg/kg IV), AVP (0.08 U/kg IM), PHYSO + AVP, and saline control. Serial blood samples were taken before and after pharmacologic challenge and analyzed for leptin, adrenocorticotropin (ACTH)1-39, cortisol, and AVP. Estradiol and testosterone also were measured at each test session. PHYSO and AVP produced no side effects in about half the subjects and predominantly mild side effects in the other half, with no significant female-male differences. Correlations between side effects (absent or present) after PHYSO or AVP and the corresponding leptin responses were nonsignificant. Baseline plasma leptin concentrations were significantly higher in the women than in the men (p < 0.003). Leptin concentrations following PHYSO remained unchanged from baseline, indicating that the short-lived ACTH and cortisol increases produced by PHYSO did not affect leptin secretion. In contrast, AVP administration, while also increasing ACTH and cortisol, suppressed leptin, to a significantly greater degree in the women than in the men (p = 0.01). This significant suppression of leptin by AVP has not been previously described; physiologically, it may be part of a negative feedback regulatory system between central leptin and its activation of the HPA axis, by inhibition of leptin production or acceleration of its clearance. PMID- 12570923 TI - Tramadol reduces the 5-HTP-induced head-twitch response in mice via the activation of mu and kappa opioid receptors. AB - Tramadol, an atypical opioid analgesic, stimulates both opiatergic and serotonergic systems. Here we have investigated the effect of tramadol in mice on 5-hydroxyptrytophan (5-HTP)-induced head twitch response (HTR), which is an animal model for the activation of the CNS 5-HT(2A) receptors in mice. Tramadol attenuated 5-HTP-induced HTR in a dose-dependent manner as morphine. Furthermore, the nonselective opioid receptor antagonists, naloxone and diprenorphine (M5050), reversed the effect of tramadol on 5-HTP-induced HTR dose-dependently. Interestingly, in contrast to the selective delta opioid receptor antagonist NTI, beta-FNA, a selective mu receptor antagonist, and nor-BNI, a selective kappa opioid receptor antagonist, antagonized the attenuation of 5-HTP-induced HTR by tramadol. In conclusion, administration of tramadol systemically inhibits 5-HTP induced HTR in mice by activating opiatergic system in the CNS. Our findings show that mu and kappa opioid receptors, but not delta opioid receptor, play an important role in the regulation of serotonergic function in the CNS. PMID- 12570924 TI - Pharmacokinetics of intravenous theophylline in mutant Nagase analbuminemic rats. AB - It was obtained from our laboratories that the expression of hepatic microsomal cytochrome P450 (CYP) 1A2 increased approximately 3.5 times in mutant Nagase analbuminemic rats (NARs, an animal model for human familial analbuminemia), and theophylline was reported to be metabolized to 1,3-dimethyluric acid (1,3-DMU) and 1-methylxanthine (which was further metabolized to 1-methyluric acid, 1-MU, via xanthine oxidase) via CYP1A2 in rats. Hence, the pharmacokinetic parameters of theophylline, 1,3-DMU and 1-MU were compared after intravenous administration of aminophylline, 5 mg/kg as theophylline, to control Sprague-Dawley rats and NARs. In NARs, the total area under the plasma concentration-time curve from time zero to time infinity (AUC) of theophylline was significantly smaller (1,040 versus 1,750 microg min/ml) than that in control rats and this could be due to significantly faster renal clearance (CL(R), 1.39 versus 0.571 ml/min/kg, due to inhibition of renal reabsorption of unchanged theophylline) and nonrenal clearance (CL(NR), 3.36 versus 2.25 ml/min/kg, due to 3.5-fold increase in CYP1A2) than those in control rats. Based on in vitro hepatic microsomal studies, the intrinsic 1,3-DMU formation clearance was significantly faster in NARs than that in control rats (267 versus 180 x 10(-6) ml/min). After intravenous administration of 1,3-DMU, the renal secretion of 1,3-DMU was inhibited in NARs. Inhibition of renal secretion or reabsorption of various compounds in NARs was also discussed. PMID- 12570925 TI - Magnesium-deficient medium enhances NO production in alveolar macrophages isolated from rats. AB - Magnesium deficiency has been shown to increase nitric oxide (NO) levels in plasma and to aggravate endotoxin lethality. The present study was performed to examine the effects of magnesium (Mg(2+))-deficient culture medium, with and without endotoxin (LPS), on NO release and inducible NOS (iNOS) mRNA levels in alveolar macrophages isolated from rats. Decreasing the Mg(2+) concentration in the culture medium from 0.39 mM (normal-Mg(2+) medium) to 0.021 mM (Mg(2+) deficient medium) increased NO release from alveolar macrophages for 2 h. However, LPS stimulation in Mg(2+)-deficient medium had little effect on NO release. The increased NO release in Mg(2+)-deficient medium was suppressed completely by L-NAME and aminoguanidine. Dexamethasone, pyrrolidine dithiocarbamate and curcumin strongly inhibited NO release. Verapamil, U73122, TMB-8 and W-7 had no significant effect on NO release induced by Mg(2+) deficiency. Preculture of macrophages with Mg(2+)-deficient medium for 22 h markedly increased NO release and iNOS mRNA levels for a further 2 h; these increments were suppressed completely by curcumin. These results suggest that Mg(2+) deficiency enhances NO production via iNOS by alveolar macrophages. In this experimental condition, we can not suggest that NO production from alveolar macrophage plays an essential role in the pathogenesis of enhanced endotoxin lethality in Mg-deficient rats. PMID- 12570926 TI - Functional role of ryanodine-sensitive Ca2+ stores in acidic pH-induced contraction in Wistar Kyoto rat aorta. AB - Acidic pH induced a contraction in the isolated aorta from Wistar Kyoto rat. The magnitude of contraction was dependent upon the degree of extracellular acidification. The maximum level of contraction observed at pH 6.5 was 84.6 +/- 3.4% of the 64.8 mM KCl-induced contraction. To investigate the role of extracellular as well as intracellular Ca(2+) in acidic pH-induced contraction (APIC), we changed the extracellular pH in the presence of EGTA. Sustained contraction induced by acidic pH in the presence of extracellular Ca(2+) was completely abolished in the presence of EGTA, while a transient but significant contraction was still observed. Ryanodine, a selective ryanodine receptor blocker and cyclopiazonic acid (CPA), an inhibitor of sarco-/endoplasmic reticulum Ca(2+) ATPase, abolished the transient contraction, when pH was decreased in Ca(2+)-free solution. On the other hand, neither xestospongin C, a selective inositol-1,4,5 trisphosphate receptor antagonist nor U-73122, a phospholipase C inhibitor showed this effect. These results suggest the involvement of Ca(2+) release from ryanodine-/CPA-sensitive store of sarcoplasmic reticulum (SR). In normal Ca(2+) containing solution, ryanodine and CPA did not alter the maximum level of APIC. However, they significantly decreased the rate of rise of APIC. U-73122, suppressed the maximum contraction induced by acidic pH without affecting the rate of rise of APIC, while xestospongin C and U-73343, an inactive analogue of U 73122, had no effect on both parameters of APIC. From these results, it is concluded that acidic pH induces Ca(2+) release from the ryanodine-/CPA-sensitive store of SR and that release provides supportive effect on initiating rapid transient contraction, but not on the sustained contraction, which is entirely due to Ca(2+) influx. PMID- 12570927 TI - Fullerene derivative attenuates ischemia-reperfusion-induced lung injury. AB - Reactive oxygen species are the major contributing factors to lung ischemia reperfusion (IR) injury. In this study, we tested whether a water soluble antioxidant fullerene derivative [C(60)(ONO(2))(7 +/- 2)] attenuates IR lung injury. Young Wistar rats were divided into two groups: control and C(60)(ONO(2))(7 +/- 2). Under ventilation with 95% air-5% CO(2) gas mixture and a 2.5 cm H(2)O end-expiratory pressure, the isolated lungs were perfused with a physiological solution. The experimental protocol included three periods: baseline (10 min), ischemia (45 min) and reperfusion (60 min, ventilated with 95% O(2)-5% CO(2) gas mixture). Before and after ischemia, we measured pulmonary arterial pressure (Ppa), pulmonary venous pressure and lung weight (W). Then, pulmonary capillary pressure and filtration coefficient (K(fc)) were calculated. Ischemia caused increases in Ppa, W and K(fc) in the control group. For most cases, the above ischemia-induced increases were attenuated by the C(60)(ONO(2))(7 +/- 2) pretreatment. Our results suggest that the antioxidant C(60)(ONO(2))(7 +/- 2) attenuates IR-induced lung injury. PMID- 12570928 TI - Hormone replacement therapy increases renal kallikrein excretion in healthy postmenopausal women. AB - Hypertension and its related increase in cardiovascular morbidity in postmenopausal women is a major public health problem. The hypotensive property of urinary kallikrein has been described since 1909. Despite the controversy surrounding the effects of hormone replacement therapy on blood pressure regulation, its mechanisms remain incompletely understood, and no evidence has yet been provided for its effects on renal kallikrein excretion in postmenopausal women. In a double-blind, randomized study we examined the effects of hormone replacement therapy in the form of 2 mg 17-beta estradiol (ERT) or 2 mg 17-beta estradiol combined with continuous 5 mg medroxyprogesterone acetate (HRT) on urinary kallikrein excretion in postmenopausal women. Thirty-nine postmenopausal women collected their urine for 24 hours on two separate occasions 3 months apart. During the 3 month period women were randomized to placebo, ERT, or HRT. Urine samples were assayed for kallikrein activity, normalized to urine creatinine and expressed as mU/gm creatinine. Urinary kallikrein excretion increased significantly after 3 months in the ERT (p < 0.001) and HRT (p < 0.01) groups, and decreased non-significantly in the placebo group (p > 0.06). There were no significant blood pressure changes after 3 months of therapy. The findings demonstrate that hormone replacement therapy in the form of estrogen or estrogen combined with continuous medroxyprogesterone is effective in increasing urinary kallikrein excretion. Given that a decrease in kallikrein excretion may mark risk for development of hypertension, the findings of this study are of value in demonstrating a novel mechanism underlying cardioprotective properties of postmenopausal hormone replacement therapy in women without pre-existing coronary disease. PMID- 12570929 TI - Intercellular communication between renin expressing As4.1 cells, endothelial cells and smooth muscle cells. AB - Angiotensin II (AII) regulation of renin production by the juxtaglomerular (JG) cells of the kidney is commonly thought to occur through a direct feedback mechanism. However, recent evidence suggests that other cells in the vicinity may indirectly mediate AII's effect on renin production. Therefore we investigated whether an in vitro model of JG cells (As4.1) could have intercellular communication with endothelial or smooth muscle cells, which are in proximity to JG cells in vivo. 6-carboxyfluorescein was introduced to individual bovine aortic endothelial cells in co-culture with As4.1 cells. Coupling was observed 84% of the time at resting membrane potential and was attenuated by membrane depolarization or octanol (1 mM). Calcein green transfer between human aortic smooth muscle and As4.1 cells occurred 82% of the time and was inhibited by octanol. Expression of connexin 37, 40, 43, and 45 were detected in As4.1 cells using RT-PCR. Stimulation of As4.1 cells by AII failed to alter [Ca(2+)](i) or renin mRNA levels. These findings support the existence of gap junctions between renin producing cells and other cell types of the JG region. Moreover the lack of effect by AII suggest that feedback regulation of renin by AII may be due in part to intercellular communication with cells in proximity to JG cells. PMID- 12570930 TI - Effects of cadmium on E-cadherin and VE-cadherin in mouse lung. AB - Exposure to Cd(2+) via inhalation or intratracheal instillation results in pulmonary edema, which is followed by the influx of leukocytes, the proliferation of type II pneumocytes and eventual scarring and fibrotic changes. While the general toxic effects of Cd(2+) in the lung have been well characterized, the specific molecular mechanisms underlying these effects have yet to be elucidated. Previously we have shown that Cd(2+) can disrupt the adhering junctions between various types of epithelial and endothelial cells in culture, most likely by perturbing the function of the Ca(2+) dependent cell adhesion molecules E cadherin and VE-cadherin respectively. The objectives of this study were to determine whether respiratory exposure to Cd(2+) can alter the localization of E cadherin and VE-cadherin in the lung, and to determine whether this effect may play a role in the acute pneumotoxic response to Cd(2+). Male CF-1 mice were exposed to CdCl(2) (0, 16.25, 32.5, 65 or 130 nmoles in 50 microl saline) via intratracheal instillation. After 24 hours, the lungs were removed and either subjected to bronchoalveolar lavage or analyzed for histopathologic changes. The results showed that Cd(2+) caused an increase in lung weight and in the protein content of the lavage fluid. These effects were accompanied by a pronounced decrease in the amount of E-cadherin in epithelial cells of the alveoli and small bronchioles and of VE-cadherin in vascular endothelial cells. Assessment of cell membrane integrity with ethidium homodimer-1 showed no evidence of severe injury or death in alveolar epithelial cells. These findings suggest that E-cadherin and VE-cadherin may be important early targets of Cd(2+) toxicity in the lung. PMID- 12570931 TI - Modulation of visual event-related potentials by emotional olfactory stimuli. AB - The objective of the present study was to determine whether an olfactory prime could modulate behavior and visual event-related potentials (ERPs) obtained in response to a visual stimulation representing female faces. More specifically, we tested the hypothesis that a pleasant odor could have effects on face perception: behavioral effects on subjective emotional estimation of faces, and on associated response times, and electrophysiological effects on the N400 and late positive complex or LPC. Experiments were performed in which subjects had to decide whether the presented face was pleasant or not, while visual ERPs were recorded. Faces were always primed with either a pleasant odor or a neutral olfactory stimulus (pure air). In order to test the effect of subject's awareness, participants were not informed that an odor would be presented in the experimental sessions. Responses were significantly shorter for unpleasant faces. However, no behavioral effects of the pleasant odor on response time or on evaluation of face pleasantness were observed. Late ERPs evoked by faces were modulated by the presence of a pleasant odor, even when subjects were neither warned nor aware of the presence of the odor: in a frontal site and after the diffusion of the odor, the LPC (appearing 550 ms after the presentation of the visual stimulus) evoked by unpleasant faces was significantly more positive than the LPC evoked by pleasant faces. This effect could reflect an enhanced alert reaction to unpleasant faces are preceded by an (incongrous) pleasant odor. PMID- 12570932 TI - Study of a patient population investigated for excessive daytime sleepiness (EDS). AB - This study included all patients referred to the out-patient department of our sleep disorders centre from 1993 to 1999 on account of excessive daytime sleepiness (EDS). As a first step, patients in whom a diagnosis was established following appropriate polysomnography were excluded: this included sleep apnea syndrome, increased upper airway resistance syndrome, narcolepsy, periodic movements during sleep or other parasomnia, and epilepsy. Patients regularly taking psychotropic substances or with psychiatric disorders were also excluded. Finally, 128 patients remained in whom no clear diagnosis had been established for EDS, 70 women and 58 men, their ages ranging from 16 to 77 years. They underwent a 48-h recording (night 1-MSLT-night 2-continuous day). The aim of the study was to establish, define and characterise different groups of undiagnosed EDS patients using clinical, electrophysiological and immunological data with the help of hierarchical cluster analysis. Eight groups were characterised: group 1: mild hypersomnia type 1 (n = 11); group 2: hypersomnia frequently associated with HLA type DR2-DQw1 (n = 11); group 3: mild hypersomnia type 2 (n = 28); group 4: morning recovery from disrupted sleep (n = 19); group 5: young "long sleepers with difficulty at waking up" (n = 17); group 6: idiopathic hypersomnia (n = 15); group 7: poor or short sleepers since childhood (n = 8); group 8: older poor sleepers with a late onset of symptoms (n = 19). Characteristic features of these different groups provided consistent and objective arguments leading to a more precise diagnosis for these patients, and helped the initiation of appropriate management and treatment. PMID- 12570933 TI - [Anticipatory postural adjustments associated with arm flexion: interest of vertical torque]. AB - OBJECTIVE: To find a biomechanical parameter able to characterize postural adjustments in different movement conditions. METHODS: The arm-raising movement performed during the upright human position imposes a vertical torque (Tz) that can be measured by a force plate-form. This torque was studied in ten healthy young subjects with opto electronic system Vicon 370. The subjects stood on a force platform, performed shoulder flexion of their right arm, to grasp a handle in front of them, in five conditions : self-paced at 3 different velocities (slow, medium, maximal), triggered by an auditory signal, loaded (1 kg attached to the wrist), all at maximal velocity. In a sixth condition, the arm was passively displaced by an experimenter. RESULTS: Tz displayed a negative phase (counter-clock wise body rotation) in all conditions. A positive phase occurred prior to the negative one, and preceded movement onset only if the movement was voluntary. In the triggered condition, the positive phase of Tz was delayed (- 60 ms) compared to the self-paced condition at maximal velocity (- 155 ms). However Tz onset latency was modified neither by load nor velocity. Tz amplitude increased with increasing velocity, load and in a reaction time condition. CONCLUSIONS: The vertical torque Tz, especially its positive phase gives useful informations about the latency, duration and intensity of the postural preparation related to a voluntary movement, according to the movement parameters. Tz is therefore able to characterise postural adjustments in all conditions, even with low movement velocity. PMID- 12570934 TI - [Spatial orienting of attention: a study of reaction time during pointing movement]. AB - AIM: The aim of this study was to investigate how advance information both explicit and implicit provided prior to movement may affect the spatial orientation and the internal attention control processes in normal adult subjects. The originality of this work compared to the test of Posner, lies essentially in the methodology used to study the attentional systems. The use of three procedures of reaction time (RT) allowed us to study the setting concerned of the specific and non-specific components of the attention in the motor preparation. By associating of these three procedures of RT, we have evaluated the effects of the explicit and implicit components of advance information on motor preparation. The use of advance information to the movement requires the implication of the attentional systems. MATERIAL AND METHODS: Experiments were carried out using a simple reaction time (RT) procedure involving the use of an orientation cue and two choice reaction time situations: one with a neutral preparatory cue and one with a priming cue giving the likelihood of the preparatory stimulus (S1) being compatible with the imperative stimulus (S2). The mechanisms underlying the subjects' vigilance and the orientation of their attention were studied by assessing the effects on their reaction times of the preparatory signal and those of the cue giving the likelihood of S1 and S2 being compatible. The preparatory signal was designed to explicitly attract the subjects' attention towards the position of the forthcoming pointing target, whereas the cue giving the compatibility between S1 and S2 was intended to mobilize the subject's attention more implicitly. Prior to performing the pointing movement towards a visual target, the subjects' attention was therefore mobilized by the advance information containing two components: the explicit information about the position of S1 and the implicit information about the probability of S1 and S2 being compatible. RESULTS AND CONCLUSION: The results obtained here on 17 normal adult subjects show that the subjects significantly improved their RTs by using the explicit component of the information provided. The implicit information available was also used in the choice reaction situations: a priming effect was found to occur, which resulted in the shortening of the primed "compatible cue" reaction times in comparison with the "neutral cue" reaction times, and in the correlation which was found to exist between the reaction time performances and the degree of compatibility between the preparatory signal and the imperative signal. These results suggest that various components of the attentional system may participate in processing the advance information provided prior to the movement in reaction time tasks of the kind used here. The explicit information provided prior to the movement may mobilize the subject's vigilance and spatially orients his attention; whereas the implicit information available may rather subserve the internal control of the subject's attention. PMID- 12570936 TI - Association between hyperglycemia and the no-reflow phenomenon in patients with acute myocardial infarction. AB - OBJECTIVES: We investigated the association between hyperglycemia and the no reflow phenomenon in patients with acute myocardial infarction (AMI). BACKGROUND: Hyperglycemia is associated with increased risks of heart failure, cardiogenic shock, and death after AMI, but its underlying mechanism remains unknown. METHODS: A total of 146 consecutive patients with a first AMI were studied by intracoronary myocardial contrast echocardiography (MCE) after successful reperfusion within 24 h after symptom onset. Two-dimensional echocardiography was recorded on day 1 and three months later to determine the change in the wall motion score (DeltaWMS; sum of 16 segmental scores; dyskinesia = 4 to normokinesia = 0). RESULTS: The no-reflow phenomenon was found on MCE in 49 (33.6%) of 146 patients; their glucose level on hospital admission was significantly higher than that of patients who did not exhibit this phenomenon (209 +/- 79 vs. 159 +/- 56 mg/dl; p < 0.0001). There was no difference in glycosylated hemoglobin or in the incidence of diabetes mellitus between the two subsets. The no-reflow phenomenon was more often observed in the 75 patients with hyperglycemia (>/=160 mg/dl) than in those without hyperglycemia (52.0% vs. 14.1%; p < 0.0001). Patients with hyperglycemia had a higher peak creatine kinase level (2,497 +/- 1,603 vs. 1,804 +/- 1,300 IU/l; p = 0.005) and a lower DeltaWMS (3.7 +/- 4.8 vs. 5.7 +/- 4.3; p = 0.01) than did those without hyperglycemia. The blood glucose level was an independent prognostic factor for no reflow, along with age, gender, absence of pre-infarction angina, complete occlusion of the culprit lesion, and anterior AMI. CONCLUSIONS: Hyperglycemia might be associated with impaired microvascular function after AMI, resulting in a larger infarct size and worse functional recovery. PMID- 12570937 TI - Enoxaparin in unstable angina patients who would have been excluded from randomized pivotal trials. AB - OBJECTIVES: In the present study, we describe the characteristics and examine the anticoagulation levels and safety of subcutaneous enoxaparin in unstable angina (UA)/non-ST-segment elevation myocardial infarction (NSTEMI) patients who would not have been eligible in the Efficacy Safety Subcutaneous Enoxaparin in Non-Q wave Coronary Events (ESSENCE) and Thrombolysis In Myocardial Infarction (TIMI) 11B trials. BACKGROUND: It is not known whether the benefit shown with enoxaparin in the selected population of pivotal trials can be extended to the real world. METHODS: In our center, all patients with UA/NSTEMI are anticoagulated with subcutaneous enoxaparin adjusted to creatinine clearance. Among 515 consecutive patients, we identified 174 who would not have been eligible for ESSENCE or TIMI 11B ("EP" group for excluded patients). We evaluated cardiovascular death or non fatal myocardial infarction (MI), as well as major and minor bleeding events, at 30 days in the EP group and in patients without any of the exclusion criteria ("NEP" group for non-excluded patients). RESULTS: This EP group was older, had a higher female/male ratio, and more frequently had a history of MI or a diagnosis of non-Q MI on admission than the NEP group. The distribution of the anti-Xa activity was similar in both groups. The bleeding rates (major and minor) at 30 days were similar in the EP and NEP groups (2.3% vs. 2.9%, respectively, P = NS). On multivariate analysis, the use of glycoprotein IIb/IIIa inhibitors and the presence of hypertension were the only independent predictors of bleeding found in the whole population. Compared with the NEP group, the EP group had a fourfold increased rate of death or MI at 30 days (15.5% vs. 4.1%, p < 0.01). On multivariate analysis, the independent predictors of death or MI at 30 days were NSTEMI on admission, creatinine clearance, and heart failure. CONCLUSIONS: Patients who do not fit the enrollment criteria of ESSENCE/TIMI-11B have higher risk baseline characteristics for both bleeding and ischemic events. In these patients, enoxaparin with dose adjustment to creatinine clearance provides adequate anti-Xa levels and no excess of bleeding. PMID- 12570938 TI - Rho-kinase inhibition with intracoronary fasudil prevents myocardial ischemia in patients with coronary microvascular spasm. AB - OBJECTIVES: We sought to determine whether a potent Rho-kinase inhibitor fasudil prevents the occurrence of myocardial ischemia in patients with microvascular angina attributable to coronary microvascular spasm. BACKGROUND: Effective treatment of patients with angina who have normal coronary arteriograms (microvascular angina) has not yet been established. Rho-kinase-mediated calcium sensitization of the myosin light chain in smooth muscle cells has been implicated as substantially contributing to vascular hyperconstriction. METHODS: We studied consecutive 18 patients with angina and normal epicardial coronaries in whom intracoronary acetylcholine (ACh) induced myocardial ischemia (ischemic electrocardiographic changes, myocardial lactate production, or both) without angiographically demonstrable epicardial coronary vasospasm. All patients underwent a second ACh challenge test after pretreatment with either saline (n = 5) or fasudil (4.5 mg intracoronarily, n = 13). RESULTS: Myocardial ischemia was reproducibly induced by ACh in the saline group. In contrast, 11 of the 13 patients pretreated with fasudil had no evidence of myocardial ischemia during the second infusion of ACh (p < 0.01). The lactate extraction ratio (median value [interquartile range]) during ACh infusion was improved by fasudil pretreatment, from -0.16 (-0.25 to 0.04) to 0.09 (0.05 to 0.18) (p = 0.0125). CONCLUSIONS: Fasudil ameliorated myocardial ischemia in patients who were most likely having coronary microvascular spasm. The inhibition of Rho-kinase may be a novel therapeutic strategy for this group of patients with microvascular angina. PMID- 12570939 TI - Safety of concomitant therapy with eptifibatide and enoxaparin in patients undergoing percutaneous coronary intervention: results of the Coronary Revascularization Using Integrilin and Single bolus Enoxaparin Study. AB - OBJECTIVES: This study was designed to assess whether use of enoxaparin during percutaneous coronary intervention (PCI) increased bleeding compared with unfractionated heparin, in addition to background therapy with eptifibatide. BACKGROUND: Data supporting the benefits of enoxaparin and the glycoprotein IIb/IIIa inhibitor eptifibatide evolved in parallel. Information on combining these two classes of medications is limited. METHODS: A total of 261 patients undergoing elective or urgent PCI were randomized to either eptifibatide plus enoxaparin or eptifibatide plus unfractionated heparin. RESULTS: The primary end point of the study, the bleeding index (change in hemoglobin corrected for blood transfusions), was 0.8 in the patients randomized to enoxaparin and 1.1 in patients randomized to unfractionated heparin (p = 0.15). The rate of vascular access site complications was 9.3% in the enoxaparin arm versus 9.8% in the unfractionated heparin arm (p = NS). The rate of bleeding complications was not significantly different between the two arms of the study, including in those patients who received vascular closure devices. The rate of angiographic complications was 6.3% in the enoxaparin group and 6.2% in the unfractionated heparin group (p = NS). Similarly, there were no significant differences in the composite of death, myocardial infarction, or urgent target vessel revascularization at 48 h or 30 days. CONCLUSIONS: Compared with unfractionated heparin plus eptifibatide, the combination of enoxaparin plus eptifibatide is not associated with an excess of bleeding or vascular complications, including in those receiving closure devices. Despite no monitoring of anticoagulation activity with enoxaparin, there was no apparent increase in angiographic or clinical complications. PMID- 12570940 TI - Intravenous glycoprotein IIb/IIIa receptor antagonists reduce mortality after percutaneous coronary interventions. AB - OBJECTIVES: We sought to evaluate the impact of intravenous antagonists of the platelet IIb/IIIa receptor on the survival of patients undergoing percutaneous coronary interventions (PCIs). BACKGROUND: Several trials have shown that intravenous antagonists of the platelet glycoprotein (GP) IIb/IIIa receptor reduce the incidence of myocardial infarction (MI) and composite cardiac outcomes (death, MI, or revascularization) in patients undergoing PCI. However, individual studies have not had adequate power to examine differences in mortality. METHODS: We performed a meta-analysis of 19 randomized, placebo-controlled trials (20 comparisons, n = 20,137). Death was the primary outcome. Secondary outcomes included MI, composite cardiac outcomes, and major bleeding. RESULTS: Mortality was significantly reduced at 30 days (risk ratio [RR] 0.69 [95% confidence interval [CI] 0.53 to 0.90]), at six months (RR 0.79 [95% CI 0.64 to 0.97]), and including longer follow-up (RR 0.79 [95% CI 0.66 to 0.94]), with no significant between-study heterogeneity. The relative risk reduction was largely similar in trials of patients with or without acute myocardial infarction (AMI), in trials continuing or discontinuing heparin after the procedure, and in trials using stents or another PCI as the intended primary procedure. Myocardial infarction and composite outcomes were significantly reduced (p < 0.001 for all) at 30 days and six months. Major bleeding was significantly increased only in trials where heparin infusion was continued after the procedure (RR 1.70 [95% CI 1.36 to 2.14]), although there was no excess bleeding when heparin was discontinued (RR 1.02 [95% CI 0.85 to 1.24]). CONCLUSIONS: In patients undergoing PCI, GP IIb/IIIa receptor antagonists confer a significant and sustained decrease (20% to 30%) in the risk of death. PMID- 12570941 TI - Postprocedure chest pain after coronary stenting: implications on clinical restenosis. AB - OBJECTIVES: The goal of this study was to analyze the incidence and predictors of postprocedure chest pain (PPCP) after percutaneous coronary intervention (PCI) and its correlation with clinical restenosis. BACKGROUND: Chest pain after PCI occurs frequently even in the absence of procedural events and is considered to be due to vasospasm or coronary artery stretch. The short- and long-term significance of PPCP after otherwise successful stenting is not clear. METHODS: We analyzed 1,362 patients undergoing coronary stenting for PPCP, procedural and in-hospital events, 30-day major adverse cardiac events, and target vessel revascularization (TVR) at 6 to 9 months. RESULTS: There were 488 patients with PPCP and, of these, 312 patients were excluded due to procedural events. The remaining 176 patients with PPCP were compared with 874 patients without PPCP. Creatine kinase-MB isoenzyme elevation occurred in 25.6% of the PPCP group versus 9.6% of the no PPCP group (p < 0.001). Despite similar reference vessel diameter, the PPCP group had larger postprocedure minimum lumen diameter, higher stent-to vessel ratio, and higher inflation pressure versus the no PPCP group (p < 0.01). At 30 days, the emergency room visits and repeat catheterization (16% vs. 2.7%; p < 0.001) were higher in the PPCP group versus the no PPCP group, but repeat intervention was similar. At 6- to 9-month follow-up, the TVR was significantly higher in the PPCP group compared with the no PPCP group (29.5% vs. 16.6%; p < 0.01). CONCLUSIONS: Our analysis suggests micromyonecrosis and vessel stretch as causes of PPCP. Postprocedure chest pain is associated with similar short-term outcome as no PPCP, but has higher restenosis, perhaps mediated by deep vessel wall injury. Therefore, PPCP may identify patients at high risk for restenosis. PMID- 12570942 TI - The prevalence and severity of coronary artery calcification on coronary artery computed tomography in black and white subjects. AB - OBJECTIVES: We studied the relationship between coronary artery calcium (CAC) and race in asymptomatic, active-duty personnel in the Prospective Army Coronary Calcium (PACC) project. BACKGROUND: Valid cardiovascular risk assessments in black Americans using coronary artery computed tomography (coronary CT) require the generalizability of population-based CAC score distributions derived from primarily white patient populations. METHODS: Among 1,000 consecutive participants (mean age, 42 +/- 2 years; range, 40 to 45 years), 999 participants underwent coronary CT and indicated a specific racial affiliation. This included white, non-Hispanic in 699 (69.9%) participants and black, non-Hispanic in 194 (19.4%) participants. Univariate associations between race and cardiovascular risk variables were entered into a logistic regression model for CAC that also controlled for socioeconomic status and education. RESULTS: Coronary artery calcium was nearly twice as prevalent in white (19.2%) than in black participants (10.3%) (p = 0.004). Black individuals had a threefold greater prevalence of hypertension, left ventricular hypertrophy, ST-T-wave abnormalities, and current cigarette smoking. Black subjects also had significantly greater blood pressure, high-density lipoprotein cholesterol, glycosylated hemoglobin, lipoprotein(a) and fibrinogen levels, and lower triglyceride levels and waist girth than white subjects. After adjustment for these differences, and socioeconomic adjusters, black individuals were 39% as likely to have any CAC present (odds ratio, 0.39; 95% confidence interval, 0.20 to 0.78; p = 0.007). CONCLUSIONS: Despite a worse cardiovascular risk profile, black Americans have significantly less CAC than white Americans. The use of coronary CT as an accurate risk prediction tool in black Americans will require ethnic-specific data on the presence and severity of CAC. PMID- 12570943 TI - The ethnic-specific nature of mechanisms for coronary heart disease. PMID- 12570944 TI - Level of kidney function as a risk factor for atherosclerotic cardiovascular outcomes in the community. AB - OBJECTIVES: The goal of this study was to determine whether the level of kidney function is an independent risk factor for atherosclerotic cardiovascular disease (ASCVD) outcomes in the Atherosclerosis Risk in Communities (ARIC) study, a prospective cohort study of subjects aged 45 to 64 years. BACKGROUND: The level of kidney function is now recognized as a risk factor for ASCVD outcomes in patients at high risk for ASCVD, but it remains unknown whether the level of kidney function is a risk factor for ASCVD outcomes in the community. METHODS: Cox proportional-hazards regression was used to evaluate the association of glomerular filtration rate (GFR) with ASCVD after adjustment for the major ASCVD risk factors in 15,350 subjects. We searched for nonlinear relationships between GFR and ASCVD. RESULTS: During a mean follow-up time of 6.2 years, 965 (6.3%) of subjects had ASCVD events. Subjects with GFR of 15 to 59 ml/min/1.73 m(2) (n = 444, hazard ratio 1.38 [1.02, 1.87]) and 60 to 89 ml/min/1.73 m(2) (n = 7,665, hazard ratio 1.16 [1.00, 1.34]) had an increased adjusted risk of ASCVD compared with subjects with GFR of 90 to 150 ml/min/1.73 m(2). Each 10 ml/min/1.73 m(2) lower GFR was associated with an adjusted hazard ratio of 1.05 (1.02, 1.09), 1.07 (1.01, 1.12), and 1.06 (0.99, 1.13) for ASCVD, de novo ASCVD, and recurrent ASCVD, respectively. A nonlinear model did not fit the data better than a linear model. CONCLUSIONS: The level of GFR is an independent risk factor for ASCVD and de novo ASCVD in the ARIC study. PMID- 12570945 TI - The underutilization of cardiac medications of proven benefit, 1990 to 2002. AB - OBJECTIVES: To evaluate recent trends, we examined longitudinal national data on the outpatient use of warfarin in atrial fibrillation (AF), beta-blockers and aspirin in coronary artery disease (CAD), and angiotensin-converting enzyme inhibitors (ACEIs) in congestive heart failure (CHF). BACKGROUND: Previous studies indicate that specific cardiac medications are underutilized. METHODS: We used the National Disease and Therapeutic Index (NDTI) (produced by IMS HEALTH, Plymouth Meeting, Pennsylvania) for 1990 to 2002, and the National Ambulatory Medical Care Surveys (NAMCS) for 1990 to 2000 to follow nationally representative samples of outpatient visits. For visits by patients with AF (total n = 14,634 visits), CAD (n = 35,295), and CHF (n = 33,008), we examined trends in the proportion of visits with the selected medications reported. RESULTS: Warfarin use in AF increased from 12% in 1990, to 41% in 1995, to 58% in 2001 in NDTI; a similar moderation of recent increase was seen in NAMCS. For CAD in NDTI, beta blocker use increased slowly from 19% in 1990, to 20% in 1995, then to 40% in 2001; NAMCS showed this same pattern. Aspirin use in CAD in NDTI increased from 18% in 1990, to 19% in 1995, to 38% in 2001; NAMCS, however, showed lower use rates. For NDTI, ACEI use in CHF increased from 24% in 1990 to 36% in 1996, but increased to only 39% by 2001, a general pattern also seen in NAMCS. CONCLUSIONS: Both national datasets demonstrate continuing underutilization of these cardiac medications of proven benefit. Although use is increasing, it remains lower than expected, and some increases noted in earlier years have slowed. Substantial public health benefits would result from further adoption of these effective therapies. PMID- 12570946 TI - Cardiology participation improves outcomes in patients with new-onset heart failure in the outpatient setting. AB - OBJECTIVES: This study examined the outcomes of new-onset heart failure (HF) outpatients managed by cardiologists and primary care (PC) physicians. BACKGROUND: Several studies have sought differences in outcomes between patients with HF managed by cardiologists and PC physicians, but most focused on inpatients, who often represent later stages of HF, whereas many treatments have their impact by delaying disease progression. METHODS: This was a retrospective cohort study of incident HF identified between 1996 and 1997 in a staff model health maintenance organization. Cardiology care was defined as >/=2 visits or >/=25% of total medical outpatient visits to cardiology. Records from a cohort of 403 patients with new-onset outpatient HF were reviewed. The main outcome measure was a combination of death and/or cardiovascular hospitalization at 24 months. RESULTS: Cardiologists' patients (n = 198) were younger (66 vs. 71 years, p = 0.001), were more likely men (54% vs. 46%, p = 0.01), had coronary artery disease (64% vs. 42%, p = 0.001), and had a low (/=50 ms. RESULTS: One area of E-IDC was recorded in 20 patients, and 2 or more were recorded in 4 patients. In 23 patients, these areas were detected during RVA pacing; in only 14 during SR. An E-IDC area related to the clinical VT was identified in each patient. Ablation guided by E-IDC suppressed all but one clinical VT whose inducibility suppression was tested. During a follow-up period of 9 +/- 4 months, three patients had recurrences of the ablated VT and two of a different VT. CONCLUSIONS: Electrograms with IDCs related to clinical VT can be identified in the majority of patients during RVA pacing. Radiofrequency ablation of E-IDC seems effective in controlling unmappable VT. PMID- 12570950 TI - Mechanism of syncope in patients with positive adenosine triphosphate tests. AB - OBJECTIVES: We prospectively evaluated the mechanism of syncope in patients with positive adenosine triphosphate (ATP) tests (defined as the induction of atrioventricular [AV] block with a ventricular pause >/=6 s after an intravenous bolus of 20 mg ATP). BACKGROUND: Patients with unexplained syncope tend to have more positive ATP tests results than those without syncope. METHODS: An implantable loop recorder (ILR) was inserted in 36 ATP-positive patients (69 +/- 10 years; 22 women; median of 6 syncopal episodes); 15 of them also had a positive response to tilt testing. RESULTS: During the follow-up of 18 +/- 9 months, 18 patients (50%) had syncopal recurrence and 16 (44%) had an electrocardiographically documented episode: AV block (n = 3: paroxysmal in 2 and permanent in 1), AV block followed by sinus arrest (n = 1), sinus arrest (n = 5), sinus bradycardia <40 beats/min (n = 2), normal sinus rhythm (n = 2), sinus tachycardia (n = 1), rapid atrial fibrillation (n = 1), and ectopic atrial tachycardia (n = 1). Bradycardia was documented in a total of 11 cases (69%), and a long ventricular pause (4 to 29 s) was present in eight cases (50%). All three patients with ILR-documented AV block had previously had a negative tilt test, whereas seven of eight with ILR-documented sinus bradycardia or sinus arrest had previously had a positive tilt test. CONCLUSIONS: In patients with adenosine sensitive syncope, the mechanism of syncope is heterogeneous, although bradycardia is the most frequent finding. Adenosine triphosphate-induced AV block predicts AV block as the mechanism of spontaneous syncope in only a few tilt negative patients. PMID- 12570951 TI - Upper and lower limits of vulnerability to sudden arrhythmic death with chest wall impact (commotio cordis). AB - OBJECTIVES: In an animal model of commotio cordis, sudden death with chest-wall impact, we sought to systematically evaluate the importance of impact velocity in the generation of ventricular fibrillation (VF) with baseball chest-wall impact. BACKGROUND: Sudden cardiac death can occur with chest-wall blows in recreational and competitive sports (commotio cordis). Analyses of clinical events suggest that the energy of impact is often not of unusual force, although this has been difficult to quantify. METHODS: Juvenile swine (8 to 25 kg) were anesthetized, placed prone in a sling to receive chest-wall strikes during the vulnerable time window during repolarization for initiation of VF with a baseball propelled at 20 to 70 mph. RESULTS: Impacts at 20 mph did not induce VF; incidence of VF increased incrementally from 7% with 25 mph impacts, to 68% with chest impact at 40 mph, and then diminished at >/=50 mph (p < 0.0001). Peak left ventricular pressure generated by the chest blow was related to the incidence of VF in a similar Gaussian relationship (p < 0.0001). CONCLUSIONS: The energy of impact is an important variable in the generation of VF with chest-wall impacts. Impacts at 40 mph were more likely to produce VF than impacts with greater or lesser velocities, suggesting that the predilection for commotio cordis is related in a complex manner to the precise velocity of chest-wall impact. PMID- 12570952 TI - Low vitamin D status: a contributing factor in the pathogenesis of congestive heart failure? AB - OBJECTIVES: This study was designed to evaluate the association between vitamin D status and congestive heart failure (CHF). BACKGROUND: Impaired intracellular calcium metabolism is an important factor in the pathogenesis of CHF. The etiology of CHF, however, is not well understood. METHODS: Twenty patients age <50 years and 34 patients age >/=50 years with New York Heart Association classes >/=2 and 34 control subjects age >/=50 years were recruited. N-terminal pro atrial natriuretic peptide (NT-proANP), a predictor of CHF severity; vitamin D metabolites; and parameters of calcium metabolism were measured in fasting blood samples collected between November 2000 and March 2001. RESULTS: Both groups of CHF patients had markedly increased serum levels of NT-proANP (p < 0.001), increased serum phosphorus levels (p < 0.001), and reduced circulating levels of both 25-hydroxyvitamin D (p < 0.001) and calcitriol (p < 0.001). Albumin corrected calcium levels were reduced and parathyroid hormone levels were increased in the younger CHF patients compared with the controls (both p values <0.001). Moreover, parathyroid hormone levels tended to be higher in the elderly CHF patients than in the controls (p = 0.074). In a nonlinear regression analysis 25-hydroxyvitamin D and calcitriol were inversely correlated with NT-proANP (r(2) = 0.16; p < 0.001 and r(2) = 0.12; p < 0.01, respectively). The vitamin D genotype at the BmsI restriction site did not differ between the study groups. CONCLUSIONS: The low vitamin D status can explain alterations in mineral metabolism as well as myocardial dysfunction in the CHF patients, and it may therefore be a contributing factor in the pathogenesis of CHF. PMID- 12570953 TI - Retrospective analysis of the cost-effectiveness of using plasma brain natriuretic peptide in screening for left ventricular systolic dysfunction in the general population. AB - OBJECTIVES: We sought to assess the cost-effectiveness of using plasma brain natriuretic peptide (BNP) as a pre-echocardiographic screening test for left ventricular systolic dysfunction (LVSD) in the general population. BACKGROUND: We hypothesized that plasma BNP and simple clinical parameters would reduce the number of echocardiograms needed and cost when screening for LVSD in the general population. METHODS: A random sample of 1,257 community subjects (age 25 to 74 years) was examined. Three risk groups were formed: one group with symptomatic ischemic heart disease (IHD); a second group with blood pressure >160/95 mm Hg and/or an abnormal electrocardiogram (high risk); and a group with none of these risk factors (low risk). The BNP assay was adjusted to give a high sensitivity. RESULTS: Left ventricular systolic dysfunction was prevalent in 0.7% (6/823), 6% (16/269), and 19% (26/140) of low-risk and high-risk subjects and IHD subjects, respectively. Raised BNP concentrations (>8 pg/ml) occurred in 41%, 64%, and 71%. Sensitivities of BNP for detecting LVSD were 83% (5/6), 94% (15/16), and 92% (24/26); and the negative predictive values were 99.8%, 99.0%, and 95.1%. Brain natriuretic peptide was not associated with LVSD in low-risk subjects (p = 0.087), but in IHD subjects (p = 0.015) and high-risk subjects (p = 0.023). Screening high-risk subjects by BNP before echocardiography could have reduced the cost per detected case of LVSD by 26% for the cost ratio of 1/20 (BNP/echocardiogram). CONCLUSIONS: Subjects at low and high risk of LVSD can be identified by simple clinical parameters, and BNP testing further reduces the number of echocardiograms needed and the costs of screening in subjects at risk <75 years of age in the general population. PMID- 12570954 TI - Limitation of cardiac output by total isovolumic time during pharmacologic stress in patients with dilated cardiomyopathy: activation-mediated effects of left bundle branch block and coronary artery disease. AB - OBJECTIVES: We sought to separate the effects of associated left bundle branch block (LBBB) and coronary artery disease (CAD) on peak cardiac output (CO) during dobutamine stress in patients with dilated cardiomyopathy (DCM). BACKGROUND: The mechanisms limiting CO during stress in patients with DCM are unclear. Both LBBB and CAD may do so by prolonging the total isovolumic time (t-IVT). METHODS: A total of 59 patients with DCM-34 with CAD (20 normal activation [NA], 14 LBBB) and 25 without CAD (15 NA, 10 LBBB)-were studied. The total IVT (s/min; calculated as: 60 - [total ejection time + total filling time] ) and CO were measured by Doppler echocardiography. RESULTS: At rest, t-IVT was 8 s/min longer with LBBB (p < 0.001), was unaffected by CAD, and did not correlate with rest CO. During stress, CO correlated with t-IVT (r = -0.73, p < 0.001) in all four patient groups. In the absence of CAD, t-IVT became shortened (NA by 7 +/- 3 s/min; LBBB by 9 +/- 4 s/min) and correlated with a fall in the QRS duration (NA: r = 0.87; LBBB: r = 0.91), and CO increased with stress (NA by 4.7 +/- 2.7 l/min; LBBB by 4.0 +/- 2.3 l/min; all p < 0.001). With CAD, t-IVT did not shorten normally with stress. Instead, t-IVT was 5.6 s/min longer and CO was 3.3 l/min lower than in those without CAD (both p < 0.001), and t-IVT did not correlate with the QRS duration. CONCLUSIONS: In patients with DCM, t-IVT during pharmacologic stress depends on changes in ventricular activation induced by LBBB or CAD and is, by itself, a major determinant of peak CO during stress. PMID- 12570955 TI - Prediction of mortality in patients with left ventricular hypertrophy by clinical, exercise stress, and echocardiographic data. AB - OBJECTIVES: This study evaluated the clinical, exercise stress test, and echocardiographic predictors of mortality and cardiac events in patients with left ventricular hypertrophy (LVH). BACKGROUND: Left ventricular hypertrophy is associated with an increased risk of cardiovascular morbidity and mortality. METHODS: Symptom-limited treadmill exercise echocardiography was performed for evaluation of coronary artery disease in 483 patients (age, 66 +/- 11 years; 281 men) with LVH. End points during follow-up were all-cause mortality and hard cardiac events (cardiac death and nonfatal myocardial infarction [MI]). RESULTS: Forty-six patients died and 14 had nonfatal MI. The cumulative mortality rate was higher in patients with abnormal exercise echocardiography (3% vs. 0.4% at one year, 11.7% vs. 3.7% at three years, and 18.3% vs. 9.5% at five years, p < 0.001). In a sequential multivariate analysis model of clinical, exercise test, and rest and exercise echocardiographic data, incremental predictors of mortality were workload (hazard ratio [HR], 0.5; 95% confidence interval [CI], 0.3 to 0.9), rate pressure product (HR, 0.7; 95% CI, 0.5 to 0.9), left ventricular (LV) mass index (HR, 1.4; 95% CI, 1.1 to 1.8), and failure to increase ejection fraction (EF) with exercise (HR, 2.1; 95% CI, 1.1 to 3.8). Predictors of cardiac events were history of coronary artery bypass grafting (HR, 2.6; 95% CI, 1.2 to 5.4), lower exercise rate-pressure product (HR, 0.6; 95% CI, 0.5 to 0.8), resting wall motion score index (HR, 1.4; 95% CI, 1.1 to 1.8), and failure to increase EF with exercise (HR, 3.3; 95% CI, 1.6 to 6.9). CONCLUSIONS: In patients with LVH, LV mass index and EF response to exercise are independent predictors of mortality, incremental to clinical and exercise test data and resting LV function. A normal exercise echocardiogram predicts a relatively low mortality rate during the following three years. PMID- 12570956 TI - Aortic valve sclerosis is associated with systemic endothelial dysfunction. AB - OBJECTIVES: We sought to examine the association between aortic valve sclerosis (AVS) and systemic endothelial manifestations of the atherosclerotic process. BACKGROUND: Clinical and experimental studies suggest that AVS is a manifestation of the atherosclerotic process. Systemic endothelial dysfunction is an early sign of the atherosclerotic process and can be assessed by ultrasonography of the brachial artery. METHODS: A total of 102 in-hospital patients (76 men; mean age 63.5 +/- 9.7 years) referred to the stress echocardiography laboratory underwent: 1) transthoracic echocardiography, with specific assessment of AVS (thickened valve leaflets with a transaortic flow velocity <2.5 m/s); 2) stress echocardiography; 3) coronary angiography, with evaluation of the Duke score (from 0 [normal] to 100 [most severe disease]); and 4) an endothelial function study, with assessment of endothelium-dependent, post-ischemic, flow-mediated dilation (FMD). RESULTS: Aortic valve sclerosis was present in 35 patients (group I) and absent in 67 (group II). Groups I and II were similar in terms of the frequency of stress-induced wall motion abnormalities (35.3% vs. 19.4%, p = NS) and the angiographic Duke score (33.8 +/- 28.6 vs. 35.2 +/- 29.1, p = NS). Patients with AVS showed a markedly lower FMD than those without AVS (2.2 +/- 3.5% vs. 5.3 +/- 5.3%, p < 0.01). On multivariate analysis, only FMD was highly predictive of AVS, with an odds ratio of 1.18 for each percent decrease in FMD (95% confidence interval 1.05 to 1.32; p = 0.01). CONCLUSIONS: Aortic valve stenosis is associated with systemic endothelial dysfunction. This observation may provide a mechanistic insight into the emerging association between AVS and cardiovascular events. PMID- 12570957 TI - Clinical efficacy of Doppler-echocardiographic indices of aortic valve stenosis: a comparative test-based analysis of outcome. AB - OBJECTIVES: This study was designed to assess which hemodynamic index best accounts for clinical severity of aortic stenosis (AS) and to analyze the value of low-dose dobutamine testing. BACKGROUND: Pressure gradient and valve area are suboptimal because they depend on flow rate, correlate poorly with symptoms, and provide limited prognostic information. Recently, new indices and low-dose inotropic stimulation have been introduced, but their clinical value remains uncertain. METHODS: A total of 307 consecutive patients with AS were included in an ambispective study design (71 +/- 12 years old; peak jet velocity: 3.7 +/- 1.1 m/s). Clinical and Doppler-echocardiographic data were obtained, as well as results of low-dose dobutamine infusion (47 patients). Using receiver-operator characteristic curve analysis, we evaluated jet velocity, pressure gradient, valve area, resistance, stroke-work loss (SWL), and dobutamine-induced increase in area for predicting 1) symptomatic status at entry, 2) early (25% best discriminated clinical end points. Other independent predictors of prognosis were symptomatic status and left ventricular ejection fraction. Dobutamine testing added no value to baseline models. CONCLUSIONS: Non-flow-corrected indices show the highest clinical efficacy in aortic stenosis. Among these, SWL best predicts symptomatic status and outcome and therefore should be incorporated to aid patient management in unclear situations. PMID- 12570958 TI - Aspergillus aortitis after cardiac surgery. AB - OBJECTIVES: The aim of this study was to describe the clinical characteristics of Aspergillus aortitis in a small series of consecutive patients. BACKGROUND: Aspergillus infection of the ascending aorta after cardiopulmonary bypass surgery has rarely been reported and has always resulted in death. METHODS: Aspergillus aortitis was confirmed by pathologic and microbiologic analysis in eight men (61 +/- 8 years) of 9,375 consecutive patients who underwent cardiac surgery between 1975 and 2000. RESULTS: Patients presented with Aspergillus aortitis after aortic valve replacement (n = 5), coronary revascularization (n = 2), or both (n = 1). Initial symptoms appeared between the immediate postoperative period and up to two years after surgery. All patients had prolonged fever. Ante-mortem diagnosis was established in only three patients for whom transthoracic echocardiography was suggestive of aortic pseudoaneurysm and was confirmed by thoracic computed tomography or aortography. All patients had negative peripheral blood cultures. Seven patients died at short-term follow-up, and the one surviving patient was promptly treated by surgery and antifungal drugs. Pathologic examination confirmed Aspergillus aortitis with multi-organ dissemination without heart involvement in all patients except for two, in whom aortic valve endocarditis was found. Fungal cultures confirmed the presence of Aspergillus fumigatus in all patients. CONCLUSIONS: Aspergillus aortitis is typically found after aortic valve or coronary surgery. It commonly leads to lethal multi-organ dissemination without involvement of the intracardiac structure. This entity should be considered in patients with persistent fever and negative blood cultures after open-heart surgery involving significant aortic wall damage, irrespective of the postoperative period. PMID- 12570959 TI - The elite reviewer. PMID- 12570960 TI - ACC/AHA 2002 guideline update for the management of patients with chronic stable angina--summary article: a report of the American College of Cardiology/American Heart Association Task Force on practice guidelines (Committee on the Management of Patients With Chronic Stable Angina). PMID- 12570961 TI - Immunosuppressive treatment in familial dilated cardiomyopathy with biopsy-proven intramyocardial inflammation? PMID- 12570963 TI - Left atrial appendectomy and maze. PMID- 12570965 TI - 'Let me explain': narrative emplotment and one patient's experience of oral cancer. AB - Recent research has investigated the way in which serious illness potentially poses a threat to peoples' sense of ontological security by throwing into doubt assumptions about time and the future. One of the main ways in which people adjust to such threats is through the use of narrative (either consciously or unconsciously) which helps to make sense of illness. Of particular relevance to people learning to live with a cancer diagnosis, is the concept of 'therapeutic emplotment' developed by Del Vecchio Good et al. (1994). This concept refers to the way in which oncologists are taught to structure temporal horizons for their patients in a particular way in order to instill and maintain hope in the context of arduous and toxic treatments. Using a case-study of one man's process of adapting to oral cancer (John Diamond's posthumously published serialised diary entries in The Times), this paper investigates the way in which such 'therapeutic emplotment' is implicitly incorporated by the patient, providing an underlying plot structure to his story. Following Diamond's diary entries over the 4 years duration of his illness, this paper analytically divides them into six main stages, documenting the underlying temporal structure and themes accompanying each stage of adaptation. The paper illustrates the way in which 'therapeutic emplotment' encourages the patient to focus on the immediate present and to place faith in the efficacy of specific treatments. However, it also explores how the attempt to live in the context of such a plot is fraught with anxiety for the patient, and how it co-exists with other largely 'unspoken narratives' of uncertainty, fear and skepticism in relation to the power of medicine. The main aim of the paper is to document, for the first time, the process of 'therapeutic emplotment' from the oral cancer patient's point of view. PMID- 12570966 TI - Moving from information transfer to information exchange in health and health care. AB - Communication in most health and health care settings assumes that information provision is both necessary and sufficient to improve individual behavior and, subsequently, health. This paper examines and challenges commonly accepted practices of information transmission in health settings, demonstrating how such practices are insufficient because they are rooted in a one-way model of information transfer. Three case studies show how this model is pervasive in different health and health care milieus: patient/provider encounters, health promotion programs, and national health policymaking. Drawing on critical theoretical perspectives, the work shows the limits of current information transfer approaches by critiquing the dominant assumptions that underpin current practice. At the same time, it provides empirical examples of the usefulness of critical approaches to identify relations of power in health communication. The paper concludes by suggesting that researchers and practitioners move beyond traditional practices of information transfer (based on a one-way monologue) and toward a more useful and appropriate notion of information exchange (based on two way dialogue). PMID- 12570967 TI - Norms, social networks, and HIV-related risk behaviors among urban disadvantaged drug users. AB - Altering norms may be an important approach to introducing and sustaining health protective behavior change. This study sought to examine the relationship between condom use, condom norms, and social network characteristics among a sample of economically impoverished individuals at risk for acquiring and transmitting HIV. Participants were 1051 individuals from a drug-using community in the USA. Eighty percent were current drug users; 17% were HIV seropositive. Reported condom use was strongly associated with peer norms about condom use (friends talking about condoms, encouraging condom use, and using condoms). Women were less likely than men to report that their friends used condoms. Injection drug use was negatively associated with peer norms about condom use, while church attendance and network characteristics were positively associated with condom-promoting norms. The size of the health advice and the financial support networks was most positively related to condom norms. Network methodology may aid in the identification of specific ties that promote condom use norms in a population. The findings of this study may have implications for norm change interventions among disadvantaged communities at high risk for HIV/AIDS. PMID- 12570968 TI - Tuberculosis mortality and recent childbirth: a retrospective case-control study of Gibraltarian women, 1874-1884. AB - Prior to the introduction of effective treatment for pulmonary tuberculosis, there was little consensus on the potential health risk of pregnancy among infected women. While, intuitively, pregnancy was viewed as a risk for tuberculosis disease and mortality, early studies could not establish such a link with any great certainty. Our case study combines the methods of family reconstitution and a case-control approach to explore the possibility that the physiological and social strains of recent childbirth and the early mothering of infants may have been risk factors in adult female tuberculosis mortality in late 19th-century Gibraltar. The study is based on 244 reproductive age women who died between 1874 and 1884; some 55% of these deaths were attributed to tuberculosis. The record linkage indicates that almost 12% of the women who died had given birth within the year preceding their death. Factoring in the effects of age at death, marital status, and religion, the logistic regression results indicate that recent childbirth did not increase the risk of tuberculosis mortality among these women. PMID- 12570969 TI - 'They look like my kind of people'--perceptions of smoking images in youth magazines. AB - Ten years ago research showed that pro-smoking images were prevalent in British youth magazines, whereas there was little coverage of smoking and health. Since then there has been a great expansion in the number of youth magazines, and particularly those aimed at young men. Titles such as FHM and Loaded, which did not exist at the time of the previous research, are now market leaders. New qualitative and quantitative research has therefore been conducted to examine the prevalence and impact of smoking images in this revitalised youth style magazines market. This paper presents the findings of the qualitative element of this research, which examined British first year students' perceptions of the pro smoking imagery found in the magazines and explored the relationship between this and their own smoking images and identities. The research found that: (a) this imagery was perceived to be, on the whole, attractive, sociable and reassuring; (b) that it supported young people's perceptions of smoking and reinforced their smoker identities; and (c) that it has the potential to be more powerful than advertising imagery. PMID- 12570970 TI - Re-reading the IPSS research record. AB - The International Pilot Study of Schizophrenia has had a major influence on mental health and research practice since its inception in the 1960s. It is most famous for demonstrating superior outcomes for people living with schizophrenia in developing countries over those living with schizophrenia in developed countries. Like other international research initiatives, it has been shaped by social, political and economic circumstances that contribute to positive and negative outcomes for participating nations. This paper evaluates the contribution of the pilot study by analyzing the discourse surrounding the dissemination of its results. The discussion of long-term outcomes across cultures is used to scrutinize the process of long-term research collaborations across nations. Discourse analysis of the research records indicates that, in various ways, the discourse was constructed to preserve an image of Western superiority and Third World inferiority. As the international research context shifts in psychiatry, it is hoped that inequitable practices that affect this and other studies will be challenged to benefit knowledge-building and mental health care across the world. PMID- 12570971 TI - Can the school make a difference? A multilevel analysis of adolescent risk and health behaviour. AB - The main aim of this article is to assess the relationship between the structural and (health) policy variables of the school and characteristics of the individual on the risk and health behaviour of adolescents. Individual and school level effects on seven health-related behaviours are simultaneously estimated, using multilevel modelling. The data are from the Flemish health behaviour in school aged children study in Belgium. Data are used from 29 Flemish schools in which students (N=3225), school administrators (N=29) and teachers (N=1132) were surveyed with anonymous written questions. The analysis confirms previous findings concerning individual level effects. Although differences between schools in risk and health behaviour were found to originate mainly from differences in pupil characteristics, substantial variation between schools remained with regard to regular smoking, drinking habits and tooth brushing after controlling for individual effects. A wide range of school structure and policy variables were taken into account, but only few of them were found to influence the health and risk behaviour of young people. Moreover, the study could not detect an effect of health promotion policy at school. The analysis therefore only partially confirms the hypothesis that the school has an impact on the health behaviour of young people. The findings demonstrate the need for a more thorough examination of the paths by which schools can influence the health behaviour of their pupils. PMID- 12570972 TI - Life in a secure unit: the rehabilitation of young people through the use of sport. AB - Sport is used widely in the rehabilitation of British young people who are resident in secure facilities following criminal sentencing, during remand or for welfare reasons. The research uses a quasi-longitudinal observational study in a secure unit, located in Southern England, and questions the suitability of sporting activities used as a primary vehicle for, and mechanism of, rehabilitation. Through the analysis of experiences at the unit, the paper supports the use of those sporting activities which de-emphasise regulations and winning, and argues for an emphasis on choice for participants, the tailoring of programmes to suit individual needs, and on positive feedback. Sport evidently has an important role in youth rehabilitation. However, it should be used selectively because, if unsuitable activities and programmes are used, they may have a detrimental effect on participants. The paper demonstrates the usefulness of qualitative research methods in rehabilitation studies. PMID- 12570973 TI - Impacts of an environmental disaster on psychosocial health and well-being in Karakalpakstan. AB - The people of Karakalpakstan, along with those of the entire Aral Sea region, are facing a multitude of health problems corresponding to the drying of the Aral Sea and accompanying ecological consequences. In case studies of other environmental disasters, research has shown that environmental exposures may impact not only the physiological but also the psychosocial health of individuals. This research aims to determine the contribution of the environmental disaster to the psychosocial health of people living in Karakalpakstan, a semi-autonomous Republic in Uzbekistan. An interview survey was carried out by Medecins Sans Frontieres, with the assistance of the McMaster Institute of Environment and Health, local Universities and local health care workers, on a random sample of 1118 individuals aged 18 years and older in three communities in Karakalpakstan in May/June 1999. The communities were chosen according to distance from the former seashore, urban/rural characteristics and ethnic composition. The survey included questions about perceived general health, the General Health Questionnaire, the somatic symptom checklist of the Symptom Check List-90, questions about perceptions of the environmental disaster, social support as well as socio-demographic and socio-economic characteristics. Findings show that 41% of all respondents reported environmental concern while 48% reported levels of somatic symptoms (SCL-90) associated with emotional distress, above the normalized cut-point. Significant differences in levels of emotional distress were reported between men and women as well as between ethnic groups. Environmental problems are commonly perceived to be the cause of somatic symptoms and are significantly related to self-rated health status. PMID- 12570974 TI - Perception of risk and subjective health among victims of the Chernobyl disaster. AB - Several studies have demonstrated that the nuclear power plant accident at Chernobyl in 1986 had a strong impact on the subjective health of the inhabitants in the surrounding regions and that the majority of these health complaints appear to be stress-related. An epidemiological survey among the adult population of the Gomel region in Belarus near Chernobyl showed higher rates of self reported health problems, psychological distress and medical service use in this region than in a comparable unexposed region. This paper presents an analysis of data on cognitive factors that were collected in this study. The findings support the hypothesis that cognitive variables such as risk perception and sense of control play an important role as mediating factors in the explanation of the observed health differences between the exposed and non-exposed regions. A tentative model is presented to further clarify the role of risk perception in the occurrence of non-specific health complaints after such ecological disasters. PMID- 12570975 TI - Causes and health consequences of environmental degradation and social injustice. AB - Worldwide the greatest effects on the health of individuals and populations results from environmental degradation and social injustice, operating in consort. This paper describes the national and global causes and health consequences of these phenomena. Causes include overpopulation, pollution, deforestation, global warming, unsustainable agricultural and fishing practices, overconsumption, maldistribution of wealth, the rise of the corporation, the Third World debt crisis, and militarization and wars. Consequences include increased poverty, overcrowding, famine, weather extremes, species loss, acute and chronic medical illnesses, war and human rights abuses, and an increasingly unstable global situation that portends Malthusian chaos and disaster. Because of their scientific training, and due to their privileged socioeconomic status, physicians are in a unique position to recognize these phenomena and to act at all levels, from interactions with their patients, to volunteerism, to service and intervention in areas of great need, to direct political activism and involvement. Specific suggestions for action are discussed. PMID- 12570976 TI - What are the ingredients for a successful evidence-based patient choice consultation?: A qualitative study. AB - The evidence-based patient choice (EBPC) approach is one of a number of newly emerging templates for medical encounters that advocate evidence-informed choice and shared decision-making. These models emphasise respect for patient preferences for involvement in health care decisions and advocate the sharing of good quality evidence-based information. In the medical consultation EBPC involves providing patients with evidence-based information in a way that facilitates their ability to make choices or decisions about their health care. Whereas the key principles of shared decision-making have been conceptualised, so far, no qualitative investigations have been undertaken to establish the key components of an EBPC consultation. Therefore, a series of semi-structured interviews were carried out with key informants to identify the elements and skills required for a successful EBPC consultation to occur. The interviews were conducted with purposively selected UK general practitioners (n=11), hospital doctors (n=10), practice nurses (n=5), academics (n=11) and lay people (n=8). Qualitative analysis of participants' responses was conducted using the constant comparative method. Six main themes emerged from the data, these were research evidence/medical information, the doctor-patient relationship, patient perspectives, decision-making processes, time issues and establishing the patient's problem. All respondents placed importance on doctors and patients being well informed and appraised of the latest available medical evidence. There was a general view that evidence-based information regarding diagnosis and treatment options should be shared with patients during a consultation. However, there were no suggestions as to how this might be achieved in practice. Participants' opinions relating to which model of decision-making should be adopted ranged from favouring an informed choice model, to the view that decision making should be shared equally. Similarly, there was no clear view on how much guidance a doctor should offer a patient during decision-making concerning the most appropriate treatment option for that patient. PMID- 12570977 TI - A certain art of uncertainty: case presentation and the development of professional identity. AB - Healthcare professionals use the genre of case presentation to communicate among themselves the salient patient information during treatment and management. In case presentation, many uncertainties surface, regarding, e.g., the reliability of patient reports, the sensitivity of laboratory tests, and the boundaries of scientific knowledge. The management and portrayal of uncertainty is a critical aspect of professional discourse. This paper documents the rhetorical features of certainty and uncertainty in novice case presentations, considering their pragmatic and problematic implications for students' professional socialization. This study was conducted during the third-year inpatient clerkship at a tertiary care, pediatric hospital in hospital in Canada. Data collection included: (1) non participant observations of 19 student case presentations involving 11 student and 10 faculty participants, and (2) individual interviews with 11 students and 10 faculty participants. A grounded theory approach informed data collection and analysis. Five thematic categories emerged, two of which this paper considers in detail: "Thinking as a Student" and "Thinking as a Doctor". Within these categories, the management and portrayal of uncertainty was a recurrent issue. Teachers modeled central features of a "professional rhetoric of uncertainty", managing uncertainty of six origins: limits of individual knowledge, limits of evidence, limitless possibility, limits of patient's/parent's account, limits of professional agreement, and limits of scientific knowledge. By contrast, students demonstrated a "novice rhetoric of uncertainty", represented by their focus on responding to personal knowledge deficits through the strategies of acknowledgement, argument, and deflection. Some students moved towards the professional rhetoric of uncertainty, suggesting not only advances in communication, but also shifts in attitude towards patients and colleagues, that were interpreted as indications that this rhetoric shapes professional identity and interactions. PMID- 12570978 TI - Sandwiching it in: spillover of work onto food choices and family roles in low- and moderate-income urban households. AB - Lower status jobs, high workloads and lack of control at work have been associated with less healthful diets, but the ways through which work is connected to food choices are not well understood. This analysis was an examination of workers' experience of the relationship of their jobs to their food choices. Fifty-one multi-ethnic, urban, low- and moderate-income adults living in Upstate New York in 1995 participated in a qualitative interview study of fruit and vegetable choices and discussed employment and food choices. The workers who participated in this study described a dynamic relationship between work and food choices that they experienced in the context of their other roles and values. These workers presented a relationship that was characterized by positive and negative spillover between their jobs and their ability to fulfill family roles and promote personal health, linked by a spectrum of food choice strategies. Participants' narratives fit into three different domains: characterizations of work and their resources for food choice, strategies used to manage food choices within the constraints of work, and affect related to the negative and positive spillover of these strategies on family roles and on personal food choices. Characterizations of work as demanding and limiting or demanding and manageable differentiated participants who experienced their food choice strategies as a source of guilt and dissatisfaction (negative spillover) from those who experienced food choices as a source of pride and satisfaction (positive spillover). Ideals and values related to food choice and health were balanced against other values for family closeness and nurturing and personal achievement. Some participants found work unproblematic. These findings direct attention to a broad conceptualization of the relationship of work to food choices in which the demands and resources of the work role are viewed as they spill over into the social and temporal context of other roles and values. PMID- 12570979 TI - Gender and coping: the parents of children with high functioning autism. AB - Gender is a concept that is frequently discussed in the literature on stress, coping and illness. Research has reported that women are more vulnerable than men are to stressful events and use different strategies to cope with them. Furthermore, it is often asserted that these gender-based differences in coping may partially explain the differential impact of stressful events on men and women. Unfortunately, much of this research has equated gender with sex and failed to contextualise the experience of illness and coping. This paper presents a qualitative analysis of the role of gender and coping among parents of children with high functioning autism or Asperger's syndrome in an Australian sample. It attempts to analyse the different meanings of the disability for mothers and fathers and describes the various strategies that parents use to cope with their child's disability. PMID- 12570980 TI - A model of mother-child coping and adjustment to HIV. AB - An increasing proportion of newly diagnosed AIDS cases is being reported among African American urban women. Recent research regarding the psychosocial and behavioral impact of a mother's HIV status on her uninfected children as well as a growing body of clinical evidence suggest that these children are extremely vulnerable and at risk for problems in psychosocial adjustment. The present paper reports the results of research designed to examine the pathways by which a mother's HIV-positive status affects the psychosocial adjustment of her uninfected school-age child. The principal predictor variables of the model are family sociodemographic characteristics, social support available to mother and child, HIV-related symptom distress in the mother, coping strategies of both mother and child, emotional distress of the mother, and quality of the parent child relationship. The dependent variable is the psychosocial adjustment of the child. Data were collected on 147 mother-child dyads using standardized questionnaires and personal interviews. Eighty-six percent of the mothers were African American and over 96% were on public assistance. Structural equation modeling was used to test the proposed model of mother-child coping and adjustment. After adding three paths, the model had a good fit to the data (comparative fit index=0.94; root mean square estimate of error=0.06). Five model constructs accounted for 36% of the variance in child adjustment. The constructs in order of importance were maternal HIV-associated stressors, maternal emotional distress, child social support, child coping, and quality of parent-child relationship. PMID- 12570981 TI - African-Caribbean interactions with mental health services in the UK: experiences and expectations of exclusion as (re)productive of health inequalities. AB - In the context of current concerns about health inequalities among minority ethnic groups in the UK, this paper addresses perceptions of mental health services among members of an African-Caribbean community in a South England town. Efforts to reduce health inequalities must take account of the views of local community members on the sources of those inequalities and on local health services. The statistical existence of inequalities in diagnosis and treatment of African-Caribbeans in the UK is well-established, supported by sociological explanations of these inequalities which centre on social exclusion in a variety of forms: institutional, cultural and socio-economic. However, detailed studies of the perspectives of local communities on mental health issues and services have received less attention. In this case study of community perceptions of mental health services, we find that social exclusion comprises an explanatory framework which is repeatedly invoked by community members in describing their interaction with mental health services. Interviewees assert that experience and expectation of racist mis-treatment by mental health services are key factors discouraging early accessing of mental health services, and thereby perpetuating mental health inequalities. We conclude that participation and partnership are vital means by which to generate both the objective and subjective inclusion that are requirements for an accessible and appropriate health service. PMID- 12570982 TI - Early activation of p160ROCK by pressure overload in rat heart. AB - We investigated the effects of acute pressure overload on activation of p160(ROCK) in rat myocardium. Constriction of transverse aorta, controlled to increase peak systolic pressure of ascending aorta by approximately 40 mmHg, induced a rapid association of RhoA with Dbl-3 and p160(ROCK). The binding of p160(ROCK) to RhoA was rapidly increased, peaking at 30 min (approximately 3.5 fold), but reduced to lower levels (approximately 1.9-fold) by 60 min of pressure overload. The activity of immunoprecipitated p160(ROCK) toward myosin light chain increased approximately 2.5-fold within 10 min but decreased to lower levels (approximately 1.6-fold) after 60 min of pressure overload. Confocal microscopic analysis indicated that pressure overload induced the formation of aggregates of p160(ROCK) and RhoA along the longitudinal axis of cardiac myocytes. Immunoelectron microscopic analysis showed that pressure overload induced the association of p160(ROCK) and RhoA to Z-line, T-tubule, and subsarcolemmal areas. The rapid activation of p160(ROCK) by pressure overload and its aggregation in subcellular structures involved in transmission of mechanical force suggest a role for this enzyme in the mechanobiochemical transduction in the myocardium. PMID- 12570983 TI - Na,K-ATPase inhibition alters tight junction structure and permeability in human retinal pigment epithelial cells. AB - Na,K-ATPase regulates a variety of transport functions in epithelial cells. In cultures of human retinal pigment epithelial (RPE) cells, inhibition of Na,K ATPase by ouabain and K(+) depletion decreased transepithelial electrical resistance (TER) and increased permeability of tight junctions to mannitol and inulin. Electrophysiological studies demonstrated that the decrease in TER was due to an increase in paracellular shunt conductance. At the light microscopy level, this increased permeability was not accompanied by changes in the localization of the tight junction proteins ZO-1, occludin, and claudin-3. At the ultrastructural level, increased tight junction permeability correlated with a decrease in tight junction membrane contact points. Decreased tight junction membrane contact points and increased tight junction permeability were reversible in K(+)-repletion experiments. Confocal microscopy revealed that in control cells, Na,K-ATPase was localized at both apical and basolateral plasma membranes. K(+) depletion resulted in a large reduction of apical Na,K-ATPase, and after K(+) repletion the apical Na,K-ATPase recovered to control levels. These results suggest a functional link exists between Na,K-ATPase and tight junction function in human RPE cells. PMID- 12570984 TI - Progressive decrease of intramyocellular accumulation of H+ and Pi in human skeletal muscle during repeated isotonic exercise. AB - The purpose of this study was to evaluate the hypotheses that accumulation of hydrogen ions and/or inorganic phosphate (Pi) in skeletal muscle increases with repeated bouts of isotonic exercise. (31)P-Magnetic resonance spectroscopy was used to examine the gastrocnemius muscle of seven highly aerobically trained females during four bouts of isotonic plantar flexion. The exercise bouts (EX1-4) of 3 min and 18 s were separated by 3 min and 54 s of complete rest. Muscle ATP did not change during the four bouts. Phosphocreatine (PCr) degradation during EX1 (13.3 +/- 2.4 mmol/kg wet weight) was higher (P < 0.01) compared with EX3-4 (9.7 +/- 1.6 and 9.6 +/- 1.8 mmol/kg wet weight, respectively). The intramyocellular pH at the end of EX1 (6.87 +/- 0.05) was significantly lower (P < 0.001) than those of EX2 (6.97 +/- 0.02), EX3 (7.02 +/- 0.01), and EX4 (7.02 +/ 0.02). Total Pi and diprotonated Pi were significantly higher (P < 0.001) at the end of EX1 (17.3 +/- 2.7 and 7.8 +/- 1.6 mmol/kg wet weight, respectively) compared with the values at the end of EX3 and EX4. The monoprotonated Pi at the end of EX1 (9.5 +/- 1.2 mmol/kg wet weight) was also significantly higher (P < 0.001) than that after EX4 (7.5 +/- 1.1 mmol/kg wet weight). Subjects' rating of perceived exertion increased (P < 0.001) toward exhaustion as the number of exercises progressed (7.1 +/- 0.4, EX1; 8.0 +/- 0.3, EX2; 8.5 +/- 0.3, EX3; and 9.0 +/- 0.4, EX4; scale from 0 to 10). The present results indicate that human muscle fatigue during repeated intense isotonic exercise is not due to progressive depletion of high energy phosphates nor to intracellular accumulation of hydrogen ions, total, mono-, or diprotonated Pi. PMID- 12570985 TI - Ethanol sensitivity of BK(Ca) channels from arterial smooth muscle does not require the presence of the beta 1-subunit. AB - Ethanol inhibition of large-conductance, Ca(2+)-activated K(+) (BK(Ca)) channels in aortic myocytes may contribute to the direct contraction of aortic smooth muscle produced by acute alcohol exposure. In this tissue, BK(Ca) channels consist of pore-forming (bslo) and modulatory (beta) subunits. Here, modulation of aortic myocyte BK(Ca) channels by acute alcohol was explored by expressing bslo subunits in Xenopus oocytes, in the absence and presence of beta(1) subunits, and studying channel responses to clinically relevant concentrations of ethanol in excised membrane patches. Overall, average values of bslo channel activity (NP(o), with N = no. of channels present in the patch; P(o) = probability of a single channel being open) in response to ethanol (3-200 mM) mildly decrease when compared with pre-ethanol, isosmotic controls. However, channel responses show qualitative heterogeneity at all ethanol concentrations. In the majority of patches (42/71 patches, i.e., 59%), a reversible reduction in NP(o) is observed. In this subset, the maximal effect is obtained with 100 mM ethanol, at which NP(o) reaches 46.2 +/- 9% of control. The presence of beta(1) subunits, which determines channel sensitivity to dihydrosoyaponin-I and 17beta estradiol, fails to modify ethanol action on bslo channels. Ethanol inhibition of bslo channels results from a marked increase in the mean closed time. Although the voltage dependence of gating remains unaffected, the apparent effectiveness of Ca(2+) to gate the channel is decreased by ethanol. These changes occur without modifications of channel conduction. In conclusion, a new molecular mechanism that may contribute to ethanol-induced aortic smooth muscle contraction has been identified and characterized: a functional interaction between ethanol and the bslo subunit and/or its lipid microenvironment, which leads to a decrease in BK(Ca) channel activity. PMID- 12570986 TI - Creatine uptake and creatine transporter expression among rat skeletal muscle fiber types. AB - Total creatine (Cr(total) = phosphocreatine + creatine) concentrations differ substantially among mammalian skeletal muscle. Because the primary means to add Cr(total) to muscle is uptake of creatine through the sodium-dependent creatine transporter (CrT), differences in creatine uptake and CrT expression could account for the variations in [Cr(total)] among muscle fiber types. To test this hypothesis, hindlimbs of adult rats were perfused with 0.05-1 mM [(14)C]creatine for up to 90 min. Creatine uptake rates at 1 mM creatine were greatest in the soleus (140 +/- 8.8 nmol x h(-1) x g(-1)), less in the red gastrocnemius (117 +/- 8.3), and least in the white gastrocnemius (97 +/- 10.7). These rates were unaltered by time, insulin concentration, or increased perfusate sodium concentration. Conversely, creatine uptake rates were correspondingly decreased among fiber types by lower creatine and sodium concentrations. The CrT protein content by Western blot analysis was similarly greatest in the soleus, less in the red gastrocnemius, and least in the white gastrocnemius, whereas CrT mRNA was not different. Creatine uptake rates differ among skeletal muscle fiber sections in a manner reasonably assigned to the 58-kDa band of the CrT. Furthermore, creatine uptake rates scale inversely with creatine content, with the lowest uptake rate in the fiber type with the highest Cr(total) and vice versa. This suggests that the creatine pool fractional turnover rate is not common across muscle phenotypes and, therefore, is differentially regulated. PMID- 12570987 TI - Nuclear CaMKII inhibits neuronal differentiation of PC12 cells without affecting MAPK or CREB activation. AB - Ca(2+)/calmodulin-regulated protein kinase II (CaMKII) mediates many cellular events. The four CaMKII isoforms have numerous splice variants, three of which contain nuclear localization signals. Little is known about the role of nuclear localized CaMKII in neuronal development. To study this process, PC12 cells were transfected to produce CaMKII targeted to either the cytoplasm or the nucleus and then treated with nerve growth factor (NGF). NGF triggers a signaling cascade (MAPK) that results in the differentiation of PC12 cells into a neuronal phenotype, marked by neurite outgrowth. The present study found that cells expressing nuclear targeted CaMKII failed to grow neurites, whereas cells expressing cytoplasmic CaMKII readily produced neurites. Inhibition of neuronal differentiation by nuclear CaMKII was independent of MAPK signaling, as sustained Erk phosphorylation was not affected. Phosphorylation of CREB was also unaffected. Thus nuclear CaMKII modifies neuronal differentiation by a mechanism independent of MAPK and CREB activation. PMID- 12570988 TI - Isolation and molecular characterization of pMG160, a mobilizable cryptic plasmid from Rhodobacter blasticus. AB - A 3.4-kb cryptic plasmid was obtained from a new isolate of Rhodobacter blasticus. This plasmid, designated pMG160, was mobilizable by the conjugative strain Escherichia coli S17.1 into Rhodobacter sphaeroides, Rhodobacter capsulatus, and Rhodopseudomonas palustris. It replicated in the latter strains but not in Rhodospirillum rubrum, Rhodocyclus gelatinosus, or Bradyrhizobium species. Plasmid pMG160 was stably maintained in R. sphaeroides for more than 100 generations in the absence of selection but showed segregational instability in R. palustris. Instability in R. palustris correlated with a decrease in plasmid copy number compared to the copy number in R. sphaeroides. The complete nucleotide sequence of plasmid pMG160 contained three open reading frames (ORFs). The deduced amino acid sequences encoded by ORF1 and ORF2 showed high degrees of homology to the MobS and MobL proteins that are involved in plasmid mobilization of certain plasmids. Based on homology with the Rep protein of several other plasmids, ORF3 encodes a putative rep gene initiator of plasmid replication. The functions of these sequences were demonstrated by deletion mapping, frameshift analysis, and analysis of point mutations. Two 6.1-kb pMG160-based E. coli-R. sphaeroides shuttle cloning vectors were constructed and designated pMG170 and pMG171. These two novel shuttle vectors were segregationally stable in R. sphaeroides growing under nonselective conditions. PMID- 12570989 TI - Cooperation between Lactococcus lactis and nonstarter lactobacilli in the formation of cheese aroma from amino acids. AB - In Gouda and Cheddar type cheeses the amino acid conversion to aroma compounds, which is a major process for aroma formation, is essentially due to lactic acid bacteria (LAB). In order to evaluate the respective role of starter and nonstarter LAB and their interactions in cheese flavor formation, we compared the catabolism of phenylalanine, leucine, and methionine by single strains and strain mixtures of Lactococcus lactis subsp. cremoris NCDO763 and three mesophilic lactobacilli. Amino acid catabolism was studied in vitro at pH 5.5, by using radiolabeled amino acids as tracers. In the presence of alpha-ketoglutarate, which is essential for amino acid transamination, the lactobacillus strains degraded less amino acids than L. lactis subsp. cremoris NCDO763, and produced mainly nonaromatic metabolites. L. lactis subsp. cremoris NCDO763 produced mainly the carboxylic acids, which are important compounds for cheese aroma. However, in the reaction mixture containing glutamate, only two lactobacillus strains degraded amino acids significantly. This was due to their glutamate dehydrogenase (GDH) activity, which produced alpha-ketoglutarate from glutamate. The combination of each of the GDH-positive lactobacilli with L. lactis subsp. cremoris NCDO763 had a beneficial effect on the aroma formation. Lactobacilli initiated the conversion of amino acids by transforming them mainly to keto and hydroxy acids, which subsequently were converted to carboxylic acids by the Lactococcus strain. Therefore, we think that such cooperation between starter L. lactis and GDH-positive lactobacilli can stimulate flavor development in cheese. PMID- 12570990 TI - Molecular analysis of a Saccharomyces cerevisiae mutant with improved ability to utilize xylose shows enhanced expression of proteins involved in transport, initial xylose metabolism, and the pentose phosphate pathway. AB - Differences between the recombinant xylose-utilizing Saccharomyces cerevisiae strain TMB 3399 and the mutant strain TMB 3400, derived from TMB 3399 and displaying improved ability to utilize xylose, were investigated by using genome wide expression analysis, physiological characterization, and biochemical assays. Samples for analysis were withdrawn from chemostat cultures. The characteristics of S. cerevisiae TMB 3399 and TMB 3400 grown on glucose and on a mixture of glucose and xylose, as well as of S. cerevisiae TMB 3400 grown on only xylose, were investigated. The strains were cultivated under chemostat conditions at a dilution rate of 0.1 h(-1), with feeds consisting of a defined mineral medium supplemented with 10 g of glucose liter(-1), 10 g of glucose plus 10 g of xylose liter(-1) or, for S. cerevisiae TMB 3400, 20 g of xylose liter(-1). S. cerevisiae TMB 3400 consumed 31% more xylose of a feed containing both glucose and xylose than S. cerevisiae TMB 3399. The biomass yields for S. cerevisiae TMB 3400 were 0.46 g of biomass g of consumed carbohydrate(-1) on glucose and 0.43 g of biomass g of consumed carbohydrate(-1) on xylose. A K(s) value of 33 mM for xylose was obtained for S. cerevisiae TMB 3400. In general, the percentage error was <20% between duplicate microarray experiments originating from independent fermentation experiments. Microarray analysis showed higher expression in S. cerevisiae TMB 3400 than in S. cerevisiae TMB 3399 for (i) HXT5, encoding a hexose transporter; (ii) XKS1, encoding xylulokinase, an enzyme involved in one of the initial steps of xylose utilization; and (iii) SOL3, GND1, TAL1, and TKL1, encoding enzymes in the pentose phosphate pathway. In addition, the transcriptional regulators encoded by YCR020C, YBR083W, and YPR199C were expressed differently in the two strains. Xylose utilization was, however, not affected in strains in which YCR020C was overexpressed or deleted. The higher expression of XKS1 in S. cerevisiae TMB 3400 than in TMB 3399 correlated with higher specific xylulokinase activity in the cell extracts. The specific activity of xylose reductase and xylitol dehydrogenase was also higher for S. cerevisiae TMB 3400 than for TMB 3399, both on glucose and on the mixture of glucose and xylose. PMID- 12570991 TI - Molecular characterization of astroviruses by reverse transcriptase PCR and sequence analysis: comparison of clinical and environmental isolates from South Africa. AB - A comparative analysis was performed with 25 isolates of astroviruses (AstVs) detected in sewage sources and 22 concurrently identified clinical AstV isolates from the Tshwane (Pretoria) Metropolitan Area in South Africa. The samples and specimens were screened for AstVs by using an enzyme immunoassay and/or a reverse transcriptase PCR (RT-PCR) for the highly conserved untranslated region (3' end) of the genome. The RT-PCR results were confirmed by oligonucleotide probe dot blot hybridization. Viable viruses were propagated in cell cultures for amplification when a minimal specimen was available or indeterminate sequences were obtained. AstV strains were characterized by RT-PCR and partial sequence analysis of the capsid region. The presence of multiple human AstV (HAstV) types in a single sewage sample complicated identification of individual strains, and additional type-specific RT-PCR and sequence analyses of the capsid region were required for characterization. Amplification and characterization of one genotype from a sample, therefore, did not preclude the possibility that a sample harbored additional different genotypes. Genotype and sequence information obtained from AstVs in wastewater samples were compared to information obtained from AstV strains from human stools. HAstV type 1 (HAstV-1), as well as HAstV-3, -5, -6, and -8, were identified among the clinical isolates, and HAstV-1, -2, -3, -4, -5, -7, and -8 were identified among the environmental samples. Phylogenetic analysis demonstrated that HAstV-1, -3, -5, and -8, which were present in human stool and sewage samples, clustered together, indicating that these viruses are closely related. The concurrent presence of identical HAstV strains in wastewater samples and in hospitalized patients suggests that AstVs present in the environment pose a potential risk to communities in which fecally contaminated water is used for recreational and domestic purposes. PMID- 12570992 TI - Identification of a high-affinity phosphate transporter gene in a prasinophyte alga, Tetraselmis chui, and its expression under nutrient limitation. AB - A high-affinity phosphate transporter gene, TcPHO, was isolated from a growth dependent subtracted cDNA library of the marine unicellular alga Tetraselmis chui. The full-length cDNA of TcPHO obtained by 5' and 3' rapid amplification of cDNA ends was 1,993 bp long and encoded an open reading frame consisting of 610 amino acids. The deduced amino acid sequence of TcPHO exhibited 51.6 and 49.8% similarity to the amino acid sequences of PHO89 from Saccharomyces cerevisiae and PHO4 from Neurospora crassa, respectively. In addition, hydrophobicity and secondary structure analyses revealed 12 conserved transmembrane domains that were the same as those found in PHO89 and PHO4. The expression of TcPHO mRNA was dependent on phosphate availability. With a low-phosphate treatment, the TcPHO mRNA concentration increased sharply to 2.72 fmol micro g of total RNA(-1) from day 1 to day 2 and remained at this high level from days 2 to 4. Furthermore, rescue treatment with either phosphate or p-nitrophenyl phosphate effectively inhibited TcPHO mRNA expression. In contrast, TcPHO mRNA expression stayed at a low level (range, 0.25 to 0.28 fmol micro g of total RNA(-1)) under low-nitrate conditions. The expression pattern suggests that TcPHO can be used as a molecular probe for monitoring phosphorus stress in T. chui. PMID- 12570993 TI - Anaerobic degradation of ethylbenzene by a new type of marine sulfate-reducing bacterium. AB - Anaerobic degradation of the aromatic hydrocarbon ethylbenzene was studied with sulfate as the electron acceptor. Enrichment cultures prepared with marine sediment samples from different locations showed ethylbenzene-dependent reduction of sulfate to sulfide and always contained a characteristic cell type that formed gas vesicles towards the end of growth. A pure culture of this cell type, strain EbS7, was isolated from sediment from Guaymas Basin (Gulf of California). Complete mineralization of ethylbenzene coupled to sulfate reduction was demonstrated in growth experiments with strain EbS7. Sequence analysis of the 16S rRNA gene revealed a close relationship between strain EbS7 and the previously described marine sulfate-reducing strains NaphS2 and mXyS1 (similarity values, 97.6 and 96.2%, respectively), which grow anaerobically with naphthalene and m xylene, respectively. However, strain EbS7 did not oxidize naphthalene, m-xylene, or toluene. Other compounds utilized by strain EbS7 were phenylacetate, 3 phenylpropionate, formate, n-hexanoate, lactate, and pyruvate. 1-Phenylethanol and acetophenone, the characteristic intermediates in anaerobic ethylbenzene degradation by denitrifying bacteria, neither served as growth substrates nor were detectable as metabolites by gas chromatography-mass spectrometry in ethylbenzene-grown cultures of strain EbS7. Rather, (1-phenylethyl)succinate and 4-phenylpentanoate were detected as specific metabolites in such cultures. Formation of these intermediates can be explained by a reaction sequence involving addition of the benzyl carbon atom of ethylbenzene to fumarate, carbon skeleton rearrangement of the succinate moiety (as a thioester), and loss of one carboxyl group. Such reactions are analogous to those suggested for anaerobic n alkane degradation and thus differ from the initial reactions in anaerobic ethylbenzene degradation by denitrifying bacteria which employ dehydrogenations. PMID- 12570994 TI - Monitoring gene expression in mixed microbial communities by using DNA microarrays. AB - A DNA microarray to monitor the expression of bacterial metabolic genes within mixed microbial communities was designed and tested. Total RNA was extracted from pure and mixed cultures containing the 2,4-dichlorophenoxyacetic acid (2,4-D) degrading bacterium Ralstonia eutropha JMP134, and the inducing agent 2,4-D. Induction of the 2,4-D catabolic genes present in this organism was readily detected 4, 7, and 24 h after the addition of 2,4-D. This strain was diluted into a constructed mixed microbial community derived from a laboratory scale sequencing batch reactor. Induction of two of five 2,4-D catabolic genes (tfdA and tfdC) from populations of JMP134 as low as 10(5) cells/ml was clearly detected against a background of 10(8) cells/ml. Induction of two others (tfdB and tfdE) was detected from populations of 10(6) cells/ml in the same background; however, the last gene, tfdF, showed no significant induction due to high variability. In another experiment, the induction of resin acid degradative genes was statistically detectable in sludge-fed pulp mill effluent exposed to dehydroabietic acid in batch experiments. We conclude that microarrays will be useful tools for the detection of bacterial gene expression in wastewaters and other complex systems. PMID- 12570995 TI - Hydrogen-dependent oxygen reduction by homoacetogenic bacteria isolated from termite guts. AB - Although homoacetogenic bacteria are generally considered to be obligate anaerobes, they colonize the intestinal tracts of termites and other environments that are not entirely anoxic in space or time. In this study, we investigated how homoacetogenic bacteria isolated from the hindguts of various termites respond to the presence of molecular oxygen. All strains investigated formed growth bands in oxygen gradient agar tubes under a headspace of H(2)-CO(2). The position of the bands coincided with the oxic-anoxic interface and depended on the O(2) partial pressure in the headspace; the position of the bands relative to the meniscus remained stable for more than 1 month. Experiments with dense cell suspensions, performed with Clark-type O(2) and H(2) electrodes, revealed a large capacity for H(2)-dependent oxygen reduction in Sporomusa termitida and Sporomusa sp. strain TmAO3 (149 and 826 nmol min(-1) mg of protein(-1), respectively). Both strains also reduced O(2) with endogenous reductants, albeit at lower rates. Only in Acetonema longum did the basal rates exceed the H(2)-dependent rates considerably (181 versus 28 nmol min(-1) mg of protein)(-1)). Addition of organic substrates did not stimulate O(2) consumption in any of the strains. Nevertheless, reductive acetogenesis by cell suspensions of strain TmAO3 was inhibited even at the lowest O(2) fluxes, and growth in nonreduced medium occurred only after the bacteria had rendered the medium anoxic. Similar results were obtained with Acetobacterium woodii, suggesting that the results are not unique to the strains isolated from termites. We concluded that because of their tolerance to temporary exposure to O(2) at low partial pressures (up to 1.5 kPa in the case of strain TmAO3) and because of their large capacity for O(2) reduction, homoacetogens can reestablish conditions favorable for growth by actively removing oxygen from their environment. PMID- 12570996 TI - The abundance of microcystin-producing genotypes correlates positively with colony size in Microcystis sp. and determines its microcystin net production in Lake Wannsee. AB - The working hypotheses tested on a natural population of Microcystis sp. in Lake Wannsee (Berlin, Germany) were that (i) the varying abundance of microcystin producing genotypes versus non-microcystin-producing genotypes is a key factor for microcystin net production and (ii) the occurrence of a gene for microcystin net production is related to colony morphology, particularly colony size. To test these hypotheses, samples were fractionated by colony size with a sieving procedure during the summer of 2000. Each colony size class was analyzed for cell numbers, the proportion of microcystin-producing genotypes, and microcystin concentrations. The smallest size class of Microcystis colonies (<50 microm) showed the lowest proportion of microcystin-producing genotypes, the highest proportion of non-microcystin-producing cells, and the lowest microcystin cell quotas (sum of microcystins RR, YR, LR, and WR). In contrast, the larger size classes of Microcystis colonies (>100 microm) showed the highest proportion of microcystin-producing genotypes, the lowest proportion of non-microcystin producing cells, and the highest microcystin cell quotas. The microcystin net production rate was nearly one to one positively related to the population growth rate for the larger colony size classes (>100 microm); however, no relationship could be found for the smaller size classes. It was concluded that the variations found in microcystin net production between colony size classes are chiefly due to differences in genotype composition and that the microcystin net production in the lake is mainly influenced by the abundance of the larger (>100- microm) microcystin-producing colonies. PMID- 12570997 TI - Characterization of the initial reactions during the cometabolic oxidation of methyl tert-butyl ether by propane-grown Mycobacterium vaccae JOB5. AB - The initial reactions in the cometabolic oxidation of the gasoline oxygenate, methyl tert-butyl ether (MTBE), by Mycobacterium vaccae JOB5 have been characterized. Two products, tert-butyl formate (TBF) and tert-butyl alcohol (TBA), rapidly accumulated extracellularly when propane-grown cells were incubated with MTBE. Lower rates of TBF and TBA production from MTBE were also observed with cells grown on 1- or 2-propanol, while neither product was generated from MTBE by cells grown on casein-yeast extract-dextrose broth. Kinetic studies with propane-grown cells demonstrated that TBF is the dominant (> or = 80%) initial product of MTBE oxidation and that TBA accumulates from further biotic and abiotic hydrolysis of TBF. Our results suggest that the biotic hydrolysis of TBF is catalyzed by a heat-stable esterase with activity toward several other tert-butyl esters. Propane-grown cells also oxidized TBA, but no further oxidation products were detected. Like the oxidation of MTBE, TBA oxidation was fully inhibited by acetylene, an inactivator of short-chain alkane monooxygenase in M. vaccae JOB5. Oxidation of both MTBE and TBA was also inhibited by propane (K(i) = 3.3 to 4.4 microM). Values for K(s) of 1.36 and 1.18 mM and for V(max) of 24.4 and 10.4 nmol min(-1) mg of protein(-1) were derived for MTBE and TBA, respectively. We conclude that the initial steps in the pathway of MTBE oxidation by M. vaccae JOB5 involve two reactions catalyzed by the same monooxygenase (MTBE and TBA oxidation) that are temporally separated by an esterase-catalyzed hydrolysis of TBF to TBA. These results that suggest the initial reactions in MTBE oxidation by M. vaccae JOB5 are the same as those that we have previously characterized in gaseous alkane-utilizing fungi. PMID- 12570998 TI - A sensitive and inexpensive yeast bioassay for the mycotoxin zearalenone and other compounds with estrogenic activity. AB - Zearalenone (ZON) is a nonsteroidal estrogenic mycotoxin produced by plant pathogenic species of Fusarium. As a consequence of infection with Fusarium culmorum and Fusarium graminearum, ZON can be found in cereals and derived food products. Since ZON is suspected to be a cause of human disease, including premature puberty syndrome, as well as hyperestrogenism in farm animals, several countries have established monitoring programs and guidelines for ZON levels in grain intended for human consumption and animal feed. We developed a low-cost method for monitoring ZON contamination in grain based on a sensitive yeast bioassay. The indicator Saccharomyces cerevisiae strain YZRM7 is unable to grow unless an engineered pyrimidine biosynthetic gene is activated by the expressed human estrogen receptor in the presence of exogenous estrogenic substances. Deletion of the genes encoding ATP-binding cassette (ABC) transporters Pdr5p and Snq2p increases net ZON uptake synergistically. Less than 1 microg of ZON per liter of medium is sufficient to allow growth of the indicator strain. To prevent interference with pyrimidines potentially present in biological samples, we also disrupted the genes FUR1 and URK1, blocking the pyrimidine salvage pathway. The bioassay strain YZRM7 allows qualitative detection and quantification of total estrogenic activity in cereal extracts without requiring further cleanup steps. Its high sensitivity makes this assay suitable for low-cost monitoring of contamination of maize and small grain cereals with estrogenic Fusarium mycotxins. PMID- 12570999 TI - Integrative transformation system for the metabolic engineering of the sphingoid base-producing yeast Pichia ciferrii. AB - We have developed an integrative transformation system for metabolic engineering of the tetraacetyl phytosphingosine (TAPS)-secreting yeast Pichia ciferrii. The system uses (i) a mutagenized ribosomal protein L41 gene of P. ciferrii as a dominant selection marker that confer resistance to the antibiotic cycloheximide and (ii) a ribosomal DNA (rDNA) fragment of P. ciferrii as a target for multicopy gene integration into the chromosome. A locus within the nontranscribed region located between 5S and 26S rDNAs was selected as the integration site. A maximum frequency of integrative transformation of approximately 1,350 transformants/ microg of DNA was observed. To improve the de novo synthesis of sphingolipid, the LCB2 gene, encoding a subunit of serine palmitoyltransferase, which catalyzes the first committed step of sphingolipid synthesis, was cloned from P. ciferrii and overexpressed under the control of the P. ciferrii glyceraldehyde-3-phosphate dehydrogenase promoter. After transformation of an LCB2 gene expression cassette, several transformants that contained approximately five to seven copies of transforming DNA in the chromosome and exhibited about 50-fold increase in LCB2 mRNA relative to the wild type were identified. These transformants were observed to produce approximately two times more TAPS than the wild type. PMID- 12571000 TI - Contribution of pilA to competitive colonization of the squid Euprymna scolopes by Vibrio fischeri. AB - Vibrio fischeri colonizes the squid Euprymna scolopes in a mutualistic symbiosis. Hatchling squid lack these bacterial symbionts, and V. fischeri strains must compete to occupy this privileged niche. We cloned a V. fischeri gene, designated pilA, that contributes to colonization competitiveness and encodes a protein similar to type IV-A pilins. Unlike its closest known relatives, Vibrio cholerae mshA and vcfA, pilA is monocistronic and not clustered with genes associated with pilin export or assembly. Using wild-type strain ES114 as the parent, we generated an in-frame pilA deletion mutant, as well as pilA mutants marked with a kanamycin resistance gene. In mixed inocula, marked mutants were repeatedly outcompeted by ES114 (P < 0.05) but not by an unmarked pilA mutant, for squid colonization. In contrast, the ratio of mutant to ES114 CFUs did not change during 70 generations of coculturing. The competitive defect of pilA mutants ranged from 1.7- to 10-fold and was more pronounced when inocula were within the range estimated for V. fischeri populations in Hawaiian seawater (200 to 2,000 cells/ml) than when higher densities were used. ES114 also outcompeted a pilA mutant by an average of twofold at lower inoculum densities, when only a fraction of the squid became infected, most by only one strain. V. fischeri strain ET101, which was isolated from Euprymna tasmanica and is outcompeted by ES114, lacks pilA; however, 11 other diverse V. fischeri isolates apparently possess pilA. The competitive defect of pilA mutants suggests that cell surface molecules may play important roles in the initiation of beneficial symbioses in which animals must acquire symbionts from a mixed community of environmental bacteria. PMID- 12571001 TI - In-field spatial variability in the degradation of the phenyl-urea herbicide isoproturon is the result of interactions between degradative Sphingomonas spp. and soil pH. AB - Substantial spatial variability in the degradation rate of the phenyl-urea herbicide isoproturon (IPU) [3-(4-isopropylphenyl)-1,1-dimethylurea] has been shown to occur within agricultural fields, with implications for the longevity of the compound in the soil, and its movement to ground- and surface water. The microbial mechanisms underlying such spatial variability in degradation rate were investigated at Deep Slade field in Warwickshire, United Kingdom. Most-probable number analysis showed that rapid degradation of IPU was associated with proliferation of IPU-degrading organisms. Slow degradation of IPU was linked to either a delay in the proliferation of IPU-degrading organisms or apparent cometabolic degradation. Using enrichment techniques, an IPU-degrading bacterial culture (designated strain F35) was isolated from fast-degrading soil, and partial 16S rRNA sequencing placed it within the Sphingomonas group. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified bacterial community 16S rRNA revealed two bands that increased in intensity in soil during growth-linked metabolism of IPU, and sequencing of the excised bands showed high sequence homology to the Sphingomonas group. However, while F35 was not closely related to either DGGE band, one of the DGGE bands showed 100% partial 16S rRNA sequence homology to an IPU-degrading Sphingomonas sp. (strain SRS2) isolated from Deep Slade field in an earlier study. Experiments with strains SRS2 and F35 in soil and liquid culture showed that the isolates had a narrow pH optimum (7 to 7.5) for metabolism of IPU. The pH requirements of IPU-degrading strains of Sphingomonas spp. could largely account for the spatial variation of IPU degradation rates across the field. PMID- 12571002 TI - Microbial community composition affects soil fungistasis. AB - Most soils inhibit fungal germination and growth to a certain extent, a phenomenon known as soil fungistasis. Previous observations have implicated microorganisms as the causal agents of fungistasis, with their action mediated either by available carbon limitation (nutrient deprivation hypothesis) or production of antifungal compounds (antibiosis hypothesis). To obtain evidence for either of these hypotheses, we measured soil respiration and microbial numbers (as indicators of nutrient stress) and bacterial community composition (as an indicator of potential differences in the composition of antifungal components) during the development of fungistasis. This was done for two fungistatic dune soils in which fungistasis was initially fully or partly relieved by partial sterilization treatment or nutrient addition. Fungistasis development was measured as restriction of the ability of the fungi Chaetomium globosum, Fusarium culmorum, Fusarium oxysporum, and Trichoderma harzianum to colonize soils. Fungistasis did not always reappear after soil treatments despite intense competition for carbon, suggesting that microbial community composition is important in the development of fungistasis. Both microbial community analysis and in vitro antagonism tests indicated that the presence of pseudomonads might be essential for the development of fungistasis. Overall, the results lend support to the antibiosis hypothesis. PMID- 12571003 TI - Computer simulation of Clostridium botulinum strain 56A behavior at low spore concentrations. AB - It is generally assumed that spore behavior is independent of spore concentration, but recently published mathematical models indicate that this is not the case. A Monte Carlo simulation was employed in this study to further examine the independence assumption by evaluating the inherent variance in spore germination data. All simulations were carried out with @Risk software. A total of 500 to 4,000 iterations were needed for each simulation to reach convergence. Lag time and doubling time from a higher inoculum concentration were used to simulate the time to detection (TTD) at a lower inoculum concentration under otherwise identical environmental conditions. The point summaries of the simulated and observed TTDs were recorded for the 26 simulations, with kinetic data at the target inoculum concentration. The ratios of the median (R(m) = median(obs)/median(sim)) and 90% range (R(r) = 90% range(obs)/90% range(sim)) were calculated. Most R(m) and R(r) values were greater than one, indicating that the simulated TTDs were smaller and more homogeneous than the observed ones. R(r) values departed farther from one than R(m) values. Ratios obtained when simulating 1 spore with 10,000 spores deviated the farthest from one. Neither ratio was significantly different from the other when simulating 1 spore with 100 spores or simulating 100 spores with 10,000 spores. When kinetic data were not available, the percent positive observed at the 95th percentile of the simulated TTDs was obtained. These simulation results confirmed that the assumption of independence between spores is not valid. PMID- 12571004 TI - Bacterial population changes in a membrane bioreactor for graywater treatment monitored by denaturing gradient gel electrophoretic analysis of 16S rRNA gene fragments. AB - The bacterial population of a graywater treatment system was monitored over the course of 100 days, along with several wastewater biochemical parameters. The graywater treatment system employed an 1,800-liter membrane bioreactor (MBR) to process the waste, with essentially 100% recycling of the biomass. Graywater feed consisting of 10% galley water and 90% laundry water, selected to approximate the graywater composition on board U.S. Navy ships, was collected offsite. Five-day biological oxygen demand (BOD(5)), oils and greases (O/G), nitrogen, and phosphorus were monitored in the feed and were found to vary greatly day to day. Changes in the bacterial population were monitored by PCR amplification of region 332 to 518 (Escherichia coli numbering) of the 16S rRNA gene and denaturing gradient gel electrophoresis (DGGE) analysis of the resultant PCR products. DGGE analysis indicated a diverse and unstable bacterial population throughout the 100 day period, with spikes in feed strength causing significant changes in community structure. Long-term similarity between the communities was 0 to 25%, depending on the method of analysis. In spite of the unstable bacterial population, the MBR system was able to meet effluent quality parameters approximately 90% of the time. PMID- 12571005 TI - Production of cyclic lipopeptides by Pseudomonas fluorescens strains in bulk soil and in the sugar beet rhizosphere. AB - The production of cyclic lipopeptides (CLPs) with antifungal and biosurfactant properties by Pseudomonas fluorescens strains was investigated in bulk soil and in the sugar beet rhizosphere. Purified CLPs (viscosinamide, tensin, and amphisin) were first shown to remain highly stable and extractable (90%) when applied (ca. 5 microg g(-1)) to sterile soil, whereas all three compounds were degraded over 1 to 3 weeks in nonsterile soil. When a whole-cell inoculum of P. fluorescens strain DR54 containing a cell-bound pool of viscosinamide was added to the nonsterile soil, declining CLP concentrations were observed over a week. By comparison, addition of the strains 96.578 and DSS73 without cell-bound CLP pools did not result in detectable tensin or amphisin in the soil. In contrast, when sugar beet seeds were coated with the CLP-producing strains and subsequently germinated in nonsterile soil, strain DR54 maintained a high and constant viscosinamide level in the young rhizosphere for approximately 2 days while strains 96.578 and DSS73 exhibited significant production (net accumulation) of tensin or amphisin, reaching a maximum level after 2 days. All three CLPs remained detectable for several days in the rhizosphere. Subsequent tests of five other CLP-producing P. fluorescens strains also demonstrated significant production in the young rhizosphere. The results thus provide evidence that production of different CLPs is a common trait among many P. fluorescens strains in the soil environment, and further, that the production is taking place only in specific habitats like the rhizosphere of germinating sugar beet seeds rather than in the bulk soil. PMID- 12571006 TI - Production by Clostridium acetobutylicum ATCC 824 of CelG, a cellulosomal glycoside hydrolase belonging to family 9. AB - The genome sequence of Clostridium acetobutylicum ATCC 824, a noncellulolytic solvent-producing strain, predicts the production of various proteins with domains typical for cellulosomal subunits. Most of the genes coding for these proteins are grouped in a cluster similar to that found in cellulolytic clostridial species, such as Clostridium cellulovorans. CAC0916, one of the open reading frames present in the putative cellulosome gene cluster, codes for CelG, a putative endoglucanase belonging to family 9, and it was cloned and overexpressed in Escherichia coli. The overproduced CelG protein was purified by making use of its high affinity for cellulose and was characterized. The biochemical properties of the purified CelG were comparable to those of other known enzymes belonging to the same family. Expression of CelG by C. acetobutylicum grown on different substrates was studied by Western blotting by using antibodies raised against the purified E. coli-produced protein. Whereas the antibodies cross-reacted with CelG-like proteins secreted by cellobiose- or cellulose-grown C. cellulovorans cultures, CelG was not detectable in extracellular medium from C. acetobutylicum grown on cellobiose or glucose. However, notably, when lichenan-grown cultures were used, several bands corresponding to CelG or CelG-like proteins were present, and there was significantly increased extracellular endoglucanase activity. PMID- 12571007 TI - Tin-carbon cleavage of organotin compounds by pyoverdine from Pseudomonas chlororaphis. AB - The triphenyltin (TPT)-degrading bacterium Pseudomonas chlororaphis CNR15 produces extracellular yellow substances to degrade TPT. Three substances (F-I, F IIa, and F-IIb) were purified, and their structural and catalytic properties were characterized. The primary structure of F-I was established using two-dimensional nuclear magnetic resonance techniques; the structure was identical to that of suc pyoverdine from P. chlororaphis ATCC 9446, which is a peptide siderophore produced by fluorescent pseudomonads. Spectral and isoelectric-focusing analyses revealed that F-IIa and F-IIb were also pyoverdines, differing only in the acyl substituent attached to the chromophore part of F-I. Furthermore, we found that the fluorescent pseudomonads producing pyoverdines structurally different from F I showed TPT degradation activity in the solid extracts of their culture supernatants. F-I and F-IIa degraded TPT to monophenyltin via diphenyltin (DPT) and degraded DPT and dibutyltin to monophenyltin and monobutyltin, respectively. The total amount of organotin metabolites produced by TPT degradation was nearly equivalent to that of the F-I added to the reaction mixture, whereas DPT degradation was not influenced by monophenyltin production. The TPT degradation activity of F-I was remarkably inhibited by the addition of metal ions chelated with pyoverdine. On the other hand, the activity of DPT was increased 13- and 8 fold by the addition of Cu(2+) and Sn(4+), respectively. These results suggest that metal-chelating ligands common to pyoverdines may play important roles in the Sn-C cleavage of organotin compounds in both the metal-free and metal complexed states. PMID- 12571008 TI - Rhizobium etli and Rhizobium gallicum nodulate common bean (Phaseolus vulgaris) in a traditionally managed milpa plot in Mexico: population genetics and biogeographic implications. AB - The stability of the genetic structure of rhizobial populations nodulating Phaseolus vulgaris cultivated in a traditionally managed milpa plot in Mexico was studied over three consecutive years. The set of molecular markers analyzed (including partial rrs, glnII, nifH, and nodB sequences), along with host range experiments, placed the isolates examined in Rhizobium etli bv. phaseoli and Rhizobium gallicum bv. gallicum. Cluster analysis of multilocus enzyme electrophoresis and plasmid profile data separated the two species and identified numerically dominant clones within each of them. Population genetic analyses showed that there was high genetic differentiation between the two species and that there was low intrapopulation differentiation of the species over the 3 years. The results of linkage disequilibrium analyses are consistent with an epidemic genetic structure for both species, with frequent genetic exchange taking place within conspecific populations but not between the R. etli and R. gallicum populations. A subsample of isolates was selected and used for 16S ribosomal DNA PCR-restriction fragment length polymorphism analysis, nifH copy number determination, and host range experiments. Plasmid profiles and nifH hybridization patterns also revealed the occurrence of lateral plasmid transfer among distinct multilocus genotypes within species but not between species. Both species were recovered from nodules of the same plants, indicating that mechanisms other than host, spatial, or temporal isolation may account for the genetic barrier between the species. The biogeographic implications of finding an R. gallicum bv. gallicum population nodulating common bean in America are discussed. PMID- 12571009 TI - Characterization of chitinase genes from an alkaliphilic actinomycete, Nocardiopsis prasina OPC-131. AB - An alkaliphilic actinomycete, Nocardiopsis prasina OPC-131, secretes chitinases, ChiA, ChiB, and ChiB Delta, in the presence of chitin. The genes encoding ChiA and ChiB were cloned and sequenced. The open reading frame (ORF) of chiA encoded a protein of 336 amino acids with a calculated molecular mass of 35,257 Da. ChiA consisted of only a catalytic domain and showed a significant homology with family 18 chitinases. The chiB ORF encoded a protein of 296 amino acids with a calculated molecular mass of 31,500 Da. ChiB is a modular enzyme consisting of a chitin-binding domain type 3 (ChtBD type 3) and a catalytic domain. The catalytic domain of ChiB showed significant similarity to Streptomyces family 19 chitinases. ChiB Delta was the truncated form of ChiB lacking ChtBD type 3. Expression plasmids coding for ChiA, ChiB, and ChiB Delta were constructed to investigate the biochemical properties of these recombinant proteins. These enzymes showed pHs and temperature optima similar to those of native enzymes. ChiB showed more efficient hydrolysis of chitin and stronger antifungal activity than ChiB Delta, indicating that the ChtBD type 3 of ChiB plays an important role in the efficient hydrolysis of chitin and in antifungal activity. Furthermore, the finding of family 19 chitinase in N. prasina OPC-131 suggests that family 19 chitinases are distributed widely in actinomycetes other than the genus Streptomyces. PMID- 12571010 TI - Evaluation of broiler litter with reference to the microbial composition as assessed by using 16S rRNA and functional gene markers. AB - Very little is known about the microbial composition of animal bedding wastes, including poultry litter, and what is known has been deduced from standard culture methods, by which some fastidious organisms that exist in the environment may not be detected. We evaluated the bacterial composition of poultry litter by using a combination of culture and molecular detection. Total aerobic bacteria in poultry litter were detected by culture at 10(9) CFU/g of material. Enteric bacteria such as Enterococcus spp. and coliforms composed 0.1 and 0.01%, respectively, of the total aerobic cultivatable bacteria in poultry litter; no Salmonella strains were detected by culture. In order to characterize the most abundant bacterial groups, we sequenced 16S ribosomal DNA (rDNA) genes amplified by PCR with microbial community DNA isolated from poultry litter as the template. From the 16S rDNA library, 31 genera were identified. Twelve families or groups were identified with lactobacilli and Salinococcus spp. forming the most abundant groups. In fact, 82% of the total sequences were identified as gram-positive bacteria with 62% of total belonging to low G+C gram-positive groups. In addition to detection of 16S rDNA sequences associated with the expected fecal bacteria present in manure, we detected many bacterial sequences for organisms, such as Globicatella sulfidofaciens, Corynebacterium ammoniagenes, Corynebacterium urealyticum, Clostridium aminovalericum, Arthrobacter sp., and Denitrobacter permanens, that may be involved in the degradation of wood and cycling of nitrogen and sulfur. Several sequences were identified in the library for bacteria associated with disease in humans and poultry such as clostridia, staphylococci, and Bordetella spp. However, specific PCR targeting other human and veterinary pathogens did not detect the presence of Salmonella, pathogenic Escherichia coli, Campylobacter spp., Yersinia spp., Listeria spp., or toxigenic staphylococci. PCR and DNA hybridization revealed the presence of class 1 integrons with gene cassettes that specify resistance to aminoglycosides and chloramphenicol. Only from understanding the microbial community of animal wastes such as poultry litter can we manage animal disease and limit the impact of animal waste on the environment and human and animal health. PMID- 12571011 TI - Arthrobacter strain VAI-A utilizes acyl-homoserine lactone inactivation products and stimulates quorum signal biodegradation by Variovorax paradoxus. AB - Many Proteobacteria produce acyl-homoserine lactones (acyl-HSLs) and employ them as dedicated cell-to-cell signals in a process known as quorum sensing. Previously, Variovorax paradoxus VAI-C was shown to utilize diverse acyl-HSLs as sole sources of energy and nitrogen. We describe here the properties of a second isolate, Arthrobacter strain VAI-A, obtained from the same enrichment culture that yielded V. paradoxus VAI-C. Although strain VAI-A grew rapidly and exponentially on a number of substrates, it grew only slowly and aberrantly (i.e., linearly) in media amended with oxohexanoyl-HSL as the sole energy source. Increasing the culture pH markedly improved the growth rate in media containing this substrate but did not abolish the aberrant kinetics. The observed growth was remarkably similar to the known kinetics of the pH-influenced half-life of acyl HSLs, which decay chemically to yield the corresponding acyl-homoserines. Strain VAI-A grew rapidly and exponentially when provided with an acyl-homoserine as the sole energy or nitrogen source. The isolate was also able to utilize HSL as a sole source of nitrogen but not as energy for growth. V. paradoxus, known to release HSL as a product of quorum signal degradation, was examined for the ability to support the growth of Arthrobacter strain VAI-A in defined cocultures. It did. Moreover, the acyl-HSL-dependent growth rate and yield of the coculture were dramatically superior to those of the monocultures. This suggested that the original coenrichment of these two organisms from the same soil sample was not coincidental and that consortia may play a role in quorum signal turnover and mineralization. The fact that Arthrobacter strain VAI-A utilizes the two known nitrogenous degradation products of acyl-HSLs, acyl-homoserine and HSL, begins to explain why none of the three compounds are known to accumulate in the environment. PMID- 12571012 TI - Heat and osmotic stress responses of probiotic Lactobacillus rhamnosus HN001 (DR20) in relation to viability after drying. AB - The viability of lactic acid bacteria in frozen, freeze-dried, and air-dried forms is of significant commercial interest to both the dairy and food industries. In this study we observed that when prestressed with either heat (50 degrees C) or salt (0.6 M NaCl), Lactobacillus rhamnosus HN001 (also known as DR20) showed significant (P < 0.05) improvement in viability compared with the nonstressed control culture after storage at 30 degrees C in the dried form. To investigate the mechanisms underlying this stress-related viability improvement in L. rhamnosus HN001, we analyzed protein synthesis in cultures subjected to different growth stages and stress conditions, using two-dimensional gel electrophoresis and N-terminal sequencing. Several proteins were up- or down regulated after either heat or osmotic shock treatments. Eleven proteins were positively identified, including the classical heat shock proteins GroEL and DnaK and the glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase, enolase, phosphoglycerate kinase, and triose phosphate isomerase, as well as tagatose 1,6-diphosphate aldolase of the tagatose pathway. The phosphocarrier protein HPr (histidine-containing proteins) was up-regulated in cultures after the log phase irrespective of the stress treatments used. The relative synthesis of an ABC transport-related protein was also up-regulated after shock treatments. Carbohydrate analysis of cytoplasmic contents showed higher levels (20 +/- 3 microg/mg of protein) in cell extracts (CFEs) derived from osmotically stressed cells than in the unstressed control (15 +/- 3 microg/mg of protein). Liquid chromatography of these crude carbohydrate extracts showed significantly different profiles. Electrospray mass spectrometry analysis of CFEs revealed, in addition to normal mono-, di-, tri-, and tetrasaccharides, the presence of saccharides modified with glycerol. PMID- 12571013 TI - Terminal restriction fragment length polymorphism data analysis for quantitative comparison of microbial communities. AB - Terminal restriction fragment length polymorphism (T-RFLP) is a culture independent method of obtaining a genetic fingerprint of the composition of a microbial community. Comparisons of the utility of different methods of (i) including peaks, (ii) computing the difference (or distance) between profiles, and (iii) performing statistical analysis were made by using replicated profiles of eubacterial communities. These samples included soil collected from three regions of the United States, soil fractions derived from three agronomic field treatments, soil samples taken from within one meter of each other in an alfalfa field, and replicate laboratory bioreactors. Cluster analysis by Ward's method and by the unweighted-pair group method using arithmetic averages (UPGMA) were compared. Ward's method was more effective at differentiating major groups within sets of profiles; UPGMA had a slightly reduced error rate in clustering of replicate profiles and was more sensitive to outliers. Most replicate profiles were clustered together when relative peak height or Hellinger-transformed peak height was used, in contrast to raw peak height. Redundancy analysis was more effective than cluster analysis at detecting differences between similar samples. Redundancy analysis using Hellinger distance was more sensitive than that using Euclidean distance between relative peak height profiles. Analysis of Jaccard distance between profiles, which considers only the presence or absence of a terminal restriction fragment, was the most sensitive in redundancy analysis, and was equally sensitive in cluster analysis, if all profiles had cumulative peak heights greater than 10,000 fluorescence units. It is concluded that T-RFLP is a sensitive method of differentiating between microbial communities when the optimal statistical method is used for the situation at hand. It is recommended that hypothesis testing be performed by redundancy analysis of Hellinger transformed data and that exploratory data analysis be performed by cluster analysis using Ward's method to find natural groups or by UPGMA to identify potential outliers. Analyses can also be based on Jaccard distance if all profiles have cumulative peak heights greater than 10,000 fluorescence units. PMID- 12571014 TI - Production of a doubly chiral compound, (4R,6R)-4-hydroxy-2,2,6 trimethylcyclohexanone, by two-step enzymatic asymmetric reduction. AB - A practical enzymatic synthesis of a doubly chiral key compound, (4R,6R)-4 hydroxy-2,2,6-trimethylcyclohexanone, starting from the readily available 2,6,6 trimethyl-2-cyclohexen-1,4-dione is described. Chirality is first introduced at the C-6 position by a stereoselective enzymatic hydrogenation of the double bond using old yellow enzyme 2 of Saccharomyces cerevisiae, expressed in Escherichia coli, as a biocatalyst. Thereafter, the carbonyl group at the C-4 position is reduced selectively and stereospecifically by levodione reductase of Corynebacterium aquaticum M-13, expressed in E. coli, to the corresponding alcohol. Commercially available glucose dehydrogenase was also used for cofactor regeneration in both steps. Using this two-step enzymatic asymmetric reduction system, 9.5 mg of (4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexanone/ml was produced almost stoichiometrically, with 94% enantiomeric excess in the presence of glucose, NAD(+), and glucose dehydrogenase. To our knowledge, this is the first report of the application of S. cerevisiae old yellow enzyme for the production of a useful compound. PMID- 12571015 TI - Desulfonation and degradation of the disulfodiphenylethercarboxylates from linear alkyldiphenyletherdisulfonate surfactants. AB - Earlier work showed that the biodegradation of a commercial linear monoalkyldiphenyletherdisulfonate surfactant as a carbon source for microbial growth leads to the quantitative formation of corresponding disulfodiphenylether carboxylates (DSDPECs), which were not degraded. alpha-Proteobacterium strain DS 1 (DSM 13023) catalyzes these reactions. These DSDPECs have now been characterized by high-pressure liquid chromatography coupled via an electrospray interface to a mass spectrometer. DSDPECs were a complex mixture of compounds which indicated catabolism via omega-oxygenation and beta-oxidation. DSDPECs were subject to quantitative desulfonation in bacterial cultures in which they served as sole sulfur sources for bacterial growth. On average, one sulfonate group per DSDPEC species was removed, and the organism responsible for this desulfonation was isolated and identified as Rhodococcus opacus ISO-5. The products were largely monosulfodiphenylether carboxylate-phenols (MSDPEC-phenols). MSDPEC phenols were subject to extensive dissimilation by bacteria from activated sludge. PMID- 12571016 TI - In situ production of exopolysaccharides during Sourdough fermentation by cereal and intestinal isolates of lactic acid bacteria. AB - EPS formed by lactobacilli in situ during sourdough fermentation may replace hydrocolloids currently used as texturizing, antistaling, or prebiotic additives in bread production. In this study, a screening of >100 strains of cereal associated and intestinal lactic acid bacteria was performed for the production of exopolysaccharides (EPS) from sucrose. Fifteen strains produced fructan, and four strains produced glucan. It was remarkable that formation of glucan and fructan was most frequently found in intestinal isolates and strains of the species Lactobacillus reuteri, Lactobacillus pontis, and Lactobacillus frumenti from type II sourdoughs. By the use of PCR primers derived from conserved amino acid sequences of bacterial levansucrase genes, it was shown that 6 of the 15 fructan-producing lactobacilli and none of 20 glucan producers or EPS-negative strains carried a levansucrase gene. In sourdough fermentations, it was determined whether those strains producing EPS in MRS medium modified as described by Stolz et al. (37) and containing 100 g of sucrose liter(-1) as the sole source of carbon also produce the same EPS from sucrose during sourdough fermentation in the presence of 12% sucrose. For all six EPS-producing strains evaluated in sourdough fermentations, in situ production of EPS at levels ranging from 0.5 to 2 g/kg of flour was demonstrated. Production of EPS from sucrose is a metabolic activity that is widespread among sourdough lactic acid bacteria. Thus, the use of these organisms in bread production may allow the replacement of additives. PMID- 12571017 TI - Growth of a Dehalococcoides-like microorganism on vinyl chloride and cis dichloroethene as electron acceptors as determined by competitive PCR. AB - A competitive PCR (cPCR) assay targeting 16S ribosomal DNA was developed to enumerate growth of a Dehalococcoides-like microorganism, bacterium VS, from a mixed culture catalyzing the reductive dehalogenation of cis-1,2-dichloroethene (cDCE) and vinyl chloride (VC), with hydrogen being used as an electron donor. The growth of bacterium VS was found to be coupled to the dehalogenation of VC and cDCE, suggesting unique metabolic capabilities. The average growth yield was (5.2 +/- 1.5) x 10(8) copies of the 16S rRNA gene/ micromol of Cl(-) (number of samples, 10), with VC being used as the electron acceptor and hydrogen as the electron donor. The maximum VC utilization rate (q) was determined to be 7.8 x 10(-10) micromol of Cl(-) (copy(-1) day(-1)), indicating a maximum growth rate of 0.4 day(-1). These average growth yield and q values agree well with values found previously for dechlorinating cultures. Decay coefficients were determined with growth (0.05 day(-1)) and no-growth (0.09 day(-1)) conditions. An important limitation of this cPCR assay was its inability to discriminate between active and inactive cells. This is an essential consideration for kinetic studies. PMID- 12571019 TI - Comparison of method 1623 and cell culture-PCR for detection of Cryptosporidium spp. in source waters. AB - Analysis of Cryptosporidium occurrence in six watersheds by method 1623 and the integrated cell culture-PCR (CC-PCR) technique provided an opportunity to evaluate these two methods. The average recovery efficiencies were 58.5% for the CC-PCR technique and 72% for method 1623, but the values were not significantly different (P = 0.06). Cryptosporidium oocysts were detected in 60 of 593 samples (10.1%) by method 1623. Infectious oocysts were detected in 22 of 560 samples (3.9%) by the CC-PCR technique. There was 87% agreement between the total numbers of samples positive as determined by method 1623 and CC-PCR for four of the sites. The other two sites had 16.3 and 24% correspondence between the methods. Infectious oocysts were detected in all of the watersheds. Overall, approximately 37% of the Cryptosporidium oocysts detected by the immunofluorescence method were viable and infectious. DNA sequence analysis of the Cryptosporidium parvum isolates detected by CC-PCR showed the presence of both the bovine and human genotypes. More than 90% of the C. parvum isolates were identified as having the bovine or bovine-like genotype. The estimates of the concentrations of infectious Cryptosporidium and the resulting daily and annual risks of infection compared well for the two methods. The results suggest that most surface water systems would require, on average, a 3-log reduction in source water Cryptosporidium levels to meet potable water goals. PMID- 12571018 TI - Phylogenetic diversity of nitrogenase (nifH) genes in deep-sea and hydrothermal vent environments of the Juan de Fuca Ridge. AB - The subseafloor microbial habitat associated with typical unsedimented mid-ocean ridge hydrothermal vent ecosystems may be limited by the availability of fixed nitrogen, inferred by the low ammonium and nitrate concentrations measured in diffuse hydrothermal fluid. Dissolved N2 gas, the largest reservoir of nitrogen in the ocean, is abundant in deep-sea and hydrothermal vent fluid. In order to test the hypothesis that biological nitrogen fixation plays an important role in nitrogen cycling in the subseafloor associated with unsedimented hydrothermal vents, degenerate PCR primers were designed to amplify the nitrogenase iron protein gene nifH from hydrothermal vent fluid. A total of 120 nifH sequences were obtained from four samples: a nitrogen-poor diffuse vent named marker 33 on Axial Volcano, sampled twice over a period of 1 year as its temperature decreased; a nitrogen-rich diffuse vent near Puffer on Endeavour Segment; and deep seawater with no detectable hydrothermal plume signal. Subseafloor nifH genes from marker 33 and Puffer are related to anaerobic clostridia and sulfate reducers. Other nifH genes unique to the vent samples include proteobacteria and divergent Archaea. All of the nifH genes from the deep-seawater sample are most closely related to the thermophilic, anaerobic archaeon Methanococcus thermolithotrophicus (77 to 83% amino acid similarity). These results provide the first genetic evidence of potential nitrogen fixers in hydrothermal vent environments and indicate that at least two sources contribute to the diverse assemblage of nifH genes detected in hydrothermal vent fluid: nifH genes from an anaerobic, hot subseafloor and nifH genes from cold, oxygenated deep seawater. PMID- 12571020 TI - Characterization of a thermostable D-stereospecific alanine amidase from Brevibacillus borstelensis BCS-1. AB - A gene encoding a new thermostable D-stereospecific alanine amidase from the thermophile Brevibacillus borstelensis BCS-1 was cloned and sequenced. The molecular mass of the purified enzyme was estimated to be 199 kDa after gel filtration chromatography and about 30 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that the enzyme could be composed of a hexamer with identical subunits. The purified enzyme exhibited strong amidase activity towards D-amino acid-containing aromatic, aliphatic, and branched amino acid amides yet exhibited no enzyme activity towards L-amino acid amides, D-amino acid-containing peptides, and NH(2)-terminally protected amino acid amides. The optimum temperature and pH for the enzyme activity were 85 degrees C and 9.0, respectively. The enzyme remained stable within a broad pH range from 7.0 to 10.0. The enzyme was inhibited by dithiothreitol, 2 mercaptoethanol, and EDTA yet was strongly activated by Co(2+) and Mn(2+). The k(cat)/K(m) for D-alaninamide was measured as 544.4 +/- 5.5 mM(-1) min(-1) at 50 degrees C with 1 mM Co(2+). PMID- 12571021 TI - Functional expression of a fungal laccase in Saccharomyces cerevisiae by directed evolution. AB - Laccase from Myceliophthora thermophila (MtL) was expressed in functional form in Saccharomyces cerevisiae. Directed evolution improved expression eightfold to the highest yet reported for a laccase in yeast (18 mg/liter). Together with a 22 fold increase in k(cat), the total activity was enhanced 170-fold. Specific activities of MtL mutants toward 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and syringaldazine indicate that substrate specificity was not changed by the introduced mutations. The most effective mutation (10-fold increase in total activity) introduced a Kex2 protease recognition site at the C-terminal processing site of the protein, adjusting the protein sequence to the different protease specificities of the heterologous host. The C terminus is shown to be important for laccase activity, since removing it by a truncation of the gene reduces activity sixfold. Mutations accumulated during nine generations of evolution for higher activity decreased enzyme stability. Screening for improved stability in one generation produced a mutant more stable than the heterologous wild type and retaining the improved activity. The molecular mass of MtL expressed in S. cerevisiae is 30% higher than that of the same enzyme expressed in M. thermophila (110 kDa versus 85 kDa). Hyperglycosylation, corresponding to a 120-monomer glycan on one N-glycosylation site, is responsible for this increase. This S. cerevisiae expression system makes MtL available for functional tailoring by directed evolution. PMID- 12571022 TI - Complete detoxification of vinyl chloride by an anaerobic enrichment culture and identification of the reductively dechlorinating population as a Dehalococcoides species. AB - A major obstacle in the implementation of the reductive dechlorination process at chloroethene-contaminated sites is the accumulation of the intermediate vinyl chloride (VC), a proven human carcinogen. To shed light on the microbiology involved in the final critical dechlorination step, a sediment-free, nonmethanogenic, VC-dechlorinating enrichment culture was derived from tetrachloroethene (PCE)-to-ethene-dechlorinating microcosms established with material from the chloroethene-contaminated Bachman Road site aquifer in Oscoda, Mich. After 40 consecutive transfers in defined, reduced mineral salts medium amended with VC, the culture lost the ability to use PCE and trichloroethene (TCE) as metabolic electron acceptors. PCE and TCE dechlorination occurred in the presence of VC, presumably in a cometabolic process. Enrichment cultures supplied with lactate or pyruvate as electron donor dechlorinated VC to ethene at rates up to 54 micromol liter(-1)day(-1), and dichloroethenes (DCEs) were dechlorinated at about 50% of this rate. The half-saturation constant (K(S)) for VC was 5.8 microM, which was about one-third lower than the concentrations determined for cis-DCE and trans-DCE. Similar VC dechlorination rates were observed at temperatures between 22 and 30 degrees C, and negligible dechlorination occurred at 4 and 35 degrees C. Reductive dechlorination in medium amended with ampicillin was strictly dependent on H(2) as electron donor. VC-dechlorinating cultures consumed H(2) to threshold concentrations of 0.12 ppm by volume. 16S rRNA gene based tools identified a Dehalococcoides population, and Dehalococcoides-targeted quantitative real-time PCR confirmed VC-dependent growth of this population. These findings demonstrate that Dehalococcoides populations exist that use DCEs and VC but not PCE or TCE as metabolic electron acceptors. PMID- 12571023 TI - Comparative genetic diversity of the narG, nosZ, and 16S rRNA genes in fluorescent pseudomonads. AB - The diversity of the membrane-bound nitrate reductase (narG) and nitrous oxide reductase (nosZ) genes in fluorescent pseudomonads isolated from soil and rhizosphere environments was characterized together with that of the 16S rRNA gene by a PCR-restriction fragment length polymorphism assay. Fragments of 1,008 bp and 1,433 bp were amplified via PCR with primers specific for the narG and nosZ genes, respectively. The presence of the narG and nosZ genes in the bacterial strains was confirmed by hybridization of the genomic DNA and the PCR products with the corresponding probes. The ability of the strains to either reduce nitrate or totally dissimilate nitrogen was assessed. Overall, there was a good correspondence between the reductase activities and the presence of the corresponding genes. Distribution in the different ribotypes of strains harboring both the narG and nosZ genes and of strains missing both genes suggests that these two groups of strains had different evolutionary histories. Both dissimilatory genes showed high polymorphism, with similarity indexes (Jaccard) of between 0.04 and 0.8, whereas those of the 16S rRNA gene only varied from 0.77 to 0.99. No correlation between the similarity indexes of 16S rRNA and dissimilatory genes was seen, suggesting that the evolution rates of ribosomal and functional genes differ. Pairwise comparison of similarity indexes of the narG and nosZ genes led to the delineation of two types of strains. Within the first type, the similarity indexes of both genes varied in the same range, suggesting that these two genes have followed a similar evolution. Within the second type of strain, the range of variations was higher for the nosZ than for the narG gene, suggesting that these genes have had a different evolutionary rate. PMID- 12571024 TI - Three transporters mediate uptake of glycine betaine and carnitine by Listeria monocytogenes in response to hyperosmotic stress. AB - The uptake and accumulation of the potent osmolytes glycine betaine and carnitine enable the food-borne pathogen Listeria monocytogenes to proliferate in environments of elevated osmotic stress, often rendering salt-based food preservation inadequate. To date, three osmolyte transport systems are known to operate in L. monocytogenes: glycine betaine porter I (BetL), glycine betaine porter II (Gbu), and a carnitine transporter OpuC. We investigated the specificity of each transporter towards each osmolyte by creating mutant derivatives of L. monocytogenes 10403S that possess each of the transporters in isolation. Kinetic and steady-state osmolyte accumulation data together with growth rate experiments demonstrated that osmotically activated glycine betaine transport is readily and effectively mediated by Gbu and BetL and to a lesser extent by OpuC. Osmotically stimulated carnitine transport was demonstrated for OpuC and Gbu regardless of the nature of stressing salt. BetL can mediate weak carnitine uptake in response to NaCl stress but not KCl stress. No other transporter in L. monocytogenes 10403S appears to be involved in osmotically stimulated transport of either osmolyte, since a triple mutant strain yielded neither transport nor accumulation of glycine betaine or carnitine and could not be rescued by either osmolyte when grown under elevated osmotic stress. PMID- 12571025 TI - Cloning, sequencing, and expression of the chitinase gene chiA74 from Bacillus thuringiensis. AB - The endochitinase gene chiA74 from Bacillus thuringiensis serovar kenyae strain LBIT-82 was cloned in Escherichia coli DH5 alpha F'. A sequence of 676 amino acids was deduced when the gene was completely sequenced. A molecular mass of 74 kDa was estimated for the preprotein, which includes a putative 4-kDa signal sequence located at the N terminus. The deduced amino acid sequence showed high degree of identity with other chitinases such as ChiB from Bacillus cereus (98%) and ChiA71 from Bacillus thuringiensis serovar pakistani (70%). Additionally, ChiA74 showed a modular structure comprised of three domains: a catalytic domain, a fibronectin-like domain, and a chitin-binding domain. All three domains showed conserved sequences when compared to other bacterial chitinase sequences. A ca. 70-kDa mature protein expressed by the cloned gene was detected in zymograms, comigrating with a chitinase produced by the LBIT-82 wild-type strain. ChiA74 is active within a wide pH range (4 to 9), although a bimodal activity was shown at pH 4.79 and 6.34. The optimal temperature was estimated at 57.2 degrees C when tested at pH 6. The potential use of ChiA74 as a synergistic agent, along with the B. thuringiensis insecticidal Cry proteins, is discussed. PMID- 12571026 TI - Depth distribution of microbial diversity in Mono Lake, a meromictic soda lake in California. AB - We analyzed the variation with depth in the composition of members of the domain Bacteria in samples from alkaline, hypersaline, and currently meromictic Mono Lake in California. DNA samples were collected from the mixolimnion (2 m), the base of the oxycline (17.5 m), the upper chemocline (23 m), and the monimolimnion (35 m). Composition was assessed by sequencing randomly selected cloned fragments of 16S rRNA genes retrieved from the DNA samples. Most of the 212 sequences retrieved from the samples fell into five major lineages of the domain Bacteria: alpha- and gamma-Proteobacteria (6 and 10%, respectively), Cytophaga-Flexibacter Bacteroides (19%), high-G+C-content gram-positive organisms (Actinobacteria; 25%), and low-G+C-content gram-positive organisms (Bacillus and Clostridium; 19%). Twelve percent were identified as chloroplasts. The remaining 9% represented beta- and delta-Proteobacteria, Verrucomicrobiales, and candidate divisions. Mixolimnion and oxycline samples had low microbial diversity, with only 9 and 12 distinct phylotypes, respectively, whereas chemocline and monimolimnion samples were more diverse, containing 27 and 25 phylotypes, respectively. The compositions of microbial assemblages from the mixolimnion and oxycline were not significantly different from each other (P = 0.314 and 0.877), but they were significantly different from those of chemocline and monimolimnion assemblages (P < 0.001), and the compositions of chemocline and monimolimnion assemblages were not significantly different from each other (P = 0.006 and 0.124). The populations of sequences retrieved from the mixolimnion and oxycline samples were dominated by sequences related to high-G+C-content gram-positive bacteria (49 and 63%, respectively) distributed in only three distinct phylotypes, while the population of sequences retrieved from the monimolimnion sample was dominated (52%) by sequences related to low-G+C-content gram-positive bacteria distributed in 12 distinct phylotypes. Twelve and 28% of the sequences retrieved from the chemocline sample were also found in the mixolimnion and monimolimnion samples, respectively. None of the sequences retrieved from the monimolimnion sample were found in the mixolimnion or oxycline samples. Elevated diversity in anoxic bottom water samples relative to oxic surface water samples suggests a greater opportunity for niche differentiation in bottom versus surface waters of this lake. PMID- 12571027 TI - Seasonal changes in fungal production and biomass on standing dead Scirpus lacustris litter in a northern prairie wetland. AB - Decaying macrophytes are an important source of carbon and nutrients in fungal and bacterial communities of northern prairie wetlands. Dead macrophytes do not collapse into the water column immediately after death, and decomposition by fungi and bacteria begins while the plants are standing. The seasonal variations in fungal biomass and production on Scirpus lacustris stems, both above and below water, were measured to assess which environmental factors were dominant in affecting these variations in a typical prairie wetland. Fungal biomass and production were measured from early May to November, just prior to freeze-up. Fungal decomposition began and was greatest in the spring despite low water temperatures. The fungal production, as measured by the incorporation of [1 (14)C]acetate into ergosterol, ranged from 1.8 to 376 microg of C g of ash-free dry mass (AFDM)(-1) day(-1), and the biomass, as estimated by using ergosterol, ranged from nondetectable to 5.8 mg of C g of AFDM(-1). There was no significant difference in biomass or production between aerial and submerged portions of Scirpus stems. The water temperature was correlated with fungal production (r = 0.7, P < 0.005) for aerial stem pieces but not for submerged pieces. However, in laboratory experiments water temperature had a measurable effect on both biomass and production in submerged stem pieces. Changes in fungal biomass and productivity on freshly cut green Scirpus stems decaying in the water either exposed to natural solar radiation or protected from UV radiation were monitored over the summer. There was no significant difference in either fungal biomass (P = 0.76) or production (P = 0.96) between the two light treatments. The fungal biomass and rates of production were within the lower range of the values reported elsewhere, probably as a result of the colder climate and perhaps the lower lability of Scirpus stems compared to the labilities of the leaves and different macrophytes examined in other studies performed at lower latitudes. PMID- 12571028 TI - Bacteriocin detection from whole bacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry. AB - Class I bacteriocins (lantibiotics) and class II bacteriocins are antimicrobial peptides secreted by gram-positive bacteria. Using two lantibiotics, lacticin 481 and nisin, and the class II bacteriocin coagulin, we showed that bacteriocins can be detected without any purification from whole producer bacteria grown on plates by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). When we compared the results of MALDI-TOF-MS performed with samples of whole cells and with samples of crude supernatants of liquid cultures, the former samples led to more efficient bacteriocin detection and required less handling. Nisin and lacticin 481 were both detected from a mixture of their producer strains, but such a mixture can yield additional signals. We used this method to determine the masses of two lacticin 481 variants, which confirmed at the peptide level the effect of mutations in the corresponding structural gene. PMID- 12571029 TI - Comparison of Shiga toxin production by hemolytic-uremic syndrome-associated and bovine-associated Shiga toxin-producing Escherichia coli isolates. AB - There is considerable diversity among Shiga toxin (Stx)-producing Escherichia coli (STEC) bacteria, and only a subset of these organisms are thought to be human pathogens. The characteristics that distinguish STEC bacteria that give rise to human disease are not well understood. Stxs, the principal virulence determinants of STEC, are thought to account for hemolytic-uremic syndrome (HUS), a severe clinical consequence of STEC infection. Stxs are typically bacteriophage encoded, and their production has been shown to be enhanced by prophage-inducing agents such as mitomycin C in a limited number of clinical STEC isolates. Low iron concentrations also enhance Stx production by some clinical isolates; however, little is known regarding whether and to what extent these stimuli regulate Stx production by STEC associated with cattle, the principal environmental reservoir of STEC. In this study, we investigated whether toxin production differed between HUS- and bovine-associated STEC strains. Basal production of Stx by HUS-associated STEC exceeded that of bovine-associated STEC. In addition, following mitomycin C treatment, Stx2 production by HUS-associated STEC was significantly greater than that by bovine-associated STEC. Unexpectedly, mitomycin C treatment had a minimal effect on Stx1 production by both HUS- and bovine-associated STEC. However, Stx1 production was induced by growth in low iron medium, and induction was more marked for HUS-associated STEC than for bovine-associated STEC. These observations reveal that disease-associated and bovine-associated STEC bacteria differ in their basal and inducible Stx production characteristics. PMID- 12571030 TI - Symbiotic and genetic diversity of Rhizobium galegae isolates collected from the Galega orientalis gene center in the Caucasus. AB - This paper explores the relationship between the genetic diversity of rhizobia and the morphological diversity of their plant hosts. Rhizobium galegae strains were isolated from nodules of wild Galega orientalis and Galega officinalis in the Caucasus, the center of origin for G. orientalis. All 101 isolates were characterized by genomic amplified fragment length polymorphism fingerprinting and by PCR-restriction fragment length polymorphism (RFLP) of the rRNA intergenic spacer and of five parts of the symbiotic region adjacent to nod box sequences. By all criteria, the R. galegae bv. officinalis and R. galegae bv. orientalis strains form distinct clusters. The nod box regions are highly conserved among strains belonging to each of the two biovars but differ structurally to various degrees between the biovars. The findings suggest varying evolutionary pressures in different parts of the symbiotic genome of closely related R. galegae biovars. Sixteen R. galegae bv. orientalis strains harbored copies of the same insertion sequence element; all were isolated from a particular site and belonged to a limited range of chromosomal genotypes. In all analyses, the Caucasian R. galegae bv. orientalis strains were more diverse than R. galegae bv. officinalis strains, in accordance with the gene center theory. PMID- 12571031 TI - Molecular analyses of Salmonella enterica isolates from fish feed factories and fish feed ingredients. AB - Isolates of the most commonly observed salmonella serovars in Norwegian fish feed factories from 1998 to 2000 (Salmonella enterica serovar Agona, S. enterica serovar Montevideo, S. enterica serovar Senftenberg, and S. enterica serovar Kentucky) were studied by pulsed-field gel electrophoresis (PFGE) and plasmid profile analysis and compared to isolates of the same serovars from fish feed ingredients, humans, and other sources (a total of 112 isolates). Within each serovar, a variety of distinct PFGE types (with similarity levels less than 90%) were observed in the feed ingredients and other sources, while only two distinct types of each serovar were identified in the factories. The combined results of PFGE and plasmid analyses showed that each factory harbored only a few S. enterica clones. Some of these clones persisted for at least 3 years in the factories, indicating that there was long-lasting contamination probably due to inadequate decontamination procedures. PMID- 12571032 TI - Detection of Listeria monocytogenes from a model food by fluorescence resonance energy transfer-based PCR with an asymmetric fluorogenic probe set. AB - It has been shown that fluorescence resonance energy transfer (FRET)-based PCR, including the TaqMan assay and molecular beacons, has potential for rapid detection of pathogens. In these promising real-time detection assays a single internal oligonucleotide probe labeled on both the 5' (reporter) and 3' (quencher) ends is used for selective generation of fluorescence. In this paper, we describe the use of a previously reported novel probe design for FRET-based PCR detection of Listeria monocytogenes in pure culture and in a model food commodity. In the assay described here an asymmetric probe set is used; this probe set consists of a long 5' fluorescein-labeled reporter probe and a short, complementary 3' DABCYL-labeled quencher oligonucleotide, which are used in a 5' nuclease amplification and detection assay. By using the listeriolysin O (hly) and p60 (iap) genes as amplification targets, the performance of two primer-probe sets in amplification and subsequent detection of target DNA was evaluated. In studies performed with pure cultures of L. monocytogenes, the PCR profiles indicated that the relative change in fluorescence intensity was correlated with both the initial number of cells and the accumulation of specific amplicons for both hly and iap gene fragments. Experiments were also done to determine the applicability of the method to the detection of L. monocytogenes by targeting hly DNA and its short-lived mRNA product in a model food commodity. Twenty-five milliliter samples of reconstituted nonfat dry milk (NFDM) were seeded with L. monocytogenes and processed to concentrate the bacteria by centrifugation, and this was followed by nucleic acid extraction and amplification with hly-specific primers. Endpoint detection of PCR and reverse transcription-PCR amplicons could be achieved at inoculum levels of 10(3) and 10(4) CFU of L. monocytogenes/25 ml of NFDM, respectively. This study demonstrated that this asymmetric FRET-based amplification and detection protocol provides an alternative approach for endpoint detection of nucleic acid amplification products as applied to detection of pathogens in a model food. PMID- 12571033 TI - Geographical variation in ribotype profiles of Escherichia coli isolates from humans, swine, poultry, beef, and dairy cattle in Florida. AB - Waters impacted by fecal pollution can exact high risks to human health and can result in financial losses due to closures of water systems used for recreation and for harvesting seafood. Identifying the sources of fecal pollution in water is paramount in assessing the potential human health risks involved as well as in assessing necessary remedial action. Recently, various researchers have used the ribotyping method to identify sources of bacterial indicators (Escherichia coli and enterococci) in environmental waters. While these studies have identified genotypic differences between human- and animal-derived indicators that are capable of differentiating organisms isolated from humans and various animal hosts, most have focused on organisms collected from a confined geographic area and have not addressed the question of whether these ribotype profiles are watershed specific or if they can be applied universally to organisms from other geographic locations. In this study, E. coli isolates were obtained from humans, beef cattle, dairy cattle, swine, and poultry from locations in northern, central, and southern Florida and were subjected to ribotyping analysis. The intent was to determine (i) if ribotype profiles are capable of discriminating the source of E. coli at the host species level and (ii) if the resulting fingerprints are uniform over an extended geographic area or if they can be applied only to a specific watershed. Our research indicated that, using a single restriction enzyme (HindIII), the ribotyping procedure is not capable of differentiating E. coli isolates from the different animal species sampled in this study. Results indicate, however, that this procedure can still be used effectively to differentiate E. coli as being either human or animal derived when applied to organisms isolated from a large geographic region. PMID- 12571034 TI - A combined model to predict the functionality of the bacteriocin-producing Lactobacillus sakei strain CTC 494. AB - The use of bacteriocin-producing lactic acid bacteria for improved food fermentation processes seems promising. However, lack of fundamental knowledge about the functionality of bacteriocin-producing strains under food fermentation conditions hampers their industrial use. Predictive microbiology or a mathematical estimation of microbial behavior in food ecosystems may help to overcome this problem. In this study, a combined model was developed that was able to estimate, from a given initial situation of temperature, pH, and nutrient availability, the growth and self-inhibition dynamics of a bacteriocin-producing Lactobacillus sakei CTC 494 culture in (modified) MRS broth. Moreover, the drop in pH induced by lactic acid production and the bacteriocin activity toward Listeria as an indicator organism were modeled. Self-inhibition was due to the depletion of nutrients as well as to the production of lactic acid. Lactic acid production resulted in a pH drop, an accumulation of toxic undissociated lactic acid molecules, and a shift in the dissociation degree of the growth-inhibiting buffer components. The model was validated experimentally. PMID- 12571035 TI - Small, acid-soluble proteins as biomarkers in mass spectrometry analysis of Bacillus spores. AB - The use of 1 N HCl for extraction of small, acid-soluble proteins (SASP) from different Bacillus spore species was examined. The extracts were analyzed by high performance liquid chromatography and matrix-assisted laser desorption mass spectrometry and were found to be both qualitatively and quantitatively superior to extraction by acetonitrile-5% trifluoroacetic acid (70:30, vol/vol). Both major and minor alpha/beta- and gamma-type SASP were characterized by their molecular masses or tryptic peptide maps and by searches of both protein and unannotated genome databases. For all but 1 pair (B. cereus T and B. thuringiensis subsp. Kurstaki) among the 11 variants studied the suites of SASP masses are distinctive, consistent with the use of these proteins as potential biomarkers for spore identification by mass spectrometry. PMID- 12571037 TI - Differential inactivation of seed exudate stimulation of Pythium ultimum sporangium germination by Enterobacter cloacae influences biological control efficacy on different plant species. AB - This study was initiated to understand whether differential biological control efficacy of Enterobacter cloacae on various plant species is due to differences in the ability of E. cloacae to inactivate the stimulatory activity of seed exudates to Pythium ultimum sporangium germination. In biological control assays, E. cloacae was effective in controlling Pythium damping-off when placed on the seeds of carrot, cotton, cucumber, lettuce, radish, tomato, and wheat but failed to protect corn and pea from damping-off. Seeds from plants such as corn and pea had high rates of exudation, whereas cotton and cucumber seeds had much lower rates of exudation. Patterns of seed exudation and the release of P. ultimum sporangium germination stimulants varied among the plants tested. Seed exudates of plants such as carrot, corn, lettuce, pea, radish, and wheat were generally more stimulatory to P. ultimum than were the exudates of cotton, cucumber, sunflower, and tomato. However, this was not directly related to the ability of E. cloacae to inactivate the stimulatory activity of the exudate and reduce P. ultimum sporangium germination. In the spermosphere, E. cloacae readily reduced the stimulatory activity of seed exudates from all plant species except corn and pea. Our data have shown that the inability of E. cloacae to protect corn and pea seeds from Pythium damping-off is directly related to its ability to inactivate the stimulatory activity of seed exudates. On all other plants tested, E. cloacae was effective in suppressing damping-off and inactivating the stimulatory activity of seed exudates. PMID- 12571036 TI - A single mutation in the activation site of bovine trypsinogen enhances its accumulation in the fermentation broth of the yeast Pichia pastoris. AB - We produced bovine trypsinogen in the yeast Pichia pastoris. Little or no trypsinogen was detected when the gene with its native leader sequence was expressed under the control of the strong aox1 promoter, suggesting that expression of the wild-type bovine trypsinogen was toxic to the cells. We altered the trypsinogen native propeptide sequence by replacing the lysine at position 6 with an aspartic acid, thus destroying the site in the propeptide cleaved by enterokinase and by trypsin. This mutant accumulated up to 10 mg of trypsinogen per liter in shake flask cultures and about 40 mg/liter in 6-liter fermentors. Trypsinogen could be activated in vitro with a dipeptidyl-aminopeptidase, which selectively removed the modified trypsinogen propeptide; the resulting trypsin was fully active and showed evidence of glycosylation. Thus, we have developed a novel protein production scheme that can be used for the expression of proteins, such as proteases, that are deleterious to the producing organism. This system relies on the expression of a zymogen that cannot be activated in vivo coupled with its in vitro purification and activation. PMID- 12571038 TI - Genetic manipulation of Lactococcus lactis by using targeted group II introns: generation of stable insertions without selection. AB - Despite their commercial importance, there are relatively few facile methods for genomic manipulation of the lactic acid bacteria. Here, the lactococcal group II intron, Ll.ltrB, was targeted to insert efficiently into genes encoding malate decarboxylase (mleS) and tetracycline resistance (tetM) within the Lactococcus lactis genome. Integrants were readily identified and maintained in the absence of a selectable marker. Since splicing of the Ll.ltrB intron depends on the intron-encoded protein, targeted invasion with an intron lacking the intron open reading frame disrupted TetM and MleS function, and MleS activity could be partially restored by expressing the intron-encoded protein in trans. Restoration of splicing from intron variants lacking the intron-encoded protein illustrates how targeted group II introns could be used for conditional expression of any gene. Furthermore, the modified Ll.ltrB intron was used to separately deliver a phage resistance gene (abiD) and a tetracycline resistance marker (tetM) into mleS, without the need for selection to drive the integration or to maintain the integrant. Our findings demonstrate the utility of targeted group II introns as a potential food-grade mechanism for delivery of industrially important traits into the genomes of lactococci. PMID- 12571039 TI - Engineering of carbon distribution between glycolysis and sugar nucleotide biosynthesis in Lactococcus lactis. AB - We describe the effects of modulating the activities of glucokinase, phosphofructokinase, and phosphoglucomutase on the branching point between sugar degradation and the biosynthesis of sugar nucleotides involved in the production of exopolysaccharide biosynthesis by Lactococcus lactis. This was realized by using a described isogenic L. lactis mutant with reduced enzyme activities or by controlled expression of the well-characterized genes for phosphoglucomutase or glucokinase from Escherichia coli or Bacillus subtilis, respectively. The role of decreased metabolic flux was studied in L. lactis strains with decreased phosphofructokinase activities. The concomitant reduction of the activities of phosphofructokinase and other enzymes encoded by the las operon (lactate dehydrogenase and pyruvate kinase) resulted in significant changes in the concentrations of sugar-phosphates. In contrast, a >25-fold overproduction of glucokinase resulted in 7-fold-increased fructose-6-phosphate levels and 2-fold reduced glucose-1-phosphate and glucose-6-phosphate levels. However, these increased sugar-phosphate concentrations did not affect the levels of sugar nucleotides. Finally, an approximately 100-fold overproduction of phosphoglucomutase resulted in 5-fold-increased levels of both UDP-glucose and UDP-galactose. While the increased concentrations of sugar-phosphates or sugar nucleotides did not significantly affect the production of exopolysaccharides, they demonstrate the metabolic flexibility of L. lactis. PMID- 12571040 TI - Effects of alternative dietary substrates on competition between human colonic bacteria in an anaerobic fermentor system. AB - Duplicate anaerobic fermentor systems were used to examine changes in a community of human fecal bacteria supplied with different carbohydrate energy sources. A panel of group-specific fluorescent in situ hybridization probes targeting 16S rRNA sequences revealed that the fermentors supported growth of a greater proportion of Bacteroides and a lower proportion of gram-positive anaerobes related to Faecalibacterium prausnitzii, Ruminococcus flavefaciens-Ruminococcus bromii, Eubacterium rectale-Clostridium coccoides, and Eubacterium cylindroides than the proportions in the starting fecal inoculum. Nevertheless, certain substrates, such as dahlia inulin, caused a pronounced increase in the number of bacteria related to R. flavefaciens-R. bromii and E. cylindroides. The ability of three strictly anaerobic, gram-positive bacteria to compete with the complete human fecal flora was tested in the same experiment by using selective plating to enumerate the introduced strains. The Roseburia-related strain A2-183(F) was able to grow on all substrates despite the fact that it was unable to utilize complex carbohydrates in pure culture, and it was assumed that this organism survived by cross-feeding. In contrast, Roseburia intestinalis L1-82(R) and Eubacterium sp. strain A2-194(R) survived less well despite the fact that they were able to utilize polysaccharides in pure culture, except that A2-194(R) was stimulated 100 fold by inulin. These results suggest that many low-G+C-content gram-positive obligate anaerobes may be selected against during in vitro incubation, although several groups were stimulated by inulin. Thus, considerable caution is necessary when workers attempt to predict the in vivo effects of probiotics and prebiotics from their effects in vitro. PMID- 12571041 TI - High-performance liquid chromatography analyses of pyoverdin siderophores differentiate among phytopathogenic fluorescent Pseudomonas Species. AB - The relationship of pyoverdins produced by 41 pathovars of Pseudomonas syringae and by phytopathogenic Pseudomonas species was investigated. A high-performance liquid chromatography method for analyzing the culture medium proved to be superior to isoelectric focusing for detecting pyoverdin production, for differentiating slightly different pyoverdins, and for differentiating atypical from typical Fe(III)-chelated pyoverdins. Nonfluorescent strains were found in Pseudomonas amygdali, Pseudomonas meliae, Pseudomonas fuscovaginae, and P. syringae. Pseudomonas agarici and Pseudomonas marginalis produced typical pyoverdins. Among the arginine dihydrolase-negative fluorescent Pseudomonas species, spectral, amino acid, and mass spectrometry analyses underscored for the first time the clear similarities among the pyoverdins produced by related species. Within this group, the oxidase-negative species Pseudomonas viridiflava and Pseudomonas ficuserectae and the pathovars of P. syringae produced the same atypical pyoverdin, whereas the oxidase-positive species Pseudomonas cichorii produced a similar atypical pyoverdin that contained a glycine instead of a serine. The more distantly related species Pseudomonas asplenii and Pseudomonas fuscovaginae both produced a less similar atypical pyoverdin. The spectral characteristics of Fe(III)-chelated atypical pyoverdins at pH 7.0 were related to the presence of two beta-hydroxyaspartic acids as iron ligands, whereas in typical pyoverdins one of the ligands is always ornithine based. The peptide chain influenced the chelation of iron more in atypical pyoverdins. Our results demonstrated that there is relative pyoverdin conservation in the amino acids involved in iron chelation and that there is faster evolution of the other amino acids, highlighting the usefulness of pyoverdins in systematics and in identification. PMID- 12571042 TI - Monitoring the production of aflatoxin B1 in wheat by measuring the concentration of nor-1 mRNA. AB - A real-time reverse transcription-PCR system has been used to monitor the expression of an aflatoxin biosynthetic gene of Aspergillus flavus in wheat. Therefore, total RNA was isolated from infected wheat samples, reverse transcribed and subjected to real-time PCR. In parallel all samples were analyzed by high-pressure liquid chromatography for aflatoxin B(1) production. The primer probe system of the real-time PCR was targeted against nor-1, a gene of the aflatoxin biosynthetic pathway. By application of this method the nor-1 transcription was quantified during the course of incubation. After 4 days of incubation nor-1 mRNA could be detected for the first time. The amount of nor-1 mRNA increased rapidly, and the maximum was achieved after 6 days. Then, starting very slowly, the mRNA was degraded until day 8, and this was followed by a very fast degradation, reaching nondetectable levels at days 9 and 10. First traces of aflatoxin B(1)could be detected between the 5th and 6th day of incubation. The aflatoxin concentration reached its maximum after 9 days of incubation and remained constant for the whole period of observation. To ensure that differences in the nor-1 mRNA concentration were due to different expression levels, the expression of the constitutively expressed beta-tubulin gene (benA56) has also been monitored. The expression of benA56 remained constant during the whole incubation time. As a parameter for fungal growth, the number of nor-1 gene copies was determined during the course of incubation. The numbers of nor-1 gene copies increased at the beginning of the incubation and reached a plateau at day 5. They correlate well with the viable counts albeit at a higher level. PMID- 12571043 TI - Oligonucleotide microarray for the study of functional gene diversity in the nitrogen cycle in the environment. AB - The analysis of functional diversity and its dynamics in the environment is essential for understanding the microbial ecology and biogeochemistry of aquatic systems. Here we describe the development and optimization of a DNA microarray method for the detection and quantification of functional genes in the environment and report on their preliminary application to the study of the denitrification gene nirS in the Choptank River-Chesapeake Bay system. Intergenic and intragenic resolution constraints were determined by an oligonucleotide (70 mer) microarray approach. Complete signal separation was achieved when comparing unrelated genes within the nitrogen cycle (amoA, nifH, nirK, and nirS) and detecting different variants of the same gene, nirK, corresponding to organisms with two different physiological modes, ammonia oxidizers and denitrifying halobenzoate degraders. The limits of intragenic resolution were investigated with a microarray containing 64 nirS sequences comprising 14 cultured organisms and 50 clones obtained from the Choptank River in Maryland. The nirS oligonucleotides covered a range of sequence identities from approximately 40 to 100%. The threshold values for specificity were determined to be 87% sequence identity and a target-to-probe perfect match-to-mismatch binding free-energy ratio of 0.56. The lower detection limit was 10 pg of DNA (equivalent to approximately 10(7) copies) per target per microarray. Hybridization patterns on the microarray differed between sediment samples from two stations in the Choptank River, implying important differences in the composition of the denitirifer community along an environmental gradient of salinity, inorganic nitrogen, and dissolved organic carbon. This work establishes a useful set of design constraints (independent of the target gene) for the implementation of functional gene microarrays for environmental applications. PMID- 12571044 TI - Incidence of enteric viruses in groundwater from household wells in Wisconsin. AB - Recent studies on the contamination of groundwater with human enteric viruses have focused on public water systems, whereas little is known about the occurrence of viruses in private household wells. The objective of the present study was to estimate the incidence of viruses in Wisconsin household wells located near septage land application sites or in rural subdivisions served by septic systems. Fifty wells in seven hydrogeologic districts were sampled four times over a year, once each season. Reverse transcriptase PCR (RT-PCR), followed by Southern hybridization, was used to detect enteroviruses, rotavirus, hepatitis A virus (HAV), and Norwalk-like viruses (NLVs). In addition, cell culture was used to detect culturable enteroviruses. Companion water samples were collected for total coliforms, Escherichia coli, fecal enterococci, F-specific RNA coliphages, nitrate, and chloride analyses. Among the 50 wells, four (8%) were positive for viruses by RT-PCR. Three wells were positive for HAV, and the fourth well was positive for both rotavirus and NLV in one sample and an enterovirus in another sample. Contamination was transient, since none of the wells was virus positive for two sequential samples. Culturable enteroviruses were not detected in any of the wells. Water quality indicators were not statistically associated with virus occurrence, although some concordance was noted for chloride. The present study is the first in the United States to systematically monitor private household wells for virus contamination and, combined with data for public wells, provides further insight on the extent of groundwater contamination with human enteric viruses. PMID- 12571045 TI - Specific detection of Arcobacter and Campylobacter strains in water and sewage by PCR and fluorescent in situ hybridization. AB - The aim of this study was to evaluate PCR and fluorescent in situ hybridization (FISH) techniques for detecting Arcobacter and Campylobacter strains in river water and wastewater samples. Both 16S and 23S rRNA sequence data were used to design specific primers and oligonucleotide probes for PCR and FISH analyses, respectively. In order to assess the suitability of the methods, the assays were performed on naturally and artificially contaminated samples and compared with the isolation of cells on selective media. The detection range of PCR and FISH assays varied between 1 cell/ml (after enrichment) to 10(3) cells/ml (without enrichment). According to our results, both rRNA-based techniques have the potential to be used as quick and sensitive methods for detection of campylobacters in environmental samples. PMID- 12571046 TI - Involvement of calcium/calmodulin signaling in cercosporin toxin biosynthesis by Cercospora nicotianae. AB - Cercosporin is a non-host-selective, perylenequinone toxin produced by many phytopathogenic Cercospora species. The involvement of Ca(2+)/calmodulin (CaM) signaling in cercosporin biosynthesis was investigated by using pharmacological inhibitors. The results suggest that maintaining endogenous Ca(2+) homeostasis is required for cercosporin biosynthesis in Cercospora nicotianae. The addition of excess Ca(2+) to the medium slightly increased fungal growth but resulted in a reduction in cercosporin production. The addition of Ca(2+) chelators [EGTA and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid] also reduced cercosporin production. Ca(2+) channel blockers exhibited a strong inhibition of cercosporin production only at higher concentrations (>2 mM). Cercosporin production was reduced greatly by Ca(2+) ionophores (A23187 and ionomycin) and internal Ca(2+) blocker [3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester]. Phospholipase C inhibitors (lithium, U73122, and neomycin) led to a concentration-dependent inhibition of cercosporin biosynthesis. Furthermore, the addition of CaM inhibitors (compound 48/80, trifluoperazine, W-7, and chlorpromazine) also markedly reduced cercosporin production. In contrast to W-7, W-5, with less specificity for CaM, led to only minor inhibition of cercosporin production. The inhibitory effects of Ca(2+)/CaM inhibitors were partially or completely reversed by the addition of external Ca(2+). As assessed with Fluo 3/AM (a fluorescent Ca(2+) indicator), the Ca(2+) content in the cytoplasm decreased significantly when fungal cultures were grown in a medium containing Ca(2+)/CaM antagonists, confirming the specificity of those Ca(2+)/CaM antagonists in C. nicotianae. Taken together, the results suggest that Ca(2+)/CaM signal transduction may play a pivotal role in cercosporin biosynthesis in C. nicotianae. PMID- 12571047 TI - Use of a promoter trap to identify Bacillus cereus genes regulated by tomato seed exudate and a rhizosphere resident, Pseudomonas aureofaciens. AB - The goal of this study was to identify genes in Bacillus cereus, a bacterium commonly associated with plant seeds and roots, that are affected by compounds originating from a host plant, tomato, or another rhizosphere resident, Pseudomonas aureofaciens. We constructed a B. cereus chromosomal DNA library in a promoter-trap plasmid, pAD123, which contains a promoterless version of the green fluorescent protein (GFP) gene, gfpmut3a. The library was screened by using fluorescence-activated cell sorting for clones showing a change in GFP expression in response to either tomato seed exudate or culture supernatant of P. aureofaciens strain 30-84. We identified two clones carrying genes that were induced by the presence of tomato seed exudate and nine clones carrying genes that were repressed by P. aureofaciens culture supernatant. A clone chosen for further study contained an open reading frame, designated lipA, that encodes a deduced protein with a lipoprotein signal peptide sequence similar to lipoproteins in B. subtilis. Expression of gusA under control of the lipA promoter increased twofold when cells were exposed to tomato seed exudate and in a concentration-dependent manner when exposed to a mixture of amino acids. When the wild type and a 10-fold excess of a lipA mutant were applied together to tomato seeds, 2 days after planting, the wild type displayed medium-dependent culturability, whereas the lipA mutant was unaffected. This study demonstrates the power of a promoter trap to identify genes in a gram-positive bacterium that are regulated by the biotic environment and resulted in the discovery of lipA, a plant-regulated gene in B. cereus. PMID- 12571048 TI - Biotin limitation in Sinorhizobium meliloti strain 1021 alters transcription and translation. AB - Most Sinorhizobium meliloti strains lack several key genes involved in microbial biotin biosynthesis, and it is assumed that this may be a special adaptation which allows the microbe to down-regulate metabolic activities in the absence of a host plant. To further explore this hypothesis, we employed two different strategies. (i) Searches of the S. meliloti genome database in combination with the construction of nine different gusA reporter fusions identified three genes involved in a biotin starvation response in this microbe. A gene coding for a protein-methyl carboxyl transferase (pcm) exhibited 13.6-fold-higher transcription under biotin-limiting conditions than cells grown in the presence of 40 nM biotin. Consistent with this observation, biotin-limiting conditions resulted in a significantly decreased survival of pcm mutant cells compared to parental cells or cells grown in the presence of 40 nM biotin. Further studies indicated that the autoinducer synthase gene, sinI, was transcribed at a 4.5-fold higher level in early stationary phase in biotin-starved cells than in biotin supplemented cells. Lastly, we observed that open reading frame smc02283, which codes for a putative copper resistance protein (CopC), was 21-fold down-regulated in response to biotin starvation. (ii) In a second approach, proteome analysis identified 10 proteins which were significantly down-regulated under the biotin limiting conditions. Among the proteins identified by using matrix-assisted laser desorption ionization-time of flight mass spectrometry were the pi subunit of the RNA polymerase and the 50S ribosomal protein L7/L12 (L8) subunit, indicating that biotin-limiting conditions generally affect transcription and translation in S. meliloti. PMID- 12571049 TI - Development of Sinorhizobium meliloti pilot macroarrays for transcriptome analysis. AB - In order to prepare for whole-genome expression analysis in Sinorhizobium meliloti, pilot DNA macroarrays were designed for 34 genes of known regulation. The experimental parameters assessed were the length of the PCR products, the influence of a tag at the 5' end of the primers, and the method of RNA labeling. Variance and principal-component analysis showed that the most important nonbiological parameter was the labeling method. The sizes of PCR products were also found to be important, whereas the influence of 5' tags was minimal. The variability between replicated spots on a membrane was found to be low. These experimental procedures were validated by analyzing the effects of microaerobic conditions on gene expression. PMID- 12571050 TI - Differences between Pseudomonas syringae pv. syringae B728a and Pantoea agglomerans BRT98 in epiphytic and endophytic colonization of leaves. AB - The leaf colonization strategies of two bacterial strains were investigated. The foliar pathogen Pseudomonas syringae pv. syringae strain B728a and the nonpathogen Pantoea agglomerans strain BRT98 were marked with a green fluorescent protein, and surface (epiphytic) and subsurface (endophytic) sites of bean and maize leaves in the laboratory and the field were monitored to see if populations of these strains developed. The populations were monitored using both fluorescence microscopy and counts of culturable cells recovered from nonsterilized and surface-sterilized leaves. The P. agglomerans strain exclusively colonized epiphytic sites on the two plant species. Under favorable conditions, the P. agglomerans strain formed aggregates that often extended over multiple epidermal cells. The P. syringae pv. syringae strain established epiphytic and endophytic populations on asymptomatic leaves of the two plant species in the field, with most of the P. syringae pv. syringae B728a cells remaining in epiphytic sites of the maize leaves and an increasing number occupying endophytic sites of the bean leaves in the 15-day monitoring period. The epiphytic P. syringae pv. syringae B728a populations appeared to originate primarily from multiplication in surface sites rather than from the movement of cells from subsurface to surface sites. The endophytic P. syringae pv. syringae B728a populations appeared to originate primarily from inward movement through the stomata, with higher levels of multiplication occurring in bean than in maize. A rainstorm involving a high raindrop momentum was associated with rapid growth of the P. agglomerans strain on both plant species and with rapid growth of both the epiphytic and endophytic populations of the P. syringae pv. syringae strain on bean but not with growth of the P. syringae pv. syringae strain on maize. These results demonstrate that the two bacterial strains employed distinct colonization strategies and that the epiphytic and endophytic population dynamics of the pathogenic P. syringae pv. syringae strain were dependent on the plant species, whereas those of the nonpathogenic P. agglomerans strain were not. PMID- 12571051 TI - Characterization of a new operon, as-48EFGH, from the as-48 gene cluster involved in immunity to enterocin AS-48. AB - Enterocin AS-48 is a cyclic peptide produced by Enterococcus faecalis S-48 whose genetic determinants have been identified in the conjugative plasmid pMB2. A region of 7.8 kb, carrying the minimum information required for production of and immunity against AS-48, had been previously cloned and sequenced in pAM401 (pAM401-52). In this region, the as-48A structural gene and as-48B, as-48C, as 48C(1), as-48D, and as-48D(1) genes and open reading frame 6 (ORF6) and ORF7 had been identified. The sequence analysis carried out in this work in the BglII B fragment (6.6-kb) from pMB2 cloned downstream from the last ORF identified (ORF7) revealed the existence of two new ORFs, as-48G and as-48H, necessary for full AS 48 expression. Thus, JH2-2 transformants obtained with the pAM401-81 plasmid became producers and resistant at the wild-type level. Tn5 disruption experiments in the last genes, as-48EFGH, were not able to reproduce these expression levels, confirming that expression of these genes is necessary to get the phenotype conferred by the wild-type pMB2 plasmid. The as-48EFGH operon encodes a new ABC transporter that could be involved in producer self-protection. On the basis of the observed similarities, As-48G would be the ATP-binding domain, the deduced amino acid sequences of As-48E and As48-H could be assigned as transmembrane subunits, and As-48F, with an N-terminal transmembrane segment and a coiled-coil domain, strongly resembles the structure of some known ABC transporter accessory proteins whose localization in the cell is discussed. This cluster of genes is expressed by two polycistronic mRNAs, T(2) and T(3), in JH2-2(pAM401-81) in coordinate expression. Our results also suggest that expression of T(3) could be regulated, because in JH2-2(pAM401(EH)) transformants, T(3) was not detected, suggesting that these genes do not by themselves confer immunity, in accordance with the requirement for the as-48D(1) gene for immunity against AS-48. PMID- 12571052 TI - Improvement of posttranslational bottlenecks in the production of penicillin amidase in recombinant Escherichia coli strains. AB - Using periplasmic penicillin amidase (PA) from Escherichia coli ATCC 11105 as a model recombinant protein, we reviewed the posttranslational bottlenecks in its overexpression and undertook attempts to enhance its production in different recombinant E. coli expression hosts. Intracellular proteolytic degradation of the newly synthesized PA precursor and translocation through the plasma membrane were determined to be the main posttranslational processes limiting enzyme production. Rate constants for both intracellular proteolytic breakdown (k(d)) and transport (k(t)) were used as quantitative tools for selection of the appropriate host system and cultivation medium. The production of mature active PA was increased up to 10-fold when the protease-deficient strain E. coli BL21(DE3) was cultivated in medium without a proteinaceous substrate, as confirmed by a decrease in the sum of the constants k(d) and k(t). The original signal sequence of pre-pro-PA was exchanged with the OmpT signal peptide sequence in order to increase translocation efficiency; the effects of this change varied in the different E. coli host strains. Furthermore, we established that simultaneous coexpression of the OmpT pac gene with some proteins of the Sec export machinery of the cell resulted in up to threefold-enhanced PA production. In parallel, we made efforts to increase PA flux via coexpression with the kil gene (killing protein). The primary effects of the kil gene were the release of PA into the extracellular medium and an approximately threefold increase in the total amount of PA produced per liter of bacterial culture. PMID- 12571053 TI - Aminopeptidase C of Aspergillus niger is a novel phenylalanine aminopeptidase. AB - A novel enzyme with a specific phenylalanine aminopeptidase activity (ApsC) from Aspergillus niger (CBS 120.49) has been characterized. The derived amino acid sequence is not similar to any previously characterized aminopeptidase sequence but does share similarity with some mammalian acyl-peptide hydrolase sequences. ApsC was found to be most active towards phenylalanine beta-naphthylamide (F-beta NA) and phenylalanine para-nitroanilide (F-pNA), but it also displayed activity towards other amino acids with aromatic side chains coupled to beta NA; other amino acids with non-aromatic side chains coupled to either pNA or beta NA were not hydrolyzed or were poorly hydrolyzed. ApsC was not able to hydrolyze N acetylalanine-pNA, a substrate for acyl-peptide hydrolases. PMID- 12571054 TI - Application of new primer-enzyme combinations to terminal restriction fragment length polymorphism profiling of bacterial populations in human feces. AB - New primer-enzyme combinations for terminal restriction fragment length polymorphism (T-RFLP) targeting of the 16S rRNA gene were constructed by using the T-RFLP analysis program (designated TAP T-RFLP) located at the Ribosomal Database Project website, and their performance was examined empirically. By using the fluorescently labeled 516f primer (Escherichia coli positions 516 to 532) and 1510r primer (positions 1510 to 1492), the 16S rRNA gene was amplified from human fecal DNA. The resulting amplified product was digested with RsaI plus BfaI or with BslI. When the T-RFLP was carried out with fecal DNAs from eight individuals, eight predominant operational taxonomic units (OTUs) were detected with RsaI and BfaI digestion and 14 predominant OTUs were detected with BslI digestion. The distribution of the OTUs was consistent with the results of the computer simulations with TAP T-RFLP. The T-RFLP analyses of the fecal DNAs from individuals gave characteristic profiles, while the variability of the T-RFLP profiles between duplicate DNA preparations from the same samples were minimal. This new T-RFLP method made it easy to predict what kind of intestinal bacterial group corresponded to each OTU on the basis of the terminal restriction fragment length compared with the conventional T-RFLP and, moreover, made it possible to identify the bacterial species that an OTU represents by cloning and sequencing. PMID- 12571055 TI - Cloning and analysis of a DNA fragment stimulating avermectin production in various Streptomyces avermitilis strains. AB - To isolate a gene for stimulating avermectin production, a genomic library of Streptomyces avermitilis ATCC 31267 was constructed in Streptomyces lividans TK21 as the host strain. An 8.0-kb DNA fragment that significantly stimulated actinorhodin and undecylprodigiosin production was isolated. When wild-type S. avermitilis was transformed with the cloned fragment, avermectin production increased approximately 3.5-fold. The introduction of this fragment into high producer (ATCC 31780) and semi-industrial (L-9) strains also resulted in an increase of avermectin production by more than 2.0- and 1.4-fold, respectively. Subclones were studied to locate the minimal region involved in stimulation of pigmented-antibiotic and avermectin production. An analysis of the nucleotide sequence of the entire DNA fragment identified eight complete and one incomplete open reading frame. All but one of the deduced proteins exhibited strong homology (68 to 84% identity) to the hypothetical proteins of Streptomyces coelicolor A3(2). The orfX gene product showed no significant similarity to any other protein in the databases, and an analysis of its sequence suggested that it was a putative membrane protein. Although the nature of the stimulatory effect is still unclear, the disruption of orfX revealed that this gene was intrinsically involved in the stimulation of avermectin production in S. avermitilis. PMID- 12571056 TI - Molecular characterization of tet(M) genes in Lactobacillus isolates from different types of fermented dry sausage. AB - The likelihood that products prepared from raw meat and milk may act as vehicles for antibiotic-resistant bacteria is currently of great concern in food safety issues. In this study, a collection of 94 tetracycline-resistant (Tc(r)) lactic acid bacteria recovered from nine different fermented dry sausage types were subjected to a polyphasic molecular study with the aim of characterizing the host organisms and the tet genes, conferring tetracycline resistance, that they carry. With the (GTG)(5)-PCR DNA fingerprinting technique, the Tc(r) lactic acid bacterial isolates were identified as Lactobacillus plantarum, L. sakei subsp. carnosus, L. sakei subsp. sakei, L. curvatus, and L. alimentarius and typed to the intraspecies level. For a selection of 24 Tc(r) lactic acid bacterial isolates displaying unique (GTG)(5)-PCR fingerprints, tet genes were determined by means of PCR, and only tet(M) was detected. Restriction enzyme analysis with AccI and ScaI revealed two different tet(M) allele types. This grouping was confirmed by partial sequencing of the tet(M) open reading frame, which indicated that the two allele types displayed high sequence similarities (>99.6%) with tet(M) genes previously reported in Staphylococcus aureus MRSA 101 and in Neisseria meningitidis, respectively. Southern hybridization with plasmid profiles revealed that the isolates contained tet(M)-carrying plasmids. In addition to the tet(M) gene, one isolate also contained an erm(B) gene on a different plasmid from the one encoding the tetracycline resistance. Furthermore, it was also shown by PCR that the tet(M) genes were not located on transposons of the Tn916/Tn1545 family. To our knowledge, this is the first detailed molecular study demonstrating that taxonomically and genotypically diverse Lactobacillus strains from different types of fermented meat products can be a host for plasmid borne tet genes. PMID- 12571057 TI - Identification and characterization of Lactobacillus helveticus PepO2, an endopeptidase with post-proline specificity. AB - A post-proline endopeptidase (PepO2) was detected in cell extracts from a genomic library of Lactobacillus helveticus CNRZ32 by using the synthetic substrate N acetyl-beta-casein-(f203-209)-rho-nitroanilide in a coupled reaction with aminopeptidase N. Isolates with activity for this substrate contained plasmids with visually indistinguishable restriction profiles. Nucleotide sequence analysis revealed a 1,947-bp open reading frame, designated pepO2, encoding a putative 71.4-kDa protein. Analysis of the predicted peptide sequence revealed that L. helveticus PepO2 contained the zinc-dependent metalloprotease motif HEXXH and exhibited levels of amino acid sequence similarity of 72, 61, 59, and 53% to L. helveticus PepO, Lactococcus lactis PepO2, L. lactis PepO, and Lactobacillus rhamnosus PepO, respectively. Northern hybridization results indicated that the transcript containing pepO2 was monocistronic. Despite the high degrees of amino acid similarity to PepO proteins from other lactic acid bacteria, the specificity of the L. helveticus PepO2 for post-proline bonds distinguishes it from other PepO-type endopeptidases characterized to date. The specificity for post-proline bonds also suggests that this enzyme may play a central role in the hydrolysis of casein-derived bitter peptides, such as beta-casein(f193-209). PMID- 12571058 TI - Hydrolysis of casein-derived peptides alpha(S1)-casein(f1-9) and beta-casein(f193 209) by Lactobacillus helveticus peptidase deletion mutants indicates the presence of a previously undetected endopeptidase. AB - Peptides derived from hydrolysis of alpha(S1)-casein(f1-9) [alpha(S1)-CN(f1-9)] and beta-CN(f193-209) with cell extracts of Lactobacillus helveticus CNRZ32 and single-peptidase mutants (Delta pepC, Delta pepE, Delta pepN, Delta pepO, and Delta pepX) were isolated by using reverse-phase high-performance liquid chromatography and were characterized by mass spectrometry. The peptides identified suggest that there was activity of an endopeptidase, distinct from previously identified endopeptidases (PepE and PepO), with specificity for peptide bonds C terminal to Pro residues. Identification of hydrolysis products derived from a carboxyl-blocked form of beta-CN(f193-209) confirmed that the peptides were derived from the activity of an endopeptidase. PMID- 12571060 TI - Enhancement of population size of a biological control agent and efficacy in control of bacterial speck of tomato through salicylate and ammonium sulfate amendments. AB - Sodium salicylate and ammonium sulfate were applied to leaf surfaces along with suspensions of the biological control agents Pseudomonas syringae Cit7(pNAH7), which catabolizes salicylate, and Cit7, which does not catabolize salicylate, to determine whether enhanced biological control of bacterial speck of tomato could be achieved. Foliar amendment with salicylate alone significantly enhanced the population size and the efficacy of Cit7(pNAH7), but not of Cit7, on tomato leaves. Application of ammonium sulfate alone did not result in enhanced population size or biological control efficacy of either Cit7(pNAH7) or Cit7; however, when foliar amendments with both sodium salicylate and ammonium sulfate were applied, a trend toward further increases in population size and biological control efficacy of Cit7(pNAH7) was observed. This study demonstrates the potential of using a selective carbon source to improve the efficacy of a bacterial biological control agent in the control of a bacterial plant disease and supports previous conclusions that the growth of P. syringae in the phyllosphere is primarily carbon limited and secondarily nitrogen limited. PMID- 12571059 TI - Impaired temperature stress response of a Streptococcus thermophilus deoD mutant. AB - An insertional deoD mutant of Streptococcus thermophilus strain SFi39 had a reduced growth rate at 20 degrees C and an enhanced survival capacity to heat shock compared to the wild type, indicating that the deoD product is involved in temperature shock adaptation. We report evidence that ppGpp is implicated in this dual response. PMID- 12571061 TI - Enhanced production of recombinant proteins in Escherichia coli by filamentation suppression. AB - During growth of high-cell-density cultures of Escherichia coli, overproduction of recombinant proteins often results in increased stress response, cell filamentation, and growth cessation. Filamentation of cells consequently lowers final achievable cell concentration and productivity of the target protein. Reported here is a methodology that should prove useful for the enhancement of cell growth and protein productivity by the suppression of cell filamentation. By the coexpression of the E. coli ftsA and ftsZ genes, which encode key proteins in cell division, growth of recombinant strains as well as production of human leptin and human insulin-like growth factor I was improved. Observation of cell morphology revealed that the coexpression of the ftsA and ftsZ genes successfully suppressed filamentation caused by the accumulation of recombinant proteins. PMID- 12571062 TI - High rate of uptake of organic nitrogen compounds by Prochlorococcus cyanobacteria as a key to their dominance in oligotrophic oceanic waters. AB - Direct evidence that marine cyanobacteria take up organic nitrogen compounds in situ at high rates is reported. About 33% of the total bacterioplankton turnover of amino acids, determined with a representative [(35)S]methionine precursor and flow sorting, can be assigned to Prochlorococcus spp. and 3% can be assigned to Synechococcus spp. in the oligotrophic and mesotrophic parts of the Arabian Sea, respectively. This finding may provide a mechanism for Prochlorococcus' competitive dominance over both strictly autotrophic algae and other bacteria in oligotrophic regions sustained by nutrient remineralization via a microbial loop. PMID- 12571063 TI - Studies on the mode of action of reutericyclin. AB - The mode of action of reutericyclin was determined with fluorescent dyes that probed the permeability of the cytoplasmic membrane by large molecules, protons, and potassium. A comparison of reutericyclin activity with those of nisin, nigericin, and valinomycin demonstrated that reutericyclin does not form pores but selectively dissipates the transmembrane proton potential. PMID- 12571064 TI - Cloning and expression analysis of the pcbAB-pcbC beta-lactam genes in the marine fungus Kallichroma tethys. AB - Here we report the identification of the beta-lactam biosynthesis genes pcbAB and pcbC from a cosmid genomic DNA library of the marine fungus Kallichroma tethys. A BLAST homology search showed that they share high sequence identity with the delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV) synthetases and isopenicillin N synthases, respectively, of various fungal and bacterial beta lactam producers, while phylogenetic analysis indicated a close relationship with homologous genes of the cephalosporin-producing pyrenomycete Acremonium chrysogenum. Expression analysis by reverse transcription-PCR suggested that both genes are highly regulated and are expressed in the late growth phase of K. tethys cultures. Complementation of an Aspergillus nidulans strain deficient in ACV synthetase suggested that at least pcbAB is functional, although attempts to isolate active antibiotic from K. tethys were unsuccessful. PMID- 12571065 TI - Genetic differences between two strains of Xylella fastidiosa revealed by suppression subtractive hybridization. AB - Suppression subtractive hybridization was used to rapidly identify 18 gene differences between a citrus variegated chlorosis (CVC) strain and a Pierce's disease of grape (PD) strain of Xylella fastidiosa. The results were validated as being highly representative of actual differences by comparison of the completely sequenced genome of a CVC strain with that of a PD strain. PMID- 12571066 TI - Dibenzyl sulfide metabolism by white rot fungi. AB - Microbial metabolism of organosulfur compounds is of interest in the petroleum industry for in-field viscosity reduction and desulfurization. Here, dibenzyl sulfide (DBS) metabolism in white rot fungi was studied. Trametes trogii UAMH 8156, Trametes hirsuta UAMH 8165, Phanerochaete chrysosporium ATCC 24725, Trametes versicolor IFO 30340 (formerly Coriolus sp.), and Tyromyces palustris IFO 30339 all oxidized DBS to dibenzyl sulfoxide prior to oxidation to dibenzyl sulfone. The cytochrome P-450 inhibitor 1-aminobenzotriazole eliminated dibenzyl sulfoxide oxidation. Laccase activity (0.15 U/ml) was detected in the Trametes cultures, and concentrated culture supernatant and pure laccase catalyzed DBS oxidation to dibenzyl sulfoxide more efficiently in the presence of 2,2' azinobis(3-ethylbenzthiazoline-6-sulfonate) (ABTS) than in its absence. These data suggest that the first oxidation step is catalyzed by extracellular enzymes but that subsequent metabolism is cytochrome P-450 mediated. PMID- 12571067 TI - Chlorine inactivation of spores of Encephalitozoon spp. AB - This report is an extension of a preliminary investigation on the use of chlorine to inactivate spores of Encephalitozoon intestinalis and to investigate the effect of chlorine on two other species, E cuniculi and E. hellem, associated with human infection. The 50% tissue culture infective doses of these three species were also determined. On the basis of the results obtained, it appears that chlorination of water is an effective means of controlling spores of these organisms in the aquatic environment. PMID- 12571068 TI - UV resistance of Bacillus anthracis spores revisited: validation of Bacillus subtilis spores as UV surrogates for spores of B. anthracis Sterne. AB - Recent bioterrorism concerns have prompted renewed efforts towards understanding the biology of bacterial spore resistance to radiation with a special emphasis on the spores of Bacillus anthracis. A review of the literature revealed that B. anthracis Sterne spores may be three to four times more resistant to 254-nm wavelength UV than are spores of commonly used indicator strains of Bacillus subtilis. To test this notion, B. anthracis Sterne spores were purified and their UV inactivation kinetics were determined in parallel with those of the spores of two indicator strains of B. subtilis, strains WN624 and ATCC 6633. When prepared and assayed under identical conditions, the spores of all three strains exhibited essentially identical UV inactivation kinetics. The data indicate that standard UV treatments that are effective against B. subtilis spores are likely also sufficient to inactivate B. anthracis spores and that the spores of standard B. subtilis strains could reliably be used as a biodosimetry model for the UV inactivation of B. anthracis spores. PMID- 12571070 TI - Genomics and homeostasis. AB - The Cannon lecture this year illustrates how knowledge of DNA sequences of complex living organisms is beginning to shape the landscape of physiology in the 21st century. Enormous challenges and opportunities now exist for physiologists to relate the galaxy of genes to normal and pathological functions. The first extensive genomic systems biology map for cardiovascular and renal function was completed last year as well as a new hypothesis-generating tool ("physiological profiling") that enables us to hypothesize relationships between specific genes responsible for the regulation of regulatory pathways. Techniques of chromosomal substitution (consomic and congenic rats) are beginning to confirm statistical results from linkage analysis studies, narrow the regions of genetic interest for positional cloning, and provide genetically well-defined control strains for physiological studies. Patterns of gene expression identified by microarray and mapping of expressed genes to chromosomal sites are adding to the understanding of systems physiology. The previously unimaginable goal of connecting approximately 36,000 genes to the complex functions of mammalian systems is indeed well underway. PMID- 12571069 TI - Recombinant strain of Bacillus thuringiensis producing Cyt1A, Cry11B, and the Bacillus sphaericus binary toxin. AB - A novel recombinant Bacillus thuringiensis subsp. israelensis strain that produces the B. sphaericus binary toxin, Cyt1Aa, and Cry11Ba is described. The toxicity of this strain (50% lethal concentration [LC(50)] = 1.7 ng/ml) against fourth-instar Culex quinquefasciatus was higher than that of B. thuringiensis subsp. israelensis IPS-82 (LC(50) = 7.9 ng/ml) or B. sphaericus 2362 (LC(50) = 12.6 ng/ml). PMID- 12571071 TI - Cardiovascular and renal control in NOS-deficient mouse models. AB - Nitric oxide (NO) plays an essential role in the maintenance of cardiovascular and renal homeostasis. Endogenous NO is produced by three different NO synthase (NOS) isoforms: endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS). To investigate which NOS is responsible for NO production in different tissues, NOS knockout (-/-) mice have been generated for the three isoforms. This review focuses on the regulation of cardiovascular and renal function in relation to blood pressure homeostasis in the different NOS-/- mice. Although regulation of vascular tone and cardiac function in eNOS-/- has been extensively studied, far less is known about renal function in these mice. eNOS-/- mice are hypertensive, but the mechanism responsible for their high blood pressure is still not clear. Less is known about cardiovascular and renal control in nNOS-/- mice, probably because their blood pressure is normal. Recent data suggest that nNOS plays important roles in cardiac function, renal homeostasis, and regulation of vascular tone under certain conditions, but these are only now beginning to be studied. Inasmuch as iNOS is absent from the cardiovascular system under physiological conditions, it may become important to blood pressure regulation only during pathological conditions related to inflammatory processes. However, iNOS is constitutively expressed in the kidney, where its function is largely unknown. Overall, the study of NOS knockout mice has been very useful and produced many answers, but it has also raised new questions. The appearance of compensatory mechanisms suggests the importance of the different isoforms to specific processes, but it also complicates interpretation of the data. In addition, deletion of a single gene may have physiologically significant effects in addition to those being studied. Thus the presence or absence of a specific phenotype may not reflect the most important physiological function of the absent gene. PMID- 12571072 TI - Cardiovascular effects of leptin and orexins. AB - Leptin, the product of the ob gene, is a satiety factor secreted mainly in adipose tissue and is part of a signaling mechanism regulating the content of body fat. It acts on leptin receptors, most of which are located in the hypothalamus, a region of the brain known to control body homeostasis. The fastest and strongest hypothalamic response to leptin in ob/ob mice occurs in the paraventricular nucleus, which is involved in neuroendocrine and autonomic functions. On the other hand, orexins (orexin-A and -B) or hypocretins (hypocretin-1 and -2) were recently discovered in the hypothalamus, in which a number of neuropeptides are known to stimulate or suppress food intake. These substances are considered important for the regulation of appetite and energy homeostasis. Orexins were initially thought to function in the hypothalamic regulation of feeding behavior, but orexin-containing fibers and their receptors are also distributed in parts of the brain closely associated with the regulation of cardiovascular and autonomic functions. Functional studies have shown that these peptides are involved in cardiovascular and sympathetic regulation. The objective of this article is to summarize evidence on the effects of leptin and orexins on cardiovascular function in vivo and in vitro and to discuss the pathophysiological relevance of these peptides and possible interactions. PMID- 12571073 TI - Regulating food intake. PMID- 12571074 TI - Increased albumin plasma efflux contributes to hypoalbuminemia only during early phase of sepsis in rats. AB - The mechanisms leading to hypoalbuminemia in sepsis were explored by measuring plasma volume, albumin distribution, plasma albumin transcapillary escape rate (TER), and efflux (TER x albumin intravascular pool). These parameters were quantified in infected rats, injected intravenously with live Escherichia coli, and pair-fed and well-fed rats using an injection of (35)S-albumin and measuring plasma and whole body albumin concentrations. Animals were studied on days 1, 6, and 10 after infection. In pair-fed rats, neither albumin distribution nor exchange rate between the intra- and extravascular compartments was modified. The increase of plasma volume after infection partly explained hypoalbuminemia. Infection resulted in a reduction of the total albumin pool of the body all along the experimental period, indicating a net loss of the protein. Albumin TER (%/day) was significantly increased 1 and 6 days after infection, but the absolute efflux was increased only on day 1. Normal values were observed on day 10. Therefore, an accelerated plasma efflux contributes to hypoalbuminemia only during the early period of sepsis. During this phase, the protein was retained in the extravascular space where it was probably catabolized. Later on, other factors are probably involved. PMID- 12571075 TI - Exercise elicits phase shifts and acute alterations of melatonin that vary with circadian phase. AB - To examine the immediate phase-shifting effects of high-intensity exercise of a practical duration (1 h) on human circadian phase, five groups of healthy men 20 30 yr of age participated in studies involving no exercise or exposure to morning, afternoon, evening, or nocturnal exercise. Except during scheduled sleep/dark and exercise periods, subjects remained under modified constant routine conditions allowing a sleep period and including constant posture, knowledge of clock time, and exposure to dim light intensities averaging (+/-SD) 42 +/- 19 lx. The nocturnal onset of plasma melatonin secretion was used as a marker of circadian phase. A phase response curve was used to summarize the phase shifting effects of exercise as a function of the timing of exercise. A significant effect of time of day on circadian phase shifts was observed (P < 0.004). Over the interval from the melatonin onset before exercise to the first onset after exercise, circadian phase was significantly advanced in the evening exercise group by 30 +/- 15 min (SE) compared with the phase delays observed in the no-exercise group (-25 +/- 14 min, P < 0.05). Phase shifts in response to evening exercise exposure were attenuated on the second day after exercise exposure and no longer significantly different from phase shifts observed in the absence of exercise. Unanticipated transient elevations of melatonin levels were observed in response to nocturnal exercise and in some evening exercise subjects. Taken together with the results from previous studies in humans and diurnal rodents, the current results suggest that 1) a longer duration of exercise exposure and/or repeated daily exposure to exercise may be necessary for reliable phase-shifting of the human circadian system and that 2) early evening exercise of high intensity may induce phase advances relevant for nonphotic entrainment of the human circadian system. PMID- 12571076 TI - Role of leptin and leptin receptor in inflammation. PMID- 12571077 TI - Hyperleptinemia and reduced TNF-alpha secretion cause resistance of db/db mice to endotoxin. AB - Leptin deficiency in ob/ob mice increases susceptibility to endotoxic shock, whereas leptin pretreatment protects them against LPS-induced lethality. Lack of the long-form leptin receptor (Ob-Rb) in db/db mice causes resistance. We tested the effects of LPS in C57BL/6J db(3J)/db(3J) (BL/3J) mice, which express only the circulating leptin receptors, compared with C57BL/6J db/db (BL/6J) mice, which express all short-form and circulating isoforms of the leptin receptor. Intraperitoneal injections of LPS significantly decreased rectal temperature and increased leptin, corticosterone, and free TNF-alpha in fed and fasted BL/3J and BL/6J mice. TNF-alpha was increased three- and fourfold in BL/3J and BL/6J, respectively. LPS (100 microg) caused 50% mortality of fasted BL/6J mice but caused no mortality in fasted BL/3J mice. Pretreatment of fasted BL/3J mice with 30 microg leptin prevented the drop in rectal temperature, blunted the increase in corticosterone, but had no effect on TNF-alpha induced by 100 microg LPS. Taken together, these data provide evidence that fasted BL/3J mice are more resistant than BL/6J mice to LPS toxicity, presumably due to the absence of leptin receptors in BL/3J mice. This resistance may be due to high levels of free leptin cross-reacting with other cytokine receptors. PMID- 12571078 TI - What is the role of serotonin during hemorrhage in conscious animals? PMID- 12571079 TI - Reloading of atrophied rat soleus muscle induces tenascin-C expression around damaged muscle fibers. AB - The hypothesis was tested that mechanical loading, induced by hindlimb suspension and subsequent reloading, affects expression of the basement membrane components tenascin-C and fibronectin in the belly portion of rat soleus muscle. One day of reloading, but not the previous 14 days of hindlimb suspension, led to ectopic accumulation of tenascin-C and an increase of fibronectin in the endomysium of a proportion (8 and 15%) of muscle fibers. Large increases of tenascin-C (40-fold) and fibronectin (7-fold) mRNA within 1 day of reloading indicates the involvement of pretranslational mechanisms in tenascin-C and fibronectin accumulation. The endomysial accumulation of tenascin-C was maintained up to 14 days of reloading and was strongly associated with centrally nucleated fibers. The observations demonstrate that an unaccustomed increase of rat soleus muscle loading causes modification of the basement membrane of damaged muscle fibers through ectopic endomysial expression of tenascin-C. PMID- 12571080 TI - A new two-breath technique for extracting the cerebrovascular response to arterial carbon dioxide. AB - Cerebrovascular autoregulation is evaluated from spontaneous fluctuations in mean flow velocity (MFV) by transcranial Doppler ultrasound of the middle cerebral artery (MCA) with respect to changes in arterial blood pressure (BP(MCA)), but the effects of spontaneous fluctuations in arterial Pco(2) on MFV have been largely ignored. Autoregressive moving average analysis (ARMA), a closed-loop system identification technique, was applied to data from nine healthy subjects during spontaneous breathing, during inspiration of 10% CO(2) for two breaths once per minute for 4 min, and during sustained breathing of 7% CO(2). Cerebrovascular resistance index (CVRi) was calculated (CVRi = BP(MCA)/MFV). Reliable estimates of gain for BP(MCA) --> MFV were obtained for spontaneous breathing and the two-breath method. In contrast, reliable gain estimates for Pco(2) --> MFV or Pco(2) --> CVRi were achieved only under the two-breath method. Pco(2) --> MFV gain was smaller with the two-breath method than during sustained 7% CO(2) (P < 0.05). BP(MCA) was elevated by 7% CO(2) but not by the two-breath method. The closed-loop model provides insight into interactions between BP(MCA) and Pco(2) on cerebrovascular control, but reliable solutions for Pco(2) effects with ARMA analysis require perturbation by the two-breath method. PMID- 12571081 TI - Near-term suppression of fever: inhibited synthesis or accelerated catabolism of prostaglandin E2? PMID- 12571082 TI - Conjugated linoleic acid inhibits cell proliferation and ErbB3 signaling in HT-29 human colon cell line. AB - Conjugated linoleic acid (CLA) has chemoprotective properties in experimental cancer models, and in vitro studies have shown that CLA inhibits HT-29 colon cancer cell growth. ErbB2 and ErbB3 have been implicated in the development of colon cancer, and both proteins are expressed at high levels in the HT-29 cell line. Activation of ErbB2/ErbB3 heterodimers is regulated by the ErbB3 ligand heregulin. To examine CLA regulation of HT-29 cell proliferation and apoptosis and the influence of CLA on the ErbB3 signaling pathway, HT-29 cells were cultured in the presence of CLA and/or heregulin. CLA inhibited DNA synthesis and induced apoptosis of HT-29 cells. Although the addition of heregulin-alpha led to an increase in cell number, it was not able to counteract the negative growth regulatory effect of CLA. Immunoprecipitation/Western blot studies revealed that CLA inhibited heregulin-alpha-stimulated phosphorylation of ErbB2 and ErbB3, recruitment of the p85 subunit of phosphoinositide 3-kinase (PI3-kinase) to the ErbB3 receptor, ErbB3-associated PI3-kinase activities, and phosphorylation of Akt. CLA decreased ErbB2 and ErbB3 mRNA and protein levels in a dose-dependent manner. In conclusion, we demonstrate that CLA inhibits cell proliferation and stimulates apoptosis in HT-29 cells and that this may be mediated by its ability to downregulate ErbB3 signaling and the PI3-kinase/Akt pathway. PMID- 12571083 TI - Hemodynamic and proinflammatory actions of endothelin-1 in guinea pig small intestine submucosal microcirculation. AB - The hemodynamic and proinflammatory effects of endothelin-1 (ET-1) in proximal (1st/2nd order) and terminal (3rd/4th order) arterioles and venules were examined in small intestine submucosa of anesthetized guinea pigs. Vessel diameter (D), red blood cell velocity, and blood flow (Q) were determined in eight proximal and eight terminal microvessels before and at 20 min of ET-1 suffusion (10(-10), 10( 9), and 10(-8) M) and then with endothelin-A (ET(A))-receptor blockade with BQ 123 (10(-5) M). This protocol was repeated with platelet-activating factor (PAF) inhibition (WEB-2086, 1.0 mg/kg iv; n = 16). The ET-1-mediated microvascular responses were also examined with endothelin-B (ET(B))-receptor blockade using BQ 788 (10(-5) M; n = 11) alone or with ET(A+B)-receptor blockade with BQ-123 + BQ 788 (n = 10). Microvascular permeability was assessed by FITC-albumin (25 mg/kg iv) extravasation in seven series: 1) buffered modified Krebs solution suffusion (n = 6), 2) histamine suffusion (HIS; 10(-3) M, n = 5), 3) ET-1 suffusion (10(-8) M, n = 5), 4) BQ-123 (10(-5) M) plus ET-1 suffusion (n = 5), 5) PAF inhibition before ET-1 suffusion (n = 5), 6) histamine-1 (H1)-receptor blockade (diphenhydramine, 20 mg/kg iv) before ET-1 suffusion (n = 5), and 7) ET(B) receptor blockade before (BQ-788 10(-5) M; n = 3) or with ET-1 suffusion (n = 3). D and Q decreased at 10(-8) M ET-1 and returned to control values with BQ-123 and BQ-123+BQ788 but not with BQ-788 in proximal microvessels. D did not change in terminal microvessels with ET-1 (10(-8) M) but decreased with BQ-788 and increased with BQ-123. PAF inhibition did not affect the D and Q responses of proximal microvessels to ET-1 but prevented the fall in Q in terminal microvessels with ET-1. ET-1 increased vascular permeability to approximately 1/3 of that with HIS; this response was prevented with BQ-123 and WEB-2086 but not with H1-receptor blockade. This is the first evidence that submucosal terminal microvessel flow is reduced with ET-1 independent of vessel diameter changes and that this response is associated with increased microvascular permeability mediated via ET(A)-receptor stimulation and PAF activation. PMID- 12571084 TI - Modulation of intestinal protein synthesis and protease mRNA by luminal and systemic nutrients. AB - Route of nutrient supply is important in regulation of intestinal protein metabolism, because total parenteral nutrition, compared with enteral feeding, leads to profound atrophy. Participation of the fractional rate of protein synthesis (Ks), their degradation in regulation of gut protein balance, and their possible modulation by specific nutrients are the focus of our work. We developed an in situ experimental system that allows controlled exposure of intestinal mucosa to nutrients systemically, luminally, or both. We examined the effects of systemic glucose and amino acid (AA) infusion in overnight-fasted piglets. Jejunal segments within each piglet were simultaneously, luminally perfused with solutions containing various AAs or glucose. Intravenous infusion of glucose increased mucosal Ks by 16% (P < 0.05), whereas intravenous infusion of AA had no effect on Ks. Systemic glucose infusion had no effect on mRNA levels for components of the ubiquitin-proteasome proteolytic pathway. However, levels of these mRNA were reduced by intravenous or luminal AA supply. This effect was greatest (-50%) when highest tissue concentrations of AAs were achieved by the simultaneous infusion of AA by both routes (P < 0.05). Our findings suggest that not only is the modulation of protein balance in the intestine in response to nutrients in part attributable to anabolic stimulation of protein synthesis initiated by the systemic appearance of glucose, but a fall in protein degradation is also a likely contributor. AAs appear to be a key factor required to reduce expression of genes connected with proteolysis. PMID- 12571085 TI - High-level activation by a duodenum-specific enhancer requires functional GATA binding sites. AB - The purine metabolic gene adenosine deaminase (ADA) is expressed at high levels in a well-defined spatiotemporal pattern in the villous epithelium of proximal small intestine. A duodenum-specific enhancer module responsible for this expression pattern has been identified in the second intron of the human ADA gene. It has previously been shown that binding of the factor PDX-1 is essential for function of this enhancer. The studies presented here examine the proposed roles of GATA factors in the enhancer. Site-directed mutagenesis of the enhancer's GATA binding sites crippled enhancer function in 10 lines of transgenic mice, with 9 of the lines demonstrating <1% of normal activity. Detailed studies along the longitudinal axis of mouse small intestine indicate that GATA-4 and GATA-5 mRNA levels display a reciprocal pattern, with low levels of GATA-6 throughout. Interestingly, gel shift studies with duodenal nuclear extracts showed binding only by GATA-4. PMID- 12571086 TI - Promoting healthy weight--the new environmental frontier. PMID- 12571087 TI - Holding fast: the experience of collaboration in a competitive environment. AB - Collaboration is one of the cornerstones of health promotion, with the literature indicating a range of circumstances under which it can either succeed or be undermined. In New Zealand in the 1990s, a market structure for health made collaboration of all kinds exceptionally difficult. This paper traces the efforts of a group of nutrition agencies (Agencies for Nutrition Action) to defy the popular wisdom and persist with collaborative efforts. The agencies were unsuccessful in their attempts to develop joint campaigns, but were very successful in advocacy and intersectoral action that did not threaten the position of individual agencies in the competitive environment. It is possible that the collaboration could have been more effective if agencies had been willing to surrender some autonomy and commit themselves to supporting a more independent new organization. However, this would have compromised not only their individual integrity but also their commitment to a relationship of equals. In 'holding fast' to a belief in health promotion, the ANA resisted being coopted by a now discredited market system, and emerged with its integrity and that of its participating agencies intact. ANA is now well positioned to work within an emerging policy environment that is more supportive of health promotion. PMID- 12571088 TI - Health promotion partnerships in Israel: motives, enhancing and inhibiting factors, and modes of structure. AB - Multisectoral cooperation is an important strategy in working for health promotion. Fifty-two health professionals completed a questionnaire measuring factors motivating, enhancing and inhibiting partnerships. The respondents also reported the type or structure of the partnerships. The results indicated that partnerships were formed primarily in order to promote the project; however, previous positive experience with partnerships was also very important as a motivating force for joining partnerships. The three most important facilitating factors were related to project management: effective leadership, aims of the project, and sharing a vision and goals. The two most frequent items mentioned as very important barriers to partnerships were related to dysfunction of the steering committee. Two types or structures of partnerships were identified. The first was fragmental, where partnerships existed only at specific stages of the project; most respondents reported working within this structure. The second type was continuous, where partnerships existed through all the project's stages. Using multiple regression analysis we found that health staff (mainly nurses) worked more frequently in fragmental partnership structures than did health promotion and welfare workers. In addition, the more experienced the respondents, the less they worked in fragmental partnership structures. These results highlight the importance of acquiring skills for working in partnerships and indicate a need for guidelines to be agreed by the partners at the beginning of the working process. PMID- 12571089 TI - Community participation for perinatal health in Istanbul. AB - This study aimed to evaluate the success of a project in achieving community participation in efforts to improve perinatal health. A 10-step structured process was used to work with a community in Istanbul, Turkey. To evaluate the success of the project in achieving community participation, five key indicators were selected: (i) participation of the community group in decision making; (ii) gains in knowledge and skills of the community group; (iii) continuity of the community group; (iv) continuation of the health program by the community group; and (v) initiation of new support and advocacy activities. From the beginning, community members participated in all decisions regarding the group activities. In the early months, project staff had more of a guiding role, but, over time, the community members became active decision-makers. Over the course of the project they learned how to identify community health problems, and to design, implement and evaluate interventions to address those problems. Four years later, meetings and activities of the group are continuing. The antenatal education course developed by the group continues to be offered at a local community centre. Community members are now completely responsible for promoting the course, communication with participants, planning the courses, making preparations for the sessions, as well as teaching non-technical topics. Course participants have developed ongoing support networks and have begun to advocate for better perinatal health services in the community. Working with a community is an intensive, time-consuming process. The results of this project indicate that the benefits, both in terms of health outcomes and in terms of increases in community capacity, can be well worth the effort. PMID- 12571090 TI - Empowering social action through narratives of identity and culture. AB - Concern at widening health and wealth inequities between communities accompanying processes of globalization in recent years are reflected in contemporary definitions of health promotion, premised on the stratagem of individuals and communities increasing control over factors that determine health, thereby improving their health status. Such community empowerment practice is commonly accepted within the health promotion literature as encompassing intrapersonal, interpersonal and socio-political elements. Less articulated and understood, however, are the processes whereby the identities and cultures of marginalized communities intersect with and reverberate through these levels of action. The potential of identity and culture as important individual and community resources within social action takes on further significance within global-ized contexts, which simultaneously expose marginalized communities to dominant cultural power relations while affording members new avenues for cultural expression. In this paper we highlight culture and identity as important aspects of the empowerment process, drawing on the experiences of migrant Tongan and Samoan women throughout a social action process in Aotearoa/New Zealand. In particular, narratives of identity and culture within storytelling as an empowerment practice are explicated, as is the articulation of identity and culture within more structurally orientated power relations throughout subsequent activities related to policy advocacy. PMID- 12571091 TI - Reaching hard-to-reach, high-risk populations: piloting a health promotion and diabetes disease prevention programme on an urban marae in New Zealand. AB - Maori and other indigenous peoples experience a high prevalence of type 2 diabetes. A pivotal question is how primary and secondary preventative initiatives might be more effectively targeted to embrace those who are at highest risk of developing diabetes and its complications. This paper proposes that, in the case of Maori, as a high-risk population, conventional approaches are insufficient, and that increased consideration needs to be given to how the settings in which health education and services are offered may influence diabetes prevention and earlier diagnosis. Traditionally, the hub of Maori culture and everyday life is the marae, a place where Maori identity, values and cultural practices are affirmed within an over-arching spiritual dimension. We have investigated the potential utility of an urban marae and its member network as a setting for a lifestyle programme focused around diabetes prevention. The research included a cross sectional survey of behavioural and metabolic risk factors for type 2 diabetes and qualitative data collection as part of a formative and process evaluation of a lifestyle programme established at the marae and connected venues. The programme attracted 436 participants. The majority knew little about diabetes, had low levels of vigorous activity and high intakes of fatty foods. A family history of diabetes was present in >40% of participants. Undiagnosed diabetes, high blood pressure, hypercholesterolaemia, obesity, smoking and self-reported excessive alcohol consumption were common. The advent of diabetes education, a healthy lifestyle support programme, and exercise sessions at the marae and connected venues served as the impetus for the marae community to take over the running of their own health promotion programme, including a declaration of their marae as a 'smoke-free' venue. It is proposed that marae can be useful settings for lifestyle programmes aimed at controlling the diabetes and obesity epidemic in New Zealand. PMID- 12571092 TI - A case study of sodium reduction in breakfast cereals and the impact of the Pick the Tick food information program in Australia. AB - In 1997, one of Australia's largest food companies undertook a program of salt reduction in 12 breakfast cereals. The National Heart Foundation's Pick the Tick program criterion (<400 mg sodium per 100 g) was used as a target value where possible. Twelve products were reformulated, with reductions ranging from 85 to 479 mg sodium per 100 g and an average reduction of 40% (12-88%). As a result, 235 tonnes of salt were removed annually from the Australian food supply and five more products were able to carry the tick logo. The impact of the Pick the Tick program in changing the food supply extends beyond those products that are part of the food approval program. PMID- 12571093 TI - The association between health risk status and health care costs among the membership of an Australian health plan. AB - Health promotion in Australia has developed into an accepted strategy for solving public health problems and promoting the health of its citizens. However, there are few evidence-based research studies in Australia that measure health risk status or track health changes over time with defined cost outcome measures. Those individuals with more high-risk lifestyle behaviors have been associated with higher costs compared with those with low-risk behaviors. Although intuitively it was believed that the health promotion programs had a positive impact on health behaviors and consequently on health care costs, the relationship between health risk status and health care costs had yet to be tested in the Australian population. Consequently, a verification study was initiated by the Australian Health Management Group (AHMG) to confirm that those relationships between health risks and medical costs that had been published would also hold in the Australian population using Australian private health care costs as the outcome measure. Eight health risks were defined using a Health Risk Appraisal (HRA) to determine the health risk status of participants. Consistent with previous studies, low-risk participants were associated with the lowest health care costs (377 Australian dollars) compared with medium- (484 Australian dollars) or high-risk (661 Australian dollars) participants and non-participants (438 Australian dollars). If the health care costs of those at low risk were considered as the baseline costs, excess health care costs associated with excess health risks in this population were calculated at 13.5% of total expenditures. Health risk reduction and low-risk maintenance can provide important strategies for improving/maintaining the health and well-being of the membership and for potential savings in health care costs. PMID- 12571094 TI - Using patient-driven computers to provide cost-effective prevention in primary care: a conceptual framework. AB - This paper initially presents a rationale for the cost-effectiveness of using patient-driven computers in primary care services. It specifically defines the concepts of prevention and primary care, prior to outlining the advantages of promoting the implementation of prevention practices in primary care. It argues that greater use of computer technology represents one means of cost-effectively optimizing the integration of prevention into routine primary care, and identifies an apparent disjuncture between the potential of computers and the limited success with which attempts to integrate them into routine primary care services have been met, as evidenced in the published international literature. Among several possible explanations for this disjuncture, such as a possible lack of precision with which computers identify at-risk patients, perceived high costs associated with computers and physicians' concerns about the inflexibility and the more impersonal nature of computer interactions, is the apparent failure of researchers to utilize well designed and empirically tested models in the planning, implementation and evaluation of computerized care. An outline for such an approach, utilizing the Precede-Proceed model of health promotion planning and the Diffusion of Innovations theory, is presented. PMID- 12571096 TI - Irx4-mediated regulation of Slit1 expression contributes to the definition of early axonal paths inside the retina. AB - Although multiple axon guidance cues have been discovered in recent years, little is known about the mechanism by which the spatiotemporal expression patterns of the axon guidance cues are regulated in vertebrates. We report that a homeobox gene Irx4 is expressed in a pattern similar to that of Slit1 in the chicken retina. Overexpression of Irx4 led to specific downregulation of Slit1 expression, whereas inhibition of Irx4 activity by a dominant negative mutant led to induction of Slit1 expression, indicating that Irx4 is a crucial regulator of Slit1 expression in the retina. In addition, by examining axonal behavior in the retinas with overexpression of Irx4 and using several in vivo assays to test the effect of Slit1, we found that Slit1 acts positively to guide the retinal axons inside the optic fiber layer (OFL). We further show that the regulation of Slit1 expression by Irx4 is important for providing intermediate targets for retinal axons during their growth within the retina. PMID- 12571097 TI - Root or crown: a developmental choice orchestrated by the differential regulation of the epithelial stem cell niche in the tooth of two rodent species. AB - The rodent incisor grows continuously throughout its lifetime. The epithelial stem cell niche is located at the apical end of the tooth and its progeny gives rise to the ameloblasts that form the hard enamel. Previously, mesenchymal FGF10 was shown to support the niche, in conjunction with epithelial Notch signaling. Here we show that in a different continuously growing tooth type, the molar of the sibling vole, a similar regulatory system is in place. Moreover, the identical expression pattern of Bmp4 compared to Fgf10 suggests that BMP4 could also be involved in the regulation of the epithelial stem cell niche. Notch and FGF10 signaling is mainly absent in the mouse molar, which stops growing and develops roots. The regulation of the epithelial stem cell niche seems to be flexible allowing for the existence of different tooth types, such as continuously growing teeth, and high and low crowned molars. PMID- 12571098 TI - The parapineal mediates left-right asymmetry in the zebrafish diencephalon. AB - The dorsal diencephalon (or epithalamus) of larval zebrafish displays distinct left-right asymmetries. The pineal complex consists of the pineal organ anlage and an unpaired, left-sided accessory organ - the parapineal. The neighboring brain nuclei, the left and right dorsal habenulae, show consistent differences in their size, density of neuropil and gene expression. Mutational analyses demonstrate a correlation between the left-right position of the parapineal and the laterality of the habenular nuclei. We show that selective ablation of the parapineal organ results in the loss of habenular asymmetry. The left-sided parapineal therefore influences the left-right identity of adjacent brain nuclei, indicating that laterality of the dorsal diencephalon arises in a step-wise fashion. PMID- 12571099 TI - Targeted deletion of a branchial arch-specific enhancer reveals a role of dHAND in craniofacial development. AB - The basic helix-loop-helix transcription factor dHAND is expressed in the mesenchyme of branchial arches and the developing heart. Mice homozygous for a dHAND (Hand2) null mutation die early in embryogenesis from cardiac abnormalities, precluding analysis of the potential role of dHAND in branchial arch development. Two independent enhancers control expression of dHAND in the heart and branchial arches. Endothelin-1 (ET-1) signaling regulates the branchial arch enhancer and is required for dHAND expression in the branchial arches. To determine the potential role of dHAND in branchial arch development and to assess the role of the ET-1-dependent enhancer in dHAND regulation in vivo, we deleted this enhancer by homologous recombination. Mice lacking the dHAND branchial arch enhancer died perinatally and exhibited a spectrum of craniofacial defects that included cleft palate, mandibular hypoplasia and cartilage malformations. Expression of dHAND was abolished in the ventolateral regions of the first and second branchial arches in these mutant mice, but expression was retained in a ventral domain where the related transcription factor eHAND is expressed. We conclude that dHAND plays an essential role in patterning and development of skeletal elements derived from the first and second branchial arches and that there are heterogeneous populations of cells in the branchial arches that rely on different cis-regulatory elements for activation of dHAND transcription. PMID- 12571100 TI - Structural-proliferative units and organ growth: effects of insulin-like growth factor 2 on the growth of colon and skin. AB - Many epithelial renewal tissues in vertebrates are organised into structural proliferative units. We have examined the effect of IGF2 dose on the structure of structural-proliferative units in skin and colon. The mouse strains used were the Igf2 knockout, wild type and K:Igf2, a transgenic in which Igf2 is overexpressed under control of a keratin promoter. For both skin and colon, the histological organisation of structural-proliferative units was unaltered with increasing IGF2 dose, although there was a higher fraction of dividing cells in the proliferative compartment. In the colon an increase in IGF2 dose increases the overall area of the epithelium. This is due to an increase in the number of crypts with no change of cell size or of crypt area. Growth stimulation appears to be due to a reduction in the duration of crypt fission. The conclusion is that the IGF2 pathway can stimulate the multiplication of colonic crypts independently of stimulating increased cell proliferation. The results for the skin are consistent with this. An increase of IGF2 dose increases the proportion of dividing cells in the basal layer, the thickness of the epidermis and the total area of the epidermis. By comparison with Drosophila, these results show no effects on cell size, but do show the possibility of inducing disproportionate growth. These differences may represent properties of the SPU organisation that is characteristic of vertebrate tissues. PMID- 12571101 TI - Cyclic GMP-dependent protein kinase EGL-4 controls body size and lifespan in C elegans. AB - We designed an automatic system to measure body length, diameters and volume of a C. elegans worm. By using this system, mutants with an increased body volume exceeding 50% were isolated. Four of them are grossly normal in morphology and development, grow longer to be almost twice as big, and have weak egg-laying defects and extended lifespan. All the four mutants have a mutation in the egl-4 gene. We show that the egl-4 gene encodes cGMP-dependent protein kinases. egl-4 promoter::gfp fusion genes are mainly expressed in head neurons, hypodermis, intestine and body wall muscles. Procedures to analyze morphology and volume of major organs were developed. The results indicate that volumes of intestine, hypodermis and muscle and cell volumes in intestine and muscle are increased in the egl-4 mutants, whereas cell numbers are not. Experiments on genetic interaction suggest that the cGMP-EGL-4 signaling pathway represses body size and lifespan through DBL-1/TGF-beta and insulin pathways, respectively. PMID- 12571102 TI - FGF signaling through FGFR1 is required for olfactory bulb morphogenesis. AB - During development, the embryonic telencephalon is patterned into different areas that give rise to distinct adult brain structures. Several secreted signaling molecules are expressed at putative signaling centers in the early telencephalon. In particular, Fgf8 is expressed at the anterior end of the telencephalon and is hypothesized to pattern it along the anteroposterior (AP) axis. Using a CRE/loxP genetic approach to disrupt genes in the telencephalon, we address the role of FGF signaling directly in vivo by abolishing expression of the FGF receptor Fgfr1. In the Fgfr1-deficient telencephalon, AP patterning is largely normal. However, morphological defects are observed at the anterior end of the telencephalon. Most notably, the olfactory bulbs do not form normally. Examination of the proliferation state of anterior telencephalic cells supports a model for olfactory bulb formation in which an FGF-dependent decrease in proliferation is required for initial bulb evagination. Together the results demonstrate an essential role for Fgfr1 in patterning and morphogenesis of the telencephalon. PMID- 12571103 TI - Tlx and Pax6 co-operate genetically to establish the pallio-subpallial boundary in the embryonic mouse telencephalon. AB - We have examined the role of Tlx, an orphan nuclear receptor, in dorsal-ventral patterning of the mouse telencephalon. Tlx is expressed broadly in the ventricular zone, with the exception of the dorsomedial and ventromedial regions. The expression spans the pallio-subpallial boundary, which separates the dorsal (i.e. pallium) and ventral (i.e. subpallium) telencephalon. Despite being expressed on both sides of the pallio-subpallial boundary, Tlx homozygous mutants display alterations in the development of this boundary. These alterations include a dorsal shift in the expression limits of certain genes that abut at the pallio-subpallial boundary as well as the abnormal formation of the radial glial palisade that normally marks this boundary. The Tlx mutant phenotype is similar to, but less severe than, that seen in Small eye (i.e. Pax6) mutants. Interestingly, removal of one allele of Pax6 on the homozygous Tlx mutant background significantly worsens the phenotype. Thus Tlx and Pax6 cooperate genetically to regulate the establishment of the pallio-subpallial boundary. The patterning defects in the Tlx mutant telencephalon result in a loss of region specific gene expression in the ventral-most pallial region. This correlates well with the malformation of the lateral and basolateral amygdala in Tlx mutants, both of which have been suggested to derive from ventral portions of the pallium. PMID- 12571104 TI - Molecular analysis of axon repulsion by the notochord. AB - During development of the amniote peripheral nervous system, the initial trajectory of primary sensory axons is determined largely by the action of axon repellents. We have shown previously that tissues flanking dorsal root ganglia, the notochord lying medially and the dermamyotomes lying laterally, are sources of secreted molecules that prevent axons from entering inappropriate territories. Although there is evidence suggesting that SEMA3A contributes to the repellent activity of the dermamyotome, the nature of the activity secreted by the notochord remains undetermined. We have employed an expression cloning strategy to search for axon repellents secreted by the notochord, and have identified SEMA3A as a candidate repellent. Moreover, using a spectrum of different axon populations to assay the notochord activity, together with neuropilin/Fc receptor reagents to block semaphorin activity in collagen gel assays, we show that SEMA3A probably contributes to notochord-mediated repulsion. Sympathetic axons that normally avoid the midline in vivo are also repelled, in part, by a semaphorin based notochord activity. Although our results implicate semaphorin signalling in mediating repulsion by the notochord, repulsion of early dorsal root ganglion axons is only partially blocked when using neuropilin/Fc reagents. Moreover, retinal axons, which are insensitive to SEMA3A, are also repelled by the notochord. We conclude that multiple factors act in concert to guide axons in this system, and that further notochord repellents remain to be identified. PMID- 12571105 TI - Sox5 and Sox6 are required for notochord extracellular matrix sheath formation, notochord cell survival and development of the nucleus pulposus of intervertebral discs. AB - The notochord has major roles in vertebral column formation: indirectly by inducing sclerotome cell differentiation; and directly by forming the nucleus pulposus of intervertebral discs. Sox5 and Sox6 encode Sry-related HMG box transcription factors that act redundantly to promote chondroblast differentiation in all cartilages of the mouse embryo. We show that Sox5 and Sox6 are expressed in the notochord cell lineage and required for notochord late development. In Sox5(-/-)/Sox6(-/-) embryos, the notochord formed a typical rod like structure. It fulfilled its inductive functions, as indicated by expression of sonic hedgehog and sclerotome specification. However, the notochord failed to become surrounded with an extracellular matrix sheath. This phenotype was associated with a downregulation of extracellular matrix genes, including the genes for collagen 2, aggrecan and perlecan in both notochord cells and surrounding chondrocytic cells of presumptive inner annuli and vertebral bodies. The mutant notochord then underwent an aberrant, fatal dismantling after sclerotome cell migration. Its cells became removed first from intervertebral spaces and then from vertebral bodies, and it progressively underwent apoptosis. Meanwhile, the development of inner annuli and vertebral bodies was dramatically impaired. Consequently, the vertebral column of Sox5(-/-)/Sox6(-/-) fetuses consisted of a very deficient cartilage and was devoid of nuclei pulposi. In Sox5(-/-)/Sox6(+/-) and more severely in Sox5(+/-)/Sox6(-/-) embryos, the notochord sheath was thinner, but cells survived. By birth, nuclei pulposi were rudimentary, and its cells poorly swelled and still expressing sonic hedgehog. Hence, Sox5 and Sox6 are required for notochord extracellular matrix sheath formation, notochord cell survival and formation of nuclei pulposi. Through these roles and essential roles in cartilage formation, they are central transcriptional regulators of vertebral column development. PMID- 12571106 TI - Combinatorial control of meristem identity in maize inflorescences. AB - The architecture of maize inflorescences, the male tassel and the female ear, is defined by a series of reiterative branching events. The inflorescence meristem initiates spikelet pair meristems. These in turn initiate spikelet meristems which finally produce the floret meristems. After initiating one meristem, the spikelet pair and spikelet meristem convert into spikelet and floret meristems, respectively. The phenotype of reversed germ orientation1 (rgo1) mutants is the production of an increased number of floret meristems by each spikelet meristem. The visible phenotypes include increased numbers of flowers in tassel and ear spikelets, disrupted rowing in the ear, fused kernels, and kernels with embryos facing the base of the ear, the opposite orientation observed in wild-type ears. rgo1 behaves as single recessive mutant. indeterminate spikelet1 (ids1) is an unlinked recessive mutant that has a similar phenotype to rgo1. Plants heterozygous for both rgo1 and ids1 exhibit nonallelic noncomplementation; these mutants fail to complement each other. Plants homozygous for both mutations have more severe phenotypes than either of the single mutants; the progression of meristem identities is retarded and sometimes even reversed. In addition, in rgo1; ids1 double mutants extra branching is observed in spikelet pair meristems, a meristem that is not affected by mutants of either gene individually. These data suggest a model for control of meristem identity and determinacy in which the progress through meristem identities is mediated by a dosage-sensitive pathway. This pathway is combinatorially controlled by at least two genes that have overlapping functions. PMID- 12571107 TI - A discrete transcriptional silencer in the bam gene determines asymmetric division of the Drosophila germline stem cell. AB - The Drosophila germline lineage depends on a complex microenvironment of extrinsic and intrinsic factors that regulate the self-renewing and asymmetric divisions of dedicated stem cells. Germline stem cells (GSCs) must express components of the Dpp cassette and the translational repressors Nanos and Pumilio, whereas cystoblasts require the bam and bgcn genes. Bam is especially attractive as a target of GSC differentiation factors because current evidence indicates that bam is both necessary and sufficient for cystoblast differentiation. In this paper, we have sought to distinguish between mutually exclusive transcriptional or post-transcriptional mechanisms as the primary regulators of bam expression in GSCs and cystoblasts. We find that bam transcription is active in young germ cells but is repressed specifically in GSCs. Activation depends on a 50 bp fragment that carries at least one germ cell specific enhancer element. A non-overlapping 18 bp sequence carries a transcriptional silencer that prevents bam expression in the GSC. Promoters lacking this silencer cause bam expression in the GSC and concomitant GSC loss. Thus, asymmetry of the GSC division can be reduced to identifying the mechanism that selectively activates the silencer element in GSCs. PMID- 12571109 TI - Cell contact-dependent positioning of the D cleavage plane restricts eye development in the Ilyanassa embryo. AB - In embryos of the gastropod Ilyanassa obsoleta, the first-quartet micromeres of the A, B and C lineages (1a, 1b, and 1c) are each competent to form an eye in response to signaling from the 3D cell. The first-quartet micromere of the dorsal D lineage (1d) is smaller than the others, divides at a slower rate, and lacks the ability to form an eye. These properties of 1d all depend on inheritance of vegetal polar lobe cytoplasm by its mother cell D at second cleavage. I show that they depend also on the presence of cells adjacent to D during the late four-cell stage: after ablation of the A and/or C cells before this stage, 1d inherits more cytoplasm than normal, divides more rapidly, and frequently forms an eye. In non D lineages, cleavage plane positioning and micromere division rates are relatively insensitive to cell contacts. Compressing whole embryos during third cleavage also leads to an increase in 1d volume correlated with abnormal eye formation; this suggests that the normal effect of cell contacts is to position the D cell cleavage furrow closer to the animal pole, and the enhanced division asymmetry of the D cell contributes to the suppression of eye development. PMID- 12571108 TI - Distal antenna and distal antenna related encode nuclear proteins containing pipsqueak motifs involved in antenna development in Drosophila. AB - Legs and antennae are considered to be homologous appendages. The fundamental patterning mechanisms that organize spatial pattern are conserved, yet appendages with very different morphology develop. A genetic hierarchy for specification of antennal identity has been partly elucidated. We report identification of a novel family of genes with roles in antennal development. The distal antenna (dan) and distal antenna-related (danr) genes encode novel nuclear proteins that are expressed in the presumptive distal antenna, but not in the leg imaginal disc. Ectopic expression of dan or danr causes partial transformation of distal leg structure toward antennal identity. Mutants that remove dan and danr activity cause partial transformation of antenna toward leg identity. Therefore we suggest that dan and danr contribute to differentiation of antenna-specific characteristics. Antenna-specific expression of dan and danr depends on a regulatory hierarchy involving homothorax and Distal-less, as well as cut and spineless. We propose that dan and danr are effector genes that act downstream of these genes to control differentiation of distal antennal structures. PMID- 12571110 TI - Targeted disruption of the homeobox gene Nkx2.9 reveals a role in development of the spinal accessory nerve. AB - The homeodomain-containing transcription factor Nkx2.9 is expressed in the ventralmost neural progenitor domain of the neural tube together with the related protein Nkx2.2 during early mouse embryogenesis. Cells within this region give rise to V3 interneurons and visceral motoneurons in spinal cord and hindbrain, respectively. To investigate the role of the Nkx2.9 gene, we generated a mutant mouse by targeted gene disruption. Homozygous mutant animals lacking Nkx2.9 were viable and fertile with no apparent morphological or behavioral phenotype. The distribution of neuronal progenitor cells and differentiated neurons in spinal cord was unaffected in Nkx2.9-deficient animals. This finding is in contrast to Nkx2.2-null mutants, which have been shown to exhibit ventral to dorsal transformation of neuronal cell fates in spinal cord. Our results suggest that specification of V3 interneurons in the posterior CNS does not require Nkx2.9, most probably because of functional redundancy with the co-expressed Nkx2.2 protein. In hindbrain, however, absence of Nkx2.9 resulted in a significantly altered morphology of the spinal accessory nerve (XIth), which appeared considerably shorter and thinner than in wild-type animals. Consistent with this phenotype, immature branchial motoneurons of the spinal accessory nerve, which normally migrate from a ventromedial to a dorsolateral position within the neural tube, were markedly reduced in Nkx2.9-deficient embryos at E10.5, while ventromedial motor column cells were increased in numbers. In addition, the vagal and glossopharyngeal nerves appeared abnormal in approximately 50% of mutant embryos, which may be related to the observed reduction of Phox2b expression in the nucleus ambiguus of adult mutant mice. From these observations, we conclude that Nkx2.9 has a specific function in the hindbrain as determinant of the branchial motoneuron precursor cells for the spinal accessory nerve and possibly other nerves of the branchial-motor column. Like other Nkx genes expressed in the CNS, Nkx2.9 seems to be involved in converting positional information into cell fate decisions. PMID- 12571111 TI - A mosaic genetic screen for genes necessary for Drosophila mushroom body neuronal morphogenesis. AB - Neurons undergo extensive morphogenesis during development. To systematically identify genes important for different aspects of neuronal morphogenesis, we performed a genetic screen using the MARCM system in the mushroom body (MB) neurons of the Drosophila brain. Mutations on the right arm of chromosome 2 (which contains approximately 20% of the Drosophila genome) were made homozygous in a small subset of uniquely labeled MB neurons. Independently mutagenized chromosomes (4600) were screened, yielding defects in neuroblast proliferation, cell size, membrane trafficking, and axon and dendrite morphogenesis. We report mutations that affect these different aspects of morphogenesis and phenotypically characterize a subset. We found that roadblock, which encodes a dynein light chain, exhibits reduced cell number in neuroblast clones, reduced dendritic complexity and defective axonal transport. These phenotypes are nearly identical to mutations in dynein heavy chain Dhc64 and in Lis1, the Drosophila homolog of human lissencephaly 1, reinforcing the role of the dynein complex in cell proliferation, dendritic morphogenesis and axonal transport. Phenotypic analysis of short stop/kakapo, which encodes a large cytoskeletal linker protein, reveals a novel function in regulating microtubule polarity in neurons. MB neurons mutant for flamingo, which encodes a seven transmembrane cadherin, extend processes beyond their wild-type dendritic territories. Overexpression of Flamingo results in axon retraction. Our results suggest that most genes involved in neuronal morphogenesis play multiple roles in different aspects of neural development, rather than performing a dedicated function limited to a specific process. PMID- 12571112 TI - Role of integrins in the development of the cerebral cortex. AB - Spatial and temporal changes in expression and function of integrin receptors in the developing cerebral wall parallel neurogenesis, radial glial differentiation, neuronal migration and the emergence of neuronal layers in the cerebral cortex. The distinct outcomes of integrin and extracellular matrix ligand mutations underscore the dynamic role they play in these processes during corticogenesis. The changing patterns of adhesive interactions mediated by integrins and their ligands across the cerebral wall during embryogenesis may set in motion developmental programs needed for progressive acquisition of different neuronal or glial phenotypes in the cerebral cortex. Here we discuss the role of integrins during cortical layer formation. PMID- 12571113 TI - The spatial pattern of response magnitude and selectivity for orientation and direction in cat visual cortex. AB - Optical imaging studies of orientation and direction preference in visual cortex have typically used vector averaging to obtain angle and magnitude maps. This method has shown half-rotation orientation singularities (pinwheels) located within regions of low orientation vector magnitude. Direction preference is generally orthogonal to orientation preference, but often deviates from this, particularly in regions of low direction vector magnitude. Linear regions of rapid change in direction preference terminate in or near orientation singularities. The vector-averaging method is problematic however because it does not clearly disambiguate spatial variation in orientation tuning width from variation in height. It may also wrongly estimate preferred direction in regions where preference is weak. In this paper we analyze optical maps of cat visual cortex by fitting model tuning functions to the responses. This new method reveals features not previously evident. Orientation tuning height and width vary independently across the map: tuning height is always low near singularities, however regions of broad and narrow orientation tuning width can be found in regions of low tuning height, often alternating in a spoke-like fashion around singularities. Orientation and direction preference angles are always closely orthogonal. Reversals in direction preference form lines that originate precisely in orientation singularities. PMID- 12571114 TI - Voltage-gated currents, dye and electrical coupling in the embryonic mouse neocortex. AB - We measured dye coupling, electrical coupling, and voltage-gated currents using whole-cell voltage clamp in slices of mouse sensorimotor cortex at embryonic day 14 (E14). As in rat ventricular zone (VZ), cells of the VZ were extensively dye coupled, often in clusters of >100 cells. In mouse VZ, however, cells were much less electrically coupled, making measurement of voltage-gated currents more accurate. All VZ cells expressed delayed K(+) currents (I(K)), and 30%, including morphologically identified radial glia, also expressed inward Na(+) currents (I(Na)). This fraction is consistent with I(Na) expression being an early event following cell cycle exit. Intermediate zone (IZ) cells also expressed I(K) and I(Na). Na(+) current amplitude distributions indicated three populations of IZ cells: those without I(Na), those with I(Na) similar in amplitude to VZ cells, and those with I(Na) being almost 10 times larger than in VZ cells. Cells of the cortical plate (CP) expressed both I(K) and I(Na), with I(Na) being almost 10 fold larger than in VZ cells. No cell in any zone expressed detectable hyperpolarization-activated currents. Our data suggest that the distribution and density of I(Na) may be related to early events of cell cycle exit and migration. PMID- 12571115 TI - Neural development of the neuregulin receptor ErbB4 in the cerebral cortex and the hippocampus: preferential expression by interneurons tangentially migrating from the ganglionic eminences. AB - The receptor tyrosine kinases represent an important class of signal transduction molecules that have been shown to play critical roles in neural development. We report in the present study that the neuregulin receptor ErbB4 is preferentially expressed by interneurons that are migrating tangentially from the ventral to the dorsal rat telencephalon. ErbB4 immunoreactivity was detected in the medial ganglionic eminence as early as embryonic day (E) 13 at the inception of tangential migration. Prominent ErbB4-positive migratory streams consisting of cells double-labeled with ErbB4 and Dlx, a marker of tangentially migrating cells, were found to advance along the lower intermediate zone and the marginal zone from the ventrolateral to the dorsomedial cortex at E16-E18. After E20, the ErbB4-positive stream in the lower intermediate zone shifted towards the germinal zone and further extended via the cortex into the hippocampal primordium. ErbB4 was not expressed by Tbr1-positive glutamatergic projection neurons during development. ErbB4 was preferentially expressed by the majority of parvalbumin positive interneurons and subsets of other GABAergic interneurons in the cerebral cortex and the hippocampus in adulthood. The early onset and preferential expression of ErbB4 in tangentially migrating interneurons suggests that neuregulin/ErbB4 signaling may regulate the development and function of telencephalic interneurons. PMID- 12571116 TI - FMRI evidence for an organization of prefrontal cortex by both type of process and type of information. AB - Neuroimaging evidence is conflicting regarding whether human prefrontal cortex (PFC) shows functional organization by type of processes engaged or type of information processed. Most studies use complex working or long-term memory tasks requiring multiple processes and the combinations of processes recruited for different materials may vary. Using functional magnetic resonance imaging (fMRI) and simple tasks suggested by a component process approach, we found activity in left PFC when participants thought about (refreshed) a just-seen item and in right PFC when participants noted whether an item had been presented previously. Furthermore, the distribution of activation in left or right PFC varied with type of information. Thus, at the component process level, PFC shows functional organization by both process and type of information. PMID- 12571117 TI - Timing and precision of spike initiation in layer V pyramidal cells of the rat somatosensory cortex. AB - The timing and location of action potential initiation in layer V pyramidal cells of the rat somatosensory cortex depends on different ligand-gated and intrinsic conductances. In order to assess the shunting effect of GABA(A) receptor activation and I(h) on the summative properties of these neurons, we recorded simultaneously from their somata and dendrites and induced combined excitatory and inhibitory postsynaptic potentials using extracellular stimulation. Three major consequences of GABA(A) receptor activation on the integration properties were found: (i) the action potential could be initiated in the distal apical dendrite, if GABA(A) receptors were simultaneously activated in the soma; (ii) GABA(A) receptor activation narrowed the time window for suprathreshold summation of two coincident excitatory synaptic inputs by a factor of 3.4; and (iii) uneven weights of the GABAergic components between two inputs could lead to unbalanced shunting and consequently to a time shift in the peak of the coincidence detection window. Blockade of I(h) practically abolished coincidence detection within a time window of 30 ms. Location of spike initiation and summation of coincident inputs in pyramidal cells is determined by at least two shunting mechanisms: activation of the presynaptic inhibitory network and modulation of I(h) as a function of the postsynaptic membrane potential. PMID- 12571118 TI - Long-term gonadectomy affects the density of tyrosine hydroxylase- but not dopamine-beta-hydroxylase-, choline acetyltransferase- or serotonin immunoreactive axons in the medial prefrontal cortices of adult male rats. AB - The rat prefrontal cortices participate in cognitive, affective and mnemonic functions. The importance of dopamine innervation for these computations is illustrated in studies showing that both supranormal levels and chemical lesions of prefrontal dopamine induce severe behavioral deficits. Observed hormone effects on some of these same behaviors suggest that the prefrontal cortices are also sensitive to gonadal steroids. These two influences seem to converge in recent evidence of increased dopamine axon density in representative prefrontal but not sensory or motor cortices in gonadectomized adult male rats. The seeming selectivity of these effects was further explored here using immunocytochemistry for tyrosine hydroxylase, dopamine-b-hydroxylase, serotonin and choline acetyltransferase to label neurochemically identified afferents in remaining, unstudied prefrontal fields of rat cortex in animals that were sham-operated or gonadectomized and given placebo, testosterone propionate, estradiol or dihydrotestosterone 28 days before being killed. Group comparisons revealed that across prefrontal zones, gonadectomy produced androgen-sensitive increases in presumed dopamine axon density, but did not affect the other afferents. These findings thus bolster evidence for a targeted gonadal steroid influence involving the prefrontal cortices and a neurotransmitter essential for their normal operations and implicated in their dysfunction in disorders such as schizophrenia as well. PMID- 12571119 TI - Different populations of tyrosine-hydroxylase-immunoreactive neurons defined by differential expression of nitric oxide synthase in the human temporal cortex. AB - In the mammalian neocortex, neurons containing tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis, constitute an enigmatic and ill defined group of aspiny non-pyramidal cells. In the human neocortex, these neurons are mostly found in layers V-VI, the same layers in which another conspicuous group of nitrergic non-pyramidal cells are found - those containing nitric oxide synthase (nNOS) and that can be labeled by nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) histochemistry. The main aim of the present study was to determine the extent to which neurons and fibers containing TH, NADPHd or nNOS co-localize in the human temporal cortex, using immunocytochemistry and NADPHd histochemistry. Furthermore, we have quantified the degree to which axons immunoreactive (ir) for TH contact the somata of neurons by co-labeling with the neuron-specific nuclear protein NeuN. As a result, we show that the population of TH-ir neurons can be subdivided into two main neurochemical groups: those expressing nNOS (26%) and those that do not (74%). There was no co-localization of TH with nNOS in the prominent horizontally oriented plexus of fibers in layer I and we did not observe any double bouquet cells, chandelier cells or basket cells that contained TH. Finally, we observed that only 6% of the TH-ir axonal boutons examined (n = 1724) could be seen to contact neuronal somata. Thus, most TH-ir axons must form synapses with dendrites. In conjunction with data from previous studies, these results suggest that TH is found in different neurochemically defined subpopulations of non pyramidal neurons in layers V-VI of the human temporal cortex. Consequently, it appears that a partial overlap of the catecholaminergic and nitrergic systems is probably due to the intrinsic cortical TH-nNOS-ir neurons. PMID- 12571120 TI - Representations of pleasant and painful touch in the human orbitofrontal and cingulate cortices. AB - The cortical areas that represent affectively positive and negative aspects of touch were investigated using functional magnetic resonance imaging (fMRI) by comparing activations produced by pleasant touch, painful touch produced by a stylus, and neutral touch, to the left hand. It was found that regions of the orbitofrontal cortex were activated more by pleasant touch and by painful stimuli than by neutral touch and that different areas of the orbitofrontal cortex were activated by the pleasant and painful touches. The orbitofrontal cortex activation was related to the affective aspects of the touch, in that the somatosensory cortex (SI) was less activated by the pleasant and painful stimuli than by the neutral stimuli. This dissociation was highly significant for both the pleasant touch (P < 0.006) and for the painful stimulus (P < 0.02). Further, it was found that a rostral part of the anterior cingulate cortex was activated by the pleasant stimulus and that a more posterior and dorsal part was activated by the painful stimulus. Regions of the somatosensory cortex, including SI and part of SII in the mid-insula, were activated more by the neutral touch than by the pleasant and painful stimuli. Part of the posterior insula was activated only in the pain condition and different parts of the brainstem, including the central grey, were activated in the pain, pleasant and neutral touch conditions. The results provide evidence that different areas of the human orbitofrontal cortex are involved in representing both pleasant touch and pain, and that dissociable parts of the cingulate cortex are involved in representing pleasant touch and pain. PMID- 12571121 TI - Instructed delay activity in the human prefrontal cortex is modulated by monetary reward expectation. AB - Goal-directed actions are executed with greater efficiency when the goals of the actions are rewarded and so the reward expectation must influence systems concerned with action-planning and motor control. However, little is known about how this influence is achieved in primates. Here, we demonstrate in human subjects that manual performance is enhanced when the goals of the visually cued actions are monetary rewards. We also used event-related fMRI in the same subjects to localize neural activity related to action preparation and selection that was influenced by the reward. We found three areas with significant interaction between reward and preparation: the prestriate visual cortex, the premotor cortex and the lateral prefrontal cortex. The latter two areas appear to be frontal systems integrating the expectation of rewards with selection and preparation of actions. PMID- 12571122 TI - Syncope, cerebral perfusion, and oxygenation. AB - During standing, both the position of the cerebral circulation and the reductions in mean arterial pressure (MAP) and cardiac output challenge cerebral autoregulatory (CA) mechanisms. Syncope is most often associated with the upright position and can be provoked by any condition that jeopardizes cerebral blood flow (CBF) and regional cerebral tissue oxygenation (cO(2)Hb). Reflex (vasovagal) responses, cardiac arrhythmias, and autonomic failure are common causes. An important defense against a critical reduction in the central blood volume is that of muscle activity ("the muscle pump"), and if it is not applied even normal humans faint. Continuous tracking of CBF by transcranial Doppler-determined cerebral blood velocity (V(mean)) and near-infrared spectroscopy-determined cO(2)Hb contribute to understanding the cerebrovascular adjustments to postural stress; e.g., MAP does not necessarily reflect the cerebrovascular phenomena associated with (pre)syncope. CA may be interpreted as a frequency-dependent phenomenon with attenuated transfer of oscillations in MAP to V(mean) at low frequencies. The clinical implication is that CA does not respond to rapid changes in MAP; e.g., there is a transient fall in V(mean) on standing up and therefore a feeling of lightheadedness that even healthy humans sometimes experience. In subjects with recurrent vasovagal syncope, dynamic CA seems not different from that of healthy controls even during the last minutes before the syncope. Redistribution of cardiac output may affect cerebral perfusion by increased cerebral vascular resistance, supporting the view that cerebral perfusion depends on arterial inflow pressure provided that there is a sufficient cardiac output. PMID- 12571123 TI - Microvascular gas embolization clearance following perfluorocarbon administration. AB - Effective treatment of vascular gas embolism may be possible with emulsified fluorocarbon compounds. We tested the hypothesis that a fluorocarbon emulsion delivered before gas embolization would enhance bubble motion through the vasculature, favoring more rapid clearance. Air microbubbles were injected into the rat cremaster microcirculation in six groups of rats receiving Perftoran, an emulsified fluorocarbon, or saline immediately before, 2 h before, or after bubble injection. Embolism dimensions and dynamics were observed by using intravital microscopy. Surface area at lodging was equal between groups. Bubbles having smaller volume embolized smaller diameter vessels in the Perftoran pretreatment groups. A higher incidence of bubble dislodgement and larger distal displacement occurred in these two groups, with a 36% decrease in the time to bubble clearance and restoration of blood flow. Intravascular emulsified fluorocarbon administration before gas embolization affected initial bubble conformation, increased bubble dislodgement, and resulted in bubble displacement further into the periphery of the microcirculation. These dynamic events did not occur if embolization preceded fluorocarbon administration. PMID- 12571124 TI - Retinal venous oxygen saturation and cardiac output during controlled hemorrhage and resuscitation. AB - The objective was to test calibration of an eye oximeter (EOX) in a vitiligo swine eye and correlate retinal venous oxygen saturation (Srv(O(2))), mixed venous oxygen saturation (Sv(O(2))), and cardiac output (CO) during robust changes in blood volume. Ten anesthetized adult Sinclair swine with retinal vitiligo were placed on stepwise decreasing amounts of oxygen. At each oxygen level, femoral artery oxygen saturation (Sa(O(2))) and retinal artery oxygen saturation (Sra(O(2))) were obtained. After equilibration on 100% O(2), subjects were bled at 1.4 ml. kg(-1). min(-1) for 20 min. Subsequently, anticoagulated shed blood was reinfused at the same rate. During graded hypoxia, exsanguination, and reinfusion, Sra(O(2)) and Srv(O(2)) were measured by using the EOX, and CO and Sv(O(2)) were measured by using a pulmonary artery catheter. During graded hypoxia, Sra(O(2)) correlated with Sa(O(2)) (r = 0.92). Srv(O(2)) correlated with Sv(O(2)) (r = 0.89) during exsanguination and reinfusion. Sv(O(2)) and Srv(O(2)) correlated with CO during blood removal and resuscitation (r = 0.92). Use of vitiligo retinas improved the calibration of EOX measurements. In this robust hemorrhage model, Srv(O(2)) correlates with CO and Sv(O(2)) across the range of exsanguination and resuscitation. PMID- 12571125 TI - Aging and the time and frequency structure of force output variability. AB - The present study examined the time and frequency structure of force output in adult humans to determine whether the changes in complexity with age are dependent on external task demands. Healthy young (20-24 yr), old (60-69 yr), and older-old (75-90 yr) humans produced isometric force contractions to constant and sine wave targets that also varied in force level. First, force variability on each force task increased with advancing age. Second, both time and frequency analysis showed that the structure of the force output in the old and older-old adults was less complex in the constant-force level task and more complex in the sine wave force task. Third, the alterations in force output with aging were primarily due to low-frequency bands <4 Hz. These results support the postulation that the observed increase or decrease in physiological complexity with aging is influenced by the relatively fast time scale of external task demands (Vaillancourt DE and Newell KM. Neurobiol Aging 23: 1-11, 2002). PMID- 12571126 TI - Respiratory response to activation or disinhibition of the dorsal periaqueductal gray in rats. AB - The neural substrates mediating autonomic components of the behavioral defense response have been shown to reside in the periaqueductal gray (PAG). The cardiovascular components of the behavioral defense response have been well described and are tonically suppressed by GABAergic input. The ventilatory response associated with disinhibition of the dorsal PAG (dPAG) neurons is unknown. In urethane-anesthetized, spontaneously breathing rats, electrical stimulation of the dPAG was shown to decrease the expiration time and increase respiratory frequency, with no change in time of inspiration. Baseline and the change in diaphragm electromyograph also increased, resulting in an increase in neural minute activity. Microinjection of bicuculline methobromide, a GABA(A) receptor antagonist, into the dPAG produced a similar response, which was dose dependent. Disinhibition of the dPAG also produced a decrease in inspiration time. These results suggest that GABA(A)-mediated suppression of dPAG neurons plays a role in the respiratory component of behavioral defense responses. The respiratory change is due in part to a change in brain stem respiratory timing and phasic inspiratory output. In addition, there is an increase in tonic diaphragm activity. PMID- 12571127 TI - Endurance training-induced changes in alkali light chain patterns in type IIB fibers of the rat. AB - The effects of endurance training on the expression of myosin were electrophoretically analyzed in the deep portion of vastus lateralis muscle from the rat. A 10-wk running program led to increases (P < 0.01) in myosin heavy chain (MHC) 2a and 2d with a decrease (P < 0.01) in MHC(2b). Training also evoked a rearrangement of the isomyosin pattern with decreases in fast isomyosin (FM) 1 (P < 0.01) and FM2 (P < 0.05) and a rise in intermediate isomyosin (P < 0.01). These changes were accompanied by a 61% decrease (P < 0.01) in myosin light chain (MLC) 3F (11.8 +/- 2.7 vs. 4.6 +/- 4.2%). Two-dimensional electrophoresis made it possible to separate the triplet of isomyosins (FMb) consisting of MHC(2b). Training elicited a 26% decrease (P < 0.05) in the FM1b fraction within FMb, i.e., FM1b/(FM1b + FM2b + FM3b) (24.2 +/- 5.5 vs. 18.0 +/- 4.3%). These changes resulted in a 10% decrease (P < 0.05) in the MLC(3F) fraction, i.e., MLC(3F)/(MLC(1F) + MLC(3F)), in FMb (44.9 +/- 4.5 vs. 40.3 +/- 3.2%). These results suggest that endurance training may exert the depressive effect on the contractile velocity of type IIB fibers and that a training-induced decrease in the contractile velocity of whole muscle may be caused by alterations in fast alkali MLC complements within a given fiber type as well as by transitions in MHC based fiber populations. PMID- 12571128 TI - Cutaneous vasoconstrictor response to whole body skin cooling is altered by time of day. AB - To test for a diurnal difference in the vasoconstrictor control of the cutaneous circulation, we performed whole body skin cooling (water-perfused suits) at 0600 (AM) and 1600 (PM). After whole body skin temperature (T(sk)) was controlled at 35 degrees C for 10 min, it was progressively lowered to 32 degrees C over 18-20 min. Skin blood flow (SkBF) was monitored by laser-Doppler flowmetry at three control sites and at a site that had been pretreated with bretylium by iontophoresis to block noradrenergic vasoconstriction. After whole body skin cooling, maximal cutaneous vascular conductance (CVC) was measured by locally warming the sites of SkBF measurement to 42 degrees C for 30 min. Before whole body skin cooling, sublingual temperature (T(or)) in the PM was significantly higher than that in the AM (P < 0.05), but CVC, expressed as a percentage of maximal CVC (%CVC(max)), was not statistically different between AM and PM. During whole body skin cooling, %CVC(max) levels at bretylium-treated sites in AM or PM were not significantly reduced from baseline. In the PM, %CVC(max) at control sites fell significantly at T(sk) of 34.3 +/- 0.01 degrees C and lower (P < 0.05). In contrast, in the AM %CVC(max) at control sites was not significantly reduced from baseline until T(sk) reached 32.3 +/- 0.01 degrees C and lower (P < 0.05). Furthermore, the decrease in %CVC(max) in the PM was significantly greater than that in AM at T(sk) of 33.3 +/- 0.01 degrees C and lower (P < 0.05). Integrative analysis of the CVC response with respect to both T(or) and T(sk) showed that the cutaneous vasoconstrictor response was shifted to higher internal temperatures in the PM. These findings suggest that during whole body skin cooling the reflex control of the cutaneous vasoconstrictor system is shifted to a higher internal temperature in the PM. Furthermore, the slope of the relationship between CVC and T(sk) is steeper in the PM compared with that in the AM. PMID- 12571129 TI - High oxygen concentrations predispose mouse lungs to the deleterious effects of high stretch ventilation. AB - Mechanical ventilation is a necessary intervention for patients with acute lung injury. However, mechanical ventilation can propagate acute lung injury and increase systemic inflammation. The exposure to >21% oxygen is often associated with mechanical ventilation yet has not been examined within the context of lung stretch. We hypothesized that mice exposed to >90% oxygen will be more susceptible to the deleterious effects of high stretch mechanical ventilation. C57B1/6 mice were randomized into 48-h exposure of 21 or >90% oxygen; mice were then killed, and isolated lungs were randomized into a nonstretch or an ex vivo, high-stretch mechanical ventilation group. Lungs were assessed for compliance and lavaged for surfactant analysis, and cytokine measurements or lungs were homogenized for surfactant-associated protein analysis. Mice exposed to >90% oxygen + stretch had significantly lower compliance, altered pulmonary surfactant, and increased inflammatory cytokines compared with all other groups. Our conclusion is that 48 h of >90% oxygen and high-stretch mechanical ventilation deleteriously affect lung function to a greater degree than stretch alone. PMID- 12571130 TI - Effect of quadriceps femoris muscle length on neural activation during isometric and concentric contractions. AB - The effect of muscle length on neural drive (here termed "neural activation") was investigated from electromyographic activities and activation levels (twitch interpolation). The neural activation was measured in nine men during isometric and concentric (30 and 120 degrees /s) knee extensions for three muscle lengths (35, 55, and 75 degrees knee flexion, i.e., shortened, intermediate, and lengthened muscles, respectively). Long (76 degrees ), medium (56 degrees ), and short (36 degrees ) ranges of motion were used to investigate the effect of the duration of concentric contraction. Neural activation was found to depend on muscle length. Reducing the duration of contraction had no effect. Neural activation was higher with short muscle length during isometric contractions and was weaker for shortened than for intermediate and lengthened muscles performing 120 degrees /s concentric contractions. Muscle length had no effect on 30 degrees /s concentric neural activation. Peripheral mechanisms and discharge properties of the motoneurons could partly explain the observed differences in the muscle length effect. We thus conclude that muscle length has a predominant effect on neural activation that would modulate the angular velocity dependency. PMID- 12571131 TI - Impedance, gas mixing, and bimodal ventilation in constricted lungs. AB - To evaluate the effect of increasing smooth muscle activation on the distribution of ventilation, lung impedance and expired gas concentrations were measured during a 16-breath He-washin maneuver in five nonasthmatic subjects at baseline and after each of three doses of aerosolized methacholine. Values of dynamic lung elastance (El,dyn), the curvature of washin plots, and the normalized slope of phase III (S(N)) were obtained. At the highest dose, El,dyn was 2.6 times the control value and S(N) for the 16th breath was 0.65 liter(-1). A previously described model of a constricted terminal airway was extended to include variable muscle activation, and the extended model was tested against these data. The model predicts that the constricted airway has two stable states. The impedances of the two stable states are independent of smooth muscle activation, but driving pressure and the number of airways in the high-resistance state increase with increasing muscle activation. Model predictions and experimental data agree well. We conclude that, as a result of the bistability of the terminal airways, the ventilation distribution in the constricted lung is bimodal. PMID- 12571132 TI - A comparison of voluntary and electrically induced contractions by interleaved 1H and 31P-NMRS in humans. AB - Skeletal muscle voluntary contractions (VC) and electrical stimulations (ES) were compared in eight healthy men. High-energy phosphates and myoglobin oxygenation were simultaneously monitored in the quadriceps by interleaved (1)H- and (31)P NMR spectroscopy. For the VC protocol, subjects performed five or six bouts of 5 min with a workload increment of 10% of maximal voluntary torque (MVT) at each step. The ES protocol consisted of a 13-min exercise with a load corresponding to 10% MVT. For both protocols, exercise consisted of 6-s isometric contractions and 6-s rest cycles. For an identical mechanical level (10% MVT), ES induced larger changes than VC in the P(i)-to-phosphocreatine ratio [1.38 +/- 1.14 (ES) vs. 0.13 +/- 0.04 (VC)], pH [6.69 +/- 0.11 (ES) vs. 7.04 +/- 0.07 (VC)] and myoglobin desaturation [43 +/- 15.9 (ES) vs. 6.1 +/- 4.6% (VC)]. ES activated the muscle facing the NMR coil to a greater extent than did VCs when evaluated under identical technical conditions. This metabolic pattern can be interpreted in terms of specific temporal and spatial muscle cell recruitment. Furthermore, at identical levels of energy charge, the muscle was more acidotic and cytoplasm appeared more oxygenated during ES than during VC. These results are in accordance with a preferential recruitment of type II fibers and a relative muscle hyperperfusion during ES. PMID- 12571133 TI - Antioxidants attenuate the plasma cytokine response to exercise in humans. AB - Exercise increases plasma TNF-alpha, IL-1beta, and IL-6, yet the stimuli and sources of TNF-alpha and IL-1beta remain largely unknown. We tested the role of oxidative stress and the potential contribution of monocytes in this cytokine (especially IL-1beta) response in previously untrained individuals. Six healthy nonathletes performed two 45-min bicycle exercise sessions at 70% of Vo(2 max) before and after a combination of antioxidants (vitamins E, A, and C for 60 days; allopurinol for 15 days; and N-acetylcysteine for 3 days). Blood was drawn at baseline, end-exercise, and 30 and 120 min postexercise. Plasma cytokines were determined by ELISA and monocyte intracellular cytokine level by flow cytometry. Before antioxidants, TNF-alpha increased by 60%, IL-1beta by threefold, and IL-6 by sixfold secondary to exercise (P < 0.05). After antioxidants, plasma IL-1beta became undetectable, the TNF-alpha response to exercise was abolished, and the IL 6 response was significantly blunted (P < 0.05). Exercise did not increase the percentage of monocytes producing the cytokines or their mean fluorescence intensity. We conclude that in untrained humans oxidative stress is a major stimulus for exercise-induced cytokine production and that monocytes play no role in this process. PMID- 12571134 TI - Reduced exercise arteriovenous O2 difference in Type 2 diabetes. AB - Maximal O(2) consumption (Vo(2 max)) is lower in individuals with Type 2 diabetes than in sedentary nondiabetic individuals. This study aimed to determine whether the lower Vo(2 max) in diabetic patients was due to a reduction in maximal cardiac output (Q(max)) and/or peripheral O(2) extraction. After 11 Type 2 diabetic patients and 12 nondiabetic subjects, matched for age and body composition, who had not exercised for 2 yr, performed a bicycle ergometer exercise test to determine Vo(2 max), submaximal cardiac output, Q(max), and arterial-mixed venous O(2) (a-v O(2)) difference were assessed. Maximal workload, Vo(2 max), and maximal a-v O(2) difference were lower in Type 2 diabetic patients (P < 0.05). Q(max) was low in both groups but not significantly different: 11.2 and 10.0 l/min for controls and diabetic patients, respectively (P > 0.05). Submaximal O(2) uptake and heart rate were lower at several workloads in diabetic patients; respiratory exchange ratio was similar between groups at all workloads. Vo(2 max) was linearly correlated with a-v O(2) difference, but not Q(max) in diabetic patients. These data suggest that a reduction in maximal a-v O(2) difference contributes to a decreased Vo(2 max) in Type 2 diabetic patients. PMID- 12571135 TI - Temporary tracheal occlusion in fetal sheep with lung hypoplasia does not improve postnatal lung function. AB - Prolonged fetal tracheal occlusion (TO) accelerates lung growth but leads to loss of alveolar epithelial type II (AE2) cells. In contrast, temporary TO leads to recovery of AE2 cells and their ability to produce surfactant. The aim of this study was to determine the effects of temporary TO in fetal sheep with lung hypoplasia on postnatal lung function, structure, and surfactant protein mRNA expression. Diaphragmatic hernia (DH) was created in 22 fetal sheep at 65 days of gestation. TO was performed between 110 days of gestation and full term (DH/TO, n = 7) and between 110 and 130 days of gestation (DH/TO+R, n = 6). Sham-operated fetuses (n = 11) served as controls. Lambs were delivered at approximately 139 days of gestation, and blood gas tensions were monitored over a 2-h resuscitation period. Temporary TO increased growth of the hypoplastic lung and restored surfactant protein mRNA expression and AE2 cell density but did not improve respiratory function above that of animals that underwent prolonged TO; DH/TO and DH/TO+R lambs were hypoxic and hypercapnic compared with Sham animals. Lung compliance remained low in DH/TO+R lambs, most likely as a consequence of the persistent increase in alveolar wall thickness in these animals. PMID- 12571136 TI - Physiological, metabolic, and performance implications of a prolonged hill walk: influence of energy intake. AB - We aimed to examine the effects of different energy intakes on a range of responses that are relevant to the safety of hill walkers. In a balanced design, 16 men completed a strenuous self-paced mountainous hill walk over 21 km, under either a low-energy (2.6 MJ; 616 kcal) intake (LEI) or high-energy (12.7 MJ; 3,019 kcal) intake (HEI) condition. During the hill walk, rectal temperatures were measured continuously, and blood samples for the analysis of metabolites and hormones were drawn before breakfast and immediately after the walk. Subjects also completed a battery of performance tests that included muscular strength, reaction times, flexibility, balance, and kinesthetic differentiation tests. During the LEI, mean blood glucose concentrations leveled off at the low-middle range of normoglycemia, whereas, on the HEI, they were significantly elevated compared with the LEI. The maintained blood glucose concentrations, during the LEI, were probably mediated via the marked fat mobilization, reflected by a two- to fivefold increase in nonesterified fatty acids, 3-hydroxybutyrate, and glycerol concentrations. The LEI group showed significantly slower one- and two finger reaction time, had an impaired ability to balance, and were compromised in their ability to maintain body temperature, when compared with the HEI group. The modestly impaired performance (particularly with respect to balance) and thermoregulation during the LEI condition may increase susceptibility to both fatigue and injury during the pursuit of recreational activity outdoors. PMID- 12571137 TI - Breath-by-breath measurement of the volume displaced by diaphragm motion. AB - To develop an accurate method to measure the volume displaced by diaphragm motion (DeltaVdi) breath by breath, we compared DeltaVdi measured by a previously evaluated biplanar radiographic method (Singh B, Eastwood PR, and Finucane KE. J Appl Physiol 91: 1913-1923, 2001) at several lung volumes during vital capacity inspirations in 10 healthy and nine hyperinflated subjects with 1) DeltaVdi measured from the same chest X-rays by two previously described uniplanar methods (Petroll WM, Knight H, and Rochester DF. J Appl Physiol 69: 2175-2182, 1990; Verschakelen JA, Deschepper K, and Demendts M. J Appl Physiol 72: 1536-1540, 1992) and a proposed method that considered actual cross-sectional shape of the rib cage and spinal volume (DeltaVdi(S)); and 2) DeltaVdi(S) measured by lateral fluoroscopy in the same 10 healthy subjects. Relative to biplanar DeltaVdi, DeltaVdi(S) values from lateral chest X-rays and fluoroscopy were not different, whereas DeltaVdi values of Petroll et al. and Verschakelen et al. were increased by (means +/- SD) 1.98 +/- 1.59 and 1.16 +/- 0.82 liters, respectively (both P < 0.001). During quiet breathing, DeltaVdi(S) by lateral fluoroscopy was 66 +/- 16% of tidal volume and similar to that between functional residual capacity and one half inspiratory capacity by the biplanar radiographic method. We conclude that accurate breath-by-breath measurements of DeltaVdi can be made by using lateral fluoroscopy. PMID- 12571138 TI - Myofascial force transmission: muscle relative position and length determine agonist and synergist muscle force. AB - Equal proximal and distal lengthening of rat extensor digitorum longus (EDL) were studied. Tibialis anterior, extensor hallucis longus, and EDL were active maximally. The connective tissues around these muscle bellies were left intact. Proximal EDL forces differed from distal forces, indicating myofascial force transmission to structures other than the tendons. Higher EDL distal force was exerted (ratio approximately 118%) after distal than after equal proximal lengthening. For proximal force, the reverse occurred (ratio approximately 157%). Passive EDL force exerted at the lengthened end was 7-10 times the force exerted at the nonlengthened end. While kept at constant length, synergists (tibialis anterior + extensor hallucis longus: active muscle force difference approximately -10%) significantly decreased in force by distal EDL lengthening, but not by proximal EDL lengthening. We conclude that force exerted at the tendon at the lengthened end of a muscle is higher because of the extra load imposed by myofascial force transmission on parts of the muscle belly. This is mediated by changes of the relative position of most parts of the lengthened muscle with respect to neighboring muscles and to compartment connective tissues. As a consequence, muscle relative position is a major codeterminant of muscle force for muscle with connectivity of its belly close to in vivo conditions. PMID- 12571139 TI - Carotid body denervation effect on cytochrome oxidase activity in pre-Botzinger complex of developing rats. AB - Previously, we found that the rat pre-Botzinger complex (PBC) exhibited reduced cytochrome oxidase (CO) activity on postnatal days (P) 3-4 and especially on P12, with a concomitant decrease in glutamate and N-methyl-d-aspartate receptor subunit 1, and an increase in GABA, GABA(B), glycine receptor, and glutamate subunit 2. We hypothesized that the PBC would be more affected by carotid body denervation (CBD) during the two critical windows than at other times. Pairs of CBD and sham animals at each postnatal day from P2 to P14 and at P21 were operated on and survived for 3 days. Brain stems were processed for CO and neurokinin-1 receptor for the identification of PBC. Results indicate that CBD caused a significant loss in body weight in all animals and a reduction in PBC somal size when the surgery was between P2 and P7. CBD also induced a significant decrease in CO activity of the PBC in most animals and a distinct delay, as well as prolongation of the maturational process, especially when induced close to P3 and P11-P13. PMID- 12571140 TI - Effect of somatic growth, strain, and sex on double-chamber plethysmographic respiratory function values in healthy mice. AB - Double-chamber plethysmography has been recognized since 1979 as a reference technique to measure pulmonary function values in guinea pigs, but it has not gained attention for use in mice. Theoretically, however, this technique combines the advantages of single-chamber plethysmography with a quantitative assessment of flow and/or volume and a calculated resistance, the interpretation of which in terms of bronchoconstriction is not disputed. Here we show that, when appropriately preconditioned, mice are able to gradually grow accustomed to the apparatus and display extremely stable nasal and thoracoabdominal flow tracings. Overall, strain, sex, and somatic growth had a significant effect on pulmonary function values. The changes in specific airway resistance (sRaw) and enhanced pause (Penh) values were never in the same direction, indicating that they measure different things. The respiratory frequency was far higher in C57BL/6 compared with BALB/c mice. Peak flows, minute volume, specific tidal and minute volumes, and sRaw were also higher, but Penh was smaller. Males breathed at a higher frequency than females, leading to a higher minute volume. Nevertheless, the specific volumes were considerably higher among females. Penh was lower in males, whereas sRaw was identical in both sexes. Changes associated with somatic growth were rapid and important between 5 and 9 wk, then slowed down between 9 and 12-13 wk and became almost imperceptible after. PMID- 12571141 TI - Overload-induced androgen receptor expression in the aged rat hindlimb receiving nandrolone decanoate. AB - This study's purpose was to examine whether functional overload with nandrolone decanoate (ND) administration increased muscle mass and steroid receptor concentration in aged rat soleus (Sol) and plantaris (Plan) muscle. ND (6 mg/kg body wt) was administered once a week for 4 wk, whereas control rats received sesame seed oil injections. Functional overload of the hindlimb Sol and Plan was induced by synergistic gastrocnemius muscle ablation at the beginning of the fourth week. Adult (5 mo of age) and aged rats (25 mo of age) were randomly assigned to four groups: control, overload, control-ND, and overload-ND. Seven days of functional overload increased adult Sol muscle mass 27%, whereas the aged Sol muscle mass did not change. The aged overloaded Sol muscle receiving ND significantly increased muscle weight by 35% and total muscle protein by 24%. Aged Plan muscle did not increase muscle weight with overload or ND treatment. Androgen receptor protein was induced by ND treatment and functional Ov, and combining the two treatments induced Sol androgen receptor protein concentration above either alone. Sol glucocorticoid receptor protein concentration increased in overload groups of both ages. ND administration can increase aged Sol muscle mass and protein content after 7 days of functional overload, and the cooperative induction of androgen receptor may be important for this response. PMID- 12571142 TI - Effect of myocardial volume overload and heart failure on lactate transport into isolated cardiac myocytes. AB - The purpose of this study was to determine lactate transport kinetics in single isolated rat ventricular cardiac myocytes after 1) 8 wk of myocardial volume overload (MVO) and 2) congestive heart failure (CHF). Twenty male Sprague-Dawley rats were assigned to one of four groups: myocardial hypertrophy (MH), MH sham (MHS), CHF, or CHF sham (CHFS). A chronic MVO was induced in the MH and CHF groups by an infrarenal arteriovenous fistula. Postdeath heart and lung weights were significantly greater (P < 0.05) for the MH and CHF groups compared with controls. Isolated cardiac myocytes were loaded with BCECF to determine intracellular pH (pH(i)) changes after the addition of lactate to the extracellular superfusate. Alterations in pH(i) with the addition of varied lactate concentrations were attenuated 72-89% by 5.0 mM alpha-cyano-4 hydroxycinnamate. Significant differences (P < 0.05) were found in estimated maximal lactate transport rates between the experimental and sham groups (MH = 19.4 +/- 1.1 nmol x microl(-1) x min(-1) vs. MHS = 15.1 +/- 1.1 nmol x microl(-1) x min(-1); CHF = 20.2 +/- 2.0 nmol x microl(-1) x min(-1) vs. CHFS = 14.0 +/- 0.9 nmol x microl(-1) x min(-1)). Western blot analysis confirmed a 270% increase in monocarboxylate symport protein 1 (MCT1) protein content in CHF compared with CHFS rats. The results of this study suggest that MH and CHF induced by MVO engender a greater maximal lactate transport capacity across the cardiac myocyte sarcolemma along with an increase in MCT1 protein content. These alterations would likely benefit the cell by attenuating intracellular acidification during a period of increased myocardial load. PMID- 12571143 TI - Central CO2 chemoreception in developing bullfrogs: anomalous response to acetazolamide. AB - Central CO(2) chemoreception and the role of carbonic anhydrase were assessed in brain stems from Rana catesbeiana tadpoles and frogs. Buccal and lung rhythms were recorded from cranial nerve VII and spinal nerve II during normocapnia and hypercapnia before and after treatment with 25 microM acetazolamide. The lung response to acetazolamide mimicked the hypercapnic response in early-stage and midstage metamorphic tadpoles and frogs. In late-stage tadpoles, acetazolamide actually inhibited hypercapnic responses. Acetazolamide and hypercapnia decreased the buccal frequency but had no effect on the buccal duty cycle. Carbonic anhydrase activity was present in the brain stem in every developmental stage. Thus more frequent lung ventilation and concomitantly less frequent buccal ventilation comprised the hypercapnic response, but the response to acetazolamide was not consistent during metamorphosis. Therefore, acetazolamide is not a useful tool for central CO(2) chemoreceptor studies in this species. The reversal of the effect of acetazolamide in late-stage metamorphosis may reflect reorganization of central chemosensory processes during the final transition from aquatic to aerial respiration. PMID- 12571144 TI - Invited review: Mechanisms underlying motor unit plasticity in the respiratory system. AB - Neuromotor control of skeletal muscles, including respiratory muscles, is ultimately dependent on the function of the motor unit (comprising an individual motoneuron and the muscle fibers it innervates). Considerable diversity exists across diaphragm motor units, yet remarkable homogeneity is present (and maintained) within motor units. In recent years, the mechanisms underlying the development and adaptability of respiratory motor units have received great attention, leading to significant advances in our understanding of diaphragm motor unit plasticity. For example, following imposed inactivity of the diaphragm muscle, there are changes at phrenic motoneurons, neuromuscular junctions, and muscle fibers that tend to restore the ability of the diaphragm to sustain ventilation. The role of activity, neurotrophins, and other growth factors in modulating this adaptability is discussed. PMID- 12571145 TI - Invited review: Neural network plasticity in respiratory control. AB - Respiratory network plasticity is a modification in respiratory control that persists longer than the stimuli that evoke it or that changes the behavior produced by the network. Different durations and patterns of hypoxia can induce different types of respiratory memories. Lateral pontine neurons are required for decreases in respiratory frequency that follow brief hypoxia. Changes in synchrony and firing rates of ventrolateral and midline medullary neurons may contribute to the long-term facilitation of breathing after brief intermittent hypoxia. Long-term changes in central respiratory motor control may occur after spinal cord injury, and the brain stem network implicated in the production of the respiratory rhythm could be reconfigured to produce the cough motor pattern. Preliminary analysis suggests that elements of brain stem respiratory neural networks respond differently to hypoxia and hypercapnia and interact with areas involved in cardiovascular control. Plasticity or alterations in these networks may contribute to the chronic upregulation of sympathetic nerve activity and hypertension in sleep apnea syndrome and may also be involved in sudden infant death syndrome. PMID- 12571147 TI - Selected contribution: Acute and sustained ventilatory responses to hypoxia in high-altitude natives living at sea level. AB - High-altitude (HA) natives have blunted ventilatory responses to hypoxia (HVR), but studies differ as to whether this blunting is lost when HA natives migrate to live at sea level (SL), possibly because HVR has been assessed with different durations of hypoxic exposure (acute vs. sustained). To investigate this, 50 HA natives (>3,500 m, for >20 yr) now resident at SL were compared with 50 SL natives as controls. Isocapnic HVR was assessed by using two protocols: protocol 1, progressive stepwise induction of hypoxia over 5-6 min; and protocol 2, sustained (20-min) hypoxia (end-tidal Po(2) = 50 Torr). Acute HVR was assessed from both protocols, and sustained HVR from protocol 2. For HA natives, acute HVR was 79% [95% confidence interval (CI): 52-106%, P = not significant] of SL controls for protocol 1 and 74% (95% CI: 52-96%, P < 0.05) for protocol 2. By contrast, sustained HVR after 20-min hypoxia was only 30% (95% CI: -7-67%, P < 0.001) of SL control values. The persistent blunting of HVR of HA natives resident at SL is substantially less to acute than to sustained hypoxia, when hypoxic ventilatory depression can develop. PMID- 12571148 TI - Selected contribution: High-altitude natives living at sea level acclimatize to high altitude like sea-level natives. AB - Sea-level (SL) natives acclimatizing to high altitude (HA) increase their acute ventilatory response to hypoxia (AHVR), but HA natives have values for AHVR below those for SL natives at SL (blunting). HA natives who live at SL retain some blunting of AHVR and have more marked blunting to sustained (20-min) hypoxia. This study addressed the question of what happens when HA natives resident at SL return to HA: do they acclimatize like SL natives or revert to the characteristics of HA natives? Fifteen HA natives resident at SL were studied, together with 15 SL natives as controls. Air-breathing end-tidal Pco(2) and AHVR were determined at SL. Subjects were then transported to 4,300 m, where these measurements were repeated on each of the following 5 days. There were no significant differences in the magnitude or time course of the changes in end tidal Pco(2) and AHVR between the two groups. We conclude that HA natives normally resident at SL undergo ventilatory acclimatization to HA in the same manner as SL natives. PMID- 12571149 TI - Selected contribution: Peripheral chemoreflex function in high-altitude natives and patients with chronic mountain sickness. AB - Peripheral chemoreflex function was studied in high-altitude (HA) natives at HA, in patients with chronic mountain sickness (CMS) at HA, and in sea-level (SL) natives at SL. Results were as follows. 1) Acute ventilatory responses to hypoxia (AHVR) in the HA and CMS groups were approximately one-third of those of the SL group. 2) In CMS patients, some indexes of AHVR were modestly, but significantly, lower than in healthy HA natives. 3) Prior oxygenation increased AHVR in all subject groups. 4) Neither low-dose dopamine nor somatostatin suppressed any component of ventilation that could not be suppressed by acute hyperoxia. 5) In all subject groups, the ventilatory response to hyperoxia was biphasic. Initially, ventilation fell but subsequently rose so that, by 20 min, ventilation was higher in hyperoxia than hypoxia for both HA and CMS subjects. 6) Peripheral chemoreflex stimulation of ventilation was modestly greater in HA and CMS subjects at an end-tidal Po(2) = 52.5 Torr than in SL natives at an end-tidal Po(2) = 100 Torr. 7) For the HA and CMS subjects combined, there was a strong correlation between end-tidal Pco(2) and hematocrit, which persisted after controlling for AHVR. PMID- 12571150 TI - Selected contribution: Ventilatory response to CO2 in high-altitude natives and patients with chronic mountain sickness. AB - The ventilatory responses to CO(2) of high-altitude (HA) natives and patients with chronic mountain sickness (CMS) were studied and compared with sea-level (SL) natives living at SL. A multifrequency binary sequence (MFBS) in end-tidal Pco(2) was employed to separate the fast (peripheral) and slow (central) components of the chemoreflex response. MFBS was imposed against a background of both euoxia (end-tidal Po(2) of 100 Torr) and hypoxia (52.5 Torr). Both total and central chemoreflex sensitivity to CO(2) in euoxia were higher in HA and CMS subjects compared with SL subjects. Peripheral chemoreflex sensitivity to CO(2) in euoxia was higher in HA subjects than in SL subjects. Hypoxia induced a greater increase in total chemoreflex sensitivity to CO(2) in SL subjects than in HA and CMS subjects, but peripheral chemoreflex sensitivity to CO(2) in hypoxia was no greater in SL subjects than in HA and CMS subjects. Values for the slow (central) time constant were significantly greater for HA and CMS subjects than for SL subjects. PMID- 12571151 TI - Plasma protein osmotic pressure equations for humans. PMID- 12571152 TI - Lung surfactants: in vitro vs. in vivo. PMID- 12571153 TI - Measuring lung liquid volume and secretion using radioiodinated serum albumin and blue dextran. PMID- 12571154 TI - Safety issues in assisted reproduction technology: Should men undergoing ICSI be screened for chromosome abnormalities in their sperm? AB - The incidence of aneuploidy in gametes of men undergoing ICSI has raised the prospect of there being risks associated with ICSI and the question of whether or not to screen men for sperm aneuploidy before treatment. We report results of a questionnaire undertaken to address how IVF staff perceive this problem, whether ICSI men are already being screened for sperm aneuploidy and the extent to which IVF specialists feel that there is merit in such a test. The results suggest that this is seen as a problem but most feel the risks outweigh the benefits. Most claimed their clinics do not screen sperm for aneuploidy but feel that there is merit in doing so. There are considerable benefits to screening i.e. couples would get additional information about the genetic repercussions of ICSI and could make informed decisions before treatment; screening would also facilitate the design of a large research study to give clearer answers on the safety of ICSI. However, we acknowledge counter arguments i.e. families would not necessarily benefit as most would have the ICSI procedure regardless of screen results; sex chromosome trisomies clinically are not severe enough to worry about in this context and there are other potential risks of ICSI that screening would not address. PMID- 12571155 TI - Postzygotic diploidization of triploids as a source of unusual cases of mosaicism, chimerism and twinning. AB - Triploidy is one of the most frequent chromosomal errors responsible for reproduction failure. This paper encompasses, in one conceptual frame, four recent findings in reproduction biology: predominant dispermic origin of triploids, paternal centrosome inheritance, eccentric cleavage divisions of dispermic triploid zygotes and certain intricate cases of mosaicism/chimerism. It is argued that dispermic zygotes, in contrast to digynic ones, are characterized by cytogenetic phenomenon described here as postzygotic diploidization of triploids (PDT). PDT embraces three main developmental scenarios: (i) the maintenance of the triploid state accompanied by regular segregation of 2n cells and the 2n/3n mixoploid populations; (ii) immediate diploidization with elimination of an odd haploid set of chromosomes and regular appearance of 1n/2n, 2n/3n and other mixoploids and (iii) tripolar spindle formation leading to gross aneuploidy, cell death with occasional survival of 2n+1 or 2n+1+1 trisomics and uniparental disomics. According to the PDT concept, a trisomy and disomy might occur due to generalized karyotype instability of dispermic triploids. PDT may provide a natural explanation for the regular appearance of 2n homozygous androgenic moles, various 2n/3n, 2n/2n molar/twin complexes without necessitating the concept of the 'empty' oocyte fertilization. Convincing evidence for a reservoir of anuclear oocytes does not exist. Peculiar implications are expected in the case of two rounds of diploidizations or involvement of triploid cell derivatives in the twinning process. Cryptic mosaic/chimeras and unusual twins intermediate between monozygotic (MZ) and dizygotic (DZ) are expected. Thus, PDT could have an explanation for the broad spectrum of odd reproductive cytogenetic events and might provide additional alternatives and definite predictions. PMID- 12571156 TI - Is genetic analysis useful in the routine management of hydatidiform mole? AB - Complete hydatidiform mole and partial hydatidiform mole are two abnormal conceptuses that may be identified by clinical, ultrasonographic, gross morphological, histological, and genetic characteristics. Among all these criteria, the specific diagnosis is generally confirmed only upon histological review. However, an accurate diagnosis based on morphological criteria is difficult and several studies have shown that misclassifications are frequent, even for experienced pathologists. An erroneous diagnosis may imply that women are either not enrolled in an adequate beta-hCG follow-up with the risk that hydatidiform mole (HM) progresses to choriocarcinoma, or are enrolled in an unnecessary follow-up. A reliable and complementary method to the pathologic interpretation is a genetic study of the conceptus to eliminate the diagnostic dilemma by distinguishing non-molar spontaneous abortions from HM and to define the type of HM. The aim of our study was to review the genetic basis of HM and discuss its relevance in the routine management of the disorder. PMID- 12571157 TI - A novel loss of function mutation in exon 10 of the FSH receptor gene causing hypergonadotrophic hypogonadism: clinical and molecular characteristics. AB - BACKGROUND: Inactivating mutations of the FSH receptor (FSHR) are a rare cause of hypergonadotrophic hypogonadism in women. Only one patient with primary amenorrhoea due to an FSHR gene mutation has been reported outside of Finland, where the prevalence of Ala189Val mutations is particularly high. METHODS AND RESULTS: Here, we describe the clinical, molecular genetic and functional characteristics associated with a novel inactivating mutation in exon 10 of the FSHR gene identified in a patient who presented with primary amenorrhoea at 17 years of age. The C to G transversion found at nucleotide 1043 causes a Pro348Arg substitution in the extracellular region of the FSHR and results in a mutant FSHR that is completely inactive in functional studies and that does not bind FSH. The proband exhibits apparent homozygosity for this recessive mutation. Her father is heterozygous for the mutation while analysis of exon 10 of the FSHR gene from her mother revealed only wild-type sequence. Chromosome painting was used to exclude deletions or rearrangements of 2p, and microsatellite markers did not show paternal uniparental isodisomy for this region. These findings suggest that the proband is hemizygous, with an inherited or de-novo microdeletion, or alternatively a de-novo gene conversion, of the accompanying FSHR allele. CONCLUSIONS: This case confirms the importance of the FSHR in female pubertal development and reproduction, and supports a relationship between phenotype and function for FSHR mutations. PMID- 12571158 TI - A simple, low cost and non-invasive method for screening Y-chromosome microdeletions in infertile men. AB - BACKGROUND: Recent investigations emphasized a high prevalence of Y-chromosome microdeletions in men having severely impaired spermatogenesis. Screening of these men is recommended prior to assisted reproduction techniques. METHODS: The aim of this study was to define a reliable and efficient method to detect Y chromosome deletions in infertile men. At first the feasibility of using a cytobrush to collect buccal cells as a source of DNA was tested. Then, a multiplex PCR in accordance with European recommendations (European Andrology Academia: EAA) was compared with a commercial kit. The test population consisted of 18 infertile male patients (with a known Y-deletion). Both buccal and blood cells were used for DNA extraction. A specific DNA extraction protocol was carried out on the buccal cells. RESULTS: Between 4-10 micro g of DNA were retrieved per brush, allowing for several PCR attempts. The commercial kit failed to detect an AZFa deletion. Furthermore, markers sY130, sY133 and sY153, included in the kit, are not reliable. Both false negative and false positive results were generated by the commercial kit. CONCLUSION: A multiplex PCR performed pursuant to EAA recommendations is proposed. When the testing is conducted with DNA extracted from buccal cells, this protocol is simple, accurate and affordable. PMID- 12571159 TI - Influence of the catechol-O-methyltransferase (COMT) codon 158 polymorphism on estrogen levels in women. AB - BACKGROUND: Catechol-O-methyltransferase (COMT) is the principal enzyme in the conjugation pathway for hydroxylated estrogens. We hypothesize that blood 17beta estradiol (E(2)) and estrone (E(1)) levels in postmenopausal women receiving an oral E(2) preparation are dependent on the enzyme activity of COMT. METHODS: To determine the influence of this enzyme on E(2) serum levels three groups of 12 selected from 159 healthy normotensive postmenopausal women were selected according to their codon 158 COMT genotype (COMT(HH), COMT(HL), COMT(LL)) which is known to be associated with enzyme activity. All selected women received one 2 mg tablet estradiol valerate and blood samples were taken before treatment and after 1, 3 and 48 h. RESULTS: After 3 h the serum levels of E(2) were significantly higher in women with the COMT(LL) genotype (median 69 pg/ml, range 58-91) and the COMT(HL) genotype (median 69 pg/ml, range 43-84) compared with women with the COMT(HH) genotype (median 45 pg/ml, range 15-68, P < 0.005). In a univariate analysis of variance, considering age, body weight, and COMT genotype, body weight (P = 0.034) and COMT genotype (P < 0.001) were independently related to the increase of serum E(2) levels, whereas age was not. CONCLUSIONS: Our data demonstrate that serum E(2) levels significantly correlate with the COMT genotype. Differences in COMT genotype might be involved in causing variable effects of estrogens on diseases such as hormone-dependent cancers, coronary heart disease and on efficacy of hormone replacement therapy. PMID- 12571160 TI - A polymorphism of the interleukin-6 gene promoter and idiopathic recurrent miscarriage. AB - BACKGROUND: Cytokines have been described to play a major role in the pathogenesis of idiopathic recurrent miscarriage (IRM). We investigated the association between IRM and a polymorphism of the interleukin-6 (IL-6) gene and IL-6 serum levels. METHODS: In a prospective case-control study, we studied 161 women with IRM and 124 healthy controls. Peripheral venous puncture, DNA extraction and PCR were employed to genotype women for the presence of a polymorphism at position -174 in the promoter region of IL-6. Serum IL-6 levels were assessed by a commercially available ELISA. RESULTS: Allele frequencies among women with IRM and controls were 63.4 and 58.1% respectively for allele G (wild type), and 36.6 and 41.9% respectively for allele C (mutant). No association between allele C and the occurrence of IRM was found (odds ratio 0.8; 95% confidence interval = 0.57-1.12; P = NS). IL-6 serum levels were not significantly different between genotypes and between the study and control groups. CONCLUSIONS: This is the first report on an IL-6 polymorphism in IRM. Although known to alter IL-6 expression, the IL-6 polymorphism investigated was not associated with IRM and alterations in IL-6 serum levels in a Middle-European Caucasian population. PMID- 12571161 TI - Reproductive genetic counselling in non-mosaic 47,XXY patients: implications for preimplantation or prenatal diagnosis: Case report and review. AB - With an incidence of approximately 1 in 500 male newborns, the 47,XXY genotype is one the most common sex chromosome anomalies. It is also the most frequent genetic cause of human infertility. Some non-mosaic 47,XXY patients have sperm production which allows infertility treatment to be offered by ICSI. Therefore, the risk of transmitting a chromosome anomaly to the next generation is an important problem in reproductive genetic counselling of these patients. Here, we report on a twin pregnancy where two karyotypically normal neonates 46,XX and 46,XY were born after the use of ICSI in assisted reproduction of a patient with a non-mosaic 47,XXY syndrome. To date, only 38 evolving pregnancies including the present cases, have been reported after ICSI using sperm from non-mosaic 47,XXY patients. Although these data are scarce, they suggest that the risk of chromosome anomaly in the offspring of these patients is low; hence, their reproductive genetic counselling can be reassuring, and management of the pregnancy can proceed with caution. PMID- 12571162 TI - Prenatal exposure to high galactose adversely affects initial gonadal pool of germ cells in rats. AB - BACKGROUND: In rats, prenatal exposure to high concentrations of galactose may contribute to a condition that is equivalent to the premature ovarian failure (POF) component of human galactosaemia. We investigated if development of POF under experimental galactosaemia-like conditions was attributed to impaired germ cell migration. METHODS: Pregnant rats were fed pellets supplemented with, or without, 35% galactose from day 3 of conception continuing through parturition. Between days 12-15, embryos from one uterine horn were dissected out. Primordial germ cells (PGC) were histochemically localized and counted on the basis of binding of Dolichos biflorus agglutinin, a lectin specific for terminal N acetylgalactosamine (GalNAc), to the surface glycoconjugate of the germ cells. The embryos from the other uterine horn were maintained until parturition. Liver activity of uridine diphosphate galactose 4-epimerase, the enzyme involved at multiple steps in the process of synthesis of GalNAc, was assayed in 1-2 day old female pups. RESULTS: The numbers of PGC at the day-specific sites on all days of examination were significantly lower (P /=11 mm was 4.2 +/- 0.3 in the rFSH group, 1.5 +/- 0.7 in the rLH group and 6.0 +/- 2.3 in the rFSH/rLH group (P = 0.07). 0/8 patients presented follicular growth arrest in the rFSH group, but 4/6 in the rLH group and 1/6 in the rFSH/rLH did. Study B: 5/12 patients presented follicular growth arrest in the rLH groups, but none in the placebo group. The mean number of follicles >/=11 mm was 4.6 +/- 1.8 for the placebo group, 2.5 +/- 1.9 for the rLH 225 IU group and 4.2 +/- 1.4 in the rLH 450 IU group (not significant). CONCLUSIONS: Results of this pilot study suggest that rLH alone can trigger follicular growth arrest in a significant number of patients, suggesting the existence of an 'LH ceiling' during late follicular maturation. PMID- 12571168 TI - Serum anti-Mullerian hormone is more strongly related to ovarian follicular status than serum inhibin B, estradiol, FSH and LH on day 3. AB - BACKGROUND: The study aim was to compare the relationship between serum anti Mullerian hormone (AMH) levels and other markers of ovarian function with early antral follicle count on day 3. METHODS: A total of 75 infertile women was studied prospectively. On cycle day 3, serum levels of AMH, inhibin B, estradiol (E(2)), FSH and LH levels were measured, and the number of early antral follicles (2-10 mm in diameter) estimated at ultrasound scanning to compare the strengths of hormonal-follicular correlations. RESULTS: Median (range) serum levels of AMH, inhibin B, E(2), FSH and LH were 1.39 ng/ml (0.24-6.40), 90 (16-182) pg/ml, 31 (15-111) pg/ml, 7.0 (2.9-19.3) mIU/ml and 4.7 (1.2-11.7) mIU/ml respectively, and follicular count was 12 (1-35). Serum AMH levels were more strongly correlated (P < 0.001) with follicular count (r = 0.74, P < 0.0001) than were serum levels of inhibin B (r = 0.29, P < 0.001), E(2) (r = -0.08, P = NS), FSH (r = -0.29, P < 0.001) and LH (r = 0.05, P = NS). CONCLUSIONS: Serum AMH levels were more robustly correlated with the number of early antral follicles than inhibin B, E(2), FSH and LH on cycle day 3. This suggests that AMH may reflect ovarian follicular status better than the usual hormone markers. PMID- 12571169 TI - Serum anti-Mullerian hormone dynamics during controlled ovarian hyperstimulation. AB - BACKGROUND: The study aim was to investigate possible changes in serum anti Mullerian hormone (AMH) levels during controlled ovarian hyperstimulation (COH), and their possible relationship with follicular development and other ovarian hormones. METHODS: A total of 93 women undergoing COH with GnRH agonist and FSH was studied prospectively. Serum levels of AMH, inhibin B, estradiol (E(2)), progesterone, testosterone and Delta(4)-androstenedione were measured when pituitary suppression was achieved (baseline), on days 6 and 8 of FSH treatment, and on the day of hCG. The number of small (<12 mm) and large (>/=12 mm) antral follicles were estimated using ultrasound. RESULTS: Serum AMH levels declined progressively (baseline, 1.21 +/- 0.11 ng/ml; day 6, 0.91 +/- 0.09 ng/ml; day 8, 0.77 +/- 0.08 ng/ml; and day of hCG, 0.53 +/- 0.06 ng/ml), whereas-as expected the other hormone levels increased during FSH treatment. Throughout COH, serum AMH levels correlated positively with the number of small but not large antral follicles, and with inhibin B serum levels. No correlation between AMH and the other hormones was observed. CONCLUSIONS: Serum AMH levels decline gradually during multiple follicular maturation, probably reflecting the dramatic reduction in the number of small antral follicles due to COH, and confirming the scarce AMH expression by larger follicles. PMID- 12571170 TI - Low 11-deoxycortisol to cortisol conversion reflects extra-adrenal factors in the majority of women with normo-gonadotrophic normo-estrogenic infertility. AB - BACKGROUND: Women with normogonadotrophic normo-estrogenic oligomenorrhoea often disclose a variety of clinical symptoms. Many of these individuals are obese with features of pseudo-hypercortisolism. In the current study, 11-deoxycortisol and cortisol concentrations were determined in this group and compared with ovulatory controls. METHODS AND RESULTS: Twenty-six women with clomiphene citrate-resistant infertility, 12 lean and 11 obese ovulatory controls were studied. Women with infertility had the highest 11-deoxycortisol concentrations (mean +/- SD: 4.1 +/- 1.5 ng/ml) compared with obese and lean controls (3.1 +/- 1.4 and 2.4 +/- 0.9 ng/ml) (P < 0.01), but similar morning cortisol concentrations (0.47 +/- 0.15, 0.45 +/- 0.16 and 0.47 +/- 0.18 nmol/l). Baseline 11-deoxycortisol/cortisol ratios (>90th percentile of ovulatory controls) were elevated in 23/26 infertile women (88%), and in 3/26 women (12%) after adrenocorticotrophic hormone (ACTH) stimulation. Three out of six lean infertile women had elevated baseline 11 deoxycortisol/cortisol ratios, but none of these women had elevated ratios after ACTH stimulation. Stepwise regression analysis, after exclusion of testosterone, revealed significant correlations between the groups (lean controls, obese controls, infertility) and ACTH-stimulated 11-deoxycortisol/cortisol ratio (P < 0.05), but not with fasting glucose, insulin, cortisol, 11-deoxycortisol and baseline 11-deoxycortisol/cortisol ratios. CONCLUSIONS: Congenital adrenal hyperplasia was not observed in the majority of infertile women. The data indicate that extra-adrenal factors were involved in most of the infertility syndromes that were studied. PMID- 12571171 TI - Acute elevation of plasma D-dimer levels associated with rupture of an ovarian endometriotic cyst: Case report. AB - A clinical case that suggested the involvement of blood coagulation cascades in the inflammation reaction induced by leakage of ovarian endometriotic cysts was encountered. Here, a rapid elevation in plasma D-dimer level (119.6 micro g/ml) was observed within 5 h after onset of ovarian cyst rupture. By contrast, the plasma fibrinogen level fell below normal range (105 mg/dl). During the following few days, the plasma D-dimer level fell acutely to approximately normal, while the fibrinogen level gradually increased to a high value (590 mg/dl). In addition, the anti-thrombin III level was slightly reduced on the day after rupture, though the concentration of blood platelets remained unchanged. Three days later, laparoscopic surgery showed the surface of the posterior wall of the uterus and peritoneum of the pelvic wall to be extensively covered with the leaked content of a right ovarian endometriotic cyst. Pathological examination confirmed the diagnosis of endometriotic cyst. In the endometrial stromal region, deposit of D-dimer was detected by immunohistochemical staining. These findings suggest that fibrin-derived degradation products in endometriotic cysts can affect coagulation cascades when they leak into the peritoneal cavity; they may also induce local inflammatory reactions, causing pelvic pain and/or extension of the endometriotic lesion. PMID- 12571172 TI - Developmental outcome at 2 years of age for children born after ICSI compared with children born after IVF. AB - BACKGROUND: Since the introduction of ICSI in 1991, medical outcome studies on ICSI children have been performed, but few have addressed developmental outcome. Hence, this outcome was assessed by performing a standard developmental test on children born after ICSI as compared with children born after IVF, at the age of 2 years. METHODS: In a prospective study, the medical and developmental outcome of 439 children born after ICSI (378 singletons, 61 twins) were compared with those of 207 children born after IVF (138 singletons, 69 twins), at the age of 24 28 months. These children were part of a cohort of children followed since birth. Of children reaching the age of 24-28 months between May 1995 and March 2002, 44.3% (2375/5356) were examined by a paediatrician who was unaware of the type of treatment used for each couple. Of all the children born, 12.2% (439/3618) in the ICSI group and 11.9% (207/1738) in the IVF group underwent a formal developmental assessment using the Bayley Scale of Infant Development (mental scale) by a paediatrician blinded to the type of treatment. RESULTS: There was no significant difference in maternal educational level, maternal age, gestational age, parity, birthweight, neonatal complication rate or malformation rate at 2 years between ICSI and IVF singletons, or between ICSI and IVF twins. No significant difference was observed in the developmental outcome using the Bayley scale at the age of 24 28 months (raw scores or test age) between ICSI children or IVF children. A multivariate regression analysis for the singleton children indicated that parity, sex (boys had lower scores than girls) and age had a significant influence on the test result, but that the fertility procedure (ICSI versus IVF) did not influence the test result. ICSI children from fathers with low sperm concentration, low sperm motility or poor morphology had a similar developmental outcome to that of children from fathers with normal sperm parameters. There were no significant differences between the initial cohort and the group lost to follow-up, nor between the psychologically tested and the non-tested group for a number of variables such as maternal educational level, birthweight in singletons and neonatal malformation rate. Although only some of the cohort of ICSI children were evaluated, a representative sample of both ICSI and IVF children was compared. CONCLUSIONS: There is no indication that ICSI children have a lower psychomotor development than IVF children. Paternal risk factors associated with male-factor infertility were found not to play a role in developmental outcome. PMID- 12571173 TI - Pregnancy course and health of children born after ICSI depending on parameters of male factor infertility. AB - BACKGROUND: There is an ongoing debate as to whether the outcome of ICSI is affected by the origin of sperm used or by the severity of male factor infertility. METHODS: Data from a multicentric prospective, controlled cohort study in Germany were analysed to look for differences in pregnancy course and outcome following the use of either ejaculated, epididymal or testicular sperm. Pregnancies were recruited before the 16th week of gestation and included only when they were still ongoing in the 16th week of gestation. A total of 2809 pregnancies was originally included in the study. For 2545 pregnancies with 3199 fetuses/children, data of sperm count and origin were available. RESULTS: Regarding pregnancy course and complications, there was no influence of the origin of sperm. No higher risk of pre-eclampsia was associated with the use of surgically obtained sperm. The birth data of children were similar between the three groups (ejaculated, epididymal, testicular sperm). The risk of major malformation in abortions, stillbirths, livebirths and induced abortions was not significantly different between the three groups, but these results must be treated with caution because of the low numbers in some groups. Major malformation rates did not depend on the number of sperm in the ejaculate. CONCLUSIONS: The course of pregnancy as well as the outcome after ICSI is neither affected by the origin of sperm nor by the number of sperm in the ejaculate. PMID- 12571174 TI - The potential value of tubal perfusion pressures measured during selective salpingography in predicting fertility. AB - BACKGROUND: The value of tubal perfusion pressures assessed during selective salpingography and tubal catheterization in predicting fertility has not been investigated. METHODS: A total of 325 infertile women underwent selective salpingography and tubal catheterization. Pregnancy information was collected in 256 (78.7%). The 50th (300 mmHg) and 90th (500 mmHg) centiles of the tubal perfusion pressure distribution in women with normal tubes on selective salpingography were used as thresholds. Women were divided into three tubal perfusion pressure groups: good (both tubes <300, or one tube <300 and the other 300-500 mmHg), mediocre (both tubes 300-500, or one tube <300 and the other >500 mmHg) and poor (both tubes >500, or one tube > 500 and the other 300-500 mmHg). RESULTS: The pregnancy rate in the good perfusion pressure group was significantly higher than that in the poor perfusion pressure group, both when all non-IVF/ICSI first conceptions (P = 0.001) as well as when spontaneous first conceptions only were considered (P = 0.010). The pregnancy rate in the mediocre group lay between the good and the poor groups, though none of the comparisons reached statistical significance. CONCLUSIONS: Selective salpingography can provide additional diagnostic information in comparison with other tubal assessment tests. Tubal perfusion pressures may be predictive of future fertility. PMID- 12571175 TI - Endometrial integrin expression in women undergoing IVF and ICSI: a comparison of the two groups and fertile controls. AB - BACKGROUND: Integrins are thought to play a vital role in implantation. Three integrins in particular (alpha(4)beta(1), alpha(v)beta(3) and alpha(1)beta(1)) are all present during the implantation window. Defects in their expression have been linked to tubal disease, unexplained infertility and endometriosis. Hence, a reduced endometrial integrin expression would be expected in women attending for IVF due to these causes of infertility when compared with those with male factor infertility attending for ICSI. METHODS: Women attending for IVF (n = 25) and ICSI (n = 25) treatment were recruited, and timed endometrial biopsies were taken during the 'implantation window' (cycle day 20-24). A group of fertile women (n = 15) attending for sterilization was used as controls. RESULTS: There was no significant difference in integrin expression between patients undergoing IVF or ICSI. Neither did these groups differ from the control group. CONCLUSIONS: The endometrium in patients undergoing ICSI treatment is sometimes thought to be more receptive, as the infertility might be due to a male factor. This study shows that there is no significant difference in integrin expression between patients attending for IVF or ICSI and the control group. These data add to the increasing uncertainty about the clinical value of assessing the endometrium with only one marker, in this case integrins. PMID- 12571176 TI - Pigtail catheter for the treatment of ascites associated with ovarian hyperstimulation syndrome. AB - BACKGROUND: Severe ovarian hyperstimulation syndrome (OHSS) is potentially dangerous. The study aim was to evaluate the efficacy and safety of percutaneous pigtail catheter drainage for the management of ascites complicating severe OHSS. METHODS: This was a prospective trial conducted at a private IVF centre and a tertiary teaching medical centre. A total of 26 patients with severe OHSS was recruited. Patients were divided into two groups. Patients in group 1 (n = 13) were hospitalized, while patients in group 2 (n = 13) were managed on an outpatient basis. A pigtail catheter was inserted under transabdominal ultrasound guidance and kept in place until drainage ceased. The main outcome measures were resolution of OHSS as determined by symptomatology and laboratory values, time to removal of catheter, patient tolerance of the procedure and complication rate. RESULTS: The catheter was successfully placed in all patients following one attempt and was kept in place for a mean +/- SD of 12.9 +/- 4.3 days (range 7 24). Average amount of fluid drained was 11.2 +/- 4.3 l (range: 3.35-18.5). An improvement of symptoms and signs was noted 24-48 h after catheter placement in all patients in both groups. Procedure was well tolerated and no complications reported. CONCLUSIONS: Percutaneous placement of a pigtail catheter is a safe and effective treatment modality for severe OHSS. It may represent an attractive alternative to multiple vaginal or abdominal paracentesis. PMID- 12571177 TI - Increased seminal plasma lead levels adversely affect the fertility potential of sperm in IVF. AB - BACKGROUND: Lead remains in high levels in the environment and is known to reduce fertility in animal models, but a direct link between lead exposures and human infertility has not yet been established. METHODS: In a prospective, double-blind study of the metal ion levels and sperm function, semen was obtained from partners of 140 consecutive women undergoing their first IVF cycle. Lead in seminal plasma was determined by atomic absorption spectroscopy. Motile sperm populations were assessed for surface receptors for mannose binding, and the ability to undergo premature ('spontaneous'), and free mannose-induced acrosome reactions. Fertile donor (n = 9) sperm were exposed to exogenous lead during capacitating incubations and then assessed for mannose receptor expression and acrosome loss. RESULTS: Lead levels were negatively correlated with IVF rates. Lead levels were negatively correlated to two of the three sperm function biomarkers (mannose receptors, mannose-induced acrosome reactions). Lead levels positively correlated with the spontaneous acrosome reaction. These findings were mimicked by in-vitro exposure of fertile donor sperm to lead. CONCLUSIONS: Multiple sperm parameters are affected as lead levels rise. Increased lead levels may contribute to the production of unexplained male infertility. PMID- 12571178 TI - Vitrification of human blastocysts using cryoloops: clinical outcome of 223 cycles. AB - BACKGROUND: The need to cryopreserve human blastocysts is increasing. The successful birth has been reported of a baby from a blastocyst vitrified using the cryoloop technique. The present study expands on this earlier report to confirm the effectiveness of this vitrification procedure. METHODS: In patients undergoing IVF at one of three clinics, supernumerary blastocysts on day 5 or 6 at various stages of development were vitrified using cryoloops. RESULTS: Of 725 vitrified blastocysts, 583 (80.4%) survived. After the transfer of 493 blastocysts in 207 cycles, 76 women (37%) became clinically pregnant. Among these women, 21 pregnancies ended in miscarriage, 23 healthy babies were born in 18 deliveries, and 37 pregnancies are ongoing. The survival rate of day 5 blastocysts (87%) was higher than that of day 6 blastocysts (55%), but implantation rates and pregnancy rates were not statistically significantly different. CONCLUSIONS: Clinical outcomes with 725 blastocysts and 207 transfers showed that vitrification using cryoloops is effective and practical for the cryopreservation of human blastocysts. Early blastocysts on day 5 seem to be the most suitable in terms of stage and age for cryopreservation, but developed and day 6 blastocysts can also be cryopreserved. PMID- 12571179 TI - Developmental potential of murine germinal vesicle stage cumulus-oocyte complexes following exposure to dimethylsulphoxide or cryopreservation: loss of membrane integrity of cumulus cells after thawing. AB - BACKGROUND: Cumulus cells of the cumulus-oocyte complex (COC) are important in oocyte maturation. Thus, in preserving immature oocytes it is prudent to also preserve their associated cumulus cells. The survival and function of oocytes and their associated cumulus cells was assessed following cryopreservation or exposure to cryoprotectant without freezing. METHODS: Immature COCs were collected from mice primed with pregnant mare's serum. COCs were either slow cooled or exposed to 1.5 mol/l dimethylsulphoxide without freezing. Treated and fresh COCs were stained for membrane integrity or, after in-vitro maturation and IVF, were assessed for developmental capability. Development of cumulus-denuded fresh oocytes, as well as denuded and frozen-thawed oocytes co-cultured with fresh cumulus cells, was assessed. RESULTS: Slow-cooled oocytes had significantly reduced coverage by intact cumulus cells compared with fresh COCs. Cumulus cell association and developmental capability were not substantially affected by exposure to cryoprotectant without freezing. Denuded fresh oocytes and cryopreserved COCs had decreased developmental potential that was not overcome by co-culture with fresh cumulus cells. CONCLUSIONS: Loss of association between oocyte and cumulus cells was induced by cryopreservation, but not by treatment with cryoprotectant alone. The data indicate that direct physical contact between cumulus cells and the oocyte, throughout maturation, improves subsequent embryo development. PMID- 12571180 TI - A morphological and chromosomal study of blastocysts developing from morphologically suboptimal human pre-embryos compared with control blastocysts. AB - BACKGROUND: IVF laboratories performing embryo transfer at day 2 or 3 after fertilization are currently discarding pre-embryos considered suboptimal using morphological criteria. The objective of this study was to investigate whether blastocysts, cultured from such pre-embryos (surplus), were chromosomally and morphologically normal. As a control group we used morphologically good quality embryos (GQE), cultured to the blastocyst stage. METHODS: Human pre-embryos considered suboptimal were cultured to the blastocyst stage. As a control group, frozen-thawed pre-embryos of good quality were cultured under identical conditions. The chromosomal status of the blastocysts obtained was studied by multi-colour fluorescence in-situ hybridization for chromosomes 13, 16, 18, 21, 22, X and Y. RESULTS: There is, on average, a significantly higher degree of chromosomal aberrations in blastocysts derived from surplus pre-embryos compared to blastocysts derived from GQE, and the chromosomal aberrations are generally found in a higher number of blastomeres per blastocyst. In addition, blastocysts from surplus pre-embryos had significantly poorer morphology compared to GQE. Improvement in morphology and/or developmental rate in surplus pre-embryos between day 2 and day 3 did not predict a morphologically/chromosomally normal blastocyst. However, this study shows that close to half of the surplus pre embryos that reach the blastocyst stage can be considered chromosomally normal when assessed for these seven chromosomes. Furthermore, we found that chromosomal aberrations were more concentrated in a particular cell population within blastocysts derived from GQE, compared with surplus blastocysts. CONCLUSIONS: The study suggests that even if the IVF laboratory is on average making the correct decision about the potential of a pre-embryo, surplus pre-embryos that might become chromosomally normal blastocysts are still being discarded. PMID- 12571181 TI - Effects of myo-inositol on the in-vitro maturation and subsequent development of mouse oocytes. AB - BACKGROUND: The study aim was to assess whether the incorporation of myo-inositol (MI) into culture medium could improve oocyte maturation in vitro. METHODS AND RESULTS: We performed a controlled prospective study using female ICR strain mice superovulated with pregnant mare's serum gonadotrophins. Cumulus-enclosed germinal vesicle (GV) oocytes were randomly cultured in medium with or without MI supplementation. The kinetics of GV breakdown after 4 h of incubation was significantly higher in oocytes incubated with 30 mmol/l of MI than in controls (P < 0.001). Accordingly, this concentration of MI was used for subsequent experiments. The proportion of metaphase II oocytes achieved after 24 h of culture, their fertilization and cleavage rates were significantly higher in the MI-treated group (P < 0.01, P < 0.05, P < 0.05 respectively). This group also demonstrated significant improvement in postimplantation development after transferring the 2-cell embryos to pseudopregnant mice. Confocal microscopy revealed spontaneous intracellular Ca(2+) oscillations within competent GV oocytes and treatment with MI caused an earlier onset of these Ca(2+) signals. CONCLUSIONS: Our results suggest that MI may affect meiotic progression of mouse GV oocytes possibly by enhancing the intracellular Ca(2+) oscillations. Supplementation of MI in culture medium may be useful for human oocyte maturation. PMID- 12571182 TI - Diminished ovarian reserve in female childhood cancer survivors with regular menstrual cycles and basal FSH <10 IU/l. AB - BACKGROUND: Approximately 70% of children treated for cancer are cured, however the treatment often impairs reproductive function. The aim of the present study was to assess ovarian reserve in women with an apparently normal ovarian function, who were cured of cancer in childhood. METHODS: Twenty-one female survivors with regular menstrual cycles and basal FSH <10 IU/l were included. The control group included 21 healthy age-matched regularly cycling women. On cycle day 2-5, ovarian volume and number of small antral follicles (2-5 mm) were evaluated with transvaginal ultrasonography. Repeated sonography, urinary LH testing, and endocrine assessments were performed during the actual cycle. Cycle length was calculated. RESULTS: The female survivors had significantly smaller volume per ovary (4.9 versus 6.8 cm(3); P < 0.01), a lower number of small antral follicles per ovary (4.5 versus 8.0; P < 0.01), and lower total number of follicles per ovary (8 versus 11, P < 0.01). Hormonal profiles were similar, but the mean cycle length of the female survivors was significantly shorter than in the control group (28.3 versus 31.0 days; P < 0.05). CONCLUSIONS: Childhood cancer survivors with regular menstrual cycles and basal FSH <10 IU/l seem to have a diminished ovarian reserve. Consequently, they may have a shortened reproductive span and an early menopause. PMID- 12571183 TI - Insulin-like growth factor (IGF)-1 and IGF binding protein-1 and -3 in the follicular fluid of infertile patients with endometriosis. AB - BACKGROUND: Endometriosis is associated with pituitary-ovarian axis dysfunction. The study of the follicular fluid in patients with endometriosis is important to elucidate the pathophysiological mechanism of this disease. The objective of this present paper was to analyse the dosages of insulin-like growth factor-1 (IGF-1) and IGF binding protein-1 and 3 (IGFBP-1 and IGFBP-3) in the follicular fluid environment of infertile patients with endometriosis. METHODS: A total of 41 infertile patients undergoing IVF between January 1999 and January 2000 participated in the cross-sectional prospective study. Patients were divided into three groups: group I, minimal/mild endometriosis (n = 12); group II, moderate/severe endometriosis (n = 10); and group III, tubal obstruction (n = 19). The ultra-short protocol was used in association with recombinant FSH for ovulation induction. Follicular fluid analysis was performed using radioimmunoassay with specific kits. RESULTS: Follicular fluid IGF-1 and IGFBP-3 levels were not significantly different among the groups; however, follicular fluid IGFBP-1 levels were lower in those patients with moderate/severe endometriosis (P < 0.05). Comparison of ovulation induction time, number of recombinant FSH units, number of follicles, oocytes and embryos, and fertilization and gestation/cycle rates showed non-significant differences. CONCLUSION: Infertile patients with moderate/severe endometriosis, which is associated with ovulatory dysfunction, presented lower levels of IGFBP-1 in the follicular fluid when undergoing IVF. PMID- 12571184 TI - Increased levels of interleukin-15 in the peritoneal fluid of women with endometriosis: inverse correlation with stage and depth of invasion. AB - BACKGROUND: Interleukin (IL)-15 is a novel cytokine with immunoregulatory and angiogenic properties. We compared IL-15 levels in the peritoneal fluid (PF) of women with and without endometriosis. METHODS: PF samples were obtained from 55 women with endometriosis (23 with superficial peritoneal implants, 19 with deep endometriotic implants and 13 with ovarian endometriomas). Eighteen women with normal pelvic anatomy undergoing tubal sterilization served as controls. RESULTS: PF IL-15 concentrations were increased in women with endometriosis (2.7 +/- 0.5 pg/ml) versus controls (2.1 +/- 0.3 pg/ml; P < 0.001). However, IL-15 levels were higher in women with superficial peritoneal implants (2.9 +/- 0.5 pg/ml) than women with deep endometriotic implants (2.6 +/- 0.4 pg/ml; P = 0.01) or ovarian endometriomas (2.2 +/- 0.4 pg/ml; P < 0.001). IL-15 was also higher in women with deep implants than in those with endometriomas (P < 0.05). PF IL-15 correlated inversely with both depth of invasion (r = -0.52) and the stage of endometriosis (r = -0.42). PF IL-15 levels demonstrated little variation during the menstrual cycle, and did not discriminate between women with infertility or pelvic pain. CONCLUSION: PF IL-15 levels are increased in women with endometriosis. However, IL-15 levels are inversely correlated with the depth of invasion and disease stage, suggesting a possible role for this cytokine in the early pathogenesis of endometriosis. PMID- 12571185 TI - Effect of various doses of gestogens on micronuclei frequency in human peripheral blood lymphocytes of pregnant women. AB - BACKGROUND: Since gestogens, in the form of hormonal substitution therapy, have been proposed to have a role in the prevention of threatened spontaneous abortions during the first three months of pregnancy, we decided to evaluate possible genotoxic effects of these preparations. METHODS: A total of 30 pregnant women, with a diagnosis of threatened spontaneous abortions, received the gestogen therapy in the first 3 months, and a sample of 30 pregnant women without indication for hormonal therapy were included as the control group. For investigation of mutagenic effects of gestogens in vivo the cytokinesis block (CB) micronucleus (MN) test was applied. RESULTS: Average MN frequency in the control group was 6.79 +/- 0.69 MN/1000 cells. The second analysed group included 12 patients with threatened spontaneous abortions, who received gestogen therapy in doses of 100-750 mg. Average MN frequency in these patients before therapy was 11.83 +/- 1.33 MN/1000 cells, and after therapy it was 16.50 +/- 1.32 MN/1000 cells (P < 0.001). The third analysed group comprised nine patients, who received gestogen therapy in doses of 750-2000 mg. Average MN frequency in these patients before therapy was 15.67 +/- 3.00 MN/1000 cells, and after therapy was 23.89 +/- 2.49 MN/1000 cells (P < 0.001). The fourth analysed sample comprised nine patients, treated with gestogen doses of 2000-8400 mg. The average MN frequency in these patients before therapy was 11.89 +/- 1.63 MN/1000 cells, and after therapy was 21.22 +/- 2.80 MN/1000 cells (P < 0.001). CONCLUSIONS: The increase of therapeutic gestogen doses was followed by an increase of average MN frequency. The greatest rise of MN frequency (1.8-fold) was observed in the group of patients who were treated with the highest gestogen doses (2000-8400 mg). The smallest increase (1.4-fold) of MN frequency was found in the group of patients whose therapeutic doses were the lowest (100-750 mg). PMID- 12571186 TI - Embryo reduction and birth weight discordance in dichorionic twins. AB - BACKGROUND: Twin birth weight discordance is associated with a poor perinatal outcome. The aim of this study was to analyse the risk factors of growth discordance among dichorionic twin pregnancies. METHODS: A cohort of 346 dichorionic twin pregnancies delivered at one perinatal centre between January 1996 and December 1999 was analysed. Two groups were created, according to the presence or absence of intra-pair birth weight discordance (n = 75 and 271 respectively). Birth weight discordance was defined as a difference of >/=20% of the weight of the heavier twin. The two groups were compared by uni- and multivariate analysis, with regard to the woman's characteristics, risk factors for growth restriction or discordance, and outcome of pregnancy. RESULTS: Pregnancies with birth weight discordance had a poor outcome compared with pregnancies without discordance, with a 4-fold increase in neonatal mortality. The rate of iatrogenic embryo reduction was significantly higher in discordant pregnancies (14.7 versus 6.6%, P = 0.03). The risk of birth weight discordance was increased with a larger starting number of embryos before reduction [20.2% (64/317), 28.6% (6/21), 57.1% (4/7) and 100% (1/1) respectively, for an initial number of two (no reduction), three, four, and five embryos, P = 0.02]. In multivariate analysis, embryo reduction was the only significant risk factor for the occurrence of birth weight discordance [adjusted odds ratio (OR) = 2.3 (1.0 5.2)]. CONCLUSIONS: Birth weight discordance carries a poor perinatal outcome. Embryo reduction is an independent risk factor for birth weight discordance in dichorionic twins. This finding emphasises the need for better control of assisted reproductive technology in order to avoid high-order multiple pregnancies. PMID- 12571187 TI - Quadruplet pregnancy following transfer of two embryos: Case report. AB - Following a long period of secondary unexplained infertility, a couple had assisted conception treatment with IVF and transfer of two embryos 2 days later. The woman conceived and was found to have a quadruplet pregnancy. Following spontaneous labour onset at 34 weeks gestation, she was delivered by immediate Caesarean section. All four babies (three boys, one girl) were eventually discharged home with the mother. All the boys were found to be of differing zygosity. The quadruplet pregnancy was attributed to natural conception following intercourse and fertilization of oocytes not collected at the time of IVF. The wisdom of intercourse during a cycle of IVF is discussed. PMID- 12571188 TI - Monozygotic twin delivery following reduction from quadramniotic-dichorionic gestation established after ICSI and embryo transfer: Case report. AB - We report the delivery of healthy monozygous (MZ) twins in a 31 year-old nulligravida following gonadotrophin ovulation induction, ICSI, assisted hatching and fresh embryo transfer. Although a sonogram on day 35 confirmed that two of four transferred embryos had implanted, a second transvaginal sonogram 1 week later showed each gestational sac had two conceptuses-all four were associated with distinct amniotic compartments. Cardiac activity was observed in all four embryos. At 12 weeks and 5 days gestation, chorionic villus sampling was performed on fetuses 1 and 2 which were euploid for chromosomes 13, 18, 21, X and Y via fluorescence in-situ hybridization analysis. Subsequent KCl injection into sacs 3 and 4 resulted in asystole for these fetuses, while cardiac activity in sacs 1 and 2 remained unchanged after reduction. A twin vaginal delivery occurred at 36 weeks gestation, resulting in the birth of two male infants and one placenta. This case represents the first known report of human quadruplet pregnancy consisting of two MZ twin sets conceived by assisted reproductive techniques. Our report reviews proposed mechanisms for explaining twinning, with special emphasis on zona pellucida micromanipulation and subsequent MZ twin induction. PMID- 12571189 TI - The association of age and semen quality in healthy men. AB - BACKGROUND: Although the effect of maternal age on fertility is well known, it is unclear whether paternal age also affects fertility. This cross-sectional study sought to characterize the association between age and semen quality, a well known proxy of fertility status. METHODS: A convenience sample of 97 non-smoking men (aged 22-80 years) without known fertility problems was recruited from a national government laboratory. The men provided semen samples and information relating to lifestyle, diet, medical and occupational details. Semen volume (ml), sperm concentration (x10(6)/ml), total sperm count (x10(6)), motility (%), progressive motility (%) and total progressively motile sperm count (x10(6)) were measured. RESULTS: After adjusting for covariates, semen volume decreased by 0.03 ml per year of age (95% CI: -0.05, -0.01); motility decreased by 0.7% per year (95% CI: -0.92, -0.43); progressive motility decreased by 3.1% per year (95% CI: 4.5, -1.6); and total progressively motile sperm count decreased by 4.7% per year (95% CI: -7.2, -2.2). There was a suggested decrease in sperm concentration and count. The proportion of men with abnormal volume, concentration and motility was significantly increased across the age decades. CONCLUSIONS: In a convenience sample of healthy men from a non-clinical setting, semen volume and sperm motility decreased continuously between 22-80 years of age, with no evidence of a threshold. PMID- 12571191 TI - Origin of sperm with extra chromosome set. PMID- 12571190 TI - Risks and complications in assisted reproduction techniques: Report of an ESHRE consensus meeting. AB - Assisted reproduction technique (ART) is an efficacious treatment in subfertile couples. So far little attention has been paid to the safety of ART, i.e. to its adverse events and complications. The consensus meeting on Risks and Complications in ART held in Maastricht in May 2002 focused on four topics: multiple pregnancies, long-term effects of ART on women, effects of ART on offspring, and morbidity/mortality registries. PMID- 12571193 TI - International standard for IVF centres. PMID- 12571194 TI - Sex selection by preimplantation genetic diagnosis: should it be carried out for social reasons? PMID- 12571198 TI - View of the ASRM Ethics Committee. PMID- 12571199 TI - Dobutamine-induced contractile reserve in stunned, hibernating, and scarred myocardium in patients with ischemic cardiomyopathy. AB - Because of damage to cardiomyocytes and the contractile apparatus, contractile reserve may be observed less frequently in hibernating than in stunned myocardium. The aim of this study was to assess the presence of contractile reserve in response to dobutamine infusion in a large group of patients with stunned and hibernating myocardium. METHODS: A total of 198 consecutive patients with ischemic cardiomyopathy (left ventricular ejection fraction < or = 40%) underwent resting 2-dimensional echocardiography to assess regional contractile dysfunction. On the basis of assessment of perfusion (with (99m)Tc-tetrofosmin SPECT) and glucose use (with (18)F-FDG SPECT), dysfunctional segments were grouped. Dysfunctional segments with normal perfusion were classified as stunned. Dysfunctional segments with a perfusion defect were classified as hibernating when a perfusion-(18)F-FDG mismatch was present. Dysfunctional segments with a perfusion defect were classified as scar tissue when a perfusion-(18)F-FDG match was present; these segments were subdivided into nontransmural and transmural scars. Contractile reserve was evaluated by dobutamine stress echocardiography. RESULTS: Dobutamine-induced contractile reserve was more frequently found in stunned than in hibernating myocardium (61% vs. 51%, respectively; P < 0.01). Only 14% of the scarred segments improved in wall motion during dobutamine infusion, significantly less than stunned or hibernating myocardium (P < 0.001). Nontransmural scars exhibited contractile reserve more frequently than did transmural scars. CONCLUSION: The progressive reduction of contractile reserve in stunned, hibernating, and scarred myocardium supports the hypothesis that stunning, hibernation, and scarring are not circumscript pathophysiologic entities but represent gradual ultrastructural damage on the myocyte level. PMID- 12571200 TI - Prognostic value of normal exercise and adenosine (99m)Tc-tetrofosmin SPECT imaging: results from the multicenter registry of 4,728 patients. AB - Event rates associated with a normal or low-risk myocardial perfusion SPECT imaging study have been shown by numerous investigators to be associated with <1%/y of follow-up. Thus, the aim of this study was to evaluate the prognostic value of a normal (99m)Tc-tetrofosmin study in a geographically diverse registry of patients undergoing stress myocardial perfusion SPECT. A total of 4,728 consecutively tested patients who underwent stress (99m)Tc-tetrofosmin SPECT at 5 U.S. hospitals were included in this series. METHODS: Patients were monitored for the occurrence of major cardiac events and hospitalizations. Uniform methods of data collection and standardized epidemiologic methods for follow-up were used at all centers. We used a risk-adjusted, Cox proportional hazards model to assess time to cardiac death. RESULTS: Of the 4,728 patients, one third underwent adenosine stress SPECT and two thirds underwent treadmill exercise. The observed annualized survival rate for those patients with a normal (99m)Tc-tetrofosmin study was 0.6%. Similarly, excellent survival rates were noted for the male and female subsets of this population as well as for patients who could exercise and for those undergoing pharmacologic stress testing. Compared with prior published outcomes studies on stress (201)Tl or (99m)Tc-sestamibi SPECT, the overall survival rates were similar and ranged from 99.3% to 99.7%. CONCLUSION: Results from this large multicenter registry provide further supportive evidence that the excellent prognosis associated with a normal SPECT scan is independent of the radiopharmaceutical used. PMID- 12571201 TI - Feasibility and image quality of dual-isotope SPECT using 18F-FDG and (99m)Tc tetrofosmin after acipimox administration. AB - Currently, with the rapidly increasing number of patients with heart failure due to chronic coronary artery disease, the need for viability studies to guide treatment in these patients is increasing. The most accurate method for viability assessment is metabolic imaging with (18)F-FDG with PET or SPECT. To obtain excellent image quality in all patients, the (18)F-FDG studies should be performed during hyperinsulinemic euglycemic clamping. However, this approach is time-consuming and is not feasible in busy nuclear medicine laboratories. Recently, the use of a nicotinic acid derivative, acipimox, has been suggested, but limited data are available on the image quality of the (18)F-FDG studies using this approach. METHODS: We evaluated the feasibility and image quality of (18)F-FDG SPECT (with dual-isotope simultaneous acquisition (DISA) using (99m)Tc tetrofosmin to assess perfusion) after acipimox administration in 50 nondiabetic patients. The image quality of both (18)F-FDG and (99m)Tc-tetrofosmin was assessed visually and quantitatively using myocardium-to-blood-pool (M/B) ratios as a measure of target-to-background ratio. The image quality and diagnostic value of DISA (99m)Tc-tetrofosmin SPECT was compared with standard (99m)Tc tetrofosmin SPECT at baseline. RESULTS: After acipimox administration, the plasma levels of free fatty acids were extremely low (68 +/- 89 nmol/L). No severe side effects were observed, only paroxysmal flushing. The (18)F-FDG image quality was good in 46 patients (92%) and moderate but still interpretable in the other 4 patients (8%). The clinical information of the baseline (99m)Tc-tetrofosmin SPECT was retained in the DISA (99m)Tc-tetrofosmin SPECT images because we did observe no substantial fill-in of perfusion defects by high (18)F-FDG uptake in the same segment. CONCLUSION: Cardiac (18)F-FDG SPECT after acipimox is safe and resulted consistently in good image quality; this simple approach may be the method of choice for routine cardiac metabolic imaging. PMID- 12571202 TI - Bicycle exercise stress in PET for assessment of coronary flow reserve: repeatability and comparison with adenosine stress. AB - PET allows absolute measurements of myocardial blood flow (MBF). The aim of the present study was to evaluate the feasibility and repeatability of supine bicycle exercise stress, compared with standard adenosine stress, in PET. METHODS: In 11 healthy volunteers, MBF was assessed at rest, during adenosine-induced (140 microg/kg/min over 7 min) hyperemia, and immediately after supine bicycle exercise (mean workload, 130 W, which is 70% of the predicted value) using PET and (15)O-H(2)O. The assessment was then repeated after 20 min. Coronary flow reserve (CFR) was calculated as hyperemic/resting MBF for adenosine stress and exercise stress. Repeatability was evaluated according to the method of Bland and Altman. RESULTS: No significant differences were found between the paired resting MBF (1.22 +/- 0.16 vs. 1.26 +/- 0.21 mL/min/g; mean difference, 3% +/- 11%) and the hyperemic MBF with adenosine stress (5.13 +/- 0.74 vs. 4.97 +/- 1.05; mean difference, -4% +/- 14%) or exercise stress (2.35 +/- 0.66 vs. 2.25 +/- 0.61; mean difference, -4% +/- 19%). CFR was reproducible with adenosine stress (4.23 +/- 0.62 vs. 4.05 +/- 1.06, P = not statistically significant; mean difference, 5% +/- 19%) and exercise stress (1.91 +/- 0.46 vs. 1.80 +/- 0.44, P = not statistically significant; mean difference, -5% +/- 15%). Repeatability coefficients for MBF were 0.26 (rest), 1.34 (adenosine stress), and 0.82 (exercise stress) mL/min/g. CONCLUSION: Assessment of CFR with (15)O-H(2)O and PET using bicycle exercise in the PET scanner is feasible and at least as repeatable as using adenosine stress. PMID- 12571203 TI - Performance of OSEM and depth-dependent resolution recovery algorithms for the evaluation of global left ventricular function in 201Tl gated myocardial perfusion SPECT. AB - It is unknown whether the use of ordered-subsets expectation maximization (OSEM) and depth-dependent resolution recovery (RR) will increase the accuracy of (201)Tl electrocardiogram-gated SPECT (GSPECT) for the measurement of global left ventricular (LV) function. METHODS: Fifty-six patients having both rest (201)Tl GSPECT and planar equilibrium radionuclide angiography (planar(RNA)) on the same day were studied. Twenty-nine patients also had LV conventional contrast angiography (Rx). LV ejection fraction (LVEF), end-diastolic volume (EDV), and end-systolic volume (ESV) were calculated with the quantitative gated SPECT software (QGS) using 4 different processing methods: filtered backprojection (FBP), OSEM, RR + FBP, and RR + OSEM. LVEF calculated with planar(RNA) and LV EDV and ESV calculated with Rx were considered gold standards. LVEF and volumes provided with the GSPECT methods were compared with the gold standard methods. RESULTS: LVEF calculated with GSPECT methods (FBP, OSEM, RR + FBP, and RR + OSEM) were similar (not statistically significant) and correlated well with planar(RNA). On Bland-Altman analysis, the mean +/- SD of absolute difference in LVEF with GSPECT FBP, OSEM, RR + FBP, and RR + OSEM methods versus planar(RNA) were similar, with relatively large limits of agreement. LV volumes calculated with the 4 GSPECT methods were significantly lower but correlated well with Rx LV volumes. LV volumes calculated with FBP and OSEM were lower than those calculated with RR + FBP and RR + OSEM (P < 0.01). On Bland-Altman analysis, the mean +/- SD of absolute difference in LV volumes with FBP, OSEM, RR + FBP, and RR + OSEM versus Rx was, respectively, 56 +/- 45 mL (P < 0.01 vs. the other 3 methods), 57 +/- 45 mL (P < 0.01 vs. the other 3 methods), 43 +/- 48 mL, and 46 +/- 47 mL, with correspondingly large limits of agreement. The variance of random error did not differ between FBP, OSEM, RR + FBP, and RR + OSEM for either LVEF or volumes. CONCLUSION: OSEM and FBP presented similar accuracy for LVEF and volume measured with the QGS software. Their combination with depth-dependent RR provided similar LVEF but more accurate LV volumes. PMID- 12571204 TI - Decreased 5-HT2a receptor binding in patients with anorexia nervosa. AB - Indirect estimations of brain neurotransmitters in patients with anorexia nervosa (AN) and low weight have demonstrated a reduction in brain serotonin (5-HT) turnover in general and led to hypotheses about dysfunction in the 5-HT(2a) receptor system. It was our aim to investigate the central 5-HT(2a) receptor binding index using SPECT brain imaging. METHODS: The 5-HT(2a) receptors of low weight patients with AN were studied by means of the highly specific radioiodinated 5-HT(2a) receptor antagonist 4-amino-N-[1-[3-(4 fluorophenoxy)propyl]-4-methyl-4-piperidinyl]-5-iodo-2-methoxybenzamide or (123)I 5-I-R91150. Fifteen patients with clinical diagnoses of AN and 11 age-matched healthy volunteers received intravenous injections of 185 MBq (123)I-5-I-R91150 and were scanned with high-resolution brain SPECT. RESULTS: Compared with healthy volunteers, patients with AN had a significantly reduced 5-HT(2a) binding index in the left frontal cortex, the left and right parietal cortex, and the left and right occipital cortex. A significant left-right asymmetry was noted in the frontal cortex (left < right). CONCLUSION: These results are in accordance with diminished metabolic and perfusion of frontal and parietal cortices reported in recent neuroimaging studies and imply localized disturbed serotonergic function. The data are discussed in the light of possible confounding factors related to the low-weight AN status. A regional cortical reduction in 5-HT(2a) binding index is not likely to be caused by a general reduction in serotonergic function due to the possible confounding factors. Suggestions for further research are given. PMID- 12571205 TI - Uptake in supraclavicular area fat ("USA-Fat"): description on 18F-FDG PET/CT. AB - The supraclavicular region is a common site for lymph node metastases. A commonly reported type of nonmalignant (18)F-FDG uptake on PET imaging in the supraclavicular region is "muscle uptake" purportedly due to muscle contraction in tense patients during the (18)F-FDG uptake phase. PET/CT offers the unique opportunity to correlate PET findings with CT anatomy in the supraclavicular region. METHODS: Images from the first 359 consecutive clinical whole-body studies (in 347 patients) using (18)F-FDG and a PET/CT scanner (with CT attenuation correction and ordered-subsets expectation maximization [OSEM] reconstruction) were retrospectively reviewed. The supraclavicular region was evaluated for the presence of abnormal uptake on PET images, and the corresponding CT findings were assessed. Three distinct patterns of abnormal (18)F-FDG uptake were noted: pattern A (uptake localizing to supraclavicular area fat [USA-fat], i.e., without corresponding lymph node or muscle uptake on CT), pattern B (uptake localizing to muscle on CT), and pattern C (uptake localizing to lymph nodes or soft-tissue masses on CT). RESULTS: Forty-nine patients (14.1%) (32 female, 17 male; mean age, 51.4 +/- 15.6 y; age range, 12-77 y) showed abnormal (18)F-FDG uptake in the supraclavicular region. Twenty patients (5.8%) had muscle uptake (group B); 15 (4.3%) had definite abnormal lymph nodes (group C). However, 14 patients (4.0%) had USA-fat (group A) and foci of very low Hounsfield units on CT. These foci were also present on (68)Ge attenuation corrected images (when obtained) and non-attenuation-corrected images. Uptake in USA-fat was typically bilateral and symmetric, intense, more often multifocal than linear, and located in fat on PET/CT. Age was not significantly different for group C versus the 2 other groups. Intensity; mean standardized uptake value, lean (SUV(L MEAN)); or maximum standardized uptake value, lean (SUV(L MAX)), did not allow differentiation between patterns A and C (P > 0.05). Standardized uptake values (SUV(L MAX), 3.1; SUV(L MEAN), 2.1) were significantly lower in group B than in the 2 other groups (P < 0.005). CONCLUSION: So-called muscle uptake in the supraclavicular region may be caused in a significant proportion of cases by an unrelated process we call the USA-fat finding, with (18)F-FDG uptake in tissues of low-Hounsfield (fat) density. This finding most likely reflects an underlying nonpathologic process that we hypothesize to be in foci of brown fat. This intense supraclavicular uptake should be recognized and should not be misinterpreted as a malignant metastatic process or as muscle uptake. PMID- 12571206 TI - 99mTc-aprotinin scintigraphy in amyloidosis. AB - Changes in the amount and distribution of amyloid lesions have been difficult to monitor because they can usually be demonstrated only by evident symptoms or from a biopsy. The recent progress in the treatment of amyloidosis stresses the need for an early diagnosis and the need for noninvasive monitoring during the course of treatment. To validate (99m)Tc-aprotinin scintigraphy, we studied 23 consecutive patients with known or suspected amyloidosis. METHODS: (99m)Tc Aprotinin (500-700 MBq) was injected intravenously and whole-body scans, regional images, and SPECT tomograms were obtained 90 min after tracer injection. RESULTS: Focal accumulations of (99m)Tc-aprotinin were seen in different organs of 22 patients with a total of 90 lesions, of which 20 were confirmed by biopsy or autopsy. Scintigraphy revealed "silent" amyloid deposits in at least 5 patients who later developed clinical symptoms. Physiologic uptake or excretion in liver and kidneys could not be differentiated from pathologic lesions in those organs. CONCLUSION: (99m)Tc-Aprotinin scintigraphy appears to be a fairly sensitive and specific diagnostic modality in patients with suspected amyloidosis. The technique is noninvasive, and it entails a minimal stress to the patient and is useful for detection of a wide range of lesions. PMID- 12571207 TI - Bone metastases in carcinoid tumors: clinical features, imaging characteristics, and markers of bone metabolism. AB - The purpose of this study was to describe the clinical presentation of bone metastases in patients with carcinoid tumors and to determine the diagnostic value of imaging techniques and markers of bone metabolism. METHODS: This retrospective study was performed on the entire group of patients with carcinoid tumors treated in our hospital from January 1992 to May 1999. Only patients with metastasized tumors were included. RESULTS: Eleven of 90 patients (12%) (95% confidence interval [CI], 5%-19%) with a metastasized carcinoid tumor had symptomatic bone metastases. All bone metastases occurred in 55 patients with midgut carcinoids (20%; 95% CI, 9%-31%). Plain radiography had a sensitivity of 44% (95% CI, 12%-76%); MRI, 100% (95% CI, 61%-100%); bone scintigraphy, 90% (95% CI, 72%-100%); and octreotide scintigraphy, 60% (95% CI, 35%-93%). In 9 patients, both octreotide scintigraphy and bone scintigraphy were performed. Of 45 bone lesions, 22 (49%) were visualized by both modalities, 13 (29%) were visualized with octreotide scintigraphy but not with bone scintigraphy, and 10 (22%) were visualized with bone scintigraphy but not with octreotide scintigraphy. In 2 patients, octreotide scintigraphy and bone scintigraphy provided complementary results. Markers of bone metabolism could not discriminate carcinoid patients from those without bone metastases. The markers of bone metabolism did not reflect the osteolytic or osteoblastic appearance of metastases. CONCLUSION: Pain is the principal symptom of bone metastases in patients with carcinoid tumors. Plain radiography and markers of bone metabolism do not contribute to the diagnosis of bone metastases. MRI has a high sensitivity for bone metastases. Both bone scintigraphy and octreotide scintigraphy have acceptable sensitivity and can provide complementary results. PMID- 12571208 TI - Improved prognostic value of 18F-FDG PET using a simple visual analysis of tumor characteristics in patients with cervical cancer. AB - In patients with cervical cancer, it is important to estimate prognosis at the time of diagnosis. This study using PET with (18)F-FDG was undertaken to determine whether a simple and fast visual analysis of characteristics of the primary tumor before initiation of treatment could achieve this goal. METHODS: Forty-seven patients with cervical cancer who were to be treated by combined radiation therapy and chemotherapy were imaged before beginning treatment. They were then followed for up to 3 y for evidence of recurrence or death. Images of the chest, abdomen, and pelvis were obtained 40-90 min after administration of 370-555 MBq (10-15 mCi) (18)F-FDG. Three observers then independently graded the primary tumor for size (0 = small, 1 = moderate, 2 = large), shape (0 = spherical, 1 = nonspherical), heterogeneity of uptake (0 = none, 1 = moderate, 2 = marked), and presence of lymph nodes (0 = none, 1 = pelvic, 2 = paraaortic, 3 = distant). The scores were summed to achieve a total score. A statistical calculation demonstrated that a score cutoff of 4 best separated patients with a good prognosis from patients with a bad prognosis. Kaplan-Meier analysis was used to compute progression-free survival and overall survival. Evaluation of lymph nodes alone was compared with the grading of tumor characteristics. RESULTS: Observers 1 and 2 scored 26 patients as having a good prognosis and 21 as having a bad prognosis. Observer 3 scored 30 and 17, respectively, a statistically insignificant difference. Survival curves were almost identical for the 3 observers. For progression-free survival, approximately 12% of patients with a good score had disease recurrence whereas approximately 75% with a bad score had disease recurrence. For overall survival, approximately 10% (good) and 80% (bad) died. Evaluation of lymph nodes also separated the groups, but not as well as did visual analysis alone. The combination of the 2 was only slightly superior to visual assessment alone. CONCLUSION: A simple, rapid, and highly reproducible system is described for visual grading of characteristics of the primary tumor in patients with cervical cancer at the time of diagnosis. This approach separates patients with a poor prognosis from those who will do well, thus providing a new tool for accurate estimation of prognosis. PMID- 12571209 TI - Prospective comparison of 18F-FDG PET with conventional imaging modalities (MRI, CT, and 67Ga scintigraphy) in assessment of combined intraarterial chemotherapy and radiotherapy for head and neck carcinoma. AB - To preserve the oral organs and functions in patients with head and neck carcinoma, accurate determination of the appropriate treatment after neoadjuvant chemotherapy and radiotherapy is of critical importance. We evaluated the diagnostic accuracy of (18)F-FDG PET relative to that of other conventional imaging modalities in the assessment of therapeutic response after combined intraarterial chemotherapy and radiotherapy as an organ preservation protocol. METHODS: The study was prospectively performed on 23 consecutive patients with head and neck squamous cell carcinoma who completed the treatment regimen and underwent 2 (18)F-FDG PET studies before and after neoadjuvant chemoradiotherapy. (67)Ga scintigraphy (only before therapy) as well as MRI and CT (both before and after therapy) were also performed. All images were blindly and independently interpreted without knowledge of histologic findings. The level of confidence in image interpretation was graded by means of a 5-point rating system (0 = definitely no tumor to 4 = definite tumor). RESULTS: Before treatment, (18)F-FDG PET detected primary tumors in all 23 patients and was more sensitive (100%) than MRI (18/23; 78.3%), CT (15/22; 68.2%), and (67)Ga scintigraphy (8/20; 40%), with a confidence level of 3 or 4 as a positive tumor finding. After chemoradiotherapy, residual tumors were histologically confirmed in 4 patients (pathologic complete response rate, 19/23; 82.6%). Although posttreatment (18)F FDG PET showed almost equal sensitivity (4/4; 100%) compared with MRI (3/3; 100%) or CT (3/4; 75%), its specificity (17/19; 89.5%) was superior to MRI (7/17, 41.2%) and to CT (10/17; 58.8%) for primary lesions. Regarding metastases to neck lymph nodes, only specificity for posttreatment images was calculated because no metastasis was confirmed in any patients after treatment. Six subjects had (18)F FDG PET-positive lymph nodes, which had pathologically no tumor cells and suggested an inflammatory reactive change after therapy. Therefore, the specificity of posttreatment (18)F-FDG PET (17/23; 73.9%) was almost identical to that of MRI (17/20; 85%) and CT (16/21; 76.2%) for neck metastasis. With combined chemoradiotherapy monitored with (18)F-FDG PET, 8 patients avoided surgery and the remaining 15 patients underwent a reduced form of surgery. CONCLUSION: (18)F FDG PET facilitates differentiation of residual tumors from treatment-related changes after chemoradiotherapy, which may be occasionally difficult to characterize by anatomic images. (18)F-FDG PET has a clinical impact for the management of patients with head and neck cancers after neoadjuvant chemoradiotherapy by optimizing surgical treatment for each patient and contributes to the improvement of the patient's quality of life. PMID- 12571210 TI - Percutaneous ethanol injection plus radioiodine versus radioiodine alone in the treatment of large toxic thyroid nodules. AB - Therapeutic options for toxic thyroid nodules (TTNs) are surgery, radioiodine (RAI), and percutaneous ethanol injection (PEI). Surgery is generally considered for TTNs larger than 4 cm. However, some patients may be at high surgical risk. The purpose of the study was to evaluate the efficacy of 2 nonsurgical modalities for these TTNs. METHODS: Twenty-two patients with TTNs larger than 4 cm were randomly assigned to 2 different treatments: to 11 (subgroup A), RAI was administered at a dose of 12,580 kBq/mL of nodular volume (NV) and was corrected for 100% 24-h (131)I uptake (RAIU); to 11 (subgroup B), 2-4 PEI sessions (ethanol injected = 30% NV) preceded 2 mo of 24-h RAIU and RAI dosing. Inclusion criteria were clinical and biochemical hyperthyroidism; a single palpable, hot nodule at (99m)Tc scintigraphy; and high surgical risk or refusal to have surgery. Patients gave informed consent. Local symptoms were evaluated by a previously validated score (symptom score, or SYS). RESULTS: Both treatments were well tolerated. Subgroup B showed a significant reduction of NV 2 mo after PEI: 33.6 +/- 18.5 versus 60.8 +/- 29.5 mL. Their 24-h RAIU was similar to that of subgroup A: 53.9 +/- 13.9 versus 61.8% +/- 11.0%. Consequently, the administered RAI dose was significantly lower for subgroup B (730 +/- 245 MBq) than for subgroup A (1,048 +/- 392 MBq). Twelve months after RAI, subgroup B had a higher NV reduction and a lower SYS than did subgroup A. In subgroup A, 1 patient was subclinically hyperthyroid, 2 showed a slight increase of thyroid-stimulating hormone, and 1 was clinically hypothyroid. In subgroup B, 1 patient had a slight increase of thyroid-stimulating hormone. CONCLUSION: We demonstrated that RAI, alone or with PEI, can be considered a valid alternative for TTNs larger than 4 cm when surgery is either refused or contraindicated. PEI plus RAI can be considered when marked shrinkage of a nodule is required or when reduction of the RAI dose can prevent hospitalization. PMID- 12571211 TI - Role of percutaneous ethanol injection in management of nodular lesions of the thyroid gland. PMID- 12571212 TI - 11C-acetate PET imaging in hepatocellular carcinoma and other liver masses. AB - It is well known that (18)F-FDG PET has a high average false-negative rate of 40% 50% in the detection of hepatocellular carcinoma (HCC). This is not an acceptable accuracy, particularly in countries where this tumor is prevalent. In this study, we evaluated prospectively the characteristics of (11)C-acetate and (18)F-FDG metabolism in HCC and other liver masses. METHODS: Fifty-seven patients were recruited into this study, with masses consisting of 39 HCC; 3 cholangiocarcinomas; 10 hepatic metastases from lung, breast, colon, and carcinoid primary malignancies; and 5 benign pathologies, including focal nodular hyperplasia (FNH), adenoma, and hemangioma. All patients, except 2 with typical findings of hemangioma and 3 clinically obvious metastases, were confirmed histopathologically by liver biopsy or resection. All patients fasted for at least 6 h and blood glucose concentration was measured before they underwent dual PET radiopharmaceutical evaluation of the upper abdomen with (11)C-acetate and (18)F-FDG. RESULTS: In the subgroup of HCC patients with the number of lesions < or = 3 (32 patients; 55 lesions; mean size +/- SD, 3.5 +/- 1.9 cm), the sensitivity of detection by (11)C-acetate is 87.3% ((11)C-acetate maximum SUV [SUV(max)] = 7.32 +/- 2.02, with a lesion-to-normal liver ratio of 1.96 +/- 0.63), whereas the sensitivity of detection by (18)F-FDG is only 47.3%, and 34% lesions show uptake of both tracers. None of the lesions was negative for both tracers (100% sensitivity using both tracers). In some lesions and in the subgroup of HCC patients (n = 7) with multifocal or diffuse disease, dual-tracer uptake by different parts of the tumor is demonstrated. Histopathologic correlation suggests that the well-differentiated HCC tumors are detected by (11)C-acetate and the poorly differentiated types are detected by (18)F-FDG. All 16 non-HCC malignant (cholangiocarcinoma and metastatic) liver lesions do not show abnormal (11)C-acetate metabolism. Of the benign liver lesions, only FNH shows mildly increased (11)C-acetate activities ((11)C-acetate SUV(max) = 3.59, with a lesion-to-normal liver ratio of 1.25). CONCLUSION: (11)C-Acetate has a high sensitivity and specificity as a radiotracer complementary to (18)F-FDG in PET imaging of HCC and evaluation of other liver masses. PMID- 12571213 TI - 11C-acetate: a new tracer for the evaluation of hepatocellular carcinoma. PMID- 12571214 TI - 18F-FDG PET evaluation of the response to therapy for lymphoma and for breast, lung, and colorectal carcinoma. AB - PET is a unique form of diagnostic imaging that observes in vivo biologic changes using radiopharmaceuticals that closely mimic endogenous molecules. (18)F-FDG, which allows the evaluation of glucose metabolism, is the most commonly used tracer in oncology because of the practical half-life of (18)F (110 min), compared with other short-lived positron emitters. (18)F-FDG uptake in tumors is proportional to the glycolytic metabolic rate of viable tumor cells indicating the increased metabolic demand of tumors for glucose. (18)F-FDG PET significantly improves the accuracy of imaging tumors in initial staging, management of recurrent cancer, and monitoring of therapy response. The information provided by this technique is more sensitive and specific than that provided by anatomic imaging modalities. (18)F-FDG PET is particularly superior to CT or MRI in the ability to evaluate the effectiveness of various treatment regimens early during therapy or after therapy. In this review, we discuss the role of (18)F-FDG PET in evaluating the response to therapy and the impact of this information on patient management. PMID- 12571215 TI - Clinically significant inaccurate localization of lesions with PET/CT: frequency in 300 patients. AB - This study evaluated lesion mislocalization between PET and CT on PET/CT studies when CT instead of germanium is used for attenuation correction (AC). METHODS: PET/CT scans were obtained for 300 clinical patients. Both CT and germanium scans were used to correct PET emission data. Cases were noted of suspected inaccurate localization of lesions on any of the 5 sets of images (PET using germanium AC [GeAC] fused and not fused with CT, PET using CT AC fused and not fused with CT, and PET with no AC [NAC]). Independent CT or MRI was used to determine true lesion locations. RESULTS: Six of 300 patients (2%) had lesion mislocalization when CT was used for AC or fusion. True liver dome lesions were mislocalized to the right lung base on PET/CT, likely because of a respiratory motion difference between PET and CT. No mislocalization was present on NAC PET or non-CT-fused GeAC PET images. CONCLUSION: Serious lesion mislocalization on PET/CT studies may occur, albeit very infrequently, when CT is used for either AC or fusion. PMID- 12571216 TI - Isoform selectivity of 3-125I-iodo-alpha-methyl-L-tyrosine membrane transport in human L-type amino acid transporters. AB - 3-(123)I-Iodo-alpha-methyl-L-tyrosine ((123)I-IMT) has been developed for SPECT of amino acid transport imaging. We examined the isoform selectivity of (125)I IMT transport of the 2 human L-type amino acid transporters, hLAT1 and hLAT2, with human 4F2hc-coexpressed Xenopus laevis oocytes. METHODS: An uptake study of (125)I-IMT was performed using transporter-expressed X. laevis oocytes. Oocytes were injected with 17.6 ng of hLAT1 or hLAT2 complementary RNA (cRNA) and 7.4 ng of h4F2hc cRNA in a molar ratio of 1:1. Two days after injection, the uptake of (125)I-IMT was measured in the Na(+)-free uptake solution containing 18.5 kBq of noncarrier-added (125)I-IMT. After incubation for 30 min at room temperature, radioactivity of the oocytes was determined. RESULTS: Of the 2 hLAT isoforms and h4F2hc-coexpressed X. laevis oocytes, (125)I-IMT uptake via hLAT1 was 5.95-fold higher than that via hLAT2 (P < 0.005). CONCLUSION: (125)I-IMT transport was hLAT1 selective. Investigations on the isoform selectivity of (125)I-IMT transport with other transporters are anticipated. PMID- 12571217 TI - Bone uptake studies in rabbits before and after high-dose treatment with 153Sm EDTMP or 186Re-HEDP. AB - The aim of this animal study was to measure the bone uptake of (99m)Tc hydroxymethylene diphosphonate (HDP) before and after high-dose treatment with (153)Sm-ethylenediaminetetramethylenephosphonate (EDTMP) or (186)Re-(tin)1,1 hydroxyethylidene diphosphonate (HEDP) to prove or disprove post-therapeutic alterations of bone uptake of radiolabeled bisphosphonates. METHODS: Quantitative bone scanning using 100 MBq (99m)Tc-HDP was performed on 12 rabbits before and 8 wk after radionuclide therapy with 1,000 MBq of either (153)Sm-EDTMP or (186)Re HEDP. Whole-body images were acquired at 3 min, 3 h, and 24 h after injection, and the activities for the whole body, urinary bladder, and soft tissue were measured by region-of-interest technique. From these data, bone uptake was calculated as initial whole-body activity minus urinary excretion and remainder soft-tissue activity. RESULTS: In animals treated with (153)Sm-EDTMP (n = 6), no differences could be proven for the bone uptake of (99m)Tc-HDP at 24 h after injection before and after therapy (51.1% +/- 5.5% vs. 48.0% +/- 6.1%, P > 0.05). There were also no significant differences for the remainder soft-tissue activities and the urinary excretion rates before and after therapy. Similar results were obtained in rabbits treated with (186)Re-HEDP: Bone uptake (44.8% +/ 6.7% vs. 40.4% +/- 4.9%, P > 0.05) and urinary excretion revealed no significant differences before and after treatment. CONCLUSION: No significant impairment of bone uptake of (99m)Tc-HDP could be observed 8 wk after high-dose radionuclide bone therapy. Because both the biokinetic data obtained for (186)Re-HEDP and (153)Sm-EDTMP and the myelotoxic effects were quite similar in rabbits to those in patients, it seems justifiable to expect the same result (i.e., no significant alteration of bone uptake of radiolabeled bisphosphonates) in patients undergoing a second radionuclide therapy within 2-3 mo after standard treatment with (186)Re HEDP or (153)Sm-EDTMP. PMID- 12571218 TI - Targeting of osseous sites with alpha-emitting 223Ra: comparison with the beta emitter 89Sr in mice. AB - The bone-seeking property and the potential exposure of red marrow by the alpha particle emitter (223)Ra (half-life, 11.43 d) were compared with those of the beta-emitter (89)Sr (half-life, 50.53 d). METHODS: The biodistributions of (223)Ra and (89)Sr were studied in mice. Tissue uptake was determined at 1 h, 6 h, 1 d, 3 d, and 14 d after intravenous administration. Radiation absorbed doses were calculated for soft tissues and for bone. Multicellular-level doses were estimated for bone marrow cavities. RESULTS: Both (89)Sr and (223)Ra selectively concentrated on bone surfaces relative to soft tissues. The measured bone uptake of (223)Ra was slightly higher than that of (89)Sr. At 24 h, the femur uptake of (223)Ra was 40.1% +/- 7.7% of the administered activity per gram of tissue. The uptake in spleen and most other soft tissues was higher for (223)Ra than for (89)Sr. Although predominant clearance of (223)Ra was observed from the soft tissues within the first 24 h, the bone uptake of (223)Ra, which was not significantly different from maximum after only 1 h, was not significantly reduced during the 14 d. Furthermore, little redistribution of (223)Ra daughter products away from bone was found (2% at 6 h and less than 1% at 3 d). Estimates of dose to marrow cavities showed that the (223)Ra alpha-emitter might have a marrow-sparing advantage compared with beta-emitters for targeting osteoid surfaces because the short-range alpha-particles irradiate a significantly lower fraction of the marrow volumes. At the same time, the bone surfaces will receive a therapeutically effective radiation dose. CONCLUSION: The results of this study indicate that (223)Ra is a promising candidate for high-linear-energy transfer alpha-particle irradiation of cancer cells on bone surfaces. (223)Ra can, together with its daughter radionuclides, deliver an intense and highly localized radiation dose to the bone surfaces with substantially less irradiation of healthy bone marrow compared with standard bone-seeking beta-emitters. PMID- 12571219 TI - Patient-specific, 3-dimensional dosimetry in non-Hodgkin's lymphoma patients treated with 131I-anti-B1 antibody: assessment of tumor dose-response. AB - A comprehensive, SPECT-based, patient-specific 3-dimensional (3D) dosimetry analysis has been performed using 3D-ID, a previously developed software package. The role of the total-body tumor burden, individual lesion size, tumor absorbed dose, and the spatial distribution of the absorbed dose on response and on the time course of tumor shrinkage has been examined in patients with lymphoma treated by radioimmunotherapy. METHODS: Data from 15 patients participating in a phase II study of (131)I-labeled anti-B1 antibody (tositumomab) were used. Patients were administered a tracer dose of (131)I for imaging and pharmacokinetics. Dose estimates from the tracer studies were used to prescribe the therapeutic administration such that the whole-body absorbed dose did not exceed 75 cGy. All patients received a fixed mass amount of antibody for both the tracer and the therapeutic administrations. SPECT and planar imaging were performed 3-4 d after the therapeutic administration. CT or MRI scans were available on all patients. Total tumor burden was assessed by drawing contours around all lymphoma lesions identified on whole-body CT or MRI. Mean absorbed doses were estimated for selected, index lesions by conventional dosimetry and also by 3D SPECT-based dosimetry. Using a patient-specific dosimetry package, 3D ID, dose-volume histograms were also generated to assess the spatial distribution of absorbed dose. This approach made it possible to obtain estimates of the minimum and maximum absorbed doses for individual tumors in addition to the mean. RESULTS: Mean absorbed dose estimates obtained by patient-specific SPECT-based dosimetry using 3D-ID were within 2%-5% of estimates obtained by conventional dosimetry. None of the absorbed dose parameters (mean, minimum, maximum, uniformity) were found to have a significant correlation with tumor response. The total-body tumor burden did not impact on overall response or toxicity. CONCLUSION: This analysis represents the first full reported implementation of a patient-specific 3D dosimetry package. The absence of a dose-response relationship for tumors is surprising and suggests that absorbed dose is not the sole determinant of tumor response in these patients. The absence of a correlation between the total-body tumor burden and overall response or toxicity suggests that tailoring the milligram amount of administered antibody to patient tumor burden is not likely to improve response or reduce toxicity. PMID- 12571220 TI - Evaluation of 18F-FA-4 and 11C-pipzA-4 as radioligands for the in vivo evaluation of the high-affinity choline uptake system. AB - 4,4'-Bis-1-hydroxy-2-(4-methylpiperidin-1-yl)ethyl-biphenyl (A-4), a tertiary amine analog of hemicholinium-3 (HC-3), is an inhibitor of the sodium-dependent high-affinity choline uptake (HACU) system. We have evaluated 4-[1-hydroxy-2-(4 (18)F-fluoromethylpiperidin-1-yl)ethyl]-4'-[1-hydroxy-2-(4-methylpiperidin-1 yl)ethyl]biphenyl ((18)F-FA-4) and 4-[1-hydroxy-2-(4-(11)C-methylpiperazin-1 yl)ethyl]-4'-[1-hydroxy-2-(4-methylpiperidin-1-yl)ethyl]biphenyl ((11)C-pipzA-4), an (18)F- and a (11)C-labeled derivative of A-4 as potential in vivo tracers for the HACU system. METHODS: The biodistribution of both compounds was determined in mice, and the intracerebral distribution was visualized by ex vivo and in vitro autoradiography. The in vitro affinity of the compounds was determined by a displacement study with (3)H-HC-3 on mice brain slices. RESULTS: In mice, both tracers show a high and persistent brain uptake. In vitro autoradiography shows binding to the striatum, whereas ex vivo autoradiography shows homogeneous binding throughout the brain. FA-4 and pipzA-4 inhibited the (3)H-HC-3 binding with a 50% inhibitory concentration of 57 nmol/L and 320 nmol/L, respectively. CONCLUSION: The evaluated compounds have affinity for HACU and show high uptake in the brain. In vitro binding of the compounds to the striatum cannot be inhibited by the presence of HC-3, whereas binding of HC-3 was inhibited by the presence of both FA-4 and pipzA-4, suggesting allosteric binding. PMID- 12571221 TI - Evaluating image reconstruction methods for tumor detection in 3-dimensional whole-body PET oncology imaging. AB - We compare 3 image reconstruction algorithms for use in 3-dimensional (3D) whole body PET oncology imaging. We have previously shown that combining Fourier rebinning (FORE) with 2-dimensional (2D) statistical image reconstruction via the ordered-subsets expectation-maximization (OSEM) and attenuation-weighted OSEM (AWOSEM) algorithms demonstrates improvements in image signal-to-noise ratios compared with the commonly used analytic 3D reprojection (3DRP) or FORE+FBP (2D filtered backprojection) reconstruction methods. To assess the impact of these reconstruction methods on detecting and localizing small lesions, we performed a human observer study comparing the different reconstruction methods. The observer study used the same volumetric visualization software tool that is used in clinical practice, instead of a planar viewing mode as is generally used with the standard receiver operating characteristic (ROC) methodology. This change in the human evaluation strategy disallowed the use of a ROC analysis, so instead we compared the fraction of actual targets found and reported (fraction-found) and also investigated the use of an alternative free-response operating characteristic (AFROC) analysis. METHODS: We used a non-Monte Carlo technique to generate 50 statistically accurate realizations of 3D whole-body PET data based on an extended mathematic cardiac torso (MCAT) phantom and with noise levels typical of clinical scans performed on a PET scanner. To each realization, we added 7 randomly located 1-cm-diameter lesions (targets) whose contrasts were varied to sample the range of detectability. These targets were inserted in 3 organs of interest: lungs, liver, and soft tissues. The images were reconstructed with 3 reconstruction strategies (FORE+OSEM, FORE+AWOSEM, and FORE+FBP). Five human observers reported (localized and rated) 7 targets within each volume image. An observer's performance accuracy with each algorithm was measured, as a function of the lesion contrast and organ type, by the fraction of those targets reported and by the area below the AFROC curve. This AFROC curve plots the fraction of reported targets at each rating threshold against the fraction of cases with (> or =1) similarly rated false reports. RESULTS: Images reconstructed with FORE+AWOSEM yielded the best overall target detection as compared with FORE+FBP and FORE+OSEM, although these differences in detectability were region specific. The FORE+FBP and FORE+AWOSEM algorithms had similar performances for liver targets. The FORE+OSEM algorithm performed significantly worse at target detection, especially in the liver. We speculate that this is the result of using an incorrect statistical model for OSEM and that the incorporation of attenuation weighting in AWOSEM largely compensates for this model inaccuracy. These results were consistent for both the fraction of actual targets found and the AFROC analysis. CONCLUSION: We demonstrated the efficacy of performing observer detection studies using the same visualization tools as those used in clinical PET oncology imaging. These studies demonstrated that the FORE+AWOSEM algorithm led to the best overall detection and localization performance for 1-cm-diameter targets compared with the FORE+OSEM and FORE+FBP algorithms. PMID- 12571222 TI - Determination of the attenuation map in emission tomography. AB - Reliable attenuation correction methods for quantitative emission CT (ECT) require accurate delineation of the body contour and often necessitate knowledge of internal anatomic structure. Two broad classes of methods have been used to calculate the attenuation map: transmission-less and transmission-based attenuation correction techniques. Whereas calculated attenuation correction belonging to the first class of methods is appropriate for brain studies, more adequate methods must be performed in clinical applications, where the attenuation coefficient distribution is not known a priori, and for areas of inhomogeneous attenuation such as the chest. Measured attenuation correction overcomes this problem and uses different approaches to determine this map, including transmission scanning, segmented magnetic resonance images, or appropriately scaled CT scans acquired either independently on separate or simultaneously on multimodality imaging systems. Combination of data acquired from different imagers suffers from the usual problems of working with multimodality images--namely, accurate co-registration from the different modalities and assignment of attenuation coefficients. A current trend in ECT is to use transmission scanning to reconstruct the attenuation map. Combined ECT/CT imaging is an interesting approach; however, it considerably complicates both the scanner design and the data acquisition and processing protocols. Moreover, the cost of such systems may be prohibitive for small nuclear medicine departments. A dramatic simplification could be made if the attenuation map could be obtained directly from the emission projections, without the use of a transmission scan. This is being investigated either using a statistical model of emission data or applying the consistency conditions that allow one to identify the operator of the problem and, thus, to reconstruct the attenuation map. This article presents the physical and methodologic basis of attenuation correction and summarizes recent developments in algorithms used to compute the attenuation map in ECT. Other potential applications are also discussed. PMID- 12571223 TI - Fractal analysis of nuclear medicine images again: validity and interpretation of results from new analysis methods. PMID- 12571224 TI - (99m)Tc-ciprofloxacin scintigraphy in rabbit model of prosthetic joint infection. PMID- 12571225 TI - Total-body and red marrow dose estimates. PMID- 12571226 TI - Mechanistic and mutational studies of Escherichia coli molybdopterin synthase clarify the final step of molybdopterin biosynthesis. AB - Biosynthesis of the molybdenum cofactor, a chelate of molybdenum or tungsten with a novel pterin, occurs in virtually all organisms including humans. In the cofactor, the metal is complexed to the unique cis-dithiolene moiety located on the pyran ring of molybdopterin. Escherichia coli molybdopterin synthase, the protein responsible for adding the dithiolene to a desulfo precursor termed precursor Z, is a dimer of dimers containing the MoaD and MoaE proteins. The sulfur used for dithiolene formation is carried in the form of a thiocarboxylate at the MoaD C terminus. Using an intein expression system for preparation of thiocarboxylated MoaD, the mechanism of the molybdopterin synthase reaction was examined. A stoichiometry of 2 molecules of thiocarboxylated MoaD per conversion of a single precursor Z molecule to molybdopterin was observed. Examination of several synthase variants bearing mutations in the MoaE subunit identified Lys 119 as a residue essential for activity and Arg-39 and Lys-126 as other residues critical for the reaction. An intermediate of the synthase reaction was identified and characterized. This intermediate remains tightly associated with the protein and is the predominant product formed by synthase containing the K126A variant of MoaE. Mass spectral data obtained from protein-bound intermediate are consistent with a monosulfurated structure that contains a terminal phosphate group similar to that present in molybdopterin. PMID- 12571227 TI - Structural studies of molybdopterin synthase provide insights into its catalytic mechanism. AB - Molybdenum cofactor biosynthesis is an evolutionarily conserved pathway present in eubacteria, archaea, and eukaryotes, including humans. Genetic deficiencies of enzymes involved in cofactor biosynthesis in humans lead to a severe and usually fatal disease. The molybdenum cofactor contains a tricyclic pyranopterin, termed molybdopterin, that bears the cis-dithiolene group responsible for molybdenum ligation. The dithiolene group of molybdopterin is generated by molybdopterin synthase, which consists of a large (MoaE) and small (MoaD) subunit. The crystal structure of molybdopterin synthase revealed a heterotetrameric enzyme in which the C terminus of each MoaD subunit is deeply inserted into a MoaE subunit to form the active site. In the activated form of the enzyme, the MoaD C terminus is present as a thiocarboxylate. The present study identified the position of the thiocarboxylate sulfur by exploiting the anomalous signal originating from the sulfur atom. The structure of molybdopterin synthase in a novel crystal form revealed a binding pocket for the terminal phosphate of molybdopterin, the product of the enzyme, and suggested a binding site for the pterin moiety present in precursor Z and molybdopterin. Finally, the crystal structure of the MoaE homodimer provides insights into the conformational changes accompanying binding of the MoaD subunit. PMID- 12571228 TI - Negative regulation of beta-catenin signaling by tyrosine phosphatase SHP-1 in intestinal epithelial cells. AB - Protein-tyrosine phosphatase SHP-1 is expressed at high levels in hematopoietic cells and at moderate levels in many other cell types including epithelial cells. Although SHP-1 has been shown to be a negative regulator of multiple signaling pathways in hematopoietic cells, very little is known about the biological role of SHP-1 in epithelial cells. In order to elucidate the mechanism(s) responsible for the loss of proliferative potential once committed intestinal epithelial cells begin to differentiate, the role and regulation of SHP-1 were analyzed in both intact epithelium as well as in well established intestinal cell models recapitulating the crypt-villus axis in vitro. Results show that SHP-1 was expressed in the nuclei of all intestinal epithelial cell models as well as in epithelial cells of intact human fetal jejunum and colon. Expression and phosphatase activity levels of SHP-1 were much more elevated in confluent growth arrested intestinal epithelial cells and in differentiated enterocytes as well. Overexpression of SHP-1 in intestinal epithelial crypt cells significantly inhibited dhfr, c-myc, and cyclin D1 gene expression but did not interfere with c fos gene expression. In contrast, a mutated inactive form of SHP-1 had no effect on these genes. SHP-1 expression significantly decreased beta-catenin/TCF dependent transcription in intestinal epithelial crypt cells. Immunoprecipitation experiments revealed that beta-catenin is one of the main binding partners and a substrate for SHP-1. Taken together, our results indicate that SHP-1 may be involved in the regulation of beta-catenin transcriptional function and in the negative control of intestinal epithelial cell proliferation. PMID- 12571229 TI - Bikunin target genes in ovarian cancer cells identified by microarray analysis. AB - Bikunin, a Kunitz-type protease inhibitor, could potentially suppress tumor cell invasion and metastasis. Our previous study revealed that overexpression of bikunin in a human ovarian cancer cell line, HRA, resulted in a down-regulation in uPA and uPAR gene expression. For identifying the full repertoire of bikunin regulated genes, a cDNA microarray hybridization screening was conducted using mRNA from bikunin-treated or bikunin-transfected HRA cells. A number of bikunin regulated genes were identified, and their regulation was confirmed by Northern blot analysis. Our screen identified 11 bikunin-stimulated genes and 29 bikunin repressed genes. The identified genes can indeed be classified into distinct subsets. These include transcriptional regulators, oncogenes/tumor suppressor genes, signaling molecules, growth/cell cycle, invasion/metastasis, cytokines, apoptosis, ion channels, extracellular matrix proteins, as well as some proteases. This screen identified suppression of several genes such as CDC-like kinase, LIM domain binding, Ets domain transcription factor, Rho GTPase activating protein, tyrosine phosphorylation-regulated kinase, hyaluronan-binding protein, matriptase, and pregnancy-associated plasma protein-A (PAPP-A), which have previously been implicated in enhancing tumor promotion. Northern blot analysis confirmed that several genes including matriptase and PAPP-A were down regulated by bikunin by approximately 9-fold. Further, genetic inhibition of matriptase or PAPP-A could lead to diminished invasion. These results show that bikunin alters the pattern of gene expression in HRA cells leading to a block in cell invasion. PMID- 12571230 TI - Helical structure of the needle of the type III secretion system of Shigella flexneri. AB - Gram-negative bacteria commonly interact with animal and plant hosts using type III secretion systems (TTSSs) for translocation of proteins into eukaryotic cells during infection. 10 of the 25 TTSS-encoding genes are homologous to components of the bacterial flagellar basal body, which the TTSS needle complex morphologically resembles. This indicates a common ancestry, although no TTSS sequence homologues for the genes encoding the flagellum are found. We here present an approximately 16-A structure of the central component, the needle, of the TTSS. Although the needle subunit is significantly smaller and shares no sequence homology with the flagellar hook and filament, it shares a common helical architecture ( approximately 5.6 subunits/turn, 24-A helical pitch). This common architecture implies that there will be further mechanistic analogies in the functioning of these two bacterial systems. PMID- 12571231 TI - The adhesion and differentiation-inhibitory activities of the immunoglobulin superfamily member, carcinoembryonic antigen, can be independently blocked. AB - The external domains of Ig superfamily members are involved in multiple binding interactions, both homophilic and heterophilic, that initiate molecular events leading to the execution of diverse cell functions. Human carcinoembryonic antigen (CEA), an Ig superfamily cell surface glycoprotein used widely as a clinical tumor marker, undergoes homophilic interactions that mediate intercellular adhesion. Recent evidence supports the view that deregulated overexpression of CEA has an instrumental role in tumorigenesis through the inhibition of cell differentiation and the disruption of tissue architecture. The CEA-mediated block of the myogenic differentiation of rat L6 myoblasts depends on homophilic binding of its external domains. We show here that L6 transfectant cells expressing CEA can "trans-block" the myogenesis of juxtaposed differentiation-competent L6 transfectant cells expressing a deletion mutant of CEA (DeltaNCEA). This result implies the efficacy of antiparallel CEA-CEA interactions between cells in the differentiation block. In addition, DeltaNCEA can acquire differentiation blocking activity by cross-linking with specific anti CEA antibodies, thus implying the efficacy of parallel CEA-CEA interactions on the same cell surface. The myogenic differentiation blocking activity of CEA was demonstrated by site-directed mutations to involve three subdomains of the amino terminal domain, shown previously to be critical for its intercellular adhesion function. Monovalent Fab fragments of monoclonal antibodies binding to the region bridging subdomains 1 and 2 could both inhibit intercellular adhesion and release the myogenic differentiation block. Amino acid substitutions Q80A, Q80R, and D82N in subdomain 3, QNDTG, however, were found to completely ablate the differentiation blocking activity of CEA but had no effect on intercellular adhesion activity. A cyclized peptide representing this subdomain was the most effective at releasing the differentiation block. PMID- 12571233 TI - Binding of zona binding inhibitory factor-1 (ZIF-1) from human follicular fluid on spermatozoa. AB - Previous studies showed that zona binding inhibitory factor-1 (ZIF-1) was the glycoprotein mainly responsible for the spermatozoa zona binding inhibitory activity of human follicular fluid. ZIF-1 has a number of properties similar to glycodelin-A. A binding kinetics experiment in the present study demonstrated the presence of two binding sites of ZIF-1 on human spermatozoa. These binding sites were saturable, reversible, and bound to (125)I-ZIF-1 in a time-, concentration-, and temperature-dependent manner. Glycodelin-A shared one common binding site with ZIF-1 on spermatozoa, and it could displace only 70% of the (125)I-ZIF-1 bound on human spermatozoa. ZIF-1 and glycodelin-A formed complexes with the soluble extract of human spermatozoa. Coincubation of solubilized zona pellucida proteins reduced the binding of ZIF-1 to two complexes of the extract, suggesting that the ZIF-1 binding sites and zona pellucida protein receptors on human spermatozoa were closely related. ZIF-1, but not glycodelin-A, significantly suppressed progesterone-induced acrosome reaction of human spermatozoa. The carbohydrate moieties derived from ZIF-1 reduced the binding of native ZIF-1 on human spermatozoa as well as the zona binding inhibitory activity of the glycoprotein, although the intensity of the effects are lower when compared with the native protein. These effects are not due to the action of the molecules on the motility, viability, and acrosomal status of the treated spermatozoa. Deglycosylated ZIF-1 had no inhibitory effect on both ZIF-1 binding and zona binding capacity of spermatozoa. We concluded that the carbohydrate part of ZIF-1 was critical for the functioning of the glycoprotein. PMID- 12571232 TI - A novel distal enhancer module regulated by pregnane X receptor/constitutive androstane receptor is essential for the maximal induction of CYP2B6 gene expression. AB - CYP2B6 plays an important role in the metabolism of a variety of structurally unrelated xenobiotics, including the anticancer drugs cyclophosphamide and ifosfamide. Previous studies have shown that the nuclear receptors constitutive androstane receptor (CAR) and pregnane X receptor (PXR) are involved in the transcriptional regulation of CYP2B genes through the phenobarbital-responsive enhancer module (PBREM). However, for human CYP2B6 the relatively weak response of the PBREM to PXR and CAR activation in transfection assays fails to describe the potent induction observed in primary human hepatocyte cultures. In this report, a novel nuclear receptor response module located -8.5 kilobases upstream from the CYP2B6 encoding region is described. Several potential PXR/CAR binding motifs were identified within the distal regulatory cluster. In electrophoretic mobility shift assays, one DR4 motif showed the strongest binding to both PXR and CAR. Transient transfection assays in HepG2 cells demonstrated that the novel distal response cluster could be activated by PXR and CAR. In primary human hepatocytes, both PBREM and the distal responsive element were activated individually by endogenous nuclear receptors upon exposure to prototypical inducers. However, in both HepG2 cells and primary human hepatocytes maximal reporter activation was observed in a construct containing both PBREM and the distal responsive element. In mouse tail-vein injection experiments, a construct containing both the distal responsive element and the proximal PBREM exhibited a strong synergistic expression in phenobarbital-treated mice. These results show that a novel xenobiotic-responsive enhancer module in the distal region of the CYP2B6 promoter (CYP2B6-XREM) together with the PBREM mediates optimal drug induced expression of CYP2B6. PMID- 12571234 TI - CXCR3 and heparin binding sites of the chemokine IP-10 (CXCL10). AB - The chemokine IP-10 (interferon-inducible protein of 10 kDa, CXCL10) binds the G protein-coupled receptor CXCR3, which is found mainly on activated T cells and NK cells, and plays an important role in Th1-type inflammatory diseases. IP-10 also binds to glycosaminoglycans (GAGs), an interaction thought to be important for its sequestration on endothelial and other cells. In this study, we performed an extensive mutational analysis to identify the CXCR3 and heparin binding sites of murine IP-10. The mutants were characterized for heparin binding, CXCR3 binding, and the ability to induce chemotaxis, Ca(2+) flux, and CXCR3 internalization. Double mutations neutralizing adjacent basic residues at the C terminus did not lead to a significant reduction in heparin binding, indicating that the main heparin binding site of IP-10 is not along the C-terminal alpha helix. Alanine exchange of Arg-22 had the largest effect on heparin binding, with residues Arg 20, Ile-24, Lys-26, Lys-46, and Lys-47 further contributing to heparin binding. A charge change mutation of Arg-22 resulted in further reduction in heparin binding. The N-terminal residue Arg-8, preceding the first cysteine, was critical for CXCR3 signaling. Mutations of charged and uncharged residues in the loop regions of residues 20-24 and 46-47, which caused reduced heparin binding, also resulted in reduced CXCR3 binding and signaling. CXCR3 expressing GAG-deficient Chinese hamster ovary cells revealed that GAG binding was not required for IP-10 binding and signaling through CXCR3, which suggests that the CXCR3 and heparin binding sites of IP-10 are partially overlapping. PMID- 12571235 TI - Purification, cloning, and characterization of XendoU, a novel endoribonuclease involved in processing of intron-encoded small nucleolar RNAs in Xenopus laevis. AB - Here we report the purification, from Xenopus laevis oocyte nuclear extracts, of a new endoribonuclease, XendoU, that is involved in the processing of the intron encoded box C/D U16 small nucleolar RNA (snoRNA) from its host pre-mRNA. Such an activity has never been reported before and has several uncommon features that make it quite a novel enzyme: it is poly(U)-specific, it requires Mn(2+) ions, and it produces molecules with 2'-3'-cyclic phosphate termini. Even if XendoU cleaves U-stretches, it displays some preferential cleavage on snoRNA precursor molecules. XendoU also participates in the biosynthesis of another intron-encoded snoRNA, U86, which is contained in the NOP56 gene of Xenopus laevis. A common feature of these snoRNAs is that their production is alternative to that of the mRNA, suggesting an important regulatory role for all the factors involved in the processing reaction. PMID- 12571236 TI - Phosphatidylinositol 3-kinases in carcinoembryonic antigen-related cellular adhesion molecule-mediated internalization of Neisseria gonorrhoeae. AB - Neisseria gonorrhoeae can be internalized by mammalian cells through interactions between bacterial opacity-associated (Opa) adhesins and members of the human carcinoembryonic antigen-related cellular adhesion molecule (CEACAM) family. We examined the role of phosphatidylinositol 3-kinases (PI3Ks) in gonococcal invasion of epithelial cell lines expressing either CEACAM1 or CEACAM3. CEACAM3 mediated internalization, but not that mediated by CEACAM1, was accompanied by localized and transient accumulation of the class I PI3K product phosphatidylinositol 3,4,5-trisphosphate at sites of bacterial engulfment. Inhibition of phosphatidylinositol 3-kinases reduced CEACAM3-mediated uptake but, paradoxically, led to an increase in intracellular survival of bacteria internalized via either CEACAM1 or CEACAM3, suggesting additional roles for PI3K products. Consistent with this finding, the class III PI3K product phosphatidylinositol 3-phosphate accumulated and persisted in the membrane of gonococcal phagosomes after internalization. Inhibition of PI3K blocked phagosomal acquisition of the late endosomal marker lysosome-associated membrane protein 2 and reduced phagosomal acidification. Inhibiting phagosomal acidification with concanamycin A also increased survival of intracellular gonococci. These results suggest two modes of action of phosphatidylinositol 3 kinases during internalization of gonococci: synthesis of phosphatidylinositol 3,4,5-trisphosphate is important for CEACAM3-mediated uptake, while phosphatidylinositol 3-phosphate is needed for phagosomal maturation and acidification, which are required for optimal bacterial killing. PMID- 12571237 TI - The quaternary structure of DNA polymerase epsilon from Saccharomyces cerevisiae. AB - DNA polymerase epsilon (Pol epsilon) from Saccharomyces cerevisiae consists of four subunits (Pol2, Dpb2, Dpb3, and Dpb4) and is essential for chromosomal DNA replication. Biochemical characterizations of Pol epsilon have been cumbersome due to protease sensitivity and the limited amounts of Pol epsilon in cells. We have developed a protocol for overexpression and purification of Pol epsilon from S. cerevisiae. The native four-subunit complex was purified to homogeneity by conventional chromatography. Pol epsilon was characterized biochemically by sedimentation velocity experiments and gel filtration experiments. The stoichiometry of the four subunits was estimated to be 1:1:1:1 from colloidal Coomassie-stained gels. Based on the sedimentation coefficient (11.9 S) and the Stokes radius (74.5 A), a molecular mass for Pol epsilon of 371 kDa was calculated, in good agreement with the calculated molecular mass of 379 kDa for a heterotetramer. Furthermore, analytical equilibrium ultracentrifugation experiments support the proposed heterotetrameric structure of Pol epsilon. Thus, both DNA polymerase delta and Pol epsilon are purified as monomeric complexes, in agreement with accumulating evidence that Pol delta and Pol epsilon are located on opposite strands of the eukaryotic replication fork. PMID- 12571238 TI - Calcium influx through receptor-operated channel induces mitochondria-triggered paraptotic cell death. AB - We address the specific role of cytoplasmic Ca(2+) overload as a cell death trigger by expressing a receptor-operated specific Ca(2+) channel, vanilloid receptor subtype 1 (VR1), in Jurkat cells. Ca(2+) uptake through the VR1 channel, but not capacitative Ca(2+) influx stimulated by the muscarinic type 1 receptor, induced sustained intracellular [Ca(2+)] rises, exposure of phosphatidylserine, and cell death. Ca(2+) influx was necessary and sufficient to induce mitochondrial damage, as assessed by opening of the permeability transition pore and collapse of the mitochondrial membrane potential. Ca(2+)-induced cell death was inhibited by ruthenium red, protonophore carbonyl cyanide m chlorophenylhydrazone, or cyclosporin A treatment, as well as by Bcl-2 expression, indicating that this process requires mitochondrial calcium uptake and permeability transition pore opening. Cell death occurred without caspase activation, oligonucleosomal/50-kilobase pair DNA cleavage, or release of cytochrome c or apoptosis inducer factor from mitochondria, but it required oxidative/nitrative stress. Thus, Ca(2+) influx triggers a distinct program of mitochondrial dysfunction leading to paraptotic cell death, which does not fulfill the criteria for either apoptosis or necrosis. PMID- 12571239 TI - 5-lipoxygenase-activating protein gene expression. Key role of CCAAT/enhancer binding proteins (C/EBP) in constitutive and tumor necrosis factor (TNF) alpha induced expression in THP-1 cells. AB - We examined expression of the 5-lipoxygenase activating protein (FLAP), which is critical for inflammatory cell leukotriene synthesis. A 3.4-kb segment of the FLAP gene 5'-untranslated region accounted for a 22-fold increase in promoter activity when transfected into the monocyte-like cell line, THP-1, and demonstrated no activity in non-inflammatory cells. Virtually all of the promoter activity was mediated by the first 134 bp upstream of the transcription start site, a region that contains CCAAT/enhancer-binding proteins (C/EBP) consensus binding sites, at -36 to -28 bp (distal) and -25 to -12 bp (proximal). DNase I footprint analyses demonstrated THP-1 nuclear extract proteins bind to the proximal site. Electrophoretic mobility shift assay analyses revealed that C/EBP alpha, delta, and epsilon bind to the proximal site and C/EBP alpha and epsilon bind to the distal site, constitutively. Transfection studies indicated that mutation of both the proximal and distal sites decreased constitutive FLAP promoter activity. Overexpression of C/EBP alpha, beta, and delta transactivated promoter activity and increased native FLAP mRNA accumulation. Mutation of both C/EBP sites essentially abolished promoter induction by C/EBP overexpression. Tumor necrosis factor (TNF) alpha induced FLAP mRNA expression, FLAP promoter activity, and C/EBP alpha, delta, and epsilon binding to the proximal and distal promoter consensus sites. Chromatin immunoprecipitation assays demonstrated that C/EBP alpha, delta, and epsilon bound to this region of the 5'-untranslated region, whereas C/EBP beta does not bind even under conditions of overexpression and stimulation. We conclude that the FLAP gene is transactivated by members of the C/EBP family of transcription factors in inflammatory cells and that these factors play an important role in FLAP gene induction by TNFalpha. PMID- 12571240 TI - Membrane fusion induced by vesicular stomatitis virus depends on histidine protonation. AB - Entry of enveloped animal viruses into their host cells always depends on a step of membrane fusion triggered by conformational changes in viral envelope glycoproteins. Vesicular stomatitis virus (VSV) infection is mediated by virus spike glycoprotein G, which induces membrane fusion at the acidic environment of the endosomal compartment. VSV-induced membrane fusion occurs at a very narrow pH range, between 6.2 and 5.8, suggesting that His protonation is required for this process. To investigate the role of His in VSV fusion, we chemically modified these residues using diethylpyrocarbonate (DEPC). We found that DEPC treatment inhibited membrane fusion mediated by VSV in a concentration-dependent manner and that the complete inhibition of fusion was fully reversed by incubation of modified virus with hydroxylamine. Fluorescence measurements showed that VSV modification with DEPC abolished pH-induced conformational changes in G protein, suggesting that His protonation drives G protein interaction with the target membrane at acidic pH. Mass spectrometry analysis of tryptic fragments of modified G protein allowed the identification of the putative active His residues. Using synthetic peptides, we showed that the modification of His-148 and His-149 by DEPC, as well as the substitution of these residues by Ala, completely inhibited peptide-induced fusion, suggesting the direct participation of these His in VSV fusion. PMID- 12571241 TI - Mot1 regulates the DNA binding activity of free TATA-binding protein in an ATP dependent manner. AB - Mot1 is an essential Snf2/Swi2-related Saccharomyces cerevisiae protein that binds the TATA-binding protein (TBP) and removes TBP from DNA using ATP hydrolysis. Mot1 functions in vivo both as a repressor and as an activator of transcription. Mot1 catalysis of TBP.DNA disruption is consistent with its function as a repressor, but the Mot1 mechanism of activation is unknown. To better understand the physiologic role of Mot1 and its enzymatic mechanism, MOT1 mutants were generated and tested for activity in vitro and in vivo. The results demonstrate a close correlation between the TBP.DNA disruption activity of Mot1 and its essential in vivo function. Previous results demonstrated a large overlap in the gene sets controlled by Mot1 and NC2. Mot1 and NC2 can co-occupy TBP.DNA in vitro, and NC2 binding does not impair Mot1-catalyzed disruption of the complex. Residues on the DNA-binding surface of TBP are important for Mot1 binding and the Mot1.TBP binary complex binds very poorly to DNA and does not dissociate in the presence of ATP. However, the binary complex binds DNA well in the presence of the transition state analog ADP-AlF(4). A model for Mot1 action is proposed in which ATP hydrolysis causes the Mot1 N terminus to displace the TATA box, leading to ejection of Mot1 and TBP from DNA. PMID- 12571242 TI - Evidence that cytochrome b559 mediates the oxidation of reduced plastoquinone in the dark. AB - The function of cytochrome b(559) in photosystem II (PSII) was investigated using a mutant created in tobacco in which the conserved phenylalanine at position 26 in the beta-subunit (PsbF) was changed to serine (Bock, R., Kossel, H., and Maliga, P. (1994) EMBO J. 13, 4623-4628). The mutant grew photoautotrophically, but the amount of PSII was reduced and the ultrastructure of the chloroplast was dramatically altered. Very few grana stacks were formed in the mutant. Although isolated PSII-enriched membrane fragments showed low PSII activity, electron paramagnetic resonance indicated the presence of functional PSII. Difference absorption spectra showed that the cytochrome b(559) contained heme. The plastoquinone pool was largely reduced in dark-adapted leaves of the mutant, based on chlorophyll fluorescence and thermoluminescence measurements. We therefore propose that cytochrome b(559) plays an important role in PSII by keeping the plastoquinone pool and thereby the acceptor side of PSII oxidized in the dark. Structural alterations as induced by the single Phe --> Ser point mutation in the transmembrane domain of PsbF evidently inhibit this function. PMID- 12571243 TI - A catalytic mechanism for D-Tyr-tRNATyr deacylase based on the crystal structure of Hemophilus influenzae HI0670. AB - D-Tyr-tRNA(Tyr) deacylase is an editing enzyme that removes d-tyrosine and other d-amino acids from charged tRNAs, thereby preventing incorrect incorporation of d amino acids into proteins. A model for the catalytic mechanism of this enzyme is proposed based on the crystal structure of the enzyme from Haemophilus influenzae determined at a 1.64-A resolution. Structural comparison of this dimeric enzyme with the very similar structure of the enzyme from Escherichia coli together with sequence analyses indicate that the active site is located in the dimer interface within a depression that includes an invariant threonine residue, Thr-80. The active site contains an oxyanion hole formed by the main chain nitrogen atoms of Thr-80 and Phe-79 and the side chain amide group of the invariant Gln-78. The Michaelis complex between the enzyme and D-Tyr-tRNA was modeled assuming a nucleophilic attack on the carbonyl carbon of D-Tyr by the Thr-80 O(gamma) atom and a role for the oxyanion hole in stabilizing the negatively charged tetrahedral transition states. The model is consistent with all of the available data on substrate specificity. Based on this model, we propose a substrate assisted acylation/deacylation-catalytic mechanism in which the amino group of the D-Tyr is deprotonated and serves as the general base. PMID- 12571244 TI - Protein kinase CK2 phosphorylates the high mobility group domain protein SSRP1, inducing the recognition of UV-damaged DNA. AB - The structure-specific recognition protein SSRP1 plays a role in transcription and replication in the chromatin context. Mediated by its C-terminal high mobility group (HMG) box domain, SSRP1 binds DNA non-sequence specifically but recognizes certain DNA structures. Using acetic acid urea polyacrylamide gel electrophoresis and mass spectrometry, we have examined the phosphorylation of maize SSRP1 by protein kinase CK2 alpha. The kinase phosphorylated several amino acid residues in the C-terminal part of the SSRP1 protein. Two phosphorylation sites were mapped in the very C-terminal region next to the HMG box domain, and about seven sites are localized within the acidic domain. Circular dichroism showed that the phosphorylation of the two C-terminal sites by CK2 alpha resulted in a structural change in the region of HMG box domain, because the negative peak of the CD spectrum at 222 nm was decreased by approximately 10%. In parallel, the phosphorylation induced the recognition of UV-damaged DNA, whereas the non phosphorylated protein does not discriminate between UV-damaged DNA and control DNA. The affinity of CK2 alpha-phosphorylated SSRP1 for the DNA correlates with the degree of UV-induced DNA damage. Moreover, maize SSRP1 can restore the increased UV-sensitivity of a yeast strain lacking the NHP6A/B HMG domain proteins to levels of the control strain. Collectively, these findings indicate a role for SSRP1 in the UV response of eukaryotic cells. PMID- 12571245 TI - Actin binding of human LIM and SH3 protein is regulated by cGMP- and cAMP dependent protein kinase phosphorylation on serine 146. AB - Various drugs that elevate cGMP levels and activate cGMP-dependent protein kinase (cGK) inhibit agonist-induced platelet activation. In the present study we identified the LIM and SH3 domain protein (LASP) that was recently cloned from human breast cancer cells (Tomasetto, C., Regnier, C., Moog-Lutz, C., Mattei, M. G., Chenard, M. P., Liderau, R., Basset, P., and Rio, M. C. (1995) Genomics 28, 367-376) as a novel substrate of cGK in human platelets. Recombinant human LASP was phosphorylated by cGMP- and cAMP-dependent protein kinase (cAK) in vitro. Cotransfection of PtK-2 cells with LASP and cGK confirmed phosphorylation of LASP in vivo. Studies with human LASP mutants identified serine 146 as a specific phosphorylation site for cGK and cAK in vivo. LASP is an actin-binding protein, and the phospho-LASP-mimicking mutant S146D showed reduced binding affinity for F actin in cosedimentation experiments. Immunofluorescence of transfected PtK2 cells demonstrated the localization of LASP in the tips of cell membrane extensions and at cell-cell contacts. Expression of the human LASP mutant S146D resulted in nearly complete relocalization to the cytosol and reduced migration of the cells. Taken together, these data suggest that phosphorylation of LASP by cGK and cAK may be involved in cytoskeletal organization and cell motility. PMID- 12571246 TI - ERK and RhoA differentially regulate pseudopodia growth and retraction during chemotaxis. AB - Nonmotile cells extend and retract pseudopodia-like structures in a random manner, whereas motile cells establish a single dominant pseudopodium in the direction of movement. This is a critical step necessary for cell migration and occurs prior to cell body translocation, yet little is known about how this process is regulated. Here we show that myosin II light chain (MLC) phosphorylation at its regulatory serine 19 is elevated in growing and retracting pseudopodia. MLC phosphorylation in the extending pseudopodium was associated with strong and persistent amplification of extracellular-regulated signal kinase (ERK) and MLC kinase activity, which specifically localized to the leading pseudopodium. Interestingly, inhibition of ERK or MLC kinase activity prevented MLC phosphorylation and pseudopodia extension but not retraction. In contrast, inhibition of RhoA activity specifically decreased pseudopodia retraction but not extension. Importantly, inhibition of RhoA activity specifically blocked MLC phosphorylation associated with retracting pseudopodia. Inhibition of either ERK or RhoA signals prevents chemotaxis, indicating that both pathways contribute to the establishment of cell polarity and migration. Together, these findings demonstrate that ERK and RhoA are distinct pathways that control pseudopodia extension and retraction, respectively, through differential modulation of MLC phosphorylation and contractile processes. PMID- 12571247 TI - Insight into the structure and function of the transferrin receptor from Neisseria meningitidis using microcalorimetric techniques. AB - The transferrin receptor of Neisseria meningitidis is composed of the transmembrane protein TbpA and the outer membrane protein TbpB. Both receptor proteins have the capacity to independently bind their ligand human transferrin (htf). To elucidate the specific role of these proteins in receptor function, isothermal titration calorimetry was used to study the interaction between purified TbpA, TbpB or the entire receptor (TbpA + TbpB) with holo- and apo-htf. The entire receptor was shown to contain a single high affinity htf-binding site on TbpA and approximately two lower affinity binding sites on TbpB. The binding sites appear to be independent. Purified TbpA was shown to have strong ligand preference for apo-htf, whereas TbpA in the receptor complex with TbpB preferentially binds the holo form of htf. The orientation of the ligand specificity of TbpA toward holo-htf is proposed to be the physiological function of TbpB. Furthermore, the thermodynamic mode of htf binding by TbpB of isotypes I and II was shown to be different. A protocol for the generation of active, histidine-tagged TbpB as well as its individual N- and C-terminal domains is presented. Both domains are shown to strongly interact with each other, and isothermal titration calorimetry and circular dichroism experiments provide clear evidence for this interaction causing conformational changes. The N-terminal domain of TbpB was shown to be the site of htf binding, whereas the C-terminal domain is not involved in binding. Furthermore, the interactions between TbpA and the different domains of TbpB have been demonstrated. PMID- 12571248 TI - YY1 transcription factor down-regulates expression of CCR5, a major coreceptor for HIV-1. AB - Expression of CCR5, a major coreceptor for human immunodeficiency virus type 1 (HIV-1), is regulated by a number of transcription factors. Here we report that the YY1 transcription factor down-regulates CCR5 promoter activity and that overexpression of YY1 reduces cell surface CCR5 expression and infectibility by R5-HIV-1. Because YY1 also down-regulates promoter activities of CXCR4, another major coreceptor for HIV-1 and HIV-1 long terminal repeat, this transcription factor may play a critical role in the pathogenesis of HIV-1 disease. PMID- 12571249 TI - Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. AB - Syndecan-4 is a transmembrane heparan sulfate proteoglycan that acts as a coreceptor with integrins in focal adhesion formation. The central region of syndecan-4 cytoplasmic domain (4V; LGKKPIYKK) binds phosphatidylinositol 4,5 bisphosphate, and together they regulate protein kinase C alpha (PKC alpha) activity. Syndecan 4V peptide directly potentiates PKC alpha activity, leading to "superactivation" of the enzyme, apparently through an interaction with its catalytic domain. We now have performed yeast two-hybrid and in vitro binding assays to determine the interaction sites between 4V and PKC alpha. Full-length PKC alpha weakly interacted with 4V by yeast two-hybrid assays, but PKC alpha constructs that lack the pseudosubstrate region or constructs of the whole catalytic domain interacted more strongly. A mutated 4V sequence (4V(YF): LGKKPIFKK) did not interact with PKC alpha, indicating that tyrosine 192 in the syndecan-4 cytoplasmic domain might be critical for this interaction. Further assays identified a novel interaction site in the C terminus of the catalytic domain of PKC alpha (amino acid sequence 513-672). This encompasses the autophosphorylation sites, which are implicated in activation and stability. Yeast two-hybrid data were confirmed by in vitro binding and coimmunoprecipitation assays. The interaction of syndecan-4 with PKC alpha appears unique since PKC delta and epsilon did not interact with 4V in yeast two hybrid assays or coimmunoprecipitate with syndecan-4. Finally, overexpression of syndecan-4 in rat embryo fibroblast cells, but not expression of the YF mutant, increased PKC alpha localization to focal adhesions. The data support a mechanism where syndecan-4 binds PKC alpha and localizes it to focal adhesions, whose assembly may be regulated by the kinase. PMID- 12571250 TI - Endogenous association of TRAF2, TRAF3, cIAP1, and Smac with lymphotoxin beta receptor reveals a novel mechanism of apoptosis. AB - Lymphotoxin-beta receptor (LT beta R) is a member of tumor necrosis factor receptor family and plays essential roles in the embryonic development and organization of secondary lymphoid tissues. It binds two types of tumor necrosis factor family cytokines, heterotrimer LT alpha 1 beta 2 and homotrimer LIGHT, and activates multiple signaling pathways including transcriptional factor NF kappa B, c-Jun N-terminal kinase, and cell death. However, the molecular mechanism of the activation of these signaling pathways by LT beta R is not clear. Because there is no enzymatic activity associated with the receptor itself, the signal transduction of LT beta R is mediated by cytoplasmic proteins recruited to receptors. To identify these proteins, we took a proteomic approach. The endogenous LIGHT.LT beta R complex was affinity-purified from U937 cells, and proteins associated with the complex were identified by mass spectrometry. Four of five proteins identified, TRAF2, TRAF3, cIAP1, and Smac, are reported here. Their association with LT beta R was further confirmed by coimmunoprecipitation in U937 cells and HEK293 cells. The presence of cIAP1 and Smac in LIGHT.LT beta R complex revealed a novel mechanism of LIGHT.LT beta R-induced apoptosis. PMID- 12571251 TI - Syndecan-1 and -4 synthesized simultaneously by mouse mammary gland epithelial cells bear heparan sulfate chains that are apparently structurally indistinguishable. AB - Many of the biological functions attributed to cell surface heparan sulfate (HS) proteoglycans, including the Syndecan family, are elicited through the interaction of their HS chains with soluble extracellular molecules. Tightly controlled, cell-specific sulfation and epimerization of HS precursors endows these chains with highly sulfated, iduronate-rich regions, which are major determinants of cytokine and matrix-protein binding and which are interspersed by N-acetylated, poorly sulfated regions. Until this study, there have been no comprehensive structural comparisons made on HS chains decorating simultaneously expressed, but different, syndecan core proteins. In this paper we demonstrate that the HS chains on affinity-purified syndecan-1 and -4 from murine mammary gland cells are essentially identical by a number of parameters. Size determination, disaccharide analyses, enzymatic and chemical scission methods, and affinity co-electrophoresis all failed to reveal any significant differences in fine structure, domain organization, or ligand-binding properties of these HS species. These findings lead us to suggest that the imposition of the fine structure onto HS occurs independently of the core protein to which it is attached and that these core proteins, in addition to the HS chains, may play a pivotal role in the various biological functions ascribed to these macromolecules. PMID- 12571252 TI - Fibroblast growth factor-2-induced signaling through lipid raft-associated fibroblast growth factor receptor substrate 2 (FRS2). AB - The plasma membrane is not homogeneous but contains specific subcompartments characterized by their unique lipid and protein composition. Based on their enrichment in various signaling molecules, these membrane microdomains are recognized to be sites of localized signal transduction for a number of extracellular stimuli. We have previously shown that fibroblast growth factor-2 (FGF2) induced a specific signaling response within a lipid raft membrane microdomain in human neuroblastoma cells characterized by the tyrosine phosphorylation of a p80 phosphoprotein. Herein, we show that this protein is the signaling adaptor FRS2 and that it is localized exclusively to lipid rafts in vitro and in vivo. We have examined how the tyrosine phosphorylation and serine threonine phosphorylation of FRS2 within lipid rafts affect the response of cells to FGF2 signaling. Our data suggest that activation of protein kinase C, Src family kinases, and MEK1/2 are involved in regulating serine-threonine phosphorylation of FRS2, which can indirectly affect FRS2 phosphotyrosine levels. We also show that Grb2 is recruited to lipid rafts during signaling events and that activation of MEK1/2 by different mechanisms within lipid rafts may lead to different cellular responses. This work suggests that compartmentalized signaling within lipid rafts may provide a level of specificity for growth factor signaling. PMID- 12571253 TI - Connective tissue growth factor gene regulation. Requirements for its induction by transforming growth factor-beta 2 in fibroblasts. AB - In skin, the profibrotic protein connective tissue growth factor (CTGF) is not normally expressed. However, when skin cells are exposed to transforming growth factor-beta (TGF-beta), CTGF is induced in fibroblasts but not in epithelial cells. We have begun to investigate the requirements for the fibroblast-selective induction of CTGF by TGF-beta. Previously we found that this response was Smad dependent. Now we show that protein kinase C and Ras/MEK/ERK are necessary for the TGF-beta induction of the CTGF promoter but not of a generic Smad-responsive promoter (SBE-lux). Induction of the CTGF promoter is antagonized by c-Jun or by MEKK1, suggesting that a proper balance between the Ras/MEK/ERK and JNK MAPK cascades is necessary for TGF-beta induction of CTGF. We identify the minimal CTGF promoter element necessary and sufficient to confer TGF-beta responsiveness to a heterologous promoter and show that a tandem repeat of a consensus transcription enhancer factor binding element, 5'-GAGGAATGG-3', is necessary for this induction. This element has not been previously shown to play a role in TGF beta induction of gene expression in fibroblasts. Gel shift analysis shows that this sequence binds nuclear factors that are greatly enriched in fibroblasts relative to epithelial cells. Thus Smads, Ras/MEK/ERK, protein kinase C, and fibroblast-enriched factors that bind GAGGAATGG act together to drive the TGF beta-mediated induction of CTGF in fibroblasts. PMID- 12571254 TI - Cocaine and the heart. PMID- 12571255 TI - Adding a test for human papillomavirus DNA to cervical-cancer screening. PMID- 12571256 TI - Nephrotoxic effects in high-risk patients undergoing angiography. AB - BACKGROUND: The use of iodinated contrast medium can result in nephropathy. Whether iso-osmolar contrast medium is less nephrotoxic than low-osmolar contrast medium in high-risk patients is uncertain. METHODS: We conducted a randomized, double-blind, prospective, multicenter study comparing the nephrotoxic effects of an iso-osmolar, dimeric, nonionic contrast medium, iodixanol, with those of a low osmolar, nonionic, monomeric contrast medium, iohexol. The study involved 129 patients with diabetes with serum creatinine concentrations of 1.5 to 3.5 mg per deciliter who underwent coronary or aortofemoral angiography. The primary end point was the peak increase from base line in the creatinine concentration during the three days after angiography. Other end points were an increase in the creatinine concentration of 0.5 mg per deciliter or more, an increase of 1.0 mg per deciliter or more, and a change in the creatinine concentration from day 0 to day 7. RESULTS: The creatinine concentration increased significantly less in patients who received iodixanol. From day 0 to day 3, the mean peak increase in creatinine was 0.13 mg per deciliter in the iodixanol group and 0.55 mg per deciliter in the iohexol group (P=0.001; the increase with iodixanol minus the increase with iohexol, -0.42 mg per deciliter [95 percent confidence interval, 0.73 to -0.22]). Two of the 64 patients in the iodixanol group (3 percent) had an increase in the creatinine concentration of 0.5 mg per deciliter or more, as compared with 17 of the 65 patients in the iohexol group (26 percent) (P=0.002; odds ratio for such an increase in the iodixanol group, 0.09 [95 percent confidence interval, 0.02 to 0.41]). No patient receiving iodixanol had an increase of 1.0 mg per deciliter or more, but 10 patients in the iohexol group (15 percent) did. The mean change in the creatinine concentration from day 0 to day 7 was 0.07 mg per deciliter in the iodixanol group and 0.24 mg per deciliter in the iohexol group (P=0.003; value in the iodixanol group minus the value in the iohexol group, -0.17 mg per deciliter [95 percent confidence interval, -0.34 to -0.07]). CONCLUSIONS: Nephropathy induced by contrast medium may be less likely to develop in high-risk patients when iodixanol is used rather than a low osmolar, nonionic contrast medium. PMID- 12571257 TI - Signaling molecules in nonfamilial pulmonary hypertension. AB - BACKGROUND: Biochemical, genetic, and clinical evidence indicates that smooth muscle proliferation around small pulmonary vessels is an essential part of the pathogenesis of pulmonary hypertension. Mutations in the bone morphogenetic protein receptor type 2 (BMPR2) have been linked to familial cases of pulmonary hypertension, but the molecular basis of the common nonfamilial forms is unknown. METHODS: We evaluated the pattern of expression of angiopoietin-1, a protein involved in the recruitment of smooth-muscle cells around blood vessels; TIE2, the endothelial-specific receptor for angiopoietin-1; and bone morphogenetic protein receptor type 1A (BMPR1A) and BMPR2 in lung-biopsy specimens from patients with pulmonary hypertension and from normotensive control patients. The effect of angiopoietin-1 on the modulation of BMPR expression was also evaluated in subcultures of human pulmonary arteriolar endothelial cells. RESULTS: The expression of angiopoietin-1 messenger RNA and the protein itself and the phosphorylation of TIE2 were strongly up-regulated in the lungs of patients with various forms of pulmonary hypertension, correlating directly with the severity of disease. A mechanistic link between familial and acquired pulmonary hypertension was demonstrated by the finding that angiopoietin-1 shuts off the expression of BMPR1A, a transmembrane protein required for BMPR2 signaling, in pulmonary arteriolar endothelial cells. Similarly, we found that the expression of BMPR1A was severely reduced in the lungs of patients with various forms of acquired as well as primary nonfamilial pulmonary hypertension. CONCLUSIONS: These findings suggest that all forms of pulmonary hypertension are linked by defects in the signaling pathway involving angiopoietin-1, TIE2, BMPR1A, and BMPR2 and consequently identify specific molecular targets for therapeutic intervention. PMID- 12571258 TI - Validation of a brief observation period for patients with cocaine-associated chest pain. AB - BACKGROUND: Retrospective studies of patients with cocaine-associated chest pain suggest that a strategy of discharging patients from the emergency department after a 12-hour observation period if they do not have evidence of ischemia should be associated with a very low rate of complications. METHODS: We prospectively evaluated the safety of a 9-to-12-hour observation period in patients with cocaine-associated chest pain who were at low-to-intermediate risk of cardiovascular events. Consecutive patients who reported or tested positive for cocaine use and who received protocol-driven care in a chest-pain observation unit were included. Patients who had normal levels of troponin I, without new ischemic changes on electrocardiography, and who had no cardiovascular complications (dysrhythmias, acute myocardial infarction, or recurrent symptoms) during the 9-to-12-hour observation period were discharged from the unit. The main outcome was death from cardiovascular causes at 30 days. RESULTS: Three hundred forty-four patients with cocaine-associated chest pain were evaluated. Forty-two of these patients (12 percent) were directly admitted to the hospital. The study cohort comprised the remaining 302 patients. During the 30-day follow up period, none of the patients died of a cardiovascular event (0 percent; 95 percent confidence interval, 0 to 0.99), and only 4 of the 256 patients for whom detailed follow-up data were available had a nonfatal myocardial infarction (1.6 percent; 95 percent confidence interval, 0.1 to 3.1). All four nonfatal myocardial infarctions occurred in patients who continued to use cocaine. CONCLUSIONS: Patients with cocaine-associated chest pain who do not have evidence of ischemia or cardiovascular complications over a 9-to-12-hour period in a chest pain observation unit have a very low risk of death or myocardial infarction during the 30 days after discharge. PMID- 12571259 TI - Epidemiologic classification of human papillomavirus types associated with cervical cancer. AB - BACKGROUND: Infection with human papilloma virus (HPV) is the main cause of cervical cancer, but the risk associated with the various HPV types has not been adequately assessed. METHODS: We pooled data from 11 case-control studies from nine countries involving 1918 women with histologically confirmed squamous-cell cervical cancer and 1928 control women. A common protocol and questionnaire were used. Information on risk factors was obtained by personal interviews, and cervical cells were collected for detection of HPV DNA and typing in a central laboratory by polymerase-chain-reaction-based assays (with MY09/MY11 and GP5+/6+ primers). RESULTS: HPV DNA was detected in 1739 of the 1918 patients with cervical cancer (90.7 percent) and in 259 of the 1928 control women (13.4 percent). With the GP5+/6+ primer, HPV DNA was detected in 96.6 percent of the patients and 15.6 percent of the controls. The most common HPV types in patients, in descending order of frequency, were types 16, 18, 45, 31, 33, 52, 58, and 35. Among control women, types 16, 18, 45, 31, 6, 58, 35, and 33 were the most common. For studies using the GP5+/6+ primer, the pooled odds ratio for cervical cancer associated with the presence of any HPV was 158.2 (95 percent confidence interval, 113.4 to 220.6). The odds ratios were over 45 for the most common and least common HPV types. Fifteen HPV types were classified as high-risk types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82); 3 were classified as probable high-risk types (26, 53, and 66); and 12 were classified as low-risk types (6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 81, and CP6108). There was good agreement between our epidemiologic classification and the classification based on phylogenetic grouping. CONCLUSIONS: In addition to HPV types 16 and 18, types 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82 should be considered carcinogenic, or high-risk, types, and types 26, 53, and 66 should be considered probably carcinogenic. PMID- 12571260 TI - Images in clinical medicine. Thoracic echinococcosis. PMID- 12571261 TI - Inheritance and drug response. PMID- 12571262 TI - Pharmacogenomics--drug disposition, drug targets, and side effects. PMID- 12571263 TI - Contrast-agent-induced acute renal dysfunction--is iodixanol the answer? PMID- 12571264 TI - Pharmacogenetics in the laboratory and the clinic. PMID- 12571265 TI - Extraction of an embolus. PMID- 12571266 TI - Hyperbaric oxygen for acute carbon monoxide poisoning. PMID- 12571267 TI - Aspirin, clopidogrel, or both for secondary prevention of coronary disease. PMID- 12571268 TI - RNA interference. PMID- 12571269 TI - Spinal cord disease in West Nile virus infection. PMID- 12571270 TI - Bicycle racing. PMID- 12571271 TI - Fatal melanoma transferred in a donated kidney 16 years after melanoma surgery. PMID- 12571272 TI - Images in clinical medicine. Aneurysm of the left anterior descending coronary artery. PMID- 12571273 TI - The trypanosome flagellum. PMID- 12571274 TI - Changing directions: clathrin-mediated transport between the Golgi and endosomes. AB - Clathrin-coated vesicles mediate transport between the trans-Golgi network (TGN) and endosomes. In recent years there has been tremendous progress in identifying factors involved in anterograde and retrograde transport steps. The well characterised heterotetrameric clathrin adaptor complex AP-1 has long been thought to mediate anterograde transport from the TGN to endosomes. However, recent studies of AP-1-knockout mice implicate AP-1 in retrograde as well as anterograde transport. The recently identified Golgi-associated, gamma-ear containing, ARF-binding (GGA) proteins share functional similarities with tetrameric adaptor complexes and are essential for anterograde transport of mannose-6-phosphate receptors, the sorting receptors for soluble lysosomal enzymes. To date, it is not clear whether GGAs and AP-1 mediate transport in different directions, act in parallel pathways, or cooperate in the same transport steps. Recent data have shed light on the locations, functions and interactions of AP-1 and GGA proteins. These data provide support for the role of both in anterograde transport from the Golgi complex. PMID- 12571275 TI - Prolyl isomerase Pin1: a catalyst for oncogenesis and a potential therapeutic target in cancer. AB - Phosphorylation of proteins on serine or threonine residues preceding proline (Ser/Thr-Pro) is a major intracellular signaling mechanism. The phosphorylated Ser/Thr-Pro motifs in a certain subset of phosphoproteins are isomerized specifically by the peptidyl-prolyl cis-trans isomerase Pin1. This post phosphorylation isomerization can lead to conformational changes in the substrate proteins and modulate their functions. Pin1 interacts with a number of mitotic phosphoproteins, and plays a critical role in mitotic regulation. Recent work indicates that Pin1 is overexpressed in many human cancers and plays an important role in oncogenesis. Pin1 regulates the expression of cyclin D1 by cooperating with Ras signaling and inhibiting the interaction of beta-catenin with the tumor suppressor APC and also directly stabilizing cyclin D1 protein. Furthermore, PIN1 is an E2F target gene essential for the Neu/Ras-induced transformation of mammary epithelial cells. Pin1 is also a critical regulator of the tumor suppressor p53 during DNA damage response. Given its role in cell growth control and oncogenesis, Pin1 could represent a new anti-cancer target. PMID- 12571276 TI - Local force induced conical protrusions of phagocytic cells. AB - Magnetic tweezers were used to study the passive and active response of macrophages to local centripetal nanonewton forces on beta1 integrins. Superparamagnetic beads coated with the beta1-integrin-binding protein invasin were attached to J774 murine macrophages to mimic phagocytosis of bacterial pathogens. Forces exceeding approximately 0.5 nN induce the active formation of trumpet-like protrusions resembling pseudopodia after an initial elastic deflection and a response time of approximately 30 seconds. The speed of advancement of the protrusion is =0.065+/-0.020 micro m second(-1) and is force independent. After saturation (after about 100 seconds) the protrusion stops abruptly and is completely retracted again against forces exceeding 5 nN with an effective relaxation time of approximately 30 seconds. The active protrusion is tentatively attributed to the growth of the actin cortex in the direction of the force, and evidence for the involvement of actin is provided by the finding that Latrunculin A abolishes the activated cone growth. The growth is assumed to be activated by cell signaling mediated by the invasin-specific integrins (exhibiting beta1 chains) and could play a role in phagocytic and protrusive events during immune response by macrophages. PMID- 12571277 TI - The Arabidopsis lue1 mutant defines a katanin p60 ortholog involved in hormonal control of microtubule orientation during cell growth. AB - The lue1 mutant was previously isolated in a bio-imaging screen for Arabidopsis mutants exhibiting inappropriate regulation of an AtGA20ox1 promoter-luciferase reporter fusion. Here we show that lue1 is allelic to fra2, bot1 and erh3, and encodes a truncated katanin-like microtubule-severing protein (AtKSS). Complementation of lue1 with the wild-type AtKSS gene restored both wild-type stature and luciferase reporter levels. Hormonal responses of lue1 to ethylene and gibberellins revealed inappropriate cortical microtubule reorientation during cell growth. Moreover, a fusion between the AtKSS protein and GFP decorated cortical microtubules. A yeast two-hybrid screen with AtKSS as the bait identified proteins related to those involved in microtubule processing, including a katanin p80 subunit and a kinesin ortholog. These results indicate that AtKSS is involved in microtubule dynamics in response to plant hormones. PMID- 12571278 TI - Expression of the nidogen-binding site of the laminin gamma1 chain disturbs basement membrane formation and maintenance in F9 embryoid bodies. AB - Basement membranes contain two major molecular networks consisting of laminin and collagen IV. Previous antibody perturbation experiments suggest that the interaction between laminin and nidogen-1 is necessary for proper basement membrane formation and epithelial development, whereas results from gene ablation experiments in mice show that both basement membranes and general development are grossly normal in the absence of nidogen-1. To refine the perturbation approach, we produced F9-teratocarcinoma-cell-derived embryoid bodies in the presence of recombinantly expressed nidogen-binding sites localized within the gamma1III3-5 laminin fragment. We found basement membranes were disrupted in gamma1III3-5 expressing embryoid bodies. As a measurement of basement membrane function, we tested permeability and detected drastically increased diffusion rates in correlation with basement membrane disruption. Furthermore, TROMA-1 localization in embryoid bodies expressing the nidogen-binding site was altered, suggesting separation of epithelium-specific gene expression from the formation of the actual epithelium when occurring in the absence of an organized basement membrane. PMID- 12571279 TI - Microtubule-disruption-induced and chemotactic-peptide-induced migration of human neutrophils: implications for differential sets of signalling pathways. AB - Neutrophil granulocytes rely on a functional actin network for directed migration. Microtubule disassembly does not impair receptor-linked chemotaxis, instead it induces development of polarity and chemokinesis in neutrophils concomitant with polarized distribution of alpha-actinin and F-actin. Cells stimulated with colchicine, which disassembles microtubules, migrate with a speed comparable to cells exposed to chemotactic peptide. We investigated signalling pathways involved in colchicine-induced neutrophil polarization and migration. Colchicine-induced development of polarity was insensitive to treatment with pertussis toxin, in contrast to chemotactic-peptide-induced shape changes, which were completely abolished by this treatment. Thus, colchicine does not appear to act via activating heterotrimeric G(i) proteins. Colchicine does also not seem to act via phosphatidylinositol 3-kinase, as it failed to induce phosphorylation of its downstream target Akt and the potent phosphatidylinositol 3-kinase inhibitor wortmannin failed to inhibit colchicine-induced shape changes. By contrast, wortmannin significantly reduced chemotactic-peptide-induced shape changes. However, the Rho-kinase inhibitor Y-27632 (10 micro M) inhibited colchicine induced development of polarity by 95+/-3% (n=5) and chemokinesis by 76+/-9% (n=3), which suggests that the Rho-Rho-kinase pathway has a crucial role in polarity and migration. Indeed, treatment of cells with colchicine induced a significant increase in membrane-bound Rho-kinase II, which is indicative of activation of this protein. This membrane translocation could be prevented by taxol, which stabilizes microtubules. Colchicine also induced a marked increase in myosin light chain phosphorylation, which could be suppressed by Y-27632 and by taxol. In summary, we provide evidence that microtubule disassembly induces in neutrophils a selective activation of Rho-kinase, bypassing activation of heterotrimeric Gi proteins and phosphatidylinositol 3-kinase. This process is sufficient for induction of chemokinesis and mediates increased phosphorylation of myosin light chain and accumulation of F-actin and alpha-actinin in the leading edge. PMID- 12571280 TI - Nonerythroid alphaII spectrin is required for recruitment of FANCA and XPF to nuclear foci induced by DNA interstrand cross-links. AB - The events responsible for repair of DNA interstrand cross-links in mammalian cells, the proteins involved and their interactions with each other are poorly understood. The present study demonstrates that the structural protein nonerythroid alpha spectrin (alphaSpIISigma*), present in normal human cell nuclei, plays an important role in repair of DNA interstrand cross-links. These results show that alphaSpIISigma* relocalizes to nuclear foci after damage of normal human cells with the DNA interstrand cross-linking agent 8-methoxypsoralen plus ultraviolet A (UVA) light and that FANCA and the known DNA repair protein XPF localize to the same nuclear foci. That alphaSpIISigma* is essential for this re-localization is demonstrated by the finding that in cells from patients with Fanconi anemia complementation group A (FA-A), which have decreased ability to repair DNA interstrand cross-links and decreased levels of alphaSpIISigma*, there is a significant reduction in formation of damage-induced XPF as well as alphaSpIISigma* nuclear foci, even though levels of XPF are normal in these cells. In corrected FA-A cells, in which levels of alphaSpIISigma* are restored to normal, numbers of damage-induced nuclear foci are also returned to normal. Co immunoprecipitation studies show that alphaSpIISigma*, FANCA and XPF co immunoprecipitate with each other from normal human nuclear proteins. These results demonstrate that alphaSpIISigma*, FANCA and XPF interact with each other in the nucleus and indicate that there is a close functional relationship between these proteins. These studies suggest that an important role for alphaSpIISigma* in the nucleus is to act as a scaffold, aiding in recruitment and alignment of repair proteins at sites of damage. PMID- 12571281 TI - Membrane insertion of the chloroplast outer envelope protein, Toc34: constrains for insertion and topology. AB - The insertion of the outer envelope protein Toc34 from chloroplasts was studied. Toc34 was chosen as a model protein because it contains one predicted transmembrane helix at the C-terminus and a large hydrophilic N-terminal located GTPase domain, which is exposed to the cytosol. Unlike proteins located in internal chloroplast compartments, Toc34 neither contains a cleavable presequence nor uses the general import pathway. The protein can insert into the outer envelope of chloroplasts but not into the outer membrane of mitochondria. Using protein-free liposomes we showed that Toc34 is able to insert directly into the lipid bilayer. This insertion is stimulated by GTP and the presence of nonbilayer lipids, but is independent of the presence or absence of charged lipids. The topology of the protein inserted into protein-free liposomes was not exclusively directed by the positive-inside rule but by the size of the hydrophilic domain. PMID- 12571282 TI - Real-time analysis of clathrin-mediated endocytosis during cell migration. AB - Simultaneous dual-color total-internal-reflection fluorescence microscopy (TIR FM) was performed to analyze the internalization and distribution of markers for clathrin-mediated endocytosis (clathrin, dynamin1, dynamin2 and transferrin) in migrating cells. In MDCK cells, which endogenously express dynamin2, the dynamin2 EGFP fluorescence demonstrated identical spatial and temporal behavior as clathrin both prior to and during internalization. By contrast, in the same cells, the neuronal dynamin1 only localized with clathrin just prior to endocytosis. In migrating cells, each endocytic marker was polarized towards the leading edge, away from the lagging edge. These observations suggest a re evaluation of the functional differences between dynamin1 and dynamin2, and of the role of clathrin-mediated endocytosis in cell migration. PMID- 12571283 TI - Mechanical loading regulates the expression of tenascin-C in the myotendinous junction and tendon but does not induce de novo synthesis in the skeletal muscle. AB - Tenascin-C is a hexabrachion-shaped matricellular protein with a very restricted expression in normal musculoskeletal tissues, but it is expressed abundantly during regenerative processes of these tissues and embryogenesis. To examine the importance of mechanical stress for the regulation of tenascin-C expression in the muscle-tendon unit, the effects of various states of mechanical loading (inactivity by cast-immobilization and three-varying intensities of subsequent re activity by treadmill running) on the expression of tenascin-C were studied using immunohistochemistry and mRNA in situ hybridization at the different locations of the muscle-tendon unit of the rat gastrocnemius muscle, the Achilles tendon complex. This muscle-tendon unit was selected as the study site, because the contracting activity of the gastrocnemius-soleus muscle complex, and thus the mechanical loading-induced stimulation, is easy to block by cast immobilization. Tenascin-C was expressed abundantly in the normal myotendinous and myofascial junctions, as well as around the cells and the collagen fibers of the Achilles tendon. Tenascin-C expression was not found in the normal skeletal muscle, although it was found in blood vessels within the muscle tissue. Following the removal of the mechanical loading-induced stimulation on the muscle-tendon unit by cast immobilization for 3 weeks, the immonoreactivity of tenascin-C substantially decreased or was completely absent in the regions expressing tenascin-C normally. Restitution of the mechanical loading by removing the cast and allowing free cage activity for 8 weeks resulted in an increase in tenascin-C expression, but it could not restore the expression of tenascin-C to the normal level (in healthy contralateral leg). In response to the application of a more strenuous mechanical loading stimulus after the removal of the cast (after 8 weeks of low- and high-intensity treadmill running), the expression of tenascin-C was markedly increased and reached the level seen in the healthy contralateral limb. Tenascin-C was abundantly expressed in myotendinous and myofascial junctions and in the Achilles tendon, but even the most strenuous mechanical loading (high-intensity treadmill running) could not induce the expression of tenascin-C in the skeletal muscle. This was in spite of the marked immobilization induced atrophy of the previously immobilized skeletal muscle, which had been subjected to intensive stress during remobilization. mRNA in situ hybridization analysis confirmed the immunohistochemical results for the expression of tenascin C in the study groups. In summary, this study shows that mechanical loading regulates the expression of tenascin-C in an apparently dose-dependent fashion at sites of the muscle-tendon unit normally expressing tenascin-C but can not induce de novo synthesis of tenascin-C in the skeletal muscle without accompanying injury to the tissue. Our results suggest that tenascin-C provides elasticity in mesenchymal tissues subjected to heavy tensile loading. PMID- 12571284 TI - Sterol-rich plasma membrane domains in the fission yeast Schizosaccharomyces pombe. AB - Sterol-rich membrane domains exist in unicellular and multicellular eukaryotes. They are thought to provide a structural framework for interactions among a subset of proteins by selectively incorporating some proteins while excluding others. Although most studies have focused on the biophysical and biochemical properties of sterol-rich membrane domains and incorporated proteins, relatively little is known about their intracellular distribution. Using a cytological approach we show here that in the fission yeast Schizosaccharomyces pombe, sterols are enriched in the plasma membrane at the growing cell tips and at the site of cytokinesis. The distribution of sterols is regulated in a cell-cycle dependent manner and requires a functional secretory pathway. By manipulating the integrity of sterol-rich membrane domains using sterol sequestering agents and genetic means, we show that these domains are important for multiple processes regulating cytokinesis. In these cells, defects in proper maintenance of the actomyosin ring and/or its attachment to the overlying plasma membrane were observed. Furthermore, the stability of a plasma membrane protein that colocalises with sterol-rich membrane domains was compromised. Taken together, our studies establish S. pombe as a genetically tractable model organism in which to study the role(s) of sterol-rich membrane domains in cell polarity and cytokinesis. PMID- 12571285 TI - The organization of adherens junctions and desmosomes at the cardiac intercalated disc is independent of gap junctions. AB - Adherens junctions and desmosomes are responsible for mechanically coupling myocytes in the heart and are found closely apposed to gap junction plaques at the intercalated discs of cardiomyocytes. It is not known whether loss of cardiac gap junctions, such as described in cardiac disease states, may influence the expression patterns of other intercalated disc-associated proteins. We investigated whether the major cardiac gap junction protein connexin43 (Cx43) may be responsible for regulating adherens junctions, desmosomes and their associated catenins, in terms of abundance and localization at the intercalated discs of cardiomyocytes. In order to study the effect of loss of cardiac gap junctions on the intercalated disc-associated proteins, we used a combination of immunoblotting, immunofluorescence with confocal microscopy and electron microscopy to evaluate heart tissue from mice with cardiac-specific conditional knockout of Cx43. We found that the cardiac adherens junctions, desmosomes and their associated catenins, as well as vinculin and ZO-1, maintain their normal abundance, structural appearance and localization in the absence of Cx43. We conclude from these data that Cx43 is not required for the organization of the cell adhesion junctions and their associated catenins at the intercalated disc in the adult cardiac myocyte. PMID- 12571286 TI - A mouse homologue of Drosophila pins can asymmetrically localize and substitute for pins function in Drosophila neuroblasts. AB - Asymmetric cell division is a fundamental mechanism used to generate cellular diversity in invertebrates and vertebrates. In Drosophila, asymmetric division of neuroblasts is achieved by the asymmetric segregation of cell fate determinants Prospero and Numb into the basal daughter cell. Asymmetric segregation of cell fate determinants requires an apically localized protein complex that includes Inscuteable, Pins, Bazooka, DmPar-6, DaPKC and Galphai. Pins acts to stabilize the apical complex during neuroblast divisions. Pins interacts and colocalizes with Inscuteable, as well as maintaining its apical localization. We have isolated a mouse homologue of pins (Pins) and characterized its expression profile. Mouse PINS shares high similarity in sequence and structure with Pins and other Pins-like proteins from mammals. Pins is expressed in many mouse tissues but its expression is enriched in the ventricular zone of the developing central nervous systems. PINS localizes asymmetrically to the apical cortex of mitotic neuroblasts when ectopically expressed in Drosophila embryos. Like Pins, its N-terminal tetratricopeptide repeats can directly interact with the asymmetric localization domain of Insc, and its C-terminal GoLoco-containing region can direct localization to the neuroblast cortex. We further show that Pins can fulfill all aspects of pins function in Drosophila neuroblast asymmetric cell divisions. Our results suggest a conservation of function between the fly and mammalian Pins homologues. PMID- 12571287 TI - TES is a novel focal adhesion protein with a role in cell spreading. AB - Previously, we identified TES as a novel candidate tumour suppressor gene that mapped to human chromosome 7q31.1. In this report we demonstrate that the TES protein is localised at focal adhesions, actin stress fibres and areas of cell cell contact. TES has three C-terminal LIM domains that appear to be important for focal adhesion targeting. Additionally, the N-terminal region is important for targeting TES to actin stress fibres. Yeast two-hybrid and biochemical analyses yielded interactions with several focal adhesion and/or cytoskeletal proteins including mena, zyxin and talin. The fact that TES localises to regions of cell adhesion suggests that it functions in events related to cell motility and adhesion. In support of this, we demonstrate that fibroblasts stably overexpressing TES have an increased ability to spread on fibronectin. PMID- 12571288 TI - Remodeling of endosomes during lysosome biogenesis involves 'kiss and run' fusion events regulated by rab5. AB - The small GTPase rab5 has been shown to play key roles in the function of both endocytic and phagocytic organelles. Although these organelles share several additional common features, different processes have been proposed to explain their biogenesis. In the present study, we provide evidence that lysosome biogenesis involves mechanisms similar to those previously described for the formation of phagolysosomes. Transient interactions ('kiss and run') between endocytic organelles are shown to occur during lysosome biogenesis. These interactions are regulated initially by the GTPase activity of rab5, as demonstrated by the loss of size-selective fusion between endosomes in cells expressing a GTPase-deficient mutant of rab5. Endocytic compartments in these cells sequentially display properties of early and late endosomes. However, the formation of lysosomes and the sorting of endocytic solute materials to small electron dense vacuoles are not affected by the rab5 mutation. Together, our results indicate that endosome maturation occurs during the early part of lysosome biogenesis. This process involves transient fusion events regulated, in part, by the small GTPase rab5. PMID- 12571289 TI - Non-membranous granular organelle consisting of PCM-1: subcellular distribution and cell-cycle-dependent assembly/disassembly. AB - Centriolar satellites were initially identified as electrondense spherical granules, approximately 70-100 nm in diameter, localized around the centrosomes. We have previously identified pericentriolar material 1 (PCM-1), with a molecular mass of approximately 230 kDa, as a component of centriolar satellites. We now show by immunofluorescence microscopy that these granules are not only concentrated around centrioles but also scattered throughout the cytoplasm in various types of mouse cells, leading us tentatively to call them 'PCM-1 granules'. We then found that, when overexpressed, PCM-1 molecules lacking their C-terminal region bound directly with each other through two distinct regions to form large aggregates, which then recruited endogenous PCM-1. These large aggregates as well as endogenous PCM-1 granules appear to be disassembled during mitosis, and reassembled when the cells entered interphase. These findings suggest that PCM-1 granules are formed by self-aggregation of PCM-1 and that this self-aggregation is regulated in a cell-cycle-dependent manner. Furthermore, we found that PCM-1 granules are distinct from pericentrin-containing granules, and that these two distinct types of granular structures are frequently associated with each other within the cytoplasm. These findings are discussed with special reference to the possible physiological functions of PCM-1 granules. PMID- 12571290 TI - Drosophila dd4 mutants reveal that gammaTuRC is required to maintain juxtaposed half spindles in spermatocytes. AB - The weak spindle integrity checkpoint in Drosophila spermatocytes has revealed a novel function of the gamma-tubulin ring complex (gammaTuRC) in maintaining spindle bipolarity throughout meiosis. Bipolar and bi-astral spindles could form in Drosophila mutants for dd4, the gene encoding the 91 kDa subunit of gammaTuRC. However, these spindles collapsed around metaphase and began to elongate as if attempting anaphase B. The microtubules of the collapsing spindle folded back on themselves, their putative plus ends forming the focused apexes of biconical figures. Cells with such spindles were unable to undergo cytokinesis. A second type of spindle, monopolar hemi-spindles, also formed as a result of either spindle collapse at an earlier stage or failure of centrosome separation. Multiple centrosome-like bodies at the foci of hemi-spindles nucleated robust asters of microtubules in the absence of detectable gamma-tubulin. Time-lapse imaging revealed these to be intermediates that developed into cones, structures that also had putative plus ends of microtubules focused at their tips. Unlike biconical figures, however, cones seemed to contain a central spindle-like structure at their apexes and undergo cytokinesis. We conclude that spermatocytes do not need astral microtubules nucleated by opposite poles to intersect in order to form a central spindle and a cleavage furrow. PMID- 12571292 TI - Autologous chondrocyte implantation and osteochondral cylinder transplantation in cartilage repair of the knee joint. A prospective, comparative trial. AB - BACKGROUND: Current methods used to restore the joint surface in patients with localized articular cartilage defects include transplantation of an autologous osteochondral cylinder and implantation of autologous chondrocytes. The purpose of this study was to evaluate the clinical and histological outcomes of these two techniques. METHODS: We performed a prospective clinical study to investigate the two-year outcomes in forty patients with an articular cartilage lesion of the femoral condyle who had been randomly treated with either transplantation of an autologous osteochondral cylinder or implantation of autologous chondrocytes. Biopsy specimens from representative patients of both groups were evaluated with histological staining, immunohistochemistry, and scanning electron microscopy. RESULTS: According to the postoperative Lysholm score, the recovery after autologous chondrocyte implantation was slower than that after osteochondral transplantation at six months (p < or = 0.015), twelve months (p < or = 0.001), and twenty-four months (p < or = 0.012). On the basis of the Meyers score and the Tegner activity score, the results were equally good with the two methods two years after treatment. Histomorphological evaluation of biopsy specimens within two years after autologous chondrocyte implantation demonstrated a complete, mechanically stable resurfacing of the defect in all patients. The tissue consisted mainly of fibrocartilage, while localized areas of hyaline-like regenerative cartilage could be detected close to the subchondral bone. Although a gap remained at the site of the transplantation in all five biopsy specimens examined as long as two years after osteochondral cylinder transplantation, histomorphological analysis and scanning electron microscopy revealed no differences between the osteochondral transplants and the surrounding original cartilage. CONCLUSIONS: Both treatments resulted in a decrease in symptoms. However, the improvement provided by the autologous chondrocyte implantation lagged behind that provided by the osteochondral cylinder transplantation. Histologically, the defects treated with autologous chondrocyte implantation were primarily filled with fibrocartilage, whereas the osteochondral cylinder transplants retained their hyaline character, although there was a persistent interface between the transplant and the surrounding original cartilage. Limitations of our study included the small number of patients, the relatively short (two-year) follow-up, and the absence of a control group. PMID- 12571293 TI - Incidence of wrong-site surgery among hand surgeons. AB - BACKGROUND: Until recently, wrong-site surgery had received little attention and had been considered a random, infrequent event. In 1997, the American Academy of Orthopaedic Surgeons (AAOS) Task Force on Wrong-Site Surgery was formed to determine the incidence of wrong-site surgery and to initiate the "Sign Your Site" campaign. The purpose of our study was to determine the incidence of wrong site surgery among hand surgeons, elucidate surgeons' practice habits and measures taken to prevent its occurrence, and evaluate the effectiveness of the AAOS "Sign Your Site" campaign. METHODS: One thousand, five hundred and sixty active members of the American Society for Surgery of the Hand (ASSH) were polled by mail. Each member received a confidential twenty-nine-question survey. Nonrespondents were sent a second, identical survey. One thousand and fifty (67%) of the surgeons responded. RESULTS: One hundred and seventy-three surgeons (16%) reported that they had prepared to operate on the wrong site but then noticed the error prior to the incision, and 217 (21%) reported performing wrong-site surgery at least once. Of an estimated 6,700,000 surgical procedures, 242 were performed at the wrong site, an incidence of one in 27,686 procedures. The three most common locations of wrong-site surgery were the fingers (153), hands (twenty), and wrists (twenty-one). Permanent disability occurred in twenty-one patients (9%). Ninety-three cases (38%) led to legal action or monetary settlement. Seventy percent of the responding orthopaedic surgeons were aware of the "Sign Your Site" campaign, and 45% had changed their practice habits as a result. CONCLUSIONS: Prior to the AAOS "Sign Your Site" campaign, the issue of wrong-site surgery by hand surgeons had not been addressed. Although wrong-site surgery is rare, 21% of hand surgeons reported performing it at least once during their careers. Since the institution of the "Sign Your Site" campaign, 45% of orthopaedic hand surgeons have changed their practice habits, and almost all routinely take some action to prevent wrong-site surgery. PMID- 12571294 TI - Total hip arthroplasty in adult patients who had childhood infection of the hip. AB - BACKGROUND: Adult patients with a history of infection of the hip in childhood present a challenge as candidates for total hip arthroplasty because of abnormal bone development, soft-tissue contractures, the possibility of reinfection, and their relative youth. The purpose of this study was to analyze the mid-term results of primary total hip arthroplasty in adult patients who had had infection of the hip in childhood. METHODS: We retrospectively analyzed 170 total hip arthroplasties that had been performed between 1983 and 1996 in patients who had had infection of the hip during childhood. Forty-seven hips had cemented or hybrid total hip replacements, and 123 hips had cementless total hip replacements. The age of the patients at the time that the infection was contracted was an average of 7.3 years (range, one to eleven years). The average age of the patients at the time of the index total hip arthroplasty was 41.9 years in the cemented or hybrid group and 42.7 years in the cementless group. The interval between active infection and arthroplasty was 34.9 years in the cemented or hybrid group and 31.9 years in the cementless group. All but two hips (one patient) had a quiescent period of infection of more than ten years. The average duration of follow-up was 9.8 years (range, seven to seventeen years) in the cemented or hybrid group and 10.8 years (range, seven to seventeen years) in the cementless group. RESULTS: All hips with a quiescent period of more than ten years had no recurrence of infection. The remaining two hips in one patient with a quiescent period of seven years had recurrence of the infection. The mean Harris hip scores improved from 50 points preoperatively to 85 points at the latest follow-up examination in the cemented or hybrid group and from 50 points preoperatively to 89 points at the latest follow-up examination in the cementless group. Eight (17%) of forty-seven hips with cemented or hybrid fixation and eighteen (15%) of 123 hips with cementless fixation had revision of both components because of aseptic loosening and/or osteolysis. The mean rate of linear wear of the polyethylene was 0.25 mm per year in the cemented or hybrid group and 0.29 mm per year in the cementless group. The prevalence of osteolysis was 53% (twenty-five of forty-seven hips) in the cemented or hybrid group and 59% (seventy-two of 123 hips) in the cementless group. CONCLUSIONS: There was no recurrence of infection after total hip arthroplasty in the patients with a quiescent period of infection of more than ten years. These young, active patients with technically difficult arthroplasties were at considerable risk for aseptic loosening. The prevalence of polyethylene wear and osteolysis was attributable to the less than optimal prosthetic designs and materials used during the time-period of this study. PMID- 12571295 TI - Use of a cast compared with a functional ankle brace after operative treatment of an ankle fracture. A prospective, randomized study. AB - BACKGROUND: Controversy continues with regard to the optimal postoperative care after open reduction and internal fixation of an ankle fracture. The hypothesis of this study was that postoperative treatment of an ankle fracture with a brace that allows active and passive range-of-motion exercises would improve the functional recovery of patients compared with that after conventional treatment with a cast. Thus, the purpose of this prospective, randomized study was to compare the long-term subjective, objective, and functional outcome after conventional treatment with a cast and that after use of functional bracing in the first six weeks following internal fixation of an ankle fracture. METHODS: One hundred patients with an unstable and/or displaced Weber type-A or B ankle fracture were treated operatively and then were randomly allocated to two groups: immobilization in a below-the-knee cast (fifty patients) or early mobilization in a functional ankle brace (fifty patients) for the first six postoperative weeks. The follow-up examinations, which consisted of subjective and objective (clinical, radiographic, and functional) evaluations, were performed at two, six, twelve, and fifty-two weeks and at two years postoperatively. RESULTS: There were no perioperative complications in either study group, but eight patients who were managed with a cast and thirty-three patients who were managed with a brace had postoperative complications, which were mainly related to wound-healing. Two patients in the group treated with a cast had deep-vein thrombosis. All fractures healed well in both groups. The difference between the two groups with respect to the complication rate was significant (p = 0.0005). No significant differences between the study groups were observed in the final subjective or objective (clinical) evaluation. At the two-year follow-up examination, the average score (and standard deviation) according to the ankle-rating scale of Kaikkonen et al. was 85 +/- 9 points for the group treated with a cast and 83 +/- 10 points for the group treated with a brace, and the average ankle score according to the system of Olerud and Molander was 87 +/- 8 points and 87 +/- 9 points, respectively. CONCLUSIONS: The long-term functional outcome after postoperative treatment of an ankle fracture with a cast and that after use of a functional brace are similar. Although early mobilization with use of a functional ankle brace may have some theoretical beneficial effects, the risk of postoperative wound complications associated with this treatment approach is considerably increased compared with that after conventional cast treatment. Thus, the postoperative protocol of treatment with a functional brace requires refinement before it can be generally advocated for use after operative treatment of an ankle fracture. PMID- 12571296 TI - Patellar resurfacing in total knee arthroplasty. A prospective, randomized study. AB - BACKGROUND: Anterior knee pain following total knee arthroplasty is a common complaint and typically is attributed to the patellofemoral joint. The purpose of the present study was to compare the outcome of resurfacing and nonresurfacing of the patella, particularly with regard to anterior knee pain, and to clarify the indications for patellar resurfacing at the time of total knee arthroplasty. METHODS: We performed a prospective, randomized study of 514 consecutive primary press-fit condylar total knee replacements. The patients were randomized to either resurfacing or retention of the patella. They were also randomized to either a cruciate-substituting or a cruciate-retaining prosthesis as part of a separate trial. The mean duration of follow-up was 5.3 years (range, two to 8.5 years), and the patients were assessed with use of the Knee Society rating, a clinical anterior knee pain score, and the British Orthopaedic Association patient-satisfaction score. The assessment was performed without the examiner knowing whether the patella had been resurfaced. At the time of follow-up, there were 474 knees. Thirty-five patients who had a bilateral knee replacement underwent resurfacing on one side only. RESULTS: The overall prevalence of anterior knee pain was 25.1% (fifty-eight of 231 knees) in the nonresurfacing group, compared with 5.3% (thirteen of 243 knees) in the resurfacing group (p < 0.0001). There was one case of component loosening. Ten of eleven patients who underwent secondary resurfacing had complete relief of anterior knee pain. The overall postoperative knee scores were lower in the nonresurfacing group, and the difference was significant among patients with osteoarthritis (p < 0.01). There was no significant difference between the resurfacing and nonresurfacing groups with regard to the postoperative function score. Patients who had a bilateral knee replacement were more likely to prefer the resurfaced side. CONCLUSIONS: As the present study showed a significantly higher rate of anterior knee pain following arthroplasty without patellar resurfacing, we recommend patellar resurfacing at the time of total knee replacement when technically possible. PMID- 12571297 TI - Total knee arthroplasty in patients with a prior fracture of the tibial plateau. AB - BACKGROUND: A fracture of the tibial plateau may predispose the knee to the development of posttraumatic arthritis. Malunion, intra-articular chondro-osseous defects, limb malalignment, retained internal fixation devices, and poor surrounding soft tissues may in turn compromise the outcome of total knee arthroplasty. The aim of our study was to evaluate the results of total knee arthroplasty in patients with a previous fracture of the tibial plateau. METHODS: The results of sixty-two condylar total knee arthroplasties performed with cement, from 1988 to 1999, in sixty-two patients with a previous fracture of the tibial plateau were reviewed. The fracture of the tibial plateau had been treated by open reduction and internal fixation in thirty-eight knees, external fixation in one knee, and nonoperatively in twenty-three knees. There were forty women and twenty-two men with an average age of sixty-three years at the time of the arthroplasty. Knee Society scores were recorded preoperatively and at the time of follow-up, at an average of 4.7 years, and complications were noted. No patient was lost to follow-up. RESULTS: The mean Knee Society scores improved significantly (p < 0.0001), from 43.9 points for pain and 52 points for function preoperatively to 82.9 and 84 points, respectively, at the time of the latest follow-up. There were thirteen reoperations, which included manipulation with the patient under anesthesia (five knees), wound revision (three knees), and component revision (five knees). There were six intraoperative complications (10%). A postoperative complication occurred in sixteen knees (26%). CONCLUSIONS: The vast majority of patients treated with total knee arthroplasty after a previous fracture of the tibial plateau have substantial improvement in function and relief of pain. However, these patients are at increased risk for perioperative complications, as evidenced by the high reoperation rate of 21% in this study. PMID- 12571298 TI - Experimental tibial plateau fractures augmented with calcium phosphate cement or autologous bone graft. AB - BACKGROUND: Depression fractures of the tibial plateau are often managed with use of internal fixation and autologous bone-grafting to maintain an anatomical reduction. Bone-grafting, however, provides only limited stability. As calcium phosphate cements have appropriate mechanical properties, they may provide a more suitable alternative. The objective of this study was to compare the effect of a calcium phosphate cement with that of impacted cancellous autograft for maintaining an anatomical reduction in an experimental model of a tibial plateau fracture. METHODS: Standardized cylindrical subchondral defects that were 8 mm in diameter and 10 mm deep were created bilaterally beneath the subchondral bone of the articular cartilage in the lateral tibial plateau of goats. An osteotome was used to fracture the overlying subchondral plate and articular cartilage. The plateau fracture fragment was completely depressed into the subchondral defect and then was anatomically reduced. The defects were randomly filled with either calcium phosphate cement or cancellous autograft. No internal fixation was used. The tibiae were harvested at varying time-periods that ranged from twenty-four hours to eighteen months. The stiffness of the healing augmented plateau fractures was determined. Histological specimens were assigned a score for degenerative changes. Loss of anatomic reduction was demonstrated in photomicrographs, and the amount of subsidence of the osteochondral fragment was measured in whole-mount histological sections. RESULTS: The prevalence and degree of fracture subsidence was significantly reduced at all time-points in the defects treated with calcium phosphate cement compared with those filled with autograft (p < 0.05). There were no significant differences in fracture stiffness between the two treatment groups at any of the time-points examined. The calcium phosphate cement was rapidly resorbed, and the volume fraction of the calcium phosphate cement was decreased to 4% at six months. The trabecular bone volume in the defects was restored to that of the intact controls at six months in both treatment groups. CONCLUSIONS: Cancellous autograft did not maintain an anatomical reduction of the tibial plateau fractures in this model. In contrast, augmentation with calcium phosphate cement prevented subsidence of the fracture fragment and maintained articular congruency as the fracture healed. The improved articular congruency reduced the prevalence and severity of degenerative changes in the joint. PMID- 12571299 TI - Articular fractures of the distal part of the humerus. AB - BACKGROUND: The purpose of this retrospective study was to identify the patterns of distal humeral articular fractures and to analyze the results of open reduction and internal fixation of these injuries. METHODS: The cases of twenty one patients with an articular fracture of the distal part of the humerus were reviewed at an average of forty months after the injury. Five components of the injury were identified: (1) the capitellum and the lateral aspect of the trochlea, (2) the lateral epicondyle, (3) the posterior aspect of the lateral column, (4) the posterior aspect of the trochlea, and (5) the medial epicondyle. All fractures were reduced and were stabilized with implants buried beneath the articular surface. RESULTS: All fractures healed, and no patient had residual ulnohumeral instability or weakness. Ten patients required a second operation: six, for release of an elbow contracture; two, for treatment of ulnar neuropathy; one, for removal of hardware causing symptoms; and one, because of early loss of fixation. The average arc of ulnohumeral motion was 96 degrees (range, 55 degrees to 140 degrees ). The results according to the Mayo Elbow Performance Index were excellent in four patients, good in twelve, and fair in five. CONCLUSIONS: Apparent fractures of the capitellum are often more complex fractures of the articular surface of the distal part of the humerus. Treatment of these injuries with operative reduction and fixation with buried implants can result in satisfactory restoration of elbow function. PMID- 12571300 TI - Reduction of a dislocation of the hip due to developmental dysplasia. Implications for the need for future surgery. AB - BACKGROUND: Recent reports on the treatment of a dislocation of the hip due to developmental dysplasia have documented conflicting data on the importance of the ossific nucleus in the development of postreduction ischemic necrosis. Delaying reduction until the ossific nucleus is present bypasses the time-period of maximal osseous remodeling of the hip, thereby possibly increasing the need for future operations. We hypothesized that hips with an ossific nucleus are more likely to have subsequent reconstructive procedures. METHODS: A retrospective review of the medical records at two tertiary-care children's hospitals was completed to identify all patients who had had reduction of a dislocation of the hip due to developmental dysplasia, performed between 1979 and 1993, when they were less than two years old. Patients were excluded if the medical records or radiographs were inadequate, the duration of follow-up was less than three years after the final reduction, a previous reduction had been performed at an outside facility, or the patient had a neuromuscular disease or a teratologic dislocation. We identified 124 patients (153 hips) who satisfied the criteria for inclusion. The average age at the time of the reduction was eleven months, and the average duration of follow-up was 7.2 years. RESULTS: Overall, fourteen of the sixty-three hips without an ossific nucleus had a reconstructive procedure: thirteen had a varus rotational osteotomy of the proximal part of the femur and one had a combined pelvic and varus rotational femoral osteotomy. Forty of the ninety hips with an ossific nucleus had a reconstructive procedure: twenty-seven had a varus rotational osteotomy, eight had a pelvic osteotomy, and five had a combined pelvic and varus rotational osteotomy (p < 0.05). In addition, secondary reconstructive procedures were performed in 17% (ten) of the fifty-nine patients who were less than six months old and in 35% (thirty-three) of the ninety-four patients who were at least six months old, which was a greater than twofold increase. The effect of age was further emphasized at the other age cutoff points. CONCLUSION: Delaying the reduction of a dislocated hip until the appearance of the ossific nucleus more than doubles the need for future surgery to make the hip as anatomically normal as possible. Despite finding a slight increase in the rate of ischemic necrosis after reduction of the hips without an ossific nucleus, we advocate early reduction of a dislocation of a hip due to developmental dysplasia to optimize the development of the hip with the minimum number of operations PMID- 12571301 TI - A modified periacetabular osteotomy with use of the transtrochanteric exposure. AB - BACKGROUND: Periacetabular osteotomies are technically demanding surgical procedures. We developed a modified technique that uses a transtrochanteric approach. Our goal was to facilitate surgical exposure without compromising the results of surgery. The purpose of the present study was to review the early results in our initial group of patients who had this procedure. METHODS: We retrospectively reviewed the results of a modified periacetabular osteotomy in thirty-eight consecutive patients (forty-six hips) at an average duration of follow-up of 4.2 years. The average age of the patients was thirty-one years (range, eighteen to fifty-eight years) at the time of surgery. We evaluated the results with use of serial radiographs and the Merle d'Aubigne and Postel hip scoring system. RESULTS: All osteotomies healed. Preoperatively, the average lateral center-edge angle was 3 degrees (range, -15 degrees to 12 degrees ), the anterior center-edge angle was 0 degrees (range, -20 degrees to 5 degrees ), the acetabular angle of Sharp was 57 degrees (range, 50 degrees to 68 degrees ), and the femoral head coverage was 53% (range, 40% to 66%). Postoperatively, the average lateral center-edge angle was 35 degrees (range, 23 degrees to 45 degrees ), the anterior center-edge angle was 32 degrees (range, 20 degrees to 55 degrees ), the acetabular angle of Sharp was 40 degrees (range, 37 degrees to 45 degrees ), and the femoral head coverage was 92% (range, 85% to 100%). The average amount of medialization of the femoral head was 6 mm. At the latest follow-up examination, improvement in the grade of osteoarthritis was noted in eight hips, while progression of osteoarthritis was seen in five hips. Functionally, the average Merle d'Aubigne and Postel hip score improved from 13.2 points preoperatively to 17.0 points postoperatively. No patient had a neurovascular complication. CONCLUSION: Our early experience with a modified periacetabular osteotomy showed encouraging results in terms of the technical ease of the technique and the outcome in our patients. PMID- 12571302 TI - Influence of preoperative factors on outcome of shoulder arthroplasty for glenohumeral osteoarthritis. AB - BACKGROUND: The results of shoulder arthroplasty for osteoarthritis have been reported to be excellent or good for the majority of patients, but the value of using a glenoid component and the anatomic factors that affect outcome are still debated. The purpose of this study was to evaluate the influence of an operatively confirmed full-thickness tear of the rotator cuff, the severity of preoperative erosion of glenoid bone, preoperative radiographic evidence of subluxation of the humeral head, and the severity of preoperative loss of the passive range of motion on the outcome of total shoulder arthroplasty and hemiarthroplasty. METHODS: In a multicenter clinical outcome study, we evaluated 128 shoulders in 118 patients with primary osteoarthritis who had been followed for a mean of forty-six months (range, twenty-four to eighty-seven months). RESULTS: Patients with <10 degrees of passive external rotation preoperatively had significantly less improvement in external rotation after hemiarthroplasty (p = 0.006). Thirteen (10%) of the 128 shoulders had a repairable full-thickness tear of the supraspinatus tendon, but these tears did not affect the overall American Shoulder and Elbow Surgeons score, the decrease in pain, or patient satisfaction. Severe or moderate eccentric glenoid erosion was seen in twenty nine (23%) of the 128 shoulders, and total shoulder arthroplasty resulted in significantly better passive total elevation and active external rotation as well as a trend toward significantly better active forward flexion than did hemiarthroplasty in these shoulders. The humeral head was subluxated posteriorly in twenty-three shoulders (18%), and when they were compared with the other shoulders in the study, these shoulders were found to have lower final American Shoulder and Elbow Surgeons scores, more pain, and decreased active external rotation following either total shoulder arthroplasty or hemiarthroplasty. CONCLUSIONS: On the basis of our data, we recommend the use of a glenoid component in shoulders with glenoid erosion. Humeral head subluxation was associated with a less favorable result regardless of the type of shoulder arthroplasty and must be considered in preoperative planning and counseling. Severe loss of the passive range of motion preoperatively was associated with a decreased passive range of motion postoperatively. A repairable tear of the supraspinatus tendon is not a contraindication to the use of a glenoid component. . PMID- 12571303 TI - Factors affecting the durability of primary total knee prostheses. AB - BACKGROUND: Failure of total knee arthroplasty is problematic. The purpose of this study was to evaluate the factors that influence the durability of a primary total knee prosthesis. METHODS: A survivorship analysis of 11,606 primary total knee arthroplasties carried out between January 1, 1978, and December 31, 2000, was performed. An analysis of patient and implant-related factors affecting survivorship was done with use of a multivariate Cox model. RESULTS: The survivorship was 91% (95% confidence interval, 90% to 91%) at ten years (2943 knees), 84% (95% confidence interval, 82% to 86%) at fifteen years (595 knees), and 78% (95% confidence interval, 74% to 81%) at twenty years (104 knees) following the surgery. Prosthetic survivorship at ten years was 83% for patients fifty-five years of age or less compared with 94% for those older than seventy years of age (p < 0.0001), 90% for those with a diagnosis of osteoarthritis compared with 95% for those with inflammatory arthritis (p < 0.005), and 91% for those with retention of the posterior cruciate ligament compared with 76% for those with substitution of the posterior cruciate ligament (a posterior stabilized prosthesis) (p < 0.0001). Survivorship at ten years was 92% for nonmodular metal-backed tibial components, 90% for modular metal-backed tibial components, and 97% for all-polyethylene tibial components (p < 0.0001). Survivorship at ten years was 92% for prostheses fixed with cement compared with 61% for those fixed without cement (p < 0.0001). CONCLUSIONS: Significant risk factors for failure of total knee arthroplasty were the type of implant, age and gender of the patient, diagnosis, type of fixation, and design of the patellar component. In the ideal situation-treatment of a woman over the age of seventy years who has inflammatory arthritis with a nonmodular, metal-backed tibial component, cement fixation, an all-polyethylene patellar component, and retention of the posterior cruciate ligament-the ten-year survivorship of the prosthesis was estimated to be 98% (95% confidence interval, 97% to 99%). PMID- 12571304 TI - Rotational acetabular osteotomy in patients forty-six years of age or older: comparison with younger patients. AB - BACKGROUND: Satisfactory intermediate and long-term results of periacetabular rotational osteotomy for early osteoarthritis secondary to dysplasia of the hip have been reported for patients in the third and fourth decades of life. The purpose of the present study was to examine the usefulness of rotational acetabular osteotomy in patients older than forty-six years of age. METHODS: A retrospective review of two groups of patients who had been treated with a rotational acetabular osteotomy was conducted. The older group consisted of twenty-four patients (twenty-six hips) with early-stage osteoarthritis who had a mean age at the time of surgery of 50.9 years (range, forty-six to fifty-eight years) and a mean duration of follow-up of 8.2 years (range, five to thirteen years), and the younger group consisted of sixty patients (sixty-three hips) who had a mean age at the time of surgery of 34.4 years (range, thirteen to forty five years) and a mean duration of follow-up of 8.3 years (range, five to fourteen years). Clinical follow-up was based on the system of Merle d'Aubigne and Postel. The center-edge angle, acetabular roof angle, and head lateralization index were measured on radiographs made preoperatively, postoperatively, and at the time of follow-up. Preoperative and postoperative joint congruencies were classified into four grades. RESULTS: Preoperatively, the mean Merle d'Aubigne clinical score was 13.9 points in the older group and 14.1 points in the younger group. In both groups, this score improved significantly to a mean postoperative follow-up score of 16.6 points (p < 0.0001), with no significant difference between the two groups. The mean center-edge angle improved from 3.2 degrees preoperatively to 34 degrees postoperatively (p < 0.0001) in the older group and from -2.1 degrees preoperatively to 34 degrees postoperatively (p < 0.0001) in the younger group. The mean acetabular roof angle improved from 29 degrees to 5.9 degrees (p < 0.0001) in the older group and from 31 degrees to 2.9 degrees (p < 0.0001) in the younger group. The mean head lateralization index improved from 0.67 to 0.64 (p < 0.01) in the older group and from 0.66 to 0.61 (p < 0.0001) in the younger group. Progression of osteoarthritis was observed radiographically at the time of follow-up in five hips in the older group and in four hips in the younger group. Kaplan-Meier survivorship analysis, with radiographic progression of osteoarthritis as the end point, predicted a ten-year survival rate of 70.0% in the older group and 93.7% in the younger group; this difference was not significant, with the numbers available (p = 0.062, log-rank test). CONCLUSIONS: Our results indicate that rotational acetabular osteotomy for elderly patients can prevent progression of osteoarthritis (as indicated by a survival rate of 70% at ten years) and that in selected cases it is worthwhile at least as a temporizing operation. PMID- 12571305 TI - Combined vertical and horizontal cable fixation of an extended trochanteric osteotomy site. AB - BACKGROUND: The use of an extended trochanteric osteotomy facilitates exposure and aids in the removal of a well-fixed femoral implant and cement during revision total hip arthroplasty. Occasionally, nonunion, fracture, and trochanteric migration have been reported following osteotomy. We evaluated the rate of healing of the osteotomy site and of implant stability when fixation was accomplished with use of vertical trochanteric and horizontal metaphyseal cable fixation (combined cable fixation). METHODS: The clinical and radiographic results of revision total hip arthroplasty with use of an extended trochanteric osteotomy followed by implantation of a distally porous-coated component and combined cable fixation of the osteotomy site in forty-two consecutive patients (forty-three hips) were reviewed. Intraoperative fracture at the osteotomy site occurred in five hips (12%). RESULTS: All osteotomy sites healed by six months, with an average time to union of fifteen weeks. One implant subsided 5 mm in a patient in whom a fracture had occurred at the time of the osteotomy. No trochanteric migration occurred. Two patients required a reoperation: one because of instability, and another because of recurrent infection. CONCLUSIONS: The extended trochanteric osteotomy facilitates revision of a well-fixed femoral component. Despite occasional intraoperative fracture at the osteotomy site, combined vertical trochanteric and horizontal metaphyseal cable fixation resulted in an excellent rate of healing and implant stability. PMID- 12571306 TI - Anterior femoro-acetabular impingement due to acetabular retroversion. Treatment with periacetabular osteotomy. AB - BACKGROUND: This study was performed to evaluate whether symptomatic anterior femoro-acetabular impingement due to acetabular retroversion can be treated effectively with a periacetabular osteotomy. METHODS: The diagnosis of femoro acetabular impingement was based on clinical symptoms, a positive anterior impingement test, and findings of acetabular rim lesions on magnetic resonance imaging. The radiographic diagnosis of acetabular retroversion was based on the cross-over and posterior wall signs. Twenty-nine hips in twenty-two patients (average age, twenty-three years) underwent a periacetabular osteotomy. An arthrotomy was performed in twenty-six hips in order to visualize intra-articular lesions and, in selected cases, to improve a low femoral head-neck offset. The range of motion of the hip was measured, clinical evaluation was performed with use of the score described by Merle d'Aubigne and Postel, and the anterior center edge angle of Lequesne and de Seze was measured on radiographs preoperatively and at the time of the latest follow-up. RESULTS: The duration of follow-up averaged thirty months (range, twenty-four to forty-nine months). The anterior center-edge angle of Lequesne and de Seze decreased significantly from a preoperative average of 36 degrees (range, 26 degrees to 52 degrees ) to a postoperative average of 28 degrees (range, 16 degrees to 46 degrees ) (p = 0.002). There was a significant increase in the average range of internal rotation (10 degrees, p = 0.006), flexion (7 degrees, p = 0.014), and adduction (8 degrees, p = 0.017). The average Merle d'Aubigne score increased from 14.0 points (range, 12 to 16 points) preoperatively to 16.9 points (range, 15 to 18 points) postoperatively (p < 0.001), and the result was good or excellent for twenty-six hips. Three hips underwent subsequent surgery: one, because of early postoperative loss of reduction; one, for correction of posteroinferior impingement; and one, because of recurrent signs of anterior impingement. CONCLUSION: Periacetabular osteotomy is an effective way to reorient the acetabulum in young adults with symptomatic anterior femoro-acetabular impingement due to acetabular retroversion PMID- 12571307 TI - Tibial plafond fractures. How do these ankles function over time? AB - BACKGROUND: The intermediate outcome of fractures of the tibial plafond treated with current techniques has not been reported, to our knowledge. The purpose of this study, performed at a minimum of five years after injury, was to determine the effect of these fractures on ankle function, pain, and general health status and to determine which factors predict favorable and unfavorable outcomes. METHODS: Fifty-six ankles (fifty-two patients) with a tibial plafond fracture were treated with a uniform technique consisting of application of a monolateral hinged transarticular external fixator coupled with screw fixation of the articular surface. Thirty-one patients with thirty-five involved ankles returned between five and twelve years after the injury for a physical examination, assessment of ankle pain and function with the Iowa Ankle Score and Ankle Osteoarthritis Scale, assessment of general health status with the Short Form-36 (SF-36), and radiographic examination of the ankle. RESULTS: Arthrodesis had been performed on five of the forty ankles for which the outcome was known at a minimum of five years after the injury. Other than removal of prominent screws (two patients), no other surgical procedure had been performed on any patient. The average Iowa Ankle Score was 78 points (range, 28 to 96 points). The scores on the SF-36 and Ankle Osteoarthritis Scale demonstrated a long-term negative effect of the injury on general health and on ankle pain and function when compared with those parameters in age-matched controls. The degree of osteoarthrosis was grade 0 in three ankles, grade 1 in six, grade 2 in twenty, and grade 3 in six. The majority of patients had some limitation with regard to recreational activities, with an inability to run being the most common complaint (twenty-seven of the thirty-one patients). Fourteen patients changed jobs because of the ankle injury. Fifteen ankles were rated by the patient as excellent; ten, as good; seven, as fair; and one, as poor. Nine patients with previously recorded ankle scores had better scores after the longer follow-up interval. The patients perceived that their condition had improved for an average of 2.4 years after the injury. CONCLUSIONS: Although tibial plafond fractures have an intermediate-term negative effect on ankle function and pain and on general health, few patients require secondary reconstructive procedures and symptoms tend to decrease for a long time after healing. PMID- 12571308 TI - Total hip arthroplasty with use of the cementless Zweymuller Alloclassic system. A ten to thirteen-year follow-up study. AB - BACKGROUND: The Zweymuller Alloclassic total hip arthroplasty system is widely used, although few intermediate-term studies have been published. The purpose of the present study was to evaluate the clinical and radiographic results of this system after ten years. METHODS: One hundred and twenty-four consecutive primary total hip arthroplasties were performed with the Zweymuller Alloclassic cementless system at our institution between February 1988 and March 1991, and 104 hips (ninety-four patients) were retrospectively reviewed after a minimum duration of follow-up of ten years. The mean age of the patients at the time of the arthroplasty was 62.3 years (range, twenty-five to seventy-seven years). The mean duration of follow-up was 11.3 years. Standard radiographs were made for all patients immediately after the operation, at six and twelve months, and annually thereafter for at least ten years. Cox multivariate regression analysis was performed to assess the influence of various factors on survival of the implant. RESULTS: The cumulative probability of not having a revision of any prosthetic component for any reason was 94.1% (95% confidence interval, 91.9% to 96.3%) at twelve years in the best-case scenario and 85.3% (95% confidence interval, 82.1% to 88.5%) at twelve years in the worst-case scenario for the entire series of 124 hips. Among the 104 hips in the follow-up study, three acetabular components and no stems were revised. Two hips had level-4 pain according to the system of Merle D'Aubigne and Postel. Seven acetabular components (7%) were loose at twelve years, and all stems had radiographic evidence of stable fixation. Acetabular cup loosening was related to a vertical cup angle (p = 0.0008, Student t test), acetabular wear of > or =1 mm (p = 0.001, Fisher exact test), and a 32-mm femoral head (p = 0.001, Fisher exact test). Although femoral osteolysis was seen in eighteen hips (17%) at twelve years, all osteolytic cavities were proximal and focal. CONCLUSIONS: The Zweymuller Alloclassic prosthesis, particularly its femoral stem, demonstrated good results and durable fixation at a minimum of ten years of follow-up. PMID- 12571309 TI - Humeral fixation by press-fitting of a tapered metaphyseal stem: a prospective radiographic study. AB - BACKGROUND: The technique of shoulder arthroplasty requires a method for securely and durably fixing the humeral component in the medullary canal of the proximal part of the humerus. As an alternative to fixation by cementing or tissue ingrowth, we explored the effectiveness of using a prosthesis with a metaphyseal taper from the anatomic neck to the diaphysis. This component is designed to obtain press-fit fixation in the cancellous bone of the metaphysis rather than in the cortical bone of the diaphysis. We tested the hypothesis that this press-fit humeral stem, designed to respect the taper of the proximal part of the humeral canal, would be associated with a low rate of loosening in patients managed with primary shoulder arthroplasty for osteoarthritis. METHODS: A prospective study was conducted to evaluate the prevalence of radiolucent lines around press-fit humeral prostheses. One hundred and thirty-one patients with glenohumeral osteoarthritis were followed for a minimum of two years. A zonal method of evaluating radiolucent lines was established. Shift in position and subsidence were judged qualitatively. RESULTS: No component showed subsidence or a shift in position. Fifty shoulders (39%) had no radiolucency. Two shoulders had radiolucency around the proximal part of the prosthesis, and seventy-five had radiolucency at the distal tip. Eleven radiolucencies were > or =1 mm in width. A neutral stem orientation was significantly less likely to be associated with radiolucency (p = 0.026). The prevalence of radiolucent lines did not differ between patients managed with hemiarthroplasty and those managed with total shoulder arthroplasty, and it did not increase with longer periods of follow-up. CONCLUSIONS: In patients managed with shoulder arthroplasty, the fixation of a press-fit humeral component that has a tapered metaphyseal segment is comparable with that reported for cemented components and superior to that reported for press-fit cylindrical components. CLINICAL RELEVANCE: This type of fixation may provide an alternative to cementing of the humeral stem in individuals with glenohumeral osteoarthritis. PMID- 12571310 TI - Quantifying the extent of femoral head involvement in osteonecrosis. AB - BACKGROUND: There are numerous methods for quantifying the extent of osteonecrosis of the femoral head. However, there is no consensus regarding which method is the most reliable. The purpose of this study was to determine the reliability and prognostic accuracy of three commonly used methods for quantifying the extent of osteonecrosis of the femoral head. METHODS: Thirty-nine hips in twenty-five patients who had stage-I or II osteonecrosis of the femoral head, according to the grading system of the Association Research Circulation Osseous, were independently examined on two separate occasions by three observers of different specialty backgrounds and experience. Each observer used three methods to quantify the extent of osteonecrosis of the femoral head: (1) the index of necrotic extent, (2) the modified index of necrotic extent, and (3) the percentage of femoral head involvement. The interobserver and intraobserver agreement was determined for each method, and the ability of each method to predict the time to subchondral collapse was analyzed statistically. RESULTS: There was significantly valid agreement among the observers for all three methods (p < 0.001 for all three). The correlation coefficients demonstrated substantial agreement among raters when they measured the index of necrotic extent and the percent involvement and nearly perfect agreement when they measured the modified index of necrotic extent. Survivorship analysis revealed that the percent involvement (p < 0.05), index of necrotic extent (p < 0.007), and modified index of necrotic extent (p < 0.04) were prognostically significant predictors of subchondral fracture. CONCLUSIONS: Our results indicate that the index of necrotic extent, modified index of necrotic extent, and estimation of the percentage of involvement of the femoral head are reproducible and reliable methods for quantitatively evaluating the extent of osteonecrosis of the femoral head. We believe that the three methods can be utilized with confidence. Furthermore, they are clinically useful for identifying hips at greatest risk for subchondral collapse. PMID- 12571311 TI - Shoulder contracture and osseous deformity in obstetrical brachial plexus injuries. AB - BACKGROUND: The purposes of this study were to determine the prevalence of and the association between shoulder contracture and osseous deformity in a cohort of children with an obstetrical brachial plexus injury and to identify the risk factors for these complications. METHODS: In a retrospective cohort study, all children with an obstetrical brachial plexus injury who had been born between January 1991 and January 1998 at one academic medical center and all those with the same diagnosis who had been born elsewhere during the same period and were referred to the medical center within six weeks after delivery were evaluated at fixed time-intervals by one examiner. The patients underwent a final orthopaedic, neurological, and radiographic examination at a mean age of 3.7 years (range, one to seven years). Shoulder contracture was defined as a decrease in the passive range of motion in one or more directions compared with the range on the unaffected side. Osseous deformity was defined as a nonspherical humeral head or an abnormal glenoid. RESULTS: The prevalence of a shoulder contracture of >10 degrees was 56% (twenty-nine of fifty-two patients), and the prevalence of an osseous deformity was 33% (sixteen of forty-eight patients with complete radiographic follow-up). In the children in whom complete neurological recovery was delayed (i.e., recovery was more than three weeks after birth), the prevalence of shoulder contracture was 54% (thirteen of twenty-four patients) and the prevalence of osseous deformity was 26% (six of twenty-three patients). A strong association was noted between shoulder contracture and osseous deformity (p = 0.004). Directly after birth, the presence of a clavicular fracture was the only factor that was associated (p = 0.016) with the development of an osseous deformity (but not with a shoulder contracture). At a later stage, speed and extent of neurological recovery were related to shoulder contracture and osseous deformity. An asymmetric appearance was noticed in children who had a contracture, including those who had complete neurological recovery. CONCLUSIONS: The prevalence of shoulder contracture and osseous deformity in children with obstetrical brachial plexus injury was high, even in those with complete neurological recovery. These complications were strongly associated with one another. No symptom that appeared immediately after birth was identified as a factor that would predict the development of future shoulder contracture. A clavicular fracture was found to be significantly associated with the development of an osseous deformity at a later stage. PMID- 12571312 TI - Demineralization for inactivation of infectious retrovirus in systemically infected cortical bone: in vitro and in vivo experimental studies. AB - BACKGROUND: Clinical and experimental studies have demonstrated viral transmission through the transplantation of fresh-frozen infected bone. While sterilization methods sufficient to inactivate the human immunodeficiency virus (HIV) have been shown to markedly alter osteoconductive and osteoinductive properties of bone allografts, the ability of a process for creating demineralized bone matrix to abrogate transmission of a retrovirus has not been investigated, to our knowledge. We hypothesized that a clinically accepted demineralization procedure would alter the nucleic acids of the feline leukemia virus (FeLV, a retrovirus with a structure and replication cycle similar to those of HIV), inactivating the virus in infected bone and rendering it noninfectious. METHODS: Bone infected with FeLV was demineralized with a method employed for creating demineralized bone matrix powder. The effects of demineralization on cellular and (pro)viral nucleic acids were examined with use of gel electrophoresis and quantitative polymerase chain reaction, respectively. To compare the infectivity of the demineralized bone matrix with that of mineralized bone particles in cell cultures and in animals in which they had been implanted, we measured FeLV p27 antigen and (pro)viral nucleic acids as well as antiviral antibodies. RESULTS: Demineralization of FeLV-infected bone appeared to inactivate the virus by degradation and fragmentation of the DNA, rendering it noninfectious in both in vitro and in vivo test systems. In contrast, untreated mineralized FeLV-infected bone contained intact nucleic acids and readily transmitted the virus in both test systems. CONCLUSIONS: The demineralization process inactivated infectious retrovirus in infected cortical bone, thereby preventing disease transmission. PMID- 12571313 TI - Hypogastric artery disruption associated with acetabular fracture. A report of two cases. PMID- 12571314 TI - Intraoperative anaphylactic shock associated with bacitracin irrigation during revision total knee arthroplasty. A case report. PMID- 12571315 TI - Dumbbell-shaped iliopsoas bursitis penetrating the pelvic wall: a rare complication of hip arthrodesis. A case report. PMID- 12571316 TI - Traumatic posterior spondyloptosis at the lumbosacral junction. A case report. PMID- 12571318 TI - Subspecialization in orthopaedics. Has it been all for the better? PMID- 12571319 TI - The number and the scope of activity of orthopaedic clinician-scientists in the United States. PMID- 12571320 TI - Don't throw away the cane. 1956. PMID- 12571321 TI - Arthroscopic treatment of osteoarthritis of the knee. PMID- 12571322 TI - Head to head on spear tackler's spine: criteria and implications for return to play. PMID- 12571324 TI - Supplemental pedicle-screw implementation did not improve long-term functional outcomes after spinal arthrodesis. PMID- 12571323 TI - Differing approaches to the surgical correction of snapping hip. PMID- 12571326 TI - Hydroxyapatite cement was not as effective as intrafocal Kirschner-wire fixation for acute fractures of the distal part of the radius. PMID- 12571328 TI - Arthroscopic lavage or debridement did not reduce pain more than placebo did in patients with osteoarthritis. PMID- 12571331 TI - What's new in hand surgery. PMID- 12571332 TI - Addressing the crisis in US health care: moving beyond denial. PMID- 12571333 TI - Retrospective case record review: a blunt instrument that needs sharpening. PMID- 12571334 TI - International comparisons of the quality of health care: what do they tell us? PMID- 12571335 TI - Safe treatment or a shorter wait: Hobson's choice? PMID- 12571336 TI - Influence of attitudes and behaviour of GPs on prescribing costs. PMID- 12571338 TI - Can health care quality indicators be transferred between countries? AB - OBJECTIVE: To evaluate the transferability of primary care quality indicators by comparing indicators for common clinical problems developed using the same method in the UK and the USA. METHOD: Quality indicators developed in the USA for a range of common conditions using the RAND-UCLA appropriateness method were applied to 19 common primary care conditions in the UK. The US indicators for the selected conditions were used as a starting point, but the literature reviews were updated and panels of UK primary care practitioners were convened to develop quality indicators applicable to British general practice. RESULTS: Of 174 indicators covering 18 conditions in the US set for which a direct comparison could be made, 98 (56.3%) had indicators in the UK set which were exactly or nearly equivalent. Some of the differences may have related to differences in the process of developing the indicators, but many appeared to relate to differences in clinical practice or norms of professional behaviour in the two countries. There was a small but non-significant relationship between the strength of evidence for an indicator and the probability of it appearing in both sets of indicators. CONCLUSION: There are considerable benefits in using work from other settings in developing measures of quality of care. However, indicators cannot simply be transferred directly between countries without an intermediate process to allow for variation in professional culture or clinical practice. PMID- 12571339 TI - Views of older people on cataract surgery options: an assessment of preferences by conjoint analysis. AB - BACKGROUND: Key issues in the quality of care for people with cataracts in the UK include hospital waiting lists, complication rates from surgery, and the use of junior surgeons. The main objective of this study was to investigate the relative importance that older people attach to these factors when given theoretical choices over options for cataract surgery. METHOD: A systematic sample of 194 individuals aged 60-84 years on a general practice register in Nottingham were invited to take part in an interview based survey. Respondents ranked 11 "cataract surgery packages" containing different waiting list lengths, complication risks, and surgeon grades. Conjoint analysis was performed to determine the relative importance of these factors for individuals and for the group as a whole. RESULTS: Of the 194 subjects invited to participate, 146 (72%) completed the interview. For the group as a whole the "averaged importance" of the factors was: complication risk 45.8%; waiting time 41.1%, surgeon grade 13.1%. Analysis of importance scores for individuals showed that some were particularly concerned about complication risk while others were more concerned about waiting times. There was a strong negative correlation between importance scores for these factors (Spearman's rho -0.78, p<0.001). CONCLUSIONS: Most respondents thought that either risk of damage to sight and/or waiting time were important, while surgeon grade was relatively unimportant. The findings show that some potential cataract patients prefer a greater risk of complication combined with a short wait than a low complication rate and a longer wait. PMID- 12571340 TI - Development and validation of an international appraisal instrument for assessing the quality of clinical practice guidelines: the AGREE project. AB - BACKGROUND: International interest in clinical practice guidelines has never been greater but many published guidelines do not meet the basic quality requirements. There have been renewed calls for validated criteria to assess the quality of guidelines. OBJECTIVE: To develop and validate an international instrument for assessing the quality of the process and reporting of clinical practice guideline development. METHODS: The instrument was developed through a multi-staged process of item generation, selection and scaling, field testing, and refinement procedures. 100 guidelines selected from 11 participating countries were evaluated independently by 194 appraisers with the instrument. Following refinement the instrument was further field tested on three guidelines per country by a new set of 70 appraisers. RESULTS: The final version of the instrument contained 23 items grouped into six quality domains with a 4 point Likert scale to score each item (scope and purpose, stakeholder involvement, rigour of development, clarity and presentation, applicability, editorial independence). 95% of appraisers found the instrument useful for assessing guidelines. Reliability was acceptable for most domains (Cronbach's alpha 0.64 0.88). Guidelines produced as part of an established guideline programme had significantly higher scores on editorial independence and, after the publication of a national policy, had significantly higher quality scores on rigour of development (p<0.005). Guidelines with technical documentation had higher scores on that domain (p<0.0001). CONCLUSIONS: This is the first time an appraisal instrument for clinical practice guidelines has been developed and tested internationally. The instrument is sensitive to differences in important aspects of guidelines and can be used consistently and easily by a wide range of professionals from different backgrounds. The adoption of common standards should improve the consistency and quality of the reporting of guideline development worldwide and provide a framework to encourage international comparison of clinical practice guidelines. PMID- 12571341 TI - Are audits wasting resources by measuring the wrong things? A survey of methods used to select audit review criteria. AB - OBJECTIVES: This study measured the extent to which a systematic approach was used to select criteria for audit, and identified problems in using such an approach with potential solutions. DESIGN: A questionnaire survey using the Audit Criteria Questionnaire (ACQ), created, piloted, and validated for the purpose. Possible ACQ scores ranged from 0 to 1, indicating how systematically the criteria had been selected and how usable they were. SETTING: A stratified random sample of 10 audit leads in each of 83 randomly selected NHS trusts and all practices in each of 11 randomly selected primary care audit group areas in England and Wales. PARTICIPANTS: Audit leads of ongoing audits in each organisation in which a first data collection had started less than 12 months earlier and a second data collection was not completed. MAIN OUTCOME MEASURES: ACQ scores, problems identified in the audit criteria selection process, and solutions found. RESULTS: The mean ACQ score from all 83 NHS trusts and the 11 primary care audit groups was 0.52 (range 0.0-0.98). There was no difference between mean ACQ scores for criteria used in audits on clinical (0.51) and non clinical (0.52) topics. The mean ACQ scores from nationally organised audits (0.59, n=33) was higher than for regional (0.51, n=21), local (0.53, n=77), or individual organisation (0.52, n=335) audits. The mean ACQ score for published audit protocols (0.56) was higher than for locally developed audits (0.49). There was no difference in ACQ scores for audits reported by general practices (0.49, n=83) or NHS trusts (0.53, n=383). Problems in criteria selection included difficulties in coordination of staff to undertake the task, lack of evidence, poor access to literature, poor access to high quality data, lack of time, and lack of motivation. Potential solutions include investment in training, protected time, improved access to literature, support staff and availability of published protocols. CONCLUSIONS: Methods of selecting review criteria were often less systematic than is desirable. Published usable audit protocols providing evidence based review criteria with information on their provenance enable appropriate review criteria to be selected, so that changes in practice based on these criteria lead to real improvement in quality rather than merely change. The availability and use of high quality audit protocols would be a valuable contribution to the evolution of clinical governance. The ACQ should be developed into a tool to help in selecting appropriate criteria to increase the effectiveness of audit. PMID- 12571342 TI - Attitudes and behaviour of general practitioners and their prescribing costs: a national cross sectional survey. AB - BACKGROUND: General practitioner (GP) prescribing accounts for about 10% of NHS expenditure. GPs at the top of the range have annual prescribing costs that are almost twice as much as those at the bottom of the range. This variation cannot be accounted for purely in terms of differences in underlying need for health care. OBJECTIVES: To describe the relationship between GPs' prescribing costs and their attitudes towards prescribing decisions and prescribing information sources, and to identify potentially modifiable attitudinal and behavioural factors associated with high cost prescribing. DESIGN: A postal questionnaire was designed on the basis of hypotheses developed from a literature search and an earlier qualitative survey. This questionnaire was sent to a national sample of GPs with equal numbers of practices in the upper, middle, and lowest quintile of prescribing costs. SETTING: GP practices in England. PARTICIPANTS: 1714 GPs in NHS practice. OUTCOME MEASURES: GPs' self-reported practices, attitudes and personal characteristics. RESULTS: There was a 64% response rate. Responders were more likely to be from larger practices, in less deprived areas, and with lower prescribing costs than were non-responders. Multivariable analysis showed that GPs with high prescribing costs were significantly more likely to work in dispensing practices, in practices with low income populations, in single handed practices, and in practices without a GP trainer. They were also significantly more likely to see drug company representatives more frequently, to prescribe newly available drugs more freely, to prescribe more readily to patients who expect a prescription, to report high levels of frustration from lack of time in the consultation, to find unsatisfactory those consultations which ended in advice only, and to express dissatisfaction with their review methods for repeat prescribing. They were significantly less likely to find useful criticism of prescribing habits by colleagues, and to check the BNF rather than other sources when uncertain about an aspect of drug treatment. CONCLUSIONS: While they cannot be held to have a causal relationship, the pattern of attitudes towards prescribing of GPs in the highest quintile of prescribing costs provide the basis for developing an educational intervention which may be an acceptable method of modifying the attitudes of GPs and consequently reducing their prescribing costs. PMID- 12571343 TI - From aviation to medicine: applying concepts of aviation safety to risk management in ambulatory care. AB - The development of a medical risk management programme based on the aviation safety approach and its implementation in a large ambulatory healthcare organisation is described. The following key safety principles were applied: (1). errors inevitably occur and usually derive from faulty system design, not from negligence; (2). accident prevention should be an ongoing process based on open and full reporting; (3). major accidents are only the "tip of the iceberg" of processes that indicate possibilities for organisational learning. Reporting physicians were granted immunity, which encouraged open reporting of errors. A telephone "hotline" served the medical staff for direct reporting and receipt of emotional support and medical guidance. Any adverse event which had learning potential was debriefed, while focusing on the human cause of error within a systemic context. Specific recommendations were formulated to rectify processes conducive to error when failures were identified. During the first 5 years of implementation, the aviation safety concept and tools were successfully adapted to ambulatory care, fostering a culture of greater concern for patient safety through risk management while providing support to the medical staff. PMID- 12571344 TI - Process evaluation on quality improvement interventions. AB - To design potentially successful quality improvement (QI) interventions, it is crucial to make use of detailed breakdowns of the implementation processes of successful and unsuccessful interventions. Process evaluation can throw light on the mechanisms responsible for the result obtained in the intervention group. It enables researchers and implementers to (1). describe the intervention in detail, (2). check actual exposure to the intervention, and (3). describe the experience of those exposed. This paper presents a framework containing features of QI interventions that might influence success. Attention is paid to features of the target group, the implementers or change agents, the frequency of intervention activities, and features of the information imparted. The framework can be used as a starting point to address all three aspects of process evaluation mentioned above. Process evaluation can be applied to small scale improvement projects, controlled QI studies, and large scale QI programmes; in each case it plays a different role. PMID- 12571345 TI - Research designs for studies evaluating the effectiveness of change and improvement strategies. AB - The methods of evaluating change and improvement strategies are not well described. The design and conduct of a range of experimental and non-experimental quantitative designs are considered. Such study designs should usually be used in a context where they build on appropriate theoretical, qualitative and modelling work, particularly in the development of appropriate interventions. A range of experimental designs are discussed including single and multiple arm randomised controlled trials and the use of more complex factorial and block designs. The impact of randomisation at both group and individual levels and three non experimental designs (uncontrolled before and after, controlled before and after, and time series analysis) are also considered. The design chosen will reflect both the needs (and resources) in any particular circumstances and also the purpose of the evaluation. The general principle underlying the choice of evaluative design is, however, simple-those conducting such evaluations should use the most robust design possible to minimise bias and maximise generalisability. PMID- 12571346 TI - Healthcare redesign: meaning, origins and application. AB - Healthcare organisations are using redesign to tackle variation in the quality of care and improve public satisfaction. It is represented as a radical challenge to traditional assumptions and practices which involves thinking through the best process to achieve speedy and effective patient care, identifying delays, unnecessary steps, or potential for error, and redesigning the process to improve the quality of care. This paper explores the meaning of redesign using practical illustrations. It examines its theoretical origins, particularly total quality management (TQM)/continuous quality improvement (CQI) and re-engineering, and assesses evidence which may inform its application. This evidence suggests that clinical ownership and senior management support will be essential. Redesign seeks to balance the more gradual approach of TQM with the organisation-wide lateral thinking of re-engineering. An incremental negotiated approach seems more likely to ensure clinical ownership, but carries a risk that QI will remain small scale with little impact on the wider organisation. Inclusion of some re engineering techniques may help to overcome this difficulty. Evidence suggests that most QI techniques achieve only partial success. This may pose difficulties for redesign, which has generated high political expectations that it can solve long term problems in health care. PMID- 12571347 TI - The hazards of hospitalization. 1964. PMID- 12571349 TI - The prevention and treatment of childhood obesity. AB - The effectiveness of interventions used in the prevention and treatment of childhood obesity published in a recent issue of Effective Heath Care is reviewed. PMID- 12571352 TI - Drosophila DEG/ENaC pickpocket genes are expressed in the tracheal system, where they may be involved in liquid clearance. AB - The Drosophila tracheal system and mammalian airways are branching networks of tubular epithelia that deliver oxygen to the organism. In mammals, the epithelial Na(+) channel (ENaC) helps clear liquid from airways at the time of birth and removes liquid from the airspaces in adults. We tested the hypothesis that related Drosophila degenerin (DEG)/ENaC family members might play a similar role in the fly. Among 16 Drosophila DEG/ENaC genes, called pickpocket (PPK) genes, we found 9 expressed in the tracheal system. By in situ hybridization, expression appeared in late-stage embryos after tracheal tube formation, with individual PPK genes showing distinct temporal and spatial expression patterns as development progressed. Promoters for several PPK genes drove reporter gene expression in the larval and adult tracheal systems. Adding the DEG/ENaC channel blocker amiloride to the medium inhibited liquid clearance from the trachea of first instar larvae. Moreover, when RNA interference was used to silence PPK4 and PPK11, larvae failed to clear tracheal liquid. These data suggest substantial molecular diversity of DEG/ENaC channel expression in the Drosophila tracheal system where the PPK proteins likely play a role in Na(+) absorption. Extensive similarities between Drosophila and mammalian airways offer opportunities for genetic studies that may decipher further the structure and function of DEG/ENaC proteins and development of the airways. PMID- 12571353 TI - Packing helices in proteins by global optimization of a potential energy function. AB - An efficient method has been developed for packing alpha-helices in proteins. It treats alpha-helices as rigid bodies and uses a simplified Lennard-Jones potential with Miyazawa-Jernigan contact-energy parameters to describe the interactions between the alpha-helical elements in this coarse-grained system. Global conformational searches to generate packing arrangements rapidly are carried out with a Monte Carlo-with-minimization type of approach. The results for 42 proteins show that the approach reproduces native-like folds of alpha helical proteins as low-energy local minima of this highly simplified potential function. PMID- 12571354 TI - Variation in gene expression patterns in follicular lymphoma and the response to rituximab. AB - Analysis of the patterns of gene expression in follicular lymphomas from 24 patients suggested that two groups of tumors might be distinguished. All patients, whose biopsies were obtained before any treatment, were treated with rituximab, a monoclonal antibody directed against the B cell antigen, CD20. Gene expression patterns in the tumors that subsequently failed to respond to rituximab appeared more similar to those of normal lymphoid tissues than to gene expression patterns of tumors from rituximab responders. These findings suggest the possibility that the response of follicular lymphoma to rituximab treatment may be predicted from the gene expression pattern of tumors. PMID- 12571355 TI - Long-term regional suppression of pink bollworm by Bacillus thuringiensis cotton. AB - Despite the potentially profound impact of genetically modified crops on agriculture and the environment, we know little about their long-term effects. Transgenic crops that produce toxins from Bacillus thuringiensis (Bt) to control insects are grown widely, but rapid evolution of resistance by pests could nullify their benefits. Here, we present theoretical analyses showing that long term suppression of pest populations is governed by interactions among reproductive rate, dispersal propensity, and regional abundance of a Bt crop. Supporting this theory, a 10-year study in 15 regions across Arizona shows that Bt cotton suppressed a major pest, pink bollworm (Pectinophora gossypiella), independent of demographic effects of weather and variation among regions. Pink bollworm population density declined only in regions where Bt cotton was abundant. Such long-term suppression has not been observed with insecticide sprays, showing that transgenic crops open new avenues for pest control. The debate about putative benefits of Bt crops has focused primarily on short-term decreases in insecticide use. The present findings suggest that long-term regional pest suppression after deployment of Bt crops may also contribute to reducing the need for insecticide sprays. PMID- 12571356 TI - Cell transfection in vitro and in vivo with nontoxic TAT peptide-liposome-DNA complexes. AB - Liposomes modified with TAT peptide (TATp-liposomes) showed fast and efficient translocation into the cell cytoplasm with subsequent migration into the perinuclear zone. TATp-liposomes containing a small quantity (4 weeks is associated with aggression toward intruders and a down-regulation of brain allopregnanolone (Allo) content. SI of female mice fails to down-regulate brain Allo content or to induce aggressiveness. Fluoxetine (Prozac in clinical use) is an S- and R-fluoxetine (FLX) mixture, which in mammals is metabolized into S- and R-norfluoxetine (NFLX). The S isomers of FLX and NFLX are more active than their respective R isomers in normalizing brain Allo down-regulation and in reducing the aggressiveness induced by SI. Thus, FLX stereospecifically reduces brain Allo down-regulation and the aggressiveness induced by SI, whereas serotonin (5-HT) uptake inhibition lacks stereospecificity. The doses of S-FLX and S-NFLX that reduce aggressiveness and Allo brain content down-regulation induced by SI are at least one order of magnitude lower than the doses that block 5-HT reuptake. Doses of imipramine that inhibit 5-HT uptake neither reduce aggressiveness nor normalize brain Allo down-regulation. We conclude that Allo brain content normalization is a better candidate than 5-HT reuptake inhibition to explain the reduction of aggressiveness elicited by S-FLX and S-NFLX. PMID- 12571362 TI - Deletion of the p66Shc longevity gene reduces systemic and tissue oxidative stress, vascular cell apoptosis, and early atherogenesis in mice fed a high-fat diet. AB - Several experimental and clinical studies have shown that oxidized low-density lipoprotein and oxidation-sensitive mechanisms are central in the pathogenesis of vascular dysfunction and atherogenesis. Here, we have used p66(Shc-/-) and WT mice to investigate the effects of high-fat diet on both systemic and tissue oxidative stress and the development of early vascular lesions. To date, the p66(Shc-/-) mouse is the unique genetic model of increased resistance to oxidative stress and prolonged life span in mammals. Computer-assisted image analysis revealed that chronic 21% high-fat treatment increased the aortic cumulative early lesion area by approximately 21% in WT mice and only by 3% in p66(Shc-/-) mice. Early lesions from p66(Shc-/-) mice had less content of macrophage-derived foam cells and apoptotic vascular cells, in comparison to the WT. Furthermore, in p66(Shc-/-) mice, but not WT mice, we found a significant reduction of systemic and tissue oxidative stress (assessed by isoprostanes, plasma low-density lipoprotein oxidizability, and the formation of arterial oxidation-specific epitopes). These results support the concept that p66(Shc-/-) may play a pivotal role in controlling systemic oxidative stress and vascular diseases. Therefore, p66(Shc) might represent a molecular target for therapies against vascular diseases. PMID- 12571363 TI - Reduced glucose tolerance is associated with poor memory performance and hippocampal atrophy among normal elderly. AB - Poor glucose tolerance and memory deficits, short of dementia, often accompanies aging. The purpose of this study was to ascertain whether, among nondiabetic, nondemented middle-aged and elderly individuals, poorer glucose tolerance is associated with reductions in memory performance and smaller hippocampal volumes. We studied 30 subjects who were evaluated consecutively in an outpatient research setting. The composition of the participant group was 57% female and 68.6 +/- 7.5 years of age; the participants had an average education of 16.2 +/- 2.3 years, a score on the Mini Mental State Examination of 28.6 +/- 1.5, a glycosylated hemoglobin (HbA1C) of 5.88 +/- 0.74%, and a body mass index of 24.9 +/- 4.1 kg/m(2). Glucose tolerance was measured by an i.v. glucose tolerance test. Memory was tested by using the Wechsler Paragraphs recall tests at the time of administering the i.v. glucose tolerance test. The hippocampus and other brain volumes were measured by using validated methods on standardized MRIs. Decreased peripheral glucose regulation was associated with decreased general cognitive performance, memory impairments, and atrophy of the hippocampus, a brain area that is key for learning and memory. These associations were independent of age and Mini Mental State Examination scores. Therefore, these data suggest that metabolic substrate delivery may influence hippocampal structure and function. This observation may bring to light a mechanism for aging brain injury that may have substantial medical impact, given the large number of elderly individuals with impaired glucose metabolism. PMID- 12571364 TI - Glycosaminoglycan binding and oligomerization are essential for the in vivo activity of certain chemokines. AB - During organogenesis, immunosurveillance, and inflammation, chemokines selectively recruit leukocytes by activating seven-transmembrane-spanning receptors. It has been suggested that an important component of this process is the formation of a haptotactic gradient by immobilization of chemokines on cell surface glycosaminoglycans (GAGs). However, this hypothesis has not been experimentally demonstrated in vivo. In the present study we investigated the effect of mutations in the GAG binding sites of three chemokines, monocyte chemoattractant protein-1/CC chemokine ligand (CCL)2, macrophage-inflammatory protein-1beta/CCL4, and RANTES/CCL5, on their ability to recruit cells in vivo. These mutant chemokines retain chemotactic activity in vitro, but they are unable to recruit cells when administered intraperitoneally. Additionally, monomeric variants, although fully active in vitro, are devoid of activity in vivo. These data demonstrate that both GAG binding and the ability to form higher-order oligomers are essential for the activity of particular chemokines in vivo, although they are not required for receptor activation in vitro. Thus, quaternary structure of chemokines and their interaction with GAGs may significantly contribute to the localization of leukocytes beyond migration patterns defined by chemokine receptor interactions. PMID- 12571365 TI - STAT3 deletion during hematopoiesis causes Crohn's disease-like pathogenesis and lethality: a critical role of STAT3 in innate immunity. AB - Signal transducer and activator of transcription 3 (STAT3) is a key transcriptional mediator for many cytokines and is essential for normal embryonic development. We have generated a unique strain of mice with tissue-specific disruption of STAT3 in bone marrow cells during hematopoiesis. This specific STAT3 deletion causes death of these mice within 4-6 weeks after birth with Crohn's disease-like pathogenesis in both the small and large intestine, including segmental inflammatory cell infiltration, ulceration, bowel wall thickening, and granuloma formation. Deletion of STAT3 causes significantly increased cell autonomous proliferation of cells of the myeloid lineage, both in vivo and in vitro. Most importantly, Stat3 deletion during hematopoiesis causes overly pseudoactivated innate immune responses. Although inflammatory cytokines, including tumor necrosis factor alpha and IFN-gamma, are overly produced in these mice, the NAPDH oxidase activity, which is involved in antimicrobial and innate immune responses, is inhibited. The signaling responses to lipopolysaccharide are changed in the absence of STAT3, leading to enhanced NF-kappa B activation. Our results suggest a model in which STAT3 has critical roles in the development and regulation of innate immunity, and deletion of STAT3 during hematopoiesis results in abnormalities in myeloid cells and causes Crohn's disease-like pathogenesis. PMID- 12571366 TI - Primary microcephaly: microcephalin and ASPM determine the size of the human brain. PMID- 12571367 TI - Sex and the single gene: meiotic silencing by unpaired DNA. PMID- 12571368 TI - Reflective consciousness and the emergence of memes: serial evolutionary pathways? PMID- 12571369 TI - A metaphysics of living systems: the Yoga-Vasistha view. AB - The Yoga-Vasistha is a rich and complex philosophical 'poem' (kavya) of epic length, written in classical Sanskrit by an unknown author some time between the 6th and 13th centuries CE, probably around the 7th century. It is notable for its eloquent praise of self-effort and enquiry or analysis, and for its severe disparagement of the notion of fate. It views consciousness as (a) characterizing all living forms (including plant and insect life), (b) being atomic, and (c) analogous to the emergence of waves and whirlpools in water; it therefore grapples with what today would be called the problems of reductionism and emergentism. Notions of the survival of the fittest, and of a dynamic process of creation and loss, are expressed with characteristic force. The paper presents a selection of verses (in an English translation) setting forth these views, and a brief analysis of their implications. PMID- 12571370 TI - A passion for plant life. PMID- 12571371 TI - Monoclonal antibody affinity purification of a 78 kDa membrane protein of Leishmania donovani of Indian origin and its role in host-parasite interaction. AB - Monoclonal antibodies were raised against pathogenic promastigotes of Leishmania donovani of Indian origin. Among these, one was used for immuno-affinity purification of a 78 kDa membrane protein present in both the amastigote and promastigote forms of the parasite. Results of immunoblot experiments with the anti-78 kDa antibody revealed that the protein was present only in parasites belonging to the L. donovani complex. The expression of the protein was observed to be the same during different phases of growth of the promastigotes. Therefore, the 78 kDa protein is neither stage-specific nor differentially regulated. Surface iodination and subcellular fractionation of the promastigotes indicated that the protein was localized on the cell surface. The 78 kDa protein was found to inhibit the binding of promastigotes to macrophages significantly, suggesting that it may play a role in the process of infection. Thus, here we report the purification of a surface protein of L. donovani of Indian origin, which may play an important role in the process of infection. PMID- 12571372 TI - Alterations in tumour suppressor gene p53 in human gliomas from Indian patients. AB - Alterations in tumour suppressor p53 gene are the most common defects seen in a variety of human cancers. In order to study the significance of the p53 gene in the genesis and development of human glioma from Indian patients, we checked 44 untreated primary gliomas for mutations in exons 5-9 of the p53 gene by PCR-SSCP and DNA sequencing. Sequencing analysis revealed six missense mutations. The incidence of p53 mutations was 13.6% (6 of 44). All the six mutations were found to be located in the central core domain of p53, which carries the sequence specific DNA-binding domain. These results suggest a rather low incidence but a definite involvement of p53 mutations in the gliomas of Indian patients. PMID- 12571373 TI - Characterization of urinary volatiles in Swiss male mice (Mus musculus): bioassay of identified compounds. AB - The present study was carried out to investigate the chemical nature of the urine of male mice and to assess its bioactivity. Urine of mature male mice was extracted with dichloromethane (1 : 1 ratio v/v) and analysed by gas chromatography linked mass-spectrometry (GC-MS). Ten different compounds such as alkanes, alcohols, etc. were detected in the urine. Among the ten, five compounds are specific to males namely, 3-cyclohexene-1-methanol (I), 3-amino-s-triazole (II), 4-ethyl phenol (III), 3-ethyl-2,7-dimethyl octane (IV) and 1-iodoundecane (V). The compound, 4-ethylphenol, has been previously reported in several strains of male mice. Furthermore, the compounds (II) and (IV) are closely similar to 2 sec-butylthiazole and dehydro-exo-brevicomin compounds which have already been reported in male mice. Bioassay revealed that compounds (II), (III) and (IV) were responsible for attracting females and in inducing aggression towards males, as compared to the other compounds i.e. (I) and (V). The results indicate that these three volatiles (II, III and IV) of male mice appear to act as attractant of the opposite sex. PMID- 12571374 TI - Induction of blindness by formoguanamine hydrochloride in adult male roseringed parakeets (Psittacula krameri). AB - Formoguanamine (2,4-diamino-s-triazine) was known to be an effective chemical agent in inducing blindness in poultry chicks, but not in adult birds. The present study was undertaken to demonstrate the influences, if any, of this chemical on the visual performance and retinal histology in an adult sub-tropical wild bird the roseringed parakeet (Psittacula krameri). Formoguanamine (FG) hydrochloride was subcutaneously injected into adult parakeets at the dosage of 25 mg (dissolved in 0.75 ml physiological saline)/100 g body weight/day, for two consecutive days while the control birds were injected only with the placebo. The effects were studied after 10, 20, and 30 days of the last treatment of FG. Within 24 h of the treatment of FG, about 90% of the total birds exhibited lack of visual responses to any light stimulus and even absence of pupillary light reactions. The remaining birds became totally blind on the day following the last injection of FG and remained so till the end of investigation. At the microscopic level, conspicuous degenerative changes were noted in the outer pigmented epithelium and the photoreceptive layer of rods and cones in the retinas of FG treated birds. A significant reduction in the thickness of the outer nuclear layer was also found in the retinas of FG treated parakeets, compared to that in the control birds. However, the inner cell layers of the retina in the control and FG administered parakeets were almost identical. It deserves special mention that the effects of FG, noted after 30 days of last treatment, were not very different from those noted just after 10 days of treatment. Collectively, the results of the present investigation demonstrate that FG can be used as a potent pharmacological agent for inducing irreversible blindness through selective damage in retinal tissue even in the adult wild bird, thereby making FG treatment an alternative euthanasic device to a cumbersome, stressful, surgical method of enucleation of the ocular system for laboratory studies. PMID- 12571375 TI - Postnatal growth, age estimation and development of foraging behaviour in the fulvous fruit bat Rousettus leschenaulti. AB - This study documents the postnatal growth, age estimation and development of the foraging behaviour of the fulvous fruit bat Rousettus leschenaulti under captive conditions. At birth, the young were naked and pink with closed eyes and folded pinnae. By day four of age, their eyes had opened and the pups began to move. The mean length of forearm in 5-day-old pups was 24.9 mm and body mass was 10.8 g, equivalent to 32.3% and 14.2% of the values from postpartum females. The length of forearm and body mass increased linearly until 45 and 50 days, respectively, and thereafter maintained an apparent stability. The epiphyseal gap of the fourth metacarpal-phalangeal joint increased until 15 days, then decreased linearly until 75 days and thereafter closed. Age was estimated quantitatively, based on linear changes observed in the length of the forearm and epiphyseal gap. Pups began to roost separately, but adjacent to their mothers when 30 days old and flew clumsily when they were about 40 days old. After attaining clumsy flight, the young bats made independent foraging attempts feebly by biting and licking small fruit pieces. Young bats were engaged in suckling as well as ingesting fruits when they were about 50 days old. Between 55 and 65 days, they flew well and fed on fruits. At the age of 75 days, the young bats were completely weaned and at two months, their foraging behaviour was similar to that of their mothers. There was no significant difference in the growth pattern of the young maintained in captivity compared with those under natural conditions. PMID- 12571377 TI - Reliability of the ACTH low dose test in the evaluation of adrenal insufficiency. PMID- 12571376 TI - Applications of inulin and oligofructose in health and nutrition. AB - Inulin and oligofructose belong to a class of carbohydrates known as fructans. The main sources of inulin and oligofructose that are used in the food industry are chicory and Jerusalem artichoke. Inulin and oligofructose are considered as functional food ingredients since they affect the physiological and biochemical processes in rats and human beings, resulting in better health and reduction in the risk of many diseases. Experimental studies have shown their use as bifidogenic agents, stimulating the immune system of the body, decreasing the pathogenic bacteria in the intestine, relieving constipation, decreasing the risk of osteoporosis by increasing mineral absorption, especially of calcium, reducing the risk of atherosclerosis by lowering the synthesis of triglycerides and fatty acids in the liver and decreasing their level in serum. These fructans modulate the hormonal level of insulin and glucagon, thereby regulating carbohydrate and lipid metabolism by lowering the blood glucose levels; they are also effective in lowering the blood urea and uric acid levels, thereby maintaining the nitrogen balance. Inulin and oligofructose also reduce the incidence of colon cancer. The biochemical basis of these beneficial effects of inulin and oligofructose have been discussed. Oligofructose are non cariogenic as they are not used by Streptococcus mutans to form acids and insoluble glucans that are the main culprits in dental caries. Because of the large number of health promoting functions of inulin and oligofructose, these have wide applications in various types of foods like confectionery, fruit preparations, milk desserts, yogurt and fresh cheese, baked goods, chocolate, ice cream and sauces. Inulin can also be used for the preparation of fructose syrups. PMID- 12571378 TI - A hypothesis of epiarachnoidal growth of vestibular schwannoma at the cerebello pontine angle: surgical importance. AB - AIMS: The purpose of this study is to clarify the rearrangement of the arachnoid membrane on the vestibular schwannoma during its growth in relation to adjacent neurovascular structures for a better understanding of dissecting plane of arachnoid during surgery. METHODS: Arachnoid membrane over the tumour was investigated during surgery with suboccipital transmeatal approach in twenty-six tumours. All microsurgical procedures were recorded with a video and reviewed. The tumour growth was classified into five stages depending upon the tumour diameter in the cerebello-pontine (CP) angle: Stage 1; purely intracanalicular (2 cases), Stage 2; less than 5 mm (2 cases), Stage 3; > or = 5 and <15 mm (8 cases), Stage 4; > or = 15 and <25 mm (9 cases) and Stage 5; > or = 25 mm (5 cases). Rearrangement of the arachnoid on the tumour was conceptualised throughout all stages. RESULTS: All tumours of Stage 1 and 2 were entirely located in the subarachnoid space of the cerebello-pontine cistern without arachnoidal rearrangement, while all tumours of Stages 3 to 5 were enveloped, in the CP angle, with invaginated arachnoid membrane consisting of cerebello-pontine cistern except two surfaces; the medial pole and the tumour surface under the facial and cochlear nerves near the porus. CONCLUSION: The tumour originates subarachnoidally within the internal auditory meatus (IAM) and grows epiarachnoidally in the CP angle. Rearrangement of the arachnoid begins with its adhesion on the medial pole of the tumour along the porus, resulting in the arachnoidal invagination into the cerebello-pontine cistern with further growing of the tumour. PMID- 12571379 TI - Effect of 12 months of recombinant human growth hormone replacement therapy on insulin sensitivity in GH-deficient adults as determined by different methods. AB - BACKGROUND: Controversial results have been obtained in measuring insulin sensitivity (S(I)) during recombinant human growth hormone (rhGH) treatment in adult growth hormone deficient (GH-deficient) patients. AIMS: The aim of our study was to estimate S(I) before and during treatment using three different methods for quantifying insulin sensitivity in GH-deficient adults treated with rhGH. SETTINGS AND DESIGN: Twenty-one GH-deficient adults were treated with rhGH during 12 months. S(I) was estimated using Minimal model analysis, Homeostatic Model of Assessment (HOMA) and Quantitative Insulin Sensitivity Check Index (QUICKI) before and after 3, 6, 9 and 12 months of rhGH therapy. MATERIAL AND METHODS: Oral Glucose Tolerance Test (OGTT) and Frequently Sampled Intravenous Glucose Tolerance Test (FSIGT) were performed in each patient at respective time intervals. QUICKI and HOMA were calculated using basal values of glucose and insulin from FSIGT. Minimal model computer analysis was calculated from glucose and insulin data obtained during FSIGT. STATISTICAL ANALYSIS: Area under the curve for glucose, insulin and C-peptide were calculated using trapezoidal rule from OGTT data. Differences and correlations were tested using ANOVA for repeated measures, Wilcoxon's matched-paired test, paired t-test, Pearson's correlation and Bland Altman plot. RESULTS: There were no significant changes in S(I) using Minimal model analysis and QUICKI during rhGH treatment. On the contrary, HOMA analysis indicated significant deterioration in S(I) after 12 months of therapy. CONCLUSION: Our study did not demonstrate any changes in S(I) using Minimal model and QUICKI analysis, while there was significant increase in insulin resistance using HOMA model. We suggest that the choice of method for the determination of S(I) may influence the interpretation of results concerning the effect of rhGH therapy on S(I) in GH-deficient adults. PMID- 12571380 TI - The role of ACE gene polymorphism in rapidity of progression of focal segmental glomerulosclerosis. AB - BACKGROUND: The insertion/deletion (I/D) polymorphism of angiotensin converting enzyme (ACE) gene has been associated with progression of renal diseases. AIMS: We investigated its role in the rate of progression of focal segmental glomerulosclerosis (FSGS). METHODS: Forty-seven patients with end-stage renal disease (ESRD) due to FSGS were evaluated. RESULTS: The distribution of ACE genotype was II-25.5%, ID-55.5%, and DD-19%, as compared to 40 controls with genotype of 7.5%, 60%, and 32.5%, respectively (p= NS). In African Americans (AA) the gene frequencies among patients and controls were I-43%, D-57% vs I-36%, D 64%, respectively. This was different than the gene frequencies in White/Hispanic (W/H) patients I-61.5%, D-38.5% vs I-38.6%, D-61.4%, in controls (P < 0.05). In 22 patients with rapid progression (RP) of FSGS to ESRD the genotype distribution was II-18%, ID -64%, and DD-18%. In 25 patients with FSGS who progressed slowly (SP) the genotype was similar (II-32%, ID-48% and DD-20%, P >0.05). With respect to rate of progression, D allele frequency was similar in AA patients (RP 64% vs SP 50%) and W/H patients (RP 36% vs SP 40%). CONCLUSION: Our study reveals no association between the I/D polymorphism of the ACE gene and the presence of and rapidity progression of FSGS. PMID- 12571381 TI - Magnetic resonance arthrography in recurrent anterior shoulder instability as compared to arthroscopy: a prospective comparative study. AB - AIM: To evaluate the accuracy of magnetic resonance (MR) arthrographic imaging in the diagnosis of glenoid labral and ligament tears in recurrent shoulder instability. SETTINGS AND DESIGN: Prospective, comparative study at a tertiary care centre. MATERIAL AND METHODS: Patients with three or more episodes of anterior shoulder dislocation were enrolled in the study. They were subjected to magnetic resonance arthrography (MRA) for delineation of abnormalities. The findings obtained at MRA were compared with those found at arthroscopy and surgical exploration. RESULTS: MRA detected glenoid tears in all 22 patients with 20 (90%) patients having antero-inferior tears, 3 (14%) patients had superior labral involvement and 2 (10%) patients had posterior labral abnormality. On arthroscopy, antero-inferior, superior and posterior labral tear were found in 21 (95%), 5 (22%) and 7 (32%) patients respectively. MRA showed a sensitivity of 95%, and a specificity of 100% for the detection of the antero-inferior labral tears. The sensitivity of MRA for the detection of superior, middle and inferior glenohumeral ligament tear was 83%, 80% and 86% with a specificity of 100%, 71% and 93% respectively. MRA was 100% sensitive for the detection of rotator cuff injuries and detection of bony lesions like Hill-Sach's and bony Bankart's lesion. CONCLUSIONS: MRA is a sensitive and specific modality for evaluation of anterior shoulder instability. PMID- 12571382 TI - Neurological manifestations of snake bite in Sri Lanka. AB - BACKGROUND AND AIMS: Snake bite is an important cause of mortality and morbidity in certain parts of Sri Lanka. This study was designed to determine the offending snakes, neurological manifestations, disease course, and outcome in neurotoxic envenomation. METHODS AND MATERIAL: Fifty six consecutive patients admitted with neurological manifestations following snake bite were studied prospectively. Data were obtained regarding the offending snakes, neurological symptoms, time taken for onset of symptoms, neurological signs, and time taken for recovery. RESULTS: The offending snake was Russell's viper in 27(48.2%), common and Sri Lankan krait in 19(33.9%), cobra in 3(5.4%), and unidentified in 7(12.5%). Ptosis was the commonest neurological manifestation seen in 48(85.7%) followed by ophthalmoplegia (75%), limb weakness (26.8%), respiratory failure (17.9%), palatal weakness (10.7%), neck muscle weakness (7.1%), and delayed sensory neuropathy (1.8%). Neurological symptoms were experienced usually within 6 hours after the bite. Following administration of antivenom, the signs of recovery became evident within a few hours to several days. The duration for complete recovery ranged from four hours to two weeks. CONCLUSIONS: Complete recovery of neuromuscular weakness was observed in all patients except for one who died with intracerebral haemorrhage shortly after admission. PMID- 12571383 TI - Evaluation of low dose ACTH stimulation test in suspected secondary adrenocortical insufficiency. AB - BACKGROUND: Several studies in the last few years have shown that the standard 250 micro g dose used in ACTH stimulation test may be very high and have suggested that a dose of 1 micro g may be sufficient for evaluating hypothalamo pituitary adrenal (HPA) axis. AIMS: To evaluate the role of low dose ACTH stimulation test in patients with suspected Secondary Adrenal Insufficiency (SAI). SUBJECTS AND METHODS: Thirty-one patients of suspected SAI with a documented pituitary lesion and seven normal healthy controls were included in the study. All the subjects underwent ACTH stimulation test with standard high dose (HD= 250 microg) and low dose (LD= 1 microg) ACTH. Insulin Induced Hypoglycaemia (IIH) test was done in 7 out of 9 patients in whom the results of the two tests were discordant. The cut off for normal HD stimulation test was taken as peak cortisol response > 18 microg/dl and for LD test, either a cortisol response of > 18 microg/dl or an increment of more than 7 micro g/dl over the basal value at any time during the test, on the basis of response observed in controls. RESULTS: 22/31 patients (74%) in SAI group showed concordant results with both tests, whereas nine patients had discordant results. These nine patients showed AI with LD ACTH, but HD ACTH test showed a normal response. In 7 of these 9 patients who underwent IIH, AI was confirmed in 6. CONCLUSION: The LD ACTH stimulation test seems to be better than HD ACTH stimulation test for evaluating HPA axis in patients with suspected SAI. When basal cortisol is normal, LD ACTH stimulation test detects subtle SAI. PMID- 12571384 TI - Orofacial pain and numb chin syndrome as the presenting symptoms of a metastatic prostate cancer. AB - We describe a patient with orofacial pain as the presenting symptom caused by a mandibular metastasis from a previously undiagnosed cancer of the prostate. This possibility should be considered in the differential diagnosis of male patients presenting with orofacial pain. PMID- 12571385 TI - A unique presentation of retroclival chordoma. AB - Chordomas are rare tumours which arise from remnants of the primitive notochord. They occur primarily in the sacrum, clivus and cervical regions. We report a case of retroclival chordoma which presented as an extradural haemorrhage following minor trauma. The underlying tumour was not apparent on imaging performed immediately following the event, and chordoma presenting in this manner has not previously been described in the literature. The tumour became apparent on subsequent imaging, and progressed despite surgical debulking and radiotherapy. PMID- 12571386 TI - Spontaneous rupture of a left gastroepiploic artery aneurysm. AB - Gastroepiploic aneurysms are extremely rare. They occur mainly in elderly men and in 90% of cases are ruptured at presentation. Visceral aneurysms though rare should be borne in mind in cases of unexplained haemorrhagic shock. We present a case of a 79-year-old man who presented with abdominal pain, hypotension and anaemia but no obvious source of bleeding. He had undergone a prior aorto bifemoral graft. The patient refused an operation and died the following day. PMID- 12571387 TI - Electro convulsive therapy in a pre-pubertal child with severe depression. AB - Electro Convulsive Therapy (ECT) in pre-pubertal children is a controversial and underreported treatment. Even though the effectiveness and side effects of ECT in adolescents are comparable with those in adults, there is a pervasive reluctance to use ECT in children and adolescents. We report the case of a pre-pubertal child in an episode of severe depression with catatonic features, where a protracted course of ECT proved life-saving in spite of prolonged duration of seizures and delayed response to treatment. The case illustrates the safety and efficacy of ECT in children. Relevant literature is also reviewed along with the case report. PMID- 12571388 TI - The role of the ombudsman in biomedical journals. PMID- 12571389 TI - A knowledge-management model for clinical practice. PMID- 12571390 TI - A case with diarrhoea, hypotension, wasting and weight loss. PMID- 12571391 TI - Spider nevus. PMID- 12571392 TI - Erythema elevatum diutinum. PMID- 12571393 TI - Coronary sinus thrombosis. PMID- 12571394 TI - Gradenigo's syndrome: findings on computed tomography and magnetic resonance imaging. PMID- 12571395 TI - Evaluating and treating anxiety disorders in medical settings. AB - Anxiety disorders and medical illness present to the primary care physician as a common comorbidity. This article aims to review the literature on the prevalence of anxiety disorders in patients presenting to primary care physicians; to address the key issues in assessing the comorbid condition; and to discuss psychological and pharmacological treatment options for patients with a comorbid anxiety disorder and medical illness. Anxiety disorders are highly prevalent within the primary care population, and these disorders significantly impact the patient's course and outcome. Fortunately, primary care physicians have a variety of effective cognitive, behavioral and pharmacological interventions available for managing these patients with comorbid anxiety and medical illnesses. PMID- 12571396 TI - Attributing death to cancer: cause-specific survival estimation. AB - Cancer survival estimation is an important part of assessing the overall strength of cancer care in a region. Generally, the death of a patient is taken as the end point in estimation of overall survival. When calculating the overall survival, the cause of death is not taken into account. With increasing demand for better survival of cancer patients it is important for clinicians and researchers to know about survival statistics due to disease of interest, i.e. net survival. It is also important to choose the best method for estimating net survival. Increase in the use of computer programmes has made it possible to carry out statistical analysis without guidance from a bio-statistician. This is of prime importance in third- world countries as there are a few trained bio-statisticians to guide clinicians and researchers. The present communication describes current methods used to estimate net survival such as cause-specific survival and relative survival. The limitation of estimation of cause-specific survival particularly in India and the usefulness of relative survival are discussed. The various sources for estimating cancer survival are also discussed. As survival-estimates are to be projected on to the population at large, it becomes important to measure the variation of the estimates, and thus confidence intervals are used. Rothman's confidence interval gives the most satisfactory result for survival estimate. PMID- 12571397 TI - Methylene blue as treatment for contrast medium-induced anaphylaxis. PMID- 12571398 TI - Topical beta blockers and atrioventricular block in the elderly. PMID- 12571399 TI - Hidden diseases detected after a fall. PMID- 12571401 TI - Intramuscular lipoma of the pectoralis major muscle. PMID- 12571400 TI - Cardiac output monitoring based on thoracic electrical bioimpedance. PMID- 12571402 TI - Epidemiology of burns among married women in India. PMID- 12571403 TI - Neurosurgery in India. PMID- 12571404 TI - Misoprostol: an old drug, new indications. PMID- 12571405 TI - A comparative study of antioxidants S-allyl cysteine sulfoxide and vitamin E on the damages induced by nicotine in rats. AB - The dietary consumption of antioxidant-rich fruits and vegetables is inversely correlated with the incidence of various diseases like cardiovascular diseases and lung cancer. We have tried to find out how far the S-allyl cysteine sulfoxide (SACS) isolated from garlic (Allium Sativum L.) can combat the nicotine-induced peroxidative damage in rats. The effects have been compared with the standard antioxidant vitamin E. Administration of SACS or vitamin E (100 mg/kg) to nicotine (0.6 mg/kg) treated rats for 21 days showed decreased concentrations of thiobarbituric acid reactive substances, hydroperoxides, and conjugated dienes in liver, lungs, and heart as compared with the values found in rats treated with nicotine alone. The activities of catalase and superoxide dismutase increased. The levels of the antioxidants like vitamins A, C, and E in the liver and glutathione in all tissues increased significantly in SACS-treated or vitamin E fed rats. However, the antioxidant status was higher when vitamin E was administered as compared with SACS administered to nicotine-treated rats. PMID- 12571406 TI - Aminoglycoside-induced reversible tubular dysfunction. AB - Nonoliguric renal insufficiency is a well-known nephrotoxic consequence of aminoglycosides, although reversible tubular damage in the absence of any change in the renal function has been occasionally found. Reported herein are 2 representative cases of a reversible tubular damage due to prolonged aminoglycoside administration: a patient with a Fanconi-like syndrome of proximal tubular dysfunction and a patient with a syndrome of hypokalemic metabolic alkalosis associated with hypomagnesemia. PMID- 12571407 TI - Downregulation of CYP2D16 by ACTH in the guinea pig adrenal cortex: time course, reversibility, and mechanism of action. AB - Studies were done to investigate the actions of ACTH on the expression of CYP2D16 in the guinea pig adrenal cortex. Guinea pigs were treated with ACTH for 1, 3, or 7 days. In addition, some animals received ACTH for 7 days and were then untreated for an additional 3 or 7 days to test for reversibility of ACTH actions. ACTH treatment caused a time-dependent decrease in the rates of adrenal microsomal bufuralol metabolism, a CYP2D-catalyzed reaction; hepatic bufuralol metabolism was unaffected by ACTH. Adrenal enzyme activity was significantly reduced by ACTH within 1 day and decreased by 80% after 7 days. Western blotting and in situ hybridization analyses revealed corresponding declines in adrenal CYP2D16 protein and mRNA concentrations. Nuclear runoff assays indicated that ACTH treatment inhibited CYP2D16 expression at the transcriptional level. Adrenal 17 alpha-hydroxylase activities were increased by ACTH treatment, but CYP17 protein concentrations were not affected. Following cessation of ACTH administration, the rates of adrenal bufuralol metabolism and CYP2D16 protein and mRNA concentrations returned to control levels within 7 days. The results demonstrate that ACTH has a relatively rapid and reversible effect to inhibit adrenal CYP2D16 transcription, thereby decreasing adrenal xenobiotic metabolism. Thus, the actions of ACTH on CYP2D16 expression are opposite to those on other adrenal p450 isozymes, indicating unique regulatory mechanisms. PMID- 12571408 TI - Cardioprotective effects of extracts from Psidium guajava L and Limonium wrightii, Okinawan medicinal plants, against ischemia-reperfusion injury in perfused rat hearts. AB - The aim of this study was to determine whether the medicinal herbs growing in Okinawa and possessing a radical-scavenging activity would exert cardioprotective effects against ischemia-reperfusion injury using isolated perfused rat hearts. Effects of the aqueous extracts from Psidium guajava L. and Limonium wrightii at concentrations having an equipotent radical-scavenging activity on myocardial injury produced by global ischemia followed by reperfusion were tested and were further compared with those of quercetin and gallic acid, major antioxidative components of P. guajava L. and L. wrightii, respectively. Both extracts significantly attenuated ischemic contracture during ischemia and improved myocardial dysfunction after reperfusion. Decreases in high-energy phosphates and increases in malondialdehyde in the reperfused hearts were significantly lessened with both plant extracts. Quercetin and gallic acid also exerted similar beneficial effects. These results indicate that P. guajava L. and L. wrightii both have cardioprotective effects against myocardial ischemia-reperfusion injury in isolated rat hearts, primarily through their radical-scavenging actions. PMID- 12571409 TI - Gonadectomy eliminates endothelium-dependent diethylstilbestrol-induced relaxant effect in rat aorta. AB - The effects of gender and castration of rats on diethylstilbestrol-induced, endothelium-dependent and endothelium-independent relaxation in rat aorta strips were studied. For this, male and female control and castrated rats were used. Diethylstilbestrol elicited a concentration-dependent (1-30 micromol/l) relaxation of isolated rat aorta. The effect was significantly higher in the presence of endothelium in aorta strips of the control group and also in female as compared with male rats. This effect is NO-dependent, since it is inhibited by N(G)-methyl-L-arginine. Castration of the rats suppressed the endothelium dependent relaxation, and it was similar to that induced in the absence of endothelium. Acetylcholine-induced relaxation was not suppressed by castration. The acetylcholine-induced relaxation was decreased in aorta strips previously relaxed by diethylstilbestrol. There are no gender differences in the diethylstilbestrol-induced, endothelium-independent component of the relaxation, nor is it modified by the hormonal environment. Therefore, diethylstilbestrol induced, endothelium-dependent relaxation in rat aorta strips is modulated by the hormonal status of the rats. PMID- 12571410 TI - Self-administration of both ethanol and nicotine in rats. AB - Self-administration of either nicotine (NIC) or ethanol (ETH) has been extensively studied. This study addressed for the first time the self administration of both substances when offered together. Male and female rats of different ages were offered NIC and ETH using the two- or three-bottle free choice method. When NIC and ETH were offered together at different concentrations to young male rats (about 45 days old), intake of NIC increased with increasing NIC concentrations, and intake of ETH increased with decreasing ETH concentrations, but these effects were independent of the presence of the second drug. These rats also consumed the same amounts of NIC or ETH regardless of whether offered individually or together. A prior choice of only NIC or ETH did not affect a subsequent intake of both drugs offered together. A choice of both drugs for 24 h for several days followed by a choice for only 2 h for several days showed the same intake of NIC but a decreased intake of ETH for the shorter period. Young female rats (about 45 days old) and older male rats (about 75 days old) consumed the same amounts of NIC but less ETH than did the young male rats. These results show that young male rats voluntarily consume NIC and ETH independently of each other and that preexposure to one drug does not affect the subsequent intake of both drugs in combination. The data also suggest that these drugs act on different reward centers which have to be 'satisfied' independently of each other. PMID- 12571411 TI - Sildenafil, a phosphodiesterase-5 inhibitor, enhances the antinociceptive effect of morphine. AB - Various evidence has demonstrated a role of the nitric oxide (NO)/cGMP signaling pathway in the processing of nociception. The exact role of phosphodiesterase-5 (PDE-5) via the NO/cGMP pathway is not fully understood in pain response. The aim of the present study was to investigate the possible peripheral interaction between a PDE-5 inhibitor (sildenafil) and morphine. Carrageenan-induced hyperalgesia in rats and the acetic-acid-induced writhing test in mice were used as animal models. Local administration of sildenafil (50-200 microg/paw, i.pl.) exhibited a dose-dependent antinociceptive effect against the paw pressure test. Sildenafil also demonstrated an antinociceptive effect (1-10 mg/kg, i.p.) against in the writhing test. Co-administration of sildenafil (100 microg/paw, i.pl. and 2 mg/kg, i.p.) significantly enhanced the antinociceptive effect of morphine (2 microg/ paw, i.pl. and 2 mg/kg, i.p respectively). The antinociception produced by the drugs alone or combined was due to a local action, as its administration in the contralateral paws was ineffective. Pretreatment with N(G)-nitro-L arginine methyl ester (an NO synthesis inhibitor), methylene blue (gunalyl cyclase inhibitor) or naloxone (opioid receptor antagonist) blocked the effect of a sildenafil-morphine combination in both tests. The results suggest that opioid receptor (NO and cGMP) mechanisms are involved in the combined antinociceptive effect. Further, sildenafil produced antinociception per se and increased the response of morphine, probably through the inhibition of cGMP degradation. PMID- 12571412 TI - Possible involvement of prostaglandins F(2alpha) and D(2) in acetylcholine induced positive inotropy in isolated mouse left atria. AB - The inotropic action of prostaglandins PGF(2alpha), PGD(2) and PGE(2) on isolated mouse left atria was characterized and compared with the positive inotropic action of acetylcholine, which has previously been shown to be mediated by prostaglandins released from the endocardial endothelium. PGF(2alpha), PGD(2) and PGE(2) produced positive inotropic responses; the time course of the change in contractile force induced by PGF(2alpha) and PGD(2) was about the same as that by acetylcholine, while that by PGE(2) was slower. Fluprostenol and sulprostone, FP and EP receptor agonists, respectively, had positive inotropic effects while BW 245C, a DP receptor agonist, had no effect. AH-6809, a DP receptor antagonist, had no inhibitory effect on the positive inotropic response to PGD(2). Dimethylamiloride, an inhibitor of Na(+)/H(+) exchange, inhibited the positive inotropic response to PGF(2alpha), PGD(2) and acetylcholine, but not PGE(2). Fluorometric pH measurement with carboxy-SNARF-1-loaded atrial myocytes revealed no change in intracellular pH on application of PGF(2alpha). PGF(2alpha) and PGD(2) significantly prolonged the duration of the atrial action potential while PGE(2) had no significant effect. These findings suggest that prostaglandins induce positive inotropic response in mouse atria through FP and EP receptor stimulation and that the former mechanism mediates in part the positive inotropic response to acetylcholine. PMID- 12571413 TI - P2X(7) receptors are redistributed on human monocytes after pore formation in response to prolonged agonist exposure. AB - The P2X(7) cell surface receptor is responsible for adenosine triphosphate dependent lysis of immune cells following the formation of non-selective membrane pores. This study examined changes in P2X(7) receptor distribution following agonist treatment. Human THP-1 monocytes were exposed to the agonist 2'-3'-O-(4 benzoyl-benzoyl) adenosine 5'-triphosphate for 1-40 min after which P2X(7) receptors were immunogold labelled. The number and distribution of P2X(7) receptors on the cell membrane were quantified using transmission electron microscopy. Increasing the time of agonist exposure resulted in clustering of P2X(7) receptors, a decrease in individual P2X(7) receptors and no change in the total number of P2X(7) receptors. These results suggest that pore formation does not involve further insertion or initial movement of receptors in the membrane but that P2X(7) receptors do cluster together after prolonged activation. PMID- 12571415 TI - Animal models of inflammatory bowel disease: an overview. AB - Inflammatory bowel disease (IBD) research has been performed in human in vitro studies and in in vivo studies using appropriate animal models. Such animal models allow both the examination of inflammatory processes (both early and late events) as well as the evaluation of new therapeutic modalities. Since the first description of the immune complex colitis in rabbits in 1961, overall 63 models have been described, most of which within the last decade. These IBD animal models can be divided into 5 different categories: (1) antigen-induced colitis and colitis induced by microbials; (2) other inducible forms of colitis (chemical, immunological, and physical); (3) genetic colitis models (transgenic and knock-out models); (4) adoptive transfer models, and (5) spontaneous colitis models. In spite of the high overall number of models, none of them is the 'perfect' model and therefore numerous aspects need to be considered when choosing one model for a particular study. Importantly, most clinical aspects (e.g. extraintestinal manifestations or fistula) have recently been described in one or the other model allowing further studies with relevance for almost all aspects of IBD. So far, IBD animal models have taught us important lessons, e.g. the requirement of T-helper cells in most models, the need of a particular genetic background, and the role of the flora in the initiation of IBD. It is expected that our understanding of IBD will further increase in a number of additional areas using animal models, e.g. exploring the role of the innate immune system in IBD. PMID- 12571416 TI - Role of commensal bacteria in chronic experimental colitis: lessons from the HLA B27 transgenic rat. AB - Rats on Lewis or Fischer background, transgenic for human HLA-B27 and beta(2) microglobulin genes spontaneously develop colitis, gastritis, arthritis, dermatitis, orchitis, epididymitis, carditis, alopecia and nail changes. Disease susceptibility correlates with the gene copy number and is influenced by the genetic background. The pathomechanism in this model is still not completely understood. Cell transfer experiments indicate an essential role of HLA-B27 expression in bone marrow-derived cells. On Fischer background the onset of colitis occurs at 2 months of age, peaks at 3 months of age, and plateaus. Histologic findings include inflammatory cell infiltration, mostly limited to the mucosa, crypt hyperplasia, reduction of goblet cells, occasionally crypt abscesses and early ulcers. There is evidence that normal luminal bacteria play an essential role in initiating and perpetuating chronic colitis and gastritis in HLA-B27 transgenic rats: Transgenic rats raised under germ-free conditions do not develop gastrointestinal disease, whereas transgenic littermates exposed to specific pathogen-free bacteria develop colitis and gastritis within 2-4 weeks. Obligate anaerobic bacteria, especially Bacteroides spp., may play a predominant role since metronidazole prevents colitis and transgenic germ-free rats contaminated with a cocktail of six obligate and facultative anaerobic bacteria develop colitis and gastritis only in the presence of Bacteroides vulgatus. Luminal bacteria may also be involved in trafficking and homing of inflammatory cells into remote organs, since varying cecal bacterial composition does not only alter local inflammation but also influences gastritis. Lymphocyte transfer experiments indicate a specific response to luminal bacteria. In summary, this animal model is suitable for investigating the influence of normal luminal bacteria on the cellular immune mechanism in chronic intestinal inflammation. PMID- 12571417 TI - The interleukin-2-deficient mouse model. AB - Interleukin-2-deficient (IL-2(-/-)) mice develop colitis with striking clinical and morphological similarities to ulcerative colitis. Since transport and barrier properties are impaired in ulcerative colitis, we studied transport and barrier functions in IL-2(-/-) mice in order to gain insight for the first time into the general pathomechanisms of disturbed transport and barrier function of the intestine during inflammation. Alternating current impedance analysis was used to determine tissue conductance in the inflamed proximal colon of IL-2(-/-) mice and to discriminate between pure epithelial and subepithelial conductance. Surprisingly, epithelial conductance was not increased but diminished in IL-2(-/ ) mice compared to controls (20.2 +/- 1.3 versus 28.8 +/- 2.8 mS/cm(2)). Concomitantly, conductance of the subepithelial tissue layers was decreased in IL 2(-/-) mice as a result of edema and infiltration with inflammatory cells. In the distal colon, electrogenic Na(+) transport (J(Na)) mediated by the epithelial Na(+) channel (ENaC) was measured 8 h after stimulation with 3.10(-9) M aldosterone in vitro as the drop in I(SC) (short circuit current) after addition of 10(-4) M amiloride. In controls, J(Na) was 6.9 +/- 0.9 micromol x h(-1) x cm( 2), whereas it was abolished in IL-2(-/-) mice. In conclusion, the inflamed colon of IL-2(-/-) mice exhibits a severe disturbance in Na(+) uptake via the ENaC in the absence of a barrier defect. Thus, reduced expression of active absorptive transport and not a barrier defect is responsible for the diarrhea in this model of intestinal inflammation. This makes this model suitable for studying the general pathomechanisms of the inflammatory downregulation of intestinal transport proteins. PMID- 12571418 TI - Neutrophil migration across the intestinal epithelial barrier--summary of in vitro data and description of a new transgenic mouse model with doxycycline inducible interleukin-8 expression in intestinal epithelial cells. AB - The intestinal epithelium serves as a protective barrier between the gut lumen and the underlying mucosa. During intestinal inflammation in inflammatory bowel disease (IBD) there is increased paracellular permeability and strong migration of the neutrophils into the mucosa as well as transepithelial neutrophil migration with the formation of crypt abscesses. From in vitro data we know that neutrophil migration is triggered by epithelial-derived interleukin-8 (IL-8). A pathogen-elicited epithelial chemoattractant activity, which is released by intestinal epithelial cells from the apical surface, seems to be responsible for the final step in transepithelial migration. To mimic the pathophysiological situation with influx of neutrophils during IBD, a double transgenic mouse model with doxycycline-inducible IL-8 expression in intestinal epithelial cells was established. This model will allow to study the role of neutrophil influx during chronic intestinal inflammation. It will also provide insights into the mechanisms of neutrophil transepithelial migration. PMID- 12571420 TI - Early events in the pathogenesis of a murine transfer colitis. AB - Inflammatory bowel disease (IBD) is a chronic, disabling disease. A dysregulated immune response seems to play a pivotal role in the pathogenesis of this disorder. Here we will review current concepts of the adoptive transfer model of IBD with particular emphasis on early events in disease development. In the adoptive transfer model, the reconstitution of immunoincompetent mice with CD4+ T cells from congenic donor animals leads to severe colitis. We will address the question as to which CD4+ T cell subsets might be involved in the induction, suppression, or regulation of disease, and review data concerning the specificity of their T cell receptor and its putative MHC restriction elements. We will also discuss whether and at what anatomical sites donor T cells could be primed in the recipient. PMID- 12571419 TI - Role of gamma delta T cells in inflammatory bowel disease. AB - gammadelta T cells have previously been shown to play a protective role in various animal models of chronic inflammation (e.g., experimental autoimmune encephalomyelitis, collagen-induced arthritis, and non-obese diabetes). This immunoregulatory potential is exerted by synthesizing various anti-inflammatory cytokines and growth factors (e.g., transforming growth factor-beta). As the normal balance between inflammatory and regulatory cytokines is perturbed in inflammatory bowel disease (IBD) a protective effect of gammadelta T cells seems likely. This notion is supported by our finding of increased mortality of rats with 2,4,6-trinitrobenzene sulfonic acid-induced colitis following gammadelta T cell depletion. In contrast, no effect was observed after depletion of gammadelta T cells in a Crohn's disease animal model with terminal ileitis (TNF(DeltaARE) mice). Therefore, future studies must further define where in the intestinal immune system gammadelta T cells exert their protective function and how this can be used in the treatment of IBD. PMID- 12571421 TI - Anti-interleukin-18 therapy in murine models of inflammatory bowel disease. AB - Interleukin (IL)-18 is a cytokine with a broad array of effector functions, the most prominent of which is to act synergistically with IL-12 in interferon-gamma production and the induction of a strong T-helper-1-mediated immune response. In addition, IL-18 also upregulates the production of proinflammatory cytokines such as IL-1 and tumor necrosis factor-alpha. Analysis of IL-18-deficient mice revealed an important role of IL-18 in the activation of macrophages and natural killer cells in the context of infection with intracellular bacteria or parasites. In humans, it was reported that IL-18 is elevated at sites of inflammation in inflammatory bowel disease (IBD), particularly in Crohn's disease, suggesting a possible role for IL-18 in the development and persistence of IBD. In this review we summarize recent findings on the functional role of IL 18 in the pathogenesis of colitis with a special focus on murine models of IBD. The neutralizing mouse anti-mouse IL-18 antibodies generated in our group should facilitate the evaluation of the efficiency of therapeutic blockade of endogenous IL-18 in chronic mouse models of colitis besides the use of recombinant forms of the inhibitory IL-18-binding protein. PMID- 12571422 TI - A new model of chronic colitis in SCID mice induced by adoptive transfer of CD62L+ CD4+ T cells: insights into the regulatory role of interleukin-6 on apoptosis. AB - OBJECTIVE: The proinflammatory cytokine interleukin (IL)-6 is involved in various chronic inflammatory processes. IL-6 is a predominant cytokine produced by lamina propria T cells in Crohn's disease and experimental colitis. This study was designed to examine the effect of a neutralizing IL-6-receptor (IL-6R) antibody on the programmed cell death of mucosal T cells in the CD62L+ CD4+ SCID transfer model of chronic experimental colitis in mice and to gain more insight into the pathogenesis of this transfer colitis model. METHODS: For adoptive transfer, we isolated CD62L+ CD4+ double-positive T cells from wild-type BALB/c mice followed by intraperitoneal application of 1 million cells in CB17 SCID mice. The purity of the transferred T-cell population was tested by FACS analysis. Cytokine secretion was measured by ELISA. Western blot analysis was performed to detect phospho-STAT-3 in protein extracts of splenic cells. Cryo- and paraffin colon cross-sections were used to perform immunochemical or fluorescence TUNEL stainings. RESULTS: We isolated CD62L+ CD4+ and CD62L- CD4+ T cells. In vitro studies showed an increased production of IL-4 by CD62L- CD4+ T cells compared to CD62L+ T cells. 8-10 weeks after transfer of CD62L+ CD4+ T cells in SCID mice, reconstituted mice developed wasting disease, anal prolapse and diarrhea, whereas mice reconstituted with CD62L- CD4+ did not, similar to anti-IL-6R-treated CD62L+ CD4+-reconstituted SCID mice. Anti-IL-6R-treated reconstituted SCID mice showed decreased levels of activated STAT-3. The previously described efficacy of anti IL-6R antibody treatment on colitis activity appeared to be due to the induction of apoptosis, as many TUNEL-positive cells were detected in the lamina propria. CONCLUSIONS: These results suggest that the activation of the IL-6/STAT-3 signaling pathway plays a pivotal role in the pathogenesis of CD62L+ CD4+ transfer colitis. Moreover, the application of a neutralizing antibody to IL-6R induces apoptosis in transferred T cells. These data implicate the importance of anti-apoptotic pathways in chronic disease and might contribute to future therapies. PMID- 12571423 TI - Interleukin-12 antagonists as new therapeutic agents in inflammatory bowel disease. AB - Inflammatory bowel diseases (IBDs; Crohn's disease (CD) and ulcerative colitis) are chronic inflammatory diseases leading to destruction of gastrointestinal tissue. They are characterized by an exaggerated immune response. In CD, an increased expression of T-helper-1 (Th1) cytokines was observed in which interleukin-12 (IL-12) seems to play a pivotal role. Different immunosuppressive agents have been used to treat patients suffering from IBD, nevertheless remarkable side effects or treatment failure are limiting factors in this regard. Therefore, studies on more specific treatment of CD have recently been published, using recombinant anti-inflammatory cytokines or inhibitors of proinflammatory cytokines and their receptors. Beyond these principles anti-IL-12 strategies seem to play a promising role because of the central position of this Th1-inducing cytokine in the inflammatory cascade. Up to now anti-IL-12 antibodies, complement receptor-3 antibodies and IL-12p40 homodimers have been evaluated in their potential to suppress the mucosal inflammation. Based on our understanding of the pathogenesis of CD, the available data and experiences concerning these principles are presented in this review. PMID- 12571424 TI - Functional involvement of CD44 variant 7 in gut immune response. AB - A major problem in inflammatory bowel disease (IBD) is the accumulation of highly activated T-helper cells that are refractory to apoptosis induction. Hence, persistent inflammatory lesions are prevalent and are the basis of chronic disease. In IBD upregulation of costimulatory molecules on lamina propria lymphocytes has been described leading to apoptosis resistance. CD44 is a cell adhesion molecule and a signalling receptor that functions as a costimulatory molecule in T-cell activation. Several variant isoforms of CD44 (CD44v) are expressed by alternative splicing of variant exons encoding extracellular regions. Particularly isoforms containing CD44v7 are expressed on T cells and macrophages in T-helper-1 (Th1)-mediated chronic inflammation and autoimmune diseases. In this review recent data on the functional involvement of CD44v7 isoforms in IBD are discussed. In a mouse model of experimental colitis blockade or deletion of CD44v7 protects mice from severe intestinal inflammation by inducing apoptosis in lamina propria mononuclear cells. Recently, we observed that in lamina propria mononuclear cells from the inflamed but not uninflamed mucosa of patients with Crohn's disease, blockade of CD44v7 isoforms also induces apoptosis. The finding that obstruction of CD44v7 isoforms can antagonize Th1 cytokine-dependent immune pathology identifies CD44v7 as a target in the treatment of inflammatory diseases such as IBD, rheumatoid arthritis, multiple sclerosis and other autoimmune diseases in which CD44v7 isoforms are upregulated. PMID- 12571425 TI - The Stanley Foundation Bipolar Network: results of the naturalistic follow-up study after 2.5 years of follow-up in the German centres. AB - The Stanley Foundation Bipolar Network (SFBN) is an international, multisite network investigating the characteristics and course of bipolar disorder. Methods (history, ratings and longitudinal follow-up) are standardized and equally applied in all 7 centres. This article describes demographics and illness characteristics of the first 152 German patients enrolled in the SFBN as well as the results of 2.5 years of follow-up. Patients in Germany were usually enrolled after hospitalisation. More than 72% of the study population suffered from bipolar I disorder and 25% from bipolar II disorder. The mean +/- SD age of the study participants was 42.08 +/- 13.5 years, and the mean +/- SD age of onset 24.44 +/- 10.9 years. More than 40% of the sample reported a rapid-cycling course in history, and even more a cycle acceleration over time. 37% attempted suicide at least once. 36% had an additional Axis I disorder, with alcohol abuse being the most common one, followed by anxiety disorders. During the follow-up period, only 27% remained stable, 56% had a recurrence, 12.8% perceived subsyndromal symptoms despite treatment and regular visits. 27% suffered from a rapid-cycling course during the follow-up period. Recurrences were significantly associated with bipolar I disorder, an additional comorbid Axis I disorder, rapid cycling in history, a higher number of mood stabilizers and the long-term use of typical antipsychotics. Rapid cycling during follow-up was only associated with a rapid cycling course in history, a higher number of mood stabilizers and at least one suicide attempt in history. PMID- 12571426 TI - Electronic diary for bipolar patients. AB - Long-term monitoring methods providing an overview of the course of bipolar disorder of individual patients are a clinical necessity at least for patients who require a combination therapy with drugs that have only proven their efficacy for monotherapy. The Life Chart Method (LCM) of the NIMH is an adequate method for this purpose. Unfortunately, due to data entry and management requirements, it is too expensive for everyday clinical use. The 'electronic diary for patients with bipolar disorder' is meant to provide a method to minimize the effort for detailed long-term monitoring of patients with bipolar affective disorder and thus make it available for the everyday clinical use for every bipolar patient. PMID- 12571427 TI - Using Health Belief Models to understand the efficacy-effectiveness gap for mood stabilizer treatments. AB - OBJECTIVE: This study explored the utility of Health Belief Models (HBM) in explaining medication adherence in subjects with severe affective disorders. METHOD: Well-established measuring instruments, with confirmed reliability and validity, were used to assess each component of two HBMs and adherence to mood stabilising medication in 98 subjects with either bipolar (805 subjects) or unipolar disorders. RESULTS: About 30% of subjects met criteria for partial adherence to medication. Demographic and diagnostic variables did not predict adherence status. Subjects' beliefs about themselves and their control over the disorder were more important than side-effects in predicting medication adherence. Self-reported assessments were predictive of admission to hospital in the year after the interview. CONCLUSION: Although the study has a number of methodological limitations, the results suggest that clinical assessment of components of HBMs may improve the detection of patients at risk of non-adherence to mood-stabilising medication. PMID- 12571428 TI - Neuropsychological performance in depressed and euthymic bipolar patients. AB - INTRODUCTION: Recent studies have suggested that the presence of persistent cognitive dysfunctions in bipolar patients is not restricted to acute episodes, but they persist even during remission states. Nevertheless, there are several methodological pitfalls in most studies, such as unclear remission criteria, diagnostic heterogeneity or small sample sizes. PATIENTS AND METHODS: Several domains of cognitive function were examined in 30 depressed bipolar patients [DSM IV criteria for major depression, Hamilton Depression Scale (HDRS) > or =17] and 30 euthymic bipolar patients (at least 6 months of remission, HDRS < or =8 and Young Mania Rating Scale, YMRS < or =6). Psychosocial functioning was assessed through General Assessment of Functioning. RESULTS: The two groups showed a similar pattern of neuropsychological performance. However, the depressed group was significantly impaired on the Controlled Oral Word Association Test, FAS (COWAT), a measure of verbal fluency, compared with the euthymic group. On the other hand, functional outcome in euthymic patients was related to verbal fluency, even after controlling for residual depressive symptoms. CONCLUSIONS: Neuropsychological performance was similar in both groups, except for verbal fluency, which was lower in the depressed group. Poor verbal fluency was related to a poor social outcome in euthymic patients. Further research including longitudinal designs aimed at evaluating changes in cognition in these patients is warranted. PMID- 12571429 TI - Initial lithium and valproate combination therapy in acute mania. AB - Early onset of treatment efficacy is especially important for severe mania syndromes. Lithium has usually a delayed onset of response, which is disadvantageous for inpatient treatment of severe mania. Valproate is effective in treating acute mania and has sedative properties. It has, however, a response rate of approximately only two thirds. Therefore, the initial combination of valproate and lithium was evaluated in a prospective case series of 12 patients, and a retrospective analysis was carried out for 5 patients in comparison with a pretreatment period when lithium therapy was applied without valproate. Outcome criterion for analysis was the latency of response and remission, as well as the amount of neuroleptics used for additional sedation. Patients had a mania syndrome-severity score comparable with the mean mania score of the bipolar manic patients treated in the same unit. All patients under the combination treatment responded. The response occurred within a shorter time compared with the lithium pretreatment episode, which is statistically marginally significant, and the use of neuroleptic medication could be markedly reduced from 18,601.6 mg chlorpromazine equivalents to 3,927.6 mg (p < 0.025). The initial valproate lithium combination therapy seems to be a safe and effective way to treat severe mania syndromes in the clinic. PMID- 12571430 TI - Severe tardive dyskinesia in affective disorders: treatment with vitamin E and C. AB - Tardive dyskinesia caused by antipsychotic treatment is a severe problem not only in the management of schizophrenia, but also of affective disorders. Vitamin E monotherapy has been used in schizophrenic patients with tardive dyskinesia. Pharmacologists warn against high dosage of vitamin E because of its pro oxidative effects on low-density lipoprotein with consecutive cardiac risks. Addition of vitamin C probably reduces this risk because of its interactions with vitamin E, i.e. vitamin C reduces vitamin E radicals formed when vitamin E scavenges the oxygen radicals. We have therefore tested the safety and efficacy of combining vitamin C and E in a sample of patients with affective disorders and tardive dyskinesia who had previously been treated with antipsychotics due to psychotic symptoms. In all 6 patients, a reduction of tardive symptomatology was seen. In our sample, no side effects were observed. Further studies on this combination therapy are suggested. PMID- 12571431 TI - Nefazodone in psychotic unipolar and bipolar depression: a retrospective chart analysis and open prospective study on its efficacy and safety versus combined treatment with amitriptyline and haloperidol. AB - Although atypical antipsychotics are on the rise, traditional treatment of psychotic (or delusional) depression mostly includes the addition of classical antipsychotics to antidepressants. As there are only few data supporting this approach compared with antidepressant monotherapy, and almost no data comparing it with antidepressants of the latest generation, we conducted a retrospective chart analysis and a prospective, randomized open study on the efficacy and tolerability of nefazodone monotherapy versus combined treatment with amitriptyline and haloperidol in psychotic depression. The results suggest that the addition of classical antipsychotics should be reserved for those with very severe psychotic symptoms, but may not be needed in milder forms. PMID- 12571432 TI - Effectiveness of hormonal tourniquet by vasopressin during myomectomy through vasopressin V1a receptor ubiquitously expressed in myometrium. AB - To elucidate the mechanism by which myometrial vasopressin injection promotes hemostasis during myomectomy, we examined the expression of vasopressin V1a receptor transcripts in the myometrium. Vasopressin V1a receptor expression was ubiquitous, and the transcripts were detected in the myometrium not only of cycling and pregnant patients, but also of postmenopausal or GnRH agonist-treated patients. Based on these observations, we applied intraoperative myometrial vasopressin injection during myomectomy in a non-randomized study in a total of 84 patients. Vasopressin injection significantly reduced the intraoperative blood loss and postoperative hemoglobin fall in patients without and with GnRH agonist pretreatment. No serious complications occurred on account of the vasopressin injection. We conclude that intraoperative vasopressin injection is effective as a hormonal tourniquet even in GnRH agonist-pretreated myomectomy. PMID- 12571433 TI - CA 125 and vascular endothelial growth factor in the differential diagnosis of epithelial ovarian tumors. AB - Tumor markers have been investigated in differentiation of benign and malignant tumors. We analyzed CA 125 and vascular endothelial growth factor (VEGF) levels in serum and cyst fluid in patients with epithelial ovarian tumors. Serum and tumor cyst fluid of 50 patients with ovarian epithelial tumors (7 malignant, 3 bordeline and 40 benign) were assayed for VEGF by ELISA and CA 125 levels by chemoluminescence. CA 125 serum levels were significantly higher in patients with malignant and borderline tumors than in patients with benign cysts (p = 0.0005). CA 125 cyst fluid contents were comparable for malignant, borderline and benign ovarian tumors (p = 0.39). Significantly higher levels of VEGF were present in cyst fluid for malignant and borderline tumors compared with benign cysts (p < 0.0001); however, serum levels of VEGF were similar among all patients (p = 0.25). The CA 125 serum levels correlated with matched VEGF cyst fluid levels (r = 0.44, p = 0.0015). Serum CA 125 and cystic VEGF were good methods to differentiate benign and malignant epithelial ovarian tumors. Patients with elevated intracystic VEGF levels presented significantly higher CA 125 serum levels, although the CA 125 intracystic content overlapped. The angiogenesis and enhancement of vascular permeability induced by VEGF represents a new hypothesis for the release of the CA 125 antigen into the circulation in patients with ovarian epithelial neoplasm. PMID- 12571434 TI - Iron status and iron deficiency anaemia in adolescents in a Tanzanian suburban area. AB - OBJECTIVES: To assess the extent and degree of anaemia in adolescents in a Tanzanian suburban area, to estimate the contribution of iron deficiency, using serum (S)-ferritin and soluble transferrin receptor (sTfR) as markers of iron deficiency. MATERIALS: Consecutive primigravidae at booking for antenatal care (n = 76), primary school boys (n = 101) and postmenarchal girls (n = 130) age >or=12 years were investigated. METHODS: Weight and height were measured; venous blood was drawn for haematological analyses, malaria screening, S-ferritin, sTfR, and C reactive protein. Stool specimens were analysed for intestinal parasites. RESULTS: Anaemia (Hb <105 g/l) was highly prevalent in adolescent primigravidae (75.5%). Adolescent girls were more anaemic (Hb <120 g/l) than boys (14.5 vs. 7.9%). Iron deficiency and hookworm infestation were predominant in both groups of adolescents, however, malaria contributed more to anaemia in the primigravidae. Nearly 40% of the anaemic primigravidae had indication of infection, and S-ferritin was less useful as a marker of iron deficiency in this group. sTfR identified iron deficiency in both pregnant and non-pregnant adolescents. PMID- 12571435 TI - Activated protein C resistance in normal and pre-eclamptic pregnancies. AB - OBJECTIVES: A lower ratio in the classic activated protein C resistance (APC-R) test has been reported during pregnancy, which has been called 'acquired' APC-R. However, little is known about the cause of the lowered ratio, and whether or not there is a correlation with blood coagulation activation. The primary objective of our study was to determine changes in APC-R levels in each of the trimesters of normal pregnancy. The secondary objective was to confirm whether APC-R levels were lower in pregnancies complicated by pre-eclampsia than in a control group. Finally, this prospective study was performed to investigate the prevalence of APC-R among pregnant women and to elucidate its obstetric consequences. METHODS: We enrolled 35 healthy pregnant women and 47 pregnant women affected by pre eclampsia in our study. The following laboratory tests were performed: prothrombin time, partial thromboplastin time, fibrinogen levels, antithrombin III, plasmatic fibronectin (as a marker of endothelial damage), haptoglobin (as a marker of intravascular haemolysis), a functional test for APC-R and analysis of factor V Leiden mutation by polymerase chain reaction. RESULTS: The activated protein C sensitivity ratio was lower in the pathological group than in the control group (p = 0.008 and p = 0.02, respectively). Plasmatic fibronectin was found to be higher in the pathological group than in the control group (p = 0.05). Finally, the overall prevalence of factor V Leiden mutation was 5.4%, i.e. 2/35 women (5.7%) in the control group and 3/47 women in the pathological group (6.38%). CONCLUSIONS: The APC ratio decreased after 20 weeks of gestation until week 42. This decrease was most pronounced in the third trimester, in which resistance was demonstrated in 34.2% of control group patients. In pre-eclampsia, we found a greater reduction of the APC ratio than in controls. We hypothesise that this is due to a decrease in the plasmatic levels of coagulation inhibitors and an increase in coagulatory factors. PMID- 12571436 TI - Studies on the human leukocyte antigen class I antigens in Japanese patients with macroscopically diagnosed endometriosis. AB - An increasing number of reports suggest that endometriosis is associated with abnormal immune function, although the etiology of the disease remains undefined. The human leukocyte antigen system (HLA) is known to play a role in the etiology of a number of diseases. This study examines the possible association between the HLA system and endometriosis. Fifty-five patients diagnosed with endometriosis by laparoscopic examination were typed for HLA class I antigens - HLA-A, -B, and -C antigens - using the standard microlymphocytotoxity technique of Terasaki. The frequency of HLA-B 54 and that of HLA-Cw7 were significantly higher in the patient population than in the control population. No significant difference was found between the two groups in the frequencies of the other HLA-class I antigens. The results of this study may implicate the HLA system in the development of endometriosis. PMID- 12571437 TI - Transvaginal bone anchors in female stress urinary incontinence: poor results. AB - OBJECTIVE: This study evaluates the results of a minimally invasive technique for correcting female stress urinary incontinence by transvaginal implantation of pubic bone anchors. PATIENTS AND METHODS: Female stress urinary incontinence was treated by fixing a gelatin-coated Dacron sling between two miniature titanium anchors with Prolene sutures. RESULTS: A total of 26 patients (median age 57.2 years) underwent the sling procedure. The follow-up examination was performed after 11.4 months on average. Stress incontinence showed a median improvement from grade 2 to grade 0.5 (p = 0.01), although only 16 of the 26 patients were completely continent. Urethral pressure and functional length were not significantly influenced. Impaired vaginal wound healing was seen in 14 of the 26 patients (53.8%), and 13 of them underwent revision. All patients affected (15/26, 57.7%) as well as 1 with uneventful healing showed sensory urge symptoms or detrusor instability (7/26, 26.9%). The correlation between impaired wound healing and detrusor instability was highly significant (p < 0.003). 17 of the 26 patients (65.3%) were dissatisfied or very dissatisfied with the intervention. The unfavorable results did not significantly correlate with the patients' age, the number of previous operations, or the surgeon's skill. CONCLUSION: In view of the poor vaginal wound healing and the resultant irritative symptoms, transvaginal bone anchoring with fixation of a Dacron sling must be regarded as an unsuitable technique. PMID- 12571438 TI - Fetal blood saturation during the 1st and 2nd stage of labor and its relation to the neonatal outcome. AB - OBJECTIVE: The aim of this study was to evaluate fetal blood oxygenation (SpO(2)) by means of continuous pulse oximetry during labor and its relation to the neonatal outcome. MATERIALS AND METHODS: Fetal SpO(2) was measured continuously during labor with a noninvasive pulse oximetry for fetal application. The average, minimum and maximum SpO(2) levels were evaluated separately for the 1st and 2nd stage of labor. The average SpO(2) of the fetus was compared to the neonatal outcome assessed by the levels of pH, pO(2) and pCO(2) in the fetus' umbilical blood and to the Apgar score. RESULTS: Twenty-eight patients were monitored by fetal pulse oximetry. All the patients had normal, vaginal delivery. During the 1st stage of labor, the average fetal SpO(2) was 51.78 +/- 8.00%, the minimum SpO(2) level was 37.61 +/- 9.86%, and the maximum level of SpO(2) was 63.82 +/- 7.37%; in the 2nd stage of labor, the average SpO(2) level was 44.91 +/ 8.28%, the minimum level was 35.00 +/- 9.22%, and the maximum SpO(2) was 52.30 +/- 9.36%. A significant decrease in the fetal average and maximum SpO(2) levels was observed between the 1st and the 2nd stages of labor (the average SpO(2) was 51.78 +/- 8.00% vs. 44.91 +/- 8.28%, p = 0.00029; the maximum SpO(2) was 63.82 +/ 7.37% vs. 52.30 +/- 9.36%, p < 0.00001). A significant correlation between the average SpO(2) level during the 1st and 2nd stage of labor and the Apgar score at the first minute of outcome was observed (R = 0.43, p = 0.031). No relationship between the fetal SpO(2) during the 1st and the 2nd stage of labor and the pH, pCO(2), and pO(2) in the fetal umbilical blood were observed. CONCLUSIONS: During the 2nd stage of labor, a significant decrease in the fetus' SpO(2) can be observed. The fetus' SpO(2) level >30% in the 1st and 2nd stage of labor was related to the good neonatal outcome. The assessment of the fetal SpO(2) during the 1st stage of labor seems to be important in predicting neonatal outcome. PMID- 12571439 TI - Serum adenosine deaminase activity in women with pre-eclampsia. AB - The present study investigated serum adenosine deaminase (ADA) activity and the patterns of two ADA isoenzymes, ADA1 and ADA2, and to evaluate the possible role of cell-mediated immunity as causes of the changes in ADA activity in pre eclampsia. We measured serum activities of total ADA, ADA1 and ADA2 in pre eclampsia (n = 22) and normal pregnancy (n = 22). Peripheral blood monocyte counts and neopterin levels, reflecting the activation of the monocyte-macrophage cell system, were also measured. In pre-eclampsia, serum total ADA and ADA2 activities were significantly increased compared with normal pregnancy (p < 0.05), which were accompanied by increases in serum neopterin levels. These results suggest that increased total ADA activity reflects increases in ADA2 activity, which may be in part related to enhanced cell-mediated immunity during pre-eclampsia. PMID- 12571440 TI - Plasma 5'-nucleotidase activities and uric acid levels in women with pre eclampsia. AB - The present study investigated plasma activity of 5'-nucleotidase, a key enzyme in the production of adenosine, in pre-eclampsia, and evaluated the relationship between changes in 5'-nucleotidase activity, and levels of uric acid, endproduct of the purine metabolism, and the severity of pre-eclampsia. We measured plasma 5'-nucleotidase activities and uric acid levels in women with 18 normal pregnancies, mild and severe pre-eclampsia. In mild and severe pre-eclampsia, plasma 5'-nucleotidase activities and uric acid levels were significantly increased compared with those in normal pregnancy (p < 0.05). Plasma 5' nucleotidase activity increased according to increases in uric acid levels and the severity of pre-eclampsia. These results suggest that increased plasma 5' nucleotidase activity may, at least in part, be related to changes in purine metabolism in pre-eclampsia. PMID- 12571441 TI - Is the sampling site along the umbilical artery significant? AB - The purpose of umbilical blood sampling is to obtain accurate reflection of fetal acid-base status at birth. The site along the umbilical artery from which blood should be sampled postpartum was not specified. We evaluated if blood gases and pH values from three sampling sites along the umbilical artery are different. After defining the range of intraobserver (method) variability, blood pO(2), pCO(2), and pH were directly determined. The data showed consistent and significant increases in arterial pH and pCO(2) values and decreases in pO(2) values from near the fetal cord insertion to the placenta. The largest difference was noted between the fetal site and the placental plate and the smallest between the site near the placental cord insertion and the placental plate. We conclude that the site of cord blood sampling should be standardized and the umbilical artery should be sampled at a site nearest to the neonate. PMID- 12571442 TI - High-dose oral misoprostol for mid-trimester pregnancy interruption. AB - OBJECTIVE: To evaluate the efficacy of high-dose oral misoprostol for mid trimester pregnancy interruption. METHODS: We reviewed our experience with high dose oral misoprostol for mid-trimester pregnancy interruption from November 1995 to May 1999. Patients undergoing labor induction for intrauterine fetal demise or medically indicated pregnancy termination at 13-32 weeks of gestation with a non dilated cervix were evaluated. Patients received 400 microg misoprostol orally every 4 h. Women undelivered within 24 h were considered failures and were treated with high-dose oxytocin as previously described. For comparison, a group of women treated with high-dose oxytocin were evaluated. RESULTS: Forty-seven pregnancies were managed with misoprostol (n = 23) or high-dose oxytocin regimen (n = 24). Both groups were similar with respect to induction indication, gestational age, maternal age/parity, laminaria use, and initial cervical dilation. Induction-to-delivery interval (mean +/- SD) was significantly shorter in the misoprostol cohort (15.2 +/- 6.7 h) compared with those treated with oxytocin (21.7 +/- 11.0 h; p = 0.02). Additionally, a significantly greater percentage of women treated with misoprostol delivered within 24 h (91.0%) compared with the oxytocin group (62.0%; p = 0.04). Adverse outcomes and side effects were not significantly different between the study groups. CONCLUSION: High-dose oral misoprostol is more effective than concentrated oxytocin infusion for mid-trimester pregnancy interruption. PMID- 12571443 TI - Primary jejunal leiomyosarcoma mimicking a gynecologic tumor. AB - Alimentary leiomyosarcoma (LMS) is not uncommon; however, LMS which metastasizes to the ovary and uterine serosa is extremely rare. We present a case diagnosed as LMS due to the negative finding on the CD117 stain, which is the most specific criterion for the differential diagnosis of gastrointestinal stromal tumor and LMS. Despite the daunting reoperation which was performed within a short interval in this case, the patient made a rapid recovery from both procedures. PMID- 12571444 TI - Severe allergy in a pregnant woman after vaginal examination with a latex glove. AB - Recently case reports about latex allergy were increased. We have experienced severe latex allergy in a pregnant women after vaginal examination with a latex glove. A 33-year-old woman, 38 weeks pregenant, was hospitalized for management of fetal IUGR. She underwent a vaginal examination with a latex glove and soon developed severe anaphylactic reactions. Although two hours later her condition had remarkably improved, regular uterine contractions appeared, and fetal cardiotocogram showed late decelerations. So emergency cesarean section was performed under the situation without using any latex products. She delivered a male infant weighing 2,227 g (-2.21 SD) at 38 weeks gestation. His Apgar score was 5 points at 1 min and 8 points at 5 min. The latex-specific IgE of this patient was high and IgE for banana, avocado and kiwi were also positive. These foods showed cross-reactions with latex, but she had no history of allergic reactions against these foods. She had a history of atopic reactions and of atopic dermatitis while working as a nurse at the ICU. It is thought that this patient was in the high-risk group. PMID- 12571445 TI - Astrocytes and brain injury. AB - Astrocytes are the most numerous cell type in the central nervous system. They provide structural, trophic, and metabolic support to neurons and modulate synaptic activity. Accordingly, impairment in these astrocyte functions during brain ischemia and other insults can critically influence neuron survival. Astrocyte functions that are known to influence neuronal survival include glutamate uptake, glutamate release, free radical scavenging, water transport, and the production of cytokines and nitric oxide. Long-term recovery after brain injury, through neurite outgrowth, synaptic plasticity, or neuron regeneration, is influenced by astrocyte surface molecule expression and trophic factor release. In addition, the death or survival of astrocytes themselves may affect the ultimate clinical outcome and rehabilitation through effects on neurogenesis and synaptic reorganization. PMID- 12571446 TI - Stroke damage in mice after knocking the neutrophin-4 gene into the brain-derived neurotrophic factor locus. AB - Neurotrophins play a protective role during cerebral ischemia, and mice lacking both alleles for neurotrophin 4 (Nt4-/- ) or deficient in a single allele for brain-derived neurotrophic factor (Bdnf+/-) have increased susceptibility to cerebral ischemia. This study directly compared the biologic activities of brain derived neurotrophic factor (BDNF) and NT4 by replacing the coding sequence with the Nt4 sequence (Bdnf +/nt4-ki ). Mice expressing one allele in place of develop 61% bigger lesions after 1-hour middle cerebral artery occlusion compared with wild-type littermates. Physiologic parameters did not contribute to ischemia susceptibility. In conclusion, NT4 is less potent than BDNF in promoting brain survival after stroke. PMID- 12571447 TI - Anti-Nogo-A antibody infusion 24 hours after experimental stroke improved behavioral outcome and corticospinal plasticity in normotensive and spontaneously hypertensive rats. AB - Nogo-A is a myelin-associated neurite outgrowth inhibitory protein limiting recovery and plasticity after central nervous system injury. In this study, a purified monoclonal anti-Nogo-A antibody (7B12) was evaluated in two rat stroke models with a time-to-treatment of 24 hours after injury. After photothrombotic cortical injury (PCI) and intraventricular infusion of a control mouse immunoglobulin G for 2 weeks, long-term contralateral forepaw function was reduced to about 55% of prelesion performance until the latest time point investigated (9 weeks). Forepaw function was significantly better in the 7B12 treated group 6 to 9 weeks after PCI, and reached about 70% of prelesion levels. Cortical infarcts were also produced in spontaneously hypertensive rats (SHR) by permanent middle cerebral artery occlusion (MCAO). In the control group, forepaw function remained between 40% and 50% of prelesion levels 4 to 12 weeks after MCAO. In contrast, 7B12-treated groups showed significant improvement between 4 and 7 weeks after MCAO from around 40% of prelesion levels at week 4 to about 60% to 70% at 7 to 12 weeks after MCAO. Treatment in both models was efficacious without influencing infarct volume or brain atrophy. Neuroanatomically in the spinal cord, a significant increase of midline crossing corticospinal fibers originating in the unlesioned sensorimotor cortex was found in 7B12-treated groups, reaching 2.3 +/- 1.5% after PCI (control group: 1.1 +/- 0.5%) and 4.5 +/- 2.2% after MCAO in SHR rats (control group: 1.8 +/- 0.8%). Behavioral outcome and the presence of midline crossing fibers in the cervical spinal cord correlated significantly, suggesting a possible contribution of the crossing fibers for forepaw function after PCI and MCAO. The results suggest that specific anti-Nogo A antibodies bear potential as a new rehabilitative treatment approach for ischemic stroke with a prolonged time-to-treatment window. PMID- 12571448 TI - Temporal profile of angiogenesis and expression of related genes in the brain after ischemia. AB - Angiogenesis is an intricately regulated phenomenon. Its mechanisms in the ischemic brain have not been clearly elucidated. The authors investigated expression of angiogenesis-related genes using a complementary DNA (cDNA) array method as well as Western blotting and immunohistochemistry, and compared these studies with a temporal profile of angiogenesis in mouse brains after ischemia. The number of vessels significantly increased 3 days after injury, and proliferating endothelial cells increased as early as 1 day. This means that angiogenesis occurs immediately after the injury. Ninety-six genes implicated in angiogenesis were investigated with a cDNA array study. It was found that 42, 29, and 13 genes were increased at 1 hour, 1 day, and 21 days, respectively. Most of the well-known angiogenic factors increased as early as 1 hour. Vessel stabilizing factors such as thrombospondins also increased. At 1 day, however, thrombospondins decreased to lower levels than in the control, indicating a shift from vascular protection to angiogenesis. At 21 days, many genes were decreased, but some involved in tissue repair were newly increased. Western blotting and immunohistochemistry showed findings compatible with the cDNA array study. Many molecules act in an orchestrated fashion in the brain after ischemia and should be taken into account for therapeutic angiogenesis for stroke. PMID- 12571449 TI - Estrogen deprivation and replacement modulate cerebral capillary density with vascular expression of angiogenic molecules in middle-aged female rats. AB - The effect of postmenopausal estrogen replacement therapy (ERT) on the risk or severity of cerebrovascular disorders is as yet unclear, and the evidence for flow preservation being a mechanism of estrogen neuroprotection remains elusive. The authors examined whether estrogen-mediated flow-preserving neuroprotective mechanisms, if any, may involve its angiogenic action. This study was conducted using middle-aged (44 weeks) female rats because of the importance of aging in cerebrovascular disease in women. Middle-aged female rats were subjected to sham operation, ovariectomy, or ovariectomy with ERT. The anatomic cerebral capillary morphology showed a significant reduction in the total capillary density in the frontal cortex after ovariectomy. This was associated with marked decreases in protein and gene expression of vascular endothelial growth factor and its angiogenic receptors in cerebral vessels, as demonstrated by immunohistochemistry and hybridization. The expression levels of both estrogen receptor (ER) subtypes, ERalpha and ERbeta, in cerebral vessels were significantly reduced after ovariectomy, but ERbeta was more dramatically downregulated as assessed by the ERbeta/ERalpha ratio. These ovariectomy-induced changes were completely prevented by ERT. Vascular endothelial growth factor appears to be a critical regulatory molecule for physiologic cerebral angiogenesis in middle-aged female rats and may play an important role in the flow-preserving neuroprotective action of estrogen through its angiogenic and antiapoptotic properties. PMID- 12571450 TI - Ionotropic glutamate receptors in cerebral microvascular endothelium are functionally linked to heme oxygenase. AB - Vasodilator effects of glutamate in the cerebral circulation are, in part, mediated by carbon monoxide (CO), which is formed from heme via the heme oxygenase (HO) pathway. The hypothesis addressed was that glutamate receptors (GluRs) in cerebral microvascular endothelium are functionally linked to HO. Using a radioligand binding and immunoblotting, GluRs were characterized in cerebral microvascular endothelial cells (CMVEC) from newborn pigs. High-affinity (80 nmol/L) reversible binding of [3H]glutamate ([ 3H]Glu) was detected in CMVEC membranes. The -methyl-d-aspartate (NMDA) receptor ligands-NMDA, quinolinic acid, (+/-)1-aminocyclopentane- -1,3-dicarboxylic acid ( ACPD), AP5, 4C3HPG, and CPP and the (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate receptor ligands-AMPA, kainic acid, quisqualic acid, DNQX, and CNQX-displaced 20% to 30% of bound [3H]Glu in CMVEC membranes. Metabotropic GluRs antagonists (4CPG, PHCC, and CPPG) did not displace bound [3H]Glu. l-Aspartate, an agonist of GluRs and glutamate transporters, displaced 80% or more of bound [3H]Glu. Ionotropic (NR1 and GluR1) and metabotropic (mGluR1alpha) GluRs were detected in CMVEC by immunoblotting. Glutamate, aspartate, ACPD, AMPA, (RS)-2 amino-(3-hydroxy-5- -butylisoxazol-4-yl)propanoic acid (ATPA), and kainate (10( 5) mol/L) increased HO-directed CO formation by isolated cerebral microvessels and by cultured CMVEC. These data in newborn pigs suggest that CMVEC express ionotropic GluRs that are functionally linked to HO. GluR-mediated increases in CO formation by vascular endothelium may result in increase in cerebral blood flow. PMID- 12571451 TI - Direct comparison of local cerebral blood flow rates measured by MRI arterial spin-tagging and quantitative autoradiography in a rat model of experimental cerebral ischemia. AB - The present study determined cerebral blood flow (CBF) in the rat using two different magnetic resonance imaging (MRI) arterial spin-tagging (AST) methods and 14C-iodoantipyrine (IAP)-quantitative autoradiography (QAR), a standard but terminal technique used for imaging and quantitating CBF, and compared the resulting data sets to assess the precision and accuracy of the different techniques. Two hours after cerebral ischemia was produced in eight rats via permanent occlusion of one middle cerebral artery (MCA) with an intraluminal suture, MRI-CBF was measured over a 2.0-mm coronal slice using single-coil AST, and tissue magnetization was assessed by either a spin-echo (SE) or a variable tip-angle gradient-echo (VTA-GE) readout. Subsequently ( approximately 2.5 hours after MCA occlusion), CBF was assayed by QAR with the blood flow indicator 14C IAP, which produced coronal images of local flow rates every 0.4 mm along the rostral-caudal axis. The IAP-QAR images that spanned the 2-mm MRI slice were selected, and regional flow rates (i.e., local CBF [lCBF]) were measured and averaged across this set of images by both the traditional approach, which involved reader interaction and avoidance of sectioning artifacts, and a whole film-scanning technique, which approximated total radioactivity in the entire MRI slice with minimal user bias. After alignment and coregistration, the concordance of the CBF rates generated by the two QAR approaches and the two AST methods was examined for nine regions of interest in each hemisphere. The QAR-lCBF rates were higher with the traditional method of assaying tissue radioactivity than with the MRI-analog approach; although the two sets of rates were highly correlated, the scatter was broad. The flow rates obtained with the whole film-scanning technique were chosen for subsequent comparisons to MRI-CBF results because of the similarity in tissue "sampling" among these three methods. As predicted by previous modeling, "true" flow rates, assumed to be given by QAR-lCBF, tended to be slightly lower than those measured by SE and were appreciably lower than those assessed by VTA-GE. When both the ischemic and contralateral hemispheres were considered together, SE-CBF and VTA-GE-CBF were both highly correlated with QAR lCBF ( P< 0.001). If evaluated by flow range, however, SE-CBF estimates were more accurate in high-flow (contralateral) areas (CBF > 80 mL. 100 g(-1). min(-1) ), whereas VTA-GE-CBF values were more accurate in low-flow (ipsilateral) areas (CBF < or= 60 mL. 100 g(-1). min(-1) ). Accordingly, the concurrent usage of both AST MRI methods or the VTA-GE technique alone would be preferred for human studies of stroke. PMID- 12571452 TI - Segmental vascular resistance after mild controlled cortical impact injury in the rat. AB - In an effort to localize the site at which increased resistance occurs after brain trauma, pial arteriole diameter and pressure were assessed after mild controlled cortical impact (CCI) injury in rats using an open cranial window technique. The authors tested the hypothesis that an increase in resistance accompanied by vasoconstriction occurs at the level of the pial arterioles within the injured cortex of the brain. At 1 hour after mild CCI injury, ipsilateral cerebral blood flow was significantly reduced by 42% compared with sham injury (n = 4; < 0.05). Pial arteriole diameter and pressure remained unchanged. Resistance in the larger arteries (proximal resistance), however, was significantly greater after CCI injury (1.87 +/- 0.26 mm Hg/[mL. 100 g. min]) compared with sham injury (0.91 +/- 0.21 mm Hg/[mL. 100 g. min]; < 0.0001). Resistance in small vessels, arterioles, and venules (distal resistance) was also significantly greater after CCI injury (1.13 +/- 0.05 mm Hg/[mL. 100 g. min]) compared with sham injury (0.74 +/- 0.13 mm Hg/[mL. 100 g. min]; = 0.0001). The authors conclude that, at 1 hour after mild CCI injury, changes in vascular resistance comprise a 53% increase in the resistance distal to the area of injury and, surprisingly, a 105% increase in resistance in the arteries proximal to the injury site. PMID- 12571454 TI - Neuroprotection by complement (C1) inhibitor in mouse transient brain ischemia. AB - The authors investigated the effect of the C1 inhibitor (C1-INH), the only known inhibitor of complement C1, in a murine model of transient focal ischemia. Ischemia was induced by intraluminal occlusion of the middle cerebral artery. After 2 hours, reperfusion was produced by removing the nylon monofilament occluding the artery. The effect of 15 U C1-INH (intravenously) was evaluated in terms of general and focal neurologic deficits, ischemic volume, neutral red staining (to identify the brain areas subject to ischemic damage), and glial fibrillary acidic protein immunoreactivity (to show astrocytic response). Forty eight hours after ischemia, C1-INH significantly improved general and focal deficits by 36% and 54%, respectively, and significantly reduced infarct volume (CI-INH, 6.69% +/- 2.93%; saline, 24.24% +/- 8.24%) of total brain. Neutral red staining further showed the strong protective effect of C1-INH in cortex, hippocampus, and striatum. Astrocyte activation induced by ischemia was not affected by C1-INH. These findings show that C1-INH displayed a potent neuroprotective action by effectively reducing ischemia-reperfusion injury. PMID- 12571453 TI - Structural and functional damage sustained by mitochondria after traumatic brain injury in the rat: evidence for differentially sensitive populations in the cortex and hippocampus. AB - The cellular and molecular pathways initiated by traumatic brain injury (TBI) may compromise the function and structural integrity of mitochondria, thereby contributing to cerebral metabolic dysfunction and cell death. The extent to which TBI affects regional mitochondrial populations with respect to structure, function, and swelling was assessed 3 hours and 24 hours after lateral fluid percussion brain injury in the rat. Significantly less mitochondrial protein was isolated from the injured compared with uninjured parietotemporal cortex, whereas comparable yields were obtained from the hippocampus. After injury, cortical and hippocampal tissue ATP concentrations declined significantly to 60% and 40% of control, respectively, in the absence of respiratory deficits in isolated mitochondria. Mitochondria with ultrastructural morphologic damage comprised a significantly greater percent of the population isolated from injured than uninjured brain. As determined by photon correlation spectroscopy, the mean mitochondrial radius decreased significantly in injured cortical populations (361 +/- 40 nm at 24 hours) and increased significantly in injured hippocampal populations (442 +/- 36 at 3 hours) compared with uninjured populations (Ctx: 418 +/- 44; Hipp: 393 +/- 24). Calcium-induced deenergized swelling rates of isolated mitochondrial populations were significantly slower in injured compared with uninjured samples, suggesting that injury alters the kinetics of mitochondrial permeability transition (MPT) pore activation. Cyclosporin A (CsA)-insensitive swelling was reduced in the cortex, and CsA-sensitive and CsA-insensitive swelling both were reduced in the hippocampus, demonstrating that regulated MPT pores remain in mitochondria isolated from injured brain. A proposed mitochondrial population model synthesizes these data and suggests that cortical mitochondria may be depleted after TBI, with a physically smaller, MPT-regulated population remaining. Hippocampal mitochondria may sustain damage associated with ballooned membranes and reduced MPT pore calcium sensitivity. The heterogeneous mitochondrial response to TBI may underlie posttraumatic metabolic dysfunction and contribute to the pathophysiology of TBI. PMID- 12571455 TI - Early magnetic resonance imaging prediction of arterial recanalization and late infarct volume in acute carotid artery stroke. AB - In patients with acute ischemic stroke, early recanalization may save tissue at risk for ischemic infarction, thus resulting in smaller infarcts and better clinical outcome. The hypothesis that clinical and diffusion- and perfusion weighted imaging (DWI, PWI) parameters may have a predictive value for early recanalization and final infarct size was assessed. Twenty-nine patients were prospectively enrolled and underwent sequential magnetic resonance imaging (1) within 6 hours from hemispheric stroke onset, before thrombolytic therapy; (2) at day 1; and (3) at day 60. Late infarct volume was assessed by T2 -weighted imaging. At each time, clinical status was assessed by the National Institutes of Health Stroke Scale (NIHSS). Twenty-eight patients had arterial occlusion at day 0 magnetic resonance angiography (MRA). They were classified into two groups according to day 1 MRA: recanalization (n = 18) versus persistent occlusion (n = 10). Any significant differences between these groups were assessed regarding (1) PWI and DWI abnormality volumes, (2) relative and absolute time-to-peak (TTP) and apparent diffusion coefficient within the lesion on DWI; and (3) day 60 lesion volume on T2 -weighted imaging. Univariate and multivariate logistic regression analysis showed that the most powerful predictive factors for recanalization were lower baseline NIHSS score and lower baseline absolute TTP within the lesion on DWI. The best predictors of late infarct size were day 0 lesion volume on DWI and day 1 recanalization. Early PWI and DWI studies and day 1 MRA provide relevant predictive information on stroke outcome. PMID- 12571456 TI - Brain uptake of the acid metabolites of F-18-labeled WAY 100635 analogs. AB - The 5-HT1A ligands [ 18F]FPWAY and [ 18F]FCWAY are metabolized to [ 18F]fluorobenzoic acid (FB) and [ 18F]fluorocyclohexanecarboxylic acid (FC), respectively. To quantify the penetration of these acids into the brain, dynamic positron emission tomography studies were performed in rhesus monkeys with [ 18F]FB and [ 18F]FC. High-performance liquid chromatography analysis of arterial blood samples showed no metabolites for [ 18F]FB, whereas [ 18F]FC was rapidly metabolized to [ 18F]fluoride. A model with one tissue compartment and vascular radioactivity was used to analyze gray matter time-activity curves. For [ 18F]FC, an additional term was added to account for [ 18F]fluoride skull spillover into the brain; this term accounted for 70% to 90% of the measured radioactivity concentration at 90 minutes. For [ 18F]FB, mean gray matter parameters were as follows: K1, 10 +/- 3 micro L. min(-1). mL(-1); distribution volume, 0.052 +/- 0.006 (mL/mL). For [ 18F]FC, the values were as follows: K1, 15 +/- 4 micro L. min(-1). mL(-1); V, 0.29 +/- 0.06 mL/mL. The values were consistent with a physiologic model that included brain-to-blood pH difference and the plasma free fraction of the acid. Simulations based on [ 18F]FCWAY human data showed that [ 18F]FC uptake produces significant biases in estimates in regions with low specific binding. These results can be used to correct the tissue [ 18F]FCWAY time-activity data for brain uptake of [ 18F]FC using the measured [ 18F]FC input function. PMID- 12571457 TI - Comments from the editor-in-chief. PMID- 12571458 TI - Minimal residual disease detection in acute lymphoblastic leukemia: real improvement with the real-time quantitative PCR method? PMID- 12571459 TI - Real-time quantitative PCR: standardized detection of minimal residual disease in pediatric acute lymphoblastic leukemia. Polymerase chain reaction. AB - PURPOSE: To develop a standardized real-time polymerase chain reaction (PCR) method of quantifying minimal residual disease (MRD) in patients with pre-B acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: In a series of 24 follow-up bone marrow (BM) samples in 11 patients (14 clonal markers), we performed real time PCR assays using one consensus and one clone-specific primer for each marker. The markers analyzed included immunoglobulin heavy chain (IgH), T-cell receptor (TCR) and TEL-AML1 rearrangements. RESULTS: We achieved a detection limit of 3.3 x 10(-5) +/- 1.2 x 10(-5) and an accurate quantitation (r = -0.99) limit of 2.0 x 10(-4) +/- 8.8 x 10(-5) blasts. Both inter- and intra-assay reproducibility were exceptional. Additionally, we found comparable results to those of a "gold standard" limiting-dilution PCR assay (r = 0.62). CONCLUSIONS: The IgH, TCR and TEL-AML1 markers can be used as targets by real-time PCR under the same cycling profile, allowing quantitation of MRD in more 95% of patients with pre-B ALL. This standardized, real-time PCR technique should simplify monitoring MRD in clinical trials. PMID- 12571460 TI - Assessment of minimal residual disease in childhood non-hodgkin lymphoma by polymerase chain reaction using patient-specific primers. AB - A feasibility study was undertaken to identify patient-specific primers (PSPs) from childhood non-Hodgkin lymphoma (NHL) specimens to detect minimal residual disease (MRD). Eleven tumor specimens were amplified using immunoglobulin heavy chain and T-cell receptor primers to identify PSPs, which were then used to evaluate staging/follow-up specimens. Disease was detected in 19 of 21 staging and 16 of 17 follow-up specimens. Among seven patients in remission by 1 month, PSPs identified MRD in follow-up specimens. This study demonstrated the feasibility of PSPs to identify disease in staging and follow-up specimens, which could be used to develop strategies for MRD analysis in a larger setting. PMID- 12571461 TI - Systemic methotrexate exposure is greater after intrathecal than after oral administration. AB - PURPOSE: To compare systemic exposure after intrathecal and oral methotrexate administration. PATIENTS AND METHODS: We analyzed red cell methotrexate polyglutamate concentrations with a sensitive radioligand-binding assay in 80 patients enrolled in the Children's Cancer Group (CCG) trial 1922 for acute lymphoblastic leukemia. Methotrexate concentrations were measured 7 days after the last doses of intrathecal and oral routes, using patients as their own controls. Intrathecal methotrexate was given on an age-adjusted schedule. Data was normalized to the actual dose received per body surface area. RESULTS: The mean red cell methotrexate concentration 7 days after the last of four weekly intrathecal doses of methotrexate was 178 pmol/mL red blood cells, which was significantly greater than the result 7 days after subsequent weekly oral methotrexate of 122 pmol/mL (P = 0.00001). Intrathecal dosing resulted in an average systemic exposure ratio of 1.7 to 1 compared with oral administration. CONCLUSION: Intrathecal methotrexate administration results in significantly greater systemic exposure than oral administration. Our data support the hypothesis that the systemic effect of intrathecal methotrexate affects ALL therapy. PMID- 12571462 TI - Multimodal therapy for the treatment of nonmetastatic Ewing sarcoma of pelvis. AB - The purpose of this study was to evaluate the outcome of patients with Ewing sarcoma (ES) of the pelvis, attempting to identify prognostic factors to select patients for more aggressive treatment. Seventy-seven patients with nonmetastatic ES of the pelvis were treated at Rizzoli Institute between 1979 and 1996. Four different protocols of chemotherapy were used successively. Two protocols consisted of VACAc, and two of VACAc plus ifosfamide and etoposide. Local treatment consisted of surgery in 5 patients, radiotherapy in 60, and surgery followed by radiotherapy in 12. Mean follow-up was 11 years (range 5-25 years). Thirty-three patients remained continuously free of disease; 43 relapsed (24 due to metastases and 19 to local recurrence and metastases); 1 died of treatment related complications. The 5- and 10-year event-free survival rates were 45% and 44%, respectively, and the 5- and 10-year overall survival rates were 48 and 44. These results are significantly worse than the ones achieved in 329 contemporary patients with extrapelvic lesions treated with the same protocols of chemotherapy (5- and 10-year event-free survival = 46% vs. 64% and 44% vs. 69%). Thus, despite associated chemotherapy, the outcome of ES localized in the pelvis remains poor, and new innovative methods for the treatment of this tumor are needed. PMID- 12571463 TI - Religiosity is associated with the use of complementary medical therapies by pediatric oncology patients. AB - PURPOSE: To determine the prevalence of complementary and alternative medicine (CAM) use in pediatric oncology patients, the types of CAM used, and the factors associated with the use of CAM. PATIENTS AND METHODS: A questionnaire regarding CAM use was administered to patients/families seen in the pediatric oncology clinic at Wake Forest University Baptist Medical Center over a 12-month period. RESULTS: Based on 195 completed questionnaires, 91 (47%) patients reported use of CAM since diagnosis. Among CAM users, the most commonly used CAM therapies were faith healing, megavitamins/minerals, massage, other dietary supplements, relaxation techniques, and herbal medicines/teas. Forty-one percent of CAM users had not discussed CAM use with their physician(s). In bivariate analysis, CAM use was not associated with age at the time of survey, time since diagnosis, sex, race, parental education, or family income. A trend was noted between CAM use and older age at diagnosis. Families who reported themselves to be "very" religious were more likely to use CAM than those that are "somewhat" or "not at all" religious. CONCLUSIONS: Use of CAM is common among pediatric oncology patients and often is not discussed with the treating physician(s). Patients from very religious families are more likely to use CAM. PMID- 12571464 TI - Self-reported initial management of childhood idiopathic thrombocytopenic purpura: results of a survey of members of the American Society of Pediatric Hematology/Oncology, 2001. AB - The purpose of this study was to update physicians' self-reported initial management practices for childhood idiopathic thrombocytopenic purpura (ITP) from an initial survey in 1997. A questionnaire was sent by e-mail in October 2001 to 753 members of the American Society of Pediatric Hematology/Oncology (ASPH/O). The questionnaire had 14 questions, based on the clinical presentation of a 5 year-old boy with ITP, a platelet count of 7,000/microL, scattered petechiae, and no mucous membrane bleeding. Two hundred eighteen (29%) surveys were returned. In response to questions regarding initial treatment, 33% of physicians said they would always administer drug therapy, 38% usually, 15% sometimes, and 14% rarely/never. When asked which agent would be used if drug treatment were prescribed, 45% reported anti-D, 33% IVIG, 20% prednisone, and 2% other regimens. Only 34% of physicians would always or usually hospitalize such a patient. Hospitalization was more likely if a physician responded that he or she would always or usually use drug therapy. Physicians who saw more ITP patients were more likely to self-report sometimes or rarely/never prescribing drug therapy. Self-reported initial management of ITP by ASPH/O members in 2001 is similar to 1997 regarding the percentage of pediatric hematologists who would not use drug therapy. Among physicians who would use drug treatment, there was an increased use of anti-D and decreased use of IVIG and prednisone. This information provides the basis for designing a randomized clinical trial to compare the effect of different management strategies on the outcomes of bleeding symptoms, side effects of therapy, costs, and quality of life. PMID- 12571465 TI - Influenza immunization practices among pediatric oncologists. AB - PURPOSE: To describe the opinions of pediatric oncologists regarding the use of influenza vaccine in children with cancer and to identify factors that influence practitioners' recommendations about influenza vaccine. MATERIALS AND METHODS: A survey was sent to members of the Children's Oncology Group to inquire about their clinical experience and practice setting, opinions regarding the use of the influenza vaccine in children with cancer, and factors that influence their recommendations. RESULTS: Of 803 pediatric oncologists identified, 434 (54%) responded. Depending on the type of tumor, 65% to 69% of pediatric oncologists indicated that they routinely recommend influenza vaccine for children being treated for cancer. Respondents were much more likely to recommend influenza vaccine for children with various types of cancer if they indicated that: 1) their practice has guidelines regarding the use of influenza vaccine (odds ratios ranging from 7.2 to 11.7); 2) influenza infection is very significant (odds ratios ranging from 1.4 to 3.7); and 3) influenza vaccine is effective (odds ratios ranging from 7.2 to 14.9). CONCLUSIONS: The majority of pediatric oncologists routinely recommend influenza vaccine for children being treated for cancer; however, a significant number of pediatric oncologists do not. Clarification of the benefit of influenza vaccine for children with cancer and the institution of practice guidelines may increase the use of the influenza vaccine among pediatric oncologists. PMID- 12571466 TI - Randomized study of IVIg and high-dose dexamethasone therapy for children with chronic idiopathic thrombocytopenic purpura. AB - PURPOSE: To investigate whether pulsed high-dose dexamethasone is more efficacious than intravenous immunoglobulin (IVIg) as treatment of symptomatic chronic idiopathic thrombocytopenic purpura (ITP) in childhood. METHODS: In a 2:1 randomized study, 23 children with chronic ITP received dexamethasone (0.6 mg/kg per day for 4 consecutive days once monthly for 6 months, n = 15) or IVIg (800 mg/kg intravenously once monthly for 6 months, n = 8). After four courses of treatment a crossover was offered to nonresponders. A total of 20 children received dexamethasone and 11 received IVIg. RESULTS: One of the 8 IVIg patients and 2 of the 15 dexamethasone patients achieved complete response, defined as a platelet count of at least 150 x 10(9)/L for more than 3 months without treatment. Two of the 15 dexamethasone patients achieved partial response, defined as a platelet count of at least 30 x 10(9)/L for more than 3 months without treatment. One of the 8 IVIg patients and 5 of the 15 dexamethasone patients discontinued treatment. Five patients crossed over from IVIg to dexamethasone (one complete response) and three from dexamethasone to IVIg (none responded). In summary, 5 of the 20 dexamethasone patients achieved a complete or partial response and 1 of the 11 IVIg patients achieved a complete response. Platelet counts of at least 30 x 10(9)/L by day 3 were reached in 9 of the 12 (75%) dexamethasone patients and all 8 (100%) IVIg children using available data. Five years after study completion, two of the three children who achieved a complete response and one of the two with a partial response to dexamethasone were in remission, as was the child with a complete response to IVIg. CONCLUSIONS: Treatment with pulsed high-dose dexamethasone is not always effective in children with chronic ITP, but it is worth trying in severe symptomatic chronic childhood ITP. PMID- 12571467 TI - The evolution of juvenile myelomonocytic leukemia in a female patient with paternally inherited neurofibromatosis type 1. AB - The most common myeloid malignancy seen in children with neurofibromatosis type 1 (NF-1) is juvenile myelomonocytic leukemia (JMML), a myeloproliferative disease. The vast majority of these children have inherited the neurocutaneous disease from an affected mother; boys are more often affected than girls. We present the rare finding of a 7-year-old girl with NF-1 who developed JMML. She inherited her NF-1 from the father. At the time of her initial presentation, clonogenic assays of bone marrow mononuclear cells did not show the spontaneous growth of granulocyte-macrophage colony-forming units or hypersensitivity to granulocyte macrophage colony-stimulating factor that is characteristic of this disorder. After 1 month, repeat evaluations of the patient's clinical and laboratory test results became fully consistent with those for a diagnosis of JMML. This illustrates the stepwise evolution of this myeloproliferative disorder in NF-1 and the importance of close follow-up and reassessment of these patients. Our case is only the second report of JMML in a girl who inherited NF-1 from her father. PMID- 12571468 TI - Prenatal presentation supports the in utero development of congenital leukemia: a case report. AB - Congenital leukemia is a rare disease developing within the first 4 to 6 weeks of life. We report a female infant born with facial mass and multiple subcutaneous nodules. The facial mass was discovered by ultrasound during a routine prenatal examination at the 36th week of gestation. Biopsies were consistent with the diagnosis of acute monoblastic leukemia (AML, FAB M5b). Cytogenetic studies showed 46 XX, t(11;19)(q23;p13.1), which is only found in acute monoblastic leukemia and involves the gene. The infant died at 12 days of age and autopsy revealed a large leukemic tumor burden in several body organs. The discovery of the facial mass prenatally and massive extramedullary leukemic burden support the notion of the in utero development of congenital leukemia. PMID- 12571469 TI - Successful treatment of inflammatory myofibroblastic tumor with malignant transformation by surgical resection and chemotherapy. AB - Inflammatory myofibroblastic tumor (IMT) is a tumor composed of myofibroblasts and a mixed inflammatory infiltrate that rarely undergoes malignant transformation. The authors present the case of a 7-year-old boy with an abdominal mass diagnosed as IMT with malignant transformation. The tumor recurred twice after attempts at resection and was initially treated with vincristine and etoposide. After a third recurrence and incomplete resection, he was treated with cisplatin, Adriamycin, and methotrexate. He is disease-free after 2 years, representing successful combined surgery and chemotherapy in the treatment of malignant IMT. The use of chemotherapy for aggressive myofibroblastic tumors is reviewed. PMID- 12571470 TI - Treatment of neuroblastoma in patients with neurocristopathy syndromes. AB - Neuroblastoma, a neoplasm of neural crest cell origin, occasionally presents in association with other neural crest abnormalities such as Hirschsprung disease, congenital central hypoventilation, autonomic disturbances, and other tumors. These associations have been termed "neurocristopathy syndromes." In the past, chemotherapy has not been considered for patients with neurocristopathy associated neuroblastoma because of their complicated medical problems. The authors describe two patients with neurocristopathy syndromes from our institution who underwent surgery and standard chemotherapy treatment of their intermediate-risk neuroblastoma. They represent two of only three long-term disease-free survivors of neurocristopathy-associated neuroblastoma reported in the literature. PMID- 12571471 TI - A pediatric case of lymphomatoid granulomatosis with onset after completion of chemotherapy for acute myeloid leukemia. AB - In this case report, we present a pediatric case of lymphomatoid granulomatosis (LG) with onset just after the completion of chemotherapy for childhood acute myeloid leukemia (AML). After the completion of maintenance therapy, the patient was admitted to our clinic with a complaint of cough. Radiologic examinations revealed nodular lesions in lungs, liver, and kidney. His bone marrow was in remission. The histopathologic examination of the open lung biopsy was consistent with LG. He received only one cycle of cyclophosphamide and high-dose methyl prednisolone treatment and continued to receive interferon (IFN) alpha-2b therapy for 18 months. This treatment regimen resulted in an excellent response. In conclusion, LG may occur after the treatment of pediatric AML as a rare complication and IFN alpha-2b may be an effective treatment choice in these patients. PMID- 12571472 TI - Paroxysmal nocturnal hemoglobinuria arising from Fanconi anemia. AB - The progress of a female child with African type Fanconi anemia that evolves in time into paroxysmal nocturnal hemoglobinuria is described. Modern diagnostic methods are used to confirm this process. A discussion of possible mechanisms ensues. PMID- 12571473 TI - Refractory iron deficiency anemia as the primary clinical manifestation of celiac disease. AB - In the absence of dietary insufficiency, iron deficiency is usually caused by chronic blood loss or intestinal malabsorption. Celiac disease is one of the most common causes of intestinal malabsorption during childhood, and its association with insulin-dependent diabetes mellitus has been previously reported. Here the authors describe an otherwise asymptomatic diabetic adolescent boy with iron deficiency anemia that was not responsive to oral iron therapy. A diagnosis of celiac disease was made based on both anti-endomysial antibody titers and small intestinal biopsy. Institution of a gluten-free diet resulted in correction of the anemia. These observations emphasize the importance of considering a diagnosis of celiac disease in patients with nonresponsive iron deficiency anemia, particularly in the setting of insulin-dependent diabetes mellitus. PMID- 12571474 TI - Dapsone therapy for children with immune thrombocytopenic purpura. AB - Dapsone has been shown to be effective in treating adults with immune thrombocytopenic purpura (ITP). This retrospective review describes the authors' experience using dapsone in children with refractory, symptomatic ITP. Seven children were treated with dapsone. Dapsone was discontinued in two patients because of methemoglobinemia. In the remaining five patients, three achieved platelet counts of more than 100 x 10(3)/microL. Discontinuation resulted in a rapid decline in platelet counts in all three patients. Two of the three responded to a second round of treatment. Additional study of dapsone in children is warranted. Children receiving dapsone should be monitored for methemoglobinemia. PMID- 12571476 TI - Release of nucleated red blood cells early after cardiorespiratory arrest. PMID- 12571475 TI - Hypercalcemia in Langerhans cell histiocytosis: is it therapy-related? AB - Hypercalcemia is not known to be a common occurrence in patients with Langerhans cell histiocytosis (LCH). This manuscript describes an infant with multisystem LCH who developed an elevated serum calcium level while on therapy. Unlike other cases of hypercalcemia, this patient did not have evidence of bone involvement. Based on the apparent rarity of the condition and the clinical presentation, it is likely that hypercalcemia in LCH is a multifactorial process. The author presents evidence supporting the hypothesis that the treatment regimen is a major factor in the etiology of this metabolic abnormality and suggests that calcium levels be routinely measured during therapy. PMID- 12571477 TI - Complications of multilevel cervical corpectomies and reconstruction with titanium cages and anterior plating. AB - The ideal surgical treatment of multilevel cervical spondylosis remains unclear. This study analyzed the complications in using titanium cages and plating to reconstruct multilevel cervical corpectomies. This was a retrospective analysis of 21 consecutive patients who had multilevel cervical corpectomies and reconstruction with titanium cages and anterior plating. Sixteen had 2-level, one had 2.5-level, three had 3-level, and one had 3.5-level corpectomies. All had reconstruction with titanium cages and anterior plating. Thirty-three percent of the patients developed complications. Radiographs revealed bony consolidation in 95% of patients. Reconstructing multilevel cervical corpectomies with titanium cages and plating is associated with complications. Advantages include rigid immobilization and the avoidance of iliac crest bone graft harvesting. Major complications are largely the result of failures of the cage and plate construct, especially in patients with osteopenic bone. Supplemental posterior stabilization may be considered for cases with spasticity or greater than 2-level corpectomies with profound osteoporosis. PMID- 12571479 TI - Osteoporotic vertebral burst fractures with neurologic compromise. AB - A retrospective study was performed on the operative results following osteoporotic burst fractures with neurologic compromise. We sought to investigate the results of operative decompression and stabilization in patients with neurologic deficit as a result of an osteoporotic burst fractures. We examined the postoperative radiographic outcomes, level of disability, functional outcomes, and complications. Compression fractures of the anterior vertebral column secondary to osteoporosis and minimal trauma are a common clinical entity. These fractures are often effectively treated by nonoperative means. However, compressive failure of the middle vertebral column can lead to retropulsion of vertebral body fragments with significant canal compromise and neurologic injury. Treatment of these more severe injuries becomes more difficult and is less well established. Functional outcomes and disability from pain have not been examined. Previous reports on this subject have documented generally good results but have given few specific data regarding outcomes. We retrospectively report on a series of 10 patients, from 1995 to 1998, with osteoporotic burst fractures, which led to varying degrees of neurologic compromise. There were nine female and one male patient with a mean age of 76 years. Nine of the 10 patients presented more than 1 month following the onset of neurologic symptoms; 70% of the fractures occurred at the thoracolumbar junction (T11-L2). Mean loss of anterior column height was 59%, with significant kyphosis (mean 28 degrees) in nine of the 10 fractures. Mean canal compromise was 41%. At presentation, seven patients were Frankel grade D and three were Frankel grade C. All patients were treated operatively with decompression and arthrodesis. Mean time to follow-up was 16 months. Six of the 10 patients had improvement of their Frankel grade postoperatively and one deteriorated neurologically. Seven of the eight surviving patients completed the Oswestry questionnaire with a mean score of 44%, representing severe disability secondary to low back pain. The Physical Component score of the SF-36 was at or below the national mean for each patient. Complications were present in six of the eight surviving patients. Osteoporotic fractures are not benign. Careful evaluation for neurologic deterioration is warranted. Neurologic recovery occurred in six of the 10 patients; however, significant disability secondary to pain was common. PMID- 12571480 TI - Does donor site reconstruction following anterior cervical surgery diminish postoperative pain? AB - Many attempts have been made at donor site reconstruction to reduce postoperative pain following anterior cervical surgery. This study is a comparative analysis of the outcome of 46 patients undergoing single-level anterior corpectomy and fusion using iliac crest autograft performed by one surgeon (N.E.E.). Twenty-three patients had no donor site reconstruction, whereas the more recent 23 patients had iliac crest reconstruction using a MacroPore sheet (MacroPore, Inc., San Diego, CA) and Inductive Conductive Matrix (ICM, Sofamor Danek, Memphis, TN). Bodily Pain, assessed on the Short Form-36, obtained up to 12 months postoperatively, failed to demonstrate better pain relief following donor site reconstruction. Multiplanar CT studies obtained 6 months postoperatively documented 100% donor site fusion for the 23 reconstructed patients. Although iliac crest reconstruction failed to reduce Bodily Pain, it did result in 100% fusion. PMID- 12571481 TI - Harvesting bone graft from the posterior iliac crest by less traumatic, midline approach. AB - Complications of the donor site after the harvest of corticocancellous bone graft from the posterior iliac crest are very common. The most common are chronic donor site pain, tenderness, and sensory disturbances. This study investigates the results of the midline, lumbar fascia splitting approach for harvesting bone graft in lower lumbar spine fusion and compares them with the classic separate incision approach. A retrospective study of 107 patients compares two groups. The first group of 56 patients (35 males and 21 females with an average age of 41.8 years) had bone graft taken by splitting the two layers of the lumbar fascia down to their attachment to the iliac crest. The second group of 51 patients (29 males and 22 females with an average age of 43.7 years) had a separate incision over the iliac crest. In the first group, 82.1% had no tenderness, 8.9% mild, 7.1% moderate, and only 1.8% severe tenderness over the donor site. In the second group, 45.1% had no tenderness, 21.6% mild, 17.6% moderate, and 15.7% severe tenderness over the donor site. Five patients of the separate incision group (9.8%) had a lump in the donor site compared with none in the "same incision" group. Sensory disturbances over the donor site were found in 5.4% of the first group and in 21.6% of the second group. Harvesting bone graft from the posterior iliac crest for lower lumbar spine fusion through a midline, fascia splitting approach was found superior to the traditional, separate incision approach. PMID- 12571482 TI - Mixture ratios of local bone to artificial bone in lumbar posterolateral fusion. AB - There have been no reports on the effects of the mixture ratio of local bone graft to artificial bone on the outcome of intervertebral bone fusion. The purpose of this study was to determine whether the mixture ratio of local bone removed from laminae and spinous processes to apatite- and wollastonite containing glass ceramic (AWGC) granules affected fusion rates in spinal fusion. Posterolateral lumbar fusion at two levels without spinal instrumentation was performed in 35 patients who were diagnosed with lumbar spinal canal stenosis. The patients were randomly divided into three groups, each of which received a different mixture ratio of local bone to AWGC granules: 2:1, 1:1, and 1:2. The total mass of grafted bone was 20 g in all three groups. The fusion rate was approximately 80% in all three groups. Our results indicate that lumbar posterolateral fusion can be successfully achieved even when the mixture ratio of local bone graft to AWGC granules is 1:2. PMID- 12571483 TI - Radiologic findings of the lumbar spine in patients with rheumatoid arthritis, and a review of pathologic mechanisms. AB - We have analyzed the radiologic findings on the lumbar spine and the clinical symptoms in patients with rheumatoid arthritis (RA). A total of 106 patients who fulfilled the revised criteria of the American Rheumatism Association were subjected. All of the patients were asked to fill out a questionnaire about the existence of low back pain, leg pain, and leg numbness. Radiologic features of the lumbar spine, including scoliosis, spondylolisthesis, disc space narrowing, endplate erosion, osteophyte, and osteoporosis, were checked. Radiographs of the cervical spine were also taken. The clinical background of RA, such as mutilating disease or not, was assessed. Forty-two patients (40%) had the symptoms of low back pain. Abnormal radiologic findings in lumbar spine were detected in 57%. The prevalence of clinical symptoms tended to be higher in the patients with endplate erosion. Forty-two percent of the patients had both lumbar and cervical lesions. The prevalence of lumbar lesion was not high in the mutilating type of RA, except for facet erosion and severe osteoporosis. The patients with pulse steroid therapy revealed a higher prevalence of vertebral fracture. From these results, we concluded that lumbar lesions were frequently observed in patients with RA. The possibility of lumbar lesions as well as the lesions in the cervical spine and peripheral joints should be examined in patients with RA. PMID- 12571485 TI - Laminectomy contributes to cervical spine deformity demonstrated by holographic interferometry. AB - Multiple factors contribute to the pathogenesis of postlaminectomy deformity and instability of the cervical spine. The complex alterations in both static and dynamic biomechanics after laminectomy are incompletely defined. We sought to examine the role of the lamina in compressive load bearing across the vertebral body. Holographic interferometry was used to study the surface deformation of single axially loaded cervical vertebral bodies before and after hemilaminotomy, hemilaminectomy, and experimental acrylic laminar reconstruction. Our results showed that hemilaminotomy did not alter the surface deformation because of axial loading across the cervical vertebral body. However, gross alterations in surface deformation across the cervical vertebral body were consistently observed after hemilaminectomy. Experimental reconstruction of the laminar arch using acrylic restored the deformation pattern to the prelaminectomized baseline. Our results support a role for the lamina and the integrity of the laminar arch in axial load bearing across the cervical vertebral body. The altered axial load bearing may be a significant contributor to postlaminectomy deformity and instability. These findings offer an additional biomechanical advantage to minimal bony intervention for cervical spine pathology. PMID- 12571484 TI - Sagittal profile of the elderly. AB - Little is known about the natural history of spinal alignment as it ages into the eighth and ninth decades. Fifty asymptomatic volunteers 70-85 years of age (mean 76 years) without any history of spine pain, trauma, or deformity were radiographed in the standing lateral position, from C7 to the pelvis including the hips. Measurements included segmental angulations, kyphosis, lordosis, and C7 plumb line balance. In addition, measurements of sagittal pelvic balance were made (pelvic incidence, tilting, sacral slope, and S1 overhang). Average kyphosis was 52 degrees (range 29 degrees to 79 degrees); the average lordosis was -57 degrees (range -96 degrees to -20 degrees). The C7 plumb line on average fell 40 mm anterior to the posterosuperior corner of S1. The anterior positioning of C7 was also positively correlated with age and decreasing lordosis. This provides further data into the natural history of the aging spine. PMID- 12571486 TI - Finite element analysis of cervical spinal instability under physiologic loading. AB - The definition of cervical spinal instability has been a subject of considerable debate and has not been clearly established. Stability of the motion segment is provided by ligaments, facet joints, and disc, which restrict range of movement. Moreover, permanent damage to one of the stabilizing structures alters the roles of the other two. Although many studies have been conducted to investigate cervical injuries, to date there are only limited finite element investigations reported in the literature on the biomechanical response of the cervical spine in these respects. A comprehensive, geometric, nonlinear finite element model of the lower cervical spine has been successfully developed and validated under compression, anterior-posterior shear, and sagittal moments. Injury studies were done by varying each spinal component independently from the validated model. Seven analyses were conducted for each injury simulation (model without ligaments, model without facets, model without facets and ligaments, and model without disc nucleus). Results indicate that the role of the ligaments in resisting anterior and posterior shear and flexion and axial rotation moments is important. Under other physiologic loading (anterior-posterior shear, flexion extension, lateral bending, and axial rotation), the disc nucleus is responsible for the initial stiffness of the cervical spine. The results also highlight the importance of facets in resisting compression at higher loads, anterior shear, extension, lateral bending, and torsion. The results provide new insight through injury simulation into the role of the various spinal components in providing cervical spinal stability. These findings seem to correlate well with experimental results as well as with common clinical experience. PMID- 12571488 TI - Selection of fusion levels in scoliosis surgery. AB - Fusion levels are selected with the goal of having a balanced physiologic spine on the postoperative films. We have proposed an algorithm for selecting fusion levels based on the King-Moe scoliosis classification scheme and the flexibility of a compensatory curve. We propose the acronym TEAMS as a pneumonic. Curve type defines the region of the spine to be fused. The ends of the Cobb measurement around the region are the temporary endpoints. The endpoints are extended to avoid an apex due to the sagittal or frontal planes. The mobile disc is defined at the caudal end of the fusion area. Finally, the bending films, traction films, or intraoperative films are used to define the stable zone of Harrington that must contain the ends of the fusion mass. This scheme yields a simple, easily taught algorithm that results in reproducible endpoints for scoliosis fusion. PMID- 12571487 TI - Usefulness of functional outcome questionnaires in primary screening of spinal disease. AB - Back pain screening clinics are established to clinically screen patients with back pain for organic lumbar pathology. The aim of this study is to assess the relationship between clinical signs of organic pathology and the level of disability as measured by functional outcome scores. Sixty-nine patients who were found to have clinical signs of organic pathology and 69 age- and sex-matched patients from 512 patients who were found to have no signs of organic pathology in the same time period in the back pain screening clinic were selected. The Oswestry Disability, Short Form-36, and visual analogue (pain) scores between the two groups were statistically analyzed. We found a significant increase in the level of disability in the group with organic pathology; however, the variation in the scores makes them unspecific for use in screening of acute spinal pathology. We also found a strong relationship between psychologic disability and length of symptoms. PMID- 12571489 TI - Patterns of tumor spread and risk of fracture and epidural impingement in metastatic vertebrae. AB - Among patients with vertebral metastases, the identification of candidates for surgical stabilization has a limited basis in evidence. We retrospectively studied patterns of tumor spread (n = 756 vertebrae) and predictors of fracture and epidural impingement (n = 113 vertebrae) in infiltrated vertebrae with varying tumor histologies using sequential magnetic resonance images. Vertebral bodies were divided into 16 cells to map lesions. Fractured vertebrae were classified based on histology, level, fracture pattern, prefracture infiltration, and epidural impingement. Lesions were most often located within upper lumbar levels and the medial vertebral body. Fracture risk was greatest for upper lumbar (RR = 1.95; 95% CI: 1.12, 3.38) and undifferentiated tumors (RR = 7.36; 95% CI: 2.69, 20.12). A fourfold increase in fracture risk was noted in vertebrae with >80% body infiltration (HR = 4.5966; 95% CI: 1.66, 12.71). Symmetric fractures with fragments had the greatest risk of epidural impingement (p = 0.002). These findings have implications for management of patients with vertebral metastases. PMID- 12571490 TI - The effects of BMP-7 in a rat posterolateral intertransverse process fusion model. AB - The use of autologous bone grafting is an essential component in spine fusion because it is the key factor in achieving long-term stable arthrodesis between spinal motion segments. However, harvesting autologous iliac crest bone graft can be associated with significant morbidity and its supply is limited. Although no current substitute for autologous graft is available, multiple studies have already established the success of bone morphogenetic proteins (BMPs) in augmenting spinal fusion in models using larger animals. The purpose of our study was to evaluate the ability of BMP to augment a posterolateral intertransverse process single-level fusion in a rat. To our knowledge, this model has not been used to evaluate the effects of recombinant BMPs. A posterolateral intertransverse process fusion was attempted in white male Sprague-Dawley rats. The following are the four study groups: insoluble collagen bone matrix (ICBM) alone, 3 microg BMP-7 + 25 mg ICBM, 10 microg BMP-7 + 25 mg ICBM, and a sham group with no implanted material. The animals were killed on postoperative day 21 and were evaluated for signs of clinical and/or radiographic fusion. All of the rats in the 10 microg BMP-7 + 25 mg ICBM group demonstrated clinical fusion and had solid bilateral fusion masses on radiographs. None of the rats in the sham group, ICBM group, or 3 microg BMP-7 + 25 mg ICBM group fused clinically; however, the rats in the 3 microg BMP-7 + 25 mg ICBM group did show evidence of new bone formation. Our study demonstrates that a rat posterolateral intertransverse process fusion model is inexpensive and efficient and produces consistent results. It also shows that BMP can augment fusion in a rat and that dosing plays a role in successful fusion. This is consistent with results that have been studied in larger animal models. PMID- 12571491 TI - The sacroiliac joint: a potential cause of pain after lumbar fusion to the sacrum. AB - The sacroiliac joint (SIJ) can cause pain after lumbosacral fusion. Diagnosis requires >75% relief after local anesthetic SIJ injection. This study is a retrospective review of patients with low back pain after lumbosacral fusion who had SIJ injections. Percentage and duration of pain relief were noted. Results are as follows: there were 34 patients; 8 fused at L5-S1, 14 fused at L4-S1, and 12 had multilevel fusions. Twenty had >75% relief within 45 minutes, and 11 had prolonged relief. Six had relief >20% but <75%, and one had prolonged relief. Eight never improved. Eight had posterior iliac crest bone harvested, and there was no correlation between donor side and pain side. In 34 patients with low back pain after lumbosacral fusion, SIJ was the cause of pain in 32% and possibly the cause in 29%. This is the first detailed description of this problem. PMID- 12571492 TI - Coccygectomy for severe refractory sacrococcygeal joint pain. AB - Severe refractory coccydynia occasionally requires coccygectomy. Methods involved a retrospective chart review plus telephone follow-up of 15 patients who had coccygectomy. Pain was assessed by the numerical rating scale and function by Oswestry Low Back Disability Score. There were 13 (87%) patients (four men, nine women) available for follow-up at a mean of 43 months (range 19-74 months). Mean age was 45 years (range 24-72 years). All patients had coccygeal pain arising from the sacrococcygeal joint, but there were other lumbar disorders in 10 patients. There were two complications. The numerical rating scale improved from 7.3 to 3.6 (p < 0.001), and the Oswestry Low Back Disability Score improved from 55 to 36 (p < 0.001). Twelve patients would have the same surgery for the same result. Coccygectomy provides statistically and clinically significant improvement in patients with severe refractory sacrococcygeal joint pain. Many patients have other lumbar spine pathology. PMID- 12571493 TI - Spinal epidural abscess associated with IgG4 deficiency. AB - The objective is to present the possibility of an association between spinal epidural abscess and IgG deficiency. Spinal epidural abscess is a rare but potentially devastating condition. Review of the literature showed no reported acknowledgment about the relationship between spinal epidural abscess and IgG deficiency. This report discusses the case of a 16-year-old boy who developed progressive paraplegia within 24 hours. Clinical and neuroradiologic features are reported. Serum quantitative total IgG, IgA, and IgM concentrations were measured by nephelometry. Thoracic magnetic resonance imaging showed epidural abscess between T6 and T11 compressing the cord posteriorly. IgG subclasses (IgG ) were found abnormal. The possible importance of immunologic evaluation in the patients with spinal epidural abscess when no source of infection could be find is discussed.(4) (4) (4) PMID- 12571494 TI - Thoracolumbopelvic stabilization for the treatment of instability caused by recurrent myxopapillary ependymoma. AB - The authors report a patient with recurrent lumbosacral myxopapillary ependymoma, followed for more than 20 years, who presented with severe axial pain resulting from osteolytic destruction at the lumbosacral junction. Because the patient had a long history of paraplegia despite three previous incomplete tumor resections, we chose not to attempt a fourth resection. Moreover, because viable fixation points were not present within the sacrum and most of the lumbar spine, instrumented fusion was extended from T7 to the ilia using a modified Galveston L rod technique, which we believe to be unique in its application to this problem. This case demonstrates the long-term potential for instability from locally destructive myxopapillary ependymoma that is incompletely excised. We are not aware of any previous reports of lumbopelvic instability in association with myxopapillary ependymomas. PMID- 12571495 TI - Where's the evidence? Evidence-based orthopaedic trauma: a new section in the Journal. PMID- 12571496 TI - The effect of sacral fracture malreduction on the safe placement of iliosacral screws. AB - OBJECTIVES: To determine the effects of cranial displacement on the safe placement of iliosacral screws for zone II sacral fractures. DESIGN: Computer imaging and dimensional analysis of a human cadaveric sacral fracture model. SETTING: Cadaveric dissection, Orthopaedic Research Laboratories, Newark, New Jersey. MAIN OUTCOME MEASUREMENTS: Six cadaveric pelves with simulated zone II sacral fractures were imaged with computed tomography at controlled cranial displacements of 5, 10, 15, and 20 mm. The area of contact at the fracture site and volume of bone available for iliosacral screw placement was graphically measured using both two- and three-dimensional computer modeling. Areas of contact were also represented in terms of the maximal number of 7.0-mm screws that could be simultaneously implanted. RESULTS: Cross-sectional contact area was decreased by 30%, 56%, 81%, and 90% at 5, 10, 15 and 20 mm of displacement, respectively. Volume of bone was decreased by 21%, 25%, 26%, and 34% for 5, 10, 15 and 20 mm of displacement, respectively. In 50% of the specimens at 15 mm and 66% of the specimens with 20 mm displacement, two iliosacral screws could not be contained simultaneously within bone. In 17% of the specimens displaced 15 mm and 50% of the specimens displaced 20 mm, the cross-sectional area was insufficient to contain a single iliosacral screw. CONCLUSIONS: Although previous authors have accepted up to 15 mm of cranial displacement, the data demonstrate substantial compromise of available screw space with displacements greater than 1 cm. Fracture reduction is mandatory, as screw placement with residual displacement of 10 mm or more can endanger adjacent neural and vascular structures. PMID- 12571497 TI - The significance of anode location for stimulus-evoked electromyography during iliosacral screw placement. AB - OBJECTIVES: To determine the effect of anode location on the current threshold required to provoke an electromyograph response during stimulus-evoked electromyography for iliosacral screw placement. DESIGN: Prospective cohort. SETTING: Level I trauma center. PATIENTS: Nineteen consecutive patients with 23 unstable posterior pelvic ring injuries treated with iliosacral screws. INTERVENTION: Iliosacral screws were inserted percutaneously over guidewires. Twenty-seven screws were inserted, all into the first sacral vertebrae. The guidewire was used as the cathode for constant-current, stimulus-evoked electromyography for all data collection. Stimulus-evoked electromyographs were obtained with the guidewire at four different stations: at the sacroiliac joint (station I), at the first sacral neuroforamen (station II), in the body of the sacrum (station III), and when the iliosacral screw was in final position over the guidewire (station IV). MAIN OUTCOME MEASURE: Stimulus-evoked electromyographs were obtained with the anode at four different locations for each of the implant stations. Location A had the anode adjacent to the percutaneous insertion site of the guidewire, location B at the ipsilateral anterior superior iliac spine, location C at the midline, and location D at the contralateral anterior superior iliac spine. RESULTS: Moving the anode from midline (location C) toward the entry point of the guidewire increased the current threshold required to provoke an EMG response as much as 67.1% (p < 0.05). Moving the anode from midline to the contralateral anterior superior iliac spine decreased thresholds as much as 3.4% (p > 0.05). In one case, anode placement close to the guidewire insertion site (locations A and B) failed to identify a potentially dangerous implant because current thresholds were >8 mA. With the anode at the midline, current thresholds were <8 mA, indicating unsafe guidewire position leading to redirection of the guidewire. CONCLUSION: The physical location of the anode during stimulus-evoked electromyography monitoring for iliosacral screw placement significantly changes the current thresholds required to provoke an electromyograph response. Current thresholds required to stimulate nerves increase as the anode is moved toward the stimulating electrode. Anode placement ipsilateral to the stimulating electrode may provide a false indication of safe guidewire placement. We recommend anode location at or beyond the midline for stimulus-evoked electromyography monitoring during iliosacral screw placement. PMID- 12571498 TI - Optimal entry point for retrograde femoral nailing. AB - OBJECTIVE: The purpose of this study is to identify the optimum entry point for retrograde femoral nailing, defined as that point which will provide adequate fracture alignment while minimizing soft-tissue and articular cartilage injury. DESIGN: Cadaveric study. SETTING: Biomechanics laboratory. MAIN OUTCOME MEASURE: Anatomic relationships and fracture reduction. METHODS Eleven cadaveric femori with attached knee joints underwent retrograde femoral nailing with a Synthes femoral nail (Synthes, Paoli, PA, U.S.A.). After placement of the nail, the specimens underwent an osteotomy 3 inches proximal to the articular surface. Multiple entry points were tested to determine fracture alignment and extent of articular cartilage injury. Medial-lateral and anterior-posterior displacements, in addition to any soft-tissue or articular surface trauma, were recorded for these various points of entry. RESULTS: An entry point of 1.2 cm anterior to the femoral origin of the posterior cruciate ligament resulted in the least anterior posterior displacement of the femoral shaft following fracture. In the coronal plane, an entry point at the midpoint of the intercondylar sulcus was identified as minimizing the displacement following fracture. This ideal position allows for proper seating of the nail within the intercondylar sulcus, resulting in minimal damage to the articular cartilage and posterior cruciate ligament and minimal disruption of the patella femoral joint. CONCLUSION: Retrograde femoral nailing should be used cautiously in select patients, when conventional antegrade nailing cannot be used, due to the unavoidable injury to the knee articular surface associated with this technique. The optimum entry point of 1.2 cm anterior to the femoral posterior cruciate ligament origin and centered in the intercondylar sulcus provides the optimal balance of fracture reduction and knee joint sparing. It may be difficult to target this site with a percutaneous technique and may require direct visualization of the intercondylar sulcus for ideal nail placement. PMID- 12571499 TI - Displaced intra-articular calcaneal fractures: variables predicting late subtalar fusion. AB - OBJECTIVE: The goal of the current study was to analyze the prospective clinical outcome of patients who failed closed or open treatment of a displaced intra articular calcaneal fracture. This cohort of patients required a secondary subtalar fusion by distraction bone-block arthrodesis. DESIGN: Review of prospective, randomized trial database. SETTING: Four level I trauma centers. PATIENTS: Between April 1, 1991 and December 31, 1997, 424 patients with 471 displaced intra-articular calcaneal fractures were involved in a large, multicenter, randomized trial. Forty-four patients who required subtalar fusion following initial treatment of a displaced intra-articular calcaneal fracture were compared to the population of patients who did not require subtalar fusion. The variables compared between the two groups included Bohler angles, two computed tomography classification systems, and clinical scores including SF-36, visual analogue score, and oral analogue score. INTERVENTION: Subtalar distraction bone-block arthrodesis with tricortical bone graft was used in all 45 feet. MAIN OUTCOME MEASUREMENTS: The following were examined: x-ray fracture classification, specifically Bohler angles and Essex-Lopresti classification; computed tomography classification, specifically Sanders and Crosby; clinical scores, specifically validated visual analogue score, general health survey scores, oral analogue score, and other factors (i.e., patient demographics including age, sex, profession, smoking history, and Worker's Compensation Board involvement. RESULTS: Initial treatment of the 44 patients in our study was nonoperative in 37 (84%) patients and operative (open reduction and internal fixation) in 7 (16%) (1 patient had bilateral heel fractures). Patients requiring fusion differed demographically from those patients not requiring fusion. Mean age was 39 years in both the fusion and nonfusion group. The fusion group had 97% males, whereas the nonfusion group had 89% males. Sixty-four percent of the fusion patients were Worker's Compensation Board claims, whereas 35% of the nonfusion group were Worker's Compensation Board claims. Of those that required fusion, 77% were heavy laborers. On average, the fusion group had a Bohler angle 15 degrees less than the nonfusion group. Forty-six percent of the fusion patients were Sanders-type IV initial fractures. Logistic regression analysis revealed that the primary predictors of requiring fusion were Worker's Compensation Board status (odds ratio = 3.03, 95% confidence interval = 1.41 6.57), Sanders-type IV (odds ratio = 5.48, 95% confidence interval = 1.57-19.18), Bohler angle <0 degrees (odds ratio = 10.64-95% confidence interval = 1.33 85.17), and nonoperative initial treatment (odds ratio = 5.86-95% confidence interval = 2.33-14.67). CONCLUSION: These data suggest that the amount of initial injury involved with the calcaneal fracture is the primary prognostic determinant of long-term patient outcome. Bohler angle on presentation of <0 degrees was 10 times more likely to require a secondary subtalar fusion than a Bohler angle on presentation of >15 degrees. Sanders-type IV calcaneal fractures were 5.5 times more likely to be fused than a simple Sanders type II fracture. Worker's Compensation Board patients were three times more likely to be fused than non Worker's Compensation Board patients. Nonoperative care was six times more likely to lead to a late fusion as compared to open reduction and internal fixation treatment. Late fusion provided relief from pain and improved function as evidenced by an improvement in visual analogue score postsurgery. This study demonstrates that there is a distinct patient group with a displaced intra articular calcaneal fracture who are at high risk of subtalar fusion. These include male Worker's Compensation Board patients who participate in heavy labor work with a fracture pattern with Bohler angle less than 0 degrees. If their initial treatment was nonoperative, the likelihood of requiring late subtalar fusion was significantly increased. Initial open reductional open reduction and internal fixation of patients with displaced intra-articular calcaneal fracture minimized the likelihood that subtalar fusion would be required. PMID- 12571500 TI - Histomorphometry of distraction osteogenesis during experimental tibial lengthening. AB - OBJECTIVES: The aim of the current study was to analyze distraction osteogenesis histomorphometrically to investigate the amount of enchondral ossification adjacent to intramembranous ossification. STUDY DESIGN: Species-specific, longitudinal time study. METHODS Lengthening of 25% of the right tibia was performed in 24 beagle dogs by callus distraction after osteotomy and application of a ring fixator. Distraction was started at the fifth postoperative day with a distraction rate of 0.5 mm twice a day. Twelve dogs were killed at the end of the distraction phase of 25 days (group A) and the remaining 12 dogs after an additional consolidation period of 25 days (group B). The tibia was removed from the distracted right leg and from the left control side of each animal, and longitudinal sections were cut and stained with Pentachrome. In all tibiae, an area of 0.5 x 2.4 cm within the endosteal bone was evaluated histomorphometrically with a Merz grid, and the results were compared between distraction and control side as well as between the dogs of group A and group B. RESULTS: On the distraction side, the typical signs of an increased de novo bone formation with a significant increase of osteoblasts and osteoid in group A, as well as an additional significant increase of bone volume and trabecular thickness in group B, were found. In all distraction cases, islands of cartilage formation, which underwent enchondral ossification, were found, besides membranous ossification. The ratio between membranous and enchondral ossification was found to be five to one. CONCLUSION: Bone formation during distraction osteogenesis results from both membranous and enchondral ossification. PMID- 12571501 TI - Comparison of the mechanical properties of different tension band materials and suture techniques. AB - OBJECTIVES: This study determined the tensile properties of 18-gauge stainless steel wire, 5-mm woven polyester (Mersilene) tape, and multiple loop configurations of No. 5 braided polyester suture (Ethibond). DESIGN: Mechanical property testing. INTERVENTION: Single loops of stainless steel wire, Mersilene, and Ethibond were tested to determine their mechanical properties. Ethibond was tested with different numbers of loops and different knot configurations. MAIN OUTCOME MEASURES: Stiffness, load at failure, and elongation at failure. RESULTS: One loop of Mersilene and two loops of Ethibond had similar loads at failure, but the load at failure was significantly higher for stainless steel wire. Four loops of Ethibond withstood a similar failure load to stainless steel wire, but the failure load of the Ethibond suture was greater than the yield load of stainless steel wire. Stainless steel wire had a higher stiffness than both Ethibond and Mersilene. No difference was found in the failure load between different Ethibond knot configurations. The individually tied suture configuration resulted in a higher stiffness than the single-knot configuration. The elongation at failure was not statistically different among the different knot configurations and materials, with the exception of Mersilene tape. Mersilene tape demonstrated a significant increase in elongation at failure as compared to the other materials and knot configurations. CONCLUSION: It appears that multiple loops of Ethibond can substitute for stainless steel wire in situations where a compliant repair is suitable (support of a patellar tendon repair), but may not be satisfactory for rigid fixation (tension band fixation of a fracture). There appears to be no significant difference in strength but a small decrease in stiffness between tying multiple suture loops in one knot as opposed to individual knots. PMID- 12571502 TI - Postoperative upper extremity radiographs using the image intensifier: a simple adjunct to the "inverted" C-arm technique. AB - When the C-arm is used as a table in upper extremity surgery, postoperative plain radiographs can be obtained by placing the x-ray cassette directly on the image intensifier. This has many advantages, including more rapid performance than conventional techniques, a high-quality image, and no need for a recovery room radiograph. Additionally, there is no overlying splint material to obscure image detail, and the extremity can be positioned as desired by the operating surgeon. PMID- 12571503 TI - Apophyseal dislocation of the body of the sternum in a child: a case report. AB - A 4-year-old boy received a blow to the chest and sustained an apophyseal dislocation of the body of the sternum. The dislocation was treated by open reduction and internal fixation. This type of injury is so rare that no studies could be found. PMID- 12571504 TI - Combined Monteggia and Galeazzi fractures in a child: a case report and review of the literature. AB - We present an unusual case in which a combination of Monteggia and Galeazzi fractures occurred in the same forearm. The patient was a 10-year-old male who climbed up the pole of a basketball net, caught hold of the net, then lost his grip, and fell onto his right hand. On physical examination, a complete paralysis of the radial ulnar and median nerves was recognized. X-rays showed an olecranon fracture and lateral dislocation of the radial head in the elbow joint, a dorsal dislocation of the distal bone fragments due to a fracture of the distal third of the radius, and a palmar dislocation of the distal end of the ulna at the wrist joint. The injuries were diagnosed as a combination of a Bado type III Monteggia fracture and a palmar-type Galeazzi fracture of the same arm. Manual reduction and immobilization in a plaster cast were performed. Three years after the injury, both the distal and proximal radioulnar joints were maintained in the reduction position. Range of motion was reduced minimally in extension at the patient's elbow, and there was complete recovery of all three nerves. A combination of Monteggia and Galeazzi fractures in the same arm has been reported in only two pediatric patients worldwide and in eight cases total when adult patients are included, indicating that this is an extremely rare trauma. PMID- 12571505 TI - Successful open reduction of a 5-month-old hip dislocation associated with a femoral head fracture. AB - The long-term prognosis after open reduction of neglected posterior hip dislocations is poor; as such, primary arthroplasty is recommended by a number of authors. We present a patient with a 5-month-old posterior hip dislocation with concomitant paralysis of the sciatic nerve who had an open reduction. At a follow up of 3.5 years, the patient has normal function with no signs of arthrosis. Magnetic resonance imaging scans, however, revealed partial avascular necrosis of the femoral head. A discussion based on the literature shows the uniqueness of this case. PMID- 12571506 TI - Femoral neck/shaft fracture. PMID- 12571507 TI - Ipsilateral femoral neck and shaft fractures. PMID- 12571508 TI - Introduction: HIV/AIDS surveillance in a new era. PMID- 12571509 TI - HIV surveillance: a global perspective. AB - This article provides an overview of recommendations for HIV surveillance. Results of surveillance are used in practice to inform program decisions, judge the effectiveness of the national response, lobby for effective programs, and to provide accurate measures of trends and the absolute state of the epidemic. Recommended surveillance activities differ for different epidemic situations epidemics that are concentrated in defined groups with high-risk behavior, and epidemics that are well established among heterosexuals in the population at large. Surveillance systems in countries with different levels of the epidemic face major challenges, most of which revolve around identifying and obtaining information from representative samples of the at-risk populations. A brief examination of surveillance systems in Botswana and Vietnam illustrate how these challenges are being met in practice. While there is room for improvement, HIV surveillance systems in many developing countries are relatively robust, and are growing stronger all the time. In most countries, however, insufficient use is made of the information generated by these systems in terms of strengthening HIV prevention and care programs. PMID- 12571510 TI - Behavioral surveillance: current perspectives, and its role in catalyzing action. AB - In the age of 2nd generation surveillance, behavioral surveillance systems form one of the central components of national HIV monitoring systems. Different approaches to behavioral monitoring and the comparative role of behavioral surveillance are discussed, followed by an exploration of some of the practical issues that arise in the implementation of behavioral surveillance systems. This article concludes by stressing the importance of ensuring that the results of behavioral surveillance work are translated into actions that improve the national response to HIV. PMID- 12571511 TI - HIV/AIDS surveillance systems in Australia. AB - We review the HIV/AIDS reporting protocols in Australia, and describe the technical components of surveillance and its guiding principles, including a strict adherence to confidentiality at all levels. The majority of AIDS diagnoses in Australia have occurred in men who acquired HIV infection through male-to-male sex. The annual incidence of AIDS peaked in 1994, and then declined sharply over the subsequent 4 years. Advances in treatment effectiveness have focused attention on ensuring that people with HIV infection have access to optimal therapy, and emphasized the role of AIDS case reports as indicators of treatment failure. Analyses of the date of HIV diagnosis in people with AIDS have been used as indicators and predictors of undiagnosed HIV infection in the population. Australia's linkage of program funding to AIDS case counts provides an incentive for both health departments and community-based organizations to actively support AIDS surveillance activities. The challenge for surveillance is to maintain vigilance for shifts in HIV infection rates, or in behavior patterns that may be related to transmission. PMID- 12571512 TI - Combining data sources to monitor the HIV epidemic in Canada. AB - This article describes the methods, results and future perspectives of four information sources used to monitor the HIV epidemic in Canada: AIDS case surveillance, HIV case surveillance, HIV sentinel serosurveillance, and behavioral surveillance. Synthesizing data from these multiple sources provides a more comprehensive picture of the HIV epidemic than any one source alone could provide. In Canada, there has been a shift over time from an epidemic dominated by men who have sex with men to one where more than half of new infections are attributed to other groups, such as injection drug users and non-injecting heterosexuals. The available evidence also suggests increasing HIV infections among Aboriginal persons and among women. Surveillance data have been used in Canada to guide prevention and care programs and to formulate policy. In particular, these data have been used to support the development of an HIV testing program in pregnancy, to re-direct community work toward injection drug users and the young, and to demonstrate the effectiveness of new treatments for HIV. The main challenge now is to continue to improve the monitoring of the shifting HIV epidemic with more accurate data and to use the resulting information to inform appropriate prevention and care responses. PMID- 12571513 TI - HIV/AIDS surveillance in Denmark: the challenges ahead. AB - In Denmark, AIDS has been a mandatory reportable disease since 1983, and confirmed HIV infection has been the same since August 1990. The annual AIDS incidence increased initially and peaked in 1993 (4.6 per 100,000 inhabitants), then decreased to 1.2 per 100,000 inhabitants in 1998 and further to 0.9 in 2000. Most AIDS cases occur among men who have sex with men (MSM), representing 92% in 1980-1985 and 31% in 2000. Recently, AIDS incidence and mortality has decreased due to the new antiretroviral drugs. In 1995, 43 per million inhabitants died of AIDS, compared with 5 per million in 1998. HIV reporting in Denmark is anonymous. The annual number of new identified cases has been fairly stable at approximately 5.7 per 100,000 inhabitants. Immigrants represent 24% of identified HIV-infected persons and represent nearly 50% of all heterosexually acquired cases. Estimates show that HIV prevalence as of 2000 is 0.1% of the total population, distributed at 0.03% among heterosexuals and 4.8% among MSM. Estimated annual HIV incidence is around 5.6 per 100,000 inhabitants; three times higher among men than women, and as high as 220 per 100,000 among MSM. The spread of HIV is limited in Denmark but the prevalence is increasing due to the effect of antiretroviral therapy. This is a challenge to the existing HIV/AIDS surveillance and prevention strategy. PMID- 12571514 TI - Current situation and regional perspective on HIV/AIDS surveillance in europe. AB - AIDS reporting has been the principal means of monitoring the HIV/AIDS situation in Europe since 1984 [corrected]. HIV reporting was set up at the European level in 1999, although it has existed in most European countries since the 1980s. Anonymous individual data on AIDS and, if available, new HIV diagnoses, and data on HIV prevalence in various populations are reported from the 51 countries of the WHO European Region to EuroHIV, and aggregate data on HIV prevalence in various populations [corrected]. Data are presented after grouping the 51 countries into three geographic areas: the West, Center, and East. At end of 2000, in the West, AIDS incidence continued to decline except among those infected heterosexually; numbers of newly diagnosed HIV infections are relatively stable, but rising among heterosexually infected persons, many of whom originate from countries with generalized HIV epidemics. In the East, numbers of newly diagnosed HIV cases (mostly injection drug users) continue to rise steeply, particularly in the Russian Federation, Latvia, and Estonia. In the Center, levels of HIV and AIDS remain low. HIV reporting is becoming a central element of HIV/AIDS surveillance in Europe. Heterogeneity between countries in health care systems, HIV testing patterns and surveillance systems remain challenging for data standardization at the European level. Efforts should be made to use surveillance data for evaluating the effectiveness of HIV/AIDS prevention interventions. PMID- 12571515 TI - HIV/AIDS surveillance in Germany. AB - In Germany, since 1982, information on AIDS cases has been collected at the AIDS Center of the Robert Koch Institute. Since 1987, all laboratories performing HIV confirmatory testing have been required to report positive results anonymously. AIDS incidence peaked at about 2000 cases per year in 1993 and began to decline in 1995 following the widespread use of highly effective antiretroviral treatment. Current data indicate that the AIDS incidence has stabilized at a level of 750 cases per year since 1998. The number of newly diagnosed HIV infections has remained fairly stable at approximately 2000 to 2500 per year since 1993. Unlinked anonymous testing of dried blood spots from newborns is carried out in two federal states. The average prevalence of HIV seropositivity from 1993 to 1997 among women bearing children was 0.57 per 1000 in Berlin and 0.14 per 1000 in Lower Saxony. PMID- 12571516 TI - HIV/AIDS surveillance in Japan, 1984-2000. AB - The HIV/AIDS surveillance system in Japan, which began collecting data on the number of AIDS patients in 1984 and the number of HIV-infected persons in 1987, has played an important role in monitoring the trend and magnitude of Japan's HIV/AIDS epidemic and its distribution across various population subgroups. However, the system lacks any personal identifiers, making it impossible to eliminate duplication or to track cases for disease progression. It also does not permit the identification of the residence of HIV-infected persons because the residence of only the reporting physician is documented under the New Infectious Diseases Control Law, effective since April 1, 1999. The number of people with HIV/AIDS in Japan continues to grow. Among youth, sexually transmitted diseases, induced abortion, and sexual activities have shown a marked increase since the mid-1990s. Behavioral risk of infection for both injection drug users (IDUs) and men who have sex with men (MSM) remains alarmingly high. Accurate monitoring of infection rates is critical to the planning and evaluation of treatment, care and prevention programs. Japan should restructure its HIV/AIDS surveillance system to more accurately monitor the HIV/AIDS epidemic and related risk behaviors. PMID- 12571517 TI - Thailand report: HIV/AIDS surveillance 1998. AB - Patients identified as having symptomatic HIV infection or AIDS have been reported to the Division of Epidemiology (DOE), in the Thai Ministry of Public Health (MOPH) since 1984. In addition, since 1989, the DOE and the Royal Thai Army have conducted annual serosurveillance of HIV in particular populations. Sexual behavior surveys have been conducted annually since 1995. HIV prevalence among direct commercial sex workers (CSWs) peaked in 1994 (33%) but has declined to 22%. Prevalence among male CSWs reached 18% in 1994 but has not changed since. HIV prevalence among blood donors was 1% in 1992, but has declined to 0.5%. HIV prevalence among pregnant women peaked in 1995 (2.3%) and has since declined to 1.5%. HIV prevalence among 21-year-old male conscripts peaked in 1993 (4%), but declined to 1.9% in 1998. Prevalence among IDUs has remained between 30%-40% since the late 1980's. The number of reported AIDS cases was 128,606 by October of 1999, and has increased yearly. Nonetheless this figure is likely unreliable due to significant underreporting. Thailand's surveillance program appears to be effective in increasing public knowledge and information about the epidemic, although it faces growing problems of good data management, interpretation, timely distribution of information, and investigation of abnormal events. PMID- 12571518 TI - HIV/AIDS surveillance in the United States, 1981-2001. AB - We review the HIV/AIDS reporting system, including the legal basis for reporting, the methods and infrastructure for reporting, evaluation of the completeness and quality of the data, and analysis and dissemination of reports. Other information systems (e.g., seroprevalence surveys and behavioral surveys) that collect useful information for HIV prevention and care programs are also described. Multiple data collections systems are needed to monitor the HIV/AIDS epidemic in the United States and to collect the information needed to plan, implement, and evaluate prevention and care programs. PMID- 12571519 TI - Taking informed consent seriously in global HIV vaccine research. PMID- 12571520 TI - Population survey of CCR5 delta32, CCR5 m303, CCR2b 64I, and SDF1 3'A allele frequencies in indigenous Chinese healthy individuals, and in HIV-1-infected and HIV-1-uninfected individuals in HIV-1 risk groups. AB - The aim of this study is to determine in indigenous Chinese ethnic groups the frequencies of the chemokine (SDF1 3'A) and chemokine receptors (CCR5 delta32, CCR5 m303, and CCR2b 64I) HIV-1/AIDS restriction alleles. The study includes two cohorts; the first comprised 3165 indigenous healthy subjects representing eight ethnic groups: Han (n = 1406), Uygur (n = 316), Mongolia (n = 134), Hui (n = 386), Tibetan (n = 330), Zhuang (n = 378), Dai (n = 101), and Jingbo (n =114). The second cohort consisted of 330 HIV-1-infected (86 subjects infected by sexual transmission and 198 subjects infected by HIV-1-contaminated blood or by sharing injection equipment; the remaining 46 subjects said nothing about HIV-1 transmission) and 474 HIV-1-uninfected Han Chinese belonging to one of two HIV-1 high-risk groups: intravenous drug users (n = 215) and individuals with sexually transmitted diseases (n = 259). Genotypes for the four genes were obtained using PCR (CCR5 delta32) or PCR-restriction fragment length polymorphism. Randomly selected amplified PCR products were further confirmed by direct DNA sequencing. The variant allele frequencies were determined to be 0% to 3.48% for CCR5 delta32, 0% for CCR5 m303, 16.23% to 28.79% for CCR2b 64I, and 17.70% to 27.76% for SDF1 3'A in Chinese healthy individuals from eight ethnic groups. These findings show that allele frequencies differ among the eight Chinese ethnic groups for CCR5 delta32, CCR2b 64I, and SDF1 3'A and that the CCR5 m303 and CCR5 delta32 mutant alleles were absent or infrequent in Chinese, which may be helpful for studies of specific anti-HIV-1 vaccine trials and coreceptor inhibitor drug targets in Chinese populations. Furthermore, we observed no significant differences in allele or genotypic frequencies between HIV-1-infected and HIV-1 uninfected groups from the Han ethnic group. Our finding is the first reporting that there is likely no effect of the examined polymorphisms in our study on HIV 1 transmission in the Chinese Han population, However, the genetic effects of these and other AIDS-modifying polymorphisms on the pathogenesis and clinical outcome of HIV-1/AIDS diseases is under investigation in Chinese populations. PMID- 12571521 TI - Distribution of the CCR5 gene 32-base pair deletion and CCR5 expression in Chinese minorities. AB - China has an ethnically diverse population. Genetic differences may contribute to disparities in the efficiency of HIV transmission. To further characterize this risk, we examined the HIV-related genetic diversity in the predominant Han Chinese and in six minority groups. We searched for the delta32-CCR5 mutation, a common cause of relative HIV resistance in the white population. In addition, CCR5 receptor expression was measured. Blood samples were obtained from adults belonging to the Han, Meng, Zang, Weiwuer, Zhuang, Yi, and Dai ethnic groups. Polymerase chain reaction analysis was performed on genomic DNA samples. Surface expression of CCR5 on peripheral blood mononuclear cells was measured by flow cytometry. One-way ANOVA was used to determine mean statistical differences. Samples from 10 members of each minority were examined. A delta32-CCR5 heterozygote phenotype was detected in one Weiwuer subject, but no mutations were found in the other 69 subjects studied. The mean CCR5 expression of cells harvested from the Dai minority was greater than that of cells from all other minorities studied, for both CD3+CCR5+ and CD4+CCR5+ sets (p < .01, one-way ANOVA). The delta32-CCR5 mutation seems to be rare in most Han Chinese and the minority populations studied. CCR5 expression appears to be greater in the Dai minority than in the other minorities investigated. The mechanism for this increased expression requires further study. PMID- 12571522 TI - Persistent leukocyturia and loss of renal function in a prospectively monitored cohort of HIV-infected patients treated with indinavir. AB - Symptomatic nephrotoxicity is a well-known complication of indinavir treatment. However, little is known about the relevance of other abnormalities, such as leukocyturia during use of indinavir. We determined the prevalence, risk factors, and consequences of persistent leukocyturia in a prospectively monitored cohort of indinavir users in three adult outpatient clinics. Patients were monitored for nephrotoxicity at regular visits (every 3 months) between August 1998 and September 2000. Monitoring involved urine dipstick analysis and microscopy for pH, erythrocytes, leukocytes, and indinavir crystals. The urine albumin concentration/creatinine concentration ratio and serum creatinine and indinavir plasma concentrations were measured, and urinary tract infection was excluded. Urologic symptoms were retrieved from medical records. Of 184 patients with at least one assessment, 35% had leukocyturia (i.e., >75 cells/microL) at least once during the study period, which coincided with mild increase in the serum albumin level, erythrocyturia, and crystalluria. Thirty-two (24%) of 134 patients with two or more assessments had persistent leukocyturia (i.e., on two or more occasions). Risk factors were indinavir plasma concentration of >9 mg/L, urine pH of >5.7, and crystalluria. Persistent leukocyturia was associated with a gradual loss of renal function but not with urologic symptoms. The data show that leukocyturia is a frequent finding and emphasize the need for monitoring renal function during indinavir treatment, even in the absence of urologic symptoms. PMID- 12571523 TI - Review of medical encounters in the 5 years before a diagnosis of HIV-1 infection: implications for early detection. AB - Early detection of HIV infection improves prognosis and reduces transmission, but 30%-40% of cases are diagnosed late. A comprehensive and systematic review of medical encounters before diagnosis has not been done. This study reviews 5 years of medical encounters before the diagnosis of HIV infection in members of a large managed care organization where access to care is reasonably good. Patient characteristics, HIV risk factors, and clinical events preceding diagnosis were examined and tested for association with late diagnosis (CD4 cell count of <200/microL at diagnosis). Of 440 HIV-infected patients, 62% had CD4 cell counts of <350/microL, 43% had CD4 cell counts of <200/microL, and 18% had CD4 cell counts of <50/microL at diagnosis. Twenty-six percent of all patients had risks documented >1 year before diagnosis. Only 22% of patients had one of eight clinical indicators suggested in the literature as reasons to test for HIV >1 year before diagnosis. In multiple logistic regression, older age, male sex, race, risk group, no prior HIV testing, physician-initiated testing, and having any of eight clinical indicators before diagnosis were each associated with late diagnosis (p 90% at 29 days of age and thereafter; specificity was 100% in all periods, except at 29 to 60 days of age, when specificity was 93%. Neither sensitivity nor specificity was significantly affected by maternal or infant zidovudine (ZDV) treatment, even though infant viral loads were lower during the first 6 weeks in infants who received perinatal ZDV prophylaxis ( p=.005). Paired analysis of DNA and RNA measurements revealed no advantage for either test. Quantitative RNA testing can be used for diagnosis in HIV-exposed infants, recognizing the chance for a false-positive test result. It may be most useful as a confirmatory test in infants with another positive diagnostic test result. PMID- 12571530 TI - Support among persons infected with HIV for routine health department contact for HIV partner notification. AB - Public health partner notification (PN) services are provided inconsistently to persons diagnosed with HIV/AIDS in the United States, and some community groups representing persons with HIV/AIDS have opposed widespread application of PN. We surveyed persons with HIV recently reported to our health department and a random sample of HIV-infected persons attending an HIV/AIDS clinic. A total of 95 persons, of whom 76 (80%) were men who have sex with men, completed an anonymous self-administered questionnaire. Eighty-four percent of participants believed the health department should routinely offer everyone diagnosed with HIV help in notifying their partners; 79% indicated they would be somewhat or very likely to provide information to a doctor, case worker, or health department employee for purposes of PN; and 20% indicated they wanted help in notifying a recent sex partner. Seventy-eight percent of participants believed the health department should contact all HIV-infected persons after diagnosis to help them access medical care and social services, and 68% wanted the health department to contact them about the availability of medical or social services. In contrast to common public perceptions, these results suggest that most persons with HIV support health departments routinely contacting people after HIV diagnosis and that many want assistance with PN. PMID- 12571531 TI - Development of proteinuria or elevated serum creatinine and mortality in HIV infected women. AB - BACKGROUND: Data on the incidence and prognostic significance of renal dysfunction in HIV disease are limited. OBJECTIVE: To determine the incidence of proteinuria and elevated serum creatinine in HIV-positive and HIV-negative women and to determine whether these abnormalities are predictors of mortality or associated with causes of death listed on the death certificate in HIV-positive women. DESIGN: The incidence of proteinuria or elevated serum creatinine and mortality was assessed in a cohort of 885 HIV-positive women and 425 at-risk HIV negative women. SETTING: Women from the general community or HIV care clinics in four urban locations in the United States. OUTCOME MEASURES: Creatinine of >or=1.4 mg/dL, proteinuria 2 or more, or both. Deaths confirmed by a death certificate (92%) or medical record/community report (8%). RESULTS: At baseline, 64 (7.2%) HIV-positive women and 10 (2.4%) HIV-negative women had proteinuria or elevated creatinine. An additional 128 (14%) HIV-positive women and 18 (4%) HIV negative women developed these abnormalities over the next (mean) 21 months. Relative hazards of mortality were significantly increased (adjusted relative hazard = 2.5; 95% confidence interval: 1.9-3.3), and there were more renal causes recorded on death certificates (24/92 (26%) vs. 3/127 (2.7%), p<.0001) in HIV infected women with, compared with those without these renal abnormalities. CONCLUSIONS: Proteinuria, elevated serum creatinine, or both frequently occurred in these HIV-infected women. These renal abnormalities in HIV-infected women are associated with an increased risk of death after controlling for other risk factors and with an increased likelihood of having renal causes listed on the death certificate. The recognition and management of proteinuria and elevated serum creatinine should be a priority for HIV-infected persons. PMID- 12571532 TI - C-reactive protein is an independent predictor of mortality in women with HIV-1 infection. AB - The relationship of C-reactive protein (CRP) to mortality was assessed in 209 HIV 1-infected women after adjusting for age, body mass index (BMI), serum albumin, CD4 cell lymphocyte count, and HIV-1 RNA. During the follow-up period of up to 5 years (median = 45 months) there were 49 deaths. CRP at study enrollment was measured using a low sensitivity assay. CRP levels were only weakly correlated (Pearson correlation coefficient r < .2) with other predictors of mortality. CRP was a powerful predictor of mortality (p < .01) after adjusting for age, BMI, serum albumin, CD4 cell lymphocytes, and HIV-1 RNA. The relative hazard associated with an elevated CRP level, independent of the covariates noted above, varied from 3.4- to 13.6-fold depending on how CRP values were grouped. CRP may be a useful and inexpensive predictor of HIV disease mortality in women. PMID- 12571533 TI - HIV-1 infection among civilian applicants for US military service, 1985 to 2000: epidemiology and geography. AB - OBJECTIVES: This study examined demographic and geographic correlates of HIV-1 prevalence among civilian applicants for US military service. METHODS: HIV-1 test results and demographic and geographic data were available for 5.3 million applicants. RESULTS: Between October 1985 and December 2000, a total of 5,340,694 individuals applied to join one of the armed service branches of the US military. Overall, HIV-1 prevalence was 0.80 per 1000 applicants (95% CI: 0.78-0.82), with 4276 applicants testing positive for HIV-1 infection. Prevalence declined over the 16-year period from a high of 2.89 per 1000 applicants in 1985 to 0.36 per 1000 applicants in 2000. The majority of applicants (82.7%) were male, and the majority of HIV-1 cases (89.4%) occurred in men. HIV-1 prevalence was higher among African Americans (2.47/1000) and Hispanics (0.90/1000) than among white applicants (0.36/1000). HIV-1 prevalence was lowest in the West North Central region of the United States (0.33/1000) and highest in the Middle Atlantic region (1.61/1000) and Puerto Rico (3.56/1000). CONCLUSIONS: Civilian applicants for US military service comprise a unique cohort for following trends in the evolving HIV-1 epidemic. PMID- 12571534 TI - Risk factors for pneumonia in urban-dwelling HIV-infected women: a case-control study in Nairobi, Kenya. AB - In sub-Saharan Africa, respiratory tract infections (RTI) are the leading cause of serious morbidity and mortality in HIV-infected persons. This study sought to investigate demographic, socioeconomic, and environmental risk factors for pneumonia in a cohort of HIV-infected women. The authors performed a nested case control study in a cohort of HIV-1-infected adults followed in Nairobi, Kenya. Thirty-nine women who developed pneumonia during the follow-up period were selected as cases, and 66 women who did not develop pneumonia were randomly chosen to serve as control subjects. A questionnaire was administered in subjects' homes that assessed demographics, home environment, and socioeconomic status. Women were followed in the cohort for a median of 36.8 months (range, 27.3-39.3). Adjusting for length of follow-up period, factors associated with lower socioeconomic status (lower monthly spending [OR = 3.2; 95% CI, 1.2-8.4 per 10,000 Kenyan shilling decrease], having no savings [OR = 4.1; 95% CI, 1.4-11.9], less sturdy home construction material such as mud or cement walls [OR = 2.6; 95% CI, 1.1-5.9] or dirt floors [OR = 2.8; 95% CI, 1.0-7.6], and lack of a window in the home [OR = 5.5; 95% CI, 0.9-32.2]) and being widowed (OR = 4.3; 95% CI, 1.2 15.1) or single (OR = 3.3; 95% CI, 1.0-11.2) were associated with an increased risk of pneumonia. In multivariate analysis, widowed (AOR = 5.9; 95% CI, 1.3 26.3), single (AOR = 7.7; 95% CI, 1.6-36.4), and divorced (AOR = 4.5; 95% CI, 1.0 20.1) women, those without savings (AOR = 3.7; 95% CI, 1.2-11.7), and those living in more crowded and contagious conditions (AOR = 1.5; 95% CI, 1.1-2.1) remained at increased risk of pneumonia. If confirmed by prospective investigation, these findings could help identify persons and subpopulations of HIV-infected women with the greatest risk of pneumonia. PMID- 12571535 TI - Readiness for cessation of drug use among recent attenders and nonattenders of a needle exchange program. AB - Needle exchange programs (NEPs) represent a bridge to drug abuse treatment. NEP attenders tend to have more severe drug problems, however, and may be less ready to reduce their drug use than other drug users. This study investigated the relationship between NEP attendance and readiness for cessation of drug use. Since the period from 1988 through 1989, a community-based sample of injection drug users (IDUs) in Baltimore has undergone semiannual interview-administered questionnaires and HIV testing. A total of 288 IDUs completed a questionnaire on readiness for cessation of drug use. Readiness for drug use cessation was assessed from a 28-item validated scale of problem drug use and intention to quit, based on the "stages of change" model. Logistic regression was used to determine factors associated with readiness for cessation of drug use, including sociodemographics, drug use behaviors, and NEP attendance. Thirty percent of respondents attended the NEP in the past month. Stage of change in readiness for cessation of drug use did not differ between NEP attenders and nonattenders (OR = 0.9; 95% CI: 0.5-1.6). Similar proportions of persons recently attending and not attending the NEP were classified as ready to stop drug use (about 30%). In multivariate analysis, readiness for cessation of drug use was associated with speedball injection and previous enrollment in drug treatment but not with NEP attendance. NEP attenders, although exhibiting characteristics consistent with more severe drug dependence, were as motivated for cessation of drug use as were nonattenders. These findings suggest that formal collaboration between NEPs and drug treatment programs could increase the proportion of IDUs in treatment. PMID- 12571536 TI - The efficacy of lopinavir in individuals experiencing protease inhibitor failure. PMID- 12571537 TI - Once-daily saquinavir-sgc plus low-dose ritonavir (1200/100 mg) in combination with efavirenz: pharmacokinetics and efficacy in HIV-infected patients with prior antiretroviral therapy. PMID- 12571538 TI - Teriparatide (forteo) for osteoporsis. PMID- 12571539 TI - Atomoxetine (strattera) for ADHD. PMID- 12571540 TI - Augmentin XR. PMID- 12571541 TI - Oxaliplatin (Eloxatin) for advanced colon cancer. PMID- 12571542 TI - Targeted molecular therapy for cancer: the application of STI571 to gastrointestinal stromal tumor. PMID- 12571543 TI - Can treatment with angiotensin-converting enzyme inhibitors in elderly patients with moderate to severe chronic heart failure be improved by a nurse-monitored structured care program? A randomized controlled trial. AB - OBJECTIVE: The purpose of this study was to examine whether a nurse-monitored structured care program resulted in a more effective use of angiotensin converting enzyme (ACE) inhibitors in elderly patients compared with standard care in patients with chronic heart failure (CHF). METHODS: Hospitalized patients were screened to identify individuals with CHF, age more than 65 years, New York Heart Association classification III to IV, and no contraindications to ACE inhibitor treatment. One hundred forty-five patients were randomized to a nurse monitored structured care program that included uptitration of enalapril to a target dose of 10 mg twice a day or to standard care. Six-month follow-up data were collected. RESULTS: The mean age of the randomized patients was 81 years. Although the proportion of patients treated with an ACE inhibitor did not differ between structured care (70%) and standard care (64%), the number of patients with the target ACE inhibitor dose was significantly higher in the structured care group (26% versus 11% in the standard care group; P <.018). Treatment had to be discontinued in 26% of the patients because of adverse effects. CONCLUSION: The patients in this study were older than in previous intervention studies and had considerable comorbidity and reduced tolerance for ACE inhibitors. ACE inhibitor treatment was underused but improved with the structured care program, although achieved treatment levels were below those in the large intervention trials in patients with CHF. PMID- 12571544 TI - Cardiovascular autonomic function in healthy adolescents. AB - OBJECTIVE: The purpose of this study was to determine effects of age, sex, race, body mass index, and Tanner's stage on short-term evoked cardiovascular autonomic tests (ie, Valsalva ratio and change in heart rate with deep breathing) and 24 hour heart rate variability (HRV) in a sample of healthy adolescents, as well as to identify normative indices of both short-term evoked and 24-hour HRV in this age group. DESIGN: A descriptive, correlational design was used. SETTING: Study took place in a university hospital in a health science center located in the mid South. SUBJECTS: Participants included 75 healthy adolescents: mean age was 15.0 +/- 1.6 years, 14 were African American, 61 were white, 49 were girls, and 26 were boys. OUTCOME MEASURES: Study measures included the Valsalva ratio, change in heart rate with deep breathing, and 24-hour HRV with power spectral analysis with Holter monitoring. RESULTS: Major significant findings included lower values of 24-hour HRV measures for girls and African American adolescents (P <.05). Indices for normal ranges of both the short-term evoked and 24-hour HRV measures were computed with 95% confidence intervals. CONCLUSIONS: Few published studies address cardiac autonomic function, including 24-hour HRV, in adolescents. Most studies reporting actual normative control values of HRV for youth typically have not addressed sex or racial differences. Our study included the largest number of adolescents to date in the reported literature and demonstrated the importance of considering sex and race variation in interpreting test results. The availability of state-of-the-art technology for obtaining HRV data allows for the early identification of subclinical cardiac autonomic changes in youth who have predispositions for cardiac complications, such as those with diabetes, congenital heart disease, or obesity. PMID- 12571546 TI - Qualitative analysis of living with heart failure. AB - BACKGROUND: The incidence of heart failure has been described as epidemic in proportion. Although literature abounds surrounding issues of epidemiology, pathophysiology, and treatment issues for those with heart failure, little is known about the day-to-day experiences of adults living with this chronic illness. OBJECTIVES: The purpose of this study was to describe and analyze the experience of adults who are living with heart failure. The specific aims were to describe the problems, challenges, and processes of living with heart failure, to identify strategies and tactics adults use to live with heart failure, and to provide a model that articulates what the participants described. METHODS: Principles of naturalistic inquiry were used to describe problems, challenges, and processes of living with heart failure. Six women and 5 men participated in in-depth interviews that lasted from 40 minutes to 100 minutes. Data were collected and analyzed with the constant, comparative method. RESULTS: Participants' use of language referring to wind and water in describing their heart failure symptoms led to discovery of navigating and aspects of navigational science as metaphors for living with heart failure. The 3 main categories of the resulting model were called experiencing turbulence, navigating, and finding safe harbor. CONCLUSIONS: Use of the model for living with heart failure may provide for creation of interventions for adults to improve their ability to manage their own care in the face of this potentially devastating illness. PMID- 12571545 TI - Patient anxiety while on a waiting list for coronary artery bypass surgery: a qualitative and quantitative analysis. AB - OBJECTIVE: The purpose of this study was to describe the nature and intensity of anxiety felt by patients awaiting coronary artery bypass surgery. DESIGN: A prospective, cross-sectional study design was used, with a qualitative interview and State Trait Anxiety Inventory. SETTING: Study took place in 2 large tertiary referral hospitals in Northern Ireland. PARTICIPANTS: Seventy patients were randomly selected within 4 weeks of their referral for cardiac surgery. RESULTS: Participants in this study cited 5 main sources of anxiety: chest pain, uncertainty, fear of the operation, physical incapacity, and dissatisfaction with the care offered to them. The State Trait Anxiety Inventory scores of this sample were high at all stages of data collection. There was a statistically significant relationship (P or = 80 and IgA > or = 40 were defined as elevated antibody titres. Epstein-Barr antibodies were measured in order to investigate whether a possible association exists between hypertension and other, similarly widespread in the general population, intracellular microorganisms. All participants underwent casual blood pressure (BP) readings and 24 h ABPM. Subjects having mean 24 h systolic/diastolic ambulatory BP>125/80 mmHg, with or without antihypertensive medication were defined as hypertensives. Controls were free of any history or clinical evidence of hypertension, cardiovascular or pulmonary disease. Of the total participants, 77 hypertensives (52.7%) and 10 normotensives (18.5%) had IgA titres > or = 40 (crosstabs P < 0.000), whereas 76 hypertensives (52.1%) and 15 normotensives (27.8%) had IgG titres > or = 80, (crosstabs P < 0.002). No difference was found in Epstein-Barr antibodies, between hypertensives and normotensives. In conclusion, C. pneumoniae, but not Epstein-Barr, antibody levels were found significantly higher in sustained hypertensives, suggesting high frequency of chronic C. pneumoniae, infections in this specific group of patients. PMID- 12571614 TI - Dietary risk markers that contribute to the aetiology of hypertension in black South African children: the THUSA BANA study. AB - Although clinical hypertension occurs less frequently in children than in adults, ample evidence supports the concept that the roots of essential hypertension extend back to childhood. Since little is available in the literature on causal dietary factors of hypertension in children, this study hypothesised that certain dietary factors can be identified as risk markers that might contribute to the aetiology of hypertension in black children. Children aged 10-15 years were randomly selected from 30 schools in the North West Province from 2000 to 2001. These children comprised 321 black males and 373 females from rural to urbanised communities, of which 40 male and 79 female subjects were identified with high normal to hypertensive blood pressure. Blood pressure was measured with a Finapres apparatus and data were analysed with the Fast Modelflo software program to provide systolic, diastolic and mean blood pressure. A 24-h dietary recall questionnaire and weight and height measurements were taken. In a stepwise regression analysis, the following variables were significantly associated (P < or = 0.05) with blood pressure parameters of hypertensive males: biotin, folic acid, pantothenic acid, zinc and magnesium. Energy, biotin and vitamin A intakes were significantly associated with blood pressure parameters of hypertensive females. No significant dietary markers were indicated for any of the normotensive groups. Dietary intakes of all of these nutrients were well below the dietary reference intakes. In conclusion, the dietary results coupled with the cardiovascular parameters of this study identified folic acid and biotin as risk markers that could contribute to the aetiology of hypertension in black persons. The low intakes of these nutrients, among others, is a matter of serious concern, as is the increasing tendency towards urbanisation. PMID- 12571615 TI - Hypertension in diabetes: trends in clinical control in repeated large-scale national surveys from Sweden. AB - Hypertension in diabetes is an important and treatable cardiovascular risk factor. Treatment targets from guidelines cannot always be achieved in everyday clinical practice. It is therefore of great importance to monitor trends in hypertension control in defined populations. Patients with type I diabetes (range 6685-10,100; treated hypertension 21-29%) or with type II diabetes (range 15,935 22,605; treated hypertension 47-56%) were included in four national samples between 1996 and 1999. This screening was part of the procedures for the National Diabetes Register in Sweden, which monitors trends in clinical practice and risk factors for patients with diabetes, recruited both in primary health care and at the hospital level. A favourable trend in mean and median blood pressure levels was noticed during the 4-year study period, based either on data from repeated surveys or on repeated measures in the same individual, both for type I diabetes (mean: -2/-2 mmHg; P < 0.01) and for type II diabetes (mean: -5/-3 mmHg; P < 0.001). Correspondingly, the proportion of hypertensive patients in acceptable control of blood pressure (< or =140/85 mmHg) increased (P < 0.001) both in type I diabetes (52.0-57.9%) and in type II diabetes (22.4-33.3%). It was concluded that hypertension is a widespread cardiovascular risk factor in patients with diabetes, especially systolic hypertension. A trend for a better systolic blood pressure control during the late 1990s in hypertensive patients with type II diabetes in Sweden could translate into substantial (estimated) clinical benefits in cardiovascular and diabetes-related morbidity. The National Diabetes Register makes a quality assessment of the hypertension treatment possible. PMID- 12571616 TI - Blood pressure measurement in an ambulatory setting: concordance between physician and patient self-measurement. AB - The aim of this study was to determine concordance between physician and patient blood pressure (BP) measurements in an ambulatory setting. A diagnostic intervention cross-sectional study using a convenience sample was employed. A total of 106 hypertensive patients were included in the study. Patients who were unable to perform their self-measurement or those with cardiac arrhythmia were excluded. BP was determined nine times in each subject in the medical office in a randomised order: BP was taken three times by the physician using a mercury sphygmomanometer (SPH-Hg), three times by the physician using a validated, automated oscillometer (Omron HEM 705 CP), and three times by the patient himself with the same device. The intraclass correlation coefficient was calculated. In all, 59 women and 47 men aged 65.7 (10) years were analysed. Mean BP measurements for the physician using the mercury sphygmomanometer, the physician using the Omron, and the patient using the same device were: 136 (15.8)/80 (11), 137 (17.9)/80 (10), and 139* (17.6)/80 (10) mmHg, respectively. BP control was 48.1, 48.1, and 36.8*% (*P < 0.05), respectively. Intraclass correlation coefficients for systolic/diastolic pressures were: 0.77/0.65 (physician-sphygmomanometer Hg, physician-Omron; P < 0.001), 0.75/0.64 (physician-sphygmomanometer Hg, patient Omron, P < 0.001), and 0.83/0.83 (physician-Omron, patient-Omron; P < 0.001). In conclusion, the three types of measurement in the medical office were significantly concordant. Patient office self-measurement showed a tendency to increase systolic BP and worsen BP control. PMID- 12571617 TI - Self-blood-pressure monitoring--a questionnaire study: response, requirement, training, support-group popularity and recommendations. AB - The objective of this study was to survey hypertensive patients' response to, requirement for and training in self-blood pressure monitoring (SBPM). A total of 222 hypertensives were invited to complete a questionnaire even when not participating in the project. Questions supplied information on demographics, monitoring frequency, convenience of attending the surgery, monitor ownership and preference for and ease of self-monitoring. Comments supplied qualitative data. Training group questionnaires supplied similar data as well as SBPM data before and after training. Of 133 respondents, a higher educated, younger, wider age range wanted to participate (76; 57.2%) and tended to self-monitor. However, only an increase in further education (FE) was associated with an increased probability of participation and inclination to self-monitor in the multivariate analyses. A positive relationship exists between age and frequency in both groups. About a sixth of respondents own monitors and ownership is correlated to FE. Although most patients found it convenient to visit the surgery, the percentage finding it convenient was lower in patients attending training sessions than in the original survey, possibly indicating that independence is important for SBPM. Younger, higher-educated patients tended to self-monitor although FE was again the significant factor. Training increased preference for the idea of SBPM, prospective monitor ownership (64.8% of non-owners) and self monitoring intention (76.1%). Patients found recording card listed cardiovascular disease (CVD) risks valuable and 69.6% (32) wanted to establish a support group. Comments and interviews indicated haphazard knowledge, routines and uncertainty about SBPM. A standardised procedure including patient assessment, SBPM protocol and lifestyle education is needed for SBPM to be successful. PMID- 12571618 TI - HLA phenotypes as promoters of cardiovascular remodelling in subjects with arterial hypertension. AB - The objective of this study was to investigate the association between human leukocyte antigens (HLA) phenotypes and cardiovascular remodelling, as expressed by left ventricular mass (LVM) and carotid intima-media thickness (IMT), in hypertensives. We examined 153 subjects with arterial hypertension and 61 normotensive controls living in the greater Athens area. The population was classified into three groups and specifically group I (normotensives), group II with Grade 1 hypertension and group III with Grade 2 or 3 hypertension. HLA class I and class II antigens were studied by microlymphocytotoxic technique. Carotid IMT and LVM were determined by ultrasonography. The prevalence of HLA DQ7 in the hypertensive cohort was 27.4% that was significantly smaller than the 52.5% among the controls (P = 0.002). The HLA DR11 was found in 24.0% of the hypertensives and in 52.5% of the controls (P < 0.001). Group III hypertensives with HLA DR11 exhibited significantly higher LVM/h in comparison to the hypertensives without this HLA (199.0 +/- 28.8 vs 171.2+44.1g/m, P = 0.009). This association was not present in groups I and II. Similarly, group III hypertensives with HLA DQ7 were characterized by higher IMT in comparison to those without this HLA (0.94 +/- 0.19 vs 0.83 +/- 0.23 mm, P = 0.048). HLA DR17 was associated with higher IMT in both groups II and III (1.00 +/- 0.19 vs 0.82 +/- 0.19 mm, P = 0.046 and 1.01 +/- 0.23 vs 0.84 +/- 0.22 mm, P = 0.049, respectively) but not in group I. In conclusion, certain HLA phenotypes may be related to the levels of arterial blood pressure. Moreover, it seems that these HLA phenotypes may identify subjects with arterial hypertension that are more prone to develop cardiovascular hypertrophy. PMID- 12571619 TI - Linkage analysis of five candidate genes and essential hypertension in 106 Chinese nuclear families. AB - Five candidate genes including the lipoprotein lipase, leptin, leptin receptor, alpha-adducin and beta3 adrenergic receptor were selected to examine their possible contribution to essential hypertension (EH) in a Chinese population. On each side of the candidate gene loci, one to two highly polymorphic microsatellite markers were genotyped in 474 subjects recruited from 106 hypertension nuclear families in Shanghai. Both parametric and nonparametric linkage analyses were carried out using GENEHUNTER (version 2.0) after genotyping. Extended transmission disequilibrium testing (ETDT) was also conducted to detect preferential transmission of alleles to affected offspring. We failed to find the linkage between all these loci and EH by either parametric or nonparametric analysis, nor did we detect any significant transmission disequilibrium by ETDT. Our findings provide no support for a significant contribution of these five genes to the pathogenesis of EH among Shanghai people. PMID- 12571620 TI - Seizures as a presenting symptom of phaeochromocytoma in a young soldier. AB - New onset of seizures in young adults is frequently related to alcohol withdrawal, toxic exposure, central nervous system trauma and neoplasm. We describe a young soldier presenting to the emergency department with seizures and transient coma. On admission, he had hypertension, marked leukocytosis, hyper glycaemia, acidosis, elevated creatinine and elevated creatine phosphokinase of muscle origin. A thorough work-up revealed elevated urinary catecholamines, and a left adrenal mass was found on MRI and MIBG scan. The patient underwent laparoscopic adrenalectomy and completely recovered. This is the first description of seizures as a presenting symptom of phaeochromocytoma. PMID- 12571621 TI - Blood pressure rise in spontaneous intracerebral haemorrhage: epiphenomenon or precipitating factor? PMID- 12571626 TI - CAR is a cell-cell adhesion protein in human cancer cells and is expressionally modulated by dexamethasone, TNFalpha, and TGFbeta. AB - The coxsackie adenovirus receptor (CAR) has become of interest for gene therapy due to its crucial function in adenoviral cell entry. In clinical trials with adenoviral vectors, dexamethasone is applied to reduce side effects such as inflammatory reactions or emesis. By using a beta-galactosidase-expressing adenovirus (AdGal), we observed that dexamethasone treatment resulted in decreased adenoviral gene transfer into human cancer cells. Expression of CAR and integrin alpha5beta1 was transcriptionally downregulated by dexamethasone as shown for HeLa cervical cancer cells and U87MG glioblastoma cells. TNFalpha increased CAR expression in HeLa and ovarian cancer cells but decreased CAR expression in U87MG cells. In all tested cancer cell lines, TNFalpha induced a significant increase in the expression of adenovirus-binding integrins alpha5beta1, alphavbeta3 and alphavbeta5. Pretreatment with TNFalpha increased AdGal gene transfer into cancer cells and enhanced the cytotoxic effect of a p53 expressing adenovirus. In contrast, TGFbeta reduced CAR expression level and adenoviral gene transfer into OV-UL-2 ovarian cancer cells. Confocal immunofluorescence analysis revealed localization of CAR at cell-cell adhesions in several human cancer cell lines and disruption of cell-cell contacts increased adenoviral gene transfer into human cancer cells. In clinical cancer gene therapy, efficiency of adenoviral gene delivery could be altered by cell adhesion, TNFalpha, TGFbeta, and dexamethasone. PMID- 12571627 TI - Selective gene expression using a DF3/MUC1 promoter in a human esophageal adenocarcinoma model. AB - The efficacy of replication-deficient adenoviral vectors in gene therapy is confined to the number of tumor cells the vector infects. To focus and enhance the therapeutic efficacy, we employed a conditionally replication-competent adenoviral vector with a tissue-specific promoter, DF3/MUC1, in a human esophageal adenocarcinoma model. Our results demonstrate that Ad.DF3.E1A.CMV.TNF (Ad.DF3.TNF) specifically replicates in Bic-1 (DF3-producing cells) and mediates an enhanced biologic effect due to increased TNF-alpha in the same DF3-producing cells. We also show that the increased TNF-alpha interacts with ionizing radiation to produce greater tumor regression and a greater delay in tumor regrowth in Bic-1 (DF3-producing cells) compared to Seg-1 (DF3 non-producers). Tumor cell targeting using conditionally replication-competent adenoviral vectors with tumor-specific promoters to drive viral replication and deliver TNF-alpha provides a novel approach to enhancing tumor radiosensitivity. PMID- 12571628 TI - Airway-directed gene transfer of interleukin-10 using recombinant Sendai virus effectively prevents post-transplant fibrous airway obliteration in mice. AB - Bronchiolitis obliterans (BO) after lung transplantation prevents a satisfactory prognosis, and recent studies suggested that interleukin-10 (IL-10) gene transfer to distant organs could inhibit BO in rodent models. Although delivery of the therapeutic gene to a local airway would be favored to minimize systemic effects, current limitations include lower gene transfer efficiency to airway epithelium. As recombinant Sendai virus (SeV) can produce dramatically efficient gene transfer to airway epithelium, we determined if SeV-mediated IL-10 gene transfer to the local airway would inhibit bronchial fibrous obliteration in murine tracheal allografts. Administration of cyclosporine A (CsA) significantly promoted not only recovery of the injured airway epithelium but also SeV-mediated IL-10 expression (CsA- versus CsA+ =228+/-78 versus 3627+/-1372 pg/graft with 5 x 10(7) pfu), thereby suggesting the requirement of epithelia for efficient gene transfer. Even at the highest expression, no significant leakage of IL-10 was evident in the systemic circulation, and the induction of interferon-gamma was completely diminished on day 7 by IL-10 gene transfer. As a result, luminal loss was significantly prevented in allografts treated with SeV-IL-10 (luminal opening, all control groups: 0% respectively, and SeV-IL-10 5 x 10(7) pfu: 25.7+/ 10.5%), an effect that was enhanced by short-term CsA treatment (SeV-IL-10 5 x 10(7) pfu with CsA: 63.7+/-12.7%). We propose that SeV is a useful vector that can target airway epithelium to prevent BO avoiding putative systemic effect. PMID- 12571629 TI - Lentivirus-mediated expression of angiostatin efficiently inhibits neovascularization in a murine proliferative retinopathy model. AB - Ischemic retinal diseases, such as diabetic retinopathy, retinopathy of prematurity, and age-related macular degeneration, are a major cause of blindness worldwide. Angiostatin is an internal peptide fragment of plasminogen that inhibits endothelial proliferation in vitro and tumor growth in vivo. We now demonstrate that HIV vector encoding angiostatin (HIV-angiostatin) can inhibit retinal neovascularization in a mouse model of proliferative retinopathy. Intravitreal injections of HIV-angiostatin led to stable expression of the angiostatin gene in retinal tissue. Retinal neovascularization was histologically quantitated by a masked protocol. Retinal neovascularization in the eye injected with HIV-angiostatin was reduced in 90% (9/10; P=0.025) of animals, compared with the eye injected with phosphate-buffered saline. Reduction of histologically evident neovascular nuclei per 6-microm section averaged 68%, with maximal inhibitory effects of 87%. Neovascularization was not reduced in the eyes injected with HIV vector encoding enhanced green fluorescent protein. This is the first report that HIV-angiostatin can reduce neovascular cell nuclei in a murine proliferative retinopathy model. These data suggest that the anti-angiogenic activity of angiostatin has therapeutic potential for the treatment of retinal neovascularization. PMID- 12571630 TI - Gamma-irradiation enhances transgene expression in leukemic cells. AB - The majority of immunotherapy-based gene therapy protocols consist of ex vivo gene transfer in tumor cells. To prevent further in vivo growth, modified cells must be irradiated before reinjection into patients. The present study examines the effects of gamma-irradiation on transgene expression in transduced leukemic cells. Human and murine leukemic cells were transfected with retroviral vectors or plasmids carrying beta-galactosidase, GM-CSF or CD80 genes. Fresh leukemic cells from patients with acute myeloid leukemia (AML) were transfected with AdZ.F(pK7) adenoviral vector. gamma-irradiation at various lethal doses enhanced transgene expression in leukemic cell lines and fresh AML cells when the gene of interest was under CMV promoter but not when SV40 promoter was used. Oxidative stress also enhanced transgene expression and both irradiation and oxidative stress effects were inhibited by addition of N-acetyl-L-cysteine, a thiol anti oxidant, indicating the involvement of reactive oxygen species. Transgene expression was also enhanced in vivo 48 and 120 h after subcutaneous injection of irradiated leukemic cells in syngeneic mice. These results show that a cell vaccine protocol using ex vivo gene transfer of transduced cells might be feasible in acute leukemia even if leukemic cells must be irradiated at lethal doses prior to reinjection to patients. PMID- 12571631 TI - Cartilage degradation and invasion by rheumatoid synovial fibroblasts is inhibited by gene transfer of TIMP-1 and TIMP-3. AB - Matrix metalloproteinases (MMPs) are believed to be pivotal enzymes in the invasion of articular cartilage by synovial tissue in rheumatoid arthritis (RA). Here, we investigated the effects of gene transfer of tissue inhibitors of metalloproteinases (TIMPs) on the invasiveness of RA synovial fibroblasts (RASF) in vitro and in vivo. Adenoviral vectors (Ad) were used for gene transfer. The effects of AdTIMP-1 and AdTIMP-3 gene transfer on matrix invasion were investigated in vitro in a transwell system. Cartilage invasion in vivo was studied in the SCID mouse co-implantation model for 60 days. In addition, the effects of AdTIMP-1 and AdTIMP-3 on cell proliferation were investigated. A significant reduction in invasiveness was demonstrated in vitro as well as in vivo in both the AdTIMP-1- and AdTIMP-3-transduced RASF compared with untransduced SF or SF that were transduced with control vectors. in vitro, the number of invading cells was reduced to 25% (P<0.001) in the AdTIMP-1-transduced cells and to 13% (P<0.0001) in the AdTIMP-3-transduced cells (% of untransduced cells). Cell proliferation was significantly inhibited by AdTIMP-3 and, less, by AdTIMP-1. In conclusion, overexpression of TIMP-1 and TIMP-3 by Ad gene transfer results in a marked reduction of the invasiveness of RASF in vitro and in the SCID mouse model. Apart from the inhibition of MMPs, a reduction in proliferation rate may contribute to this effect. These results suggest that overexpression of TIMPs, particularly TIMP-3 at the invasive front of pannus tissue, may provide a novel therapeutic strategy for inhibiting joint destruction in RA. PMID- 12571632 TI - Priming of T cells with Ad-transduced DC followed by expansion with peptide pulsed DC significantly enhances the induction of tumor-specific CD8+ T cells: implications for an efficient vaccination strategy. AB - In recent years, vaccination strategies using antigen-presenting cells (APC) have been under investigation. Antigen delivery using genetic immunization through ex vivo transduction of dendritic cells (DC) is supposed to enhance the induction of antitumor responses in humans by activating a broad range of peptide-specific CD8+ T cells. In this study, we compared the potential of adenoviral (Ad) transduced versus peptide-pulsed DC to induce melanoma-antigen (Ag)-specific T cell responses in vitro. Whereas gp100-peptide-pulsed DC induced long-lasting specific CD8+ T-cell responses against single peptides, Ad-transduced DC induced broad and strong, specific immunity against various peptides of the gp100-Ag. Surprisingly, several restimulations led to decreasing gp100-specific and in parallel to increasing anti-adenoviral T-cell responses. Nevertheless, those anti adenoviral T-cell responses provided an "adjuvant" effect by inducing an early release of high amounts of IL-2/IFN-gamma, therewith enhancing CTL induction in the initiation phase. Based on these data, we suggest a prime/boost vaccination strategy in melanoma patients--combining the use of Ad-DC and peptide-pulsed DC- to obtain efficient and long-term antitumor T-cell responses. PMID- 12571633 TI - Enabling topical immunization via microporation: a novel method for pain-free and needle-free delivery of adenovirus-based vaccines. AB - The skin represents an excellent site for vaccine inoculation due to its natural role as a first line of contact with foreign pathogens and the high local frequency of antigen presenting cells. To facilitate skin-directed immunization, a new technique has been developed (termed microporation) whereby a vaporization process is used to remove tiny areas of the stratum corneum creating microscopic pores that allow access to the underlying viable epidermis. Reporter gene expression was 100-fold increased following application of an adenovirus vector to microporated skin when compared to intact skin. Furthermore, 10-100-fold greater cellular and humoral immune responses were observed following topical administration of an adenovirus vaccine to microporated skin versus intact skin. Hairless mice responded to the microporated adenovirus vaccine equivalently to mice with normal hair follicle distribution demonstrating the activity of the microporated vaccine was not related to follicle count. In a tumor challenge model using a surrogate antigen, microporation increased vaccine efficacy by approximately 100-fold compared to intact skin. Finally, microporation enabled delivery of an adenovirus vaccine carrying a relevant melanoma antigen resulting in the development of auto-immune vitiligo and tumor protection. Thus, the microporation technology has proven to be a reliable and easy method to enable skin-directed vaccination. PMID- 12571634 TI - Recharging cationic DNA complexes with highly charged polyanions for in vitro and in vivo gene delivery. AB - The intravenous delivery of plasmid DNA complexed with either cationic lipids (CL) or polyethyleneimine (PEI) enables high levels of foreign gene expression in lung. However, these cationic DNA complexes cause substantial toxicity. The present study found that the inclusion of polyacrylic acid (pAA) with DNA/polycation and DNA/CL complexes prevented the serum inhibition of the transfection complexes in cultured cells. The mechanism mediating this increase seems to involve both particle size enlargement due to flocculation and electrostatic shielding from opsonizing serum proteins. The use of pAA also increased the levels of lung expression in mice in vivo substantially above the levels achieved with just binary complexes of DNA and linear PEI (lPEI) or CL and reduced their toxicity. Also, the use of a "chaser" injection of pAA 30 min after injection of the ternary DNA/lPEI/pAA complexes further aided this effort to reduce toxicity while not affecting foreign gene expression. By optimizing the amount of pAA, lPEI, and DNA within the ternary complexes and using the "chaser" injection, substantial levels of lung expression were obtained while avoiding adverse effects in lung or liver. These developments will aid the use of cationic DNA complexes in animals and for eventual human gene therapy. PMID- 12571635 TI - Recombinant Sendai virus provides a highly efficient gene transfer into human cord blood-derived hematopoietic stem cells. AB - Hematopoietic stem cells (HSCs) are a promising target for gene therapy, however, the low efficiencies of gene transfer using currently available vectors face practical limitations. We have recently developed a novel and efficient gene transfer agent, namely recombinant Sendai virus (SeV), and we have here characterized SeV-mediated gene transfer to human cord blood (CB) HSCs and primitive progenitor cells (PPC) using the jelly fish green fluorescent protein (GFP) gene. Even at a relatively low titer (10 multiplicity of infections), SeV achieved highly efficient GFP expression in CB CD34(+) cells (85.5+/-5.8%), as well as more immature CB progenitor cells, CD34(+)AC133(+) (88.2+/-3.7%) and CD34(+)CD38(-) (84.6+/-5.7%) cells, without cytokines prestimulation, that was a clear contrast to the features of gene transfer using retroviruses. SeV-mediated gene transfer was not seriously affected by the cell cycle status. In vitro cell differentiation studies revealed that gene transfer occurred in progenitor cells of all lineages (GM-CFU, 73.0+/-11.1%; BFU-E, 24.7+/-4.0%; Mix-CFU, 59+/-4.0%; and total, 50.0+/-7.0%). These findings show that SeV could prove to be a promising vector for efficient gene transfer to CB HSCs, while preserving their ability to reconstitute the entire hematopoietic series. PMID- 12571636 TI - Protein delivery by subviral particles of human cytomegalovirus. AB - Direct protein delivery is an emerging technology in vaccine development and gene therapy. We could previously show that subviral dense bodies (DB) of human cytomegalovirus (HCMV), a beta-herpesvirus, transport viral proteins into target cells by membrane fusion. Thus these non-infectious particles provide a candidate delivery system for the prophylactic and therapeutic application of proteins. Here we provide proof of principle that DB can be modified genetically. A 55 kDa fusion protein consisting of the green fluorescent protein and the neomycin phosphotransferase could be packed in and delivered into cells by recombinant DB in a functional fashion. Furthermore, transfer of protein into fibroblasts and dendritic cells by DB was efficient, leading to exogenous loading of the MHC class I antigen presentation pathway. Thus, DB may be a promising basis for the development of novel vaccine strategies and therapeutics based on recombinant polypeptides. PMID- 12571637 TI - Gene therapy progress and prospects: alpha-1 antitrypsin. AB - Over the 2 years covered here, there has been one clinical study in which a normal alpha-1 antitrypsin (AAT) gene was delivered to the nasal epithelium of AAT-deficient subjects using plasmid-liposome complexes; a second study using an adeno-associated vector should begin soon. Although progress in clinical studies has been slow, advances in both viral and nonviral vector designs show considerable promise. Strategies that combine liposome technology with imaginative vector design may permit long-term expression of a normal transgene that is sufficient to achieve therapeutic serum AAT concentrations. While reproducing the normal physiology by targeting normal AAT gene expression to the liver is logical, local expression in lung cells may be less demanding of the technology and offers therapeutic benefits that are produced neither by AAT protein therapy nor by AAT gene therapy targeted to the liver. Developing technologies may permit direct correction of the mutant AAT gene using innovative approaches to in vivo gene repair. PMID- 12571638 TI - Recombinant adenovirus-mediated cytotoxic gene therapy of lymphoproliferative disorders: is CAR important for the vector to ride? AB - The literature has seen an incredible booming of publications related to the use of recombinant adenoviruses as therapeutic tools for lymphoproliferative disorders over the last decade. Several approaches of adenovirus-mediated gene expression have been used to transfect cell lines that are derived from lymphoid tumors and would have otherwise been refractory to other transfection methods. The identification of high-affinity receptor for human adenoviruses serotype 2 and 5, the coxsackie-adenovirus receptor (CAR), has raised the question about its relevance for the efficacy of recombinant adenovirus-mediated gene therapy. We review published studies that have analyzed the use of recombinant adenovirus vectors expressing cytotoxic genes for gene therapy in lymphomas, chronic lymphocytic leukemia and multiple myeloma. For simplicity, we group all these diseases under the term lymphoproliferative disorders. We analyze the use of recombinant adenovirus-mediated cytotoxicity by assessing the importance of the biochemical and intrinsic signaling pathways interacting with the products of the exogenous viral-mediated expression. Ultimately, we discuss studies that have been finalized to by-pass the limitations of the biodistribution of CAR by modifying or targeting adenovirus to other membrane proteins in cells derived from lymphoproliferative disorders. PMID- 12571639 TI - Non-invasive gamma camera imaging of gene transfer using an adenoviral vector encoding an epitope-tagged receptor as a reporter. AB - A model epitope-tagged receptor was constructed by fusing the hemagglutinin (HA) sequence on the extracellular N-terminus of the human somatostatin receptor subtype 2 (hSSTr2) gene. This construct was placed in an adenoviral (Ad-HAhSSTr2) vector. This study evaluated Ad-HAhSSTr2 in vitro and in vivo using FACS, fluorescent microscopy, radioactive binding assays, and gamma camera imaging techniques. Infection of A-427 non-small cell lung cancer cells with Ad-HAhSSTr2 or Ad-hSSTr2 resulted in similar expression of hSSTr2 by FACS analysis and binding assays using a (99m)Tc-labeled somatostatin analogue ((99m)Tc-P2045). HAhSSTr2 expression in A-427 cells was specific for infection with Ad-HAhSSTr2. FITC-labeled anti-HA antibody (FITC-HA) confirmed surface expression in live A 427 cells and the absence of internalization. Gamma camera imaging and gamma counter analysis of normal mice showed significantly greater (P<0.05) liver uptake of (99m)Tc-labeled anti-HA antibody ((99m)Tc-anti-HA) in mice injected i.v. 48 h earlier with Ad-HAhSSTr2 (53.6+/-6.9% ID/g) as compared to mice similarly injected with Ad-hSSTr2 (9.0+/-1.3% ID/g). In a mouse tumor model, imaging detected increased tumor localization of (99m)Tc-anti-HA due to direct intratumor injection Ad-HAhSSTr2. Gamma counter analysis confirmed significantly greater (P<0.05) uptake of (99m)Tc-anti-HA in tumors injected with Ad-HAhSSTr2 (12.5+/-4.1% ID/g) as compared to Ad-hSSTr2-infected tumors (5.1+/-1.5% ID/g). These studies demonstrate the feasibility of using an epitope-tagged reporter receptor for non-invasively imaging gene transfer. PMID- 12571640 TI - Adeno-associated virus-mediated delivery of BCL-w gene improves outcome after transient focal cerebral ischemia. AB - A recombinant adeno-associated virus (rAAV) vector was used to overexpress the anti-apoptotic Bcl-2-family protein, BCL-w, in rat brain. Three weeks after injecting the vector into cerebral cortex and striatum on one side, temporary focal ischemia was induced by occlusion of the ipsilateral middle cerebral artery for 90 min, followed by reperfusion for 24 h. BCL-w expression was increased in cerebral cortex and striatum--and in neurons, astroglia and endothelial cells--in the brains of rats that received the rAAV-BCL-w vector, compared to rats given phosphate-buffered saline or a control vector containing the gene for green fluorescent protein. Recipients of the rAAV-BCL-w vector also showed a 30% reduction in infarct size and a 33-40% improvement in neurological function, compared to the control groups. These results provide evidence for a role of BCL w in regulating histological and functional outcome after focal cerebral ischemia. PMID- 12571641 TI - Endostatin gene therapy on murine lung metastases model utilizing cationic vector mediated intravenous gene delivery. AB - Tumors require ongoing angiogenesis to support their growth. Inhibition of angiogenesis by production of antiangiogenic factors should be a viable approach for cancer gene therapy. In this study, we investigated whether intravenous administration of endostatin gene complexed with a cationic vector (GL67/DOPE or PEI22K) could inhibit the development of lung tumors in mice injected i.v. with NFSa Y83 fibrosarcoma cells (5 x 10(5)) which frequently form lung metastasis. mRNA and protein of the transfected gene were produced in the lung and other organs of the transfected mice as assessed by immunohistochemistry, Western blotting and reverse transcription-polymerase chain reaction. Single intravenous injection of the endostatin gene (60 microg) complexed with either GL67/DOPE or PEI22K on day 3 or day 7 after fibrosarcoma cell inoculation significantly inhibited tumor formation in the lung as evidenced by the reduced number of lung tumors and lung weight, and prolonged survival of the endostatin gene-transfected mice compared with control mice. These findings suggested that the endostatin gene therapy, using cationic vector-mediated intravenous gene transfer, might be a feasible strategy for organ-targeted prevention and regulation of possible disseminated cancers. PMID- 12571642 TI - Non-viral gene delivery in skeletal muscle: a protein factory. AB - Ever since the publication of the first reports in 1990 using skeletal muscle as a direct target for expressing foreign transgenes, an avalanche of papers has identified a variety of proteins that can be synthesized and correctly processed by skeletal muscle. The impetus to the development of such applications is not only amelioration of muscle diseases, but also a range of therapeutic applications, from immunization to delivery of therapeutic proteins, such as clotting factors and hormones. Although the most efficient way of introducing transgenes into muscle fibres has been by a variety of recombinant viral vectors, there are potential benefits in the use of non-viral vectors. In this review we assess the recent advances in construction and delivery of naked plasmid DNA to skeletal muscle and highlight the options available for further improvements to raise efficiency to therapeutic levels. PMID- 12571643 TI - Dendritic cells genetically engineered to express IL-4 exhibit enhanced IL-12p70 production in response to CD40 ligation and accelerate organ allograft rejection. AB - C57BL/10 (B10; H2(b)) bone marrow-derived myeloid dendritic cells (DC) propagated in GM-CSF + IL-4 were transduced with r adenoviral (Ad) vectors encoding either control neomycin-resistance gene (Ad-Neo) or murine IL-4 (Ad-IL-4) on day 5 of culture following CD11c immunomagnetic bead purification. Both Ad-Neo- and Ad-IL 4-transduced DC displayed upregulated surface MHC class II and costimulatory molecules (CD40, CD80, CD86). Ad-IL-4 DC secreted higher levels of bioactive IL 12p70 after CD40 ligation or LPS stimulation than either Ad-Neo or unmodified DC. Only Ad-IL-4 DC produced IL-12p70 in primary MLR, in which they induced augmented proliferative responses of naive allogeneic C3H/HeJ (C3H; H2(k)) T-cells. Compared with Ad-Neo DC, Ad-IL-4 DC were also more effective in priming naive allogeneic recipients to exhibit specifically enhanced anti-donor T-cell proliferative and CTL responses. T-cells primed in vivo 7 days previously with Ad IL-4 DC displayed enhanced secretion of Th2 (IL-4, IL-10) but also higher Th1 cytokine (IFNgamma) production following ex vivo challenge with donor alloAg. Moreover, pretreatment of vascularized heart graft recipients with i.v. Ad-IL-4 DC, 1 week before transplant, significantly accelerated rejection and antagonized the therapeutic effect of anti-CD40L (CD154) mAb. These data contrast markedly with recently reported inhibitory effects of autologous Ad-IL-4 DC on autoimmune inflammatory disease. PMID- 12571644 TI - Genetic manipulations of adenovirus type 5 fiber resulting in liver tropism attenuation. AB - The development of genetically modified adenoviral vectors capable of specifically transducing a given cell population requires the addition and functional presentation of particular tropism determinants within the virus capsid, together with the abrogation of the molecular determinants that dictate their natural tropism in vivo. The human adenovirus serotype 5 (Ad5) first attaches to the cell surface following high-affinity binding of the C-terminal knob of the fiber capsid protein to the coxsackie and adenovirus receptor (CAR). Here we have assessed whether genetic shortening of the fiber shaft (virus BS1), or replacing the Ad5 fiber shaft and knob with their Ad3 counterparts (virus DB6), could cripple this interaction in vitro and in vivo. A 10-fold decrease in the binding of the modified capsids to soluble CAR was evidenced, which correlated with a similar reduction of their ability to transduce CAR-positive cells in vitro. The ability of BS1 to interact with cellular integrins was also impaired, suggesting that the penton base and the short-shafted fiber when embedded in the capsid preclude each other from efficiently interacting with their cognate cell surface receptors (CAR and integrins respectively). BS1 and DB6 intravenous injections in mice further supported a profound impairment of the ability of the capsid-modified viruses to transduce the liver as demonstrated by a 10-fold reduction of intracellular viral DNA and transgene expression. Interestingly enough, the host humoral response was also specifically weakened in BS1- and DB6-inoculated animals. Taken together, these observations indicate that (i) fiber shortening and (ii) pseudo-typing of Ad5-based vectors with the shaft and knob from non-CAR-binding serotypes constitute two promising strategies to successfully attenuate their native tropism in vitro and most importantly in vivo. PMID- 12571645 TI - Cell phenotype specific kinetics of expression of intratracheally injected manganese superoxide dismutase-plasmid/liposomes (MnSOD-PL) during lung radioprotective gene therapy. AB - Intratracheal (IT) injection of manganese superoxide dismutase-plasmid/liposome (MnSOD-PL) complexes prior to whole lung irradiation of C57BL/6J mice provides significant protection from acute and chronic irradiation damage. We determined the duration of increased MnSOD biochemical activity and differential expression of a hemagglutinin (HA) epitope-tagged MnSOD transgene. HA-MnSOD-PL was IT injected at doses of 0-1000 microg, and mice were killed 1,2,3 or 4 days later. Other groups of mice were irradiated to 20 Gy to the pulmonary cavity 24 h after injection and killed at the same time points as non-irradiated mice. Both non irradiated and irradiated groups of mice showed increased MnSOD biochemical activity with plasmid dose that plateaued at 100 microg of MnSOD plasmid DNA. In control mice, MnSOD biochemical activity decreased at 2, 3 or 4 days after injection. In irradiated mice, MnSOD biochemical activity decreased at day 2 but increased on days 3 and 4. HA-MnSOD expression decreased in broncheoalveolar macrophages and alveolar type-II cells 3 days after injection in non-irradiated and irradiated mice, but remained elevated in endothelial and epithelial cells past 4 days. The data provide a rationale for every second-day administration of intrapulmonary MnSOD-PL in clinical trials of radioprotective gene therapy. This should be sufficient to provide radioprotection during radiation treatments. PMID- 12571646 TI - Adenovirus-mediated gene therapy for bladder cancer: efficient gene delivery to normal and malignant human urothelial cells in vitro and ex vivo. AB - Existing local therapies for superficial transitional cell carcinoma (TCC) of the bladder have limited success in preventing progression to life-threatening, muscle-invasive disease, and novel therapies are needed. Recent studies have raised doubts concerning the feasibility of adenovirus-mediated gene therapy for bladder cancer. We have therefore investigated adenoviral transduction of normal and malignant human urothelial cells, both as primary cultures and in intact epithelium. All 15 primary normal human urothelial cell lines tested were transduced in vitro by Adv-cmv-beta-gal at high efficiency, and better than most human TCC cell lines. Eight primary human TCC explants were also successfully transduced. In contrast, in intact normal urothelium, transduction efficiency was lower, and occurred only in superficial epithelial layers. Expression of the hCAR adenovirus receptor, however, occurred throughout the full thickness of urothelium. Transduction of human TCC biopsy specimens was at least as efficient as intact normal urothelium.We demonstrate for the first time that adenoviral transduction of both normal and malignant human urothelial cells is feasible. A physical barrier, rather than hCAR status, may be the main determinant of transduction of intact epithelium. Clinical trials of adenovirus-mediated gene therapy for superficial bladder cancer are warranted. PMID- 12571647 TI - Poly(cationic lipid)-mediated in vivo gene delivery to mouse liver. AB - We have previously demonstrated that liposomes generated from poly(cationic lipid) (PCL) and cholesterol (Chol) have low cytotoxicity, are serum resistant, and display a transfection efficiency in vitro similar to commercially available cationic liposomes. Our in vivo experiments demonstrated that PCL-Chol liposomes bound much less avidly to serum proteins than did liposomes composed of 1,2 bis(dioleoyloxy)-3-(trimethylamonio)propane (DOTAP)-Chol or DOTAP-L-alpha dioleoyl phosphatidylethanolamine (DOPE). Injection of the lipoplexes (PCL Chol+DNA) through the portal vein after partial hepatectomy (PH) led to much higher reporter gene expression (luciferase) in the liver than did naked DNA injection. Marked green fluorescent protein expression was visualized in almost all hepatocytes in the liver of mice receiving lipoplex injection, even in the absence of PH. Subcutaneous injection of thyroid hormone triiodothyromine (T(3)) significantly promoted hepatocyte regeneration and markedly enhanced PCL-Chol mediated gene transfer in mouse liver when the lipoplex was administrated through either portal or tail vein. With T(3) pretreatment, PCL-Chol exerted a better gene transfer efficacy in mouse liver than DOTAP-Chol or DOTAP-DOPE. Two injections of lipoplexes through an indwelling catheter in the portal vein extended the transgene expression at a high level when T(3) injection was repeated. In conclusion, our findings demonstrate that the polymerized cationic liposomes are very stable in the blood and are effective agents for in vivo gene delivery, and that thyroid hormone administration offers a non-invasive approach to enhance liposome-mediated liver gene delivery. PMID- 12571648 TI - Dexamethasone inhibits the anti-tumor effect of interleukin 4 on rat experimental gliomas. AB - Retroviral-mediated gene transfer of the IL-4 gene into experimental gliomas can cause tumor rejection, supporting the clinical use of this form of gene therapy for glioblastomas (GBM). In a clinical setting, the administration of dexamethasone (dex) is a standard procedure for GBM patients. This led us to examine the effects of dex on IL-4 gene therapy. We injected intracranially Fischer 344 rats with phosphate-buffered saline, 9L gliosarcoma cells mixed with E86.L4SN(200) cells (retroviral producer cells, RPC, transducing IL-4 cDNA) and 9L cells mixed with PA317.STK.SBA cells (control RPC expressing the HSV-tk gene). The rats from each group were treated with 0, 50, 100 or 250 microg dex/kg/day released by osmotic pumps implanted subcutaneously. While 80-100% of rats receiving 9L cells mixed with IL-4 RPC and not treated by dex survived for at least 2 months following tumor injection, only 50% and 17% of rats receiving 50 or 100 microg/kg/day of dex, respectively, reached this time point. These results indicate that dex significantly diminished the anti-tumor effect of IL-4. Thus, in a clinical setting, IL-4 gene transfer should be performed when low levels of dex are administered or in the absence of dex. PMID- 12571649 TI - Lack of benefit of dietary advice to men with angina: results of a controlled trial. AB - OBJECTIVE: To see whether mortality among men with angina can be reduced by dietary advice. DESIGN: A randomized controlled factorial trial. SETTING: Male patients of general practitioners in south Wales. SUBJECTS: A total of 3114 men under 70 y of age with angina. INTERVENTIONS: Subjects were randomly allocated to four groups: (1) advised to eat two portions of oily fish each week, or to take three fish oil capsules daily; (2) advised to eat more fruit, vegetables and oats; (3) given both the above types of advice; and (4) given no specific dietary advice. Mortality was ascertained after 3-9 y. RESULTS: Compliance was better with the fish advice than with the fruit advice. All-cause mortality was not reduced by either form of advice, and no other effects were attributable to fruit advice. Risk of cardiac death was higher among subjects advised to take oily fish than among those not so advised; the adjusted hazard ratio was 1.26 (95% confidence interval 1.00, 1.58; P=0.047), and even greater for sudden cardiac death (1.54; 95% CI 1.06, 2.23; P=0.025). The excess risk was largely located among the subgroup given fish oil capsules. There was no evidence that it was due to interactions with medication. CONCLUSIONS: Advice to eat more fruit was poorly complied with and had no detectable effect on mortality. Men advised to eat oily fish, and particularly those supplied with fish oil capsules, had a higher risk of cardiac death. This result is unexplained; it may arise from risk compensation or some other effect on patients' or doctors' behaviour. PMID- 12571650 TI - A simple estimate of mortality attributable to excess weight in the European Union. AB - OBJECTIVE: To estimate the current burden of mortality attributable to excess weight in the European Union (EU). METHODS: Prevalence of overweight (body mass index, BMI 25-29.9 kg/m(2)) and obesity (BMI >or=30 kg/m(2)) were based on self reported data from a survey with samples representative of the 15 EU Member States in 1997. Primary source of relative risk (RR) of death by BMI was the first American Cancer Prevention Study (CPS I). Additional calculations were performed to account for effect of smoking (using CPS I data for non- or never smokers), for pre-existing illness (using the second CPS, CPS II, data for healthy never-smokers) and using RRs derived from European rather than US data (using data from a meta-analysis of prospective studies). Mortality attributable to excess weight was calculated by combining the prevalences of overweight and obesity, the RRs, and the number of deaths in the EU countries. RESULTS: Annual deaths attributable to overweight and obesity totalled approximately 279 000 when RRs for all subjects were used. When RRs for nonsmokers only were applied to the entire population, about 304 000 deaths were attributable to excess weight. In analyses using RRs which controlled for both smoking and history of disease, the number of deaths attributable to excess weight was estimated at about 337 000 based on European data and at about 401 000 based on US data. In the EU, therefore, a minimum of 279 000 deaths were attributable to excess weight (7.7% of all deaths, varying from 5.8% for France through 8.7% for the UK). More attributable deaths occurred among the obese (175 000) than among the overweight (104 000). Around 70% were cardiovascular disease deaths (195 000) and 20% cancer deaths (53 000). CONCLUSION: Mortality attributable to excess weight is a major public health problem in the EU. At least one in 13 annual deaths in the EU are likely to be related to excess weight. PMID- 12571651 TI - Calculating energy requirements for men with HIV/AIDS in the era of highly active antiretroviral therapy. AB - OBJECTIVES: 1. To determine if resting energy expenditure (REE) adjusted for body composition is elevated in HIV-positive males when compared with healthy controls in the era of highly active antiretroviral therapy. 2. To examine the accuracy of prediction equations for estimating REE in people with HIV. 3. To determine if REE adjusting for body composition is significantly different between those HIV positive subjects reporting lipodystrophy (LD) or weight loss (>or=5%) and those who are weight stable when compared to controls. DESIGN: Cross-sectional study. SETTING: Tertiary referral hospital HIV unit and an outpatient clinic specializing in HIV care. SUBJECTS: HIV-positive males (n=70) and healthy male controls (n=16). METHODS: REE was measured using indirect calorimetry. Body composition was assessed using bioelectrical impedance analysis. RESULTS: 1. REE when adjusted for fat-free mass and fat mass using the general linear model (analysis of covariance) was greater in HIV-positive subjects than controls (7258+/-810 kJ, n=70 vs 6615+/-695 kJ, n=16, P<0.05). 2. The Harris and Benedict, Schofield, Cunningham and the two equations previously published by Melchior and colleagues in HIV-positive subjects all gave an estimate of REE significantly different from the measured REE in the HIV-positive subjects, therefore a new prediction equation was developed. The inability of the published equations to predict REE in the different HIV-positive subgroups reflected the heterogeneity in body composition. 3. REE adjusted for fat-free and fat mass was significantly greater in the both the HIV patients who were weight stable and those with lipodystrophy compared with the healthy controls. CONCLUSION: REE is significantly higher in HIV-positive males when compared with healthy controls. Body composition abnormalities common in HIV render the use of standard prediction equations for estimating REE invalid. When measuring REE in HIV positive males adjustment steps should include fat-free and fat mass. PMID- 12571652 TI - Can perinatal supplementation of long-chain polyunsaturated fatty acids prevent diabetes mellitus? AB - It is suggested that the negative correlation between breast-feeding and insulin resistance and diabetes mellitus can be related to the presence of significant amounts of long-chain polyunsaturated fatty acids in the human breast milk. Based on this, it is proposed that provision of adequate amounts of long chain polyunsaturated fatty acids during the critical periods of brain growth and development can prevent or postpone the development diabetes mellitus. PMID- 12571653 TI - Low prevalence of the metabolic syndrome in wine drinkers--is it the alcohol beverage or the lifestyle? AB - OBJECTIVE: To study how the intake of alcohol and the choice of wine, beer, and spirits is related to lifestyle factors and the metabolic syndrome in 60-y-old men and women. DESIGN: Cross-sectional population based study. SETTING: Stockholm County, Sweden. SUBJECTS: Sixty-year-old men and women (n=4232). RESULTS: Moderate intake of wine (10-30 g/day) was associated with a lifestyle characterized by being married, having a university education, being employed, being Swedish-born, having a good quality of life according to economy, leisure time and health, compared with a group with low alcohol intake. The opposite characteristics were seen among the non-drinkers. Drinkers of spirits were more often smokers and also reported higher intake of sausage and fried potatoes compared with a group with low alcohol intake. In women, the metabolic syndrome was significantly more common in non-drinkers (20%), P<0.05, and less common among wine drinkers (8%), P<0.01, compared with a group with low alcohol intake. After adjustments, a significant lower odds ratio for the metabolic syndrome were seen in wine drinkers in women (OR=0.60, P<0.05). CONCLUSIONS: Compared with low alcohol drinkers, moderate wine drinkers exhibited a more favorable pattern according to both lifestyle factors and metabolic parameters. The close link between alcohol drinking behaviour and lifestyle habits illustrate the complex relationship between alcohol and health. PMID- 12571654 TI - Bioavailability in humans of the flavanones hesperidin and narirutin after the ingestion of two doses of orange juice. AB - OBJECTIVE: Flavanones are polyphenols specific of citrus fruits, where they are present in high amounts. Although citrus fruits and juices are widely consumed in the world, little information has been published on flavanone bioavailability in humans. The aim of the present study was to determine the nature of the circulating metabolites, the plasma kinetics and the urinary excretion patterns of the flavanones, hesperidin and narirutin. DESIGN: After an overnight fast, five healthy volunteers ingested 0.5 or 1 l of a commercial orange juice providing 444 mg/l hesperidin and 96.4 mg/l narirutin, together with a polyphenol free breakfast. Blood was sampled at 10 different timepoints over a 24 h period. Urine was collected for 48 h, in five fractions. RESULTS: Flavanones metabolites appeared in plasma 3 h after the juice ingestion, reached a peak between 5 and 7 h, then returned to baseline at 24 h. The peak plasma concentration of hesperetin was 0.46+/-0.07 micro mol/l and 1.28+/-0.13 micro mol/l after the 0.5 and 1 l intake, respectively. It was lower for naringenin: 0.20+/-0.04 micro mol/l after the 1 l dose. The circulating forms of hesperetin were glucuronides (87%) and sulphoglucuronides (13%). For both flavanones, the urinary excretion was nearly complete 24 h after the orange juice ingestion. The relative urinary excretion was similar for hesperetin and naringenin and did not depend on the dose: values ranged from 4.1+/-1.2 to 7.9+/-1.7% of the intake. CONCLUSIONS: In case of a moderate or high consumption of orange juice, flavanones may represent an important part of the pool of total polyphenols present in plasma. PMID- 12571655 TI - Variable effects of American ginseng: a batch of American ginseng (Panax quinquefolius L.) with a depressed ginsenoside profile does not affect postprandial glycemia. AB - BACKGROUND: We have repeatedly reported that American ginseng (AG) with a specific ginsenoside profile significantly decreases postprandial glycemia. Whether this effect is reproducible using AG with a different profile is unknown. We therefore investigated the effect of a different batch of AG on glycemia following a 75 g oral glucose tolerance test (OGTT). METHODS: Using a randomized, single blind design, 12 normal subjects (six males and six females, aged 31+/-3 y, body mass index (BMI) 28+/-2 kg/m(2)) received 6 g AG or placebo 40 min before a 75 g OGTT. The protocol followed the guidelines for the OGTT, with venous blood samples drawn at -40, 0, 15, 30, 45, 60, 90 and 120 min. Ginsenosides in the AG were assessed by established methods for HPLC-UV. RESULTS: Repeated measures analysis of variance demonstrated that there was no significant effect of the AG on incremental plasma glucose (PG) or insulin (PI) or their areas under the curve Indices of insulin sensitivity (insulin sensitivity index (ISI)) and release (deltaPI(30-0)/deltaPG(30-0)) calculated from the OGTT were also unaffected. The AG contained 1.66% total ginsenosides, 0.90% (20S)-protopanaxadiol (PPD) ginsenosides, and 0.75% (20S)-protopanaxatriol (PPT) ginsenosides, with the following key ratios: PPD:PPT of 1.2, Rb(1):Rg(1) of 8.1, and Rb(2):Rc of 0.18. CONCLUSIONS: The present batch of AG was unable to reproduce the postprandial hypoglycemic effects we observed previously. Possible explanations for this discrepancy include marked decrements in total ginsenosides and the key ratios PPD:PPT, Rb(1):Rg(1), and Rb(2):Rc. These data suggest that the ginsenoside profile of AG might play a role in its hypoglycemic effects. The involvement of other components cannot, however, be precluded. PMID- 12571656 TI - Interaction of body mass index and attempt to lose weight in a national sample of US adults: association with reported food and nutrient intake, and biomarkers. AB - OBJECTIVE: This study examined the interaction between body mass index (BMI) and attempting to lose weight for reporting of: (1) macro- and micronutrient intake; (2) intake of low-nutrient-density foods; and (3) serum biomarkers of dietary exposure and cardiovascular disease risk. METHODS: Dietary, anthropometric and biochemical data were from the third National Health and Nutrition Examination Survey (1988-1994), n=13 095. Multiple regression methods were used to examine the independent associations of BMI, trying to lose weight, or the interaction of BMI-trying to lose weight with reported intakes of energy, nutrients, percentage energy from low-nutrient-density foods (sweeteners, baked and dairy desserts, visible fats and salty snacks), and serum concentrations of vitamins, carotenoids and lipids. RESULTS: BMI was an independent positive predictor (P<0.05) of percentage of energy from fat, saturated fat, but a negative predictor of the ratio of reported energy intake to estimated expenditure for basal needs (EI/BEE), percentage of energy from carbohydrate and alcohol (men only), and serum concentrations of folate, vitamin C, vitamin E and most carotenoids in both men and women. Trying to lose weight was a negative predictor (P<0.05) of EI/BEE, intake of energy, and energy density, but not micronutrient intake. Higher mean serum ascorbate, vitamin E, lutein/zeaxanthin, and other carotenoids (men only) concentrations were associated with trying to lose weight (P<0.05) in both men and women. Few adverse BMI-trying to lose weight interaction effects were noted. CONCLUSIONS: There was little evidence of increased nutritional risk in those reportedly trying to lose weight irrespective of weight status. PMID- 12571657 TI - Bone resorption in anorexia nervosa and rehabilitated patients. AB - OBJECTIVE: To assess the impact of anorexia nervosa and that of nutritional rehabilitation on bone resorption. DESIGN: Cross-sectional, observational study. SETTING: Rome, Italy SUBJECTS: Twenty-eight female patients affected by anorexia nervosa (AN, BMIor=18.5 kg/m(2)) and 34 age- and sex-matched healthy controls (CO, BMI >or=18.5 kg/m(2)). Among AN patients, 16 were affected by the 'restrictive' (ANr) and 12 by the 'purging' type (ANp) of anorexia nervosa. METHOD: Body weight, height and skeletal diameters were measured on each individual. The skeletal mass (SKM) was predicted from the skeletal diameters of the elbow, wrist, knee and ankle, using the equation of Martin. Twenty-four-hour urinary excretion of pyridinium crosslinks of collagen (pyridinoline (Pyd) and deoxypyridinoline (Dpd)) and creatinine was assessed by reversed-phase HPLC with fluorimetric detection after solid-phase extraction and by the Jaffe-method with deproteinization, respectively. RESULTS: Twenty-four hour urinary output of Pyd and Dpd was not significantly different between AN and CO when expressed in absolute values, but AN showed higher bone resorption than CO when Pyd and Dpd excretion was adjusted by either creatinine (P<0.0000) or the SKM (P<0.05). Within the AN group, urinary excretion of both cross-links was significantly and consistently higher in ANp compared with ANr (P<0.05). However, these differences disappeared when crosslink output was adjusted either by urinary creatinine or SKM. RE subjects showed no differences in bone resorption with the AN group despite weight gain, being crosslink excretion consistently elevated compared to controls (Pyd: P<0.01 by creatinine and P<0.05 by SKM; Dpd: P<0.01 by creatinine and P<0.05 by SKM). CONCLUSION: Bone resorption is elevated in anorexia nervosa and different strategies for low-weight maintenance do not seem to have a differential impact. Increased bone resorption persists in subjects with past diagnosis of anorexia nervosa despite rehabilitation lasting more than 6 months. This finding indicates that bone mass and turnover should be monitored in anorexia nervosa patients and ex-patients well beyond recovery of normal body mass. Further investigation is warranted to examine the long-term effect of such prolonged increase in bone turnover at a young age. PMID- 12571658 TI - Maternal psychological stress and distress as predictors of low birth weight, prematurity and intrauterine growth retardation. AB - OBJECTIVES: To evaluate: (1) the associations between maternal psychological stress, distress and low birth weight (LBW), prematurity and intrauterine growth retardation (IUGR); (2) the interactions between maternal stress, distress and smoking, alcohol and coffee intake; (3) the prevalences of stress and distress in pregnancy. DESIGN: Longitudinal cohort study. SETTING: Jundiai city, Sao Paulo state, Brazil. SUBJECTS: A total of 865 pregnant women who attended antenatal care between September 1997 and August 2000. METHODS: Measures of stress and distress were obtained, by interview, three times in pregnancy: at a gestational age (GA) lower than 16 weeks, from 20 to 26 weeks and from 30 to 36 weeks. Stress was investigated by the perceived stress scale, PSS, and distress by both the general health questionnaire, GHQ, and the State Trait Anxiety inventories, STAI. The outcomes were: LBW (birth weight <2500 g), prematurity (gestational age (GA) at birth <37 weeks) and IUGR (birth weight for GA 140/90 mmHg or a history of hypertension. Dietary sodium, potassium and calcium intakes were assessed by 24 h recall method or fasting urinary sodium or potassium/creatinine ratios. RESULTS: Seventy-one subjects (64%) were found to have hypertension. Compared with normotensive subjects, hypertensive subjects had lower calcium intake (411+/-s.d. 324 vs 589+/-428 mg, P=0.04), but higher urinary sodium/creatinine ratio (32.6+/-19.3 vs 21.0+/-12.4, P=0.00) and sodium/potassium ratio (4.7+/-2.8 vs 3.4+/-2.3, P=0.02). Among 88 subjects not taking diuretics or antihypertensive drugs, systolic blood pressure was related to calcium intake (r= 0.40), urinary sodium/creatinine ratio (r=0.39), urinary sodium/potassium ratio (r=0.30) and age (r=0.23). Diastolic blood pressure was related to urinary sodium/creatinine (r=0.29). Twenty-three subjects with high urinary sodium/potassium and low calcium intake and 16 subjects with low urinary sodium/potassium ratio and high calcium intake differed markedly with respect to systolic blood pressure (159+/-26 vs 130+/-15 mmHg) and prevalence of hypertension (78% vs 25%). CONCLUSIONS: Older Chinese vegetarians are predisposed to hypertension because of their sodium-rich but calcium-deficient diets. PMID- 12571664 TI - The scored Patient-generated Subjective Global Assessment (PG-SGA) and its association with quality of life in ambulatory patients receiving radiotherapy. AB - OBJECTIVE: To evaluate the scored Patient-generated Subjective Global Assessment (PG-SGA) tool as an outcome measure in clinical nutrition practice and determine its association with quality of life (QoL). DESIGN: A prospective 4 week study assessing the nutritional status and QoL of ambulatory patients receiving radiation therapy to the head, neck, rectal or abdominal area. SETTING: Australian radiation oncology facilities. SUBJECTS: Sixty cancer patients aged 24 85 y. INTERVENTION: Scored PG-SGA questionnaire, subjective global assessment (SGA), QoL (EORTC QLQ-C30 version 3). RESULTS: According to SGA, 65.0% (39) of subjects were well-nourished, 28.3% (17) moderately or suspected of being malnourished and 6.7% (4) severely malnourished. PG-SGA score and global QoL were correlated (r=-0.66, P<0.001) at baseline. There was a decrease in nutritional status according to PG-SGA score (P<0.001) and SGA (P<0.001); and a decrease in global QoL (P<0.001) after 4 weeks of radiotherapy. There was a linear trend for change in PG-SGA score (P<0.001) and change in global QoL (P=0.003) between those patients who improved (5%) maintained (56.7%) or deteriorated (33.3%) in nutritional status according to SGA. There was a correlation between change in PG SGA score and change in QoL after 4 weeks of radiotherapy (r=-0.55, P<0.001). Regression analysis determined that 26% of the variation of change in QoL was explained by change in PG-SGA (P=0.001). CONCLUSION: The scored PG-SGA is a nutrition assessment tool that identifies malnutrition in ambulatory oncology patients receiving radiotherapy and can be used to predict the magnitude of change in QoL. PMID- 12571665 TI - The extent to which breakfast covers the morning energy expenditure of adolescents with varying levels of physical activity. AB - BACKGROUND: Energy intake at breakfast affects the performance of creativity tests, memory recall and voluntary physical endurance in children before lunch, and food craving during the whole day. OBJECTIVES: To assess the adequacy of breakfast energy supply (BES) and energy expenditure (EE) in adolescents during a schoolday without or with 2 h of physical education lesson (PEL) in the morning. DESIGN: Sixty adolescents (four groups of 14-16 boys and girls aged 12-16 y) participated in a cross-sectional study. Activity patterns and EE were determined by whole-body calorimetry during 36 h and in free-living conditions during 5 days using both a diary and the validated heart rate recording method. BES was determined by weighing individual foods. The pyloric energy flow was assessed using a model of fractional stomach emptying. RESULTS: BES averaged 24.9% (s.d.=6.1) of daily EE in the four groups of subjects. It covered the mean morning EE on a schoolday without PEL, but not in a schoolday with 2 h of PEL in any group. When PEL took place from 8-10 am the cumulative EE exceeded the cumulative pyloric energy flow after 105-150 min, that is during the PEL session, and the energy deficit increased until lunch. With a light breakfast (BES-1 s.d.) energy deficiency happened after 90 min. CONCLUSION: The results stress the need for a heavy breakfast for children and adolescents on the days with PEL in the morning, and a carbohydrate rich snack at 10 am to improve attention, memory and willing participation in physical activities. PMID- 12571666 TI - A comparison of the effect of advice to eat either '5-a-day' fruit and vegetables or folic acid-fortified foods on plasma folate and homocysteine. AB - OBJECTIVE: To assess and compare the effects of natural folate (100 micro g) with those of folic acid from fortified sources (100 micro g/day) on plasma folate and homocysteine. DESIGN: Randomized controlled trial (parallel groups). SETTING: Men and women living in South Wales, UK. SUBJECTS: A total of 135 healthy individuals recruited from the local workforce and blood donor sessions. All subjects possessed the 'wild-type' CC genotype for C677T polymorphism in methylenetetrahydrofolate reductase (MTHFR). INTERVENTIONS: Subjects underwent one of the following dietary interventions for 4 months: (1) fortified diet-usual diet plus 100 microg/day folic acid from fortified foods; (2) natural folate diet usual diet plus 100 microg/day folate from natural sources; (3) control-usual diet. RESULTS: The fortified group increased reported intake of folic acid from fortified foods compared to other groups (P<0.001) achieving an extra 98 microg/day (95% CI 88-108). The natural folate group increased reported intake of natural source folates compared with the other two groups (P<0.001), but achieved a mean increase of only 50 microg/day (95% CI 34-66). Plasma folate increased (P<0.01) by a similar amount in both intervention groups compared to controls (fortified group 2.97, 95% CI 0.8-5.1; natural group 2.76, 95% CI 0.6-4.9. Plasma homocysteine, vitamins B(6) and B(12) were not significantly changed. CONCLUSIONS: Subjects achieved increases in folate intake using fortified foods more easily than by folate-rich foods, however both sources increased plasma folate by a similar amount. These levels of intake were insufficient to reduce homocysteine concentrations in MTHFR CC homozygotes, but may be more effective in other genotypes. PMID- 12571667 TI - A randomized controlled trial of phytoestrogen supplementation, growth and bone turnover in adolescent males. AB - OBJECTIVE: To assess the effect of phytoestrogens on bone turnover and growth in adolescent boys. DESIGN: Randomized double-blind placebo-controlled trial. SETTING: Single school in northwest Tasmania. PARTICIPANTS: Adolescent boys (treatment n=69, placebo n=59, mean age 16.8 y). INTERVENTIONS: Six weeks of isoflavone supplementation (Novasoy, 50 mg daily of isoflavone equivalents). Bone turnover markers (bone specific alkaline phosphatase (BAP) and pyridinoline creatinine ratio (PYR)) were measured at baseline and follow-up. RESULTS: Despite marked increases in urinary genistein and daidzein in the treatment arm (both P<0.001), there were no significant differences in BAP, PYR or short-term height or weight change. This applied to both intention-to-treat and per protocol analysis. Neither was there a significant correlation between urinary genistein and daidzein levels and BAP or PYR. CONCLUSIONS: Phytoestrogen supplementation to the level of usual Japanese dietary intake has no measurable effect on bone turnover in adolescent boys. Longer-term studies of bone density may be desirable but it is unlikely that there will be a large effect in either girls or boys given the lower endogenous oestrogen levels in boys. PMID- 12571668 TI - Nutrient intake in women with primary and secondary Sjogren's syndrome. AB - OBJECTIVE: Recently, it has been proposed that dietary factors may contribute to the etiology and progression of Sjogren's syndrome, and that nutritional intervention may modify the severity of pathological abnormalities. The objective of this study was to determine whether the nutrient intake of women with primary (1 degrees SS) or secondary (2 degrees SS; ie with systemic lupus erythematosus (2 degrees SS/SLE) or rheumatoid arthritis (2 degrees SS/RA) Sjogren's syndrome is significantly different than that of age- and gender-matched controls. DESIGN: Women with Sjogren's syndrome were asked to complete the 97 General Purpose Semi Quantitative Food Frequency Questionnaire, which consists of a list of 147 separate food items that represent the major sources of multiple nutrients. Nutritional data were evaluated in terms of absolute and energy-adjusted nutrient amounts and analyzed by ANOVA. RESULTS: Our results showed: (a) greater intake of energy, glutamate, carbohydrates, lactose, phosphorus, caffeine and unsupplemental thiamin and riboflavin in 1 degrees SS, as well as supplemental calcium in 2 degrees SS/SLE, compared with controls; (b) greater nutrient intake of energy, protein, glutamate, methionine, tryptophan, carbohydrates, lactose, supplemental calcium and phosphorus, sodium, caffeine and unsupplemental calcium, riboflavin and thiamin in 2 degrees SS/RA, relative to 1 degrees SS and/or 2 degrees SS/SLE; (c) higher energy-adjusted values for supplemental calcium in 2 degrees SS/SLE, and for vitamin A and supplemental iron and zinc in 2 degrees SS/RA, compared with other groups; and (d) higher energy-adjusted intake of supplemental calcium, and a lower energy-adjusted intake of unsupplemental vitamin C, polyunsaturated fat, linoleic acid, omega-3 fatty acid, and specific other unsaturated fatty acids, in the Sjogren's syndrome group as a whole, relative to controls. CONCLUSIONS: Our findings demonstrate that nutrient intake is altered in Sjogren's syndrome. PMID- 12571669 TI - Predicting basal metabolic rate in the obese is difficult. AB - OBJECTIVE: To reassess the validity of the Schofield equations for predicting basal metabolic rate (BMR) in the obese. DATA: The data collection compiled from many studies by Schofield in 1985. METHODS: Nonparametric curve fitting and comparison of groups. RESULTS: BMR increases more slowly at heavier weights in men (above about 75 kg) and women (above about 65 kg), and to ignore this is to predict overestimates of BMR. Data obtained from studies of different populations show substantial differences, which are influential when data are combined. CONCLUSIONS: The Schofield equations are unsuitable for obese populations. Given that current Western populations exhibit prevalences of obesity many times greater than those in the Schofield database, this indicates the need for further study of suitable predictors for these individuals. PMID- 12571670 TI - Modifying children's food preferences: the effects of exposure and reward on acceptance of an unfamiliar vegetable. AB - OBJECTIVE: The aim of this study was to evaluate two interventions (one reward based and one exposure-based) for increasing children's acceptance of an unfamiliar vegetable compared with a no-treatment control. It was predicted that the exposure condition would increase liking for, and consumption of, the vegetable relative to either the reward or control group. DESIGN: Using a randomized controlled design, participants were assigned to one of two intervention groups (exposure or reward) or to a no-treatment control condition, for a 2 week period. Liking for, and consumption of, red pepper was assessed before and after the treatment period. SETTING: The study was conducted in three primary schools in London. SUBJECTS: Parental consent was obtained for 49 out of a possible 72 children. INTERVENTIONS: Interventions comprised eight daily sessions during which participants in the exposure group were offered a taste of sweet red pepper and told that they could eat as much as they liked. Participants in the reward group were shown a sheet of cartoon stickers and told that they could choose one of them on condition that they ate at least one piece of the pepper. RESULTS: The exposure-based intervention significantly increased both liking (P=0.006) and consumption (P=0.03) compared with the control group. The outcome of the reward intervention was intermediate and did not differ significantly from the exposure or control conditions. CONCLUSIONS: Repeated exposure to the taste of unfamiliar foods is a promising strategy for promoting liking of previously rejected foods in children. PMID- 12571671 TI - Relationship between methylmalonic acid, homocysteine, vitamin B12 intake and status and socio-economic indices, in a subset of participants in the British National Diet and Nutrition Survey of people aged 65 y and over. AB - OBJECTIVE: Assessment of functional vitamin B(12) status in a subset of the respondents in the British National Diet and Nutrition Survey of people aged 65 y and over. SETTING: National Diet and Nutrition Survey: a British nationwide cross sectional sample of people aged 65 y and over, living either in the community or in institutions such as nursing homes, during one calendar year spanning 1994 1995. METHODS: Methylmalonic acid (MMA) concentrations were measured in plasma samples from 313 subjects (ca 14% of those originally enrolled in the survey). The results were compared with those for serum vitamin B(12), vitamin B(12) intakes and other status and intake estimates and with socio-demographic indices. RESULTS: Of the NDNS participants overall, 20% had serum vitamin B(12) concentrations<150 pmol/l. In the subset studied here, 24% of free-living and 46% of institution-living participants had MMA>0.5 micromol/l. Geometric mean MMA increased with age, from 0.25 micro mol/l in people aged 65-74 y to 0.38 micro mol/l in people aged 85+y. There was little evidence for any gender difference in MMA. It was inversely correlated with serum vitamin B(12) and with red blood cell folate; it was positively correlated directly with total homocysteine, but not significantly with serum folate or with vitamin B(12) intake. Among respondents with high MMA, a subgroup had normal serum vitamin B(12) but higher-than-average plasma urea and creatinine. Socio-demographic co-variates of MMA included receipt of State income benefits, social class of head of household, and educational attainment. These indices were not correlated with serum vitamin B(12). CONCLUSIONS: The progressive increase in MMA with age is metabolic evidence for increasing risk of functional vitamin B(12) deficiency with increasing age in older people. There is evidence that renal function is linked to high MMA in some older people. Age and renal function are thus both important when establishing upper reference limits for MMA. The socio-demographic observations suggest a link between poverty and poor functional vitamin B(12) status in older British people. PMID- 12571672 TI - Implications of the prevalence of stunting, overweight and obesity amongst South African primary school children: a possible nutritional transition? AB - OBJECTIVE: To investigate the relationship between stunting and levels of overweight/obesity among South African school children, using two definitions of overweight and obesity, based on the WHO and International Obesity Task Force (IOTF) criteria. DESIGN: Cross-sectional descriptive analysis of the nutritional status of primary school children, using primary data from a rural community based study undertaken in 1995 and secondary data from the South African National Primary Schools (SANPS) survey conducted in 1994. Stunting was measured according to the WHO definition of -2 Z scores height-for-age. Two sets of criteria were used to measure overweight and obesity-the WHO/NCHS standard based on the 85th and 95th centiles and the IOTF criteria. SETTING: The primary data source was from a rural KwaZulu-Natal community based survey. The secondary data source SANPS consisted of data at National and Provincial level; for this study only data from the province of KwaZulu-Natal was considered. SUBJECTS: Primary school children aged between 8 and 11 y of age; 802 from the primary data source and 24 391 from the secondary source. RESULTS: Moderate stunting ranged from 2.9 to 40.2%, and mild stunting ranged from 31.4 to 75%. The prevalence of overweight ranged from 0.4 to 13.3% (WHO criteria) and from 0.4 to 11.9% using the IOTF criteria; while obesity ranged from 0.1 to 3.7% (WHO) and from 0.1 to 1.5% (IOTF criteria). The prevalence of overweight and obesity was observed to be higher using the WHO definition than that of IOTF (0.050.55, in all cases). The levels of agreement in all cases were less for obesity than overweight (both ranged from 0.55 to 1.0). Females were observed to have higher kappa levels than their male counterparts; they also had higher prevalence levels of overweight and obesity across age and geographical group. Uniformly high levels of both mild and moderate stunting were observed both nationally and provincially. However, no excess relative risk of being overweight if stunted was observed in this study (P>0.05). CONCLUSIONS: Caution must be applied when using either definition for obesity. However, very high levels of agreement occur for overweight. There is no obvious relationship between overweight and stunting in this study, but high levels of mild stunting were observed. Regular clinical and epidemiological monitoring of nutritional status needs to be undertaken in South Africa to examine possible future trends of overweight/obesity and their relationship with stunting, and for comparisons with global trends. PMID- 12571673 TI - Estimated energy intake, macronutrient intake and meal pattern of Flemish adolescents. AB - OBJECTIVE: To describe the energy and macronutrient intake and the meal patterns of Flemish adolescents, aged 13-18 y. METHODS: A 7 day estimated food record was administered to the whole sample. SETTING: Secondary schools in the city of Ghent, Belgium. SUBJECTS: A total of 341 adolescents (13-18 y) selected by a multistage clustered sampling (participation: 72.7%). MAIN RESULTS: A significant increase with age was observed in total energy intake in adolescent boys (P<0.01), but not in girls. The energy distribution over the macronutrients showed no significant difference between boys and girls. On average, 35.7% (s.d. 4.81%) of energy came from total fat and 15.4% (s.d. 2.46%) from saturated fatty acids; 49.0% (s.d. 5.28%) from total carbohydrates with 25.1% (s.d. 4.49%) from complex carbohydrates and 23.9% (s.d. 5.86%) from free sugars. The energy contribution of alcohol in the 16-18 y-old-group was significantly higher as compared with the 13-15 y-old-group, for both boys and girls. Snacks between meals accounted for almost 20% of the total energy intake. Lunch and dinner were characterized by high total fat content. CONCLUSION: These students consumed a diet high in total fat and in saturated fatty acids and also high in mono- and disaccharides. Observed mean intakes deviate considerably from the Belgian dietary guidelines. A low energy intake at breakfast was observed, while a higher proportion of energy was derived from snacks. PMID- 12571674 TI - Effect of high vegetable protein diets on urinary calcium loss in middle-aged men and women. AB - OBJECTIVE: To determine the effect of high-protein diets, which have recently been promoted for their health benefits, on urinary calcium losses and bone turnover in older subjects. DESIGN: Randomized controlled cross-over study. SETTING: Teaching hospital and university. SUBJECTS: Twenty hyperlipidemic men and postmenopausal women (age 56+/-2 y) completed the study. INTERVENTION: One month test and control phases during which subjects consumed equi-energy metabolic diets high in calcium (1578 and 1593 mg/day, respectively). On the test diet 11% of total dietary energy from starch in the control bread was replaced by protein (wheat gluten), resulting in 27% of energy from protein on the test diet vs 16% on the control diet. MAIN OUTCOME MEASURE: Urinary calcium excretion. RESULTS: Compared with the control diet, at week 4, the test diet increased mean (+/-s.e.m.) 24 h urinary output of calcium (139+/-15 vs 227+/-21 mg, P=0.004). The treatment difference in urinary calcium loss correlated with the serum anion gap as a marker of metabolic acid production (r=0.57, P=0.011). Serum calcium levels were marginally lower 2.41+/-0.02 vs 2.38+/-0.02 mmol/l (P=0.075), but there was no significant treatment difference in calcium balance, possibly related to the high background calcium intake on both diets. CONCLUSION: In the presence of high dietary calcium intakes the vegetable protein gluten does not appear to have a negative effect on calcium balance despite increased urinary calcium loss. PMID- 12571675 TI - Effect of diet on plasma total antioxidant status in phenylketonuric patients. AB - BACKGROUND: Phenylketonuria (PKU), an inborn error of phenylalanine (Phe) metabolism, is treated with a low Phe lifelong diet, which is a vegetarian and contains many antioxidants. AIM: The aim of this study was to evaluate the effect of diet on plasma total antioxidant status in our PKU patients. METHODS, RESULTS: Twenty-two PKU patients on strict diet (group A), 24 PKU patients who did not accurately follow their dietary control (group B) and 40 healthy children (controls) of comparable age took part in this study. Nutrients, as well as blood levels of lipids, vitamin C, beta-carotene and alpha-tocopherol were evaluated in all groups. Vitamin C intake and its blood levels did not differ between the groups. However, the intake of beta-carotene, alpha-tocopherol (2211+/-116, 14+/ 1.0 mg/24 h) and their blood levels (0.7+/-0.09, 34+/-0.9 micro mol/l) in group A were statistically significantly higher (P<0.001) as compared with those of group B (1352+/-118, 10+/-1.0 mg/24 h and 0.49+/-0.08, 22+/-0.6 micromol/l) and controls (1290+/-120, 9.0+/-0.9 mg/24 h and 0.40+/-0.09, 24+/-1.6 micromol/l). Lipid intakes and their blood levels were lower in patients on the strict diet. Plasma total antioxidant status was higher in the same group of patients (group A). Additionally, positive correlations were found between the antioxidant vitamin blood levels and the plasma total antioxidant status in the groups, especially in the group A. PKU patients of group A showed significantly higher antioxidant status (1.6+/-0.2 mmol/l) as compared with those of group B (1.0+/ 0.19 mmol/l; P<0.001) and controls (1.01+/-0.2 mmol/l). CONCLUSIONS: The high plasma antioxidant status in patients with PKU, especially in those with a good compliance with their diet, is possibly due to the amounts of antioxidants which are present in their special low Phe vegetarian diet. PMID- 12571676 TI - NTP Toxicology and Carcinogenesis Studies of Polyvinyl Alcohol (CAS No.9002-89-5) in Female B6C3F1 Mice (Intravaginal Studies). AB - Polyvinyl alcohol is produced primarily for use in textile sizing, adhesives, polymerization aids, and paper coatings. It is also used in surgical drapes, towels, and gauze sponges; protective gloves; cosmetic formulations; topical ophthalmic preparations; plastic sponge implants for reconstructive surgery; and intravaginal contraceptive foam and film. In addition, polyvinyl alcohol is used with magnesium sulfate to dilate the cervix of women prior to induction of labor. It is estimated that hundreds of thousands of women in the United States use an intravaginal product containing polyvinyl alcohol each year. The Food and Drug Administration nominated low-viscosity polyvinyl alcohol for a 2-year study because of concern about the lack of information about the long-term toxic and carcinogenic effects by the intravaginal route. Female B6C3F1 mice received polyvinyl alcohol (approximately 99% pure) in deionized water by intravaginal administration for 30 days or 2 years. 30-DAY STUDY IN MICE: Three groups of 50 female B6C3F1 mice were used in this intravaginal study. The vehicle control group received only 20 &mgr;L of a deionized water vehicle. The other two groups each received 20 &mgr;L of 25% polyvinyl alcohol in deionized water. Animals in one dose group were returned to their cages after dosing; animals in the other dose group were restrained in a vertical nose-down position in restraint bags for several minutes after dosing. Animals were dosed daily for 30 consecutive days. All mice survived to the end of the study. The final mean body weights and body weight gains of dosed mice were similar to those of the vehicle control group. Abnormalities noted in the vaginal area after dosing included vaginal plugs, secretions, and swelling. These vaginal changes were minimal to mild and occurred in vehicle controls as well as in dosed mice. Restraint of mice after dosing appeared to eliminate vaginal secretions but increased both the incidence of vaginal irritation and the severity of vaginal opening swelling. At necropsy, mildly enlarged uterine horns were observed in 10 vehicle control mice, three 25% mice, and seven 25% (restrained) mice. No chemical-related lesions were observed. 2-YEAR STUDY IN MICE: Three groups of 100 female B6C3F1 mice were used in this intravaginal study: an untreated control group, a vehicle control group receiving 20 &mgr;L deionized water vehicle only, and a dosed group receiving 20 &mgr;L 25%% polyvinyl alcohol in deionized water. Animals were dosed 5 days per week, excluding holidays, for 104 to 105 weeks. Survival, Body Weights, and Clinical Findings: Survival of dosed mice was similar to that of the two control groups. The final mean body weight of vehicle control mice was less than that of the untreated control group. The mean body weights of the dosed mice were less than those of the untreated controls from week 17 until the end of the study. The only clinical finding was vaginal irritation, observed in six mice in the vehicle control group and 11 mice in the dosed group. Pathology Findings: No neoplasms or nonneoplastic lesions related to chemical treatment were observed. The incidences of reproductive tract nonneoplastic lesions in the dosed group did not differ significantly from those in the vehicle control group; similarly, the incidences of reproductive tract nonneoplastic lesions in the vehicle control group did not differ significantly from those in the untreated control group. CONCLUSIONS: Under the conditions of this 2-year study, there was no evidence of carcinogenic activity of polyvinyl alcohol (molecular weight approximately 24,000) in female B6C3F1 mice administered 20 &mgr;L of a 25%% solution intravaginally. There were no neoplasms or nonneoplastic lesions considered related to treatment with polyvinyl alcohol. Synonyms: Ethenol homopolymer, PVA Trade names: Akwa Tears, Alcotex, Alvyl, Aracet, Cipoviol, Covol, Elvanol, Ethenol, Gelvatol, Gohsenol, Ivalon, Kuralon, Kurare, Lemol, Liquifilm, Mowiol, Polydesis, Polysizer, Polyvinol, Polyviol, Poval, Resistoflex, Rhodoviol, Sno, Poval, Resistoflex, Rhodoviol, Sno Tears, Solvar, Sumitex, Vibatex, Vinacol, Vinalak, Vinarol, Vinarole, Vinavilol, Vinol, Vinylon PMID- 12571677 TI - NTP Toxicology and Carcinogenesis Studies of Theophylline (CAS No. 58-55-9) in F344/N Rats and B6C3F1 Mice (Feed and Gavage Studies). AB - Theophylline is an alkaloid found in tea (Thea sinensis) and chocolate and is structurally related to caffeine and theobromine. Theophylline is used as a pharmaceutical agent. It stimulates the heart and central nervous system, relaxes the smooth muscles of the bronchi and blood vessels, and causes diuresis. The drug is used mainly as a bronchodilator in obstructive airway diseases, such as bronchial asthma, and for myocardial stimulation. Theophylline was nominated for toxicologic and carcinogenicity testing as a representative of the purine structural subclass, particularly because of its relationship to purines such as caffeine, 1-methyl-3-hydroxyguanine, and 3-hydroxy-1-methylxanthine, the latter two compounds having been shown to induce sarcomas in rats. Additional reasons for testing theophylline included its widespread use in humans as a pharmaceutical agent, its possible genotoxicity in vitro, and the lack of information on its potential toxicity and/or carcinogenicity under conditions of chronic oral usage. Based on reported teratogenicity and testicular toxicity, it was also recommended that reproductive studies be included in the evaluation of theophylline. The oral route of administration was selected because it is the primary route of human exposure, and the gavage route was selected because it mimics the pharmaceutical use of theophylline in humans. Male and female F344/N rats and B6C3F1 mice were given theophylline (greater than 99% pure) in feed or in corn oil by gavage for 16 days or 14 weeks or in corn oil by gavage for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, mouse bone marrow, and mouse peripheral blood. 16-DAY FEED STUDY IN RATS: Groups of five male and five female F344/N rats were given 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm theophylline in feed for 16 days, which resulted in approximate daily doses of 50, 100, 250, 450, or 1,000 mg theophylline/kg body weight to males and 75, 150, 250, 450, or 1,100 mg/kg to females. All rats survived until the end of the study. The final mean body weights and body weight gains of 8,000 ppm males and females were significantly less than those of the controls. The absolute and relative testis weights of 4,000 ppm males were significantly greater than those of the controls. Increased incidences of uterine hypoplasia were observed microscopically in exposed groups of females. 16-DAY GAVAGE STUDY IN RATS: Groups of five male and five female F344/N rats were given 0, 12.5 (twice daily), 25 (once daily), 50 (once daily), 50 (twice daily), 100 (once daily), 200 (once daily), 200 (twice daily), or 400 (once daily) mg theophylline/kg body weight in corn oil by gavage. All rats receiving 400 mg/kg once daily and all but one female receiving 200 mg/kg twice daily died during the study. In groups dosed once daily, final mean body weights and body weight gains of males receiving 100 or 200 mg/kg and mean body weight gains of females receiving 50, 100, or 200 mg/kg were less than those of controls. The final mean body weights and body weight gains of groups receiving theophylline twice daily were generally similar to those of groups receiving the same daily dosages once daily. Clinical findings included rapid or labored respiration, hunched posture, and squinting. In groups dosed once daily, absolute and relative uterus weights of females receiving 100 or 200 mg/kg once daily were significantly less than those of the controls, and the absolute and relative uterus weights of females receiving 100 mg/kg once daily were significantly less than those of females receiving 50 mg/kg twice daily. Uterine atrophy was observed in three females receiving 200 mg/kg twice daily. Periarteritis of the mesenteric arteries was observed in two males and two females receiving 400 mg/kg once daily. 16-DAY FEED STUDY IN MICE: Groups of five male and five female B6C3F1 mice were given 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm theophylline in feed for 16 days, resulting in approximate daily doses of 250, 475, 950, 1,800, or800, or 2,000 mg theophylline/kg body weight to males and 300, 450, 1,225, 2,000, or 4,375 mg/kg to females. All mice survived until the end of the study. Final mean body weights of 4,000 and 8,000 ppm females and mean body weight gains of 2,000, 4,000, and 8,000 ppm females were significantly greater than those of the controls. Feed consumption by exposed groups was similar to that by the controls, except that by the 8,000 ppm males, which was approximately 40% the amount of feed consumed by the control group. Histopathologic examinations were not performed due to the absence of mortality and significant exposure-related lesions. 16-DAY GAVAGE STUDY IN MICE: Groups of five male and five female B6C3F1 mice were given 0, 12.5 (twice daily), 25 (once daily), 50 (once daily), 50 (twice daily), 100 (once daily), 200 (once daily), 200 (twice daily), or 400 (once daily) mg theophylline/kg body weight in corn oil by gavage. Three males and all females receiving 400 mg/kg once daily died on day 1. There were no significant differences in final mean body weights or body weight gains. There were no histopathologic findings attributed directly to theophylline. 14-WEEK FEED STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were given 0, 1,000, 2,000, or 4,000 ppm theophylline in feed for 14 weeks, which resulted in approximate daily doses of 75, 125, or 250 mg theophylline/kg body weight to males and 75, 125, or 275 mg/kg to females. The final mean body weight of 1,000 ppm females was significantly greater than that of the control group. Feed consumption by exposed groups was similar to that by the controls. Mean cell volume and mean cell hemoglobin were significantly greater in males exposed to 2,000 or 4,000 ppm than those in the control group. Segmented neutrophil counts of all groups of exposed females were significantly greater than that of the control group. The absolute and relative kidney weights of 4,000 ppm males were significantly greater than those of the controls, and there was an exposure related increase in the severity of nephropathy in males. Exposure-related increases in the incidences of mesenteric and/or pancreatic periarteritis were observed in males and females. 14-WEEK GAVAGE STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were given 0, 37.5, 75, or 150 mg theophylline/kg body weight in corn oil by gavage for 14 weeks. One male and one female receiving 150 mg/kg died before the end of the study. The mean body weight gain of 150 mg/kg females was significantly greater than that of the controls. Mean cell volume of 150 mg/kg males and mean cell hemoglobin of all groups of dosed males were significantly greater than those of the control group. There were slight dose dependent increases in the incidences of mesenteric periarteritis in dosed males and females. 14-WEEK FEED STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were given 0, 1,000, 2,000, or 4,000 ppm theophylline in feed for 14 weeks, resulting in approximate daily doses of 175, 400, or 800 mg theophylline/kg body weight to males and 225, 425, or 850 mg/kg to females. All mice survived until the end of the study. The final mean body weights and body weight gains of all exposed groups of males and females were significantly less than those of the controls. Feed consumption by exposed groups was similar to that by the controls. Leukocyte, segmented neutrophil, and lymphocyte counts of 4,000 ppm males were significantly greater than those of the controls. Leukocyte and segmented neutrophil counts of 2,000 or 4,000 ppm females were significantly greater than those of the controls. There were no histopathologic findings attributed directly to theophylline exposure. 14-WEEK GAVAGE STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were given 0, 75, 150, or 300 mg theophylline/kg body weight in corn oil by gavage for 14 weeks. Three males and all females receiving 300 mg/kg, one 75 mg/kg male, and one control female died before the end of the study. Final mean body weights and body weight gains of 150 and 300 mg/kg males were significantly less than those of the controls. Mean cell volume and mean cell hemoglobin of 300 mg/kg males were significantly greater than those of the controls. There were no histopathologic findings attributed directly to theophylline treatment. 2-YEAR GAVAGE STUDY IN RATS: Groups of 50 male and 50 female rats were given 7.5, 25, or 75 mg theophylline/kg body weight in corn oil by gavage for 2 years. Survival and Body Weights: There were no significant differences in survival between dosed and control groups. Final mean body weights of all groups of dosed males and females were significantly less than those of the controls. Pathology Findings: There were no significantly increased incidences of neoplasms in dosed rats. The incidence of chronic inflammation of the mesenteric arteries was significantly increased in males receiving 75 mg/kg compared to the controls. There were doserelated negative trends in the incidences of mammary gland fibroadenoma and fibroadenoma or carcinoma (combined) in females; these differences correlated with decreased body weights. 2-YEAR GAVAGE STUDY IN MICE: Groups of 50 male B6C3F1 mice were given 0, 15, 50, or 150 mg theophylline/kg body weight and groups of 50 female B6C3F1 mice were given 0, 7.5, 25, or 75 mg/kg in corn oil by gavage for 2 years. Survival and Body Weights: Survival of 150 mg/kg males was significantly less than that of the controls. The final mean body weights of 150 mg/kg males, 25 mg/kg females, and 75 mg/kg females were significantly less than those of the control groups. Pathology Findings: There were no treatment-related increases in incidences of nonneoplastic lesions or neoplasms. In males and females, there were decreased incidences of hepatocellular adenoma and of the combined incidences of hepatocellular adenoma or carcinoma compared to the controls. Male mice had a pattern of nonneoplastic liver lesions along with silver-staining helical organisms in the liver consistent with Helicobacter hepaticus infection. The incidences of these liver lesions in 150 mg/kg males were significantly lower than those in control males. Increases in the incidences of hepatocellular neoplasms in male mice have been shown to be associated with H. hepaticus infection when hepatitis is also present. Because of this association, interpretation of the decreased incidence of liver neoplasms in male mice was more difficult. Incidences of lesions at other sites in this study were not considered to have been significantly impacted by H. hepaticus infection or its associated hepatitis. GENETIC TOXICOLOGY: Theophylline was not mutagenic in Salmonella typhimurium, with or without metabolic activation (S9). It induced sister chromatid exchanges but not chromosomal aberrations in cultured Chinese hamster ovary cells. The positive sister chromatid exchange response was noted only in the absence of S9. In vivo, a mouse bone marrow sister chromatid exchange test showed positive results at a standard 23-hour harvest time; however, this test was not repeated and the response is unconfirmed. An in vivo mouse bone marrow chromosomal aberrations test, that employed both standard and extended exposure protocols, gave negative results. The frequency of micronucleated erythrocytes was determined in peripheral blood of male and female mice exposed to theophylline in dosed feed or in corn oil by gavage for 14 weeks. No significant increases in the frequencies of micronucleated cells were seen in male or female mice in either of the studies. CONCLUSIONS: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity of theophylline in male or female F344/N rats administered 7.5, 25, or 75 mg/kg. There was no evidence of carcinogenic activity of theophylline in male B6C3F1 mice administered 15, 50, or 150 mg/kg or female B6C3F1 mice administered 7.5, 25, or 75 mg/kg. Gavage administration of theophylline caused chronic inflammation of the mesenteric arteries in dosed male rats. Decreased incidences of mammary neoplasms in female rats were likely associated with lower body weights. There were dose-related decreases in the incidences of hepatocellular adenoma and hepatocellular carcinoma in male and female mice. Synonyms: 3,7 dihydro-1,3-dimethyl-1H-purine-2,6-dione; 1,3-dimethylxanthine; 1H-purine-2,6 dione; NSC 2066; pseusdotheophylline; theocin; theophyllin; theophylline, anhydrous Trade names: Accurbron; Aerobin; Aerolate III; Afonilum; Aminophylline; Aquaphyllin; Armophylline; Asmalix; Bilordyl; Bronchoretard; Bronkodyl; Cetraphylline; Constant-T; Diffumal; Duraphyl; Duraphyllin; Elixicon; Elixophyllin; Euphylline L.A.; Euphylong; LaBID; Labophylline; Lanophyllin; Lasma; Liquophylline; Optiphyllin; Parkophyllin; Phylocontin; Physpan; Pro-Vent; PulmiDur; Pulmo-Timelets; Quibron; Respbid; Rona-Phyllin; Sabidal; Slo-bid; Slo Phyllin; Solosin; Sustaire; Tefamin; Teobid; Teofyllamin; Tesona; Theal tablets; Theo-24; Theobid; Theocap; Theochron; Theoclear; Theocontin; Theo-Dur; Theofol; Theograd; Theolair; Theolan; Theolix; Theophyl; Theoplus; Theo-Sav; Theosol; Theospan; Theostat; Theovent; TheoX; T-Phyl; Truphylline; Uni-Dur; Unifyl; Uniphyl; Uniphyllin; Xanthium PMID- 12571678 TI - NTP Toxicology and Carcinogenesis Studies of Oxymetholone (CAS NO. 434-07-1) in F344/N Rats and Toxicology Studies of Oxymetholone in B6C3F1 Mice (Gavage Studies). AB - Oxymetholone is a synthetic anabolic steroid used to treat a variety of conditions, including hypogonadism and delayed puberty. It is also used to correct hereditary angioneurotic edema, manage carcinoma of the breast, promote a positive nitrogen balance following injury or surgery, and stimulate erythropoiesis. Considerable amounts of androgens are consumed by athletes in attempts to improve athletic performance. The National Institute of Environmental Health Sciences and the National Cancer Institute nominated oxymetholone for study based on its extensive illicit pharmaceutical use and the limited evidence that it is a potential human carcinogen. Male and female F344/N rats received oxymetholone (greater than 99% pure) in 0.5% methylcellulose by gavage for 16 days, 14 weeks, or 2 years, and male and female B6C3F1 mice received oxymetholone in 0.5% methylcellulose by gavage for 16 days or 14 weeks. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and mouse peripheral blood erythrocytes. 16-DAY STUDY IN RATS: Groups of five male and five female F344/N rats were administered 0, 160, 315, 625, 1,250, or 2,500 mg oxymetholone/kg body weight in 0.5% methylcellulose by gavage for 16 days. All male rats survived to the end of the study; one 2,500 mg/kg female died on day 14. The mean body weights of all dosed groups of males were significantly less than those of the vehicle controls, while those of 160 and 315 mg/kg females were significantly greater. 16-DAY STUDY IN MICE: Groups of five male and five female B6C3F1 mice were administered 0, 320, 630, 1,250, 2,500, or 5,000 mg/kg in 0.5% methylcellulose by gavage for 16 days. All mice survived to the end of the study. The final mean body weights of all dosed groups of females were greater than those of the vehicle controls. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were administered 0, 80, 160, 315, 625, or 1,250 mg/kg in 0.5% methylcellulose by gavage for 14 weeks. One male rat each in the 625 and 1,250 mg/kg groups died before the end of the study. The mean body weights of males administered 160 mg/kg or greater were significantly less than those of the vehicle controls; in contrast, the mean body weights of all dosed groups of females were significantly greater. A dose-related erythrocytosis, evidenced by increases in erythrocyte counts, total hemoglobin concentrations, and hematocrit values, occurred in dosed groups of rats at week 14. A dose-related hypocholesterolemia occurred at all time points in all dosed groups of rats. Dose and time-related decreases in 5 -nucleotidase activity occurred in treated rats. There was a transient, treatment-related increase in the activity of alanine aminotransferase in males and females. For male rats administered oxymetholone, cauda epididymis, epididymis, and testis weights and spermatid counts and total spermatid heads per testis were significantly less than those of the vehicle controls, and total spermatid heads per gram testis were significantly greater. Female rats in the 80 mg/kg group spent more time in diestrus and less time in estrus than did the vehicle controls. Kidney weights of males and females and liver and uterus weights of females were increased compared to vehicle controls in rats that received 315 mg/kg or greater; thymus weights of males and females and sartorius muscle and testis weights of males were less. Compared to the vehicle controls, rats that received 160 mg/kg or greater had increased incidences of nonneoplastic lesions of the kidney and mammary gland, and the incidences of hydrometra of the uterus and dysgenesis of the ovary were increased in dosed groups of females. Female rats administered 315 mg/kg or greater had increased incidences of cytoplasmic vacuolization of the adrenal gland and myocardial degeneration of the heart. The severities of these lesions generally increased with increasing dose. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were administered 0, 160, 320, 630, 1,250, or 2,500 mg/kged 0, 160, 320, 630, 1,250, or 2,500 mg/kg in 0.5% methylcellulose by gavage for 14 weeks. All mice administered oxymetholone survived until the end of the study. The mean body weights of all dosed groups were similar to those of the vehicle controls. The percentages of motile sperm in 1,250 and 2,500 mg/kg males were significantly less than those of the vehicle controls. The estrous cycle lengths of 630, 1,250, and 2,500 mg/kg females were significantly longer, and females in the 1,250 and 2,500 mg/kg groups spent more time in diestrus and less time in estrus. Kidney and liver weights of males and females were greater and thymus weights of females were less than those of the vehicle controls. All dosed females had hyperplasia of the clitoral gland, metaplasia of the parietal layer epithelium of the Bowman's capsule in the kidney, and cytoplasmic alteration of the submandibular gland; these lesions were not observed in the vehicle control group. The incidences of hypoplasia of the ovary in 320 mg/kg or greater females and of parotid gland atrophy in 1,250 and 2,500 mg/kg females were increased. The results of the 14-week oral gavage studies were generally similar in rats and mice, but rats were much more sensitive to oxymetholone. Because it was not likely that a long-term mouse study would provide significant additional toxicity information, the NTP decided to conduct a 2-year study in rats only. 2-YEAR STUDY IN RATS: Groups of 90 male F344/N rats were administered 0, 3, 30, or 150 mg/kg in 0.5% methylcellulose by gavage, and 90 female F344/N rats were administered 0, 3, 30, or 100 mg/kg in 0.5% methylcellulose by gavage for up to 104 weeks, with 9 or 10 rats per group evaluated at 3, 6, 12, or 18 months. Survival and Body Weights: Survival of all dosed groups was similar to that of the vehicle controls. The mean body weights of the 30 mg/kg male group were generally within 10% of those of the vehicle controls, but those of the 150 mg/kg group were markedly decreased. Mean body weights of 3 and 30 mg/kg females were generally greater than those of the vehicle controls throughout the study. Determinations of Oxymetholone in Plasma: The concentrations of oxymetholone in plasma of male and female rats receiving 3 mg/kg for 6, 12, or 18 months were generally below the limits of quantification; therefore, all plasma concentrations in the 3 mg/kg group are considered to be estimates (Table 8). The plasma concentrations at 30 mg/kg were approximately one order of magnitude greater than those of the estimates for males and females receiving 3 mg/kg. There were no dose-related differences in plasma concentrations in female rats receiving 30 or 100 mg/kg, but plasma concentrations in males were significantly elevated in the 150 mg/kg group. It was concluded that oxymetholone kinetics was saturated at 30 mg/kg in female but not male rats. Pathology Findings: A wide spectrum of neoplasms and nonneoplastic lesions was seen in rats administered oxymetholone for 2 years. The incidences of hepatocellular adenoma and hepatocellular adenoma or carcinoma (combined) were significantly increased in 100 mg/kg females as were the incidences of basophilic and clear cell foci in 150 mg/kg males and 100 mg/kg females compared to vehicle controls. The incidences of alveolar/bronchiolar adenoma and adenoma or carcinoma (combined) were significantly increased in 30 mg/kg females. The incidences of mineralization in the lung of 150 mg/kg males and 30 and 100 mg/kg females were significantly increased. The incidence of keratoacanthoma was increased in 30 mg/kg females, and the combined incidence of squamous cell papilloma, keratoacanthoma, basal cell adenoma, squamous cell carcinoma, or carcinoma of the sweat gland was significantly increased in 100 mg/kg females. The incidences of subcutaneous tissue fibroma and fibroma or fibrosarcoma (combined) were significantly increased in 3 mg/kg males. At 2 years, the incidences of benign pheochromocytoma and benign or malignant pheochromocytoma (combined) of the adrenal gland in 150 mg/kg males and medullary hyperplasia in 100 mg/kg females were significantly increased. The incidences of cytoplasmic vacuolization of adrenal cortical cells were significantly increased in 30 and 150 mg/kg males at 18 months and 2 years and in 100 mg/kg females beginning at 12 months and in 30 mg/kg females at 2 years. The incidences of renal tubule adenoma in 3 and 150 mg/kg males were slightly increased. An extended evaluation of the kidney was conducted, and additional incidences of renal tubule adenoma were observed in step sections in vehicle control and dosed male rats. The combined single- and step-section incidence of renal tubule adenoma was significantly increased in 3 mg/kg males. The incidences of nephropathy were significantly increased in 30 and 150 mg/kg males at 2 years and in 100 mg/kg females beginning at 3 months. The severities of nephropathy were significantly increased in dosed groups of males at 2 years and in 100 mg/kg females at 18 months and 2 years. The incidences of mineralization of the kidney were significantly increased in 150 mg/kg males at all time points. The incidences of ovarian dysgenesis were significantly increased in 100 mg/kg females beginning at 3 months and in 30 mg/kg females beginning at 6 months, and severities increased with increasing dose. The incidences of chronic myocardial degeneration (cardiomyopathy) were significantly increased in 100 mg/kg females at 6 months and 2 years and the severity was increased at 2 years. The incidences of lobular hyperplasia were increased in 150 mg/kg males at 18 months and 2 years and in 30 and 100 mg/kg females at all time points. The incidences of seminiferous tubule degeneration were significantly increased in 30 and 150 mg/kg males at 2 years, and the incidences of mineralization of the testis were increased in 150 mg/kg males at 12 months and in 30 mg/kg males at 18 months and at 2 years. Decreased incidences of neoplasms occurred in male and female rats. The incidence of uterine stromal polyp or stromal sarcoma (combined) was significantly decreased in 100 mg/kg females at 2 years. The incidences of mammary gland fibroadenoma and fibroadenoma or carcinoma (combined) were significantly decreased in all dosed groups of females. The incidences of pituitary gland pars distalis adenoma were significantly decreased in 30 and 100 mg/kg females at 2 years. The incidences of testicular interstitial cell adenoma were significantly decreased in 30 and 150 mg/kg males at 18 months and in all dosed groups at 12 months and 2 years. The incidences of mononuclear cell leukemia were significantly decreased in 30 and 150 mg/kg males and 100 mg/kg females at 2 years. GENETIC TOXICOLOGY: Oxymetholone was not mutagenic in S. typhimurium strain TA97, TA98, TA100, or TA1535, with or without S9 metabolic activation. It did not induce chromosomal aberrations in cultured Chinese hamster ovary cells, with or without S9, and no increase in the frequency of micronucleated normochromatic erythrocytes was noted in peripheral blood samples from male or female mice treated for 14 weeks with oxymetholone. CONCLUSIONS: Under the conditions of this 2-year gavage study, there was equivocal evidence of carcinogenic activity of oxymetholone in male F344/N rats based on increased incidences of subcutaneous tissue fibromas and fibromas or fibrosarcomas (combined) of the skin, variably increased incidences of benign and benign or malignant pheochromocytomas (combined) of the adrenal gland, and increased incidences of renal tubule adenomas. There was clear evidence of carcinogenic activity of oxymetholone in female F344/N rats based on increased incidences of hepatocellular neoplasms. Increased incidences of alveolar/bronchiolar neoplasms and skin neoplasms in female rats were also related to oxymetholone administration. Decreased incidences of alveolar/bronchiolar neoplasms and testicular interstitial cell adenomas in males; uterine stromal polyps or stromal sarcomas (combined), mammary gland neoplasms, and pituitary gland pars distalis adenomas in females; and mononuclear cell leukemia in males and females were related to oxymetholone administration. In addition, gavage administration of oxymetholone to male and female F344/N rats resulted in a spectrum of nonneoplastic effects frequently reported with administration of synthetic anabolic androgens. Synonyms: Adroidin; anadroyd; anasteron; anasteronal; anasterone; androstan-3-one, androstano[2,3-c]1,2,5-oxadiazol-17-ol, 17-methyl-, (5-a,17-b)-; becorel; 4,5-dihydro-2-hydroxymethylene-17-a-methyltestosterone; dynasten; HMD; 17b-hydroxy-2- (hydroxymethyl)-17-methyl-5-a-androstan-3-one; 17 hydroxy-2-(hydroxymethylene)-17-methyl-(5-a,17-b)-; 17-hydroxy- 2 (hydroxymethylene)-17-methyl-5-a-17-b-androst-3-one; 17b-hydroxy-2 (hydroxymethylene)-17-a-methyl-5-a-androstan-3-one; 17b-hydroxy-2 (hydroxymethylene)-17-methyl-5a-androstan-3-one; 17-hydroxy-2-(hydroxymethylene) 17-methyl-5-a-17- b-androstan-3-one; 17b-hydroxy-2-hydroxymethylene-17a-methyl-3 androstanone; 2-hydroxymethylene-17-a-methyl-5- a-androstan-17-b-ol-3-one; 2 hydroxymethylene-17a-methyl dihydrotestosterone; 2-hydroxymethylene-17-a-methyl 17-b- hydroxy-3-androstanone; methabol; 17a-methyl-2-hydroxymethylene-17-hydroxy 5-a-androstan-3-one; oximetholonum; oximetolona; oxitosona-50; oxymethenolone; roboral; zenalosyn Trade names: Adroyd; Anadrol; Anapolon; Anapolon 50; Nastenon; Pardroyd; Pavisoid; Plenastril; Protanabol; Synasteron PMID- 12571679 TI - NTP Toxicology and Carcinogenesis Studies 2-Butoxyethanol (CAS NO. 111-76-2) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - 2-Butoxyethanol is a member of a family of ethylene glycol monoalkyl ethers. It is used extensively as a solvent in surface coatings such as lacquers, enamels, varnishes, and latex paint; in paint thinners, paint stripping formulations, and inks; and in degreasers and industrial and household cleaners. 2-Butoxyethanol was nominated for study because of its widespread use in industrial and consumer applications, the potential for exposure to workers and the general population, and the absence of chronic toxicity data. Male and female F344/N rats and B6C3F1 mice were exposed to 2-butoxyethanol (greater than 99% pure) by inhalation (primary route of human exposure) for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and the bone marrow of male F344/N rats and B6C3F1 mice. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to 2-butoxyethanol by inhalation at concentrations of 0, 31, 62.5, 125, 250, or 500 ppm, 6 hours per day, 5 days per week for 14 weeks. One female rat in the 250 ppm group was killed moribund during week 8; four females in the 500 ppm group were killed moribund during week 1 and one during week 5. Final mean body weights of females exposed to 500 ppm were significantly less than those of the chamber controls. Clinical findings included abnormal breathing, pallor, red urine stains, nasal and eye discharge, lethargy, and increased salivation and/or lacrimation. Due to vascular thrombosis and infarction in the tail vertebrae of 500 ppm female rats, the tails became necrotic and either sloughed off or were chewed off. The primary effect on the hematopoietic system was an anemia characterized as macrocytic, normochromic, and regenerative in males exposed to 125 ppm or greater and, to a greater extent, in all exposed groups of females. Compared to the chamber controls, kidney weights of males exposed to 500 ppm and females exposed to 125 ppm or greater and liver weights of males exposed to 250 or 500 ppm and females exposed to 125 ppm or greater were significantly increased, and thymus weights of females exposed to 500 ppm were significantly less. In female rats killed moribund, there was considerable histologic evidence of thrombosis in tissues and organs including the nasal cavity, incisors, liver, lung, and heart. In addition to thrombosis, infarction occurred in the vertebrae of the tail resulting in necrosis and loss of the distal portion of the tail. There were also inflammation, necrosis, and ulceration of the forestomach; necrosis and centrilobular degeneration of the liver; renal tubule degeneration; and atrophy of the spleen and thymus. Exposure related increases in the incidences of Kupffer cell pigmentation, forestomach inflammation and epithelial hyperplasia, bone marrow hyperplasia, splenic hematopoietic cell proliferation, and renal tubule pigmentation were observed in male and/or female rats surviving to the end of the study. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were exposed to 2-butoxyethanol by inhalation at concentrations of 0, 31, 62.5, 125, 250, or 500 ppm, 6 hours per day, 5 days per week for 14 weeks. Two male and two female mice exposed to 500 ppm died and two males and two females were killed moribund during the first 2 weeks of the study. Final mean body weights of 125, 250, and 500 ppm male mice were significantly less than those of the chamber controls. Clinical findings were observed only in 500 ppm males and females that died or were killed moribund and included abnormal breathing, red urine stains, and lethargy. Hematologic evaluation indicated an anemia that was characterized as normocytic, normochromic, and regenerative in mice exposed to 62.5 ppm or greater; the anemia was more pronounced in females. Liver weights of males exposed to 500 ppm were significantly greater than the chamber controls. In mice either dying early or killed moribund, there were inflammation, necrosis, and ulceration of the forestomach; mediastinal pleura and peritoneal inflammationmmation associated with the forestomach lesions; liver necrosis; renal tubule degeneration; atrophy of the spleen, thymus, and mandibular and mesenteric lymph nodes; and degeneration of the testis. Exposure-related increases in the incidences of hematopoietic cell proliferation and hemosiderin pigmentation of the spleen, Kupffer cell hemosiderin pigmentation of the liver, inflammation and epithelial hyperplasia of the forestomach, and renal tubule hemosiderin pigmentation occurred in male and/or female mice surviving to the end of the study. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed to 2 butoxyethanol by inhalation at concentrations of 0, 31.2, 62.5, or 125 ppm, 6 hours per day, 5 days per week for 104 weeks. For hematology and bone marrow analyses, additional groups of 27 male and 27 female rats were exposed to 0, 62.5, or 125 ppm for evaluation at 3, 6, and 12 months and nine male and nine female rats were exposed to 31.2 ppm for evaluation at 3 (hematology only) and 6 months. Survival and Body Weights: Survival of exposed male and female rats was similar to the chamber control groups. The mean body weights of females exposed to 125 ppm were generally less than the chamber control group. Hematology and Bone Marrow Cellularity: The most consistent exposure-related effect on the hematopoietic system was an exposure concentration-related mild macrocytic, normochromic, regenerative anemia present at 3, 6, and 12 months, with females more affected than males. Significant increases in bone marrow cellularity and decreases in the myeloid/erythroid ratio relative to the chamber controls were observed at all time points in females exposed to 125 ppm, and a decrease in the myeloid/erythroid ratio was observed in males exposed to 125 ppm at 12 months. Pathology Findings: The incidence of benign or malignant pheochromocytoma (combined) of the adrenal medulla in females exposed to 125 ppm was not significantly increased compared to the chamber controls but exceeded the historical control range. Exposure-related increases in the incidences of hyaline degeneration of the olfactory epithelium and Kupffer cell pigmentation of the liver were observed in male and female rats. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed to 2-butoxyethanol by inhalation at concentrations of 0, 62.5, 125, or 250 ppm, 6 hours per day, 5 days per week for 104 weeks. For hematology and bone marrow analyses, additional groups of 30 male and 30 female mice were exposed to 0, 62.5, 125, or 250 ppm for evaluation at 3, 6, and 12 months. Survival and Body Weights: Survival of male mice exposed to 125 or 250 ppm was significantly less than that of the chamber control group. The mean body weights of exposed males were generally less than those of the chamber control group during the last 6 months of the study. The mean body weights of exposed female mice were less than those of the chamber control group; the reductions were greater and occurred earlier than those observed in males. Hematology: The most consistent exposure-related effect on the hematopoietic system was an exposure concentration-related minimal normocytic, normochromic, regenerative anemia present at 3, 6, and 12 months, with females affected slightly more than males. Pathology Findings: In females exposed to 250 ppm, incidences of forestomach squamous cell papilloma and squamous cell papilloma or carcinoma (combined) were significantly increased relative to the chamber controls, and these incidences exceeded the ranges in historical chamber controls. In 2-butoxyethanol exposed males, there were possible exposure-related increases in the incidences of squamous cell papilloma of the forestomach, although the increases were not significant and the incidences were within the historical control range for chamber controls. Accompanying these neoplasms in females and, to a lesser extent, in males were exposure-related increases in the incidences of ulcer and epithelial hyperplasia of the forestomach. In male mice exposed to 250 ppm, the incidence of hemangiosarcoma of the liver was significantly increased relative to chamber controls and exceeded the range in historical controls; in addition, there were possible exposure-related increases in the incidence of hepatocellular carcinoma. Incidences of hemosiderin pigmentation in the Kupffer cells were significantly increased in 125 and 250 ppm males and all exposed groups of females. The incidences of splenic hematopoietic cell proliferation and hemosiderin pigmentation were generally increased in males and females, and the incidences of bone marrow hyperplasia were increased in males. The incidences of hyaline degeneration of the olfactory and respiratory epithelia of the nose were increased in female mice. GENETIC TOXICOLOGY: 2 Butoxyethanol did not induce mutations in any of the S. typhimurium strains tested, with or without induced hamster or rat liver S9. 2-Butoxyethanol induced cycle delay but did not induce either sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells with or without S9. 2 Butoxyethanol did not induce micronuclei in bone marrow cells of male rats or mice administered the chemical by intraperitoneal injection three times at 24 hour intervals. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was no evidence of carcinogenic activity of 2-butoxyethanol in male F344/N rats exposed to 31.2, 62.5, or 125 ppm. There was equivocal evidence of carcinogenic activity of 2-butoxyethanol in female F344/N rats based on the increased combined incidences of benign or malignant pheochromocytoma (mainly benign) of the adrenal medulla. There was some evidence of carcinogenic activity of 2-butoxyethanol in male B6C3F1 mice based on increased incidences of hemangiosarcoma of the liver. A marginal increase in the incidences of forestomach squamous cell papilloma and an increase in the incidences of hepatocellular carcinoma may have been exposure related. There was some evidence of carcinogenic activity of 2-butoxyethanol in female B6C3F1 mice based on increased incidences of fore stomach squamous cell papilloma or carcinoma (mainly papilloma). Increased incidences of forestomach neoplasms in male and female mice occurred in groups in which ulceration and hyperplasia were also present. Exposure to 2-butoxyethanol caused a mild regenerative anemia and effects secondary to the anemia. Synonyms: 2-Butoxy-1-ethanol; m-butyl ether; butyl glycol; ethylene glycol monobutyl ether Trade name: Butyl Cellosolve PMID- 12571680 TI - NTP Toxicology and Carcinogenesis Studies of Pentachlorophenol (CAS NO. 87-86-5) in F344/N Rats (Feed Studies). AB - Pentachlorophenol has been used as an herbicide, algicide, defoliant, wood preservative, germicide, fungicide, and molluscicide. Pentachlorophenol was nominated by the National Cancer Institute for carcinogenicity testing based on its widespread use as a wood preservative, potential for entering the environment (pentachlorophenol residues have been found worldwide in soil, water, and air samples; in food products; and in human and animal tissues and body fluids), and likelihood of bioaccumulation in the environment (pentachlorophenol is persistent in soil, having a half-life of up to 5 years). Technical Report No. 349 contains the results of the 2-year studies of pentachlorophenol performed by the NTP with B6C3F1 mice. Male and female F344/N rats were exposed to pentachlorophenol (approximately 99% pure) in feed for 28 days or 2 years. Genetic toxicology studies were conducted in vitro in Salmonella typhimurium and cultured Chinese hamster ovary cells and in vivo in rat and mouse bone marrow cells. 28-DAY STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were given 0, 200, 400, 800, 1,600, or 3,200 ppm pentachlorophenol, equivalent to average daily doses of approximately 20, 40, 75, 150, or 270 mg pentachlorophenol/kg body weight to males and females in feed for 28 days. With the exception of one male and two females exposed to 3,200 ppm, all rats survived until the end of the study. The final mean body weights and body weight gains of male rats exposed to 1,600 or 3,200 ppm and female rats exposed to 400, 800, 1,600, or 3,200 ppm were significantly less than those of the controls; rats exposed to 3,200 ppm lost weight during the study. Feed consumption by 3,200 ppm males was less than that by the control group throughout the study. The absolute and relative liver weights of 400, 800, and 1,600 ppm males and all exposed groups of females were significantly greater than those of the controls. Compared to the control groups, the incidences of minimal to mild hepatocyte degeneration in males and females exposed to 400 ppm or greater and the incidences of centrilobular hepatocyte hypertrophy in the 3,200 ppm groups were increased. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were fed diets containing 200, 400, or 600 ppm pentachlorophenol (equivalent to average daily doses of approximately 10, 20, and 30 mg/kg) for 105 weeks. Stop-exposure groups of 60 male and 60 female rats received 1,000 ppm (equivalent to 60 mg/kg) in feed for 52 weeks, after which animals received undosed feed for the remainder of the 2-year study; 10 male and 10 female control and 1,000 ppm rats were evaluated at 7 months. Survival, Body Weights,and Feed Consumption: In the 2-year study, survival of 600 and 1,000 ppm males was greater than that of the controls. Mean body weights of 400 and 600 ppm males and females were generally less than those of controls. When exposure to pentachlorophenol was discontinued at week 52, mean body weights of 1,000 ppm males and females were 17%% and 22%% lower than those of the respective controls; however, by the end of week 87, the mean body weights were similar to those of the controls. Generally, feed consumption by exposed groups was similar to that by the controls. Pathology Findings: At 2 years, the incidence of malignant mesothelioma originating from the tunica vaginalis was significantly greater in 1,000 ppm males than in the controls, and the incidence exceeded the historical control range. Nasal squamous cell carcinomas were present in one control male, three 200 ppm males, one 400 ppm male, and five 1,000 ppm males at 2 years, and the incidence in 1,000 ppm males exceeded the historical control range. At the 7 month interim evaluation, the incidences of centrilobular hepatocyte hypertrophy in 1,000 ppm males and females and hepatocyte cytoplasmic vacuolization in 1,000 ppm males was significantly greater than those in the controls. At 2 years, the incidences of several nonneoplastic liver lesions including hepatodiaphragmatic nodules and hepatocyte cystic degeneration in all exposed ation in all exposed groups of males and basophilic foci in 1,000 ppm males were increased compared to the controls. GENETIC TOXICOLOGY: Pentachlorophenol (91.6%% pure) was tested in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 at doses up to 30 μg/plate with and without induced rat or hamster liver S9; no significant increases in the number of revertant colonies were observed in any of the strain/activation combinations. When tested for cytogenetic effects in cultured Chinese hamster ovary cells, pentachlorophenol was weakly positive for induction of sister chromatid exchanges and chromosomal aberrations. In the sister chromatid exchange test, a weakly positive response was observed within a concentration range of 3 to 30 μg/mL in the absence of S9; with S9, no induction of sister chromatid exchanges was noted. In the chromosomal aberrations test, pentachlorophenol was negative without S9 but induced small but significant increases in the frequency of aberrant cells in the presence of S9 at doses of 80 and 100 μg/mL. In contrast to the positive in vitro results in the test for induction of chromosomal aberrations, no increase in the frequency of micronucleated erythrocytes was noted in bone marrow of male rats or mice administered pentachlorophenol by intraperitoneal injection three times at 24 hour intervals. The highest dose administered to rats (75 mg/kg) and mice (150 mg/kg) was lethal. CONCLUSIONS: Under the conditions of this 2-year feed study, there was no evidence of carcinogenic activity of pentachlorophenol in male or female F344/N rats fed diets containing 200, 400, or 600 ppm. There was some evidence of carcinogenic activity of pentachlorophenol in male F344/N rats given feed containing 1,000 ppm for 1 year followed by control feed for 1 year (stop exposure study), based on increased incidences of mesothelioma and nasal squamous cell carcinoma. There was no evidence of carcinogenic activity of pentachlorophenol in female rats given feed containing 1,000 ppm for 1 year and maintained on control feed for 1 year. Stop-exposure males and females recovered from a transitory reduction in body weight gain by the end of the 2-year study, and males had increased survival compared to the controls. Synonyms: Chlorophen; PCP; penchlorol; penta; pentachlorofenol; pentachlorofenolo; 2,3,4,5,6 pentachlorophenol Trade names: Acutox; Chem-Penta; Chem-Tol; Cryptogil ol; Dowicide 7; Dowicide EC-7; Dow Pentachlorophenol DP-2 Antimicrobial; Durotox; EP 30; Fungifen; Fungol; Glazd Penta; Grundier Arbezol Lauxtol; Lauxtol A; Liroprem; Moosuran; Pentacon; Penta-Kil; Pentasol; Penwar; Peratox; Permacide; Permagard; Permasan; Permatox; Priltox; Permite; Santophen; Santophen 20; Sinituho; Term-i Trol; Thompson's Wood Fix; Weedone; Witophen P PMID- 12571681 TI - Toxicology and Carcinogenesis Studies of Furfuryl Alcohol (CAS No. 98-00-0) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Furfuryl alcohol-based resins are used as binding agents in foundry sand and as corrosion inhibitors in mortar, grout, and cement. Because of their heat resistance, furan resins are used in the manufacture of fiberglass-reinforced plastic equipment. Furfuryl alcohol was selected for evaluation because of the absence of data on its carcinogenic potential and its large production volume, widespread use in manufacturing, and ubiquitous presence in consumer goods. Male and female F344/N rats and B6C3F1 mice were exposed to furfuryl alcohol (greater than 98% pure) by inhalation for 16 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and mouse bone marrow cells. 16-DAY STUDY IN RATS: Groups of five male and five female rats were exposed to concentrations of 0, 16, 31, 63, 125, or 250 ppm furfuryl alcohol by inhalation, 6 hours per day, 5 days per week for 16 days. All male and female rats exposed to 250 ppm died by day 2 of the study, and one male rat exposed to 125 ppm died on day 5. Final mean body weights of male and female rats exposed to 125 ppm were significantly less than those of the chamber control groups. Male rats exposed to 31, 63, or 125 ppm and female rats exposed to 125 ppm gained less weight than the chamber control groups. Clinical findings included dyspnea, hypoactivity, and nasal and ocular discharge in males and females exposed to 63, 125, or 250 ppm. All exposed animals developed lesions in the nasal respiratory epithelium and olfactory epithelium, and the severities of these lesions generally increased with increasing exposure concentration. 16-DAY STUDY IN MICE: Groups of five male and five female mice were exposed to concentrations of 0, 16, 31, 63, 125, or 250 ppm furfuryl alcohol by inhalation, 6 hours per day, 5 days per week for 16 days. All male and female mice exposed to 250 ppm died by day 4 of the study, and one female mouse exposed to 125 ppm died on day 14. Mean body weights of male and female mice exposed to 63 or 125 ppm were significantly less than those of the chamber control groups. All exposed animals except one 16 ppm male developed lesions in the nasal respiratory epithelium and/or olfactory epithelium, and the severities of these lesions generally increased with increasing exposure concentration. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to furfuryl alcohol at concentrations of 0, 2, 4, 8, 16, or 32 ppm, 6 hours per day, 5 days per week for 14 weeks. All rats survived to the end of the study. The mean body weight gain of females exposed to 32 ppm was less than that of the chamber control group. Exposure-related increases in the incidences of squamous metaplasia of the respiratory and transitional epithelium, goblet cell hyperplasia of the respiratory epithelium, and hypertrophy of the respiratory epithelium lining the nasopharyngeal duct were observed in the nose of male and female rats. The incidences of degeneration, hyperplasia, metaplasia, and surface exudate of the olfactory epithelium generally increased with increasing exposure concentration in males and females. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were exposed to furfuryl alcohol at concentrations of 0, 2, 4, 8, 16, or 32 ppm, 6 hours per day, 5 days per week for 14 weeks. All mice survived to the end of the study. Heart weights of 32 ppm males were significantly less than those of the chamber controls. Exposure-related histologic changes included degeneration, metaplasia, and chronic inflammation of the olfactory epithelium; hyaline droplets of the respiratory epithelium; and squamous metaplasia of the submucosal gland of the cuboidal epithelium in males and females. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed to furfuryl alcohol by inhalation, 6 hours per day, 5 days per week for 105 weeks, at concentrations of 0, 2, 8, or 32 ppm. Survival and Body Weights All male rats exposed to 32 ppm died by week 99; survival of all other exposed groups of male and femald female rats was similar to that of the chamber control groups. Mean body weights of 32 ppm males were less than those of the chamber control group beginning at week 19. Pathology Findings All groups of exposed male and female rats had significantly increased incidences of nonneoplastic histologic changes of the nose compared to the chamber control groups. An adenoma of the lateral wall of the nose was observed in one 2 ppm male and one 8 ppm female, an adenoma of the respiratory epithelium was observed in one 8 ppm male and one 32 ppm female, one carcinoma of the respiratory epithelium was observed in a 32 ppm male, and squamous cell carcinomas of the nose were observed in three 32 ppm males. Renal tubule adenomas were present in one chamber control male, one 2 ppm male, two 8 ppm males, and two 32 ppm females. One 2 ppm female had a renal tubule carcinoma. Additional histologic sections from the kidney revealed the presence of additional hyperplasias in all groups of males and females; one additional renal tubule adenoma was observed in each of the chamber control, 2 ppm, and 8 ppm male groups, and four additional adenomas were observed in 32 ppm males. In females, two additional adenomas were found in the 8 ppm group, one adenoma in the 32 ppm group, and one carcinoma in the 2 ppm group. The severities of nephropathy relative to the chamber controls were increased in 32 ppm males and females. Males exposed to 32 ppm had extrarenal signs indicative of marked nephropathy including parathyroid gland hyperplasia and fibrous osteodystrophy. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed to furfuryl alcohol by inhalation, 6 hours per day, 5 days per week for 105 weeks, at concentrations of 0, 2, 8, or 32 ppm. Survival, Body Weights, and Clinical Findings Survival of exposed males and females was similar to that of the chamber control groups. Mean body weights of exposed males were generally similar to those of the chamber control group throughout the study. Mean body weights of exposed females were less than those of the chamber control group during year 2 of the study. Female mice exposed to 32 ppm developed focal corneal opacities. Pathology Findings The incidences of renal tubule neoplasms were increased in 32 ppm male mice compared to the chamber control group and exceeded the historical control range for inhalation studies. Step sectioning revealed the presence of additional hyperplasias in the chamber control and exposed groups and one adenoma in 32 ppm males. The severity of nephropathy increased with increasing exposure concentration in male mice. The incidence of renal tubule degeneration in male mice exposed to 32 ppm was significantly greater than in the chamber control group. Incidences of a variety of nonneoplastic lesions of the nose were significantly greater in all exposed groups of male and female mice than in the chamber control groups. The incidence of degeneration of the cornea was significantly greater in 32 ppm female mice compared to the chamber control group. GENETIC TOXICOLOGY: Furfuryl alcohol was not mutagenic in Salmonella typhimurium strain TA98, TA100, TA1535, or TA1537, with or without S9. It did induce sister chromatid exchanges in cultured Chinese hamster ovary cells in the absence of S9, but not in the presence of S9. No induction of chromosomal aberrations was noted in cultured Chinese hamster ovary cells treated with furfuryl alcohol in the absence of S9, but in the presence of S9 an equivocal result was obtained. In vivo, no induction of sister chromatid exchanges, chromosomal aberrations, or micronuclei was noted in bone marrow cells of male mice after treatment with furfuryl alcohol. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was some evidence of carcinogenic activity of furfuryl alcohol in male F344/N rats based on increased incidences of combined neoplasms of the nose. There was equivocal evidence of carcinogenic activity of furfuryl alcohol in female F344/N rats based on marginally increased incidences of neoplasms of the nose and renal tubule neoplasms. There was some evidence of carcinogenic activity of furfuryl alcohol in male B6C3F1 mice based on increased inci dences of renal tubule neoplasms. There was no evidence of carcinogenic activity of furfuryl alcohol in female B6C3F1 mice exposed to 2, 8, or 32 ppm. Exposure of male and female rats and male mice to furfuryl alcohol was associated with increased incidences of nonneoplastic lesions of the nose and increased severities of nephropathy. Exposure of female mice to furfuryl alcohol was associated with increased incidences of nonneoplastic lesions of the nose and corneal degeneration. Synonyms: 2-Furancarbinol; 2-furanmethanol, furfuralcohol, a-furylcarbinol; 2-hydroxymethylfuran PMID- 12571682 TI - NTP Toxicology and Carcinogenesis Studies of Oleic Acid Diethanolamine Condensate (CAS No. 93-83-4) in F344/N Rats and B6C3F1 Mice (Dermal Studies). AB - Oleic acid diethanolamine condensate is widely used as an emollient, thickener, and foam stabilizer present in cosmetic formulations of bath additives, shampoos, conditioners, lipsticks, and hair dyes. Male and female F344/N rats and B6C3F1 mice received dermal applications of diethanolamine in 95% ethanol for 13 weeks or 2 years. Genetic toxicology studies were performed in Salmonella typhimurium and L5178Y mouse lymphoma cells. 13-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were admin istered 0, 25, 50, 100, 200, or 400 mg oleic acid diethanolamine condensate/kg body weight in ethanol dermally for 13 weeks. All male and female rats survived until the end of the study. The final mean body weights and body weight gains of 200 and 400 mg/kg males and the mean body weight gain of 400 mg/kg females were significantly less than those of the vehicle controls. The only chemical-related clinical finding was irritation of the skin at the site of application in most males administered 100 mg/kg or greater and in all females administered 50 mg/kg or greater. Segmented neutrophil counts were increased relative to the vehicle controls in the 400 mg/kg male group on days 5 and 19, in the 200 mg/kg female group on day 19 and at week 13, and in the 400 mg/kg female group on days 5 and 19 and at week 13. Alkaline phosphatase concentrations were significantly increased in the 200 mg/kg male group on day 19, the 200 mg/kg female group at week 13, and in the 400 mg/kg groups of males and females at week 13. Kidney weights of 200 and 400 mg/kg females were significantly greater than those of the vehicle controls. Lesions of the skin at the site of application included epidermal hyperplasia, parakeratosis, chronic active dermal inflammation, suppurative epidermal inflammation, and sebaceous gland hypertrophy in dosed rats. The severities of these lesions generally increased with increasing dose. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were admin istered 0, 50, 100, 200, 400, or 800 mg oleic acid diethanolamine condensate/kg body weight in ethanol dermally for 13 weeks. All male and female mice except one 800 mg/kg male survived until the end of the study. Final mean body weights and body weight gains of 800 mg/kg males and females and 400 mg/kg females were significantly less than those of the vehicle controls. Clinical findings in dosed mice included irritation of the skin at the site of application. Irritation occurred in all surviving dosed males and in most females administered 100 mg/kg or greater and progressed to ulcer in one 800 mg/kg male. The heart weights of 400 and 800 mg/kg males and females and 200 mg/kg females and the kidney weights of 50, 100, and 400 mg/kg males were significantly greater than those of the vehicle controls. Relative to the vehicle controls, the liver weights were increased in all dosed groups. Lesions of the skin at the site of application in dosed mice included epidermal hyperplasia, parakeratosis, suppurative epidermal inflammation, chronic active dermal inflammation, sebaceous gland hypertrophy, and ulcer. The severities of these lesions generally increased with increasing dose. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were administered 0, 50, or 100 mg oleic acid diethanolamine condensate/kg body weight in ethanol dermally for 2 years. Survival, Body Weights, and Clinical Findings: Survival of dosed male and female rats was similar to that of the vehicle control groups. Mean body weights of 100 mg/kg males were slightly less than those of the vehicle controls throughout most of the study. Mean body weights of 100 mg/kg females were less than those of the vehicle controls beginning at week 24. The only significant treatment-related clinical finding was mild to moderate irritation of the skin at the site of application in dosed males and females. Pathology Findings: The predominant effects of oleic acid diethanolamine condensate administration were minimal to moderate nonneoplastic lesions of the skin at the site of application in dosed rats. These lesioe lesions included epidermal hyperplasia, sebaceous gland hyperplasia, hyperkeratosis, parakeratosis, chronic active dermal inflammation, and ulcer. 2-YEAR STUDY IN MICE: Groups of 55 male and 55 female mice were administered 0, 15, or 30 mg oleic acid diethanolamine condensate/kg body weight in ethanol dermally for 2 years. Five animals from each group were evaluated at 3 months for gross lesions and skin histopathology. Survival, Body Weights, and Clinical Findings: Survival of dosed male and female mice was similar to that of the vehicle control groups. Mean body weights of dosed males and of 15 mg/kg females were similar to those of the vehicle controls throughout the study. Mean body weights of 30 mg/kg females were less than those of the vehicle controls from week 76 until the end of the study. The only significant treatment-related clinical finding was irritation of the skin at the site of application in 30 mg/kg males. Pathology Findings: The incidences of epidermal hyperplasia, sebaceous gland hyperplasia, and chronic active inflammation of the dermis in all dosed groups were significantly increased relative to the vehicle controls at 3 months and at 2 years. The increased incidences of hyperkeratosis in dosed males at 3 months and in dosed males and females at 2 years, of parakeratosis in 30 mg/kg males at 3 months and 2 years, and of ulcer in 30 mg/kg males and exudate in 30 mg/kg males and females at 2 years were also attributed to chem ical administration. GENETIC TOXICOLOGY: Oleic acid diethanolamine condensate was not mutagenic in S. typhimurium strain TA97, TA98, TA100, or TA1535, with or without S9 metabolic activation enzymes. In addition, it did not induce mutations in mouse L5178Y lymphoma cells treated with or without S9. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of oleic acid diethanolamine condensate in male or female F344/N rats administered 50 or 100 mg/kg or in male or female B6C3F1 mice administered 15 or 30 mg/kg. Dermal administration of oleic acid diethanolamine condensate to male and female rats was associated with epidermal hyperplasia, sebaceous gland hyper plasia, hyperkeratosis, parakeratosis, chronic active inflammation of the dermis, and ulceration of the skin at the site of application. Dermal administration of oleic acid diethanolamine condensate to mice was associated with epidermal hyperplasia, sebaceous gland hyperplasia, hyperkeratosis, chronic active inflammation of the dermis, and exudate of the skin at the site of application in males and females and parakeratosis and ulcer of the skin at the site of application in males. Synonyms: Diethanolamine oleate; diethanolammonium oleate; (Z)-9-octadecenoic acid, compound with 2,2.-imnobis(ethanol) (1:1); oleamide diethanolamine PMID- 12571683 TI - NTP Toxicology and Carcinogenesis Studies of Lauric Acid Diethanolamine Condensate (CAS NO. 120-40-1) in F344/N Rats and B6C3F1 Mice (Dermal Studies). AB - Lauric acid diethanolamine condensate is widely used in cosmetics, shampoos, soaps, and related consumer products, to which there is extensive human exposure. Because of the lack of information about potential risks associated with long term exposure, lauric acid diethanolamine condensate, coconut oil acid diethanolamine condensate, and oleic acid diethanolamine condensate were selected as representative of the class of diethanolamides for evaluation of prechronic toxicity and carcinogenic potential. Male and female F344/N rats and B6C3F1 mice were exposed to lauric acid diethanolamine condensate dermally for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, L5178Y mouse lymphoma cells, cultured Chinese hamster ovary cells, and mouse peripheral blood erythrocytes. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were admin istered 0, 25, 50, 100, 200, or 400 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 14 weeks. All animals survived until study termination. Final mean body weights and body weight gains of males receiving 200 or 400 mg/kg were significantly less than those of the vehicle control group. Irritation of the skin at the site of application was observed in males receiving 100 mg/kg or greater and in females receiving 200 or 400 mg/kg. Kidney weights of females administered 200 or 400 mg/kg were significantly greater than those of the vehicle control group. There were dose-dependent increases in the incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, chronic inflammation, parakeratosis, and ulcer. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were admin istered 0, 50, 100, 200, 400, or 800 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 14 weeks. All animals survived until the end of the study, and final mean body weights and body weight gains of dosed mice were generally similar to those of the vehicle control groups. Irritation of the skin at the site of application was observed in all males and females administered 400 or 800 mg/kg. The kidney weights of males receiving 100, 400, or 800 mg/kg and females receiving 800 mg/kg were significantly greater than those of the vehicle controls. Liver weights of females administered 200 mg/kg or greater were significantly greater than those of vehicle controls. Increased incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, chronic inflammation, parakeratosis, and ulcer, were observed in males and females receiving 200 mg/kg or greater. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were admin istered 0, 50, or 100 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 104 or 105 weeks. Survival and Body Weights There were no significant differences between vehicle control and dosed males or females in survival or mean body weights. Pathology Findings There were no chemical-related differences in neoplasm incidences. Dose-related increases occurred in the incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, hyperkeratosis, chronic inflammation, parakeratosis, and ulcer. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were admin istered 0, 100, or 200 mg lauric acid diethanolamine condensate/kg body weight in ethanol by dermal application for 105 or 106 weeks. Survival and Body Weights There were no significant differences in survival between vehicle control and dosed males or females. Mean body weights of females that received 200 mg/kg were less than those of the vehicle controls beginning at week 33. Pathology Findings The incidences of hepatocellular adenoma or carcinoma (combined) were significantly increased in dosed females compared to the vehicle controls, as was the incidence of hepatocellular adenoma in the 100 mg/kg female group. There were dose-related increases in the incidences of nonneoplastic lesions of the skin at the site of application, including epidermal and sebaceous gland hyperplasia, hyperkeratosis, chronic inflammation, and parakeratosis. Dosed males had greater incidences of thyroid gland follicular cell focal hyperplasia than did the vehicle controls. GENETIC TOXICOLOGY: Lauric acid diethanolamine condensate was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100, or TA1535, with or without S9 metabolic activation enzymes. No increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was noted after exposure to lauric acid diethanolamine condensate, with or without S9. In cytogenetic tests with cultured Chinese hamster ovary cells, lauric acid diethanolamine condensate was shown to induce sister chromatid exchanges, but not chromosomal aberrations, with and without S9. In vivo, no increase in the frequency of micro nucleated normochromatic erythrocytes was observed in peripheral blood samples from male and female mice treated dermally with lauric acid diethanolamine condensate for 14 weeks. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of lauric acid diethanolamine condensate in male or female F344/N rats administered 50 or 100 mg/kg or in male B6C3F1 mice administered 100 or 200 mg/kg. There was some evidence of carcinogenic activity in female B6C3F1 mice based on increased incidences of hepatocellular neoplasms. These increases were associated with free diethanolamine, which was present as a contaminant of lauric acid diethanolamine condensate. Dermal administration of lauric acid diethanolamine condensate to rats and mice for 2 years resulted in increased incidences of epidermal and sebaceous gland hyperplasia, hyperkeratosis, chronic inflammation, and parakeratosis at the site of application. Lauric acid diethanolamine condensate administration also resulted in increased incidences of thyroid gland follicular cell hyperplasia in dosed male mice. Synonyms: N,N-bis(2-hydroxyethyl) dodecanamide; N,N-bis(hydroxyethyl) lauramide; N,N-bis(b-hydroxyethyl) lauramide; bis(2-hydroxyethyl) lauramide; coco diethanolamide; coconut oil amide of diethanolamine; diethanollauramide; N,N-diethanollauramide; N,N-diethanollauric acid amide; lauramide DEA; lauric diethanolamide; lauroyl diethanolamide; lauryl diethanolamide; LDA; LDE Trade names: Clindrol 200 L; Ninol AA62; Onyxol 345; Rewomid DLMS; Rewomid DL 203/S; Richamide 6310; Rolamid CD; Standamidd LD; Steinamid DL 203 S; Super amide L-9A; Super amide L-9C; Synotol L-60; Unamide J 56; Varamid ML 1. PMID- 12571684 TI - Toxicology and carcinogenesis studies of coconut oil acid diethanolamine condensate (CAS No. 68603-42-9) in F344/N rats and B6C3F1 mice (dermal studies). AB - Coconut oil acid diethanolamine condensate, a mixture of fatty acid diethanolamides of the acids found in coconut oil, is widely used in cosmetics, shampoos, soaps, and related consumer products. Because of the lack of information about potential risks associated with long-term exposure, coconut oil acid diethanolamine condensate was selected as a representative of the diethanolamine chemical class for evaluation of toxicity and carcinogenic potential. Male and female F344/N rats and B6C3F1 mice received dermal applications of coconut oil acid diethanolamine condensate for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, L5178Y mouse lymphoma cells, cultured Chinese hamster ovary cells, and mouse peripheral blood erythrocytes. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats received dermal applications of 0, 25, 50, 100, 200, or 400 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol, five times per week for 14 weeks. All rats survived until the end of the study. Final mean body weights and body weight gains of 200 and 400 mg/kg males and females were significantly less than those of the vehicle controls. Clinical findings included irritation of the skin at the site of application in 100, 200, and 400 mg/kg males and females. Cholesterol concentrations were significantly decreased in 200 and 400 mg/kg males and in females administered 100 mg/kg or greater; triglyceride concentrations were also decreased in 200 and 400 mg/kg males. Histopathologic lesions of the skin at the site of application included epidermal hyperplasia, sebaceous gland hyperplasia, chronic active inflammation, parakeratosis, and ulcer. The incidences and severities of these skin lesions generally increased with increasing dose in males and females. The incidences of renal tubule regeneration in 100, 200, and 400 mg/kg females were significantly greater than the vehicle control incidence, and the severities in 200 and 400 mg/kg females were increased. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice received dermal applications of 0, 50, 100, 200, 400, or 800 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol, five times per week for 14 weeks. All mice survived until the end of the study. Final mean body weights and body weight gains of dosed males and females were similar to those of the vehicle controls. The only treatment-related clinical finding was irritation of the skin at the site of application in males and females administered 800 mg/kg. Weights of the liver and kidney of 800 mg/kg males and females, the liver of 400 mg/kg females, and the lung of 800 mg/kg females were significantly increased compared to the vehicle controls. Epididymal spermatozoal concentration was significantly increased in 800 mg/kg males. Histopathologic lesions of the skin at the site of application included epidermal hyperplasia, sebaceous gland hyperplasia, chronic active inflammation, parakeratosis, and ulcer. The incidences and severities of these skin lesions generally increased with increasing dose in males and females. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats received dermal applications of 0, 50, or 100 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol five times a week for 104 weeks. Survival, BODY WEIGHTS, AND CLINICAL FINDINGS: The survival rates of treated male and female rats were similar to those of the vehicle controls. The mean body weights of dosed males and females were similar to those of the vehicle controls throughout most of the study. The only chemical-related clinical finding was irritation of the skin at the site of application in 100 mg/kg females. PATHOLOGY FINDINGS: There were marginal increases in the incidences of renal tubule adenoma or carcinoma (combined) in 50 mg/kg females. The severity of nephropathy increased with increasing dose in female rats. Nonneoplastic lesions of the skin at the site of application included epidermal hyperplasia, sebaceous gland hyperplasia, parakeratosis, and hyperkeratosis, and the incidences and severities of these lesions increased with increasing dose. The incidences of chronic active inflammation, epithelial hyperplasia, and epithelial ulcer of the forestomach increased with dose in female rats, and the increases were significant in the 100 mg/kg group. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice received dermal applications of 0, 100, or 200 mg coconut oil acid diethanolamine condensate/kg body weight in ethanol five times a week for 104 to 105 weeks. SURVIVAL, BODY WEIGHTS, AND CLINICAL FINDINGS: Survival of dosed male and female mice was generally similar to that of the vehicle controls. Mean body weights of 100 mg/kg females from week 93 and 200 mg/kg females from week 77 were less than those of the vehicle controls. The only clinical finding attributed to treatment was irritation of the skin at the site of application in males administered 200 mg/kg. PATHOLOGY FINDINGS: The incidences of hepatic neoplasms (hepatocellular adenoma, hepatocellular carcinoma, and hepatoblastoma) were significantly increased in male and/or female mice. Most of the incidences exceeded the historical control ranges. The incidences of eosinophilic foci in dosed groups of male mice were increased relative to that in the vehicle controls. The incidences of renal tubule adenoma and renal tubule adenoma or carcinoma (combined) were significantly increased in 200 mg/kg males. Several nonneoplastic lesions of the skin at the site of application were considered treatment related. Incidences of epidermal hyperplasia, sebaceous gland hyperplasia, and hyperkeratosis were greater in all dosed groups of males and females than in the vehicle controls. The incidences of ulcer in 200 mg/kg males and inflammation and parakeratosis in 200 mg/kg females were greater than those in the vehicle controls. The incidences of thyroid gland follicular cell hyperplasia in all dosed groups of males and females were significantly greater than those in the vehicle control groups. GENETIC TOXICOLOGY: Coconut oil acid diethanolamine condensate did not show genotoxic activity in vitro. It was not mutagenic in Salmonella typhimurium, nor did it produce an increase in mutant L5178Y mouse lymphoma cell colonies. In addition, no increases in the frequencies of sister chromatid exchanges or chromosomal aberrations were observed in Chinese hamster ovary cells after incubation with coconut oil acid diethanolamine condensate. All these in vitro assays were conducted with and without induced S9 activation enzymes. In contrast to the uniformly negative results in vitro, positive results were obtained in a peripheral blood micronucleus test in male and female mice from the 14-week dermal study. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of coconut oil acid diethanolamine condensate in male F344/N rats administered 50 or 100 mg/kg. There was equivocal evidence of carcinogenic activity in female F344/N rats based on a marginal increase in the incidences of renal tubule neoplasms. There was clear evidence of carcinogenic activity in male B6C3F1 mice based on increased incidences of hepatic and renal tubule neoplasms and in female B6C3F1 mice based on increased incidences of hepatic neoplasms. These increases were associated with the concentration of free diethanolamine present as a contaminant in the diethanolamine condensate. Exposure of rats to coconut oil acid diethanolamine condensate by dermal application in ethanol for 2 years resulted in epidermal hyperplasia, sebaceous gland hyperplasia, hyperkeratosis, and parakeratosis in males and females and ulcer in females at the site of application. There were increases in the incidences of chronic inflammation, epithelial hyperplasia, and epithelial ulcer in the forestomach of female rats. The severities of nephropathy in dosed female rats were increased. Exposure of mice to coconut oil acid diethanolamine condensate by dermal application for 2 years resulted in increased incidences of eosinophilic foci of the liver in males. Increased incidences of epidermal hyperplasia, sebaceous gland hyperplasia, and hyperkeratosis in males and females, ulcer in males, and parakeratosis and inflammation in females at the site of application and of follicular cell hyperplasia in the thyroid gland of males and females were chemical related. PMID- 12571685 TI - NTP Toxicology and Carcinogenesis Studies of Diethanolamine (CAS No. 111-42-2) in F344/N Rats and B6C3F1 Mice (Dermal Studies). AB - Diethanolamine is widely used in the preparation of diethanolamides and diethanolamine salts of long-chain fatty acids that are formulated into soaps and surfactants used in liquid laundry and dishwashing detergents, cosmetics, shampoos, and hair conditioners. Diethanolamine is also used in textile processing, in industrial gas purification to remove acid gases, as an anticorrosion agent in metalworking fluids, and in preparations of agricultural chemicals. Aqueous diethanolamine solutions are used as solvents for numerous drugs that are administered intravenously. Diethanolamine was selected for evaluation because its large-scale production and pattern of use indicate the potential for widespread human exposure. Male and female F344/N rats and B6C3F1 mice received dermal applications of diethanolamine in 95% ethanol for 2 years. Genetic toxicology studies were performed in Salmonella typhimurium, L5178Y mouse lymphoma cells, cultured Chinese hamster ovary cells, and B6C3F1 mouse peripheral blood erythrocytes. RATS: Groups of 50 male rats were administered 0, 16, 32, or 64 mg diethanolamine/kg body weight in ethanol dermally for 2 years. Groups of 50 female rats were administered 0, 8, 16, or 32 mg/kg in ethanol dermally for 2 years. Survival, Body Weights, and Clinical Findings Survival of vehicle control and dosed male and female rats was similar. Mean body weights of 64 mg/kg males were less than those of the vehicle controls beginning week 8, and mean body weights of females were generally similar to those of the vehicle control group. The only clinical finding attributed to diethanolamine administration was irritation of the skin at the site of application. Pathology Findings: Minimal to mild nonneoplastic lesions occurred at the site of application in the epidermis of dosed male and female rats. The incidence of acanthosis in 64 mg/kg males, the incidences of hyperkeratosis in 32 and 64 mg/kg males and in all dosed female groups, and the incidences of exudate in 64 mg/kg males and in all dosed female groups were greater than those in the controls. The incidences and severities of nephropathy were significantly increased in dosed female rats compared to the vehicle controls. MICE: Groups of 50 male and 50 female mice were administered 0, 40, 80, or 160 mg diethanolamine/kg body weight in ethanol dermally for 2 years. Survival, Body Weights, and Clinical Findings Survival of dosed male groups was similar to that of the vehicle control group; survival of dosed female groups was significantly less than that of the vehicle control group. Mean body weights of 80 and 160 mg/kg males were less than those of the vehicle controls after weeks 88 and 77, respectively. Mean body weights of dosed groups of females were generally less than those of the vehicle controls during the second year of the study. Pathology Findings: In male mice, the incidences of hepatocellular adenoma and of hepatocellular adenoma or carcinoma (combined) in all dosed groups and of hepatocellular carcinoma and hepatoblastoma in 80 and 160 mg/kg males were significantly increased compared to the vehicle controls. The incidences of hepatocellular neoplasms were significantly greater in dosed groups of female mice than in the vehicle control group. The incidences of hepatocellular neoplasms in all dosed groups of males and females exceeded the historical control ranges. Nonneoplastic hepatocyte changes were seen only in dosed male and female mice. Changes consisted of cytoplasmic alteration and syncytial alteration. The incidences of renal tubule adenoma in males occurred with a positive trend; however, the incidences of carcinoma and hyperplasia did not follow this pattern. An extended evaluation of kidney step sections revealed additional adenomas and hyperplasias in all dosed groups. The combined analysis of single and step sections indicated a dose-related increase in the incidences of renal tubule hyperplasia and renal tubule adenoma or carcinoma (combined), and an increase in the incidences of renal tubule adenoma in male mice. Incidences of thyroidthyroid gland follicular cell hyperplasia were increased in dosed male and female mice compared to vehicle controls. Hyperkeratosis, acanthosis, and exudate were treatment-related changes in the skin at the site of application. The incidences of hyperkeratosis were significantly greater than those in the vehicle control groups in all dosed groups except 40 mg/kg females. GENETIC TOXICOLOGY: Diethanolamine was not mutagenic in any of four strains of Salmonella typhimurium, in the presence or absence of S9 metabolic activation enzymes. No induction of trifluorothymidine resistance was observed in L5178Y mouse lymphoma cells treated with diethanolamine with or without S9. Diethanolamine did not induce significant sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells, with or without S9. Peripheral blood samples collected from male and female mice exposed to 80 to 1,250 mg/kg diethanolamine dermally for 13 weeks showed no increase in micronucleated normochromatic erythrocytes. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of diethanolamine in male F344/N rats administered 16, 32, or 64 mg/kg diethanolamine or in female F344/N rats administered 8, 16, or 32 mg/kg. There was clear evidence of carcinogenic activity of diethanolamine in male and female B6C3F1 mice based on increased incidences of liver neoplasms in males and females and increased incidences of renal tubule neoplasms in males. Dermal administration of diethanolamine to rats was associated with increased incidences of acanthosis (males only), hyperkeratosis, and exudate of the skin and increased incidences and severities of nephropathy in females. Dermal administration of diethanolamine to mice was associated with increased incidences of cytoplasmic alteration (males only) and syncytial alteration of the liver, renal tubule hyperplasia (males only), thyroid gland follicular cell hyperplasia, and hyperkeratosis of the skin. Synonyms: Bis-2-hydroxyethylamine; DEA, diethylolamine; 2,2' dihydroxydiethylamine; diolamine; 2,2'-iminobisethanol; 2,2'-iminodiethanol; iminodiethanol PMID- 12571686 TI - NTP Toxicology and Carcinogenesis Studies of 1-Chloro-2-propanol (Technical Grade) (CAS NO. 127-00-4) in F344/N Rats and B6C3F1 Mice (Drinking Water Studies. AB - 1-Chloro-2-propanol and its positional isomer, 2-chloro-1-propanol, are used as chemical intermediates for the manufacture of propylene oxide, a starting material for production of polyurethane polyols and propylene glycol. The National Cancer Institute nominated 1-chloro-2-propanol for study because of potential for human exposure due to its residues in various foods that are fumigated with ethylene oxide or propylene oxide. Male and female F344/N rats and B6C3F1 mice were exposed to technical grade 1-chloro-2-propanol (75% to 76%% 1 chloro-2-propanol; 24% to 25%% 2-chloro-1-propanol) in drinking water for 14 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, Drosophila melanogaster, and mouse peripheral blood erythrocytes. Continuous breeding studies were conducted in Sprague-Dawley rats. 14-DAY STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were administered 1-chloro-2-propanol in drinking water at concentrations of 0, 100, 330, 1,000, 3,300, or 10,000 ppm for 14 days. Two 10,000 ppm females died before the end of the study. The final mean body weights and body weight gains of 3,300 and 10,000 ppm rats were significantly less than those of the controls; rats in the 10,000 ppm groups lost weight. Water consumption by the 3,300 and 10,000 ppm groups was significantly less than that by the controls throughout the study. The thymus weights of 10,000 ppm rats were significantly less than those of the controls. Exposure to 1-chloro-2-propanol caused cytoplasmic alteration and degeneration of the acinar cells and fatty change in the pancreas, atrophy of the bone marrow, and atrophy and hematopoiesis of the spleen in males and females. 14-DAY STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were administered 1-chloro-2-propanol in drinking water at concentrations of 0, 100, 330, 1,000, 3,300, or 10,000 ppm for 14 days. One male mouse in the 10,000 ppm group died before the end of the study. Mean body weight gains of 10,000 ppm mice were significantly less than those of the controls. Water consumption by 3,300 and 10,000 ppm males and females was significantly less than that by the controls throughout the study. Liver weights of 1,000, 3,300, or 10,000 ppm males and females were significantly greater and thymus weights of 10,000 ppm mice were significantly less than those of the controls. Exposure to 1-chloro-2-propanol caused hepatocellular vacuolization, cytoplasmic alteration and degeneration of the pancreas acinar cells, and atrophy of the spleen in males and females. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were administered 1-chloro-2-propanol at concentrations of 0, 33, 100, 330, 1,000, or 3,300 ppm (equivalent to average daily doses of approximately 5, 10, 35, 100, or 220 mg/kg) for 14 weeks. All rats survived to the end of the study. Mean body weight gains of 3,300 ppm rats were significantly less than those of the controls. Water consumption by the 3,300 ppm male and female rats was significantly less than that by the controls. A minimal to mild anemia was observed in exposed female rats. The cauda epididymis and epididymis weights of 3,300 ppm males were significantly less than those of the controls. The percentage of abnormal sperm in 3,300 ppm males and the concentration of epididymal sperm in 330 ppm males were significantly increased compared to the controls. Kidney and liver weights of males and females exposed to 100 ppm or more were generally greater than those of the controls. The incidences of acinar cell degeneration and fatty change of the pancreas in 1,000 and 3,300 ppm rats, hepatocytic metaplasia of the pancreatic islets in 3,300 ppm females, cytoplasmic vacuolization of the liver in 100, 1,000 and 3,300 ppm males, and renal tubule epithelium regeneration in 3,300 ppm females were increased compared to the controls. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were administered 1-chloro-2-propanol in drinking water at concentrations of 0, 33, 100, 33entrations of 0, 33, 100, 330, 1,000, or 3,300 ppm (equivalent to average daily doses of approximately 5, 15, 50, 170, or 340 mg/kg to males and 7, 20, 70, 260, or 420 mg/kg to females) for 14 weeks. One 330 ppm male died before the end of the study. Mean body weight gains of exposed groups were similar to those of the controls. A minimal anemia was observed in 3,300 ppm males. The right epididymis weight of 3,300 ppm males was significantly greater than that of the controls. Kidney weights of 3,300 ppm mice, liver weights of 1,000 ppm males and of all exposed groups of females, and thymus weights of 1,000 and 3,300 ppm females were greater than those of the controls. The incidences of pancreatic acinar cell degeneration and fatty change in 3,300 ppm males and females and cytoplasmic vacuolization of the liver in all groups of exposed females were significantly increased compared to the controls. The severities of renal tubule cytoplasmic vacuolization were greater in 1,000 and 3,300 ppm males than in the controls. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were administered drinking water containing 0, 150, 325, or 650 ppm 1-chloro-2 propanol (equivalent to average daily doses of approximately 15, 30, or 65 mg/kg during the first several months of the study and 8, 17, or 34 mg/kg for the remainder of the 2-year study) for up to 105 weeks. Survival of all exposed groups was similar to that of the controls. Mean body weights of exposed rats were generally similar to those of the controls throughout most of the study. Water consumption by all exposed groups was similar to that by the controls. No treatment-related neoplasms or nonneoplastic lesions were observed in this study. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were administered drinking water containing 0, 250, 500, or 1,000 ppm 1-chloro-2 propanol (equivalent to average daily doses of approximately 45, 75, or 150 mg/kg to males and 60, 105, or 210 mg/kg to females during the first several months of the study and 25, 50, or 100 mg/kg for the remainder of the 2-year study) for up to 105 weeks. Survival of all exposed groups was similar to that of the controls. The mean body weights of all exposed mice were generally similar to those of the controls throughout the study. Water consumption by all exposed groups was similar to that by the controls. No treatment- related neoplasms or nonneoplastic lesions were observed in this study. GENETIC TOXICOLOGY: 1-Chloro-2-propanol is a demonstrated mutagen in vitro. It was weakly mutagenic in S. typhimurium strain TA100 in the presence of hamster or rat liver S9 activation enzymes and was positive, with and without S9, in TA1535. No mutagenic activity was detected in strains TA97, TA98, and TA1537, with or without S9. In cytogenetic tests with Chinese hamster ovary cells, 1-chloro-2-propanol induced high levels of sister chromatid exchanges and chromosomal aberrations in the presence and the absence of S9. The marked ability of 1-chloro-2-propanol to induce chromosomal effects in vitro was not seen in vivo. Positive results were obtained in a test in D. melanogaster for induction of sex-linked recessive lethal mutations in germ cells of males administered 1-chloro-2-propanol via injection; however, negative results were obtained when males were administered 1-chloro-2-propanol in feed. A subsequent germ cell reciprocal translocation test in D. melanogaster yielded negative results. Further, no induction of micronucleated erythrocytes was observed in peripheral blood of male and female mice administered 1-chloro-2 propanol via drinking water for 14 weeks. CONCLUSIONS: Under the conditions of these 2-year drinking water studies, there was no evidence of carcinogenic activity of technical grade 1-chloro-2-propanol in male or female F344/N rats exposed to 150, 325, or 650 ppm. There was no evidence of carcinogenic activity of technical grade 1-chloro-2-propanol in male or female B6C3F1 mice exposed to 250, 500, or 1,000 ppm. Synonyms: 1-Chloro-2-hydroxypropane, 1-chloroisopropyl alcohol, propylene-α-chlorohydrin, sec-propylene chlorohydrin PMID- 12571687 TI - NTP Toxicology and Carcinogenesis Studies of Primidone (CAS No. 125-33-7) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Primidone is used alone or with other anticonvulsants in the control of grand mal, psychomotor, and focal epileptic seizures. It may control grand mal seizures refractory to other anticonvulsant therapy. Primidone was nominated by the National Cancer Institute for 2-year toxicology and carcinogenicity studies due to its human use as an anticonvulsant. Male and female F344/N rats and B6C3F1 mice received primidone (greater than 99% pure) in feed for 14 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and mouse bone marrow cells. 14-DAY STUDY IN RATS: Five male and five female rats were exposed to 0, 1,250, 2,500, 5,000, 10,000 or 20,000 ppm primidone (equivalent to average daily doses of approximately 120, 240, 500, 970, or 1,100 mg primidone/kg body weight to males and 120, 240, 500, or 900 mg/kg to females) in feed for 14 days. All 20,000 ppm females died before the end of the study as did one 10,000 ppm male and two 20,000 ppm males. The mean body weights of 10,000 ppm males and females and 20,000 ppm males were significantly less than those of the controls. Feed consumption by all exposed rats was generally similar to that by the controls. Males and females in the 10,000 and 20,000 ppm groups were observed to have eye discharge, ataxia, and abnormal posture and were thin and lethargic. 14-DAY STUDY IN MICE: Five male and five female mice were exposed to 0, 625, 1,250, 2,500, 5,000 or 10,000 ppm primidone (equivalent to average daily doses of approximately 100, 200, 400, or 800 mg/kg body weight to males and 100, 250, 500, or 900 mg/kg to females) in feed for 14 days. All mice in the 10,000 ppm groups and one male and one female mouse in the 5,000 ppm groups died on day 3 of the study. The mean body weights of mice in the 625, 1,250, 2,500, and 5,000 ppm groups were similar to those of the controls. Feed consumption by all exposed mice was generally similar to that by the controls. Males and females in the 10,000 ppm groups were observed to have abnormal posture, ataxia, and lethargy. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to 0, 300, 600, 1,300, 2,500, or 5,000 ppm primidone (equivalent to average daily doses of approximately 20, 40, 100, 200, or 400 mg/kg) in feed for 14 weeks. All rats survived to the end of the study. The mean body weights of male and female rats in the 2,500 and 5,000 ppm groups were significantly less than those of the controls. Feed consumption by all exposed rats was generally similar to that by the controls. A minimal to mild exposure-related thrombocytosis occurred on day 22 and at week 14 in all exposed groups of male rats and in females in the 1,300 ppm or greater groups. A minimal decrease in hemoglobin concentration occurred in 2,500 and 5,000 ppm male and female rats on day 22 and at week 14. The incidences of centrilobular hepatocyte hypertrophy in male rats exposed to 600 ppm or greater and in female rats exposed to 1,300 ppm or greater were significantly greater than those in the controls. The severity of chronic nephropathy in male rats exposed to 1,300 ppm or greater increased with increasing exposure concentration. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were exposed to 0, 300, 600, 1,300, 2,500, or 5,000 ppm primidone (equivalent to average daily doses of approximately 50, 100, 200, 400, or 1,000 mg/kg to males and 60, 120, 220, 440, or 1,100 mg/kg to females) in feed for 14 weeks. Three male and two female mice in the 5,000 ppm group died during week 1 of the study. The final mean body weights of all exposed groups were similar to those of the controls. Feed consumption by male mice in the 5,000 ppm group was slightly greater than that by the controls; this may have been due to feed spillage. Male and female mice in the 5,000 ppm groups were ataxic and lethargic. Compared to controls, the estrous cycle lengths of females exposed to 1,300, 2,500, or 5,000 ppm were significantly longer. The liver weights of male and female mice exposed to 600 po 600 ppm or greater were significantly greater than those of the controls. The incidences of centrilobular hepatocyte hypertrophy in all exposed males and in females exposed to 600 ppm or greater and the incidences of cytoplasmic alteration of the adrenal gland and hematopoietic cell proliferation of the spleen in 2,500 and 5,000 ppm males and in 5,000 ppm females were significantly greater than in the controls. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to 0, 600, 1,300, or 2,500 ppm primidone (equivalent to average daily doses of approximately 25, 50, or 100 mg/kg) in feed for 2 years. Survival, Body Weights, and Feed Consumption Survival of the 1,300 and 2,500 ppm males was sig nificantly less than that of the controls. The mean body weights of males and females in the 2,500 ppm groups were less than those of the controls, beginning at week 29 for males and week 17 for females; the mean body weights of 1,300 ppm males and females were less than those of the controls during the second year of the study. Feed consumption by all exposed groups of rats was generally similar to that by the controls. Pathology Findings Male rats exposed to primidone had increased inci dences of thyroid gland follicular cell neoplasms (adenoma and/or carcinoma). All exposed groups of male rats had follicular cell adenomas or carcinomas (combined) at incidences above the historical control range, with the highest incidence in the 1,300 ppm group. Hepatocyte cytoplasmic vacuolation and centrilobular hypertrophy were associated with primidone exposure in male and female rats. These changes were more severe in females than in males and the incidences in all exposed groups of females were significantly greater than those in the controls. Females in the 2,500 ppm group had an increased incidence of hepatocellular eosinophilic foci. In 2,500 ppm males, the incidence of renal tubule hyperplasia was greater than that in the controls in the standard evaluation. Additional hyperplasias were found in the extended evaluation, and the incidences in exposed groups of males were significantly greater than that in the controls. In the extended evaluation, the incidence of renal tubule adenoma in 2,500 ppm males was significantly increased. The incidence of adenoma or carcinoma (combined) in 2,500 ppm males in the combined standard and extended evaluations were marginally increased over those in the controls. Male rats had an exposure-related increase in the severity of chronic nephropathy, which probably accounted for the reduced survival in the 1,300 and 2,500 ppm groups. The incidences of kidney cysts were increased in 1,300 and 2,500 ppm males. Hyperparathyroidism, secondary to the loss of renal function, was present in many exposed male rats. The incidences of parathyroid gland hyperplasia in all groups of exposed males were significantly greater than that in the controls. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed to dietary levels of 0, 300, 600, or 1,300 ppm primidone (equivalent to average daily doses of approximately 30, 65, or 150 mg/kg to males and 25, 50, or 100 mg/kg to females) in feed for 2 years. Survival, Body Weights, Feed Consumption, and Clinical Findings Survival of the 1,300 ppm males was significantly less than that of the controls. During the second year of the study, the mean body weights of 1,300 ppm male and female mice were less than those of the controls. The final mean body weights of 600 ppm males and females were less than those of the controls. Feed consumption by all exposed groups of mice was similar to that by the controls. During the latter part of the study, a treatment-related increase in the number of animals with swelling of the abdominal area was observed; necropsy revealed that the swelling was due to liver nodules/masses. Pathology Findings The liver was a target organ in both male and female mice. The incidences and multiplicities of hepatocellular neoplasms (hepatocellular adenoma, hepatocellular carcinoma, and hepatoblastoma) in all exposed groups of males and females (except hepatoblastoma in females) were significantly greater than those in the controls. The incidences of hepatocellular adenoma or carcinoma (combined) and hepatocellular adenoma, hepatocellular carcinoma, or hepatoblastoma (combined) in all exposed groups exceeded the historical control ranges in 2-year NTP studies. The incidences of centrilobular hepatocyte hypertrophy were increased in exposed groups of males and females, and the severities increased with increasing exposure concentration. The incidences of cytoplasmic vacuolization were increased in all exposed groups of females and in 300 ppm males. Incidences of eosinophilic focus in all exposed groups of females were significantly greater than those in the controls. Proliferative changes occurred in the thyroid gland in an exposure-related manner in male and female mice. Incidences of follicular cell hyperplasia were increased in all exposed groups of males and in 600 and 1,300 ppm females, but incidences of follicular cell adenomas were increased only in male mice. GENETIC TOXICOLOGY: Primidone was mutagenic in Salmonella typhimurium strain TA1535 in the absence of S9 activation only; no mutagenicity was detected in strain TA98, TA100, or TA1537, with or without S9. Primidone did not induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells, with or without S9. The single in vivo study with primidone, a mouse bone marrow micronucleus test, also gave negative results. CONCLUSIONS: Under the conditions of these 2-year feed studies, there was equivocal evidence of carcinogenic activity of primidone in male F344/N rats based on a marginal increase in thyroid gland follicular cell neoplasms, primarily adenomas, and a marginal increase in renal tubule neoplasms. There was no evidence of carcinogenic activity of primidone in female F344/N rats exposed to 600, 1,300, or 2,500 ppm. There was clear evidence of carcinogenic activity of primidone in male B6C3F1 mice based on the increased incidences of hepatocellular neoplasms, and the increased incidence of thyroid gland follicular cell adenomas was also considered to be chemical related. There was clear evidence of carcinogenic activity of primidone in female B6C3F1 mice based on the increased incidences of hepatocellular neoplasms. Exposure of rats to primidone resulted in increased incidences of hepatocyte cytoplasmic vacuolization and centrilobular hypertrophy in males and females and eosinophilic foci in females. The increased severity of nephropathy and increased incidence of renal tubule hyperplasia in male rats were related to primidone exposure. Exposure of male mice to primidone resulted in hepatocyte centrilobular hypertrophy and thyroid gland follicular cell hyperplasia. Exposure of female mice to primidone resulted in hepatocyte centrilobular hypertrophy and cytoplasmic vacuolization, eosinophilic focus, and thyroid gland follicular cell hyperplasia. Synonyms: 5 Aethyl-5-phenyl-hexahydropyrimidin-4,6-dion; 2-deoxyphenobarbital; 2 desoxyphenobarbital; desoxyphenobarbitone; 5-ethyldihydro-5-phenyl-4,6 (1H,5H) pyrimidinedione; 5-ethylhexahydro-4,6-dioxo-5-phenylphrimidine; 5-ethylhexahydro 5-phenylpyrimidine-4,6-dione; 5-ethyl-5-phenylhexahydropyrimidine-4,6-dione Trade names: Cyral; Hexadiona; Hexamidine; Lepimidin; Lepsiral; Majsolin; Midone; Milepsin; Misodine; Misolyne; Mizodin; Mizolin; Mylepsin; Mylepsinum; Mysedon; Mysoline; Prilepsin; Primacione; Primaclone; Primacone; Primakton; Primadon; Prysoline; Pyrimidone; ROE 101; Sertan PMID- 12571688 TI - NTP Toxicology and Carcinogenesis Studies of Tetrahydrofuran (CAS No. 109-99-9) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Tetrahydrofuran is used as a reaction medium for Grignard and metal hydride reactions; in the synthesis of butyrolactone, succinic acid, and 1,4-butanediol diacelate; in the fabrication of articles for packaging, transporting, and storing of foods; as a solvent for dyes and lacquers; and as a chemical intermediate in polymerization solvent for fat oils, unvulcanized rubber, resins, and plastics. Tetrahydrofuran is also an indirect food additive when it is in contact with the surface of articles intended for use in food processing. Tetrahydrofuran was nominated for study because of the potential for occupational exposure in humans. Male and female F344/N rats and B6C3F1 mice were exposed to tetrahydrofuran (approximately 99% pure) by inhalation for 13 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, Drosophila melanogaster, mouse bone marrow cells, and mouse peripheral blood cells erythrocites. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to 0 (chamber control), 66, 200, 600, 1,800, or 5,000 ppm tetrahydrofuran by inhalation, 6 hours per day, 5 days per week, for 14 weeks. All rats survived until the end of the study. Final mean body weights and mean body weight gains of exposed groups of male and female rats were similar to those of the chamber controls. Immediately after exposure, male and female rats in the 5,000 ppm groups exhibited ataxia. Hematologic and serum chemistry changes were minimal, with most values falling within physiologic ranges. Absolute and relative thymus and spleen weights of male and female rats exposed to 5,000 ppm were significantly less than those of the chamber controls. Absolute and relative liver weights of female rats exposed to 5,000 ppm were significantly greater than those of the chamber controls. Increased incidences of minimal to mild hyperplasia of the forestomach were observed in male and female rats exposed to 5,000 ppm. Minimal suppurative inflammation was associated with forestomach hyperplasia in two male rats and four female rats exposed to 5,000 ppm. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were exposed to 0, 66, 200, 600, 1,800, or 5,000 ppm tetrahydrofuran by inhalation, 6 hours per day, 5 days per week, for 14 weeks. Two male mice exposed to 5,000 ppm died during weeks 2 and 8 of the study; one male mouse from the 5,000 ppm group was killed in a moribund state during week 4. All female mice survived until the end of the study. The final mean body weights and mean body weight gains of all exposed groups of male mice were similar to those of the chamber controls. The final mean body weight and mean body weight gain of the 5,000 ppm female mice were significantly greater than those of the chamber controls. Male and female mice exposed to 1,800 or 5,000 ppm were observed in a state of narcosis (described by stupor) during exposure periods. Mice exposed to 1,800 ppm were fully awake and alert immediately after exposure; however, mice exposed to 5,000 ppm required up to 2 hours for recovery. Absolute and relative liver weights of male mice exposed to 600 ppm or greater and of female mice exposed to 1800 or 5,000 ppm were significantly greater than those of the chamber controls. Absolute and relative thymus weights of male mice exposed to 600, 1,800, or 5,000 ppm were significantly less than those of the chamber controls. The incidences of minimal to mild centrilobular cytomegaly of the liver in male and female mice exposed to 5,000 ppm were significantly greater than those in the chamber controls. The adrenal glands of all female mice exposed to 5,000 ppm had mild degeneration of the X-zone of the innermost cortex. Uterine atrophy was observed in all female mice exposed to 5,000 ppm. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed to 0, 200, 600, or 1,800 ppm tetrahydrofuran by inhalation, 6 hours per day, 5 days per week, for 105 weeks. Survival, Body Weights, and Clinical Findings Survival and mean body weights of male and femand female rats exposed to tetrahydrofuran were similar to those of the chamber controls. Pathology Findings: The incidences of renal tubule epithelial adenoma or carcinoma (combined) in exposed males occurred with a positive trend, and the incidences in 600 and 1,800 ppm males exceeded the historical range for chamber controls in 2-year NTP inhalation studies. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed to 0, 200, 600, or 1,800 ppm tetrahydrofuran by inhalation, 6 hours per day, 5 days per week, for 105 weeks. Survival, Body Weights, and Clinical Findings After week 36, the survival of male mice exposed to 1,800 ppm was significantly less than that of the chamber controls. Mean body weights of male and female mice exposed to tetrahydrofuran were similar to those of the chamber controls throughout the study. Male mice exposed to 1,800 ppm were observed to be in a state of narcosis during and up to 1 hour after the exposure periods. Pathology Findings: The incidences andmultiplicity of hepatocellular neoplasms were significantly greater in female mice exposed to 1,800 ppm than in the chamber controls. The incidence of nephropathy in 200 ppm male mice was significantly greater than that in the chamber control group. Male mice exposed to 1,800 ppm had significantly greater incidences of nonneoplastic lesions of the urogenital tract than did the chamber controls. The incidences of inflammation of the penis and urethra and necrosis of the urethra in 1,800 ppm males were slightly greater than those in the chamber controls; these may have been secondary effects of ascending urinary tract infection. GENETIC TOXICOLOGY: Tetrahydrofuran showed little evidence of mutagenic activity in a variety of in vitro and in vivo assays. It was not mutagenic in Salmonella typhimurium, and it did not induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells. These in vitro tests were conducted with and without exogenous metabolic activation from induced liver S9 enzymes. No increase in sex linked recessive lethal mutations was detected in germ cells of male D. melanogaster exposed to tetrahydrofuran by feeding or injection. Results of in vivo assays for induction of chromosomal aberrations and sister chromatid exchanges in mouse bone marrow cells were negative. A micronucleus test in male and female mice exposed to tetrahydrofuran for 14 weeks showed no significant increases in the frequency of micronucleated erythrocytes in peripheral blood of female mice, but in male mice, analysis of micronucleated normochromatic erythrocyte levels revealed a small increase above baseline that was concluded to be equivocal. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was some evidence of carcinogenic activity of tetrahydrofuran in male F344/N rats based on increased incidences of renal tubule adenoma or carcinoma (combined). There was no evidence of carcinogenic activity of tetrahydrofuran in female F344/N rats exposed to 200, 600, or 1,800 ppm or male B6C3F1 mice exposed to 200, 600, or 1,800 ppm. There was clear evidence of carcinogenic activity of tetrahydrofuran in female B6C3F1 mice based on increased incidences of hepatocellular neoplasms. Synonyms: Butylene oxide; cyclotetramethylene oxide; diethylene oxide; 1,4-epoxybutane; furanidine; hydrofuran; oxacyclopentane; oxolane; tetramethylene oxide PMID- 12571689 TI - NTP Toxicology and Carcinogenesis Studies of Isoprene (CAS No. 78-79-5) in F344/N Rats (Inhalation Studies). AB - Isoprene, the monomeric unit of natural rubber and naturally occurring terpenes and steroids, is primarily obtained as a by-product of naphtha cracking for ethylene production. It is emitted from plants and trees, has been detected in tobacco smoke and automobile exhaust, and was identified as a major endogenous hydrocarbon in human breath. Isoprene was selected for toxicologic evaluation because of its structural similarity to 1,3-butadiene, a potent, multi-organ, rodent carcinogen, and the potential for human exposure due to its large annual production volume. A previous 26-week inhalation study followed by a 26-week recovery period provided clear evidence of carcinogenic activity of isoprene in male B6C3F1 mice. A similar study in male F344/N rats was inconclusive. Male and female F344/N rats were exposed to isoprene (99% pure) by whole body inhalation for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, mouse bone marrow and peripheral blood cells, and rat lung fibroblasts. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to 220, 700, or 7,000 ppm isoprene by inhalation, 6 hours per day, 5 days per week, for 105 weeks. Survival and Body Weights: Survival rates and mean body weights of exposed male and female rats were similar to those of the chamber controls throughout the study. Urinary Vinyl Lactic Acid Biomarker of Exposure: At 3, 6, 12, and 18 months, the concentrations of vinyl lactic acid normalized to creatinine in the urine increased with increasing exposure concentration in all exposed groups of male and female rats; however, these increases were not proportional to isoprene exposure concentrations, indicating nonlinear metabolism over this range of exposure concentrations. Pathology Findings: Exposure-related increases in the incidences of mam mary gland fibroadenoma were observed in male rats in all groups. The incidences of fibroadenoma in 7,000 ppm males and in all groups of exposed females were significantly greater than those in the chamber control groups. The incidences of fibroadenoma in all exposed groups of males and females and of multiple fibroadenoma in 7,000 ppm males and in all groups of exposed females exceeded the historical control ranges. In addition, the finding of mammary gland carcinoma in exposed male rats was noteworthy because this neoplasm rarely occurs in control male rats. The incidences of renal tubule adenoma in 700 and 7,000 ppm males and of renal tubule hyperplasia in 7,000 ppm males were significantly greater than those in the chamber controls. The severity of kidney nephropathy was slightly increased in 7,000 ppm males when compared to chamber controls. An exposure related increase in the incidences of interstitial cell adenoma of the testis was observed in male rats. The incidences of bilateral interstitial cell adenoma and of unilateral and bilateral interstitial cell adenoma (combined) of the testis in 700 and 7,000 ppm males were significantly greater than those in the chamber controls. The incidences of interstitial cell adenoma in 700 and 7,000 ppm males exceeded the historical control range. Several rare neoplasms including benign astrocytoma, malignant glioma, malignant medulloblastoma be nign meningeal granular cell tumor, and meningeal sarcoma were observed in the brain of exposed female rats. These neoplasms have seldom or never occurred in historical chamber controls. The incidences of splenic fibrosis in 700 and 7,000 ppm males were significantly greater than that in the chamber control group. GENETIC TOXICOLOGY: Isoprene was not mutagenic in S. typhimurium and did not induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells with or without exogenous metabolic activation; however, in mice, isoprene induced increases in the frequency of sister chromatid exchanges in bone marrow cells and in the frequency of micronucleated erythrocytes in peripheral blood. The cell cycle duration of proliferating bone marrow cells of mice f mice exposed to 7,000 ppm isoprene was significantly lengthened. No increases in the frequency of chromosomal aberrations were observed in bone marrow cells of male mice after 12 days of exposure to isoprene, and lung fibroblasts of male and female rats exposed to isoprene for 4 weeks showed no increase in the frequency of micronuclei. CONCLUSIONS: Under the conditions of this 2-year inhalation study, there was clear evidence of carcinogenic activity of isoprene in male F344/N rats based on increased incidences of mammary gland fibroadenoma and carcinoma, renal tubule adenoma, and testicular interstitial cell adenoma. There was some evidence of carcinogenic activity of isoprene in female F344/N rats based on increased incidences and multiplicity of mammary gland fibroadenoma. A low incidence of rare brain neoplasms in exposed female rats may have been due to exposure to isoprene. Exposure to isoprene by inhalation for 2 years resulted in increased incidences of renal tubule hyperplasia and splenic fibrosis in male rats. Synonyms: Isopentadiene; b-methylbivinyl; 2-methyl-1,3-butadiene PMID- 12571690 TI - Who said that? PMID- 12571691 TI - Drug interactions: gasoline and matches. PMID- 12571692 TI - Guidelines for implantable cardioverter defibrillator follow-up in Canada: a consensus statement of the Canadian Working Group on Cardiac Pacing. AB - A survey on the follow-up practices of patients with implantable cardioverter defibrillators (ICDs) in Canada was conducted by the Canadian Working Group on Cardiac Pacing in 1999. The survey identified the need for national guidelines. The present guidelines for ICD follow-up represent a consensus statement of the Canadian Working Group on Cardiac Pacing. They recommend that patients be assessed before hospital discharge, at two to 12 weeks following implantation, six months after implantation, and at a minimum of every six months thereafter. More frequent assessments may be required for some patients depending on associated cardiovascular problems and specific ICD devices. A typical follow-up visit should include a targeted cardiovascular assessment, interrogation of the ICD, review of telemetered data, assessment of the underlying rhythm, pacing and sensing thresholds, lead impedance and appropriate reprogramming of ICD parameters to optimize device function and longevity. PMID- 12571693 TI - CCORT/CCS quality indicators for acute myocardial infarction care. AB - BACKGROUND: Although quality indicators for the care of acute myocardial infarction (AMI) patients have been described for other countries, there are none specifically designed for the Canadian health care system. The authors' goal was to develop a set of Canadian quality indicators for AMI care. METHODS: A literature review identified existing quality indicators for AMI care. A list of potential indicators was assessed by a nine-member panel of clinicians from a variety of disciplines using a modified-Delphi panel process. After an initial round of rating the potential indicators, a series of indicators was identified for a second round of discussion at a national meeting. Further refinement of indicators occurred following a teleconference and review by external reviewers. RESULTS: To identify an AMI cohort, case definition criteria were developed, using a hospital discharge diagnosis for AMI of International Classification of Diseases-Ninth revision (ICD-9) code 410.x. Thirty-seven indicators for AMI care were established. Pharmacological process of care indicators included administration of acetylsalicylic acid, beta-blockers, angiotensin-converting enzyme inhibitors, thrombolytics and statins. Mortality and readmissions for AMI, unstable angina and congestive heart failure were recommended as outcome indicators. Nonpharmacological indicators included median length of stay in the emergency department, and median waiting times for cardiac catheterization, percutaneous coronary intervention and/or coronary artery bypass graft surgery. INTERPRETATION: A set of Canadian quality indicators for the care of AMI patients has been established. It is anticipated that these indicators will be useful to clinicians and researchers who want to measure and improve the quality of AMI patient care in Canada. PMID- 12571694 TI - Is outpatient coronary angioplasty and stenting feasible and safe? Results of a retrospective analysis. AB - OBJECTIVE: To assess the safety of coronary angioplasty with or without stenting and with same day discharge in a population of outpatients or referred patients. This study also assessed the changing practice in the access route and the increasing use of stents. METHODS AND RESULTS: In this retrospective and descriptive study, the authors reviewed the files of ambulatory or referred patients hospitalized within seven days of a heart catheterization with coronary angiography between January 1997 and December 1999. Overall, 1856 patients were included, of which 546 (29.4%) had a coronary angioplasty. In the first six months, stents were used in 13.3% of outpatients and 23.5% of referred patients, while in the last six months 75.0% of outpatients and 73.0% of referred patients were stented. A total of 156 patients were triaged for a prolonged observation period of which 66 (12.1%) received some treatment (heparin, abciximab, coronary artery bypass graft or transfusion). Two patients (0.4%) had acute thrombosis. No patients experienced complications that could have been prevented by a one or two day routine observation period. While 47% of patients were accessed by the radial artery in the first six months, 77.2% of patients were accessed by the radial artery in the last six months. Significant access complications occurred in 11 patients (0.6%). CONCLUSION: Outpatient angioplasty is feasible and, according to the retrospective analysis of the authors' admittedly incomplete data, safe. The use of stents facilitates same-day discharge. Pre-, per- and postprocedural triage allows identification of high risk patients for a longer observation period with or without additional treatment. PMID- 12571695 TI - Prevalence of metabolic alterations predictive of cardiovascular disease risk in the Quebec population. AB - BACKGROUND: It is well established that cardiovascular diseases (CVD) are the most important cause of morbidity and mortality in North America and in industrialized countries. Furthermore, the presence of CVD risk factors is widespread in Canada, and two-thirds of adult Canadians present at least one traditional risk factor for CVD. OBJECTIVE: To quantify the prevalence of metabolic alterations predictive of an increased risk of CVD in a representative sample of the Quebec population. METHODS: A representative sample of the Quebec population was obtained through the Quebec Health Survey 1990. The cohort of 1844 subjects (907 men and 937 women), aged from 18 to 74 years, completed the evaluation, which consisted of interviews, validated questionnaires for personal and family history of CVD risk factors, and a physical examination. Anthropometric measurements, blood pressure, fasting plasma lipoprotein-lipid levels as well as fasting glucose and insulin concentrations were obtained. RESULTS: Thirty one per cent of the Quebec population had a normal body weight defined by body mass index (BMI) less than 25 kg/m2 with a healthy metabolic profile (glycemia less than 6.1 mmol/L, total cholesterol less than 5.2 mmol/L, low density lipoprotein-cholesterol less than 3.4 mmol/L, triglycerides less than 2.3 mmol/L and high density lipoprotein-cholesterol greater than 0.90 mmol/L). On the other hand, 28% of the population was treated for known metabolic chronic diseases (CVD, diabetes, dyslipidemias and hypertension), whereas the remaining 33% of the Quebec population (BMI greater than 25 kg/m2) were characterized by an altered metabolic profile predictive of an increased CVD risk. Furthermore, 75% of men and 70% of women with an altered metabolic profile presented at least two CVD risk factors. Overall, women tended to display a significantly healthier metabolic risk profile than men (P<0.05). CONCLUSIONS: According to our results, only 31% of the Quebec population is nonobese without major alterations in their metabolic risk profile. Indeed, 61% of Quebecers are either characterized by chronic diseases or by an altered metabolic profile increasing their risk for the development of CVD and type 2 diabetes. Such a high prevalence should be a source of concern justifying the development of simple screening tools for early identification of the asymptomatic carriers of this cluster of metabolic abnormalities. PMID- 12571696 TI - The fate of endothelial function testing: rationale and design of the Firefighters And Their Endothelium (FATE) study. AB - Endothelial dysfunction plays a pivotal role in the development and progression of atherosclerotic vascular disease. The endothelium is strategically located between blood and vascular smooth muscle, making it both vulnerable to a variety of injurious stimuli but also available for interrogation as a marker of vascular health. Firefighters And Their Endothelium (FATE) is a prospective, longitudinal cohort study designed to assess the relationship between endothelial function, emerging cardiovascular risk factors and ultimately atherosclerotic vascular disease. It is hypothesized that participants with impaired endothelial function will be at increased risk of atherosclerotic complications. This Canadian initiative will recruit 1600 middle-aged participants with no known history of cardiovascular disease to be followed for cardiovascular events over the next decade. Quantitative B-mode ultrasound will be employed to assess endothelial function and subclinical atherosclerosis. This research is designed to redefine the approach to the primary prevention of atherosclerosis. PMID- 12571697 TI - The angiographic correlation between ST segment depression in noninfarcted leads and the extent of coronary artery disease in patients with acute inferior myocardial infarction: a clue for multivessel disease. AB - BACKGROUND: Although reciprocal ST segment depression (RSTD) in patients with acute inferior myocardial infarction is a common electrocardiogram finding, its significance is not yet established. In this prospective study, the relationship between RSTD and the extent of coronary artery disease (CAD) was investigated. PATIENTS AND METHODS: One hundred eighty-eight patients with acute inferior myocardial infarction who received thrombolytic therapy were enrolled in this study. The magnitude and location of ST segment depression in noninfarcted leads and the maximum ST segment elevation (STEmax) in inferior leads were measured. All patients were divided into two main groups according to the presence of RSTD and five subgroups according to the location of RSTD, the maximum RSTD and the STEmax. The coronary angiography was performed in all patients 28 +/- 4 days after acute myocardial infarction. RESULTS: There were no significant differences in the proportion of coronary disease risk factors in patients with, versus those without, RSTD (P=0.6). Multivessel CAD was present in 63 of the 108 (58%) patients with RSTD and in 32 of the 80 (40%) patients with no RSTD (P=0.02). According to the location of reciprocal changes, multivessel disease was present in significantly more patients with anterior RSTD concomitant with or without lateral ST segment depression (P=0.01 and P=0.03, respectively); the proportion of single vessel disease was greater in patients with only lateral RSTD (P=0.02). In addition, the presence of anterior RSTD to a greater magnitude than the STEmax in inferior myocardial infarction increases the likelihood of multivessel disease (P=0.006). CONCLUSIONS: The presence of RSTD during an acute inferior myocardial infarction correlates with the presence of multivessel CAD and may not be only an electrical phenomenon. PMID- 12571698 TI - Congenital and acquired long QT syndromes. AB - Exploration into the underlying genetic causes of congenital long QT syndrome (LQTS) has opened the door to our understanding of repolarization disorders. Expression of LQTS mutations has led to an improved understanding of the mechanisms of arrhythmogenesis, clinical diagnostic tools and channel specific therapy. Further insight into the mechanisms underlying the more common acquired LQTS is emerging from gene and channel studies that have used the congenital syndrome as a springboard for directing research to improve understanding. This review summarizes the clinical, genetic and electrophysiological understanding of congenital and acquired LQTS. PMID- 12571700 TI - Frontiers in cardiovascular research: forging further scientific collaborations between Canada and the United States. PMID- 12571716 TI - International law and public health. PMID- 12571717 TI - The framework convention on tobacco control. PMID- 12571718 TI - Public health in international law: the contribution of the World Summit on Sustainable Development. PMID- 12571719 TI - Effect of supplementary feeding on the prevention of mild-to-moderate wasting in conditions of endemic malnutrition in Guatemala. AB - OBJECTIVE: To estimate the effect of supplementary feeding on the prevention of wasting in preschool children in a rural area of Guatemala with a high prevalence of malnutrition. METHODS: Children aged 6-48 months, with a weight-for-length exceeding 90% of that of the median NCHS/WHO/CDC reference population, received either Atole, a drink with a high protein and moderate energy content, or Fresco, a drink with no protein and a low energy content. Children consuming amounts of Atole equivalent to more than 10% of the age-adjusted recommended dietary intake for energy were termed "supplemented." A comparable group of children consuming Fresco was termed "non-supplemented". The energy intake in the supplemented group was higher than that in the non-supplemented group by 16-18% of the recommended daily intake. The corresponding difference in protein intake was 45-80% of the recommended daily intake. The children were followed up at intervals of three or six months in order to evaluate their weight-for-length development. The difference between supplemented and non-supplemented children in the proportions maintaining their weight-for-length category at the end of the study intervals represented the prevention of wasting attributable to supplementary feeding (attributable prevention). Households were used as units of analysis in order to avoid cluster effects. FINDINGS: The attributable prevention in children aged 6 24 months with weight-for-length between 90% and 99.9% of the reference population at entry ranged from 0.21 to 0.26 and was statistically significant after three and six months of supplementation. However, it was not significant in children at or above 100% weight-for-length or in older children. Children above 100% weight-for-length did not become overweight as a result of supplementation. CONCLUSION: Supplementary feeding of children aged 6-24 months in populations with inadequate dietary intakes can prevent the onset of wasting in a large proportion of children. PMID- 12571720 TI - Physical activity during leisure and commuting in Tianjin, China. AB - OBJECTIVE: To investigate physical activity during leisure time and commuting among persons aged 15-69 years in the urban population of Tianjin, China, and to assess its associations with demographic and health-related characteristics. METHODS: In 1996 a cross-sectional survey of 2002 males and 1974 females provided information on physical activity during leisure time and commuting and on demographics and health behaviours. FINDINGS: No leisure-time physical activity was engaged in by 67% of females and 61% of males. However, only 4% of females and 9% of males reported an absence of physical activity during commuting. The mean duration of leisure-time physical activity for the whole population was about 10 min per day. The average commuting time on foot or by bicycle was about 30 min. Leisure-time physical activity was more frequent among highly educated people, people with high incomes, white-collar workers, married people, non smokers, or people commuting on foot or by bicycle than among other people. Persons with low incomes, male blue-collar workers and married people were more likely than others to engage in 30 min or more per day of physical activity on foot or by bicycle when commuting. CONCLUSION: People in Tianjin engaged in a high level of physical activity when commuting and a low level of leisure-time physical activity. PMID- 12571721 TI - How human immunodeficiency virus voluntary testing can contribute to tuberculosis control. AB - Human immunodeficiency virus (HIV) is fueling the tuberculosis (TB) epidemic, particularly in sub-Saharan Africa. However, despite their close epidemiological links, the public health responses have largely been separate. WHO has set out a strategy to decrease the burden of HIV-related TB, comprising interventions against both TB and HIV. Voluntary counselling and testing (VCT) for HIV can link TB and HIV programme activities. The benefits of VCT for HIV to TB patients include referral for appropriate clinical care and support for those testing HIV positive. Likewise, people attending a centre for VCT can benefit from TB screening: those found to be both HIV-positive and with active TB need referral for TB treatment; those without active TB should be offered TB preventive treatment with isoniazid. To explore how VCT for HIV can contribute to a more coherent response to TB, WHO is coordinating the ProTEST Initiative. The name "ProTEST" is derived from the Promotion of voluntary testing as an entry point for access to the core interventions of intensified TB case-finding and isoniazid preventive treatment. Other interventions may be added to provide finally a comprehensive range of HIV and TB prevention and care interventions. Under the ProTEST Initiative, pilot districts are establishing links between centres for VCT for HIV and TB prevention and care. This will pave the way for large-scale operationalization of the comprehensive range of interventions needed to control TB in settings with high HIV prevalence. PMID- 12571722 TI - International law and communicable diseases. AB - Historically, international law has played a key role in global communicable disease surveillance. Throughout the nineteenth century, international law played a dominant role in harmonizing the inconsistent national quarantine regulations of European nation-states; facilitating the exchange of epidemiological information on infectious diseases; establishing international health organizations; and standardization of surveillance. Today, communicable diseases have continued to re-shape the boundaries of global health governance through legally binding and "soft-law" regimes negotiated and adopted within the mandate of multilateral institutions - the World Health Organization, the World Trade Organization, the Food and Agriculture Organization, and the Office International des Epizooties. The globalization of public health has employed international law as an indispensable tool in global health governance aimed at diminishing human vulnerability to the mortality and morbidity burdens of communicable diseases. PMID- 12571723 TI - A global response to a global problem: the epidemic of overnutrition. AB - It is estimated that by 2020 two-thirds of the global burden of disease will be attributable to chronic noncommunicable diseases, most of them strongly associated with diet. The nutrition transition towards refined foods, foods of animal origin, and increased fats plays a major role in the current global epidemics of obesity, diabetes and cardiovascular diseases, among other noncommunicable conditions. Sedentary lifestyles and the use of tobacco are also significant risk factors. The epidemics cannot be ended simply by encouraging people to reduce their risk factors and adopt healthier lifestyles, although such encouragement is undoubtedly beneficial if the targeted people can respond. Unfortunately, increasingly obesogenic environments, reinforced by many of the cultural changes associated with globalization, make even the adoption of healthy lifestyles, especially by children and adolescents, more and more difficult. The present paper examines some possible mechanisms for, and WHO's role in, the development of a coordinated global strategy on diet, physical activity and health. The situation presents many countries with unmanageable costs. At the same time there are often continuing problems of undernutrition. A concerted multisectoral approach, involving the use of policy, education and trade mechanisms, is necessary to address these matters. PMID- 12571724 TI - Biomedicine and international human rights law: in search of a global consensus. AB - Global challenges raised by biomedical advances require global responses. Some international organizations have made significant efforts over the last few years to establish common standards that can be regarded as the beginning of an international biomedical law. One of the main features of this new legal discipline is the integration of its principles into a human rights framework. This strategy seems the most appropriate, given the role of "universal ethics" that human rights play in our world of philosophical pluralism. In addition to the general standards that are gradually being established, a widespread consensus exists on the urgency of preventing two specific procedures: human germ line interventions and human reproductive cloning. PMID- 12571725 TI - International law, human rights and HIV/AIDS. AB - This article explores the relevance of international human rights law in the response to the HIV/AIDS epidemic at national and international levels. Public health advocates can use arguments based on this body of law to promote responses to HIV/AIDS that reflect sound public health principles and documented best practice. Development assistance is increasingly linked to rights-based approaches, such as participatory processes, and strategic alliances between health professionals, organizations of people living with HIV/AIDS, and affected communities. Legal and human rights advocacy strategies are increasingly productive and necessary. PMID- 12571726 TI - International environmental law and global public health. AB - The environment continues to be a source of ill-health for many people, particularly in developing countries. International environmental law offers a viable strategy for enhancing public health through the promotion of increased awareness of the linkages between health and environment, mobilization of technical and financial resources, strengthening of research and monitoring, enforcement of health-related standards, and promotion of global cooperation. An enhanced capacity to utilize international environmental law could lead to significant worldwide gains in public health. PMID- 12571727 TI - Global governance, international health law and WHO: looking towards the future. AB - The evolving domain of international health law encompasses increasingly diverse and complex concerns. Commentators agree that health development in the twenty first century is likely to expand the use of conventional international law to create a framework for coordination and cooperation among states in an increasingly interdependent world. This article examines the forces and factors behind the emerging expansion of conventional international health law as an important tool for present and future multilateral cooperation. It considers challenges to effective international health cooperation posed for intergovernmental organizations and other actors involved in lawmaking. Although full consolidation of all aspects of future international health lawmaking under the auspices of a single international organization is unworkable and undesirable, the World Health Organization (WHO) should endeavour to serve as a coordinator, catalyst and, where appropriate, platform for future health law codification. Such leadership by WHO could enhance coordination, coherence and implementation of international health law policy. PMID- 12571728 TI - The Preamble of the Constitution of the World Health Organization. PMID- 12571729 TI - Constitution of the World Health Organization. 1946. PMID- 12571730 TI - How objective are the supporters of the Haemoglobin Colour Scale? PMID- 12571731 TI - Further clarity on vaccine-associated paralytic polio in India. PMID- 12571733 TI - Nepal's childhood mortality falls by half as vaccinations rise tenfold. PMID- 12571734 TI - Africa's largest measles vaccination campaign could reduce childhood mortality by 20%. PMID- 12571735 TI - Reflectors could reduce road deaths ninefold, says Global Forum for Health Research. PMID- 12571737 TI - Risk reduction can add 5-10 years to healthy life expectancy. PMID- 12571738 TI - WHO and UNICEF focus on eradicating polio in West Africa. PMID- 12571739 TI - Agencies sound the alarm for new strain of meningitis in Africa. PMID- 12571741 TI - Where do we go from here? PMID- 12571742 TI - A new Dual-hemostat technique to facilitate the closure of small laparoscopic trocar incisions. AB - BACKGROUND: The introduction of any surgical procedure brings with it new technical challenges. The use of laparoscopy offers patients a reduction in pain and better cosmesis. However, over time, new technical challenges associated with laparoscopic surgery are becoming more evident. In particular, the closure of the small trocar site fascial incisions is often awkward and time consuming. The aim of this article is to introduce a closing procedure that facilitates the safe approximation of the abdominal fascia in these wounds. METHOD: One hemostat is used to visualize the fascia of one side of the wound and separate the fascia from the underlying abdominal viscera. Another hemostat retracts the opposite side of the wound. The suture needle is passed through the fascia and between the open tips of the hemostat. The tips of the second hemostat grasp the driven needle and carry it out of the wound. CONCLUSION: The use of two hemostats to close these wounds not only facilitates the closure but is also probably safer than the standard technique. PMID- 12571743 TI - Port placement for laparoscopic appendectomy with the best cosmesis and ergonomics. PMID- 12571744 TI - Is development of bronchial metastasis following laparoscopic right hemicolectomy for colon cancer related to pneumoperitoneum? PMID- 12571745 TI - Comment on gastrointestinal quality of life in patients with symptomatic or asymptomatic cholelithiasis before and after laparoscopic cholecystectomy. PMID- 12571747 TI - Comment on bidirectional endoscopy in patients with fecal occult blood. PMID- 12571750 TI - Molecular mechanisms of urea transport. AB - Physiologic data provided evidence for specific urea transporter proteins in red blood cells and kidney inner medulla. During the past decade, molecular approaches resulted in the cloning of several urea transporter cDNA isoforms derived from two gene families: UT-A and UT-B. Polyclonal antibodies were generated to the cloned urea transporter proteins, and their use in integrative animal studies resulted in several novel findings, including: (1) UT-B is the Kidd blood group antigen; (2) UT-B is also expressed in many non-renal tissues and endothelial cells; (3) vasopressin increases UT-A1 phosphorylation in rat inner medullary collecting duct; (4) the surprising finding that UT-A1 protein abundance and urea transport are increased in the inner medulla during conditions in which urine concentrating ability is reduced; and (5) UT-A protein abundance is increased in uremia in both liver and heart. This review will summarize the knowledge gained from studying molecular mechanisms of urea transport and from integrative studies into urea transporter protein regulation. PMID- 12571751 TI - Gating of Cl- currents in protoplasts from the marine alga Valonia utricularis depends on the transmembrane Cl- gradient and is affected by enzymatic cell wall degradation. AB - The electrical properties of protoplasts of the turgor pressure-regulating giant marine alga Valonia utricularis were investigated by using the patch-clamp technique. In the whole-cell configuration, large inward currents were elicited by negative-going voltage pulses. The time-dependent component was predominantly carried by Cl-, as revealed by 'tail current' analysis. When experiments were performed on protoplasts directly after mechanical release from the 'mother cell', small outward currents were additionally observed at membrane voltages more positive than ECl-. These outward currents disappeared to a large extent after treatment of the protoplasts with a mixture of cell wall-degrading enzymes. Plots of the chord conductance versus the clamped membrane voltage revealed that enzymatic treatment affected the gating properties. By fitting Boltzmann distributions to the data, a midpoint potential of + 5 +/- 5 mV (n = 7) was obtained in symmetrical Cl- solutions for mechanically released protoplasts. In contrast, protoplasts treated additionally with enzymes exhibited a midpoint potential of -13 +/- 5 mV (n = 8). By varying the external and internal Cl- concentration, gating was also shown to depend on the Cl- gradient across the plasmalemma both in enzymatically treated and untreated protoplasts. Plotting of the midpoint potential against the Nernst potential of Cl- rendered a slope less than 1 (0.70 and 0.64, respectively) indicating that gating did not strictly depend on the electrochemical Cl- gradient. The voltage- and Cl--dependence as well as inhibition experiments with 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) suggested that the Cl- conductance of the membrane is dominated by the Valonia Anion Channel 1 (VAC1) described by Heidecker, M., Wegner, L.H., Zimmermann, U. 1999: A patch-clamp study of ion channels in proto-plasts prepared from the marine alga Valonia utricularis. J. Membrane Biol. 172:235-247. The relevance of the findings for membrane potential control and turgor regulation in V. utricularis as well as the general implications of the data for electrophysiological work on protoplasts (that are usually obtained by enzymatic digestion of plant tissue) are discussed. PMID- 12571752 TI - Ca2+ dynamics during membrane excitation of green alga Chara: model simulations and experimental data. AB - Kinetic investigations of stimulus response coupling in the green alga Chara have revealed that an intermediate second messenger is formed in the process of membrane excitation. This second messenger links electrical stimulation to the mobilization of Ca2+ from internal stores. In the present work, the experimentally based kinetic model, which describes the stimulus-dependent production of the second messenger and Ca2+ mobilization, is combined with a model for inositol 1,4,5-trisphosphate (IP3)-and Ca2+-sensitive gating of a Ca2+ release channel in endomembranes of animal cells. The combination of models allows a good simulation of experimental data, including the all-or-none-type dependence of the Ca2+ response on stimulus duration and complex phase locking phenomena for the dependence of the Ca2+ response on stimulation frequency. The model offers a molecular explanation for the refractory phenomenon in Chara, assigning it to the life time of an inactive state of the Ca2+-release channel. The model furthermore explains the steep dependence of excitation on strength/duration of electrical stimulation as a consequence of an interplay of the dynamical variables in the model. PMID- 12571754 TI - Response to ADP-ribose by activation of TRPM2 in the CRI-G1 insulinoma cell line. AB - The response to intracellular ADP-ribose in the rat CRI-G1 insulinoma cell line was studied using a patch-clamp method. Dialysis of ADP-ribose into cells induced a response in a dose-dependent manner. The reversal potentials in various solutions showed that the ADP-ribose-gated channel was a Ca2+-permeable nonselective cation channel. In inside-out recordings, ADP-ribose and b-NAD induced responses in the same patch. The single-channel current-voltage relationships for ADP-ribose- and b-NAD-induced responses were almost identical, indicating that ADP-ribose and b-NAD activated the same channel. The physiological properties of the ADP-ribose-gated channel are similar to those we reported previously for the cloned transient receptor potential channel TRPM2. Moreover, RT-PCR analysis showed that TRPM2 was abundantly expressed in CRI-G1 cells, suggesting that the ADP-ribose-gated channel represents the native TRPM2 channel in CRI-G1 cells. These results suggest that ADP-ribose can be an endogenous modulator of Ca2+ influx through the TRPM2 channel into CRI-G1 cells. PMID- 12571753 TI - Bending the primary cilium opens Ca2+-sensitive intermediate-conductance K+ channels in MDCK cells. AB - Increasing tubular fluid flow rate has previously been shown to induce K+ secretion in mammalian cortical collecting duct. The mechanism responsible was examined in the present study using MDCK cells as a model. The change in membrane potential difference (EM) of MDCK cells was measured with a fluorescent voltage sensitive dye, DiBAC4(3), when the cell's primary cilium was continuously bent with a micropipette or by the flow of perfusate. Bending the cilium produced a hyperpolarization of the membrane that lagged behind the increase in intracellular Ca2+ concentration by an average of 36 seconds. Gd3+, an inhibitor of the flow-induced Ca2+ increase, prevented the hyperpolarization. Blocking K+ channels with Ba2+ reduced the flow-induced hyperpolarization, implying that it resulted from activation of Ca2+-sensitive K+ channels. Further studies demonstrated that the hyperpolarization was diminished by the blocker of Ca2+ activated K+ channels, charybdotoxin, whereas iberiotoxin or apamin had no effect, results consistent with the activation of intermediate-conductance Ca2+ sensitive K+ channels. RT-PCR analysis and sequencing confirmed the presence of intermediate-conductance K+ channels in MDCK cells. We conclude that the increase in intracellular Ca2+ associated with bending of the primary cilium is the cause of the hyperpolarization and increased K+ conductance in MDCK cells. PMID- 12571755 TI - Thyroid hormones affect the membrane dipolar organization. Is it a general event in their non-genomic action? AB - The surface balance technique was employed to study the interactions of 3,5,3',5' tetraiodo L-thyronine, 3,5,3' triiodo L-thyronine, and 3,5-diiodothyronine with monomolecular phospholipid monolayers spread at the air-water interface. With this technique the insertion of thyroid hormones into egg yolk phosphatidylcholine was investigated. An increase of surface pressure and a substantial decrement in surface potential were observed after the injection of these hormones beneath a phospholipid monolayer. The negative dipole contribution upon hormone interaction opposes the well-known positive contribution of phospholipids. These effects correlated with iodo content of the thyroid molecule analogues 3,5,3',5' tetraiodo L-thyronine >3,5,3' triiodo L-thyronine >3,5 diiodothyronine. To our knowledge, these observations suggest a new and surprising effect of thyroid hormones on the regulation of transmembrane dipolar organization. PMID- 12571756 TI - Role of GPI-anchored enzyme in liposome detergent-resistance. AB - In this work, we investigated the role of a glycosylphosphatidylinositol (GPI) anchored protein, the alkaline phosphatase, on the solubilization of detergent resistant liposomes. In vivo, GPI-anchored proteins are clustered into sphingolipid- and cholesterol-rich membrane domains and this peculiar composition provides cold-detergent-insolubility. To better understand the mechanisms involved in the clustering of these subdomain components, we built a model, namely sphingolipid- and cholesterol-rich liposomes. We show the cold-Triton X 100 resistance of liposomes before and after insertion of GPI-anchored enzyme. When the amount of incorporated enzyme varied, significant changes in membrane stability occurred. Low protein contents into liposomes increased detergent insolubility, whereas high amounts decreased it. Furthermore, significant differences in the detergent-resistance of each lipid were exhibited between liposomes and proteoliposomes. Thus, the enzyme insertion led to a dramatic decrease of cholesterol solubilization, in line with the existence of cholesterol/GPI interactions. Effect of temperature on detergent resistance was also investigated. Liposome solubilization increased with temperature up to a threshold value of 40/45 degrees C. This was also the temperature at which a phase transition of liposome membrane occurred, as evidenced by Laurdan fluorescence. Although the GPI-anchored enzyme insertion modified membrane stability, no change was observed on phase transition. Our work highlights the importance of GPI-anchored proteins in the structure of sphingolipid- and cholesterol-rich membrane domains, in the detergent-insolubility of these peculiar domains, as well as in interaction of GPI proteins with cholesterol. PMID- 12571757 TI - Latin as the language of medical terminology: some remarks on its role and prospects. AB - The present paper offers an up-to-date view of the status of Latin as the language of medicine, namely in its terminological component. It is concerned in greater detail with the three basic terminological vocabularies in which a doctor cannot so far manage without its knowledge. In this sense a primary rank is occupied by anatomical nomenclature whose international version remains Latin in the full extent. A more varied picture is presented by the clinical disciplines where, apart from Latin terms, expressions of ancient provenance have been applied in a large measure in the form of ethnic languages. At the same time, particularly in view of the needs of computerisation, repeated attempts have appeared to support English, which has the greatest chance of becoming a new language in the particular region of clinical medicine. In pharmaceutical terminology Latin has, for the time being, remained a functioning means of international communication, guaranteed by the European Pharmacopoeia (1996) and by the corpus of International Non-proprietary Names (1992, 1996), even though in the future an ever stronger competition of national languages should be taken into account. PMID- 12571758 TI - Update on cryptogenic organising pneumonia (idiopathic bronchiolitis obliterans organising pneumonia). AB - Organising pneumonia, defined by intraalveolar buds of connective tissue, may be a disorder secondary to a determined cause (infectious agents, drugs) or occurring in a specific context (as the connective tissue disorders). It may also be a cryptogenic interstitial pneumonia with characteristic clinical and imaging features and especially an excellent response to corticosteroids. PMID- 12571759 TI - Evaluation of a population-based prevention program against influenza among Swiss elderly people. AB - INTRODUCTION: Influenza is a major cause of morbidity and mortality and occurs in epidemics in the winter. This study is an evaluation of a population-based prevention program against Influenza, implemented during autumn 2000 by the Health Department of the Canton of Vaud. METHODS: A pre-intervention/post intervention design was used. In June 2000 and March 2001, 4007 questionnaires were sent to two different stratified random samples of people aged 65 and over living in the Canton of Vaud, Switzerland. Univariate, bivariate and multivariate analyses were performed. RESULTS: Vaccination coverage among people older than 65 was 58.0% in 1999 (95% CI: 56.2%-59.8%) and 58.4% (95% CI: 56.6%-60.2%) in 2000. A 6.5% significant increase in vaccination coverage was seen in the group of people aged 65 to 69 (p = 0.008). In the pre-intervention survey immunisation rates were 22.6% among people who had not consulted a physician, 59.2% among those who had consulted a physician once, and 73.2% among those who consulted twice or more (p = 0.001). These rates were respectively 30.8%, 58.0% and 75.1% (p = 0.001) in the post-intervention survey. CONCLUSIONS: No global increase in Influenza vaccination coverage in the elderly population could be observed following a community based intervention in a Swiss Canton. However, the enhanced vaccination rates noted in the 65-69 years old group and in people who did not receive medical care are compatible with an effect of the campaign. Further increase in vaccination coverage may be obtained by diversification and repetition of such promotion campaigns. PMID- 12571760 TI - Congenital tuberculosis in a premature infant. AB - Congenital tuberculosis was first suspected in a premature infant with patent ductus arteriosus (PDA), progressive respiratory distress and septic shock, when an enlarged mediastinal lymph node was noted intraoperatively at the time of PDA ligature. Culture revealed Mycobacterium tuberculosis. The asymptomatic mother was subsequently diagnosed with urogenital tuberculosis. PMID- 12571761 TI - News on lung volume reduction surgery. AB - Lung volume reduction surgery (LVRS) is an established therapeutic option for patients with advanced pulmonary emphysema after all conservative measures, including comprehensive pulmonary rehabilitation, have been exhausted. LVRS improves pulmonary function, shortness of breath, exercise capacity and hence quality of life in some 80% of cases for up to four years. Even patients with homogeneous types of pulmonary emphysema improve if those with extremely low FEV1 and/or very low diffusion capacity are excluded. At experienced centres perioperative mortality is less than 2% in appropriately selected patients, and current results suggest that the five-year survival in COPD patients may even be improved by this palliative surgical intervention. In patients under 60 LVRS may serve as a bridging procedure to lung transplantation. Bronchoscopic creation of extraanatomic bronchopulmonary passages--endoscopic LVRS--is a novel approach now under investigation. PMID- 12571762 TI - Quality of life of Do-Not-Resuscitate (DNR) patients: how good are physicians in assessing DNR patients' quality of life? AB - QUESTIONS UNDER STUDY: To assess 1) the impact of quality of life evaluation on the implementation of Do-Not-Resuscitate (DNR) orders by physicians, 2) the accuracy of physicians' estimation of DNR patients' quality of life. METHODS: A 10-month prospective clinical study in a community hospital including 255 DNR patients and 9 physicians in postgraduate training. Outcomes of interest were the influence of quality of life on the DNR decision and the assessment from patients and physicians of five different components related directly or indirectly to quality of life: mental (the Mini Mental State Examination), physical (the Activities of Daily Living) and social (Framingham Disability), degree of pain (visual analogical scale of pain) and of depression (Geriatric Depression Scale). RESULTS: Quality of life intervened in more than 70% of the DNR decisions. However, physicians underestimated quality of life components of DNR patients (Kappa <0.4 for each functionality). Severe depression, social isolation and physical dependence influenced negatively patients' perception of their quality of life (p <0.01). CONCLUSION: Physicians often (71%) rely on the assumed quality of life of their patients in their DNR decision but unfortunately tend to underestimate it. Greater involvement of patients in the DNR decision could improve quality of care. PMID- 12571763 TI - Rofecoxib improves quality of life in patients with hip or knee osteoarthritis. AB - A major goal of osteoarthritis (OA) treatment is pain management to improve function and maximise quality of life. Rofecoxib is a highly selective inhibitor of cyclooxygenase-2 used in symptomatic treatment of inflammation and pain in patients with osteoarthritis of the hip or knee. AIM: The primary aim of this study was to assess the effects of rofecoxib on quality of life in elderly patients with painful osteoarthritis flares of the hip or knee, who were not responsive to or had adverse reactions to previous NSAID therapy. In addition the switch pattern of NSAIDs in these patients was recorded. METHODS: A 3-week prospective open label multicentre study with rofecoxib 25 mg daily in 134 male and female outpatients with painful osteoarthritis flares of the knee or the hip (mean age 69 years, SD + 8). On day 1 the patients were all switched from their previous NSAID to rofecoxib, followed by continuous daily treatment with rofecoxib 25mg daily over 3 weeks. On day 21 the patients discontinued daily treatment with rofecoxib and had the choice between either staying on rofecoxib, switching back to their previous NSAID, trying another NSAID or stopping drug treatment. The impact on quality of life was measured by the difference in SF-12 between day 0 and day 21. Further endpoints included changes in self-reported pain, stiffness and functional ability as measured by the WOMAC index (Western Ontario McMaster Universities Osteoarthritis Index). Correlation studies were performed between the WOMAC pain subscale and quality of life as measured by the SF-12 at baseline and over the course of the study. Patients' report of general health status and overall assessment of pain intensity, as measured by visual analogue scale (VAS), was correlated with physicians' and patients' assessment of the efficacy of rofecoxib treatment. RESULTS: Quality of life improved with rofecoxib: the physical component summary score (SF-12 PCS) was improved by a statistically significant +16.2% (p <0.0001) after 3 weeks, while the mental health component summary score (MCS) was improved by +3.0% (n.s.). Disease specific symptoms measured by the WOMAC questionnaire were significantly improved under rofecoxib after 3 weeks: pain decreased by 29% (p <0.0001) and stiffness by 25% (p <0.0001), while functional ability increased by 24% (p <0.0001). The improvement in SF-12 PCS correlated negatively with the decrease in WOMAC scores (r = -0.54, p <0.0001; r = -0.46, p <0.0001 and r = -0.64, p <0.0001 respectively). General health was significantly improved by +30.5% (or 15.96 mm, p <0.0001) between baseline and day 21, while pain was significantly reduced by 35.2% (or 17.67 mm, p <0.0001) on the VAS scales. At the end of the 3-week study 75% of the patients and 84% of the treating physicians rated the efficacy of rofecoxib from good to excellent. Two weeks after study end the planned telephone survey revealed that 54% of the patients preferred to stay on therapy with rofecoxib, 19% had decided to switch back to their previous NSAID (this observation being most marked for diclofenac, where 38% of initial diclofenac patients had decided to switch back to their initial therapy), 9% had been switched to another NSAID and 7.5% had discontinued treatment. The switch pattern is unknown in the remaining 7.5%. CONCLUSION: Rofecoxib significantly improves quality of life, as measured by the SF-12, in OA patients who were either unresponsive to or presented with adverse reactions to previous NSAID therapy (including celecoxib). In addition, rofecoxib significantly improved pain, stiffness and function, as assessed by the WOMAC questionnaire. PMID- 12571764 TI - Premenstrual dysphoric disorder: current status of treatment. AB - OBJECTIVE: Premenstrual dysphoric disorder (PMDD), also referred to as premenstrual syndrome (PMS), is a recurrent luteal-phase condition involving regular occurrence, prior to the onset of menstrual bleeding, of a cluster of symptoms of sufficient severity to result in the deterioration of interpersonal relationships and normal activity. Several treatment options for PMDD with varying degrees of efficacy have been proposed. The literature is reviewed and treatments of proven efficacy are reported. STUDY DESIGN AND METHODS: A MEDLINE/Cochrane Library search for all studies on PMS and PMDD published between 1983 and 2001 was performed. Only randomised trials were included. RESULTS: Several treatments appear to be effective. Among these are increased physical activity, dietary change, mineral salt supplementation and ovulation inhibitors. The most effective seems to be administration of selective inhibitors of serotonin reuptake (SSRIs). CONCLUSION: Therapy should begin with nonmedicated approaches and pharmacological treatment should only be envisaged if symptoms persist. PMID- 12571766 TI - Advances in a gene therapy-based approach to treat proliferative vitreoretinopathy. PMID- 12571765 TI - [Adapting contact lenses in extraordinary cases: an ophthalmologist's field]. PMID- 12571767 TI - [New advances in aniridia]. PMID- 12571768 TI - [Botulinum toxin as a treatment for strabismus in systemic diseases]. AB - PURPOSE: To demonstrate the effectiveness of botulinum toxin for treating strabismus secondary to systemic diseases. MATERIAL AND METHODS: 141 patients with secondary strabismus were studied. To reduce risks, the treatment with botulinum toxin was made by a direct technique proposed by the authors. The response to the treatment was classified according to the different physiopathogenic groups. RESULTS: We found a positive response in: Central Neurological damage: 71%, Endocrinopathies: 78.6%, Brain infection: 80%, Trauma: 60%, Psychomotor defficiency: 72%, Prematurity: 74%, Myasthenia 75%, Hemathological diseases: 75%. The average of botulinum toxin applications was 1.5 infections. CONCLUSIONS: We show that the effectiveness of botulinum toxin in strabismus secondary to systemic disease is up to 74%. We can offer rehabilitation in all of these cases even during the sometimes long diagnostic period or in patients under treatment. We also suggest our direct technique for the application of botulinum toxin (without electromyography) to avoid risks in this type of patients. We are reporting the use and positive effect of the botulinum toxin chemodenervation in Myasthenia gravis, Human Acquired Immune deficiency Syndrome, and mental defficiency. PMID- 12571769 TI - [Benefit of scanning laser ophthalmoscopy for detecting retinal nerve fiber layer defects in patients with intraocular hypertension]. AB - PURPOSE: To compare the ability to identify RNFL alterations between standard images and SLO images in a group of patients suffering from ocular hypertension. METHODS: We evaluated RNFL in sixty patients with ocular hypertension and with normal perimetry, by using standard photography and SLO. Two different independent observers evaluated the images on two different ocassions. RESULTS: We studied 118 eyes, twelve eyes were rejected because of the low quality of the images. Four cases of diffuse atrophy and five patients with wedge shaped defects were observed using both methods. Statistical analysis showed no differences between both exploration techniques or between observers. CONCLUSIONS: RNFL study with SLO seems to be at least as reliable as standard photography as a method to evaluate RNFL defects. SLO has also proved to be much easier, and faster than traditional methods. PMID- 12571770 TI - [Lacunar folds study in dry eye diagnosis]. AB - PURPOSE: To determine the diagnostic value of lacunar folds study in patients with dry eye syndrome. METHODS: We present a prospective study in 185 patients (91 males and 94 females). In all patients the abduction degree was measured after instilling a fluorescein drop, which makes lacunar folds disappear. We divided patients into two groups, one of them with dry eye symptoms and the other without them. The symptomatic group was divided into four other subgroups, according to the severity of symptoms. RESULTS: We found significant differences in the early disappearing of lacunar folds in dry eye patients compared with the control group. The folds disappeared quicker when the symptoms were more severe. The disappearance of folds is significantly associated with the Schirmer test and with the break up time test (BUT). CONCLUSIONS: Lacunar folds study should be included as a useful and easy test in the diagnosis of dry eye syndrome, nevertheless its disappearance is not exclusive of this disease. PMID- 12571771 TI - [Study of two immunosuppresive strategies after limbal allograft transplantation]. AB - PURPOSE: Comparison of two immunosuppressive strategies after limbal allograft transplantation in an experimental model of ocular surface chemical burn. METHODS: Twenty albino rabbits underwent corneal and limbal chemical burn with NaOH. One week later limbal transplantation was performed. Clinical assessments included measurements of epithelial defect, stromal opacity and neovascularization areas, using a computerized planimetry system 0, 7, 15, 30, 60 and 90 days after the chemical burn. Histologic examination was performed at 3 months, after sacrifice of the animals. Two immunosuppressive strategies were evaluated: topical cyclosporin A on constant dosage (group 1), and a combination of topical cyclosporin A and dexamethasone followed by a tapered dosage of topical cyclosporin A (group 2). RESULTS: No statistical differences were observed between both groups regarding all of the variables 3 months after the chemical burn. Differences in the stromal opacity area were detected on day 7 (43.33 D.E. 4.84 vs 36.67 D.E. 3.88; p<0.01), day 15 (41.68 3 D.E.95 vs 33.6 D.E. 1.99; p<0.01), and day 30 (40.1 D.E. 5.45 vs 32.52 D.E. 7.33; p<0.05). As for the neovascularization area, differences were detected on day 7 (1.92 D.E. 0.48 vs 3.73 D.E. 1.18; p<0.01), day 15 (7.08 D.E. 1.54 vs 4.42 D.E. 1.68; p<0.01), and day 60 (4.9 D.E. 2.58 vs 9.65 D.E. 5.55; p<0.05). CONCLUSIONS: There are no long term differences between both immunosuppressive strategies regarding all the studied variables (Arch Soc Esp Oftalmol 2003; 78: 29-34). PMID- 12571772 TI - [Retinal pigment epithelial atrophic tracks secondary to central serous pigment epitheliopathy]. AB - PURPOSE AND METHODS: We report the cases of three patients with retinal pigment epithelial atrophic tracks secondary to central serous pigment epitheliopathy specially focusing on fluorescein angiographic findings. This entity is considered an atipical manifestation of central serous chorioretinopathy called chronic diffuse retinal pigment epitheliopathy. RESULTS AND CONCLUSIONS: We have studied 69 patients with the diagnose of central serous pigment epiteliopathy in our unit; 10.14% manifested diffuse retinal pigment epitheliopathy and 5.79% showed retinal pigment epithelial atrophic tracks. Both eyes were involved in 100% and all of them were males with a median age of 53.33 years. We can conclude that this entity usually affects adult males and it is frequently bilateral. PMID- 12571773 TI - [Can the use of intraocular silicone oil increase the risk of sympathetic ophthalmia?]. AB - CLINICAL CASES: We report two cases of SO (sympathetic ophthalmia) among 1,456 vitrectomies and 255 silicone oil injections performed since 1990. In both patients intraocular silicone oil was used. Both cases were clinically documented and one pathologically demonstrated. DISCUSSION: No cases were reported among vitrectomies without silicone oil. We found a longer delay between injury and the onset of the disease in comparison to the literature. Pathological findings confirmed a foreign body granulomatous reaction. Chronic inflammation caused by silicone oil might be involved in the development of SO. Larger series are necessary to establish a possible relationship between silicone oil and SO. PMID- 12571774 TI - [Painful ophthalmoplegia secondary to larynx carcinoma]. AB - CASE REPORT: We present a case of metastatic larynx cancer in the cavernous sinus. It presented itself as a right sixth cranial nerve palsy which progressed into a painful ophthalmoplegia with normal CT and RNM in its early stages. DISCUSSION: We must suspect a cavernous sinus invasion in patients with cranial nerve palsies and with a history of malignancy, despite negative radiologic findings. Tumoral invasion of the skull base has been described in pharyngeal neoplasms but it is exceptional in larynx carcinomas. PMID- 12571775 TI - [Adult-onset foveomacular vitelliform dystrophy]. AB - CLINIC CASES: Four cases of adult-onset foveomacular vitelliform dystrophy are shown. In two cases we were able to complete the exploration under optical coherence tomography. One of the patients developed spontaneous disappearance of the subfoveal deposit in her right eye along with an important decrease in her visual acuity. DISCUSSION: Adult-onset foveomacular vitelliform dystrophy is characterized by yellowish round-shaped bilateral, lesions in the subfoveal area. Optical coherence tomography reveal the presence of a slightly elevated lesion wich affects the retinal pigment epithelium and the external layers of neurosensory retina. PMID- 12571776 TI - [Diplopia as the first manifestation of Lyme disease]. AB - CASE REPORT: We report a case of Lyme disease with diplopia as the first manifestation, without systemic symptoms in contrast with other cases of this disease. The serodiagnosis was confirmed by ELISA analysis and evaluation of cerebrospinal fluid (CSF) antibodies. Symptoms readily vanished after the introduction of antibiotherapy. DISCUSSION: Lyme disease is an infectious multi systemic disorder caused by Borrelia burgdorferi. Neuro-ophthalmological manifestations occur during the second stage of the illness in extremely variable clinical forms. PMID- 12571777 TI - [Conjunctival infection due to penicillium SP]. AB - CLINICAL CASE: We present a case of conjunctivitis in a diabetic patient in whom not improvement was observed after empiric treatment, and finally developed a conjunctival granuloma. Culture of the conjunctival exudate and histological examination of the conjunctival biopsy confirmed the infection by Penicillium sp. The antifungical treatment (local and systemic) improved signs and symtoms gradually. DISCUSSION: Penicillium sp is an infrequent cause of conjunctival infection. This diagnosis should be considered when no response after usual therapy to conjunctivitis is obtained and should be confirmed by identification of the fungus in the lesions biopsed and cultured. Succesful management of ocular penicilliosis is achieved with both local and systemic antifungal treatment. PMID- 12571778 TI - [A century of anaphylaxis and allergy]. PMID- 12571779 TI - [Op art]. PMID- 12571781 TI - Adult stroke and perinatal brain damage: like grandparent, like grandchild? AB - Biomarkers of inflammation are found in the circulation of adults who have had a stroke. Although these biomarkers may, in part, be indicators of damage, some appear to contribute to damage. Similar biomarkers are found in newborns with cerebral white matter damage or at risk of cerebral palsy. Can we learn about the pathogenesis of neonatal white matter damage from what has been learned about the inflammatory correlates of adult stroke? We discuss relevant findings about systemic inflammatory markers in adult stroke and relate this information to our current understanding of cerebral white matter damage in newborns, especially those born at an extremely low gestational age. We also describe desirable characteristics of future studies of perinatal brain damage that involve measurements of systemic biomarkers. PMID- 12571782 TI - Paroxysmal kinesigenic dyskinesia and generalized seizures: clinical and genetic analysis in a Spanish pedigree. AB - Familial paroxysmal kinesigenic dyskinesia (PKD) is a rare disorder featuring brief, dystonic or choreoathetotic attacks, typically triggered by sudden movements. Symptoms usually start in mid-childhood, although in several pedigrees infantile convulsions have been reported as the presenting sign. Previous linkage studies have identified two PKD loci on 16 p12.1-q21. We report here the clinical features of a Spanish kindred with autosomal dominant PKD, in which haplotype data are compatible with linkage to the pericentromeric region of chromosome 16 and exclude linkage to the locus for Paroxysmal Non Kinesigenic Dyskinesia (PNKD) on chromosome 2 q35. In this family, the conservative candidate region for the disease lies between markers D16S3145 and GATA140E03 on 16 p12.1-q21 and partially overlaps with both the Paroxysmal Kinesigenic Dyskinesia - Infantile Convulsions (PKD-IC) critical interval and the Episodic Kinesigenic Dyskinesia 2 (EKD2) locus. Unusual findings in our pedigree were early infantile onset of the dyskinesias in one patient and generalized seizures as adults in two, adding to previous observations of phenotypic overlap between epileptic and non-epileptic paroxysmal disorders. Further clinical and genetic studies are needed to elucidate whether PKD and PKD-IC are allelic disorders with age-dependent phenotypic expression. PMID- 12571783 TI - Eating epilepsy characterized by periodic spasms. AB - Seizures induced by eating are generally considered rare. Eating epilepsy is a kind of reflex epilepsy. We report two patients aged 11 and 16, with seizures provoked by eating. The eating-provoked seizures in both patients were a series of tonic spasms. Both ictal EEGs showed a periodic pattern characterized by diffuse high voltage slow wave. These findings were consistent with periodic spasms described by Gobbi et al. The ECD were widely distributed in both patients, although that of Patient 1 was partially clustered in the right frontal region. This is the first report of patients with periodic spasms induced by eating. PMID- 12571784 TI - Oxcarbazepine-induced hyponatremia and the regulation of serum sodium after replacing carbamazepine with oxcarbazepine in children. AB - While severe hyponatremia is reported to be more frequent in adults treated with oxcarbazepine (OXC) than with carbamazepine (CBZ), there is not sufficient data about the incidence of hyponatremia in childhood during treatment with OXC. We evaluated changes in serum electrolyte balance in 75 children with epilepsy before and during treatment with OXC and after replacing carbamazepine (CBZ) therapy with OXC therapy. All patients had normal sodium serum levels at the onset of OXC. During treatment with OXC we found hyponatremia (Na +< 135 mmol/l) without clinical symptoms in 26.6 % of the children (n = 20), sodium levels below 125 mmol/l were observed in 2 children (2.6 %). Clinically relevant hyponatremia occurred in one girl only (1.3 %). In a subgroup of 27 children, in whom CBZ was directly replaced with OXC, hyponatremia without symptoms was found in one child under CBZ (3.7 %) and in six children under OXC (22.2 %). Dosage of OXC, serum levels of the active metabolite of OXC, antiepileptic comedication or patients' age and gender were of no predictive value for the development of hyponatremia. Electrolytes should be measured before establishing OXC and if clinically relevant side effects occur. PMID- 12571785 TI - Psychomotor retardation, spastic paraplegia, cerebellar ataxia and dyskinesia associated with low 5-methyltetrahydrofolate in cerebrospinal fluid: a novel neurometabolic condition responding to folinic acid substitution. AB - INTRODUCTION: Normal brain development and function depend on the active transport of folates across the blood-brain barrier. The folate receptor-1 (FR 1) protein is localized at the basolateral surface of the choroid plexus, which is characterized by a high binding affinity for circulating 5-methyltetrahydrofolate (5-MTHF). PATIENTS AND METHODS: We report on the clinical and metabolic findings among five children with normal neurodevelopmental progress during the first four to six months followed by the acquisition of a neurological condition which includes marked irritability, decelerating head growth, psychomotor retardation, cerebellar ataxia, dyskinesias (choreoathetosis, ballism), pyramidal signs in the lower limbs and occasional seizures. After the age of six years the two oldest patients also manifested a central visual disorder. Known disorders have been ruled out by extensive investigations. Cerebrospinal fluid (CSF) analysis included determination of biogenic monoamines, pterins and 5-MTHF. RESULTS: Despite normal folate levels in serum and red blood cells with normal homocysteine, analysis of CSF revealed a decline towards very low values for 5 methyltetrahydrofolate (5-MTHF), which suggested disturbed transport of folates across the blood-brain barrier. Genetic analysis of the FR 1 gene revealed normal coding sequences. Oral treatment with doses of the stable compound folinic acid (0.5-1 mg/kg/day Leucovorin(R)) resulted in clinical amelioration and normalization of 5-MTHF values in CSF. CONCLUSION: Our findings identified a new condition manifesting after the age of 6 months which was accompanied by low 5 MTHF in cerebrospinal fluid and responded to oral supplements with folinic acid. However, the cause of disturbed folate transfer across the blood-brain barrier remains unknown. PMID- 12571786 TI - Microlissencephaly in microcephalic osteodysplastic primordial dwarfism: a case report and review of the literature. AB - Microcephalic osteodysplastic primordial dwarfism (MOPD) is defined as a syndrome presenting with intrauterine and postnatal growth retardation, typical facial appearance, skeletal dysplasia and brain abnormalities. Autosomal-recessive inheritance is suspected. Sharing clinical manifestations, the former type III has been accepted to be the same entity as type I. We present the case of a male infant with MOPD I and micrencephaly with simplified gyral pattern to a degree defining it as microlissencephaly (MLIS). The brain abnormalities in MOPD I have not yet been classified. Reviewing the literature, we conclude that microlissencephaly appears to be the distinct developmental brain abnormality in MOPD I. Conversely, osteodysplastic changes have to be taken into consideration in the differential diagnosis of microlissencephaly. In addition, our patient suffered from acute lymphatic leukaemia which has not previously been described in association with MOPD I. PMID- 12571787 TI - Spinal muscular atrophy with progressive myoclonic epilepsy: report of new cases and review of the literature. AB - Spinal muscular atrophy (SMA) is a clinically and genetically heterogeneous disease characterised by loss of motor function and muscle atrophy due to anterior horn cell degeneration. The most common variant is chromosome 5-linked proximal SMA, ranging in severity from congenital onset and infantile death to onset in adult life. Genetically separate variants with different distribution of weakness and/or additional features such as central nervous system involvement have been described. A rare variant with associated myoclonic epilepsy and lower motor neuron disease had been previously described in three families before the SMN gene, responsible for the common form of SMA, was isolated. We report four patients from two additional families affected by a syndrome characterised by severe and progressive myoclonic epilepsy and proximal weakness, tremor and lower motor neuron disease proven by electrophysiologic and muscle biopsy findings. Extensive metabolic investigations were normal and genetic analysis excluded the SMN gene. This study confirms that the association of myoclonic epilepsy and motor neuron disease represents a separate clinical and genetic entity from chromosome 5-linked SMA, the primary defect of which remains unknown. PMID- 12571788 TI - Refractory photosensitive epilepsy associated with a complex rearrangement of chromosome 2. AB - We describe the relevant clinical and therapeutic parameters in a single patient with a complex chromosome 2 abnormality presenting with refractory myoclonic photosensitive epilepsy. FISH technology using yeast artificial chromosomes (YACs) was employed to determine breakage points, microdeletions and inversions on the affected chromosome. In this patient with refractory photosensitive epilepsy, 12 breakpoints and one small inversion were identified on the abnormal chromosome 2. Our data can be used in further genetic studies on the exact location and identification of photosensitivity genes. PMID- 12571789 TI - Rett females: patterns of characteristic side-asymmetric neuroimpairments at long term follow-up. AB - From a follow-up series of 24 classical Rett syndrome females (13-59 years of age), side asymmetries of neuroimpairments were systematically investigated. A characteristic right-sided dominance in terms of dysfunction and neurology was revealed. Differences in the perinatal/early postnatal development of finer brain net structures between brain halves are suggested as a possible explanation. PMID- 12571790 TI - Successful management of drooling with botulinum toxin A in neurologically disabled children. AB - OBJECTIVES: The present study investigates the effect of topical injections of botulinum toxin A into the cephalic salivary glands of children with chronic hypersalivation due to neurodegenerative diseases. METHODS: Five children with hypersalivation due to severe neurological diseases received, under ultrasound guidance, a total of 50-65 units of botulinum toxin A (Botox) into the parotid and submandibular glands on both sides. All injections were tolerated without local anaesthesia. Before and 1, 2, 4, 8 and 12 weeks after toxin injection, salivary flow rates and the concentrations of total protein, alpha-amylase, acid phosphatase, kallikrein, and immunoglobulin A were measured in the secretions; simultaneously, the patients were clinically examined with regard to severity of symptoms, and their salivary glands were subjected to ultrasound study. RESULTS: A distinct improvement of symptoms within the first 2 weeks following toxin administration were reported by the parents. Sialometry revealed considerably reduced flow rates but sialochemistry showed an increase of amylase activity. Ultrasound examination did not reveal any changes of the salivary parenchyma, and side-effects were absent. CONCLUSION: Treatment of drooling by topical injection of botulinum toxin A into the salivary glands is a reliable and also side-effect free therapeutic option for children with neurological disorders. All children involved in our study experienced a distinct improvement of their quality of life. PMID- 12571791 TI - Diagnostic difficulties in childhood bilateral thalamic astrocytomas. AB - We report on two children with bilateral thalamic astrocytomas. The first patient developed psychomotor regression at the age of 20 months followed by rapidly progressive ataxia, intention tremor, slurred speech, and bouts of drowsiness. Magnetic resonance imaging (MRI) of the brain showed swelling and high signal intensity in both thalami accompanied by supratentorial hydrocephalus. The second patient presented with progressive cerebellar ataxia, headache, and vomiting at the age of 11 years. MRI of the brain revealed symmetrical, hyperintense and sharply delineated swelling of both thalami. Additional lesions were seen in the cerebellum and the right temporal lobe. In both cases proton magnetic resonance spectroscopy (MRS) of the lesions showed a striking decrease of the neuronal marker N-acetylaspartate, an increase of choline-containing compounds, and a minimal lactate peak. Stereotactic biopsies from the thalamus of the first patient and from a cerebellar lesion of the second patient finally revealed glial tumors, namely a diffuse astrocytoma of World Health Organization (WHO) grade II in the first patient and an anaplastic astrocytoma of WHO grade III in the second patient. We conclude that the clinical manifestations and MRI patterns of bilateral thalamic astrocytomas are very similar to those of encephalitis and neurometabolic disorders and should therefore be included in the differential diagnosis of these encephalopathies. PMID- 12571792 TI - [Severe hypoglycemia in diabetics with impaired renal function]. AB - BACKGROUND AND OBJECTIVE: Reduction of renal function in patients with type 1 or type 2 diabetes is associated with a clearly increased risk of hypoglycemia. Main causes are an altered pharmacokinetics of insulin and oral antidiabetics and/or impaired renal glucose production. A knowledge of renal function is, therefore, essential for preventing hypoglycemia caused by antidiabetic treatment. But serum creatinine, most commonly used in general practice, is an imprecise indication of renal function. This investigation assessed the significance of a false estimation of renal function as a partial cause of severe hypoglycemia. PATIENTS AND METHODS: The study group consisted of 35 diabetics (21 females, 14 males; average age 61 years) who had been hospitalized because of an episode of severe hypoglycemia accompanied by loss of consciousness. Renal function was measured by serum creatinine and by creatinine clearance as calculated by the formula of Cockcroft and Gault. Also taken into account were HBA1c the antidiabetic treatment before and after discharge, and any additional medication. RESULTS: Impaired renal function was established by the serum creatinine level in 9 patients and by calculated creatine clearance in 24. Compared with patients with normal renal function, those with renal failure were older (74.3 vs. 32.8 years), had more rarely undergone intensive insulin treatment (5 of 24 vs. 9 of 11) and had more commonly received ACE inhibitors (10/24 vs. 1/11). The insulin dosage at discharge had been reduced in 16 off 22 insulin-dependent patients in renal failure, and long-acting sulfonylurea preparation were discontinued or changed to gliquidone in the others. CONCLUSION: This investigation indicates that false estimation of renal function from the level of serum creatinine is an important partial cause of hypoglycemia requiring treatment, especially in elderly persons with reduced muscle mass in whom renal function should be determined by calculating or measuring creatinine clearance. PMID- 12571793 TI - [Hypoglycemia associated with the production of insulin-like growth factor (IGF) II by a hemangiopericytoma]. AB - HISTORY AND ADMISSION FINDINGS: A seventy-seven year-old woman with an unclear tumor of the liver suffered from recurrent hypoglycemia and was therefore admitted to our hospital. As diabetes mellitus, hyperinsulinism and reactive forms of hypoglycemia could be excluded, the presumptive diagnosis was non-islet cell tumor hypoglycemia (NICTH). INVESTIGATIONS: Postprandial glucose levels were normal. Fasting glucose levels were 30 - 50 mg/dl. Plasma insulin-like growth factor (IGF)-I was below the normal range, IGF-II was not elevated, although 34 % of plasma IGF-II was present as "big"-IGF-II. IGF-binding protein (IGFBP)-2 was extremely elevated, whereas IGFBP-3 was within the normal range. Histological examinations of the tumor revealed a hemangiopericytoma of the liver. TREATMENT AND COURSE: : After a 2-month treatment with steroids and an experimental antiangiogenetic therapy, the glucose metabolism became stable. The tumour did not grow. Simultaneously, plasma IGF-II and "big"-IGF-II remained constant and plasma IGF-I level improved slightly. IGFBP-2, which is presumable produced by the tumor, increased, IGFBP-3 fell below the normal range. CONCLUSION: NICTH is a rare but important differential diagnosis of recurrent hypoglycemia. The tumor derived IGF-II has a higher than normal molecular weight ("big"-IGF-II) and shows different interactions with binding proteins, thus resulting in an increased bioavailability. An increased glucose uptake in different tissues as well as inhibition of hepatic gluconeogenesis and lipolysis lead to severe hypoglycemia. If surgical therapy of the tumor is not possible, symptomatic treatment with steroids may represent an effective alternative to control severe hypoglycemia. PMID- 12571794 TI - [Febrile respiratory infection and erythema in a 25-year-old man]. AB - HISTORY AND CLINICAL FINDINGS: A 26-year-old man fell acutely ill with fever (39 degrees C), rigor, dry cough, headache and pain in the neck and limbs. Ambulant treatment with amoxycillin, later cefuroximaxetil, worsened his general condition. At admission to hospital he had, over his trunk and limbs, cocarde like, erythematous efflorescences up to 2 cm in diameter with central elevations and blisters. Additional acute bronchitis, pharyngitis, stomatitis, tonsillitis, conjunctivitis and urethritis indicated extensive mucosal involvement. INVESTIGATIONS: Significant findings were: 9900/microl WBC, ESR 57 mm at one hour, C-reactive protein 160 mg/l, capillary pO2 6.4 kPa, pCO2 4.2 kPa. Mycoplasma serology (days 1, 8, 15) took a typical course. Chest radiogram showed an interstitial infiltrate on day 5. DIAGNOSIS: Respiratory Mycoplasma infection with interstitial pneumonia and exudative erythema multiforme. TREATMENT AND COURSE: Because infection with an atypical pathogen was suspected, antibiotic treatment was changed to erythromycin and, at the suggestion of the dermatologist and ophthalmologist, local treatment (erythromycin, dexamethasone and dexpanthenol) was begun. Mycoplasma serology was first positive on day 3. The fever had disappeared on day 6 and the rash had regressed. Blood gases and blood picture were normal by day 8. The patient was discharged on day 18, after marked improvement of the skin and mucosal changes. CONCLUSION: The occurrence of a rare dermatological complication of a relatively common respiratory Mycoplasma infection can, when the erythema precedes positive Mycoplasma serology, aid in the differential diagnosis. PMID- 12571795 TI - [Acute coronary syndromes without ST-elevation -- case report]. PMID- 12571796 TI - [Acute coronary syndromes without ST-elevation -- diagnosis]. PMID- 12571797 TI - [Acute coronary syndromes without ST-elevation -- treatment]. PMID- 12571801 TI - [Fixed time of work contracts of physicians during continuing education. Decision of the Federal Labor Court, 8/14/2002-7 AZR]. PMID- 12571800 TI - [Impact of GP IIb/IIIa antagonists in interventional cardiology]. PMID- 12571802 TI - Mutations in two nonhomologous genes in a head-to-head configuration cause Ellis van Creveld syndrome. AB - Ellis-van Creveld syndrome (EvC) is an autosomal recessive skeletal dysplasia. Elsewhere, we described mutations in EVC in patients with this condition (Ruiz Perez et al. 2000). We now report that mutations in EVC2 also cause EvC. These two genes lie in a head-to-head configuration that is conserved from fish to man. Affected individuals with mutations in EVC and EVC2 have the typical spectrum of features and are phenotypically indistinguishable. PMID- 12571803 TI - The pedigree rate of sequence divergence in the human mitochondrial genome: there is a difference between phylogenetic and pedigree rates. AB - We have extended our previous analysis of the pedigree rate of control-region divergence in the human mitochondrial genome. One new germline mutation in the mitochondrial DNA (mtDNA) control region was detected among 185 transmission events (generations) from five Leber hereditary optic neuropathy (LHON) pedigrees. Pooling the LHON pedigree analyses yields a control-region divergence rate of 1.0 mutation/bp/10(6) years (Myr). When the results from eight published studies that used a similar approach were pooled with the LHON pedigree studies, totaling >2,600 transmission events, a pedigree divergence rate of 0.95 mutations/bp/Myr for the control region was obtained with a 99.5% confidence interval of 0.53-1.57. Taken together, the cumulative results support the original conclusion that the pedigree divergence rate for the control region is approximately 10-fold higher than that obtained with phylogenetic analyses. There is no evidence that any one factor explains this discrepancy, and the possible roles of mutational hotspots (rate heterogeneity), selection, and random genetic drift and the limitations of phylogenetic approaches to deal with high levels of homoplasy are discussed. In addition, we have extended our pedigree analysis of divergence in the mtDNA coding region. Finally, divergence of complete mtDNA sequences was analyzed in two tissues, white blood cells and skeletal muscle, from each of 17 individuals. In three of these individuals, there were four instances in which an mtDNA mutation was found in one tissue but not in the other. These results are discussed in terms of the occurrence of somatic mtDNA mutations. PMID- 12571805 TI - Molecular, biochemical, and cellular pharmacology of pemetrexed. AB - Pemetrexed is a new-generation antifolate that in its higher polyglutamyl forms is a potent, direct inhibitor of thymidylate synthase and, to a lesser extent, glycinamide ribonucleotide transformylase. Activity of the drug may be partially preserved under conditions in which cells are highly resistant to other thymidylate synthase inhibitors, possibly because of premetrexed's secondary inhibitory effects on purine synthesis. Pemetrexed inhibition of dihydrofolate reductase is not of pharmacologic importance. Pemetrexed has high affinity for the reduced folate carrier and folate receptor and is among the most potent substrates for folylpolyglutamate synthetase. These properties result in rapid accumulation of the free drug in cells with the rapid formation of high levels of the active polyglutamyl congeners. Pemetrexed activity is modulated by natural folates within cells that compete for polyglutamation at the level of folylpolyglutamate synthetase. Cells resistant to methotrexate because of impaired transport via the reduced folate carrier may retain partial sensitivity to pemetrexed. This is due to concurrent diminished transport of physiologic reduced folates and contraction of the cellular folate pool, thereby relaxing the usual level of suppression of pemetrexed polyglutamation. The risk of pemetrexed toxicity is increased when cellular folates are suboptimal. This is best monitored by assessment of blood homocysteine levels, and can be diminished by the coadministration of folic acid. PMID- 12571806 TI - Pemetrexed: single-agent and combination phase I study overview. AB - Pemetrexed is a novel folic acid antimetabolite that exerts its activity by the inhibition of multiple enzyme targets. It has been evaluated in a series of phase I clinical trials that explored different administration schedules. The schedule that has been carried forward into phase II trials involves the administration of pemetrexed as a 10-minute intravenous infusion every 21 days with no standard pretreatment with folic acid or vitamin B(12). When given in this manner, the dose-limiting toxicities were neutropenia and thrombocytopenia. Other toxicities included mucositis, rash, and fatigue. The recommended phase II dose was 600 mg/m(2) administered intravenous every 21 days, although this was subsequently modified to 500 mg/m(2) when several early patients experienced toxicities requiring dose reduction in phase II studies. On the basis of preclinical studies suggesting additive or synergistic effects, pemetrexed has also been evaluated in phase I studies in combination with several other agents including cisplatin, carboplatin, oxaliplatin, irinotecan, the taxanes, and anthracyclines. Encouraging anticancer activity has been observed in many of these studies. PMID- 12571807 TI - Pemetrexed safety and dosing strategy. AB - Pemetrexed is a novel antifolate/antimetabolite that inhibits several folate dependent enzymes, including thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide transformylase. As a class, antifolates have been associated with sporadic severe myelosuppression with gastrointestinal toxicity. Although infrequent, a combination of such toxicities carries a high risk of potentially life-threatening complications. Severe toxicity from pemetrexed-based therapy has become more predictable using the vitamin deficiency marker homocysteine and, to a lesser extent, methylmalonic acid. Evidence now suggests that reducing total plasma homocysteine levels by supplementation with folic acid and vitamin B(12) leads to a better safety profile for pemetrexed, while not adversely affecting its efficacy. PMID- 12571808 TI - Preclinical and clinical studies with combinations of pemetrexed and gemcitabine. AB - The novel antimetabolite pemetrexed inhibits the folate-dependent enzymes thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyltransferase. This agent is broadly active in a wide variety of solid tumors, including non-small cell lung, breast, bladder, head and neck, and ovarian cancers, as well as mesothelioma. Gemcitabine is a pyrimidine nucleoside antimetabolite that is approved worldwide for the treatment of pancreatic and non small cell lung cancers, and bladder cancer outside the United States. In addition, gemcitabine is active against a broad range of tumors including breast, ovarian, and other cancers. Preclinical studies have shown cytotoxic synergy when pemetrexed is combined with gemcitabine. Based on these data, a phase I study of this combination was performed that showed striking activity. Phase II studies of this combination are being performed in breast and non-small cell lung cancer. In addition, a phase III study in pancreatic cancer is ongoing. PMID- 12571809 TI - Emerging insights into the biology and therapy of malignant mesothelioma. AB - Malignant mesothelioma is an aggressive malignancy that may be caused by environmental carcinogens (asbestos and erionite), viruses (SV40), and genetic predisposition. Pleural malignant mesotheliomas are far more common than the peritoneal variants. Diagnosis relies on radiographic studies as well as histology and molecular biologic analyses. The prognostic scoring systems of the Cancer and Leukemia Group B and the European Organization for Research and Treatment of Cancer are the most useful of those currently available. These systems rate performance status, age, histology, and hematologic parameters as the best prognostic factors for mesothelioma. Most patients with mesothelioma are not candidates for surgical or radiotherapy treatment, and cytotoxic agents are the only options. Historically, no classes or combinations of agents consistently yielded response rates over 20%. Recently, pemetrexed has been evaluated in phase I, II, and III clinical trials with promising results. The phase II trial showed an overall response rate of 14.1% with a 1-year survival rate of 47.8%. A phase III trial of cisplatin versus cisplatin and pemetrexed closed in February 2002 and a final analysis was presented in May 2002. Novel targeted agents are also being tested in clinical trials among mesothelioma patients and include drugs inhibiting the vascular endothelial growth factor and its receptor, the epidermal growth factor receptor, and platelet-derived growth factor receptor. PMID- 12571810 TI - Pemetrexed in the treatment of non-small cell lung cancer. AB - A large meta-analysis showed that chemotherapy results in significant prolongation of survival for patients with non-small cell lung cancer (NSCLC). Pemetrexed is a new multitargeted antifolate chemotherapeutic agent that has shown activity in NSCLC and appears to be similar to the third-generation antitumor agents currently in use today. In two phase I trials of pemetrexed in previously untreated patients with NSCLC, overall response rates of 16% and 23% were seen. The treatment was very well tolerated, particularly when dexamethasone premedication was used to prevent rash. In two phase II trials of pemetrexed and cisplatin, overall response rates rose to 39% and 45% with 1-year survival rates of 49% and 50%. Furthermore, in one of the trials, the response rate for stage IV patients was 45%. In the second-line treatment of NSCLC, 9% of patients responded to pemetrexed. The response rate in patients who had not previously received a cisplatin regimen was 14%, and 5% for patients previously treated with a platinum compound. A large randomized trial that compares treatment with pemetrexed to docetaxel in the second-line setting for NSCLC has recently completed accrual. In conclusion, pemetrexed is an active agent in the treatment of advanced NSCLC and represents a new option for the treatment of patients with this malignancy. PMID- 12571811 TI - Pemetrexed in pancreatic cancer. AB - New drugs are clearly needed for the treatment of advanced pancreatic cancer, a disease refractory to most chemotherapy. Pemetrexed, a novel antifolate, inhibits thymidylate synthase, dihydrofolate reductase, glycinamide ribonucleotide formyltransferase, and aminoimidazole carboxamide ribonucleotide formyltransferase. Pemetrexed is active against pancreatic cancer cell lines in vitro. Two partial responses in pancreatic cancer patients were observed in a phase I trial of pemetrexed. This led to a phase II trial of pemetrexed in patients with advanced pancreatic cancer. The objective response rate was 6%, 1 year survival rate was 28%, and the toxicities of therapy were mild. Pemetrexed is synergistic with gemcitabine in vitro. In a phase I trial, the pemetrexed/gemcitabine combination was broadly active and well tolerated. A phase II trial of this combination in 42 patients with advanced pancreatic cancer showed promising activity. A 520-patient international randomized phase III trial that compares the pemetrexed/gemcitabine combination with single-agent gemcitabine is currently accruing patients. PMID- 12571812 TI - The role of pemetrexed in the treatment of colorectal cancer. AB - Advanced colorectal cancer is a leading cause of cancer-related morbidity and mortality. Chemotherapeutic options, however, have recently expanded with concomitant improvements in survival. Through the 1980s and early 1990s research focused mainly on the major fluoropyrimidine, 5-fluorouracil, a thymidylate synthase inhibitor, and methods to enhance its activity through scheduling changes or by biochemical modulation. Pemetrexed is a novel antifolate that inhibits several folate-dependent enzymes in addition to thymidylate synthase. This agent has theoretical and preclinical advantages over fluoropyrimidines and specifically acting antifolates. Phase II studies have shown a broad spectrum of activity in solid tumors, including colorectal cancer. Further studies at higher doses with the use of vitamin supplementation may be desirable. Combinations of pemetrexed with irinotecan and oxaliplatin have also proven feasible. Pemetrexed is a promising new drug for the treatment of colorectal cancer. PMID- 12571813 TI - Pemetrexed: an active new agent for breast cancer. AB - Pemetrexed is a novel antifolate that inhibits three enzymes in the de novo purine and pyrimidine pathways including thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyltransferase. It becomes highly polyglutamated once inside cancer cells, resulting in prolonged intracellular retention and 60-fold greater inhibition of thymidylate synthase than the monoglutamated form. Several phase II and III studies have shown that pemetrexed has significant antitumor activity against a variety of tumor types including mesothelioma, non-small cell lung cancer, and colon, pancreatic, and breast cancers. Pemetrexed appears to provide non-cross-resistant cytotoxic activity against breast cancers that have been treated with anthracyclines, taxanes, and capecitabine. Preclinical studies have suggested that pemetrexed enhances the cytotoxicity of several other important chemotherapeutic agents active against breast cancer including doxorubicin, paclitaxel, cisplatin, and gemcitabine. In vitro studies have shown that pemetrexed treatment synchronizes cancer cells at the G(2)/S interphase, leading to synchronous entry into S phase. Ongoing phase II studies of pemetrexed combinations in breast cancer hope to exploit the cell cycle modulatory effects of this drug as well as its excellent tolerability. PMID- 12571814 TI - Pemetrexed in gastric cancer: clinical experience and future perspectives. AB - The development of more effective and convenient chemotherapy regimens for the treatment of gastric cancer that incorporate novel agents remains an exciting area of research. A phase II study was conducted to assess the response rate and toxicity profile of pemetrexed, a novel multitargeted antifolate, in previously untreated patients with measurable, advanced, or metastatic adenocarcinoma of the stomach or gastroesophageal junction. In this study, pemetrexed-induced toxicity at the starting dose of 500 mg/m(2) intravenously once every 21 days was considerable with each of the first six patients who experienced at least one episode of grade 3/4 toxicity. Two patients discontinued from study, and two patients died. All deaths were caused by drug-related toxicity. No responses were seen in this briefly treated group. These observations led to an amended study protocol designed to improve tolerability of pemetrexed with folic acid supplementation. Supplementation with folic acid 5 mg was given orally once daily for 2 days before pemetrexed on the day of treatment, and for 2 days following treatment. Tumor evaluation was performed after every two cycles of therapy. The trial was recently closed to accrual and preliminary clinical results are reported here. Thirty-two patients were enrolled and 30 patients were evaluable for efficacy. A total of 129 courses of pemetrexed were administered, and the median number of courses received per patient was four (range, one to eight courses). Two complete and five partial responses were observed, with four patients experiencing stable disease. In an intent-to-treat analysis, the overall response rate was 22%, and 23% for the evaluable patients. Median duration of response was 4.4 months (range, 3 to 11 months) and median time to treatment failure was 2.6 months (range, 0.5 to 12 months). Of the 32 patients treated, eight experienced grade 4 neutropenia and one had grade 4 thrombocytopenia. The most common nonhematologic toxicities were diarrhea, fatigue, mucositis, nausea and vomiting, skin rash, and reversible abnormalities in liver function. There was no case of nonhematologic grade 4 toxicity. Although the clinical experience with pemetrexed in advanced gastric cancer remains limited, the promising activity observed in this study indicates that combination studies are warranted. In addition, high-dose intermittent oral folic acid given in this study allowed administration of pemetrexed at the dose and schedule explored with a highly satisfactory safety profile and with no apparent compromise in efficacy. This article discusses how pemetrexed may be investigated in future clinical trials in gastric cancer. PMID- 12571815 TI - Pemetrexed in bladder, head and neck, and cervical cancers. AB - Pemetrexed is a novel multitargeted antifolate analog. The drug has shown encouraging activity in a wide range of solid tumors, including cervix, head and neck, and bladder carcinomas, which are the focus of this review. Toxicity, particularly hematologic, is higher in patients with these tumor types than in other populations exposed to pemetrexed. Supplementation with folic acid and vitamin B(12) appears to effectively reduce the incidence of severe toxicity and may optimize the therapeutic index of pemetrexed in patient subsets with poor nutritional status. The role of this agent in the management of these and other tumor types, as a single agent or in combination, shall be determined by randomized phase III studies. PMID- 12571816 TI - Drug treatment in depressed elderly in the Dutch community. AB - OBJECTIVES: In older people, a diagnosis of depression is frequently missed, and proper treatment is subsequently hampered. We investigated antidepressant and benzodiazepine use in an older community sample, and assessed possible risk factors associated with non-treatment in depressed elderly. METHODS: Data were used from the baseline measurements of the Longitudinal Aging Study Amsterdam (LASA). In a random, age and sex stratified community sample of 3107 older Dutch people (55 to 85 years), respondents were screened on depression with the Center for Epidemiologic Studies Depression Scale (CES-D). In the depressed subsample depressive disorder according to DSM-III was assessed using the Diagnostic Interview Schedule (DIS). The use of antidepressants and anxiolytics (benzodiazepines) in the depressed subsample was measured, and associations with age, sex, cognitive impairment, physical health and anxiety symptoms were investigated. RESULTS: Only 16% of the respondents with a major depressive disorder used antidepressants. More than half of them used non-therapeutic dosages. Lower antidepressant use was associated with cognitive impairment. Benzodiazepine use was more likely than antidepressant use, which was especially evident in females in the major depressive disorder group. CONCLUSIONS: Depressed older people were undertreated, particularly when they were cognitively impaired. A high rate of benzodiazepine use was found, particularly in females. PMID- 12571817 TI - The recognition of dementia in "non-EMI" nursing home residents in South East England. AB - OBJECTIVES: To estimate the agreement between nursing staff's recognition of dementia and results of MMSE assessment in a probability sample of non-specialist nursing home residents in South East England, and to identify correlates of disagreement. METHODS: Prospective survey. The most senior nurse on duty was interviewed about each resident sampled, and optionally about their own training and experience. Residents were interviewed using the MMSE, and assessed using the Cornell Scale for Depression in Dementia, the Barthel ADL index, and the Behave AD scale for behavioural problems. RESULTS: 135 nurses were interviewed about 445 residents-116 reported on of the 291 residents scoring 23 or less on the MMSE-34% of these were acknowledged to have dementia. 46.4% of those with MMSE scores of 15 or less were acknowledged to have dementia. "Missed dementia" was associated with higher MMSE and lower Behave-AD scores, and inversely associated with RMN training and private home ownership for profit. It was not associated with training or duration of staff employment. CONCLUSIONS: Most cognitive impairment in non-specialist nursing homes appeared to be unrecognised. This has implications for the prospects of good dementia care in these homes. PMID- 12571818 TI - A comparison of GMS-A/AGECAT, DSM-III-R for dementia and depression, including subthreshold depression (SD)--results from the Berlin Aging Study (BASE). AB - BACKGROUND: Empirical evaluation of the agreement between different diagnostic approaches is crucial for the understanding of epidemiological results in geriatric psychiatry. OBJECTIVES: In this paper, we analyse differences between widely used diagnostic approaches of dementia and depression and offer evidence that diagnostic thresholds vary substantially on quantitative dimensions, but that conceptual and other differences between approaches must also been taken into account. METHODS: In an epidemiological study of n = 516 persons, aged 70 103 years, we compared psychiatric diagnoses of dementia and depression obtained by GMS-A/HAS-AGECAT, DSM-III-R and clinician's diagnoses of subthreshold depression (SD). RESULTS: For depression, cumulative prevalence of clinician's diagnosis (including SD, GMS-A/HAS-AGECAT and DSM-III-R defined forms) was highest, followed by GMS-A/HAS-AGECAT-diagnosis and DSM-III-R, while for dementia DSM-III-R was followed by GMS-A/HAS-AGECAT. Overall agreement between DSM-III-R and GMS-A/HAS-AGECAT was moderate. Adapting thresholds for AGECAT resulted in slightly better diagnostic efficiency. Diagnostic disagreement was found predominantly for cases with intermediate symptom severity, supporting the hypothesis of differing thresholds between DSM-III-R and GMS-A/HAS-AGECAT, while cases with lower or higher symptom severity were similarly seen as cases or non cases. CONCLUSION: Disagreement is not only caused by conceptual differences, but also different thresholds of diagnostic algorithms. Adaptation of threshold levels should be feasible, depending on the purpose of the analysis. PMID- 12571819 TI - Use of the internet and of the NHS direct telephone helpline for medical information by a cognitive function clinic population. AB - BACKGROUND: Internet websites and medical telephone helplines are relatively new and huge resources of medical information ("cybermedicine" and "telemedicine", respectively) accessible to the general public without prior recourse to a doctor. STUDY OBJECTIVES: To measure use of internet websites and of the NHS Direct telephone helpline as sources of medical information by patients and their families and/or carers attending a cognitive function clinic. DESIGN AND SETTING: Consecutive patients seen by one consultant neurologist over a six-month period in the Cognitive Function Clinic at the Walton Centre for Neurology and Neurosurgery, a regional neuroscience centre in Liverpool, UK. RESULTS: More than 50% of patients and families/carers had internet access; 27% had accessed relevant information, but none volunteered this. 82% expressed interest in, or willingness to access, websites with relevant medical information if these were suggested by the clinic doctor. Although 61% had heard of the NHS Direct telephone helpline, only 10% of all patients had used this service and few calls related to the reason for attendance at the Cognitive Function Clinic. CONCLUSIONS: Internet access and use is common in a cognitive function clinic population. Since information from internet websites may shape health beliefs and expectations of patients and families/carers, appropriately or inappropriately, it may be important for the clinic doctor to inquire about these searches. Since most would use websites suggested by the doctor, a readiness to provide addresses for appropriate sites may prove helpful. PMID- 12571820 TI - Sertraline treatment of elderly patients with depression and cognitive impairment. AB - BACKGROUND: There is little information on the efficacy and side effects of antidepressant treatment in elderly patients with combined depression and cognitive impairment without dementia (DEP-MCI), and it is unclear if cognitive performance improves with antidepressant response in these patients. METHODS: In 39 elderly DEP-MCI patients, changes in depression and cognitive impairment were evaluated with open sertraline treatment up to 200 mg/day for 12 weeks. RESULTS: Of the 26 completers, 17 were responders and nine were non-responders. Diagnostic subtype of depression was unrelated to response. ANCOVA on WAIS-R digit symbol percent change scores revealed a significant effect for responder status (F = 5.59, p < 0.03), and age (F = 0.24, p < 0.64) and education (F = 1.64, p < 0.22) were not significant covariates. From pre-trial to post-trial, responders improved in WAIS-R digit symbol percent change scores (Mean -10% SD 24) while non responders declined (Mean 14% SD 18; t = 2.60, p < 0.02). Other neuropsychological measures were unrelated to response. Percent change in HRSD scores showed significant inverse correlations with percent change in several cognitive measures. CONCLUSIONS: DEP-MCI patients showed moderate clinical response to sertraline treatment. When responders were compared to non responders, cognitive improvement was limited to one measure of attention and executive function. Overall, there was little cognitive improvement with antidepressant treatment. The findings indirectly suggest that lack of improvement in cognition following treatment of depression in DEP-MCI patients may be associated with increased risk of meeting diagnostic criteria for dementia during follow-up. PMID- 12571821 TI - The psychometric properties of the Multi-Dimensional Observation Scale for Elderly Subjects (MOSES) in middle aged and older populations of people with mental retardation. AB - OBJECTIVE: To assess the internal consistency, inter-scale correlations and factor structure of the MOSES with older adults with mental retardation. METHOD: A series of outpatients with mental retardation were assessed with the MOSES. 163 middle aged and older adults with mental retardation living in community settings participated. RESULTS: The subscales and total scale of the Multi-Dimension Observation Scale for Elderly Subjects (MOSES) were highly internally consistent. The pattern of correlations between the five scales was very similar to that reported by Dalton et al. (1999). The results of a factor analysis using varimax rotation indicated a three-factor structure corresponding to adaptive behavior, externalizing and internalizing maladaptive behaviors. CONCLUSIONS: The MOSES appears to be was a psychometrically adequate instrument to screen older adults with mental retardation. PMID- 12571822 TI - Validation of a Spanish translation of the CLOX for use in Hispanic samples: the Hispanic EPESE study. AB - OBJECTIVES: Clock drawing tests (CDT) appear to be less vulnerable to linguistic, cultural, or educational bias than traditional dementia screening instruments. We investigated a Spanish language translation of CLOX: an executive CDT, in a community sample of Hispanic elders. METHODS: In-home CLOX evaluations of 1309 Mexican-American elders were reviewed. RESULTS: Both CLOX1 (an executive CDT) and CLOX2 (a constructional CDT) showed good internal consistency (Chronbach's alpha; both alpha = 0.82). Cultural-demographic variables had little effect on CLOX scores. Although language had a significant effect on CLOX1 failure rates, this was not mediated by age, education, acculturation or income. CONCLUSIONS: These results suggest that the Spanish CLOX can be validly administered to community based Hispanic elder samples regardless of education or acculturation. PMID- 12571823 TI - Prevalence of cognitive impairment amongst Singapore's elderly Chinese: a community-based study using the ECAQ and the IQCODE. AB - OBJECTIVES: This study examined the prevalence of cognitive impairment in the elderly Chinese living in the Queenstown district of Singapore using two locally validated cognitive screening instruments: the Elderly Cognitive Assessment Questionnaire (ECAQ), and the Informant Questionnaire on the Cognitive Decline of the Elderly (IQCODE). METHODS: 234 elderly Chinese subjects were randomly selected from the Queenstown district and administered the ECAQ, while their principal relatives were administered the IQCODE. The screening instruments' respective cut-off scores to identify cognitive impairment indicative of dementia were based on local validation work. RESULTS: The prevalence of cognitive impairment was computed based on the percentage of failed scores on the two screening instruments. We found the prevalence of cognitive impairment to be 7.7% by ECAQ (95% Confidence Intervals (CI): 4.6%-10.1%) and 13.2% by IQCODE (95% CI: 9.4%-17.7%). A similar study in 1990 using ECAQ on elderly Chinese from the Henderson district in Singapore showed a cognitive impairment prevalence of 4%. This ECAQ-based variation between the two studies, likely indicates a true difference in prevalence rates between the two communities. On the other hand, the different ECAQ and IQCODE prevalence rates within our Queenstown study probably reflect spectrum bias, with IQCODE detecting earlier stages of dementia than ECAQ. CONCLUSION: We have found the prevalence of cognitive impairment amongst Singapore's elderly to be higher than previously reported. These findings imply that current resources may be inadequate for the effective care and management of the cognitively-impaired elderly in our aging community. PMID- 12571824 TI - Behavioural disturbances in dementia patients and quality of the marital relationship. AB - OBJECTIVES: To investigate the relationship between behavioural problems in patients with dementia and changes in the marital relationship. METHODS: Fifty three spouse caregivers of patients with dementia participated in the study. Questionnaires and interviews were used to examine caregiver perception of changes in the quality of their relationship. Behavioural disturbances in the patient were measured with the NeuroPsychiatric Inventory (NPI). RESULTS: Caregivers experienced a deterioration of their relationship, yet at the same time most felt closer to their spouse now than in the past. Regression analysis revealed that patient behavioural problems were, independent of patient cognitive status or functional impairment, associated with deterioration in the quality of the relationship between patient and caregiver. Patient apathy rather than depressive mood was associated with this deterioration. Apathy diminished the amount and reciprocity of interactions between partners. CONCLUSIONS: These results show that passive behaviour rather than excessive behaviour has most impact on the deterioration of the marital relationship. Intervention programmes should target relationship problems when problem behaviour, especially apathy, is present in patients with dementia. PMID- 12571825 TI - Memory for new information as a cognitive marker of liability to Alzheimer's disease in a high risk group: a research note. AB - BACKGROUND: The development of Alzheimer's disease (AD) is often insidious and there is evidence that pre-morbid neuropsychological deficits exist. OBJECTIVES: To examine aspects of neuropsychological performance as cognitive markers in a group at high risk of developing AD. METHODS: Memory for novel information and verbal fluency were examined in 33 unaffected biological siblings of patients who fulfilled criteria for probable AD, and 22 controls who reported no family history of dementia. RESULTS: Comparisons between siblings and controls revealed significant differences on overall memory performance. This was also the case when siblings were grouped according to whether or not they possessed the apoE epsilon 4 allele. There were no significant differences between siblings and controls on verbal fluency measures. CONCLUSIONS: Detailed measures of memory performance may be clinically useful in groups at a high risk of developing AD. However, further longitudinal research in such high-risk groups is needed before conclusions can be made with confidence. PMID- 12571826 TI - Reported differences in management strategies by primary care physicians and psychiatrists in older patients who are depressed. AB - OBJECTIVE: To compare reported management suggestions by primary care physicians and psychiatry specialists for case vignette examples of old age depression, and to explore further training needs in geriatric depression for both professional groups. DESIGN: Qualitative study using case vignettes in focus groups. Single group training sessions were arranged for 25 primary care physicians and 11 psychiatrists. The same two clinical vignettes were presented at all training sessions. Written management suggestions by participants, group discussion field notes and transcripts of group session videotapes were analysed. RESULTS: Differences emerged in intended depression management. Primary care physicians tended to assess the symptoms as less serious and the situation as less urgent than psychiatrists. Management suggestions given by the psychiatrists included more recommendations of immediate psychiatric treatment and more precise descriptions of medication. Both groups recognized the somatic issues and were willing to assume responsibility for treatment. CONCLUSIONS: Identifying management differences seems to benefit the professional development of both groups. Exploring and discussing the underlying reasoning leading to management differences may be a productive format for primary care doctors and psychiatric specialists to teach and learn together. It may also promote collaboration in caring for the depressed elderly. PMID- 12571827 TI - The attitudes of carers and old age psychiatrists towards the treatment of potentially fatal events in end-stage dementia. AB - BACKGROUND: Deciding how to treat patients with end-stage dementia developing potentially fatal events has long been contentious. Under expected new legislation the role of carers is likely to increase. Old age psychiatrists frequently have to decide between active or palliative approaches to such patients. Little is known concerning the comparative attitudes of carers and old age psychiatrists. This research examined how their attitudes differed. METHODS: A Likert scale attitudinal questionnaire was designed following a literature review and was initially distributed to a pilot group of medical students to aid in establishing reliability and validity. Subsequently it was sent to all members of the local Alzheimer's Society Branch and all the old age psychiatrists in the health region. Its validity and reliability were investigated using factor analysis and test-retest reliability. Between group comparisons were then made using the students t-test and analysis of variance. RESULTS: 148 carers and 34 clinicians responded. Factor analysis and test-retest analysis of students' responses validated six questions, which grouped into two factors, relating to active treatment and patient-centred ethics. Between group comparisons revealed significant differences between carers' and clinicians' attitudes in these areas. Clinicians favoured active treatment of potentially fatal events in end-stage dementia less than carers who more significantly valued patient-centred issues such as dying with dignity, the patient's best interests and the patient's wishes. CONCLUSIONS: Old age psychiatrists and carers significantly differed in their attitudes towards treatment of potentially fatal events in end-stage dementia and this may impact on decision-making. Considerably more clinician/carer debate and dialogue seems needed. PMID- 12571828 TI - Prevalence and correlates of behavioral and psychiatric symptoms in community dwelling elders with dementia or mild cognitive impairment: the Memory and Medical Care Study. AB - BACKGROUND: Little is known about the prevalence and correlates of behavioral and psychiatric symptoms of dementia in community-dwelling elders with dementia or mild cognitive impairment (MCI). METHODS: 512 people with Mini-Mental State Examination (MMSE) scores < 24 or a decline of at least 4 points over two administrations, and their knowledgeable informants (KIs) were enrolled in the MMCS. The classification of subjects as having dementia or MCI was based on a neuropsychological battery of four tests, not a clinical diagnostic evaluation. The sample for this study included 454 subjects (dementia n = 333; MCI n = 121) and their KIs. Demographic and health-related characteristics of subjects and KIs were obtained during KI interviews. Multivariate logistic regression was used in statistical analysis. RESULTS: Compared to dementia subjects, those classified as MCI had a lower prevalence (47.1% vs 66.1%) of any symptoms (psychosis, depression, or agitation), and of agitation (24.8% vs 45.1%). Symptoms of psychosis and depression also were less prevalent, even though differences did not reach statistical significance. In the dementia group symptoms were associated with a report of a physician's diagnosis of dementia, greater functional impairment, and a KI who was a child/child-in-law. In those with MCI, symptoms were correlated with being white, greater functional impairment, and a younger, less educated, KI. CONCLUSIONS: Psychiatric and behavioral symptoms were common in community-residing elders with cognitive impairment, but their prevalence and correlates differed by study classification as having dementia or MCI. Identifying and treating these symptoms may benefit patients with cognitive impairment and their families. Longitudinal studies on the predictors, changes in prevalence, and effectiveness of treatments for psychopathology of dementia are needed. PMID- 12571829 TI - Symptoms of depression and anxiety predict mortality in elderly veterans enrolled in the UPBEAT program. PMID- 12571830 TI - Medication-induced emergency hospitalizations for psychiatric disorders among older adults in the US. PMID- 12571831 TI - Folic acid and Alzheimer's disease. PMID- 12571832 TI - Paradox revisited: still no direct connection between hours of care and caregiver burden. PMID- 12571833 TI - Effects of the September 11th attacks on symptoms of PTSD on community-dwelling older persons in the Netherlands. PMID- 12571834 TI - Power Doppler sonography: improving disease activity assessment in inflammatory musculoskeletal disease. PMID- 12571835 TI - Improving quality of care in orthopedic surgery. PMID- 12571836 TI - Macrophage migration inhibitory factor: an emerging therapeutic target in rheumatoid arthritis. PMID- 12571837 TI - Anti-double-stranded DNA antibodies, nucleosomes, and systemic lupus erythematosus: a time for new paradigms? PMID- 12571838 TI - Eligibility of patients in routine care for major clinical trials of anti-tumor necrosis factor alpha agents in rheumatoid arthritis. AB - OBJECTIVE: To identify the proportion of patients with rheumatoid arthritis (RA) in 2 cohorts from Nashville, Tennessee, who met basic criteria for inclusion in 2 important recent clinical trials of anti-tumor necrosis factor alpha (anti TNFalpha) agents, the early RA (ERA) trial of etanercept versus methotrexate, and the anti-TNFalpha trial in RA with concomitant therapy (ATTRACT) study of infliximab plus methotrexate versus methotrexate. METHODS: Two cohorts of patients, all of whom had met the American College of Rheumatology criteria for RA at some time, were studied. Cohort E (early) comprised 232 patients who were under the care of 5 private practice rheumatologists, whose duration of RA was fewer than 3 years, and who were reviewed for basic inclusion criteria for the ERA clinical trial. Cohort L (long-term) comprised 152 consecutive patients who had been under care at a weekly academic rheumatology clinic for a mean of 4.5 years, and were reviewed for basic inclusion criteria for the ATTRACT study. RESULTS: In cohort E, basic inclusion criteria for the ERA trial were met by 11 of 36 patients (31%) who had not taken methotrexate, 8 of 19 patients (42%) who were at their first visit and had not taken methotrexate, and 37 of all 232 patients (16%). In cohort L, 5% of patients met the basic inclusion criteria for the ATTRACT study. CONCLUSION: Most patients who were seen in routine care in these 2 cohorts did not meet the criteria for inclusion in these 2 important recent clinical trials. The conclusion that anti-TNFalpha therapy has greater efficacy than methotrexate may be valid only in a limited number of patients with the most severe RA. Anti-TNFalpha therapy may be desirable in most patients with RA, but this possibility has not been studied formally. Criteria for inclusion in RA clinical trials might be modified for greater generalizability of results. PMID- 12571839 TI - Listeria monocytogenes infection as a complication of treatment with tumor necrosis factor alpha-neutralizing agents. AB - OBJECTIVE: Tumor necrosis factor alpha (TNFalpha) has been implicated in the pathogenesis of certain inflammatory diseases. Two TNFalpha-neutralizing agents are licensed in the US. Infliximab is licensed for the treatment of Crohn's disease (CD) and, when used with methotrexate, for the treatment of rheumatoid arthritis (RA). Etanercept is licensed for the treatment of RA, including juvenile RA, and, more recently, was licensed for the treatment of psoriatic arthritis. Because of the potential for decreased host resistance to infectious agents due to treatment with anti-TNFalpha agents, we sought to evaluate postlicensure cases of opportunistic infection, including Listeria monocytogenes, in patients treated with these products. METHODS: The FDA Adverse Event Reporting System, a passive monitoring system, was reviewed to identify all reports of adverse events (through December 2001) associated with L monocytogenes infection in patients treated with infliximab or etanercept. RESULTS: Fifteen cases of L monocytogenes infection associated with infliximab or etanercept treatment were identified. In 14 of these cases, patients had received infliximab. The median age of all patients was 69.5 years (range 17-80 years); 53% were female. Six deaths were reported. Among patients for whom an indication for use was reported, there were 9 patients (64%) with RA and 5 patients (36%) with CD (information was not reported for 1 patient). All patients for whom information was reported were receiving concurrent immunosuppressant drugs. CONCLUSION: Postlicensure surveillance suggests that L monocytogenes infection may be a serious complication of treatment with TNFalpha-neutralizing agents, particularly infliximab. PMID- 12571840 TI - Diagnostic value of high-resolution B-mode and doppler sonography for imaging of hand and finger joints in rheumatoid arthritis. AB - OBJECTIVE: High-resolution sonography enables a detailed assessment of intraarticular and extraarticular soft tissue abnormalities of joints affected by rheumatoid arthritis (RA). This study was undertaken to evaluate the diagnostic value of B-mode sonography and power Doppler compared with that of clinical examinations and conventional radiography. METHODS: The study group comprised 47 patients (14 men, 33 women) with different grades of RA; 31 patients were rheumatoid factor (RF) positive, and 16 were RF negative. The wrists, first through fifth metacarpophalangeal joints, and second through fifth proximal interphalangeal joints of these patients were scored with ultrasound in B-mode and power Doppler application, using a standardized technique. Involvement and severity of inflammation, as well as vascularization, were scored according to a new 3-point scale. The results were correlated with benchmarks of the clinical and radiologic investigations. Clinical status and conventional radiologic status were determined according to the Disease Activity Score and the Larsen score. RESULTS: After preliminary studies in 15 patients, 39% of 704 joints were found to be abnormal by clinical investigation. Erosions were detected by radiography and sonography in 23% and 43% of joints, respectively. Hypervascularization was observed in 34% of 704 joints by power Doppler application. There was a significant correlation (P < 0.001) between the different methods for the detection of the severity of lesions. Use of a modern, state-of-the-art power Doppler program was necessary for semiquantification, and a standardized investigation technique and scoring system provided sufficient quality measures. CONCLUSION: Sonography detects 20% more abnormalities than does radiography, and sonography has the potential to provide simple grading of disease activity. The rate of detection of abnormalities was slightly higher with clinical examination compared with sonography. PMID- 12571841 TI - Cytokine profile of the rheumatoid nodule suggests that it is a Th1 granuloma. AB - OBJECTIVE: To define the cytokine profile within rheumatoid subcutaneous nodules, and to determine whether the destructive inflammatory process in this lesion displays features of a lymphocyte-driven Th1 or Th2 granuloma. METHODS: Subcutaneous nodules excised from 10 patients with rheumatoid arthritis were examined. Transcripts for interleukin 1beta (IL-1beta) IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-15, IL-18, and for tumor necrosis factor alpha (TNFalpha) and interferon-gamma (IFNgamma) were detected by reverse transcription-polymerase chain reaction of extracted RNA. RESULTS: Nine of 10 nodules contained transcripts for IFNgamma. We observed no evidence for the expression of IL-2, IL 4, or IL-5 among the lymphokine genes analyzed. Transcripts for TNFalpha, IL 1beta, IL-10, IL-15, and IL-18 were present in all 10 nodules. Transcripts for IL 12 were present in all but one nodule. Expression of IL-13 messenger RNA was observed in only 5 nodules. CONCLUSION: The cytokine profile within the rheumatoid nodule (i.e., presence of IFNgamma but not IL-2, and prominent expression of IL-1beta and TNFalpha together with IL-12, IL-18, IL-15, and IL-10) is similar to the profile of cytokines in the synovial lesion of rheumatoid arthritis, which is generally accepted as being attributable to a Th1-mediated inflammatory mechanism. Our results suggest that damage to affected synovial membrane or subcutaneous tissue is caused by the same inflammatory mechanisms, and that the nodule is a Th1 granuloma. PMID- 12571843 TI - Transcription factor early growth response 1 activity up-regulates expression of tissue inhibitor of metalloproteinases 1 in human synovial fibroblasts. AB - OBJECTIVE: To investigate the regulatory potential of early growth response 1 (Egr-1) on tissue inhibitor of metalloproteinases 1 (TIMP-1) expression in synovial fibroblasts. METHODS: Egr-1 and TIMP-1 transcripts were detected by in situ hybridization in synovial tissue. Egr-1-regulated TIMP expression was studied in immortalized fibroblast lines using gel retardation assays, RNase protection analysis, reporter gene studies using the human TIMP-1 promoter, and by enzyme-linked immunosorbent assay. RESULTS: TIMP-1 and Egr-1 were coexpressed in synovial fibroblasts of inflamed joints, and Egr-1 activated the expression of TIMP-1. Egr-1 binding to a recognition sequence in the TIMP-1 promoter was demonstrated in gel retardation and reporter gene assays. Since the same DNA sequence was also recognized by the transcription factor Sp-1, our results suggest that the expression of TIMP-1 in synovial fibroblasts may be differentially regulated by Egr-1 and Sp-1. In addition, fibroblasts expressing Egr-1 at high levels were found to express increased levels of TIMP-2 and TIMP-3 messenger RNA. CONCLUSION: The enhanced expression of Egr-1 may regulate the activity of matrix metalloproteinases in synovial fibroblasts by enhancing the expression of the TIMP-1, -2, and -3 genes. PMID- 12571842 TI - Association of interleukin-18 expression with enhanced levels of both interleukin 1beta and tumor necrosis factor alpha in knee synovial tissue of patients with rheumatoid arthritis. AB - OBJECTIVE: To examine the expression patterns of interkeukin-18 (IL-18) in synovial biopsy tissue of patients with rheumatoid arthritis (RA), and to determine whether expression of this primary cytokine is related to the expression of other cytokines and adhesion molecules and related to the degree of joint inflammation. METHODS: Biopsy specimens of knee synovial tissue either without synovitis (n = 6) or with moderate or severe synovitis (n = 11 and n = 12, respectively) were obtained from 29 patients with active RA. Paraffin embedded, snap-frozen sections were used for immunohistochemical detection of IL 18, tumor necrosis factor alpha (TNFalpha), IL-1beta, IL-12, and IL-17. Furthermore, adhesion molecules, such as intercellular adhesion molecule 1, vascular cell adhesion molecule 1, and E-selectin, and cell markers CD3, CD14, and CD68 were stained. RESULTS: IL-18 staining was detectable in 80% of the RA patients, in both the lining and sublining of the knee synovial tissue. IL-18 expression in the synovial tissue was strongly correlated with the expression of IL-1beta (in the sublining r = 0.72, in the lining r = 0.71; both P < 0.0001) and TNFalpha (in the sublining r = 0.59, P < 0.0007, and in the lining r = 0.68, P < 0.0001). In addition, IL-18 expression in the sublining correlated with macrophage infiltration (r = 0.64, P < 0.0007) and microscopic inflammation scores (r = 0.78, P < 0.0001), and with the acute-phase reaction as measured by the erythrocyte sedimentation rate (r = 0.61, P < 0.0004). Interestingly, RA synovial tissue that coexpressed IL-18 and IL-12 demonstrated enhanced levels of the Th1-associated cytokine IL-17. CONCLUSION: Our results show that expression of IL-18 is associated with that of IL-1beta and TNFalpha and with local inflammation in the synovial tissue of patients with RA. In addition, synovial IL 18 expression correlates with the acute-phase response. These data indicate that IL-18 is a primary proinflammatory cytokine in RA that drives the local production of IL-1beta and TNFalpha. PMID- 12571844 TI - Expression of the dendritic cell-associated C-type lectin DC-SIGN by inflammatory matrix metalloproteinase-producing macrophages in rheumatoid arthritis synovium and interaction with intercellular adhesion molecule 3-positive T cells. AB - OBJECTIVE: To determine whether matrix metalloproteinase (MMP)-producing inflammatory macrophages in the synovium of rheumatoid arthritis (RA) patients express the novel dendritic cell (DC)-specific C-type lectin DC-SIGN and whether this expression is associated with the presence of naive T cells expressing the DC-SIGN ligand, intercellular adhesion molecule 3 (ICAM-3). METHODS: Reverse transcription-polymerase chain reaction was performed to study the expression of DC-SIGN in synovium samples from RA, osteoarthritis (OA), and trauma patients. DC SIGN expression on RA monocytes or on monocytes stimulated with granulocyte macrophage colony-stimulating factor and interleukin-4 was further investigated by flow cytometry. To localize DC-SIGN in the synovium, the DC markers ICAM-3 and MMP-1 were analyzed by immunohistochemistry (single and double labeling) on serial cryostat sections. RESULTS: Seventy percent of the inflammatory cells in the synovium of RA patients showed high expression of DC-SIGN. DC-SIGN was expressed by 80% of CD68-positive macrophages, but not by CD83-positive, DC-LAMP positive, or Fascin-positive cells. Normal numbers of DC-SIGN-positive cells were found in the peripheral blood of RA patients, suggesting that DC-SIGN is up regulated locally in the joint. In RA synovium, ICAM-3-positive resting T cells were found in close proximity to DC-SIGN cells. Unexpectedly, a lower percentage of DC-SIGN-expressing cells was found in OA synovium compared with RA synovium. Furthermore, ICAM-3-expressing T cells, which are known to bind DC-SIGN, were almost absent within the synovium of OA and trauma patients. DC-SIGN-positive macrophages adjacent to these T cells were located in close proximity to the cartilage-degrading proteins extracellular MMP inducer (EMMPRIN) and MMP-1. CONCLUSION: The C-type lectin DC-SIGN is almost absent in the synovium of trauma patients but is highly expressed by most CD68-positive macrophages in the synovium of RA patients. The lack of correlation between DC-SIGN expression and the expression of CD83, DC-LAMP, or Fascin indicates that multiple DC/macrophage subsets are present in RA synovium. Expression of DC-SIGN and its ligand, ICAM-3, is found in substantial amounts only in RA synovium, suggesting that their interaction is implicated in the additional activation of synovial macrophages that leads to the production of EMMPRIN and MMP-1. PMID- 12571845 TI - Safety and efficacy of long-term intraarticular steroid injections in osteoarthritis of the knee: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To evaluate the safety and efficacy of long-term intraarticular (IA) steroid injections for knee pain related to osteoarthritis (OA). METHODS: In a randomized, double-blind trial, 68 patients with OA of the knee received IA injections of triamcinolone acetonide 40 mg (34 patients) or saline (34 patients) into the study knee every 3 months for up to 2 years. The primary outcome variable was radiologic progression of joint space narrowing of the injected knee after 2 years. Measurements of minimum joint space width were performed by an automated computerized method on standardized fluoroscopically guided radiographs taken with the patient standing and with the knee in a semiflexed position. The clinical efficacy measure of primary interest was the pain subscale from the Western Ontario and McMaster Universities OA Index (WOMAC). Efficacy measures of secondary interest were the total score on the WOMAC, physician's global assessment, patient's global assessment, patient's assessment of pain, range of motion (ROM) of the affected knee, and 50-foot walking time. Clinical symptoms were assessed just before each injection. RESULTS: At the 1-year and 2-year followup evaluations, no difference was noted between the two treatment groups with respect to loss of joint space over time. The steroid-injected knees showed a trend toward greater symptom improvement, especially at 1 year, for the WOMAC pain subscale, night pain, and ROM values (P = 0.05) compared with the saline injected knees. Using area under the curve analyses, knee pain and stiffness were significantly improved throughout the 2-year study by repeated injections of triamcinolone acetonide, but not saline (P < 0.05). CONCLUSION: Our findings support the long-term safety of IA steroid injections for patients with symptomatic knee OA. No deleterious effects of the long-term administration of IA steroids on the anatomical structure of the knee were noted. Moreover, long-term treatment of knee OA with repeated steroid injections appears to be clinically effective for the relief of symptoms of the disease. PMID- 12571846 TI - Measurement of radiographic joint space width in the tibiofemoral compartment of the osteoarthritic knee: comparison of standing anteroposterior and Lyon schuss views. AB - OBJECTIVE: To evaluate progression of joint space narrowing in radiographs of osteoarthritic (OA) knees imaged in both the standing anteroposterior (AP) and the Lyon schuss positions, using alternative methods to measure joint space width (JSW). METHODS: Standing AP (extended view) and Lyon schuss (posteroanterior [PA] view, with 20-30 degrees of flexion) radiographic images of 58 OA knees were obtained twice (at baseline and 2 years later). With both methods, fluoroscopy was used to align the anterior and posterior margins of the medial or lateral tibial plateau with the central x-ray beam. Minimum JSW, mean JSW, and joint space area (JSA) of the medial or lateral femorotibial joint space were measured using a new digital image analysis system. The effects of knee flexion versus extension and parallel versus nonparallel tibial plateau alignment were evaluated with respect to the reproducibility of JSW in repeated examinations (intraclass correlation coefficient [ICC]), the mean of within-knee standard deviations of repeated measurements (SD(m)), and the sensitivity to changes in JSW in serial radiographs (standardized response mean [SRM]). RESULTS: The performance of the new software, as assessed by the reproducibility of repeated measurements of minimum JSW on the same image, was excellent in both the standing AP (ICC = 0.98) and Lyon schuss radiographs (2 SD(m) = 0.5 mm, ICC = 0.98). The reproducibility in different radiographs of the same knee was not evaluated. However, over 2 years, the mean (+/- SD) decrease in the minimum JSW of OA knees was 0.17 +/- 0.75 mm in standing AP radiographs (P not significant) and 0.24 +/- 0.50 mm in Lyon schuss views (P = 0.007), with SRMs of 0.23 and 0.48, respectively. The quality of alignment of the tibial plateau was satisfactory (<1 mm between anterior and posterior margins of the medial tibial plateau) in 66% of the pairs of Lyon schuss radiographs and in 57% of the pairs of standing AP radiographs. In the Lyon schuss radiographs, SRM was highly dependent on tibial plateau alignment. Minimum JSW was more sensitive to change than was mean JSW or JSA, in paired Lyon schuss radiographs that exhibited satisfactory alignment. CONCLUSION: Compared with the standing AP radiograph, PA imaging of the knee in 20-30 degrees flexion (the schuss position) increases the reproducibility of radiographic JSW measurements in OA knees and the sensitivity to change in JSW in serial radiographs. Sensitivity to change in minimum JSW is notably increased by aligning the medial tibial plateau with the central x-ray beam in the Lyon schuss radiograph. PMID- 12571847 TI - Detection of radiographic joint space narrowing in subjects with knee osteoarthritis: longitudinal comparison of the metatarsophalangeal and semiflexed anteroposterior views. AB - OBJECTIVE: Although recent protocols for standardized knee radiography afford highly reproducible radioanatomic alignment of the joint and measurement of joint space width (JSW) in repeat radiographs acquired on the same day, the sensitivity of these techniques to joint space narrowing (JSN) over time in subjects with knee osteoarthritis (OA) is unknown. The present study was undertaken to compare the metatarsophalangeal (MTP) view and the semiflexed anteroposterior (AP) view with respect to sensitivity to JSN in knee OA. METHODS: In 49 subjects with definite knee OA, 2 MTP radiographs and 1 semiflexed AP radiograph were obtained at baseline. Each examination was repeated 14 months later. In MTP views, minimum JSW and the distance between the anterior and posterior margins of the medial tibial plateau (intermargin distance [IMD], an indicator of parallel alignment of the tibial plateau and the x-ray beam) were measured with a pair of calipers and a magnifying lens fitted with a graticule. JSW in semiflexed AP views was measured by digital image analysis. RESULTS: The mean of within-knee standard deviations of JSW in the baseline MTP examinations (n = 52 OA knees) was 0.24 mm (coefficient of variation 5.8%). Although IMDs in the 2 baseline MTP views were very highly correlated (+0.88), IMDs in the serial examinations were only moderately correlated (+0.45). Serial MTP views showed a small increase in mean JSW over 14 months that was not significantly greater than zero (mean +/- SD +0.09 +/- 0.66 mm; P not significant). In contrast, concurrent semiflexed AP examinations showed a marginally significant decrease in mean JSW (-0.09 +/- 0.31 mm; P = 0.10). CONCLUSION: These results demonstrate that evidence of the short term reproducibility of a radiographic protocol is an insufficient basis on which to predict the quality of its longitudinal performance. PMID- 12571848 TI - The inheritance of hand osteoarthritis in Iceland. AB - OBJECTIVE: To assess the contribution of genetics to hand osteoarthritis (HOA) and its subsets in the Icelandic population. METHODS: A list of 2,919 HOA patients, constituting 1% of the Icelandic population, was compiled through nationwide sources. This patient list was cross-referenced with a comprehensive Icelandic genealogy database, enabling the use of algorithms to assess familiality of HOA. Two methods were used: the average pairwise kinship coefficient (KC) of the patients, and the relative risk (RR) of HOA in relatives of patients. In each case, the results were compared with 1,000 control sets of similar composition with regard to number, age, and sex, generated from the genealogy database. RESULTS: The KC for patients was significantly higher than for the control sets and was proportional to the degree of both interphalangeal (IP) and thumb base (first carpometacarpal [CMC] joint) involvement. The RR of HOA in sisters of women in the study was 2.0 (P < 0.001), while the RR in spouses was not significantly different from that in controls. The RR increased with the severity of the disease. Thus, sisters of women with severe IP HOA had an RR of 5.0 and sisters of those with severe first CMC involvement had an RR of 6.9. The increased risk also extended beyond the nuclear family, with significantly increased risk in cousins. CONCLUSION: Patients seeking medical services for HOA are more related to each other than matched controls, supporting the role of a genetic component in the disease. The genetic influence in both IP and first CMC HOA appears to be similar and increases with increasing severity of the disease. PMID- 12571849 TI - Immature articular cartilage is more susceptible to blood-induced damage than mature articular cartilage: an in vivo animal study. AB - OBJECTIVE: Cartilage of young but skeletally mature dogs is more susceptible to blood-induced damage than that of old dogs. The aim of the present study was to investigate whether cartilage of skeletally immature individuals is even more adversely affected by exposure to blood than that of mature individuals, as suggested by clinical practice experience with humans. METHODS: Right knees of 3 groups of 6 beagle dogs (skeletally immature, young mature, and old animals) were injected with autologous blood on days 0 and 2. On day 4, cartilage matrix proteoglycan turnover (content, synthesis, and release), synovial inflammation, and cartilage-destructive properties of the synovial tissue were determined and compared with those of the left uninjected control knees. RESULTS: Subsequent to intraarticular bleeding, cartilage proteoglycan content decreased in an age dependent manner, with the largest decrease occurring in cartilage of immature animals. Proteoglycan synthesis per cell also decreased in an age-dependent manner, with the largest decrease occurring in the immature animals. Cartilage proteoglycan release increased in all 3 groups, but the decrease was not age dependent. Interestingly, immature animals showed a large increase in cartilage DNA content upon exposure to blood, whereas mature animals did not. Histologic analysis showed a mild synovitis in animals of all ages, but catabolic inflammatory activity was found only in immature animals. CONCLUSION: Joints of skeletally immature dogs appeared to be more susceptible than joints of mature dogs to the adverse effects of a joint hemorrhage. These data suggest that for humans, specifically young children are at risk for joint damage after a joint hemorrhage. PMID- 12571850 TI - Synergistic induction of nitric oxide synthase type II: in vitro effect of leptin and interferon-gamma in human chondrocytes and ATDC5 chondrogenic cells. AB - OBJECTIVE: To study, in vitro, the effect of leptin (OB), alone or in combination with interferon-gamma (IFNgamma), on inducible nitric oxide synthase (iNOS) and NO production in human primary chondrocytes and in mouse embryonic chondrogenic ATDC5 cells. METHODS: Leptin receptor expression and iNOS messenger RNA expression were evaluated by reverse transcriptase-polymerase chain reaction. Then, iNOS activity was indirectly studied by measuring nitrite accumulation, using the Griess colorimetric reaction, in culture medium of human primary chondrocytes and ATDC5 cells. RESULTS: ATDC5 mouse embryonic cells expressed functional OB receptor. Alone, neither OB nor IFNgamma produced nitrite accumulation in culture medium. However, costimulation with OB and IFNgamma resulted in dose-dependent up-regulation of the expression of iNOS and NO production in human primary chondrocytes and ATDC5 cells. Production of NO was blunted by the iNOS-specific inhibitors L-N(G)-nitroarginine methyl ester and aminoguanidine. In addition, the janus-activated kinase 2 (JAK2)-specific inhibitor Tyrphostin AG 490 completely blocked OB + IFNgamma-driven up-regulation of iNOS and NO production. CONCLUSION: Our data show for the first time a putative proinflammatory role of OB via iNOS induction and NO production. This occurs via activation of JAK2. PMID- 12571851 TI - Synthesis of insulin-like growth factor binding protein 3 in vitro in human articular cartilage cultures. AB - OBJECTIVE: To quantify the rate of synthesis of insulin-like growth factor binding protein 3 (IGFBP-3) and insulin-like growth factor 1 (IGF-1) by in vitro cultures of normal and osteoarthritic (OA) human articular cartilage. METHODS: Levels of IGF-1 and IGFBP-3 in media from in vitro cultures of human cartilage were determined by radioimmunoassay (RIA). IGFBPs were characterized by immunoblots and ligand blots. Ultrafiltration and RIA analysis of synovial fluid (SF) samples and washings of cartilage samples ex vivo were used to calculate partition coefficients and to estimate the amount of IGF-1 and IGFBP-3 in cartilage in vivo. RESULTS: OA cartilage synthesized 150 ng of IGFBP-3 per gm of cartilage per day, compared with 50 ng synthesized by normal cartilage. The surface zone of normal cartilage produced more IGFBP-3 than did the deep zone. Immunoblots and ligand blots confirmed the presence of IGFBP-3. IGFBP-3 synthesis was stimulated by exogenous IGF-1. No freshly synthesized IGF-1 was detected. The quantities of IGF-1 and IGFBP-3 present ex vivo were 11.3 and 78.7 ng/gm of cartilage in normal cartilage and 21.6 and 225.4 ng/gm in OA cartilage. CONCLUSION: The results show that while IGFBP-3 is synthesized in explant cultures, IGF-1 is not. The rate of IGFBP-3 synthesis is 3 times higher in OA than in normal cartilage. Both IGFBP-3 and IGF-1 penetrate into cartilage from SF in vivo. We estimate that the quantities of IGFBP-3 produced in culture by human cartilage are small compared with the amount supplied in the form of "small complexes" from the circulation. The high value of the partition coefficient of IGFBP-3 implies binding to the matrix. PMID- 12571852 TI - Cartilage-like gene expression in differentiated human stem cell spheroids: a comparison of bone marrow-derived and adipose tissue-derived stromal cells. AB - OBJECTIVE: To compare the chondrogenic potential of human bone marrow-derived mesenchymal stem cells (BMSC) and adipose tissue-derived stromal cells (ATSC), because the availability of an unlimited cell source replacing human chondrocytes could be strongly beneficial for cell therapy, tissue engineering, in vitro drug screening, and development of new therapeutic options to enhance the regenerative capacity of human cartilage. METHODS: Quantitative gene expression of common cartilage and cell interaction molecules was analyzed using complementary DNA array technology and reverse transcription-polymerase chain reaction during optimization of cell differentiation, in order to achieve a molecular phenotype similar to that of chondrocytes in cartilage. RESULTS: The multilineage potential of BMSC and ATSC was similar according to cell morphology and histology, but minor differences in marker gene expression occurred in diverse differentiation pathways. Although chondrogenic differentiation of BMSC and ATSC was indistinguishable in monolayer and remained partial, only BMSC responded (with improved chondrogenesis) to a shift to high-density 3-dimensional cell culture, and reached a gene expression profile highly homologous to that of osteoarthritic (OA) cartilage. CONCLUSION: Hypertrophy of chondrocytes and high matrix remodeling activity in differentiated BMSC spheroids and in OA cartilage may be the basis for the strong similarities in gene expression profiles between these samples. Differentiated stem cell spheroids represent an attractive tool for use in drug development and identification of drug targets in OA cartilage-like tissue outside the human body. However, optimization of differentiation protocols to achieve the phenotype of healthy chondrocytes is desired for cell therapy and tissue engineering approaches. PMID- 12571853 TI - Articular cartilage repair by gene therapy using growth factor-producing mesenchymal cells. AB - OBJECTIVE: To investigate the repair of partial-thickness lesions in rat articular cartilage by combining cell transplantation with transfer of growth factor complementary DNA (cDNA). METHODS: Mesenchymal cells isolated from rib perichondrium were infected ex vivo with adenoviral vectors carrying bone morphogenetic protein 2 (BMP-2) or insulin-like growth factor 1 (IGF-1) cDNA. The cells were suspended in fibrin glue and applied to mechanically induced partial thickness cartilage lesions in the patellar groove of the rat femur. The filling of the defects was quantified and the quality and integration of the newly formed tissue were assessed by histochemical and immunohistochemical methods. Uninfected cells or cells infected with a LacZ reporter gene vector served as controls. RESULTS: Transplanted cells were able to attach to the wounded articular cartilage and were not displaced from the lesions by joint movement. Cells infected with both adenoviral vectors AdBMP-2 and AdIGF-1 produced repair cartilage of hyaline morphology containing a type II collagen-positive but type I collagen-negative proteoglycan-rich matrix that restored the articular surface in most lesions. Uninfected cells either failed to fill up the defects or formed fibrous tissue mainly composed of type I collagen. Excessive cells were partially dislocated to the joint margins, leading to osteophyte formation there if AdBMP-2 infected cells were used. These adverse effects, however, were not seen with AdIGF-1-infected cells. CONCLUSION: Stimulation of perichondrium-derived mesenchymal cells by transfer of growth factor cDNA in a partial-thickness defect model allows for satisfactory cartilage restoration by a repair tissue comparable with hyaline articular cartilage. PMID- 12571854 TI - LJP 394 for the prevention of renal flare in patients with systemic lupus erythematosus: results from a randomized, double-blind, placebo-controlled study. AB - OBJECTIVE: To determine whether LJP 394 delays or prevents renal flare in patients with systemic lupus erythematosus (SLE) and a history of renal disease. METHODS: In a 76-week, double-blind, placebo-controlled study, 230 SLE patients were randomized to receive 16 weekly doses of 100 mg of LJP 394 or placebo, followed by alternating 8-week drug holidays and 12 weekly doses of 50 mg of LJP 394 or placebo. An assay measuring the affinity of the serum IgG fraction for the DNA epitope of LJP 394 identified a high-affinity population of patients (189 of 213 patients; 89% taking LJP 394 and 90% taking placebo). Analyses were performed on both the intent-to-treat population and the high-affinity population. RESULTS: Anti-double-stranded DNA antibodies decreased and C3 levels tended to increase during treatment with LJP 394. In the intent-to-treat population, the time to renal flare was not significantly different between treatment groups, but patients taking LJP 394 had a longer time to institution of high-dose corticosteroids and/or cyclophosphamide (HDCC) and required 41% fewer treatments with HDCC. In the high-affinity population, the LJP 394 group experienced a longer time to renal flare, 67% fewer renal flares, longer time to institution of HDCC, and 62% fewer HDCC treatments compared with the placebo group. In patients with serum creatinine levels >/=1.5 mg/dl at study entry, those taking LJP 394 had 50% fewer renal flares; no renal flares were observed in the high-affinity group taking LJP 394. Serious adverse events were observed in 25 of the 114 LJP 394-treated patients (21.9%) and 34 of the 116 placebo-treated patients (29.3%). CONCLUSION: Treatment with LJP 394 in patients with high-affinity antibodies to its DNA epitope prolonged the time to renal flare, decreased the number of renal flares, and required fewer HDCC treatments compared with placebo. The study drug appeared to be well tolerated. PMID- 12571855 TI - The relationship of FcgammaRIIIa genotype to degree of B cell depletion by rituximab in the treatment of systemic lupus erythematosus. AB - OBJECTIVE: Despite wide use of the anti-CD20 monoclonal antibody rituximab in the treatment of B cell lymphomas, the mechanism by which it causes B cell depletion remains a subject of controversy. As part of an ongoing phase I/II trial of rituximab in the treatment of systemic lupus erythematosus (SLE), we sought to determine whether the effectiveness of B cell depletion was influenced by polymorphisms of Fc receptors (FcR) on effector cells. METHODS: During rituximab treatment of 12 SLE patients, B cell depletion was monitored as a function of the serum rituximab level and FcgammaRIIa and FcgammaRIIIa genotypes at baseline and at 1 month and 2 months after treatment. FcR genotypes were determined by polymerase chain reaction. Serum levels of rituximab were measured by enzyme linked immunosorbent assay (ELISA). B lymphocyte percentages were assessed by flow cytometry. RESULTS: B cell depletion was highly variable in this patient cohort, with B cell percentages at the 1-2-month posttreatment nadir ranging from undetectable (<0.1 cell/microl) to 16% ( approximately 30 cells/microl) of the total peripheral blood lymphocytes. At 2 months posttreatment, B cell percentages were highly correlated with both the serum rituximab level and the FcgammaRIIIa genotype (R(2) = 0.75, P = 0.002). The FcgammaRIIIa genotype was a significant independent predictor of the efficacy of B cell depletion (P = 0.019). CONCLUSION: These results highlight the potential variability of B cell depletion by rituximab in the treatment of autoimmune disease and indicate that Fc receptors are an important determinant of that variability. The findings further suggest the importance of antibody-dependent cell-mediated cytotoxicity and/or apoptosis induction via FcgammaRIIIa-expressing effector cells in the mechanism of B cell depletion by this widely used monoclonal antibody. PMID- 12571856 TI - Identification of IgG subclasses and C-reactive protein in lupus nephritis: the relationship between the composition of immune deposits and FCgamma receptor type IIA alleles. AB - OBJECTIVE: To characterize the subclass composition of IgG deposited in lupus glomeruli, to examine its relationship to allelic polymorphisms of IgG receptors (Fcgamma receptors [FcgammaR]), and to determine whether C-reactive protein (CRP), a ligand for FcgammaRIIa, is present in these immune deposits. METHODS: Renal biopsy samples from 80 patients with lupus nephritis were examined by light microscopy and indirect immunofluorescence with IgG-subclass-specific monoclonal antibodies. FcgammaRIIA genotypes were determined using allele-specific polymerase chain reaction. Immunostaining for CRP was performed on lupus and nonlupus glomerulonephritis specimens. RESULTS: IgG2 and IgG3 were the predominant subclasses in immune deposits in all World Health Organization classes of nephritis. The frequency of genotypes containing the low-binding IgG2 allele, FcgammaRIIa-R131, was significantly greater than expected in patients with class III or class IV nephritis and in patients with intense IgG2 deposition. CRP, a ligand with particular affinity for FcgammaRIIa-R131, was consistently present in the renal immune deposits of lupus nephritis specimens. CONCLUSION: FcgammaRIIA genes are associated with proliferative renal disease and may contribute to disease pathogenesis. FcgammaRIIa-R131, the variant with low affinity for IgG2, has high affinity for CRP. Thus, FcgammaRIIa-R131 may contribute to impaired removal of circulating immune complexes, as well as efficiently triggering phagocyte activation and the release of inflammatory mediators within glomeruli. PMID- 12571857 TI - Valine/valine genotype at position 247 of the beta2-glycoprotein I gene in Mexican patients with primary antiphospholipid syndrome: association with anti beta2-glycoprotein I antibodies. AB - OBJECTIVE: To determine the polymorphism at position 247 of the beta(2) glycoprotein I (beta(2)GPI) gene in Mexican patients with antiphospholipid syndrome (APS) and to compare these data in patients with or without antibodies to beta(2)GPI and with the clinical manifestations of APS. METHODS: We studied 39 patients with primary APS and compared them with 106 clinically healthy subjects. Polymorphism was determined by polymerase chain reaction-restriction fragment length polymorphism. The presence of "true" anticardiolipin (aCL) antibodies, beta(2)GPI-dependent aCL antibodies (IgG and IgM), and phospholipid-free anti beta(2)GPI antibodies (IgG isotype) were detected by enzyme-linked immunosorbent assay (ELISA) utilizing nonirradiated ELISA plates. Clinical manifestations associated with antiphospholipid antibodies were also evaluated. RESULTS: We found no significant differences in the genotype expression between the control group and the primary APS patients (13% with VV, 52% with VL, and 35% with LL versus 23% with VV, 51% with VL, and 26% with LL, respectively). In contrast, anti-beta(2)GPI-positive patients had significantly higher frequencies of the VV genotype and V allele expression than the control subjects and the anti beta(2)GPI-negative patients. These genotype and allele frequencies were also significantly higher in patients with arterial thrombosis than in patients without it. Anti-beta(2)GPI-negative patients without arterial thrombosis did not express the VV genotype. We found no differences in the Val/Leu(247) polymorphism of the beta(2)GPI gene in primary APS patients with or without "true" aCL antibodies or in primary APS patients with or without beta(2)GPI-dependent aCL antibodies. CONCLUSION: Our results suggest that the VV genotype at position 247 of the beta(2)GPI gene may play a role in the generation of anti-beta(2)GPI antibodies and perhaps in the expression of arterial thrombosis in primary APS. PMID- 12571858 TI - Identification and characterization of SmD183-119-reactive T cells that provide T cell help for pathogenic anti-double-stranded DNA antibodies. AB - OBJECTIVE: The C-terminal peptide of amino acids 83-119 of the SmD1 protein is a target of the autoimmune response in human and murine lupus. This study was undertaken to test the hypothesis that SmD1(83-119)-reactive T cells play a crucial role in the generation of pathogenic anti-double-stranded DNA (anti dsDNA) antibodies. METHODS: Splenic or lymph node T cells derived from unmanipulated as well as SmD1(83-119)-immunized NZB/NZW mice were analyzed in vitro by enzyme-linked immunospot (ELISpot) assay to determine T cell help for anti-dsDNA generation induced by the SmD1(83-119) peptide. Cytokines expressed by these T cells were measured by ELISpot assay, enzyme-linked immunosorbent assay, and flow cytometry. SmD1(83-119)- and ovalbumin-specific T cell lines were generated and characterized. RESULTS: The SmD1(83-119) peptide, but not the control peptides, significantly increased the in vitro generation of anti-dsDNA antibodies in cultures from unmanipulated NZB/NZW mice. Interferon-gamma (IFNgamma), interleukin-2 (IL-2), IL-4, transforming growth factor beta, and IL 10 production increased in response to the peptide in young mice; only IFNgamma and IL-2 were increased in older, diseased mice. Activation of SmD1(83-119) reactive T cells by immunization of NZB/NZW mice resulted in elevated anti-dsDNA synthesis and, later, increased antibodies to SmD1(83-119). Most cells in SmD1(83 119)-specific CD4+ T cell lines helping both antibodies had increased intracellular expression of IFNgamma, and most expressed both IFNgamma and IL-4. CONCLUSION: The SmD1(83-119) peptide plays an important role in generating T cell help for autoantibodies, including anti-dsDNA, and activates different subsets of T cells as defined by distinct cytokine expression. This peptide is an interesting target structure for the modulation of autoreactive T cells, and its characterization may contribute to our understanding of the role of autoantigen reactive T cells in the pathogenesis of SLE. PMID- 12571859 TI - Fc receptor-independent development of autoimmune glomerulonephritis in lupus prone MRL/lpr mice. AB - OBJECTIVE: To determine the role of Fc receptors (FcR), which play crucial roles in antibody and immune complex-mediated inflammation and autoimmunity, including glomerulonephritis (GN), in the development of autoimmune GN and vasculitis in MRL/lpr mice, one of the most widely used lupus-prone mouse models. METHODS: FcRgamma(-/-) MRL/lpr mice were generated by backcrossing for 8 generations. The development of GN and vasculitis of various sized vessels was analyzed histopathologically in the kidney, lung, and skin. Autoantibody and immune complex levels were determined biochemically at 16-24 weeks of age and compared with the findings in FcRgamma(+) MRL/lpr mice. The lifespan of the mice was also recorded. RESULTS: Diffuse proliferative GN, with deposition of IgG and C3, developed in both FcRgamma(-/-) and FcRgamma(+) MRL/lpr mice. There was no difference in the survival rate and degree of proteinuria between FcRgamma(+) and FcRgamma(-/-) MRL/lpr mice. Regardless of the level of FcR expression, there were no significant differences in the levels of serum IgG, anti-DNA antibody, or circulating immune complexes between the two types of mice. Necrotizing vasculitis in medium-sized arteries of the kidneys and lungs as well as small vessel vasculitis in the skin was observed in both in FcRgamma(+) and FcRgamma(-/ ) MRL/lpr mice. In contrast, the Arthus reaction was induced in FcRgamma(+) MRL/lpr mice, but not in FcRgamma(-/-) MRL/lpr mice. CONCLUSION: Unlike (NZB x NZW)F(1), the other strain of lupus-prone mice that develops GN in an FcR dependent manner, the development of autoimmune GN and vasculitis in MRL/lpr mice was FcR-independent, implying heterogeneity of the contribution of FcR to the development of autoimmune disease. PMID- 12571860 TI - Effects of anti-CD154 treatment on B cells in murine systemic lupus erythematosus. AB - OBJECTIVE: To determine the immunologic effects of anti-CD154 (CD40L) therapy in the (NZB x NZW)F(1) mouse model of systemic lupus erythematosus. METHODS: Twenty week-old and 26-week-old (NZB x NZW)F(1) mice were treated with continuous anti CD154 therapy. Mice were followed up clinically, and their spleens were studied at intervals for B and T cell numbers and subsets and frequency of anti-double stranded DNA (anti-dsDNA)-producing B cells. T cell-dependent immunity was assessed by studying the humoral response to the hapten oxazolone. RESULTS: IgG anti-dsDNA antibodies decreased during therapy and disease onset was delayed, but immune tolerance did not occur. During treatment, there was marked depletion of CD19+ cells in the spleen; however, autoreactive IgM-producing B cells could still be detected by enzyme-linked immunospot assay. In contrast, few IgG- and IgG anti-dsDNA-secreting B cells were detected. Eight weeks after treatment cessation, the frequency of B cells producing IgG anti-dsDNA antibodies was still decreased in 50% of the mice, and activation and transition of T cells from the naive to the memory compartment were blocked. Anti-CD154 treatment blocked both class switching and somatic mutation and induced a variable period of relative unresponsiveness of IgG anti-dsDNA-producing B cells, as shown by decreased expression of the CD69 marker and failure to generate spontaneous IgG anti-dsDNA producing hybridomas. Treated mice mounted an attenuated IgM response to the hapten oxazolone and produced no IgG antioxazolone antibodies. CONCLUSION: Anti CD154 is a B cell-depleting therapy that affects multiple B cell subsets. Activation of both B and T cells is prevented during therapy. After treatment cessation, autoreactive B cells progress through a series of activation steps before they become fully competent antibody-producing cells. The general immunosuppression induced during treatment will need to be taken into account when using B cell-depleting regimens in humans. PMID- 12571861 TI - Radiologic features in juvenile idiopathic arthritis: a first step in the development of a standardized assessment method. AB - OBJECTIVE: To describe radiologic features of patients with juvenile idiopathic arthritis (JIA) in a standardized manner, to test the reliability and feasibility of this description, and to correlate these features with clinical signs as a first step in the development of a standardized assessment method. METHODS: The placebo-controlled study of sulfasalazine in patients with oligoarticular, extended oligoarticular, and polyarticular JIA performed by the Dutch Juvenile Idiopathic Arthritis Study Group yielded the data for this study. All trial entry radiographs (clinically involved joints and contralateral joints) were scored (in consensus by a skeletal radiologist and pediatric rheumatologist) for the presence of swelling, osteopenia, joint space narrowing, growth abnormalities, subchondral bone cysts, erosions, and malalignment. RESULTS: Data on 67 of 69 patients were analyzed. The mean age was 9.1 years (range 2.5-17.6 years), and the median disease duration was 24 months (range 5-176 months). Thirteen percent of the patients were IgM rheumatoid factor (IgM-RF) positive, and 16% were HLA B27 positive. All 68 clinically evaluated joints were included in the maximum of 19 radiographed joints (or joint groups) per patient. The mean number of radiographed joints per patient was 7 (range 2-15); knees, hands, ankles, and feet were most frequently affected. Fifty-eight patients (87%) had radiologic abnormalities in at least one joint (soft-tissue swelling in 63% of patients, growth disturbances in 48%, joint space narrowing in 28%, and erosions in 15%). In total, half of the radiographs of the clinically involved joints showed radiologic abnormalities, including two-thirds of the radiographs of the clinically affected hands and knees. Univariate analysis revealed a good correlation between the overall articular (clinical) severity and the presence of radiologic abnormalities (odds ratio [OR] 1.38, P < 0.0001). Multivariate analysis showed increased ORs for the presence of radiologic abnormalities and IgM-RF positivity (OR 4.6, P = 0.005) or HLA-B27 positivity (OR 3.0, P = 0.004). In general, reproducibility of the radiologic scoring method was good (mean kappa coefficient of 0.74 [range 0.40-0.86]), although there were scoring discrepancies for swelling, osteopenia, and growth disturbances. The scoring took 10-20 minutes per patient. CONCLUSION: Our model of describing and scoring radiologic abnormalities of radiographed joints in JIA was feasible, mostly reproducible, correlated well with the overall articular severity score, and added substantial new information not available on clinical examination. PMID- 12571862 TI - Predictors of isolated pulmonary hypertension in patients with systemic sclerosis and limited cutaneous involvement. AB - OBJECTIVE: To determine whether there are factors, such as the diffusing capacity for carbon monoxide (DLCO) or pulmonary artery pressure (PAP) on echocardiogram, that can predict the development of pulmonary hypertension (PHT) in patients with limited scleroderma. METHODS: Using the large Pittsburgh Scleroderma Databank, 106 patients who had the diagnosis of PHT after January 1, 1982, were matched with 106 controls by scleroderma subtype, age, sex, race, disease duration, and the mean time to the diagnosis of PHT after the initial Pittsburgh visit. Autoantibodies, vascular features, use of calcium channel blockers, extent of pulmonary function, and echocardiogram findings were determined at any time prior to the diagnosis of PHT (or prior to the matched time in controls). RESULTS: Patients with PHT had a mean DLCO of 52% of predicted at an average of 4.5 years prior to the diagnosis of PHT. This was markedly decreased compared with the values in controls, whose mean DLCO was 81% of predicted (P < 0.0001). The estimated mean PAP on echocardiogram was only slightly higher in the PHT patients compared with controls (34 mm Hg versus 29 mm Hg; P not significant). Nineteen PHT patients had 4 serial measurements of the DLCO during the 15 years prior to the diagnosis of PHT. The initial mean DLCO was 80% of predicted, which decreased in a linear manner to a mean of 35% of predicted at the time of diagnosis of PHT, whereas the value in controls remained at approximately 80% of predicted (P < 0.0001). PHT patients had more severe Raynaud's phenomenon and more severe digital tip ulcers, but they used calcium channel blockers significantly less frequently (37% versus 61% of controls; P < 0.01). The predominance of nucleolar autoantibodies and the absence of anti-Scl 70 antibody were associated with PHT. CONCLUSION: A decreasing DLCO is an excellent predictor of the subsequent development of isolated PHT in limited scleroderma. The DLCO may be significantly decreased for many years prior to the diagnosis of PHT. The presence of autoantibodies and the PAP may also be helpful predictors. The long-term use of calcium channel blockers may be protective, but newer agents that are more effective in treating PHT may also be helpful in altering the natural history of this serious complication in limited scleroderma. PMID- 12571863 TI - Assessment of peripheral enthesitis in the spondylarthropathies by ultrasonography combined with power Doppler: a cross-sectional study. AB - OBJECTIVE: To assess the prevalence and severity of peripheral enthesitis among the different subtypes of spondylarthropathy (SpA) by using ultrasonography (US) in B mode with power Doppler. METHODS: One hundred sixty-four consecutive patients with SpA (according to the criteria of the European Spondylarthropathy Study Group) and 64 control patients (34 with mechanical low back pain [MBP] and 30 with rheumatoid arthritis [RA]) underwent US examination of major entheses of their limbs. Particular attention was given to the detection of vascularization at the following sites: cortical bone insertion of entheses, junction between tendon and entheses, body of tendon, and bursa. RESULTS: Abnormal US findings consistent with at least one enthesitis were observed in 161 of 164 SpA patients (98%), affecting 1,131 of 2,952 entheses examined (38%). In contrast, only 132 of 1,152 entheses (11%) were found to be abnormal in 33 of 64 control patients (52%). US enthesitis was most commonly distributed in the distal portion of the lower limbs, irrespective of SpA subtype and of skeletal distribution of clinical symptoms. None of the abnormal entheses in control patients showed vascularization, compared with 916 of 1,131 abnormal entheses in SpA patients (81%), where it was always detected at the cortical bone insertion and sometimes also in the bursa. In SpA patients, the US pattern depended on the clinical presentation, with a higher prevalence of the most severe stages in those with peripheral forms. CONCLUSION: US in B mode combined with power Doppler allowed the detection of peripheral enthesitis in a majority of SpA patients, but not in MBP or RA patients. The presence of entheseal involvement was independent of SpA subtype, but its degree of severity appeared to be greater in peripheral forms. US could be very useful for both the diagnosis and the assessment of SpA activity. PMID- 12571864 TI - Binding of outer surface protein A and human lymphocyte function-associated antigen 1 peptides to HLA-DR molecules associated with antibiotic treatment resistant Lyme arthritis. AB - OBJECTIVE: To assess the binding of outer surface protein A (OspA) and human lymphocyte function-associated antigen 1 (hLFA-1) peptides to 5 major histocompatibility complex (MHC) molecules. METHODS: Peptide binding to the MHC molecules was determined by in vitro binding assays, and binding was correlated with the frequencies of the 5 MHC molecules in patients with treatment-resistant Lyme arthritis. RESULTS: The HLA-DRB1*0401 molecule bound both OspA(163-175) and hLFA-1alpha(L330-342) well. Although the magnitude of the binding was less, the DRB1*0404 molecule also showed binding of both peptides. The DRB1*0101 molecule bound OspA(163-175) well, but hLFA-1alpha(L330-342) only weakly; the DRB1*0801 or *1101 molecule bound both peptides weakly, if at all. The magnitude of OspA(163 175) binding correlated well with the frequencies of the DRB1 alleles in patients with treatment-resistant arthritis, but the binding of hLFA-1alpha(L330-342) showed only an association with the DRB*04 alleles. CONCLUSION: These correlations support the hypothesis that OspA(163-175) is the critical epitope in triggering antibiotic treatment-resistant Lyme arthritis. However, the inability of the DRB*0101 molecule to bind hLFA-1alpha(L330-342) suggests that this peptide may not be a relevant autoantigen, at least in DRB1*0101-positive patients. PMID- 12571866 TI - Mapping and functional characterization of rat chromosome 4 regions that regulate arthritis models and phenotypes in congenic strains. AB - OBJECTIVE: DA rats are highly susceptible to experimental models of rheumatoid arthritis (RA). Linkage analyses in different models have identified several quantitative trait loci (QTLs) within a 70-cM region of DA rat chromosome 4 (C4). We produced congenic strains for these QTLs in order to map and characterize their impact on arthritis development. METHODS: Selective breeding was used to transfer C4 intervals from arthritis-resistant PVG.1AV1 rats onto DA rats. These congenic strains were evaluated for susceptibility to arthritis induced by intradermal injection of rat type II collagen, pristane (a well-defined synthetic adjuvant oil), mycobacteria, or squalene (an endogenous adjuvant oil used in human vaccine). RESULTS: Rats congenic for PVG.1AV1 genes in the 70-cM region were less susceptible than DA rats to collagen-induced arthritis (CIA), pristane induced arthritis, adjuvant-induced arthritis, and squalene-induced arthritis (SIA). Experiments in subcongenic strains indicated a gene regulating arthritis in males located in a 20-cM interval overlapping the QTL Pia5. A second gene, located in a 10-cM interval harboring the QTL Oia2, attenuated SIA and CIA. The latter caused a change in anticollagen antibody isotype levels toward a pattern similar to that seen in PVG.1AV1 rats. CONCLUSION: The QTL Oia2 regulates arthritis induced both by the nonimmunogenic immunostimulant squalene and by cartilage collagen. In CIA, it also skews anticollagen isotype profiles, suggesting qualitative regulation of autoimmunity. Interestingly, the homologous human chromosome region 12p12-p13 has also been linked to RA, suggesting that genetic and functional dissection of this locus will provide clues to disease pathways that lead to joint inflammation. PMID- 12571865 TI - Geldanamycin inhibits the production of inflammatory cytokines in activated macrophages by reducing the stability and translation of cytokine transcripts. AB - OBJECTIVE: Heat-shock protein 90 (Hsp90) is critical in the intracellular signaling pathways that promote inflammatory cytokine production. Geldanamycin (GD) is a benzoquinone ansamycin that inhibits the function of Hsp90. GD inhibits the production of tumor necrosis factor alpha (TNFalpha) in activated macrophages and suppresses the progression of adjuvant-induced arthritis and experimental allergic encephalomyelitis in rodents. GD has been used to investigate the mechanisms by which Hsp90 regulates inflammatory cytokine production. METHODS: The macrophage cell line RAW264.7 (or primary peritoneal macrophages) was activated with lipopolysaccharide in the absence or presence of GD. The effect of GD on the transcription, stability, and translation of inflammatory cytokine messenger RNA (mRNA) was determined using nuclear run-on assays, mRNA decay assays, and sucrose gradient polysome profiles, respectively. RESULTS: Our data revealed that GD potently inhibits the production of TNFalpha, interleukin-6 (IL 6), and IL-1beta in activated macrophages. Although GD did not significantly reduce the transcription of inflammatory cytokine mRNA, it significantly decreased the stability of these transcripts. Polysome profiles indicated that GD also inhibited the translation of TNFalpha and IL-6 transcripts. These effects may be due, in part, to inhibition of p38 mitogen-activated protein kinase, a kinase known to regulate the stability and translation of inflammatory cytokine transcripts. CONCLUSION: These results indicate that the function of Hsp90 is important in the posttranscriptional control of inflammatory cytokine production. PMID- 12571868 TI - Chronic fatigue syndrome in patients with macrophagic myofasciitis. PMID- 12571867 TI - Association of hospital and surgeon volume of total hip replacement with functional status and satisfaction three years following surgery. AB - OBJECTIVE: To evaluate whether hospital volume and surgeon volume of total hip replacements (THRs) are associated with patient-reported functional status and satisfaction with surgery 3 years postoperatively. METHODS: We performed a population-based cohort study of a stratified random sample of Medicare beneficiaries who underwent elective primary or revision THR in Ohio, Pennsylvania, or Colorado in 1995. The primary outcomes were the self-reported Harris hip score and a validated scale measuring satisfaction with the results of surgery. Both outcomes were assessed 3 years postoperatively. Hospital volume was defined as the aggregate number of elective primary and revision THRs performed on Medicare beneficiaries in the hospital in 1995. High-volume hospitals were defined as those in which >100 such procedures are performed annually, and low volume centers were defined as those in which 12 procedures per year. CONCLUSION: Hospital volume and surgeon volume have little effect on 3-year functional outcome following THR, after adjusting for patient sociodemographic and select clinical characteristics. However, satisfaction with primary THR is greater among patients who underwent surgery in high-volume centers, and satisfaction with revisions is greater among patients whose operations were performed by higher-volume surgeons. Referring clinicians should incorporate these findings into their discussion of referral choices with patients considering THR. Conclusions regarding the effect of volume on longevity of the implants must await longer-term followup studies. Finally, further research is warranted to better understand the association between hospital and surgeon procedure volume and patient satisfaction with surgery. PMID- 12571869 TI - Effect of caffeine consumption on efficacy of methotrexate in rheumatoid arthritis. PMID- 12571870 TI - The case against haste in the treatment of rheumatoid arthritis: comment on the editorial by Pincus et al. PMID- 12571871 TI - An alternative perspective on treatment for diabetes, hypertension, and rheumatoid arthritis: comment on the editorial by Pincus et al. PMID- 12571873 TI - Early and aggressive treatment affects the association of HLA-DRB1 genes with progression of joint damage: comment on the article by Lard et al. PMID- 12571874 TI - Effect of interferon alfa-2b and ribavirin treatment on hepatitis C-associated cryoglobulinemia and rheumatoid factor: comment on the article by Vassilopoulos and Calabrese. PMID- 12571875 TI - Characterization of flares in patients with systemic lupus erythematosus: comment on the article by Ho et al. PMID- 12571876 TI - Epitope spreading in systemic lupus erythematosus: comment on the article by Monneaux and Muller. PMID- 12571878 TI - A six-month randomized, controlled, double-blind, dose-response comparison of intravenous pamidronate (60 mg versus 10 mg) in the treatment of nonsteroidal antiinflammatory drug-refractory ankylosing spondylitis: comment on the article by Maksymowych et al. PMID- 12571881 TI - [A study on the relationship between tree age and bark quality in Magnolia officinalis]. AB - Contents of magnolol and honokiol in 76 samples of Magnolia officinalis collected from 11 counties in Zhejiang, Fujian, Sichuan, Guangxi, Hunan, Jiangxi and Hubei are analyzed by means of HPLC. A study on the effect of tree age on effective ingredients in Magnolia offcinalis shows that the influence of tree age on the content of magnolol and honokiol is correlated with the varieties under cultivation, but a small effect on the content of the phenols with an indent at the tip of leaves. The content of magnolol in other types of leaves increases rapidly with increase in age, diameter of the trunk and thickness of the bark, with not many changes found at the age of 12 or up. Increase in age may be favorable to the full expression of oily characteristic. These results provide a scientific base for the determination of optimal time for harvesting bark from artificially established Magnolia officinalis stands. PMID- 12571882 TI - [Analysis of trace elements and composition of protein in sika blood]. AB - The levels of trace element and protein were determined in eight male sikas. The results revealed that the levels of phosphorus, zinc, copper, ferrin and manganese were 2.68 mmol/L, 51.3 mumol/L, 21.4 mumol/L, 255.6 mumol/L and 0.16 mumol/L respectively and which were significantly higher than the human normal values. Serum levels of albumin and globulin were 34.9 g/L and 35.5 g/L respectively. Among them, the level of gamma-globulin was the highest (25.2%) which was threefold than the human serum normal value. From the above results, sika blood has great value in medicinal use. PMID- 12571883 TI - [Development of healthy beverage from Eucommla ulmoides foliage]. AB - Effective compositions of Eucommla ulmoides foliage, sampled from the south side west section of the Qinling Mountain area, are analyzed qualiatively and quantitatively. It is shown that the foliage contains plenty of mineral elements, seventeen amino acids, proteins, sugars, alkaloids, flavonoids, aucubin, chlorogenic acid, pinoresinol diglucoside, coffeic acid juice and Jingniping glucoside. Based on the analysis and the main material of Eucommla ulmoides foliage, a healthy beverage functioning especially on resisting tire actions is formulated and produced. This work has provided a dependable new idea for further exploitation and utilization of the natural resources. PMID- 12571884 TI - [Pharmacognostic identification of Ampelopsis grossedentata]. AB - Pharmacognostic identification of Ampelopsis grossedentata was carried out by studying its microscopic characteristics, UV spectra and TLC. The results may provide a scientific basis for the comprehensive development and utilization of the drug. PMID- 12571885 TI - [Pharmacognostical identification on the stem and leaf of Hedera nepalensis var. sinensis]. AB - In this paper, pharmacognostical identification of Hedera nepalensis var. sinensis was studied. The character of medicinal materials, histological and powder characteristics and UV absorption for the stem and leaf of H. nepalensis var. sinensis were mainly reported. PMID- 12571886 TI - [Chemical composition of essential oil from leaves of litsea cubeba and its antifungal activities]. AB - The 24 chemical constituents of the essential oil extracted from the leaves of Litsea cubeba have been identified by means of GC-MS technique. Among which, alpha-cis-ocimene(25.11%), 3,7-dimethyl-1,6-octadien-3-ol(16.85%) and n transnerolidol (13.89%) were the principal components. In vitro this oil had a manifest antifungal activities with MIC between 0.03-0.4 microliter/ml for utilized pathogenic fungi and 1.0-2.0 microliters/ml for moulds. PMID- 12571887 TI - [Studies on the chemical constituents of essential oil from roots and rhizomes of Patrinia rupestris]. AB - OBJECTIVE: To study the chemical constituents of the essential oil from the roots and rhizomes of Patrinia rupestris. METHOD: The essential oil was extracted by steam distillation. The components were separated and identified by gas chromatography mass spectrometry and elucidated on the standard mass spectral data. Gas chromatography showed their percentages by area normalization method. RESULTS: Seventeen constituents were identified which make up 74.57% of the oil. CONCLUSION: Transcaryophyllene (38.41%) was the main constituent, and other constituents include alpha-gurjunene, caryophyllene oxide, humulene, 9,12 octadecadienoic acid, etc. PMID- 12571888 TI - [Studies on the anti-enteropathogenic bacteria action of Herba Pogostemonis extracts]. AB - The results of antimicrobial test in vitro of water-extracts and volatile oil of the Herba Pogostemonis shown that the water-extracts posses some inhibitory activity on all the tested bacteria, including Salmonella, Escherichia coli, Shigella flexneri and Staphylococcus aureus, while inhibition to S. aureus is more stronger than to Enterobacteriaceae (P < 0.01). In addition, the activity of anti-enteropathogens of water-extracts of the herb collected from Gaoyao county is more stronger than of the herb collected from Wuchuan county (P < 0.01). PMID- 12571889 TI - [Pharmacological study on asthma-relieving decoction (dingchuan tang) treating asthma]. AB - Asthma-relieving decoction (Dingchuan tang) is a famous prescription of treating asthma, but its pharmacology has been not reported. This experiment showed that Asthma-relieving decoction (Dingchuan tang) can protect experimental asthma that blending of histamine and acetylcholine caused (P < 0.05 [symbol: see text] P < 0.01), can resist external trachea smooth muscle's contracting that histamine caused (P < 0.01), and can promote secretion of phenol red of rat's respiratory tract (P < 0.01). Results indicated that Asthma-relieving decoction (Dingchuan tang) has significant effect of relieving asthma and expelling phlegm. PMID- 12571890 TI - [Study on activities of eight kinds of traditional Chinese medicine against urogential Chlamydia trachomatis]. AB - In order to screen the susceptibilities of urogenital chlamydia trachomatis (CT) to 8 kinds of traditional Chinese medicine by microculture technigue of McCoy cell in Vitro. The results showed that 8 kinds of Chinese medicines all had activities against urogenital CT. Their minimal inhibitory concentrations (MIC) were from 0.49 mg/ml to 15.63 mg/ml. The number and the volume of inclusions were reducing gradually, disappeared finally. These traditional Chinese medicines hadn't any action of cytotoxicity to McCoy cell. The result of this study will be useful for the further study. PMID- 12571891 TI - [Grey relational analysis on early bolting of Angelica and its main factors]. AB - Selecting the principal environmental and cultivated factors from the main production areas of Angelica sinensis in Gansu, the problem of early bolting were analysed with grey system theory. After the quantitative analysis, the relational grade was: elevation > precipitaton > density of crop > organic fertilizer > phosphorus fertilizer > nitrogen fertilizer. The research would provide scientific basis for check on the rate of the early bolting of Angelica effectively as well as improve the product and benefits. PMID- 12571892 TI - [Pharmacognostical identification on Chinese drug migancao]. AB - In this paper, Migancao (Phyllanthus matsumurae Hayata) was identified on the character, microproperties, UV spectra and TLC. It provides scientific basis for its comprehensive development and ultilization. PMID- 12571893 TI - [Identification of Panax quinquefolius, P. ginseng and P. notoginseng by protein fingerprints]. AB - Establish the way of protein fingerprints to identify P. quinquefolius, P. ginseng and P. notoginseng of genus Panax by SDS-PAGE, Tris-tricine and Western blot. The fingerprints of Panax species from 28 KD to 58 KD and 55 KD are in common. Very different fingerprints of P. quinquefolius are found below 28 KD. The fingerprints above 58 KD seemed to be peculiar for P. ginseng. Hence the approach may be used as a means to authenticate the concerned Panax species, especially for identification of P. quinquefolius. PMID- 12571894 TI - [A preliminary study of two Chinese herbs protective tablets on some Chinese traditional medicines]. AB - The protective action of 2 tablets of Chinese herb to 5 Chinese traditional medicines against harm of insects and mildews was tested. It was found that 2 tablets have a obvious effects of insect-repellency and mouthproof in the test with Homalomena occulta and Prunus armeniaca, the bore in the medicinal materials was decreased 94.95% and 95.55% respectively than that of check. The tablets have some effects of mildewproof in the test with Tussilago farfara. PMID- 12571895 TI - [Comparison of protocatechuic aldehyde in different processed products of Salvia miltiorrhiza]. AB - The contents of Protocatechuic aldehyde in crude and four different processed products of Salvia miltiorrhiza were determined by HPLC. The results showed that, after processing, the contents of Protocatechuic aldehyde decreased in varying degrees. Among them, the processed products fried with wine, vinegar and no materials decreased 17.14%, 20.00% and 42.86% respectively. The carbonized product decreased 60.86%. PMID- 12571896 TI - [Studies on the increasing-effect components for molluscicides in nut of Areca catech]. AB - The increasing-effect components for molluscicides isolated from the dry nut of Areca catech L. were studied. The results showed that arecoline was the most effective and it could decrease remarkably the amount of drugs i.e. saponium of Phytolacca acinosa, SPA, and sodium pentachloro phenate, NaPCP, when used together. PMID- 12571897 TI - [Chemical constituents of essential oils from the dried flower buds of Lonicera similis]. AB - The chemical constituents of essential oils from the dried flower buds of Lonicera similis Hemsl. were analyzed by GC/MS for the first time. 94 chemical compounds were identified and the total content of the identified constituents in the essential oils was 88.72%. The percentage of each constituent was given. PMID- 12571898 TI - [Experimental study on antitumor activity of the root of Eurphorbia helioscopia in vivo]. AB - Previous studies showed EWE could strongly inhibit the proliferation of tumor cells in vivo. Experimental results also showed that EWE had antitumor effect in S180-bearing mice and H22-bearing mice and prolonged life-span in S180-bearing mice in this paper, Furthermore, it was found that EWE could improve the immune function of S180-bearing mice. Therefore EWE can be considered as a potent antitumor herb. PMID- 12571899 TI - [Studies of Echinopsis multiplex on antineoplastic effects]. AB - Fresh Echinopsis multiplex decoction(p.o. 30 g/kg and 60 g/kg) could inhibit the growth of solid tumour S180 and Lewis pulmonary carcinoma, prolong the survival date of the mice with the tumours. In vitro, the serum of animal, which took the herb, significantly suppressed the proliferation of EAC tumour cells and inhibited the YAC-1 tumour cell DNA synthesis when compared with the serum of normal animal. The decoction did not reduce the WBC, but increased the number of PFC of spleen cells at the effective dose in mice with tumour, indicating that it did not impair the immune system, but enhance the immune function. PMID- 12571900 TI - [Experiment on polyploid induction of Angelica dahurica var. formosana]. AB - Colchicine solution was applied to the primary adventitious buds of Angelica dahurica var. formosana in vitro to induce the polyploid. Compared with non treated plantlet, the morphology, microhisology, and chromosome number of treated plantlets are varied. It proved that the polyploid induction was effective. PMID- 12571901 TI - [Resources and utilization of anticarciogenic medical fungi]. AB - This paper summarizes the resources of anticarcinogenic medical fungi, and the information of distribution, eco-environment. It outlines the present statuts of the exploitation and utilization of some species of common anticarcinogenic medical fungi such as Ganoderma lucidum, Coriolus versicolor, Polyporus frondosus etc. The anticarcinogenic mechanism of medical fungi is discussed. Suggestions on how to development and utilize rationally these resources are offered as well. PMID- 12571902 TI - [Studies on identification in Sarcandra glabra by microscopic structure and TLC]. AB - Studies on microcopic structure and TLC in Sarcandra glabra (Tunb.) Nakai are reported. Not only does it counteract weaknesses of former research, but it clarifies some matters well. The method to ideutify accurately this drug is provided. PMID- 12571903 TI - [Identification study on pig's hoofnail]. AB - The identification characteristics of pig's hoofnail on properties and tissue slice was initially probed from the viewpoint of identification in this paper in order to make basis for the systematic research on replacing Squama manitis. PMID- 12571904 TI - [Microscopic identification on the Folium Mori and the leaves of its allied species from Shandong]. AB - The Folium Mori and the leaves of its allied species Morus australis Poir., M. mongolica Schneid. from Shandong were identified. The result shows that they are identified easily and accurately according to epidermis, trichomes and blade. PMID- 12571905 TI - [Studies on packing bag and technique for storing up Codonopsis in quality guaranteed]. AB - Under different roots water contents and storing temperature degree, vacuum packing, nitrogen gas packing, carbon dioxide packing were used with different packing bag, to select and study the optimum packing bags and techniques for storing up Codonopsis in quality guaranteed. The results showed that, below the 15% water contents for Codonopsis roots, the two ways of vacuum packing bag with nitrogen gas or carbon dioxide packing and vacuum packing are quite efficient for quality guranteed in Codonopsis. PMID- 12571906 TI - [Gastrointestinal inhibitory effects of sesquiterpene lactones from Atractylodes macrocephala]. AB - Atractylenolide I (8,9-dehydroasterolide, B), 4,15-epoxy-8 beta-hydroxyasterolide (C) and atractylenolide III (8 beta-hydroasterolide, D) from Atractylodes macrocephala Koidz, 28.0, 56.0 mumol/L, inhibited the spontaneous movement of rat isolated ileum with contractile force decreased obviously (P < 0.001), but the rest force not affected (P > 0.05). B, C and D inhibited the spasm enhanced by Ach and His to rat ileum, rightly shifted the dose-response curves and reduced the maximal response (P < 0.05). Pretreatment with B, C or D 28.0, 56.0 mumol/L Ver 0.28 mumol/L significantly antagonized CaCl2-induced contraction of rat isolated ileum. Co bathing of B, C or D 56.0 mumol/L with Atr 0.19 mumol/L weakened the response of ileum to Ach. B, C or D 56.0 mumol/L restrained the spasm of ileum mediated by Neo 2.15 mumol/L. The modified Phenol Red Paste Model demonstrated that B, C or D in dose of 28.0 mg/kg inhibited the gastrointestinal prochoresis of mice significantly. These results suggested that B, C and D inhibit the gastrointestinal movement of rat and the spasm induced by the agonists. Their activities are related to the inhibition of cholinergic system and Ca2+. PMID- 12571907 TI - [Electron microscope observation on effect of kudingcha inspissation tea on small intestine villus in the adiposity rats]. AB - OBJECTIVE: Experimental study on pharmacological action of Guang Dong kudingcha inspissation tea on small intestine villus in the adiposity rats (nutrition obesity). METHODS: By using electron microscope method, check on small intestine villus of 60 experiment rats of just wean and count and analyse and conclude. RESULTS: Under the scan electron imcroscope, the surface configuration on small intestine villus of model group and various kudingcha dosage groups is similar to the blank (P > 0.05), but fenfluramine group appear constriction on top end of small intestine villus. CONCLUSION: Compring with fenfluramine, Guang Dong kudingcha inspissation tea has not effect on configuration of small intestine of adiposity rats (nutrition obesity), but has more strong modulation function on fat tissue lipocyte hypertrophy and quantitative. PMID- 12571908 TI - [Application study of yangkang biological fertilizer on Panax notoginseng cultivation]. AB - Yangkang Biological Fertilizer was sprayed on Panax notoginseng. The result indicated that the yield and the surviving rate can be raised 25.2% and 15.2% respectively. The best spraying practice were: in the unfolding stage of leaves (April) and bloom stage (July), with Yangkang Biological Fertilizer plus water (1:400), at four times every stage. PMID- 12571909 TI - [The study of amino acid constituents on fungus garden of Odontotermes formosanus]. AB - The qualitative and quantitative analysis of twenty free amino acids for fungus garden of Odontotermes formosanus and its aqueous extracts were studied by auto amino acid analytic instrument. PMID- 12571910 TI - [Habitats and present situations of 13 rare precious medicinal materials in Funiu mountains]. AB - The living resources are very abound in the Funiu mountains area because of its special geographical position and climate, and there are also many rare precious living beings and Chinese medicinal herbs, for example, Taxus mairei, Ginkgo biloba, Gastrodia elata, Dendrobium huashanense, Physochlaina infunbularis, Cypripedium japonicum, Dysosma versipellis, Moschus moschiferus, Megalobatrachus davidanus, Succinum, etc, and conscious protection and rational development is an urgent task. PMID- 12571911 TI - [TLC-identification of radix Rehmanniae and its mixed medicinal materials in Pharmacopoeia]. AB - In 237 kinds of medicinal meterials included 71 kinds of Chinese patent medicines composed in Pharmacopoeia such as Liuweidihuang Pills, character identify constituent of Radix Rehmanniae was found by TLC, and was successed to use to identification of Radix Rehmanniae in 10 kinds of Chinese patent medicines such as Liuweidihuang Pills. PMID- 12571912 TI - [Studies of volatile oils from the flowers of Chrysanthemum nankingense and Chrysanthemum indicum]. AB - Chemical constituents of volatile oils from the flowers of Chrysanthemum nankingense and C. indicum were studied by GC-MS. 59, 30 components were identified respectively. PMID- 12571913 TI - [A primary study on distribution of macro and trace elements in water extracts of the root and semen of Ziziphi spinosae]. AB - A determination of several macro and trace elements in the water extracts of the root and semen of Ziziphi spinosae is presented. The result implies that the distribution of different elements in the water extract is different and it has supplied reference data for the further studies of state analysis of above elements. PMID- 12571914 TI - [Effects of the centipede aqueous extract on animal gastrointestinal function]. AB - In order to investigate the mechanism of Centipede on the function of animal gastrointestinal digestion, we studied the effects of cetipede aqueous extract on animal digestion, secretion and intestional movement. The results showed that the Centipede extract increased the volume of gastric juice and acid, activity of pepsin, volume of pancreatic juice and protein in rat. The Centipede extract also accelerated the mice intestinal movement. The above results revevaled that Centipede could regulate the function of the gastrointestinal digestion, secretion and movement. PMID- 12571915 TI - [Supercritical-CO2 fluid extraction of fatty oils from Perilla frutescens seeds and pharmacological study]. AB - Fatty oils from the seeds of Perilla frutescens were extracted by supercritical CO2 fluid or petroleum ether and its quality was analyzed by GC-MS method. Pharmacological action of lowering blood-lipid for the oils was studied. The results showed that the technology of supercritical-CO2 extraction of the oils of P. frutescens seeds is superior than that of petroleum ether and the oil from SFE CO2 have the action of lowering blood-lipid with low toxicity. PMID- 12571916 TI - [Experimental study on the effect of nourishing yin and tranquilizing of yangyin anshen koufuye]. AB - Yangyin Anshen Koufuye could obviously decrease the spontaneous activity of rats, effectively shorten the process of falling into sleep of normal rats which were injected pentobarbital sodium and prolong the sleeping time. The effect of sedation and hypnotism were the same as Zhaoren Anshen Koufuye. Yangyin Anshen Koufuye also could markedly reduce the serum tensity of T3, T4 of hyperthyroid rats, which equaled to TCM's deficiency of yin. It also could prevent the glycogen content of liver from decreasing, decrease heart rate, resist weight losing, thus to show the effect of nourishing Yin and tranquilizing. PMID- 12571917 TI - [Comparison of the components of Ganoderma lucidum and Ganoderma japonicum]. AB - By means of spectrophotometric methods, the contents of polysaccharides about Ganoderma lucidum (including mycelia) and Ganoderma japonicum are determined, HPLC and TLC methods for idntification of their triterpenic acids are given. PMID- 12571918 TI - [Analysis of ethereal extracts from Odondotermes formosanus and fungus garden]. AB - Nine, twelve compounds were isolated and identifacted from ethereal extracts of Odotntotermes formosanus (insect body) and its fungus garden respectively, by GC MS analysis. PMID- 12571919 TI - [Character identification of 12 kinds of pilose antler medicinal materials]. AB - In this paper, commercial medicinal materials of 12 kinds of pilose antler, Cervus nippon, C. elaphus, C. albirostris, C. unicolor, C. eldihainanus, C. timorensis C. porinus, Dama dama, Rangifer farandus, Alces alces, Elaphurus davidianus, Capreolus capreolus were compared and identified. A key and simple character illustration were listed. PMID- 12571920 TI - [Studies on the chemical constituents of Adina rubella (II)]. AB - Three constituents were isolated from Adina rubella. Their structures were identified as beta-sitosterol, quinovic acid, 3-oxo-urs-12-ene-27, 28-dioic acid. PMID- 12571921 TI - [Study on carbazole alkaloids of Murraya microphylla]. AB - Four carbazole alkaloids were isolated from the methanol extract of Murraya microphylla. Their structures were identified by spectral analysis and chemical evidence, which were identified as koenigine, bis-6-hydroxy-7-methoxygirinimbine, girinimbine and mukonicine. All the compounds were discovered from this plant for the first time, and bis-6-hydroxy-7-methoxygirinimbine was a new compound. PMID- 12571922 TI - [Effects of the water-soluble extracts from the single herb of ganduqing against hepatitis B virus in vitro]. AB - Comparing with Ara-Amp, the effects of the water-soluble extracts from the single herb of the formula for Ganduqing on HBeAg and HBsAg expression in 2.2.15 cells were studied. The results showed that the extracts of Serissa serissoides (DC) Druce, Hibiseus mutabilis Linn, Paedeuia scangens (Lour) Merr var tomentosa (BL) Hand-Mazz, Plumbago zeylanica L, Garcinia oblougifolia Champ and Begpnia edulia Levl had marked inhibition effects on HBeAg and HBsAg which expressed by 2.2.15 cells. PMID- 12571923 TI - [Experimental study on pharmacological action of fukean tablet on gastrointestinal tract]. AB - The pharmacological effects of FuKean Tablet(FKT) on gastrointestinal tract were studied. The results showed FKT cured the diarrhea induced by rhubarb or senna, prominently reduced the quantity of feces and delayed the time of excreting charcoal powder in the diarrhea induced by rhubarb or senna model NIH mice; FKT inhibited intestinal propelling-movement in diarrhea induced by senna mmodel NIH mice. FKT inhibited the automatic contraction and Ach induced spasmodic contraction of isolated rabbit intestine(decreased the frequency and intensity of contraction). The above results revealed that the pharmacological base of curing the urgent disease in gastrointestinal tract may be curing the diarrhea, inhibiting intestinal propelling-movement and inhibiting the contraction of intestine. PMID- 12571924 TI - [Acute toxicity of Stephania cepharantha]. AB - The acute toxicity of Stephania cepharantha was studied, LD50 of aqueous extract of its wet and dry root tuber oral were 41.4 g/kg and 22.9 g/kg respectively. PMID- 12571925 TI - [Investigation on preparation procedure for compound shenglin oral]. AB - The optimum preparation procedure for compound Shenglin oral was studied by orthogonal test (including 5 factors: water-adding amount, decocting time, decocting times, concentrating scale, alcohol-precipitation concentration with 3 levels every factor). As a result, A1B1C3D1E1 is the procedure of choice: 24 times of water-adding amount, decocting 3 times, 1.5 hours of total decocting time, 0.8 g/ml of concentrating scale and 36% of alcohol-precipitation concentration. PMID- 12571926 TI - [Studies on polysaccharides from Holothuria leucospilota]. AB - A powder was obtained from the dried body of Holothuria leucosilota (Brandt). According to IR and UV spectra, paper chromatography and the physical and chemical data, it is showed a sulfated mucopoly-saccharide consisted of galactosamine, glucouronic acid, fucose and sulfate with the molar ratio of 1:0.96:0.78:1.98 respectively. The specific rotation, intrinsic viscoslty and molecular weight of this polysaccharide were all presented. PMID- 12571927 TI - [Chemical pattern recognition of traditional Chinese medicine xixin (II)]. AB - The data matrix of macro and trace elements from 45 samples of traditional Chinese medicine Xixin were treated with principal component analysis and cluster analysis in order to establish why and how Xixin samples from different species can be recognized. According to the results of hierarchical clustering, three species of Xixin (Asarum heterotropoides Fr. var. mandshuricum (Maim.) Kitag., Asarum sieboldii Miq. var. seoulense Nakai and Asarum sieboldii Miq.) were classified and recognized accurately. PMID- 12571928 TI - [Assay of essential oil from Evodia rutaecarpa var. officinalis by GC/MS]. AB - In the essential oil from Evodia rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang, 55 constituents were analysed by means of GC/MS/DS. The contents of these compounds are 92.0% in the essential oil from this plant. The main compounds are terpenoids. PMID- 12571929 TI - [Studies on chemical constituents of the stem bark of Taxus cuspidata]. AB - In order to study the active constituents in the stem barks of Taxus cuspidata Sieb. et Zucc., led to the isolation and structural determination of three compounds. On the basis of chemical evidences and spectral analysis (UV, IR, 1HNMR, 13CNMR and FAB-MS,) the structures of I, II and III were elucidated as baccatin I 1-hydroxy, taxinine J 2-deacetoxy and beta-sitosterol. Compound I was found from this plant for the first time. PMID- 12571930 TI - [Effect of total diterpene of Rabdosia macrocalyx on mice of normal and immunosuppression]. AB - OBJECTIVE: The regulatory effects of total diterpene of Rabdosia macrocalyx (Dunn) Hara on immune function were studied. METHODS: The experimental immunosuppressive mouse models were induced by injection of cyclophosphamide. RESULTS: The index of thymus gland and spleen in normal and immunosuppressive mice were increased significantly. So were the level of hemolysin in serum when 50 mg/kg total diterpene were given. Delayed hypersensivity were also enhanced at the same condition. But there didn't show any change in carbon clearance test. CONCLUSION: Total diterpene of Rabdosia macrocalyx (Dunn) Hara possesses obvious regulatory action on immune function in mice. PMID- 12571931 TI - [The antitumour activity of total saponin of Clematis chinensis]. AB - This paper has shown the cytocidal effects of total saponin of Clermatis chisensis Osbeck (CCS) on experimental tumours EAC cells, S180A cells and HepA cells in vitro, using microculture method 72 hours. The IC50 were 242 micrograms/ml, 193 micrograms/ml and 130 micrograms/ml respectively. In vivo the growth of transplanted mouse tumour(S180) was inhibited by the CCS with inhibitory rates 40.3% (0.25 g/kg), 55.1% (0.5 g/kg), 53.0% (1 g/kg), but CCS(0.2 g/kg, 0.4g/kg, 0.5 g/kg) hasn't effect on survival time of tumour mice(S180A). PMID- 12571932 TI - [Studies on extracting procedure for huoxue jiangzhi capsules by orthogonal analysis]. AB - The contents of tanshinone IIA, protocatechuic aldehyde and anthraquinone were used to evaluate the extracting procedure for Huoxue Jingzhi capsules by orthogonal analysis. The best one can be described as: the drug powder is reflowed 2 times, 3 hours each, percolated by 80% alcohol with the 4 times weight of powder. PMID- 12571933 TI - [Discussions with the author of Materia Medica Textual Research on the original plants of the medicinal herbs in Ardisia (comment)]. AB - Objections about the conclusions that six plants of eleven medicinal herbs given by the Materia Medica Textual Research on the Original Plants of the Medicinal Herbs in Ardisia were raised in this paper. It is considered that the eleven herbs recorded in ancient Chinese materia medica books are at least belonging to fourteen species in Ardisia and one species in Solamum. PMID- 12571934 TI - [Efficacy of ablendazole emulsion in treatment of 212 patients with cystic echinococcosis]. AB - OBJECTIVE: To verify the efficacy of abendazole emulsion, a new formulation of abendazole, in treatment of human cystic echinococcosis. METHODS: 212 patients with liver cystic echinococcosis were treated orally with albendazole emulsion at a daily dose of 10 mg/kg or 12.5 mg/kg for 3 to 12 months or over one year. The therapeutic efficacy was mainly evaluated by image feature examined with B ultrasound examination, a short-term efficacy at the completion of treatment and a long-term efficacy followed-up for 1-4 years. RESULTS: In 212 patients treated with albendazole emulsion at a daily dose of 10 mg/kg and 12.5 mg/kg, the average cure rate, improved rate and the rate of no avail were 74.5%, 99.1% and 0.9% respectively after termination of the treatment, and the average long-term rates were 83.1%, 89.3% and 0.6% respectively. Recurrence occurred in 18 patients(10.2%). The results indicated that the best efficacy was seen in patients treated with albendazole 12.5 mg/kg daily for 9 months. Better response was also found when the recurrent patients were re-treated with albendazole. CONCLUSION: The efficacy of albendazole emulsion on patients with liver cystic echinococcosis is much better than that of albendazole tablet or capsule and mebendazole. Meanwhile, the efficacy of albendazole emulsion is stable with less adverse effects. The results suggest that albendazole emulsion could be the drug of choice for treatment of cystic hydatid disease. PMID- 12571935 TI - [Gene cloning, construction and expression of single-chain Fv (scFv) against the membrane protein of Schistosoma japonicum]. AB - OBJECTIVE: To construct single chain antibody specific to membrane protein of Schistosoma japonicum by genetic engineering technique. METHODS: The VH (heavy chain variable region) and VL(light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using VH and VL genes, then expressed by IPTG. RESULTS: The VH and VL genes were obtained through PCR. The DNA sequences showed that VH and VL were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin fused proteins about 36.2 kDa. CONCLUSION: A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed. PMID- 12571936 TI - [Two cases of ocular myiasis]. PMID- 12571937 TI - [Efficacy of albendazole immunoliposome against echinococcosis granulosus in mice]. AB - OBJECTIVE: To evaluate the effect of albendazole immunoliposome (IL-Alb) against Echinococcus granulosus. METHODS: Mice infected with protoscolices of E. granulosus were divided into five groups. Four groups were treated with albendazole (Alb), albendazole liposome (L-Alb), albendazole sulfoxide liposome (L-Albso), and IL-Alb respectively at a dosage of 100 mg (Alb)/(kg.d) x 5 d for 3 courses. The fifth group was established as control. The major criteria for evaluating the effects included a reduction rate of E. granulosus tissue wet weight, histopathological examination of the cysts by both light microscopy and electron-microscopy, and the content of albendazole-sulfoxide in cysts detected by HPLC. RESULTS: The efficacy of albendazole immunoliposome was significantly higher than that of albendazole liposome, and much higher than that of albendazole. The reduction rates of cyst tissue weight of IL-Alb group, L-Alb group and Alb group were 91.5%, 80.3%, 61.2% respectively as compared to control group; the concentration of Albso in cyst tissue of the above groups were 5.15 micrograms/g, 2.18 micrograms/g, 0.76 microgram/g respectively (P < 0.01). The histopathological damages of cysts were also found most severely in the group of IL-Alb. CONCLUSION: Immunoliposome as a targeting carrier may significantly strengthen the therapeutic effect of albendazole on echinococcosis granulosus. PMID- 12571938 TI - [Second sampled survey on the distribution of human parasites in Zhejiang Province]. AB - OBJECTIVE: Twenty-six species of human parasites were recorded in Zhejiang Province in 1987-1989 as a part of the national investigation on the distribution of human parasites, and the total prevalence was 80.2%. In order to find out possible changes on the composition of parasite species and decrease of prevalence after control intervention particularly mass chemotherapy in the past years and provide evidence for an improved control strategy, the second sampled survey was carried out from 1998 to 1999. METHODS: Ten counties were identified randomly out of 28 counties where the last survey was conducted following the same sampling method. Same technics were used for case detection and data processing. RESULTS: The total infection rate in a sample of 15,698 was 22.84% was 22.84% in 30 investigation spots in 10 counties, and 17 species of parasites were revealed. The overall prevalence was reduced by 71.51% in comparison to that of 1989, and the number of parasite species was 17, 9 less than that of the last investigation. CONCLUSIONS: The prevalence of human parasites has greatly declined in this province due to the socioeconomic development and adoption of comprehensive control measures focusing on mass chemotherapy in the past decade. PMID- 12571939 TI - [Partial sequence of sporogony stage-specific 18S ribosomal DNA of Plasmodium yoelii and its application for detection of parasites]. AB - OBJECTIVE: To determine sequence of sporogony stage-specific (S type) 18S ribosomal RNA gene of Plasmodium yoelii (P. y) By265 strain, and by using it to detect the malaria parasites within vector mosquito. METHODS: A pair of conserved DNA primers, universe primer (Pu) and reverse transcription one (Pr), was designed and synthesized according to sequence of the 18S rRNA gene of Plasmodium berghei (P. b). The segment of the S type 18S rDNA of P. y was amplified by reverse transcript-polymerase chain reaction (RT-PCR) from dissected midguts of Anopheles stephensi infected with P. y on the 7th day after infective blood-meal, and its sequence was then determined. One P. y sporogony stage-specific primer (Pys) was selected according to the sequence. Using this primer and Pr, the parasites within mosquitoes were semi-quantitatively detected through RT-PCR between 1-7 d post-infection. RESULTS: The length of the amplified segment was 920 bp. Alignment in match region of the 18S rDNA among S type of P. y (PyS), S type of P. b (PbS) and asexual blood stage-specific one of P. y (PyA) revealed that the similarity between the former and the latter two reached 95.3% and 94.0% respectively. The density of amplified band was significantly concordance with the intensity of oocyst in the midgut. Sensitivity of RT-PCR method was higher than that of the traditional dissection and oocyst observation also. The assay could detect the 18S rRNA molecule of the parasites on the third day post infection while their oocysts were difficult to be recognized under an optical microscope at that time. CONCLUSION: This S type 18S rDNA sequence in P. y species was first reported (AF266261). As a molecular marker, it could be applied to monitoring the parasite development in its vector at an earlier stage semi quantitatively with an adequate sensitivity and specificity. PMID- 12571940 TI - [Screening and cloning of genes encoding Schistosoma japonicum antigens related to the serum antibodies in Mirotus fortis]. AB - OBJECTIVE: To understand and identify the molecules related to the natural resistance to Schistosoma japonicum infection in Mirotus fortis. METHODS: Sera from Mirotus fortis without schistosome infection were collected. The S. japonicum adult worm cDNA library was immunologically screened with the sera. The positive recombinants were identified, cloned, sequenced and analysed with software and internet. RESULTS: Seven genes encoding antigens relevant to sera antibodies in Mirotus fortis were cloned and sequenced. These antigens included glyceraldehyde 3-phosphate dehydrogenase (GAPDH), serine protease inhibitors (SERPIN), 70 kDa heat shock protein (HSP70), 22.6 kDa membrane-associated antigen, paramyosin (Sj97), cytochrome C and cathepsin B. CONCLUSION: Many protein molecules might have been involved in natural resistance to S. japonicum infection in Mirotus fortis. The above 7 kinds of molecules may be identified as new candidates of vaccine against S. japonicum infection. PMID- 12571941 TI - [Cloning and identification of an unknown gene encoding 10.6 kDa protein of Schistosoma japonicum]. AB - OBJECTIVE: To screen a new schistosome vaccine candidate. METHODS: Schistosoma japonicum adult cDNA library was screened using sera from immune rabbits vaccinated with irradiated cercariae and monoclonal antibodies against membrane antigen of S. japonicum schistosomula. Three different fragments of S. japonicum cDNA genes were cloned into pGEM-T vector. The sequences of the inserts were determined using an automatic DNA sequencer and were analysed using Blast program. One of the unknown genes (B8) was selected and its ORF sequence (291 bp) was subcloned into eukaryotic expression vector. The recombinant plasmids were identified by restrictive enzymes and PCR amplification. The positive recombinant plasmids (pBK/SjB8) were transformed into host bacteria XL1-blue, and were then induced by IPTG for expression. SDS-PAGE and Western blotting analysis of total cellular protein from the bacteria were performed to detect the gene products. RESULTS: The results demonstrated that ORF of SjB8 gene was subcloned into the plasmid pBK-CMV and could express as fusion protein in XL1-blue. The results of SDS-PAGE and Western-blot also showed that the molecular weight of the fusion protein with 3 kDa beta-galactosidase was approximately 13.6 kDa and the actual molecular weights of the SjB8 was 10.6 kDa. The expressed fusion product of pBK/Sj-B8 could be recognized by immune serum and McAb. CONCLUSION: A new gene of S. japonicum vaccine candidate (SjB8) was cloned into eukaryotic expression vector pBK-CMV and could express 10.6 kDa schistosome protein. The results provide foundation for further study of the protein for its possibility as candidate vaccine. PMID- 12571942 TI - [Effect of dibutyl phthalate on demodicidosis]. AB - OBJECTIVE: To evaluate the curative effect and safety of dibutyl phthalate on demodicidosis. METHODS: A single blinded and controlled study of human demodicidosis treated with dibutyl phthalate was conducted. One hundred and forty three patients with demodicidosis, including 81 acne and 62 rosacea, randomly divided into trial and control groups. The trial group was treated with dibutyl phthalate and control group with "new fumanling" cream twice a day in the early morning and evening respectively for six weeks consecutively. RESULTS: The rates of excellent, good, and fair efficacy and total effective rate in the trial group with acne were 53.7%, 41.5%, 4.9% and 100% respectively, with a significant difference to the control group (P < 0.05). The rates in the trial group with rosacea were 40.6%, 40.6%, 18.8% and 100% respectively, with no statistical difference to the control group (P > 0.05). No complaint of side effects in the trial group was recorded. CONCLUSION: Dibutyl phthalate is highly effective to demodicidosis without prominent adverse reactions. PMID- 12571943 TI - [Preliminary study on isolation, purification and hydrolytic activity of cysteine proteinases in Entamoeba histolytica]. AB - OBJECTIVE: To explore the invading mechanism of amebae in lamina porpria and observe the interaction between the cysteine proteinase (CP) of Entamoeba histolytica and laminin. METHODS: CP was identified by laminin-sepharose affinity chromatography, followed by isolation, purification and inhibitor experiment. The hydrolytic activity was measured by gelatin electrophoresis. RESULTS: Purified CP of E. histolytica showed a strong affinity with laminin. The molecular weight of CP is 27 kDa. It can be inhibited by EC-64 and exhibited a protein hydrolytic activity. CONCLUSION: The specific affinity and hydrolytic activity of CP might play an important role in its invasion to the basement membrane of intestinal mucosa. PMID- 12571944 TI - [Re-utilization of parasite eggs in specimens for teaching purpose]. PMID- 12571945 TI - [Studies on immunological reaction of the antiserum of recombinant secreted protein from Ancylostoma caninum]. AB - OBJECTIVE: To analyze the immunological reaction of the antiserum of recombinant secreted protein from Ancylostoma caninum with antigens of various species hookworms at different developmental stages. METHODS: SDS-PAGE and ELIB technique were employed in the study. RESULTS AND CONCLUSION: The protein component of Ac rAsp-1 was 45 kDa, its immune serum can recognize the antigens of Ac-L3 and Ac rAsp-1 protein, but not react to the antigens of Ad-A, Ad-L3, Na-A, Ac-A, Nb-A and Ac-rAsp-2 protein. The protein component of Ac-rAsp-2 was 24 kDa, its immune serum can recognize the antigens of Ad-A, Ad-L3, Na-A, Ac-A, Ac-L3 and Ac-rAsp-2 protein, but not react to the antigens of Nb-A and Ac-rAsp-1 protein. PMID- 12571946 TI - [Morphological and ultrastructural observation of Blastocystis hominis]. AB - OBJECTIVE: To observe the morphology and ultrastructure of Blastocystis hominis. METHODS: Morphological observation was made with 4-5 days cultured B. hominis by light microscopy, and similar material fixed with 4% glutaraldehyde was used for transmission electron microscopy. RESULTS: Several forms of B. hominis were observed including vacuolar, granular, amebic, multifission and cystic forms. The multiplication patterns of B. hominis included both binary fission and sporogony. Under transmission electron microscope, the nuclei, mitochondria, rough endoplasmic reticula and lysomes were observed in addition to lipid droplets in its cytoplasm, and glycogen in the central vacuole. CONCLUSION: The central vacuole of vacuolar form may be related to the storage of the excreta. The amebic form of B. hominis might be pathogenic. PMID- 12571947 TI - [Detection of DNA of Toxoplasma gondii in rat by using polymerase chain reaction]. AB - OBJECTIVE: To study the significance of DNA of Toxoplasma gondii in peripheral blood. METHODS: DNA of T. gondii in peripheral blood of 50 infected rats was detected by polymerase chain reaction. A pair of primers was designed, according to the sequence P30 gene specific to T. gondii, to amplify DNA from T. gondii by PCR. RESULTS: The primers amplified DNA specifically from T. gondii and could not amplify DNA from humans, uninfected rat and mouse and from Trichomonas vaginalis and Entamoeba histolytica. DNA of two Toxoplasma parasites was detected by 35 cycles of amplification, indicating a fair sensitivity of the PCR system. CONCLUSION: PCR may have a value for early diagnosis of T. gondii infection in rat. PMID- 12571948 TI - [Ultrasonographic diagnosis of 51 cases with portal hypertension of late stage schistosomiasis]. PMID- 12571949 TI - [Analysis on karyotypes of ANKA strain of Plasmodium berghei]. AB - OBJECTIVE: To analyze the molecular karyotypes of ANKA strain of Plasmodium berghei and demonstrate the size and number of chromosomes. METHODS: To isolate the genome DNA of P. berghei ANKA strain and analyze molecular karyotypes through CHEF-III pulsed field gel electrophoresis (PFGE). RESULTS: The number of chromosomes was found to be 14, and their size ranged from 0.6 Mb to 3 Mb. Chromosomes number 5 to 7 and 9 to 12 appeared co-migrated in the gel. CONCLUSION: PFGE technique is useful for analyzing the molecular karyotypes and may be also useful for further study to locate the special gene on chromosomes and carry out the genetic characters and mechanism of drug resistance. PMID- 12571950 TI - [A case of Parachordodes spp. infection in urethra]. PMID- 12571952 TI - [A case of tertian malaria]. PMID- 12571951 TI - [Observation on eggs of Oncomelania hupensis hupensis with scanning electron microscope]. AB - OBJECTIVE: To observe the structure of the mud hull packed Oncomelania eggs and the surface structure of colloid membrane called the third grade membrane of eggs. METHODS: Scanning electron microscopy was used to observe Oncomelania snail eggs with integral mud hull collected from eastern Dongting Lake. RESULTS AND CONCLUSION: The mud hull of eggs was made of unshapen small humification combined with earth granules with a diameter of 2.6-9.2 microns. The mud hull in 60 um thickness was honeycomb-like in shape with many small holes and small folds on the wall. There were many round or irregularly round hollownesses on the inner layer of mud hull that contacts colloid membrane but no hole through mud hull. There were some protein fiber networks covering on the colloid membrane and apophysis. The structure of the mud hull showed that the exchange of matter was maintained between eggs and outside, and the mud hull is of great importance to regulating temperature and moisture for the growth of eggs by preventing hydrosoluble substances from penetrating into eggs. The protein fiber networks act on gluing mud hull and buffering outside power. The dense glue membrane might be a main barricade to prevent pharmaceutical molecules from penetrating into eggs. PMID- 12571953 TI - [The antimalarial mechanism of artemisinin and its derivatives]. PMID- 12571954 TI - [48 years malaria control in Xuchang City]. PMID- 12571956 TI - [Pinworm infection in primary school students in Luoyang City]. PMID- 12571955 TI - [Curative effect of combined treatments with albendazole and surgery in 1518 cases of cystic echinococcosis]. PMID- 12571957 TI - [Schistosomiasis surveillance in Huzhou City after interruption of transmission]. PMID- 12571958 TI - [Three cases of intraspinal cysticercosis]. PMID- 12571959 TI - [Misdiagnosis of intestinal ascariasis]. PMID- 12571960 TI - [A longitudinal surveillance of schistosomiasis in Tongling county from 1996 to 2000]. PMID- 12571961 TI - [Experience in the treatment of 62 cases of liver alveolar hydatid disease]. PMID- 12571962 TI - [Cloning and identification of deltamethrin-resistance or susceptibility associated genes of Culex pipiens pallens]. AB - OBJECTIVE: To obtain deltamethrin-resistance or susceptibility associated genes of Culex pipiens pallens. METHODS: The differentially expressed genes were obtained by suppression subtractive hybridization (SSH), and identified by cDNA microarray and reverse Northern blotting. RESULTS: 523 and 286 clones were selected respectively in the two directional SSH. 155 and 42 genes were respectively expressed 2-3 and > 3 times higher in the insecticide-resistant strain than in the susceptible strain; 15 and 9 genes were respectively expressed 2-3 and > 3 times higher in the susceptible strain than in the resistant strain. There were 2 genes only expressed in the insecticide-resistant strain. 51 three times differentially expressed clones and 2 specially expressed clones were sequenced. 44 sequences were obtained which belong to 13 new genes. There were 8 over-expressed genes in resistant strain, 7 of which were similar respectively to mitochondrion rRNA gene, 60S ribosomal protein gene, 40S ribosomal protein S4 gene, trypsin gene, chymotrypsin A gene, ospin gene, and 16S ribosomal RNA gene. There were 5 over-expressed genes in susceptible strain, 2 of them being similar with 40S ribosomal protein S29 gene and myosin regulatory light chain 2 gene. In addition, 2 genes specially expressed in resistant strain were similar respectively to glycogen branching enzyme gene and ribosomal protein 46 gene. CONCLUSION: The differentially expressed genes may be associated with deltamethrin-resistance or susceptibility of Culex pipiens pallens. PMID- 12571963 TI - [Induced expression of the variable region of AMA-1 from Plasmodium falciparum]. AB - OBJECTIVE: To express the variable region of AMA-1 gene from Plasmodium falciparum in Escherichia coli. METHODS: Genomic DNA of FCC1/HN was used as template and the variable region of AMA-1 gene was amplified by polymerase chain reaction(PCR). The PCR products were digested by endonuclease BamH I and Hind III, cloned into pBlu2KSP. The nucleotide sequences of the variable region of AMA 1 gene were determined by sequencing. The AMA-1 gene fragment was subcloned into plasmid pQE, expressed in E. coli and induced by IPTG. The fusion product as identified by SDS-PAGE gel electrophoresis and Western blotting were proceeded with anti-AMA-1 sera from rabbit. RESULTS: The size of the variable region of AMA 1 gene from FCC1/HN was 506 bp and encoded 168 amino acids. On SDS-PAGE gel dyed with Coomassie brilliant blue R250, no specific protein band can be discerned, but Western blotting proceeded with anti-AMA-1 sera from rabbit demonstrated that the specific protein band was about 23.0 kDa. CONCLUSION: The variable region of AMA-1 gene from FCC1/HN was able to be expressed in E. coli and analysis of Western blotting demonstrated that the AMA-1 fussion protein contained specific antigenic epitopes. PMID- 12571964 TI - [Study on molecular phylogeny of Schistosoma sinensium based on nuclear ribosomal DNA]. AB - OBJECTIVE: To determine the phylogenetic relationships between Schistosoma sinensium and other Schistosomatid species using DNA sequence data. Two segments of the nuclear rDNA repeat, the second internal spacer (ITS2) and large subunit (LSU/12S) were selected for sequencing. METHODS: Adult worms stored in 100% methanol were washed 3 times with 0.1 x TE (pH8.0) and the genomic DNA was extracted by the GNT-K method. The target regions were amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid pT-adv (Clontech). Recombinant plasmids were amplified in E. coli (strain TOP10), extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of related schistosomes were retrieved from GenBank and aligned with our data in the sequence editor ESEE. Gene trees were constructed in PHYLIP (Version 3.6 alpha July, 2,000) and MEGA (version 2.0 beta build 3) using both Maximum Parsimony and Neighbor-Joining methods. For parsimony analysis, all characters were treated as unordered and with equal weights. At least 3,000 cycles of bootstrapping were carried out. For analysis in MEGA, all gap columns were deleted. Schistosomatium douthitti and Trichobilharzia were used as outgroups. RESULTS: The ITS2 and LSU sequences of Schistosoma sinensium were obtained. The ITS2 sequence of Trichobilharzia sp. was reported here for the first time. CONCLUSION: The phylogenetic trees from these data of nuclear rDNA suggested that S. sinensium belongs to the Asian schistosome group. And this species might be an ancient member in the Asian clade. PMID- 12571965 TI - [Three caseS of subcutaneous myiasis]. PMID- 12571966 TI - [Construction of multivalent DNA vaccine against Schistosoma japonicum]. AB - OBJECTIVE: To construct a multivalent DNA vaccine. METHODS: The multivalent DNA vaccine candidates pBK-Sj26-Sj23, pBK-Sj32-Sj23 were constructed based on the plasmids pBluescript-Sj26, pBluescript-Sj32 and pBluescript-Sj23 with three pairs of specific primers using DNA recombinant technique. In the primers, a synthetic linker sequence encoding a peptide was designed, and the antigen genes Sj26 and Sj23, Sj32 and Sj23 were then ligated. After identification, the quadriceps muscle of mice were immunized with the multivalent antigen genes. Four weeks after immunization, the multivalent antigen genes were present in the muscular tissue of mice by PCR. RESULTS: The eukaryotic plasmids including multivalent antigens of S. japonicum were constructed successfully, and the plasmids including multivalent antigen gene could be stably existing in the muscle tissue of mice and the multivalent antigens could be expressed in the muscle tissue cells of mice. CONCLUSION: A multivalent S. japonicum DNA vaccine has been established. PMID- 12571967 TI - [Aedes albopictus: cloning and identification of the acetylcholinesterase gene fragment from the mosquito]. AB - OBJECTIVE: To isolate, clone and identify the acetylcholinesterase (AChE) fragment from the mosquito, Aedes albopictus, in relation to exploring mechanism of insectide resistance. METHODS: The genome DNA extracted from the mosquito was used for degenerate polymerase chain reaction (PCR) and the two pairs of oligonucleotides encoding the highly conserved protein sequences were used as primers. The reaction products were cloned to T-vector and transfected into E. coli JM 109. The replicative form DNA of recombinant vector extracted from E. coli JM 109 through alkalilysis was identified by the methods of digestion with EcoR I and Sal I and PCR. RESULTS: The products of degenerate primers polymerase chain reaction were obtained and the identified clone belongs to the AChE fragment of the mosquito. CONCLUSION: The clone was identified as the AChE fragment of Aedes albopictus. PMID- 12571968 TI - [Antigen analysis of Trichomonas vaginalis trophozoite by SDS-PAGE and two dimensional gel electrophoresis]. AB - OBJECTIVE: To analyze soluble antigens of Trichomonas vaginalis. METHODS: Soluble antigens of the parasite from a patient suffering from trichomonad vaginitis were analyzed by SDS-PAGE, immunoblotting and two-dimensional gel electrophoresis. RESULTS: A total of 26 distinct protein bands were demonstrated by using 10% resolution gel. Nine of them were main bands, eight with MWs 15-62 kDa, one with MW 97 kDa. By immunoblotting the specific anti-T. vaginalis antibodies raised in rabbit recognized 24 protein bands with 8 main bands in them. Two-dimensional gel electrophoresis revealed up to 43 individual trichomonad polypeptide spots, among which, 9 were main ones. The pI and MWs of these spots were 3.65-5.84 and 27- >100 kDa respectively. CONCLUSION: Eight protein bands out of 26 soluble antigen bands of the parasite showed high immunogenicity. There were 9 main polypeptide spots in 43 polypeptide spots of the parasite. PMID- 12571969 TI - Leishmania mexicana: the circular DNA 1 (CD1) element contains genes encoding nucleotide-binding protein. AB - OBJECTIVE: To determine the nucleotide sequence of cloned CD1 fragments from Leishmania mexicana and find ORFs predicted to have protein coding function. METHODS: CD1 element was separated by CHEF and recovered by agarase, and the digested CD1 fragments were cloned into pZero vector. Nucleotide sequences were determined by the dideoxy chain termination method with the automatic sequencing system ALF using the M13 universal primers. Sequences were analyzed using GCG PCGENE computer programs. RESULTS: The sequence with 4,385 nucleotides was determined and two ORFs were considered to have protein coding function (encoding nucleotide-binding protein). CONCLUSION: Genes encoding nucleotide-binding protein were identified from the amplified CD1 element of Leishmania mexicana. PMID- 12571971 TI - [Study on schistosomiasis control strategy in Ertan reservoir]. AB - OBJECTIVE: To explore means and strategies of preventing the spread of schistosomasis transmission due to the building of Ertan Dam. METHODS: To eliminate the infection sources and Oncomelania snails. To install concrete irrigation and piping system of water supply. To encourage the immigrants to build methane-generating tanks and improve sanitary facilities and conditions for families who live near the water-retaining line. RESULTS: 2,360 people and 152 cattle were treated for schistosome infection. Mollusciciding and environmental modification were made for eliminating snails at an area of 3,634,580 m2 and 67,105.5 m2 respectively. The length of concrete irrigation and piping system installed was 51.13 and 104.895 km respectively. Methane-generating tanks, water heating instruments using solar energy and other sanitary facilities were established in 1,781 households. After three-year intervention, no infected snails were found and no infected human being, cattle and wild rats were detected. CONCLUSION: Schistosomiasis control was financially supported since the very beginning of the Ertan Dam project, which provided a condition for sustainable development. Continued surveillance of snails and infection sources should be carried out, which will provide scientific basis for schistosomiasis control in the Three Gorges region as well as other new projects of hydropower and water conservancy in endemic area. PMID- 12571970 TI - [The role of Anopheles anthropophagus in malaria transmission in in Xinyang City of Henan Province]. AB - OBJECTIVE: To study the role of Anopheles anthropophagus in malaria transmission and transmission threshold so as to provide basis for vector surveillance and malaria control strategy. METHODS: Parasitological and entomological methods were used in the investigation at 5 villages of Xinyang City, Henan Province. RESULTS: From July to August, 1999, 74 febrile cases (10.9% of the total population) were examined. Among them 50 were infected, the incidence in the population of surveyed spots was 7.4%. Active detection was made in another randomly selected two villages and found that the parasite rate in the inhabitants was 2.0%, and the positive rate of IFA was 8.4%. Only vivax malaria was detected. An. anthropophagus and An. sinensis were collected, with An. anthropophagus as the predominant one in human dwellings. The estimated man-biting rate and the human blood index were 4.9388 and 0.7858 respectively. The vectorial capacity of An. anthropophagus was 5.5296. The critical man-biting rate of An. anthropophagus was 0.2407 as calculated by the formula (ma = -rlnP/abPh) according to Macdonald's model. The local man-biting rate was 20 times higher than that of the critical man-biting rate. CONCLUSION: The results demonstrated that An. anthropophagus is the principal vector in malaria transmission in the area. The findings imply that the critical man-biting rate is of practicable importance in vector surveillance. PMID- 12571972 TI - [Experimental study on the pathogenesis of Entamoeba gingivalis]. AB - OBJECTIVE: To study the pathogenesis of Entamoeba gingivalis (E. g.) and its relation to periodontal diseases. METHODS: Rats were treated with immuno inhibitor for one week and the neck of incisor teeth of the rats was bound with steel wire. They were randomly divided into three groups: the first group was infected by E. g. in the periodontal tissue, the second group was infected by symbiotic bacteria (s. b.), and the third group was given physiological saline as control. Observation on the periodontal inflammation was made for each group of rats, and the purulent secretion from periodontal abscess was examined for living pathogens. RESULTS: The incidence of periodontal diseases in rats infected by E. g. was higher than that of symbiotic bacteria group and that of control (P < 0.05), the incidence of periodontal diseases in rats infected by s. b. was higher than that of control group (P < 0.05). Living pathogens were found in the abscess liquid. CONCLUSION: E. g. is an opportunistic pathogen, which, together with synergistic symbiotic bacteria, can cause periodontal diseases in hosts with low immunity. PMID- 12571973 TI - [Effect of experimental infection with Schistosoma japonicum on the pregnancy of mice]. AB - OBJECTIVE: To understand the effect of Schistosoma infection on the gestation in mice. METHODS: Female mice infected with Schistosoma japonicum cercariae, and mated with male mice (uninfected) at 40 d and 100 d post-infection, the changes during pregnant period and the growth of offspring were observed until birth. The serum level of estradiol and progesterone of the infected mice was measured by RIA at oestrus. RESULTS: The level of estradiol and progesterone, and the pregnant rate were much lower in schistosome infected group than that of the control. The rate of abortion, the mortality of pregnant mice and the death rate due to abortion of infected mice increased significantly. The mortality increased with the time of merging male and female mice in one cage prolonged. The body weight and length of the offspring in both infected and control groups were found no significant difference. CONCLUSION: The results revealed that schistosome infection may suppress estradiol and progesterone secretion, decrease the rate of pregnancy, and that it may also increase the complications and mortality during the gestation periods. PMID- 12571974 TI - [Pathological changes of diffuse Pneumocystis carinii infection in the liver of an AIDS patient]. AB - OBJECTIVE: To examine the pathological changes in the liver of an AIDS patient with complicated infection of Pneumocystis carinii(PC). METHODS: A liver biopsy was made. The tissue was stained with HE, PAS, Giemsa, GMS, and acid-fast staining, and examined under light microscope and transmission electron microscope. RESULTS: Granulomas (acid-fast negative) in the tissue and numerous pathogens (PAS positive) in hepatic sinusoids were detected. Giemsa and GMS staining and electron microscopy all confirmed that the pathogen was Pneumocystis carinii. CONCLUSION: The pathological findings revealed a diffuse extrapulmonary infection of Pneumocystis carinii in the patient of AIDS. PMID- 12571975 TI - [Investigation on the prevalence of human Demodex among 2,248 medical students in inner Mongolia]. AB - OBJECTIVE: To understand the difference of Demodex prevalence among medical students living and studying together with different classes, sexes and nationalities. METHODS: Demodex folliculorum and Demodex brevis were detected by using cellophane tape on the nasolabial grooves. RESULTS: The overall prevalence was 51.5% in 2,248 medical students. Prevalence in different classes and sexes was: freshman 42.6%, sophomore 49.6% and third year students 66.8%; 55.7% in males and 48.7% in females. Prevalence in students with different nationalities was: Mongolian 49.8%, Han 52.8% and other minorities 64.3%. CONCLUSION: The Demodex prevalence in students of senior classes was higher than those from junior ones (P < 0.01), higher in males than in females (P < 0.01). Prevalence in Han students was slightly higher than in Mongolian with no statistical difference. PMID- 12571976 TI - [Plasmodium falciparum erythocyte membrane protein 1 (PfEMP1) and gene family var]. PMID- 12571977 TI - [CD4+ T cells and protective immunity against schistosome infection]. PMID- 12571978 TI - [Investigation on acute schistosomiasis in Tongling County in 1990-1999]. PMID- 12571979 TI - [Advance in study of immune mechanism to limit the development of Plasmodium in anophelines]. PMID- 12571980 TI - [Diagnosis of schistosomiasis with gastroscope in 3 patients]. PMID- 12571981 TI - [Comparison of three approaches to prevent falciparum malaria in Chinese who worked in an African country with high endemicity of malaria]. PMID- 12571982 TI - [Thirty-three cases of brain cysticercosis with large cysts]. PMID- 12571983 TI - [Mass chemotherapy for intestinal helminth infections in primary and secondary school students in Maoming city]. PMID- 12571984 TI - [Effect of immunostimulatory DNA sequence on the production of Th1 and Th2 cytokines induced by Dermatophagoides farinae allergen in vitro]. AB - OBJECTIVE: To investigate the immunoregulatory effect of immunostimulatory DNA sequence (ISS) on the production of Th1 and Th2 cytokines induced by mite allergen in PBMC of the patients with mite allergic asthma in vitro. METHODS: PBMC from the patients with allergic asthma and normal controls were isolated and cultured in vitro stimulated by ISS and Dermatophagoides farinae allergen (Df). IL-12, IFN-gamma and IL-5 in the cell supernatants were detected by ELISA. Df specific IgE in sera of patients were assayed by fluorescent enzyme immunoassay. RESULTS: PBMC from both the patients and normal controls stimulated by ISS plus Df produced a significant increase in the level of both IFN-gamma and IL-12 compared with non-ISS and Df stimulations, whereas IL-5 was decreased. Moreover, the levels of IFN-gamma, IL-12 produced were significantly higher in normal controls than in the patients, on the contrary, IL-5 was down regulated. It was also shown that the level of IL-12 produced by PBMC of the patients with ISS plus Df stimulation correlated positively with that of IFN-gamma. CONCLUSION: ISS not only promotes the expressions of Th1 cytokines but also downregulates the production of Th2 cytokines induced by Df in both allergic and non-allergic individuals, indicating its potential application in the immunotherapy of mite allergy. PMID- 12571985 TI - [Screening and sequence analysis of the adult stage-specific gene of Trichinella spiralis]. AB - OBJECTIVE: To obtain the sequence of the adult stage-specific gene of Trichinella spiralis. METHODS: An adult T. spiralis cDNA library was screened using an adult T. spiralis stage-specific probe. The gene sequence was analyzed by DNAstar software and GenBank database. RESULTS: A cDNA clone of 1,629 bp was obtained by screening the library with an adult stage-specific probe. The clone contained one open reading frame composed of 1,464 bp encoding 487 amino acids. GenBank searching revealed that no identical cDNA was found. Two zinc fingers located at the positions within amino acids 207-270. Some possible antigen epitopes were predicted within the amino acids 52-64, 108-116, 137-163 and 226-260. CONCLUSION: A novel stage-specific gene of adult T. spiralis encoding the protein containing certain predicted antigen epitopes was obtained. PMID- 12571986 TI - [Diagnosis and treatment of a case with cerebral falciparum malaria]. PMID- 12571987 TI - [Exogenous nitric oxide induces apoptosis in tachyzoites of Toxoplasma gondii]. AB - OBJECTIVE: To explore whether NO is able to induce apoptosis in Toxoplasma gondii tachyzoites. METHODS: Apoptosis induced by NO in T. gondii tachyzoites was investigated by TUNEL (terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling) method, electron microscopy and agarose gel electrophoresis. RESULTS: NO donor, sodium nitroprusside (SNP), was found to induce apoptosis in Toxoplasma gondii tachyzoites in a time- and dose-dependent manner by TUNEL detection. N-acetylcysteine, a NO scavenger, could inhibit SNP-induced apoptosis in the tachyzoites while potassium ferricyanide could not induce apoptosis in the tachyzoite. Electron macroscopy showed that SNP-treated tachyzoites possessed typical morphological features of apoptosis, including chromatin condensation below the nuclear membrane, nuclear pyknosis, and formation of apoptotic body. Agarose gel electrophoresis revealed that SNP-treated tachyzoite DNA fragment exhibited characteristic "DNA ladder" after 15 to 20 h. CONCLUSION: SNP, NO donor, might induce apoptosis in T. gondii tachyzoites in terms of characteristic morphological and biochemical features. PMID- 12571989 TI - [Relationship between the distribution of Anopheles virus and Anopheles minimus and malaria in the reclamation area of Hainan Province]. PMID- 12571988 TI - [Studies on synthesis and degradation of collagen at transcription level in liver fibrosis of rabbits with schistosomiasis japonica]. AB - OBJECTIVE: To study the synthesis and degradation of collagen at the transcription level during liver fibrosis in rabbits with schistosomiasis japonica. METHODS: New Zealand rabbits infected with Schistosoma japonicum cercariae were served as animal models of liver fibrosis. The liver specimens were collected through operations at 4, 6, 8, 10, 12, 16, 20, 24 and 28 weeks after infection. Type I collagen, type III collagen, type IV collagen, MMP-1 and MMP-9 mRNA levels of liver tissue were detected by RT-PCR plus dot blotting, and the size of egg granulomas and the degree of liver fibrosis were measured by histopathological examinations. RESULTS: Type I collagen, type III collagen, type IV collagen, MMP-1 and MMP-9 mRNA levels increased simultaneously in the early stage after the infection, mostly reaching their peaks at 10 weeks after infection. Compared with normal controls, type I collagen, type III collagen, type IV collagen, MMP-1 and MMP-9 mRNA levels increased by 12.0-, 11.0-, 6.6-, 10.0- and 11.0-fold, respectively, coinciding with the changes of egg granulomas. Thereafter, both collagen and collagenase mRNA levels decreased. Types I, III and IV collagen mRNA levels declined to 2-fold to 3-fold compared with normal controls (P < 0.05), while MMP-1 and MMP-9 mRNA levels declined to normal level (P > 0.05) at 28 weeks. This study showed that the synthesis and degradation of collagen remained dynamic balance in the early stage of schistosomiasis, while in the later stage the metabolism of collagen synthesis was higher than that of collagen degradation. CONCLUSION: It was confirmed at the transcription level that when the metabolism of collagen synthesis was higher than that of collagen degradation in rabbits with schistosomiasis japonica, liver fibrosis might be produced. PMID- 12571990 TI - [Expression and immunocompetence characterization of Plasmodium falciparum lactate dehydrogenase]. AB - OBJECTIVE: To express lactate dehydrogenase (LDH) gene of Plasmodium falciparum FCC1/HN in the E. coli TG1 and analyse its immunocompetence. METHODS: The LDH gene of the P. falciparum was specifically amplified by polymerase chain reaction, and the recovered gene fragment was cloned into pGEX-4T-1 vector for expression of fusion protein with glutathione S-transferase(GST). The recombinant plasmid was transformed into the E. coli TG1. Four mice (Kunming strain) were immunized with purified expressed protein(antigen) and the polyclonal antibodies were collected. The immunocompetence of recombinant protein was analysed by ELISA and Western-blot. RESULTS: The LDH gene of P. falciparum was successfully expressed in the E. coli TG1. The expressed protein exhibited a specific reaction with immune sera obtained from rabbits immunized with P. falciparum. The specific humoral responses were induced in mice and the titer of the specific antibody was 1:16 by two-dimensional diffusion assay. CONCLUSION: The LDH gene of P. falciparum has been successfully expressed in the E. coli TG1 and the expressed protein has high antigencity. PMID- 12571991 TI - [Studies on the activity and immunohistochemistry of heme oxygenase in Schistosoma japonicum]. AB - OBJECTIVE: To explore the activity of heme oxygenase and immunolocate the enzyme in the adult worms of Schistosoma japonicum. METHODS: Microsomal protein was isolated from the homogenate of adult S. japonicum, heme degradation and effect of different pH conditions and buffers on degrading reaction were investigated by incubating microsomal protein with hemin. The slices of whole worm and cells of S. japonicum were prepared, distribution of HO in schistosome was studied by immunofluorescent and alkaline phosphatase(AP)-immunocytochemical assays. RESULTS: Microsomal protein of adult worms can degrade the heme in vitro, the activity being 56.7 nmol bilirubin/(mg.min). The optimal pH was 8.7. Immunofluorescent and AP-immunocytochemical assays revealed that the HO distributed dispersively in the worm, and located in cytoplasm. CONCLUSION: The presence of HO was firstly proved in S. japonicum. PMID- 12571992 TI - [Gene point mutation in the dihydrofolate reductase-thymidylate synthase gene of Plasmodium falciparum]. AB - OBJECTIVE: To investigate the gene point mutation in the dihydrofolate reductase thymidylate synthase(dhfr) gene of Plasmodium falciparum isolate from Yunnan Province strongly associated with pyrimethamine and cycloguanil resistance. METHODS: Nested PCR and restriction endonuclease digestion were applied to detect the gene mutation using dried blood filter paper collected from the fields in Yunnan Province. RESULTS: Different mutations were found in 4 amino acids at positions 16, 51, 108 and 164 of dhfr gene, particularly, Asn-108 and Ile-51, the mutation frequency being 94.1% and 90.1%, respectively. The frequency of the wild type genotype (3D7-type) Ser-108 appeared lower (9.1%), while the frequency of the Ala-16 was high(61.8%); the mutation-type was very high, the ratio of HB3 type, 7G8-type/FCR3-type and Cambodian-type was 1:21:7.5. CONCLUSION: The investigation first demonstrated that Plasmodium falciparum Yunnan isolate dihydrofolate reductase-thymidylate synthase gene (dhfr) at positions 16, 51, 108 and 164 exhibited different degrees of point mutation. The frequency of mutation of the 7D8-type involved in pyrimethamine resistance was higher, while that of the FCR3-type involved in cycloquanil resistance was lower. PMID- 12571993 TI - [Cloning and sequencing of cytochrome c oxidase II(COII) gene of three species of mosquitoes]. AB - OBJECTIVE: To determine and compare the complete sequence of COII gene from Anopheles sinensis, Aedes albopictus and Culex quinquefasciatus and to identify the molecular homology among these mosquitoes. METHODS: The recombinant plasmids including COII gene from the three species of mosquitoes were obtained by the T-A cloning technique, and the COII genes were sequenced and analyzed. RESULTS: The homology of the COII gene nucleotide sequence and the deduced amino acid sequence of COII among the three species of mosquitoes varied from 84.1% to 87.9% and from 85.1% to 89.5%, respectively. The C + G content of COII gene was 23.2%-24.9%, the frequency of transversion higher than that of transition. CONCLUSION: Aedes albopictus and Culex quinquefasciatus are closely related species at their COII molecular level according to COII gene. PMID- 12571994 TI - [A case with myiasis]. PMID- 12571995 TI - Investigation of risk factors for development of human hydatidosis among households raising livestock in Tibetan areas of western Sichuan province. AB - OBJECTIVE: To identify factors influencing the development of hydatidosis in Tibetan areas of western Sichuan. METHODS: A questionnaire investigation was carried out to collect data on factors related to hydatidosis among households raising livestock. RESULTS: Analyses of data revealed that dogs rather than foxes were the most important source of AE transmission. People below 19 years old tended to suffering from CE rather than AE and people 19-38 years old, especially the females, were under an increasing risk of suffering from AE (female vs. male, OR = 2.438, CI = 1.317-4.514, P < 0.05). Increased risks of both AE and CE prevalence associated with nomadic life, aging, playing with dogs, not protecting food from flies, and raising yaks or sheep. CONCLUSION: In addition to age, sex, environmental conditions and hygienic behaviors, raising yaks or sheep and playing with dogs also increase the risk of suffering from hydatidosis as well in this area. PMID- 12571996 TI - [Application of dot immunogold filtration assay for detecting serum antibodies in clonorchiasis patients]. AB - OBJECTIVE: To establish a new, fast, sensitive and specific assay for detecting antibodies in clonorchiasis. METHODS: Using Clonorchis sinensis adult worm antigen and SPA labelled with colloidal gold as color developing agent, a dot immunogold filtration assay (DIGFA) for detecting clonorchiasis antibodies was tested. Dot-ELISA was used as the parallel control. RESULTS: The positive rate of DIGFA and dot-ELISA in testing the sera from 119 proved cases were 96.4% (115/119) and 92.4% (110/119), respectively, the difference between the two assays being in significant (P > 0.05). The negative rate of DIGFA in healthy people was 100% (40/40). The cross reaction rate in 20 cysticercosis cases and 25 schistosomiasis cases were 5% (1/20) and 4% (1/25), respectively. Both coincidence rates comparing DIGFA with dot-ELISA were 90.9% (50/55). CONCLUSION: DIGFA is as sensitive and specific as the dot-ELISA, and has the advantages of simplicity and without specific equipment. PMID- 12571998 TI - [Studies on detecting Brugia malayi larva in mosquitoes by polymerase chain reaction]. AB - OBJECTIVE: To establish a specific, sensitive and simple assay for the detection of Brugia malayi larva in Anopheles sinensis. METHODS: Using a new DNA purification technique (Microcon 100) and two pairs of oligonucleotide primers (p1, p2 and p3, p4) suitable for detecting B. malayi in seven areas in our country, the mosquito vectors infected by B. malayi were detected by polymerase chain reaction(PCR). RESULTS: This PCR method could amplify separately a 322 basepair(bp) and a 155 bp DNA fragment and detect as few as 1/64 of one L1 in 1 mosquito, the detectable limit was nearly 4 pg DNA of filarial larvae, and it could also detect 1 infected mosquito with one L3 of B. malayi in pools of up to 200 mosquitoes. In contrast, no such specific 322 bp or 155 bp DNA band was detected in Dilofilaria immitis and normal mosquito. CONCLUSION: This PCR technique established for supervision of mosquito vector in B. malayi endemic areas is specific, sensitive, and simple. PMID- 12571997 TI - [Random amplified polymorphic DNA analysis of Cryptosporidium species and strains]. AB - OBJECTIVE: To analyse the genetic heterogeneity of 8 isolates of Cryptosporidium belonging to three species (Cryptosporiduim muris, Cryptosporiduim parvum and Cryptosporidium meleagridis) using RAPD technique. METHODS: DNA extracts of the 8 isolates were amplified by PCR using 26 out of 200 primers. The amplified products were separated by agar electrophoresis followed by staining with ethidium bromide. RESULTS: RAPD analysis revealed; 1. Obvious differences among species were found, the average genetic distance being 7.037; 2. Certain differences among different isolates of the same species were found, the average genetic distance being 5.162. CONCLUSION: RAPD could differentiate the common and respective gene fragments of the 8 isolates of Cryptosporidium. PMID- 12571999 TI - [Studies on the sex difference in proteins between male and female adult worm of Schistosoma japonicum]. AB - OBJECTIVE: To explore the sex difference in proteins between male and female adult worm of Schistosoma japonicum. METHODS: Two-electrophoresis was used to analyse the difference of protein between the male and female adult worm of S. japonicum (Chinese strain). RESULTS: Two-dimensional electrophoresis analysis revealed that at the site of 43 kDa and an isoelectric point (pI) of 5.60-5.90 the male worm exhibited a band carrying a number of spots and dots, being longer and wider than that exhibited by the female worm. The female worm exhibited 7 specific dots. CONCLUSION: The sex differences in proteins between male and female adult worms of S. japonicum are significant. PMID- 12572000 TI - [Pathological changes and host tissue reaction in alveolar echinococcosis]. PMID- 12572001 TI - A brief introduction to the research achievement on the strategy and technical measures for interrupting the transmission of lymphatic filariasis in China. PMID- 12572002 TI - [Survey of Cryptosporidium infection in three areas in Gansu Province]. PMID- 12572003 TI - [An exploration on the reform in the teaching of parasitology in medical institutions]. PMID- 12572004 TI - [Clinical analysis of misdiagnosis of 24 cases of cerebral cysticercosis]. PMID- 12572005 TI - [The progress and direction of medical malacological research]. PMID- 12572006 TI - [Recent progress in the research of immunology of Toxoplasma infection]. PMID- 12572007 TI - [Clinical significance of detection of serum tumor necrosis factor-alpha in children with cerebral cysticercosis]. PMID- 12572008 TI - [Microplate test of non-specific esterase detection for insecticide resistance of Culex pipiens pallens]. PMID- 12572009 TI - [Studies on the molluscicidal effect of Dioscorea zingiberensis against Oncomelania hupensis]. PMID- 12572010 TI - [Efficacy of tinidazole-sulfur cream in the treatment of Demodex infection]. PMID- 12572011 TI - [Clinical analysis of 709 cases of cystic echinococcosis in Xizang autonomous region]. PMID- 12572012 TI - [Genotypes of merozoite surface protein 2 of Plasmodium falciparum in different malaria endemic areas]. AB - OBJECTIVE: To determine the genotypes and distribution of MSP2 of Plasmodium falciparum isolates in Yunnan and Hainan provinces, China. METHODS: The central polymorphic region of MSP2 allele was amplified by the nested PCR for genotyping of P. falciparum. RESULTS: The higher degree of polymorphism of MSP2 of P. falciparum was observed in Yunnan and Hainan. Distribution and allele frequencies differed in both provinces, indicating considerable geographical heterogeneity of parasite populations. The mixed infection of different allele-type and multiplicity of infection was more frequent in Hainan than in Yunnan. CONCLUSION: There were obvious differences in the distribution and frequencies of MSP2 alleles between Yunnan and Hainan endemic areas. MSP2 is suitable to be used as a marker gene for the genotyping of P. falciparum infection. PMID- 12572013 TI - [Gene typing of merozoite surface protein 1 of Plasmodium falciparum isolates from Hainan Province]. AB - OBJECTIVE: To identify the genotype of merozoite surface protein 1 (MSP1) of Plasmodium falciparum isolates from Hainan Province. METHODS: Nested PCR was applied to amplify the MSP1 of Blocks 2 and 3 Plasmodium falciparum isolates from Hainan Province. Two allelic family representative gene fragments were sequenced. RESULTS: From 36 out of 39 blood samples from Plasmodium falciparum patients, 44 gene fragments of blocks 2 and 3 of the MSP1 were amplified, of which the MAD20 type allele was dominant(75%). followed by K1-type allele. No RO33-type allele was found. The mixed infection rate of the two different allelic type was 19.4%. Sequence analysis showed that the sequences of MAD20- and K1-type isolates from Hainan Province were highly homologous to that of the MAD20 and K1 allelic prototypes. CONCLUSION: Two principal allelic types of MSP1 gene, MAD20- and K1 type, exist in malaria endemic areas in Hainan Province, the MAD20-type being the dominant. PMID- 12572014 TI - [Comparison of immune responses elicited by recombinant protein and eukaryotic expression plasmid based on histidine rich protein 2 of Plasmodium falciparum]. AB - OBJECTIVE: To identify the immune characteristics of different vaccine prototypes based on HRP2 and to provide experimental evidence for developing P. f. blood stage vaccines. METHODS: BALB/c mice were immunized with recombinant protein TP HRP2 or eukaryotic expression plasmid pcDNA3.1(-)/HRP2. The kinetics and specificities of antibody responses were analyzed. The proliferation tests of spleen cells were done, and P. f. growth inhibition assays were done with immune sera. RESULTS: The mice immunized with TP-HRP2 in Freund's adjuvant produced high level and high-specificity antibody response. The antibodies appeared rapidly and lasted for a longer time. Cellular responses were induced simultaneously, and the immune sera could inhibit the development of parasite in IRBCs. The mice immunized with pcDNA3.1(-)/HRP2 produced middle-level antibody response which had some specificity, however, the induction of antibodies required repeated inoculation and a longer duration. Immune cells were well primed and the memorial immune response was obvious but the immune sera had no effect on the growth of P.f. in vitro. CONCLUSION: Both the recombinant protein and plasmid DNA based on HRP2 have different immune characteristics in mice. HRP2 recombinant protein has the potential in practical application. PMID- 12572015 TI - [Screening the mimic antigen epitopes of triosephosphate isomerase of Schistosoma japonicum Chinese strain (Sjc-Tpi) with random phage peptide library]. AB - OBJECTIVE: To screen the mimic antigen epitopes of the triose phosphate isomerase of Schistosoma japonicum Chinese strain (SjC-TPI) and investigate their immunogenicity. METHODS: The random phage peptide library (PH.D.-12) was screened with the purified antibody(IgG) against SjC-TPI to get the positive phage which contained the mimic antigen epitopes of SjC-TPI, and the immuno-characterization of the mimic antigen epitopes were investigated. RESULTS: Two mimic antigen epitopes (M1, M2) of SjC-TPI were obtained. The immuno-sera of mice (Kunming strain) against the positive phages could recognize both the SjC-TPI and the protein of the positive phages. The DNA sequencing data showed no homology between the sequences of the deduced amino acid of the two mimic antigen peptides and the amino acid of SjC-TPI. CONCLUSION: The two mimic antigen epitopes of SjC TPI obtained are imitative epitopes of the configuration antigen of SjC-TPI. PMID- 12572016 TI - Cloning and expressing of the three repeat fragments of Plasmodium falciparum 11.1 gene. AB - OBJECTIVE: To clone and express the 3R, 6R and 9R repeat fragments of Plasmodium falciparum(Pf11.1) gene. METHODS: Three repeat fragments from the genomic DNA of Plasmodium falciparum 3D7 strain cultivated were amplified by using the designed primers. The PCR products were cloned into the pT7 vector for bi-direction sequencing. The sequencing results were analysised by GENETYX-MAC. And then the amplified fragments were subcloned into pET32a(+) or pET32b(+) in order to express the recombinant proteins under the induction of IPTG in E. coli BL21. RESULTS: 3R,6R and 9R fragments with sizez of 552 bp, 630 bp and 444 bp respectively were successfully amplified by PCR. The sequence analysis showed that there were 4 more 3AA units and one more 6AA unit in Pf11.1 gene of 3D7 strain as compared with Palo Alto strain. The homologies of the nucleotide sequence between the 3R fragment and the 6R fragment of the two strains were 92.8% and 95.1%, respectively. The amplified 9R fragment contains 139AA repeat units. The three recombinant proteins were expressed in BL21 strain with molecular weights of 45, 60 and 42 kDa. CONCLUSION: We got the 3R, 6R and 9R fragments separately by PCR and expressed them in E. coli successfully. The Pf11.1 gene of 3D7 strain is highly homologous to that of the Palo Alto strain. PMID- 12572017 TI - [Analysis of the mitochondria-related protein of Schistosoma japonicum and its antigen epitopes]. AB - OBJECTIVE: To sequence the cloned gene Sj338 and to identify the encoded protein and its antigen epitopes. METHODS: The Sj338 gene fragment obtained from adult S. japonicum cDNA library amplified by PCR method was subcloned into pGEM-T vector for sequencing. The sequence of nucleotides and the characteristics of the encoded protein were analyzed by DNASIS Program and Goldkey DNA and Protein Analytical Program, and then the homology of the amino acid sequence was searched on the BLAST net. RESULTS: The cloned rSj338 gene was demonstrated to be 487 bp containing one 459 bp ORF, encoding a protein consisted of 153 amino acids with a molecular weight of 17.6 kDa. The amino acid sequence of the recombinant protein rSj338 shared 46% identity with that of the corresponding part of human mitochondrial import receptor and 44% identity with that of the Rattus sp. mitochondrial precursor receptor. The possible antigen epitopes were predicted within the peptide fragments of 26-32 aa, 37-46 aa and 147-151 aa. CONCLUSION: The protein encoded by rSj338 gene fragment might be the mitochondria-related protein of Schistosoma japonicum. PMID- 12572018 TI - [Cloning and expression of the gene encoding Schistosoma japonicum tropomyosin]. AB - OBJECTIVE: To clone and express the cDNA encoding Schistosoma japonicum tropomyosin. METHODS: The cDNA was amplified by reverse transcription-polymerase chain reaction (RT-PCR). The PCR products were ligated with pGEM-T vectors and then for transformations. After characterization of white clones by agarose gel electrophoresis, endonucleases digestion and PCR, some recombinant plasmids with inserts were used for sequencing. Then the gene was subcloned into prokaryotic expression vector pQE30 and expression was induced by IPTG. RESULTS: The PCR products was 823 bp judged by agarose gel electrophoresis and sequencing. A cDNA encoding S. japonicum tropomyosin, except for 14 amino acids at the amino terminus and 2 at the carboxyl terminus, has been constructed and cloned successfully. The colony, designated pGSjcTM12, was sequenced and shown to be 91.1% identical at the nuclei acid level and 98.1% identical in deduced amino acid sequence to that of S. mansoni tropomyosin. The gene was subcloned into pQE30 and an expressed protein of about 32 kDa was obtained. CONCLUSION: The cloning and expression of the gene encoding S. japonicum tropomyosin had been successfully made. PMID- 12572019 TI - [Dynamic changes in collagen type I and collagen type III in rabbits infected with Schistosoma japonicum and the effect of gamma-interferon]. AB - OBJECTIVE: To observe the dynamic changes in collagen type I and collagen type III in rabbits with schistosomiasis japonica and the treatment effect of gamma interferon on the degradation of collagens in schistosomal hepatic fibrosis. METHODS: Each rabbit was infected with 80 +/- 1 S. japonicum cercariae. Liver operations were done at different time points after infection and the liver specimens were embedded with paraffin and stained with alpha-SMA, HE and picric acid-Sirius red. The stained slides were observed under polarizing microscope and different collagen areas calculated by computer imagine analysis system. At the 16th week after infection, the infected rabbits received a single dose of praziquantel and gamma-interferon for 8 weeks. RESULTS: The area percent of collagen type I at the 28th week after infection (40.14 +/- 17.00) increased about seven fold compared with the 8th week group (5.73 +/- 3.40). The area percent of collagen type III at the 28th week after infection (6.80 +/- 5.19) increased about six fold compared with the 8th week group (1.15 +/- 1.34). The alpha-SMA positive cells also increased significantly. After gamma-interferon treatment, the area percent of collagen type I and type III decreased significantly, from 18.51 +/- 7.52 and 4.63 +/- 3.64 (before treatment) to 3.09 +/- 1.54 and 0.40 +/- 0.37 (0 and 4 weeks after treatment) (P < 0.01). However, after the withdrawl of gamma-interferon treatment, the collagen degradation was reversible. CONCLUSION: Gamma-interferon is effective in the treatment of hepatic fibrosis in rabbits infected with S. japonicum, the effect being reversible. PMID- 12572020 TI - [Effect of daphnetin on the exo-erythrocytic stage of rodent malaria]. AB - OBJECTIVE: To investigate the activity of daphnetin(DPNT) against the exo erythrocytic stage of rodent malaria. METHODS: Groups of ten male ICR mice were infected by intraperitoneal injection with sporozoites of P. yoelii. Mice were administered daphnetin 0.5 hr postinfection on d0 and once per day for three additional consecutive days(d1-d3) by the i.g. route. The effects of daphnetin at various dosages and those of the combination of daphnetin with primaquine were assessed by the number of mice with negative Giemsa-stained slides from tail blood on the seventh day after infection and by the average number of red blood cells (RBC) infected in 1,000 RBC observed on the eleventh or twelfth day after infection. We also observed the effect of daphnetin on the concentration of Hb in ICR mice. RESULTS: Daphnetin exhibited no detectable antimalarial effect on the exo-erythrocytic stage of P. yoelii, while the antimalarial efficacy of DPNT 50 mg/(kg.d) combined with 5 mg/(kg.d) PQ, was comparable to PQ 10 mg/(kg.d) x 4 d i.g. in mice infected with sporozoites of P. yoelii. The concentration of Hb in ICR mice administered with DPNT 50 mg/(kg.d) x 4 d decreased on the eighth day after administration. CONCLUSION: Daphnetin alone showed no anti-exoerythrocytic activity in vivo. The combination of DPNT 50 mg/(kg.d) with PQ 5 mg/(kg.d) showed promising antimalarial efficacy comparable to that of PQ 10 mg/(kg.d). Administration of DPNT caused anemia in ICR mice. PMID- 12572021 TI - [Effects of anti-idiotypic antibody NP30 on modulation of egg granuloma formation and hepatic fibrosis of schistosomiasis]. AB - OBJECTIVE: To study the effects of the monoclonal anti-idiotypic antibody NP30 active immunization on egg granuloma formation and hepatic fibrosis in Schistosoma japonicum infection. METHODS: ICR mice were actively immunized with NP30 100 micrograms x 3 i.p. every 10 days while the mice in control group were injected with SP2/0 ascites i.p. simultaneously. After cercariae challenging, the mice were killed at the 4th, 8th, 12th, 16th, 20th and 24th week, respectively. Mouse livers were removed and stained histochemically with VG and subjected to immunohistochemical assay of collagen type I, III and fibronectin(FN). The volume of egg granulomas and the content of collagen type I, III and FN were determined quantitatively by NYD-1000 Image Analysis System. RESULTS: The volume of egg granulomas in NP30 immunized group was much smaller than that of control group from the 12th week after cercariae challenge. The cellular components of egg granulomas in NP30 immunized group were significantly different from those of the control group, exhibiting two types of atypical egg granulomas were found. VG stain revealed that the average optical density of collagen in hepatic granulomas of experimental group was lower than that of control group. Immunohistochemical assay revealed that the contents of collagen type I, III and fibronectin in egg granulomas of experimental group were lower than those of control group. CONCLUSION: NP30 vaccination may induce both cellular and humoral protective immunity to modulate egg granulomas and suppress liver fibrosis of schistosomiasis japonica. PMID- 12572022 TI - [Cytotoxic effect of Acanthamoeba trophozoite on HeLa cells]. AB - OBJECTIVE: To investigate the cytotoxic effect(CTE) on human cervix cancer HeLa cells induced by five strains of pathogenic free-living Acanthamoeba. METHODS: The cytotoxic effect of five isolates of Acanthamoeba on HeLa cells was investigated by light microscopy and MTT method. RESULTS: The photomicrographs of HeLa cells showed a sequence of cardinal morphological features of apoptosis when HeLa cells were exposed to Acanthamoeba in a time-dependent manner at a ratio of 1:1 for 12 h. MTT method showed more than 50% of tumor cells underwent cytolysis following exposure to A. lugdunensis trophozoites, and only 18% of cells treated with A. polyphaga underwent CTE. The CTE produced by A. lugdunensis and A. quina trophozoites was more rapid than the others, beginning as early as 6 h after coincubation and resulting in cytolysis by 72 h. CONCLUSION: These five strains of Acanthamoeba exhibit cytotoxic effects of varying degrees on HeLa cells, inducing apoptosis. PMID- 12572023 TI - [Preliminary study on cytochrome C oxidase 1 gene of Oncomelania hupensis from Miao River area in Hubei province]. AB - OBJECTIVE: To study the mitochondrial cytochrome C oxidase 1(CO1) gene of Oncomelania snails from Miao River area in Hubei Province. METHODS: Oncomelania snails were collected from Miao River area, including upstream and downstream. Genomic DNA was extracted from the tissue of the snail. PCR was used to amplify a fragment of the CO1 gene. Sequences of the CO1 fragment were determined directly from the purified PCR products by an automated sequencer. Sequences for each individual were assembled and edited using ESEE 3.0 s. A distance matrix was computed using program DNADISt of PHYLIP(3.57). Unrooted maximum likelihood trees were calculated from program FITCH. RESULTS: The amplified CO1 gene of the snail was a fragment of 638 bp in length. Sequence analysis showed that the accumulated variable sites were significant different between upstream and downstream populations, being 29 and 46, respectively. From the number of variable sites in the gene, snails in this area were roughly separated into two groups. Each of them was a mixture of both upstream and downstream snails. Same haplotypes were confirmed to be present among the collected sites along the river. From the distance matrix of sequence divergence, the population upstream vs downstream differed by 0.0221 +/- 0.0105. CONCLUSION: There were more variation in downstream population than that in upstream. Gene flow was identified in these populations. The phylogenetic trees suggest the existence of two groups, but all of them belong to 0. h. hupensis. PMID- 12572024 TI - [Dynamic changes in amino acid and glucose in culture medium with adult Schistosoma japonicum]. AB - OBJECTIVE: To observe the dynamic changes in the contents of amino acid(AA), glucose(Gluc) and triglyceride(TG) in the culture medium containing adult Schistosoma japonicum. METHODS: The contents of AA, Gluc and TG in the culture medium during the incubation period for d0 to 6 d were detected by amino acid automatic analyzer and automatic biochemical analyzer. RESULTS: The contents of Arg, Thr, Met, and Lys and Gluc were reduced, Asp and Ala increased apparently. CONCLUASION: Increasing the levels of Arg, Thr, Met Lys and Gluc, reducing the levels of Asp and Ala, and changing the culture medium in time might be in favor of the in vitro cultivation of S. japonicum. PMID- 12572025 TI - [Progress in the research on the techniques for surveillance of filariasis vector]. PMID- 12572026 TI - [Studies on the preventive effect of artesunate against Schistosoma japonicum infection]. PMID- 12572027 TI - [Detection of Cysticercus cellulosae using PCR and DIG-labelled DNA probe]. PMID- 12572028 TI - [A modified method for preparation of Trichomonas vaginalis specimen]. PMID- 12572029 TI - [Immunodiagnostic study on protoscolex antigen of Echinococcus multicularis]. PMID- 12572030 TI - [Detection of canine visceral leishmaniasis by rK 39 antigen dipstick method]. PMID- 12572031 TI - [Efficacy of mandelic acid in treating acute toxoplasmosis in mice]. PMID- 12572032 TI - [A case of skin myiasis caused by Gasterophilus nigricornis]. PMID- 12572033 TI - [Malaria survillance in Sance County, Guizhou Province during 1998-9]. PMID- 12572034 TI - [Investigation on genotype constitution of different Plasmodium vivax isolates and its geographical distribution in China]. AB - OBJECTIVE: To investigate the population constitution and geographical distribution of Plasmodium vivax in China using molecular technique. METHODS: Blood-spot filter paper samples with related epidemiological data were collected from vivax malaria patients living in malarious area of 10 provinces (autonomous region) in China. Semi-nested- or nested-allelic-specific PCR genotyping method was used to identify CSP genotypes, families and types of Plasmodium vivax of each isolate from these patients. RESULTS: Of 384 field isolates of Plasmodium vivax, 258 temperate zone family strains were identified, including 14 allelic variant genotypes spreading among 10 sampling provinces; allelic variants sized less than 731 bp were only seen in 5 provinces in southern China; 79 tropical zone family strains including 5 genotypes were also distributed in 5 provinces of southern China south to 25 degrees N. lat; and 14 PV Type-2 strains including 2 genotypes were found in some areas of Hainan and Yunnan Provinces. In addition, 33 isolates from genotype-mixed infections were revealed. CONCLUSION: At present, area north to 25 degrees N. lat. of the country is the sole area prevalent for Plasmodium vivax family strains of temperate zone; there is overlapping distribution of P. v. of temperate zone family and tropical zone family of this parasite in the southern China south to 25 degrees N. lat; where the most complex isolate constitution is in Yunnan and Hainan Provinces, and PV Type-2 strains have been found in some areas of the two provinces. Besides, there were 2 groups of genotype with distinct geographic distribution feature within the temperate zone family. PMID- 12572035 TI - [Anaphylactic shock caused by effusion of hydatid fluid]. PMID- 12572036 TI - [A study on the recombinant 26 kDa glutathione-S-transferase as a vaccine candidate: dynamics of antibodies in immunized buffaloes and protection against Schistosoma japonicum infections]. AB - OBJECTIVE: To observe the dynamics of antibodies and protection against Schistosoma japonicum infections in buffaloes after immunized with recombinant 26 kDa glutathione-S-transferase (reSjc26GST). METHODS: Buffaloes in 2 villages endemic for schistosomiasis japonica were selected as test and control groups, respectively. In test group initially 96 buffaloes were vaccinated with reSjc26GST, and 90 buffaloes in the control group did not experience vaccination. The indicators included levels of antibodies to reSjc26GST in buffaloes before and after infection with S. japonicum and changes in infection rate. RESULTS: Specific antibodies, which showed a trend of trapezoid increase, were induced in buffaloes after immunized with reSjc26GST. Twenty months after immunization, the infection rate of the test group was decreased by 62.2% when compared with that before vaccination, and by 67.7% when compared with that of the control in the corresponding period. CONCLUSION: Specific antibodies and a certain extent of protection were induced in buffaloes after immunized with reSjc26GST, which played an significant role in ameliorating morbidity. PMID- 12572037 TI - [An outbreak of dermatitis induced by gamasides in Mudanjiang City]. PMID- 12572038 TI - [Immunity against Schistosoma japonicum induced by phage display peptides mimicking antigenic epitopes of Trichinella spiralis]. AB - OBJECTIVE: To obtain the short peptides mimicking antigenic epitopes of Trichinella spiralis (T. s.), and explore their cross protective immunity against Schistosoma japonicum (S.j.) in mice. METHODS: IgG antibodies were purified from sera of mice infected with T.s.. The purified IgG was used to immunoscreen a phage random peptide library of 7 amino-acid residues displayed as a fusion to protein of filamentous phage. Positive clones were obtained by affinity selection, the reactivity of each clone binding to specific IgG was detected by ELISA. Kunming mice were immunized subcutaneously three times with mixed phage clones. The mice were sacrificed 45 days after challenge. The worms and the liver eggs were counted. RESULTS: After three rounds of panning, the relevant phages had been enriched approximately 150 times in production as compared to those from the first round. Of 24 phage clones randomly selected from the third round biopanning, 21 clones were shown to actually bind to the specific IgG. As compared with the control group, the worm and the liver egg reduction rates in vaccination group were 42.8% and 66.3% (P < 0.001), respectively. CONCLUSION: The above results demonstrate that antigenic epitopes of T. s. can be prepared by immunoscreening phage random peptide library and a significant protective immunity against S. j. can be induced by these epitopes in mice. PMID- 12572039 TI - [Two cases of paragonimiasis]. PMID- 12572040 TI - [Clonning and identification of the gene encoding specific IgE antibody-related antigen of Schistosoma japonicum]. AB - OBJECTIVE: To obtain the gene encoding specific IgE-related antigen of Schistosoma japonicum from the cDNA library of adult Schistosoma japonicum. METHODS: The pooled sera from 15 individuals with high levels of specific IgE antibody against SWAP were absorbed with Protein-G and used for screening of IgE related antigen from the adult worm cDNA library of Schistosoma japonicum. The inserted cDNA was amplified by PCR and sequenced. According to the first reading frame of the sequence, a pair of new probes, in which EcoR I and Not I sites were incorporated respectively, were designed and used to amplify the target gene. Then, the gene was cloned into vector pGEM-T and subcloned into expression vector pGEX-6p-1. The fusion protein was expressed, analysed by SDS-PAGE and identified by Western blotting with the specific IgE antibody, respectively. RESULTS: The inserted cDNA fragment from the positive clone was about 1,200 bp, with the ORF of 507 bp which encoded 169 amino acids. The deduced molecular weight of the recombinant protein was 19.3 kDa. The homology between the target gene (Sj43B) and other known DNA sequences was less than 40%. The fusion protein expressed by the recombinant vector pGEX-6p-1/Sj43B could be recognized by schistosome specific IgE antibody. CONCLUSION: Sj43B may encode the specific IgE-related antigen of Schistosoma japonicum. The successful construction of recombinant plasmid pGEX-6p-1/Sj43B lay the groundwork for further studies on immunological characteristics and protection immunity of the recombinant protein. PMID- 12572042 TI - [A case of malaria nephritis]. PMID- 12572041 TI - Studies on antagonism between Exorchis ovariolobularis and Schistosoma japonicum in Oncomelania hupensis. AB - OBJECTIVE: To examine the antagonistic interactions between Schistosoma japonicum and Exorchis ovariolobularis in the snail host, Oncomelania hupensis. METHODS: The infection rate and average number of cercariae of S. japonicum were examined in O. hupensis which had been superinfected with S. japonicum and E. ovariolobularis. The experimental snails were histologically observed by paraffin section. RESULTS: When snails were fed with eggs of E. ovariolobularis 37 days later than S. japonicum infection, the infection rate of S. japonicum (52.9%) was significantly lower than that of the single S. japonicum infected control (75.9%). When E. ovariolobularis infected snails were exposed to S. japonicum miracidia at different intervals, the infection rates of S. japonicum were 64%, 66.7%, 65.2%, 56.4% and 57.1%, respectively. On the other hand, the infection rate of S. japonicum in the single infected control was 90.5%. There existed significant differences. Furthermore, the average number of S. japonicum cercariae shed from superinfected snails were significantly less than that of single S. japonicum infected control. The histological observation showed that the larvae of S. japonicum in superinfected snails were sluggish in development, irregular, shrunken, deformed, relatively small in number as compared with the control. The sporocysts contained few germ balls or immature cercariae and most of the digestive glands of superinfected snails were atrophied. CONCLUSION: There is antagonism between E. ovariolobularis and S. japonicum in O. hupensis, and E. ovariolobularis is dominant in this situation. PMID- 12572043 TI - [The construction of recombinant plasmids of Toxoplasma gondii P30 and the immunoprotective activity of the P30 constructs]. AB - OBJECTIVE: To identify the potential DNA vaccine candidate which can induce the protective immune response to Toxoplasma gondii by inoculating mice with plasmid DNAs encoding three different forms of P30 antigen (membranous secretory, and intracellular). METHODS: Three forms of recombinant plasmid: pcDNA3-P30Mb(contain the whole P30 gene sequence, including the gene encoding signal peptide and hydrophobic tail), pcDNA3-P30Se(contain the whole P30 gene sequence, without the gene encoding hydrophobic tail) and pcDNA3-P30In(contain the whole P30 gene sequence, without the gene encoding signal peptide) were constructed by PCR and subcloning technique. The mice were immunized with different forms of recombinant plasmids and IgG antibodies in the mice were detected by ELISA and Western blotting. RESULTS: Three forms of expression recombinant plasmid of Toxoplasma gondii P30 gene were successfully constructed. The P30 inserts were identified by restrictive enzyme digestion and sequencing. ELISA and Western blotting analysis demonstrated that specific IgG antibody could be induced in three immunized groups, but there was some difference in appearence time and intensity of IgG. CONCLUSION: Genetically immunization of mice with the recombinant plasmids could elicit specific IgG antibodies. In respect to IgG response, the immune efficiency of the three forms of recombinant plasmids was different at the beginning (2 wk), but 4 wk later approximately same. PMID- 12572044 TI - [Studies on ultrastructure and histochemistry of Trichomonas vaginalis adhering to vaginal mucosa of rats]. AB - OBJECTIVE: To study interaction between Trichomonas vaginalis and epithelium of genital tract of host as well as the pathogenesis of T. vaginalis. METHODS: Immunohistochemical technique was used to observe the adhering process of T. vaginalis to vaginal mucosa in rats by transmission and scanning microscopy. RESULTS: T. vaginalis were shown to be PAS positive and clusters of T. vaginalis were found to adhere to columnar epithelium rich in mucopolysaccharide on the surface of vaginal mucosa as viewed in sections of the middle and upper parts of the orgen. T. vaginalis was positive for cathepsin. And the membrane of epithelial cells was often damaged by the released hydrolase. The parasite was also positive for actin; the microfilament bundles were arranged in reticular form in ameboid T. vaginalis. The latter would penetrate between epithelial cells, and its filiform pseudopodia would invade the interspace of microvilli of the epithelium, to encircle and gradually phagocytize microvilli. Digitiform pseudopodia would insert between epithelial cells and encircle part of them. A few T. vaginalis were found to adhere to the keratinized epithelium between mucosal folds as shown in the sections of the lower part of vagina. CONCLUSION: T. vaginalis is inclined to parasitise vaginal fornix because the superficial epithelial cells there are rich in mucinogen granules and abundant microvilli exist. After adhering, T. vaginalis releases hydrolase to digest and phagocytize epithelium which may directly damage the epithelium of the genital canal. Moreover, T. vaginalis would take in mucopolysaccharide to affect the normal clearance process of vagina, resulting in inflammation of parasitized tissue. The cytoskeleton, cell coat, polymorphism of pseudopodia and lysosome of T. vaginalis play an important role in the courses of movement, adhesion, encirclement, phagocytosis and digestion. PMID- 12572045 TI - [Analysis of Leishmania species and strains from China by RAPD technique]. AB - OBJECTIVE: To analyze genetic relationship of Leishmania species and strains from China by RAPD technique. METHODS: DNAs from Leishmania strains, including L. donovani (L. d.) isolates from patients, dogs and sandflies of three different foci in China and international reference strains, were amplified by seven random primers. The DNA polymorphic bands detected were analyzed by clustering analysis with SPSS software. RESULTS: 1. L. d. isolates from hill and plain foci in China were divided into two groups. The genetic distance of L. d. isolates is distant between them. 2. L. d. XJ771, L. d. XJ901, L. d. XJ801 from desert, vicinity of desert, and plain regions in Xinjiang were in the same group. It indicated that the genetic distance among L. d. isolates from the three regions is close. 3. L. d. isolated from VL patients and dogs in hill foci could not be discriminated distintly, showing high homology between them. 4. L. d. DD8 from India, the reference strain of plain type, was clustered with L. d. isolates from plain foci in China. It provided scientific basis for the viewpoint "Kala Azar from east area of China is similar to that from India". 5. L. infantum and L. d. isolates from hill foci in China were clustered into different groups. 6. The genetic distance is close between L. d. isolates from plain foci in China and L. d. Jed; 7. L. infantum and L. tropica showed the closest genetic distance. CONCLUSION: Differences at genetic level exist in Leishmania isolates from different foci in China. PMID- 12572046 TI - [On cleanliness of hands in diminution of Ascaris lumbricoides infection in children]. AB - OBJECTIVE: To study the relationship between cleanliness of children's hands and diminution of Ascaris lumbricoides infection. METHODS: Before the study all persons positive for ascaris eggs in the preliminary survey were treated with albendazole. Hand-washing habit before meal and after defecation was kept in children of experimental group, but not in the control group. Kato thick smear stool examination was done once every two months for one year to compare the new infection rates in children without ascaris infection in the two groups, and the reinfection rates in the cured negative cases were also compared between them in half a month after chemotherapy. RESULTS: All the new infection rates as well as reinfection rates of each reexamination in the experimental group were significantly lower than that of the control group (P < 0.001). Reexamination one year later showed that the ascaris infection rate of the experimental group was 35.2%, reducing by 48.5% as compared with 68.3% before the operation of the project; while ascaris infection rate of the control group was 73.7%, increasing by 78.0% as compared with 41.4% before the operation of the project. CONCLUSION: Washing hands with toilet soap to keep hands clean can significantly reduce ascaris infection rate. PMID- 12572047 TI - [Immunostimulatory activity elicited by CpG sequences in Cysticercus cellulosae paramyosin cDNA]. AB - OBJECTIVE: To analyse the immunostimulatory activity of CpG sequences in cysticercus cellulosae paramyosin (also named Antigen B, AgB) cDNA. METHODS: C57BL/6 mice were immunized with pcDNA3-AgB plasmid, pcDNA3-AgB' (CpG sequences were mutated), pcDNA3 or AgB protein and two weeks later, immune response was assayed by ELISA. RESULTS: IgG and IgG2a were detectable at week 2 after immunization and continually increased until week 4. The antibody levels elicited by pcDNA3-AgB were significantly higher(P < 0.05) than those elicited by others. CONCLUSION: After pcDNA3-AgB plasmid inoculation, the immune response of mouse was elicited not only by the AgB protein but also by the CpG immunostimulatory sequences in the AgB cDNA. PMID- 12572048 TI - [An approach to the morphology of buccal armature of Sergentomyia nankingensis]. AB - OBJECTIVE: To describe the morphological variation of pointed teeth of buccal armature of Sergentomyia nankingensis in a supplementary study. METHODS: The morphology of Sergentomyia nankingensis was observed and the specimens included S. nankingensis captured from the field as well as the female ones bred in the laboratory and their offspring of first generation. RESULTS: The buccal armature of females and males, consists of about 10-19 and 8-15 pointed teeth and the pigmented plate is polymorphous. In the nature, the number of pointed teeth of buccal armature of Sergentomyia is unstable, varying in more than 10 teeth in the same species sometimes. CONCLUSION: Identification of species needs large amount of specimens for comparative observation. Sergentomyia sandfly should be raised separately and individually for morphological observation of its newly emerged offspring, especially when the sandfly specimens and data collected in the field were not sufficient. PMID- 12572049 TI - [Detection of anti-Spirometra erinaceieuropaei antibody using ELISA]. AB - OBJECTIVE: To study specific diagnosis of Spirometra erinaceieuropaei. METHODS: An enzyme-linked immunosorbent assay (ELISA) was studied using highly pure gene engineering antigen expressed by the recombination of the cloned cysteine proteinase gene of Spirametra erinaceieuropaei with expression vector pMAL-c2. Six sera from patient infected with Spirometra erinaceieuropaei were detected using this method. RESULTS AND CONCLUSION: The results showed that the gene engineering antigen reacted strongly with the sera from Spirometra erinaceieuropaei-infected patients, but did not with the sera from Cysticercus cellulosae-infected patients. PMID- 12572050 TI - [A clinical trial of triclabendazole in the treatment of human paragonimiasis skrjabini]. AB - OBJECTIVE: To observe the therapeutic effect of triclabendazole in patients with paragonimiasis skrjabini. METHODS: Four patients were treated with triclabendazole at dosage of 10 mg/kg bid for 3 days and were followed up for 6 10 months. The clinical symptoms, chest X-ray, hemogram and liver function tests were observed before and after treatment. RESULTS: Migratory subcutaneous nodules in three patients all disappeared. Small amount of pleural effusion were absorbed in two patients. Severe headache subsided in a patient with cerebral lesion. MRI showed a bean-sized calcified spot in the right frontal region. Chest X-ray in 2 patients showed that pulmonary lesions were markedly absorbed and disappeared. There was no adverse effect of the drug. CONCLUSION: Triclabendazole has marked therapeutic effect on paragonimiasis skrjabini. It is safe in administration and well tolerated. PMID- 12572051 TI - [A case of American cutaneous leishmaniasis imported from Venezuela]. PMID- 12572052 TI - [A study of artemether combined with primaquine in the treatment of falciparum malaria]. AB - OBJECTIVE: To evaluate the efficacy and side effects of artemether combined with primaquine in the treatment of falciparum malaria. METHODS: Randomization and comparison methods were used in 121 falciparum malaria cases in the Republic of Central Africa. Sixty-one cases were treated with artemether combined with primaquine (Group A used artemether orally, Group B used artemether intramuscularly). And 60 cases received single artemether (Group C used artemether orally, Group D used artemether intramuscularly) were taken as control. RESULTS: In Group A and B the mean fever clearance time were 47.6 +/- 15.7 and 36.9 +/- 10.7 hours, clinical cure rates 84.4% and 100%, relapse rates 6.3% and 3.4%, respectively. In Group C and D the mean fever clearance time were 48.2 +/- 18.4 and 42.2 +/- 9.5 hours, clinical cure rates 90.1% and 96.3%, relapse rates 21.2% and 18.5%, respectively. Side effects in cases of all groups were mild. CONCLUSION: Artemether combined with primaquine and single artemether(via both routes) showed good therapeutic effects in falciparum malaria cases, while artemether combined with primaquine was more effective than single artemether in reducing relapes rate of malaria. PMID- 12572053 TI - [Three lethal cases of Angiostrongylus cantonensis infected children]. AB - OBJECTIVE: To offer approaches to diagnosis and treatment of Angiostrongylus cantonensis infection on the basis of analyzing three lethal cases of the disease. METHODS: Clinical manifestations, pathological changes and species identification were pursued. RESULTS: Angiostrongyliasis is usually neglected in medical departments. The three cases here reported were all misdiagnosed and had not receiced anthelmintic treatment hence fatality ensued. CONCLUSION: Early examination of CSF, bronchial lavage fluid and feces is urged so as to secure accurate diagnosis and conduct efficacious anthelmintic therapy to cure the patients. PMID- 12572054 TI - [Advance in taxonomic researches on Paragonimus sp]. PMID- 12572055 TI - [Hepatic echinococcosis]. PMID- 12572056 TI - [Isolation identification and laboratory evaluation of molluscicidal ingredient 2 hydroxyl-3-butenyl-thiogluco-side from rape seed]. PMID- 12572057 TI - [Relative activity of succinate dehydrogenase in mitochondria of thoracic muscles of Culex tritaeniorhyncus and Culex pipIens pallens]. PMID- 12572058 TI - [Cloning and characterization of three novel genes encoding transmembrane proteins of Schistosoma japonicum]. AB - OBJECTIVE: To clone and analyze novel antigen molecules of Schistosoma japonicum (Sj), and to provide effective vaccine candidate antigens against schistosomiasis japonica. METHODS: Sj adult cDNA library was screened using sera of mice infected with Trichinella spiralis (Ts) and the inserts of positive clones were specifically amplified by PCR. The positive clones were sequenced and the sequence data were analyzed using Nucleotide BLAST software of NCBI and Expert Protein Analysis System of Swiss Institute of Bioinformatics. RESULTS: Nine positive clones were obtained after three rounds of immunoscreening. The size of these inserts ranged from 0.6 kb to 2.1 kb. Among five novel genes, Sj-Ts1, Sj Ts3 and Sj-Ts5 (GenBank accession number: AY005816, AF299080 and AY024352, respectively) encode trans-membrane proteins with 83, 83 and 233 amino acids, respectively. Sj-Ts1 protein predicted contains one possible trans-membrance helix, one N-myristoylation site, two phosphorylation sites for protein kinase C and one for tyrosine kinase, Sj-Ts3 protein contains two possible transmembrance helices and one casein kinase II phosphorylation site, whereas Sj-Ts5 protein has five possible transmembrance helices, one N-glycosylation site, one N myristoylation site, two phosphorylation sites for cAMP- and cGMP-dependent protein kinase and four for protein kinase C and one for casein kinase II. CONCLUSION: Three novel genes encoding three transmembrane proteins might be developed as new vaccine candidates against Sj infection. PMID- 12572059 TI - [A case of roundworm in the splenic artery]. PMID- 12572060 TI - [Enrichment and screening of up-regulated genes of the mosquito Anopheles stephensi in response to malaria parasite]. AB - OBJECTIVE: To isolate and identify genes related to malaria parasite infection in vector mosquito, and to explore the mechanisms. METHODS: Anopheles stephensi infected with Plasmodium yoelii was used as tester (T) group, while uninfected but normal blood fed as driver (D) one. Engorged female mosquitoes of two groups were collected separately at 24 hours after biting. An enriched subtractive cDNA pool was generated through the course of suppression subtractive hybridization (SSH) and selective PCR amplification. The subtracted library was screened by hybridization using T and D cDNA mixture as probes, respectively. The positive clones, which produced stronger signal when probed with T than with D, were sequenced and their sequence homologues in GenBank database were searched with BLAST by internet. RESULTS: The analysis of subtraction efficiency showed that the differentially expressed genes in T comparing to in D were enriched significantly. In dot blot screening, 24 of 58 randomly selected clones (41.4%) were shown up-regulation in malaria infected mosquitoes. The BLAST search of 23 genes revealed that 12 were homologous to functionally known genes, 4 were homologous to functionally unknown entries, and 7 were novel without any relatives. Nine of the 23 genes (39.1%) also hit homologous sequences in the An. gambiae EST database generated from an immune competent cell line treated with lipopolysaccharide (LPS). CONCLUSION: An enriched cDNA pool of the mosquito genes which up-regulated responsively at the early stage of malaria parasite infection was obtained. Expression screening against the pool indicated that various biochemical processes and mechanisms might be involved in the response of mosquito to parasite infection, especially those related with the innate immune system and energy metabolism. PMID- 12572061 TI - [Observation on hemocytes of Culex pipiens quinquefasciatus larvae infected by Lagenidium giganteum]. AB - OBJECTIVE: To explore the possible defensive immunity of Culex pipiens quinquefasciatus larvae after being infected by Lagenidium giganteum. METHODS: To count and analyze morphologically hemocytes of the mosquito larvae with Giemsa's stain and by both phase contrast microscope and ordinary microscope. RESULTS: 1. Following the prolonged duration of infection, the number of hemocytes featured as increase, intense increase and gradual decrease. 2. 48 and 72 h after infection the proportion of plasmatocytes and granulocytes increased significantly, but that of prohemocytes showed a relatively slow increase, while 96 and 120 h after infection the proportion of spherulocytes and oenocytoids increased gradually and that of plasmatocyte and granulocytes showed a slow decrease. 3. After L. giganteum infection, vacuolation and morpholysis were found in plasmatocytes. CONCLUSION: The changes of hemocytes in amount, classification and morphology after infection revealed that Lagenidium giganteum can induce some defensive immune reactions in the larvae of Culex pipiens quinquefasciatus possibly due to a release of antigens or toxic substances by the fungus. PMID- 12572062 TI - [Inhibition of in vitro translation of esterase mRNA of dipterex-resistant mosquito (Culex pipiens pallens) by antisense nucleic acids]. AB - OBJECTIVE: To examine the inhibitory effect of antisense nucleic acid on the in vitro translation of esterase mRNA from dipterex-resistant Culex pipiens pallens. METHODS: 18-mer nucleic acid was synthesized and complementary to the translation initiation site of mRNA of dipterex-resistant mosquitoes. The ODNs were annealed to the corresponding mRNA molecules and they were added to rabbit reticulocyte cell-free system. The translation products were analyzed by SDS-PAGE. After fixing, the gel was exposed to X-ray film by autoradiography for analysis of protein synthesis. RESULTS: Six mumol/L of ODNs elicited a 50% reduction in specific protein expression, and 20 mumol/L of ODNs inhibited the expression of esterase by 80%. The SDS-PAGE showed that the band of reduced amounts of 65 kDa protein for resistant mosquito was almost the same as that for sensitive sample. CONCLUSION: Antisense oligonucleic acids to the esterase mRNA of dipterex resistant mosquito could effectively inhibit its in vitro translation. PMID- 12572063 TI - [Protective immunity induced by 23 kDa membrane protein DNA vaccine of Schistosoma japonicum Chinese strain in mice]. AB - OBJECTIVE: To develop 23 kDa membrane protein DNA vaccine of Schistosoma japonicum Chinese strain and test its protective efficacy in infected C57BL/6 mice. METHODS: The full length cDNA encoding SjC23 amplified from pUC19-SjC23 subcloned into pcDNA3.1. 48 female mice were divided into three groups: A, B and C. Group A (control group) was each immunized i.m. with 100 micrograms of pcDNA3.1; group B (SjC23 group) was each immunized i.m. with 100 micrograms of pcDNA3.1-SjC23; group C (SjC23 + IL-12) was each immunized i.m. with a mixture of 100 micrograms of pcDNA3.1-SjC23, 100 micrograms of pcDNA3.1-p35 and 100 micrograms of pcDNA-p40, followed by two boosts of the same DNA once every two weeks. All the mice were challenged with 45 cercariae at week 8, killed and perfused for worms at week 14. The expression of SjC23 and p35, p40 in muscle tissue was determined by immuno-histochemical method. By the culture of spleen cells, the production of IL-2, IL-4, IL-10 and IFN-gamma after the stimulation of rSjC23-HD was determined two weeks before and after challenge. Anti-SjC23 antibodies were tested by Western blotting. RESULTS: SjC23 and p35, p40 were all expressed on the membrane and in the plasma of muscle cells of the infected mice. Significant increase of IL-2 and IFN-gamma in SjC23 and SjC23 + IL-12 groups was observed before and after challenge. Western blotting showed that after the third immunization (before challenge) 8 out of 10 sera from SjC23 group and 9 out of 10 sera from SjC23 + IL-12 group were positive. The worm reduction rate in SjC23 group and SjC23 + IL-12 group was 26.9% and 35.4%, respectively; the number of eggs in liver tissue was reduced by 22.2% and 28.4%, respectively. CONCLUSION: pcDNA3.1-SjC23 DNA vaccine could induce partial protection against Schistosoma japonicum in C57BL/6 mice. PMID- 12572064 TI - [Environmental investigation in areas where Anopheles anthropophagus distributed in Hubei Province]. AB - OBJECTIVE: To investigate the natural and socio-economical environment in the areas where Anopheles anthropophagus distributed in Hubei Province. METHODS: 5% of the villages in distribution area of An. anthropophagus were randomly selected for the investigation which included vegetation, soil, water, temperature, farming, resting places of the mosquito. RESULTS: The vegetation of An. anthropophagus area consisted of forest, bush, sod and crops. The area was densely covered with river, ditches and ponds, the water pH being 6.1-7.7. The soil texture was either yellow, yellow brown, grey or black, contained 2.72% organic material. There were one or two seasons of rice plantation in the area and the amount of insecticide used was 0.828 kg/mu in the rice field. The annual average temperature was 16.9 degrees C and the humidity was 76.9%. The breed period of An. anthropophagus was from June to September. There are two peaks of mosquito population in area with two seasons of rice plantation, and one peak in one rice season. The composition of An. anthropophagus resting in households, cowsheds and pigpens was 80.9%, 12.2% and 6.0%, respectively. CONCLUSION: The distribution of An. anthropophagus was in point, flaky or belt form in low hill, hillock, shallow hilly plain. These areas were full of vegetation and source of water, the principal crop was single cropping of rice, and soil texture is yellow or yellow brown. The major resting place of the mosquito was human dwellings. PMID- 12572065 TI - [A case with Pthirus pubis parasitizing the eyelash]. PMID- 12572066 TI - [The effect of nitric oxide donor on the DNA content in Toxoplasma gondii tachyzoite]. AB - OBJECTIVE: To determine the role of sodium nitroprusside (SNP) in regulating DNA synthesis of Toxoplasma gondii tachyzoites. METHODS: Hypodiploid peak of tachyzoite DNA induced by SNP was assessed according to DNA fragmentation. The effect of SNP on appearance of hypodiploid peak and the effect of Ca2+ on the growth of tachyzoites were evaluated. The intracellular Ca2+ chelator (BAPTA/AM), antagonist of Ca2+ channel (verapamil) and the extracellular Ca2+ chelator (EGTA) were used. The change of DNA content was measured by flow cytometry. RESULTS: SNP inhibited DNA synthesis of tachyzoites in a dose- and time-dependent pattern. The antiproliferative effect of SNP on tachyzoites was inhibited by verapamil, EGTA and BAPTA/AM. The inhibition of the growth of tachyzoites by SNP was associated with increased subploid peak through a Ca(2+)-dependent mechanism. CONCLUSION: SNP induced a hypodiploid peak in tachyzoites by altering the Ca2+ concentration in the plasma of tachyzoite, resulting in damages of the parasite. PMID- 12572067 TI - [Climbing-upward inhibition of Oncomelania hupensis by niclosamide combination]. AB - OBJECTIVE: To develop a combination of molluscicide/insecticide with a good molluscicidal effect, especially an inhibitory effect on climbing-upward of Oncomelania hupensis. METHODS: Experiments on molluscicidal and snail-climbing inhibition effect of the combination of niclosamide and shachongding [(dimethylamine) trithacyclohexane hydrochloride] were conducted by immersion and spraying in laboratory. RESULTS: The combination of niclosamide (0.2 mg/L) and shachongding (0.1 mg/L) showed an inhibition of 92.67% of snails to climb up. There was no significant difference in molluscicidal effect between the combination (LC90 0.198 mg/L) and niclosamide alone (LC90 0.207 mg/L). CONCLUSION: The combination showed an effective inhibition on the climbing-upward of snails in water so as to improve the molluscicidal effect and reduce the cost of mollusciciding. PMID- 12572068 TI - [Effect of acryl thiourea on liver pathologic changes in mice infected with Schistosoma japonicum]. AB - OBJECTIVE: To observe the effect of acryl thiourea, an inhibitor of phenol oxidase, on pathological changes in the liver of mice infected with Schistosoma japonicum. METHODS: From day 22 to day 42 postinfection with cercariae, the mice of the acryl thiourea group were each injected i.p. with acryl thiourea at a dose of 300 mg/kg every other day. The mice were killed on the 42nd day postinfection to observe the pathological changes in the liver. RESULTS: Compared to the infected control group, the liver tissue of the acryl thiourea group showed scattered foci of inflammatory cell infiltration, the mean diameter and area of the foci were significantly reduced (P < 0.01), and there were no eggs in the center of the foci except for some granules. CONCLUSION: After i.p. injections of acryl thiourea, no typical egg granuloma was found in the liver of infected mice. This was possibly due to the inhibition of schistosome phenol oxidase activity and so the female adult schistosomes could not produce normal eggs. PMID- 12572069 TI - [A case of Clonorchis sinensis infection complicated with gallstone]. PMID- 12572071 TI - [Survey of parasitic diseases in Heilongjiang Province]. PMID- 12572070 TI - [Application of dot immunogold filtration assay in antibody detection for cysticercosis]. AB - OBJECTIVE: To detect serum antibody of cysticercosis by dot immunogold filtration assay for establishing a diagnostic kit for cysticercosis patients. METHODS: Cyst fluid of cysticercus of Taenia solium after dialysis was used as diagnostic antigen in dot immunogold filtration assay and enzyme-linked immunosorbent assay to detect the serum antibody in patients with cysticercosis. Samples to be detected included 71 sera from patients with cysticercosis, 90 sera from healthy people, 20 sera from patients with other parasitic infections or brain tumor. RESULTS: The sensitivity and specificity of dot immunogold filtration assay were 90.1%(64/71) and 95.6%(86/90), respectively. No positive reaction was recorded in cases with other diseases except one serum from a patient with brain tumor. The coincidence rate between dot immunogold filtration assay and enzyme-linked immunosorbent assay was 94.4% (152/161). CONCLUSION: Dot immunogold filtration assay showed promising result for the diagnosis of cysticercosis. PMID- 12572073 TI - [Clinical report of 20 children with cerebral cysticercosis]. PMID- 12572072 TI - [Determination of T lymphocytes and trace elements in spleen from rats infected with Toxoplasma gondii]. AB - OBJECTIVE: To determine the level of five trace elements(Fe2+, Cu2+, Zn2+, Ca2+, Mg2+) in the spleen and changes of T lymphocyte and its subtype variations in peripheral blood from the rats infected with Toxoplasma gondii. METHODS: Twenty rats were randomly and equally divided into two groups: control group and experiment group. Each rat in the experiment group received an i.p. injection of 2 ml normal saline containing 1.5 x 10(6) tachyzoites of T. gondii. On the 64th day after injection of T. gondii, the changes in T lymphocytes (TL) and their subgroups, the helper T lymphocytes (Th) and the suppressor T lymphocytes(Ts) in the peripheral blood of the rats with T. gondii were determined by the assay of the lymphocytes labeled with intercellular acid alpha-naphthyl acetate esterase. All the rats were killed and the atomic absorption method were used for detecting the level of trace elements in the spleen tissue. RESULTS: The number of TL and Th in experiment group was significantly lower than that of control (P < 0.01, P < 0.01). The ratio of Th/Ts showed a significant difference between the two groups. The level of Fe2+, Cu2+ in experiment group was significantly reduced (P < 0.01). The amount of Mg2+ in infected rats was higher than that of the control(P < 0.01). No statistical difference in the content of Zn2+, Ca2+ was found between the two groups. CONCLUSION: T. gondii infection might cause the changes in the TL and Th in peripheral blood and the changes in trace elements in spleen of the rats. PMID- 12572074 TI - [Improvement of amplification method for Cryptosporidium parvum oocysts from mice]. AB - OBJECTIVE: To establish a simplified amplification method for obtaining a large number of purified Cryptosporidium parvum oocysts from infected C57BL/6N mice. METHODS: All mice in the experimental groups were immunosuppressed by given different concentrations of dexamethasone phosphate added in drinking water throughout the experiment. The recovery and purity of the oocysts obtained using different purification methods was compared. The infectivity of the oocysts obtained from the same origin but different animals and different purification methods in a bovine fallopian tube epithelial cell culture system was studied. RESULTS: 4.16 x 10(9) oocysts were obtained in 30 mice in the 3rd group with dexamethasone of 20 micrograms/ml in drinking water. No significant difference in the oocyst recovery, purity and infectivity was found between methods using saturated saline floatation and sucrose density gradient centrifugation. The infectivity of the oocysts obtained from the same origin but different animals was similar. CONCLUSION: A simplified amplification method for obtaining a large number of purified Cryptosporidium parvum oocysts from the infected mice was established. PMID- 12572075 TI - [The Plasmodium digestive vacuole: an antimalarial target]. PMID- 12572076 TI - [Dendritic cells and protozoan infection]. PMID- 12572077 TI - [Studies on polymorphism of MSA-1, MSA-2, and Pf60.1 gene of Plasmodium falciparum in various areas of Yunnan Province]. PMID- 12572078 TI - [Survey of Demodex folliculorum infection in school children in Kaifeng City]. PMID- 12572079 TI - [Cloning and sequence analysis of activated protein kinase C receptor gene of Leishmania major]. PMID- 12572080 TI - [Surgical removal of liver hydatid cyst: a report of 52 cases]. PMID- 12572081 TI - [Expression of circumsporozoite protein gene of Plasmodium falciparum FCC1/HN in BCG]. PMID- 12572082 TI - [Cloning and sequence analysis of exp-1 gene of Plasmodium falciparum FCC1/HN]. PMID- 12572083 TI - [Results and analysis of filariasis surveillance after its elimination in Nanning City]. PMID- 12572084 TI - [Survey of advanced schistosomiasis cases in Hongya City]. PMID- 12572085 TI - [Etiology of community-acquired pneumonia among adult patients in Japan]. AB - To clarify the etiology of community-acquired pneumonia (CAP) in Japan, the causative pathogens were prospectively investigated in adult patients admitted to Kurashiki Central Hospital. The microbiological diagnosis was based on the results of quantitative sputum culture, blood culture, and other invasive procedures, including transthoracic needle aspiration or bronchoscopic examination. Five hundred fifty-two episodes of CAP in 540 patients were admitted between July 1994 and June 1999. Causative pathogens were identified in 353 episodes (63.9%). Several characteristics about the etiology of CAP in Japan were recognized: 1) Streptococcus pneumoniae is the most common pathogen followed by Haemophilus influenzae; 2) Mycoplasma pneumonia is dominant among young patients; 3) Chlomydia pneumoniae is one of the significant pathogens in Japan as well as in western countries; 4) Streptococcus milleri group and anaerobes are important pathogens in patients with suppurative pulmonary diseases; 5) The incidence of Legionella pneumonia is far lower than in western countries; 6) The prevalence of tuberculosis in CAP is still high in Japan; etc. Recognition of these results will lead us to treat patients with prompt antimicrobial therapy. PMID- 12572086 TI - [The role of atypical pathogen: Mycoplasma pneumoniae and Chlamydia pneumoniae in the acute respiratory infection in childhood]. AB - The recent microbiological advance has revealed the importance of atypical pathogens such as Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila as common causes of acute bronchitis and community-acquired pneumonia. We found a third of community-acquired pneumonia in childhood were caused by M. pneumoniae and C. pneumoniae like western countries and there were many dual infections than expected. Therefore we have to treat patients with community-acquired pneumonia in always thinking about the role of atypical pathogens. This article summarizes the epidemiology, specific clinical features, diagnosis, and treatment of these important organisms in the pediatric populations. PMID- 12572087 TI - [In vitro and in vivo antibacterial activities of azithromycin]. AB - In Vitro and in vivo antibacterial activities of azithromycin were compared with those of macrolides and other antimicrobial agents. In the in vitro activity azithromycin has a potent activity against both gram-positive and gram-negative bacteria, but against the latter organisms the other macrolides have little potent activity. The in vitro results demonstrate that azithromycin has a high potent activity against respiratory pathogens such as erythromycin-susceptible staphylococci, streptococci (except for organisms possessing erm gene), H, influenzae, B. prtussis, Legionella spp., M. pneumoniae, C. trachomatis. In the pharmacokinetic of azithromycin in mice, it is found that its half life is longer than the other macrolides and the concentration of it in polymorphonuclear leukocytes and lung tissues is higher than the other macrolides. These specific properties in the pharmacodynamic and the in vitro antibacterial activities reflect the therapeutic efficacies in experimental infection models. Finally, the therapeutic efficacies of azithromycin in various infection models better than those of the other macrolides. PMID- 12572088 TI - [Pharmacokinetics and tissue distribution of azithromycin in humans]. AB - Azithromycin(AZM) is a new macrolides antibiotic developed by Pfizer Inc. of the US. It has a 15-membered ring structure obtainable by introducing methyl substituted nitrogen into a 14-membered ring lactone of erythromycin(EM). An effective drug concentration is sustained long owning to its superiority in tissue distribution and long half life(T1/2). AZM is also demonstrated to be twice to three times more powerful than the existing macrolides owning to its reinforced antimicrobial activity against Gram-negative microorganisms including influenza virus. AZM is already used in clinical practice in foreign countries and its excellent clinical performance has been demonstrated by the data. As for the domestic basic and clinical studies of AZM, Phase I study was initiated in 1991 following completion of nonclinical studies, and general clinical studies, a dose-setting study and Phase III controlled studies were conducted between 1992 and 1994. Usefulness of AZM is substantiated by the data from those studies. PMID- 12572089 TI - [Transport to an infected site of azithromycin by phagocyte cells]. AB - Azithromycin is a new macrolide antibiotic developed by Pfizer. This compound has a 15 ring structure formed by adding a methyl-nitrogen to the 14-member lactone ring of erythromycin. Azithromycin has acid stability and significant improved activity against gram negative bacteria compared to other macrolides. Further, sustained high tissue levels of azithromycin have been demonstrated clinically and in basic research. There has been particular interest in the phagocyte delivery system of azithromycin to the site of infection. The mechanism is characterized by the intake of azithromycin by phagocytic cells which release the antibiotic at the site of infection. This report describes the mechanism of sustained high tissue levels by summarizing the data of Japanese and western clinical trials and research. PMID- 12572090 TI - [Clinical evaluation of azithromycin for respiratory infectious diseases]. AB - Azithromycin(AZM), an azalide macrolide with a 15-membered ring, is superior in cell/tissue penetration. Sustained pulmonary tissue concentrations are maintained which are higher than various MICs for major respiratory pathogens. Much research and clinical investigation has been done to evaluate the clinical efficacy of azithromycin abroad. In Japan, an optimal dose-finding pneumonia study was started in January, 1993, and a controlled comparative pneumonia study with clarithromycin was started in December of the same year. Patient compliance was excellent with the 3-day once daily regimen of AZM(500 mg). Azithromycin has been demonstrated to be highly clinically useful based on high efficacy (including the elderly population) and an established safety profile. PMID- 12572091 TI - [Clinical studies of azithromycin, a new macrolide antibiotic, for infections in the field of surgery]. AB - The efficacy of a newly developed macrolide antibiotic, azithromycin, for infections in the field of surgery, was investigated clinically by means of collaborative studies conducted in 17 major institutes and their affiliated hospitals throughout Japan. The following results were obtained. Clinical assessment: Azithromycin was administered at a dose of 250 mg or 500 mg once a day for 3 days. Clinical efficacy was evaluated in 170 patients. These subjects consisted of 81 with superficial purulent diseases, 12 with mastitis, 25 with periproctal abscess, 42 with superficial secondary infection due to trauma, burn and operative wound, 5 with cholecystitis or cholangitis, and 5 with other infections. The clinical efficacy rate was 96.3% (78/81) for superficial purulent diseases, 83.3% (10/12) for mastitis, 84.0%(21/25) for periproctal abscess, and 76.2%(32/42) for superficial secondary infection due to trauma, burn and operative wound. The overall clinical efficacy rate was 88.8%(151/170) respectively. The bacteriological eradication rate was 87.9%(116/132) for gram positive bacteria, 85.0%(34/40) for gram-negative bacteria, and 100%(63/63) for anaerobic strains of casual bacteria, which were isolated from 140 patients. The overall bacteriological eradication rate was 90.6%(213/235) respectively. Adverse effects were observed in 6 of 170 patients in whom they were evaluated. They consisted of gastrointestinal symptoms in 5 patients and exanthema in 1. Abnormal changes in clinical laboratory test values were observed in 5 patients, and consisted of eosinophilia in 1, elevations of S-GOT and S-GPT in 1, elevations of S-GOT, S-GPT and gamma-GTP in 1, elevation of S-GPT in 1, and elevations of AL-P and gamma-GTP in 1. These results suggest that azithromycin is very useful for surgical infections in the field of surgery. PMID- 12572092 TI - [Summary of the clinical studies with azithromycin in the pediatric fields]. AB - The clinical studies with azithromycin fine granules and capsules were conducted during the period from March 1993 to October 1994. Cmax's in 16 patients who received 10 mg/kg fine granules, were 0.29 +/- 0.24 microgram/ml, T1/2's were 42.0 +/- 11.8 hours, and AUC 0 approximately infinity's were 10.72 +/- 5.00 micrograms.hr/ml. The clinical results for azithromycin fine granule and capsules 10 mg/kg once daily for 3 days are as follows. The efficacy rate of fine granules, combining both "Excellent" and "Good", for pneumoniae where causative pathogenes were identified, was 95.3%, and for those which had failed to respond to previous chemotherapies, was 94.6%, respectively. The efficacy rate of capsules for 3 to 5 days was 100% in 40 cases where causative pathogenes were identified. Adverse reactions were found in 2.5%(fine granules) and in 5.4%(capsules) in cases eligible for evaluation. Abnormal changes in laboratory test were as follows: decrease of WBC by 5.6%(fine granules) and 9.3%(capsules) and increase in eosinophils by 7.1%(fine granules) and 11.4% (capsules). 59.8% of the patients claimed that the azithromycin 10% fine granules product was "easy to take". The result of a questionnaire on parents' demand on the improvement of antibiotics, showed that most concern was on the drug frequency(preferably once or twice daily) and the drug administering period(preferably short: 3 days). With regard to the efficacy, safety and compliance, it can be concluded that Azithromycin is one of the useful therapeutic regimens in the treatment of pediatric infections. PMID- 12572093 TI - [The clinical study of azithromycin in otolaryngologic infection]. AB - In this study, we evaluated the efficacy of azithromycin in otolaryngologic infections and tissue penetration. Azithromycin is a new macrolide developed by Pfizer Pharmaceuticals. Azithromycin maintained sustained high tissue levels compared with serum levels following small doses administered over a short period of time. Furthermore, excellent efficacy was achieved with the 3 day regimen of azithromycin, comparable to 10 day regimens of other antibiotics. In summary, azithromycin was demonstrated to be a highly useful antibiotic in infections of ear, nose, and throat. PMID- 12572094 TI - [Azithromycin in the field of dermatology]. AB - Azithromycin (AZM) is a new macrolides antibiotic that has a 15-membered ring structure obtained by introducing methyl-substituted nitrogen into a 14-membered ring lactone of erythromycin(EM). This article reviewed and summarized the clinical studies in the treatment of skin and skin structure infections conducted in Japan and abroad of AZM. PMID- 12572096 TI - [Clinical safety of azithromycin]. AB - The safety of azithromycin has been assessed in the clinical trial in Japan. Among the patients treated with azithromycin, side effects were recorded in 82(4.35%) of 1,886 adults and in 22(3.03%) of 726 children. The most common side effects in both groups were diarrhea, abdominal pain, and other gastrointestinal symptoms. All side effects were classed as mild or moderate. Laboratory abnormalities were recorded in 125(7.75%) of 1,279 adults and 85(19.23%) of 442 children, which included eosinophilia, neutropenia, and increases of GOT and GPT, and so on. All laboratory abnormalities were not severe and transient. Overall, azithromycin was well tolerated and can be safely used to treat bacterial infections in patients of all ages. PMID- 12572095 TI - [Azithromycin in dentistry and oral surgery]. AB - Azithromycin has adequate tissue levels for 7 days with a 3 day treatment regimen. The antibacterial activity is excellent. In dental infections, recurrence of infection happens when antibiotics cause a temporary improvement in symptoms but the etiologic agent is not eradicated. Azithromycin produces a "focused" cure and is an appropriate antibiotic for these infections. For this reason, the antibiotic was approved. PMID- 12572097 TI - [Lasers in dentistry 8. Use of lasers in oral and maxillofacial surgery]. AB - Since the 1970's lasers are used in oral and maxillofacial surgery. The effect of a laser on tissue is determined by the wavelength of the laserlight and the tissue specific absorbtion. Lasers are used for evaporation, excision and coagulation of tissue. The CO2-laser, the Nd:YAG-laser and the Argonlaser are used for these purposes. Light and laserlight are used for the diagnosis of mucosal lesions. By using different excitation wavelength autofluorescence of lesions can be detected and analysed. An artificial neural network can be used to analyse these data. Photodynamic Therapy (PDT) is a cancer therapy based on the more or less specific absorption of a drug in tumour, that can be activated with light of a specific wavelength. Activation of this drug causes tumour destruction due to the formation of oxyginradicals. PDT is limited by the penetration of the activating light and is therefore only suitable for the treatment of superficial tumours. PMID- 12572098 TI - [Implant-retained overdentures compared with complete dentures with or without preprosthetic surgery. Satisfaction and subjective chewing capabilities]. AB - The aim of this prospective randomized clinical trial was to evaluate and compare a set of clinical items and satisfaction of a group of edentulous patients during a 5-year follow-up. They were treated according to one of the following modalities: 61 patients with a mandibular overdenture on two implants (IMP group), 60 patients with a conventional complete denture (VP-group) and 28 patients with a complete denture after preprosthetic surgery (MVP-group). It can be concluded that endosseous implants, serving as retention for a mandibular overdenture, have a high survival rate after 5 years of follow-up (93%). The mean satisfaction score of the VP-group was lower than of the IMP-group. The mean satisfaction score of the MVP-group is lower than of the IMP-group. PMID- 12572099 TI - [Complaints of patients concerning obligation to inform and consent requirements]. AB - Since April 1995, patients' right to information and the duty of the operator to obtain their consent are legally established in the 'Medical Treatment Contract Act'. The aim of this study was to make an inventory of the number and type of information- and consent-complaints of Dutch dental patients, archived by the Dutch Dental Association in the years 1987-2000. By means of a registration form, complaints were analyzed and categorized. The results show that the number of complaints is slightly decreasing since the introduction of the 'Medical Treatment Contract Act'. However, complaints about lack of information on the treatment and consequences of treatment significantly increased. 60% of all complaints officially dealt with is (partly) substantiated. Redressment of done injustice, sometimes combined with a warning, is the sanction most often imposed. Although a substantial minority of the complaints is not substantiated, communication problems between dentist and patient do also seem to play an important role in those complaints. It is therefore necessary that dentists will provide comprehensible information to their patients. PMID- 12572100 TI - [Effects of low-intensity ultrasound on bone. Perspectives for dentistry?]. AB - Low-intensity ultrasound is frequently used for non-invasive diagnostic purposes. However, low intensity ultrasound can also be used as a therapeutical agent. It has been concluded from animal experiments that it significantly stimulates the growth of bone. In clinical trials an accelerated healing of fracture has been found. Future research has to define the role in dentistry and the final therapeutical value of low intensity ultrasound. PMID- 12572101 TI - [Guidelines for endocarditis prevention revised by the Netherlands Heart Foundation]. AB - The risk of endocarditis developing as the consequence of a bacteremia caused by a health care procedure in a patient with a predisposing heart disease is small. The administration of antibiotics before such a procedure is thought to reduce this risk still further. However, the protective effect of this preventive measure has never been demonstrated in a prospective randomized study. With certainty it can be said though, that it is not 100% efficacious and reports on prophylaxis failure have appeared ever since the introduction of this preventive measure. The revised guidelines are as far as possible based on published data. Where data are lacking, experts opinion was followed. The guidelines are not exhaustive and do not cover each and every circumstance in which prophylaxis could be warrantable. The purpose of the guidelines is to give the clinician something to go by in the most frequently occurring health care procedures. Not everyone will agree with the choices made by the commission in areas where data are lacking. In these areas, local directives may differ from the published directives. For a made to order advice on prophylaxis one should turn to an expert on endocarditis. PMID- 12572102 TI - [Firework injuries to the face]. PMID- 12572103 TI - [Asbestos and mesothelioma]. PMID- 12572104 TI - Take your children to work. PMID- 12572106 TI - Stewards of the environment: dentists and amalgam separators. PMID- 12572105 TI - Use of a tooth-colored post for anterior restorations. AB - The restoration of endodontically treated teeth is still a challenge to the general dentist. Extensive caries or traumatic fracture may result in insufficient coronal tooth structure remaining to support a core. The introduction of fiber post systems in the 1990s has provided dentists with new options for the restoration of the pulpless tooth. This clinical article demonstrates the use of a fiber post and a composite buildup material. PMID- 12572107 TI - It's only money. Part four: tax-deferred investing. PMID- 12572108 TI - Give kids a smile. PMID- 12572109 TI - Living on the edge. PMID- 12572110 TI - Esthetics and ethics in orthodontics. PMID- 12572111 TI - Joint replacement surgery and joint stability. PMID- 12572112 TI - The wire-reinforced banded herbst appliance. PMID- 12572113 TI - Treatment initiation: the beginning of great orthodontic treatment. Part II. PMID- 12572114 TI - To close or not to close. PMID- 12572115 TI - [Certain principles of Mars mission crew life and the organization of the performance]. AB - Article is devoted to some new aspects of biomedical support and countermeasures during a manned Mars mission. Significant differences in crew--Mission control interaction in a Martian and orbital flight are described. Special attention is paid to the problem of the Martian crew autonomy and decision-making. The authors give some practical recommendations for improvement of the space flight medical support system existing in Russia to meet the needs of a Martian expedition. PMID- 12572116 TI - [The problem of developing a lettuce greenhouse for the International space station and future interplanetary missions]. AB - In order to evaluate the effects of gravity on growing plants, we conducted ground-based long-term experiments with dwarf wheat (cultivar "Apogee USU") and Chinese cabbage (cultivar "Khibinskaja"). The test crop had been grown in overhead position with HPS lamp below the root module so that gravity and light gradients were in opposite direction. Plants of the control crop grew in normal position under the same lamp. Both crops were grown on porous metallic membranes with stable--1 kPa water potential on the surface. Results from these studies allowed us to examine the significant differences in growth and development of the plants as well as the root systems in relation to the gravity force. Nevertheless, the experiments in greenhouse Svet aboard the Mir space station proved that it is possible to compensate the effects of weightlessness on higher plants by manipulating gradients of environmental parameters (i.e. photon flux, water potential in the root zone, etc.). Even in ground studies Svet productivity averaged no more than 14 gm of fresh salad biomass per a day. This does not provide a sufficient supplement nutrients to the ISS crew. A cylindrical design of a space plant growth facility (SPGF) allows for maximal productivity under very tight energy and volume limitations onboard the ISS and a number of operational advantages. PMID- 12572117 TI - [Adrenoreactivity as a criterion for the evaluation of some professionally important qualities of the human operator]. AB - Adrenoreactivity parameter (beta-ARM, one of the characteristics of individual sympathoadrenal system (SAS), can enhance the system of professional psycho logical screening (PPS). The parameter is determined with a new technique of quantifying changes in osmoresistance of erythrocytes in the presence of adrenoblocker. The normal beta-ARM range lies within 2.0-20.0 conditional units with a rise up to 60 cond. units in consequence of regular increases in the SAS activity. To accept or reject beta-ARM as a PPS criterion, data of a series of three investigations were analyzed in attempt to correlate individual psychophysiological characteristics, operator's efficiency, cardiovascular reactions, and beta-ARM. In the first investigation, 30 male volunteers imitated operator's duties which included two-dimensional compensatory tracking with simultaneous choice between two alternatives on a training simulator. In the second investigation, 39 first-year students of a flight school were tested with the use of an automated psychological assessment system at the end of the first semester. In the third investigation, four navigators and 3 qualified test-pilots performed two-compensatory tracking using standard hand controls following exposure to radial aerobatic g-loads in and w/o anti-g suits. It was found that beta-ARM within the range of 10-20 cond. units is a criterion of good job efficiency. beta-ARM below 10 cond. units attests high adrenoreactivity only in stable conditions of work and with good skill in a particular operation. In contingency high reactivity to catecholamines may lead to loss of concentration, deterioration of temporary memory and job efficiency. beta-ARM above 20 cond. units may be determined at high operator's performance; however, during screening the risk of stress-related diseases should be taken into consideration, too. PMID- 12572118 TI - [Relationship of insulin dependent metabolic disorders to efficiency of intensive operator's work]. AB - The purpose was to state correlation between insulin-dependent metabolic disorders and efficiency of intensive operator's work. The investigation included 12-hr mission on a flight simulator performed by 50 normal (aged 23-36) flight qualified pilots. Increase in the number of erroneous actions was in direct correlation with insulin (r = 0.74, p < 0.01) and in reverse correlation with glucose incretion (r = -0.594, p < 0.01) and STH (r = -0.90, p < 0.006). Metabolic tests (glucose and insulin) showed that psychoemotional loading due to the intensive operator's duties led to early fatigue and sharp straining of tissue structures in people with dysregulatory disorders in insulin metabolism. The psychoemotional loading may also provoke dysregulatory disorders and development of insulin-dependent disturbances. PMID- 12572119 TI - [The effect of gravity on blood volumes in the main cervical arteries in healthy humans]. AB - The study of the effect of gravity on blood volumes in the main cervical arteries was conducted with voluntary participation of 50 normal men and women. Blood volumes were determined with the ultrasound technique. The circulatory effect of gravity was simulated by administration of the active standing test. Circulating volumes in the common carotid, internal carotid and vertebral arteries in the supine position were 464, 342 and 110 ml/min, respectively and decreased significantly up to 352, 262 and 84 ml/min, respectively in the standing position. The orthostatic reduction in the volume of blood circulating along the vertebral artery was proportional to that in the internal carotid artery. Specific contribution of changes in vascular capacity to the blood volume reduction was shown to be predominant. PMID- 12572120 TI - [Characteristics of the human thermal state and environment in long-term space flight]. AB - Summarized are data of many-year studies of thermal loading and thermal state of humans in different microclimates. Comparative analysis of various approaches to thermal monitoring of cosmonauts in space suits was performed with the conclusion about advantages of parotid thermometry. To set microclimate standards for long term space flight, a complex index of the environment cooling potential (ECP) based on the use of an electrical dynamic catathermometer has been proposed. A correlation between ECP values and cosmonaut's thermal sensations was elicited. For the most part, the proposed recommendations have been verified or introduced in the space crew medical care system. PMID- 12572121 TI - [Morphological changes in the adrenal gland of Japanese quail chicken hatched out in microgravity]. AB - The paper presents results of studying effects of spaceflight factors on the adrenal structure in Japanese quail hatchlings (Coturnix coturnix japonica) incubated and kept alive 4 to 5 days aboard the space station Mir. In comparison with the ground controls, cortical cells in the adrenals of space hatchlings contained copious fat drops, mitochondrias, and significantly dilated cisterns of the endoplasmic reticulum. Accumulation of fat drops evidences lowered cholesterol transformation and disturbed genesis of steroids. Cytoplasm of the medullar chromaffin cells was full of dark large granules; some of these cells also had enlarged mitochondrias and dilated endoplasmic reticulum cisterns. PMID- 12572122 TI - [Sensitivity and parameters of the otolith as a spatially distributed system]. AB - Comparison of the de Vries pendulum and spatially distributed otolith models allowed physical interpretation of the pendulum model parameters and determination of the influence of otolith structure on its mechanical sensitivity when otolith dynamics is represented by a parallel displacement relative to the receptor surface. Proof was obtained that in this case otolith dynamics is independent of total mass, and mechanic sensitivity is not proportional to size of the organ. Possible explanations of the effects of changed dynamics conditions on otolith parameters in the fish are discussed. PMID- 12572123 TI - [The bactericide effect of laser infrared radiation]. AB - The investigation had the purpose to study the lethal effect of laser infrared radiation (1220-1320 nm) on various genotypes of E. coli K-12 cells. Laser was produced by a parametric LiNbO3 light generator pumped up by the NdYAG second harmonic radiation. Results were compared with effects of X-ray radiation at 200 kV. The irradiated bacteria were cultivated in a solid full nutrient medium over 24 hrs at 37 degrees C. Bacteria spread in monolayer on the surface of "starvation" agar (4% agar-agar) were exposed to each type of radiation at room temperature. Survivability of the cells was determined by counting macrocolonies after 2 days of growth at 37 degrees C. Laser radiation at a wave-length of 1270 nm was found to have the highest lethal effect. This corresponds to one of the peaks of molecular oxygen consumption. Based on comparative analysis of the lethal effectiveness, the bacterial strains revealed equal relative sensitivity to the laser and X-ray radiation. Laser effectiveness was power dependent. Hence, proof has been exhibited that laser radiation at a wave-length of 1270 nm can be used as a bactericide means. The apparent advantage of the means over the photodynamic therapy is that no dyes are required. PMID- 12572124 TI - [Serum phospholipids and processes of cholesterol esterification in the European North]. AB - Lipid profile was studied in blood serum of 146 European northerns (trans-polar Nenets Autonomous district and Archangelsk). Monolayer chromatography was used to determine sphingomyelin (SP), phosphatidyl ethanolamine, phosphatidylcholine (PC), lisophosphatidylcholine, glycerophosphatides and phosphatide acides. The northerns were found to have very low concentrations of blood PC. At the same time, the NAD population have a little higher SP which may play an adaptive role in terms of the cell membrane function. The transpolar natives appear to possess an adaptive mechanism of increasing total cholesterol and reducing cholesterol of high-density lipoproteins, and activation of cholesterol esterification resulting in decrease in the free cholesterol fraction in serum. PMID- 12572125 TI - [Correlation between empirical and statistical indices of cardiovascular deconditioning in response to orthostatic exposures]. AB - Data of heart rate and blood pressure from 26 astronauts during orthostatic testing before and after space flight and 15 healthy men during orthostatic testing before and after 10-days head-down tilt (HDT) were used to correlate two integral indices of cardiovascular deconditioning. The first, very simple index, was used earlier by Bungo M. W. et al. in order to ascertain the efficiency of peroral body dehydratation as a measure to prevent orthostatic intolerance in astronauts after space flight. This index sums up HR and BP variations from one orthosatic probe to another. The choice of the another index was based on our investigation of HDT effects on the healthy human cardiovascular system. This index was determined as a difference between linear discriminant function values separating distinctly groups of men with normal and lowered orthostatic tolerance. Analysis of regression showed a high correlation between these indices: 0.71 for the group of astronauts and 0.88 for the group of healthy men. The linear regression equations allowed calculate value of one index from value of the other with an admissible standard error. PMID- 12572126 TI - [Microvzor-2: a system for automated dry blood smear analysis]. AB - Upgraded system Microvzor-2 for investigation of blood morphology in space flight is composed of soft- and hardware for dry smear image analysis. Ground-based testing showed that it could be utilized to investigate a broad spectrum of blood parameters in space flight, including erythrocytes, hemoglobin content in erythrocytes, volumetric erythrocyte distribution, diagnostics of anisocytosis, and poikilocytosis, leukocytes and leukocyte count. Analyzed are smears prepared from a fixed volume of finger blood. The process consists of smear scanning, input of the vision field images in the spacecraft computer and ensuing automated identification and counting. Information is stored in a dedicated database. Results of analysis are displayed as forms and cell galleries. PMID- 12572127 TI - [Microbial destruction of harmful organic contaminants inherent to atmosphere of living space]. AB - The investigation into the microbial destruction of harmful organic contaminants in the air of habitable rooms was performed with the assumption that microorganisms have labile metabolism and synthesize inducible enzymes involved in transformation of organics compounds. Based on our observations, Methylobacterium D-08 and Arthrobacter pastens are able to synthesize inducible enzymes and transform ethanol, acetic acid, acetone, and aldehyde. Association of these microorganisms does not reveal acute antagonism in case of co-cultivation on mineral substrates with carbon provided by ethanol, acidic acid, acetone, and acetate aldehyde. The most rapid growth of microorganisms was observed at 28 degrees C and pH 7. Determination of the effects of different concentrations of harmful organics on carbon dioxide production by the microbial association showed that the threshold concentration of acetate aldehyde was close to 2 mg/l and acetone--4 mg/l. The transformation activity was not reduced even at the ethanol and acidic acid concentrations of 80 and 100 mg/l, respectively. The association of A. pastens and M.D-08 oxidized ethanol, acidic acid, acetone and acetate aldehyde to carbon dioxide and water up to 90%. PMID- 12572128 TI - [Application of the strategic management approaches to implementation of space technologies in health services by the example of telemedicine]. AB - Telemedicine serves as an argument for application of the strategic management technologies. A telemedicine implementation strategy has been developed to be introduced in the Russian health services. PMID- 12572129 TI - [Tissue respiration of the liver rats irradiated with super low doses of incorporated radionuclides of caesium]. PMID- 12572130 TI - [Do we dare to investigate parental love and ability to take care of their children?]. PMID- 12572131 TI - [From evidence to clinical recommendations: all together (as in the USA) or a process (as here)?]. PMID- 12572132 TI - [Children of mentally retarded mothers--an inventory. Small group at risk with considerable need of assistance during an insecure childhood]. AB - Children born to mentally retarded mothers usually have normal intelligence. Their fathers often suffer from psychosocial problems. These children often grow up in an environment of insecurity and understimulation. How many are there? In 1995 an inventory was made in Skaraborg county (275,000 inhabitants), Sweden, based on the mothers' registration with the county authorities responsible for supporting people with mental retardation and on the general population register. A prevalence of 1.4 per thousand was found. Reasons for a higher true prevalence are discussed. The majority of the mothers had mild mental retardation. The rate of childbearing among mentally retarded women fell from 30 to 15 percent over the last three decades up to 1995. The prevalence for women with mental retardation was at least 1.1 percent. PMID- 12572133 TI - [Ambitious American guidelines for control of risk factors in diabetes. Was the aim put too high?]. PMID- 12572134 TI - [Screening for postmenopausal osteoporosis. Women with distal radius fractures should be evaluated for bone density]. AB - In a case-control study at two country hospitals in Sweden, 110 consecutive postmenopausal women (age 50-75) with distal radius fracture were examined with the DXA-technique, on the injured radius, lumbar spine, and the right hip within three weeks after the fracture occurrence. Data was compared with 55 age-matched controls from The Swedish Population Register. The incidence of osteoporosis according to WHO's definition (T-score < -2.5) at any measurement site was higher in the fracture group, 44 per cent compared with 27 per cent in the control group. The fracture group had 12 per cent lower bone mineral density in the distal radius compared with the control group. A higher rate of previous fractures was noted in the fracture group compared with the control group. The study reflects the situation in general health care, where osteoporosis is common in postmenopausal women with distal radius fracture. This patient group can easily be identified and is suitable to be diagnosed for osteoporosis using bone density measurement prior to a decision being reached with regard to any treatment. According to the guideline for medical treatment (T-score < -2.0 and fragile-fracture) as outlined by the Swedish Osteoporosis Society, 75 per cent of postmenopausal patients with forearm fracture should be considered for such treatment. PMID- 12572135 TI - [Economic assessment based on a clinical study of risedronate. Fracture prevention in elderly women with osteoporosis is cost-effective]. AB - Based on a clinical study we assessed the cost-effectiveness of treating elderly osteoporotic women with the biphosphonate Risedronat in a Swedish environment. Costs, health effects and risk of fractures were taken from previously published sources. Health effects were measured as Quality adjusted life-years (QALYs) and life-years gained. Results were presented with and without costs of added life years. The cost-effectiveness was investigated for two patient groups. The first group consisted of elderly osteoporotic women and the second group, which was a subgroup of the first, consisted of elderly osteoporotic women with previous vertebral fractures. The cost per gained QALY, estimated at SEK 277,209 (osteoporotic women) and SEK 16,877 (osteoporotic women with a previous vertebral fracture) respectively, were in both cases below the threshold value of 520,000 SEK, indicating that the treatment was cost-effective in both patient groups. In a sensitivity analysis the cost-effectiveness for the group of osteoporotic women with previous vertebral fractures showed to be stable for variation in the parameters while the group of osteoporotic women showed to be more sensitive for variation in some of the parameters. PMID- 12572136 TI - [Antiepileptic treatment nearly six times more expensive since 1990]. AB - There is a six-fold increase in the costs for antiepileptic drug prescriptions in Sweden from 1990 to 2000. This is mainly caused by a gradual increase in the prescribing of new and more expensive drugs, since the total use of antiepileptic drugs, expressed as defined daily doses per 1,000 inhabitants, is almost unchanged during the same time period. A marked variation in the prescribing of new antiepileptic drugs between different counties in Sweden, suggests that the role of the new compounds is still unclear. The cost effectiveness of new antiepileptic drugs in relation to the older drugs requires further evaluation. PMID- 12572137 TI - [Interesting work is the most important factor when choosing a specialty. Results of a questionnaire among medical students and interns]. PMID- 12572138 TI - ["Beginning of human life is a chimera--and the death is inevitable"]. PMID- 12572139 TI - [A prescription worth following: "Be clever! choose living not only working!]. PMID- 12572140 TI - [Cross-disciplinary use of medical records is good for cooperation within health care]. PMID- 12572141 TI - [Asylum seeking persons and refugees should have same right to health care as residents]. PMID- 12572142 TI - [Different frames of references make the exchange of ideas more difficult]. PMID- 12572143 TI - [A VIP lane for consumers of insurance companies to get certificates?]. PMID- 12572144 TI - [Which arguments may be used in the debate about circumcision?]. PMID- 12572145 TI - [Final reply to Yngve Hofvander: Jewish circumcision is basically a religious issue not a medical one]. PMID- 12572146 TI - [An excellent biography of a pediatrician who became a professor of pediatrics and medical counselor]. PMID- 12572147 TI - Pregnancy disability ... what do I do? PMID- 12572148 TI - Reimbursement from auto no-fault insurers. PMID- 12572150 TI - Dr. Sherill Behnke, Central District trustee. Interview by Jeff Mertens. PMID- 12572149 TI - Protect yourself against liability due to employment practices. PMID- 12572151 TI - Periradicular radiolucency in the mandible. Lateral periodontal cyst (LPC) or odontogenic keratocyst (OKC). PMID- 12572152 TI - Expanded alternative dental assisting program unveiled. PMID- 12572153 TI - We answer your HIPAA questions--Part One. PMID- 12572154 TI - Create a powerful vision and take off! PMID- 12572155 TI - The CDC releases new hand hygiene guidelines. PMID- 12572156 TI - British study suggests tooth whiteners may also prevent staining. PMID- 12572157 TI - Study will utilize patients' own cells in attempt to correct oral cosmetic problems. PMID- 12572158 TI - New drugs show promise in attacking HIV. PMID- 12572159 TI - Trends in dental hygiene. Shaping the way dental practitioners do business. PMID- 12572160 TI - Aesthetic breakthrough. Soft tissue lasers. PMID- 12572161 TI - SensiStat. A new saliva-based composition for simple and effective treatment of dentinal sensitivity pain. PMID- 12572162 TI - Bleaching for various complexions. PMID- 12572163 TI - Innovations in endodontic instruments and techniques: how they simplify treatment. PMID- 12572164 TI - High-quality panoramic radiographs: achieving the ideal. PMID- 12572165 TI - Clinical use of a new metal-free restorative technology: case reports. PMID- 12572167 TI - Insurance freedom. Is there such a thing? PMID- 12572166 TI - Brain injury in sports related to trauma to the lower jaw. PMID- 12572168 TI - Dental computing, some small steps: working smarter is easier and saves money. PMID- 12572169 TI - Power toothbrushes: home care in high gear. PMID- 12572171 TI - Dental lasers. PMID- 12572170 TI - At-home tooth whitening. PMID- 12572172 TI - Helping patients listen to their mouths. PMID- 12572173 TI - Debating occlusion and evolution. PMID- 12572174 TI - Debating occlusion and evolution. PMID- 12572175 TI - Debating occlusion and evolution. PMID- 12572176 TI - Debating occlusion and evolution. PMID- 12572177 TI - Drugs and herbal remedies that affect blood clotting. PMID- 12572178 TI - In whose interest? PMID- 12572179 TI - Esthetic treatment of the maxillary anterior. PMID- 12572180 TI - Comparison of computer-assisted brush biopsy results with follow up scalpel biopsy and histology. AB - Oral brush biopsy results were compared with scalpel biopsy and histology to determine the positive predictive value of an abnormal brush biopsy finding. Of 243 patients with abnormal brush biopsies, 93 proved positive for dysplasia (79) or carcinoma (14) and 150 were negative for either dysplasia or carcinoma. Therefore, the positive predictive value of an abnormal brush biopsy was 38% (93/243). By using the oral brush biopsy, dentists can inform their patients that abnormal findings have a strong positive predictive value for dysplasia or carcinoma and therefore require follow-up confirmation by scalpel biopsy. PMID- 12572181 TI - The asthma patient and dental management. AB - Asthma is increasing worldwide at a dramatic rate. The way the medical community defines asthma is changing, from considering it solely a respiratory disease to recognizing it as a more complex inflammatory disease process. This means that the treatment of asthma is becoming multifocal, depending upon the varied symptoms displayed. Reflecting this change is a shift to the use of corticosteroids in addition to the more traditional pulmonary medications. Dentists should be aware of the newer classifications and treatment modalities. There needs to be an understanding of the role these pharmaceutical agents play in the needs and treatment of our dental patients. Recent trends in the diagnosis and management of asthma are examined and recommendations for managing the asthmatic patient are provided. PMID- 12572182 TI - Dental management of patients with renal failure. AB - Renal disease has become increasingly more prevalent in our society. There are many more people undergoing dialysis treatment and kidney transplants than there were only a decade ago. The dental care of these patients can be complex, given the medications associated with the disease and the medical conditions that result from inadequately functioning kidneys. This article reviews the literature regarding the dental management of patients with acute and chronic renal failure. PMID- 12572183 TI - Ethnicity and oral health in community-dwelling older adults. AB - Epidemiological studies consistently have found differences in oral health status between white and non-white elders. This has been attributed primarily to lack of access to dental care, especially preventive services. The study reported here examined psychosocial factors in an attempt to explain these differences. PMID- 12572184 TI - A clinical study of patients with hypersensitive teeth. AB - Hypersensitive teeth have been a nemesis to patients in every dental practice. This clinical study was done to find a causal relationship of hypersensitive teeth and other organs of special senses, namely, sight, hearing, taste, smell, and touch. The analysis of the findings offers a basis for conclusion that dentition hypersensitivity and hypersensitivity of the special senses have a causal relationship. The dental clinician can provide the patient with an understanding and explanation of the cause(s) of hypersensitivity that will assist in a cooperative analysis of the symptoms and aid in the treatment to alleviate the pain. PMID- 12572185 TI - Management of dental patients with allergies to natural rubber latex. AB - Reports of serious allergic reaction to natural rubber latex (NRL) are not uncommon in dentistry. The prevalence of Type I allergies in high-risk patient groups can range as high as 17% for health care professionals and over 50% in patients with spina bifida. A Type IV allergy to NRL chemical additives, also known as allergic contact dermatitis, is common in health care professions. To safeguard patients and coworkers, dentists must learn to recognize patients and workers at risk for NRL allergies, identify the associated symptoms, implement preventive measures, and encourage thorough diagnostics and management. This article reviews the latest advances in NRL allergies, particularly as they apply in dentistry. PMID- 12572186 TI - Referring patients for stress-related disorders. AB - Stress and psychological disorders can have a profound effect on the etiology, duration, and outcome of nearly all physical conditions. They also can interfere with the dentist's diagnosis and treatment of patients, particularly those who suffer from chronic pain. Dentists should be able to identify the possibility of the presence of stress and psychological disorders in patients and to make an appropriate referral when it is indicated. PMID- 12572187 TI - Hereditary angioedema: a case report and literature review. AB - Hereditary angioedema is an autosomal dominant condition with a typical presentation of diffuse edematous, painless, and nonpitting swelling of the soft tissues. The disease manifests itself primarily in the extremities, face, airway, and abdominal viscera. Proper diagnosis and treatment are essential as this condition has the capacity of becoming life-threatening due to potential airway embarrassment. This case report demonstrates the complexity of diagnosing and treating hereditary angioedema. Hereditary angioedema often is overlooked on a differential diagnosis of patients suffering from diffuse swelling and/or abdominal discomfort. Nonetheless, the diagnosis of hereditary angioedema should be included when assessing patients who display the proper symptomology and familial history of the condition. PMID- 12572188 TI - The posterior superior alveolar injection technique: a report on technique variations and complications. AB - The posterior superior alveolar (PSA) injection technique has varied over time with respect to the depth and angle of penetration, the location for deposition of anesthetic agent, and the number of injections necessary to assure adequate anesthesia to the maxillary molars. Of the standard intraoral injections, the PSA carries with it the second highest risk for anesthesia complications. With changes in armamentarium and technique, the complication rates have declined and more often are associated with anatomical considerations with respect to neurovascular compromise and/or anesthetic solution. In this study, the PSA injection technique and complication histories were investigated. Seventeen variations to the technique are reported along with 12 injection complications. A historic injection complication classification system is presented along with a management strategy based upon a review of reported provider experiences and treatment suggestions. PMID- 12572189 TI - The triangle of safety: a modified posterior superior alveolar injection technique based on the anatomy of the PSA artery. AB - The posterior superior alveolar (PSA) injection technique is commonly used to anesthetize soft and hard tissues of the posterior maxilla. As with all injections, complications arise, including hematoma formation secondary to needle induced vascular trauma. In an attempt to develop a hemorrhage-free PSA injection technique, 361 infratemporal dissections were completed on human cadaver specimens. Three distribution patterns were identified for the external branch of the PSA artery. Regardless of distribution pattern, an anatomical Triangle of Safety was found superior to the maxillary second molar that was free of neurovascular tissues in more than 99% of individuals. Injection into this area appears to meet anesthetic needs while reducing the risk of hematoma formation. The combination of this anatomical triangle with newer anesthetic agents and computerized delivery systems holds promise for continued improvement of the PSA injection technique. PMID- 12572190 TI - Chronic diffuse yellow eruption. Pyostomatitis vegetans. PMID- 12572191 TI - Exophytic mass of buccal mucosa. Blastomycosis. PMID- 12572192 TI - A clinical overview of removable prostheses: 3. Principles of design for removable partial dentures. AB - Removable partial dentures (RPDs) should not be made for patients unless they are necessary. Most partial dentures have the potential to cause some damage to the teeth and supporting tissues, however well they are designed and constructed; the criteria for selecting such devices were described in a previous article. In general there is merit in, wherever possible, reducing tissue coverage as much as possible when RPDs are being planned. This article, the third in a series on the prescription of RPDs, discusses the design principles involved. PMID- 12572193 TI - Can rapid prototyping ever become a routine feature in general dental practice? AB - Rapid prototyping is a technique for the production of solid objects from computer models. The variety of rapid prototyping devices and materials that can be processed is increasing all the time. In this paper, the authors examine the potential for the application of current devices to general dental practice and speculate on developments that could make this technique an attractive prospect within the dental surgery. PMID- 12572194 TI - The removal of fractured post fragments in general dental practice using ultrasonic vibration. AB - This paper describes an evidence-based clinical procedure that is suitable for the removal of fractured metal post fragments using ultrasonic vibration in general dental practice, particularly when non-resin-based cements have been used. Fragments of posts cemented with resin-based cements are likely to be extremely difficult to remove. The use of non-resin-based cements is recommended for luting posts, as fragment or total post removal is much easier with these materials. PMID- 12572195 TI - Potential hazards of orthodontic treatment--what your patient should know. AB - Orthodontic treatment carries with it the risks of tissue damage, treatment failure and an increased predisposition to dental disorders. The dentist must be aware of these risks in order to help the patient make a fully informed choice whether to proceed with orthodontic treatment. This paper outlines the potential hazards and suggests how they may be avoided or minimized. PMID- 12572196 TI - A practical guide for patients undergoing exodontia following radiotherapy to the oral cavity. AB - Dental extractions or minor oral surgery in patients who have undergone radiation therapy for cancer in the head and neck carry the risk of one of the most serious and devastating complications of head and neck radiotherapy, that of osteoradionecrosis (ORN). A totally unified approach to dental extractions following radiotherapy is lacking. The role of the general dental practitioner in management of patients following radiotherapy is crucial. Contrary to clinical impression, the risk of ORN does not decrease with time. When contemplating exodontia or minor oral surgery in the irradiated patient, special consideration should be given to issues such as radiotherapy history, surgical assessment, surgical procedure and the role of antibiotics and hyperbaric oxygen. PMID- 12572197 TI - The impact of communication technology on dental services and dental education. AB - In this paper, the author reviews the scope, and probable future impact, of information technology in dentistry. PMID- 12572198 TI - Preserving the vital pulpin operative dentistry. PMID- 12572199 TI - Facial sinus of dental origin. PMID- 12572200 TI - Iron and vitamin B12 deficiency anaemia in a vegetarian. PMID- 12572201 TI - The management of gingival recession. PMID- 12572202 TI - Give kids a smile! PMID- 12572203 TI - Evidence links pulmonary infection, oral diseases. PMID- 12572204 TI - Questions about patient bankruptcies. PMID- 12572205 TI - Rising health care costs: a report. PMID- 12572206 TI - "Give kids a smile": your opportunity to shine! PMID- 12572208 TI - The joy of treating children: a day in the life of a pediatric dentist. PMID- 12572207 TI - Promoting oral health care for our youth. PMID- 12572209 TI - Treating the child patient: twelve tips to success! PMID- 12572210 TI - Good communication in pediatric dentistry. PMID- 12572211 TI - HIPAA vs. Michigan Law: what you should know. PMID- 12572212 TI - Recent developments in dentin bonding agents. PMID- 12572213 TI - Knock on effects. PMID- 12572214 TI - Anthrax alert? PMID- 12572215 TI - Life and death. Interview by Daniel Allen. PMID- 12572216 TI - Facing a challenge. PMID- 12572217 TI - Qualified to care. PMID- 12572218 TI - Sick and tired. PMID- 12572219 TI - Psychological and psychosocial effects of prostate cancer. AB - BACKGROUND: Prostate cancer is a life-threatening disease that is not curable but can be controlled through treatment. Current treatments can result in unpleasant and distressing side effects for patients. Little was known until the 1970s about the psychological and psychosocial effects of these treatments, but studies have now been conducted on patient identity, self-esteem, incontinence, impotence and social support. CONCLUSION: A diagnosis of prostate cancer can have detrimental psychological effects for patients. However, using behavioural and coping strategies with cancer patients brings about an improvement in symptoms, functioning and emotion, thus enhancing quality of life for patients and their families. PMID- 12572220 TI - Learning to nurse in care homes: student support. AB - An increasing number of care homes in the UK now provide placements for students undertaking pre-registration nursing programmes, enabling them to develop knowledge and skills in the continuing care of older people. In this article the authors describe an initiative to support nursing staff working in care homes and prepare them for their role in student education. The impetus for the initiative came from student feedback and evaluation of their placement experiences. These suggested that staff in some care homes were not fully prepared to help meet students' learning needs. Students' concerns about the quality of care in some homes also made clear the need for a forum in which the evidence base for practice in continuing care settings could be discussed and debated. The initiative combined the development of new roles with regular opportunities to share information and feedback. PMID- 12572221 TI - An overview of chronic fatigue syndrome. AB - This article provides an overview of the condition known as chronic fatigue syndrome, or myalgic encephalomyelitis. The author describes common symptoms and their treatment, and discusses a model of patient care, piloted in the community, which includes elements of a range of therapeutic strategies. PMID- 12572222 TI - CDE, SELECT committee study dental workforce development. PMID- 12572223 TI - Testing your diagnostic skills (#66). Case No. 1. Pemphigus vulgaris. PMID- 12572224 TI - Testing your diagnostics skills (#66). Case No. 2. Dentigerous cyst. PMID- 12572225 TI - ADA, FDA reach out to those in need. PMID- 12572226 TI - ACORN, UF-CD serve needy communities. PMID- 12572227 TI - Mission trips strengthen relations and benefit Third-world people. PMID- 12572229 TI - Pain at first dressing change after toenail avulsion: the experience of nurses, patients and an observer: 1. AB - OBJECTIVE: Ingrowing toenails are often treated surgically by nail avulsion, with paraffin tulle gauze applied to the toenail bed afterwards. In one large city's primary care trusts, community nurses reported that patients felt pain when the gauze was removed postoperatively. This study looked at patients' general pain experience, and their pain during the first dressing change after toenail avulsion. Patients' perceptions of their pain were investigated, along with nurses' experience of redressing toenail beds, their technique and their interaction with patients, especially in relation to pain prevention. METHOD: First an extensive literature review was conducted. Then a collective instrumental case study design was used. This allowed triangulation of data sources using patient and nurse interviews, and non-participant observation, from six cases recruited from the city's primary care trusts. Data were subjected to thematic content analysis, with two main categories identified. These were 'setting up of expectations' and 'a painful experience: but for whom?' RESULTS AND CONCLUSION: This study provides a unique description of the type of pain patients experience. It raises serious issues about the adequacy of nurses' management of pain, including assessment, documentation and advice on pain-relief strategies. It supports discontinuing the use of paraffin tulle gauze as the postoperative dressing, and raises ethical issues about changing treatments and lack of communication with secondary care. Finally, it identifies a gap in knowledge of the most suitable dressing product to apply immediately after surgery. A randomised controlled trial is needed to clarify these preliminary findings. PMID- 12572230 TI - Questions & answers. Doppler ultrasound for healed ulcers. PMID- 12572231 TI - The role of neutrophil apoptosis in influencing tissue repair. AB - Neutrophils and macrophages are involved in inflammation. Neutrophils debride the area of local contamination and then die (apoptosis) after being engulfed by macrophages. This is vital before the wound can move to the next stage of healing. PMID- 12572232 TI - Assessing the value of a leg ulcer education programme in Hong Kong. AB - Patients with leg ulcers require expert care, but nurses do not always receive full training and education on this subject. A short education programme devised for community nurses in Hong Kong resulted in significant gains in knowledge. PMID- 12572233 TI - Topical negative pressure (TNP): the evolution of a novel wound therapy. AB - Topical negative pressure is a novel non-pharmacological therapy that is now being adopted as a standard of care in wound care management programmes. This review assesses where and how it can be best used. PMID- 12572234 TI - Controversies in classifying and assessing grade I pressure ulcers. AB - There is more uncertainty on how to define a grade I pressure ulcer than any other type. The confusion will need to be dispelled and a consensus reached if a national grading system is to be implemented. This review explores the options. PMID- 12572235 TI - The use of polarised polychromatic non-coherent light alone as a therapy for venous leg ulceration. AB - OBJECTIVE: This study assessed the effectiveness of polarised, polychromatic, non coherent light therapy in the treatment of venous leg ulcers. Investigators in previous studies have advocated the use of light as an adjunct to other proven therapies or on its own. METHOD: This was a pilot prospective case-series study. We enrolled 25 patients with venous leg ulcers. All were treated with light only. Phototherapy (light therapy) treatments were given once a day over four weeks. RESULTS: All ulcers except one (99%) had a positive value for the change in healing area at the end of the four weeks. The total number of 73 leg ulcers recorded at the beginning of the study was reduced to 51 at the end of the four weeks (p < 0.01). The decrease in wound surface area following the treatment was statistically significant (mean: 57.15%; SD: 31.87%; p < 0.01). CONCLUSION: Polarised, polychromatic light therapy applied as a monotherapy was associated with positive healing rates in patients with venous leg ulcers. It is a simple and non-invasive treatment. However, a well-designed randomised controlled study is needed to confirm the efficacy of this form of phototherapy and to objectively evaluate recommendations for its routine use in clinical practice. PMID- 12572236 TI - Prevalence of steroid receptors and HER 2/neu in breast cancer biopsies of women living in Puerto Rico. AB - OBJECTIVE: The purpose of this study is to determine the prevalence rate of estrogen and progesterone receptors and HER 2/neu in the breast cancer biopsies analyzed in the Laboratory of Immunohistochemistry of the University of Puerto Rico School of Medicine in the year 2000. This data may serve as a reference point for future studies of the epidemiological aspects of breast cancer among women living in Puerto Rico. BACKGROUND: Determination of estrogen receptor (ER) and progesterone receptor (PR) on biopsy specimens of breast carcinoma prior to treatment is standard practice in the management of breast carcinoma. ER and PR are used to identify patients who are likely to respond to endocrine therapy. The prevalence of ER, PR and Her2/neu among USA women is 77%, 55% and 10-34%, respectively. One of the major clinical roles for testing HER 2/neu expression is to determine eligibility for treatment with Trastuzumab. METHODS: Retrospective analysis of 309 breast cancer biopsies was done. Paraffin embedded blocks of breast cancer tissue biopsies were received from different hospitals and Pathology Laboratories located throughout the island specifically for routine analysis of steroid receptor (ER/PR) and/or HER 2/neu expression. Immunostaining was performed in a Ventana Medical Systems automated instrument. RESULTS: Positive nuclear staining for ER and PR were seen in 65.9% (203/308) and 51.8% (159/307), respectively. In the HER2/neu test, 27.8% (46/165) gave a strong and complete membranous staining (score 3+). CONCLUSIONS: There is a lower prevalence of estrogen receptor in the breast cancer biopsies of women living in Puerto Rico than their USA counterparts, but similar prevalence of progesterone receptor status and HER 2/neu protein over expression. PMID- 12572238 TI - Acute respiratory distress syndrome in the trauma intensive care unit: the other view. AB - OBJECTIVE: To study the timeliness of the diagnosis of patients with acute respiratory distress syndrome (ARDS)/acute lung injury (ALI) at the Puerto Rico Trauma Center (PRTC) and to determine the overall 28 day mortality for ARDS during the study period. METHOD: A retrospective review of all admissions to the Trauma Intensive Care Unit (TICU) from August 2000 to August 2001 was done. Patients with the diagnosis of ARDS/ALI were selected, records examined, and clinical data obtained for analysis. FINDINGS: Of the 537 patient admitted to the PRTC, 236 patient were admitted to TICU. Of these, 17 patients were identified as having hypoxemic ratios below 200 and 13 patients were identified as having ARDS as established by the American-European Consensus Conference of 1994. Their mean age was 41 years, the main cause of ARDS was due to pulmonary contusion due to blunt chest trauma. The 28-day survival for this group was 43%. Subgroup analysis showed that there was 86% mortality when the polytraumatized patient developed sepsis. CONCLUSION: The majority of the cases of ARDS were correctly identified as such by caregivers at the time of diagnosis. Mortality as predicted by Injury Severity Score in our ARDS patients' correlates with overall mortality in our TICU. Mortality in this group is lower than that of reported literature, in sharp contrast to our medical ICU counterparts. PMID- 12572237 TI - Acute lung injury/acute respiratory distress syndrome: a need for education. AB - OBJECTIVE: To examine the timeliness of the diagnosis of patients with ALI/ARDS at the San Juan VA Medical Intensive Care Unit. We were also interested in determining the incidence and the overall 28-day mortality for ARDS during the study period. METHODS: Retrospective record review of all admissions to the San Juan Veterans Affairs Medical ICU during a two-year period (1997-1998). RESULTS: During the study period, 587 patients were admitted to the medical ICU. All had APACHE II scoring performed during their first 24 hours of admission. Twenty three patients were found to have an A-a gradient of 350 or less. However, two patients were later identified as having radiographic changes compatible with Congestive Heart Failure and were excluded from the study. The incidence of ALI/ARDS was found to be 3.6% at our institution. Of the 21 patients with ARDS, in only 4 the diagnosis of ARDS was documented on their charts. CONCLUSION: Our findings suggest that education in the recognition of ARDS should be aggressively done. In order to implement the recently published successful strategies in the mechanical ventilation of patients with ARDS, it should be first recognized. Our patients need it. PMID- 12572239 TI - [Cardiovascular center of Puerto Rico and the Caribbean: 100 years of operations]. PMID- 12572240 TI - Effects of a high molecular mass Convolvulus arvensis extract on tumor growth and angiogenesis. AB - BACKGROUND: Plant materials represent promising sources of anti-cancer agents. We developed and tested a novel extract from the ubiquitous plant Convolvulus arvensis. MATERIALS AND METHODS: Convolvulus arvensis components were extracted in boiling water, and small molecules were removed by high-pressure filtration. The extract's biological activity was assessed by measuring its effects on S-180 fibrosarcoma growth in Kun Ming mice and on heparin-induced angiogenesis in chick embryos. We also examined the extract's effects on lymphocytes ex vivo and tumor cell growth in vitro. RESULTS: The extract (primarily proteins and polysaccharides) inhibited tumor growth in a dose-dependent fashion when administered orally. At the highest dose tested, 200 mg/kg/day, tumor growth was inhibited by roughly seventy percent. Subcutaneous or intraperitoneal administration at 50 mg/kg/day also inhibited tumor growth by over seventy percent. The extract's acute LD50 in Kun Ming mice was 500 mg/kg/day when injected, indicating that tumor growth inhibition occurred at non-toxic doses. It inhibited angiogenesis in chick embryos, improved lymphocyte survival ex vivo, and enhanced yeast phagocytosis, but did not kill tumor cells in culture. CONCLUSION: High molecular mass extract deserves further study as an anti-cancer agent. PMID- 12572242 TI - [The need for training in gerontology and geriatrics among the staff providing services at a geriatric care institution]. AB - The purpose of this qualitative study was to explore and identify the basic training needs of nursing home staff, in terms of attitudes, knowledge and skills required to work effectively with geriatric patients. Three focus groups were performed, two groups of employees, and a group of elderly residents of the institution, in order to explore issues pertaining to the following topics: personal attributes required to work with geriatric patients, basic knowledge and skills needed to provide effective services. Group discussions were transcribed and themes were extracted through consensus reached by the investigators. Results indicated that the interviewed staff lack of formal preparation or continuing education in gerontology or geriatrics. Needs identified were the following: the aging process, caring behaviors, management of common health conditions, administration of medications, transference and mobility of residents, among others. Finding were use to design an educational program aimed in assisting nursing home staff in providing an effective service to their geriatric patients. PMID- 12572241 TI - Prevalence of primary and secondary resistant mutations to antiretroviral drug in a population of Puerto Rican infected with HIV. AB - INTRODUCTION: Several studies have reported increasing number of therapeutic failures with HAART in HIV-infected individuals. In order to assess the impact HIV antiretroviral resistance could have on treatment, we decided to determine the prevalence of primary and secondary antiretroviral resistant genotypes in a population of HIV-infected Puerto Ricans and compare the mutational distribution pattern with that reported in Europe and US. METHOD: In a total of 80 plasma samples from patients with detectable viral load of over 1,000 RNA copies/ml, the Trugene Visible Genetics HIV sequencing method was used to detect antiretroviral resistance mutations. RESULTS: We found 55 subjects (69%) with high level of resistance to ZDV in the reverse transcriptase gene and 46 subjects (58%) with high level of resistance to NFV in the protease gene. Mutation frequencies to the NRTI ranged in appearance from as high as 54% (i.e., M184V) in the studied subjects to a low of less than 5% (i.e., M184I and V75T). For the NNRTI the most common mutation was K103N in 40% of the subjects and found to confer cross resistance to NVP, DLV and EFV. Another concerning finding is the increasing trend of the frequency of primary and secondary resistant mutations from year 2000 to 20001. Nine (23%) of the total detected primary mutations, to either RTI or PI, showed an increase of at least 5% from one year to the other. Similarly, there were 6 (11%) secondary resistant mutations showing an increase of at least 5% during the two years studied. CONCLUSIONS: In two year period we detected a tendency to increase in primary and secondary HIV-resistant mutation in a population of HIV-infected Puerto Ricans. PMID- 12572243 TI - Prospective study using standardized methodology for antimicrobial susceptibility of gram-positive cocci isolated from the Puerto Rico Medical Center. AB - The Gram-positive cocci (GPC), Staphylococcus aureus, Streptococcus pneumoniae, Enterococcus faecalis and Enterococcus faecium, have become important causes of community and nosocomial-acquired infections. The prevalence of multiple resistant isolates to standard antimicrobial drugs has significantly increased over the past decades. Few prospective studies have been performed in Puerto Rico (PR) concerning the GPC antimicrobial susceptibilities pattern. The purpose of this study was to evaluate the in vitro susceptibility of GPC clinical isolates from PR to selected standard antibiotics and to the new antimicrobial agents, linezolid (LZ), quinupristin/dalfopristin (Q/D) and gemifloxacin (GM). The in vitro susceptibility utilizing disk diffusion and Etest methods to selected antibiotics was determined for a total of 429 isolates obtained during a period of 5 months from the Puerto Rico Medical Center Bacteriology Laboratory. The distribution of GPC collected was as follows: 213 S. aureus isolates, 162 E. faecalis, 16 E. faecium and 38 S. pneumoniae. The results of the susceptibility test demonstrated: 1) that in S. aureus, 100% of the isolates were susceptible to vancomycin (VAN), LZ and Q/D; 93% to GM; and 61% to methicillin/oxacillin; 2) in S. pneumoniae, 100% were susceptible to LN and GM; 87% to Q/D; and 53% to penicillin; 3) in E. faecalis, 99% were susceptible to ampicillin; 93% to LZ; 79% to GM; 78.6% to VAN; and 0% to Q/D. Sixty eight and 66% of the E. faecalis isolates were susceptible to gentamicin and streptomycin respectively; and 4) in E. faecium, 100% were susceptible to LZ; 94% to Q/D; 69% to GM; 37.5% to VAN and 20% to ampicillin. In E. faecium isolates, 50% and 31% were susceptible to gentamicin and streptomycin, respectively. Of the vancomycin resistant enterococci, 88.9% and 21% of E. faecium and faecalis showed VanA phenotypic resistance, respectively. These results show that there is a significant degree of antimicrobial resistance in GPC, including 38% methicillin resistance in S. aureus, a near 50% penicillin resistant S. pneumoniae, and a significant resistance of enterococcal species to VAN. The new agents, LZ, Q/D and GM, proved to be effective against both, S. aureus and S. pneumoniae. For E. faecium, both, LZ and Q/D were active, while for E. faecalis, only LZ showed consistent activity. PMID- 12572244 TI - Incidence of fetal echogenic intracardiac foci in a Hispanic population. AB - Echogenic intracardiac foci (EIF) are small areas of increased echogenicity inside the fetal ventricles. When isolated, they are considered to be a normal finding with prevalence differing among ethnic groups. It has been described as a weak marker for trisomy 21 and other chromosomal anomalies. Little information exists regarding the incidence of these foci among Hispanic fetuses. We examined prospectively 485 normal fetuses between 14 and 32 weeks of gestation from January through March 2001. Nine cases of isolated intracardiac foci were identified (incidence of 1.8%). None of these cases had a chromosomal anomaly. Our data shows a frequency in our population similar to that reported among Caucasians. PMID- 12572245 TI - [Common competencies for the prevention of violence among young people]. AB - The high prevalence of violence in children and youth has been a great concern among diverse sectors of our society. Considered as a complex public health problem, the Centers of Disease Control and Prevention (CDC) of Atlanta, Georgia, has financed local and national projects geared to its prevention. This work describes the process in which the Developing Centers of Youth Violence Prevention from the University of Puerto Rico and the University of Southern California collaborated in the development of core competencies for health professionals in youth violence prevention. This two Developing Centers are projects funded by the CDC. PMID- 12572246 TI - [Factors associated with the use of tobacco among students of the Medical Sciences School, University of Puerto Rico]. AB - The objective of this study is to evaluate associated factors related to cigarette smoking in students of the Medical Sciences Campus, University of Puerto Rico. One hundred eighty-nine students, registered in the summer session, participated in the study. A self-administered questionnaire was used to collect the information. For the data analysis descriptive statistics were used, as well as Pearson Chi Square and Mann-Whitney test. The results showed that 12.2% of the participants are smokers at the moment of the study. Most of the smokers stated that the main reason they smoke was for stress reduction and psychological addiction. A statistical association was encountered between the cigarette use and exposure of information (X2Fisher (1) = 4.65, p = 0.02). We conclude that the participants smoke as an adaptative response to the daily stressors that impact the academic setting. PMID- 12572247 TI - [Bioethical reflexions on cloning of human beings]. PMID- 12572248 TI - [Publications by authors affiliated with the School of Medicine, University of Puerto Rico: 1950-2000. III]. PMID- 12572250 TI - Autotransplantation of a permanent maxillary incisor. PMID- 12572251 TI - Titanium appliances for allergic patients. PMID- 12572252 TI - Custom-made rotational ligatures. PMID- 12572253 TI - Working with children: from compliance to collaboration. PMID- 12572254 TI - Clinical evaluation of bond failure rates with a new self-etching primer. PMID- 12572255 TI - 2002 JCO study of orthodontic diagnosis and treatment procedures. Part 3. More breakdowns of selected variables. PMID- 12572256 TI - Transitional office design: attracting an associate. PMID- 12572257 TI - An orthodontist's perspective on the use of habit appliances. PMID- 12572258 TI - Suffer the little children: fixed intraoral habit appliances for treating childhood thumbsucking habits: a critical review of the literature. AB - A critical review of the literature is presented covering the treatment of childhood thumbsucking habits using fixed intraoral habit appliances (hayrake, palatal crib). The habit appliances are classified into type and function. Data is tabulated for key references revealing the fragmented and distorted nature of the literature and its lack of consistency. A chronological approach is presented to confirm the confused and idiosyncratic character of the literature. Information is provided on the early work of Massler and Graber and the paradox of Mack, Korner and Reider. Haryett's seminal studies at the University of Alberta regarding aspects of the treatment used are critically reviewed. Reflections are presented on why Larsson's study, casting doubt on the wisdom of using habit appliances, continues to be ignored. The emergence of the Bluegrass Appliance is discussed in terms of its being a more humane appliance and the seeming reluctance of practitioners to apply it as a kinder form of appliance therapy. Information is reported on the pain and serious injuries inflicted on children by habit appliances. A comparison of the use of appliances in the USA is made with the UK, where fixed habit appliances are not popular. Concludes that fixed intraoral habit appliances are cruel and inflict pain and suffering on children out of all proportion to their necessity. Questions why these appliances continue to be used, implying that it could be a combination of financial inducement, professional insularity and the absence of concerted opposition from behavioural therapists. PMID- 12572259 TI - Orofacial myofunctional therapy in dysarthria: a study on speech intelligibility. AB - Various types of orofacial myofunctional disorders co-exist with speech problems in individuals with dysarthria. Controlled studies stating the efficacy of orofacial myofunctional therapy (OMT) in dysarthric individuals are very scant. The present study was undertaken to examine the efficacy of OMT in twelve patients diagnosed with mild to moderate dysarthria following right hemisphere brain damage. Pre-therapy assessment focused on existing orofacial myofunctional problems and speech intelligibility in the clients. The goals of OMT were to increase strength and mobility of buccal, facial, labial, and lingual musculature. No speech intervention was provided while OMT was in progress. Post therapy measures indicated significant improvements in the stated goals as well as in speech intelligibility for single words. Patients observed functional improvements in swallowing functions too. A high positive correlation was found between speech intelligibility and diadochokinetic rate. Clinical implications regarding use of OMT in dysarthria are discussed. PMID- 12572260 TI - Physiological effects of an 8-week mechanically aided resistance facial exercise program. AB - This paper describes a study that for the first time addresses the physiological effects of an 8-week mechanically aided facial exercise program, using the Facial Flex device (Facial Concepts, Inc., Blue Bell, PA) with four healthy individuals with no motor, speech, language, or hearing problems. For a variety of non-speech and speech tasks, upper and lower lip muscle activity (EMG) and upper and lower lip movements were recorded at two baseline sessions (separated by 1 week) and immediately after an 8-week training period. The results indicate that after the training period, all four subjects showed an increase in the number of task repetitions and the duration of isometric contraction using the Facial-Flex device with a fixed resistance (Linebaugh tests). However, with respect to physiological changes as related to the exercise program, the results were mixed. Only one subject showed the expected significant increase in normalized EMG activity. This response was mirrored in a significant overall increase in movement range and peak velocity after the 8-week training period. Regarding the other three subjects, one subject showed no systematic training effect at all, whereas the remaining two subjects showed a significant increase in movement duration. Non-speech and speech tasks were found to be clearly different in their overall physiological characteristics; speech related movements were found to be more clearly defined in terms of larger amplitudes, shorter durations, higher peak velocities, and less variable movement cycles. The apparent discrepancy between the results of the Linebaugh tests and the physiological measures on specific oro-motor tasks warrants some caution in drawing conclusions on changes in the oro-motor system based on general performance measures. Further studies with well-defined clinical populations are needed to assess the usefulness of this device as an aid in the treatment of speech disorders based on motor system impairments. PMID- 12572261 TI - [Time is our most important capital]. PMID- 12572262 TI - [When you cross your hands, which of the thumbs is on the top?]. PMID- 12572263 TI - [A serious chewing gum problem]. PMID- 12572264 TI - [The medical education of my dreams at the Faculty of Health Sciences' ReijoWaara Institute]. PMID- 12572265 TI - [Edema at Christmas time]. PMID- 12572266 TI - [The physician as an exercise consult?]. PMID- 12572267 TI - [There is time and will to travel]. PMID- 12572268 TI - [The petrified time]. PMID- 12572269 TI - [The living time--on the relativity of biological time]. PMID- 12572270 TI - [RNA--the original chemistry of life]. PMID- 12572271 TI - [The chance and necessity of life]. PMID- 12572272 TI - [Time is genetic success]. PMID- 12572273 TI - [Infections as catalysts of the evolution]. PMID- 12572274 TI - [On the Maid Finland, the Neandertal human and shortly on the diet of Otz. Time traveling with the help of a molecule]. PMID- 12572275 TI - [The squirrel wheel of cultural evolution]. PMID- 12572276 TI - [Evolution and the language]. PMID- 12572277 TI - [The effects of Christmas on health]. PMID- 12572278 TI - [Are child psychiatrists doing the wrong things?]. PMID- 12572279 TI - [Duodecim's first Cultural Award to Kari Heiskanen]. PMID- 12572280 TI - [Lymphatic growth factors play a part in metastasis formation]. PMID- 12572282 TI - [Appendicitis in children--the number of unnecessary operations declining]. PMID- 12572284 TI - [An aggressive exacerbation of herpes virus infection]. PMID- 12572285 TI - [Experiences about electronic consultations]. PMID- 12572286 TI - [Thioridazine and the warning about prolonged QT time]. PMID- 12572287 TI - [Treatment of stroke in Finland]. PMID- 12572288 TI - [Difficulties to forecast the risk of stroke]. PMID- 12572289 TI - [Cerebrovascular stroke--the first six hours]. PMID- 12572290 TI - [Treatment of cerebrovascular stroke in a specialized stroke unit]. PMID- 12572291 TI - [What new on the way to the field of stroke prevention?]. PMID- 12572292 TI - [The results of carotis surgery in Finland]. PMID- 12572293 TI - [Rehabilitation possibilities for stroke patients in the future]. PMID- 12572294 TI - [Smoking, nicotine dependence and detoxification therapies]. PMID- 12572295 TI - [What should a medical journal's purpose be--facts or sponsored entertainment?]. PMID- 12572296 TI - [Standardized laboratory analyses for safer care. The INR work for consensus in antivitamin K treatment is a good example]. PMID- 12572297 TI - [Is placebo surgery unethical?]. PMID- 12572298 TI - [Less variation in laboratory results after the introduction of INR. Differences between hospital laboratories and laboratories within primary health care are levelled out]. AB - In 1999 a new and simplified procedure for calibration of the Owren prothrombin time (Owren PT) assay was introduced in Sweden by the national external quality assessment scheme (Equalis). The new protocol allowed local calibration by means of lyophilised national plasma calibrators and expression of results as an international normalised ratio (INR). A two-year follow-up involving analysis of data from all laboratories that have returned results to Equalis is reported. There was a significant reduction in both between-laboratory and within laboratory variation after the introduction of the new calibration procedure. For the larger hospital laboratories analysing external controls with INR > 2, the mean coefficient of variation (CV) was reduced from 9.1% to 5.6% (P < 0.0001). The corresponding results from smaller laboratories in the primary health care units showed a similar decrease in CV from 8.8% to 6.3% (P < 0.0001). This study shows that the Owren PT assay is well suited for INR calibration employing calibrant plasmas. PMID- 12572299 TI - [Rehabilitation and secondary prevention in coronary disease. Patient association's survey has yielded improved aftercare]. AB - According to national Swedish guidelines cardiovascular prevention and rehabilitation should be available for all coronary patients. Yet, over the years the access to this service has varied considerably between Swedish hospitals. Over the past five years the National Heart- and Lung Patients' Association has been engaged in a major staff training and research effort within this field assisted by a medical expert group. In assuring the quality of these efforts hospitals have been audited using a grading system of one to five hearts: one heart meaning an unsatisfactory level, five hearts an optimal level of care. An evaluation has shown that the support and grading has contributed to an improvement of preventive and rehabilitation care, though the ultimate goal of "five hearts for all" still remains to be achieved. PMID- 12572300 TI - [Coxiber and risk of myocardial infarction--the apprehension might have been exaggerated]. AB - Coxibs are a new group of non-steroidal anti-inflammatory drugs (NSAIDs), which inhibit cyclooxygenase-2 selectively. They can be prescribed for treatment of osteoarthritis, rheumatoid arthritis, primary dysmenorrhea, and acute pain. Their main advantage is a reduction in the frequency of serious gastrointestinal complications compared to NSAIDs, but they have the same tendency to induce oedema and renal side effects. The coxibs do not influence platelet function. Meta-analysis of studies with coxibs suggests that they increase the risk of heart attack, but according to recent information this may have been exaggerated. Coxibs should be used for patients at risk for gastrointestinal complications and bleeding, but perhaps not for patients at risk for arterial thrombotic diseases. PMID- 12572301 TI - [Pharmacological treatment of borderline personality disorders. Several options for symptom reduction but nothing for the disorder per se]. AB - There are few guidelines for pharmacological treatment of personality disorders. Some possible explanations for this is discussed: methodological problems, comorbidity and treatment traditions. The few controlled studies published are marred by methodological difficulties such as high frequency of drop outs, lack of compliance, short period of follow-up and differences in diagnostic procedures. The authors propose a dimensional approach focusing on cognitive, affective, anxiety and impulse dysfunction when selecting suitable pharmacological treatment. Global improvement can be seen, when target symptoms are successfully treated. Pharmacological treatment can not replace other treatment strategies such as psychosocial programs. The authors also discuss how pharmacological treatment can be understood in a psychological context. Non selective MAO-inhibitors are probably underused in Sweden. PMID- 12572302 TI - [Herbal preparations have both effects and side effects. Widespread usage dictates knowledge among physicians]. AB - The article gives a clinically oriented overview of the efficacy and safety of Ginkgo biloba, St. John's wort, ginseng, Echinacea, saw palmetto and kava based on American experiences. Clinical data support the efficacy for some of these drugs. None of them is free of adverse effects. Generally speaking, trials of herbal medicinal products have been too few, too small and too short. The lack of long-term studies is especially unfortunate since many of the drugs are used for a long time. The difference between American and Swedish legislation on herbal medicine products is described. PMID- 12572303 TI - [Still to early to tell which "speech genes" direct speech ability of human beings]. PMID- 12572304 TI - ["Doctor Death" and the mocking polyp]. PMID- 12572305 TI - [Who is to be trusted? On press reporters' assignments and agendas]. PMID- 12572306 TI - [Bad description of reality in a TV show about a general practitioner's everyday]. PMID- 12572307 TI - [Unnecessary sick listing!]. PMID- 12572308 TI - [Sanctions against Iraq hit the civil population]. PMID- 12572309 TI - [Ongoing vaccination program against cervix cancer is based on HPV test]. PMID- 12572310 TI - [Good science does not sell easily. Lakartidningen's ambition level must be adapted to already existing international demands]. PMID- 12572311 TI - [Insulin pump--25 years old and with a future. It counteracts development of late diabetic complications]. PMID- 12572312 TI - [The medical literature stands to gain from critical appraisal. Commercialism and non-contextualized results under scrutiny]. AB - Medical journal editors should put more effort into presenting scientifically correct reviews in a manner that appeals to the target audience which stands most to gain from them. In the future we need more expertly designed medical articles. All authors should present their respective conclusions. Results of controlled studies should always be presented in relation to current knowledge in the field, in order to better guide the reader in his or her daily practice. The discussion section should provide an overview of potential issues raised by the results, as well as suggestions indicating ways forward in relation to these findings. Critique and opinions emerging after publication of a study must have an impact on the way in which results are used in clinical practice. In order to reduce commercial influence, leading journals have elected to publish only those articles for which authors affirm that sponsoring companies have had no influence on the report itself. PMID- 12572313 TI - [Statistics must be seen and scrutinized! Fundamental to the progress of medical science--yet neglected in medical journals]. AB - Even the most respectable medical journals contain questionable conclusions, frequently based on erroneous statistical analyses. A common attitude among medical researchers seems to be that a deeper knowledge of statistical theory is not necessary--it might suffice to be acquainted with the statistical program packages. The most common statistical errors in medical manuscripts are related either to the structure of the data or to misunderstandings regarding the logic in some basic statistical procedures, especially significance analysis. Editors should designate professional statisticians to sort out those manuscripts which require further scrutiny from a statistical point of view. This can be organized on a consultative basis outside the framework of the traditional, "blinded" peer review system. The assessments made by the medical journals have come to play a very specific role in the Swedish academic system with respect to doctoral dissertations. Further discussion of present procedures in this area would be highly desirable. PMID- 12572314 TI - [Reporting of approval by research ethics committee and informed consent should be included in scientific information. A neglected quality indicator]. PMID- 12572315 TI - [New regulations of the Medical Society concerning professional ethics arouse suggestions to changes. That is the indication of a random sample of professionally active physicians]. PMID- 12572316 TI - [Anti-TNF-alpha treatment--an effective complement in spondyloarthropathy]. AB - Nine patients with spondylarthropathy (SpA) (6 with ankylosing spondylitis and 3 with psoriatic arthritis) who had not responded properly to conventional DMARD therapy were treated with 3 infusions (at 0, 2 and 6 weeks) of the TNF alpha inhibitor infliximab (3 mg/kg), in combination with methotrexate. To measure the effect of treatment, patients were evaluated before as well as 8 and 12 weeks after treatment with respect to disease activity (BASDAI), function (BASFI), mobility (BASMI) and global well-being in the past week (BASG1) as well as the past 6 months (BASG2). BASDAI, BASFI, BASMI and BASG1 improved significantly, with the most pronounced effect during the first 8 weeks. Also, ESR and CRP decreased significantly from baseline to 12 weeks after treatment. We conclude that infliximab is an effective therapy in SpA which does not respond sufficiently to conventional DMARD therapy. Best response to treatment was found after the first treatment regimen with regard to both effect and effect duration. Most patients responding to and tolerating anti-TNF alpha treatment needed one infliximab infusion every 8th week (3 mg/kg) to control disease activity and symptoms. Further studies are warranted to evaluate long-term outcome of anti-TNF alpha treatment in patients with SpA. PMID- 12572317 TI - [Beggars, drug abuse and care. Results from studies on begging in Copenhagen and Stockholm]. PMID- 12572318 TI - [Real physicians in the opera. Magicians, alchemists, court physicians, researchers--and a modern physician]. PMID- 12572319 TI - ["Baby walkers"--criticized already in the 18th century]. PMID- 12572320 TI - [The depressing dilemma of doctor Munthe. New reflections on a legendary physician's destiny]. PMID- 12572321 TI - [The editor-in-chief of Dagens Medicin: Josef Milerad may have become blind in one eye without knowing it]. PMID- 12572322 TI - [The debate on a new specialty subdivision delays the proposal by the National Board of Health and Welfare]. PMID- 12572323 TI - [The region Skane. Cooperation between university and health services is necessary for the development of the clinical research]. PMID- 12572324 TI - Reducing failures to communicate: a procedure-based approach to pre-surgical treatment planning. Interview. PMID- 12572325 TI - Collecting national data on hospital-acquired infections. PMID- 12572326 TI - How is health-care education to be organized in the new NHS? PMID- 12572327 TI - Dopamine agonist monotherapy in early Parkinson's disease. AB - While levodopa therapy for Parkinson's disease is still considered the gold standard, motor complications are significant disadvantages of treatment. Monotherapy with dopamine agonists may present an alternative approach with a reduced likelihood of developing dyskinesias. Further studies are required before a definitive judgment can be made. PMID- 12572328 TI - The new generation of antidepressants. AB - A major goal of antidepressant development is to improve on preceding drug classes with agents with better efficacy, tolerability and with more rapid onset of action. This article describes the new generation of antidepressants which have been recently introduced and also briefly describes possible future antidepressants which are currently being researched. PMID- 12572329 TI - Statin therapy in people with diabetes and high-risk patients. AB - Lipid lowering forms part of the treatment of atherosclerosis. Primary prevention is dependent on identification and treatment of patients at high risk of which patients with diabetes are the clearest example. This article reviews the role of lipid lowering in high-risk individuals. PMID- 12572330 TI - Safe removal of long-term cuffed Hickman-type catheters. AB - The use of long-term venous access devices is increasing. Most devices are anchored by a cuff, usually made of Dacron. The disadvantage of such cuffs is that a cut-down procedure is required for their removal. This article presents a simple technique for removal of Hickman-type catheters, emphasizing avoidance of complications. PMID- 12572331 TI - Activated protein C resistance and pregnancy loss. AB - Activated protein C resistance is a thrombophilia with an established role in producing thrombosis which more recently has been implicated in the pathogenesis of pregnancy loss. This review will analyse recent literature to evaluate this association and address the gestation and type of pregnancy loss. PMID- 12572332 TI - Neuroimaging of sexual arousal: research and clinical utility. AB - The treatment of sexual dysfunction or deviancy requires an understanding of the underlying neural substrates. Neuroimaging techniques offer insight into brain regions involved in sexual arousal and inhibition. The development of robust paradigms has implications for the assessment and treatment of sexual disorder in men and women. PMID- 12572333 TI - The use of prosthetics in hernia repair. AB - Since the early 19th century different prostheses have been tried and tested to prevent recurrences after hernia repairs. This article reviews the evolution of prostheses and their use in hernia surgery. Tension-free mesh hernia repairs using polypropylene fibre seems to be associated with very low recurrence rates and minimal complications. PMID- 12572334 TI - Percutaneous and surgical tracheostomy. AB - Percutaneous techniques for elective tracheostomy have provided a quick and relatively simple method that can be performed in the intensive care unit. Evidence-based studies comparing surgical and percutaneous tracheostomies suggest similar complication rates in trained operators of both techniques. PMID- 12572335 TI - Bladder cancer: a current update. AB - In the UK, 10,000 new cases of bladder cancer are reported per annum. Earlier diagnosis and better care have improved survival rates, but the incidence is still rising. This article updates the current understanding of bladder cancer diagnosis and management. PMID- 12572336 TI - The employment, attitudes and aspirations of non-training grade doctors. AB - Acute care in the NHS is increasingly delivered by junior doctors who receive little educational supervision. There is a continuing dramatic increase in the numbers of trust doctors. Staff grades are also increasing in number and face many frustrations. PMID- 12572337 TI - The eye in rheumatology. PMID- 12572338 TI - Prevention of the development of alcohol dependence by the early detection of alcohol misuse in accident and emergency departments. PMID- 12572339 TI - Peripartum cardiomyopathy: a forgotten diagnosis? PMID- 12572340 TI - A therapeutic dilemma: atrial fibrillation, transient ischaemic attacks and an unruptured intracranial aneurysm. PMID- 12572341 TI - Two cases of primary thyroid disease and hip fracture. PMID- 12572342 TI - Active hypothermia in the management of head injuries. PMID- 12572343 TI - Observation of a supernumerary hamstring muscle: a state of the art on its incidence and clinical relevance. AB - During routine anatomic dissection of the lower extremities of a 67-year-old male body, a supernumerary ishiocrural muscle was observed. This supernumerary muscle had similarities to a rare variant of the semimembranous muscle. On the left side it arose from the lateral dorsal side of the femur between the short head of the biceps femoris muscle and the origin of the adductor magnus muscle. It inserted on the medial condyle deep to the normal insertion of the semimembranous at the posterior aspect of the articular capsule. This muscle can be regarded either as a short deep semimembranous muscle (M. semimembranosus profondus) or as a short belly of a semitendinous biceps as known in birds. The muscle was situated closely to the vessels and nerves of the popliteal region. On the right side a similar but somewhat fainter muscle was observed whose origin emanated from the fascia of the adductor magnus muscle. The muscle probably has no major clinical importance but might be important to the surgeon who has to intervene in the popliteal region. PMID- 12572344 TI - [Standing-up/sitting-down movement. Electromyographic analysis of 4 muscles of lower limb and the erector spinae muscle: study of anticipatory postural adjustments]. AB - This study was conducted to evaluate the activity of the Erector Spinae and four other muscles of the lower limb during the 1500 first milliseconds of the sitting movement. Electromyographic activities of the Soleus, Tibialis Anterior, Biceps Femoris, Vastus Lateralis, and Erector Spinae muscles were recorded together with cinematic and kinematic data. An inhibition of the Erector Spinae activity was found at the beginning of the movement. It preceded the displacement of the first mobile kinematic marker in 8 of 15 trials. This muscular inhibition created a postural forward movement before the voluntary backward movement. This inhibition of the Erector Spinae could be considered as an anticipatory adjustment movement because of its postural function and its precocity. These results, obtained in healthy young people, should be compared with results in old people, especially with people who suffer from psychomotor dysadaptation syndrome, in whom this inhibition of the Erector Spinae seems to be absent. PMID- 12572345 TI - [Morphometric evaluation of the lateral fossa during the pre-gyrus period]. AB - During edification of neocortex, the lateral fossa is involved in the process of development of cerebral hemispheres. It changes its shape and, from a shallow depression at the end of the 3rd month, it becomes a triangular surface with marked borders. Finally, in the same time with the appearance of circumvolutions the opercles that limit it come closer and give rise to the lateral sulcus. The evolution of the lateral fossa can be analysed by linear and surface parameters. Morphometric and statistic analyse of these parameters, compared with those of the cerebral hemisphere, allowed us to establish some original criteria for appreciating the growth of the foetal brain. PMID- 12572346 TI - Three-dimensional (3-D) reconstruction of the human coccygeal body. AB - The three-dimensional reconstructions of the human coccygeal bodies were undertaken using semi-thin serial sections which were cut from the tissue specimens taken from the ventral parts of the tip of the coccyges of four patients. The coccygeal bodies were observed in the form of convoluted, irregular helical tubes. The diameters of the coccygeal bodies in serial sections were measured and a statistical analysis performed. The lumens of the coccygeal bodies were not observed in the twisted parts of the tissue specimens. Depending upon to the irregular courses of the lumens and their lack of appearances in the twisted areas, we are unable to show the three-dimensional reconstructions of the lumens of the coccygeal bodies. In conclusion, this is the first study reporting the three-dimensional reconstruction of the contours of the coccygeal body. These type of studies, which were done by using serial sections will be very helpful for the understanding of the little known organs of the human body. PMID- 12572347 TI - [Anatomical study of triple renal arteries]. AB - Within an 8 year study of the renal blood supply, the authors assessed 14 cases of triple renal arteries, 2 of them bilaterally (both on human fetuses), so a total of 16 triple renal arteries. Among the 14 cases of unilateral triple renal arteries, 8 were placed on the left side. In what concerns the caliber, the renal artery that enters the kidney through the hilum is usually greater than the others; in only three fetal samples the triple renal arteries were almost equal in caliber. Concerning the arterial traject, in 5 of the cases, two of the three renal arteries crossed one another; this aspect was assessed in first two arteries (2 cases) and last two arteries (3 cases). In 3 cases, the triple renal arteries were accompanied by double renal arteries on the opposite side. The almost parallel traject of the triple renal arteries was assessed in only 3 cases. PMID- 12572348 TI - [Variations of the celiac trunk branches in the fetus]. AB - The study was performed on 60 human foetuses, aged between 4 to 9 months, using as methods dissection and plastic and contrast substances injection. We studied the celiac trunk in what concerns the division into its terminal branches, insisting on the possible morphological variations, some rare collateral branches starting from the common arterial trunk, the dimensional relations between the branches at their origin and the level of the celiac trunk origin from the aorta, in relation with the vertebral column, the diaphragmatic passage of the aorta and with the superior mesenteric artery. We also assessed the dimensional relations (calibers at origin) between the branches of the celiac trunk. Ass possible variations of the division of the celiac trunk, we assessed: gastro-hepatic trunk, with the splenic artery directly from the aorta or from the hepatic artery; gastro-splenic trunk, with the hepatic artery originating from the aorta; hepato-splenic trunk, with origin of the left gastric artery either directly from the aorta or from the hepatic artery. Rare variations: celiaco-mesenteric trunk; two arterial trunks, hepato-splenic and hepato-gastric; separate aortic origin for all three "classic" branches of the celiac trunk; two hepatic arteries, one from the celiac trunk and the other from the aorta or superior mesenteric artery; celiac trunk that divides into several terminal branches; one or two suprarenal arteries originating from the celiac trunk. PMID- 12572349 TI - [Effects of testosterone on proliferation and IgG production of splenic lymphocytes in vitro]. AB - The purpose of the present study was to investigate the effects of testosterone on proliferative response and IgG production by splenic lymphocytes in vitro. Splenic lymphocytes from rats were cultured in the presence of the indicated doses of testosterone propionate for 72 h. The cell viability was assessed by a trypan blue exclusion test. Proliferation was determined by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and supravital stain neutral red assay. Levels of IgG and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). The proliferative response and IgG production of splenic lymphocytes in the presence of testosterone were lower than those of control cells. Testosterone may inhibit proliferative response and IgG production of splenic lymphocytes. The results also support that androgen may play an important role in immune regulations. PMID- 12572350 TI - [Effect of choroid plexus on sequestering cadmium and its pathomorphological change]. AB - To investigate whether choroid plexus can sequester cadmium(Cd) and its pathomorphological change, rabbits were given the injections of 1.0, 2.0 and 4.0 mg CdCl2/kg through ear vein, respectively. 4 and 24 hours after Cd administration, animals were sacrificed, the samples of blood, cerebrospinal fluid (CSF), choroid plexus of lateral ventricle, cerebral cortex, liver and kidney were collected and the concentration of Cd in these tissues were measured. Samples of choroid plexus were rushed in saline thoroughly, then fixed. Pathomorphological changes of choroidal epithelial cells were observed in light microscopy and electron microscopy. The result showed that concentration of Cd in choroid plexus was higher than that in blood, CSF and cerebral cortex. In high dose group, concentration of Cd in CSF was higher than that in other groups. Choroidal epithelial cells appeared vacuolation and granuler degeneration in light microscopy. Choroidal epithelial cells lost their normal slight to moderate convex luminal profiles, and appeared the loss of microvilli, shaped nuclei, increased cytoplasmic vacuolation, increased number of lysosomes, dilated rough endoplasmic reticulum, and widening of the gaps between adjacent cells. Choroid plexus could accumulate and sequester Cd to protect cerebral cortex from the fluxes of Cd in blood cadmium. When concentration of cd reached a certain level, sequestering action of choroid plexus was weak, or disappeared. Choroid plexus of lateral ventricle appeared pathomorphological change. PMID- 12572351 TI - [Study on morphology of brain tissue induced by inhale 1,2-dichloroethane]. AB - To study acute toxicity of brain tissue caused by 1,2-dichloroethane (1,2-DCE) under static inhalation. The rats were exposed to 1,2-DCE 12 hours by consecutive static inhalation. The morphology changes of brain were observed on different dose of treated rats. Dry weight and wet weight (water content) of brain were investigated too. The results showed that it is no obvious change in morphology and water content when rats were exposed to 1,2-DCE of 2.5 g/m3. The light brain edema appeared when exposed dose was 5.0 g/m3, and the average water content of brain (82.14%) was higher than control group (79.82%), which had significantly difference. The brain morphology of the 10.0 g/m3-group was significant changed compared with that of the control group. The water content of the 10.0 g/m3-group (83.65%) was significantly higher than that of the control group(79.82%) too. The study suggests that 1,2-DCE could result in brain edema when rats inhaled lots of 1,2-DCE in short time. PMID- 12572352 TI - [Quantified study on human health impact caused by coal-burning air pollution in China]. AB - To develop the mathematics model of exposure to coal-burning pollution; To confirm the exposure level of coal-burning pollutants by source analysis of atmospheric particulates; To establish the quantilification technology and methods of human health impact. Combinating the methods of epidemiology, environmental chemistry and contaminated aerography. We obtained the data of human historical expose to PM10, PM2.5, and Bap etc. that can't be obtained from the general inspect and the contribution rate of coal-burning as well as the status of coal-burning air pollutants. Confirming the degree of human health impact due to coal-burning pollutants, which included: The occurrence risk of respiratory symptoms and COPD of adults in heavily polluted area was 1.7 and 1.5 times of that of relatively clean area respectively; FVC and FEF50 of pupils decreased 194 ml and 172 ml respectively with the increasing of every unit of the Ln(PM10). FVC and FEF50 of pupils decreased 69 ml and 119 ml respectively with the increase of every unit of the Ln(SO2). Both the indices of non-specific and humoral immunity of pupils in heavily and medium polluted areas were worse than those in relatively clean area. PMID- 12572353 TI - [Estimation of arsenic accumulative intake and residents' health effects in an air pollution area--relationship between arsenic accumulative intake level and arsenicism prevalence]. AB - This paper reports the results of epidemiological survey on health effects of residents exposed to arsenic in a pollution area and estimation of arsenic accumulative intake level (EAAIL) based on calculating accumulative rice consumption and via inhalation way. 795 persons were sampled randomly from the polluted area, among whom 674 persons and 83 persons were diagnosed with Chronic Arsenic Absorption (CAA) and Chronic Arsenicism (CA) according to the National Diagnose Standard respectively. There were 60.98% CAA in 30 years old and younger age-groups and 97.59% CA in 30 years old and older age-groups. The one youngest case of CA occurred in 15 years old age group, while its EAAIL was at 1846.47 mg. The highest EAAIL was at 8706.47 mg. The rate of CA had gone obviously up at 30 years old age group. Its EAAIL was at 3833.42 mg. One equation of relationship between the rate of CA (%) and EAAIL (mg) was fitted by means of curve fitting, its is followed: Y = X1.843/e12.694 -2.866, r2 = 0.945. PMID- 12572354 TI - [Correlations between atopy and airway hyperresponsiveness]. AB - To determine the correlations between atopy and airway hyperresponsiveness(AHR), 3000 nuclear families were selected by indicator case of asthma. All of questionaire, methacholine challenge test(5 g/L) and skin test(with 12 allergens) were conducted among all relatives in those nuclear families. The data were analysed through the EP15.01 and SAS6.12 and the odds ratio(OR) was obtained. The results of the skin prick tests on 12,516 relatives of the asthma nuclear families declared that the percentages of the positive erythema and rash were 12.5% and 51.3%, respectively. Furthermore, the positive rate of male was higher than that of female(P < 0.01). At the meantime, the positive rate under age of 35 was higher than that of people with the age over 35(P < 0.01). The prevalence of AHR was 11.9% among all relatives, in which the positive erythema and rash were 15.5% and 12.7%, respectively. Both the positive erythema and rash increased the risk of AHR, and their odds ratio are 1.17 and 1.14(P < 0.01), respectively, and the risk of AHR was greater with the increasing of kind of allergens causing the positive erythema or rash. The AHR was related with positive skin prick. PMID- 12572355 TI - [Chemopreventive effect of tea and curcumin on DMBA-induced oral carcinogenesis in hamsters]. AB - 0.5% DMBA solution (in mineral oil) was applied topically to the left cheek pouch of male Syrian golden hamsters 3 times per week for 6 weeks. After the last treatment of DMBA, the animals received 0.6% green tea powder as drinking fluid, or 10 mumol curcumin applied topically 3 times per week, or the combination of green tea and curcumin treatment, or no treatment for 18 weeks. The combination of tea and curcumin significantly decreased the oral visible tumor incidence and the squamous cell carcinoma (SCC) incidence. The combination of tea and curcumin also decreased the number of visible tumors and the tumor volume as well as the numbers of SCC, dysplasic lesions, and papillomas respectively. Green tea or curcumin treatment decreased the number of visible tumor, the tumor volume and the number of SCC respectively. Green tea also decreased the number of dysplasic lesions. Curcumin also significantly decreased the SCC incidence. Tea and curcumin, singly or in combination decreased bromodeoxyuridine (BrdU)-labeling index in hyperplasia, dysplasia, and papillomas. Tea alone and in combination with curcumin significantly increased the apoptotic index in dysplasia and SCC. Curcumin alone and in combination with tea inhibited the angiogenesis in papilloma and SCC. The results suggested that green tea and curcumin had inhibitory effects against oral carcinogenesis at the post-initiation stage and such inhibition may be related to the suppression of cell proliferation, induction of apoptosis, and inhibition of angiogenesis. PMID- 12572356 TI - [Effects of tea polyphenols and tea pigments on cell cycle of HepG2 cells]. AB - The effects of tea polyphenols and tea pigments on cell cycle of hepatic cancer cells were studied. HepG2 cells were incubated with 50 and 100 mg/L tea polyphenols and tea pigments for 48 h respectively. Flow cytometry, Western blot and RT-PCR analysis were used. Flow cytometry analysis showed that tea polyphenols and tea pigments induced G1 arrest. Western blot analysis showed tea polyphenols and tea pigments significantly inhibited the expression of cyclin D1 protein and induced higher expression of P21WAFI/CIPI protein. The result of RT PCR analysis demonstrated that Cdk4 was significantly inhibited by tea polyphenols and tea pigments. It is concluded that the induction of cell cycle arrest may be an important mechanism of tea on cancer prevention. PMID- 12572357 TI - [Study on tetrazolium salt colorimetric assay for growth and survival of bacteria]. AB - To establish a tetrazolium salt(MTT) colorimetric assay for bacteria growth and survival, living or boiling killed bacteria S. aureus and E. coli were incubated with MTT in microdilution plate for 36-37 degrees C at different time. DMSO was added as formazan dissolvent and the mixtures were measured for optical density(A) or absorbence at 510 nm wave length. Living bacteria were incubated with different amount of MTT for different time. The formazan were dissolved with different amount of DMSO and the A value were measured at different wave length. The results showed that yellow MTT could be reduced into blue formazan by living bacteria S. aureus and E. coli and the A value was directly proportional to concentration of the bacteria. A values of bacteria decreased with increasing of boiling time. To some extents, absorbence of living bacteria was direct proportion to MTT concentration and incubation time in the range of 0-2 hours. DMSO was good dissolvent for bacteria formazan and the highest absorbence could be reached in wave length range of 510-530 nm. It was suggested that MTT colorimetric assay could be used for measurement of bacteria growth and survival, and its optimal parameters were as follows: MTT concentration is 0.3 mg/ml. MTT incubation time was 2 hours. DMSO amount as dissoventis was 2-fold volume of bacteria suspension. Measurement wave length was 510-530 nm. PMID- 12572358 TI - [Effects of calcium supplementation on bone mineral accretion in adolescents]. AB - The trial is conducted to evaluate the effects of calcium on bone mineral accretion in adolescents. 161 healthy adolescents aged of 9 to 20 year-old are recruited and are randomly divided into experimental groups in which 300 mg Ca is supplemented per day for each participant and control group. The dietary calcium, physical activity, SMR, distal one-third radial bone mineral content (BMC), bone width (BW), height, weight and serum total ALP are measured at both the beginning and the end of the trial. It is found that the average calcium intake from dietary sources is 500 mg/d in both groups. Increasing daily calcium intake via calcium supplementation in adolescents results in a moderate increases of BMC, and there is a significant increase of BMC in the females during the late adolescence. No significant differences in anthropometry are observed between the two groups at the end of the study while the increase of serum ALP level in the females in experiment group is significantly lower than that in the control group during the late adolescence. It is concluded that during the pubertal years when the BMC accretion is evident, there are no significant differences in bone mass accrual between the two groups. While in the females, calcium supplementation may help enhance the bone mineral accrual during late adolescence when the BMC accretion is remarkably reduced. PMID- 12572359 TI - [Effect of isoflavone on human breast cancer cell line MCF-7 and its probable mechanism]. AB - The effects of isoflavone on human breast cancer cell line MCF-7 growth and apoptosis and its probable mechanisms were studied in vitro. The results showed that isoflavone could significantly inhibit the growth and induce the apoptosis of MCF-7 cells. Western Blotting showed that isoflavone increased iNOS protein expression. These results suggested that isoflavone significantly inhibited the MCF-7 growth and induced cell apoptosis by regulating iNOS gene expression. PMID- 12572360 TI - [Influences of diet and nutrition on obesity of pre-school children]. AB - OBJECTIVE: To study diet and nutrition on obesity of pre-school children. METHODS: Applying food weighing and food record method, to understand the diet situation of toddler at kindergarten and plus home in a week. RESULTS: The energy intake in obese children was 1557.0 kcal that was higher than 1442.4 kcal in the control group (P < 0.001). Calculating the intake of nutrients, we found that protein, fat, and CHO in obese children were higher than in the control group. Significant difference in fat and CHO was detected between the two groups. CONCLUSION: Dietary fat intake is a risk factor for childhood obesity. To prevent childhood obesity, parents need to help children to develop good food habit. PMID- 12572361 TI - [Association between the excretion levels of urinary TNOC and NAAs in the inhabitants of high and low risk area for esophageal cancer in southern China]. AB - To assess the role of urinary total N-nitroso compounds(TNOC) index in study etiology of N-nitroso compounds(NOC) of esophageal cancer in southern China and to study the association between TNOC index and traditional indexes of N nitrosamino acids(NAAs). The samples of 12-hr overnight urine were collected from 120 male adults in high-risk area (Nanao county) and low-risk area(Lufeng county) for esophageal cancer. The 240 male healthy subjects(35-64 years old) were selected by a 3-stage random cluster sample procedure. Levels of TNOC and 4 categories of NAAs were measured in samples. The results showed that Urinary TNOC contents were positively correlated with esophageal cancer mortality in the south(rs = 0.25, P < 0.05). Only 7.5% content of urinary TNOC came from the 4 categories of NAAs in the high-risk area when 57% content of urinary TNOC came from the NAAs in the low-risk area. The urinary NOC components in the high-risk area were more complex than the components in the low-risk area. Urinary TNOC is a better index that is applied to assess the exposure dose of TNOC in humans and study etiology of NOC of esophageal cancer. When the categories of NOC in urine are less, TNOC index is correlated with NAAs indexes. The results imply that TNOC are implicated in the etiology of esophageal cancer in southern China. PMID- 12572362 TI - [Dietary status of preschool children from day-care kindergartens in six cites of China]. AB - In order to highlight nutrients of potential concern on deficiency for the age groups under study, the dietary status of preschool children were studied in the kindergartens of Beijing, Shanghai, Guangzhou, Chengdu, Changsha and Dalian in 1998-1999. All the children who regularly attended the kindergarten, who were in the age range of 3-6 year old and generally health were considered eligible for enrollment in this study. The final results included a total of 1170 children, with 583 boys and 587 girls. Food weighing method was used in consecutive three day dietary survey by recording breakfast, lunch and afternoon refreshments in kindergarten. Questionnaire form was applied to record the other food consumption outside of the kindergarten. Nutrient intakes of per child were calculated according to the Chinese Food composition Table. The average energy, protein, iron, selenium, vitamin A, vitamin E, thiamin, riboflavin, and nicotinic acid intakes were adequate in each group. The dietary energy provided by fat was near to the high marginal (30%), which indicated that more fat intake should be avoided in these children. The ratio of energy provided by each meal per day was lower in the breakfast and higher in the dinner and foods consumed at home. The average ratio of calcium to phosphorus for 3-6 years was 0.63. A deficiency of calcium is rather common, and the intake of calcium accounted for only 61.6% of the recommended nutrient intake(RNI). Salt intake was relatively higher than that of adequate intake recommended by Chinese Nutritional Society. The zinc intake reached 62.9% of RNIs. Vitamin C intakes from each age of groups did not meet their RNI. The present study indicates that the deficiencies of some trace nutrients in the diets for preschool children in day-care kindergartens is probably related to that the body weight and height of preschool children have not achieved a "satisfactory" level. PMID- 12572363 TI - [Effect of coal-burning air pollution on children immune function]. AB - Study of effect of coal-burning air pollution on children immunity function was hold in Taiyuan city which is one of typical coal-burning air pollution city in China. The children of grade 4-5 who dwell more than 3 years were screened in three different pollution areas. The test includes the observation on the T lymphocyte subpopulations classes (CD3+, CD4+ and CD8+), saliva lysozyme contents and immunoglobulin content. The results showed that the contents of saliva lysozyme and immunoglobulin were lower than those in control area respectively(P < 0.05), and T lymphocyte subpopulations class (the percent ages of CD3+, CD4+ and CD8+) had the same trend, but it was not marked. The results suggested that coal-burning air pollution could affect children by nonspecific immunity and sap immunity. The effect on cell immunity was not marked. PMID- 12572364 TI - [Study on dynamic adsorption of HF gas by ion-exchange fiber]. AB - The dynamic adsorption capacity of HF gas utilizing domestic FA-weak basic ion exchange fiber as adsorbent is studied. Also, the factors influencing dynamic adsorption are investigated and the re-adsorption effect of HF gas is tested. The results showed that the pierce through adsorption amount of HF by ion-exchange fiber is 75 mg/g, and the saturated adsorption amount of HF by the fiber is 152 mg/g. There is no obvious change in adsorption effect after a repeated 5-run regeneration of the fiber, showing its good chemical stability. PMID- 12572365 TI - [Photocatalytic oxidation of airborne VOCs on TiO2]. AB - Titanium dioxide(TiO2) was used to catalyze the photolysis of benzene, xylene, n heptane, methanol, acetone, ethylether, formaldehyde, trichloroethylene and perchloroethlene in air. The photolysis was carried out with a high pressure mercury lamp. The degradation rate of these VOCs was more than 80 percent in five minuts except formaldehyde. The effects of catalyst amount, different types of TiO2 catalyst and density of UV on the degradation rate of the VOCs were also studied. PMID- 12572366 TI - [Application of an ozone disinfection apparatus for small drinking water supply systems in rural areas]. AB - To design an ozone disinfection apparatus which is inexpensive and to practical for small drinking water supply system in rural areas. And test its effectiveness. Designed apparatus must be rapid, low cost, high automatic and easy for operation. The key technologies are to create ozone of high concentration and to mix it with water very well, and add it into main water pipe automatically. The results showed: In laboratory, at the condition of ozone of 0.2 mg/l for 3 minutes, all germs and bacteria coli were killed. In the field study, the water samples from three villages met the standard for drinking water very well. THE CONCLUSIONS: The apparatus reached the design requirement. An innovative part of the study is that we have improved a device for mixture of ozone with water. The laboratory and field study have proved the apparatus is low cost, high effectiveness, and easy to operate u. It is very useful for small drinking water supply system in rural areas. PMID- 12572367 TI - [Method for multiple gaseous pollutants monitoring by passive sampler]. AB - A new monitoring method used passive samplers by molecular diffusion was developed to determine simultaneously the concentration of HCHO, NO2 and SO2 in the air. Pretreated activated-charcoal fibers were used as absorbent layer. After being sampled, the gaseous pollutants collected in absorbent layer were desorbed by 0.05 mol/lNaOH solution and oxidized by 0.3% H2O2 to HCOO-, NO2- and SO4(2-), finally were analyzed using ion chromatographic techniques. The passive sampler can be used in the condition of face velocity of 0.2-1.5 m/s, temperature of -10 35 degrees C and relative humidity of 20%-80%. The sample rates for HCHO, NO2 and SO2 are 67 ml/min, 80 ml/min and 64 ml/min, respectively. The relative standard deviations are 7.3%, 3.6% and 6.8%. The lowest quantitative detection concentrations of 24 h sampling for HCHO, NO2 and SO2 are 0.026 mg/m3, 0.008 mg/m3 and 0.023 mg/m3 respectively. In field comparative test with active sample method, the new method show that its relative overall uncertainty for HCHO, NO2 and SO2 are 23%, 23% and 18%. By the method, sampling and analyzing three kinds of gaseous pollutants simultaneously can be realized, and the foreground of application of diffusive sampling technology can be expanded. PMID- 12572368 TI - [Effects of tea polyphenols on mice survival rate and white blood cell count after radiation]. AB - In order to evaluate the anti-radiation effect of TPP (tea polyphenols), 60 female Kunming mice were chosen as group I for WBC (white blood cell) count, and another 60 female Kunming mice were set as group II for survival test. TPP was administrated orally to the mice. The results revealed that no significant difference was observed in the number of white blood cells in every test after the irradiation compared with that of the control group. Consequently, TPP showed no observably effect on the protection of the number of WBC against high dose irradiation. However, Mice survival rate of 0.83 ml/kg BW group and 25.0 ml/kg BW group are remarkably higher than that of the control group(P < 0.01). On the contrary, the control group had no survivor 30 days after the irradiation. This suggested that TPP can increase the survival rate evidently on mice after irradiation. PMID- 12572369 TI - [Recent advances for research on chemopreventive effect of beta-carotene]. AB - Cancer is the culmination of the chronic disease process, and can be attributed the interactions of environment and genes. An optimistic message for 35% cancer prevention may be in dietary constituents, food supply. Carotenoids are isoprenoid compounds synthesized in plants and microorganisms, but not in animals. Human tissue contains only a fraction of the total number of carotenoids(nearly 600 have been identified in nature) present in food supply such as fruit and vegetables. People with high intake of beta-Carotene have a reduced risk of cancer. This outcome seems to bear relation to the chemical to quench singlet oxygen and inhibit peroxyl free radical reaction. But the ATBC and CARET cancer prevention study found the increase in lung cancer incidence in smokers and asbestos. This adverse effect may be explained by beta-Carotene prooxidant when tobacco smoke exposure and oxidative stress occurs, and cellular DNA damage was further aggrevated. Phase I enzymes were induced. Cell transformation was enhanced. PMID- 12572370 TI - [Hazards of radio frequency magnetic field and their prevention and control]. AB - The paper briefs the sources of radio frequency (RF) electromagnetic field, their hazards to human health and the hygienic standards of RF electro-magnetic field for occupational exposure and for general population exposure. Based on the physical properties of RF electromagnetic field, the author put forward the comprehensive measures for controlling the electromagnetic radiation to meet the hygienic standards. These measures are improvement of the technology to lower the intensity of RF electromagnetic radiation, shielding the RF radiation sources to reduce the radio frequency levels in accessible areas, increasing the distance between the sources and human, and using personal RF protecting devices in area of high radio frequency levels, etc.. In protecting the general population from exposing to RF radiation, measures keeping a safe distance between the sources and the public play very important role, in which good city planning and the effective control of new and existing RF sources are very important. Referring to the RF electromagnetic problems by using electrical devices in the household, different measures based on the above principles for controlling RF hazards are also cited. PMID- 12572372 TI - [Modernization of Chinese Materia Medica and I]. PMID- 12572371 TI - [Diagnosis and treatment of rheumatoid arthritis and ankylosing spondylitis]. PMID- 12572373 TI - [Clinical observation on treatment of rheumatoid arthritis by combined therapy with methotrexate, sulfasalazine and Chinese herbal medicine]. AB - OBJECTIVE: To observe the effect of Fengshi No. 1 (FS1) in treating patients with active stage of rheumatoid arthritis (RA). METHODS: Patients with RA were randomly divided into two groups, the 40 patients in the treated group were treated with combined therapy of methotrexate (MTX), sulfasalazine (SSZ) and FS1, and the 20 in the control groups were treated with MTX and SSZ alone. RESULTS: In the treated group, the total effective rate was 97.5%, the clinical controlled and markedly effective rate 95.0% and the occurrence rate of side-toxic reaction 10.0%, as compared with those in the control group, 60.0%, 20.0% and 45.0% respectively, the difference was significant (chi 2 = 11.91, 32.23 and 7.67 respectively, all P < 0.01). The effect in the treated group was superior to that in the control group in abating joint swelling and pain, improving function of joint, reducing immune indices and ameliorating iconographic features (P < 0.01 or P < 0.05). CONCLUSION: FS1 not only has the effects of anti-inflammation, analgesis, regulating immune reaction, but also could retard the occurring of bone destruction, reduce the toxic-side effects of MTX and SSZ. PMID- 12572374 TI - [Clinical observation on treatment of ankylosing spondylitis with huatan zhuyu jiedu decoction]. AB - OBJECTIVE: To observe the clinical effect of Huatan Zhuyu Jiedu Decoction (HZJD) in treating ankylosing spondylitis (AS). METHODS: One hundred AS patients were randomly divided into two groups, the 60 patients in the treated group were treated with HZJD, the 40 patients in the control group were treated with Duhuo Jisheng Decoction. Treatment for both groups was orally given 1 dose per day in decoction form for 1 month as a treatment course. The efficacy of treatment was evaluated after 1-3 treatment courses. RESULTS: The effect of treatment in the treated group was short-term controlled in 12 patients, markedly effective in 29, improved in 16 and ineffective in 3, the total effective rate was 95% (57 cases), while in the control group, the corresponding figure was 2, 8, 20, 10 and 75% (30 cases), comparison between the two groups showed significant difference (chi 2 = 20.85, P < 0.01). The manifestation of symptoms of patients in both groups improved obviously, the pain in loin and back alleviated, spinal motion (flexion forward, extension backward and lateral inclination) improved and thoracic expansion increased. Laboratory indices such as ESR, C3, IgA and CRP reduced obviously after treatment (P < 0.01 or P < 0.05), and better effect was shown in the treated group (P < 0.05). X-ray film after treatment showed improvement in 11 cases in the treated group and 4 in the control group, inter-group comparison showed significant difference (chi 2 = 7.99, P < 0.01), the effect of HZJD was better than that of the control. CONCLUSION: HZJD has definite therapeutic effect in treating AS. PMID- 12572375 TI - [Clinical randomized double-blinded study on treatment of vascular dementia by jiannao yizhi granule]. AB - OBJECTIVE: To observe the effect of Chinese herbal medicine in treating vascular dementia (VD) and to explore the method for therapeutic effect evaluation. METHODS: Multi-center, double-blinded randomize-controlled method was adopted to observe the effect of treatment of 242 patients with VD of mild or middle degree, they were 89 cases in the TCM group, 106 cases in the western medicine group and 47 in the placebo group. Minimental state examination (MMSE), Blessed dementia scoring on TCM symptom classified evaluation were used to evaluate the therapeutic effect after 60 days treatment. RESULTS: The total effective rate of Jiannao Yizhi Granule was 58.4%, showing a better effect than that of the placebo group and the western medicine group respectively. The treatment was superior in holistic regulation and systemic functional state improvement, it could not only improve the mild middle key symptoms of VD and the mental disorder induced reduction of functional activities, but also alleviate or eliminate the peripheral symptoms of VD. CONCLUSION: The effect of treatment of vascular dementia by Jiannao Yizhi Granule is certain and is worth spreading. PMID- 12572376 TI - [Clinical and experimental study on treatment of migraine with shutianning granule]. AB - OBJECTIVE: To explore the cause and pathogenesis of migraine based on TCM theory and modern researches to seek for new thinking and effective drugs in treating migraine. METHODS: Ninety patients with migraine were divided into three groups, the treated group treated with Shutianing Granule (STNG) 9 g, three times a day, the control A group treated with composite Yangjiao capsule, 5 capsules each time, three times a day and the control B group treated with flunarizine hydrochloride capsule, 5 mg, once a day. The treatment lasted for 28 days to observe the degree, lasting time and frequency of pain attack, and the headache index was calculated. Transcranial Doppler examination, serum beta evoked potential (beta-EP) and neuropeptide (NPY) determination were also performed before and after treatment. In the experimental study, effect of treatment of cerebral NPY and beta-EP were tested in SD rats with chronic pain in the treated or the control groups. RESULTS: In the treated group, the markedly effective rate was 56.67% and the total effective rate was 90.00%, significant difference was shown in comparison with those in the control B group (P < 0.05), but with no difference in comparison with those in the control A group. Cerebral beta-EP level was raised and plasma NPY level was lowered in all the three treated groups. Compared with the normal saline group, there was significant difference (P < 0.05 or P < 0.01). CONCLUSION: STNG could improve the cerebrovascular function effectively, raise plasma beta-EP level, lower plasma NPY level and alleviate vascular tension in patients with migraine. PMID- 12572377 TI - [Clinical observation on effect of yishen jiedu recipe in retarding the course of chronic renal failure]. AB - OBJECTIVE: To observe the effect of Yishen Jiedu Recipe (YSJDR) in retarding the course of chronic renal failure (CRF). METHODS: Forty-five patients were divided into two groups, the 22 patients in Group A were treated with low-protein diet and blood pressure controlling therapy only, while the 23 patients in Group B were treated with additional YSJDR. The time (month) for doubling serum creatinine (SCr) level was taken to evaluate the speed of CRF progression. RESULTS: In Group A, the time for SCr increased from 288.4 +/- 96.7 mumol/L to 586.3 +/- 251.3 mumol/L was 16.7 +/- 5.1 months, while in Group B the time for SCr increased from 291.2 +/- 101.2 mumol/L to 589.6 +/- 257.5 mumol/L was 28.2 +/ 8.7 months. Comparison between the two groups showed significant difference (P < 0.05). CONCLUSION: On the basis of low-protein diet and blood pressure controlling therapy, the additional treatment of YSJDR could markedly retard the progression speed of CRF. PMID- 12572378 TI - [Effect of jianpi wenshen decoction on serum gastrin, plasma motilin and somatostatin in patients of diabetic diarrhea]. AB - OBJECTIVE: To observe the effect of Jianpi Wenshen Decoction (JWD) on serum gastrin, plasma motilin and somatostatin in patients of diabetic diarrhea (DD). METHODS: Patients with DD were randomly divided into two groups, the JWD group and the control group treated with Loperamide (LPA). Besides, a normal control group was set up. Changes of serum gastrin, plasma motilin and somatostatin were observed. RESULTS: Before treatment, the levels of gastrin and motilin in both groups were higher and somatostatin lower than those in the normal control group. After 1 month treatment, levels of the three indices were restored in both group approaching the normal range with insignificance as compared with those in the normal control group (P > 0.05). Level of plasma motilin and serum gastrin showed an increasing trend along with the therapeutic effect elevation, while level of somatostatin showed a decreasing trend. CONCLUSION: JWD could promote the recovery of the impaired function of vegetative nerve system in DD patients. At the same time, serum gastrin, plasma motilin and somatostatin may be taken as the indexes for evaluating the efficacy in treating DD. PMID- 12572379 TI - [Clinical and experimental study on effect of jietongyin lotion in treating vulvovaginitis in young girls]. AB - OBJECTIVE: To study the clinical effect and mechanism of Jietongyin Lotion (JTYL) in treating vulvovaginitis in young girls. METHODS: Patients were randomized into the JTYL group (n = 300) and the control group (n = 60, treated with Rishu an Lotion). Comparative observation on scoring of main symptoms and signs in them was carried out, and the pharmacological and pharmacodynamic experiment was also conducted on effect of JTYL in animals. RESULTS: In the JTYL group, the total effective rate was 98% and was 93% in the control group. The cure rate in the two groups was 93% and 83% respectively, which was significantly higher in the JTYL group (P < 0.05). Experimental study showed that JTYL has effects in anti inflammation, bacteriocidal, itching relieving, pain releasing, and immune regulation. It showed non-irritatory on mucous membrane. CONCLUSION: JTYL has good reliable effect in treating vulvovaginitis in young girls without any obvious adverse effects, showing anti-inflammatory, bacteriocidal, itching relieving, pain releasing and immune regulation. PMID- 12572380 TI - [Effect of Chinese drugs mixture on immune function of patients with extremely severe burn]. AB - OBJECTIVE: To study the regulatory effect of Qinghuo Baidu Yin (QHBDY, a mixture prepared with Chinese drugs) on immune function of patients with extremely severe burn (ESB). METHODS: Thirty patients with ESB were divided into two groups, conventional therapy was given to both groups, but QHBDY was given to the treated group additionally. Immunological indices, including peripheral blood T lymphocyte subsets, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) were determined 3 weeks after treatment to evaluate and compare the therapeutical effect in the two groups. RESULTS: Compared with the control group, CD3, CD4, CD4/CD8, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) levels were markedly decreased in degree, and recovered earlier and quicker, with CD8 increased mildly (P < 0.01) and turned back more quickly. And so did the parameters of the treated group in comparing with that of the control group at anytime (P < 0.05 or P < 0.01). CONCLUSION: Chinese drugs mixture shows the regulatory effect on both cellular and humoral immune function in patients with ESB. PMID- 12572381 TI - [Clinical observation on effect of kesuning granule in treating acute onset of chronic bronchitis]. AB - OBJECTIVE: To prove the effect of Kesuning Granule (KSNG) in treating chronic bronchitis in exacerbation (Phlegm-Heat syndrome) and objectively evaluate its safety. METHODS: The double-blinded, double-dummy and randomized controlled method was adopted to observe 120 patients, who were divided into the treated group (n = 60, treated with KSNG 8 g, three times a day) and the control group (n = 60, treated with Jinbei Tankeqing Granule (JBTKQ) 7 g, three times a day). The therapeutic course for both groups was 6 days. RESULTS: In the treated group, the markedly effective rate was 58.33% and effective rate was 93.33%, while in the control group, they were 51.67% and 91.67% respectively, no significant difference was found between the two groups. For therapeutic effects on TCM syndromes, the markedly effective rate and effective rate in the treated group was 68.33% and 93.33% respectively and in the control group, 66.67% and 90.00% respectively, also showed insignificant difference. The effect initiative time was earlier in the treated group than that in the control group significantly (P < 0.05). No adverse effect was found in the observation. CONCLUSION: KSNG shows a definite clinical effect with no obvious toxic-adverse effects. PMID- 12572382 TI - [Therapeutic effect of xinfeng capsule in treating adjuvant arthritis in rats and its effect on fas, fasL and bcl-2 expression in synovial membrane]. AB - OBJECTIVE: To observe the therapeutic effect of Xinfeng Capsule (XFC) in treating adjuvant arthritis of rats and its effect on fas, fasL and bcl-2 expression in synovial membrane. METHODS: Rats were randomly divided into the normal group, the model group, the model group, the methotrexate (MTX) group, the TPT group and the XFC group. Except for the rats in the normal group, animals were modelled to adjuvant arthritis with Freund's complete adjuvant, and the latter three groups were treated with MTX, Tripterygium wilfordii polysaccharide (TPT) and XFC respectively. The arthritis index (AI), change of body weight (BW) of rats were recorded, and the expressions of fas, fasL and bcl-2 in rats' synovial membrane were determined. RESULTS: Compared with before treatment, the AI in the three treated groups was lowered significantly (P < 0.01, P < 0.05). The BW increment in the XFC group after treatment was insignificantly different to that in the normal group (P > 0.05), while it was significantly lower in the other two treated groups than that in the normal group and the XFC group (P < 0.01). Compared with the model group, fasL expression was increased in the XFC group significantly (P < 0.05), but bcl-2 expression decreased, fas expression showed insignificant difference (P > 0.05). Comparison of the three gene expressions between the three treated groups showed no significant difference (P > 0.05). CONCLUSION: XFC could decrease AI in rats with adjuvant arthritis in the same way as MTX and TPT, it effected better in increasing BW than the latter two. The effect of XFC might be performed by its action in enhancing fasL expression, inhibiting bcl-2 expression and promoting apoptosis of proliferated synovial membranous cell so as to restrain hyperplasia of synovial membrane. PMID- 12572383 TI - [Effect of tiaoxin recipe on learning and memory related gene expression in hippocampus of senescence accelerated mice]. AB - OBJECTIVE: To study the effect of Tiaoxin Recipe (TXR) on learning and memory related gene expression in hippocampus of senescence accelerated mice (SAM). METHODS: Changes of learning and memory related gene expression, including mineralocorticoid receptor (MR), presenile protein 1 and 2 (PS-1, PS-2), tau, APP, apoE and bcl-2 in hippocampus of SAM were determined by reverse transcription polymerase chain reaction (RT-PCR). The effect of TXR were tested. E2020 was used as the drug for control. RESULTS: Compared with those in the same aged mice, in the 5-month old SAM, levels of gene expression of MR, tau, PS-2 and APP were significantly higher, that of apo-E lower, levels of gene expression PS 1 and bcl-2 were unobviously changed; while in the 12-month old SAM, gene expression of MR and tau were higher, bcl-2 was lower and PS-1, PS-2, apoE and APP were also unobviously changed. Continuously orally taken TXR could correct the abnormality of MR, tau and apoE gene expression in hippocampus of 5-month SAM and that of MR and bcl-2 in 12-month SAM. CONCLUSION: Continuously orally taken of TXR has the effect of regulating and correcting learning and memory related gene expression in hippocampus of 5-month and 12-month SAM. PMID- 12572384 TI - [Effect of Salvia miltiorrhiza on neuropeptide Y1-36 and calcitonin gene-related peptide in neonatal rats with hypoxia-ischemic brain injury]. AB - OBJECTIVE: To observe the effects of Salvia miltiorrhiza (SM) on levels of neuropeptide Y1-36 and calcitonin gene-related peptide immune reactive substances (ir-NPY, ir-CGRP) in blood plasma and pons-oblongata after hypoxia-ischemic brain injury (HIBI) in neonatal rats. METHODS: Seven-day old rats were randomized into HIBI group (A), HIBI + SM group (B) and sham operation group(C). And each group was subdivided into 4 subgroups according to the different time after operation. 0.5 ml SM was injected intraperitoneally immediately and every 12 hrs afterwards. Changes of ir-NPY and ir-CGRP levels in plasma and pons-oblongata were observed immediately and 12, 24 and 48 hrs after HIBI by radioimmunoassay. RESULTS: Plasma levels of ir-NPY and ir-CGRP in different times after HIBI were all significantly raised but those in pons-oblongata were either raised or lowered to a certain degree. Part of the elevated ir-NPY could be reversed by SM injection. CONCLUSION: Central and peripheral neuropeptide Y1-36 and calcitonin gene-related peptide take part in the pathophysiological process of HIBI, SM could partially reverse the abnormal post-HIBI elevation of ir-NPY, which may be one of the pathways of SM in promoting recovery of damaged brain function. PMID- 12572385 TI - [Experimental study on effect of sini decoction on myocardial endothelin in myocardial ischemic rats]. AB - OBJECTIVE: To study the molecular mechanism of effect of Sini Decoction (SND) on myocardial endothelin (MET) in myocardial ischemic rats. METHODS: Sprague-Dawley rats were randomly divided into 3 groups, the normal control group, the ischemia group and the SND group. Myocardial ischemia was produced by pituitrin in the latter two groups. The content, immunohistochemical assay and gene expression of MET-1 were determined in all the three groups and compared. RESULTS: The content of MET in the SND group was significantly lower than that in the ischemia group (P < 0.01). Immunohistochemical examination showed that MET-1 was mainly located at the cardiac muscle cells and vascular endothelial cells with the grey scale obviously lower in the ischemia group than that in the control group and the SND group (P < 0.01). While RT-PCR showed that the grey scale of PCR product band in the ischemia group was significantly higher than that in the other two groups (P < 0.01). CONCLUSION: SND could significantly lower MET content, it may be related to the effect of SND in inhibiting MET-1 gene expression and protein synthesis. PMID- 12572386 TI - [Effect of astragalus-angelica mixture on osteopontin expression in rats with chronic nephrosclerosis]. AB - OBJECTIVE: To observe the effect of Astragalus-Angelica Mixture (AAM) on osteopontin (OPN) expression in rats with chronic nephrosclerosis. METHODS: Chronic nephrosclerosis model rats induced by repeated intraperitoneal injection of puromycin were randomly divided into the model group, AAM group and Irbesartan (an antagonist of angiotensin) group. The experimental course lasted 12 weeks. Blood and urine samples were examined by biochemical method. Kidney tissue was taken for pathological stain and immunohistochemical method and was applied to examine OPN expression, mononuclear macrophage, laminin in extracellular matrix and decorin expressions. RESULTS: AAM showed the effects of decreasing urinary protein and improving renal function similar to that of Irbesartan. It also could alleviate the pathological damage of kidney tissue, especially in decreasing renal tubular mesenchymal damage index. The accumulation of decorin and laminin in the mesenchymal extracellular matrix significantly decreased. Renal tubular OPN expression and mesenchymal infiltration of mononuclear macrophage decreased significantly and in a positive correlated manner (r = 0.885, P < 0.01). CONCLUSION: AAM has similar renal protective action to that of Irbesartan, this action may be related to the inhibition of up-regulated OPN expression. PMID- 12572387 TI - [Effect of kanglaite injection combined with Chinese drug therapy according to syndrome differentiation on quality of life and immune function in patients with advanced lung cancer]. PMID- 12572388 TI - [Clinical observation on auxiliary effect of xiaochuan paste in preventing repeated respiratory infection in child]. PMID- 12572389 TI - [Clinical observation on effect of qirong runchang oral liquid in treating constipation after stroke]. PMID- 12572390 TI - [Study on relationship of endothelin and blood stasis syndrome]. PMID- 12572391 TI - [Analysis of English translation layers of often used terms of Traditional Chinese medicine]. PMID- 12572392 TI - [Survey of skin adverse reaction induced by Chinese herbal medicine]. PMID- 12572395 TI - Finding time. PMID- 12572393 TI - [Progression of study on promoting ovulation by Chinese herbal medicine]. PMID- 12572396 TI - NHS University--a better future? PMID- 12572397 TI - Elective placement. United Hospitals Network, Toronto. PMID- 12572398 TI - Sir Douglas Black. Obituary. PMID- 12572400 TI - Psychosurgery. AB - The use of surgical procedures to alter mental states raises many issues. Surgery on the brain has been known for thousands of years, but procedures developed in the 1930s, '40s and '50s, and the reasons for them, raised many ethical issues that remain with us today. The following article touches on the history of psychosurgery, the conditions treated, the literature on the subject, and the ethical and legal issues. PMID- 12572401 TI - Oral and maxillofacial surgery in Tennessee. PMID- 12572402 TI - The Dr. Jack E. Wells Memorial Dedication to Dentistry Award goes to Dr. R. Benjamin Alley. PMID- 12572403 TI - Self inflicted shotgun wound to the face: a case report. AB - Injuries caused by firearms often test the ingenuity of the surgical team. The variables encountered related to the hard and soft tissue destruction make each case unique. The only way to achieve predictable outcomes with these patients is to treat a large enough number that you are guided by your previous experience. This patient was another opportunity to add to our knowledge base for treating this difficult class of patient. PMID- 12572404 TI - Cleft craniofacial. PMID- 12572405 TI - The oral and maxillofacial surgeon's role in the diagnosis and treatment of oral cancer. PMID- 12572406 TI - Management of complex multi-space odontogenic infections. AB - The successful management of multi-space orofacial odontogenic infections involves identification of the source of the infection, the anatomical spaces encountered, the predominant microorganisms that are found during the various stages of odontogenic fascial space infection, the impact of the infectious process on defense systems, the ability to use and interpret laboratory data and imaging studies, and a thorough understanding of contemporary antibiotic and supportive care. The therapeutic goals, when managing multi-space odontogenic infections, are to restore form and/or function while limiting patient disability and preventing recurrence. Odontogenic infections are commonly the result of pericoronitis, carious teeth with pulpal exposure, periodontitis, or complications of dental procedures. The second and third molars are frequently the etiology of these multi-space odontogenic infections. Of the two teeth, the third molar is the more frequent source of infection. Diagnostic imaging modalities are selected based on the patient's history, clinical presentation, physical findings and laboratory results. Periapical and panoramic x-rays are reliable initial screening instruments used in determining etiology. Magnetic resonance imaging and computed tomography are ideal imaging studies that permit assessment of the soft tissue involvement to include determining fluid collections, distinguishing abscess from cellulitis, and offering insight as to airway patency. Antibiotics are administered to assist the host immune system's effort to control and eliminate invading microorganisms. Early infections, first three (3) days of symptoms, are primarily caused by aerobic streptococci which are sensitive to penicillin. Amoxicillin is classified as an extended spectrum penicillin. The addition of clavulanic acid to amoxicillin (Augmentin) increases the spectrum to staphylococcus and other anaerobes by conferring beta-lactamase resistance. In late infections, more than three (3) days of symptoms, the predominant microorganisms are anaerobes, predominantly Peptostreptococcus, Fusobacterium, or Bacteroides, that are resistant to penicillin. Clindamycin is an attractive alternative drug for first line therapy in the treatment of these infections. The addition of metronidazole to penicillin is also an excellent treatment choice. Alternatively, Unasyn (Ampicillin/Sublactam), should be considered. The mainstay of management of these infections remains appropriate culture for bacterial identification, timely and aggressive incision and drainage, and removal of the etiology. It is usually preferable to drain multi space infections involving the submandibular, submental, masseteric, pterygomandibular, temporal, and/or lateral pharyngeal masticator spaces, as early as possible from an extraoral approach. Trismus and airway management are important considerations and may preclude the selection of other surgical approaches. The patients with multi-space infections should be hospitalized and patient care provided by experienced clinicians capable of management of airway problems, in administration of parenteral antibiotics and fluids, utilization of interpretation of laboratory and diagnostic imaging studies, and control of possible surgical complications. PMID- 12572407 TI - Obstructive sleep apnea: a case report. PMID- 12572408 TI - Combined orthognathic and facial aesthetic surgery with case reports. AB - There are multiple hard- and soft-tissue considerations in facial surgery. The expertise of the oral and maxillofacial surgeon in treating patients with multiple facial injuries has shown time and time again that hard- and soft-tissue reconstruction can be performed either as simultaneous or as delayed procedures. The benefits of a total facial analysis approach to facial surgery are based upon aesthetic, functional, rejuvenative, and self-esteem issues. Proper diagnosis coupled with a comprehensive treatment plan utilizing a team approach provides for both simultaneous and delayed procedures to be completed and can provide for safer and more predictable outcomes. The expanding range of possibilities exists to provide patients with treatment options for improvement in the aesthetic, functional, and rejuvenative aspects of their facial features. A rather fast metamorphosis has taken place from the late 1960s to the present period. This has carried orthognathic surgery from its infantile state of a technical procedure to a sophisticated surgical procedure(s) creating an outcome that is unsurpassed by any other specialty. PMID- 12572409 TI - Guidelines for management of disorders of the temporomandibular joint and related structures. PMID- 12572410 TI - Maxillofacial treatment by dentists in the U.S. military. PMID- 12572412 TI - Associated structures. PMID- 12572411 TI - Management of complex facial fractures. PMID- 12572413 TI - [Accumulative trends of ephedrine and pesudoephedrine in Ephedra sinica]. AB - In Yi Kezhao Alliance, the content of ephedrine and pseudoephedrine in Ephedra sinica is in a ratio of four to six. The total content of ephedine and pseudoephedrine form cultivated Ephedra sinica is higher than that of wild one, and in direct ratio with the years of plant. For the content enhance of total alkaloids, decreasing irrigation and applying middlingly fertilizer is more effective measure. PMID- 12572414 TI - [Pharmacognostical identification on fruits of Actinidia species (II)]. AB - The observation of powder were carried out on fruits of 10 kinds of plants belonging to Genus Actinidia. The results pointed out obvious difference among the above species, which could be considered as one of the important evidence for identification. PMID- 12572415 TI - [Improvement of product technology on prepared soypean]. AB - This paper reports the new prepared method of Prepared Soypean by purebred culture of Aspergillus niger. The technology condition is that temperture is 28 +/- 2 degrees C and comparative humidity is 95%, time of fermented culture is 15 20 days. After improvement, polluttion of other bacteria is easily avoided. PMID- 12572416 TI - [Determination on the glycosyl sequence of gypenoside A by TLC-FABMS]. AB - The glycosyl sequence of gypenoside A (A) isolated from Gynostemma pentaphyllum Makino was determinated by TLC-FABMS. The gypenoside A was hydrolysed with cellulase and the hydrolysate showed eight spots on a TLC plate, and the FABMS analysis of these spots indicated they corresponded to gypenoside A, A-Rham, A Glu, A-Rham-Glu, A-Rham-Glu, A-Glu-Glu, A-Rham-2Glu, A-Rham-2Glu in order. Two chains can be determined to be Rham-Glu, and Glu-Glu, respectively. So the glycosyl sequence of gypenoside A was identical of gypenoside XLIII. PMID- 12572417 TI - [Pharmacological actions of different species form genus Pyrola]. AB - This paper reporst the pharmacological actions and toxicities of the water extracts from Pyrola calliantha, Pyrola decorta and P. forrestiana. The results show that the actions of P. forrestiana on antiventricular fibrillation, strengthening cardiac musicular contraction and antihypoxia abitity are more stronger, but on antiinflammation and antipyretic, effects of P. decorta and P. calliantha are significantly better than that of P. forrestiana. The effects of the three species on immunization and antisepsis are similar. PMID- 12572418 TI - [Effects of n-butanol extract from Cortex Ilicis rotundae on blood pressure and heart rate in rate]. AB - It was studied that effects of n-butanol extract from Cortex Ilicis Rotundae on blood pressure and heart rate in differedt state. Results showed that the drug elicited a sgnificant depressor effect on both normal pressure and artery pressure to rise for common carotid artery blocked in rats. Of the two latter decline of percent of blood presure and active time and slowing heart rate were more significan than the former. It was same depressor effect that injected the drug into duodenum but not act to hypotensive rats. These results suggested that the drug induce significantly depressor effect and slow heart rat with the exception of hypotension. PMID- 12572419 TI - [Studies on the quality control of shangke wangshui]. AB - Shangke Wangshui is a new Chinese traditional patent medicine. Its main ingredients Coptis Chinsis Franch, Phellodendron amurense Rupr. and Gardenia jasminoides Ellis. were identified by TLC and PC. The content of active component berberine in Shangke Wangshui was determined by TLC-scanner. This mothod is simple, quick and highly reproducible and can be as a quality control standards for this preparation. PMID- 12572420 TI - [Herbal research of uigurian drug Mandragora officinarum]. AB - Mandragora officinarum L. is one of the traditional drug recorded in Uigurian document, but Uigurian doctors mistake Panax ginseng C. A. Mey. for Mandragora officinarum L. By herbal study and research of systematic botany for Uigurian drug Mandragora officinarum L., this paper considers that is doesn't substitute Panax ginseng C. A. Mey. for Mandragora officinarum L. PMID- 12572421 TI - [Studies on the characteristics of occurrence and damage for grub in field of common peony]. AB - The grub is a mixed species population that is composed of Holotrichia oblita as main part in field of Comon Peony and caused great damage to Common peony. It is critical period to control lower age larvae in July. Damage degree of grub is influenced by Common Peony varieties, soil types and growth numbers of year. PMID- 12572422 TI - [Seasonal variation on reproductive cycle of the female Chinemys reevesii]. AB - Seasonal variation in the reproductive cycle of female Chinese three-keeled pond turtles, Chinemys reevesii, was studied. Vitellogenesis began in early October and reached maximum level in early May (maximum follicle diameter was 19.2 mm). The first clutch was laid in June. Oviposition continued until August when follicular regression began. An ovarian quiescent period was evident in September. Two-three clutches are laid per year. Mean clutch size is 5. Annual reproductive potential ranges from 10 to 15 eggs. Age at maturity for females is six years. PMID- 12572423 TI - [Investigation on the resources of medicinal centipedes and identification on their commodities]. AB - Through investigation on the resources of medicinal centipedes and identification on corresponding medicinal materials, the original animals of medicinal centipedes are Scolopendra mutilans, S. multidens, S. mojiangica and S. negrocaptis. The main commercial medicinal centipede is S. mutilans. Its yield accounts for 95%. This paper arranged the retrieve tables of the morphological characters of original animals and commercial materials of medicinal centipedes. PMID- 12572424 TI - [Pharmacological study of the rhizome powder of Pinellia pedatisecta processed by different procedures]. AB - The potentiation of hypnotic activity of a barbituate and acute tocicity by the rhizome powder of Pinellia pedatisecta Schott processed by pigbile with two different traditional medicinal procedures (fermentation mixing and heating alternatively) were studied in mice. The results showed that aquatic extracts of two processed drugs could potentiate the action of hypnotics; and ethanol extracts of the drug processed with mixing and heating alterntively could significantly enhance this action, compared with that of the drug pricessed with fermentation; while no acute toxicity was measured by the two processed drugs. PMID- 12572425 TI - [Component analysis on polysaccharides in exocarp of Ginkgo biloba]. AB - This paper reports the content and component analysis on polysaccharides in exocarp of Ginkgo biloba. The results show that the content of total saccharides is 89.7%; content of polysaccharides is 84.6%; content of reductic saccharides is 5.1%; the polysaccharides are composed of glucose, fructose, galactose and rhamnose. PMID- 12572426 TI - [Supercritical-CO2 fluid extraction of the fatty oil in Terminalia chebula and GC MS analysis]. AB - The fatty oils in Terminulia chebula were extracted by supercritical-CO2 fluid extraetion, and their fatty acids and its relative contents were determined by GC MS. 12 fatty acids were identified. Palmitic acid, linoleic acid and oleic acid of them are main constituents. Compared with petroleum ether extraction method, the extraction rate from SFE-CO2 is higher and extraction time shorter. PMID- 12572427 TI - [Effects of Puerariae isoflavone on blood viscosity, thrombosis and platelet function]. AB - Puerariae isoflavone (500, 1000 mg/kg% x 7 d, p.o.) significantly lowered blood viscosity and platelet adhesion rate, inhibited thrombosis and ADP-induced platelet aggregation in rats, and showed obvious antagonism for platelet thrombosis in ADP-treated mice. PMID- 12572428 TI - [Experimental studies on immunocompetence of lophanthus roots]. AB - The effect of Lopanthus roots extract on the HI humoral immunity and cell mediated immunity in rat was investigated in this study. The result showed that the Lophanthus roots extract could enhance liver index in rat. By Lophanthus roots extract at the dosage of 2.50 g/kg in rat, the Immunoglobulin G(IgG), immunoglobulin A(IgA) and immunoglobulin M (IgM) were increased, C3 complenment exhanced and have a highly signifcance compared to the cation by the gingseng veinforce pill (P < 0.01). The Lophanthus roots extract could mildly increased effect of rosette formation and lymphocyte transformation, but no statistic sighnificance was found. PMID- 12572429 TI - [A new medicinal plant Pterocapus indicus]. AB - It has been 70 years since Ptercapus indicus Willd. was introduced into China. The red sap from its incision of the bark becomes a kind of red crystal after some hours of exposure to air, which is used as an astringent and against other diseases. Its physicochemical properties, elements and amino acids have been analysed. The structural analysis shows that the crystal is a macromolecular compound of tannic condensation and glucoside. PMID- 12572430 TI - [Pharmacognostic studies of Tibetan medicine da-li]. AB - Da-Li is a traditional medicine of Tibetan, its original plant is Rhododendron primulaeflorum Bur. et Franch and R. anthopogonoides Maxim. This paper reports the identification of Flos et Folium Rhododendri Primulaeflori on its macroscopic character, microscopical charactersitic and TLC. The comparison between the Flos et Folium Rhododendri Primulaeflori and Flos et Folium Rhododendri anthopogonoidi is also reported. PMID- 12572431 TI - [Technological study on the supercritical-CO2 fluid extraction of Curcuma longa oils]. AB - This paper first reports technological study on the extraction of Curcuma longa oils by supercritical-CO2 fluid, mainly studies the influence of pressure, temperature, flow rate of CO2 and time on the yield of the oils, determines optimum technology of extracting the oils, analyzes chemical constituents extracted, compares the oils from SFE-CO2 and SD technology, studies industrial test on the SF-CO2 extraction. The results shows pressure and temperature are more important factors of influence; the optimum technological condition is the extracted pressure 25 MPa and temperature 45 degrees C, isolated pressure I 12.5 MPa and temperature I 60 degrees C, isolated pressure II 6MPa and temperature II 38 degrees C, CO2 flow rate 9 Kg/Kg.h, and extracted time 2 h; the constituents from SFE-CO2 and SD methods are identical, but their content has some difference; SFE-CO2 technology is better than that of SD, which is high yield shorter time, etc.; the industrial production of the oils with SFE-CO2 method is available. PMID- 12572432 TI - [Isolation and identification of terpenoides and anthraquinones from Euphorbia fischeriana]. AB - In this paper, two terpenoides and one anthraquinone are isolated from the roots of Euphorbia fischeriana. Their structures are indentified by combination of physical constants and spectrum analysis as boehmerone (I), ent-16a, 17-dihydroxy atisan-3-one (II) and physcion (III). They are found in the plant for the first time. PMID- 12572433 TI - [Isolation and structure elucidation of three dihydrophenanthrenes from Ephemerantha lonchophylla]. AB - In this paper, three dihydrophenanthrenes are isolated from the Stems and pseudobulbs of Ephemerantha lonchophylla (Orchidaceae). They are identified as Iusianthridin (1), ephemeranthol-B (2) and erianthridin (3) by spectral data. PMID- 12572434 TI - [Study on polysaccharide of Lobaria kurokauae]. AB - LOK polysaccharide is isolated from Lobaria kurokauae. By chromatography, LOK is shown to be sing substance. By gas chromatography analysis, LOK composes of rhamnose, arabinose, xylose, mannose, glucose and galactose. Their molecule ratio is about 1.00:1.34:1.12:10.13:6.99:6.47. Its mean molecule weight is estimated to be 1.5 x 10(4), its main chain is joined by a (1-->4) and a (1-->6) glucosidic linkages. PMID- 12572435 TI - [Antioxidation of Pueraria lobata isoflavones (PLIs)]. AB - Isoflavones are isolated from Pueraria lobata. By studyign Pueraria lobata isoflavones (PLIs) on lipid peroxide (LPO) and superoxide dismutase (SOD) in animal tissue, this paper suggests that PLIs is an effective antioxidant. The results in vitro show that PLIs can inhibit significantly the content of LPO. The highest inbibition rate are found to be 79.7%, 84.7%, and 86.6% in liver, kidney of mice and brain of rabbit respectively. By cold induced brain injury with chalcone, the content of LPO and activity of SOD in animal blood and brain are changed. When animals are injected with PLIs, the result indicates that the PLIs can reduce significantly the content of LPO and enhance the activity of SOD in animal blood and brain, in a dose-dependent relationship. PMID- 12572436 TI - [Comparison on pharmacological functions of pheretima]. AB - Hot water extraction of Pheretima have marked actions in antipyretic, hypotensive, antiasthmatic and diuresis. Among them Guang-Pheretima and Hu Pheretima are stronger than Tu-Pheretima in antipyretic. The difference of the intensity of Pheretima in the other actions is not obvious. PMID- 12572437 TI - [A study on the quality of various species of Dalbergia]. AB - The content and main components of volatile oil and the content of totalflavone in various species of Dalbergia (DA) are studied by steam-distillation, gas chromatography-mas spectroscopy and spectrophotometry. The results show that the content of volatile oil in Dalbergia odorifera (DO) is 3.61-3.79%, and the content of total flavone is 2.51-5.82%. The content of volatile oil of other species of DA is less than 1.50%, their content of total flavone is markedly different from that of DO. PMID- 12572438 TI - [Herbological study of some medicinal plants from Viola]. AB - By herbological study, medicinal plants from Viola used as drugs are first recorded in the book "52 Diseased Preparation". The plants used as drugs in the ancient mainly are V. yedoensis, V. invospicus, V. diffusa, V. verecunda, V. davidii, V. moupinensis and V. patrinii. PMID- 12572439 TI - [A preliminary study on preparing chromosome of young cotyledon from Sophora flavescens]. AB - The present paper reports the experimental results on the preparing somatic chromosome of young cotyledon from Sophora flavescens, compared with root tip, the mitotic index of cells in cotyledon is higher, hydrolyze with enzyme and wall degradation are easely made, as the experimental material, the young cotyledon is more comeniently prepared. PMID- 12572440 TI - [The bred technique of Zaocys dhumnades]. AB - This paper reports the observational data on the laying egg, hatching, life habit, relationship between capacity for eating and growth speed of Zaocys dhumnades bred in enclosure-like garden of snake, summarizes an available breeding technique. PMID- 12572441 TI - [Identification and survey of commercial crude drug of Herba Cistanchis]. AB - This paper deals with the morphological identification, TLC analysis and survey of commercial crude drug of Herba Cistanchis (Genus Cistinche). This results provide authentic methods for the identification of Herba Cistanchis. PMID- 12572442 TI - [Pharmacognostical identification on the Chinese drug quanshen]. AB - In this paper, 5 kinds of Quanshen that original plants are Polygonum bistorta L., P. viviparum L., P. paleacum Wall., P. sphaerostachyum Meisn., P. macrophyllum D. Don. are made an initial study on the characteristics, microscopical and physichemistical sides. It provide scientific basis for its comprehensive development and ultilization, and to use in clinical. PMID- 12572443 TI - [Pharmacognostical studies on the stem and leaf of Chinese Alyxia]. AB - About the stem and leaf of Chinese alyxia, pharmacognostical studies on the character, microscopical and physichemical aspects are made, The results are certifed that the methods of determination on the character and microscopical may be adopt for stem and leaf Chinese alyxia. There are a rich amino acids and trace elements. It provide scientific basis for its comprehensive development and utilization. PMID- 12572444 TI - [The research of analgestic action of different processed products of Sparganium stoloniferum]. AB - Ananalgestic effect of different processed products of Sparganium stoloniferum in mice is observed in hot plate and acetic acid induced writhing test, The experimental results shows that water extract of S. stoloniferum and its processed products can inhibit the pain. The analgestic effect of its processed product with vinegar is the most powerful and lasting. PMID- 12572445 TI - [Physical and chemical analysis of medicinal animals of syngnathidae]. AB - The total nitrogens, amino acids, fatty acids, Inorganic elements of 12 medicinal animals of syngnathidae have been determined. It will provide scientific evidence for development of the medicinal resources and the establishment of quality standards. PMID- 12572446 TI - [Pharmacological study of cholagogic capsule]. AB - Cholagogic capsule is mainly composed of Fiuctus Cardui, Mariae, Oleum Menthae Piperilae, Radix Currmae, olia taraxaci, Oleum Hyperici. It is shown that Cholagogic Capsule increase the rat's bile secretion significantly, but bile component is not influenced effectively, the retention jaundice models of which are created by ANIT isn't influenced effectively. It can reduce the mouse pinna swelling caused by dimethythenzene and depress the metatarsal swelling induced by carragenin, the antiinflammatory effect keeps for more than 4 h. It inhibits the contraction of gallbladder smooth musele of Guinea Pig caused by Ach. The results show that cholagogic Capsule increase bile secretion, antispasmodic and antiinflammatory effect. It is suggested the therapeutic effects of Cholagogic Capsule in sand-like calcul and acute chronic cholecystitis are related to the above pharmaological effects. PMID- 12572447 TI - [Study on immunological pharmacology of keyin granule]. AB - By intragastric keyin granule at the dosage of 1.0-4.0 g/kg.d[-1] for 7-10 days in mice, this drug can enhance the phagocytic function of the macrophages of peritoneal cavity, increase the index of spleen and the clearance rate of i.v. charcoal particles, obviously inhibit the formation of mouse-specific antibody and the delayed cutaneous hypersensitivity (DCH), recover the decrease of ANAE lymphocyte in periphery blood induced by cyclophosphamide. It can also improve the index of immunological organ of immunosuppression mice. These show the immunological regulative function of this drug. PMID- 12572448 TI - [Study on the quality standard of Hippophae rhamnoides oil for pharmaceutical uses]. AB - The quality control methods of the seed oil and fruit oil of Hippophae rhamnoides are studied. The physical and chemical factors are determined and the identification and determination methods for the bioactive compnents a-tocopherol and carotenes are established. PMID- 12572449 TI - [Character of emergence for the seeds of Ephedra sinica]. AB - The experimental results on the germinating of Ephedra sinica seeds and sowing show: the seeds of E. sinica are not sensitive to illumination, but very sensitive to humidity; the effects of soil, the depth of sowing and irrigation on emergence are very notable, but not remarkable to covering which can be used to prevent measure. PMID- 12572450 TI - [Biologic characteristics and cultured methods of Palea steindachneri]. AB - Palea steindachneri is a precious animal and its carapacial bones and plastral bones are expensive Chinese medicinal materials. This paper introduces the biologic characteristics and cultured methods of Palea steindachneri. It provides a valuable information for the culture of Palea steindachneri. PMID- 12572451 TI - [Character and UV spectral identification of deer's heart]. AB - In this paper, the fresh and dried hearts of Cervus nippon and C. elaphus are identified on character, fluorescence and UV spectra. The alcohol extract of the blood of the deer's hearts has mininum absorption at UV 310 nm. PMID- 12572453 TI - [A comparative study on chemical constituents in different processed products of Dioscorea opposita]. AB - The quantitative comparisons of Water-soluble extract, alcohol-soluble extract and allantoin in crude and three processed products of Dioscorea opposita is carried out by the method recorded in Pharmacopoeia and TLCS respectively. As a result, the changes of water-soluble extract and alcohol-soluble extract during the four samples are similar. It is the hightest in the processed product stir fried with soil and lowest in the crude. But for allantoin, it is the highest in the processed product stir-fried with wheat bran and lowest in that stir-fried with no material. It is suggested that the contents between water-soluble extract, alcohol-soluble extract and allantoin are not interrelated. It shall be considered to increase the content of allantoin as a quality control index to control the quality of the processed products. PMID- 12572452 TI - [Toxic studies on various processed products of Fructus Psoraleae]. AB - Toxic studies on Fructus Psoraleae (FP) and its processed products are carried on. LD50 is 37.7 +/- 0.54 g crude drug/kg in oral administration of raw FP in comparison with 43.25 +/- 6.1/kg in that of Salt-bake one in mice. When 5.0 g/kg is given by oral for 14 days, the wet weight of testicles, preputial galnds, prostate and semianl vesicle is decreased in raw FP and its stir-bake tested groups. The wet weight of preputial glends, prostate, seminal vesicle in Lei Gong's Procession group, or prostate, and seminal vesicle in salt-steam group is also decreased to a certain extent. But no significant change on the weight of those sex organs is found in liquor-bake or salt-bake procession group, The glomerulal cell hyperplasia and cloudy swelling of renal tubules are induced by administration of raw FP in pathological examination. PMID- 12572454 TI - [Supercritical-CO2 fluid extraction of natural VE and fatty oil from the oil foot of Brassica campestris and GC-MS analysis]. AB - Natural VE and fatty oil from the oil foot of Brassica campestris are extracted by supercritical-CO2. After saponification and esterification these acidesters and natural VE in the extraction are analyzed by GC-MS. The results show that the fatty acids from SFE-CO2 contain 10 components which of oleic acid, linodeic acid and linolenic acid are main components, linodenic acid and linoleic acid of them are increased signifantly, at the same time, natural VE can be extracted from the materials and the rate of extraction gets to 95%. PMID- 12572455 TI - [Studies on the chemical constituents of Litsea glutinosea]. AB - Two crystalline substances are isolated from aerial parts of Litsea glutinosea (Lour.) C. B. Rob. and are identified as a-phenylcinnamic acid and p-cumarric acid. PMID- 12572456 TI - [The constituents of the volatile oil from Achillea wilsoniana]. AB - The chemical components of essential oil from herba of Achillea wilsoniana is identified by GC-MS techniques, and 35 compounds are identified, mainly containing beta-pinene, limonene, t-beta-farnesene, delta-cadinene, alpha cadinol, 1,4-dimethy-7-ethylazulene. PMID- 12572457 TI - [The effect of cimeiguo on micronuclei in mice induced by cooking lampblack]. AB - The effect of Cimeiguo on micronuclei in mice induced by cooking lampblack is studied. The results show that the number of micronulei induced by cooking lampblack are increased sighificantly (P > 0.01) with dose-response relationship and inhibited by Cimeiguo. PMID- 12572458 TI - [Antianoxic effects of Dracocephalum tanguticum on brain of mice]. AB - The aqueous extracts i.p. of Dracocephalum tanguticum Maxim (DtM), a Tibetan medicine, prolong sighificantly decapitaion-induced gasping duration in mice and the survival time of mice ligated bilateral carotid (P > 0.01). DtM prolong also the primal gasping in mice after injecting the air into the coccygeal vein (P > 0.01) and the survival time in mice poisoned by Lidocaine (P > 0.05). In addition, DtM is possessed of protecting against the pathologic lession of brain in rats under stimulating high altitude hypoxic condition. The results suggest that DtM has a antianoxic effects on mice. PMID- 12572459 TI - [Cultivation of Polygonum multiforum]. AB - The experiments of some cultivated medthods of Polygonum multiforum show that the direct sowing of seeds is better than the cuttage, the index of reproduction and yield are higher, and the cultivated method can decrease expense. To cultivation period of 5 years, high yield and excellent qualty is available. PMID- 12572460 TI - [Reproduction and cultivation of Neocheiropteris palmatopedata]. AB - Neocheiropteris palmatopedata is a rare medicinal plant which only distributes in the boundary among Guizhou, Yannan and Sichuan. The paper reports cultivation methods with using rhizome and spores of N. palmatopetata as reproduction materials. PMID- 12572461 TI - [Survey on medicinal resources of Syngnathidae]. AB - A survey on medicinal resources shows 12 species belonging to 7 genera of Syngnathidae are used as traditional Chinese medicine and some problems about cultivated Haima have been discussed. PMID- 12572462 TI - [Identification of deer's foetus and its mixed drugs and flase drugs]. AB - In this paper, the traditional Chinese Materia Medicas--the foetuses of sika deer, red deer and the one of sheep and OX are compared and identified on characters and dissecting characteristices. The comparion and identification table about charaeters and commodity shape drawings is attached. PMID- 12572463 TI - [Study of supercritical-CO2 fluid extraction in extracting essential oils of Amomun tsao-ko]. AB - Essential oils from Amomum tsao-ko are extracted by supercritical-CO2 fluid and the oils are analyzed by GC-MS. 1, 8-cineole, trans-geraniol eicosatrienoic acid methylester and d-nerolidol of them are the major constitutents, 12 constitutents of them are first obtained from the plant. PMID- 12572464 TI - [Determination of oridonin in Rabdosia rubescens by TLC-scanning method]. AB - We have determingd the content of oridonin in Rabdosia rubescens Hemsl by TLC scenning method. This method is proved to be simple, fast, precise and sensitive, and the result is stable, repetition property is good. Its rate of recovery is as high as 98.03 percent. PMID- 12572465 TI - [Spectrophotometry analysis of berberine chloride in xianglian jiaowan]. AB - After isolation of column chromatograph with organic solvent containing acid water, the content of berberine chloride in Xianglian Jiaowan (a Chinese drug) is determined by spectrophotometry. The analysis method is specific, sensitive and accurate, recovery is 99.8% and RSD is o.78%. PMID- 12572466 TI - [Inhibition of 5 compounds from Bidens bipinnata on leukemia cells in vitro]. AB - In vitro, inhibition of 5 compounds from Bidens bipinnata on two leukemia cells HL-60 and V397 is presented by determination of MTT living cell in this paper. The result show that the compounds have the action of inhibition, IC50 on V937 < or = 60 micrograms/ml. PMID- 12572467 TI - [Studies on hypoglycemic action of Rheum emodi]. AB - The hypoglycemic action has been studied with the 95% ethyl alcohol extract and stilbene glycoside E (exists in a large amount), which are obtained from Rheum emodi. The experimental result shows us the utilization value of R. emodi. In the meanwhile, as tell us how to use the sect Rheum resources. PMID- 12572468 TI - [Herbological study of 4 Chinese herbs in Ardisia and "yiedihong"]. AB - Herbological study are given to the original plants of "Zijinniu", "Pingdimu", "Xiaoqing", "Duanjiaosanlang" and "Yiedihong" recorded in the herbal books. The research shows that they are not the same plant with different names. Also, "Yiedihong" doesn't belong to Ardisia. PMID- 12572469 TI - [Enzynology of infected part of red skin ginseng]. AB - By means of polyacrylaminde gel eletrothoresis and thin layer scanning, the paper semi-quantitatively studies some relation between component and activity of isoenayme of Ginseng infecting red skin desease. The results show activity of esterase increases 100% than that of normal Ginseng; activity of polyphenol oxidase increases 60-30% and represent new isoenzyme bands in the early infection in juxtapose transolant experiment. PMID- 12572470 TI - [Relationship between variations of temperature and humidity with reproduction of medical meloids]. AB - This paper reports the relationship between variations of temperature and humidity with reproduction of two medical meloids (Mylabris phalerata, M. cichorii). The results show: for the copulation of two meloids, the suitable temperature is 22-28 degrees C, humidity 26-90%; for the suitable temperature and humidity of their oviposition, M. phalerata 26-28 degrees C and 60-68%, and M. cichorii 23-26 degrees C and 72-78%, respectively; for incubation, M. phalerata is 26-28 degrees C and 70-75% respectively, and M. cichorii must undergo the high and lower variation of temperature, the efficacious accumulative temperature must get of 1680.85 day. degrees C. PMID- 12572471 TI - [Investigation and identification of original plants of Herba Dianthi produced in Shandong]. AB - The original plant of Herba Dianthi in Shandong are divided into 4 species and 2 varieties by investigation and classified identification. Of them Dianthus shandongensis J. X. Li et F. Q. Zhou is a new species and D. chinensis L. var. versicolor (Fich. ex Link) Y. C. Ma is a new recorder in Shandong. D. chinensis L. is the most important source of Herba Dianthi in Shandong and the D. chinensis L. var. versicolor and D. chinensis L. var. liaotungensis Y. C. Chu, occasionally is D. shandongensis, D. superbus L. and D. longicalyx Miq. Haven't grown into commercial medicinal materials. PMID- 12572472 TI - [Pharmacognostical identification of Harba Swertice mileensis and its substitutes]. AB - The present paper provides scientific basis for "Qingyedan" (Harba Swertiae Mileensis) through comparison studies of characteristics, histological morphology, and physico-chemical analysis with its three substitutes which are Swertia punicea Hemsl, S. nervasa Wall, and Halenia elliptica D. Don. PMID- 12572473 TI - [Identification of herba hedyotis diffusae and its confused material Herba Hedyotis pinifoliae]. AB - This article has reported the results of the study on the outer properties, inter organization and TLC of Herba Hedyotis Diffuse and its confused material Herba Hedyotis Pinifoliae. The results show the two have obvious distinction and can not be confusediy used. PMID- 12572474 TI - [Polyacetylene compounds from Panax notoginsenoside]. AB - In this paper, two knowm polyacetylene compounds, falcarindiol(1) and panaxytriol(2), are isolated from panax notoginsenosides. PMID- 12572475 TI - [Separation and identification of chemical compositions from cultivated Belamcanda chinensis]. AB - In this paper, two compounds, isoflavone III and IV are isolated. On the bases of physical, chemical and spectral evidences, the structure of composition III is tectoridin, and composition IV is tectoriginin. PMID- 12572476 TI - [The regulation effects of Angelica sinensis on the erythrocytic immune function and IL-2 in mice]. AB - The effects of Angelica sinensis solube extract on erythrocytic C3b receptor rosette rate and immune complex rosette rate and activities of interleukine-2 (IL 2) in mice are observed in this article. Angelica sinensis can significantly enhances erythrocytic C3b recepor rosette rate and IL-2 activities. The regulating actions of Angelica sinensis on erythrocytic C3b recepor rosette rate and IL-2 activities may be the direct effects. PMID- 12572477 TI - [Effects of extract of Cortex Ilicis Rotundae on cardiovascular pharmacology]. AB - In thes paper, the effects of Cortex Ilicis Rotundae (CIR) on Cardiovascular Pharmacology are studied. The results show that ethyl alcohol extract of CIR can reduce coronary blood flow, weaken myocardial contractility, slow down heart rates, prolong survival time of mice and prevent arrhythmias, the roles of CIR decoction are similar to ethyl alcohol extract, but it is not markable on heart rates and myocardial contractility. PMID- 12572478 TI - [Clinial study of "qingjunyin" detoxification for the treatment of heroin addicts]. AB - To evaluate the efficacy of treatment of heroin addicts (n = 100) by "Qingjunyin" (QJY) detoxification (10 days program). Methadone detoxification (10 days program) group (n = 50) and colonidine treated group (n = 50) serve as controls. QJY group is given 180 ml P. O daily. The dosage of the two comparatine groups is given according to Documentation, the method of abserving the treatment protocols and detoxification standard is according to current regulation. The scores of abstinence syndrome in QJY detoxification group are lower than those in colonidine treated group in the first three days of potocol, but this difference disappear in the late stage of treatment. While QJY detoxification is effective as methadone detoxification in the control of abstinece syndrome during the first five days of treatment but the difference in the scores of abstinence syndrome between QJY and methadone group is observed during the late five days of protocol. QJY does not result in potential dependence and has definite curative effect in the treatment of heroin addiction. PMID- 12572479 TI - [Study on the stunted culture of trees of Scaphium lychnophorum]. AB - Taking these cultural practices, grafting by the ingraftment of side branch on the place of flowering with the dwarf stock, tip pruning on time to make branches growing and rational pruning etc can make the trees of Scaphium lychnbophorum dwafing effectively. The stunted trees are only high 3-5 m and their branches are only high about 1 m from the ground. Some of them first flower until five years after they are cultivated. PMID- 12572480 TI - [The preventing and control of ginseng Rhizoctonia solani with nymexazol]. AB - This paper reports the effect on nymexazol protecting the Rhizoctonia solani of Panax ginseng. The protective effect is in 80-95%. The drug can substitute quintozene and fenaminosulf in production. PMID- 12572481 TI - [Observation on the life habit of Epicata aptera]. AB - Epicata aptera reproduces one generation every year. Adults arise in the last ten day period of May, mate and lay egg in the first ten-day period of June. They mate and lay egg 1-3 sequence every year, lay egg 108-269 grains every sequence. Period of their laying egg undergoes 48-78 days. Larva have six age period. They take false pupas live through the winter. Complete generations undergo 348-394 days. PMID- 12572482 TI - [The morphological and commercial identification on medicinal materials "hailong"]. AB - The morphological characters and commercial idendification on medicinal materials named "Hailong" has been reported. The results of identification of 48 commercial samples indicated "Hailong" is adulterated with Halicampus koilomatodon, Trachyrhamphus serratus and Microphis boaja, Solenognathus guntheri besides three species accepted by Pharmacopoeia. PMID- 12572483 TI - [Pharmacognostic identification of the root bark of Broussonetia papyrifera, an adulterant of cortex mori]. AB - The article reports the morphological characters, the histological structure, TLC and UV spectrum identification for the adulterant of Cortex Mori, the root bark of Broussonetia papyrifera. The results show that they can be distinguished each other. PMID- 12572484 TI - [Isolation and properties of a novel fibrinolytic enzyme from an earth worm]. AB - A novel fibrinolytic enzyme is isolated from one species of Pheretima by means of homogenizing, extracting with an extractive agent, precipitating with ammunonium sulfate, ultrafiltration and chromatography. The enzyme consists of a single chain with an M. W. of 22,000. It can not only dissolve human thrombi and fibrin directly and strongly, but also activate human plasminogen. The enzyme shows little toxic and side effects in animal tests. The activity, purity and etraction rate of the enzyme in this report are all very high. PMID- 12572485 TI - [Studies on the effective constituents of arresting bleeding of Schizonepeta tenuifolia]. AB - Schizonepeta tenuifolia (Benth.) Briq fried with carbon is used in various bleeding diseases in order to further investigate the effective constituents of arresting bleeding in Schizonepeta tenuifolia(benth.) Briq and to finish the quality standard of its capsule. We isolate the chemical constituents from Schizonepeta tenuifola(Benth.) Briq fried with carbon and study its action of arresting bledding and assay the effective constituents of arresting bleeding in capsule. PMID- 12572486 TI - [Effects of volatile oil from Rheum palmatum on immunologic function in mice]. AB - The effects of volatile oil from Rheum palmatum (RVO) on immunologic function in mice are studied. The results show that the delayed type hypersensitivity (DTH) induced by 2, 4-dinitrochlorobenzene, lymphocytes transformation of (3H)-TdR induced by PHA, clearance rate of ivcarbon particles, phagocytic function of peritioneal macrophage and production of hemolysin induced by SRBC are markeyly increased with RVO given by oral administration or inhalation. PMID- 12572487 TI - [The antibechic, expectorant and antiasthmatic actions of Cynanchum komarovii]. AB - The water and ethanol extracts of Lao Gua Tou (Cynanchum komarovii) is effective on relieving cough caused by the irritation of ammonnia water in mice. The extract can increase the secretion of repiratory tract and promote expectoration, also alleviate asthma induced by histamine and acetylcholing chloride in guinea pigs. PMID- 12572488 TI - [Herbalogical studies of the medicinal plants on Rubus]. AB - This paper deals with the investigation of the medicinal plants of Rubus recorded in the herbalogical works of the past dynasties. The substitutes and false drug of Fu Penzi are also investigated. The result show that Fu Penzi recorded in Materia Medica shall be R. coreanus, and Pong Le be R. lambertianus in modern times. PMID- 12572489 TI - [Comparative test on spawn culture materials of Poria cocos]. AB - It saves much time, work and timber when Poria cocos spawn is made from cottonseed hulls and pine sawdust. Compared with the yields from pine wood chip, the difference is not significant. PMID- 12572490 TI - [Pharmacognostical identification of niuxi from Guizhou Province]. AB - In this paper, 7 kinds of Niuxi from Guizhou Province have been studied on their pharmacognostical characters, histological characters and effective constituents and the difference of species have been found based on the experimental results. PMID- 12572491 TI - [Micromorphological identification on 3 species of medicinal plants for genus Peganum]. AB - This paper reports the micromorphological features of the leave, pollen grains, petals and seeds of 3 species of medicinal plants of Genus Peganum. The results can supply basis for the identification of the medicinal material. PMID- 12572492 TI - [Determination of ampelopsin and myricetin in Ampelopsis cantoniensis]. AB - Ampelopsis cantoniensis (Hook. et Arn.) Planch. is a Chinese herbal medicine rich in flavonoids. A method to determine flavonoids in the herbal was studied. After the herbal was extracted in hot water, Ampelopsin and myricetin in the extract were separated by polyamide column with eluent of ethanol-water and measured by spectrophotometry. As results, 100 g of the dry herbal contains 25.2 g of ampelopsin (RSD = 1.7%, recovery = 97.0%) and 1.77 g of myricetin (RSD = 3.3%, recovery = 103%). PMID- 12572493 TI - [Content determination of total coumarin of qian-hu]. AB - The total content of coumarin in Qian-Hu was determined by UV spectrometry. The 25 samples which derived from different places of production were included eleven varieties. The results were found to have max = 322 nm, 99.18% of recovery rate and 2.08% of RSD. PMID- 12572494 TI - [Studies on chemical constituents of the essential oil from Angelica formosana]. AB - Essential oil from Angelica formosana Boiss was obtained by steam distillation. The chemical components of the oil were examined by means of capillary gas chromatography and gas chromatography-mass spectrometry. 23 of the 62 separated constituents were identified. The content of each component identified was determined by area normalization method. Of these, the content of 11 compounds was higher than 1%. PMID- 12572495 TI - [Supercritical-CO2 fluid extraction in extracting volatile constituents from Juniperus formosana]. AB - Volatile oils from the leaves of Juniperus formosana Hayata were extracted by supercritical-CO2 fluid. These constituents were analysed by GC-MS. The result showed that the oils from CO2-SFE contain 29 components, 21 of the them were identified, which are first obtained from the plant. PMID- 12572496 TI - [Hemolysis of Scolopendra toxins]. AB - The hemolysis of toxins from alive Scolopendra subspinipes mutilans, medicinal material of Scolopendra subspimipes mutilans and S. multidens have been compared. The result shows that all the toxins have hemolytic activity. The hemolytic activity of the toxin from the medicinal materials of S. subspinipes mutilans is obviously lower than that from alive ones, and that from fresh medicinal materials are twice as high that from old ones, and that from S. multidens is higher than that from S. subspinipes multilans. PMID- 12572498 TI - [Herbological investigation on the traditional Chinese medicine zhimu]. AB - In this paper, herbological investigation was carried out into a large of historical harbal books about Zhimu, Rizoma Anemarrhenae, especially about its orgin, method of preparation. The description in "A Record of Chinese Medica" was partly revise. PMID- 12572497 TI - [Preliminary study on the pharmacological action spicatus]. AB - In this report the pharmacological action of Spicatus was studied. The results insicated that it had diureric, antibiotic and anti-inflammatory effects, yet had Iittle toxic side-effect. It had significant inhibitory effect on crofon oil caused mice ear swell. It also had marked diuretic effect in orcinary rats, but had Iittie effect on uric pH the rats. It exhibited certain inhibition of Staphycoloccus aureus, Eschrichia coli and Pseudomonas aeruginosa in vitro. The maximum tolerable dose test in mice showed no marked toxic effect, LD50 > 80 g/kg. PMID- 12572499 TI - [Quick propagation and intermediate test on Anoectochitus formosanus]. AB - This paper reports the technique of quick propagation of Anoectochitus formosanus on tissue culture and intermediate test on factory production. The results may provide a reference for its further study and development. PMID- 12572500 TI - [Studies on the pharmacognosy of Cortex Illicii and its adulterants]. AB - The traditional Chinese Medicine Cortex Illicii and its adulterants were studied on textual identification, botanical origin, morphological and histological characters. The TLC and UV spertra methods were established to separate and estimate Difengpin, Magnolol and beta-sitosterol in Cortex Illicii and its products. PMID- 12572501 TI - [The SEM observation of the medicinal materials "hailong"]. AB - The SEM observation of the medicinal meterials "Hailong" have been carried out. The results showed there are significantly difference among 7 species "Hailong". PMID- 12572502 TI - [Morphological histological and UV identification of radix Linderae]. AB - The present paper reports the differences of the root tuber, cylindrical root and stem of Lindera aggregata (Sims.) Kosterm. on the external characteristics of crude drug, internal structures and UV-identification. PMID- 12572503 TI - [Comparative study of the constituents from 10 Rhodiola plants]. AB - The comparison of the components in 10 species of Rhodiola by GC method showed that sucrose and peak A, a unknown constituent, exist in all of 19 samples collected in the east area of Qinghai Province and in high quantities. However, salidroside, a known active component from the genus, is found from only 5 species with the quantity of more than 0.3%. There are 5 plants containing lotaustralin, a known oral toxic compound, and with the highest quantity in the plant of Rh. kirilowii. The content of the two compounds above varies as plants and habitats are different. PMID- 12572504 TI - [Study on leaching ratio of lead in the process of decocting medicinal herbs]. AB - The content of lead in the raw material and the water leaching solution of 56 kinds of traditional Chinese medicine were determined by AAS method, the leaching ratio of lead in the process of decocting medicinal herbs was obtained. PMID- 12572505 TI - [Studies on the antilipid peroxidation of nine sorts of Chinese herbal medicines with the function of protecting liver]. AB - The antilipid peroxidation of nine sorts of Chinese herbal medicines with the function of protecting liver, including Salivia miltiorrhiza, Hypericum japonicum, Scutellaria baicalensis, Callicarpa cathayana, Chrysanthemum indicum, Paeonia latiflora, Lysimachia christinae, Ligustrum lucidum (L1), Patrinia villosa (Pv) on hepatic homogenate of rat were tested. It was found that all tested medicines showed inhibition with dose-effect relationship, the inhibitory rate of L1 and Pv were lower than the other's. All results showed that these Chinese herbal medicines have strong antilipid peroxidation and are natural oxidation inhibitor. It was suggested that their function of protecting liver and others have relationship with the antioxidation. PMID- 12572506 TI - [Protective effect of paeonol on repeated cerebral ischemia in rats]. AB - Protcective effect of paeonol was observed in the repeated cerebral ischemia reperfusion model in rats. Paeonol (100, 50 mg/kg x 7 d i.p.) could inhibit the increase of Ca2+, the decrease of superoxide dimutase (SOD) activity and the content of malondialdehyde (MDA), improve Ca(2+)-ATPase activity in ischemia brain tissue. The results showed that paeonol could protect cerebral ischemia by inhibiting accumulation of Ca2+ and production of oxygen free radical. PMID- 12572507 TI - [Studies on the analgesic and antiinflammatory action of radix Linderae extract]. AB - Both of aqueous extract and alconol extract of Radix Linderae at a dose 5 g/kg and 10 g/kg could length obviously pain threshold in mice in hot plate test. The samples at a dose 20 g/kg could inhibit significantly the writhing frequency induced by potassium antimony tartate in mice, also antagonize the swelling of ear induced by inflammatory agent and decrease the swelling rate. The component further isolated from the plant could antagonize the swelling of rat toes induced by carrageenin. PMID- 12572508 TI - [Studies on the distribution and the control decision of white grub population in field of Paeonia lactiflora]. AB - The present paper showed the result of accumulated distribution pattern of the mixed population of white grubs [Holotrichia oblita (Faldermann)] in the field. The x2-test indicated that the numbers of grub were 2.5-8 per squar-meter, which agreed to negative binomial distribution. The accepted and refused lines of sequential sampling were calculated in different occurrence degree. PMID- 12572509 TI - [Preliminary studies on grafting of Santalum album]. AB - With the purpose of propagating high production Clone of Santalum album, the best season and practical method of grafting, and the selection of shoots for scion are studied. The preliminary results show: The best season for grafting in Guangzhou District occurs from June to October, when the daily mean temperature is over 25 degrees C, the side graft is recommedable; the scion from 1-5-year old young trees is much in favor for grafting than that from adult trees. In the right condition, side grafting of Santalum album has had up to 80 percent success rate. PMID- 12572510 TI - [The morphological and commercial identification on Haima]. AB - The morphological characters and commercial identification on Haima has been reported. The results of identification of 60 commercial samples indicated that all of Haima was the species accepted by Pharmacopoeia. Hippocampus kelloggi, H. histrix, H. kuda and H. trimaculatus were mainly used in Hainan, Guangdong and Fujian Province; H. japonicus was mainly used in Shandong and Liaoning Province; Imported Haima was H. histrix. PMID- 12572511 TI - [Pharmacognostical identification of Anoectochilus roxburghii]. AB - This paper reported the pharmacognostical identification on characters, microscopic characteristics and TLC of the Anoectochilus roxburghii (Wall.) Lindl., and compared with different resouce samples. The results may provide reference for its identification. PMID- 12572512 TI - [Constituent analysis of the essential oil in leaves of Torreya nucifera]. AB - The constituents of the essential oil in leaves of Torreya nucifera (L.) Sieb. et Zucc. (EOLTN) in China were firstly analysed by GC-MS-DS. 68 of the 71 separated constituents in EOLTN were identified. The total contents of identified constituents accounted for 96.364% of EOLTN. PMID- 12572513 TI - [The antitumour activity of Sinopodophyllum emodi]. AB - This paper has shown the cytocidal effects of alcohol extracts of roots and rhizomes of Sinopodophyllum emodi (Wall.) Ying on human erythroleukemia K562 cells, leukemia L1210 cells, leukemia L7712 cells in vitro, using microculture method for 24 hours. The survival rates on K562 cells in the final concentrations of 10.0, 5.0, 2.5, 1.0 micrograms/ml of the extracts of Sinopodophyllum emodi (WA11.) Ying were 44.17%, 47.63%, 64.43%, 79.57% respectively. The survival rates on L1210 and L7712 cells were 42.84%, 50.73%, 63.21%, 75.10%, and 39.76%, 46.36%, 61.42%, 75.24% respectively. LD50 of the extracts singly i.p. in mice was 76.3 60.6 mg/kg. In vivo, the growth of transplanted mouse tumours (EAC, U14 and Hc) was inhibited by the extracts of Sinopodophyllum emodi (Wall.) Ying with inhibitory rates 42.2%, 38.8%, 41.5% (14.0 mg/kg); 37.8%, 33.3%, 35.6% (7.0 mg/kg). PMID- 12572514 TI - [Pharmacological studies of luohanguo yanhou tablet]. AB - Luohanguo Yanhou Tablet possesses the action of cotton ball granuloma of mice and the auricular edema induced by xylol, and also inhibiting the swelling of rat hind paw induced by carrageenin. The tests of mice writhing induced by acetic acid showed that the preparation had obvious analgesic action. It also possesses the obvious bacteriostatic action in vitro on five kinds of strains. PMID- 12572515 TI - [Preparation of no-sugar huazhen granules]. AB - This article introduces the preperation, especially its technologies of spraying, drying and making grain of no-sugar Huazhen Granules (composed of Flos lonicerae, Radix Bupleuri, etc.). Ideal grain can be obtained, the capacity of taking can be reduced greatly. PMID- 12572516 TI - [Cultivated technique of high yield for Polygonatum officinale]. AB - By studying cultivated materials, hided crops, cultivated way, applied fertilizer about the cultivation of Polygonatum officinale, put forward following the methods of cultivation of P. officinale. Choose rhizome segments with top buds as plant materials, adopt level rectangular pieces of land in a field width 100 cm, cultivate late-maturing corn as hiding shady, adopt density of 1.5-2.0 ten thousand plants/mu, apply N and P fertilizers in the growth. PMID- 12572517 TI - [Influence of leaf-picking on effective constituents of the Astragalus mongholicus roots]. AB - After the application of the orthogonal design, experiments show that the proportion and time of leaf-picking have a great influence not only on the total content of 17 kinds of amino acids but also on that of Astragaloside. If the proportion is large and the time is delated, both contents of 17 kinds of amino acids and Astragaloside will be reduced accordingly compared to the control group. There is less influence on the contents if the time of leaf-picking is set on June 15 each year and the picking proportion is 1 out 3. PMID- 12572518 TI - [Observation on pollen morphology of six species of Typha (Puhuang) by SEM]. AB - The pollens of 6 species of Typha (Puhuang) have been observed by SEM. The morphological characters of the pollens are described. PMID- 12572519 TI - [The research on analgestic and anti-inflammatory action of different processed products of Achyranthes bidentata]. AB - Analgestic effect of different processed products of Achyranthes bidentata in mice was observed in hot plate and acetic acid induced writhing test. The experiment results showed that water extract of Achyranthes bidentata and its processed products could inhibit the pain. The analgestic effect of its product processed with wine is the most powerful and lasting. The results showed that Achyranthes bidentata could obviously inhibit the ear inflammation of mice which was led by croton fruit oil. The anti-inflammatory effect of its product processed with wine is the most powerful. PMID- 12572520 TI - [Studies on the chemical constituent of Ephedra likiangensis]. AB - Six crystalline substances were isolated from the stem of Ephedra likiangensis Florin and were identified as ephedrine (I), beta-sitosterol (II), palmitic acid (IV), stearic acid (V), cerotic acid (III) and beta-daucosterol (VI). PMID- 12572521 TI - [Analysis of the chemical constituents of volatile oils of Metasequoia glyptostroboides leave]. AB - The chemical constituents of volatile oils of Metasequoia glyptostroboides leave were analyzed by GC-MS-DS. 27 constituents were identified, alpha-pinene (70.65%) and caryophyllene (10.38%) of them are main components. PMID- 12572522 TI - [Anti-inflammatory effect of total flavonoidal glycosides of Swainsona salsula]. AB - Total flavonoidal glycosides (SSTFG) was extracted from Swainsona salsula (Pall.) Taub.. The anti-inflammtory effect of SSTFG was investigated. The results indicated that SSTFG inhibited markedly dimethylbenzene-induced inflammation in the ears of mice and albumen--caused swell in the feet of mice. SSTFG also exhibited a significant antagonism against increase in permeability of the peritoneal capillary of mice caused by acetic acid. PMID- 12572523 TI - [Comparision on the effects of ALI aeute liver injury of three species of concha haliotidis in mice]. AB - The liver injury mice model with Haliotis ruber (Leach), Haliotis disus hannai Ino and Haliotis lacvigata (Donovan) was studied. The results indicated three species Haliotidis could versus carbon tetrachloride-induced ALI aeute liver injury in mice. Comparision of reduced serum glutamic-pyruvic transaminass (SGPT) activity showed Halitis disacus hannai Ino > Haliotis Lacvigata (Donovan) > Haliotis ruber (Leach). The Liver starch was determined. It is suggested protection liver effect of Haliotis discus hannai Ino and Haliotis lacvigata (Donovan) are most powerful. PMID- 12572524 TI - [An observation of the percutaneous absorption of total alkaloids and terpenoid lactones of Tripterygium wilfordii]. AB - The main ingredients of Tripterygium wilfordii are total alkaloids and terpenoid lactones. The authors studied the percutaneous absorption of these compounds on mice by means of ultravillet spectrophotometry. The results showed that these alksloids and lactones could pass through the skin of the mice. The rate of 12 hour percutaneous absorption was 13.40% for the former and 17.60% for the latter. After the application of Tripterygium wilfordii adhesive plaster to the human skin for 12 hours, the two ingredients were absorbed 17.34% and 22.13% respectively. This suggests that Tripterygium wilfordii can really be administered through the skin. PMID- 12572525 TI - Synthesis and evaluation of anticonvulsant activities of some bis Mannich bases and corresponding piperidinols. AB - Some acetophenone derived bis Mannich bases (B1-B5) and piperidinols (C1, C4), which are the structural isomers of B1 and B4, and also quaternary piperidine derivative C6 were synthesized and studied for anticonvulsant activity. Of the compounds, C6 was reported for the first time. Chemical structures of the compounds were confirmed by UV, IR, 1H-NMR, 13C-NMR, mass spectra and elemental analysis. Their anticonvulsant activities were determined by maximal electroshock (MES), subcutaneous metrazol (scMet) tests and rotarod test for neurological deficits. According to the activity studies, B2, B4, C1 and C4 derivatives were found to be protective against MES at 30 mg/kg and above. B1, B2, B3, B4, C4 and C6 derivatives were found to be protective against scMet. at different dose levels ranging from 30 to 300 mg/kg. Since no neurotoxicity was detected for the compounds B4 and C4, they seem to be candidate compounds for further synthesis and in vivo studies for their potential anticonvulsant activity. PMID- 12572526 TI - Sufentanil-related respiratory depression and antinociception in the dog. Mediation by different receptor types. AB - The mu-receptor purportedly is considered the site responsible for the mediation of opioid-related respiratory depression. However, there is no equivocal understanding whether the same site is also responsible for antinociception. For blockade of effects, the selective mu-antagonist beta-funaltrexamine (CAS 72782 05-9, beta-FNA) was given intracerebroventricularly (i.c.v.) prior to increasing doses of sufentanil (CAS 60561-17-3) (3, 6 and 12 micrograms/kg) in the conscious dog. This was followed by the selective delta-antagonist naltrindole (CAS 111555 53-4) (160 micrograms/kg). After one week, using the same dosages and the same animals, saline instead of beta-FNA was given i.c.v., again followed by sufentanil and naltrindole. Arterial blood gases (paO2, paCO2) were used to demonstrate respiratory impairment while somatosenory-evoked potentials reflected sensory blockade. Maximal depression of paO2 was 73.9 with and 55.0 mmHg without beta-FNA, while paCO2 rose to 44.7 without and to 35.0 mmHg with beta-FNA (p < 0.005). In the evoked potential, maximal depression was 39.1% with and 92.7% without beta-FNA (p < 0.005). Naltrindole reversed residual hypoxia, however, not hypercarbia or amplitude reduction of the evoked potential. For regulation of paO2, a mu-delta-receptor interaction is postulated while paCO2 and sensory blockade are affected solely by the opioid mu-site. PMID- 12572527 TI - Topical treatment of rhinosinusitis with fusafungine nasal spray. A double-blind, placebo-controlled, parallel-group study in 20 patients. AB - In a monocenter, placebo-controlled, double-blind, parallel-group study, 20 patients with acute symptoms of rhinosinusitis were treated with either fusafungine (CAS 1393-87-9, Locabiosol Dosier-Spray) (n = 10) or placebo nasal spray (n = 10). One patient from the placebo group was withdrawn from the study on the day of inclusion for noncompliance reasons. At the beginning of the 2-week treatment period, absence of an acute exacerbation of sinusitis with, e.g., opacity or fluid in any of the sinuses, was documented by computed tomography. Efficacy of treatment was assessed using objective measurements (e.g. sonography, rhinomanometry, acoustic rhinometry, and endoscopy) as well as scores recorded by the investigator on 6 consecutive visits before and after 1, 3, 7, 10, and 14 days of treatment, and by the patients on a diary card. Drug safety was evaluated on the basis of twice-daily assessment of general well-being and side effects. Statistical analysis of the data evidenced a positive effect of fusafungine as early as by the first 24 h of treatment (reduction of symptom score D0-->D1: p < 0.01), which was not seen in the placebo group (p = 0.2174). In the final assessment, both investigator and patients rated global efficacy better with fusafungine. In some instances, the intergroup difference achieved statistical significance (patient diaries, p = 0.0342). Side effects such as reactions at the application site mainly occurred in patients in the placebo group, who rated tolerability markedly worse than patients taking fusafungine (p = 0.0304; patient diaries, p = 0.0170). PMID- 12572528 TI - 1-ethyl and 1-propylazacycloalkan-2-one ester prodrugs of ketoprofen. Synthesis, chemical stability, enzymatic hydrolysis, anti-inflammatory activity, and gastrointestinal toxicity. AB - Six ketoprofen (CAS 22071-15-4) alkylazacycloalkan-2-one ester derivatives (I-VI) were synthesized and evaluated for their anti-inflammatory, analgesic, and ulcerogenic activities after oral administration. Furthermore these derivatives were assayed to determine in vitro their stability in pH 7.4 phosphate buffer and in simulated gastric fluid (pH 2.0 buffer) and their susceptibility to enzymatic cleavage in rat plasma. All the prodrugs showed a good stability both in pH 7.4 phosphate buffer and in pH 2.0 buffer, and they were readily hydrolyzed by rat plasma. Esters I-VI showed an anti-inflammatory activity, determined as the percent of inhibition of carrageenan-induced edema, similar to that of ketoprofen, although at higher doses. They were significantly less irritating to the gastric mucosa than the parent drug. In the mouse acetic acid induced writhing assay, the prodrugs exhibited, following acute administration, a good analgesic activity. PMID- 12572529 TI - Effect of tolterodine on the anticoagulant actions and pharmacokinetics of single dose warfarin in healthy volunteers. AB - This randomized, double-blind, crossover study investigated the potential effects of tolterodine ((R)-N,N-diisopropyl-3-(2-hydroxy-5-methyl-phenyl)-3 phenylpropanamine, CAS 124937-51-5), an antimuscarinic agent for the treatment of the overactive bladder, on the anticoagulant actions and pharmacokinetics of single-dose warfarin (CAS 81-81-2) in 20 healthy male volunteers. In terms of study design, volunteers randomly received oral tolterodine L-tartrate (2 mg twice daily) or matching placebo for 7 days, with a single oral dose of warfarin (25 mg) administered on day 4 of each treatment period. R-(+)- and S-(-)-warfarin pharmacokinetics were estimated from plasma levels measured up to 96 h post-dose, in conjunction with assessment of prothrombin time and factor VII activity. Pharmacokinetics of tolterodine and its active 5-hydroxymethyl metabolite ((R) N,N-diisopropyl-3-(2-hydroxy-5-hydroxymethyl-phenyl)-3-phenylpropanamine; 5-HM), in the presence and absence of warfarin, were also determined. Relative to placebo, tolterodine had no discernible effect on the anticoagulant actions of warfarin. Point estimates of the tolterodine: placebo ratios for prothrombin time and factor VII activity were 1.00 (90% confidence interval [CI]: 0.91-1.10) and 0.91 (90% CI: 0.83-0.99), respectively, consistent with equivalence. No clinically significant changes in the pharmacokinetics of R-(+)- and S-(-) warfarin were noted. Serum concentration-time profiles and the pharmacokinetics of tolterodine and 5-HM were similar in the presence and absence of warfarin. There were no safety concerns. These findings indicate that co-administration of tolterodine and warfarin is safe and well tolerated, with no clinically significant pharmacodynamic or kinetic interaction in healthy volunteers. PMID- 12572530 TI - Transport of anionic drugs across the basolateral membrane of proximal S2 segments of the rabbit kidney. Inverse relationship between the affinity to the p aminohippurate transport system and the transport rate. AB - The renal p-aminohippurate (PAH) transport system, which resides in the S2 segments of proximal tubules, is a main tubular secretory system for drugs. Previous studies have shown that the hydrophobicity of drugs is positively correlated with their affinity for the basolateral PAH transporter. Affinity was deduced from inhibition of tubular 3H-PAH uptake. However, up to now, no studies are available which tested the relationship between hydrophobicity and transport rate of drugs. Therefore, in the present study, the basolateral transport rates of several drugs (probenecid, CAS 57-66-9, furosemide, CAS 54-31-9, bumetanide, CAS 28395-03-1, mefruside, CAS 7195-27-9) which are substrates of the PAH transporter and which differ markedly regarding their hydrophobic properties (log P values ranging from 1-3) were determined on non-perfused proximal S2 segments microdissected from rabbit kidneys without the use of enzymatic solutions. To measure the transport rate advantage of the finding was taken that the PAH transporter in the basolateral membrane of proximal S2 segments acts as a countertransporter which exchanges PAH with dicarboxylates (glutarate or alpha keto-glutarate). Hence, the stimulatory effect of a drug on the 14C-glutarate efflux rate of 14C-glutarate preloaded S2 segments is a measure of its translocation across the basolateral cell membrane. The results revealed that with increasing hydrophobicity (mefruside < furosemide < bumetanide < probenecid) the stimulating effect of the drugs on 14C-glutarate efflux decreased indicating that the compounds are more slowly translocated although their potency to inhibit 3H-PAH uptake increased. Thus, an inverse relationship between hydrophobicity and hence affinity of the drugs for the PAH transporter and transport rate could be established. PMID- 12572531 TI - Synthesis, antibacterial activity, and toxicity of 7-(isoindolin-5-yl)-4 oxoquinoline-3-carboxylic acids. Discovery of the novel des-F(6)-quinolone antibacterial agent garenoxacin (T-3811 or BMS-284756). AB - The palladium-catalyzed cross-coupling reaction of 5-(tributylstannyl)isoindoline and its 1- and 3-methyl derivatives with 6-fluoro or 6-unsubstituted 7-bromo-1 cyclopropyl-8-methoxy (or difluoromethoxy)-4-oxoquinoline-3-carboxylate afforded the corresponding 1-cyclopropyl-7-(5-isoindolinyl)-4-oxoquinoline-3- carboxylic acids: 6-fluoro, 1a-7a and 6-nonfluoro, 1b-7b. The in vitro antibacterial spectra of the newly synthesized quinolones were mostly characterized by excellent Gram positive activity against Staphylococcus aureus and Streptococcus pneumoniae including quinolone-resistant strains, and also by significant Gram-negative activity comparable to 7-(1-piperazinyl)fluoroquinolones. Comparative examinations of the in vitro antibacterial profiles and the in vivo toxicity in terms of intravenous lethality, micronuclei-inducing potential and convulsive activity provided 6-nonfluorinated 1-cyclopropyl-8-(difluoromethoxy)-7-(1 methylisoindolin-5-yl)-4- oxoquinoline-3-carboxylic acid [(+/-)-5b] as the candidate for evaluation of the stereoisomers. The enantiomers (R)-5b and (S)-5b were synthesized via the Suzuki coupling reaction of (R)- and (S)-1-methyl derivatives of 2-(triphenylmethyl)isoindolin-5-boronic acid with the corresponding 7-bromo-8-(difluoromethoxy)-4- oxoquinoline-3-carboxylate. The (R) 5b stereoisomer proved to be 2- to 4-fold more active than the (S)-5b stereoisomer against the organisms tested, with the exception of an equal potency observed with S. pneumoniae IID553 and Haemophilus influenzae ATCC49247. A noticeable in vitro antibacterial profile of (R)-5b was that it is 16- and 64 fold more active than levofloxacin (CAS 100986-85-4) and ciprofloxacin (CAS 86393 32-0), respectively, against Mycoplasma pneumoniae IID813 (MIC of 0.0313 microgram/ml), and 4-fold more active than ciprofloxacin and levofloxacin against Mycobacterium tuberculosis M-4 (MIC of 0.0313 microgram/ml). Additional studies indicate that (R)-5b (T-3811, CAS 194804-75-6) exhibits excellent antibacterial activity against a wide range of organisms including anaerobes and common respiratory pathogens, while demonstrating a high selectivity against the mammalian homolog topoisomerases. The methane-sulfonate of (R)-5b (T-3811ME, CAS 223652-90-2) is now undergoing clinical testings. PMID- 12572532 TI - Hematopoietic responses under protracted exposures to low daily dose gamma irradiation. AB - In attempting to evaluate the possible health consequences of chronic ionizing radiation exposure during extended space travel (e.g., Mars Mission), ground based experimental studies of the clinical and pathological responses of canines under low daily doses of 60Co gamma irradiation (0.3-26.3 cGy d-1) have been examined. Specific reference was given to responses of the blood forming system. Results suggest that the daily dose rate of 7.5 cGy d-1 represents a threshold below which the hematopoietic system can retain either partial or full trilineal cell-producing capacity (erythropoiesis, myelopoiesis, and megakaryopoiesis) for extended periods of exposure (>1 yr). Trilineal capacity was fully retained for several years of exposure at the lowest dose-rate tested (0.3 cGy d-1) but was completely lost within several hundred days at the highest dose-rate (26.3 cGy d 1). Retention of hematopoietic capacity under chronic exposure has been demonstrated to be mediated by hematopoietic progenitors with acquired radioresistance and repair functions, altered cytogenetics, and cell-cycle characteristics. Radiological, biological, and temporal parameters responsible for these vital acquisitions by hematopoietic progenitors have been partially characterized. These parameters, along with threshold responses, are described and discussed in relation to potential health risks of the space traveler under chronic stress of low-dose irradiation. PMID- 12572533 TI - Impact factor: a valid measure of journal quality? AB - OBJECTIVES: Impact factor, an index based on the frequency with which a journal's articles are cited in scientific publications, is a putative marker of journal quality. However, empiric studies on impact factor's validity as an indicator of quality are lacking. The authors assessed the validity of impact factor as a measure of quality for general medical journals by testing its association with journal quality as rated by clinical practitioners and researchers. METHODS: We surveyed physicians specializing in internal medicine in the United States, randomly sampled from the American Medical Association's Physician Masterfile (practitioner group, n = 113) and from a list of graduates from a national postdoctoral training program in clinical and health services research (research group, n = 151). Respondents rated the quality of nine general medical journals, and we assessed the correlation between these ratings and the journals' impact factors. RESULTS: The correlation between impact factor and physicians' ratings of journal quality was strong (r2 = 0.82, P = 0.001). The correlation was higher for the research group (r2 = 0.83, P = 0.001) than for the practitioner group (r2 = 0.62, P = 0.01). CONCLUSIONS: Impact factor may be a reasonable indicator of quality for general medical journals. PMID- 12572534 TI - [Allozyme-based genetic variation within an unstable "population" of Chinese Oncomelania hupensis (Gastropoda: Rissoacea: Pomatiopsidae)]. AB - OBJECTIVE: To answer the following questions: 1. For Oncomelania snails collected two years apart from the same locality, has there been genetic divergence? 2. How much experimental error has there been in studying subsets of these populations? 3. As this is an unstable population, what has the net effect been on Hardy Weinberg equilibrium (HWe)? METHODS: Allozymes were studied using horizontal starch gel electrophoresis. Data collected from numbers of experiments were compiled. Data from each collection were divided into two equal subsets based on chronology of the experiments. Thirty-four loci were studied using 72 to 180 snails per subset. RESULTS: The mean number of alleles per locus ranged from 1.5 1.9. With each consecutive subset, the % polymorphic loci dropped from 38.2 to 17.6. The mean heterozygosity was very low: 0.033 to 0.049 and not significantly different from Hardy-Weinberg expectations. Ten loci and 11 alleles exclusive to the first group were eliminated from the overall study reducing the number of polymorphic loci from 19 to 10. There were significant departures from HWe at five loci having a substantial number of individuals for each allele. Nei's and Wright's D were 0.003 +/- 0.001 and 0.054 +/- 0.006 respectively. CONCLUSION: 1. There were significant errors seen primarily in the results scored in the earliest experiments. 2. These earlier errors involving scoring difficult to resolve loci, and interpretation of rare alleles that were not found in later experiment had no significant effect on overall genetic distance. 3. The use of Wright's D for closely related populations is explained. Results with Nei's D indicated no significant difference among the four subunits; Wright's D yielded significant difference between the collections made two years apart, attributed to the annual flooding of the Yangtze River mixing snails from different localities. 4. Major polymorphic loci were not in Hwe as predicted using the unstable population model. 5. One must study 25 or more individuals to find relatively rare alleles and study population genetics. PMID- 12572535 TI - Allozyme-based genetic variation within an unstable "population" of Chinese Oncomelania hupensis (Gastropoda: Rissoacea: Pomatiopsidae) (continued). PMID- 12572536 TI - Creating the right chemistry. PMID- 12572537 TI - Facing our demons. PMID- 12572538 TI - New drug approvals for 2002. PMID- 12572540 TI - Bibliography. Current world literature. Ageing: biology and nutrition. PMID- 12572539 TI - Fresh from the pipeline. Ezetimibe. PMID- 12572541 TI - Bibliography. Current world literature. Protein and amino acid metabolism. PMID- 12572543 TI - Radiofrequency ablation: a good regional therapy for liver cancer. PMID- 12572544 TI - Unlock the door to clearer communication. PMID- 12572545 TI - Safety of some surgeries has not improved, study says. PMID- 12572546 TI - Older women with severe asthma are not making the best use of recommended medications. PMID- 12572547 TI - Reviews of hormone replacement therapy evidence support findings in recently halted trial. PMID- 12572548 TI - Care givers burdened and lack support from the healthcare system. PMID- 12572549 TI - [Thomas Eakins: Gross' clinic, 1875]. PMID- 12572550 TI - Bibliography of toxinology. PMID- 12572551 TI - Re: McCabe, Susan. The nature of psychiatric nursing: the intersection of paradigm, evolution and history; Archives of Psychiatric Nursing: April, 2002. PMID- 12572552 TI - [Hashish]. PMID- 12572553 TI - [Migration of pharmacists in Belgium from the 13th through 18th centuries]. AB - The municipal records are important sources for the migration and mobility of the apothecaries from the 13th to the 18th century. They prove that immigration from Italy and France, formerly regarded as the first factor for the presence in our country of pharmacists, was minimal. Only two Italian and two French pharmacists from 1377 unto 1500 could be traced. The second thesis concerning the evolution of the profession, the so called mercer theory, is on the other hand very important. Mobility is here a really substantial component because of the apprentice years. The 18th century represents a growing settlement in small locations. The emigration of the 16th century were occasioned by religious and political factors. PMID- 12572555 TI - John Calvin and Michael Servetus: tragic clash of reformer and radical physician. PMID- 12572554 TI - Clara Louise Maass - nurse (1876-1901). PMID- 12572556 TI - [The encampment of Lazear and the centenary of the confirmation of the Finlay doctrine]. PMID- 12572557 TI - The substrate translocation channel of the proteasome. PMID- 12572558 TI - Philosophy of medicine according to Ayurveda. AB - The modern, western system of medicine researches only a small group of people, it is understood that over half of the population of the world has no share in it. In India the overwhelming majority of people receive medical service from traditional practitioners, who treat their patients according to the principles of Ayurveda. Also in Sri Lanka, Bangladesh and Nepal a vast majority of the masses has been looking up to Ayurveda in its pure form or some of its variations in some form or other, for relief of various ailments. This is how Ayurveda has come to attract the attention of WHO. PMID- 12572559 TI - Historical review of Vicharchika as per Ayurvedic classics. AB - Vicharchika which is one of the common skin diseases, is most prevalent among masses whether rural or urban. A critical peep with regard to this into the various Ayurvedic Classics reveals that, the various kushtha rogas (including Vicharchika) were known to ancients of remote antiquity. Though all Ayurvedic classics have described eighteen varieties of Kushthas dividing into the two main groups viv. Mahakushthas and Kshudra Kushthas, but they differ in nomenclature, order, description, doshas involved and prognosis etc. PMID- 12572560 TI - Polypharmacy and adverse drug-related events. PMID- 12572561 TI - Akathisia can be reduced by lowering the dose of D2 receptor antagonists. PMID- 12572562 TI - The assessment of the amount of fat in the human body from measurements of skinfold thickness. 1967. PMID- 12572563 TI - Human factors in emergency care. PMID- 12572564 TI - Increased risk of intimate partner homicide among California women who purchase handguns. PMID- 12572565 TI - West Nile encephalitis presenting as a stroke. PMID- 12572566 TI - Proceedings of the conference on antibiotic use in animal agriculture. Urbana, Illinois, USA. October 16-17, 2000. PMID- 12572567 TI - Shmuel Shaltiel. PMID- 12572568 TI - Bibliography. Current world literature. Gynecologic oncology and pathology. PMID- 12572569 TI - The ethics of health care: an assessment in Germany and the United States. Proceedings of a conference at the Columbus School of Law at Catholic University. Washington, D.C., USA. October 4, 2001. PMID- 12572570 TI - Glimpses of Islamic medicine. AB - The fall of the Roman Empire during the fifth century A.D. Ushered in the beginning of the Dark Ages. After this, in Europe further progress of Greco-Roman medicine originated from Hippocrates was halted. The ideas about medicine and hygiene were kept alive in monasteries only. The Arabs made advances in medicine at a time when the rest of Europe was in the Dark Ages. Islamic system or the rulers of the day actively encouraged scholarship and growth of knowledge. The Islamic gift of the day to the world of medicine was simply unique. PMID- 12572571 TI - Todaramalla. AB - Todaramalla was a famous physician, scholar, statesman, Finance and Revenue Minister in the court of Akbar. One of the oldest commentary namely 'Manojna' was written by him on Ashtanga Hridaya. He was patriot of Ayurveda and author of 23 series of works in the form of an encyclopedia called 'Todarananda'. Ayurveda saukhyam is one among the Todarananda. The biography of Todaramalla has been presented here. PMID- 12572572 TI - Hormones throughout history. AB - Since remote ages man believed that, the different parts of his body and that of the animals, contained therapeutical elements, with effective powers like magic, and that he can fortify his own personality by consuming some parts of the bodies of human beings or animals. He also believed that his organic diseases were due to deficiency in these organs concerning ailments, so he treated them with similar organs of different animals. PMID- 12572573 TI - Promotive and prophylactic health aspects based on Islamic teachings. AB - Health is basic need of human being and everybody intends to remain healthy. As far as Islam is concerned, its faith is not merely upon spirit, but also it has given proper place to the physician health. It is a fact that the revaluation in the preserving and restoring health of psychic as well as physical is a gift of Islam. Clearcut guidelines have been given not only for correction of spiritual life, but promotive health aspects have also been prescribed at length. In this way the glimpses of promotive and prophylactic health aspects based on Islamic teachings have been given in this article. PMID- 12572574 TI - A rare report of Nizam Ayurvedic Mobile Clinic. AB - This article is a brief account of a rare Urdu booklet entitled 'Report Nizam Ayurvedic Safari Dawakhana'. It is a report of a mobile clinic sent by Nizam VII of Hyderabad to Kumbh Mela at Allahabad on a religious occasion in the year 1942, which was printed in the same year. This report sheds light on the history of the sacred place Prayag, its importance and the activities of the mobile clinic with the remarks of eminent personalities who visited the clinic on the occasion. PMID- 12572575 TI - Hyperbaric oxygen for acute carbon monoxide poisoning. PMID- 12572576 TI - Hyperbaric oxygen for acute carbon monoxide poisoning. PMID- 12572577 TI - Hyperbaric oxygen for acute carbon monoxide poisoning. PMID- 12572578 TI - Hyperbaric oxygen for acute carbon monoxide poisoning. PMID- 12572579 TI - [Analysis and correction of leg deformities. 1: Analysis]. PMID- 12572580 TI - Aspirin, clopidogrel, or both for secondary prevention of coronary disease. PMID- 12572581 TI - Definition of Nidana Panchaka (first chapter of Madhavanidana). AB - The reputation of a physician depends more upon his skill in diagnosis and prognosis. Hence very much importance is given in Ayurveda for the diagnosis of diseases from a consideration of their etiology, symptoms and prognosis, with an aim to arrive at a correct diagnosis before starting any treatment. Madhavakara has brought to perfection the branch of pathology in his work "Madhava Nidana", which represents an attempt towards the revival of the etiopathogenesis, clinical pathology and diagnosis of a large number of diseases together at one place from classical sources. PMID- 12572582 TI - Aspirin, clopidogrel, or both for secondary prevention of coronary disease. PMID- 12572583 TI - Aspirin, clopidogrel, or both for secondary prevention of coronary disease. PMID- 12572584 TI - Aspirin, clopidogrel, or both for secondary prevention of coronary disease. PMID- 12572585 TI - ECVAM's activities in validating alternative tests for skin corrosion and irritation. AB - ECVAM has funded and managed validation studies on in vitro tests for skin corrosion, resulting in the validities of four in vitro tests being endorsed by the ECVAM Scientific Advisory Committee: the rat skin transcutaneous electrical resistance (TER) assay, two tests based on the use of commercial reconstituted human skin equivalents, EPISKIN and EpiDerm, and another commercially-produced test, CORROSITEX. In the European Union (EU), a new test method on skin corrosion (B.40), incorporating the rat skin TER and human skin model assays, was included in Annex V of Directive 67/548/EEC in mid-2000, thereby making the use of in vitro alternatives for skin corrosion testing of chemicals mandatory in the EU. At the recommendation of its Skin Irritation Task Force, ECVAM has funded prevalidation studies on five in vitro tests for acute skin irritation: EpiDerm, EPISKIN, PREDISKIN, the pig-ear test, and the mouse-skin integrity function test (SIFT). However, none of the tests met the criteria (set by the Management Team for the studies) for inclusion in a large-scale formal validation study. Thus, to date, there are no validated in vitro tests for predicting the dermal irritancy of chemicals. Following further work on the EPISKIN, EpiDerm and SIFT test protocols and/or prediction models after the completion of the prevalidation studies, it appears that the modified tests could meet the performance criteria defined for progression to a validation study. This will now be assessed independently by the ECVAM Skin Irritation Task Force, with the objective of taking a decision before the end of 2002 on whether to conduct a formal validation study. PMID- 12572586 TI - Dhanwantari: the God of Hindu medicine. AB - The original conception of Ayurveda in its entirety is essentially linked to Dhanwantari who is considered as God of Hindu Medicine. Dhanwantari is considered a mythical deity born with ambrosia in one hand and Ayurveda on the other at the end of the churning of milk ocean. He reincarnated himself in the Chandra dynasty. He was born to King Dhanwa, learnt Ayurveda from Bharadwaja. His great grandson Divodasa was also known as Dhanwantari, but was specialised only in surgical branch of Ayurveda. Sushruta, is said to have learnt the art of science of surgery from Divodasa Dhanwantara. PMID- 12572587 TI - Current awareness in prenatal diagnosis. PMID- 12572588 TI - Perinatal health--a global perspective. PMID- 12572589 TI - The earliest cells: who are they? PMID- 12572590 TI - Leptin in pregnancy. AB - Leptin is a 16-kDa polypeptide, encoded in humans by the LEP gene. This protein is probably involved in the regulation of ovarian function, oocyte maturation and embryo development, and in the implantation process. During pregnancy, leptin is produced in the placenta; its values increase in the maternal blood particularly in the second trimester and decline postpartum. Leptin levels may be abnormally high in pregnancies complicated by conditions such as diabetes mellitus and pre eclampsia. The role of leptin in fetal physiology has not been clarified, although cord blood values correlate significantly with birth weight. It is expected that further research will provide new insight into the physiological importance of leptin in the pregnant woman and fetus. PMID- 12572591 TI - Protein kinase C stimulates release of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 by human decidual cells. AB - OBJECTIVE: Increased concentrations of amniotic fluid matrix metalloproteinase (MMP)-9 and tissue inhibitor of metalloproteinase (TIMP)-1 have been observed in the context of premature rupture of membranes (PROM) and microbial invasion of the amniotic cavity. However, the source of the stimuli that contribute to the accumulation of these proteins in amniotic fluid remains to be identified. The present study was conducted to investigate MMP-2, MMP-9 and TIMP-1 secretion by decidual cells in response to activated protein kinase C (PKC). METHODS: Decidual cells were isolated from term placentae, grown to confluence and incubated with control media or 10(-11) to 10(-8) mol/l concentrations of phorbol 12-myristate 13-acetate (PMA). Concentrations of MMP-2, MMP-9 and TIMP-1 in the culture supernatant were determined using sensitive and specific immunoassays. Substrate zymography was conducted to confirm MMP-9 assays. RESULTS: PMA induced a concentration-dependent stimulation of release of MMP-9 (control vs. PMA l0(-9) and 10(-8) mol/l; p < 0.01) and TIMP-1 (control vs. PMA 10(-9) and 10(-8) mol/l; p < 0.001), but not MMP-2. A direct positive correlation was observed between MMP 9 and TIMP-1 release (r = 0.645; p < 0.001). Substrate zymography confirmed increased release of MMP-9 in response to PMA (control vs. PMA 10(-8) and PMA 10( 7) mol/l; p < 0.01). CONCLUSIONS: Activation of PKC within the decidua will result in enhanced MMP-9 release, which upon activation could contribute to degradation of matrices within fetal membranes leading to PROM. PMID- 12572592 TI - Neutrophil elastase and secretory leukocyte protease inhibitor in prelabor rupture of membranes, parturition and intra-amniotic infection. AB - OBJECTIVE: Neutrophil elastase (NE), a multifunctional serine protease stored in azurophilic granules of mature neutrophils, is capable of intracellular degradation of proteins during phagocytosis and extracellular degradation of connective tissue during an inflammatory process. Secretory leukocyte protease inhibitor (SLPI) is a natural NE inhibitor present in amniotic fluid, fetal membranes and cervical mucus. An imbalance between NE and SLPI has been implicated as a mechanism of abnormal tissue destruction in chronic inflammatory diseases. The purpose of this study was to determine if parturition, premature rupture of the membranes (PROM) and microbial invasion of the amniotic cavity are associated with changes in amniotic fluid concentrations of NE and SLPI. STUDY DESIGN: Amniotic fluid was retrieved by amniocentesis from 380 patients in the following groups: (1) preterm labor and intact membranes without microbial invasion of the amniotic cavity who delivered at term (n = 13) or prematurely (n = 26), and preterm labor with microbial invasion of the amniotic cavity (n = 9); (2) preterm PROM with (n = 34) and without (n = 51) microbial invasion of the amniotic cavity; and (3) term gestation without microbial invasion of the amniotic cavity with intact membranes not in labor (n = 63), in labor (n = 158), and with rupture of membranes not in labor (n = 26). Microbial invasion of the amniotic cavity was determined by a positive amniotic fluid culture for micro organisms including aerobic, anaerobic and Mycoplasma species. NE and SLPI amniotic fluid levels were determined by highly specific and sensitive immunoassays. RESULTS: Preterm PROM was associated with a significant increase in the amniotic fluid concentration of NE. Microbial invasion of the amniotic cavity was associated with a significant increase in the amniotic fluid concentration of NE in women with preterm labor and intact membranes, as well as in women with preterm PROM. Term and preterm parturition was associated with a significant increase in the amniotic fluid concentration of NE. In the absence of microbial invasion of the amniotic cavity, preterm and term PROM were associated with a significant reduction in the amniotic fluid concentration of SLPI. CONCLUSION: Preterm PROM, microbial invasion of the amniotic cavity, and parturition at term and preterm are associated with a significant increase in the amniotic fluid concentration of NE. PROM is associated with a reduced amniotic fluid concentration of SLPI. PMID- 12572593 TI - Comparison of fetal growth estimates based on birth weight and ultrasound references. AB - OBJECTIVE: To compare three different methods for modeling fetal weight gain during the third trimester of pregnancy. METHODS: Ultrasound and live birth weight data were used to construct three models for defining fetal growth during the third trimester: longitudinal ultrasound estimates of fetal weight obtained serially at 3-4 week intervals in 50 uncomplicated, well-dated pregnancies between 19 and 40 weeks' gestation; cross-sectional ultrasound estimates of fetal weight obtained from 2018 ultrasound examinations of singleton, non-anomalous fetuses between 24 and 39 weeks' gestation; and cross-sectional birth weight data obtained from 9553 live singleton, non-anomalous neonates between 24 and 43 completed weeks. Analysis was performed by pairwise partial f test to compare regression curves and zeta test for comparison of mean weekly weight gain. A value of p < 0.05 was accepted for significance. RESULTS: Derived regression lines depicting fetal size across gestation were significantly different from each other (f tests, p < 0.05). Estimates of mean fetal weight were significantly different between the three different models at specific gestational ages. Significant weekly variations in fetal weight gain were observed within the raw cross-sectional data sets, both for ultrasound-estimated fetal weight (range 91 278 g/week) and birth weight (65-309 g/week). CONCLUSIONS: Each of the methods used to model normal fetal weight gain in the third trimester defined a distinct pattern of fetal growth. Normal fetal growth, defined longitudinally, was most closely matched by a combination of cross-sectional ultrasound-derived estimated fetal weight in preterm gestation below 34 weeks' gestation and live birth weight at or beyond 34 weeks. PMID- 12572594 TI - Late-onset fetal cardiac decelerations associated with fetal breathing movements. AB - BACKGROUND: While late decelerations are regarded as signs of fetal hypoxemia, fetal breathing movements (FBM) associated with late decelerations invariably have normal outcomes. Could late decelerations sometimes represent FBM? MATERIALS AND METHODS: Six patients between 37 and 42 weeks' gestation with 'late decelerations' associated with FBM (by ultrasound or tocodynamometer) during ante or intrapartum monitoring were evaluated. Three were at high risk (diabetes, postdates, intrauterine growth restriction) and three were at low-risk. RESULTS: 'Late decelerations' arose in previously reassuring tracings. Oxygen or positional change had no effect. The decelerations were variable in length and shallow, and contained increased variability. Normal baseline rate and variability were maintained after the deceleration. Neonatal outcomes were normal. CONCLUSION: 'Late decelerations' as described are associated with normal outcome and may represent FBM. This understanding may reduce unnecessary interventions. PMID- 12572595 TI - ST analysis of the fetal electrocardiogram during labor: Nordic observational multicenter study. AB - OBJECTIVE: To assess the diagnostic power of cardiotocography (CTG) plus the ST interval of the electrocardiogram (ECG) clinical guidelines with combined fetal heart rate and ST waveform analysis of the fetal ECG recorded during labor, to identify an adverse labor outcome (neonatal neurological symptoms and/or metabolic acidosis). STUDY DESIGN: An observational, multicenter study was undertaken in 12 Nordic labor wards. A total of 573 women in labor were monitored using a prototype of the STAN S 21 recorder with fetal ECG data and computerized ST analysis. RESULTS: Fifteen cases of intrapartum fetal hypoxia identified from neurological neonatal symptoms and/or cord artery pH < 7.05 with base deficit in extracellular fluid > 12.0 mmol/l were recorded. All these cases were identified by CTG + ST clinical guidelines. Five developed neonatal symptoms and had ECG abnormalities during the first stage of labor and, of the remaining ten, eight showed ST changes during active pushing in the second stage. Another eight cases had acidemia only and normal neonatal outcome. Seven of these displayed CTG + ST abnormalities. The high sensitivity of CTG + ST to predict fetal acidosis was associated with a marked increase in positive predictive values compared with conventional CTG. CONCLUSION: The STAN clinical guidelines identify fetuses at risk of intrapartum asphyxia. PMID- 12572596 TI - Factor V Leiden and prothrombin G20210A mutations in pregnancies with adverse outcome. AB - BACKGROUND: Inherited thrombophilia has been associated with obstetric complications through mechanisms that are not yet fully elucidated. The aim of this study was to investigate the relationship between specific obstetric adverse outcomes and factor V Leiden and prothrombin G20210A mutations. METHODS: Forty five women with adverse pregnancy outcome defined as severe pre-eclampsia, abruptio placentae, intrauterine growth restriction and stillbirth, were tested for factor V Leiden and prothrombin G20210A mutations. The control group comprised 100 women with at least one normal pregnancy and no history of thrombosis. RESULTS: Overall, 13 women with one or more of the above-mentioned pregnancy complications (28%) had either thrombophilic mutation, as compared with six in the control group (6%) (p < 0.001, odds ratio (OR) 6.1; 95% confidence interval (CI) 1.9-20). The factor V Leiden mutation was detected in ten of the women with complicated pregnancies (22%) and in four of the controls (4%) (p < 0.001, OR 6.6; 95% CI 1.7-27.2). The prothrombin G20210A mutation was detected in three women in the group with complications (6%) and in two of the controls (2%) (p = 0.17, OR 3.4; 95% CI 0.4-30.5). Compared to controls, the prevalence of the factor V Leiden mutation was significantly higher in the subgroups of severe pre eclampsia, abruptio placentae and fetal growth restriction. The prevalence of the prothrombin G20210A mutation does not appear to be significantly different from that in the controls in any of the groups studied. CONCLUSIONS: Our data suggest that inherited thrombophilia, and specifically the factor V Leiden mutation, may be associated with adverse pregnancy outcome. The role of the prothrombin G20210A mutation remains to be elucidated. PMID- 12572597 TI - Postpartum maternal levels of hemoglobin A1c and cord C-peptide in macrosomic infants of non-diabetic mothers. AB - OBJECTIVE: This study was designed to test the hypothesis that macrosomia in infants born to non-diabetic mothers is associated with an increased incidence of hyperinsulinemia and normal maternal glucose regulation in late pregnancy. METHODS: Twenty mothers and their macrosomic infants were chosen as the study group, and 20 mothers with their appropriate-for-gestational-age infants were chosen as the control group. RESULTS: No difference in postpartum mean hemoglobin A1c levels was observed between the mothers of macrosomic infants and those of control infants. Cord plasma C-peptide levels were significantly higher in macrosomic than in control infants. CONCLUSIONS: This study revealed that macrosomic infants of non-diabetic mothers were significantly more likely to have hyperinsulinemia than were normal-sized infants, and this hyperinsulinemia was not caused by dysregulation in glucose metabolism. PMID- 12572598 TI - Acute fatty liver of pregnancy. AB - Acute fatty liver of pregnancy can produce a great variety of complications. Of these, infections are important. We describe two cases, one of them complicated with uterine sepsis. Both gave evidence of the necessity for multidisciplinary treatment, which reduces the maternal-fetal morbidity and mortality produced by this disease. PMID- 12572599 TI - Incidental tubo-ovarian abscess at abdominal delivery: a case report. AB - Tubo-ovarian abscess in the third trimester of pregnancy is extremely rare. In this report, an unusual case with asymptomatic tubo-ovarian abscess, diagnosed incidentally during Cesarean section performed for an obstetric indication, is presented. Unlike other reported cases, no signs or symptoms attributable to pelvic abscess throughout the pregnancy were observed in our patient. To our knowledge, this is the first report of such a case in the literature. PMID- 12572600 TI - Brenner tumor of the ovary with extensive stromal luteinization presenting in pregnancy: report of a case and review of the literature. AB - Brenner tumor in pregnancy is rare, with only two reported cases in association with intrauterine pregnancy. A 30-year-old woman, G1P0, was found to have an adnexal mass at 15 weeks of gestation. The mass was resected at Cesarean section and found to be a Brenner tumor with areas of stromal luteinization. Brenner tumor should be considered in the differential diagnosis of adnexal masses presenting during pregnancy. The stromal luteinization was probably related to the hormonal environment of the pregnancy. PMID- 12572601 TI - Complex history of a chromosomal paralogy region: insights from amphioxus aromatic amino acid hydroxylase genes and insulin-related genes. AB - Aromatic amino acid hydroxylase (AAAH) genes and insulin-like genes form part of an extensive paralogy region shared by human chromosomes 11 and 12, thought to have arisen by tetraploidy in early vertebrate evolution. Cloning of a complementary DNA (cDNA) for an amphioxus (Branchiostoma floridae) hydroxylase gene (AmphiPAH) allowed us to investigate the ancestry of the human chromosome 11/12 paralogy region. Molecular phylogenetic evidence reveals that AmphiPAH is orthologous to vertebrate phenylalanine (PAH) genes; the implication is that all three vertebrate AAAH genes arose early in metazoan evolution, predating vertebrates. In contrast, our phylogenetic analysis of amphioxus and vertebrate insulin-related gene sequences is consistent with duplication of these genes during early chordate ancestry. The conclusion is that two tightly linked gene families on human chromosomes 11 and 12 were not duplicated coincidentally. We rationalize this paradox by invoking gene loss in the AAAH gene family and conclude that paralogous genes shared by paralogous chromosomes need not have identical evolutionary histories. PMID- 12572602 TI - Nucleotide substitution models and estimation of phylogeny. AB - The nucleotide substitution matrix inferred from avian data sets using cytochrome b differs considerably from the models commonly used in phylogenetic analyses. To analyze the possible effects of this particular pattern of change in phylogeny estimation we performed a computer simulation in which we started with a real sequence and used the inferred model of change to produce a tree of 10 species. Maximum parsimony (MP), maximum likelihood (ML), and various distance methods were then used to recover the topology and the branch lengths. We used two kinds of data with varying levels of variation. In addition, we tested with the removal of third positions and different weighting schemes. At low levels of variation, MP was outstanding in recovering the topology (90% correct), while unweighted pair-group method, arithmetic average (UPGMA), regardless of distances used, was poor (40%). At the higher level, most methods had a chance of around 40%-58% of finding the true tree. However, in most cases, the trees found were only slightly wrong, with only one or a few branches misplaced. On the other hand, the use of a "wrong" model had serious effects on the estimation of branch lengths (distances). Although precision was high, accuracy was poor with most methods, giving branch lengths that were biased downward. When seeded with the true distance matrix, Fitch and NJ always found the true tree, while UPGMA frequently failed to do so. The effect of removing third positions was dramatic at low levels of variation, because only one MP program was able to find a true tree at all, albeit rarely, while none of the others ever did so. At higher levels, the situation was better, but still much worse than with the whole data set. PMID- 12572603 TI - Evolutionary relationships among the members of an ancient class of non-LTR retrotransposons found in the nematode Caenorhabditis elegans. AB - We took advantage of the massive amount of sequence information generated by the Caenorhabditis elegans genome project to perform a comprehensive analysis of a group of over 100 related sequences that has allowed us to describe two new C. elegans non-LTR retrotransposons. We named them Sam and Frodo. We also determined that several highly divergent subfamilies of both elements exist in C. elegans. It is likely that several master copies have been active at the same time in C. elegans, although only a few copies of both Sam and Frodo have characteristics that are compatible with them being active today. We discuss whether it is more appropriate under these circumstances to define only 2 elements corresponding to the most divergent groups of sequences or up to 16, considering each subfamily a different element. The C. elegans elements are related to other previously described non-LTR retrotransposons (CR1, found in different vertebrates; SR1, from the trematode Schistosoma; Q and T1, from the mosquito Anopheles). All of these elements, according to the analysis of their reverse transcriptases, form a monophyletic cluster that we call the "T1/CR1 subgroup." Elements of this subgroup are thus ancient components of the genome of animal species. However, we discuss the possibility that these elements may occasionally be horizontally transmitted. PMID- 12572604 TI - Microevolutionary divergence pattern of the segmentation gene hunchback in Drosophila. AB - To study the microevolutionary processes shaping the evolution of the segmentation gene hunchback (hb) from Drosophila melanogaster, we cloned and sequenced the gene from 12 isofemale lines representing wild-type populations of D. melanogaster, as well as from the closely related species Drosophila sechellia, Drosophila orena, and Drosophila yakuba. We find a relatively low degree of sequence variation in D. melanogaster (theta = 0.0017), which is, however, consistent with its chromosomal location in a region of low recombination. Tests of neutrality do not reject a neutral-evolution model for the whole region. However, pairwise tests with different subregions indicate that there is a relative excess of polymorphic sites in the leader and the intron. Codon usage pattern analysis shows a particularly biased codon usage in the highly conserved regions, which is in line with the hypothesis that selection on translational accuracy is the driving force behind such a bias. A comparison of the expression pattern of hb in different sibling species of D. melanogaster reveals some regulatory changes in D. yakuba, which could be interpreted as changes in the timing of secondary expression domains. PMID- 12572605 TI - Evolution of RPS4Y. AB - Sequence variation within RPS4Y, a ribosomal protein gene located in the nonpseudoautosomal region of the Y chromosome, was used to elucidate the origin of this gene in primates. Complete coding and additional flanking sequences (949 bp) of the RPS4Y locus were determined in four nonhuman primate species. Phylogenetic reconstruction of RPS4 sequence evolution supports the monophyly of mammalian RPS4 and RPS4Y. Molecular evolutionary rate estimation reveals significantly elevated rates of DNA and protein evolution in RPS4Y compared with its X-chromosome homologs. These rates enable us to estimate the timing of the transposition of RPS4X to the Y chromosome (95% confidence interval, 32 MYA-74 MYA), and this estimate was verified by Southern hybridization analysis of prosimian and simian genomic DNA. These data support a transposition event of ancestral primate RPS4X to the Y chromosome prior to the divergence of Prosimii. PMID- 12572606 TI - Intra- and interspecific DNA variation and codon bias of the alcohol dehydrogenase (Adh) locus in Arabis and Arabidopsis species. AB - Sequence variation at the alcohol dehydrogenase (Adh) locus was analyzed for six species each of the genera Arabis and Arabidopsis. Phylogenetic analysis showed that investigated species were grouped into three clusters, and the generic classification did not correspond to the clusterings. The results indicated that the genera could not be distinguished on the basis of the Adh variation. A significant difference in the ratio of silent to replacement sites was detected by MK test in two comparisons, with Arabidopsis thaliana polymorphism due to excess silent divergence. Silent changes were predominant in the evolution of the Adh locus in Arabis and Arabidopsis. To infer evolutionary significance of silent substitutions, codon bias was studied. The degree of codon bias of the Adh region was relatively constant over Arabis and Arabidopsis species. "Preferred" codons of A. thaliana were determined. No evidence of natural selection on codon change was detected in the Adh regions of A. thaliana and Arabis gemmifera. PMID- 12572607 TI - The origin and evolution of variable-region helices in V4 and V7 of the small subunit ribosomal RNA of branchiopod crustaceans. AB - We sequenced the V4 and V7 regions of the small-subunit ribosomal RNA (SSU rRNA) from 38 species of branchiopod crustaceans (e.g., Artemia, Daphnia, Triops) representing all eight extant orders. Ancestral large-bodied taxa in the orders Anostraca, Notostraca, Laevicaudata, and Spinicaudata (limnadiids and cyzicids) possess the typical secondary structure in these regions, whereas the spinicaudatan Cyclestheria and all of the cladocerans (Anomopoda, Ctenopoda, Onychopoda, and Haplopoda) possess three unique helices. Although the lengths and primary sequences of the distal ends of these helices are extremely variable, their locations, secondary structures, and primary sequences at the proximal end are conserved, indicating that they are homologous. This evidence supports the classical view that Cladocera is a monophyletic group and that the cyclestheriids are transitional between spinicaudatans and cladocerans. The single origin and persistence since the Permian of the unique cladoceran helices suggests that births and deaths of variable region helices have been rare. The broad range of sequence divergences observed among the cladoceran helices permitted us to make inferences about their evolution. Although their proximal ends are very GC biased, there is a significant negative correlation between length and GC content due to an increasing proportion of U at their distal ends. Slippage-like processes occurring at unpaired nucleotides or bulges, which are very U-biased, are associated with both helix origin and runaway length expansion. The overall GC contents and lengths of V4 and V7 are highly correlated. More surprisingly, the lengths of these SSU rRNA variable regions are also highly correlated with the length of the large-subunit rRNA expansion segment, D2, indicating that mechanisms affecting length variation do so both across single genes and across genes in the rRNA gene family. PMID- 12572608 TI - Divergent structures of Caenorhabditis elegans cytochrome P450 genes suggest the frequent loss and gain of introns during the evolution of nematodes. AB - The Caenorhabditis elegans genome contains more than 60 cytochrome P450 (CYP) genes. The exon-intron organizations of all of the available and potentially active C. elegans CYP genes were inferred by a newly developed program for predicting protein-coding exons based on the alignment of a genomic DNA sequence and a protein profile. From the predicted amino acid sequences, all of the C. elegans CYP genes except one were classified into three groups, which were closely related to the mammalian drug-metabolizing P450 gene families CYP2, CYP3, and CYP4. The gene structures were strikingly divergent within each group; 20, 10, and 5 unique gene organizations were identified among 40, 18, and 5 genes in the CYP2-, CYP3-, and CYP4-related groups, respectively. The degrees of divergence in gene organization were strongly correlated with those in the amino acid sequences of encoding proteins, and the minimum rate of change in an intron insertion site was estimated to be about 90 times less frequent than amino acid substitutions. Parsimonious analyses suggested that frequent loss and gain of introns has occurred during the evolution of CYP genes in each group after the divergence of nematodes, arthropods, and deuterostomia. Few, if any, incidents of intron sliding were evident, and a model that did not allow intron insertions was highly inconsistent with the observations. All of these findings are explained better by the intron-late view than by the intron-early view. PMID- 12572609 TI - SINE cousins: the 3'-end tails of the two oldest and distantly related families of SINEs are descended from the 3' ends of LINEs with the same genealogical origin. AB - AFC short interspersed elements (SINEs) were isolated from cichlids from Madagascar, the New World, and Africa and characterized. A new family of long interspersed elements (LINEs), designated the CiLINE2 family, was also isolated from African cichlids, and its consensus sequence was deduced. Upon aligning all of the consensus sequences, we found that the 3'-tail regions of the AFC SINEs and the CiLINE2 family were very similar, providing another example in which a reverse transcriptase responsible for retroposition of SINEs might be contributed in trans by a LINE. Sequence comparisons showed that CiLINE2 in cichlids was closely related to LINE2 in mammals. Furthermore, we found that the 3'-tail sequence shared by the AFC SINEs and CiLINE2 in cichlids was very similar to the 3'-tail sequence shared by the MIR SINEs and LINE2 in mammals, even though the remaining parts of the AFC SINEs and the MIR SINEs were totally different from each other. Thus, the present report not only describes a new pair of SINEs and LINEs with the same 3' tail in cichlids, but also provides a new example of the phenomenon whereby the 3' ends of LINEs with the same genealogical origin can be incorporated into the 3'-end tails of different families of SINEs that have been generated independently in two different lineages during evolution. PMID- 12572610 TI - Messinian salinity crisis and the origin of freshwater lifestyle in western Mediterranean gobies. AB - The present paper reports on a molecular study based on 12S rRNA and 16S rRNA mitochondrial genes partly sequenced in 13 species of western Mediterranean gobies, three of which are strictly freshwater-dwelling. A total of 867 bp were aligned and used for the phylogenetic reconstruction. Two major lineages were identified, one clustering the sand gobies in a monophyletic clade. Relationships among taxa based on sequence analysis only partly match those based on morphological criteria, suggesting that the latter are somehow insufficient to correctly establish phylogenetic relationships within this family. The results provide evidence for a multiple independent evolution of the freshwater lifestyle in Knipowitschia and Padogobius lineages. On the basis of the present results, it is uncertain whether the freshwater preference within the genus Padogobius originated twice independently in P. nigricans and P. martensii or only once in their common ancestor. Estimation of the ages of the two major lineages of this group of fish with a molecular clock (in combination with the construction of a linearized tree) suggests that they are much older (at least 40 Myr) than previously thought. Thus, there should be no correlation between their diversification and the Miocene-Pliocene geological events, including the so called Messinian salinity crisis, which occurred about 10 MYA and is believed to have played a role in their evolution. Alternatively, these gobies would have an evolutionary rate at least fourfold faster than those of other vertebrates. PMID- 12572611 TI - A comparative summary of genetic distances in the vertebrates from the mitochondrial cytochrome b gene. AB - Mitochondrial cytochrome b (cytb) is among the most extensively sequenced genes to date across the vertebrates. Here, we employ nearly 2,000 cytb gene sequences from GenBank to calculate and compare levels of genetic distance between sister species, congeneric species, and confamilial genera within and across the major vertebrate taxonomic classes. The results of these analyses parallel and reinforce some of the principal trends in genetic distance estimates previously reported in a summary of the multilocus allozyme literature. In particular, surveyed avian taxa on average show significantly less genetic divergence than do same-rank taxa surveyed in other vertebrate groups, notably amphibians and reptiles. Various biological possibilities and taxonomic "artifacts" are considered that might account for this pattern. Regardless of the explanation, by the yardstick of genetic divergence in this mtDNA gene, as well as genetic distances in allozymes, there is rather poor equivalency of taxonomic rank across some of the vertebrates. PMID- 12572612 TI - Structure and evolution of linalool synthase. AB - Plant terpene synthases constitute a group of evolutionarily related enzymes. Within this group, however, enzymes that employ two different catalytic mechanisms, and their associated unique domains, are known. We investigated the structure of the gene encoding linalool synthase (LIS), an enzyme that uses geranyl pyrophosphate as a substrate and catalyzes the formation of linalool, an acyclic monoterpene found in the floral scents of many plants. Although LIS employs one catalytic mechanism (exemplified by limonene synthase [LMS]), it has sequence motifs indicative of both LMS-type synthases and the terpene synthases employing a different mechanism (exemplified by copalyl diphosphate synthase [CPS]). Here, we report that LIS genes analyzed from several species encode proteins that have overall 40%-96% identity to each other and have 11 introns in identical positions. Only the region encoding roughly the last half of the LIS gene (exons 9-12) has a gene structure similar to that of the LMS-type genes. On the other hand, in the first part of the LIS gene (exons 1-8), LIS gene structure is essentially identical to that found in the first half of the gene encoding CPS. In addition, the level of similarity in the coding information of this region between the LIS and CPS genes is also significant, whereas the second half of the LIS protein is most similar to LMS-type synthases. Thus, LIS appears to be a composite gene which might have evolved from a recombination event between two different types of terpene synthases. The combined evolutionary mechanisms of duplication followed by divergence and/or "domain swapping" may explain the extraordinarily large diversity of proteins found in the plant terpene synthase family. PMID- 12572613 TI - Preponderance of slightly deleterious polymorphism in mitochondrial DNA: nonsynonymous/synonymous rate ratio is much higher within species than between species. AB - We estimated synonymous (dN) and nonsynonymous (dS) substitution rates for protein-coding genes of the mitochondrial genome from two individuals each of the species human, chimpanzee, and gorilla. The genes were analyzed both separately and in a combined data set. Pairwise sequence comparisons suggest that the dN/dS rate ratios are about 5-10 times higher in within-species comparisons than in between-species comparisons. This result is confirmed by a more rigorous likelihood ratio test, which rejected the null hypothesis that the dN/dS rate ratios are identical within and between species. The likelihood models account for the genetic code structure, transition/transversion rate ratio, and codon usage bias and are expected to produce more reliable results than the commonly used contingency test. Separate analyses of different genes show that the dN/dS rate ratios are higher within species than between species for all 13 mitochondrial genes, with the difference being statistically significant for all except three small or slowly evolving genes. Furthermore, in conserved genes, nonsynonymous rates within species tend to be higher than the between-species rates by a greater proportion than in fast-changing genes. Our findings confirm and extend earlier results obtained from smaller data sets and suggest the operation of slightly deleterious mutations throughout the mitochondrial genome in the hominoids. Implications of the results for evolutionary studies and, in particular, for studies of the origin of modern humans, are discussed. PMID- 12572614 TI - Phylogenetic characterization and molecular evolution of bacterial endosymbionts in psyllids (Hemiptera: Sternorrhyncha). AB - Most sternorrhynchan insects harbor endosymbiotic bacteria in specialized cells (bacteriocytes) near the gut which provide essential nutrients for hosts. In lineages investigated so far with molecular methods (aphids, mealybugs, whiteflies), endosymbionts apparently have arisen from independent infections of common host ancestors and co-speciated with their hosts. Some endosymbionts also exhibit putatively negative genetic effects from their symbiotic association. In this study, the identity of endosymbionts in one major sternorrhynchan lineage, psyllids (Psylloidea), was investigated to determine their position in eubacterial phylogeny and their relationship to other sternorrhynchan endosymbionts. Small-subunit ribosomal RNA genes (16S rDNA) from bacteria in three psyllid species (families Psyllidae and Triozidae) were sequenced and incorporated into an alignment including other insect endosymbionts and free living bacteria. In phylogenetic analysis, all sequences were placed within the gamma subdivision of the Proteobacteria. Three sequences, one from each psyllid species, formed a highly supported monophyletic group whose branching order matched the host phylogeny, and also exhibited accelerated rates of evolution and mutational bias toward A and T nucleotides. These attributes, characteristic of primary (P) bacteriocyte-dwelling endosymbionts, suggested that these sequences were from the putative psyllid P endosymbiont. Two other sequences were placed within the gamma-3 subgroup of Proteobacteria and were hypothesized to be secondary endosymbionts. The analysis also suggested a sister relationship between P endosymbionts of psyllids and whiteflies. Thus, a continuous mutualistic association between bacteria and insects may have existed since the common ancestor of psyllids and whiteflies. Calculations using a universal substitution rate in bacteria corrected for endosymbiont rate acceleration support the idea that this common ancestor was also the ancestor of all Sternorrhyncha. Compared with other P endosymbiont lineages, the genetic consequences of intracellular life for some psyllid endosymbionts have been exaggerated, indicating possible differences in population structures of bacteria and/or hosts. PMID- 12572615 TI - Accelerated evolutionary rate in sulfur-oxidizing endosymbiotic bacteria associated with the mode of symbiont transmission. AB - The nearly neutral theory of molecular evolution predicts that the rate of nucleotide substitution should accelerate in small populations at sites under low selective constraint. We examined these predictions with respect to the relative population sizes for three bacterial life histories within chemolithoautotrophic sulfur-oxidizing bacteria: (1) free-living bacteria, (2) environmentally captured symbionts, and (3) maternally transmitted symbionts. Both relative rates of nucleotide substitution and relative ratios of loop, stem, and domain substitutions from 1,165 nt of the small-subunit 16S rDNA were consistent with expectations of the nearly neutral theory. Relative to free-living sulfur oxidizing autotrophic bacteria, the maternally transmitted symbionts have faster substitution rates overall and also in low-constraint domains of 16S rDNA. Nucleotide substitition rates also differ between loop and stem positions. All of these findings are consistent with the predictions that these symbionts have relatively small effective population sizes. In contrast, the rates of nucleotide substitution in environmentally captured symbionts are slower, particularly in high-constraint domains, than in free-living bacteria. PMID- 12572616 TI - The minisatellite MSB1, in the fungus Botrytis cinerea, probably mutates by slippage. AB - A minisatellite was identified in the intron of the ATP synthase of the filamentous fungus Botrytis cinerea, and it was named MSB1. This is the second fungal minisatellite described to date. Its 37-bp repeat unit is AT-rich, and it is found at only one locus in the genome. The introns of 47 isolates of Botrytis species were sequenced. The number of tandem repeats varied only from 5 to 11, but there were many repeat variants. The structure of MSB1 is peculiar: the variants are in the same physical order in all individuals, and this order follows the most parsimonious tree. These original characteristics, together with a total lack of recombination between alleles of the flanking regions, suggest that MSB1 probably mutates by slippage. MSB1 was found in the intron of the ATP synthase of all of the Botrytis species analyzed, but the repeat unit was not found in any other genus examined, including Sclerotinia, which is the genus closest to Botrytis. PMID- 12572617 TI - The evolutionary history of Drosophila buzzatii. XXXIV. The distribution of the retrotransposon Osvaldo in original and colonizing populations. AB - The frequency distribution of the retrotransposon Osvaldo in the haploid genome of Drosophila buzzatii has been studied in five natural populations from the Iberian Peninsula and six natural populations from Argentina. In Iberian populations, Osvaldo insertion sites do not follow a Poisson distribution, most probably due to eight euchromatic sites with high occupancy, found in all populations. The estimated alpha and beta parameters, which measure the relative importance of drift and negative selection in shaping frequency distributions, indicate that drift is the main force acting upon the distribution of Osvaldo in natural populations of D. buzzatii in the Iberian Peninsula. On the other hand, Osvaldo distribution in populations from Argentina is similar to the distribution of elements with low copy numbers, such as those described for Drosophila melanogaster and Drosophila simulans: there are no indications for deviation from a Poisson distribution, there is a low occupancy per insertion site, and genetic drift has no apparent effect on the frequency distribution. We propose that the unusual distribution found in the populations from the Iberian Peninsula is a consequence of the colonization process. Iberian Peninsula populations suffered a genomic redistribution of Osvaldo, most probably after a founder effect. Consequently, certain copies that arrived at high frequencies are showing a high occupancies today, and the mean copy number of Osvaldo is higher in Iberian Peninsula populations than in populations from Argentina. All other copies are the result of recent (after colonization) transposition events. PMID- 12572618 TI - Evolution of genes, evolution of species: the case of aminoacyl-tRNA synthetases. AB - All of the aminoacyl-tRNA synthetase (aaRS) sequences currently available in the data banks have been subjected to a systematic analysis aimed at finding gene duplications, genetic recombinations, and horizontal transfers. Evidence is provided for the occurrence (or probable occurrence) of such phenomena within this class of enzymes. In particular, it is suggested that the monomeric PheRS from the yeast mitochondrion is a chimera of the alpha and beta chains of the standard tetrameric protein. In addition, it is proposed that the dimeric and tetrameric forms of GlyRS are the result of a double and independent acquisition of the same specificity within two different subclasses of aaRS. The phylogenetic reconstructions of the evolutionary histories of the genes encoding aaRS are shown to be extremely diverse. While large segments of the population are consistent with the broad grouping into the three Woesian domains, some phylogenetic reconstructions do not place the Archae and the Eucarya as sister groups but, rather, show a gram-negative bacteria/eukaryote clustering. In addition, many individual genes pose difficulties that preclude any simple evolutionary scheme. Thus, aaRS's are clearly a paradigm of F. Jacob's "odd jobs of evolution" but, on the whole, do not call into question the evolutionary scenario originally proposed by Woese and subsequently refined by others. PMID- 12572619 TI - Pseudogene evolution in Drosophila suggests a high rate of DNA loss. PMID- 12572620 TI - An extra nucleotide is not translated in mitochondrial ND3 of some birds and turtles. PMID- 12572621 TI - Support for a monophyletic lemuriformes: overcoming incongruence between data partitions. PMID- 12572622 TI - The national cholesterol education program adult treatment panel ill guidelines. AB - Coronary heart disease (CHD) remains the leading cause of death in the United States with more than 40% of all deaths each year directly attributed to the disease. Current evidence suggests that early identification and aggressive modification of risk factors offer the most promising approach to reducing the burden of CHD. Dyslipidemia has been identified as the primary risk factor leading to the development of CHD. It is estimated that nearly 65 million Americans require some form of lipid-modification therapy. The National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) set of guidelines released in May 2001 provides physicians with evidence-based recommendations on the classification, diagnosis, and treatment of lipid disorders. New features of the guidelines include a scoring system for calculating CHD risk, as well as the identification of CHD risk equivalents, lower treatment target goals, and an emphasis on conditions conferring a higher risk for CHD, such as the metabolic syndrome. The ATP III emphasis on risk assessment substantially increases the number of patients considered at risk for CHD and will expand the number eligible for lifestyle and drug interventions. This article highlights the new recommendations and reviews the impact of ATP III on osteopathic physicians. PMID- 12572623 TI - Underidentification and undertreatment of dyslipidemia. AB - Despite increased attention placed on the identification and treatment of dyslipidemia, this condition remains undiagnosed and untreated in a significant number of patients. The recently released National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) set of cholesterol management guidelines increases to more than 65 million the number of Americans eligible for lipid-modifying therapy. Recent data, however, suggest that even with the availability of multiple regimens with proven efficacy, as many as 50% of all patients do not have their cholesterol assessed and less than 45% receive lipid modifying therapy. In addition, less than 25% of patients are treated to their NCEP target low-density lipoprotein cholesterol (LDL-C) level. Persistence with therapy is another challenge, as more than 70% of patients fail to maintain their therapy beyond 12 months. If a realistic attempt is to be made to reduce the risk of coronary heart disease (CHD) among Americans, diagnosis of dyslipidemia and treatment to therapeutic targets must be improved. This artide discusses the underdiagnosis and undertreatment of lipid disorders and reviews the role of osteopathic physicians in strategies achieving ATP III LDL-C goals. PMID- 12572624 TI - Combination therapy for dyslipidemia. AB - In the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) guidelines, the emphasis of lipid-lowering therapy is placed on reaching target plasma low-density lipoprotein cholesterol (LDL-C) levels in order to reduce the risk for coronary heart disease (CHD). Although therapeutic lifestyle changes can have a positive effect on LDL-C levels, the ATP III recognizes that a majority of patients with dyslipidemia will also require drug therapy to achieve lipid targets. Currently, only a small percentage of patients, including those with CHD, are reaching goal. Early aggressive use of the effective lipid-lowering agents currently available is critical to achieve target lipid levels in a greater number of patients. Use of drug combinations further enhances the likelihood of achieving target lipid levels. Ideally, the combination of therapeutic modalities used both the endogenous and exogenous pathways of cholesterol synthesis to reduce the amount produced in the body, as well as the amount absorbed from the diet. This article reviews the pharmacotherapeutic effects of combination therapy, summarizes the strengths and weaknesses of current lipid-lowering drug combinations, and identifies the potential impact of the novel cholesterol absorption inhibitor ezetimibe on the LDL-C treatment algorithm. PMID- 12572625 TI - Strategies for optimizing lipid treatment outcomes. AB - The relationship between increased levels of cholesterol and elevated risk for coronary heart disease (CHD) has been described in many epidemiologic and well designed prospective trials. Since first being elucidated by the Coronary Primary Prevention Trial, reducing levels of blood cholesterol results in a corresponding reduction in CHD risk has been demonstrated by numerous trials. The evidence now indicates that cholesterol reduction by any number of means confers up to a 35% reduction in total mortality, coronary mortality, coronary artery procedures, stroke, and other CHD-related events. This article reviews data that demonstrate cholesterol reduction decreases CHD risk, discusses current and emerging treatment modalities, and describes the methods healthcare practitioners can use to enhance lipid treatment outcomes. It also identifies educational tools that can be used to empower patients to improve their compliance and become actively involved in reducing their CHD risk. PMID- 12572626 TI - A novel therapeutic approach to dyslipidemia. AB - The National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) guidelines call for more aggressive lowering of low-density lipoprotein cholesterol (LDL-C) and will substantially increase the number of patients eligible for lipid-lowering therapy. Statins, the current treatment standard, have proven to be highly efficacious in lowering LDL-C and reducing coronary heart disease (CHD) risk. Because some patients are unable to tolerate 3-hydroxy 3-methylglutaryl coenzyme A reductase inhibitors (statins) or they are not candidates for statin therapy, however, other cholesterol-lowering modes of therapy are needed. Currently available nonstatin drugs often do not reliably reduce LDL-C to a desired extent or are limited in their safety and tolerability. Ezetimibe, a novel lipid-lowering agent until recently in phase III development, is the first in a new class of selective cholesterol absorption inhibitors and offers a promising new approach to the treatment of dyslipidemia. This article reviews the cholesterol metabolic pathways and the mechanism of action of the currently available lipid-modifying agents and introduces ezetimibe, the first selective cholesterol absorption inhibitor. PMID- 12572627 TI - Palm technology moves resident data recording into new dimension. PMID- 12572628 TI - Acknowledging coauthors' efforts. PMID- 12572629 TI - Training needed to dispel negative stereotype of elderly patients. PMID- 12572630 TI - DOs should embrace increased emphasis on care for older adults. PMID- 12572631 TI - Results of a survey of inaugural class graduates of a college of osteopathic medicine. AB - The purpose of this study was to determine where the graduates of an inaugural class of a college of osteopathic medicine came from, what influenced their school selection, how their osteopathic medical school experience affected them, and how they chose what and where they would study after graduation as well as where they would practice. These data have significant implications for the osteopathic profession and its future recruitment efforts into the profession and into its postgraduate programs. PMID- 12572632 TI - Use of osteopathic manipulative treatment by Ohio osteopathic physicians in various specialties. AB - The authors mailed a survey designed to determine the use of osteopathic manipulative treatment (OMT) to the 2,318 active osteopathic physicians registered with the Ohio Osteopathic Association; 871 responses were received, for a response rate of 38%. Approximately 75% of the respondents had not or had rarely used OMT: 44% of these respondents did not use any OMT and 31% reported treating fewer than 10 patients with OMT during the week before the survey. Approximately 25% of the surveyed osteopathic physicians treated more than 10 patients with OMT, and about 6% of these treated more than 30 patients with OMT. Respondents represented 40 specialty disciplines. All of the osteopathic physicians in 17 specialties reported no OMT use, osteopathic physicians in 9 specialties reported using OMT for fewer than 10 patients during the previous week, and osteopathic physicians in 9 specialties reported using OMT for more than 10 patients during the previous week. Of the somatic dysfunctions listed in the survey, low back disorders were treated with OMT most often. Few osteopathic specialists used OMT for patients with asthma or chronic obstructive pulmonary disease. The data suggest that a great opportunity exists to increase the use of OMT by osteopathic physicians, especially those who are specialists. PMID- 12572633 TI - Where have all the heroes gone? PMID- 12572634 TI - Erectile dysfunction: prevalence, etiology, and major risk factors. AB - Erectile dysfunction is a multifactorial condition that may include psychological, neurologic, hormonal, vascular, or cavemosal impairment, or a combination of these factors. Major risk factors include aging, depression, and lifestyle. The diagnosis of erectile dysfunction can be an indicator for undiagnosed diseases, including coronary artery disease, hypertension, and diabetes mellitus. Erectile dysfunction is highly prevalent in the United States, affecting approximately 30 million men. Erectile dysfunction in the majority of men remains undiagnosed, and many men who receive treatment discontinue it because of general dissatisfaction with real-life issues that can have an impact on therapeutic success. Early identification, behavior modification, and increased therapeutic options may improve patient outcomes. PMID- 12572635 TI - Counseling the patient with erectile dysfunction: a primary care physician perspective. AB - Primary care physicians might be tempted to avoid addressing the topic of erectile dysfunction because they mistakenly believe that hectic work schedules, unforeseen reimbursement issues, or complexity of disease management preclude effectively dealing with this important disorder. Yet, it is eminently possible to complete a targeted history and physical examination within the constraints of the typical ambulatory visit. The pertinent typical medical, sexual, and psychosocial history should take about 4 to 6 minutes, and a physical examination can be done in approximately 2 to 4 minutes. Whenever feasible, partner input can be a highly valuable asset for correctly diagnosing erectile dysfunction and treating the patient. Inclusion of the partner can provide additional information, identify obstacles to success, and ensure partner readiness for resumption of coital activity. In addition, evaluation tools such as the Sexual Health Inventory for Men can be highly effective in initiating discussion on an uncomfortable subject. PMID- 12572636 TI - Therapeutic strategies for managing erectile dysfunction: a step-care approach. AB - During the past two decades, advances in the understanding of erectile dysfunction have provided primary care physicians, urologists, and other healthcare providers with additional treatment options. The introduction of sildenafil citrate, the first oral phosphodiesterase type 5 (PDE5) isoenzyme inhibitor, has helped to restore erectile function and improve overall health and quality of life. The step-care approach to the treatment of erectile dysfunction can be divided into first-, second-, and third-line modes of therapy. Most patients with erectile dysfunction will choose oral agents because of their simplicity and ease of administration. This article reviews all treatment strategies for erectile dysfunction, with a focus on PDE5 inhibitors, including novel agents that may optimize patient outcomes. PMID- 12572637 TI - Anticaries efficacy of a sodium monofluorophosphate dentifrice containing xylitol in a dicalcium phosphate dihydrate base. A 30-month caries clinical study in Costa Rica. AB - PURPOSE: To compare the long-term caries increment associated with the use of two dentifrices: (1) a test dentifrice containing 0.836% sodium monofluorophosphate (1100 ppm F) in a dicalcium phosphate dihydrate base plus 10% xylitol; and (2) a positive control dentifrice containing 0.836% sodium monofluorophosphate (1100 ppm F) in a dicalcium phosphate dihydrate base. MATERIALS AND METHODS: Three thousand three hundred ninety four (3,394) school children aged 7-12 years were selected to participate in this double blind clinical caries study at 28 public schools in the central plateau of Costa Rica. Children accepted for participation were stratified into two balanced groups within the participating schools on the basis of age and sex, and assigned to one of the two dentifrices. The study was conducted in harmony with the guidelines for caries clinical studies published in 1988 by the Council on Dental Therapeutics of the American Dental Association, and employed clinical diagnostic criteria provided in 1991 guidelines published by the National Institute of Dental Research (NIH/NIDR). Dental radiographs were not employed. RESULTS: Of those subjects who met the initial inclusion/exclusion criteria for this study, 2,539 were available for the 30-month exam. DFS increments over the 30-month period of this clinical study were 1.30 for the test dentifrice containing 10% xylitol, and 1.51 for the positive control dentifrice without xylitol; for DFT, the corresponding increments were 0.69 for the test dentifrice containing 10% xylitol, and 0.81 for that which did not contain xylitol. For both DFS and DFT, the increments associated with the test dentifrice containing 10% xylitol were statistically significantly lower than those associated with the positive dentifrice without xylitol (P < 0.05), with the observed reductions in caries increment exceeding 10% in for both parameters. PMID- 12572638 TI - Calcium phosphate precipitation method for the treatment of dentin hypersensitivity. AB - PURPOSE: To evaluate the feasibility of the calcium phosphate precipitation (CPP) method as a treatment for dentin hypersensitivity using vital teeth of beagle dogs. Also, gingival tissue response to two types of CPP methods was examined histologically. MATERIALS AND METHODS: Dentin tubules of the dogs' vital teeth were exposed by shallow cavity preparation followed by etching with 50% citric acid to simulate the condition of hypersensitive dentin. After CPP treatment was applied to the vital tooth, the dentin surface and longitudinal sections were observed by SEM to evaluate the occluding ability of the CPP method. The precipitate was also analyzed by energy-dispersive X-ray microanalysis to obtain compositional information. The gingival tissue before and after CPP method was examined histologically with light microscopy. RESULTS: Dentin tubules were occluded homogeneously and completely with an apatitic mineral after application of the CPP treatment in vital teeth. However, the depth of the precipitate in dentin tubules from the dentin surface was approximately half that seen in extracted teeth. No histological change was observed in gingival tissues when NaHCO3 was used as a post-treatment solution of the CPP method, whereas another CPP method using NaOH solution resulted in atrophy and degeneration of the epithelium of gingival tissue. PMID- 12572639 TI - Influence of microbrush on efficacy of bonding into root canals. AB - PURPOSE: Recently, several 'one-bottle' systems have been introduced for bonding into root canals. This study evaluated the influence of two brushes used as carriers of a primer-adhesive solution in the formation of resin tags, adhesive lateral branches and hybrid layer when used to bond fiber posts. MATERIALS AND METHODS: Twenty root canal treated teeth, extracted for periodontal reasons, were selected for this study. The samples were randomly divided into two groups of 10 samples each: Group 1: Microbrush + One Step + Dual Link resin cement; Group 2: One Step + Dual Link resin cement. The adhesive system and resin cement were used strictly following manufacturers' instructions. The priming-adhesive solution of the 'one-bottle' system was light-cured before placing the resin cement and the post. Twenty translucent fiber posts were used. Then, root samples were processed for SEM observation. RESULTS: The adhesive system showed resin dentin interdiffusion zone (RDIZ), resin tag and adhesive lateral branch formation. Microscopic examination of restored interfaces of Group 1 showed a higher % (P < 0.05) of RDIZ than those found in samples of Group 2. In Group 2 samples RDIZ morphology was well detectable and uniform in the first two thirds of root canals while in the apical third the RDIZ was not uniformly present while in Group 1 RDIZ was visible at the apical third too. No statistically significant differences were found among the two groups coronally and at the middle third, while the apical third of Group 1 showed significantly more resin tags than in Group 2. The characteristic reverse cone shape of resin tags was always noted in the coronal and middle third of the root canals of both groups and at the apical third of Group 1. In the apical third of Group 2 root canals, the resin tags showed a less uniform morphology and a shorter length than those found in the other observed thirds. PMID- 12572640 TI - Proximal contact formation with different restorative materials and techniques. AB - PURPOSE: To determine, whether different tooth colored materials and application techniques influence the proximal contact strengths (PCS) in vitro. MATERIALS AND METHODS: Standardized MO-cavities for direct restorations were prepared into 360 artificial lower first molars. Eight groups were formed (n = 45) and restored with amalgam (1), low (2, 3), medium (4, 5) and high (6, 7) viscosity hybrid composites and compared to the original contact strengths of the unprepared teeth (8). The restored teeth were mounted into a socket simulating physiological tooth mobility. PCS were quantified by measuring the forces necessary for passing the proximal contact with floss under standardized conditions using a universal testing machine. RESULTS: PCS ranged from 1.32 +/- 0.56 N for the low viscosity composite to 9.90 +/- 1.98 N for the Amalgam restoration. Medium (5.65 +/- 1.08 N) and high viscosity hybrid composites (5.90 +/- 1.45 N) inserted in a multi layer technique showed no statistically significant differences to the original PCS of the unprepared teeth (5.76 +/- 0.53 N). PMID- 12572641 TI - Hierarchy of pulp capping and repair activities responsible for dentin bridge formation. AB - PURPOSE: The importance of pulp capping variables which mediate pulp repair activities and dentin bridge formation following pulp exposure are not well understood. Consequently, the aim of this study was to investigate the effects of pulp capping variables on tertiary dentin bridge formation. MATERIALS AND METHODS: Calcium hydroxide [Ca(OH)2], resin-modified glass-ionomer (RMGI) and resin-based composite (RBC) were used to restore pulp exposures prepared in 161 nonhuman primate teeth, according to ISO usage guidelines. Teeth were collected from between 7 and 720 days to observe the pulp repair activities leading to dentin bridge formation. Bacteria were detected using McKays stain. Pulp activity was categorized histomorphometrically using ISO standards, and analyzed statistically using ANOVA statistics. RESULTS: The pulp capping variables in order of their relationship to dentin bridge area, from the most important to the least important were: time elapsed since pulp capping (P = 0.0009), pulp capping materials (P = 0.0252), bacterial microleakage (P = 0.0358) and area of operative debris in the dentin bridge (0.0362). A further 10 pulp capping variables were found to be less important. Relationships between pulp capping materials, operative debris, tunnel defects, bacterial microleakage, inflammation and dentin bridge formation were observed. Bacteria contaminated 18.6% of RBC, 22.2% of RMGI and 47.0% of Ca(OH)2 capped pulps. PMID- 12572642 TI - Microleakage of Class V composites using different placement and curing techniques: an in vitro study. AB - PURPOSE: To evaluate microleakage at enamel and dentin margins of two composite resins, using bulk and incremental placement techniques, "rebonding", and facial and lingual curing methods. MATERIALS AND METHODS: One hundred standardized Class V cavity preparations were made on the facial surface of extracted human premolars and were randomly assigned to 10 groups. Single Bond was used as the dentin/enamel adhesive. A heavily filled composite resin, Z250, and a microfill, Silux Plus, were inserted and polymerized using five different techniques: (1) incremental placement and facial curing; (2) incremental placement, facial curing and rebonding; (3) bulk placement and facial curing; (4) bulk placement, facial curing and rebonding; (5) incremental placement and lingual and facial curing. After the restorations were finished and polished, the margins of those in the rebonded groups were etched, rinsed, and dried. The adhesive resin, Single Bond, was applied at the composite resin-tooth interface and light-cured. All the specimens were thermocycled, stained with 1% methylene blue, sectioned, and evaluated for leakage (0-4 scale) by two examiners. RESULTS: Almost no leakage occurred at enamel margins. At the cementum margins, differences in microleakage related to restorative material or technique were not statistically significant. However, leakage at the cementum margins was significantly greater than at the enamel margins for both composite resin materials. PMID- 12572643 TI - Distribution of cariogenic bacteria in carious lesions around tooth-colored restorations. AB - PURPOSE: To compare the distribution of the three most suspected cariogenic groups of bacteria, mutans streptococci, Actinomyces naeslundii genospecies 2, and lactobacilli in carious lesions around tooth-colored restorations. MATERIALS AND METHODS: Samples from extracted teeth with clinical secondary caries around tooth-colored restorations were immuno-labeled with antibodies to the referred groups of bacteria and subsequently analyzed three dimensionally using confocal microscopy. RESULTS: The three different bacterial groups were usually present in secondary caries around tooth-colored restorations suggesting that they are involved in its formation and development. PMID- 12572644 TI - Influence of NaOCl deproteinization on shear bond strength in function of dentin depth. AB - PURPOSE: To determine the influence of NaOCl application on shear bond strengths (SBS) to superficial and deep dentin, using Prime & Bond 2.1. MATERIALS AND METHODS: Superficial and deep dentin was exposed in 40 extracted third molars by sectioning the occlusal surface immediately under the enamel-dentin junction or close to the pulp chamber. After polishing the dentin disks (600-grit SiC), they were assigned to two groups: (1) 36% orthophosphoric acid for 15 seconds, or (2): 36% orthophosphoric acid for 15 seconds followed by 5% sodium hypochlorite (NaOCl) for 2 minutes. The dentin adhesive was applied as per manufacturer's instructions, followed by TPH resin-based composite. The specimens were stored in water for 24 hours at 37 degrees C and thermocycled x500 and SBS were determined. Data were statistically analyzed with ANOVA and Student's t-test. RESULTS: For acid-etched specimens, superficial dentin resulted in statistically higher mean SBS than deep dentin. After NaOCl application, deep and superficial dentin resulted in statistically similar mean SBS. Collagen removal did not affect mean SBS on superficial dentin, while SBS values on deep dentin were increased. PMID- 12572645 TI - The effects of outward forced convective flow on inward diffusion of potassium across human dentin. AB - PURPOSE: To measure the inward flux of potassium across human dentin disks, in vitro, in the absence and then in the presence of a simulated pulpal pressure (PP), before and after brushing with two desensitizing dentifrices. MATERIALS AND METHODS: Dentin discs were made from extracted unerupted third molars and etched with 6% citric acid to remove the smear layers. The hydraulic conductance (Lp or outward fluid movement) of the discs was measured before and after brushing for 2 minutes with two desensitizing dentifrices, Colgate Sensitive Maximum Strength or Sensodyne Fresh Mint, both dentifrices containing 5% potassium nitrate. The potassium flux was measured at a pressure of 0 cm H2O, and a simulated pulpal pressure of 20 cm H2O. RESULTS: The results showed the Colgate-brushed specimens had lower Lp (P < 0.05) than specimens brushed with Sensodyne or with saline. The Colgate-brushed specimens had a corresponding lower K+ flux, at a PP = 0 cm H2O, than specimens brushed with saline, but were not significantly different than specimens brushed with Sensodyne dentifrice. However, when the pressure was increased from 0 to 20 cm H2O, to simulate outward dentin fluid pressure, there were significant decreases in K+ flux across all three treatment groups, but the K+ flux of the Colgate-brushed specimens were affected the least, resulting in the Colgate-brushed specimens having significantly (P < 0.05) greater K+ fluxes than the saline or Sensodyne dentifrice-brushed groups. The magnitude of reduction in K+ flux in going from 0 cm H2O to 20 cm H2O was demonstrated to be linearly related to the magnitude of the Lp of the brushed specimens. Hence, a small Lp corresponded to a small reduction in K+ flux, and a correspondingly large K+ flux at 20 cm H2O. An explanation of this phenomenon is that a decrease in the Lp of dentin, produced by brushing, corresponds to a decreased outward dentin fluid flow. This lowers the outward K+ convective flow and also lowers the inward K+ diffusion. However, since the decrease of the outward K+ convective flow is greater than the decrease of the inward K+ diffusion, and the net flux is the algebraic sum of these two terms, the result is greater net inward K+ flux through the dentin. Therefore, the Colgate-brushed specimens, which exhibited the lowest Lp, provided the greatest inward K+ flux at simulated dentin fluid pressure. PMID- 12572646 TI - Microleakage of amalgam cavity treatment systems: an in vitro evaluation. AB - PURPOSE: to evaluate the effectiveness of four cavity treatment systems for their ability to seal standardized Class V restorations of amalgam. MATERIALS AND METHODS: The Celay copy-milling machine was used to prepare twenty-five standardized Class V cavities on the buccal and palatal surfaces of maxillary premolar teeth extracted for orthodontic purposes. The cavosurface margins lay entirely within enamel. Five cavity treatment test groups were employed prior to amalgam placement: Group 1, Cervitec; Group 2, Gluma One Bond; Group 3, Panavia 21; Group 4, Copaliner Dentin Varnish and Sealant and Group 5, Control. Following restoration placement, the teeth were aged at room temperature (25 +/- 2 degrees C) in tap water for 2 months. The specimens were subsequently thermocycled (500 cycles, 5 -55 +/- 2 degrees C), immersed in a dye solution, sectioned and scored for microleakage. Scanning electron microscopy was employed to examine features of the tooth/restoration interface. RESULTS: None of the materials tested consistently prevented microleakage. No statistically significant differences were identified between the groups with respect to the microleakage scores (P > 0.05). The gingival walls exhibited significantly more microleakage than the occlusal walls (P = 0.00). No significant differences were identified between the microleakage scores obtained for the buccal and palatal cavities. Scanning electron microscopy examination failed to demonstrate a consistent pattern of interfacial appearance, showing a combination of cohesive and adhesive failure within and between test groups. PMID- 12572647 TI - Laser fluorescence detection of occlusal caries. AB - PURPOSE: This study compared six diagnostic methods with a histologic standard, including diode laser fluorescence, caries detection dye, and an operating microscope. MATERIALS AND METHODS: Diagnosis groups included: visual only (VO); visual and bitewing radiograph (VX); visual, bitewing radiograph and explorer (VXE); Global Protege operating microscope at x10 (S); caries detection dye (CDD); diode laser fluorescence (DLF). Three examiners individually evaluated five independent sites on 29 extracted human molars using the above methods. Teeth were sectioned and evaluated histologically. Response Operating Characteristic (ROC) analysis was used to descriptively and inferentially assess the relationship between occlusal caries and the six diagnostic methods. RESULTS: In this in vitro study, the resulting AUCs (areas under the ROC curve) showed that the DLF method appeared to be superior, when compared to traditional methods (VO, VX, VXE) and caries detection dye. While caries detection with DLF ranked higher than use of an operating scope, they were not statistically different although the difference tended toward significance (P = 0.0511). Furthermore, the AUC for CDD would be ranked lower in accuracy than the other diagnostic methods tested and could result in over treatment of sound teeth. DLF use offers improved detection of suspected hidden caries that have not become radiographically evident. PMID- 12572648 TI - A 3-year clinical evaluation of a compomer, a composite and a compomer/composite (sandwich) in class II restorations. AB - PURPOSE: To compare the clinical performance of Dyract, Spectrum-TPH and Dyract layered with Spectrum-TPH (sandwich) in Class II restorations of permanent dentitions. MATERIALS AND METHODS: 23 adult patients aged 25-62 were selected at random in a general dental practice. Each patient had 3 bicuspid teeth restored with Dyract, Spectrum-TPH and a "sandwiched" combination where Dyract was covered by the composite resin TPH. During the entire study, three patients were lost. Restorations were clinically evaluated using a modified Ryge/USPHS system. RESULTS: Over 36 months all three types of restorations performed well clinically. No post-operative sensitivity was reported by any patient indicating proper bonding and sealing. Only one Dyract/TPH sandwich restoration had to be replaced after 2.5 years due to root caries. The Dyract restorations exhibited significantly higher (P = 0.0039) occlusal wear than the TPH and the Dyract/TPH sandwiched restorations. The marginal integrity was also found to be significantly better (P = 0.001) for the TPH and Dyract/TPH restorations compared to Dyract alone, while no significant difference (P > 0.05) could be demonstrated between TPH and Dyract/TPH restorations. The low rate of failure in the three different restorative systems suggests that they are reliable restorative materials in permanent bicuspids over a 3-year period. PMID- 12572649 TI - Can modern restorative procedures and materials reliably seal cavities? In vitro investigations. Part 2. AB - Secondary caries, marginal discoloration and marginal gap/fracture account for a significant majority of clinical diagnoses of restoration failure and reason for restoration replacement. These failures occur at the interface between the restoration and the cavity preparation. It is an underlying belief, or at least an underlying assumption of the dental profession, that improving the seal between a restorative material and tooth structure will reduce the frequency of such occurrences, which will in turn lead to enhanced restoration longevity. This paper reviews the current status of in vitro literature regarding the ability of modern restorative materials and techniques to reliably seal cavity preparations. In addition, methodology for accomplishing in vitro research is reviewed for the purpose of elucidating trends, areas of interest and controversial topics. PMID- 12572650 TI - Effectiveness of Sonicare power toothbrush to remove chlorhexidine stains. AB - PURPOSE: To describe the effect of Sonicare power toothbrush on removing chlorhexidine stains from human teeth. MATERIALS AND METHODS: Two cases with chlorhexidine staining were treated with Sonicare power toothbrush for a 4-week period. Clinical evaluations occurred at baseline, 2 weeks from baseline and 4 weeks from baseline. Stain level was assessed with the Lobene Tooth Stain Index. The labial surface of each stained incisor or canine was divided into two regions: margin and body. RESULTS: Sonicare reduced the average area of stain score by 58.5-63.3% after 2 weeks and 68.3-76.7% after 4 weeks. The average intensity of stain scores was reduced by 50.0-56.5% and 58.9-69.6% after 2 and 4 weeks, respectively. PMID- 12572651 TI - Principles of cell mechanics for cartilage tissue engineering. AB - The critical importance of mechanical signals to the health and maintenance of articular cartilage has been well demonstrated. Tissue engineers have taken a cue from normal cartilage physiology and incorporated the use of mechanical stimulation into their attempts to engineer functional cartilage. However, the specific types of mechanical stimulation that are most beneficial, and the mechanisms that allow a chondrocyte to perceive and respond to those forces, have yet to be elucidated. To develop a better understanding of these processes, it is necessary to examine the mechanical behavior of the single chondrocyte. This paper reviews salient topics related to chondrocyte biomechanics and mechanotransduction, and attempts to put this information into a context both appropriate and useful to cartilage tissue engineering. It also describes the directions this exciting field is taking, and lays out a vision for future studies that could have a significant impact on our understanding of cartilage health and disease. PMID- 12572652 TI - Adaptations of trabecular bone to low magnitude vibrations result in more uniform stress and strain under load. AB - Extremely low magnitude mechanical stimuli (<10 microstrain) induced at high frequencies are anabolic to trabecular bone. Here, we used finite element (FE) modeling to investigate the mechanical implications of a one year mechanical intervention. Adult female sheep stood with their hindlimbs either on a vibrating plate (30 Hz, 0.3 g) for 20 min/d, 5 d/wk or on an inactive plate. Microcomputed tomography data of 1 cm bone cubes extracted from the medial femoral condyles were transformed into FE meshes. Simulated compressive loads applied to the trabecular meshes in the three orthogonal directions indicated that the low level mechanical intervention significantly increased the apparent trabecular tissue stiffness of the femoral condyle in the longitudinal (+17%, p<0.02), anterior posterior (+29%, p<0.01), and medial-lateral (+37%, p<0.01) direction, thus reducing apparent strain magnitudes for a given applied load. For a given apparent input strain (or stress), the resultant stresses and strains within trabeculae were more uniformly distributed in the off-axis loading directions in cubes of mechanically loaded sheep. These data suggest that trabecular bone responds to low level mechanical loads with intricate adaptations beyond a simple reduction in apparent strain magnitude, producing a structure that is stiffer and less prone to fracture for a given load. PMID- 12572653 TI - Volume currents in forward and inverse magnetoencephalographic simulations using realistic head models. AB - Volume currents are important for the accurate calculation of magnetoencephalographic (MEG) forward or inverse simulations in realistic head models. We verify the accuracy of our finite element method implementation for MEG simulations by comparing its results for spheres containing dipoles to those obtained from the analytic solution. We then use this finite element method to show that, in an inhomogeneous, nonspherical realistic head model, the magnetic field normal to the MEG detector due to volume currents often has a magnitude on the same order or greater than the magnitude of the normal component of the primary magnetic field from the dipole. We also demonstrate the disparity in forward solutions between a model that employs spheres, one that uses the realistic head and primary currents alone, and a realistic head model that incorporates both primary and volume currents. In forward and inverse MEG simulations using the inhomogeneous realistic model, the results obtained from calculations containing volume currents are more accurate than those derived without considering volume currents. PMID- 12572654 TI - Simultaneous electrical and optical mapping in rabbit hearts. AB - Local cardiac excitation is measured from either extracellular voltage (Ve) or optical transmembrane voltage (Vm) with fluorescent dye. We used a transparent electrode array and a laser scanner to test whether Ve and coepicentral optical Vm give equivalent excitation times during epicardial pacing and sinus beats. To help explain time differences, we estimated interrogation width/depth ratios for Ve and optical Vm with dipole and Monte Carlo models. Magnitudes of time differences between excitation measured with Ve and optical Vm during pacing were 1.9 +/- 1.9 ms (n= 1,112 recording pairs). Deeper interrogation for optical Vm vs Ve was indicated with sinus beats that contained a transmural propagation component. When pacing produced propagation along epicardial fibers, excitation time measured from optical Vm was later than excitation time measured with epicardial Ve. When pacing produced propagation across epicardial fibers, excitation time measured from optical Vm was earlier than that measured with epicardial Ve. These time differences could be attributed to deeper optical interrogation and fiber rotation with depth in ventricles. PMID- 12572655 TI - Progress towards patient-specific computational flow modeling of the left heart via combination of magnetic resonance imaging with computational fluid dynamics. AB - A combined computational fluid dynamics (CFD) and magnetic resonance imaging (MRI) methodology has been developed to simulate blood flow in a subject-specific left heart. The research continues from earlier experience in modeling the human left ventricle using time-varying anatomical MR scans. Breathing artifacts are reduced by means of a MR navigator echo sequence with feedback to the subject, allowing a near constant breath-hold diaphragm position. An improved interactive segmentation technique for the long- and short-axis anatomical slices is used. The computational domain is extended to include the proximal left atrium and ascending aorta as well as the left ventricle, and the mitral and aortic valve orifices are approximately represented. The CFD results show remarkable correspondence with the MR velocity data acquired for comparison purposes, as well as with previously published in vivo experiments (velocity and pressure). Coherent vortex formation is observed below the mitral valve, with a larger anterior vortex dominating the late-diastolic phases. Some quantitative discrepancies exist between the CFD and MRI flow velocities, owing to the limitations of the MR dataset in the valve region, heart rate differences in the anatomical and velocity acquisitions, and to certain phenomena that were not simulated. The CFD results compare well with measured ranges in literature. PMID- 12572656 TI - Relation between near-wall residence times of monocytes and early lesion growth in the rabbit aorto-celiac junction. AB - Transient particle-hemodynamic simulations were conducted in a model of the rabbit aorto-celiac junction to investigate mechanisms responsible for localized monocyte attachment and subsequent lesion formation. We hypothesized that the probability for monocyte deposition is related to discrete near-wall particle stasis and/or elevated concentrations, as encapsulated by a new near-wall residence time (NWRT) parameter. A low wall shear stress (WSS) condition accounted for factors such as endothelial cell expression of adhesive molecules as well as a reduced probability of monocyte rolling and detachment. To accurately simulate particle transport, terms for the near-wall drag modification and lift were included. Low WSS and high oscillatory shear index parameters proved ineffective compared to localized in vivo results of monocyte accumulation and lesion initialization. The NWRT parameter, with a limiting WSS condition, identified the lateral flow divider as most susceptible to monocyte deposition, as observed in vivo. A representative quantitative correlation between monocyte deposition and NWRT occurrence was established (r2 = 0.77 and p<10(-4)) on a highly focal basis for an averaged data set. Results indicate that cell transport and conditions for hemodyamically induced surface reactivity are necessary components in formulating an effective model for monocyte adhesion in complex three-dimensional vessel configurations. PMID- 12572657 TI - Nitric oxide delivery system for cell culture studies. AB - To investigate the toxicity and mutagenicity of NO, methods are needed to deliver it to cell cultures at known, constant rates. To permit continuous exposures over lengthy periods, we fabricated a simple apparatus utilizing gas-permeable polydimethylsiloxane (Silastic) tubing to supply both NO and O2 to a stirred, cylindrical vessel. Mass transfer in this system was characterized by measuring the delivery rates of NO or O2 alone, and of NO to air-saturated solutions. The concentrations of NO, O2, and NO2- (the end product of NO oxidation) were monitored continuously. The total flux of nitrogen species into the liquid (as determined from the sum of NO and NO2- accumulation) was 50%-90% greater in the presence of O2, depending on the NO partial pressure in the gas. Also, the simultaneously measured mass transfer coefficients for NO and O2 differed greatly from the corresponding unreactive values. An analysis of the data using diffusion reaction models showed that NO oxidation in the aqueous boundary layer contributed very little to the nitrogen flux increase or to variations in the mass transfer coefficients. However, the unusually strong dependence of the delivery rates on chemical reactions could be explained by postulating that partial oxidation of NO to NO2 occurred within the membrane. The rate constant we estimated for polydimethylsiloxane, 4.4 x 10(5) M-2 s(-1) at 23 degrees C, is only about one-fifth of values reported previously for water and nonpolar solvents, but the high solubilities of NO and O2 in the polymer are sufficient to make NO2 formation significant. Although considerable NO2 is calculated to enter the liquid, its reaction with aqueous NO is rapid enough to keep this undesired compound at trace levels, except within a few microns of the tubing. Thus, cells will have little exposure to NO2 PMID- 12572658 TI - Viability of cancer cells exposed to pulsed electric fields: the role of pulse charge. AB - The goal of this study was to collect a comprehensive set of data that related lethal effects of electric fields to the duration of the pulse. Electric pulses of different strengths and durations were applied to a suspension of HEp-2 cells (epidermoid carcinoma of the human larynx) using a six-needle electrode array connected through an autoswitcher to a square wave generator. Pulse durations varied from 50 micros to 16 ms and the ranges of electric field were adjusted for each duration to capture cell viabilities between 0% and 100%. After pulsation, cells were incubated for 44 h at 37 degrees C, and their viability was measured spectrophotometrically using an XTT assay. For each pulse duration (d), viability data were used to determine the electric field that killed half of the cells (E50). When plotted on logarithmic axes, E50 vs. d was a straight line, leading to a hyperbolic relationship: E50=const/d. This relationship suggests that the total charge delivered by the pulse is the decisive factor in killing HEp-2 cells. PMID- 12572659 TI - Frequency characterization of blood glucose dynamics. AB - Examples of the frequency range of blood glucose dynamics of normal subjects and subjects with diabetes are reported here, based on data from the literature. The frequency band edge was determined from suitable, frequently sampled blood glucose recordings using two methods: frequency domain estimation and signal reconstruction. The respective maximum acceptable sampling intervals, or Nyquist sampling periods (NSP), required to accurately represent blood glucose dynamics were calculated. Preliminary results based on the limited data available in the literature indicate that although blood glucose NSP values are higher in most diabetic subjects, values in some diabetic subjects are indistinguishable from those of normal subjects. High fidelity monitoring sufficient to follow the intrinsic blood glucose dynamics of all diabetic subjects requires a NSP of approximately 10 min, corresponding to a continuous frequency band edge of approximately 1 x 10(-3) Hz. This analysis provides key information for the design of clinical studies that include blood glucose dynamics and for the design of new glucose monitoring systems. PMID- 12572660 TI - An evaluation of population D-optimal designs via pharmacokinetic simulations. AB - One goal of large scale clinical trials is to determine how a drug is processed by, and cleared from, the human body [i.e., its pharmacokinetic (PK) properties] and how these PK properties differ between individuals in a population (i.e., its population PK properties). Due to the high cost of these studies and the limited amount of data (e.g., blood samples) available from each study subject, it would be useful to know how many measurements are needed and when those measurements should be taken to accurately quantify population PK model parameters means and variances. Previous studies have looked at optimal design strategies of population PK experiments by developing an optimal design for an individual study (i.e., no interindividual variability was considered in the design), and then applying that design to each individual in a population study (where interindividual variability is present). A more algorithmically and informationally intensive approach is to develop a population optimal design, which inherently includes the assessment of interindividual variability. We present a simulation-based evaluation of these two design methods based on nonlinear Gaussian population PK models. Specifically, we compute standard individual and population D-optimal designs and compare population PK model parameter estimates based on simulated optimal design measurements. Our results show that population and standard D-optimal designs are not significantly different when both designs have the same number of samples per individual. However, population optimal designs allow for sampling schedules where the number of samples per individual is less than the number of model parameters, the theoretical limit allowed in standard optimal design. These designs with a low number of samples per individual are shown to be nearly as robust in parameter estimation as standard D-optimal designs. In the limit of just one sample per individual, however, population D-optimal designs are shown to be inadequate. PMID- 12572661 TI - Parallels and contrasts between iron and copper metabolism. AB - This paper reviews the Second International Workshop on Iron and Copper Homeostasis, held in Pucon, Chile 10-13 November, 2001. We cover the presentations and papers published (this issue) with the intent to point out parallels, contrasts and cutting edge areas rather than to say something about every paper. Iron and copper metabolism have been intertwined for nearly 150 years and the interrelationship is growing with advances in understanding the role of ceruloplasmin as one example and the probable role of hephaestin as another. The transporter DMT1 (divalent metal transporter 1) clearly plays a major part in iron uptake and trafficking. Emerging evidence suggests that it plays a lesser role in manganese, cadmium and copper transport; but it is still being evaluated there. Yet another interaction may come from the IRE/IRP (Iron Responsive Element/Iron Regulatory Protein) story where a paradigmatic role in iron homeostasis is well established, but interaction with copper is only now emerging. Parallels include the nutrient status of both metals based on their utility for redox reactions as well as their toxicity primarily via reactive oxygen species. The workshop also revealed that alternate splicing of pre-mRNAs for iron and copper related proteins and tissue specific responses are additional similarities. Regulation of gene expression and excretion offered contrasts between the two metals. The workshop also considered a series of continuing and emerging issues. PMID- 12572662 TI - The copper-iron chronicles: the story of an intimate relationship. AB - During the last decade there has been a surge of interest and activity in exploring the metabolic links between copper and iron. This review describes more than a century and a half of effort that has led to our current understanding. Particular attention is given to the early events since these are less well-known and appreciated. The landmark 1928 paper of Hart, Elvehjem and coworkers is generally given credit for the start of the copper/iron field, and specifically for the discovery of the role of copper in forming hemoglobin and in overcoming anemia. However, some credit for the ideas, observations, and experiments should be shared with several investigators of the previous century. These scientists and physicians were primarily motivated to find the causes and cures of chlorosis, a common form of anemia at the time. From his chemical determination of copper in red blood cells in 1848, Millon proposed a form of chlorosis due to copper deficiency. Likewise, Pecholier and Saint-Pierre, observing the robust health of young women working in copper factories, concluded that copper was helpful in preventing and overcoming chlorosis. The first direct experimental evidence for the theory was provided by the Italian physician Mendini, who in 1862 reported that supplementation of the diet with copper salts overcame chlorosis in young women. In the 1890s Cervello and his students in Italy, using semi-quantitative hematological measurements, confirmed the beneficial effects of copper on anemia both in patients and in animal models. There was nearly a 30 year period of inactivity, but the decade of the 1930s saw renewed interest and exciting developments in the field. The Elvehjem report of 1928 was quickly verified and extended by multiple laboratories on four continents. In the 1950s and 1960s Wintrobe and Cartwright and their colleagues localized, and started to systematically evaluate, the potential sites at which copper was likely to effect iron for hemoglobin synthesis, namely, intestinal absorption, release from storage, and cellular utilization during synthesis. The copper/iron connection also has a 'flip-side', i.e., iron status can influence copper metabolism as first described by Warburg and Krebs in 1927. Thus, there are opportunities for feedback mechanisms at the cellular and physiological level that are not yet understood. The evaluation of these processes continues to this day, aided by modern molecular and genetic approaches. Studies of two copper proteins, ceruloplasmin and its recently discovered homologue hephaestin, have provided two molecular links connecting the pathways of copper and iron metabolism. The recent identification of other proteins of iron and copper metabolism, for example, copper ATPases and the membrane iron transporters DCT1/DMT1/Nramp2 and IREG1/MTP1/ferroportinl, are likely to fill crucial pathway gaps. The ongoing discovery of genes and gene mutations involved in the metabolism of copper and iron provides an important key to a deeper understanding of the connections between the pathways, and their physiological and pathological consequences. It is hoped that this historical review, by illuminating the complex paths that have led to the current state of knowledge, will contribute to our appreciation, our understanding, and perhaps our continued discovery of the connections between copper and iron. PMID- 12572663 TI - DMT1: a mammalian transporter for multiple metals. AB - DMT1 has four names, transports as many as eight metals, may have four or more isoforms and carries out its transport for multiple purposes. This review is a start at sorting out these multiplicities. A G185R mutation results in diminished gastrointestinal iron uptake and decreased endosomal iron exit in microcytic mice and Belgrade rats. Comparison of mutant to normal rodents is one analytical tool. Ectopic expression is another. Antibodies that distinguish the isoforms are also useful. Two mRNA isoforms differ in the 3' UTR: +IRE DMT1 has an IRE (Iron Responsive Element) but -IRE DMT1 lacks this feature. The +/-IRE proteins differ in the distal 18 or 25 amino acid residues after shared identity for the proximal 543 residues. A major function is serving as the apical iron transporter in the lumen of the gut. The +IRE isoform appears to have that role. Another role is endosomal exit of iron. Some evidence indicts the -IRE isoform for this function. In our ectopic expression assay for metal uptake, four metals--Fe2+, Mn2+, Ni2+ and Co2+--respond to the normal DMT1 cDNA but not the G185R mutant. Two metals did not--Cd2+ and Zn2+--and two--Cu2+ and Pb2+--remain to be tested. In competition experiments in the same assay, Cd2+, Cu2+ and Pb2+ inhibit Mn2+ uptake but Zn2+ did not. In rodent mutants, Fe and Mn appear more dependent on DMT1 than Cu and Zn. Experiments based on ectopic expression, specific antibodies that inhibit metal uptake and labeling data indicate that Fe3+ uptake depends on a different pathway in multiple cells. Two isoforms localize differently in a number of cell types. Unexpectedly, the -IRE isoform is in the nuclei of cells with neuronal properties. While the function of -IRE DMT1 in the nucleus is speculative, one may safely infer that this localization identifies new role(s) for this multifunctional transporter. Management of toxic challenges is another function related to metal homeostasis. Airways represent a gateway tissue for metal entry. Preliminary evidence using specific PCR primers and antibodies specific to the two isoforms indicates that -IRE mRNA and protein increase in response to exposure to metal in lungs and in a cell culture model; the +IRE form is unresponsive. Thus the -IRE form could be part of a detoxification system in which +IRE DMT1 does not participate. How does iron status affect other metals' toxicity? In the case of Mn, iron deficiency may enhance cellular responses. PMID- 12572664 TI - Pretranslational control of Menkes disease gene expression. AB - The gene for Menkes disease codes for a Cu-transporting ATPase that regulates Cu homeostasis in all tissues with the exception of adult liver. The basis for developmental or tissue-specific regulation at present is not understood. To learn if the regulation is associated with the promoter, we cloned and sequenced a 2.2 kb genomic DNA fragment flanking exon 1. When ligated into a pGL2 luciferase reporter gene construct, the 2.2 kb showed promoter activity, but not nearly to the extent of a 1.3 kb fragment previously reporter by Levinson et al. Sequence analysis of the nucleotides spanning the region between 1.3 kb and 2.2 kb revealed a 13-nucleotide motif ACACAAAAAAATA 2059 bp upstream from the start site that duplicated the 'hunchback' binding site, a key site controlling developmental gene expression in Drosophila. Eliminating 129 bp containing the hunchback site (Hb) from the 5' end of the 2.2 kb stimulated promoter activity, suggesting the Hb site was basically suppressive. When ligated upstream of an SV40 and tested in SY5Y cells, however, the SV40 promoter activity was strongly stimulated, which conflicts with the site being suppressive. Mutating the site in the 2.2 kb weakened the promoter activity in SY5Y and HepG2 cells and a fragment with mutated sequence ligated upstream of the SV40 cancelled the activation of SV40 promoter activity. All data suggested the Hb site was a positive controlling site for Cu-ATPase expression. Nuclear extracts from SY5Y and HepG2 cells were observed to bind to a 106 bp probe with the Hb site in a gel-shift assay. Only SY5Y proteins, however, showed a slower moving shift band indicative of a secondary interaction. A probe with mutated sequences displayed the same shift pattern, suggesting other sites in the 106 bp DNA strand were also recognizing the nuclear proteins. A Southwestern analysis suggested that proteins of 125 kD, 70 kD, 50 kD and 42 kD bound to the wild type probe; a 60 kD and all with the exception of the 42 kD bound to the mutant probe. The data support the conclusion that the distal promoter of the Menkes disease gene contains elements that interact in combinatorial fashion to regulate Cu-ATPase expression and that tissue specificity may lie with the quantity or types of distinct DNA binding proteins in the nucleus. PMID- 12572665 TI - Brain iron uptake and homeostatic mechanisms: an overview. AB - Timely and adequate iron acquisition by the brain is essential to normal neurological function. Despite the numerous cognitive and neurological impairments that are associated with disruptions in brain iron acquisition, including both too much and too little iron, the mechanism and regulation of the mechanisms by which the brain acquires iron are poorly understood. In this article, we review the current state of knowledge regarding expression of iron transport proteins in the brain, brain iron uptake and discuss why a model for brain iron uptake must take into consideration the potentially competing influences on the endothelial cell between the status of iron in the brain versus the systemic iron status. PMID- 12572666 TI - Adventiously-bound redox active iron and copper are at the center of oxidative damage in Alzheimer disease. AB - Central to oxidative damage in Alzheimer disease is the production of metal catalyzed hydroxyl radicals that damage every category of macromolecule. Studies on redox-competent copper and iron indicate that redox activity in Alzheimer disease resides exclusively within the cytosol of vulnerable neurons and that chelation with deferoxamine or DTPA removes this activity. We have also found that while proteins that accumulate in Alzheimer disease such as tau, amyloid beta, and apolipoprotein E possess metal-binding sites, metal-associated cellular redox activity is more dependent on metal-nucleic acid binding. Consistent with this finding is the large amount of cytoplasmic RNA in pyramidal neurons. Still, the source of metal-catalyzed redox activity is controversial. Heme oxygenase-1, an enzyme that catalyzes the conversion of heme to iron and biliverdin, is increased in Alzheimer disease suggesting increased heme turnover as a source of redox-active iron. Additionally, the role of mitochondria as a potential source of redox-active metals and oxygen radical production is assuming more prominence. In recent studies, we have found that while mitochondrial DNA and cytochrome C oxidase activity are increased in Alzheimer disease, the number of mitochondria is decreased, indicating accelerated mitochondria turnover. This finding, as well as preliminary studies demonstrating a reduction in microtubule density in neurons in Alzheimer disease suggests mitochondrial dysfunction as a potentially inseparable component of the initiation and progression of Alzheimer disease. PMID- 12572667 TI - Iron-activated iron uptake: a positive feedback loop mediated by iron regulatory protein 1. AB - The love-hate relationship between iron and living matter has generated mechanisms to maintain iron concentration in a narrow range, above and below which deleterious effects occur. At the cellular level, iron homeostasis is accomplished by the activity of the IRP proteins, which, under conditions of iron depletion, up-regulate the expression of the iron acquisition proteins TfR and DMT1. It has been shown that hydrogen peroxide activates IRP1 and that this activation mediates a potentially harmful increase in cell iron uptake. Here we show that IRP1 activity is also induced by iron-mediated oxidative stress. When cells were incubated with up to 20 microM of iron, a typical decrease in IRP1 and IRP2 activity was observed. Interestingly, when iron was further increased to 40 or 80 microM. IRP1 was reactivated in three of the four different cell lines tested, i.e., Caco-2 cells, N2A cells and HepG2 cells. In the fourth cell line (K562) IRP1 activity did not increase, but neither did it decrease. This response to iron was largely abrogated when the antioxidant N-acetyl cysteine was added along with iron to the culture medium. Thus, the effect of iron was mediated by oxidative stress. Increases in IRP1 activity were accompanied by increases in cell iron uptake, an indication that the activated IRP1 was functional in the activation of iron uptake. Hence, this iron-induced iron uptake feedback loop results in the increase of intracellular iron and increased oxidative stress. PMID- 12572668 TI - Copper reduction by copper binding proteins and its relation to neurodegenerative diseases. AB - Increasing evidence supports an important role for metals in neurobiology. In fact, copper binding proteins that form bioinorganic complexes are able to display oxidant or anti-oxidant properties, which would impact on neuronal function or in the triggering of neurodegenerative process. Two proteins related to neurodegenerative diseases have been described as copper binding proteins: the amyloid precursor protein (APP), a protein related to Alzheimer's disease, and the Prion protein (PrP), related to Creutzfeldt-Jakob disease. We used different synthetic peptides from APP and PrP sequences in order to evaluate the ability to reduce copper. We observed that APP(135-156), amyloid-beta-peptide (A beta(1 40)), and PrP(59-91) all have copper reducing ability, with the APP(135-156) peptide being more potent than the other fragments. Moreover, we identify His, Cys and Trp residues as key amino acids involved in the copper reduction of A beta, APP and PrP, respectively. We postulated, that in a cellular context, the interaction of these proteins with copper could be necessary to reduce copper on plasma membrane, possibly presenting Cu(I) to the copper transporter, driving the delivery of this metal to antioxidant enzymes. Moreover, protein-metal complexes could be the catalytic centers for the formation of reactive oxygen species involved in the oxidative damage present both in Alzheimer's and Prion disease. PMID- 12572669 TI - Prevention of free-radical mediated tissue damage and carcinogenesis induced by low-molecular-weight iron. AB - The unique model of iron-induced oxidative nephrotoxicity and renal cancer can be used to screen possible bioprotective substances in vivo. Ferric iron complexed with nitrilotriacetic acid (Fe-NTA) in this model is thought to be a tumor initiator as well as a promoter through the production of active oxygen species/free radicals. In the present paper, I have summarized the animal studies using this model of chemoprevention, and present some of new studies. PMID- 12572670 TI - Oxidative stress-mediated hepatotoxicity of iron and copper: role of Kupffer cells. AB - Iron- or copper-mediated catalysis leads to the generation of reactive oxygen species (ROS) that can attack biomolecules directly, with the consequent enhancement in membrane lipid peroxidation, DNA damage, and protein oxidation. Reactive nitrogen species (RNS) can also be formed, leading to nitration of aromatic structures in addition to the oxidative deterioration of cellular components. Kupffer cells, the resident macrophages of the liver, play significant roles in immunomodulation, phagocytosis, and biochemical attack. Upon stimulation, liver macrophages release biologically active products related to cell injury, namely, ROS, RNS, and both immunomodulatory and fibrogenic cytokines, with production of chemokines and adhesion molecules by other cells of the liver sinusoid. Iron and copper alter the functional status of Kupffer cells by enhancing their respiratory burst activity without modifying particle phagocytosis. This effect is probably due to extra O2 equivalents used in the oxidation of biomolecules and/or in the activating action of iron/copper on nitric oxide synthase, in addition to those employed by NADPH oxidase activity. Changes in gene expression of Kupffer cells may also be accomplished by iron and copper through modulation of the activity of transcription factors such as NF kappaB, which signals the production of cytotoxic, proinflammatory, or fibrogenic mediators. Thus, iron/copper-induced hepatotoxicity is a multifactorial phenomenon underlying actions due to the generation of ROS and RNS that may alter essential biomolecules with loss of their biological functions, modulate gene expression of Kupffer cells with production of cytotoxic mediators, or both. PMID- 12572671 TI - Copper-binding proteins in human erythrocytes: searching for potential biomarkers of copper over-exposure. AB - The recognition that copper is essential but also potentially toxic to humans has prompted the search for biomarkers of copper excess. The experimental approach followed here involves the isolation and subsequent characterization of copper binding molecules (CuBP) from human erythrocytes. Incubation (0-60 min) of freshly obtained erythrocytes in the presence of increasing concentrations of copper (10-50 microM; as 64Cu-histidine) led to time- and concentration-dependent uptake of the radioisotope. A near-maximal incorporation was attained after 20 min, with 45-55% of the radioactivity being recovered in 20,000 x g hemolysate supernatants (S-20). 64CuBP from S-20 were separated by size exclusion and metal affinity chromatography. Most radioactivity loaded into a Sephadex G-75 column was recovered in association with molecules of MMr greater than 60 KDa (largely accounted for by hemoglobin; Hb). Only negligible amounts of radioactive Cu were associated with metallothionein. With further purification, the higher MMr 64Cu binding fractions were resolved by Sephadex G-200 into two major peaks. The cpm/microg protein ratios of the first peak (high MMr) were proportional to the concentrations of copper presented to the erythrocytes. The second one contained mostly Hb molecules. Proteins from the first peak were concentrated in an affinity chromatography mini-column, suited to trap CuBP. The higher-affinity CuBP were eluted as a single peak which comprised around 60% of the load. An SDS PAGE analysis of such peak reveals the presence of three bands, of which two are non-hemoglobin Cu-binding proteins. The latter, whose identity remains to be established, had MMr of approximately 30 and 40 KDa, respectively. Preliminary data indicate that the two bands bind 64Cu within a range of concentrations, relevant to those expected to occur during copper over-exposure conditions. PMID- 12572672 TI - Role of nitric oxide in cellular iron metabolism. AB - Iron regulatory proteins (IRP1 and IRP2) control the synthesis of transferrin receptors (TfR) and ferritin by binding to iron-responsive elements (IREs) which are located in the 3' untranslated region (UTR) and the 5' UTR of their respective mRNAs. Cellular iron levels affect binding of IRPs to IREs and consequently expression of TfR and ferritin. Moreover, NO*, a redox species of nitric oxide that interacts primarily with iron, can activate IRP1 RNA-binding activity resulting in an increase in TfR mRNA levels. We have shown that treatment of RAW 264.7 cells (a murine macrophage cell line) with NO+ (nitrosonium ion, which causes S-nitrosylation of thiol groups) resulted in a rapid decrease in RNA-binding of IRP2, followed by IRP2 degradation, and these changes were associated with a decrease in TfR mRNA levels. Moreover, we demonstrated that stimulation of RAW 264.7 cells with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) increased IRP1 binding activity, whereas RNA binding of IRP2 decreased and was followed by a degradation of this protein. Furthermore, the decrease of IRP2 binding/protein levels was associated with a decrease in TfR mRNA levels in LPS/IFN-gamma-treated cells, and these changes were prevented by inhibitors of inducible nitric oxide synthase. These results suggest that NO+-mediated degradation of IRP2 plays a major role in iron metabolism during inflammation. PMID- 12572673 TI - The Enterococcus hirae paradigm of copper homeostasis: copper chaperone turnover, interactions, and transactions. AB - The cop operon is a key element of copper homeostasis in Enterococcus hirae. It encodes two copper ATPases, CopA and CopB, the CopY repressor, and the CopZ metallochaperone. The cop operon is induced by copper, which allows uncompromised growth in up to 5 mM ambient copper. Copper uptake appears to be accomplished by the CopA ATPase, a member of the heavy metal CPx-type ATPases and closely related to the human Menkes and Wilson ATPases. The related CopB ATPase extrudes copper when it reaches toxic levels. Intracellular copper routing is accomplished by the CopZ copper chaperone. Using surface plasmon resonance analysis, it was demonstrated that CopZ interacts with the CopA ATPase where it probably becomes copper loaded. CopZ in turn can donate copper to the copper responsive repressor CopY, thereby releasing it from DNA. In high copper, CopZ is proteolyzed. Cell extracts were found to contain a copper activated proteolytic activity that degrades CopZ in vitro. This post-translational control of CopZ expression presumably serves to avoid the accumulation of detrimental Cu-CopZ levels. PMID- 12572674 TI - Iron and copper homeostasis and intestinal absorption using the Caco2 cell model. AB - Whole body homeostasis can be viewed as the balance between absorption and excretion, which can be regulated independently. Present evidence suggests that for iron, intestinal absorption is the main site for homeostatic regulation, while for copper it is biliary excretion. There are connections between iron and copper in intestinal absorption and transport. The blue copper plasma protein, ceruloplasmin, and its intracellular homologue, hephaestin, play a role in cellular iron release. The studies reviewed here compare effects of Fe(II) and Cu(II) on their uptake and overall transport by monolayers of polarized Caco2 cells, which model intestinal mucosa. In the physiological range of concentrations, depletion of cellular iron or copper (by half) increased uptake of both metal ions. Depletion of iron or copper also enhanced overall transport of iron from the apical to the basal chamber. Copper depletion enhanced overall copper transport, but iron depletion did not. Pretreatment with excess copper also stimulated copper absorption. Plasma ceruloplasmin (added to the basal chamber) failed to enhance basolateral iron release, and Zn(II) failed to compete with Cu(II) for uptake. Neither copper nor iron deficiency altered expression of IREG1 or DMT1 (-IRE form) at the mRNA level. Thus, in the low-normal range of iron and copper availability, intestinal absorption of both metals appears to be positively related to the need for these elements by the whole organism. The two metal ions also influenced each other's transport; but with copper excess, other mechanisms come into play. PMID- 12572675 TI - The role of the placenta in iron transfer from mother to fetus and the relationship between iron status and fetal outcome. AB - During pregnancy, iron is transferred from the mother to the fetus across the placenta. The mechanism has been extensively studied. Altered iron metabolism changes transfer, but also has other consequences. In this review, we examine how the placenta adapts to altered iron supply, both in terms of changing cytokine expression and in relation to the proteins of iron transfer. Changing iron levels alters the levels of other metals, especially copper, and we review how this is related to changing function. There are also consequences to the placenta itself, to vascularisation and other aspects of the physiology. In turn, this has effects on the fetus and we review how growth and development are modified. Finally, we examine in more detail the efflux process, how it is regulated and, especially, the putative role of the placental Cu oxidase in the efflux process. As appropriate, we draw on data from humans, from animal models and from cell culture systems to illustrate the information. PMID- 12572676 TI - Copper exposure modifies the content and distribution of trace metals in mammalian cultured cells. AB - With this work, we have determined the cellular content of Cu, Fe and Zn in different cell lines, by using total reflection X-ray fluorescence spectrometry (TXRF). In addition, we examined whether cellular exposure to 100 micromoles l( 1) of Cu-His modifies the intracellular content and distribution of these trace metals. Our results indicate that all the cell lines displayed the same pattern of relative intracellular abundance of trace metals (Cu or = 150 mm Hg; n = 17) were acquired using 1H NMR spectroscopy. Orthogonal signal correction followed by principal components analysis were applied to these NMR data in order to facilitate interpretation, and the resulting chemometric models were validated using Soft Independent Modelling of Class Analogy. Using 1H NMR-based metabonomics, it was possible to distinguish low/ normal SBP serum samples from borderline and high SBP samples. Borderline and high SBP samples, however, were indiscriminate from each other. Our preliminary results showed that there was a relationship between serum metabolic profiles and blood pressure which, in part, was due to lipoprotein particle composition differences between the samples. Furthermore, our results indicated that serum pathology associated with blood pressure is apparent at SBP values > 130 mm Hg, which the WHO and ISH currently define as the limit between normal and high-normal. PMID- 12572800 TI - Electrophoresis of DNA in ionic liquid coated capillary. AB - An ionic liquid (IL) coated capillary was prepared and investigated for DNA separation. The electroosmotic flow of the capillary was reversed between pH 4.5 and 9. Below 900 base pairs the larger DNA fragment suffered more retardation in the IL coated capillary due to the increasing charge density of the fragment with size. In the presence of 4% hydroxyethylcellulose, the phiX174 DNA-Hae III digest fragments were baseline separated in both IL- and polyacrylamide-coated capillary except for the fragments of 271 and 281 base pairs; while the analysis time was shorter in the IL-coated capillary. Our experiments indicated that the IL-coated capillary could work stably in the run buffer for at least 96 h with no notable deterioration in performance. PMID- 12572801 TI - Determination of mutagenic 3-nitrobenzanthrone in diesel exhaust particulate matter by three-dimensional high-performance liquid chromatography. AB - Mutagenic 3-nitrobenzanthrone was determined in diesel exhaust particulate matter by three-dimensional high-performance liquid chromatography. Nitrophenylethyl, C18 and pyrenylethyl stationary phases were used as the first, second and third dimensions, respectively. Methanol was used as a mobile phase for the first and second dimensions, and dichloromethane was used for the third. Each column was coupled by a 6-port valve with a concentrator column. Effluent from the third dimensional column was detected by a photodiode array detector. The calibration graph showed good linearity in the range of 1-1000 ng ml(-1), and the detection limit (S/N = 3) was 1 ng ml(-1) 3-Nitrobenzanthrone could be detected within 45 min without the requirement of a clean-up procedure. 3-Nitrobenzanthrone in diesel exhaust particulate matter was detected in the range of 27-56 pg mg(-1) extract (n = 3). PMID- 12572802 TI - Quantitative liquid chromatography-mass spectrometry determination of catechins in human plasma by automated on-line extraction using turbulent flow chromatography. AB - A simple, fast and sensitive liquid chromatography-mass spectrometry (LC-MS) method with automated on-line extraction using turbulent flow chromatography (TFC) for the determination of five catechins in human plasma was developed. In this method, after on-line extraction by its injection onto an extractor column at turbulent flow, five catechins were backwashed onto a reversed phase column via on-line column switching and separated chromatographically at a laminar flow of 1 ml min(-1). Using this tandem LC-LC-MS system, the extraction, the separation and the quantitation of five catechins in human plasma could be achieved with satisfactory selectivity and sensitivity. The limit of detection (S/N = 3) ranged from 0.6 to 2 ng ml(-1). The described procedure was very simple and rapid since no off-line sample preparation was required, total analysis time being 18.5 min. PMID- 12572803 TI - Internal standard signal suppression by co-eluting analyte in isotope dilution LC ESI-MS. AB - The suppression of the internal standard by increasing concentrations of the co eluting analyte in calibration series and plasma samples analysed by LC-ESI-MS was studied using the isotope dilution technique. A series of three analyte/deuterated analyte pairs including fexofenadine/d6-fexofenadine, dapsone/d4-dapsone and peudoephedrine/d3-ephedrine were investigated. Suppression of the internal standard signal was noticed in extracted plasma samples containing fexofenadine and d6-fexofenadine as internal standard, as well as in solvent based calibration solutions of the three pair of compounds noted above during LC-ESI-MS analysis at flow rates greater than 100 microL min(-1). This signal suppression effect was described by invoking Enke's model of electrospray ion generation. This model suggests that signal suppression can be ascribed to the competition between ionic species for charged surface sites present on the generated droplets during the electrospray process. The slopes of the calibration curves of the three analytes were close to unity (fexofenadien/d6-fexofenadine 0.964 +/- 0.008, pseudoephedrine/d3-ephedrine 1.02 +/- 0.080 and dapsone/d4 dapsone 0.905 +/- 0.048) as predicted by the model, indicating that quantitation should not be affected by the variation in the peak area of the internal standard. PMID- 12572804 TI - A pulsed corona discharge switchable high resolution ion mobility spectrometer mass spectrometer. AB - A pulsed corona discharge ionisation source, a candidate replacement for 63Ni ionisation sources for ion mobility spectrometry, is described along with a new design of ion mobility spectrometer-mass spectrometer. Preliminary research on the characterisation of the reactant ion peaks associated with the use of this ionisation source was undertaken by assembling a pulsed corona discharge ionisation switchable high-resolution ion mobility spectrometer-mass spectrometer to enable the mobility spectra, atmospheric chemical ionisation mass spectra and selected-mass mobility spectra to be obtained. With ammonia doping at 2.39 mg m( 3) in air and a water content of approximately 80 mg m(-3) in the positive mode the observed response was attributable to the formation of 1(H2O)(n)NH4]+ and [(H2O)n(NH3)NH4]+ in the reaction region. The observed responses in the negative mode were more complex with evidence for the formation [(H2O)(n)O2]-, [(H2O)(n)CO3]-, [(H2O)(n)HCO3]-, [(H2O)(n)CO4]- and [(H2O)(n)NO3]-. The responses due to these species were clearly discernible in the resultant mobility spectra, with enough oxygen-based species formed to support analytically useful responses. PMID- 12572805 TI - Membrane introduction mass spectrometry for monitoring complexation equilibria of beta-cyclodextrin with substituted benzenes. AB - Membrane introduction mass spectrometry (MIMS) was used to monitor complexation reactions between beta-cyclodextrin (CD) and a series of benzene derivatives in aqueous solution. The equilibrium constants for benzene, chlorobenzene, bromobenzene, iodobenzene, toluene, cyanobenzene and nitrobenzene were determined. The suitability of MIMS for monitoring complexation reactions of organic compounds with host molecules was demonstrated. Structure-activity relationship analysis shows that the inclusion phenomena are driven by a variety of chemical forces, of which hydrophobicity is predominant for non-polar compounds, but not the only factor for more polar ones. PMID- 12572806 TI - Development of a microparticle-based on-site immunoassay for the detection of atrazine in soil and water samples. AB - Atrazine is widely used as a herbicide in agriculture and has been identified as a major groundwater contaminant in the US. Because of the possible hazard associated with its usage, there is a need for an efficient and economic screening method for on-site field testing of atrazine and other s-triazine herbicides in soil and water. We have developed a rapid, on-site test for the detection of atrazine based on the principle of microparticle agglutination inhibition immunoassay. The test detects 50 microg kg(-1) (0.050 ppm) atrazine in soil samples with direct extraction and 1.0 microg L(-1) atrazine in water samples when coupled with solid phase extraction. PMID- 12572807 TI - Molecular sensors using a resonance Raman template. AB - The potential use of resonance Raman spectroscopy as a molecular sensing tool is illustrated using a metalloporphyrin template and pyridine as an analyte. The equilibrium binding constant for the axial binding of pyridine to zinc tetraphenylporphyrin has been measured using resonance Raman spectroscopy. Although no new peaks are observed and the porphyrin peaks do not shift position, the quantification is made possible by the selective resonance enhancement of the template vibrations. The value for log k was determined by resonance Raman to be 3.65 +/- 0.32, which compares well with previously published values estimated using absorption data. Values for log k were determined for a series of related compounds, the picolines, and these also compare favourably with those previously reported. PMID- 12572808 TI - Chemical rank estimation by noise perturbation in functional principal component analysist. AB - Some kinds of chemical data are not only univariate or multivariate observations of classical statistics, but also functions observed continuously. Such special characters of the data, if being handled efficiently, will certainly improve the predictive accuracy. In this paper, a novel method, named noise perturbation in functional principal component analysis (NPFPCA), was proposed to determine the chemical rank of two-way data. In NPFPCA, after noise addition to the measured data, the smooth eigenvectors can be obtained by functional principal component analysis (FPCA). The eigenvectors representing noise are sensitive to the perturbation, on the other hand, those representing chemical components are not. Therefore, by comparing the difference of eigenvectors obtained by FPCA with noise perturbation and by traditional principal component analysis (PCA), the chemical rank of the system can be achieved accurately. Several simulated and real chemical data sets were analyzed to demonstrate the efficiency of the proposed method. PMID- 12572809 TI - Fourier-transform Raman characterization of brazilwood trees and substitutes. AB - In this work we have applied Fourier-transform Raman spectroscopy to the analysis of several archival samples of brazilwoods from different geographical origins and of different ages. Samples of Caesalpinia echinata Lam. (from Brazil, South America), Caesalpinia sappan L. (East Indies, Asia), Haematoxylum brasiletto Karsten (Central America) and Haematoxylum campechianum L. (North America) were analysed in order to isolate key Raman biomarker bands which could provide the basis for an identification protocol. Previously recorded Raman spectra of brazilin and brazilein pigments extracted from genuine brazilwood of Brazilian origin provided a foundation for the nondestructive spectral discrimination between a sample of false 'brazilwood', which consisted of an inferior wood substratum stained with genuine pigment, and the true specimens. The provision of well-documented specimens of determinable age from the archival collection facilitated the evaluation of the effects of temporal degradation on the observed spectra, which could be used to further test the experimental protocols for nondestructive verification of samples in the archive with questionable assignment or provenance. PMID- 12572810 TI - Analysis of hydroponic fertilizer matrixes for perchlorate: comparison of analytical techniques. AB - Seven retail hydroponic nitrate fertilizer products, two liquid and five solid, were comparatively analyzed for the perchlorate anion (ClO4-) by ion chromatography (IC) with suppressed conductivity detection, complexation electrospray ionization mass spectrometry (cESI-MS), normal Raman spectroscopy, and infrared spectroscopy using an attenuated total reflectance crystal (ATR FTIR) coated with a thin film of an organometallic ion-exchange compound. Three of the five solid products were found by all techniques to contain perchlorate at the level of approximately 100-350 mg kg(-1). The remaining products did not contain perchlorate above the detection level of any of the techniques. Comparative analysis using several analytical techniques that depend on different properties of perchlorate allow for a high degree of certainty in both the qualitative and quantitative determinations. This proved particularly useful for these samples, due to the complexity of the matrix. Analyses of this type, including multiple spectroscopic confirmations, may also be useful for other complicated matrixes (e.g., biological samples) or in forensic/regulatory frameworks where data are likely to be challenged. While the source of perchlorate in these hydroponic products is not known, the perchlorate-to-nitrate concentration ratio (w/w) in the aqueous extracts is generally consistent with the historical weight percent of water soluble components in caliche, a nitrate bearing ore found predominantly in Chile. This ore, which is the only well established natural source of perchlorate, is mined and used, albeit minimally, as a nitrogen source in some fertilizer products. PMID- 12572811 TI - Batch process monitoring using on-line MIR spectroscopy. AB - Many high quality products are produced in a batch wise manner. One of the characteristics of a batch process is the recipe driven nature. By repeating the recipe in an identical manner a desired end-product is obtained. However, in spite of repeating the recipe in an identical manner, process differences occur. These differences can be caused by a change of feed stock supplier or impurities in the process. Because of this, differences might occur in the end-product quality or unsafe process situations arise. Therefore, the need to monitor an industrial batch process exists. An industrial process is usually monitored by process measurements such as pressures and temperatures. Nowadays, due to technical developments, spectroscopy is more and more used for process monitoring. Spectroscopic measurements have the advantage of giving a direct chemical insight in the process. Multivariate statistical process control (MSPC) is a statistical way of monitoring the behaviour of a process. Combining spectroscopic measurements with MSPC will notice process perturbations or process deviations from normal operating conditions in a very simple manner. In the following an application is given of batch process monitoring. It is shown how a calibration model is developed and used with the principles of MSPC. Statistical control charts are developed and used to detect batches with a process upset. PMID- 12572812 TI - Monitoring solid phase synthesis reactions with electrochemical impedance spectroscopy (EIS). AB - This work describes the use of electrochemical impedance spectroscopy (EIS) as a means to monitor solid phase synthesis on resin beads. EIS was used to track changes during the swelling of beads in various solvents, during three typical reactions and throughout cleavage of the final product from the bead. The impedance response was investigated in a chemical reactor and was found to be faintly sensitive to the resin swelling and solvent flow. The position of the electrode within the reactor was found to be critical as polystyrene based beads float or sink dependent upon the solvent used. However, by choosing electrode position it was possible to monitor reaction progress on beads or within the bulk reactant/product mixture. Of the three typical chemical reactions studied impedance spectroscopy successfully followed two. Fitting of the impedance data to an equivalent electrical circuit provided an estimate as to the relative contribution of capacitive and resistive components to the overall response. Kinetic data from two reactions were also modelled, in both cases complex kinetics was observed, in close agreement with other studies. PMID- 12572813 TI - Optimisation of extraction procedures for metallothionein-isoforms and superoxide dismutase from liver samples using spiking experiments. AB - The speciation of trace element species in solid matrices like liver samples is still problematic due to two reasons. On the one hand direct methods with sufficient selectivity and sensitivity are currently not available. Therefore extraction procedures have to be applied which are often problematic in respect to species stability. On the other hand there are no reference materials with known amounts of metal proteins like metallothionein-isoforms (MT) and superoxide dismutase (SOD) for quality control. So the aim of this study was to develop and optimise procedures for the species-preserving extraction of the model compounds MT and SOD from liver samples. Spiking experiments were performed to overcome the lack of appropriate reference materials. In a first step the stability of the model species without liver matrix was investigated by the variation of several extraction parameters. The extractant and exposure to ultrasonic energy especially had a great influence on the recovery of the species while temperature, buffer concentration and atmospheric conditions were less critical. In a second step spiked liver samples were extracted with a selection of procedures taken from the literature. Most of these methods provided recoveries between 70% and 100%. Additionally the buffer concentration and the extractant-to liver ratio were varied for optimisation. The metal balance of an extraction showed recoveries of 81% for Cd, 94% for Cu and 87% for Zn. PMID- 12572814 TI - Sida's involvement in and support to research in the Western Indian Ocean. PMID- 12572815 TI - Marine protected areas in the eastern African Region: how successful are they? AB - This article reviews the governance and management of Marine Protected Areas (MPAs), and the coral reefs they contain, in the eastern African Region. This includes the Comoros, Kenya, Madagascar, Mauritius, Mozambique, Tanzania, and the Seychelles. Three generations or categories of MPAs are distinguished: i) small areas for protection of a single species or unique marine habitat; iii) large multiple use MPAs designed for coastal development as well as biodiversity protection; and iii) MPAs managed by a nongovernmental organization (NGO) or the private sector. Each of these MPA types is examined according to the policies, legislation, and management systems they entail as well as the economic and community situation they operate within. The paper also provides a review of some eastern African MPAs in terms of their size and location, the type of MPA, zonation schemes, and financial status. The successes of the different types of MPAs are discussed based on specific indicators, such as changes in biodiversity, infrastructure, compliance to regulations and the level of involvement of primary stakeholders in the management. From the review it is clear that a fourth generation of MPAs may be forthcoming; community-based MPAs. Although lack of data makes it difficult to assess the effectiveness of these different categories of MPAs, it is clear that no MPA can succeed without support of the local communities. Generally, the results of the analysis are promising for MPAs, however a lack of data is hampering a deeper analysis. The major issues facing MPAs in the region are highlighted, as well as some regional initiatives striving to address these issues. A number of recommendations are made, aiming to strengthen the establishment and management of MPAs in the eastern African region. PMID- 12572816 TI - Coastal resources and management systems influenced by conflict and migration: Mecufi, Mozambique. AB - Coastal resource utilization and management systems, both traditional and more recently conceived, were studied in Mecufi district, northern Mozambique in a post-conflict situation prior to which a significant migration of people to the coast had occurred. A wide variety of coastal biotopes containing a multitude of resources had been affected in various ways. Intertidal organisms exhibited signs of decreasing abundance and average size, whereas offshore fishes and mangrove forests did not show signs of over-utilization. It was observed that traditional coastal management systems were still influential, but that newer initiatives were only beginning to enter into significant dialogue and cooperation with these. In the current circumstances of peace and political stability, the principal threat to coastal management and the interests of local people in Mecufi is considered to be potential loss of common property resources and land tenure in the face of prospects of privatization, but Mozambican authorities are presently addressing these issues with legal reforms. PMID- 12572817 TI - Marine fisheries in Tanzania. AB - Fishery resources are a vital source of food and make valuable economic contributions to the local communities involved in fishery activities along the 850 km stretch of the Tanzania coastline and numerous islands. Small-scale artisanal fishery accounts for the majority of fish catch produced by more than 43 000 fishermen in the country, mainly operating in shallow waters within the continental shelf, using traditional fishing vessels including small boats, dhows, canoes, outrigger canoes and dinghys. Various fishing techniques are applied using uncomplicated passive fishing gears such as basket traps, fence traps, nets as well as different hook and line techniques. Species composition and size of the fish varies with gear type and location. More than 500 species of fish are utilized for food with reef fishes being the most important category including emperors, snappers, sweetlips, parrotfish, surgeonfish, rabbitfish, groupers and goatfish. Most of the fish products are used for subsistence purposes. However, some are exported. Destructive fishing methods such as drag nets and dynamite fishing pose a serious problem as they destroy important habitats for fish and other organisms, and there is a long-term trend of overharvested fishery resources. However, fishing pressure varies within the country as fishery resources are utilized in a sustainable manner in some areas. For this report more than 340 references about Tanzanian fishery and fish ecology were covered. There are many gaps in terms of information needed for successful fishery management regarding both basic and applied research. Most research results have been presented as grey literature (57%) with limited distribution; only one-fifth were scientific publications in international journals. PMID- 12572818 TI - The artisanal fishery for Octopus cyanea Gray in Tanzania. AB - Preliminary results for the artisanal fisheries of Octopus cyanea Gray (1849) in Tanzania are provided for the period April 2000 until June 2001. A total of 2546 individual catches and 15473 specimens were analyzed from 3 sites located at Tanga, Mafia Island, and Mtwara. Size range, average weight and catch per unit effort (CPUE) were all significantly lower at Tanga and Mtwara compared to Mafia indicating that the former sites may be overfished. Abundance of smaller individuals was higher at Tanga and Mtwara, but overall biomass was lower. Octopi at each site exhibited allometric growth as indicated by analyses of the length weight relationships. Females become sexually mature at a minimum weight of 600 g while for males the minimum weight was 320 g. Higher numbers of mature individuals were found in June of both years and correlate with peaks in the gonosomatic index. Recruitment peaked a few months after brooding periods. Sex ratios indicate females may be more prone to capture during brooding periods. Reasons for differences between sites are discussed. PMID- 12572819 TI - Coastal aquaculture development in eastern Africa and the Western Indian Ocean: prospects and problems for food security and local economies. AB - This paper reviews the experience and status of coastal aquaculture of seaweeds, mollusks, fish and crustaceans in eastern Africa and the islands of the western Indian Ocean. In many respects, coastal aquaculture is still in its infancy in the region, and there is a pressing need to formulate development strategies aimed at improving the income and assuring the availability of affordable protein to coastal communities. This paper also draws from positive and negative experiences in other parts of the world. The requirements of feed and fry, and the conversion of mangroves are used to illustrate how some aquaculture activities constitute a net loss to global seafood production. The paper presents both general and specific sustainability guidelines based on the acknowledgement of aquaculture as an ecological process. It is concluded that without clear recognition of its dependence on natural ecosystems, the aquaculture industry is unlikely to develop to its full potential in the region. PMID- 12572820 TI - Effects of the 1998 coral morality event on Kenyan coral reefs and fisheries. AB - Data were collected in southern Kenya on coral reef ecosystems and fisheries to assess the influence of the 1998 coral bleaching and mortality event. We compared benthic cover, sea urchin and fish abundance in unfished marine parks and fished reefs and the reef-associated fisheries 3 years before and after 1998. Hard and soft coral decreased while coralline algae increased in both management areas. Turf increased in marine parks and sponge and fleshy algae increased in the fished reefs. Sea urchin grazer biomass was unchanged over this period and the fish community changed less than benthic cover. In general, butterflyfish, damselfish and wrasses were negatively influenced while surgeonfish and a few uncommon families were positively influenced by the substratum change. There was a 17% increase in fishing effort as measured by fishermen per day at each landing site and the total demersal catch declined by 8% and the catch per man declined by 21% after 1998. The decline in the total catch and CPUE combined with the increase in effort suggest an overexploited fishery and this makes it difficult to distinguish changes caused by coral mortality or fishing effort. The price of fish increased over this period and this caused an 18% increase in the total value of the fishery but no difference in the net income of individual fishermen. PMID- 12572821 TI - Coral recruitment following a mass mortality event. AB - Coral recruitment, following the coral bleaching episode in 1998, was studied on the Kenyan coast. Scleractinian recruits representing 31 genera from 13 families were recorded in 2001, the dominant families at all sites being the Pocilloporidae, Poritidae, and Faviidae. The highest diversity and density of scleractinian recruits was observed at a site located within a marine park, with 11 families, 20 genera and 21.4 recruits per m2, while an unprotected site and sites with higher sediment input showed recruit densities under five. Survivorship was generally higher in massive than in branching genera. The diversity in the recruit population has decreased compared to pre-bleaching levels, while no significant change in density was recorded. Growth rates in recruits were consistent between sites and time of year, with an average growth rate of 0.117 mm2 mm(-2) mon(-1), with variation between species. The fastest growing genera were Echinopora, Acropora, Pocillopora, and Porites. With the exception of Montipora, the growth rate of surviving pre-bleaching colonies was lower and exhibited greater variability between taxa and sites than among recruits. PMID- 12572822 TI - Environmental effects on the distribution of corallimorpharians in Tanzania. AB - This study examined the distribution and abundance of corallimorpharians (Cnidaria, Anthozoa) in Tanzania in relation to different aspects of the coral reef environment. Five reefs under varying degrees of human disturbance were investigated using the line intercept transect and point technique. Corallimorpharian growth and the composition of the substratum were quantified in different habitats within reefs: the inner and middle reef flat, the reef crest, and at the 2 and 4 m depths on the reef slope. Corallimorpharians occurred on all the reefs and 5 species were identified: Rhodactis rhodostoma, R. mussoides, Ricordea yuma, Actinodiscus unguja and A. nummiforme. R. rhodostoma was the dominant corallimorpharian at all sites. Within reefs, they had the highest densities in the shallow habitats. While R. rhodostoma occurred in all habitats, the other corallimorpharian species showed uneven distributions. Corallimorpharians ranked second, after scleractinian coral, in percent living cover. Results from this study suggested that corallimorpharians benefitted from disturbance compared with other sessile organisms. They preferred inhabiting areas with dead coral, rock and rubble whilst live coral was avoided. There was a positive relationship between percent cover of corallimorpharians and water turbidity and they dominated the more disturbed reefs, i.e. reefs that were affected by higher nutrient loads and fishing. PMID- 12572823 TI - Regeneration status of mangrove forests in Mida Creek, Kenya: a compromised or secured future? AB - The structure and regeneration patterns of Mida Creek mangrove vegetation were studied along belt transects at 2 forest sites of Mida Creek (3 degrees 20'S, 40 degrees 00'E): Uyombo and Kirepwe. Based on the species importance values, the dominant mangrove tree species in Mida were Ceriops tagal (Perr.) C. B. Robinson and Rhizophora mucronata Lamk. Tree density varied from 1197 trees ha(-1) at Kirepwe to 1585 trees ha(-1) at Uyombo and mean tree height was higher at the former site compared to the latter. The size-class structure at both localities of Mida showed the presence of more small trees than large ones. Spatial distribution pattern of adults and juveniles varied greatly between sites and they showed a close to uniform pattern (Morisita's Index I0 << 1) for trees, but a tendency to random distribution (I0 = 1) for juveniles. The present paper shows that unmanaged but exploited mangroves do not necessarily disappear, but change qualitatively from locally preferred R. mucronata to the less preferred C. tagal. Whereas the effects of this change on the ecological function of the mangrove cannot be estimated yet, the economical function of the mangrove has evidently weakened. PMID- 12572824 TI - The East African decapod crab Neosarmatium meinerti (de Man) sweeps mangrove floors clean of leaf litter. AB - Leaf litter removal by the abundant mangrove decapod crab Neosarmatium meinerti was studied in series of field and laboratory experiments in East Africa. In the high intertidal Avicennia marina zone crabs buried all leaves placed on the forest floor and consumed on average 67% of them within 2 hrs. High shore crabs in Kenya buried 4 g m(-2) leaf-litter in 1 hr, i.e. approx. twice the daily litter fall. In contrast, in the low shore Sonneratia alba zone, where typical leaf-eating crabs were absent, none of the offered leaves showed signs of herbivory. Leaf choice experiments in the laboratory showed that N. meinerti preferred some species to others. Leaf consumption per gram crab was higher in females than males. The laboratory studies also indicated that crabs could consume substantially more than the average daily litter fall. Video recordings documented frequent fights to gain or retain fallen leaves, suggesting strong competition for leaf litter. Earlier studies indicating that N. meinerti may sweep mangrove forest floors clean of leaf litter are confirmed. In high shore mangroves of East and South Africa where N. meinerti is common, energy flow appears unique: virtually all litter production is retained. PMID- 12572825 TI - Does the wood-borer Sphaeroma terebrans (Crustacea) shape the distribution of the mangrove Rhizophora mucronata? AB - Field surveys were conducted to evaluate the occurrence of the isopod borer Sphaeroma terebrans (Crustacea) in aerial roots (prop roots) of the red mangrove Rhizophora mucronata on several different spatial scales (m to 100 km) in East Africa. In 6 out of 17 sites studied in Kenya and on Zanzibar Island, Tanzania, no signs of the isopods were found. When the isopods were present the frequency of infestation was high. Trees in muddy substrates in the lower intertidal, in particular at fringing channels or the open sea, showed high prevalence and intensity of infestation, with large part of their roots damaged or dead. Trees at the upper range of Rhizophora, in sandy and muddy areas, showed no signs of isopod infestation. This pattern recurred in mangrove forests on large spatial scales and there was no indication that island forests differed from the mainland forests. This indicates that sediment characteristics, vertical height in the tidal zone, and direct exposure to incoming water are the major factors controlling the abundance of S. terebrans. The isopod may play an important role in determining the lower intertidal limits of R. mucronata. Trees with numerous dead or nongrowing roots, as result of Sphaeroma attack, are likely to tumble due to a lack of root support and this is most likely to occur along channels at the lower, muddy intertidal. Tumbled trees were frequently observed along channels in the lower, muddy intertidal, but rarely in the mid or high intertidal. Implications for management of mangrove forests are discussed. PMID- 12572826 TI - Dynamics of suspended sediment exchange and transport in a degraded mangrove creek in Kenya. AB - This study focuses on sediment exchange dynamics in Mwache Creek, a shallow tidal mangrove wetland in Kenya. The surface area of the creek is 17 km2 at high water spring. The creek experiences semidiurnal tides with tidal ranges of 3.2 m and 1.4 m during spring and neap tides, respectively. The creek is ebb dominant in the frontwater zone main channel and is flood dominant in the backwater zone main channel. During rainy season, the creek receives freshwater and terrigenous sediments from the seasonal Mwache River. Heavy supply of terrigenous sediments during the El Nino of 1997-1998 led to the huge deposition of sediments (10(60 tonnes) in the wetland that caused massive destruction of the mangrove forest in the upper region. In this study, sea level, tidal discharges, tidal current velocities, salinity, total suspended sediment concentrations (TSSC) and particulate organic sediment concentrations (POSC) measured in stations established within the main channel and also within the mangrove forests, were used to determine the dynamics of sediment exchange between the frontwater and backwater zones of the main channel including also the exchange with mangrove forests. The results showed that during wet seasons, the high suspended sediment concentration associated with river discharge and tidal resuspension of fine channel-bed sediment accounts for the inflow of highly turbid water into the degraded mangrove forest. Despite the degradation of the mangrove forest, sediment outflow from the mangrove forest was considerably less than the inflow. This caused a net trapping of sediment in the wetland. The net import of the sediment dominated in spring tide during both wet and dry season and during neap tide in the wet season. However, as compared to heavily vegetated mangrove wetlands, the generally degraded Mwache Creek mangrove wetland sediment trapping efficiency is low as the average is about 30% for the highly degraded backwater zone mangrove forest and 65% in the moderately degraded frontwater zone mangrove forest. PMID- 12572827 TI - Seagrass ecosystems in the Western Indian Ocean. AB - Seagrasses are marine angiosperms widely distributed in both tropical and temperate coastal waters creating one of the most productive aquatic ecosystems on earth. In the Western Indian Ocean (WIO) region, with its 13 reported seagrass species, these ecosystems cover wide areas of near-shore soft bottoms through the 12 000 km coastline. Seagrass beds are found intertidally as well as subtidally, sometimes down to about 40 m, and do often occur in close connection to coral reefs and mangroves. Due to the high primary production and a complex habitat structure, seagrass beds support a variety of benthic, demersal and pelagic organisms. Many fish and shellfish species, including those of commercial interest, are attracted to seagrass habitats for foraging and shelter, especially during their juvenile life stages. Examples of abundant and widespread fish species associated to seagrass beds in the WIO belong to the families Apogonidae, Blenniidae, Centriscidae, Gerreidae, Gobiidae, Labridae, Lethrinidae Lutjanidae, Monacanthidae, Scaridae, Scorpaenidae, Siganidae, Syngnathidae and Teraponidae. Consequently, seagrass ecosystems in the WIO are valuable resources for fisheries at both local and regional scales. Still, seagrass research in the WIO is scarce compared to other regions and it is mainly focusing on botanic diversity and ecology. This article reviews the research status of seagrass beds in the WIO with particular emphasis on fish and fisheries. Most research on this topic has been conducted along the East African coast, i.e. in Kenya, Tanzania, Mozambique and eastern South Africa, while less research was carried out in Somalia and the Island States of the WIO (Seychelles, Comoros, Reunion (France), Mauritius and Madagascar). Published papers on seagrass fish ecology in the region are few and mainly descriptive. Hence, there is a need of more scientific knowledge in the form of describing patterns and processes through both field and experimental work. Quantitative seagrass fish community studies in the WIO such as the case study presented in this paper are negligible, but necessitated for the perspective of fisheries management. It is also highlighted that the pressure on seagrass beds in the region is increasing due to growing coastal populations and human disturbance from e.g. pollution, eutrophication, sedimentation, fishing activities and collection of invertebrates, and its effect are little understood. Thus, there is a demand for more research that will generate information useful for sustainable management of seagrass ecosystems in the WIO. PMID- 12572828 TI - Support to marine research for sustainable management of marine and coastal resources in the Western Indian Ocean. PMID- 12572829 TI - Marine ecology of La Reunion: an overview of recent research. PMID- 12572830 TI - Marine biological research in Mozambique: past, present and future. PMID- 12572831 TI - Patterns in the distribution and abundance of hard corals around Zanzibar Island. PMID- 12572832 TI - Biological N2-fixation on mangrove pneumatophores: preliminary observations and perspectives. PMID- 12572833 TI - Sulfate reduction and methanogenesis in sediments of Mtoni mangrove forest, Tanzania. PMID- 12572834 TI - A review of water quality and pollution studies in Tanzania. PMID- 12572835 TI - Angiokeratoma corporis diffusum--Fabry disease: historical review from the original description to the introduction of enzyme replacement therapy. AB - This paper gives a brief historical overview of Fabry disease. First described in 1898 and called angiokeratoma corporis diffusum, a succession of observations and pioneering clinical research over the past 100 years has led to our present understanding of the disease. The major milestones during this time are described, as is the work that resulted in the successful introduction of enzyme replacement therapy. PMID- 12572836 TI - Aetiology and pathogenesis of hypertrophic cardiomyopathy. AB - The term hypertrophic cardiomyopathy is used to describe an autosomal dominant cardiac disorder, characterized by myocyte hypertrophy and disarray, interstitial fibrosis and small vessel disease, with or without macroscopic hypertrophy. More than 100 mutations in ten genes, all encoding sarcomeric proteins, have been identified as responsible for this disease. Mutations in the genes for beta myosin heavy chain, myosin binding protein-C, and cardiac troponin T are the most common. Other genes involved are alpha-tropomyosin, cardiac troponin-I, essential and regulatory light chains, alpha-cardiac actin, titin, and alpha-myosin heavy chain. Some mutations are more frequently associated with a given phenotype, but no particular phenotype is mutation specific; in fact, some mutations exhibit highly variable clinical, electrocardiographic and echocardiographic manifestations. This variability in the phenotypic manifestations is probably due to the influence of environmental factors and/or modifier genes. While the aetiology of hypertrophic cardiomyopathy has been extensively elucidated, its pathogenesis is not completely understood. Mutated proteins are incorporated in the sarcomere and impair myocyte contractility. This probably triggers the compensatory local release of trophic factors, which influence the development of the typical anatomical features of the disease, with a pathway similar to that observed in secondary, pressure overload hypertrophy. CONCLUSIONS: The various pathological cardiac changes seen in hypertrophic cardiomyopathy are probably due to a compensatory response to impaired myocyte function resulting from mutations in the genes encoding sarcomeric proteins. PMID- 12572837 TI - Cardiac involvement in Fabry disease. AB - Fabry disease is a rare X-linked defect of the lysosomal enzyme alpha galactosidase A. The disease is characterized by progressive intracellular accumulation of neutral glycosphingolipids. The storage occurs within various tissues and cells, including cardiocytes, the cardiac conduction system, and valvular fibrocytes. Cardiac involvement may be the sole manifestation of the disease, particularly in individuals with residual enzyme activity. In general, hemizygous men are more seriously affected than heterozygous women. The main cardiac manifestations include myocardial hypertrophy, which, in some patients, mimics hypertrophic cardiomyopathy. Conduction system involvement leads to PR shortening or, in later stages, to AV blocks. Arrhythmias presenting with variable severity also appear to be common. Valvular involvement is frequently noted but generally mild and clinically non-significant. Newly available enzyme replacement therapy has produced promising results in preventing further functional deterioration of affected organs and possibly also in reversing impaired function. CONCLUSIONS: With the advent of effective enzyme replacement therapy, early diagnosis of Fabry disease may be crucial for patient prognosis. PMID- 12572838 TI - Electrocardiographic signs of hypertrophy in fabry disease-associated hypertrophic cardiomyopathy. AB - AIMS: Fabry disease results from deficient activity of the lysosomal enzyme alpha galactosidase A. Progressive accumulation of the major substrates leads, in both men and women, to progressive hypertrophic cardiomyopathy. We aimed to evaluate the utility of different electrocardiographic (ECG) parameters for assessing the degree and severity of hypertrophic cardiomyopathy in patients with Fabry disease. METHODS: A total of 166 ECGs of 94 hemi- and heterozygous patients with Fabry disease were analysed and compared with echocardiographic-estimated left ventricular mass (LVM). RESULTS: There was a significant (p < 0.0001) correlation between QRS duration (R2 = 0.59), 12-lead amplitude/duration product (R2 = 0.61), Sokolow-Lyon voltage/duration product (R2 = 0.52) and LVM. Analysis of receiver operating characteristics revealed that the 12-lead amplitude/duration product had the highest sensitivity-specificity relationship (p < 0.01 compared with the Cornell index). CONCLUSION: In general, ECG signs of left ventricular hypertrophy correlated well with LVM as revealed by echocardiography. Of the parameters studied, the 12-lead amplitude/duration product was the most successful at describing the severity of cardiac involvement in Fabry disease. These data suggest that ECG parameters have potential for use as a simple and cost-effective means of screening for hypertrophic cardiomyopathy in patients with Fabry disease. PMID- 12572839 TI - Isolated cardiac manifestations in Fabry disease: the UK experience. AB - Cardiac abnormalities are common in patients with Fabry disease, and may be the only clinical manifestation of the disease in some patients. At St George's Hospital Medical School, a national referral centre for hypertrophic cardiomyopathy in the UK, a study of 153 consecutively referred male patients revealed that 4% had Fabry disease. This increased to 6% in patients over 40 years of age. All these patients had electrocardiographic (ECG) abnormalities, most of which were consistent with those found in patients with classic Fabry disease: left ventricular hypertrophy, repolarization abnormalities and a prolonged QRS complex. A study of heterozygote females with Fabry disease also showed ECG and echocardiographic abnormalities in most patients. CONCLUSIONS: Fabry disease should be considered in the differential diagnosis of otherwise unexplained cardiac disease. PMID- 12572840 TI - Mechanisms of neuropathic pain and their importance in Fabry disease. AB - One of the most prominent features of Fabry disease is neuropathic pain. Neuropathic pain occurs after neuronal damage. In contrast to inflammatory or trauma-related pain, which normally helps to maintain or restore body functions, neuropathic pain tends to become chronic, and must therefore be considered a 'pathological' pain. Neuropathic pain has usually been classified according to the aetiology of nerve damage: traumatic, inflammatory, cancer-related or metabolic (e.g. Fabry disease). However, use of this classification often results in inadequate therapy for neuropathic pain. Recent research has revealed distinct mechanisms that are responsible for neuropathic pain. These mechanisms are independent of the aetiology of nerve damage. The most important mechanisms are accumulation and maldistribution of sodium channels on injured axons, pathological sympatho-afferent coupling, disinhibition of nociception and central or peripheral nociceptive sensitization. CONCLUSIONS: Future research should focus on diagnostic tools to identify the predominant mechanisms in individual patients. These mechanism could be targeted specifically by drugs, or non-drug therapy, enabling more effective treatment of neuropathic pain. PMID- 12572841 TI - Evaluation of peripheral and autonomic nerve function in Fabry disease. AB - The neurological manifestations of Fabry disease include severe episodes of lancinating pain and burning paraesthesias in the extremities, often triggered by changes in temperature. The preferential involvement of small nerve fibres and the accumulation of storage product in the central autonomic nervous system and autonomic ganglia means that standard neurophysiological procedures cannot adequately evaluate the peripheral and autonomic nervous systems of affected patients. This paper describes the various methods that have been developed to assess impairment of temperature perception, vibratory perception, sudomotor and sweat gland function, and limb and superficial skin blood flow and vasoreactivity. These methods, including thermal provocation tests, quantitative sudomotor axon reflex testing and venous occlussion plethsmography, have been used effectively in patients with Fabry disease to measure the extent of neurological dysfunction. CONCLUSIONS: Effective methods for measuring neurological involvement in patients with Fabry disease have been developed. These methods will be valuable in assessing the response of patients to enzyme replacement therapy. PMID- 12572842 TI - Pain assessment: the advantages of using pain scales in lysosomal storage diseases. AB - Routine and standardized assessment of pain should be conducted in patients with conditions, such as Fabry disease, that are associated with chronic pain. Such pain assessments, using validated and reliable pain scales or questionnaires, should cover the severity, location, temporal pattern and quality of the pain and how the pain impacts on quality of life and normal daily activity. The severity or intensity of pain can be assessed on verbal descriptor scales, visual analogue scales and numerical rating scales, which rate pain on a scale from 'no pain' through to 'excruciating pain' or pain as bad as you can imagine'. Three pain questionnaires that include such rating scales are short enough to be used repeatedly in a clinical or research setting: the Memorial Pain Assessment Card, the McGill Pain Questionnaire and the Brief Pain Inventory (BPI). The BPI also measures the effect of pain on daily activity and quality of life, defines the location of pain and assesses the effectiveness of previous pain relief medication. CONCLUSIONS: Reliable instruments are available to assess pain in chronic disease. In Fabry disease, these should be used routinely to aid decisions concerning analgesic/pain control medication and to assess the effect of enzyme replacement therapy. PMID- 12572843 TI - Pathophysiology and assessment of neuropathic pain in Fabry disease. AB - Severe neuropathic pain and hypohidrosis are important symptoms of Fabry disease, particularly in the first three decades of life. The pain is associated with a length-dependent small-fibre neuropathy that also causes a selective deficiency of cold perception. Cold exposure often accentuates the pain and worsens thermal perception. The hypohidrosis leads to poor exercise and heat tolerance. The mechanisms by which alpha-galactosidase A deficiency causes these physiological abnormalities are poorly understood. The stored glycolipid (globotriaosylceramide) may interfere with the function of cellular membrane proteins, such as ion channels, or may lead to cytotoxicity. The characteristic neuropathic pain can be symptomatically treated with various types of anticonvulsant drugs, such as carbamazepine. Improvement in neuropathic pain as a primary outcome measure has been useful in demonstrating that enzyme replacement therapy is effective in improving pain-related quality of life in Fabry disease. CONCLUSIONS: The dysfunction of the peripheral nervous system is easily assessable and more readily reversible with specific therapy than the destructive processes that occur in organs such as the kidney. In future, therefore, it is likely that neuropathic pain, quantitative sensory testing and hypohidrosis will serve as clinical outcome measures for studies of specific and effective therapies for Fabry disease. PMID- 12572844 TI - Endothelial markers and homocysteine in patients with classic Fabry disease. AB - AIM: Fabry disease is an X-linked inborn error of glycosphingolipid metabolism due to the deficient activity of alpha-galactosidase A, a lysosomal enzyme. It is a multisystem disorder characterized by progressive renal insufficiency, with added morbidity from cardio- and cerebrovascular involvement. The recent availability of genetically engineered enzyme offers an effective targeted treatment approach, but also emphasizes the need for surrogate markers to delineate organ damage and monitor the efficacy of enzyme replacement therapy (ERT). METHODS: Multiple endothelial factors and plasma homocysteine concentrations were investigated in 12 consecutive hemizygous males with classic Fabry disease and 15 controls as part of an exhaustive baseline evaluation prior to ERT. RESULTS: Compared with the controls, plasma concentrations of homocysteine were significantly (p < 0.01) higher in patients with Fabry disease in the absence of chronic renal failure or vitamin deficiency. Plasma concentrations of vascular cell adhesion molecule-1 were also significantly (p < 0.05) higher in the patients, and there was a trend for decreased endothelin-1 levels. No difference was found in serum intercellular adhesion molecule-1, plasma P-selectin, serum E-selectin and plasma thrombomodulin between the patients and controls. CONCLUSIONS: The results do not reveal measurable evidence for endothelial and leukocyte activation that could reliably serve as surrogate markers for routine monitoring of the efficacy of ERT in patients with Fabry disease. While the exact origin and clinical significance of hyperhomocysteinaemia in Fabry disease remains to be studied in a larger cohort of patients carefully monitored for their concurrent medications, especially carbamazepine, we suggest that patients may benefit from folic acid or multivitamin therapy to treat this additional vascular risk factor, when present. PMID- 12572845 TI - Arterial remodelling in Fabry disease. AB - AIM: The enzymatic defect in Fabry disease results in the slow systemic deposition of uncleaved glycosphingolipids in the lysosomes of vascular endothelium and smooth muscle cells, leading to ischaemic strokes, cardiomyopathy and renal failure. Whereas it is known that Fabry disease affects small blood vessels, little is known about its effects on peripheral large arteries. We therefore set out to compare parameters of arterial wall structure and function in a cohort of patients with Fabry disease and an age-matched control group. METHODS: Large artery phenotype was non-invasively investigated in 21 hemizygous patients with Fabry disease and 24 age-matched male controls. Common carotid and radial artery diameter, intima-media thickness (IMT) and distensibility were determined with high-definition echotracking systems and aplanation tonometry. RESULTS: Patients with Fabry disease had a significant twofold increase in radial artery IMT and distensibility, independent of body surface area, age and mean blood pressure. In both groups, older age at the time of examination was significantly associated with larger radial artery IMT. The relationship between age and radial IMT was 2.3-fold higher in patients with Fabry disease than in controls (p < 0.01). Carotid IMT was mildly but significantly increased in patients with Fabry disease (+18%), whereas distensibility was unchanged. CONCLUSION: This study presents evidence of a major increase in arterial wall thickness and distensibility, measurable at the site of a medium-sized artery, in a cohort of patients with classic Fabry disease. PMID- 12572846 TI - Elevated CNS average diffusion constant in Fabry disease. AB - AIMS: Evaluation of the average brain diffusion constant in Fabry disease. INTRODUCTION: Fabry disease is an X-linked recessive lysosomal storage disorder secondary to deficiency of alpha-galactosidase A and resulting in excess tissue globotriaosylceramide, particularly in cerebral blood vessels. This has been associated with cerebral hyperperfusion. Increased tissue perfusion should increase interstitial water by the Starling relationship. This hypothesis was examined by measuring the average CNS diffusion constant (Dav) in patients with Fabry disease using diffusion-weighted magnetic resonance imaging (DWI). METHODS: Axial DWI was performed at b=0 seconds/mm2 and b = 1000 seconds/mm2 (TR (pulse repetition time), 10000; TE (time to echo), 100; FOV (field of view), 22 cm: 3 mm interleaved slices; image matrix, 128 x 128; GE Signa, 1.5T). Eight healthy male volunteers (age range, 21-47 years) and 17 hemizygous patients with Fabry disease (age range, 19-49 years) were examined. Following DWI acquisition, the trace image and the diffusion distribution map were calculated. The diffusion distribution curve was then fitted by a multi-modal Gaussian curve, allowing estimation of Dav. RESULTS: The Dav was 0.743 +/- 0.024 x 10(-5) cm2/second (mean +/- SD) for patients with Fabry disease and 0.726 +/- 0.014 x 10(-5) cm2/second for the control group. Dav was significantly increased in the patients with Fabry disease compared with the controls (p = 0.029) CONCLUSIONS: The elevated Dav indicates increased brain tissue water diffusivity in patients with Fabry disease, a finding consistent with increased extracellular water and increased cerebral blood flow. PMID- 12572847 TI - Enzyme replacement therapy: from concept to clinical practice. AB - In the early 1960s, the first lysosomal storage disease was identified. Since then over 40 such diseases have been reported. The common feature is that enzyme deficiency leads to accumulation of undegraded macromolecules and lysosomal engorgement, resulting in organ dysfunction. Enzyme replacement therapy (ERT) is being developed for many of these disorders. The present paper summarizes the history of the development of ERT, with particular reference to the mucopolysaccharidoses, and specifically, to mucopolysaccharidosis type VII (MPS VII). The rarity of MPS VII has meant that ERT is not yet available for the small number of affected patients, although the study of MPS VII and murine models of the disease have played an important part in the development of treatment for related disorders, including Gaucher disease. CONCLUSION: Much progress has been made in our understanding of lysosomal storage diseases over the past 40 years. This has led to the development of effective ERT for some of the more common storage diseases, such as Fabry disease and Gaucher disease. Treatment is still awaited, however, for many of the other rare disorders in this area, such as MPS VII. PMID- 12572848 TI - Mucopolysaccharidosis type II--genotype/phenotype aspects. AB - Establishing correlations between a patient's genotype and clinical phenotype is based on the assumption that the same clinical consequences will be observed in individuals with the same residual function of a specific metabolic step. In mucopolysaccharidosis type II (MPS II; Hunter disease), patients present with a wide clinical spectrum. Furthermore, current methods for measuring the activity of the deficient enzyme in MPS II--iduronate-2-sulphatase (IDS)--are insufficiently sensitive to differentiate between complete absence of activity and the presence of residual activity. Attempts have therefore been made to establish genotype-phenotype correlations in order to explain the large degree of heterogeneity and to serve as a better guide to prognosis on which to base genetic counselling and treatment options. Using MPS II as an example, this paper illustrates the difficulties and potential advantages of determining genotype phenotype correlations in lysosomal storage diseases. The response of patients with MPS II to allogenic bone marrow transplantation provides some insight into the likely influence of certain genotypes on therapeutic efficacy. CONCLUSIONS: Evaluation of residual activity of IDS in MPS II using gene analysis, expression studies and transcript analysis does not always allow prediction of a patient's phenotype. The variable response to bone marrow transplantation, however, illustrates the potential importance of determining the genotype for selecting the most appropriate therapy for individual patients. PMID- 12572849 TI - Animal models for mucopolysaccharidoses and their clinical relevance. AB - The mucopolysaccharidoses (MPS) are characterized by the accumulation of glycosaminoglycans (GAG) and result from the impaired function of one of 11 enzymes required for normal GAG degradation. MPS II was the first MPS to be defined clinically in humans and is caused by deficient activity of the enzyme iduronate-2-sulphatase. MPS VI was the first MPS recognized in an animal; since then, all but MPS IIIC and IX have been described as naturally occurring in animals or made by knock-out technology. As in humans, all are inherited as autosomal recessive traits, except for MPS II, which is X-linked. Most animal colonies have been established from single related heterozygous animals, making the affected offspring homozygous for the same mutant allele. Importantly, these models have disease pathology that is similar to that seen in humans, making the animals extremely valuable for the investigation of disease pathogenesis and the testing of therapies. Large animal homologues are similar to humans in natural genetic diversity, approaches to therapy and care, and the possibility of evaluating long-term effects of treatment. Therapeutic strategies for MPS include enzyme replacement therapy, heterologous bone marrow transplantation, and somatic cell gene transfer, all of which have been tested in animals with some success. PMID- 12572850 TI - Enzyme replacement therapy in mucopolysaccharidosis type II (Hunter syndrome): a preliminary report. AB - Mucopolysaccharidosis type II (MPS II; Hunter syndrome) is an X-linked disease caused by a deficiency of the enzyme iduronate-2-sulphatase (IDS), which results in the lysosomal accumulation of glycosaminoglycans (GAG). This paper describes a knockout mouse model of MPS II which has been used to assess the effect of enzyme replacement therapy. Therapy with IDS results in a marked decrease in urinary GAGs, as well as reduced GAG accumulation in several tissues. These studies have been used to support the first clinical trial of recombinant IDS in patients with Hunter syndrome. PMID- 12572852 TI - X-chromosome inactivation and human genetic disease. AB - The inactivation of one X-chromosome in females in early development is the process by which the effective dosage of X-linked genes is equalized between XX females and XY males. The mechanism that brings this about is the subject of intense research. The X-linked gene Xist is a key player, which is necessary but not sufficient for the initiation of X-inactivation. It codes for an untranslated RNA that coats the inactive X-chromosome, which takes on properties characteristic of heterochromatin, but how this change in chromatin is brought about remains unknown. Because of X-inactivation, females heterozygous for X linked genes are mixtures of two types of cells and show a variable phenotype. The proportions of the two types of cells can depart from equality due to cell selection either at the tissue or whole organism level. In rare cases, changes in the Xist gene can cause skewing of X-inactivation. A few genes escape from X inactivation either wholly or partially. CONCLUSION: X-chromosome inactivation is a physiological mechanism that equalizes gene-dosage effects on the sex chromosomes. The occurrence of this normal process affects the phenotype seen in females carrying X-linked mutant genes or chromosome anomalies. PMID- 12572851 TI - Non-viral transfer approaches for the gene therapy of mucopolysaccharidosis type II (Hunter syndrome). AB - AIMS: Hunter syndrome is a rare X-linked lysosomal storage disorder caused by the deficiency of the housekeeping enzyme iduronate-2-sulphatase (IDS). Deficiency of IDS causes accumulation of undegraded dermatan and heparan-sulphate in various tissues and organs. Approaches have been proposed for the symptomatic therapy of the disease, including bone marrow transplantation and, very recently, enzyme replacement. To date, gene therapy strategies have considered mainly retroviral and adenoviral transduction of the correct cDNA. In this paper, two non-viral somatic gene therapy approaches are proposed: encapsulated heterologous cells and muscle electro-gene transfer (EGT). METHODS: Hunter primary fibroblasts were co cultured with either cell clones over-expressing the lacking enzyme or with the same incorporated in alginate microcapsules. For EGT, plasmid vector was injected into mouse quadriceps muscle, which was then immediately electro-stimulated. RESULTS: Co-culturing Hunter primary fibroblasts with cells over-expressing IDS resulted in a three- to fourfold increase in fibroblast enzyme activity with respect to control cells. Fibroblast IDS activity was also increased after co culture with encapsulated cells. EGT was able to transduce genes in mouse muscle, resulting in at least a tenfold increase in IDS activity 1-5 weeks after treatment. CONCLUSION: Although preliminary, results from encapsulated heterologous cell clones and muscle EGT encourage further evaluations for possible application to gene therapy for Hunter syndrome. PMID- 12572853 TI - Pietro M. Gullino and angiogenesis. PMID- 12572854 TI - The role of hydrogen peroxide in endothelial proliferative responses. AB - Hydrogen peroxide (H2O2) is a recently recognized second messenger regulating proliferation in mammalian cells. Endothelial cells possess NADPH oxidases, which produce the H202 precursor superoxide (.O2-) in response to receptor-mediated signaling. Multiple physiologic agents have been shown to stimulate endothelial cells to produce .O2-/H2O2, including growth factors, such as vascular endothelial growth factor and transforming growth factor-beta1, and alterations in biomechanical forces, such as shear stress and cyclic strain. Downstream effects of these stimuli can often be inhibited by scavenging H2O2. Low concentrations of H2O2 stimulate proliferation or enhanced survival in a wide variety of cell types. Also, low concentrations of H2O2 stimulate endothelial migration as well as tube formation in an in vitro model of angiogenesis. Although low concentrations of H2O2 have been shown to be involved in numerous signal transduction pathways and to independently stimulate mitogenesis, there has been little information presented on precisely how mammalian cells respond biochemically to these low concentrations of H2O2. Recently a functional proteomics approach has been utilized to identify proteins responsive to low concentrations of H2O2 in human endothelial cells. PMID- 12572855 TI - Hyperlipidemia coincides with reversible growth impairment of cultured human autologous endothelial cells. AB - Patient-related risk factors for the growth of autologous endothelial cells were assessed in a clinical series of 100 consecutive recipients of in vitro endothelialized prosthetic vascular grafts. For all patients, the indication for bypass operation was arteriosclerotic occlusive disease of the distal arteries. Endothelial cells were harvested from a small piece of subdermal vein and cultured in medium containing 20% of autologous serum. Growth was continually monitored. In cultures that failed to grow, the autologous serum supplement to the culture medium was replaced by pooled homologous serum from young healthy donors. The comparison of a multitude of serum parameters between patients whose endothelial cells failed to grow and those showing normal growth revealed a significant difference in serum lipid content: triglycerides: 4.76 +/- 3.36 versus 2.83 +/- 2.28 mmol/L (p = .001); cholesterol: 6.78 +/- 1.69 versus 5.69 +/ 1.32 mmol/L (p = .003); and lipoprotein (a): 35.9 +/- 28.3 versus 22.2 +/- 26.6 mg/dl (p = .04). Following serum exchange with low-lipid pool serum that contained 1.74 mmol/L triglycerides, 4.86 mmol/L cholesterol, 5 mg/dl lipoprotein (a), and 5.79 mmol/L glucose, a remarkable recovery occurred in 85% of these cultures, resulting in fully restored proliferative capacity. As a consequence, population doubling time did not differ between the two groups at any point in time and mass cultures sufficient for confluent graft endothelialization were obtained with hardly any delay. The authors conclude that hyperlipidemia may lead to growth impairment of cultured human endothelial cells. This growth inhibition is reversible if the supplemented autologous serum is replaced by pooled serum with low lipid content. PMID- 12572856 TI - New human microvascular endothelial cell lines with specific adhesion molecules phenotypes. AB - Vascular endothelial cells recognize blood-borne circulating cells and allow them to extravasate in a tissue-specific manner. Because this property determines the selectivity of lymphocyte homing, it is fundamental in physiological as well as pathological processes (inflammation, autoimmune diseases, metastasis). As a tool to assess the molecular basis of endothelium selectivity, microvascular endothelial cell lines of distinct tissue origin were established. Endothelial cells, isolated from lymphoid tissues (lymph nodes and appendix) and from nonlymphoid immune sites--intestine, lung, and skin--were immortalized in vitro. Their general endothelial characteristics, such as the presence of von Willebrand factor (wWf), angiotensin-converting enzyme (ACE), VE-cadherin, and the intracellular E-selectin, were preserved. This article shows that these cell lines display phenotypic characteristics related to their tissue origin. Hence, endothelial cells from lymph nodes expressed peripheral lymph node addressins (PNAds). Endothelial cells from nonlymphoid tissues were ICAM-1 (intercellular adhesion molecule-1) and CD49e positive, whereas P-selectin was not equally distributed among the cell lines. Endothelial cells from mucosal sites reacted with antibody against human MAdCAM-1 (mucosal addressin cell adhesion molecule). In the adhesion test, lymphoid and myeloid cells adhere to endothelial cell lines in a distinct manner. These lines could be useful to study molecular mechanisms involved in tissue-specific cell-cell interaction. PMID- 12572857 TI - Effect of NADPH oxidase inhibition on E-selectin expression induced by concomitant anoxia/reoxygenation and TNF-alpha. AB - The aim of this study was to quantify the expression of E-selectin, intercellular cell adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vascular endothelial cells (HUVECs) exposed to anoxia/reoxygenation (A/R) in the presence or absence of an inflammatory context (0.1 IU/ml tumor necrosis factor-alpha [TNF-alpha]) and to investigate the effects of two different NADPH inhibitors, apocynin and diphenyleneiodonium (DPI), on the expression of the endothelial cell adhesion molecules. Confluent HUVECs were exposed to anoxia for 3 hours (100% N2), followed by a reoxygenation period of 4 hours. TNF-alpha at 0.1 IU/ml was added to the medium either under normoxic conditions for 7 hours (TNF-alpha) or just before the start of anoxia (A/R + TNF-alpha). Levels of E-selectin, VCAM-1, and ICAM-1 were quantified using specific monoclonal antibodies revealed by an alkaline phosphatase-labeled goat F(ab)'2 fragment against mouse IgG antibody and the fluorescent substrate Attophos. Adhesion experiments were also performed using calcein-labeled U937 leukocytes. HUVECs submitted to A/R overexpressed E-selectin but not VCAM-1 or ICAM-1, whereas TNF-alpha at 0.1 IU/ ml increased the expression of all three adhesion molecules. In endothelial cells subjected to A/R in the presence of TNF alpha, a synergistic increase of E-selectin expression and a synergistic adhesion of U937 cells was noted. The NADPH oxidase inhibitors apocynin and DPI both decreased significantly the U937 adhesion and the E-selectin overexpression on HUVECs submitted to A/R, TNF-alpha, or A/R + TNF-alpha. These results suggest that E-selectin expression is implicated in the leukocyte adhesion to HUVECs caused by A/R in the presence or absence of an inflammatory context. NADPH oxidase appears to participate in the E-selectin overexpression on HUVECs subjected either to A/R and/or TNF-alpha, suggesting a major role of this enzyme in the ischemia/reperfusion syndrome. PMID- 12572858 TI - Regulation of the glyoxalase pathway in human brain microvascular endothelium: effects of troglitazone and tertiary butylhydroperoxide. AB - The glyoxalase system, comprised of glyoxalase-I and glyoxalase-II with glutathione as the cofactor, plays an important role in the detoxification of methylglyoxal and other alpha-oxo-aldehydes. Such aldehydes, which increase with hyperglycemia, give rise to advanced glycation end products. The objective of this research was to examine the glyoxalase system in human cerebromicrovascular cells. The hypothesis tested was that this pathway would be regulated by phase 2 enzyme inducers such as t-butylhydroquinone and modulated by the insulin sensitizing drug troglitazone. Human cerebromicrovascular endothelial cells were cultured and exposed to varying concentrations of t-butylhydroquinone or troglitazone. The activity of glyoxylase-I in human endothelial cells was similar to the activity present in hepatocytes. The phase 2 enzyme inducer t butylhydroquinone had no effect on the glyoxalase enzymes activities but significantly increased glutathione levels and glutathione reductase activity, indicating that phase 2 enzyme inducers might promote alpha-oxo-aldehyde scavenging. Troglitazone decreased the activities of glyoxalase-I and -II and decreased glyoxalase-I mRNA. Troglitazone had no effect on glutathione levels or on the activity of glutathione reductase or glutathione peroxidase. We conclude that phase 2 enzyme inducers may promote scavenging of alpha-oxoaldehydes in endothelial cells. PMID- 12572859 TI - Regulation of epithelial cell functions by the intestinal milieu. PMID- 12572860 TI - Antibodies against hepatoma-derived growth factor and mucosal repair in ulcerative colitis. AB - Ulcerative colitis is a chronic inflammatory disease of colonic mucosa in which the pathogenesis of any immunological disorders would likely be related. Various circulating autoantibodies have been reported in patients with ulcerative colitis, although their possible roles in this disease process have not yet been clarified. Autoantibody against hepatoma-derived growth factor (HDGF) was detected at high frequency in the serum of patients with ulcerative colitis, especially in patients with total colitis and left-sided colitis. In pursuit of the possible role of anti-HDGF autoantibody in the pathogenesis, we investigated HDGF expression in the intestinal mucosa by Western blotting and immunohistochemistry and the effects of recombinant proteins and antirecombinant HDGF antibody on the proliferation of the colonic epithelial cell-derived cell line, HT-29. HDGF was expressed in the nucleus of the colonic epithelial cells dominantly in the bottom of the crypts. Recombinant HDGF stimulated the proliferation of HT-29 cells significantly, although its effects were small, about 20% greater than the control at 100 ng/ml. On the other hand, the polyclonal IgG antibody against recombinant HDGF generated by rabbits suppressed their proliferation almost completely at 250 microg/ml. These findings suggest that HDGF plays an important role in epithelial cell renewal of intestinal crypts as a growth and survival factor, and that autoantibody against HDGF may delay mucosal healing and repair by inhibiting the stimulatory effects of HDGF on epithelial cell proliferation, resulting in a chronic process of colonic mucosal injury. PMID- 12572861 TI - Roles of hepatocyte growth factor activator (HGFA) and its inhibitor HAI-1 in the regeneration of injured gastrointestinal mucosa. AB - Hepatocyte growth factor (HGF)/scatter factor (SF) is thought to play an important role in the regeneration of injured gastrointestinal mucosa by promoting the proliferation and migration of epithelial cells. HGF/SF is secreted by stromal cells as an inactive precursor form, and is specifically activated by HGF activator (HGFA) to the active form. HGFA is also produced as a precursor form and activated by thrombin in injured tissues. The activity of HGFA is regulated by two recently identified Kunitz-type serine proteinase inhibitors, namely HGFA inhibitor type 1 (HAI-1) and type 2 (HAI-2). Although the activation of HGF/SF is a critical limiting step in the HGF/SF-induced signaling pathway, little is known about the regulation of HGF/SF activation in injured gastrointestinal mucosa. Immunohistochemically, all these proteins have been detected in gastrointestinal epithelial cells, and HAI-1 was upregulated in regenerative epithelium relative to normal epithelium. During the course of acetic acid-induced murine experimental colitis, HAI-1, but not HAI-2, was indeed upregulated in the recovery phase. In vitro study revealed that HAI-1 is not only an inhibitor, but also a specific cell-surface binding protein, of active HGFA, and acts as a reservoir of this enzyme on the cell surface. Active HGFA/HAI-1 complexes were quickly released from the cell surface by treatment with IL-1beta accompanying significant recovery of HGFA activity in the culture supernatant. These results suggest that HAI-1 is a cell-surface acceptor of activated HGFA in regenerative epithelial cells, and functions on the cell surface to localize the active HGFA that is going to enter the repair process. This concentrated HGFA activity would ensure the efficient pericellular activation of HGF in the injured gastrointestinal mucosa, and promote the proliferation and migration of gastrointestinal epithelial cells. PMID- 12572862 TI - Possible involvement of neutrophil elastase in impaired mucosal repair in patients with ulcerative colitis. AB - BACKGROUND: Little information is available on the relative contribution of peptide growth factors and leukocyte-derived proteinases to the repair processes in inflammatory bowel disease (IBD). We investigated their possible roles in epithelial cell restitution and proliferation in patients with IBD. METHODS: The expression of hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), transforming growth factor-beta (TGF-beta), and neutrophil elastase (NE) was examined in colonic mucosal tissues. The effects of organ culture supernatants of mucosal tissues on epithelial cell restitution and proliferation were analyzed in vitro using an intestinal cell line, IEC-6 cells. RESULTS: Most organ cultures detected the presence of measurable levels of HGF, with a relative paucity of KGF and TGF-beta activity. Greater levels of HGF were obtained in the mucosa involved with IBD, especially in patients with ulcerative colitis (UC). The mucosa involved with UC also showed higher amounts of NE. The supernatants from the mucosa involved with UC possessed a prominent stimulatory effect on the restitution of IEC-6 cells. By contrast, significant suppression beyond baseline levels was observed for the proliferation of IEC-6 cells when they were incubated with recombinant HGF plus the supernatants from the mucosa involved with UC. This suppression was diminished considerably by preincubation of the supernatants with the anti-NE antibody. CONCLUSIONS: HGF produced in the intestinal mucosa may be an important stimulator acting on epithelial cell restitution in patients with IBD. However, NE released in situ may impair mucosal repair through inhibiting epithelial cell proliferation in patients with UC. PMID- 12572863 TI - Intestinal subepithelial myofibroblasts in inflammatory bowel diseases. AB - Colonic subepithelial myofibroblasts (SEMFs) may play a role in the regulation of a number of epithelial cell functions and in the mucosal repair process. In this study, we evaluated the changes in alpha-smooth muscle actin (SMA)- and vimentin positive SEMFs in the inflamed mucosa of inflammatory bowel disease (IBD) patients. Tissue samples were surgically obtained from patients with active ulcerative colitis (UC) (n = 5) and active Crohn's disease (CD) (n = 5). Normal intestinal tissues were also obtained (n = 5). The SMA and vimentin expression was evaluated by standard immunohistochemical procedures. In normal intestinal mucosa, SMA- and vimentin-positive SEMFs were located immediately subjacent to the basement membrane, juxtaposed against the bottom site of the epithelial cells. In the inflamed mucosa of active UC patients, there were relatively more SMA-positive cells compared with normal mucosa. In particular, the increase in SMA-positive cells was greatest at the marginal area of deep ulcers of UC patients. In active CD mucosa, SMA-positive cells were increased in all samples, and a marked increase was observed in two samples. The number of SMA-positive SEMFs was relatively higher in CD mucosa than in UC mucosa. An [3H]thymidine incorporation study demonstrated that platelet-derived growth factor (PDGF)-BB, basic fibroblast growth factor (bFGF), and insulin-like growth factor (IGF)-I significantly increased the uptake of [3H]thymidine into isolated SEMFs. In particular, PDGF had a strong stimulatory effect. We concluded that colonic SEMFs may play an important role in the repair process of IBD. PMID- 12572864 TI - Role of the innate immune system in the development of chronic colitis. AB - Based on Pasteur's work on the microbial nature of fermentation, it was widely believed that the presence of bacteria in the intestine was essential for the life of the host. It has also been known for decades that gut commensal microbes effect the activation and development of the systemic immune system through gut associated lymphoid tissues (GALT). Recent extensive studies have shown that recognition of microbes is mediated by a set of germline-encoded receptors, Toll like receptors (TLRs), in mammals. This article reviews the role of the innate immunity system in the development of GALT and the pathogenesis of inflammatory bowel diseases (IBD). PMID- 12572865 TI - Intestinal drug delivery systems with biodegradable microspheres targeting mucosal immune-regulating cells for chronic inflammatory colitis. AB - We developed two kinds of delivery systems targeting mucosal immune regulating cells with poly (D,L-lactic acid) microspheres containing dexamethasone and dichloromethylene diphosphonate, and gelatine microspheres containing interleukin 10. To estimate the efficacy of these drug delivery systems, we studied the effects on experimental colitis induced by dextran sodium sulfate, 2,4,6 trinitrobenzene sulfonic acid, and interleukin-10-deficient mice. Intestinal administration of these microspheres significantly improved colitis with decreased histological score, myeloperoxidase activity, and nitric oxide production compared with mice treated with free agents. Gene expressions of tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma were down regulated in treated animals. Serum Dx, IL-10 levels, and systemic macrophages were unchanged after treatment. Our findings suggest that local macrophages in the intestine play a critical role in the initiation of chronic colitis in the animal model of inflammatory bowel disease (IBD). Intestinal drug delivery systems with biodegradable microspheres targeting mucosal immune-regulating cells may become a therapeutic approach to human IBD. PMID- 12572866 TI - Antigen-presenting dendritic cells in ulcerative colitis. AB - BACKGROUND: Antigen-presenting dendritic cells are the inducers and regulators of immune responses. Here, we have discussed the phenotype and function of dendritic cells in situ and their alteration during lymphocytapheresis in patients with ulcerative colitis. METHODS: Dendritic cells were enriched from the peripheral blood mononuclear cells by culturing with granulocyte-macrophage colony stimulating factors and interleukin 4 for 8 days. The function of dendritic cells was evaluated in an allogenic mixed leukocyte reaction. Flow cytometry was employed to study the phenotype of dendritic cells. Lymphocytapheresis was done by a continuous flow centrifugation technique using a CS-3000 separator. Immunohistochemical methodology was employed to detect dendritic cells at the colonic mucosa. RESULTS: Peripheral blood dendritic cells had increased functional capacity, and these cells were matured and activated compared to dendritic cells from normal controls. CD83-positive activated and mature dendritic cells were found at the colonic mucosa from patients with ulcerative colitis. Lymphocytapheresis induced decreased function of peripheral blood dendritic cells in patients with ulcerative colitis. Also, the levels of inflammatory cytokines were reduced by lymphocytapheresis. CONCLUSIONS: Increased function of dendritic cells may be related to the inflammatory mucosal milieu found in patients with ulcerative colitis. Depletion of dendritic cells during lymphocytapheresis may downregulate the exacerbated immune response in patients with ulcerative colitis. PMID- 12572867 TI - Anti-IL-6 receptor monoclonal antibody inhibits leukocyte recruitment and promotes T-cell apoptosis in a murine model of Crohn's disease. AB - BACKGROUND: To assess the contribution of IL-6 signaling to the physiopathology of Crohn's disease, we introduced anti-IL-6 receptor monoclonal antibody to a murine colitis model. METHODS: Colitis was induced in C.B-17-scid mice to which were transferred CD45RBhigh CD4+ T cells from Balb/c mice. Anti-IL-6 receptor monoclonal antibody or rat IgG was given intraperitoneally after T-cell transfer, followed by weekly injection. Vascular adhesion molecules and inducible nitric oxide synthase were visualized by immunostaining. Cytokine expression was determined by RT-PCR, and apoptotic cells were determined by the TUNEL method. RESULTS: Mice treated with anti-IL-6 receptor monoclonal antibody showed normal growth while controls lost weight. Colitis was improved histologically with reduced infiltration of LFA-1+ monocytes/macrophages and VLA-4+ T cells. ICAM-1 and VCAM-1 expression in the colonic vascular endothelium was markedly suppressed by the treatment, whereas no significant difference was seen in MAdCAM-1. IFN gamma, TNF-alpha, and IL-1beta mRNAs were markedly reduced, but no difference was observed in the expression of IL-4, IL-10, and TGF-beta. Inducible nitric oxide synthase was upregulated in the mucosa of colitic mice and downregulated in the treated mice. Apoptotic cells were very sparse despite massive CD4+ T-cell infiltration in colitic mice, whereas increased apoptosis was seen in the treated mice with an apparently reduced number of T cells. CONCLUSIONS: Anti-IL-6 receptor monoclonal antibody abrogated murine colitis. It effectively blocked the expression of adhesion molecules, thereby blocking leukocyte recruitment, and increased T-cell apoptosis. These results strongly suggest the therapeutic potential of anti-IL-6 receptor monoclonal antibody for human Crohn's disease. PMID- 12572868 TI - Gene expression profile after peroxisome proliferator activator receptor-gamma ligand administration in dextran sodium sulfate mice. AB - BACKGROUND: Peroxisome proliferator activator receptor-gamma (PPARgamma) is a member of the nuclear receptor superfamily. Ligands of PPARgamma, thiazolidione derivatives, have been reported to be the one of the candidates for the treatment of inflammatory bowel disease (IBD). Given the fact that PPARgamma is a transcription regulator, expression pharmacogenomics, including differential gene expression profiling of drug responses in a colitis model, is thought to be a useful approach for finding relevant genes that can serve as the target for new drug treatment of IBD. METHODS: We performed a global analysis for differential gene expression of the intestine in a dextran sodium sulfate (DSS) colitis mouse model following PPARgamma ligand administration. By applying a high-density oligonucleotide array method, the expression patterns of approximately 12000 genes were analyzed, and selected genes were confirmed by a real-time quantitative PCR method. RESULTS: The analysis of downregulated genes in the DSS mice following PPARgamma administration revealed several functional gene clusters with altered expression: (1) oncogene families such as GRO1 oncogenes, (2) inflammatory mediator-related genes such as the interferon-gamma gene, (3) water electrolyte-associated genes, and (4) others. CONCLUSIONS: This is the first demonstration of global gene expression analysis using the DSS colitis mouse model with a PPARgamma ligand, and these results provide new insight for finding novel target genes for treating IBD. PMID- 12572869 TI - Treatment of ulcerative colitis by feeding with germinated barley foodstuff: first report of a multicenter open control trial. AB - BACKGROUND: Germinated barley foodstuff (GBF) is a prebiotic foodstuff that effectively increases luminal butyrate production by stimulating the growth of protective bacteria. In the first pilot study, GBF has been shown to reduce both clinical activity and mucosal inflammation in ulcerative colitis (UC). The aim of this study was to investigate the efficacy of GBF in the treatment of UC in a multicenter open control trial. METHODS: Eighteen patients with mildly to moderately active UC were divided into two groups using a random allocation protocol. The control group (n = 7) were given a baseline anti-inflammatory therapy for 4 weeks. In the GBF-treated group (n = 11), patients received 20-30 g GBF daily, together with the baseline treatment, for 4 weeks. The response to the treatments was evaluated clinically and endoscopically. Fecal microflora were also analyzed. RESULTS: After 4 weeks of observation, the GBF-treated group showed a significant decrease in clinical activity index scores compared with the control group (P < 0.05). No side effects related to GBF were observed. GBF therapy increased fecal concentrations of Bifidobacterium and Eubacterium limosum. CONCLUSIONS: Oral GBF therapy may have the potency to reduce clinical activity of UC. We believe that these results support the use of GBF administration as a new adjunct therapy for UC. PMID- 12572870 TI - Intestinal microflora as a therapeutic target in inflammatory bowel disease. AB - Although the causes of inflammatory bowel disease (IBD) remain incompletely understood, increasing evidence implicates intestinal microflora in the pathogenesis of these disorders. Alteration of intestinal flora therefore may offer a plausible therapeutic approach. Although recent data support a potential therapeutic role for probiotics and prebiotics in patients with IBD, such treatments need to be further assessed by large, double-blind controlled trials. A better understanding of the intestinal microflora and the mechanisms of their action may help us to develop more effective treatment for IBD. PMID- 12572871 TI - ICOS costimulation in inflammatory bowel disease. AB - For years, medical researchers have striven to develop selective immunotherapies that could specifically ameliorate pathogenic immune responses without immunocompromising the patient. Blockade of many known receptors on T cells can inhibit the initiation of immune responses. However, this approach is problematic in that it is not possible to predict the onset of disease in patients. Current immunotherapies are unsatisfactory for the sporadic exacerbating type of diseases such as multiple sclerosis and inflammatory bowel disease (IBD), because they require either long-term treatment or acute treatment with high-dose immunosuppressants. With regard to this issue, the inducible and inflammatory site-specific molecule, inducible costimulator (ICOS), may be particularly useful as an ideal targeting molecule for the strategy of treatment of human IBD patients. PMID- 12572872 TI - Analysis of gene expression in human colorectal cancer tissues by cDNA array. AB - BACKGROUND: The development and progression of cancer are accompanied by complex changes in patterns of gene expression. The purpose of this study was to clarify the relevance of macroarray analysis of human colorectal cancer tissues. METHODS: Hybridization of cDNA macroarray filters on which 550 genes had been spotted was performed with biotin-labeled cDNA targets that were prepared from mRNA extracted from 20 pairs of colorectal cancer and corresponding noncancerous tissues. Expression of differentially expressed genes was further studied by semiquantitative RT-PCR. RESULTS: Fourteen (2.5%) of the 550 genes were differentially expressed and up- or downregulated in cancer tissues by at least threefold compared with matched noncancerous tissues in 10 or more of the 20 patients. The genes that were upregulated in cancer tissues were associated with transcription, cell cycle, growth factor receptor, cell adhesion, extracellular matrix-degrading enzymes, and angiogenesis, and the downregulated genes were those involved in apoptosis and immune recognition. Semiquantitative RT-PCR analysis of these differentially expressed genes gave results consistent with those by cDNA array analysis. CONCLUSIONS: Although the macroarray used in this study contained only a small number of genes, our results support the feasibility and usefulness of this approach to study variation in gene expression patterns in human colorectal cancer tissues. The results also suggest the possibility of a diagnostic application of cDNA macroarrays in daily clinical settings. PMID- 12572874 TI - Identification of genes associated with sensitivity to 5-fluorouracil and cisplatin in hepatoma cells. AB - BACKGROUND: The prognosis of patients with advanced hepatoma is grim. Although chemotherapy is adapted to such patients, the efficacy is low and the outcome cannot be predicted before therapy. In this study, we aimed to identify genes associated with sensitivity to 5-fluorouracil and cisplatin, drugs widely used in treatment, using gene expression profiles. METHODS: Gene expression was evaluated in eight human hepatoma cell lines using an in-house cDNA microarray including 2300 known genes. The 50% growth inhibitory concentrations (GI50) of 5 fluorouracil and cisplatin were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay and designated as chemosensitivity. Genes with expression ratios associated with GI50 were selected using the permutation test. RESULTS: For 5-fluorouracil and cisplatin, 21 and 40 genes, respectively, were selected. From among the genes associated with 5-fluorouracil and cisplatin, several encoding metabolic enzymes were selected. In addition, several genes involved in the cell cycle and transcription were identified. CONCLUSIONS: We identified genes that may be associated with sensitivity to 5-fluorouracil and cisplatin. A list of these genes may be useful to elucidate how these drugs work on human hepatoma. PMID- 12572873 TI - p53 gene therapy for esophageal cancer. AB - Despite improvement of surgical treatment and application of multimodality therapies to advanced esophageal cancer, the prognosis is extremely poor for patients with unresectable tumors. Based on the genetic background of esophageal cancer, we have developed various gene therapy strategies against human esophageal cancer. In this article, we review molecular events of esophageal cancer and p53 gene therapy approaches for its treatment. First, we analyzed p53 genetic alterations and angiogenesis in esophageal cancer. Second, we tested a p53 recombinant adenoviral vector (Ad5CMV-p53). Significant growth suppression was observed following infection with Ad5CMV-p53 in human esophageal cancer cell lines. This observation suggests that Ad5CMV-p53 may be a potentially effective therapeutic agent for locally advanced esophageal cancer. Promising avenues for investigation include double gene therapy and adjuvant use of gene therapy with radiation therapy. Third, based on recent reports of clinical trials of p53 gene therapy for lung cancer and head and neck cancer, we developed a clinical protocol for p53 gene therapy for unresectable advanced esophageal cancer. This clinical trial was planned to evaluate vector tolerability and efficacy. Up to December 1, 2001, four patients were enrolled in this phase I/II trial. No serious adverse events related to Ad5CMV-p53 have occurred so far in these patients, and the trial has been safely conducted. PMID- 12572875 TI - DNA methylation changes in gastrointestinal disease. AB - DNA methylation of the 5' region of genes is often associated with gene silencing in X-chromosome inactivation and imprinting. Recent studies have indicated that altered DNA methylation plays a role in the inactivation of multiple tumor suppressor genes and DNA repair genes such as p16INK4A and hMLH1. Colorectal adenomas have a relatively high frequency of methylation, and aberrant methylation is an early event during tumorigenesis. In aging patients, even colon epithelium which appears to be normal showed a significant amount of methylation in a subset of the genes. Colon mucosa from patients with inflammatory bowel disease also showed a high level of methylation. DNA methylation can be a specific diagnostic marker in gastrointestinal cancer and inflammatory bowel disease, for which there is no perfect marker for a noninvasive diagnosis. PMID- 12572876 TI - Association of cholecystokinin A receptor gene polymorphism with cholelithiasis and the molecular mechanisms of this polymorphism. AB - BACKGROUND: The etiology of gallstone formation is multifactorial, and genetic factors are involved. The genetic variations of cholecystokinin A receptor (CCK AR) in patients having gallstones and the molecular mechanisms of this polymorhpism were examined. The involvement of CCK-AR in gallstone formation was confirmed using CCK-AR gene knockout mice. METHODS: CCK-AR gene expression was determined by Northern transfer analysis in gallbladders with or without gallstones. Genetic variations were determined by Southern blot and by direct sequencing. Molecular mechanisms in terms of the transcriptional activity and methylation status were examined. Finally, we investigated whether gallstone formation was enhanced in CCK-AR gene knockout mice. RESULTS: The gene expression of CCK-AR was significantly decreased in gallbladders with gallstones compared to those without gallstones. No genetic variations were detected in the coding region, but two sequence variations were detected in the promoter region in gallstone patients. However, no significant differences were found for the promoter activities of polymorphic promoter constructs. In contrast, less methylation in the promoter region was related to substantial expression of the CCK-AR gene. Gallstone formation was enhanced in CCK-AR gene knockout mice. The homozygote (GG/TT) polymorphism of the CCK-AR gene showed a significantly higher percentage of body fat. CONCLUSIONS: Deteriorating gallbladder contractions, possibly induced by alterations in the CCK-AR gene, as well as CCK-AR gene polymorphism, promoted gallstone formation. PMID- 12572877 TI - Polymorphisms of the IL-1beta and IL-1beta-inducible genes in ulcerative colitis. AB - BACKGROUND: Ulcerative colitis (UC) is a chronic disorder of undetermined etiology, but a genetic predisposition to UC is well recognized. Among cytokines induced in UC, interleukin 1 (IL-1) appears to have a central role because of its immunological upregulatory and proinflammatory activities. The aim of this study was to assess whether UC is associated with polymorphisms of the IL-1beta gene and three additional genes inducible with IL-1beta in Japanese subjects. METHODS: A total of 96 patients with UC and 106 ethnically matched controls were genotyped at polymorphic sites in IL-1beta, matrix metalloproteinase 1 (MMP-1), matrix metalloproteinase 3 (MMP-3), and inducible nitric oxide synthase (iNOS) genes, using polymerase chain reaction (PCR)-based methods. RESULTS: There was no significant difference in genotype distributions of IL-1beta, MMP-1, MMP-3, and iNOS genes between controls and UC patients in a Japanese population. Also, no significant association of those polymorphisms with various clinical parameters of the patients was found. However, concerning association of age at onset with clinical factors in UC, the frequency of pancolitis was significantly higher in UC patients with age at onset being less than 30 years than in those more than 30 years of age (P = 0.049). CONCLUSIONS: No association of the IL-1beta and three IL-1beta-inducible gene polymorphisms with UC was observed in a Japanese population. PMID- 12572878 TI - IL18 polymorphism is associated with an increased risk of Crohn's disease. AB - BACKGROUND: The etiology of inflammatory bowel disease, which includes ulcerative colitis and Crohn's disease, has not yet been made clear. However, inflammatory bowel disease is recognized as a multifactorial disease, and innate genetic factors might contribute to the pathogenesis. Cytokine genes are thought to be important in inflammatory bowel disease. Recently, interleukin 18, cloned as a novel proinflammatory cytokine, has been implicated in inflammatory bowel disease, especially Crohn's disease. METHODS: To identify germline mutations in patients with inflammatory bowel disease, the entire coding region of IL18 was examined using a DNA sequencing procedure. RESULTS: No functional mutations were found, but a novel single nucleotide polymorphism (SNP) was identified as TCA/TCC at codon 35. In patients with Crohn's disease, the frequency of TCC allele carriers was significantly higher than in healthy controls (chi2 = 9.35, P = 0.002229, OR = 2.58, 95% CI = 1.39-4.80). Also, the magnitude of the association was more remarkable in females (chi2 = 16.36, P = 0.000052, OR = 8.17, 95% CI = 2.73-24.41). The TCC allele at codon 35 of IL18 may increase the risk for Crohn's disease, especially in females. CONCLUSIONS: IL18 is probably one of several genes that determine susceptibility to Crohn's disease. PMID- 12572880 TI - Importance of medullary thyrotropin-releasing hormone in brain-gut circuits regulating gastric integrity: preclinical studies. AB - Cold restraint stress (CRS) induced vagal-dependent corpus mucosal erosions with hemorrhage in rats. Aggressive factors such as gastric acid, pepsin, and high amplitude contractions have been reported to be responsible for inducing the formation of gastric lesions. Thyrotropin-releasing hormone (TRH) microinjected into the cisterna magna or the dorsal motor nucleus (DMN) of the vagus induced similar vagally mediated gastric injuries as CRS, and both were prevented by immunoneutralization with TRH antibody injected centrally. These findings indicated that TRH action in the DMN may contribute to CRS-induced gastric mucosal lesions. However, either exogenous or endogenous TRH at a subthreshold dose, which did not increase gastric acid secretion, alleviated gastric injury induced by intragastric administration of a strong irritant in rats. Previous studies have shown that the vagus participates in adaptive cytoprotection. Vagotomy or intracisternal injection of TRH-antibody completely abolished the protective effect of a mild irritant pretreatment in rats. The number of c-Fos protein-positive cells in the DMN increased in the process of adaptive cytoprotection. These results suggest that vagal afferent nerves activated by a topical gastric irritant influence DMN activity by releasing endogenous TRH, leading to protection against injury induced by a subsequent strong irritant. The dual vagally mediated action of TRH in the medulla to regulate the gastric mucosa's response to injury reflects the balance between the aggressive (acid, pepsin, motility) and protective (prostaglandin, calcitonin gene-related peptide, nitric oxide) factors recruited by the level of vagal activation. These data indicate a crucial role of medullary TRH and gastric vagal efferent and afferent circuits in the modulation of gastric integrity. PMID- 12572879 TI - Regulation of feeding behavior, gastric emptying, and sympathetic nerve activity to interscapular brown adipose tissue by galanin and enterostatin: the involvement of vagal-central nervous system interactions. AB - Galanin and enterostatin, which are distributed in both the central nervous system and the gastrointestinal tract, regulate the feeding behavior. In the first set of experiments, we investigated the effects of galanin and enterostatin, injected into the third ventricle, on food intake, gastric emptying, and the sympathetic activity of nerves innervating interscapular brown adipose tissue in rats. Galanin dose-dependently increased the intake of a high fat diet after overnight starvation, but it did not affect low-fat diet intake. In contrast, enterostatin suppressed the intake of the high-fat diet, while intake of the low-fat diet was not affected. Galanin significantly and dose dependently suppressed gastric emptying rate. However, gastric emptying showed no response to enterostatin. Galanin produced a dose-dependent suppression of sympathetic firing rate. In rats fed a high-fat diet, the injection of enterostatin showed a dose-dependent increase in firing rate. In contrast, animals fed a chow diet showed almost no response. In the second set of experiments, we investigated the role of the hepatic vagus nerve in modulating the peripheral response to enterostatin in rats. Intraperitoneal (i.p.) enterostatin reduced the intake of a high-fat diet. Immunohistochemical identification indicated that the Fos protein was present in the nucleus tractus solitarius, and parabrachial, paraventricular, and supraoptic nuclei after IP enterostatin. These responses to i.p. enterostatin were blocked by hepatic vagotomy. These results suggest that galanin and enterostatin coordinate to regulate feeding behavior, gastric emptying, and sympathetic activity to interscapular brown adipose tissue via central and peripheral sites of action, one of which was the interaction which was found to exist through the vagal system. PMID- 12572881 TI - Acid-sensing pathways in rat gastrointestinal mucosa. AB - The gastrointestinal mucosa serves as the interface between the luminal contents, including nutrients and injurious substances, and submucosal structures. Secreted gastric acid is one of the principal injurious components of the luminal contents. To be protected against harm from this acid, the epithelium has an "early warning" system that can activate potent defense mechanisms. We studied the mechanisms that defend the epithelium against luminal acid-induced injury, including the regulation of epithelial intracellular pH (pHi), blood flow, and mucus gel secretion in the perfused rat duodenum, and the pathways involved in the activation and regulation of these mechanisms. Physiological concentrations of luminal acid acidified the epithelial cells and increased blood flow (hyperemic response) and mucus gel thickness. The hyperemic response to acid was abolished by inhibitors of the Na+/H+ exchange, vanilloid receptors (VR), calcitonin gene-related peptide (CGRP) receptors, and nitric oxide (NO) synthase, and also by sensory afferent denervation, but not by pretreatment with a nonselective cyclooxygenase (COX) inhibitor. Mucus secretion in response to luminal acid was delayed by an interruption to the capsaicin pathway, which includes VR, capsaicin-sensitive afferent nerves, CGRP, and NO, and was abolished by COX inhibition. These observations support the hypothesis that the capsaicin pathway is an acid-sensing pathway that promotes hyperemia and mucus secretion in response to luminal acid. The COX pathway is a secondary regulatory system for mucus secretion. A similar acid-sensing capsaicin pathway is also present in the colon, suggesting that the gastrointestinal mucosa "tastes" luminal acidity through epithelial-VR communication. PMID- 12572882 TI - Homeostasis in the small intestinal mucosa balanced between cell proliferation and apoptosis is regulated partly by the central nervous system. AB - The aim of this study was to investigate whether the central nervous system regulates mucosal cell growth and apoptosis in the rat small intestine. Ornithine decarboxylase is a key enzyme for polyamine synthesis, which plays an important role in intestinal mucosal growth. The increase in ornithine decarboxylase activity in the duodenum just before a dark period was abolished by truncal vagotomy. An infusion of 2-deoxy-D-glucose into the third cerebroventricle activated the enzyme activity in the small intestine. Epithelial homeostasis is balanced by the regulation of cell proliferation and cell death. Intestinal mucosal apoptosis decreased in rats with ventromedial hypothalamus lesions, which induced hyperphagia and obesity. In contrast, sustained anorexia induced by 1 deoxy-D-glucosamine increased intestinal apoptosis. These results indicate that the central nervous system, in addition to local factors, is related to the regulation of mucosal homeostasis in the intestinal mucosa. PMID- 12572883 TI - Exaggerated motility of the descending colon with repetitive distention of the sigmoid colon in patients with irritable bowel syndrome. AB - BACKGROUND: Visceral hypersensitivity is one of the mechanisms of irritable bowel syndrome (IBS), but it does not explain the entire symptomatology, i.e., altered bowel habit with abdominal pain relieved by defecation. We tested our hypothesis that an abnormal link between luminal stimulation and mural response may have some role in the pathophysiology of IBS. METHODS: Patients with IBS (n = 10, median 21 years old, 5 male patients, 5 female patients) and healthy control subjects (n = 10, median 21 years old, 5 men, 5 women) were studied. A manometric catheter with three transducers was inserted to the descending colon and a balloon was placed in the distal sigmoid colon. Another catheter with three transducers was inserted to the duodenum. After baseline for 30min, the sigmoid colon was stimulated by balloon distention for 30min followed by recovery for 30min. Balloon distention was repeated 100 times, and each stimulation consisted of a 5-s inflation and a 10-s deflation, with a volume of 50ml maximum. The sensory threshold of balloon inflation was then examined, and plasma adrenocorticotropic hormone (ACTH) was measured with radioimmunoassay. RESULTS: Repetitive colonic distention induced a significant increase in motility indices (mmHg s/s%) of the descending colon in the IBS patients (from 118 +/- 25 to 333 +/- 108, P < 0.05) but not of those in controls (from 90 +/- 16 to 89 +/- 19). A significant group difference (P < 0.05), period effect (P < 0.02), and group x period interactions (P < 0.01) were detected with two-way ANOVA. Duodenal motility indices in controls were significantly reduced by colonic distention (from 169 +/- 25 to 104 +/- 14, P < 0.01), but those in the IBS patients were not (from 156 +/- 17 to 124 +/- 20). The sensory threshold of balloon inflation in the IBS patients (74 +/- 10ml) was significantly lower than that in controls (125 +/- 6ml, P < 0.001). There was no significant difference in plasma ACTH levels between the IBS patients and controls. CONCLUSIONS: Repetitive distention of the distal sigmoid colon below the sensory threshold induced orad exaggerated motility of the colon in IBS patients. The distention inhibited motility of the small intestine in healthy subjects, but this inhibition was blunted in IBS patients. These results suggest that IBS patients may have not only visceral hypersensitivity, but also an abnormal intestinal reflex. PMID- 12572884 TI - Brain-gut axis of the liver: the role of central neuropeptides. PMID- 12572885 TI - Participation of hepatoma-derived growth factor in the regulation of fetal hepatocyte proliferation. AB - The identification and characterization of hepatic stem cell compartments is the key to resolving clinical disorders and diseases in the liver. Hepatoblasts (or early fetal hepatocytes) fulfill several criteria of hepatic stem cells during liver development. Unlike mature hepatocytes, immature fetal hepatocytes can proliferate autonomously in the absence of any growth factors in vitro. However, the regulation of fetal hepatocyte proliferation remains unclear. Recently, we identified a novel factor, hepatoma-derived growth factor (HDGF), from the human hepatoma-derived cell line HuH-7, which autonomously proliferate in serum-free defined medium. Here, we focus on the functional roles of HDGF, and review several molecules involved in the growth regulation of hepatocytes in the immature stage. PMID- 12572887 TI - Vegetable lipids as components of functional foods. AB - Nutritionally beneficial compounds naturally present in vegetable lipids will be subject of this minireview. This article will discuss lipidic compounds from less known vegetable sources and potential advantages of its incorporation into human diet as a functional ingredient. PMID- 12572886 TI - A new cell therapy using bone marrow cells to repair damaged liver. AB - Hepatic stem cells can transdifferentiate into hepatocytes, bile duct cells, pancreatic cells, or intestinal cells. Hepatic stem cells had been thought to be located in the canal of Hering in the liver, but recent work has demonstrated the existence of hepatic stem cells in bone marrow as well. Cell therapy using autologous bone marrow cells has few ethical problems and many applications for treating severe liver disease. Further in vitro and in vivo analysis is crucial to develop the therapy for clinical use. PMID- 12572888 TI - Peroxisome proliferator-activated receptors (PPARs) in health and disease. AB - Peroxisome proliferator-activated receptors (PPARs) belong to the superfamily of steroid-thyroid-retinoid nuclear receptors. PPARs are transcription factors activated by specific ligands and play an important role during cell signalling. Intensive study of PPARs during recent years has revealed their importance in both normal physiology and in the pathology of various tissues. They participate in the regulation of lipid metabolism, inflammation and the development of atherosclerosis or diabetes. They also play a role in the regulation of growth and differentiation of cancer. It has been suggested that PPAR ligands may have potent anticancer effects and therefore may serve as potential anticancer drugs. In this review we focus on a role of PPARs in breast cancer and in glial tumors of the brain. PMID- 12572889 TI - Oxidative burst of Kupffer cells: target for liver injury treatment. AB - Liver injury, and frequent consequent fibrosis, is the focus of a number of research groups ranging from molecular biologists to clinicians. It encompasses many aspects and approaches. Among biochemical events the role of Kupffer cells, oxidative stress and pro-inflammatory mediators are eminent. In this review we focus on recent findings into use of natural substances for the modulation of oxidative burst as well as production of inflammatory mediators by Kupffer cells. PMID- 12572890 TI - Particle disease. A comprehensive theory of periprosthetic osteolysis: a review. AB - Aseptic loosening and osteolysis are considered the main long-term problems of hip arthroplasty. Pathogenesis of periprosthetic osteolysis is multifactorial, and both the biological and mechanical factors seem to play an important role. Bearing surfaces continuously generate excessive amounts of micron and submicron particles provoking an adverse inflammatory response of periprosthetic connective tissues. In general, a key role has been attributed to macrophages. Cytokines, growth factors, PGE2, and enzymes are secreted with activated periprosthetic cells resulting in formation of osteolytic granulomas. The final osteolytic step is taken predominantly by osteoclasts which are getting ready for action mainly by an osteoprotegerin ligand (RANKL) and TNFalpha. Rankl is expressed by activated macrophages, osteoblasts, and lymphocytes. In parallel, a repetitive hydraulic effect of the joint fluid is manifested on the susceptible bone. PMID- 12572891 TI - A microbiological approach to acne vulgaris. AB - The present article gives a concise survey of contemporary opinions on acne vulgaris, its etiopathogenesis, clinical forms and laboratory diagnostics. In particular, the value of microbiological diagnostics and possibilities of local as well as general therapy are discussed. Moreover, our experience is described with vaccinotherapy to manage serious clinical forms and cases when current therapy fails. PMID- 12572893 TI - A sugar, laundry detergent, and salt method for extraction of deoxyribonucleic acid from blood. AB - We present a method for DNA extraction from peripheral blood using sugar, commercial detergent, and sodium salt. Our method is simple, fast, and inexpensive; its qualitative parameters do not significantly differ from the standard salting-out procedure. PMID- 12572892 TI - Serodiagnostics of chlamydial infections--significance of positivity in IgA and/or IgM antibody classes only. AB - There was followed the development of serological findings in patients with proved positivity only in classes IgA and/or IgM of chlamydial antibodies (without IgG), which can be suspected of showing "false" positivity. 184 patients were repeatedly examined for chlamydial antibodies in their sera (interval between collections up to three months) using a genus specific rELISA. Sera were also tested for the evidence of IgM antibodies against capside antigen of Epstein Barr virus (EBV) and against cytomegalovirus (CMV) using ELISA methods. In 75 (40.8%) of patients, IgA/IgM individual positivities were demonstrated even during the following sample test(s). In 28 (15.2%) of them, IgG evidence preceded and in 29 (15.7%) other patients positive seroconversion followed in this class. In 13 (7.1%) patients, IgG antibodies disappeared and subsequently reappeared. Only in 39 (21.2%) of these probands, antibodies IgA/IgM were not demonstrated at another examination. Active EBV, resp. CMV infection was proved in 24 (13.0%), resp. in 18 (9.8%) of patients. It is concluded that the evidence of positivities only in classes IgA and/or IgM mostly signal the onset of a primary infection (reinfection) or an active infection in patients with IgG production failures respectively. In these cases, a "false" positivity can be supposed to occur only in a minor extent. PMID- 12572894 TI - Part of CD68+ macrophages in the clearence of apoptotic bodies in human metanephros. AB - According to recent research on mice, less on human material, cells responsible for clearing apoptotic cells away during development are, besides non professional phagocytes, also tissue-fixed macrophages. The aim of our work was the determination of macrophage role in the phagocytosis of apoptotic bodies in neogenous zone of human metanephros. Histologicaly normal kidneys were collected from embryos and fetuses ranging from the 8th-28th week of IUD. These tissues were routinely processed. In the first step we detected CD68+ cells by means of standard indirect three-step immunohistochemical method having used MAb NCL-CD68 KP1 (macrophage marker) to find out whether such cells are actually present. In the second step tissue sections were labelled by double-staining principle (TUNEL technique for the detection of apoptosis and above mentioned macrophage marker) to judge co-localization of these two items. The slides were observed by using immersion objective and the amount of apoptotic cells was expressed in percents. CD68+ macrophages appeared dispersely as single cells or small groups in all the ages studied. According to our results, CD68+ macrophages phagocytose 37-75% of apoptotic cells present in neogenous zone and the number of engulfed apoptotic cells increases in the 12th week of the IUD, i.e. in the early fetal period and later it merely fluctuates. PMID- 12572895 TI - Preliminary experience with resistin assessment in common population. AB - Resistin is a signal peptide produced by adipose tissue. Mice models have confirmed that resistin may play an important role in insulin resistance. Its function in the human organism has not been elucidated yet. Since in common population the resistin concentrations are not known (no validated commercial set is available), we performed resistin assessment using the ELISA method (with satisfying analytical characteristics) in a population of 123 non-obese probands without signs of insulin resistance and/or inflammation. Mean resistin values amounted to 14.3 ng/ml (reference limit of 7.3-21.3 ng/ml). PMID- 12572896 TI - Heterotopic cardiomyocytes in the wall of pulmonary vessels in experimental animals. AB - Heterotopic cardiomyocytes (CMC) have been described in the wall of pulmonary veins including intraparenchymatous branches, in various species of laboratory animals. Brief note is given here of their morphology including electron microscopy and immunohistological findings. From the current literature hints concerning arrhythmogenic activity of heterotopic CMC in pulmonary vessels in the possible initiation of atrial tachyarrhythmia and fibrillation are also mentioned. PMID- 12572897 TI - Contribution to differential diagnostics of granulomatous lesions in lung tissue. AB - Etiopathogenesis of pulmonary granulomatous lesions can be very varied. Pathologists find pulmonary granulomatous lesions in autopsies frequently. Diagnostics of pulmonary granulomatous lesions is based on anamnesis data, clinical examinations and morphological changes. In some cases, especially in long-term ill patients, morphological finding remains the only way to explain the case. We provide a description of an incidental autoptic finding of chronic granulomatous lesion in lung tissue in a man aged 81. Histopathological examination proved aspired granules of maize starch. Aspired granules triggered lesion of granulomatous character. At microscopic examination vegetable particles were defined to be maize starch. Granules of maize starch, which are a part of artificial nutrition labelled Caglusal, may have caused granulomatous lesion in our case. PMID- 12572898 TI - Monitoring of bone resorption and bone formation in multiple myeloma. AB - The article deals with the clinical value of monitoring of serum markers of osteoresorption (ICTP) and bone formation (PICP) in multiple myeloma. In a group of patients treated by conventional chemotherapy and group of patients treated by high dose chemotherapy with autologous peripheral blood stemm cell transplantation (APBSTC). PMID- 12572899 TI - An uncommon severe clinical course of European tick-borne encephalitis. AB - Tick-borne encephalitis (TBE) is the major European arbovirosis. Man is often infected by the tick bite; laboratory infections as well as infections after consumption of nonpasteurized milk have occasionally been reported. TBE typically takes a biphasic course. After an incubation period (7-14 days usually), the prodromal symptoms occur. The first stage of the disease lasts 1-8 days. After an afebrile asymptomatic interval of approximately 1-week the second stage develops, in which TBE may manifest as isolated meningitis, meningoencephalitis, meningoencephalomyelitis and/or radiculitis. The mortality rate is 0-3.3%. There is high frequency of sequelae. Patients who have suffered from a TBE infection are immune for the rest of their lives. At present there is no effective antiviral therapy and management is strictly supportive. Active immunization against TBE is recommended for all subjects living in and travelling to areas of risk. Passive immunization is recommended only within 48 hours after a tick bite. The authors present the case report of 75-years old female subject, who suffered from the uncommon severe clinical course of TBE--with the development of consciousness impairment, tetraplegia, and dyspnoea (demanding ventilatory support). Her state was complicated by the transient left side faciobrachial motor Jackson's convulsions, blood pressure instability, heart arrhythmia, respiratory infection, anemia, and by the development of critical illness neuropathy. Although she made slow progress during treatment, she died on the 114th day because of suspected aspiration. PMID- 12572900 TI - Dialysis activity of the Hemodialysis (HD) Center at University Hospital Olomouc in the past decade. AB - The paper presents a summary of dialysis statistics at the Olomouc Hemodialysis Center during the period 1991-2001. The number of individual treatments, patient structure, mortality and survival of patients on dialysis are featured, along with a comparison of state norms in some instances. PMID- 12572901 TI - Impaired triglyceride tolerance in hemodialysis patients with different apolipoprotein E (apo E) isoforms. PMID- 12572902 TI - Venous hypertension following average arterious-venous fistula for haemodialysis. AB - The authors emphasize the need for the investigation of the central venous system prior to the insertion of an avfistula, this they consider to be of utmost importance in patients with anamnesis of central vessel access. After the av fistula is inserted, an unrecognized obstacle (stenosis or thrombosis) may result in the occurrence of venous hypertension and hypofunction, this may lead to av fistula malfunction. PMID- 12572903 TI - Assessment of spectral analysis of heart rate variability in patients with history of atrial fibrillation by means of age-dependent parameters. AB - Heart rate variability evaluation is a useful diagnostic tool for autonomic nervous balance assessment. The role of the autonomic nervous system in aetiology of atrial fibrillation is sometimes clear as a trigger from a patient's history, but mostly it acts as a modulating factor which is not easy to detect. The present study demonstrates results of spectral analysis of short-term heart rate variability during ortho-clinostatic tests processed by means of age-dependent parameters. An original telemetric system and a unique method for heart rate variability assessment, developed by the Faculty of Physical Culture, were applied for the first time to examine patients with the history of atrial fibrillation. PMID- 12572904 TI - Comparison of the efficacy and safety of Consupren solution and Sandimmun Neoral solution, 50 ml in stable heart transplant patients. AB - Cyclosporine A (CyA) is a standard component of immunosuppressive regimen after heart transplantation in most centres. The widespread clinical use of cyclosporine-based immunosuppressive regimens since 1983 has led to significant improvements in the survival of cardiac allograft recipients due to decreased mortality from infections and rejections. (1-3) CyA has been shown to be safe and effective. Owing to its success when used after the heart transplantation the number of patients has also risen. This caused growing financial demands on health insurance companies in the Czech Republic where the immunosuppressive drugs are fully reimbursed. A prospective randomized study in 11 stable heart transplant patients was performed to compare the efficacy and safety of Consupren (IVAX-CR) a Sandimmun Neoral (Novartis) solution based immunosuppressive regimen. The results suggest that Consupren solution can be used as an alternative treatment to Sandimmun Neoral in CyA based regimen. PMID- 12572905 TI - The spectral analysis of syllables in patients using dentures. AB - Changes in the oral cavity resulting from the loss of teeth and the ensuing reconstruction of a set of teeth by dentures (partial or complete) may cause changes in the speech and voice of the patient. The aim of the present investigation was to study the changes in speech and voice in patients suffering from teeth loss and the degree of speech improvement using dentures. Voice and speech parameters of a set of tested syllables were analysed in 10 patients at the 2nd Clinic of Stomatology. The analysis was carried out by means of an FFT, SoundForge 5.0 programme. Differently expressed acoustic changes in both consonants and vowels were ascertained in a percentage of the patients under examination. These concerned especially the sibilant ("s", "(see text)"), labiodental ("f", "v") and vibrating ("r", "(see text)") consonants. Changes in the FFT spectrum and air leakage in constrictive consonants were also found. In some patients the vowels, especially the closed ones ("i", "u"), may change their fundamental frequency and show noise admixture manifested as a blurred delimitation of the formants. A denture should, inter alia, render it possible for the patient to produce the same articulation to which he/she had been accustomed before the loss of teeth. For the construction of dentures the most important factors from a phonetic point of view appear to be the following: overbite, overjet, the height of the plate, the thickness of the palatal material, the incisor position, and the modelling of the ruga palatina on the hard palate. In case of wrong denture construction the acoustic changes may continue, resulting in the patient's stress load dependent upon sex, age, psychic condition and seriousness of the problem. PMID- 12572906 TI - Contribution to the coincidence of malignant tumours and some allergic manifestations. AB - Clinical observations during the past decades led us to the early empirical assumptions that adult patients affected by certain types of allergic manifestations may have a lower prevalence of malignant tumors. In order to test those observations as well as some contradictory reports (1-3), we conducted a retrospective study using 32 years of records and statistics of a medium-sized hospital (420 beds) at St. Mary's Hospital, a general, non-chronic, teaching hospital, affiliated with McGill University in Montreal. The study has been realized during two periods: (a) 1965-1989 and (b) 1990-1996. PMID- 12572907 TI - Preventive oncologic check-ups in the District Sumperk, 1975-1986: the description of a database. AB - From 1975 through 1986, repeated preventive oncologic check-ups were carried out in the District Sumperk. In the course of the check-ups, 49,426 persons were examined. The examinations were aimed at surveillance of initial stadium of cancers as well as at detection of other serious chronic diseases. The results of the check-ups form a large database suitable for a number of epidemiological analyses. In this article, methods of the data collection are described, individual data items are characterized and their summary is presented in tables. Utilization of the database is proposed in specified types of epidemiological studies. PMID- 12572908 TI - Epidemiology of lung cancer. AB - This review article presents lung cancer epidemiology, describing main epidemiologic characteristics including epidemiological situation in cancer incidence, mortality and survival in Europe in comparison with situation in the Czech Republic. Influence of environmental and life style risk factors like smoking, passive smoking, risk factors of work environment, ionizing radiation, air pollution, nutrition and genetic and hormonal factors are discussed. PMID- 12572909 TI - Trends in Europe to reduce the indoor air pollution of VOCs. AB - A strong research tradition in the European countries about volatile organic compounds (VOCs) and indoor air quality has led to several consensus reports about stategies for VOC measurements including the development of methods to measure VOC emissions from building products. European and Nordic standards have been developed for emission testing including several national and international labeling schemes. Proposals for guidelines for a number of different VOCs have been developed either nationally or by consensus. A compound-by-compound approach for health evaluation of VOCs has become more common and there appears to be a growing recognition to focus on the biologic relevance of organic compounds in indoor air (OCIAs). PMID- 12572910 TI - Organic compounds as indicators of air pollution. AB - The most important indoor air pollutants have already been addressed with individual national guidelines or recommendations. However, an international set of guidelines or recommendations for indoor air quality (IAQ) is needed for these pollutants based on general and uniform rules for setting such standards. A major research need exist on the less adverse pollutants before recommendations or guidelines can be established. In the interim period a precaution principle should lead to an ALARA principle for these secondary causalities. It should be noted that volatile organic compound (VOC) as an IAQ problem still is in the end of a phase of ad hoc solutions, in the middle of a research phase and only in the beginning of a regulatory phase. Any final official regulation in this area will have to be tentative and the final regulation must await further research. Total volatile organic compound (TVOC) is an indicator for the presence of VOC indoors. The TVOC indicator can be used in relation to exposure characterization and source identification but for VOCs only, not as an indictor of other pollutants and their health effects. In risk assessment the TVOC indicator can only be used as a screening tool and only for sensory irritation. PMID- 12572911 TI - Modeling and CFD prediction for diffusion and adsorption within room with various adsorption isotherms. AB - This paper presents physical models that are used for analyzing numerically the transportation of volatile organic compounds (VOCs) from building materials in a room. The models are based on fundamental physicochemical principles of their diffusion and adsorption/desorption (hereafter simply sorption) both in building materials and in room air. The performance of the proposed physical models is examined numerically in a test room with a technique supported by computational fluid dynamics (CFD). Two building materials are used in this study. One is a VOC emitting material for which the emission rate is mainly controlled by the internal diffusion of the material. The other is an adsorptive material that has no VOC source. It affects the room air concentration of VOCs with its sorption process. The floor is covered with an emission material made of polypropylene styrene-butadiene rubber (SBR). An adsorbent material made of coal-based activated carbon is spread over the sidewalls. The results of numerical prediction show that the physical models and their numerical simulations explain well the mechanism of the transportation of VOCs in a room. PMID- 12572912 TI - Performance analysis on hybrid ventilation system for residential buildings using a test house. AB - Hybrid ventilation system is a two-mode system that can automatically switch between passive and mechanical modes at different times of the day or season of the year. In this study various ventilation systems, including hybrid system, are used for the investigation. These systems are set up in a full-scale test house constructed in the Tohoku University, Japan. Case studies for evaluating the performance of these systems, by field studies and numerical simulations, are described. The results obtained from this study show that, the hybrid system can supplement the under-ventilation even under a milder weather condition and the airflow rate can be fixed at a certain high level, in comparison with passive system. PMID- 12572913 TI - Database system, AFoDas/AVODAS, on indoor air organic compounds in Japan. Automated Formaldehyde Data Acquisition System. Automated VOC Data Acquisition System. AB - Building designers, engineers and occupants often need to estimate the organic compounds' levels in buildings. In order to satisfy the requirement, a web-based database, named AFoDAS/AVODAS database system is developed. The database consists of six databases for managing indoor air organic compounds' levels and building's environmental conditions. The database includes the monitoring results of about 1422 homes. The database allows users to (1) input their monitoring results to the database; (2) browse, search and query the database by a single or combinations of criteria for various query parameters such as building category, ventilation system, environment category, and compound names; and (3) estimate the organic compounds' levels resulting from various buildings and environmental conditions. This paper describes the functionality, database structure and major features of the user interface system of the database. PMID- 12572914 TI - Field survey of indoor air quality in detached houses in Niigata Prefecture. AB - In this study, measurements of indoor air quality and ventilation rate of detached wooden houses in Niigata Prefecture are carried out. The air quality (the concentration of formaldehyde (HCHO) and volatile organic compounds (VOCs), the air-tightness of the houses and the life style of the occupants were investigated. The temperature and humidity were logged for 1 week at 10-min intervals using stand-alone data loggers. The measuring points were at 110 and 5 cm above the floor and 10 cm below the ceiling. A 2,4-dinitrophenylhydrazine coated silica (DNPH) cartridge (Waters Sep-Pak XpoSure) was used to measure the concentration of HCHO and a passive air sampler (3M Organic Vapor Monitor) was used to measure the concentration of VOCs in the living room for 1 week. High performance liquid chromatography (HPLC) and gas chromatography with mass spectrometry (GC/MS) were used to analyze the average weekly concentration of VOCs. The weekly averaged ventilation rate of the house was measured by perfluorocarbon tracer gas technique (PFT). The main results can be summarized as follows. 1. A total of 38.2% of the occupants felt that the indoor air quality was bad when cooking or smoking in the house took place. 2. The HCHO concentration in the houses is inversely proportional to the air leakage from the house, i.e. it was high for airtight houses. 3. The HCHO concentration in older houses is lower than in new houses of similar air-tightness. 4. When building age increases, HCHO concentration in indoor air decreases. 5. Generally the concentration of benzene, toluene, p-dichlorobenzene, o-, m-, p-xylene is high, especially in a newly constructed house. PMID- 12572915 TI - The right to healthy indoor air: status by 2002. AB - One of the reasons for the inadequate quality of indoor air arises from the poor articulation, appreciation and understanding of basic principles underlying the policies and actions related to indoor air quality. A WHO Working Group derived nine statements on rights to healthy indoor air. The discussions and statements are available as a WHO report. It informs the individuals and groups responsible for healthy indoor air about their rights and obligations, and empowers the general public by making people familiar with those rights. One year after their publication the statements have been adopted as the base for future regulation and guidance. The Board of Directors of the International Society of Indoor Air Quality (ISIAQ) and the participants of two international conferences endorse the use of the statements. No opposition to the statements have been registered. The statements have entered curricula of training courses and have been used in lawsuits. PMID- 12572916 TI - Molecular genetics of frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). AB - Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) is characterised by behavioural, cognitive and motor disturbances. Pathological changes in the brain include fronto-temporal atrophy with neuronal loss, grey white matter gliosis and superficial cortical spongiform. In addition, intraneuronal tau inclusions with the variable occurrence of glial inclusions are present in FTDP-17 brains. The pattern of inheritance in FTDP-17 is autosomal dominant with an early age of onset (45-65 years). The identification of mutations in the tau gene in FTDP-17 demonstrated that there is a direct link between tau dysfunction and neurodegeneration. To date, 14 missense mutations, a three base pair deletion (deltaK280) and seven splice site mutations have been found in over 50 FTDP-17 families from different ethnic groups. All of the known mutations occur in the C-terminal end of tau, with the majority affecting exons 9 12, which encode the microtubule-binding repeats. The mutations have multiple effects on the biology and function of tau. These varied pathogenic mechanisms most likely explain the wide range of clinical and neuropathological features observed in different families with FTDP-17. PMID- 12572917 TI - The heterogeneity of neuropathological changes in amyotrophic lateral sclerosis. A review of own autopsy material. AB - As an approach to the validation of different pathological findings in ALS, we have reviewed the histopathological slides of 24 cases (12 men, 12 women) of autopsy-confirmed ALS which had been treated in our centre (18 definite, 3 probable and 3 possible ALS). The mean age and survival was 57.1 years and 23.5 months respectively. The slides (from motor cortex, basal ganglia, brain stem and spinal cord) were reviewed with special attention paid to the severity of motor cortex and corticospinal tract degeneration, infiltrations of macrophages, atrophy of lower motor neurones, accumulation of lipofuscin, chromatolysis and gliosis. We also searched for the occurrence of axonal spheroids and various kinds of inclusions, like Bunina bodies, large hyaline, Lewy-body-like inclusions and even larger "hyaline balls", and ubiquitinated inclusions. The histological examination revealed considerable differences between particular cases. Different kinds of inclusions were found in 19 out of 24 cases. A shorter duration of the disease was found in the cases with numerous macrophages. The apparent histological degeneration of the motor cortex was found in cases with longer duration of the disease. However, the differences were not significant and no correlation between the occurrence of any type of inclusion or any other pathological finding and the duration of the disease was found. Whether apparent pathomorphological "heterogeneity" reflects some true differences in the pathogenesis of ALS is disputable. PMID- 12572918 TI - Morphological analysis of active microglia--rod and ramified microglia in human brains affected by some neurological diseases (SSPE, Alzheimer's disease and Wilson's disease). AB - The activation of microglial cells in pathological conditions is manifested primarily by their proliferation, as well as by the occurrence of a new morphological form--rod microglia. In the present study immunohistochemical identification of rod microglial phenotype against ramified microglia was performed on segments of 17 brains derived from 7 cases of encephalitis of viral aetiology (including 5 SSPE cases), 6 cases of Wilson's disease and 4 cases of Alzheimer's disease. Segments from frontal, temporal and occipital lobes, cerebellum and brainstem were subjected to histological, histochemical and immunohistochemical reactions. The presence of activated rod and ramified microglia was observed in sections derived from all structures of the brains under study. Both morphological forms of activated microglia reacted to antibodies: HLA II, CD68, HAM56 and lectin RCA-1. Expression of HLA II molecules was less intensive on the surface of microglial rod cells. A positive reaction to PCNA antibody was mainly observed in rod/elongated/cylinder-shaped nuclei, which is a characteristic feature of rod microglia. In the study material, the localisation of microglial processes seemed to depend rather on the structural topography of the cell in the brain than on the nuclear shape of the activated microglial cell. Our observations revealed a strong similarity between immunohistochemical phenotypes of both morphological forms of microglia with the indication that rod microglia is a first developmental form of activated microglia. PMID- 12572919 TI - The pattern of irreversible brain changes after cardiac arrest in humans. AB - Irreversible brain damage resulting in sustained post-arrest coma is a most serious complication of CA in resuscitated patients. In this study we present the neuropathological examination performed on 17 brains of patients who had remained unconscious for variable post-resuscitation periods and died after survival time ranging from 1 day to 36 days following cardiac arrest. Two main types of irreversible brain lesions were observed. The first one consisted of primary ischaemic selective disseminated neuronal death appearing in the form of acidophilic neurones with consecutive neuronal loss and reactive astroglial activation in the brain regions vulnerable to ischaemia or in more widespread distribution. The second included microinfarcts or confluent areas of pancellular necrosis associated with perivascular and diffuse tissue sponginess and was considered as secondary to post-resuscitation systemic and regional circulatory disturbances. These lesions showed multifocal, perivascular and laminar distribution with apparent predilection to the cortical border zones of arterial supply territories and were associated with disintegration and loss of GFAP reactive astrocytes. Continuity between the areas of selective neuronal loss and focal tissue pannecrosis with corresponding patchy disappearance of GFAP- and VIM positive astrocytes indicates the contribution of astroglia in the progression of postischaemic brain lesions. Both types of changes, evaluated in the neocortex and hippocampus, were more intensive and heterogeneous in the patients dying after prolonged coma duration than in those with recovery of consciousness within the first days after resuscitation. PMID- 12572920 TI - Is the normal content of sulfhydryl groups attributable to sparing from dystrophic pathology in dystrophin-deficient muscles? AB - Deficiency of dystrophin in skeletal muscles is supposed to be responsible for all the symptoms associated with Duchenne dystrophy (DMD) and Becker dystrophy (BMD). The dystrophin-deficient mdx mice, however, are clinically almost asymptomatic. Hence, other factor(s) might be responsible for the muscle pathology in DMD and BDM. As sulfhydryl groups are involved in maintaining the structure of membranes and the protein-phospholipid interactions, total, protein bound and free sulfhydryl groups (-SH) in DMD, BMD, limb-girdle dystrophy (LGMD) and the mdx mice muscles have been determined. A significant decrease of total and protein-bound -SH groups content and an increased proportion of free -SH groups in DMD and BMD was found. In LGMD the changes of total and protein-bound SH groups content were less expressed. In the mdx mice muscles the content of -SH groups was generally normal, only a higher than normal proportion of free-SH groups content in old and senile animals, especially in their diaphragm, was present. To test the sarcolemmal integrity, albumin/creatine kinase (CK) influx/efflux was determined. In early stages of DMD and BMD the albumin influx was increased. In advanced stages of these diseases albumin influx was not observed. In LGMD albumin penetration was present only in a few fibres. CK efflux in vitro was increased both in early and advanced DMD cases. In BMD and LGMD CK efflux was increased only in early stages of the diseases. In mdx mice an increased influx/efflux of albumin/CK was stated in adult animals. The changes persisted in the mdx hind limb muscles up to the senile age of the animals. In the mdx diaphragm of senile animals albumin did not penetrate the muscle cell and no increase of CK efflux was observed. It is suggested that changes in the distribution of -SH groups take part in the molecular disorganisation of the sarcolemma in course dystrophinopathies. Normal content of the sulfhydryl groups is supposed to be attributable to sparing from dystrophic pathology in dystrophin deficient mdx mice muscles. PMID- 12572921 TI - IL-15 is elevated in sera of patients with relapsing-remitting multiple sclerosis. AB - Interleukin-15 is a novel cytokine produced by monocytes/macrophages and sharing several biological activities with IL-2. IL-15 induces T cell proliferation, enhances natural killer (NK) cell cytotoxicity and also stimulates B cells to proliferate and secrete immunoglobulins. The purpose of our study was to measure IL-15 levels in the serum and CSF of 21 patients with relapsing-remitting form of MS, 9 with active gadolinium enhancing lesions in MRI, 12 without enhancing MRI lesions and to compare the results with the control group. IL-15 levels were measured by ELISA. We found a significant increase of IL-15 in the sera of patients with MS in comparison with the control group consisting of 8 patients with tension headache. IL-15 serum levels were highest in patients with active, gadolinium enhancing lesions. IL-15 CSF levels were low and there was no difference between studied groups. The results may suggest the contribution of IL 15 in the immunopathogenesis of multiple sclerosis. PMID- 12572922 TI - Apoptosis in non-astrocytic brain tumours in children. AB - Apoptosis is a process of great significance in cell biology. It plays a crucial role in both physiological and pathological conditions. An example is a tumour growth that is based on a subtle balance between cell division and cell death. Previously necrosis was considered to be the major type of cell death in tumours. Many recent investigations have focused on apoptosis, a phenomenon of great importance, and it is this type of cell death that is frequently "chosen" by a moribund cell. The objects of our interest were central nervous system (CNS) tumours, in which we estimated the number of apoptotic cells and sought for any correlation between the intensity of apoptosis and other markers of proliferation. Therefore, we studied CD34, the marker of angiogenesis, and Ki67, the marker of cell proliferation. We investigated 19 medulloblastomas and 15 ependymomas, among which 6 were anaplastic. We used in-situ labelling of DNA fragments to detect apoptosis in paraffin-embedded tissues. The mean value of the apoptotic ratio (AR) for all examined brain tumours was 0.012 with a standard deviation (SD) of 0.032, for medulloblastomas 0.021 (SD 0.04), for ependymomas 0.001 (SD 0.002) and for anaplastic ependymomas 0.004 (SD 0.003). PMID- 12572923 TI - A case of pseudo-aprosencephaly. AB - We present a case of rare, severe brain malformations. In a microcephalic premature newborn only a small structure found in the base of the cranium corresponded to the prosencephalic part of the brain. The basal ganglia were lacking and the cortico-subcortical area changed into empty vesicles with only small fragments of cortical stripe. This picture authorises us to speak on the probably complex pathomechanism of anomalies: genetically caused malformations including underdevelopment of telencephalic nuclei and additional occurrence of extrinsic lesions leading to pseudo-aprosencephaly formation. PMID- 12572924 TI - Outcome of renal transplantation in patients with pauci-immune small vessel vasculitis or anti-GBM disease. AB - AIM: Pauci-immune small vessel vasculitis (SVV) and anti-GBM disease are the most common causes of rapidly progressive glomerulonephritis (RPGN) and they frequently lead to end-stage renal disease. For renal replacement therapy, renal transplantation is the preferred treatment option. However, in patients with glomerular diseases, the outcome of renal transplantation can be adversely affected by recurrence of the original disease. The information in the medical literature on the outcome of renal transplantation in patients with RPGN is limited because most data are derived from case studies and from studies involving a small number of patients. METHODS: We studied the outcome of renal transplantation in patients with pauciimmune SVV or anti-GBM disease, transplanted in our center between 1968 and 2000. Patient and graft survival were compared with a matched control group from our hospital. We specifically looked for any evidence of recurrent disease. RESULTS: Included in the study were 43 patients (31 male, 12 female) with a mean age (+/- SD) of 48 +/- 15 years at transplantation. Patients were diagnosed as Wegener's granulomatosis (n = 8), microscopic polyangiitis (n = 7), renal limited vasculitis (n = 18) and anti-GBM disease (n = 10). The average follow-up was 62 +/- 57 months. No graft was lost due to recurrence of the underlying disease. One patient with Wegener's granulomatosis had a relapse with only extrarenal manifestations 5 months after transplantation. Patient and graft survival at 5 years after transplantation were 77% and 60%. Survival rates were not significantly different from a matched control group of renal transplant patients with other underlying diseases, 79% and 56%, respectively. Patients with pauci-immune SVV or anti-GBM disease developed significantly more malignancies than the control group (p = 0.02). CONCLUSIONS: Recurrence of pauci-immune SVV and anti-GBM disease after transplantation is rare. Renal transplantation can be successfully performed in patients with pauciimmune vasculitis or anti-GBM disease. Physicians should be aware of the greater risk of developing malignancies, especially skin cancer. PMID- 12572925 TI - No association of the TGF-beta1 gene polymorphisms with the renal progression in autosomal dominant polycystic kidney disease (ADPKD) patients. AB - BACKGROUND: Two genetic loci, PKD I and PKD2, have been identified as being responsible for ADPKD, and PKD1 is known to be associated with a poor prognosis. However, the presence of an intrafamilial study clinical diversity suggests that there are disease-modifying loci. Because the mechanism ofthe renal failure in ADPKD includes a cystic growth and tubulointerstitial atrophy and fibrosis, we studied the associations between 2 polymorphisms in the TGF-beta1 gene, which are known to be associated with chronic tubulointerstitial inflammation, and ADPKD progression in Korean patients. PATIENTS AND METHODS: One hundred and twenty-five individuals who had ADPKD and 47 normal control subjects were genotyped by PCR RFLP, the T869C (Leu10Pro) variant of TGF-beta gene leader sequence was discriminated with MspA1I and the G915C (Arg25Pro) variants with Bg1I. Statistical significances were determined using the Chi-square test. RESULTS: The distribution of the alleles for the TGF beta1 Leu10Pro polymorphism in ADPKD was: T 54%, C 46%, which was similar to the Korean (56: 44, p = 0.887) and Western controls (65: 35). In addition, no differences were found between the ESRD and the non-ESRD groups (p = 0.888) or the early hypertension and the normotension groups (p = 0.249). The distribution of alleles for the TGF beta1 Arg25Pro polymorphism showed only the GG type which was different from the Western population controls (G:C = 90:10, p = 0.000). CONCLUSIONS: Our results suggest that the polymorphism at Arg25Pro of TGF-beta1 in the Korean population has an allele distribution different from that ofthe Western population and that the polymorphism at Leu10Pro of TGF-beta1 has no association with the renal progression in Korean ADPKD patients. PMID- 12572926 TI - Comparison between serum creatinine and creatinine clearance for the prediction of postoperative mortality in patients undergoing major vascular surgery. AB - BACKGROUND: Poor renal function prior to surgery is associated with increased risk for mortality in patients undergoing major vascular surgery. Traditionally, this function is assessed by serum creatinine concentration (SeCreat). However, SeCreat is also influenced by age, gender and body weight. Hence, creatinine clearance (C(Cr)) is considered to be a better reflection of renal function. This study was undertaken to explore the prognostic value of preoperative calculated Cc, compared to SeCreat for the prediction of postoperative mortality. PATIENTS AND METHODS: The study group comprised 852 consecutive patients who underwent elective major vascular surgery at the Erasmus Medical Center, Rotterdam. Preoperative C(Cr) was calculated based on the Cockroft-Gault equation using preoperative SeCreat, age, body weight and gender. Univariable logistic regression analyses were used to study the relation between preoperative SeCreat, C(Cr) and postoperative mortality. Furthermore, multivariable logistic regression analysis was applied to evaluate the additional predictive value of age, body weight and gender additional to SeCreat. The receiver operating characteristic (ROC) curve was determined to evaluate the predictive power of several regression models for perioperative mortality. RESULTS: Postoperative mortality was 5.9% (50/852) within 30 days of surgery. In a univariable analysis, 10 micromol/l increment of SeCreat were associated with a 20% increased risk of postoperative mortality (OR = 1.2, 95% CI, 1.1-1.3) with an area under the ROC curve of 0.64 (95% CI, 0.56-0.71). If age, gender and body weight were added, the area under the ROC curve increased to 0.70 (95% CI, 0.63-0.77; p < 0.001), indicating that these risk factors had additional prognostic value. Indeed, in a separate regression analysis 10 ml/min decrease in C(Cr) was associated with a 40% increased risk of postoperative mortality (OR = 1.4,95% CI, 1.2-1.5; ROC area: 0.70, 95% CI, 0.63-0.76). ROC curve analysis showed that the cut-off value of 64 ml/min for C(Cr) yielded the highest sensitivity/specificity to predict postoperative mortality. CONCLUSION: Preoperative SeCreat was strongly associated with postoperative mortality, and adding age, gender, and body weight to the model showed improved predictive power indicating that preoperative C(Cr) calculated with these data has additional prognostic value. PMID- 12572927 TI - Nutritional effects of carnitine supplementation in hemodialysis patients. AB - AIMS: Carnitine is involved in fatty acid metabolism and it is cleared by dialysis. As it plays a role in energy utilization and because malnutrition is a frequent complication of HD treatment, we studied the effects of carnitine supplementation on several nutritional parameters in HD patients. MATERIAL AND METHODS: The main selection criterion was a body mass index (BMI; body weight/(height)2) < 22 kg/m2. Fifty-three patients were enrolled to participate in this open and randomized study. For 6 months, 28 patients received 15 mg/kg of intravenous L-carnitine at the end of each hemodialysis (HD) treatment (Group A), the remaining 25 patients were controls (Group B). The measured parameters were the post-dialysis body weight, serum albumin concentration (nephelemetry), food intake assessed by a 3-day food questionnaire, nPNA (normalized protein equivalent of nitrogen appearance), creatinine generation, and anthropometry. RESULTS: Forty-five patients completed the study (Group A: 14 F/9 M, 66.7 years old; Group B: 11 F/11 M, 65.2 years old). At the beginning of the study, there were no differences between the groups for age, gender, HD duration, BMI, diabetes prevalence, plasma carnitine levels and measured nutritional parameters. 65.2% and 77.3% in each group were carnitine-deficient (plasma total carnitine level < 35 micromol/l). After 6 months of L-carnitine supplementation, none of the nutritional parameters had changed in either group, except that serum albumin concentration decreased in both groups. Dividing each group according to their respective median serum albumin concentrations, daily energy and protein intakes, creatinine generation or triceps skinfold thickness did not show any difference in the various nutritional parameters with or without carnitine supplementation. CONCLUSION: Carnitine supplementation, despite normalization of plasma carnitine levels, has no effect on the nutritional status of HD patients. PMID- 12572928 TI - Relationship between plasma level of parathyroid hormone and carboxymethyllysine in hemodialyzed patients--does it exist? AB - AIMS: Both parathyroid hormone and advanced glycated end products (AGEs) are uremic toxins. The present study aimed to examine the likely interrelationship between these compounds. METHODS: Seventy-four hemodialyzed patients (41 female, 33 male; mean age 47 +/- 2 years, mean duration on hemodialysis 36 +/- 6 months) were enrolled in this study. In all subjects, the body mass index (BMI) was calculated and total lean mass (TLM) and total fat mass (TFM) were assessed by dual X-ray absorptiometry. Blood samples for estimation of plasma calcium, phosphorus, carboxymethyl lysine (as marker of AGEs) and PTH-1-84 were obtained after overnight fasting, before subsequent hemodialysis session. RESULTS: BMI, TFM and TLM were 23.6 +/- 0.5 kg/m2, 16.3 +/- 1.0 kg and 46.3 +/- 1.1 kg, respectively. PTH plasma level (223 +/- 32 pg/ml) and plasma CML (1,837 +/- 84 ng/ml) were markedly elevated as compared with reference values. A significant positive correlation was found between TLM and CML levels (tau = 0.225; p = 0.04) and between plasma PTH and CML levels (tau = 0.224; p = 0.04). CONCLUSION: It seems likely that PTH and AGEs are interrelated. The pathophysiological relevance of this finding in the pathogenesis of uremic toxicity remains to be elucidated. PMID- 12572929 TI - Differential use of cardiac troponin T versus I in hemodialysis patients. AB - BACKGROUND: Cardiac troponin T (cTnT) is frequently elevated in asymptomatic hemodialysis (HD) patients and predicts increased cardiovascular morbidity and mortality. Compared to cTnT, cardiac troponin I (cTnI) has a shorter half-life. How this influences its diagnostic reliability in chronic HD patients is only partially known. PATIENTS AND METHODS: First, in a cross-sectional study cardiac troponins were measured in 31 asymptomatic HD patients. A third-generation cTnT assay was used. The rate of false positive tests and the intraindividual variability were determined. Second, in a retrospective analysis over 12 months all acute events with clinical suspicion for acute coronary syndrome (ACS) were analysed in the same patients to determine the diagnostic power of cTnT by receiver-operating curve (ROC) plot. RESULTS: Cross-sectional study: 9 of 52 (17%) cTnT and 0/52 cTnI (0%) tests were positive in asymptomatic HD patients with a low intraindividual variability. Retrospective analysis: 16 acute clinical events with determination of cTnT were recorded, and in 4/16 an ACS was diagnosed. Using a cut-off level of 0.1 microg/l, the cTnT test reached a sensitivity of 100%, a specificity of 42%, a positive predicitive value of 36% and a negative predictive value of 100%, using a cut-off level of 0.2 microg/l the corresponding values were 75%, 58%, 38% and 88%. CONCLUSIONS: Cardiac TnT, but only rarely cTnI, is elevated in a significant number of asymptomatic HD patients. For diagnosis of ACS in HD patients, a combination of cTnT and cTnI may be used, since the former has higher sensitivity and the latter higher specificity. A higher threshold value for cTnT in HD patients could further increase its diagnostic accuracy. PMID- 12572930 TI - Large atrial thrombus formation associated with tunneled cuffed hemodialysis catheters. AB - AIMS: In the existing literature, there is a paucity of data regarding large atrial thrombus (AT) formation occurring as a complication of tunneled cuffed hemodialysis catheter (TCC) use. This study was performed to determine the risk factors, mortality and the appropriate management of TCC-AT. METHODS: We report 6 new cases of TCC-AT and have amalgamated these data with data from 16 previously published cases of TCC-AT found by performing a PubMed literature search (total of 22 cases). Demographic data were collected prospectively over 2 years in 85 consecutive patients initiating hemodialysis who were using a TCC as their primary vascular access, so that comparisons could be made between the 6 patients with TCC-AT versus all patients with a TCC at our center. RESULTS: In patients with TCC-AT, the mean time from TCC insertion was 4.5 months, and infection was present at the time of diagnosis in 68% of cases. The mean thrombus size was 3.7 cm, range 1.5-8 cm. All but 1 case were visualized by echocardiography; the remaining case required magnetic resonance imaging. Management included TCC removal and thrombectomy (n = 9), TCC removal and anticoagulation (AC) (n = 6), TCC removal alone (n = 5), and no intervention (n = 2). The overall mortality was 27%, and 5 of the 6 deaths (83%) occurred in patients with bacteremia. The mortality associated with each management strategy was as follows: TCC removal and thrombectomy (0%), TCC removal and AC (33%), TCC removal alone (40%), and no intervention (100%). CONCLUSIONS: AT is a serious complication of TCC use in hemodialysis patients and may be associated with a high mortality rate. TCC-AT may occur more commonly than previously reported and therefore warrants a high index of suspicion. PMID- 12572931 TI - A preliminary survey of bacterial contamination of the dialysate circuit in continuous veno-venous hemodialysis. AB - AIMS: The problem of dialysate bacterial contamination has not been defined in continuous renal replacement therapy. We assessed the bacterial integrity of source bicarbonate dialysate (study 1) and the continuous veno-venous HD (CVVHD) bicarbonate dialysate circuit (study 2). METHODS: Study 1: 50 ml dialysate were collected from 41 bags randomly selected from 150 consecutively made dialysate bags, immediately after manufacture or after 24, 48 or 72 h. Study 2: 10 ml dialysate were drawn from 4 sample points ranged along the dialysate circuit in 18 therapies (mean duration 119.5 +/- 72.0 h). All points were sampled at therapy start then daily, bar the proximal point which was sampled after each dialysate bag change. All dialysate samples underwent Gram stain and aerobic/anaerobic culture. Samples over 10 ml were cultured after centrifugation (15 min, 4,000 rpm). A disseminated contamination (DC) involved > or = 1 sample point at a time and/or was sustained over time. RESULTS: Study 1: One bag was culture-positive (staphylococcal/diphtheroid growths; 48-h sample). Study 2: Six DCs developed in 6 therapies (1 at therapy end, 5 sustained to therapy end (duration 57.25 +/- 45.95 h), 5 with Gram-negative bacilli, all involving reported growths of > or = 1,000 cfu). Dialyzer-inclusive dialysate circuit changes were more frequent in non-DC therapies (change rate: DC, 0.08 +/- 0.12/day, non-DC, 0.34 +/- 0.23, p = 0.02, permutation tests with general scores) but did not entirely prevent DC or alter it once underway. CONCLUSIONS: Sustained bacterial contamination of bicarbonate-based CVVHD is common and could relate to the completeness of dialysate circuit change. The importance of technique and regular quality control is highlighted. PMID- 12572932 TI - Hemodialysis versus continuous veno-venous hemodiafiltration in the management of severe valproate overdose. AB - Valproate intoxication is a relatively common clinical problem that can result in coma, respiratory depression, pancytopenia, hemodynamic instability and death [Fernandez et al. 1996, Franssen et al. 1999]. The drug's relatively low molecular weight, small volume of distribution and saturable protein-binding render it potentially amenable to extracorporeal removal (hemofiltration, hemodialysis or hemoperfusion), but published experience is scarce. This report describes a woman with a potentially fatal sodium valproate overdose, who did not respond to continuous veno-venous hemodiafiltration, but was successfully treated with low-flux hemodialysis. Based on our experience, we recommend hemodialysis for serious valproate intoxication. PMID- 12572934 TI - Acute poststreptococcal glomerulonephritis mimicking Henoch-Schonlein purpura. PMID- 12572933 TI - Cardiac and pulmonary calcification in a hemodialysis patient: partial regression 4 years after parathyroidectomy. AB - AIMS: The reversibility of extraskeletal calcifications in dialysis patients is an important and unresolved issue. Although periarticular calcifications have been shown to be reversible, little data are available on vascular or parenchymal calcifications. CASE HISTORY: A patient on maintenance hemodialysis with severe hyperparathyroidism, hypercalcemia and hyperphosphatemia was admitted to undergo parathyroidectomy. A preoperative total body bone scintigraphy was performed to better evaluate a lytic lesion in the pelvis, the histology of which proved to be a "brown tumor". The scan showed the typical findings of renal osteodystrophy, but also a diffuse extra-skeletal uptake of bone tracer in the lungs, kidneys, femoral arteries and myocardium. After surgery, good control of serum calcium, phosphate (Ca x P product < 50 mg2/dl2) and PTH levels was maintained during 4 years of follow-up. Bone scans were repeated after 2 and 4 years, showing marked improvement of periarticular uptake at the ends of long bones. Extraosseous calcium deposition was still markedly evident, but progressively decreased (at 4 years: heart -36%, lungs -18%). CONCLUSION: In this dialysis patient, extraskeletal calcification of visceral organs (particularly in the heart and the lungs) due to prolonged hypercalcemia and hyperphosphatemia was partially reversible by parathyroidectomy followed by good long-term control of serum phosphate and calcium. PMID- 12572935 TI - Long-term outcome of acute tubulointerstitial nephritis: report of a case. PMID- 12572936 TI - Hemolytic uremic syndrome and cattle. PMID- 12572937 TI - Trichosporon inkin peritonitis in a patient on continuous ambulatory peritoneal dialysis returning from the Caribbean. PMID- 12572938 TI - Systematic review of the distribution of the major vector-borne parasitic infections in dogs and cats in Europe. AB - On the basis of a systematic review of the literature, this paper provides the first evidence-based maps of the distribution of the major vector-borne parasitic infections of dogs and cats in Europe. From an initial survey of 894 publications, data from 268 were analysed, summarised and collated. Prevalence data were used to calculate the force of infection to provide distribution and incidence maps for canine infections with Leishmania infantum, Dirofilaria immitis, Dirofilaria repens, Babesia canis and Ehrlichia canis. There was little information from some areas but, in spite of its acknowledged incompleteness, the study provides an evidence-based framework upon which to assess the risks of infection, and will provide a basis, by correlation with climatic and vegetation data, to derive more comprehensive risk assessment maps for Europe. To clinicians in both endemic and non-endemic countries, it will be of assistance in decision making with respect to diagnosis and preventive measures. PMID- 12572939 TI - Treatment of canine parvoviral enteritis with interferon-omega in a placebo controlled field trial. AB - The clinical efficacy of a recombinant feline interferon (IFN) (type omega) was evaluated under field conditions for the treatment of dogs with parvoviral enteritis. In this multicentric, double-blind, placebo-controlled trial, 94 dogs from one to 28 months old were randomly assigned to two groups which were treated intravenously either with IFN (2.5 million units/kg) or placebo once a day for three consecutive days, and monitored for clinical signs and mortality for 10 days. Each dog received individual supportive treatment The data from 92 interpretable cases (43 IFN-treated and 49 placebo) showed that the clinical signs of the IFN-treated animals improved significantly in comparison with the control animals, and that there were only three deaths in the IFN group compared with 14 deaths in the placebo group (P = 0.0096) corresponding to a 4.4-fold reduction. Alternative analyses of the data taking into account the prior vaccination status of the dogs against canine parvovirus suggested that the IFN therapy resulted in a 6.4-fold reduction in mortality (P = 0.044) in the unvaccinated cohort, a significant reduction when compared with the vaccinated cohort. PMID- 12572940 TI - Four cases of equine polysaccharide storage myopathy in the United Kingdom. AB - This paper describes four cases of equine polysaccharide storage myopathy which were confirmed by histological examination of muscle biopsy specimens. The horses were of mixed breeding, with warmblood and thoroughbred dominating. They all had recurrent episodes of rhabdomyolysis, indicated by clinical signs and increased plasma levels of muscle enzymes. They were managed conservatively and have continued athletic careers despite their disease. PMID- 12572941 TI - Sarcoptic mange (Sarcoptes scabiei var vulpes) in a red fox (Vulpes vulpes) population in north-west Surrey. PMID- 12572942 TI - Influence of quinidine and flecainide on autonomic nervous activity in thoroughbred horses. PMID- 12572943 TI - Disposal of veterinary sharps. PMID- 12572944 TI - Transmission of bluetongue virus in Italy. PMID- 12572945 TI - Mastitis and somatic cell counts. PMID- 12572946 TI - Veterinary forensic evidence. PMID- 12572947 TI - Hip dysplasia in dogs. PMID- 12572948 TI - Cutaneous infection in a cat caused by Mycobacterium avium complex serovar 6. PMID- 12572949 TI - RCVS library and information service membership scheme. PMID- 12572950 TI - Vaginal lactobacilli: self- and co-aggregating ability. AB - Lactic acid bacteria are the dominant bacteria of the vaginal tract in healthy women. Lactobacillus species form a barrier population that protects from pathogen colonisation by mechanisms that include adhesion to epithelial surfaces, self-aggregation and co-aggregation. In this study, factors involved in the self aggregating ability of vaginal lactobacilli and in the co-aggregation of these microorganisms with Candida spp. are studied. Both self-aggregation and co aggregation are monitored quantitatively by the decrease in the absorbance of suspensions of the microorganisms and qualitatively by light microscopy. The self aggregating ability of four vaginal lactobacilli was shown to be caused by a peptide or protein sensitive to trypsin. However, in self-aggregating Lactobacillus acidophilus CRL 1294 the factor was resistant to trypsin and sensitive to pepsin. Among self-aggregating lactobacilli, L. acidophilus CRL 1294 and L. salivarius CRL 1328 were able to co-aggregate with Candida spp. The co aggregating factor for both strains proved to be peptide of the surface and a peptide on the bacterial surface, while the receptor on the yeast was a carbohydrate. Co-aggregation of both lactobacilli and Candida spp. was inhibited by the addition of mannose but was not affected by other carbohydrates. Self and co-aggregation factors were not able to induce aggregation in non-aggregating lactobacilli. PMID- 12572951 TI - Molecular and immunological characterisation of Fasciola species. AB - Fasciola hepatica and F. gigantica are polymorphic liver flukes that show considerable overlap between species, and various protein separation techniques have been used as alternative means of differentiation. Acid and alkaline polyacrylamide gel electrophoresis (PAGE) show differences between F. hepatica and F. gigantica. Following SDS-PAGE, F. hepatica proteins are characterised by the presence of eight major peptide bands, with molecular weights estimated as 48, 45, 43.5, 37, 33, 29, 27 and 25.5 kDa. In contrast, F. gigantica shows only five major protein bands of 57.6, 54, 48, 29 and 27 kDa. Isoelectric focusing (IEF) demonstrates 17 bands from F. hepatica and 22 bands from F. gigantica between pH 3.5 and pH 10. Although many bands appear common to both species, some are species-specific. Six cases of human acute fascioliasis diagnosed clinically, haematologically and immunologically are also studied. Gel immunodiffusion and immunoelectrophoresis, using adult F. hepatica and F. gigantica antigens, are used to determine the species, and indicate that the antisera are more specific for F. hepatica. PMID- 12572953 TI - How much tissue is present in archival paraffin blocks and slides? AB - In order to inform the debate about tissue blocks and slides introduced by the Retained Organs Commission, a study is undertaken to determine the percentage weight of tissue present in the surgical archive in the cellular pathology department of a district general hospital. When original, unprocessed tissue weight is expressed as a percentage, based on the weight of 100 archival paraffin blocks, values range from 0.2% to 41.5%. When the corresponding archival slides are also included, the values fall as low as 0.1% (i.e. up to 99.9% of the stored archival material for a piece of processed tissue could be non-human material). The results are used to make a case for including archival histological material as part of the patient's clinical record, although it is accepted that this study was performed only on surgical tissue. PMID- 12572952 TI - Evaluation of melanocytic neoplasms: application of a pan-melanoma antibody cocktail. AB - Incidence of malignant melanoma (MM) is rising rapidly throughout the Western world, and the number of melanocytic lesions removed for histological assessment has increased. MM can present with a myriad of histological appearances that make diagnosis problematic, particularly when dealing with metastatic deposits. Immunohistochemical diagnosis relies on a panel of antibodies comprising polyclonal S100 protein and the monoclonal antibodies HMB 45, MART-1, tyrosinase and, to a lesser extent, NKIC3. Confirmation of problematic cases relies on the use of polyclonal S100 protein, as its sensitivity has yet to be matched by any monoclonal antibody. The introduction of a potentially valuable pan-melanoma cocktail, composed of HMB 45, MART-1 and tyrosinase, is examined in 50 primary cutaneous malignant melanomas, five desmoplastic malignant melanomas (DMM), 35 benign naevi, 20 metastatic malignant melanomas, 10 basal cell carcinomas (BCC) and 10 squamous cell carcinomas (SCC) and compared to individual immunolabelling with S100 protein, HMB 45, MART-1 and tyrosinase. All BCCs and SCCs were negative with all antibodies. S100 protein, MART-1, tyrosinase and the pan-melanoma cocktail were positive for all cases of benign naevi. HMB 45 labelled all junctional and compound naevi, five of the eight intradermal naevi and five of the seven blue naevi. All 50 primary cutaneous MMs were positive with S100 protein, 49/50 with the pan-melanoma cocktail and tyrosinase, 47/50 with MART-1 and 46/50 with HMB 45. Of the five cases of DMM, all were positive with S100 protein and three of the five were positive with HMB 45, MART-1, tyrosinase and the pan-melanoma cocktail. In the case of metastatic MM, all 20 cases were positive with S100 protein, the pan-melanoma cocktail and tyrosinase. MART-1 was positive in 19/20 cases and HMB 45 in 17/20 cases. The pan-melanoma cocktail showed a high sensitivity for all forms of MM and should be considered a complementary marker to polyclonal S100 protein. Results confirmed that currently there is no alternative antibody available to match the sensitivity of polyclonal S100 protein for immunolabelling DMM. PMID- 12572954 TI - Effects of storage on the physicochemical and antibacterial properties of human milk. AB - Storage of human milk for limited periods of time is unavoidable in neonatal units and also in the home where increasing numbers of mothers go back to work soon after delivery. Many mothers, convinced of the importance of prolonged exclusive or complementary breast-feeding, often express and store human milk for use during the period of separation. This study examines the effects of different storage methods on the pH and some antibacterial activities of human milk. Portions of milk and colostrum samples from healthy lactating mothers were stored at 4 degrees C and -20 degrees C for periods ranging from one day to 12 weeks. The stored milk samples were analysed for pH, bactericidal and bacteria sequestration activities against a serum-sensitive Escherichia coli, and compared with freshly collected samples, with and without EDTA. Milk became progressively more acidic during storage. While the bactericidal activities of refrigerated samples diminished rapidly, up to two-thirds of the original activity level was maintained by freezing for up to three months. The ability of milk fat globule membrane to adhere to suspended bacteria was gradually lost in frozen milk samples, while it was greatly enhanced during the first few days in refrigerated samples, before declining sharply. This study shows that loss of bactericidal activity in refrigerated milk is well compensated for by enhanced bacteria sequestration activity, and allays any fears that might arise concerning the suitability of stored human milk for infant consumption. PMID- 12572955 TI - Synthetic organic hard capsule colouring agents: in vitro effect on human true and pseudo-cholinesterases. AB - Hard capsules are made of pure gelatin and small quantities of additives, including colouring agents permitted for use in food. In this study, the effects of three colouring agents (sunset yellow, quinoline yellow and erythrosine) on true and pseudo-cholinesterases (ChE) are assessed in erythrocytes and plasma, respectively. Results indicated that the synthetic compounds affected both true and pseudo ChE activity. The concentration of sunset yellow which caused 50% inhibition (IC50) of true ChE was about 64% that of pseudo-ChE; for erythrosine, IC50 was approximately the same for both true and pseudo-ChE; and for quinoline yellow, IC50 for true ChE was 25% of pseudo-ChE, although its effect on both true and pseudo-ChE was greater than seen with the other two dyes. Inhibitions of both true and pseudo-ChE were of mixed type (competitive and non-competitive). The enzyme-inhibitor dissociation constant (Ki) indicated that quinoline yellow was most potent and erythrosine was least potent out of the three compounds. Inhibition of both true and pseudo-ChE by each of the three dyes was abolished by dialysis, indicating that the effects were reversible. PMID- 12572956 TI - Diagnosis of visceral leishmaniasis: comparative potential of amastigote antigen, recombinant antigen and PCR. AB - Development of simple, economical and non-invasive tests for the early diagnosis of visceral leishmaniasis (VL) or kala-azar (KA) remains a challenge, and serological studies based on antigen prepared from the amastigote stage of Leishmania donovani, the stage that causes infection, are lacking. In the present study, circulating antibodies to total antigen isolated from the promastigote and amastigote stages of the parasite, as well as to recombinant K39 (rK39) antigen, are measured by enzyme-linked immunosorbent assay (ELISA) and the results compared with a polymerase chain reaction (PCR) test for KA diagnosis. In 116 samples of KA examined, the amastigote antigen gave significantly higher mean absorbance values in ELISA than did the promastigote antigen. The sensitivity for KA detection was significantly higher using the amastigote antigen (94%) than the promastigote antigen (90.5%). Analysis in 91 controls showed that specificity was higher with amastigote antigen (92.3%) than with promastigote antigen (86.8 89.0%). Reliability of ELISA diagnosis with amastigote antigen was only marginally lower than that with rK39 ELISA or with the PCR test. Easy availability and low cost of indigenous amastigote antigen, together with the simplicity of ELISA compared with PCR, make ELISA based on amastigote antigen a promising choice for the diagnosis of KA. PMID- 12572957 TI - Electrical impedance measurement as an endpoint detection method for routine coagulation tests. AB - A whole-blood platelet aggregometer is adapted to measure electrical impedance changes in plasma during prothrombin time (PT) estimations. The impedance curve shows an acceleration phase, which is comparable to the absorbance curve acceleration phase associated with the onset of coagulation. The amplitude of the impedance change correlates with the fibrinogen concentration of the plasma. Statistical analysis of PT derived by absorbance and impedance changes shows a significant difference between the two methods but a good correlation. The method is reproducible but laborious and requires attention to technique. Further investigation of the method utilising a more sensitive instrument and redesigned electrodes is indicated. It may also be possible to modify reagent systems to optimise impedance changes. PMID- 12572958 TI - Non-microscopic method for malaria diagnosis using OptiMAL IT, a second generation dipstick for malaria pLDH antigen detection. AB - Rapid diagnostic tests for malaria are now a commonly used procedure for malaria diagnosis. New or improved devices need to be evaluated against a recognised gold standard procedure and subjected to conditions of temperature and humidity that may affect their performance. The OptiMAL 48 RDT has now been available commercially for several years and a second-generation OptiMAL IT test is now coming onto the market. In this study the problems associated with the routine use of OptiMAL 48 is investigated and its performance compared with a second generation individual test, OptiMAL IT. Sensitivity and specificity for detection of all malaria species for both tests were comparable but loss of sensitivity of the test strips due to humidity or temperature found with the routine use of OptiMAL 48 was not seen with the individual OptiMAL IT. False-positive results for Plasmodium falciparum, seen in two negative blood samples, were attributed to the presence of high levels of heterophile antibodies. PMID- 12572959 TI - Bioterrorism: an overview. AB - Bioterrorism has reached the forefront of the public imagination following recent events across the world. The disaster of 11 September 2001, followed by anthrax letters sent via the US postal system and now renewed tension over Iraq have all brought the possibility of bioterrorism closer. A number of biological agents could be used in a terrorist attack, including anthrax, plague, smallpox and botulinum toxin. The serious diseases that these agents produce have been brought under control in the developed world; however, a lack of protective immunity against such diseases could cause considerable morbidity and mortality if used in a terrorist attack. This essay provides a background to bioterrorism, discusses many of the current points of interest and gives an update to the economic consequences of such an attack. PMID- 12572960 TI - Role of apoptosis in the pathogenesis of B-cell chronic lymphocytic leukaemia. AB - B-cell chronic lymphocytic leukaemia (B-CLL) is a clinically heterogeneous disease characterised by the accumulation of a clonal population of B lymphocytes. This accumulation is considered to result from the prolonged survival of B-CLL cells arrested in the G0 stage of the cell cycle. However, when cultured in vitro, B-CLL cells die rapidly by apoptosis. It is now clear that a number of factors can delay or postpone the onset of apoptosis, including a number of cytokines and direct contact with different cell types. Although many drugs are now known to cause clinical improvement in B-CLL by causing apoptosis of B-CLL cells, in only a few cases have biological mechanisms been reported to have similar effects. It is now important to understand the role of these mechanisms in the pathogenesis and progression of B-CLL, and to devise strategies to exploit them for therapeutic use. PMID- 12572961 TI - Aggressive treatment in the terminally ill: right or wrong? PMID- 12572962 TI - The confusion about confusion. PMID- 12572964 TI - The Broward County jails hospice program: hospice in the jail. AB - Broward County's jail hospice program initiated service in 1995. Infirmary services for both men and women in the jail system are housed in one of Broward County's four jail sites. The County, in partnership with the local Medicare certified hospice, Hospice by the Sea, provides care to inmates both in the detention center and on release to the community. Compassionate release, "community control," and expedited case disposition are sometimes utilized. Jail stays are typically shorter than prison sentences. Consequently, expedient identification and enrollment of hospice-appropriate detainees differentiates hospice in a jail setting from prison hospice. Initial attitudinal resistance to hospice services for detainees has for the most part been overcome. Ongoing challenges largely consist of institutional impediments regarding supplies, special dietary needs of hospice patients, visitation procedures, and potential patient isolation. PMID- 12572963 TI - Palliative care in phase 1 trials: an ethical obligation or undue inducement? PMID- 12572965 TI - The Maryland Division of Correction hospice program. AB - The Maryland Division of Correction houses 24,000 inmates in 27 geographically disparate facilities. The inmate population increasingly includes a frail, elderly component, as well as many inmates with chronic or progressive diseases. The Division houses about 900 human immunodeficiency virus (HIV)-positive detainees, almost one quarter with an acquired immune deficiency syndrome (AIDS) diagnosis. A Ryan White Special Project of National Significance (SPNS) grant and the interest of a community hospice helped transform prison hospice from idea to reality. One site is operational and a second site is due to open in the future. Both facilities serve only male inmates, who comprise more than 95% of Maryland's incarcerated. "Medical parole" is still the preferred course for terminally ill inmates; a number have been sent to various local community inpatient hospices or released to the care of their families. There will always be some who cannot be medically paroled, for whom hospice is appropriate. Maryland's prison hospice program requires a prognosis of 6 months or less to live, a do-not-resuscitate (DNR) order and patient consent. At times, the latter two of these have been problematic. Maintaining the best balance between security requirements and hospice services to dying inmates takes continual communication, coordination and cooperation. Significant complications in some areas remain: visitation to dying inmates by family and fellow prisoners; meeting special dietary requirements; what role, if any, will be played by inmate volunteers. Hospice in Maryland's Division of Correction is a work in progress. PMID- 12572966 TI - Factors associated with lower rates of in-hospital death. AB - OBJECTIVES: Examine individual and statewide factors associated with hospital as location of death. DESIGN: An observational study using large national databases. PARTICIPANTS: All 1997 U.S. decedents (all ages, all causes.) RESULTS: The West has the lowest percentage of deaths occurring in an acute care hospital, and the highest percentage of deaths at home. Increased hospital bed availability is significantly associated with greater chance of dying in an acute care hospital. Hispanic, nonwhite, and less educated decedents are more likely to die in a hospital (as opposed to home or nursing home). CONCLUSIONS: Hospital bed availability profoundly influences the likelihood of dying in a hospital. Individual characteristics such as race and education affect location of death as well. These three factors account for approximately 75% of the trend in the West to a lower in-hospital death rate and more deaths at home. Additional factors may include formal out of hospital orders to limit transfer. PMID- 12572967 TI - Estimation of confusion prevalence in hospice patients. AB - BACKGROUND: Confusion is common among ill patients and has broad consequences for their care and well-being. The prevalence of confusion in hospice patients is unknown. OBJECTIVES: Describe the prevalence, severity, and manifestations of nurse-identified confusion and estimate the prevalence of delirium in hospice patients. DESIGN: Cross-sectional descriptive study. SETTING: Nineteen hospices in the Population-based Palliative Care Research Network (PoPCRN). PATIENTS: Adult patients receiving care from participating hospices, February 15 to April 1, 2000. MEASUREMENT/ANALYSIS: Hospice nurses estimated prevalence, severity, behavioral manifestations, and consequences of confusion during the preceding week. Confused and nonconfused patients were compared using standard bivariate and stratification techniques. Logistic regression identified manifestations associated with problematic confusion. RESULTS: Median age of the 299 patients was 78 years; 59% were female, 52% lived at home, and cancer was the most common diagnosis (54%). Fifty percent were confused during the preceding week, 36% of those were severely confused or disabled by confusion. Compared with nonconfused patients, confused patients were less likely to have cancer (64% vs. 43%, p < or = 0.001) and more likely to live in nursing home/assisted living (21% vs. 33%, p < or = 0.01). Disorientation to time or place, impaired short-term memory, drowsiness, and easy distractibility were common manifestations of confusion. When present, confusion caused a problem for the patient, someone else, or both 79% of the time. Inappropriate mood, cancer diagnosis, agitation, and age were the variables predicting problematic confusion. Only 14% of confused patients met criteria for delirium. CONCLUSIONS: Confusion among hospice patients was common, frequently severe, and usually problematic. PMID- 12572968 TI - Objectives for advance care planning. AB - Identifying objectives for advance care planning (ACP) is an important step toward improving care at the end of life. Previous studies of ACP have used many different measures of success. However, there has been no consensus on what should be the objectives for ACP. Lack of attention to specific objectives for ACP may lead to ineffective communication and research. The first step to improving outcomes in ACP is to acknowledge the diversity of objectives that ACP may achieve. Health care providers, patients, and surrogates should identify and agree on common objectives for particular conversations. Various methods, conversations, and forms may be used to achieve these objectives over time. Clarifying objectives from various perspectives is an important step toward achieving the level of understanding necessary to make these difficult decisions. It is time for physicians to reconsider the way in which they think about and discuss ACP with patients. If we are to improve care at the end of life, future patient care, research, and education about ACP should proceed with specific objectives in mind. PMID- 12572969 TI - Age and gender differences in health care utilization and spending for medicare beneficiaries in their last years of life. AB - Men's and women's health care experiences differ as they age. While increasing attention has been focused on gender differences in health status, prevalence of illnesses, and access to quality care among older adults, little is known about differences in their health care in the last years of their lives. This paper uses claims data for a 0.1% random sample of Medicare beneficiaries who died between January 1, 1994 and December 31, 1998 to assess age and gender differences among Medicare-eligible adults in their utilization of health care services in the last year of life. Overall, age is much more important than gender in explaining most of the variation in end-of-life care. The combination of being a Medicare beneficiary and being sick enough to die appears to attenuate gender disparities in health care services utilization. PMID- 12572970 TI - Hospice experience and perceptions in nursing homes. AB - The purpose of this study was to describe the experiences and perceptions of hospice nurses caring for residents in long-term care facilities. The study used a fax-back survey to gather data from 69 hospice nursing and nurse managers in 24 hospices across one Midwestern state. Respondents reported negative experiences with pain management and care coordination in the nursing home setting. Although hospice is thought to be a benefit to residents, hospice staff report frustration in caring for nursing home patients, especially in trying to control pain. The study identifies opportunities for improvement in hospice and nursing home staff relationships. PMID- 12572971 TI - Experience with hospice: reflections from third-year medical students. AB - Nationally, there is a growing emphasis on experiential education and an interest in palliative care for health professionals. Hospice visits were added to the family medicine community rotation for third-year medical students to provide them with first-hand exposure to the hospice experience. Seven significant themes emerged from the students' reflection papers: (1) the value of hospice and the supportive role of the hospice team, (2) the value of empathy, (3) the distinction between acceptance and resignation, (4) the changing face of hope, (5) an understanding of death as a natural event, (6) the quality of family caregiving, and (7) the role of the physician in caring for the dying. The implications for medical education discussed include the value of home visits as a setting for experiences in palliative care and the effectiveness of narrative assignments for encouraging student reflection. PMID- 12572972 TI - End-of-life care for terminally ill participants in clinical research. AB - Efforts to improve end-of-life care in the United States have paid little attention to the unique concerns of participants in clinical research who are terminally ill. In this paper we focus attention on and offer an analysis of how to meet the needs of these individuals. To address their concerns, we consider how to reconcile two important tasks: providing optimal end-of-life care and conducting clinical research. First, we examine the inherent tension between the goals of medicine and the goals of science. Second, we focus more specifically on the tensions between a good death and conducting clinical research in patients with a short life expectancy. We examine six domains that have been suggested for measuring a good death: physical symptoms; psychological and cognitive symptoms; economic and caregiving needs; social relationships; spiritual beliefs; hopes and expectations. For each of these domains we examine how the goals of clinical research may conflict or coincide with taking care of a patient with a terminal illness. Finally, we offer suggestions to address these tensions: (1) modify the informed consent discussion for terminally ill participants in research; (2) build a palliative care component into clinical trials; (3) attend to the needs of family caregivers of terminally ill research subjects; (4) arrange for continuity of care so that dropping out of a trial does not jeopardize medical care; (5) train clinical investigators in end-of-life care; (6) develop a counseling strategy for terminally ill participants in clinical research. PMID- 12572973 TI - Fast facts and concepts #45. Medical management of bowel obstruction. PMID- 12572974 TI - Fast facts and concepts #47. What do I tell the children? PMID- 12572975 TI - Hole in one. PMID- 12572976 TI - In memoriam 2001. PMID- 12572977 TI - Integrating an end-of-life curriculum into the internal medicine clerkship. AB - OBJECTIVE: To integrate an end-of-life curriculum into the third-year internal medicine clerkship via communication skills training with standardized patients and a community hospice experience. DESCRIPTION: Clerkship directors identified a lack of exposure to end-of-life issues and hospice care as gaps in our third-year medical student curriculum. This curriculum was implemented to address these deficiencies. PMID- 12572978 TI - The Michigan Advanced Practice Nursing Palliative Care Project. AB - The emerging field of palliative care in the United States must respond to the health care needs of persons living and dying from advanced illness. The literature suggests that professional nursing education in palliative care is limited, inconsistent, and sometimes neglected. In response to these identified needs, the Michigan (MI) Advanced Practice Nurse (APN) Palliative Care Project was designed to integrate palliative education into existing graduate nursing curricula and to provide continuing education for postgraduate nurses. PMID- 12572979 TI - Volunteer patient advocacy: an interdisciplinary course on attending to patients at the end of life. AB - Little is known about the impact on health care students of early end-of-life care (EOLC) education and patient contact. We developed an EOLC course that allowed students to serve as volunteer advocates for patients at the end of life (EOL). We evaluated the course's acceptability to students and its effect on students' attitudes and beliefs. PMID- 12572980 TI - The Web-based worksheet: an opportunity for prompt, consistent, and expert feedback in a community-based hospice experience. AB - We faced a challenge in providing a consistent high-quality learning experience in hospice care, especially because our community-based medical school has students rotating in hospices in six separated communities and the number of faculty with expertise in palliative care is limited. To address these concerns, a Web-based worksheet with interaction with a central campus faculty member was designed for use in a hospice module in a family practice clerkship. PMID- 12572981 TI - Performance-based palliative care in the nursing home: closing the gap between "knowing" and "practicing". AB - This study evaluated the impact of an experiential educational model--combining traditional didactic techniques with applied performance-based methods--on changing nursing home staff attitudes and knowledge about palliative care, and effectuating positive changes in practice outcomes such that the delivery of palliative care is moved "upstream" in the disease/treatment trajectory. Two nursing homes participated. PMID- 12572982 TI - What role should the nephrologist play in the provision of palliative care? PMID- 12572983 TI - Palliative care in African American communities. PMID- 12572984 TI - The Harlem Palliative Care Network. PMID- 12572985 TI - Death and dying in the black experience. PMID- 12572986 TI - Animal models of hereditary iron transport disorders. PMID- 12572987 TI - Mechanism of iron toxicity. PMID- 12572988 TI - Role of non-transferrin-bound iron in the pathogenesis of iron overload and toxicity. PMID- 12572989 TI - Intracellular and extracellular labile iron pools. PMID- 12572990 TI - Cardioprotective effect of iron chelators. PMID- 12572991 TI - Results of long term iron chelation treatment with deferoxamine. PMID- 12572992 TI - Long term deferiprone chelation therapy. PMID- 12572993 TI - Iron chelator chemistry. PMID- 12572994 TI - Structure-activity relationships among desazadesferrithiocin analogues. AB - Desferrithiocin, a natural product iron chelator (siderophore), offers an excellent platform from which to construct orally active iron chelators which have a good therapeutic window. A systematic structure-activity study on desferrithiocin identified the structural fragments necessary for the compound's oral iron-clearing activity. There are strict requirements regarding the distance between the ligating centers; they cannot be altered without loss of efficacy. The thiazoline ring must remain intact. Benz-fusions, which were designed to improve the ligands' tissue residence time and possibly iron-clearing efficiency, are ineffective. The maintenance of an (S)-configured C-4 carbon is optimal in the design of desferrithiocin-based iron chelators. With this information in hand, alteration of the redox potential of the aromatic ring was initiated. Introduction of a hydroxy in the 4'-position of at least three different desazadesferrithiocin analogues resulted in moderate to small changes in iron clearing efficacy yet dramatic reductions in the toxicity of the compounds were observed. Although the toxicity studies of these desferrithiocin analogues are continuing, it is clear that it is possible to alter a siderophore in such a way as to ameliorate its toxicity profile while maintaining its iron-clearing properties. PMID- 12572995 TI - ICL670A: preclinical profile. PMID- 12572996 TI - Pyridoxal isonicotinoyl hydrazone and its analogues. PMID- 12572997 TI - Therapeutic potential of iron chelators in cancer therapy. AB - The success of DFO at markedly inhibiting the growth of aggressive tumors such as neuroblastoma and leukemia justifies interest in the development of chelators as anti-neoplastic agents. This is emphasized by the fact that DFO has suboptimal properties, namely poor membrane permeability and a very short serum half-life. More recently, the thiosemicarbazone chelator, Triapine, has entered a phase I clinical trial again confirming the potential of these compounds. Further studies examining the effects of chelators on neoplastic cells will not only be valuable in terms of identifing novel anti-cancer agents, but will also provide new information on the role of Fe in cell cycle control. PMID- 12572998 TI - Antimalarial effect of iron chelators. PMID- 12572999 TI - Fluid mechanics, cell distribution, and environment in CellCube bioreactors. AB - Cultivation of MRC-5 cells and attenuated hepatitis A virus (HAV) for the production of VAQTA, an inactivated HAV vaccine (1), is performed in the CellCube reactor, a laminar flow fixed-bed bioreactor with an unusual diamond-shaped, diverging-converging flow geometry. These disposable bioreactors have found some popularity for the production of cells and gene therapy vectors at intermediate scales of operation (2, 3). Early testing of the CellCube revealed that the fluid mechanical environment played a significant role in nonuniform cell distribution patterns generated during the cell growth phase. Specifically, the reactor geometry and manufacturing artifacts, in combination with certain inoculum practices and circulation flow rates, can create cell growth behavior that is not simply explained. Via experimentation and computational fluid dynamics simulations we can account for practically all of the observed cell growth behavior, which appears to be due to a complex mixture of flow distribution, particle deposition under gravity, fluid shear, and possibly nutritional microenvironment. PMID- 12573001 TI - Heat inactivation of mammalian cell cultures for biowaste kill system design. AB - A biowaste kill system was implemented to treat biological waste generated from a clinical manufacturing and R&D antibody facility. To confirm that design parameters of this continuous decontamination system are sufficient to inactivate mammalian cell culture waste, bench-scale experiments were conducted. The biowaste kill system heat inactivates mammalian cell cultures before they are piped to a neutralization tank and subsequently released to the sewage system. Heat inactivation of cells is accomplished by exposing cells to 80 degrees C for 1 min. Small-scale heat inactivation studies were performed on CHO, 293-HEK, and hybridoma cells. Cells at 1 x 10(6) cells/mL or 1 x 10(7) cells/mL were exposed to 37, 60, 70, or 80 degrees C for 0, 30, 60, and 120 s. Viability based on trypan blue exclusion method and ability to proliferate was assessed after exposure to heat. Data suggest that exposure of cells to 80 degrees C for 60 s is sufficient to inactivate these cultures before they are released to the sewage system. PMID- 12573000 TI - Sustained high-yield production of recombinant proteins in transiently transfected COS-7 cells grown on trimethylamine-coated (hillex) microcarrier beads. AB - The present study shows that COS-7 cells transiently transfected and maintained on positively charged (trimethylamine-coated) microcarrier beads synthesize recombinant protein at higher levels and for longer periods of time than cells transfected and maintained on polystyrene flasks in monolayer culture. Sustained, high-level synthesis was observed with secreted chimeric proteins (murine E selectin- and P-selectin-human IgM chimeras) and a secreted hematopoietic growth factor (granulocyte-macrophage colony-stimulating factor). Studies with green fluorescent protein indicated that the transfected cells attached more firmly to the trimethylamine-coated microcarriers than to polystyrene flasks. After 10-14 days in culture, most of the transfected cells detached from the surface of the polystyrene flasks, whereas most transfected cells remained attached to the microcarriers. The transiently transfected microcarrier cultures produced higher levels of protein per transfected cell due to this prolonged attachment. The prolonged attachment and higher output of transfected cells on microcarriers resulted in a 5-fold increase in protein production from a single transfection over two weeks. Thus, microcarrier-based transient transfection yields quantities of recombinant proteins with a significant savings of time and reagents over monolayer culture. PMID- 12573002 TI - Influence of carboxymethyl dextran and ferric citrate on the adhesion of CHO cells on microcarriers. AB - Due to the inherent risks of animal-derived raw materials, the biopharmaceutical industry has an increasing demand for serum-free and protein-free media for industrial cell culture bioprocesses. The absence of serum often changes the characteristics of mammalian cells, especially growth, productivity, and adherence properties. This study is mainly focused on the influence of media additives on cell adherence characteristics. An array of different carboxymethyl dextrans and different ferric citrate concentrations was tested with a number of CHO clones, using standard cell culture Roux-flasks and Cytoline 1 macroporous microcarriers. A prototype mixing system with controlled shear force input was developed as a screening system for adherence characteristics. The results of this evaluation revealed a negatively correlated dose-dependent influence on adhesion for ferric citrate. It was also found that certain carboxymethyl dextrans are capable of increasing the adherence on Roux-flasks and microcarriers. PMID- 12573003 TI - Optimization of an acoustic cell filter with a novel air-backflush system. AB - Increasing worldwide demand for mammalian cell production capacity will likely be partially satisfied by a greater use of higher volumetric productivity perfusion processes. An important additional component of any perfusion system is the cell retention device that can be based on filtration, sedimentation, and/or acoustic technologies. A common concern with these systems is that pumping and transient exposure to suboptimal medium conditions may damage the cells or influence the product quality. A novel air-backflush mode of operating an acoustic cell separator was developed in which an injection of bioreactor air downstream of the separator periodically returned the captured cells to the reactor, allowing separation to resume within 20 s. This mode of operation eliminated the need to pump the cells and allows the selection of a residence time in the separator depending on the sensitivity of the cell line. The air-backflush mode of operating a 10L acoustic separator was systematically tested at 10(7) cells/mL to define reliable ranges of operation. Consistent separation performance was obtained for wide ranges of cooling airflow rates from 0 to 15 L/min and for backflush frequencies between 10 and 40 h(-1). The separator performance was optimized at a perfusion rate of 10 L/day to obtain a maximum separation efficiency of 92 +/- 0.3%. This was achieved by increasing the power setting to 8 W and using duty cycle stop and run times of 4.5 and 45 s, respectively. Acoustic cell separation with air backflush was successfully applied over a 110 day CHO cell perfusion culture at 10(7) cells/mL and 95% viability. PMID- 12573004 TI - Comparison of batch and perfusion culture in combination with pilot-scale expanded bed purification for the production of soluble recombinant beta secretase. AB - beta-Secretase is one of the prime targets for therapeutic intervention of Alzheimer's disease. For the development of a secretase inhibitor a steady supply of large quantities of a homogeneous and active recombinant beta-secretase is a prerequisite. Therefore various culture modes were investigated using HEK-293 cells stably transfected with soluble recombinant beta-secretase. The coupling of the Fc part of human IgG1 to the ectodomain of beta-secretase (residues 1-460) allowed a fast purification of the protein with rProtA expanded bed chromatography. Batch cultures of 5 to 50 L working volume run for 7 days showed reproducible cell growth and product yields of 3 mg/L purified protein. A 20 L perfusion culture was operated for 21 days, reaching a cell density of 30 x 10(6) cells/mL at a dilution rate of 2/d. The total product yield of the perfusion culture was 1.4 g of purified protein. The effect of different perfusion rates on cell growth, protein yield, and quality was investigated and compared to the results obtained in batch cultures. Protein quality was consistent as analyzed on 1D SDS-PAGE, and the final product contained both the mature and the pro form of beta-secretase. Although the cell specific protein expression was slightly reduced in perfusion culture, a substantial increase in specific activity of over 75% was achieved. Some of the increase in activity can be explained by an increase in the percentage of the mature form of the recombinant protein. PMID- 12573005 TI - Strategies for improved dCO2 removal in large-scale fed-batch cultures. AB - Carbon dioxide buildup in large-scale reactors can be detrimental to cell growth and productivity. In case of protein X, a therapeutic glycoprotein, when cultures were scaled up from bench scale to the pilot plant, there was a 40% loss of specific productivity. The dissolved CO(2) (dCO(2)) level was 179 +/- 9 mmHg at the pilot plant scale and 68 +/- 13 mmHg at bench scale. The authors proposed a comprehensive approach to maintain dCO(2) levels between 40 and 120 mmHg throughout the 14-day fed-batch process. A cell-free experiment was used to investigate the impact of the following parameters on dCO(2) removal: (1) sparge rate, (2) agitator speed, (3) bubble size, (4) bicarbonate concentration, (5) impeller position, and (6) aeration rate at the headspace of bioreactor. dCO(2) was measured using a fiber optic based probe. dCO(2) removal rate was a strong function of sparge rate and a weak function of agitator speed. Bubble size was modulated by the presence or absence of a sparge stone (10 microm pore size, 1 cm pipe i.d.). Open pipe provided 3- to 4-fold better dCO(2) removal for the same mass transfer coefficient (k(L)a) value. A mathematical model and a bench-scale experiment indicated that the benefit of a lower level of sodium bicarbonate in the culture medium was transient for batch and fed-batch cultures. Thus, this strategy was not used at pilot scale. Decreasing top impeller position improved k(L)a of dCO(2) by 2-fold. Changing headspace aeration rate from 0.02 to 0.04 vvm had no impact on dCO(2) removal. Two pilot runs were conducted using (A) open pipe and (B) antifoam in the presence of sparge stone, both in conjunction with lower impeller position. The presence of antifoam may interfere in product purification; however, demonstration of antifoam removal can be difficult. Open pipe allowed an alternative to using antifoam, as foam level with open pipe was significantly less. Both strategies successfully reduced dCO(2) level by 2.5-fold (179 +/- 9 vs 72 +/- 9 mmHg). Titer at day 10 of culture improved by 1.5-fold. Specific productivity improved by 41%. Historically, cultures were harvested around day 9-11 because of the high amount of foam; both strategies allowed the cultures to be extended up to day 14, resulting in 2-fold higher titer compared to that of the historical control without compromising protein quality. PMID- 12573006 TI - Application of factorial design to accelerate identification of CHO growth factor requirements. AB - To accelerate recombinant CHO media and process development, we describe a simple approach to integrating multiple tasks associated with these processes including initial media design, serum-free adaptation, stability analysis and first generation scale-up. Factorial design techniques and normal probability chart representation of the results were first applied to identify potent parental CHO cell growth factors in a lean basal medium. These results were then applied to identify a suitable manufacturing medium from a panel of commercial and proprietary media formulations. When this approach was applied to recombinant CHO cell line, rapid adaptation of the cell line to an appropriate production medium occurred during culture expansion in the presence of the identified growth factor(s). This approach allows media component screening to be naturally integrated into the adaptation and scale-up processes since components that have little or no relative effect on cell proliferation are selected against as the "best" cultures are moved forward. The rapidity of the adaptation process allowed cell line stability studies to be initiated relatively early in the development process, thus providing preliminary stability information by the time the "outgrowing" culture could be scaled to 100-L reactors some 30 days after adaptation commenced. The application of full factorial design techniques allowed us to calculate the maximum number of interaction effects, the interpretation of which we believe can provide insights into growth factor biology. PMID- 12573007 TI - Enhancement of Sf-9 cell growth and longevity through supplementation of culture medium with hemolymph. AB - The benefits of insect cell culture medium supplementation with hemolymph of Lonomia obliqua were investigated. The addition of hemolymph to the medium induced high levels of cell growth, and the viability was maintained for longer periods. The maximum cell yield increased almost 3-fold after hemolymph supplementation. Cultures in their stationary phase were rescued through hemolymph supplementation, also reaching high cell concentrations. These actions were much dependent on the concentration of hemolymph; low hemolymph concentration had a positive effect in cell growth, whereas high hemolymph concentration showed a deleterious effect. Fractionation of hemolymph by gel filtration chromatography showed the presence of three factors with different activity in insect cell culture: an potential anti-apoptotic factor, a growth promoting factor, and an enzyme that hydrolyzes sucrose. Addition of hemolymph to the medium induced high levels of glucose production. The sucrose to glucose conversion was also linearly dependent upon the hemolymph concentration. Therefore, we conclude that cell growth and longevity can be increased by supplementation of the culture medium with hemolymph. PMID- 12573008 TI - Efficient lipid delivery to hybridoma culture by use of cyclodextrin in a novel granulated dry-form medium technology. AB - Lipids are critical nutrients for high density eukaryotic cell cultivation, but inclusion of lipid components into dry-form media has been technically challenging. Lipid supplements are usually supplied for separate addition after powder reconstitution and filtration, which increases manipulation and risk in a biopharmaceutical manufacturing facility. Advanced Granulation Technology (AGT) is a novel dry-form media format designed for large-scale biopharmaceutical manufacture. All components of a complex formulation are homogeneously incorporated into a single granulated medium that simply requires water addition to yield a complete reconstituted 1X medium. We investigated whether cyclodextrin technology could be combined with the AGT process to deliver usable lipid in a dry medium format. The test lipids were cholesterol and several fatty acids supplied either as an aseptic supplement to liquid media or as part of a complete AGT formulation. The test system utilized a cholesterol auxotroph, NS0, and a protein-free basal formulation (CD Hybridoma Medium), with or without supplemental lipid. Post-filtration recovery of cholesterol from AGT-processed materials compared favorably with media supplemented with liquid lipid concentrates. Cell growth and viability and expression of recombinant protein were equivalent in all test media. PMID- 12573009 TI - Cell growth arrest by nucleotides, nucleosides and bases as a tool for improved production of recombinant proteins. AB - Arresting cell growth and thus decreasing cell division potentially lessens the chance for genetic drift in the cell population; this would be of utmost importance for the consistent production of biopharmaceuticals during long periods. The drawback of the addition of well-known synchronizing agents, such as chemotherapeutics, is that they cause a disproportionate accumulation of cellular constituents, leading to cell death. The use of compounds that are naturally synthesized by the cell, as is the case of nucleotides, nucleosides, and bases (Nt/Ns/B), is shown in this work to be a promising tool. The addition of purines and pyrimidines was tested using a CHO cell line producing the secreted form of the human placental alkaline phosphatase enzyme (SEAP). From the chemical alternatives tested, AMP was the most promising compound for protein production improvement; it reduced cell growth and maintained the culture with high cell viability for long periods, while increasing SEAP specific productivity 3-fold. The use of CHO and BHK mammalian cells producing Factor VII and the use of a insect cell line (Sf9) showed that the effect of AMP addition seems to be independent of the r-protein and cell line. With the addition of AMP, accumulation of cells at the S phase was accompanied by an increase of the protein specific productivity. Addition of known synchronizing drugs (aphidicolin and doxorubicin) and application of environmental cell growth arrest strategies (depletion of nutrients and byproduct accumulation) showed also to effectively arrest CHO cell growth. A careful look onto cell cycle distribution in the different scenarios created, shows whether it is important to consider r-protein expression dependency upon cell cycle in process optimization and operation strategies. PMID- 12573010 TI - Protective effect of viral homologues of bcl-2 on hybridoma cells under apoptosis inducing conditions. AB - Targets for metabolic engineering have been identified in a hybridoma cell line to make it more robust in culture toward potential limitations inducing apoptosis. The cells were genetically modified with plasmids harboring endogenous bcl-2 gene and also with viral Bcl-2 homologues, particularly ksbcl-2 and bhrf-1 genes. When cells were exposed to apoptosis-inducing conditions (i.e., glutamine free medium), the control cells exhibited a decrease in viable cell number within the first 12 h, whereas, for the bcl-2 and ksbcl-2 transfected cell cultures, the viable cell number did not exhibit any clear decrease until after 60 h. Furthermore, hybridoma cells expressing the viral homologue bhrf-1 were even more resistant to cell death, showing a decrease in viability of only 50% at 72 h of culture in glutamine-deprived medium, substantially lower than the 90% viability decrease observed for the control culture. In addition, and most relevant for further bioprocess applications, the cells genetically modified could be brought back to growth conditions even after being exposed to glutamine-deprived conditions during a significant time window, up to 72 h. PMID- 12573012 TI - Metabolic screening of mammalian cell cultures using well-plates. AB - For use in a broad spectrum of cell culture applications, we have devised a novel method, termed High-Throughput Metabolic Screening (HTMS), with which to more rapidly screen the overall activity of major metabolic pathways of mammalian cells. This current protocol uses adaptations of theoretical and experimental techniques from metabolic and cell culture engineering. First, HTMS makes use of a simplified metabolic network for metabolic flux analysis. Despite its simplicity, the network is capable of generating flux distributions and ATP production rates that are comparable to a more detailed network. Second, HTMS makes use of microtiter well-plate technology and adaptations of well-known enzymatic assays to increase precision and throughput for cell culture experiments. Multireplicate, multiparallel cultures in the sub-milliliter scale yield very precise metabolic rates using common laboratory equipment and at a fraction of the cost and time of traditional experiments with T-flasks, spinner flasks, or bioreactor systems. The simplicity of the network and the well-plate assays synergistically comprise a new, extremely useful, broadly applicable, and relatively inexpensive way to probe cell cultures for metabolic effects, screen drugs and toxins, optimize media, and support the development of bioprocesses. The simplified network and cell culture and analytical assays are also useful for undergraduate, graduate, and professional training. PMID- 12573011 TI - Improving glucose and glutamine metabolism of human HEK 293 and Trichoplusia ni insect cells engineered to express a cytosolic pyruvate carboxylase enzyme. AB - Metabolic engineering has been defined as a directed improvement of product formation or cellular properties by modification of specific biochemical pathways or introduction of new enzymatic reactions by recombinant DNA technology. The use of metabolic flux analysis (MFA) has helped in the understanding of the key limitation in the metabolic pathways of cultured animal cells. The MFA of the major nutrients glucose and glutamine showed that the flux of glucose to the TCA cycle and its subsequent utilization is limited as a result of the lack of certain key enzymes in the pathway. One of the key enzymes controlling this flux is pyruvate carboxylase. Introduction of this enzyme into mammalian cells has been shown to improve the utilization of glucose and limit the production of lactate and ammonia, which are deleterious to cell growth. In the present work a yeast pyruvate carboxylase gene has been introduced into mammalian (HEK 293) and insect (Trichoplusia ni High-Five) cells, resulting in the cytosolic expression of the enzyme. In both cases the resulting transfected cells were able to utilize glucose and glutamine more efficiently and produce lower amounts of lactate and ammonia. Differences in the amino acid utilization pattern were also observed, indicating changes in the basic metabolism of the cells. The performance of the transfected cells as expression systems for adenovirus and baculovirus vectors, respectively, has also been examined. The results obtained and their impact on the process development for protein and viral vector production are discussed. PMID- 12573013 TI - Toward higher order control modalities in mammalian cells-independent adjustment of two different gene activities. AB - Heterologous higher order control modalities will be important tools for targeted multigene interventions in next-generation gene therapy, tissue engineering, and sophisticated gene-function studies. In this study, we present the design and rigorous quantitative analysis of a variety of different dual-regulated gene transcription control configurations combining streptogramin- and tetracycline responsive expression systems in a one-vector format. Quantitative assessment of dual-regulated expression performance in various mammalian and human cell lines is based on two compatible secreted reporter genes, SEAP, the human placental secreted alkaline phosphatase, and the recently developed SAMY, the secreted alpha-amylase. Assembly of streptogramin- and tetracycline-responsive transgene control units in consecutive (--> -->), divergent (<-- -->), and convergent (--> <--) orientation showed excellent regulation characteristics in most genetic arrangements exemplified by neglectable interference and high transgene induction ratios in all four control settings (ON/ON, OFF/ON, ON/OFF, OFF/OFF). The overall regulation performance of divergent dual-regulated expression configurations could be substantially increased when placing noncoding stuffer fragments or insulator modules between the divergently oriented antibiotic-responsive promoters. Dual-regulated expression technology pioneers artificial higher order gene control networks that will likely enable new opportunities in multigene metabolic engineering and generate significant therapeutic impact. PMID- 12573014 TI - Engineering mRNA translation initiation to enhance transient gene expression in chinese hamster ovary cells. AB - To increase transient expression of recombinant proteins in Chinese hamster ovary cells, we have engineered their protein synthetic capacity by directed manipulation of mRNA translation initiation. To control this process we constructed a nonphosphorylatable Ser(51)Ala site-directed mutant of eIF2alpha, a subunit of the trimeric eIF2 complex that is implicated in regulation of the global rate of mRNA translation initiation in eukaryotic cells. Phosphorylation of eIF2alpha by protein kinases inhibits eIF2 activity and is known to increase as cells perceive a range of stress conditions. Using single- and dual-gene plasmids introduced into CHO cells by electroporation, we found that transient expression of the eIF2alpha Ser(51)Ala mutant with firefly luciferase resulted in a 3-fold increase in reporter activity, relative to cells transfected with reporter only. This effect was maintained in transfected cells for at least 48 h after transfection. Expression of the wild-type eIF2alpha protein had no such effect. Elevated luciferase activity was associated with a reduction in the level of eIF2alpha phosphorylation in cells transfected with the mutant eIF2alpha construct. Transfection of CHO cells with the luciferase-only construct resulted in a marked decrease in the global rate of protein synthesis in the whole cell population 6 h post-transfection. However, expression of the mutant Ser(51)Ala or wild-type eIF2alpha proteins restored the rate of protein synthesis in transfected cells to a level equivalent to or exceeding that of control cells. Associated with this, entry of plasmid DNA into cells during electroporation was visualized by confocal microscopy using a rhodamine-labeled plasmid construct expressing green fluorescent protein. Six hours after transfection, plasmid DNA was present in all cells, albeit to a variable extent. These data suggest that entry of naked DNA into the cell itself functions to inhibit protein synthesis by signaling mechanisms affecting control of mRNA translation by eIF2. This work therefore forms the basis of a rational strategy to generically up-regulate transient expression of recombinant proteins by simultaneous host cell engineering. PMID- 12573015 TI - Exploring vaccinia virus as a tool for large-scale recombinant protein expression. AB - A recombinant vaccinia virus was engineered to express enhanced green fluorescent protein (EGFP) under control of the T7 promoter using the VOTE expression system in HeLa cells. Infection of HeLa cells with this virus and induction with IPTG demonstrated the utility of this construct for easily measuring protein expression. This construct was used to evaluate several production parameters, specifically, multiplicity of infection (MOI), volume during infection, and serum concentration during the infection phase. In static culture, increasing multiplicity of infection was found to increase expression of EGFP up to a plateau around MOI of 1.0. Expression was also shown to increase with decreasing volume during the infection phase. Serum concentration during the infection phase was only marginally significant from 0 to 7.5%. Cytodex 3 microcarriers were found to have the best characteristics for HeLa cell growth. These cells were grown and infected in microcarrier spinner flask culture, and the maximum expression was 2.2 microg EGFP/(million cells at the time of infection), demonstrating the ability of this system to successfully express recombinant proteins at larger scale. PMID- 12573016 TI - Development of a Chinese hamster ovary cell line for recombinant adenovirus mediated gene expression. AB - Recombinant human adenovirus (rhAd) has been used extensively for functional protein expression in mammalian cells including those of human and nonhuman origin. High-level protein production by rhAd vectors is expected in their permissive host cells, such as the human embryonic kidney 293 (HEK293) cell line. This is attributed primarily to the permissiveness of HEK293 to rhAd infection and their ability to support viral DNA replication by providing the missing El proteins. However, the HEK293 cells tend to suffer from cytopathic effect (CPE) as a result of virus replication. Under these circumstances, the host cell function is compromised and the culture viability will be reduced. Consequently, newly synthesized polypeptides may not be processed properly at posttranslational levels. Therefore, the usefulness of HEK293 cells for the expression of complex targets such as secreted proteins could be limited. In the search for a more robust cell line as a production host for rhAd expression vectors, a series of screening experiments was performed to isolate clones from Chinese hamster ovary K1 (CHO-K1) cells. First, multiple rounds of infection of CHO-K1 cells were performed utilizing an rhAd expressing GFP. After each cycle of infection, a small population of CHO cells with high GFP levels was enriched by FACS. Second, individual clones more permissive to human adenovirus infection were isolated from the highly enriched subpopulation by serial dilution. A single clone, designated CHO-K1-C5, was found to be particularly permissive to rhAd infection than the parental pool and has served as a production host in the successful expression of several secreted proteins. PMID- 12573018 TI - Quantitative monitoring for secreted production of human interleukin-2 in stable insect Drosophila S2 cells using a green fluorescent protein fusion partner. AB - Human interleukin-2 (hIL-2) production in Escherichia coli and insect cell/baculovirus expression systems can be inefficient. Here we investigated secreted production of hIL-2 fused with green fluorescent protein (GFP) as a versatile fusion partner in optimized stably transfected insect Drosophila melanogaster S2 cells. This nonlytic S2 insect cell expression system employs a plasmid vector and allows for secretion of functional human proteins. We report that, following stable transfection and induction, S2 cells secreted hIL-2 as a fusion protein (approximately 2.3 microg/mL yield), with a secretion efficiency of approximately 90%. Regression analysis indicated a single linear relationship existed between GFP fluorescence and hIL-2 mass in both whole cell and secreted medium samples, indicating that in vivo monitoring and quantification of target foreign protein expression and even secretion is possible using this system. The simple comparative measurement of GFP fluorescence also allowed monitoring of secretion efficiency during periods of high GFP/hIL-2 expression. PMID- 12573017 TI - Epstein-Barr virus vectors provide prolonged robust factor IX expression in mice. AB - We demonstrate that vectors incorporating components from Epstein-Barr virus (EBV) for retention and from human genomic DNA for replication greatly enhance the level and duration of marker gene expression in dividing cultured cells. The same types of vectors were tested in vivo by high-pressure tail vein injection of naked DNA in mice, resulting in liver delivery and expression. The therapeutic gene was a human factor IX (hFIX) minigene comprising genomically derived 5', 3', and intronic sequences that provided relatively good gene expression in vivo. We demonstrated that addition of the EBV EBNA1 gene and its family of repeats binding sites provided a 10- to 100-fold increase in prolonged hFIX expression in mouse liver. A single 25-microg dose of vector DNA generated normal (>5 microg/mL) levels of hFIX throughout the 8 month duration of the experiment. Vector DNA with or without the EBV sequences was retained in liver cells, and vector replication was not a factor in these nondividing liver cells. Instead, it appears that enhancement of stable hFIX expression by the EBV components was responsible for the increased level and duration of therapeutic gene expression. The EBV sequences also significantly enhanced stable expression of a vector carrying the full genomic hFIX gene delivered to mouse liver. These results underline the crucial importance of appropriate gene expression signals on gene therapy vectors and the utility of EBV sequences in particular for increasing stable gene expression. PMID- 12573019 TI - Improvement of monoclonal antibody production in hybridoma cells by dimethyl sulfoxide. AB - Hybridoma cultures are routinely used as a source for monoclonal antibody (mAb) production necessary for preclinical evaluation. However, these cultures typically have low volumetric and specific productivities. In this article, we examined the use and the timing of addition of dimethyl sulfoxide (DMSO) as a medium additive to improve mAb production in our hybridoma clone 19 (c19) cultures. From shake flask studies, we defined the optimal DMSO concentration and time of addition for improved productivity. This timing coordinated with high cell viability and density. Hybridoma cultures treated with DMSO up to 0.3% (v/v) possessed cell densities and viabilities comparable to untreated control. We demonstrated that 0.2% (v/v) DMSO added to shake flask cultures at their maximal viable cell densities resulted in a 2-fold increase in specific mAb production. This procedure was scaleable up to 20 L Cellbags (Wave Bioreactors) with similar titer improvement. Moreover, DMSO treatment did not affect the bioactivity or glycosylation profiles of the mAb. PMID- 12573020 TI - High-level expression of recombinant IgG in the human cell line per.c6. AB - The number of therapeutic monoclonal antibodies in production is expected to rise rapidly in the next few years. As a result, there is much focus on the optimization of antibody expression platforms. Several issues are important including the speed of transition from bench to manufacturing, yield of IgG, and quality (particularly of the glycan structures present on immunoglobulins). We have characterized the human cell line PER.C6 for its ability to produce recombinant IgG. Production yields are still being optimized, but in nonfed batch culture, PER.C6 is able to grow to a cell density of 5 x 10(6) cells/mL and produce 300-500 mg/L IgG; this is likely to increase significantly in fed batch cultures. The generation of antibody-producing cell lines is fast, as rounds of amplification of inserted genes are not required for high production yields. The gene copy number of inserted genes is in the region of 1-10 copies per genome. In addition, PER.C6 is a human cell line, and so does not add glycans, which are immunogenic in humans. A core fucose molecule is essentially always present, and galactose residues are present at a physiological level (0, 1, and 2 galactose residues per glycan are present at a ratio of 1:2:1). No hybrid or high-mannose structures are seen. PMID- 12573021 TI - Enhancement of monoclonal antibody production by lysine-containing peptides. AB - In the search for peptides that could effectively enhance the monoclonal antibody production of a model hybridoma, the performance of five lysine-containing peptides was compared. The capacity of the peptides to enhance the monoclonal antibody yield correlated with their growth-suppressing activity. No correlation of the production-enhancing activity with the character of the distribution of cell-cycle phases could be found. All of the tested peptides, including the negative control peptide Gly-Phe-Gly, altered the cell-cycle phases distribution in favor of the proportion of the S phase. The peptides added to the hybridoma culture were found to be gradually decomposed into dipeptides and free amino acids. Among the set of tested lysine-containing di- to pentapeptides, the best results were obtained with the tripeptide Gly-Lys-Gly. The growth-suppressing and production-enhancing capacity of this peptide supplement was obviously associated with the temporary presence of the intact peptide molecule in the culture media, because the addition of a mixture of free amino acids constituting this peptide, i.e., glycine and lysine, displayed a different effect-a slight promotion of cell growth. PMID- 12573022 TI - Improved fermentation processes for NS0 cell lines expressing human antibodies and glutamine synthetase. AB - To meet the increasing requirement for therapeutic antibodies to conduct clinical trials, an enhanced culture medium and fed-batch process was developed for GS-NS0 cell lines. This process was shown to produce high concentrations of monoclonal antibodies for several cell lines expressing different antibodies. Cells were adapted to growth in a glutamine- and serum-free medium containing bovine serum albumin (BSA), cholesterol, and transferrin. A number of amino acids were found to be depleted during cell culture. The concentrations of these amino acids were increased, and further cell culture analyses were performed. This process of cell growth and analysis was repeated over multiple cycles until no depletion was detected. This resulted in an amino acid supplement that was shown to be generic and enhanced antibody productivity up to 5-fold for the three cell lines tested. Transferrin was replaced using tropolone, a lipophilic iron chelator and ferric ammonium citrate. Cell growth was equivalent to that in transferrin-containing medium over the wide ranges tested. A concentrated feed solution, based on the amino acid supplement and the components of the serum- and protein-free supplements, was formulated. Addition of this feed in response to metabolic requirements resulted in a harvest titer a further 2-fold higher than the enhanced culture medium. Harvest antibody titers of up to 600 mg/L were achieved for three cell lines expressing different antibodies, representing an increase of 10-fold over the starting concentrations. PMID- 12573023 TI - Effect of doxycycline-regulated ERp57 expression on specific thrombopoietin productivity of recombinant CHO cells. AB - In an attempt to increase the specific thrombopoietin (TPO) productivity (q(TPO)) of recombinant Chinese hamster ovary (rCHO) cells (TPO-33), the effect of expression level of ERp57, an isoform of protein disulfide isomerase, on q(TPO) was investigated. To regulate ERp57 expression level, the Tet-Off system was first introduced in TPO-33 cells and stable Tet-Off cells (TPO-33-Tet-Off) were screened by the luciferase assay. The rCHO cells with a doxycycline-regulated ERp57 expression system (TPO-33-ERp57) were obtained by cotransfection of pTRE ERp57 and pTK-Hyg expression vectors into TPO-33-Tet-Off cells and subsequent screening by Western blot analysis of ERp57 and an enzyme-linked immunosorbent assay of secreted TPO. Western blot analysis showed that ERp57 expression level in TPO-33-ERp57 cells could be regulated tightly by the addition of different concentrations of doxycycline to a culture medium. A doxycycline concentration of 1 microg/mL, which did not influence cell growth and TPO production of TPO-33-Tet Off cells, was high enough to suppress the ERp57 expression to a basal level. Compared with the basal level, a 1.7-fold increase in ERp57 expression level was obtained in the absence of doxycycline. This increased expression level of ERp57 resulted in a 2.1-fold increase in q(TPO) without growth inhibition, probably as a result of the chaperone-like activity of ERp57 in CHO cells. Taken together, the results obtained here demonstrate that q(TPO) of rCHO cells can be increased by elevating the expression level of ERp57. PMID- 12573024 TI - Novel insect cell line capable of complex N-glycosylation and sialylation of recombinant proteins. AB - Paucimannose or oligomannose structures are usually attached to glycoproteins produced by insect cells, while mammalian glycoproteins usually have complex glycans. The lack of complex glycosylation has limited the use of the insect cell baculovirus expression vector system (BEVS), despite its high productivity and versatility. The availability of cell lines capable of complex glycosylation can overcome such a problem and potentially increase the utility of BEVS. In this work the capability of two novel cell lines, one from Pseudaletia unipuncta (A7S) and one from Danaus plexippus (DpN1), to produce and glycosylate a recombinant protein (secreted human placental alkaline phosphatase, SeAP) was assessed. SeAP produced by Tn5B1-4 cells at a low passage number (<200) was utilized for comparison. The optimal conditions for the production of SeAP by DpN1 cells were defined, and the glycosylation profiles of SeAP produced by the cell lines were quantitatively determined. Both the A7S and the DpN1 cells produced lower concentrations of SeAP than the Tn5B1-4 cells. Less than 5% of the glycans attached to SeAP produced by the Tn5B1-4 cells had complex forms. Glycans attached to SeAP from A7S cells contained 4% hybrid and 8% complex forms. Galactosylated biantennary structures were identified. Glycans attached to SeAP produced by the DpN1 cell line had 6% hybrid and 26% complex forms. Of the complex forms in SeAP from DpN1, 13% were identified as sialylated glycans. The galactosyltransferase activity of the three cell lines was measured and correlated to their ability to produce complex forms. Even though neither novel cell line produced as much recombinant protein as the Tn5B1-4 cells, the glycosylation of SeAP expressed by both cell lines was more complete. These novel cell lines represent interesting alternatives for the production of complex glycosylated proteins utilizing the BEVS. PMID- 12573025 TI - Production of a sialylated N-linked glycoprotein in insect cells: role of glycosidases and effect of harvest time on glycosylation. AB - Using a nonengineered Trichoplusia ni insect cell line, Tn-4s, infected with an Autographa californica recombinant baculovirus, 20% sialylation of human secreted placental alkaline phosphatase (SEAP) was observed. In contrast to this level of sialylation, intermediate complex forms with terminal galactose or N acetylglucosamine were found in low proportions (<3% and <1%, respectively). We tested whether time of harvest or degradation of intermediate complex forms is responsible for this distribution of glycoforms. Spinner-flask cultures were infected with the SEAP baculovirus expression vector, and the cultures were harvested 48, 72, and 96 h post-infection. Structural analysis revealed that the glycoform distribution of SEAP was very similar at the different times of harvest, indicating that the cellular machinery was not significantly affected by the progress of infection and that the glycoforms obtained were stable. High levels of beta-galactosidase and N-acetylglucosaminidase activity were detected throughout infection. In contrast, sialidase activity was below detection level both in cell extracts and in supernatants. These levels of glycosidases activities raise the possibility that intermediate complex glycoforms may be degraded while sialylated forms should not experience significant degradation in this cell line. However, culture in the presence of extracellular beta galactosidase and N-acetylglucosaminidase inhibitors did not significantly improve glycosylation, suggesting that extracellular degradation processes are not taking place. Instead, results suggest that the intracellular machinery of the Tn-4s cells tends to either shunt the glycans to paucimannosidic forms or drive them completely to sialylation. PMID- 12573026 TI - PH, pCO2, and temperature effect on R-adenovirus production. AB - The effects of pH, carbon dioxide vapor pressure, pCO(2), and temperature on E1 and E3 deleted recombinant adenovirus vector (rAV) production with HEK293S cells have been studied in the ranges of pH = 6.7-7.7, pCO(2) = 0.05-0.20 atm, and T = 32-39 degrees C, respectively. The experiments were performed in four 500-mL bioreactors in parallel, which make possible the reduction of inter-run variability. Cell concentration and viability, relative oxygen uptake rate (OUR), fluorescence, and viral titer were measured. It was found that, although pH and pCO(2) did not affect significantly cell viability in the range studied, they had an important effect on virus titer. pCO(2) allowed the maximum production of rAV at 0.05 atm, and pH showed a very sharp optimum at 7.2. Temperature had an effect on both cell metabolism and virus titer. Low temperature prolonged cell viability and high OUR. Most of all, a 3-fold increase in virus yield was found at 35 degrees C compared to that at 37 degrees C, while 32 degrees C was not as beneficial (1.5-fold increase). This finding could have an important impact on large-scale production. This phenomenon was modeled using a simple 3-parameter synthesis-decay model. This model shows how the optimum gain in virus production at 35 degrees C is due to a balance between the production and decay processes at that temperature. PMID- 12573027 TI - Impact of process conditions on the centrifugal recovery of a disabled herpes simplex virus. AB - Despite continuous improvements in culturing and recovery techniques, high-titer stocks of purified disabled herpes simplex virus type-1 (HSV-1 DIS) vector for drug discovery and use in preclinical and clinical trials are currently difficult to achieve. Efforts to improve their centrifugal recovery have been addressed in this paper. The operation of a swing-out centrifuge rotor was assessed, and its operational conditions were defined for the recovery of viable HSV-1 DIS. 80% virus recovery was achieved after 90 min at 26000g. The 20% loss of virus was attributed to damage to the viral envelope by overcompaction of the pellet and impaction with the base of the centrifuge tube. Virus recovery was increased by a further 10% by using a fixed-angle centrifuge rotor operating at 26000g. Plaque assays of recovered HSV-1 DIS gave values on the order of 10(6) pfu/mL, compared to values typically above 10(9) pfu/mL obtained for the replication-competent HSV 1 viron. PMID- 12573028 TI - Small increases in pH enhance retroviral vector transduction efficiency of NIH 3T3 cells. AB - Increases in pH between 7.1 and 7.7 increase the efficiency of polybrene (Pb)- and protamine sulfate (PS)-aided retroviral transduction of NIH-3T3 cells in a serum-lot-dependent manner. The increase in Pb-aided transduction efficiency at pH 7.7, relative to the value at pH 7.33, ranged from 13% to 49% for three serum lots. For a constant Moloney murine leukemia virus (MMLV) vector dilution at pH 7.33, three different serum lots resulted in absolute transduction efficiencies ranging from 29% to 53% using Pb. At the same vector dilution, PS-aided transduction was less effective on an absolute basis than Pb-aided transduction, but the benefit of elevated pH was more pronounced with PS. There was a similar enhancement with PS at elevated pH for a murine stem cell virus (MSCV) vector as for the MMLV vector. The benefit at pH 7.7 for PS-aided transduction was partially due to greater PS stability at elevated pH. Heat inactivating the serum supplement or adding protease inhibitors helped to stabilize PS. This increased the absolute transduction efficiency but decreased the relative benefit of elevated pH to a level similar to that for Pb-aided transduction. Incubating Pb with the vector at pH 7.1 for 10 min, prior to readjusting to pH 7.7 and transducing the cells, was sufficient to abrogate the beneficial effects of transduction at pH 7.7. In contrast, prior exposure of PS with vector at pH 7.1 did not affect subsequent transduction at pH 7.7. These results indicate that pH is an important variable in retroviral transduction and that the relative benefits of Pb or PS on retroviral vector transduction will vary with the pH, polymer addition method, and serum lot. PMID- 12573029 TI - Modulation of cell cycle for enhancement of antibody productivity in perfusion culture of NS0 cells. AB - A prolonged period of high productivity at high cell density is desirable for industrial production of biopharmaceuticals. Previous efforts have shown that cessation of cell proliferation in low cell density culture results in increased productivity. We report here further results on multigenic manipulation of cell cycle and apoptosis to enhance productivity at high cell density. The NS0 6A1/4 9F myeloma cell line, which constitutively expresses a chimeric IgG4 antibody and inducibly expresses the p21(CIP1) cyclin-dependent kinase inhibitor has been further engineered to constitutively overexpress the Y28 mutant Bcl-2 anti apoptotic protein. The effects of overexpression of p21(CIP1) and Bcl-2 on cell proliferation, cell viability, and antibody production has been investigated in batch and continuous perfusion cultures. In both cultures the p21(CIP1) protein arrested cell proliferation, confirming the previous results in low-density culture of 4-fold increase in antibody production, whereas mutant Bcl-2 expression has not resulted in any significant improvement in cell viability of arrested cells. This study demonstrates that it is possible to enhance the productivity of relatively high-density continuous mammalian cell cultures by arresting the cell cycle in G1 phase. PMID- 12573031 TI - Stability of recombinant protein production in the GS-NS0 expression system is unaffected by cryopreservation. AB - Mammalian cells form a very important part of the repertoire of production systems available to scientists involved in the production of recombinant proteins. During the production of therapeutic proteins it is vital for regulatory approval of products that no phenotypic or genetic changes are observed in the cell line or product. As part of the generation and development of therapeutic protein production, cell lines have to be frozen at various stages to create cell banks. If cryopreservation and revival of frozen stocks were to give rise to any phenotypic changes in the cells, this would again be detrimental to the further development of that particular cell line. This study uses one of the most industrially important expression systems, the GS-NS0 expression system, to examine the effect of cryopreservation on the growth and productivity profile of cell lines that exhibit differential degrees of stability during prolonged (production) culture periods. Results show that cryopreservation and revival procedures do not alter the stability characteristics of cell lines. This type of information is of great value in definition of protocols for cell line development. PMID- 12573030 TI - Versatile expression system for rapid and stable production of recombinant proteins. AB - Previously we reported the development of a novel expression system with Tat/TAR oriP vectors and HKB11 cell line, which supports high level protein expression (Cho et al. Cytotechnology 2001, 37, 23-30). In the present study, we further demonstrated that HKB11 cells are suitable for high throughput expression (microgram scale) of genomic candidates in transient transfection system for in vitro evaluation of biological functions. HKB11 cells were also shown to support the production of milligram to gram quantities of protein drug candidates for in vivo evaluation of efficacy in various disease models. Stable HKB11 clones secreting high levels of a tissue factor (TF; 40-50 pg/c/d) and B-domain deleted recombinant factor VIII (BDDrFVIII; 5-10 microU/c/d) were derived under serum free conditions. The specific productivity for these two proteins from the HKB11 cells was 10-fold greater than those from CHO cells derived under the similar conditions. In conclusion, we have demonstrated that the HKB11 cell line is well suited for transient and long-term production of recombinant proteins. PMID- 12573032 TI - Effect of time of harvest of budded virus on the selection of baculovirus FP mutants in cell culture. AB - Rapid formation and selection of FP (few polyhedra) mutants occurs during serial passaging of Helicoverpa armigera nucleopolyhedrovirus (HaSNPV) in insect cell culture. The production of HaSNPV for use as biopesticides requires the passaging of the virus over a number of passages to produce enough virus inoculum for large scale fermentation. During serial passaging in cell culture, FP mutants were rapidly selected, resulting in declined productivity and reduced potency of virus. Budded virus (BV) is usually harvested between 72 and 96 h postinfection (hpi) in order to obtain a high titer virus stock. In this study, the effect of time of harvest (TOH) for BV on the selection rate of HaSNPV FP mutants during serial passaging was investigated. BV were harvested at different times postinfection, and each series was serially passaged for six passages. The productivity and percentage of FP mutants at each passage were determined. It was found that the selection of FP mutants can be reduced by employing an earlier TOH for BV. Serial passaging with BV harvested at 48 hpi showed a slower accumulation of FP mutants compared to that of BV harvested after 48 hpi. Higher cell specific yields were also maintained when BV were harvested at 48 hpi. When BV that were formed between 48 and 96 hpi were harvested and serially passaged, FP mutants quickly dominated the virus population. This suggests that the BV formed and released between 48 and 96 hpi are most likely from FP mutant infected cells. PMID- 12573033 TI - The role of an evolutionarily conserved cis-proline in the thioredoxin-like domain of human class Alpha glutathione transferase A1-1. AB - The thioredoxin-like fold has a betaalphabetaalphabetabetaalpha topology, and most proteins/domains with this fold have a topologically conserved cis -proline residue at the N-terminus of beta-strand 3. This residue plays an important role in the catalytic function and stability of thioredoxin-like proteins, but is reported not to contribute towards the stability of glutathione S-transferases (GSTs) [Allocati, Casalone, Masulli, Caccarelli, Carletti, Parker and Di Ilio (1999) FEBS Lett. 445, 347-350]. In order to further address the role of the cis proline in the structure, function and stability of GSTs, cis -Pro-56 in human GST (hGST) A1-1 was replaced with a glycine, and the properties of the P56G mutant were compared with those of the wild-type protein. Not only was the catalytic function of the mutant dramatically reduced, so was its conformational stability, as indicated by equilibrium unfolding and unfolding kinetics experiments with urea as denaturant. These findings are discussed in the context of other thioredoxin-like proteins. PMID- 12573034 TI - High-mobility group A1 proteins are overexpressed in human leukaemias. AB - High-mobility group A (HMGA) proteins are non-histone nuclear proteins that bind DNA and several transcription factors. They are involved in the regulation of chromatin structure and function. HMGA protein expression is low in normal adult tissues, but abundant during embryonic development and in several human tumours. Rearrangements of the HMGA genes have been frequently detected in human benign tumours of mesenchymal origin, e.g. lipomas, lung hamartomas and uterine leiomiomas. HMGA proteins have been implicated in the control of cell growth and differentiation of the pre-adipocytic cell line 3T3-L1. In an attempt to better understand the role of HMGA1 proteins in haematological neoplasias and in the differentiation of haematopietic cells, we have investigated their expression in human leukaemias and in leukaemic cell lines induced to terminal differentiation. Here we report HMGA1 overexpression in most fresh human leukaemias of different origin and in several leukaemic cell lines. Moreover, differentiation of three cell lines towards the megakaryocytic phenotype was associated with HMGA1 protein induction, whereas induction of erythroid and monocytic differentiation generally resulted in reduced HMGA1 expression. PMID- 12573035 TI - Panel discussion: current issues in the prevention and management of surgical site infection--part 1. PMID- 12573036 TI - Surgical site infections: epidemiology and prevention. AB - BACKGROUND: The overall incidence of surgical site infection (SSI) has been estimated to be 2.8% in the United States, according to the U.S. Centers for Disease Control and Prevention, although the data may underrepresent the true incidence of such infections owing to inherent problems with voluntary self reporting by surgeons of infections that occur in the ambulatory surgical setting. This review analyzes the reasons why patients are at risk and what can be done to minimize the risk. METHODS: Review of the pertinent English-language literature. RESULTS: Factors that contribute to the development of SSI include those that arise from the patient's health status, those that relate to the physical environment where surgical care is provided, and those that result from clinical interventions that increase the patient's inherent risk. Careful patient selection and preparation, including the judicious use of antibiotic prophylaxis, can decrease the overall risk of infection, especially following clean contaminated and contaminated operations. However, antibiotics are not a substitute for attention to detail and meticulous surgical technique, and can increase the risk of nosocomial infection following injudicious use (that is, overuse). CONCLUSION: Most SSIs can be attributed to risk factors inherent to the patient, rather than to inherently flawed surgical care. Nonetheless, the surgeon can minimize the risk to the patient through careful patient selection and preparation, attention to technical details and awareness of the operating room environment, and the selective use of short-duration, narrow-spectrum antibiotic prophylaxis for appropriate patients. PMID- 12573037 TI - Cellular, biochemical, and clinical aspects of wound healing. AB - BACKGROUND: The response to tissue injury requires the symphonious interaction of immune cells, keratinocytes, fibroblasts, and endothelial cells, which unite to regenerate the damaged epithelium. Recent insights have elucidated the cellular and molecular mechanisms required for wound healing and have raised the prospect of novel therapeutic targets. METHODS: Review of the pertinent literature. RESULTS: The initial inflammatory response leads to the influx of macrophages and neutrophils, which release cytokines, growth factors, and nitric oxide, and induce nearby keratinocytes to migrate across the wounded epithelium. This process, known as re-epithelialization, requires integrin-mediated activation of Rho-GTPases. The subsequent influx of fibroblasts and endothelial cells results in the production of tissue stroma and formation of new blood vessels, which lead to the generation of functional tissue. Importantly, disease states associated with impaired or excessive wound healing can be attributed to defects in these responses, providing a rationale for the use of evidence-based biological therapies. CONCLUSION: The elucidation of the cellular and biochemical response to wound healing is essential for an understanding to the treatment of clinical conditions during which impaired healing is encountered. PMID- 12573038 TI - The economic costs of surgical site infection. AB - BACKGROUND: Surgical site infections (SSIs) remain common and are a major cause of postoperative morbidity. Less well recognized is the cost of these complications, both in the direct costs of care and also in terms of lost economic productivity when workers are disabled as a result of an infection. This review was undertaken to bring into focus the relevant literature regarding the costs associated with SSI. METHODS: Review of the pertinent English language literature. RESULTS: The estimated costs of SSI vary as a function of locale, type of operation, and the extent (depth) of the infection itself. Superficial SSIs cared for in the National Health Service of the United Kingdom have an estimated cost of less that 400 dollars/case, whereas estimates range into the tens of thousands of dollars per case in the United States for complex infections such as infected joint prostheses or sternal infections following cardiac surgery. The magnitude of the problem is emphasized by a report from Denmark, which shows that the cost of care for surgical site infections consumes 0.5% of the annual hospital budget. CONCLUSION: SSI is morbid and expensive. However, not all SSIs are the same. Reports of economic costs that do not stratify for the depth of the infection or the context in which the infection occurs are potentially misstating the magnitude of these complications. PMID- 12573040 TI - Literature-based evaluation of the potential risks associated with impregnation of medical devices and implants with triclosan. AB - BACKGROUND: This report is a review of the published literature for studies of triclosan that address mechanism of action, efficacy on skin and in the oral cavity, and the potential for development of resistance. METHODS: Triclosan citations from the past three decades were searched using Medline and other search engines. The techniques used in these studies included in vitro antimicrobial sensitivity, molecular genetics, and enzyme and membrane biochemistry. Oral cavity efficacy and resistance studies were conducted in human volunteers in trials lasting up to 7 months. Efficacy on skin was reported in clinical trials lasting up to 12 months. RESULTS: The minimal inhibitory concentration of triclosan against Staphylococcus aureus and Escherichia coli is reported to be 0.1 and 5.0 microg/mL, respectively. Triclosan acts by blocking enoyl acyl carrier protein reductase, an enzyme essential for fatty acid biosynthesis. Its biocidal activity involves a plethora of nonspecific perturbations of cellular structural elements, including the cell membrane. In the oral cavity, triclosan use was associated with significant reductions in recoverable flora; there was no evidence of resistance or emergence of opportunistic pathogens. On skin, in a neonatal intensive care unit, triclosan use was associated with a significant reduction in methicillin-resistant S. aureus (MRSA) infections, a diminished need for antibiotics, and a decreased incidence of nosocomial infections. CONCLUSION: There is currently no evidence that long-term application of triclosan products to the skin or oral cavity selects for triclosan-resistant populations. Given the short-term nature of suture use, it is highly unlikely that such use would do other than reduce the risks of postoperative infection. PMID- 12573039 TI - Chemistry and safety of triclosan, and its use as an antimicrobial coating on Coated VICRYL* Plus Antibacterial Suture (coated polyglactin 910 suture with triclosan). AB - BACKGROUND: The safety of the antimicrobial agent triclosan was reviewed, and the biocompatibility of coated polyglactin 910 suture with triclosan was evaluated. METHODS: Acute single exposure LD(50) values for triclosan were determined in multiple species by several routes of administration. Subacute to chronic toxicity for dermal and oral exposure to triclosan was determined in multiple species in studies of up to one year's duration. Chronic oral toxicity/carcinogenicity potential was determined in 2-year studies in rodents and non-rodents. The genotoxicity potential of triclosan was determined using a battery of standard assays. Reproductive toxicity and teratology studies were conducted in rodents and non-rodents. Immunotoxicity studies in guinea pigs and the repeat-insult patch test in humans were conducted to assess the potential for sensitization reactions. Pharmacokinetic studies were conducted in animals and humans to understand the metabolic profile of triclosan. Preclinical biocompatibility studies conducted on coated polyglactin 910 suture with triclosan included in vitro cytotoxicity, in vivo intracutaneous reactivity, material-mediated pyrogenicity, and intramuscular tissue reaction/absorption studies. RESULTS: The oral LD(50) values for triclosan ranged from 3,750 to 5,000 mg/kg, whereas the LD(50) after subcutaneous injection was >14,600 mg/kg. Safety factors calculated from repeated daily dosing studies ranged from 1,000 to 25,000 times the no-observed-effect levels. There was no evidence of carcinogenic potential in either species, and genotoxicity studies were negative. Reproductive toxicity studies did not reveal any evidence of teratogenic potential. There was no evidence of skin sensitization potential in controlled studies. Pharmacokinetic studies in animals and humans have shown that triclosan is rapidly absorbed, well distributed in the body, metabolized in the liver, and excreted by the kidneys, with no indication of accumulation over time. Biocompatibility studies showed that coated polyglactin 910 suture with triclosan was non-cytotoxic, non-irritating, and not a chemical pyrogen. In addition, an intramuscular implantation study demonstrated a tissue reaction, a healing response, and an absorption profile comparable to current polyglactin 910 suture. CONCLUSION: The extensive toxicology database supporting the safety of triclosan and the biocompatibility studies conducted on coated polyglactin 910 suture with triclosan demonstrate the safety of this suture for clinical use. Considering the clinical relevance of surgical site infections and the relatively low level of triclosan required to inhibit bacterial colonization of the suture, the use of this antimicrobial technology is well suited to this application. PMID- 12573041 TI - Physical and functional comparison of Coated VICRYL* Plus Antibacterial Suture (coated polyglactin 910 suture with triclosan) with Coated VICRYL* Suture (coated polyglactin 910 suture). AB - BACKGROUND: The goal of this study was to compare the physical and functional properties of coated polyglactin 910 suture with and without triclosan by human assessment and instrument-based measurements. METHODS: Surgeons specializing in general, orthopedic, plastic, or gynecologic surgery evaluated the suture materials in an in vivo porcine model with regard to (1) ease of passage through tissue, (2) first-throw knot holding, (3) knot tie-down smoothness, (4) knot security, (5) surgical handling, and (6) overall evaluation. Breaking strength retention was determined at 14, 21, 28, and 35 days post-implantation in rats using a tensile strength measurement device. The absorption rate was determined in rats by histopathology at 7, 28, 56, 63, 70, and 77 days post-implantation. The tactile smoothness and tie-down behavior of both wet and dry sutures were evaluated by product characterization technicians. RESULTS: The scores for surgeons' evaluation of suture material were favorable and similar for both sutures. Surgeons could not reliably make a distinction in handling between the two sutures. Breaking strength retention was the same for both sutures, ranging from 79% on day 14 to 5% on day 35. Both sutures were essentially absorbed at 70 days post-implantation. Product characterization assessment of the two sutures found them to be indistinguishable. CONCLUSION: The addition of triclosan to coated polyglactin 910 sutures did not affect physical handling properties or performance characteristics based on the testing and evaluations performed. PMID- 12573042 TI - In vitro antimicrobial evaluation of Coated VICRYL* Plus Antibacterial Suture (coated polyglactin 910 with triclosan) using zone of inhibition assays. AB - BACKGROUND: In this study, coated polyglactin 910 suture with triclosan was evaluated for its ability to inhibit the growth of wild-type and methicillin resistant Staphylococcus aureus and S. epidermidis using several in vitro models. METHODS: In the standard experimental procedure, sterile sutures were inoculated with bacteria, plated in semisolid culture media, incubated for 24 h, and examined for zones of inhibition surrounding the sutures. Sutures were pretreated by several methods prior to inoculation, these included: 24-h aqueous immersion, serial 7-day aqueous immersion, knotting, and passage through subcutaneous and fascial tissue. Sutures were also compared to determine if suture diameter had an effect on the zone of inhibition. RESULTS: Coated polyglactin 910 suture with triclosan inhibited growth of test organisms consistently over a range of suture diameters and treatment conditions. The antibacterial effect was robust and did not diminish when sutures were placed in an aqueous environment for up to 7 days. Knotted suture with triclosan inhibited bacterial colonization after replicate inoculation. The bacteria-free zone surrounding each knotted suture had a volume of 14.5 cm(3) for S. epidermidis and 17.8 cm(3) for S. aureus. Coated polyglactin 910 suture with triclosan continued to produce zones of inhibition after five and 10 passes through fascia and subcutaneous tissue. CONCLUSION: These data support the conclusion that coated polyglactin 910 suture with triclosan provides antimicrobial effect sufficient to prevent in vitro colonization by S. aureus and S. epidermidis. PMID- 12573043 TI - A 28-day study of the effect of Coated VICRYL* Plus Antibacterial Suture (coated polyglactin 910 suture with triclosan) on wound healing in guinea pig linear incisional skin wounds. AB - BACKGROUND: This study evaluated the effect of coated polyglactin 910 suture with triclosan on tissue response and wound healing over a 28-day period on full thickness linear wounds in the hairless guinea pig model. METHODS: In situ wound strength was assessed, and histopathologic effects on tissue response and wound healing were evaluated. Four (1.9 cm) full-thickness incisional wounds were created on the dorsolateral region (two left and two right, approximately 3 cm apart) in 40 Hartley male hairless guinea pigs (400-500 g). A 1.9-cm segment of suture material was implanted "length-wise" into the incision and the sites closed with simple interrupted closure with 3-0 nylon sutures. The test material was 2-0 coated polyglactin 910 suture with triclosan and the control material was 2-0 Coated VICRYL* Suture (coated polyglactin 910 suture). On days 3, 7, 14, and 28 postimplantation (n = 10/time interval), body weights were recorded, tissue specimens harvested, and bursting strength testing performed. RESULTS: There was no significant difference (P > 0.05) in bursting strength between the study groups for any of the time intervals studied. The bursting strengths (mm Hg) for the sutures with triclosan were 95.8 (day 3), 268.8 (day 7), 542.6 (day 14), and 633.8 (day 28). Both materials demonstrated comparable tissue response to implantation, and there were no significant differences (P > 0.05) observed in wound healing based on cellular response and collagen formation and orientation. CONCLUSION: Based on the in vivo biomechanical testing and histological results, no evidence of impedance to wound healing was detected. PMID- 12573044 TI - Panel discussion: current issues in the prevention and management of surgical site infection--part 2. PMID- 12573046 TI - Liposomal delivery of nucleic acids in vivo. AB - Optimization of cationic liposomal complexes for in vivo applications and therapeutics is complex involving many distinct components. These components include nucleic acid purification, plasmid design, formulation of the delivery vehicle, administration route and schedule, dosing, detection of gene expression, and others. This review will focus on optimization of these components for use in a variety of in vivo applications. Use of improved liposome formulations for delivery in vivo is valuable for gene therapy and would avoid several problems associated with viral delivery. Delivery of nucleic acids using liposomes is promising as a safe and non-immunogenic approach to gene therapy. Furthermore, gene therapeutics composed of artificial reagents can be standardized and regulated as drugs rather than as biologics. Optimizing all components of the delivery system will allow broad use of liposomal complexes to treat or cure human diseases or disorders. PMID- 12573047 TI - In vivo DNA electrotransfer. AB - The use of electrotransfer for DNA delivery to prokaryotic cells, and eukaryotic cells in vitro, has been well known and widely used for many years. However, it is only recently that electric fields have been used to enhance DNA transfer to animal cells in vivo, and this is known as DNA electrotransfer or in vivo DNA electroporation. Some of the advantages of this method of somatic cell gene transfer are that it is a simple method that can be used to transfer almost any DNA construct to animal cells and tissues in vivo; multiple constructs can be co transfected; it is equally applicable to dividing and nondividing cells; the DNA of interest does not need to be subcloned into a specific viral transfer vector and there is no need for the production of high titre viral stocks; and, as no viral genes are expressed there is less chance of an adverse immunologic reaction to vector sequences. The ease with which efficient in vivo gene transfer can be achieved with in vivo DNA electrotransfer is now allowing genetic analysis to be applied to a number of classic animal model systems where transgenic and embryonic stem cell techniques are not well developed, but for which a wealth of detailed descriptive embryological information is available, or surgical manipulation is much more feasible. As well as exciting applications in developmental biology, in vivo DNA electrotransfer is also being used to transfer genes to skeletal muscle and drive expression of therapeutically active proteins, and to examine exogenous gene and protein function in normal adult cells situated within the complex environment of a tissue and organ system in vivo. Thus, in effect providing the in vivo equivalent of the in vitro transient transfection assay. As the widespread use of in vivo electroporation has really only just begun, it is likely that the future will hold many more applications for this technology in basic research, biotechnology and clinical research areas. PMID- 12573048 TI - Taking the cell by stealth or storm? Protein transduction domains (PTDs) as versatile vectors for delivery. AB - A cell delivery system is increasing in use in many areas of cell and molecular biology and bio-medicine. This system is based on a number of naturally occurring protein motifs and/or sequences which show the remarkable ability to rapidly cross the mammalian cell membrane without compromising its structure or function. These so-called Protein Transduction Domains (PTDs) offer unprecedented advantages for intracellular delivery. These advantages include, but are not limited to, their applicability to all cell types (no cell type has yet been described which is not transduced by these PTDs), and the range of cargoes that can be transduced (including peptides, small proteins, full-length enzymes, DNA oligomers, peptide-nucleic acid oligomers, liposomes, and magnetic nanoparticles). Furthermore, the PTDs have been demonstrated to be suitable for in vivo delivery including delivery across the blood brain barrier, and have been shown to cross the plasma membrane rapidly and enter the cytoplasm and nuclear regions of the cell. In this review, the general properties of the most commonly used PTDs are described. The strategies currently being undertaken also highlight that improvements in membrane transduction are possible despite our lack of understanding of the exact biochemical and/or physical mechanisms of transduction. Recent examples of the range of potential applications are also discussed. PMID- 12573049 TI - Adenovirus and adeno-associated virus vectors. AB - Recombinant adenovirus (rAd) and recombinant adeno-associated virus (rAAV) are among the most extensively used vectors in gene therapy studies to date. These two vectors share some similar features such as a broad host range and ability to infect both proliferating and quiescent cells. However, they also possess their own unique set of properties that render them particularly attractive for gene therapy applications. rAd vectors can accommodate larger inserts, mediate transient but high levels of protein expression, and can be easily produced at high titers. Development of gutted rAd vectors has further increased the cloning capacity of these vectors. The gaining popularity of rAAV use in gene therapy can be attributed to its lack of pathogenicity and added safety due to its replication defectiveness, and its ability to mediate long-term expression in a variety of tissues. Site-specific integration, as occurs with wild-type AAV, will be a unique and valuable feature if incorporated into rAAV vectors, further improving their safety. This paper describes these properties of rAd and rAAV vectors, and discusses further development and vector improvements that continue to extend the utility of these vectors, such as cell retargeting by capsid modification, differential transduction by use of serotypes, and extension of the cloning capacity of rAAV vectors by dual vector heterodimerization. PMID- 12573050 TI - Gene delivery using herpes simplex virus vectors. AB - Herpes simplex virus (HSV) is a neurotropic DNA virus with many favorable properties as a gene delivery vector. HSV is highly infectious, so HSV vectors are efficient vehicles for the delivery of exogenous genetic material to cells. Viral replication is readily disrupted by null mutations in immediate early genes that in vitro can be complemented in trans, enabling straightforward production of high-titre pure preparations of non-pathogenic vector. The genome is large (152 Kb) and many of the viral genes are dispensable for replication in vitro, allowing their replacement with large or multiple transgenes. Latent infection with wild-type virus results in episomal viral persistence in sensory neuronal nuclei for the duration of the host lifetime. Transduction with replication defective vectors causes a latent-like infection in both neural and non-neural tissue; the vectors are non-pathogenic, unable to reactivate and persist long term. The latency active promoter complex can be exploited in vector design to achieve long-term stable transgene expression in the nervous system. HSV vectors transduce a broad range of tissues because of the wide expression pattern of the cellular receptors recognized by the virus. Increasing understanding of the processes involved in cellular entry has allowed preliminary steps to be taken towards targeting the tropism of HSV vectors. Using replication-defective HSV vectors, highly encouraging results have emerged from recent pre-clinical studies on models of neurological disease, including glioma, peripheral neuropathy, chronic pain and neurodegeneration. Consequently, HSV vectors encoding appropriate transgenes to tackle these pathogenic processes are poised to enter clinical trials. PMID- 12573051 TI - Lentiviral vectors for gene delivery into cells. AB - Human immunodeficiency virus type I (HIV) is the etiologic agent of acquired immunodeficiency syndrome or AIDS. Vectors based upon HIV have been in use for over a decade. Beginning in 1996, with the demonstration of improved pseudotyping using vesicular stomatitis virus (VSV) G protein along with transduction of resting mammalian cells, a series of improvements have been made in these vectors, making them both safer and more efficacious. Taking a cue from vector development of murine leukemia virus (MLV), split coding and self-inactivating HIV vectors now appear quite suitable for phase I clinical trials. In parallel, a number of pre-clinical efficacy studies in animals have demonstrated the utility of these vectors for various diseases processes, especially neurodegenerative and hematopoietic illnesses. These vectors are also appropriate for the study of other viruses (specifically of viral entry) and investigation of the HIV replicative cycle, along with straightforward transgene delivery to target cells of interest. Vectors based upon other lentiviruses have shown similar abilities and promise. Although concerns remain, particularly with regards to detection and propagation of replication-competent lentivirus, it is almost certain that these vectors will be introduced into the clinic within the next 3-5 years. PMID- 12573053 TI - Microparticle bombardment as a tool in plant science and agricultural biotechnology. AB - Microparticle bombardment technology has evolved as a method for delivering exogenous nucleic acids into plant cells and is a commonly employed technique in plant science. Desired genetic material is precipitated onto micron-sized metal particles and placed within one of a variety of devices designed to accelerate these "microcarriers" to velocities required to penetrate the plant cell wall. In this manner, transgenes can be delivered into the cell's genome or plastome. Since the late 1980s microparticle bombardment has become a powerful tool for the study of gene expression and production of stably transformed tissues and whole transgenic plants for experimental purposes and agricultural applications. This paper reviews development and application of the technology, including the protocols and mechanical systems employed as delivery systems, and the types of plant cells and culture systems employed to generate effective "targets" for receiving the incoming genetic material. Current understanding of how the exogenous DNA becomes integrated into the plant's native genetic background are assessed as are methods for improving the efficiency of this process. Pros and cons of particle bombardment technologies compared to alternative direct gene transfer methods and Agrobacterium based transformation systems are discussed. PMID- 12573052 TI - RNA based vaccines. AB - The recognition that CD8(+) T-cell mediated Th1 immune responses were necessary to produce immunity to intracellular and transformed self pathogens led to intense interest in the delivery of nucleic acids, DNA, or RNA encoding candidate antigens, as vaccines. Antigen presenting cells (APC) encounter most protein and vaccine immunogens as extracellular proteins and, thus, present them on major histocompatibility complex (MHC) class II molecules leading to the activation of CD4(+) T cells. Protein antigens encoded by nucleic acids delivered to dendritic cell (DC) are produced inside the cell and, thus, can stimulate MHC class I mediated activation of CD8(+) T-cell immune responses. Unfortunately, DCs are not readily transfected with DNA (Akbari et al., 1999) resulting in the requirement for high concentrations of DNA and repeated immunizations to achieved immune responses. RNA, on the other hand, is readily taken up and expressed by DC, making it an alternative vaccine candidate. In this article, we will discuss immune responses developed, interactions between APC and RNA that activate and dictate DC activation, and preliminary studies using RNA in vivo and in vitro to develop protective immunity. PMID- 12573054 TI - Intravenous nonviral gene therapy causes normalization of striatal tyrosine hydroxylase and reversal of motor impairment in experimental parkinsonism. AB - Brain gene-targeting technology is used to reversibly normalize tyrosine hydroxylase (TH) activity in the striatum of adult rats, using the experimental 6 hydroxydopamine model of Parkinson's disease. The TH expression plasmid is encapsulated inside an 85-nm PEGylated immunoliposome (PIL) that is targeted with either the OX26 murine monoclonal antibody (MAb) to the rat transferrin receptor (TfR) or with the mouse IgG2a isotype control antibody. TfRMAb-PIL, or mIgG2a PIL, is injected intravenously at a dose of 10 microg of plasmid DNA per rat. TfRMAb-PIL, but not mIgG2a-PIL, enters the brain via the transvascular route. The targeting TfRMAb enables the nanocontainer carrying the gene to undergo both receptor-mediated transcytosis across the blood-brain barrier (BBB) and receptor mediated endocytosis into neurons behind the BBB by accessing the TfR. With this approach, the striatal TH activity ipsilateral to the intracerebral injection of the neurotoxin was normalized and increased from 738 +/- 179 to 5486 +/- 899 pmol/hr per milligram of protein. The TH enzyme activity measurements were corroborated by TH immunocytochemistry, which showed that the entire striatum was immunoreactive for TH after intravenous gene therapy. The normalization of striatal biochemistry was associated with a reversal of apomorphine-induced rotation behavior. Lesioned animals treated with the apomorphine exhibited 20 +/- 5 and 6 +/- 2 rotations/min, respectively, after intravenous administration of the TH plasmid encapsulated in mIgG2a-PIL and TfRMAb-PIL. These studies demonstrate that it is possible to normalize brain enzyme activity by intravenous administration and nonviral gene transfer. PMID- 12573055 TI - Dendritic cells transduced to express interleukin-4 prevent diabetes in nonobese diabetic mice with advanced insulitis. AB - Our previous studies demonstrated that adoptive transfer of dendritic cells (DC) prevents diabetes in young nonobese diabetic (NOD) mice by inducing regulatory T(H)2 cells. In this report, as a means of treating NOD mice with more advanced insulitis, we infected DC with adenoviral vectors expressing interleukin (IL)-4 (Ad.IL-4), eGFP (Ad.eGFP), or empty vector (Ad psi 5). DC infected with any of the Ad vectors expressed higher levels of CD40, CD80, and CD86 molecules than uninfected DC and Ad.IL-4 DC produced IL-4 after lipopolysaccharide (LPS) and interferon (IFN)-gamma stimulation. Ad-infected DC efficiently stimulated allogeneic T cells, and cultures of T cells with Ad.IL-4 DC produced lower levels of IFN-gamma and marginally higher levels of IL-4. In vivo studies demonstrated that the Ad.eGFP DC trafficked to the pancreatic lymph nodes within 24 hr of intravenous administration, and could be visualized in the T cell areas of the spleen. The intrapancreatic IFN-gamma:IL-4 or IFN-gamma:IL-10 cytokine ratios were lower in 10-week-old mice treated with Ad.IL-4 DC, and these mice were significantly protected from disease. These results demonstrate, for the first time, that genetically modified DC can prevent diabetes in the context of advanced insulitis. PMID- 12573056 TI - Development of an adenovirus-shedding assay for the detection of adenoviral vector-based vaccine and gene therapy products in clinical specimens. AB - Adenoviral vectors are used widely as gene therapy and vaccine delivery systems. An adenovirus-shedding assay may be performed in clinical trials to monitor the safety of the vector and to investigate the potential relation between clinical symptoms and shed vector virus. This report describes the development and statistical performance of the shedding assay. Live adenovirus was recovered from throat swab and urine samples spiked with E1-deleted adenovirus type 5 vector expressing HIV-1 gag [Ad5HIVgag], in the presence or absence of wild-type adenovirus (WT Ad5). Samples were cultured in 293 and A549 cells, and the DNA extracted from virus culture was tested by polymerase chain reaction (PCR) for sequence identity. The results showed that the frequency of Ad5HIVgag infectivity in 293 cells by cytopathic effect (CPE) or an immunofluorescence assay (IFA) was concentration-dependent (53% for 10(2), 94% for 10(4), and 100% for 10(6) viral particles). WT Ad5 virus did not interfere with Ad5HIVgag. PCR amplisets could specifically amplify target sequences in the background of nonspecific DNA matrices and could distinguish Ad5HIVgag from wild-type adenoviruses. This assay may be used for clinical trials using adenovirus vectors as vehicles for vaccines. PMID- 12573057 TI - Efficient trans-splicing in the retina expands the utility of adeno-associated virus as a vector for gene therapy. AB - Recombinant vectors based on adeno-associated virus (AAV) can efficiently transduce many different cell types, including cells of the retina, resulting in stable gene expression. A major shortcoming of this vector is its small packaging capacity. A trans-splicing approach, which reconstitutes gene expression from two independent AAV vectors, can be used to overcome the vector's packaging limitations. The efficiency of this system to date has been disappointing, and therefore its utility for therapeutic application limited. We demonstrate here that efficiency and cellular specificity of trans-splicing is dependent on selection of the appropriate AAV serotype. Efficiency of transgene expression resulting from trans-splicing in skeletal muscle approaches that obtained when delivering the intact transgene when using AAV2 vectors packaged with AAV5 capsids (AAV2/5). This expands the potential of AAV vectors for retinal gene therapy. The use of AAV2/5 also increases the efficiency of trans-splicing in photoreceptors. Selection of the appropriate AAV serotype is likely to affect efficiency of trans-splicing in other organ systems as well. PMID- 12573058 TI - Treatment of human papillomavirus (HPV) type 16-infected cells using herpes simplex virus type 1 thymidine kinase-mediated gene therapy transcriptionally regulated by the HPV E2 protein. AB - Human papillomavirus type 16 (HPV-16) is associated with development of anogenital squamous cell cancers (SCCs) and their precursor, intraepithelial neoplasia (IN). Few approaches to the treatment of IN to prevent SCC are targeted specifically to HPV. We have designed an HPV-specific therapy using the herpes simplex virus type 1 thymidine kinase (HSV-1 TK) gene driven by an HPV-specific promoter in the HPV-16 long control region (LCR) (nucleotide 7450-nucleotide 104), which is regulated by the HPV E2 protein. Expression of the HSV-1 TK gene is designed to render HPV-infected cells sensitive to the prodrugs ganciclovir (GCV) and acyclovir (ACV). To assess the E2 specificity of gene expression driven by the HPV-16 LCR, we measured luciferase expression in HPV-positive and HPV negative cell lines. Significant induction of luciferase activity was observed in HPV-positive cells when compared with four different HPV-negative epithelial cell lines. Cotransfection of an HPV-negative cell line, MDCK, with an HPV-16 E2 expressing plasmid resulted in 15- to 20-fold induction of luciferase activity, suggesting specific activation by E2 protein. A plasmid expressing the HSV-1 TK gene driven by the LCR was transfected into CaSki and SiHa cells. Treatment of transfected cells with either GCV or ACV (20-30 microg/ml) for 6-10 days resulted in 80-95% cell death. Cell death was progressive, dose dependent, and mediated by apoptosis. These results suggest that direct gene transfer of the HSV-1 TK gene into HPV-16-infected cells expressing E2 protein, accompanied by treatment with either GCV or ACV, may be a clinically feasible therapeutic strategy. PMID- 12573059 TI - Multilineage cells from adipose tissue as gene delivery vehicles. AB - We have characterized a population of mesenchymal progenitor cells from adipose tissue, termed processed lipoaspirate (PLA) cells, which have multilineage potential similar to bone marrow-derived mesenchymal stem cells and are also easily expanded in culture. The primary benefit of using adipose tissue as a source of multilineage progenitor cells is its relative abundance and ease of procurement. We examined the infection of PLA cells with adenoviral, oncoretroviral, and lentiviral vectors. We demonstrate that PLA cells can be transduced with lentiviral vectors at high efficiency. PLA cells maintain transgene expression after differentiation into adipogenic and osteogenic lineages after lentiviral transduction. Therefore, PLA cells and lentiviral vectors may be an efficient combination for use as a therapeutic gene delivery vehicle. PMID- 12573060 TI - Large-scale production of pseudotyped lentiviral vectors using baculovirus GP64. AB - Unlike oncoretroviruses, lentiviral vectors can insert large genes and can target both dividing and nondividing cells; thus they hold unique promise as gene transfer agents. To enhance target range, the native lentiviral envelope glycoprotein is replaced (pseudotyped) with vesicular stomatitis virus G (VSVG), and the genes of interest are packaged in nonreplicating vectors by transient transfection with three plasmids. However, because of cytotoxic effects of VSVG expression in producer cells (293T cells) it has been difficult to generate a packaging cell line, required for even modest scale-up of vector production. Here we introduce a pseudotyped lentivirus vector using the baculovirus GP64 envelope glycoprotein. Compared with VSVG, GP64 vectors exhibited a similar broad tropism and similar native titers. GP64-pseudotyped vectors were usually highly concentrated without much loss of titer. Because, unlike VSVG, GP64 expression does not kill cells, we generated 293T-based cell lines constitutively expressing GP64. Our results demonstrate that the baculovirus GP64 protein is an attractive alternative to VSVG for viral vectors used in the large-scale production of high titer virus required for clinical and commercial applications. PMID- 12573061 TI - Gene transfer to subdermal tissues via a new gene gun design. AB - Although particle-mediated gene transfer technology (gene gun) has been applied for gene transfer to external tissues, the application of this technology to other tissues has met with limited success. Here we report the development of a new design of a gene gun that uses helium discharge to propel DNA-coated gold beads that are suspended in liquid. Higher discharge pressures allow for the delivery of DNA to deeper tissues. Using the new gene gun to deliver a luciferase expression plasmid resulted in higher levels of gene expression in the skin than observed with conventional guns, as well as in subdermal tissues, including subcutaneous tumors. Even when using as little as 125 ng of DNA, gene expression in skin and muscle reached its peak level at 24 hr postbombardment and remained for at least 1 week. The use of a LacZ expression plasmid showed that gene expression was distributed throughout the skin with no observable pathology. The new gene gun was used to deliver a model tumor rejection antigen (a modified human papilloma virus [HPV] E7 gene) to mice. All of the treated animals developed protective immunity against HPV-positive tumors. These results demonstrate that our new design can be used in standard gene gun applications and extends the reach of gene gun technology to tissues that were previously unavailable. PMID- 12573062 TI - Phylogeography of Lake Malawi cichlids of the genus Pseudotropheus: significance of allopatric colour variation. AB - One of the most compelling features of the cichlid fishes of the African Great Lakes is the seemingly endless diversity of male coloration. Colour diversification has been implicated as an important factor driving cichlid speciation. Colour has also been central to cichlid taxonomy and, thus, to our concept of species diversity. We undertook a phylogeographical examination of several allopatric populations of the Lake Malawi cichlid Pseudotropheus zebra in order to reconstruct the evolutionary history of the populations, which exhibit one of two dorsal fin colours. We present evidence that populations with red dorsal fins (RT) are not monophyletic. The RT population defining the northern limit of the distribution has evidently originated independently of the southern RT populations, which share a common ancestry. This evidence of species-level colour convergence is an important discovery in our understanding of cichlid evolution. It implies that divergence in coloration may accompany speciation, and that allopatric populations with similar coloration cannot be assumed to be conspecific. In addition to this finding, we have observed evidence for introgression, contributing to current evidence that this phenomenon may be extremely widespread. Thus, in species-level phylogenetic reconstructions, including our own, consideration must be given to the potential effects of introgression. PMID- 12573063 TI - Active formation of mixed-species grouse leks: a role for predation in lek evolution? AB - Behavioural ecologists have interpreted avian leks as products of sexual selection, in which males display socially to increase their opportunities to mate. However, without invoking reproductive queuing or kin selection, this paradigm does not necessarily explain why many males that fail to mate participate in leks. An alternative solution, that males also aggregate to reduce predation, has previously lacked compelling support. We show that mixed-species leks, comprising two congeneric grouse, form when single males or small groups of one species, the greater prairie chicken Tympanuchus cupido, join leks of another, the sharp-tailed grouse T. phasianellus. We documented the process by observing lek dynamics and comparing group sizes between mixed- and single species leks. Joining implies that prairie chickens benefit from displaying with sharp-tailed grouse. The numbers of females of each species attending a lek increased with the number of conspecific, but not heterospecific, males. This suggests that the joining of heterospecifics is unlikely to increase mating opportunities, and leaves lowered predation risk as the most likely benefit of associating with heterospecifics. Active formation of mixed-species leks therefore suggests that predation may be sufficient to drive lek formation. The benefits of participation in mixed leks may be asymmetrical because prairie chickens display more and are less vigilant than sharp-tailed grouse. PMID- 12573064 TI - Space and the persistence of male-killing endosymbionts in insect populations. AB - Male-killing bacteria are bacteria that are transmitted vertically through the females of their insect hosts. They can distort the sex ratio of their hosts by killing infected male offspring. In nature, male-killing endosymbionts (male killers) often have a 100% efficient vertical transmission, and multiple male killing bacteria infecting a single population are observed. We use different model formalisms to study these observations. In mean-field models a male killer with perfect transmission drives the host population to extinction, and coexistence between multiple male killers within one population is impossible; however, in spatially explicit models, both phenomena are readily observed. We show how the spatial pattern formation underlies these results. In the case of high transmission efficiencies, waves with a high density of male killers alternate with waves of mainly wild-type hosts. The male killers cause local extinction, but this creates an opportunity for uninfected hosts to re-invade these areas. Spatial pattern formation also creates an opportunity for two male killers to coexist within one population: different strains create spatial regions that are qualitatively different; these areas then serve as different niches, making coexistence possible. PMID- 12573065 TI - Song and the song control pathway in the brain can develop independently of exposure to song in the sedge warbler. AB - Previous studies have shown that female sedge warblers choose to mate with males that have more complex songs, and sexual selection has driven the evolution of both song complexity and the size of the major song control area (HVc) in the brain. In songbirds, learning from conspecifics plays a major role in song development and this study investigates the effects of isolation and exposure to song on song structure and the underlying song control system. Sibling pairs of hand-reared nestling sedge warblers were reared to sexual maturity under two conditions. Siblings in one group were reared individually in acoustic isolation in separate soundproof chambers. In the other group, siblings were reared together in an aviary with playback of recorded songs. The following spring, analysis of songs revealed that siblings reared in acoustic isolation produced normal song structures, including larger syllable repertoires than those exposed to song. We found no significant differences in the volumes of HVc, nucleus robustus archistnatalis, the lateral portion of the magnocellular nucleus and the density of dendritic spines between the two groups. Males exceeded females in all these measures, and also had a larger telencephalon. Our experiments show that complex song, sexual dimorphism in brain structure, and the size of song nuclei can all develop independently of exposure to song. These findings have important implications for how sexual selection can operate upon a complex male trait such as song and how it may also shape the more general evolution of brain structure in songbirds. PMID- 12573066 TI - Directional female preference for an exaggerated male trait in canary (Serinus canaria) song. AB - Motor constraints on vocal production impose a trade-off between trill rate and frequency bandwidth within birdsong. We tested whether domesticated canary (Serinus canaria) females, reared either in acoustic isolation or in aviary conditions, had a preference for broad bandwidth songs with artificially increased syllable rates. The copulation solicitation display (CSD) was used as an index of female preference. As predicted, both naive and experienced females were especially responsive to syllables with a broad bandwidth emitted at an artificially increased rate. Female preference for supernormal stimuli provide support for the honest-signalling hypothesis and our results are consistent with recent findings indicating that production of song phrases maximizing both bandwidth and syllable rate may be a reliable indicator of male physical or behavioural qualities. We suggest that female preference for vocal emissions, which simultaneously maximize these two parameters, could be a widespread pattern within songbirds. PMID- 12573067 TI - Validation of Bateman's principles: a genetic study of sexual selection and mating patterns in the rough-skinned newt. AB - Few studies have influenced thought on the nature of sexual selection to the extent of the classic paper of A. J. Bateman on mating patterns in Drosophila. However, interpretation of his study remains controversial, and a lack of modern empirical evidence prevents a consensus with respect to the perceived utility of Bateman's principles in the study of sexual selection. Here, we use a genetic study of natural mating patterns in the rough-skinned newt, Taricha granulosa, to investigate the concordance between Bateman's principles and the intensity of sexual selection. We found that males experienced strong sexual selection on tail height and body size, while sexual selection was undetectable in females. This direct quantification of sexual selection agreed perfectly with inferences that are based on Bateman's principles. Specifically, males (in comparison with females) exhibited greater standardized variances in reproductive and mating success, as well as a stronger relationship between mating success and reproductive success. Overall, our results illustrate that Bateman's principles provide the only quantitative measures of the mating system with explicit connections to formal selection theory and should be the central focus of studies of mating patterns in natural populations. PMID- 12573068 TI - Interference competition and species coexistence. AB - Interference competition is ubiquitous in nature. Yet its effects on resource exploitation remain largely unexplored for species that compete for dynamic resources. Here, I present a model of exploitative and interference competition with explicit resource dynamics. The model incorporates both biotic and abiotic resources. It considers interference competition both in the classical sense (i.e. each species suffers a net reduction in per capita growth rate via interference from, and interference on, the other species) and in the broad sense (i.e. each species suffers a net reduction in per capita growth rate via interference from, but can experience an increase in growth rate via interference on, the other species). Coexistence cannot occur under classical interference competition even when the species inferior at resource exploitation is superior at interference. Such a trade-off can, however, change the mechanism of competitive exclusion from dominance by the superior resource exploiter to a priority effect. Now the inferior resource exploiter can exclude the superior resource exploiter provided it has a higher initial abundance. By contrast, when interference is beneficial to the interacting species, coexistence is possible via a trade-off between exploitation and interference. These results hold regardless of whether the resource is biotic or abiotic, indicating that the outcome of exploitative and interference competition does not depend on the exact nature of resource dynamics. The model makes two key predictions. First, species that engage in costly interference mechanisms (e.g. territoriality, overgrowth or undercutting, allelopathy and other forms of chemical competition) should not be able to coexist unless they also engage in beneficial interference mechanisms (e.g. predation or parasitism). Second, exotic invasive species that displace native biota should be superior resource exploiters that have strong interference effects on native species with little or negative cost. The first prediction provides a potential explanation for patterns observed in several natural systems, including plants, aquatic invertebrates and insects. The second prediction is supported by data on invasive plants and vertebrates. PMID- 12573069 TI - Interspecific competition during transmission of two sympatric malaria parasite species to the mosquito vector. AB - The role of species interactions in structuring parasite communities remains controversial. Here, we show that interspecific competition between two avian malaria parasite species, Plasmodium gallinaceum and P. juxtanucleare, occurs as a result of interference during parasite fertilization within the bloodmeal of the mosquito. The significant reduction in the transmission success of P. gallinaceum to mosquitoes, due to the co-infecting P. juxtanucleare, is predicted to have compromised its colonization of regions occupied by P. juxtanucleare and, thus, may have contributed to the restricted global distribution of P. gallinaceum. Such interspecies interactions may occur between human malaria parasites and, thus, impact upon parasite species epidemiology, especially in regions of seasonal transmission. PMID- 12573070 TI - Kin competition within groups: the offspring depreciation hypothesis. AB - Where relatives compete for the same resources (kin competition) and each obtains an equal share, this can favour the evolution of elevated dispersal rates, such that most resource competition is among non-relatives. We show that this effect evaporates as among-sibling dominance increases to the point where the allocation of resources is maximally unequal. We restore a kin-competition effect on emigration rates from dominance-ranked family groups by showing that where siblings form queues to inherit the breeding positions, the length of the queue affects the fitness of all individuals by depreciating the rank of subsequent offspring. Incorporating this 'offspring depreciation' effect decreases optimal queue lengths, increases dispersal rates and explains the otherwise paradoxical use of sinks by cooperatively breeding birds in stable environments. The offspring depreciation effect also favours the evolution of small, but consistent, clutch sizes and high reproductive skew, but constrains the evolution of alloparenting. PMID- 12573071 TI - Pollen competition as a unilateral reproductive barrier between sympatric diploid and tetraploid Chamerion angustifolium. AB - Speciation requires the evolution of barriers to gene exchange between descendant and progenitor populations. Cryptic reproductive barriers in plants arise after pollination but before fertilization as a result of pollen competition and interactions between male gametophytes and female reproductive tissues. We tested for such gametic isolation between the polyploid Chamerion angustifolium and its diploid progenitor by conducting single (diploid or tetraploid) and mixed ploidy (1 : 1 diploid and tetraploid) pollinations on both cytotypes and inferring siring success from paternity analysis and pollen-tube counts. In mixed pollinations, polyploids sired most (79%) of their own seeds as well as those of diploids (61%) (correcting for triploid block, siring success was 70% and 83%, respectively). In single donor pollinations, pollen tubes from tetraploids were more numerous than those from diploids at four different positions in each style and for both diploid and tetraploid pollen recipients. The lack of a pollen donor x recipient interaction indicates that the tetraploid siring advantage is a result of pollen competition rather than pollen-pistil interactions. Such unilateral pollen precedence results in an asymmetrical pattern of isolation, with tetraploids experiencing less gene flow than diploids. It also enhances tetraploid establishment in sympatric populations, by maximizing tetraploid success and simultaneously diminishing that of diploids through the production of inviable triploid offspring. PMID- 12573072 TI - Pattern discrimination in a hawkmoth: innate preferences, learning performance and ecology. AB - Spatial patterns are important cues for flower detection and recognition by nectar-feeding insects. Pattern vision has been studied in much detail in bees and flies but rarely in butterflies and moths. In this paper, I present a first proof of pattern-learning abilities in a moth, and discuss reasons for the limitations to their pattern learning. The diurnal hawkmoth Macroglossum stellatarum spontaneously prefers patterned to uniformly coloured stimuli but can be trained to choose the uniform stimulus. By contrast, experience does not override the innate preferences for radial over tangential patterns, and for tangential over striped patterns. These results do not reflect bad visual discrimination but rather a lack of learning ability and motivation to abolish innate preferences. I propose that radial and tangential flower patterns are good predictors of nectar reward, a condition under which learning is unlikely to evolve. These patterns serve not only as cues for flower detection but also as guides to the reward. Hovering pollinators strongly depend on these guides and should therefore: (i). have rigid pattern preferences; and (ii). not be motivated to abolish these preferences as easily as their innate preferences for colours. PMID- 12573073 TI - Population dynamics of a South American rodent: seasonal structure interacting with climate, density dependence and predator effects. AB - Understanding the role of interactions between intrinsic feedback loops and external climatic forces is one of the central challenges within the field of population ecology. For rodent dynamics, the seasonal structure of the environment necessitates changes between two stages: reproductive and non reproductive. Nevertheless, the interactions between seasonality, climate, density dependence and predators have been generally ignored. We demonstrate that direct climate effects, the nonlinear effect of predators and the nonlinear first order feedback embedded in a seasonal structure are key elements underlying the large and irregular fluctuations in population numbers exhibited by a small rodent in a semi-arid region of central Chile. We found that factors influencing population growth rates clearly differ between breeding and non-breeding seasons. In addition, we detected nonlinear density dependencies as well as nonlinear and differential effects of generalist and specialist predators. Recent climatic changes may account for dramatic perturbations of the rodent's population dynamics. Changes in the predator guild induced by climate are likely to result, through the food web, in a large impact on small rodent demography and population dynamics. Assuming such interactions to be typical of ecological systems, we conclude that appropriate predictions of the ecological consequences of climate change will depend on having an in-depth understanding of the community-weather system. PMID- 12573074 TI - Scaling in stream communities. AB - Scaling relationships between population density (N) and body size (W), and of their underlying size distributions, can contribute to an understanding of how species use resources as a function of size. In an attempt to resolve the controversy over the form of scaling relationships, an extensive dataset, comprising 602 invertebrate species, was obtained from two geographically separate stream communities (Seebach in Austria and Mynach in Wales). We analysed the temporal consistency of the N-W relationship, which was subjected to ordinary least squares (OLS), bisector (OLS(BIS)) and quantile regressions, and species size spectra with seasonally collated data. Slopes of seasonal OLS(BIS) regressions did not depart from -1 in either community, indicating a seasonally convergent scaling relationship, which is not energetically constrained. Species size spectra may scale with habitat complexity, providing an alternative explanation for the observed body-size scaling. In contrast to the right-skewed species-size frequency distributions of single-species assemblages, the size spectra of these benthic communities exhibited 'central tendencies', reflecting their phyletic constitution. The shape of species body-mass spectra differed between the two communities, with a bimodal and seasonally convergent pattern in the Seebach community and a seasonally shifting unimodality in the Mynach community. The body-size spectra of large, mostly insect, species (greater than or equal to 1 mm) scaled to seasonal variations in habitat complexity (i.e. fractal D), suggesting that habitat structure constrains the community organization of stream benthos. PMID- 12573075 TI - Negative feedback within a mutualism: host-specific growth of mycorrhizal fungi reduces plant benefit. AB - A basic tenet of ecology is that negative feedback on abundance plays an important part in the coexistence of species within guilds. Mutualistic interactions generate positive feedbacks on abundance and therefore are not thought to contribute to the maintenance of diversity. Here, I report evidence of negative feedback on plant growth through changes in the composition of their mutualistic fungal symbionts, arbuscular mycorrhizal (AM) fungi. Negative feedback results from asymmetries in the delivery of benefit between plant and AM fungal species in which the AM fungus that grows best with the plant Plantago lanceolata is a poor growth promoter for Plantago. Growth of Plantago is, instead, best promoted by the AM fungal species that accumulate with a second plant species, Panicum sphaerocarpon. The resulting community dynamic leads to a decline in mutualistic benefit received by Plantago, and can contribute to the coexistence of these two competing plant species. PMID- 12573076 TI - Sexual selection as a cause of human skin colour variation: Darwin's hypothesis revisited. AB - The dark skin of tropical peoples is likely to be an adaptation to the strong ultraviolet (UV) radiation near the equator, perhaps protecting against sunburn or degradation of folate. By contrast, the adaptive value of light skin is questionable. In particular, the relevance of vitamin D deficiency rickets as a selective factor has been cogently criticized. Population genetic studies on the melanocortin 1 receptor (MC1R) gene (one of the genes responsible for normal human skin colour variation) also cast doubt on the role of positive natural selection in the evolution of light skin. Natural selection may favour dark skin everywhere, though to a lesser extent at higher latitudes. Darwin believed that racial differences in skin colour were caused by sexual selection. Available evidence suggests that in each society a lighter-than-average skin colour is preferred in a sexual partner. Such a preference would generate sexual selection for light skin that counteracts natural selection for dark skin. The observed latitudinal gradient in skin colour may result from the balance between natural and sexual selection. PMID- 12573077 TI - Undernutrition prevalence and social determinants in children aged 0-59 months, Niteroi, Brazil. AB - OBJECTIVE: With the aim of diagnosing the nutritional status and nutritional social-economic determinants in children aged 0-59 months, 2194 children were studied in the three Health Districts of the city of Niteroi, Rio de Janeiro State, Brazil. MATERIALS AND METHODS: Data were obtained through a random sampling survey performed in the city's vaccination centres. Variables collected were maternal schooling, family income, dwelling's connection to safe water, dwelling's sewage connection, birthweight and the anthropometric indicators height/age (H/A; for stunting), weight/height (W/H; for wasting) and weight/age (W/A; for underweight). Undernutrition risk factors were identified with the help of logistic regression models for the age ranges 0-12 and 13-36 months. RESULTS: Stunting and underweight levels were found to be above those of the NCHS reference, although, in general, at levels better than those frequently found in Third World countries. Low birthweight prevalence in the city was 9%, also above common levels in more developed countries. Undernutrition risks were, in general, higher for the younger age range. The variables family income, maternal schooling, water connection and birthweight could be characterized as important undernutrition risk factors, being most noticeable the effects of family income and birthweight both for the '0-12 months' and for the 'above 13 months' age ranges. CONCLUSIONS: Although the city's problems are mostly concentrated on chronic undernutrition, more attention should be given to children's birthweight deficits, which appear to have effects on children's growth that extend for years after birth. PMID- 12573078 TI - The use of body mass index to predict body composition in children. AB - BACKGROUND: Simple anthropometric indices of body composition have particular appeal for use in children, and as such body mass index (BMI) has been used to predict percentage body fat in a number of studies. AIM: To evaluate the relationship between BMI and percentage body fat (%body fat) and a proposed, more appropriate relationship between BMI and fat mass/height(2) in a cohort of young children. SUBJECTS AND METHODS: Cross-sectional study of 109 children aged between 6 and 10 years residing in either Sydney or Brisbane, Australia. Weight and height were measured using standard methods. Body composition was measured using a stable isotope method to firstly determine total body water and subsequently fat free mass. RESULTS: The correlation between BMI and fat mass/height(2) was markedly greater than that between BMI and percentage body fat. In the entire group of children the R(2) (x100%) value for the relationship between BMI and fat mass/height(2) was 73.3% compared with 46.5% for the relationship between BMI and percentage body fat. CONCLUSIONS: We have shown that the use of BMI to predict fat mass/height(2), and consequently percentage body fat, is superior to the use of BMI to predict percentage body fat based directly upon the R(2) values of the above analysis. PMID- 12573079 TI - The relative strength of weight and length at birth in contrast to social factors as determinants of height at 18 years in Brazil. AB - BACKGROUND: There is scarce information on the relative importance of measurements at birth and social factors in determining adult height. AIM: To assess the relative contribution of length and weight at birth, and social factors to height at 18 years. METHODS: A total of 3470 boys born in Ribeirao Preto, Brazil, in 1978/1979 were reassessed at the time of military conscription. Anthropometric, demographic and social variables were collected concurrently and used in a multiple regression analysis. RESULTS: A total of 2029 (61.3%) of the target population were assessed at 18 years. In the adjusted analysis, birth length, birth weight, social class, neighbourhood wealth of subject, maternal age and birth order were highly associated with height at 18 years. The results were similar when birth weight and length at birth were included as continuous or categorical variables. The differences in height due to length at birth were greater than those due to social factors. CONCLUSIONS: This study has demonstrated the lasting and greater effect of length and weight at birth on height in comparison to social factors, but social factors are still meaningful in their effects on height. Birth length has a marginal greater effect on final height than birth weight. PMID- 12573080 TI - Social status and adult heights in the two Germanies. AB - BACKGROUND: Lack of evidence prevented hitherto the systematic comparison of physical stature across East and West Germany. AIM: The aim of this study is to compare heights by social status in the two Germanies with very different socio economic and political systems prior to unification. DATA: The German Federal Health Survey of 1998 (Public Use File BGS98), a cross-sectional random sample of 7124 adult males and females between ages 18 and 79 (birth cohorts of 1919-1980) is used in the analysis. RESULTS: There are considerable and persistent differences by social status in both East and West Germany over time. The West German height advantage among men increases with social status, whereas among women the opposite is the case. East German men born after the Berlin Wall was built became significantly shorter, but East Germans regained equality with West Germans after unification. In contrast, East German females were markedly shorter than their West German counterparts throughout the period considered. CONCLUSIONS: The West German capitalistic welfare state provided a more propitious environment for the physical growth of the human organism than did the socialist East German government. There were substantial social differences in height in the officially classless German Democratic Republic. PMID- 12573081 TI - The prediction of height by armspan in older Chinese people. AB - OBJECTIVE: To compare the accuracy of the predictive equations derived from older Chinese people without vertebral deformity with those derived from unselected older people, in estimating height from armspan. METHODOLOGY: Height and armspan were measured in 789 older Chinese and 885 young Chinese in Hong Kong. All the older subjects had spinal X-ray (T4 to L4). Vertebral deformity was determined by a trained technician using a translucent digital analyser. Definite deformity was defined by a greater than 3 standard deviation increase in one of the three vertebral height ratios (anterior/posterior, middle/posterior and posterior/posterior). Eighty of the older subjects without vertebral deformity were randomly selected from the studied population for internal validation of predictive equations. The remaining subjects were used to derive sex-specific predictive equations for height by linear regression against armspan. The accuracy of these equations was analysed by the Bland and Altman method. RESULTS: Vertebral deformity was present in 30.7% and 15.6% of older female and male subjects, respectively. Older women without vertebral deformity had significantly greater armspan: height ratio when compared with the young (1.04 versus 1.02), but the corresponding ratios were the same in older and young men (both 1.04). The predictive equation derived from older females without vertebral deformity had lower standard error of estimation and gave estimates closer to measured height than those derived from unselected older women. But this was not the case in older men. CONCLUSION: In deriving predictive equation to estimate height by armspan in older Chinese women, selecting out older women without vertebral deformity increased accuracy of the equation. This was, however, not the case in older men. PMID- 12573082 TI - Cohort effects in a genetically determined trait: eye colour among US whites. AB - BACKGROUND: While the inheritance of eye colour is likely polygenic, blue eye colour is thought to follow an inheritance pattern similar to that of a recessive trait. Consequently, age-related differences in the prevalence of blue eye colour would be unanticipated. AIM: This study explores the finding and explanation for birth cohort differences in the prevalence of blue eye colour in the US white population. SUBJECTS AND METHODS: Data from the first (1971-1975) and third (1988 1994) US National Health and Nutrition Examination Surveys (NHANES-I and NHANES III), nationally representative surveys of the US population, were analysed. Trends in eye colour prevalence by birth cohort were analysed together with mortality rates according to eye colour. US census data (1980) were examined to explore cohort differences in ancestry and assortative mating by ancestry. RESULTS: The prevalence of blue eye colour among non-Hispanic whites in NHANES III was 57.4% (95% CI: 50.1-64.7) for individuals born between 1899 and 1905 compared to 33.8% (95% CI: 31.3-36.5) for those born between 1936 and 1951. No association was found between survival and eye colour, nor was a cohort effect evident for primary ancestry. However, proportions reporting only one ancestry in census data declined with successive birth cohorts. CONCLUSIONS: A cohort effect in blue eye colour prevalence was found for the US white population. A secular trend of decreasing assortative mating by ancestry is the likely explanation. PMID- 12573083 TI - Growth tracks in pre-pubertal children. AB - The present investigation characterizes common growth tracks in pre-pubertal children. Growth tracks denominate areas of probability within which subsequent measurements of the body height (or body height SDS) of a healthy individual will predominantly be found. Growth tracks are defined over several years and they are insensitive to the timing of measurements. The concept of growth tracks was developed to improve separating aberrant patterns from normal growth. Longitudinal data on height were obtained from six large national growth studies, performed at Berkeley, USA, Jena, Germany, Lublin, Poland, Paris, France, Prague, Czech Republic and Zurich, Switzerland with a total of 515 healthy boys and 532 healthy girls. Four hundred and two series of annual height measurements were available in pre-pubertal boys (aged 3-11 years), 416 series in pre-pubertal girls (aged 3-10). Body height was converted into height SDS. Thereafter, average personal height SDS was determined, and subtracted from height SDS, resulting in individual series of residual height SDS. These were sorted by cluster analysis and distributed into groups (clusters) according to similarity or dissimilarity (squared difference). We identified similar clusters, and named them 'growth tracks'. We found five pre-pubertal male growth tracks, each containing between 4 and 37% of the boys. Twenty boys could not be assigned to either one of the five tracks. Very similar results were obtained in girls, with five pre-pubertal growth tracks also, each containing between 3 and 50%. Twenty-three individuals grew irregularly and could not be assigned. Growth tracks are narrow, with an average width between 12.1 and 14.8% of the SD of body height. Most children exhibited almost horizontal height SDS patterns. Others showed linearly declining, rising, or intersection -shaped patterns. None of the patterns were predominantly found in particularly short or tall children. Preliminary data support the practical advantages of the concept of growth tracks. PMID- 12573084 TI - Population genetic study of the STR loci (HUMCSF1PO, HUMTPOX, HUMTHO1, HUMLPL, HUMF13A01, HUMF13B, HSFESFPS and HUMVWA) in North Indians. AB - BACKGROUND: Highly polymorphic genetic markers like short tandem repeats (STRs) have been used successfully in disease analysis and studies of human evolution and population genetic diversity. However, DNA-based population genetic studies of Indian populations are limited. SUBJECTS AND METHODS: To enlarge our understanding of genetic variation in Indian populations, a population genetic study was carried out on Jat Sikh (Punjab, North India) individuals (n = 150) using a battery of the STR loci. The STR loci analysed by means of PCR amplification followed by electrophoresis and silver staining included HUMCSF1PO, HUMTPOX, HUMTHO1, HUMLPL, HUMF13A01, HUMF13B, HUMFESFPS and HUMVWA loci. RESULTS: The overall pattern of allele frequencies was similar to many Caucasian and Indian populations and heterozygosity varied from 65% (HUMLPL) to 85% (HUMVWA). For all eight loci, no deviations from the Hardy-Weinberg equilibrium hypothesis were detected. Significant differences were observed between Jat Sikhs and African, Chinese and Indian tribes. The mean exclusion probability ranged from 35% to 70%, and the power of discrimination from 81% to 93%, indicating the potential of these loci for forensic and paternity investigations. CONCLUSION: The allele frequency spectrum, heterozygosity, probability of exclusion, match probability and discrimination probability estimates show interesting variation and suggest the usefulness of these loci for anthropogenetic, paternity and forensic investigations in Indian populations. PMID- 12573085 TI - Consanguineous marriage in PR China: a study in rural Man (Manchu) communities. AB - Although there is a long history of consanguineous marriage in China, information on its prevalence is very limited. The Man (Qing) dynasty ruled China for over 250 years, but no consanguinity studies have been reported on this important population. The objective of the present investigation was to determine the present-day level of consanguineous marriage in the Man community, and to compare the data with existing consanguinity information on other Chinese populations. The study was conducted in a group of 11 rural Man communities in the north eastern Chinese province of Liaoning. Household-based interviews were conducted by local staff on 513 couples, 418 of whom were Man with another 95 Man-Han inter ethnic marriages. Basic pedigrees were constructed to determine the biological relationship between each set of spouses. Thirty of the 418 couples were in a consanguineous union, with a mean coefficient of inbreeding alpha = 0.0012. The small population sizes of the study may have contributed to the spatial variation in the patterns of inbreeding. Across generations there was a reduction in consanguineous marriages and an increase in inter-ethnic unions, which paralleled changes in civil marriage regulations. PMID- 12573104 TI - Isolation of native human monoclonal autoantibodies to breast cancer. AB - Using a unique fusion partner cell line, MFP-2, and B-lymphocytes from breast cancer patients, we developed a set of fully human monoclonal antibodies (MAbs) that bind with high specificity and sensitivity to breast cancer cells. Immunofluorescent staining of normal tissues, primary tumors, and metastatic lymph nodes demonstrates that these antibodies are specific for breast cancer of autologous and allogeneic origin. We have also determined that many of the antibodies selected based on specific binding to breast cancer cells and tissue also bind prostate cancer cells and tissue with high specificity and sensitivity. The targets of these antibodies have been localized to the cytoplasm and membrane. Biological assays for internalization and cytotoxicity demonstrated the ability of three antibodies to rapidly internalize. Our study demonstrates that isolation of native human MAbs from the natural antibody repertoire, targeted to cancer cells, is feasible and may provide a source of tools for immunotherapy. PMID- 12573105 TI - Genetic engineering of streptavidin-binding peptide tagged single-chain variable fragment antibody to Venezuelan equine encephalitis virus. AB - A recombinant gene encoding a single-chain variable fragment (scFv) antibody against Venezuelan equine encephalitis virus (VEE) was cloned into a prokaryotic T7 RNA polymerase-regulated expression vector. A streptavidin-binding peptide gene fused to a 6His tag was attached downstream to the scFv gene. The recombinant fusion protein was expressed in bacteria as inclusion bodies that were subsequently solubilized with 8 M urea and renatured by an arginine system. Purification of the fusion protein was achieved by immobilized metal affinity chromatography. Enzyme-linked immunosorbent assay (ELISA) and Western blotting results revealed that the fusion protein not only retained VEE antigen binding and specificity properties similar to those of its parent native monoclonal antibody (MAb), but also possessed streptavidin-binding activity. This experimental approach can eliminate the need for chemical biotinylation of antibodies and the risk associated of antibody denaturation and can provide a stable and reproducible reagent for rapid and efficient immunoassay of VEE when detected by horseradish peroxidase (HRP)-conjugated streptavidin. PMID- 12573106 TI - Generation of human interferon gamma and tumor Necrosis factor alpha chimeric TNT 3 fusion proteins. AB - Studies have shown that cytokines can effectively treat solid tumors by a direct cytotoxic effect as well as by immunomodulation. Both human interferon gamma (IFNgamma) and tumor necrosis factor alpha (TNFalpha) have been used to treat a variety of colon carcinoma cell lines and tumors in patients. These cytokines, however, are dose limited by their toxicity and fast clearance rates when given intravenously. To improve their therapeutic value, we now report on the generation of two new fusion proteins consisting of human IFNgamma and TNFalpha genetically linked to the C-terminal portion of chTNT-3, a monoclonal antibody (MAb), which targets human solid tumors by binding to intracellular antigens exposed in degenerating cells associated with tumor necrosis. In vitro characterization studies demonstrate that both the IFNgamma and TNFalpha fusion proteins are able to maintain their binding affinity to antigen as well as their direct cytotoxic effect and immunomodulatory functions. When both fusion proteins are combined at optimal doses, they demonstrate a 30% direct cellular cytotoxicity of human colon carcinoma cells of which approximately 14% can be attributed to apoptosis. In vivo, these agents were studied for their pharmakocinetic clearance rates and their ability to target human colon carcinomas heterotransplanted in nude mice. The results of these studies show that, compared with chTNT-3 parental antibody, both fusion proteins have a substantially shorter whole body half-life, yet are able to target tumor in a similar manner. As each of these fusion proteins are cleared from the circulation and normal tissues, tumor-to-normal-tissues ratios rise demonstrating the retention of these reagents in tumor. The generation of long-acting and targeted human IFNgamma and TNFalpha antibody fusion proteins will enable investigators to study the role of these potent immunostimulatory cytokines in the treatment of human solid tumors. PMID- 12573107 TI - Anti-idiotypic antibody as the surrogate antigen for cloning scFv and its fusion proteins. AB - Single-chain variable fragment (ScFv) is a versatile building block for novel targeting constructs. However, a reliable screening and binding assay is often the limiting step for antigens that are difficult to clone or purify. Anti idiotypic antibodies may be useful as surrogate antigens for cloning scFv and their fusion proteins. 8H9 is a murine IgG(1) monoclonal antibody (MAb) specific for a novel antigen expressed on the cell surface of a wide spectrum of human solid tumors, but not in normal tissues. Rat anti-8H9-idiotypic hybridomas (clones 2E9, 1E12, and 1F11) were produced by somatic cell fusion between rat lymphocytes and mouse SP2/0 myeloma. In direct binding assays enzyme-linked immunosorbant assay--(ELISA)--they were specific for the 8H9 idiotope. Using 2E9 as the surrogate antigen, 8H9-scFv was cloned from hybridoma cDNA by phage display. 8H9scFv was then fused to human-gamma1-CH2-CH3 cDNA for transduction into CHO and NSO cells. High expressors of mouse scFv-human Fc chimeric antibody were selected. The secreted homodimer reacted specifically with antigen-positive tumor cells by ELISA and by flow cytometry, inhibitable by the anti-idiotypic antibody. The reduced size resulted in a shorter half-life in vivo, while achieving comparable tumor to nontumor ratio as the native antibody 8H9. However, its in vitro activity in antibody-dependent cell-mediated cytotoxicity was modest. PMID- 12573108 TI - Characterization of a pan-species reactive monoclonal antibody specific for a cell surface epitope which could serve as a marker for human monocytic and megakaryocytic differentiation. AB - A mouse IgG(1)-producing hybridoma, CSC-31, was isolated and characterized. The monoclonal antibody (MAb) was originally raised against monkey kidney cell surface molecules. FACS analysis further showed that CSC-31 exhibited broad tissue and species reactivity. Although most human T- and B-cell lines failed to react with CSC-31, myeloid, and erythroleukemia cells lines such as THP-1 and K562 expressed the CSC-31 cell surface marker. Furthermore, in vitro differentiation of HL-60, U-937, and K562 showed that expression of the CSC-31 marker is associated with monocytic or megakaryocytic differentiation. However, up-regulation of the CSC-31 marker expression was not detected during granulocytic or erythroid differentiation. Through the in vitro differentiation of K562, it was demonstrated that up-regulation of the CSC-31 marker required novel PKCs and might be regulated by the MAPK signaling pathway. Last, limited biochemical analysis demonstrated that the CSC-31-specific epitope is sensitive to digestion by papain yet highly resistant to other proteases. PMID- 12573109 TI - Monoclonal antibodies KN-02 and KN-03 against the heavy chain of kinesin. AB - The present paper describes two new monoclonal antibodies (MAbs) KN-02 and KN-03 against the heavy chain of conventional kinesin. The kinesin was purified from porcine brain by a combined procedure of ion exchange chromatography, tripolyphosphate-supported microtubule affinity-binding, and gel filtration. Hybridoma cell lines producing antibodies were obtained after immunization of a Balb/c mouse with kinesin and subsequent fusion of the spleen cells with Sp2/0 myeloma cells. The specificity was verified by enzyme-linked immunosorbent assay (ELISA) and further confirmed by immunoblotting and immunoprecipitation analysis. The antibodies recognize different epitopes on the heavy chain of the kinesin molecule as demonstrated by chymotryptic cleavage of kinesin followed by immunoblotting. Differential location of relevant epitopes was also documented by in vitro binding experiments with purified kinesin and taxol-stabilized microtubules. While the KN-03 antibody decorated microtubules, no such staining was observed with KN-02 antibody. The antibodies have a lower affinity to sodium dodecyl sulfate (SDS)-denatured kinesin, but immunofluorescence on fixed cells gave strong dot-like staining characteristic for localization of kinesin on vesicles. The same staining pattern was observed in different cell types. Double label fluorescence with polyclonal anti-tubulin antibody revealed a co distribution of stained vesicles with microtubules on the cell periphery. The antibodies KN-02 and KN-03 are therefore valuable tools for localization of kinesins in cells of different tissue origin. PMID- 12573110 TI - In vivo and in vitro anti-tumor effect of 14F7 monoclonal antibody. AB - The 14F7 monoclonal antibody (MAb) is an IgG(1) antibody that reacts specifically with GM3 (NeuGc) and with tissue sections of human tumors. We demonstrated here that this MAb is agglutinin that specifically agglutinated horse erythrocytes. Additionally, the capacity of 14F7 MAb to mediate cytotoxicity against GM3 (NeuGc)-positive murine myeloma cells, in vitro and in vivo, was evaluated. High concentrations of 14F7 MAb were needed to induce complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) against the murine myeloma cells. The most relevant finding was the ability of this MAb to directly kill the target cells without participation of complement. This cytotoxicity was dependent on the temperature and MAb concentration and the number of the target cells. In vivo, the passive treatment with 14F7 MAb produced a strong anti-tumor activity, similar to the anti-tumoral response obtained with standard chemotherapy treatment. PMID- 12573112 TI - Production and characterization of monoclonal antibodies against asexual stages of Plasmodium yoelii nigeriensis. AB - Swiss mice vaccinated with Plasmodium yoelii nigeriensis-soluble antigen and saponin, following a homologous 100% lethal challenge, showed 60% protection (6 out of 10 mice survived). Monoclonal antibodies (MAbs), generated by hybridizing the Sp2/0 myeloma cells with the splenocytes of each of these ten mice, separately, were screened using enzyme-linked immunosorbent assay (ELISA), and were characterized by using merozoite (Mz) invasion inhibition assay in vitro, immunofluorescence assay (IFA), passive transfer of protection and ELISA-based isotyping. Curiously, purified MAbs from each of the six protected mice showed a distinct dichotomy: only two or three of them inhibited >86% Mz invasion, whereas the remaining six to nine showed <58% Mz invasion inhibition. However, none of the purified MAbs from the nonprotected mice could inhibit >58% Mz invasion. Furthermore, the ability of the MAbs to inhibit Mz invasion appeared to correlate with their IFA-reactivity with the free-Mz, suggesting that these MAbs were directed against the Mz surface antigens involved in invasion. In passive transfer of protection experiments, pooled purified MAbs from protected mice, that inhibited >86% Mz invasion, transferred 60% protection from challenge; the remaining pooled purified MAbs from protected mice, and those from nonprotected mice, when transferred separately, imparted only 30 and 10% protection, respectively. Isotypically, the MAbs belonged to IgG(1), IgG(2a), IgG(2b), and IgG(3) subclasses. Our results indicate that purified MAbs against P. yoelii nigeriensis, produced from the hybrids generated using the splenocytes of vaccinated and protected mice, belonged to two distinct groups: a small group that inhibited >86% Mz invasion, strongly cross-reacted with free-Mz, transferred up to 60% passive protection, and belonged to IgG(2a) and IgG(3) subclasses, whereas the other relatively larger group inhibited <58% Mz invasion, weakly cross-reacted with free-Mz, and transferred only 30% passive protection. PMID- 12573111 TI - Production and characterization of species-specific monoclonal antibodies against Actinomyces naeslundii and Lactobacillus casei. AB - Dental caries is a disease induced by a few cariogenic bacterial species. Quantitative detection of these cariogenic bacteria could provide useful information for caries risk assessment. In this study, we developed highly species-specific monoclonal antibodies (MAbs) against the type strains of Actinomyces naeslundii ATCC 12104 and Lactobacillus casei ATCC 11578. Assayed with immunoprecipitation and immunofluorescent microscopy, these antibodies showed high sensitivities and specificities in detecting A. naeslundii and L. casei in saliva. Examining 100 saliva samples using these MAb-based techniques, we found that the number of A. naeslundii in saliva ranges from 0.5 x 10(4) to 4.8 x 10(5) and that the number of L. casei in saliva ranges from 1 x 10(4) to 1.2 x 10(6). With fluorescent imaging techniques and confocal laser scanning microscopy (CLSM), these antibodies were used to visualize A. naeslundii and L. casei within dental plaques derived from stimulated whole human saliva in vitro. The study demonstrated that these MAbs were also able to effectively detect A. naeslundii and L. casei in plaque. PMID- 12573113 TI - Monoclonal antibody against human trypsin: production, characterization, and use for diagnosis. AB - A monoclonal antibody (MAb) directed against human immunoaffinity purified trypsinogen has been produced by hybridization of myeloma cells with spleen cells of BALB/c immunized mice. Antibodies were screened by ultramicro-enzyme-linked immunosorbent assay (UMELISA). The MAb was purified by affinity chromatography on protein A-sepharose, and MAb had a high affinity for trypsinogen with the affinity constant equal 2.06 x 10(9) L/mol. Specificity was studied by UMELISA using cross-reactant proteins; MAb gave a positive reaction with native trypsinogen-1 but did not react with the same protein after reduction. The antibody seem to be directed against conformational epitope. The MAb obtained was characterized immunologically and used to develop UMELISA for detection Trypsin. This monoclonal assay enabled the detection of 2.8 ng/mL. PMID- 12573135 TI - Angiotensin II induces a rapid and transient increase of reactive oxygen species. AB - Vascular smooth muscle cells (VSMC) exhibit a hypertrophic and contractile response after angiotensin II (Ang II) treatment, and the NADH/NADPH oxidase dependent synthesis of hydrogen peroxide (H(2)O(2)) seems to play a central role in these responses. Present experiments were designed to analyze the mechanisms responsible for the rapid changes induced by Ang II in the intracellular H(2)O(2) concentration in VSMC. Ang II induced a quick and transient increase of dichlorodihydrofluorescein (DCHF) fluorescence in VSMC, an effect that was completely abolished by catalase and by diethyldithiocarbamate, a cell-permeable superoxide dismutase inhibitor. Losartan and pertussis toxin prevented the stimulatory effect of Ang II. Both diphenylene iodonium (NADH/NADPH oxidase blocker) and 3-(4-octadecylbenzoyl)acrylic acid (phospholipase A2 blocker) inhibited the changes in DCHF fluorescence induced by Ang II, in a dose-dependent fashion, and the effects of both inhibitors were additive. These data demonstrate that Ang II induces a very quick and transient increase of H(2)O(2) in VSMC. This effect depends on the receptor type 1, is linked to a G protein, and involves both NADH/NADPH oxidase and phospholipase A2 activation. The mechanism may be related to the previously proposed role of H(2)O(2) in the genesis of the Ang II induced cell contraction. PMID- 12573136 TI - PPAR-alpha ligands inhibit H2O2-mediated activation of transforming growth factor beta1 in human mesangial cells. AB - Transforming growth factor-beta1 (TGF-beta1) mediates the development of glomerulosclerosis by stimulating mesangial cell production of extracellular matrix (ECM) proteins. TGF-beta1 and several ECM genes are regulated by promoter O-tetradecanoylphorbol 13-acetate-responsive elements (TREs) that are transactivated by the activator protein-1 (AP-1) transcription factor complex. AP 1-TRE interactions are regulated by redox changes. Recently, peroxisome proliferator-activated receptors (PPARs) were shown to negatively regulate several transcription factor families. In these studies, we postulated that PPAR alpha could antagonize TGF-beta1 expression by cultured human mesangial cells (HMC). A TGF-beta1 luciferase expression plasmid was transduced into HMC via recombinant deficient adenoviral vectors. The TGF-beta1 promoter activity increased twofold (209%) following 18-h treatments with H(2)O(2) (1,000 micro M). Using RT-PCR, we demonstrated that HMC possess PPAR-alpha RNA, and PPAR-alpha protein was identified by immunohistochemistry. Pretreatment of cells with the PPAR-alpha ligands WY14643 (100-500 micro M) or clofibrate (100-500 micro M) dose dependently inhibited oxidant-mediated induction of TGF-beta1. This inhibition occurred without affecting the H(2)O(2)-mediated activation of the mitogen activated protein kinase (MAPK) pathways extracellular regulated kinase, p38 MAPK, or Jun N-terminal kinase, which are responsible for the regulation of AP-1 phosphorylation. These studies are the first to identify PPAR-alpha expression by HMC. The results of these studies suggest that TGF-beta1 expression mediated by oxidant stress may be suppressible by PPAR-alpha activation. PMID- 12573137 TI - Renal dysfunction after chronic blockade of nitric oxide synthesis. AB - The effects of the chronic inhibition of nitric oxide (NO) on renal hemodynamics and tubular function were studied in rats treated for 8 weeks with the NO synthesis inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME; 40 mg/kg/day). In addition, the effect of L-NAME administration on vasoactive systems (renin angiotensin system, aldosterone, catecholamines, endothelin, and thromboxane A(2)) was evaluated. Chronic inhibition of NO significantly elevated blood pressure, reduced glomerular filtration rate and renal blood flow, blunted the pressure-diuresis-natriuresis response, and increased protein urine excretion. All these changes were associated with blunted nitrite production in response to acetylcholine in glomeruli. No changes were observed in the plasma levels of either renin activity, aldosterone, or endothelin in L-NAME-treated rats. Similarly, no differences were observed in the urinary excretion of thromboxane B(2) between both group of animals. By contrast, plasma concentrations of both epinephrine and norepinephrine were elevated in rats treated with L-NAME. In summary, the results show that chronic blockade of NO produced not only alterations in renal function, but also renal damage, suggesting an important renoprotective role of NO. An activation of sympathoadrenal system could participate in these renal alterations. PMID- 12573138 TI - Protective effect of trans-resveratrol on gentamicin-induced nephrotoxicity. AB - Reactive oxygen species (ROS) have been involved in glomerular filtration rate (GFR) reduction observed after gentamicin treatment. trans-Resveratrol (TR), a natural hydroxystilbene, has been identified to be a potent inhibitor of ROS production. The aim of this work has been to study whether TR has a protective effect on gentamicin-induced nephrotoxicity in vivo and the effect of TR on lipid peroxidation and the oxidative stress induced by gentamicin. Animals that received a daily intraperitoneal injection of gentamicin (100 mg/kg body weight) showed lower GFR and renal blood flow (RBF) and higher urinary excretion of N acetyl-beta-D-glucosaminidase (NAG) than control rats. Rats receiving TR together with gentamicin showed higher GFR and RBF and lower NAG urinary excretion than rats receiving gentamicin alone. Moreover, renal lipid peroxidation increased in rats receiving gentamicin alone, and this increase was prevented by the administration of TR. The concentration in plasma of antioxidants was higher in the group that received TR with gentamicin than in the gentamicin and control groups. The activities of lactate dehydrogenase and alkaline phosphatase were higher in rats treated with gentamicin than in control rats and were reduced by the treatment with TR. This study demonstrates an improvement in renal function in response to the administration of TR in gentamicin-induced nephrotoxicity. At least a part of this effect of TR could be based on its antioxidant activity. PMID- 12573139 TI - NAD(P)H oxidase-derived reactive oxygen species as mediators of angiotensin II signaling. AB - Angiotensin II has been shown to participate in both physiological processes, such as sodium and water homeostasis and vascular contraction, and pathophysiological processes, including atherosclerosis and hypertension. The effects of this molecule on vascular tissue are mediated at least in part by the modification of the redox milieu of its target cells. Angiotensin II has been shown to activate the vascular NAD(P)H oxidase(s) resulting in the production of reactive oxygen species, namely superoxide and hydrogen peroxide. In this article, we review what is known about the molecular steps that link angiotensin II and its receptor to production of reactive oxygen species and subsequent redox mediated events, focusing on the structural and functional properties of the vascular NAD(P)H oxidases and their downstream mediators. As such, we provide a framework linking angiotensin II to crucial vascular pathologies, such as hypertension, atherosclerosis, and restenosis after angioplasty, by means of the NAD(P)H-dependent oxidases and their effector molecules. PMID- 12573140 TI - Apoptotic pathways of oxidative damage to renal tubular epithelial cells. AB - Toxic renal failure induced by gentamicin, glycerol, or cisplatin, as well as ischemic renal failure in vivo and hypoxia/reoxygenation of tubular epithelial cells in vitro, induces the production of reactive oxygen metabolites (ROM). Generation of ROM is responsible for the induction of tubular epithelial cell death, which is mediated by caspases and/or endonucleases. Scavenging of ROM protects tubular epithelium from caspase and endonuclease activation and from cell death. Thus, the inhibition of ROM production combined with the pharmacological control of caspase and endonuclease pathways may provide future modalities in the prevention or treatment of acute renal failure in humans. PMID- 12573141 TI - Acute renal failure: is nitric oxide the bad guy? AB - Nephrotoxicity is a major side effect in clinical practice, frequently leading to acute renal failure (ARF). Many physiological mechanisms have been implicated in drug-induced renal injury. Currently, nitric oxide (NO) is considered to be an important regulator of renal vascular tone and a modulator of glomerular function under both basal and physiopathological conditions. Historically, NO has been implicated in ARF and, after its discovery, several publications have suggested that changes in NO production could play an important role in the hemodynamic alterations observed in ARF. In this review, we evaluate the participation of NO in ARF and summarize many of the findings in this research area in an attempt to elucidate the role of NO in ARF. PMID- 12573142 TI - Evolution of oxidative stress and inflammation during hemodialysis and their contribution to cardiovascular disease. AB - End-stage renal disease patients have increased cardiovascular morbidity and mortality. These patients have many unique risk factors, such as an accumulation of uremic toxins, electrolyte imbalances, metabolic disturbances, anemia, chronic inflammation, and thrombogenic disturbances. Oxidative stress has been implicated in many of these disturbances. This review will focus on some of the factors that may accelerate cardiovascular disease in uremic patients, with an emphasis on mechanisms and interactions of various components of oxidative stress and inflammation. Understanding the mechanisms of these pathways may be useful in developing effective prevention and treatment strategies. PMID- 12573143 TI - Regulation of the activation of nuclear factor kappaB by mitochondrial respiratory function: evidence for the reactive oxygen species-dependent and independent pathways. AB - Mitochondrial respiratory function regulates the redox status of cells, which, in turn, can control the activation of transcription factors. However, how mitochondria accomplish this modulation is not completely understood. Using the human myelogenous leukemia cells ML-1a, respiration-deficient clone 19 derived from ML-1a, and reconstituted clones, we demonstrated the role of respiratory function in the activation of nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1). Constitutive activation of NF-kappaB and AP-1 was observed in clone 19, but not in ML-1a, and the constitutive activation observed in clone 19 was completely inhibited in reconstituted clones that have functional mitochondria. Additionally, tumor necrosis factor (TNF)-induced activation of NF kappaB and AP-1 observed in ML-1a was greatly reduced in clone 19. These results indicate that mitochondrial respiratory function regulates TNF-induced and constitutive activation of NF-kappaB and AP-1. We investigated the roles of reactive oxygen species in NF-kappaB activation. Generation of superoxide detected by hydroethidine, but not hydrogen peroxide detected by dehydrorhodamine 123, was transiently increased by TNF in both of the cells. The antioxidant, pyrrolidine dithiocarbamate, reduced TNF-induced, but not the constitutive, NF kappaB activation. These results indicate that the increase in superoxide generation might be involved in TNF-induced, but not in constitutive, NF-kappaB activation. Our results thus demonstrate the involvement of mitochondrial respiratory function in the activation of reactive oxygen species-dependent and independent pathways for NF-kappaB activation. PMID- 12573168 TI - Harm reduction or harm maintenance? PMID- 12573170 TI - Tobacco harm reduction: a call to address the ethical dilemmas. AB - The 2001 Institute of Medicine report Clearing the Smoke: Assessing the Science Base for Tobacco Harm Reduction has helped to focus attention on the scientific basis for assessing tobacco harm reduction products. As the tobacco research and policy communities tackle the challenges of evaluating harm reduction, there are ethical issues that must also be addressed. There has, however, been very little writing on the ethics of this field. In an effort to spur research into answering these ethical questions, we present two complementary approaches. First we outline three overarching topics in tobacco harm reduction that would particularly lend themselves to study: (a) Is the pursuit of tobacco harm reduction an ethical goal? (b) What are the ethical considerations of tobacco harm reduction vis-a-vis pharmaceutical companies? and (c) What are the ethical considerations for harm reduction vis-a-vis tobacco companies? We then present one possible framework for analyzing the ethical issues that accompany particular tobacco harm reduction strategies. By considering the ethical dilemmas attendant to tobacco harm reduction in a prospective and thoughtful manner, we will be better prepared to handle the challenges that face us individually as researchers and collectively as a tobacco control community. PMID- 12573169 TI - Nicotine concentrations with concurrent use of cigarettes and nicotine replacement: a review. AB - This paper reviews the data on blood nicotine or saliva cotinine concentrations with concomitant smoking and use of nicotine replacement (NR) products. Eleven studies that provided data on blood nicotine concentrations, carbon monoxide in exhaled air, and number of cigarettes smoked were reviewed. At least one day had to be spent on concurrent use of cigarette and NR products. With simultaneous use of smoking and acute NR products (gum and inhaler) the nicotine concentrations were unchanged, whereas they increased (+54%) with nicotine patches. With both types of NR products, the number of cigarettes smoked per day was reduced by approximately 50% and carbon monoxide (CO) by 30%. Where smokers had the intention or received instructions to reduce smoking, a greater reduction in cigarettes smoked and exhaled CO was observed. Despite substantially increased nicotine concentrations (e.g., up to 3 times the approved dose) there were no significant adverse reactions. Concurrent use of NR products and cigarette smoking appears to be safe. PMID- 12573171 TI - Reducing tobacco harm: research challenges and issues. AB - The emergence of potential reduced-exposure tobacco and cigarette-like products and the reduction of smoking as a treatment approach, have recently been forcing the debate and discussion about the science that is necessary to inform policies, regulation, and programs. To deal effectively with the issues evolving around tobacco harm reduction, a comprehensive and strategic research agenda must be forged, and a multi-disciplinary, collaborative approach must be taken. The goal of this article is to describe research challenges and issues related to tobacco exposure and harm reduction. Scientists from multiple disciplines and individuals from government agencies, nonprofit organizations, and tobacco control advocacy groups as well as the pharmaceutical and tobacco industries attended a two-day meeting that focused on addressing the current knowledge regarding tobacco harm reduction, identifying gaps in knowledge, and recommending research directions. Workgroups, comprising a subset of attendees, met after the conference to synthesize, discuss, and prioritize the important research frontiers. The resulting document provides guidance for scientists, grant funding agencies, industry and policy makers by identifying areas in which to invest research effort and funds to develop a science base to ensure the future health of this nation and world. PMID- 12573172 TI - Light smokers: issues and recommendations. AB - Although the overall prevalence of tobacco smoking in the United States is decreasing, the proportion of light smokers is increasing and is as high as 50% among some groups. There is growing evidence that light smokers experience significantly higher health risks compared to nonsmokers, and they are often unable to quit smoking on their own. The prevalence of light smoking differs among subpopulations in the US and is particularly common among adolescents, college students, pregnant women, and some ethnic minorities. Characteristics of light smokers in these subpopulations vary, as do the underlying reasons for smoking, barriers to cessation, and treatment issues. Unfortunately, the paucity of research to develop and test effective cessation aids is common across all groups of light smokers. Given the substantial portion of light smokers in these populations, development and dissemination of effective behavioral and pharmacological interventions for light smokers is likely to result in substantial smoking reduction rates in the general population. PMID- 12573173 TI - Tobacco harm reduction: conceptual structure and nomenclature for analysis and research. AB - The goal of tobacco control has always been to reduce death and disease due to tobacco use. Recent discussions have broadened the concept of tobacco control beyond cessation and prevention to include concepts such as the use of medications to achieve reduction in tobacco use, chemoprevention to reduce disease, modifications of tobacco products to reduce toxicity, and behavioral approaches to change smoking and tobacco use behavior. Within each of these broad domains, diverse approaches have been suggested. To facilitate clear discussion and analysis, and to avoid confusion among approaches, we catalog 19 approaches to harm reduction, distinguishing and discussing them on 11 dimensions, including their objectives, mechanisms, toxicology, expected population risks, and consumer appeal. Because there have also been so many suggested applications of medicinal nicotine to smoking intervention, we separately catalog and analyze nine applications, some of which constitute new approaches to harm reduction. The suggested framework is intended to clarify the debate, provide for common nomenclature, and facilitate analysis of diverse approaches to tobacco harm reduction. PMID- 12573174 TI - Evaluating acute effects of potential reduced-exposure products for smokers: clinical laboratory methodology. AB - Harm reduction for tobacco smokers may involve reducing their exposure to lethal smoke constituents. Assessing smoke constituent exposure and any resulting harm reduction from a potential reduced-exposure product (PREP) will involve preclinical, clinical, and epidemiological research. The purpose of this study was to evaluate a clinical laboratory model for assessing the acute effects of PREPs for smokers. Philip Morris' Accord and R.J. Reynolds' Eclipse were used as examples. Twenty overnight-abstinent smokers (> 15 'light' or 'ultra-light' cigarettes/day) participated in 4 Latin-square ordered, 2.5-hr sessions in which they completed an 8-puff smoking bout every 30 minutes. Sessions were separated by at least 24 hours and differed by product used: own brand, denicotinized tobacco cigarettes, Accord, or Eclipse. Tobacco withdrawal and carbon monoxide (CO) were assessed before and after smoking, heart rate was assessed before and during smoking, and puff volume, duration, and interpuff interval were assessed while subjects smoked. Blood was sampled at the beginning and end of each session. Relative to normal cigarettes, Accord was less effective at suppressing withdrawal and produced minimal CO boost despite the fact that, when using Accord, subjects took bigger and longer puffs. Eclipse suppressed withdrawal fully and increased CO boost by approximately 30%. Own brand, Accord, and Eclipse, but not denicotinized cigarettes, increased plasma nicotine concentration. Taken together, these results suggest that neither Accord nor Eclipse is likely to be an effective reduced-exposure product for smokers and that this clinical laboratory model is valuable. PMID- 12573175 TI - Long-term effects of the Eclipse cigarette substitute and the nicotine inhaler in smokers not interested in quitting. AB - This study investigated the long term use of Eclipse, a cigarette-like device that heats rather than burns tobacco, and the Nicorette oral inhaler, a nicotine replacement product, in a group of smokers not interested in quitting smoking. Subjects were recruited from a previous four-week controlled trial with the Eclipse and the inhaler (Fagerstrom, Hughes, Rasmussen, & Callas, 2000). Thirty eight subjects from the previous study were given the opportunity to continue to reduce their smoking for eight weeks and to choose Eclipse or the inhaler to aid reduction in cigarette smoking. If they did not like either of the two products they were allowed to smoke their own brand of cigarettes. A total of 39% percent chose to use the inhaler, 26% the Eclipse, and 34% neither. Number of cigarettes smoked per day decreased 86% with the Eclipse and 68% with the inhaler. Exhaled carbon monoxide increased 45% with Eclipse and decreased 47% with the inhaler. No differences were seen with craving for cigarettes between the products. Cigarette users showed stable values for cigarettes per day, exhaled carbon monoxide (CO) and nicotine/cotinine concentrations. These results replicate and extend our prior results; i.e. both Eclipse and inhaler clearly reduced smoking of traditional cigarettes and with no loss of effect over time. For Eclipse this was accompanied by maintenance of nicotine levels but increases in CO whereas for the inhaler this was accompanied by a reduction in CO intake. PMID- 12573176 TI - Does tobacco industry marketing of 'light' cigarettes give smokers a rationale for postponing quitting? AB - The objective of this analysis was to examine further whether tobacco industry marketing using the labels light and ultra-light is perceived by smokers as a health claim. Smokers might view low tar/nicotine brands of cigarettes as a means to reduce the harm to their health from smoking and postpone quitting. Data were from smokers responding to a large, population-based survey of Californians' smoking behavior, conducted in 1996 (8,582 current smokers). Sixty percent of smokers thought the labels light and ultra-light referred to low tar/nicotine cigarettes, or otherwise implied a health claim. This percentage was higher for smokers of low tar/nicotine brands. Among smokers of regular brands, the more highly addicted, those who were trying unsuccessfully to quit, those who had cut consumption or thought about it, and those with health concerns were more likely to have considered switching. While some of these characteristics also were associated with smokers of low tar/nicotine brands, the associations were not as numerous or as strong. We conclude that some smokers appear to view low tar/nicotine brands as one short-term strategy to reduce the harm to their health from smoking without quitting. By implying reduced tar or nicotine exposure, tobacco industry marketing using the labels light and ultra-light is misleading smokers. The use of such labels should be regulated. PMID- 12573177 TI - Smoking reduction practices among African American smokers. AB - Despite smoking fewer cigarettes per day than Caucasians, African Americans bear a disproportionate share of health consequences of smoking. Because the risk of many tobacco-related diseases is dose-dependent, smoking reduction has been suggested as a method to reduce harm for smokers. Little information exists about behavioral smoking-reduction strategies and whether such strategies result in smoking fewer cigarettes. We conducted a survey of 484 African American smokers classified as occasional, light, moderate, and heavy smokers. The survey examined sociodemography, smoking characteristics, and eight smoking reduction strategies, including intentional limiting of smoking, smoking less than half of a cigarette, setting a daily limit for smoking, changing cigarette brand, reducing number of cigarettes, smoking only on some days, switching to a lighter tar cigarette, and not inhaling deeply. Compared to moderate and heavy smokers, occasional and light smokers were more likely to have engaged in most of these strategies. Smokers who used >or= 4 strategies on average smoked 11 cigarettes per day (cpd), compared to 14 cpd and 18 cpd for those who used 1 to 3 strategies and no strategies respectively (p <.0001). After analyses controlled for age, gender, and education, the number of smoking reduction strategies utilized was a significant predictor of smoking 10 or fewer cigarettes per day. This study provides evidence that African American smokers who engaged in multiple smoking reduction strategies smoked fewer cigarettes per day. Smokers not interested in quitting but willing to reduce their smoking should be encouraged to utilize a variety of smoking reduction strategies. PMID- 12573178 TI - Patients' views on smoking cessation and tobacco harm reduction during drug treatment. AB - Efforts to help smokers in drug treatment quit smoking have met with little success. To explore barriers to quitting and interest in tobacco harm reduction we conducted focus groups and interviews among 78 patients from five Methadone Maintenance Treatment (MMT) sites. Measures included a written survey and open ended questions on (a) motivation for quitting, (b) what quit methods worked and what didn't work, and (c) interest in smoking reduction and nicotine maintenance as an alternative to complete cessation. Discussions were audiotaped, transcribed, and coded using computer-based qualitative software; our interobserver reliability was 83%. Successful quitters used in general a combination of quit methods such as prayer, nicotine gum, keeping busy, quitting 'one day at a time,' nicotine patches, deep breathing, and avoidance of triggers. Nicotine craving, rather than withdrawal, was cited as the biggest challenge to staying quit. Some current smokers feared quitting smoking would divert them from their goal of quitting illicit drug use or tapering off MMT. Current smokers were interested in smoking reduction as an alternative to quitting; however, those who had tried but failed to reduce smoking preferred complete cessation. Nicotine maintenance was favored by only a few participants with major health problems who didn't hold out hope for quitting. Findings suggest combination pharmacotherapy could help patients control withdrawal and acute episodes of craving. Patients have a number of skills from coping with illicit drug dependence that are useful in combating nicotine dependence. Behavioral methods and short-term pharmacotherapy to help patients reduce tobacco exposure should be explored. PMID- 12573179 TI - Computer and manual self-help behavioral strategies for smoking reduction: initial feasibility and one-year follow-up. AB - This study sought to test the feasibility of two self-help behavioral interventions to reduce and maintain a 50% reduction in smoking among those unable or unwilling to quit, and to evaluate the impact of smoking reduction on subsequent quit attempts. Ninety-three smokers who desired to reduce rather than quit smoking were entered in the study and randomly assigned to either computerized scheduled gradual reduction (CSGR) or to a manual-based selective elimination reduction (SER). Both groups produced significant reductions in smoking (approximately 10 cigarettes per day, during the 7-week treatment phase), which were maintained over one year. The CSGR group reported greater mean percent reductions in smoking from pre- to post-treatment (37% for CSGR, 20% for SER) and a greater percentage of subjects meeting the 50% reduction goal (30% for CSGR, 16% for SER) compared with the SER group. The groups were comparable, however, on all other outcome measures at post-treatment and at 6- and 12-month follow-up. Although subjects with a current desire for smoking cessation were excluded from this study, one-third of the subjects reported a 24-hour quit attempt in the year following study initiation, and 8.6% of the subjects met 7-day point-prevalence criteria for abstinence (CO validated) at the 12-month follow-up. The results of this study lend support to the feasibility of self-help behavioral interventions to produce sustained reductions in smoking rates without apparent negative impact on subsequent quit attempts. PMID- 12573180 TI - Eclipse can deliver crack cocaine. PMID- 12573181 TI - Treatment options for the twin-twin transfusion syndrome: a review. AB - This article reviews the treatment options of the twin-twin transfusion syndrome (TTTS). No single therapy is associated with a uniformly improved outcome for the involved twins and success is primarily related to gestational age and severity at diagnosis. Treatment options for severe cases include digitalization, ligation of the umbilical cord, serial amniocenteses, septostomy, laser occlusion of placental vessels, and selective feticide. These modalities are associated with significant risks of complications, and variable results of fetal morbidity and mortality. Therefore, they should be considered when risks of withholding treatment clearly outweigh those associated with intervention. PMID- 12573182 TI - Management of twin-twin transfusion syndrome: laying the foundation for future interventional studies. AB - Twin-twin transfusion syndrome is the most common complication of monochorionic pregnancies affecting between 5 and 15% of such pregnancies and accounts for 15 77% of perinatal mortality in twins. The management of twin-twin transfusion syndrome is complex and various treatment modalities have been tried. This review critically evaluates the different management options available for the syndrome and recommends some guidelines for future interventional studies. PMID- 12573183 TI - Conjoined twins in the 16th century. AB - The major morphological types of conjoined twins had all been described by 1600, often in publications that included details of the time and place of birth, morphology, outcome, behaviour, and, on occasion, autopsy findings. These descriptions differ from modern reports in emphasizing the twins' supposed "meaning" rather than their cause. Understanding the symbolic aspects of these accounts is important for their interpretation. This article summarizes the records of 53 cases of conjoined twins born in 16th-century Europe and examines the cultural background of these accounts and the use of the pediatric autopsy as a means of investigating the pathology of twins. Studies of conjoined twins were central to the debate over whether the possession of separate hearts was a prerequisite for individuality. PMID- 12573184 TI - Predicted birthweight for singletons and twins. AB - This population-based study determined the impact of co-twin gender on twin intrauterine growth in addition to their infant gender, maternal height, maternal age and parity on intrauterine growth rate of singletons and twins. All singletons and twins born in Western Australia during the period of 1980 to 1995 were considered for the study. The multiple linear regression models showed that 76% of the variance in the mean birthweight was explained by the selected variables for twins and 51% for singletons. Twins grew more slowly than singletons from 26 weeks gestation. Among twins, opposite-sex twin pairs grew consistently faster than like-sex twins. Primiparous twin pairs grew more slowly than subsequently born twins. These regression equations can be used to assess the appropriateness of intrauterine growth in twin pairs of various gender combinations. PMID- 12573185 TI - Speech in same- and different-sex twins 4 and 5 years old. AB - The purpose of this study was to examine the impact of the sex makeup of pairs of twins on language acquisition. Past research indicated that this variable plays a role in speech problems of twin children. The questions raised were whether being a boy or a girl and having a boy or girl co-twin affected linguistic performance. A language test was given to 30 pairs of boy-girl twins, 16 pairs of boy twins, and 16 pairs of girl twins whose average age was 4 years 8 months. Their test scores confirmed our hypotheses. The poorest performance was obtained by the boy twin pairs and the best performance, by either the girl twin pairs or the different-sex pairs. The results were interpreted in the light of findings on language learning differences between girls and boys, and also in terms of Vygotsky's zone of proximal development. PMID- 12573186 TI - Heritability of educational achievement in 12-year-olds and the overlap with cognitive ability. AB - In order to determine high school entrance level in the Netherlands, nowadays, much value is attached to the results of a national test of educational achievement (CITO), administered around age 12. Surprisingly, up until now, no attention has been paid to the etiology of individual differences in the results of this national test of educational achievement. No attempt has been made to address the question about the nature of a possible association between the results of the CITO and cognitive abilities, as measured by psychometric IQ. The aim of this study is to explore to what extent psychometric IQ and scholastic achievement, as assessed by the CITO high school entrance test, are correlated. In addition, it was investigated whether this expected correlation was due to a common genetic background, shared or nonshared environmental influences common to CITO and intelligence or a combination of these influences. To this end multivariate behavior genetic analyses with CITO and IQ at ages 5, 7, 10 and 12 years have been conducted. The correlations were.41,.50,.60, and.63 between CITO and IQ assessed at age 5, 7, 10, and 12 respectively. The results of the analyses pointed to genetic effects as the main source of variance in CITO and an important source of covariance between CITO and IQ. Additive genetic effects accounted for 60% of the individual differences found in CITO scores in a large sample of Dutch 12-year-olds. This high heritability indicated that the CITO might be a valuable instrument to assess individual differences in cognitive abilities in children but might not be the right instrument to put the effect of education to the test. PMID- 12573187 TI - Variance components models for gene-environment interaction in twin analysis. AB - Gene-environment interaction is likely to be a common and important source of variation for complex behavioral traits. Often conceptualized as the genetic control of sensitivity to the environment, it can be incorporated in variance components twin analyses by partitioning genetic effects into a mean part, which is independent of the environment, and a part that is a linear function of the environment. The model allows for one or more environmental moderator variables (that possibly interact with each other) that may i). be continuous or binary ii). differ between twins within a pair iii). interact with residual environmental as well as genetic effects iv) have nonlinear moderating properties v). show scalar (different magnitudes) or qualitative (different genes) interactions vi). be correlated with genetic effects acting upon the trait, to allow for a test of gene-environment interaction in the presence of gene environment correlation. Aspects and applications of a class of models are explored by simulation, in the context of both individual differences twin analysis and, in a companion paper (Purcell & Sham, 2002) sibpair quantitative trait locus linkage analysis. As well as elucidating environmental pathways, consideration of gene-environment interaction in quantitative and molecular studies will potentially direct and enhance gene-mapping efforts. PMID- 12573188 TI - Variance components models for gene-environment interaction in quantitative trait locus linkage analysis. AB - Gene-environment interaction (G x E) is likely to be a common and important source of variation for complex behavioral traits. Gene-environment interaction, or genetic control of sensitivity to the environment, can be incorporated into variance components twin and sib-pair analyses by partitioning genetic effects into a mean part, which is independent of the environment, and a part that is a linear function of the environment. An approach described in a companion paper (Purcell, 2002) is applied to sib-pair variance components linkage analysis in two ways: allowing for quantitative trait locus by environment interaction and utilizing information on any residual interactions detected prior to analysis. As well as elucidating environmental pathways, consideration of G x E in quantitative and molecular studies will potentially direct and enhance gene mapping efforts. PMID- 12573189 TI - Cardiovascular risk in diabetic women. PMID- 12573190 TI - Clinical trials of vitamin E in coronary artery disease: is it time to reconsider the low-density lipoprotein oxidation hypothesis? AB - A wide range of structurally unrelated antioxidants inhibit atherosclerosis in animal models of hypercholesterolemia, implicating oxidation of low-density lipoprotein (LDL) in the pathogenesis of atherosclerosis. However, most prospective, randomized trials of one proposed antioxidant, vitamin E, have failed to demonstrate any reduction in cardiovascular events in humans with established coronary artery disease. Recent clinical studies suggest that vitamin E is also ineffectual in the primary prevention of atherosclerosis. These observations have led many to question the relevance of LDL oxidation to the pathogenesis of human cardiovascular disease. However, vitamin E's ineffectiveness in clinical trials might result from its failure to act as a physiologically relevant antioxidant. Indeed, vitamin E does not consistently inhibit atherosclerosis in hypercholesterolemic animals, and there is remarkably little evidence that clinically relevant doses of vitamin E result in inhibition of lipid peroxidation in vivo. Collectively, these observations indicate that there is little rationale for using vitamin E to prevent coronary artery disease in humans. They also strongly suggest that it will be critically important to establish that compounds with antioxidant activity in vitro actually prevent oxidative reactions in vivo before embarking on any new clinical trials. PMID- 12573191 TI - Benefits of simvastin in cholesterol lowering. PMID- 12573192 TI - Benefits of atorvastatin in cholesterol lowering. PMID- 12573193 TI - Statins and myotoxicity. AB - The significant age-adjusted decline in cardiovascular mortality that has occurred over the past three decades is multifactorial. However, the advent of statin therapy has markedly facilitated the optimization of dyslipidemia in patients at risk for coronary events. Statin therapy has proven to be effective in reducing morbidity and mortality in large-scale primary and secondary prevention trials. As with all therapies, the administration of 3-hydroxy-3 methylglutaryl coenzyme A (HMG Co A) reductase inhibitors is not without clinical risks. Myopathy, albeit uncommon, was one of the earliest clinical problems associated with statin therapy. Recent data from the large-scale statin mega trials have clarified the quantitative clinical risk-benefit relationship of reductase inhibitors relative to the induction of muscle toxicity. Histopathologic studies have clarified the potential role of statins in the syndrome of myalgias and normal creatine kinase levels. However, the precise mechanism of statin-associated muscle toxicity remains unclear and is potentially related to genetically mediated muscle enzyme defects, drug interactions, intracellular depletion of metabolic intermediates, and intrinsic properties of the statins per se. PMID- 12573194 TI - C-reactive protein: the pawn has been promoted to queen. AB - C-reactive protein (CRP) is one of the acute phase reactants that can increase its serum level up to 100- fold during systemic inflammation. Its clinical use was limited in the past because of its lack of specificity in differentiating infection from other inflammatory processes. With the advent of a high sensitivity assay, CRP was found to be a superb predictor in identifying apparently healthy men and women at risk for developing future cardiovascular events, such as heart attacks and strokes. CRP's predictive power is most likely due to its stability, reproducibility, and proatherogenic properties. Developing consensus to incorporate CRP determination into clinical practice guidelines will be the subject of intense debate and at the same time provide clinical research opportunities in the years to come. PMID- 12573195 TI - Lipoprotein(a) and the atherothrombotic process: mechanistic insights and clinical implications. AB - Although many epidemiologic studies have pointed at an association between plasma levels of lipoprotein(a) (Lp(a)) and cardiovascular risk, the data obtained have been conflicting because of a number of factors, particularly those dealing with plasma storage, lack of assay standardization, population sample size, age, gender, ethnic variations, and variable disease endpoints. Moreover, the attention has been primarily focused on whole Lp(a), with relatively less emphasis on its constituent apolipoprotein(a) and on the apolipoprotein B100 containing lipoprotein, mainly low-density lipoprotein (LDL), to which apolipoprotein(a) is linked. According to recent studies, small-size apolipoprotein(a) isoforms may represent a cardiovascular risk factor either by themselves or synergistically with plasma Lp(a) concentration. Moreover, the density properties of the LDL moiety may have an impact on Lp(a) pathogenicity. It has also become apparent that Lp(a) can be modified by oxidative events and by the action of lipolytic and proteolytic enzymes with the generation of products that exhibit atherothrombogenic potential. The role of the O-glycans linked to the inter-kringle linkers of apolipoprotein(a) is also emerging. This information is raising the awareness of the pleiotropic functions of Lp(a) and is opening new vistas on pathogenetic mechanisms whose knowledge is essential for developing rational therapies against this complex cardiovascular pathogen. PMID- 12573196 TI - Cyclooxygenase-2 inhibitors: is there an association with coronary or renal events? AB - The article is concerned with the effects of specific cyclooxygenase-2 (COX-2) inhibitors and their relationship to thrombotic cardiovascular events and to renal disease. Clinical and experimental aspects of COX-2-specific inhibitors are cited. A COX-2 inhibitor, celecoxib, interferes with myocardial prostacyclin production and also produces hypertension. Data have shown that in animal experiments, celecoxib also lowers myocardial prostaglandin concentration but fails to inhibit thromboxane concentration to the same degree. In the kidney, celecoxib can result in glomerular and interstitial nephritis or papillary necrosis. As in infarcted heart muscle, the COX-2-specific inhibitor celecoxib causes a significant decline in prostaglandin in the renal medulla. It was concluded from both clinical and experimental findings that COX-2 inhibitors can cause thrombotic cardiovascular events as well as renal disease. For these reasons, care should be exercised in administering specific COX-2 inhibitors to patients with pre-existing cardiac or renal disease. PMID- 12573197 TI - Aggressive medical management of coronary artery disease versus mechanical revascularization. AB - The treatment of patients with stable angina has three goals: 1) minimize or eliminate ischemia (silent or symptomatic), 2) reduce morbidity, and 3) decrease mortality. Surgical and now angiographic revascularization procedures are increasingly popular approaches to the management of these patients. The results of randomized, controlled trials suggest that revascularization may not improve survival, but is useful to improve symptoms and exercise capacity. However, these trials are of limited value because they do not reflect current state of the art revascularization techniques or optimal medical management. Because many of these studies were conducted more than a decade ago, patients were recruited before the survival benefits of antiplatelet therapy, b-blockers, angiotensin-converting enzyme inhibitors, and aggressive lipid lowering were accepted. The Clinical Outcomes Utilizing Revascularization and Aggressive Drug Evaluation (COURAGE) trial should help us determine the best approach in these patients. It is a multicenter, randomized trial comparing aggressive medical therapy with aggressive medical therapy with current state of the art percutaneous coronary intervention (PCI) for patients with stable coronary disease. The COURAGE protocol targets global risk reduction emphasizing 1) lifestyle modification, 2) maximal use of drugs to lower blood pressure to Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC) VI goals, 3) maximal use of drugs to lower cholesterol to below National Cholesterol Education Program Adult Treatment Panel III goals for secondary prevention, and 4) maximal use of drug to alleviate anginal symptoms with or without the best interventional devices to conduct PCI. PMID- 12573198 TI - The renin-angiotensin-aldosterone system as a target in coronary disease. AB - The renin-angiotensin-aldosterone system (RAAS) plays a fundamental role in the development of atherosclerosis and adverse cardiovascular events. Traditionally, the pathologic effects of the RAAS were assumed to result from vasoconstriction induced by angiotensin II, and salt and water retention due to aldosterone. However, these hormones also have powerful trophic effects, stimulating increased mass in both the arterial wall and left ventricle. In addition, angiotensin II and aldosterone predispose to vascular inflammation, thrombosis, oxidative stress, and sudden cardiac death. Therapy directed at RAAS overactivity is essential for normalizing the prognosis of most patients with atherosclerosis. An angiotensin-converting enzyme (ACE) inhibitor improves the prognosis of patients with atherosclerosis and/or diabetes even in the setting of normal baseline blood pressure. Angiotensin receptor blocking agents also improve cardiovascular structure and prognosis. Although these agents are better tolerated than ACE inhibitors, they do not appear to be as effective in reducing event rates. Aldosterone receptor blocking agents also improve cardiovascular structure, function, and prognosis. Aldosterone receptor blockers appear to provide additive benefit when used in conjunction with either an ACE inhibitor or an angiotensin receptor blocker. PMID- 12573199 TI - Uncontrolled hypertension as a risk for coronary artery disease: patient characteristics and the role of physician intervention. AB - Hypertension is the most widely treated cardiovascular risk factor, and there is clear evidence of the efficacy of treating systolic and diastolic blood pressure with existing antihypertensive agents in reducing stroke and cardiac disease. However, only about 25% of the US population has blood pressure controlled to at least 140 mm Hg systolic and 90 mm Hg diastolic. Hypertension control is a complex function of patient and physician behavior. Although poor hypertension control has historically been attributed to lack of health insurance or low utilization of available services, recently published analyses of national survey data and local physician and community samples suggest that physicians have a permissive attitude toward isolated mild systolic blood pressure elevations in the range of 140 to 160 mm Hg. The great majority of participants in health surveys report seeing a physician at least two times per year, and several investigators have documented that physicians are unlikely to increase treatment intensity for systolic elevations alone. Physician inaction toward elevated systolic blood pressure may be due to a reluctance to prescribe multiple drugs and/or lack of belief in the benefits of aggressive treatment to lower systolic blood pressure below 140 mm Hg. PMID- 12573200 TI - The role of omega-3 fatty acids in cardiovascular disease. AB - Plant-derived alpha-linolenic acid has been studied in a limited number of investigations. So far, some epidemiologic and a few mechanistic studies suggest a potential of protection from cardiovascular disease, but this potential remains to be proven in intervention studies. In contrast, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are prevalent in fish and fish oils, have been studied in thousands of investigations. A consistent body of evidence has been elaborated in various types of investigations, ultimately demonstrating reduction in total mortality, cardiovascular mortality, and morbidity by ingestion of roughly 1 g/d of EPA plus DHA. Current guidelines, however, do not discern between the omega-3 fatty acids mentioned; in fact, most even do not differentiate polyunsaturated fatty acids at all. Unfortunately, this complicates efficient implementation of an effective means of prophylaxis of atherosclerosis. PMID- 12573201 TI - Myocardial infarction, stroke, and sudden cardiac death may be prevented by influenza vaccination. AB - Acute myocardial infarction (AMI) and atherothrombotic stroke share a common pathogenesis involving disrupted atherosclerotic plaque and intravascular thrombosis. Both AMI and stroke have their peak incidence in winter months. Similarly, the incidence of upper respiratory infections (URIs), 38% of which are due to influenza, also peaks in winter (November and December). URIs result in many biochemical, cellular, and hemostatic changes that could predispose to plaque disruption and thrombosis. Infections, particularly URIs, frequently precede AMI and stroke. Up to 16% of persons older than 60 years of age experience a URI each year. Nineteen percent of those suffering an AMI recall a URI in the 2 weeks prior to their event. Three epidemiologic and one small clinical trial suggest that influenza vaccination is associated with a 50% reduction in incidence of sudden cardiac death, AMI, and ischemic stroke. Influenza vaccine is extremely safe and has a 50% efficacy. Theoretically, up to 104,500 AMIs and 192,000 nonembolic ischemic strokes could be prevented each year by influenza vaccination. PMID- 12573202 TI - Coronary arterial remodeling: from bench to bedside. AB - Coronary arterial remodeling describes changes of vessel size at the site of atherosclerotic lesions. Positive remodeling (expansion) of early lesions maintains lumen size despite plaque accumulation. In contrast, negative remodeling (shrinkage) contributes to luminal stenosis independent of plaque accumulation. Because of these adaptive changes, plaque progression/regression is not closely reflected in luminal size. Histologic studies have demonstrated that the pathophysiologic role of arterial remodeling is more complex than a mere compensatory process. Surprisingly, there is a consistent association between positive arterial remodeling, local inflammatory response, and plaque vulnerability. In vivo tomographic imaging techniques, in particular intravascular ultrasound, and potentially computed tomography and magnetic resonance imaging, allow the observation of remodeling in clinical settings. The integration of basic knowledge about arterial remodeling with clinical observations from in vivo imaging could lead to a better understanding of plaque progression, regression, and vulnerability and may eventually have implications for disease prevention. PMID- 12573203 TI - Transgenics: are these foods beneficial or harmful? PMID- 12573204 TI - Use of OM-85 BV in children suffering from recurrent respiratory tract infections and subnormal IgG subclass levels. AB - BACKGROUND: Recurrent acute respiratory tract infections (RARTIs) in children are related to IgG subclass deficiencies. The aim of the trial was to evaluate the effect of OM-85 BV in the number of RARTIs as well as in the IgG subclass levels. METHODS: This was a randomized, double-blind, placebo-controlled clinical trial. Patients of ages three to six years, having three or more documented ARTIs during the last six months with subnormal IgG subclass levels were included. Patients took either one capsule of OM-85 BV (3.5 mg) or placebo orally every day for ten consecutive days per month during three consecutive months. Patients were followed three further months without drug intake. IgG subclass levels were determined before and after treatment. RESULTS: IgG4 levels diminished after the OM-85 BV treatment (-3 [-8.0, -1.0] median difference [95 % CI] p < 0.05 by Wilcoxon test). No other significant changes in IgG subclasses were observed. After six months the patients in the OM-85 BV group (n = 20) experienced 2.8 1.4 (mean SD) ARTIs, while the patients in the placebo group (n = 20) suffered 5.2 1.5 ARTIs (-2.4 [3.3, -1.5] mean difference [95 % CI] p < 0.001 by Student's t test). Three patients with OM-85 BV had gastrointestinal events related to drug administration, as well as three placebo patients. CONCLUSION: This study demonstrated the clinical benefit of OM-85 BV in patients suffering from RARTIs and subnormal levels of IgG subclasses. This trial opens new perspectives in the research of the mechanism of action of OM-85 BV. PMID- 12573205 TI - The association between delayed type hypersensitivity reaction to Mycobacterium tuberculosis and atopy in asthmatic children. AB - BACKGROUND: It has been postulated that there is an inverse association between mycobacterium tuberculosis infection and atopy. We aimed to investigate if there is a similar relation in our study group, consisting 252 asthmatic children. METHODS: In tuberculin testing indurations greater than or equal to 5 mm were accepted as positive. The most common aeroallergens were used in skin prick testing and reactions > or = 3 mm were accepted as positive. RESULTS: In 139 patients PPD was negative, where as in 113 patients PPD was positive. Among the PPD (-) patients skin prick test was positive in 64 % (n = 89). Among the PPD (+) patients skin prick test was positive in 71 % (n = 80). As the two groups were compared for having positive skin prick test reactions no statistically significant difference was detected between them (p = 0.283). CONCLUSIONS: Tuberculin reactivity is not inversely associated with atopy in asthmatic children. PMID- 12573206 TI - [The levels of IgG subclasses in respiratory allergic pathology in the childhood]. AB - OBJECTIVES: To assess the existence of immunologic alterations, referring to IgG subclasses, in the respiratory allergic pathology in the childhood. MATERIAL AND METHODS: This is a prospective study that compares the IgG subclasses levels in a sample of patient with allergic breathing illness (n = 169) and a group control (n = 130) with ages range from one month to 13 years old. The statistical analysis includes a descriptive statistic and a comparative statistic carrying out comparison of means between both groups with the Welch test and the Student's T test. RESULTS: The mean rate of IgG1 in the group with breathing allergic pathology being of 578,1 mg/dl and in the control group, being of 632,78 mg/dl. The mean rate of IgG2 in the group with breathing allergic pathology being of 106,12 mg/dl and in the control group, being of 142,38 mg/dl. The mean rate of IgG3 in the group with breathing allergic pathology being of 53,73 mg/dl and in the control group, being of 63,78 mg/dl. The children with breathing allergic pathology have significantly decreased IgG1, IgG2, IgG3 levels in comparison to the control group (p < 0.001). The mean rate of IgG4 in the group with breathing allergic pathology being of 25,86 mg/dl and in the control group, being of 13,89 mg/dl. The children with breathing allergic pathology have significantly elevated IgG4 levels in comparison to the control group (p < 0.001). CONCLUSIONS: The findings obtained suggest that an relation exists in the breathing allergic processes with the IgG subclasses levels disturbances and also that there is an alteration of the immune response maturation in relation to subclasses IgG1, IgG2, IgG3. The valuation of the levels of these immunoglobulins can be useful in the pathogenic diagnosis of the allergic processes. PMID- 12573207 TI - A pre-seasonal birch/hazel sublingual immunotherapy can improve the outcome of grass pollen injective treatment in bisensitized individuals. A case-referent, two-year controlled study. AB - BACKGROUND: The study tests the hypothesis of a reduction of priming due to tree allergy in patients sensitised to both birch/hazel and grass pollen undergoing an associated preseasonal Sublingual/Injective immunotherapy. METHODS: 36 out of 49 bisensitized candidates were pair-matched into 18 case-referent couples. During two years all patients were administered preseasonal grass-SIT and one patient in each couple received also birch/hazel-SLIT. Diary cards were fulfilled for three consecutive grass pollen seasons. Specific Nasal Provocation Test (NPT) for grass and aspecific bronchial challenge were done; sera were analyzed for specific IgE and IgG. RESULTS: During the peak of the grass pollen season both groups showed a significant improvement in total symptom-score. Conjunctivitis and cough improved significantly more in patients with associated therapies. While antihistamine score decreased significantly in both groups, antiasthmatics did only in the SLIT SIT group. The follow-up documented a significant increase in grass- and birch specific IgG and a decrease in grass-specific IgE. Grass-NPT threshold was clearly higher in SLIT-SIT-group (p = 0.01) and only in this group PD20 methacholine improved significantly (p < 0.05). CONCLUSIONS: Combined birch/hazel SLIT and grass-SIT are safe and improve clinical outcomes of SIT alone in young bisensitized patients. Priming reduction is supported by specific NPT and bronchial hyperresponsiveness. PMID- 12573208 TI - Enhancement of tomato allergenicity after treatment with plant hormones. AB - BACKGROUND: Practical applications to enhance the productivity of agriculture by using plants with improved resistance to pathogens are expected to increase in the near future. Although tomato has been widely investigated for breeding purposes, there have been no studies on tomato allergenicity after plant hormones treatments. METHODS: Prick by prick tests were carried out with different tomato samples (fruits grown under biological conditions without addition of chemical products, and treated with ethylene and salicylic acid) in eight patients with ages between 12 and 27 years who suffered from anaphylaxis episodes after eating raw tomatoes. An immunoblot experiment with the different tomato extracts was performed using sera from these eight patients and controls. RESULTS: The wheals obtained in prick tests were significantly higher with the extracts of tomato treated with ethylene and SAA (chi(2) = 31.3, p < 0.0001) and the patients who presented higher wheal diameters in skin tests were those who had more severe episodes of anaphylaxis. Neither the protein stain nor the IgE immunodetection patterns clearly varied between the untreated and the hormone-treated samples. CONCLUSIONS: In the case of anaphylaxis induced by tomato, the treatment with plant hormones induced a higher cutaneous response than with non-treated tomato, but the "in vitro" response was similar. PMID- 12573209 TI - Anaphylaxis to Linum. AB - BACKGROUND: Flax (Linum usitatissimum) seeds are increasingly used in bread and as laxatives. Hypersensitivity to linseeds has been infrequently described, and we report a case of anaphylaxis induced by linseed ingestion in a 39-year-old woman. METHODS AND RESULTS: The clinical course, as well as positive skin prick tests and histamine release tests performed with linseed extracts, suggested a type I hypersensitivity as the underlying cause for the patient's multisystemic involvement. The presence of linum-specific IgE in her serum was confirmed by immunoCAP assay. CONCLUSION: Linum seeds might be a source of allergic sensitization that should be taken into account due to its widespread distribution at health food stores. PMID- 12573210 TI - [Angioedema due to sensitization to chicken meat]. AB - BACKGROUND: Egg is the most frequent cause of food allergy in children. The bird egg syndrome, found in a group of patients sensitized to egg through bird proteins, was infrequent in children. We report a patient with former history of hypersensitivity to egg who developed episodes of angioedema after ingestion of hen meat. METHODS: Prick testing with egg and their different antigenic protein fractions, alpha-livetin and chicken meat was performed. Antigens of hen meat were used for the skin prick test and prick-by-prick. Serum-specific IgE was identified with use of the CAP techniques and SDS-PAGE Immunoblotting. RESULTS: Prick test was positive with egg yolk, alpha-livetin and chicken meat. A prick-by prick test with hen meat resulted positive in our patient, but the same test in four controls patients were negative. Serum specific IgE was positive for egg yolk and hen meat. CONCLUSION: Allergy reactions to hen meat are exceptional. We report a case of children with allergy to egg proteins and hen meat that suggest an IgE mediated hypersensitivity reaction. Skin test reveal sensitivity to egg yolk and alpha-livetin, but this pattern of sensitization was infrequent in children. PMID- 12573211 TI - Non-pigmenting cutaneous-mucosal fixed drug eruption due to piroxicam. AB - BACKGROUND: Piroxicam is a widely used anti-inflammatory drug. Most adverse reactions affect gastrointestinal system, liver and skin. Fixed drug eruption although very unusual, has also been described, but with cutaneous involvement exclusively. We present the case of a 49-year-old man who suffered three episodes of fixed drug eruption with cutaneous-mucosal involvement, even simulating an autoimmune disease, whenever he was treated with oral piroxicam. METHODS AND RESULTS: He was patch tested on normal skin with the GEIDC standard series and an NSAIDs series. He was patch tested on normal skin and on fixed eruption with piroxicam, meloxicam and tenoxicam (all of them 1 % pet). Oral challenge test was not performed due to the severity and reproducibility in previous reactions. Results showed a positive patch test to piroxicam (1 % pet) on fixed eruption, with negative results to the rest. CONCLUSIONS: Adverse drug reactions may present a wide variability of clinical symptoms. In these situations an accurate clinical history is necessary. To our knowledge this is the 1st report of non pigmenting fixed drug eruption with cutaneous-mucosal involvement due to piroxicam. Cross-reactivity between oxicams could not be demonstrated by patch test on fixed eruption. PMID- 12573212 TI - Autoimmune hepatitis and odour of fish food proteins. PMID- 12573213 TI - POLTERGEIST encodes a protein phosphatase 2C that regulates CLAVATA pathways controlling stem cell identity at Arabidopsis shoot and flower meristems. AB - BACKGROUND: Receptor kinases are a large gene family in plants and have more than 600 members in Arabidopsis. Receptor kinases in plants regulate a broad range of developmental processes, including steroid hormone perception, organ elongation, self-incompatibility, and abscission. Intracellular signaling components for receptor kinases in plants are largely unknown. The CLAVATA 1 (CLV1) receptor kinase in Arabidopsis regulates stem cell identity and differentiation through its repression of WUSCHEL (WUS) expression. Mutations at the POLTERGEIST (POL) gene were previously described as phenotypic suppressors of mutations within the CLV1 gene. Genetic evidence placed POL as a downstream regulator of CLAVATA1 signaling. RESULTS: We provide evidence that POL functions in both the CLV1-WUS pathway and a novel WUS-independent CLV1 pathway regulating stem cell identity. We demonstrate that POL encodes a protein phosphatase 2C (PP2C) with a predicted nuclear localization sequence, indicating that it has a role in signal transduction downstream of the CLV1 receptor. The N terminus of POL has a possible regulatory function, and the C terminus has PP2C-like phosphatase catalytic activity. Although the POL catalytic domain is conserved in other PP2Cs, the POL protein represents a unique subclass of plant PP2Cs. POL is broadly expressed throughout the plant. CONCLUSIONS: POL represents a novel component of the CLV1 receptor kinase signaling pathway. The ubiquitous expression of POL and pol phenotypes outside the meristem suggest that POL may be a common regulator of many signaling pathways. PMID- 12573214 TI - Nocturnin, a deadenylase in Xenopus laevis retina: a mechanism for posttranscriptional control of circadian-related mRNA. AB - BACKGROUND: Different types of regulation are utilized to produce a robust circadian clock, including regulation at the transcriptional, posttranscriptional, and translational levels. A screen for rhythmic messages that may be involved in such circadian control identified nocturnin, a novel gene that displays high-amplitude circadian expression in the Xenopus laevis retina, with peak mRNA levels in the early night. Expression of nocturnin mRNA is confined to the clock-containing photoreceptor cell layer within the retina. RESULTS: In these studies, we show that nocturnin removes the poly(A) tail from a synthetic RNA substrate in a process known as deadenylation. Nocturnin nuclease activity is magnesium dependent, as the addition of EDTA or mutation of the residue predicted to bind magnesium disrupts deadenylation. Substrate preference studies show that nocturnin is an exonuclease that specifically degrades the 3' poly(A) tail. While nocturnin is rhythmically expressed in the cytoplasm of the retinal photoreceptor cells, the only other described vertebrate deadenylase, PARN, is constitutively present in most retinal cells, including the photoreceptors. CONCLUSIONS: The distinct spatial and temporal expression of nocturnin and PARN suggests that there may be specific mRNA targets of each deadenylase. Since deadenylation regulates mRNA decay and/or translational silencing, we propose that nocturnin deadenylates clock-related transcripts in a novel mechanism for posttranscriptional regulation in the circadian clock or its outputs. PMID- 12573215 TI - Real-time visualization of ZBP1 association with beta-actin mRNA during transcription and localization. AB - BACKGROUND: mRNA localization in somatic cells is an important mechanism for gene expression regulation. In fibroblasts, the protein ZBP1 associates with the sequence that localizes beta-actin mRNA to the leading edge of fibroblasts, augmenting motility. beta-actin mRNA localizes in a cytoskeleton-dependent manner, depending on intact actin and myosin ATP-hydrolysis, and is largely bound to the actin cytoskeleton. The ZBP1 protein contains four KH RNA binding domains and a classic RBD RNA binding domain. It also contains a putative nuclear import and export sequence, suggesting a nuclear phase in this protein's function. RESULTS: Using high-speed imaging, we show here the targeting of this RNA binding protein to beta-actin pre-mRNA transcripts in the nuclei of living cells and measure the residence time of the RNA-protein complex before it leaves the transcription site. Then, the RNA-protein particle is exported to the cytoplasm, where it localizes at velocities of 0.6 microm/s by using actin filaments and/or microtubules. This RNA-ZBP1 complex is required for cytoplasmic localization in fibroblasts; mislocalizing the protein also mislocalizes the RNA, and expressing the protein in a ZBP1-deficient cell line induces beta-actin mRNA localization. CONCLUSIONS: This work demonstrates that the RNA-protein association, essential for cytoplasmic localization, begins as soon as the RNA is transcribed. The ZBP1 then forms a ribonucleoprotein particle and moves in a myosin-dependent fashion by using the cytoskeleton for directional transport. PMID- 12573216 TI - Depletion of Drad21/Scc1 in Drosophila cells leads to instability of the cohesin complex and disruption of mitotic progression. AB - BACKGROUND: The coordination of cell cycle events is necessary to ensure the proper duplication and dissemination of the genome. In this study, we examine the consequences of depleting Drad21 and SA, two non-SMC subunits of the cohesin complex, by dsRNA-mediated interference in Drosophila cultured cells. RESULTS: We have shown that a bona fide cohesin complex exists in Drosophila embryos. Strikingly, the Drad21/Scc1 and SA/Scc3 non-SMC subunits associate more intimately with one another than they do with the SMCs. We have observed defects in mitotic progression in cells from which Drad21 has been depleted: cells delay in prometaphase with normally condensed, but prematurely separated, sister chromatids and with abnormal spindle morphology. Much milder defects are observed when SA is depleted from cells. The dynamics of the chromosome passenger protein, INCENP, are affected after Drad21 depletion. We have also made the surprising observation that SA is unstable in the absence of Drad21; however, we have shown that the converse is not true. Interference with Drad21 in living Drosophila embryos also has deleterious effects on mitotic progression. CONCLUSIONS: We conclude that Drad21, as a member of a cohesin complex, is required in Drosophila cultured cells and embryos for proper mitotic progression. The protein is required in cultured cells for chromosome cohesion, spindle morphology, dynamics of a chromosome passenger protein, and stability of the cohesin complex, but apparently not for normal chromosome condensation. The observation of SA instability in the absence of Drad21 implies that the expression of cohesin subunits and assembly of the cohesin complex will be tightly regulated. PMID- 12573217 TI - Stress grows wings: environmental induction of winged dispersal males in Cardiocondyla ants. AB - Dispersal is advantageous, but, at the same time, it implies high costs and risks. Due to these counteracting selection pressures, many species evolved dispersal polymorphisms, which, in ants, are typically restricted to the female sex (queens). Male polymorphism is presently only known from a few genera, such as Cardiocondyla, in which winged dispersing males coexist with wingless fighter males that mate exclusively inside their maternal nests. We studied the developmental mechanisms underlying these alternative male morphs and found that, first, male dimorphism is not genetically determined, but is induced by environmental conditions (decreasing temperature and density). Second, male morph is not yet fixed at the egg stage, but it differentiates during larval development. This flexible developmental pattern of male morphs allows Cardiocondyla ant colonies to react quickly to changes in their environment. Under good conditions, they invest exclusively in philopatric wingless males. But, when environmental conditions turn bad, colonies start to produce winged dispersal males, even though these males require a many times higher investment by the colony than their much smaller wingless counterparts. Cardiocondyla ants share this potential of optimal resource allocation with other colonial animals and some seed dimorphic plants. PMID- 12573219 TI - Genes encoding A-type flavoproteins are essential for photoreduction of O2 in cyanobacteria. AB - O(2) photoreduction by photosynthetic electron transfer, the Mehler reaction, was observed in all groups of oxygenic photosynthetic organisms, but the electron transport chain mediating this reaction remains unidentified. We provide the first evidence for the involvement of A-type flavoproteins that reduce O(2) directly to water in vitro. Synechocystis sp. strain PCC 6803 mutants defective in flv1 and flv3, encoding A-type flavoproteins, failed to exhibit O(2) photoreduction but performed normal photosynthesis and respiration. We show that the light-enhanced O(2) uptake was not due to respiration or photorespiration. After dark acclimation, photooxidation of P(700) was severely depressed in mutants Deltaflv1 and Deltaflv3 but recovered after light activation of CO(2) fixation, which gives P(700) an additional electron acceptor. Inhibition of CO(2) fixation prevented recovery but scarcely affected P(700) oxidation in the wild type, where the Mehler reaction provides an alternative route for electrons. We conclude that the source of electrons for O(2) photoreduction is PSI and that the highly conserved A-type flavoproteins Flv1 and Flv3 are essential for this process in vivo. We propose that in cyanobacteria, contrary to eukaryotes, the Mehler reaction produces no reactive oxygen species and may be evolutionarily related to the response of anaerobic bacteria to O(2). PMID- 12573218 TI - Nanos plays a conserved role in axial patterning outside of the Diptera. AB - Axial patterning is a fundamental event in early development, and molecules involved in determining the body axes provide a coordinate system for subsequent patterning. While orthologs of Drosophila bicoid and nanos play a conserved role in anteroposterior (AP) patterning within at least a subset of Diptera, conservation of this process has not yet been demonstrated outside of the flies. Indeed, it has been argued that bicoid, an instrumental "anterior" factor in Drosophila melanogaster, acquired this role during the evolution of more-derived dipterans. Interestingly, the interaction of Drosophila maternal nanos and maternal hunchback provides a system for patterning the AP axis that is partially redundant to the anterior system. Previous studies in grasshoppers suggest that hunchback may play a conserved role in axial patterning in this insect, but this function may be supplied solely by the zygotic component of hunchback expression. Here we provide evidence that the early pattern of zygotic grasshopper Hunchback expression is achieved through translational repression that may be mediated through the action of grasshopper nanos. This is consistent with the notion that an anterior gradient system is not necessary in all insects and that the posterior pole "probably conveys longitudinal polarity on the ensuing germ anlage". PMID- 12573220 TI - Posttranscriptional gene silencing is not compromised in the Arabidopsis CARPEL FACTORY (DICER-LIKE1) mutant, a homolog of Dicer-1 from Drosophila. AB - Posttranscriptional silencing (PTGS) in plants, nematodes, Drosophila, and perhaps all eukaryotes operates by sequence-specific degradation or translational inhibition of the target mRNA. These processes are mediated by duplexed RNA. In Drosophila and nematodes, double-stranded (ds)RNA or self-complementary RNA is processed into fragments of approximately 21 nt by Dicer-1. These small interfering RNAs (siRNAs) serve as guides to target degradation of homologous single-stranded (ss)RNA. In some cases, the approximately 21 nt guide fragments derived from endogenous, imperfectly self-complementary RNAs cause translational inhibition of their target mRNAs, with which they have substantial, but not perfect sequence complementarity. These small temporal RNAs (stRNAs) belong to a class of noncoding microRNAs (miRNAs), 20-24 nt in length, that are found in flies, plants, nematodes, and mammals. In nematodes, the Dicer-1 enzyme catalyzes the production of both siRNA and stRNA. Mutation of the Arabidopsis Dicer-1 homolog, CARPEL FACTORY (CAF), blocks miRNA production. Here, we report that the same caf mutant does not block either PTGS or siRNA production induced by self complementary hairpin RNA. This suggests either that this mutation only impairs miRNA formation or, more interestingly, that plants have two distinct dicer-like enzymes, one for miRNA and another for siRNAi production. PMID- 12573221 TI - Sequential recruitment of HAT and SWI/SNF components to condensed chromatin by VP16. AB - Eukaryotic transcription initiation requires the complex dynamics of hundreds of proteins, many of which are found in large multisubunit complexes. Recent experiments have suggested stepwise recruitment of preassembled complexes, including chromatin remodeling, general transcription factor, mediator, and polymerase complexes, in which the actual order of recruitment may vary for different promoters. How do these complexes access target sequences contained within tightly condensed chromatin? While chromatin remodeling activities may facilitate the accessibility of large transcription and polymerase complexes to promoters, it is not known how they themselves are targeted within condensed chromatin. Gene activation in the context of condensed chromatin does occur. A yeast acidic activator, Gal4, can overcome heterochromatin gene silencing in Drosophila, and the addition of LCRs (locus control regions) to transgenes overcomes position effect silencing, even within centromeric chromatin. Here, we directly visualize the recruitment of HAT and SWI/SNF components after tethering the VP16 acidic activation domain within condensed chromatin. A recruitment delay of tens to hundreds of minutes for catalytic HAT subunits and SWI/SNF subunits, relative to other HAT and SWI/SNF components, suggests sequential recruitment/assembly of chromatin remodeling complexes within condensed chromatin. PMID- 12573222 TI - NAADP: a new second messenger for glucose-induced Ca2+ responses in clonal pancreatic beta cells. AB - Important questions remain concerning how elevated blood glucose levels are coupled to insulin secretion from pancreatic beta cells and how this process is impaired in type 2 diabetes. Glucose uptake and metabolism in beta cells cause the intracellular Ca(2+) concentration ([Ca(2+)](i)) to increase to a degree necessary and sufficient for triggering insulin release. Although both Ca(2+) influx and Ca(2+) release from internal stores are critical, the roles of inositol 1,4,5-trisphosphate (IP(3)) and cyclic adenosine dinucleotide phosphate ribose (cADPR) in regulating the latter have proven equivocal. Here we show that glucose also increases [Ca(2+)](i) via the novel Ca(2+)-mobilizing agent nicotinic acid adenine dinucleotide phosphate (NAADP) in the insulin-secreting beta-cell line MIN6. NAADP binds to specific, high-affinity membrane binding sites and at low concentrations elicits robust Ca(2+) responses in intact cells. Higher concentrations of NAADP inactivate NAADP receptors and attenuate the glucose-induced Ca(2+) increases. Importantly, glucose stimulation increases endogenous NAADP levels, providing strong evidence for recruitment of this pathway. In conclusion, our results support a model in which NAADP mediates glucose-induced Ca(2+) signaling in pancreatic beta cells and are the first demonstration in mammalian cells of the presence of endogenous NAADP levels that can be regulated by a physiological stimulus. PMID- 12573223 TI - ADF/cofilin controls cell polarity during fibroblast migration. AB - To migrate, normally a cell must establish morphological polarity and continuously protrude a single lamellipodium, polarized in the direction of migration. We have previously shown that actin filament disassembly is necessary for protrusion of the lamellipodium during fibroblast migration. As ADF/cofilin (AC) proteins are essential for the catalysis of filament disassembly in cells, we assessed their role in polarized lamellipodium protrusion in migrating fibroblasts. We compared the spatial distribution of AC and the inactive, phosphorylated AC (pAC) in migrating cells. AC, but not pAC, localized to the lamellipodium. To investigate a role for AC in cell polarity, we increased the proportion of pAC in migrating fibroblasts by overexpressing constitutively active (CA) LIM kinase 1. In 87% of cells expressing CA LIM kinase, cell polarity was abolished. In such cells, the single polarized lamellipodium was replaced by multiple nonpolarized lamellipodia, which, in contrast to nonexpressing migrating cells, stained for pAC. Cell polarity was rescued by coexpressing an active, nonphosphorylatable Xenopus AC (CA XAC) with the CA LIMK. Furthermore, overexpressing a pseudophosphorylated (less active) XAC by itself also abolished cell polarity. We conclude that locally maintaining ADF/cofilin in the active, nonphosphorylated state within the lamellipodium is necessary to maintain polarized protrusion during cell migration. PMID- 12573224 TI - Ubiquitin signals protein trafficking via interaction with a novel ubiquitin binding domain in the membrane fusion regulator, Vps9p. AB - The conserved vacuolar protein-sorting (Vps) pathway controls the trafficking of proteins to the vacuole/lysosome. Both the internalization of ubiquitylated cargo from the plasma membrane and its sorting at the late endosome via the Vps pathway depend on ubiquitin (Ub) binding motifs present in trafficking regulators. Here we report that Ub controls yet a third step in the Vps pathway. Vps9p, which promotes endosomal and Golgi-derived vesicle fusion, binds directly to Ub via a Cue1p-homologous (CUE) domain. The CUE domain is structurally related to the Ub associated (UBA) domain. In an assay for vacuolar delivery of a transmembrane receptor fused to Ub, a Ub mutation impairing interaction with Vps9p led to a cytoplasmic block in receptor trafficking. This block resembled that of a receptor fused to wild-type Ub but expressed in a vps9-null background. Strikingly, this trafficking defect caused by a mutant Ub was rescued by deletion of the Vps9p CUE domain, indicating that lack of the CUE domain renders Vps9p independent of Ub for activation in vivo. We thus provide evidence for biochemical and genetic interactions between Ub and a novel Ub binding domain in Vps9p. Ub plays a positive role, whereas the CUE domain plays both positive and negative roles in Vps9p function in trafficking. PMID- 12573226 TI - Push and pull on the new "C" word. PMID- 12573225 TI - Human cloning spectre rises again. PMID- 12573228 TI - Polycomb group proteins. PMID- 12573229 TI - How does the beetle cross the road? PMID- 12573230 TI - Q & A. PMID- 12573231 TI - Colour vision: colouring the dark. AB - Humans lose colour vision at night and it has often been assumed that this happens to other animals as well. It is not true of nocturnal moths, however: a recent study has shown that the elephant hawk moth makes use of trichromatic colour vision when seeking flowers by starlight. PMID- 12573232 TI - Human prehistory: the message from linkage disequilibrium. AB - The vast amount of information being gathered about human DNA sequence variation raises the question of what these data can tell us about events in our past. A new way has been found by which patterns of linkage disequilibrium can be used to detect the effects of natural selection in human prehistory. PMID- 12573233 TI - Centrosomes: coiled-coils organize the cell center. AB - The centrosome serves as a structural context for cytoplasmic organization. Recent studies on mutants of the nematode worm Caenorhabditis elegans have provided new insight into the framework to which microtubules and key regulators of centrosome behavior are anchored. PMID- 12573234 TI - Auditory neuroscience: the salience of looming sounds. AB - Sounds that move towards us have a greater biological salience than those that move away. Recent studies in human and non-human primates have demonstrated a perceptual and behavioural priority for such looming sounds that is also reflected in an asymmetric pattern of cortical activation. PMID- 12573235 TI - Cell adhesion: a FERM grasp of membrane dynamics. AB - Two recent papers provide the first evidence for a common mechanism of targeting and activation of an enzyme that is important in the rapid regulation of both focal adhesion assembly during cell migration and synaptic vesicle recycling at nerve terminals. PMID- 12573236 TI - Organogenesis: making pancreas from liver. AB - Making endocrine pancreas cells at will is one of the major goals of cellular based therapies for diabetes. The experimentally induced conversion of hepatocytes into pancreatic cells, using a modified version of the transcription factor Pdx-1, may provide an alternative to stem cell approaches. PMID- 12573237 TI - Adenovirus infection: taking the back roads to viral entry. AB - Certain virus receptors are sequestered on the basolateral surface of polarized epithelial cells. A recent study has shown how adenovirus--and perhaps other viruses--are able to overcome this physical barrier. PMID- 12573238 TI - Sex determination: sex on the brain? AB - Five newly identified sex-biased transcripts in Drosophila are full of surprises: although they are found in fly heads, and manipulating two of them affects mating behaviors, their genes are expressed sex-specifically in non-neural tissues. The way these genes are regulated suggests new complexities in the sex determination pathway. PMID- 12573239 TI - Chromosome cohesion and separation: from men and molecules. AB - Sister chromatid cohesion and separation are fundamental for accurate genome inheritance over cell generations. Work over recent years has established the existence of a chromosomal protein complex, cohesin, that connects sister chromatids from the time they are generated in S phase onwards, and which is destroyed at the onset of anaphase through cleavage by the protease separase. Over the last year, the function of cohesin has been investigated in higher eukaryotes, including humans, with results that have uncovered important new aspects of this process. The first structural views of cohesin have become available, and significant steps been made towards a mechanistic understanding of chromosome cohesion. Studies on separase have revealed new levels of regulation of chromosome segregation. PMID- 12573240 TI - Crystal structure of human phosphoglucose isomerase and analysis of the initial catalytic steps. AB - The second enzyme in the glycolytic pathway, phosphoglucose isomerase (PGI), catalyses an intracellular aldose-ketose isomerization. Here we describe the human recombinant PGI structure (hPGI) solved in the absence of active site ligands. Crystals isomorphous to those previously reported were used to collect a 94% complete data set to a limiting resolution of 2.1 A. From the comparison between the free active site hPGI structure and the available human and rabbit PGI (rPGI) structures, a mechanism for protein initial catalytic steps is proposed. Binding of the phosphate moiety of the substrate to two distinct elements of the active site is responsible for driving a series of structural changes resulting in the polarisation of the active site histidine, priming it for the initial ring-opening step of catalysis. PMID- 12573241 TI - Identification of phosphorylation sites within the SH3 domains of Tec family tyrosine kinases. AB - Tec family protein tyrosine kinases (TFKs) play a central role in hematopoietic cellular signaling. Initial activation takes place through specific tyrosine phosphorylation situated in the activation loop. Further activation occurs within the SH3 domain via a transphosphorylation mechanism, which for Bruton's tyrosine kinase (Btk) affects tyrosine 223. We found that TFKs phosphorylate preferentially their own SH3 domains, but differentially phosphorylate other member family SH3 domains, whereas non-related SH3 domains are not phosphorylated. We demonstrate that SH3 domains are good and reliable substrates. We observe that transphosphorylation is selective not only for SH3 domains, but also for dual SH3SH2 domains. However, the dual domain is phosphorylated more effectively. The major phosphorylation sites were identified as conserved tyrosines, for Itk Y180 and for Bmx Y215, both sites being homologous to the Y223 site in Btk. There is, however, one exception because the Tec-SH3 domain is phosphorylated at a non-homologous site, nevertheless a conserved tyrosine, Y206. Consistent with these findings, the 3D structures for SH3 domains point out that these phosphorylated tyrosines are located on the ligand-binding surface. Because a number of Tec family kinases are coexpressed in cells, it is possible that they could regulate the activity of each other through transphosphorylation. PMID- 12573242 TI - Cloning and expression of the gene encoding catalytic subunit of thermostable glucose dehydrogenase from Burkholderia cepacia in Escherichia coli. AB - We have cloned a 1620-nucleotide gene encoding the catalytic subunit (alpha subunit) of a thermostable glucose dehydrogenase (GDH) from Burkholderia cepacia. The FAD binding motif was found in the N-terminal region of the alpha subunit. The deduced primary structure of the alpha subunit showed about 48% identity to the catalytic subunits of sorbitol dehydrogenase (SDH) from Gluconobacter oxydans and 2-keto-D-gluconate dehydrogenases (2KGDH) from Erwinia herbicola and Pantoea citrea. The alpha subunit of B. cepacia was expressed in Escherichia coli in its active water-soluble form, showing maximum dye-mediated GDH activity at 70 degrees C, retaining high thermal stability. A putative open reading frame (ORF) of 507 nucleotides was also found upstream of the alpha subunit encoding an 18 kDa peptide, designated as gamma subunit. The deduced primary structure of gamma subunit showed about 30% identity to the small subunits of the SDH from G. oxydans and 2KGDHs from E. herbicola and P. citrea. PMID- 12573243 TI - Construction and characterization of a chimeric myoglobin. AB - In order to investigate the functional and structural role of modular structure in globins, we have engineered a chimeric myoglobin (ChimMb) in which the first and third exon come from the gene coding for the sperm whale Mb and the second exon from the gene coding for Aplysia limacina Mb. This ChimMb, fused to the Maltose Binding Protein (MBP) and expressed in Escherichia coli as an apoprotein, binds protoheme in a 1:1 stoichiometric ratio. Based on some functional and spectroscopic properties, we conclude that the central core of the ChimMb (which derives from A. limacina) is native-like. On the other hand, the ChimMb deprived (by proteolytic digestion) of the fused MBP displays a considerably reduced stability. These results suggest that the sperm whale A-G-H nucleus does not contribute significantly to the overall stability of the ChimMb. PMID- 12573245 TI - Sequence determination of three cuticular proteins and isoforms from the migratory locust, Locusta migratoria, using a combination of Edman degradation and mass spectrometric techniques. AB - The cuticle (exoskeleton) is a characteristic structure of insects and other arthropods. It is an extracellular layer which surrounds and protects the insect, and it is composed of proteins, lipids, water molecules, phenolic materials and chitin. Four proteins isolated from the thorax and femur cuticle of pharate adult migratory locust, Locusta migratoria, have been purified by ion-exchange chromatography and reversed-phase high performance liquid chromatography (RP HPLC). Their amino acid sequences were determined by combined use of mass spectrometry and automated Edman degradation. The cuticular extract was also separated by two-dimensional gel electrophoresis. In order to localize and identify the position of the proteins in the gel, a number of gel spots were excised and the proteins electroeluted. The molecular mass of some of the electroeluted proteins was determined by means of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) as well as by electrospray mass spectrometry (ESI-MS). Two of the sequenced proteins exist as pairs of closely related isoforms; one of the pairs contains the conserved 68-residue RR-2 motif, common for proteins from solid cuticles, and the other proteins contain the short motif Ala-Ala-Pro-Ala/Val repeatedly throughout the sequence. PMID- 12573244 TI - Familial hypofibrinogenaemia associated with heterozygous substitution of a conserved arginine residue; Bbeta255 Arg-->His (Fibrinogen Merivale). AB - Sequencing of all three fibrinogen genes from an individual with hypofibrinogenaemia led to the identification of two new point mutations in the Bbeta gene. Family studies showed the mutations Bbeta255 Arg-->His (Fibrinogen Merivale) and Bbeta148 Lys-->Asn (Fibrinogen Merivale II) were on different alleles and that only the Bbeta255 Arg-->His mutation segregated with hypofibrinogenaemia. Three simple heterozygotes for this mutation had mean fibrinogen concentrations of 1.4 mg/ml, while heterozygotes for the Bbeta148 Lys- >Asn mutation had normal fibrinogen concentrations. ESI MS analysis of endoproteinase Asp-N digests of Bbeta chains showed that the Bbeta255 Arg-->His substitution was not expressed in plasma, confirming it as the cause of the hypofibrinogenaemia. The Bbeta148 Lys-->Asn chains, on the other hand, were equally expressed with wild-type Bbeta chains in simple heterozygotes. Genotype analysis failed to detect either substitution in 182 healthy controls. Arg(255) is located in the first strand of the five-stranded sheet that forms the main feature of the betaD domain and appears to form an essential H bond with Gly(414). Both the Arg and Gly are absolutely conserved, not only in all known Bbeta chains, but also in all homologous alphaE and gamma chains and in all fibrinogen-related proteins. Protein instability from loss of this contact could easily explain the association of this mutation with hypofibrinogenaemia. PMID- 12573246 TI - Structural changes enhance the activity of Chainia xylanase in low urea concentrations. AB - Low concentrations of urea (1.2 M) stimulated the activity of endo-xylanase from Chainia by 30%. Subtle structural changes in the monomeric protein were reflected in the secondary and tertiary structure of the enzyme as monitored by fluorescence and circular dichroism. Changes in lambda(max) of emission, the fluorescence intensity and the Stern-Volmer quenching constants for acrylamide, measured in the presence of urea, indicated changes in the microenvironment of the Trp residues, suggesting alterations in tertiary structure. The ellipticity changes at 220 nm and Selcon analysis reflected changes in the content of beta sheet while both the near- and far-UV CD spectra indicated alterations in the secondary and tertiary structure of the protein in presence of urea. The dissociation constant values (K(d)) show very little change in the affinity of the enzyme for the substrate while the k(cat) values suggest enhanced turnover of the substrate in presence of urea. We suggest that low urea concentrations perturb the conformational state of xylanase leading to an open and a more flexible structure, resulting in enhanced catalytic rates. PMID- 12573247 TI - A Leu-Lys-rich antimicrobial peptide: activity and mechanism. AB - To develop novel antibiotic peptides useful as therapeutic drugs, the analogues were designed to increase not only net positive charge by Lys substitution but also hydrophobic helix region by Leu substitution from cecropin A (1-8)-magainin 2 (1-12) hybrid peptide (CA-MA). In particular, CA-MA analogue P5 (P5), designed by flexible region (GIG-->P) substitution, Lys (positions 4, 8, 14, 15) and Leu (positions 5, 6, 12, 13, 16, 17, 20) substitutions, showed an enhanced antimicrobial and antitumor activity without hemolysis. Confocal microscopy showed that P5 was located in the plasma membrane. The antibacterial effects of analogues were further confirmed by using 1,6-diphenyl-1,3,5-hexatriene as a plasma membrane probe. Flow cytometric analysis revealed that P5 acted in an energy-independent manner. This interaction is also independent of the ionic environment. Furthermore, P5 causes significant morphological alterations of the bacterial surfaces as shown by scanning electron microscopy and showed strong membrane disrupting activity when examined using liposomes (phosphatidyl choline/cholesterol; 10:1, w/w). Its potent antibiotic activity suggests that P5 is an excellent candidate as a lead compound for the development of novel antiinfective agents. PMID- 12573248 TI - Formation of functional heterodimers by isozymes 1 and 2 of pyruvate dehydrogenase kinase. AB - Pyruvate dehydrogenase kinase (PDK) is a mitochondrial enzyme responsible for regulation of the pyruvate dehydrogenase complex and, consequently, aerobic oxidation of carbohydrate fuels in general. In mammals, there are four genetically and biochemically distinct forms of PDK that are expressed in a tissue-specific manner (PDK1, PDK2, PDK3, and PDK4). These protein kinases have been shown to function as dimers, but the possibility of heterodimerization between various isozyme subunits has not yet been investigated. Here, we demonstrate that two members of the PDK family, PDK1 and PDK2, form heterodimeric species when coexpressed in the same Escherichia coli cell. The heterodimeric kinase produced in vivo was purified to near homogeneity by affinity chromatography. The purified kinase was stable and was not subjected to reassortment of the subunits. The heterodimeric kinase was catalytically active and was clearly distinct from homodimeric PDK1 or PDK2 with respect to kinetic parameters, site specificity and regulation. These data strongly suggest that heterodimerization between PDK1 and PDK2 adds another level of diversity to this protein family in addition to that which arises from gene multiplicity. PMID- 12573249 TI - Molecular characterization of a rabbit long-chain fatty acyl CoA synthetase that is highly expressed in the vascular endothelium. AB - The formation of coenzyme A thioesters from long-chain fatty acids represents a metabolic branch point. We have isolated, cloned and sequenced a long-chain fatty acyl CoA synthetase (LCFACoAS) that is localized to the endothelium of rabbit heart and aorta. Immunofluoresence and in situ hybridization studies show intense staining of the intimal layer of the aorta and coronary vessels. The microvessels, including the capillaries, of the coronary circulation also show intense immunofluoresence. The enzyme shares only about 30% to 70% homology with the primary amino acid sequence of the other known LCFACoAS. There is a region of 44 amino acids at the carboxy terminus, which is unique to the vascular enzyme. This domain contains the most hydrophobic region of the molecule, indicating that it may function as a membrane anchoring site. These results suggest that this LCFACoAS represents a novel isoform, whose functional significance remains to be determined. PMID- 12573250 TI - Tau could protect DNA double helix structure. AB - The hyperchromic effect has been used to detect the effect of tau on the transition of double-stranded DNA to single-stranded DNA. It was shown that tau increased the melting temperature of calf thymus DNA from 67 to 81 degrees C and that of plasmid from 75 to 85 degrees C. Kinetically, rates of increase in absorbance at 260 nm of DNA incubated with tau were markedly slower than those of DNA and DNA/bovine serum albumin used as controls during thermal denaturation. In contrast, rates of decrease in the DNA absorbance with tau were faster than those of controls when samples were immediately transferred from thermal conditions to room temperature. It revealed that tau prevented DNA from thermal denaturation, and improved renaturation of DNA. Circular dichroic spectra results indicated that there were little detectable conformational changes in DNA double helix when tau was added. Furthermore, tau showed its ability to protect DNA from hydroxyl radical (.OH) attacking in vitro, implying that tau functions as a DNA-protecting molecule to the radical. PMID- 12573251 TI - Chloroplast fructose 1,6-bisphosphatase with changed redox modulation: comparison of the Galdieria enzyme with cysteine mutants from spinach. AB - Spinach fructose 1,6-bisphosphatase (FBPase, EC 3.1.3.11), a redox-modulated chloroplast enzyme and part of the Calvin cycle, and three different Cys mutants were expressed in E. coli. The properties of the purified proteins were compared to those of native and recombinant chloroplast FBPase from the red alga Galdieria sulphuraria. In spinach chloroplast FBPase, Cys(155) and Cys(174) are engaged in the formation of the disulfide bridge. The corresponding mutants are active when expressed in E. coli, while C179S is inactive and can be reductively activated as can the wild-type enzyme. The active C174S mutant, however, could be inactivated by oxidation, and reactivated, but only by reduction, not alternatively with high pH and high Mg(2+) as is the case for the wild-type enzyme. In the sequence of Galdieria FBPase, the Cys that corresponds to Cys(179) in the spinach enzyme is lacking. However, the Galdieria FBPase, in contrast to the spinach Cys(179) mutant, does not show any indication for a comparable redox modulation of its activity. Instead, oxidation only leads to partial inactivation without any qualitative changes in enzyme properties. Upon reduction, the lost activity can be recovered. PMID- 12573253 TI - Optimized overproduction, purification, characterization and high-pressure sensitivity of the prion protein in the native (PrP(C)-like) or amyloid (PrP(Sc) like) conformation. AB - Overproduction and purification of the prion protein is a major concern for biological or biophysical analysis as are the structural specificities of this protein in relation to infectivity. We have developed a method for the effective cloning, overexpression in Escherichia coli and purification to homogeneity of Syrian golden hamster prion protein (SHaPrP(90-231)). A high level of overexpression, resulting in the formation of inclusion bodies, was obtained under the control of the T7-inducible promoter of the pET15b plasmid. The protein required denaturation, reduction and refolding steps to become soluble and attain its native conformation. Purification was carried out by differential centrifugation, gel filtration and reverse phase chromatography. An improved cysteine oxidation protocol using oxidized glutathione under denaturing conditions, resulted in the recovery of a higher yield of chromatographically pure protein. About 10 mg of PrP protein per liter of bacterial culture was obtained. The recombinant protein was identified by monoclonal antibodies and its integrity was confirmed by electrospray mass spectrometry (ES/MS), whereas correct folding was assessed by circular dichroism (CD) spectroscopy. This protein had the structural characteristics of PrP(C) and could be converted to an amyloid structure sharing biophysical and biochemical properties of the pathologic form (PrP(Sc)). The sensitivity of these two forms to high pressure was investigated. We demonstrate the potential of using pressure as a thermodynamic parameter to rescue trapped aggregated prion conformations into a soluble state, and to explore new conformational coordinates of the prion protein conformational landscape. PMID- 12573252 TI - Macrophage-induced rat mesangial cell expression of the 24p3-like protein alpha-2 microglobulin-related protein. AB - During screening of a murine macrophage cDNA repertoire for factors potentially able to modulate glomerular cell responses to injury, we identified a gene coding for the murine protein 24p3 lipocalin. Immunostaining of normal rat kidney sections showed positive 24p3-like staining in distal tubules/collecting ducts and small muscular arteries. Although most glomeruli were negative, some did exhibit small numbers of positively stained cells. Cultured rat glomeruli and glomerular mesangial cells secreted the 24p3-like protein in response to macrophage-conditioned medium (MPCM) and the cytokine IL-1beta. MPCM derived from TGFbeta-pretreated macrophages enhanced mesangial cell 24p3 secretion. In contrast, addition of anti-IL-1beta neutralising antibody to MPCM or IL-1beta resulted in suppression of 24p3 secretion. Co-culture of mesangial cells with varying numbers of non-LPS-treated macrophages resulted in dose-dependent secretion of 24p3 into culture supernatants. Archival sections from polyvinyl alcohol-treated and cholesterol-fed rats showed positive glomerular staining for 24p3 in and around glomerular foam cells. Nucleotide sequencing of rat mesangial cell-derived 24p3 cDNA revealed it to be identical to rat alpha-2-microglobulin related protein (alpha2microGRP), the rat homologue of murine 24p3. These data provide the first description of rat alpha2microGRP in the context of mesangial cell pathophysiology. PMID- 12573254 TI - In vitro lipolysis by human pancreatic lipase is specifically abolished by its inactive forms. AB - In human adults, the enzymatic hydrolysis of dietary fat along the digestive tract is sequentially catalyzed by two main enzymes, human gastric lipase (HGL) and human pancreatic lipase (HPL). Both a chemically inhibited form of HPL as well as an inactive HPL mutant with a glycine residue substituted for its catalytic serine were found to be strong inactivators of HPL activity. In the presence of bile salts, this inhibition was clearly due to competition for colipase. We established that the chemically inhibited HPL, probably in its open conformation, had a much greater affinity for colipase than the closed native form of HPL. These inhibitory effects are quite substantial, because a 0.2-M excess of the chemically inhibited HPL form relative to HPL reduced the catalytic lipolytic activity by 50% in the presence of an equimolar amount of colipase. PMID- 12573255 TI - Genomic basis of mucopolysaccharidosis type IIID (MIM 252940) revealed by sequencing of GNS encoding N-acetylglucosamine-6-sulfatase. AB - Mucopolysaccharidosis type IIID (MPS IIID; Sanfilippo syndrome type D; MIM 252940) is caused by deficiency of the activity of N-acetylglucosamine-6 sulfatase (GNS), which is normally required for degradation of heparan sulfate. The clinical features of MPS IIID include progressive neurodegeneration, with relatively mild somatic symptoms. Biochemical features include accumulation of heparan sulfate and N-acetylglucosamine-6-sulfate in the brain and viscera. To date, diagnosis required a specific lysosomal enzyme assay for GNS activity. From genomic DNA of a subject with MPS IIID, we amplified and sequenced the promoter and 14 exons of GNS. We found a homozygous nonsense mutation in exon 9 (1063C --> T), which predicted premature termination of translation (R355X). We also identified two common synonymous coding single-nucleotide polymorphisms and genotyped these in samples from four ethnic groups. This first report of a mutation in GNS resulting in MPS IIID indicates the potential utility of molecular diagnosis for this rare condition. PMID- 12573256 TI - Curly bare (cub), a new mouse mutation on chromosome 11 causing skin and hair abnormalities, and a modifier gene (mcub) on chromosome 5. AB - In the outcrossing of a new recessive mouse mutation causing hair loss, a new wavy-coated phenotype appeared. The two distinct phenotypes were shown to be alternative manifestations of the same gene mutation and attributable to a single modifier locus. The new mutation, curly bare (cub), was mapped to distal Chr 11 and the modifier (mcub) was mapped to Chr 5. When homozygous for the recessive mcub allele, cub/cub mice appear hairless. A single copy of the dominant Mcub allele confers a full, curly coat in cub/cub mice. Reciprocal transfer of full thickness skin grafts between mutant and control animals showed that the skin phenotype was tissue autonomous. The hairless cub/cub mcub/mcub mice show normal contact sensitivity responses to oxazolone. The similarity of the wavy coat phenotype to those of Tgfa and Egfr mutations and the map positions of cub and mcub suggest candidate genes that interact in the EGF receptor signal transduction pathway. PMID- 12573257 TI - NECC1, a candidate choriocarcinoma suppressor gene that encodes a homeodomain consensus motif. AB - We isolated a candidate choriocarcinoma suppressor gene from a PCR-based subtracted fragmentary cDNA library between normal placental villi and the choriocarcinoma cell line CC1. This gene comprises an open reading frame of 219 nt encoding 73 amino acids and contains a homeodomain as a consensus motif. This gene, designated NECC1 (not expressed in choriocarcinoma clone 1), is located on human chromosome 4q11-q12. NECC1 expression is ubiquitous in the brain, placenta, lung, smooth muscle, uterus, bladder, kidney, and spleen. Normal placental villi expressed NECC1, but all choriocarcinoma cell lines examined and most of the surgically removed choriocarcinoma tissue samples failed to express it. We transfected this gene into choriocarcinoma cell lines and observed remarkable alterations in cell morphology and suppression of in vivo tumorigenesis. Induction of CSH1 (chorionic somatomammotropin hormone 1) by NECC1 expression suggested differentiation of choriocarcinoma cells to syncytiotrophoblasts. Our results suggest that loss of NECC1 expression is involved in malignant conversion of placental trophoblasts. PMID- 12573258 TI - SLURP-2, a novel member of the human Ly-6 superfamily that is up-regulated in psoriasis vulgaris. AB - By microarray assay we identified ESTs (expressed sequence tags) whose expression was predominantly increased in the affected skin of patients with psoriasis vulgaris. Among them, a full-length cDNA sequence corresponding to one of those ESTs (AI829641) was isolated by screening of cultured human keratinocyte cDNA libraries. This cDNA encodes a novel member of the Ly-6/uPAR superfamily, designated SLURP-2 (secreted Ly-6/uPAR related protein 2). SLURP-2 has an open reading frame of 97 amino acids containing 10 conserved cysteine residues. SLURP 2 has a single functional copy within the LY6 superfamily gene cluster at chromosome 8q24.3. RT-PCR (reverse transcriptase-polymerase chain reaction) expression analysis revealed that SLURP-2 was expressed in multiple tissues, mainly in the epithelial cells including the skin and keratinocytes, but not in spleen or bone marrow. Comparison of the expression of this gene among the psoriatic lesional and nonlesional skin of patients and the normal skin of healthy individuals detected by quantitative real-time RT-PCR analysis disclosed that SLURP-2 was up-regulated threefold in psoriatic lesional skin. These findings suggest that SLURP-2 may be involved in the pathophysiology of psoriasis through its role in keratinocyte hyperproliferation and/or T cell differentiation/activation. PMID- 12573259 TI - Mouse Wnt9b transforming activity, tissue-specific expression, and evolution. AB - The members of the Wnt family of secreted factors have oncogenic potential and important roles as developmental regulators. We report an analysis of mouse Wnt9b (also called Wnt15 and Wnt14b), including its cDNA sequence, chromosomal mapping, epithelial cell transforming activity, adult and embryonic tissue expression patterns, and evolution. We also deduced the full-length amino acid sequence of its close relative, Wnt9a (also called Wnt14), from unannotated genomic DNA sequences in GenBank. Full-length comparisons among Wnt amino acid sequences provide evidence that Wnt9b and Wnt9a are close paralogs of each other and are orthologs of Wnt9 genes from shark and hagfish. Mapping Wnt9b to The Jackson Laboratory BSS interspecific backcross panel places it at 63.0 cM on chromosome 11. Sequence comparisons of two pairs of linked Wnt genes (the Wnt9a-Wnt3a pair and the Wnt9b-Wnt3 pair) suggest that they arose from the relatively recent duplication of a single ancestral Wnt gene pair, confirming the close paralogous relationship of Wnt9a and Wnt9b. Wnt9b expression is primarily restricted to the kidney in the adult mouse, with lower levels detected in the preputial gland, liver, and mammary gland. Testing of staged whole mouse embryos from 9.5 to 17.5 days of gestation showed expression at all stages with a peak at day 10.5. In situ hybridization analysis showed expression in most but not all tissues of the 16.5-day embryo. No significant elevation of Wnt9b expression was detected in 29 mouse mammary tumor virus-induced tumors. Overexpression of Wnt9b in C57MG mammary epithelial cells caused small transformed foci in cell monolayers and a moderate morphological transformation in pooled colonies compared with Wnt1. PMID- 12573260 TI - Characterization of the genomic structure and expression of the mouse Apex2 gene. AB - We isolated a mouse cDNA encoding APEX2 protein and demonstrated that APEX2 binds to PCNA. The level of Apex2 mRNA was high in the thymus, bone marrow, spleen, and kidney in adult mice. Apex2 consists of six exons and is flanked on the 3' end by Alas2 on X chromosome 63.0. Furthermore, Apex2 is flanked on the 5' end by a novel gene with a 106-bp intergenic sequence. We disrupted Apex2 in embryonic stem cells derived from a male mouse, and a 55-kDa APEX2 protein was detected in the nuclei of Apex2(+) but not Apex2-disrupted cells. Immunoelectron microscopy revealed that APEX2 is also localized in the mitochondria of Apex2(+) cells. In serum-stimulated BALB/c 3T3 cells, the level of Apex2 mRNA was transiently increased and the level of APEX2 reached a maximum in the late S phase, thus indicating that APEX2 may participate in postreplicative base excision repair. PMID- 12573261 TI - Antisense transcripts at the EMX2 locus in human and mouse. AB - The homeodomain transcription factor EMX2 is critical for central nervous system and urogenital development. In addition, EMX2 maps to a region of allelic deletion corresponding to a putative endometrial tumor suppressor at 10q26. We now report another polyadenylated transcript that is transcribed on the strand opposite to EMX2 and overlaps with the EMX2 transcript. This transcript was designated EMX2OS (OS, opposite strand), and an orthologous transcript present at the murine Emx2 locus was designated Emx2os. Alternative splicing to generate transcripts with varying 5' sequences was detected in the human but not the mouse. Neither ortholog contains a significant open reading frame, nor is primary sequence conserved between the two species. The sense and antisense transcripts display coordinate expression in that EMX2 and EMX2OS are abundant in normal postmenopausal endometrium, reduced in premenopausal endometrium, and absent or reduced in a majority of primary endometrial tumors. EMX2, EMX2OS, Emx2, and Emx2os are abundant in the uterine endometrium, with sense and antisense transcripts exhibiting identical expression patterns. Conservation of functional human and murine EMX2 antisense genes, of overlap between the sense and the antisense transcripts, and of identical cellular expression patterns suggests a biological function for EMX2OS, presumably to regulate EMX2. PMID- 12573262 TI - Evolutionary constraints on the Disrupted in Schizophrenia locus. AB - The Disrupted in Schizophrenia (DISC) locus on human chromosome 1q42 has been strongly implicated by genetic studies as a susceptibility locus for major mental illnesses. In humans the locus is transcriptionally complex, with multiple alternate splicing events, antisense transcription, and intergenic splicing all evident. We have compared the genomic sequence and transcription maps of this locus between human, mouse, pufferfish (Fugu rubripes), and, in part, zebrafish (Danio rerio). The order and orientation of EGLN1, TSNAX, and DISC1 genes are conserved between mammals and F. rubripes. Intergenic splicing and short intergenic transcripts are not found to be conserved features. DISC2, a putative noncoding transcript partially antisense to DISC1, is not conserved in mouse or F. rubripes. Alternate splice forms of the protein-coding DISC1 gene are conserved even though the genomic structure is not. The amino acid sequence of DISC1 is diverging rapidly, although a putative nuclear localization signal and discrete blocks of coiled coil are specifically conserved features. PMID- 12573263 TI - Evolution of the cystatin B gene: implications for the origin of its variable dodecamer tandem repeat in humans. AB - The human cystatin B gene contains a variable number of 12-bp tandem repeats in its promoter region, of which the common alleles contain two or three copies and unusual expansion causes progressive myoclonus epilepsy of the Unverricht Lundborg type. We undertook a comprehensive analysis of the genomic sequence to address the evolutionary events of this variable repeat. By examination of a contiguous genome sequence spanning 5.0 kb and linkage analysis of detected polymorphic changes, we identified six major intragenic haplotypes in unrelated Japanese subjects. The number of normal repeats was closely correlated with these alleles, indicating that changes in the array should be comparatively rare events during human evolution. To examine the origin of the repeat array further, we also analyzed five primate genomes. Repetitive polymorphism was unlikely in hominoids, and the array originated with the dodecamer itself in the course of primate evolution. The variability conceivably developed after the separation to humans. PMID- 12573265 TI - OPCAB surgery: a voyage of discovery back to the future. Off-pump coronary artery bypass. PMID- 12573264 TI - Single-nucleotide polymorphisms in the Toll-like receptor 9 gene (TLR9): frequencies, pairwise linkage disequilibrium, and haplotypes in three U.S. ethnic groups and exploratory case-control disease association studies. AB - TLR9 is a mammalian Toll-like receptor homologue that appears to function as an innate immune pattern recognition protein for motifs that are far more common in bacterial than in mammalian DNA. The gene was sequenced in 71 subjects from three self-identified U.S. ethnic groups to identify single-nucleotide polymorphisms (SNPs). A total of 20 SNPs were found of which only 20% were in the public dbSNP database. Four SNPs were relatively common in all three ethnic samples. Using these four SNPs, seven distinct haplotypes were statistically inferred, of which four accounted for 75% or more chromosomes. These four haplotypes could be distinguished from each other by the alleles of two SNPs (-1237 and 2848). Five exploratory nested case-control disease-association studies (asthma, DVT, MI, and COPD in European Americans and asthma in African Americans) were performed by genotyping DNA collected from four ongoing cohort studies. There was evidence suggesting increased risk for asthma with a C allele at -1237 (odds ratio 1.85, 95%CI 1.05 to 3.25) among European Americans (genotypes available from 67 cases and 152 controls). No other significant disease associations were detected. Replication of this finding in other, larger samples is needed. This study suggests that there is substantial diversity in human TLR9, possibly associated with asthma in Europeans but not African Americans. No association was detected with three other diseases potentially related to innate immunity. PMID- 12573266 TI - Angiotensin receptor blockers and clinical trials in heart failure. PMID- 12573267 TI - Singapore and coronary heart disease: a population laboratory to explore ethnic variations in the epidemiologic transition. PMID- 12573268 TI - Closure of atrial septal defects: is the debate over? PMID- 12573269 TI - Aortic stenosis. PMID- 12573270 TI - Cardiac markers in acute coronary syndromes-refining our knowledge. PMID- 12573271 TI - The challenge of in-stent restenosis: insights from intravascular ultrasound. PMID- 12573272 TI - Ethnic differences in acute myocardial infarction in Singapore. AB - AIMS: We compare the myocardial infarction (MI) event and mortality rates among Chinese, Malay and Indian residents of Singapore. METHODS: Residents, aged 20 to 64 years, with an MI event were identified from hospital discharge listings, postmortem reports, and the Registry of Births and Deaths. All pathology laboratories flagged patients with elevated creatine phosphokinase (CPK) levels. Modified MONICA (multinational monitoring of trends and determinants in cardiovascular disease) criteria were used for determining MI events. RESULTS: From 1991 to 1999, 12 481 MI events were identified. Chinese patients were older and less likely to have typical symptoms or previous MI. Malays had the highest peak CPK level. Among all three ethnic groups, MI event and age-adjusted case fatality rates declined. Compared with Chinese, MI event rates were >2-fold and >3-fold higher, and age-standardized coronary mortality rates were 2.4 and 3.0 higher times for Malays and Indians, respectively. Malays have the highest 3.1 year case-fatality, with an adjusted hazard ratio of 1.26 (95% confidence interval, 1.14 to 1.38) compared with Chinese. CONCLUSION: We found strong ethnic differences in MI event, case-fatality and coronary mortality rates among the three ethnic groups in Singapore. While Indians have the greatest MI event rates, Malays have the highest case-fatality. PMID- 12573273 TI - Impact of pre-hospital care in patients with acute myocardial infarction compared with those first managed in-hospital. AB - AIMS: To compare prospectively the impact of pre-hospital care by a physician staffed mobile coronary care unit with patients managed initially in-hospital, all with acute myocardial infarction. METHODS AND RESULTS: This was a single centre registry of consecutive patients (n=750) admitted with acute myocardial infarction to the coronary care unit and cardiology wards of the Royal Victoria Hospital, Belfast between 1998 and 2001. For the 750 patients, in-hospital mortality was 11% and was significantly lower for those managed pre-hospital (8% vs 13%, P=0.04): patients who received fibrinolytic therapy (n=474), the in hospital mortality was significantly lower in the pre-hospital group (7% vs 13%, P=0.02). Those managed pre-hospital had significant reduction in the median delay times (25th, 75th percentiles) from onset of symptoms to call for help 1.0 (0.5, 2.2) vs 2.0 (0.9, 6.0) h, P<0.001, from call for help to receiving fibrinolytic therapy 1.0 (0.8, 1.5) vs 1.8 (1.2, 2.5) h, P<0.001 resulting in a shorter pain to-needle time for fibrinolytic therapy 2.3 (1.5, 3.8) vs 4.0 (2.6, 7.2) h, P<0.001. For all patients, older age, haemodynamic indicators on admission (hypotension, higher heart rate, heart failure) and managed by the in-hospital route were significant independent variables for an adverse in-hospital mortality. Although for patients aged >or=75 years no statistical significant reduction in mortality occurred for those managed pre-hospital (P=0.051), nevertheless patients in this age group first treated pre-hospital who received fibrinolytic therapy had a significantly lower mortality than those first treated in-hospital (21% vs 43%, P=0.02). CONCLUSIONS: Consecutive patients with acute myocardial infarction seen and managed initially out-of-hospital by a physician staffed mobile coronary care unit had significantly lower in-hospital mortality. PMID- 12573274 TI - Predictors of 90-day outcome in patients stabilized after acute coronary syndromes. AB - AIMS: We investigated predictors of 90-day risk among patients surviving the early period after an acute coronary syndrome (ACS). METHODS AND RESULTS: The study population included 15 904 stabilized ST-segment elevation or non-ST segment elevation ACS patients randomized in SYMPHONY and 2nd SYMPHONY. We developed risk models for death, death or myocardial infarction (MI), and death, MI, or severe recurrent ischaemia (SRI) using Cox proportional-hazards techniques. Demographic, history, and pre-randomization clinical and medication variables were tested. Validation techniques included development of individual trial models, backward elimination and bootstrapping. Of 118 variables, 17 independently predicted mortality. The strongest associations included greater age (chi(2)=31.1), higher randomization heart rate (chi(2)=27.4), and heart failure (HF) variables (HF between qualifying event and randomization, chi(2)=21.8; history of HF, chi(2)=12.2). Higher creatinine clearance (chi(2)=17.7) and percutaneous coronary intervention between qualifying event and randomization (chi(2)=11.1) most strongly predicted lower risk. Similar characteristics entered the double and triple composite models, but HF variables and age less strongly predicted these end-points. CONCLUSIONS: In patients stabilized after ACS, those at highest risk over the next 90 days can be identified. Typical clinical markers are better at identifying risk of death than non-fatal MI or SRI. Novel risk markers are needed for these outcomes. PMID- 12573275 TI - Time-based risk assessment after myocardial infarction. Implications for timing of discharge and applications to medical decision-making. AB - AIMS: We evaluated timing of adverse cardiac events after thrombolysis to guide length of stay after ST-segment elevation myocardial infarction. METHODS AND RESULTS: Kaplan-Meier survival curves described timing of major postinfarction complications in 41021 fibrinolytic-treated patients in GUSTO-I. Using model fitting, these data were best explained by a mixed-exponential survival model: an acute curve describing most adverse events and a chronic curve describing a lower background rate. We replicated this strategy in 15059 fibrinolytic-treated patients in GUSTO-III. From the relation between time and events described by the model's acute curve in GUSTO-III, we proposed times for hospital discharge. The acute curve explained 97% of deaths and 68%-96% of various event composites. Of complications within 10 days, 90% of deaths and 70% of acute curve death, stroke, shock, heart failure, or reinfarction occurred by 24 h. By 2.7 days, 95% of deaths, stroke, shock, heart failure, or reinfarction occurred. Most major ventricular arrhythmias occurred within 24 h, after which the hazard curve was flat. CONCLUSIONS: Mixed-exponential survival modelling describes timing of post infarction complications and supports discharge 4 days after uncomplicated infarction. Such time-based risk assessment could guide decision-making in other settings in which randomized studies are impractical. PMID- 12573276 TI - Excellent survival and low incidence of arrhythmias, stroke and heart failure long-term after surgical ASD closure at young age. A prospective follow-up study of 21-33 years. AB - AIMS: Although studies have suggested good long-term results, arrhythmias, pulmonary hypertension and left ventricular dysfunction are mentioned as sequelae long-term after surgical atrial septal defect closure at young age. Most studies were performed only by questionnaire and in a retrospective manner. The long-term outcome is very important with regard to future employment and acceptance on insurance schemes. METHODS AND RESULTS: One hundred and thirty-five consecutive ASD-patients, operated on in childhood, were studied longitudinally with ECG, echocardiography, exercise testing and Holter-recording 15 (10-22) and 26 (21-33) years after surgery. During follow-up no cardiovascular mortality, stroke, heart failure and no pulmonary hypertension occurred. Symptomatic supraventricular tachyarrhythmias were present in 6% after 15 years, and an additional 2% occurred in the last decade; 5% needed pacemaker implantation. No relation was found between arrhythmias and type of ASD, baseline data, right ventricular dimensions, or age at operation. Left and right ventricular function and dimension remained unchanged. Slightly more patients had right atrial dilatation at last follow-up. Exercise capacity was comparable with the normal Dutch population. CONCLUSIONS: The long-term outcome after ASD closure at young age shows excellent survival and low morbidity. The incidence of supraventricular arrhythmias is lower than in natural history studies of ASD patients and also lower than after surgical correction at adult age. PMID- 12573277 TI - Lipoprotein(a), Chlamydia pneumoniae, leptin and tissue plasminogen activator as risk markers for valvular aortic stenosis. AB - AIMS: The aim of the present study was to identify risk markers for the development of valvular aortic stenosis (AS). Lipoprotein(a) (Lp(a)) and Chlamydia pneumoniae IgG antibody titres in plasma and in circulating immune complexes as well as leptin and tissue plasminogen activator (t-PA) in plasma were studied. METHODS AND RESULTS: One hundred and one patients (41 women and 60 men, mean age 71+/-8 years) with significant AS and 101 age- and sex-matched controls were included in this study. All patients underwent aortic valve replacement at the University Hospital in Umea, Sweden. The controls had no symptoms of cardiovascular disease and they were examined echocardiographically. An Lp(a) level >or=480 mg x l(-1), a C. pneumoniae-specific IgG titre >or=1/128, a high leptin level and a high t-PA mass concentration in plasma were identified as risk markers for AS. A strong synergism between Lp(a) and C. pneumoniae IgG antibodies in circulating immune complexes was found. CONCLUSION: Our data indicate that a chronic C. pneumoniae infection and a high plasma Lp(a) level might influence and aggravate aortic heart valve sclerosis via the formation of circulating immune complexes. The present study also strongly suggests an association between high plasma leptin, t-PA mass concentration and AS. PMID- 12573278 TI - Alternatively spliced FGFR-1 isoform signaling differentially modulates endothelial cell responses to peroxynitrite. AB - Mounting experimental evidence has suggested that the trophic environment of cells in culture is an important determinant of their vulnerability to the cytotoxic effects of reactive oxidants such as peroxynitrite (ONOO(-)). However, acidic fibroblast growth factor (FGF-1)-induced signaling renders some cells more sensitive and others resistant to the cytotoxic effects of ONOO(-). To determine whether alternatively spliced fibroblast growth factor receptor (FGFR-1) isoforms are responsible for this differential response, we have stably transfected FGFR negative rat brain-derived resistant vessel endothelial cells (RVEC) with human cDNA sequences encoding either FGFR-1 alpha or FGFR-1 beta. FGF-1 treatment of RVEC(R-1 alpha) transfectants enhanced ONOO(-)-mediated cell death in a manner dependent upon FGFR-1 tyrosine kinase, MEK/Erk 1/2 kinase, and p38 MAP kinase activities and independent of Src-family kinase (SFK) activity. FGF-1 treatment of RVEC(R-1 beta) transfectants inhibited the cytotoxic effects of ONOO(-) in a manner dependent upon FGFR-1 tyrosine kinase, MEK/Erk 1/2 kinase, and SFK activities and independent of p38 MAP kinase activity. FGF-1-induced preactivation of both FGFR-1 tyrosine and Erk 1/2 kinases was detected in both RVEC(R-1 alpha) and RVEC(R-1 beta) transfectants. FGF-1-induced preactivation of p38 MAPK was restricted to RVEC(R-1 alpha) transfectants, whereas, ligand-induced preactivation of SFK was limited to RVEC(R-1 beta) transfectants. Collectively, these results both reemphasize the role of extracellular trophic factors and their receptor-mediated signaling pathways during cellular responses to oxidant stress and provide a first indication that the alternatively spliced FGFR-1 isoforms induce differential signal transduction pathways. PMID- 12573279 TI - Cytochrome c-induced cytosolic nicotinamide adenine dinucleotide oxidation, mitochondrial permeability transition, and apoptosis. AB - A catalytic amount of cytochrome c (cyto-c) added to the incubation medium of isolated mitochondria promotes the transfer of reducing equivalents from extramitochondrial nicotinamide adenine dinucleotide in its reduced state (NADH) to molecular oxygen inside the mitochondria, a process coupled to the generation of a membrane potential. This mimics in many aspects the early stages of those apoptotic pathways characterized by the persistence of mitochondrial membrane potential but with cyto-c already exported into the cytosol. In cyclosporin sensitive and calcium-induced mitochondrial permeability transition (MPT) a release of cyto-c can also be observed. However, in MPT uncoupled respiration associated with mitochondrial swelling and preceded by the complete dissipation of the membrane potential which cannot be restored with ATP addition or any other source of energy is immediately activated. The results obtained and discussed with regard to intactness of mitochondrial preparations indicate that MPT could be an apoptotic event downstream but not upstream of cyto-c release linked to the energy-requiring processes. In the early stages of apoptosis cytosolic cyto-c participates in the activation of caspases and at the same time can promote the oxidation of cytosolic NADH, making more energy available for the correct execution of the cell death program. This hypothesis is not in contrast with available data in the literature showing that cyto-c is present in the cytosol of both control and apoptosis-induced cultured cell lines. PMID- 12573280 TI - Intracellular mechanisms of hypoxia-induced calcium increase in rat sensory neurons. AB - Elevation of cytosolic level of Ca(2+) was measured by spatial screening of freshly isolated dorsal root ganglion neurons loaded with Fura-2AM after subjecting them to a moderate hypoxic solution (pO(2)=10-40 mmHg). Short exposure of neurons to hypoxia resulted in a reversible elevation of intracellular Ca(2+) to about 120% in the cell center and to 80% in the cell periphery. Such elevation could be almost completely eliminated by removal of Ca(2+) or Na(+) from external medium or application of nifedipine, an L-type calcium channel blocker. Remarkable antihypoxic efficiency (58%) was achieved by preapplication of mitochondrial protonophore CCCP. A conclusion is made that in sensory neurons the hypoxia-induced elevation of cytosolic Ca(2+) is induced by combined changes of function in three cell substructures: voltage-operated L-type Ca(2+) and Na(+) channels and Ca(2+) accumulation by mitochondria. Mitochondria are important for spatial difference in the hypoxia-induced Ca(2+) elevation due to their specific location in these neurons. PMID- 12573281 TI - Isolation, purification, and physicochemical characterization of a D-galactose binding lectin from seeds of Erythrina speciosa. AB - A lectin was isolated from the saline extract of Erythrina speciosa seeds by affinity chromatography on lactose-Sepharose. The lectin content was about 265 mg/100g dry flour. E. speciosa seed lectin (EspecL) agglutinated all human RBC types, showing no human blood group specificity; however a slight preference toward the O blood group was evident. The lectin also agglutinated rabbit, sheep, and mouse blood cells and showed no effect on horse erythrocytes. Lactose was the most potent inhibitor of EspecL hemagglutinating activity (minimal inhibitory concentration (MIC)=0.25 mM) followed by N-acetyllactosamine, MIC=0.5mM, and then p-nitrophenyl alpha-galactopyranoside, MIC=2 mM. The lectin was a glycoprotein with a neutral carbohydrate content of 5.5% and had two pI values of 5.8 and 6.1 and E(1%)(1 cm) of 14.5. The native molecular mass of the lectin detected by hydrodynamic light scattering was 58 kDa and when examined by mass spectroscopy and SDS-PAGE it was found to be composed of two identical subunits of molecular mass of 27.6 kDa. The amino acid composition of the lectin revealed that it was rich in acidic and hydroxyl amino acids, contained a lesser amount of methionine, and totally lacked cysteine. The N-terminal of the lectin shared major similarities with other reported Erythrina lectins. The lectin was a metaloprotein that needed both Ca(2+) and Mn(2+) ions for its activity. Removal of these metals by EDTA rendered the lectin inactive whereas their addition restored the activity. EspecL was acidic pH sensitive and totally lost its activity when incubated with all pH values between pH 3 and pH 6. Above pH 6 and to pH 9.6 there was no effect on the lectin activity. At 65 degrees C for more than 90 min the lectin was fairly stable; however, when heated at 70 degrees C for 10 min it lost more than 80% of its original activity and was totally inactivated at 80 degrees C for less than 10 min. Fluorescence studies of EspecL indicated that tryptophan residues were present in a highly hydrophobic environment, and binding of lactose to EspecL neither quenched tryptophan fluorescence nor altered lambda(max) position. Treating purified EspecL with NBS an affinity-modifying reagent specific for tryptophan totally inactivated the lectin with total modification of three tryptophan residues. Of these residues only the third modified residue seemed to play a crucial role in the lectin activity. Addition of lactose to the assay medium did not provide protection against NBS modification which indicated that tryptophan might not be directly involved in the binding of haptenic sugar D-galactose. Modification of tyrosine with N-acetylimidazole led to a 50% drop in EspecL activity with concomitant acetylation of six tyrosine residues. The secondary structure of EspecL as studied by circular dichroism was found to be a typical beta-pleated-sheet structure which is comparable to the CD structure of Erythrina corallodendron lectin. Binding of lactose did not alter the EspecL secondary structure as revealed by CD examination. PMID- 12573282 TI - Characterization of the interaction of Rhodobacter capsulatus cytochrome c peroxidase with charge reversal mutants of cytochrome c(2). AB - Steady-state kinetics for the reaction of Rhodobacter capsulatus bacterial cytochrome c peroxidase (BCCP) with its substrate cytochrome c(2) were investigated. The Rb. capsulatus BCCP is dependent on calcium for activation as previously shown for the Pseudomonas aeruginosa BCCP and Paracoccus denitrificans enzymes. Furthermore, the activity shows a bell-shaped pH dependence with optimum at pH 7.0. Enzyme activity is greatest at low ionic strength and drops off steeply as ionic strength increases, resulting in an apparent interaction domain charge product of -13. All cytochromes c(2) show an asymmetric distribution of surface charge, with a concentration of 14 positive charges near the exposed heme edge of Rb. capsulatus c(2) which potentially may interact with approximately 6 negative charges, localized near the edge of the high-potential heme of the Rb. capsulatus BCCP. To test this proposal, we constructed charge reversal mutants of the 14 positively charged residues located on the front face of Rb. capsulatus cytochrome c(2) and examined their effect on steady-state kinetics with BCCP. Mutated residues in Rb. capsulatus cytochrome c(2) that showed the greatest effects on binding and enzyme activity are K12E, K14E, K54E, K84E, K93E, and K99E, which is consistent with the site of electron transfer being located at the heme edge. We conclude that a combination of long-range, nonspecific electrostatic interactions as well as localized salt bridges between, e.g., cytochrome c(2) K12, K14, K54, and K99 with BCCP D194, D241, and D6, account for the observed kinetics. PMID- 12573283 TI - The physico-chemical characterization of a boiling stable antifreeze protein from a perennial grass (Lolium perenne). AB - We have characterized a cold-induced, boiling stable antifreeze protein. This highly active ice recrystallization inhibition protein shows a much lower thermal hysteresis effect and displays binding behavior that is uncharacteristic of any AFP from fish or insects. Ice-binding studies show it binds to the (1 0 1 0) plane of ice and FTIR studies reveal that it has an unusual type of highly beta sheeted secondary structure. Ice-binding studies of both glycosylated and nonglycosylated expressed forms indicate that it adsorbs to ice through the protein backbone. These results are discussed in light of the currently proposed mechanisms of AFP action. PMID- 12573284 TI - Cationic liposome-mediated gene delivery: biophysical study and mechanism of internalization. AB - To identify factors affecting cationic liposome-mediated gene delivery efficiency, we studied the relationship between the biophysical characteristics of liposome/DNA complexes (lipoplexes) at different (+/-) charge ratios, their structures as monitored by atomic force microscopy (AFM), and their mechanism(s) of internalization into the cells. Significant changes were observed in the particle size and zeta potential of liposomes and their structures assessed by AFM upon addition of DNA, which depended on (+/-) charge ratios. AFM images showed that lipoplexes were formed from extensively fused and apparently homogeneous lipid particles encapsulating DNA. Lipoplexes were found to internalize the cells through the endocytosis pathway. Lipoplex-cell fusion was found to occur mainly at the plasma membrane level; however, this lipoplex-cell membrane fusion was found to be essential for the uptake of the large particles. A new perspective for the internalization of large lipoplex particles into cytoplasm is discussed. PMID- 12573285 TI - Identification and solution conformation of multiple epitopes recognized by a MUC2 mucin-specific monoclonal antibody. AB - We have identified the optimal epitope, 21TQTPT25, in the tandem repeat of mucin 2 (MUC2) glycoprotein by using glycoprotein-specific monoclonal antibody, MAb 994, and synthetic, overlapping and truncated oligopeptides corresponding to the sequence 13TPTPTPTGTQTPTT26. We found that peptides containing the 21TQTPT25 sequence were able to inhibit the 994 antibody binding and also peptides 21TQTPT25 and 17TPTGTQTPT25 were the most inhibitory compounds with the lowest IC50 value (IC50=4 and 3 microM, respectively) tested. Interestingly, 21TQTPT25 peptide adopts an unordered structure even in TFE, a solvent that promotes an ordered conformation, as detected by circular dichroism and Fourier-transform infrared spectroscopy. However, Thr at position 26 or amidation of Thr25 at the C terminus results in a much weaker (3 orders of magnitude) MAb interaction, which can be due to the presence of a turn conformation in peptides with a T26 or an amide C-terminus. We have also observed that MAb 994 recognized two other pentapeptides with the TX1TX2T motif, like 13TPTPT17 (IC50=180 microM) and 19TGTQP23 (IC50=65 microM), whose sequences are present in the native glycoprotein. These findings might suggest that in the MUC2 tandem repeat unit there are multiple antigenic sites available for recognition in underglycosylated tumor tissue and also explain the heteroclitic nature of MAb 994. PMID- 12573286 TI - Sequence-specific DNA damage induced by ultraviolet A-irradiated folic acid via its photolysis product. AB - DNA damage mediated by photosensitizers participates in solar carcinogenesis. Fluorescence measurement and high-performance liquid chromatography analysis demonstrated that photoirradiated folic acid, one of the photosensitizers in cells, generates pterine-6-carboxylic acid (PCA). Experiments using 32P-labeled DNA fragments obtained from a human gene showed that ultraviolet A-irradiated folic acid or PCA caused DNA cleavage specifically at consecutive G residues in double-stranded DNA after Escherichia coli formamidopyrimidine-DNA glycosylase or piperidine treatment. The amount of 8-oxo-7,8-dihydro-2(')-deoxyguanosine formed through this DNA photoreaction in double-stranded DNA exceeded that in single stranded DNA. Kinetic studies suggested that DNA damage is caused mainly by photoexcited PCA generated from folic acid rather than by folic acid itself. In conclusion, photoirradiated folic acid generates PCA, which induces DNA photooxidation specifically at consecutive G residues through electron transfer. Excess intake of folic acid supplements may increase a risk of skin cancer by solar ultraviolet light. PMID- 12573288 TI - Thermodynamics of the reduction of NADP with 2-propanol catalyzed by an NADP dependent alcohol dehydrogenase. AB - The apparent equilibrium constant of the biochemical reaction, 2 propanol+NADP(ox) = acetone+NADP(red), was determined at I = 0.25 M over a wide range of pH (5.63 to 8.02) and temperature (5 to 40 degrees C). The reaction was catalyzed by an NADP-dependent alcohol dehydrogenase. The results were used to calculate thermodynamic quantities for the chemical (ionic) reference reaction: 2 propanol+NADP(ox)(3-) = acetone+NADP(red)(4-)+H(+). The thermodynamic quantities for this reference reaction are as follows: equilibrium constant K = (5.98+/ 0.46) x 10(-10); standard molar Gibbs energy change Delta(r)G(0) = (52.65+/-0.19) kJmol(-1); standard molar enthalpy change Delta(r)H(0) = (38.9+/-0.6) kJmol(-1); and standard molar entropy change Delta(r)S(0) = -(46.1+/-2.2)J K(-1)mol(-1). All of these results pertain to 25 degrees C (298.15 K) and I = 0. The results also lead, in conjunction with tabulated thermodynamic quantities, to the standard electromotive force E(0) = -0.140 V for the reduction of NADP(ox)(3-) to NADP(red)(4-). PMID- 12573287 TI - Regulation of protein tyrosine phosphatase 1B in intact cells by S-nitrosothiols. AB - Protein tyrosine phosphatases (PTPases) contain an active site cysteine which when oxidized leads to loss of phosphatase activity and accumulation of phosphoproteins. For example, oxidants produced following EGF stimulation inhibit PTP1B and enhance EGF receptor phosphorylation. Because NO-derived species also modify reactive thiols, we postulated that NO would reversibly inhibit PTP1B. In our studies we exposed A431 or Jurkat cells to NO donors and measured PTP1B activity or used 3-maleimidylpropionylbiocytin (MPB) to measure thiol redox status. Nitrosothiols led to a rapid inhibition of PTP1B through a mechanism that was greatly enhanced by addition of cysteine to the medium. Analysis of thiol oxidation status using immunoprecipitated PTP1B showed modification consistent with loss of activity. Both enzyme inhibition and modification were reversible in intact cells or after addition of DTT to cell lysates. While DTT reversed oxidation, ascorbate did not, suggesting that formation of a mixed disulfide (possibly glutathionylation) rather than S-nitrosylation accounts for PTP1B inhibition. Importantly, PTP1B inhibition by nitrosothiols led to EGF receptor phosphorylation even in the absence of exogenously added EGF. These findings suggest an important role for NO in modulating signaling pathways since inhibition of PTPases could potentially enhance or prolong activity of phosphoproteins. PMID- 12573289 TI - Mechanism of melatonin-induced oscillations in the peroxidase-oxidase reaction. AB - Melatonin induces oscillations in the peroxidase-oxidase (PO) reaction catalyzed by horseradish peroxidase. We present here studies of the effect of pH, enzyme concentration, and concentration of melatonin on the oscillation frequency. We also present a mechanistic model to explain the experimentally observed changes in oscillation frequency. Using the data obtained here we are able to predict that oscillations will also occur in the PO reaction catalyzed by myeloperoxidase. Myeloperoxidase is an important protein in activated neutrophils and we provide evidence that the oscillations of NAD(P)H, superoxide and hydrogen peroxide in these cells may involve this enzyme. Thus, our experimental system can be considered a model system for the nonrespiratory oxygen metabolism in activated neutrophils and other similar cells participating in the defence against invading pathogens. PMID- 12573290 TI - Relationship of chloroquine-induced redistribution of a neutral aminopeptidase to hemoglobin accumulation in malaria parasites. AB - To study the relationship between neutral aminopeptidase activity and hemoglobin accumulation in malaria parasites, we treated mice infected with Plasmodium berghei NYU-2 with chloroquine intraperitoneally in doses ranging from 0.3 to 3 micromol per 25 g mouse. Preparations of infected erythrocytes (normalized to represent 1000 parasites per 1000 erythrocytes) hydrolyzed 1200 nmol of leucine-p nitroanilide per minute per milliliter of packed erythrocytes, which was 10x more than that of uninfected preparations. The activity in infected preparations was distinguished by resistance to ferriprotoporphyrin IX and puromycin and susceptibility to inhibition by ethanol and Tris. Chloroquine treatment caused the activity in unwashed membrane ghosts of infected preparations to decrease by 50% despite an increase in total activity. Concomitantly, hemoglobin in washed membrane ghosts increased. Electron microscopy revealed that the hemoglobin was retained in endocytic vesicles. Chloroquine-induced redistribution of a neutral aminopeptidase may be the cause of hemoglobin accumulation in endocytic vesicles of malaria parasites. PMID- 12573291 TI - Comparative studies of active site-ligand interactions among various recombinant constructs of human beta-amyloid precursor protein cleaving enzyme. AB - Amyloid precursor protein (APP) cleaving enzyme (BACE) is the enzyme responsible for beta-site cleavage of APP, leading to the formation of the amyloid-beta peptide that is thought to be pathogenic in Alzheimer's disease (AD). Hence, BACE is an attractive pharmacological target, and numerous research groups have begun searching for potent and selective inhibitors of this enzyme as a potential mechanism for therapeutic intervention in AD. The mature enzyme is composed of a globular catalytic domain that is N-linked glycosylated in mammalian cells, a single transmembrane helix that anchors the enzyme to an intracellular membrane, and a short C-terminal domain that extends outside the phospholipid bilayer of the membrane. Here we have compared the substrate and active site-directed inhibitor binding properties of several recombinant constructs of human BACE. The constructs studied here address the importance of catalytic domain glycosylation state, inclusion of domains other than the catalytic domain, and incorporation into a membrane bilayer on the interactions of the enzyme active site with peptidic ligands. We find no significant differences in ligand binding properties among these various constructs. These data demonstrate that the nonglycosylated, soluble catalytic domain of BACE faithfully reflects the ligand binding properties of the full-length mature enzyme in its natural membrane environment. Thus, the use of the nonglycosylated, soluble catalytic domain of BACE is appropriate for studies aimed at understanding the determinants of ligand recognition by the enzyme active site. PMID- 12573292 TI - Structural changes and facilitated association of tropoelastin. AB - Circular dichroism studies of tropoelastin secondary structure show 4+/-1% alpha helix in aqueous solutions. This is in contrast to the substantially higher amounts (up to 23+/-7%) of alpha-helix predicted by computer algorithms, which propose that regions of alpha-helix are limited to the alanine-rich cross-linking domains. Through the addition of trifluoroethanol, the amount of alpha-helix increased to 17+/-1%, equivalent to that expected on the basis of primary structure. The physiological ability of the protein to coacervate and the critical concentration of monomer required for coacervation were unaffected by levels of alpha-helix. However, the temperature required for coacervation decreased linearly with increasing alpha-helical structure, which correlates with the participation of alpha-helices in association. We propose that the alanine rich cross-linking domains exist as nascent helices in tropoelastin in aqueous solution. We further suggest a novel mechanism for coacervation whereby formation of alpha-helices and subsequent helical side chain interactions limit the conformational flexibility of the polypeptide, to facilitate associations between hydrophobic domains during elastogenesis. PMID- 12573293 TI - Involvement of cytokines in eating disorders: a critical review of the human literature. AB - A number of findings from clinical and animal studies indicate that pro inflammatory cytokines may play roles in eating disorders. The measurement of pro inflammatory cytokines (IL-1, IL-6, TNFalpha), which are known to decrease food intake, provides highly variable data from which firm conclusions cannot be drawn. In most of the longitudinal studies where pro-inflammatory cytokines have been shown to be impaired in anorexia or bulimia nervosa, a return to normal values was observed after renutrition. However these findings do not exclude the possibility that pro-inflammatory cytokines might be overproduced in specific brain areas and act locally without concomitantly increased serum or immune production. It was also pointed out that the production of the major type-1 cytokines (especially IL-2) was depressed in anorexia nervosa. It remains unclear whether this is due to undernutrition or to a specific underlying cause common to eating disorders. The impaired cytokine profile observed in eating disorders could be related to several factors including impaired nutrition, psychopathological and neuroendocrine factors. More particular attention should be devoted to the deregulation of the anti/pro-inflammatory balance. Deregulation of the cytokine network may be responsible for medical complications in eating disorder patients who are afflicted with chronic underweight. PMID- 12573294 TI - The number of male pups within a litter of NMRI mice is associated with the dam's food preferences late in pregnancy. AB - Pregnancy related differences in food preferences and food intake are well recognized in animals and humans. Changes in maternal hormonal profiles are supposed to be implicated in such a phenomenon. Since fetal sex hormones are excreted in their dams' bloodstream, they can affect the behaviour of mothers during gestation. We investigated whether there was a relationship between the number of male pups within the litter and food preferences in pregnant mice. In the first experiment we found that, parcelling out the effect of litter size, the number of male fetuses was positively correlated to maternal testosterone levels on pregnancy day (PD)18. In the second experiment we showed an association between the number of males and the consumption of a 0.25% saccharin solution on PD18, controlling for litter size and baseline (before pregnancy) saccharin intake. In the third experiment, mice on PD18 were placed in a test cage and underwent a 1-h binary choice test between two unknown unpalatable diets, a high fat diet (HFD) and a sweet mesh diet (HSD). Results revealed that the number of males was negatively associated with the preference for the HFD at birth. These results are consistent with the hypothesis that the behavioural profile of dams can be associated with fetal gender composition. PMID- 12573295 TI - Impact of oral contraceptive use on glucocorticoid sensitivity of pro inflammatory cytokine production after psychosocial stress. AB - We previously reported that women using oral contraceptives (OC) show blunted free cortisol responses to psychosocial stress compared to medication-free women. Low cortisol responses to stress have been shown to be associated with increased susceptibilities to chronic inflammatory and autoimmune processes in animal models and certain human diseases.To address the question if the blunted free cortisol response of OC users may be compensated at the level of the target tissue, we measured hypothalamus-pituitary-adrenal (HPA) axis activation and glucocorticoid (GC) sensitivity of pro-inflammatory cytokine production after psychosocial stress in 14 women using OC and 11 women in the luteal phase of the menstrual cycle. All subjects were exposed to the psychosocial stress paradigm 'Trier Social Stress Test' (TSST). Free cortisol was measured repeatedly before and after stress. GC sensitivity was assessed by dexamethasone (DEX) inhibition of lipopolysaccharide (LPS) stimulated production of interleukin-6 (IL-6) in whole blood, immediately before, as well as 10 and 60 min after the stress test. As expected, the stress test induced significant increases in free cortisol in luteal phase women, while OC users showed blunted responses (F=3.31;p<0.05). GC sensitivity showed different response patterns; In luteal phase women a slight but not significant decrease was observed throughout the experiment. In contrast, women using OC showed a significant increase in GC sensitivity after stress (F=3.559;p<0.05). These results show, that an increase in GC sensitivity of pro inflammatory cytokine production may at least in part compensate the low cortisol levels seen in OC users after stress. This could be one mechanism to protect women using OC medication from chronic inflammatory and autoimmune diseases. PMID- 12573296 TI - Salivary testosterone is related to both handedness and degree of linguistic lateralization in normal women. AB - The aim of this study was to test the hypothesis that individual differences in testosterone (T) are associated with different patterns of linguistic lateralization and hand preference. Twenty left-handed (LH) and 19 right-handed (RH) women filled in a handedness questionnaire and performed a consonant-vowel dichotic listening test (DL-CV). Salivary T was measured by radioimmunoassay (RIA). LH women showed significantly lower mean salivary T than RH women. T levels were negatively correlated with the absolute value of the DL laterality index. Subjects with right ear advantage (REA) were classified into strongly and weakly lateralized following Wexler et al. method (Brain Lang. 13 (1981) 13). When taking into account hand preference, a pattern emerged in that RH-strongly lateralized and LH-weakly lateralized women showed similar T levels. The lowest level appeared for LH-strongly lateralized women and the highest for RH-weakly lateralized women, being significantly different from each other. The results are discussed in terms of several theories that have proposed a link between testosterone and cerebral lateralization. PMID- 12573297 TI - Sexual orientation and the 2nd to 4th finger length ratio: evidence for organising effects of sex hormones or developmental instability? AB - It has been proposed that human sexual orientation is influenced by prenatal sex hormones. Some evidence examining putative somatic markers of prenatal sex hormones supports this assumption. An alternative suggestion has been that homosexuality may be due to general developmental disruptions independent of hormonal effects. This study investigated the ratio of the 2nd to 4th finger digits (the 2D:4D ratio), a measure often ascribed to the organisational actions of prenatal androgens, and the fluctuating asymmetry (FA-a measure of general developmental disruption) of these features, in a sample of 240 healthy, right handed and exclusively heterosexual and homosexual males and females (N=60 per group). Homosexual males and females showed significantly lower 2D:4D ratios in comparison to heterosexuals, but sexual orientation did not relate to any measures of FA. The evidence may suggest that homosexual males and females have been exposed to non-disruptive, but elevated levels of androgens in utero. However, these data also draw attention to difficulties in the interpretation of results when somatic features are employed as biological markers of prenatal hormonal influences. PMID- 12573298 TI - Thyroid function tests and neurocognitive functioning in children referred for attention deficit/hyperactivity disorder. AB - BACKGROUND: Thyroid abnormalities have been associated with attention deficit/hyperactivity disorder (ADHD) and with other childhood psychiatric disorders. The goal of this study was to determine the relationships between thyroid hormone concentrations, neurocognitive functioning, and psychiatric diagnosis in children. METHODS: Free thyroxine index (FT4I) and thyroid stimulating hormone (TSH) were obtained, along with diagnostic and descriptive information for 338 children referred to a clinic specializing in learning and behavior problems. RESULTS: Thyroid abnormalities were uncommon in children referred for ADHD. After excluding children with thyroid disease, there was a greater proportion with low concentrations of normal FT4I for ADHD-Predominantly Inattentive type (ratio=7.0), but not for ADHD-Combined Type. High concentrations of normal FT4I were associated with mood lability, preoccupations, and lower ratings of attention problems. Thyroxine concentrations within the normal range were differentially associated with ADHD-Combined Type compared to ADHD Predominantly Inattentive, mood disorders, and pervasive developmental disorders. CONCLUSION: Thyroxine concentrations were associated with mood symptoms and unusual behaviors, and were less strongly related to attentional functioning. Thyroxine concentrations were not related to hyperactivity. PMID- 12573299 TI - Neuroendocrine and behavioral effects of high-dose anabolic steroid administration in male normal volunteers. AB - OBJECTIVE: Despite widespread abuse of anabolic-androgenic steroids (AAS), the endocrine effects of supraphysiologic doses of these compounds remain unclear. We administered the AAS methyltestosterone (MT) to 20 normal volunteers in an in patient setting, examined its effects on levels of pituitary-gonadal, -thyroid, and -adrenal hormones, and examined potential relationships between endocrine changes and MT-induced psychological symptoms. METHOD: Subjects received MT (three days of 40 mg/day, then three days of 240 mg/day) or placebo in a fixed sequence with neither subjects nor raters aware of order. Samples were obtained at the ends of the baseline, high-dose MT and withdrawal phases. Potential relationships between hormonal changes and visual analog scale measured mood changes were examined. RESULTS: Significant decreases in plasma levels of gonadotropins, gonadal steroids, sex hormone binding globulin, free T3 and T4, and thyroid binding globulin (Bonferroni t, p<0.01 for each) were seen during high-dose MT; free thyroxine and TSH increased during high-dose MT, with TSH increases reaching significance during withdrawal. No significant changes in pituitary-adrenal hormones were observed. Changes in free thyroxine significantly correlated with changes in aggressiveness (anger, violent feelings, irritability) (r=0.5,p=0.02) and changes in total testosterone correlated significantly with changes in cognitive cluster symptoms (forgetfulness, distractibility) (r=0.52,p=0.02). Hormonal changes did not correlate with plasma MT levels. CONCLUSIONS: Acute high-dose MT administration acutely suppresses the reproductive axis and significantly impacts thyroid axis balance without a consistent effect on pituitary-adrenal hormones. Mood and behavioral effects observed during AAS use may in part reflect secondary hormonal changes. PMID- 12573300 TI - Effects of acute and chronic administration of selective monoamine re-uptake inhibitors in the rat forced swim test. AB - The rat forced swim test (FST) is a model that is used extensively as a screening test for antidepressant activity. It has previously been reported that thorough analysis of behaviour in this model reveals two distinct types of active response - climbing and swimming - and that these are separately evoked by re-uptake inhibitors selective for noradrenaline (NA) and serotonin (5-HT), respectively. In the present study, utilising re-uptake inhibitors selective for NA, talsupram, and 5-HT, 5-chloro-1-(3-dimethylaminopropyl)-1-(4-fluorophenyl)- phthalan (Lu 10 134-C), we examined if this scoring technique could detect the antidepressant potential of a selective serotonin re-uptake inhibitor (SSRI), and whether re uptake inhibitors selective for distinct monoamine systems induce exclusive behavioural responses. We also analysed if chronic antidepressant administration for three weeks was more effective than acute treatment. We found Lu 10-134-C (40 mg/kg; PO) to be behaviourally active in this paradigm. Although treatment with talsupram (40 mg/kg; PO) resulted solely in climbing behaviour, Lu 10-134-C induced both climbing and swimming behaviour. However, chronic pre-treatment with either re-uptake inhibitor (20 mg/kg; twice daily; PO) failed to augment the response observed with acute treatment. Similarly, chronic administration of either compound was without effect on the basal, or stress-induced, serum corticosterone concentrations or anterior pituitary (AP) preproopiomelanocorticotropin (POMC) mRNA expression. These results suggest that selective monoamine re-uptake inhibition produces distinct, but not necessarily exclusive, behavioural responses in the forced swim test. PMID- 12573301 TI - Bicuculline seizure susceptibility and nigral GABAA alpha1 receptor mRNA is altered in adult prenatally morphine-exposed females. AB - Prenatal morphine exposure (5-10 mg/kg twice daily on gestation days 11-18) can adversely affect neurological development, including seizure susceptibility. The present study examines the effects of prenatal morphine exposure on seizure susceptibility to the GABA antagonist and convulsant bicuculline and GABA(A) alpha(1) receptor mRNA in the substantia nigra (SN) of female rats. The results demonstrate that prenatally morphine-exposed ovariectomized (OVX) females and OVX females with estradiol benzoate (EB) replacement have an increased latency to seizure onset compared to controls. In addition, prenatal morphine exposure decreases the area covered by grains of GABA(A) alpha(1) receptor mRNA in the anterior SN in both OVX and EB+progesterone (P)-treated groups, and decreases the number of GABA(A) alpha(1) receptor mRNA-labeled cells/field in EB females. Furthermore, prenatally morphine- and saline-exposed EB and EB+P females had decreased GABA(A) alpha(1) receptor mRNA-labeled cells/field in the anterior SN compared to OVX animals of the same prenatal exposure. These results demonstrate that the long term effects of prenatal morphine exposure in female rats is dependent on their hormonal status, and suggest that seizure susceptibility may be altered via neuropharmacological changes in the GABA system in the SN. PMID- 12573302 TI - Anticipatory cortisol, testosterone and psychological responses to judo competition in young men. AB - This study compares the anticipatory hormonal and psychological responses of 17 male judo players to an official competition with the data obtained during eight resting sessions carried out at the same time of day, throughout an entire sports season. Testosterone (T) and cortisol (C) levels were determined 1 h and 30 min before competition, and mood, anxiety and expectancies were also evaluated. C levels and anxiety scores were concurrently higher before the contest than in resting conditions; however, non-significant correlations between them were found. The anticipatory T response was not significant for the whole group. However, one group of subjects did display T increases, higher C levels, and higher motivation to win scores than the other group. Furthermore, this group also obtained a better outcome. Thus, this hormonal pattern and its relationships with psychological variables suggest an adaptive psychobiological response to a competition. Results are discussed in the context that neuroendocrine response to competition is associated with cognitive appraisal. PMID- 12573303 TI - The combined dexamethasone-CRH test before and after repetitive transcranial magnetic stimulation (rTMS) in major depression. AB - BACKGROUND: Hypothalamic-pituitary-adrenocortical (HPA) dysregulation assessed by the combined dexamethasone corticotropin releasing hormone test (DEX/CRH test) has been demonstrated to normalize after successful antidepressant pharmacotherapy. Here, we investigated whether repetitive transcranial magnetic stimulation (rTMS) also leads to a normalization of HPA system activity in depressed patients. METHODS: Thirty-seven medication free patients suffering from a major depressive episode (DSM-IV) underwent a DEX/CRH test before and after 13 daily sessions of left prefrontal rTMS in an open trial. RESULTS: There was an overshoot of CRH-induced cortisol release that was not affected by rTMS treatment. Postdexamethasone cortisol levels prior to CRH challenge decreased in responders after rTMS treatment, whereas no change of CRH-induced adrenocorticotropic hormone (ACTH) and cortisol release in responders or nonresponders was observed. CONCLUSIONS: The persisting HPA system hyperactivity after rTMS suggests a high risk for relapse and therefore argues for an immediate maintenance therapy in patients responding to this treatment. PMID- 12573304 TI - Intrauterine position has long-term influence on brain mu-opioid receptor density and behaviour in mice. AB - In multiparous rodents, a naturally occurring variation in degree of exposure to sex steroids during the prenatal phase of sexual differentiation derives from the in-utero proximity to opposite sex foetuses. So far, the studies on intrauterine position (IUP) phenomenon have mostly focused on traits relating to reproduction and behaviour, while its influence on neurochemical substrates and pharmacological response has been largely unexplored. We investigated possible variations in the function and the profile of expression of the mu-opioid receptor system in three groups of adult mice from known IUP: 2M mice (located between two males), 0M (between two females), and 1M (between a male and a female). Autoradiographic study revealed in female mice that proximity to at least a male in utero (1M and 2M position) resulted associated at adulthood with an increased density of midbrain mu-opioid receptors. Behavioural observations were conducted following injection with the specific mu-opioid agonist Fentanyl (at 0, 0.01 or 0.05 mg/kg IP). A drug-conditioned place preference test confirmed that 1M and 2M subjects were also more sensitive to the rewarding effects of the drug, since mice spent significantly more time in the drug-paired compartment than 0M subjects. In a hot-plate test, 2M subjects showed levels of drug-induced analgesia that were much higher than other IUP groups. No reliable differences were observed between the IUP groups for locomotor activity upon drug treatment. Overall, these data indicate for the first time that the organisation of the mu opioid receptor system in the brain, as well as a differential vulnerability to abuse of opiate drugs can be modulated by epigenetic variables such as the prenatal in utero contiguity to male foetuses. PMID- 12573305 TI - On the role of menopause for sleep-endocrine alterations associated with major depression. AB - Aging and menopause are associated with alterations of the sleep EEG, while age related changes of the hypothalamo-pituitary-adrenal (HPA) axis remain controversial. Major depression is also associated with typical sleep-endocrine changes, including enhanced activity of the HPA axis, while an influence of age and gender on these alterations is less clear. To test the hypothesis that after menopause sleep-endocrine alterations associated with major depression are accentuated, we examined the sleep EEG and nocturnal hormone secretion (ACTH, cortisol, GH, estradiol, LH, FSH, and leptin) in 16 drug-free female patients, mostly with the first episode of a major depressive disorder (seven pre- and nine postmenopausal subjects) and 19 female controls (10 subjects in the early follicular phase and nine postmenopausal subjects). Nocturnal cortisol secretion was increased in postmenopausal patients with depression, while a decrease was noted in postmenopausal controls. Sleep alterations typically associated with depression, namely a reduction in sleep continuity and slow wave sleep (SWS) and an increase in REM density, were prominent in post- but not in premenopausal patients. An inverse correlation was noted between the decline in SWS and sleep continuity and FSH secretion in patients with depression, suggesting a role of menopause for these sleep-endocrine alterations typically associated with major depression. In contrast, in premenopausal patients we noted primarily a shift in SWS and delta-EEG activity from the first to the second non-REM period, which was not related to age or hormone secretion. Though the relatively small number of subjects per group precludes a definitive conclusion, our data open up the possibility that the sleep-endocrine changes typically associated with major depression are most prominent in postmenopausal patients. Whether the predominant alteration of the distribution of SWS and delta EEG activity in younger patients with a first episode of major depression has a predictive value for the future course of the disease remains to be investigated. PMID- 12573306 TI - Effects of hormone replacement therapy on perceptual and cognitive event-related potentials in menopausal insomnia. AB - The influence of a combined estrogen-progestin regimen (Climodien, Lafamme) on auditory event-related potentials (ERPs) was investigated in a double-blind, placebo-controlled, comparative, randomized 3-arm trial phase (Climodien 2/3=estradiol valerate 2 mg+the progestin dienogest 3 mg, EV=estradiol valerate 2 mg, and placebo), followed by an open-label phase in which all patients received Climodien 2/2 (estradiol valerate 2 mg+dienogest 2 mg). Both the double-blind and the open-label phase lasted 2 months. ERPs were recorded from 19 EEG leads in a two-tone odd-ball paradigm in 49 patients aged between 46 and 67 yr with the diagnosis of insomnia (G 47.0) related to postmenopausal syndrome (N 95.1). Climodien reduced standard N1 and target P300 latencies as compared to placebo, while EV did not affect N1 latency but similarly reduced P300 latency. Climodien increased N1, P2 and P300 amplitudes dose-dependently, predominantly at frontal leads. Estrogen alone had only minor effects on ERP amplitudes. The shortening of standard N1 latency and enhancement of N1 and P2 amplitudes indicates a positive effect of Climodien on perceptual processing, most likely due to vigilance improvements also observed in EEG mapping. Concerning target P300, it seems that estradiol is responsible for the improvement in stimulus evaluation time, as reflected by the shortening of the peak latency, while dienogest seems to account for the improvement in cognitive information processing capacity, whereby 3 mg induced a more pronounced augmentation of P300 amplitudes than 2 mg. Based on the spatial distribution of this increase, it can be speculated that Climodien mainly affects the more frontally distributed P3a subcomponent, which is associated with attention and orientation. Furthermore, the observed changes in ERP-components are consistent with recent studies showing significant positive effects of hormone replacement therapy on cholinergic functions. Thus, Climodien seems to be of interest in preventing cognitive decline and treating cognitive disorders in postmenopausal women. Indeed, there is increasing evidence of beneficial effects of estrogen in dementia. Our present findings suggest that the estrogen effects may be augmented by dienogest. PMID- 12573307 TI - Serotonin transporter gene polymorphisms and platelet [3H] paroxetine binding in premenstrual dysphoria. AB - The purpose of this study was to investigate if women with premenstrual dysphoria differ from controls with respect to the number of platelet serotonin transporters, and with respect to three polymorphisms in the gene coding for the serotonin transporter: a 44 base pair insertion/deletion in the promoter region, a variable number of tandem repeats in the second intron, and a single nucleotide polymorphism in the 3' untranslated region. Also, the possible relationship between the three polymorphisms and platelet serotonin transporter density was analyzed. The density of platelet [(3)H]paroxetine binding sites was significantly lower in women with premenstrual dysphoria than in controls, but patients and controls did not differ with respect to allele or genotype frequency for any of the three polymorphisms examined. A significant association between the number of platelet serotonin transporters and the promoter polymorphism was observed, subjects being homozygous for the short (deletion) variant having higher platelet serotonin transporter density than subjects carrying the long (insertion) allele. The results support the assumption that serotonin-related psychiatric disorders-such as premenstrual dysphoria-may be associated with a reduction in platelet [(3)H]paroxetine binding, but argue against the notion that this reduction is due to certain variants of the serotonin transporter gene being more common in patients than in controls. PMID- 12573308 TI - Daily psychosocial stressors and cyclic response patterns in urine cortisol and neopterin in a patient with systemic lupus erythematosus. AB - This study investigated the complex biochemical responses to personally meaningful everyday stressors in a patient with systemic lupus erythematosus (SLE). For this purpose, a 52 year-old woman with SLE collected her entire urine for 56 days on a 12-h basis for the determination of cortisol as well as neopterin, a cellular immune parameter. Additionally, using questionnaires, daily notes and interviews, extensive psychosocial and psychological time-series data were collected every 12 h. Cross-correlational analyses of the resulting time series revealed that stressful incidents were associated with cyclic fluctuations in both urine cortisol and urine neopterin. Specifically, whenever the patient anticipated a moderately stressful incident, urine cortisol initially increased 24 h before the incident and then decreased 12 h before the incident. Moderate stressors not anticipated by the patient were associated with an initial increase 24 h following the incident and then with a decrease after a total of 36 h. Moreover, stressors having to do with the patient's extramarital relationship were followed initially by a decrease in urine neopterin after 36 h and then by an increase after a total of 60 h. Our findings indicate that when investigating the relationship between psychosocial stressors and biochemical activity in SLE, appropriate consideration of the data's dynamic nature may be necessary to avoid flawed conclusions. PMID- 12573313 TI - Thermodynamic and kinetic investigations of uranium adsorption on amberlite IR 118H resin. AB - The adsorption behavior of uranium(VI) on a strongly acidic cation exchanger amberlite IR-118H has been studied as a function of the solution concentration and temperature. Results have been analyzed by Langmuir and Dubinin-Radushkevich (D-R) adsorption isotherms. The mean energy of adsorption 7.14kJmol(-1) was calculated from the D-R adsorption isotherm. In order to understand the significance of the diffusion mechanisms and to accurately estimate the diffusivities inside the adsorbent particles, a model for uranium(VI) adsorption on amberlite IR-118H was used. The rate constants have been calculated for 293, 313 and 333K using Lagergren equation and the activation energy (E(a)) was derived using the Arhenius equation. The thermodynamic quantities for the process of adsorption have been estimated by plotting Ln K(D) versus 1/T. The DeltaH(o) and DeltaG(o) values of uranium(VI) adsorption on amberlite IR-118H show endothermic heat of adsorption; higher temperatures favor the process. PMID- 12573314 TI - Transfer of Eu (III) associated with polymaleic acid to Bacillus subtilis. AB - The aim of this study is to contribute to the understanding of the distribution of Eu(III) between dissolved organic matter and microorganisms, and to investigate the effect of competitive ions such as Ca(+2) on adsorption properties. Polymaleic acid (PMA), is used as synthetic organic matter, having similar properties as natural fulvic acid, and Bacillus subtilis is chosen as microorganism. A double labeling method was used: [14C]MPA and 152Eu to quantify the behavior of the various components. Preliminary experiments showed that the adsorption of polymaleic acid onto Bacillus subtilis was negligible at pH=5 in 0.15mol/l of NaCl. In the absence of Ca(+2), the transfer of Eu(III) from PMA to B. subtilis could be described by a simple empirical model based on data obtained from sorption isotherms on the reference systems Eu(III)/PMA and Eu(III)/B. subtilis. In the presence of Ca(+2), the transfer was increased. The hypothesis that Ca(+2) ions acted as a bridging agent between PMA and the bacteria was proposed. PMID- 12573315 TI - Application of activation methods on the Dubna experimental transmutation set ups. AB - High spallation neutron fluxes were produced by irradiating massive heavy targets with proton beams in the GeV range. The experiments were performed at the Dubna High Energy Laboratory using the nuclotron accelerator. Two different experimental set-ups were used to produce neutron spectra convenient for transmutation of radioactive waste by (n,x) reactions. By a theoretical analysis neutron spectra can be reproduced from activation measurements. Thermal epithermal and fast-super-fast neutron fluxes were estimated using the 197Au, 238U (n,gamma) and (n,2n) reactions, respectively. Depleted uranium transmutation rates were also studied in both experiments. PMID- 12573316 TI - An X-ray micro-tomography system optimised for the low-dose study of living organisms. AB - An X-ray micro-tomography system has been designed that is dedicated to the low dose imaging of radiation sensitive living organisms and has been used to image the early development of the first few days of plant development immediately after germination. The system is based on third-generation X-ray micro-tomography system and consists of an X-ray tube, two-dimensional X-ray detector and a mechanical sample manipulation stage. The X-ray source is a 50kVp X-ray tube with a silver target with a filter to centre the X-ray spectrum on 22keV.A 100mm diameter X-ray image intensifier (XRII) is used to collect the two-dimensional projection images. The rotation tomography table incorporates a linear translation mechanism to eliminate ring artefact that is commonly associated with third-generation tomography systems. Developing maize seeds (Triticum aestivum) have been imaged using the system with a cubic voxel linear dimension of 100 microm, over a diameter of 25mm and the root lengths and volumes measured. The X ray dose to the plants was also assessed and found to have no effect on the plant root development. PMID- 12573317 TI - Synthesis of N,N-dimethyl-2-(2-amino-4-[18F]fluorophenylthio)benzylamine as a serotonin transporter imaging agent. AB - Two 403U76 analogs, N,N-dimethyl-2-(2-nitro-4-bromophenylthio)benzylamine (4) and N,N-dimethyl-2-(2,4-dinitrophenylthio)benzylamine (8) were prepared in multi steps synthesis as precursors for the synthesis of a new serotonin transporter imaging agent, N,N-dimethyl-2-(2-amino-4-[18F]fluorophenylthio)benzylamine (12a). Reaction of 2,5-dibromonitrobenzene (1) with 2-thio-N,N-dimethylbenzamide gave N N-dimethyl-2-(2-nitro-4-bromophenylthio)benzamide (3). N,N-Dimethyl-2-(2,4 dinitrophenylthio)benzamide (6) was synthesized similarly from the reaction of 2 bromo-1,5-dinitrobenzene (2) with 2-thio-N,N-dimethylbenzamide. Reduction of 3 and 6 with BH(3)/THF gave benzylamines 4 and 8 along with their amine boranes 5 and 7. Nucleophilic substitution of 4 or 8 with K[18F]/Kryptofix 2.2.2 in DMSO at 120 degrees C followed by reduction with NaBH(4)- Cu(OAc)(2) in EtOH at 78 degrees C and purification with HPLC gave compound 12a in approximately 5-10% yield in a synthesis time of 150min from EOB. A preliminary biodistribution study in rats showed that the uptake of compound 12a in rat brain was high (approximately 1%/g) and the ratio of the uptake of compound 12a in hypothalamus (a serotonin transporter-rich area) and cerebellum (a serotonin-transporter devoid area) was 6/1 at 1h post-injection. These results suggest that compound 12a may be a potential new serotonin transporter PET imaging agent. PMID- 12573318 TI - Rapid screening of 2-[18F]-fluoro-2-deoxy-D-glucose infusions for volatile organic compound contaminants by solid phase microextraction with gas chromatography-selective ion monitoring mass spectrometry (SPME-GC-SIMMS). AB - A method compatible with radioactive samples, capable of detecting trace volatile components in a sample volume of ca. 1cm(3) of 2-[18F]-fluoro-2-deoxy-D-glucose solution is described. The approach, based on solid phase micro-extraction gas chromatography-mass spectrometry with a carboxen/polydimethylsiloxane based fibre, was optimised with respect to extraction time (10 min), extraction temperature (60 degrees C) and phase volume ratio (1). The analysis time, including extraction, was less than 20 min with linear responses for acetonitrile and ethanol over the ranges: 0.09-80 microg cm(-3) (22 degrees C, acetonitrile) and 0.78-79 microg cm(-3) (22 degrees C, ethanol). The detection limits were estimated to be ca. 0.78 microg cm(-3) for ethanol and 0.09 microg cm(-3) for acetonitrile. Stability studies indicated analyte losses of up to 75% over 24h and analysis of aged 2-[18F]FDG samples showed that levels of ethanol and acetonitrile were not less than 100 microg cm(-3), indicative of levels substantially greater than this in the original infusions given to human subjects. PMID- 12573319 TI - Synthesis and evaluation of a 18F-labelled recombinant annexin-V derivative, for identification and quantification of apoptotic cells with PET. AB - In this report, we describe the synthesis of 4-[18F]-fluorobenzoyl-annexin V (4 [18F]FBA). In a four-step procedure, 4-[18F]FBA was synthesised with a microcomputer controlled, automated module within 90min. The radiochemical yield was in the range of 15-20% (corrected for decay) with a specific activity of more than 35GBq/micromol. The specific binding was confirmed by studies of 4-[18F]FBA with phosphatidylserine-containing liposomes. The biological activity of 4 [18F]FBA was verified by measuring its binding to Jurkat T-cell lymphoblasts after induction of apoptosis as compared to control cells without apoptosis. 4 [18F]FBA seems to be a suited tracer to measure apoptotic activity in vivo. PMID- 12573320 TI - Radiosynthesis of ML03, a novel positron emission tomography biomarker for targeting epidermal growth factor receptor via the labeling synthon: [11C]acryloyl chloride. AB - An automated procedure for the radiosynthesis of the labeling synthon [11C]acryloyl chloride was developed and applied for labeling several N-acryl amides with carbon-11. [11C]-6-acrylamido-4-(3,4-dichloro-6 fluoroanilino)quinazoline (ML03), a novel PET biomarker targeting the epidermal growth factor receptor tyrosine kinase (EGFr-TK) in cancer, was successfully prepared using this labeled synthon in a fully automated manner. Two other potential anticancer drugs were also labeled using the developed methodology. The potency of ML03 to inhibit autophosphorylation of EGFr-TK was evaluated by an ELISA assay indicating a low IC(50) of 0.037nM. PMID- 12573321 TI - Pharmacokinetics and radiation dosimetry estimation of O-(2-[18F]fluoroethyl)-L tyrosine as oncologic PET tracer. AB - An easy-to-automate synthetic procedure and the kinetics and radiation dosimetry of O-(2-[18F]fluoroethyl)-L-tyrosine (FET), a recently developed amino acid tracer with potential applications in tumor imaging with PET, are described. FET was prepared in high radiochemical yield, 20-25% with no decay correction, and radiochemical purity of more than 95% in less than 60min synthesis time by a modified two-step procedure and manual operation. The kinetics and radiation dosimetry of FET were evaluated by using mice biodistribution data and the medical internal radiation dosimetry (MIRD) method. The bone (total) was the organ receiving the highest dose, 4.78x10(-3)mGy/MBq, and the brain and the whole body received the lowest dose, 1.6x10(-3)mGy/MBq, respectively. The effective dose was 9.0x10(-3)mSv/MBq. The data show that a 370-MBq (10mCi) injection of FET leads to an estimated effective dose of 3.3mSv and an estimated dose to the whole body of 0.6mGy. The potential radiation risks associated with this study are well within accepted limits. PMID- 12573322 TI - Absolute calibration of 60Co by using sum-peak method and an HPGe detector. AB - The sum-peak method was applied to calibrate four 60Co sources in the range of 25 350kBq. An HPGe-based gamma-ray spectroscopy system was used, and the effects of the angular correlation and pulse pile-up were corrected, and are discussed. The correction for the angular correlation was obtained from a theoretical calculation, while the pulse pile-up effect was corrected using a new extrapolation technique based on the pile-up measurements by varying the shaping time of the amplifier. The determined radioactivities were within 2% deviation from the values reported by the supplier and the associated uncertainties were less than 2%. The observed consistency was obtained under the experimental condition of keeping the total counting rate 75% transmural extent of the left ventricular wall). CMR also identified 100 of the 109 segments (92%) with subendocardial infarction (<50% transmural extent of the left-ventricular wall), whereas SPECT identified only 31 (28%). SPECT and CMR showed high specificity for the detection of infarction (97% and 98%, respectively). In patients, all segments with nearly transmural infarction, as defined by contrast-enhanced CMR, were detected by SPECT. However, of the 181 segments with subendocardial infarction, 85 (47%) were not detected by SPECT. On a per patient basis, six (13%) individuals with subendocardial infarcts visible by CMR had no evidence of infarction by SPECT. INTERPRETATION: SPECT and CMR detect transmural myocardial infarcts at similar rates. However, CMR systematically detects subendocardial infarcts that are missed by SPECT. PMID- 12573374 TI - Folic acid supplements during early pregnancy and likelihood of multiple births: a population-based cohort study. AB - BACKGROUND: Folic acid supplements are recommended for women of childbearing age to prevent neural tube defects in their offspring. Results of some studies, however, suggest an increase in multiple births associated with use of vitamin supplements that contain folic acid during pregnancy. Our aim was to assess this association. METHODS: We used data from a population-based cohort study from which we assessed the occurrence of multiple births in women (n=242015) who had participated in a campaign to prevent neural tube defects with folic acid supplements (400 microg per day) in China. Folic acid use was ascertained before pregnancy outcome was known. We studied the relation between multiple births and any use of folic acid pills before or during early pregnancy; additionally, we investigated mechanisms by which folic acid could potentially affect the occurrence of multiple births by examining pill-taking at three time periods: before ovulation, around the time of fertilisation, and after conception. FINDINGS: 1496 (0.62%) multiple births occurred in a cohort of 242015 women who had registered with the study between October, 1993, and September, 1995, and who had a pregnancy not affected by a birth defect; the rate of multiple births in women who did and did not take folic acid before or during early pregnancy was 0.59% and 0.65%, respectively (rate ratio 0.91; 95% CI 0.82-1.00). INTERPRETATION: Our findings suggest that consumption of folic acid supplements during pregnancy is not associated with an increased occurrence of multiple births. PMID- 12573375 TI - Purpura fulminans. PMID- 12573377 TI - Sudden death following a ventral hernia repair. PMID- 12573376 TI - Sensory fibres expressing capsaicin receptor TRPV1 in patients with rectal hypersensitivity and faecal urgency. AB - BACKGROUND: Faecal urgency and incontinence with rectal hypersensitivity is a distressing, unexplained disorder that is inadequately treated. We aimed to determine whether expression of the heat and capsaicin receptor vanilloid receptor 1 (TRPV1 or VR1) was changed in rectal sensory fibres, and to correlate nerve fibre density with sensory abnormalities. METHODS: We compared full thickness rectal biopsy samples from nine patients with physiologically characterised rectal hypersensitivity with tissue samples from 12 controls. Sensory thresholds to rectal balloon distension and heating the rectal mucosa were measured before biopsy. We assessed specimens with immunohistochemistry and image analysis using specific antibodies to TRPV1; nerve growth factor (NGF) receptor tyrosine kinase A; glial cell line-derived neurotrophic factor (GDNF); neuropeptides calcitonin gene-related peptide (CGRP) and substance P; the related vanilloid receptor-like protein (VRL) 2; glial markers S-100 and glial fibrillary acid protein (GFAP); and the nerve structural marker peripherin. FINDINGS: In rectal hypersensitivity, nerve fibres immunoreactive to TRPV1 were increased in muscle, submucosal, and mucosal layers: in the mucosal layer, the median% area positive was 0.44 (range 0.30-0.59) in patients who were hypersensitive and 0.11 (0.00-0.21) in controls (p=0.0005). The numbers of peripherin-positive fibres also increased in the mucosal layer (hypersensitive 3.00 [1.80-6.50], controls 1.20 [0.39-2.10]: (p=0.0002). The increase in TRVP1 correlated significantly with the decrease in rectal heat (p=0.03) and the distension (p=0.02) sensory thresholds. The thresholds for heat and distension were also significantly correlated (p=0.0028). Expression of nerve fibres positive for GDNF (p=0.001) and tyrosine kinase A (p=0.002) was also increased, as were cell bodies of the submucosal ganglia immunoreactive to CGRP (p=0.0009). INTERPRETATION: Faecal urgency and rectal hypersensitivity could result from increased numbers of polymodal sensory nerve fibres expressing TRPV1. The triggering factor or factors remain uncertain, but drugs that target nerve terminals that express this receptor, such as topical resiniferatoxin, deserve consideration. PMID- 12573378 TI - Rapidly reversible dementia. PMID- 12573379 TI - Association between telomere length in blood and mortality in people aged 60 years or older. AB - During normal ageing, the gradual loss of telomeric DNA in dividing somatic cells can contribute to replicative senescence, apoptosis, or neoplastic transformation. In the genetic disorder dyskeratosis congenita, telomere shortening is accelerated, and patients have premature onset of many age-related diseases and early death. We aimed to assess an association between telomere length and mortality in 143 normal unrelated individuals over the age of 60 years. Those with shorter telomeres in blood DNA had poorer survival, attributable in part to a 3.18-fold higher mortality rate from heart disease (95% CI 1(.)36-7.45, p=0.0079), and an 8.54-fold higher mortality rate from infectious disease (1.52-47.9, p=0.015). These results lend support to the hypothesis that telomere shortening in human beings contributes to mortality in many age-related diseases. PMID- 12573380 TI - Effectiveness of vaccination for Haemophilus influenzae type b. PMID- 12573381 TI - Large-scale evidence that the cardiotoxicity of smoking is not significantly modified by the apolipoprotein E epsilon2/epsilon3/epsilon4 genotype. AB - Results from two small studies, involving a total of only 174 cases, have suggested that the increased risk of coronary heart disease conferred by cigarette smoking is substantially affected by genotype at the apolipoprotein E (APOE) epsilon2/epsilon3/epsilon4 polymorphism. We have established APOE genotypes in 4484 patients with acute myocardial infarction diagnosed before the age of 55 years for male and 65 years for female patients, and in 5757 controls with no history of cardiovascular disease. On average, the hazard ratio for myocardial infarction was 1.17 (95% CI 1.09-1.25; p<0.00001) per stepwise change from epsilon3/2 to epsilon3/3 to epsilon3/4 genotype. Among individuals in this study with known cigarette smoking status, the hazard ratio for myocardial infarction in smokers versus non-smokers was 4.6 (4.2-5.1). There was, however, no significant difference between the smoker/non-smoker hazard ratios for those with different APOE genotypes (chi2(2)=0.69; p=0.7). When differences in risk between different genotypes are not extreme (as with this APOE polymorphism), reliable assessment of hypothesised gene-environment interactions will often require the study of many thousands of disease cases. PMID- 12573382 TI - South Korea's Jong Wook Lee elected new WHO Director-General. PMID- 12573383 TI - Pascoal Mocumbi--international health advocate. PMID- 12573387 TI - Eating disorders. AB - Eating disorders are an important cause of physical and psychosocial morbidity in adolescent girls and young adult women. They are much less frequent in men. Eating disorders are divided into three diagnostic categories: anorexia nervosa, bulimia nervosa, and the atypical eating disorders. However, the disorders have many features in common and patients frequently move between them, so for the purposes of this Seminar we have adopted a transdiagnostic perspective. The cause of eating disorders is complex and badly understood. There is a genetic predisposition, and certain specific environmental risk factors have been implicated. Research into treatment has focused on bulimia nervosa, and evidence based management of this disorder is possible. A specific form of cognitive behaviour therapy is the most effective treatment, although few patients seem to receive it in practice. Treatment of anorexia nervosa and atypical eating disorders has received remarkably little research attention. PMID- 12573389 TI - Unjust embargo of aid for Haiti. PMID- 12573388 TI - Genes for schizophrenia? Recent findings and their pathophysiological implications. AB - CONTEXT: Schizophrenia is highly heritable, but the genes have remained elusive. Identifying the genes is essential if the pathogenesis and pathophysiology of schizophrenia is finally to be understood, and to give the prospect of more effective treatment. STARTING POINT: H Stefansson and colleagues (Am J Hum Genet 2002; 71: 877-92) showed association of the neuregulin (NRG1) gene with schizophrenia. Other recent papers describe six additional susceptibility genes. Replications are already being reported for some of them. The genes are biologically plausible, and may have convergent effects on glutamatergic and other synapses. We review the evidence for each gene, the possible pathogenic mechanisms, and the implications of the findings. WHERE NEXT? Given earlier failures to replicate apparent breakthroughs, the results should be viewed with caution. Unequivocal replications remain the top priority. The respective contributions of each gene, epistatic effects, and functional interactions between the gene products, all need investigation. Confirmation that any of the genes is a true susceptibility gene for schizophrenia could trigger the same rapid therapeutic progress as has occurred recently in Alzheimer's disease. PMID- 12573390 TI - What is the WMA for? The case of the Israeli Medical Association. PMID- 12573393 TI - The PROSPER trial. PMID- 12573394 TI - The PROSPER trial. PMID- 12573395 TI - The PROSPER trial. PMID- 12573396 TI - The PROSPER trial. PMID- 12573397 TI - The PROSPER trial. PMID- 12573399 TI - Effect of paracetamol poisoning on international normalised ratio. PMID- 12573401 TI - Mortality and indicators of traffic-related air pollution. PMID- 12573403 TI - The ISAT trial. PMID- 12573404 TI - The ISAT trial. PMID- 12573405 TI - The ISAT trial. PMID- 12573406 TI - The ISAT trial. PMID- 12573409 TI - A neglected breakthrough in asthma therapy. PMID- 12573410 TI - Effect of immunisation status on asthma prevalence. PMID- 12573411 TI - Acrylamide food risk. PMID- 12573412 TI - Antiretroviral treatment and research in resource-poor countries. PMID- 12573413 TI - Consumption of arsenic through cooked rice. PMID- 12573414 TI - Antiretroviral treatment and research in resource-poor countries. PMID- 12573415 TI - Risk factors for preterm delivery. PMID- 12573416 TI - Risk factors for preterm delivery. PMID- 12573418 TI - Hazards of widespread use of erythromycin for preterm prelabour rupture of membranes. PMID- 12573419 TI - El Nino and drought in southern Africa. PMID- 12573420 TI - Bridges to Iran. PMID- 12573421 TI - Cause of death among passengers on the Titanic. PMID- 12573423 TI - Rules for romance. PMID- 12573426 TI - Bioterrorism alert. PMID- 12573429 TI - Ras redux: rethinking how and where Ras acts. AB - Ras proteins couple surface receptor stimulation to an ever-expanding array of effector pathways. The integrated activities of these signaling pathways result in a range of biological responses depending on cellular context and conditions. Decoding the various inputs that underlie Ras-induced proliferation and transformation, in particular, holds the promise of more effective cancer therapies. Recent findings have provided new and unexpected insights into the predominant pathways Ras employs to transform human cells and the subcellular platforms from which it can act. PMID- 12573430 TI - Genomic approaches to identification of tumour-susceptibility genes using mouse models. AB - Individual susceptibility to cancer in humans is determined by complex interactions between germline genetic variation and levels of exposure to environmental carcinogens or tumour promoters. Only a small fraction of cancer susceptibility is inherited in a Mendelian manner (high-penetrance familial cancer), and most tumours result from the combined effects of many gene-gene and gene-environment interactions. The sequencing of the mouse genome provides new approaches to one of the most challenging tasks of cancer genetics today. PMID- 12573431 TI - Neurofibromatosis 1: closing the GAP between mice and men. AB - Neurofibromatosis 1 (NF1) is a common genetic condition in which affected individuals are prone to the development of benign and malignant tumors. The NF1 tumor suppressor encodes a protein product, neurofibromin, which functions in part as a negative regulator of RAS. Loss of neurofibromin expression in NF1 associated tumors or Nf1-deficient mouse cells is associated with elevated RAS activity and increased cell proliferation. Despite this straightforward pathophysiologic association between neurofibromin, RAS, and tumorigenesis, recent insights from mouse and Drosophila modeling studies have suggested additional functions for neurofibromin and have implicated Nf1 heterozygosity in tumor formation. Lastly, Nf1 knockout mouse studies have also demonstrated important roles for cooperating genetic changes that accelerate tumorigenesis as well as modifier genes that impact on cancer susceptibility. PMID- 12573432 TI - You Wnt some, you lose some: oncogenes in the Wnt signaling pathway. AB - The highly regulated Wnt signaling cascade plays a decisive role during embryonic patterning and cell-fate determination. The inappropriate expression of Wnt target genes, resulting from deregulation of this pathway, is also implicated in tumorigenesis. Thus, regulation of this pathway is of paramount importance. The Wnt signals are extracellularly regulated by a diverse group of antagonists, cofactors and coreceptors. In the cytoplasm, beta-catenin, a key effector of the Wnt signaling cascade, is highly regulated by a large and fascinating complex of proteins. In the nucleus, activation of target genes is regulated by a complex interplay of activators, repressors and other proteins. Recently, new factors in this pathway have been identified and the interplay and mechanisms of action of key players have been better characterized. Collectively, this represents an important step forward in our understanding of the role of Wnt signaling in development and oncogenesis. PMID- 12573433 TI - Sonic hedgehog in normal and neoplastic proliferation: insight gained from human tumors and animal models. AB - Cancer arises when a cell accumulates multiple genetic changes that allow it to elude the highly regulated balance between proliferation and apoptosis that an organism employs to suppress inappropriate growth. It has become evident that malignant transformation of a cell or group of cells often involves pathways that are active during normal development but are inappropriately regulated in neoplastic proliferation. Signaling via the Sonic hedgehog pathway is critical to vertebrate development and also appears to play an integral role in the initiation and propagation of some tumors of the muscle, skin and nervous system. Analyses of human tumors have revealed mutations in various components of the Sonic hedgehog signaling pathway that appear to result in the activation of this pathway, as inferred by the increased expression of the transcription factor, Gli1. Interestingly, a proportion of the human tumors and most of those arising in mouse models continue to express the normal Patched allele, suggesting the involvement of additional molecular events in the transformation of the haploinsufficient cells. PMID- 12573434 TI - RUNX transcription factors as key targets of TGF-beta superfamily signaling. AB - The Runt domain transcription factors, RUNX1, RUNX2 and RUNX3, are integral components of signaling cascades mediated by both TGF-beta and bone morphogenetic proteins (BMPs) in several important biological systems. RUNX2 functions synergistically with Smad1 and Smad5 to regulate bone-specific genes when BMP induces osteogenesis. RUNX3, which has been mapped to locus 1p36, is a major tumor suppressor of gastric cancer and appears to be an important component of the TGF-beta-induced tumor suppressor pathway. A possible relationship between the TGF-beta-induced tumor-suppressor pathway and a postulated tumor suppressor gene on 1p36 must be examined. PMID- 12573435 TI - The core-binding factor leukemias: lessons learned from murine models. AB - The recent development of murine models of core-binding factor leukemias has provided important insights into the underlying molecular pathology of this common subtype of acute myeloid leukemia. Evidence from these models supports the idea that acute myeloid leukemia 1/core-binding factor beta-subunit (AML1/CBFbeta) has a critical role in the control of the self-renewal capacity of hematopoietic stem cells and their progeny. Moreover, the accumulated data demonstrate that the expression of translocation-encoded AML1 or CBFbeta fusion proteins are insufficient by themselves to induce a full leukemic phenotype. The models that have been developed should prove to be of value for defining the range of mutations that can cooperate with AML1/CBFbeta fusion proteins, and for assessing novel therapies targeted toward the pathways that are altered by the expression of these fusion proteins. PMID- 12573436 TI - The von Hippel-Lindau tumor suppressor protein: new insights into oxygen sensing and cancer. AB - The von Hippel-Lindau tumor suppressor protein (pVHL) is the substrate recognition module of an E3 ubiquitin ligase that targets the alpha subunits of hypoxia-inducible factor (HIF) for degradation in the presence of oxygen. Recognition of HIF by pVHL is linked to enzymatic hydroxylation of conserved prolyl residues in the HIF alpha subunits by members of the EGLN family. Dysregulation of HIF-target genes such as vascular endothelial growth factor and transforming growth factor alpha has been implicated in the pathogenesis of renal cell carcinomas and of hemangioblastomas, both of which frequently lack pVHL function. PMID- 12573437 TI - DNA repair defects in colon cancer. AB - Defects in DNA-repair pathways lead to an accumulation of mutations in genomic DNA that result from non-repair or mis-repair of modifications introduced into the DNA by endogenous or exogenous agents or by the malfunction of DNA metabolic pathways. Until recently, only two repair pathways, postreplicative mismatch repair and nucleotide excision repair, have been linked to cancer in mammals, but these have been joined in recent months also by the damage-reversal and base excision-repair processes, which have been shown to be inactivated, either through mutation or epigenetically, in human cancer. PMID- 12573438 TI - Telomeres and cancer: a tale with many endings. AB - Telomerase activity is necessary to maintain the integrity of telomeres, which in turn prevent chromosome ends from being processed and signaled as damaged DNA. That cancer cells rely on telomerase to maintain functional telomeres and to divide indefinitely has highlighted the potential for developing novel therapeutic approaches that target telomerase. PMID- 12573439 TI - Tumor suppression by Ink4a-Arf: progress and puzzles. AB - The two products of the Ink4a-Arf locus, p16(Ink4a) and p19(Arf) (p14(ARF) in humans), are potent tumor suppressors that regulate the activities of the retinoblastoma protein and the p53 transcription factor. These proteins form part of a signaling network that is disrupted in most, if not all, cancer cells. The Ink4a-Arf locus responds to stress signals, limiting cell proliferation and modulating oncogene-induced apoptosis. Recent evidence emerging from mouse tumor models distinguishes the activities of p16(Ink4a) and p19(Arf) in regulating tumor onset and identifies differences in their responsiveness to drugs. PMID- 12573440 TI - Mouse cancer models as a platform for performing preclinical therapeutic trials. AB - Engineering cancer-associated mutations into the mouse germline is an important tool for biological studies of growth control and tumorigenesis. Tractable models of many human cancers now exist in which the initiating genetic lesions have been elucidated and, in some instances, where the cooperating lesions are also known. The urgent need for more effective strategies for treating human cancer has stimulated interest in harnessing these models to test therapeutic agents. Although the ultimate value of genetically engineered mouse models for cancer drug discovery is unknown, several encouraging experiments provide proof of principle. PMID- 12573441 TI - Chemotherapy response and resistance. AB - Apoptosis and premature senescence, an acutely inducible terminal cell-cycle arrest, are known to be the ultimate cellular defense programs that counteract oncogenic transformation. Thus, activated oncogenes may sensitize cells to other stimuli that also recruit these programs. Recent evidence demonstrates that both apoptosis and premature senescence respond to drugs and can therefore contribute to the outcome of cancer therapy. However, manifest malignancies may have acquired mutations that compromise these programs at different levels, and hence may become chemoresistant to varying degrees as a result of defects in either or both programs. PMID- 12573442 TI - Classification of cancers by expression profiling. AB - Expression profiling using microarray technology has refined the classification of cancer. The greatest advances have been in lymphomas, leukaemias, and breast cancer where array information identified new diagnostic categories not achievable by standard microscopic or molecular means. These sub-categories often have distinct prognostic profiles. The same investigations have clarified the cellular lineage of cancer types, highlighted the importance of biochemical pathways in determining the expression 'phenotype', and identified potential new diagnostic markers and therapeutic targets. PMID- 12573443 TI - The OSBP-related proteins: a novel protein family involved in vesicle transport, cellular lipid metabolism, and cell signalling. AB - Proteins/genes showing high sequence homology to the mammalian oxysterol binding protein (OSBP) have been identified in a variety of eukaryotic organisms from yeast to man. The unifying feature of the gene products denoted as OSBP-related proteins (ORPs) is the presence of an OSBP-type ligand binding (LB) domain. The LB domains of OSBP and its closest homologue bind oxysterols, while data on certain other family members suggest interaction with phospholipids. Many ORPs also have a pleckstrin homology (PH) domain in the amino-terminal region. The PH domains of the family members studied in detail are known to interact with membrane phosphoinositides and play an important role in the intracellular targeting of the proteins. It is plausible that the ORPs constitute a regulatory apparatus that senses the status of specific lipid ligands in membranes, using the PH and/or LB domains, and mediates information to yet poorly known downstream machineries. Functional studies carried out on the ORP proteins in different organisms indicate roles of the gene family in diverse cellular processes including control of lipid metabolism, regulation of vesicle transport, and cell signalling events. PMID- 12573444 TI - Aging and acyl-CoA binding protein alter mitochondrial glycerol-3-phosphate acyltransferase activity. AB - It is well known that cellular function declines with age. Since phosphatidic acid (PtdOH) biosynthesis is central to the generation of membrane phospholipids, the hypothesis that aging decreases PtdOH biosynthesis was tested. Glycerol-3 phosphate acyltransferase (GPAT) and lysophosphatidic acid acyltransferase (LAT) activities were examined in isolated mitochondria and microsomes from young and old rat liver. The results show that mitochondrial GPAT preference for palmitoyl CoA over oleoyl-CoA was only observed if albumin or acyl-CoA binding protein (ACBP) were present in the assay in the young rats. Furthermore, mitochondrial GPAT activity was significantly reduced in the presence of albumin and ACBP in aged mitochondria using palmitoyl-CoA as the substrate. These data show, for the first time, that mitochondrial GPAT acyl-CoA preference is due to the presence of a protein that binds acyl-CoAs, not the enzyme itself, and that aging significantly reduces mitochondrial GPAT activity. PMID- 12573445 TI - Partial purification and characterization of phosphatidic acid-specific phospholipase A(1) in porcine platelet membranes. AB - We have shown previously that the phospholipase A (PLA) activity specific for phosphatidic acid (PA) in porcine platelet membranes is of the A(1) type (PA PLA(1)) [J. Biol. Chem. 259 (1984) 5083]. In the present study, the PA-PLA(1) was solubilized in Triton X-100 from membranes pre-treated with 1 M NaCl, and purified 280-fold from platelet homogenates by sequential chromatography on blue Toyopearl, red-Toyopearl, DEAE-Toyopearl, green-agarose, brown-agarose, polylysine-agarose, palmitoyl-CoA-agarose and blue-5PW columns. In the presence of 0.1% Triton X-100 in the assay mixture, the partially purified enzyme hydrolyzed the acyl group from the sn-1 position of PA independently of Ca(2+) and was highly specific for PA; phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositol (PI) were poor substrates. The enzyme exhibited lysophospholipase activity for l acyl-lysoPA at 7% of the activity for PA hydrolysis but no lipase activity was observed for triacylglycerol (TG) and diacylglycerol (DG). At 0.025% Triton X 100, the enzyme exhibited the highest activity, and PA was the best substrate, but PE was also hydrolyzed substantially. The partially purified PA-PLA(1) in porcine platelet membranes was shown to be different from previously purified and cloned phospholipases and lipases by comparing the sensitivities to a reducing agent, a serine-esterase inhibitor, a PLA(2) inhibitor, a Ca(2+)-independent phospholipase A(2) inhibitor, and a DG lipase inhibitor. PMID- 12573446 TI - Genomic structure and expression of a gene coding for a new fatty acid binding protein from Echinococcus granulosus. AB - This work describes a new gene coding for a fatty acid binding protein (FABP) in the parasite Echinococcus granulosus, named EgFABP2. The complete gene structure, including the promoter sequence, is reported. The genomic coding domain organisation of the previously reported E. granulosus FABP gene (EgFABP1) has been also determined. The corresponding polypeptide chains share 76% of identical residues and an overall 96% of similarity. The two EgFABPs present the highest amino acid homologies with the mammalian FABP subfamily containing heart-FABPs (H FABPs). The coding sequences of both genes are interrupted by a single intron located in the position of the third intron reported for vertebrate FABP genes. Both genes are expressed in the protoscolex stage of the parasite. The promoter region of EgFABP2 presents several consensus putative cis-acting elements found in other members of the family, suggesting interesting possible mechanisms involved in the host-parasite adaptation. PMID- 12573447 TI - Novel prostaglandin D(2)-derived activators of peroxisome proliferator-activated receptor-gamma are formed in macrophage cell cultures. AB - Incubation of RAW 264.7 murine macrophages with 9,15-dihydroxy-11-oxo-, (5Z,9alpha,13E,15(S))-Prosta-5,13-dien-1-oic acid [prostaglandin D(2) (PGD(2))] induced formation of considerable peroxisome proliferator-activated receptor gamma (PPARgamma) activity [Nature 391 (1998) 79]. Because PGD(2) itself is a poor PPARgamma ligand, we incubated RAW 264.7 macrophage cultures with prostaglandin D(2) for 24 h and studied the ability of the metabolites formed to activate PPARgamma. PGD(2) products were extracted and fractionated by reverse phase high-performance liquid chromatography. Chemical identification was achieved by UV spectroscopy, gas-liquid chromatography/mass spectrometry and chemical syntheses of reference compounds. PGD(2) was converted to eight products, six of which were identified. Ligand-induced interaction of PPARgamma with steroid receptor coactivator-1 was determined by glutathione-S-transferase pull-down assays and PPARgamma activation was investigated by transient transfection of RAW 264.7 macrophages. In addition to the previously known ligand 11-oxo-(5Z,9,12E,14Z)-Prosta-5,9,12,14-tetraen-1-oic acid (15-deoxy-delta(12,14) PGJ(2)), a novel PPARgamma ligand and activator viz. 9-hydroxy-11-oxo-, (5Z,9alpha,12E,14Z)-Prosta-5,12,14-trien-1-oic acid (15-deoxy-delta(12,14) PGD(2)) was identified. The biological significance of these results is currently under investigation. PMID- 12573448 TI - SR-BI does not require raft/caveola localisation for cholesteryl ester selective uptake in the human adrenal cell line NCI-H295R. AB - Class B type I scavenger receptor (SR-BI) mediates the selective uptake of high density lipoprotein (HDL)-derived cholesteryl esters (HDL-CE) in steroidogenic cells and hepatocytes. SR-BI is enriched in the caveolae of some cell types, genetically modified or not, and these domains have already been shown to constitute primary acceptors for HDL-CE. Nevertheless, the fate of caveola-free cell types has not yet been discussed.NCI-H295R, a human adrenal cell line, highly active in HDL-CE uptake via SR-BI, does not display any morphologically defined caveolae and expresses caveolin at a very low level. Using two different fractionation protocols, we have shown, in this cell type, that SR-BI is homogeneously distributed along the plasma membrane and consists principally of a non-raft membrane-associated pool. Raft destabilisation and caveolin-1 displacement from plasma membrane did not modify the SR-BI-mediated HDL-CE selective uptake. Moreover, the induction of SR-BI expression that is associated with increased CE selective uptake was not associated with any modification in caveolin-1 expression or any raft-targeting mechanism of SR-BI in NCI-H295R. In conclusion, we provide evidence that SR-BI does not require raft/caveola localisation to be implicated in CE selective uptake either in basal or in induced conditions. PMID- 12573449 TI - VLDL-induced triglyceride accumulation in human macrophages is mediated by modulation of LPL lipolytic activity in the absence of change in LPL mass. AB - Mixed dyslipidemia of phenotype IIB is characterized by elevated levels of very low density lipoprotein (VLDL)-1 and VLDL-2 subfractions and of low density lipoprotein (LDL), which are associated with premature formation of atherosclerotic plaques, characterized by the presence of lipid-rich macrophage foam cells. Lipoprotein lipase (LPL) is a key factor in mediating macrophage lipid accumulation and foam-cell formation from native VLDL particles. The action of macrophage-derived LPL in the induction of intracellular lipid accumulation from triglyceride-rich lipoprotein (TRL) subfractions (VLDL-1, VLDL-2) is, however, indeterminate, as is the potential role of VLDL-1 and VLDL-2 in modulating macrophage LPL expression. We evaluated the role of LPL in the interaction of type IIB VLDL-1 and VLDL-2 with human macrophages. Both VLDL-1 and VLDL-2 subfractions induced significant accumulation of triglyceride (9.8-fold, P<0.0001, and 4.8-fold, P<0.0001, respectively) and of free cholesterol content (1.4-fold, P<0.001, and 1.2-fold, P=0.02, respectively). Specific inhibition (90%) of the lipolytic activity of endogenous LPL by tetrahydrolipstatin (THL) in the presence of VLDL-1 or VLDL-2 resulted in marked reduction in cellular loading of both triglycerides (-89%, P=0.008, and -89%, P=0.015, respectively) and free cholesterol (-76%, P=0.02, and -55%, P=0.06 respectively). Furthermore, VLDL-1 and VLDL-2 induced marked increase in macrophage-derived LPL enzyme activity (+81%, P=0.002, and +45%, P=0.02), but did not modulate macrophage-derived LPL mRNA and protein expression; consequently, LPL specific activity was significantly increased from 1.6 mU/microg at baseline to 4.1 mU/microg (P=0.01) and 3.1 mU/microg (P=0.05), in the presence of VLDL-1 and VLDL-2, respectively. We conclude that type IIB VLDL-1 and VLDL-2 induce triglyceride accumulation in human monocyte-macrophages primarily via the lipolytic action of LPL, which may involve stabilization and activation of the macrophage-secreted enzyme, rather than via modulation of enzyme production. PMID- 12573451 TI - Natural mutations of apolipoprotein A-I impairing activation of lecithin:cholesterol acyltransferase. AB - Five natural mutations of apolipoprotein A-I (apoA-I), apoA-I(A95D), apoA I(Y100H), apoA-I(E110K), apoA-I(V156E) and apoA-I(H162Q), were studied for their ability to activate lecithin:cholesterol acyltransferase (LCAT). Mutants apoA I(E110K), apoA-I(V156E) and apoA-I(H162Q) had an impaired ability to activate LCAT. Combined with data on other apoA-I mutants this finding is consistent with the idea that the central region between amino acids 110 and 160 is likely to be the "active site" of apoA-I involved in the interaction with LCAT and that a specific sequence of apoA-I is required for activation of the enzyme. PMID- 12573450 TI - Nuclear phosphoinositide 3-kinase C2beta activation during G2/M phase of the cell cycle in HL-60 cells. AB - The activity of nuclear phosphoinositide 3-kinase C2beta (PI3K-C2beta) was investigated in HL-60 cells blocked by aphidicolin at G(1)/S boundary and allowed to progress synchronously through the cell cycle. The activity of immunoprecipitated PI3K-C2beta in the nuclei and nuclear envelopes showed peak activity at 8 h after release from the G(1)/S block, which correlates with G(2)/M phase of the cell cycle. In the nuclei and nuclear envelopes isolated from HL-60 cells at 8 h after release from G(1)/S block, a significant increase in the level of incorporation of radiolabeled phosphate into phosphatidylinositol 3-phosphate (PtdIns(3)P) was observed with no change in the level of radiolabeled PtdIns(4)P, PtdIns(4,5)P(2) and PtdIns(3,4,5)P(3). On Western blots, PI3K-C2beta revealed a single immunoreactive band of 180 kDa, whereas in the nuclei and nuclear envelopes isolated at 8 h after release, the gel shift of 18 kDa was observed. When nuclear envelopes were treated for 20 min with mu-calpain in vitro, the similar gel shift and increase in PI3K-C2beta activity was observed which was completely inhibited by pretreatment with calpain inhibitor calpeptin. The presence of PI3K inhibitor LY 294002 completely abolished the calpain-mediated increase in the activity of PI3K-C2beta but did not prevent the gel shift. When HL-60 cells were released from G(1)/S block in the presence of either calpeptin or LY 294002, the activation of nuclear PI3K-C2beta was completely inhibited. These results demonstrate the calpain-mediated activation of the nuclear PI3K C2beta during G(2)/M phase of the cell cycle in HL-60 cells. PMID- 12573452 TI - Inhibition by eicosapentaenoic acid of IL-1beta-induced PGHS-2 expression in human microvascular endothelial cells: involvement of lipoxygenase-derived metabolites and p38 MAPK pathway. AB - Prostaglandin H synthase 2 (PGHS-2), a highly inducible isoenzyme, is responsible for overproduction of the prostaglandins (PGs) in inflammatory sites. We established that among fish oil polyunsaturated fatty acids (PUFAs), eicosapentaenoic acid (EPA), but not docosahexaenoic acid (DHA), greatly decreased interleukin-1beta (IL-1beta)-induced PGHS-2 expression in human pulmonary microvascular endothelial cells (HPMECs). Lipoxygenase products 12 (S) hydroperoxyeicosapentaenoic acid ((S)-HpEPE), 15 (S)-HpEPE and leukotriene (LT) D5 reproduced similar inhibitory effect, suggesting that they may be the intermediate metabolites responsible for PGHS-2 down-regulation by EPA. Accordingly, the EPA effect is prevented by nordihydroguaiaretic acid (NDGA) and by REV 5901, nonspecific and specific 5-lipoxygenase inhibitors, respectively. Besides, inhibition of cyclooxygenase activity by ibuprofen, indomethacin or aspirin was not able to prevent this effect. Moreover, cyclooxygenase metabolites of EPA (PGs D3, E3 and I3) markedly potentiate IL-1beta-induced PGHS-2 expression, probably by increasing intracellular cAMP levels. Peroxisome proliferator-activated receptors (PPARs) are known to be activated by fatty acids (FAs) such as EPA. We found here that HPMECs express only weak amounts of PPARalpha and PPARgamma whose activation by synthetic agonists, Wy-14,643 and ciglitazone, does not cause any inhibition of IL-1beta-induced PGHS-2 expression. This finding ruled out the involvement of PPARs in the EPA inhibitory effect. In addition, we established that EPA, which failed to inhibit nuclear factor-kappaB (NF-kappaB) activation, suppressed p38 mitogen-activated protein kinase (MAPK) phosphorylation in stimulated HPMECs. Our data demonstrate that EPA, unlike DHA, down-regulates PGHS-2 expression in HPMECs probably through its 5-lipoxygenase dependent metabolites and advocates a beneficial role for this FA in limiting inflammatory response. PMID- 12573453 TI - Induction of 5-lipoxygenase activation in polymorphonuclear leukocytes by 1 oleoyl-2-acetylglycerol. AB - 1,2-Diacylglycerols (DAGs) can prime polymorphonuclear leukocytes (PMNL) for enhanced release of arachidonic acid (AA) and generation of 5-lipoxygenase (5-LO) products upon subsequent agonist stimulation. Here, we demonstrate that in isolated human PMNL, 1-oleoyl-2-acetylglycerol (OAG) functions as a direct agonist stimulating 5-LO product formation (up to 42-fold). OAG caused no release of endogenous AA, but in the presence of exogenous AA, the magnitude of 5-LO product synthesis induced by OAG was comparable to that obtained with the Ca(2+) ionophore A23187. Interestingly, OAG-induced 5-LO product synthesis was not connected with increased 5-LO nuclear membrane association. Examination of diverse glycerides revealed that the sn-2-acetyl-group is important, thus, also 1 O-hexadecyl-2-acetylglycerol (EAG) stimulated 5-LO product formation (up to 8 fold). Treatment of PMNL with OAG did not alter the mobilization of Ca(2+) but removal of intracellular Ca(2+) abolished the upregulatory OAG effects. Notably, the PKC activator phorbol-myristate-acetate hardly increased 5-LO product synthesis and PKC inhibitors failed to suppress the effects of OAG. Although OAG rapidly activated p38 MAPK and p42/44(MAPK), which can stimulate 5-LO for product synthesis, specific inhibitors of these kinases could not prevent 5-LO activation by OAG. Together, OAG acts as a direct agonist for 5-LO product synthesis in PMNL stimulating 5-LO by novel undefined mechanisms. PMID- 12573454 TI - Characterization and biological activity of gangliosides in buffalo milk. AB - Gangliosides (GS) were evaluated in Swiss cow's milk (SCM), Italian buffalo milk (IBM) and its serum, Pakistan buffalo colostrum (PBC), Pakistan buffalo mature milk (PBM), and Pakistan buffalo milk from rice-growing areas (PBR). Dairy GS were obtained from the Folch's upper (hydrophilic) and lower (lipophilic) extraction phases, respectively, and determined as lipid-bound sialic acid (LBSA) by colorimetry. Molar ratios of LBSA in the hydro- and lipophilic GS fractions were 52:48 to 79:21. Mature buffalo milk types had 40-100% more LBSA in the lipophilic GS fraction compared to SCM. Liquid PBC was higher in LBSA (24 nmol/g) compared to mature milk types (8-11 nmol/g). Thin-layer chromatography (TLC) and scanning densitometry showed distinct profiles of hydrophilic and lipophilic GS fractions. Lipophilic GS (but importantly not hydrophilic GS) from IBM and its serum decreased prostaglandin series 2 production by 75-80% in cultured human colonic epithelial cells exposed to tumor necrosis factor alpha (TNFalpha). Hydrophilic GD(3) and lipophilic GM(3) selectively bound rotavirus particles prepared from a rhesus strain and its mutant. A GS fraction in IBM showed a GM(1) specific binding to cholera toxin subunit B (CTB). IBM serum (IBMS) was a rich source of LBSA (420 nmol/g proteins). In summary, improved methodology led to increased LBSA recovery and isolation of additional and bioactive milk GS. Human and Italian buffalo milk had similar CTB binding, and both had increased polysialo-GS compared to cows milk. The toxin binding properties of buffalo milk GS, and the anti-inflammatory activity of the lipophilized GS fraction could be important for developing innovative food applications, as well as the subject of future research. PMID- 12573455 TI - The GABA(A) receptor complex in the chicken brain: immunocytochemical distribution of alpha 1- and gamma 2-subunits and autoradiographic distribution of BZ1 and BZ2 binding sites. AB - Two antibodies, raised against the rat GABA(A) receptor alpha1- and gamma2 subunits, were used for an immunocytochemical study of the distribution of these proteins in the chicken brain. The immunoreactive bands obtained by Western blotting and the similar labelling distribution found in the rat and chicken brain support the suitability of these antibodies for the labelling of GABA(A) receptors in birds. We found abundant alpha1 and gamma2 immunoreactivity throughout the chicken brain, mainly in the paleostriata and lobus paraolfactorius, dorsal thalamus and some nuclei of the brainstem. The alpha1 subunit was more abundant in the telencephalon, thalamus and cerebellum, while the presence of the gamma2-subunit was stronger in the optic tectum and brainstem. We also report the autoradiographic distribution of the BZ1 and BZ2 benzodiazepine receptor subtypes in the chicken brain using [3H]flunitrazepam. Benzodiazepine binding was unevenly distributed throughout the chicken brain, and the anatomical distribution of the BZ1 and BZ2 subtypes was similar to that described in mammals. The highest binding values were found in the olfactory bulb, paleostriatum primitivum, optic tectum, nucleus mesencephalicus lateralis pars dorsalis and nucleus isthmi pars parvocellularis, the BZ2 subtype being predominant in the paleostriatum primitivum and optic tectum. A general agreement in the distribution of BZ1 and alpha1 immunoreactivity was observed in structures such as the olfactory bulb, paleostriata, lobus parolfactorius and dorsal thalamus, although some discrepancies were observed in areas such as the optic tectum or nucleus isthmi pars parvocellularis, with high BZ1 binding and low or no alpha1 immunolabelling. PMID- 12573456 TI - Catecholaminergic and dopamine-containing neurons in the spinal cord of pigeons: an immunohistochemical study. AB - Within the different species belonging to the vertebrate radiation, catecholaminergic elements of the spinal cord present a partly conservative, partly variable pattern. Unfortunately, the overall picture is far from clear since the situation for birds is largely obscure. Therefore, we examined the distribution of dopamine (DA)- and tyrosine hydroxylase (TH)-positive cells and fibers in the spinal cord of the adult pigeon by immunohistochemistry. TH immunoreactive cells were located within two restricted areas. One group of cells with multipolar shape was located in laminae VI and VII, close to the white-gray border. These cells were more frequently found at rostral and caudal levels while being scarce at cervical-thoracic levels. The second group of cells was located in lamina VIII surrounding the central canal. These cells were bipolar in shape and were found ventrally and laterally to the central canal, with most of them contacting the lumen of the canal through a separate process. The TH immunoreactive fibers were distributed in both the gray and the white matter. In the gray matter, they were mainly distributed around the central canal (lamina VIII), in the ventral horn close to the border of laminae VII-IX and in the lateral part of the dorsal horn in laminae II-VI. In the white matter the fibers were present in the lateral columns running longitudinal to the main axis. DA immunoreactive cells were also located within two restricted areas, closely matching the distribution of TH-immunopositive ones. Additionally, the DA immunoreactive cells had the same shape as the TH-immunoreactive cells, as bipolar neurons contacted the central canal and multipolar ones were located in the laminae VI and VII. Also the distribution of DA- and TH-immunoreactive fibers roughly matched. Both, DA-immunoreactive cells and fibers were scarcer than TH immunoreactive ones. This finding suggests that the catecholaminergic system in the spinal cord consists of DA-immunoreactive cells as well as other catecholaminergic cells. PMID- 12573457 TI - Evidence for the presence of both peroxisome proliferator-activated receptors alpha and beta in the rat spinal cord. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors belonging to the nuclear receptor superfamily. Different subtypes of PPARs (alpha, beta, and gamma) have been described. Their distinct physiological functions depend on their differential ligand activation profiles but also on their specific tissue expression. Previous studies have described their presence in the central nervous system. However, their expression in the adult rat spinal cord in normal physiological conditions has never been investigated. We demonstrated by using reverse-transcription-polymerase chain reaction, and Western blotting, the mRNA and protein expression of PPARalpha and PPARbeta, but not PPARgamma in cervical, thoracic, and lumbar segments of the spinal cord. Using immunohistochemistry, we also showed for the first time the specific cellular distribution of these transcription factors in the different segments of the spinal cord. In the gray matter, the distribution of PPARalpha was homogenous whereas PPARbeta was specifically localized in motoneurons and in medial part of laminae IV, V, VI, VII, VIII, and X. These latter areas are known as nociceptive afferent pathways to supra-spinal structures such as the medulla reticular nucleus and the thalamus. In the white matter, PPARalpha was localized exclusively in astrocytes while PPARbeta was present in oligodendrocytes. The possible functions of PPARalpha and PPARbeta expressed in both white and gray matters of the spinal cord will be discussed but need further studies. PMID- 12573458 TI - Anatomical relationships between aromatase-immunoreactive neurons and nitric oxide synthase as evidenced by NOS immunohistochemistry or NADPH diaphorase histochemistry in the quail forebrain. AB - In Japanese quail (Coturnix japonica), previous studies indicated that the distribution of reduced nicotinamide dinucleotide phosphate (NADPH) diaphorase overlaps with steroid-sensitive areas that contain dense populations of aromatase immunoreactive (ARO-ir) cells. We investigated here the anatomical relationships between aromatase (ARO) and nitric oxide synthase (NOS)-containing cells that were visualized both by NOS-immunohistochemistry and NADPH-histochemistry. The distribution of ARO-ir and of NADPH-positive cells in the forebrain observed here matched exactly the distribution previously reported. The distribution of NOS immunoreactive material in the vicinity of ARO-ir cell groups appeared similar to the distribution of NADPH-positive structures previously identified by histochemistry. The number of NOS-immunoreactive cells was similar to the number of NADPH-positive cells and they were found in the same brain regions. In contrast, few NOS-immunoreactive fibers were observed whereas numerous NADPH positive fibers and punctuate structures were present in many areas. Major groups of NOS-immunoreactive/NADPH-positive neurons were adjacent to the main ARO-ir cell groups, such as the medial preoptic nucleus, the bed nucleus of the stria terminalis and the nucleus ventromedialis hypothalamic. However, examination of adjacent sections indicated that there is very little overlap between the NOS immunoreactive and ARO-ir cell populations. This notion got further support by double-labeled sections where no double-labeled cells could be identified. In sections stained simultaneously by histochemistry for NADPH and immunohistochemistry for ARO, many NADPH-positive fibers and punctate structures were closely associated with ARO-ir perikarya. Taken together, the present data indicate that NOS is not or very rarely colocalized with ARO but that NOS inputs are closely associated with ARO-ir cells. Based on previous work in a variety of model systems, it can be hypothesized that these inputs modulate the expression or activity of ARO in the quail brain. PMID- 12573459 TI - Comparative analysis of neuropeptide FF-like immunoreactivity in the brain of anuran (Rana perezi, Xenopus laevis) and urodele (Pleurodeles waltl) amphibians. AB - The neuropeptide FF (NPFF) is a member of the RFamide related peptides (FaRPs) that share the dipeptide Arg-Phe-NH2 at their C-terminal. It was originally isolated from bovine brain and its wide distribution has been demonstrated in the brain of several mammalian species. By means of an NPFF antiserum we have investigated the distribution pattern of NPFF-like immunoreactive cells and fibers in the brain of anuran and urodele amphibians. In both amphibian orders, the most conspicuous labeled cell population was found in the preoptic area and hypothalamus, primarily in the suprachiasmatic region. Numerous fibers reached the median eminence and the intermediate lobe of the hypophysis. Only in the anuran brain cells were observed in the pallium and septum. In the urodele, cells and fibers of the terminal nerve were distinctly labeled. Cell bodies were widely distributed in the reticular formation of anurans and, in both orders, a large cell population was found in the nucleus of the solitary tract and the spinal cord. Comparable fiber distribution between both orders exists in which the basal telencephalon (mainly the amygdaloid complex), the hypothalamus and the spinal cord are the regions most richly innervated. The distribution pattern of NPFF like immunorective elements in the brain of amphibians, which only partly overlaps with those of other FaRPs, supports the notion that a NPFF-like peptide exists in amphibians. On the basis of its localization, this peptide may act as a hypophysiotropic neurohormone and be involved in background adaptation. Its wide distribution in similar zones of the brain in amphibians and mammals suggests that functional roles of this peptide have been conserved in vertebrate evolution. PMID- 12573460 TI - Localization of cyclic nucleotide phosphodiesterase 2 in the brain-derived Triton insoluble low-density fraction (raft). AB - The cyclic nucleotides perform a variety of roles in the formation and remodeling of the neuronal interaction. The membrane microdomain called "raft" has been paid much attention, for this domain contains many signal-transducing molecules including trimeric G proteins and cytoskeletal proteins. The raft domain is recovered in a low-density fraction after the treatment of the membrane with a non-ionic detergent such as Triton X-100. The enrichment of cholesterol and sphingolipids is ascribed to be responsible for the detergent insolubility. In this study we focused on the cyclic nucleotide signaling process in rafts prepared from the cerebral cortex of 10-day-old rat and the synaptic plasma membrane fraction and found the presence of a high cAMP and cGMP phosphodiesterase (PDE) activity. The activity was effectively inhibited with erythro-9-(2-hydroxy-3-nonyl)adenine, a PDE2-specific inhibitor but not with other inhibitors such as vinpocetine, quazione, or zaprinast. Further western blotting analysis confirmed the localization of PDE2 in the raft fraction. The presence of adenylyl cyclase V/VI and PKA in the raft fraction was also shown with Western blotting. These results suggest the participation of the raft in the cyclic nucleotide signaling cascade in neurons. PMID- 12573461 TI - Cortical spreading depression affects Fos expression in the hypothalamic paraventricular nucleus and the cerebral cortex of both hemispheres. AB - The present experiments were performed to clarify the brain sites whose activity is affected exclusively by cortical spreading depression (CoSD). For this purpose, Fos protein, a product of an immediate early gene, was used as a marker of neuronal activation. Because Fos can be induced by many manipulations such as stress stimuli, we verified CoSD-induced Fos expression by excluding the influence of other factors such as anaesthesia and surgical manipulation. CoSD was induced by applying a KCl solution directly to the dura mater over the cerebral cortex, and Fos expression in the brain was assessed by immunohistochemistry using antibodies against Fos protein. We found that during CoSD, Fos expression was increased specifically in the magnocellular region of the hypothalamic paraventricular nucleus (PVN), as well as in the ipsilateral cortex, whereas reduced Fos expression was observed in both the parvocellular region of the PVN and the whole cortex contralateral to the CoSD site. Consistent with the reduced Fos expression, approximately 40% of neurons in the contralateral cortex revealed a suppression of electrical activity during CoSD. These results suggest that in addition to the ipsilateral cortex, CoSD affects Fos expression exclusively in the PVN and the contralateral cortex. PMID- 12573462 TI - Blockade of peroxynitrite-mediated astrocyte death by manganese(III)-cyclam. AB - Under glucose-deprived conditions, astrocytes rapidly underwent death due to their increased susceptibility to endogenously produced peroxynitrite (Gila 31, 155-164; J. Neuroimmunol. 112, 55-62; J. Neurochem. 74, 1989-1998). In the present study, the cell membrane-permeable synthetic superoxide dismutase (SOD) mimetic cyclam manganese(III) 1,4,8,11-tetraazacyclodecane (Mn(III)-cyclam) completely inhibited the death of glucose-deprived immunostimulated astrocytes. However, the structurally related compounds Ni(II)-cyclam, Co(II)-cyclam, and H(2)-cyclam, which lacks metals, had no or a little cytoprotective effect. Of the cyclams used in this study, only Mn(III)-cyclam completely scavenged the peroxynitrite produced in glucose-deprived immunostimulated astrocytes and significantly blocked the depolarization of mitochondrial transmembrane potential in those cells. The present results suggest that cell membrane-permeable synthetic SOD mimetics such as Mn(III)-cyclam may be potential therapeutic agents for various diseases associated with the endogenous production of peroxynitrite. PMID- 12573463 TI - Intrinsic differences in sensitivity to 5-HT between high- and low-output terminals innervating the same target. AB - The differential action of neuromodulators on synapses of various efficacy provides additional fine tuning of synaptic regulation beyond frequency induced plasticity. We used the well-characterized high- and low-output motor nerve terminals, of the tonic and phasic neuromuscular junctions (NMJs) in the walking leg extensor muscle of the crayfish, to investigate differential actions of serotonin (5-HT) since both terminals innervate the same target. The excitatory postsynaptic potentials of the tonic NMJ are enhanced to a greater extent than for the phasic NMJs during exposure to 5-HT (100 nM). Macropatch current recordings at identified sites along the motor nerve terminals and quantal analysis indicate that mean quantal content is substantially increased by 5-HT. The overall probability of vesicular release increases to a greater extent at tonic terminals than at phasic terminals when exposed to 100 nM 5-HT. Measures in the area (i.e. charge) of spontaneous quantal currents indicate no difference in postsynaptic receptivity to the glutamatergic synaptic transmission upon exposure to 5-HT. The results provide new details concerning differential modulation of low- and high-output synapses present on the same target tissue. PMID- 12573464 TI - Exogenous expression of hepatocyte growth factor (HGF) in rat striatum by naked plasmid DNA. AB - Hepatocyte growth factor (HGF) is a heterodimeric protein and shows mitogenic and morphogenic activities toward a variety of epithelial cells. There has been no immunohistochemical evidences for naked DNA mediated transgene expression of HGF into central nervous system. We initially demonstrated a naked plasmid mediated expression of HGF into rat striatum. The immunofluorescence staining revealed that exogenous protein of human HGF was expressed at 7 days after plasmid injection (300 microg). Exogenous HGF was mainly expressed in reactive astrocytes according to dual-labeling staining of HGF and glial fibrillary acidic protein or S100. It is also demonstrated that c-met, specific receptor of HGF, was expressed in the injection site. Intensive expression of c-met was found in the site to which HGF encoded plasmid was injected. These evidences for the exogenous expression of HGF and its receptor c-met may implicate an application of naked plasmid mediated HGF for neuronal disease as well as the other neurotrophic factors. PMID- 12573465 TI - Changes in neurocan expression in the distal spinal cord stump following complete cord transection: a comparison between infant and adult rats. AB - The distal transected cords of infant rats are more permissive for axon extension than those of adults. To elucidate the biomolecular basis for this phenomenon, we examined the expression pattern of neurocan using semi-quantitative reverse transcription polymerase chain reaction and immunostaining in the distal cord of both adult and infant rats after transection. Neurocan is a chondroitin sulfate proteoglycan with well-documented axon growth-inhibitory properties in the central nervous system. Neurocan mRNA was up-regulated in the distal cord of adult rats shortly after transection, followed by a longer wide distribution of neurocan immunoreactivity (IR) in both neurons and astrocytes; by contrast, upregulation of neurocan mRNA was not seen in infant rats, although transient expression of neurocan IR was seen in neurons. Combined with the different regenerative capacity of infant and adult rats, the present results suggest that neurocan inhibits spinal cord regeneration. PMID- 12573466 TI - Neuronal gap junctions in the rat main olfactory bulb, with special reference to intraglomerular gap junctions. AB - The structural features of neuronal gap junction-forming processes in the rat olfactory bulb were analyzed electron microscopically. Gap junctions were present in glomeruli and extraglomerular regions. In extraglomerular regions, mitral/tufted cell somata, dendrites and axon hillock-initial segments made gap junctions and mixed synapses with interneuronal processes, some of which were confirmed to be GABA positive. In glomeruli gap junctions were encountered mainly between mitral/tufted cell dendrites and diverse types of processes; a small population of them were conclusively identified as periglomerular cell dendrites. Gap junction-forming processes frequently received synapses from olfactory nerve terminals, suggesting that they could be type 1 periglomerular cells. However, the majority were GABA negative or only faintly positive and none were tyrosine hydroxylase positive, indicating that they were different from previously reported type 1 periglomerular cells. Furthermore serial sectioning analyses revealed that the majority of those processes forming gap junctions with mitral/tufted dendrites were smooth cylindrical and had few presynaptic sites, indicating that they were different from previously described periglomerular cells. These findings revealed that mitral/tufted cells make gap junctions with diverse types of neurons; and some of these gap junction-forming processes originated from some types of periglomerular cells but others from hitherto uncharacterized neuron type(s). PMID- 12573467 TI - Effect of target saliency on human smooth pursuit initiation: interocular transfer. AB - Our previous study showed that the saliency of a target increases the gain of smooth pursuit initiation. In this study, we examined the interocular transfer of this effect in five humans. A square red frame surrounding the target was used as a cue to indicate the initial target position. In the cue condition, the responses were similar, irrespective of the eye to which the cue was presented, and were significantly larger than in the no-cue condition. The result suggests that central pathways that receive input from both eyes mediate the effect of saliency on smooth pursuit initiation. PMID- 12573468 TI - Receptor-type protein tyrosine phosphatase zeta as a component of the signaling receptor complex for midkine-dependent survival of embryonic neurons. AB - Midkine (MK), a heparin-binding growth factor, suppresses apoptosis of embryonic neurons in culture, induced by serum deprivation. Receptor-type protein tyrosine phosphatase zeta (PTP zeta) is a chondroitin sulfate proteoglycan with a transmembrane domain and intracellular tyrosine phosphatase domains. The activity of MK was abolished by digestion with chondroitinase ABC, or addition of the antibody to PTP zeta, while digestion with heparitinase showed no significant effect. These results suggested that the survival-promoting signal of MK was received by a receptor complex containing PTP zeta. Low density lipoprotein receptor-related protein (LRP) has been identified as another component of the signaling receptor. Ectodomains of two related proteins expressed on neurons, namely LRP6 and apoE receptor 2, were FLAG-tagged and examined for MK binding, using MK-agarose column. Both the ectodomains were found to exhibit calcium dependent binding to MK. These proteins may participate in MK signaling in certain cases. The survival-promoting activity of MK was abolished by PP1, an inhibitor of src protein kinase, pertussis toxin, an inhibitor of G protein linked signaling and sodium orthovanadate, an inhibitor of PTPs. PMID- 12573469 TI - Toosendanin induces outgrowth of neuronal processes and apoptosis in PC12 cells. AB - In the present study, the effects of toosendanin on cell differentiation and apoptosis were investigated in PC12 cells. The results showed that after 24-48 h of culture in a medium containing toosendanin (approximately 1-10x10(-7) M), cell differentiation and outgrowth of neuronal processes were promoted. Combined treatment with toosendanin and a calcium channel blocker, nifedipine or omega conotoxin GVIA, resulted in a significant inhibition of the toosendanin-induced effects. Pretreatment of PC12 cells with BAPTA-AM also inhibited the toosendanin induced effects; however, these effects were not inhibited by pertussis toxin and H-7 in the medium. Toosendanin also induced cell apoptosis. Based on the DNA content determined by flow cytometric analysis, the number of apoptotic cells significantly increased when the incubation time in the toosendanin-containing medium was lasted up to 72 h. Toosendanin at a higher concentration (> or =1 x 10(-6) M) caused cell death while it had no effect on cell division at concentrations lower than 1 x 10(-7) M. PMID- 12573470 TI - Regulated transgene delivery by ganciclovir in the brain without physiological alterations by a live attenuated herpes simplex virus vector. AB - The distribution of a live attenuated herpes simplex virus (betaH1)-mediated gene delivery into the central nervous system (CNS) was regulated by growth inhibition with ganciclovir (GCV) and the effect of this transgene expression system on the physiologic response was characterized by the acoustic startle response and its prepulse inhibition. We inoculated betaH1 expressing beta-galactosidase (beta gal) driven by the latency associated transcripts promoter into the right caudate putamen of rats. Histochemical analysis demonstrated that the inoculation of betaH1 in the right caudate putamen resulted in a high level of beta-gal expression in the neurons of the area projecting to the inoculation site. On 14 days after inoculation without GCV-treatment, beta-gal activity localized in the anterior olfactory nucleus, frontal, insular, orbital, parietal, perirhinal, piriform cortices and the temporal region including the amygdala. In contrast, the distribution of beta-gal activity was regulated by the interval between virus inoculation and GCV-treatment and maintained after its cessation without significant alteration. The whole process of transgene expression did not influence the emotional behavior, indicating that this vector system is a suitable model for analyzing the transgene function or applying the gene therapy for the CNS diseases. PMID- 12573471 TI - Mapping of presynaptic nicotinic acetylcholine receptors using fluorescence imaging of neuritic calcium. AB - Neuronal nicotinic receptors (nAChRs) appear to function at both pre- and postsynaptic sites, to modulate the release of neurotransmitter, and to mediate synaptic transmission, respectively. Localization of functional nAChRs at presynaptic structures has only been possible under the best of circumstances where the presynaptic structure is very large allowing direct nAChR channel recording. We report here a novel and simple method that allows the visualization of stimulus-evoked changes in Fura-2 fluorescence in the presynaptic structures of essentially any neuron type in vitro. Following 'loading' of all neurons by incubation with the calcium-sensitive dye, Fura-2-AM, we selectively reduced the fluorescent signal in the postsynaptic neuron by injecting the Fura-2 quenching agent, Mn(2+), into the postsynaptic neuron. After quenching, nicotine treatment elicits calcium transients that can be observed in spatially distinct regions of neurite bundles contacting the Mn(2+)-infused neuron. Thus, the approach described allows one to readily map the distribution of activated nAChRs on presynaptic inputs in vitro. PMID- 12573472 TI - Closed-loop multivariable system identification for the characterization of the dynamic arm compliance using continuous force disturbances: a model study. AB - This study presents a new multivariable closed-loop identification technique for estimating the dynamic compliance of the multijoint human arm during posture maintenance. The method is designed for the application of continuous force disturbances that facilitate interaction of the limb with the environment. The dynamic compliance of the arm arises from different physiological mechanisms and is important for maintaining stable postures and to suppress disturbances. Estimates can be useful to analyze the ability of the nervous system to adapt the arm compliance to different types of disturbances and environments. The technique is linear and requires no a priori knowledge of the system. Linear system behavior is justified for posture tasks where the hand position deviates slightly from a reference position. Interaction results in a closed-loop configuration of arm and environment. The problem with previous methods is the restriction to open loop systems. With the current technique, the dynamic arm compliance is separately estimated from the closed-loop. The accuracy of the identification technique is tested by simulations for different values of the dynamic compliance of the arm and environment and for different methodological parameters. It is concluded that the identification technique is accurate, even for short observation periods and severe noise. PMID- 12573473 TI - Detection of single unit activity from the rat vagus using cluster analysis of principal components. AB - In vivo recordings from subdiaphragmatic vagal afferent nerves generally lack the resolution to distinguish single unit activity. Several methods for data acquisition and analysis were combined to produce a high degree of reliability in recording electrophysiological signals from gastrointestinal and hepatic afferent fibers in the rat. Recordings with low noise were achieved by paralysis of the respiratory muscles and by pinning the nerve to a recording platform. Single unit activity was isolated using principal component (PC) analysis and cluster cutting of data in multi-dimensional space (1-3 PCs). Cluster assignments were determined by a semi-automated approach using the k-means algorithm. The accuracy of single unit classification was assessed by checking inter-spike intervals (ISIs) to determine the length of the refractory period, and by cross-correlation analysis to assess whether single units were mistakenly split into more than one cluster. These analyses produced up to four isolated single units from each nerve filament (a bundle of nerve fibers), and typically it was possible to further increase yield by recording from several nerve filaments simultaneously using an array of electrodes. PMID- 12573474 TI - Development of a novel fluorescent microsphere technique to combine serial cerebral blood flow measurements with histology in the rat. AB - The aim of the study was to evaluate the microsphere technique for the quantitative assessment of regional cerebral blood flow (rCBF) at different time points in the same animal. Yellow-green and red fluorescent microspheres with a diameter of 15 microm were injected into the rat at two different time points via a cannula inserted into the left ventricle of the heart. The reproducibility of the rCBF measurements in normocapnic conditions (n=7) and the responsiveness of the flow to hypercapnia induced by 7% CO(2) (n=7) was examined. The fluorescent spheres were counted on 100 microm vibratome sections of perfusion-fixed brains and rCBF was calculated. The median total CBF in normocapnic rats was 224 ml/min/100 g for the first microsphere injection and 216 ml/min/100 g for the second one. In the hypercapnic group CBF amounted to 400 ml/min/100 g and after 30 min of normocapnia decreased to 178 ml/min/100 g. No differences between the left and right hemisphere were found and there was no indication that the first injection might have influenced the second one. The described approach allows combining the assessment of rCBF at different time points in physiological or pathological conditions with histological evaluation of related morphological alterations in the same brain region of the same animal. PMID- 12573475 TI - Extending the resolution of light microscopy and electron microscopy digitized images with reference to cellular changes after in vivo low oxygen exposure. AB - When processing frame-grabbed images from light microscopy (LM) and electron microscopy (EM), even a state-of-the-art digital camera is the weakest link between the microscope and the image processor. Details, which can be seen directly in the ocular at LM and in a negative recorded at EM, will not necessarily be represented in the frame-grabbed images. Because of this, there is a tendency to prefer a higher magnification at the expense of overview, i.e. only smaller areas are described. We find that the inadequacy of the camera can be overcome by taking multiple images of the same object, and align, expand, and add them into a more highly resolved image. At the LM level, the method has proved useful for describing the relation of the zinc pattern versus local in vivo oxygen measurements. At the EM level, we show that it is possible to achieve information about the spatial conditions in a given area, and the method may have applications on, e.g., visualization of ultra-small antibody-bound gold particles. The method can be performed on color and black and white images at any magnification and it has been tested in Adobe Photoshop (4.0 and higher) in WINDOWS 95, 98, and 2000. PMID- 12573476 TI - An injectable nerve regeneration chamber for studies of unstable soluble growth factors. AB - Modern surgical techniques cannot guarantee functional recovery following peripheral nerve injuries. Research into factors that may influence nerve regeneration has therefore assumed a prominent potential therapeutic role. We report here on the development of an approach to allow for direct manipulation of the microenvironment of regenerating peripheral nerve axons. We show that solutions can be delivered directly to this local milieu in vivo and that such a delivery can be performed multiple times over an extended period, potentially facilitating studies of multiple molecular players that act locally. We also demonstrate that the bundle of regenerated axons are amenable to morphological analysis by 21 days and that the injection system remains patent for at least 21 days. PMID- 12573477 TI - A method for quantifying reflex responses from intra-muscular and surface electromyogram. AB - Measuring human reflex responses from electromyogram (EMG) traces in an accurate, repeatable and reliable way with a high degree of specificity has traditionally been a difficult task. This paper describes a new method that can be used to quantify reflex responses from both surface and intra-muscular EMG. This technique extends the classical cumulative sum (CUSUM) calculations by defining precise points for the calculation of latencies, durations and strengths to facilitate automatic reflex detection and permit the strength of a reflex to be defined in absolute units. The effect of varying the pre-stimulus time, the number of trials averaged and the amount of filtering used on the identification and classification of reflex parameters are also investigated. Furthermore, the effect of noise on these values, and how to remove it, is discussed. The new method, which is an expansion of the CUSUM analysis, is compared and contrasted with the more common threshold-crossing method in two different muscles: masseter and first dorsal interosseous (FDI), in experiments utilizing both mechanical and electrical stimulation. There are a number of advantages to using the new method; not only does the modified CUSUM method detect reflexes earlier than threshold crossing methods but also the strength and duration are less susceptible to averaging and filtering parameters while giving a better indication of the reflex size. The data suggests that a pre-stimulus analysis period of at least 100 ms be used to correctly identify the variability inherent in EMG traces. It is also concluded that for subtle reflexes, 50 stimuli should be the minimum number used when spike trigger averaging is employed as lower numbers are associated with much greater pre-stimulus variability. Zero-phase filtering the rectified averaged EMG traces is recommended as this makes it easier to identify significant changes in the electrical activity of the muscle in question. In addition, noise estimation and removal from averaged rectified EMG recordings yields results that are a more accurate representation of the synaptic activity of the motor units in question. PMID- 12573478 TI - A novel technique to isolate adult Schwann cells for an artificial nerve conduit. AB - The use of an artificial nerve conduit containing viable Schwann cells (SCs) is one of the most promising approaches to repair nerve injuries. Obtaining a large number of viable SCs in a short period is demanded for the clinical use of this technique. However, the previous methods using mitogens are not clinically acceptable, and other methods that do not require mitogens, failed to isolate adult SCs effectively or required a long period of time. In this study, we have developed a novel technique to isolate SCs from adult rat peripheral nerves for an artificial nerve conduit without mitogens, which has produced a total number of 1.21 x 10(5) cells per mg, with an average purity of 93.0+/-0.58% at 21 days in vitro. The Bottenstein-Sato (BS) medium used in this study, had originally been developed for oligodendrocyte culture, but here it is shown to have an effect on SC proliferation and survival. By changing fetal bovine serum (FBS) concentrations from 0 to 10% serially, SCs could be isolated maximally from the predegenerated nerves while suppressing fibroblast overgrowth. The combination of this technique and the altered medium promoted the migration and proliferation of SCs selectively by utilizing the supporting cells of SCs instead of discarding them by changing the culture dishes and media. PMID- 12573479 TI - The macrosphere model: evaluation of a new stroke model for permanent middle cerebral artery occlusion in rats. AB - BACKGROUND AND PURPOSE: The suture middle cerebral artery occlusion (MCAO) model is widely used for the simulation of focal cerebral ischemia in rats. This technique causes hypothalamic injury resulting in hyperthermia, which can worsen outcome and obscure neuroprotective effects. Herein, we introduce a new MCAO model that avoids these disadvantages. METHODS: Permanent MCAO was performed by intraarterial embolization using six TiO(2) macrospheres (0.3-0.4 mm in diameter) or by the suture occlusion technique. Body temperature was monitored, functional and histologic outcome was assessed after 24 h. Additional 16 rats were subjected to macrosphere or suture MCAO. Lesion progression was evaluated using magnetic resonance imaging (MRI). RESULTS: The animals subjected to suture MCAO developed hyperthermia (>39 degrees C), while the temperature remained normal in the macrosphere MCAO group. Infarct size, functional outcome and model failure rate were not significantly different between the groups. Lesion size on MRI increased within the first 90 min and remained unchanged thereafter in both groups. CONCLUSIONS: The macrosphere MCAO model provides reproducible focal cerebral ischemia, similar to the established suture technique, but avoids hypothalamic damage and hyperthermia. This model, therefore, may be more appropriate for the preclinical evaluation of neuroprotective therapies and can also be used for stroke studies under difficult conditions, e.g., in awake animals or inside the MRI scanner. PMID- 12573481 TI - Regulation of P450 genes by liver-enriched transcription factors and nuclear receptors. AB - Cytochrome P450s (P450s) constitute a superfamily of heme-proteins that play an important role in the activation of chemical carcinogens, detoxification of numerous xenobiotics as well as in the oxidative metabolism of endogenous compounds such as steroids, fatty acids, prostaglandins, and leukotrienes. In addition, some P450s have important roles in physiological processes, such as steroidogenesis and the maintenance of bile acid and cholesterol homeostasis. Given their importance, the molecular mechanisms of P450 gene regulation have been intensely studied. Direct interactions between transcription factors, including nuclear receptors, with the promoters of P450 genes represent one of the primary means by which the expression of these genes is controlled. In this review, several liver-enriched transcription factors that play a role in the tissue-specific, developmental, and temporal regulation of P450s are discussed. In addition, the nuclear receptors that play a role in the fine control of cholesterol and bile acid homeostasis, in part, through their modulation of specific P450s, are discussed. PMID- 12573482 TI - Functional and structural comparison of PXR and CAR. AB - The nuclear receptors pregnane X receptor (PXR, NR1I2) and constitutive active receptor (CAR, NR1I3) have both been proposed to function as xenosensors, but the details of their respective physiological roles are still being elucidated. We have contrasted these two receptors in a variety of experiments including gene expression assays, cell-based ligand profiling assays, and crystallographic/structural modeling analyses. These data highlight key differences between PXR and CAR. PMID- 12573483 TI - Phenobarbital induction of drug/steroid-metabolizing enzymes and nuclear receptor CAR. AB - Phenobarbital (PB) increases hepatic drug/steroid-metabolic capability by coordinately activating transcription of the genes encoding various metabolizing enzymes. The nuclear receptor CAR was first implicated as a transcription factor that activates the cytochrome P450 Cyp2b10 gene. In response to PB, CAR forms a heterodimer with the retinoid X receptor (RXR), binds to a PB response element (typified by DR-4 motif), and activates transcription of the gene. In the CAR null mouse, PB does not only induce the Cyp2b10 gene, but also induces genes encoding various metabolizing enzymes. Thus, CAR is a general nuclear receptor that is essential for PB induction of drug/steroid metabolizing enzymes. PB also induces amino levulinate synthase 1 (ALAS-1), the rate-limiting enzyme in heme biosynthesis, to increase heme supply. However, PB induction of the synthase occurs in CAR-null mice, suggesting that CAR does not coordinate the heme synthesis for the induction of drug/steroid metabolism. PMID- 12573484 TI - The expression of CYP2B6, CYP2C9 and CYP3A4 genes: a tangle of networks of nuclear and steroid receptors. AB - Numerous chemicals increase the metabolic capability of organisms by their ability to activate genes encoding various xenochemical-metabolizing enzymes, such as cytochromes P450 (CYPs), transferases and transporters. For example, natural and synthetic glucocorticoids (agonists and antagonists) as well as other clinically important drugs induce the hepatic CYP2B, CYP2C and CYP3A subfamilies in man, and these inductions might lead to clinically important drug-drug interactions. Only recently, the key cellular receptors that mediate such inductions have been identified. They include nuclear receptors, such as the constitutive androstane receptor (CAR, NR1I3), the retinoid X receptor (RXR, NR2B1), the pregnane X receptor (PXR, NR1I2), and the vitamin D receptor (VDR, NR1I1) and steroid receptors such as the glucocorticoid receptor (GR, NR3C1). There is a wide promiscuity of these receptors in the induction of CYPs in response to xenobiotics. Indeed, this adaptive system appears now as a tangle of networks, where receptors share partners, ligands, DNA response elements and target genes. Moreover, they influence mutually their relative expression. This review is focused on these different pathways controlling human CYP2B6, CYP2C9 and CYP3A4 gene expression, and the cross-talk between these pathways. PMID- 12573485 TI - Transcriptional analysis in vivo of the hepatic genes, Cyp2b9 and Cyp2b10, by intravenous administration of plasmid DNA in mice. AB - Phenobarbital (PB) responsiveness of CYP2B genes has been shown to be mediated by a PB responsive unit (PBRU). The core of the PBRU contains two nuclear receptor sites, NR-1 and NR-2, and a nuclear factor-1 (NF 1) binding site, which are required for PB responsiveness, but the importance of sequences flanking the core is not clear. We have used intravenous administration of plasmid DNA in the tail veins of mice to transfect hepatocytes in vivo and analyze sequence requirements for PB induction. In this assay PB treatment increased transactivation by the Cyp2b10 PBRU about 100-fold, which is similar to the increase in the expression of the endogenous gene while the Cyp2b9 PBRU was unresponsive. Analysis of chimeras of the two PBRUs and deletion mutants of the Cyp2b10 PBRU indicated that the core region containing the NR-1, NR-2 and NF-1 core sites is not sufficient for PB responsiveness. Additional sequence at the 3' side of the core sequence, which included a previously defined accessory factor-1 (AF-1) site, partially restored responsiveness. This region contained a binding site for NF-1 only in Cyp2b10 and not in Cyp2b9, but the intact site was not required for PB responsiveness. Purified constitutive androstane receptor (CAR)/retinoid X receptor (RXR) bound to the core NR-1 and NR-2 sites and to a third NR-3 site to the 5' side of the core in Cyp2b10. No binding of CAR/RXR to the Cyp2b9 PBRU was observed. These results indicate that changes in the NR sites which eliminate CAR/RXR binding are sufficient for the non-responsiveness to PB of Cyp2b9, but changes in sequences flanking the core independently eliminate PB responsiveness. The results demonstrate the advantages of transfection of mouse hepatocytes in vivo by tail vein injection of DNA as a method for transcriptional analysis of genes in vivo and show that sequences flanking the core region of the PBRU are required for PB induction in vivo. PMID- 12573486 TI - Functional role of AhR in the expression of toxic effects by TCDD. AB - Cytochrome P450 1A1 (CYP1A1) is one of the xenobiotic metabolizing enzymes (XMEs), which is induced by polycyclic aromatic hydrocarbons (PAHs). The most potent inducer of CYP1A1 is 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In addition, TCDD induces a broad spectrum of biochemical and toxic effects, such as teratogenesis, immunosuppression and tumor promotion. Most, if not all, of the effects caused by TCDD and other PAHs are known to be mediated by AhR (aryl hydrocarbon receptor or dioxin receptor) which has a high binding affinity to TCDD. The liganded AhR translocates from cytoplasm to nuclei where it switches its partner molecule from Hsp90 to Arnt. Thus formed AhR/Arnt heterodimer binds a specific DNA sequence designated XRE in the promoter region of the target genes including CYP1A1, UDP-glucuronosyl transferase and others to enhance their expression. Although it remains to be studied how AhR is involved in the other TCDD-induced biological effects such as teratogenesis and immunosuppression than induction of XMEs, it is believed that these adverse TCDD effects are caused by untimely activation of gene expression by ligand-activated AhR in the biological process. We summarize the present knowledge about functional role of AhR in TCDD induced biological effects. PMID- 12573487 TI - Transcriptional response elements in the promoter of CYP6B1, an insect P450 gene regulated by plant chemicals. AB - Papilio polyxenes, a lepidopteran continually exposed to toxic furanocoumarins in its hostplants, owes its tolerance to these compounds to the transcriptional induction of the CYP6B1 gene encoding a P450 capable of metabolizing linear furanocoumarins, such as xanthotoxin, at high rates. Transient expression of various lengths of wild-type and mutant CYP6B1v3 promoter in lepidopteran Sf9 cells defines a positive element (XRE-xan) from -136 to -119 required for both basal and xanthotoxin-inducible transcription and a negative element from -228 to -146 that represses basal transcription. Fusion of the CYP6B1v3 XRE-xan element to the Drosophila melanogaster Eip28/29 core promoter indicates that the XRE-xan functions in conjunction with its own core promoter but not with a heterologous core promoter. Sequence searches of the CYP6B1v3 proximal promoter region revealed a number of putative elements (XRE-AhR, ARE, OCT-1, EcRE, C/EBP, Inr) sharing sequence similarity with those in other regulated vertebrate and insect promoters. Mutation of TGAC nucleotides shared by the overlapping EcRE/ARE/XRE xan indicates that this sequence is essential for basal and regulated transcription of this gene. Mutagenesis in the non-overlapping region of the EcRE indicates it modulates basal transcription. These findings are incorporated into a working model for regulation of this toxin-inducible promoter. PMID- 12573488 TI - Induction of nicotine-metabolizing CYP2B1 by ethanol and ethanol-metabolizing CYP2E1 by nicotine: summary and implications. AB - Alcohol and tobacco are frequently co-abused. Increased alcohol use and alcoholism are associated with smoking, and vice versa. Functional and/or metabolic cross-tolerance may contribute to this occurrence. This review summarizes recent studies published from our laboratory focusing on metabolic aspects of tolerance, which demonstrate that in rat, subchronic, behaviourally relevant doses of ethanol induce hepatic nicotine-metabolizing cytochrome P450 (CYP) 2B1, and that subchronically administered nicotine, at behaviourally relevant doses, induces hepatic ethanol-metabolizing CYP2E1. Increased CYP2B1 protein, mRNA and CYP2B1-mediated nicotine metabolism was observed following ethanol treatments. CYP2E1 protein and activity were induced by nicotine, but no changes were seen in levels of CYP2E1 mRNA. These data indicate that metabolic cross-tolerance may occur between nicotine and ethanol, suggesting that nicotine use may increase the elimination of ethanol, and ethanol use may increase the elimination of nicotine. Other implications, such as altered pharmacology and toxicology of drugs metabolized by these enzymes, as well as changes in pro carcinogen and pro-toxin activation are also discussed. PMID- 12573489 TI - Molecular dynamics of substrate complexes with hamster cytochrome P450c17 (CYP17): mechanistic approach to understanding substrate binding and activities. AB - The cytochrome P450c17 isoforms from various animal species have different substrate selectivity, especially for 17,20-lyase activity. In particular, the human P450c17 selectively produces dehydroepiandrosterone with little androstenedione (AD). Hamster P450c17, on the other hand, produces both of these steroids at comparable rates. We thus investigated if computational analysis could explain the difference in activity profiles. Therefore, we inserted the four P450c17 substrates-pregnenolone, progesterone, and their 17alpha hydroxylated forms-inside our hamster P450c17 model, which we derived from our human P450c17 model based on the crystal structure of P450BMP. We performed molecular dynamics (MD) simulations on the complexes and analyzed the resultant trajectories to identify amino acids that interact with substrates. Starting with substrates in two different orientations, we obtained two sets of binding trajectories in each case. The first set of trajectories reveal structural rearrangements that occur during binding, whereas the second set of trajectories reflects substrate orientations during catalysis. Our modeling suggests that three distinct steps are required for substrate selectivity and binding to the hamster P450c17: (1) recognition of the substrate at the putative substrate entrance, characterized by a pocket at the surface of the hamster P450c17 containing charged residues R96 and D116; (2) entry of the substrate into the active site, in an intermediate position directed by possible hydrogen bonding of the substrates with the heme D-ring propionate group, R96, R440, and T306; followed by (3) 90 degrees counterclockwise rotation of the substrates, positioning them in optimal position for reactivity, a process that may be directed by hydrogen bonding to the 110-112 region of the hamster P450c17. With some substrates, we obtained trajectories which suggest that major distortions in the I-helix and opening of the H-I loop occur during substrate binding. In conclusion, these modeling exercises provide insight to possible structural reorganizations that occur during substrate binding and suggest that amino acids that participate in three distinct steps of this process may all contribute to substrate binding and activity. PMID- 12573490 TI - Hippocampal cytochrome P450s synthesize brain neurosteroids which are paracrine neuromodulators of synaptic signal transduction. AB - Hippocampal pyramidal neurons and granule neurons of adult male rats are equipped with a complete machinery for the synthesis of pregnenolone, dehydroepiandrosterone, 17beta-estradiol and testosterone as well as their sulfate esters. These brain neurosteroids are synthesized by cytochrome P450s (P450scc, P45017alpha and P450arom) from endogenous cholesterol. Synthesis is acutely dependent on the Ca(2+) influx attendant upon neuron-neuron communication via N-methyl-D-aspartate (NMDA) receptors. Pregnenolone sulfate, estradiol and corticosterone rapidly modulate neuronal signal transduction and the induction of long-term potentiation via NMDA receptors and putative membrane steroid receptors. Brain neurosteroids are therefore promising neuromodulators that may either activate or inactivate neuron-neuron communication, thereby mediating learning and memory in the hippocampus. PMID- 12573491 TI - The undifferentiated cell zone is a stem cell zone in adult rat adrenal cortex. AB - The adrenal cortex of mammals has been known to consist of three morphologically and functionally distinct zones, i.e. the zona glomerulosa (zG), the zona fasciculata (zF) and the zona reticularis (zR), each of which secretes a specific corticosteroid different from those produced by the other two zones. We found previously, however, that an additional zone existed between zG and zF of adult rat adrenal cortex and that the cells in that zone were in a functionally undifferentiated state as an adrenocortical cell [Endocrinology 135, (1994) 431]: they were incapable of synthesizing highly active forms of corticosteroids, such as aldosterone and corticosterone, although they could produce their precursors. Hence, we named the zone as the undifferentiated cell zone (zU) of the adrenal cortex. Here we show that zU and its surroundings, i.e. the innermost portion of zG and the outermost portion of zF are the sites for cell replication in adult rat adrenal cortex and that the cells raised there migrate to other regions. Such cell replications in this region occur regardless of physiological conditions, such as the rise and fall of hormonal stimuli and circadian fluctuation of adrenocortical activities. On the bases of these and other findings previously described, we propose that zU is the stem cell zone of the adult rat adrenal cortex. Our recent success in isolating novel cell lines, which display an undifferentiated phenotype similar to that of zU cells, could facilitate the exploration of molecular mechanisms for the differentiation and development of the adrenocortical cells. PMID- 12573492 TI - Expression of cytochromes P450 4F4 and 4F5 in infection and injury models of inflammation. AB - Lipopolysaccharide (LPS) treatment of rats suppresses CYP 4F4 and 4F5 expression by 50 and 40%, respectively, in a direct fashion occurring in the liver. This contention is borne out by essentially parallel dose-dependent changes observed upon treatment of rat hepatocyte cultures with LPS. An alternate avenue of triggering the inflammatory cascade is traumatic brain injury by controlled cortical impact. Such injury brings about a dramatic change in the expression of CYP 4F4 and 4F5 mRNA which reaches its greatest effect 24 h after impact compared with sham-operated but uninjured controls. At time points after 24 h the expression of both isoforms increases dramatically reaching highest levels at 2 weeks post-injury. These changes in mRNA expression are mirrored by changes in protein expression. The results are consistent with the notion that immediately after injury concentrations of leukotriene and prostaglandin mediators are elevated by decreased CYP 4F concentrations. As time after injury increases those conditions reverse. Increased CYP 4F expression leads to diminished concentrations of leukotriene and prostaglandin mediators and then to recovery and repair. PMID- 12573493 TI - Cytochrome P450 epoxygenases and vascular tone: novel role for HMG-CoA reductase inhibitors in the regulation of CYP 2C expression. AB - Over the last 10 years it has become increasingly clear that cytochrome P450 (CYP) enzymes expressed within endothelial and vascular smooth muscle cells play a crucial role in the modulation of vascular homeostasis. There is strong evidence suggesting that the activation of a CYP 2C epoxygenase in endothelial cells is an essential step in nitric oxide (NO)- and prostacyclin (PGI(2)) independent vasodilatation of several vascular beds, particularly in the heart and kidney. Moreover, CYP epoxygenase products as well as CYP-derived reactive oxygen species are intracellular signal transduction molecules involved in several signaling cascades affecting numerous cellular processes, including vascular cell proliferation and angiogenesis. Various pharmacological compounds enhance vascular CYP 2C expression. One group of substances which highlight the possible effects of CYP induction in endothelial cells on vascular function are the HMG-CoA reductase inhibitors (statins). Cerivastatin and fluvastatin increase CYP 2C mRNA and protein in native and cultured endothelial cells, and enhance the bradykinin-induced NO/PGI(2)-independent relaxation of arterial segments as well as the generation of reactive oxygen species. However, statins also increase the expression of the endothelial NO synthase by approximately twofold. As a consequence, the probability that NO and reactive oxygen species react to generate peroxynitrite is increased and the treatment of vascular segments with statins resulted in enhanced protein tyrosine nitration. These data highlight the role played by CYP 2C in vascular homeostasis and its potential regulation by cardiovascular drugs. PMID- 12573494 TI - Metapneumoviruses in birds and humans. AB - Avian pneumovirus (APV, Turkey rhinotracheitis virus) and Human metapneumovirus (hMPV) are pathogens of birds and humans, respectively, that are associated with upper respiratory tract infections. Based on their different genomic organization and low level of nucleotide (nt) and amino acid (aa) identity with paramyxoviruses in the genus Pneumovirus, APV and hMPV have been classified into a new genus referred to as Metapneumovirus. First isolated in 1970s, APV strains have since been isolated in Europe, Africa, middle east, and United States (US) and classified in four subgroups, APV/A, APV/B, APV/C, and APV/D based on nt and predicted aa sequence identity. Although it was first isolated in 2001, serological evidence indicates that hMPV may have been present in human population from as early as the 1950s. There is only one subgroup of hMPV so far, whose nt and aa sequence identity indicates that it is more closely related to APV/C than to APV/A, APV/B, or APV/D. PMID- 12573495 TI - Molecular analysis of the cucumber leaf spot virus genome. AB - Full-length clones of the genome of the Aureusvirus, Cucumber leaf spot virus (CLSV), have been constructed and infectious T7 polymerase derived synthetic transcripts have been produced. Mutational analysis of the genome indicates a role for p84 in viral RNA replication, the CP in systemic movement, p27 in viral cell-to-cell movement and p17 in symptom induction. A CLSV mutant lacking ORFs for the CP, p27 and p17 (CLSV YX) was capable of replication and systemic movement in transgenic Nicotiana benthamiana plants expressing the Red clover necrotic mosaic virus (RCNMV) movement protein (MP) suggesting that p25 and p84 are sufficient for viral RNA replication and that the RCNMV MP can permit CLSV cell-to-cell as well as systemic movement. Moreover, CLSV YX induced severe necrosis in both inoculated and uninoculated leaves of transgenic plants suggesting that CLSV p25 and/or p84 are important determinants of the necrotic phenotype. Another mutant similar to CLSV YX but expressing only limited amino terminal portions of CP, p27 and p17 failed to produce necrosis or to move systemically in RCNMV MP transgenic N. benthamiana plants. These results suggest that these short translated regions or cis-acting sequences present in the CLSV CP, p27 and/or p17 ORFs suppress the necrosis induced by p25/p84 and also suppress systemic movement mediated by the RCNMV MP. PMID- 12573496 TI - Differential stability and fusion activity of Lyssavirus glycoprotein trimers. AB - The oligomeric structure and the fusion activity of lyssavirus glycoprotein (G) was studied by comparing G from Mokola virus (GMok) and rabies virus (PV strain) (GPV), which are highly divergent lyssaviruses. G expressed at the surface of BSR cells upon either plasmid transfection or virus infection are shown to be mainly trimeric after cross-linking experiments. However, solubilization by a detergent (CHAPS) and analysis in sucrose sedimentation gradient evidenced that GMok trimer is less stable than GPV trimer. A chimeric glycoprotein (G Mok-PV) associating the N-terminal half of GMok to the C-terminal half part of GPV formed trimers with an intermediate stability, indicating that the G C-terminal domain is essential in trimer stability. A cell to cell fusion assay revealed that GMok (and not G Mok-PV) was able to induce fusion at a higher pH (0.5 pH unit) than GPV. Such differences in the oligomeric structure stability and in the fusion activity of lyssavirus glycoproteins may partly account for the previously reported differences of their immunogenic and pathogenic properties. PMID- 12573497 TI - Nucleotide sequence analysis of the accessory genes of HIV-1 group O isolates. AB - Human immunodeficiency virus type 1 group O strains have been described as highly divergent, compared with the majority of the viruses classified in group M. To study the diversity and genetic characteristics of group O, we have sequenced the accessory genes of 7 isolates. Analysis of the deduced amino acid sequences for Vif, Vpr, Tat, Vpu, and Rev indicate that most of the functional domains of these proteins, as described for group M viruses, are highly conserved and retained among all the group O strains we have characterized. The only difference concerns the Vif phosphorylation sites, which are absent in all of the group O isolated we have sequence with the exception of two isolates in which only one phosphorylation site was conserved. These sites, present in nearly all of the group M isolates, play a critical role in the regulation of viral replication and infectivity. As described for group M isolates, the vpu gene is the one with the highest diversity among group O viruses. Phylogenetic analysis of these sequences suggests that group O viruses could be differentiated into at least four different clusters. PMID- 12573498 TI - Complete genomic sequences of astroviruses from sheep and turkey: comparison with related viruses. AB - The genomes of astroviruses infecting sheep and turkey were sequenced. Detailed analyses of these sequences were performed, including comparison with the other complete astrovirus sequences available as well as with other RNA virus sequences, with focus on the non-structural proteins and RNA sequences. Earlier postulated functional astrovirus RNA motifs and protein domains could in most cases be recognised in the sheep and turkey astrovirus sequences. In addition, analyses of the available astrovirus sequences revealed: two protein regions with the potential for forming coiled coils, differences in the postulated transmembrane region, a similarity between the putative astrovirus nuclear localisation signal and calicivirus genome-linked proteins, and a stretch of a highly conserved RNA sequence with a possible role in the astrovirus capsid gene expression. The present analyses contribute to the deciphering of pertinent functions of the astrovirus genomes. PMID- 12573499 TI - Molecular epidemiology of canid rabies in Zimbabwe and South Africa. AB - The epidemiology of rabies in southern Africa is complex, due to a large number of vector species and the presence of at least two distinct biotypes of the virus. Our objective was to contribute to the understanding of the epidemiology of rabies in the southern African subcontinent by studying the genetic relationship of 89 rabies virus isolates from this region. In this study, we have focused on an analysis of viruses that cycle in canid host species (canid biotype) throughout South Africa and Zimbabwe. By phylogenetic analysis of the cytoplasmic domain of the glycoprotein and the non-coding G-L intergenic region, all the southern African canid viruses were found to be closely related and no apparent general distinction could be made between them. Although there was a minor degree of phylogenetic branching, with certain branches associated with cycles defined by species, location and time, the phylogenetic pattern indicated that canid rabies in southern Africa is derived from a single virus lineage, which has spread opportunistically within whatever canid host population is ecologically capable of sustaining prolonged cycles. This molecular epidemiological study presents the first comprehensive comparison of rabies viruses from South Africa and Zimbabwe and has demonstrated the need for multinational approaches towards the control of this important zoonotic disease in Africa. PMID- 12573500 TI - Studies of measles viruses circulating in Australia between 1999 and 2001 reveals a new genotype. AB - Nineteen distinct measles virus (MV) strains associated with nine different genotypes were identified in five Australian states (Victoria, New South Wales, Queensland, Northern Territory and Western Australia) between 1999 and 2001. One of the strains identified is likely to represent a new genotype within the clade D viruses (proposed to be d9). No evidence for an indigenous MV strain was found. When epidemiologic information associated with the index case was available for the outbreaks, it usually supported introduction of the virus from overseas, with the main source being South East Asia. Changes in the circulation of MV in Australia since the early 1970s were also observed. Prior to the introduction of measles vaccine, the majority of the population acquired immunity through infection with wild-type virus in early childhood. Nowadays in Australia, young adults are at most risk of infection. The age range of cases in the study period was from 1 month to 48 years, with the majority (59%) of cases from individuals aged 18-30 years. PMID- 12573501 TI - Adaptable modification of adenoviral tropism using a bifunctional ligand protein. AB - In order to target recombinant adenovirus (AdV), we have developed a new strategy using a fusion ligand protein comprising coxsackievirus/adenovirus receptor (CAR), and the antibody Fc-binding domain from protein A in vitro testing with this ligand shows that it blocks viral gene transduction and, when coupled with anti-ICAM-1 IgG, redirects AdV to endothelial cells that are induced to express ICAM-1. Because the protein A Fc-binding domain will bind to any immunoglobulin, the current strategy can be adapted to target a wide variety of tissues or cells as long as an antibody species that recognizes a membrane marker on target tissue or cell is present. This concept may be further expanded to other viruses that employ peptide receptors. These membrane receptors can be fused to the Fc-binding domain to create a variety of bifunctional ligands for targeting recombinant viruses in gene therapy. PMID- 12573502 TI - Defective human T-cell lymphotropic virus type I (HTLV-I) provirus in seronegative tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM) patients. AB - Infection with human T-cell lymphotropic virus type I (HTLV-I) have been associated with the development of the tropical spastic paraparesis/HTLV-I associated myelopathy (TSP/HAM). We studied the presence of HTLV-I provirus in peripheral blood mononuclear cells (PBMC) from 72 Chilean patients with progressive spastic paraparesis by polymerase chain reaction: 32 seropositive and 40 seronegative cases. We amplified different genomic regions of HTLV-I using primers of 5' ltr, tax, env/tax, pX, pol and env genes. These genes were detected from all seropositive patients. The seronegative patients were negative with 5' ltr, pol, env, and pX primers. However, amplified product of tax and env/tax genes was detected from 16 and four seronegative patients, respectively. Three of them were positive with both genetic regions. The results of this study show that the complete HTLV-I provirus is found in 100% of seropositive cases. In seronegative cases, clinically very similar of seropositive cases, was found only tax gene in 42.5% (17/40) of patients. These results suggest the presence of a defective HTLV-I provirus in some seronegative patients with progressive spastic paraparesis, and suggest a pathogenic role of this truncate provirus for a group of TSP/HAM. PMID- 12573503 TI - Molecular epidemiology and type-specific detection of echovirus 11 isolates from the Americas, Europe, Africa, Australia, southern Asia and the Middle East. AB - Echovirus 11 (E11) is among the most commonly isolated human enteroviruses. To examine the range of genetic variation within the E11 serotype, we determined the complete VP1 sequences for 53 geographically dispersed E11 strains isolated in 16 countries from 1953 to 2001. E11 sequences were monophyletic with respect to all other enterovirus serotypes. The sequences clustered into four monophyletic genogroups, A-D; members of each genogroup differed from one another by <20%. Isolates in different genogroups differed from one another by 19-28%. The E11 prototype strain, USA/CA53-Gregory, was the sole member of genogroup B. All recent US isolates were members of one of two discrete lineages within genogroup D. The well-characterized E11 antigenic variant, USA/CA63-Silva, was also a member of genogroup D. Members of genogroups A and C were antigenically similar to USA/CA53-Gregory, as measured by neutralization with anti-Gregory and anti Silva antisera. Only USA/CA63-Silva was neutralized more efficiently by the anti Silva antiserum; other genogroup D viruses were Gregory-like or intermediate in their neutralization phenotype. Recent non-US isolates were distributed in genogroups A, C and D. Sequence similarities among genogroup D isolates from North America, Europe, Asia, Australia and North Africa demonstrate that an E11 strain can spread rapidly over a wide geographic area. The aligned sequences were used to develop an E11-specific RT-PCR assay, using degenerate, inosine containing primers, to amplify all members of all genogroups. PMID- 12573504 TI - Sensitivity of Invertebrate iridescent virus 6 to organic solvents, detergents, enzymes and temperature treatment. AB - The sensitivity of Invertebrate iridescent virus 6 (IIV-6) to a selection of organic solvents, detergents, enzymes and heat treatment was assayed in Spodoptera frugiperda (Sf9) cells and by injection of inoculum into larvae of Galleria mellonella. In several cases, the degree of sensitivity of the virus depended on the method of assay; cell culture assays indicated greater losses of activity than insect bioassay. IIV-6 was sensitive to chloroform but sensitivity to ether was only detected by cell culture assay. Sensitivity (defined as a reduction of at least 1 log activity) was detected following treatment by 1 and 0.1% SDS, 1% Triton-X100, 70% ethanol, 70% methanol, 1% sodium deoxycholate, pH 11.1 and 3.0. No sensitivity was detected to 1% Tween 80, 1 M MgCl2, 100 mM EDTA, lipase, phospholipase A2, proteinase K, or trypsin at the concentrations tested. Viral activity was reduced by approximately 4 logs following heating to 70 degrees C for 60 min or 80 degrees C for 30 min. The above observations highlight the need for studies on the role of the virus lipid component in the process of particle entry into cells, and may explain why vertebrate and invertebrate iridoviruses have been reported to differ in their sensitivity to organic solvents and enzymes. PMID- 12573505 TI - Cytotoxic T-cell activity is not detectable in Venezuelan equine encephalitis virus-infected mice. AB - Previously published research has established that the immune response to the Venezuelan equine encephalitis virus (VEEV) vaccine strain TC-83 is Th 1 mediated, with local activation of both CD4+ and CD8+ T cells. This suggests that cytotoxic lymphocytes CTL may play a role in protection against virulent VEEV. Studies involving a variety of immunisation schedules with either TC-83 or strain CAAR 508 (serogroup 5) of VEEV, and six different haplotypes of mice, failed to reveal functional CTL activity against VEEV-infected targets in secondary antigen stimulated lymphocyte cultures from either the draining lymph nodes (LN) or spleen. Nor were VEEV-specific CTL detected after immunisation of mice (three haplotypes) with recombinant vaccinia viruses (VV) expressing either the non structural (nsP1-4) or the structural (C-E3-E2-6K-E1) genes of TC-83. Reciprocal experiments in which mice were immunised with TC-83, and their lymphocytes tested against VV recombinant-infected targets also failed to detect CTL activity. These data suggest that VEEV infection of mice does not elicit detectable CTL activity, and that CTL are unlikely to play a role in protection against virulent VEEV. PMID- 12573507 TI - Pharmacology of ischemia-induced glutamate efflux from rat cerebral cortex in vitro. AB - Simulated ischemic conditions (hypoxia-hypoglycaemia) in vitro enhanced glutamate efflux from rat cerebrocortical prisms. Here we characterised efflux mechanisms using pharmacological tools. The Na(+) channel blocker TTX (1 microM) did not affect ischemia-induced efflux, while sipatrigine (100 microM), a Na(+)/Ca(2+) channel blocker and omega-conotoxin MVIIC (2 microM), an N/P/Q type Ca(2+) channel blocker, inhibited efflux by fractions of 0.53 and 0.46, respectively (1.00 corresponding to total inhibition). Omission of extracellular Ca(2+) and addition of EGTA (2 mM) inhibited ischemia-induced efflux only during the first 25 min of incubation. A similar result was observed on omission of extracellular Ca(2+) together with addition of La(3+) (10 microM) and Mg(2+) (6 mM). TTX, sipatrigine and La(3+)/Mg(2+) all inhibited control efflux. Ischemia-induced efflux was sensitive to the volume activated anion channel inhibitor NPPB (100 microM) only after the first 25 min of incubation, with the maximal fraction inhibited being 0.54. The glutamate transporter inhibitor D,L-TBOA reduced ischemia-induced efflux throughout a 45-min incubation period, and enhanced efflux from control tissue. D,L-TBOA inhibited efflux at 30 min by a maximum fraction of 0.49, at 50 microM. These data indicate that the early phase of ischemia-induced glutamate efflux is in part Ca(2+) dependent, while the later phase involves volume activated anion currents and both phases involve excitatory amino acid transporters. PMID- 12573508 TI - Non-adrenergic exploratory behavior induced by moxonidine at mildly hypotensive doses. AB - Moxonidine is a centrally-active imidazoline compound with preferential affinity for imidazoline receptors (IR) over alpha(2)-adrenoceptors (alpha(2)AR). Clinically, moxonidine has proven advantageous for treating hypertension over pure alpha(2)-adrenergic agonists (i.e., guanabenz) due to its lowered incidence of sedative side effects. The present experiments reveal divergent behavioral effects of low doses of moxonidine and guanabenz in C57Bl/6 mice in an exploratory arena. Low-dose moxonidine (0.05 mg kg(-1) i.p.) elicited an increase in novel object contacts (+36%) and more movement into central space (+56%; P<0.01) compared to saline-injected controls; whereas guanabenz induced only dose responsive sedative-like behaviors in the same paradigm. Yet, the two agonists were indistinguishable in terms of blood pressure changes over a similar dose range (0.025-0.1 mg kg(-1) i.p.) in consciously free-moving mice (Delta mean+/ S.E.M.=-12.3+/-3.2 mm Hg for moxonidine versus -13.5+/-1.9 mm Hg for guanabenz). As expected of alpha(2)AR involvement, the sedative-like effects of guanabenz were completely blocked by pretreatment with the non-imidazoline alpha(2)AR antagonist, SKF86466 (0.5 or 1.0 mg kg(-1) i.p.). However, the pro-exploratory effects of low doses of moxonidine (0.05 or 0.1 mg kg(-1)) were not antagonized by SKF86466. These results suggest that moxonidine acts preferentially through a non-adrenergic mechanism, possibly IR-mediated, to elicit pro-exploratory behavior. PMID- 12573509 TI - Choline acetyltransferase is expressed by non-starburst amacrine cells in the ground squirrel retina. AB - We have used immunostaining techniques to reveal a new type of amacrine cell that is immunoreactive for choline acetyltransferase (ChAT), the acetylcholine synthesizing enzyme, in the Ground Squirrel (Spermophilus beecheyi) retina. Cryostat sections and double immunostained wholemount preparations were examined by confocal microscopy. This new ChAT type III cell is distinct in morphology and neurotransmitter content from the well know 'starburst' amacrine cells (types I and II) that are so well represented in the ground squirrel retina [J. Comp. Neurol. 365 (1996) 173-216]. The type III cell colocalizes glycine with the acetylcholine and does not appear to be GABAergic or exhibit calcium-binding proteins like the well-known starburst type. As well, type III cells do not occur as a mirror-symmetric pair with normally placed and displaced varieties. The type III cell is probably a small field amacrine type branching broadly in upper sublamina b of the inner plexiform layer, and is most likely A6 of the Ground Squirrel retina [J. Comp. Neurol. 365 (1996) 173-216]. Type III cells are ideally placed in the architecture of the Ground Squirrel retina to influence ON directionally selective ganglion cell types. PMID- 12573510 TI - Superior colliculus stimulation enhances neocortical serotonin release and electrocorticographic activation in the urethane-anesthetized rat. AB - Recent evidence indicates that the superior colliculus (SC), in addition to its functions in sensory detection, also participates in controlling the generalized activation state of the forebrain, as measured by the electroencephalogram (EEG) or electrocorticogram (ECoG). The mechanisms by which the SC modulates forebrain activation are not well understood. By using in vivo microdialysis, we examined the role of serotonin release as a mechanism by which the SC can control neocortical activity in the urethane-anesthetized rat. Electrical 100 Hz stimulation of the SC increased frontal cortex serotonin output to 116, 118, and 140% of baseline levels for stimulation intensities of 0.5, 0.75, and 1.0 mA, respectively. Further, 75% of extracellularly recorded single (putative serotonergic) dorsal raphe neurons increased their discharge rate in response to 100 Hz stimulation of the SC. Stimulation of the SC also suppressed frontal cortex low frequency (1-6 Hz) synchronized ECoG activity, replacing it with high frequency desynchronization. This activation response was resistant to cholinergic-muscarinic receptor antagonists (atropine, 50 mg/kg; scopolamine, 2 mg/kg), but was reduced or abolished by systemic treatment with the serotonergic receptor antagonists ketanserin (10 mg/kg) or methiothepin (5 mg/kg). These data suggest that efferents from the SC, possibly by an excitatory action on serotonergic dorsal raphe cells, produce an enhanced release of serotonin and ECoG activation in the neocortex. The stimulation of cortical serotonin output may constitute a mechanism by which the SC acts on the forebrain to increase cortical excitability in response to sensory stimuli processed by SC neurons. PMID- 12573511 TI - The effects of prostaglandin E2 on the firing rate activity of thermosensitive and temperature insensitive neurons in the ventromedial preoptic area of the rat hypothalamus. AB - In response to an immune system challenge with lipopolysaccharide (LPS), recent work has shown that Fos immunoreactivity is displayed by neurons in the ventromedial preoptic area of the hypothalamus (VMPO). In addition, neurons in this region show distinct axonal projections to the anterior perifornical area (APFx) and the paraventricular nucleus (PVN). It has been hypothesized that neurons within the VMPO integrate their local responses to temperature with changes in firing activity that result from LPS induced production of prostaglandin E(2) (PGE(2)). This may be an important mechanism by which the set point regulation of thermoeffector neurons in the APFx and PVN is altered, resulting in hyperthermia. To characterize the firing rate activity of VMPO neurons, single-unit recordings were made of neuronal extracellular activity in rat hypothalamic tissue slices. Based on the slope of firing rate as a function of tissue temperature, neurons were classified as either warm sensitive or temperature insensitive. Neurons were then treated with PGE(2) (200 nM) while tissue temperature was held at a constant level ( approximately 36 degrees C). The majority of temperature insensitive neurons responded to PGE(2) with an increase in firing rate activity, while warm sensitive neurons showed a reduction in firing rate. This suggests that both warm sensitive and temperature insensitive neurons in the VMPO may play critical and contrasting roles in the production of a fever during an acute phase response to infection. PMID- 12573512 TI - The effect of taste stimuli on histamine release in the anterior hypothalamus of rats. AB - We studied the effect of gustatory stimulation on hypothalamic histamine release. Administering a four-basic taste mixture significantly increased histamine release, but not in the chorda tympani-transected rats. 0.1 M NaCl significantly increased histamine release, whereas 0.5 M sucrose, 0.02 M quinine HCl and 0.01 M HCl had no effect. However, when the concentration of HCl was increased to 0.03 M, a significant increase in histamine release was seen. These results suggest that taste information via the chorda tympani activates the histaminergic system. PMID- 12573513 TI - Enhancement of BDNF and activated-ERK immunoreactivity in spinal motor neurons after peripheral administration of BDNF. AB - Brain-derived neurotrophic factor (BDNF) shows neurotrophic effects on adult motor neurons when given systemically, But it is unknown whether systemically administered BDNF is transported to central cell bodies to affect them directly. Here we used immunohistochemistry to investigate the transport of peripherally injected BDNF to spinal motor neurons and the subsequent activation of a signaling pathway. We first injected BDNF into the flexor digitorum brevis (FDB) and analyzed the motor nucleus that projects to the FDB for BDNF immunoreactivity (BDNF-ir) and phosphorylated extracellular signal-regulated kinase (ERK) 1/2 immunoreactivity (pERK1/2-ir). Both immunoreactivities were observed in the motor neuron cell bodies. Next, BDNF was injected subcutaneously (s.c.) into rats with a unilaterally axotomized sciatic nerve. pERK1/2-ir was detected in motor neurons of the lesioned side. BDNF-ir and pERK1/2-ir were also observed on the unlesioned side when a high dose of BDNF was injected. Therefore, we examined BDNF-ir and pERK1/2-ir after injecting BDNF s.c. into normal rats. Both immunoreactivities were observed in motor nuclei on both sides. Finally, we examined pERK1/2-ir after a lower dose of BDNF was injected, which prevents the decrease in choline acetyl transferase that occurs in the motor neuron upon axotomy. Spinal motor nuclei contained a few cell bodies with pERK1/2-ir. These findings represent the first direct evidence that subcutaneously injected BDNF is transported to motor neurons and that it activates a signaling pathway in the spinal cord and exhibits neurotrophic effects in vivo. PMID- 12573514 TI - Modulation of monosynaptic transmission by presynaptic inhibition during fictive locomotion in the cat. AB - The effect of multisensory inputs onto the presynaptic inhibitory pathways affecting IA terminals was studied during fictive locomotion in decerebrated cats. The effect was evaluated from changes in amplitude of the monosynaptic excitatory postsynaptic potential (EPSP) measured in lumbosacral motoneurones. Responses were grouped and averaged according to their timing within the step cycle divided into five bins. Presynaptic inhibition was evoked by stimulating group I afferents from the posterior biceps-semitendinosus (PBSt) muscles and one of three cutaneous nerves: superficial peroneal (SP), sural and saphenous. Statistical analysis was applied to compare (1) EPSPs conditioned by PBSt input alone and those conditioned by the combined PBSt and cutaneous inputs, and (2) each bin dividing the step cycle to disclose phase-dependent changes. Results from 19 motoneurones showed that: (1) there was a significant phase-dependent modulation in EPSP amplitude (by 25%) with the maximum usually occurring during the depolarized phase; (2) PBSt alone reduced the EPSP amplitude (by 21%) in 3.2 bins on average; (3) combined PBSt and cutaneous stimuli further modified (up or down) the EPSP amplitude in half the trials but only in one to two bins; and (4) the most efficient cutaneous nerve (SP) usually decreased the PBSt-evoked reduction in EPSP size. Minimal changes in membrane input resistance suggest that the EPSP modifications were mostly due to presynaptic inhibition. Results indicate that muscle afferents can induce an important phase-dependent presynaptic inhibition of monosynaptic transmission and that concomitant activation of cutaneous afferents can alter this inhibition but only for a restricted part of the step cycle. PMID- 12573515 TI - Chronic inhibition of the norepinephrine transporter in the brain participates in seizure sensitization to cocaine and local anesthetics. AB - The involvement of chronic inhibition of monoamine transporters (MAT) in the brain with respect to sensitization to cocaine- and local anesthetic-induced seizures was studied in mice. Repeated administration of subconvulsive doses of meprylcaine as well as cocaine, both of which inhibit MAT, but not lidocaine, which does not inhibit MAT, increased seizure activity and produced sensitization to other local anesthetics. The effects of five daily treatments of monoamine transporter inhibitors on lidocaine-induced convulsions were examined 2 or 3 days after the last dose of the inhibitors. Daily treatments of GBR 12935, a specific inhibitor of dopamine uptake, significantly increased the incidence and the intensity of lidocaine-induced convulsions at 20 mg/kg and decreased the threshold of the convulsions. Daily treatments of desipramine and maprotiline, selective norepinephrine uptake inhibitors, markedly increased the incidence and intensity of lidocaine-induced convulsions, and decreased the threshold in a dose dependent manner at between 5 and 20 mg/kg. Daily treatments of citalopram, a selective serotonin uptake inhibitor, at 10 and 20 mg/kg, produced no significant increase in the incidence or intensity of lidocaine-induced convulsions, but decreased the threshold of the convulsions. These results suggest that the chronic intermittent inhibition of monoamine uptake increases susceptibility to cocaine- and local anesthetic-induced seizures, and the norepinephrine transporter is an integral component of this sensitization. PMID- 12573516 TI - Compartmentation of alpha 1 and alpha 2 GABA(A) receptor subunits within rat extended amygdala: implications for benzodiazepine action. AB - The extended amygdala, a morphological and functional entity within the basal forebrain, is a neuronal substrate for emotional states like fear and anxiety. Anxiety disorders are commonly treated by benzodiazepines that mediate their action via GABA(A) receptors. The binding properties and action of benzodiazepines depend on the alpha-subunit profile of the hetero-pentameric receptors: whereas the alpha1 subunit is associated with benzodiazepine type I pharmacology and reportedly mediates sedative as well as amnesic actions of benzodiazepines, the alpha2 subunit confers benzodiazepine type II pharmacology and mediates the anxiolytic actions of benzodiazepines. We determined the localization of alpha1 and alpha2 subunits within the extended amygdala, identified by secretoneurin immunostaining, to define the morphological substrates for the diverse benzodiazepine actions. A moderate expression of the alpha1 subunit could be detected in compartments of the medial subdivision and a strong expression of the alpha2 subunit throughout the central subdivision. It is concluded that the alpha1 and alpha2 subunits are differentially expressed within the extended amygdala, indicating that this structure is compartmentalized with respect to function and benzodiazepine action. PMID- 12573517 TI - Ganglioside GT1B and melatonin inhibit brain mitochondrial DNA damage and seizures induced by kainic acid in mice. AB - The effects of ganglioside GT1b or melatonin on damage to brain mitochondrial DNA (mtDNA) and seizures induced by kainic acid were investigated both in vivo and in vitro. An intraperitoneal (i.p.) injection of kainic acid (45 mg/kg) produced broad-spectrum limbic and severe sustained seizures in all of the treated mice. These seizures were completely abolished by an intracerebroventricular (i.c.v.) injection of ganglioside GT1b (90 nmol/brain), a potent inhibitor of glutamate receptor mediated activation and translocation of protein kinase C and lipid peroxidation, or an i.p. injection of melatonin (20 mg/kg), a potent scavenger of hydroxyl radicals (*OH). The administration of kainic acid caused damage to mtDNA in brain frontal and central portion of cortex in mice. The damage to mtDNA was abolished by pre-injection of ganglioside GT1b (90 nmol/brain, i.c.v.) or melatonin (20 mg/kg, i.p.). In vitro exposure of kainic acid (0.25, 0.5 or 1.0 mM) inflicted damage to mtDNA in a concentration-dependent manner. The damage to mtDNA induced by 1.0 mM kainic acid was attenuated by the co-treatment with 60 microM ganglioside GT1b or 1.5 mM melatonin. Furthermore, kainic acid (0.5 or 1.0 mM) increased lipid peroxidation in a concentration-dependent manner when incubated with a homogenate prepared from mice brain at 37 degrees C for 20 or 60 min. However, the increased lipid peroxidation was completely abolished by the co treatment with ganglioside GT1b (60 microM) or melatonin (1.5 mM). These results suggest that reactive oxygen species including hydroxyl radical (*OH) may play a role in the damage to brain mtDNA and seizures induced by kainic acid. We conclude that the preventive effect of melatonin or ganglioside GT1b against kainic acid-induced mtDNA damage or seizures may be due to its scavenging of reactive oxygen species including the *OH. PMID- 12573518 TI - Expression of receptors for insulin and leptin in the ventral tegmental area/substantia nigra (VTA/SN) of the rat. AB - Recent studies have demonstrated that the metabolic hormones insulin and leptin can modulate behavioral performance in reward-related paradigms. However, specific anatomical substrate(s) within the CNS for these effects remain to be identified. We hypothesize that midbrain dopamine neurons, which have been implicated to be critical in the mediation of motivational and reward aspects of stimuli, contribute to these behavioral effects of insulin and leptin. As one approach to evaluate this hypothesis, we used double-labeling fluorescence immunohistochemistry to determine whether the midbrain dopamine neurons express insulin receptors or leptin receptors. Extensive co-expression of tyrosine hydroxylase (a marker for dopamine neurons) with both the insulin receptor and the leptin receptor was observed in the ventral tegmentum and substantia nigra. These findings suggest that midbrain dopamine neurons are direct targets of insulin and leptin, and that they participate in mediating the effects of these hormones on reward-seeking behavior. PMID- 12573519 TI - Ability of NMDA and non-NMDA receptor antagonists to inhibit cerebral ischemic damage in aged rats. AB - Although stroke is a major cause of death and disability in the elderly, the inhibitory effects of neuroprotectants in acute stroke have been investigated using experimental cerebral ischemic models of young animals. Recent clinical trials have found that few neuroprotectants are effective. These observations indicate that effects in the clinical setting do not always reflect data from young animals. Thus, we compared the effects of the NMDA receptor antagonist MK 801 and of the AMPA receptor antagonist NBQX [2,3-dihydroxy-6-nitro-7-sulfamoyl benzo(F)quinixaline] on ischemic cerebral damage in the photothrombosis model of aged and young rats. MK-801 administered immediately after MCA occlusion significantly (P<0.05) reduced the extent of cerebral damage in young, but not in aged, rats and the effects of NBQX were similar. In separate experiments, we evaluated brain damage after microinjecting NMDA or kainic acid into the cortex using a stereotaxic apparatus. We found no significant differences in focal cerebral damage caused by NMDA between young and aged rats. On the other hand, kainic acid caused all of the aged rats tested to die, but none of the young rats. Our observations indicate that NMDA and AMPA receptor antagonists are less effective in aged, than in young, rats and that cerebral damage by receptor agonists depends on the type of receptor, such as NMDA and AMPA. PMID- 12573520 TI - Enriched environment delays the onset of hippocampal damage after global cerebral ischemia in rats. AB - An enriched environment has been shown to improve cognitive, behavioral and histopathological outcome after focal cerebral ischemia and head trauma. The purpose of this study was to determine the effect of an enriched environment on histopathology following global cerebral ischemia. Wistar rats (21 weeks of age) were placed in different environments [standard cages (SC) or enriched environment (EE) cages] for 2 months before and either 6 days or 2 months after ischemia. Rats underwent 10 min of global ischemia by bilateral carotid artery occlusions plus hypotension. Five groups (n=4-5 in each group) were studied: (1) rats kept in SC before and 2 months after ischemia; (2) rats kept in SC before ischemia but transferred to an EE for 2 months after ischemia; (3) rats kept in EE before and after ischemia for 2 months; (4) rats kept in SC before and 6 days after ischemia; (5) rats kept in EE before and 6 days after ischemia. At 7 days or 2 months after ischemia, brains were perfusion-fixed, and ischemic injury was assessed by counting numbers of normal neurons in the hippocampal CA1 sector. Physiological variables showed no inter-group differences. Rats housed in EE for 2 months before and for 6 days (but not 2 months) after global ischemia showed significantly better preservation of pyramidal neurons in the hippocampal CA1 area when compared to control animals (middle CA1, 20.5+/-5.4 vs. 2.8+/-0.6; lateral CA1, 31.5+/-7.2 vs. 2.6+/-0.6, respectively). The present data suggest that housing in EE for 2 months before and 6 days after ischemia can delay the onset of damage to hippocampal pyramidal neurons, which eventually occurs despite 2-month EE. PMID- 12573521 TI - Diabetes-induced neuroendocrine changes in rats: role of brain monoamines, insulin and leptin. AB - Diabetes is characterized by hyperphagia, polydypsia and activation of the HPA axis. However, the mechanisms by which diabetes produces these effects are not clear. This study was conducted to examine the effects of diabetes on the neuroendocrine system and to see if treatment with insulin and/or leptin is capable of reversing these effects. Streptozotocin-induced diabetic adult male rats were subjected to the following treatments: vehicle, insulin (2 U/day, s.c.), leptin (100 microg/kg BW) or leptin+insulin every day for 2 weeks. Food intake, water intake, and body weight were monitored daily. We measured changes in monoamine concentrations in discrete nuclei of the hypothalamus at the end of treatment. Diabetes produced a marked increase in food intake and water intake and this effect was completely reversed by insulin treatment and partially reversed by leptin treatment (P<0.05). Diabetes caused an increase in norepinephrine (NE) concentrations in the paraventricular nucleus with a concurrent increase in serum corticosterone. Treatment with insulin and leptin completely reversed these effects. Induction of diabetes also increased the concentrations of NE, dopamine and serotonin in the arcuate nucleus and NE concentrations in the lateral hypothalamus, ventromedial hypothalamus (VMH) and suprachiasmatic nucleus (P<0.05). Although insulin treatment was capable of reversing all these changes, leptin treatment was unable to decrease diabetes induced increase in NE concentrations in the VMH. These data provide evidence that hypothalamic monoamines could mediate the neuroendocrine effects of diabetes and that insulin and leptin act as important signals in this process. PMID- 12573522 TI - Possible regulatory mechanism of DHA-induced anti-stress reaction in rats. AB - To determine whether docosahexaenoic acid (DHA) affects stress responses in rats, we investigated its influence on several behavioral tests. Female rats were fed a diet deficient in (n-3) fatty acid from mating through pregnancy and lactation. Male pups fed the same diet as their dams were used for experiments. The effects of dietary (n-3) fatty acid deficiency and supplementation with DHA on psychological stress and conditioned-fear stress were investigated. The effect of DHA on psychological stress was examined by an elevated plus-maze test. The (n-3) deficient rats spent significantly (P<0.05) less time in the open arms; after 1 week of supplementation with DHA, they showed a significant (P<0.01) improvement. We then examined the paired effects of DHA and CRH on stress manifestations by an intracerebroventricular (i.c.v.) cannulation and behavior testing. An i.c.v. infusion of CRH (500 pmol) under resting conditions was shown to have stress inducing effects on behavior such as decreases of rearing, smelling and feeding, and increases of face washing; the supplementation of DHA significantly improved these distress behaviors. Finally, conditioned fear was induced by 40 min forced exposure to a cage in which the rat had experienced footshocks (30 x 1 mA x 1 s) 1 day before. Freezing behavior was dramatically suppressed by the supplementation of DHA, even 48 h after the conditioning treatment. Furthermore, the effect of DHA on the conditioned fear stress response is maintained over a long-term period. The i.c.v. pre-treatment of rats with bicuculline, a GABA(A) receptor antagonist, enhanced the conditioned-fear-induced freezing time in a dose-dependent fashion in the (n-3) fatty acid deficient animals. Significantly, the DHA supplemented group was not affected by the pre-treatment with bicuculline. From these findings, it is concluded that the involvement of DHA in stress responses may act via a GABA(A) receptor-mediated mechanism. PMID- 12573523 TI - The retinal anatomy and function of the myelin mutant taiep rat. AB - PURPOSE: To study the histology and the physiological function of the retina in the neurological myelin mutant, taiep rats during the postnatal developmental period (P20-P360). METHODS: Electroretinography (ERG) was applied to evaluate intensity dependence and spectral sensitivity of the responses to light. Retinal histology, morphometry, and immunocytochemistry were used to characterize the structure of the retina, with particular emphasis on the Muller (glial) cells. RESULTS: In the taiep rats of all ages studied, the scotopic ERG showed normal a- and b-wave amplitudes and latencies; likewise, the scotopic spectral sensitivity function was the same for control and taiep animals, with a maximal sensitivity (lambda(max)) at 500 nm. However, in adult taiep rats (P90 to P360) a secondary cornea-positive wave ('b(2)') was observed in response to high stimulus intensities, which never occurred in controls. This correlated with the observation that in the photopic ERG responses of the taiep rats, the b-wave was reduced in amplitude, and was followed by a rapid cornea-negative after potential. After 1 year of life, in taiep rats the outer plexiform layer (OPL) became slightly thinner and the inner plexiform/ganglion cell layers (IPL/GCL) appeared to be swollen, and increased in thickness; in addition, the number of retinal neurons (particularly, of photoreceptor cells) slightly decreased. Increased GFAP immunoreactivity revealed a hypertrophy and reactivity of the Muller cells in 1-year-old taiep rats. CONCLUSIONS: The present results suggest the occurrence of a relatively mild and slowly progressing neural retinal alteration in taiep rats, which becomes histologically and functionally evident at the end of the first year of life, and mainly affects the circuit(s) of the photopic ON-response. It is speculated that this alteration is due to missing/altered signals from demyelinated optic nerve. PMID- 12573524 TI - Glutaric acid induces oxidative stress in brain of young rats. AB - This study investigated the effects of glutaric acid, which predominantly accumulates in glutaric acidemia type I, on some in vitro parameters of oxidative stress in brain of young rats. We evaluated chemiluminescence, total radical antioxidant potential (TRAP) and the activities of the antioxidant enzymes catalase, glutathione peroxidase and superoxide dismutase in brain tissue homogenates in the presence of glutaric acid at concentrations ranging from 0.05 to 2.0 mM. The acid significantly increased chemiluminescence (up to 65%) and reduced total radical-antioxidant potential (up to 28%) and glutathione peroxidase activity (up to 46%), without affecting catalase and superoxide dismutase activities. The results provide evidence that glutaric acid induces oxidative stress in vitro in rat brain. If these findings also occur in humans, it is possible that they may contribute to the neuropathology of patients affected by glutaric acidemia type I. PMID- 12573525 TI - Cortico-striatal synaptic plasticity in endothelial nitric oxide synthase deficient mice. AB - Nitric oxide (NO) is a retrograde messenger involved in the processes of learning and memory. The role of the endothelial isoform of nitric oxide synthase (eNOS) in striatal synaptic plasticity was investigated in eNOS-deficient (eNOS(-/-)) and wild type (WT) mice. Tetanic stimulation of cortical afferents in WT mice evoked either long-term potentiation (LTP), or long-term depression (LTD) of cortico-striatal transmission. Both these plasticity related phenomena were NMDA receptor-dependent; LTD was blocked by sulpiride, a dopamine D2-receptor antagonist. LTP occurrence in slices from eNOS(-/-) mice was significantly reduced when compared with WT mice. The NOS inhibitor NL-ARG reduced the occurrence of LTP and increased the occurrence of LTD in WT mice, resembling the balance of LTP/LTD in eNOS(-/-) mice. Impairment of NO-synthesis thus shifts striatal plasticity towards LTD. This indicates a possible involvement of eNOS from endothelia in neuronal modulation. PMID- 12573526 TI - Globosa neurons: a distinct subgroup of noradrenergic neurons in the caudal pons of rats. AB - A previously undescribed subgroup of A7 neurons was identified and named globosa neurons. Morphologically, these neurons exhibit strong TH staining, are larger and globularly shaped, and are situated more laterally compared with the main group of A7 neurons. They have prominent dendritic processes that are oriented transversely and extend into the lateral lemniscus. These neurons are activated during underwater diving in rats, but at present their function is unknown. PMID- 12573527 TI - Mouse dexamethasone-induced RAS protein 1 gene is expressed in a circadian rhythmic manner in the suprachiasmatic nucleus. AB - We identified the Dexamethasone-induced RAS protein 1 (Dexras1) gene as a cycling gene in the suprachiasmatic nucleus (SCN). Investigation of the whole brain using in situ hybridization demonstrated the localization of the expression of the gene in the SCN, thalamus, piriform cortex and hippocampus. However, rhythmic expression of the gene was observed only in the SCN. The rhythmic change in gene expression during 1 day was approximately five-fold, and the maximum expression was observed during subjective night. Real-time PCR using the SCN, paraventricular nucleus and cortex confirmed these results. Next, we analyzed the expression of the Dexras1 gene in the SCN of cryptochrome (Cry) 1 and 2 double knockout mice. We found that the rhythmic expression disappeared. The results indicate that Dexras1 rhythmicity and levels are dependent upon CRYs. This is the first time that the G protein, which may be involved in the input pathway, has been isolated as a cycling gene in the SCN. PMID- 12573528 TI - Estrogen receptor beta (ERbeta) protein levels in neurons depend on estrogen receptor alpha (ERalpha) gene expression and on its ligand in a brain region specific manner. AB - Estrogen receptors (ERs) alpha and beta are very similar estrogen-binding proteins, perhaps gene duplication products, which act as ligand-dependent transcription factors. While the estrogenic regulation of ERalpha has been well documented, little is known about how estrogen regulates ERbeta and whether ERalpha plays a role in the expression and estrogenic regulation of ERbeta. In the present study, we examined the effects of gonadectomy and estrogen replacement on ERbeta immunoreactivity (ir) in wild-type (WT) and ERalpha knockout (alphaERKO) adult male mice in six brain regions, the medial preoptic area (MPOA), the bed nucleus of the stria terminalis (BNST), the paraventricular nucleus (PVN), the ventromedial nucleus of hypothalamus (VMH), the medial amygdala nucleus (MeAMY) and the dorsal raphe nucleus (DRN). Mice were divided into four different treatment groups: gonadally intact, gonadectomized (GDX), GDX+short-term treatment with estrogen (s.c. injection of estradiol benzoate (EB), 5 microg, at 48 h before perfusion) or GDX+long-term treatment with estrogen (implant of an EB pellet, 2.5 microg/day, for 10 days). In intact alphaERKO mice, the number of ERbeta expressing cells was significantly decreased in the MPOA and increased in the BNST, compared to WT mice. Both in the MPOA and BNST, steroid hormone regulation of ERbeta protein (an increase by GDX and a decline to intact levels by EB) was found only in WT, not in alphaERKO mice. In the VMH, GDX significantly increased the number of ERbeta ir expressing cells in both genotypes. EB treatment tended to decrease the number of ERbeta ir cells in WT mice, whereas EB treatment tended to increase ERbeta ir cell counts in alphaERKO mice. No effects of GDX or EB treatment were found in the DRN and MeAMY regardless of genotype. These results suggest that gonadal steroid hormones may regulate ERbeta protein in male mice and ERalpha may be involved in the expression and regulation of ERbeta in a region-specific manner. PMID- 12573529 TI - Exogenous myristoylated-G(i2)alpha subunits of GTP-binding proteins are mitogens following their internalization by astrocytes in culture. AB - Heterotrimeric GTP-binding proteins (G proteins) are involved in the coupling of a variety of cell surface receptors to different intracellular signalling pathways, some of which take part in the regulation of growth by affecting cell proliferation and/or differentiation. In cultured astrocytes, many receptors of neuropeptides and hormones are coupled to the heterotrimeric G(i) proteins which regulate the mitogen-activated protein kinase (MAPK/ERK) cascade through both the Galpha and Gbetagamma subunits. We have previously reported that functionally active recombinant myr-G(i2)alpha subunits added to such cultures are internalised and distributed within the plasma membrane and cytosol as well as in the nuclei of dividing astrocytes. Here we show that astrocytes proliferate dose dependently in response to exogenous myr-G(i2)alpha subunits. Concentrations of 100 pM-30 nM myr-G(i2)alpha caused more than 2.5-fold increase of [3H]thymidine incorporation over basal levels. Other classes of myr-Galpha subunits, such as G(i3)alpha or G(o)alpha, induced a much lower proliferative effect. The addition of G(i1)alpha subunits to the cultures produced no change, indicating the selectivity of this effect. Even though myr-G(i2)alpha subunits are internalised by the cells regardless of their guanine nucleotide-bound state, much less [3H]thymidine incorporation was observed in the presence of GDPbetaS-myr G(i2)alpha or GTPgammaS-myr-G(i2)alpha. Further, the fluorescent labelling was dissimilarly distributed, the signal being concentrated in the nucleus and perinuclear regions of the astrocytes. Selective disassembly of caveolae impaired both myr-G(i2)alpha internalisation and DNA induction. Together, these data reveal a proliferative effect of myr-G(i2)alpha subunits in astrocytes, and provide evidence for the incorporation of exogenous myr-G(i2)alpha subunits into the mitogen cascade activated by neurotransmitters or growth factors. The fact that Galpha proteins can enter cells is particularly interesting because options for delivering functional proteins into cells are limited. Thus, these proteins may have clinical applications for compensating deficits in the transduction mechanisms associated with several neurological diseases, or as a non-invasive membrane traversing carriers. PMID- 12573530 TI - Heteromer formation of delta2 glutamate receptors with AMPA or kainate receptors. AB - The delta2 glutamate receptor (GluRdelta2) is predominantly expressed in the postsynaptic densities of parallel fiber-Purkinje cell synapses and plays a crucial role in cerebellar function. However, the mechanisms by which GluRdelta2 participates in cerebellar functions are largely unknown because GluRdelta2 does not bind glutamate analogs. We investigated the possibility that GluRdelta2 may be involved in channel formation together with other glutamate receptor families. We transiently expressed lurcher mutant AMPA receptor GluR1(Lc) and kainate receptor GluR6(Lc) in HEK293 cells. Cells expressing these constitutively active channels displayed a rectifying current-voltage (I-V) relationship. However, when cells were co-transfected with GluRdelta2(Lc), which had the arginine residue in the channel pore region, cells displayed a linear I-V relationship, a result that indicates GluRdelta2(Lc) formed functional heteromeric channels with GluR1(Lc) or GluR6(Lc). Assembly of GluRdelta2 with GluR1 or GluR6 was further confirmed by co immunoprecipitation assays in HEK293 cells. In addition, GluRdelta2 receptors were partially co-immunoprecipitated from cerebellar synaptosomal fractions by antibodies against GluR2 or KA2. In contrast to lurcher channels, expression of wild-type GluRdelta2 significantly reduced the glutamate-induced current of the wild-type GluR1 receptors without affecting channel properties, such as current kinetics, dose-response relationship, and single-channel conductance. Thus, the heteromeric channel created by the association of wild-type GluR1 and GluRdelta2 may not be gated by glutamate and does not participate in glutamate-induced currents. These results suggest that GluRdelta2 and AMPA or kainate receptors can assemble to form heteromeric receptors in vitro and could modify glutamate signaling in vivo. These findings may help explain the role of GluRdelta2. PMID- 12573531 TI - Induction of mRNAs and proteins for Na/K ATPase alpha1 and beta1 subunits following hypoxia/reoxygenation in astrocytes. AB - Characteristics of the cellular response to oxygen deprivation and subsequent reoxygenation (hypoxia/reoxygenation) include redirection of energy metabolism, increased glucose utilization and expression of oxygen-regulated proteins. Inhibition of protein synthesis during early reoxygenation period prevented effective astrocyte adaptation to hypoxia/reoxygenation, resulting in eventual cell death. To elucidate the role of astrocytes in the central nervous system in response to hypoxia/reoxygenation, we analyzed the cDNA library derived from the cultured rat astrocytes subjected to 24 h of hypoxia followed by reoxygenation by differential display, and isolated a cDNA corresponding to Na/K ATPase alpha1 subunit. The expression of Na/K ATPase alpha1 subunit mRNA as well as beta1subunit mRNA was transiently increased after reoxygenation, whereas hypoxia itself did not induce any gene expression change. Na/K ATPase alpha1 subunit protein was transiently increased, whereas the protein expression for Na/K ATPase beta1 subunit showed sustained induction after reoxygenation. Overexpression of beta1 subunit in HEK 293 cells subjected to hypoxia/reoxygenation promoted survival of the cells. These findings suggest that Na/K ATPases may contribute to maintain the cellular environment of astrocytes subjected to hypoxia/reoxygenation. PMID- 12573532 TI - The Cu,Zn superoxide dismutase in neuroblastoma SK-N-BE cells is exported by a microvesicles dependent pathway. AB - The antioxidant enzyme Cu,Zn superoxide dismutase has so far been considered costitutively expressed and exclusively localized into cytosol. In this paper we investigated Cu,Zn superoxide dismutase export in neuroblastoma SK-N-BE cells by flow cytometry analysis, confocal immunofluorescence analysis and enzyme-linked immunosorbed assay. Immunofluorescence analysis shows that the enzyme is exported by microvesicular granules; moreover the treatment of cells with brefeldin A and with 2-deoxy-D-glucose and sodium azide strongly decreases the amount of CuZn superoxide dismutase detected in the medium. Therefore the involvement of ATP dependent mechanisms, likely including BFA-sensitive intracytoplasmic vesicles in Cu,Zn SOD export from SK-N-BE cells, has to be hypothesized. Microvesicular mediated Cu,Zn SOD export in neurons could represent a relevant phenomenon able to influence cell excitability that is affected by reactive oxygen species. PMID- 12573533 TI - Distribution and colocalization of NGF and GDNF family ligand receptor mRNAs in dorsal root and nodose ganglion neurons of adult rats. AB - To understand the dependence of primary sensory neurons on neurotrophic factors, we examined the distribution and colocalization of mRNAs for receptors of nerve growth factor (NGF) and glial cell line-derived neurotrophic factor (GDNF) family ligands in dorsal root ganglion (DRG) and nodose ganglion (NG) neurons of adult rats by in situ hybridization (ISH) histochemistry using serial sections. About 35, 10, and 20% of the lumbar DRG neurons expressed trkA, trkB and trkC mRNAs, respectively. Messenger RNA signals for c-ret, a common signaling receptor of GDNF family ligands, were seen in about 60% of DRG neurons, and some of these neurons expressed trkA, trkB, or trkC mRNAs. Most (97%) of the DRG neurons observed were positive to at least one of these four mRNAs. About 50, 20, and 20% of DRG neurons expressed GDNF family receptor alpha1 (GFR alpha1), GFR alpha2, and GFR alpha3 mRNAs, respectively, and most of these neurons were positive to c ret mRNA. Interestingly, GFR alpha2 and GFR alpha3 mRNA signals were frequently seen in the same neurons, which lack GFR alpha1 mRNA signals. On the other hand, 98% of NG neurons expressed trkB mRNA and 30-40% of NG neurons co-expressed c-ret and GFR alpha1 mRNAs. However, mRNA signals for other receptors (TrkA, TrkC, GFR alpha2, GFR alpha3) were seen in only a few NG neurons. These findings suggest that all the DRG neurons in adult rats depend on at least one of the NGF and GDNF family ligands, and that some DRG neurons depend on two ligands or more. In contrast, NG neurons were suggested to be divided into two major groups; one group depends on brain-derived neurotrophic factor (BDNF)/neurotrophin-4/5 (NT 4/5), and the other depends on both BDNF/NT-4/5 and GDNF. PMID- 12573534 TI - Hypothermia suppresses inducible nitric oxide synthase and stimulates cyclooxygenase-2 in lipopolysaccharide stimulated BV-2 cells. AB - Hypothermia is neuroprotective, possibly through suppression of microglial activation. We investigated the effects of hypothermia on lipopolysaccharide (LPS) stimulated BV-2 cells. At 37 degrees C, LPS elicited strong increases in inducible nitric oxide synthase (iNOS), nitric oxide (NO), cyclooxygenase-2 (COX 2), tumour necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6), accompanied by translocation of nuclear factor-kappaB (NF-kappaB) to the nucleus. Hypothermia (33 degrees C) caused complete suppression of iNOS and NO, a partial reduction of IL-6 but did not prevent TNF-alpha production or NF-kappaB translocation. In contrast, LPS induced cyclooxygenase-2 (COX-2) to higher levels under hypothermic conditions. These results show that hypothermia selectively suppresses iNOS in microglia. PMID- 12573535 TI - Oxidative stress regulated genes in nigral dopaminergic neuronal cells: correlation with the known pathology in Parkinson's disease. AB - Oxidative stress (OS) is a primary pathogenic mechanism of nigral dopaminergic (DA) cell death in Parkinson's disease (PD). Oxidative damage, Lewy body formation and decreased mitochondrial complex I activity are the consistent pathological findings in PD. In nigral DA neurons, however, it is unknown whether any gene expressional changes induced by OS contribute to the typical PD pathology. Here, using microarray analysis, we identified several groups of genes in the nigral DA cell line, SN4741 [J. Neurosci. 19 (1999) 10; J. Neurochem. 76 (2001) 1010], that were regulated by OS. Approximately 36 significantly regulated genes that encode functional molecules of nuclear subunits of mitochondrial complex I, exocytosis and membrane trafficking proteins, markers for OS and oxidoreductases, regulatory molecules of apoptosis and unidentified EST clones were further analysed. OS modulated the expression of specific genes, of which physiological dysfunctions have been implicated in PD. For instance, the expression of the nuclear-encoded subunits of mitochondrial complex I, B8 and B17, were significantly down-regulated by OS, possibly contributing to selective defect in mitochondrial complex I activity in PD. Furthermore, syntaxin 8 and heme oxygenase-1 (HO-1) are most dramatically up-regulated by OS in DA cells. Syntaxin 8 is a SNARE protein, regulating lipid vesicle docking and fusion as well as early endosome membrane recycling. Lipid membranes are significantly oxidative-damaged in PD. HO-1 is an important cytoplasmic constituent of Lewy bodies, a pathological hallmark of idiopathic PD. Thus, our findings provide novel molecular probes that may be useful in unraveling the molecular mechanism(s) of OS-induced pathogenesis in PD. Further functional characterization of the affected genes including ESTs can help elucidate the underlying molecular pathology as well as develop biomarkers for monitoring degenerating DA neurons in PD. PMID- 12573536 TI - Methylmercury-induced reactive oxygen species formation in neonatal cerebral astrocytic cultures is attenuated by antioxidants. AB - Excessive generation of reactive oxygen species (ROS) has been suggested as a causal factor in various neurodegenerative disorders, such as Parkinson's disease and Alzheimer's disease [Brain Res. 830 (1999) 10-15; Biochem. J. 310 (1995) 83 90; Free Radic. Biol. Med. 27 (1999) 612-616]. The present work examined the role of ROS in the neurotoxicity of methylmercury (MeHg). ROS formation in primary astrocytic cultures of neonatal rat cerebral cortex was monitored by 2',7' dichlorodihydrofluorescein diacetate (H(2)DCF-DA) fluorescence. MeHg, at 10 and 20 microM caused a significant increase in ROS formation (10 microM, P<0.01; 20 microM, P<0.001). Additional studies established the effectiveness of antioxidants/free radical scavengers in attenuating the MeHg-stimulated ROS formation in the following rank-order: (1) Trolox (6-hydroxy-2,5,7,8 tetramethylchroman-2-carboxylic acid), a non-thiol containing antioxidant, (2) n propyl gallate (PG), a free radical scavenger, (3) superoxide dismutase (SOD), an antioxidant enzyme that dismutates superoxide anion radical, (4) alpha-phenyl tert-butyl nitrone (PBN), a lipophilic hydroxyl radical spin trapping agent. A significant inhibition of MeHg-induced ROS generation was also noted in astrocytes preincubated (3 h) with arachidonyl trifluoromethyl ketone (AACOCF(3,) 20 microM, P<0.05), a specific inhibitor of cytosolic phospholipase A(2) (cPLA(2)). Conversely, pretreatment (24 h) with 100 microM buthionine-L sulfoxamine [BSO, a glutathione (GSH) synthesis inhibitor], significantly increased (P<0.05) ROS formation in MeHg treated astrocytes compared to controls. Combined, these studies invoke ROS as potent mediators of MeHg cytotoxicity and support the hypothesis that excessive ROS generation, at least in part, plays an important role in MeHg-induced neurotoxicity. PMID- 12573537 TI - Activation of microglia with lipopolysaccharide leads to the prolonged decrease of conventional protein kinase C activity. AB - The activation of microglia by lipopolysaccharide (LPS) results in the induction of harmful factors including nitric oxide and tumor necrosis factor alpha (TNFalpha). Such microglial activation was suggested to be mediated by PKC activity based on the results of an inhibitor experiment. To clarify the relationship between microglial activation and PKC activity, conventional PKC (cPKC) activity was measured by enzyme-linked immunosorbent assay (ELISA) in LPS activated microglia. LPS stimulation caused a time- and dose-dependent decrease (70%) of specific activity of cPKC, ascribed to the decreasing amounts of PKCalpha. However, the remaining PKC activity (30%) was sustained despite longer incubation or higher LPS concentration. Therefore, it is suggested that LPS stimulated microglia require priming by PKC activation for the induction of harmful factors, while only a part (30%) of original PKC activity is sufficient for durable microglial activation. PMID- 12573538 TI - Dopamine transporters are dephosphorylated in striatal homogenates and in vitro by protein phosphatase 1. AB - Dopamine transporters (DATs) undergo increased phosphorylation upon treatment of striatal tissue or cultured cells with protein kinase C activators and protein phosphatase inhibitors. Phosphorylation conditions also lead to reductions in dopamine transport activity, which may function to regulate synaptic dopamine levels and control the extent and duration of dopaminergic signaling. Treatment of rat striatal tissue with okadaic acid (OA), a broad-spectrum protein phosphatase inhibitor, produces apparent maximal increases in DAT phosphorylation, suggesting that dephosphorylation is a crucial regulator of the DAT phosphorylation state. We used a combination of endogenous and in vitro approaches to identify the phosphatase(s) responsible for this activity. In homogenates prepared from (32)PO(4)-labeled rat striatal slices, OA inhibited DAT dephosphorylation with an IC(50) of 40 nM, a dose most compatible with inhibition of protein phosphatase 1 (PP1). Dephosphorylation of DAT in striatal homogenates was also inhibited by PP1 inhibitor 2, while little effect was produced by protein phosphatase 2A inhibitor 1. In vitro dephosphorylation assays showed substantial removal of (32)PO(4) from DATs by PP1 but not by protein phosphatase 2A, protein phosphatase 2B, or protein tyrosine phosphatase, and this effect was blocked by OA, verifying that the (32)PO(4) loss from DAT was due to dephosphorylation. These results demonstrate that DAT is a direct substrate for PP1 in vitro and suggest that PP1 is a major DAT phosphatase in rat striatum. PMID- 12573539 TI - Morphine withdrawal is modified in pituitary adenylate cyclase-activating polypeptide type I-receptor-deficient mice. AB - The pituitary adenylate cyclase-activating polypeptide type I-receptor (PAC1) is a G-protein-coupled receptor that is widely expressed in neurons of the central and peripheral nervous system. The strong expression of PAC1 in the second sensory neuron as well as in brainstem regions such as the locus coeruleus prompted us to elucidate the potential in vivo role of PAC1-mediated signalling in pain perception and opioid addiction using a PAC1-deficient mouse line. We observed a selective involvement of PAC1 in the mediation of visceral pain. While there was no impairment in acute somatic pain perception, PAC1-mutants exhibited a dramatically decreased response in the abdominal writhing test. These data in concert with data from the literature implicate PAC1 in the mediation of visceral and chronic pain. In addition, we observed that PAC1 did not influence the motivational aspects of opioid addictive properties, since morphine-induced rewarding effects and sensitization to locomotor responses were completely maintained in PAC1-deficient mice. However, there was a dramatic increase in physical withdrawal signs after naloxone-precipitated morphine withdrawal in PAC1 mutants. At the cellular level, electrophysiological examinations in locus coeruleus neurons from morphine-dependent wild-type and PAC1-deficient mice did not reveal any differences in firing rates. These data therefore suggested that most likely disruption of PAC1-mediated signalling in afferents towards the locus coeruleus but not within the intrinsic locus coeruleus system led to the enhancement of somatic withdrawal signs. PMID- 12573540 TI - Co-localization of cholesterol, apolipoprotein E and fibrillar Abeta in amyloid plaques. AB - Recent evidence strongly suggests a role for cholesterol and apolipoprotein E in the etiology of Alzheimer's disease. We have demonstrated the co-localization of cholesterol and apolipoprotein E with beta-amyloid immunoreactivity and thioflavin S immunofluorescence in AD type plaques of a transgenic mouse model. Cholesterol and apolipoprotein E co-localized to the core of thioflavin S positive (fibrillar) plaques, but not thioflavin S-negative (diffuse) plaques from an early age. By 18 months of age, there was extensive coverage of fibrillar plaques immunopositive for apolipoprotein E and cholesterol oxidase. These findings support evidence that cholesterol and apolipoprotein E are involved in fibrillar plaque formation or maintenance, and suggest that cholesterol may impact amyloid formation extracellularly, as well as through an intracellular effect. PMID- 12573541 TI - Mouse vesicular GABA transporter gene: genomic organization, transcriptional regulation and chromosomal localization. AB - The vesicular GABA transporter (VGAT) loads GABA from neuronal cytoplasm into synaptic vesicles and is selectively expressed in inhibitory neurons that contain GABA and/or glycine. To elucidate the molecular mechanisms of mouse VGAT (mVGAT) gene expression, we have isolated and characterized the mVGAT gene. The mVGAT gene was found to be 4.7 kilobases in size and to contain three exons and two introns by comparison of the cloned genomic DNA with the cDNA (termed mVGATa) sequence reported by Sagne et al. [FEBS Lett. 417 (1997) 177]. Analysis of transcripts and genomic DNA revealed an alternatively spliced mVGAT isoform (termed mVGATb) that retains intron 2 of mVGATa as an exon. This alternative transcript specifies 514 amino acid residues identical to VGATa followed by a unique C-terminal sequence of 11 amino acids encoded by intron 2. Fluorescent in situ hybridization studies showed that the mVGAT gene is localized on chromosome 2. One major transcription start site of the mVGAT gene is an A residue 209 bp upstream from the translational initiation site, as shown using the 5'-RACE method. RT-PCR analysis revealed that the mVGAT gene was expressed at a high level in retinoic acid-treated P19 embryonal carcinoma cells, at a very low level in non-treated P19 cells, and not detectably expressed in Neuro-2a neuroblastoma cells. Sequence analysis of the 5'-flanking region revealed a number of putative regulatory elements including Sp1, Egr-1 and Pitx binding sites. In transient transfection assays, 2 kilobases of the mVGAT 5'-flanking region generated similar levels of luciferase reporter activity in three kinds of cultured cells. Deletion analysis and gel mobility shift assays demonstrated that the region -161 to +155 contained the basal promoter activity of the mVGAT gene and that an activating region from -49 to -27 bound an Sp1-like protein. These results suggest a possible mechanism for regulation of the expression of the mVGAT gene. PMID- 12573542 TI - Chromatin condensation during glutamate-induced excitotoxicity of cerebellar [correction of celebellar] granule neurons precedes disintegration of nuclear DNA into high molecular weight DNA fragments. AB - The disturbance of the intracellular ionic homeostasis after activation of channel-associated membrane receptors by the excitatory neurotransmitters represents a principle event that triggers excitotoxic cell death of neurons. Here we demonstrate that glutamate-induced excitotoxicity of cerebellar granule neurons was accompanied by apoptosis-like nuclear shrinkage, chromatin condensation, and disintegration of nuclear DNA into high molecular weight DNA fragments, but was neither associated with activation of caspase 1, -2, -3, -9, nor was protected by a pan-caspase inhibitor, zVAD-fmk. We further demonstrate that chromatin condensation took place at the early stages of excitotoxicity and preceded nuclear DNA fragmentation. The results suggest that fragmentation of nuclear DNA and condensation of chromatin are uncoupled events during neuronal cell death PMID- 12573543 TI - Prolonged expression of c-Fos and c-Jun in the cerebral cortex of rats after deltamethrin treatment. AB - In this study we investigated the effects of deltamethrin on the expression of c Fos and c-Jun in the cerebral cortex of rats. Immunohistochemical analysis demonstrated that the immunoreactivity for c-Fos was markedly increased in the cerebral cortex 5 h after deltamethrin treatment, and maintained at an increased level at 24 h, even though little immunoreactivity for c-Fos was seen in the same brain region of control rats. The immunostaining for c-Jun was also dramatically elevated in the same brain region, showing the same time course of c-Fos expression after deltamethrin treatment. Further, both MK-801, an N-methyl-D aspartate (NMDA) receptor antagonist, and NBQX, an alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionate (AMPA)/kainate (KA) receptor antagonist, attenuated deltamethrin-elicited prolonged expression of c-Fos and c-Jun. Since the persistent expression of c-Fos and c-Jun is unusual, and has been reported before in conditions involving neurodegeneration, our results are consistent with a model that deltamethrin induces neurodegeneration through a glutamate-dependent pathway. PMID- 12573544 TI - Mice expressing human mutant presenilin-1 exhibit decreased activation of NF kappaB p50 in hippocampal neurons after injury. AB - Mutations in the presenilin-1 (mutPS-1) gene, a cause of familial Alzheimer's disease, increase the susceptibility of neurons to apoptotic death. Using the trimethyltin model of hippocampal neurodegeneration, mice expressing the human mutPS-1 gene (M146L) exhibited increased neurodegeneration and mortality relative to non-transgenic littermates. Activation of NF-kappaB p50 was found to be impaired in transgenic mice with unaltered expression levels suggesting that mutPS-1 expression inhibits p50 activation to adversely affect neuronal resistance to injury. PMID- 12573545 TI - Moellerella wisconsensis: identification, natural antibiotic susceptibility and its dependency on the medium applied. AB - The present study establishes a data compilation on biochemical features and natural antibiotic susceptibilities of Moellerella wisconsensis strains. 17 moellerellae isolated from humans (n = 11), food (n = 5) and water (n = 1) were tested. Identification was carried out using two commercially available systems and conventional tests. MIC determinations of 74 antibiotics were performed applying a microdilution procedure in Cation-adjusted Mueller Hinton broth and IsoSensitest broth. M. wisconsensis was naturally sensitive to doxycycline, minocycline, all tested aminoglycosides, numerous beta-lactams, all fluoroquinolones, folate-pathway inhibitors, chloramphenicol and nitrofurantoin. Natural resistance was found with oxacillin, penicillin G, all tested macrolides, lincomycin, streptogramins, ketolides, glycopeptides, fusidic acid, linezolid and rifampicin. Medium-dependent differences in susceptibility affecting clinical assessment criteria were seen with tetracycline, clindamycin and fosfomycin. From the data of the present study it is possible that some moellerellae are misidentified as Klebsiella pneumoniae subsp. ozaenae. PMID- 12573546 TI - Isolation of methicillin-resistant coagulase-negative staphylococci from patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and comparison of different molecular techniques for discriminating isolates of Staphylococcus epidermidis. AB - Coagulase-negative staphylococci (CNS) have emerged as an important pathogen in nosocomial infections. About 80%-90% of CNS isolates associated with hospital infections are methicillin-resistant coagulase-negative staphylococci (MRCNS). The aims of this study were to screen for MRCNS isolates in the flora of a small population of patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and to evaluate the discriminatory power of different molecular methods: pulsed-field gel electrophoresis (PFGE), mecA location, ClaI/mecA polymorphism and arbitrarily primed polymerase chain reaction (AP-PCR) for characterizing isolates of methicillin-resistant Staphylococcus epidermidis (MRSE). Seventy-nine CNS isolates were recovered from the 11 CAPD patients studied. Using a methicillin screening agar and a DNA specific mecA probe we verified that 30 of the 79 (38%) CNS isolates were resistant to methicillin (MRCNS). Twenty-two of the 30 MRCNS (73%) were MRSE, 7 (23%) methicillin-resistant S. haemolyticus (MRSH(ae)) and 1 (3%) methicillin-resistant S. hominis (MRSH(om)). All patients analyzed carried MRCNS in their flora, in one or more sites. Since CAPD patients have high risk for developing peritonitis, the colonization of these patients with MRCNS might represent an additional problem, due to the therapeutic restrictions imposed by these multiresistant isolates. A wide genetic diversity was verified when the PFGE of the MRSE isolates was analyzed. The 22 MRSE isolates displayed a total of 15 PFGE different patterns (11 PFGE types and 4 subtypes). The location of mecA in the SmaI-fragmented genome DNA did not bring any additional advantage for epidemiologic characterization of the isolates. The ClaI/mecA polymorphism was able to correctly discriminate 12 from the 15 PFGE patterns. In addition, the DNA of 20 MRSE isolates were used for AP-PCR typing. These isolates belonged to 14 PFGE patterns (11 types and 3 subtypes) and displayed 15 genotypes (for the association of PFGE, mecA location and ClaI/mecA polymorphism). A total of 17 different amplification patterns was verified using the primer 1. Only for 2 genotypes, strains having identical genetic backgrounds were further discriminated by AP-PCR (2 of 15 genotypes (87%) for AP-PCR and 1 of 15 genotypes for PFGE; (93%). Concluding, our results indicated that the AP-PCR can be an alternative and useful tool for monitoring and genotyping MRSE colonization and also to molecular characterizing MRSE outbreaks in hospitals. PMID- 12573547 TI - Detection of group B streptococcal bacteremia in simulated intrapartum antimicrobial prophylaxis. AB - The diagnostic value of negative blood cultures from neonates whose mothers receive intrapartum antimicrobial prophylaxis for prevention of perinatal group B streptococcal disease is uncertain. We investigated whether blood culture medium containing resin designed to adsorb antibiotic improved group B streptococcal detection following simulated intrapartum antimicrobial prophylaxis. Group B streptococcus (Streptococcus agalactiae) was preincubated with varying antibiotic concentrations before inoculation into BACTEC Peds Plus resin-containing medium, BACTEC Standard, or Trek ESP 80A. In the presence of 10 mcg/mL ampicillin, detection of both low (<500 CFU/mL) and high (>500 CFU/mL) S. agalactiae inocula ranged between 75-100% of resin-containing medium bottles; detection rates in both non-resin-containing media were lower. When S. agalactiae was detected, it was detected sooner with resin-containing medium. The addition of gentamicin to ampicillin did not affect sensitivity of resin-containing medium for S. agalactiae. In our model, resin-containing medium more consistently and more rapidly detected S. agalactiae than did either of two non-resin-containing media, in the presence of antibiotic levels likely present in fetal sera following intrapartum antimicrobial prophylaxis. PMID- 12573548 TI - Comparison of BDPhoenix and VITEK2 automated antimicrobial susceptibility test systems for extended-spectrum beta-lactamase detection in Escherichia coli and Klebsiella species clinical isolates. AB - The present study compares the ability to detect extended-spectrum beta lactamases (ESBL) among a collection of 34 ESBL producing clinical isolates belonging to Escherichia coli and Klebsiella species with two new rapid susceptibility and identification instruments-VITEK2 (bioMerieux, Marcy l'Etoile, France) vs. BDPhoenix (BD Biosciences, Sparks, MD). ESBL content in these isolates was previously characterized on the basis of PCR amplification and sequencing results which were used as the reference method in our evaluation. BDPhoenix correctly determined the ESBL outcome for all strains tested (100% detection rate), whereas VITEK2 was not able to detect the ESBL status in 5 isolates (85% detection rate). Detailed analysis revealed that the discrepancies were mainly observed with 'difficult-to-detect' strains. Misidentification was either due to low oximino cephalosporin MIC in these strains or was associated with pronounced 'cefotaximase' or 'ceftazidimase' phenotypes. Klebsiella oxytoca chromosomal beta-lactamase (K1) is phenotypically quite similar to ESBL enzymes. In order to evaluate whether the K1 and ESBL enzymes could be discriminated, we expanded our analysis by 8 clinical K. oxytoca strains with K1 phenotypes. VITEK2 gave excellent identification of these strains whereas 7 out of 8 were falsely labeled ESBL-positive by the BDPhoenix system. PMID- 12573549 TI - Detection and characterization of leishmania antigens from an American cutaneous leishmaniasis vaccine for diagnosis of visceral leishmaniasis. AB - Antigens were isolated from vaccines against American Cutaneous Leishmaniasis (ACL) and their reactivity tested against nine different groups of human sera and two groups of dog sera. These antigens react specifically with human and dog visceral leishmaniasis sera when compared to sera from non-infected individuals. Sera from humans from endemic areas of ACL before, or one year after, vaccination, and ACL patients treated and cured by immunotherapy with polyvalent vaccine, did not display significant differences of reactivity to these antigens. In contrast, they displayed a significantly higher reactivity to the antigens when compared to sera from healthy humans from non-endemic areas. No sera reactivity was observed with patients carrying Chagas' disease or tuberculosis. These antigens are polysaccharides aggregates and present molecular masses ranging from 90 to over 200 KDa. These data suggest the use of these antigens for sero-diagnosis of human and canine visceral leishmaniasis. PMID- 12573550 TI - Igm recognition of recombinant Toxoplasma gondii antigens by sera of acutely or latently infected humans. AB - Clinical non-relevant (CNR) IgM specific for Toxoplasma gondii is responsible for false-positive results in commercially available IgM assays. Using IgM immunoblotting, it is possible to distinguish between IgM in sera of acutely infected (AI) patients and CNR IgM. Especially the combination of staining of a 55 and 30 kD antigen in T.gondii lysate proved useful in this respect. The 55 kD antigen was identified as Rop1, while the 30 kD antigen was confirmed to be Sag1. However, the use of recombinant antigens instead of lysates for diagnostic assays would improve reproducibility. IgM recognized recombinant Rop1, but most CNR sera also had low anti-Rop1 titers. Although purified native Sag1 separated AI and CNR sera very well on immunoblot, IgM did not recognize recombinant Sag1 at all. Clearly, it is difficult to produce a recombinant Sag1 that can be recognized by IgM. Recombinant Rop1 might be suitable as one of the recombinant antigens in an IgM immunoblot assay, but has to be combined with at least one other immunogenic antigen. PMID- 12573551 TI - Detection and drug-susceptibility testing of M. tuberculosis from sputum samples using luciferase reporter phage: comparison with the Mycobacteria Growth Indicator Tube (MGIT) system. AB - Rapid diagnosis of drug-resistant M.tuberculosis (Mtb) is desirable worldwide. We (i) describe a new luciferase reporter phage (LRP), phAE142 for this purpose; (ii) compare it to the automated MGIT 960 for time-to-detection of Mtb in clinical specimens; and (iii) evaluate its use for species confirmation and antibiotic susceptibility testing(AST) of Mtb. Twenty sputum samples were inoculated for testing by LRP, or by MGIT 960. After "positives" were identified by either method, the LRP was used for confirmation of Mtb complex (TBC) and for AST. The LRP method proved comparably efficient to MGIT 960 at detecting Mtb. Using an antibiotic uniquely inhibiting TBC with LRP provided species assignment, concurrently with AST, in a median of 3 days, with a sensitivity of 97%. Overall agreement in susceptibility results was 96%. Reliable susceptibility results and identification of TBC can be completed in a median of 12 days (range 8 to 16d) with LRP applied to sputum samples. PMID- 12573553 TI - Antimicrobial susceptibility testing of Campylobacter jejuni: a comparison between Etest and agar dilution method. AB - The susceptibility of Campylobacter jejuni strains (n = 50) against nine antimicrobials were determined in comparison with Etest (AB BIODISK, Solna, Sweden) and agar dilution method to further investigate the correlation between the two methods. All the strains were isolated from stool samples of patients with diarrhea in 1998 and found to be highly susceptible (>84%) to ampicillin, tetracycline, gentamicin, chloramphenicol, ciprofloxacin and erythromycin. The essential agreement between two methods was 66.6% (+/-1 log(2) dilution) and 85.5% (+/-2 log(2) dilution). The agreement of susceptibility categories was higher at 94.4%. PMID- 12573552 TI - Molecular characterization of fluoroquinolone resistant Streptococcus pneumoniae clinical isolates obtained from across Canada. AB - There is little published data detailing fluoroquinolone resistance in clinical isolates of S. pneumoniae. The purpose of this study was to characterize the resistance mechanisms of 34 fluoroquinolone-resistant S. pneumoniae clinical isolates obtained from medical centers in 8 of 10 Canadian provinces between 1997 and 2000. The quinolone resistance determining regions of gyrA, parC, and parE from the isolates were sequenced. The isolates were evaluated for reserpine sensitive efflux of ciprofloxacin and the new fluoroquinolones: gatifloxacin, gemifloxacin, levofloxacin and moxifloxacin. The isolates were typed using pulsed field gel electrophoresis. The majority of the isolates were genetically unrelated. Lower level fluoroquinolone resistance (ciprofloxacin MIC 4-8 microg/ml) was associated with amino acid substitutions in ParC, while higher level resistance (ciprofloxacin MIC > or = 16 microg/ml) was associated with amino acid substitutions in both ParC and GyrA. ParE substitutions were not associated with clinical resistance. Twelve of 34 (35%) isolates demonstrated reserpine-sensitive efflux of ciprofloxacin. Efflux alone conferred low level ciprofloxacin resistance in 3 isolates. Significant reserpine-sensitive efflux of the new fluoroquinolones was not observed. PMID- 12573554 TI - Determination of disk diffusion and MIC quality control parameters for AZD2563, a novel long-acting oxazolidinone. AB - AZD2563 is a new, long-acting oxazolidinone that possesses potent antimicrobial activity against Gram-positive organisms. This report summarizes the results of broth microdilution MIC and disk diffusion quality control (QC) investigations for AZD2563. All tests or methods (M2-A7 and M7-A5) and QC study designs (M23-A2) were those published by the National Committee for Clinical Laboratory Standards. QC strains tested against AZD2563 included: Staphylococcus aureus ATCC 25923 and ATCC 29213, Enterococcus faecalis ATCC 29212 and Streptococcus pneumoniae ATCC 49619. In the eight-center QC trial, 320 and 480 results were generated for MIC and disk diffusion tests, respectively. The selected 7-mm ranges for the two disk diffusion QC organisms contained > or = 98.5% of reported results. Proposed 3 or 4 log(2) dilution MIC ranges contained all reported participant results. AZD2563 disk diffusion zone diameters were generally 2-3 mm smaller than the corresponding linezolid zone diameters. In conclusion, QC ranges for AZD2563 MIC and disk diffusion methods were established and appear acceptable in preparation for the clinical trials use. PMID- 12573555 TI - Emergence of an IMP-like metallo-enzyme in an Acinetobacter baumannii clinical strain from a Brazilian teaching hospital. AB - Multidrug-resistant Acinetobacter spp. constitute a serious cause of nosocomial infection in Brazilian hospitals. This manuscript reports the first appearance of an IMP-like metallo-beta-lactamase encountered in a clinical isolate of A. baumannii from a Brazilian teaching hospital. PMID- 12573556 TI - Real-time PCR for the rapid detection of vanA and vanB genes. AB - A real-time PCR assay suitable for use on the Roche LightCycler platform was developed to replace an existing gel-based PCR assay for the simultaneous detection of the vanA & vanB genes in enterococcal isolates. Novel Fluorescence Resonance Energy Transfer (FRET) hybridization probes were designed. The multiplex real-time PCR assay and the existing gel-based assay were 100% concordant and both correctly detected the vanA or vanB genes in 4/4 VanA E. faecium and 25/25 VanB E. faecium. Additionally, 1/1 VanC1 E. gallinarum, 1/1 VanC2 E. casseliflavus and 47/47 vancomycin susceptible enterococci were negative for the vanA and vanB genes in both PCR assays. Results were available within 1.5 h for the real-time PCR assay compared to up to 5.5 h for the conventional PCR assay. PMID- 12573557 TI - Separation of cashew (Anacardium occidentale L.) nut shell liquid with supercritical carbon dioxide. AB - Cashew nut shell liquid (CNSL) represents the largest readily available bioresource of alkenyl phenolic compounds. In this work, separation of CNSL from the pericarp of the cashew nut with supercritical carbon dioxide was studied. In the initial extractions with CO(2) at 40-60 degrees C and at pressures from 14.7 to 29.4 MPa, low yields were obtained. However, when the extractions were performed with one or more intermediate depressurization steps, the yield of CNSL increased to as high as 94%. Most of the oil did not separate from the shell during the depressurization step, but was obtained during the subsequent repressurization. The CNSL extract had a clear light brownish pink color and exhibited no evidence of polymerization or degradation. The pressure profile extraction method proposed in this work increases the possible CNSL extraction yields and greatly reduces the amount of CO(2) required for CNSL separation. PMID- 12573558 TI - Xylan-hydrolyzing enzyme system from Bacillus pumilus CBMAI 0008 and its effects on Eucalyptus grandis kraft pulp for pulp bleaching improvement. AB - The extracellular productions of beta-xylanase, beta-xylosidase, beta glucosidase, beta-mannanase, arabinosidase, alpha-glucuronidase, alpha galactosidase and Fpase from Bacillus pumilus CBMAI 0008 were investigated with three different xylan sources as substrate. The enzymatic profiles on birchwood, Eucalyptus grandis and oat were studied at alkaline and acidic pH conditions. B. pumilus CBMAI 0008 grown on the three carbon sources produced mainly beta xylanase. At pH 10, the levels of xylanase were 328, 160 and 136 U/ml, for birch, oat and E. grandis, respectively. beta-Mannanase production was induced on E. grandis (5 U/ml) and arabinofuranosidase on oat (5 U/ml). Although small quantities of alpha-glucuronidase had been produced at pH 10, activity at pH 4.8 was 1.5 U/ml, higher than observed for Aspergillus sp. in literature reports. Preliminary assays carried out on E. grandis kraft pulp from an industrial paper mill (RIPASA S.A. Celulose e Papel, Limeira, SP, Brazil) showed a reduction of 0.3% of chlorine use in the pulp treated with the enzymes, resulting in increased brightness, compared to conventional bleaching. The enzymes were more efficient if applied before the initial bleaching sequence, in a non-pre-oxygenated pulp. PMID- 12573559 TI - Availability of corn stover as a sustainable feedstock for bioethanol production. AB - The amount of corn stover that can be sustainably collected is estimated to be 80 100 million dry tonnes/yr (t/yr), a majority of which would be available to ethanol plants in the near term as only a small portion is currently used for other applications. Potential long-term demand for corn stover by non fermentative applications in the United States is estimated to be about 20 million dry t/yr, assuming that corn stover-based products replace 50% of both hardwood pulp and wood-based particleboard, and that 50% of all furfural production is from corncobs. Hence, 60-80 million dry t/yr of corn stover should be available to fermentative routes. To achieve an ethanol production potential of 11 billion L (3 billion gal) per year (a target level for a non-niche feedstock), about 40% of the harvestable corn stover is needed. This amount should be available as long as the diversion of corn stover to non-ethanol fermentative products remains limited. PMID- 12573560 TI - Unconverted chars obtained during biomass gasification on a pilot-scale gasifier as a source of activated carbon production. AB - Biomass gasification was used to produce activated carbon on a pilot-scale fluidised-bed gasifier. The feedstock included both biomass alone and biomass mixed with coal and coal/granulated plastic wastes. This paper reports the results obtained from four different runs undertaken under various conditions of fuel supply, different ratios of steam/air for the gasification and temperature. These conditions were selected because they led to a significant amount of unconverted chars produced during gasification (from 0.72 to 1.4 kg) which then served as raw material for the production of activated carbon whilst the amount of gas obtained was also high enough for its potential use for different end-use applications. From the analysis of the results obtained, it can be concluded that a reasonable porosity development (mainly in the area of narrow micropores) was obtained by gasifying unblended pine wastes with steam for 4 h, producing about 1.4 kg of good-quality activated carbon (micropore volume of 0.263 cm(3)/g). In other runs, chars with a reduced microporosity development (i.e. 0.180 cm(3)/g) were obtained, however, they could be used as a proper starting material for the chemically activated carbon production. PMID- 12573561 TI - Bamboo as bioresource in Ethiopia: management strategy to improve seedling performance (Oxytenanthera abyssinica). AB - Seedling emergence and subsequent survival and growth are vital for natural forest restoration or plantation establishment by means of seeds. Such information is lacking for the African bamboo species. Two experiments were carried out in a greenhouse to evaluate the influence of seed orientation and sowing depth of the lowland bamboo Oxytenanthera abyssinica on seedling emergence, survival and growth. A randomised complete block design was used. Seedling emergence in the seed orientation experiment followed the order embryo end-up>lay-flat>embryo-end-down. Survival rate after 62 days decreased in the order lay-flat>embryo-end-down>embryo-end-up. Mean seedling height and number of leaves per seedling followed a similar pattern. Seeds sown on top of the soil surface and at 2.5 mm depth achieved faster and higher seedling emergence than those sown at 5 and 10 mm depths. However, mean seedling height and number of leaves per seedling were higher in 5 and 2.5 mm depths than surface and 10 mm depths. There were significant quadratic relationships between sowing depth and seedling height (p=0.034) as well as number of leaves per seedling (p=0.032), both peaking around 5 mm soil depth. Lay-flat orientation, which is the most frequent position in broadcast sowing, is recommended at 5 mm sowing depth for the lowland bamboo based on overall performance in seedling emergence, survival and growth. PMID- 12573562 TI - Determination of organochlorine pesticides in agricultural soil with special reference to gamma-HCH degradation by Pseudomonas strains. AB - Soil samples were taken from different agricultural fields and analyzed for organochlorine pesticide residues by gas chromatography. The analysis indicated that the soil samples contained some common organochlorine pesticides DDT, DDD, DDE, HCH and Aldrin. gamma-HCH was detected as 47.35 ppb whereas the concentrations of alpha-HCH, beta-HCH, p('),p(')-DDE, o('),p(')-DDT were 38.81, 1.79, 7.10 and 13.30 ppb, respectively, in the same soil. Two Pseudomonas strains isolated from agricultural soil were found to possess gamma-hexachlorocyclohexane degrading ability when the isolates were grown in a mineral salt medium containing gamma-HCH as the sole source of carbon and a number of metabolites were produced and detected by the gas chromatography. These bacterial isolates were further tested for carbohydrate and amino acid utilization as well as for their susceptibility against 10 commonly used antibiotics namely amoxycillin, chloramphenicol, cloxacillin, doxycycline, methicillin, nalidixic acid, neomycin, nitrofurantoin, streptomycin and tetracycline. Both the isolates were also screened for plasmid DNA and found to harbour a single plasmid. PMID- 12573563 TI - Thermal composting of faecal matter as treatment and possible disinfection method -laboratory-scale and pilot-scale studies. AB - When using toilets where the urine and faeces are collected separately for reuse as nutrients in agriculture, the collected matter should be disinfected. One way to do this is by thermal composting. Composting of different material mixes was investigated in a laboratory-scale experiment. This showed that the best mixture for dry thermal composting was a mix of faeces, food waste and amendment. The urine was collected separately by use of urine-diverting toilets. A new method was developed to mathematically evaluate and estimate the safety margins of pathogen inactivation during thermal composting. The method is based upon a mathematical calculation of the number of times total inactivation (at least 12log(10) reduction) of the organisms is achieved. In a pilot-scale experiment, the disinfection of a faeces/food waste mix was performed with a calculated safety margin of more than 37 times the total die-off of Enteroviruses and some 550 times that of Ascaris. Thus, well functioning composting seems to be effective for disinfection of faecal matter. To get a high temperature in all of the material, the reactor has to have sufficient insulation. A major disadvantage is the initial need for handling the raw un-disinfected material. The degradation of the organic matter in the compost was almost 75%, resulting in a small final volume that could safely be recycled. PMID- 12573564 TI - Preparation of spent brewer's yeast beta-glucans with a potential application as an immunostimulant for black tiger shrimp, Penaeus monodon. AB - Spent brewer's yeast was autolysed and the insoluble yeast cell wall fraction obtained was used as a raw material for the preparation of brewer's yeast beta glucan (BYG). A simple alkaline extraction was applied and optimized. The BYG with significantly (P 0.05) in progeny viruses grown in TK-expressing cells and control cells. Therefore, both LacZ and SupF mutagenesis assays demonstrated that TK of the HSV-1 strain KOS did not have detectable mutator activity. PMID- 12573585 TI - T helper cell-independent antibody responses to the transgene product of an e1 deleted adenoviral vaccine require NK1.1 T cells. AB - Mice lacking CD4(+) T cells due to a knock-out mutation respond to vaccination with a replication-defective adenoviral recombinant expressing the glycoprotein of rabies virus with a long-lasting virus-neutralizing antibody response. The vaccine-induced B cells secrete antibodies that are mainly of IgG isotypes. The response can be enhanced upon booster immunization, indicating the induction of B cell memory in the absence of CD4(+) T cells. The antibody response is independent of CD8(+) T cells but requires the presence of CD3(+) cells carrying the NK1.1 markers. PMID- 12573586 TI - Isolation and characterisation of the rabies virus N degrees-P complex produced in insect cells. AB - When the nucleoprotein (N) of nonsegmented negative-strand RNA viruses is expressed in insect cells, it binds to cellular RNA and forms N-RNA complexes just like viral nucleocapsids. However, in virus-infected cells, N is prevented from binding to cellular RNA because a soluble complex is formed between N and the viral phosphoprotein (P), the N degrees -P complex. N is only released from this complex for binding to newly made viral or complementary RNA. We coexpressed rabies virus N and P proteins in insect cells and purified the N degrees -P complex. Characterisation by gel filtration, polyacrylamide gel electrophoresis, analytical ultracentrifugation, native mass spectroscopy, and electron microscopy showed that the complex consists of one N protein plus two P proteins, i.e., an N degrees -P(2) complex. PMID- 12573587 TI - Phenotypic analysis of peripheral blood gammadelta T lymphocytes and their targeting by human immunodeficiency virus type 1 in vivo. AB - There is increasing evidence that a wider range of lymphoid cell types other than CD4(+) T helper lymphocytes are infected with HIV-1 in vivo, including CD8 lymphocytes, natural killer cells, and reticulodendritic cells. Each potentially contributes to the reservoir of infected cells that resist antiviral treatment and to the impairment of immune responses in AIDS. By quantitative PCR for HIV proviral sequences we have now obtained evidence for substantial infection of gammadelta lymphocytes, contributing 3-45% of the proviral load in peripheral blood. A large proportion of gammadelta lymphocytes constitutively expressed the chemokine receptors CCR5 and CXCR4, with evidence for marked up-regulation of CD8 in samples from HIV-infected individuals, corresponding to an activated phenotype. That gammadelta lymphocytes might be susceptible to HIV infection was investigated using in vitro infectivity assays of recombinant HIV-expressing green fluorescent protein, followed by flow cytometry. gammadelta, CD4, and CD8 lymphocytes were each productively infected, with gammadelta lymphocytes showing the greatest susceptibility. For each cell type, blocking assays with an anti-CD4 monoclonal antibody indicated that entry was CD4-dependent. PMID- 12573588 TI - CD4-independent protective cytotoxic T cells induced in early life by a non replicative delivery system based on virus-like particles. AB - The relative immaturity of the neonatal immune system limits CD4(+) Th1 and cytotoxic T lymphocyte (CTL) responses, and represents a significant challenge for the development of vaccines against intracellular pathogens. In this report, we demonstrate the ability of a non-replicative delivery system based on parvovirus-like particles (VLP) to induce CTL responses in the neonatal period. A single immunization of 1-week-old BALB/c mice with recombinant VLP carrying a CD8(+) T cell determinant from lymphocytic choriomeningitis virus (VLP-LCMV) induced antigen-specific CD8(+) cytotoxic T cells that were similar to those elicited by adult immunization, as assessed by cytotoxic activity, interferon (IFN)-gamma secretion, cytotoxic precursor cell frequencies, in vitro avidity for antigen and protective activity against viral challenge. These CTL responses are elicited within 2 weeks of a single immunization, in the absence of adjuvant and independently of the presence and help of CD4(+) T cells, highlighting the potential of VLP as candidate vaccine vectors in early life. PMID- 12573589 TI - Recovery of infectivity from cDNA clones of nodamura virus and identification of small nonstructural proteins. AB - Nodamura virus (NoV) was the first isolated member of the Nodaviridae, and is the type species of the alphanodavirus genus. The alphanodaviruses infect insects; NoV is unique in that it can also lethally infect mammals. Nodaviruses have bipartite positive-sense RNA genomes in which RNA1 encodes the RNA-dependent RNA polymerase and the smaller genome segment, RNA2, encodes the capsid protein precursor. To facilitate the study of NoV, we generated infectious cDNA clones of its two genomic RNAs. Transcription of these NoV1 and NoV2 cDNAs in mammalian cells led to viral RNA replication, protein synthesis, and production of infectious virus. Subgenomic RNA3 was produced during RNA replication and encodes nonstructural proteins B1 and B2 in overlapping ORFs. Site-directed mutagenesis of these ORFs, followed by SDS-PAGE and MALDI-TOF mass spectrometry analyses, showed synthesis of B1 and two forms of B2 (B2-134 and B2-137) during viral replication. We also characterized a point mutation in RNA1 far upstream of the RNA3 region that resulted in decreased RNA3 synthesis and RNA2 replication, and a reduced yield of infectious particles. The ability to reproduce the entire life cycle of this unusual nodavirus from cDNA clones will facilitate further analysis of NoV RNA replication and pathogenesis. PMID- 12573590 TI - Identification of a minimal sequence required for activation of the tomato golden mosaic virus coat protein promoter in protoplasts. AB - Transient expression studies using a Nicotiana benthamiana suspension cell derived protoplast system have identified a minimal sequence that is necessary and sufficient for activation of the tomato golden mosaic virus coat protein (CP) promoter by the viral TrAP protein (also called AL2). The sequence has a bipartite arrangement in which elements located between -125 to -107 and -96 to 60 from the transcription start site are both required for TrAP-mediated activation. One of the sequences (-96 to -60) also appears to interact with a repressor, as its deletion increases basal promoter activity in the absence of TrAP. That competition experiments using the -107 to -60 sequence to titrate the repressor also resulted in increased basal transcription is consistent with this idea. Thus, in a protoplast system which models mesophyll, regulation of the minimal CP promoter involves both activation and derepression by TrAP. PMID- 12573591 TI - Individual expression of sindbis virus glycoproteins. E1 alone promotes cell fusion. AB - The envelope of alphavirus particles contains two major glycoproteins, E1 and E2, that participate in virus entry and assembly of new virus particles. Interactions between these glycoproteins determine their correct functioning. The expression of each glycoprotein in the absence of the other counterpart was achieved by means of electroporation of modified Sindbis virus (SV) genomes. In addition, in trans coexpression of both glycoproteins was also tested in BHK cells. Synthesis of the E1 glycoprotein alone gave rise to cell fusion after incubation in low-pH medium. In addition, expression of E1 in the absence of the E2 precursor, PE2 (E3+E2), induced the formation of cytoplasmic vacuoles in the transfected cells. The normal phenotype was recovered when PE2 was coexpressed in trans with E1. Moreover, this coexpression modified the processing of the PE2 glycoprotein. PE2 synthesized in the absence of E1 gave rise to a product, E2', whose migration was slower in SDS-polyacrylamide gel than that of genuine E2 from SV-infected cells. This alteration was corrected upon in trans coexpression of E1 and PE2. These results suggest that the two glycoproteins, E1 and PE2, interact after their expression from two separate SV genomes. Notably, BHK cells cotransfected with the two modified genomes produced SV particles. Our findings suggest that SV E1 and E2 synthesized in trans can interact with each other and participate together with capsid protein in the assembly of new virus particles. PMID- 12573592 TI - Replication, immunogenicity, and protective properties of live-attenuated simian immunodeficiency viruses expressing interleukin-4 or interferon-gamma. AB - Nef deletion mutants of SIV-expressing interleukin-4 (SIV-IL4) or interferon gamma (SIV-IFN) were constructed to study the effect of interferon-gamma (IFN gamma) and interleukin-4 (IL-4) on viral load, immunogenicity, and protective properties. Four rhesus monkeys were infected with SIV-IL4 and four were infected with SIV-IFN. During the acute phase of infection, the cell-associated viral load, but not the plasma viral RNA load, was approximately 10-fold lower in SIV IFN-infected macaques than in SIV-IL4-infected rhesus monkeys. The viral load declined to hardly detectable levels 4 months postinfection in all animals. SIV antibody titers and the affinity of these antibodies were higher in SIV-IL4 infected macaques than in SIV-IFN-infected animals, consistent with a stimulation of T helper cell type 2 immune responses by IL-4. At peak viremia, there was a trend to higher interleukin-12 and perforin mRNA levels of the lymph nodes in the SIV-IFN-infected macaques than in the SIV-IL4-infected monkeys. Deletion of the viral IFN gene, but not the viral IL-4 gene, after the development of antiviral immune responses suggests a repressive effect of IFN, but not IL-4, on virus spread in vivo. A trend to higher set point viral RNA levels in SIV-IL4-infected monkeys in comparison to monkeys infected with the parental nef deletion mutant and similar viral RNA levels during the acute phase of infection suggest that IL 4 expression leads to a slight reduction in the control of virus replication by host immune responses. However, SIV-IL4 and SIV-IFN induced protection against a homologous challenge virus. Subsequent challenge with an SIV-HIV-1 hybrid virus (SHIV) also revealed protection in the absence of neutralizing antibodies. PMID- 12573593 TI - A transcriptional initiator overlaps with a conserved YY1 binding site in the long control region of human papillomavirus type 16. AB - A single promoter has so far been found in the long control region (LCRs) of human papillomavirus-16 (HPV-16). Multiple promoters exist in the LCRs of several other papillomaviruses, which are spliced to become mRNAs for late and some early genes. Here we have investigated whether such promoters exist in the LCR of HPV 16. In in vitro transcription experiments, we detected a strong transcript starting 280 bp downstream from the 3' end of the L1 gene between a nuclear matrix attachment region and the epithelial-specific enhancer. Promoter activity coincides with a GCCATTTT motif, which binds the transcription factor YY1 (YY1 7436). The A of this motif is the first nucleotide of the transcripts and identifies YY1-7436 as an initiator. Genomic segments with YY1-7436 initiate expression of a luciferase reporter gene in transfection experiments. Mutational analysis of YY1-7436 suggests, however, that promoter function originates from another factor but YY1, which can contact overlapping sequences. Promoter activity of YY1-7436 is modulated by upstream A-T-rich sequences, which bind the basal transcription factor TFIID, and it is stimulated by the viral E2 protein binding to a downstream E2 binding site. In differentiating W12 cells, which contain episomal HPV-16 copies, we detected transcripts including LCR sequences downstream of YY1-7436, which were differentially spliced to early and late genes. However, we could not detect 5' ends mapping to YY1-7436, but we detected two novel HPV-16 promoters within the L1 gene. Conservation of the arrangement of the YY1 and E2 binding sites suggests a role in important biological functions, which, however, is difficult to confirm in every type of cell culture. The study of W12 cells complements the examination of YY1-7436 and points to yet undetected promoters upstream of the LCR. PMID- 12573595 TI - Veterinary pharmacology: where to in the 21st century? PMID- 12573596 TI - Antibiotic treatment of animals--a different approach to rational dosing. PMID- 12573597 TI - The management of animal pain where have we been, where are now, and where are we going? PMID- 12573598 TI - Pharmacokinetics and PK-PD modelling of danofloxacin in camel serum and tissue cage fluids. AB - The pharmacokinetics and pharmacodynamics of danofloxacin were studied in the camel in a two period cross-over study. After intravenous (i.v.) administration at a dose rate of 1.25 mg/kg, the pharmacokinetics of danofloxacin indicated a high volume of distribution (V(d(area))=3.43 L/kg), relatively rapid clearance (0.44 L/kg/h) and half-life of 5.37 h. After intramuscular (i.m.) dosing absorption was complete (F=114.5) and rapid (T((1/2)abs)=0.12 h) and terminal half-life was 5.71 h. Danofloxacin penetrated fairly slowly into both inflamed (exudate) and non-inflamed (transudate) tissue cage fluids and was cleared slowly from these fluids, elimination half-life being at least twice that for serum for both exudate and transudate after both i.v. and i.m. dosing. The antibacterial actions of danofloxacin against the camel pathogen Escherichia coli 0157-H7 were determined by measurement of minimum inhibitory concentration (MIC) in vitro (single measurement) and ex vivo measurements of bacterial count at nine times between one and 48 h after i.m. dosing in each of the fluids, serum, exudate, and transudate. Using in vitro MIC data and in vivo pharmacokinetic parameters, the surrogate markers of antimicrobial activity, C(max)/MIC, AUC/MIC and T>MIC, were determined for all three fluids. The ex vivo serum AUC(24 h)/MIC data were integrated with reduction in bacterial count to provide values producing a bacteriostatic action (no change in bacterial count), inhibition of bacterial count by 50%, reduction in bacterial count by 99.9% (bactericidal action) and elimination of bacteria. Mean AUC(24h)/MIC values were 17.20, 20.07, 21.24, and 68.37 h, respectively. To describe the latter, the introduction of a new term to supplement MIC and minimum bactericidal concentration (MBC) is proposed, namely minimum elimination concentration (MEC). A novel means of designing antimicrobial drug dosage schedules for evaluation in clinical trials is proposed, using ex vivo AUC(24h)/MIC values for bactericidal activity and elimination of bacteria together with MIC(90) data for camel pathogens. PMID- 12573599 TI - Comparison of two sampling techniques for the detection of Malassezia pachydermatis on the skin of dogs with chronic dermatitis. AB - Adhesive tape strip and dry swab sampling techniques were compared for the detection of Malassezia pachydermatis on the skin of dogs with chronic dermatitis. One hundred and four dogs were sampled by each of the techniques. Two methods, a culture method and a stain method, were used to assess the sampling techniques. By the adhesive tape strip sampling technique, M. pachydermatis was detected on 83 (80%) dogs using the culture method and on 45 (43%) dogs using the stain method. By the dry swab sampling technique, M. pachydermatis was detected on 55 (53%) dogs using the culture method and on 33 (32%) dogs using the stain method. The study showed that the adhesive tape strip sampling technique, using the culture method, detected Malassezia on the skin of significantly more dogs (P<0.001) than the same technique using the stain method and also significantly more than the dry swab sampling technique, using either the culture or stain methods. It was also shown that an adhesive tape sample could be used to transfer cells to a slide for staining and microscopy prior to being used for culturing Malassezia. PMID- 12573600 TI - Transmission of bovine viral diarrhoea virus by unhygienic vaccination procedures, ambient air, and from contaminated pens. AB - Knowing how bovine viral diarrhoea virus (BVDV) infection spreads via indirect contacts is required in order to plan large-scale eradication schemes against BVDV. In this study, susceptible calves were exposed to BVDV by an unhygienic vaccination procedure, by ambient air and from contaminated pens. Primary BVDV infection was observed in two calves vaccinated with a vaccine against Trichophyton spp that had been contaminated by smearing nasal secretion from a persistently infected (PI) calf on the rubber membrane and penetrating it twice with a hypodermic needle. Four other calves, housed in pairs in two separate housing units near a PI calf for one week--at distances of 1.5 and 10 m, respectively--became infected without having direct contact with the PI calf. Furthermore, two of the three calves housed in a pen directly after removal of a PI calf, but without the pen being cleaned and disinfected, also contracted primary BVDV infection, whereas two calves that entered such a pen four days after removal of another PI calf, did not. In herds where most animals are seronegative to BVDV, indirect airborne transmission of BVDV or contact with a contaminated housing interior may be an important factor in spreading of the virus, once a PI animal is present. However, the spreading of BVDV within herds can be stopped by identifying and removing PI animals and also by ensuring that susceptible breeding animals do not become infected during this procedure. In contrast, injectables contaminated with BVDV may prove to be a significant vector for spreading the infection, not only within an infected herd but, most importantly, also between herds. In our opinion, it is questionable whether medicine bottles, once opened and used within an infected herd, should be used in other herds. In any case, prior knowledge of a herd's BVDV status will help practising veterinarians and technicians to undertake appropriate hygienic measures. PMID- 12573601 TI - Pain alleviation in animals: attitudes and practices of Finnish veterinarians. AB - A questionnaire was sent to 911 Finnish veterinarians to assess their attitudes and practices to pain relief in animals. Responses to statements about recognition and treatment of pain were either to agree or to disagree. The pain caused by specified surgical and clinical conditions was rated. Inquiries were also posed about the number of analgesics available and their use in specific surgical procedures and clinical situations. The questionnaires were returned by 441 respondents. Women and younger veterinarians generally rated pain higher and treated it more frequently than men and older colleagues. Younger veterinarians and those in larger practices also had more analgesics available than older veterinarians and those in smaller practices. Respondents agreed with the statement that relieving pain is beneficial for animals. However, large differences were present in the frequency of use of pain alleviation between different animal species undergoing similar operations and between clinical conditions scored equally in the numerical rating of pain. The severity and clinical relevance of feline pain is probably often underestimated, as cats were less likely to receive analgesics than dogs after similar operations. PMID- 12573602 TI - Differentiation of Setaria digitata and Setaria labiatopapillosa using molecular markers. AB - 5S rRNA intergenic regions of Setaria digitata and Setaria labiatopapillosa were PCR amplified with primers designed from the 5S rRNA gene of Brugia malayi. The ladder-like banding patterns obtained for the amplifications were distinctly different for the two species. Four amplified products were cloned into the pBS vector and completely sequenced. DNA clones from two individual samples of S. digitata, Sd4 and Sd6, showed 97% sequence homology to each other. All sequenced clones showed the presence of the spliced leader (SL) RNA gene with a 22 nucleotide spliced leader sequence. The phylogenetic tree constructed using these data and the 5S rRNA intergenic regions of several other filarial nematodes showed the Setaria species sharing a branch with Dirofilaria. RAPD-PCR analyses identified 107 bands of which 86 were polymorphic (80%). A dendrogram constructed for S. digitata and S. labiatopapillosa separated the two species into two distinct clusters. The polymorphic loci identified by the RAPD-PCR analyses can be studied further to develop species-specific probes/PCR primers for the identification of each species. PMID- 12573603 TI - Strategic administration of enrofloxacin in poultry to achieve higher maximal serum concentrations. AB - To achieve a higher maximal serum concentration (Cs(max)) of enrofloxacin after oral administration of 10mg/kg/day of three commercial preparations of enrofloxacin to chickens, two concentrations of the drug were tested (0.1 and 0.2%), under controlled laboratory conditions. A single oral bolus dose was delivered directly into the proventriculus of each of 240 chickens, which were equally divided into six groups: three received the customary concentration (0.1%), and three received the higher concentration. A quantitative/qualitative microbiological analytical method to determine serum concentrations of enrofloxacin and a software program to obtain pharmacokinetic variables, revealed that time vs. concentration relationships best fitted double peak shape curves, Cs(max1) and Cs(max2). Statistically significant (P>0.01) increments were obtained when 0.2% enrofloxacin oral solutions from the three different commercial preparations were administered. The increments ranged from 175% to 338% for Cs(max1) and 69% to 342% for Cs(max2). Optimal bactericidal concentrations of enrofloxacin are usually twice the value of their minimal inhibitory concentration. Although clinical trials are now required, it would appear that increments in the serum concentration of enrofloxacin may reduce to the rate at which bacterial resistance occurs and so increase clinical efficacy without affecting the cost per treatment. PMID- 12573604 TI - Flea bite hypersensitivity: new aspects on the involvement of mast cells. AB - A study was performed to test the effect of sensitization to flea antigen, followed by exposure to fleas on mast cells (MCs), their subtypes, and IgE+ cells. Biopsies were taken from flea-sensitized dogs (n=28) and non-sensitized dogs (n=5) that had been exposed to fleas. Control groups consisted of flea sensitized (n=12) and non-sensitized dogs (n=9) that were not exposed to fleas. Biopsies, taken before, 24 and 72 h after local flea exposure, were stained with haematoxylin and eosin (H&E), toluidine blue, a double labelling technique for MC chymase and tryptase and anti-IgE. An intradermal test for flea antigen was performed and serum titres of allergen-specific IgE and IgG were measured. Significantly higher numbers (P<0.001) of double labelled MCs compared to toluidine blue stained MCs were detectable in flea-sensitized dogs independent of flea exposure. In contrast, in non-sensitized dogs, the number of toluidine blue stained MCs and the number of double labelled MCs did not differ. In flea sensitized dogs after flea exposure the percentage of C-MC was significantly increased at day 1 (P<0.001) and day 3 (P<0.001), whereas the percentage of TC MCs decreased significantly at day 1 (P<0.001) and day 3 (P<0.05). The percentage of T-MCs decreased (P<0.05 day 0 versus day 1; P<0.05 day 0 versus day 3). No significant difference was detectable after toluidine blue staining and staining for IgE+ cells between the groups nor between the MC density and the number of IgE+ cells. All flea-sensitized dogs had positive skin tests to flea antigen and high serum titres of flea-specific serum IgE and IgG antibodies. In non sensitized dogs, these results were negative. Our data provide strong evidence for an upregulation of MC proteases during the process of sensitization and a generalized selective release of mast cell tryptase after exposure to the antigen. PMID- 12573605 TI - Identification of microsatellites in expressed muscle genes: assessment of a desmin (CT) dinucleotide repeat as a marker for meat quality. AB - We investigated the presence of dinucleotide repeats (microsatellites) in a porcine skeletal muscle cDNA library. In total, microsatellites were found in nearly 4% of all muscle cDNA clones, of which CT- and TG-repeats were the most prevalent. We found a near full-length desmin cDNA clone with a CT-repeat sited downstream from its stop codon. From 44 commercially reared pigs, six allelic length polymorphisms were identified at this microsatellite locus (128, 131, 134, 135, 136, and 138 bp), whose desmin genotypes were associated with pH change after slaughter (P=0.031) and colour variation of meat (P<0.001 for hue, P<0.005 for lightness, and P=0.014 for b colour). Two of the genotypes (131/131 bp and 131/135 bp) were consistent with paler meat. This finding indicates the usefulness of the cDNA library approach to generate markers for marker-assisted selection and suggests that the desmin microsatellite could be a possible selection marker. PMID- 12573606 TI - Attempts to separate female Ascaris suum antigen and to investigate its partial characterization. AB - The location and separation of Ascaris suum antigen for serological testing was investigated. The antigenic constituent was rich in the ovary of the adult worm and was obtained by dialysis with 50% ammonium sulphate saturated solution. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting analysis demonstrated that the heat labile antigenic preparation showed one major and seven faint bands. The major band seemed also to be a glycoprotein. The sera from pigs with/without hepatic milk spot showed relatively high precipitation titres, while, those from the specific pathogen free pigs manifested low titres. PMID- 12573607 TI - N-ras mutations in canine malignant melanomas. PMID- 12573608 TI - Comparison of antibiotic resistance for Escherichia coli populations isolated from groups of diarrhoeic and control calves. PMID- 12573609 TI - Dermatitis associated with Dirofilaria repens microfilariae in a dog from Rome. PMID- 12573611 TI - Should gene therapy be used for newborns with hemophilia? PMID- 12573612 TI - RNA interference: applications in vertebrates. PMID- 12573613 TI - Global non-viral gene transfer to the primate brain following intravenous administration. AB - Expression plasmids encoding either luciferase or beta-galactosidase were encapsulated in the interior of an "artificial virus" comprised of an 85 nm pegylated immunoliposome, which was targeted to the rhesus monkey brain in vivo with a monoclonal antibody (MAb) to the human insulin receptor (HIR). The HIRMAb enables the liposome carrying the exogenous gene to undergo transcytosis across the blood-brain barrier and endocytosis across the neuronal plasma membrane following intravenous injection. The level of luciferase gene expression in the brain was 50-fold higher in the rhesus monkey as compared to the rat. Widespread neuronal expression of the beta-galactosidase gene in primate brain was demonstrated by both histochemistry and confocal microscopy. This approach makes feasible reversible adult transgenics in 24 hours. PMID- 12573614 TI - Enhancement of adenovirus-mediated gene transfer to the airways by DEAE dextran and sodium caprate in vivo. AB - Gene transfer to the trachea and airways by adenoviral vectors is limited by the basolateral localization of viral receptors, resulting in relatively low levels of transduction. Modification of paracellular permeability by sodium caprate, which opens tight junctions, enhances gene transfer from the apical side of cultured human airway epithelial cells. Based on this observation we investigated whether Na-caprate could also increase gene transfer when applied to the luminal surface of the airway epithelia in vivo and compared these results with EGTA, which has previously been shown to enhance adenovirus transduction. Transgene expression in the trachea and upper airways was increased 25-fold by a 10-min pretreatment with 50 mM Na-caprate, corresponding to a 3-fold improvement over EGTA. In the more peripheral airways EGTA had no effect, whereas expression of beta-gal was increased 3-fold by Na-caprate. When the adenovirus was complexed with DEAE dextran, transduction of the airway epithelia after Na-caprate pretreatment was increased 45-fold over virus alone. In conclusion, Na-caprate facilitates gene transfer to airway epithelia, particularly when adenovirus is complexed with DEAE dextran, and may in future be used in a clinical setting to enhance the efficiency of vectors for gene therapy of cystic fibrosis via airway delivery. PMID- 12573615 TI - In vivo transduction of photoreceptors or ciliary body by intravitreal injection of pseudotyped adenoviral vectors. AB - Strategies for retargeting adenoviral (Ad) vectors have been developed, but their in vivo efficacy remains to be demonstrated. Gene delivery to specific ocular cell types represents an approach to treating many diseases that cause irreversible blindness. One of these cell types, the photoreceptor (PR), is not infected by standard Ad5-based vectors. We evaluated gene delivery after intraocular injection of Ads pseudotyped with three different fiber proteins and found three distinct patterns of infection. An intravitreally injected Ad5 vector readily infected the iris, corneal endothelium, and ciliary body, while few cells in the retina expressed transgene product. In contrast, an Ad3-pseudotyped virus selectively transduced ciliary body, of interest for treating diseases such as glaucoma. A vector pseudotyped with the fiber protein of Ad37 transduced PRs as well as ciliary body. This finding has potential application to the treatment of retinal degenerative or neovascular diseases. These studies demonstrate cell type selective gene delivery in vivo with retargeted Ads, provide information about the cellular tropisms of several Ad serotypes, and should lead to improved strategies for preserving vision. PMID- 12573616 TI - Selective depletion or blockade of Kupffer cells leads to enhanced and prolonged hepatic transgene expression using high-capacity adenoviral vectors. AB - Tissue macrophages, in particular hepatic Kupffer cells (KCs), contribute to early inflammatory responses following adenoviral vector administration. This study evaluates the effect of selective and transient (3 days) depletion of KCs by a single injection of clodronate liposomes on the in vivo performance of high capacity adenoviral (HC-Ad) vectors. In KC-depleted C57BL/6 and C3H mice increased and stabilized hAAT levels were observed following intravenous injection of HC-Ad vectors expressing human alpha-1 anti-trypsin (hAAT) either from the hAAT promoter or from the human cytomegalovirus promoter. Comparable increases in hAAT levels were obtained in mice preinjected with a transcriptionally silent HC-Ad vector. Interestingly, in the majority of animals of both strains depletion of KCs was sufficient to prevent the generation of anti hAAT antibodies, resulting in prolonged transgene expression. Thus, short-term and selective depletion of hepatic macrophages at the same time significantly increased hepatic transgene expression and reduced the humoral immune response to the transgenic protein. PMID- 12573617 TI - Adeno-associated viral vector-mediated gene transfer of VEGF normalizes skeletal muscle oxygen tension and induces arteriogenesis in ischemic rat hindlimb. AB - Critical limb ischemia is an important clinical problem that often leads to disability and limb loss. Vascular endothelial growth factor (VEGF), delivered either as recombinant protein or as gene therapy, has been shown to promote both collateral artery formation (arteriogenesis) and capillary angiogenesis in animal models of hindlimb ischemia. However, none of the previous studies has demonstrated an improvement in tissue hypoxia, the condition that drives the molecular response to ischemia. Furthermore, the optimal vector and route of gene delivery have not been determined. Recently, adeno-associated viral (AAV) vectors, which efficiently transduce skeletal muscle and produce sustained transgene expression, have been used as gene therapy vectors. We asked whether an intra-arterial injection of AAV-VEGF(165) normalizes muscle oxygen tension by increasing skeletal muscle oxygen tension, and promotes arteriogenesis and angiogenesis in a rat model of severe hindlimb ischemia. We found that AAV-VEGF treatment normalized muscle oxygen tension in the ischemic limb. In contrast, vehicle and AAV-lacZ-treated limbs remained ischemic. Collateral arteries were more numerous in AAV-VEGF-treated rats, but, surprisingly, capillaries were not. We conclude that intra-arterial AAV-mediated gene transfer of AAV-VEGF(165) normalizes muscle oxygen tension and leads to arteriogenesis in rats with severe hindlimb ischemia. PMID- 12573618 TI - Biodistribution and efficacy of donor T lymphocytes in a murine model of lysosomal storage disease. AB - Lymphocyte-directed gene transfer has been proposed as potential therapy to treat certain congenital immunological deficiencies as well as other genetic diseases such as lysosomal storage diseases (LSDs). To understand better the extent to which adoptively transferred peripheral T lymphocytes (PTLs) are able to ameliorate LSDs we utilized the beta-glucuronidase-deficient mouse as a model system. PTLs (1 x 10(7)) isolated from the spleen of syngeneic mice overexpressing ( approximately 8-fold) human beta-glucuronidase (GUSB) were injected intravenously into young adult beta-glucuronidase-deficient mice without myeloablative conditioning. Using biochemical and histochemical assays, we were able to track the donor lymphocytes in vivo. Donor lymphocytes were detected in relatively high numbers in liver, spleen, small intestine, mesenteric lymph node, and thymus for at least 5 months, the last time point of analysis. Although liver and spleen had the highest total GUSB activity, histopathologic analysis demonstrated minimal to no correction of lysosomal distention at all time points studied. By contrast, we have shown in earlier studies that administration of similar numbers of macrophages reduced lysosomal storage in several organs, including liver and spleen. To understand this difference in efficacy, we compared the relative level of GUSB released into the medium by nonactivated and activated PTLs as well as by macrophages. Macrophages released >50-fold excess enzyme compared to either activated or nonactivated PTLs. These data suggest that a LSD can be more effectively treated by directing a gene therapy approach to a hematopoietic lineage other than T lymphocytes. PMID- 12573619 TI - An oncolytic measles virus engineered to enter cells through the CD20 antigen. AB - We have earlier shown that attenuated measles virus (MV) has therapeutic potential as a replicating oncolytic virus in models of non-Hodgkin's lymphoma (NHL). In the current study, we investigated whether we could obtain replicating MVs capable of entering CD20(+) target cells through an interaction between a single-chain (scFv) anti-CD20 antibody and the CD20 antigen, a target of considerable clinical relevance in NHL. We replaced the H envelope glycoprotein of MV by an H-scFv anti-CD20 fusion protein with and without a protease-cleavable linker. Biochemical analysis of purified virions confirmed that the modified H proteins were incorporated into the viral particles with efficiency similar to unmodified H. Experiments employing CHO cells and CHO cells expressing human CD20 indicated that the MVH alpha CD20 viruses were able to replicate well in CHOCD20 but not CHO cells. MVH alpha CD20 or a nonmodified control MV were administered systemically to immunodeficient mice bearing bilateral human tumor xenografts, one side with and the other side without CD20 expression. Growth of CD20(+) tumors was retarded by MVH alpha CD20 as compared with the control virus. The viruses had equivalent effects on the CD20(-) tumors. Thus we have demonstrated that the entry of a replicating oncolytic virus can be mediated through an interaction between a highly clinically relevant single-chain antibody and its target antigen, and we have shown that this interaction enhances in vivo oncolytic activity. PMID- 12573620 TI - Lactosylated poly-L-lysine targets a potential lactose receptor in cystic fibrosis and non-cystic fibrosis airway epithelial cells. AB - Poly-L-lysine with 40% of the epsilon -amino groups substituted with lactosyl residues facilitated the internalization of lactosylated poly-L-lysine/cDNA complexes into cystic fibrosis (CF) and non-CF airway epithelial cells. It was previously shown that lactosylated poly-L-lysine enhanced the transfer of cDNA into the cell nucleus, resulting in transfection. The cell entry of lactosylated poly-L-lysine/cDNA complexes, however, has not been elucidated and we hypothesized that entry of the complex was by receptor-mediated endocytosis. It is shown here that binding of the vector/cDNA complexes to the cell membrane was inhibited by lactose but not N-acetyl glucosamine. Examination by electron microscopy revealed the complexes in clathrin-coated pits. Furthermore, the complexes colocalized with transferrin during cell entry and were shown in early endosomes. These results demonstrated that lactosylated poly-L-lysine/cDNA complexes enter airway epithelial cells via receptor-mediated endocytosis utilizing lactose-binding receptors, which employ the clathrin-coated pit for internalization. Taken together with the fact that nuclear translocation also is enhanced by lactose, these results demonstrate why lactosylated poly-L-lysine is an excellent vector for transfection of airway epithelial cells. Moreover, other carbohydrates covalently linked to poly-L-lysine for targeting other specific cell types, combined with lactosyl residues, can be designed for the development of other molecular conjugates for gene transfer. PMID- 12573621 TI - Free cationic liposomes inhibit the inflammatory response to cationic lipid-DNA complex injected intravenously and enhance its transfection efficiency. AB - In this report, we show that intravenous (i.v.) injection into mice of a complex made of the cationic lipid diC14-amidine and the luciferase reporter plasmid (pCMV-luc) results in efficient gene expression in several organs but elicits an inflammatory response characterized by a release of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) into the serum of treated animals. A single preinjection of free diC14-amidine liposomes improves the i.v. transfection efficiency of the diC14-amidine/protamine/pCMV-luc complex as much as 40 times. This improvement is correlated with the ability of free liposomes to inhibit TNF-alpha but not IFN-gamma production resulting from complex injection. TNF-alpha-rich serum obtained from mice injected with diC14 amidine/protamine/pCMV-luc complex inhibits luciferase expression in transfected mouse lung endothelial (MLE) cells cultured in vitro, whereas IFN-gamma has no effect. This inhibitory effect can be partly abolished by treating the mouse serum with a specific anti-TNF-alpha antibody. These data point out that cationic lipids are potent inhibitors of the inflammatory response to the CpG motifs in plasmid DNA. This property is shown to enhance the in vivo transfection efficiency. PMID- 12573622 TI - Cationic lipid and polymer-based gene delivery to human pancreatic islets. AB - Transplantation of pancreatic islets has great potential for treating Type I diabetes. Ex vivo gene therapy may promote re-vascularization or inhibit apoptosis of the islets and promote graft. In this study, we investigated the feasibility of non-viral gene delivery using Enhanced Green Fluorescent Protein (EGFP) and human Vascular Endothelial Growth Factor (hVEGF(165)) expression plasmids as model reporter and therapeutic genes. LipofectAMINE/pDNA and Superfect/pDNA complexes showed high transfection efficiency in rapidly dividing Jurkat cells, but low transfection in non-dividing human islets. LipofectAMINE/pCAGGS-hVEGF transfected islets showed relatively higher levels of hVEGF than in those transfected with LipofectAMINE/pCMS-EGFP complexes or 5% glucose. To exclude endogenously secreted hVEGF, real time RT-PCR experiment was repeated using pCAGGS vector-specific forward primer and hVEGF gene-specific reverse primer. In this case, both non-transfected islets and the islets transfected with LipofectAMINE/pCMS-EGFP complexes showed negligible amplification of hVEGF. On glucose challenge, insulin release from LipofectAMINE/pCAGGS-hVEGF transfected human islets increased from 10.78 +/- 4.56 to 65 +/- 5 ng/ml, suggesting little adverse effect on islet beta cell response to glucose challenge. The low transfection efficiency is due to the islets being a cluster of approximately 1000 non-dividing cells. This underscores the importance of experimentation with the actual human islets. PMID- 12573623 TI - Helper-independent and AAV-ITR-independent chromosomal integration of double stranded linear DNA vectors in mice. AB - Nonviral plasmid DNA is a promising vector for achieving ex vivo and in vivo gene transfer. However, transgene expression is usually transient, especially in dividing target cells due to loss of vector genomes. Here we describe the use of naked double-stranded (ds) linear DNA as a way to insert exogenous DNA sequences into chromosomes of mouse hepatocytes in vivo, without helper components such as integrase or transposase. We constructed ds linear DNA vectors with or without adeno-associated virus inverted terminal repeats (AAV-ITRs), introduced them into mouse hepatocytes in vivo using a hydrodynamics-based transfection technique, and analyzed for vector genome integration in various ways. Surprisingly, these linear DNA molecules integrated in mouse hepatocytes in vivo at a level of 0.3 0.5 vector genome, or more, per diploid genomic equivalent irrespective of the AAV-ITR sequences. Our results establish a novel and simple way to engineer chromosomes in vivo and provide further insights into the mechanisms of recombinant AAV vector integration in vivo. In addition, they may provide a clue for developing new nonviral integrating gene delivery vector systems. PMID- 12573624 TI - Free DNA ends are essential for concatemerization of synthetic double-stranded adeno-associated virus vector genomes transfected into mouse hepatocytes in vivo. AB - Recombinant adeno-associated virus (rAAV) vectors stably transduce hepatocytes in vivo. In hepatocyte nuclei, the incoming single-stranded (ss) vector genomes are converted into various forms of double-stranded (ds) genomes including extrachromosomal linear and circular monomers and concatemers, and a small portion of the vector genomes integrate into chromosomes. The mechanism of genome conversion is not well understood. In the present study, we analyzed the role of inverted terminal repeat (ITR) sequences of ds circular or linear rAAV vector intermediates in concatemerization. We synthesized supercoiled ds circular monomers with a double-D ITR (DDITR) (C+), and ds linear monomers with an ITR at each end (L+), and their control molecules, C- and L-, which lack the ITR-derived sequences, and transfected mouse hepatocytes with these molecules in vivo to assess their capacity for concatemerization. The transfected L+ or L-, but not C+ or C- molecules, concatemerized in vivo irrespective of the presence or absence of the ITRs. In addition, our results suggested that transfected C+ or C- species were not efficient substrates for integration. Based on these observations, we propose a model whereby ds linear molecules with free DNA ends, but not circular molecules, play an important role in rAAV vector genome concatemerization. PMID- 12573625 TI - Quantification of adeno-associated virus particles and empty capsids by optical density measurement. AB - We show here that UV absorbance of denatured adeno-associated virus (AAV) vector provides a simple, rapid, and direct method for quantifying vector genomes and capsid proteins in solution. We determined the molar extinction coefficients of capsid protein to be 3.72 x 10(6) M(-1) cm(-1) at 260 nm and 6.61 x 10(6) M(-1) cm(-1) at 280 nm. For recombinant AAV vectors, extinction coefficients can be calculated by including the predicted absorbance of the vector DNA. Since the amount of empty capsids in purified vector preparations lowers the A(260)/A(280) ratio in a predictable manner, the vector genome (vg) and capsid particle (cp) titers in purified AAV vector preparations can be calculated from the absorbance at 260 nm and the A(260)/A(280) ratio. To validate this method, the vg and cp titers calculated by UV absorbance were compared with titers determined by quantitative (Q)-PCR and capsid ELISA. The vg titers determined by absorbance agreed well with titers determined by Q-PCR. The cp/vg ratio determined by the A(260)/A(280) method also correlated well with those determined by AAV capsid ELISA in conjunction with Q-PCR. This new method provides a simple and rapid means to determine AAV vg titers and the ratio of empty to full particles in purified virus preparations. PMID- 12573626 TI - Rapid identification of efficient target cleavage sites using a hammerhead ribozyme library in an iterative manner. AB - A major limitation to the effectiveness of ribozymes is definition of accessible sites in targeted RNAs. Although library selection procedures have been developed, they are generally difficult to perform and have not been widely employed. Here we describe a selection technology that utilizes a randomized, active hammerhead ribozyme (Rz) library in an iterative manner. After two rounds of binding under inactive conditions, the selected, active Rz library is incubated with target RNA, and the sites of cleavage are identified on sequencing gels. We performed this library-selection protocol using human papillomavirus type 16 E6/E7 mRNA as target and constructed Rz targeted to the identified sites. Rz targeted to sites identified with this procedure were generally highly active in vitro and, more importantly, they were highly active in cell culture, whereas their catalytically inactive counterparts were not. This protocol can be used to identify a set of potential target sites within a relatively short time. PMID- 12573627 TI - Surface chemical studies on sphalerite and galena using extracellular polysaccharides isolated from Bacillus polymyxa. AB - Adsorption, electrokinetic, microflotation, and flocculation studies have been carried out on sphalerite and galena minerals using extracellular polysaccharides (ECP) isolated from Bacillus polymyxa. The adsorption density of ECP onto galena is found to be higher than that onto sphalerite. The adsorption of ECP onto sphalerite is found to increase from pH 3 to about pH 7, where a maximum is attained, and thereafter continuously decreases. With respect to galena, the adsorption density of ECP steadily increases with increased pH. The addition of ECP correspondingly reduces the negative electrophoretic mobilities of sphalerite and galena in absolute magnitude without shifting their isoelectric points. However, the magnitude of the reduction in the electrophoretic mobility values is found to be greater for galena compared to that for sphalerite. Microflotation tests show that galena is depressed while sphalerite is floated using ECP in the entire pH range investigated. Selective flotation tests on a synthetic mixture of galena and sphalerite corroborate that sphalerite could be floated from galena at pH 9-9.5 using ECP as a depressant for galena. Flocculation tests reveal that in the pH range 9-11, sphalerite is dispersed and galena is flocculated in the presence of ECP. Dissolution tests indicate release of the lattice metal ions from galena and sphalerite, while co-precipitation tests confirm chemical interaction between lead or zinc ions and ECP. Fourier transform infrared spectroscopic studies provide evidence in support of hydrogen bonding and chemical interaction for the adsorption of ECP onto galena/sphalerite surfaces. PMID- 12573633 TI - Surfactant effects of chlorpromazine and imipramine on lipid bilayers containing sphingomyelin and cholesterol. AB - The surface-active drugs chlorpromazine (CPZ) and imipramine (IP) have been tested on large unilamellar vesicles composed of phosphatidylcholine (PC), sphingomyelin (SM), and cholesterol (Ch) in different proportions. The well characterized nonionic detergent Triton X-100 (TX) has also been used in parallel experiments. Leakage of vesicular aqueous contents and bilayer solubilization have been measured for each surfactant molecule and vesicle composition. All three surface-active molecules behave in a qualitatively similar way, irrespective of bilayer composition: they induce leakage at concentrations well below their critical micellar concentrations (cmc) and solubilization near the cmc. In these events, the potency of the three surfactants under study increases with decreasing cmc, in the order IP100%) dose to the spinal cord than planned, even though the planned target coverage may not change substantially. With daily CT guidance using the SBF, we showed that the maximal spinal cord dose is ensured to be within 10-15% of the planned value. PMID- 12573746 TI - Meta-analysis of dose-fractionation radiotherapy trials for the palliation of painful bone metastases. AB - PURPOSE: To compare pain relief among various dose-fractionation schedules of localized radiotherapy (RT) in the treatment of painful bone metastases. METHODS AND MATERIALS: A systematic search for randomized trials of localized RT on bone metastases using different dose fractionations was performed using Medline (1966 to February 2001) and other sources. The primary outcomes of interest were complete and overall pain relief. The studies were divided into three groups: comparisons of doses given as a single fraction, single vs. multiple fractions, and comparisons of doses given as multiple fractions. The complete and overall pain responses for studies comparing single vs. multiple fractions were pooled. Exploratory analyses of the dose-response relationship, using the biologic effective dose (alpha/beta = 10), were performed using results from all three groups of trials. RESULTS: Two trials comparing single vs. single, eight trials comparing single vs. multiple, and six trials comparing multiple vs. multiple fractions were included. The complete and overall response rates from studies comparing single-fraction RT (median 8 Gy, range 8-10 Gy) against multifraction RT (median 20 Gy in 5 fractions, range 20 Gy in 5 fractions to 30 Gy in 10 fractions) were homogeneous and allowed pooling of data. Of 3260 randomized patients in seven studies, 539 (33.4%) of 1613 and 523 (32.3%) of 1618 patients achieved a complete response after single and multifraction RT, respectively, giving a risk ratio of 1.03 (95% confidence interval 0.94-1.14; p = 0.5). The overall response rate was in favor of single-fraction RT (1011 [62.1%] of 1629) compared with multifraction (958 [58.7%] of 1631; risk ratio 1.05, 95% confidence interval 1.00-1.11, p = 0.04), reaching statistical significance. However, when the analysis was restricted to evaluated patients alone, the overall response rates were similar for single fraction and multifraction RT, at 1011 (72.7%) of 1391 and 958 (72.5%) of 1321, respectively (risk ratio 1.00; p = 0.9). Exploratory analyses by biologic effective dose did not reveal any dose-response relationship among the fractionation schedules used (single 8 Gy to 40 Gy in 15 fractions). Of the other results and observations reported in the trials, only the re-irradiation rates were consistently different between the treatment arms (more frequent re-irradiation in lower dose arms among trials reporting re irradiation rates). CONCLUSION: Meta-analysis of reported randomized trials shows no significant difference in complete and overall pain relief between single and multifraction palliative RT for bone metastases. No dose-response relationship could be detected by including data from the multifraction vs. multifraction trials. Additional data are needed to evaluate the role of re-irradiation and the impact of RT on other treatment end points such as quality of life. PMID- 12573747 TI - Intensity-modulated radiotherapy following extrapleural pneumonectomy for the treatment of malignant mesothelioma: clinical implementation. AB - PURPOSE: New insight into the extent of the target volume for the postoperative irradiation of malignant pleural mesothelioma as determined during surgery has indicated that standard conformal radiotherapy (IMRT) is not sufficient for curative treatment. We describe a novel technique for implementing intensity modulated radiotherapy (IMRT) to deliver higher doses to treat the full extent of these complex target volumes. METHODS AND MATERIALS: After extrapleural pneumonectomy, 7 patients underwent simulation, treatment planning, and treatment with IMRT to the involved hemithorax and adjacent abdomen. The target volumes encompassed the entire operative bed, including the ipsilateral mediastinum, anterior pleural reflection, and ipsilateral pericardium and the insertion of the diaphragm and crura. These were extensively marked during surgery with radiopaque markers to facilitate target delineation. RESULTS: Setup uncertainty and respiratory-dependent motion were found to be small. Coverage of the planning target volume was very good, with the crus of the diaphragm the most difficult volume to irradiate. The radiation doses to normal structures were acceptable. CONCLUSION: IMRT for treatment of malignant mesothelioma after extrapleural pneumonectomy results in more potentially curative doses to large, complex target volumes with acceptable doses to normal tissues. PMID- 12573748 TI - Secretory activity and cell cycle alteration of alveolar type II cells in the early and late phase after irradiation. AB - PURPOSE: Type II cells and the surfactant system have been proposed to play a central role in pathogenesis of radiation pneumonitis. We analyzed the secretory function and proliferation parameters of alveolar type II cells in the early (until 24 h) and late phase (1-5 weeks) after irradiation (RT) in vitro and in vivo. METHODS AND MATERIALS: Type II cells were isolated from rats according to the method of Dobbs. Stimulation of secretion was induced with terbutaline, adenosine triphosphate (ATP), and 12-O-tetradecanoylphorbol-13-acetate (TPA) for a 2-h period. Determination of secretion was performed using (3)H-labeled phosphatidylcholine. For the early-phase analysis, freshly isolated and adherent type II cells were irradiated in vitro with 9-21 Gy (stepwise increase of 3 Gy). Secretion stimulation was initiated 1, 6, 24, and 48 h after RT. For late-phase analysis, type II cells were isolated 1-5 weeks after 18 Gy whole lung or sham RT. Each experiment was repeated at least fivefold. Flow cytometry was used to determine cell cycle distribution and proliferating cell nuclear antigen index. RESULTS: During the early-phase (in vitro) analysis, we found a normal stimulation of surfactant secretion in irradiated, as well as unirradiated, cells. No change in basal secretion and no dose effect were seen. During the late phase, 1-5 weeks after whole lung RT, we observed enhanced secretory activity for all secretagogues and a small increase in basal secretion in Weeks 3 and 4 (pneumonitis phase) compared with controls. The total number of isolated type II cells, as well as the rate of viable cells, decreased after the second post-RT week. Cell cycle alterations suggesting an irreversible G(2)/M block occurred in the second post-RT week and did not resolve during the observation period. The proliferating cell nuclear antigen index of type II cells from irradiated rats did not differ from that of controls. CONCLUSION: In contrast to literature data, we observed no direct effect of radiation on secretory activity in the early phase after RT. In our study of isolated type II cells, as well as in intact animals, RT did not result in an impaired surfactant secretion up to 5 weeks after RT. Our in vivo experiments even showed an increased response of phosphatidylcholine secretion to all known secretagogues at Weeks 3 and 4 after whole lung RT, possibly due to inflammatory cytokines. Cell cycle alterations with G(2)/M block and cell loss in the late post-RT period may contribute more to the manifestation of radiation-induced lung damage than functional impairment in type II cells. PMID- 12573749 TI - Process of care and preliminary outcome in limited-stage small-cell lung cancer: results of the 1995-1997 patterns of care study in Japan. AB - PURPOSE: To evaluate the practice process using the national average (NA); to compare differences in the process of care by age group; and to provide a preliminary outcome data for limited-stage small-cell lung cancer in Japan. METHODS AND MATERIALS: The Patterns of Care Study conducted a nationwide survey of the care process for Stage I-III small-cell lung cancer in Japan. Patients were divided into three age groups: <65 years (younger group, n = 73); between 65 and 74 years (intermediate group, n = 81); and >or=75 years (elderly group, n = 20). RESULTS: The NA for the total dose was 49.0 Gy, and for use of photon energy >or=6 MV, chemotherapy, and prophylactic cranial irradiation was 77.3%, 93.2%, and 1.69%, respectively. Age stratification had no impact on the variables of radiotherapy (RT) such as total dose and field size. Only 37% of patients received chemotherapy and thoracic RT concurrently. The proportion of patients who received chemotherapy and RT concurrently was 44%, 27%, and 25% of the younger, intermediate, and elderly groups, respectively (p = 0.029). Etoposide and cisplatin were less frequently used in the elderly group (>or=75 years old). Overall survival at 3 years for the entire group was 26%. The 3-year survival rate was 30% in the younger group, 28% in the intermediate group, and 9% in the elderly group. Variables found to have a significant impact on survival by multivariate analysis were the use of chemotherapy (p = 0.030), age (p = 0.032), and T stage (p = 0.042). CONCLUSION: Calculated NAs showed that the results of clinical study had favorably penetrated into the practice process in Japan. The results demonstrated that patient age significantly influenced the process of chemotherapy such as the use of etoposide and cisplatin for limited-stage small cell lung cancer in Japan. More concurrent chemotherapy and thoracic RT and the application of prophylactic cranial irradiation for complete responders need to be investigated in the future. PMID- 12573750 TI - Breast radiotherapy with inclusion of internal mammary nodes: a comparison of techniques with three-dimensional planning. AB - PURPOSE: To compare the partially wide tangent (PWT) technique of breast and internal mammary chain irradiation with photon/electron (P/E) and standard tangent (ST) techniques in terms of dose homogeneity within breast and the dose to critical structures such as the heart and lung. METHODS AND MATERIALS: Sixteen left breast cancer patients underwent CT simulation. The breasts, lungs, heart, and internal mammary chain were contoured and treatment plans generated on a three-dimensional planning system (Helax-TMS). RESULTS: The mean dose to the left breast volume with the ST, P/E, and PWT techniques was 94.7%, 98.4%, and 96.5%, respectively (p = 0.029). The left lung received the lowest mean dose with the ST technique (13.9%) compared with PWT (22.8%) and P/E (24.3%). The internal mammary chain volume was most consistently treated with the PWT (mean dose 99%) vs. P/E (86%) and ST (38.4%) techniques. The heart received the least dose with ST (mean dose 6.7%) vs. PWT (10.3%) and P/E (19%). The PWT treated the greatest amount of contralateral breast (mean dose 5.8%) vs. ST (3.2%) vs. P/E (2.8%). CONCLUSION: The PWT technique treats the internal mammary chain with acceptable toxicity to major organs, especially the heart, and with reasonable dose homogeneity in patients with mastectomy or intact breasts. PMID- 12573751 TI - Evidence-based medicine: its effect on treatment recommendations as illustrated by the changing role of postmastectomy irradiation to treat breast cancer. AB - PURPOSE: To illustrate the effect that the quality of evidence has on clinical practice, we examined how the role of radiotherapy in treating breast cancer has changed over the years as the quality of evidence evolved from anecdotal evidence based on expert opinion to randomized clinical trials and meta-analyses. METHODS: We searched the medical literature for key randomized studies and meta-analyses that have influenced the clinical use of postmastectomy irradiation since the first randomized trials in breast cancer in the 1950s. We discuss how clinical practice changed based on the outcomes of these trials, and then discuss the quality of those trials based on the criteria currently used to assess evidence from randomized trials (CONSORT) and meta-analysis (QUORUM). RESULTS: Evidence published from the early trials and meta-analyses on the role of postmastectomy irradiation had a strong effect on clinical practice. Examination of these studies, however, continues to show significant flaws in trial design that, by today's evidence-based standards, would not meet standards of quality. CONCLUSION: The quality of evidence has a strong effect on shaping clinical practice and needs to be continually assessed. Current guidelines developed to critique both individual randomized trials and meta-analyses are helping to establish high standards for trial design and interpretation. Evidence from older trials that were not guided by well-developed guidelines need to be reviewed, particularly when results from those trials are continually updated and used to generate evidence on which to base current clinical practice. PMID- 12573752 TI - Long-term radiation sequelae after breast-conserving therapy in women with early stage breast cancer: an observational study using the LENT-SOMA scoring system. AB - PURPOSE: To evaluate the long-term toxicity after breast-conserving therapy in women with early-stage breast cancer. METHODS AND MATERIALS: Late toxicity according to the late effects of normal tissue-subjective, objective, management, and analytic (LENT-SOMA) criteria and cosmetic outcome (graded by physicians) were evaluated in 590 of 2943 women with early-stage breast cancer who were irradiated between 1983 and 1995 using the following fractionation schedules: group A, 1983-1987, 2.5 Gy 4x/wk to 60 Gy; group B, 1988-1993, 2.5 Gy 4x/wk to 55 Gy, group C, 1994-1995, 2.0 Gy 5x/wk to 55 Gy. RESULTS: LENT-SOMA Grade 3-4 toxicity was observed as follows: group A (median follow-up 171 months; range 154 222 months), fibrosis 16% (7 of 45), telangiectasia 18% (8 of 45), and atrophy 4% (2 of 45); group B (median follow-up 113 months; range 78-164 months), pain 2% (8 of 345), fibrosis 10% (34 of 345), telangiectasia 10% (33 of 345), arm edema 1% (2 of 345), and atrophy 8% (27 of 345); and group C (median follow-up 75 months, range 51-96 months, n = 200), occurrence of Grade 3-4 late morbidity 100 cc were excluded from the analysis. All patients considered for the analysis (n = 128) were treated at a nominal dose equal to 66.6-70.2 Gy (ICRU dose 68-72.5 Gy; median 70 Gy) with conformal (n = 76) or conventional (n = 52) four-field technique (1.8 Gy/fr). Clinical parameters such as diabetes mellitus, acute rectal bleeding, hypertension, age, and hormonal therapy were considered. Late rectal bleeding was scored using a modified Radiation Therapy Oncology Group scale, and patients experiencing >or=Grade 2 were considered bleeders. Median follow-up was 36 months (range 12-72). Mean and median rectal dose were considered, together with rectal volume and the % fraction of rectum receiving more than 50, 55, 60, and 65 Gy (V50, V55, V60, V65, respectively). Median and quartile values of all parameters were taken as cutoff for statistical analysis. Univariate (log-rank) and multivariate (Cox hazard model) analyses were performed. RESULTS: Fourteen of 128 patients experienced >or=Grade 2 late bleeding (3-year actuarial incidence 10.5%). A significant correlation between a number of cutoff values and late rectal bleeding was found. In particular, a mean dose >or=54 Gy, V50 >or=63%, V55 >or=57%, and V60 >or=50% was highly predictive of late bleeding (p or=63% and those with V50 <63% (DVH grouping), data were fitted with a Cox regression hazard model using DVH grouping, rectal volume, and the main clinical parameters as independent variables. Results of the analysis showed that DVH grouping (relative risk 3.3; p = 0.04) and acute bleeding (relative risk 7.1; p = 0.001) are independently predictive of late bleeding. CONCLUSIONS: DVHs of the rectum are significantly correlated with late bleeding for patients irradiated at 66.6-70.2 Gy after radical prostatectomy. PMID- 12573757 TI - Radiotherapy alone in the treatment of uterine cervix cancer with telecobalt and low-dose-rate brachytherapy: retrospective analysis of results and variables. AB - PURPOSE: This retrospective analysis aims to report results and variables from patients with cervix cancer treated by radiation therapy alone with telecobalt and low-dose-rate brachytherapy (LDRB). METHODS AND MATERIALS: Between September 1989 and September 1995, 190 patients with histologic diagnosis of cervix carcinoma were treated with telecobalt for external beam radiotherapy (EBR), followed by one or two insertions of LDRB. Stage distribution according to patients was the following: IB, 12; IIA, 4; IIB, 105; and IIIB, 69. Median dose of EBR at whole pelvis was 40 Gy, and median parametrial doses for Stages II and III patients were 50 Gy and 60 Gy, respectively. Median doses of LDRB at point A for patients treated with one and two insertions were 38 Gy and 50 Gy, respectively. RESULTS: Median follow-up time was 70 months (range: 8-127 months). Overall survival, disease-free survival, and 5-year local control of patients at Stages I, II, and III were 83%, 78%, and 46%; 83%, 82%, and 49%; and 92%, 87%, and 58%, respectively. Overall incidence of late complications in the rectum, small bowel, and urinary tract was 15.3% (19/190), 4.2% (8/190), and 6.8% (13/190), respectively. The actuarial 5-year rectal, small bowel, and urinary incidence of late complications was 16.1%, 4.6%, and 7.6%, respectively. Clinical stage was the only significant variable for overall 5-year survival (p = 0.001), for disease-free survival (p = 0.001), and for local control (p = 0.001). Stage II patients more than 50 years old had better disease-free survival and local control at 5 years (p = 0.004). None of the analyzed variables influenced the actuarial 5-year incidence of late complications. CONCLUSIONS: Results of this series suggest that the use of telecobalt equipment for EBR with doses up to 50 Gy at whole pelvis, prior to brachytherapy, is an acceptable technique for radiation therapy alone in the treatment of cervix cancer, especially in developing countries, including Brazil, where telecobalt machines still prevail. PMID- 12573758 TI - Poly(L-glutamic acid)-paclitaxel conjugate is a potent enhancer of tumor radiocurability. AB - PURPOSE: Conjugating drugs with polymeric carriers is one way to improve selective delivery to tumors. Poly (L-glutamic acid)-paclitaxel (PG-TXL) is one such conjugate. Compared with paclitaxel, its uptake, tumor retention, and antitumor efficacy are increased. Initial studies showed that PG-TXL given 24 h before or after radiotherapy enhanced tumor growth delay significantly more than paclitaxel. To determine if PG-TXL-induced enhancement is obtained in a more clinically relevant setting, we investigated PG-TXL effects on tumor cure. METHODS AND MATERIALS: Mice bearing 7-mm-diameter ovarian carcinomas were treated with PG-TXL at an equivalent paclitaxel dose of 80 mg/kg, single dose or 5 daily fractions of radiation or both PG-TXL and radiation. Treatment endpoint was TCD(50) (radiation dose yielding tumor control in 50% of mice). Acute radioresponse of jejunum, skin, and hair was determined for all treatments. RESULTS: PG-TXL dramatically improved tumor radioresponse, reducing TCD(50) of single-dose irradiation from 53.9 (52.2-55.5) Gy to 7.5 (4.5-10.7) Gy, an enhancement factor (EF) of 7.2. The drug improved the efficacy of fractionated irradiation even more, reducing the TCD(50) of 66.6 (62.8-90.4) Gy total fractionated dose to only 7.9 (4.3-11.5) Gy, for an EF of 8.4. PG-TXL did not affect normal tissue radioresponse resulting from either single or fractionated irradiation. CONCLUSION: PG-TXL dramatically potentiated tumor radiocurability after single-dose or fractionated irradiation without affecting acute normal tissue injury. To our knowledge, PG-TXL increased the therapeutic ratio of radiotherapy more than that previously reported for other taxanes, thus, PG-TXL has a high potential to improve clinical radiotherapy. PMID- 12573759 TI - EGFR blockade with ZD1839 ("Iressa") potentiates the antitumor effects of single and multiple fractions of ionizing radiation in human A431 squamous cell carcinoma. Epidermal growth factor receptor. AB - PURPOSE: Signaling pathways initiated by the epidermal growth factor receptor (EGFR) play important roles in the response to ionizing radiation. In this study the consequences of inhibiting the EGFR on the response of A431 cells (human vulvar squamous cell carcinoma cells that overexpress EGFR) to radiation, were investigated in vitro and in vivo, using the selective EGFR-tyrosine kinase inhibitor, ZD1839 ("Iressa"). METHODS AND MATERIALS: The effect of ZD1839 on proliferation, apoptosis, and clonogenic survival after radiation was determined in vitro. For in vivo studies, athymic nude mice with established subcutaneous A431 xenografts (approximately 100 mm(3)) were treated with either a single 10 Gy fraction or 4 daily 2.5 Gy fractions of radiation with or without ZD1839 (75 mg/kg/day intraperitoneally for 10 days) to determine effects on tumor growth delay. RESULTS: Treatment of A431 cells with ZD1839 in vitro reduced proliferation, increased apoptosis, and reduced clonogenic survival after radiation. Strikingly greater than additive effects of ZD1839 in combination with radiation on tumor growth delay were observed in vivo after either a single 10 Gy fraction (enhancement ratio: 1.5) or multiple 4 x 2.5 Gy fractions (enhancement ratio: 4). ZD1839 reduced tumor vascularity, as well as levels of vascular endothelial growth factor (VEGF) protein and mRNA induced by stimulation with epidermal growth factor (EGF), suggesting a possible role of inhibition of angiogenesis in the effect. CONCLUSIONS: Inhibiting EGFR-mediated signal transduction cascades with ZD1839 potentiates the antitumor effect of single and multiple fractions of radiation. These data provide preclinical rationale for clinical trials of EGFR inhibitors including ZD1839 in combination with radiation. PMID- 12573761 TI - Application of biological effective dose (BED) to estimate the duration of symptomatic relief and repopulation dose equivalent in palliative radiotherapy and chemotherapy. AB - PURPOSE: To investigate the potential for mathematic modeling in the assessment of symptom relief in palliative radiotherapy and cytotoxic chemotherapy. METHODS: The linear quadratic model of radiation effect with the overall treatment time and the daily dose equivalent of repopulation is modified to include the regrowth time after completion of therapy. RESULTS: The predicted times to restore the original tumor volumes after treatment are dependent on the biological effective dose (BED) delivered and the repopulation parameter (K); it is also possible to estimate K values from analysis of palliative treatment response durations. Hypofractionated radiotherapy given at a low total dose may produce long symptom relief in slow-growing tumors because of their low alpha/beta ratios (which confer high fraction sensitivity) and their slow regrowth rates. Cancers that have high alpha/beta ratios (which confer low fraction sensitivity), and that are expected to repopulate rapidly during therapy, are predicted to have short durations of symptom control. The BED concept can be used to estimate the equivalent dose of radiotherapy that will achieve the same duration of symptom relief as palliative chemotherapy. CONCLUSION: Relatively simple radiobiologic modeling can be used to guide decision-making regarding the choice of the most appropriate palliative schedules and has important implications in the design of radiotherapy or chemotherapy clinical trials. The methods described provide a rationalization for treatment selection in a wide variety of tumors. PMID- 12573760 TI - Comparing different NTCP models that predict the incidence of radiation pneumonitis. Normal tissue complication probability. AB - PURPOSE: To compare different normal tissue complication probability (NTCP) models to predict the incidence of radiation pneumonitis on the basis of the dose distribution in the lung. METHODS AND MATERIALS: The data from 382 breast cancer, malignant lymphoma, and inoperable non-small-cell lung cancer patients from two centers were studied. Radiation pneumonitis was scored using the Southwestern Oncology Group criteria. Dose-volume histograms of the lungs were calculated from the dose distributions that were corrected for dose per fraction effects. The dose-volume histogram of each patient was reduced to a single parameter using different local dose-effect relationships. Examples of single parameters were the mean lung dose (MLD) and the volume of lung receiving more than a threshold dose (V(Dth)). The parameters for the different NTCP models were fit to patient data using a maximum likelihood analysis. RESULTS: The best fit resulted in a linear local dose-effect relationship, with the MLD as the resulting single parameter. The relationship between the MLD and NTCP could be described with a median toxic dose (TD(50)) of 30.8 Gy and a steepness parameter m of 0.37. The best fit for the relationship between the V(Dth) and the NTCP was obtained with a D(th) of 13 Gy. The MLD model was found to be significantly better than the V(Dth) model (p <0.03). However, for 85% of the studied patients, the difference in NTCP calculated with both models was <10%, because of the high correlation between the two parameters. For dose distributions outside the range of the studied dose volume histograms, the difference in NTCP, using the two models could be >35%. For arbitrary dose distributions, an estimate of the uncertainty in the NTCP could be determined using the probability distribution of the parameter values of the Lyman-Kutcher-Burman model. CONCLUSION: The maximum likelihood method revealed that the underlying local dose-effect relation for radiation pneumonitis was linear (the MLD model), rather than a step function (the V(Dth) model). Thus, for the studied patient population, the MLD was the most accurate predictor for the incidence of radiation pneumonitis. PMID- 12573762 TI - Boron concentrations in brain during boron neutron capture therapy: in vivo measurements from the phase I trial EORTC 11961 using a gamma-ray telescope. AB - PURPOSE: Gamma-ray spectroscopic scans to measure boron concentrations in the irradiated volume were performed during treatment of 5 patients suffering from brain tumors with boron neutron capture therapy (BNCT). In BNCT, the dose that is meant to be targeted primarily to the tumor is the dose coming from the reaction 10B(n,alpha)7Li, which is determined by the boron concentration in tissue and the thermal neutron fluence rate. The boron distribution throughout the head of the patient during the treatment is therefore of major interest. The detection of the boron distribution during the irradiation was until now not possible. METHODS AND MATERIALS: Five patients suffering from glioblastoma multiforme and treated with BNCT in a dose escalation study were administered the boron compound, boron sulfhydryl (BSH; Na(2)B(12)H(11)SH). Boron concentrations were reconstructed from measurements performed with the gamma-ray telescope which detects locally the specific gamma rays produced by neutron capture in 10B and 1H. RESULTS: For all patients, at a 10B concentration in blood of 30 ppm, the boron concentration in nonoperated areas of the brain was very low, between 1 and 2.5 ppm. In the target volume, which included the area where the tumor had been removed and where remaining tumor cells have to be assumed, much higher boron concentrations were measured with large variations from one patient to another. Superficial tissue contained a higher concentration of 10B than the nonoperated areas of the brain, ranging between 8 and 15 ppm. CONCLUSIONS: The measured results correspond with previous tissue uptake studies, confirming that normal brain tissue hardly absorbs the boron compound BSH. Gamma-ray telescope measurements seem to be a promising method to provide information on the biodistribution of boron during therapy. Furthermore, it also opens the possibility of in vivo dosimetry. PMID- 12573763 TI - Optimization of conformal thoracic radiotherapy using cone-beam CT imaging for treatment verification. AB - PURPOSE: Megavoltage cone-beam computed tomography (MVCBCT) has been proposed for treatment verification in conformal radiotherapy. However, the doses required for such imaging may compromise the quality of the delivered dose distribution. The present paper explores the effect of cone-beam imaging on dose homogeneity and critical organ dose and the use of our new tool, adapted intensity-modulated radiation therapy (AIMRT). METHODS AND MATERIALS: Three types of treatment plans were devised (3D-CRT [three-dimensional conformal radiotherapy], IMRT [intensity modulated radiotherapy], and AIMRT) based on 4 patients with thoracic malignancies. MVCBCT fields were then integrated into the plans. The MVCBCT technique used 21 imaging portals at 10 degrees intervals. The MVCBCT apertures were shaped to conform to the planning target volume with a 6-mm margin. In a second set of plans, the field size was expanded by a further 2 cm. The unoptimized MVCBCT dose distribution was incorporated into the IMRT plan using AIMRT. RESULTS: Normal-tissue complication probability with MVCBCT is acceptable for all plans at the 66.6 Gy level, but exceeds tolerance for both 3D-CRT alone and 3D-CRT with MVCBCT at higher doses. In contrast, the use of AIMRT planning with MVCBCT allowed safe dose escalation to 85 Gy. Expanding the MVCBCT aperture provided better anatomic visibility with an acceptable lung dose. The results using IMRT with MVCBCT fell between the values measured for 3D-CRT and AIMRT with MVCBCT. CONCLUSION: The present study is the first to demonstrate that MVCBCT can be incorporated into 3D-CRT and IMRT planning with minimal effect on planning target volume homogeneity and dose to critical structures. This paves the way for highly conformal radiotherapy at greater doses delivered with increased confidence and safety. PMID- 12573764 TI - Intensity-modulated radiation therapy: a novel approach to the management of malignant pleural mesothelioma. AB - PURPOSE: Malignant pleural mesothelioma (MPM) causes symptoms and death mainly due to local progression, even after combined modality treatment. Poor local control after conventional radiotherapy may be due to the low dose of radiation that has been administered or to restriction of the target volume to avoid critical organs. Intensity-modulated radiation therapy (IMRT) has the potential to overcome these geometric/dosimetric constraints. METHODS AND MATERIALS: Seven patients with MPM who had an extrapleural pneumonectomy (EPP) were treated with adjuvant IMRT. The clinical target volume (CTV) included the surgically violated area inside the chest wall with particular attention to the insertion of the diaphragm, pleural reflections, and the deep margin of the thoracotomy incision. Treatment was delivered by intensity-modulated 6-MV photon beams using dynamic multileaf collimation. RESULTS: The CTV ranged from 2667 to 7286 mL. The average CTV covered to 50 Gy was 94% (range, 92% to 98%). Respiratory motion was minimal. The average volume of the boost areas covered by 60 Gy was 92% (range, 82% to 99%). Dose-volume constraints for normal tissue were met in almost all cases. Acute toxicity was mild to moderate. The most severe side effects were anorexia, nausea or vomiting, and dyspnea. Esophagitis was absent or mild. After a minimum of 13 months follow-up care there were no cases of disease recurrence within the ipsilateral hemithorax. CONCLUSION: Treatment of the extensive operative area after an EPP is feasible using IMRT. Input from the radiologist and from the surgeon in the planning process facilitates definition of the high dose volumes. In light of patients' tolerance to post-EPP IMRT, it may be feasible to incorporate systemic therapy, including novel biologic therapies into the treatment regimen. PMID- 12573765 TI - Intensity-modulated radiation therapy (IMRT) for locally advanced paranasal sinus tumors: incorporating clinical decisions in the optimization process. AB - PURPOSE: Intensity-modulated radiotherapy (IMRT) plans require decisions about priorities and tradeoffs among competing goals. This study evaluates the incorporation of various clinical decisions into the optimization system, using locally advanced paranasal sinus tumors as a model. METHODS AND MATERIALS: Thirteen patients with locally advanced paranasal sinus tumors were retrospectively replanned using inverse planning. Two clinical decisions were assumed: (1) Spare both optic pathways (OP), or (2) Spare only the contralateral OP. In each case, adequate tumor coverage (treated to 70 Gy in 35 fractions) was required. Two beamlet IMRT plans were thus developed for each patient using a class solution cost function. By altering one key variable at a time, different levels of risk of OP toxicity and planning target volume (PTV) compromise were compared in a systematic manner. The resulting clinical tradeoffs were analyzed using dosimetric criteria, tumor control probability (TCP), equivalent uniform dose (EUD), and normal tissue complication probability. RESULTS: Plan comparisons representing the two clinical decisions (sparing both OP and sparing only the contralateral OP), with respect to minimum dose, TCP, V(95), and EUD, demonstrated small, yet statistically significant, differences. However, when individual cases were analyzed further, significant PTV underdosage (>5%) was present in most cases for plans sparing both OP. In 6/13 cases (46%), PTV underdosage was between 5% and 15%, and in 3 cases (23%) was greater than 15%. By comparison, adequate PTV coverage was present in 8/13 cases (62%) for plans sparing only the contralateral OP. Mean target EUD comparisons between the two plans (including 9 cases where a clinical tradeoff between PTV coverage and OP sparing was required) were similar: 68.6 Gy and 69.1 Gy, respectively (p = 0.02). Mean TCP values for those 9 cases were 56.5 vs. 61.7, respectively (p = 0.006). CONCLUSIONS: In IMRT plans for paranasal sinus tumors, tradeoff values between OP toxicity and PTV coverage can be compared for different clinical decisions. The information derived can then be used to individualize the parameters within the optimization system. This process of determining clinical tradeoffs associated with different clinical decisions may be a useful tool in other sites. PMID- 12573766 TI - Potential role of intensity-modulated photons and protons in the treatment of the breast and regional nodes. AB - PURPOSE: To investigate, using comparative treatment planning, the potential improvements that could result through the use of intensity-modulated photons (intensity-modulated radiation therapy [IMRT]) and protons for the locoregional treatment of complex-target breast cancer. METHODS AND MATERIALS: Using CT data from a breast cancer patient, treatment plans were computed using "standard" photon/electron, IMRT, and forward-planned proton techniques. A dose of 50 Gy was prescribed to the target volume consisting of the involved breast, internal mammary, supraclavicular, and axillary nodes. The standard plan was designed using 6-MV X-ray beams to the breast, axillary, and supraclavicular areas and a mixture of 6-MV X-rays and 12-MeV electrons for the internal mammary nodes. Two IMRT (IMX1 and IMX2) plans were calculated for nine evenly spaced beams using dose-volume constraints to the organs at risk. For plan IMX1, precedence was given to optimizing the reduction in lung and heart dose while preserving target dose homogeneity. For plan IMX2, an increased precedence was given to the lungs, heart, and contralateral breast to further reduce doses to these organs and to study the effect on target coverage. The proton plan consisted of two oblique, energy-modulated fields. Target dose homogeneity and the doses to neighboring organs were both considered when comparing the different plans. RESULTS: For the standard plan, dose-volume histograms (DVHs) of the target volumes showed severe dose heterogeneity, whereas target coverage for the IMRT and proton plans was comparable. Lung DVHs for the standard and IMRT plans were also comparable, while the proton plan showed the best sparing over all dose levels. Mean doses to the ipsilateral lung for the three plans were found to be 17 Gy, 15 Gy, and 13 Gy for the standard, IMRT, and proton plans, respectively. For the heart, the IMRT plan delivered the highest mean dose (16 Gy), reflecting the extra dose delivered through this organ to spare the lungs. This was reduced somewhat by the standard plan (15 Gy), with the best sparing being provided by the proton plan (6 Gy). When the IMRT plan was reoptimized with an increased precedence to the normal tissues, the mean doses to all neighboring organs at risk could be reduced, but only at the cost of substantial target dose heterogeneity. CONCLUSIONS: In comparison with the standard plan, IMRT photons have the potential to greatly improve the target dose homogeneity with only a small increase in the doses delivered to the neighboring critical structures. However, when attempting to further reduce doses to the critical structures, substantial loss of target dose homogeneity was found. In conclusion, only the two-field, energy-modulated proton plan had the potential to preserve target dose homogeneity while simultaneously minimizing the dose delivered to both lungs, heart, and the contralateral breast. PMID- 12573767 TI - Megavoltage cone-beam computed tomography using a high-efficiency image receptor. AB - PURPOSE: To develop an image receptor capable of forming high-quality megavoltage CT images using modest radiation doses. METHODS AND MATERIALS: A flat-panel imaging system consisting of a conventional flat-panel sensor attached to a thick CsI scintillator has been fabricated. The scintillator consists of individual CsI crystals 8 mm thick by 0.38 mm x 0.38-mm pitch. Five sides of each crystal are coated with a reflecting powder/epoxy mixture, and the sixth side is in contact with the flat-panel sensor. A timing interface coordinates acquisition by the imaging system and pulsing of the linear accelerator. With this interface, as little as one accelerator pulse (0.023 cGy at the isocenter) can be used to form projection images. Different CT phantoms irradiated by a 6-MV X-ray beam have been imaged to evaluate the performance of the imaging system. The phantoms have been mounted on a rotating stage and rotated while 360 projection images are acquired in 48 s. These projections have been reconstructed using the Feldkamp cone-beam CT reconstruction algorithm. RESULTS AND DISCUSSION: Using an irradiation of 16 cGy (360 projections x 0.046 cGy/projection), the contrast resolution is approximately 1% for large objects. High-contrast structures as small as 1.2 mm are clearly visible. The reconstructed CT values are linear (R(2) = 0.98) for electron densities between 0.001 and 2.16 g/cm(3), and the reconstruction time for a 512 x 512 x 512 data set is 6 min. Images of an anthropomorphic phantom show that soft-tissue structures such as the heart, lung, kidneys, and liver are visible in the reconstructed images (16 cGy, 5-mm-thick slices). CONCLUSIONS: The acquisition of megavoltage CT images with soft-tissue contrast is possible with irradiations as small as 16 cGy. PMID- 12573768 TI - Procedures for high precision setup verification and correction of lung cancer patients using CT-simulation and digitally reconstructed radiographs (DRR). AB - PURPOSE: In a recent study, large systematic setup errors were detected in patients with lung cancer when a conventional simulation procedure was used to define and mark the treatment isocenter. In the present study, we describe a procedure to omit the session at a conventional simulator to remove simulation errors entirely. Isocenter definition and verification was performed at a computed tomography (CT) simulator, and digitally reconstructed radiographs (DRRs) were used for setup verification and correction at the treatment unit. METHODS AND MATERIALS: A CT simulation protocol was developed, in which radiopaque markers were used to verify the coincidence of the isocenter marked on the patients' skin with the isocenter defined in the planning CT scan. This protocol was evaluated for 20 patients. Subsequently, electronic portal images were acquired at the treatment unit. The three-dimensional setup error was established from a template match of the appropriate anatomy visible in two orthogonal beams with the corresponding anatomy in DRRs. An offline setup correction protocol was applied to reduce systematic setup errors. RESULTS: For all patients, the skin marks defined the planning CT scan isocenter to within +/- 1.5 mm in each of the three main directions. Random setup errors at the treatment unit were 1.8, 2.0, and 1.9 mm (1 SD) for the lateral (x), the superior-inferior (y), and the anterior-posterior (z) directions, respectively. With the use of the correction protocol, the systematic errors for x, y, and z were 1.5, 1.5, and 1.3 mm (1 SD). CONCLUSIONS: Because the distributions of treatment setup errors measured against DRRs obtained in our CT simulation were equal to previously obtained distributions measured against simulator films, conventional simulation can be omitted and DRRs are well-suited for setup verification. By adopting our CT simulation procedure, the large systematic simulation setup errors, which remain hidden if a conventional simulation is performed, can be avoided. PMID- 12573769 TI - Development of a semi-automatic alignment tool for accelerated localization of the prostate. AB - PURPOSE: Delivering high dose to prostate with external beam radiation has been shown to improve local tumor control. However, it has to be carefully performed to avoid partial target miss and delivering excessive dose to surrounding normal tissues. One way to achieve safe dose escalation is to precisely localize prostate immediately before daily treatment. Therefore, the radiation can be accurately delivered to the target. Once the prostate position is determined with high confidence, planning target volume (PTV) safety margin might be reduced for further reduction of rectal toxicity. A rapid computed tomography (CT)-based online prostate localization method is presented for this purpose. METHODS AND MATERIALS: Immediately before each treatment session, the patient is immobilized and undergoes a CT scan in the treatment position using a CT scanner situated in the treatment room. At the CT console, posterior, anterior, left, and right extents of the prostate are manually identified on each axial slice. The translational prostate displacements relative to the planned position are estimated by simultaneously fitting these identified extents from this CT scan to a template created from the finely sliced planning CT scan. A total of 106 serial CT scans from 8 prostate cancer patients were performed immediately before treatments and used to retrospectively evaluate the precision of this daily prostate targeting method. The three-dimensional displacement of the prostate with respect to its planned position was estimated. RESULTS: Five axial slices from each treatment CT scan were sufficient to produce a reliable correction when compared with prostate center of gravity (CoG) displacements calculated from physician-drawn contours. The differences (mean +/- SD) between these two correction schemes in the right-left (R/L), posterior-anterior (P/A), and superior-inferior (S/I) directions are 0.0 +/- 0.4 mm, 0.0 +/- 0.7 mm, and -0.4 +/- 1.9 mm, respectively. With daily CT extent-fitting correction, 97% of the scans showed that the entire posterior prostate gland was covered by PTV given a margin of 6 mm at the rectum-prostate interface and 10 mm elsewhere. In comparison, only 74% and 65% could be achieved by the corrections based on daily and weekly bony matching on portal images, respectively. CONCLUSIONS: Results show that daily CT extent fitting provides a precise correction of prostate position in terms of CoG. Identifying prostate extents on five axial CT slices at the CT console is less time-consuming compared with daily contouring of the prostate on many slices. Taking advantage of the prostate curvature in the longitudinal direction, this method also eliminates the necessity of identifying prostate base and apex. Therefore, it is clinically feasible and should provide an accelerated localization of the prostate immediately before daily treatment. PMID- 12573770 TI - Biologically effective dose (BED) for interstitial seed implants containing a mixture of radionuclides with different half-lives. AB - PURPOSE: To develop a tool for evaluating interstitial seed implants that contain a mixture of radionuclides with different half-lives and to demonstrate its utility by examining the clinical implications of prescribing to an isodose surface for such an implant. METHODS AND MATERIALS: A linear-quadratic model for continuous low dose rate irradiation was developed for permanent implants containing a mixture of radionuclides. Using a generalized equation for the biologically effective dose (BED), the effects of cell proliferation and sublethal damage repair were examined systematically for implants containing a mixture of radionuclides. A head-and-neck permanent seed implant that contained a mixture of (125)I and (103)Pd seeds was used to demonstrate the utility of the generalized BED. RESULTS: An equation of BED for implants containing a mixture of radionuclides with different half-lives was obtained. In such an implant, the effective cell kill was shown to depend strongly on the relative dose contributions from each radionuclide type; dose delivered by radionuclides with shorter half-life always resulted in more cell kill for any given sublethal damage repair and cell proliferation rates. Application of the BED formula to an implant containing a mixture of (125)I and (103)Pd seeds demonstrates that the conventional dose prescription to an isodose surface is not unique for such an implant. When the prescription dose was based on existing clinical experience of using (125)I seeds alone, mixing (103)Pd seeds with (125)I seeds would increase the cell kill. On the other hand, if the prescription dose were based on existing clinical experience of using (103)Pd seeds alone, mixing (125)I seeds with (103)Pd seeds in the same implant would create radiobiologically "cold" spots (i.e., an increase in cell survival) at locations where a major portion of the prescription dose is contributed by the (125)I seeds. For fast-growing tumors, these "cold" spots can become significant. CONCLUSIONS: Total dose alone is no longer sufficient for a complete characterization of a permanent seed implant containing a mixture of radionuclides with different half-lives due to the presence of cell proliferation and sublethal damage repair in the protracted dose delivery. BED provides a tool for evaluating the radiobiologic effects of mixing different type of radionuclides in the same implant. When radionuclides of different half-lives are mixed in a permanent implant, using the dose prescription established from existing clinical experience of implants with the longer half-life radionuclide would help to avoid radiobiologic "cold" spots. PMID- 12573771 TI - Influence of bladder and rectal volume on spatial variability of a bladder tumor during radical radiotherapy. AB - PURPOSE: To assess the spatial variability of a bladder tumor relative to the planning target volume boundaries during radical radiotherapy, and furthermore to develop strategies to reduce spatial variability. METHODS AND MATERIALS: Seventeen patients with solitary T2-T4N0M0 bladder cancer were treated with a technique delivering 40 Gy/2 Gy in 20 fractions to the whole bladder with a concomitant boost to the bladder tumor of 20 Gy in 1 Gy fractions in an overall time of 4 weeks. CT scans were made weekly, immediately after treatment, and matched with the planning CT scan. Spatial variability of the tumor, as well as bladder volume and rectal diameter, were scored for each patient each week. RESULTS: In 65% of patients, a part of the tumor appeared outside the planning target volume boundaries at least one time during the course of radiotherapy. No consistent relation of this variability with time was found. Bladder volumes and rectal diameters showed marked variability during the course of treatment. A large initial bladder volume and rectal diameter predicted a large volume variation and a large tumor spatial variability. CONCLUSION: In this study, a margin of 1.5 to 2 cm seemed to be inadequate in 65% of the patients with respect to spatial variability. Bladder volume and rectal diameter were found to be predictive for spatial variability of a bladder tumor during concomitant boost radiotherapy. PMID- 12573772 TI - Dose effects of stents in intravascular brachytherapy for in-stent restenosis: a Monte Carlo calculation. AB - PURPOSE: Intravascular brachytherapy (IVBT) has been recognized as a preferred treatment for coronary in-stent restenosis (ISR) in routine practice. Stents made of high-Z materials will inevitably perturb the dose distribution of IVBT. In this work, we have conducted a systematic study on these dose perturbations for three commercially available IVBT sources. METHODS AND MATERIALS: The EGSnrc Monte Carlo codes were used to calculate the dose distributions for the 90Sr, 32P, and 192Ir IVBT sources with and without a metallic stent in place. The ring stent type made of different material and with different strut size, metallic surface area (MSA), and radius was studied. RESULTS: Calculations show that dose enhancement of 5% to 20% occurs inside stent in the region close to the stent struts (luminal side) for all three sources. In the region outside stent (adventitial side), dose reduction of 5% to 20% is observed for a beta source, whereas the dose effect is negligible for the gamma source. For a given stent design, the tantalum stent yields a larger dose effect than other stents made of steel, Ti, Ni, or nitinol. It is found that the dose effect significantly depends on strut thickness, and it is strongly correlated to MSA. The MSA may be used to characterize the dose effect of a stent. Sample empiric equations to relate the dose perturbations to MSA for a given source, a stent material, and a strut thickness were derived. CONCLUSIONS: The dose perturbations due to the presence of metallic stents were found to be significant in IVBT for ISR. The dose effects of a stent can be estimated from its MSA based on derived empiric equations. The data presented are practically useful to consider the dose effects of stents in dose evaluation/treatment planning for using IVBT to treat ISR. PMID- 12573773 TI - Glioma treatment, radionecrosis, and growth factors. In regard to Levin et al., IJROBP 2002;53:58-66. PMID- 12573775 TI - Palliation of esophageal cancer: there is no clear winner. PMID- 12573777 TI - Evaluating locoregional recurrence in breast cancer. PMID- 12573779 TI - Commentary on IMRT and cancer of the prostate. Intensity-modulated radiation therapy. PMID- 12573781 TI - In regard to Huang et al.: Intensity-modulated radiation therapy for pediatric medulloblastoma: early report on the reduction of ototoxicity. IJROBP 2002;52:599 605. PMID- 12573783 TI - The role of ultrasound in biliary and pancreatic diseases. AB - The possibilities and the limits of transabdominal ultrasonography (US) in the diagnosis of bilio-pancreatic diseases are reviewed here in the light of the last 10 years' research. US remains the method of choice for the diagnosis of gallstones and is generally accepted as an initial imaging technique in gallstone complications, such as acute cholecystitis. Moreover the method can be useful for the detection of the biliary complications after laparoscopic cholecystectomy and after liver transplantation. US is still considered the first diagnostic procedure when stones are suspected in the common bile duct. The use of color Doppler can provide a differential diagnosis of gallbladder cancer with respect to other benign inflammatory or polypoid lesions. Color Doppler US allows to detect vascular complications of acute pancreatitis such as pseudoaneurysms. US is still considered useful for the initial screening of the pancreatic cancer. However, for staging other imaging techniques must be employed. With US useful informations are obtained in the diagnosis of cystic tumors of the pancreas and of pancreatic metastases. US is generally of little use for the diagnosis of endocrine tumors. PMID- 12573784 TI - Ultrasonographic analysis of arteriosclerotic plaques in the internal carotid artery. AB - OBJECTIVES: The aim of this study was to investigate whether ultrasonography can characterise plaque morphology and surfaces independent of the observer. METHODS: Computer-assisted image analysis of the grey scales of B-mode scans from 15 patients with stenoses of the internal carotid artery was performed and compared with the histopathological reports. In vitro angioscopy, was used to visualise the plaque surfaces of the thromboendarterectomy specimens. RESULT: Assessment of the internal plaque structure by ultrasound showed close agreement between the two observers (P<0.01) without correlation with the histopathological results. Ultrasonography was able to characterise the plaque surfaces in 93% of cases. CONCLUSION: This investigation showed that standardised computer-assisted analysis of the internal plaque structure correlates less closely than histological investigation, but agrees closely with the surface structure of the plaque. The exclusive use of digital image processing and standardisation of the investigative technique are expected to provide even better results. PMID- 12573785 TI - Control system analysis of visually evoked blood flow regulation in humans under normocapnia and hypercapnia. AB - OBJECTIVE: Among other factors, the cerebral blood flow (CBF) is regulated in accordance to the arterial CO(2) tension and the cortical activity. The CO(2) test is commonly used to measure the vascular reserve capacity. Most functional imaging studies rely on the activity-flow coupling (AFC). We aimed to combine both challenges in order to increase the insight into mechanisms of CBF regulation. METHODS: Fifteen healthy students underwent a functional transcranial Doppler test using a visual stimulation paradigm: firstly under normocapnia and secondly under conditions of hypercapnia. Hypercapnia was induced by breathing a carbogene gas mixture of 5% CO(2) and 95% O(2). The entire time course of flow velocity adaptation in the posterior cerebral artery (PCA) was analyzed mathematically using a control system approach. RESULTS: Resting CBF velocities increased by nearly 26% under conditions of hypercapnia, whereas the slight increase in arterial blood pressure (ABP) and the decrease in the Pourcelot Pulsatility index (PI) were statistically not significant. From the control system parameters which were time delay, rate time, gain, attenuation and natural frequency, only the parameter rate time, indicative for the initial steepness of flow velocity increase, showed a statistically significant decrease, consistently for the peak systolic and enddiastolic flow velocity data. As concluded from the unchanged gain parameter the absolute amount of blood flow evoked by the same visual stimulus increased also by 26%. CONCLUSION: Evaluated by Doppler measurements hypercapnia seems to influence the AFC in two ways: It decreases the steepness of the initial increase in blood flow velocity and enhances the absolute amount of blood flow evoked by the same stimulus. PMID- 12573786 TI - Effect of hypertension and candesartan on the blood flow velocity of the extraocular vessels in hypertensive patients. AB - OBJECTIVE: To define alterations in the blood flow velocities of the ophthalmic artery (OA), central retinal artery (CRA), posterior ciliary artery (PCA) in essential hypertension and to evaluate the effect of a new antihypertensive drug, candesartan which is an angiotensin II receptor antagonist, on the blood flow velocity in hypertensive patients. METHODS: Blood flow velocity and resistive index (RI) of the OA, CRA, and PCA were measured in 22 hypertensive patients off medication and 15 controls by color Doppler imaging. After treatment with candesartan, blood flow velocity and RI were again measured in the hypertensive patients. RESULTS: In controls (n=15), the OA had a mean peak systolic flow velocity (PSFV) of 48.1+/-2.6 cm/s, mean end diastolic flow velocity (EDFV) of 16+/-1.0 cm/s, and RI of 0.65+/-0.01; the CRA had a PSFV of 20.8+/-0.4 cm/s, EDFV of 9.4+/-0.3 cm/s, and RI of 0.54+/-0.01; the PCA had a PSFV of 23.6+/-0.7 cm/s, EDFV of 11.2+/-0.3 cm/s, and RI of 0.52+/-0.01. There was a significant decrease in the PSFV and EDFV of the vessels in the medication free hypertensive patients when compared with controls (P<0.05). In the hypertensive patients off medication (n=22), the OA had a PSFV of 29.4+/-1.2 cm/s, EDFV of 10.4+/-0.5 cm/s, and RI of 0.71+/-0.01; the CRA had a PSFV of 15.1+/-0.6 cm/s, EDFV of 5.4+/-0.3 cm/s, and RI of 0.65+/-0.02; the PCA had a PSFV of 17.2+/-0.6 cm/s, EDFV of 6.7+/-0.3 cm/s, and RI of 0.61+/-0.01. RI measured in the OA, CRA, PCA were significantly increased in the hypertensive patients when compared with the controls (P<0.05). In hypertensive patients after medication (n=22), OA had a PSFV of 38.3+/-2.5 cm/s, EDFV of 12.3+/-0.7 cm/s, and RI of 0.68+/-0.01; CRA had a PSFV of 19.2+/ 0.5 cm/s, EDFV of 7.8+/-0.3 cm/s, and RI of 0.59+/-0.01; PCA had a PSFV of 20.8+/ 0.8 cm/s, EDFV of 9.2+/-0.4 cm/s, and RI of 0.56+/-0.01. There was a significant increase in the blood flow velocities of the OA, CRA, PCA (P<0.05) and significant decrease in the RI values in the treated hypertensive patients when compared with the controls (P<0.05). But blood flow velocities and RI values did not reach the control level. CONCLUSION: The increase in the RI values and the decrease in the blood flow velocity of extraocular vessels in the hypertensive patients are thought to be caused by increased peripheral resistance in the vessels of the eye and orbit. Although, it increases blood flow velocity and decreases RI significantly, candesartan treatment in the hypertensive patients cannot increase blood flow velocity and decrease RI to the control level. PMID- 12573787 TI - Color Doppler sonography in hypothyroidism. AB - BACKGROUND: The value of color Doppler sonography in thyroid disease continues to be a matter of debate. Over the past few years, several studies have proved unable to yield unequivocal results. Only a few studies concerning color Doppler sonography in patients with hypothyroidism have been published. METHODS: 89 patients with hypothyroidism have been evaluated. They were examined clinically; laboratory tests on thyroid function and color Doppler sonography have been performed. The color flow distribution and intensity were estimated and the fastest flow velocity (PSV) detectable with a pw-doppler was registered. The color pattern was graded from 0 to III as has been described by others and the color Doppler findings were then correlated to both the clinical picture and the laboratory results. RESULTS: 56 of the 89 hypothyroid patients showed pattern 0 with a PSV of 22 cm/s. In 33 patients different degrees of increased parenchymal color could be found with a concordant PSV: 16 patients showed pattern I with a PSV of 39 cm/s; 11 showed pattern II with PSV 58 cm/s, and 6 showed pattern III with PSV 63 cm/s. Regarding the corresponding clinical and laboratory variables, there was a very close correlation between color intensity and anti-Tg/anti-TPO antibody levels: pattern 0: anti-Tg 474 IU/ml, anti-TPO-Ab 810 IU/ml; pattern I: 1053/1733; pattern II: 1774/2432; pattern III: 1951/2633. Some correlation could also be found for the TSH values and the calculated volume of the thyroid gland, whereas the duration of hypothyroidism showed an inverse correlation to color intensity. (Pattern 0: TSH 3.1 mE/ml, volume 9.2 ml, duration 43 months; pattern I: 4.2 mE/l, 15.7 ml, 24 mos.; pattern II: 11.5 mE/l, 22.3 ml, 16 mos.; pattern III: 38.2 mE/l, 34.3 ml, 10 mos, respectively). CONCLUSIONS: The color Doppler pattern of intense hypervascularization of the thyroid gland formerly attributed only to the hyperthyroid state of active Graves' Disease can also be seen in hypothyroidism. Our data support the concept that the color flow appearance is not the result of stimulated thyroid hormone production, but a measure of the activity of an autoimmune process. PMID- 12573788 TI - Diagnostic value of high-resolution B-mode and power-mode sonography in the follow-up of thyroid cancer. AB - OBJECTIVE: Ultrasonography is an established diagnostic modality in the follow-up of thyroid cancer. Color flow Doppler has been proposed by some authors as an additional tool for differentiating benign from malignant cervical lesions in various types of head and neck cancer. Over the last few years, a new generation of high-resolution ultrasound platforms with the "power-mode" feature has become available, that also enables the imaging of small vessel blood flow. The objective of our study was to find ways of optimizing the differentiation of benign and malignant cervical tumors in thyroid cancer follow-up by means of sonography. METHODS: Hundred and twelve cervical lesions in 90 patients with thyroid cancer were evaluated by high-end ultrasonography (Sonoline Elegra, Siemens) using a small-part transducer (7.5 L 40, Siemens). B-mode sonography was performed at a frequency of 8 MHz. The Solbiati index (SI= ratio of largest to smallest diameter), configuration, echogenicity, intranodular structures, and margins were assessed. Perinodular and intranodular blood flow was evaluated by color flow Doppler (PRF 1250 Hz for conventional color flow Doppler, 868 Hz for power-mode Doppler). Possible malignancy was validated by histology, cytology, scintigraphy, and follow-up. Thirty five lesions were benign (diameter 0.4-3.0 cm) and 77 were malignant (0.4-5.4 cm). The patients were randomized into a test group and a learning group to determine the diagnostic value of various ultrasound criteria by means of statistical analysis. In the learning group, decision rules based on the dichotomized criteria were developed using a logistic regression model. Sensitivity and specificity of these decision rules were then evaluated in the test group. RESULTS: The presence of an echocomplex pattern or irregular hyperechoic small intranodular structures (criterion A) and the presence of an irregular diffuse intranodular blood flow (criterion B) are the best indicators of malignancy, whereas an SI >>2 is highly indicative of benign changes. Color flow Doppler is a useful addition to B-mode scanning for distinguishing benign and malignant neoplasms in the follow-up of thyroid cancer. Power-mode Doppler sonography significantly improves imaging of perinodular and intranodular blood flow when compared with conventional color flow Doppler. CONCLUSION: We propose the following decision rules based on a combination of the criteria above: (A) and (B) fulfilled: malignant, if SI< or =4; (B) but not (A) fulfilled: malignant, if SI< or =3; (A) but not (B) fulfilled: malignant, if SI< or =2; neither (A) nor (B) fulfilled: malignant, if SI approximately equal to 1 (sensitivity: 90%; specificity: 82%; accuracy 88%). PMID- 12573789 TI - Reducing inter-observer variation in thyroid volume calculation using a new formula and technique. AB - OBJECTIVE: Ultrasound (US) measurement of thyroid volume is used in diagnosis and follow-up of patients with thyroid disease. We assessed a new formula and technique for thyroid volume calculation, based on an automatically continuous trace transverse surface area (aTSA) calculation, to ascertain where this technique could reduce inter-observer variation. METHODS: Three observers with a different level of US expertise, using a 12-5 MHz linear transducer, examined 25 volunteers. Inter-observer variations were calculated for diameter measurements and for the thyroid lobe volume calculations using the single factor ANOVA method. RESULTS: Using the new technique and formula, no statistically significant differences existed, in contradistinction to using the classical formula for an ellipsoid (P=0.02). CONCLUSION: When thyroid volume measurements are required, we recommend using a new method of thyroid volume calculation based on the use of an automatically calculated continuous trace aTSA measurement to avoid significant inter-observer variation in calculation of thyroid lobe volumes. PMID- 12573790 TI - The value of intraoperative ultrasound during hepatic resection compared with improved preoperative magnetic resonance imaging. AB - BACKGROUND: Patients with colorectal metastases confined to the liver may be cured by resection. Combined bimanual palpation and intraoperative ultrasound (IOUS) augment the detection of colorectal hepatic metastases. The importance of IOUS in the surgical management of hepatic tumors has been demonstrated and should arguably be considered the final diagnostic procedure. OBJECTIVE: To determine the relevance of routine IOUS prior to hepatic resection compared with improved preoperative Magnetic resonance imaging (MRI). PATIENTS AND METHODS: Eighty patients with metastatic colorectal adenocarcinoma underwent hepatic resection between 1998 and 2001. The IOUS results were compared with preoperative MRI, bimanual palpation, and resection histopathology. The preoperative surgical plan was compared with the surgical procedure performed. Data were retrospectively analyzed. RESULTS: IOUS provided additional useful information not available preoperatively for 37 (47%) patients, including the identification of subcentimetre metastatic lesions, characterization of the lesion, and the anatomy of the hepatic vasculature. The preoperative surgical plan was changed secondary to the IOUS findings alone in 14/80 (18%) patients. IOUS did not provide any additional useful information for 43 (53%) patients. A correlation was demonstrated between the preoperative diagnosis, intraoperative findings, and resection histopathology. CONCLUSION: Accurate diagnostic studies facilitate critically decisive actions during planned hepatic resection(s). The current findings suggest that IOUS provide the most useful additional information for hepatic lesions, despite recent improvements in preoperative MRI scanning. Furthermore, routine IOUS should be employed during hepatic resection for colorectal metastases. PMID- 12573791 TI - Sonographic diagnosis of hydatidosis: the sign of the cyst wall. AB - The Authors evaluate the diagnostic accuracy of echography in hepatic echinococcosis, particularly in the completely liquid types of cysts without septa or hydatid sand and in the solidified forms where the differential diagnosis with other hepatic pathologies can be difficult. The study was carried out with a 7.5 MHz probe and the results were compared with those from the use of a 3.5 MHz probe and from CT, serology for hydatidosis and echoguided sampling. Seventy one patients, 36 with liquid hepatic lesions and 35 with solid hepatic lesions (31 hydatid and 40 non-parasitic), were evaluated echographically, first with a 3.5 MHz probe and subsequently with a 7.5 MHz probe. All the patients underwent serology for hydatidosis. All the liquid lesions were subjected to percutaneous echoguided aspiration and all the solid lesions to CT; echoguided biopsy was performed in only 24 of the latter lesions. With the 7.5 MHz probe, a wall was revealed in 96.77% of parasitic cysts but in only 5% of non-hydatid ones. Moreover, using the 7.5 MHz probe, it was possible to see a triple-layered aspect of the wall in the hydatid cysts. The results obtained with the 7.5 MHz probe demonstrate a high diagnostic accuracy comparable to that of MR. PMID- 12573792 TI - The diagnostic value of levovist in Doppler imaging of visceral arteries in patients with abdominal angina before and after angioplasty. AB - PURPOSE: To assess the diagnostic value of Levovist in the ultrasound imaging of visceral arteries in patients with clinical symptoms of abdominal angina, before and after percutaneous transluminal angioplasty (PTA). MATERIAL AND METHOD: During a 12-month period (2000/2001) five patients with visceral arterial stenoses had ultrasound examinations and a subsequent PTA procedure. Indications for ultrasound examination were abdominal angina symptoms persisting for 3-5 years, (postprandial abdominal pain, diarrhea, and vomiting). In all patients ultrasound examinations were performed using color and spectral Doppler before and after Levovist injections. Color Doppler images and maximum blood flow velocity in stenosed visceral arteries were assessed. Patients underwent control Doppler examinations with Levovist injections to assess the effect of PTA. RESULTS: In three patients conventional Doppler examination did not allow proper evaluation of visceral arteries, because of weak color and spectral Doppler signal and in two remaining patients visceral arteries were not visualized at all. In all five patients strong enhancement of color and spectral Doppler signal was observed after Levovist administration. In all these cases a hemodynamically significant stenosis was diagnosed: coeliac trunk-2 and superior mesenteric artery-3. PTA was performed successfully in these patients. In one of them ultrasound examination done before Levovist injection allowed good visualization of treated SMA and showed good PTA result. In the remaining four patients Doppler examination with the use of Levovist demonstrated visceral arteries well and confirmed successful PTA procedures. CONCLUSIONS: The use of Levovist makes the diagnostic efficiency of Doppler examinations much higher. In most cases it allows an unequivocal diagnosis of visceral artery stenosis in patients with abdominal angina symptoms. The Doppler examination with the use of Levovist is the method of choice in follow-up after PTA. PMID- 12573793 TI - The gender of the examiner, the state of the pregnancy and women's perception of transvaginal sonography in the first trimester. AB - OBJECTIVES: The objectives are to study the correlation between the women's perception of transvaginal sonography (TVS) and the gender of the examiner and the state of the pregnancy. METHODS: The population studied was unselected women attending the early assessment unit at Sharoe Green Hospital with various complications of early pregnancy. All the women were in the first trimester. Only women who had TVS and who gave informed consent were included. RESULTS: Four hundred and twenty five (425) women were recruited. A male doctor examined 215 (50.6%), while a female doctor examined the remaining 210 (49.4%). Two hundred and ninety eight (70.1%) of the pregnancies were viable and 127 (29.9%) were non viable. About 98.1% of the women found TVS acceptable. There was no statistically significant difference in the women's perception of the procedure with regard to the gender of the examiner or the state of the pregnancy. CONCLUSIONS: The majority of women (98.1%) perceived TVS favourably. The gender of the examiner and the state of the pregnancy have no influence on the women's perception of the procedure. PMID- 12573794 TI - Heating caused by selected pulsed Doppler and physiotherapy ultrasound beams measured using thermal test objects. AB - OBJECTIVE: To assess the heating caused by selected pulsed Doppler and physiotherapy ultrasound beams by measurements made using thermal test objects. METHOD: Thermal test objects were used to measure temperature rises in selected ultrasound fields. These were compared with theoretical predictions based on standard exposure measurements. A separate thermocouple was used to measure heating at the transducer surface. RESULTS: Temperature rises of up to 6 degrees C were measured for Doppler fields using a thermal test object. The attenuation corrected temperature rises that were measured generally agreed with calculated Thermal Indices. Temperature rises of up to 2 degrees C were observed for physiotherapy ultrasound fields in pulsed operation. CONCLUSION: Significant overlap between the measured temperature rises of selected pulsed Doppler and physiotherapy ultrasound fields was observed. PMID- 12573795 TI - Incoherent imaging using continuous wave ultrasound. A preliminary study using bovine intervertebral discs. AB - As an object rotates with respect to a stationary planar ultrasonic beam, the scattering centres within the object return echoes that are Doppler-shifted in frequency by amounts depending on the velocities of the individual scatterers. The backscattered echo amplitude at any particular frequency is the line integral of the scattered radiation at the cross-range corresponding to that frequency. The amplitude as a function of frequency can be interpreted as a tomographic projection. A tomographic reconstruction algorithm can produce an image of the distribution of scattering centres in the insonified object from these projections. This paper describes the development and characterisation of a microscanner to investigate the approach of using continuous wave ultrasound for three-dimensional cross-sectional imaging. The results of preliminary tissue investigation, conducted using bovine coccygeal intervertebral discs, are described. The radial imaging resolution improves as the Doppler frequency resolution improves but the circumferential resolution degrades proportionally. As the number of projections increases, there is a finite increase in image quality. Two- and three-dimensional images of the intervertebral disc reveal an alternate light and dark banding pattern that is characteristic of the laminar structure of the annulus fibrosus. PMID- 12573796 TI - An unusual presentation of neonatal adrenal hemorrhage. AB - Case report of a neonate presenting within 1 h of birth with right side scrotal swelling and a dusky skin discoloration thought clinically to be a hydrocele or testicular torsion. Ultrasound examination demonstrated bilateral hydroceles, more prominent on the right, scrotal oedema and a perinephric loculated fluid collection secondary to right adrenal hemorrhage. We review the ultrasound appearance of the normal and haemorrhagic neonatal adrenal and discuss the differential diagnosis of both neonatal adrenal lesions and scrotal swellings. The role and benefits of ultrasound in the primary diagnosis and management of neonatal adrenal hemorrhage is emphasized. PMID- 12573798 TI - VIP and PACAP display different vasodilatory effects in rabbit coronary and cerebral arteries. AB - Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase activating polypeptide (PACAP) are closely related peptides with wide distribution in the nervous system. The aim of the present study was to investigate functional characteristics and the influence of sex steroids on the vasodilatory effects of these two peptides in cerebral and coronary vessels from female New Zealand White (NZW) rabbits. The localization and concentration of VIP and PACAP in cardiovascular tissue was evaluated using immunohistochemistry and radioimmunoassays. The vasodilatory effects of VIP and PACAP were investigated using myographs, allowing isometric tension recordings. In order to evaluate the influence of steroid hormones, the rabbits were ovariectomized and randomized to treatment for 4 weeks with 17beta-estradiol (E(2)), Norethindrone Acetate (NETA), E(2)+NETA or placebo. Ring segments of the posterior cerebral artery, the right proximal coronary artery and the distal left coronary artery were examined. The highest concentrations of VIP/PACAP were observed in cerebral and coronary arteries: 5.0/5.7 and 2.8/3.5 pmol/g, respectively. The peptides were localized in nerve fibres innervating the arteries. Both peptides produced dose-dependent vasodilatory responses in all vessels investigated. While the effects of PACAP were identical in cerebral and coronary arterial segments, the effects of VIP displayed significant differences (E(max), pI(2), Hill-slope). Treatment with sex steroids induced no changes in the vascular effects of the two peptides. These results indicate different mechanisms of action for the vasodilating effects of the two closely related peptides VIP and PACAP in different areas of the coronary and cerebrovascular tree. Treatment with female sex steroids does not seem to change these mechanisms. PMID- 12573799 TI - Adrenomedullin stimulates somatostatin and thus inhibits histamine and acid secretion in the fundus of the stomach. AB - Adrenomedullin has recently been localized to enterochromaffin-like (ECL) and chief cells in the gastric fundus. It has been proposed that adrenomedullin may play a role in gastric mucosal defense and repair. In the present study, we have used the isolated, luminally perfused mouse stomach and superfused rat fundic segments to examine the effect of adrenomedullin on exocrine and endocrine secretion in this region of the stomach. Addition of adrenomedullin (1 pM to 1 microM) to the isolated mouse stomach caused a concentration-dependent decrease in acid secretion. The EC(50) value was 1.4 x 10(-9) and maximal inhibition of acid secretion was obtained at a concentration of 1 microM (31+/-4% below basal level, P<0.001). In rat fundic segments, superfusion with adrenomedullin (0.1 pM to 0.1 microM) caused a concentration-dependent increase in somatostatin secretion (EC(50), 1 x 10(-10)) that was accompanied by a reciprocal decrease in histamine secretion (EC(50), 1.2 x 10(-11)). Maximal stimulation of somatostatin secretion (60+/-5% above basal level, P<0.001) and inhibition of histamine secretion (50+/-5% below basal level, P<0.01) was obtained at a concentration of 0.1 microM. Changes in acid and histamine secretion induced by adrenomedullin reflected changes in somatostatin secretion and could be abolished by addition of somatostatin antibody. The axonal blocker, tetrodotoxin, also abolished the somatostatin and, consequently, the acid and histamine responses to adrenomedullin, implying that the effect of adrenomedullin on somatostatin secretion was mediated via activation of intramural neurons. We conclude that adrenomedullin, acting via intramural fundic neurons, stimulates somatostatin and thus inhibits histamine and acid secretion. This represents one mechanism by which adrenomedullin might enhance mucosal defense and repair. PMID- 12573800 TI - Prohormone convertase 2 enzymatic activity and its regulation in neuro-endocrine cells and tissues. AB - We used the fluorometric substrate, pGlu-Arg-Thr-Lys-Arg-MCA and the C-terminal peptide of human 7B2(155-185), a specific inhibitor of prohormone convertase 2 (PC2), to specifically measure the enzymatic activity of the prohormone convertases, PC2. Using lysates from the pancreatic alpha cell line, alphaTC1-6 cells, which contain moderate levels of PC2 enzymatic activity, we determined that the PC2 assay was linear with respect to time of incubation and protein added and had a pH optimum of 5.5 and a calcium optimum of 2.5 mM. Rat pituitary contained high levels of PC2 enzymatic activity, while the hypothalamus and other brain regions contained moderate levels. This enzyme assay was used to document that both mice null for PC2 as well as mice null for the PC2 cofactor, 7B2, had only trace levels of PC2 activity in various brain regions, while mice heterozygous for these alleles had approximately half of the PC2 activity in most brain regions. PC2 enzymatic activity and PC2 mRNA levels were somewhat discordant suggesting that PC2 mRNA levels do not always reflect PC2 enzymatic activity. PMID- 12573801 TI - Bradykinin counteracts the stimulatory effect of angiotensin-(1-7) on the proximal tubule Na+ -ATPase activity through B2 receptor. AB - Recently, we demonstrated that angiotensin-(1-7) (Ang-(1-7)) stimulates the Na(+) ATPase activity through a losartan-sensitive angiotensin receptor, whereas bradykinin inhibits the enzyme activity through the B(2) receptor [Regul. Pept. 91 (2000) 45; Pharmacol. Rev. 32 (1980) 1]. In the present paper, the effect of bradykinin (BK) on Ang-(1-7)-stimulated Na(+)-ATPase activity was evaluated. Preincubation of Na(+)-ATPase with 10(-9) M Ang-(1-7) increases enzyme activity from 7.9+/-0.9 to 14.1+/-1.5 nmol Pi mg(-1) min(-1), corresponding to an increase of 79% (p<0.05). This effect is reverted by bradykinin in a dose-dependent manner (10(-14)-10(-8) M), reaching maximal inhibitory effect at 10(-9) M. Des-Arg(9) bradykinin (DABK), an agonist of B(1) receptor, at the concentrations of 10(-9) 10(-7) M, does not mimic the BK inhibitory effect, and des-Arg(9)-[Leu(8)]-BK (DALBK), a B(1) receptor antagonist, at the concentrations of 10(-10)-10(-7) M, does not prevent the inhibitory effect of BK on Ang-(1-7)-stimulated enzyme. On the other hand, HOE 140, an antagonist of B(2) receptor, abolishes the inhibitory effect of BK on the Ang-(1-7)-stimulated enzyme in a dose-dependent manner, reaching maximal effect at 10(-7) M. Taken together, these data indicate that stimulation of B(2) receptors by BK can counteract the stimulatory effect of Ang (1-7) on the proximal tubule Na(+)-ATPase activity. PMID- 12573802 TI - Expression of mRNA for PACAP and its receptors in intra- and extra-adrenal human pheochromocytomas and their relationship to catecholamine synthesis. AB - PURPOSE: Pituitary adenylate cyclase-activating polypeptide (PACAP), a member of the secretin/glucagons/vasoactive intestinal peptide family, induces the expression of catecholamine-synthesizing enzymes in adrenal medullary cells. In addition, PACAP and its receptor have been detected in human pheochromocytoma tissues, though it is not yet known whether PACAP enhances the expression of genes encoding catecholamine-synthesizing enzymes. To address this question, we analyzed PACAP, PACAP receptor, and tyrosine hydroxylase (TH) and phenylethanolamine-N-methyltransferase (PNMT) mRNAs in pheochromocytomas. METHODS: The levels of the mRNA for PACAP and vasoactive intestinal peptide (VIP), and their receptors, and for TH and PNMT were measured by RT-PCR or real time PCR analysis, and the concentrations of catecholamines were measured by HPLC in 24 intra-adrenal and six extra-adrenal pheochromocytomas. RESULTS: mRNA expression of PACAP and its receptor VPAC1R were detected in many pheochromocytomas (24/30 and 29/30, respectively), but mRNA expression of the PAC1R and VPAC2R receptor subtypes were detected in only one of six extra-adrenal pheochromocytomas. PACAP mRNA expression correlated with TH (p=0.0018) and PNMT (p=0.05) mRNA expression, as well as epinephrine (p=0.0342) levels in 16 intra adrenal pheochromocytomas. CONCLUSION: Our findings support a possible role for PACAP in the regulation of expression of genes encoding catecholamine synthesizing enzymes in intra-adrenal pheochromocytomas. PMID- 12573803 TI - Manipulation of neonatal gonadal steroid milieu and leptin secretion in later life in male and female rats. AB - The mechanism underlying the gender-based difference in circulating leptin levels (females>males) is still uncertain, because the difference persists even after adjustment for fat mass and sex steroid concentrations. In this study, we tested the possibility that the neonatal sex steroid milieu, which is critical for the sexual differentiation of the brain, may permanently affect leptin secretion in rats of both sexes. Male rats were neonatally castrated (NC), and females were neonatally androgenized (NA) by testosterone (T). Two subsets of the NC males were given T on postnatal day 1 or 29. Appropriate controls for all these groups were prepared. The animals were sacrificed on postnatal day 57, and at this age, the percent body fat was similar among all the male and female groups. NC males had a two-fold, significantly higher level of leptin than intact males. This hyperleptinemia induced by NC was prevented by T when it was given neonatally, but not on the day 29. By contrast, NA for females was without effect on leptin titers in later life. These results suggest that neonatal T in male rats may, at least in part, mediate the sex-related difference in leptin secretion that becomes apparent in later life. However, as intact females still had significantly higher leptin titers than NC males, it is very likely that additional factors may also be responsible for the sexually dimorphic leptin secretion in rats. PMID- 12573804 TI - Effects of morphine administration and its withdrawal on rat brain aminopeptidase activities. AB - The endogenous opioid neuropeptide system seems to be involved in the neural processes which underlie drug addiction. Several studies have reported that the administration of morphine induces changes in the levels and/or activity of endogenous opioid peptides (enkephalin, dynorphin) and their precursors in specific brain regions of the adult CNS. The aim of this work was to study the effects of chronic morphine exposure and its withdrawal on certain aminopeptidases capable of degrading opioid peptides in brain areas including the amygdala, hypothalamus, hippocampus, striatum and brain cortices. In animals treated with morphine, aminopeptidase N presented higher enzyme activity levels in the striatum, the hypothalamus and the amygdala compared to control animals, although statistically significant differences were observed only in the case of the striatum. In addition, the activity of soluble puromycin-sensitive aminopeptidase (PSA) was found to be higher in the frontal cortex of these rats. In contrast, rats experiencing withdrawal symptoms presented decreased levels of aminopeptidase activity in certain brain areas. Thus, the activity of aminopeptidase N in the hippocampus and soluble puromycin-sensitive aminopeptidase in the frontal cortex were found to be lower in rats experiencing naloxone precipitated withdrawal symptoms, compared to the corresponding controls. Finally, the activity of the three studied aminopeptidases in vitro was unaltered by incubation with morphine, suggesting that the observed effects are not due to a direct action of this opioid upon the aminopeptidases. The results of the present report indicate that aminopeptidases may play an important role in the processes of tolerance and withdrawal associated with morphine administration. PMID- 12573805 TI - Molecular cloning and functional characterization of a vasotocin receptor subtype expressed in the pituitary gland of the domestic chicken (Gallus domesticus): avian homolog of the mammalian V1b-vasopressin receptor. AB - The neurohypophysial hormone arginine vasotocin (AVT) stimulates adrenocorticotropin hormone (ACTH) secretion from the avian anterior pituitary gland resulting in increased adrenal secretion of corticosterone in response to stress. Here, we report molecular cloning and functional characterization of a gene encoding an AVT receptor subtype, designated the VT2 receptor, that may mediate the stimulatory effect of AVT on ACTH secretion in birds. The open reading frame predicts a 425 amino acid polypeptide that includes seven segments of 19 to 24 hydrophobic amino acids, typical of guanine nucleotide-protein coupled receptors. Phylogenetic analysis revealed that the VT2 receptor shares highest identity with the mammalian V1b-vasopressin receptor subtype. Expressed VT2 receptors in COS7 cells mediate AVT-induced phosphatidylinositol turnover and Ca(2+) mobilization. In the domestic chicken, expression of VT2 receptor gene transcripts is limited to the pituitary gland. Based on similarities in sequence, site of expression and coupled signal transduction pathways, we conclude that the VT2 receptor is the avian homolog of the mammalian V1b-vasopressin receptor, and therefore may play an important role in the avian stress response. PMID- 12573806 TI - Expression of angiotensin type 1 and 2 receptors in brain after transient middle cerebral artery occlusion in rats. AB - Angiotensin II (Ang II) type 2 receptors (AT2Rs) have been associated with apoptosis. We hypothesized that AT2Rs are increased in stroke and may contribute effects of stroke to the brain. To test this, we have examined the expression of Ang II type 1 receptor (AT1R), AT2R and Ang II levels in the brain 24 h after transient middle cerebral artery occlusion (MCAO). The densities of AT1R and AT2R were measured by quantitative autoradiography (n=6). The levels of Ang II were measured by radioimmunoassay (RIA) (n=6) and by immunohistochemistry (n=3). AT1R levels on autoradiography showed a significant decrease (0.87+/-0.06 to 1.39+/ 0.07 fmol/mg, p<0.01) in the ventral cortex of the stroke side compared to the cortices of non-stroke (NS) rats (n=4). There was no significant difference on ATIR in the contralateral verbal cortex of the stroke rats compared to NS control. In contrast, levels of AT2R in the ventral cortex of both the stroke and the contralateral sides were significantly increased (0.77+/-0.06, p<0.05 and 0.91+/-0.05, p<0.01 compared to 0.60+/-0.03 fmol/mg tissue, respectively). RIA showed that Ang II in the ventral cortex of both the stroke and the contralateral sides were significantly increased (241.63+/-47.72, p<0.01 and 165.51+/-42.59, p<0.05 compared to 76.80+/-4.10 pg/g tissue, respectively). Also, Ang II in the hypothalamus was significantly increased (179.50+/-17.49 to 118.50+/-6.65 pg/g tissue, p<0.05). Immunohistochemistry confirmed the increase of Ang II. These results demonstrate that brain Ang II and AT2Rs are increased whereas AT1Rs are decreased after transient MCAO in rats. We conclude that in stroke, Ang II and AT2R are activated and may contribute neural effects to brain ischemia. PMID- 12573807 TI - Trophic and steroidogenic effects of water deprivation on the adrenal gland of the adult female rat. AB - The effects of a 3-day water deprivation were studied in adult female rats in order to know what are the different zones of the adrenal gland and the hormonal factors involved in the growth and the activity of the adrenal gland. Water deprivation significantly increased plasma renin activity (PRA), plasma Angiotensin II (AII), vasopressin (AVP), epinephrine, aldosterone and corticosterone concentrations but did not modify the plasma adrenocorticotropin hormone (ACTH) level. Water deprivation significantly increased the absolute weight of the adrenal capsule containing the zona glomerulosa without modification of the density of cells per area unit suggesting that the growth of the adrenal capsule was due to a cell hyperplasia of the zona glomerulosa. Water deprivation significantly increased the density of AII type 1 (AT(1)) receptors in the adrenal capsule but did not modify the density of AII type 2 (AT(2)) receptors in the adrenal capsule and core containing the zona fasciculata, the zona reticularis and the medulla. The treatment of dehydrated female rats with captopril, which inhibits the angiotensin converting enzyme (ACE) in order to block the production of AII, significantly decreased the absolute weight of the adrenal capsule, plasma aldosterone and the density of AT(1) receptors in the adrenal capsule. The concentration of corticosterone in the plasma, the density of AT(2) receptors and the density of cells per unit area in the zona glomerulosa of the adrenal capsule were not affected by captopril-treatment. In conclusion, these results suggest that AII seems to be the main factor involved in the stimulation of the growth and the secretion of aldosterone by the adrenal capsule containing the zona glomerulosa during water deprivation. The low level of plasma ACTH is not involved in the growth of the adrenal gland but is probably responsible for the secretion of corticosterone by the zona fasciculata. PMID- 12573808 TI - Androgen physiology: unsolved problems at the millennium. AB - Androgen physiology differs from that of other steroid hormones in two major regards. First, testosterone, the predominant circulating testicular androgen, is both an active hormone and a prohormone for the formation of a more active androgen, the 5alpha-reduced steroid dihydrotestosterone. Genetic evidence indicates that testosterone and dihydrotestosterone work via a common intracellular receptor, and studies involving in vitro reporter gene assays and intact mice in which both steroid 5alpha-reductase isoenzymes have been disrupted by homologous recombination indicate that dihydrotestosterone acts during embryonic life to amplify hormonal signals that can be mediated by testosterone at higher concentrations. However, in post-embryonic life dihydrotestosterone plays unique roles that have not been elucidated. Studies of other 5alpha-reduced steroids, including the plant hormone brassinolide, the hog pheromones androstanol and androstenol, and 5alpha-dihydroprogesterone (in horses and elephants) indicate that this reaction serves different functions in different systems. Second, during embryonic life androgen causes the formation of the male urogenital tract and hence is responsible for development of the tissues that serve as the major sites of androgen action in postnatal life. It has been generally assumed that androgens virilize the male fetus by the same mechanisms as in the adult, namely by the conversion of circulating testosterone to dihydrotestosterone in target tissues. However, in marsupial mammals there is no sexual dimorphism in the levels of testosterone or dihydrotestosterone at the time the male phenotype forms, and in the pouch young of one marsupial, the tammar wallaby, the testes secrete another 5alpha-reduced steroid, 5alpha androstane-3alpha, 17beta-diol (5alpha-adiol), into plasma. The administration of 5alpha-adiol to female pouch young causes profound virilization of the urogenital sinus and external genitalia, but within target tissues 5alpha-adiol appears to work after oxidation to dihydrotestosterone. Thus, two separate mechanisms evolved for the formation of dihydrotestosterone in target tissues. 5alpha-adiol is the predominant androgen in neonatal testes in several placental mammals, but it is unclear whether it plays a similar role in other mammalian species. PMID- 12573810 TI - Molecular action of androgens. AB - The androgen receptor (AR), which mediates androgen action in the cell, belongs to the superfamily of nuclear receptors, a large group of transcription factors. Recent studies have described how the AR acts on specific target genes. The receptor's specificity of action depends on its regulation at different levels: expression in the cells, ligand binding and DNA-specific sequence recognition by structurally conserved domains and regulation by transcriptional factors in an integrated response. We propose, here, an overview of recent works on the molecular regulation of androgen-dependent genes by AR. PMID- 12573809 TI - Androgen biosynthesis from cholesterol to DHEA. AB - Androgens and estrogens are made from dehydroepiandrosterone (DHEA), which is made from cholesterol via four steps. First, cholesterol enters the mitochondria with the assistance of the steroidogenic acute regulatory protein (StAR). Mutations in the StAR gene cause congenital lipoid adrenal hyperplasia. Second, within the mitochondria, cholesterol is converted to pregnenolone by the cholesterol side chain cleavage enzyme, P450scc. Third, pregnenolone undergoes 17alpha-hydroxylation by microsomal P450c17. Finally, 17-OH pregnenolone is converted to DHEA by the 17,20 lyase activity of P450c17. The ratio of the 17,20 lyase to 17alpha-hydroxylase activity of P450c17 determines the ratio of C21 to C19 steroids produced. This ratio is regulated post-translationally by at least three factors: the abundance of the electron-donating protein P450 oxidoreductase, the presence of cytochrome b(5), and the serine phosphorylation of P450c17. Study of these and related factors may yield important information about the pathophysiology of adrenarche and the polycystic ovary syndrome (PCOS). PMID- 12573811 TI - Androgens and puberty. AB - Growth, adolescent sexual development and changes in body composition are described in male adolescents. The underlying changes in the hypothalamic pituitary gonadal and the GH/IGF-I axes that underline the physical changes are then described. Finally, how high energy output, at a time of low energy input, using, as an example, scholastic male wrestlers, is described. PMID- 12573812 TI - Androgens and male behavior. AB - Sexual differentiation into a male or a female includes sexual differentiation of the brain. The paradigm of mammalian sexual differentiation is that in the presence of androgens (normally produced by the fetal testis) a male brain differentiation occurs, while in the absence of androgens (normal in females) a female brain differentiation follows. In the human there is a sex-dimorphism in gender identity/role, sexual orientation, sexual functioning, and in non-sexual functions, such as spatial ability, and verbal fluency. Inasmuch these properties can be studied in other mammals the effects of androgens are solidly demonstrable. In the human the evidence for androgen effects is equally plausible, evident from observations in subjects with errors in the process of sexual differentiation and in morphological studies of brain structures presumably related to these properties. But clinical observations show compellingly that other, largely unidentified, factors may modulate, or even override the effects of androgens. PMID- 12573813 TI - Androgens and aging men. AB - This article underlines the hormonal and clinical events characterizing the so called andropause, as they may be described from an extensive review of the literature and some data obtained with the French colleagues of the author. Evidence-based hormonal intervention has to be positioned against anti-aging entrepreneurs who peddle hormones at random. The desequilibrium in aging men between the unchanged level of plasma cortisol and the profound decrease of androgens deserves cautious studies to eventually oppose this unbalanced hormonal situation effectively and safely. PMID- 12573814 TI - Androgens and male physiology the syndrome of 5alpha-reductase-2 deficiency. AB - Dihydrotestosterone (DHT), a potent androgen, is converted from testosterone by 5alpha-reductase isozymes. There are two 5alpha-reductase isozymes, type 1 and type 2 in humans and animals. These two isozymes have differential biochemical and molecular features. Mutations in type 2 isozyme cause male pseudohermaphroditism, and many mutations have been reported from various ethnic groups. The affected 46XY individuals have high normal to elevated plasma testosterone levels with decreased DHT levels and elevated testosterone/DHT ratios. They have ambiguous external genitalia at birth so that they are believed to be girls and are often raised as such. However, Wolffian differentiation occurs normally and they have epididymides, vas deferens and seminal vesicles. Virilization occurs at puberty frequently with a gender role change. The prostate in adulthood is small and rudimentary, and facial and body hair is absent or decreased. Balding has not been reported. Spermatogenesis is normal if the testes are descended. The clinical, biochemical and molecular genetic analyses of 5alpha reductase-2 deficiency highlight the significance of DHT in male sexual differentiation and male pathophysiology. PMID- 12573815 TI - Androgen receptor mutations and androgen insensitivity. AB - The androgen receptor (AR) is a high affinity receptor protein encoded on the human X-chromosome that mediates the actions of androgens during development and in the adult. Defects in this receptor protein result in a wide range of abnormalities of male sexual development. Studies in a number of different laboratories have identified mutations of the AR gene in subjects with androgen resistance syndromes. Defects that interrupt the AR open-reading frame have been traced to a number of distinct types of genetic alterations, have been identified in widely separated segments of the AR gene, and are invariably associated with the phenotype of complete androgen insensitivity. By contrast, mutations that cause single amino acid substitutions within the AR are localized to the DNA- or ligand-binding domains of the receptor protein and have been associated with the full range of androgen resistant phenotypes. The diversity of mutations that have been identified has prompted a consideration of the relationship between AR mutation and phenotype. Analyses of AR abundance and function suggest that the phenotypic abnormalities that result from mutation of the AR reflect the extent to which AR activity is impaired in target tissues. Such decreases in AR function may be the result of the diminished receptor function, decreases in receptor concentration, or a combination of these two effects. PMID- 12573816 TI - Des (1-3) IGF-I-stimulated growth of human stromal BPH cells is inhibited by a vitamin D3 analogue. AB - Prostate growth and differentiation is under the control of androgens not only during fetal life and childhood but also in adulthood, and it has been proposed that increased prostatic concentration of androgens, or increased androgen responsiveness, causes benign prostatic hyperplasia (BPH). However, different androgen ablation strategies such as treatment with GnRH agonists and finasteride resulted in a modest decrease of the hyperplastic prostate volume. In the last few years it became evident that both androgen-dependent and androgen-independent growth factors promote prostate enlargement by inducing cell proliferation or reducing apoptosis. Therefore, new therapeutic strategies, aimed at reducing intraprostatic growth factor signaling, are under investigation. In this study, we report further evidence that a non hypercalcemic-analogue of vitamin D(3), analogue (V) decreases growth factor-induced human BPH cell proliferation and survival. We found that Des (1-3) insulin-like growth factor [Des (1-3) IGF-I], an IGF-I analogue, which does not bind to IGF-binding proteins, is a potent mitogen for BPH stromal cells via a dual mechanism: stimulation of cell growth and inhibition of apoptosis. Similar results were previously reported for another growth factor for BPH cells, keratinocyte growth factor (KGF). Accordingly, we speculate that both KGF and IGF might be involved in the pathogenesis of BPH. We also found analogue (V) not only inhibits the mitogenic activity of growth factors on BPH cells, but even decreased the basal expression of bcl-2, and induced apoptosis. Therefore, vitamin D(3) analogues might be considered for the medical treatment of BPH. PMID- 12573817 TI - Androgen blockade in prostate cancer in 2002: major benefits on survival in localized disease. AB - The last 20 years have witnessed major advances in the field of prostate cancer, both in terms of diagnosis and treatment. Using screening with PSA, 99% of prostate cancers can now be diagnosed at a clinically localized or potentially curable stage. Over a 11-year period starting in 1988, the Quebec screening study performed among 45,000 men aged 45-80 years has shown that the prostate cancer death incidence has decreased by 64% in men who had screening. The impact of screening, however, requires early application of the most efficacious treatments. In this context, the most important recent therapeutic advance in the field of prostate cancer is androgen blockade, namely medical castration with LHRH agonists, the availability of pure antiandrogens and combined androgen blockade (CAB) using medical or surgical castration in association with a pure antiandrogen. In the six studies performed in localized or locally advanced disease, the improved cancer-specific survival ranges between 37 and 81% at 5 years of follow-up for patients who received androgen blockade. On the other hand, data already available show that long term and continuous (not intermittent) androgen blockade is highly efficient and can achieve long term control or possible cure of localized prostate cancer. PMID- 12573818 TI - Androgens and alopecia. AB - Androgens have profound effects on scalp and body hair in humans. Scalp hair grows constitutively in the absence of androgens, while body hair growth is dependent on the action of androgens. Androgenetic alopecia, referred to as male pattern hair loss (MPHL) in men and female pattern hair loss (FPHL) in women, is due to the progressive miniaturization of scalp hair. Observations in both eunuchs, who have low levels of testicular androgens, and males with genetic 5alpha-reductase (5alphaR) deficiency, who have low levels of dihydrotestosterone (DHT), implicate DHT as a key androgen in the pathogenesis of MPHL in men. The development of finasteride, a type 2-selective 5alphaR inhibitor, further advanced our understanding of the role of DHT in the pathophysiology of scalp alopecia. Controlled clinical trials with finasteride demonstrated improvements in scalp hair growth in treated men associated with reductions in scalp DHT content, and a trend towards reversal of scalp hair miniaturization was evident by histopathologic evaluation of scalp biopsies. In contrast to its beneficial effects in men, finasteride did not improve hair growth in postmenopausal women with FPHL. Histopathological evaluation of scalp biopsies confirmed that finasteride treatment produced no benefit on scalp hair in these women. These findings suggest that MPHL and FPHL are distinct clinical entities, with disparate pathophysiologies. Studies that elucidate the molecular mechanisms by which androgens regulate hair growth would provide greater understanding of these differences. PMID- 12573819 TI - Antiandrogens: selective androgen receptor modulators. AB - Antiandrogens can efficiently block androgen receptor (AR) mediated gene expression, and are therefore useful tools in the treatment of androgen dependent prostate cancer. Antiandrogens are either complete or partial inhibitors of AR activity, depending on the nature of the compound. As compared to androgens, antiandrogens induce a different AR conformation, thereby influencing the recruitment of co-regulators (coactivators and corepressors). This ligand selective modulation of AR activity is affected by an AR mutation (Thr877Ala substitution) found in prostate cancer. In contrast to the wild-type AR, the mutant AR conformation induced by cyproterone acetate (CPA) and hydroxyflutamide (OHF) is comparable to that induced by androgens. As a consequence, this might affect recruitment of co-regulators, thereby allowing CPA and OHF to act as strong agonists on the mutant AR. PMID- 12573820 TI - Potential action of IGF-1 and EGF on androgen receptor nuclear transfer and transactivation in normal and cancer human prostate cell lines. AB - This work was designed to determine whether IGF-1 and EGF modulate nuclear transfer and transactivation of the androgen receptor (AR) in human prostate cell lines (PNT1A and DU-145). We first characterized the IGF-1 and EGF receptors by ligand-binding assays with [125I] IGF-1 and [125I] EGF in a normal human prostate epithelial cell line, PNT1A. We then evaluated the effects of these growth factors on AR nuclear transfer and transcriptional activation in this cell line and in DU-145, a human prostate tumor cell line. The cell lines were cotransfected with an AR expression vector and an androgen-responsive luciferase gene driven by the mouse mammary tumor virus (MMTV-luciferase) promoter. Neither IGF-1 nor EGF could activate reporter gene in the absence of androgens. Conversely, both enhanced the magnitude of the AR response in the presence of low levels of androgen (10(-11)-10(-9) M) and this response, increased by twofold, was inhibited by hydroxyflutamide. No effect of IGF-1 and EGF was observed on the intracellular localization of the fusion protein EGFP-AR in either cell line. The fluorescence stayed cytoplasmic even after 24 h of IGF-1 or EGF treatment. Taken together, these data indicate that growth factors are unable to initiate the nuclear translocation of AR in the absence of androgens or to induce ligand independent transcriptional activity. We observed only cross-talk in the presence of androgens and IGF-1 or EGF, leading to an over-activated AR. In conclusion, the cross-talk between AR and growth factor signaling pathways may sensitize AR to suboptimal stimulation by low levels of androgens. PMID- 12573821 TI - Identification of tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) and its receptors in adult rat ventral prostate. AB - Tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor-alpha (TNF-alpha) family of cytokines that is known to induce apoptosis upon binding to its death domain-containing receptors, DR4/TRAIL-R1 and DR5/TRAIL-R2. Two additional TRAIL receptors, DcR1/TRAIL-R3 and DcR2/TRAIL-R4, lack functional death domains and act as decoy receptors for TRAIL. In this study, the presence of TRAIL and its receptors was investigated in adult rat hormonosensitive ventral prostate. TRAIL and its receptors were identified in the rat ventral prostate in terms of protein and mRNA. TRAIL and its receptors were immunolocalized in prostatic epithelial cells. PMID- 12573822 TI - Evaluation of androgenic bioactivity in human serum by recombinant cell line: preliminary results. AB - Besides the measurement of circulating conjugated metabolites of dihydrotestosterone (DHT), which reflects androgenic activity, only one assay to measure androgenic bioactivity in human serum has been proposed thus far. This recombinant bioassay is based on the androgen-dependent interaction between the LBD and NT domains of AR fused to the Gal 4 DNA-binding domain, but its construction is highly complex. We have developed a mammalian cell (CHO 515) bioassay that measures total androgen bioactivity in human serum. The AR deficient Chinese hamster ovary (CHO) cells were stably transfected with pSG5 puro-hAR and pMMTV-neo-Luc. After selection with puromycin and neomycin, five highly inducible clones were isolated and one was selected. The expression of human androgen receptor (hAR) was confirmed by Western blot and steroid-binding assays on the whole cells. The transcriptional activity of the clone was measured after 24 h of incubation with increasing concentrations of various androgenic and non-androgenic steroid compounds in a 96-well plate. The EC50s for each tested androgenic steroid compound were 4 x 10(-11) M, 1.5 x 10(-11) M, 1 x 10(-9) M, 2 x 10(-10) M, 3 x 10(-10) M for testosterone, DHT, dehydroepiandrosterone (DHEA), delta5-androstenediol, and delta4-androstenedione, respectively. In the physiological concentrations of the non-androgenic steroids, estradiol, cortisol, aldosterone, and progesterone, no interference was noted with the AR transactivation level. Evaluation of androgenic bioactivity in human serum was performed by incubation of CHO 515 cells with 100 microl of patient serum, diluted at 1/100 = 1% in DMEM-F12 without phenol red. The sensitivity of the assay was < 0.3 ng/ml. The mean androgenic bioactivity expressed in testosterone equivalents was 0.6 +/- 0.2 ng/ml in normal prepubertal boys, and 12.4 +/- 2 and 1.7 +/- 0.1 ng/ml in normal pubertal boys and girls, respectively. In conclusion, this new recombinant cell bioassay is today the only assay that takes into account testosterone, DHT, DHEA, delta5-androstenediol, and delta4 androstenedione. It should be of particular use in male children with cryptorchidism, delayed puberty or hypogonadotrophic hypogonadism, i.e., in pediatric patients with low androgen levels. PMID- 12573823 TI - Effects of androgen-binding protein (ABP) on spermatid Tnp1 gene expression in vitro. AB - In vitro studies were designed to determine whether Sertoli cell-delivered ABP could act on spermatogenetic events, whether such an action could occur via a paracrine or a juxtacrine pathway and whether sex steroids could be involved in this action. ABP delivery to germ cells was achieved using an in vitro model based on recombinant rat ABP-producing mouse Sertoli cells cocultivated with rat spermatids. Using semi-quantitative RT-PCR, the expression of the Tnp 1 gene encoding the Transition Protein 1, involved in the histone to protamine replacement during spermatid nuclear transformation, was analyzed. Our results provide clear evidence that Sertoli cell-derived ABP acts on spermatids by modifying the TP1 mRNA level. This outcome, strictly requiring juxtacrine conditions, is obtained in the absence of sex steroid hormones. To our knowledge this is the first evidence of an effect of ABP itself on male germ cells. PMID- 12573824 TI - Evidence that chlormadinone acetate exhibits antiandrogenic activity in androgen dependent cell line. AB - Chlormadinone acetate (CMA), like other 17-hydroxyprogesterone derivatives, is thought to be a potential antiandrogen on the basis of its effect on spontaneous benign prostatic hyperplasia (BPH) in dogs. This work was undertaken to find out whether CMA presents antiandrogen activity in human androgen-dependent cell line. For this purpose, we used PALM cells, the PC-3 cell line stably transfected with human androgen receptor and a luciferase gene under transcriptional control of MMTV. Potential antiandrogenic activity was compared with that of cyproterone acetate (CPA), a standard steroidal antiandrogen. Both compounds were tested in competitive binding assays at 37 degrees C in the presence of 1 nM of [3H] R1881, a synthetic and non-metabolizable androgen. Their impact on AR transcriptional activity was evaluated by the measure of luciferase activity in the presence of R1881 with increasing concentrations of CMA or CPA (10(-8)-10(-6) M). In whole cell binding assays, competitive studies revealed that the Ki for CMA was 3.3 +/- 1.5 x 10(-8) M (versus 7.2 +/- 1.3 x 10(-8) M for CPA). Inhibition of AR transcriptional activity was 40 +/- 5% for CMA (3 x 10(-7) M) versus 59 +/- 6% for CPA at the same concentration. Moreover, CMA caused a slower import of green fluorescent protein (GFP)-AR to the nuclei of COS-7 cells than R1881. These data show that CMA exerted a competitive binding for AR and significantly decreased the AR transcriptional activity. In conclusion, this synthetic progestin presents simultaneous antiandrogenic activity that could be helpful as a new therapeutic option in women with luteal defect along with clinical signs of hyperandrogenism. PMID- 12573825 TI - An investigation of iron sulphide dust minimum ignition temperatures. AB - An investigation of the ignition behaviour of iron sulphide dusts has been undertaken. Commercial samples of FeS and FeS(2) and mine samples of pyrrhotite and pyrite were tested for minimum ignition temperature (MIT) using a device known as the BAM oven. The mine samples were found to undergo a decrease in MIT as the mass mean particle diameter became smaller. Using available theoretical treatments, this experimental observation was interpreted as providing further evidence of the importance of heterogeneous combustion in the ignition of iron sulphide dusts. A dense cloud state was proven for the experimental apparatus used, and an alternate criterion for the boundary between a dilute and a dense dust cloud was proposed in terms of the number of dust particles present in the cloud. PMID- 12573826 TI - Master Logic Diagram: method for hazard and initiating event identification in process plants. AB - Master Logic Diagram (MLD), a method for identifying events initiating accidents in chemical installations, is presented. MLD is a logic diagram that resembles a fault tree but without the formal mathematical properties of the latter. MLD starts with a Top Event "Loss of Containment" and decomposes it into simpler contributing events. A generic MLD has been developed which may be applied to all chemical installations storing toxic and/or flammable substances. The method is exemplified through its application to an ammonia storage facility. PMID- 12573827 TI - A simple model for the release rate of hazardous gas from a hole on high-pressure pipelines. AB - It is very important to estimate the mass flow rate of possible accidental releases from the gas pipeline, in order to perform the hazard analysis or the risk based management in the gas facilities. This paper presents a simplified model to estimate the release rate from a hole on the high-pressure gas pipeline. It consists of a correction factor accounting for the pressure drop through pipeline due to the wall friction loss, and the release rate without friction loss. The model, whatever kind of gas may be considered, has some positive deviation from the theoretical complex equations, and it ranges from about 0 up to 20%. The deviation will reduce to zero, as the release point approaches to the reservoir. It increases with the specific heat ratio of gas and the dimensionless hole-size which is the effective area of the hole divided by the cross-sectional area of the pipe. The model is compared with damage areas of real accidents with success. It overestimates the release rate slightly and may be a useful tool to estimate the release rate quickly in performing the hazard analysis or the risk based management in the gas facilities. PMID- 12573828 TI - Adsorption of nickel(II) from aqueous solution onto activated carbon prepared from almond husk. AB - Activated carbon was prepared from almond husk by activating without (MAC-I) and with (MAC-II) H(2)SO(4) at different temperatures. The ability of the activated carbon to remove nickel(II) ions from aqueous solutions by adsorption has been investigated under several conditions such as pH, carbonisation temperature of husk, initial concentration of metal ions, contact time, and adsorbent concentration. Optimal conditions were pH 5.0, the carbonisation temperature of 700 degrees C, 50 min of contact time and adsorbent concentration of 5 g/l. The results indicate that the effective uptake of Ni(II) ions was obtained by activating the carbon, prepared from almond husk at 700 degrees C, through the addition of H(2)SO(4). The removal of Ni(II) were found to be 97.8% at initial concentration of 25mg/l and the adsorbent concentration of 5 g/l. When the adsorbent concentration was increased up to 40 g/l, the adsorption density decreased from 4.89 to 0.616 mg/g for MAC-II. In the isotherm studies, the experimental adsorption data fitted reasonably well the Langmuir isotherm for both MAC-I and MAC-II. PMID- 12573829 TI - On-site treatment and landfilling of MSWI air pollution control residues. AB - Air pollution control (APC) residues from municipal solid waste incineration (MSWI) are difficult to landfill due to substantial leaching of trace metals. An on-site pretreatment prior to landfilling of APC-residues was investigated in terms of bench-scale experiments with a semidry APC-residue and a fly ash. The treatment involved mixing of the residues with a ferrous sulphate solution and subsequent oxidation of the suspension. Afterwards, the suspension was spread on a dedicated landfill section and allowed to drain by gravity through the drainage system of the landfill. The wastewater from the process, collected through the drainage system, contained large concentrations of salts (Cl: 14-30 g/l, Na: 4-9 g/l, K: 5-11 g/l, Ca: 2-12 g/l) but low concentrations of trace metals (e.g. Pb: 14-100 microg/l, Cd: <2-7 microg/l). The treated residues left in the landfills were later subject to leaching by simulated rainfall. The leachate contained low concentrations of trace metals (Pb: <120 microg/l, Cd: <2 microg/l, Cr: <485 microg/l). The leachate concentrations from the treated APC-residues were substantially reduced compared to concentrations in leachate from untreated APC residues. Particularly in the early stages of the leaching, concentrations of trace metals were reduced by up to four orders of magnitude. PMID- 12573830 TI - Thorium removal by different adsorbents. AB - The removal of radiotoxic Th(4+) from aqueous solutions has been explored using two different groups of adsorptive materials (e.g. two activated carbons and four zeolites-two natural and two synthetic). The activated carbons were prepared from solvent extracted olive pulp (SEOP) and olive stone (OS) by a two-step physical activation method with steam. They were characterized by N(2) at 77K adsorption, Hg porosimetry and by determination of their iodine number. All carbons prepared are of the H-type (e.g. contain mainly basic surface oxides) confirmed by the results of the Boehm's method. The natural zeolites, clinoptilolite (NaCLI) and mordenite (NaMOR), were pretreated with Na(+) before the adsorption experiments, while the synthetic ones, NaX and NaA, were provided in their commercial sodium form. The natural zeolites, NaCLI and NaMOR, utilized 11.5 and 38.6% of the theoretical ion-exchange capacity, based on Al content, respectively, while NaX and NaA utilized 41.5 and 45.9%, respectively. The activated carbons showed better removal capability than NaCLI. NaMOR, showed comparable results to the carbon originated from OS, but lower removal capability than the carbon originated from SEOP. The synthetic zeolites showed the highest removal ability for thorium ions due to their increased ion-exchange capacity because of their cleaner and larger framework channels and their higher number of ion-exchange sites. The carbons adsorption capacity mainly depends on the content and nature of functional surface groups. The adsorption data were fitted to Langmuir and Freundlich models. The former achieved best fits and was further applied to obtain the respective Langmuir constant and maximum adsorption capacity for each system. PMID- 12573831 TI - Spectral fluorescence signatures and partial least squares regression: model to predict dissolved organic carbon in water. AB - Spectro-fluorescence signature (SFS) of water samples contains information that may be used to quantify dissolved organic carbon (DOC) if combined with multivariate analyses. A model was built through SFS and partial least squared (PLS) regression. The SFSs of 219 samples of natural water along the Raritan River and Millstone River watersheds located in central New Jersey, and their corresponding DOC concentrations were used to build the model. Calibration, full cross-validation, and prediction performances of various models were statistically compared before optimal model selection. The final selected model, tested on the Passaic River watershed in northern New Jersey, provided a bias of 0.028 mg/l and a root mean squared error of prediction (RMSEP) of 0.35 mg/l. Linked to PLS, SFS can be a quality and cost effective method to perform on-line rapid DOC measurements. PMID- 12573832 TI - Elution and decomposition of cyanide in soil contaminated with various cyanocompounds. AB - Standard soil samples contaminated with various standard cyanocompounds were prepared. Column elution experiments and analyses were conducted. Compounds with an easy capacity for dissociation to ions, such as KCN and potassium hexacyanoferrate(III), were found to be eluted by forming free cyanide even in fresh water. Hexacyanoferrate(II) salts, such as potassium hexacyanoferrate(II) and iron(III) hexacyanoferrate(II), were found not to be dissociated in water, but were dissociated and diffused under alkaline conditions (pH >13). Hexacyanoferrate(II) ion was found to be more easily dissociated in water with a higher pH. Column tests as above were also conducted for soil samples taken from a former paint ink factory using iron(III) hexacyanoferrate(II), cyanogen chloride, potassium cyanate, copper cyanide, as well as potassium cyanide, as raw materials. It was demonstrate that iron(III) hexacyanoferrate(II) was dissociated and eluted under alkaline conditions. The elution rate was reduced when the contaminated soil was sandwiched with standard soil layers.Further, it was found that the Fe(CN)(6)(4-) ion eluted with NaOH from hexacyanoferrate acid in soil, were easily decomposed into cyanic acid or other byproducts by UV with the addition of ozone and H(2)O(2). PMID- 12573833 TI - Procion Green H-4G immobilized poly(hydroxyethylmethacrylate/chitosan) composite membranes for heavy metal removal. AB - The effective removal of toxic heavy metals from environmental samples still remains a major topic of present research. Metal-chelating membranes are very promising materials as adsorbents when compared with conventional beads because they are not compressible, and they eliminate internal diffusion limitations. The purpose of this study was to evaluate the performance of a novel adsorbent, Procion Green H-4G immobilized poly(hydroxyethylmethacrylate (HEMA)/chitosan) composite membranes, for the removal of three toxic heavy metal ions, namely, Cd(II), Pb(II) and Hg(II) from aquatic systems. The Procion Green H-4G immobilized poly(hydroxyethylmethacrylate/chitosan) composite membranes were characterized by elemental analysis, scanning electron microscopy and Fourier transform infrared (FTIR) spectroscopy. The immobilized amount of the Procion Green H-4G was calculated as 0.018+/-0.003 micromol/cm(2) from the nitrogen and sulphur stoichiometry. The adsorption capacity of Procion Green H-4G immobilized poly(hydroxyethylmethacrylate/chitosan) composite membranes for selected heavy metal ions from aqueous media containing different amounts of these ions (30 400mg/l) and at different pH values (2.0-6.0) was investigated. The amount of Cd(II), Pb(II) and Hg(II) adsorbed onto the membranes measured at equilibrium, increased with time during the first 45 min and then remained unchanged toward the equilibrium adsorption. The maximum amounts of heavy metal ions adsorbed were 43.60+/-1.74, 68.81+/-2.75 and 48.22+/-1.92 mg/g for Cd(II), Pb(II) and Hg(II), respectively. The heavy metal ion adsorption on the pHEMA/chitosan membranes (carrying no dye) were relatively low, 6.31+/-0.13 mg/g for Cd(II), 18.73+/-0.37 mg/g for Pb(II) and 18.82+/-0.38 mg/g for Hg(II). Competitive adsorption of the metal ions was also studied. When the metal ions competed with each other, the adsorbed amounts were 12.74+/-0.38 mg Cd(II)/g, 28.80+/-0.86 mg Pb(II)/g and 18.41+/-0.54 mg Hg(II)/g. Procion Green H-4G immobilized poly(hydroxyethylmethacrylate/chitosan) membranes can be regenerated by washing with a solution of nitric acid (0.01 M). The percent desorption achieved was as high as 95%. These novel membranes are suitable for repeated use for more than five adsorption/desorption cycles without any considerable loss in adsorption capacity. Adsorption equilibria were well described by Langmuir equation. It can be concluded that Procion Green H-4G immobilized poly(hydroxyethylmethacrylate/chitosan) membranes may effectively be used for the removal of Cd(II), Pb(II) and Hg(II) ions from aqueous solutions. PMID- 12573834 TI - Vacuum extraction based response equipment for recovery of fresh fuel spills from soil. AB - Accidental overturns of fuel tankers can have, depending on soil types, severe consequences. This applies, particularly in areas of shallow soils where the groundwater is located 2-4m below the ground surface. By rapid, vacuum extraction based recovery emergency services, which would normally be the first to arrive on the scene, could minimize consequences of fresh fuel spills and even prevent groundwater contamination, the primary purpose of emergency response. Powerful vacuum extraction-based response (PER), equipment has been developed to recover freshly spilt volatile fuels from the soil, primary by emergency services, but also by other trained responders. The main components of mobile PER-equipment are perforated extraction pipes, a recovery vacuum tank, a vacuum pump and an incinerator. The PER-equipment has been tested in summer and sub-zero winter conditions, and in both cases 50-80% of fresh gasoline spilled into sandy soil was recovered during the first 2h of operation. Gasoline was recovered in both liquid and vapor form, and hydrocarbon vapors were destroyed by controlled incineration at a safe distance from the spill. Recovery of less volatile diesel oil is not so effective from the sandy soil, but about 30% of it could be pumped from a fresh pool directly after a seepage time of 15 min. PMID- 12573835 TI - Emission control of SO2 and NOx by irradiation methods. AB - Microwave discharges at 2.45 GHz frequency and accelerated electron beams operated at atmospheric pressure in synthetic gas mixtures containing N(2), O(2), CO(2), SO(2), and NO(x) are investigated experimentally for various gas mixture constituents and operating conditions, with respect to their ability to purify exhaust gases. An original experimental unit easily adaptable for both separate and simultaneous irradiation with microwaves and electron beams was set up. The simultaneous treatment with accelerated electron beams and microwaves was found to increase the removal efficiency of NO(x) and SO(2) and also helped to reduce the total required dose rate with approximately 30%. Concomitant removal of NO(x) ( approximately 80%) and SO(2) (>95%) by precipitation with ammonia was achieved. PMID- 12573836 TI - Combined physical, chemical and biological treatments of wastewater containing organics from a semiconductor plant. AB - Wastewater containing organics from a semiconductor plant was experimentally investigated in this study. The wastewater is characterized by strong color, high chemical oxygen demand (COD), a large amount of refractory volatile organic compounds and low biodegradability. Because of these characteristics, treatment of this wastewater by traditional activated sludge method is essentially impossible. In the present work, combined physical, chemical and biological methods were synergistically utilized to tackle the wastewater. The combined treatment consisted of air stripping, modified Fenton oxidation and sequencing batch reactor (SBR) method. Air stripping was employed to remove the majority of volatile organic components (notably isopropyl alcohol) from the wastewater, while the Fenton treatment decomposed the remaining refractory organics leading to simultaneous reductions of wastewater COD and color. After proper dilution with other low-strength, organics-containing wastewater stream, the wastewater effluent was finally treated by the SBR method. Experimental tests were conducted to determine the effectiveness and the optimum operating conditions of each treatment process. Test results clearly demonstrated the advantages of the combined treatments. The treatment train was found capable of lowering the wastewater COD concentration from as high as 80,000 mg/l to below 100mg/l and completely eliminating the wastewater color. The overall water quality of the final effluent exceeded the direct discharge standard and the effluent can even be considered for reuse. PMID- 12573837 TI - Detoxification of a highly toxic lead-loaded industrial solid waste by stabilization using apatites. AB - Apatites are known for their properties to immobilize lead contained in aqueous solutions or contaminated soils. In this study, apatites were examined as stabilization additives for lead-loaded industrial solid toxic wastes. The specific waste was the residue, obtained after thermal treatment of sludges (incineration), which was derived from tetraethyl lead fuel storage tanks. It was found to contain around 30 wt.% lead and 33 wt.% iron. Standard leaching tests (according to DIN 38414 S-4) were applied for the determination of leachability of metals from the ash and, thus, of chemical toxicity; the proposed leaching tests examined both initial and stabilized products in order to evaluate the effectiveness of the applied additives. The results obtained demonstrate the fact that lead concentrations in leachates, after the application of the proposed leaching tests using apatites as additives and with a ratio of 50% solid waste-50 wt.% apatite, could be reduced to the range of 1mg/l. PMID- 12573838 TI - Characterisation of phosphorous forms in wastewater treatment plants. AB - The removal of different forms of phosphorous (namely total phosphorous, soluble phosphorous, particulate phosphorous and total phosphate) has been studied in two municipal wastewater treatment plants (WWTP) with different characteristics, but without any specific implemented strategy for phosphorous removal. The results obtained for the different forms of phosphorus can be summarised as follows: (1) complete removal of particulate phosphorous is achieved in either primary or secondary clarifiers; (2) total phosphorous concentration in the effluent is mostly soluble phosphorous and this is mainly phosphate; (3) a small amount of soluble phosphorous is removed by biomass growth and/or biosorption; (4) both WWTPs presented a high-buffered behaviour in response to high inlet loading of phosphorous, showing a constant pattern at the outlet of the WWTP; (5) removal of total phosphorous was approximately 60-70% for both WWTPs; and (6) recirculation streams such as supernatant from centrifuge sludge dehydration operation can have a significant contribution to the inlet amount of phosphorous. The results presented in this paper provide a basis to develop prospects for phosphorous removal, which may be adapted to the particular configurations of the WWTP studied. PMID- 12573839 TI - Utilization of ICP/OES for the determination of trace metal binding to different humic fractions. AB - In this study, the use of inductively coupled plasma/optical emission spectrometry (ICP/OES) to determine multi-metal binding to three biomasses, Sphagnum peat moss, humin and humic acids is reported. All the investigations were performed under part per billion (ppb) concentrations. Batch pH profile experiments were performed using multi-metal solutions of Cd(II), Cu(II), Pb(II), Ni(II), Cr(III) and Cr(VI). The results showed that at pH 2 and 3, the metal affinity of the three biomasses exposed to the multi-metal solution that included Cr(III) presented the following order: Cu(II), Pb(II)>Ni(II)>Cr(III)>Cd(II). On the other hand, when Cr(VI) was in the heavy metal mixture, Sphagnum peat moss and humin showed the following affinity: Cu(II), Pb(II)>Ni(II)>Cr(VI)>Cd(II); however, the affinity of the humic acids was: Cu(II)>Pb(II), Cr(VI)>Ni(II)>Cd(II). The results demonstrated that pH values of 4 and 5 were the most favorable for the heavy metal binding process. At pH 5, all the metals, except for Cr(VI), were bound between 90 and 100% to the three biomasses. However, the binding capacity of humic acids decreased at pH 6 in the presence of Cr(VI). The results showed that the ICP/OES permits the determination of heavy metal binding to organic matter at ppb concentration. These results will be very useful in understanding the role of humic substances in the fate and transport of heavy metals, and thus could provide information to develop new methodologies for the removal of low concentrations of toxic heavy metals from contaminated waters. PMID- 12573840 TI - Low-cost adsorbents for heavy metals uptake from contaminated water: a review. AB - In this article, the technical feasibility of various low-cost adsorbents for heavy metal removal from contaminated water has been reviewed. Instead of using commercial activated carbon, researchers have worked on inexpensive materials, such as chitosan, zeolites, and other adsorbents, which have high adsorption capacity and are locally available. The results of their removal performance are compared to that of activated carbon and are presented in this study. It is evident from our literature survey of about 100 papers that low-cost adsorbents have demonstrated outstanding removal capabilities for certain metal ions as compared to activated carbon. Adsorbents that stand out for high adsorption capacities are chitosan (815, 273, 250 mg/g of Hg(2+), Cr(6+), and Cd(2+), respectively), zeolites (175 and 137 mg/g of Pb(2+) and Cd(2+), respectively), waste slurry (1030, 560, 540 mg/g of Pb(2+), Hg(2+), and Cr(6+), respectively), and lignin (1865 mg/g of Pb(2+)). These adsorbents are suitable for inorganic effluent treatment containing the metal ions mentioned previously. It is important to note that the adsorption capacities of the adsorbents presented in this paper vary, depending on the characteristics of the individual adsorbent, the extent of chemical modifications, and the concentration of adsorbate. PMID- 12573841 TI - Mass transfer coefficient and concentration boundary layer thickness for a dissolving NAPL pool in porous media. AB - Analytical expressions for the time invariant, average mass transfer coefficient and the concentration boundary layer thickness applicable to dissolving single component nonaqueous phase liquid (NAPL) pools in two-dimensional, saturated, homogeneous and isotropic porous formations are derived. Good agreement between predicted and experimentally determined time invariant average mass transfer coefficients is observed. PMID- 12573842 TI - Treatment of industrial oily wastewaters by wet oxidation. AB - In the present work, the homogeneous wet oxidation (WO) of an oily wastewater (COD approximately 11,000 mg l(-1)), composed mainly of alcohols and phenolic compounds, was studied in a high-pressure agitated autoclave reactor in the temperature range of 180-260 degrees C and oxygen pressure 1 MPa. Temperature was found to have a significant impact on the oxidation of the contaminants in the wastewater. Among the compounds contained in the wastewater, ethylene glycol showed great resistance to wet oxidation. Temperatures above 240 degrees C were required for its effective degradation. Organic acids, mainly acetic acid, were the intermediate products of the wet oxidation process and their conversion to carbon dioxide was very slow. A generalised model based on a parallel reaction scheme was used to interpret the experimental data obtained. The activation energies obtained were in the range of 90-130 kJ mol(-1). PMID- 12573843 TI - Atomization of liquids in a Pease-Anthony Venturi scrubber. Part I. Jet dynamics. AB - Jet dynamics, in particular jet penetration, is an important design parameter affecting the collection efficiency of Venturi scrubbers. A mathematical description of the trajectory, break-up and penetration of liquid jets initially transversal to a subsonic gas stream is presented. Experimental data obtained from a laboratory scale Venturi scrubber, operated with liquid injected into the throat through a single orifice, jet velocities between 6.07 and 15.9 m/s, and throat gas velocities between 58.3 and 74.9 m/s, is presented and used to validate the model. PMID- 12573844 TI - Kinetic study of the gas-phase hydrogenation of aromatic and aliphatic organochlorinated compounds using a Pd/Al2O3 catalyst. AB - The hydrodechlorination of dichloromethane (DCM), tetrachloroethylene (TTCE), chlorobenzene (CBZ) and 1,2-dichlorobenzene (DCBZ), all of them alone and in mixtures, over a commercial Pd/Al(2)O(3) catalyst in a continuous packed-bed reactor was studied in the present work. Results indicate that the reaction kinetics for the single compounds are pseudo-first order kinetics. The reactivity of the compounds studied is very different. So, whereas aromatic compounds and tetrachloroethylene can be fully converted at the operation conditions reported in this work, dichloromethane conversions are lower than 30% in all the cases. The hydrodechlorination of mixtures of organochlorinated compounds shows important inhibition effects, these effects increase as the number of chlorine atoms in the molecule increase. Reaction kinetics for the hydrogenation of mixtures can be represented by a Langmuir-Hinshelwood model. PMID- 12573845 TI - Alkaline thermal sludge hydrolysis. AB - The waste activated sludge (WAS) treatment of wastewater produces excess sludge which needs further treatment prior to disposal or incineration. A reduction in the amount of excess sludge produced, and the increased dewaterability of the sludge are, therefore, subject of renewed attention and research. A lot of research covers the nature of the sludge solids and associated water. An improved dewaterability requires the disruption of the sludge cell structure. Previous investigations are reviewed in the paper. Thermal hydrolysis is recognized as having the best potential to meet the objectives and acid thermal hydrolysis is most frequently used, despite its serious drawbacks (corrosion, required post neutralization, solubilization of heavy metals and phosphates, etc.). Alkaline thermal hydrolysis has been studied to a lesser extent, and is the subject of the detailed laboratory-scale research reported in this paper. After assessing the effect of monovalent/divalent cations (respectively, K(+)/Na(+) and Ca(2+)/Mg(2+)) on the sludge dewaterability, only the use of Ca(2+) appears to offer the best solution. The lesser effects of K(+), Na(+) and Mg(2+) confirm previous experimental findings. As a result of the experimental investigations, it can be concluded that alkaline thermal hydrolysis using Ca(OH)(2) is efficient in reducing the residual sludge amounts and in improving the dewaterability. The objectives are fully met at a temperature of 100 degrees C; at a pH approximately 10 and for a 60-min reaction time, where all pathogens are moreover killed. Under these optimum conditions, the rate of mechanical dewatering increases (the capillary suction time (CST) value is decreased from approximately 34s for the initial untreated sample to approximately 22s for the hydrolyzed sludge sample) and the amount of DS to be dewatered is reduced to approximately 60% of the initial untreated amount. The DS-content of the dewatered cake will be increased from 28 (untreated) to 46%.Finally, the mass and energy balances of a wastewater treatment plant with/without advanced sludge treatment (AST) are compared. The data clearly illustrate the benefits of using an alkaline AST-step in the system. PMID- 12573846 TI - Oil mobility in a saturated water-wetted bed of glass beads. AB - The mobilization of an oil bank in a packed bed of glass beads saturated with an aqueous phase has been studied both theoretically and experimentally. The size of the glass beads was varied in the range between 0.5 and 5mm. Two oils (hexadecane and hexane) with viscosities different for an order of magnitude and densities smaller than that of water have been used. A few more runs have been carried out using perchloroethylene (PCE), with density greater than that of water. The interfacial tension in the aqueous phase was varied in a quite large range (0.38 39.1 dyn/cm) by adding surfactants to the water. The glass assembly made it possible to follow the evolution of the dyed oily phase by the use of a digital camera. A very simple stochastic model for describing the porous structure of the packed bed made it possible to set a criterion for determining the probability of mobilization of ganglia which are produced by the fragmentation of the oil bank. The same model permits also to estimate the probability function of the velocity of a ganglion of an assigned size. PMID- 12573850 TI - Inducible systems see the light. AB - Advances in our capacity to design and use novel strategies for achieving inducible gene expression will improve our ability to define gene function. An extremely efficient system designed by nature -- that of the regulatable phytochrome system in plants -- has provided the basis for developing a novel inducible gene expression system. PMID- 12573851 TI - The new role of SAGE in gene discovery. AB - The sequencing of the human genome has led to in silico predictions of far fewer genes than anticipated. Recent studies using serial analysis of gene expression have cast doubt on this finding. One study predicts the presence of many unidentified low abundance transcripts, whereas two others have mapped unique tags to previously unpredicted exons. Genome and transcriptome complexity is thus greater than predicted and many of the missing genes are probably expressed in low copy numbers or only in early embryonic tissues. PMID- 12573853 TI - 21st century wheat breeding: plot selection or plate detection? AB - The publicly reported limited application of marker-assisted selection (MAS) in wheat breeding programmes to date is reviewed and contrasted with the current situation, in which it has increasingly become technically feasible to tag almost any gene with a microsatellite assay. Although this capability is starting to have an impact on the conduct of large breeding programmes, a much more profound change in breeding strategy will become possible when single nucleotide polymorphism (SNP) technology has matured sufficiently so that the throughput of molecular marker-based genotyping is able to keep pace with the numbers of plants that breeders routinely handle in the field. We explore the extent to which the genomics revolution might generate a change in the conventional breeding paradigm, which has operated with such success for the best part of the 20th century, and identify the need for a substantial reduction in assay price before MAS is likely to make more than a marginal impact on present practice. PMID- 12573854 TI - Two approaches for metabolic pathway analysis? AB - Metabolic pathway analysis is becoming increasingly important for assessing inherent network properties in (reconstructed) biochemical reaction networks. Of the two most promising concepts for pathway analysis, one relies on elementary flux modes and the other on extreme pathways. These concepts are closely related because extreme pathways are a subset of elementary modes. Here, the common features, differences and applicability of these concepts are discussed. Assessing metabolic systems by the set of extreme pathways can, in general, give misleading results owing to the exclusion of possibly important routes. However, in certain network topologies, the sets of elementary modes and extreme pathways coincide. This is quite often the case in realistic applications. In our opinion, the unification of both approaches into one common framework for metabolic pathway analysis is necessary and achievable. PMID- 12573855 TI - Radiopharmaceuticals: new antimicrobial agents. AB - Small antimicrobial peptides are good candidates for new antimicrobial agents. A scintigraphic approach to studying the pharmacokinetics of antimicrobial peptides in animals has been developed. The peptides were safely and reproducibly labelled with technetium-99m and, after intravenous injection of the radiolabelled peptides into infected animals, scintigraphy allowed real-time quantification of the peptide in the various body compartments. Antimicrobial peptides rapidly accumulated at sites of infection but not at sites of sterile inflammation, indicating that radiolabelled antimicrobial peptides could be used in detection of infection. These radiopharmaceuticals enabled the efficacy of antibacterial therapy in animals to be monitored. The scintigraphic approach provides a useful method for investigating the pharmacokinetics of small peptides in animals. PMID- 12573856 TI - LNA: a versatile tool for therapeutics and genomics. AB - Locked nucleic acid (LNA) is a nucleic acid analogue that displays unprecedented hybridization affinity towards complementary DNA and RNA. Structural studies have shown LNA to be an RNA mimic, fitting seamlessly into an A-type duplex geometry. Several reports have revealed LNA as a most promising molecule for the development of oligonucleotide-based therapeutics. For example, Tat-dependent transcription and telomerase activity have been efficiently suppressed by LNA oligomers, and efficient cleavage of highly structured RNA has been achieved using LNA-modified DNAzymes ('LNAzyme'). Furthermore, convincing examples of the application of LNA to nucleic acid diagnostics have been reported, including high capturing efficiencies and unambiguous scoring of single-nucleotide polymorphisms. PMID- 12573858 TI - Sonobioreactors: using ultrasound for enhanced microbial productivity. AB - Enhanced metabolic productivity of microbial, plant and animal cells in bioreactors can greatly improve the economics of biotechnology processes. Ultrasound is one method of intensifying the performance of live biocatalysts. Ultrasonication is generally associated with damage to cells but evidence is emerging for beneficial effects of controlled sonication on conversions catalyzed by live cells. This review focuses on the productivity enhancing effects of ultrasound on live biological systems and the design considerations for sonobioreactors required for ultrasound-enhanced biocatalysis. PMID- 12573857 TI - Global organellar proteomics. AB - Cataloging the proteomes of single-celled microorganisms, cells, biological fluids, tissue and whole organisms is being undertaken at a rapid pace as advances are made in protein and peptide separation, detection and identification. For metazoans, subcellular organelles represent attractive targets for global proteome analysis because they represent discrete functional units, their complexity in protein composition is reduced relative to whole cells and, when abundant cytoskeletal proteins are removed, lower abundance proteins specific to the organelle are revealed. Here, we review recent literature on the global analysis of subcellular organelles and briefly discuss how that information is being used to elucidate basic biological processes that range from cellular signaling pathways through protein-protein interactions to differential expression of proteins in response to external stimuli. We assess the relative merits of the different methods used and discuss issues and future directions in the field. PMID- 12573859 TI - 226Ra, 40K and 7Be activity concentrations in plants in the environment of Kaiga, India. AB - Leaves, stem and bark samples from several plant species were collected from tropical forest of Kaiga, in the west coast of India where two nuclear power reactors of 220 MW each have just been commissioned and another two are under construction, and analysed for their (226)Ra, (40)K and (7)Be concentrations. The activities of (226)Ra and (40)K in plants were found to vary in the range BDL 13.2 and 12.0-797.3 Bq kg(-1), respectively. Plants show significant (7)Be activity in leaves, the activity varies in 72.5-1,060.8 Bq kg(-1). Stem and bark of plants show higher levels of (226)Ra and (40)K when compared to leaves. Soil to-plant transfer factor for (226)Ra and (40)K were found to vary in the range BDL-0.37 and 0.09-5.61, respectively for different plants. The concentration of (226)Ra and (40)K in leaves depends on the age of the leaves. PMID- 12573860 TI - Sediment accumulation rate and radiological characterisation of the sediment of Palmones River estuary (southern of Spain). AB - Chemical analyses and radioecological methods were combined in order to estimate the sediment accumulation rate in the upper 20 cm depth of the Palmones River estuary. Organic matter, total carbon, C:N and (137)Cs vertical profiles showed changes at 13 cm depth. These changes could be associated with the decrease in river input since 1987 when a dam situated in the upper part of the estuary started to store water. Using 1987 as reference to date the sediment, accumulation rate was 1.2 cm yr(-1). As alternative method, two layer model of (210)Pb(xs) vertical distribution showed a sedimentation rate of 0.7 cm yr(-1) with a surface mixing layer of 7 cm thickness. The high ammonium, potassium and sodium content in pore water and the strong correlation between (137)Cs activities and organic matter in dry sediment suggests that (137)Cs (the only anthropogenic product detected) is mainly accumulated in the estuary associated with the particulate organic material from the catchment area. PMID- 12573861 TI - Spatial variability of fallout-90Sr in soil and vegetation of an alpine pasture. AB - According to the soil-to-plant transfer concept generally used in dose assessment modeling, the plant uptake of a radionuclide should depend linearly on its concentration in the soil. In order to validate this concept for (90)Sr in a semi natural ecosystem, plant and soil samples were taken at 100 plots of a 100 x 100 m(2) area within an alpine pasture near Berchtesgaden, Germany. At three plots, the vertical distribution of (90)Sr in the soil was determined in addition. A statistically significant correlation between the soil and plant concentration of (90)Sr was not detectable (Spearman correlation coefficient R=-0.116, p>0.05) within the range of the Sr-concentration covered (15-548 Bq kg(-1) dry soil and 17-253 Bq kg(-1) dry plant material). Thus, the prerequisite of the soil-to-plant transfer concept was not fulfilled for (90)Sr at this site. Organic carbon and total nitrogen were also determined in the soil samples. Both elements were highly correlated (R=0.912, p<0.001), their ratio being C/N=10.9+/-0.7. While C was positively correlated with the (90)Sr concentrations in the soil (R=0.342, p<0.001), negative correlations were observed for the plant concentrations (R= 0.286, p<0.01) and the concentration ratios (R=-0.444, p<0.001) of (90)Sr. These results are compared with those recently obtained for (137)Cs by Bunzl et al. (J Environ Radioactiv 48 (2000) 145). PMID- 12573862 TI - 110mAg root and foliar uptake in vegetables and its migration in soil. AB - 110mAg, as a radionuclide of corrosion products in water-cooled nuclear reactors, was detected in the liquid effluents of Guangdong Daya Bay Nuclear Power Station (GNPS) of Daya Bay under normal operation conditions. Experiments on a simulated terrestrial agricultural ecosystem were carried out using the pot experiment approach. The most common plants in Hong Kong and the South China vegetable gardens such as lettuce, Chinese spinach, kale, carrot, pepper, eggplant, bean, flowering cabbage, celery, European onion and cucumber were selected for (110m)Ag root and foliar uptake tests. The results show that carrot, kale and flowering cabbage have the greatest values of soil to plant transfer factor among the vegetables, while(110m)Ag can be transferred to Chinese spinach via foliar uptake. Flowering cabbage, the most popular leafy vegetable locally, could be used as a biomonitor for the radioisotope contamination in vegetables. Soil column and adsorption tests were also carried out to study the leaching ability and distribution coefficient (K(d)) of (110m)Ag in the soil. The results show that most of the radionuclide was adsorbed in the top 1 cm of soil regardless of the pH value. The K(d) was also determined. PMID- 12573863 TI - Ventomod: a dynamic model for leaf to fruit transfer of radionuclides in processing tomato plants (Lycopersicon esculentum Mill.) following a direct contamination event. AB - This paper presents results on the calibration and validation of a model (Ventomod) for leaf to fruit transfer of (134)Cs, (85)Sr and (65)Zn in processing tomato plants after leaf contamination. Several models (e.g. FARMLAND) that deal specifically with the transfer of radionuclides to fruits are adaptations of models that were developed for agricultural crops such as leafy green vegetables. "Ventomod" represents a dynamic evaluation model exclusively built for the short term behaviour of radionuclide depositions. It forecasts the level of radionuclide contamination in ripe processing tomato fruits following an accidental radionuclide release into the atmosphere. A validation of the developed model by data sets from an independent experiment showed that the model successfully reproduced the observed radionuclide distribution and dynamics in tomato fruits. The level of uncertainty was within the normal range of similar assessment models. For a more general use of this model further testing with independent data sets from experiments obtained under different environmental conditions and data from other horticulturally important plant species would be desirable. PMID- 12573864 TI - Prediction of the fate of radioactive material in the South Pacific Ocean using a global high-resolution ocean model. AB - We investigate the release of radioactive contaminants from Moruroa Atoll in a global high-resolution off-line model. The spread of tracer is studied in a series of simulations with varying release depths and time-scales, and into ocean velocity fields corresponding to long-term annual mean, seasonal, and interannually varying scenarios. In the instantaneous surface release scenarios we find that the incorporation of a seasonal cycle greatly influences tracer advection, with maximum concentrations still found within the French Polynesia region after 10 years. In contrast, the maximum trace is located in the southeast Pacific when long-term annual mean fields are used. This emphasizes the importance of the seasonal cycle in models of pollution dispersion on large scales. We further find that during an El Nino/Southern Oscillation (ENSO) event reduced currents in the region of Moruroa Atoll result in increased concentrations of radioactive material in French Polynesia, as direct flushing from the source is reduced. In terms of the sensitivity to tracer release time rates, we find that a gradual input results in maximum concentrations in the near vicinity of French Polynesia. This contrasts the instantaneous-release scenarios, which see maximum concentrations and tracer spread across much of the South Pacific Ocean. For example, in as little as seven years radioactive contamination can reach the east coast of Australia diluted by only a factor of 1,000 of the initial concentration. A comparison of results is made with previous studies. Overall, we find much higher concentrations of radionuclides in the South Pacific than has previously been predicted using coarser-resolution models. PMID- 12573865 TI - Regional dependence of urinary uranium baseline levels in non-exposed subjects with particular reference to volunteers from Northern Italy. AB - Knowledge of the level of natural uranium (U) in the human body is fundamental in order to estimate the potentially hazardous incorporation in accidentally exposed subjects. A constant monitoring of exposed workers needs reliable reference baseline values, which can be determined by measuring the U concentration in urine. ICPMS has proven to be a fast, reliable and highly sensitive technique for this purpose. Non-uniformity in the distribution of U levels in various regions and differences in dietary habits account for the significant regional variations of U concentration in urine in non-exposed subjects. In this paper, the determination of daily uranium urinary excretion levels in a group of 12 non exposed subjects from Northern Italy is presented and compared to data present in the published literature and to values obtained in a larger group of German volunteers. The urinary U output values observed in the Italian subset are generally higher than the corresponding levels measured in other groups. This could be the result of a higher intake of U from liquids, as assessed by the determination of U concentration in drinking waters. PMID- 12573867 TI - The contribution of the MPTP-treated primate model to the development of new treatment strategies for Parkinson's disease. AB - Current research into Parkinson's disease (PD) is directed at developing novel agents and strategies for improved symptomatic management. The aim of this research is to provide effective and maintained symptom control throughout the course of the disease without loss of efficacy and without priming the basal ganglia for the onset of dyskinesia. To achieve these objectives, it is important to have relevant animal models of PD in which new pharmacological agents and treatment strategies can be assessed prior to clinical assessment. At present, the most effective experimental model of PD is the methyl phenyl tetrahydropyridine (MPTP)-treated primate. Primates treated with MPTP develop motor disturbances resembling those seen in idiopathic PD, including bradykinesia, rigidity and postural abnormalities. In addition, MPTP-treated primates are responsive to all commonly used antiparkinsonian agents and display treatment-associated motor complications such as dyskinesia, wearing-off and on off, which occur during the long-term treatment of the illness. This review examines how studies conducted in MPTP-treated primates have contributed to the development of dopaminergic therapies. There is now accumulating evidence that the pulsatile manner in which short-acting agents stimulate striatal dopamine receptors is a key contributing factor to the priming of the basal ganglia for dyskinesia induction. It has been suggested that providing more continuous stimulation of dopamine receptors will avoid the development of motor complications, particularly dyskinesia. So far, the actions of all commonly used antiparkinsonian drugs assessed in MPTP-treated primates have proved to be highly predictive of drug action in PD. These primate studies have demonstrated that long-acting dopamine agonists and levodopa given in combination with a catechol-O methyl transferase (COMT) inhibitor (to increase its relatively short half-life), induce significantly less dyskinesia than occurs with standard levodopa therapy. PMID- 12573868 TI - The effect of apomorphine administration on smooth pursuit ocular movements in early Parkinsonian patients. AB - Electrooculography (EOG) recordings in 21 L-DOPA-naive patients suffering from Parkinson's disease (PD) were made before and after apomorphine subcutaneous administration (ASA). The effect of apomorphine on smooth pursuit eye movements (SPEM) was studied. Age-matched healthy subjects, who underwent SPEM recordings without the ASA procedure, were examined in order to compare baseline SPEM. EOG recordings were used to compare the patient group and the control group, and to compare the SPEM before and after ASA within the patient group. Significant differences in SPEM were found between both groups, as well as in the SPEM before and after ASA. The theory that SPEM is disturbed in early PD patients was confirmed. The dopaminergic control of horizontal SPEM is supposed. PMID- 12573869 TI - Effects of peripheral and central catechol-O-methyltransferase inhibition on striatal extracellular levels of dopamine: a microdialysis study in freely moving rats. AB - Tolcapone is a mixed (peripheral and central) catechol-O-methyltransferase (COMT) inhibitor, whereas entacapone is a preferential peripheral COMT inhibitor. Both drugs are able to decrease the peripheral conversion of L-DOPA into 3-O-methyl DOPA and thereby increase plasma and cerebral levels of L-DOPA, the precursor of dopamine (DA). Tolcapone may also impair the extraneuronal catabolism of DA by inhibiting COMT activity in the brain. To evaluate the role played by peripheral and central COMT inhibition, we compared the effects of tolcapone and entacapone on COMT activity in peripheral tissues, and on striatal extracellular levels of L DOPA and DA in rats. Tolcapone and entacapone, at the dose of 15 mg/kg p.o., were almost equally effective in inhibiting COMT activity in duodenum and liver. Tolcapone decreased striatal extracellular levels of homovanillic acid (HVA), thus confirming its central COMT inhibitory effect, whereas entacapone did not alter HVA efflux. Following L-DOPA/benserazide administration (50/15 mg/kg p.o.), both COMT inhibitors significantly increased striatal levels of L-DOPA and DA compared with saline. The levels of L-DOPA were similar after treatment with either COMT inhibitors, whereas the increase in DA output was significantly greater in rats given tolcapone compared to those given entacapone. We conclude that the blockade of central DA catabolism by tolcapone contributes to the greater increase in striatal DA levels achieved with this drug. PMID- 12573870 TI - Clinical-electrophysiological correlation of tremor and myoclonus in a kindred with the N279K tau mutation. AB - We used electrophysiological methods to study the hyperkinetic movement disorders in a pallido-ponto-nigral degeneration (PPND) family, which harbors the N279K tau gene mutation. Our purpose was to: (1). characterize the tremor patterns, (2). characterize the myoclonus physiology, (3). determine whether electrophysiology can detect abnormalities in asymptomatic cases. In PPND, we found that the activation tremors correlated with a semi-rhythmic 6-10 Hz electromyography (EMG) pattern, and the rest tremors showed a reciprocal 4-6 Hz pattern. At least two different myoclonus physiology patterns exist in PPND, most notably contrasted by the presence or absence of a demonstrable electroencephalography (EEG) correlate. Electrophysiology yielded remarkable findings in those asymptomatic at-risk individuals that carried the N279K tau mutation. PMID- 12573871 TI - Bilateral subthalamic nucleus stimulation after bilateral pallidotomies in a patient with advanced Parkinson's disease. AB - A 54-year-old man with advanced Parkinson's disease (PD) presented to our institution in early 2000. He had undergone a right pallidotomy in 1994, a left pallidotomy in 1996, and bilateral subthalamic nucleus (STN) electrode implants in 1999. The patient had cervical myelopathy for which he had undergone neck surgery in 1998. We used the Unified Parkinson's Disease Rating Scale (UPDRS) to evaluate motor performance in four states: combinations of stimulation OFF or ON and medication OFF or ON. There was no significant change in motor UPDRS scores with STN stimulation or with medications. Multiple attempts to optimize stimulation parameters and medication dosages did not result in significant and sustained improvement in activities of daily living or motor performance. To our knowledge, this is the first reported case of bilateral STN stimulation after bilateral pallidotomies. The presence of cervical myelopathy and the limited response to anti-Parkinson medications in this patient underscores the importance of patient selection for functional neurosurgery in PD. PMID- 12573872 TI - Economic burden and quality of life impairment increase with severity of PD. AB - PURPOSE: To define the interrelationship between cost-of-illness, quality of life (QoL) and Parkinson's disease (PD) severity in a common patient management setting in Finland.Scope. Two hundred and sixty consecutive outpatients with idiopathic PD participated. UPDRS, motor fluctuations, QoL, and the use of health care resources were measured. Direct and indirect costs were calculated. CONCLUSIONS: There is a strong relationship between QoL or cost-of-illness on the one hand, and severity of PD on the other. Treatment policies capable of reducing or delaying motor fluctuations would be expected to increase QoL and reduce some of the economic burden of PD. PMID- 12573873 TI - Freezing of gait in multiple system atrophy (MSA). AB - BACKGROUND: Freezing of gait (FOG) is a mysterious symptom, observed in different parkinsonian syndromes, but considered to be rare in multiple system atrophy (MSA). OBJECTIVE: To assess the frequency of FOG in patients with MSA. METHOD: We studied the presence of FOG in 28 patients with clinical diagnosis of MSA. 21 patients had probable MSA and 7 had possible MSA. The clinical diagnosis was based on neurological examination performed by at least two experienced movement disorders specialists as well as on the results of ancillary examinations and the course of the disease. In 22 patients MSA was predominated by parkinsonism (MSA P) and in 6 by cerebellar signs (MSA-C). The patients' mean age was 66.8+/-10.3 years, mean disease duration 6.4+/-4.0 years and mean worst Hoehn and Yahr (H&Y) stage was 3.6+/-0.6. Presence and severity of FOG was assessed during the last office visit, by the recently validated FOG questionnaire (FOG-Q), with a maximal score of 24, while patients that received at least one point in the last four questions were classified as having FOG. Severity of FOG was determined by the sum of these last four out of six questions (maximal score is 16). FOG-Q total score reflected general function and walking ability disturbances, caused by FOG. The comparison between groups of patients was performed by way of the Mann Whitney two-sample test and chi-square or Fisher's exact tests. Correlations between various parameters were calculated using Spearman's correlation coefficient. RESULTS: Twenty-one patients were able to walk and 7 were bedridden at the time of the study. FOG appeared in a total of 75% of all MSA patients (in 82% of patients with MSA-P and in 50% patients with MSA-C). In the MSA-P group disease duration was about the same among 'freezers' and 'non-freezers', while among the MSA-C patients it was significantly shorter in the non-freezers. Mean score of the freezing subdivision of the FOG-Q was 8.2+/-5.1 for MSA-P group and 4.5+/-5.1 for MSA-C one. Mean FOG-Q total score was 9.1+/-4.0 and 6.2+/-4.6 (p>0.05) for MSA-P and MSA-C patients, respectively. CONCLUSION: Freezing of gait is a common symptom in MSA, both in MSA-P and MSA-C. PMID- 12573874 TI - Neuroleptic malignant syndrome-like, or--dopaminergic malignant syndrome--due to levodopa therapy withdrawal. Clinical features in 11 patients. AB - A clinical picture named neuroleptic malignant-like syndrome has been described in patients with Parkinson's disease (PD) who suddenly stop their L-dopa treatment. The sudden withdrawal of the drug is deemed to lead to an acute deficiency stage in a patient who has an iatrogenic increase of dopaminergic transmission. We present a series of 11 patients with PD with an average age of 72.09 years, a mean disease duration of 9.45 years who developed this problem after a 92.72 h latency period. If patients with PD develop severe rigidity, stupor and hyperthermia, L-dopa withdrawal should be suspected and the dopaminergic drug restarted as soon as possible. PMID- 12573875 TI - Are electrophysiological autonomic tests useful in the assessment of dysautonomia in Parkinson's disease? AB - To assess the autonomic system in Parkinson's disease (PD), the sympathetic skin response (SSR) and the R-R interval variation (RRIV) tests were studied in 26 PD patients and in 24 healthy controls. The aim of the study was to evaluate the sympathetic and parasympathetic system function in PD, to define the pattern of autonomic abnormalities found in SSR and RRIV in parkinsonian patients as well as to analyze the usefulness of both tests in paraclinical assessment of the dysautonomia, compared with clinical symptoms and signs of the autonomic nervous system involvement. The corrrelations between both autonomic tests results were also studied. In PD patients SSR test was abnormal in about 35% and RRIV was abnormal in about 54% of patients. SSR and RRIV were both abnormal in about 27% of PD patients whereas at least one of electrophysiological autonomic tests was abnormal in about 62% of PD patients. Clinical and paraclinical signs of dysautonomia occurred in a similar proportion of patients (i.e. in about 62%). A weak correlation was found between the latency of SSR from upper limbs and the value of RRIV during deep breathing (p=0.063). Our results show that SSR and RRIV are non-invasive paraclinical electrophysiological tests that confirm clinical dysautonomia in PD and can supplement the clinical differentiation of Parkinsonian syndromes. PMID- 12573876 TI - Carotid sinus nerve chemosensory response to dopamine and acetylcholine in catecholamine depleted cats. AB - The aim of this study was to determine the role of endogenous dopamine (DA) and the combined effect of DA and acetylcholine (ACh) on the carotid sinus nerve chemosensory discharge (CSND). CSND was measured in vivo in 6 control cats and 6 cats pre-treated with reserpine and alpha-methyl-paratyrosine [catecholamine depleted group: CAD] during infusions of DA and DA+ACh. In normoxia, CSND was similar between CAD's and controls. DA induced CSND depression was transient in controls but sustained in CAD's. Addition of ACh increased CSND in both groups. In hypoxia (8% O(2) in N(2)), the dynamic CSND response was slowed by DA in CAD's but not controls. Addition of ACh increased this response in both groups. Neither DA nor DA+ACh altered the steady state hypoxic CSND in either group. It is concluded that endogenous DA is important in expressing the dynamic characteristics of both the response to exogenous DA and the response to hypoxia under constant DA infusion. The study also confirms the opposing effects of exogenous DA and ACh on the normoxic CSND. PMID- 12573877 TI - Effect of obesity and/or sleep apnea on chemosensitivity: differences between men and women. AB - The purpose of this study was to identify if obesity or obstructive sleep apnea (OSA) influence hypercapnic response (HCR) and hypoxic response (HR) taking into account differences between gender. Data are expressed in % predicted based on reference values of our laboratory ('Respir. Physiol. 113 (1998) 157'). Obese women without OSA (n=117, body mass index (BMI) 43+/-8 kg/m(2)) demonstrated an increased ventilatory (VE) and occlusion pressure (P(0.1)) HCR and HR slope: VE HCR 113 (NS), VE HCR/vital capacity (VC) 126 (P<0.05), P(0.1) HCR 130 (P<0.05), VE HR 136 (P<0.05), VE HR/VC 154 (P<0.001) and P(0.1) HR 210 (P<0.001) % predicted. Obese women with OSA (n=34, BMI 42+/-9 kg/m(2)) presented similar increased values for HCR and even more increased values for HR than obese women without OSA matched for age, height and BMI: VE HR 155 (P=NS), VE HR/VC 205 (P<0.05) and P(0.1) HR 273 (P<0.05) % predicted. In obese men (without or with OSA) HCR and HR values were similar to our reference values. PMID- 12573878 TI - Modeling of the expiratory flow pattern of spontaneously breathing cats. AB - A mathematical model was developed describing the entire expiratory flow pattern during spontaneous, tidal breathing in the absence of expiratory muscle activity. It provides estimates for the time constants of the respiratory system (tau RS(model)) and of the decay of continuing inspiratory muscle activity in early expiration (tau mus(model)). In ten anesthetized, tracheostomized cats flow, tracheal pressure and diaphragmatic EMG were measured during normal expirations and expirations with four different added resistances. No significant differences were found between tau RS(model) (0.21-0.49 sec) obtained by fitting the model to the flow data and tau RS obtained from the straight part of the expiratory flow volume curve. tau mus(model) (0.050-0.052 sec) was comparable to similar time constants obtained from the integrated diaphragmatic EMG or from end-inspiratory, tracheal occlusion pressure. Fitted peak flow and time to peak tidal expiratory flow were not significantly different from those measured. In conclusion, for spontaneously breathing, anesthetized cats our model provides a close fit of the expiratory flow and parameter estimates were comparable with independently measured values. PMID- 12573879 TI - The indirect effect of Tityus discrepans on rabbit pulmonary vasculature. AB - Serum (IS) was obtained 0.5, 2, 4 or 6 h after inoculating s.c. six rabbits (approximately 2 kg) in each time period with 1 mg/kg of Tityus discrepans (Td) venom; the control was serum obtained from four rabbits 4 h after injecting them 1 ml s.c. of 0.9% NaCl. IS produced a transient (<25 min) rise in pulmonary artery pressure of isolated and perfused rabbit lungs, other lung parameters were not altered. We found that both scorpion venom and IS produced a approximately 50% transient increase of transendothelial electric resistance in cultured tissue human umbilical cord vein. Neither venom nor IS changed the transepithelial electrical resistance of tissue cultured human airway epithelia. The experiments suggest that humoral factors contained in the inoculated serum modify vascular endothelium in a much more effective manner than the venom by itself. These experiments also make it unlikely that vascular endothelium is the source of the humoral factors contained in inflammatory serum. PMID- 12573880 TI - Morphometry and allometry of the postnatal lung development in the quokka wallaby (Setonix brachyurus): a light microscopic study. AB - The postnatally developing lungs of the quokka wallaby, Setonix brachyurus, were investigated macroscopically and by light microscopic morphometry. Lung, parenchymal and non-parenchymal volumes as well as the components of the latter two were analysed by regression analysis. The lungs comprised a single undivided left lung and a right lung with an adherent accessory lobe. Septal tissue growth was most remarkable in the canalicular and saccular stages. Between mid canalicular stage and the saccular stage, the lung volume increased 2-fold, mainly due to airspace expansion, coupled with septal tissue thinning. The non parenchymal vascular volume increase accelerated in the successive developmental stages while the airway and connective tissue volumes progressed in a decreasing order, being highest in the canalicular and saccular stages and lowest in the alveolar stage. Growth and remodelling of the alveolar septa occurred simultaneously with airspace subdivision. Airspace expansion accelerated during the stage of microvascular maturation, when most other parameters showed the least rate of increase. PMID- 12573881 TI - Open-flow plethysmography with pressure-decay compensation. AB - Whole-body plethysmography is widely used to measure ventilation in awake, unrestrained animals. However, the explicit solution for volumetric analysis of the plethysmograph signal depends upon a closed system, which limits experimental design. Although often used, open-flow plethysmography is complicated by the time decay of pressure signals generated in the open chamber (e.g. equivalent volume displacements will yield different pressure pulse magnitudes depending upon the rate of application, dP/dt). This problem may be alleviated by first characterizing the time rate of pressure-decay, dP(k)/dt, as a function of pressure magnitude, P, in the plethysmograph, dP(k(P))/dt. Then for each point P(t) in the original signal, subtract the corresponding dP(k(P))(t)/dt from each dP(t)/dt of the original signal to determine the decay-compensated derivative for that point, dP*(t)/dt, and then numerically integrate dP*(t)/dt to generate a pressure-decay compensated signal. The result is a 'virtual closed plethysmograph' trace that enables confident quantitative determination of ventilatory events and volumes with the full advantage of an open-flow plethysmograph. PMID- 12573882 TI - Sustained hypoxia promotes hyperactive response of carotid body in the cat. AB - Carotid body chemosensory activities were measured before and after 0.2, 5,6 and 7 h of sustained isocapnic (PaCO(2) approximately equal to 30 Torr) hypoxia (PaO(2) approximately equal to 43 Torr) in the cats (n=7). The activity increased from 5.4 impsec(-1) at 0.2 h to about 13 impsec(-1) at 7 h. This increase in chemosensory activities were due to both an augmented sensitivity and to a long term facilitation and not due to arterial [H(+)] changes. PMID- 12573883 TI - Concept of a PACS and imaging informatics-based server for radiation therapy. AB - Radiation Therapy (RT) is an image-based treatment. It requires images from projection X-rays, computed tomography, magnetic resonance, positron emission tomography, Linear Accelerator for tumor localization, treatment planning and verification of treatment plans. During the treatment process, patient's images are transmitted to every necessary station in the RT department. However, images of the same patient are generally scattered and there is no permanent home base for them due to the nature and traditional organization of the RT department. The advance in diagnostic picture archiving and communication system and the establishment of RT DICOM Standard provide an opportunity to define and design an RT server as a means to organize RT images and related data. This paper describes the RT workflow and the concept of the DICOM RT server. An example of RT treatment of nasopharyngeal carcinoma based on the RT server concept is given. PMID- 12573884 TI - T2 dark blood MRA for renal artery stenosis detection: preliminary observations. AB - OBJECTIVE: This study evaluated the ability of a fast spin echo T2 weighted dark blood sequence to characterize significant (>50%) renal artery stenosis compared to conventional angiography. METHODS: Sixteen patients underwent conventional catheter angiography for either renal artery stenosis evaluation or as potential renal donors. Each patient then had an MR study of the renal arteries and kidneys with fast spin echo T2 weighted MR (TR 4000, TE 102, 8 echo train length) on a Superconducting 1.5T Magnet. Results were compared with angiography and inter and intra observer statistics were calculated. RESULTS: A total of 36 renal arteries were imaged in 32 kidneys with 12 stenoses >50%. Fast spin echo T2 weighted MR is 94% accurate (95%CI: 87-100%) in detection of significant renal artery stenosis. Dark blood MRA (DBMRA) is 96% sensitive (95%CI: 89-100), 92% specific, with a predictive value positive of 96% for classifying real arteries as normal or significantly stenosed. Inter and intra observer statistics demonstrate good to excellent agreement in renal artery classification (kappa>0.60). CONCLUSION: DBMRA may be a useful adjunct to renal MR evaluation in hypertension. SUMMARY: A total of 36 renal arteries were imaged in 32 kidneys with 12 stenoses >50%. Fast spin echo T2 weighted MR is 94% accurate (95%CI: 87-100%) in detection of significant renal artery stenosis. PMID- 12573885 TI - Diffusion MRI findings in Wilson's disease. AB - Six patients having Wilson's disease were studied with diffusion MRI in order to characterize cerebral lesions. Diffusion MRI was obtained using the spin-echo, echo-planar sequence with a gradient strength of 30 mT/m. The trace protocol was used in the axial imaging plane. Heavily diffusion-weighted (b=1000s/mm(2)) images, and the ADC (apparent diffusion coefficient) values from automatically generated ADC maps were studied. The ADC values of the normal brain parenchyma were available in 17 age-matched cases for comparison (ADC values, 0.85+/-0.11 x 10(-3)mm(2)/s). In Wilson's disease two distinct diffusion MRI patterns were observed by quantitative evaluations of the ADC maps; cytotoxic edema-like (ADC values, 0.52+/-0.03 x 10(-3)mm(2)/s), and vasogenic edema-like (ADC values, 1.42+/-0.17 x 10(-3)mm(2)/s) patterns. Diffusion imaging appears to be a promising sequence to evaluate the changes in the brain tissue in Wilson's disease at least by revealing two different patterns. PMID- 12573886 TI - Proton MR spectroscopy demonstration of taurine peaks in megalencephalic leukoencephalopathy with cysts. AB - Taurine is an amino acid. It has roles in neurotransmitter activities, facilitates synaptic connections, and possesses cyto-protective properties. Normally, it is invisible on proton MR spectroscopy acquired at low field strengths (i.e. 1.5T). In a patient with megalencephalic leukoencephalopathy with cysts (MLC), distinct taurine peaks centered at 3.35ppm were found on hybrid-CSI spectroscopy obtained at 1.5T with a long TE value (i.e. TE=135ms). This suggested increased concentrations of this metabolite in MLC. In this leukoencephalopathy taurine may be linked to a metabolic disturbance related with neurotransmitters associated with a disturbance in synaptic connections, and cyto protective mechanisms. Further spectroscopic and clinical studies are required to evaluate this observation. PMID- 12573887 TI - CT and MRI in malignant external otitis: a report of four cases. AB - Malignant external otitis (MEO) is a severe infection of the external auditory canal and skull base which most often affects elderly patients with diabetes mellitus. Early diagnosis and aggressive treatment are important in terms of reducing risks of dissemination. We report four cases of MEO in patients with diabetes mellitus who were observed with computed tomography (CT) and/or magnetic resonance imaging (MRI). MRI was superior to CT in patients with MEO in terms of estimating the anatomic extent of the disease, but it cannot be used for monitoring therapy. PMID- 12573888 TI - Clinical usefulness of the cardiac multi-detector-row CT. AB - Along with improvement of the temporal and special resolution, multi-detector-row CT has become able to generate artifact-less heart images. We discuss the potential benefits of the newly developed cardiac application, demonstrating informative cases.Two- and three-dimensional (2D and 3D) cardiac images were produced throughout one cardiac cycle. By paging 2D and 3D images in cardiac cycle order, animated movies were generated. Cardiac imaging with multi-detector row CT, acquired during a single breath hold, provides information about: (1) clear morphology of heart chambers and myocardium, (2) wall motion and systolic thickening, (3) myocardial perfusion, (4) volume assessment, and (5) coronary anatomy and pathophysiology. PMID- 12573889 TI - Performance evaluation of principal component analysis in dynamic FDG-PET studies of recurrent colorectal cancer. AB - Performance evaluation of principal component analysis (PCA) of dynamic F-18-FDG PET studies of patients with recurrent colorectal cancer. Principal component images (PCI) of 17 iteratively reconstructed data sets were visually and quantitatively evaluated. The F-18-FDG compartment model parameters were estimated using polynomial regression. All structures were present in PCI1. PCI2 was correlated with the vascular component and PCI3 with the tumor. The vessel density in the tumor was estimated with a correlation coefficient equal to 0.834. PCA supports the visual interpretation of dynamic F-18-FDG-PET studies, facilitates the application of compartment modeling and is a promising quantification technique. PMID- 12573890 TI - Experimental study of stochastic noise propagation in SPECT images reconstructed using the conjugate gradient algorithm. AB - Thanks to an experimental study based on simulated and physical phantoms, the propagation of the stochastic noise in slices reconstructed using the conjugate gradient algorithm has been analysed versus iterations. After a first increase corresponding to the reconstruction of the signal, the noise stabilises before increasing linearly with iterations. The level of the plateau as well as the slope of the subsequent linear increase depends on the noise in the projection data. PMID- 12573891 TI - Towards a computer-aided diagnosis system for pigmented skin lesions. AB - This paper presents a computer-aided diagnosis system for pigmented skin lesions, with solutions for the lesion boundary detection and for the quantification of the degree of symmetry. Lesion detection results were validated by expert dermatologists, who also provided hand-drawn boundaries of the lesions. These reference boundaries were not used as a gold standard, but were allowed to statistically determine the accuracy of the boundaries provided by computerized techniques. We could show that the dermatologists were not able to reproduce their results, and that the boundaries of any expert taken alone showed higher divergence from those of the set of remaining experts than the automatic techniques we developed. Feature extraction is restricted in this paper to the quantification of degree of symmetry, even though it is clear that many other features will be necessary for a complete diagnosis system. The symmetry quantification step provides a six-dimensional feature vector that can be used to classify pigmented skin lesions as being benign or malignant. We demonstrate that our scheme outperforms methods based on the principal component decomposition, which is widely used for this kind of application. PMID- 12573892 TI - A novel approach for distortion correction for X-ray image intensifiers. AB - Applications of X-ray image intensifiers in medical imaging, include the use of fluoroscopic projection images to generate three-dimensional tomographic reconstructions. Unfortunately, the inherent distortions on the acquired projections deteriorate the quality of the reconstructed tomograms. Distortion correction can be performed using algorithms that can be classified as global or local according to the method used, both having specific advantages and disadvantages. In this work, a novel approach for distortion correction is proposed which, by combining both global and local correction methods allows good image quality in relatively acceptable time. Correction parameters were obtained using a calibration phantom specially designed for this purpose. PMID- 12573893 TI - Leukoencephalopathy with patchy lesions, temporal cystic degeneration, and normal head size. AB - A 5 yr old girl is reported with patchy white matter lesions, cystic degeneration of the temporal tips, normal cortex, and normal cerebellum, documented by FLAIR, and diffusion-weighted MRI. This condition differed from the entity megalencephalic leukoencephalopathy with cysts by lack of diffuse involvement of the white matter and cortical thinning, by presence of a normal head size, and existence of a non-progressive, severe clinical condition. Occurrence of cystic degeneration in the temporal tips distinguished the condition from other leukoencephalopathies. This leukoencephalopathy appears to be a new distinct entity. PMID- 12573894 TI - Ethical perspectives for public and environmental health: fostering autonomy and the right to know. AB - In this paper we develop an ethical perspective for public and environmental health practice in consideration of the "right to know" by contrasting consequential and deontological perspectives with relational ethics grounded in the concept of fostering autonomy. From the consequential perspective, disclosure of public and environmental health risks to the public depends on the expected or possible consequences. We discuss three major concerns with this perspective: respect for persons, justice, and ignorance. From a deontological perspective, the "right to know" means that there is a "duty" to communicate about all public health risks and consideration of the principles of prevention, precaution, and environmental justice. Relational ethics develops from consideration of a mutual limitation of the traditional perspectives. Relational ethics is grounded in the relationship between the public and public/environmental health providers. In this paper we develop a model for this relationship, which we call "fostering autonomy through mutually respectful relationships." Fostering autonomy is both an end in public health practice and a means to promote the principles of prevention, precaution, and environmental justice. We discuss these principles as they relate to practical issues of major disasters and contaminants in food, such as DDT, toxaphene, chlordane, and mercury. PMID- 12573895 TI - Mechanisms of phthalate ester toxicity in the female reproductive system. AB - Phthalates are high-production-volume synthetic chemicals with ubiquitous human exposures because of their use in plastics and other common consumer products. Recent epidemiologic evidence suggests that women have a unique exposure profile to phthalates, which raises concern about the potential health hazards posed by such exposures. Research in our laboratory examines how phthalates interact with the female reproductive system in animal models to provide insights into the potential health effects of these chemicals in women. Here we review our work and the work of others studying these mechanisms and propose a model for the ovarian action of di-(2-ethylhexyl) phthalate (DEHP). In vivo, DEHP (2 g/kg) causes decreased serum estradiol levels, prolonged estrous cycles, and no ovulations in adult, cycling rats. In vitro, monoethylhexyl phthalate (MEHP; the active metabolite of DEHP) decreases granulosa cell aromatase RNA message and protein levels in a dose-dependent manner. MEHP is unique among the phthalates in its suppression of aromatase and in its ability to activate peroxisome proliferator activated receptors (PPARs). We hypothesize that MEHP activates the PPARs to suppress aromatase in the granulosa cell. MEHP-, PPAR alpha-, and PPAR gamma specific ligands all similarly decreased estradiol production and RNA message levels of aromatase in vitro. Our model shows that MEHP acts on the granulosa cell by decreasing cAMP stimulated by follicle stimulating hormone and by activating the PPARs, which leads to decreased aromatase transcription. Thus, the environmental contaminant DEHP, through its metabolite MEHP, acts through a receptor-mediated signaling pathway to suppress estradiol production in the ovary, leading to anovulation. PMID- 12573896 TI - Effects of concentrated ambient particles on heart rate and blood pressure in pulmonary hypertensive rats. AB - Epidemiologic studies have shown that increased concentrations of ambient particles are associated with cardiovascular morbidity and mortality. However, the exact mechanisms remain unclear. Recent studies have revealed that particulate air pollution exposure is associated with indicators of autonomic function including heart rate, blood pressure, and heart rate variability. However, this association has not been clearly demonstrated in animal studies. To overcome the problems of wide variations in diseased animals and circadian cycles, we adopted a novel approach using a mixed-effects model to investigate whether ambient particle exposure was associated with changes in heart rate and blood pressure in pulmonary hypertensive rats. Male Sprague-Dawley rats were implanted with radiotelemetry devices and exposed to concentrated ambient particles generated by an air particle concentrator. The rats were held in nose only exposure chambers for 6 hr per day for 3 consecutive days and then rested for 4 days in each week during the experimental period of 5 weeks. These animals were exposed to concentrated particles during weeks 2, 3, and 4 and exposed to filtered air during weeks 1 and 5. The particle concentrations for tested animals ranged between 108 and 338 micro g/m(3). Statistical analysis using mixed-effects models revealed that entry and exit of exposure chamber and particle exposure were associated with changes in heart rate and mean blood pressure. Immediately after particle exposure, the hourly averaged heart rate decreased and reached the lowest at the first and second hour of exposure for a decrease of 14.9 (p < 0.01) and 11.7 (p = 0.01) beats per minute, respectively. The hourly mean blood pressure also decreased after the particle exposure, with a maximal decrease of 3.3 (p < 0.01) and 4.1 (p < 0.01) mm Hg at the first and second hour of exposure. Our results indicate that ambient particles might influence blood pressure and heart rate. PMID- 12573897 TI - Urinary levels of trichloroacetic acid, a disinfection by-product in chlorinated drinking water, in a human reference population. AB - Trichloroacetic acid (TCAA), a known mouse liver carcinogen and a possible human carcinogen, is found in chlorinated drinking water. We measured TCAA in archived urine samples from a reference population of 402 adults using isotope-dilution high-performance liquid chromatography-tandem mass spectrometry. TCAA was detected in 76% of the samples examined at concentrations ranging from < 0.5 micro g TCAA/L to more than 25 micro g/L; the 90th percentile concentration was 23 micro g/L (22 micro g TCAA/g creatinine); and the geometric mean and median concentrations were 2.9 micro g/L (2.6 micro g/g creatinine) and 3.3 micro g/L (3.2 micro g/g creatinine), respectively. The frequency of detection of TCAA in urban areas was higher than in rural areas (p = 0.00007), and sex and place of residence (i.e., urban vs. rural) were found to have a significant interaction in modulating the levels of TCAA (p = 0.012). Urban residents had higher mean levels of TCAA (men, 5.3 micro g/L, 3.8 micro g/g creatinine; women, 2.9 micro g/L, 2.8 micro g/g creatinine) than did rural residents (men, 2.2 micro g/L, 1.7 micro g/g creatinine; women, 2.6 micro g/L, 2.7 micro g/g creatinine). The higher frequency of detection of TCAA in urban than in rural areas and higher levels of TCAA among urban than among rural residents may reflect the fact that urban residents use primarily chlorinated water from public water supplies, whereas those in rural areas are more likely to obtain water from private wells, which typically are not chlorinated. PMID- 12573898 TI - Prevalence of non-insulin-dependent diabetes mellitus and related vascular diseases in southwestern arseniasis-endemic and nonendemic areas in Taiwan. AB - There is evidence indicating that ingestion of arsenic may predispose the development of diabetes mellitus in arsenic-endemic areas in Taiwan. However, the prevalence of diabetes and related vascular diseases in the entire southwestern arseniasis-endemic and nonendemic areas remains to be elucidated. We used the National Health Insurance Database for 1999-2000 to derive the prevalence of non insulin-dependent diabetes and related vascular diseases by age and sex among residents in southwestern arseniasis-endemic and nonendemic areas in Taiwan. The study included 66,667 residents living in endemic areas and 639,667 in nonendemic areas, all greater than or equal to 25 years of age. The status of diabetes and vascular diseases was ascertained through disease diagnosis and treatment prescription included in the reimbursement claims of clinics and hospitals. The prevalence of non-insulin-dependent diabetes, age- and gender-adjusted to the general population in Taiwan, was 7.5% (95% confidence interval, 7.4-7.7%) in the arseniasis-endemic areas and 3.5% (3.5-3.6%) in the nonendemic areas. Among both diabetics and nondiabetics, higher prevalence of microvascular and macrovascular diseases was observed in arseniasis-endemic than in the nonendemic areas. Age- and gender-adjusted prevalence of microvascular disease in endemic and nonendemic areas was 20.0% and 6.0%, respectively, for diabetics, and 8.6% and 1.0%, respectively, for nondiabetics. The corresponding prevalence of macrovascular disease was 25.3% and 13.7% for diabetics, and 12.3% and 5.5% for nondiabetics. Arsenic has been suggested to increase the risk of non-insulin-dependent diabetes mellitus and its related micro- and macrovascular diseases. PMID- 12573899 TI - Personal PM2.5 exposure and markers of oxidative stress in blood. AB - Ambient particulate air pollution assessed as outdoor concentrations of particulate matter less than or equal to 2.5 micro m in diameter (PM(2.5)) in urban background has been associated with cardiovascular diseases at the population level. However, the significance of individual exposure and the involved mechanisms remain uncertain. We measured personal PM(2.5) and carbon black exposure in 50 students four times in 1 year and analyzed blood samples for markers of protein and lipid oxidation, for red blood cell (RBC) and platelet counts, and for concentrations of hemoglobin and fibrinogen. We analyzed protein oxidation in terms of gamma-glutamyl semialdehyde in hemoglobin (HBGGS) and 2 aminoadipic semialdehyde in hemoglobin (HBAAS) and plasma proteins (PLAAS), and lipid peroxidation was measured as malondialdehyde (MDA) in plasma. Median exposures were 16.1 micro g/m(3) for personal PM(2.5) exposure, 9.2 micro g/m(3) for background PM(2.5) concentration, and 8.1 X 10(-6)/m for personal carbon black exposure. Personal carbon black exposure and PLAAS concentration were positively associated (p < 0.01), whereas an association between personal PM(2.5) exposure and PLAAS was only of borderline significance (p = 0.061). A 3.7% increase in MDA concentrations per 10 micro g/m(3) increase in personal PM(2.5) exposure was found for women (p < 0.05), whereas there was no significant relationship for the men. Similarly, positive associations between personal PM(2.5)exposure and both RBC and hemoglobin concentrations were found only in women (p < 0.01). There were no significant relationships between background PM(2.5) concentration and any of the biomarkers. This suggests that exposure to particles in moderate concentrations can induce oxidative stress and increase RBCs in peripheral blood. Personal exposure appears more closely related to these biomarkers potentially related to cardiovascular disease than is ambient PM(2.5) background concentrations. PMID- 12573900 TI - Perchloroethylene-contaminated drinking water and the risk of breast cancer: additional results from Cape Cod, Massachusetts, USA. AB - In 1998 we published the results of a study suggesting an association between breast cancer and perchloroethylene (PCE; also called tetrachloroethylene) exposure from public drinking water. The present case-control study was undertaken to evaluate this association further. The cases were composed of female residents of eight towns in the Cape Cod region of Massachusetts who had been diagnosed with breast cancer from 1987 through 1993 (n = 672). Controls were composed of demographically similar women from the same towns (n = 616). Women were exposed to PCE when it leached from the vinyl lining of water distribution pipes from the late 1960s through the early 1980s. A relative delivered dose of PCE that entered a home was estimated using an algorithm that took into account residential history, water flow, and pipe characteristics. Small to moderate elevations in risk were seen among women whose exposure levels were above the 75th and 90th percentiles when 0-15 years of latency were considered (adjusted odds ratios, 1.5-1.9 for > 75th percentile, 1.3-2.8 for > 90th percentile). When data from the present and prior studies were combined, small to moderate increases in risk were also seen among women whose exposure levels were above the 75th and 90th percentiles when 0-15 years of latency were considered (adjusted odds ratios, 1.6-1.9 for > 75th percentile, 1.3-1.9 for > 90th percentile). The results of the present study confirm those of the previous one and suggest that women with the highest PCE exposure levels have a small to moderate increased risk of breast cancer. PMID- 12573901 TI - Aggressive behavior and serum testosterone concentration during the maturation process of male mice: the effects of fetal exposure to bisphenol A. AB - The relationship between exposure to endocrine-disrupting chemicals (EDs) and risk to reproductive organs is well documented, but the influence of EDs on behavioral development has not been studied. In this study we evaluated the effect of fetal exposure to bisphenol A, which mimics estrogenic activity, on aggressive behavior and hormonal change in male mice. On gestation days 11-17, female mice were fed bisphenol A at 2 ng/g or 20 ng/g of body weight (environmentally relevant concentration). Aggression rating and blood sampling of the offspring were done at 8, 12, and 16 weeks of age. Aggression scores increased significantly (p < 0.01) at 8 weeks of age in male mice exposed to bisphenol A at both the 2 ng/g and 20 ng/g concentrations compared with a control group, but no difference was found after 12 weeks. Relative testis weight (per gram of body weight) was significantly lower at 8 and 12 weeks in mice treated with 2 ng/g than in controls (p < 0.05) and was significantly lower at 12 weeks in mice treated with 20 ng/g than in controls (p < 0.01). The serum testosterone concentration in treated mice was not significantly different from that in controls. These results demonstrate that bisphenol A temporarily activated aggressive behavior in mice at 8 weeks of age and that low doses of bisphenol A interfered with the normal development of reproductive organs. The mechanism activating this aggressive behavior was not elevated testosterone concentration. PMID- 12573902 TI - Risk of non-Hodgkin's lymphoma and prediagnostic serum organochlorines: beta hexachlorocyclohexane, chlordane/heptachlor-related compounds, dieldrin, and hexachlorobenzene. AB - Increases in non-Hodgkin's lymphoma (NHL) incidence and mortality rates during the past few decades remain largely unexplained. Studies suggest that organochlorine pesticides may contribute to an increased risk of NHL. In 1974, serum samples were obtained from 25,802 participants in the Campaign Against Cancer and Stroke in Washington County, Maryland (USA), and cryopreserved for future study. We measured prediagnostic levels of chlordane, lindane (gamma hexachlorocyclohexane), beta-hexachlorocyclohexane, transnonachlor, heptachlor, heptachlor epoxide, oxychlordane, dieldrin, and hexachlorobenzene in serum samples of 74 cases of NHL and 147 matched controls. Previously, we found an association between NHL and serum levels of total PCBs (polychlorinated biphenyls), but not DDT (dichlorodiphenyltrichloroethane) and related compounds. In this instance, there was no evidence of an association between NHL risk and serum levels of any of the individual lipid- and recovery-corrected organochlorines that we evaluated, nor of the summed chlordane-related compounds (transnonachlor, heptachlor, heptachlor epoxide, oxychlordane). These findings do not support the hypothesis that the organochlorine compounds included in this study are strongly linked to the development of NHL. The possibility of a weak association cannot be excluded by these data. PMID- 12573903 TI - The high cost of improper removal of lead-based paint from housing: a case report. AB - The costs of lead-based paint hazard control in housing are well documented, but the costs of cleanup after improper, inherently dangerous, methods of removing lead-based paint are not. In this article we report a case of childhood lead poisoning and document the costs of decontamination after uncontained power sanding was used to remove paint down to bare wood from approximately 3,000 ft(2) of exterior siding on a large, well-maintained 75-year-old house in a middle income neighborhood. After the uncontrolled removal of lead-based paint, interior dust lead levels ranged from 390 to 27,600 micro g Pb/ft(2) (on floors and windowsills) and bare soil lead levels ranged from 360 ppm in the yard to 3,900 ppm along the foundation to 130,000 ppm in the child's play area, well above applicable U.S. Department of Housing and Urban Development/U.S. Environmental Protection Agency standards. The hard costs of decontamination were over $195,000, which greatly exceeds the incremental cost of incorporating lead-safe work practices into repainting. This case report highlights the need to incorporate lead-safe work practices into routine repainting, remodeling, and other renovation and maintenance jobs that may disturb lead-based paint. PMID- 12573904 TI - Hair and toenail arsenic concentrations of residents living in areas with high environmental arsenic concentrations. AB - Surface soil and groundwater in Australia have been found to contain high concentrations of arsenic. The relative importance of long-term human exposure to these sources has not been established. Several studies have investigated long term exposure to environmental arsenic concentrations using hair and toenails as the measure of exposure. Few have compared the difference in these measures of environmental sources of exposure. In this study we aimed to investigate risk factors for elevated hair and toenail arsenic concentrations in populations exposed to a range of environmental arsenic concentrations in both drinking water and soil as well as in a control population with low arsenic concentrations in both drinking water and soil. In this study, we recruited 153 participants from areas with elevated arsenic concentrations in drinking water and residential soil, as well as a control population with no anticipated arsenic exposures. The median drinking water arsenic concentrations in the exposed population were 43.8 micro g/L (range, 16.0-73 micro g/L) and median soil arsenic concentrations were 92.0 mg/kg (range, 9.1-9,900 mg/kg). In the control group, the median drinking water arsenic concentration was below the limit of detection, and the median soil arsenic concentration was 3.3 mg/kg. Participants were categorized based on household drinking water and residential soil arsenic concentrations. The geometric mean hair arsenic concentrations were 5.52 mg/kg for the drinking water exposure group and 3.31 mg/kg for the soil exposure group. The geometric mean toenail arsenic concentrations were 21.7 mg/kg for the drinking water exposure group and 32.1 mg/kg for the high-soil exposure group. Toenail arsenic concentrations were more strongly correlated with both drinking water and soil arsenic concentrations; however, there is a strong likelihood of significant external contamination. Measures of residential exposure were better predictors of hair and toenail arsenic concentrations than were local environmental concentrations. PMID- 12573905 TI - Maternal blood lead concentration, diet during pregnancy, and anthropometry predict neonatal blood lead in a socioeconomically disadvantaged population. AB - To determine the influences of maternal diet and nutrition during pregnancy on the blood lead level of neonates, we conducted a study of mother-infant pairs from lower socioeconomic circumstances living in Albany County, New York. Maternal blood lead (MBPb), anthropometry, and diet were assessed in each trimester. Neonates' blood lead (NBPb) levels were low (geometric mean = 1.58 micro g/dL), and none had elevated blood lead. More than 50% of the mothers had intakes below the recommended dietary allowances for zinc, calcium, iron, vitamin D, and kilocalories. As expected, MBPb was strongly and positively related to NBPb. Among the anthropometric measures of maternal nutritional status, variables measuring gain in weight and arm circumference were negatively related to NBPb. In multivariable models reflecting different analytic strageties and including MBPb, anthropometry, and sociodemographic characteristics, dietary intakes of iron and vitamin D were negatively related to NBPb. The effect of zinc varied substantially depending on model covariates. Effects of dietary constituents are difficult to distinguish, given the intercorrelated nature of nutrients in the diet. Nevertheless, the influences of maternal anthropometric variables, iron, and vitamin D on neonatal lead levels are clear in our analyses. PMID- 12573906 TI - Effects of transplacental exposure to environmental pollutants on birth outcomes in a multiethnic population. AB - Inner-city, minority populations are high-risk groups for adverse birth outcomes and also are more likely to be exposed to environmental contaminants, including environmental tobacco smoke (ETS), polycyclic aromatic hydrocarbons (PAHs), and pesticides. In a sample of 263 nonsmoking African-American and Dominican women, we evaluated the effects on birth outcomes of prenatal exposure to airborne PAHs monitored during pregnancy by personal air sampling, along with ETS estimated by plasma cotinine, and an organophosphate pesticide (OP) estimated by plasma chlorpyrifos (CPF). Plasma CPF was used as a covariate because it was the most often detected in plasma and was highly correlated with other pesticides frequently detected in plasma. Among African Americans, high prenatal exposure to PAHs was associated with lower birth weight (p = 0.003) and smaller head circumference (p = 0.01) after adjusting for potential confounders. CPF was associated with decreased birth weight and birth length overall (p = 0.01 and p = 0.003, respectively) and with lower birth weight among African Americans (p = 0.04) and reduced birth length in Dominicans (p < 0.001), and was therefore included as a covariate in the model with PAH. After controlling for CPF, relationships between PAHs and birth outcomes were essentially unchanged. In this analysis, PAHs and CPF appear to be significant independent determinants of birth outcomes. Further analyses of pesticides will be carried out. Possible explanations of the failure to find a significant effect of PAHs in the Hispanic subsample are discussed. This study provides evidence that environmental pollutants at levels currently encountered in New York City adversely affect fetal development. PMID- 12573907 TI - Residential proximity to traffic and adverse birth outcomes in Los Angeles county, California, 1994-1996. AB - We reported previously that increases in ambient air pollution in the Los Angeles basin increased the risk of low weight and premature birth. However, ambient concentrations measured at monitoring stations may not take into account differential exposure to pollutants found in elevated concentrations near heavy traffic roadways. Therefore, we used an epidemiologic case-control study design to examine whether residential proximity to heavy-traffic roadways influenced the occurrence of low birth weight (LBW) and/or preterm birth in Los Angeles County between 1994 and 1996. We mapped subject home locations at birth and estimated exposure to traffic-related air pollution using a distance-weighted traffic density (DWTD) measure. This measure takes into account residential proximity to and level of traffic on roadways surrounding homes. We calculated odds ratios (ORs) and risk ratios (RRs) for being LBW and/or preterm per quintile of DWTD. The clearest exposure-response pattern was observed for preterm birth, with an RR of 1.08 [95% confidence interval (CI), 1.01-1.15] for infants in the highest DWTD quintile. Although higher risks were observed for LBW infants, exposure-response relations were less consistent. Examining the influence of season, we found elevated risks primarily for women whose third trimester fell during fall/winter months (OR(term LBW) = 1.39; 95% CI, 1.16-1.67; OR(preterm and LBW) = 1.24; 95% CI = 1.03-1.48; RR(all preterm) = 1.15; 95% CI, 1.05-1.26), and exposure-response relations were stronger for all outcomes. This result is consistent with elevated pollution in proximity to sources during more stagnant air conditions present in winter months. Our previous research and these latest results suggest exposure to traffic-related pollutants may be important. PMID- 12573908 TI - Modification of immune function through exposure to dietary aflatoxin in Gambian children. AB - Aflatoxins are immunotoxins that frequently contaminate staple foods in The Gambia and other parts of sub-Saharan Africa, resulting in high exposure throughout life. Impaired infant immune system development may be a key predictor of mortality from infectious disease. In this study we aimed to determine the effect of dietary aflatoxin exposure on a number of immune parameters in Gambian children. A cohort of 472 Gambian children 6-9 years of age was recruited. Serum aflatoxin-albumin (AF-alb) adducts were analyzed to provide a measure of exposure. Immune parameters included secretory IgA (sIgA) in saliva, cell mediated immunity (CMI), determined using the CMI multitest where test antigens are applied to the skin, and antibody responses to both rabies and pneumococcal polysaccharide vaccines. Birth weight, current anthropometry, and micronutrient status were also recorded. AF-alb adducts were detected in 93% of the children (geometric mean level 22.3 pg/mg; range 5-456 pg/mg). AF-alb level was strongly influenced by month of sampling. In a multivariable analysis, sIgA was markedly lower in children with detectable AF-alb compared with those with nondetectable levels [50.4 micro g/mg protein (95% confidence interval [CI] 48.0-52.8) and 70.2 micro g/mg protein (95% CI 61.1-79.2), respectively; p < 0.0001]. Antibody response to one of four pneumococcal serotypes, but not rabies vaccine, was weakly associated with higher levels of AF-alb. There was no association between CMI responses to test antigens and AF-alb. These data confirm that children in rural Gambia are frequently exposed to high levels of aflatoxin. The study provides evidence that sIgA in saliva may be reduced because of dietary levels of aflatoxin exposure. Given the high burden of infection-related mortality in West Africa, further investigation of the immune effects of aflatoxin exposure in children is merited. PMID- 12573909 TI - Animal models to detect allergenicity to foods and genetically modified products: workshop summary. AB - Respiratory allergy and allergy to foods continue to be important health issues. There is evidence to indicate that the incidence of food allergy around the world is on the rise. Current estimates indicate that approximately 5% of young children and 1-2% of adults suffer from true food allergy (Kagan 2003). Although a large number of in vivo and in vitro tests exist for the clinical diagnosis of allergy in humans, we lack validated animal models of allergenicity. This deficiency creates serious problems for regulatory agencies and industries that must define the potential allergenicity of foods before marketing. The emergence of several biotechnologically derived foods and industrial proteins, as well as their potential to sensitize genetically predisposed populations to develop allergy, has prompted health officials and regulatory agencies around the world to seek approaches and methodologies to screen novel proteins for allergenicity. PMID- 12573910 TI - Food allergy: an overview. AB - Food allergy affects between 5% and 7.5% of children and between 1% and 2% of adults. The greater prevalence of food allergy in children reflects both the increased predisposition of children to develop food allergies and the development of immunologic tolerance to certain foods over time. Immunoglobulin (Ig) E-mediated food allergies can be classified as those that persist indefinitely and those that are predominantly transient. Although there is overlap between the two groups, certain foods are more likely than others to be tolerated in late childhood and adulthood. The diagnosis of food allergy rests with the detection of food-specific IgE in the context of a convincing history of type I hypersensitivity-mediated symptoms after ingestion of the suspected food or by eliciting IgE-mediated symptoms after controlled administration of the suspected food. Presently, the only available treatment of food allergies is dietary vigilance and administration of self-injectable epinephrine. PMID- 12573912 TI - Assessment of the allergic potential of food protein extracts and proteins on oral application using the brown Norway rat model. AB - The need for widely accepted and validated animal models to test the potential allergenicity and potency of novel (biotechnology-derived) proteins has become an important issue for their safety evaluation. In this article, we summarize the results of the development of an oral sensitization protocol for food proteins in the rat. Young Brown Norway rats were exposed to either various purified allergenic proteins (e.g., ovalbumin, partly purified), a whole food (cow's milk), or total protein extracts (hen's egg white, peanut) by daily gavage dosing during 42 days without the use of an adjuvant. The results showed that Brown Norway rats can be sensitized orally to the various allergenic food proteins tested, resulting in antigen-specific immunoglobulin (Ig) G and IgE responses, without the use of adjuvants. Animals orally exposed to cow's milk or total protein extracts of egg white also developed specific IgE and IgG antibodies that recognized the same proteins compared with antibodies from patients allergic to egg white or cow's milk. We also studied local and systemic immune-mediated effects. In ovalbumin-sensitized rats, some clinical symptoms of food allergy were studied upon an oral challenge with ovalbumin. The results demonstrated that gut permeability was increased and that in some animals breathing frequency and systolic blood pressure were temporarily decreased. The results obtained show that the Brown Norway rat provides a suitable animal model for food allergy research and for the study of relative allergenicity of existing and novel food proteins. PMID- 12573911 TI - Assessment of the inherent allergenic potential of proteins in mice. AB - There is considerable interest in the design of approaches that will permit the accurate identification and characterization of proteins that have the inherent potential to induce sensitization and cause food allergy. Among the methods used currently as part of such assessments are consideration of structural similarity to, or amino acid sequence homology with, known human allergens; whether there exists immunologic cross-reactivity with known allergens; and measurement of resistance to proteolytic digestion in a simulated gastric fluid. Although such approaches provide information that will contribute to a safety assessment, they do not--either individually or collectively--provide a direct evaluation of the ability of a novel protein to cause allergic sensitization. For this reason, work is in progress to design and evaluate suitable animal models that will provide a more holistic assessment of allergenic potential. In this laboratory, the approach we have taken has been to examine the characteristics of immune responses induced in mice following parenteral (intraperitoneal) exposure to test proteins. The basis of this method is to determine simultaneously the overall immunogenic potential of proteins [measured as a function of immunoglobulin (Ig) G antibody responses] and to compare this with their ability to provoke IgE antibody production, IgE being the antibody that effects allergic sensitization. Although this approach has not yet been evaluated fully, the results available to date suggest that it will be possible to distinguish proteins that have the inherent potential to induce allergic sensitization from those that do not. In this article we summarize progress to date in the context of the scientific background against which such methods are being developed. PMID- 12573913 TI - Nonmurine animal models of food allergy. AB - Food allergy can present as immediate hypersensitivity [manifestations mediated by immunoglobulin (Ig)E], delayed-type hypersensitivity (reactions associated with specific T lymphocytes), and inflammatory reactions caused by immune complexes. For reasons of ethics and efficacy, investigations in humans to determine sensitization and allergic responses of IgE production to innocuous food proteins are not feasible. Therefore, animal models are used a) to bypass the innate tendency to develop tolerance to food proteins and induce specific IgE antibody of sufficient avidity/affinity to cause sensitization and upon reexposure to induce an allergic response, b) to predict allergenicity of novel proteins using characteristics of known food allergens, and c) to treat food allergy by using immunotherapeutic strategies to alleviate life-threatening reactions. The predominant hypothesis for IgE-mediated food allergy is that there is an adverse reaction to exogenous food proteins or food protein fragments, which escape lumen hydrolysis, and in a polarized helper T cell subset 2 (Th2) environment, immunoglobulin class switching to allergen-specific IgE is generated in the immune system of the gastrointestinal-associated lymphoid tissues. Traditionally, the immunologic characterization and toxicologic studies of small laboratory animals have provided the basis for development of animal models of food allergy; however, the natural allergic response in large animals, which closely mimic allergic diseases in humans, can also be useful as models for investigations involving food allergy. PMID- 12573915 TI - A human dendritic cell-based method to identify CD4+ T-cell epitopes in potential protein allergens. AB - We developed an assay to determine the location of immunodominant CD4(+) T-cell epitopes in any protein. The method uses CD4(+) T cells from community donors in conjunction with dendritic cells derived in vitro. Synthetic peptides constructed to describe the sequence of the protein of interest are cocultured with dendritic cells and CD4(+) T cells, and T-cell proliferation is measured. Data are compiled over a large replicate of human donors to pinpoint immunodominant, usually promiscuous epitope regions. We have applied this technique to a known food allergen, the Brazil nut 2S storage globulin protein, and to two potential food allergens, the Cry1Ab and Cry3Aa proteins. We show epitope data for these three proteins. This assay can be used as a tool to guide the selection and qualification of future potential food transgenes. PMID- 12573914 TI - Identification of antigenic epitopes on human allergens: studies with HLA transgenic mice. AB - Environmental factors play an important role in the rise and manifestation of allergic conditions in genetically predisposed subjects. Increased exposure to indoor/outdoor allergens is a significant factor in the development of allergic sensitization and asthma. Recently, strong relationships between the immune response to several highly purified allergens and specific human leukocyte antigen (HLA)-DQ and -DR haplotypes have been reported. The major antigens from clinically important allergens have been cloned and sequenced. However, whether innate structural features of major allergens or peculiar immune recognition of these molecules contribute to the overly robust immune responses is not known. We generated and used transgenic (tg) mice expressing single HLA class II transgene(s) to characterize the allergen epitopes presented by particular HLA class II molecules. Next, we generated in vivo models for asthma in the HLA tg mice by intranasal challenge with allergenic extracts. Furthermore, we used a single epitope to induce an allergic lung inflammation. Our system offers a sophisticated technique for systematically identifying the genetic (individual human class II) and antigenic (individual allergenic epitopes) basis of asthma sensitivity and has important implications for new treatment strategies. PMID- 12573916 TI - Reversing urban decay: brownfield redevelopment and environmental health. AB - While the United States government concentrates more of its political and financial resources on fighting terrorism, the continuing decay of older cities and industrial suburbs has fallen far down on the national political priority agenda. An exception is the redevelopment of so-called brownfields, which are abandoned, idled, or underutilized factories, railroad yards, bus stations, garages, electricity-generating stations, and other commercial facilities. A modest national government program to identify, clean up, and redevelop brownfields into job fields began during the administration of Bill Clinton and has continued into the George W. Bush administration (Powers et al. 2000; Simons 1998; Van Horn et al. 1999). The political reasons are apparent: Developing brownfields is a politically acceptable method of stimulating private enterprise, local government, and community groups into building new businesses, housing, and community facilities. Also, brownfields projects have a beginning and an end; the national government does not have an indefinite responsibility. In contrast, social assistance programs that grew during the 1960s and proliferated for more than three decades have been politically portrayed by some as give-away programs that build dependency with no ending. Whether this characterization of social programs is morally or empirically justified, the reality is that in today's political environment brownfields redevelopment is a politically acceptable way of helping distressed urban areas. PMID- 12573917 TI - Chronic lyme disease: psychogenic fantasy or somatic infection? AB - Sigal and Hassett published an article about Lyme disease in the EHP Supplements (Sigal and Hassett 2002), suggesting that chronic Lyme disease is "psychogenic." I do not think that Sigal and Hassett, non-psychiatrists, are qualified to speak about psychiatric matters. I, however, actually have had the disease, which they characterize as "medically unexplained," for over 25 years and have 15 years of experience as a patient advocate and educator. I beg to differ. PMID- 12573918 TI - Chronic Lyme disease: it's not all in our heads. AB - Those of us with chronic Lyme disease are not at all confused, as suggested by Sigal and Hassett (2002). We know from years of experience that we have real, specific symptoms that are usually painful and disabling and include severe headaches, crippling arthritis, and heart palpitations, which lead to serious heart disease. Many of us know that our symptoms are kept in check while we are on antibiotics, but they painfully reappear when the antibiotics are withdrawn. Just because the medical community cannot detect a specific causative bacterium and managed health care companies want to maximize profits doesn't mean that those of us afflicted with this terrible condition are delusional and not truly benefiting from antibiotic treatment. We are not all crazy; we are sick and we should not be required to prove it to get medical care. PMID- 12573920 TI - Cancer in beluga from the St. Lawrence estuary. AB - Martineau et al. (2002) reported that St. Lawrence beluga (SLB) have high cancer rates. Unfortunately, errors in their interpretation of the data have led them to overstate the importance of cancer and its links to environmental sources. PMID- 12573922 TI - Double jeopardy? PMID- 12573923 TI - A luxury tax on energy. PMID- 12573924 TI - Give me shelter: the global housing crisis. AB - In both developed and developing countries around the world, the health of significant numbers of people is adversely affected by a lack of adequate housing. Large-scale migrations into already crowded developing nation cities compound existing health problems associated with poor indoor air quality, contaminated drinking water, and limited sanitation infrastructure. In the developed world, lead exposure, indoor air quality, and asthma are among the most serious and costly housing-related health risks. PMID- 12573925 TI - Mold insurance: crafting coverage for a spreading problem. AB - Mold contamination is a growing concern for homeowners in terms of both physical health and insurance. Health experts, although they concede that exposure to mold can cause respiratory illnesses, are calling for further research into other mold related health effects and for development of standards for mold sampling and data analysis. The insurance industry is grappling with how---and whether---to provide coverage for household damage caused by mold, while some state and federal legislators are working to pass laws regarding mold testing and insurance. PMID- 12573926 TI - Solar flair. AB - Design innovations and government-sponsored financial incentives are making solar energy increasingly attractive to homeowners and institutional customers such as school districts. In particular, the passive solar design concept of daylighting is gaining favor among educators due to evidence of improved performance by students working in daylit classrooms. Electricity-generating photovoltaic systems are also becoming more popular, especially in states such as California that have high electric rates and frequent power shortages. To help spread the word about solar power, the U.S. Department of Energy staged its first-ever Solar Decathlon in October 2002. This event featured solar-savvy homes designed by 14 college teams. PMID- 12573927 TI - The information-processing paradigm: a valuable framework for clinical child and adolescent psychology. AB - Provides an introduction to the special section on information-processing (I-P) factors in child and adolescent psychopathology. First, we describe the I-P paradigm and summarize its central tenets, presenting examples of past research that illustrate the heuristic value of the paradigm. Next, we discuss the potential benefits of the I-P paradigm for the field of clinical child and adolescent psychology. Finally, we present an overview of the articles in the special section. PMID- 12573928 TI - Patterns of processing bias for emotional information across clinical disorders: a comparison of attention, memory, and prospective cognition in children and adolescents with depression, generalized anxiety, and posttraumatic stress disorder. AB - This study investigated theoretical claims that different emotional disorders are associated with different patterns of cognitive bias, both in terms of the cognitive processes involved and the stimulus content that is preferentially processed. These claims were tested by comparing clinically anxious (generalized anxiety disorder [GAD], posttraumatic stress disorder [PTSD]) and clinically depressed children and adolescents on a range of cognitive tasks measuring attention, memory, and prospective cognition, with both threat-related and depressogenic stimulus materials. The results did reveal some relative specificity of processing in that the anxious participants exhibited a greater selective attentional bias for threat relative to depressogenic material with no such difference being apparent in the depressed sample. However, this bias was only clear-cut on a dot-probe measure of attentional processing and not on a modified Stroop measure, and indeed threat-related bias on the 2 tasks was uncorrelated. On the prospective cognition task, anxious participants exhibited an other-referent bias in their risk estimations regarding future negative events that was absent in the depressed sample. No specificity effects were evident on the memory task. The results are discussed in terms of the strengths and weaknesses of carrying out direct comparisons across groups and tasks versus drawing conclusions from overall patterns across multiple studies. PMID- 12573929 TI - Autobiographical memory specificity and trauma in inpatient adolescents. AB - Investigated the relation between memory specificity and self-reported trauma, depressive symptoms, and other emotional characteristics in a group of adolescent inpatients. Research with adults has shown that clinical depression is associated with a difficulty in retrieving specific autobiographical memories in response to cue words, and this pattern of overgeneral memory is related to the prognosis of depression. Research has also shown a clear positive association between self reported trauma and overgeneral memory. This study's results showed that higher levels of trauma--both in terms of total number and in terms of severity or related distress--were associated with reduced autobiographical memory specificity (AM). None of the other emotional variables, such as depression, anxiety, worry, hopelessness, or subjective stress, were significantly related to the retrieval of specific memories. The results are reconcilable with Williams' (1996) developmental idea that a pattern of overgeneral AM retrieval originates in childhood trauma as a way of regulating affect. PMID- 12573930 TI - Cognitive interference due to food cues in childhood obesity. AB - Investigated specific information-processing biases for food-relevant stimuli in 34 children, Mage = 13 years, SD = 2 years, M = 177% of ideal weight, SD = 24%, who were completing a residential cognitive-behavioral treatment program for severe obesity and 40 matched control children who were not obese, Mage = 13 years, SD = 2 years, M = 103% of ideal weight, SD = 13%. Participants completed a computerized modified Stroop task, which included food words, negative-emotion words, and control words. Results indicated that children in the obesity group displayed a specific interference effect for food words. It is hypothesized that the observed bias in information processing reflects hypersensitivity for food cues, which can initiate or maintain dysfunctional eating behavior. PMID- 12573931 TI - Suppressed attention to rejection, ridicule, and failure cues: a unique correlate of reactive but not proactive aggression in youth. AB - Tested the hypothesis that reactive aggression (RA) but not proactive aggression (PA) should be associated with heightened attention to rejection, ridicule, and failure cues. In addition to a reaction time measure of selective attention, participants also completed a vignette-based interview regarding their interpretation of ambiguous social situations, and children, parents, and teachers completed questionnaire measures of child aggression and related variables. Consistent with predictions, RA but not PA was related to biased attention for rejection, ridicule, and failure cues. However, contrary to expectation, heightened RA scores were associated with suppressed rather than enhanced attention to such cues. Despite the unexpected direction of this attentional bias, as predicted it was significantly related to the well established tendency of aggressive children to interpret ambiguous social situations as threatening, which was also uniquely related to RA. Further, the correlation between suppressed attention and RA was fully mediated by interpretation bias. PMID- 12573932 TI - Negative Feelings Exacerbate Hostile Attributions of Intent in Highly Aggressive Boys. AB - Tested the hypothesis that aggressive boys' tendency to attribute hostile intentions to peers is exacerbated in a negative emotional state. Twenty-nine highly aggressive boys in special education, 12 moderately aggressive boys in regular education, and 16 nonaggressive boys in regular education inferred peers' intentions in 8 vignettes concerning ambiguous provocation by peers. Mild negative emotions were induced by unjust loss of a manipulated computer game. Half the vignettes were completed in this negative emotional state. After completion of all vignettes, the game was played again and won to reinduce positive feelings. Self-ratings of feelings obtained throughout the study showed the manipulations consecutively induced negative and positive feelings. Negative feelings increased hostile attribution of intent in the highly aggressive group. Highly and moderately aggressive boys responded more aggressively than nonaggressive boys. PMID- 12573933 TI - Callous-unemotional traits, impulsivity, and emotional processing in adolescents with antisocial behavior problems. AB - Examined the emotional reactivity of adolescents with antisocial behavior problems using a lexical decision paradigm. Evidence from adult forensic samples indicates that psychopathic traits are associated with abnormalities in the processing of emotional stimuli. In an attempt to extend these findings earlier in development, this association was tested in a sample of adolescents (mean age = 16.01; SD = 1.32) referred to a diversion program for delinquent behavior. Emotional processing was assessed by comparing recognition time for emotional words, both positive and negative, to recognition time for nonemotional words. Consistent with adult findings, the callous-unemotional (CU) dimension of psychopathy was associated with slower reaction times to negative words. In contrast, problems of impulse control were associated with faster recognition times for negative emotional words. These findings suggest that different patterns of emotional reactivity may characterize distinct subgroups of youth with antisocial behavior problems. PMID- 12573934 TI - Research on information-processing factors in child and adolescent psychopathology: a critical commentary. AB - Provides a critical commentary on the state-of-the-art of research on information processing (I-P) factors in clinical child and adolescent psychology. The articles in this special section amply demonstrate the value of the I-P paradigm as a heuristic framework for conceptualizing and studying the role(s) of cognitive factors in the etiology and maintenance of child and adolescent psychopathology. However, the current status of such research also reflects a number of limitations that warrant consideration if the potential value of the I P paradigm is to be fully realized. Specifically, understanding the role(s) played by such factors is impeded by a variety of insufficiently addressed methodological and psychometric issues, as well as by insufficiently articulated theories regarding such factors. These issues are particularly challenging for child and adolescent psychopathology researchers because of the complexities added by development. The value of I-P theories of childhood and adolescent psychopathology will be considerably enhanced if these issues are more fully considered in future research. PMID- 12573935 TI - Family instability and young adolescent maladjustment: the mediating effects of parenting quality and adolescent appraisals of family security. AB - Examined relations among family instability and adolescent's psychological functioning using family models of children's emotional security in a sample of 220 young adolescents and their primary caregivers. Primary caregiver reports of family instability were associated with multiple informant measures of adolescent internalizing and externalizing symptoms. Findings from structural equation models supported the hypothesis that family instability increases adolescent risk for psychological problems by directly undermining their insecure appraisals of the family. Results also supported a pathway whereby family instability predicted parenting difficulties and parenting difficulties, in turn, indirectly predicted adolescents' internalizing and externalizing symptoms through its association with lower levels of perceived insecurity in the family. Results are discussed in relation to how they advance process-oriented conceptualizations of family instability. PMID- 12573936 TI - Social self-efficacy and behavior problems in maltreated and nonmaltreated children. AB - Investigated the relations among child maltreatment, children's social self efficacy, and behavioral adjustment. Data were collected on 305 maltreated and 195 non-maltreated children from low-income families (ages 5 to 12 years) who were assessed on perceived social self-efficacy and evaluated by camp counselors on internalizing and externalizing symptomatology. Younger (< 8 years) maltreated children exhibited inflated levels of perceived self-efficacy in confictual peer interactions compared to younger nonmaltreated children. Younger maltreated children with higher levels of social self-efficacy showed significantly less internalizing behaviors compared to younger maltreated children with lower levels of social self-efficacy. For older children (> 8 years), regardless of maltreatment status, higher levels of perceived social self-efficacy in conflict situations were related to lower levels of internalizing symptomatology. The results are discussed as suggestive of the role of children's social self efficacy as a protective factor in the link between maltreatment and internalizing symptomatology. PMID- 12573937 TI - The impact of late-afternoon stimulant dosing for children with ADHD on parent and parent-child domains. AB - Examined the impact of late-afternoon stimulant dosing on parent and parent-child domains. Twenty-one children with attention deficit hyperactivity disorder (ADHD) participated in a double-blind, placebo-controlled medication assessment comparing varying doses of methylphenidate (MPH) and Adderall in the context of an intensive treatment program. Children received varying doses of stimulant medication, including active medication or placebo at 3:30 p.m. daily. Parent ratings of their mood, pleasantness of parent-child interactions, and perceived parenting effectiveness and successfulness were obtained each evening. Effective medication doses had a beneficial impact on parent-child domains (e.g., parent child interactions) but did not significantly affect non-child-related parent variables (e.g., negative affect [NA]). Findings suggested that doses with beneficial effects lasting into the evening improved parent reports of the pleasantness of parent-child interactions but were insufficient to produce positive changes in parent functioning. PMID- 12573938 TI - Emotional security with teachers and children's stress reactivity: a comparison of special-education and regular-education classrooms. AB - Examined children's exposure and reactivity to negative peer and teacher events in special-education and regular-education classrooms. Participants were 40 children in regular classrooms and 20 children classified as seriously emotionally disturbed (SED) in special-education classrooms. Children completed 7 days of diary data over the course of the school year. SED children reported higher rates of exposure to negative teacher and peer events than comparison children. The self-esteem of both SED and comparison children was reactive to negative peer events in the classroom, but emotional security with teacher (EST) reduced this reactivity. The self-esteem of SED children was also reactive to negative teacher events but, EST, once again, reduced reactivity to negative events. Finally, SED children's perception of maternal warmth moderated their self-esteem reactivity and was concordant with their views of teachers. Results are discussed in light of treatment implications for SED children. PMID- 12573939 TI - The relation of narcissism and self-esteem to conduct problems in children: a preliminary investigation. AB - Investigated several possible models to explain the seemingly discrepant relations between self-esteem and conduct problems, as both low self-esteem and exaggerated levels of self-esteem, thought to be captured by narcissism, have been associated with aggressive and antisocial behavior. Our sample consisted of 98 nonreferred children (mean age = 11.9 years; SD = 1.68 years) recruited from public schools to oversample children at risk for severe aggressive and antisocial behavior. Results indicated that certain aspects of narcissism (i.e., those indicating a need to be evaluated well by, and obtain status over, others) were particularly predictive of maladaptive characteristics and outcomes such as low self-esteem, callous-unemotional (CU) traits, and conduct problems. In addition, the relation between narcissism and conduct problems was moderated by self-esteem level, such that children with relatively high levels of narcissism and low self-esteem showed the highest rates of conduct-problem symptoms. PMID- 12573940 TI - Social Skills Training in Children With Attention Deficit Hyperactivity Disorder: A Randomized-Controlled Clinical Trial. AB - Evaluated efficacy of social skills training (SST) on children with 2 subtypes of attention deficit hyperactivity disorder (ADHD). Participants were 120 children (30 girls, 90 boys), ages 8 to 12 with ADHD-Inattentive type (ADHD-I; n = 59) or Combined type (ADHD-C; n = 61). The children were randomly assigned within diagnosis subtype to the treatment condition (8 weeks of SST) or the no intervention control condition. SST led to greater improvements in both parent- and child-perceived assertion skills in the children with ADHD, yet did not affect the other domains of social competence. Diagnostically heterogeneous groups led to greater improvements on parent-report of their child's cooperation and assertion abilities as well as children's report of their own empathy skills. Diagnostically homogeneous groups led to greater decreases in externalizing behaviors at posttreatment but not at follow-up. Children with comorbid oppositional defiant disorder (ODD) did not benefit as much from the intervention. Children with ADHD-I improved in assertion skills more than children with ADHD-C, yet the 2 diagnostic entities did not differ in improvement levels across all other social skills. PMID- 12573941 TI - A prospective test of the association between hyperarousal and emotional numbing in youth with a history of traumatic stress. AB - Investigated the hypothesis that emotional numbing may develop as a result of hyperarousal using a prospective design. Forty-two children between the ages of 7 and 14 with a history of trauma and posttraumatic stress disorder (PTSD) symptoms were assessed with the Clinician-Administered PTSD Scale for Children and Adolescents (CAPS-CA; Nader et al., 1996) and were reassessed 1 year later. Results indicated that hyperarousal symptoms were concurrently positively correlated with emotional numbing at both time points. Moreover, Time 1 hyperarousal symptoms were associated with emotional numbing at Time 2 and predicted Time 2 emotional numbing even when controlling for each of the other symptom clusters of PTSD at Time 1 as well as other concurrent (i.e., Time 2) PTSD symptoms. Results are discussed in terms of the implications of the findings for enhancing the understanding of PTSD symptoms in youth. PMID- 12573944 TI - Evolution of toxicology for risk assessment. AB - The science of toxicology has served society well in protecting public health and the environment. Governments, the industrial sector, and the public have relied on toxicology as the foundation to assess risks to both human and ecological populations from environmental factors, including chemicals, biologic agents, physical agents, and other stressors. To maintain its prominence, the science and practice of toxicology will need to embrace the revolution underway in biology. Systems biology and biotechnologies derived from sequencing of the human genome, referred to as "genomics," have created exciting possibilities for application to human health and environmental risk assessment. Yet this rapid advance of science and technology can be overshadowed by inconsistency in study design and sampling strategies; by the lack of quantitative or qualitative correlations of exposure, dose, or adverse effects; and by the lack of bioinformatics tools and analytical methods necessary to manage the volume of research findings. These limitations may render results uninterpretable and difficult, if not impossible, to use in risk assessment. Recommendations will be discussed to improve integrating systems biology and genomics into risk assessment so that the inherent promise of these new approaches can be realized. PMID- 12573945 TI - Short-term repeated-dose toxicity profile of archaeosomes administered to mice via intravenous and oral routes. AB - Archaeosomes, liposomes made from polar ether lipids of archaea, show promise for vaccine and drug delivery applications. The potential toxicity of intravenously (14, 70, or 140 mg/kg/day for 5 consecutive days) and orally (gavaged at 55, 275, or 550 mg/kg/day for 10 consecutive days) administered unilamellar archaeosomes, prepared from the total polar lipids (TPLs) extracted from several species of archaea, was assessed in female BALB/c mice. Liposomes prepared from an ester phospholipid composition were included for comparative purposes. Control groups of mice were administered 0.1 ml phosphate-buffered saline (PBS) by either route. Animals were monitored at least once daily for temperature, body weight, and clinical signs of adverse reactions. One day after the last dose, the mice were sacrificed. Blood was collected for selected biochemical/enzyme analyses, and the major organs (heart, lungs, liver, spleen, kidneys) were weighed and examined macroscopically. In addition, the spleens were examined histologically. At the two lower dosages of intravenously administered vesicles, there were no significant indications of toxicity, as compared with the PBS-administered control group. At the highest intravenous dose of 140 mg/kg/day, archaeosomes prepared from the TPL of the extreme halophiles, Halobacterium salinarum and Natronobacterium magadii, indicated potential toxicity, as evidenced by clinical signs (hyperactivity and/or piloerection), drop in body temperature, and loss in body weight. Spleens from mice administered some archaeosomes types, primarily at the highest intravenous dose tested, were enlarged, had increased organ weight, and microscopic examination revealed mild to moderate expansion of the red pulp with increased numbers of hematopoietic cells, but no changes in the white pulp. There were similar clinical signs at one or more of the higher oral doses of the ester liposomes and some of the archaeosome types; however, no other apparent toxicity was observed. Based on this limited mouse study, archaeosomes were generally well tolerated after intravenous or oral delivery at the dosages so indicated in this study. PMID- 12573946 TI - Extent and timeliness of tissue repair determines the dose-related hepatotoxicity of chloroform. AB - As a part of mixture toxicity studies, the objective of the present investigation was to validate the hypothesis that the rate and extent of liver tissue repair response to a given dose determines the end result of toxicity (death or recovery), regardless of the mechanisms by which injury is inflicted, using a well-known environmental pollutant, chloroform (CHCl(3)). In future, the data will be used to compare with the results of mixtures containing CHCl(3) to aid in characterizing the safety of chemical mixtures and to construct a physiologically based pharmacokinetic (PBPK) model for dose, route, and species extrapolation. Hepatotoxicity and tissue repair were measured in male Sprague-Dawley rats (S-D) receiving a 10-fold dose range of CHCl(3) (74, 185, 370, and 740 mg/kg, IP) during a time course of 0 to 96 hours. Liver injury, as assessed by plasma alanine aminotransferase (ALT) and sorbitol dehydrogenase (SDH) elevation, increased with dose over the 10-fold dose range. Because CHCl(3) is also known to cause kidney damage, blood urea nitrogen (BUN) and creatinine were measured to evaluate the kidney injury. With doses up to 370 mg/kg, liver injury increased in a dose-related fashion, which peaked at 24 hours and returned to normal after 48 hours, whereas at highest dose (740 mg/kg), the injury was progressive resulting in 90% mortality. Blood and liver CHCl(3) levels were quantified using gas chromatography (GC) over a time course of 30 to 360 minutes. The dose-related increase in the blood and liver CHCl(3) levels were consistent with dose dependent liver injury. Tissue regeneration response, as measured by [(3)H] thymidine incorporation into hepatocellular nuclear DNA peaked at 36 hours in rats treated with the lower two doses of CHCl(3) (74 and 185 mg/kg). Further increase in CHCl(3) dose to 370 mg/kg resulted in an earlier increase in [(3)H] thymidine incorporation at 24 hours, which peaked at 36 hours. However, at the highest dose of CHCl(3) (740 mg/kg), tissue repair was delayed and attenuated, allowing for unrestrained progression of liver injury. The kidney injury markers after CHCl(3) administration were not different from controls. These results support the concept that in addition to the magnitude of tissue repair response, the time at which this response occurs is critical in restraining the progression of injury. Measuring tissue repair and injury as simultaneous biological responses to toxic agents might increase the usefulness of dose-response paradigms in predictive toxicology and risk assessment. Although the dosimetry of the present study was well beyond the environmental exposure levels of CHCl(3), a PBPK model will be developed in future based upon these data to evaluate the effects at environmental levels. PMID- 12573947 TI - Cis-urocanic acid increases immunotoxicity and lethality of dermally administered permethrin in C57BL/6N mice. AB - Immunomodulatory effects of a single topical permethrin exposure, 5-day exposure to cis-urocanic acid (cUCA), or a combination of the two chemicals were evaluated in 4- to 5-week-old female C57BL/6N mice. Permethrin alone decreased thymic weight and cellularity. Although cUCA alone did not affect thymic end points, coexposure to topical permethrin and cUCA exacerbated the thymolytic effects of permethrin. The single topical dose of permethrin also depressed several immune responses in isolated splenic leukocytes. This included splenic T-cell proliferative response to mitogen, splenic macrophage hydrogen peroxide production, and splenic B lymphocyte-specific antibody production. Unlike the effect of coexposure to these agents on thymic end points, cUCA did not exacerbate permethrin's adverse effect on any of the splenic end points examined. These results appear to suggest divergent mechanisms by which these compounds affect precursor and functionally mature T cells. At the doses used in this study, permethrin caused neurotoxic effects, including lethality, in a portion of the mice. For undetermined reasons, cUCA significantly increased the rate of lethality caused by permethrin. Although the permethrin doses used in this study exceed that typically used in human medicine, these results raise some concerns about the possibility that sunlight, via cUCA, may increase the risk of adverse central nervous system and immune effects caused by permethrin alone. PMID- 12573948 TI - The sensitizer 2,4-dinitrofluorobenzene activates caspase-3 and induces cell death in a skin dendritic cell line. AB - In this work, a dendritic cell line derived from mouse skin (FSDC) was used, as an in vitro experimental model, to evaluate the cytotoxic effect of two chemical sensitizers, a strong sensitizer (2,4-dinitrofluorobenzene, DNFB) and a weak sensitizer (2,4-dichloronitrobenzene, DCNB). The results indicated that DNFB reduces the cellular metabolism of FSDC, as evaluated by the reduction of the tetrazolium salt, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). All the DNFB concentrations tested, ranging from 5.2 micro M to 26 micro M, significantly inhibited the MTT reduction after 1 hour of cell exposure to the sensitizer. In contrast, incubation of FSDC with the weak sensitizer DCNB had no significant effect on the MTT reduction assay. When the cells were incubated with DNFB (13 micro M), for 3 and 6 hours, morphological changes characteristics of cell death by apoptosis were observed, as assessed by propidium iodide (PI) DNA staining and annexin-V externalization analysis. These results correlate well with an increase of caspase-3-like activity after FSDC exposure to DNFB (13 micro M) for 6 hours. Together, these results indicate that apoptotic death of skin dendritic cells occurs after exposure to the sensitizer DNFB, although necrotic cell death was also observed when the cells were incubated with high concentrations of DNFB (26 micro M), or after long periods of cell exposure to the chemical DNFB (13 micro M, for 6 hours). PMID- 12573949 TI - A standardized approach for iris color determination. AB - Latanoprost, the phenyl-substituted prostaglandin F2alpha, has been found to be an effective agent for glaucoma therapy. This prostaglandin derivative exerts ocular hypotensive activity but is also associated with an untoward side effect, namely iris color changes. Latanoprost provoked iris color changes in cynomolgus monkeys and in multicenter clinical trials. Until now photographs were taken and compared with color plates to document these changes. The disadvantage of this method is obvious, i.e., the color luminance varies between measurements due to changes in the developer. Furthermore, subjective comparison of color changes relative to color plates rendered judgment subject to impression and opinion rather than to objective data. Therefore, a computerized method using a 3-CCD video camera attached to a slit lamp was developed. The signals were transferred to a computer and a single frame, which was "frozen" by means of a "grabber card." Camera and the computer had previously been calibrated and color plates were measured to check the standard conditions. They were evaluated by a software program displaying average color (as red, green, and blue values) of the selected area. This method provides a fast and accurate way to quantify color changes in the iris of both experimental animals and clinical trials. PMID- 12573950 TI - Neurotoxicity and immunotoxicity assessment in CBA/J mice with chronic Toxoplasma gondii infection and single-dose exposure to methylmercury. AB - Toxoplasma gondii is a protozoan parasite that localizes in the brain where it can cause life-threatening disease. Methylmercury (MeHg) is a well-documented neurotoxicant that accumulates in the brain. We investigated end points associated with immunotoxicity and neurotoxicity in mice exposed to MeHg during a chronic T. gondii infection. Two groups of 6-week-old, female CBA/J mice were either fed 25 T. gondii tissue cysts of the ME-49 strain or given vehicle. Six weeks later, half of the mice in each group were orally gavaged with a single dose of 20 mg/kg body weight of MeHg, creating four groups of mice (vehicle control, T. gondii, MeHg, and T. gondii/MeHg). Mice were sacrificed 7 days post MeHg exposure. MeHg exposure caused a significant decrease in mouse body weight. MeHg administration resulted in an increase of splenic cellularity and spleen-to body weight ratios. MeHg had no significant effect on the percentages of CD4(+), CD8(+), or non-T-cell subpopulations in the spleen. MeHg dosed mice demonstrated an increase in absolute numbers of splenic CD4(+), CD8(+), or non-T cells when compared to mice in control and T. gondii-infected groups. Thymic CD4(+)CD8(+) T cell subpopulations were decreased (p <.05) by MeHg with or without a concurrent T. gondii infection. There was a significant (p <.05) increase in brain tissue cyst counts within the group exposed to both MeHg and T. gondii (16 +/- 4, mean +/- SE, n = 7) versus T. gondii alone (4 +/- 1, n = 8). Histopathological examination demonstrated encephalitis, gliosis, and meningitis in brains from mice infected with T. gondii. These data indicate that exposure to both MeHg and T. gondii has synergistic effects, with effects of MeHg especially on the immune system. PMID- 12573956 TI - Evaluation of dry and wet block cutting and recommendation for a masonry company. PMID- 12573957 TI - Worker exposure to 1,3,5-triglycidyl isocyanurate (TGIC) in powder paint coating operations. PMID- 12573958 TI - Use of dermal LD(50) as a criterion for skin notation. PMID- 12573962 TI - Risk factors for low back pain among Filipino manufacturing workers and their anthropometric measurements. AB - This study looked into the prevalence of and risk factors for low back pain among workers in manufacturing industries in the Philippines. Anthropometric measurements were also done to establish the design principles of the working equipment, protective equipment, and tools of the Filipino worker to prevent musculoskeletal disorder. This was a cross-sectional study using a stratified random sampling technique. Thirty-one industries were selected. Various workstations were sampled from each industry where subjects were selected. There were 495 workers surveyed for the symptoms questionnaire and 544 for the anthropometric measurements. Results showed that 5.1 percent experienced discomfort, 2 percent had trunk rigidity, and 1.4 percent had both limitations of trunk motion and activities of daily living. Logistic regression showed that low back pain was significantly associated with leaning, bending, and carrying for 2 8 hours (p at.05), and with standing for 2-8 hours (p at.001). It was also found to affect work performance and more likely to occur 14 times as often after work as during the initial work sessions. Anthropometric measurements of the workers showed that the mean height is 159.96 cm, mean chest height is 115.70 cm, waist height is 96.95 cm, and knee height is 47.37 cm. Anthropometric data between sexes was also presented. This anthropometry can be used for the design of working equipment of Filipino workers. It is recommended that risk factors for low back pain be understood and equipment be designed according to the body proportions of the workers identified in this study. PMID- 12573963 TI - Sound exposure among the Finnish National Opera personnel. AB - The purpose of the study was to determine how and when the personnel of the Finnish National Opera are exposed to noise and whether exposure depends on musical selection of repertoire. Additionally, an evaluation of sound exposure level due to individual rehearsals was included. The measurements were done using individual noise dosimeters and fixed-point measurements. From the measurements, annual noise exposure in the Opera was evaluated. The conductors, dancers, and double bass players were exposed to levels below 85 decibels, A-weighted, dB(A), which is the national action level. The choir members were exposed to sound levels of 92 and 94 dB(A). Within the orchestra, the highest sound exposure levels were found among percussionists, 95 dB(A); flute/piccolo players, 95 dB(A); and brass players, 92-94 dB(A). Other sound exposure levels among orchestra members varied from 83 to 89 dB(A). Soloists and rehearsal pianists are likely to be exposed to sound levels exceeding the national action level. From an exposure perspective, the individual rehearsals, 79-100 dB(A), proved to be as important as performances and group rehearsals, 82-99 dB(A), among orchestra musicians and choir singers. The ambient sound level for the lighting crew was 76 +/- 4 dB(A). However, the measured sound levels at the ear varied from 77 to 92 dB(A) due to the communication via headphones that had individual volume control. For the majority of personnel of the Finnish National Opera, sound exposure level exceeded the national action level value of 85 dB(A). Artists exceeded the action level during both individual and group rehearsals, as well as during performances. Hearing protection has been designed for musicians. Education/reinforcement is required to ensure it is worn. PMID- 12573961 TI - An evaluation of Fourier transform infrared (FTIR) spectroscopy for detecting organic solvents in expired breath. AB - The aim of this study was to test the performance of gas-phase FTIR analysis on human breath samples. Ten volatile organic compounds (VOC) were examined for applicability to FTIR spectroscopy (ethanol, ethylbenzene, n-hexane, methyl ethyl ketone, methyl tert-butyl ether, m-xylene, 1,1,1-trichloroethane, trichloroethylene, tetrachloroethylene, and toluene). Three sets of detection limits (LOD) were determined for comparison. LOD(1) were generated from partial least squares (PLS) calibration methods using spectroscopic software, LOD(2) from spiked breath samples, and LOD(3) from blank breath samples. Mixed expired breath samples from four subjects were spiked at varying levels with four different VOC (hexane, methyl ethyl ketone, m-xylene and 1,1,1-trichloroethane) to validate spectral data and test overall accuracy. Breath samples spiked with m-xylene also were validated by GC/FID analysis. PLS-derived LOD(1) ranged from 0.06-2.47 ppm. Spiked breath sample LOD(2) ranged from 0.52-1.21 ppm. Blank breath LOD(3) measurements ranged from 0.17-1.70 ppm, except for ethanol, which had an LOD of 11.2 ppm. Predicted concentrations for carbon dioxide (slope = 1.06), m-xylene (slopes = 1.19, 1.21), and methyl ethyl ketone (slope = 0.93) were fairly accurate, while concentrations were underpredicted for n-hexane (slope = 0.69) and 1,1,1-trichloroethane (slopes = 0.58-0.66). PMID- 12573964 TI - Particle concentration profile in a vertical displacement flow: a study in an industrial hall. AB - The effect of displacement flow on the distribution of aerosol concentration was investigated in an industrial hall. According to the displacement ventilation principle, vertical upflow is accomplished by introducing fresh air, cooler than room air, into the occupied zone near floor level. The fresh air is introduced from low-velocity devices and heated by warm processes. This technique allows warm air contaminants to rise to the ceiling, and the rising plume is then exhausted close to the ceiling. This study presents the results of a field study conducted in an industrial environment. The aerosol properties and behavior, especially the vertical gradients, are characterized in a displacement flow field. The results indicate that the fine particles, less than 1 microm in diameter, are transported away from the breathing zone by the ventilation process. However, the air quality is significantly influenced by the emission source, and therefore the number concentration of fine and ultrafine (smaller than 0.1 microm in diameter) aerosol particles in the breathing zone was clearly elevated compared to that of the incoming clean air. The vertical gradients displayed clear size dependence; the strongest gradients were found for particles between 0.003 and 0.015 microm in diameter. PMID- 12573965 TI - Extent of fungal growth on fiberglass duct liners with and without biocides under challenging environmental conditions. AB - Eight brands of fiberglass duct liners, including three that contained biocides, were exposed to challenging environmental conditions that would promote fungal growth. Twenty-four rectangular sheet metal ducts in three groups of eight ducts per group were lined with the eight selected liners. Each group of ducts was exposed to one of the three test conditions within an environmental chamber for a period of 15 days. These conditions were a) 75 percent RH, b) 75 percent RH plus water spray, c) 75 percent RH plus dry nutrient, and d) 75 percent RH plus water plus nutrient. Viable spores of Aspergillus niger were aerosolized into each duct as seed. On the 16th day, air and surface samples for fungal spores were collected from inside ducts. The results of air sampling using N6 sampler and visual inspection indicated that two out of three biocide-containing liners, Permacote and Toughgard, inhibited fungal growth but only under condition A. The third biocide-containing liner, Aeroflex Plus, was effective even when it was wet (conditions A and B). All three biocide-containing liners failed to inhibit fungal growth under conditions C and D. Among the five other types of liners that did not contain biocides, ATCO Flex with a smooth Mylar coating was more preferable, exhibiting lower fungal activity during conditions A, B, and C. All liners failed under condition D when nutrient and water were added together. Surface sampling using adhesive tape failed to produce representative results, apparently due to rough/porous surface of duct liners. It was concluded that duct liners with biocide treatment could be less promoting to microbial growth under high humidity as long as their surfaces remain clean and water-free. A liner with an impermeable and smooth surface seems to be less subject to microbial growth under most conditions than biocide-containing liners having porous and/or rough surfaces. PMID- 12573966 TI - Estimating methyl bromide exposure due to offgassing from fumigated commodities. AB - Methyl bromide (MB) is used to fumigate diverse commodities. During fumigation, the commodity can sorb a substantial mass of MB, which does not chemically react and is termed a residue. During subsequent commodity handling, the residue offgasses and can lead to MB inhalation exposure among processing workers. Although MB has a low 1 ppm 8-hr TLV-TWA as recommended by the American Conference of Governmental Industrial Hygienists (ACGIH(R)) and is considered a potential occupational carcinogen by the National Institute for Occupational Safety and Health, the recent industrial hygiene literature contains no pertinent exposure data. Limited measurements made in 1992 by the California Department of Pesticide Regulation are summarized here, but associated information on exposure determinants is lacking. In this article, mathematical models are used to integrate data on MB residue offgassing with several processing scenarios to estimate potential exposure levels. The main finding is that if a large volume of commodity rapidly offgasses MB and is handled under conditions of low ventilation, the potential exists for MB exposures above the 1 ppm TLV-TWA value. However, the combination of handling a smaller commodity volume and less rapid offgassing may be the more typical scenario. It is recommended that a pilot study be conducted to measure current MB exposure levels, test the validity of the mathematical models, and collect industry-wide data on exposure determinants. By using the latter data as inputs for validated models, public health scientists could estimate the distribution of MB exposure levels across the commodity processing industry. PMID- 12573976 TI - Improved erectile function after Rho-kinase inhibition in a rat castrate model of erectile dysfunction. AB - Androgens are reported to act as strong modulators of erectile function influencing both nitric oxide and vasoconstrictor signaling. Castration results in a depressed erectile response that is associated with a loss of nitric oxide production and increased responsiveness to constrictive agents. The increased vasoconstrictor response may be a result of an active RhoA/Rho-kinase signaling pathway. We report here results of studies designed to test the hypothesis that inhibition of the Rho-kinase pathway restores erectile function in a castrate model by relaxing the smooth muscle. Mean arterial (MAP) and corpus cavernosal (CCP) pressures were monitored during intracavernosal injection of the Rho-kinase inhibitor Y-27632. Castration reduced the maximal erectile response (CCP/MAP) by 33%, and testosterone replacement restored the response (intact, 0.736 +/- 0.040; castrate, 0.492 +/- 0.022; testosterone, 0.681 +/- 0.073). Injection of Y-27632 increased CCP in all experimental groups; it also left shifted the voltage response curve and increased the maximal CCP/MAP response (intact, 0.753 +/- 0.091; castrate, 0.782 +/- 0.081; testosterone treated, 0.894 +/- 0.033). Y-27632 dose dependently relaxed phenylephrine-stimulated cavernosal tissues. Cavernosal tissues showed increased RhoA and Rho-kinase protein levels after castration. Our data support the hypothesis that an active Rho/Rho-kinase pathway contributes to the reduced erectile response after castration due to an upregulation of RhoA/Rho kinase protein levels and that inhibition of this pathway may serve as an effective treatment for erectile dysfunction. PMID- 12573977 TI - Guanylyl cyclase mediates ANP-induced vasoconstriction of murine splenic vessels. AB - We have previously shown that ANP causes differential constriction of the splenic vasculature of the rat (veins greater than arteries), which may be inhibited by blocking the production of cGMP with A7195. In this paper, we report experiments done on vessels derived from guanylyl cyclase (GC)-A knockout mice. Small splenic arteries ( approximately 150-microm diameter) and veins ( approximately 250 microm diameter) were dissected from male GC-A-deficient 129sv mice or age matched wild-type controls and mounted in a wire myograph. In the wild-type mice, ANP exhibited higher potency in the veins than in the arteries (EC(50) values wild-type mice: artery, 8 +/- 3 x 10(-9) M, n = 5 vs. vein, 6 +/- 4 x 10(-10) M, n = 5; P < 0.05). The concentration-response curve for ANP-induced vasoconstriction was also shifted leftward in denuded compared with intact arteries (EC(50) values: denuded artery: 5 +/- 3 x 10(-10) M, n = 5 vs. intact artery, 8 +/- 3 x 10(-9) M, n = 5; P < 0.05), i.e., the denuded vessels were more reactive. By contrast, ANP caused no significant change in tension from baseline in intact splenic arteries, intact splenic veins, or denuded splenic arteries derived from the GC-A-deficient mice, although these vessels did show normal concentration-dependent increases in tension to phenylephrine. We conclude that ANP causes vasoconstriction in the splenic vasculature by an endothelium independent mechanism, mediated via guanylyl cyclase. PMID- 12573978 TI - Postnatal intracerebroventricular exposure to neuropeptide Y causes weight loss in female adult rats. AB - We investigated the effect of repetitive postnatal (2-7 days) intracerebroventricular administration of neuropeptide Y (NPY) on food intake and body weight gain in the 3- to 120-day-old Sprague-Dawley rats. NPY caused a 32% transient increase in body weight gain with elevated circulating insulin concentrations within 24 h. This early intervention led to the persistence of hyperinsulinemia and relative hyperleptinemia with euglycemia in the 120-day-old female alone. This perturbation was associated with 50% suppression in adult female hypothalamic NPY concentrations and a 50-85% decline in NPY immunoreactivity in the paraventricular and arcuate nuclei. This change was paralleled by a approximately 20% decline in food intake and body weight gain at 60 and 120 days. However, when exogenous NPY was stereotaxically reinjected into the paraventricular nucleus of the approximately 120-day-old adult females who were pretreated with NPY postnatally, an increase in food intake and body weight gain was noted, attesting to no disruption in the NPY end-organ responsivity. We conclude that postnatal intracerebroventricular NPY has long-lasting effects that predetermine the resultant adult phenotype in a sex-specific manner. PMID- 12573979 TI - Cardiac effects of hypocretin-1 in nucleus ambiguus. AB - Although recent studies have reported hypocretin 1 (hcrt-1)-like-immunoreactivity (ir) within the region of the nucleus ambiguus (Amb) in the caudal brain stem, the function of hcrt-1 in the Amb on cardiovascular function is not known. Three series of experiments were done in male Wistar rats to investigate the effects of microinjections of hcrt-1 into Amb on heart rate (HR), mean arterial pressure (MAP), and the arterial baroreceptor reflex. In the first series, a detailed mapping of the distribution of hcrt-1- and hcrt-1 receptor (hcrtR-1)-like-ir was obtained of the Amb region. Although hcrt-1-like- and hcrtR-1-like-ir were found throughout the rostrocaudal extent of the Amb and adjacent ventrolateral medullary reticular formation, most of the hcrtR-1-like-ir was observed in the area just ventral to the compact formation of Amb, in the region of the external formation of the nucleus (Ambe). In the second series, the Amb region that contained hcrt-1 and hcrtR-1-ir was explored for sites that elicited changes in HR and MAP in urethane and alpha-chloralose-anesthetized rats. Microinjections of hcrt-1 (0.5-2.5 pmol) into the Ambe elicited a dose-related decrease in HR, with little or no direct change in MAP. The small decreases in MAP were found to be secondary to the HR changes. The largest bradycardia responses were elicited from sites in the Ambe. Administration (iv) of the muscarinic receptor antagonist atropine methyl bromide or ipsilateral vagotomy abolished the HR response, indicating that the HR response was due to activation of vagal cardiomotor neurons. In the final series, microinjections of hcrt-1 into the Ambe significantly potentiated the reflex bradycardia elicited by activation of the baroreflex as a result of the increased MAP after the intravenous injection of phenylephrine. These data suggest that hcrt-1 in the Ambe activates neuronal systems that alter the excitability of central circuits that reflexly control the circulation through the activation of vagal preganglionic cardioinhibitory neurons. PMID- 12573980 TI - Involvement of sympathetic efferents but not capsaicin-sensitive afferents in nociceptin-mediated dual control of rat synovial blood flow. AB - This study set out to examine the vasomotor effects of the opioid-like peptide nociceptin on knee joint capsular blood flow in urethane-anaesthetized rats. Topical application of nociceptin (10(-15)-10(-8) mol) caused a progressive fall in joint perfusion that was significantly inhibited by the specific nociceptin receptor antagonist [Phe(1)-(CH(2)-NH)-Gly(2)] Nociceptin(1-13)-NH(2) as well as the nonspecific opioid antagonist naloxone. To test whether this constrictor response was sympathetically mediated, we administered nociceptin in animals treated with guanethidine to produce sympathetic blockade or in the presence of the alpha-adrenoceptor antagonist phentolamine. Both guanethidine treatment and phentolamine coadministration attenuated the constrictor response to nociceptin. Inhibition of nociceptin-mediated vasoconstriction revealed a supplementary hyperemic response that persisted in animals whose knee joints were treated with 1% capsaicin to destroy the articular unmyelinated nerve supply. These results show that, in the rat knee, peripheral administration of nociceptin primarily causes a sympathetically mediated vasoconstriction. In addition, high-dose nociceptin produces a vasodilatatory response that is likely due to the direct action of nociceptin on vascular smooth muscle and not by a neurogenic mechanism. PMID- 12573981 TI - Uniformity in dynamic baroreflex regulation of left and right cardiac sympathetic nerve activities. AB - Functional laterality of cardiac sympathetic nerve stimulation in chronotropic and inotropic effects is well known. Whether left (LSNA) and right (RSNA) cardiac sympathetic nerve activities show laterality during dynamic baroreflex activation remains to be determined. In nine anesthetized, vagotomized, and aortic denervated rabbits, we randomly perturbed intracarotid sinus pressure (CSP) in both carotid sinus regions while simultaneously recording LSNA and RSNA. The baroreflex neural arc transfer function from CSP to LSNA and from CSP to RSNA revealed derivative characteristics, i.e., the magnitude of LSNA and RSNA responses became greater as the input frequency of CSP perturbation increased. The average slope of increasing gain in the frequencies between 0.03 and 0.3 Hz showed no difference between LSNA and RSNA responses (9.7 +/- 2.9 vs. 9.7 +/- 3.1 dB/decade, means +/- SD). The amplitude ratio and phase difference between LSNA and RSNA approximated unity and zero radians, respectively, in the frequencies from 0.01 to 1 Hz. In addition, the LSNA-RSNA relationship during stepwise CSP perturbation from 40 to 160 mmHg showed a straight line (r(2) ranged from 0.969 to 0.999). These findings indicate no laterality in the dynamic as well as static baroreflex regulation of LSNA and RSNA as far as grouped axonal activity is concerned. PMID- 12573982 TI - Circadian phase-shifting effects of nocturnal exercise in older compared with young adults. AB - Exercise can phase shift the circadian rhythms of young adults if performed at the right time of day. Similar research has not been done in older adults. This study examined the circadian phase-delaying effects of a single 3-h bout of low intensity nocturnal exercise in older (n = 8; 55-73 yr old) vs. young (n = 8; 20 32 yr old) adults. The exercise occurred at the beginning of each subject's habitual sleep time, and subjects sat in a chair in dim light during the corresponding time in the control condition. The dim-light melatonin onset (DLMO) was used as the circadian phase marker. The DLMO phase delayed more after the exercise than after the control condition. On average, the difference in phase shift between the exercise and control conditions was similar for older and young subjects, demonstrating that the phase-shifting effects of exercise on the circadian system are preserved in older adults. Therefore, exercise may potentially be a useful treatment to help adjust circadian rhythms in older and young adults. PMID- 12573983 TI - Protein kinase C modulates pulmonary endothelial permeability: a paradigm for acute lung injury. AB - The intracellular serine/threonine kinase protein kinase C (PKC) has an important role in the genesis of pulmonary edema. This review discusses the PKC-mediated mechanisms that participate in the pulmonary endothelial response to agents involved in lung injury characteristic of the respiratory distress syndrome. Thus the paradigms of PKC-induced lung injury are discussed within the context of pulmonary transvascular fluid exchange. We focus on the signal transduction pathways that are modulated by PKC and their effect on lung endothelial permeability. Specifically, alpha-thrombin, tumor necrosis factor (TNF)-alpha, and reactive oxygen species are discussed because of their well-established roles in both human and experimental lung injury. We conclude that PKC, most likely PKC alpha, is a primary supporter for lung endothelial injury in response to alpha thrombin, TNF-alpha, and reactive oxygen species. PMID- 12573984 TI - Regulation of nitric oxide production in limb and ventilatory muscles during chronic exercise training. AB - In this study, we evaluated the differential influence of chronic treadmill training (30 m/min, 15% incline, 1 h/day, 5 days/wk) on nitric oxide (NO) production and NO synthase (NOS) isoform expression as well as 3-nitrotyrosine formation (footprint of peroxynitrite) both in limb (gastrocnemius) and ventilatory (diaphragm) muscles. A group of exercise-trained rats and a control group (no training) were examined after a 4-wk experimental period. Exercise training elicited an approximate fourfold rise in gastrocnemius NOS activity and augmented protein expression of the endothelial (eNOS) and neuronal (nNOS) isoforms of NOS to approximately 480% and 240%, respectively. Qualitatively similar but quantitatively smaller elevations in NOS activity and eNOS and nNOS expression were observed in the diaphragm. No detectable inducible NOS (iNOS) protein expression was found in any of the muscle samples. Training increased the intensity of 3-nitrotyrosine only in the gastrocnemius muscle. We conclude that whole body exercise training enhances both limb and ventilatory muscle NO production and that constitutive and not iNOS isoforms are responsible for increased protein tyrosine nitration in trained limb muscles. PMID- 12573985 TI - Absorption of intact albumin across rat alveolar epithelial cell monolayers. AB - Transport characteristics of intact albumin were investigated using primary cultured rat alveolar epithelial cell monolayers. The apical-to-basolateral (ab) flux of intact fluorescein isothiocyanate (FITC)-labeled albumin (F-Alb) is greater than basolateral-to-apical (ba) flux at the same upstream [F-Alb]. Net absorption of intact F-Alb occurs with half-maximal concentration of approximately 1.6 microM and maximal transport rate of approximately 0.15 fmol.cm(-2).s(-1). At 15 and 4 degrees C, both ab and ba F-Alb fluxes are not different from zero, collapsing net absorption. The presence of excess unlabeled albumin (but not other macromolecule species) in either the apical or basolateral fluid significantly reduces both ab and ba unidirectional F-Alb fluxes. Photoaffinity labeling of apical cell membranes revealed an approximately 60-kDa protein that exhibits specificity for albumin. These data indicate that net absorption of intact albumin takes place via saturable receptor-mediated transcellular endocytotic processes recognizing albumin, but not other macromolecules, that may play an important role in alveolar homeostasis in the mammalian lung. PMID- 12573986 TI - Surfactant strengthens the inhibitory effect of C-reactive protein on human lung macrophage cytokine release. AB - In this study we investigated the effect of acute-phase levels of C-reactive protein (CRP) on cytokine production by pulmonary macrophages in the presence or absence of pulmonary surfactant. Both human alveolar and interstitial macrophages as well as human surfactant were obtained from multiple organ donor lungs. Precultured macrophages were stimulated with LPS alone or together with IFN-gamma in the presence or absence of CRP, surfactant, and combinations. Releases of TNF alpha and of IL-1beta to the medium were determined. We found that CRP could modulate lung inflammation in humans by decreasing the production of proinflammatory cytokines by both alveolar and interstitial macrophages stimulated with LPS alone or together with IFN-gamma. The potential interaction between CRP and surfactant phospholipids did not overcome the effect of either CRP or surfactant on TNF-alpha and IL-1beta release by lung macrophages. On the contrary, CRP and pulmonary surfactant together had a greater inhibitory effect than either alone on the release of proinflammatory cytokines by lung macrophages. PMID- 12573987 TI - An enhancer region determines hSP-B gene expression in bronchiolar and ATII epithelial cells in transgenic mice. AB - Regulation of the surfactant protein B gene (SP-B) is developmentally controlled and highly tissue specific. To elucidate the SP-B gene temporal/spatial expression pattern in lung development at the transcriptional level, a transgenic mouse model line carrying the human SP-B (hSP-B) 1.5-kb 5'-flanking regulatory region and the lacZ gene was established. Expression of hSP-B 1.5-kb lacZ gene started at the onset of lung formation [embryonic day 9 (E9)] and was restricted to epithelial cells throughout prenatal and postnatal lung development. In the adult lung, hSP-B 1.5-kb lacZ gene expression was restricted to bronchiolar and alveolar type II epithelial cells. In lung explant culturing studies, the hSP-B 1.5-kb lacZ gene was highly expressed in newly formed epithelial tubules during the respiratory branching process. In a second transgenic mouse line, an enhancer region, which binds to thyroid transcription factor-1, retinoic acid receptor, signal transducers and activators of transcription 3, and nuclear receptor coactivators (SRC-1, ACTR, TIF2, and CBP/p300), was deleted from the hSP-B 1.5-kb lacZ gene. The deletion abolished hSP-B lacZ gene expression in bronchiolar epithelial cells and significantly reduced its expression level in alveolar type II epithelial cells in transgenic mice. PMID- 12573988 TI - Bleomycin-induced apoptosis of alveolar epithelial cells requires angiotensin synthesis de novo. AB - Primary cultures of rat type II alveolar epithelial cells (AECs) or human AEC derived A549 cells, when exposed to bleomycin (Bleo), exhibited concentration dependent apoptosis detected by altered nuclear morphology, fragmentation of DNA, activation of caspase-3, and net cell loss over time. In both cell culture models, exposure to Bleo caused time-dependent increases in angiotensinogen (ANGEN) mRNA. Antisense oligonucleotides against ANGEN mRNA inhibited Bleo induced apoptosis of rat AEC or A549 cells by 83 and 84%, respectively (P < 0.01 and P < 0.05), and prevented Bleo-induced net cell loss. Apoptosis of rat AECs or A549 cells in response to Bleo was inhibited 91% by the ANG-converting enzyme inhibitor captopril or 82%, respectively, by neutralizing antibodies specific for ANG II (both P < 0.01). Antagonists of ANG receptor AT(1) (losartan, L-158809, or saralasin), but not an AT(2)-selective blocker (PD-123319), inhibited Bleo induced apoptosis of either rat AECs (79%, P < 0.01) or A549 cells (83%, P < 0.01) and also reduced the activity of caspase-3 by 52% (P < 0.05). These data indicate that Bleo, like Fas(L) or TNF-alpha, induces transactivation of ANG synthesis de novo that is required for AEC apoptosis. They also support the theory that ANG system antagonists have potential for the blockade of AEC apoptosis in situ. PMID- 12573990 TI - Intratracheal gene transfer of decorin reduces subpleural fibroproliferation induced by bleomycin. AB - Decorin, a small leucin-rich proteoglycan, is a negative regulator of transforming growth factor-beta, but the antifibrotic effect of decorin gene transfer has not been examined in a mouse model of usual interstitial pneumonia (UIP). We constructed a replication-defective recombinant adenovirus harboring human decorin gene (AdCMV.DC) and administered 1 x l0(9) plaque-forming units of AdCMV.DC intratracheally or intravenously to C57BL/6 mice with intraperitoneal injection of bleomycin, which induces a subpleural fibroproliferation, mimicking UIP, by day 28. Only intratracheal administration of AdCMV.DC increased decorin mRNA expression in the lung and decreased the hydroxyproline content augmented in bleomycin-induced pulmonary fibrosis (1.13 +/- 0.02 to 0.96 +/- 0.02, P = 0.006). In contrast, intravenous administration of AdCMV.DC increased the decorin expression only in the liver, but not in the lung, and without reducing lung fibrosis. These results indicate that adenoviral decorin gene transfer is effective only by direct administration to fibrosing lungs. PMID- 12573989 TI - Intrauterine hypertension decreases lung VEGF expression and VEGF inhibition causes pulmonary hypertension in the ovine fetus. AB - Although vascular endothelial growth factor (VEGF) plays a vital role in lung vascular growth in the embryo, its role in maintaining endothelial function and modulating vascular structure during late fetal life has not been studied. We hypothesized that impaired lung VEGF signaling causes pulmonary hypertension, endothelial dysfunction, and structural remodeling before birth. To determine whether lung VEGF expression is decreased in an experimental model of persistent pulmonary hypertension of the newborn (PPHN), we measured lung VEGF and VEGF receptor protein content from fetal lambs 7-10 days after ductus arteriosus ligation (132-140 days gestation; term = 147 days). In contrast with the surge in lung VEGF expression during late gestation in controls, chronic intrauterine pulmonary hypertension reduced lung VEGF expression by 78%. To determine whether VEGF inhibition during late gestation causes pulmonary hypertension, we treated fetal lambs with EYE001, an aptamer that specifically inhibits VEGF(165). Compared with vehicle controls, EYE001 treatment elevated pulmonary artery pressure and pulmonary vascular resistance by 22 and 50%, respectively, caused right ventricular hypertrophy, and increased wall thickness of small pulmonary arteries. EYE001 treatment reduced lung endothelial nitric oxide synthase protein content by 50% and preferentially impaired the pulmonary vasodilator response to ACh, an endothelium-dependent agent. We conclude that chronic intrauterine pulmonary hypertension markedly decreases lung VEGF expression and that selective inhibition of VEGF(165) mimics the structural and physiological changes of experimental PPHN. We speculate that hypertension downregulates VEGF expression in the developing lung and that impaired VEGF signaling may contribute to the pathogenesis of PPHN. PMID- 12573991 TI - Histone acetylation regulates epithelial IL-8 release mediated by oxidative stress from environmental particles. AB - Increases in the levels of environmental particulate matter with a diameter of <10 microm diameter (PM(10)) in the air are associated with a variety of adverse health effects, particularly chronic lung and cardiovascular diseases. The expression of many inflammatory genes involves the remodeling of the chromatin structure provided by histone proteins. Histone acetylation causes the unwinding of chromatin structure, therefore allowing transcription factor access to promoter sites. Acetylation is reversible and is regulated by histone acetyltransferases (HATs), which promote acetylation, and deacetylases, which promote deacetylation. PM(10) and H(2)O(2) increased IL-8 protein release from A549 cells after 24-h treatment, and this was enhanced by histone deacetylase inhibition by trichostatin A (cotreatment). PM(10) and H(2)O(2) treatment also increased HAT activity as well as the level of acetylated histone 4 (H4). PM(10) enhanced H4 acetylation that was mediated by oxidative stress as shown by thiol antioxidant inhibition. Acetylation of H4 mediated by PM(10) was associated with the promoter region of the IL-8 gene. These data suggest that remodeling of chromatin by histone acetylation plays a role in PM(10)-mediated responses in the lungs. PMID- 12573992 TI - Systemic and regional hemodynamic effects of angiotensin-(1-7) in rats. AB - The systemic and regional hemodynamics effects of ANG-(1-7) were examined in urethane-anesthetized rats. The blood flow distribution (kidneys, skin, mesentery, lungs, spleen, brain, muscle, and adrenals), cardiac output, and total peripheral resistance were investigated by using fluorescent microspheres. Blood pressure and heart rate were recorded from the brachial artery. ANG-(1-7) infusion (110 fmol x min(-1) x 10 min(-1) iv) significantly increased blood flow to the kidney (5.10 +/- 1.07 to 8.30 +/- 0.97 ml x min(-1) x g(-1)), mesentery (0.73 +/- 0.16 to 1.17 +/- 0.49 ml x min(-1) x g(-1)), brain (1.32 +/- 0.44 to 2.18 +/- 0.85 ml x min(-1) x g(-1)), and skin (0.07 +/- 0.02 to 0.18 +/- 0.07 ml x min(-1) x g(-1)) and the vascular conductance in these organs. ANG-(1-7) also produced a significant increase in cardiac index (30%) and a decrease in total peripheral resistance (2.90 +/- 0.55 to 2.15 +/- 0.28 mmHg x ml(-1) x min x 100 g). Blood flow to the spleen, muscle, lungs, and adrenals, as well as the blood pressure and heart rate, were not altered by the ANG-(1-7) infusion. The selective ANG-(1-7) antagonist A-779 reduced the blood flow in renal, cerebral, mesenteric, and cutaneous beds and blocked the ANG-(1-7)-induced vasodilatation in the kidney, mesentery, and skin, suggesting a significant role of endogenous ANG-(1-7) in these territories. The effects of ANG-(1-7) on the cerebral blood flow, cardiac index, systolic volume, and total peripheral resistance were partially attenuated by A-779. A high dose of ANG-(1-7) (11 pmol x min(-1) x 10 min(-1)) caused an opposite effect of that produced by the low dose. Our results show for the first time that ANG-(1-7) has a previously unsuspected potent effect in the blood flow distribution and systemic hemodynamics. PMID- 12573993 TI - Development of electrical activity in cardiac myocyte aggregates derived from mouse embryonic stem cells. AB - Embryonic stem cells differentiate into cardiac myocytes, repeating in vitro the structural and molecular changes associated with cardiac development. Currently, it is not clear whether the electrophysiological properties of the multicellular cardiac structure follow cardiac maturation as well. In long-term recordings of extracellular field potentials with microelectrode arrays consisting of 60 substrate-integrated electrodes, we examined the electrophysiological properties during the ongoing differentiation process. The beating frequency of the growing preparations increased from 1 to 5 Hz concomitant to a decrease of the action potential duration and action potential rise time. A developmental increase of the conduction velocity could be attributed to an increased expression of connexin43 gap junction channels. Whereas isoprenalin elicited a positive chronotropic response from the first day of spontaneous beating onward, a concentration-dependent negative chronotropic effect of carbachol only developed after approximately 4 days. The in vitro development of the three-dimensional cardiac preparation thus closely follows the development described for the mouse embryonic heart, making it an ideal model to monitor the differentiation of electrical activity in embryonic cardiomyocytes. PMID- 12573994 TI - Isoflurane activates rat mitochondrial ATP-sensitive K+ channels reconstituted in lipid bilayers. AB - Activation of mitochondrial ATP-sensitive K(+) (mitoK(ATP)) channels is critical in myocardial protection induced by preconditioning with volatile anesthetics or brief periods of ischemia. In this study, we characterized rat mitoK(ATP) channels reconstituted in lipid bilayers and examined their direct regulation by isoflurane. Mitochondria and the inner membrane fraction were isolated from rat ventricles and fused into lipid bilayers. On the basis of their inhibition by 5 hydroxydecanoate (5-HD)/ATP or activation by diazoxide, mitoK(ATP) channels of several conductance states were observed in symmetrical (150 mM) potassium glutamate (26, 47, 66, 83, and 105 pS). Isoflurane (0.8 mM) increased the cumulative open probability from 0.09 +/- 0.02 at baseline to 0.50 +/- 0.09 (P < 0.05, n = 5), which was inhibited by 5-HD. Isoflurane caused a dose-dependent rightward shift in ATP inhibition of mitoK(ATP) channels, which increased the IC(50) for ATP from 335 +/- 4 to 940 +/- 34 microM at 0.8 mM (P < 0.05, n = 5 approximately 8). We conclude that direct activation of the mitoK(ATP) channel by isoflurane is likely to contribute to volatile anesthetic-induced myocardial preconditioning. PMID- 12573995 TI - Integrin shedding as a mechanism of cellular adaptation during cardiac growth. AB - Integrin-mediated cell-extracellular matrix (ECM) interactions are essential for multiple cellular processes; however, little is known regarding integrin turnover during these events. Recent studies have demonstrated shedding of cell surface molecules and suggested this as a potential mechanism for integrin turnover. Confocal microscopy of mouse hearts under different physiological conditions demonstrated the presence of beta(1)-integrin-immunoreactive material in the interstitium. Culture media from neonatal rat cardiac myocytes and fibroblasts contained a 55-kDa fragment of beta(1)-integrin. Attachment to ECM components, response to phorbol 12-myristate 13-acetate stimulation, and matrix metalloproteinase inhibition assays demonstrated that fibroblasts responded differently to the fragment compared with myocytes. The beta(1)-integrin fragment stimulated myocyte attachment to collagen and the fragment itself bound a variety of ECM proteins. These studies indicate that as myocytes and fibroblasts change size and shape, cellular contacts with the ECM are altered, resulting in the liberation of a beta(1)-integrin fragment from the cell surface. Integrin shedding may represent a novel mechanism of rapidly modifying cell-ECM contacts during various cellular processes. PMID- 12573996 TI - Infarct-induced chronic heart failure increases bidirectional protein movement across the alveolocapillary barrier. AB - Chronic heart failure (CHF) is associated with adaptive structural changes at the alveolocapillary barrier that may be associated with altered protein permeability. Bidirectional protein movement across the barrier was studied in anesthetized rats with infarct-induced CHF by following (125)I-labeled albumin ((125)I-albumin) flux into the alveoli and the leakage of surfactant protein (SP) B from the alveoli into the circulation. Three groups were studied: controls [0% left ventricular (LV) infarction], moderate infarct (25-45% LV infarction), and large infarct (>46% LV infarction). Wet and dry lung weights increased in the large infarct group (both P < 0.001), consistent with increased lung water and solid lung tissue. (125)I-albumin flux increased across the endothelial (P < 0.001) and epithelial (P < 0.01) components of the alveolocapillary barrier in the large infarct group. Plasma SP-B increased 23% with moderate infarcts (P < 0.05) and 97% with large infarcts (P < 0.001), independent of alveolar levels. Lavage fluid immune cells (P < 0.01) and myeloperoxidase activity (P < 0.05) increased in the large infarct group, consistent with inflammation. Bidirectional protein movement across the alveolocapillary barrier is increased in CHF, and alveolar inflammation may contribute to this pathophysiological defect. PMID- 12573997 TI - Unloading-induced remodeling in the normal and hypertrophic left ventricle. AB - To date, no study has assessed the degree of similarity between left ventricular (LV) reverse remodeling and atrophic remodeling. Stable LV hypertrophy was induced by creation of an arteriovenous fistula (AVF) in Lewis rats (32 days). LV unloading was induced by heterotopic transplantation of normal (NL-HT) and/or hypertrophic (AVF-HT) hearts (7 days). We compared indexes of remodeling in AVF, NL-HT, and AVF-HT groups with those of normal controls. LV unloading induced decreases in cardiomyocyte size in NL-HT and AVF-HT hearts. NL-HT and AVF-HT LV were both characterized by relative increases in collagen concentration that were largely a reflection of decreases in myocyte volume. NL-HT and AVF-HT LV were associated with similar increases in matrix metalloproteinase (MMP-2 and -9) zymographic activity, without change in the abundance of the tissue inhibitors of the MMPs. In contrast, AVF-HT, but not NL-HT, was associated with a dramatic increase in collagen cross-linking. Our findings suggest an overall similarity in the response of the normal and hypertrophic LV to surgical unloading. However, the dramatic increase in collagen cross-linking after just 1 wk of unloading suggests a potential difference in the dynamics of collagen metabolism between the two models. Further studies will be required to determine the precise molecular mechanisms responsible for these differences in extracellular matrix regulation. However, with respect to these and related issues, heterotopic transplantation of hypertrophied hearts will be a useful small animal model for defining mechanisms of myocyte-matrix interactions during decreased loading conditions. PMID- 12573998 TI - Structural response of microcirculatory networks to changes in demand: information transfer by shear stress. AB - Matching blood flow to metabolic demand in terminal vascular beds involves coordinated changes in diameters of vessels along flow pathways, requiring upstream and downstream transfer of information on local conditions. Here, the role of information transfer mechanisms in structural adaptation of microvascular networks after a small change in capillary oxygen demand was studied using a theoretical model. The model includes diameter adaptation and information transfer via vascular reactions to wall shear stress, transmural pressure, and oxygen levels. Information transfer is additionally effected by conduction along vessel walls and by convection of metabolites. The model permits selective blocking of information transfer mechanisms. Six networks, based on in vivo data, were considered. With information transfer, increases in network conductance and capillary oxygen supply were amplified by factors of 4.9 +/- 0.2 and 9.4 +/- 1.1 (means +/- SE), relative to increases when information transfer was blocked. Information transfer by flow coupling alone, in which increased shear stress triggers vascular enlargement, gave amplifications of 4.0 +/- 0.3 and 4.9 +/- 0.5. Other information transfer mechanisms acting alone gave amplifications below 1.6. Thus shear-stress-mediated flow coupling is the main mechanism for the structural adjustment of feeding and draining vessel diameters to small changes in capillary oxygen demand. PMID- 12573999 TI - Gender dimorphic tissue perfusion response after acute hemorrhage and resuscitation: role of vascular endothelial cell function. AB - Proestrous female rodents are protected from the deleterious effects of trauma hemorrhage that are observed in males. We hypothesized that the gender dimorphic outcome after trauma-hemorrhage might be related to gender differences in endothelial function and organ perfusion under such conditions. Male and cycle matched proestrous female Sprague-Dawley rats underwent a midline laparotomy, hemorrhagic shock (40 mmHg for approximately 90 min), and resuscitation (Ringer lactate, 4x shed blood volume over 60 min). Various parameters were measured 2 h after completion of resuscitation. In the first set of animals, the left ventricle was cannulated and heart performance (maximal rate of left ventricular pressure increase) as well as cardiac output and organ perfusion rates were determined with (85)Sr microspheres. In the second set of animals, aortic vessel rings were harvested and relaxation in response to acetylcholine and nitroglycerin was measured. In the third set of animals, in situ isolated small intestine was perfused to measure the response of the splanchnic vessel bed to acetylcholine and nitroglycerin. After trauma-hemorrhage and resuscitation, females maintained cardiac output and demonstrated increased splanchnic and cardiac perfusion compared with males. Moreover, female intestines did not manifest the endothelial dysfunction that was observed in male intestines after hemorrhagic shock. We conclude that proestrous females show improved endothelial function and tissue perfusion patterns after hemorrhagic shock and that this gender-specific response might be a potential mechanism contributing to the beneficial effects of the proestrus stage under such conditions. PMID- 12574000 TI - Ventricular remodeling induced by retinoic acid supplementation in adult rats. AB - Retinoic acid (RA) plays a role in regulating cardiac geometry and function throughout life. The aim of this study was to analyze the cardiac effects of RA in adult rats. Wistar rats were randomly allocated to a control group (n = 18) receiving standard rat chow and a group treated with RA (n = 14) receiving standard rat chow supplemented with RA for 90 days. All animals were evaluated by echocardiography, isolated papillary muscle function, and morphological studies. Whereas the RA-treated group developed an increase in both left ventricular (LV) mass and LV end-diastolic diameter, the ratio of LV wall thickness to LV end diastolic diameter remained unchanged when compared with the control group. In the isolated papillary muscle preparation, RA treatment decreased the time to peak developed tension and increased the maximum velocity of isometric relengthening, indicating that systolic and diastolic function was improved. Although RA treatment produced an increase in myocyte cross-sectional area, the myocardial collagen volume fraction was similar to controls. Thus our study demonstrates that small physiological doses of RA induce ventricular remodeling resembling compensated volume-overload hypertrophy in rats. PMID- 12574001 TI - Catecholamines act via a beta-adrenergic receptor to maintain fetal heart rate and survival. AB - Mice lacking catecholamines die before birth, some with cardiovascular abnormalities. To investigate the role of catecholamines in development, embryonic day 12.5 (E12.5) fetuses were cultured and heart rate monitored. Under optimal oxygenation, wild-type and catecholamine-deficient fetuses had the same initial heart rate (200-220 beats/min), which decreased by 15% in wild-type fetuses during 50 min of culture. During the same culture period, catecholamine deficient fetuses dropped their heart rate by 35%. Hypoxia reduced heart rate of wild-type fetuses by 35-40% in culture and by 20% in utero, assessed by echocardiography. However, catecholamine-deficient fetuses exhibited greater hypoxia-induced bradycardia, reducing their heart rate by 70-75% in culture. Isoproterenol, a beta-adrenergic receptor (beta-AR) agonist, reversed this extreme bradycardia, restoring the rate of catecholamine-deficient fetuses to that of nonmutant siblings. Moreover, isoproterenol rescued 100% of catecholamine deficient pups to birth in a dose-dependent, stereo-specific manner when administered in the dam's drinking water. An alpha-AR agonist was without effect. When wild-type fetuses were cultured with adrenoreceptor antagonists to create pharmacological nulls, blockade of alpha-ARs with 10 microM phentolamine or beta ARs with 10 microM bupranolol alone or in combination did not reduce heart rate under optimal oxygenation. However, when combined with hypoxia, beta-AR blockade reduced heart rate by 35%. In contrast, the muscarinic blocker atropine and the alpha-AR antagonist phentolamine had no effect. These data suggest that beta-ARs mediate survival in vivo and regulate heart rate in culture. We hypothesize that norepinephrine, acting through beta-ARs, maintains fetal heart rate during periods of transient hypoxia that occur throughout gestation, and that catecholamine-deficient fetuses die because they cannot withstand hypoxia-induced bradycardia. PMID- 12574002 TI - Recruitment of smooth muscle cells and arterial vasomotion. AB - Investigating the recruitment and synchronization of smooth muscle cells (SMCs) is the key to understanding the physical mechanisms leading to contraction and spontaneous diameter oscillations of arteries, called vasomotion. We improved a method that allows the correlation of calcium oscillations (flashing) of individual SMCs with mean calcium variations and arterial contraction using confocal microscopy. Endothelium-stripped rat mesenteric arteries were cut open, loaded with dual calcium fluorescence probes, and stimulated by increasing concentrations of the vasoconstrictors phenylephrine (PE) and KCl. We found that the number and synchronization of flashing cells depends on vasoconstrictor concentration. At low vasoconstrictor concentration, few cells flash asynchronously and no local contraction is detected. At medium concentration, recruitment of cells is complete and synchronous, leading to strip contraction after KCl stimulation and to vasomotion after PE stimulation. High concentration of PE leads to synchronous calcium oscillations and fully contracted vessels, whereas high concentration of KCl leads to a sustained nonoscillating increase of calcium and to fully contracted vessels. We conclude that the number of simultaneously recruited cells is an important factor in controlling rat mesenteric artery contraction and vasomotion. PMID- 12574003 TI - Postsystolic shortening of ischemic myocardium: a mechanism of abnormal intraventricular filling. AB - Acute myocardial ischemia has been associated with abnormal filling patterns in the left ventricular (LV) apex. We hypothesized that this may in part be due to postsystolic shortening of ischemic apical segments, which leads to reversal of early diastolic apical flow. Fourteen open-chest anesthetized dogs were instrumented with micromanometers in the LV apex and left atrium and myocardial sonomicrometers in the anterior apical LV wall. Intraventricular filling by color Doppler and wall motion by strain Doppler echocardiography (SDE) were assessed from an apical view. Measurements were taken before and after 5 min of left anterior descending coronary artery (LAD) occlusion. In four dogs, we measured the pressure difference between the LV apex and outflow tract. At baseline, peak early diastolic flow velocities in the distal one-third of the LV were directed toward apex (9.2 +/- 1.6 cm/s). After LAD occlusion, the velocities reversed ( 2.3 +/- 0.4 cm/s, P < 0.01), indicating that blood was ejected from the apex toward the base during early filling. This interpretation was confirmed by wall motion analysis, which showed postsystolic shortening of apical myocardial segments. The postsystolic shortening represented 9.7 +/- 1.7% (P < 0.01) and 14.2 +/- 2.4% (P < 0.01) of end-diastolic segment length by SDE and sonomicrometry, respectively. Consistent with the velocity changes, we found reversal of the early diastolic pressure gradient from the LV apex to outflow tract. In the present model, acute LAD occlusion resulted in reversal of early diastolic apical flow, and this was attributed to postsystolic shortening of dyskinetic apical segments. The clinical diagnostic importance of this finding remains to be determined. PMID- 12574004 TI - Changes in amino-terminal portion of human B2 receptor selectively increase efficacy of synthetic ligand HOE 140 but not of cognate ligand bradykinin. AB - Recently, we have shown that a widely used antagonist of the human bradykinin B(2) receptor (B(2)R) HOE 140 acts as a full agonist of the chicken ornithokinin receptor (B(o)R). To understand the molecular mechanisms underlying differential efficacy of HOE 140 for the various kinin receptors, we have constructed chimeric kinin receptors (CKR) in which the amino-terminal portion including the first two transmembrane regions and the first extracellular loop (CKR-2) or only the second transmembrane region and the first extracellular loop (CKR-1) of B(2)R were substituted with the corresponding segments of B(o)R. Ligand efficacy of synthetic ligand HOE 140 decreased in the order B(o)R > CKR-2 > CKR-1 > B(2)R, whereas the efficacy of the endogenous kinin ligand was unchanged. Enhanced HOE 140 efficacy was not due to a structural change in the ligand binding site or to an enhanced receptor expression level. Rather, heterologous binding competition studies indicated that structural change(s) introduced into the engineered receptors caused a selective reduction in apparent affinity of HOE 140 for the uncoupled inactive receptor state R but not for the active G protein-coupled state R*, thereby increasing the ratio of R* over R for a given ligand concentration. Our results may help explain the unusually broad efficacy spectrum of HOE 140, which varies from inverse to full agonism, depending on kinin receptor subtype, tissue origin, or species. PMID- 12574005 TI - Enalapril attenuates endothelin-1-induced hypertension via increased kinin survival. AB - Recent studies have shown that angiotensin-converting enzyme (ACE) inhibitors attenuate endothelin-1 (ET-1)-induced hypertension, but the mechanisms for this effect have not been clarified. Initial experiments were conducted to contrast the effect of the ACE inhibitor enalapril, the combined ACE-neutral endopeptidase inhibitor omapatrilat, and the angiotensin II receptor antagonist candesartan on the hypertensive and renal response to ET-1 in anesthetized Sprague-Dawley rats. Acute intravenous infusion of ET-1 (10 pmol x kg(-1) x min(-1)) for 60 min significantly increased mean arterial pressure (MAP) from 125 +/- 8 to 145 +/- 8 mmHg (P < 0.05) and significantly decreased glomerular filtration rate (GFR) from 0.31 +/- 0.09 to 0.13 +/- 0.05 ml x min(-1) x 100 g kidney wt(-1). Pretreatment with enalapril (10 mg/kg iv) before ET-1 infusion inhibited the increase in MAP (121 +/- 4 vs. 126 +/- 4 mmHg) before and during ET-1 infusion, respectively (P < 0.05) without blocking the effect of ET-1 on GFR. In contrast, neither omapatrilat (30 mg/kg) nor candesartan (10 mg/kg) had any effect on ET-1-induced increases in MAP or decreases in GFR. To determine whether the effect of enalapril was due to the decrease in angiotensin II or increase in kinin formation, rats were given REF-000359 (1 mg/kg iv), a selective B(2) receptor antagonist, with or without enalapril before ET-1 infusion. REF-000359 completely blocked the effect of enalapril on ET-1 infusion (MAP was 117 +/- 5 vs. 135 +/- 5 mmHg before and during ET-1 infusion, respectively, P < 0.05). REF-000359 alone had no effect on the response to ET-1 infusion (MAP was 117 +/- 4 vs. 144 +/- 4 mmHg before and during ET-1 infusion, respectively, P < 0.05). REF-000359 with or without enalapril had no significant effect on the ability of ET-1 infusion to decrease GFR. These findings support the hypothesis that decreased catabolism of bradykinin and its subsequent vasodilator activity oppose the actions of ET-1 to increase MAP. PMID- 12574006 TI - Amiodarone attenuates fluoride-induced hyperkalemia in vitro. AB - Poisoning by hydrofluoric acid or fluoride salts results in hypocalcemia, hypomagnesemia, and hyperkalemia with subsequent cardiac dysrhythmias. In previous studies, quinidine attenuated fluoride-induced hyperkalemia in vitro, and enhanced survival in animals. Like quinidine, amiodarone is a potassium channel blocker, although amiodarone is more familiar to clinicians due to its recent inclusion in advanced cardiac life support (ACLS) protocols. OBJECTIVES: This in-vitro study of human erythrocytes was designed to determine whether amiodarone could attenuate fluoride-induced hyperkalemia. METHODS: Six healthy volunteers each donated 60 mL of blood on three occasions. Each specimen was divided into 12 tubes, incubated at 37 degrees C, and oxygenated with room air. An aqueous sodium fluoride (F(-)) solution was added to tubes 1-9. Incremental amounts of quinidine were added to tubes 1-4 (Q(1)-Q(4)) to attain calculated concentrations of 0.73 microg/mL, 1.45 microg/mL, 2.9 microg/mL, and 5.8 microg/mL, respectively. Incremental amounts of amiodarone were added to tubes 5 8 (A(1)-A(4)) to attain calculated concentrations of 0.38 microg/mL, 0.75 microg/mL, 1.5 microg/mL, and 3.0 microg/mL, respectively. Tubes 9-12 were controls for each of F(-), amiodarone, quinidine alone, and no additive, respectively. Extracellular potassium concentration ([K(+)]) was followed, and an objective endpoint was defined as the rise in potassium concentration at 6 hours. RESULTS: Fluoride produced a significant change in [K(+)] by 6 hours in all samples. Quinidine produced a J-shaped curve in its ability to attenuate the rise in [K(+)], with only one concentration, Q(3), demonstrating significance versus tube 9 (control). Amiodarone also demonstrated a J-shaped dose-response effect, with statistical significance at A(1), A(2), and A(3) versus tube 9 (control). There was no significant difference among the effective concentrations (Q(3), A(1), A(2), and A(3)) of both drugs. CONCLUSIONS: In this in-vitro model using human blood, amiodarone and quinidine both attenuated F(-)-induced hyperkalemia. Further study is indicated to determine whether amiodarone enhances survival in F(-)-poisoned animals. PMID- 12574007 TI - Tissue adhesives for traumatic lacerations: a systematic review of randomized controlled trials. AB - OBJECTIVE: To summarize the best available evidence for the effect of tissue adhesives (TAs) in managing traumatic lacerations in children and adults. METHODS: A search was conducted using the Cochrane Controlled Trials Register, Medline, and EMBase for relevant studies. The authors also searched the citations of selected studies and contacted relevant authors and manufacturers. The search included randomized controlled trials (RCTs) comparing a TA versus standard wound closure (sutures, staples, adhesive strips) (SWC) or two TAs for acute, linear, low-tension, traumatic lacerations in an emergency or primary care setting. Data were extracted by one reviewer and checked for accuracy by a second reviewer. Two reviewers independently assessed masked copies for quality. Outcomes of cosmesis (subgroups of age, wound location, and need for deep sutures), pain, procedure time, ease of use, and complications were analyzed separately for two comparisons: 1). TA versus SWC; and 2). TA versus TA. RESULTS: Eight studies compared a TA with SWC. No significant difference was found for cosmesis at any time point examined. Subgroup analysis was possible only for age; no significant difference was found. Pain scores [Parent Visual Analogue Scale weighted mean difference (VAS WMD) = -15.7 mm; 95% CI = -21.9 to -9.5] and procedure time (WMD = -5.7 minutes; 95% CI = -8.2 to -3.1) significantly favored TAs. Small, statistically significant risk differences were found for dehiscence [favoring SWC; number needed to harm (NNH) = 25; 95% CI = 14 to 100] and erythema (favoring TAs; NNH = 8; 95% CI = 4 to 100). Only one study compared two TAs (butylcyanoacrylate versus octylcyanoacrylate) for pediatric facial lacerations. No significant difference was found for cosmesis, pain, procedure time, or complications. CONCLUSIONS: TAs are an acceptable alternative to SWC for simple traumatic lacerations. No difference in cosmesis was found between TAs and SWC, or between different TAs. Tissue adhesives offer the benefits of decreased procedure time and less pain, compared with SWC. A small increased rate of dehiscence with TAs must be considered when choosing the closure method (NNH = 25). PMID- 12574008 TI - Beta-human chorionic gonadotropin levels and the likelihood of ectopic pregnancy in emergency department patients with abdominal pain or vaginal bleeding. AB - Strategies for diagnosing ectopic pregnancy that defer endovaginal ultrasound in women with suggestive symptoms and serum beta-human chorionic gonadotropin (beta hCG) levels less than 1500 mIU/mL ignore the increased risk of ectopic pregnancy in these patients. OBJECTIVE: To quantify this increased risk by establishing and comparing the beta-hCG distributions of symptomatic women with ectopic pregnancies, abnormal intrauterine pregnancies, and normal intrauterine pregnancies. METHODS: The authors reviewed the records of a cohort of women who visited an urban emergency department (ED) during a 34-month period with abdominal pain or vaginal bleeding and non-zero quantitative beta-hCG levels. Explicit criteria were used to determine whether the pregnancy ultimately turned out to be intrauterine and normal, intrauterine and abnormal, or ectopic. Probability distributions were compared using frequency distributions, receiver operating characteristic (ROC) curves, and likelihood ratios. RESULTS: Of 730 ED patients included in the analysis, 96 (13%) had ectopic pregnancies, 253 (35%) had abnormal intrauterine pregnancies, and 381 (52%) had normal intrauterine pregnancies. The beta-hCG distributions of patients with ectopic pregnancies and abnormal intrauterine pregnancies were similar and much lower than the beta-hCG distribution of patients with normal intrauterine pregnancies. A beta-hCG level less than 1500 mIU/mL more than doubled the odds of ectopic pregnancy (likelihood ratio = 2.24). Of the 158 patients with beta-hCG below 1500 mIU/mL, 40 (25%; 95% confidence interval [CI] = 19% to 32%) had ectopic pregnancies, and only 25 (16%; CI = 11% to 22%) had normal intrauterine pregnancies. CONCLUSIONS: In women with pain or bleeding and serum beta-hCG levels less than 1500 mIU/mL, the risk of ectopic pregnancy is substantially increased, while the likelihood of normal intrauterine pregnancy is low. PMID- 12574009 TI - The effect of hospital occupancy on emergency department length of stay and patient disposition. AB - Emergency department (ED) overcrowding is a common problem. Despite a widespread belief that low hospital bed availability contributes to ED overcrowding, there are few data demonstrating this effect. OBJECTIVES: To identify the effect of hospital occupancy on ED length of stay for admitted patients and patient disposition. METHODS: This was an observational study design using administrative data at a 500-bed acute care teaching hospital. All patients presenting to the ED between April 1993 and June 1999 were included in the study. The predictor variable was daily hospital occupancy. Outcome measures included daily ED length of stay for admitted patients, daily consultation rate, and daily admission rate. The models controlled for the average daily age of ED patients and the average daily "arrival density" index, which adjusts for patient volume and clustering of patient arrivals. RESULTS: The average hospital occupancy was 89.7%. On average 155 patients visited the ED daily; 21% were referred to hospital physicians and 19% were admitted. The median ED length of stay for admitted patients was 5 hours 54 minutes (interquartile range 5 hr 12 min to 6 hr 42 min). Daily ED length of stay for admitted patients increased 18 minutes (95% CI = 12 to 24) when there was an absolute increase in occupancy of 10%. The ED length of stay appeared to increase extensively when hospital occupancy exceeded a threshold of 90%. Consultation and admission rates were not influenced by hospital occupancy. CONCLUSIONS: Increased hospital occupancy is strongly associated with ED length of stay for admitted patients. Increasing hospital bed availability might reduce ED overcrowding. PMID- 12574010 TI - Etomidate for rapid-sequence intubation in young children: hemodynamic effects and adverse events. AB - OBJECTIVES: Physicians commonly use etomidate for adult rapid-sequence intubation (RSI), but the manufacturer does not recommend its use for children under 10 years of age due to a lack of data. The authors present their experience with etomidate for pediatric RSI in order to further develop its risk-benefit profile in this age group. METHODS: Trained abstractors reviewed the medical records for all children under 10 years old who received etomidate for RSI between July 1996 and April 2001. RESULTS: 105 children, with an average age of 3 (+/-2.9) years, received a median dose of 0.32 (+/-0.12) mg/kg of etomidate. The systolic blood pressure increased an average of 4 mm Hg (95% CI = -3.3 to 9.2); the diastolic blood pressure increased 7 mm Hg (95% CI = -3.1 to 11) within 10 minutes of receiving etomidate. The heart rate increased an average of 10 beats/min (95% CI = 4.0 to 17.4). Complications included three patients who vomited within 10 minutes of etomidate administration. There were no cases of documented myoclonus, status epilepticus, or new-onset seizures. Thirty-eight patients received corticosteroids during the hospital course, none for suspected adrenal insufficiency. Three patients died, all from severe brain injury. CONCLUSIONS: In children less than 10 years old, etomidate seems to produce minimal hemodynamic changes, and appears to have a low risk of clinically important adrenal insufficiency, myoclonus, and status epilepticus. The association between etomidate and emesis (observed in less than 3% of enrolled patients) remains unclear. For clinical situations in which minimal blood pressure changes during RSI are critical, etomidate appears to have a favorable risk-benefit profile for children under 10 years old. PMID- 12574011 TI - Ventilatory response during dissociative sedation in children-a pilot study. AB - OBJECTIVES: It is unclear whether ketamine induces subclinical respiratory depression when administered in dissociative doses intravenously (IV). The authors report a pilot study of capnography in emergency department (ED) pediatric patients receiving ketamine alone for procedural sedation, and describe serial measures of ventilatory response [end-tidal carbon dioxide (EtCO(2)), respiratory rate, pulse oximetry]. METHODS: The authors performed continuous capnography on a convenience sample of 20 ED pediatric patients who received ketamine 1.5 mg/kg IV for procedural sedation. RESULTS: Continuous EtCO(2) and pulse oximetry remained essentially unchanged following ketamine injection, and no EtCO(2) levels > 47 mm Hg were noted at any point throughout sedation. CONCLUSIONS: No hypoventilation was observed in 20 ED pediatric patients receiving ketamine 1.5 mg/kg administered IV over 1 minute. The authors found no evidence of respiratory depressant properties for this dissociative agent. PMID- 12574012 TI - Can emergency department triage nurses appropriately utilize the Ottawa Knee Rules to order radiographs?-An implementation trial. AB - OBJECTIVE: To determine whether triage nurses can successfully interpret the Ottawa Knee Rule (OKR) and order knee radiographs according to the OKR. METHODS: This was a prospective implementation trial of a clinical decision rule, set in a suburban, community emergency department (ED), evaluating a convenience sample of ED patients aged > 17 years with acute knee injuries. Patients were excluded for altered mental status, distracting injuries, and knee lacerations. Triage nurses and attending emergency physicians (EPs) were trained in appropriate use of the OKR. The triage nurses evaluated eligible patients and radiographs were ordered according to their interpretation of the OKR. EPs who were initially blinded to the triage assessments also evaluated the patients. EPs could add an x-ray order if, according to their assessment of the OKR, one was indicated and a radiograph had not been ordered by the nurse. Nurses and EPs recorded their blinded assessments on standardized data collection instruments. Kappa values were calculated to assess interobserver agreement (IOA) between nurses and EPs; sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) were calculated as appropriate. RESULTS: One hundred three patients were enrolled; 53% were female; 10 fractures were identified (9.7%). The IOAs between the nurses and EPs for each of the criteria were moderate to almost perfect: age-0.94; fibular head tenderness-0.4; isolated patellar tenderness 0.68; inability to bend knee to 90 degrees-0.73; inability to bear weight-0.76. The IOA was moderate (0.52) for the overall interpretation of the OKR by nurses and EPs. Sensitivity of nurse interpretation of the OKR for fracture was 70%, specificity 33%, NPV 91%, PPV 10%. Sensitivity of EP interpretation of the OKR for fracture was 100%, specificity 25%, NPV 100%, PPV 13%. CONCLUSIONS: Triage nurses showed fair to good ability to appropriately apply the OKR to pre-order knee radiographs. PMID- 12574013 TI - Incidence of traumatic lumbar puncture. AB - OBJECTIVE: To determine the incidence of traumatic lumbar puncture (LP). METHODS: A retrospective study was conducted at an urban, university tertiary care referral center with 50000 annual emergency department (ED) visits. The study population included all patients who had cerebrospinal fluid (CSF) samples sent to the laboratory between August 15, 2000, and August 14, 2001. The numbers of red blood cells (RBCs) recorded in the first and last CSF tubes, the location where the LP was performed, and the discharge summary and the discharge diagnoses from the particular visit were obtained. All patients with intracranial pathology and CSF obtained via neurosurgical procedure or fluoroscopic guidance were excluded from the study group. Given no clear definition of traumatic LP in the literature, the incidence of traumatic LP was calculated using a cutoff of greater than 400 RBCs (visual threshold for bloody fluid) and 1000 RBCs (arbitrary threshold selected by other authors) in CSF tube 1. Proportions were compared using chi-square statistics. RESULTS: Seven hundred eighty-six CSF samples were recorded over one year. Twenty-four samples were obtained from patients with intracranial pathology or were obtained via a neurosurgical procedure. Of the remaining 762 CSF samples in the study population, 119 (15.6%) were traumatic using a cutoff of 400 RBCs, and 80 (10.5%) were traumatic, using a cutoff of 1000 RBCs in tube 1. Five hundred three LPs were done in the ED and 259 were attributed to all other locations in the hospital. Using a cutoff of 400 RBCs, the incidence of traumatic LP in the ED was 13.3%, compared with 20% in the rest of the hospital (p < 0.025). Similarly, using a cutoff of 1000 RBCs, the incidence of traumatic LP in the ED was 8.9%, compared with 13.5% in the rest of the hospital (p = 0.1). The incidence of "champagne taps" (defined as zero RBCs in the first and last tubes) in the ED was 34.4%, compared with 24.3% in the rest of the hospital (p < 0.01). CONCLUSIONS: The incidence of traumatic lumbar puncture is approximately 15% using a cutoff of 400 RBCs and 10% using a cutoff of 1000 RBCs. In this study, the rate of traumatic lumbar puncture was significantly less (with a cutoff of 400 RBCs) and the rate of champagne tap was significantly greater for LPs done in the ED compared with the rest of the hospital. PMID- 12574014 TI - A survey of the emergency department population and their interest in preventive health education. AB - OBJECTIVE: To determine which preventive health information the emergency department (ED) population (patients and visitors) would be most interested in having available to them while they spend time in the waiting area. METHODS: This was a prospective survey of consecutive adults seated in the ED waiting area during a representative week on predetermined shifts. The survey asked them to indicate whether they would be interested in obtaining information about the following preventive health issues: breast cancer, prostate cancer, smoking, obesity, stress reduction, exercise programs, alcohol/drugs, HIV, blood pressure screening, immunizations, referrals to primary care physicians, Pap smears, car safety, smoke detectors, domestic and youth violence, depression, gun safety, and safe sex. RESULTS: Of the 1284 subjects approached, 878 (68%) made up the study group (56% female, mean age = 44 years, 60% white); 406 refused. The information people were most interested in obtaining was the following: 52% of the respondents were interested in referral to stress reduction programs, 51% in information about exercise programs, 42% in blood pressure screening, 40% in information about breast cancer screening, 33% in depression information/screening, 33% in prostate cancer screening, 26% in immunization against pneumococcus, 24% in immunization against tetanus, 26% in smoking cessation programs, and 26% in safe driving information. Women were most interested in breast cancer screening (64%); and men, in prostate cancer screening (55%). CONCLUSIONS: Of the 878 subjects in the study group, 96% were interested in obtaining information about one or more preventive health issues. An opportunity exists to respond to this interest by providing material for public health education in the waiting area of EDs. PMID- 12574015 TI - Creating the model of a clinical practice: the case of emergency medicine. AB - Medical schools and specialty societies have struggled to define a core content for medical students and practitioners but, to date, have been stymied by both political considerations and the sheer burden of the innumerable decisions that must be made to define the essence of a medical specialty. Six professional organizations representing the field of emergency medicine recently collaborated with the National Board of Medical Examiners to accomplish this objective by developing a Model of the Clinical Practice of Emergency Medicine. This document will provide support for the development of medical school and residency curricula, training program accreditation standards, board certification test specifications, and organizational agendas for postgraduate education, research, and patient advocacy for the specialty of emergency medicine. The authors present a description of the model and the process that was used to develop it with the belief that other medical disciplines that face similar issues and challenges could benefit from a similar undertaking. PMID- 12574016 TI - Emergency medicine leadership in industry-sponsored clinical trials. AB - OBJECTIVE: To identify and characterize emergency medicine (EM) researchers who, since 1990, have served on a steering committee (SC) or as overall principal investigator (PI) of an industry-sponsored, multicenter clinical trial involving a pharmaceutical or device. METHODS: North American EM research directors (RDs) and other prominent EM investigators (for those hospitals without a RD) were identified from eight sources, including the Society for Academic Emergency Medicine RD Interest Group and the Multicenter Airway Research Collaboration (MARC) database. The identified investigators were sent a screening survey requesting information regarding industry-sponsored clinical research at their site. The individual EM investigators identified by this screening survey were then interviewed by telephone (validation survey) to further explore their leadership experience in industry-sponsored clinical trials. RESULTS: Of 153 identified RDs and prominent EM researchers, 138 responded to the screening survey (90% response rate). Eighty-five EM investigators reportedly had served on a SC or as overall PI for an industry-sponsored clinical trial. Of these 85 North American EM investigators, 77 were available for a structured telephone interview (91% response rate). Although 41 (53%) of the investigators confirmed their leadership role, 36 (47%) had not served in either role. Among the 41 confirmed investigators, 19 (25%) had served as a SC member, 10 (13%) had served as overall PI, and 12 (16%) had experience in both roles. Individual responses provided suggestions for pursuing such leadership positions. CONCLUSIONS: These data suggest the opportunity to expand EM leadership in industry-sponsored clinical trials and demonstrate the need for validation of reports obtained by a departmental research contact. The suggestions from EM researchers who have attained these leadership roles may provide strategies for investigators interested in pursuing these positions. PMID- 12574017 TI - Can the bispectral index monitor quantify altered level of consciousness in emergency department patients? AB - OBJECTIVES: A daily part of emergency medicine practice includes assessing patients with altered levels of consciousness (ALOC). The authors hypothesized that a bispectral index monitor (BIS), a processed electroencephalographic monitor traditionally used to monitor patients under anesthesia, would represent an objective quantification of impairment of consciousness. They compared the BIS score with the Glasgow Coma Scale score (GCS) in emergency department (ED) patients with ALOC. METHODS: The authors performed a convenience sampling of ED adults presenting with ALOC (GCS 150 scans (p < 0.005). The presence of an ultrasound rotation at an emergency medicine residency program also produced a statistically significant increase in test score (OR 1.82; 95% CI = 1.29 to 2.55). Residents at programs spending the least time (6 to 15 hours) on didactic education throughout the residency predicted examination failure (OR 0.60; 95% CI = 0.39 to 0.93). Increasing the amount of resident ultrasound teaching by emergency physicians improved the score, but this did not reach significance (p = 0.357). CONCLUSIONS: Improved resident performance on an ultrasound written examination was associated with increasing resident year, number of scans performed, and the presence of an ultrasound rotation at the residency program. Increasing the number of didactic hours spent on ultrasound each year beyond 15 hours showed no improvement in resident performance. PMID- 12574019 TI - Child protection--lessons from Victoria Climbie. PMID- 12574020 TI - Medical experts and the criminal courts. PMID- 12574021 TI - Thyroid function tests and hypothyroidism. PMID- 12574024 TI - Bush criticised for not giving AIDS money to Global Fund. PMID- 12574022 TI - The prevention and treatment of jet lag. PMID- 12574025 TI - Researchers launch major initiative to hunt human cancer genes. PMID- 12574026 TI - Arkansas bans selling of clean urine to beat drug testing. PMID- 12574027 TI - "Weak" safety culture behind errors, says chief medical officer. PMID- 12574029 TI - Women still have difficulty getting local contraceptive advice. PMID- 12574033 TI - Chocolate cigarettes "recruit" children to smoking. PMID- 12574030 TI - Doctors told to use positive language in managing pain. PMID- 12574035 TI - Report says 100 Roma women have been forcibly sterilised in Slovakia. PMID- 12574040 TI - Sally Clark freed after appeal court quashes her convictions. PMID- 12574041 TI - GP suspended for enrolling patients in drug trials without consent. PMID- 12574042 TI - Managing communication with young people who have a potentially life threatening chronic illness: qualitative study of patients and parents. AB - OBJECTIVES: To examine young people's and parents' accounts of communication about cancer in childhood. DESIGN: Semistructured interviews analysed using the constant comparative method. SETTING: Paediatric oncology unit. PARTICIPANTS: 13 families, comprising 19 parents (13 mothers, six fathers) and 13 patients aged 8 17 years, recruited from one paediatric oncology unit. The patients had cancer or brain tumour. RESULTS: Most parents described acting in an executive-like capacity, managing what and how their children were told about their illness, particularly at the time of diagnosis. Their accounts were shaped by concerns to manage their identity as strong and optimistic parents and to protect their child's wellbeing. The patients identified elements of their parents' role that both facilitated and constrained their communication, and while they welcomed their parents' involvement, some expressed unease with the constraining aspects of their parents' role. Some young people described feeling marginalised in consultations and pointed to difficulties they experienced in encounters with some doctors. CONCLUSIONS: There are difficulties in managing communication with young people who have a chronic, life threatening illness. Health professionals need to be aware of how the social positioning of young people (relative to adults) and the executive role of parents can contribute to the marginalisation of young people and hamper the development of successful relationships between themselves and young patients. PMID- 12574043 TI - Risk of prevalent HIV infection associated with incarceration among injecting drug users in Bangkok, Thailand: case-control study. AB - OBJECTIVES: To identify risks for HIV infection related to incarceration among injecting drug users in Bangkok, Thailand. DESIGN: Case-control study of sexual and parenteral exposures occurring before, during, and after the most recent incarceration. SETTING: Metropolitan Bangkok. PARTICIPANTS: Non-prison based injecting drug users formerly incarcerated for at least six months in the previous five years, with documented HIV serostatus since their most recent release; 175 HIV positive cases and 172 HIV negative controls from methadone clinics. MAIN OUTCOME MEASURE: Injection of heroin and methamphetamine, sharing of needles, sexual behaviour, and tattooing before, during, and after incarceration. RESULTS: In the month before incarceration cases were more likely than controls to have injected methamphetamine and to have borrowed needles. More cases than controls reported using drugs (60% v 45%; P=0.005) and sharing needles (50% v 31%; P<0.01) in the holding cell before incarceration. Independent risk factors for prevalent HIV infection included injection of methamphetamine before detention (adjusted odds ratio 3.3, 95% confidence interval 1.01 to 10.7), sharing needles in the holding cell (1.9, 1.2 to 3.0), being tattooed while in prison (2.1, 1.3 to 3.4), and borrowing needles after release (2.5, 1.3 to 4.4). CONCLUSIONS: Injecting drug users in Bangkok are at significantly increased risk of HIV infection through sharing needles with multiple partners while in holding cells before incarceration. The time spent in holding cells is an important opportunity to provide risk reduction counselling and intervention to reduce the incidence of HIV. PMID- 12574044 TI - Serum thyroid stimulating hormone in assessment of severity of tissue hypothyroidism in patients with overt primary thyroid failure: cross sectional survey. PMID- 12574045 TI - Cardiac troponin T in the severity assessment of patients with pulmonary embolism: cohort study. PMID- 12574047 TI - Validation of the Fresno test of competence in evidence based medicine. AB - OBJECTIVE: To describe the development and validation of a test of knowledge and skills in evidence based medicine. DESIGN: Cross sectional study. SETTING: Family practice residency programme in California; a list server for those who teach evidence based medicine; and an evidence based medicine seminar series. PARTICIPANTS: Family practice residents and faculty members (n=43); volunteers self identified as experts in evidence based medicine (n=53); family practice teachers (19) beginning a seminar series on evidence based medicine. INTERVENTION: The Fresno test is a performance based measure for use in medical education that assesses a wide range of evidence based medicine skills. Open ended questions are scored with standardised grading rubrics. Calculation skills are assessed by fill in the blank questions. MAIN OUTCOME MEASURES: Inter-rater reliability, internal reliability, item analyses, and construct validity. RESULTS: Inter-rater correlations ranged from 0.76 to 0.98 for individual items. Cronbach's alpha was 0.88. Item difficulties ranged from moderate to difficult, all with positive and strong ability to discriminate between candidates. Experts scored consistently higher than novices. On the 212 point test, the novice mean was 95.6 and the expert mean was 147.5 (P<0.001). On individual items, a higher proportion of experts than novices earned passing scores on 15 of the 17 items. CONCLUSION: The Fresno test is a reliable and valid test for detecting the effect of instruction in evidence based medicine. Its use in other settings requires further exploration. PMID- 12574046 TI - Practice based, longitudinal, qualitative interview study of computerised evidence based guidelines in primary care. AB - OBJECTIVE: To understand the factors influencing the adoption of a computerised clinical decision support system for two chronic diseases in general practice. DESIGN: Practice based, longitudinal, qualitative interview study. SETTING: Five general practices in north east England. PARTICIPANTS: 13 respondents (two practice managers, three nurses, and eight general practitioners) gave a total of 19 semistructured interviews. 40 people in practices included in the randomised controlled trial (34 doctors, three nurses) and interview study (three doctors, one previously interviewed) gave feedback. RESULTS: Negative comments about the decision support system significantly outweighed the positive or neutral comments. Three main areas of concern among clinicians emerged: timing of the guideline trigger, ease of use of the system, and helpfulness of the content. Respondents did not feel that the system fitted well within the general practice context. Experience of "on-demand" information sources, which were generally more positively viewed, informed the comments about the system. Some general practitioners suggested that nurses might find the guideline content more clinically useful and might be more prepared to use a computerised decision support system, but lack of feedback from nurses who had experienced the system limited the ability to assess this. CONCLUSIONS: Significant barriers exist to the use of complex clinical decision support systems for chronic disease by general practitioners. Key issues include the relevance and accuracy of messages and the flexibility to respond to other factors influencing decision making in primary care. PMID- 12574048 TI - Making decisions about hormone replacement therapy. PMID- 12574049 TI - Misdiagnosis of epilepsy in patients prescribed anticonvulsant drugs for other reasons. PMID- 12574050 TI - Problem based learning. PMID- 12574051 TI - Prenatal diagnosis requests for Huntington's disease when the father is at risk and does not want to know his genetic status: clinical, legal, and ethical viewpoints. PMID- 12574052 TI - Systematic review of celecoxib for osteoarthritis and rheumatoid arthritis. Problems compromise review's validity. PMID- 12574053 TI - General practitioner screening for excessive alcohol use. Paper enables open debate about a complex intervention. PMID- 12574054 TI - Antiretroviral therapy: new solutions bring new problems. PMID- 12574055 TI - Model for Bangalore helped disseminate information to doctors in India. PMID- 12574056 TI - Depressed patients need more than drugs and psychiatrists. PMID- 12574057 TI - Host should also protect students on electives from HIV. PMID- 12574058 TI - Disposal of remains of fetuses of under 24 weeks' gestation. PMID- 12574059 TI - Surgical oncology. PMID- 12574060 TI - Obstetrics and gynaecology: where are we now? PMID- 12574063 TI - Liquid-liquid immiscibility in membranes. AB - The observation of liquid-liquid immiscibility in cholesterol-phospholipid mixtures in monolayers and bilayers has opened a broad field of research into their physical chemistry. Some mixtures exhibit multiple immiscibilities. This unusual property has led to a thermodynamic model of "condensed complexes." These complexes are the consequence of an exothermic, reversible reaction between cholesterol and phospholipids. In this quantitative model the complexes are sometimes concentrated in a separate liquid phase. The phase separation into a complex-rich phase depends on membrane composition and intensive variables such as temperature. The properties of defined cholesterol-phospholipid mixtures provide a conceptual foundation for the exploration of a number of aspects of the biophysics and biochemistry of animal cell membranes. PMID- 12574064 TI - Computer simulations of enzyme catalysis: methods, progress, and insights. AB - Understanding the action of enzymes on an atomistic level is one of the important aims of modern biophysics. This review describes the state of the art in addressing this challenge by simulating enzymatic reactions. It considers different modeling methods including the empirical valence bond (EVB) and more standard molecular orbital quantum mechanics/molecular mechanics (QM/MM) methods. The importance of proper configurational averaging of QM/MM energies is emphasized, pointing out that at present such averages are performed most effectively by the EVB method. It is clarified that all properly conducted simulation studies have identified electrostatic preorganization effects as the source of enzyme catalysis. It is argued that the ability to simulate enzymatic reactions also provides the chance to examine the importance of nonelectrostatic contributions and the validity of the corresponding proposals. In fact, simulation studies have indicated that prominent proposals such as desolvation, steric strain, near attack conformation, entropy traps, and coherent dynamics do not account for a major part of the catalytic power of enzymes. Finally, it is pointed out that although some of the issues are likely to remain controversial for some time, computer modeling approaches can provide a powerful tool for understanding enzyme catalysis. PMID- 12574065 TI - Proteome analysis by mass spectrometry. AB - The coupling of high-performance mass spectrometry instrumentation with highly efficient chromatographic and electrophoretic separations has enabled rapid qualitative and quantitative analysis of thousands of proteins from minute samples of biological materials. Here, we review recent progress in the development and application of mass spectrometry-based techniques for the qualitative and quantitative analysis of global proteome samples derived from whole cells, tissues, or organisms. Techniques such as multidimensional peptide and protein separations coupled with mass spectrometry, accurate mass measurement of peptides from global proteome digests, and mass spectrometric characterization of intact proteins hold great promise for characterization of highly complex protein mixtures. Advances in chemical tagging and isotope labeling techniques have enabled quantitative analysis of proteomes, and highly specific isolation strategies have been developed aimed at selective analysis of posttranslationally modified proteins. PMID- 12574066 TI - The structure of mammalian cyclooxygenases. AB - Cyclooxygenases-1 and -2 (COX-1 and COX-2, also known as prostaglandin H2 synthases-1 and -2) catalyze the committed step in prostaglandin synthesis. COX-1 and -2 are of particular interest because they are the major targets of nonsteroidal antiinflammatory drugs (NSAIDs) including aspirin, ibuprofen, and the new COX-2-selective inhibitors. Inhibition of the COXs with NSAIDs acutely reduces inflammation, pain, and fever, and long-term use of these drugs reduces the incidence of fatal thrombotic events, as well as the development of colon cancer and Alzheimer's disease. In this review, we examine how the structures of COXs relate mechanistically to cyclooxygenase and peroxidase catalysis and how alternative fatty acid substrates bind within the COX active site. We further examine how NSAIDs interact with COXs and how differences in the structure of COX 2 result in enhanced selectivity toward COX-2 inhibitors. PMID- 12574067 TI - Optical single transporter recording: transport kinetics in microarrays of membrane patches. AB - Optical single transporter recording (OSTR) is an emerging technique for the fluorescence microscopic measurement of transport kinetics in membrane patches. Membranes are attached to transparent microarrays of cylindrical test compartments (TCs) approximately 0.1-100 mum in diameter and approximately 10-100 mum in depth. Transport across membrane patches that may contain single transporters or transporter populations is recorded by confocal microscopy. By these means transport of proteins through single nuclear pore complexes has been recorded at rates of <1 translocation/s. In addition to the high sensitivity in terms of measurable transport rates OSTR features unprecedented spatial selectivity and parallel processing. This article reviews the conceptual basis of OSTR and its realization. Applications to nuclear transport are summarized. The further development of OSTR is discussed and its extension to a diversity of transporters, including translocases and ATP-binding cassette (ABC) pumps, projected. PMID- 12574069 TI - Molecular recognition and docking algorithms. AB - Molecular docking is an invaluable tool in modern drug discovery. This review focuses on methodological developments relevant to the field of molecular docking. The forces important in molecular recognition are reviewed and followed by a discussion of how different scoring functions account for these forces. More recent applications of computational chemistry tools involve library design and database screening. Last, we summarize several critical methodological issues that must be addressed in future developments. PMID- 12574068 TI - The crystallographic model of rhodopsin and its use in studies of other G protein coupled receptors. AB - G protein-coupled receptors (GPCRs) are integral membrane proteins that respond to environmental signals and initiate signal transduction pathways activating cellular processes. Rhodopsin is a GPCR found in rod cells in retina where it functions as a photopigment. Its molecular structure is known from cryo-electron microscopic and X-ray crystallographic studies, and this has reshaped many structure/function questions important in vision science. In addition, this first GPCR structure has provided a structural template for studies of other GPCRs, including many known drug targets. After presenting an overview of the major structural elements of rhodopsin, recent literature covering the use of the rhodopsin structure in analyzing other GPCRs will be summarized. Use of the rhodopsin structural model to understand the structure and function of other GPCRs provides strong evidence validating the structural model. PMID- 12574070 TI - Surfactant homeostasis is maintained in vivo during keratinocyte growth factor induced rat lung type II cell hyperplasia. AB - Keratinocyte growth factor (KGF) induces transient proliferation of alveolar type II cells (AEII) associated with surfactant alterations. To test the hypothesis that homeostasis of intracellular phospholipid stores is maintained under KGF induced hyperplasia, we (1) collected tissue from adult rat lungs, fixed for light and electron microscopy 3 days after intratracheal instillation of 5 mg recombinant human (rHu) KGF/kg body weight or phosphate-buffered saline (PBS), and from untreated control animals (five animals/group) for design-based stereology of AEII and lamellar body (LB) ultrastructure; and (2) we analyzed uptake and distribution of instilled radiolabeled phospholipids. After rHuKGF, AEII-coverage of alveolar walls (PBS:8.3 +/- 3.0%; rHuKGF:30.6 +/- 4.8%) and number of AEII/ml lung volume (PBS:28.5 +/- 6.5 x 10(6); rHuKGF:48.2 +/- 5.8 x 10(6)) were increased (p < 0.008). Number (PBS:97 +/- 25; rHuKGF:54 +/- 7) and volume (PBS:45.3 +/- 13.8 microm(3); rHuKGF:21.0 +/- 4.7 microm(3)) of LBs per cell were decreased (p < 0.008), but not total amount/ml lung volume (PBS:128 +/- 46. 4 x 10(7) microm(3); rHuKGF:103 +/- 34. 7 x 10(7) microm(3)). This was paralleled by a shift to larger LBs. After rHuKGF, radiolabeled phospholipids accumulated in whole lung tissue relative to lavage fluid (p < 0.01). However, less radiolabel was incorporated per cell (p < 0.01). We conclude that under rHuKGF-induced AEII proliferation intracellular surfactant was decreased per single cell, whereas a constant amount was maintained per unit lung volume. We suggest that surfactant homeostasis is regulated at the level of phospholipid transport processes, for example, secretion and reuptake. PMID- 12574071 TI - Neurokinins modulate hyperventilation-induced bronchoconstriction in canine peripheral airways. AB - This study was designed to test the hypotheses that (1) neurokinin (NK) receptor activity modulates hyperventilation-induced bronchoconstriction (HIB) in canine peripheral airways and (2) NK receptor activity is stimulated via hyperventilation-induced eicosanoid production and release. A bronchoscope was used in anesthetized dogs to record peripheral airway resistance (Rp); to test airway reactivity to NK A (NKA), substance P, and hypertonic saline; and to examine HIB before and after combined treatment with NK-1 (CP 99,994) and NK-2 (SR 48,968) receptor antagonists. Bronchoalveolar lavage fluid cells, prostaglandin D2, and cysteinyl leukotrienes from hyperventilated airways pretreated with either vehicle or NK antagonists were also measured. Pretreatment with NK-1 and NK-2 antagonists significantly attenuated HIB and the response to substance P, virtually abolished the response to NKA, and had little effect on the response to HS. Blockade of NK-1 and NK-2 receptors did not affect either the cell profiles or the mediator concentrations recovered in bronchoalveolar lavage fluid after hyperventilation. We conclude that NKs modulate the development of HIB and appear to do so via hyperventilation-induced eicosanoid production and release. PMID- 12574072 TI - Impact of randomized trial results on acute lung injury ventilator therapy in teaching hospitals. AB - Reducing tidal volumes administered to patients with acute lung injury is the only intervention reported to decrease mortality resulting from this life threatening condition. Whereas many medical advances are slowly brought into practice, clinicians in teaching hospitals are often assumed to be early adopters of new medical advances. Our objective was to examine trends in the ventilatory prescription for 398 patients with acute lung injury treated in three teaching hospitals from 1994 to 2001. There was no change in tidal volumes until mid to late 1998, when volumes started to slowly decline at the rate of 48.0 (95% confidence interval, 21.0 to 74.4) ml/year. In the 2 years after the results were released from a large trial that demonstrated the superiority of 6 ml/kg tidal volume therapy over 12 ml/kg, clinicians prescribed tidal volumes of 651 +/- 128 ml or 10.1 +/- 1.9 ml/kg. Tidal volumes after intubation were minimally reduced over the subsequent 2 days of mechanical ventilation (mean reduction, 33 ml). Hospital category, male sex, and disease onset before May 1999 were associated with higher volumes whereas lung injury severity was inversely associated. We conclude that clinicians practicing at these teaching hospitals have not rapidly adopted low tidal volume ventilation that may reduce mortality. PMID- 12574073 TI - Clinical and biological heterogeneity in children with moderate asthma. AB - To evaluate the relationship between inflammatory markers and severity of asthma in children, the amount of interleukin-8 (IL-8) and granulocyte/macrophage colony stimulating factor (GM-CSF) released by peripheral blood mononuclear cells, exhaled nitric oxide (FE NO) levels, p65 nuclear factor-kappaB subunit, and phosphorylated IkBalpha expression by peripheral blood mononuclear cells were assessed in six control subjects, 12 steroid-naives subjects with intermittent asthma, and 17 children with moderate asthma. To investigate their predictive value, biomarker levels were correlated with the number of exacerbations during a 18-month follow-up period. We found that GM-CSF release was higher in moderate and intermittent asthmatics than in control subjects, whereas IL-8 release was higher in moderate than in intermittent asthmatics and control subjects. FE NO levels were similar among study groups. In moderate asthmatics, IL-8, GM-CSF, and FE NO significantly correlated with the exacerbation numbers. Moreover, p65 and phosphorylated IkBalpha levels were greater in moderate than in intermittent asthmatics and control subjects. According to GM-CSF, IL-8, and FE NO levels, two distinct subgroups of moderate asthmatics (low and high producers) were identified. High producers experienced more exacerbations than low producers. This study shows ongoing inflammation associated with biological and clinical heterogeneity in moderate asthmatics despite regular treatment and proposes that large prospective studies confirm the importance of biomarkers to assess inflammation and asthma control in children with asthma. PMID- 12574074 TI - Acute monocytic leukemia presenting as acute respiratory failure. AB - Acute respiratory failure revealing acute monocytic leukemia is rare. We report 20 patients admitted to the intensive care unit (ICU) with three remarkable features: (1) rapidly progressive respiratory distress revealing acute leukemia, (2) monocytic leukemia, and (3) respiratory status deterioration after chemotherapy initiation. The median age was 50 years (17-72 years), and respiratory symptoms started 2 days (0-15 days) before ICU admission. The median leukocyte count was 98,250/mm3 (800-529,000), with circulating monocytic cells in all of the patients but one. Bone marrow examination was diagnostic of monocytic leukemia in all patients. At presentation, respiratory rate was 33 (18-50) per minute, and PaO2 on room air was 44.5 mm Hg (30-60). Chest radiographs revealed unilateral alveolar infiltrates (n = 1), bilateral alveolar infiltrates with (n = 3) or without (n = 11) pleural effusion, or diffuse interstitial infiltrates (n = 5). Alveolar hemorrhage was the main bronchoalveolar lavage finding, with monocytic cells retrieved from four patients. Respiratory function deteriorated after cancer chemotherapy initiation in all patients. Of the 15 patients who required mechanical ventilation, 10 died. Leukemic pulmonary infiltration as the first manifestation of acute monocytic leukemia should be recognized, and intensive management should be provided in anticipation of the respiratory function deterioration seen consistently after chemotherapy initiation. PMID- 12574075 TI - Biologically active intercellular adhesion molecule-1 is shed as dimers by a regulated mechanism in the inflamed pleural space. AB - Intercellular adhesion molecule-1 (ICAM-1) is an adhesion molecule that plays a crucial role in cell-cell interactions involved in the recruitment of cells and immune responses. Under some circumstances, ICAM-1 is found as a soluble protein that has the potential to influence the nature of immunoinflammatory responses. By examining cells and fluid from the pleural compartment of patients with cancer, tuberculosis, and congestive heart failure, the cellular source, conformation, control, and biological activity of soluble ICAM-1 (sICAM-1) were investigated. The results suggest that dimeric sICAM-1 was released locally in the pleural compartment of tuberculous and malignant effusions. sICAM-1 was shed from preexpressed surface ICAM-1 rather than produced de novo, and both CD45 positive leukocytes and cytokeratin-positive epithelial and mesothelial cells expressed ICAM-1, suggesting multiple cellular sources for sICAM-1. The expression of sICAM-1 was regulated because pleural macrophages caused release of sICAM-1 via a tumor necrosis factor-alpha-dependent mechanism. The functional significance of sICAM-1 was demonstrated by showing that pleural sICAM-1 interfered with conjugate formation between LAK cells and K562 cells, suggesting that pleural sICAM-1 plays an immunosuppressive role by inhibiting adhesion of cytotoxic lymphocytes and tumor cells. Thus, sICAM-1 is shed from the surface of cells in a regulated manner and has the potential to influence the immune response in the pleural space. PMID- 12574076 TI - Cost-effectiveness analysis of augmentation therapy for severe alpha1-antitrypsin deficiency. AB - A Markov-based decision model was created to assess the cost-effectiveness of augmentation therapy (Aug) for severe alpha1-antitrypsin deficiency, comparing strategies of: (1) no Aug, (2) Aug for life, and (3) Aug until FEV1 is below 35% predicted. A hypothetical cohort of 46-year-old patients with FEV1 49% predicted was followed over time using Monte Carlo simulation across five possible health states: (1) FEV1 50 to 79% predicted, (2) FEV1 35 to 49% predicted, (3) FEV1 below 35% predicted, (4) status-post-lung transplantation, and (5) dead. Treatment for life yielded 7.19 quality-adjusted life-years (QALYs) and cost 895,243 dollars. Treating until FEV1 is below 35% predicted cost 511,930 dollars and produced 6.64 QALYs. "No Aug" cost 92,091 dollars with 4.62 QALYs. The incremental cost-effectiveness ratio was 207,841 dollars/QALY for Aug until FEV1 is below 35% predicted and 312,511 dollars/QALY for the "Aug for life" strategy. In all sensitivity analyses, the incremental cost-effectiveness ratio for Aug for life exceeded 100,000 dollars. The cost of Aug needed to be reduced from 54,765 dollars to 4,900 dollars for the "Aug for life" strategy to be considered cost effective. We conclude that, compared with other conventionally used health interventions, Aug is relatively less cost-effective. These results should encourage the development of more clinically and cost-effective therapies for alpha1-antitrypsin deficiency. PMID- 12574077 TI - High plasma osteopontin level and its relationship with interleukin-12-mediated type 1 T helper cell response in tuberculosis. AB - Osteopontin (OPN, also known as Eta-1), a noncollagenous matrix protein produced by macrophages and T lymphocytes, is expressed in granulomatous lesions caused by Mycobacterium tuberculosis infection. In the present study, we compared plasma concentrations of OPN in patients with active pulmonary tuberculosis with those of healthy control subjects and patients with sarcoidosis, another disease associated with granuloma formation. Plasma OPN levels were significantly higher in patients with tuberculosis (n = 48) than in control subjects (n = 34) and patients with sarcoidosis (n = 20). OPN levels correlated well with severity of pulmonary tuberculosis, as indicated by the size of lung lesions on chest X-ray films. Furthermore, chemotherapy resulted in a significant fall in plasma OPN levels. In patients with tuberculosis, plasma OPN concentrations correlated significantly with those of interleukin (IL)-12. In vitro experiments showed that OPN production by peripheral blood mononuclear cells infected with Mycobacterium bovis bacillus Calmette-Guerin preceded the synthesis of IL-12 and interferon gamma and that the neutralizing anti-OPN monoclonal antibody significantly reduced the production of IL-12 and interferon-gamma. Our results suggest that OPN may be involved in the pathologic process associated with active pulmonary tuberculosis by inducing IL-12-mediated type 1 T helper cell responses. PMID- 12574078 TI - Dexamethasone upregulates 11beta-hydroxysteroid dehydrogenase type 2 in BEAS-2B cells. AB - The actions of natural and synthetic glucocorticoids are in part determined by 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2). We examined whether carbenoxolone, a potent inhibitor of 11beta-HSD, would potentiate the inhibitory action of dexamethasone on interleukin-8 release from BEAS-2B cells, and whether prolonged treatment with dexamethasone at therapeutic doses would upregulate 11beta-HSD2 in the cells. We found that carbenoxolone increased the potency of dexamethasone almost 10-fold. Reverse transcription-polymerase chain reaction and Western blot revealed that BEAS-2B cells expressed 11beta-HSD2, but not 11beta HSD1. An enzyme activity assay of the cell homogenate demonstrated only NAD+ dependent dehydrogenase activity. The Km value for cortisol in intact BEAS-2B cells was estimated to be 42 nM. When the cells were incubated with dexamethasone for up to 72 hours at increasing concentrations (10(-9) to 10(-5) M), there were considerable increases in mRNA and protein levels of 11beta-HSD2. Prolonged treatment with dexamethasone also increased the enzyme activity of 11beta-HSD in the cells in a dose- and time-dependent manner, with complete inhibition by RU38486. These results suggest that bronchial epithelial cells possess an autoregulatory system for glucocorticoids in the control of their own bioactive levels by inducing the expression of 11beta-HSD2, and that 11beta-HSD2 in the bronchial epithelium may play a role in the local regulation of inhaled glucocorticoid actions. PMID- 12574079 TI - False claims of blood pressure-independent protection by blockade of the renin angiotensin aldosterone system? PMID- 12574080 TI - Loosening the cuff: important new advances in modeling antihypertensive treatment effects in genetic studies of hypertension. PMID- 12574081 TI - Hypertension, platelets, and the endothelium: the "thrombotic paradox" of hypertension (or "Birmingham paradox") revisited. PMID- 12574082 TI - Renoprotection by ACE inhibition or aldosterone blockade is blood pressure dependent. AB - Renin-angiotensin-aldosterone system blockade has been shown to protect against renal damage in salt-supplemented, stroke-prone spontaneously hypertensive rats (SHRsp). Based on intermittent tail-cuff blood pressure (BP) measurements, it has been claimed that such protection is BP-independent and mediated by a blockade of the direct tissue-damaging effects of angiotensin and/or aldosterone. BP radiotelemetry was performed for 8 weeks in approximately 10-week-old male SHRsp who received a standard diet and either tap water (n=10) or 1% NaCl to drink. Saline-drinking SHRsp were either left untreated (n=12), received enalapril (50 mg/L) in drinking fluid (n=9), or had subcutaneous implantation of time-release 200-mg pellets of aldactone (n=10). The average systolic BP (mean+/-SEM) during the final 3 weeks was significantly higher (P<0.05) in untreated saline-drinking (215+/-6 mm Hg) SHRsp but not aldactone-treated (198+/-4 mm Hg) or enalapril treated treated SHRsp (173+/-1 mm Hg), as compared with tap water-drinking SHRsp (197+/-3 mm Hg). Histological renal damage scores at 8 weeks paralleled the BP in all groups, with an excellent correlation (r=0.8, P<0.001, n=41). Moreover, a renal damage score of >5 was only observed in SHRsp whose average systolic BP during the final 3 weeks exceeded 200 mm Hg, indicating a threshold relation with BP. These data show that protection by renin-angiotensin-aldosterone system blockade in this model is BP-dependent and mediated by preventing the severe increases in BP seen in untreated salt-supplemented SHRsp and further underscore the limitations of interpretations based on conventional tail-cuff BP measurements. PMID- 12574083 TI - Antihypertensive treatments obscure familial contributions to blood pressure variation. AB - The linkage and association between inherent blood pressure and underlying genotype is potentially confounded by antihypertensive treatment. We estimated blood pressure variance components (genetic, shared environmental, individual specific) in 767 adult volunteer families by using a variety of approaches to adjusting blood pressure of the 244 subjects (8.2%) receiving antihypertensive medications. The additive genetic component of variance for systolic pressure was 73.9 mm Hg(2) (SE, 8.8) when measured pressures (adjusted for age by gender within each generation) were used but fell to 61.4 mm Hg(2) (SE, 8.0) when treated subjects were excluded. When the relevant 95th percentile values were substituted for treated systolic pressures, the additive genetic component was 81.9 mm Hg(2) (SE, 9.5), but individual adjustments in systolic pressure ranged from -53.5 mm Hg to +64.5 mm Hg (mean, +17.2 mm Hg). Instead, when 10 mm Hg was added to treated systolic pressure, the additive genetic component rose to 86.6 mm Hg(2) (SE, 10.1). Similar changes were seen in the shared environment component of variance for systolic pressure and for the combined genetic and shared environmental (ie, familial) components of diastolic pressure. There was little change in the individual-specific variance component across any of the methods. Therefore, treated subjects contribute important information to the familial components of blood pressure variance. This information is lost if treated subjects are excluded and obscured by treatment effects if unadjusted measured pressures are used. Adding back an appropriate increment of pressure restores familial components, more closely reflects the pretreatment values, and should increase the power of genomic linkage and linkage disequilibrium analyses. PMID- 12574084 TI - Effects of severe hypertension on endothelial and platelet microparticles. AB - The molecular mechanisms by which extreme blood pressure elevation leads to vascular injury are not defined. To explore the hypothesis that activation of endothelium and platelets as manifested by increased concentrations of circulating endothelial microparticles and platelet microparticles could play a role in this target organ injury, we conducted a cross-sectional study of these markers in 3 groups: (1) untreated patients referred specifically for treatment of severe uncontrolled hypertension; (2) untreated patients with established mild hypertension; and (3) normotensive volunteer subjects. By ANOVA, endothelial (P=0.002) and platelet (P=0.01) microparticles were greatest in the severely hypertensive group. There was a significant correlation between both of these markers and blood pressure, even in the setting of multiple risk factors. Our results suggest that these markers may be useful and specific for pressure induced endothelial and platelet activation in hypertension. Furthermore, because of the combined effects of endothelial and platelet microparticles on coagulation, leukocytes, and endothelium, it is possible that they may play a pathogenic role in mediating target organ injury in severe hypertension. PMID- 12574085 TI - Atrial fibrillation in hypertension: predictors and outcome. AB - Incidence, determinants, and outcome of atrial fibrillation in hypertensive subjects are incompletely known. We followed for up to 16 years 2482 initially untreated subjects with essential hypertension. At entry, all subjects were in sinus rhythm. Subjects with valvular heart disease, coronary artery disease, preexcitation syndrome, thyroid disorders, or lung disease were excluded. During follow-up, a first episode of atrial fibrillation occurred in 61 subjects at a rate of 0.46 per 100 person-years. At entry, subjects with future atrial fibrillation differed (all P<0.05) from those without by age (59 versus 51 years), office, and 24-hour systolic blood pressure (165 and 144 versus 157 and 137 mm Hg, respectively), left ventricular mass (58 versus 49 g/height[m](2.7)), and left atrial diameter (3.89 versus 3.56 cm). Age and left ventricular mass (both P<0.001) were the sole independent predictors of atrial fibrillation. For every 1 standard deviation increase in left ventricular mass, the risk of atrial fibrillation was increased 1.20 times (95% CI, 1.07 to 1.34). Atrial fibrillation became chronic in 33% of subjects. Age, left ventricular mass, and left atrial diameter (all P<0.01) were independent predictors of chronic atrial fibrillation. Ischemic stroke occurred at a rate of 2.7% and 4.6% per year, respectively, among subjects with paroxysmal and chronic atrial fibrillation. These data indicate that in hypertensive subjects with sinus rhythm and no other major predisposing conditions, risk of atrial fibrillation increases with age and left ventricular mass. Increased left atrial size predisposes to chronicization of atrial fibrillation. PMID- 12574086 TI - Coronary vasodilator capacity and hypertension-induced increase in left ventricular mass. AB - An increase in left ventricular mass represents a compensatory response of hypertensive heart to augmented loading conditions. The concept of inappropriate mass has been proposed to define an increase in left ventricular mass higher than needed to compensate for increased workload. To assess whether inappropriate left ventricular mass is associated with more severe impairment of coronary vasodilator capacity, 64 untreated middle-aged hypertensive patients without significant coronary artery stenosis and 14 normotensive volunteers comparable for age and gender were studied by transthoracic and transesophageal echocardiography to evaluate left ventricular mass, geometry, and coronary flow velocity response to adenosine. Thirty-three patients had appropriate and 31 had inappropriate increase in left ventricular mass, whereas all normotensive control subjects had appropriate left ventricular mass. Compared with control subjects, minimum coronary resistance (0.87+/-0.18 mm Hg per second/centimeter) was increased in both hypertensive subgroups, more in those with inappropriate left ventricular mass (1.34+/-0.23 versus 1.19+/-0.23 mm Hg per second/centimeter, P<0.01), who also exhibited lower afterload-corrected midwall shortening and ratio of peak early and peak late velocities of transmitral flow profile. In hypertensive patients, minimum coronary resistance was related positively to absolute and relative left ventricular wall thickness (r=+0.33 and +0.35, both P<0.01) and negatively to midwall shortening and ratio of peak early and peak late velocities of transmitral flow (r=-0.32 and -0.31, both P<0.02). Thus, in the hypertensive heart, a deviation of left ventricular mass from values compensatory for increased cardiac workload is associated with lower coronary vasodilator capacity, depressed left ventricular wall mechanics, and abnormal left ventricular diastolic filling pattern. PMID- 12574087 TI - Myocardial ultrasonic backscatter in hypertension: relation to aldosterone and endothelin. AB - A disproportionate accumulation of fibrillar collagen is a characteristic feature of hypertensive heart disease, but the extent of myocardial fibrosis may differ in different models of hypertension. In experimental studies, aldosterone and endothelins emerge as important determinants of myocardial fibrosis. Changes in myocardial extracellular matrix and collagen deposition can be estimated noninvasively by analysis of the ultrasonic backscatter signal, which arises from tissue heterogeneity within the myocardium and describes myocardial texture. This study was designed to investigate the relations between myocardial integrated backscatter and circulating aldosterone and immunoreactive endothelin in human hypertension. The study population consisted of 56 subjects: 14 healthy normotensive volunteers and 42 hypertensive patients (14 with primary aldosteronism, 7 with renovascular hypertension, and 21 with essential hypertension). The patients with essential and secondary hypertension were matched for age, gender, body mass index, and blood pressure. Myocardial integrated backscatter at diastole was 19.8+/-2.0 and 20.8+/-2.9 decibels in normotensive control subjects and patients with essential hypertension and significantly higher in patients with primary aldosteronism (27.4+/-3.8 decibels, P<0.01) and renovascular hypertension (26.8+/-4.8 decibels, P<0.01). In the population as a whole, as well as in the hypertensive subpopulation, myocardial integrated backscatter was directly related to plasma aldosterone (r=0.73 and 0.71, P<0.01 for both) and immunoreactive endothelin (r=0.60 and 0.56, P<0.01 for both). The data of this study suggest that in human hypertension, circulating aldosterone and immunoreactive endothelin may induce alterations in left ventricular myocardial texture, possibly related to increased myocardial collagen content. PMID- 12574088 TI - Morning blood pressure peak, QT intervals, and sympathetic activity in hypertensive patients. AB - We investigated the relation between morning blood pressure (BP) variations, sympathetic activity, and QT intervals in 156 never-treated subjects with essential hypertension and different patterns of morning BP increase. The morning BP peak (MP) was defined as a rise in systolic BP >or=50 mm Hg and/or diastolic BP >or=22 mm Hg during early morning (6:00 to 10:00 AM) compared with mean BP during the night. Clinical characteristics of patients with morning BP peak (MP+, n= 69, morning systolic BP=+54+/-4, diastolic BP=+32+/-5 mm Hg) did not differ from patients without BP peak (MP-, n= 87, morning systolic BP=+24+/-5, diastolic BP=+19+/-3 mm Hg). The daytime (10:00 AM to 10:00 PM) and the nighttime (10:00 PM to 6:00 AM) BP profile did not differ between the two groups. During daytime and nighttime ECG monitoring, the corrected QT (QTc) interval, and QTc dispersion did not differ significantly between the two groups, whereas during the morning period the QT values were significantly broader in the MP+ group compared with the MP- group (P3 times the increase in the vehicle rats. Heart rate also increased significantly in the L rats, from 391+/-4 to 426+/-8 bpm, and that increase was prevented completely in the LB rats. However, although the increase in MAP in the LB rats was significantly less than in the L rats, the hypertension was not prevented completely. The explanation for that partial inhibition is not clear, but the overall effectiveness of adrenergic receptor blockade to attenuate the potentiated hypertensive and tachycardic responses to glucose infusion in the L-NAME-treated rats versus the normal rats suggests that nitric oxide may help protect against hypertension during glucose infusion through suppression of sympathetic activity. PMID- 12574095 TI - Guanylyl cyclase receptors mediate cardiopulmonary vagal reflex actions of ANP. AB - Atrial natriuretic peptide (ANP) potentiates vagal cardiopulmonary reflexes due to chemosensory (Bezold-Jarisch [B-J] reflex) or mechanosensory (ramp baroreflex) activation. The ANP receptor mediating these actions is unknown. We examined the role of particulate guanylyl-cyclase (pGC) receptors in ANP-induced enhancement of cardiopulmonary vagal reflexes. Cardiopulmonary baroreceptor reflex function was assessed by bradycardic responses to ramp blood pressure rises after rapid intravenous methoxamine (100 micro g/kg bolus dose). The B-J reflex was evoked by 3 intravenous doses of serotonin (1 to 10 micro g/kg). In conscious, chronically instrumented rats (n=9), these tests were performed on each animal during randomized infusions of rat ANP (150 ng/kg per minute IV), saline (270 micro L/h IV), the pGC receptor antagonist HS-142-1 (3 mg/kg IV), or combined HS-142-1+ANP treatment. HS-142-1 alone attenuated normal B-J reflex (by 33+/-8%, P<0.05) but not ramp baroreflex responses. As we showed previously, ANP enhanced baroreflex and B-J reflex bradycardia (by approximately 140% and approximately 30%, respectively, P<0.05), compared with saline infusion. These ANP effects were completely blocked by HS-142-1, demonstrating that the cardiopulmonary vagal reflex actions of ANP occurred through pGC natriuretic peptide receptors. Additionally, we have provided evidence for the first time that pGC natriuretic peptide receptors are essential for the full expression of the B-J reflex but not for that of cardiopulmonary vagal baroreflexes. This tonic interaction between pGC natriuretic peptide receptors and cardiopulmonary chemosensitive receptors may be important during pathophysiological activation of B-J reflex, such as with myocardial infarction. PMID- 12574096 TI - Lipid peroxidation is not increased in patients with untreated mild-to-moderate hypertension. AB - In contrast with the huge amount of experimental data available, only few and somewhat unconvincing clinical studies support the hypothesis that oxidative stress is involved in the early stages of essential hypertension in humans. Isoprostanes are chemically stable lipid peroxidation products of arachidonic acid, the quantification of which provides a novel approach to the assessment of oxidative stress in vivo. The main objective of this study was to quantify the urinary levels of 15-F(2t)-IsoP in the early stages of essential hypertension, using gas chromatography/mass spectrometry, by comparing 30 patients with never treated mild-to-moderate hypertension with 30 gender- and age-paired healthy controls. Urinary 15-F(2t)-IsoP levels were not significantly different in hypertensive patients (69+/-36 pmol/mmol creatinine) compared with controls (75+/ 34 pmol/mmol creatinine, 95% confidence intervals on differences: -23 to 13). No significant correlation was found between basal urinary 15-F(2t)-IsoP levels and age, low-density lipoprotein cholesterol, glucose, clinical pulse pressure, carotid intima-media thickness, left ventricular mass index, or aortic pulse wave velocity. In conclusion, this study shows that lipid peroxidation is not increased in never-treated mild-to-moderate hypertension. This suggests that oxidative stress is not implicated in the pathogenesis of human essential hypertension, at least in the early stages. PMID- 12574097 TI - Radical-trapping activity, blood pressure, and carotid enlargement in women. AB - The aim of this study was to evaluate the influence of traditional and nontraditional (oxidation markers) cardiovascular risk factors on the degree of adaptive response of the carotid wall to atherosclerotic disease, a process known as arterial enlargement. Five thousand sixty-two clinically healthy, middle-aged women living in the area of Naples participated in the "Progetto Atena" study; 310 of these women (potentially at higher atherosclerotic risk) underwent a high resolution ultrasound scan of the carotid arteries. In addition to routine biochemical tests, these women had the determination of serum IgG antibody titer against oxidized LDL and measurement of thiobarbituric acid reactive substances and total radical-trapping activity potential of plasma. Age, systolic blood pressure, body mass index, and radical-trapping activity were all positively correlated with external and internal common carotid diameters, whereas triglycerides (positively) and HDL cholesterol (inversely) were related only to external diameter. After controlling for traditional cardiovascular risk factors, associations still persisted for age, systolic blood pressure, and plasma radical trapping activity with external carotid diameters. However, in the quartile of women with highest total cholesterol (>7.38 mmol/L), the slope of the regression line between systolic blood pressure and external diameter was significantly flatter than in the three other quartiles (test for difference, P=0.014). Outward carotid enlargement is related to traditional and nontraditional risk factors and comes even before plaque development. Women with poor resistance to oxidative stress potentially have a difficulty to remodel their arteries in response to atherosclerotic stimuli. PMID- 12574098 TI - Effects of isosorbide mononitrate and AII inhibition on pulse wave reflection in hypertension. AB - The aortic pulse wave contour in isolated systolic hypertension often shows a prominent reflection peak, which combines with the incident wave arising from cardiac ejection so as to widen pulse pressure. We investigated the effects of an extended-release nitrate preparation and of 2 angiotensin II (AII) inhibitors (an AII receptor antagonist and an ACE inhibitor) on the aortic pulse wave contour and systemic blood pressure in hypertensive subjects with high augmentation index caused by exaggerated pulse wave reflection. Two double-blind, randomized, placebo-controlled crossover studies were carried out in a total of 16 elderly patients with systolic hypertension resistant to conventional antihypertensive therapy. In 1 study, pharmacodynamic responses to single doses of placebo, isosorbide mononitrate, eprosartan, and captopril were determined; in the other, single-dose isosorbide mononitrate and placebo were compared in subjects treated with AII inhibitors at baseline. Blood pressure was measured by sphygmomanometry and pulse wave components by applanation tonometry at the radial artery. All 3 agents were shown to decrease brachial systolic blood pressure, aortic systolic blood pressure, and aortic pulse pressure. Qualitative effects on the aortic pulse wave contour differed: augmentation index was not significantly altered by either captopril or eprosartan but was decreased (P<0.0001) by approximately 50% of the placebo value with isosorbide mononitrate in both study groups. We propose that isosorbide mononitrate corrected the magnified wave reflection in systolic hypertension of these elderly patients by an effect that was distinct from that exercised by either acute or chronic AII inhibition. PMID- 12574099 TI - Adrenomedullin overexpression to inhibit cuff-induced arterial intimal formation. AB - Adrenomedullin (AM) inhibits vascular smooth muscle cell proliferation stimulated by fetal calf serum and platelet-derived growth factor in vitro. In this study, an adenovirus expressing AM (AxCAAM) was created to examine the in vivo action of AM. Femoral arteries of Wistar rats were wrapped with a silicone cuff and treated with adenovirus expressing Escherichia coli beta-galactosidase (AxCALacZ) or AxCAAM. Immunoreactivity for endothelial nitric oxide synthase (eNOS) was reduced in the endothelium of cuff-injured arteries and was associated with increased local DNA synthesis. Consequently, the intimal formation measured by the intimal to-medial ratio was significantly increased at 14 and 28 days after the cuff placement. AxCAAM-infected arteries increased the expression of eNOS in the endothelium and inducible NOS in the media and the adventitia. AxCAAM significantly decreased the intimal-to-medial ratio by 40% at 14 days and 51% at 28 days, whereas AxCALacZ showed no changes compared with cuff-injured control arteries. AM overexpression effectively limits intimal hyperplasia by reducing cell proliferation through a nitric oxide-dependent pathway of eNOS. Our findings suggest the possibility of a therapeutic use of the AM gene for the prevention of vascular proliferative disorders. PMID- 12574100 TI - Haplotype analysis of the human renin gene and essential hypertension. AB - The human renin gene is an attractive candidate for involvement in the underlying cause of essential hypertension (EH). Despite extensive examination, the relation between the renin gene and hypertension remains unclear. The aims of the present study were to discover new genetic markers of EH and to investigate the relations between polymorphisms of the renin gene and EH in the Japanese. Using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method, we isolated 3 novel variants of the renin gene; a single nucleotide polymorphism (SNP) in intron 4 (T+17int4G), a variable number of tandem repeats (VNTR) polymorphism in intron 7, and a missense mutation in exon 9 (G1051A). We performed an association study with these polymorphisms in 212 patients with EH and 209 age-matched normotensive (NT) subjects. The frequency of genotypes VNTR and T+17int4G did not differ significantly between the 2 groups, whereas the overall distribution of G1051A was significantly different between EH and NT. Haplotype analysis revealed that the overall distribution of haplotypes differed significantly between the EH and NT groups. PRA levels in patients with EH with the G/G genotype were significantly higher than in subjects with EH with G/A and A/A genotypes. These data suggest that the missense mutation in exon 9 may affect the enzymatic function of renin and consequently may be involved in the etiology of hypertension. PMID- 12574101 TI - Mice lacking endothelial ACE: normal blood pressure with elevated angiotensin II. AB - Recently, the concept of local renin-angiotensin systems (RAS) capable of generating angiotensin II apart from the circulation has received considerable attention. To investigate this, we generated ACE 1/3 mice in which one allele of ACE is null and the second allele was engineered to express ACE on the surface of hepatocytes. ACE 1/3 mice express no endothelial ACE and lack ACE within the lungs. Their kidneys contain <7.8% the enzyme levels present in control mice. Plasma conversion of angiotensin I to angiotensin II was 43.3% normal. The baseline blood pressure and renal function of the ACE 1/3 mice were normal, probably as a function of a marked increase of both plasma angiotensin I and angiotensin II. When exposed to 2 weeks of a salt-free diet (a stress diet stimulating the RAS), blood pressure in ACE 1/3 mice decreased to 92.3+/-2.0 mm Hg, a level significantly lower than that of wild-type control mice. The ACE 1/3 mice demonstrate the plasticity of the RAS and show that significant compensation is required to maintain normal, basal blood pressure in a mouse with an impaired local vascular and renal RAS. PMID- 12574102 TI - Angiotensinogen gene knockout delays and attenuates cold-induced hypertension. AB - The aim of the present study was to assess our hypothesis that the renin angiotensin system (RAS) is responsible for cold-induced hypertension and cardiac hypertrophy. Two groups of wild-type (WT) mice and 2 groups of angiotensinogen gene knockout (Agt-KO) mice (6 per group) were used. After blood pressures (BP) of the four groups were measured 3 times at room temperature (25 degrees C), 1 WT and 1 Agt-KO group were exposed to cold (5 degrees C). The remaining groups were kept at 25 degrees C. BP of the cold-exposed WT group increased significantly in 1 week of cold exposure and rose gradually to 168+/-7 mm Hg by week 5, whereas the BP of the Agt-KO group did not increase until week 3. The cold-induced increase in BP (DeltaBP) was decreased significantly in the Agt-KO mice (19+/-3 mm Hg) compared with that of the WT mice (61+/-5 mm Hg) by 5 weeks of exposure to cold. Both WT and Agt-KO groups had cardiac hypertrophy in cold to the same extent. Agt-KO caused a significant increase in nitric oxide (NO) production. Thus, the RAS may inhibit NO formation. Chronic cold exposure decreased NO production, which may be mediated partially by activation of the RAS. These results strongly support that the RAS plays a critical role in the development of cold-induced hypertension but not cardiac hypertrophy. Moreover, the role of the RAS in cold-induced hypertension may be mediated in part by its inhibition on NO production. The findings also reveal the possible relation between the RAS and NO in cardiovascular regulation. PMID- 12574103 TI - Prenatal dexamethasone programs hypertension and renal injury in the rat. AB - Dexamethasone is frequently administered to the developing fetus to accelerate pulmonary development. The purpose of the present study was to determine if prenatal dexamethasone programmed a progressive increase in blood pressure and renal injury in rats. Pregnant rats were given either vehicle or 2 daily intraperitoneal injections of dexamethasone (0.2 mg/kg body weight) on gestational days 11 and 12, 13 and 14, 15 and 16, 17 and 18, or 19 and 20. Offspring of rats administered dexamethasone on days 15 and 16 gestation had a 20% reduction in glomerular number compared with control at 6 to 9 months of age (22 527+/-509 versus 28 050+/-561, P<0.05), which was comparable to the percent reduction in glomeruli measured at 3 weeks of age. Six- to 9-month old rats receiving prenatal dexamethasone on days 17 and 18 of gestation had a 17% reduction in glomeruli (23 380+/-587) compared with control rats (P<0.05). Male rats that received prenatal dexamethasone on days 15 and 16, 17 and 18, and 13 and 14 of gestation had elevated blood pressures at 6 months of age; the latter group did not have a reduction in glomerular number. Adult rats given dexamethasone on days 15 and 16 of gestation had more glomeruli with glomerulosclerosis than control rats. This study shows that prenatal dexamethasone in rats results in a reduction in glomerular number, glomerulosclerosis, and hypertension when administered at specific points during gestation. Hypertension was observed in animals that had a reduction in glomeruli as well as in a group that did not have a reduction in glomerular number, suggesting that a reduction in glomerular number is not the sole cause for the development of hypertension. PMID- 12574104 TI - Nephron number, renal function, and arterial pressure in aged GDNF heterozygous mice. AB - The loss of one allele for glial cell line-derived neurotrophic factor (GDNF) results in approximately 30% fewer but normal sized glomeruli in young mice. Low nephron number, inherited or acquired, has been linked to increased risk of development of hypertension and renal failure. This study examines whether GDNF heterozygous mice, with an inherent reduction in nephron number, demonstrate a deterioration in renal structure and function and rise in arterial pressure in later life. Fourteen-month-old male GDNF heterozygous (n=7) and wild-type (n=6) mice were anesthetized and prepared for measurement of mean arterial pressure, glomerular filtration rate (GFR), and renal blood flow. After measurement of renal function, kidneys were fixed for stereological determination of total glomerular number and mean glomerular volume. Mean arterial pressure was, on average, 18 mm Hg higher in GDNF heterozygous (98+/-4 mm Hg) than wild-type mice (80+/-2 mm Hg; P<0.01). However, GFR (0.656+/-0.054 versus 0.688+/-0.076 mL/min per g kidney wt) and renal blood flow (5.29+/-0.42 versus 4.70+/-0.34 mL/min per g kidney wt) were not different between groups. Fourteen-month-old GDNF heterozygous mice had approximately 30% fewer glomeruli than wild-type mice (9206+/-934 versus 13440+/-1275; P<0.01) and significantly larger glomeruli (4.51+/-0.39 versus 3.72+/-0.63x10(-4)mm(3); P<0.01). Thus, aged GDNF heterozygous mice maintained a normal GFR and renal blood flow despite reduced nephron numbers. The elevated arterial pressure, glomerular hypertrophy, and hyperfiltration demonstrated in the GDNF heterozygous mice at this age may indicate a compensatory mechanism whereby GFR is maintained in the presence of a reduced nephron endowment. PMID- 12574105 TI - Antioxidant-rich diet relieves hypertension and reduces renal immune infiltration in spontaneously hypertensive rats. AB - Previous studies have demonstrated that oxidative stress contributes to hypertension and treatments with either antioxidant or immunosuppressive/anti inflammatory agents improve hypertension in spontaneously hypertensive rats (SHR). The present study was performed to determine if the antihypertensive effects of an antioxidant-rich diet are associated with reduction in the renal immune infiltration. Rats were divided into experimental groups (n=5 each) that were followed 7 months after birth, during which they were fed either a regular or antioxidant-enriched (test) diet as follows: SHR-R group=regular diet; SHR-T group=test diet throughout the experiment; SHR-S group=test diet for 4 months switched to regular diet thereafter; WKY group=control rats given regular diet. The SHR-T rats showed a significant reduction in systolic blood pressure (mm Hg): SHR-T=179.6+/-12.9 versus SHR-R=207.5+/-9.6 (P<0.001) and plasma hydrogen peroxide concentration (SHR-T=15+/-4 micro mol/L versus 34+/-9 in SHR-R rats). This was accompanied by significant reductions of renal tissue nitrotyrosine abundance, tubulointerstitial infiltration (cells/mm(2)) of lymphocytes (SHR T=18+/-3 versus SHR-R=30+/-4, P<0.001), macrophages (SHR-T= 17+/-3 versus SHR R=22+/-3), and angiotensin II-positive cells (SHR-T= 17+/-2 versus SHR-R=25+/-5, P<0.01). Results in the SHR-S group were intermediate between the SHR-R and SHR-T groups. The intensity of the infiltration of lymphocytes, macrophages, and angiotensin II-positive cells significantly correlated with systolic blood pressure. Thus, the present study demonstrates that an antioxidant-enriched diet reduces the renal interstitial inflammation and improves hypertension in SHR. These findings point to interrelation between oxidative stress and inflammatory reactivity in the pathogenesis of hypertension. PMID- 12574106 TI - Altered tropomyosin expression in essential hypertension. AB - Abnormal erythrocyte sodium-lithium countertransport is common in a subgroup of patients with essential hypertension and a strong family history of hypertension and cardiovascular disease. We have previously shown that the abnormality in sodium-lithium countertransport is associated with tropomyosin, a cytoskeletal protein required to stabilize actin filament formation. Leukocyte trafficking events, which depend on cytoskeletal reorganization, are also altered in patients with essential hypertension with abnormal sodium-lithium countertransport. The aim of this study was to determine whether there is an abnormality in isoforms of tropomyosin that are common to erythrocytes and leukocytes. Analysis of reticulocyte RNA by reverse transcription (RT) and polymerase chain reaction (PCR) showed expression of TPMN and TPM5b isoforms of tropomyosin. No other isoforms were expressed. These isoforms were also detected in RNA from leukocytes. In patients with essential hypertension with abnormal erythrocyte sodium-lithium countertransport compared with normal control subjects, there was a higher TPMN/TPM5b ratio of protein in erythrocytes (median 3.8 [range 1.8 to 6.6] versus 2.9 [1.9 to 4.0], P<0.001) and of RNA in leukocytes (3.7 [1.7 to 8.2] versus 2.6 [1.2 to 4.3], P<0.01). Furthermore, the protein ratio of TPMN/TPM5b in erythrocytes showed significant correlation with the V(max)/K(m) ratio of sodium lithium countertransport across the patient groups (r=-0.42; P<0.01). Therefore, altered tropomyosin expression may be the underlying abnormality associated with blood cell membrane changes in essential hypertension and implicates the cytoskeleton in the pathogenesis of the disease in a major subgroup of patients. PMID- 12574107 TI - NOS3 genotype-dependent correlation between blood pressure and physical activity. AB - Endothelium-dependent vasorelaxation plays an important role in reduction of blood pressure and is mediated through release of nitric oxide (NO), which is generated by constitutively expressed endothelial nitric oxide synthase (NOS3). Exercise also augments NO release and has been recommended for primary prevention and improvement of hypertension, but individual responses are highly variable. We therefore postulated that genetic polymorphisms of NOS3 might interact with physical activity level to differentially influence blood pressure level. We genotyped 832 healthy Japanese (mean age of 54.4+/-8.6 years, 372 men and 460 women) for a polymorphism of NOS3 in intron 4 (ecNOS4a/b), using the polymerase chain reaction method, and scored their habitual physical activity level by using the rate of energy expenditure per resting metabolic rate through an interview according to a semiquantitative assessment method. Only in the subjects who had the rarer a allele (aa+ba type), systolic blood pressure was found to be inversely correlated with physical activity level (P for linear trend=0.0496, for interaction=0.0071). Eventually, this polymorphism was significantly associated with the prevalence of systolic hypertension only in the subjects who were in the lowest tertile of physical activity level (OR=2.4, 95% CI, 1.1 to 5.6, P for interaction=0.0474). In the present study, we found a significant interaction between the genotype and physical activity level on systolic blood pressure. These results might allow a better understanding of the mechanism to improve hypertension by exercise and to thereby reduce the risk of cardiovascular disease. PMID- 12574108 TI - Beta(2)-adrenergic receptor gene polymorphism, age, and cardiovascular phenotypes. AB - Previous studies suggest that variants of the beta(2)-adrenergic receptor (ADRB2) may differently affect functional responses to adrenergic stimulation, thereby possibly modulating cardiovascular and metabolic phenotypes. We examined the hypothesis that G/R16 and Q/E27 polymorphism of ADRB2, or their haplotypes, may modulate blood pressure, cardiovascular structure, and function or metabolic cardiovascular risk factors in the general population. We examined a random sample of the general population (n=571; age, 35 to 64 years). Neither clinic nor 24-hour ambulatory blood pressure was significantly associated with ADRB2 genotypes in the overall population. Cardiac structure and function were also not influenced by ADRB2 polymorphism. After adjustment for potential confounders, association of the R16 allele with higher systolic blood pressure was observed in the subgroup of younger people (below age of 50 years). Haplotype analysis showed that higher blood pressure values were more specifically associated with the presence of R16-Q27. Younger people carrying the R16-Q27 haplotype also showed a trend toward lower heart rate, higher BMI, lower glycemia, and higher trygliceridemia, which is consistent with the hypothesis of a genetic predisposition to reduced cardiovascular and metabolic response to ADRB2 stimulation. This study does not provide evidence of a major role of ADRB2 gene variability in blood pressure modulation. However, association of ADRB2 polymorphism with cardiovascular and metabolic effects can be observed in younger subjects, before the development of age-related decline of ADRB2-mediated activity. Our study emphasizes the necessity of taking into account (patho) physiological changes related to aging (in this case, decreased efficiency of ADRB2 signaling) when analyzing phenotypic effects of genetic variants. PMID- 12574109 TI - Peroxisome proliferator-activated receptor-gamma coactivator-1 gene locus: associations with hypertension in middle-aged men. AB - Peroxisome proliferator-activated receptor-gamma coactivator-1 (PPARGC1/PGC-1) is a transcriptional coactivator of nuclear hormone receptors implicated in blood pressure regulation. We therefore ascertained whether the PPARGC1 gene locus is associated with hypertension. We studied associations of 3 polymorphisms in PPARGC1 transcripts with hypertension in 683 middle-aged men and 530 middle-aged women of a cross-sectional Austrian population. Hypertension was defined by average values of systolic or diastolic ambulatory blood pressure readings (taken between 7 AM and 10 PM) above 140 and/or 90 and/or use of antihypertensive medication. Among the 3 polymorphic sites, genotype distributions associated with Gly482Ser differed by hypertension status in men (P=0.0038), but not in women. The less common Ser482 allele was associated with a modest, but significant, reduction in the prevalence of hypertension in men. The distribution of 3 loci haplotypes also differed in men with and without hypertension (P=0.015). Despite its moderate effect, but because of its high frequency (approximately 64%), the more common risk allele contributed to hypertension in 35% (95% CI 16% to 54%) of our male population. These results suggest, but do not prove, that PPARGC1 participates in blood pressure control, and sequence substitutions at its gene locus confer an increased risk of hypertension to a substantial proportion of men. PMID- 12574110 TI - Regression of hypertensive myocardial fibrosis by Na(+)/H(+) exchange inhibition. AB - We have recently reported that the inhibition of the Na(+)/H(+) exchanger (NHE) during 1 month in spontaneously hypertensive rats (SHR) is followed by regression of cardiomyocyte hypertrophy but not of myocardial fibrosis. The aim of this study was to evaluate whether a treatment of longer duration could reduce myocardial fibrosis and stiffness. SHR received 3.0 mg/kg per day of the specific NHE-1 inhibitor cariporide; the effect on cardiomyocyte cross-sectional area, myocardial collagen volume fraction, collagen synthesis, and myocardial stiffness (length-tension relation in left papillary muscles) was evaluated at several time points (after 1, 2, or 3 months). A slight decrease of approximately 5 mm Hg in systolic blood pressure was observed after 1 month of treatment with no further changes. After 2 and 3 months of treatment, the size of cardiomyocytes remained within normal values and myocardial fibrosis progressively decreased to normal level. Accordingly, myocardial stiffness and the serum levels of the carboxyterminal propeptide of procollagen type I, a marker of collagen type I synthesis, were normalized after 3 months. Left ventricular weight decreased from 910+/-43 (in untreated SHR) to 781+/-21 mg (treated SHR) after 3 months of treatment. No difference in body weight between treated and untreated SHR was observed after this period of treatment. The present data allow us to conclude that in the SHR the administration of an NHE-1 inhibitor for 2 or 3 months leads to the normalization of collagen type I synthesis, myocardial collagen volume fraction, and stiffness. PMID- 12574111 TI - Wall stiffness suppresses Akt/eNOS and cytoprotection in pulse-perfused endothelium. AB - Increased steady shear stress stimulates nitric oxide synthase (eNOS) in part by Akt-dependent phosphorylation. Arteries in vivo are exposed to pulse perfusion (PP) combining phasic shear with stretch. In compliant vessels, enhancing PP lowers vascular tone by stimulating eNOS; whereas in aged, stiff arteries, flow mediated dilation declines and PP is a prominent risk factor. Here, we tested the hypothesis that reduced wall distensibility alters PP-induced eNOS/Akt mechano signaling. Bovine aortic endothelial cells cultured within distensible tubes were exposed to physiological nonreversing steady or PP (7 dynes/cm(2) mean shear, pulse pressure 0 or 90 mm Hgx2 hours) in a custom servo-system. In compliant tubes, PP doubled Akt phosphorylation above nonpulsatile flow levels, whereas P Akt declined to static levels from PP in stiffer tubes. eNOS phosphorylation (S 1179) similarly increased with PP in compliant tubes but was nearly undetectable with increased PP in stiffer tubes. After PP, brief exposure of cells to ultraviolet irradiation (oxidant stress) and subsequent culture revealed cytoprotection in compliant tubes but diffuse cytotoxicity and cell detachment in stiffer tubes. NOS inhibition by L-NAME converted compliant-tube post-UV behavior to that of stiffer tubes. These data provide novel evidence that wall compliance can directionally mediate endothelial Akt/eNOS phosphorylation and mechano signaling. This may help explain increased vascular risks resulting from artery stiffening. PMID- 12574112 TI - Structure of the herpesvirus major capsid protein. AB - Herpes simplex virus-1 (HSV-1) virions are large, complex enveloped particles containing a proteinaceous tegument layer connected to an icosahedral capsid. The major capsid protein, VP5 (149 kDa), makes up both types of capsomere, pentons and hexons. Limited trypsin digestion of VP5 identified a single stable 65 kDa fragment which represents a proposed protein folding nucleus. We report the 2.9 A crystal structure of this fragment and its modeling into an 8.5 A resolution electron cryomicroscopy map of the HSV-1 capsid. The structure, the first for any capsid protein from Herpesviridae, revealed a novel fold, placing herpesviruses outside any of the structurally linked viral groupings. Alterations in the geometrical arrangements of the VP5 subunits in the capsomeres exposes different residues, resulting in the differential association of the tegument and VP26 with the pentons and hexons, respectively. The rearrangements of VP5 subunits required to form both pentavalent and hexavalent capsomeres result in structures that exhibit very different electrostatic properties. These differences may mediate the binding and release of other structural proteins during capsid maturation. PMID- 12574113 TI - Structural basis for endothelial nitric oxide synthase binding to calmodulin. AB - The enzyme nitric oxide synthase (NOS) is exquisitely regulated in vivo by the Ca(2+) sensor protein calmodulin (CaM) to control production of NO, a key signaling molecule and cytotoxin. The differential activation of NOS isozymes by CaM has remained enigmatic, despite extensive research. Here, the crystallographic structure of Ca(2+)-loaded CaM bound to a 20 residue peptide comprising the endothelial NOS (eNOS) CaM-binding region establishes their individual conformations and intermolecular interactions, and suggests the basis for isozyme-specific differences. The alpha-helical eNOS peptide binds in an antiparallel orientation to CaM through extensive hydrophobic interactions. Unique NOS interactions occur with: (i). the CaM flexible central linker, explaining its importance in NOS activation; and (ii). the CaM C-terminus, explaining the NOS-specific requirement for a bulky, hydrophobic residue at position 144. This binding mode expands mechanisms for CaM-mediated activation, explains eNOS deactivation by Thr495 phosphorylation, and implicates specific hydrophobic residues in the Ca(2+) independence of inducible NOS. PMID- 12574114 TI - Homo- and heterotetrameric architecture of the epithelial Ca2+ channels TRPV5 and TRPV6. AB - The molecular assembly of the epithelial Ca(2+) channels (TRPV5 and TRPV6) was investigated to determine the subunit stoichiometry and composition. Immunoblot analysis of Xenopus laevis oocytes expressing TRPV5 and TRPV6 revealed two specific bands of 75 and 85-100 kDa, corresponding to the core and glycosylated proteins, respectively, for each channel. Subsequently, membranes of these oocytes were sedimented on sucrose gradients. Immuno blotting revealed that TRPV5 and TRPV6 complexes migrate with a mol. wt of 400 kDa, in line with a tetrameric structure. The tetrameric stoichiometry was confirmed in an electrophysiological analysis of HEK293 cells co-expressing concatemeric channels together with a TRPV5 pore mutant that reduced Cd(2+) sensitivity and voltage-dependent gating. Immuno precipitations using membrane fractions from oocytes co-expressing TRPV5 and TRPV6 demonstrated that both channels can form heteromeric complexes. Expression of all possible heterotetrameric TRPV5/6 complexes in HEK293 cells resulted in Ca(2+) channels that varied with respect to Ca(2+)-dependent inactivation, Ba(2+) selectivity and pharmacological block. Thus, Ca(2+) transporting epithelia co-expressing TRPV5 and TRPV6 can generate a pleiotropic set of functional heterotetrameric channels with different Ca(2+) transport kinetics. PMID- 12574115 TI - Doc1 mediates the activity of the anaphase-promoting complex by contributing to substrate recognition. AB - The anaphase-promoting complex (APC) is a multisubunit E3 ubiquitin ligase that targets specific cell cycle-related proteins for degradation, regulating progression from metaphase to anaphase and exit from mitosis. The APC is regulated by binding of the coactivator proteins Cdc20p and Cdh1p, and by phosphorylation. We have developed a purification strategy that allowed us to purify the budding yeast APC to near homogeneity and identify two novel APC associated proteins, Swm1p and Mnd2p. Using an in vitro ubiquitylation system and a native gel binding assay, we have characterized the properties of wild-type and mutant APC. We show that both the D and KEN boxes contribute to substrate recognition and that coactivator is required for substrate binding. APC lacking Apc9p or Doc1p/Apc10 have impaired E3 ligase activities. However, whereas Apc9p is required for structural stability and the incorporation of Cdc27p into the APC complex, Doc1p/Apc10 plays a specific role in substrate recognition by APC coactivator complexes. These results imply that Doc1p/Apc10 may play a role to regulate the binding of specific substrates, similar to that of the coactivators. PMID- 12574116 TI - Mad2 phosphorylation regulates its association with Mad1 and the APC/C. AB - Improper attachment of the mitotic spindle to the kinetochores of paired sister chromatids in mitosis is monitored by a checkpoint that leads to an arrest in early metaphase. This arrest requires the inhibitory association of Mad2 with the anaphase promoting complex/cyclosome (APC/C). It is not known how the association of Mad2 with the kinetochore and the APC/C is regulated in mitosis. Here, we demonstrate that human Mad2 is modified through phosphorylation on multiple serine residues in vivo in a cell cycle dependent manner and that only unphosphorylated Mad2 interacts with Mad1 or the APC/C in vivo. A Mad2 mutant containing serine to aspartic acid mutations mimicking the C-terminal phosphorylation events fails to interact with Mad1 or the APC/C and acts as a dominant-negative antagonist of wild-type Mad2. These data suggest that the phosphorylation state of Mad2 regulates its checkpoint activity by modulating its association with Mad1 and the APC/C. PMID- 12574117 TI - Thylakoid targeting of Tat passenger proteins shows no delta pH dependence in vivo. AB - The Tat pathway is a major route for protein export in prokaryotes and for protein targeting to thylakoids in chloroplasts. Based on in vitro studies, protein translocation through this pathway is thought to be strictly dependent on a transmembrane delta pH. In this paper, we assess the delta pH sensitivity of the Tat pathway in vivo. Using Chlamydomonas reinhardtii, we observed changes in the efficiency of thylakoid targeting in vivo by mutating the Tat signal of the Rieske protein. We then employed two endogenous pH probes located on the lumen side of the thylakoid membranes to estimate spectroscopically the delta pH in vivo. Using experimental conditions in which the trans-thylakoid delta pH was almost zero, we found no evidence for a delta pH dependence of the Tat pathway in vivo. We confirmed this observation in higher plants using attached barley leaves. We conclude that the Tat pathway does not require a delta pH under physiological conditions, but becomes delta pH sensitive when probed in vitro/in organello because of the loss of some critical intracellular factors. PMID- 12574118 TI - Tim50, a novel component of the TIM23 preprotein translocase of mitochondria. AB - The preprotein translocase of the inner membrane of mitochondria (TIM23 complex) is the main entry gate for proteins of the matrix and the inner membrane. We isolated the TIM23 complex of Neurospora crassa. Besides Tim23 and Tim17, it contained a novel component, referred to as Tim50. Tim50 spans the inner membrane with a single transmembrane segment and exposes a large hydrophilic domain in the intermembrane space. Tim50 is essential for viability of yeast. Mitochondria from cells depleted of Tim50 displayed strongly reduced import kinetics of preproteins using the TIM23 complex. Tim50 could be cross-linked to preproteins that were halted at the level of the translocase of the outer membrane (TOM complex) or spanning both TOM and TIM23 complexes. We suggest that Tim50 plays a crucial role in the transfer of preproteins from the TOM complex to the TIM23 complex through the intermembrane space. PMID- 12574119 TI - Galpha(s) is palmitoylated at the N-terminal glycine. AB - Covalent lipid attachments are essential co- and post-translational modifications for signalling proteins. Galpha(s), the alpha-subunit of the heterotrimeric G protein that activates adenylyl cyclase, is known to be palmitoylated at the third N-terminal amino acid, a cysteine. Palmitoylation is involved in anchoring Galpha(s) to the membrane by increasing its intrinsic hydrophobicity. We identified by mass spectrometry a second, functionally even more important, covalent modification. It consists of another palmitoyl residue attached to the preceding glycine (Gly(2)). Palmitoylation at this position has profound consequences for levels of signal transduction. It sensitizes the cell up to 200 fold for adenylyl cyclase-stimulating agents. The inhibitory inputs mediated by Galpha(i) are downregulated to <10%. Thereby, Gly(2)-palmitoylation of Galpha(s) relieves cellular stimulation at the level of adenylyl cyclase whereas it renders the inhibitory modulation via Galpha(i) more difficult. PMID- 12574120 TI - Glycoprotein G isoforms from some alphaherpesviruses function as broad-spectrum chemokine binding proteins. AB - Mimicry of host chemokines and chemokine receptors to modulate chemokine activity is a strategy encoded by beta- and gammaherpesviruses, but very limited information is available on the anti-chemokine strategies encoded by alphaherpesviruses. The secretion of chemokine binding proteins (vCKBPs) has hitherto been considered a unique strategy encoded by poxviruses and gammaherpesviruses. We describe a family of novel vCKBPs in equine herpesvirus 1, bovine herpesvirus 1 and 5, and related alphaherpesviruses with no sequence similarity to chemokine receptors or other vCKBPs. We show that glycoprotein G (gG) is secreted from infected cells, binds a broad range of chemokines with high affinity and blocks chemokine activity by preventing their interaction with specific receptors. Moreover, gG also blocks chemokine binding to glycosaminoglycans, an interaction required for the correct presentation and function of chemokines in vivo. In contrast to other vCKBPs, gG may also be membrane anchored and, consistently, we show chemokine binding activity at the surface of cells expressing full-length protein. These alphaherpesvirus vCKBPs represent a novel family of proteins that bind chemokines both at the membrane and in solution. PMID- 12574121 TI - Association of Dishevelled with Eph tyrosine kinase receptor and ephrin mediates cell repulsion. AB - Eph tyrosine kinase receptors and their membrane-bound ligands, ephrins, are presumed to regulate cell-cell interactions. The major consequence of bidirectional activation of Eph receptors and ephrin ligands is cell repulsion. In this study, we discovered that Xenopus Dishevelled (Xdsh) forms a complex with Eph receptors and ephrin-B ligands and mediates the cell repulsion induced by Eph and ephrin. In vitro re-aggregation assays with Xenopus animal cap explants revealed that co-expression of a dominant-negative mutant of Xdsh affected the sorting of cells expressing EphB2 and those expressing ephrin-B1. Co-expression of Xdsh induced the activation of RhoA and Rho kinase in the EphB2-overexpressed cells and in the cells expressing EphB2-stimulated ephrin-B1. Therefore, Xdsh mediates both forward and reverse signaling of EphB2 and ephrin-B1, leading to the activation of RhoA and its effector protein Rho kinase. The inhibition of RhoA activity in animal caps significantly prevents the EphB2- and ephrin-B1 mediated cell sorting. We propose that Xdsh, which is expressed in various tissues, is involved in EphB and ephrin-B signaling related to regulation of cell repulsion via modification of RhoA activity. PMID- 12574122 TI - Mechanism of the spatio-temporal regulation of Ras and Rap1. AB - Epidermal growth factor (EGF) activates Ras and Rap1 at distinct intracellular regions. Here, we explored the mechanism underlying this phenomenon. We originally noticed that in cells expressing Epac, a cAMP-dependent Rap1 GEF (guanine nucleotide exchange factor), cAMP activated Rap1 at the perinuclear region, as did EGF. However, in cells expressing e-GRF, a recombinant cAMP responsive Ras GEF, cAMP activated Ras at the peripheral plasma membrane. Based on the uniform cytoplasmic expression of Epac and e-GRF, GEF did not appear to account for the non-uniform increase in the activities of Ras and Rap1. In contrast, when we used probes with reduced sensitivity to GTPase-activating proteins (GAPs), both Ras and Rap1 appeared to be activated uniformly in the EGF stimulated cells. Furthermore, we calculated the local rate constants of GEFs and GAPs from the video images of Ras activation and found that GAP activity was higher at the central plasma membrane than the periphery. Thus we propose that GAP primarily dictates the spatial regulation of Ras family G proteins, whereas GEF primarily determines the timing of Ras activation. PMID- 12574123 TI - Vibrio harveyi quorum sensing: a coincidence detector for two autoinducers controls gene expression. AB - In a process called quorum sensing, bacteria communicate with one another by exchanging chemical signals called autoinducers. In the bioluminescent marine bacterium Vibrio harveyi, two different auto inducers (AI-1 and AI-2) regulate light emission. Detection of and response to the V.harveyi autoinducers are accomplished through two two-component sensory relay systems: AI-1 is detected by the sensor LuxN and AI-2 by LuxPQ. Here we further define the V.harveyi quorum sensing regulon by identifying 10 new quorum-sensing-controlled target genes. Our examination of signal processing and integration in the V.harveyi quorum-sensing circuit suggests that AI-1 and AI-2 act synergistically, and that the V.harveyi quorum-sensing circuit may function exclusively as a 'coincidence detector' that discriminates between conditions in which both autoinducers are present and all other conditions. PMID- 12574124 TI - Recruitment of p300 by C/EBPbeta triggers phosphorylation of p300 and modulates coactivator activity. AB - Transcriptional coactivators such as p300 act as crucial elements in the eukaryotic gene regulation network. These proteins bind to various transcription factors which recruit them to specific gene regions whose chromatin structure subsequently is remodeled. Previously, we have shown that C/EBPbeta recruits p300 by interacting with the E1A-binding site of the coactivator. We now show that C/EBPbeta not only binds to p300 but also triggers massive phosphorylation of p300. This novel activity of C/EBPbeta is dependent on the E1A-binding region of p300 as well as on several subdomains of C/EBPbeta, all of which are involved in the p300-C/EBPbeta interaction. We have identified several sites of C/EBPbeta inducible phosphorylation within the C-terminal domain of p300. Mutation of these sites substantially impairs the activity of p300 as a coactivator of C/EBPbeta. Interestingly, phosphorylation of p300 is also triggered by other C/EBP family members as well as by various other transcription factors that interact with the E1A-binding domain of p300, suggesting that this novel phosphorylation mechanism may be of general relevance. PMID- 12574125 TI - STAT5-induced Id-1 transcription involves recruitment of HDAC1 and deacetylation of C/EBPbeta. AB - Transcriptional activation is associated commonly with recruitment of histone acetylases, while repression involves histone deacetylases (HDACs). Here, we provide evidence to suggest that STAT5 activates gene expression by recruiting HDAC. The interleukin-3 (IL-3)-dependent expression of the Id-1 gene, encoding a helix-loop-helix (HLH) transcriptional inhibitor, is activated by both C/EBPbeta and STAT5 transcription factors bound to its pro-B-cell enhancer (PBE), but is inhibited by HDAC inhibitors in Ba/F3 cells. STAT5 interacts with HDAC1 in the PBE region, resulting in deacetylation of histones, as well as C/EBPbeta, whose acetylation diminishes its DNA-binding activity. Consistently, expression of an acetylation-resistant mutant of C/EBPbeta results in IL-3-independent expression of the Id-1 gene. Thus, we propose a novel mechanism by which STAT5 mediates the deacetylation of C/EBPbeta, allowing transcriptional activation. PMID- 12574126 TI - A vertebrate RNA-binding protein Fox-1 regulates tissue-specific splicing via the pentanucleotide GCAUG. AB - Alternative splicing is one of the central mechanisms that regulate eukaryotic gene expression. Here we report a tissue-specific RNA-binding protein, Fox-1, which regulates alternative splicing in vertebrates. Fox-1 bound specifically to a pentanucleotide GCAUG in vitro. In zebrafish and mouse, fox-1 is expressed in heart and skeletal muscles. As candidates for muscle-specific targets of Fox-1, we considered two genes, the human mitochondrial ATP synthase gamma-subunit gene (F1gamma) and the rat alpha-actinin gene, because their primary transcripts contain several copies of GCAUG. In transfection experiments, Fox-1 induced muscle-specific exon skipping of the F1gamma gene via binding to GCAUG sequences upstream of the regulated exon. Fox-1 also regulated mutually exclusive splicing of the alpha-actinin gene, antagonizing the repressive effect of polypyrimidine tract-binding protein (PTB). It has been reported that GCAUG is essential for the alternative splicing regulation of several genes including fibronectin. We found that Fox-1 promoted inclusion of the fibronectin EIIIB exon. Thus, we conclude that Fox-1 plays key roles in both positive and negative regulation of tissue specific splicing via GCAUG. PMID- 12574127 TI - Isolation of a U-insertion/deletion editing complex from Leishmania tarentolae mitochondria. AB - A multiprotein, high molecular weight complex active in both U-insertion and U deletion as judged by a pre-cleaved RNA editing assay was isolated from mitochondrial extracts of Leishmania tarentolae by the tandem affinity purification (TAP) procedure, using three different TAP-tagged proteins of the complex. This editing- or E-complex consists of at least three protein-containing components interacting via RNA: the RNA ligase-containing L-complex, a 3' TUTase (terminal uridylyltransferase) and two RNA-binding proteins, Ltp26 and Ltp28. Thirteen approximately stoichiometric components were identified by mass spectrometric analysis of the core L-complex: two RNA ligases; homologs of the four Trypanosoma brucei editing proteins; and seven novel polypeptides, among which were two with RNase III, one with an AP endo/exonuclease and one with nucleotidyltransferase motifs. Three proteins have no similarities beyond kinetoplastids. PMID- 12574128 TI - The C-terminal domain of pol II and a DRB-sensitive kinase are required for 3' processing of U2 snRNA. AB - The human snRNA genes transcribed by RNA polymerase II (e.g. U1 and U2) have a characteristic TATA-less promoter containing an essential proximal sequence element. Formation of the 3' end of these non-polyadenylated RNAs requires a specialized 3' box element whose function is promoter specific. Here we show that truncation of the C-terminal domain (CTD) of RNA polymerase II and treatment of cells with CTD kinase inhibitors, including DRB (5,6-dichloro-1-beta-D ribofuranosylbenzimidazole), causes a dramatic reduction in proper 3' end formation of U2 transcripts. Activation of 3' box recognition by the phosphorylated CTD would be consistent with the role of phospho-CTD in mRNA processing. CTD kinase inhibitors, however, have little effect on initiation or elongation of transcription of the U2 genes, whereas elongation of transcription of the beta-actin gene is severely affected. This result highlights differences in transcription of snRNA and mRNA genes. PMID- 12574129 TI - Light-regulated translation mediates gated induction of the Arabidopsis clock protein LHY. AB - The transcription factor LHY and the related protein CCA1 perform overlapping functions in a regulatory feedback loop that is closely associated with the circadian oscillator of Arabidopsis: Overexpression of LHY abolished function of the circadian clock in constant light, but rhythmic expression of several circadian clock-regulated transcripts was observed under light- dark cycles. These oscillations correlated with high amplitude changes in LHY protein levels, caused by light-induced translation of the LHY transcript. Increases in LHY protein levels were also observed in light-grown wild-type plants, when light signals coincided with the circadian-regulated peak of LHY transcription at dawn. Unexpectedly, translational induction coincided with acute downregulation of LHY transcript levels. We suggest that the simultaneous translational induction and transcriptional repression of LHY expression play a role to narrow the peak of LHY protein synthesis at dawn and increase the robustness and accuracy of circadian oscillations. Strong phase shifting responses to light signals were observed in plants lacking function of LHY, CCA1 or both, suggesting that light regulated expression of these proteins does not mediate entrainment of the clock to light-dark cycles. PMID- 12574130 TI - Mechanism of Tet(O)-mediated tetracycline resistance. AB - Tet(O) is an elongation factor-like protein which confers resistance to the protein synthesis inhibitor tetracycline by promoting the release of the drug from its inhibitory site on the ribosome. Here we investigated the interaction of Tet(O) with the elongating ribosome and show, using dimethyl sulfate (DMS) probing and binding assays, that it interacts preferentially with the post translocational ribosome. Furthermore, using an XTP-dependent mutant of Tet(O), we demonstrated that Tet(O) induces conformational rearrangements within the ribosome which can be detected by EF-Tu, and manifested as a stimulation in the GTPase activity of this elongation factor. As such, these conformational changes probably involve the ribosomal GTPase-associated center and, accordingly, Tet(O) alters the DMS modification pattern of the L11 region. Additionally, tetracycline binding is associated with an E(a) of 58 kJ/mol. These results suggest a model where both Tet(O) and tetracycline induce a conformational change in functionally opposite directions and the Tet(O)-induced conformation persists after it has left the ribosome; this prevents rebinding of the drug while allowing productive A-site occupation by a ternary complex in the presence of tetracycline. PMID- 12574131 TI - E1 initiator DNA binding specificity is unmasked by selective inhibition of non specific DNA binding. AB - Initiator proteins are critical components of the DNA replication machinery and mark the site of initiation. This activity probably requires highly selective DNA binding; however, many initiators display modest specificity in vitro. We demonstrate that low specificity of the papillomavirus E1 initiator results from the presence of a non-specific DNA-binding activity, involved in melting, which masks the specificity intrinsic to the E1 DNA-binding domain. The viral factor E2 restores specificity through a physical interaction with E1 that suppresses non specific binding. We propose that this arrangement, where one DNA-binding activity tethers the initiator to ori while another alters DNA structure, is a characteristic of other viral and cellular initiator proteins. This arrangement would provide an explanation for the low selectivity observed for DNA binding by initiator proteins. PMID- 12574132 TI - Multiple ORC-binding sites are required for efficient MCM loading and origin firing in fission yeast. AB - In most eukaryotes, replication origins are composed of long chromosome regions, and the exact sequences required for origin recognition complex (ORC) and minichromosome maintenance (MCM) complex association remain elusive. Here, we show that two stretches of adenine/thymine residues are collectively essential for a fission yeast chromosomal origin. Chromatin immunoprecipitation assays revealed that the ORC subunits are located within a 1 kb region of ori2004. Analyses of deletion derivatives of ori2004 showed that adenine stretches are required for ORC binding in vivo. Synergistic interaction between ORC and adenine stretches was observed. On the other hand, MCM subunits were localized preferentially to a region near the initiation site, which is distant from adenine stretches. This association was dependent on adenine stretches and stimulated by a non-adenine element. Our results suggest that association of multiple ORC molecules with a replication origin is required for efficient MCM loading and origin firing in fission yeast. PMID- 12574136 TI - Transition from a continuous to discontinuous understanding of cardiac conduction. PMID- 12574133 TI - p53 is a chromatin accessibility factor for nucleotide excision repair of DNA damage. AB - One of the longest standing problems in DNA repair is how cells relax chromatin in order to make DNA lesions accessible for global nucleotide excision repair (NER). Since chromatin has to be relaxed for efficient lesion detection, the key question is whether chromatin relaxation precedes lesion detection or vice versa. Chromatin accessibility factors have been proposed but not yet identified. Here we show that p53 acts as a chromatin accessibility factor, mediating UV-induced global chromatin relaxation. Using localized subnuclear UV irradiation, we demonstrate that chromatin relaxation is extended over the whole nucleus and that this process requires p53. We show that the sequence for initiation of global NER is as follows: transcription-associated lesion detection; p53-mediated global chromatin relaxation; and global lesion detection. The tumour suppressor p53 is crucial for genomic stability, a role partially explained by its pro-apoptotic capacity. We demonstrate here that p53 is also a fundamental component of DNA repair, playing a direct role in rectifying DNA damage. PMID- 12574137 TI - Impaired hypoxic coronary vasodilation and ATP-sensitive potassium channel function: a manifestation of diabetic microangiopathy in humans? PMID- 12574138 TI - Serine phosphorylation and suppression of apoptosis by the small heat shock protein alphaB-crystallin. PMID- 12574139 TI - Onset of cardiac function during early mouse embryogenesis coincides with entry of primitive erythroblasts into the embryo proper. AB - When cardiac function and blood flow are first established are fundamental questions in mammalian embryogenesis. The earliest erythroblasts arise in yolk sac blood islands and subsequently enter the embryo proper to initiate circulation. Embryos staged 0 to 30 somites (S) were examined in utero with 40- to 50-MHz ultrasound biomicroscopy (UBM)-Doppler, to determine onset of embryonic heartbeat and blood flow and to characterize basic physiology of the very early mouse embryonic circulation. A heartbeat was first detected at 5 S, and blood vascular flow at 7 S. Heart rate, peak arterial velocity, and velocity-time integral showed progressive increases that indicated a dramatically increasing cardiac output from even the earliest stages. In situ hybridization revealed an onset of the heartbeat coincident with the appearance of yolk sac-derived erythroblasts in the embryo proper at 5 S. Early maturation of the circulation follows a tightly coordinated program. PMID- 12574140 TI - Beta-adrenergic receptor-stimulated apoptosis in cardiac myocytes is mediated by reactive oxygen species/c-Jun NH2-terminal kinase-dependent activation of the mitochondrial pathway. AB - Stimulation of beta-adrenergic receptors (betaARs) causes apoptosis in adult rat ventricular myocytes (ARVMs). The role of reactive oxygen species (ROS) in mediating betaAR-stimulated apoptosis is not known. Stimulation of betaARs with norepinephrine (10 micromol/L) in the presence of prazosin (100 nmol/L) for 24 hours increased the number of apoptotic myocytes as determined by TUNEL staining by 3.6- fold. The superoxide dismutase/catalase mimetics Mn(III)tetrakis(1-methyl 4-pyridyl)porphyrin pentachloride (MnTMPyP; 10 micromol/L) and Euk-134 decreased betaAR-stimulated apoptosis by 89+/-6% and 76+/-10%, respectively. Infection with an adenovirus expressing catalase decreased betaAR-stimulated apoptosis by 82+/ 15%. The mitochondrial permeability transition pore inhibitor bongkrekic acid (50 micromol/L) decreased betaAR-stimulated apoptosis by 76+/-8%, and the caspase inhibitor zVAD-fmk (25 micromol/L) decreased betaAR-stimulated apoptosis by 62+/ 11%. betaAR-stimulated cytochrome c release was inhibited by MnTMPyP. betaAR stimulation caused c-Jun NH2-terminal kinase (JNK) activation, which was abolished by MnTMPyP. Transfection with an adenovirus expressing dominant negative JNK inhibited betaAR-stimulated apoptosis by 81+/-12%, and the JNK inhibitor SP600125 inhibited both betaAR-stimulated apoptosis and cytochrome c release. Thus, betaAR-stimulated apoptosis in ARVMs involves ROS/JNK-dependent activation of the mitochondrial death pathway. PMID- 12574141 TI - Myocyte death, growth, and regeneration in cardiac hypertrophy and failure. AB - The accepted paradigm considers the adult mammalian heart as a postmitotic organ, which possesses a relatively constant number of myocytes from shortly after birth to adulthood and senescence. This notion is questioned by the demonstration that although most adult myocytes are terminally differentiated, there is a small and continuously renewed subpopulation of cycling myocytes produced by the differentiation of cardiac stem-like cells. Myocyte death and myocyte regeneration are introduced as major determinants of cardiac homeostasis and alterations of ventricular anatomy and function in physiological and pathological states. The possibility of reconstituting dead myocardium by stem-like cells is advanced and proposed as a major area of future research. PMID- 12574142 TI - Diabetes mellitus impairs vasodilation to hypoxia in human coronary arterioles: reduced activity of ATP-sensitive potassium channels. AB - ATP-sensitive K+ channels (K(ATP)) contribute to vasomotor regulation in some species. It is not fully understood the extent to which K(ATP) participate in regulating vasomotor tone under physiological and pathophysiological conditions in the human heart. Arterioles dissected from right atrial appendage were studied with video microscopy, membrane potential recordings, reverse transcription polymerase chain reaction, and immunohistochemistry. Hypoxia produced endothelium independent vasodilation and membrane hyperpolarization of vascular smooth muscle cells, both of which were attenuated by glibenclamide. Aprikalim, a selective K(ATP) opener, also induced a potent endothelium-independent and glibenclamide sensitive vasodilation with membrane hyperpolarization. Reverse transcription polymerase chain reaction detected mRNA expression for K(ATP) subunits, and immunohistochemistry confirmed the localization of the inwardly rectifying Kir6.1 protein in the vasculature. In patients with type 1 or type 2 diabetes mellitus (DM), vasodilation was reduced to both aprikalim (maximum dilation, DM(+) 90+/-2% versus DM(-) 96+/-1%, P<0.05) and hypoxia (maximum dilation, DM(+) 56+/-8% versus DM(-) 85+/-5%, P<0.01) but was not altered to sodium nitroprusside or bradykinin. Baseline myogenic tone and resting membrane potential were not affected by DM. We conclude that DM impairs human coronary arteriolar dilation to K(ATP) opening, leading to reduced dilation to hypoxia. This reduction in K(ATP) function could contribute to the greater cardiovascular mortality and morbidity in DM. PMID- 12574143 TI - Compound heterozygosity for mutations (W156X and R225W) in SCN5A associated with severe cardiac conduction disturbances and degenerative changes in the conduction system. AB - Cardiac conduction defects associate with mutations in SCN5A, the gene encoding the cardiac Na+ channel. In the present study, we characterized a family in which the proband was born in severe distress with irregular wide complex tachycardia. His older sister died at 1 year of age from severe conduction disease with similarly widened QRS-complexes. Mutational analysis of SCN5A in the proband demonstrated compound heterozygosity for a nonsense mutation (W156X), inherited from the father, and a missense mutation (R225W), inherited from the mother. Genotyping on DNA extracted from tissue from the deceased sibling revealed the same SCN5A genotype. Injection of cRNA encoding the W156X mutation in Xenopus oocytes did not produce any current. The R225W substitution neutralizes the third Arg residue within the voltage-sensing segment of domain I. Expression studies showed that this mutation leads to a severe reduction in I(Na) and is also associated with gating changes. Histological examination of the heart from the deceased sibling revealed changes consistent with a dilated type of cardiomyopathy and severe degenerative abnormalities of the specialized conduction system. The occurrence of compound heterozygosity for these two mutations implies that the proband carries solely severely dysfunctional cardiac Na+ channels. This explains his severe phenotype and that of his deceased sister who had been a carrier of the same genotype. The morphological changes within the heart of the deceased sibling may have occurred secondary to the Na+ channel abnormality and contributed to the severity of the disorder in this individual. PMID- 12574144 TI - Insulin-like growth factor-1 exerts Ca2+-dependent positive inotropic effects in failing human myocardium. AB - Myocardial generation of insulin-like growth factor-1 (IGF-1) is altered in hypertrophy and heart failure, but there are no reports on acute functional effects of IGF-1 in human cardiac muscle. We examined inotropic responses and signal transduction mechanisms of IGF-1 in human myocardium. Experiments were performed in isolated trabeculae or cardiomyocytes from 46 end-stage failing hearts. The effect of IGF-1 (0.001 to 0.2 micromol/L) on isometric twitch force (37 degrees C, 1 Hz), intracellular Ca2+ transients (aequorin method), sarcoplasmic reticulum (SR) Ca2+ content (rapid cooling contractures), L-type Ca2+ current (whole-cell voltage clamp), and cAMP concentrations was assessed. In addition, the effects of blocking IGF-1 receptors, phosphoinositide 3-kinase (PI3 kinase), protein kinase C (PKC), or transsarcolemmal Ca2+ entry were tested. IGF 1 exerted concentration-dependent positive inotropic effects (twitch force increased to maximally 133+/-4% of baseline values at 0.1 micromol/L; P<0.05). The IGF-1 receptor antibody alphaIR3 or the PI3-kinase inhibitor wortmannin prevented the functional effects. The inotropic response was paralleled by increases in Ca2+ transients and SR Ca2+ content. IGF-1 (0.1 micromol/L) increased L-type Ca2+ current amplitude by 24+/-7% (P<0.05). Blockade of SR function did not affect the inotropic response to IGF-1. In contrast, L-type Ca2+ channel blockade with diltiazem partially prevented ( approximately 50%) the inotropic response to IGF-1. Inhibition of PKC (GF109203X), Na+-H+ exchange (HOE642), or reverse-mode Na+-Ca2+ exchange (KB-R7943) reduced the response to IGF-1 by approximately 60% to 70%. IGF-1 exerts Ca2+-dependent positive inotropic effects through activation of IGF-1 receptors and a PI3-kinase-dependent pathway in failing human myocardium. The increased [Ca2+]i with IGF-1 originates from both enhanced L-type Ca2+ currents and enhanced Na+-H+ exchange-dependent reverse mode Na+-Ca2+ exchange. These nongenomic functional effects of IGF-1 may be of clinical relevance. PMID- 12574145 TI - Selective matrix metalloproteinase inhibition with developing heart failure: effects on left ventricular function and structure. AB - The matrix metalloproteinases (MMPs) are an endogenous family of proteolytic enzymes implicated to contribute to LV remodeling. However, broad-spectrum MMP inhibition (MMPi), particularly inhibition of interstitial collagenase (MMP-1), may not be clinically applicable. This study examined the effects of selective MMPi (sparing MMP-1) in a model of developing congestive heart failure. Pigs were randomly assigned to 3 groups: (1) rapid pacing for 3 weeks (240 bpm, n=10); (2) selective MMPi (20 mg/kg per day-PO;PGE7113313) and rapid pacing (n=12); and (3) controls (n=10). LV peak wall stress increased from controls with rapid pacing (140+/-6 versus 319+/-18 g/cm2; P<0.05) and was reduced with selective MMPi (208+/-9 g/cm2; P<0.05. Preload recruitable stroke work was reduced with rapid pacing (4.3+/-0.4 versus 1.2+/-0.2 dyne. cm/mm Hg; P<0.05) and was increased with selective MMPi (2.6+/-0.3 dyne. cm/mm Hg; P<0.05). Plasma norepinephrine increased by 6-fold in the rapid pacing group (P<0.05) and was reduced from untreated values with selective MMPi (P<0.05). At the myocardial level, myocyte cross-sectional area was increased with selective MMPi but fibrillar collagen volume fraction remained unchanged relative to control values. These results suggest that targeting a selective portfolio of myocardial MMP species for inhibition may provide a more rational therapeutic strategy in the setting of congestive heart failure. PMID- 12574146 TI - Mechanistically distinct steps in the mitochondrial death pathway triggered by oxidative stress in cardiac myocytes. AB - Oxidative stress plays an important role in the pathogenesis of cardiovascular diseases. In the present study, we characterize three distinct phases of the H2O2 induced response, which leads to loss of mitochondrial membrane potential (DeltaPsi(m)) and subsequent cell death in cultured cardiac myocytes. (1) Priming: After H2O2 exposure (100 micromol/L), cells maintain a constant DeltaPsi(m) for the cell-to-cell specific latency but at the same time undergo progressive changes in inner mitochondrial membrane structure (swelling and loss of cristae by electron microscopy). An increase of matrix calcium is required, but not sufficient, for this process. (2) Depolarization: Priming is followed by sudden depolarization of DeltaPsi(m), which is mediated by mitochondrial permeability transition pore opening, as evidenced by the concomitant release of calcein from mitochondria. This process is rapid (<4 minutes), complete, and irreversible. The duration of depolarization is constant and does not depend on the length of the priming process in any given cell. (3) Fragmentation: Along with massive mitochondrial swelling and release of cytochrome c into the cytoplasm, cells undergo surface membrane alterations, such as exposure of phosphatidylserine and eventual loss of membrane integrity and cellular fragmentation. Thus, oxidant stress elicits reproducible and stereotyped responses in cardiac cells. The priming phase, during which mitochondria undergo major ultrastructural alterations but remain functional, represents a particularly attractive target for intervention in the prevention of cell death. PMID- 12574147 TI - Differential actions of cardioprotective agents on the mitochondrial death pathway. AB - We examined the effect of cardioprotective agents on three distinct phases of the H2O2-induced response that leads to loss of mitochondrial membrane potential (DeltaPsi(m)) and cell death in cultured cardiac myocytes: (1) priming, consisting of calcium-dependent morphological changes in mitochondria (swelling and loss of cristae), with preserved DeltaPsi(m), (2) depolarization, the rapid DeltaPsi(m) depolarization caused by mitochondrial permeability transition pore (PTP) opening, and (3) cell fragmentation. The mitochondrial ATP-sensitive potassium (mitoK(ATP)) channel opener diazoxide markedly decreased the likelihood that cells would undergo priming: many mitochondria remained fully polarized and morphologically intact. Diazoxide not only decreased the number of cells undergoing DeltaPsi(m) depolarization but also delayed the onset of DeltaPsi(m) loss, whereas it did not change the duration of depolarization in unprotected cells. The adenine nucleotide translocase inhibitor bongkrekic acid mimicked the effect of diazoxide to suppress priming, except that its effects were not blocked by the mitoK(ATP) channel blocker 5-hydroxydecanoate. In contrast, the PTP inhibitor cyclosporin A (CsA) did not prevent priming: neither latency for DeltaPsi(m) depolarization nor mitochondrial morphological changes were affected. However, CsA slowed the process of depolarization and blunted its severity. Importantly, coapplication of diazoxide and CsA exhibited additive effects, improving the efficacy of protection. Activation of mitoK(ATP) channels suppresses the cell death process at its earliest stage, by preserving mitochondrial integrity during oxidative stress. By virtue of its pharmacology and its phenotypic consequences, this mode of action is distinguishable from that of other cardioprotective interventions. PMID- 12574148 TI - Mimicking phosphorylation of alphaB-crystallin on serine-59 is necessary and sufficient to provide maximal protection of cardiac myocytes from apoptosis. AB - AlphaB-crystallin (alphaBC), a small heat shock protein expressed in high levels in the heart, is phosphorylated on Ser-19, 45, and 59 after stress. However, it is not known whether alphaBC phosphorylation directly affects cell survival. In the present study, constructs were prepared that encode forms of alphaBC harboring Ser to Ala (blocks phosphorylation) or Ser to Glu (mimics phosphorylation) mutations at positions 19, 45, and 59. The effects of each form on apoptosis of cultured cardiac myocytes after hyperosmotic or hypoxic stress were assessed. Compared with controls, cells that expressed alphaBC with Ser to Ala substitutions at all three positions, alphaBC(AAA), exhibited more stress induced apoptosis. Cells expressing either alphaBC(AAE) or (EEE) exhibited 3-fold less apoptosis than cells expressing alphaBC(AAA), indicating that phosphorylation of Ser-59 confers protection. alphaBC is known to bind to procaspase-3 and to decrease caspase-3 activation. Compared with cells expressing alphaBC(AAA), the activation of caspase-3 was decreased by 3-fold in cells expressing alphaBC(AAE). These results demonstrate that mimicking the phosphorylation of alphaBC on Ser-59 is necessary and sufficient to confer caspase-3 inhibition and protection of cardiac myocytes against hyperosmotic or hypoxic stress. These findings provide direct evidence that alphaBC(S59P) contributes to the cardioprotection observed after physiologically relevant stresses, such as transient hypoxia. Identifying the targets of alphaBC(S59P) will reveal important details about the mechanism underlying the cytoprotective effects of this small heat shock protein. PMID- 12574149 TI - Peroxisome proliferator-activated receptor alpha reduces cholesterol esterification in macrophages. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a nuclear receptor activated by fatty acid derivatives and hypolipidemic drugs of the fibrate class. PPARalpha is expressed in monocytes, macrophages, and foam cells, suggesting a role for this receptor in macrophage lipid homeostasis with consequences for atherosclerosis development. Recently, it was shown that PPARalpha activation promotes cholesterol efflux from macrophages via induction of the ABCA1 pathway. In the present study, the influence of PPARalpha activators on intracellular cholesterol homeostasis was investigated. In human macrophages and foam cells, treatment with fibrates, synthetic PPARalpha activators, led to a decrease in the cholesteryl ester (CE):free cholesterol (FC) ratio. In these cells, PPARalpha activation reduced cholesterol esterification rates and Acyl CoA:cholesterol acyltransferase-1 (ACAT1) activity. However, PPARalpha activation did not alter ACAT1 gene expression, whereas mRNA levels of carnitine palmitoyltransferase type 1 (CPT-1), a key enzyme in mitochondrial fatty acid catabolism, were induced. Finally, PPARalpha activation blocked CE formation induced by TNF-alpha, possibly due to the inhibition of neutral sphingomyelinase activation by TNF-alpha. In conclusion, our results identify a role for PPARalpha in the control of cholesterol esterification in macrophages, resulting in an enhanced availability of FC for efflux through the ABCA1 pathway. PMID- 12574150 TI - Local monocyte chemoattractant protein-1 therapy increases collateral artery formation in apolipoprotein E-deficient mice but induces systemic monocytic CD11b expression, neointimal formation, and plaque progression. AB - Monocyte chemoattractant protein-1 (MCP-1) stimulates the formation of a collateral circulation on arterial occlusion. The present study served to determine whether these proarteriogenic properties of MCP-1 are preserved in hyperlipidemic apolipoprotein E-deficient (apoE-/-) mice and whether it affects the systemic development of atherosclerosis. A total of 78 apoE-/- mice were treated with local infusion of low-dose MCP-1 (1 microg/kg per week), high-dose MCP-1 (10 microg/kg per week), or PBS as a control after unilateral ligation of the femoral artery. Collateral hindlimb flow, measured with fluorescent microspheres, significantly increased on a 1-week high-dose MCP-1 treatment (PBS 22.6+/-7.2%, MCP-1 31.3+/-10.3%; P<0.05). These effects were still present 2 months after the treatment (PBS 44.3+/-4.6%, MCP-1 56.5+/-10.4%; P<0.001). The increase in collateral flow was accompanied by an increase in the number of perivascular monocytes/macrophages on MCP-1 treatment. However, systemic CD11b expression by monocytes also increased, as did monocyte adhesion at the aortic endothelium and neointimal formation (intima/media ratio, 0.097+/-0.011 [PBS] versus 0.257+/-0.022 [MCP-1]; P<0.0001). Moreover, Sudan IV staining revealed an increase in aortic atherosclerotic plaque surface (24.3+/-5.2% [PBS] versus 38.2+/-9.5% [MCP-1]; P<0.01). Finally, a significant decrease in the percentage of smooth muscle cells was found in plaques (15.0+/-5.2% [PBS] versus 5.8+/-2.3% [MCP-1]; P<0.001). In conclusion, local infusion of MCP-1 significantly increases collateral flow on femoral artery ligation in apoE-/- mice up to 2 months after the treatment. However, the local treatment did not preclude systemic effects on atherogenesis, leading to increased atherosclerotic plaque formation and changes in cellular content of plaques. PMID- 12574151 TI - Regulation of cytokine-induced nitric oxide synthesis by asymmetric dimethylarginine: role of dimethylarginine dimethylaminohydrolase. AB - In response to vascular insults, inflammatory cytokines stimulate vascular smooth muscle cells (SMCs) to express an inducible isoform of nitric oxide synthase (iNOS). Asymmetric dimethylarginine (ADMA), an endogenous NO synthase inhibitor, is metabolized by dimethylarginine dimethylaminohydrolase (DDAH). To determine whether the ADMA-DDAH system regulates cytokine-induced NO production, cultured rat SMCs were exposed to interleukin-1beta (IL-1beta). IL-1beta (1 to 100 U/mL) dose-dependently stimulated not only iNOS but also DDAH expression and enzyme activity, accompanied by an increase in NO metabolite and by a decrease in ADMA content in culture media. A DDAH inhibitor (4124W, 5 mmol/L) augmented ADMA production (P<0.01) and decreased NO synthesis (P<0.01) in IL-1beta-stimulated SMCs. On the other hand, an adenovirus-mediated overexpression of DDAH reduced ADMA and enhanced NO production. Exogenous administration of NO donors (SNAP and SIN-1) dose-dependently increased NO metabolite in the culture media but had no effect on ADMA. Our results indicate two mechanisms of IL-1beta-induced NO synthesis: the direct stimulation of the expression of iNOS and the indirect stimulation of iNOS activity by upregulating DDAH and reducing ADMA. The ADMA DDAH system may be another regulatory mechanism of inflammation-mediated NO production for human vascular diseases. PMID- 12574152 TI - Microarray analysis reveals complex remodeling of cardiac ion channel expression with altered thyroid status: relation to cellular and integrated electrophysiology. AB - Although electrophysiological remodeling occurs in various myocardial diseases, the underlying molecular mechanisms are poorly understood. cDNA microarrays containing probes for a large population of mouse genes encoding ion channel subunits ("IonChips") were developed and exploited to investigate remodeling of ion channel transcripts associated with altered thyroid status in adult mouse ventricle. Functional consequences of hypo- and hyperthyroidism were evaluated with patch-clamp and ECG recordings. Hypothyroidism decreased heart rate and prolonged QTc duration. Opposite changes were observed in hyperthyroidism. Microarray analysis revealed that hypothyroidism induces significant reductions in KCNA5, KCNB1, KCND2, and KCNK2 transcripts, whereas KCNQ1 and KCNE1 expression is increased. In hyperthyroidism, in contrast, KCNA5 and KCNB1 expression is increased and KCNQ1 and KCNE1 expression is decreased. Real-time RT-PCR validated these results. Consistent with microarray analysis, Western blot experiments confirmed those modifications at the protein level. Patch-clamp recordings revealed significant reductions in I(to,f) and I(K,slow) densities, and increased I(Ks) density in hypothyroid myocytes. In addition to effects on K+ channel transcripts, transcripts for the pacemaker channel HCN2 were decreased and those encoding the alpha1C Ca2+ channel (CaCNA1C) were increased in hypothyroid animals. The expression of Na+, Cl-, and inwardly rectifying K+ channel subunits, in contrast, were unaffected by thyroid hormone status. Taken together, these data demonstrate that thyroid hormone levels selectively and differentially regulate transcript expression for at least nine ion channel alpha- and beta subunits. Our results also document the potential of cDNA microarray analysis for the simultaneous examination of ion channel transcript expression levels in the diseased/remodeled myocardium. PMID- 12574153 TI - Load-induced transcriptional activation of c-jun in rat myocardium: regulation by myocyte enhancer factor 2. AB - The increased expression of immediate-early genes is a key feature of the myocardial response to hypertrophic stimuli. In this study, we investigated whether pressure overload or phenylephrine treatment stimulated myocyte enhancer factor 2 (MEF2)-dependent transcriptional activation of c-jun in cardiac myocytes. Western blotting and immunohistochemical analysis of rat myocardium demonstrated that p70(MEF2) is highly expressed in the rat heart and is predominantly located at the nuclei of cardiac myocytes. Electrophoretic mobility shift assays of myocardial nuclear extracts revealed a consistent DNA binding activation of MEF2 after 1 and 2 hours of pressure overload. We further showed that pressure overload induced a progressive nuclear translocation and activation of extracellular signal-regulated kinase 5 (ERK5). Coimmunoprecipitation and in vitro kinase assays indicated that the activation of ERK5 was paralleled by increased association of ERK5/p70(MEF2) and by enhanced ability of ERK5 to phosphorylate p70(MEF2). Experiments with in vivo transfection of the left ventricle with the c-jun promoter reporter gene showed that pressure overload induced a consistent increase of c-jun transcriptional activity in the rat myocardium. Rendering the MEF2 site of the c-jun plasmid inactive by mutation abolished the load-induced activation of the c-jun promoter reporter gene. Mutation of the MEF2 site also abolished the phenylephrine-induced c-jun promoter activation in neonatal rat ventricular myocytes. In addition, we demonstrated that neonatal rat ventricular myocyte transfection with ERK5-antisense oligodeoxynucleotide inhibited the phenylephrine-induced c-jun promoter activation. These findings identify MEF2 as a potential regulator of c-jun transactivation and suggest that ERK5 might be an important mediator of MEF2 and c-jun promoter activation in response to hypertrophic stimuli in cardiac myocytes. PMID- 12574154 TI - Role for hydrogen peroxide in flow-induced dilation of human coronary arterioles. AB - Flow-induced dilation (FID) is dependent largely on hyperpolarization of vascular smooth muscle cells (VSMCs) in human coronary arterioles (HCA) from patients with coronary disease. Animal studies show that shear stress induces endothelial generation of hydrogen peroxide (H2O2), which is proposed as an endothelium derived hyperpolarizing factor (EDHF). We tested the hypothesis that H2O2 contributes to FID in HCA. Arterioles (135+/-7 micro m, n=71) were dissected from human right atrial appendages at the time of cardiac surgery and cannulated with glass micropipettes. Changes in internal diameter and membrane potential of VSMCs to shear stress, H2O2, or to papaverine were recorded with videomicroscopy. In some vessels, endothelial H2O2 generation to shear stress was monitored directly using confocal microscopy with 2',7'-dichlorofluorescin diacetate (DCFH) or using electron microscopy with cerium chloride. Catalase inhibited FID (%max dilation; 66+/-8 versus 25+/-7%; P<0.05, n=6), whereas dilation to papaverine was unchanged. Shear stress immediately increased DCFH fluorescence in the endothelial cell layer, whereas treatment with catalase abolished the increase in fluorescence. Electron microscopy with cerium chloride revealed shear stress induced increase in cerium deposition in intimal area surrounding endothelial cells. Exogenous H2O2 dilated (%max dilation; 97+/-1%, ED50; 3.0+/-0.7x10(-5) mol/L) and hyperpolarized HCA. Dilation to H2O2 was reduced by catalase, 40 mmol/L KCl, or charybdotoxin plus apamin, whereas endothelial denudation, deferoxamine, 1H-(1,2,4)-oxadiazole-[4,3-a]quinoxalin-1-one, or glibenclamide had no effect. These data provide evidence that shear stress induces endothelial release of H2O2 and are consistent with the idea that H2O2 is an EDHF that contributes to FID in HCA from patients with heart disease. The full text of this article is available at http://www.circresaha.org. PMID- 12574155 TI - Deletion of selenoprotein P alters distribution of selenium in the mouse. AB - Selenoprotein P (Se-P) contains most of the selenium in plasma. Its function is not known. Mice with the Se-P gene deleted (Sepp(-/-)) were generated. Two phenotypes were observed: 1) Sepp(-/-) mice lost weight and developed poor motor coordination when fed diets with selenium below 0.1 mg/kg, and 2) male Sepp(-/-) mice had sharply reduced fertility. Weanling male Sepp(+/+), Sepp(+/-), and Sepp( /-) mice were fed diets for 8 weeks containing <0.02-2 mg selenium/kg. Sepp(+/+) and Sepp(+/-) mice had similar selenium concentrations in all tissues except plasma where a gene-dose effect on Se-P was observed. Liver selenium was unaffected by Se-P deletion except that it increased when dietary selenium was below 0.1 mg/kg. Selenium in other tissues exhibited a continuum of responses to Se-P deletion. Testis selenium was depressed to 19% in mice fed an 0.1 mg selenium/kg diet and did not rise to Sepp(+/+) levels even with a dietary selenium of 2 mg/kg. Brain selenium was depressed to 43%, but feeding 2 mg selenium/kg diet raised it to Sepp(+/+) levels. Kidney was depressed to 76% and reached Sepp(+/+) levels on an 0.25 mg selenium/kg diet. Heart selenium was not affected. These results suggest that the Sepp(-/-) phenotypes were caused by low selenium in testis and brain. They strongly suggest that Se-P from liver provides selenium to several tissues, especially testis and brain. Further, they indicate that transport forms of selenium other than Se-P exist because selenium levels of all tissues except testis responded to increases of dietary selenium in Sepp(-/-) mice. PMID- 12574156 TI - Melanoma cell CD44 interaction with the alpha 1(IV)1263-1277 region from basement membrane collagen is modulated by ligand glycosylation. AB - Invasion of the basement membrane is believed to be a critical step in the metastatic process. Melanoma cells have been shown previously to bind distinct triple-helical regions within basement membrane (type IV) collagen. Additionally, tumor cell binding sites within type IV collagen contain glycosylated hydroxylysine residues. In the present study, we have utilized triple-helical models of the type IV collagen alpha1(IV)1263-1277 sequence to (a) determine the melanoma cell receptor for this ligand and (b) analyze the results of single-site glycosylation on melanoma cell recognition. Receptor identification was achieved by a combination of methods, including (a) cell adhesion and spreading assays using triple-helical alpha1(IV)1263-1277 and an Asp(1266)Abu variant, (b) inhibition of cell adhesion and spreading assays, and (c) triple-helical alpha1(IV)1263-1277 affinity chromatography with whole cell lysates and glycosaminoglycans. Triple-helical alpha1(IV)1263-1277 was bound by melanoma cell CD44/chondroitin sulfate proteoglycan receptors and not by the collagen-binding integrins or melanoma-associated proteoglycan. Melanoma cell adhesion to and spreading on the triple-helical alpha1(IV)1263-1277 sequence was then compared for glycosylated (replacement of Lys(1265) with Hyl(O-beta-d-galactopyranosyl)) versus non-glycosylated ligand. Glycosylation was found to strongly modulate both activities, as adhesion and spreading were dramatically decreased due to the presence of galactose. CD44/chondroitin sulfate proteoglycan did not bind to glycosylated alpha1(IV)1263-1277. Overall, this study (a) is the first demonstration of the prophylactic effects of glycosylation on tumor cell interaction with the basement membrane, (b) provides a rare example of an apparent unfavorable interaction between carbohydrates, and (c) suggests that sugars may mask "cryptic sites" accessible to tumor cells with cell surface or secreted glycosidase activities. PMID- 12574158 TI - The extracellular domain of receptor activity-modifying protein 1 is sufficient for calcitonin receptor-like receptor function. AB - A functional calcitonin gene-related peptide (CGRP) receptor requires dimerization of calcitonin receptor-like receptor (CRLR) with receptor activity modifying protein 1 (RAMP 1). To determine the function of the three domains (extracellular, ECD; transmembrane, TM; and tail domains) of human RAMP 1, three mutants were constructed: RAMP 1 without the cytoplasmic tail, a chimera consisting of the ECD of RAMP 1 and the TM and tail of the platelet-derived growth factor receptor, and the ECD of RAMP 1 alone. These RAMP 1 mutants were examined for their ability to associate with CRLR to effect CGRP-stimulated cAMP accumulation, CGRP binding, CRLR trafficking, and cell surface expression. All RAMP 1 mutants were able to associate with CRLR with full efficacy for CGRP stimulated cAMP accumulation. However, the RAMP 1/platelet-derived growth factor receptor chimera demonstrated a 10-fold decrease in potency for CGRP signaling and binding, and the RAMP 1-ECD mutant had a 4000-fold decrease in potency. In conclusion, the ECD of RAMP 1 is sufficient for normal CRLR association and efficacy. The presence of a TM domain and the specific sequence of the RAMP 1 TM domain contribute to CGRP affinity and potency. The C-terminal tail of RAMP 1 is unnecessary for CRLR function. PMID- 12574157 TI - Interactions of mouse Paneth cell alpha-defensins and alpha-defensin precursors with membranes. Prosegment inhibition of peptide association with biomimetic membranes. AB - The bactericidal activity of mouse alpha-defensins (cryptdins) requires proteolytic activation of inactive precursors by matrix metalloproteinase-7 (matrilysin, EC, MMP-7(a)). To investigate mechanisms of cryptdin-4 (Crp4) peptide interactions with membrane bilayers and to determine whether MMP-7 mediated proteolysis activates the membrane disruptive activity of Crp4, associations of Crp4 and melittin with biomimetic lipid/polydiacetylene chromatic vesicles were characterized. The peptides differ in their sensitivity to vesicle lipid composition and their depth of bilayer penetration. Crp4 undergoes strong interfacial binding onto lipid bilayers with disruption of the bilayer head group region, unlike melittin, which inserts more deeply into the hydrophobic core of the bilayer. Colorimetric and tryptophan fluorescence studies showed that Crp4 insertion is favored by negatively charged phospholipids and that zwitterionic and Escherichia coli phospholipids promote stronger interfacial binding; melittin membrane interactions were independent of either variable. In contrast to the membrane disruptive activity of Crp4, pro-Crp4 did not perturb vesicular membranes, consistent with the lack of bactericidal activity of the precursor, and incubation of Crp4 with prosegment in trans blocked Crp4 and G1W-Crp4 membrane interactions at concentrations that inhibit Crp4 bactericidal activity. CD measurements showed that Crp4 has an expected beta-sheet structure that is not evident in the pro-Crp4 CD trace or when Crp4 is incubated with prosegment, indicating that the beta-sheet signal is attenuated by proregion interactions or possibly disrupted by the prosegment. Collectively, the results suggest that the prosegment inhibits Crp4 bactericidal activity by blocking peptide-mediated perturbation of target cell membranes, a constraint that is relieved when MMP-7 cleaves the prosegment. PMID- 12574159 TI - Rapid induction of cell death by selenium-compromised thioredoxin reductase 1 but not by the fully active enzyme containing selenocysteine. AB - Mammalian thioredoxin reductases are selenoproteins. For native catalytic activity, these enzymes utilize a C-terminal -Gly-Cys-Sec-Gly-COOH sequence (where Sec is selenocysteine) forming a redox active selenenylsulfide/selenolthiol motif. A range of cellular systems depend upon or are regulated by thioredoxin reductase and its major protein substrate thioredoxin, including apoptosis signal-regulating kinase 1, peroxiredoxins, methionine sulfoxide reductase, and several transcription factors. Cytosolic thioredoxin reductase 1 (TrxR1) is moreover inhibited by various electrophilic anticancer compounds. TrxR1 is hence generally considered to promote cell viability. However, several recent studies have suggested that TrxR1 may promote apoptosis, and the enzyme was identified as GRIM-12 (gene associated with retinoid interferon-induced mortality 12). Transient transfection with GRIM 12/TrxR1 was also shown to directly induce cell death. To further analyze such effects, we have here employed lipid-mediated delivery of recombinant TrxR1 preparations into human A549 cells, thereby bypassing selenoprotein translation to facilitate assessment of the protein-related effects on cell viability. We found that selenium-deficient TrxR1, having a two-amino acid-truncated C-terminal -Gly-Cys-COOH motif, rapidly induced cell death (38 +/- 29% apoptotic cells after 4 h; p < 0.005 compared with controls). Cell death induction was also promoted by selenium-compromised TrxR1 derivatized with either cis-diamminedichloroplatinum (II) (cisplatin) or dinitrophenyl moieties but not by the structurally related non-selenoprotein glutathione reductase. In contrast, TrxR1 with intact selenocysteine could not promote cell death. The direct cellular effects of selenium-compromised forms of TrxR1 may be important for the pathophysiology of selenium deficiency as well as for the efficacy of antiproliferative drugs targeting the selenocysteine moiety of this enzyme. PMID- 12574160 TI - Trafficking patterns of beta-arrestin and G protein-coupled receptors determined by the kinetics of beta-arrestin deubiquitination. AB - Agonist-dependent internalization of G protein-coupled receptors via clathrin coated pits is dependent on the adaptor protein beta-arrestin, which interacts with elements of the endocytic machinery such as AP2 and clathrin. For the beta(2)-adrenergic receptor (beta(2)AR) this requires ubiquitination of beta arrestin by E3 ubiquitin ligase, Mdm2. Based on trafficking patterns and affinity of beta-arrestin, G protein-coupled receptors are categorized into two classes. For class A receptors (e.g. beta(2)AR), which recycle rapidly, beta-arrestin directs the receptors to clathrin-coated pits but does not internalize with them. For class B receptors (e.g. V2 vasopressin receptors), which recycle slowly, beta arrestin internalizes with the receptor into endosomes. In COS-7 and human embryonic kidney (HEK)-293 cells, stimulation of the beta(2)AR or V2 vasopressin receptor leads, respectively, to transient or stable beta-arrestin ubiquitination. The time course of ubiquitination and deubiquitination of beta arrestin correlates with its association with and dissociation from each type of receptor. Chimeric receptors, constructed by switching the cytoplasmic tails of the two classes of receptors (beta(2)AR and V2 vasopressin receptors), demonstrate reversal of the patterns of both beta-arrestin trafficking and beta arrestin ubiquitination. To explore the functional consequences of beta-arrestin ubiquitination we constructed a yellow fluorescent protein-tagged beta-arrestin2 ubiquitin chimera that cannot be deubiquitinated by cellular deubiquitinases. This "permanently ubiquitinated" beta-arrestin did not dissociate from the beta(2)AR but rather internalized with it into endosomes, thus transforming this class A receptor into a class B receptor with respect to its trafficking pattern. Overexpression of this beta-arrestin ubiquitin chimera in HEK-293 cells also results in enhancement of beta(2)AR internalization and degradation. In the presence of N-ethylmaleimide (an inhibitor of deubiquitinating enzymes), coimmunoprecipitation of the receptor and beta-arrestin was increased dramatically, suggesting that deubiquitination of beta-arrestin triggers its dissociation from the receptor. Thus the ubiquitination status of beta-arrestin determines the stability of the receptor-beta-arrestin complex as well as the trafficking pattern of beta-arrestin. PMID- 12574161 TI - PNUTS, a protein phosphatase 1 (PP1) nuclear targeting subunit. Characterization of its PP1- and RNA-binding domains and regulation by phosphorylation. AB - PNUTS, Phosphatase 1 NUclear Targeting Subunit, is a recently described protein that targets protein phosphatase 1 (PP1) to the nucleus. In the present study, we characterized the biochemical properties of PNUTS. A variety of truncation and site-directed mutants of PNUTS was prepared and expressed either as glutathione S transferase fusion proteins in Escherichia coli or as FLAG-tagged proteins in 293T cells. A 50-amino acid domain in the center of PNUTS mediated both high affinity PP1 binding and inhibition of PP1 activity. The PP1-binding domain is related to a motif found in several other PP1-binding proteins but is distinct in that Trp replaces Phe. Mutation of the Trp residue essentially abolished the ability of PNUTS to bind to and inhibit PP1. The central PP1-binding domain of PNUTS was an effective substrate for protein kinase A in vitro, and phosphorylation substantially reduced the ability of PNUTS to bind to PP1 in vitro and following stimulation of protein kinase A in intact cells. In vitro RNA binding experiments showed that a C-terminal region including several RGG motifs and a novel repeat domain rich in His and Gly interacted with mRNA and single stranded DNA. PNUTS exhibited selective binding for poly(A) and poly(G) compared with poly(U) or poly(C) ribonucleotide homopolymers, with specificity being mediated by distinct regions within the domain rich in His and Gly and the domain containing the RGG motifs. Finally, a PNUTS-PP1 complex was isolated from mammalian cell lysates using RNA-conjugated beads. Together, these studies support a role for PNUTS in protein kinase A-regulated targeting of PP1 to specific RNA-associated complexes in the nucleus. PMID- 12574162 TI - The requirement for polyamines for intestinal epithelial cell migration is mediated through Rac1. AB - The rapid migration of intestinal epithelial cells is important to the healing of mucosal ulcers and wounds. This cell migration requires the presence of polyamines and the activation of RhoA. RhoA activity, however, is not sufficient for migration because polyamine depletion inhibited the migration of IEC-6 cells expressing constitutively active RhoA. The current study examines the role of Rac1 and Cdc42 in cell migration and whether their activities are polyamine dependent. Polyamine depletion with alpha-difluoromethylornithine inhibited the activities of RhoA, Rac1, and Cdc42. This inhibition was prevented by supplying exogenous putrescine in the presence of alpha-difluoromethylornithine. IEC-6 cells transfected with constitutively active Rac1 and Cdc42 migrated more rapidly than vector-transfected cells, whereas cells expressing dominant negative Rac1 and Cdc42 migrated more slowly. Polyamine depletion had no effect on the migration of cells expressing Rac1 and only partially inhibited the migration of those expressing Cdc42. Although polyamine depletion caused the disappearance of actin stress fibers in cells transfected with empty vector, it had no effect on cells expressing Rac1. Constitutively active Rac1 increased RhoA and Cdc42 activity in both normal and polyamine-depleted cells. These results demonstrate that Rac1, RhoA, and Cdc42 are required for optimal epithelial cell migration and that Rac1 activity is sufficient for cell migration in the absence of polyamines due to its ability to activate RhoA and Cdc42 as well as its own effects on the process of cell migration. These data imply that the involvement of polyamines in cell migration occurs either at Rac1 itself or upstream from Rac1. PMID- 12574163 TI - Identification and characterization of a novel Ste20/germinal center kinase related kinase, polyploidy-associated protein kinase. AB - A novel protein kinase, polyploidy-associated protein kinase (PAPK), was isolated using a subtraction cDNA library approach from a mouse erythroleukemia cell line that had been induced to polyploidy after serum withdrawal. PAPK shares homology with members of the Ste20/germinal center kinase family of protein kinases and is ubiquitously expressed as two spliced forms, PAPK-A and PAPK-B, that encode for proteins of 418 and 189 amino acids, respectively. The expression of endogenous PAPK-A protein increased after growth factor withdrawal in murine hematopoietic and fibroblast cells. When tested in an in vitro kinase assay, PAPK-A was activated in response to the stress-inducing agent hydrogen peroxide and slightly by fetal calf serum. Biochemical characterization of the PAPK-A-initiated pathway revealed that this novel kinase does not affect MAP kinase activity but can stimulate both c-Jun N-terminal kinase 1 (JNK1) and ERK6/p38 gamma. The kinase activity of PAPK appears to be required for the activation of ERK6/p38 gamma but not JNK1. When an inducible construct of PAPK-A was expressed in stably transfected NIH3T3 cells, the cells exhibited distinct cytoskeletal changes and became resistant to apoptotic cell death induced by serum withdrawal, effects of PAPK that require its kinase activity. These data suggest that PAPK is a new member of the Ste20/germinal center kinase family that modulates cytoskeletal organization and cell survival. PMID- 12574164 TI - Structural characterization of a human cytosolic NMN/NaMN adenylyltransferase and implication in human NAD biosynthesis. AB - Pyridine dinucleotides (NAD and NADP) are ubiquitous cofactors involved in hundreds of redox reactions essential for the energy transduction and metabolism in all living cells. In addition, NAD also serves as a substrate for ADP ribosylation of a number of nuclear proteins, for silent information regulator 2 (Sir2)-like histone deacetylase that is involved in gene silencing regulation, and for cyclic ADP ribose (cADPR)-dependent Ca(2+) signaling. Pyridine nucleotide adenylyltransferase (PNAT) is an indispensable central enzyme in the NAD biosynthesis pathways catalyzing the condensation of pyridine mononucleotide (NMN or NaMN) with the AMP moiety of ATP to form NAD (or NaAD). Here we report the identification and structural characterization of a novel human PNAT (hsPNAT-3) that is located in the cytoplasm and mitochondria. Its subcellular localization and tissue distribution are distinct from the previously identified human nuclear PNAT-1 and PNAT-2. Detailed structural analysis of PNAT-3 in its apo form and in complex with its substrate(s) or product revealed the catalytic mechanism of the enzyme. The characterization of the cytosolic human PNAT-3 provided compelling evidence that the final steps of NAD biosynthesis pathways may exist in mammalian cytoplasm and mitochondria, potentially contributing to their NAD/NADP pool. PMID- 12574165 TI - Characterization of a cAMP-stimulated cAMP phosphodiesterase in Dictyostelium discoideum. AB - A cyclic nucleotide phosphodiesterase, PdeE, that harbors two cyclic nucleotide binding motifs and a binuclear Zn(2+)-binding domain was characterized in Dictyostelium. In other eukaryotes, the Dictyostelium domain shows greatest homology to the 73-kDa subunit of the pre-mRNA cleavage and polyadenylation specificity factor. The Dictyostelium PdeE gene is expressed at its highest levels during aggregation, and its disruption causes the loss of a cAMP phosphodiesterase activity. The pdeE null mutants show a normal cAMP-induced cGMP response and a 1.5-fold increase of cAMP-induced cAMP relay. Overexpression of a PdeE-yellow fluorescent protein (YFP) fusion construct causes inhibition of aggregation and loss of the cAMP relay response, but the cells can aggregate in synergy with wild-type cells. The PdeE-YFP fusion protein was partially purified by immunoprecipitation and biochemically characterized. PdeE and its Dictyostelium ortholog, PdeD, are both maximally active at pH 7.0. Both enzymes require bivalent cations for activity. The common cofactors Zn(2+) and Mg(2+) activated PdeE and PdeD maximally at 10 mm, whereas Mn(2+) activated the enzymes to 4-fold higher levels, with half-maximal activation between 10 and 100 microm. PdeE is an allosteric enzyme, which is approximately 4-fold activated by cAMP, with half-maximal activation occurring at about 10 microm and an apparent K(m) of approximately 1 mm. cGMP is degraded at a 6-fold lower rate than cAMP. Neither cGMP nor 8-Br-cAMP are efficient activators of PdeE activity. PMID- 12574166 TI - RhoA and ROCK promote migration by limiting membrane protrusions. AB - Previously, we and others have shown that RhoA and ROCK signaling are required for negatively regulating integrin-mediated adhesion and for tail retraction of migrating leukocytes. This study continues our investigation into the molecular mechanisms underlying RhoA/ROCK-regulated integrin adhesion. We show that inhibition of ROCK up-regulates integrin-mediated adhesion, which is accompanied by both increased phosphotyrosine signaling through Pyk-2 and paxillin and inappropriate membrane protrusions. We provide evidence that inhibition of ROCK induces integrin adhesion by promoting remodeling of the actin cytoskeleton. Furthermore, we find that ROCK regulates membrane activity through a pathway involving cofilin. Inhibition of RhoA signaling allows the formation of multiple competing lamellipodia that disrupt productive migration of monocytes. Together, our results show that RhoA/ROCK signaling promotes migration by restricting integrin activity and membrane protrusions to the leading edge. PMID- 12574167 TI - ErbB2 degradation mediated by the co-chaperone protein CHIP. AB - ErbB2 overexpression contributes to the evolution of a substantial group of human cancers and signifies a poor clinical prognosis. Thus, down-regulation of ErbB2 signaling has emerged as a new anti-cancer strategy. Ubiquitinylation, mediated by the Cbl family of ubiquitin ligases, has emerged as a physiological mechanism of ErbB receptor down-regulation, and this mechanism appears to contribute to ErbB2 down-regulation induced by therapeutic anti-ErbB2 antibodies. Hsp90 inhibitory ansamycin antibiotics such as geldanamycin (GA) induce rapid ubiquitinylation and down-regulation of ErbB2. However, the ubiquitin ligase(s) involved has not been identified. Here, we show that ErbB2 serves as an in vitro substrate for the Hsp70/Hsp90-associated U-box ubiquitin ligase CHIP. Overexpression of wild type CHIP, but not its U-box mutant H260Q, induced ubiquitinylation and reduction in both cell surface and total levels of ectopically expressed or endogenous ErbB2 in vivo, and this effect was additive with that of 17-allylamino-geldanamycin (17-AAG). The CHIP U-box mutant H260Q reduced 17-AAG-induced ErbB2 ubiquitinylation. Wild type ErbB2 and a mutant incapable of association with Cbl (ErbB2 Y1112F) were equally sensitive to CHIP and 17-AAG, implying that Cbl does not play a major role in geldanamycin-induced ErbB2 down-regulation. Both endogenous and ectopically expressed CHIP and ErbB2 coimmunoprecipitated with each other, and this association was enhanced by 17 AAG. Notably, CHIP H260Q induced a dramatic elevation of ErbB2 association with Hsp70 and prevented the 17-AAG-induced dissociation of Hsp90. Our results demonstrate that ErbB2 is a target of CHIP ubiquitin ligase activity and suggest a role for CHIP E3 activity in controlling both the association of Hsp70/Hsp90 chaperones with ErbB2 and the down-regulation of ErbB2 induced by inhibitors of Hsp90. PMID- 12574168 TI - A hybrid IRF9-STAT2 protein recapitulates interferon-stimulated gene expression and antiviral response. AB - Type I interferon (IFN) signaling induces the heterotrimeric transcription complex, IFN-stimulated gene factor (ISGF) 3, which contains STAT1, STAT2, and the DNA binding subunit, interferon regulatory factor (IRF) 9. Because IRF9 is targeted to the nucleus in the absence of IFN stimulation, the potential of IRF9 protein for gene regulation was examined using a GAL4 DNA binding domain fusion system. GAL4-IRF9 was transcriptionally active in reporter gene assays but not in the absence of cellular STAT1 and STAT2. However, the inert IRF9 protein was readily converted to a constitutively active ISGF3-like activator by fusion with the C-terminal transcriptional activation domain of STAT2 or the acidic activation domain of herpesvirus VP16. The IRF9 hybrids are targeted to endogenous ISGF3 target loci and can activate their transcription. Moreover, expression of the IRF9-STAT2 fusion can recapitulate the type I IFN biological response, producing a cellular antiviral state that protects cells from virus induced cytopathic effects and inhibits virus replication. The antiviral state generated by regulated IRF9-STAT2 hybrid protein expression is independent of autocrine IFN signaling and inhibits both RNA and DNA viruses. PMID- 12574169 TI - Mouse GGN1 and GGN3, two germ cell-specific proteins from the single gene Ggn, interact with mouse POG and play a role in spermatogenesis. AB - The germ cell-deficient (gcd) mutation is a recessive transgenic insertional mutation leading to a deficiency of primordial germ cells (PGCs). We have recently shown that the gene underlying this mutation is Pog, which is necessary for normal proliferation of PGCs. Here we show that Pog is also involved in spermatogenesis in that meiosis is impaired in Pog-deficient mice. Yeast two hybrid screening revealed that POG interacted with GGN1 and GGN3, two proteins formed by alternate splicing of the same gene, gametogenetin (Ggn). Ggn had more than 10 different splice variants giving rise to three proteins, GGN1, GGN2, and GGN3. The three proteins had different subcellular localizations, with GGN1, GGN2, and GGN3 localized along the nuclear membrane, in the cytoplasm, and in the nucleus/nucleoli respectively. The expression of Ggn was confined to late pachytene spermatocytes and round spermatids, a time window concomitant with the occurrence of meiosis. Mouse Ggn and Pog were both expressed in primary spermatocytes. Co-expression of POG with GGN1 or GGN3 in HeLa cells changed the localization of POG to the perinuclear localization or the nucleoli, respectively. Our data showed that in addition to functioning in proliferation of primordial germ cells, POG also functioned in spermatogenesis. Two spatial and temporal regulated proteins, GGN1 and GGN3, interacted with POG, regulated the localization of POG, and played a role in spermatogenesis. PMID- 12574170 TI - Salmonella typhimurium SifA effector protein requires its membrane-anchoring C terminal hexapeptide for its biological function. AB - SifA is a Salmonella typhimurium effector protein that is translocated across the membrane of the Salmonella-containing vacuole by the Salmonella pathogenicity island 2-encoded type III secretion system. SifA is necessary for the formation of Salmonella-induced filaments and for the maintenance of the vacuolar membrane enclosing the pathogen. We have investigated the role of the C-terminal hexapeptide of SifA as a potential site for membrane anchoring. An S. typhimurium strain carrying a deletion of the sequence encoding this hexapeptide (sifA Delta 6) was found to be attenuated for systemic virulence in mice. In mouse macrophages, sifA Delta 6 mutant bacteria displayed a reduced association with vacuolar markers, similar to that of sifA null mutant bacteria, and exhibited a dramatic replication defect. Expression of SifA in epithelial cells results in the mobilization of lysosomal glycoproteins in large vesicular structures and Sif like tubules. This process requires the presence of the C-terminal hexapeptide domain of SifA. Ectopic expression of truncated or mutated versions of SifA affecting the C-terminal hexapeptide revealed a strong correlation between the membrane binding capability and the biological activity of the protein. Finally, the eleven C-terminal residues of SifA are shown to be sufficient to target the Aequorea green fluorescent protein to membranes. Altogether, our results indicate that membrane anchoring of SifA requires its C-terminal hexapeptide domain, which is important for the biological function of this bacterial effector. PMID- 12574172 TI - Cardiovascular endocrinology 1: aldosterone function in diabetes mellitus: effects on cardiovascular and renal disease. PMID- 12574174 TI - Editorial: sulfonylureas differ in effects on ischemic preconditioning--is it time to retire glyburide? PMID- 12574175 TI - Impairment of myocardial protection in type 2 diabetic patients. AB - Diabetic patients are more prone to develop postinfarction complications. It remained unclear whether diabetes mellitus- or sulfonylureas-associated changes of ATP-sensitive potassium (K(ATP)) channels, an integral player in ischemic preconditioning, are responsible for the increased mortality. The purpose of this study was to determine the impact of diabetes mellitus per se and different sulfonylurea administration on cardioprotective effects in diabetic patients undergoing coronary angioplasty. Myocardial ischemia after coronary angioplasty was evaluated in 20 nondiabetic and 23 diabetic patients chronically taking either glibenclamide or glimepiride. Nondiabetic patients treated with glimepiride significantly lowered the ischemic burden assessed by an ST-segment shift, chest pain score, and myocardial lactate extraction ratios compared with the glibenclamide-treated patients, implying that acute administration of glimepiride did not abolish cardioprotection. In the diabetic glibenclamide treated group, the reduction in the ST-segment shift afforded by nicorandil in the first inflation (-58% vs. the first inflation in the glibenclamide group alone) was similar to that afforded by preconditioning (-59% during the second vs. the first inflation). In glimepiride-treated groups, the magnitude of attenuated lactate production was less in diabetes than that in nondiabetes at the second inflation, suggesting that diabetes mellitus per se plays a role in determining lactate production. Our results show that both diabetes mellitus and sulfonylureas can act in synergism to inhibit activation of K(ATP) channels in patients undergoing coronary angioplasty. The degree of inhibition assessed by metabolic and electrocardiographic parameters is less severe during treatment with glimepiride than with glibenclamide. Restitution of a preconditioning response in glimepiride-treated patients may be the potential beneficial mechanism. PMID- 12574176 TI - Editorial: greater risk, greater benefit--true or false? PMID- 12574177 TI - Risedronate prevents new vertebral fractures in postmenopausal women at high risk. AB - Independent risk factors for fracture include advanced age, preexisting fractures, and low bone mineral density. Rised-ronate has been shown in several large trials to be safe and effective for patients with osteoporosis, but its effects in populations at high risk are not well characterized. To determine the effect of risedronate on vertebral fracture in high-risk subjects, we pooled data from two randomized, double-blind studies [Vertebral Efficacy with Risedronate Therapy (VERT) Multinational (VERT-MN) and VERT-North America (VERT-NA)] in 3684 postmenopausal osteoporotic women treated with placebo or risedronate 2.5 or 5 mg/d and analyzed fracture risk in subgroups of subjects at high risk for fracture due to greater age or more prevalent fractures (vs. median for overall study population), or lower bone mineral density (T-score, -2.5 or less). Fractures were diagnosed by quantitative and semiquantitative assessment of radiographs at baseline and 1 yr. In the overall population, treatment for 1 yr with risedronate 5 mg/d reduced the risk of new vertebral fractures by 62% vs. control (relative risk, 0.38; 95% confidence interval, 0.25, 0.56; P < 0.001) and of multiple new vertebral fractures by 90% vs. control (relative risk, 0.10; 95% confidence interval, 0.04, 0.26; P < 0.001). Consistent risk reductions were observed at 1 yr in the risedronate-treated high-risk subgroups. Significant reduction in fracture risk after 1 yr is an important benefit in patients at high risk for fracture because, without treatment, these patients are likely to sustain new fractures in the near term. PMID- 12574178 TI - Editorial: biochemical diagnosis of pheochromocytoma--is it time to switch to plasma-free metanephrines? PMID- 12574179 TI - A comparison of biochemical tests for pheochromocytoma: measurement of fractionated plasma metanephrines compared with the combination of 24-hour urinary metanephrines and catecholamines. AB - We compared the diagnostic efficacy of fractionated plasma metanephrine measurements to measurements of 24-h urinary total metanephrines and catecholamines in outpatients tested for pheochromocytoma at Mayo Clinic Rochester from January 1, 1999, until November 27, 2000. Catecholaminesecreting tumors were histologically proven. The sensitivity of fractionated plasma metanephrines was 97% (30 of 31 patients), compared with a sensitivity of 90% (28 of 31) for urinary total metanephrines and catecholamines (P = 0.63). The specificity of fractionated plasma metanephrines was 85% (221 of 261), compared with 98% (257 of 261; P < 0.001) for urinary measurements. The likelihood ratios for positive tests were 6.3 (95% confidence interval, 4.7 to 8.5) for fractionated plasma metanephrines and 58.9 (95% confidence interval, 22.1 to 156.9) for urinary total metanephrines and catecholamines. An adrenal pheochromocytoma was missed by urinary testing in two patients with familial syndromes and one asymptomatic patient with an incidentally discovered adrenal mass. An extra-adrenal paraganglioma was missed by plasma testing in one patient. In conclusion, measurements of 24-h urinary total metanephrines and catecholamines yield fewer false-positive results, an attribute preferred for testing low-risk patients, but fractionated plasma metanephrine measurements may be preferred in high-risk patients with familial endocrine syndromes. PMID- 12574180 TI - Editorial: hormonal male contraception--lessons from the East when the Western market fails. PMID- 12574181 TI - A multicenter contraceptive efficacy study of injectable testosterone undecanoate in healthy Chinese men. AB - This report describes a Phase II, multicenter, contraceptive efficacy clinical trial using monthly injections of testosterone undecanoate (TU) alone at a dose of 500 mg in healthy Chinese men. Three hundred eight healthy men were recruited in six centers distributed throughout China. Volunteers underwent a control period with no treatment, then a 12-month treatment period including a 6-month suppression phase followed by a 6-month efficacy phase and a 12-month recovery period. During the suppression phase, an initial loading dose of 1000 mg TU, followed by 500 TU at monthly intervals were given until azoospermia or severe oligozoospermia was achieved, up to a maximum of six injections. During the efficacy phase, 500 mg TU were administered at monthly intervals for 6 months. Nine of 308 men did not achieve azoospermia or severe oligozoospermia (<3 x 10(6)/ml) within the 6-month suppression phase. This gave a methodological failure rate of 2.9/100 couple years (95% confidence interval of 1.0-4.8/100 couple years). Two hundred ninety-six men entered the efficacy phase. The continuation rate during the efficacy phase was 95/100 couple years. There were no pregnancies caused by men who achieved azoospermia or severe oligozoospermia. Reappearance of sperm occurred in six men during the efficacy phase, and one pregnancy was attributed to sperm rebound. This gave a secondary failure rate of 2.3/100 couple years (95% confidence interval of 0.5-4.2/100 couple years). Thus, the total failure rate was 5.2%, and total efficacy was 94.8%. Spermatogenesis in all subjects returned to the normal reference range within the recovery period. The mean serum testosterone concentration increased 131%, and the mean serum LH and FSH concentrations decreased 72% and 70%, respectively, after TU injections during the treatment period. The mean level of serum high density lipoprotein cholesterol decreased (14%), and the mean hematocrit increased 6% compared with baseline. No serious adverse events and no significant changes in serum chemistry occurred during the study. The results showed that monthly TU injection at a dose of 500 mg after an initial loading dose of 1000 mg can effectively, safely, and reversibly suppress spermatogenesis in healthy Chinese men without serious adverse effects. PMID- 12574182 TI - Short-term, high-dose parathyroid hormone-related protein as a skeletal anabolic agent for the treatment of postmenopausal osteoporosis. AB - PTH-related protein (PTHrP) is homologous with PTH. PTH, an effective anabolic agent for treating osteoporosis, has been shown to stimulate both bone resorption by osteoclasts and bone formation by osteoblasts. We examined whether PTHrP might share anabolic properties in osteoporosis. A 3-month double-blind, prospective, placebo-controlled, randomized clinical trial was performed in 16 healthy postmenopausal women with osteoporosis. All received calcium and vitamin D, and all continued their prior hormone replacement therapy. One group also received daily sc PTHrP (6.56 microg/kg x d, or approximately 400 microg/d), and the other group received placebo injections. The PTHrP group displayed a 4.7% increase in lumbar spine bone mineral density (BMD) and also demonstrated an increase in osteoblastic bone formation, as assessed using serum osteocalcin measurements. In contrast, there was no increase in bone-specific alkaline phosphatase and collagen-1 propeptide or either of two markers of osteoclastic bone resorption, N telopeptide, or deoxypyridinoline. One subject in the placebo group withdrew from the study, but there were no significant adverse events in the PTHrP group. PTHrP administered sc in high doses for only 3 months appears to be a potent anabolic agent, producing a 4.7% increase in lumbar spine BMD. This compares very favorably to available antiresorptive drugs for osteoporosis and is similar to the increases in BMD at this early time point reported for PTH. Despite the high doses, PTHrP was well tolerated. Larger clinical trials are required to confirm these results and fully assess the anabolic potential of PTHrP in osteoporosis. PMID- 12574183 TI - Troglitazone therapy improves endothelial function to near normal levels in women with polycystic ovary syndrome. AB - Obese women with polycystic ovary syndrome (PCOS) exhibit impaired endothelial function, which is strongly and directly correlated with both testosterone levels and insulin resistance. Endothelial dysfunction is considered a potent risk factor for macrovascular disease. Because troglitazone (Tgz) improves both hormonal profiles and insulin sensitivity, we tested whether Tgz treatment ameliorates endothelial function in these patients. We studied leg blood flow (LBF) responses to graded intrafemoral artery infusion of the endothelium dependent vasodilator methacholine chloride (MCh) and to a 4-h hyperinsulinemic euglycemic clamp (120 mU/m(2) x min) in 10 PCOS, before and after 3 months treatment with Tgz (600 mg/d). A group of 13 obese women (OBW) matched for age, weight, body fat (>40% in both groups), blood pressure, and total cholesterol served as controls. PCOS patients exhibited elevated free testosterone (fT) and triglycerides (TG) and lower high density lipoprotein cholesterol levels compared with OBW [14.0 +/- 1.0 vs. 3.7 +/- 0.6 pmol/liter (P < 0.0001), 1.60 +/- 0.28 vs. 0.94 +/- 0.09 mmol/liter (P < 0.02), and 0.91 +/- 0.04 vs. 1.1 +/- 0.04 mmol/liter (P < 0.005), respectively]. Tgz treatment reduced fT levels, but did not improve the TG and high density lipoprotein profile [to 9.7 +/- 2.8 pmol/liter (P < 0.007), 1.49 +/- 0.34 mmol/liter (P = NS), and 0.93 +/- 0.07 mmol/liter (P = NS), respectively]. Basal LBF was unchanged after Tgz. In PCOS compared with OBW, insulin stimulated glucose disposal (52.7 +/- 6.6 vs. 85.5 +/- 4.4 micromol/kg fat-free mass x min; P < 0.0005) and vasodilation (increase in LBF, 22 +/- 14% vs. 59 +/- 15%; P < 0.05) were significantly improved after Tgz treatment to 68.8 +/- 7.2 micromol/kg fat-free mass x min (P < 0.0001) and 101 +/ 48% (P < 0.03), respectively. The increase in LBF in response to MCh in PCOS was markedly more pronounced after treatment (P < 0.01, by ANOVA) and was similar to that observed in OBW. Before Tgz treatment, maximal LBF increments in response to MCh were 130 +/- 25% and 233 +/- 29% in PCOS and OBW, respectively (P < 0.01). After Tgz treatment, PCOS values improved, achieving increments similar to those in OBW (245 +/- 45%; P < 0.04). Tgz treatment in PCOS improves both hormonal and metabolic features. These modifications are associated with improvement of endothelial function, suggesting that Tgz could be a useful tool to reduce the risk of macrovascular disease in women with PCOS and perhaps in other insulin resistant syndromes. PMID- 12574184 TI - Oral alendronate increases bone mineral density in postmenopausal women with primary hyperparathyroidism. AB - The effect of biphosphonate therapy on bone mineral density (BMD) in patients with primary hyperparathyroidism (PHP) is unknown. Forty postmenopausal women (mean age, 70 yr) with PHP were randomized to receive alendronate 10 mg/d or placebo for 48 wk, followed by treatment withdrawal for 24 wk. The mean (+/-SD) changes in BMD at femoral neck (+4.17 +/- 6.01% vs. -0.25 +/- 3.3%; P = 0.011) and lumbar spine (+3.79 +/- 4.04% vs. 0.19 +/- 2.80%; P = 0.016) were significantly higher with alendronate at 48 wk. Serum calcium was reduced with alendronate but not placebo (-0.09 vs. +0.01 mmol/liter; P = 0.018). Serum bone specific alkaline phosphatase activity was lower with alendronate from 12 wk onward and increased 24 wk after treatment withdrawal (21.1 +/- 12.8 to 7.3 +/- 4.9 IU/liter at 48 wk, and 15.0 +/- 14.8 IU/liter 24 wk after withdrawal; P = 0.002 for trend). Osteocalcin concentration decreased at 48 wk and increased 24 wk after alendronate withdrawal (P = 0.019 for trend of change over time) but not with placebo. Urinary N-telopeptide/creatinine ratio decreased with alendronate at 48 wk and increased 24 wk after treatment withdrawal (P = 0.008 for trend). N telopeptide/creatinine ratio did not change with placebo. Alendronate improves BMD and reduces bone turnover markers in postmenopausal women with PHP. PMID- 12574185 TI - Differential effects of oral and transdermal estradiol treatment on circulating estradiol fatty acid ester concentrations in postmenopausal women. AB - Estradiol fatty acid esters are potent lipophilic estrogens with antioxidant properties, transported by lipoproteins in blood. We investigated effects of oral and transdermal estradiol replacement therapy on concentrations of estradiol fatty acid esters in serum in postmenopausal women in a double-blind, randomized fashion. The first group (n = 9) received oral (2 mg/d); the second (n = 10), transdermal estradiol (patch delivering 50 microg/d); and the third group (n = 7), placebo treatment for 12 wk. After extraction of serum and separation of esterified estradiol from nonesterified estradiol, the concentration of saponified estradiol esters was measured by time-resolved fluoroimmunoassay. In the oral estradiol group, the median serum estradiol fatty acid ester concentration rose by 27%, from 77 to 98 pM (P = 0.028) but remained unchanged in the transdermal estradiol and placebo groups. The median concentrations of serum nonprotein-bound estradiol increased similarly in the oral and transdermal estradiol groups. The change in serum estradiol ester concentrations during treatment, but not that of nonesterified estradiol, correlated positively with enhanced forearm blood flow responses in vivo. These data raise the possibility that an increase in serum estradiol fatty acid esters may contribute to beneficial effects of oral estradiol treatment, compared with an equipotent dose of transdermal estradiol. PMID- 12574186 TI - Overexpression of type 2 iodothyronine deiodinase in follicular carcinoma as a cause of low circulating free thyroxine levels. AB - Thyroid function is normally undisturbed in patients with thyroid carcinoma. We have identified three patients with large or widely metastatic follicular thyroid carcinoma who had a persistently increased ratio of serum T(3) to T(4) in the absence of autonomous production of T(3) by the tumor. To investigate the possibility of tumor-mediated T(4) to T(3) conversion, we assayed types 1 and 2 iodothyronine selenodeiodinase (D1 and D2) activity in a 965-g follicular thyroid carcinoma resected from one of these patients. The V(max) for D2 was 8-fold higher than in normal human thyroid tissue. Resection of this tumor, leaving the left thyroid lobe intact, normalized the serum T(3) to T(4) ratio. In two other patients, treatment with sufficient levothyroxine to suppress TSH was associated with a high normal T(3) and a subnormal free T(4) index. In one, concomitant administration of the D1 inhibitors, propylthiouracil and propranolol, did not decrease the elevated serum T(3) to T(4) ratio. These data illustrate that increased T(4) to T(3) conversion in follicular thyroid carcinomas, probably by D2, can cause a significant perturbation in peripheral thyroid hormone concentrations. PMID- 12574187 TI - Relationship of leptin to bone mineralization in children and adolescents. AB - Serum leptin concentrations and bone mass are concordant in several respects. Obesity is associated with increased serum leptin concentrations and bone mineral, whereas undernutrition reduces serum leptin concentrations and bone mineral. Furthermore, both bone mineral and serum leptin concentrations increase at the initiation of puberty. However, there is a lack of empirical evidence of an independent association of serum leptin concentrations and bone mineral in youth. Thus, we used hierarchical regression to determine whether serum leptin concentrations were related to bone mineral in boys (n = 28) and girls (n = 31). Bone mineral content, density, and apparent density of the total body and body regions were measured by dual-energy x-ray absorptiometry and statistically adjusted for chronological age, fat mass, bone-free fat-free mass, and serum IGF I and estradiol concentrations. Sequential addition of serum log((10)) leptin concentrations to the block of body size variables and the block of hormone variables did not increase R(2) for any of the total or regional bone mineral content, bone mineral density, and bone mineral apparent density variables. We conclude that serum leptin concentrations do not add to the prediction of bone mineral in youth after accounting for age, fat mass, bone-free fat-free mass, and serum IGF-I and estradiol concentrations. PMID- 12574188 TI - Autosomal dominant hypocalcemia: a novel activating mutation (E604K) in the cysteine-rich domain of the calcium-sensing receptor. AB - We report a novel activating mutation (E604K) of the calcium-sensing receptor in a family with autosomal dominant hypocalcemia. Whereas all affected individuals exhibited marked hypocalcemia, some cases with untreated hypocalcemia exhibited seizures in infancy, whereas others were largely asymptomatic from birth into adulthood. The missense mutation E604K (G2182A; GenBank accession no. U20759), which affects an amino acid residue in the C terminus of the cysteine-rich domain of the extracellular head, cosegregated with hypocalcemia in all seven individuals for whom DNA was available. Two unaffected, normocalcemic members of the family did not exhibit the mutation. The molecular impact of the mutation on two key components of the signaling response was assessed in HEK-293 cells transiently transfected with cDNA corresponding to either the wild-type calcium sensing receptor or the E604K mutation derived by site-directed mutagenesis. There was a significant leftward shift in the concentration response curves for the effects of extracellular Ca(2+) on both intracellular Ca(2+) mobilization (determined by aequorin luminescence) and MAPK activity (determined by luciferase expression). The C terminus of the cysteine-rich domain of the extracellular head may normally act to suppress receptor activity in the presence of low extracellular Ca(2+) concentrations. PMID- 12574189 TI - Risk factors for the development of obesity in children surviving brain tumors. AB - Hypothalamic obesity, a syndrome of intractable weight gain due to hypothalamic damage, is an uncommon but devastating complication for children surviving brain tumors. We undertook a retrospective evaluation of the body mass index (BMI) curves for the St. Jude Children's Research Hospital brain tumor population diagnosed between 1965 and 1995 after completion of therapy to determine risk factors for the development of obesity. Inclusion criteria were: diagnosis less than 14 yr of age, no spinal cord involvement, ambulatory, no supraphysiologic hydrocortisone therapy (>12 mg/m(2) x d), treatment and follow-up at St. Jude Children's Research Hospital, and disease-free survival greater than 5 yr (n = 148). Risk factors examined were age at diagnosis, tumor location, histology, extent of surgery, hydrocephalus requiring ventriculoperitoneal shunting, initial high-dose glucocorticoids, cranial radiation therapy, radiation dosimetry to the hypothalamus, intrathecal chemotherapy, and presence of endocrinopathy. Analyses were performed both between groups within a risk factor and against BMI changes for age in normal children older than 5.5 yr (the age of adiposity rebound). Risk factors were: age at diagnosis (P = 0.04), radiation dosimetry to the hypothalamus (51-72 Gy, P = 0.002 even after hypothalamic and thalamic tumor exclusion), and presence of any endocrinopathy (P = 0.03). In addition, risk factors when compared with BMI slope for the general American pediatric population included: tumor location (hypothalamic, P = 0.001), tumor histology (craniopharyngioma, P = 0.009; pilocytic astrocytoma, P = 0.043; medulloblastoma, P = 0.039); and extent of surgery (biopsy, P = 0.03; subtotal resection, P = 0.018). These results verify hypothalamic damage, either due to tumor, surgery, or radiation, as the primary cause of obesity in survivors of childhood brain tumors. In particular, hypothalamic radiation doses of more than 51 Gy are permissive. These results reiterate the importance of the hypothalamus in energy balance, provide risk assessment criteria for preventative measures before the development of obesity in at-risk patients, and suggest therapeutic strategies to reduce the future development of obesity. PMID- 12574190 TI - Hyperthyrotropinemia during iodide administration in normal children and in children born with neonatal transient hypothyroidism. AB - The aim of the present study was to examine the effects of chronic iodide administration in pharmacological doses on thyroid function in children with a history of transient congenital hypothyroidism (TCH). We hypothesized that such children may carry a previously undisclosed intrinsic intrathyroidal defect, rendering them susceptible to TCH. We administered for this 60-65 mg iodide daily for 60 d in 13 individuals with TCH (group A), 8 of their siblings (group B), 8 healthy controls (group C), and 11 normal adults (group D). Thyroid function was evaluated by measuring serum T(3), T(4), free T(3), free T(4), TSH, and thyroglobulin concentrations and autoantibodies against thyroid peroxidase and thyroglobulin at baseline at 15, 30, and 60 d during iodide administration, and 2 months after iodide withdrawal. Hyperthyrotropinemia greater than 4.2 mU/liter but not higher than 10 mU/liter with normal thyroid hormone concentrations was observed in one of the TCH group and in two of the group B siblings. During iodide administration, hyperthyrotropinemia was observed in 8 of 13 (62%) adolescents in group A, 4 of 7 (57%) in group B, and 6 of 8 (75%) in group C. None of the 11 adults (group D) developed hyperthyrotropinemia during iodide administration. Serum T(4) and free T(4) concentrations were decreased in all groups when compared with baseline values. The magnitude of the decrease of serum T(4) was identical in all groups (0.7-0.8 microg/dl). Thyroid enlargement was observed in all subjects and was more pronounced in children. There were no cases of subclinical and/or overt hyperthyroidism. After iodine withdrawal, serum TSH decreased in all groups and returned to baseline levels, as well as the thyroid volume. In conclusion, the hypothalamic-pituitary-thyroid axis of adolescents with TCH responds to pharmacological doses of iodide similarly to that observed in normal children. The hyperthyrotropinemia observed in the adolescents exposed to iodides may reflect incipient transient hypothyroidism or simply a brisk TSH response to a small serum T(4) decrease. Whatever the mechanism, chronic use of excessive quantities of iodide should be avoided until the end of puberty. PMID- 12574191 TI - Prenatal and postnatal prevalence of Klinefelter syndrome: a national registry study. AB - The objective of this study was to describe the prevalence of Klinefelter syndrome (KS) prenatally and postnatally in Denmark and determine the influence of maternal age. All chromosomal examinations in Denmark are registered in the Danish Cytogenetic Central Registry. Individuals with KS diagnosed prenatally or postnatally were extracted from the registry with information about age at the time of diagnosis and mother's age. In the period 1970-2000, 76,526 prenatal examinations on male fetuses resulted in the diagnosis of 163 fetuses with KS karyotype, corresponding to a prevalence of 213 per 100,000 male fetuses. Standardization according to maternal age resulted in a prevalence of 153 per 100,000 males. Postnatally, 696 males of 2,480,858 live born were diagnosed with KS, corresponding to a prevalence among adult men of approximately 40 per 100,000. Less than 10% of the expected number was diagnosed before puberty. Advanced maternal age had a significant impact on the prevalence. KS is severely underdiagnosed in Denmark. Only approximately one fourth of adult males with KS are diagnosed. There is a marked delay in diagnosis of the syndrome. A delay in treatment with testosterone may lead to decreased muscle and bone mass with subsequent risk of osteoporosis. PMID- 12574192 TI - Hypoadiponectinemia is associated with insulin resistance, hypertriglyceridemia, and fat redistribution in human immunodeficiency virus-infected patients treated with highly active antiretroviral therapy. AB - A lipodystrophic syndrome and metabolic abnormalities have been observed in HIV infected patients treated with highly active antiretroviral therapy (HAART). A murine model of lipodystrophy is associated with decreased levels of adiponectin, an adipocyte-secreted protein, the administration of which improves the metabolic syndrome in these mice. To investigate the association of adiponectin with metabolic changes in human lipodystrophy, we conducted a cross-sectional study of 112 HIV-infected patients treated with HAART. Mean adiponectin levels were higher in patients with no fat redistribution (FR) vs. FR (4.8 +/- 5.0 vs. 2.2 +/- 2.7 microg/ml, P < 0.01), but no significant differences in adiponectin levels were observed between FR subgroups. The difference in adiponectin levels between subjects with and without FR remained significant after adjusting for age, gender, leptin, HIV medication use, and CD4 count using logistic regression (odds ratio, 0.54, P = 0.008). Adiponectin was significantly correlated with triglycerides (r = -0.40), abdominal visceral fat (r = -0.35), extremity fat (r = 0.37), insulin resistance (HOMA-IR) (r = -0.28), nucleoside reverse transcriptase inhibitor (NRTI) use (r = -0.32), and high-density lipoprotein (HDL) (r = 0.41) using bivariate analysis (all P < 0.01). The association with HDL weakened but remained significant on multivariate analysis (standard beta = 0.29, P = 0.01). However, the association of adiponectin with HOMA-IR became nonsignificant after adjusting for NRTI use (standard beta = -0.15, P = 0.12), suggesting that changes in adiponectin levels may underlie the effect of NRTI use on insulin resistance. The associations of adiponectin with triglycerides and HOMA-IR were also slightly weakened after adjusting for visceral and extremity fat, indicating that adiponectin may, in part, mediate the effect of FR on triglycerides and insulin resistance. This study indicates that adiponectin is inversely correlated with abdominal visceral fat mass, serum triglycerides, and insulin resistance and is directly correlated with HDL and extremity fat in a sample of HIV-infected patients treated with HAART. The results also indicate that NRTI use may worsen insulin resistance by decreasing adiponectin levels. Thus, adiponectin replacement may be a potential treatment option to ameliorate the metabolic changes observed in this patient population. PMID- 12574193 TI - Localization of medullary thyroid carcinoma metastasis in a multiple endocrine neoplasia type 2A patient by 6-[18F]-fluorodopamine positron emission tomography. AB - 6-[(18)F]fluorodopamine, a substrate for the norepinephrine transporter, has been used as a tumor-seeking tracer in positron emission tomography (PET) to localize pheochromocytomas and other chromaffin tumors. Here, we report the case of a 42 yr-old woman with multiple endocrine neoplasia type 2A, in whom biopsy-proven recurrent medullary thyroid cancer (MTC) was detected by 6-[(18)F]fluorodopamine PET scanning. The patient had previously undergone bilateral adrenalectomy for pheochromocytoma, total thyroidectomy, and extirpation of a parapharyngeal MTC metastatic deposit. An increase in plasma calcitonin 5 yr after her initial presentation was further investigated, leading to the discovery of a mass in the left parapharyngeal space. Levels of serum and urine catecholamines and metanephrines were normal. To exclude a hormonally silent pheochromocytoma metastasis, 6-[(18)F]fluorodopamine PET was performed. The study showed a focus of radionuclide accumulation corresponding to the parapharyngeal mass. After resection of the latter, pathology confirmed metastatic MTC. To our knowledge, this is the first case of metastatic, histologically proven MTC, which was unequivocally detected by 6-[(18)F]fluorodopamine PET scanning. Because norepinephrine transporter systems have been previously found in MTC, it is conceivable that 6-[(18)F]fluorodopamine PET scanning can be used for the diagnostic localization of this tumor and its metastatic deposits because total and early resection is beneficial to the outcome of the patient. PMID- 12574194 TI - Difference in bone mass between black and white American children: attributable to body build, sex hormone levels, or bone turnover? AB - A cross-sectional study of 232 healthy children, with about equal numbers of boys and girls and blacks and whites, aged 4 to 16 yr, was conducted to investigate the racial differences in bone mineral. Bone mineral content (BMC) by dual x-ray absorptiometry was found to be similar between blacks and whites at the spine after controlling for age and Tanner stage. However, total body BMC was higher in blacks, compared with whites of the same age and Tanner stage. Height and weight alone reduced the racial difference in BMC from 152 g to 66 g in girls and from 163 g to 105 g in boys, in whom the difference was further reduced to 66 g after accounting for lean and fat body mass and subscapular skinfold. The only significant sex hormone was androstenedione, which explained another 4-5 g of the racial difference in total body BMC for both boys and girls. Among the biochemical variables, only 25OH vitamin D reduced the residual racial difference in total body BMC to 39 g in girls, whereas serum PTH, urine free deoxypyridinoline ratio, and 1,25(OH)(2) vitamin D reduced the residual difference to 25 g in boys. The residual racial differences in bone mass were not statistically significant. PMID- 12574195 TI - Antipituitary antibodies in adults with apparently idiopathic growth hormone deficiency and in adults with autoimmune endocrine diseases. AB - The role of antipituitary antibodies (APA) in autoimmune pituitary diseases still needs to be clarified. The aim of this study was 2-fold: first, to investigate the presence of APA in adults with idiopathic or acquired GH deficiency (GHD) and in adults with autoimmune endocrine diseases; and second, to evaluate whether in autoimmune endocrine patients APA titer is correlated to the pituitary function and particularly to GH secretion. We studied 12 adults with isolated and apparently idiopathic GHD who were treated with recombinant GH in childhood (group 1a), 14 patients with adult GHD secondary to surgery for pituitary and parasellar tumors (group 1b), and 180 patients with organ-specific autoimmune diseases (group 2). APA were evaluated by indirect immunofluorescence. In all APA positive patients and in 20 APA-negative patients of group 2, GH secretion was investigated by testing its response to insulin-induced hypoglycemia (insulin tolerance test) and, when impaired, also to arginine. APA were found (at high titers) in 4 of 12 patients of group 1a (33.3%) but were absent in all patients in group 1b. APA were also found in 40 of 180 patients of group 2 (22.2%), 35 of them at low titers (group 2a) and 5 at high titers (group 2b). Twenty of the 140 autoimmune endocrine APA-negative patients studied (group 2c) and all APA positive patients at low titers (group 2a) had normal pituitary function. Conversely, all APA-positive patients at high titers (groups 1a and 2b) had a severe isolated GHD. An inverse correlation between APA titers and GH peak serum response to insulin tolerance test in autoimmune endocrine patients was observed. Our results suggest that APA, when detected at high titers, may be considered a good diagnostic tool to highlight the possible occurrence of GHD in adults with autoimmune endocrine diseases. Moreover, they may indicate an autoimmune pituitary involvement in adults with apparently idiopathic GHD, suggesting that the prevalence of autoimmune GHD is much higher than that so far considered. PMID- 12574196 TI - Ovarian steroids differentially modulate the gene expression of gonadotropin releasing hormone neuronal subtypes in the ovariectomized cynomolgus monkey. AB - In the present study, we compared the morphology and distribution of neurons expressing GnRH gene transcripts in the hypothalamus and forebrain of the cynomolgus monkey to that of the human. As in the human, three subtypes of GnRH neurons were identified. Type I GnRH neurons were small, oval cells with high levels of gene expression and were located within the basal hypothalamus. Type II GnRH neurons were small and sparsely labeled and were widely scattered in the hypothalamus, midline nuclei of the thalamus, and extended amygdala. Type III neurons displayed magnocellular morphology and intermediate labeling intensity and were located in the nucleus basalis of Meynert, caudate, and amygdala. In a second experiment, we determined the effect of estrogen or estrogen plus progesterone on the gene expression of GnRH neurons in the brains of young, ovariectomized cynomolgus monkeys. We report that hormone treatment resulted in a significant decrease in GnRH mRNA in type I neurons within the basal hypothalamus of ovariectomized monkeys. In contrast, there was no effect of hormone treatment on the gene expression of type III GnRH neurons in the nucleus basalis of Meynert. The present findings provide evidence that the increase in gene expression of type I GnRH neurons in postmenopausal women is secondary to the ovarian failure of menopause. The differential responses of type I and III GnRH neurons to hormone treatment provide additional evidence that distinct subpopulations of neurons expressing GnRH mRNA exist in the primate hypothalamus. PMID- 12574197 TI - Differential regulation of gonadotropin-releasing hormone (GnRH)I and GnRHII messenger ribonucleic acid by gonadal steroids in human granulosa luteal cells. AB - In humans, reproduction was generally believed to be controlled by only one form of GnRH (called mammalian GnRH or GnRHI). However, recently, a second form of GnRH, analogous to chicken GnRHII, was discovered in several tissues, including the human ovary. The regulation and function of GnRHI in the hypothalamus has been well studied. However, the function and regulation of GnRHI, and particularly GnRHII in the ovary, is less well understood. Because gonadal sex steroids are one of the main regulators of reproduction, we investigated, in the present study, the regulation of GnRHI and GnRHII mRNA expression by 17beta estradiol (E2) and RU486 (a progesterone antagonist) in human granulosa luteal cells (hGLCs). The levels of the mRNA transcripts encoding the two GnRH forms were examined using semiquantitative RT-PCR followed by Southern blot analysis. With time in culture, GnRHI and GnRHII mRNA levels significantly increased, by 120% and 210%, at d 8 and d 1, respectively. The levels remained elevated until the termination of these experiments at d 10. A 24-h treatment of hGLCs with E2 (10(-9) to 10(-7) M) resulted in a dose-dependent decrease and increase in mRNA expression of GnRHI and GnRHII, respectively. E2 (10(-9) M) significantly decreased GnRHI mRNA levels (by 55%) and increased GnRHII mRNA levels (by 294%). Time-course studies demonstrated that E2 (10(-9) M) significantly decreased GnRHI mRNA levels in a time-dependent manner, with maximal inhibition of 77% at 48 h. In contrast, GnRHII mRNA levels significantly increased in a time-dependent fashion, reaching a maximum level of 280% at 24 h. Cotreatment of hGLCs with E2 and tamoxifen (an E2 antagonist) reversed the inhibitory and stimulatory effects of E2 on the mRNA expression of GnRHI and GnRHII, respectively. Time- and dose dependent treatment with RU486 did not affect GnRHI mRNA levels in hGLCs. In contrast, RU486 treatment significantly increased GnRHII mRNA levels in hGLCs in a time- and dose-dependent fashion, with a maximum increase being observed at 24 h (with 10(-5)M RU486). In summary, the present study demonstrated that the expression of GnRHI and GnRHII at the transcriptional level is differently regulated by E2 and P4 in hGLCs. PMID- 12574198 TI - Homocysteine levels and C677T polymorphism of methylenetetrahydrofolate reductase in women with polycystic ovary syndrome. AB - The aim of this study was to investigate the homocysteine (Hcy) levels and the C677T polymorphism of 5,10-methylenetetrahydrofolate reductase (MTHFR), a crucial factor of the Hcy metabolism in young women with polycystic ovary syndrome (PCOS). Seventy young women with PCOS and another 70 healthy women with low folate intake were enrolled. Cases and controls were matched for age, body mass index, and allele frequency. Hcy, vitamin B(12), and folate levels were measured, and a genetic analysis of 5,10-MTHFR at nucleotide 677 was performed in all subjects. No difference in mean Hcy levels was observed between PCOS women in comparison to the control group. Considering the different MTHFR polymorphism, no significant difference was found in serum Hcy levels between subjects with PCOS and controls showing CC (10.4 +/- 3.1 vs. 9.7 +/- 2.9 micromol/liter +/- SD) and CT genotypes (10.9 +/- 3.8 vs. 11.0 +/- 3.2 micromol/liter +/- SD). In subjects with a TT homozygous state, a significant (P < 0.05) difference was observed between PCOS and control women (11.5 +/- 3.9 vs. 22.0 +/- 7.8 micromol/liter +/- SD). In conclusion, our data show that in PCOS women, the serum Hcy levels are normal, and the C677T polymorphism of MTHFR does not influence the Hcy levels like in controls. PMID- 12574199 TI - Dose-dependent effects of gonadotropin releasing hormone on matrix metalloproteinase (MMP)-2, and MMP-9 and tissue specific inhibitor of metalloproteinases-1 messenger ribonucleic acid levels in human decidual Stromal cells in vitro. AB - Matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue-specific inhibitor of matrix metalloproteinases (TIMPs), play key roles in the cyclic remodeling events that occur in the human endometrium in preparation for pregnancy. To date, the factors capable of regulating the expression of MMPs and TIMPs in the human decidua remain poorly characterized. The spatiotemporal expression of GnRH in the human endometrium during the menstrual cycle and early pregnancy suggests that this hormone may have a regulatory role in the development of this dynamic tissue. In view of these observations, we have examined the ability of GnRH to regulate MMP-2, MMP-9, and TIMP-1 mRNA levels in primary cultures of human decidual stromal cells using a quantitative competitive PCR strategy. GnRH was capable of increasing MMP-2 and MMP-9 mRNA levels in these primary cell cultures in a dose-dependent manner. The GnRH antagonist, antide, was capable of inhibiting the GnRH-mediated increase in the levels of the MMP-2 and MMP-9 mRNA transcripts present in these decidual stromal cells in a dose dependent manner. In contrast, GnRH or antide did not have a significant effect on TIMP-1 mRNA level in these primary cell cultures at any of the concentrations used in these studies. Taken together, these observations suggest that GnRH plays an integral role in human implantation, by virtue of its ability to regulate the balance between MMP and TIMP expression in decidual cells. PMID- 12574200 TI - Prolactin expression and secretion by human breast glandular and adipose tissue explants. AB - Prolactin (PRL) is a 23-kDa hormone produced by the pituitary and extrapituitary sites. The main target of PRL is the breast, where it affects cellular growth, differentiation, and milk production. Recent evidence suggests that locally produced PRL plays a role in breast tumorigenesis. Our objective was to examine PRL synthesis/release in different tissues of the human breast and determine the effect of ovarian steroids. Breast tissue, obtained from women undergoing mastectomy or breast reduction, was separated into glandular (nonmalignant) and adipose explants and incubated for 10 d. Conditioned media were analyzed for PRL by a bioassay. PRL release from glandular explants decreased by 60% from d 1-3, followed by a 4-fold increase on d 10. PRL release from adipose explants was unchanged from d 1-3 and increased more than 10-fold by d 10. PRL gene expression, determined by RT-PCR, was low on d 0 and markedly increased on d 10 in both types of explants. De novo synthesis of PRL was confirmed by metabolic labeling. Progesterone suppressed PRL release from glandular explants without affecting adipose explants. Estradiol did not alter PRL release from either tissue. In conclusion, the human breast produces and releases bioactive PRL, with a higher release rate by adipose than glandular tissue. The time-dependent rise in PRL release suggests removal from inhibitory control. Progesterone may be one of the factors that suppresses PRL production in the glandular compartment, whereas the factor(s) that regulate adipose PRL are unknown. These data suggest an autocrine/paracrine role for PRL in human glandular and adipose breast tissue. PMID- 12574201 TI - Calcium sensing receptor gene A986S polymorphism and responsiveness to calcium supplementation in postmenopausal women. AB - Recently several studies in adolescent girls or premenopausal women have implicated the calcium sensing receptor (CASR) gene A986S polymorphism in calcium and bone metabolism. However, the role of this genetic variant in postmenopausal women, specifically the development of osteoporosis, is unknown. This study reports the findings of a randomized, double-blind, placebo-controlled study of healthy postmenopausal women followed for 2 yr while taking placebo or supplementary calcium. Specifically, we examined the relationship between the CASR A986S polymorphism, bone biochemical profile, and bone mineral density at baseline and after 2 yr of treatment. We found no effect of this genetic variant in postmenopausal women at baseline or in response to calcium supplementation. These results are in contrast to those in young or premenopausal women, and they provide no support for an important role for the CASR A986S polymorphism in osteoporosis. PMID- 12574202 TI - Stimulatory effects of ghrelin on circulating somatostatin and pancreatic polypeptide levels. AB - Ghrelin, the recently identified endogenous ligand of the GH secretagogue receptor, is a gut-brain peptide with endocrine, orexigenic, and gastrointestinal effects. In rodents it increases circulating gastrin and insulin levels, whereas in man it appears to decrease insulin secretion despite a rise in blood glucose levels. The aim of the present study was to evaluate the effects of ghrelin administration on total circulating somatostatin (SS), pancreatic polypeptide (PP), and gastrin levels compared with those elicited on insulin, glucose, and GH. Eight healthy volunteers of normal weight (four women and four men) were injected with 3.3 microg/kg ghrelin or saline after an overnight fast on 2 different days. Blood was taken every 15 min for 1 h and then every 30 min for 2 h. As expected, ghrelin injection elicited a prompt GH and glucose increase with a peak at 30 min and an insulin decrease with a nadir at 60 min. Gastrin concentrations were not modified, whereas significant rises were observed in both SS (in a biphasic pattern with peaks at 15 and 120 min) and PP (which increased promptly with a peak at 15 min). A significant negative correlation was found between SS (first peak) and insulin changes (r = -0.86; P < 0.01). In conclusion, this study clearly demonstrates that ghrelin stimulates SS and PP release in man. Although the underlying mechanisms and biological significance of these pharmacological effects remain to be elucidated, a causal relationship between the SS increase and the insulin changes may be hypothesized. Finally, these findings strongly support ghrelin's postulated role in linking the endocrine control of energy balance and growth with the regulation of gastrointestinal functions. PMID- 12574203 TI - Sex hormone metabolism in prostate cancer cells during transition to an androgen independent state. AB - The progression of prostate cancer during androgen deprivation therapy is a serious clinical problem. Little is known, however, about the mechanisms behind the transition of the disease to an androgen-independent stage. In the present report, we provide evidence of substantial changes in both estrogen and androgen metabolism during the transition of cultured prostate cancer LNCaP (lymph node carcinoma of the prostate) cells. The results of enzyme activity measurements performed using HPLC suggest that, related to the transition, there exists a remarkable decrease in the oxidative 17 beta-hydroxysteroid dehydrogenase (17HSD) activity, whereas the reductive 17HSD activity seems to increase. Relative quantitative RT-PCR revealed that the decrease in oxidative activity largely coincided with the remarkable decrease in the expression of the HSD17B2 gene. Furthermore, the present data suggest that the observed increasing activity of 17HSD type 7 could lead to the increased intracellular production of 17 beta estradiol during disease progression. This was supported by the cDNA microarray screening results, which showed a considerable overexpression of several estrogen up-regulated genes in the LNCaP cell line variant that represents progressive prostate cancer. Because 17HSDs critically contribute to the control of bioavailability of active sex steroid hormones locally in the prostate, the observed variation in intraprostatic 17HSD activity might be predicted to be crucially involved in the regulation of growth and function of the organ. PMID- 12574204 TI - RU486-induced growth inhibition of human endometrial cells involves the nuclear factor-kappa B signaling pathway. AB - Previous studies indicated that the antiprogestin RU486 could directly inhibit the growth of normal and malignant human endometrial cells. However, the mechanism by which this occurs is poorly understood. In this study we explore further details of endometrial cell growth regulation by RU486. Gel shift assays using the endometrial cell line EM42 demonstrated that RU486, at concentrations ranging from 20-100 micro M, significantly stimulated the cellular binding activity of the nuclear transcription factor nuclear factor-kappa B (NF-kappa B) while having little effect on activating protein-1 (AP-1) binding. This effect on NF-kappa B binding was blocked in the presence of the NF-kappa B inhibitor, pyrrolidine dithiocarbamate (PDTC). The data also showed that the activity of RU486 on NF-kappa B binding correlated with the ability of this compound to induce apoptosis of EM42 cells. To investigate a cause and effect relationship between these two phenomena, we evaluated the effects of RU486 treatment on the expression of two apoptosis-related genes, bax and bcl-2, known to be regulated through NF-kappa B binding on their promoter. RT-PCR demonstrated that RU486 significantly induced bax mRNA levels, while suppressing mRNA of bcl-2. Alteration of these genes by RU486 was inhibited in the presence of 100 microM PDTC. Correspondingly, PDTC antagonized the ability of RU486 to inhibit the growth and induce apoptosis of EM42 cells. This study demonstrates that the inhibition of growth and apoptosis of human endometrial cells by RU486 involves stimulation of NF-kappa B binding with subsequent modulation of apoptosis regulatory genes bax and bcl-2. PMID- 12574205 TI - Differential expression of beta 1c integrin messenger ribonucleic acid and protein levels in human endometrium and decidua during the menstrual cycle and pregnancy. AB - beta(1C) and beta(1A) integrins are alternatively spliced variants of the human beta(1)-subunit; the former has been shown to inhibit cell proliferation, and the latter to promote it. Although some components of the beta(1) integrin subfamily are expressed in human endometrial and decidual cells during the menstrual cycle and early pregnancy, to date no information is available about the expression of beta(1C) integrin in endometrial and decidual tissues and its possible roles during implantation and pregnancy. To gain further insight on this subject, we have explored beta(1C) integrin expression in endometrial (proliferative, secretory, and atrophic) and decidual (from the first and third trimesters of pregnancy) tissue samples at both gene and protein levels by Northern and Western blotting analyses and by immunohistochemistry. beta(1A) protein levels were also measured in the same tissues as a control. The results of this study demonstrate that both beta(1C)- and beta(1A)-subunits are expressed in the endometrium and decidua. In the former, maximal beta(1C) expression was detected in atrophic endometria, whereas beta(1A) expression levels were increased in secretive and decreased in atrophic endometrial tissues compared with proliferative endometria. In addition, whereas beta(1A) levels were significantly increased in decidual tissues, compared with proliferative endometria, beta(1C) expression was dramatically reduced in the same tissues, thus pointing to selective down regulation of beta(1C) expression in the decidua. These data suggest that the expression of beta(1C) integrin, a very efficient inhibitor of cell proliferation, may be modulated by the maternal microenvironment and may play a fundamental role in mediating trophoblast outgrowth and migration during pregnancy. PMID- 12574206 TI - Tumor necrosis factor-alpha-induced interleukin-8 (IL-8) expression in endometriotic stromal cells, probably through nuclear factor-kappa B activation: gonadotropin-releasing hormone agonist treatment reduced IL-8 expression. AB - Endometriosis, a common disease among women of reproductive age, is characterized by the presence of endometrial-like tissue outside the uterus. We previously reported that TNFalpha promoted proliferation of endometriotic stromal cells by inducing IL-8 gene and protein expression. We hypothesize that TNFalpha may induce IL-8 production in endometriotic cells through nuclear factor-kappa B (NF kappa B) activation. Western blot analyses and electrophoretic mobility shift assays revealed that incubation with TNF alpha induced the expression of phosphorylated inhibitor kappa B (p-I kappa B) and activation of NF-kappa B in endometriotic stromal cells. The NF-kappa B inhibitor, N-tosyl-L-phenylalanine chloromethyl ketone, reduced TNFalpha-induced IL-8 gene and protein expression. The medical treatment of endometriosis with GnRH agonist (GnRHa) has been shown to induce hypoestrogenemia and reduce the observable number of endometriotic implants. We compare the expression of IL-8 gene and protein in endometriotic stromal cells of patients treated with GnRHa and those of patients without treatment before laparoscopic cystectomy for endometrioma. The addition of TNFalpha (0.1 ng/ml) significantly increased protein and gene expression of IL-8 in the cells of patients without GnRHa treatment, but this expression was not observed in the cells of patients with GnRHa. The addition of estradiol (E2; 10( 7) M) enhanced the expression of IL-8. However, in the cells of patients who received GnRHa treatment, TNFalpha and E2 did not show any significant effect. In endometriotic stromal cells without GnRHa treatment, TNFalpha and E2 increased the expression of p-I kappa B. In contrast, TNFalpha and E2 had no significant effect on the expression of p-I kappa B in cells that received GnRHa treatment. These findings demonstrate that NF-kappa B activation is critical for TNFalpha induced IL-8 expression in endometriotic stromal cells. The current study showed for the first time that GnRHa treatment attenuated the expression of IL-8 by reducing TNFalpha-induced NF-kappa B activation. PMID- 12574207 TI - Increased plasma thrombin-activatable fibrinolysis inhibitor levels in normotensive type 2 diabetic patients with microalbuminuria. AB - Hypofibrinolysis is a common finding in patients with diabetes mellitus and a risk factor for diabetic nephropathy. Recently, a new potent inhibitor of fibrinolysis, the thrombin-activatable fibrinolysis inhibitor (TAFI), has been isolated from human plasma. The possibility that TAFI also participates in the mechanism of hypofibrinolysis has not been appraised in diabetic patients with microalbuminuria. In the present study, we investigated the plasma levels of TAFI and its relation to urinary albumin excretion in normotensive diabetic patients with normo- and microalbuminuria. Thirty-nine normotensive nonobese type 2 diabetic patients (27 with normoalbuminuria, 12 with microalbuminuria) and 20 age matched normal subjects were enrolled in this study. The plasma level of thrombin antithrombin complex was significantly increased (22.1 +/- 2.6 vs. 8.3 +/- 1.0 nmol/liter; P < 0.05), whereas the D-dimer/thrombin-antithrombin complex ratio was significantly decreased (15.7 +/- 1.4 vs. 26.5 +/- 2.2; P < 0.05), showing the occurrence of hypercoagulability and hypofibrinolysis in diabetic patients. The plasma level of TAFI in diabetic patients was significantly elevated, compared with normal subjects (147.4 +/- 11.6 vs. 99.5 +/- 4.9%; P < 0.05). The plasma level of TAFI in diabetic patients with microalbuminuria was significantly higher than the level in diabetic patients with normoalbuminuria (194.1 +/- 24.5 vs. 128.8 +/- 12.3%; P < 0.02) or normal subjects (194.1 +/- 24.5 vs. 99.5 +/- 4.9%; P < 0.005). Univariate analysis showed that the plasma TAFI levels are significantly and proportionally correlated with urinary albumin excretion rate (r = 0.58; P < 0.005) and with plasma soluble thrombomodulin level, a marker of endothelial cell damage, in all diabetic patients (r = 0.42; P < 0.01). These data suggest that increased plasma level of TAFI may be involved in the mechanism of vascular endothelial damage in patients with type 2 diabetes mellitus. PMID- 12574208 TI - Glucose stimulates pulsatile insulin secretion from human pancreatic islets by increasing secretory burst mass: dose-response relationships. AB - Insulin is secreted almost exclusively in discrete bursts, and physiological regulation is accomplished by modulation of the pulse mass. How the integrity of contiguous anatomic structures in the human pancreas (islets, splanchnic innervation, exocrine tissue, local hormones) directs the coordinated insulin secretion is not known. We posed the hypothesis that glucose stimulates insulin secretion from isolated human islets by an amplification of insulin pulse mass with no change in pulse frequency and that the glucose dose-response curve for the regulation of insulin pulse mass mirrors that recognized in vivo. Islets from five nondiabetic cadaveric donors were perifused in a recently validated perifusion system at 4 mM and subsequently at 8, 12, 16, or 24 mM glucose. The effluent was collected in 1-min intervals and used for the measurement of insulin (ELISA). Pulsatile insulin secretion was analyzed by deconvolution analysis. Total insulin secretion increased progressively (P < 0.0001). This augmentation was due to amplified pulse mass (3-fold, 24 mM vs. 4 mM glucose; P < 0.0001) with no change in pulse interval (approximately 4 min). Pulsatile insulin secretion was stimulated most effectively in a physiologic concentration range of 4-8 mM. The islet insulin content was significantly correlated to the magnitude of first and second phase insulin secretion (P < 0.0001). The quantifiable orderliness of pulsatile insulin secretion rose with escalating glucose concentration (P = 0.02). In conclusion, glucose stimulates pulsatile insulin secretion from isolated human islets by amplification of insulin pulse mass without altering pulse interval. The in vitro concentration-response relationship is comparable with that observed in vivo. These data imply that transplanted human islets should be able to reproduce glucose-regulated insulin secretion as observed in the intact human pancreas. PMID- 12574209 TI - Analysis of cancer/testis antigens in sporadic medullary thyroid carcinoma: expression and humoral response to NY-ESO-1. AB - Cancer/testis antigens (CTA) are tumor-associated antigens expressed during ontogenesis, in a number of solid tumors but not in normal tissues except testis. Most of these CTA are highly immunogenic, eliciting a humoral and cellular response in the patients with advanced cancer, and are useful for tumor-specific immunotherapy. Medullary thyroid carcinoma (MTC) is a neoplasm derived from the parafollicular cells of the thyroid and occurs in either a sporadic or a familial form. In the present study, we examined by RT-PCR the expression of a number of genes encoding CTA in 23 surgical samples of sporadic MTC. Among the 11 cDNA antigens examined, RAGE, MAGE-4, and GAGE 1-2, were not expressed in any of the tissues. SSX 2 was present only in one tissue, whereas BAGE, GAGE 1-6, MAGE-1, MAGE-2, MAGE-3, and SSX 1-5 were detected in two to five samples. NY-ESO-1 cDNA was the most frequent, being detected in 15 of 23 examined samples (65.2%). Six (26.1%) tissues did not express any CTA-specific mRNA, whereas 10 tumors expressed only one gene (43.5%), 3 (21.4%) expressed 2 genes, and 4 displayed a broad CTA gene expression. NY-ESO-1 expression in primary MTC tissues significantly correlated with tumor recurrence. The presence of specific anti-NY ESO-1 antibodies was searched in the sera of MTC-affected patients examined by ELISA using recombinant NY-ESO-1 protein. A humoral response against this CTA was detected in 6 of 11 NY-ESO-1 expressing patients (54.5%), and in 1 of 6 patients with NY-ESO-1-negative tumor. No anti-NY-ESO-1 antibodies were detected in healthy subjects (n = 17). The presence of anti-NY-ESO-1 antibodies was searched also in the sera of MTC affected patients whose tissues were not available for CTA analysis. Anti-NY-ESO-1 antibodies were present in 15 of 42 sera (35.7%), demonstrating that MTC is a neoplasm frequently associated with humoral immune response to NY-ESO-1. Serological survey may be useful as a way to identify patients with humoral immune response to NY-ESO-1 that provide a new attractive target for vaccine-based immunotherapy of MTC. PMID- 12574211 TI - p21 Waf-1 (Cip-1) enhances apoptosis induced by manumycin and paclitaxel in anaplastic thyroid cancer cells. AB - Our previous studies demonstrated that manumycin (a farnesyltransferase inhibitor) enhanced the antineoplastic activity and induction of apoptosis when combined with paclitaxel against anaplastic thyroid cancer cells. We found that manumycin induces endogenous expression of p21 Waf-1 in anaplastic thyroid cancer cells. Manumycin increased the activity of the p21promoter, the level of p21mRNA, and the amount of p21 protein. We hypothesized that p21 had a proapoptotic effect in cells treated with manumycin, or paclitaxel, or both agents. By measuring viability and caspase-3 activity, we found that stably transfected KAT-4 cells with p21 cDNA under the control of a metallothionein promoter were more sensitive to manumycin alone, paclitaxel alone, and manumycin plus paclitaxel when p21was induced. The increased sensitivity of the cells with induced p21 was associated with an increase in caspase-3 activity (i.e. apoptosis). We also found that cells with both p21 alleles deleted were less sensitive to manumycin plus paclitaxel than its wild-type parent cells. Expression of p21 per se did not induce apoptosis but enhanced the cytotoxic effects of manumycin and paclitaxel. These findings suggested that p21 might be required to maintain cell sensitivity to the cytotoxic effects of manumycin and paclitaxel. PMID- 12574210 TI - Growth differentiation factor-9 induces Smad2 activation and inhibin B production in cultured human granulosa-luteal cells. AB - The TGF beta family member growth differentiation factor-9 (GDF-9) is an oocyte derived factor that is essential for mammalian ovarian folliculogenesis. GDF-9 mRNAs have been shown to be expressed in the human ovarian follicle from the primary follicle stage onward, and recombinant GDF-9 has been shown to promote human ovarian follicle growth in vitro. In this study with primary cultures of human granulosa-luteal (hGL) cells, we investigated whether recombinant GDF-9 activates components of the Smad signaling pathways known to be differentially activated by TGF beta and the bone morphogenetic proteins (BMPs). As with TGF beta, GDF-9 treatment caused the phosphorylation of endogenous 53-kDa proteins detected in Western blots with antiphospho-Smad2 antibodies (alpha PS2). However, unlike BMP-2, GDF-9 did not activate the phosphorylation of antiphospho-Smad1 antibody (alphaPS1)-immunoreactive proteins in hGL cells. Infection of hGL cells with an adenovirus expressing Smad2 (Ad-Smad2) confirmed that GDF-9 activates specifically phosphorylation of the Smad2 protein. Infection of hGL cells with Ad Smad7, which expresses the inhibitory Smad7 protein, suppressed the levels of both GDF-9-induced endogenous and adenoviral alpha PS2-reactive proteins. Furthermore, GDF-9 increased the steady state levels of inhibin beta(B)-subunit mRNAs in hGL cells and strongly stimulated the secretion of dimeric inhibin B. Again, Ad-Smad7 blocked GDF-9-stimulated inhibin B production in a concentration dependent manner. We identify here for the first time distinct molecular components of the GDF-9 signaling pathway in the human ovary. Our data suggest that GDF-9 mediates its effect through the pathway commonly activated by TGF beta and activin, but not that activated by many BMPs. The results are also consistent with the suggestion that in addition to endocrine control of inhibin production by gonadotropins, a local paracrine control of inhibin production is likely to occur via oocyte-derived factors in the human ovary. PMID- 12574212 TI - Activation of mitogen-activated protein kinase pathway by keratinocyte growth factor or fibroblast growth factor-10 promotes cell proliferation in human endometrial carcinoma cells. AB - Fibroblast growth factors (FGFs) exert diverse effects resulting from their interaction with cognate receptors on target cells. Our current study was designed to examine the local production and action of two specific stromal epithelial cell mediatory factors, keratinocyte growth factor (KGF) and FGF-10, in human endometrial carcinoma cells. The RT-PCR method was used to determine gene expression of KGF, FGF-10, and KGF receptor in human endometrial carcinoma cells (HEC-1) and human endometrial stromal cells. KGF mRNAs were expressed in both of these cell types. On the other hand, FGF-10 mRNA was detected only in the endometrial stromal cells, and KGF receptor mRNA was observed in the HEC-1 cells. The novel finding of the present study is that KGF is expressed in carcinoma cells and FGF-10 is expressed in human endometrial stromal cells. The distinct phosphorylation of ERK-1 and -2 (ERK1/2), which are members of the MAPK family, was observed when HEC-1 cells were treated with KGF or FGF-10. KGF and FGF-10 could induce the prompt phosphorylation of ERK1/2 and consequently stimulate DNA synthesis. KGF and FGF-10 did not activate the phosphorylation of Akt, protein kinase C, or signal transducer and activator of transcription-3. Blocking the MAPK pathway with the specific methyl ethyl ketone 1/2 inhibitor (U0126) completely neutralized the enhancement of cell proliferation induced by KGF and FGF-10. In addition, KGF and FGF-10 activated expressions of downstream nuclear transcription factors, such as Elk-1 and c-myc, but not c-fos. These results demonstrate for the first time that KGF and FGF-10 are capable of stimulating the growth of endometrial carcinoma cells via activating MAPK pathway through autocrine/paracrine fashion. PMID- 12574213 TI - Atypical Bartter syndrome with sensorineural deafness with G47R mutation of the beta-subunit for ClC-Ka and ClC-Kb chloride channels, barttin. AB - Bartter syndrome comprises several related renal tubular disorders including classic Bartter, infantile Bartter (IBS), and Gitelman syndrome. A new distinct group in Bartter syndrome accompanied by sensorineural deafness (BSND) has been identified among the IBS patients. Recently a gene encoding an essential beta subunit for ClC chloride channels, named barttin, with several mutations of the gene as the cause of BSND, has been described. We have observed a male who had not been diagnosed as Bartter syndrome until 28 yr because of a mild clinical manifestation. The patient was affected with congenital deafness, which urged us to analyze his gene for barttin, and a mutation G47R, which was previously reported, has been identified. However, the clinical feature in the patient lacking the characteristic symptoms of IBS such as polyhydramnios, premature labor, or severe salt loss in neonatal period contrasts with that of the typical BSND patients described so far in the literature. This might be due to a less severe loss of function of barttin induced by G47R mutation, compared with others, and our observation seems to suggest a possibility of the prevalence of mild form BSND with various levels of barttin dysfunction among patients with congenital deafness of unknown origin. PMID- 12574214 TI - The role of Mullerian inhibiting substance in the evaluation of phenotypic female patients with mild degrees of virilization. AB - Mullerian inhibiting substance (MIS) is a sexually dimorphic gonadal hormone with proven efficacy in the evaluation of boys with cryptorchidism and children with intersex conditions. We examined the role of MIS determination in the evaluation of 65 phenotypic females with mild virilization. Among the 28 subjects with MIS values elevated above the normal female range, all had abnormal gonadal tissue: ovotestes in 11, testes in 7, dysgenetic gonads in 7, and MIS-secreting ovarian tumors in 3. Among the 37 children with serum MIS in the normal female range, 19 had detectable MIS and 18 had unmeasurable MIS. In the former group with measurable but normal female MIS values, 16 subjects had ovaries, 1 had an ovotestis, and 1 had dysgenetic gonads containing testicular elements. Of 18 children with undetectable MIS values, 16 had ovaries and 2 had ovarian dysgenesis. In this study, elevation of serum MIS above the normal female range was consistently associated with the presence of testicular tissue or MIS- secreting tumors, mandating additional evaluation and surgical exploration. A value within the normal female range in a virilized patient did not exclude dysgenetic testicular tissue or ovotestis, whereas undetectable values were consistent with the absence of testicular tissue. PMID- 12574215 TI - Multiple indexes of lipid availability are independently related to whole body insulin action in healthy humans. AB - An increase in muscle lipid content has been postulated to relate closely to the evolution of insulin resistance. We aimed to test whether the multiple indexes of lipid supply within man [namely, circulating triglycerides, skeletal muscle triglycerides (SMT), total and central fat mass, and circulating leptin] were independent predictors of insulin resistance, or whether triglycerides from different sources are additive in their influence on whole body insulin sensitivity. Whole body insulin sensitivity, body composition, and SMT content were determined in 49 sedentary, nondiabetic males (age, 20-74 yr; body mass index, 20-38 kg/m(2)). Insulin sensitivity was inversely associated with central abdominal fat (r(2) = 0.38; P < 0.0001), total body fat (r(2) = 0.21; P = 0.0003), SMT content (r(2) = 0.16; P = 0.005), and fasting triglycerides (r(2) = 0.24; P = 0.0003), nonesterified free fatty acid (r(2) = 0.19; P = 0.002), and leptin (r(2) = 0.35; P < 0.0001) levels. However, only central abdominal fat was significantly related to SMT content (r(2) = 0.10; P = 0.03). SMT content, circulating triglycerides, and measurements of total or central adiposity were independent predictors of whole body insulin sensitivity. PMID- 12574216 TI - Estrogen-metabolizing gene polymorphisms, but not estrogen receptor-alpha gene polymorphisms, are associated with the onset of menarche in healthy postmenopausal Japanese women. AB - Both onset and cessation of menstruation have strong genetic inclination. We aimed to identify genetic factors influencing the onset of menarche and natural menopause in a Japanese population by investigating the polymorphisms of estrogen receptor-alpha and estrogen-metabolizing enzyme genes. Three hundred seventeen postmenopausal Japanese women, aged 46 yr and over, were enrolled in this study under informed consent. Genomic DNA was extracted from peripheral leukocytes, and PCR-based restriction fragment length polymorphism assays were used to determine estrogen receptor-alpha: PvuII, XbaI, and estrogen-metabolizing enzymes; CYP17, estrogen biosynthesis (high activity, A2/A2, CYP1A1), hydroxylation (high inducibility, vt/vt, and COMT), inactivation (low activity, L/L) genotypes. There were no significant differences in ages at menarche and natural menopause or years of menstruation among each PvuII or XbaI genotype and seven combinations of PvuII and XbaI genotypes. We found that ages at menarche in women with A1/A2 (higher activity of CYP17; 13.6 +/- 1.2 yr) were significantly earlier than in those with A1/A1 (lower activity of CYP17; 14.1 +/- 1.3 yr). There were no significant differences in age at natural menopause and years of menstruation among each CYP17, CYP1A1, or COMT genotype. The small sample size of each combination of estrogen-metabolizing genotypes made it impractical to evaluate the effects of the interdependency of each genotype, including extreme genotype categories such as A2/A2L/Lvt/vt vs. A1/A1H/Hwt/wt genotypes, on ages at menarche and/or natural menopause. The results suggest that the estrogen-metabolizing CYP17 genotype influences age at menarche in healthy postmenopausal Japanese women. PMID- 12574217 TI - A new nonisotopic, highly sensitive assay for the measurement of human placental growth hormone: development and clinical implications. AB - Human placental GH (hGH-V) is a variant of pituitary hGH (hGH-N) synthesized and secreted by syncytiotrophoblasts during pregnancy. It differs from hGH-V by only 13 amino acid residues, which makes difficult a specific measurement of hGH-V without interference from hGH-N. To overcome the analytical difficulties, we produced new high affinity monoclonal antibodies specific for hGH-V. Precise screening and epitope mapping allowed identification of a pair of monoclonal antibodies suitable to establish a highly sensitive assay for hGH-V measurement. In a prospective, longitudinal study involving 84 normal pregnancies, we measured maternal concentrations of hGH-V, leptin, IGF-I, and cord blood IGF-I. hGH-V was detectable as early as gestational week (GW) 7. Mean concentrations of hGH-V increased from 0.9 +/- 0.5 microg/liter (GW 7-13) to 2.8 +/- 0.9 microg/liter (GW 18-22), 7.3 +/- 2.6 microg/liter (GW 28-32), and 13.0 +/- 9.6 (GW 37-41). A negative correlation was found between prepregnancy body mass index and hGH-V concentrations from GW 28 onward. Peak hGH-V levels occurred at wk 36.5 +/- 2.6 and were significantly lower in obese (P = 0.029) and higher in underweight (P = 0.035) mothers compared with those in mothers of normal weight. The increase in hGH-V between GW 18-22 and GW 28-32 was negatively correlated to the increase in maternal leptin during this period (P = 0.027). Maternal IGF-I concentrations were correlated to those of hGH-V from GW 18 onward (P = 0.039). The strongest correlation was found at GW 28-32 (P = 0.001). Furthermore, maternal hGH-V concentrations in late pregnancy correlated with cord blood IGF-I (P = 0.025) and size of the newborn (P = 0.017). These results, obtained by a new, highly sensitive hGH-V-specific immunoassay, highlight the importance of maternal hGH-V in the regulation of maternal and fetal IGF-I. In addition, the results indicate that maternal weight has a major impact on circulating concentrations of hGH-V. PMID- 12574218 TI - Dietary composition in restoring reproductive and metabolic physiology in overweight women with polycystic ovary syndrome. AB - Overweight women with polycystic ovary syndrome (PCOS) were randomized to a high protein (HP; 40% carbohydrate and 30% protein; n = 14) or a low protein (LP; 55% carbohydrate and 15% protein) diet (n = 14). The intervention consisted of 12 wk of energy restriction (approximately 6000 kJ/d), followed by 4 wk of weight maintenance. Pregnancies (two HP and one LP); improvements in menstrual cyclicity, lipid profile, and insulin resistance (as measured by the homeostasis model); and decreases in weight (7.5%) and abdominal fat (12.5%) occurred independently of diet composition. Improvements in menstrual cyclicity were associated with greater decreases in insulin resistance and fasting insulin (P = 0.011). On the LP diet, high density lipoprotein cholesterol decreased 10% during energy restriction (P = 0.008), and the free androgen index increased 44% in weight maintenance stages (P = 0.027). Weight loss leads to improvements in cardiovascular and reproductive parameters potentially mediated by improvements in surrogate measures of insulin resistance. An HP weight loss diet may result in minor differential endocrine and metabolic improvements. PMID- 12574219 TI - A novel IVS2 -2A>T splicing mutation in the GH-1 gene in familial isolated growth hormone deficiency type II in the spectrum of other splicing mutations in the Russian population. AB - Isolated GH deficiency (IGHD) is characterized by genetic heterogeneity, both in familial and sporadic cases. To determine if this statement can be applied to the Russian population, we performed screening for mutations in the GH-1 gene in children living in Russia with IGHD. Twenty-eight children from 26 families with total IGHD were studied. DNA fragments, covering each of four (2-5) exons of GH-1 were amplified using PCR. Single-strand conformation polymorphism analysis followed by direct DNA sequencing identified five heterozygous mutations of splicing in intron 2, intron 3, and exon 4 of GH-1; three of them were not previously reported. We concentrated here on dominant-negative mutations causing IGHD type II, which were as follows: 1) A>T transversion of the second base of the 3'-acceptor splice site of intron 2 (IVS2 -2A>T); 2) T>C transition of the second base of the 5'-donor splice site of intron 3 (IVS3 +2T>C); 3) G>A transition of the first base of the 5'-donor splice site of intron 3 (IVS3 +1G>A). Our data indicate allelic heterogeneity of IGHD type II (IGHD II). However, all mutations in Russian IGHD II patients affect splicing, a striking difference from the mutation spectrum of other IGHD forms. The IVS2 -2A>T mutation is the first identified mutation in intron 2 of GH-1. The 5'-donor splice site of intron 3 of GH-1 is a mutational hot spot, and the IVS3 +1G>A mutation can be considered to be a common molecular defect in IGHD II in Russian patients. PMID- 12574220 TI - Aging-related changes in in vivo release of growth hormone-releasing hormone and somatostatin from the stalk-median eminence in female rhesus monkeys (Macaca mulatta). AB - GH release decreases with aging in primates. However, it is unclear whether the age-related decrease in GH release is due to a decrease in stimulatory GHRH or an increase in inhibitory somatostatin (SS) from the hypothalamus. In the present study, we measured the release of GHRH and SS in the stalk-median eminence of conscious aged (n = 7, 27.0 +/- 0.7 yr old) and young adult female monkeys (n = 12, 5.0 +/- 0.3 yr old) using the push-pull perfusion method. Mean GHRH levels during morning (0600-1200 h) and evening (1800-2400 h) in aged monkeys were 3- to 4-fold lower than in young monkeys. Pulse analysis indicated that pulse frequency, pulse amplitude, and baseline GHRH release in aged monkeys were much lower than in young adults. In contrast, mean SS levels in aged monkeys during mornings and evenings were 2-fold higher than in young monkeys. Pulse analyses indicated that amplitude and baseline levels of SS were significantly higher in aged monkeys than in young adults. There were no significant changes in the pulse frequency of SS release. Therefore, the aging-related decrease in GH release is due to a substantial decrease in GHRH release and an increase in SS release from the hypothalamus. PMID- 12574221 TI - Two common naturally occurring mutations in the human gonadotropin-releasing hormone (GnRH) receptor have differential effects on gonadotropin gene expression and on GnRH-mediated signal transduction. AB - Studies of naturally occurring human GnRH receptor (GnRHR) mutants may provide a useful approach to dissecting the signal transduction pathways involved in mediating the effects of GnRH. We have analyzed two common mutations in the GnRHR, corresponding to amino acid substitutions Gln106Arg and Arg262Gln, for their effects on the stimulation of gonadotropin subunit and GnRHR gene expression by GnRH. Despite similar impairment of GnRH-stimulated inositol phosphate production, dose-response analyses indicated that Gln106Arg and Arg262Gln both reduced the sensitivity of the FSH beta gene promoter to a greater extent than LH beta or alpha GSU, suggesting the involvement of more than one signaling pathway. Furthermore, although the sensitivities of the LH beta and FSH beta gene promoters to GnRH were similarly affected by both mutants, alpha GSU sensitivity was decreased to a greater extent by Arg262Gln than by Gln106Arg. Similarly, GnRHR gene promoter sensitivity was significantly reduced only by Arg262Gln. To further characterize the differential downstream effects of these mutant GnRHRs, we investigated their effects on additional signal transduction pathways. The mutant receptors differentially affected GnRH-mediated activation of the ERK pathway and GnRH stimulation of cAMP response element-mediated transcription. These results indicate that measurement of inositol phosphate production alone may not be adequate for assessing mutant GnRHR function and additional signal transduction pathways may better reflect physiologically relevant effects. The differential stimulation of LH beta, FSH beta, and alpha GSU gene expression may contribute to the varied phenotypes observed among patients harboring these mutations. PMID- 12574222 TI - Contractile activity of human decidual stromal cells. AB - We previously demonstrated that human decidual stromal cells (DSC), the main cellular component of the decidua, are similar in antigen phenotype and structure to myofibroblasts, cells with contractile activity. In this work we isolated and maintained DSC in fibroblast medium, in which these cells show a stable phenotype similar to that of DSC in vivo. Flow cytometric observations showed that most DSC expressed alpha-smooth muscle (alpha-SM) actin, an intermediate filament that is considered a marker of myofibroblasts and is responsible for the contractile activity of these cells. alpha-SM actin mRNA was detected by RT-PCR in these cells. The contractile activity of DSC was determined by the gel contraction assay; we found that TGF beta 1 and platelet-derived-growth factor, cytokines that are known to be inducers of myofibroblast contractility, also induced contractility of DSC. IL-2, a Th1 cytokine-related with spontaneous abortion, also activated DSC contractility. Our results confirmed that DSC are phenotypically and functionally related with myofibroblast. PMID- 12574223 TI - Splanchnic release of ghrelin in humans. AB - Ghrelin is a novel polypeptide identified in rat stomach, and it increases GH release, gastric motility, and appetite and modulates many other organ functions. It has been reported that ghrelin may be released from a variety of other tissues, but the absolute contribution of the splanchnic bed remains to be defined. We quantified the splanchnic output of ghrelin in 22 healthy people after an overnight fast, with indwelling catheters in the femoral artery and hepatic vein. Splanchnic ghrelin output was calculated by multiplying veno arterial difference in ghrelin concentration with splanchnic plasma flow (measured by indicator dye dilution technique). Plasma ghrelin was measured using (125)I-labeled ghrelin and rabbit polyclonal antibody raised against octanoylated human ghrelin. Ghrelin concentrations in the artery and in the hepatic veins were 960 +/- 82 pg/ml and 1102 +/- 90 pg/ml (P < 0.001), respectively. The veno arterial concentration difference was 143 +/- 38 pg/ml, amounting to 15% of the arterial concentration. The splanchnic output of ghrelin was 141 +/- 43 ng/min (P < 0.003). Assuming that the half-life of ghrelin is less than 60 min, the splanchnic output would explain the entire amount of circulating ghrelin in the postabsorptive state. We conclude that a substantial amount of ghrelin is being released from the splanchnic bed in the postabsorptive state in healthy human subjects and that splanchnic bed is the major source of circulating ghrelin in humans. PMID- 12574224 TI - Hyperthyroidism is associated with suppressed circulating ghrelin levels. AB - Ghrelin stimulates GH secretion as well as appetite and food intake. To explore whether ghrelin is involved in the regulation of appetite and body weight in hyperthyroidism, circulating ghrelin levels were measured in nine hyperthyroid patients before and after medical treatment and compared with those in eight healthy control subjects. All participants were studied in the postabsorptive state and during a 3-h euglycemic hyperinsulinemic clamp. Before treatment the patients had 3- to 5-fold elevations of T(3), and during treatment the patients gained 5 kg of body weight. Ghrelin levels were decreased in hyperthyroidism both in the fasting state (hyperthyroid, 1080 +/- 195 pg/ml; euthyroid, 1480 +/- 215 pg/ml; P = 0.03) and during clamp (hyperthyroid, 833 +/- 150 pg/ml; euthyroid, 1210 +/- 180 pg/m; P = 0.02). After treatment, ghrelin levels did not differ from those in control subjects. In all three study groups the clamp significantly reduced ghrelin levels compared with fasting levels. In conclusion, ghrelin levels are reduced in hyperthyroidism and become normalized by medical antithyroid treatment. Hyperinsulinemia suppresses ghrelin regardless of thyroid status. Ghrelin is not a primary stimulator of appetite and food intake in hyperthyroidism, and the mechanisms underlying the suppressive effect of hyperthyroidism on ghrelin secretion remain unclear. PMID- 12574225 TI - Leukemia inhibitory factor regulates prolactin secretion in prolactinoma and lactotroph cells. AB - Leukemia inhibitory factor (LIF) stimulates the hypothalamic-pituitary-adrenal axis, and transgenic mice overexpressing pituitary LIF develop Cushing-like syndrome accompanied by reduced prolactin (PRL) expression. Effects of LIF were, therefore, tested on PRL expression in human prolactinomas and normal and tumorous rat lactotrophs. Normal human pituitary tissue expressed LIF, as well as the LIF receptor (LIFR) signaling subunits, gp130 and LIFR. Although all of 19 prolactinomas tested expressed gp130 and LIFR subunit mRNA and immunoreactive protein, only 3 of 19 prolactinomas expressed LIF mRNA. All of four prolactinomas had no detectable LIF immunoreactivity, in contrast to all other pituitary tumor types that expressed LIF. LIF (5 nM) treatment reduced PRL secretion in primary human prolactinoma cultures by up to 42% (P < 0.0005), an effect that was surprisingly blocked by sulpiride, a D2-like dopamine receptor antagonist. LIF (5 nM) also suppressed PRL levels in primary rat pituitary cultures by 70% (P < 0.005), and in rat MMQ pituitary tumor cells, and this effect was also reversed by sulpiride (200 micro M). D2R agonist suppression of PRL was not additive with LIF. GH levels in these cells were unchanged by LIF, consistent with a selective effect on PRL. Transfection of human long-form D2R into an LIF-resistant MMQ cell clone restored LIF responsiveness. These results show that even though human prolactinomas express gp130 and LIFR, and respond to exogenous LIF, albeit less than normal lactotrophs, they are largely devoid of LIF. These observations indicate a role for LIF in loss of autocrine PRL suppression contributing to unrestrained prolactinomas PRL secretion. Moreover, PRL suppression by LIF may be mediated by gp130 and D2R interaction. PMID- 12574226 TI - Role of local 11 beta-hydroxysteroid dehydrogenase type 2 expression in determining the phenotype of adrenal adenomas. AB - It is not understood why some adrenal adenomas are nonfunctional and others with similar histopathology cause preclinical or overt Cushing's syndrome. Two isozymes of 11 beta-hydroxysteroid dehydrogenase, types 1 and 2 (HSD11B1 and HSD11B2), are known to modulate glucocorticoid levels in other tissues and might influence circulating levels of active and inactive glucocorticoids if they were expressed in adrenal adenomas. We determined levels of expression of these isozymes in normal adrenals and 61 adrenal adenomas by quantitative competitive RT-PCR and immunohistochemistry. There were no differences in HSD11B1 mRNA levels among adrenal tumor groups. HSD11B2 mRNA levels were high in nonfunctioning adenomas and preclinical Cushing's adenomas compared with levels in control adrenals or in adenomas causing overt Cushing's syndrome. HSD11B2 immunoreactivity was not detected in control adrenals, but was observed in more than half of these tumors. When nonfunctioning adenomas and those causing preclinical and overt Cushing's syndrome were considered as a single group, HSD11B2 mRNA levels were strongly correlated with the ratio of plasma cortisone to cortisol, and a simple model incorporating adrenal HSD11B2 expression and tumor size as variables could predict more than 50% of the interindividual variation in plasma cortisol levels (r(2) = 0.54; P < 0.0001). Adrenal HSD11B2 may regulate levels of active and inactive glucocorticoids in the systemic circulation under these conditions, presumably by acting in an autocrine or paracrine manner. Nonfunctioning adenomas and those causing preclinical and overt Cushing's syndrome may represent a continuum with clinical manifestations depending mainly on tumor size and HSD11B2 expression levels. PMID- 12574227 TI - Cyclic changes in the expression of steroid receptor coactivators and corepressors in the normal human endometrium. AB - To examine the sex steroid-dependent growth mechanisms of the human endometrium, the expression of steroid receptor coactivators [steroid receptor coactivator-1 (SRC-1) and p300/CREB-binding protein (p300/CBP)] and corepressors (nuclear receptor corepressor and silencing mediator for retinoid and thyroid hormone receptors) was examined by immunohistochemistry, using 50 samples of normal endometria, and was compared with that of estrogen receptors (ER), progesterone receptors (PR), and proliferation marker Ki-67. In addition, actual binding of the coactivators to ER or PR was analyzed by immunoprecipitation. The expression of SRC-1 was diffusely observed in glandular and stromal cells in the proliferative phase and drastically decreased in the secretory phase. Such change in the expression pattern of SRC-1 resembled that of ER, PR, and Ki-67. On the other hand, p300/CBP expression was relatively constant throughout the menstrual cycle, with slight predominance in the proliferative phase. The expression of corepressors nuclear receptor corepressor and silencing mediator for retinoid and thyroid hormone receptors was focal in the endometrium. Immunoprecipitation, using tissue samples of both proliferative and secretory phases, revealed the complex formation between the coactivators and receptors. Binding of SRC-1 to ER was observed in the proliferative (but not in the secretory) endometrium. In contrast, binding p300/CBP to ER was noted in the endometria of both phases. Complex formation between p300/CBP and PR was noted in the secretory endometrium, whereas that between SRC-1 and PR was not apparent. Accordingly, we showed the expression pattern of steroid receptor coactivators and corepressors in the normal endometrium. Cyclic change in the expression of SRC-1 during the menstrual cycle might be important in the estrogen-action for the glandular and stromal cells. PMID- 12574228 TI - Immunolocalization of ghrelin and its functional receptor, the type 1a growth hormone secretagogue receptor, in the cyclic human ovary. AB - Ghrelin is a novel 28-amino acid peptide identified as the endogenous ligand for the GH secretagogue receptor (GHS-R). Besides its hallmark central neuroendocrine effects in the control of GH secretion and food intake, an unexpected reproductive facet of ghrelin has recently emerged because expression of this molecule and its cognate receptor has been demonstrated in rat testis. However, whether this signaling system is present in human gonads remains to be evaluated. In this study, we have assessed the presence and cellular location of ghrelin and its functional receptor, namely the type 1a GHS-R, in the cyclic human ovary by means of immunohistochemistry using specific polyclonal antibodies. Strong ghrelin immunostaining was demonstrated in ovarian hilus interstitial cells. In contrast, ghrelin signal was not detected in ovarian follicles at any developmental stage, nor was it present in newly formed corpora lutea (CL) at very early development. However, specific ghrelin immunoreactivity was clearly observed in young and mature CL, whereas expression of the peptide disappeared in regressing luteal tissue. Concerning the cognate receptor, ovarian expression of GHS-R1a protein showed a wider pattern of tissue distribution, with detectable specific signal in oocytes as well as somatic follicular cells; luteal cells from young, mature, old, and regressing CL; and interstitial hilus cells. Of particular note, follicular GHS-R1a peptide expression paralleled follicle development with stronger immunostaining in granulosa and theca layers of healthy antral follicles. In conclusion, our results are the first to demonstrate that ghrelin and its functional type 1a receptor are expressed in the cyclic human ovary with distinct patterns of cellular location. The presence of both components (ligand and receptor) of the ghrelin signaling system within the human ovary opens up the possibility of a potential regulatory role of this novel molecule in ovarian function under physiological and pathophysiological conditions. PMID- 12574229 TI - Longitudinal studies of inhibin B levels in boys and young adults with Klinefelter syndrome. AB - The aim of the study was to investigate the longitudinal changes of inhibin B in a group of patients with Klinefelter syndrome (KS; karyotype 47,XXY) progressing through puberty and to compare them to a group of age- and puberty-matched controls. Seven boys with nonmosaic KS (karyotype 47,XXY) and 11 controls were followed with longitudinal serum inhibin B measurements every 3-12 months as they approached and entered puberty. None of the boys had significant bone age delay, and all entered puberty at the normal time and progressed through it at the expected time. In addition, 15 young adults with KS, aged 16.7-29.5 yr, were studied. We found normal levels of inhibin B in prepubertal boys with KS and controls. In patients with KS as well as controls, inhibin B increased progressively before clinical pubertal onset. However, during late puberty inhibin B levels decreased gradually to the low/unmeasurable levels observed later in adult KS, while remaining unchanged in the controls. PMID- 12574230 TI - Simian virus 40-like sequences from early and late regions in human thyroid tumors of different histotypes. AB - Simian virus 40 (SV40) sequences were investigated in human thyroid tumors of different histotypes, Graves' disease thyroid specimens, normal thyroid tissues, and peripheral blood mononuclear cells (PBMC) of healthy donors. Specific SV40 large T antigen (Tag) sequences were detected, by PCR and filter hybridization, in human thyroid tumors with a frequency ranging from 66% in papillary thyroid carcinomas (PTC) to 100% in anaplastic thyroid carcinomas (ATC). SV40 was revealed in 60% and 100% of normal thyroid tissues adjacent to PTC and ATC, respectively, but in only 10% of control normal thyroid tissues (NTT) from patients affected by multinodular goiter. Thyroid tissues from patients affected by the Graves' disease were found to be SV40 positive with a frequency of 20%. In agreement with previous investigations, the presence of SV40 sequences was detected in 25% of PBMC of healthy individuals. SV40 Tag mRNA was detected by RT PCR, whereas the viral oncoprotein was revealed by immunohistochemistry with a specific monoclonal antibody. The high prevalence of SV40 footprints in human thyroid tumors indicates that the oncogenic virus may participate as a cofactor in the onset/progression of specific human thyroid cancers. Detection of SV40 sequences in NTT adjacent to thyroid cancers suggests that the viral infection may spread from transformed cells to normal cells surrounding the tumor. The presence of the SV40 footprint in PBMC implies that blood cells are vectors of the virus in other tissues of the host. PMID- 12574232 TI - The human myometrium as a target for melatonin. AB - The circadian timing of spontaneous human deliveries results in births occurring statistically more often during the nocturnal phase of the 24-h cycle. The neuroendocrine mechanisms underlying this physiological phenomenon are not understood. In an effort to test the hypothesis that melatonin may serve as an endocrine signal for coordinating myometrial events in the human, we determined the mRNA expression of both MT1 and MT2 melatonin receptor isoforms in pregnant as well as nonpregnant myometrial biopsies by means of RT-PCR and in situ hybridization histochemistry. Additionally, we could demonstrate specific, high affinity iodomelatonin binding to myometrial tissues of both pregnant and nonpregnant women. Primary cultures of myocytes responded differentially from melatonin in terms of cAMP signaling depending on the reproductive state. These results imply that melatonin may have the potential to modulate myometrial function in the human, a finding that could open up new possibilities for the development of novel therapeutic agents. PMID- 12574231 TI - Rapid insulin-like growth factor (IGF)-independent effects of IGF binding protein 3 on endothelial cell survival. AB - Angiogenic factors, such as vascular endothelial-derived growth factor (VEGF) and IGF-I, play pivotal roles in endothelial proliferation and migration. IGF binding protein-3 (IGFBP-3) is emerging as a key regulator of cell growth and apoptosis, both as an IGF antagonist and as an independent molecule. We investigated the role of IGFBP-3 in VEGFmediated survival of human macrovascular umbilical vein endothelial cells (HUVEC). Specific commercial ELISAs quantified cell proliferation and apoptosis, and Akt phosphorylation was assessed by immunoblots and confocal microscopy. IGF-I and VEGF significantly stimulated HUVEC proliferation and survival. Addition of IGFBP-3 reversed both IGF- and VEGF induced proliferation and prevented the survival induced by these factors. The antiproliferative and proapoptotic effects of exogenous IGFBP-3 upon VEGF-induced HUVEC survival were not inhibited by blockade of the type 1 IGF receptor with alpha IR-3 immunoglobulin, which fully prevented IGF actions. An IGFBP-3 mutant, which binds IGFs normally but has a substituted mid-region domain, lost the ability to inhibit VEGF actions. VEGF-induced phosphorylation of Akt, as evident by both specific immunoblots and confocal microscopy, was significantly and rapidly reduced in the presence of IGFBP-3, as well as wortmannin. PMID- 12574233 TI - Dramatic elevation of plasma metastin concentrations in human pregnancy: metastin as a novel placenta-derived hormone in humans. AB - Metastin is a novel peptide that was recently isolated from human placenta as the endogenous ligand of an orphan heptahelical receptor, hOT7T175. Metastin has been shown to suppress the motility of hOT7T175-transfected melanoma cells; however, studies of the physiological function of metastin have begun only recently. To investigate the possibility that metastin is an endocrine peptide, we determined the immunoreactive (ir-) metastin concentration in human plasma using our newly developed, sensitive, and specific two-site enzyme immunoassay. The plasma concentrations of ir-metastin in males and females were 1.30 +/- 0.14 (n = 12) and 1.31 +/- 0.37 fmol/ml (n = 10), respectively. As metastin is known to be abundant in human placenta, the ir-metastin concentration in the maternal plasma was then determined. The ir-metastin concentrations were 1230 +/- 346 fmol/ml (n = 11) in the first trimester, 4590 +/- 555 (n = 16) in the second trimester, and 9590 +/- 1640 (n = 12) in the third trimester. On d 5 after delivery, the ir metastin concentration returned to nearly the nonpregnant level (7.63 +/- 1.33 fmol/ml; n = 10), suggesting that ir-metastin increases in pregnancy and is derived mainly from the placenta. The plasma from both nonpregnant and pregnant women showed a single ir-metastin peak at the same retention time as authentic metastin on reverse phase HPLC analysis, indicating that the major portion of the circulating metastin, as determined by our two-site enzyme immunoassay, represents endogenous metastin. Histochemical studies of human placenta localized metastin mRNA and immunoreactivity to the syncytiotrophoblasts. The present study provides evidence for metastin as a novel placenta-derived hormone in humans. PMID- 12574234 TI - Hepatocyte nuclear factor-1 alpha gene mutations and diabetes in Norway. AB - Mutations in the hepatocyte nuclear factor (HNF)-1 alpha gene cause maturity onset diabetes of the young (MODY), type 3. To estimate the prevalence of MODY3 in Norwegian diabetic pedigrees, we screened a total of 130 families for HNF-1 alpha mutations; 42 families with clinical MODY, 75 with suspected MODY, and 13 pedigrees with multiplex type 1 diabetes. Twenty-two families with clinical MODY, 15 families with suspected MODY, and one family with type 1 diabetes multiplex harbored HNF-1 alpha mutations. Thus, in about half of Norwegian families with clinical MODY, mutations in the HNF-1 alpha gene could be detected. Eight of the 18 different mutations identified were novel (G47E, T196fsdelCCAA, IVS3-1G>A, S256T, A276D, S445fsdelAG, M522V, and S531T). Haplotypes were determined for recurrent mutations, indicating a founder effect in Norway for the hot-spot mutation P291fsinsC and possibly also for P112L and R131W. To examine the molecular mechanisms underlying MODY3, we investigated the functional properties of 13 HNF-1 alpha mutations. Two mutant HNF-1 alpha proteins (R171X, R263C) were unable to bind DNA and at least five mutants (R131W, R171X, P379fsdelCT, S445fsdelAG, and Q466X) showed defective nuclear translocation. Transcriptional activation was reduced for most of the MODY3-associated mutants. Accordingly, the functional studies of HNF-1 alpha mutants indicate that beta-cell dysfunction in MODY3 is caused by loss-of-function mechanisms like reduced DNA binding, impaired transcriptional activation, and defects in subcellular localization. PMID- 12574235 TI - Variation in levels of serum inhibin B, testosterone, estradiol, luteinizing hormone, follicle-stimulating hormone, and sex hormone-binding globulin in monthly samples from healthy men during a 17-month period: possible effects of seasons. AB - To obtain information on the scale of the intraindividual variation in testicular hormone, blood samples for inhibin B determination were collected monthly in 27 healthy male volunteers during a 17-month period. In addition, the traditional reproductive hormones FSH, LH, testosterone, estradiol, and SHBG were measured. The intraindividual variation in inhibin B over the study period was, on the average, 10%, corresponding to the assay variation of the inhibin B assay, indicating that most of the observed day to day variation in inhibin B levels in men could be explained by assay variation. A seasonal variation was observed in LH and testosterone levels, but not in the levels of the other hormones. The seasonal variation in testosterone levels could be explained by the variation in LH levels. The seasonal variation in LH levels seemed to be related to the mean air temperature during the month before blood sampling, but not to the length of daylight or the hours of sunshine. In conclusion, our data showed that day to day levels of inhibin B are relatively constant in men and do not seem to be influenced by seasonal factors. In contrast, we found a seasonal variation in LH and testosterone levels in men. The peak levels of both LH and testosterone were observed during June-July, with minimum levels present during winter-early spring. Air temperature, rather than light exposure, seems to be a possible climatic variable explaining the seasonal variation in LH levels. PMID- 12574236 TI - Ret/PTC chimeric transcripts in an Irish cohort of sporadic papillary thyroid carcinoma. AB - Ret rearrangements are common in papillary thyroid carcinoma (PTC), with ret/PTC 3 commonly seen in children exposed to ionizing radiation. In this context ret/PTC-3 has been correlated with solid variant morphology, poorer prognosis, and aggressive tumor behavior. We aimed to assess the prevalence of the common ret chimeric transcripts (ret/PTC-1 and ret/PTC-3) in a group of sporadic PTC and correlate them with tumor morphology. Thyroid follicular cells were laser capture microdissected from sections of archival PTC (n = 28). Total RNA was extracted and analyzed for expression of glyceraldehyde 3-phosphate dehydrogenase, ret/PTC 1, and ret/PTC-3 using TaqMan PCR. Ret/PTC rearrangements were detected in 60% of PTCs. Specifically transcripts of ret/PTC-1 and ret/PTC-3 were detected in 43% and 18% of PTCs, respectively. Ret/PTC-3 was detected in only follicular variant subtype (60%) and was not detected in classic PTC. One case of tall cell variant demonstrated chimeric expression of both ret/PTC-1 and ret/PTC-3 transcripts within the same tumor. This study demonstrated a high prevalence of the two common ret rearrangements in an Irish cohort of PTCs. A correlation of tumor morphology with these common ret rearrangements is suggested. PMID- 12574237 TI - Circulating ghrelin concentrations in the polycystic ovary syndrome. AB - Ghrelin is a novel gastric peptide which has orexigenic and adipogenic properties. Circulating ghrelin concentrations are influenced by nutritional status and, probably, regulate food intake and body weight. Obesity is a common feature in women with polycystic ovary syndrome (PCOS). To investigate the relationship of circulating ghrelin concentrations to the hormonal and metabolic features of PCOS women, plasma ghrelin and several hormone concentrations were evaluated in thirty-three women with PCOS and in thirty-two healthy women matched for age and body mass index (BMI). Plasma ghrelin concentrations were similar between the PCOS (179 +/- 27, pmol/l +/- SEM) and the control (181 +/- 24, pmol/l +/- SEM) groups. In both groups, there was a significant (P < 0.001) inverse correlation between fasting ghrelin concentrations and BMI (PCOS: r = -0.45; CONTROLS: r = -0.59). Multivariate regression analysis did not demonstrate any correlation (P = NS) between fasting ghrelin concentrations and the other hormone levels in the PCOS patients. In conclusion, our data demonstrate that in women with PCOS plasma ghrelin concentrations are not different from those of weight matched controls and are inversely correlated with BMI. There is no relationship between circulating ghrelin and the abnormal hormonal pattern of the PCOS syndrome. PMID- 12574238 TI - Flutamide counteracts the antiproliferative effects of antiprogestins in the primate endometrium. AB - In addition to blocking progesterone (P) action, antiprogestins (APs) also inhibit estrogen-stimulated endometrial cell proliferation in nonhuman primates and women. This effect is paradoxical because APs do not bind to estrogen receptors (ER), and AP + estradiol (E(2)) treatment leads to elevation of ER levels, a recognized action of estrogen in the endometrium. Recently, we showed that APs (RU 486, ZK 137 316 and ZK 230 211) also elevate endometrial androgen receptor (AR) in macaques and women and we hypothesized that over expression of AR may play a role in the antiproliferative actions of APs. We now report that cotreatment with the specific antiandrogen flutamide (FLU) blocked the suppressive effects of APs on estrogen action in the endometrium. We treated ovariectomized rhesus macaques with either E(2) alone, E(2) + ZK 137 316 or E(2) + ZK 137 316 + FLU daily for 28 days. Endometrial wet weight (mean +/- SE) from one-half of the endometrium was as follows: E(2)-treated controls, 360 +/- 32 mg; E(2) + ZK 137 316, 64 +/- 10 mg; and E(2) + ZK 137 316 + FLU, 265 +/- 92 mg (P < 0.05). Mean mitoses/1000 epithelial cells +/- SE was: E(2) alone, 6.25 +/- 0.6; E(2) + ZK 137 316, 0.3 +/- 0.25; and E(2) + ZK 137 316 + FLU, 5.1 +/- 3.8 (P < 0.05). FLU also blocked the hyalinizing degradation of the spiral arteries typically induced by APs. These results indicate that many of the antiendometrial effects of APs in primates may involve the AR. PMID- 12574240 TI - Methodological considerations regarding the use of galectin-3 expression analysis in preoperative evaluation of thyroid nodules. PMID- 12574241 TI - Clostridium difficile testing: after 20 years, still challenging. PMID- 12574242 TI - PCR assay for identification of histoplasma capsulatum based on the nucleotide sequence of the M antigen. AB - The major diagnostic antigens of Histoplasma capsulatum var. capsulatum are the H and M antigens, pluripotent glycoproteins that elicit both humoral and T-cell mediated immune responses. The gene encoding the M antigen has previously been sequenced, and its sequence has significant overall homology to those of the genes for fungal catalases. Regions of the M-antigen gene with little or no homology were used to design four oligonucleotide sequences for application in the PCR detection and identification of H. capsulatum var. capsulatum. The PCR correctly identified the 31 H. capsulatum var. capsulatum strains isolated from human, animal, and soil specimens and 1 H. capsulatum var. duboisii isolate. PCR products of 111 and 279 bp were amplified with primers Msp1F-Msp1R and Msp2F Msp2R, respectively. No amplification product was obtained from DNA extracted from an H. capsulatum var. farciminosum isolate. The specificity of the PCR with the M-antigen-derived primers was confirmed by the total absence of amplification products when genomic DNA from Paracoccidioides brasiliensis, Candida spp., Sporothrix schenckii, Cryptococcus neoformans, Blastomyces dermatitidis, Coccidioides immitis, Aspergillus niger, and Aspergillus fumigatus were applied in the reaction. This rapid, sensitive, and specific assay provides a way to identify typical and atypical isolates of H. capsulatum var. capsulatum. PMID- 12574243 TI - New PCR assay using glucose-6-phosphate dehydrogenase for identification of Leishmania species. AB - Glucose-6-phosphate dehydrogenase (G6PD) is one of the multilocus enzymes used to identify Leishmania by zymodeme analysis. The polymorphic pattern revealed by partial characterization of the gene encoding G6PD generated molecular markers useful in the identification of different Leishmania species by PCR. Initially degenerate oligonucleotides were designed on the basis of data on the conserved active center described for other organisms. Primers for reverse transcription PCR experiments, designed from the nucleotide sequence of the PCR product, enabled us to characterize the 5' and 3' untranslated regions and the G6PD open reading frame of reference strains of Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, Leishmania (Leishmania) mexicana, and Leishmania (Leishmania) amazonensis. Sets of paired primers were designed and used in PCR assays to discriminate between the parasites responsible for tegumentar leishmaniasis of the subgenera Leishmania (Leishmania) and Leishmania (Viannia) and to distinguish L. (Viannia) braziliensis from others organisms of the subgenus Leishmania (Viannia). No amplification products were detected for the DNA of Crithidia fasciculata, Trypanosoma cruzi, or Leishmania (Sauroleishmania) tarentolae or DNA from a healthy human control. The tests proved to be specific and were sensitive enough to detect parasites in human biopsy specimens. The successful discrimination of L. (Viannia) braziliensis from other parasites of the subgenus Leishmania (Viannia) opens the way to epidemiological studies in areas where more than one species of the subgenus Leishmania (Viannia) exist, such as Amazonia, as well as follow-up studies after chemotherapy and assessment of clinical prognoses. PMID- 12574244 TI - Identification of a specific antigenic region of the P82 protein of Babesia equi and its potential use in serodiagnosis. AB - The efficacy of the Be82 gene product fused with glutathione S-transferase (GST/Be82) in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Babesia equi infection was reported previously (H. Hirata et al., J. Clin. Microbiol. 40:1470-1474, 2002). However, the ELISA with the GST/Be82 antigen cross-reacted with Babesia caballi-infected horse sera, despite the high rate of detection of B. equi. These results suggested that GST/Be82 has an antigen in common with B. caballi or antigenicity similar to that of B. caballi. In the present study, we constructed a series of five clones with deletions in the Be82 gene product, each of which was fused with GST, and used them in ELISAs in order to overcome the cross-reactivity seen with B. caballi. One of the deletion clones, a clone with a deletion of the Be82 gene from positions 236 to 381 (Be82/236-381), specifically and sensitively detected B. equi-infected horse sera without cross-reactivity with B. caballi-infected horse sera. Assays with clones from which other gene products were deleted showed decreased sensitivities or remained nonspecific for the detection of B. equi-infected horse sera. These results suggest that the Be82/236-381 gene product is a novel antigen for the diagnosis of B. equi infection in horses. PMID- 12574245 TI - Highly polymorphic microsatellite for identification of Candida albicans strains. AB - The polymorphism of a new microsatellite locus (CAI) was investigated in a total of 114 Candida albicans strains, including 73 independent clinical isolates, multiple isolates from the same patient, isolates from several episodes of recurrent vulvovaginal infections, and two reference strains. PCR genotyping was performed automatically, using a fluorescence-labeled primer, and in the 73 independent isolates, 26 alleles and 44 different genotypes were identified, resulting in a discriminatory power of 0.97. CAI was revealed to be species specific and showed a low mutation rate, since no amplification product was obtained when testing other pathogenic Candida species and no genotype differences were observed when testing over 300 generations. When applying this microsatellite to the identification of strains isolated from recurrent vulvovaginal infections in eight patients, it was found that 13 out of 15 episodes were due to the same strain. When multiple isolates, obtained from the same patient and plated simultaneously, were typed for CAI, the same genotype was found in each case, confirming that the infecting population was clonal. Moreover, the same genotype appeared in isolates from the rectum and the vagina, revealing that the former could be a reservoir of potentially pathogenic strains. This new microsatellite proves to be a valuable tool to differentiate C. albicans strains. Furthermore, when compared to other molecular genotyping techniques, CAI proved to be very simple, highly efficient, and reproducible, being suitable for low-quantity and very-degraded samples and for application in large-scale epidemiological studies. PMID- 12574246 TI - Diversity of bacterial populations on the tongue dorsa of patients with halitosis and healthy patients. AB - The primary purpose of the present study was to compare the microbial profiles of the tongue dorsa of healthy subjects and subjects with halitosis by using culture independent molecular methods. Our overall goal was to determine the bacterial diversity on the surface of the tongue dorsum as part of our ongoing efforts to identify all cultivable and not-yet-cultivated species of the oral cavity. Tongue dorsum scrapings were analyzed from healthy subjects with no complaints of halitosis and subjects with halitosis, defined as an organoleptic score of 2 or more and volatile sulfur compound levels greater than 200 ppb. 16S rRNA genes from DNA isolated from tongue dorsum scrapings were amplified by PCR with universally conserved bacterial primers and cloned into Escherichia coli. Typically, 50 to 100 clones were analyzed from each subject. Fifty-one strains isolated from the tongue dorsa of healthy subjects were also analyzed. Partial sequences of approximately 500 bases of cloned inserts from the 16S rRNA genes of isolates were compared with sequences of known species or phylotypes to determine species identity or closest relatives. Nearly complete sequences of about 1,500 bases were obtained for potentially novel species or phylotypes. In an analysis of approximately 750 clones, 92 different bacterial species were identified. About half of the clones were identified as phylotypes, of which 29 were novel to the tongue microbiota. Fifty-one of the 92 species or phylotypes were detected in more than one subject. Those species most associated with healthy subjects were Streptococcus salivarius, Rothia mucilaginosa, and an uncharacterized species of Eubacterium (strain FTB41). Streptococcus salivarius was the predominant species in healthy subjects, as it represented 12 to 40% of the total clones analyzed from each healthy subject. Overall, the predominant microbiota on the tongue dorsa of healthy subjects was different from that on the tongue dorsa of subjects with halitosis. Those species most associated with halitosis were Atopobium parvulum, a phylotype (clone BS095) of Dialister, Eubacterium sulci, a phylotype (clone DR034) of the uncultivated phylum TM7, Solobacterium moorei, and a phylotype (clone BW009) of STREPTOCOCCUS: On the basis of our ongoing efforts to obtain full 16S rRNA sequences for all cultivable and not-yet-cultivated species that colonize the oral cavity, there are now over 600 species. PMID- 12574247 TI - Serotyping of Listeria monocytogenes by enzyme-linked immunosorbent assay and identification of mixed-serotype cultures by colony immunoblotting. AB - Routine analysis of Listeria monocytogenes by serotyping using traditional agglutination methods is limited in use because of the expense and limited availability of commercially prepared antisera and intra- and interlaboratory discrepancies arising from differences in antiserum preparation and visual determination of agglutination. We have adapted a commercially available set of L. monocytogenes antisera to an enzyme-linked immunosorbent assay (ELISA) format for high-throughput, low-cost serotype determination. Rather than subjective visualization of agglutination, positive antigen and antiserum reactions were scored by a quantitative, colorimetric reaction. ELISA serotyping of 89 of 101 L. monocytogenes isolates agreed with slide agglutination serotyping data, and 100 previously uncharacterized isolates were serotyped unambiguously by the ELISA method. In addition, mixed-serotype cultures of L. monocytogenes were identified by a colony immunoblot procedure, in which serogroup 1/2 and serogroup 4 colonies were discriminated by differential staining. PMID- 12574248 TI - Quantitative assay of hepatitis C virus RNA using an automated extraction system for specific capture with probes and paramagnetic particle separation. AB - A commercially available automated specimen preparation instrument for specific probe capture and paramagnetic separation has been developed (AmpliCap/GT-12; Roche Molecular Systems). We evaluated assay performance of the AmpliCap/GT-12 in the quantitative assay for hepatitis C virus (HCV) RNA with the AMPLICOR HCV MONITOR Test (version 2.0). Assay linearity using serial dilutions from a serum panel was observed in the range of 500 to 850000 IU/ml, with a slightly compromised slope in the higher viral titers. The overall within-run and between run reproducibility of the entire detection process for 3 and 5 log(10) (IU/ml) of HCV RNA in samples had a standard deviation of <0.2, which was comparable to a manual method based on organic extraction and isopropanol precipitation (Roche Molecular Systems). Comparison of the test results with those obtained by the manual method showed a good correlation (R(2) = 0.972, n = 86). Using heparin (3, 6.5, and 13 U/ml), dextran sulfate (0.1, 1, and 5 mM), hemoglobin (1.13, 2.25, and 4.5 g/liter), conjugated or unconjugated bilirubin (7.5, 15, and 30 mg/dl), and ATP (1.25, 2.5, and 5.0 mM) as known inhibitors, inhibition was only detected at a dextran sulfate concentration of 1 mM with the manual method but not with the AmpliCap/GT-12 extraction. In summary, the AmpliCap/GT-12 system was shown to permit a stable extraction process and accurate results for the quantitative assay of HCV RNA, successfully eliminating the inhibitory effect of dextran sulfate. This automated extraction system provides reliable and reproducible test results and saves labor; thus, it is suitable for routine diagnostic PCR. PMID- 12574249 TI - Diagnosing herpesvirus infections by real-time amplification and rapid culture. AB - Procedures using real-time technique were developed to demonstrate the presence of herpes simplex virus type 1 (HSV-1) and HSV-2, varicella zoster virus (VZV), and cytomegalovirus (CMV) in miscellaneous clinical specimens. The assays were compared to rapid culture using centrifugation followed by detection with monoclonal antibodies. A total of 711 consecutive samples were collected from different patient groups. Throat swabs were obtained from transplant patients; dermal or oral specimens were collected from patients suspected for VZV or HSV infection. Genital specimens were taken from patients who attended the Clinic for Sexually Transmitted Diseases at the Dijkzigt Hospital Rotterdam presenting with symptoms of a primary genital ulcer. Nucleic acid extraction was carried out using a MagnaPure LC instrument. The amplification steps were performed on the ABI Prism 7700 sequence detection system. To monitor the process of extraction and amplification, a universal control consisting of seal herpesvirus type 1 (PhHV-1) was added to the clinical specimens. By culture 127 of 668 (19%) samples were positive for HSV-1, 72 of 668 (10.8%) specimens were positive for HSV-2, and 17 of 366 (4.6%) were positive for VZV. Using real-time amplification the numbers of positive specimens were 143 of 668 (21.4%), 97 of 668 (14.5%), and 27 of 366 (7.4%), respectively. Eighty-six specimens were tested for CMV, 12 (14.0%) were positive by culture, and 17 (19.8%) were positive by real-time PCR. The clinical data of the patients with discrepant results were reviewed thoroughly. In all cases the patients with only real-time PCR-positive results could be considered as truly infected. We concluded that the real-time amplification technique is suitable for the detection of human herpesvirus infection. It offers a semiquantitative and reliable assay with a quick result that is more sensitive than rapid culture, especially for the diagnosis of HSV-2 and VZV infections. PMID- 12574250 TI - Detection and identification of fungi from fungus balls of the maxillary sinus by molecular techniques. AB - The aim of this study was to find a reliable method for the detection and identification of fungi in fungus balls of the maxillary sinus and to evaluate the spectrum of fungi in these samples. One hundred twelve samples were obtained from patients with histologically proven fungal infections; 81 samples were paraffin-embedded tissue sections of the maxillary sinus. In 31 cases, sinus contents without paraffin embedding were sent for investigation. PCR amplification with universal fungal primers for 28S ribosomal DNA and amplicon identification by hybridization with species-specific probes for Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus, Aspergillus glaucus, Pseudallescheria boydii, Candida albicans, and Candida glabrata were performed for all samples. Furthermore, PCR products were sequenced. Fresh samples were also cultivated. Fungal DNA was detected in all of the fresh samples but only in 71 paraffin-embedded tissue samples (87.7%). Sequence analysis was the most sensitive technique, as results could be obtained for 28 (90.3%) fresh samples by this method in comparison to 24 (77.4%) samples by hybridization and 16 (51.6%) samples by culture. However, sequence analysis delivered a result for only 36 (50.7%) of the paraffin-embedded specimens. Hybridization showed reliable results for A. fumigatus, which proved to be the most common agent in fungus balls of the maxillary sinus. Other Aspergillus species and other genera were rarely found. PMID- 12574251 TI - Clonal diversity among streptogramin A-resistant Staphylococcus aureus isolates collected in French hospitals. AB - We analyzed 62 clinical isolates of streptogramin A-resistant (SGA(r)) Staphylococcus aureus collected between 1981 and 2001 in 14 hospitals located in seven French cities. These isolates, including five with decreased susceptibility to glycopeptides, were distributed into 45 antibiotypes and 38 SmaI genotypes. Each of these genotypes included between 1 and 11 isolates, the SmaI patterns of which differed by no more than three bands. Although numerous clones were identified, we observed the spread of monoclonal isolates either within the same hospital or within hospitals in distinct cities and at large time intervals. Hybridization with probes directed against 10 SGA(r) genes (vatA, vatB, vatC, vatD, vatE, vgaA, vgaB, vgaAv, vgbA, and vgbB) revealed six patterns: vgaAv (21 isolates), vatA-vgbA (24 isolates), vgaAv-vatB-vgaB (14 isolates), vgaAv-vatA vgbA (1 isolate), vgaAv-vatA-vgbA-vatB-vgaB (1 isolate), and vgaA (1 isolate). We detected at least one SGA(r) determinant in all of the tested isolates. vgaAv, which is part of the recently characterized transposon Tn5406, was found in 59.7% of the tested isolates. Of the 16 streptogramin B-susceptible isolates, 14 carried vgaAv alone and were susceptible to the mixtures of streptogramins, whereas the 2 isolates carrying vgaAv-vatB-vgaB were resistant to these mixtures. vatA-vgbA was found on plasmids of the same apparent size in 26 (42%) of the tested clinical isolates from 18 unrelated SmaI genotypes. The possible dissemination of some of the multiple clones characterized in the present study with an expected increased selective pressure of streptogramins following the recent licensing of Synercid (quinupristin-dalfopristin) must be carefully monitored. PMID- 12574252 TI - Comparison of a new quantitative ompA-based real-Time PCR TaqMan assay for detection of Chlamydia pneumoniae DNA in respiratory specimens with four conventional PCR assays. AB - Chlamydia pneumoniae, an important respiratory pathogen, is difficult to culture, and detection rates by conventional PCRs vary considerably. A new quantitative ompA-based real-time PCR assay based on TaqMan technology for detection of C. pneumoniae in respiratory samples is described, and its performance in terms of sensitivity and reproducibility is compared with those of four published conventional PCRs (one single-step PCR targeting a cloned PstI fragment; two nested PCRs, one targeting the 16S rRNA gene followed by hybridization and the other targeting the ompA gene; and a touchdown enzyme time-release [TETR] PCR also targeting the 16S rRNA gene). Both ompA-based PCRs showed the best analytical sensitivity. All five assays could detect even lower target levels from spiked sputum, with the 16S rRNA assays performing better than the ompA based nested PCR (10(-6) inclusion-forming units [IFU] were detected in four of four and two of four replicates by the 16S rRNA TETR PCR and the 16S rRNA nested PCR, respectively). In general, the ompA-based real-time protocol produced the most consistent positive results for all replicates tested down to 10(-6) IFU. Eight of 45 patient sputum specimens (18%) were C. pneumoniae DNA positive in at least one of four replicates tested by at least one assay. Without taking into consideration the analytical sensitivity or the reproducibility of the test results, the numbers of C. pneumoniae DNA-positive sputum specimens (n = 8) were four, three, two, two, and one for the 16S rRNA TETR assay, the PstI-based single step PCR, the ompA-based real-time PCR, the ompA-based nested touchdown PCR, and the 16S rRNA-based nested PCR, respectively. However, the overall rate of concordance of positive results was low. Only one cell culture-positive sputum specimen was positive by four of five assays (14 of 16 replicates; mean cycle threshold value, 25; 10(8) particles/ml of sputum). Thirty-seven specimens were C. pneumoniae negative by all five assays for all replicates tested, as were all negative controls (n = 65 to 100 per testing panel). No PCR inhibitors were detected by real-time PCR or by the 16S rRNA-based nested assay. We confirm that the analytical sensitivity of an assay for the detection of C. pneumoniae does not necessarily predict its ability to detect its target in sputum. A quantitative, fast, and easy-to-handle diagnostic approach such as the ompA-based real-time TaqMan PCR described here might improve the detection of C. pneumoniae in respiratory samples. PMID- 12574253 TI - Evaluation of semiautomated multiplex PCR assay for determination of Streptococcus pneumoniae serotypes and serogroups. AB - A semiautomated method for the determination of five serotypes and three serogroups in Streptococcus pneumoniae was developed. Primers specific for serotypes 1, 3, 14, 19F, and 23F and serogroups 6, 19, and 23 were combined in three multiplex PCRs. Products were separated by capillary electrophoresis with a 7-min run time, and a serotype or serogroup was assigned on the basis of fragment size. The method was used to test 93 clinical isolates, and all isolates of the serotypes concerned were correctly detected. The strategy would allow the detection of multiple serotypes in a single sample. Detection of additional serotypes could be included as capsule locus sequences become available. PMID- 12574254 TI - Human papillomavirus type 16 status in cervical carcinoma cell DNA assayed by multiplex PCR. AB - Integration of human papillomavirus (HPV) DNA into host genome occurs early in cancer development and is probably an important event in malignant transformation of cervical cancer. The HPV genome integration usually disrupts E2 gene open reading frames. It results in the lack of E2 gene suppressor of the synthesis of E6 and E7 products which, in turn, leads to the overexpression of E6 and E7 genes. The oncogenic HPV types (HPV16, -18, -45, and -58) can be present as episomes or may integrate into human chromosomes. Sixty-six cervical cancer patients positive for HPV16 were tested for the presence of E6, E2, E1, and L1 genes. Multiplex PCR was carried out in all cases. Using cluster analysis, the calculated ratios of E1/E6, E2/E6, L1/E6, E1/E2, and E2/(E1*E6) gene amplification products were divided into two or three statistically different groups. These were used for statistical analysis of the prevalence of specific gene types in histological types of cancer, different levels of clinical staging, and histologically confirmed nodal metastases. The statistical analysis proved a significant correlation in the ratios of E2/E6 and E1/E2 only. The E2/E6 and E1/E2 were higher in carcinoma in situ than in advanced squamous cancers. The E2/E6 ratios were lower in higher clinical stages. The multiplex PCR estimation of the E2/E6 ratio could be a simple method for selecting patients with a high risk of a poor outcome in a standard stage-dependent treatment procedure. PMID- 12574255 TI - Analysis of a viridans group strain reveals a case of bacteremia due to lancefield group G alpha-hemolytic Streptococcus dysgalactiae subsp equisimilis in a patient with pyomyositis and reactive arthritis. AB - Streptococcus dysgalactiae is classified by a combination of phenotypic and genotypic characteristics into Lancefield group C alpha-hemolytic Streptococcus dysgalactiae subsp. dysgalactiae and Lancefield group C, group G, and group L beta-hemolytic Streptococcus dysgalactiae subsp. equisimilis. In this study, we report the isolation of a catalase-negative, alpha-hemolytic, optochin- and bacitracin-resistant viridans group strain, which does not grow in 10 or 40% bile, on MacConkey agar or bile esculin agar, or in 6% NaCl, from the blood culture of a 73-year-old woman with pyomyositis and poststreptococcal reactive arthritis. Lancefield grouping revealed that the strain was a group G streptococcus. The Vitek system (GPI) showed that it was unidentified, and the API system (20 STREP) showed that it was 95.7% S. dysgalactiae subsp. dysgalactiae. 16S rRNA gene sequencing showed that it was a strain of S. dysgalactiae. Based on phylogenetic affiliation with 16S rRNA gene or GroEL amino acid (another bacterial gene, in addition to 16S rRNA gene, that is highly conserved) sequences, the strain is most closely related to Lancefield group C beta-hemolytic S. dysgalactiae subsp. equisimilis. PCR amplification and sequencing of the streptolysin S structural gene (sagA) and M protein gene (emm) hypervariable region showed the presence of these suspected primary virulence factors. Further studies would delineate whether the isolate is just a hemolysin deficient variant of group G beta-hemolytic S. dysgalactiae subsp. equisimilis or a novel type of S. dysgalactiae. The present case showed that group G alpha hemolytic S. dysgalactiae subsp. equisimilis can be associated with serious invasive infection and poststreptococcal sequelae. PMID- 12574256 TI - Multiple patterns of resistance to fluconazole in Candida glabrata isolates from a patient with oropharyngeal candidiasis receiving head and neck radiation. AB - Candida glabrata has emerged in recent years as a significant cause of systemic fungal infection. We have previously reported on the first three patients receiving radiation for head and neck cancer to develop oropharyngeal candidiasis due to C. glabrata. The goal of this study was to track the development of increased fluconazole resistance in C. glabrata isolates and to evaluate previously described genetic mechanisms associated with this resistance from one of these three patients. The patient was a 52-year-old man with squamous cell carcinoma treated with radiation. At week 7 of his radiation, he developed oropharyngeal candidiasis, which was treated with 200 mg of fluconazole daily for 2 weeks. Serial cultures from this and three subsequent time points yielded C. glabrata. Isolates from these cultures were subjected to antifungal susceptibility testing, DNA karyotyping, and evaluation of the expression of genes previously associated with C. glabrata resistance to fluconazole, CgCDR1, CgCDR2, and CgERG11. Two strains (A and B) of C. glabrata were identified and found to display different patterns of resistance development and gene expression. Strain A developed resistance over a 2-week period and showed no overexpression of these genes. In contrast, strain B first showed resistance 6 weeks after fluconazole therapy was discontinued but showed overexpression of all three genes. In conclusion, development of resistance to fluconazole by C. glabrata is a highly varied process involving multiple molecular mechanisms. PMID- 12574257 TI - Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens. AB - There is an increasing demand for diagnostic testing for Giardia intestinalis (G. lamblia) and Cryptosporidium parvum, with a priority being placed on obtaining diagnostic results in an efficient and timely manner. Several commercial companies have developed rapid diagnostic tests that are simple to perform and can be completed in less time than traditional methods for detecting Giardia and Cryptosporidium: We compared one of these rapid tests, the ImmunoCard STAT! (Meridian Bioscience, Inc.) lateral-flow immunoassay, with the MERIFLUOR direct fluorescent-antibody (DFA) test, the ProSpecT EZ microplate assay for Giardia and the ProSpecT microplate assay for Cryptosporidium, and modified Kinyoun's acid fast stained smears for the detection of Cryptosporidium using 246 specimens. The MERIFLUOR DFA (Meridian Bioscience, Inc.) test detected the largest number of cases (32 Giardia and 37 Cryptosporidium) infections and was used to calculate the sensitivity and specificity of the other tests. For Giardia, the sensitivities of the ImmunoCard STAT! and the ProSpecT Giardia EZ microplate assay (Alexon-Trend, Inc.) were 81 and 91%, respectively. For detection of Cryptosporidium, the sensitivities of the ImmunoCard STAT!, the ProSpecT Cryptosporidium microplate assay (Alexon-Trend, Inc.), and modified Kinyoun's acid-fast stained smears were 68, 70, and 78%, respectively. Test specificities were equal to or greater than 99%. Specimens with very small numbers of organisms were not detected by the ImmunoCard STAT!, the ProSpecT microplate assay or modified Kinyoun's acid-fast stained smears. PMID- 12574258 TI - Multisite reproducibility of results obtained by two broth dilution methods for susceptibility testing of Mycobacterium avium complex. AB - A multicenter study was conducted to assess the interlaboratory reproducibility of susceptibility testing of Mycobacterium avium complex (MAC) by broth microdilution using two different media (cation-adjusted Mueller-Hinton broth with 5% oleic acid-albumin-dextrose-catalase and 7H9 broth with casein) and by macrodilution using the BACTEC 460TB and 12B media at pH 6.8 and 7.3 to 7.4. Ten well-characterized strains of MAC (four macrolide susceptible, six macrolide resistant) were tested against clarithromycin and azithromycin (the latter only by BACTEC 460TB, pH 6.8). At each site, strains were tested against clarithromycin three times on each of three separate days (nine testing events per isolate) by using a common lot of microdilution trays and BACTEC 12B medium, pH 6.8; strains were tested once on three separate days against clarithromycin in 12B medium at pH 7.3 to 7.4 and against azithromycin. Agreement among MICs (i.e., mode +/- 1 twofold dilution) was 100% for all strains and both drugs when BACTEC 460TB was used, regardless of the pH of the medium, but varied when microdilution with either medium was used, particularly with susceptible strains. Agreement based on interpretive category, with NCCLS-recommended breakpoints, was 100% for all strains with the BACTEC 460TB method (both drugs and both pH values) and with microdilution using 7H9 broth. With microdilution and Mueller-Hinton broth, agreement by interpretive category was 100% for eight isolates and >90% for two; errors occurred only in laboratories where personnel had minimal experience with this technique. MAC susceptibility testing may be performed by broth macrodilution or microdilution at either pH, with NCCLS-recommended interpretive breakpoints. However, because visual interpretation of broth microdilution end points is subjective, it is more prone to reader error; therefore, this method requires greater expertise than the BACTEC 460TB. Both techniques require appropriate validation and continued documentation of proficiency. PMID- 12574259 TI - Mixed-genome microarrays reveal multiple serotype and lineage-specific differences among strains of Listeria monocytogenes. AB - Epidemiological studies and analysis of putative virulence genes have shown that Listeria monocytogenes has diverged into several phylogenetic divisions. We hypothesize that similar divergence has occurred for many genes that influence niche-specific fitness and virulence and that identifying these differences may offer new opportunities for the detection, treatment, and control of this important pathogen. To explore this issue further, we developed a microarray composed of fragmented DNA taken from 10 strains of L. monocytogenes. We then hybridized genomic DNA from 50 different strains to replicate arrays and analyzed the resulting hybridization patterns. A simple Euclidean distance metric permitted the reconstruction of previously described genetic relationships between serotypes, and only four microarray probes were needed to discriminate between the most important serotypes (1/2a, 1/2b, 1/2c, and 4). We calculated an index of linkage equilibrium from the microarray data and confirmed that L. monocytogenes has a strongly clonal population structure (I(A) = 3.85). Twenty nine informative probes were retrieved from the library and sequenced. These included genes associated with repairing UV-damaged DNA, salt tolerance, biofilm formation, heavy metal transport, ferrous iron transport, and teichoic acid synthesis. Several membrane-bound lipoproteins and one internalin were identified, plus three phage sequences and six sequences with unknown function. Collectively, these data confirm that many genes have diverged between lineages of L. monocytogenes. Furthermore, these results demonstrate the value of mixed genome microarrays as a tool for deriving biologically useful information and for identifying and screening genetic markers for clinically important microbes. PMID- 12574260 TI - Characterization of some actinomyces-like isolates from human clinical sources: description of Varibaculum cambriensis gen nov, sp nov. AB - Fifteen strains of an anaerobic, catalase-negative, gram-positive diphtheroid shaped bacterium recovered from human sources were characterized by phenotypic and molecular chemical and molecular genetic methods. The unidentified bacterium showed some resemblance to Actinomyces species and related taxa, but biochemical testing, polyacrylamide gel electrophoresis analysis of whole-cell proteins, and amplified 16S ribosomal DNA restriction analysis indicated the strains were distinct from all currently named Actinomyces species and related taxa. Comparative 16S rRNA gene sequencing studies showed that the bacterium represents a hitherto-unknown phylogenetic line that is related to but distinct from Actinomyces, Actinobaculum, Arcanobacterium, and Mobiluncus: We propose, on the basis of phenotypic and phylogenetic evidence, that the unknown bacterium from human clinical specimens should be classified as a new genus and species, Varibaculum cambriensis gen. nov., sp. nov. The type strain of Varibaculum cambriensis sp. nov. is CCUG 44998(T) = CIP 107344(T). PMID- 12574261 TI - Bartonella strains from ground squirrels are identical to Bartonella washoensis isolated from a human patient. AB - The most likely animal source of a human case of cardiac disease in Washoe County, Nev., was identified by comparison of DNA sequences of three genes (citrate synthase gltA, 60-kDa heat shock protein gene groEL, and 16S rRNA gene) of Bartonella washoensis cultured from the human patient in question and of Bartonella isolates obtained from the following Nevada rodents: Peromyscus maniculatus (17 isolates), Tamias minimus (11 isolates), Spermophilus lateralis (3 isolates), and Spermophilus beecheyi (7 isolates). Sequence analyses of gltA amplicons obtained from Bartonella from the rodents demonstrated considerable heterogeneity and resulted in the identification of 16 genetic variants that were clustered within three groups in phylogenetic analysis. Each of the three groups was associated with a rodent genus, Peromyscus, Tamias, or Spermophilus: The gltA, 16S rRNA gene, and groEL sequences of a Bartonella isolate obtained from a California ground squirrel (S. beecheyi) were completely identical to homologous sequences of B. washoensis, strongly suggesting that these animals were the source of infection in the human case. PMID- 12574262 TI - Multicenter evaluation of the VERSANT HCV RNA qualitative assay for detection of hepatitis C virus RNA. AB - We report the results of a multicenter evaluation of a new assay for the detection of hepatitis C virus (HCV) RNA in human serum or plasma based on transcription-mediated amplification (HCV TMA). Analysis of combined data obtained from 15 independent sites, including 4 sites in the United States and 11 in Europe, by using preproduction kits showed a limit of detection of 9.8 IU/ml and an overall mean specificity of 97.9%. In addition, assay runs and samples were valid consistently (97.8% of assay runs and 98.0% of specimen results). Of the 15 sites that participated in the multicenter evaluation, 2 subsequently carried out additional performance studies with production kits in support of the assay's registration in France. Comparison of the findings from these two sites during the multicenter evaluation and during the registration studies showed an overall improvement in assay performance. A statistically significant (P < 0.001) improvement was achieved for both specificity and specimen validity in the registration studies, which were 99.4 and 98.1%, respectively. Combined data from the two sites showed a lower limit of detection of approximately 2.4 IU/ml and an improved assay validity of 100%, although the sample size was too small to show statistical significance at the 0.05 level. In summary, the performance characteristics of HCV TMA indicate that this assay is a reliable tool for the detection of HCV RNA in serum or plasma. Improvement in assay performance has been demonstrated with refinement of assay reagents, instrumentation, and operator experience. PMID- 12574263 TI - Identification of major Streptococcal species by rrn-amplified ribosomal DNA restriction analysis. AB - Amplified ribosomal DNA restriction analysis (rrn-ARDRA) is based on PCR amplification and restriction of a fragment of rRNA genes including 16S and 23S genes and the intergenic spacer. rrn-ARDRA was evaluated for the identification of species within the genus Streptococcus: A total of 148 type and reference strains of pyogenic, oral, and group D streptococci were examined in order to construct a database for identification of streptococci. The amplified product was a single band approximately 4500 bp long. This amplicon was digested separately with three (HhaI, MboII, and Sau3A) restriction endonucleases. Respectively, 27, 26, and 28 major patterns were observed after HhaI, MboII, and Sau3A restrictions. Streptococcal strains belonging to different species had different patterns or different combination of patterns. An identification system based upon a combination of the three restriction patterns in a single database was then proposed. rrn-ARDRA was successfully applied to 11 clinical isolates whose identification to the species level was difficult to obtain by phenotypic analysis. Using a database of well-characterized strains, rrn-ARDRA is a powerful method for the identification of streptococcal isolates. PMID- 12574264 TI - Six rapid tests for direct detection of Clostridium difficile and its toxins in fecal samples compared with the fibroblast cytotoxicity assay. AB - Clostridium difficile is one of the most frequent causes of nosocomial gastrointestinal disease. Risk factors include prior antibiotic therapy, bowel surgery, and the immunocompromised state. Direct fecal analysis for C. difficile toxin B by tissue culture cytotoxin B assay (CBA), while only 60 to 85% sensitive overall, is a common laboratory method. We have used 1,003 consecutive, nonduplicate fecal samples to compare six commercially available immunoassays (IA) for C. difficile detection with CBA: Prima System Clostridium difficile Tox A and VIDAS Clostridium difficile Tox A II, which detect C. difficile toxin A; Premier Cytoclone A/B and Techlab Clostridium difficile Tox A/B, which detect toxins A and B; and ImmunoCard Clostridium difficile and Triage Micro C. difficile panels, which detect toxin A and a species-specific antigen. For all tests, Triage antigen was most sensitive (89.1%; negative predictive value [NPV] = 98.7%) while ImmunoCard was most specific (99.7%; positive predictive value [PPV] = 95.0%). For toxin tests only, Prima System had the highest sensitivity (82.2%; NPV = 98.0%) while ImmunoCard had the highest specificity (99.7%; PPV = 95.0%). Hematopoietic stem cell transplant (HSCT) patients contributed 44.7% of all samples tested, and no significant differences in sensitivity or specificity were noted between HSCT and non-HSCT patients. IAs, while not as sensitive as direct fecal CBA, produce reasonable predictive values, especially when both antigen and toxin are detected. They also offer significant advantages over CBA in terms of turnaround time and ease of use. PMID- 12574265 TI - Longitudinal reliability of focus glycoprotein G-based type-specific enzyme immunoassays for detection of herpes simplex virus types 1 and 2 in women. AB - Serologic assays that utilize herpes simplex virus (HSV) type-specific glycoproteins G-1 (HSV-1) and G-2 (HSV-2) to discriminate between antibodies against HSV-1 and HSV-2 are sensitive and specific. However, the high rates of seroreversion, defined as the change in an individual's antibody status from positive to negative over time, previously reported in longitudinal evaluations of glycoprotein G type-specific tests suggests that their use in HSV acquisitional studies would be problematic. To further explore the reliability of the glycoprotein G-based serologic tests, we evaluated HSV-1 and HSV-2 enzyme immunoassays from Focus Technologies in a longitudinal cohort of 1207 young women from Pittsburgh, Pa. On enrollment of the women in the study, HSV-1 and HSV-2 antibodies were detected in 46.6 and 24.9% of the women, respectively. Among the women with at least three visits, 3.4% (15 of 447) of those who were HSV-1 antibody positive had a subsequent negative result while fewer than 1% (2 of 227) of those who were HSV-2 antibody positive seroreverted. The median of mean positive index values for women who seroreverted to HSV-1 antibody was lower than that for women who remained seropositive (1.25 versus 7.06; P < 0.001). Similarly, the median of mean positive index values for women whose HSV-2 antibody status reverted from positive to negative was lower than that for those women who did not serorevert (1.83 versus 7.46; P = 0.02). Comparative Western blot analysis demonstrated that the lower positive index values, seen more often among the HSV seroreverters, often signified false-positive immunoassay results. Overall, the seroreversion rates were low; the use of glycoprotein G-based serologic tests for the measurement of HSV-1 and HSV-2 antibodies in incidence studies therefore appears warranted. PMID- 12574266 TI - Multilocus sequence typing reveals a lack of diversity among Escherichia coli O157:H7 isolates that are distinct by pulsed-field gel electrophoresis. AB - Escherichia coli O157:H7 is a major cause of foodborne illness in the United States. Pulsed-field gel electrophoresis (PFGE) is the molecular epidemiologic method mostly commonly used to identify food-borne outbreaks. Although PFGE is a powerful epidemiologic tool, it has disadvantages that make a DNA sequence-based approach potentially attractive. Multilocus sequence typing (MLST) analyzes the internal fragments of housekeeping genes to establish genetic relatedness between isolates. We sequenced selected portions of seven housekeeping genes and two membrane protein genes (ompA and espA) of 77 isolates that were diverse by PFGE to determine whether there was sufficient sequence variation to be useful as an epidemiologic tool. There was no DNA sequence diversity in the sequenced portions of the seven housekeeping genes and espA. For ompA, all but five isolates had sequence identical to that of the reference strains. E. coli O157:H7 has a striking lack of genetic diversity in the genes we explored, even among isolates that are clearly distinct by PFGE. Other approaches to identify improved molecular subtyping methods for E. coli 0157:H7 are needed. PMID- 12574267 TI - Use of a phage-based assay for phenotypic detection of mycobacteria directly from sputum. AB - The phage amplified biologically assay is a new method for rapid and low-cost phenotypic determination of the drug sensitivities of Mycobacterium tuberculosis isolates and the detection of viable organisms in patient specimens. Infection of slowly growing mycobacteria with phage (phage D29) was followed by chemical virucide destruction of extracellular phage. Infected mycobacteria were mixed in culture with rapidly growing sensor cells, which the phage can also infect; i.e., lytic amplification of phage occurs. The aims of the present study were to optimize the speed and sensitivity of the assay and reduce its cost for developing countries by using an M. tuberculosis-spiked sputum model with (i). identification of inhibitory components of sputum and optimization of decontamination methods; (ii). simplification of the washing and development steps; (iii). reduction of the use of high-cost components, e.g., oleate-albumin dextrose-catalase (OADC) supplement; and (iv). optimization of virucide treatment. The following results were obtained. (i). An inhibitory factor in sputum which could be removed by treatment of the sample with sodium dodecyl sulfate or NaOH decontamination was identified. (ii). A microcentrifuge-based approach with thixotropic silica as a bedding and resuspension agent was developed as an alternative to conventional centrifugation medium exchange. The yield was increased 228-fold, with increased speed and reduced cost. (iii). At present, after extracellular inactivation of phage, the ferrous ammonium sulfate (FAS) virucide is sequestered by dilution with an expensive supplement, OADC. Sodium citrate with calcium chloride was found to be a cost-effective after treatment with the FAS protectant and offered greater protection than OADC. Kinetic-lysis experiments indicated that an infection time of 1 to 3 h prior to FAS addition was optimal. (iv). Amplification of the signal (which corresponded to the burst size) was shown by allowing lysis prior to plating in a spiked medium model (up to 20-fold) and a spiked sputum model (up to 10-fold). A liquid culture detection method capable of detecting approximately 60 viable M. tuberculosis organisms in 1 ml of sputum was developed. Taken together, these improvements support the routine application of the assay to sputum specimens. PMID- 12574268 TI - Detection of Francisella tularensis within infected mouse tissues by using a hand held PCR thermocycler. AB - The diagnosis of human cases of tularemia often relies upon the demonstration of an antibody response to Francisella tularensis or the direct culturing of the bacteria from the patient. Antibody response is not detectable until 2 weeks or more after infection, and culturing requires special media and suspicion of tularemia. In addition, handling live Francisella poses a risk to laboratory personnel due to the highly infectious nature of this pathogen. In an effort to develop a rapid diagnostic assay for tularemia, we investigated the use of TaqMan 5' hydrolysis fluorogenic PCR to detect the organism in tissues of infected mice. Mice were infected to produce respiratory tularemia. The fopA and tul4 genes of F. tularensis were amplified from infected spleen, lung, liver, and kidney tissues sampled over a 5-day period. The samples were analyzed using the laboratory-based Applied Biosystems International 7900 and the Smiths Detection Edgewood BioSeeq, a hand-held portable fluorescence thermocycler designed for use in the field. A comparison of culturing and PCR for detection of bacteria in infected tissues shows that culturing was more sensitive than PCR. However, the results for culture take 72 h, whereas PCR results were available within 4 h. PCR was able to detect infection in all the tissues tested. Lung tissue showed the earliest response at 2 days when tested with the ABI 7900 and in 3 days when tested with the BioSeeq. The results were in agreement between the ABI 7900 and the BioSeeq when presented with the same sample. Template preparation may account for the loss of sensitivity compared to culturing techniques. The hand-held BioSeeq thermocycler shows promise as an expedient means of forward diagnosis of infection in the field. PMID- 12574269 TI - Molecular characterization and drug resistance patterns of strains of Mycobacterium tuberculosis isolated from patients in an AIDS counseling center in Port-au-Prince, Haiti: a 1-year study. AB - Tuberculosis (TB) is one of the most common opportunistic diseases that appear among human immunodeficiency virus (HIV)-positive patients in Haiti. In this context the probable emergence of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis is of great epidemiological concern. However, as routine culture of M. tuberculosis and drug susceptibility testing are not performed in Haiti, it has not been possible so far to evaluate the rate of drug resistance among M. tuberculosis isolates from circulating TB cases. This report describes the first study on the molecular typing and drug resistance of M. tuberculosis isolates from patients with culture-positive pulmonary tuberculosis monitored at the GHESKIO Centers in Haiti during the year 2000. Clinical, epidemiological, and drug susceptibility testing results were available for 157 patients with confirmed cases of TB, with a total of 8.9% of patients harboring MDR M. tuberculosis. A significant association between the occurrence of resistance and previous TB treatment was observed (P < 0.001), suggesting that a previous history of TB treatment was a risk factor associated with MDR TB in Haiti. The DNAs of individual isolates from 106 samples were available and were typed by spoligotyping and determination of the variable number of tandem DNA repeats. Both typing methods provided interpretable results for 96 isolates, and the clusters observed were further confirmed by ligation-mediated PCR to define potential cases of active transmission. Thirty-three (34%) of the isolates were found to be grouped into 11 clusters with two or more identical patterns. However, an assessment of risk factors (sex, HIV positivity, previous treatment, drug resistance) showed that none was significantly associated with the active transmission of TB. These observations suggest that acquired MDR TB is prevalent in Haiti and may be associated with compliance issues during TB treatment since prior TB therapy is the strongest risk factor associated with MDR TB. Prevention of TB transmission in Haiti should target active case investigation, routine detection of drug resistance, and adequate treatment of patients. The use of directly observed short-course therapy should be enforced throughout the country; and relapses, reactivations, or newly acquired infections should be discriminated by genotyping methods. PMID- 12574270 TI - Clonal reproduction and limited dispersal in an environmental population of Cryptococcus neoformans var gattii isolates from Australia. AB - Cryptococcus neoformans var. gattii is a causative agent of cryptococcosis and is thought to have a specific ecological association with a number of Eucalyptus species in Australia. However, the role that the tree plays in the life cycle of the fungus and the nature of the infectious propagule are not well understood. This study set out to examine whether sexual recombination is occurring in a natural population of C. neoformans var. gattii and whether the fungus disseminates between colonized trees. Thirty C. neoformans var. gattii isolates, consisting of both the alpha and a mating types, were collected from 13 Eucalyptus camaldulensis trees growing along a riverbank in Renmark, South Australia. The genetic diversity within the population was studied by using amplified fragment length polymorphism fingerprinting, and each isolate was assigned a unique multilocus genotype. Population genetic analyses of the multilocus data found no evidence of genetic exchange between members of the population, indicating a clonal population structure. Canonical variate analysis was then used to study the relationship between isolates from different colonized trees. Isolates from individual trees were strongly correlated, and it appeared that dispersal between trees was not occurring to any appreciable extent. These results suggest that the eucalypt may not be the primary niche for C. neoformans var. gattii but that the decaying wood present in hollows on these trees may provide a favorable substrate for extensive clonal propagation of the yeast cells. PMID- 12574271 TI - Simple method for screening Candida species isolates for the presence of secreted proteinases: a tool for the prediction of successful inhibitory treatment. AB - The yeasts of the genus Candida are opportunistic pathogens associated with the rising incidence of life-threatening infections in immunocompromised individuals. Secretion of aspartic proteinases has been determined to be one of the virulence factors of the pathogenic Candida species. To analyze the extracellular proteolytic activities of a large number of Candida clinical isolates, we developed a screening system based on a solid medium containing hemoglobin as the sole nitrogen source. The cleavage of hemoglobin by the secreted proteinases results in formation of clearance zones. The visibility of such zones was enhanced by addition of an acid-base indicator. Using this system, we assessed 245 clinical isolates of Candida from patients in the hospital of the Faculty of Medicine, Palacky University, Olomouc, Czech Republic, for the presence of secreted aspartic proteases (Saps). We also used the test plates for rapid semiquantitative testing of Sap inhibitors. Most of the pepstatin analogs affected the formation of the zones of clearance as well as the growth of Candida albicans, C. tropicalis, and C. parapsilosis colonies. By contrast, the human immunodeficiency virus proteinase inhibitors saquinavir, ritonavir, nelfinavir, and indinavir had no effect on the Candida strains tested. These results are in agreement with the inhibition constants obtained for the individual inhibitors with purified Saps. Thus, the plates containing hemoglobin proved to be an appropriate tool for the rapid and reliable assessment of Sap production and inhibition. PMID- 12574272 TI - Comparison of PCR assays for detection of the agent of human granulocytic ehrlichiosis, Anaplasma phagocytophilum. AB - Human granulocytic ehrlichiosis is an emerging infectious disease in the United States and Europe, and PCR methods have been shown to be effective for the diagnosis of acute infections. Numerous PCR assays and primer sets have been reported in the literature. The analytical sensitivities (limits of detection) of 13 published PCR primer sets were compared using DNA extracted from serial dilutions of Anaplasma phagocytophilum-infected HL-60 cells. The specificity of the assays that were able to detect 7 days were performed. Potential environmental reservoirs and possible risk factors for acquisition of C. tropicalis were searched for. Molecular epidemiologic studies by two methods of restriction fragment length polymorphism analysis and two methods of random amplified polymorphic DNA analysis were performed. Seventy-two neonates were colonized by yeasts (12.1%), of which 30 were colonized by Candida albicans, 17 were colonized by C. tropicalis, and 5 were colonized by Candida parapsilosis. From December 1998 to December 1999, 10 cases of fungemia occurred; 6 were due to C. parapsilosis, 2 were due to C. tropicalis, 1 was due to Candida glabrata, and 1 was due to Trichosporon asahii (12.8/1,000 admissions). Fungemia occurred more frequently in colonized than in noncolonized neonates (P < 0.0001). Genetic analysis of 11 colonization isolates and the two late blood isolates of C. tropicalis demonstrated two genotypes. One blood isolate and nine colonization isolates belonged to a single type. The fungemia/colonization ratio of C. parapsilosis (3/5) was greater than that of C. tropicalis (2/17, P = 0.05), other non-C. albicans Candida spp. (1/11, P = 0.02), or C. albicans (0/27, P = 0.05). Extensive environmental cultures revealed no common source of C. tropicalis or C. parapsilosis. There was neither prophylactic use of azoles nor other risk factors found for acquisition of C. tropicalis except for total parenteral nutrition. A substantial risk of colonization by non-C. albicans Candida spp. in the neonatal ICU may lead to a preponderance of C. tropicalis as a significant cause of neonatal fungemia. PMID- 12574277 TI - Molecular fingerprinting of fish-pathogenic Lactococcus garvieae strains by random amplified polymorphic DNA analysis. AB - In this work, we used the random amplified polymorphic DNA (RAPD) technique to evaluate the genetic diversity in Lactococcus garvieae, an important pathogen for fish. Fifty-seven strains with different hosts and geographical origins, including Japan and several countries of the Mediterranean area such as Spain, Portugal, France, Italy, England, and Turkey, were analyzed. Two primers, oligonucleotides 5 and 6 (Pharmacia Biotech) were utilized; primer 5 was the most discriminative, since allowed us to differentiate 10 RAPD -types related to the origin of the strains. Regardless of the oligonucleotide primer employed, the 57 isolates of L. garvieae studied were separated into three genetic groups, composed of the Spanish, Portuguese, English, and Turkish strains (group A), the Italian and French strains (group B), and the Japanese strains (group C). The similarity of isolates within each group, estimated on the basis of the Dice coefficient, ranged from 75 to 100%. Our findings also indicate that RAPD profiling constitutes a useful tool for epidemiological studies of this fish pathogen. PMID- 12574278 TI - Development of a multilocus sequence typing method for analysis of Listeria monocytogenes clones. AB - This study is a first step in the development of multilocus sequence typing (MLST) method for Listeria monocytogenes. Nine housekeeping genes were analyzed in a set of 62 strains isolated from different sources and geographic locations in Spain. These strains were previously characterized by pulsed-field gel electrophoresis (PFGE). Because of low diversity, two loci were discarded from the study. The sequence analysis of the seven remaining genes showed 29 different allelic combinations, with 22 of them represented by only one strain. The results of this sequence analysis were generally consistent with those of PFGE. Because MLST allows the easy comparison and exchange of results obtained in different laboratories, the future application of this new molecular method could be a useful tool for the listeriosis surveillance systems that will allow the identification and distribution of analysis of L. monocytogenes clones in the environment. PMID- 12574279 TI - Molecular diagnosis of infective endocarditis by PCR amplification and direct sequencing of DNA from valve tissue. AB - We used broad-range eubacterial PCR amplification followed by direct sequencing to identify microbial pathogens in heart valve material from 29 patients with histologically confirmed infective endocarditis and 23 patients free of infective endocarditis. Microorganisms cultured by conventional techniques matched those identified by PCR in 21 cases. PCR alone identified the causative agent in three cases (Streptococcus bovis, Staphylococcus cohnii, and Coxiella burnetii), allowing better patient management. PCR corrected the initial bacteriological diagnosis in three cases (Streptococcus bovis, Streptococcus mutans, and Bartonella henselae). Among the 29 cases of histologically confirmed infective endocarditis, PCR findings were positive in 27 cases and were consistent with the bacterial morphology seen at Gram staining (26 cases) or with the results obtained by immunohistologic analysis with an anti-C. burnetii monoclonal antibody (one case). In two other cases of histologically confirmed infective endocarditis, PCR remained negative in a blood culture-negative case for which no bacteria were seen at histological analysis and in one case with visualization of cocci and blood cultures positive for Enterococcus faecalis. Ten clinical diagnoses of possible infective endocarditis were ruled out by histopathological analysis of the valves and subsequently by PCR. PCR was negative in 13 of the 14 patients in whom infective endocarditis was rejected on clinical grounds; the other patient was found to have Coxiella burnetii infective endocarditis on the basis of PCR and histopathological analysis and was subsequently included in the group of 29 definite cases. In total, PCR contributed to the diagnosis and management of infective endocarditis in 6 of 29 (20%) cases. PMID- 12574280 TI - Methods for identification of Staphylococcus aureus isolates in cases of bovine mastitis. AB - A total of 272 staphylococcal isolates from cases of bovine mastitis (159 Staphylococcus aureus) belonging to 12 different species were identified with ID32 STAPH galleries, and 51 of them were confirmed by 16S rRNA gene (rrs) sequencing. The same isolates were examined for their hemolytic activity on sheep blood agar, DNase activity, and coagulase activity and with two rapid identification kits (Slidex Staph Plus kit and RAPIDEC Staph from Bio-Merieux). The results of this study confirm those obtained by other groups with hemolysis, DNase, and coagulase. Only 50% of S. aureus isolates from mastitis cases show coagulase activity after 4 h of incubation, and a 24-h incubation is necessary for the full sensitivity of this test. In contrast to results from other studies with human isolates, the Slidex Staph Plus kit was not sensitive enough for the identification of S. aureus from bovine mastitis samples. The aurease test of the RAPIDEC Staph kit showed 100% sensitivity and 100% specificity. Used in conjunction with hemolysis patterns, the RAPIDEC Staph kit is therefore very well adapted to rapid, efficient, and cost-effective identification of S. aureus in cultures from bovine mastitis samples. Sequencing of rrs genes also proved very efficient in identifying the Staphylococcus species encountered in these samples and confirming phenotypical identification results with unsatisfactory scores. With continuously improving technologies and decreasing costs, genetic identification methods like rrs gene sequencing will soon find a place in routine veterinary diagnostics. PMID- 12574281 TI - Occurrence of extended-spectrum and AmpC beta-lactamases in bloodstream isolates of Klebsiella pneumoniae: isolates harbor plasmid-mediated FOX-5 and ACT-1 AmpC beta-lactamases. AB - We tested 190 Klebsiella pneumoniae bloodstream isolates recovered from 189 patients in 30 U.S. hospitals in 23 states to determine the occurrence of extended-spectrum beta-lactamase (ESBL) and AmpC beta-lactamase producers. Based on growth inhibition by clavulanic acid by disk and MIC test methods, 18 (9.5%) of the isolates produced ESBLs. Although the disk diffusion method with standard breakpoints identified 28 cefoxitin-nonsusceptible isolates, only 5 (18%) of these were confirmed as AmpC producers. Of two AmpC confirmatory tests, the three dimensional extract test was easier to perform than was the double-disk approximation test using a novel inhibitor, Syn2190. Three of the five AmpC producers carried the bla(FOX-5) gene, while the other two isolates harbored the bla(ACT-1) gene. All AmpC genes were transferable. In vitro susceptibility testing with standard inocula showed that all five AmpC-producing strains were susceptible to cefepime, imipenem, and ertapenem but that with a high inoculum, more of these strains were susceptible to the carbapenems than to cefepime. All but 1 of 14 screen-positive AmpC nonproducers (and ESBL nonproducers) were susceptible to ceftriaxone and cefepime at the standard inoculum as were 6 of 6 isolates that were randomly selected and tested with a high inoculum. These results indicate that (i). a significant number of K. pneumoniae bloodstream isolates harbor ESBL or AmpC beta-lactamases, (ii). confirmatory tests are necessary to identify true AmpC producers, and (iii). in vitro, carbapenems are active against AmpC-producing strains of K. pneumoniae. PMID- 12574282 TI - Specimen processing and concentration of Chlamydia trachomatis added can influence false-negative rates in the LCx assay but not in the APTIMA Combo 2 assay when testing for inhibitors. AB - Inhibitors in clinical specimens can be detected by adding the target of nucleic acid amplification to the sample. Introduction of a Chlamydia trachomatis L2 434 preparation containing 12 elementary bodies (EBs) into first-void urine (FVU) from 225 nonpregnant women and 190 pregnant women before specimen processing by the assays produced false-negative rates of 0.48% (2 of 415 specimens) and 13% (44 of 338 specimens) by the APTIMA Combo 2 and the Chlamydia LCx tests, respectively. Reducing the amount of C. trachomatis added to one EB, a concentration closer to the APTIMA Combo 2 test cutoff, for a subset of 244 FVU specimens increased the number of specimens with false-negative results by the APTIMA Combo 2 assay to 7 (2.9%), suggesting that the strength of the input C. trachomatis per specimen has an influence on the number of specimens with false negative results. Repeat testing after overnight storage and dilution decreased the APTIMA Combo 2 test false-negative rates to 0% (0 of 415 specimens) with the stronger inoculum and 0.8% (2 of 244 specimens) with the weaker inoculum; the false-negative rate of the LCx assay was reduced to 5.4% (18 of 334 specimens). When an additional 70 FVU specimens from women to which 12 EBs were added before specimen processing were tested by the LCx assay, 34 specimens had false-negative results, whereas 21 specimens had false-negative results when the C. trachomatis EBs were introduced after processing. Nine of the 21 specimens to which EBs were added after processing and all of the 34 urine specimens to which the target was added before processing remained falsely negative on repeat testing at a 1:2 dilution, suggesting that input C. trachomatis DNA was lost during processing by the LCx assay. In contrast, the APTIMA Combo 2 assay appears to have a higher sensitivity and either lost little nucleic acid during processing or demonstrated few problems with inhibitors of transcription-mediated amplification. PMID- 12574283 TI - Apophysomyces elegans: an emerging zygomycete in India. AB - Apophysomyces elegans was considered a rare but medically important zygomycete. We analyzed the clinical records of eight patients from a single center in whom zygomycosis due to A. elegans was diagnosed over a span of 25 months. We also attempted a DNA-based method for rapid identification of the fungi and looked for interstrain polymorphism using microsattelite primers. Three patients had cutaneous and subcutaneous infections, three had isolated renal involvement, one had rhino-orbital tissue infection, and the final patient had a disseminated infection involving the spleen and kidney. Underlying illnesses were found in two patients, one with diabetes mellitus and the other with chronic alcoholism. A history of traumatic implantation was available for three patients. All except two of the patients responded to surgical and/or medical therapy; the diagnosis for the two exceptions was made at the terminal stage of infection. Restriction enzyme (MboI, MspI, HinfI) digestion of the PCR-amplified internal transcribed spacer region helped with the rapid and specific identification of A. elegans. The strains could be divided into two groups according to their patterns, with clustering into one pattern obtained by using microsatellite [(GTG)(5) and (GAC)(5)] PCR fingerprinting. The study highlights the epidemiology, clinical spectrum, and diagnosis of emerging A. elegans infections. PMID- 12574284 TI - Multicenter quality control for the detection of hepatitis C virus RNA in seminal plasma specimens. AB - The discrepant results available in the literature about the presence of hepatitis C virus (HCV) RNA in seminal plasma of men chronically infected by this agent are related, at least in part, to the molecular techniques used and particularly to the wide range of protocols dedicated to RNA extraction. In order to evaluate these protocols and to standardize the method of detection of HCV RNA in this fluid, a panel of coded specimens was tested blindly in 12 French laboratories; it included 14 seminal plasma specimens and four water controls spiked with HCV RNA ranging from 10 to 20000 IU/ml and two HCV-negative seminal plasma specimens. The extraction step was performed according to methods using either silica beads (NucliSens [Organon Teknika S.A., Fresnes, France]; RNA viral kit [Qiagen, Courtaboeuf, France]) or guanidinium thiocyanate (Amplicor HCV assay; Roche Diagnostics, Meylan, France), preceded or not by a centrifugation of the seminal plasma. For the amplification step, all the laboratories performed the same reverse transcription-PCR technique (Amplicor HCV Cobas assay). The percentage of correct results ranged from 53.3 to 100, the poorest results being obtained when no centrifugation step preceded the Amplicor extraction protocol. The rate of correct results was significantly higher in laboratories using a preliminary centrifugation of the specimen (P = 0.034 by chi-square test). By contrast, the overall number of correct results was not correlated to the initial volume of sample used for the test. These results allowed us to validate standardized techniques adapted to the performance of this test on a routine basis, especially in men infected with HCV and involved in programs of medically assisted reproduction. PMID- 12574285 TI - Analysis of an IS2404-based nested PCR for diagnosis of Buruli ulcer disease in regions of Ghana where the disease is endemic. AB - Mycobacterium ulcerans causes Buruli ulcer disease (BUD), an ulcerative skin disease emerging mainly in West Africa. Laboratory confirmation of BUD is complicated as no "gold standard" for diagnosis exists. A nested primer PCR based on IS2404 has shown promise as a diagnostic assay. We evaluated the IS2404-based PCR to detect M. ulcerans DNA in tissue specimens from 143 BUD patients diagnosed according to the World Health Organization BUD clinical case definition in Ghana. Comparisons were made with culture and histopathology results. Variables influencing detection rate tested in this PCR protocol included the amount of tissue used and the stage of disease. The nested PCR was repeated on DNA extracted from a different part of the same biopsy specimen of 21 culture positive samples. Of all 143 specimens, 107 (74.8%; 95% confidence interval, 68 to 82%) showed the presence of M. ulcerans DNA by PCR. Of the 78 histology confirmed BUD patient samples, 64 (83%) were PCR positive. Detection rates were influenced neither by the amount of tissue processed for PCR nor by the stage of disease (preulcerative or ulcerative). Taken together, the two nested PCR tests on the subset of 21 culture-positive samples were able to detect M. ulcerans DNA in all 21 culture-confirmed patients. For future studies, small tissue samples, e.g., punch biopsy samples, might be sufficient for case confirmation. PMID- 12574286 TI - Use of multienzyme multiplex PCR amplified fragment length polymorphism typing in analysis of outbreaks of multiresistant Klebsiella pneumoniae in an intensive care unit. AB - We developed and optimized a new modified amplified fragment length polymorphism (AFLP) typing method to obtain a multibanding fingerprint that can be separated by agarose gel electrophoresis. Both to maximize the discriminatory power and to facilitate the computer-assisted analysis, bacterial DNA was digested with four different restriction enzymes. After ligation of adaptors to the DNA fragments, PCR testing of various single primers was performed. Two single primers that gave optimal results with regard to band resolution and discriminatory power were selected and combined. The computer-assisted analysis of fingerprint patterns was performed with Pearson's product-moment correlation values of densitometric curves, without assigning bands to peaks. Thus, the analysis is not subject to human interpretation errors. With this method, we investigated two outbreaks of multiresistant Klebsiella pneumoniae in an intensive care unit and various sporadic isolates of K. pneumoniae and Klebsiella oxytoca. Cluster analysis of isolates analyzed in different experiments and on different gels showed that fingerprint patterns clustered correctly according to subspecies or to the outbreaks. Multienzyme multiplex PCR AFLP revealed that the first outbreak was caused by two different types of strains. Outbreak two was caused by yet another strain of K. pneumoniae. In conclusion, the typing method used here is easy to perform and highly reproducible, and due to generation of complex banding patterns, it has a higher discriminatory power. Furthermore, the multienzyme multiplex PCR fingerprints are easy to analyze, and a reliable database can be stored in the computer to facilitate comparison of future isolates of Klebsiella spp. The method can be performed in every clinical microbiology laboratory. PMID- 12574287 TI - Evaluation of four commercially available rapid serologic tests for diagnosis of leptospirosis. AB - Four rapid tests for the serologic diagnosis of leptospirosis were evaluated, and the performance of each was compared with that of the current standard, the microscopic agglutination test (MAT). The four rapid tests were a microplate immunoglobulin M (IgM)-enzyme-linked immunosorbent assay (ELISA), an indirect hemagglutination assay (IHA), an IgM dipstick assay (LDS), and an IgM dot-ELISA dipstick test (DST). A panel of 276 sera from 133 cases of leptospirosis from four different geographic locations was tested as well as 642 sera from normal individuals or individuals with other infectious or autoimmune diseases. Acute phase sera from cases (n = 148) were collected or=15 days after onset (median = 29.1). By a traditional method (two-by-two contingency table), the sensitivities for detection of leptospirosis cases were 93.2% by LDS, 92.5% by DST, 86.5% by ELISA, and 79.0% by IHA. Specificity was 98.8% by DST, 97% by ELISA and MAT, 95.8% by IHA, and 89.6% by LDS. With a latent class analysis (LCA) model that included all the rapid tests and the clinical case definition, sensitivity was 95.5% by DST, 94.5% by LDS, 89.9% by ELISA, and 81.1% by IHA. The sensitivity and specificity estimated by the traditional methods were quite close to the LCA estimates. However, LCA allowed estimation of the sensitivity of the MAT (98.2%), which traditional methods do not allow. For acute-phase sera, sensitivity was 52.7% by LDS, 50.0% by DST, 48.7% by MAT and ELISA, and 38.5% by IHA. The sensitivity for convalescent-phase sera was 93.8% by MAT, 84.4% by DST, 83.6% by LDS, 75.0% by ELISA, and 67.2% by IHA. A good overall correlation with the MAT was obtained for each of the assays, with the highest concordance being with the DST (kappa value, 0.85; 95% confidence interval [CI], 0.8 to 0.90). The best correlation was between ELISA and DST (kappa value, 0.86; 95% CI, 0.81 to 0.91). False-positive LDS results were frequent (>or=20%) in sera from individuals with Epstein-Barr virus, human immunodeficiency virus, and periodontal disease and from healthy volunteers. The ease of use and significantly high sensitivity and specificity of DST and ELISA make these good choices for diagnostic testing. PMID- 12574288 TI - Rapid detection of adenovirus in throat swab specimens by PCR during respiratory disease outbreaks among military recruits. AB - We evaluated the performance of a generic PCR test to detect adenoviruses (AdV) in throat swab specimens collected from asymptomatic and ill military recruits with acute respiratory disease. Samples (n = 210) were collected at entry to basic training and at the time of large outbreaks of AdV-associated acute respiratory disease among military recruits at Fort Jackson, South Carolina, from 1997 to 1998. Compared to cell culture, a sensitivity of 99% and a specificity of 98% were noted for the PCR method to detect AdV in throat swabs. Similar results were obtained with or without DNA extraction, suggesting the absence of significant inhibitors for the PCR method in throat swab samples. No AdV was detected by culture or PCR in throat swabs from healthy recruits, suggesting the absence of latency or asymptomatic shedding. Throat swab specimens proved to be adequate, noninvasive samples to rapidly diagnose respiratory disease in young adults. This generic direct PCR proved to be a useful test for the rapid diagnosis of AdV-associated respiratory disease, detecting all serotypes tested to date and furnishing results within 6 h of specimen arrival. The use of this direct, rapid, sensitive, and specific assay would assist health care providers and public health practitioners in the early diagnosis, management, and control of AdV-associated respiratory disease. PMID- 12574289 TI - Detection of Helicobacter pylori in paraffin-embedded and in shock-frozen gastric biopsy samples by fluorescent in situ hybridization. AB - We report on the successful application of fluorescent in situ hybridization for detection of Helicobacter pylori and determination of its clarithromycin susceptibility in formalin-fixed and paraffin-embedded gastric biopsy specimens that had been prepared for pathological examination. This method is useful when results from conventional culturing with antibiotic susceptibility testing are not available. PMID- 12574290 TI - Rapid detection of rifampin resistance in Mycobacterium tuberculosis isolates by heteroduplex analysis and determination of rifamycin cross-resistance in rifampin resistant isolates. AB - Direct heteroduplex analysis and a universal heteroduplex generator assay were performed to detect rifampin resistance rapidly in Turkish Mycobacterium tuberculosis isolates. Cross-resistance to rifapentine, rifabutin, and rifalazil was investigated. A relationship between specific mutations and resistance patterns, which can guide the choice of an appropriate therapeutic regimen for tuberculosis patients, was identified. PMID- 12574291 TI - Optimum detection times for bacteria and yeast species with the BACTEC 9120 aerobic blood culture system: evaluation for a 5-year period in a Turkish university hospital. AB - We tracked and documented the time of positivity of blood cultures by using the BACTEC 9120 (Becton Dickinson Diagnostic Instrument Systems) blood culture system over a 5-year study period. A 7-day protocol of the incubation period was selected, and a total of 11156 blood cultures were evaluated. The clinically significant microorganisms (32.95%) were isolated in 3676 specimens. Gram positive and -negative bacterial isolation rates were found to be 41.07 and 44.88%, respectively. Yeasts were found in 14.03% of all pathogens. Both the false-positivity and -negativity rates were very low (0.1 and 0.3%, respectively). The mean detection times for all of the pathogens were determined to be 19.45 h. Yeasts, nonfermentative gram-negative bacteria, and Brucella melitensis strains were isolated within 5 days. By taking these data into account, we decided to establish a 5-day-incubation protocol in our laboratory instead of the 7 days that are commonly used. PMID- 12574292 TI - Spread of integron-associated VIM-type metallo-beta-lactamase genes among imipenem-nonsusceptible Pseudomonas aeruginosa strains in Greek hospitals. AB - Fifty-eight imipenem-nonsusceptible (MIC >or= 8 microg/ml) Pseudomonas aeruginosa strains isolated during May 2001 in 15 Greek hospitals were studied. Thirty-six isolates derived from nine hospitals carried VIM-type metallo-beta-lactamase genes, as found by PCR. In 34 isolates, bla(VIM) was associated with class 1 integrons of various sizes. DNA sequencing indicated the presence of bla(VIM-2) gene cassettes in a variety of integron structures. Random amplified polymorphic DNA typing suggested diversity of the bla(VIM)-positive strains. Synergy between 2-mercaptoacetic acid and imipenem indicated carbapenemase activity in 26 bla(VIM)-positive strains. PMID- 12574293 TI - Identification of dermatophyte species by 28S ribosomal DNA sequencing with a commercial kit. AB - We have shown that dermatophyte species can be easily identified on the basis of a DNA sequence encoding a part of the large-subunit (LSU) rRNA (28S rRNA) by using the MicroSeq D2 LSU rRNA Fungal Sequencing Kit. Two taxa causing distinct dermatophytoses were clearly distinguished among isolates of the Trichophyton mentagrophytes species complex. PMID- 12574294 TI - Detection of Candida albicans mRNA from formalin-fixed, paraffin-embedded mouse tissues by nested reverse transcription-PCR. AB - Histopathology archives represent a vast source of infectious disease specimens that can be used to elucidate important disease processes. In this report, we describe a method to detect Candida albicans gene expression from infected, formalin-fixed, paraffin-embedded mouse tissue. By use of glass beads to break open the fungal cells and proteinase K treatment, RNA was extracted routinely from tissue sections that had been fixed for up to 72 h. Upon reverse transcription of the RNA and nested PCR, the procedure detected C. albicans "housekeeping" and putative virulence genes. PMID- 12574295 TI - Evaluation of the specificities of five DNA amplification methods for the detection of Neisseria gonorrhoeae. AB - The intragenus specificities of five molecular diagnostic methods for Neisseria gonorrhoeae were determined. Three assays were considered suboptimal. Molecular detection of N. gonorrhoeae from sites where other Neisseria spp. commonly occur or from any site in low-prevalence settings should be confirmed by a test targeting a different genetic locus. PMID- 12574296 TI - Clinical utility of urinary antigen detection for diagnosis of community acquired, travel-associated, and nosocomial legionnaires' disease. AB - The clinical utility of Legionella urinary antigen assays for the diagnosis of Legionnaires' disease was assessed by using samples from 317 culture-proven cases. The sensitivities of the Binax enzyme immunoassay (EIA) and Biotest EIA were found to be 93.7 and 94.4% for travel-associated infection and 86.5 and 76.0% for community-acquired infection but only 44.2 and 45.7% for nosocomially acquired infection, respectively. PMID- 12574297 TI - Diagnosis of enterovirus infection by genus-specific PCR and enzyme-linked immunosorbent assays. AB - PCR for the diagnosis of enterovirus infections is resource intensive but is increasingly used due to wide availability. Enzyme-linked immunosorbent assays (ELISAs) that detect heterotypical antibodies against enterovirus immunoglobulin M (IgM), IgA, and IgG were compared with reverse transcription-PCR by using primers specific to the 5' untranslated regions of 60 enterovirus species. The ELISAs were less sensitive than the PCR, and only the ELISA for IgM was highly specific. When retrospective diagnosis is important or when specimens are unsuitable for PCR, the ELISA has a limited role if PCR is not available. PMID- 12574298 TI - AFLMP1 encodes an antigenic cel wall protein in Aspergillus flavus. AB - We have previously reported the cloning and characterization of the MP1 gene in Penicillium marneffei and the AFMP1 gene in Aspergillus fumigatus and their use for serodiagnosis of penicilliosis and aspergilloma and invasive aspergillosis, respectively. In this study, we describe the cloning of the AFLMP1 gene, which encodes the homologous antigenic cell wall protein in Aspergillus flavus, the most common Aspergillus species associated with human disease in our locality and in other Asian countries and the second most common Aspergillus species associated with human disease in Western countries. AFLMP1 codes for a protein, Aflmp1p, of 273 amino acid residues, with a few sequence features that are present in Mp1p and Afmp1p, the homologous antigenic cell wall proteins in P. marneffei and A. fumigatus, respectively, as well as several other cell wall proteins of Saccharomyces cerevisiae and Candida albicans. It contains a serine- and threonine-rich region for O glycosylation, a signal peptide, and a putative glycosylphosphatidylinositol attachment signal sequence. Specific anti-Aflmp1p antibody was generated with recombinant Aflmp1p protein purified from Escherichia coli to allow further characterization of Aflmp1p. Indirect immunofluorescence analysis indicated that Aflmp1p is present on the surface of the hyphae of A. flavus. Finally, it was observed that patients with aspergilloma and invasive aspergillosis due to A. flavus develop a specific antibody response against Aflmp1p. This suggested that the recombinant protein and its antibody may be useful for serodiagnosis in patients with aspergilloma or invasive aspergillosis, and the protein may represent a good cell surface target for host humoral immunity. PMID- 12574299 TI - Phylogeny of the genus Nocardia based on reassessed 16S rRNA gene sequences reveals underspeciation and division of strains classified as Nocardia asteroides into three established species and two unnamed taxons. AB - Conventional identification of Nocardia in the routine laboratory remains problematic due to a paucity of reliable phenotypic tests and due to the yet unresolved taxonomy of strains classified as belonging to the species Nocardia asteroides, which comprises the type strain and isolates with drug pattern types II and VI. The 16S rRNA gene of 74 representative strains of the genus Nocardia, encompassing 25 established species, was sequenced in order to provide a molecular basis for accurate species identification and with the aim of reassessing the phylogeny of taxons assigned to the species N. asteroides. The result of this phylogenetic analysis confirms that the interspecies heterogeneity of closely related nocardial species can be considerably low (a sequence divergence of only 0.5% was found between N. paucivorans and N. brevicatena). We observed a sequence microheterogeneity (sequence divergence of fewer than five bases) in 8 of 11 species of which more than one strain in the species was studied. At least 10 taxons were found that merit description as new species. Strains previously classified as N. asteroides fell into five distinct phylogenetic groups: the type strain cluster (N. asteroides sensu strictu), N. abscessus, N. cyriacigeorgica, and two clusters closely related to N. carnea or N. flavorosea. The strains within the latter two groups probably represent new species, pending further genetic and phenotypic evaluation. Restricted phenotypic data revealed that N. abscessus, N. cyriacigeorgica, and the two Nocardia species taxons are equivalent to drug patterns I, VI, and II, respectively. In the future, these data will help in finding species-specific markers after adoption of a more precise nomenclature for isolates closely related to N. asteroides and unravel confusing phenotypic data obtained in the past for unresolved groups of strains that definitely belong to separate taxons from a phylogenetic point of view. PMID- 12574300 TI - Catheter-related sepsis due to Rhodotorula glutinis. AB - We describe a central venous catheter-related (Port-A-Cath; Smiths Industries Medical Systems [SIMS] Deltec, Inc., St. Paul, Minn.) infection caused by Rhodotorula glutinis in a 51-year-old man with nasopharyngeal carcinoma. He was treated with fluconazole for 8 weeks and had the catheter removed. Two isolates of R. glutinis recovered from blood specimens (one obtained via peripheral veins and one via the catheter) before administration of fluconazole and one recovered from the removed catheter 17 days after initiation of fluconazole therapy exhibited high-level resistance to fluconazole (MICs, >256 microg/ml). These three isolates were found to belong to a single clone on the basis of identical antibiotypes determined by the E test (PDM Epsilometer; AB Biodisk, Solna, Sweden) and biotypes determined by API ID32 C (bioMerieux, Marcy I'Etoile, France) and their identical random amplified polymorphic DNA patterns. PMID- 12574301 TI - Genetic analysis of Crimean-Congo hemorrhagic fever virus in Russia. AB - Genetic analysis of wild-type Crimean-Congo hemorrhagic fever (CCHF) virus strains recovered in the European part of Russia was performed. Reverse transcriptase PCR followed by direct sequencing was used to recover partial sequences of the CCHF virus medium (M) genome segment (M segment) from four pools of Hyalomma marginatum ticks and six human patients. Phylogenetic analysis of the M-segment sequences from Russian strains revealed a close relatedness of the strains (nucleotide sequence diversity, 90% IF, and this predominance of cells with apparently a single IF rearrangement continued in all organs until approximately DG45, which corresponds to the time when lymphopoiesis begins in the bone marrow. Thereafter, the proportion of IF rearrangements drops to approximately 71%, i.e., the value predicted whether VDJ rearrangement is random and both chromosomes were involved. Unlike other tissues, VDJs recovered from thymus after DG50 display a pattern suggesting no selection for IF rearrangements. Regardless of differences in the proportion of IF rearrangements, we observed no significant age- or tissue-dependent changes in CDR3 diversity, N region additions, or other characteristics of fetal VDJs during ontogeny. These findings indicate there are multiple sites of B cell lymphogenesis in fetal piglets and differences in the frequency of productive VDJ rearrangements at various sites. We propose the latter to result from differential selection or a developmentally dependent change in the intrinsic mechanism of VDJ rearrangement. PMID- 12574343 TI - The Th2 cell cytokines IL-4 and IL-13 regulate found in inflammatory zone 1/resistin-like molecule alpha gene expression by a STAT6 and CCAAT/enhancer binding protein-dependent mechanism. AB - The onset of allergic inflammation in the lung is driven by a complex genetic program. This study shows that found in inflammatory zone (FIZZ)1 and FIZZ2, but not FIZZ3, gene expression was up-regulated 6 h after Ag challenge in a mouse model of acute pulmonary inflammation. Induction of both genes was abolished in allergen-challenged STAT6-deficient mice. FIZZ1, but not FIZZ2, mRNA was up regulated upon incubation of the myeloid cell line BMnot with IL-4. The promoter region of FIZZ1 contains functional binding sites for STAT6 and C/EBP. FIZZ1 promoter reporter gene constructs responded to IL-4 and IL-13 stimulation in transiently transfected cells. Point mutations in the STAT6 or the C/EBP site led to loss of cytokine responsiveness indicating that IL-4-mediated induction of murine FIZZ1 is orchestrated by the coordinate action of STAT6 and C/EBP. It is concluded that the expression of the genes encoding FIZZ1 and FIZZ2, but not FIZZ3, is induced in allergen-challenged lungs in a STAT6-dependent fashion. STAT6 directly regulates IL-4- and IL-13-triggered induction of FIZZ1 expression at the transcriptional level by cooperation with C/EBP. Induction of FIZZ2 gene expression most likely occurs independent of a direct effect by these cytokines and may be due to indirect STAT6-driven mechanisms. PMID- 12574344 TI - Cooperation of TNF family members CD40 ligand, receptor activator of NF-kappa B ligand, and TNF-alpha in the activation of dendritic cells and the expansion of viral specific CD8+ T cell memory responses in HIV-1-infected and HIV-1 uninfected individuals. AB - Members of the TNF superfamily have been shown to be instrumental in enhancing cell-mediated immune responses, primarily through their interactions with dendritic cells (DCs). We systematically evaluated the ability of three TNF superfamily molecules, CD40 ligand (CD40L), receptor activator of NF-kappaB ligand (RANKL), and TNF-alpha, to expand ex vivo EBV-specific CTL responses in healthy human individuals and ex vivo HIV-1-specific CTL responses in HIV-1 infected individuals. In both groups of individuals, we found that all three TNF family molecules could expand CTL responses, albeit at differing degrees. CD40L treatment alone was better than RANKL or TNF-alpha alone to mature DCs and to expand CTL. In healthy volunteers, TNF-alpha or RANKL could cooperate with CD40L to maximize the ability of DCs to expand virus-specific CTL responses. In HIV-1 infection, cooperative effects between TNF-alpha or RANKL in combination with CD40L were variable. TNF-alpha and RANKL cooperated with CD40L via differing mechanisms, i.e., TNF-alpha enhanced IL-12 production, whereas RANKL enhanced survival of CD40L-stimulated DCs. These findings demonstrate that optimal maturation of DCs requires multiple signals by TNF superfamily members that include CD40L. In HIV-1 infection, DCs may only require CD40L to maximally expand CTL. Finally, CTL responses were higher in CD4(+) T cell-containing conditions even in the presence of TNF family molecules, suggesting that CD4(+) T cells can provide help to CD8(+) T cells independently of CD40L, RANKL, or TNF-alpha. PMID- 12574345 TI - Suppressive DNA vaccination in myelin oligodendrocyte glycoprotein peptide induced experimental autoimmune encephalomyelitis involves a T1-biased immune response. AB - Vaccination with DNA encoding a myelin basic protein peptide suppresses Lewis rat experimental autoimmune encephalomyelitis (EAE) induced with the same peptide. Additional myelin proteins, such as myelin oligodendrocyte glycoprotein (MOG), may be important in multiple sclerosis. Here we demonstrate that DNA vaccination also suppresses MOG peptide-induced EAE. MOG(91-108) is encephalitogenic in DA rats and MHC-congenic LEW.1AV1 (RT1(av1)) and LEW.1N (RT1(n)) rats. We examined the effects of DNA vaccines encoding MOG(91-108) in tandem, with or without targeting of the hybrid gene product to IgG. In all investigated rat strains DNA vaccination suppressed clinical signs of EAE. There was no requirement for targeting the gene product to IgG, but T1-promoting CpG DNA motifs in the plasmid backbone of the construct were necessary for efficient DNA vaccination, similar to the case in DNA vaccination in myelin basic protein-induced EAE. We failed to detect any effects on ex vivo MOG-peptide-induced IFN-gamma, TNF-alpha, IL-6, IL 4, IL-10, and brain-derived neurotropic factor expression in splenocytes or CNS derived lymphocytes. In CNS-derived lymphocytes, Fas ligand expression was down regulated in DNA-vaccinated rats compared with controls. However, MOG-specific IgG2b responses were enhanced after DNA vaccination. The enhanced IgG2b responses together with the requirement for CpG DNA motifs in the vaccine suggest a protective mechanism involving induction of a T1-biased immune response. PMID- 12574346 TI - TNF-related apoptosis-inducing ligand mediates tumoricidal activity of human monocytes stimulated by Newcastle disease virus. AB - The Newcastle disease virus (NDV) has antineoplastic and immunostimulatory properties, and it is currently clinically tested in anticancer therapy. However, the tumoricidal mechanisms of NDV tumor therapy are not fully understood. The results presented here demonstrate that NDV-stimulated human monocytes (Mphi) kill various human tumor cell lines and that this tumoricidal activity is mediated by TRAIL. In contrast to soluble TRAIL-R2-Fc, soluble CD95-Fc and TNF-R2 Fc showed only minimal blocking of the antitumor effect. TRAIL expression is induced on human Mphi after stimulation with NDV and UV-inactivated NDV. These results show that TRAIL induction on human Mphi after NDV stimulation is independent from viral replication and that TRAIL mediates the tumoricidal activity of NDV-stimulated human Mphi. PMID- 12574347 TI - Antigen density presented by dendritic cells in vivo differentially affects the number and avidity of primary, memory, and recall CD8+ T cells. AB - We studied the size and avidity of primary and recall CD8(+) T cell responses in vivo in mice immunized with dendritic cells presenting different densities of a MHC class I-restricted peptide. Increasing the epitope density on a fixed number of dendritic cells increased the size of the primary response, yet had no influence on the avidity of the effector cells. However, epitope density-based selection of cells with different avidities was evident in the subsequent memory population, and in recall responses. Additionally, mice primed with different peptide densities had similarly sized quiescent memory and recall responses. Our findings provide evidence for an important role for epitope density in the selection of T cells in vivo. PMID- 12574348 TI - Naturally occurring human IgM antibody that binds B7-DC and potentiates T cell stimulation by dendritic cells. AB - A human IgM Ab, serum-derived human IgM 12 (sHIgM12), is identified that binds mouse and human dendritic cells (DC), inducing dramatic immunopotentiation following treatment of the mouse DC in vitro. Competition, transfection, and knockout studies identified the ligand on mouse DC as the costimulatory molecule family member B7-DC. Potent T cell responses are stimulated by Ag-pulsed DC treated with the sHIgM12 Ab in vitro and upon adoptive transfer of Ab-treated Ag pulsed DC into animals. The multivalent structure of pentameric IgM provides the potential for cross-linking cell surface targets, endowing the soluble Abs with biological potential not normally associated with immune function. The ability of the sHIgM12 Ab to potentiate the immune response is dependent on the multimeric structure of IgM, as bivalent monomers do not retain this property. Furthermore, pretreatment of DC with IgM monomers blocks subsequent potentiation by intact IgM pentamers, an indication that cross-linking of B7-DC on the cell surface is critical for potentiation of Ag presentation. These findings imply that, in addition to known costimulatory roles, B7-DC can function as a receptor for signals delivered by cells expressing B7-DC ligands. PMID- 12574349 TI - Temperature effect on IgE binding to CD23 versus Fc epsilon RI. AB - A chimeric soluble CD23, consisting of the extracellular domain of mouse CD23 and a modified leucine zipper (lz-CD23), has been shown to inhibit IgE binding to the FcepsilonRI. A similar human CD23 construct was also shown to inhibit binding of human IgE to human FcepsilonRI. In both systems, the inhibition was found to be temperature dependent; a 10-fold molar excess of lz-CD23 gave 90-98% inhibition at 4 degrees C, dropping to 20-30% inhibition at 37 degrees C. Surface plasmon resonance analysis of lz-CD23 binding to an IgE-coated sensor chip suggested that the effective concentration of lz-CD23 was lower at the higher temperatures. Analysis of (125)I-IgE binding to CD23(+)-Chinese hamster ovary cells also indicated that increased temperature resulted in a lower percentage of IgE capable of interacting with CD23. In contrast, IgE interacts more effectively with FcepsilonRI(+)-rat basophilic leukemia cells at 37 degrees C compared with 4 degrees C. The results support the concept that the open and closed IgE structures found by crystallography interact differently with the two IgE receptors and suggest that temperature influences the relative percentage of IgE in the respective structural forms. Changes in CD23 oligomerization also plays a role in the decreased binding seen at physiological temperatures. PMID- 12574350 TI - Role of double-negative regulatory T cells in long-term cardiac xenograft survival. AB - A novel subset of CD3(+)CD4(-)CD8(-) (double negative; DN) regulatory T cells has recently been shown to induce donor-specific skin allograft acceptance following donor lymphocyte infusion (DLI). In this study, we investigated the effect of DLI on rat to mouse cardiac xenotransplant survival and the ability of DN T cells to regulate xenoreactive T cells. B6 mice were given either DLI from Lewis rats, a short course of depleting anti-CD4 mAb, both DLI and anti-CD4 treatment together, or left untreated. DLI alone did not prolong graft survival when compared with untreated controls. Although anti-CD4-depleting mAb alone significantly prolonged graft survival, grafts were eventually rejected by all recipients. However, the combination of DLI and anti-CD4 treatment induced permanent cardiac xenograft survival. We demonstrate that recipients given both DLI and anti-CD4 treatment had a significant increase in the total number of DN T cells in their spleens when compared with all other treatment groups. Furthermore, DN T cells harvested from the spleens of DLI plus anti-CD4-treated mice could dose-dependently inhibit the proliferation of syngeneic antidonor T cells. Suppression mediated by these DN T cells was specific for antidonor T cells as T cells stimulated by third party Ags were not suppressed. These results demonstrate for the first time that a combination of pretransplant DLI and anti-CD4-depleting mAb can induce permanent survival of rat to mouse cardiac xenografts and that DN T regulatory cells play an important role in preventing long-term concordant xenograft rejection through the specific suppression of antidonor T cells. PMID- 12574351 TI - Immunoglobulin superantigen protein L induces IL-4 and IL-13 secretion from human Fc epsilon RI+ cells through interaction with the kappa light chains of IgE. AB - Peptostreptococcus magnus protein L is a multidomain bacterial surface protein that correlates with virulence. It consists of up to five homologous Ig-binding domains (B1-B5) that interact with the variable domain of Ig kappa L chains. Intact protein L stimulates the synthesis and the release of IL-4 and IL-13 from human basophils in vitro. A protein L fragment covering the Ig-binding domains B1 B4 also induced IL-4 and IL-13 release from basophils. There was an excellent correlation (r(s) = 0.82; p < 0.001) between the maximal percent IL-4 release induced by protein L and that induced by anti-IgE and between intact protein L and the B1-B4 fragment (r(s) = 0.90; p < 0.01). Removal of IgE bound to basophils markedly reduced the IL-4 release induced by anti-IgE, protein L, and B1-B4. Preincubation of basophils with protein L or anti-IgE caused complete cross desensitization to subsequent challenge with the heterologous stimulus. IgE purified from myeloma patients PS and PP (lambda chains) blocked anti-IgE-induced IL-4 release, but not the releasing activity of protein L. In contrast, IgE purified from myeloma patient ADZ (kappa chains) blocked both anti-IgE- and protein L-induced secretion. Cyclosporin A, but not cyclosporin H, inhibited protein L-induced release of IL-4 and IL-13 from basophils. Thus, protein L acts as a bacterial Ig superantigen to induce the synthesis and release of IL-4 and IL 13 from basophils by interacting with kappa L chains of the IgE isotype. PMID- 12574352 TI - H2-M3-restricted memory T cells: persistence and activation without expansion. AB - H2-M3-restricted T cells respond more rapidly to primary Listeria monocytogenes infection than conventional MHC class Ia-restricted T cells. Reinfection with L. monocytogenes, while inducing explosive proliferation of H2-K(d)-restricted T cells, does not stimulate significant expansion of H2-M3-restricted CTL. These disparate responses to reinfection are apparent within 5 days of primary L. monocytogenes infection. However, H2-M3-restricted memory T cells are generated, and are indistinguishable from classically restricted T cells in terms of cell surface memory markers and longevity. Early responses of H2-M3- and H2-K(d) restricted memory T cells to reinfection are similar, with increases in size and expression of activation markers. Interestingly, priming of H2-M3-restricted T cells with an L. monocytogenes-derived N-formyl peptide plus anti-CD40 generates memory T cells that expand upon re-exposure to Ag during L. monocytogenes infection. Our data indicate that disparate H2-M3- and MHC class Ia-restricted memory T cell responses reflect intrinsic differences between these T cell populations. Although distinct proliferative programs appear to be hardwired in these populations during primary L. monocytogenes infection, under different inflammatory circumstances M3-restricted T cell populations can maintain the ability to expand upon re-exposure to Ag. PMID- 12574353 TI - Stat signals release activated naive Th cells from an anergic checkpoint. AB - Activation of naive Th lymphocytes by the TCR and the costimulatory molecule, CD28, is believed to provide competent signals for differentiation to effector cells. Such activated cells proliferated and expressed IL-2, but arrested in an immature state maintained by CTLA-4. Although unresponsive to restimulation by TCR/CD28 alone, restimulation with TCR/CD28 and either Stat4- or Stat6-mediated cytokine signals rescued cells to proliferate and differentiate to the appropriately matched canonical Th subsets. Addition of IL-4 at defined periods revealed that naive T cells were receptive to IL-4-mediated differentiation for up to 3 days after their initial priming. A Stat-dependent anergic checkpoint between clonal expansion and effector cell differentiation may defer the cytokine profile to be instructed at the site of infection, thus preventing the unregulated development of potentially damaging effector cells. PMID- 12574354 TI - Characterization of thymic progenitors in adult mouse bone marrow. AB - Thymic cellularity is maintained throughout life by progenitor cells originating in the bone marrow. In this study, we describe adult mouse bone cells that exhibit several features characteristic of prothymocytes. These include 1) rapid thymic engraftment kinetics following i.v. transplantation, 2) dramatic expansion of thymic progeny, and 3) limited production of hemopoietic progeny other than thymocytes. The adult mouse bone marrow population that is depleted of cells expressing any of a panel of lineage-specific Ags, stem cell Ag-1 positive, and not expressing the Thy1.1 Ag (Thy1.1(-)) (Thy1.1(-) progenitors) can repopulate the thymus 9 days more rapidly than can hemopoietic stem cells, a rate of thymic repopulation approaching that observed with transplanted thymocytes. Additionally, Thy1.1(-) progenitors expand prolifically to generate thymocyte progeny comparable in absolute numbers to those observed from parallel hemopoietic stem cell transplants, and provide a source of progenitors that spans multiple waves of thymic seeding. Nevertheless, the Thy1.1(-) population yields relatively few B cells and rare myeloid progeny posttransplant. These observations describe the phenotype of an adult mouse bone marrow population highly enriched for rapidly engrafting, long-term thymocyte progenitors. Furthermore, they note disparity in B and T cell expansion from this lymphoid progenitor population and suggest that it contains the progenitor primarily responsible for seeding the thymus throughout life. PMID- 12574355 TI - Human activation-induced cytidine deaminase is induced by IL-4 and negatively regulated by CD45: implication of CD45 as a Janus kinase phosphatase in antibody diversification. AB - Activation-induced cytidine deaminase (AID) plays critical roles in Ig class switch recombination and V(H) gene somatic hypermutation. We investigated the role of IL-4 in AID mRNA induction, the signaling transduction involved in IL-4 mediated AID induction, and the effect of CD45 on IL-4-dependent AID expression in human B cells. IL-4 was able to induce AID expression in human primary B cells and B cell lines, and IL-4-induced AID expression was further enhanced by CD40 signaling. IL-4-dependent AID induction was inhibited by a dominant-negative STAT6, indicating that IL-4 induced AID expression via the Janus kinase (JAK)/STAT6 signaling pathway. Moreover, triggering of CD45 with anti-CD45 Abs can inhibit IL-4-induced AID expression, and this CD45-mediated AID inhibition correlated with the ability of anti-CD45 to suppress IL-4-activated JAK1, JAK3, and STAT6 phosphorylations. Thus, in humans, IL-4 alone is sufficient to drive AID expression, and CD40 signaling is required for optimal AID production; IL-4 induced AID expression is mediated via the JAK/STAT signaling pathway, and can be negatively regulated by the JAK phosphatase activity of CD45. This study indicates that the JAK phosphatase activity of CD45 can be induced by anti-CD45 Ab treatment, and this principle may find clinical application in modulation of JAK activation in immune-mediated diseases. PMID- 12574356 TI - Locus-specific constitutive and cytokine-induced HLA class I gene expression. AB - Cytokine induction of the MHC class I genes increases the nascent molecules available for binding potentially antigenic peptides. The human H chain loci, HLA A, -B, and -C, encode highly homologous and polymorphic mRNAs. Here, these transcripts were resolved and measured by competitive PCR of cDNA using locus specific primers. Endothelial cells expressed many HLA-A and -B, but fewer HLA-C, transcripts. In contrast, HeLa cells expressed many HLA-A and -C, but fewer HLA B, transcripts. The inflammatory cytokines TNF-alpha, IFN-beta, and IFN-gamma induced HLA-B strongly, but HLA-A and -C weakly in both cell types. Combined treatment with IFNs and TNF further increased HLA-A and -B, but not HLA-C transcripts. The constitutive and inducible activities of transfected promoters correlated well with mRNA levels. The weak IFN response of the HLA-A2 promoter was not due to variations in the IFN consensus sequence, the site alpha, or a 3 bp insertion between them. The HLA-Cw6 promoter was less TNF responsive due to a variant kappaB enhancer, which also reduced the IFN responses. The NF-kappaB subunit RelA strongly activated the HLA-A2 and -B7 promoters but only weakly activated the HLA-Cw6 promoter due to the variant kappaB. Cotransfecting NF kappaB1 with RelA further increased activity of the HLA-A2 and -B7, but not HLA Cw6, promoters. All three promoters were activated by MHC class II trans activator, but not CREB-binding protein, whereas IFN regulatory factor-1 and -2 weakly activated the HLA-B7 and -Cw6, but not HLA-A2, promoters. These studies illustrate common and locus-specific mechanisms that may be targeted to modulate immune reactions. PMID- 12574357 TI - Restoration of Ig secretion: mutation of germline-encoded residues in T15L chains leads to secretion of free light chains and assembled antibody complexes bearing secretion-impaired heavy chains. AB - We previously described T15H chain mutants that were impaired in assembly with L chain and in ability to be secreted from the cell. The unmutated T15L chain is unusual in that it is secretion-impaired in the absence of assembly with H chain. The T15L chain preferentially pairs with T15H in vivo, suggesting that if we introduced mutations that would allow secretion of free T15L chain, they might also lead to the secretion of the complex with the defective H chain. We mutated four positions in the germline T15L that had amino acids infrequently found in other kappa-chains. Mutation to the most frequently occurring amino acid at three of the four positions allowed secretion of free L chain, while the combination of two secretion-restoring mutations was synergistic. Coexpression of secretion restored mutant L chains with the secretion-defective mutant H chains rescued secretion of the assembled H(2)L(2) complex, suggesting that during somatic hypermutation in vivo, deleterious mutations at the H chain may be compensated by mutations on the L chain. To our knowledge, this is the first example of mutations in IgL chains that are able to restore secretion-defective H chains to secretion competence in mammalian cells. PMID- 12574358 TI - Structure function differences in nonpeptide CCR1 antagonists for human and mouse CCR1. AB - A useful strategy for identifying ligand binding domains of G protein-coupled receptors has been the exploitation of species differences in antagonist potencies. We have used this approach for the CCR1 chemokine receptor with a novel series of antagonists, the 4-hydroxypiperidines, which were discovered by high throughput screening of human CCR1 and subsequently optimized. The structure activity relationships for a number of different 4-hydroxypiperidine antagonists for human and mouse CCR1 were examined by receptor binding and functional assays. These compounds exhibit major differences in their rank order of potency for the human and mouse chemokine receptor CCR1. For example, the initial lead template, BX 510, which was a highly potent functional antagonist for human CCR1 (K(i) = 21 nM) was >400-fold less active on mouse CCR1 (K(i) = 9150 nM). However, increasing the length of the linker between the piperidine and dibenzothiepine groups by one methylene group generated a compound, BX 511, which was equipotent for both human and mouse CCR1. These and other analogs of the lead template BX 510, which have major differences in potency for human and mouse CCR1, are described, and a model for their interaction with human CCR1 is presented. PMID- 12574359 TI - Crystal structure of a hydrophobic immunodominant antigenic site on hepatitis C virus core protein complexed to monoclonal antibody 19D9D6. AB - The first crystal structure of a complex between a hepatitis C virus (HCV) core protein-derived peptide (residues 13-40) and the Ab fragment of a murine mAb (19D9D6) has been solved, allowing determination of the recognized epitope and elucidation of its conformation. This Ab, raised against the first 120 residues of the core protein, recognizes core particles and strongly competes with anticore human Abs, suggesting that it is highly representative of the human anti HCV core response. Its epitope lies within the first 45 aa of the protein, the major antigenic segment of core recognized both by murine and human Abs. Surprisingly, the recognized epitope (29-37: QIVGGVYLL) has an unusual preponderance of hydrophobic residues, some of which are buried in a small hydrophobic core in the nuclear magnetic resonance structure of the peptide (2 45) in solution, suggesting that the Ab may induce a structural rearrangement upon recognition. The flexibility may reside entirely within the Ag, since the Fab'-peptide complex structure at 2.34 A shows that the Ab binding site is hardly perturbed by complexation. Given that the recognized residues are unlikely to be solvent exposed, we are left with the interesting possibility that Ab-core interactions may take place in a nonaqueous environment. PMID- 12574360 TI - Associations of MHC ancestral haplotypes with resistance/susceptibility to AIDS disease development. AB - We tested the association of MHC ancestral haplotypes with rapid or slow progression to AIDS by comparing their frequencies in the French genetics of resistance/susceptibility to immunodeficiency virus cohort with that reported in a control French population. Seven ancestral haplotypes were identified in the genetics of resistance/susceptibility to immunodeficiency virus cohort with a frequency >1%. The 8.1 (odds ratio (OR) = 3, p = 0.006), 35.1 (OR = 5.7, p = 0.001), and 44.2 (OR = 3.4, p = 0.007) ancestral haplotypes were associated with rapid progression, whereas the 35.2 (OR = 3.6, p = 0.001), 44.1 (OR = 5.4, p < 10(-4)), and 57.1 (OR = 5.8, p < 10(-4)) ancestral haplotypes were associated with slow progression to AIDS. Although the frequency of each ancestral haplotype is low in the population, the OR were quite higher than those previously obtained for single HLA allele associations, with some p values as low as 10(-4). The analysis of the recombinant fragments of these haplotypes allowed the identification of the MHC regions in the 35.1, 35.2, and 44.2 haplotypes associated with rapid progression to AIDS and the MHC regions of the 44.1 and 57.1 haplotypes associated with slow progression to AIDS. Previous studies have identified single HLA alleles associated with disease progression. Our results on recombinant fragments confirm the direct role of HLA-B35 in rapid progression. Associations with HLA-A29 and -B57 might be due to linkage disequilibrium with other causative genes within the MHC region. PMID- 12574361 TI - CTLA-4 in filarial infections: implications for a role in diminished T cell reactivity. AB - To determine the role that CTLA-4 might play in mediating the diminished parasite Ag-specific T cell responsiveness that is characteristically seen in filaria infected patients, several study populations and methods were used. First, quantitative assessment of mRNA expression determined that PBMC from uninfected adolescents exposed in utero to microfilarial (Mf) Ag demonstrated a strong up regulation of CTLA-4 to the Mf stage of the parasite in contrast to that observed in cells from children born of uninfected mothers (p = 0.005). Next, the frequency of CTLA-4 expression was examined using flow cytometry in cells from filaria-infected and -uninfected individuals ex vivo. Individuals born in filarial endemic regions of the world (with long-standing infections) had greater percentages of CD4(+)CTLA-4(+) cells than did expatriate infected or uninfected individuals (p = 0.005 and 0.05, respectively); in addition, Mf(+) patients demonstrated higher frequencies of CD4(+)CTLA-4(+) and CD8(+)CTLA-4(+) cells (p = 0.027 and 0.037, respectively) than did Mf(-) infected individuals. Of interest, the greatest intensity of CTLA-4 expression occurred in CD4(+)CD25(+) cells, a population purported to include suppressor cells. Finally, in vitro blocking of CTLA-4 expression in PBMC from filaria-infected individuals induced a mean increase of 44% in IL-5 production to Mf Ag, whereas there was a concurrent mean decrease of 42% in IFN-gamma production, suggesting that CTLA-4 also acts to alter the Th1/Th2 balance in filaria-infected individuals. Together, these data indicate a significant role for CTLA-4 in regulating the host response to filarial infections and that factors such as length of exposure and patency are important codeterminants. PMID- 12574362 TI - Constrained intracellular survival of Mycobacterium tuberculosis in human dendritic cells. AB - Dendritic cells (DCs) are likely to play a key role in immunity against Mycobacterium tuberculosis, but the fate of the bacterium in these cells is still unknown. Here we report that, unlike macrophages (Mphis), human monocyte-derived DCs are not permissive for the growth of virulent M. tuberculosis H37Rv. Mycobacterial vacuoles are neither acidic nor fused with host cell lysosomes in DCs, in a mode similar to that seen in mycobacterial infection of Mphis. However, uptake of the fluid phase marker dextran, and of transferrin, as well as accumulation of the recycling endosome-specific small GTPase Rab11 onto the mycobacterial phagosome, are almost abolished in infected DCs, but not in Mphis. Moreover, communication between mycobacterial phagosomes and the host-cell biosynthetic pathway is impaired, given that <10% of M. tuberculosis vacuoles in DCs stained for the endoplasmic reticulum-specific proteins Grp78/BiP and calnexin. This correlates with the absence of the fusion factor N-ethylmaleimide sensitive factor onto the vacuolar membrane in this cell type. Trafficking between the vacuoles and the host cell recycling and biosynthetic pathways is strikingly reduced in DCs, which is likely to impair access of intracellular mycobacteria to essential nutrients and may thus explain the absence of mycobacterial growth in this cell type. This unique location of M. tuberculosis in DCs is compatible with their T lymphocyte-stimulating functions, because M. tuberculosis-infected DCs have the ability to specifically induce cytokine production by autologous T lymphocytes from presensitized individuals. DCs have evolved unique subcellular trafficking mechanisms to achieve their Ag-presenting functions when infected by intracellular mycobacteria. PMID- 12574363 TI - The induction and kinetics of antigen-specific CD8 T cells are defined by the stage specificity and compartmentalization of the antigen in murine toxoplasmosis. AB - Toxoplasma gondii forms different life stages, fast-replicating tachyzoites and slow-growing bradyzoites, in mammalian hosts. CD8 T cells are of crucial importance in toxoplasmosis, but it is unknown which parasite stage is recognized by CD8 T cells. To analyze stage-specific CD8 T cell responses, we generated various recombinant Toxoplasma gondii expressing the heterologous Ag beta galactosidase (beta-gal) and studied whether 1) secreted or cytoplasmic Ags and 2) tachyzoites or bradyzoites, which persist intracerebrally, induce CD8 T cells. We monitored the frequencies and kinetics of beta-gal-specific CD8 T cells in infected mice by MHC class I tetramer staining. Upon oral infection of B6C (H 2(bxd)) mice, only beta-gal-secreting tachyzoites induced beta-gal-specific CD8 T cells. However, upon secondary infection of mice that had received a primary infection with tachyzoites secreting beta-gal, beta-gal-secreting tachyzoites and bradyzoites transiently increased the frequency of intracerebral beta-gal specific CD8 T cells. Frequencies of splenic and cerebral beta-gal-specific CD8 T cells peaked at day 23 after infection, thereafter persisting at high levels in the brain but declining in the spleen. Splenic and cerebral beta-gal-specific CD8 T cells produced IFN-gamma and were cytolytic upon specific restimulation. Thus, compartmentalization and stage specificity of an Ag determine the induction of CD8 T cells in toxoplasmosis. PMID- 12574365 TI - The mitochondrial network of human neutrophils: role in chemotaxis, phagocytosis, respiratory burst activation, and commitment to apoptosis. AB - It is commonly assumed that human neutrophils possess few, if any, functional mitochondria and that they do not depend on these organelles for cell function. We have used the fluorescent mitochondrial indicators, JC-1, MitoTracker Red, and dihydrorhodamine 123 to show that live neutrophils possess a complex mitochondrial network that extends through the cytoplasm. The membrane potential of these mitochondria was rapidly (within 2 min) disrupted by the addition of FCCP (IC(50) = 20 nM), but not by the Fo-ATPase inhibitor, oligomycin (at up to 7 microg/ml). However, inhibition of mitochondrial function with both agents resulted in cell shape changes. Neither activation of the respiratory burst nor phagocytosis of either latex particles or serum-opsonized Staphylococcus aureus was affected by the addition of FCCP or oligomycin. However, FCCP inhibited chemotaxis at concentrations that paralleled disruption of mitochondrial membrane potential. Furthermore, prolonged (2-h) incubation with oligomycin resulted in an impaired ability to activate a respiratory burst and also inhibited chemotaxis. These observations indicate that intact mitochondrial function is required to sustain some neutrophil functions, but not for the rapid initiation of the respiratory burst or phagocytosis. Loss of mitochondrial membrane potential was a very early marker for commitment of neutrophils into apoptosis and preceded the appearance of phosphatidylserine on the cell surface. However, inhibition of mitochondrial function did not accelerate the rate of neutrophil apoptosis. These data shed important insights into the hitherto unrecognized importance of mitochondria in the function of neutrophils during infection and inflammation. PMID- 12574364 TI - CD4+ T cells mediate abscess formation in intra-abdominal sepsis by an IL-17 dependent mechanism. AB - Abscess formation associated with intra-abdominal sepsis causes severe morbidity and can be fatal. Previous studies have implicated T cells in the pathogenesis of abscess formation, and we have recently shown that CD4(+) T cells activated in vitro by zwitterionic capsular polysaccharides from abscess-inducing bacteria such as Staphylococcus aureus and Bacteroides fragilis initiate this host response when transferred to naive rats. In this study, we show that mice deficient in alphabetaTCR-bearing T cells or CD4(+) T cells fail to develop abscesses following challenge with B. fragilis or abscess-inducing zwitterionic polysaccharides, compared with CD8(-/-) or wild-type animals. Transfer of CD4(+) T cells from wild-type mice to alphabetaTCR(-/-) animals reconstituted this ability. The induction of abscesses required T cell costimulation via the CD28-B7 pathway, and T cell transfer experiments with STAT4(-/-) and STAT6(-/-) mice demonstrated that this host response is dependent on STAT4 signaling. Significantly higher levels of IL-17, a proinflammatory cytokine produced almost exclusively by activated CD4(+) T cells, were associated with abscess formation in Th2-impaired (STAT6(-/-)) mice, while STAT4(-/-) mice had significantly lower levels of this cytokine than control animals. The formation of abscesses was preceded by an increase in the number of activated CD4(+) T cells in the peritoneal cavity 24 h following bacterial challenge. Confocal laser-scanning microscopy analysis revealed that CD4(+) T cells comprise the abscess wall in these animals and produce IL-17 at this site. Administration of a neutralizing Ab specific for IL-17 prevented abscess formation following bacterial challenge in mice. These data delineate the specific T cell response necessary for the development of intra-abdominal abscesses and underscore the role of IL-17 in this disease process. PMID- 12574366 TI - Mouse SWAM1 and SWAM2 are antibacterial proteins composed of a single whey acidic protein motif. AB - Antibacterial proteins are important participants in the innate immunity system. Elafin and SLPI are the whey acidic protein (WAP) motif proteins with both antibacterial activity and antiprotease activity, and their role in innate immunity is under intense investigation. We cloned two novel antibacterial WAP motif proteins from mice, SWAM1 and SWAM2. SWAM1 and SWAM2 are composed of a signal sequence and a single WAP motif that has high homologies with the WAP motifs of elafin and SLPI. SWAM1 is constitutively expressed in kidney and epididymis, and is induced in the pneumonic lung. SWAM2 is constitutively expressed in tongue. SWAM1 and SWAM2 inhibit the growth of both Escherichia coli and Staphylococcus aureus at a IC(90) (concentration that achieves 90% inhibition) of 10 microM. Human genes LOC149709 and huWAP2 are considered to be human SWAM1 and SWAM2, respectively. These and several WAP motif proteins (WAP1, elafin, SLPI, HE4, eppin, C20orf170, LOC164237, and WFDC3) form a gene cluster on human chromosome 20, suggesting that they may be derived from the same ancestral gene by gene duplication. Our results underscore the role of the WAP motif as a skeletal motif to form antibacterial proteins, and warrant the study of antibacterial activity in other WAP motif proteins. PMID- 12574367 TI - Prevention of spontaneous breast carcinoma by prophylactic vaccination with dendritic/tumor fusion cells. AB - Genetically modified mice with spontaneous development of mammary carcinoma provide a powerful tool to study the efficacy of tumor vaccines, since they mimic breast cancer development in humans. We used a transgenic murine model expressing polyomavirus middle T oncogene and mucin 1 tumor-associated Ag to determine the preventive effect of a dendritic/tumor fusion cell vaccine. The MMT (a transgenic murine model) mice developed mammary carcinoma between the ages of 65-108 days with 100% penetrance. No spontaneous CTL were detected. However, prophylactic vaccination of MMT mice with dendritic/tumor fusion cells induced polyclonal CTL activity against spontaneous mammary carcinoma cells and rendered 57-61% of the mice free of the disease at the end of experiment (180 days). Furthermore, the level of CTL activity was maintained with multiple vaccinations. The antitumor immunity induced by vaccination with dendritic/tumor fusion cells reacted differently to injected tumor cells and autochthonous tumor. Whereas the injected tumor cells were rejected, the autochthonous tumor evaded the attack and was allowed to grow. Collectively these results indicate that prophylactic vaccination with dendritic/tumor fusion cells confers sufficient antitumor immunity to counter the tumorigenesis of potent oncogenic products. The findings in the present study are highly relevant to cancers in humans. PMID- 12574368 TI - CD8+ T cells accumulate in the lungs of Mycobacterium tuberculosis-infected Kb-/ Db-/- mice, but provide minimal protection. AB - Recent studies have shown that MHC class I molecules play an important role in the protective immune response to Mycobacterium tuberculosis infection. Here we showed that mice deficient in MHC class Ia, but possessing MHC class Ib (K(b-/ )D(b-/-) mice), were more susceptible to aerosol infection with M. tuberculosis than control mice, but less susceptible than mice that lack both MHC class Ia and Ib (beta(2)m(-/-) mice). The susceptibility of K(b-/-)D(b-/-) mice cannot be explained by the failure of CD8(+) T cells (presumably MHC class Ib-restricted) to respond to the infection. Although CD8(+) T cells were a relatively small population in uninfected K(b-/-)D(b-/-) mice, most already expressed an activated phenotype. During infection, a large percentage of these cells further changed their cell surface phenotype, accumulated in the lungs at the site of infection, and were capable of rapidly producing IFN-gamma following TCR stimulation. Histopathologic analysis showed widespread inflammation in the lungs of K(b-/ )D(b-/-) mice, with a paucity of lymphocytic aggregates within poorly organized areas of granulomatous inflammation. A similar pattern of granuloma formation has previously been observed in other types of MHC class I-deficient mice, but not CD8alpha(-/-) mice. Thus, neither the presence of MHC class Ib molecules themselves, nor the activity of a population of nonclassical CD8(+) effector cells, fully restored the deficit caused by the absence of MHC class Ia molecules, suggesting a unique role for MHC class Ia molecules in protective immunity against M. tuberculosis. PMID- 12574369 TI - NF-kappa B1 is required for optimal CD4+ Th1 cell development and resistance to Leishmania major. AB - The NF-kappaB family of transcription factors regulates the expression of a wide range of immune response genes involved in immunity to pathogens. However, the need for individual family members in regulating innate and adaptive immune responses in vivo has yet to be clearly defined. We investigated the role of NF kappaB1 in the induction of protective IL-12-dependent Th1 cell responses following infection with the intracellular protozoan parasite Leishmania major. Whereas wild-type C57BL/6 mice controlled parasite replication, NF-kappaB1 knockout (KO) mice were susceptible to infection, developing chronic unresolving lesions associated with persistent parasites. There was a profound defect in Ag specific CD4(+) T cell proliferation and IFN-gamma production in infected KO mice, although innate responses-including IL-12 production and control of intracellular parasite replication by macrophages-were intact. In vitro polyclonal stimulation of purified naive KO T cells revealed an intrinsic defect in CD4(+) T cell proliferation associated with reduced IL-2 receptor expression, but operating independently of APC function and IL-2 production. Critically, the frequency of proliferating KO CD4(+) T cells secreting IFN-gamma matched that of wild-type cells, suggesting that NF-kappaB1 was not required for efficient transcription of the IFN-gamma gene. Taken together, these results identify a novel role for NF-kappaB1 in CD4(+) T cell proliferation and the development of Th1 cell responses required for protective immunity against intracellular pathogens. PMID- 12574370 TI - TNF plays an essential role in tumor regression after adoptive transfer of perforin/IFN-gamma double knockout effector T cells. AB - We have recently shown that effector T cells (T(E)) lacking either perforin or IFN-gamma are highly effective mediators of tumor regression. To rule out compensation by either mechanism, T(E) deficient in both perforin and IFN-gamma (perforin knockout (PKO)/IFN-gamma knockout (GKO)) were generated. The adoptive transfer of PKO/GKO T(E) mediated complete tumor regression and cured wild-type animals with established pulmonary metastases of the B16BL6-D5 (D5) melanoma cell line. PKO/GKO T(E) also mediated tumor regression in D5 tumor-bearing PKO, GKO, or PKO/GKO recipients, although in PKO/GKO recipients efficacy was reduced. PKO/GKO T(E) exhibited tumor-specific TNF-alpha production and cytotoxicity in a 24-h assay, which was blocked by the soluble TNFRII-human IgG fusion protein (TNFRII:Fc). Blocking TNF in vivo by administering soluble TNFR II fusion protein (TNFRII:Fc) significantly reduced the therapeutic efficacy of PKO/GKO, but not wild-type T(E). This study identifies perforin, IFN-gamma, and TNF as a critical triad of effector molecules that characterize therapeutic antitumor T cells. These insights could be used to monitor and potentially tune the immune response to cancer vaccines. PMID- 12574371 TI - Regression of a mammary adenocarcinoma in STAT6-/- mice is dependent on the presence of STAT6-reactive T cells. AB - Polarization of the immune response toward a type 1 cytokine profile has been posited to be associated with a therapeutic antitumor immune response. STAT6-/- mice are unable to generate a type 2 immune response, and instead mount an enhanced type 1 response. STAT6-/- mice are significantly more resistant to 4T1, a mammary adenocarcinoma cell line, resisting a 10-fold higher tumor dose compared with wild-type (wt) BALB/c mice. An analysis of the T cells from tumor bearing STAT6-/- mice revealed that they contained a population primed by a peptide (STAT6(531-539)) of the STAT6 protein expressed in 4T1. The adoptive transfer of T cells from STAT6(531-539)-vaccinated STAT6-/- mice significantly reduced the number of 4T1 pulmonary metastases in recipient mice. Additionally, the role of these STAT6(531-539)-reactive T cells against s.c. 4T1 tumor challenge was determined by tumor-challenging wt BALB/c mice reconstituted with STAT6-/- bone marrow, thereby assessing whether a polarized type 1 immune response in the absence of STAT6-reactive T cells was sufficient to reject a 4T1 tumor challenge. T cells from the STAT6-/- bone marrow chimeras failed to recognize the STAT6(531-539), and these mice proved to be as susceptible as wt BALB/c mice to 4T1 challenge. This demonstrated that the absence of STAT6(531 539)-reactive T cells correlated with the inability to reject 4T1 challenge. Additionally, these data emphasize that the enhanced ability to mount a type 1 polarized immune response is inconsequential if a sufficient antitumor immune response is not primed by the tumor. PMID- 12574372 TI - Memory inflation: continuous accumulation of antiviral CD8+ T cells over time. AB - CD8+ T lymphocytes play an important role in the control of intracellular pathogens during both acute and persistent infections. This is particularly true in the case of persistent herpesviruses such as human CMV, which are typified by large virus-specific CD8+ T cell populations during viral latency. To understand the origin of these populations and the factors shaping them over time, we investigated the CD8+ T cell response after murine CMV (MCMV) infection. The kinetics of the acute response were characterized by rapid expansion of activated T cells, followed by a contraction phase. Thereafter, we observed a striking pattern, where MCMV-specific memory CD8+ T cells steadily accumulated over time, with 20% of all CD8+ T cells at 1 year specific for one MCMV epitope. Accumulation of MCMV-specific CD8+ T lymphocytes was seen in all organs tested and was associated with continuous activation of specific CD8+ T lymphocytes, primarily within lymph nodes. The pattern of accumulation was observed in only two of five epitopes tested, and was accompanied by a gradual restriction in usage of the variable region of the TCR beta-chain over time. This novel pattern of a virus-specific CD8+ T cell response suggests that continuous or repetitive exposure to Ag can slowly mold memory T cell populations over time. This may be relevant for understanding the evolution of the large human CMV-specific CD8+ T cell populations seen in humans. PMID- 12574373 TI - Modulation of HLA-A*0201-restricted T cell responses by natural polymorphism in the IE1(315-324) epitope of human cytomegalovirus. AB - Cytotoxic T lymphocytes play a central role in the control of persistent human CMV (HCMV) infection and reactivation. In healthy virus carriers, the specific CD8(+) CTL response is almost entirely directed against the virion tegument protein pp65 and/or the 72-kDa major immediate early protein, IE1. Studies that included a large panel of HCMV(+) donors suggested that immunorelevance of pp65 and IE1 was directly related with individual HLA haplotype difference. Nevertheless, there are no data on the incidence of HCMV natural polymorphism on virus-specific CTL responses. To assess the impact of IE1 polymorphism on CTL response, we have sequenced in 103 clinical isolates the DNA region corresponding to IE1(315-324), an immunodominant epitope presented by HLA-A*0201 molecules. Seven peptidic variants were found with extensive difference in their frequencies. The response of four HLA-A*0201-restricted anti-IE1 T lymphocyte clones, which were previously generated from one donor against autologous B lymphoblastoid cells expressing a recombinant clinical variant of IE1, was then evaluated using target cells loaded with mutant synthetic peptides or expressing rIE1 variants. One of four clones, which have been sorted 19 times among 22 clones targeted against IE1(315-324), recognized six of the seven tested variant epitopes. All three other clones showed distinct reactivity patterns to target cells loaded with the different mutant peptides or expressing IE1 variants. Therefore, in the HLA-A2 context, clonal expansions of anti-IE1 memory CTLs may confer a protection against HCMV successive infections and reactivations by killing cells presenting most of the naturally occurring IE1(315-324) epitope variants. PMID- 12574374 TI - IL-13 is sufficient for respiratory syncytial virus G glycoprotein-induced eosinophilia after respiratory syncytial virus challenge. AB - Although well studied in settings of helminth infection and allergen sensitization, the combined contributions of IL-4 and IL-13 and their signaling pathways in models of viral pathogenesis have not been reported. Using a murine model of respiratory syncytial virus (RSV) infection, we evaluated the contribution of IL-13, alone and in conjunction with IL-4, during immunization with recombinant vaccinia virus expressing RSV G glycoprotein (vvGs) or with formalin-inactivated RSV (FI-RSV). We showed that both IL-4 and IL-13 activity must be inhibited to modulate G-specific responses resulting in severe RSV induced disease. Inhibition of IL-4 or IL-13 activity alone had minimal impact on disease in vvGs-immunized mice. However, treatment of IL-4-deficient mice with IL 13Ra during vvGs immunization reduced IL-5, IL-13, and eotaxin production and pulmonary eosinophilia after RSV challenge. In contrast, FI-RSV-induced immune responses were diminished when either IL-4 or IL-13 activity was blocked. After RSV challenge, these type 2 T cell responses were also diminished in vvGs-primed IL-4Ralpha-deficient mice. Our data suggest that secreted vvGs uses mechanisms requiring signaling through the IL-4Ralpha-chain by either IL-4 or IL-13 for induction of eosinophilia and is the first description of the relative contributions of IL-4, IL-13, and their receptors in viral pathogenesis. PMID- 12574375 TI - Persistence and turnover of antigen-specific CD4 T cells during chronic tuberculosis infection in the mouse. AB - CD4 T cells are critical for resistance to Mycobacterium tuberculosis infection, but how effective T cell responses are maintained during chronic infection is not well understood. To address this question we examined the CD4 T cell response to a peptide from ESAT-6 during tuberculosis infection in the mouse. The ESAT-6(1 20)/IA(b)-specific CD4 T cell response in the lungs, mediastinal lymph nodes, and spleen reached maxima 3-4 wk postinfection, when the bacteria came under the control of the immune response. Once chronic infection was established, the relative frequencies of Ag-specific CD4 T cells were maintained at nearly constant levels for at least 160 days. ESAT-6(1-20)/IA(b)-specific CD4 T cells that responded in vitro expressed activation markers characteristic of chronically activated effector cells and used a limited Vbeta repertoire that was clonally stable in vivo for at least 12 wk. 5-Bromo-2-deoxyuridine incorporation studies indicated a relatively high rate of cell division among both total CD4 and ESAT-6(1-20)/IA(b)-specific CD4 T cells during acute infection, but the degree of 5-bromo-2-deoxyuridine incorporation by both the CD4 T cells and the Ag specific cells declined at least 3-fold during chronic infection. The data indicate that the peripheral ESAT-6(1-20)/IA(b)-specific CD4 T cell response to M. tuberculosis is characterized during the acute phase of infection by a period of extensive proliferation, but once bacterial control is achieved, this is followed during chronic infection by an extended containment phase that is associated with a persistent response of activated, yet more slowly proliferating, T cells. PMID- 12574376 TI - Role of CD4 T cell help and costimulation in CD8 T cell responses during Listeria monocytogenes infection. AB - CD4 T cells are known to assist the CD8 T cell response by activating APC via CD40-CD40 ligand (L) interactions. However, recent data have shown that bacterial products can directly activate APC through Toll-like receptors, resulting in up regulation of costimulatory molecules necessary for the efficient priming of naive T cells. It remains unclear what role CD4 T cell help and various costimulation pathways play in the development of CD8 T cell responses during bacterial infection. In this study, we examined these questions using an intracellular bacterium, Listeria monocytogenes, as a model of infection. In CD4 T cell-depleted, CD4(-/-), and MHC class II(-/-) mice, L. monocytogenes infection induced CD8 T cell activation and primed epitope-specific CD8 T cells to levels commensurate with those in normal C57BL/6 mice. Furthermore, these epitope specific CD8 T cells established long-term memory in CD4(-/-) mice that was capable of mounting a protective recall response. In vitro analysis showed that L. monocytogenes directly stimulated the activation and maturation of murine dendritic cells. The CD8 T cell response to L. monocytogenes was normal in CD40L( /-) mice but defective in CD28(-/-) and CD137L(-/-) mice. These data show that in situations where infectious agents or immunogens can directly activate APC, CD8 T cell responses are less dependent on CD4 T cell help via the CD40-CD40L pathway but involve costimulation through CD137-CD137L and B7-CD28 interactions. PMID- 12574377 TI - Expression of the long form of human FLIP by retroviral gene transfer of hemopoietic stem cells exacerbates experimental autoimmune encephalomyelitis. AB - Subsidence of inflammation and clinical recovery in experimental autoimmune encephalomyelitis (EAE) is postulated to involve apoptosis of inflammatory cells. To test this concept, we examined the effects of overexpressing the long form of human FLICE-inhibitory protein, a potent inhibitor of death receptor-mediated apoptosis, in myelin oligodendrocyte glycoprotein-induced EAE in DBA/1 mice. We found that overexpression of the long form of human FLICE-inhibitory protein by retroviral gene transfer of hemopoietic stem cells led to a clinically more severe EAE in these mice compared with control mice receiving the retroviral vector alone. The exacerbated disease was evident by an enhanced and prolonged inflammatory reaction in the CNS of these animals compared with control mice. The acute phase of EAE was characterized by a massive infiltration of macrophages and granulocytes and a simultaneous increase in TNF-alpha production in the CNS. In the chronic phase of the disease, there was a prolonged inflammatory response in the form of persistent CD4(+) T and B cells in the CNS and a peripheral Th1 cytokine bias caused by elevated levels of IFN-gamma and reduced levels of IL-4 in the spleen. Our findings demonstrate that death receptor-mediated apoptosis can be important in the pathogenesis of EAE and further emphasize the need for effective apoptotic elimination of inflammatory cells to achieve disease remission. PMID- 12574378 TI - Suppression of NF-kappa B activation and proinflammatory cytokine expression by Shiga toxin-producing Escherichia coli. AB - The NF-kappaB family of transcription factors forms one of the first lines of defense against infectious disease by inducing the expression of genes involved in inflammatory and immune responses. In this study, we analyzed the impact of Shiga toxin-producing Escherichia coli (STEC) on the NF-kappaB DNA-binding activity in HeLa cells. After a period of weak initial activation, DNA binding of NF-kappaB was actively suppressed by viable, E. coli secreted protein B (EspB) secreting STEC. Sustained NF-kappaB activity was observed either using an isogenic mutant lacking EspB or after gentamicin-based killing of STEC after allowing bacterial attachment. These observations indicate that the ability of STEC to cause NF-kappaB activation is suppressed by a translocated bacterial effector protein, which is either EspB itself or requires EspB for delivery into the host cell. We found that STEC, enterohemorrhagic E. coli, and enteropathogenic E. coli all interfere with NF-kappaB activation initiated by TNF alpha, indicating that suppression of signal-induced NF-kappaB activity is a property common to several attaching and effacing bacteria. As a consequence of NF-kappaB suppression, wild-type STEC induces significantly lower mRNA levels of IL-8, IL-6, and IL-1alpha upon prolonged infection periods compared with bacteria lacking EspB. For IL-8 and IL-6, the suppressive effect was also reflected at the level of cytokine secretion. Suppression of both basal and signal-induced NF kappaB DNA binding by attaching and effacing-inducing bacteria appears to be an active strategy to counteract host defense responses, thus favoring intestinal colonization by these pathogens. PMID- 12574379 TI - Dual roles of IL-4 in lung injury and fibrosis. AB - Increased lung IL-4 expression in pulmonary fibrosis suggests a potential pathogenetic role for this cytokine. To dissect this role, bleomycin-induced pulmonary inflammation and fibrosis were analyzed and compared in wild type (IL 4(+/+)) vs IL-4-deficient (IL-4(-/-)) mice. Lethal pulmonary injury after bleomycin treatment was higher in IL-4(-/-) vs IL-4(+/+) mice. By administration of anti-CD3 Abs, we demonstrated that this early response was linked to the marked T lymphocyte lung infiltration and to the overproduction of the proinflammatory mediators such as TNF-alpha, IFN-gamma, and NO in IL-4(-/-) mice. In contrast to this early anti-inflammatory/immunosuppressive role, during later stages of fibrosis, IL-4 played a profibrotic role since IL-4(-/-) mice developed significantly less pulmonary fibrosis relative to IL-4(+/+) mice. However, IL-4 failed to directly stimulate proliferation, alpha-smooth muscle actin, and type I collagen expression in lung fibroblasts isolated from the wild-type mice. Upon appropriate stimulation with other known fibrogenic cytokines, fibroblasts from IL-4(-/-) mice were relatively deficient in the studied parameters in comparison to fibroblasts isolated from IL-4(+/+) mice. Taken together, these data suggest dual effects of IL-4 in this model of lung fibrosis: 1) limiting early recruitment of T lymphocytes, and 2) stimulation of fibrosis chronically. PMID- 12574380 TI - Akt as a mediator of secretory phospholipase A2 receptor-involved inducible nitric oxide synthase expression. AB - The induction of inducible NO synthase (iNOS) by group IIA phospholipase A(2) (PLA(2)) involves the stimulation of a novel signaling cascade. In this study, we demonstrate that group IIA PLA(2) up-regulates the expression of iNOS through a novel pathway that includes M-type secretory PLA(2) receptor (sPLA(2)R), phosphatidylinositol 3-kinase (PI3K), and Akt. Group IIA PLA(2) stimulated iNOS expression and promoted nitrite production in a dose- and time-dependent manner in Raw264.7 cells. Upon treating with group IIA PLA(2), Akt is phosphorylated in a PI3K-dependent manner. Pretreatment with LY294002, a PI3K inhibitor, strongly suppressed group IIA PLA(2)-induced iNOS expression and PI3K/Akt activation. The promoter activity of iNOS was stimulated by group IIA PLA(2), and this was suppressed by LY294002. Transfection with Akt cDNA resulted in Akt protein overexpression in Raw264.7 cells and effectively enhanced the group IIA PLA(2) induced reporter activity of the iNOS promoter. M-type sPLA(2)R was highly expressed in Raw264.7 cells. Overexpression of M-type sPLA(2)R enhanced group IIA PLA(2)-induced promoter activity and iNOS protein expression, and these effects were abolished by LY294002. However, site-directed mutation in residue responsible for PLA(2) catalytic activity markedly reduced their ability to production of nitrites and expression of iNOS. These results suggest that group IIA PLA(2) induces nitrite production by involving of M-type sPLA(2)R, which then mediates signal transduction events that lead to PI3K/Akt activation. PMID- 12574381 TI - Mechanisms of inhibition of collagen-induced arthritis by murine IL-18 binding protein. AB - IL-18 is an important cytokine in autoimmune and inflammatory diseases through the induction of IFN-gamma, TNF-alpha, and IL-1. We report herein that collagen induced arthritis (CIA) in mice is inhibited by treatment with murine IL-18 binding protein (mIL-18BP). CIA was induced in DBA/1J mice by the injection of bovine type II collagen (CII) in IFA with added Mycobacterium tuberculosis on days 0 and 21. The mice were then treated for 3 wk with PBS or with two doses of mIL-18BP (0.5 and 3 mg/kg) as a fusion protein with the Fc portion of murine IgG1. Both the clinical disease activity scores and the histological scores of joint damage were reduced 50% in mice treated with either dose of mIL-18BP. Proliferation of CII-stimulated spleen and lymph node cells as well as the change in serum levels of IgG1 and IgG2a Ab to collagen between days 21 and 42 were decreased in mice treated with mIL-18BP. The production of IFN-gamma, TNF-alpha, and IL-1beta in cultured spleen cells was reduced by in vivo treatment with low dose, but not high dose, mIL-18BP. FACS analysis showed a slight decrease in NK cells and an increase in CD4(+) T cells in spleens of mice treated with mIL-18BP. The steady state mRNA levels of IFN-gamma, TNF-alpha, and IL-1beta in isolated joints were all decreased in mice treated with both doses of mIL-18BP. The mechanisms of mIL-18BP inhibition of CIA include reductions in cell-mediated and humoral immunity to collagen as well as decreases in production of proinflammatory cytokines in the spleen and joints. PMID- 12574382 TI - IL-17, produced by lymphocytes and neutrophils, is necessary for lipopolysaccharide-induced airway neutrophilia: IL-15 as a possible trigger. AB - IL-17 is a cytokine implicated in the regulation of inflammation. We investigated the role of this cytokine in neutrophil recruitment using a model of LPS-induced lung inflammation in mice. In the bronchoalveolar lavage, LPS induced a first influx of neutrophils peaking at day 1, followed by a second wave, peaking at day 2. IL-17 levels were increased during the late phase neutrophilia (day 2), and this was concomitant with an increased number of T cells and macrophages, together with an increase of KC and macrophage-inflammatory protein-2 levels in the lung tissue. Intranasal treatment with a neutralizing murine anti-IL-17 Ab inhibited the late phase neutrophilia. In the bronchoalveolar lavage cells, IL-17 mRNA was detected at days 1, 2, and 3 postchallenge, with a strong expression at day 2. This expression was associated with CD4(+) and CD8(+) cells, but also with neutrophils. When challenged with LPS, despite the absence of T cells, SCID mice also developed a neutrophilic response associated with IL-17 production. In BALB/c mice, IL-15 mRNA, associated mainly with neutrophils, was evidenced 1 day after LPS challenge. In vitro, IL-15 was able to induce IL-17 release from purified spleen CD4(+) cells, but not spleen CD8(+) or airway neutrophils. We have shown that IL-17, produced mainly by CD4(+) cells, but also by neutrophils, plays a role in the mobilization of lung neutrophils following bacterial challenge. In addition, our results suggest that IL-15 could represent a physiological trigger that leads to IL-17 production following bacterial infection. PMID- 12574383 TI - Poly(ADP-ribose) polymerase-1 regulates activation of activator protein-1 in murine fibroblasts. AB - Poly(ADP-ribose) polymerase (PARP)-1 is activated in response to DNA injury in the nucleus of eukaryotic cells and has been implicated in cell dysfunction in inflammation. We investigated the role of PARP-1 on the AP-1 pathway, which is involved in the signal transduction of the inflammatory process. In murine wild type fibroblasts, oxidative challenge by peroxynitrite and hydrogen peroxide or immunological challenge by IL-1 and 20% FCS induced phosphorylation of the mitogen-activated protein kinase kinase-4, activation of c-Jun N-terminal kinase (JNK), and DNA binding of AP-1. In comparative experiments, peroxynitrite induced DNA binding of heat shock factor-1. Pretreatment of wild-type cells with 5-iodo-6 amino-1,2-benzopyrone, a PARP-1 inhibitor, inhibited JNK activation and DNA binding of AP-1. In parallel experiments in PARP-1-deficient fibroblasts, DNA binding of AP-1 was completely abolished. Activation of JNK was significantly elevated at basal condition, but it exhibited a lesser increase after oxidative or immunological challenge than in wild-type fibroblasts. Nuclear content of phosphorylated mitogen-activated protein kinase kinase-4 was observed in PARP-1 deficient cells after peroxynitrite challenge only. Western blotting analysis for AP-1 subunits indicated that c-Fos was similarly expressed in wild-type and PARP 1-deficient cells. Phosphorylated c-Jun was expressed after oxidative or immunological challenge, but not in basal condition, in wild-type cells; however, it was significantly elevated at basal condition and further enhanced after oxidative or immunological challenge in PARP-1-deficient cells. No DNA binding of heat shock factor-1 was observed in PARP-1-deficient cells. These data demonstrate that PARP-1 plays a pivotal role in the modulation of transcription. PMID- 12574384 TI - Lipopolysaccharide down-regulates the leukotriene C4 synthase gene in the monocyte-like cell line, THP-1. AB - We studied the effects of LPS on cysteinyl leukotriene (LT) synthesis and LTC(4) synthase expression in mononuclear phagocytes. Conditioning of the monocyte-like cell line, THP-1, with LPS for 7 days resulted in significantly decreased ionophore-stimulated LTC(4) release. The putative LPS receptor, Toll-like receptor 4, was expressed in THP-1 cells. LPS down-regulated LTC(4) synthase mRNA in THP-1 cells in a dose- and time-dependent manner, with down-regulation observed as early as 4 h. Conditioning of actinomycin D-treated cells with LPS resulted in no change in the rate of LTC(4) synthase mRNA decay. LPS treatment of THP-1 cells, transiently transfected with a LTC(4) synthase promoter (1.35 kb) reporter construct, decreased promoter activity. Neutralization of TNF-alpha and inhibition of mitogen-activated protein kinase kinase/extracellular signal regulated kinase did not inhibit the effect of LPS. Treatment of cells with a Toll-like receptor 4-blocking Ab and an inhibitor of NF-kappaB activation resulted in inhibition of the LPS effect, while activation of NF-kappaB and p50/p65 overexpression down-regulated the LTC(4) synthase gene. LPS down regulates cysteinyl LT release and LTC(4) synthase gene expression in mononuclear phagocytes by an NF-kappaB-mediated mechanism. PMID- 12574385 TI - Parenchymal, but not leukocyte, TNF receptor 2 mediates T cell-dependent hepatitis in mice. AB - TNF-alpha is a central mediator of T cell activation-induced hepatitis in mice, e.g., induced by Pseudomonas exotoxin A (PEA). In this in vivo mouse model of T cell-dependent hepatitis, liver injury depends on both TNFRs. Whereas TNFR1 can directly mediate hepatocyte death, the in vivo functions of TNFR2 in pathophysiology remained unclear. TNFR2 has been implicated in deleterious leukocyte activation in a transgenic mouse model and in enhancement of TNFR1 mediated cell death in cell lines. In this study, we clarify the role of hepatocyte- vs leukocyte-expressed TNFR2 in T cell-dependent liver injury in vivo, using the PEA-induced hepatitis model. Several types of TNFR2-expressing leukocytes, especially neutrophils and NK cells, accumulated within the liver throughout the pathogenic process. Surprisingly, only parenchymal TNFR2 expression, but not the TNFR2 expression on leukocytes, contributed to PEA induced hepatitis, as shown by analysis of wild-type --> tnfr2 degrees and the reciprocal mouse bone marrow chimeras. Furthermore, PEA induced NF-kappaB activation and cytokine production in the livers of both wild-type and tnfr2 degrees mice, whereas only primary mouse hepatocytes from wild-type, but not from tnfr2 degrees, mice were susceptible to cell death induced by a combination of agonistic anti-TNFR1 and anti-TNFR2 Abs. Our results suggest that parenchymal, but not leukocyte, TNFR2 mediates T cell-dependent hepatitis in vivo. The activation of leukocytes does not appear to be disturbed by the absence of TNFR2. PMID- 12574386 TI - Absence of CCR8 does not impair the response to ovalbumin-induced allergic airway disease. AB - Interaction of chemokines with their specific receptors results in tight control of leukocyte migration and positioning. CCR8 is a chemokine receptor expressed mainly in CD4(+) single-positive thymocytes and Th2 cells. We generated CCR8 deficient mice (CCR8(-/-)) to study the in vivo role of this receptor, and describe in this study the CCR8(-/-) mouse response in OVA-induced allergic airway disease using several models, including an adoptive transfer model and receptor-blocking experiments. All CCR8(-/-) mice developed a pathological response similar to that of wild-type animals with respect to bronchoalveolar lavage cell composition, peripheral blood and bone marrow eosinophilia, lung infiltrates, and Th2 cytokine levels in lung and serum. The results contrast with a recent report using one of the OVA-induced asthma models studied here. Similar immune responses were also observed in CCR8(-/-) and wild-type animals in a different model of ragweed allergen-induced peritoneal eosinophilic inflammation, with an equivalent number of eosinophils and analogous increased levels of Th2 cytokines in peritoneum and peripheral blood. Our results show that allergic diseases course without critical CCR8 participation, and suggest that further work is needed to unravel the in vivo role of CCR8 in Th2-mediated pathologies. PMID- 12574387 TI - Synoviocyte-derived CXCL12 is displayed on endothelium and induces angiogenesis in rheumatoid arthritis. AB - CXCL12 (stromal cell-derived factor-1) is a potent CXC chemokine that is constitutively expressed by stromal resident cells. Although it is considered a homeostatic rather than an inflammatory chemokine, CXCL12 has been immunodetected in different inflammatory diseases, but also in normal tissues, ant its potential functions and regulation in inflammation are not well known. In this study, we examined the cellular sources of CXCL12 gene expression and the mechanism and effects of its interactions with endothelial cells in rheumatoid arthritis synovium. We show that CXCL12 mRNA was not overexpressed nor induced in cultured rheumatoid synoviocytes, but it specifically accumulated in the rheumatoid hyperplastic lining layer and endothelium. CXCL12 gene expression was restricted to fibroblast-like synoviocytes, whereas endothelial cells did not express CXCL12 mRNA, but displayed the protein on heparitinase-sensitive factors. CXCL12 colocalized with the angiogenesis marker alpha(v)beta(3) integrin in rheumatoid endothelium and induced angiogenesis in s.c. Matrigel plugs in mice. The angiogenic activity of rheumatoid synovial fluid in vivo was abrogated by specific immunodepletion of CXCL12. Our results indicate that synoviocyte-derived CXCL12 accumulates and it is immobilized on heparan sulfate molecules of endothelial cells, where it can promote angiogenesis and inflammatory cell infiltration, supporting a multifaceted function for this chemokine in the pathogenesis of rheumatoid arthritis. PMID- 12574388 TI - Induction of experimental autoimmune encephalomyelitis in IL-12 receptor-beta 2 deficient mice: IL-12 responsiveness is not required in the pathogenesis of inflammatory demyelination in the central nervous system. AB - IL-12 is thought to be involved in the susceptibility to experimental autoimmune encephalomyelitis (EAE), a Th1 cell-mediated autoimmune disorder of the CNS. IL 12 signals through a heterodimeric receptor (IL-12Rbeta1/IL-12Rbeta2), whose beta2-chain is up-regulated on activated, autoreactive Th1 cells. Contrary to the expectation that the absence of IL-12Rbeta2 would protect from EAE, we found that IL-12Rbeta2-deficient mice developed earlier and more severe disease, with extensive demyelination and CNS inflammation. The inflammatory cells were mainly comprised of CD4(+) T cells, monocyte/macrophages, and dendritic cells. Compared to wild-type mice, IL-12Rbeta2-deficient mice exhibited significantly increased autoantigen-induced proliferative response and increased production of TNF-alpha, GM-CSF, IL-17, IL-18/IL-18Ralpha, and NO. In addition, we found significantly increased levels of IL-23p19 mRNA expression in spleen cells from immunized IL 12Rbeta2(-/-) mice compared with wild-type mice. These findings indicate that IL 12 responsiveness is not required in the pathogenesis of inflammatory demyelination in the CNS, and that, in the absence of IL-12Rbeta2, increased IL 23 and other inflammatory molecules may be responsible for increased severity of EAE. PMID- 12574389 TI - Survival and tumor localization of adoptively transferred Melan-A-specific T cells in melanoma patients. AB - Adoptive T cell therapy has been successfully used for treatment of viral and malignant diseases. However, little is known about the fate and trafficking of transferred Ag-specific T cells. Using the tetramer (TM) technology which allows for detection and quantification of Ag-specific CTL, we assessed the frequency of circulating Melan-A-specific CTL in advanced melanoma patients during adoptive T cell therapy. Melan-A-specific CTL were generated from HLA-A2.1(+) patients by in vitro stimulation of CD8(+) T cells with dendritic cells pulsed with a mutated HLA-A2-binding Melan-A (ELAGIGILTV) peptide. Eight patients received three infusions of 0.25-11 x 10(8) Melan-A-specific CTL i.v. at 2-wk intervals along with low-dose IL-2. The transferred T cell product contained a mean of 42.1% Melan-A-TM(+) CTL. Before therapy, the frequencies of Melan-A-specific CTL in patients' circulating CD8(+) T cells ranged from 0.01 to 0.07%. Characterization of the TM frequencies before and at different time points after transfer revealed an increase of circulating Melan-A-specific CTL up to 2%, correlating well with the number of transferred CTL. An elevated frequency of TM(+) T cells was demonstrated up to 14 days after transfer, suggesting long-term survival and/or proliferation of transferred CTL. Combining TM analysis with a flow cytometry based cytokine secretion assay, unimpaired production of IFN-gamma was demonstrated in vivo for at least 24 h after transfer. Indium-111 labeling of Melan-A-specific CTL demonstrated localization of transferred CTL to metastatic sites as early as 48 h after injection. Overall, the results suggest that in vitro-generated Melan-A-specific CTL survive intact in vivo for several weeks and localize preferentially to tumor. PMID- 12574390 TI - Induction of pemphigus phenotype by a mouse monoclonal antibody against the amino terminal adhesive interface of desmoglein 3. AB - Pemphigus vulgaris (PV) is a life-threatening autoimmune blistering disease that is caused by IgG autoantibodies against the cadherin-type adhesion molecule desmoglein (Dsg)3. Previously, we have generated an active mouse model for PV by adoptive transfer of Dsg3(-/-) splenocytes. In this study, we isolated eight AK series, anti-Dsg3 IgG mAbs from the PV mouse model, and examined their pathogenic activities in induction of blister formation. Intraperitoneal inoculation of the AK23 hybridoma, but not the other AK hybridomas, induced the virtually identical phenotype to that of PV model mice or Dsg3(-/-) mice with typical histology of PV. Epitope mapping with domain-swapped and point-mutated Dsg1/Dsg3 molecules revealed that AK23 recognized a calcium-dependent conformational epitope on Dsg3, which consisted of the V3, K7, P8, and D59 Dsg3-specific residues that formed the adhesive interface between juxtaposed Dsg, as predicted by the crystal structure. The epitopes of the mAbs that failed to show apparent pathogenic activity were mapped in the middle to carboxyl-terminal extracellular region of Dsg3, where no direct intermolecular interaction was predicted. These findings demonstrate the pathogenic heterogeneity among anti-Dsg3 IgG Abs due to their epitopes, and suggest the direct inhibition of adhesive interaction of Dsg as an initial molecular event of blister formation in pemphigus. PMID- 12574391 TI - LF 15-0195 inhibits the development of rat central nervous system autoimmunity by inducing long-lasting tolerance in autoreactive CD4 T cells. AB - Experimental autoimmune encephalomyelitis (EAE) is a T cell-dependent autoimmune disease induced in susceptible animals by a single immunization with myelin basic protein (MBP). LF 15-0195 is a novel immunosuppressor that has been shown to have a potent immunosuppressive effect in several pathological manifestations. The purpose of this study was to investigate the effect of this drug on the induction and progression of established rat EAE and to dissect the mechanisms involved. We show that LF 15-0195 administration at the time of MBP immunization reduces the incidence and severity of EAE in Lewis rats. This drug also inhibits ongoing and passively induced EAE, indicating that LF 15-0195 affects already differentiated pathogenic lymphocytes. Compared with lymph node cells from untreated rats, lymphocytes from MBP-immunized rats treated with LF 15-0195 proliferated equally well in response to MBP in vitro, while their ability to produce effector cytokines and to transfer EAE into syngeneic recipients was significantly reduced. This phenomenon is stable and long-lasting. Indeed, neither IL-12 nor repeated stimulation with naive APC and MBP in vitro rendered MBP-specific CD4 T cells from protected rats encephalitogenic. In conclusion, LF 15-0195 treatment suppresses EAE by interfering with both the differentiation and effector functions of autoantigen-specific CD4 T cells. PMID- 12574392 TI - Peptide fine specificity of anti-glycoprotein 100 CTL is preserved following transfer of engineered TCR alpha beta genes into primary human T lymphocytes. AB - TCR with known antitumor reactivity can be genetically introduced into primary human T lymphocytes and provide promising tools for immunogene therapy of tumors. We molecularly characterized two distinct TCRs specific for the same HLA-A2 restricted peptide derived from the melanocyte differentiation Ag gp100, yet exhibiting different stringencies in peptide requirements. The existence of these two distinct gp100-specific TCRs allowed us to study the preservation of peptide fine specificity of native TCRalphabeta when engineered for TCR gene transfer into human T lymphocytes. Retroviral transduction of primary human T lymphocytes with either one of the two sets of TCRalphabeta constructs enabled T lymphocytes to specifically kill and produce TNF-alpha when triggered by native gp100(pos)/HLA-A2(pos) tumor target cells as well as gp100 peptide-loaded HLA A2(pos) tumor cells. Peptide titration studies revealed that the cytolytic efficiencies of the T lymphocyte transductants were in the same range as those of the parental CTL clones. Moreover, primary human T lymphocytes expressing either one of the two engineered gp100-specific TCRs show cytolytic activities in response to a large panel of peptide mutants that are identical with those of the parental CTL. The finding that two gp100-specific TCR, derived from two different CTL, can be functionally introduced into primary human T lymphocytes without loss of the Ag reactivity and peptide fine specificity, holds great promise for the application of TCR gene transfer in cancer immunotherapy. PMID- 12574393 TI - Absence of IL-4, and not suppression of the Th2 response, prevents development of experimental autoimmune Graves' disease. AB - In autoimmune Graves' disease (GD), autoantibodies bind to the thyrotropin receptor (TSHR) and cause hyperthyroidism. We studied the effects of fms-like tyrosine kinase receptor 3 ligand (Flt3-L) or GM-CSF treatment on the development of experimental autoimmune GD (EAGD) in mice, a slowly progressing Ab-mediated organ-specific autoimmune disease of the thyroid induced by immunization with syngeneic cells expressing TSHR. Flt3-L and GM-CSF treatment resulted in up regulation of CD8a(+) and CD8a(-) dendritic cells, and skewing of cytokine and immune responses to TSHR in favor of Th1 and Th2, respectively. However, this skewing did not persist until the later stages, and thus failed to affect the course or severity of the disease. To determine whether the total absence of either IL-4 or IFN-gamma could affect the development of EAGD, we immunized wild type, IFN-gamma(-/-) and IL-4(-/-) BALB/c mice with TSHR. Nearly 100% of the wild type and IFN-gamma(-/-) mice developed EAGD with optimal TSHR-specific immune responses, while IL-4(-/-) mice completely resisted disease and showed delayed and suboptimal pathogenic Ab response. These data demonstrated that skewing immune responses to TSHR, using either Flt3-L or GM-CSF, in favor of Th1 or Th2, respectively, may not be sufficient to alter the course of the disease, while the complete absence of IL-4, but not IFN-gamma, can prevent the development of EAGD. PMID- 12574394 TI - Essential roles of perforin in antigen-specific cytotoxicity mediated by human CD4+ T lymphocytes: analysis using the combination of hereditary perforin deficient effector cells and Fas-deficient target cells. AB - Although the cytotoxic mechanisms of murine CTLs have been investigated extensively using various mutant and knockout mice, those of human CTLs, especially CD4+ CTLs, are still obscure. To clarify the roles of perforin in Ag specific cytotoxicity mediated by human CD4+ CTLs, alloantigen-specific and HSV specific human CD4+ T lymphocyte bulk lines and clones were established from a patient with hereditary perforin deficiency and her healthy father, and their cytotoxic activities were investigated. Alloantigen-specific CD4+ T lymphocytes expressing perforin exerted cytotoxicity against Fas-negative as well as Fas positive allogeneic B lymphoblastoid cell lines established from members of a family with hereditary Fas deficiency. Perforin-deficient, but not perforin expressing, CD4+ T lymphocytes failed to show strong cytotoxicity against HSV infected autologous B lymphoblastoid cells. Perforin-deficient CD4+ T lymphocytes could exert relatively low level cytotoxicity against allogeneic IFN-gamma treated keratinocytes. Although cytotoxicity mediated by perforin-expressing CD4+ CTLs was almost completely inhibited by concanamycin A, a potent inhibitor of the perforin-mediated cytotoxic pathway, cytotoxicity against IFN-gamma-treated keratinocytes mediated by perforin-deficient CD4+ T lymphocytes was inhibited only partially by concanamycin A, but was inhibited significantly by antagonistic anti-Fas Ab and anti-Fas ligand Ab. The combination of perforin-deficient effector T lymphocytes and Fas-negative target cells used in the present study provides a novel experimental system for studying the detailed mechanisms of human CTL-mediated cytotoxicity. The present data demonstrate that perforin negative CD4+ CTLs can exert cytotoxicity against Fas-sensitive target cells; however, perforin plays essential roles in Ag-specific cytotoxicity mediated by human CD4+ as well as CD8+ CTLs. PMID- 12574395 TI - Microsatellite instability and suppressed DNA repair enzyme expression in rheumatoid arthritis. AB - Reactive oxygen and nitrogen are produced by rheumatoid arthritis (RA) synovial tissue and can potentially induce mutations in key genes. Normally, this process is prevented by a DNA mismatch repair (MMR) system that maintains sequence fidelity during DNA replication. Key members of the MMR system include MutSalpha (hMSH2 and hMSH6) and MutSbeta (hMSH2 and hMSH3). To provide evidence of DNA damage in inflamed synovium, we analyzed synovial tissues for microsatellite instability (MSI). MSI was examined by PCR on genomic DNA of paired synovial tissue and peripheral blood cells of RA patients using specific primer sequences for five key microsatellites. Surprisingly, abundant MSI was observed in RA synovium compared with osteoarthritis tissue. Western blot analysis for the expression of MMR proteins demonstrated decreased hMSH6 and increased hMSH3 in RA synovium. To evaluate potential mechanisms of MMR regulation in arthritis, fibroblast-like synoviocytes (FLS) were isolated from synovial tissues and incubated with the NO donor S-nitroso-N-acetylpenicillamine. Western blot analysis demonstrated constitutive expression of hMSH2, 3, and 6 in RA and osteoarthritis FLS. When FLS were cultured with S-nitroso-N-acetylpenicillamine, the pattern of MMR expression in RA synovium was reproduced (high hMSH3, low hMSH6). Therefore, oxidative stress can relax the DNA MMR system in RA by suppressing hMSH6. Decreased hMSH6 can subsequently interfere with repair of single base mutations, which is the type observed in RA. We propose that oxidative stress not only creates DNA adducts that are potentially mutagenic, but also suppresses the mechanisms that limit the DNA damage. PMID- 12574396 TI - Perforin-dependent brain-infiltrating cytotoxic CD8+ T lymphocytes mediate experimental cerebral malaria pathogenesis. AB - Experimental cerebral malaria (ECM) resulting from Plasmodium berghei ANKA infection involves T lymphocytes. However, the mechanisms of T cell-mediated pathogenesis remain unknown. We found that, in contrast to ECM-susceptible C57BL6 mice, perforin-deficient (PFP-KO) mice were resistant to ECM in the absence of brain lesions, whereas cytoadherence of parasitized erythrocytes and massive accumulation of activated/effector CD8 lymphocytes were observed in both groups of mice. ECM is induced in PFP-KO mice after adoptive transfer of cytotoxic CD8+ cells from infected C57BL6 mice, which were directed to the brain of PFP-KO mice. This specific recruitment might involve chemokine/chemokine receptors, since their expression was up-regulated on activated CD8 cells, and susceptibility to ECM was delayed in CCR5-KO mice. Thus, lymphocyte cytotoxicity and cell trafficking are key players in ECM pathogenesis. PMID- 12574397 TI - T cell reactivity to MHC class II-bound self peptides in systemic lupus erythematosus-prone MRL/lpr mice. AB - The epitopes recognized by pathogenic T cells in systemic autoimmune disease remain poorly defined. Certain MHC class II-bound self peptides from autoimmune MRL/lpr mice are not found in eluates from class II molecules of MHC-identical C3H mice. Eleven of 16 such peptides elicited lymph node cell and spleen cell T cell proliferation in both MRL/lpr (stimulation index = 2.03-5.01) and C3H mice (stimulation index = 2.03-3.75). IL-2 and IFN-gamma production were detected, but not IL-4. In contrast to what was seen after immunization, four self peptides induced spleen cell proliferation of T cells from naive MRL/lpr, but not from C3H and C57BL/6.H2(k), mice. These peptides were derived from RNA splicing factor SRp20, histone H2A, beta(2)-microglobulin, and MHC class II I-A(k)beta. The first three peptides were isolated from I-E(k) molecules and the last peptide was bound to I-A(k). T cell responses, evident as early as 1 mo of age, depended on MHC class II binding motifs and were inhibited by anti-MHC class II Abs. Thus, although immunization can evoke peripheral self-reactive T cells in normal mice, the presence in MRL/lpr mice of spontaneous T cells reactive to certain MHC-bound self peptides suggests that these T cells actively participate in systemic autoimmunity. Peptides eluted from self MHC class II molecules may yield important clues to T cell epitopes in systemic autoimmunity. PMID- 12574398 TI - In vivo efficacy of anti-glycoprotein 41, but not anti-glycoprotein 120, immunotoxins in a mouse model of HIV infection. AB - Immunotoxins (ITs) targeting the HIV envelope protein are among the most efficacious antiviral therapies when tested in vitro. Yet a first-generation IT targeted to gp120, CD4-PE40 (chimeric immunotoxin using CD4 and the translocation and enzymatic domains of Pseudomonas exotoxin A), showed limited promise in initial clinical testing, highlighting the need for improved ITs. We have used a new mouse model of HIV infection to test the comparative efficacy of anti-HIV ITs targeted to gp120 or to gp41. Irradiated SCID/nonobese diabetic mice are injected with a tumor of human CD4(+) cells susceptible to infection and at a separate site persistently HIV-infected cells. The spread of infection from infected to susceptible tumor is monitored by plasma p24 and the presence of HIV-infected cells in the spleen. Anti-gp41 ITs in combination with tetrameric CD4-human Ig fusion protein have pronounced anti-HIV effects. Little if any anti-HIV efficacy was found with either CD4-PE40 or an Ab-targeted anti-gp120 IT. These data support continued exploration of the utility of ITs for HIV infection, particularly the use of anti-gp41 ITs in combination with soluble CD4 derivatives. PMID- 12574399 TI - Transplantation of olfactory ensheathing cells into spinal cord lesions restores breathing and climbing. AB - One of the most devastating effects of damage to the upper spinal cord is the loss of the ability to breathe; patients suffering these injuries can be kept alive only with assisted ventilation. No known method for repairing these injuries exists. We report here the return of supraspinal control of breathing and major improvements in climbing after the application of a novel endogenous matrix transfer method. This method permits efficient transfer and retention of cultured adult rat olfactory ensheathing cells when transplanted into large lesions that destroy all tracts on one side at the upper cervical level of the adult rat spinal cord. This demonstrates that transplantation can produce simultaneous repair of two independent spinal functions. PMID- 12574400 TI - Generation of functional inhibitory neurons in the adult rat hippocampus. AB - Several thousand new neurons are produced each day in the adult mammalian hippocampus, among which only excitatory granule cells (GCs) have thus far been identified. In the present study, we used mutant Semliki Forest Virus vectors to express enhanced green fluorescent protein in the hippocampus, and observed that approximately 14% of newly generated neurons in the dentate gyrus of adult rats are GABAergic basket cells (BCs). With the use of double whole-cell patch-clamp recordings from BC-GC pairs in hippocampal slices, we demonstrate that newly generated BCs in the dentate gyrus form inhibitory synapses with principal GCs. These data show for the first time that functional inhibitory neurons are recruited in the dentate gyrus of adult rats. PMID- 12574401 TI - Inhibition of mRNA and protein synthesis in the CA1 region of the dorsal hippocampus blocks reinstallment of an extinguished conditioned fear response. AB - Memories are extinguished by the repeated presentation of a conditioned stimulus in the absence of an unconditioned stimulus to which it has been associated. It is believed that extinction establishes a new hierarchy of responses rather than an actual forgetting of the original response, which can usually reappear spontaneously after interruption of the extinction process. In this study, our aim was to analyze how profound extinction can be. Rats were trained in a one trial, step-down inhibitory avoidance paradigm and then were exposed to several extinction sessions in which they were allowed to freely explore the apparatus for 30 sec after having stepped down. Extinction was complete enough so that there was no spontaneous recovery, and test session performance could not be enhanced by pharmacological agents with well known facilitative actions on retrieval. After being submitted to a new training session, control animals reacquired the avoidance response; however, animals failed to do so after receiving bilateral intra-CA1 infusions of either the protein synthesis inhibitor anisomycin or the mRNA synthesis blocker 5,6-dichloro-1-beta-d-ribofuranosyl benzimidazole 15 min before the retraining session. Our results indicate that extinction can be carried to a point at which reinstallment of the conditioned response requires, like the original learning, de novo gene expression and protein synthesis in the CA1 region of the dorsal hippocampus. PMID- 12574402 TI - Time-dependent increases in brain-derived neurotrophic factor protein levels within the mesolimbic dopamine system after withdrawal from cocaine: implications for incubation of cocaine craving. AB - Using a rat model of drug craving, we found that the responsiveness to cocaine cues progressively increases or incubates over the first 60 d of cocaine withdrawal. Here we studied whether alterations in brain-derived neurotrophic factor (BDNF) protein levels within the mesolimbic dopamine system are associated with this incubation phenomenon. BDNF is involved in synaptic plasticity and was found to enhance responding for cues associated with natural rewards. Rats were trained to press a lever to receive intravenous cocaine or oral sucrose for 6 hr/d for 10 d; each earned reward was paired with a tone-light cue. Resumption of lever-pressing behavior was then assessed on days 1, 30, or 90 of reward withdrawal. First, resistance to extinction was assessed during 6 hr in which lever presses were not reinforced and the cue was absent. Second, cue-induced reinstatement was assessed after extinction during 1 hr in which responding led to cue presentations. Other rats were killed without testing on days 1, 30, and 90 of reward withdrawal, and BDNF and nerve growth factor (NGF) protein levels were measured in the ventral tegmental area (VTA), accumbens, and amygdala. Lever pressing during extinction and cue-induced reinstatement tests of cocaine craving progressively increased after cocaine withdrawal. Time-dependent changes also were observed during the tests for sucrose craving, with maximal responding on day 30. BDNF, but not NGF, levels in the VTA, accumbens, and amygdala progressively increased after cocaine, but not sucrose, withdrawal. Time dependent increases in BDNF levels may lead to synaptic modifications that underlie enhanced responsiveness to cocaine cues after prolonged withdrawal periods. PMID- 12574403 TI - The RAS effector RIN1 modulates the formation of aversive memories. AB - RAS proteins are critical regulators of mitosis and are mutationally activated in many human tumors. RAS signaling is also known to mediate long-term potentiation (LTP) and long-term memory formation in postmitotic neurons, in part through activation of the RAF-MEK-ERK pathway. The RAS effector RIN1 appears to function through competitive inhibition of RAS-RAF binding and also through diversion of RAS signaling to alternate pathways. We show that RIN1 is preferentially expressed in postnatal forebrain neurons in which it is localized in dendrites and physically associated with RAS, suggesting a role in RAS-mediated postsynaptic neuronal plasticity. Mice with an Rin1 gene disruption showed a striking enhancement in amygdala LTP. In addition, two independent behavioral tests demonstrated elevated amygdala-dependent aversive memory in Rin1(-/-) mice. These results indicate that RIN1 serves as an inhibitory modulator of neuronal plasticity in aversive memory formation. PMID- 12574404 TI - Ca2+ signaling in mouse cortical neurons studied by two-photon imaging and photoreleased inositol triphosphate. AB - IP(3)-mediated Ca(2+) release is a crucial neuronal signaling mechanism that has not been extensively characterized in the mammalian cerebral cortex. We used two photon, video-rate microscopy to image Ca(2+) signals evoked by photoreleased IP(3) in pyramidal neurons of mouse prefrontal cortex. Ca(2+) responses to photoreleased IP(3) varied greatly between different neurons; however, within IP(3)-responsive neurons, the soma invariably showed highest sensitivity, with signals increasing nonlinearly with [IP(3)]. Responses to paired photorelease displayed inhibition, whereas IP(3)-evoked Ca(2+) liberation was potentiated by Ca(2+) entry during action potentials and vice versa. IP(3)-mediated Ca(2+) signals strongly inhibited spike firing through activation of K(+) membrane conductance. Metabotropic signaling via the phosphoinositide pathway thus serves as a powerful and sustained modulator of excitability in cortical neurons and displays complex reciprocal interactions between electrical and chemical signals. PMID- 12574405 TI - Accelerated hippocampal spreading depression and enhanced locomotory activity in mice with astrocyte-directed inactivation of connexin43. AB - Using a human glial fibrillary acidic protein (hGFAP) promoter-driven cre transgene, we have achieved efficient inactivation of a floxed connexin43 (Cx43) gene in astrocytes of adult mice. The loss of Cx43 expression was monitored in a cell-autonomous manner via conditional replacement of the Cx43-coding region by a lacZ reporter gene. In this way, we bypassed the early postnatal lethality previously reported for Cx43 null mice and characterized the phenotypic consequences of Cx43 deficiency in the CNS. Mice lacking Cx43 in astrocytes were viable and showed no evidence of either neurodegeneration or astrogliosis. Spreading depression (SD) is a pathophysiological phenomenon observed in the CNS that is characterized by a propagating wave of depolarization followed by neuronal inactivation. Inhibitors of gap junctional communication have previously been shown to block initiation and propagation of SD. In contrast, we observed an increase in the velocity of hippocampal SD in the stratum radiatum of mice lacking Cx43 in astrocytes. In the same brain subregion, dye-coupling experiments revealed a reduction in overall astrocytic intercellular communication by approximately 50%. This strongly suggests separate and different neuronal and glial contributions of gap junctional intercellular communication to SD. Concomitant with increased velocity of spreading depression, we observed enhanced locomotory activity in mice lacking Cx43 in astrocytes. PMID- 12574406 TI - An extranuclear locus of cAMP-dependent protein kinase action is necessary and sufficient for promotion of spiral ganglion neuronal survival by cAMP. AB - We showed previously that cAMP is a survival-promoting stimulus for cultured postnatal rat spiral ganglion neurons (SGNs) and that depolarization promotes SGN survival in part via recruitment of cAMP signaling. We here investigate the subcellular locus of cAMP prosurvival signaling. Transfection of GPKI, a green fluorescent protein (GFP)-tagged cAMP-dependent protein kinase (PKA) inhibitor, inhibits the ability of the permeant cAMP analog cpt-cAMP [8-(4-chlorophenylthio) cAMP] to promote survival, indicating that PKA activity is necessary. Transfection of GFP-tagged PKA (GPKA) is sufficient to promote SGN survival, but restriction of GPKA to the nucleus by addition of a nuclear localization signal (GPKAnls) almost completely abrogates its prosurvival effect. In contrast, GPKA targeted to the extranuclear cytoplasm by addition of a nuclear export signal (GPKAnes) promotes SGN survival as effectively as does GPKA. Moreover, GPKI targeted to the nucleus lacks inhibitory effect on SGN survival attributable to cpt-cAMP or depolarization. These data indicate an extranuclear target of PKA for promotion of neuronal survival. Consistent with this, we find that dominant inhibitory CREB mutants inhibit the prosurvival effect of depolarization but not that of cpt-cAMP. SGN survival is compromised by overexpression of the proapoptotic regulator Bad, previously shown to be phosphorylated in the cytoplasm by PKA. This Bad-induced apoptosis is prevented by cpt-cAMP or by cotransfection of GPKA or of GPKAnes but not of GPKAnls. Thus, cAMP prevents SGN death through a cytoplasmic as opposed to nuclear action, and inactivation of Bad proapoptotic function is a mechanism by which PKA can prevent neuronal death. PMID- 12574407 TI - Oxytocin regulates neurosteroid modulation of GABA(A) receptors in supraoptic nucleus around parturition. AB - In this study, we investigate how neurosteroid sensitivity of GABA(A) receptors (GABA(A)Rs) is regulated. We examined this issue in neurons of the supraoptic nucleus (SON) of the rat and found that, during parturition, the GABA(A)Rs become insensitive to the neurosteroid allopregnanolone attributable to a shift in the balance between the activities of endogenous Ser/Thr phosphatase and PKC. In particular, a constitutive endogenous tone of oxytocin within the SON after parturition suppressed neurosteroid sensitivity of GABA(A)Rs via activation of PKC. Vice versa before parturition, during late pregnancy, application of exogenous oxytocin brings the GABA(A)Rs from a neurosteroid-sensitive mode toward a condition in which the receptors are not sensitive. This indicates that there may be an inverse causal relationship between the extent to which the GABA(A)R or one of its interacting proteins is phosphorylated and the neurosteroid sensitivity of the GABA(A)R. Neurosteroid sensitivity was not affected by changes in subunit composition of GABA(A)Rs known to occur concurrently in these cells. PMID- 12574408 TI - Surface expression of GluR-D AMPA receptor is dependent on an interaction between its C-terminal domain and a 4.1 protein. AB - Dynamic regulation of the number and activity of AMPA receptors is believed to underlie many forms of synaptic plasticity and is presumably mediated by specific protein-protein interactions involving the C-terminal domain of the receptor. Several proteins interacting with the C-terminal tails of the glutamate receptor (GluR)-A and GluR-B subunits have been identified and implicated in the regulation of endocytosis and exocytosis, clustering, and anchoring of AMPA receptors to the cytoskeleton. In contrast, little is known of the molecular interactions of the GluR-D subunit, or of the mechanisms regulating the traffic of GluR-D-containing AMPA receptors. We analyzed the subcellular localization of homomeric GluR-D receptors carrying C-terminal deletions in transfected human embryonic kidney (HEK) 293 cells and in primary neurons by immunofluorescence microscopy and ELISA. A minimal requirement for a 14-residue cytoplasmic segment for the surface expression of homomeric GluR-D receptors was identified. Previously, a similar region in the GluR-A subunit was implicated in an interaction with 4.1 family proteins. Coimmunoprecipitation demonstrated that GluR-D associated with 4.1 protein(s) in both HEK293 cells and rat brain. Moreover, glutathione S-transferase pull-down experiments showed that the same 14 residue segment is critical for 4.1 binding to GluR-A and GluR-D. Point mutations within this segment dramatically decreased the surface expression of GluR-D in HEK293 cells, with a concomitant loss of the 4.1 interaction. Our findings demonstrate a novel molecular interaction for the GluR-D subunit and suggest that the association with the 4.1 family protein(s) plays an essential role in the transport to and stabilization of GluR-D-containing AMPA receptors at the cell surface. PMID- 12574409 TI - Stimulation of the calcitonin gene-related peptide enhancer by mitogen-activated protein kinases and repression by an antimigraine drug in trigeminal ganglia neurons. AB - Calcitonin gene-related peptide (CGRP) is involved in the underlying pathophysiology of all vascular headaches, including migraines. Elevated levels of CGRP during migraine are restored to normal coincident with headache relief after treatment with the antimigraine drug sumatriptan. We have used primary cultures of trigeminal neurons under conditions simulating migraine pathology and therapy to study the mechanisms controlling the CGRP promoter. Using reporter genes in transient transfection assays, we demonstrate that an 18 bp enhancer containing a helix-loop-helix element is both necessary and sufficient for full promoter activity. NGF treatment and cotransfection with an upstream activator of the extracellular signal-regulated MAP kinases (MAPKs) activated the enhancer. Treatment with sumatriptan repressed NGF- and MAPK-stimulated CGRP promoter activity. Repression was also observed using a synthetic MAPK-responsive reporter gene. Sumatriptan regulation of CGRP gene expression did not couple to a G(i)/G(o) pathway, but rather caused a prolonged increase in intracellular calcium. The importance of the prolonged calcium signal in repression of MAPK activity was demonstrated by using the ionophore ionomycin to mimic sumatriptan action. We propose that activation of MAPK pathways may increase CGRP gene expression during migraine, and that sumatriptan can diametrically oppose that activation via a prolonged elevation of intracellular calcium. PMID- 12574410 TI - D5 (not D1) dopamine receptors potentiate burst-firing in neurons of the subthalamic nucleus by modulating an L-type calcium conductance. AB - Dopamine is a crucial factor in basal ganglia functioning. In current models of basal ganglia, dopamine is postulated to act on striatal neurons. However, it may also act on the subthalamic nucleus (STN), a key nucleus in the basal ganglia circuit. The data presented here were obtained in brain slices using whole-cell patch clamp. They reveal that D5 dopamine receptors strengthen electrical activity in the subset of subthalamic neurons endowed with burst-firing capacity, resulting in longer discharges of spontaneous or evoked bursts. To distinguish between D1 and D5 subtypes, the action of agonists in the D1/D5 receptor family was first investigated on rat subthalamic neurons. Single-cell reverse transcription-PCR profiling showed that burst-competent neurons only expressed D5 receptors. Accordingly, receptors localized in postsynaptic membranes within the STN were labeled by a D5-specific antibody. Second, agonists in the D1/D5 family were tested in mouse brain slices. It was found that these agonists were active in D1 receptor knock-out mice in a similar way to wild-type mice or rats. This proved that D5 rather than D1 receptors were involved. Pharmacological tools (dihydropyridines, omega-conotoxins, and calciseptine) were used to identify the target of D5 receptors as an L-type channel. This was reached via G-protein and protein kinase A. The action of dopamine on D5 receptors therefore shapes neuronal activity. It contributes to normal information processing in basal ganglia outside striatum. This finding may be useful in drug therapy for various disorders involving changes in STN activity, such as Parkinson's disease and related disorders. PMID- 12574411 TI - Interaction of calcineurin and type-A GABA receptor gamma 2 subunits produces long-term depression at CA1 inhibitory synapses. AB - Long-term depression (LTD) is an activity-dependent weakening of synaptic efficacy at individual inhibitory synapses, a possible cellular model of learning and memory. Here, we show that the induction of LTD of inhibitory transmission recruits activated calcineurin (CaN) to dephosphorylate type-A GABA receptor (GABA(A)Rs) via the direct binding of CaN catalytic domain to the second intracellular domain of the GABA(A)R-gamma(2) subunits. Prevention of the CaN GABA(A) receptor complex formation by expression of an autoinhibitory domain of CaN in the hippocampus of transgenic mice blocks the induction of LTD. Conversely, genetic expression of the CaN catalytic domain in the hippocampus depresses inhibitory synaptic responses, occluding LTD. Thus, an activity dependent physical and functional interaction between CaN and GABA(A) receptors is both necessary and sufficient for inducing LTD at CA1 individual inhibitory synapses. PMID- 12574412 TI - Involvement of actin polymerization in vesicle recruitment at the calyx of Held synapse. AB - Depletion and replenishment of pools of synaptic vesicles are important determinants of short-term synaptic plasticity, but the underlying molecular mechanisms are not yet clear. As a first step toward understanding the process of vesicle recruitment, we have applied various specific agents directly to the presynaptic terminal of the calyx of Held synapse. Here we show that the nonhydrolyzable ATP analog ATP-gammaS retards the recovery from vesicle pool depletion, as does latrunculin A. Phalloidin has no effects on recovery, suggesting that dynamic actin reorganization is not necessary. Unexpectedly, neither N-ethylmaleimide nor staurosporine affected the recovery, calling into question the role of N-ethylmaleimide-sensitive factor and protein kinases. The results suggest that intact actin polymerization is involved in vesicle recruitment. PMID- 12574413 TI - How AMPA receptor desensitization depends on receptor occupancy. AB - AMPA-type glutamate receptors mediate fast excitatory transmission at many central synapses, and rapid desensitization of these receptors can shape the decay of synaptic currents and limit the fidelity of high-frequency synaptic transmission. Here we use a combination of fast glutamate application protocols and kinetic simulations to determine how AMPA receptor desensitization depends on the number of subunits occupied by glutamate. We show that occupancy of a single subunit is sufficient to desensitize AMPA-type channels and that receptors with one to four glutamates bound enter desensitization at similar rates. We find that recovery from desensitization follows a similar sigmoid time course for channels with two to four glutamates bound but is faster and exponential for singly occupied channels. The results suggest that desensitization, at intermediate and high glutamate concentrations, is accompanied by two conformational changes that slow glutamate dissociation. We propose a kinetic scheme that accurately predicts several types of experimental results and differs significantly from previous models in the assignment of affinities for binding to closed and desensitized states. We conclude that desensitization involves a rearrangement that stabilizes the binding domains of one subunit in each dimer in a partially closed conformation. This stabilization likely results from an interaction at the dimer dimer interface between the binding domains of adjacent subunits. PMID- 12574414 TI - Functional autaptic neurotransmission in fast-spiking interneurons: a novel form of feedback inhibition in the neocortex. AB - Autapses are synapses made by a neuron onto itself. Although morphological evidence for existence of autapses has been reported in several brain areas, it is not known whether such self-innervation in the neocortex is functional and robust. Here we report that GABAergic autaptic activity is present in fast spiking, but not in low-threshold spiking, interneurons of layer V in neocortical slices. Recordings made with the perforated-patch technique, in which physiological intracellular chloride homeostasis was unperturbed, demonstrated that autaptic activity has significant inhibitory effects on repetitive firing and increased the current threshold for evoking action potentials. These results show that autapses are not rudimentary nonfunctional structures, but rather they provide a novel and powerful form of feedback inhibitory synaptic transmission in one class of cortical interneurons. PMID- 12574415 TI - Dopamine enhances EPSCs in layer II-III pyramidal neurons in rat prefrontal cortex. AB - Dopaminergic inputs to the prefrontal cortex (PFC) are important for the integration of neuronal signals, the formation of working memory, and the establishment of memory fields. A detailed characterization of cellular mechanisms underlying the effects of dopamine on PFC is still emerging. We have examined how dopamine affects excitatory synaptic transmission in the PFC using whole-cell patch-clamp recording from visually identified layer II-III pyramidal cells in vitro. Bath application of dopamine significantly enhanced EPSC amplitudes. Pharmacologically isolated AMPA and NMDA receptor-mediated EPSCs were increased to a similar extent. Application of the specific D1-like receptor agonist SKF38393 [(+/-)-1-phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol hydrobromide] significantly increased EPSC amplitude, whereas the D2-like receptor agonist quinpirole had no effect. Responses to pressure-applied glutamate were also enhanced by dopamine, indicating a postsynaptic mechanism. Inclusion of the Ca(2+) chelator BAPTA in the recording pipette blocked the dopamine enhancement. When the PKA inhibitory peptide PKI [5-24] was included in the recording pipette, dopamine did not affect EPSCs. Similarly, when the Ca(2+)/calmodulin-kinase II (CaMKII) inhibitory peptide was present in the pipette, dopamine enhancement of EPSCs was not observed in any of the cells tested. These results indicate that EPSC enhancement may be attributable to a postsynaptic signaling cascade involving Ca(2+), PKA, and CaMKII. PMID- 12574416 TI - Early exposure to common anesthetic agents causes widespread neurodegeneration in the developing rat brain and persistent learning deficits. AB - Recently it was demonstrated that exposure of the developing brain during the period of synaptogenesis to drugs that block NMDA glutamate receptors or drugs that potentiate GABA(A) receptors can trigger widespread apoptotic neurodegeneration. All currently used general anesthetic agents have either NMDA receptor-blocking or GABA(A) receptor-enhancing properties. To induce or maintain a surgical plane of anesthesia, it is common practice in pediatric or obstetrical medicine to use agents from these two classes in combination. Therefore, the question arises whether this practice entails significant risk of inducing apoptotic neurodegeneration in the developing human brain. To begin to address this problem, we have administered to 7-d-old infant rats a combination of drugs commonly used in pediatric anesthesia (midazolam, nitrous oxide, and isoflurane) in doses sufficient to maintain a surgical plane of anesthesia for 6 hr, and have observed that this causes widespread apoptotic neurodegeneration in the developing brain, deficits in hippocampal synaptic function, and persistent memory/learning impairments. PMID- 12574417 TI - Fibroblast growth factor receptor 3 signaling regulates the onset of oligodendrocyte terminal differentiation. AB - Fibroblast growth factor receptor (FGFR) signaling is essential for nervous system development. We have shown that, in the normal postnatal brain, the spatial and temporal expression pattern of FGFR3 parallels the appearance of differentiated oligodendrocytes and that in culture FGFR3 is expressed maximally at the critical stage in the lineage at which oligodendrocyte late progenitors (Pro-OLs) enter terminal differentiation. Therefore, FGFR3 expression is positioned ideally to have an impact on oligodendrocyte differentiation. In support of this we show that, during the onset and active phase of myelination in FGFR3-deficient mice, there are reduced numbers of differentiated oligodendrocytes in the forebrain, cerebellum, hindbrain, and spinal cord. Furthermore, myelination is delayed in parallel. Delay of oligodendrocyte differentiation also is observed in primary cell culture from this mutant. On the other hand, no differences are observed in the survival or proliferation of oligodendrocyte progenitors. This suggests that the decrease in the number of differentiated oligodendrocytes is attributable to a delay in the timing of their differentiation process. Astrocytes also express FGFR3, and in mice lacking FGFR3 there is an enhancement of the astrocytic marker glial fibrillary acidic protein expression in a region-specific manner. Thus our findings suggest that there are cell type- and region-specific functions for FGFR3 signaling and in particular emphasize a prominent role for FGFR3 as part of a system regulating the onset of oligodendrocyte terminal differentiation. PMID- 12574418 TI - Locally born olfactory bulb stem cells proliferate in response to insulin-related factors and require endogenous insulin-like growth factor-I for differentiation into neurons and glia. AB - After late embryogenesis, new neurons are continuously added to the olfactory bulb (OB) from stem cells located in the forebrain subventricular zone. Nonetheless, stem cells have not been described within the embryonic olfactory bulb. Here we report the isolation of local olfactory bulb stem cells from the embryonic day 12.5-14.5 mouse embryo. These cells were 99.2% nestin positive and proliferated extensively in culture to at least 150 cell doublings. Clonal analysis demonstrated that neurons (TuJ1(+)), astrocytes (GFAP(+)), and oligodendrocytes (O4(+)) could be generated from single-plated cells, indicating that they are multipotent. At least 90% of proliferating cells expressed insulin like growth factor-I (IGF-I), (pro)insulin, and their cognate receptors; these growth factors collaborated with fibroblast growth factor-2 plus epidermal growth factor (EGF) to promote stem cell proliferation and sphere formation. Cells from Igf-I(-)/- mice, however, proliferated as extensively as did Igf-I(+/+) cells. Differentiation and survival of stem cell-generated neurons and glia showed strong dependence on exogenous IGF-I, but oligodendrocyte differentiation also required insulin at low concentration. Furthermore, the percentages of stem cell generated neurons, astrocytes, and oligodendrocytes were markedly lower in the cultures prepared from the Igf-I(-)/- mice compared with those of Igf-I(+/+). Concordantly, lack of IGF-I resulted in abnormal formation of the olfactory bulb mitral cell layer and altered radial glia morphology. These results support the presence within the embryonic mouse olfactory bulb of stem cells with specific requirements for insulin-related growth factors for proliferation or differentiation. They demonstrate that IGF-I is an endogenous factor regulating the differentiation of stem and other precursor cells within the olfactory bulb. PMID- 12574419 TI - Role of the G-protein-coupled receptor GPR12 as high-affinity receptor for sphingosylphosphorylcholine and its expression and function in brain development. AB - Lysophospholipids are bioactive molecules influencing numerous cellular processes such as proliferation, differentiation, and motility. As extracellular ligands, they interact with specific members of the G-protein-coupled receptor family. We show in this paper that the lysophospholipid sphingosylphosphorylcholine is a high-affinity ligand for the orphan G-protein-coupled receptor GPR12. Heterologous expression of GPR12 in Chinese hamster ovary cells and in frog oocytes revealed a high-affinity interaction with sphingosylphosphorylcholine in the nanomolar range. Blockade of its action by pertussis toxin was taken as evidence that GPR12 is coupled to an inhibitory G-protein. In the adult mouse brain, GPR12 was expressed in the limbic system. During mouse embryonal development, GPR12 transcripts were detected in the CNS, especially in areas where neuronal differentiation occurs. Consistent with this we found that cultures of embryonal cerebral cortical neurons responded to sphingosylphosphorylcholine with an increase in synaptic contacts. The GPR12 expressing hippocampal cell line HT22 reacted to sphingosylphophorylcholine with an increase in cell proliferation and cell clustering. Other receptors known to interact at nanomolar concentrations with sphingosylphosphorycholine were expressed neither in the developing cerebral cortex nor in the HT22 cell line. We therefore hypothesize that sphingosylphosphorylcholine, most likely by interaction with GPR12, has positive effects on the differentiation and maturation of postmitotic neurons and that it may also influence the proliferation of neuronal precursor cells. PMID- 12574420 TI - Neuron-to-glia signaling mediated by excitatory amino acid receptors regulates ErbB receptor function in astroglial cells of the neuroendocrine brain. AB - Hypothalamic astroglial erbB tyrosine kinase receptors are required for the timely initiation of mammalian puberty. Ligand-dependent activation of these receptors sets in motion a glia-to-neuron signaling pathway that prompts the secretion of luteinizing hormone-releasing hormone (LHRH), the neuropeptide controlling sexual development, from hypothalamic neuroendocrine neurons. The neuronal systems that may regulate this growth factor-mediated back signaling to neuroendocrine neurons have not been identified. Here we demonstrate that hypothalamic astrocytes contain metabotropic receptors of the metabotropic glutamate receptor 5 subtype and the AMPA receptor subunits glutamate receptor 2 (GluR2) and GluR3. As in excitatory synapses, these receptors are in physical association with their respective interacting/clustering proteins Homer and PICK1. In addition, they are associated with erbB-1 and erbB-4 receptors. Concomitant activation of astroglial metabotropic and AMPA receptors results in the recruitment of erbB tyrosine kinase receptors and their respective ligands to the glial cell membrane, transactivation of erbB receptors via a mechanism requiring metalloproteinase activity, and increased erbB receptor gene expression. By facilitating erbB-dependent signaling and promoting erbB receptor gene expression in astrocytes, a neuron-to-glia glutamatergic pathway may represent a basic cell-cell communication mechanism used by the neuroendocrine brain to coordinate the facilitatory transsynaptic and astroglial input to LHRH neurons during sexual development. PMID- 12574421 TI - Spontaneous calcium transients in developing cortical neurons regulate axon outgrowth. AB - Growth cones of cortical axons pause for many hours in preparation for axon branching. They become large and complex compared with small advancing growth cones. We wanted to investigate whether calcium transients regulate the advance of mammalian CNS growth cones. We found that spontaneous calcium transients in developing cortical neurons have characteristic patterns, frequencies, and amplitudes. Importantly, neurons with large paused growth cones exhibit high frequency spontaneous calcium transients, which are rare in those with small advancing growth cones. The incidence, frequencies, and amplitudes of calcium transients are inversely related to rates of axon outgrowth. The transients are mediated primarily by L-type voltage-gated calcium channels, and silencing them with channel blockers promotes axon outgrowth. Thus calcium transients regulate growth cone advance by direct effects on the growth cone. PMID- 12574422 TI - Nonrenewal of neurons in the cerebral neocortex of adult macaque monkeys. AB - The concept that, after developmental periods, neocortical neurons become numerically stable and are normally nonrenewable has been challenged by a report of continuous neurogenesis in the association areas of the cerebral cortex in the adult Macaque monkey. Therefore, we have reexamined this issue in two different Macaque species using the thymidine analog bromodeoxyuridine (BrdU) as an indicator of DNA replication during cell division. We found several BrdU+/NeuN+ (neuronal nuclei) double-labeled cells, but cortical neurons, distinguished readily by their size and cytological and immunohistochemical properties, were not BrdU positive. We examined in detail the frontal cortex, where it is claimed that the largest daily addition of neurons has been made, but did not see migratory streams or any sign of addition of new neurons. Thus, we concluded that, in the normal condition, cortical neurons of adult primates, similar to other mammalian species, are neither supplemented nor renewable. PMID- 12574423 TI - The functional consequences of changes in the strength and duration of synaptic inputs to oscillatory neurons. AB - We studied the effect of synaptic inputs of different amplitude and duration on neural oscillators by simulating synaptic conductance pulses in a bursting conductance-based pacemaker model and by injecting artificial synaptic conductance pulses into pyloric pacemaker neurons of the lobster stomatogastric ganglion using the dynamic clamp. In the model and the biological neuron, the change in burst period caused by inhibitory and excitatory inputs of increasing strength saturated, such that synaptic inputs above a certain strength all had the same effect on the firing pattern of the oscillatory neuron. In contrast, increasing the duration of the synaptic conductance pulses always led to changes in the burst period, indicating that neural oscillators are sensitive to changes in the duration of synaptic input but are not sensitive to changes in the strength of synaptic inputs above a certain conductance. This saturation of the response to progressively stronger synaptic inputs occurs not only in bursting neurons but also in tonically spiking neurons. We identified inward currents at hyperpolarized potentials as the cause of the saturation in the model neuron. Our findings imply that activity-dependent or modulator-induced changes in synaptic strength are not necessarily accompanied by changes in the functional impact of a synapse on the timing of postsynaptic spikes or bursts. PMID- 12574424 TI - Formation of complement membrane attack complex in mammalian cerebral cortex evokes seizures and neurodegeneration. AB - The complement system consists of >30 proteins that interact in a carefully regulated manner to destroy invading bacteria and prevent the deposition of immune complexes in normal tissue. This complex system can be activated by diverse mechanisms proceeding through distinct pathways, yet all converge on a final common pathway in which five proteins assemble into a multimolecular complex, the membrane attack complex (MAC). The MAC inserts into cell membranes to form a functional pore, resulting in ion flux and ultimately osmotic lysis. Immunohistochemical evidence of the MAC decorating neurons in cortical gray matter has been identified in multiple CNS diseases, yet the deleterious consequences, if any, of MAC deposition in the cortex of mammalian brain in vivo are unknown. Here we demonstrate that the sequential infusion of individual proteins of the membrane attack pathway (C5b6, C7, C8, and C9) into the hippocampus of awake, freely moving rats induced both behavioral and electrographic seizures as well as cytotoxicity. The onset of seizures occurred during or shortly after the infusion of C8/C9. Neither seizures nor cytotoxicity resulted from the simultaneous infusion of all five proteins premixed in vitro. The requirement for the sequential infusion of all five proteins together with the temporal relationship of seizure onset to infusions of C8/C9 implies that the MAC was formed in vivo and triggered both seizures and cytotoxicity. Deposition of the complement MAC in cortical gray matter may contribute to epileptic seizures and cell death in diverse diseases of the human brain. PMID- 12574425 TI - Transient and progressive electrophysiological alterations in the corticostriatal pathway in a mouse model of Huntington's disease. AB - Alterations in the corticostriatal pathway may precede symptomatology and striatal cell death in Huntington's disease (HD) patients. Here we examined spontaneous EPSCs in striatal medium-sized spiny neurons in slices from a mouse model of HD (R6/2). Spontaneous EPSC frequency was similar in young (3-4 weeks) transgenics and controls but decreased significantly in transgenics when overt behavioral symptoms began (5-7 weeks) and was most pronounced in severely impaired transgenics (11-15 weeks). These differences were maintained after bicuculline or tetrodotoxin, indicating they were specific to glutamatergic input and likely presynaptic in origin. Decreases in presynaptic and postsynaptic protein markers, synaptophysin and postsynaptic density-95, occurred in 11-15 week R6/2 mice, supporting the electrophysiological results. Furthermore, isolated, large-amplitude synaptic events (>100 pA) occurred more frequently in transgenic animals, particularly at 5-7 weeks, suggesting additional dysregulation of cortical inputs. Large events were blocked by tetrodotoxin, indicating a possible cortical origin. Addition of bicuculline and 4 aminopyridine facilitated the occurrence of large events. Riluzole, a compound that decreases glutamate release, reduced these events. Together, these observations indicate that both progressive and transient alterations occur along the corticostriatal pathway in experimental HD. These alterations are likely to contribute to the selective vulnerability of striatal medium-sized spiny neurons. PMID- 12574426 TI - Circuitry underlying regulation of the serotonergic system by swim stress. AB - The dorsal raphe nucleus (DR)-serotonin (5-HT) system has been implicated in depression and is dramatically affected by swim stress, an animal model with predictive value for antidepressants. Accumulating evidence implicates the stress related neuropeptide corticotropin-releasing factor (CRF) in the effect of swim stress on this system. This study investigated neural circuits within the DR that are activated by swim stress as revealed by neuronal expression of the immediate early gene, c-fos. Swim stress increased c-fos expression in the dorsolateral subregion of the DR. The majority of c-fos-expressing neurons were doubly labeled for GABA (85 +/- 5%), whereas relatively few were immunolabeled for 5-HT (4 +/- 1%), glutamate (0.5 +/- 0.3%) or calbindin (1.5 +/- 0.3%). Dual immunohistochemical labeling revealed that c-fos-expressing neurons in the dorsolateral DR were enveloped by dense clusters of CRF-immunoreactive fibers and also contained immunolabeling for CRF receptor, suggesting that c-fos-expressing neurons in the DR were specifically targeted by CRF. Consistent with this, the CRF receptor 1 antagonist, antalarmin, prevented swim-stress-elicited c-fos expression in the dorsolateral DR. Together with previous findings that both swim stress and CRF decrease 5-HT release in certain forebrain regions, these results suggest that swim stress engages CRF inputs to GABA neurons in the dorsolateral DR that function to inhibit 5-HT neurons and 5-HT release in the forebrain. This circuitry may underlie some of the acute behavioral responses to swim stress as well as the neuronal plasticity involved in long-term behavioral changes produced by this stress. PMID- 12574427 TI - Suppression of central taste transmission by oral capsaicin. AB - Because intraoral capsaicin is reported to reduce the perceived intensity of certain taste qualities, we investigated whether it affects the central processing of gustatory information. The responses of gustatory neurons in the nucleus tractus solitarius (NTS) to tastant stimuli were recorded before and after lingual application of capsaicin in anesthetized rats. Thirty-four NTS units were characterized as responding best to sucrose (0.3 m), NaCl (0.1 m), citric acid (0.03 m), monosodium glutamate (0.2 m), or quinine (0.001 m). During lingual application of 330 microm capsaicin for 7 min, the firing rate increased for five units and decreased for four units; the remainder were unaffected. Immediately after capsaicin, responses to each tastant were in nearly all cases depressed (mean, 61.5% of control), followed by recovery in most cases. NTS tastant-evoked unit responses were unaffected by lingual application of vehicle (5% ethanol). Capsaicin elicited an equivalent reduction (to 64.5%) in tastant evoked responses of nine additional NTS units recorded in rats with bilateral trigeminal ganglionectomy, arguing against a trigeminally mediated central effect. Furthermore, capsaicin elicited a puncate pattern of plasma extravasation in the tongue that matched the distribution of fungiform papillae. These results support a peripheral site of capsaicin suppression of taste possibly via direct or indirect effects on taste transduction or taste receptor cell excitability. The depressant effect of capsaicin on gustatory transmission might underlie its ability to reduce the perceived intensity of some taste qualities. PMID- 12574428 TI - Evidence from functional neuroimaging of a compensatory prefrontal network in Alzheimer's disease. AB - Previous experiments have found that individuals with Alzheimer's disease (AD) show increased activity in prefrontal regions compared with healthy age-matched controls during cognitive tasks. This has been interpreted as compensatory reallocation of cognitive resources, but direct evidence for a facilitating effect on performance has been lacking. To address this we measured neural activity during semantic and episodic memory tasks in mildly demented AD patients and healthy elderly controls. Controls recruited a left hemisphere network of regions, including prefrontal and temporal cortices in both the semantic and episodic tasks. Patients engaged a unique network involving bilateral dorsolateral prefrontal and posterior cortices. Critically, activity in this network of regions was correlated with better performance on both the semantic and episodic tasks in the patients. This provides the most direct evidence to date that AD patients can use additional neural resources in prefrontal cortex, presumably those mediating executive functions, to compensate for losses attributable to the degenerative process of the disease. PMID- 12574429 TI - Dynamics of gray matter loss in Alzheimer's disease. AB - We detected and mapped a dynamically spreading wave of gray matter loss in the brains of patients with Alzheimer's disease (AD). The loss pattern was visualized in four dimensions as it spread over time from temporal and limbic cortices into frontal and occipital brain regions, sparing sensorimotor cortices. The shifting deficits were asymmetric (left hemisphere > right hemisphere) and correlated with progressively declining cognitive status (p < 0.0006). Novel brain mapping methods allowed us to visualize dynamic patterns of atrophy in 52 high-resolution magnetic resonance image scans of 12 patients with AD (age 68.4 +/- 1.9 years) and 14 elderly matched controls (age 71.4 +/- 0.9 years) scanned longitudinally (two scans; interscan interval 2.1 +/- 0.4 years). A cortical pattern matching technique encoded changes in brain shape and tissue distribution across subjects and time. Cortical atrophy occurred in a well defined sequence as the disease progressed, mirroring the sequence of neurofibrillary tangle accumulation observed in cross sections at autopsy. Advancing deficits were visualized as dynamic maps that change over time. Frontal regions, spared early in the disease, showed pervasive deficits later (>15% loss). The maps distinguished different phases of AD and differentiated AD from normal aging. Local gray matter loss rates (5.3 +/- 2.3% per year in AD v 0.9 +/- 0.9% per year in controls) were faster in the left hemisphere (p < 0.029) than the right. Transient barriers to disease progression appeared at limbic/frontal boundaries. This degenerative sequence, observed in vivo as it developed, provides the first quantitative, dynamic visualization of cortical atrophic rates in normal elderly populations and in those with dementia. PMID- 12574430 TI - Molecular evidence for the functional role of dopamine D3 receptor in the morphine-induced rewarding effect and hyperlocomotion. AB - The aim of the present study was to investigate the role of dopamine D(3) receptors in the rewarding effect and hyperlocomotion induced by a prototypical mu-opioid receptor agonist morphine using dopamine D(3) receptor knock-out mice. The mu-opioid receptor in the brain determined by the [tylosil-3,5-(3)H(N)]-[D Ala(2),N-MePhe(4),Gly-ol(5)]enkephalin binding assay was not significantly changed by a deletion of the dopamine D(3) receptor gene. Furthermore, we found that no significant differences in G-protein activation by morphine in the limbic forebrain and lower midbrain were noted between the two genotypes. These results suggest that the function of the mu-opioid receptor itself was not affected by a deletion of the dopamine D(3) receptor gene. To ascertain the morphine-induced rewarding effect in both genotypes, the conditioned place preference paradigm was performed. Deletion of the dopamine D(3) receptor gene resulted in a remarkable enhancement of the morphine-induced rewarding effect. Furthermore, knock-out mice with deletions of the dopamine D(3) receptor revealed a dramatic potentiation of morphine-induced hyperlocomotion. Under these conditions, a loss of the dopamine D(3) receptor gene had no effect on the basal levels of dopamine and the increased dopamine turnover by morphine in the limbic forebrain. These findings provide further evidence that dopamine D(3) receptor contributes to the postsynaptically negative modulation of the mesolimbic dopaminergic pathway that is associated with the rewarding effect and hyperlocomotion through the stimulation of mu-opioid receptors induced by morphine in the mouse. PMID- 12574431 TI - Selective impairment of hippocampal gamma oscillations in connexin-36 knock-out mouse in vivo. AB - The physiological roles of neuronal gap junctions in the intact brain are not known. The recent generation of the connexin-36 knock-out (Cx36 KO) mouse has offered a unique opportunity to examine this problem. Recent in vitro recordings in Cx36 KO mice suggested that Cx36 gap junction contributes to various oscillatory patterns in the theta (approximately 5-10 Hz) and gamma (approximately 30-80 Hz) frequency ranges and affects certain aspects of high frequency (>100 Hz) patterns. However, the relevance of these pharmacologically induced patterns to the intact brain is not known. We recorded field potentials and unit activity in the CA1 stratum pyramidale of the hippocampus in the behaving wild-type (WT) and Cx36 KO mice. Fast-field "ripple" oscillations (140 200 Hz) were present in both WT and KO mice and did not differ significantly in power, intraepisode frequency, or probability of occurrence. Thus, fast-field oscillations either may not require electrical synapses or may be mediated by a hitherto unknown class of gap junctions. Theta oscillations, recorded during either wheel running or rapid eye movement sleep, were not different either. However, the power in the gamma frequency band and the magnitude of theta-phase modulation of gamma power were significantly decreased in KO mice compared with WT controls during wheel running. This suggests that Cx36 interneuronal gap junctions selectively contribute to gamma oscillations. PMID- 12574432 TI - Corticotropin releasing hormone type 2 receptors in the dorsal raphe nucleus mediate the behavioral consequences of uncontrollable stress. AB - Uncontrollable shock produces a constellation of behavioral changes that are not observed after equivalent escapable shock. These include interference with escape and potentiation of fear conditioning. The activation of corticotropin-releasing hormone (CRH) receptors within the caudal dorsal raphe nucleus (DRN) during inescapable tailshock (IS) has been shown to be critical for the development of these behavioral changes. CRH binds to two receptor subtypes, both of which are found in the DRN. The present set of studies examined which CRH receptor subtype mediates the effects of IS. Intra-DRN administration of the CRH(2) receptor antagonist anti-sauvagine-30 before IS dose-dependently blocked IS-induced behavioral changes; the CRH(1) receptor antagonist 2-methyl-4-(N-propyl-N cycloproanemethylamino)-5-chloro-6-(2,4,6-trichloranilino)pyrimidine (NBI27914), administered in the same manner, did not. Moreover, the highly selective CRH(2) receptor agonist urocortin II (Ucn II) dose-dependently caused behavioral changes associated with IS in the absence of shock. Ucn II was effective at doses 100 fold lower than those previously required for CRH. The relationship between CRH(2) receptors and DRN 5-HT is discussed. PMID- 12574433 TI - Spinal glia and proinflammatory cytokines mediate mirror-image neuropathic pain in rats. AB - Mirror-image allodynia is a mysterious phenomenon that occurs in association with many clinical pain syndromes. Allodynia refers to pain in response to light touch/pressure stimuli, which normally are perceived as innocuous. Mirror-image allodynia arises from the healthy body region contralateral to the actual site of trauma/inflammation. Virtually nothing is known about the mechanisms underlying such pain. A recently developed animal model of inflammatory neuropathy reliably produces mirror-image allodynia, thus allowing this pain phenomenon to be analyzed. In this sciatic inflammatory neuropathy (SIN) model, decreased response threshold to tactile stimuli (mechanical allodynia) develops in rats after microinjection of immune activators around one healthy sciatic nerve at mid-thigh level. Low level immune activation produces unilateral allodynia ipsilateral to the site of sciatic inflammation; more intense immune activation produces bilateral (ipsilateral + mirror image) allodynia. The present studies demonstrate that both ipsilateral and mirror-image SIN-induced allodynias are (1) reversed by intrathecal (peri-spinal) delivery of fluorocitrate, a glial metabolic inhibitor; (2) prevented and reversed by intrathecal CNI-1493, an inhibitor of p38 mitogen activated kinases implicated in proinflammatory cytokine production and signaling; and (3) prevented or reversed by intrathecal proinflammatory cytokine antagonists specific for interleukin-1, tumor necrosis factor, or interleukin-6. Reversal of ipsilateral and mirror-image allodynias was rapid and complete even when SIN was maintained constantly for 2 weeks before proinflammatory cytokine antagonist administration. These results provide the first evidence that ipsilateral and mirror-image inflammatory neuropathy pain are created both acutely and chronically through glial and proinflammatory cytokine actions. PMID- 12574434 TI - Selective disruption of stimulus-reward learning in glutamate receptor gria1 knock-out mice. AB - Glutamatergic neurotransmission via AMPA receptors has been an important focus of studies investigating neuronal plasticity. AMPA receptor glutamate receptor 1 (GluR1) subunits play a critical role in long-term potentiation (LTP). Because LTP is thought to be the cellular substrate for learning, we investigated whether mice lacking the GluR1 subunit [gria1 knock-outs (KO)] were capable of learning a simple cue-reward association, and whether such cues were able to influence motivated behavior. Both gria1 KO and wild-type mice learned to associate a light/tone stimulus with food delivery, as evidenced by their approaching the reward after presentation of the cue. During subsequent testing phases, gria1 KO mice also displayed normal approach to the cue in the absence of the reward (Pavlovian approach) and normal enhanced responding for the reward during cue presentations (Pavlovian to instrumental transfer). However, the cue did not act as a reward for learning a new behavior in the KO mice (conditioned reinforcement). This pattern of behavior is similar to that seen with lesions of the basolateral nucleus of the amygdala (BLA), and correspondingly, gria1 KO mice displayed impaired acquisition of responding under a second-order schedule. Thus, mice lacking the GluR1 receptor displayed a specific deficit in conditioned reward, suggesting that GluR1-containing AMPA receptors are important in the synaptic plasticity in the BLA that underlies conditioned reinforcement. Immunostaining for GluR2/3 subunits revealed changes in GluR2/3 expression in the gria1 KOs in the BLA but not the central nucleus of the amygdala (CA), consistent with the behavioral correlates of BLA but not CA function. PMID- 12574435 TI - Seasonal plasticity of peripheral auditory frequency sensitivity. AB - Female midshipman fish (Porichthys notatus) use the auditory sense to detect and locate vocalizing males during the breeding season. Detection of conspecific vocal signals is essential to their reproductive success and can evoke strong phonotactic responses in gravid females but not in spent females that have released all of their eggs. Here, we test the hypothesis that seasonal variation in reproductive state affects the neurophysiological response properties of the peripheral auditory system in female midshipman fish. Iso-intensity responses of eighth nerve afferents from the sacculus, the main auditory end organ of the inner ear, to individual tones were measured for spike rate and vector strength (VS) of synchronization. Most auditory saccular units in reproductive, summer females showed robust temporal encoding up to 340 Hz, whereas nonreproductive winter females showed comparable encoding only up to 100 Hz. The dramatic upward shift in temporal encoding among summer fish was paralleled by increases in best frequency (BF), maximum evoked spike rate at BF, VS values at BF, and the percentage of units that showed significant VS to iso-intensity tones >140 Hz. Reproductive summer females were best suited to encode the higher harmonic components of male advertisement calls. This first demonstration of a natural cyclicity in peripheral auditory frequency sensitivity among vertebrates may represent, in this case, an adaptive plasticity of the female midshipman's auditory system to enhance the acquisition of auditory information needed for mate identification and localization during the breeding season. PMID- 12574436 TI - Adaptive plasticity in the auditory thalamus of juvenile barn owls. AB - Little is known about the capacity of the thalamus for experience-dependent plasticity. Here, we demonstrate adaptive changes in the tuning of auditory thalamic neurons to a major category of sound localization cue, interaural time differences (ITDs), in juvenile barn owls that experience chronic abnormal hearing. Abnormal hearing was caused by a passive acoustic filtering device implanted in one ear that altered the timing and level of sound differently at different frequencies. Experience with this device resulted in adaptive, frequency-dependent shifts in the tuning of thalamic neurons to ITD that mimicked the acoustic effects of the device. Abnormal hearing did not alter ITD tuning in the central nucleus of the inferior colliculus, the primary source of input to the auditory thalamus. Therefore, the thalamus is the earliest stage in the forebrain pathway in which this plasticity is expressed. A visual manipulation, chronic prismatic displacement of the visual field, which causes adaptive changes in ITD tuning at higher levels in the forebrain, had no effect on thalamic ITD tuning. The results demonstrate that, during the juvenile period, auditory experience shapes neuronal response properties in the thalamus in a frequency specific manner and suggest that this thalamic plasticity is driven by self organizational forces and not by visual instruction. PMID- 12574437 TI - Genetic vasopressin deficiency facilitates performance of a lateralized reaction time task: altered attention and motor processes. AB - Brattleboro rats are a variety of the outbred Long-Evans strain that possess a single nucleotide deletion in the second exon of the arginine vasopressin gene, resulting in the synthesis of an altered protein that does not enter the normal secretory pathway. Rats heterozygous (di/+) for the deletion have a partial vasopressin deficiency and exhibit a variety of behavioral and neurochemical alterations compared with normal wild-type Long-Evans rats, which provide evidence for a CNS function for vasopressin. Here, we examined the acquisition and performance of a test of visuospatial attention by di/+ Brattleboro rats and their wild-type Long-Evans control counterparts. Surprisingly, di/+ rats exhibited superior performance of the task compared with wild-type controls; performance differences included greater accuracy of detection of visual target stimuli, faster overall reaction times, and fewer trial omissions. Di/+ rats also exhibited more approaches to the reinforcer receptacle at nonreinforcement times. These results indicate that alterations of vasopressin signaling result in a clear cognitive phenotype, including faster motor initiations and superior choice accuracy in a test of visual attention. PMID- 12574439 TI - Activity of different classes of neurons of the motor cortex during locomotion. AB - This study examines the activity of different classes of neurons of the motor cortex in the rabbit during two locomotion tasks: a simple (on a flat surface) and a complex (overstepping a series of barriers) locomotion. Four classes of efferent neurons were studied: corticocortical (CC) neurons with ipsilateral projection (CCIs), those with contralateral projection (CCCs), descending corticofugal neurons of layer V (CF5s), and those of layer VI (CF6s). In addition, one class of inhibitory interneurons (SINs) was investigated. CF5 neurons and SINs were the only groups that were strongly active during locomotion. In most of these neurons a clear-cut modulation of discharge in the locomotion rhythm was observed. During simple locomotion, CF5s and SINs were preferentially active in opposite phases of the step cycle, suggesting that SINs contribute to formation of the step-related pattern of CF5s. Transition from simple to complex locomotion was associated with changes of the discharge pattern of the majority of CF5 neurons and SINs. In contrast to CF5 neurons, other classes of efferent neurons (CCI, CCC, CF6) were much less active during both simple and complex locomotion. That suggests that CC interactions, both within a hemisphere (mediated by CCIs) and between hemispheres (mediated by CCCs), as well as corticothalamic interactions via CF6 neurons are not essential for motor coordination during either simple or complex locomotion tasks. PMID- 12574438 TI - Respiratory and telencephalic modulation of vocal motor neurons in the zebra finch. AB - Birdsong, like speech, involves coordinated vocal and respiratory activity achieved under telencephalic control. The avian vocal organ, or syrinx, is innervated by motor neurons (MNs) in the tracheosyringeal part of the hypoglossal nucleus (XIIts) that receive their synaptic input from medullary respiratory areas and telencephalic song control areas. Despite the importance of XIIts MNs to learned vocalizations, little is known about their intrinsic electrical properties or their synaptic inputs. Therefore, we made in vitro and in vivo intracellular recordings from XIIts MNs in adult male zebra finches to characterize their intrinsic properties and their synaptic modulation by respiratory and telencephalic areas. In vitro, electrical stimulation of ipsilateral or contralateral medullary respiratory areas (RAm) routinely evoked glycine receptor-mediated inhibition in XIIts. With inhibition blocked, similar stimulation evoked excitatory synaptic responses capable of driving sustained MN firing that was mediated partly by NMDA receptors. These inhibitory and excitatory inputs likely arise from RAm neurons, because chemical or electrical stimulation of RAm evoked similar responses in XIIts. In vivo, XIIts MNs displayed rhythmical, expiratory-related activity. EPSPs were pronounced at expiratory onset, but IPSPs were not apparent during inspiration, although XIIts MN firing was suppressed. However, hyperpolarizations as well as excitation were evoked by playback of the bird's own song, a stimulus that potently excites the telencephalic song nucleus that innervates XIIts. These findings illuminate functional properties of the songbird's brainstem circuitry and its specific activation by telencephalic inputs, which could coordinate vocal and respiratory activity during singing. PMID- 12574441 TI - The promise of translational physiology. PMID- 12574442 TI - Progressive adaptation of the soleus H-reflex with daily training at walking backward. AB - When untrained subjects walk backward on a treadmill the amplitude of the soleus H-reflex in midswing is equal to or exceeds the value in stance. This is a surprising result because during the swing phase of backward walking the soleus is inactive and its antagonist, the tibialis anterior, is active. We suggested that the high amplitude of the soleus H-reflex in late swing reflects task uncertainties, such as estimating the moment of foot contact with the ground and losing balance. In support of this idea we show that when untrained subjects held on to handrails the unexpected high-amplitude H-reflex during midswing was no longer present. We therefore asked whether daily training at this task without grasping the handrails would adaptively modify the H-reflex modulation pattern. In this event, within 10 days of training for 15 min daily, the anticipatory reflex activity at the beginning of training was gradually abated as the subjects reported gaining confidence at the task. However, when adapted subjects were made to walk backward with their eyes shut, the anticipatory reflex activity in midswing returned immediately. The reflex changes as a result of training were not due to changes in the motor activity or kinematics; they are likely part of the motor program controlling backward walking. This adaptive phenomenon may prove to be a useful model for studying the neural mechanisms of motor learning and adaptive plasticity in humans and may be relevant to rehabilitation programs for neurological patients. PMID- 12574443 TI - Membrane properties related to the firing behavior of zebrafish motoneurons. AB - The physiological and pharmacological properties of the motoneuron membrane and action potential were investigated in larval zebrafish using whole cell patch current-clamp recording techniques. Action potentials were eliminated in tetrodotoxin, repolarized by tetraethylammonium (TEA) and 3,4-diaminopyridine (3,4-AP)-sensitive potassium conductances, and had a cobalt-sensitive, high threshold calcium component. Depolarizing current injection evoked a brief (approximately 10-30 ms) burst of action potentials that was terminated by strong, outwardly rectifying voltage-activated potassium and calcium-dependent conductances. In the presence of intracellular cesium ions, a prolonged plateau potential often followed brief depolarizations. During larval development (hatching to free-swimming), the resting membrane conductance increased in a population of motoneurons, which tended to reduce the apparent outward rectification of the membrane. The conductances contributing to action potential burst termination are hypothesized to play a role in patterning the synaptically driven motoneuron output in these rapidly swimming fish. PMID- 12574444 TI - Importance of cutaneous feedback in maintaining a secure grip during manipulation of hand-held objects. AB - Previous research has shown that grip and load forces are modulated simultaneously during manipulation of a hand-held object. This close temporal coupling suggested that both forces are controlled by an internal model within the CNS that predicts the changes in tangential force on the fingers. The objective of the present study was to examine how the internal model would compensate for the loss of cutaneous sensation through local anesthesia of the index and thumb. Ten healthy adult subjects (5 men and 5 women aged 20-57 yr) were asked to grasp, lift, and hold stationary, a 250 g object for 20 s. Next, the subjects were asked to perform vertical oscillatory movements over a distance of 20 cm at a rate of 1.0 Hz for 30 s. Eleven trials were performed with intact sensation, and 11 trials after a local ring-block anesthesia of the index and thumb with bupivacain (5 mg/ml). During static holding, loss of cutaneous sensation produced a significant increase in the safety margin. However, the grip force declined significantly over the 20-s static hold period. During oscillatory arm movements, grip and load forces were continuously modulated together in a predictive manner as suggested by Flanagan and Wing. Again, the grip force declined over the 30-s movement, and 7/10 subjects dropped the object at least once. With intact sensation, the object was never dropped; but with the fingers anesthetized, it was dropped on 36% of the trials, and a significant slip occurred on a further 12%. The mean correlation between the grip and load forces for all subjects deteriorated from 0.71 with intact sensation to 0.48 after digital anesthesia. However, a cross-correlation calculated between the grip and load forces indicated that the phase lag was approximately zero both with and without digital anesthesia. Taken together, the data from the present study suggest that cutaneous afferents are required for setting and maintaining the background level of the grip force in addition to their phasic slip-detection function and their role in adapting the grip force/load force ratio to the friction on initial contact with an object. Finally, at a more theoretical level, they correct and maintain an internal model of the physical properties of hand held objects. PMID- 12574445 TI - The effects of digital anesthesia on force control using a precision grip. AB - A total of 20 right-handed subjects were asked to perform a grasp-lift-and-hold task using a precision grip. The grasped object was a one-degree-of-freedom manipuladum consisting of a vertically mounted linear motor capable of generating resistive forces to simulate a range of object weights. In the initial study, seven subjects (6 women, 1 man; ages 24-56 yr) were first asked to lift and hold the object stationary for 4 s. The object presented a metal tab with two different surface textures and offered one of four resistive forces (0.5, 1.0, 1.5, and 2.0 N). The lifts were performed both with and without visual feedback. Next, the subjects were asked to perform the same grasping sequence again after ring block anesthesia of the thumb and index finger with mepivacaine. The objective was to determine the degree to which an internal model obtained through prior familiarity might compensate for the loss of cutaneous sensation. In agreement with previous studies, it was found that all subjects applied significantly greater grip force after digital anesthesia, and the coordination between grip and load forces was disrupted. It appears from these data, that the internal model alone is insufficient to completely compensate for the loss of cutaneous sensation. Moreover, the results suggest that the internal model must have either continuous tonic excitation from cutaneous receptors or at least frequent intermittent reiteration to function optimally. A subsequent study performed with 10 additional subjects (9 women, 1 man; ages 24-49 yr) indicated that with unimpaired cutaneous feedback, the grasping and lifting forces were applied together with negligible forces and torques in other directions. In contrast, after digital anesthesia, significant additional linear and torsional forces appeared, particularly in the horizontal and frontal planes. These torques were thought to arise partially from the application of excessive grip force and partially from a misalignment of the two grasping fingers. These torques were further increased by an imbalance in the pressure exerted by the two opposing fingers. Vision of the grasping hand did not significantly correct the finger misalignment after digital anesthesia. Taken together, these results suggest that mechanoreceptors in the fingertips signal the source and direction of pressure applied to the skin. The nervous system uses this information to adjust the fingers and direct the pinch forces optimally for grasping and object manipulation. PMID- 12574446 TI - Protein phosphatases mediate depotentiation induced by high-intensity theta-burst stimulation. AB - We have previously reported that varying stimulus intensity produces qualitatively different types of synaptic plasticity in area CA1 of hippocampal slices: brief low-intensity (LI) theta-burst (TB) stimuli induce long-term potentiation (LTP), but if the stimulus intensity is increased (to mimic conditions that may exist during seizures), LTP is not induced; instead, high intensity (HI) TB stimuli erase previously induced LTP ("TB depotentiation"). We now have explored the mechanisms underlying TB depotentiation using extracellular field recordings with pharmacological manipulations. We found that TB depotentiation was blocked by okadaic acid and calyculin A (inhibitors of serine/threonine protein phosphatases PP1 and PP2A), FK506 (a specific blocker of calcineurin, a Ca(2+)/calmodulin (CaM) protein phosphatase), and 8-Br-cAMP (an activator of protein kinase A) with 3-isobutyl-1-methylxanthine (IBMX, a phosphodiesterase inhibitor). These results suggest that protein phosphatase pathways are involved in the TB depotentiation similar to other type of down regulating synaptic plasticity such as low-frequency stimulation (LFS)-induced long-term depression (LTD) and depotentiation in the rat hippocampus. However, TB depotentiation and LFS depotentiation could have differential functional significance. PMID- 12574447 TI - Glycine tranporter-1 blockade potentiates NMDA-mediated responses in rat prefrontal cortical neurons in vitro and in vivo. AB - The N-methyl-D-aspartate (NMDA) receptor (NMDA-R) has pivotal roles in neural development, learning, memory, and synaptic plasticity. Functional impairment of NMDA-R has been implicated in schizophrenia. NMDA-R activation requires glycine to act on the glycine-B (GlyB) site of the NMDA-R as an obligatory co-agonist with glutamate. Extracellular glycine near NMDA-R is regulated effectively by a glial glycine transporter (GlyT1). Using whole-cell voltage-clamp recordings in prefrontal cortex (PFC) slices, we have shown that exogenous GlyB site agonists glycine and D-serine, or a specific GlyT1 inhibitor N[3-(4'-fluorophenyl)-3-(4' phenylphenoxy)propyl]sarcosine (NFPS) in the presence of exogenous glycine (10 microM), potentiated synaptically evoked NMDA excitatory postsynaptic currents (EPSCs) in vitro. Furthermore, in urethan-anesthetized rats, microiontophoretic NMDA pulses excite single PFC neurons. When these responses were blocked by approximately 50% to approximately 90% on continuous iontophoretic application of the GlyB site, antagonist (+)HA-966, intravenous NFPS (5 mg/kg), or a GlyB site agonist D-serine (50 mg/kg iv) reversed this (+)HA-966 block. NFPS may elevate endogenous glycine levels sufficiently to displace (+)HA-966 from the GlyB sites of the NMDA-R, thus enabling reactivation of the NMDA-Rs by iontophoretic NMDA applications. D-Serine (50-100 mg/kg iv) or NFPS (1-2 mg/kg iv) alone also augmented NMDA-evoked excitatory responses. These data suggest that direct GlyB site stimulation by D-serine, or blockade of GLYT1 to elevate endogenous glycine to act on unsaturated GlyB sites on NMDA-Rs, potentiated NMDA-R-mediated firing responses in rat PFC. Hence, blockade of GlyT1 to elevate glycine near the NMDA-R may activate hypofunctional NMDA-R, which has been implicated to play a critical role in the pathophysiology of schizophrenia. PMID- 12574448 TI - Efficacy and short-term plasticity at GABAergic synapses between Purkinje and cerebellar nuclei neurons. AB - Although the entire output of the cerebellar cortex is conveyed to the deep cerebellar nuclei neurons (DCNs) via the GABAergic synapses established by Purkinje cells (PCs), very little is known about the strength and dynamic properties of PC-DCN connections. Here we show that activation of PC-DCN unitary connections induced large conductance changes (11.7 nS) in DCNs recorded in whole cell patch configuration in acute slices, suggesting that activity of single PCs might significantly affect the output of its target neurons. Based on the large unitary quantal content (18) inferred from calculations of PC-DCN quantal size (0.65 nS) and the near absence of failures in synaptic transmission during control conditions, we conclude that PC-DCN connections are highly multi-sited. The analysis of dynamic properties of PC-DCN synapses demonstrated remarkable paired pulse depression (PPD), maximal at short intervals (paired pulse ratio of 0.15 at 7-ms interval). We provide evidence that PPD is presynaptic in origin and release-independent. In addition, multiple pulse stimulation revealed that PC-DCN synapses exhibited larger sensitivity to dynamic than to steady signals. We postulate that the, otherwise paradoxical, combination of marked short-term depression with strong multi-sited connections is optimal to transfer dynamic information at unitary level by performing spatial average of release probability across the numerous release sites. This feature could enable these synapses to encode presynaptic time-varying signals of single PCs as moment-to-moment changes in synaptic strength, a capacity well suited to the postulated role of cerebellum in control of temporal aspects of motor or cognitive behaviors. PMID- 12574449 TI - Differential sensitization of amygdala neurons to afferent inputs in a model of arthritic pain. AB - Pain is associated with negative affect such as anxiety and depression. The amygdala plays a key role in emotionality and has been shown to undergo neuroplastic changes in models of affective disorders. Many neurons in the central nucleus of the amygdala (CeA) are driven by nociceptive inputs, but the role of the amygdala in persistent pain states is not known. This study is the first to address nociceptive processing by CeA neurons in a model of prolonged pain. Extracellular single-unit recordings were made from 41 CeA neurons in anesthetized rats. Each neuron's responses to brief mechanical stimulation of joints, muscles, and skin and to cutaneous thermal stimuli were recorded. Background activity, receptive field size, and threshold were mapped, and stimulus-response functions were constructed. These parameters were measured repeatedly before and after induction of arthritis in one knee by intraarticular injections of kaolin and carrageenan. Multireceptive (MR) amygdala neurons (n = 20) with excitatory input from the knee joint responded more strongly to noxious than to innocuous mechanical stimuli of deep tissue (n = 20) and skin (n = 11). After induction of arthritis, 18 of 20 MR neurons developed enhanced responses to mechanical stimuli and expansion of receptive field size. These changes occurred with a biphasic time course (early peak: 1-1.5 h; persistent plateau phase: after 3-4 h). Responses to thermal stimuli did not change (7 of 7 neurons), but background activity (16 of 18 neurons) and electrically evoked orthodromic activity (11 of 12 neurons) increased in the arthritic state. Nociceptive specific (NS) neurons (n = 13) showed no changes of their responses to mechanical, thermal, and electrical stimulation after induction of arthritis. A third group of neurons did not respond to somesthetic stimuli under control conditions (noSOM neurons; n = 8) but developed prolonged responses to mechanical, but not thermal, stimuli in arthritis (5 of 8 neurons). These data suggest that prolonged pain is accompanied by enhanced responsiveness of a subset of CeA neurons. Their sensitization to mechanical, but not thermal, stimuli argues against a nonspecific state of hyperexcitability. MR neurons could serve to integrate and evaluate information in the context of prolonged pain. Recruitment of noSOM neurons increases the gain of amygdala processing. NS neurons preserve the distinction between nociceptive and nonnociceptive inputs. PMID- 12574450 TI - Substance P depresses excitatory synaptic transmission in the nucleus accumbens through dopaminergic and purinergic mechanisms. AB - Substance P (SP) is an undecapeptide that is co-localized with conventional transmitters in the nucleus accumbens (NAc). Its neurochemical and behavioral effects resemble those of cocaine and amphetamine. How SP accomplishes these effects is not known, partly because its cellular and synaptic effects are not well characterized. Using whole cell and nystatin-perforated patch recording in rat forebrain slices, we show here that SP, an excitatory neuropeptide, depresses evoked excitatory postsynaptic currents (EPSCs) and potentials (EPSPs) in NAc through intermediate neuromodulators. SP caused a partially reversible, dose dependent decrease in evoked EPSCs. This effect was mimicked by a neurokinin-1 (NK1) receptor-selective agonist, [Sar(9), Met (O(2))(11)]-SP and blocked by a NK1 receptor-selective antagonist, L 732 138. Both the SP- and [Sar(9), Met (O(2))(11)]-SP-induced synaptic depressions were accompanied by increases in paired pulse ratio (PPR), effects that were also blocked by L 732 138. In contrast to its effect on PPR, SP did not produce significant changes in the holding current, input resistance, EPSC decay rate (tau), and steady-state I-V curves of the recorded cells. The SP-induced synaptic depressions were prevented by dopamine receptor blockade using SCH23390 and haloperidol, but not by sulpiride. In addition, the SP-induced synaptic depression was blocked by an adenosine A1 receptor blocker 8-cyclopentyltheophylline (8-CPT) but not the N methyl-D-aspartate (NMDA) receptor antagonist D-APV. These data show that SP, by activating presynaptic NK1 receptors, depresses excitatory synaptic transmission indirectly by enhancing extracellular dopamine and adenosine levels. Since the cellular and synaptic effects of SP resemble those of cocaine and amphetamine, it may serve as an endogenous psychogenic peptide. PMID- 12574451 TI - Summation of forces from multiple motor units in the cat soleus muscle. AB - Nearly all muscle models and most motor control concepts assume that forces from individual muscle fibers and motor units sum in an additive manner once effects of in-series tendon compliance are taken into account. Due to the numerous mechanical linkages between individual fibers, though, it is unclear whether this assumption is warranted. This work examined motor unit force summation over a wide range of muscle forces in the cat soleus. Nonadditive summation implies a nonlinear summation of motor unit forces. Summation nonlinearities were quantified during interactions of 10 individual motor units and 4 motor unit bundles containing approximately 10 units each. These protocols allowed motor unit force summation to be examined from approximately 0 to 25% of tetanic muscle force. Nonlinear summation was assessed by comparing the actual forces to the algebraic sum of individual units and bundles stimulated in isolation. Superadditive summation meant that the actual force exceeded the algebraic sum, whereas subadditive summation meant that the actual force was smaller than the algebraic sum. Experiments tested the hypothesis that superadditive summation occurs at low force levels when few motor units are recruited, whereas subadditive summation prevails above 10% of tetanic force. Results were consistent with this hypothesis. As in previous studies, nonlinear summation in the soleus was modest, but a clear transition from predominately superadditive to predominantly subadditive summation occurred in the range of 6-8% of tetanic force. The largest nonlinearities were transient and appeared at the onset of recruitment and derecruitment of groups of motor units. The results are discussed in terms of the mechanical properties of the connective tissue forming the tendon and linking muscle fibers. PMID- 12574452 TI - Quantification of cardiac sac network effects on a movement-related parameter of pyloric network output in the lobster. AB - Cardiac sac network activity (cycle period tens of seconds to minutes) has long been known to alter pyloric network activity (cycle period approximately 1 s), but these effects have not been quantified. Some pyloric muscles extract cardiac sac timed variations in pyloric motor neuron firing, and consequently produce cardiac sac timed movements even though no cardiac sac neurons innervate them. Determining pyloric behavior therefore requires detailed description of cardiac sac effects on pyloric neural output. Pyloric muscle activity correlates well with motor neuron overall spike frequency (OSF, number of spikes per burst divided by cycle period). We therefore quantified the effects of cardiac sac activity on the OSF of all pyloric neurons in the lobster, Panulirus interruptus. The ventricular dilator (VD) neuron had a biphasic response, with its OSF first increasing and then decreasing during cardiac sac bursts. Lateral pyloric (LP) neuron OSF decreased during cardiac sac activity. The pyloric (PY) neurons had two responses, with OSF either decreasing or increasing just after the beginning of cardiac sac activity. The pyloric dilator (PD) neurons had a triphasic response, with OSF increasing slightly at the beginning of cardiac sac activity, decreasing during the cardiac sac burst, and strongly increasing after cardiac sac activity ended. The inferior cardiac (IC) neuron had a biphasic response, with OSF decreasing at the beginning of cardiac sac activity and strongly increasing when cardiac sac activity ceased. These data provide the quantitative description of cardiac sac effects on pyloric activity necessary to predict pyloric movement from pyloric neural output. PMID- 12574453 TI - Retinal bipolar neurons express the cyclic nucleotide-gated channel of cone photoreceptors. AB - Cyclic nucleotide-gated (CNG) channels link intracellular cyclic nucleotides to changes in membrane ionic conductance in a variety of physiological contexts. In the retina, in addition to their central role in phototransduction, CNG channels may be involved in nitric oxide signaling in bipolar neurons or in the hyperpolarizing synaptic response to glutamate in ON-type (depolarizing) bipolar cells. Despite their potential physiological significance, however, expression of CNG channels has not yet been demonstrated in bipolar cells. To identify CNG channel subtypes in retinal bipolar neurons, we used single-cell molecular biological techniques in morphologically distinctive ON bipolar cells from goldfish retina. Both single-cell in situ hybridization and single-cell RT-PCR demonstrated in ON bipolar cells the presence of mRNA for the CNG channel subtype that is also found in cone photoreceptors. Other bipolar cells, which likely represent OFF cells, did not express the cone CNG channel. Thus the CNG channel of cone photoreceptors is expressed in ON bipolar cells, where it may be involved in physiological responses to nitric oxide, or in the sign-inverting glutamatergic synapse that gives rise to the ON visual pathway. PMID- 12574454 TI - G protein signaling in a neuronal network is necessary for rhythmic motor pattern production. AB - G protein-coupled receptors are widely recognized as playing important roles in mediating the actions of extrinsic neuromodulatory inputs to motor networks. However, the potential for their direct involvement in rhythmic motor pattern generation has received considerably less attention. Results from this study indicate that G protein signaling appears to be integral to the operation of the central pattern generator (CPG) underlying the escape swim of the mollusk Tritonia diomedea. Blocking G protein signaling in a single CPG neuron, cerebral neuron C2, with intracellular iontophoresis of the guanine nucleotide analogue guanosine 5'-O-(2-thiodiphosphate) (GDP-beta-S), prevented the production of the swim motor program. Moreover, tonic activation of G protein signaling in this neuron by iontophoresis of the GTP analogues guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S) and 5'-guanylyl-imidodiphosphate also inhibited motor pattern production. The possible sites of action of these guanine nucleotide analogues were examined to assess potential mechanisms by which they interfered with motor pattern production. Intracellular iontophoresis of GDP-beta-S into C2 did not affect C2 basal synaptic strength. However, it did reduce heterosynaptic facilitation of C2 synapses caused by the dorsal swim interneurons (DSIs), a set of serotonergic swim CPG neurons. In contrast, GTP-gamma-S directly enhanced C2 synaptic strength onto DFN, mimicking the neuromodulatory effect of the DSIs. GDP beta-S, but not the GTP analogues, decreased C2 excitability, whereas both GTP analogues, but not GDP-beta-S, blocked the ability of DSI stimulation to increase C2 excitability. The decrease in C2 excitability caused by GDP-beta-S is not likely to be responsible for the inhibition of the swim motor pattern because decreasing C2 firing rate, by injecting hyperpolarizing current, did not prevent the production of the rhythmic motor pattern. Taken together, these data suggest that G protein signaling is a necessary and integral component of the escape swim CPG in Tritonia and that G protein signaling mediates DSI heterosynaptic facilitation of C2 but may not mediate the DSI-evoked enhancement of C2 excitability. PMID- 12574455 TI - Neural pathways between sacrocaudal afferents and lumbar pattern generators in neonatal rats. AB - Projections of sacrocaudal afferents (SCA) onto lumbar pattern generators were studied in isolated spinal cords of neonatal rats. A locomotor-like pattern could be produced by SCA stimulation in the majority of the preparations. The SCA induced lumbar rhythm was abolished after blocking synaptic transmission in the sacrococcygeal (SC) cord by bathing its segments in a low-calcium, high-magnesium artificial cerebrospinal fluid and restored when the synaptic block was alleviated by local application of calcium onto specific SC segments prior to SCA stimulation. Thus the SCA evoked lumbar rhythm involves synaptic activation of relay neurons in the SC cord. Functional activation of these relays depends on non-N-methyl-D-aspartate (NMDA) receptors because the lumbar rhythm was abolished when the non-NMDA receptor antagonist CNQX was added to the SC cord. By contrast, pharmacological block of the rhythmicity in the SC cord by specific antagonists of NMDA receptors and alpha1 and alpha2 adrenoceptors did not impair the SCA induced lumbar rhythm. Midsagittal splitting experiments of parts of the SC and lumbar cord revealed that crossed and uncrossed ascending/propriospinal pathways are coactivated by SCA stimulation. We suggest that these pathways ascend onto the thoracolumbar cord through the lateral, ventrolateral, and ventral funiculi, because a complete block of the lumbar rhythm could only be obtained with a bilateral interruption of all of these funiculi. The relevance of our findings to the neural control of the rhythmogenic networks in the spinal cord is discussed. PMID- 12574456 TI - Encoding of compressive stress during indentation by group III and IV muscle mechano-nociceptors in rat gracilis muscle. AB - The mechanical state encoded by group III and IV muscle afferents, putative mechano-nociceptors, during indentation was examined using an isolated muscle nerve preparation in a rat model. Gracilis muscle and its intact innervation were surgically removed from the medial thigh of the rat hindlimb and placed in a dish containing rodent synthetic interstitial fluid. The tendons of the muscle were coupled to an apparatus that could stretch and apply compression to the muscle. Using a standard teased-nerve preparation, the neural responses of single mechanically sensitive group III or IV afferents were identified. Afferents were classified as mechano-nociceptors on the basis of their graded response to noxious levels of compressive stress (or strain) as well as, in some cases, their polymodal response to noxious thermal stimuli. Mechano-nociceptors (n = 13) were stimulated using controlled compressive stress (10-30 kPa) or strain (40-80%) while simultaneously measuring displacement and force by compressing the muscle between a flat cylinder and a hard platform. Linear regression was used to evaluate the relationships between neural response and mechanical stress, force, strain, and displacement. The mean neural response (threshold: 1.1 +/- 0.4 kPa; sensitivity: 0.5 +/- 0.1 Hz/kPa; means +/- SE) was significantly and substantially more highly correlated with compressive stress than force, strain, or displacement. The data from this study support the hypothesis that muscle nociceptors stimulated by indentation encode compressive stress rather than force, strain, or displacement. PMID- 12574457 TI - Increased electrotonic coupling in spinal motoneurons after transient botulinum neurotoxin paralysis in the neonatal rat. AB - The effect of early postnatal blockade of neuromuscular transmission using botulinum neurotoxin (BoNT) type A on motoneuron gap junctional coupling was studied by means of intracellular recordings and biocytin labeling using the in vitro hemisected spinal cord preparation of neonatal rats. The somata of tibialis anterior (TA) motoneurons were retrogradely labeled at birth (P0) by intramuscular injection of fluorescent tracers. Two days later, BoNT was injected unilaterally into the TA muscle. The toxin blocked neuromuscular transmission for the period studied (P4-P7) as shown by tension recordings of the TA muscle. Retrograde horseradish peroxidase tracing in animals reared to adulthood demonstrated no significant cell death or changes in the soma size of BoNT treated TA motoneurons. Intracellular recordings were carried out in prelabeled control and BoNT-treated TA motoneurons from P4 to P7. Graded stimulation of the ventral root at subthreshold intensities elicited short-latency depolarizing (SLD) potentials that consisted of several discrete components reflecting electrotonic coupling between two or more motoneurons. BoNT treatment produced a significant increase (67%) in the maximum amplitude of the SLD and in the number of SLD components as compared with control (3.1 +/- 1.7 vs. 1.4 +/- 0.7; means +/ SD). The morphological correlates of electrotonic coupling were investigated at the light microscope level by studying the transfer of biocytin to other motoneurons and the putative sites of gap junctional interaction. The dye-coupled neurons clustered around the injected cell with close somato-somatic, dendro somatic and -dendritic appositions that might represent the sites of electrotonic coupling. The size of the motoneuron cluster was, on average, 2.2 times larger after BoNT treatment. Our findings demonstrate that a short-lasting functional disconnection of motoneurons from their target muscle delays motoneuron maturation by halting the elimination of gap junctional coupling that normally occurs during early postnatal development. PMID- 12574458 TI - Ethanol dual modulatory actions on spontaneous postsynaptic currents in spinal motoneurons. AB - Recently we have shown that acute ethanol (EtOH) exposure suppresses dorsal root evoked synaptic potentials in spinal motoneurons. To examine the synaptic mechanisms underlying the reduced excitatory activity, EtOH actions on properties of action potential-independent miniature excitatory and inhibitory postsynaptic currents (mEPSCs and mIPSCs) were studied in spinal motoneurons of newborn rats. Properties of mEPSCs generated by activation of N-methyl-D-aspartate receptors (NMDARs) and non-NMDA receptors and of mIPSCs mediated by glycine and gamma aminobutyric acid-A receptors (GlyR and GABA(A)R) were examined during acute exposure to 70 and 200 mM EtOH. In the presence of 70 mM EtOH, the frequency of NMDAR- and non-NMDAR-mediated mEPSCs decreased to 53 +/- 5 and 45 +/- 7% (means +/- SE) of control values, respectively. In contrast, the frequency of GlyR- and GABA(A)R-mediated mIPSCs increased to 138 +/- 15 and 167 +/- 23% of control, respectively. Based on the quantal theory of transmitter release, changes in the frequency of miniature currents are correlated with changes in transmitter release, suggesting that EtOH decreased presynaptic glutamate release and increased the release of both glycine and GABA. EtOH did not change the amplitude or rise and decay times of either mEPSCs or mIPSCs, indicating that the presynaptic changes were not associated with changes in the properties of postsynaptic receptors/channels. Acute exposure to 200 mM EtOH increased mIPSC frequency two- to threefold, significantly higher than the increase induced by 70 mM EtOH. However, the decrease in mEPSC frequency was similar to that observed in 70 mM EtOH. Those findings implied that the regulatory effect of EtOH on glycine and GABA release was dose-dependent. Exposure to the higher EtOH concentration had opposite actions on mEPSC and mIPSC amplitudes: it attenuated the amplitude of NMDAR- and non-NMDAR-mediated mEPSCs to ~80% of control and increased GlyR- and GABA(A)R-mediated mIPSC amplitude by ~20%. EtOH-induced changes in the amplitude of postsynaptic currents were not associated with changes in their basic kinetic properties. Our data suggested that in spinal networks of newborn rats, EtOH was more effective in modulating the release of excitatory and inhibitory neurotransmitters than changing the properties of their receptors/channels. PMID- 12574459 TI - Effects of echo intensity on Doppler-shift compensation behavior in horseshoe bats. AB - Echolocating horseshoe bats respond to flight-speed induced shifts in echo frequency by adjusting the frequency of subsequent calls. Under natural conditions, Doppler effects may force the frequency of a returning echo several kilohertz above the original emission frequency. By lowering subsequent call frequencies, the bat can return echo frequencies to within a narrow spectral bandwidth to which its highly specialized auditory system is most sensitive. While Doppler-shift compensation (DSC) behavior specifically refers to frequency compensation, other parameters of the returning echo, such as delay, duration, and interaural time and intensity differences have been shown to influence DSC performance. Understanding the nature of these influences has already led to a better appreciation of the neurophysiology of DSC. Here we provide a quantitative analysis of the effects of a prominent feature of the returning echo, its intensity, on DSC performance in horseshoe bats. Although DSC performance generally tolerates echo attenuation up to approximately 40 dB relative to the outgoing emission intensity, a systematic decline in DSC performance can be observed over this range. Generally, the effects of echo attenuation are characterized by a reduction in 1) the overall amount of compensation relative to the size of the shift in echo frequency and 2) the rate at which the bat responds to perceived echo shifts. These effects appear to be the consequence of a systematic shift in the range of echo frequencies capable of inducing DSC behavior. In particular, the reference frequency (the minimum shift in echo frequency that will elicit DSC behavior) appears to be highly sensitive to echo intensity. Every 10-dB reduction in echo intensity shifts the reference upward nearly 250 Hz. Our results indicate that, even at the highest intensity levels, relatively minor changes in echo intensity critically influence frequency compensation during normal DSC. We conclude with a discussion of how these results might impact echolocation behavior of horseshoe bats under natural and experimental conditions. PMID- 12574460 TI - Excitatory amino acid receptors of the electrosensory system: the NR1/NR2B N methyl-D-aspartate receptor. AB - The amino acid sequence of the N-methyl-D-aspartate (NMDA) receptor subunit NR2B from the brown ghost knife fish Apteronotus leptorhynchus has been determined and compared with the sequence of the murine NR2B. This comparison revealed high levels of sequence conservation throughout the ligand binding and membrane spanning segments. The functional properties of the NR1 and NR2B receptor complex were examined by coexpression in HEK cells. The recombinant AptNR1/NR2B receptors produced robust currents after stimulation with glutamate or NMDA in the presence of glycine. Measurements of the concentration dependencies for these agonists indicated that the agonist binding sites on the apteronotid receptor are highly conserved, with nearly identical agonist affinities to those of the murine NR1/NR2B receptor. The kinetic responses of the fish receptor were also highly conserved, with deactivation rates for the AptNR2B receptor matching those of the murine NR2B containing receptor. Evidently, most of the unique functional properties that reside in the NR2B receptor subunit have been well conserved in teleost NMDA receptors. On the other hand, the apteronitid receptor displayed a lowered sensitivity to voltage-dependent Mg(2+) block and a reduced affinity for the NR2B-specific noncompetitive antagonist ifenprodil. We conclude that the functional properties that result from the incorporation of the NR2B receptor in the NMDA receptor complex have been maintained since the evolutionary divergence of teleost and mammalian organisms. PMID- 12574461 TI - Fractal properties of sympathetic nerve discharge. AB - Fano factor analysis and dispersional analysis were used to characterize time series of single and multifiber spikes recorded from the preganglionic cervical sympathetic nerve and cardiac-related slow-wave activity of the whole postganglionic sympathetic vertebral nerve (VN) in anesthetized cats. Fluctuations in spike counts and interspike intervals for single preganglionic fibers proved to be fractal (i.e., time-scale invariant), as reflected by a power law relationship between indices of the variance of these properties and the window size used to make the measurements. Importantly, random shuffling of the data eliminated the power law relationships. Fluctuations in spike counts in preganglionic multifiber activity also were fractal, as were fluctuations in the height and of the area of cardiac-related slow waves recorded from the whole postganglionic VN. These fractal fluctuations were persistent (i.e., positively correlated), as reflected by a Hurst exponent significantly >0.5. Although fluctuations in the interval between cardiac-related VN slow waves were random, those in the interval between heart beats were fractal and persistent. These results demonstrate for the first time that apparently random fluctuations in sympathetic nerve discharge are, in fact, dictated by a complex deterministic process that imparts "long-term" memory to the system. Whether such time-scale invariant behavior plays a role in generating the fractal component of heart rate variability remains to be determined. PMID- 12574462 TI - Neocortical very fast oscillations (ripples, 80-200 Hz) during seizures: intracellular correlates. AB - Multi-site field potential and intracellular recordings from various neocortical areas were used to study very fast oscillations or ripples (80-200 Hz) during electrographic seizures in cats under ketamine-xylazine anesthesia. The animals displayed spontaneously occurring and electrically induced seizures comprising spike-wave complexes (2-3 Hz) and fast runs (10-20 Hz). Neocortical ripples had much higher amplitudes during seizures than during the slow oscillation preceding the onset of seizures. A series of experimental data from the present study supports the hypothesis that ripples are implicated in seizure initiation. Ripples were particularly strong at the onset of seizures and halothane, which antagonizes the occurrence of ripples, also blocked seizures. The firing of electrophysiologically defined cellular types was phase-locked with ripples in simultaneously recorded field potentials. This indicates that ripples during paroxysmal events are associated with a coordination of firing in a majority of neocortical neurons. This was confirmed with dual intracellular recordings. Based on the amplitude that neocortical ripples reach during paroxysmal events, we propose a mechanism by which neocortical ripples during normal network activity could actively participate in the initiation of seizures on reaching a certain threshold amplitude. This mechanism involves a vicious feedback loop in which very fast oscillations in field potentials are a reflection of synchronous action potentials, and in turn these oscillations help generate and synchronize action potentials in adjacent neurons through electrical interactions. PMID- 12574463 TI - Sympathetic modulation of acute cutaneous flare induced by intradermal injection of capsaicin in anesthetized rats. AB - Much of the acute cutaneous neurogenic inflammation after intradermal injection of capsaicin (CAP) in rats is mediated by dorsal root reflexes (DRRs), which cause the release of inflammatory agents from primary afferent terminals. Sympathetic efferents modulate neurogenic inflammation by interaction with primary afferent terminals. In this study, we examined if DRR-mediated flare after CAP injection is subject to sympathetic modulation. Changes in cutaneous blood flow on the plantar surface of the foot were measured using a laser Doppler flow meter. After CAP injection, cutaneous flare spread more than 20 mm away from the site of CAP injection. However, this CAP-induced flare was significantly reduced after surgical sympathectomy. Decentralization of postganglionic neurons did not affect the flare induced by CAP injection. If the foot of sympathectomized rats was pretreated with an alpha(1)-adrenoceptor agonist (phenylephrine) by intra-arterial injection, the spread of flare induced by CAP injection could be restored. However, if the spinal cord was pretreated with a GABA(A) receptor antagonist, bicuculline, to prevent DRRs, phenylephrine no longer restored the CAP-evoked flare. An alpha(2)-adrenoceptor agonist (UK14,304) did not affect the CAP-evoked flare in sympathectomized rats. In sympathetically intact rats, blockade of peripheral alpha(1)-adrenoceptors with terazosin profoundly reduced the flare induced by CAP injection, whereas blockade of peripheral alpha(2)-adrenoceptors by yohimbine did not obviously affect the flare. Therefore the pathogenesis of acute neurogenic inflammation in the intradermal CAP injection model depends in part on intact sympathetic efferents and alpha(1)-adrenoceptors. Peripheral alpha(1)-adrenoceptors thus modulate the ability of capsaicin sensitive afferents to evoke the release of inflammatory agents from primary afferents by DRRs. PMID- 12574464 TI - Differential processing of excitation by GABAergic gain modulation in canine caudal ventral respiratory group neurons. AB - The discharge frequency (F(n)) patterns of medullary respiratory premotor neurons are subject to potent tonic GABAergic gain modulation. Studies in other neuron types suggest that the synaptic input for tonic inhibition is located on the soma where it can affect total neuronal output. However, our preliminary data suggested that excitatory responses elicited by highly local application of glutamate receptor agonists are not gain modulated. In addition, modulation of the amplitude of spike afterhyperpolarizations can gain modulate neuronal output, and this mechanism is located near the spike initiation zone and/or soma. The purpose of this study was to determine if these two gain-modulating mechanisms have different functional locations on the somatodendritic membrane of bulbospinal inspiratory and expiratory neurons. Four-barrel micropipettes were used for extracellular single-neuron recording and pressure ejection of drugs in decerebrate, paralyzed, ventilated dogs. The net increases in F(n) due to repeated short-duration picoejections of the glutamate receptor agonist, alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), was quantified before and during locally induced antagonism of GABA(A) receptors by bicuculline or small-conductance, calcium-activated potassium channels by apamin. The AMPA induced net increases in F(n) were not significantly altered by BIC, although it produced large increases in the respiratory-related activity. However, the AMPA induced net responses were amplified in accordance with the gain increase of the respiratory-related activity by apamin. These findings suggest that GABAergic gain modulation may be functionally isolated from the soma/spike initiation zone, e.g., located on a dendritic shaft. This could allow other behavioral signals requiring strong neuronal activation (e.g., coughing, sneezing, vomiting) to utilize the same neuron without being attenuated by the GABAergic modulation. PMID- 12574465 TI - Binding sites, singly bound states, and conformation coupling shape GABA-evoked currents. AB - The time course of GABA-evoked currents is the main source of information on the GABA(A) receptor gating. Since the kinetics of these currents depends on the transitions between several receptor conformations, it is a major challenge to define the relations between current kinetics and the respective rate constants of the microscopic gating scheme. The aim of this study was to further explore the impact of different GABA(A) receptor conformations on the kinetics of currents elicited by ultra-fast GABA applications. We show that the rising phase and amplitude of GABA-evoked currents depend on desensitization and singly bound states. The occupancy of bound receptors depends not only on binding properties but also on opening/closing and desensitization. The impact of such functional coupling between channel states is critical in conditions of high non-equilibrium typical for synaptic transmission. The concentration dependence of the rising phase of the GABA-elicited current indicates positive cooperativity between agonist binding sites. We provide evidence that preequilibration at low GABA concentrations reduce GABA-evoked currents due to receptor trapping in a singly bound desensitized state. PMID- 12574466 TI - Cerebellar control of constrained and unconstrained movements. I. Nuclear inactivation. AB - The aim of this study was to determine in monkeys if inactivation of dentate and lateral interposed deep cerebellar nuclei preferentially impairs certain movements relative to others. Constrained movements of the digits were trained with digits, hand, and elbow constrained in a cast. Simple movements were flexion of Thumb or Index. A compound movement was simultaneous flexion of Thumb+Index. An unconstrained movement consisted of a reach to, pinch of, and retrieval of a small food reward (Reach+Pinch). In two monkeys we mapped the dentate and interpositus with 66 injections of muscimol (3 microl of 5 microg/microl). Thirty two percent of the injections resulted in increased reaction times of Thumb, Index, and Thumb+Index (mean = 24, 24, 28 + 26, respectively). Fifty percent of the injections impaired Reach+Pinch, producing target overshoot, curved reach trajectory, missed target (X and Y errors), and clumsy pinch with dropped fruit bits. Inactivation impaired each and all of Thumb, Index, Thumb+Index, and Reach+Pinch in 27%, only Reach+Pinch in 23%, and only Thumb, Index, Thumb+Index in 5% of injections. In sum, all types of movement were impaired. Thumb+Index was no more impaired than Thumb or Index alone, suggesting that the lateral cerebellar nuclei are not specifically required for combining movements of the two digits when constrained. Reach+Pinch appeared so greatly impaired and Thumb, Index, Thumb+Index so little as to be consistent with the hypothesis that a principal role of the cerebellum is to control those interactions that occur between body segments in natural unconstrained movements. However, the fact that all movements were impaired shows that the cerebellum contributes to the control of all movements. PMID- 12574467 TI - Cerebellar control of constrained and unconstrained movements. II. EMG and nuclear activity. AB - The aim of this study was to see in monkeys if neurons in dentate and lateral interposed deep cerebellar nuclei are preferentially active in relation to certain movements relative to others. Simple and compound digit movements were trained with digits, hand, and elbow constrained in a cast. The constrained simple movement was flexion of Thumb or Index; the constrained compound movement, flexion of Thumb+Index. An unconstrained compound movement consisted of a reach to, pinch of, and retrieval of a small food reward (Reach+Pinch). Electromyographic (EMG) recording showed that many muscles in the upper extremity, shoulder girdle, and trunk were active in all movements. EMG/muscle stimulation during the constrained digit movements showed that the digit prime movers were active during, sufficient for, and necessary for performance of these digit tasks. By contrast, EMG/muscle stimulation showed that the proximal muscles (though co-active during the tasks) were neither sufficient nor necessary for performance of the digit tasks. A fraction of those neurons that were active during both the constrained and the unconstrained movements fired at a higher frequency during the unconstrained task. Some neurons were active during Reach+Pinch only; a few others were active during one or more of Thumb, Index, Thumb+Index only. There was no distinct preferential discharge relationship to the compound Thumb+Index as compared with the simple Thumb or Index. These correlational data are consistent with an interpretation that the cerebellar discharge influenced all of these movements-simple and compound, constrained, and unconstrained-no one type seemingly more than any other. PMID- 12574468 TI - Fast rhythmic bursting can be induced in layer 2/3 cortical neurons by enhancing persistent Na+ conductance or by blocking BK channels. AB - Fast rhythmic bursting (or "chattering") is a firing pattern exhibited by selected neocortical neurons in cats in vivo and in slices of adult ferret and cat brain. Fast rhythmic bursting (FRB) has been recorded in certain superficial and deep principal neurons and in aspiny presumed local circuit neurons; it can be evoked by depolarizing currents or by sensory stimulation and has been proposed to depend on a persistent g(Na) that causes spike depolarizing afterpotentials. We constructed a multicompartment 11-conductance model of a layer 2/3 pyramidal neuron, containing apical dendritic calcium-mediated electrogenesis; the model can switch between rhythmic spiking (RS) and FRB modes of firing, with various parameter changes. FRB in this model is favored by enhancing persistent g(Na) and also by measures that reduce [Ca(2+)](i) or that reduce the conductance of g(K(C)) (a fast voltage- and Ca(2+)-dependent conductance). Axonal excitability plays a critical role in generating fast bursts in the model. In vitro experiments in rat layer 2/3 neurons confirmed (as shown previously by others) that RS firing could be switched to fast rhythmic bursting, either by buffering [Ca(2+)](i) or by enhancing persistent g(Na). In addition, our experiments confirmed the model prediction that reducing g(KC) (with iberiotoxin) would favor FRB. During the bursts, fast prepotentials (spikelets) could occur that did not originate in apical dendrites and that appear to derive from the axon. We suggest that modulator-induced regulation of [Ca(2+)] dynamics or of BK channel conductance, for example via protein kinase A, could play a role in determining the firing pattern of neocortical neurons; specifically, such modulation could play a role in regulating whether neurons respond to strong stimulation with fast rhythmic bursts. PMID- 12574469 TI - Neural activity in primary motor and dorsal premotor cortex in reaching tasks with the contralateral versus ipsilateral arm. AB - To investigate the effector dependence of task-related neural activity in dorsal premotor (PMd) and primary motor cortex (M1), directional tuning functions were compared between instructed-delay reaching tasks performed separately with either the contralateral or the ipsilateral limb. During presentation of the instructional cue, the majority (55/90, 61%) of cells in PMd were tuned with both arms, and their dynamic range showed a trend for stronger discharge with the contralateral arm. Most strikingly, however, the preferred direction of most of these latter cells (41/55, 75%) was not significantly different between arms. During movement, many PMd cells continued to be tuned with both arms (53/90, 59%), with a trend for increasing directional differences between the arms over the course of the trial. In contrast, during presentation of the instructional cue only 5/74 (7%) cells in M1 were tuned with both arms. During movement, about half of M1 cells (41/74, 55%) were tuned with both arms but the preferred directions of their tuning functions were often very different and there was a strong bias toward greater discharge rates when the contralateral arm was used. Similar trends were observed for EMG activity. In conclusion, M1 is strongly activated during movements of the contralateral arm, but activity during ipsilateral arm movements is also common and usually different from that seen with the contralateral arm. In contrast, a major component of task-related activity in PMd represents movement in a more abstract or task-dependent and effector-independent manner, especially during the instructed-delay period. PMID- 12574470 TI - Distinct local circuits between neocortical pyramidal cells and fast-spiking interneurons in young adult rats. AB - Connections between layer V pyramidal cells and GABAergic fast-spiking interneurons (pyramidal-FS) were studied by paired recordings combined with morphological analyses in acute neocortical slices from 28- to 52-day-old rats. Pairs of spikes elicited in pyramidal cells at a stimulation rate of 0.2 Hz induced unitary excitatory postsynaptic currents (EPSCs) in FS interneurons that displayed facilitation (48%), depression (38.5%), or neither depression nor facilitation (13.5%). Analyses of the EPSC amplitude distributions indicate that depressing connections always showed multiple functional release sites. On the contrary, facilitating connections consisted either of one or several release sites. At a holding potential of -72 mV, the quantal size (q) and the release probability (p) of facilitating connections with a single release site were -21.9 +/- 7.5 pA and 0.49 +/- 0.19 (SD), respectively. The mean q and the estimated number of release sites (n) at connections showing multiple sites were obtained by decreasing the release probability and did not differ between depressing and facilitating synapses (depressing connections: q = -15.3 +/- 2.5 pA, n = 5.1 +/- 3, facilitating connections: q = -23.9 +/- 9.8 pA, n = 7.8 +/- 5.4). However, the quantal content at facilitating synapses with multiple sites (1.9 +/- 1.5) was significantly different from that at depressing connections (4.1 +/- 3.9). Finally, quantitative morphological analyses revealed that most of the pyramidal cells displaying facilitation can be differentiated from those displaying depression by a more densely branched apical dendritic tree. Therefore two types of morphologically distinct pyramidal cells form excitatory connections with FS interneurons that differ in their short-term plasticity characteristics. Facilitating and depressing connections may provide a differential control of the temporal information processing of FS cells and thus finely regulate the inhibitory effect of these interneurons in neocortical networks of young adult rats. PMID- 12574471 TI - 5-HT2 receptors promote plateau potentials in turtle spinal motoneurons by facilitating an L-type calcium current. AB - The effects of serotonin (5-HT) on intrinsic properties of spinal motoneurons were investigated with intracellular recordings in a slice preparation from adult turtles. In 55% of the cells that were recorded, addition of 5-HT to the extracellular medium promoted plateau potentials as revealed by the response to depolarizing current pulses applied through the intracellular electrode. In the remaining 45% of cells, 5-HT had an inhibitory effect. However, when tested with an applied electric field that preferentially polarizes distal dendrites, 5-HT facilitated plateau potentials in 100% of the cells. Plateau potentials were also promoted by 5-HT focally applied on a dendrite by iontophoresis. Applied near the soma, 5-HT either promoted plateau potentials or inhibited spike generation. The latter effect was accompanied by a decrease in input resistance. Voltage-clamp recordings showed that the facilitation of plateau potentials mediated by L-type Ca(2+) channels was due to activation of 5-HT(2) receptors. These findings show that 5-HT regulates intrinsic properties of motoneurons in opposite ways: activation of 5-HT receptors in the soma region inhibits spike generation and plateau potentials, while activation of 5-HT(2) receptors in the dendrites and the soma region promotes spiking by facilitation of plateau potentials mediated by L-type Ca(2+) channels. PMID- 12574472 TI - Functional coupling of motor units is modulated during walking in human subjects. AB - Time- and frequency-domain analysis of the coupling between pairs of electromyograms (EMG) recorded from leg muscles was investigated during walking in healthy human subjects. For two independent surface EMG signals from the tibialis anterior (TA) muscle, coupling estimated from coherence measurements was observed at frequencies /=70% of the current was mediated by D,L threo-beta-hydroxyaspartate (THA)-sensitive Glu transporters. Glu inward currents were completely and reversibly inhibited by depolarization of Bergmann glial cells to positive membrane potentials allowing biophysical inhibition of Glu uptake into a single glial cell. Inhibition of Glu transport into Bergmann glial cells by voltage-clamping the cell to depolarized potentials caused a reversible increase in spontaneous EPSC frequency in the Purkinje cell. This increase could also be achieved by pharmacological inhibition of Glu transport with the Glu transport inhibitor THA, suggesting that inhibition of Glu uptake into Bergmann glial cells is responsible for the modulation of postsynaptic EPSCs. THA modulation of spontaneous EPSCs could only be observed in the absence of TTX, suggesting primarily a presynaptic effect. Taken together these data suggest that glial Glu uptake can profoundly affect excitatory transmission in the cerebellum, most likely by regulating presynaptic glutamate release. PMID- 12574475 TI - Functional properties of brain areas associated with motor execution and imagery. AB - Imagining motor acts is a cognitive task that engages parts of the executive motor system. While motor imagery has been intensively studied using neuroimaging techniques, most studies lack behavioral observations. Here, we used functional MRI to compare the functional neuroanatomy of motor execution and imagery using a task that objectively assesses imagery performance. With surface electromyographic monitoring within a scanner, 10 healthy subjects performed sequential finger-tapping movements according to visually presented number stimuli in either a movement or an imagery mode of performance. We also examined effects of varied and fixed stimulus types that differ in stimulus dependency of the task. Statistical parametric mapping revealed movement-predominant activity, imagery-predominant activity, and activity common to both movement and imagery modes of performance (movement-and-imagery activity). The movement-predominant activity included the primary sensory and motor areas, parietal operculum, and anterior cerebellum that had little imagery-related activity (-0.1 ~ 0.1%), and the caudal premotor areas and area 5 that had mild-to-moderate imagery-related activity (0.2 ~ 0.7%). Many frontoparietal areas and posterior cerebellum demonstrated movement-and-imagery activity. Imagery-predominant areas included the precentral sulcus at the level of middle frontal gyrus and the posterior superior parietal cortex/precuneus. Moreover, activity of the superior precentral sulcus and intraparietal sulcus areas, predominantly on the left, was associated with accuracy of the imagery task performance. Activity of the inferior precentral sulcus (area 6/44) showed stimulus-type effect particularly for the imagery mode. A time-course analysis of activity suggested a functional gradient, which was characterized by a more "executive" or more "imaginative" property in many areas related to movement and/or imagery. The results from the present study provide new insights into the functional neuroanatomy of motor imagery, including the effects of imagery performance and stimulus-dependency on brain activity. PMID- 12574477 TI - Preparatory set associated with pro-saccades and anti-saccades in humans investigated with event-related FMRI. AB - Previous studies have shown that the BOLD functional MRI (fMRI) signal is increased in several cortical areas when subjects perform anti-saccades compared with pro-saccades. It remains unknown, however, whether this increase is due to an increased cortical motor signal for anti-saccades or due to differences in preparatory set between pro- and anti-saccade trials. To address this question, we measured event-related fMRI in a paradigm that allowed us to separate instruction-related brain activity from saccade-related brain activity. In this paradigm, the instruction to either generate a pro-saccade or an anti-saccade was conveyed by a switch in the color of the central fixation stimulus and preceded the presentation of a peripheral stimulus by either 6, 10, or 14 s. Cortical areas were functionally mapped using the general linear model comparing standard pro- and anti-saccade blocks with fixation blocks. When the trials were aligned on the onset of the instruction stimulus, bilateral frontal eye fields and right hemisphere dorsolateral prefrontal cortex showed an increased signal during the instruction period on anti-saccade trials as compared with pro-saccade trials. When the trials were aligned on the movement stimulus and the instruction period activity was subtracted, there were no differences between pro- and anti saccades. This finding suggests that the increased cortical activation found in previous blocked designs originates predominately from differences in preparatory set and not from differences in the motor signal between pro- and anti-saccades. PMID- 12574476 TI - Receptive fields and response properties of neurons in layer 4 of ferret visual cortex. AB - The ferret has become a model animal for studies exploring the development of the visual system. However, little is known about the receptive-field structure and response properties of neurons in the adult visual cortex of the ferret. We performed single-unit recordings from neurons in layer 4 of adult ferret primary visual cortex to determine the receptive-field structure and visual-response properties of individual neurons. In particular, we asked what is the spatiotemporal structure of receptive fields of layer 4 neurons and what is the orientation selectivity of layer 4 neurons? Receptive fields of layer 4 neurons were mapped using a white-noise stimulus; orientation selectivity was determined using drifting, sine-wave gratings. Our results show that most neurons (84%) within layer 4 are simple cells with elongated, spatially segregated, ON and OFF subregions. These neurons are also selective for stimulus orientation; peaks in orientation-tuning curves have, on average, a half-width at half-maximum response of 21.5 +/- 1.2 degrees (mean +/- SD). The remaining neurons in layer 4 (16%) lack orientation selectivity and have center/surround receptive fields. Although the organization of geniculate inputs to layer 4 differs substantially between ferret and cat, our results demonstrate that, like in the cat, most neurons in ferret layer 4 are orientation-selective simple cells. PMID- 12574478 TI - Directional sensitivity of neurons in the primary auditory (AI) cortex: effects of sound-source intensity level. AB - Transient sounds were delivered from different directions in virtual acoustic space while recording from single neurons in primary auditory cortex (AI) of cats under general anesthesia. The intensity level of the sound source was varied parametrically to determine the operating characteristics of the spatial receptive field. The spatial receptive field was constructed from the onset latency of the response to a sound at each sampled direction. Spatial gradients of response latency composing a receptive field are due partially to a systematic co-dependence on sound-source direction and intensity level. Typically, at any given intensity level, the distribution of response latency within the receptive field was unimodal with a range of approximately 3-4 ms, although for some cells and some levels, the spread could be as much as 20 or as little as 2 ms. Response latency, averaged across directions, differed among neurons for the same intensity level, and also differed among intensity levels for the same neuron. Generally, increases in intensity level resulted in decreases in the mean and variance, which follows an inverse Gaussian distribution. Receptive field models, based on response latency, are developed using multiple parameters (azimuth, elevation, intensity), validated with Monte Carlo simulation, and their spatial filtering described using spherical harmonic analysis. Observations from an ensemble of modeled receptive fields are obtained by linking the inverse Gaussian density to the probabilistic inverse problem of estimating sound-source direction and intensity. Upper bounds on acuity is derived from the ensemble using Fisher information, and the predicted patterns of estimation errors are related to psychophysical performance. PMID- 12574479 TI - Activity in the supplementary motor area related to learning and performance during a sequential visuomotor task. AB - Monkeys were trained in a serial reaction time task to produce hand movements according to changing locations of visual targets. In most trials, targets followed the same sequence repeatedly, whereas in other trials targets were presented in random locations or switched unpredictably between two alternative sequences. Single-unit activity was recorded from the caudal supplementary motor area (SMA-proper). Based on the activity associated with random movement sequences, effects of hand position and movement direction were evaluated. Activity was influenced by the hand position in ~60% of the neurons, and the movement direction influenced the activity of 51% of the neurons. In addition, 37 and 71% of SMA neurons displayed nonstationarity in their activity across successive movements within a given trial and across trials, respectively. Such nonstationarity in the ongoing neural activity and the effects of performance related variables were evaluated using a regression model and separated from learning-related activity changes. About a third of SMA neurons displayed gradual changes in neural activity related to experience with a movement sequence across trials. Furthermore, about a quarter of SMA neurons showed similar changes within individual trials. When the individual movements included in the frequently repeated movement sequences were introduced unexpectedly, learning-related changes in neural activity were reduced, indicating that many SMA neurons changed their activity in relation to the learning of particular movement sequences. These results suggest that the pattern of neural activity in the cortical network involved in the control of movement sequences can be modified continuously by experience. PMID- 12574480 TI - Rhythmically discharging basal forebrain units comprise cholinergic, GABAergic, and putative glutamatergic cells. AB - The basal forebrain plays important roles in arousal, learning, and memory by stimulating cortical activation characterized by rhythmic slow theta and high frequency beta-gamma activities. Although cholinergic neurons play a significant part in these roles, other, including GABAergic, neurons appear to contribute. Using juxtacellular labeling with neurobiotin of neurons recorded within the magnocellular preoptic-substantia innominata area in urethan-anesthetized rats, we show that in addition to cells that are cholinergic or GABAergic, other cells that are neither fire rhythmically in correlation with stimulation-induced rhythmic slow activity on the cortex. Neurons with the characteristics of the noncholinergic/nonGABAergic cells contain phosphate-activated glutaminase (PAG), the synthetic enzyme for transmitter glutamate and may thus be glutamatergic. Within their oscillatory spike trains, putative glutamatergic neurons fire at a lower frequency (~20 Hz) than the GABAergic neurons (~40 Hz) and the cholinergic neurons (average: 75 Hz), whose spike trains include high-frequency bursts. The three groups all discharge rhythmically at a slow frequency in correlation with rhythmic slow activity recorded on the prefrontal, entorhinal, piriform and olfactory bulb cortices. The predominant slow frequency corresponds to the respiratory-olfactory rhythm, which is commonly slower than, yet can be as fast as, the hippocampal theta rhythm during certain coordinated behaviors, such as sniffing-whisking. While stimulating higher frequency beta-gamma activities, putative glutamatergic together with GABAergic and cholinergic cells may thus collectively modulate rhythmic slow activity and thereby promote coherent processing and plasticity across distributed cortical networks during coordinated behaviors and states. PMID- 12574481 TI - Dissociation of spatial-, object-, and sound-coding neurons in the mediodorsal nucleus of the primate thalamus. AB - The mediodorsal nucleus (MD) is the thalamic gateway to the prefrontal cortex, an area of the brain associated with spatial and object working memory functions. We have recorded single-neuron activities from the MD nucleus in monkeys trained to perform spatial tasks with peripheral visual stimuli and a nonspatial task with foveally presented pictures of objects and faces-tasks identical to those we have previously used to map regional specializations in the dorso- and ventro-lateral prefrontal cortex, respectively. We found that MD neurons exhibited categorical specificity-either responding selectively to locations in the spatial tasks or preferentially to specific representations of faces and objects in the nonspatial task. Spatially tuned neurons were located in parts of the MD connected with the dorsolateral prefrontal cortex while neurons responding to the identity of stimuli mainly occupied more ventral positions in the nucleus that has its connections with the inferior prefrontal convexity. Neuronal responses to auditory stimuli were also examined, and vocalization sensitive neurons were found in more posterior portions of the MD. We conclude that MD neurons are dissociable by their spatial and nonspatial coding properties in line with their cortical connections and that the principle of information segregation in cortico cortical pathways extends to the "association" nuclei of the thalamus. PMID- 12574482 TI - Auditory influences on visual temporal rate perception. AB - Visual stimuli are known to influence the perception of auditory stimuli in spatial tasks, giving rise to the ventriloquism effect. These influences can persist in the absence of visual input following a period of exposure to spatially disparate auditory and visual stimuli, a phenomenon termed the ventriloquism aftereffect. It has been speculated that the visual dominance over audition in spatial tasks is due to the superior spatial acuity of vision compared with audition. If that is the case, then the auditory system should dominate visual perception in a manner analogous to the ventriloquism effect and aftereffect if one uses a task in which the auditory system has superior acuity. To test this prediction, the interactions of visual and auditory stimuli were measured in a temporally based task in normal human subjects. The results show that the auditory system has a pronounced influence on visual temporal rate perception. This influence was independent of the spatial location, spectral bandwidth, and intensity of the auditory stimulus. The influence was, however, strongly dependent on the disparity in temporal rate between the two stimulus modalities. Further, aftereffects were observed following approximately 20 min of exposure to temporally disparate auditory and visual stimuli. These results show that the auditory system can strongly influence visual perception and are consistent with the idea that bimodal sensory conflicts are dominated by the sensory system with the greater acuity for the stimulus parameter being discriminated. PMID- 12574483 TI - Coding of horizontal disparity and velocity by MT neurons in the alert macaque. AB - We performed the first large-scale (n = 501), quantitative study of horizontal disparity tuning in the middle temporal (MT) visual area of alert, fixating macaque monkeys. Using random-dot stereograms, we quantified the direction tuning, speed tuning, horizontal disparity tuning, and size tuning of each neuron. The vast majority (93%) of MT neurons were significantly tuned for horizontal disparity. Although disparity tuning was generally quite robust, the average disparity sensitivity of MT neurons was significantly weaker than their direction or speed sensitivity as quantified using both an index of response modulation and an index of signal-to-noise ratio. Disparity tuning was not correlated with direction or size tuning but tended to be broader and weaker for neurons that preferred faster speeds of motion. By comparison with recent studies, we find that disparity selectivity in MT is substantially stronger than that seen in either primary visual cortex (V1) or area V4. In addition, MT neurons are more broadly tuned for disparity than V1 neurons at comparable eccentricities. Disparity tuning curves are very well described by Gabor functions for >80% of MT neurons. The distribution of Gabor phases shows clear bimodality, indicating that MT neurons tend to have odd-symmetric disparity tuning (unlike neurons in V1). The preferred disparities were more strongly correlated with the phase parameter of the Gabor function than with the positional offset parameter. In fact, for neurons with preferred disparities close to zero, the positional offset tended to oppose the phase shift in specifying the disparity preference. We suggest that this result reflects a strategy used to finely distribute the disparity preferences of MT neurons, given the predominance of odd-symmetry and broad tuning. PMID- 12574484 TI - Functional organization of the cat visual cortex in relation to the representation of a uniform surface. AB - Neuronal activity in the early visual cortex has been extensively studied from the standpoint of contour representation. On the other hand, representation of the interior of a surface surrounded by a contour is much less well understood. Several studies have identified neurons activated by a uniform surface covering their receptive fields, but their distribution within the cortex is not yet known. The aim of the present study was to obtain a better understanding of the distribution of such neurons in the visual cortex. Using optical imaging of intrinsic signals, we found that there are a group of surface-responsive regions located in area 18, along the area 17/18 border, that tend to overlap the singular points of the orientation-preference map. Extracellular recordings confirmed that neurons responsive to uniform plane stimuli are accumulated in these regions. Such neurons also existed outside the surface-responsive regions around the singular points. These results suggest that there exists a functional organization related to the representation of a uniform surface in the early visual cortex. PMID- 12574485 TI - Neural networks for the coordination of the hands in time. AB - Without practice, bimanual movements can typically be performed either in phase or in antiphase. Complex temporal coordination, e.g., during movements at different frequencies with a noninteger ratio (polyrhythms), requires training. Here, we investigate the organization of the neural control systems for in-phase, antiphase, and polyrhythmic coordination using functional magnetic resonance imaging (fMRI). Brisk rhythmic tapping with the index fingers was used as a model behavior. We demonstrate different patterns of brain activity during in-phase and antiphase coordination. In-phase coordination was characterized by activation of the right anterior cerebellum and cingulate motor area (CMA). Antiphase coordination was accompanied by extensive fronto-parieto-temporal activations, including the supplementary motor area (SMA), the preSMA, and the bilateral inferior parietal gyri, premotor cortex, and superior temporal gyri. When contrasting polyrhythmic tapping with in-phase tapping, activity was seen in the same set of brain regions, and in the posterior cerebellum and the CMA. Antiphase and polyrhythmic coordination may thus to a large extent use common neural control circuitry. In a separate experiment, we analyzed the neural control of the rhythmic structure and the serial order of finger movements during polyrhythmic tapping. Polyrhythmic tapping was compared with an isochronous coordination pattern that retained the same serial order of finger movements as the polyrhythm. This experiment showed that the preSMA and the bilateral superior temporal gyri may be crucial for the rhythmic control of polyrhythmic tapping, while the cerebellum, the CMA, and the premotor cortices presumably are more involved in the ordinal control of the sequence of finger movements. PMID- 12574486 TI - Dynamical organization of directional tuning in the primate premotor and primary motor cortex. AB - Although previous studies have shown that activity of neurons in the motor cortex is related to various movement parameters, including the direction of movement, the spatial pattern by which these parameters are represented is still unresolved. The current work was designed to study the pattern of representation of the preferred direction (PD) of hand movement over the cortical surface. By studying pairwise PD differences, and by applying a novel implementation of the circular variance during preparation and movement periods in the context of a center-out task, we demonstrate a nonrandom distribution of PDs over the premotor and motor cortical surface of two monkeys. Our analysis shows that, whereas PDs of units recorded by nonadjacent electrodes are not more similar than expected by chance, PDs of units recorded by adjacent electrodes are. PDs of units recorded by a single electrode display the greatest similarity. Comparison of PD distributions during preparation and movement reveals that PDs of nearby units tend to be more similar during the preparation period. However, even for pairs of units recorded by a single electrode, the mean PD difference is typically large (45 degrees and 75 degrees during preparation and movement, respectively), so that a strictly modular representation of hand movement direction over the cortical surface is not supported by our data. PMID- 12574487 TI - Functional interaction of auxiliary subunits and synaptic proteins with Ca(v)1.3 may impart hair cell Ca2+ current properties. AB - We assessed the functional determinants of the properties of L-type Ca(2+) currents in hair cells by co-expressing the pore-forming Ca(V)1.3alpha(1) subunit with the auxiliary subunits beta(1A) and/or alpha(2delta). Because Ca(2+) channels in hair cells are poised to interact with synaptic proteins, we also co expressed the Ca(V)1.3alpha(1) subunit with syntaxin, vesicle-associated membrane protein (VAMP), and synaptosome associated protein of 25 kDa (SNAP25). Expression of the Ca(V)1.3alpha(1) subunit in human embryonic kidney cells (HEK 293) produced a dihydropyridine (DHP)-sensitive Ca(2+) current (peak current density 2.0 +/- 0.2 pA/pF; n = 11). Co-expression with beta(1A) and alpha(2delta) subunits enhanced the magnitude of the current (peak current density: Ca(V)1.3alpha(1) + beta(1A) = -4.3 +/- 0.8 pA/pF, n = 10; Ca(V)1.3alpha(1) + beta(1A) + alpha(2delta) = -4.1 +/- 0.6 pA/pF, n = 9) and produced a leftward shift of approximately 9 mV in the voltage-dependent activation of the currents. Furthermore, co-expression of Ca(V)1.3alpha(1) with syntaxin/VAMP/SNAP resulted in at least a twofold increase in the peak current density (-4.7 +/- 0.2 pA/pF; n = 11) and reduced the extent of inactivation of the Ca(2+) currents. Botulinum toxin, an inhibitor of syntaxin, accelerated the inactivation profile of Ca(2+) currents in hair cells. Immunocytochemical data also indicated that the Ca(2+) channels and syntaxin are co-localized in hair cells, suggesting there is functional interaction of the Ca(V)1.3alpha(1) with auxiliary subunits and synaptic proteins, that may contribute to the distinct properties of the DHP sensitive channels in hair cells. PMID- 12574488 TI - Effects of high-frequency stimulation in the internal globus pallidus on the activity of thalamic neurons in the awake monkey. AB - The reduction in symptoms of Parkinson's disease produced by high-frequency stimulation (HFS) in the internal globus pallidus (GPi) has been proposed to be due to stimulus-induced inactivation of pallidal neurons and resulting disinhibition of thalamic neurons. We tested this in awake Macaca fascicularis by stimulating between pairs of electrodes inserted into GPi under electrophysiological control and recording the responses evoked in thalamic neurons. HFS produced a reduction, not an increase, in discharge frequency during the stimulus train in 77% of the responsive thalamic neurons. Only 16% of the responsive cells showed an increase in discharge during stimulation and, for some of these, stimulation at a similar intensity produced contralateral muscle contraction, a probable sign of current spread to the internal capsule. The few thalamic neurons studied during bursting had a reduction in burst frequency and duration during HFS. We conclude that high-frequency stimulation within GPi does not necessarily facilitate thalamic discharge, and it may act, instead, to interrupt abnormal patterns of thalamic discharge associated with parkinsonian symptoms. PMID- 12574489 TI - The anterior cingulate and reward-guided selection of actions. AB - Macaques were taught a reward-conditional response selection task; they learned to associate each of two different actions to each of two different rewards and to select actions that were appropriate for particular rewards. They were also taught a visual discrimination learning task. Cingulate lesions significantly impaired selection of responses associated with different rewards but did not interfere with visual discrimination learning or performance. The results suggest that 1) the cingulate cortex is concerned with action reward associations and not limited to just detecting when actions lead to errors and 2) that the cingulate cortex's function is limited to action reinforcer associations and it is not concerned with stimulus reward associations. PMID- 12574490 TI - Context-dependent anticipation of different task dynamics: rapid recall of appropriate motor skills using visual cues. AB - Recent studies have reported that human subjects show varying degrees of ability to use contextual cues to recall the motor skills required to compensate for different dynamic external force fields during arm movements. In particular, the subjects showed little or no ability to use color cues to adjust motor outputs in anticipation of the perturbing fields after limited periods of training that were sufficient to learn to compensate for the fields themselves. This is unexpected since humans and monkeys can use color cues to perform a wide range of visuomotor tasks. Here we show that a monkey with extensive practice compensating for viscous fields in an elbow-movement task can use associated color cues to adjust motor output in anticipation of an impending field before physically encountering it. PMID- 12574491 TI - Photoelectron angular distributions. AB - Angle-resolved photoelectron spectroscopy has been performed for more than 70 years in various guises, but recently its potential to help solve in detail problems in the photoionization dynamics and intramolecular dynamics of gas-phase molecules has been recognized. One key development has been the design of experiments in appropriate geometries to extract information that pertains to the molecular frame, another has been the development of imaging spectrometers, and a third is the use of ultrafast lasers to cause photoionization. In this review, which is aimed at experimentalists, simple expressions for photoelectron angular distributions (PADs) in various experimental geometries are given and their applications explained. PMID- 12574492 TI - Master equation models for chemical reactions of importance in combustion. AB - The master equation provides a quantitative description of the interaction between collisional energy transfer and chemical reaction for dissociation, isomerization, and association processes. The approach is outlined for both irreversible and reversible dissociation, isomerization, and association reactions. There is increasing interest, especially in combustion, in association reactions that involve several linked potential wells, with the possibility of isomerization, collisional stabilization, and dissociation along several product channels. A major aim of the application of the master equation to such systems is the linking of the eigenvalues obtained by its solution to the rate coefficients for the phenomenological chemical reactions that describe the system and that are used in combustion models. The approach is illustrated by reference to the reactions C2H5 + O2, H + SO2, and the dissociation and isomerization of alkyl radicals. PMID- 12574493 TI - Optical diagnostics for thin film processing. AB - Optical diagnostics are used to probe the plasma or neutral gas above the substrate, particles in the gas or on the surface, the film surface and reactor walls, the film itself, and the substrate during thin film processing. The development and application of optical probes are highlighted, in particular for analyzing plasma/gas phase intermediates and products and film composition, and performing metrology, thermometry, and endpoint detection and control. Probing etching (particularly plasma etching) and deposition (particularly epitaxy) are emphasized. PMID- 12574494 TI - Color vision. AB - Color vision starts with the absorption of light in the retinal cone photoreceptors, which transduce electromagnetic energy into electrical voltages. These voltages are transformed into action potentials by a complicated network of cells in the retina. The information is sent to the visual cortex via the lateral geniculate nucleus (LGN) in three separate color-opponent channels that have been characterized psychophysically, physiologically, and computationally. The properties of cells in the retina and LGN account for a surprisingly large body of psychophysical literature. This suggests that several fundamental computations involved in color perception occur at early levels of processing. In the cortex, information from the three retino-geniculate channels is combined to enable perception of a large variety of different hues. Furthermore, recent evidence suggests that color analysis and coding cannot be separated from the analysis and coding of other visual attributes such as form and motion. Though there are some brain areas that are more sensitive to color than others, color vision emerges through the combined activity of neurons in many different areas. PMID- 12574495 TI - Epidemiology of neurodegeneration. AB - Alzheimer's disease, Parkinson's disease, and motor neuron disease share a propensity to occur with increasing age and as either a sporadic or a familial disorder. A number of behavioral and environmental risk factors have been proposed for each disorder, but most associations lack consistency and specificity. Over the last decade the remarkable frequency of these disorders has become apparent, and the identification of mutations in genes has provided the means to understand their pathogenesis. Better and more accurate means to characterize and diagnose these diseases has greatly facilitated analytic epidemiology. The analysis of behavioral and genetic factors that may lower disease risk has led to clinical trials that are either in progress or being planned with the aim of preventing these disorders. PMID- 12574496 TI - Forces for morphogenesis investigated with laser microsurgery and quantitative modeling. AB - We investigated the forces that connect the genetic program of development to morphogenesis in Drosophila. We focused on dorsal closure, a powerful model system for development and wound healing. We found that the bulk of progress toward closure is driven by contractility in supracellular "purse strings" and in the amnioserosa, whereas adhesion-mediated zipping coordinates the forces produced by the purse strings and is essential only for the end stages. We applied quantitative modeling to show that these forces, generated in distinct cells, are coordinated in space and synchronized in time. Modeling of wild-type and mutant phenotypes is predictive; although closure in myospheroid mutants ultimately fails when the cell sheets rip themselves apart, our analysis indicates that beta(PS) integrin has an earlier, important role in zipping. PMID- 12574497 TI - Magma ascent and the pressurization of Mount Etna's volcanic system. AB - After a period of deflation during the 1991-1993 flank eruption, Mount Etna underwent a rapid inflation. Seismicity and ground deformation show that since 1994, a huge volume of magma intruded beneath the volcano, producing from 1998 onward a series of eruptions at the summit and on the flank of the volcano. The last of these, started on 27 October 2002, is still in progress and can be considered one of the most explosive eruptions of the volcano in recent times. Here we show how geodetic data and seismic deformation, between 1994 and 2001, indicate a radial compression around an axial intrusion, consistent with a repressurization of Mount Etna's plumbing system at a depth of 6 to 15 kilometers, which triggered most of the seismicity and provoked the dilatation of the volcano and the recent explosive eruptive activity. PMID- 12574498 TI - Gas-phase infrared spectrum of the protonated water dimer. AB - The protonated water dimer is a prototypical system for the study of proton transfer in aqueous solution. We report infrared photodissociation spectra of cooled H+(H2O)2 [and D+(D2O2] ions, measured between 620 and 1900 wave numbers (cm-1). The experiment directly probes the shared proton region of the potential energy surface and reveals three strong bands below 1600 cm-1 and one at 1740 cm 1 (for H5O2+). From a comparison to multidimensional quantum calculations, the three lower energy bands were assigned to stretching and bending fundamentals involving the O...H+...O moiety, and the highest energy band was assigned to a terminal water bend. These results highlight the importance of intermode coupling in shared proton systems. PMID- 12574499 TI - PUMA mediates the apoptotic response to p53 in colorectal cancer cells. AB - Although several genes that might mediate p53-induced apoptosis have been proposed, none have previously been shown to play an essential role in this process through a rigorous gene disruption approach. We used a gene-targeting approach to evaluate p53-mediated death in human colorectal cancer cells. Expression of p53 in these cells induces growth arrest through transcriptional activation of the cyclin-dependent kinase inhibitor p21. If p21 is disrupted via gene targeting, the cells die through apoptosis. If the PUMA gene is also disrupted in such cells, apoptosis is prevented. The effects of PUMA on apoptosis were observed after exogenous overexpression of p53 as well as after exposure to hypoxia, a physiologic activator of p53, and DNA damage. The PUMA protein interacts with Bcl-X(L) and promotes mitochondrial translocation and multimerization of Bax. Accordingly, genetic disruption of BAX makes cells resistant to the apoptosis resulting from PUMA expression. These results suggest that the balance between PUMA and p21 is pivotal in determining the responses to p53 activation and provide a model for understanding the basis of p53 mutations in human cancer. PMID- 12574500 TI - DNA unzipped under a constant force exhibits multiple metastable intermediates. AB - Single molecule studies, at constant force, of the separation of double-stranded DNA into two separated single strands may provide information relevant to the dynamics of DNA replication. At constant applied force, theory predicts that the unzipped length as a function of time is characterized by jumps during which the strands separate rapidly, followed by long pauses where the number of separated base pairs remains constant. Here, we report previously uncharacterized observations of this striking behavior carried out on a number of identical single molecules simultaneously. When several single lambda phage molecules are subject to the same applied force, the pause positions are reproducible in each. This reproducibility shows that the positions and durations of the pauses in unzipping provide a sequence-dependent molecular fingerprint. For small forces, the DNA remains in a partially unzipped state for at least several hours. For larger forces, the separation is still characterized by jumps and pauses, but the double-stranded DNA will completely unzip in less than 30 min. PMID- 12574501 TI - Inhibition of herpes simplex virus 1 gene expression by designer zinc-finger transcription factors. AB - The herpes simplex virus 1 (HSV-1) replicative cycle begins by binding of the viral activator, VP16, to a set of sequences in the immediate-early (IE) gene promoters. With the aim of inhibiting this cycle, we have constructed a number of synthetic zinc-finger DNA-binding peptides by using recently reported methods. Peptides containing either three or six fingers, targeted to a viral promoter, were engineered as fusions with a KOX-1 transcription repression domain. These proteins bound to the HSV-1 IE175k (ICP4) promoter, in vitro, with nanomolar or subnanomolar binding affinity. However, in a chloramphenicol acetyltransferase reporter system, only the six-finger protein was found to repress VP16-activated transcription significantly. Thus the longer array of zinc fingers is required to compete successfully against VP16, one of the most powerful natural activators known. We found that the HSV-1 replication cycle can be partially repressed by the six-finger peptide with the viral titer reduced by 90%. PMID- 12574502 TI - Repression of the HIV-1 5' LTR promoter and inhibition of HIV-1 replication by using engineered zinc-finger transcription factors. AB - Zinc finger domains are small DNA-binding modules that can be engineered to bind desired target sequences. Functional transcription factors can be made from these DNA-binding modules, by fusion with an appropriate effector domain. In this study, eight three-zinc-finger proteins (ZFPs) that bound HIV-1 sequences in vitro were engineered into transcription repressors by linking them to the Kruppel-associated box (KRAB) repressor domain (KOX1). One protein, ZFP HIVB-KOX, which bound to a 9-bp region overlapping two Sp1 sites, was found to repress a Tat-activated 5' LTR cellular HIV-reporter assay to almost basal levels. A related six-finger protein, HIVBA'-KOX, was made to target all three Sp1 sites in the 5' LTR promoter and efficiently inhibited both basal and Tat-activated transcription in unstimulated and mitogen-stimulated T cells. In contrast, a combination of two unlinked three-finger ZFPs, HIVA'-KOX and HIVB-KOX, which bind over the same region of DNA, resulted in less effective repression. Finally, HIVBA'-KOX was tested for its capacity to block viral replication in a cellular infection assay using the HIV-1 HXB2 strain. This ZFP was found to inhibit HIV-1 replication by 75% compared with control constructs, thus demonstrating the potential of this approach for antiviral therapy. PMID- 12574503 TI - Efficient T cell repertoire selection in tetraparental chimeric mice independent of thymic epithelial MHC. AB - Nonthymic epithelial cells were compared with thymic epithelial cells for their role in T cell repertoire selection. Tetraparental aggregation chimeras were generated from T and B cell-deficient mice (H-2(d) SCID or H-2(b) Rag-/-) and thymus-deficient nude mice (H-2(b) or H-2(d)). These tetraparental mice showed primary protective CD8(+) T cell responses, after lymphocytic choriomeningitis virus infection, that were peptide-specifically restricted to either thymic or nonthymic epithelial MHC at comparable levels. These chimeras also mounted neutralizing IgG responses dependent on cognate CD4(+) T helper cell activity restricted to nonthymic epithelial MHC. Therefore, in contrast to earlier results with irradiation or thymus chimeras, these relatively undisturbed tetraparental mice reveal that the MHC of nonthymic epithelial cells efficiently selects a functional T cell repertoire. PMID- 12574504 TI - Nanodissection and high-resolution imaging of the Rhodopseudomonas viridis photosynthetic core complex in native membranes by AFM. Atomic force microscopy. AB - In photosynthesis, highly organized multiprotein assemblies convert sunlight into biochemical energy with high efficiency. A challenge in structural biology is to analyze such supramolecular complexes in native membranes. Atomic force microscopy (AFM) with high lateral resolution, high signal-to-noise ratio, and the possibility to nanodissect biological samples is a unique tool to investigate multiprotein complexes at molecular resolution in situ. Here we present high resolution AFM of the photosynthetic core complex in native Rhodopseudomonas viridis membranes. Topographs at 10-A lateral and approximately 1-A vertical resolution reveal a single reaction center (RC) surrounded by a closed ellipsoid of 16 light-harvesting (LH1) subunits. Nanodissection of the tetraheme cytochrome (4Hcyt) subunit from the RC allows demonstration that the L and M subunits exhibit an asymmetric topography intimately associated to the LH1 subunits located at the short ellipsis axis. This architecture implies a distance distribution between the antenna and the RC compared with a centered location of the RC within a circular LH1, which may influence the energy transfer within the core complex. The LH1 subunits rearrange into a circle after removal of the RC from the core complex. PMID- 12574505 TI - Aging results in hypermethylation of ribosomal DNA in sperm and liver of male rats. AB - There is a concern that increased paternal age may be associated with altered fertility and an increased incidence of birth defects in man. In previous studies of aged male rats, we have found abnormalities in the fertility and in the embryos sired by older males. Aging in mammals is associated with alterations in the content and patterns of DNA methylation in somatic cells; however, little is known in regard to germ cells. A systematic search for global and gene-specific alterations of DNA methylation in germ cells and liver of male rats was done. Restriction landmark genomic scanning, a method used to determine specific methylation patterns of CpG island sequences, has revealed a region of the ribosomal DNA locus that is preferentially hypermethylated with age in both spermatozoa and liver. In contrast, all single copy CpG island sequences in spermatozoa and in liver remain unaltered with age. We further demonstrate that a large proportion of rat ribosomal DNA is normally methylated and that regional and site-specific differences exist in the patterns of methylation between spermatozoa and liver. We conclude that patterns of ribosomal DNA methylation in spermatozoa are vulnerable to the same age-dependent alterations that we observe in normal aging liver. Failure to maintain normal DNA methylation patterns in male germ cells could be one of the mechanisms underlying age-related abnormalities in fertility and progeny outcome. PMID- 12574506 TI - Designed FHIT alleles establish that Fhit-induced apoptosis in cancer cells is limited by substrate binding. AB - The FHIT gene is inactivated early in the development of many human tumors, and Fhit-deficient mice have increased cancer incidence. Viral reexpression of Fhit kills Fhit-deficient cells by induction of apoptosis. Fhit, a member of branch 2 of the histidine-triad superfamily of nucleoside monophosphate hydrolases and transferases, is a diadenosine polyphosphate hydrolase, the active-site histidine of which is not required for tumor suppression. To provide a rigorous test of the hypothesis that Fhit function depends on forming a complex with substrates, we designed a series of alleles of Fhit intended to reduce substrate-binding andor hydrolytic rates, characterized these mutants biochemically, and then performed quantitative cell-death assays on cancer cells virally infected with each allele. The allele series covered defects as great as 100,000-fold in k(cat) and increases as large as 30-fold in K(M). Nonetheless, when mutant FHIT genes were expressed in two human cancer cell lines containing FHIT deletions, reductions in apoptotic activity correlated exclusively with K(M). Mutants with 2- and 7-fold increases in K(M) significantly reduced apoptotic indices, whereas the mutant with a 30-fold increase in K(M) retained little cellular function. These data indicate that the proapoptotic function of Fhit is limited by substrate binding and is unrelated to substrate hydrolysis. PMID- 12574507 TI - Lysine-79 of histone H3 is hypomethylated at silenced loci in yeast and mammalian cells: a potential mechanism for position-effect variegation. AB - Methylation of lysine-79 (K79) within the globular domain of histone H3 by Dot1 methylase is important for transcriptional silencing and for association of the Sir silencing proteins in yeast. Here, we show that the level of H3-K79 methylation is low at all Sir-dependent silenced loci but not at other transcriptionally repressed regions. Hypomethylation of H3-K79 at the telomeric and silent mating-type loci, but not the ribosomal DNA, requires the Sir proteins. Overexpression of Sir3 concomitantly extends the domain of Sir protein association and H3-K79 hypomethylation at telomeres. In mammalian cells, H3-K79 methylation is found at loci that are active for V(D)J recombination, but not at recombinationally inactive loci that are heterochromatic. These results suggest that H3-K79 methylation is an evolutionarily conserved marker of active chromatin regions, and that silencing proteins block the ability of Dot1 to methylate histone H3. Further, they suggest that Sir proteins preferentially bind chromatin with hypomethylated H3-K79 and then block H3-K79 methylation. This positive feedback loop, and the reverse loop in which H3-K79 methylation weakens Sir protein association and leads to further methylation, suggests a model for position-effect variegation. PMID- 12574508 TI - The N-terminal domain of mammalian soluble epoxide hydrolase is a phosphatase. AB - The mammalian soluble epoxide hydrolase (sEH) is an enzyme with multiple functions, being implicated in detoxification of xenobiotic epoxides as well as in regulation of physiological processes such as blood pressure. The enzyme is a homodimer, in which each subunit is composed of two domains. The 35-kDa C terminal domain has an alpha/beta hydrolase fold and harbors the catalytic center for the EH activity. The 25-kDa N-terminal domain has a different alpha/beta fold and belongs to the haloacid dehalogenase superfamily of enzymes. The catalytic properties of the enzyme reported so far can all be explained by the action of the C-terminal domain alone. The function of the N-terminal domain, other than in structural stabilization of the dimer, has therefore remained unclear. By structural comparison of this domain to other haloacid dehalogenase family members, we identified a putative active site containing all necessary components for phosphatase activity. Subsequently, we found rat sEH hydrolyzed 4-nitrophenyl phosphate with a rate constant of 0.8 s(-1) and a K(m) of 0.24 mM. Recombinant human sEH lacking the C-terminal domain also displayed phosphatase activity. Presence of a phosphatase substrate did not affect epoxide turnover nor did epoxides affect dephosphorylation by the intact enzyme, indicating both catalytic sites act independently. The enzyme was unable to hydrolyze 4-nitrophenyl sulfate, suggesting its role in xenobiotic metabolism does not extend beyond phosphates. Thus, we propose this domain participates instead in the regulation of the physiological functions associated with sEH. PMID- 12574509 TI - Selective incorporation of influenza virus RNA segments into virions. AB - The genome of influenza A virus is comprised of eight viral RNA (vRNA) segments. Although the products of all eight vRNA segments must be present for viral replication, little is known about the mechanism(s) responsible for incorporation of these segments into virions. Two models have been proposed for the generation of infectious virions containing eight vRNA segments. The random-incorporation model assumes a common structural feature in all the vRNAs, enabling any combination of vRNAs to be incorporated randomly into virions. The selective incorporation model predicts the presence of specific structures in each vRNA segment, leading to the incorporation of a set of eight vRNA segments into virions. Here we demonstrate that eight different vRNA segments must be present for efficient virion formation and that sequences within the coding region of (and thus unique to) the neuraminidase vRNA possess a signal that drives incorporation of this segment into virions. These findings indicate a unique contribution from individual vRNA segments and thus suggest a selective (rather than random) mechanism of vRNA recruitment into virions. The neuraminidase vRNA incorporation signal and others yet to be identified should provide attractive targets for the attenuation of influenza viruses in vaccine production and the design of new antiviral drugs. PMID- 12574511 TI - Single-cell nonphotochemical hole burning of ovarian surface epithelial carcinoma and normal cells. AB - Persistent spectral nonphotochemical hole-burning (NPHB) spectroscopy has recently been applied to dye molecules in cells. The sensitivity of NPHB to the nanoenvironment of the probe is well established. It has been shown that NPHB applied to bulk suspensions of cultured human cells can distinguish between normal and cancer cells. Thus, NPHB has potential as a diagnostic cancer tool. For this reason, the methodology is referred to as hole-burning imaging, by analogy with MRI. The optical dephasing time (T(2)) of the dye in hole-burning image replaces the proton T(1) relaxation time in MRI. In addition to the T(2) mode of operation, there are four other modes including measurement of the spectral hole growth kinetics (HGK). Reported here is that the selectivity and sensitivity of NPHB operating in the HGK mode allow for distinction between normal and carcinoma cells at the single-cell level. The ovarian cell lines are ovarian surface epithelial cells with temperature-sensitive large T antigens (analogously normal) and ovarian surface epithelial carcinoma (OV167) cells. The mitochondrial specific dye used was rhodamine 800 (Molecular Probes). This carbocationic dye is highly specific for the outer and inner membranes of mitochondria. In line with the results for bulk suspensions of the two cell lines, the hole-burning efficiency for OV167 cells was found to be significantly higher than that for normal cells. Theoretical analysis of the HGK data leads to the conclusion that the degree of structural heterogeneity for the probe-host configurations in OV167 cells is lower than in the normal cells. Possible reasons for this are given. PMID- 12574510 TI - The soluble epoxide hydrolase encoded by EPXH2 is a bifunctional enzyme with novel lipid phosphate phosphatase activity. AB - The gene EPXH2 encodes for the soluble epoxide hydrolase (sEH), an enzyme involved in the regulation of cardiovascular and renal physiology containing two distinct domains connected via a proline-rich linker. The C-terminal domain containing the EH catalytic activity has been well studied. In contrast, a function for the N-terminal domain, which has high homology to the haloacid dehalogenase family of phosphatases, has not been definitively reported. In this study we describe the N-terminal domain as a functional phosphatase unaffected by a number of classic phosphatase inhibitors. Assuming a functional association between these catalytic activities, dihydroxy lipid phosphates were rationalized as potential endogenous substrates. A series of phosphorylated hydroxy lipids were therefore synthesized and found to be excellent substrates for the human sEH. The best substrate tested was the monophosphate of dihydroxy stearic acid (threo-910-phosphonoxy-hydroxy-octadecanoic acid) with K(m) = 21 +/- 0.3 microM, V(Max) = 338 +/- 12 nmol x min(-1) x mg(-1), and k(cat) = 0.35 +/- 0.01 s(-1). Therefore dihydroxy lipid phosphates are possible candidates for the endogenous substrates of the sEH N-terminal domain, which would represent a novel branch of fatty acid metabolism with potential signaling functions. PMID- 12574512 TI - Gray-scale photolithography using microfluidic photomasks. AB - The ability to produce three-dimensional (3D) microstructures is of increasing importance in the miniaturization of mechanical or fluidic devices, optical elements, self-assembling components, and tissue-engineering scaffolds, among others. Traditional photolithography, the most widely used process for microdevice fabrication, is ill-suited for 3D fabrication, because it is based on the illumination of a photosensitive layer through a "photomask" (a transparent plate that contains opaque, unalterable solid-state features), which inevitably results in features of uniform height. We have devised photomasks in which the light-absorbing features are made of fluids. Unlike in conventional photomasks, the opacity of the photomask features can be tailored to an arbitrary number of gray-scale levels, and their spatial pattern can be reconfigured in the time scale of seconds. Here we demonstrate the inexpensive fabrication of photoresist patterns that contain features of multiple and/or smoothly varying heights. For a given microfluidic photomask, the developed photoresist pattern can be predicted as a function of the dye concentrations and photomask dimensions. For selected applications, microfluidic photomasks offer a low-cost alternative to present gray-scale photolithography approaches. PMID- 12574513 TI - Assembly of core helices and rapid tertiary folding of a small bacterial group I ribozyme. AB - Compact but non-native intermediates have been implicated in the hierarchical folding of several large RNAs, but there is little information on their structure. In this article, ribonuclease and hydroxyl radical cleavage protection assays showed that base pairing of core helices stabilize a compact state of a small group I ribozyme from Azoarcus pre-tRNA(ile). Base pairing of the ribozyme core requires 10-fold less Mg(2+) than stable tertiary interactions, indicating that assembly of helices in the catalytic core represents a distinct phase that precedes the formation of native tertiary structure. Tertiary folding occurs in <100 ms at 37 degrees C. Such rapid folding is unprecedented among group I ribozymes and illustrates the association between structural complexity and folding time. A 3D model of the Azoarcus ribozyme was constructed by identifying homologous sequence motifs in rRNA. The model reveals distinct structural features, such as a large interface between the P4-P6 and P3-P9 domains, that may explain the unusual stability of the Azoarcus ribozyme and the cooperativity of folding. PMID- 12574514 TI - Dosage of Fgf8 determines whether cell survival is positively or negatively regulated in the developing forebrain. AB - FGF8 is known to be an important regulator of forebrain development. Here, we investigated the effects of varying the level of Fgf8 expression in the mouse forebrain. We detected two distinct responses, one that was proportionate with Fgf8 expression and another that was not. The latter response, which led to effects on cell survival, displayed a paradoxical relationship to Fgf8 dosage. Either eliminating or increasing Fgf8 expression increased apoptosis, whereas reducing Fgf8 expression had the opposite effect. To explain these counterintuitive observations, we suggest that an FGF8-dependent cell-survival pathway is negatively regulated by intracellular inhibitors produced in proportion to FGF8 concentration. Our data provide insight into the function of FGF8 in forebrain development and underscore the value of using multiple alleles and different experimental approaches to unravel the complexities of gene function in vertebrate development. PMID- 12574515 TI - Phenotype variation in two-locus mouse models of Hirschsprung disease: tissue specific interaction between Ret and Ednrb. AB - Clinical expression of Hirschsprung disease (HSCR) requires the interaction of multiple susceptibility genes. Molecular genetic analyses have revealed that interactions between mutations in the genes encoding the RET receptor tyrosine kinase and the endothelin receptor type B (EDNRB) are central to the genesis of HSCR. We have established two locus noncomplementation assays in mice, using allelic series at Ednrb in the context of Ret kinase-null heterozygotes, to understand the clinical presentation, incomplete penetrance, variation in length of aganglionic segment, and sex bias observed in human HSCR patients. Titration of Ednrb in the presence of half the genetic dose of Ret determines the presentation of an enteric phenotype in these strains, revealing or abrogating a sex bias in disease expression depending on the genotype at Ednrb. RET and EDNRB signaling pathways are also critical for the normal development of other tissues, including the kidneys and neural crest-derived melanocytes. Our data demonstrate that interaction between these genes is restricted to the enteric nervous system and does not affect renal, coat color, and retinal choroid development. PMID- 12574516 TI - Boosting immunity by antiviral drug therapy: a simple relationship among timing, efficacy, and success. AB - Drug therapies against persistent human infections such as hepatitis C virus, hepatitis B virus, and HIV fail to consistently eradicate the infection from the host. Hence, recent emphasis has shifted to the study of antiviral therapy aimed at boosting specific immune responses. It was argued that structured therapy interruptions were required to achieve this, because such regimes have shown promising results in early HIV infection. Using mathematical models, we show that, contrary to this notion, a single phase of drug therapy can result in the establishment of sustained immunity. We present a simple relationship between timing of therapy and efficacy of the drugs required for success. In the presence of strong viral suppression, we show that therapy should be stopped relatively early, and that a longer duration of treatment leads to failure. On the other hand, in the presence of weaker viral suppression, stopping treatment too early is detrimental, and therapy has to be continued beyond a time threshold. We discuss our modeling results primarily in the context of HCV therapy during chronic infection. Although the therapy regimes explored here also have implications for HIV, virus-mediated destruction of specific immune cells renders success unlikely during the chronic phase of the infection. PMID- 12574517 TI - Genome-wide protective response used by group A Streptococcus to evade destruction by human polymorphonuclear leukocytes. AB - Group A Streptococcus (GAS) evades polymorphonuclear leukocyte (PMN) phagocytosis and killing to cause human disease, including pharyngitis and necrotizing fasciitis (flesh-eating syndrome). We show that GAS genes differentially regulated during phagocytic interaction with human PMNs comprise a global pathogen-protective response to innate immunity. GAS prophage genes and genes involved in virulence, oxidative stress, cell wall biosynthesis, and gene regulation were up-regulated during PMN phagocytosis. Genes encoding novel secreted proteins were up-regulated, and the proteins were produced during human GAS infections. We discovered an essential role for the Ihk-Irr two-component regulatory system in evading PMN-mediated killing and promoting host-cell lysis, processes that would facilitate GAS pathogenesis. Importantly, the irr gene was highly expressed during human GAS pharyngitis. We conclude that a complex pathogen genetic program circumvents human innate immunity to promote disease. The gene regulatory program revealed by our studies identifies previously undescribed potential vaccine antigens and targets for therapeutic interventions designed to control GAS infections. PMID- 12574518 TI - Drosophila pigmentation evolution: divergent genotypes underlying convergent phenotypes. AB - Similar phenotypic changes have evolved independently in many animal taxa. It is unknown whether independent changes involve the same or different developmental and genetic mechanisms. Myriad pigment patterns in the genus Drosophila offer numerous opportunities to address this question. Previous studies identified regulatory and structural genes involved in the development and diversification of pigmentation in selected species. Here, we examine Drosophila americana and Drosophila novamexicana, interfertile species that have evolved dramatic pigmentation differences during the few million years since their divergence. Interspecific genetic analysis was used to investigate the contribution of five specific candidate genes and other genomic regions to phenotypic divergence by testing for associations between molecular markers and pigmentation. At least four distinct genomic regions contributed to pigmentation differences, one of which included the ebony gene. Ebony protein was expressed at higher levels in the more yellow D. novamexicana than the heavily melanized D. americana. Because Ebony promotes yellow pigment formation and suppresses melanization, the expression difference and genetic association suggest that evolution at the ebony locus contributed to pigmentation divergence between D. americana and D. novamexicana. Surprisingly, no genetic association with the yellow locus was detected in this study, and Yellow expression was identical in the two species. Evolution at the yellow locus underlies pigmentation divergence among other Drosophila species; thus, similar pigment patterns have evolved through regulatory changes in different genes in different lineages. These findings bear upon understanding classic models of melanism and mimicry. PMID- 12574520 TI - Direct chemical evidence for widespread dairying in prehistoric Britain. AB - Domesticated animals formed an important element of farming practices in prehistoric Britain, a fact revealed through the quantity and variety of animal bone typically found at archaeological sites. However, it is not known whether the ruminant animals were raised purely for their tissues (e.g., meat) or alternatively were exploited principally for their milk. Absorbed organic residues from pottery from 14 British prehistoric sites were investigated for evidence of the processing of dairy products. Our ability to detect dairy fats rests on the observation that the delta(13)C values of the C(18:0) fatty acids in ruminant dairy fats are approximately 2.3 per thousand lower than in ruminant adipose fats. This difference can be ascribed to (i) the inability of the mammary gland to biosynthesize C(18:0); (ii) the biohydrogenation of dietary unsaturated fatty acids in the rumen; and (iii) differences (i.e., 8.1 per thousand ) in the delta(13)C values of the plant dietary fatty acids and carbohydrates. The lipids from a total of 958 archaeological pottery vessels were extracted, and the compound-specific delta(13)C values of preserved fatty acids (C(16:0) and C(18:0)) were determined via gas chromatography-combustion-isotope ratio mass spectrometry. The results provide direct evidence for the exploitation of domesticated ruminant animals for dairy products at all Neolithic, Bronze Age, and Iron Age settlements in Britain. Most significantly, studies of pottery from a range of key early Neolithic sites confirmed that dairying was a widespread activity in this period and therefore probably well developed when farming was introduced into Britain in the fifth millennium B.C. PMID- 12574519 TI - Cloning, expression, and characterization of a membrane progestin receptor and evidence it is an intermediary in meiotic maturation of fish oocytes. AB - The structures of membrane receptors mediating rapid, nongenomic actions of steroids have not been identified. We describe the cloning of a cDNA from spotted seatrout ovaries encoding a protein that satisfies the following seven criteria for its designation as a steroid membrane receptor: plausible structure, tissue specificity, cellular distribution, steroid binding, signal transduction, hormonal regulation, and biological relevance. For plausible structure, computer modeling predicts that the protein has seven transmembrane domains, typical of G protein-coupled receptors. The mRNA (4.0 kb) is only detected in the brain and reproductive tissues on Northern blots. Antisera only detect the protein (40 kDa) in plasma membranes of reproductive tissues. The recombinant protein produced in an Escherichia coli expression system has a high affinity (K(d) = 30 nM), saturable, displaceable, single binding site specific for progestins. Progestins alter signal transduction pathways, activating mitogen-activated protein kinase and inhibiting adenylyl cyclase, in a transfected mammalian cell line. Inhibition of adenylyl cyclase is pertussis toxin sensitive, suggesting the receptor may be coupled to an inhibitory G protein. Progestins and gonadotropin up-regulate both mRNA and protein levels in seatrout ovaries. Changes in receptor abundance in response to hormones and at various stages of oocyte development, its probable coupling to an inhibitory G protein and inhibition of progestin induction of oocyte maturation upon microinjection of antisense oligonucleotides are consistent with the identity of the receptor as an intermediary in oocyte maturation. These characteristics suggest the fish protein is a membrane progestin receptor mediating a "nonclassical" action of progestins to induce oocyte maturation in fish. PMID- 12574521 TI - DNA duplex-quadruplex exchange as the basis for a nanomolecular machine. AB - There is currently great interest in the design of nanodevices that are capable of performing linear or rotary movements. Protein molecular machines are abundant in biology but it has recently been proposed that nucleic acids could also act as nanomolecular machines in model systems. Several types of movements have been described with DNA machines: rotation and "scissors-like" opening and closing. Here we show a nanomachine that is capable of an extension-contraction movement. The simple and robust device described here is composed of a single 21-base oligonucleotide and relies on a duplex-quadruplex equilibrium that may be fueled by the sequential addition of DNA single strands, generating a DNA duplex as a by product. The interconversion between two well defined topological states induces a 5-nm two-stroke, linear motor-type movement, which is detected by fluorescence resonance energy transfer spectroscopy. PMID- 12574522 TI - Using temporal context to improve biosurveillance. AB - Current efforts to detect covert bioterrorist attacks from increases in hospital visit rates are plagued by the unpredictable nature of these rates. Although many current systems evaluate hospital visit data 1 day at a time, we investigate evaluating multiple days at once to lessen the effects of this unpredictability and to improve both the timeliness and sensitivity of detection. To test this approach, we introduce simulated disease outbreaks of varying shapes, magnitudes, and durations into 10 years of historical daily visit data from a major tertiary care metropolitan teaching hospital. We then investigate the effectiveness of using multiday temporal filters for detecting these simulated outbreaks within the noisy environment of the historical visit data. Our results show that compared with the standard 1-day approach, the multiday detection approach significantly increases detection sensitivity and decreases latency while maintaining a high specificity. We conclude that current biosurveillance systems should incorporate a wider temporal context to improve their effectiveness. Furthermore, for increased robustness and performance, hybrid systems should be developed to capitalize on the complementary strengths of different types of temporal filters. PMID- 12574524 TI - Neurogenesis and apoptotic cell death. PMID- 12574526 TI - New light on white matter. PMID- 12574525 TI - Vascular protection. PMID- 12574527 TI - Infection, inflammation, and atherosclerosis. PMID- 12574528 TI - Converging pathogenic mechanisms in vascular and neurodegenerative dementia. PMID- 12574529 TI - Estrogen and stroke: a balanced analysis. PMID- 12574530 TI - Stroke genetics update. PMID- 12574531 TI - Stroke neuroimaging. PMID- 12574532 TI - Ever decreasing circles: advances in antiplatelet therapy and anticoagulation. PMID- 12574533 TI - Statins in threatened stroke. PMID- 12574534 TI - Angiotensin-converting enzyme inhibitors for stroke prevention: is there HOPE for PROGRESS After LIFE? PMID- 12574535 TI - Advances in interventional neuroradiology. PMID- 12574536 TI - Emerging therapies for acute ischemic stroke: new therapies on trial. PMID- 12574537 TI - Review of critical care and emergency approaches to stroke. PMID- 12574538 TI - Stroke recovery and rehabilitation. PMID- 12574539 TI - Prevention and health services delivery. PMID- 12574540 TI - Health policy in stroke: translating what we know into what we do. PMID- 12574541 TI - Controversy: the essence of medical debate. PMID- 12574542 TI - Contribution of Stroke to the Cochrane Stroke Group Trials Register. PMID- 12574543 TI - Clinical use of C-reactive protein for prognostic stratification in ischemic stroke: has the time come for including it in the patient risk profile? PMID- 12574544 TI - Postmortem MRI as a useful tool for investigation of cerebral microbleeds. PMID- 12574545 TI - Improving the assessment of outcomes in stroke: use of a structured interview to assign grades on the modified Rankin Scale. PMID- 12574546 TI - Risk of stroke associated with nonsteroidal anti-inflammatory drugs: a nested case-control study. AB - BACKGROUND AND PURPOSE: Nonsteroidal anti-inflammatory drugs (NSAIDs) have been associated with bleeding complications and may affect the risk of hemorrhagic stroke through inhibition of platelet cyclooxygenase-1. We performed a population based case-control study to estimate the risk of intracerebral hemorrhage, subarachnoid hemorrhage, and ischemic stroke in users of NSAIDs. METHODS: We used a population-based patient registry to identify all patients with a first-ever stroke discharge diagnosis in the period of 1994 to 1999. All diagnoses were validated according to predefined criteria. We selected 40 000 random controls from the background population. Information on drug use for cases and controls was retrieved from a prescription registry. Odds ratios were adjusted for age, sex, calendar year, and use of other medication. To evaluate the effect of various potential confounders not recorded in the register, we performed separate analyses on data from 2 large population-based surveys with more detailed information on risk factors. RESULTS: The cases were classified as intracerebral hemorrhage (n=659), subarachnoid hemorrhage (n=208), and ischemic stroke (n=2717). The adjusted odds ratio of stroke in current NSAID users compared with never users was 1.2 (95% CI, 0.9 to 1.6) for intracerebral hemorrhage, 1.2 (95% CI, 0.7 to 2.1) for subarachnoid hemorrhage and 1.2 (95% confidence interval, 1.0 to 1.4) for ischemic stroke. The survey data indicated that additional confounder control would not have led to an increase in relative risk estimates. CONCLUSIONS: Current exposure to NSAIDs is not a risk factor for intracerebral hemorrhage or subarachnoid hemorrhage. Furthermore, NSAIDs probably offer no protection against first-ever ischemic stroke. PMID- 12574547 TI - Nonaspirin nonsteroidal anti-inflammatory drugs and risk of hospitalization for intracerebral hemorrhage: a population-based case-control study. AB - BACKGROUND AND PURPOSE: Nonsteroidal anti-inflammatory drugs (NSAIDs) have effects on hemostasis and have been associated with an increased risk of bleeding. However, data relating the use of nonaspirin NSAIDs and risk of intracerebral hemorrhage (ICH) are sparse. METHODS: Using data from the County Hospital Patient Register and the Civil Registration System of North Jutland County, Denmark, we identified 912 cases of first-time ICH and 9059 sex- and age matched population-based controls in the period of 1991 to 1999. All prescriptions for nonaspirin NSAIDs before the date of admission for ICH were identified through a population-based prescription database. Conditional logistic regression was used to adjust for potential confounding factors, including previous discharge diagnoses of hypertension, chronic bronchitis and emphysema, alcoholism, liver cirrhosis, diabetes mellitus, and prescriptions for insulin or oral hypoglycemic agents, antihypertensive agents, lipid-lowering agents, low dose aspirin, high-dose aspirin, and oral anticoagulants. RESULTS: No overall association was found between prescription for nonaspirin NSAIDs in the preceding 30, 60, or 90 days and risk of ICH; ie, odds ratios ranged from 0.92 (95% CI, 0.70 to 1.21) to 1.13 (95% CI, 0.81 to 1.58). Furthermore, there was no increased risk of ICH associated with prescription for nonaspirin NSAIDs when the study population was stratified by age, sex, and a previous discharge diagnosis of hypertension. CONCLUSIONS: Patients prescribed nonaspirin NSAIDs were not at an overall increased risk of being hospitalized for ICH. This reassuring finding was seen in all examined subgroups, including the elderly and patients with a previous discharge diagnosis of hypertension. PMID- 12574548 TI - Incidence and risk factors of silent brain infarcts in the population-based Rotterdam Scan Study. AB - BACKGROUND AND PURPOSE: The prevalence of silent brain infarcts in healthy elderly people is high, and these lesions are associated with an increased risk of stroke. The incidence of silent brain infarcts is unknown. We investigated the incidence and cardiovascular risk factors for silent brain infarcts. METHODS: The Rotterdam Scan Study is a prospective, population-based cohort study of 1077 participants 60 to 90 years of age. All participants underwent cranial MRI in 1995 to 1996, and 668 participants had a second MRI in 1999 to 2000 (response rate, 70%) with a mean interval of 3.4 years. We assessed cardiovascular risk factors by interview and physical examination at baseline. Associations between risk factors and incident silent infarcts were analyzed by multiple logistic regression. RESULTS: Ninety-three participants (14%) had > or =1 new infarcts on the second MRI; of these, 81 had only silent and 12 had symptomatic infarcts. The incidence of silent brain infarcts strongly increased with age and was 5 times higher than that of symptomatic stroke. A prevalent silent brain infarct strongly predicted a new silent infarct on the second MRI (age- and sex-adjusted odds ratio, 2.9; 95% confidence interval, 1.7 to 5.0). Age, blood pressure, diabetes mellitus, cholesterol and homocysteine levels, intima-media thickness, carotid plaques, and smoking were associated with new silent brain infarcts in participants without prevalent infarcts. CONCLUSIONS: The incidence of silent brain infarcts on MRI in the general elderly population strongly increases with age. The cardiovascular risk factors for silent brain infarcts are similar to those for stroke. PMID- 12574549 TI - Genetic and environmental contributions to atherosclerosis phenotypes in men and women: heritability of carotid intima-media thickness in the Framingham Heart Study. AB - BACKGROUND AND PURPOSE: Carotid intima-media thickness (IMT) is a quantitative measure of subclinical atherosclerosis that is predictive of subsequent myocardial infarction and stroke. There is controversy regarding the proportion of variability in IMT explained by genetic factors. Thus, it is uncertain whether carotid IMT is a heritable trait that can be used in genetic studies. METHODS: From 1996 to 1998, we measured carotid IMT in 906 men (mean age, 56.7 years) and 980 women (mean age, 57.4 years) from 586 extended families (1630 sib pairs) in the Framingham Offspring cohort. B-mode carotid ultrasonography was used to define mean and maximum IMT of the common carotid artery (CCA) and internal carotid artery (ICA). Correlation coefficients were calculated in pairs of siblings. Variance component methods were used to estimate heritability with crude, age- and sex-adjusted, and multivariable-adjusted normalized deviates. RESULTS: Multivariable-adjusted correlation coefficients for mean CCA and ICA IMT were 0.16 and 0.16, respectively. Crude, age- and sex-adjusted, and multivariable adjusted heritabilities were 0.67, 0.44, and 0.38 for the mean CCA IMT (all P<0.001) and 0.43, 0.37, and 0.35 for the mean ICA IMT (all P<0.001). For CCA IMT, 27% of the overall variance was due to measured covariates; 38% was due to heritable factors. CONCLUSIONS: These data suggest that a substantial proportion of the variability in carotid IMT is explained by genetic factors. Further studies of genetic linkage and candidate gene association are warranted to identify specific genetic variants predisposing to subclinical atherosclerosis and stroke. PMID- 12574550 TI - Interleukin-6 promoter polymorphism modulates the effects of heavy alcohol consumption on early carotid artery atherosclerosis: the Carotid Atherosclerosis Progression Study (CAPS). AB - BACKGROUND AND PURPOSE: A J-shaped relationship has been demonstrated between alcohol and both clinical cardiovascular events and carotid atherosclerosis. A similar J-shaped relationship has been found between alcohol intake and inflammatory markers. If inflammation were on the intermediate causal pathway between alcohol intake and atherosclerosis, then genetic determinants of enhanced inflammation would be expected to modify this relationship. METHODS: In a large community population (n=1000; age, 50 to 65 years), we studied the effects of the interleukin-6 (IL-6)-174 polymorphism and gene-alcohol interactions on common carotid artery intima-media thickness (CCA-IMT) and carotid plaque. RESULTS: The CC genotype was associated with significantly higher IL-6 levels; the odds ratio (OR) for IL-6 in the top quartile was 2.07 (95% CI, 1.16 to 3.72; P=0.014). Interactions were seen between genotype and alcohol consumption for both IL-6 levels and CCA-IMT. In individuals who drank >30 g/d of alcohol, the CC genotype was associated with higher IL-6 levels, elevated CCA-IMT (P=0.001), and increased risk of carotid plaque (OR, 3.64; 95% CI, 1.15 to 11.54; P=0.028). The J-shaped relationship between alcohol intake and IMT was seen only for the CC genotype. CONCLUSIONS: These data suggest that the IL-6-174 promotor polymorphism may modulate the effects of alcohol on carotid atherosclerosis. These data support the hypothesis that inflammation forms part of the intermediate causal pathway between alcohol intake and atherosclerosis. PMID- 12574551 TI - Racial differences in thoracic aorta atherosclerosis among ischemic stroke patients. AB - BACKGROUND AND PURPOSE: Atherosclerosis of the thoracic aorta is an independent risk factor for stroke. There is little information on the impact of race in the prevalence of thoracic aorta atherosclerotic plaques among ischemic stroke patients. This study was an attempt to objectively assess the prevalence, thickness, and burden of thoracic aorta atherosclerotic plaques in a large population of ischemic stroke patients and to compare the differences between American blacks and whites. METHODS: This is a retrospective study of clinical data and transesophageal echocardiography (TEE) of 1553 ischemic stroke patients (664 blacks, 889 whites) over a period of 4.5 years. Atherosclerotic plaque prevalence, thickness, morphology, and burden (sum of maximum thickness in ascending aorta [AA], aortic arch [AO], and descending aorta [DA]) were assessed with TEE. Charts were reviewed for clinical information. RESULTS: Age and sex were similar among blacks and whites. Analyses of clinical data found that blacks had significantly higher hypertension (odds ratio [OR], 2.61; P<0.0001) and diabetes mellitus (OR, 1.99; P<0.0001) and significantly lower coronary artery disease (OR, 0.75; P=0.017) and carotid artery disease (OR, 0.62; P=0.0008) compared with whites. TEE showed that whites had significantly greater plaque prevalence (AA: OR, 1.37; P=0.04; AO: OR, 1.26; P=0.03; DA: OR, 1.39; P=0.002) and plaque burden (blacks, 4.28 mm; whites, 4.97 mm; P=0.007). Whites also had a trend of increased complex plaques and plaques >4 mm thick in all regions of the thoracic aorta. CONCLUSIONS: Among ischemic stroke patients, blacks had a lower prevalence of extra cranial atherosclerotic disease even though they had significantly higher hypertension and diabetes mellitus compared with whites. This difference cannot be explained by the existing risk factors in ischemic stroke patients. PMID- 12574552 TI - Atrial fibrillation and the prothrombotic state in the elderly: the Rotterdam Study. AB - BACKGROUND AND PURPOSE: Atrial fibrillation (AF) is a major cause of stroke among the elderly. Evidence for a prothrombotic state in AF is controversial, and there is a lack of studies among the elderly. We studied the relationships between AF and 3 prothrombotic plasma markers-von Willebrand factor (vWf; a marker of endothelial damage/dysfunction), soluble P-selectin (sP-sel; a marker of platelet activation), and fibrinogen-in a matched case-control study nested within a large community-based study of an elderly population. METHODS: We identified 162 elderly participants (mean+/-SD age, 78+/-8 years; 51% male) in the Rotterdam Study with documented AF and matched each case by age and sex to 2 population controls. vWf and sP-sel were measured by enzyme-linked immunosorbent assay; fibrinogen was measured with the Clauss method. We used conditional logistic regression analysis to assess the relationships between the markers and AF, adjusting for potential confounders. RESULTS: There were no significant relationships between either fibrinogen (P=0.8) or sP-sel (P=0.6) and AF. However, a positive linear relationship between vWf level and presence of AF remained significant after adjustment for potential confounders among women (odds ratio [OR], 1.17; 95% CI, 1.02 to 1.34) per 10-IU/dL increase in vWf but not among men (OR, 1.06; 95% CI, 0.96 to 1.17). CONCLUSIONS: We observed a positive relationship between AF and plasma vWf (or endothelial damage/dysfunction) in our elderly population, which was most apparent among women. Fibrinogen and sP-sel levels were unrelated to AF. The prothrombotic state of AF may be subject to sex differences, but longitudinal studies are needed to determine the relationship between these plasma markers and stroke risk. PMID- 12574553 TI - Mortality from cerebrovascular disease in a cohort of 23 000 patients with insulin-treated diabetes. AB - BACKGROUND AND PURPOSE: Disease of the cardiovascular system is the main cause of long-term complications and mortality in patients with type I (insulin-dependent) and type II (non-insulin-dependent) diabetes. Cerebrovascular mortality rates have been shown to be raised in patients with type II diabetes but have not previously been reported by age and sex in patients with type I diabetes. METHODS: A cohort of 23 751 patients with insulin-treated diabetes, diagnosed under the age of 30 years from throughout the United Kingdom, was identified during 1972 to 1993 and followed up for mortality until the end of December 2000. Age- and sex-specific mortality rates and standardized mortality ratios (SMRs) were calculated. RESULTS: There were 1437 deaths during the follow-up, 80 due to cerebrovascular disease. Overall, the cerebrovascular mortality rates in the cohort were higher than the corresponding rates in the general population, and the SMRs were 3.1 (95% CI, 2.2 to 4.3) for men and 4.4 (95% CI, 3.1 to 6.0) for women. When stratified by age, the SMRs were highest in the 20- to 39-year age group. After subdivision of cause of death into hemorrhagic and nonhemorrhagic origins, there remained a significant increase in mortality from stroke of nonhemorrhagic origin. CONCLUSIONS: Analyses of mortality from this cohort, essentially one of patients with type I diabetes, has shown for the first time that cerebrovascular mortality is raised at all ages in these patients. Type I diabetes is at least as great a risk factor for cerebrovascular mortality as type II diabetes. PMID- 12574554 TI - Family history and risk of subarachnoid hemorrhage: a case-control study in Nagoya, Japan. AB - BACKGROUND AND PURPOSE: We sought to examine the relation between a family history of subarachnoid hemorrhage (SAH) and the risk of SAH by using a case control study. METHODS: Case subjects consisted of a consecutive series of 195 patients with spontaneous SAH, aged 30 to 79 years, with aneurysms confirmed by angiography and/or CT scan. Hospital and community control subjects were identified and matched to each case by sex and age (+/-2 years). Multiple conditional logistic regression was used to calculate the odds ratio (OR) and 95% interval (CI) adjusted for potential confounders. RESULTS: Having a family member with SAH was significantly associated with an increased risk of SAH (OR, 4.0, 95% CI, 2.0 to 8.0), after adjusting for potential confounders. The risk for a positive family history of SAH was similar for men and women and was inversely related to the SAH patient's age. A maternal positive SAH history (OR, 5.4; 95% CI, 1.8 to 16.0) posed a much greater risk than a paternal positive history (OR, 3.2, 95% CI, 1.1 to 13.4). CONCLUSIONS: A positive family history of SAH was significantly and strongly associated with the risk of SAH. To prevent the onset of SAH at a younger age, much more attention should be given to individuals with any family member (first-degree relatives) suffering SAH episodes. PMID- 12574555 TI - Possible role for vascular cell proliferation in cerebral vasospasm after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: During vasospasm after subarachnoid hemorrhage (SAH), cerebral blood vessels show structural changes consistent with the actions of vascular mitogens. We measured platelet-derived vascular growth factors (PDGFs) in the cerebrospinal fluid (CSF) of patients after SAH and tested the effect of these factors on cerebral arteries in vivo and in vitro. METHODS: CSF was sampled from 14 patients after SAH, 6 patients not suffering SAH, and 8 normal controls. ELISA was performed for PDGF-AB, transforming growth factor-beta1, and vascular endothelial growth factor. A mouse model was used to compare cerebral vascular cell proliferation and PDGF staining in SAH compared with sham-operated controls. Normal human pial arteries were incubated for 7 days in vitro, 2 groups with human blood clot and 1 with and 1 without PDGF antibodies. RESULTS: PDGF-AB concentrations in CSF from SAH patients were significantly higher than those from non-SAH patients and normal controls, both during the first week after SAH and for all time points measured. Smooth muscle and fibroblast proliferation was observed after SAH in the mouse model, and this cellular replication was observed in conjunction with PDGF protein at the sites of thrombus. In human pial arteries, localized thrombus stimulated vessel wall proliferation, and proliferation was blocked by neutralizing antibodies directed against PDGFs. CONCLUSIONS: Vascular mitogens are increased in the CSF of patients after SAH. Proliferation of cells in the vascular wall is associated with perivascular thrombus. Cellular proliferation and subsequent vessel wall thickening may contribute to the syndrome of delayed cerebral vasospasm. PMID- 12574556 TI - Gene transfer of extracellular superoxide dismutase reduces cerebral vasospasm after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Superoxide may play an important role in cerebral vasospasm after subarachnoid hemorrhage (SAH). Our first goal was to determine the effect of gene transfer of extracellular superoxide dismutase (ECSOD) on vasospasm after experimental SAH. Our second goal was to determine whether tissue binding of ECSOD via the heparin-binding domain (HBD) is important for the effect of the enzyme. Thus, we examined effects of gene transfer of ECSOD with the HBD deleted (ECSODDeltaHBD). METHODS: Adenovirus expressing human ECSOD (AdECSOD), ECSODDeltaHBD (AdECSODDeltaHBD), or no transgene (AdBglII) was injected into the cisterna magna of anesthetized rabbits 30 minutes after induction of experimental SAH. Cerebral angiography, an assay for ECSOD activity in cerebrospinal fluid (CSF), and Western blotting for human ECSOD in the basilar artery were performed. RESULTS: Baseline diameter of the basilar artery averaged 0.77+/-0.01 mm (mean+/ SEM) and was similar in all treatment groups. Decreases in diameter of the basilar artery 2 days after SAH were smaller after AdECSOD (11+/-3%; n=10) than after AdBglII (25+/-4%; n=7; P<0.05). ECSOD activity was not detected in CSF before SAH and gene transfer. Of 8 rabbits treated with AdECSOD, in which ECSOD activity in CSF was measured after SAH, 5 animals had detectable ECSOD activity in CSF (46+/-13 U/mL). In these 5 rabbits, the diameter decreased by only 6+/-3%, and ECSOD protein was detected in the basilar artery. After AdECSODDeltaHBD (n=4), despite high levels of ECSOD activity in CSF (91+/-19 U/mL), vessel diameter decreased by 20+/-2%, and no ECSODDeltaHBD protein was detected in the basilar artery. CONCLUSIONS: Gene transfer of ECSOD reduces cerebral vasospasm after experimental SAH. Tissue binding of the enzyme is essential for cerebral vascular effects of ECSOD. PMID- 12574557 TI - Visual rating of age-related white matter changes on magnetic resonance imaging: scale comparison, interrater agreement, and correlations with quantitative measurements. AB - BACKGROUND AND PURPOSE: To provide further insight into the MRI assessment of age related white matter changes (ARWMCs) with visual rating scales, 3 raters with different levels of experience tested the interrater agreement and comparability of 3 widely used rating scales in a cross-sectional and follow-up setting. Furthermore, the correlation between visual ratings and quantitative volumetric measurement was assessed. METHODS: Three raters from different sites using 3 established rating scales (Manolio, Fazekas and Schmidt, Scheltens) evaluated 74 baseline and follow-up scans from 5 European centers. One investigator also rated baseline scans in a set of 255 participants of the Austrian Stroke Prevention Study (ASPS) and measured the volume of ARWMCs. RESULTS: The interrater agreement for the baseline investigation was fair to good for all scales (kappa values, 0.59 to 0.78). On the follow-up scans, all 3 raters depicted significant ARWMC progression; however, the direct interrater agreement for this task was poor (kappa, 0.19 to 0.39). Comparison of the interrater reliability between the 3 scales revealed a statistical significant difference between the scale of Manolio and that of Fazekas and Schmidt for the baseline investigation (z value, -2.9676; P=0.003), demonstrating better interrater agreement for the Fazekas and Schmidt scale. The rating results obtained with all 3 scales were highly correlated with each other (Spearman rank correlation, 0.712 to 0.806; P< or =0.01), and there was significant agreement between all 3 visual rating scales and the quantitative volumetric measurement of ARWMC (Kendall W, 0.37, 0.48, and 0.57; P<0.001). CONCLUSIONS: Our data demonstrate that the 3 rating scales studied reflect the actual volume of ARWMCs well. The 2 scales that provide more detailed information on ARWMCs seemed preferential compared with the 1 that yields more global information. The visual assessment of ARWMC progression remains problematic and may require modifications or extensions of existing rating scales. PMID- 12574558 TI - Effects of acute hyperhomocysteinemia on the neurovascular coupling mechanism in healthy young adults. AB - BACKGROUND AND PURPOSE: Hyperhomocysteinemia is a vascular risk factor that infers with the nitric oxide signaling pathway of endothelial vasoregulation. Most investigations in young healthy humans on the peripheral vasculature using a standardized methionine challenge demonstrated altered vascular reactivity. In contrast, the cerebral autoregulation mechanism was shown to be unaffected by the same methionine load. To obtain additional insight into the compensatory range of the cerebral vasculature during a methionine challenge, we tested the neurovascular coupling mechanism that adjusts cerebral blood flow in accordance with cortical activity. METHODS: Fifteen healthy young adults (age, 24.7+/-2.3 years; 7 men) were tested with a functional transcranial Doppler test before and 3, 8, and 24 hours after administration of placebo, 20 mg folic acid, 20 mg folic acid and 0.1 g/kg body weight L-methionine, or L-methionine alone. Evoked blood flow response was evaluated according to a control system approach. Plasma concentrations of homocysteine, resting blood flow velocities, and control system parameters of flow velocity change were compared for each time point using a multiple analysis of variance test. RESULTS: Homocysteine levels increased significantly compared with baseline (before, 7.6+/-1.9 micromol/L; 3 hours, 22.2+/-6.0 micromol/L [P<0.0001]; 8 hours, 27.9+/-8.6 micromol/L [P<0.0001]; 24 hours, 12.6+/-7.8 micro mol/L [P=NS]). Resting flow velocities and control system parameters remained statistically nonsignificant. CONCLUSIONS: Compared with the peripheral vasculature, the regulatory mechanisms controlling adequate cerebral blood flow appear to have a wider compensatory range. This is concluded from statistically nonsignificant results comparing the vascular reactivity in young adults undergoing a standardized methionine challenge. Our data confirm indirectly the reports of high concentrations of homocysteine needed to affect the cerebral vasculature in animal experiments. PMID- 12574559 TI - Infection and risk of ischemic stroke: differences among stroke subtypes. AB - BACKGROUND AND PURPOSE: Although prior studies have demonstrated that 25% to 35% of stroke patients have had a recent infection, the role of infection as a risk factor remains unclear. Our aim was to characterize the effect of infectious/inflammatory syndromes on stroke risk. METHODS: Case-control and crossover analyses of 233 cases and 363 controls aged 21 to 89 years were performed. Cases were patients hospitalized with a first ischemic stroke at a Los Angeles, California, medical center. Controls were outpatients in the hypertension, diabetes, and general medical clinics. All subjects were administered a neurological examination, an infection/inflammation (I/I) examination, and an interview to elicit recent I/I history at baseline (within several days of stroke onset) and again approximately 2 months later. Three physicians classified subjects by the presence or absence of I/I within 1 month of the index dates, based on findings of the I/I examination, the interview report, and laboratory results. RESULTS: Infections, either total or specific, were not found more frequently in cases than controls. However, patients with a recent respiratory tract infection suffered more often from large-vessel atherothromboembolic or cardioembolic stroke than did patients without infection (48% vs 24%, P=0.07). The age- and sex-adjusted relative risk estimate for these subtypes was 1.75 (95% CI, 0.86 to 3.55). The risk was notably high for those without stroke risk factors: 4.15 (95% CI, 1.22 to 14.1) for normotensives, 2.71 (95% CI, 1.04 to 7.06) for nondiabetics, and 1.74 (95% CI, 0.74 to 4.07) for nonsmokers. Patients with a recent respiratory infection also had a more severe neurological deficit on admission than those without infection (P=0.05). CONCLUSIONS: Our results suggest that respiratory tract infection may act as a trigger and increase the risk of large-vessel and/or cardioembolic ischemic stroke, especially in those without vascular risk factors. PMID- 12574560 TI - Baseline magnetic resonance imaging parameters and stroke outcome in patients treated by intravenous tissue plasminogen activator. AB - BACKGROUND AND PURPOSE: We designed a prospective sequential pretreatment and posttreatment MRI study to assess the relation between neuroimaging parameters and clinical outcome in patients treated with intravenous recombinant tissue-type plasminogen activator (rtPA). METHODS: Patients with symptoms of acute hemispheric ischemic stroke were recruited. The National Institutes of Health Stroke Scale (NIHSS) score was assessed at baseline and at days 1, 7, and 60, and the modified Rankin scale (mRS) at day 60, by which outcome was classified in terms of independence (mRS score 0, 1, or 2) or severe disability or death (mRS score 3 through 6), was assigned. Multimodal stroke MRI was performed at presentation and repeated at day 1. MRI procedures included magnetic resonance angiography, T2* gradient-echo sequence, echoplanar imaging, and isotropic diffusion- (DWI) and perfusion-weighted (PWI) imaging. Patients were treated with intravenous rtPA after MRI completion. RESULTS: Twenty-nine patients (16 men and 13 women; mean+/-SD age, 65+/-14 years) underwent MRI; the mean time from symptom onset to treatment was 255+/-62 minutes. Twenty-six patients had a vessel occlusion, and 15 patients experienced a partial (Thrombolysis in Myocardial Infarction [TIMI]-2) or total (TIMI-3) recanalization at day 1, whereas 11 patients had a persistent occlusion. Mean NIHSS scores at day 60 were 5.7+/-5.4 if recanalization had occurred and 14+/-2 in cases of persistent occlusion. According to the mRS, 13 patients were independent (mRS 0 through 2), whereas severe disability or death (mRS 3 through 6) was observed in 15 patients. A better outcome was observed when recanalization was achieved (r=-0.68, P=0.0002). PWI volume and time to peak (TTP) within the DWI lesion assessed before therapy were correlated with day-60 NIHSS score (PWI volume: r=0.51, P=0.006, TTP: r=0.35, P=0.07). The day-0 DWI abnormality volume was well correlated with day-60 NIHSS score (r=0.58, P=0.001). Multiple regression linear analysis showed that 2 factors mainly influenced clinical outcome: (1) recanalization, with a high correlation with NIHSS score at day 60 (P=0.0001) and (2) day-0 DWI lesion volume, which is closely associated with day-60 NIHSS score (P=0.03). CONCLUSIONS: Baseline DWI volume and recanalization are the main factors influencing clinical outcome after rtPA for ischemic stroke. PMID- 12574561 TI - Are some patients likely to benefit from recombinant tissue-type plasminogen activator for acute ischemic stroke even beyond 3 hours from symptom onset? AB - BACKGROUND AND PURPOSE: Recombinant tissue plasminogen activator (rtPA) has been demonstrated to improve outcomes in acute ischemic stroke when delivered within 3 hours of symptom onset. However, the Alteplase Thrombolysis for Acute Noninterventional Therapy in Ischemic Stroke (ATLANTIS B) trial, in which patients were treated mostly between 3 and 5 hours after symptom onset, found no overall benefit from rtPA. We hypothesized that a subgroup of patients at low risk for thrombolysis-related intracranial hemorrhage, identifiable on the basis of pretreatment clinical variables, may benefit even when treated after 3 hours, despite the overall results of the trial. METHODS: Using an independently derived multivariate model that predicts the risk of thrombolysis-related intracranial hemorrhage in patients receiving tPA for acute myocardial infarction (based on 6 easily obtainable clinical characteristics), we stratified patients in the ATLANTIS B trial into low-, intermediate-, and high-risk tertiles. We examined outcomes in the prespecified low-risk subgroup using a global test of significance across 4 outcome scales. RESULTS: Despite having a similar average baseline stroke severity and median time to treatment (270 minutes), patients in the prespecified low-risk group (n=194) were significantly less likely to have a symptomatic intracranial hemorrhage than other patients in the trial (2.2% versus 9.2%, P=0.03). Although there was no treatment effect for rtPA in the overall trial, a consistent trend favoring rtPA therapy (a 5% to 12% absolute treatment benefit) was found across 4 different stroke scales in the prespecified low-risk group (P=0.10). The treatment-benefit-by-risk interaction was significant (P=0.03). CONCLUSIONS: Use of a multivariate index based on clinical variables is a promising approach to assist in the selection of patients with a favorable risk benefit profile for thrombolytic therapy beyond 3 hours. PMID- 12574562 TI - Inhibition of factor Xa reduces ischemic brain damage after thromboembolic stroke in rats. AB - BACKGROUND AND PURPOSE: Factor Xa (FXa) is a key coagulation protease and target for novel antithrombotic agents for prevention and treatment of diverse thromboembolic disorders. In the present study we describe the effect of a novel, potent, and selective FXa inhibitor, DPC602, on brain damage and neurobehavioral consequence in a rat thromboembolic model of stroke. METHODS: Thromboembolic stroke was induced in rats by placement of an autologous clot into the middle cerebral artery. RESULTS: Laser-Doppler monitoring of cerebral blood flow demonstrated that DPC602 (8 mg/kg, single IV/IP bolus pretreatment) markedly improved cerebral blood flow after thromboembolic stroke by 25% to 160% (n=6; P<0.001) at 1 to 6 hours. DPC602 demonstrated concentration- and time-dependent reductions in infarct size, with maximal effect (89% reduction; n=14; P<0.001) at the highest dose over controls. Neurological function was also significantly improved in DPC602-treated rats at days 1, 3, and 7 (n=13; P<0.01). DPC602 treatment did not cause cerebral hemorrhage, assessed by free hemoglobin in the ischemic brain tissues. CONCLUSIONS: These data suggest that anticoagulation with a selective FXa inhibitor might ameliorate the extent of ischemic brain damage and neurological deficits after a thromboembolic event. Enhanced clot dissolution and early reperfusion may account for the cerebrovascular-protective effect of the drug. PMID- 12574563 TI - Safety of dexamphetamine in acute ischemic stroke: a randomized, double-blind, controlled dose-escalation trial. AB - BACKGROUND AND PURPOSE: Amphetamine is reported to enhance recovery after experimental stroke, but results from the first few trials in humans are inconclusive. Limited information is available on treatment safety. This study intended to investigate the safety and tolerability of dexamphetamine in patients with acute cerebral ischemia. METHODS: Forty-five patients with cerebral ischemia were enrolled within 72 hours after onset of symptoms. Patients were randomized to 1 of 3 dose levels (2.5, 5, or 10 mg orally twice daily) or placebo for 5 consecutive days. Adverse events, blood pressure, heart rate, body temperature, consciousness level, and functional outcome measures were followed up daily during treatment. Follow-ups were made at day 7 and 1 and 3 months after stroke. RESULTS: Mean systolic and diastolic blood pressures and heart rate increased 14 mm Hg, 8 mm Hg, and 9 bpm, respectively, with dexamphetamine treatment compared with placebo (P< or =0.01). There was no difference between dexamphetamine and placebo regarding adverse events, body temperature, or consciousness level. During treatment, there was a benefit of dexamphetamine in neurological and functional outcome (P<0.05), but differences were not maintained at follow-up. CONCLUSIONS: Overall, dexamphetamine was safe and well tolerated by patients with acute cerebral ischemia, but blood pressure and heart rate increased during treatment in comparison to placebo. These observations may be important in the future planning of amphetamine trials because changes in these variables have been observed to interfere with clinical outcome. The suggestions of improvement in outcome must be confirmed in further studies. PMID- 12574564 TI - Tolerability of NXY-059 at higher target concentrations in patients with acute stroke. AB - BACKGROUND AND PURPOSE: NXY-059 is a nitrone-based free radical-trapping agent in development for acute stroke. In patients with acute stroke, NXY-059 is well tolerated at concentrations known to be associated with neuroprotection in animal models of transient cerebral ischemia; however, higher target concentrations appear necessary on the basis of animal models of permanent ischemia. METHODS: This was a randomized, double-blind, placebo-controlled, parallel-group, dose escalation, multicenter study that evaluated safety, tolerability, and plasma concentrations of 2 NXY-059 dosing regimens within 24 hours of acute stroke. NXY 059 was administered as either 915 mg over 1 hour followed by 420 mg/h for 71 hours or 1820 mg for 1 hour followed by 844 mg/h for 71 hours; plasma concentrations were monitored. Neurological and functional outcomes were recorded for up to 30 days. RESULTS: One hundred thirty-five patients were recruited, of whom 134 received study treatment and completed assessments (844 mg/h, n=39; 420 mg/h, n=48; placebo, n=47). Mean age was 69 years (range, 34 to 92 years), and baseline National Institutes of Health Stroke Scale score was 8.5 (SD, 6.6). Serious adverse events occurred in 3, 17, and 13 patients, respectively, with deaths in 0, 4, and 3 patients and treatment discontinuations because of adverse events in 0, 1, and 3 patients. Good outcome, defined by modified Rankin Scale score of 0 or 1, was seen in 53%, 29% and 40%, respectively. No safety concern was identified in analysis of body temperature, blood pressure, or other laboratory parameters. The unbound plasma concentration at steady state was 260+/ 79 micromol/L, exceeding the target of 200 micromol/L in the high-dose group. CONCLUSIONS: NXY-059 was well tolerated in patients with an acute stroke at and above concentrations shown to be neuroprotective in an animal model when initiated 4 hours after onset of permanent focal ischemia. PMID- 12574565 TI - Physical and social functioning after stroke: comparison of the Stroke Impact Scale and Short Form-36. AB - BACKGROUND AND PURPOSE: This study evaluated assessments of physical functioning and social functioning using the Stroke Impact Scale (SIS) and Short Form-36 (SF 36) to characterize health-related quality of life for patients after stroke. METHODS: The SIS and SF-36 were administered to 278 stroke subjects approximately 90 days after stroke. The SIS-16 and SF-36 Physical Functioning (PF) domain characterize physical function, whereas the SIS Participation and SF-36 Social Functioning (SF) domains characterize social function. Descriptive statistics and an analysis of variance were used to characterize physical and social functioning after stroke across levels of the modified Rankin Scale (MRS). Rasch analysis was used to compare the hierarchies and ranges of item difficulties in the SIS-16 and the SF-36 PF domains, as well as in the SIS Participation and the SF-36 SF domains. RESULTS: Item hierarchies for the SIS-16 and SF-36 PF domain demonstrate that the SIS-16 contains less difficult items that could differentiate physical function among patients with more severe limitations. Compared with the SF-36 SF domain, the item hierarchy for the SIS Participation domain contained more difficult items that could differentiate social function among patients who were more active. In contrast to SIS-16, the SF-36 PF has major floor effects. In contrast to SIS Participation, the SF-36 SF domain has major ceiling effects. Both SIS-16 and SF-36 PF were able to discriminate well among the MRS levels of 0 to 1, 2, 3, and 4. The SIS Participation domain was also able to discriminate across the MRS levels of 0 to 1, 2, and 3 to 4. On the other hand, the SF-36 SF was similar among MRS levels 0, 1, and 2 and among MRS levels 2, 3, and 4. CONCLUSIONS: Both the physical and participation subscales of the SIS cover a wider range of item difficulty than their counterparts from the SF-36. Compared with the SF-36 PF and SF domains, the SIS-16 and SIS Participation are better able to capture physical functioning and social well-being in patients with strokes. PMID- 12574566 TI - Risk factors for falling in home-dwelling older women with stroke: the Women's Health and Aging Study. AB - BACKGROUND AND PURPOSE: Much of our knowledge of risk factors for falls comes from studies of the general population. The aim of this study was to estimate the risk of falling associated with commonly accepted and stroke-specific factors in a home-dwelling stroke population. METHODS: This study included an analysis of prospective fall reports in 124 women with confirmed stroke over 1 year. Variables relating to physical and mental health, history of falls, stroke symptoms, self-reported difficulties in activities of daily living, and physical performance tests were collected during home assessments. RESULTS: Risk factors for falling commonly reported in the general population, including performance tests of balance, incontinence, previous falls, and sedative/hypnotic medications, did not predict falls in multivariate analyses. Frequent balance problems while dressing were the strongest risk factor for falls (odds ratio, 7.0). Residual balance, dizziness, or spinning stroke symptoms were also a strong risk factor for falling (odds ratio, 5.2). Residual motor symptoms were not associated with an increased risk of falling. CONCLUSIONS: Interventions to reduce the frequency of balance problems during complex tasks may play a significant role in reducing falls in stroke. Clinicians should be aware of the increased risk of falling in women with residual balance, dizziness, or spinning stroke symptoms and recognize that risk assessments developed for use in the general population may not be appropriate for stroke patients. PMID- 12574567 TI - Measuring longitudinally the metabolic demands of stroke patients: resting energy expenditure is not elevated. AB - BACKGROUND AND PURPOSE: Little is known of the acute, subacute, and longer-term energy demands of stroke, information essential to appropriate clinical and nutritional management. The goals of this study were to (1) determine the resting energy expenditure (REE) of stroke patients from stroke onset to 3 months, (2) examine relations between stroke size, type, location, severity, and REE, and (3) evaluate whether estimation of REE from the Harris-Benedict equation (HB) requires the addition of a "stress factor" to capture the possible additional REE imposed by stroke. METHODS: The REE of new stroke patients was measured prospectively at hospital admission and on days 7, 11, 14, 21, and 90 by indirect calorimetry. Stroke patients' REEs (Kcal/d) over time and REEs as a percentage of HB were compared with control subjects' single measurements. RESULTS: Mean REE and %HB of stroke patients ranged from 1521+/-290 to 1663+/-268 Kcal/d and from 107+/-14.9 to 114+/-12.9 %HB, respectively. Mean measurements of control subjects were 1665+/-265 Kcal/d and 112.9+/-11.4 %HB (NS). REE was not associated with stroke characteristics (NS). Changes in REE measured longitudinally were not clinically meaningful (4 to 62 Kcal/d) though statistically significant (P=0.004). CONCLUSIONS: The REEs of stroke patients and controls were both approximately 10% higher than those predicted by HB. No hypermetabolic response pattern of energy expenditure was evident after stroke. REE did not vary with stroke characteristics, although confirmation with larger subgroups is required. PMID- 12574568 TI - Lower endotoxin immunity predicts increased cognitive dysfunction in elderly patients after cardiac surgery. AB - BACKGROUND AND PURPOSE: Although coronary artery bypass graft surgery (CABG) improves the quality of life and functional capacity for numerous patients, many also exhibit impairment in cognitive function immediately after surgery. Although the etiology of this cognitive decline is multifactorial, the inflammatory response to the primary insult may modulate the extent of dysfunction. Patients with low preoperative levels of anti-endotoxin core antibody (EndoCAb) are more likely to experience adverse outcomes, suggesting that decreased immunity to endotoxin causes a heightened release of inflammatory mediators. We therefore sought to determine the association of decreased EndoCAb and the incidence of postoperative cognitive decline. METHODS: EndoCAb levels were measured before surgery in 460 patients undergoing elective CABG. Cognitive function was measured preoperatively and 6 weeks postoperatively. Multivariable analysis accounted for the effects of age, Parsonnet score, sex, body mass index, baseline cognition, years of education, history of hypertension, bypass time, cross-clamp time, and number of grafts. RESULTS: At 6-week follow-up, 122 patients (36%) showed cognitive decline. Lower preoperative EndoCAb levels were associated with a greater incidence and severity of postoperative cognitive decline. The elderly with decreased endotoxin immunity are particularly susceptible to this decline (relative risk=1.97 for age >64). CONCLUSIONS: Reduced preoperative endotoxin immunity is a predictor of increased postoperative cognitive dysfunction in patients undergoing CABG, particularly in those >60 years old. Interventions that increase IgM EndoCAb levels might improve cognitive function after cardiac surgery. PMID- 12574569 TI - Reanalysis of the final results of the European Carotid Surgery Trial. AB - BACKGROUND AND PURPOSE: The European Carotid Surgery Trial (ECST) and North American Symptomatic Carotid Endarterectomy Trial (NASCET) have shown that endarterectomy reduces the risk of stroke in certain patients with recently symptomatic carotid stenosis. However, they differed in the degree of stenosis above which surgery was reported to be effective. This disparity has led to inconsistent clinical recommendations but may have been due to differences between the trials in the methods of measurement of carotid stenosis and definitions of outcome events. METHODS: To allow direct comparison of analyses from ECST and NASCET, we remeasured the prerandomization ECST carotid angiograms and redefined the outcome events the same way as in NASCET. RESULTS: We randomized 3018 patients and followed them up for a mean of 73 months. Surgery reduced the 5-year risk of any stroke or surgical death by 5.7% (95% CI, 0 to 11.6) in patients with 50% to 69% stenosis (n=646, P=0.05) and by 21.2% (95% CI, 12.9 to 29.4) in patients with 70% to 99% stenosis without "near occlusion" (n=429, P<0.0001). These benefits were maintained at the 10-year follow-up. However, surgery was of no benefit in patients (n=125) with near occlusion. The effect of surgery in this group was highly significantly different from that in patients with 70% to 99% stenosis without near occlusion (P=0.002). Surgery was harmful in patients with <30% stenosis (n=1321, P=0.007) and of no benefit in patients with 30% to 49% stenosis (n=478, P=0.6). CONCLUSIONS: Results of the ECST and NASCET were consistent when analyzed in the same way. In ECST, surgery was highly beneficial for 70% to 99% stenosis and moderately beneficial for 50% to 69% stenosis. However, contrary to clinical recommendations and current practice, surgery was of little benefit in patients with carotid near occlusion. PMID- 12574570 TI - Patient selection for carotid endarterectomy: how far is risk modeling applicable to the individual? AB - BACKGROUND AND PURPOSE: Risk-factor modeling has been proposed to identify patients with carotid stenosis who will most benefit from surgery. Validation by independent institutions performing carotid endarterectomy is necessary to determine the applicability of such models to the individual patient. METHODS: A series of patients with a recently symptomatic high-grade carotid stenosis were selected for surgery according to current guidelines and were consecutively operated on in a single institution. In addition, a prognostic model was applied to the patients to analyze the concordance of both selection methods. RESULTS: The study included 134 patients operated on between 1999 and 2001. The risk model predicted that 49% of the patients should have been excluded from surgery because the operation was found to be possibly harmful in 1 patient (1%) and not significantly beneficial in 65 patients (48%). This resulted from the predominant negative weight of the surgical risk factors in the model. However, this predominance was negated in our series by the fact that only 1 major complication (0.75%) occurred during follow-up. CONCLUSIONS: Exclusion of single patients on the basis of risk modeling may be problematic when the rate of perioperative complications is very low. PMID- 12574571 TI - Costs and caregiver consequences of early supported discharge for stroke patients. AB - BACKGROUND AND PURPOSE: Early supported discharge (ESD) for stroke has been shown to yield outcomes similar to or better than those of conventional care, but there is less information on the impact on costs and on the caregiver. The purpose of this study is to estimate the costs associated with an ESD program compared with those of usual care. METHODS: We conducted a randomized controlled trial of stroke patients who required rehabilitation services and who had a caregiver at home. RESULTS: Acute-care costs incurred before randomization when patients were medically ready for discharge averaged $3251 per person. The costs for the balance of the acute-care stay, from randomization to discharge, were $1383 for the home group and $2220 for the usual care group. The average cost of providing the 4-week home intervention service was $943 per person. The total cost generated by persons assigned to the home group averaged $7784 per person, significantly lower than the $11 065 per person for those assigned to usual care. A large proportion of the cost differential between the 2 groups arose from readmissions, for which the usual care group generated costs more than quadruple those of the home intervention group. CONCLUSIONS: Providing care at home was no more (or less) expensive for those with greater functional limitation than for those with less. Caregivers in the ESD group scored consistently lower on the Burden Index than caregivers with usual care, even caregivers of persons with major functional limitations. For persons recovering from stroke and their families, ESD provides a cost-effective alternative to usual care. PMID- 12574572 TI - Vampire bat salivary plasminogen activator (desmoteplase): a unique fibrinolytic enzyme that does not promote neurodegeneration. AB - BACKGROUND AND PURPOSE: Tissue-type plasminogen activator (tPA) promotes excitotoxic and ischemic injury within the brain. These findings have implications for the use of tPA in the treatment of acute ischemic stroke. The plasminogen activator from vampire bat (Desmodus rotundus) saliva (D rotundus salivary plasminogen activator [DSPA]; desmoteplase) is an effective plasminogen activator but, in contrast to tPA, is nearly inactive in the absence of a fibrin cofactor. The purpose of this study was to compare the ability of DSPA and tPA to promote kainate- and N-methyl-D-aspartate (NMDA)-induced neurodegeneration in tPA /- mice and wild-type mice, respectively. METHODS: tPA-/- mice were infused intracerebrally with either tPA or DSPA. The degree of neuronal survival after hippocampal injection of kainate was assessed histochemically. Wild-type mice were used to assess the extent of neuronal damage after intrastriatal injection of NMDA in the presence of tPA or DSPA. Immunohistochemistry and fibrin zymography were used to evaluate DSPA and tPA antigen or activity. RESULTS: Infusion of tPA into tPA-/- mice restored sensitivity to kainate-mediated neurotoxicity and activation of microglia. DSPA was incapable of conferring sensitivity to kainate treatment, even when infused at 10-fold higher molar concentration than tPA. The presence of tPA also increased the lesion volume induced by NMDA injection into the striatum of wild-type mice, whereas DSPA had no effect. CONCLUSIONS: DSPA does not promote kainate- or NMDA-mediated neurotoxicity in vivo. These results provide significant impetus to evaluate DSPA in patients with ischemic stroke. PMID- 12574573 TI - Essential role of Gap junctions in NO- and prostanoid-independent relaxations evoked by acetylcholine in rabbit intracerebral arteries. AB - BACKGROUND AND PURPOSE: Direct intercellular communication via gap junctions may play a central role in endothelium-dependent relaxations that are mediated by a conducted hyperpolarization and do not involve the synthesis of NO and prostanoids. In the present study, inhibitory peptides homologous to the Gap27 domain of the second extracellular loop of connexin37/connexin43 and connexin40, designated as 37,43Gap27 and 40Gap27, respectively, were used to evaluate the role of this mechanism in intracerebral arteries. METHODS: Isolated rings of rabbit middle cerebral artery were constricted by histamine (10 micromol/L) in the presence of N(G)-nitro-L-arginine methyl ester (300 micromol/L) and indomethacin (10 micromol/L). Concentration-relaxation curves for acetylcholine were constructed in the presence and absence of 37,43Gap27 and 40Gap27. Specific antibodies were used to delineate the distribution of connexin37, connexin40, connexin43, and connexin45 within the arterial wall. RESULTS: Individually, 37,43Gap27 and 40Gap27 minimally affected endothelium-dependent relaxations to acetylcholine at concentrations of 300 micro mol/L, whereas their combination (at 300 micromol/L each) inhibited the maximal response by approximately 70% and increased the EC50 value for relaxation by approximately 15-fold. In endothelium denuded rings, this peptide combination did not attenuate responses to sodium nitroprusside, an exogenous source of NO. Gap junction plaques, whose incidence was highest in endothelium, were constructed from connexin40 and connexin43 in the media and connexin37, connexin40, and connexin43 in the endothelium. CONCLUSIONS: The findings confirm that direct communication via gap junctions contributes to agonist-induced relaxations of intracerebral arteries. More than one connexin subtype appears to participate in such responses. PMID- 12574574 TI - Withdrawal of statin treatment abrogates stroke protection in mice. AB - BACKGROUND AND PURPOSE: Statins (3-hydroxy-3-methylglutaryl-coenzyme A [HMG-CoA] reductase inhibitors) reduce stroke damage independent of lipid lowering by upregulation of endothelial nitric oxide synthase (eNOS). Acute withdrawal of statin treatment may suppress endothelial NO production and impair vascular function. METHODS: To test this hypothesis, we treated 129/SV mice with atorvastatin (10 mg/kg) for 14 days and then withdrew treatment. RESULTS: Treatment with atorvastatin conferred stroke protection by 40% after filamentous occlusion of the middle cerebral artery followed by reperfusion. Withdrawal of statin treatment, however, resulted in the loss of stroke protection after 2 and 4 days. In mouse aortas and brain vasculature, statins upregulated eNOS message 2.3- and 1.7-fold, respectively, as measured by reverse transcription-polymerase chain reaction. Withdrawal of statins resulted in 5- and 2.7-fold downregulation of eNOS in aorta and brain, respectively, after 2 days. Statin treatment decreased RhoA GTPase membrane expression to 48%, while withdrawal of statins resulted in 4-fold increase of RhoA in the membrane. Moreover, platelet factor 4 and beta-thromboglobulin in plasma were significantly downregulated by statin treatment, but withdrawal of statins resulted in a 2.9- and 3.1-fold upregulation after 2 days, respectively. Thrombus formation induced by ligature of the inferior vena cava was significantly reduced by statin treatment. When statin treatment was withdrawn, however, protection was lost between 2 and 4 days. CONCLUSIONS: Acute termination of statin treatment results in a rapid loss of protection in mouse models of cerebral ischemia and thrombus formation independent of lipid lowering. In patients with acute or impending stroke, withdrawal of statins may impair outcome. PMID- 12574575 TI - Intravenous administration of bone morphogenetic protein-7 after ischemia improves motor function in stroke rats. AB - BACKGROUND AND PURPOSE: We and others have previously reported that bone morphogenetic protein-7 (BMP-7), given before middle cerebral artery occlusion (MCAO), reduces ischemic injury in brain. Recent studies have indicated that receptors for BMP are upregulated after brain ischemia. It is possible that this upregulation may facilitate endogenous neurorepair in the ischemic brain. The purpose of this study was to determine the neuroregenerative effects of BMP-7 given parenterally after ischemia/reperfusion injury. METHODS: Adult Sprague Dawley rats were anesthetized with chloral hydrate. The middle cerebral artery was transiently occluded by a filament inserted through the right internal carotid artery. The filament was removed after 60-minute ischemia to allow reperfusion. Some animals were killed 24 hours after MCAO to examine BMP-7 mRNA expression. Other animals received a single dose of intravenous BMP-7 or vehicle at 24 hours after MCAO and were used for subsequent behavioral studies and BMP-7 immunostaining. RESULTS: BMP-7 mRNA was upregulated 24 hours after MCAO in untreated animals. BMP-7 immunoreactivity was dose-dependently increased on the ischemic side of the hippocampus/dentate on day 6 after MCAO in animals receiving intravenous injection of BMP-7. Animals receiving BMP-7 also showed a decrease in body asymmetry from day 7 to day 14 and an increase in locomotor activity on day 14 after MCAO. CONCLUSIONS: Our data indicate that BMP-7, given parenterally after stroke, can pass through the blood-brain barrier on the ischemic side and induce behavioral recovery in stroke animals at longer testing times. PMID- 12574576 TI - Interobserver agreement in the classification of stroke in the Women's Health Study. AB - BACKGROUND AND PURPOSE: Accurate classification of stroke events is essential in large cohort studies of risk factor assessment and in treatment trials such as the Women's Health Study (WHS). METHODS: Based on medical record review, we assessed interrater reliability in stroke classification and disability status in the WHS using the kappa statistic. RESULTS: During 7.0 years of follow-up, 271 incident strokes occurred, of which 133 were reclassified. There was excellent interrater agreement in the diagnosis of major stroke types, hemorrhagic subtypes, and degree of disability, as well as substantial agreement in the definition of the vascular territory involved. Only moderate agreement was reached in the classification of ischemic subtypes. CONCLUSIONS: Major stroke types, degree of disability, and vascular territory involved can be reliably classified on the basis of a review of medical records in the WHS, whereas classification of ischemic stroke subtypes needs further refinement of diagnostic criteria. PMID- 12574577 TI - Modified National Institutes of Health Stroke Scale can be estimated from medical records. AB - BACKGROUND AND PURPOSE: The 15-item National Institutes of Health Stroke Scale (NIHSS) is a quantitative measure of stroke-related neurological deficit with established reliability and validity for use in clinical research. An abridged 11 item modified NIHSS (mNIHSS) has been described that simplifies or eliminates redundant and less reliable items. We aimed to determine whether the mNIHSS could be accurately abstracted from medical records to facilitate retrospective research. METHODS: We selected 39 patient records for which NIHSS scores were formally measured. Handwritten notes from medical records were abstracted, and NIHSS item scores were estimated by 5 raters blinded to actual scores. Estimated scores were compared among raters and with actual measured scores. RESULTS: Interrater reliability for total NIHSS on admission and discharge was excellent, with intraclass correlation coefficients (ICCs) of 0.85 and 0.79, respectively. However, ICCs for 2 items (facial palsy and dysarthria) were poor (<0.40). Interrater reliability for total mNIHSS was slightly greater, with ICCs of 0.87 and 0.89 on admission and discharge, respectively. None of the 11 mNIHSS items had poor reliability, 4 were moderate (ICC, 0.40 to 0.75), and 7 were excellent (ICC >0.75). Sixty-two percent of estimated total NIHSS scores were within 2 points of actual scores and 91% were within 5 points, whereas 70% of estimated total mNIHSS scores were within 2 points and 95% were within 5 points. CONCLUSIONS: The mNIHSS can be estimated from medical records with a high degree of reliability and validity. In retrospective assessment of stroke severity, the mNIHSS performs better than the standard NIHSS and may be easier to use because it has fewer and simpler items. PMID- 12574578 TI - Hyperbaric oxygen therapy in acute ischemic stroke: results of the Hyperbaric Oxygen in Acute Ischemic Stroke Trial Pilot Study. AB - BACKGROUND AND PURPOSE: Hyperbaric oxygen therapy (HBO) has promise as a treatment for acute stroke. This study was conducted to evaluate the efficacy, safety, and feasibility of using HBO in acute ischemic stroke. METHODS: We conducted a randomized, prospective, double-blind, sham-controlled pilot study of 33 patients presenting with acute ischemic stroke who did not receive thrombolytics over a 24-month period. Patients were randomized to treatment for 60 minutes in a monoplace hyperbaric chamber pressurized with 100% O2 to 2.5-atm absolute (ATA) in the HBO group or 1.14 ATA in the sham group. Primary outcomes measured included percentage of patients with improvement at 24 hours (National Institutes of Health Stroke Scale [NIHSS]) and 90 days (NIHSS, Barthel Index, modified Rankin Scale, Glasgow Outcome Scale). Secondary measurements included complications of treatment and mortality at 90 days. RESULTS: Baseline demographics were similar in both groups. There were no differences between the groups at 24 hours (P=0.44). At 3 months, however, a larger percentage of the sham patients had a good outcome defined by their stroke scores compared with the HBO group (NIHSS, 80% versus 31.3%; P=0.04; Barthel Index, 81.8% versus 50%; P=0.12; modified Rankin Scale, 81.8% versus 31.3%; P=0.02; Glasgow Outcome Scale, 90.9% versus 37.5%; P=0.01) with loss of statistical significance in a intent-to treat analysis. CONCLUSIONS: Although our HBO protocol appears feasible and safe, it does not appear to be beneficial and may be harmful in patients with acute ischemic stroke. PMID- 12574579 TI - Imaging-based decision making in thrombolytic therapy for ischemic stroke: present status. AB - BACKGROUND: Thrombolysis is the treatment of choice for acute stroke within 3 hours after symptom onset. Treatment beyond the 3-hour time window has not been shown to be effective in any single trial; however, meta-analyses suggest a somewhat lesser but still significant effect within 3 to 6 hours after stroke. It seems reasonable to apply improved selection criteria that allow differentiation between patients with and without a relevant indication for thrombolytic therapy. SUMMARY OF REVIEW: The present literature on imaging in stroke has been thoroughly reviewed, covering Doppler ultrasound (DU), arteriography, CT, and MRI and including modern techniques such as perfusion CT, diffusion- and perfusion weighted MRI (DWI, PWI), CT angiography and MR angiography (CTA, MRA), and CTA source image analysis (CTA-SI). The authors present their view of a comprehensive diagnostic approach to acute stroke, which challenges the concept of a rigid therapeutic time window. CONCLUSIONS: Information about the presence or absence of a vessel occlusion, whether by means of DU, CTA, or MRA, is essential before recombinant tissue plasminogen activator is given in the 3- to 6-hour time window. Clear demarcation of the irreversibly damaged infarct core and the ischemic but still viable and thus salvageable tissue at risk of infarction as seen on DWI/PWI/MRA or alternatively CT/CTA/CTA-SI should be obtained before thrombolysis is initiated within 3 to 6 hours. Once these advanced techniques are used, the therapeutic time window can be extended with acceptable safety. However, comprehensive informed consent is mandatory, especially when thrombolytic therapy is considered beyond established time windows. PMID- 12574580 TI - Acetylcholinesterase inhibitors for vascular dementia and Alzheimer's disease combined with cerebrovascular disease. PMID- 12574581 TI - In-hospital care pathways for stroke: a Cochrane systematic review. PMID- 12574582 TI - Major ongoing stroke trials. PMID- 12574583 TI - The ironic politics of obesity. PMID- 12574584 TI - Space program. Shuttle disaster puts NASA plans in tailspin. PMID- 12574585 TI - Space program. Scientists mourn many losses. PMID- 12574586 TI - Human cloning. New players, same debate in Congress. PMID- 12574587 TI - Public health. Bush pledges $15 billion to combat AIDS in 14 countries. PMID- 12574589 TI - ITER. United States rejoins international fusion-research project. PMID- 12574588 TI - India. Panel finds plagiarism by university leader. PMID- 12574590 TI - Forestry. Old-growth forest spared for now. PMID- 12574591 TI - United Kingdom. Walport to take reins at Wellcome Trust. PMID- 12574592 TI - National labs. IBM scientist to lead Brookhaven. PMID- 12574593 TI - Geology. Efforts to tame second African 'killer lake' begin. PMID- 12574594 TI - 2004 budget request. Protecting the homeland sets tone for 2004 budget. PMID- 12574595 TI - 2004 budget request. NIH shuffles budget to prop up research. PMID- 12574596 TI - 2004 budget request. NSF is not yet seeing double. PMID- 12574597 TI - World Health Organization. Health body taps a consummate insider and disease fighter. PMID- 12574598 TI - Singapore. Novartis kicks off institute for neglected diseases. PMID- 12574599 TI - Biosafety labs. The architect behind the new fortresses of science. PMID- 12574600 TI - Biosafety labs. Space suits with that 'je ne sais quoi'. PMID- 12574601 TI - Biosafety labs. The war on bioterror moves west. PMID- 12574602 TI - Patents and the research exemption. PMID- 12574603 TI - Biopolymers and the environment. PMID- 12574606 TI - Perceptions of science. Face values: how portraits win friends and influence people. PMID- 12574604 TI - Comment on "Buffered Tree Population Changes in a Quaternary Refugium: Evolutionary Implications". PMID- 12574607 TI - Chemistry. Chemical reactions involving quantum tunneling. PMID- 12574608 TI - Space physics. Coupling across many scales. PMID- 12574609 TI - Biomedicine. Love, honor, and protect (your liver). PMID- 12574610 TI - Computational mathematics. Full steam ahead-probably. PMID- 12574611 TI - Cell biology. NO says yes to mitochondria. PMID- 12574612 TI - Materials science. Oxygen in crystals--seeing is believing. PMID- 12574613 TI - Physics. Coherent power booster. PMID- 12574615 TI - Cellular warriors at the battle of the bulge. PMID- 12574616 TI - Obesity drug pipeline not so fat. PMID- 12574617 TI - Having it all. PMID- 12574618 TI - Obesity and the environment: where do we go from here? AB - The obesity epidemic shows no signs of abating. There is an urgent need to push back against the environmental forces that are producing gradual weight gain in the population. Using data from national surveys, we estimate that affecting energy balance by 100 kilocalories per day (by a combination of reductions in energy intake and increases in physical activity) could prevent weight gain in most of the population. This can be achieved by small changes in behavior, such as 15 minutes per day of walking or eating a few less bites at each meal. Having a specific behavioral target for the prevention of weight gain may be key to arresting the obesity epidemic. PMID- 12574619 TI - A war on obesity, not the obese. AB - In their efforts to lose weight, obese individuals may be fighting a powerful set of evolutionary forces honed in an environment drastically different from that of today. PMID- 12574620 TI - A clinical view of the obesity problem. AB - The increased rate of obesity in the past decade is caused by a larger rate of energy input than energy expenditure. But this simple explanation belies the complexity of the possible solutions: Education about obesity needs to improve for both the medical community and the public. Successful treatment will require reforms of medical practice as well as more research to facilitate the development of new drugs. PMID- 12574621 TI - Potential symbionts in bathyal foraminifera. PMID- 12574622 TI - Au20: a tetrahedral cluster. AB - Photoelectron spectroscopy revealed that a 20-atom gold cluster has an extremely large energy gap, which is even greater than that of C60, and an electron affinity comparable with that of C60. This observation suggests that the Au20 cluster should be highly stable and chemically inert. Using relativistic density functional calculations, we found that Au20 possesses a tetrahedral structure, which is a fragment of the face-centered cubic lattice of bulk gold with a small structural relaxation. Au20 is thus a unique molecule with atomic packing similar to that of bulk gold but with very different properties. PMID- 12574623 TI - Carbon tunneling from a single quantum state. AB - We observed ring expansion of 1-methylcyclobutylfluorocarbene at 8 kelvin, a reaction that involves carbon tunneling. The measured rate constants were 4.0 x 10(-6) per second in nitrogen and 4 x 10(-5) per second in argon. Calculations indicated that at this temperature the reaction proceeds from a single quantum state of the reactant so that the computed rate constant has achieved a temperature-independent limit. According to calculations, the tunneling contribution to the rate is 152 orders of magnitude greater than the contribution from passage over the barrier. We discuss environmental effects of the solid state inert-gas matrix on the reaction rate. PMID- 12574624 TI - Atomic-resolution imaging of oxygen in perovskite ceramics. AB - Using an imaging mode based on the adjustment of a negative value of the spherical-aberration coefficient of the objective lens of a transmission electron microscope, we successfully imaged all types of atomic columns in the dielectric SrTiO3 and the superconductor YBa2Cu3O7. In particular, we were able to view the oxygen atoms which, due to their low scattering power, were not previously accessible, and this allowed us to detect local nonstoichiometries or the degree of oxygen-vacancy ordering. This technique offers interesting opportunities for research into oxides, minerals, and ceramics. In particular, this holds for the huge group of perovskite-derived electroceramic materials in which the local oxygen content sensitively controls the electronic properties. PMID- 12574625 TI - Formation of electron holes and particle energization during magnetic reconnection. AB - Three-dimensional particle simulations of magnetic reconnection reveal the development of turbulence driven by intense electron beams that form near the magnetic x-line and separatrices. The turbulence collapses into localized three dimensional nonlinear structures in which the electron density is depleted. The predicted structure of these electron holes compares favorably with satellite observations at Earth's magnetopause. The birth and death of these electron holes and their associated intense electric fields lead to strong electron scattering and energization, whose understanding is critical to explaining why magnetic explosions in space release energy so quickly and produce such a large number of energetic electrons. PMID- 12574626 TI - Eocene El Nino: evidence for robust tropical dynamics in the "hothouse". AB - Much uncertainty surrounds the interactions between the El Nino-Southern Oscillation (ENSO) and long-term global change. Past periods of extreme global warmth, exemplified by the Eocene (55 to 35 million years ago), provide a good testing ground for theories for this interaction. Here, we compare Eocene coupled climate model simulations with annually resolved variability records preserved in lake sediments. The simulations show Pacific deep-ocean and high-latitude surface warming of approximately 10 degrees C but little change in the tropical thermocline structure, atmosphere-ocean dynamics, and ENSO, in agreement with proxies. This result contrasts with theories linking past and future "hothouse" climates with a shift toward a permanent El Nino-like state. PMID- 12574627 TI - Lipid-like material as the source of the uncharacterized organic carbon in the ocean? AB - The composition and formation mechanisms of the uncharacterized fraction of oceanic particulate organic carbon (POC) are not well understood. We isolated biologically important compound classes and the acid-insoluble fraction, a proxy of the uncharacterized fraction, from sinking POC in the deep Northeast Pacific and measured carbon isotope ratios to constrain the source(s) of the uncharacterized fraction. Stable carbon and radiocarbon isotope signatures of the acid-insoluble fraction were similar to those of the lipid fraction, implying that the acid-insoluble fraction might be composed of selectively accumulated lipid-like macromolecules. PMID- 12574628 TI - Design and chemical synthesis of a homogeneous polymer-modified erythropoiesis protein. AB - We report the design and total chemical synthesis of "synthetic erythropoiesis protein" (SEP), a 51-kilodalton protein-polymer construct consisting of a 166 amino-acid polypeptide chain and two covalently attached, branched, and monodisperse polymer moieties that are negatively charged. The ability to control the chemistry allowed us to synthesize a macromolecule of precisely defined covalent structure. SEP was homogeneous as shown by high-resolution analytical techniques, with a mass of 50,825 +/-10 daltons by electrospray mass spectrometry, and with a pI of 5.0. In cell and animal assays for erythropoiesis, SEP displayed potent biological activity and had significantly prolonged duration of action in vivo. These chemical methods are a powerful tool in the rational design of protein constructs with potential therapeutic applications. PMID- 12574629 TI - Myc-induced T cell leukemia in transgenic zebrafish. AB - The zebrafish is an attractive model organism for studying cancer development because of its genetic accessibility. Here we describe the induction of clonally derived T cell acute lymphoblastic leukemia in transgenic zebrafish expressing mouse c-myc under control of the zebrafish Rag2 promoter. Visualization of leukemic cells expressing a chimeric transgene encoding Myc fused to green fluorescent protein (GFP) revealed that leukemias arose in the thymus, spread locally into gill arches and retro-orbital soft tissue, and then disseminated into skeletal muscle and abdominal organs. Leukemic cells homed back to the thymus in irradiated fish transplanted with GFP-labeled leukemic lymphoblasts. This transgenic model provides a platform for drug screens and for genetic screens aimed at identifying mutations that suppress or enhance c-myc- induced carcinogenesis. PMID- 12574630 TI - Angiogenesis-independent endothelial protection of liver: role of VEGFR-1. AB - The vascular endothelium was once thought to function primarily in nutrient and oxygen delivery, but recent evidence suggests that it may play a broader role in tissue homeostasis. To explore the role of sinusoidal endothelial cells (LSECs) in the adult liver, we studied the effects of vascular endothelial growth factor (VEGF) receptor activation on mouse hepatocyte growth. Delivery of VEGF-A increased liver mass in mice but did not stimulate growth of hepatocytes in vitro, unless LSECs were also present in the culture. Hepatocyte growth factor (HGF) was identified as one of the LSEC-derived paracrine mediators promoting hepatocyte growth. Selective activation of VEGF receptor-1 (VEGFR-1) stimulated hepatocyte but not endothelial proliferation in vivo and reduced liver damage in mice exposed to a hepatotoxin. Thus, VEGFR-1 agonists may have therapeutic potential for preservation of organ function in certain liver disorders. PMID- 12574631 TI - Inhibition of hepatitis B virus replication by drug-induced depletion of nucleocapsids. AB - Chronic hepatitis B virus (HBV) infection is a major cause of liver disease. Only interferon-alpha and the nucleosidic inhibitors of the viral polymerase, 3TC and adefovir, are approved for therapy. However, these therapies are limited by the side effects of interferon and the substantial resistance of the virus to nucleosidic inhibitors. Potent new antiviral compounds suitable for monotherapy or combination therapy are highly desired. We describe non-nucleosidic inhibitors of HBV nucleocapsid maturation that possess in vitro and in vivo antiviral activity. These inhibitors have potential for future therapeutic regimens to combat chronic HBV infection. PMID- 12574632 TI - Mitochondrial biogenesis in mammals: the role of endogenous nitric oxide. AB - Nitric oxide was found to trigger mitochondrial biogenesis in cells as diverse as brown adipocytes and 3T3-L1, U937, and HeLa cells. This effect of nitric oxide was dependent on guanosine 3',5'-monophosphate (cGMP) and was mediated by the induction of peroxisome proliferator-activated receptor gamma coactivator 1alpha, a master regulator of mitochondrial biogenesis. Moreover, the mitochondrial biogenesis induced by exposure to cold was markedly reduced in brown adipose tissue of endothelial nitric oxide synthase null-mutant (eNOS-/-) mice, which had a reduced metabolic rate and accelerated weight gain as compared to wild-type mice. Thus, a nitric oxide-cGMP-dependent pathway controls mitochondrial biogenesis and body energy balance. PMID- 12574633 TI - Yield effects of genetically modified crops in developing countries. AB - Onfarm field trials carried out with Bacillus thuringiensis (Bt) cotton in different states of India show that the technology substantially reduces pest damage and increases yields. The yield gains are much higher than what has been reported for other countries where genetically modified crops were used mostly to replace and enhance chemical pest control. In many developing countries, small scale farmers especially suffer big pest-related yield losses because of technical and economic constraints. Pest-resistant genetically modified crops can contribute to increased yields and agricultural growth in those situations, as the case of Bt cotton in India demonstrates. PMID- 12574634 TI - GUN4, a regulator of chlorophyll synthesis and intracellular signaling. AB - Nuclear genes control plastid differentiation in response to developmental signals, environmental signals, and retrograde signals from plastids themselves. In return, plastids emit signals that are essential for proper expression of many nuclear photosynthetic genes. Accumulation of magnesium-protoporphyrin IX (Mg Proto), an intermediate in chlorophyll biosynthesis, is a plastid signal that represses nuclear transcription through a signaling pathway that, in Arabidopsis, requires the GUN4 gene. GUN4 binds the product and substrate of Mg- chelatase, an enzyme that produces Mg-Proto, and activates Mg-chelatase. Thus, GUN4 participates in plastid-to-nucleus signaling by regulating Mg-Proto synthesis or trafficking. PMID- 12574635 TI - Passage of heme-iron across the envelope of Staphylococcus aureus. AB - The cell wall envelope of Gram-positive pathogens functions as a scaffold for the attachment of virulence factors and as a sieve that prevents diffusion of molecules. Here the isd genes (iron-regulated surface determinant) of Staphylococcus aureus were found to encode factors responsible for hemoglobin binding and passage of heme-iron to the cytoplasm, where it acts as an essential nutrient. Heme-iron passage required two sortases that tether Isd proteins to unique locations within the cell wall. Thus, Isd appears to act as an import apparatus that uses cell wall-anchored proteins to relay heme-iron across the bacterial envelope. PMID- 12574636 TI - Li15Al3Si6 (Li14.6Al3.4Si6), a compound displaying a heterographite-like anionic framework. AB - The title compound, lithium aluminium silicide (15/3/6), crystallizes in the hexagonal centrosymmetric space group P6(3)/m. The three-dimensional structure of this ternary compound may be depicted as two interpenetrating lattices, namely a graphite-like Li(3)Al(3)Si(6) layer and a distorted diamond-like lithium lattice. As is commonly found for LiAl alloys, the Li and Al atoms are found to share some crystallographic sites. The diamond-like lattice is built up of Li cations, and the graphite-like anionic layer is composed of Si, Al and Li atoms in which Si and Al are covalently bonded [Si-Al = 2.4672 (4) A]. PMID- 12574637 TI - Barium-deficient celsian, Ba1-xAl2-2xSi2+2xO8 (x = 0.20 or 0.06). AB - Barium-deficient forms of celsian (barium aluminium silicate) with the formula Ba(1-x)Al(2-2x)Si(2+2x)O(8) (x = 0.20 and 0.06) have been identified. In contrast with the celsian-orthoclase solid solutions which have been reported previously, these forms, refined in the space group C2/m, with Ba and one O atom in the 4i sites with m site symmetry, and a further O atom in a 4g site with twofold axial symmetry, suggest a slight solid solution with silica. The serendipitous preparation of the compounds represents a possible hazard associated with solid state synthesis. PMID- 12574638 TI - Perovskite-related LaTiO3.41. AB - Crystals of pentalanthanum pentatitanium heptadecaoxide (La(5)Ti(5)O(17) with 0.3% oxygen excess, or LaTiO(3.41)) have been synthesized by floating-zone melting, and the structure has been solved using single-crystal X-ray diffraction intensities. The monoclinic (P2(1)/c) structure consists of perovskite-like slabs of vertex-sharing TiO(6) octahedra, which are separated by additional oxygen layers. The slabs are five octahedra wide. Due to the adjustment of the TiO(6) octahedra to meet the coordination requirements of the La(3+) cations, a superstructure develops along the a axis. PMID- 12574639 TI - NaCa4Nb5O17: a layered perovskite AnBnO3n+2 compound. AB - Sodium tetracalcium pentaniobium heptadecaoxide, NaCa(4)Nb(5)O(17), corresponds to the n = 5 term of the homologous A(n)B(n)O(3n+2) family of structures composed of ABX(3) perovskite layers. The structure consists of perovskite-type blocks of n = 5 layers of NbO(6) octahedra, separated by an interblock region. Successive blocks along the b axis are displaced by 1/2c with respect to each other. The deformation of the NbO(6) octahedra is a minimum at the middle of each block, and increases along the direction of the b axis to a maximum at each end of the block. Ca and Na share the same intrablock sites, coordinated by 12 O atoms, whereas only Ca atoms are found in the interblock cavities, at sites with different coordination geometries. PMID- 12574640 TI - Three hydrates of the bisphosphonate risedronate, consisting of one molecular and two ionic structures. AB - Three different hydrates of risedronate were obtained by varying the pH of a solution containing the compound. At the pH values used, the N atom of the pyridine group is protonated and the compounds are zwitterionic. Crystals obtained directly from the synthesis resulted in risedronate monohydrate, or [1 hydroxy-1-phosphono-2-(pyridinium-3-yl)ethyl]phosphonate monohydrate, C(7)H(11)NO(7)P(2).H(2)O, (I), in which just one phosphonate group is negatively charged. Recrystallizations at pH values of 2 and 4 yielded risedronate dihydrate, or sodium [1-hydroxy-2-(pyridinium-3-yl)ethane-1,2 diyl]bis(phosphonate) dihydrate, Na(+).C(7)H(10)NO(7)P(2)(-).2H(2)O, (II). Finally, recrystallizations at pH values of 7 and 8 produced risedronate 2.5 hydrate, or sodium [1-hydroxy-2-(pyridinium-3-yl)ethane-1,2-diyl]bis(phosphonate) 2.5-hydrate, Na(+).C(7)H(10)NO(7)P(2)(-).2.5H(2)O, (III). At these four pH values, both phosphonate groups in (II) and (III) are negatively charged and coordinated to an Na(+) ion. Crystals of (II), i.e. those grown at pH values of 2 and 4, have isomorphous polymeric ion aggregate structures with geminal phosphonate and alcohol groups coordinated to the same Na(+) ion. On the other hand, crystals of (III), i.e. those grown at pH values of 7 and 8, have isomorphism polymeric ion aggregate structures with geminal phosphonate and alcohol groups coordinated to different Na(+) ions. PMID- 12574641 TI - The trans influence of the pyridine ligand on ruthenium(II)-porphyrin-carbene complexes. AB - In the two ruthenium(II)-porphyrin-carbene complexes (dibenzoylcarbenyl kappaC)(pyridine-kappaN)(5,10,15,20-tetra-p-tolylporphyrinato kappa(4)N)ruthenium(II), [Ru(C(15)H(10)O(2))(C(5)H(5)N)(C(48)H(36)N(4))], (I), and (pyridine-kappaN)(5,10,15,20-tetra-p-tolylporphyrinato-kappa(4)N)[bis(3 trifluoromethylphenyl)carbenyl-kappaC]ruthenium(II), [Ru(C(15)H(8)F(6))(C(5)H(5)N)(C(48)H(36)N(4))], (II), the pyridine ligand coordinates to the octahedral Ru atom trans with respect to the carbene ligand. The C(carbene)-Ru-N(pyridine) bonds in (I) coincide with a crystallographic twofold axis. The Ru-C bond lengths of 1.877 (8) and 1.868 (3) A in (I) and (II), respectively, are slightly longer than those of other ruthenium(II)-porphyrin carbene complexes, owing to the trans influence of the pyridine ligands. PMID- 12574642 TI - (E)-2-(2-adamantylidene)-3-(1-ferrocenylethylidene)succinic anhydride. AB - The title compound, [Fe(C(5)H(5))(C(21)H(21)O(3))], was obtained from successive Stobbe condensations between ketones and dimethyl succinate. The succinic anhydride five-membered ring is distorted significantly from planarity, with the butadiene moiety being twisted by 49.3 (2) degrees from planarity and the C atoms at the succinic anhydride end of the alkene bonds showing significant pyramidalization. The cyclopentadiene rings of the ferrocenyl moiety adopt an almost eclipsed conformation. PMID- 12574643 TI - A zinc-lithium complex of 4,7-bis(2-aminoethyl)-1,4,7-triazacyclononane-1 acetate. AB - The asymmetric unit of ([4,7-bis(2-aminoethyl)-1,4,7-triazacyclononan-1 yl]acetato)zinc(II) triaqua(mu-[4,7-bis(2-aminoethyl)-1,4,7-triazacyclononan-1 yl]acetato)lithium(I)zinc(II) chloride diperchlorate, [Zn(C(12)H(26)N(5)O(2))][LiZn(C(12)H(26)N(5)O(2))(H(2)O)(3)]Cl(ClO(4))(2), obtained from the reaction between the lithium salt of 4,7-bis(2-aminoethyl) 1,4,7-triazacyclononane-1-acetate and Zn(ClO(4))(2), contains two Zn(II) complexes in which each Zn(II) ion is six-coordinated by five N-atom donors and one O-atom donor from the ligand. One carboxylate O-atom donor is not involved in coordination to a Zn(II) atom, but coordinates to an Li+ ion, the tetrahedral geometry of Li+ being completed by three water molecules. The two complexes are linked via a hydrogen bond between a primary amine N-H group and the carboxylate O atom not involved in coordination to a metal. PMID- 12574644 TI - Dichloro(isopropylamido)bis(isopropylamine)(isopropylimido)tantalum(V), a monomeric TaV compound with imido, amido and amino moieties. AB - The title compound, [Ta(C(3)H(7)N)(C(3)H(8)N)Cl(2)(C(3)H(9)N)(2)], is the first monomeric example of a metal complex that features imido, amido and amino moieties in the same molecule. The Ta atom has distorted octahedral coordination, with the imido moiety trans to chlorine and the pseudo-axial ligands bent away from the imido moiety. Principal dimensions include Ta=N = 1.763 (8) A, Ta-N(H) = 1.964 (7) A, and Ta-N(H(2)) = 2.247 (7) and 2.262 (7) A. PMID- 12574645 TI - Bis[mu-P,P'-methylenebis(diisopropyl phosphonate) kappa3O,O':O]bis[chlorolithium(I)]: a centrosymmetric dimer of lithium chloride with terminal chloride anions. AB - The molecule of the title dimeric compound, [Li(2)Cl(2)(C(13)H(30)O(6)P(2))(2)] or [LiCl([((i)PrO)(2)P(O)](2)CH(2))](2), lies about an inversion center and features tetrahedrally coordinated Li atoms. The neutral ligands each chelate to one metal center and bridge to the other center through P=O units. Unusually for lithium chloride complexes, the Cl- ions are in terminal rather than bridging positions. Principal dimensions include Li-O(four-membered ring) = 1.959 (3) and 2.056 (3) A, Li-O(phosphonate ring) = 1.929 (3) A, and Li-Cl = 2.293 (3) A. PMID- 12574646 TI - Tetra-mu-alpha-alanine-bis[tetraaquagadolinium(III)] hexaperchlorate. AB - The title complex, [Gd(2)(C(3)H(7)NO(2))(4)(H(2)O)(8)](ClO(4))(6), contains centrosymmetric dimeric [Gd(2)(Ala)(4)(H(2)O)(8)](6+) cations (Ala is alpha alanine) and perchlorate anions. The four alanine molecules act as bridging ligands linking two Gd(3+) ions through their carboxylate O atoms. Each Gd(3+) ion is also coordinated by four water molecules, which complete an eightfold coordination in a square-antiprism fashion. The perchlorate anions and the methyl groups of the alanine ligands are disordered. PMID- 12574647 TI - Ferrocene compounds. XXXIV. 1'-(tert-butoxycarbonylamino)ferrocene-1-carboxylic acid. AB - Heteroannularly substituted ferrocene derivatives can act as model systems for various hydrogen-bonded assemblies of biomolecules formed, for instance, by means of O-H...O and N-H...O hydrogen bonding. The crystal structure analysis of 1' (tert-butoxycarbonylamino)ferrocene-1-carboxylic acid, [Fe(C(10)H(14)NO(2))(C(6)H(5)O(2))] or (C(5)H(4)COOH)Fe(C(5)H(4)NHCOOC(CH(3))(3), reveals two independent molecules within the asymmetric unit, and these are joined into discrete dimers by two types of intermolecular hydrogen bonds, viz. O H...O and N-H...O. The -COOH and -NHCOOR groups are archetypes for dimer formation via two eight-membered rings. The O-H...O hydrogen bonds [2.656 (3) and 2.663 (3) A] form a cyclic carboxylic acid dimer motif. Another eight-membered ring is formed by N-H...O hydrogen bonds [2.827 (3) and 2.854 (3) A] between the N-H group and an O atom of another carbamoyl moiety. The dimers are assembled in a herring-bone fashion in the bc plane. PMID- 12574648 TI - The CoIII-C bond in (1-thia-4,7-diazacyclodecyl-kappa3N4,N7,C10)(1,4,7 triazacyclononane-kappa3N1,N4,N7)cobalt(III) dithionate hydrate. AB - In the title compound, [Co(C(6)H(15)N(3))(C(7)H(15)N(2)S)]S(2)O(6).H(2)O, the Co C bond distance is 1.9930 (13) A, which is shorter than for related compounds with the linear 1,6-diamino-3-thiahexan-4-ide anion in place of the macrocyclic 1 thia-4,7-diazacyclodecan-8-ide anion. The coordinated carbanion produces an elongation of 0.102 (7) A of the Co-N bond to the 1,4,7-triazacyclononane N atom in the trans position. This relatively small trans influence is presumably a result of the triamine ligand forming strong bonds to the Co(III) atom. PMID- 12574649 TI - Bis(2,6-bis[3-(2,4,6-trimethylphenyl)pyrazol-1-yl-kappaN2]pyridine kappaN)cobalt(II) dinitrate at 290 and 150 K. AB - The title compound, [Co(C(29)H(29)N(5))(2)](NO(3))(2), contains a six-coordinate high-spin Co(II) ion with approximate local D(2d) symmetry. The bond lengths and angles at cobalt undergo only small changes between the two temperatures, which confirms that the Co(II) ion does not undergo a spin-state transition over this temperature range. PMID- 12574650 TI - Bis(azido-kappaN)[(6R,7S,8S,14S)-(-)-sparteine-kappa2N,N']copper(II). AB - In the title compound, [Cu(N(3))(2)(C(15)H(26)N(2))], the chiral alkaloid (-)-l sparteine (Sp) acts as a bidentate ligand, with two azide ligands occupying the remaining coordination sites, forming a distorted CuN(4) tetrahedron. The dihedral angle between the N(Sp)-Cu-N(Sp) and N(azide)-Cu-N(azide) planes is 55.3 (2) degrees. Principal dimensions include Cu-N(Sp) = 2.011 (6) and 2.025 (5) A, and Cu-N(azide) = 1.939 (6) and 1.934 (7) A. The mid-point of the N(Sp)...N(Sp) line does not lie exactly in the N(azide)-Cu-N(azide) plane, but is tilted towards one of the N(Sp) atoms by 0.026 A. PMID- 12574651 TI - A doubly bridged isodicyclopentadienyl zirconium complex: bis[N-(3,5 dimethylphenyl)-N-[(eta5-isodicyclopentadien-2-yl)dimethylsilyl]amido kappaN]zirconium(II) diethyl ether solvate. AB - Transmetallation of the dilithium salt of (3,5 dimethyphenylamino)(isodicyclopentadienyl)dimethylsilane by treatment with zirconium tetrachloride in a 2:1 ratio leads to the substitution of all four chloride ligands. With the applied stoichiometry, the title complex, [Zr(C(20)H(25)NSi)(2)].C(4)H(10)O, was obtained and crystallized from diethyl ether. X-ray diffraction characterization showed that both isodicyclopentadienyl ligands (alternatively called 4,5,6,7-tetrahydro-4,7-methano-1H-indene) are complexed to the metal on their exo face in a completely stereoselective manner and that they are eta(5)-bonded to the Zr atom. PMID- 12574652 TI - Bis([phthalocyaninato(2-)]arsenic(III)) tetradecaiodotetra-arsenic(III). AB - Crystals of the novel title arsenic(III) phthalocyanine complex, [As(C(32)H(16)N(8))](2)[As(4)I(14)] or [(AsPc)(+)](2).[As(4)I(14)](2-), where Pc is phthalocyaninate(2-), have been obtained by the reaction of pure powdered As with phthalonitrile under a stream of iodine vapour at 493 K. The crystals are built up of separate but interacting [AsPc](+) cations and [As(4)I(12)](2-) anions. The As atom of the [AsPc](+) unit is bonded to the four isoindole N atoms of the Pc macrocycle and lies 0.743 (2) A out of the plane defined by these four N atoms. The anionic part of the complex consists of AsI(3) and [AsI(4)](-) units joined together into an [As(4)I(14)](2-) anion. The arrangement of the oppositely charged moieties, [AsPc](+) and [As(4)I(14)](2-), in the crystal is determined mainly by ionic attraction and by donor-acceptor interactions between the [AsPc](+) and [As(4)I(14)](2-) ions. PMID- 12574653 TI - Two new heterocyclic [3.3.3.01,5]-propellanoid compounds: 2,8 dioxatricyclo[3.3.3.01,5]undecane-3,7-dione and a related dimer. AB - 2,8-dioxatricyclo[3.3.3.0(1,5)]undecane-3,7-dione, C(9)H(10)O(4), (I), is the dilactone acylal of cyclopentanone-2,2-diacetic acid. Both molecules in the asymmetric unit have conformational chirality and differ principally in the flexing of the carbon ring, which produces a resolvable conformational disorder in one of the molecules. Three intermolecular C-H...O close contacts exist. 7,7' Oxybis(2,8-dioxatricyclo[3.3.3.0(1,5)]undecan-3-one), C(18)H(22)O(7), (II), a racemate, lies on a C(2) axis and is a non-meso furanosyl furanoside dimer derived from the monoacid monoaldehyde corresponding to (I). One intermolecular C H...O close contact exists. Diminished intramolecular void space in these small propellanoids generates unusually high crystal density in both species, particularly (I). PMID- 12574654 TI - Conformation of N-methyl-4-piperidyl 2,4-dinitrobenzoate. AB - The crystal structure of N-methyl-4-piperidyl 2,4-dinitrobenzoate, C(13)H(15)N(3)O(6), (I), at 130 (2) K reveals that, in the solid state, the molecule exists in the equatorial conformation, (Ieq). Thus, the through-bond interaction present in the axial conformation, (Iax), is not strong enough to overcome the syn-diaxial interactions between the axial methyl substituent and the axial H atoms on the two piperidyl ring C atoms either side of the ester linked ring C atom. The carboxylate group in (I) is orthogonal to the aromatic ring, in contrast with other 2,4-dinitrobenzoates, which are coplanar. The piperidyl-ester C-O bond distance is 1.467 (3) A, which is actually shorter than other equatorial cyclohexyl-ester C-O distances. This shorter piperidyl-ester C-O bond distance is due to the reduced electron demand of the orthogonal ester group. PMID- 12574655 TI - The solid dihydrate of N,N-dimethyl-n-tetradecylamine oxide. AB - Crystalline N,N-dimethyl-n-tetradecylamine oxide has been prepared by reaction of liquid N,N-dimethyl-n-tetradecylamine with 70% H(2)O(2) in the presence of CO(2) as catalyst. The resulting soft low-melting solid was crystallized as the dihydrate, viz. C(16)H(35)NO.2H(2)O. The extended hydrocarbon chains pack in a parallel fashion, with the N-oxide ends of the molecules forming hydrogen bonds with the water molecules in hydrophilic layers. The N-O distance is 1.411 (3) A. PMID- 12574656 TI - Secondary interactions in 1-iodo-3-nitrobenzene and 1-iodo-3,5-dinitrobenzene. AB - The crystal packing of 1-iodo-3-nitrobenzene, C(6)H(4)INO(2), is formed by planar molecules which are linked by I...I and NO(2)...NO(2) interactions. In the case of 1-iodo-3,5-dinitrobenzene, C(6)H(3)IN(2)O(4), the NO(2) groups are not exactly coplanar with the benzene ring and the molecules form sheets linked by NO(2)...NO(2) interactions. In contrast with 4-iodonitrobenzene, the crystal structures of both title compounds do not form highly symmetrical I...NO(2) intermolecular interactions. PMID- 12574657 TI - Bis[14beta-hydroxy-3beta-O-(L-thevetosyl)-5beta-card-20(22)-enolide] methanol solvate monohydrate and 3beta-O-(L-2'-o-acetylthevetosyl)-14beta-hydroxy-5beta card-20(22)-enolide. AB - The title compounds, 2C(30)H(46)O(8).CH(3)OH.H(2)O, (I), and C(32)H(48)O(9), (II), respectively, are cardenolide glycosides which were isolated from the seeds of Cerbera odollam. There are two crystallographically independent cardenolide molecules in (I), together with one methanol and one water solvate molecule. In both (I) and (II), the steroid nuclei are in cis/trans/cis configurations, with the cyclopentane rings showing conformational flexibility, viz. an envelope conformation in (I) and a twisted conformation in (II). In both compounds, the lactone ring is nearly orthogonal to the cyclopentane ring. The packing of (I) is composed of molecular layers stabilized by five O-H...O hydrogen bonds. In the packing of (II), the molecules are packed into columns by one O-H...O hydrogen bond, and are further interconnected into a three-dimensional network by one O H...O and three C-H...O interactions. PMID- 12574658 TI - Non-mutagenic organic pigment intermediates. I. 3,3'-dipropoxybenzidine. AB - Crystals of the title compound, C(18)H(24)N(2)O(2), were grown from ethanol by slow evaporation and the structure has been determined. The molecule resides on a crystallographic inversion center and the biphenyl moiety is essentially planar. The structure forms an infinite two-dimensional array of N-H.pi(arene) interactions parallel to the (101) direction. PMID- 12574659 TI - 9,12-diiodo-1,2-dicarba-closo-dodecaborane(12). AB - The title compound, C(2)H(10)B(10)I(2), has a pseudo-icosahedral cluster geometry. The crystal structure features an intermolecular C-H...I-B hydrogen bond with a normalized H...I distance of 3.00 A. PMID- 12574660 TI - N-(1-naphthylacetyl)glycine phenacyl ester and phenacyl (1 naphthylacetoxy)acetate. AB - The structures of the bichromophoric compounds N-(1-naphthylacetyl)glycine phenacyl ester, C(22)H(19)NO(4), (I), and its oxygen analogue, phenacyl (1 naphthylacetoxy)acetate, C(22)H(18)O(5), (II), have been determined. The molecules of (I) are held together by intermolecular N-H...O hydrogen bonds between the carbonyl and N-H groups, while compound (II) does not show any hydrogen bonding in the crystal. PMID- 12574661 TI - A new estra-1,3,5(10)-triene-3,17beta-diol solvate: estradiol-methanol-water (3/2/1). AB - The title solvate of the steroid 17beta-estradiol (E(2)) with methanol and water, C(18)H(24)O(2).0.67CH(4)O.0.33H(2)O, is the first E(2) derivative to contain three crystallographically independent molecules in the asymmetric unit. The three steroid molecules, along with two methanol molecules and a water molecule, create a three-dimensional hydrogen-bonded system. Three-sided columns are formed, with the estradiol molecules aligned lengthwise parallel to (101), and joined by solvent molecules at both hydrophilic ends. The three estradiol molecules differ slightly in their ring-bowing angles, i.e. the angle between the mean plane of the A ring and that of the BCD ring; this angle ranges from 7.1 to 12.2 degrees. PMID- 12574662 TI - N-benzoylthiourea. AB - In the title compound, C(8)H(8)N(2)OS, strong intramolecular N-H.O hydrogen bonds [N.O = 2.669 (3) and 2.618 (3) A] form almost planar six-membered rings and enforce the conformation of the molecule. Two kinds of intermolecular N-H.S hydrogen bonds [N.S = 3.309 (3)-3.456 (2) A] between two symmetry-independent molecules form consecutive dimers that expand in ribbons along the [100] direction. PMID- 12574663 TI - A semi-synthetic analog of the cembranoid sarcophine. AB - The title molecule, 2(R)-[(1E,3E,7S,8S,11E,13R)-13-hydroxy-4,8,12-trimethyl-7,8 epoxycyclotetradeca-1,3,11-trien-1-yl]propane-1,2-diol, C(20)H(32)O(4), is a semi synthetic analog of sarcophine, the natural cembranoid of marine origin, isolated from the soft coral Sarcophyton glaucum. The conformation of the 14-membered ring differs substantially from that of sarcophine. The two OH groups of the propane 1,2-diol moiety form an unusual weak intramolecular hydrogen bond with an O...O distance of 2.788 (2) A, and the molecules are linked into double chains by intermolecular hydrogen bonds with O...O distances of 2.772 (2) and 2.849 (2) A. PMID- 12574664 TI - The hydrogen-bonded adduct 7,16-diazonia-18-crown-6-4-aminobenzenesulfonate-water (1/2/2). AB - In the title hydrated adduct, 1,4,10,13-tetraoxa-7,16-diazoniacyclooctadecane bis(4-aminobenzenesulfonate) dihydrate, C(12)H(28)N(2)O(4)(2+).2C(6)H(6)NO(3)S( ).2H(2)O, formed between 7,16-diaza-18-crown-6 and the dihydrate of 4 aminobenzenesulfonic acid, the macrocyclic cations lie across centres of inversion in the orthorhombic space group Pbca. The anions alone form zigzag chains, and the cations and anions together form sheets that are linked via water molecules and anions to form a three-dimensional grid. PMID- 12574665 TI - 2,5-di-2-thienylthiazolo[4,5-d]thiazole. AB - The molecules of the title compound, C(12)H(6)N(2)S(4), lie on centres of symmetry. The thiophene and thiazole rings are almost planar and their planes make a dihedral angle of 1.68 (8) degrees. In the crystal structure, there is a relatively short intermolecular S...S contact distance of 3.5786 (9) A. PMID- 12574666 TI - Hydronium (cycloheptylammonio)methylene-1,1-bisphosphonate (hydronium incadronate). AB - The structure of the title compound, H(3)O(+).C(8)H(18)NO(6)P(2)(-), adopts a zwitterionic form containing an alkylammonium group, a hydronium ion, and two negatively charged phosphonate groups. The cycloheptyl side chain adopts a twist chair conformation. The crystal packing is dominated by an extensive hydrogen bonding network. PMID- 12574667 TI - Cerebrovascular disease in Italy and Europe: it is necessary to prevent a 'pandemia'. AB - In Italy and Europe, strokes are the third most common cause of death and resulting invalidity. In the ever-increasing 80-years-old-and-over population, strokes become more serious due to the clinical presentation during the acute phase and the ten times higher mortality, but also in relation to the twice as high resulting disability as for younger subjects. With the growing number of ailing and not-self-sufficient elderly, other resources will have to be relocated to this field of public health. Then, the dependence index and the ensuing equivalence based on estimates for the first decades of 2000 will create more difficulties in retrieving the funds for social policies. However, stroke prevention is possible both through correct behavioural habits and pharmacological means. Besides the well-known preventive effects of an adequate antihypertensive, antidiabetic and/or antiaggregant/anticoagulant therapy, there is increasing evidence of the effectiveness of statin therapy in stroke prevention. Subjects with a personal history of cerebrovascular events have an increased coronary risk and vice versa. The greatest part of the risk factors for the cerebrovascular disease coincides with those for cardiovascular disease, for which the correction of the former automatically involves a reduction in incidence of both pathologies. In this context, a statin's rational use can therefore represent an important tool for the combined prevention of the two pathologies. Finally, different hypotheses link the origin of Alzheimer's disease to that of progressive cerebrovascular dementia caused by cerebral microcirculation damage. The aim of this review is to resume the actual knowledge about the epidemiology of cerebrovascular disease in Italy and Europe, and about the means available to prevent this phenomenon. PMID- 12574668 TI - Age-related changes in catecholaminergic nerve fibers of rat heart and coronary vessels. AB - BACKGROUND: Age-related changes of catecholaminergic nerve fibers supplying the myocardium and the coronary vessels were studied in adult and in old rats. OBJECTIVE: The purpose of this study was to investigate whether there are quantitative changes in myocardial and/or coronary catecholaminergic nerve fibers and related neurotransmitters. METHODS: Samples of myocardium and/or coronary vessels were studied by means of glyoxylic acid induced fluorescence, aldehyde induced fluorescence, and neuropeptide Y immunoreactivity. All morphological results were submitted to quantitative analysis of images. Biochemical dosage of proteins and of norepinephrine was also performed, and morphological data were compared with biochemical ones. Finally, all data were subjected to statistical analysis. RESULTS: Our findings are indicative of an age-related decrease of all types of fluorescent sympathetic nerve fibers in rat heart and coronary arteries. Moreover, our data show that aging induces a strong decrease of norepinephrine levels in tissue homogenates of rat heart and coronary vessels. The majority of the catecholaminergic nerve fibers that show strong age-related changes are not contiguous with contractile cells of myocardium and/or of vascular wall. In fact, these fibers are located in adventitial (or epicardial) and in endothelial (or endocardial) layers. CONCLUSION: The decrease of the catecholaminergic innervation of heart and coronary vessels of old rats can be in close relationships to the poor cardiac and/or coronary functions observed in old animals. PMID- 12574669 TI - Short-term effects of improved glycemic control on cognitive function in patients with type 2 diabetes. AB - BACKGROUND: According to numerous studies, type 2 diabetes is associated with mild cognitive dysfunction, and there is some evidence suggesting favorable effects of improved metabolic control on the mental capability of elderly diabetic patients. OBJECTIVE: To compare patients with type 2 diabetes to normal controls with respect to cognitive performance and to investigate the consequences of glycemic adjustment. METHODS: 53 patients with type 2 diabetes, most of them in secondary failure on oral antidiabetic drugs, but free from conditions which may cause brain dysfunction, were included (mean age 58.8 +/- 6.1 years, duration of disease 12.0 +/- 6.4 years). They were examined prior to (t1), and following (t2) glycemic adjustment with a time interval of approximately 2 weeks. 29 non-diabetic controls, comparable with regard to age, gender, education and verbal intelligence were examined twice with a corresponding time interval. Cognitive performance was assessed by well standardized tests with a focus on attention/concentration, psychomotor speed, verbal fluency and verbal memory; mood status by two self-rating scales. Restoration of glycemic control included insulin treatment in the majority of patients (46/53). RESULTS: Diabetic subjects scored significantly lower in all cognitive tests used, while they did not differ from controls in mood status. From t1 to t2 they improved in those tests measuring attention/concentration, and psychomotor speed. With regard to similar changes in controls, we interpret these improvements as practice effects rather than the consequence of altered metabolic control. CONCLUSION: In a sample of patients with long-standing type 2 diabetes we could not confirm previous reports of improved cognitive capacity with restoration of glycemic control. Further studies on the effects of changes in control of blood glucose on cognitive performance in type 2 diabetes should be conducted with special regard to drugs used to lower blood glucose. PMID- 12574670 TI - Evidence-based guidelines for the secondary prevention of falls in older adults. AB - BACKGROUND: Falls are a significant problem for older adults. Individuals who have sustained a fall come to the attention of health care providers and are at risk of further falls. To promote the highest quality of care and reduce variation in care, a practice guideline is needed. Summarization of evidence regarding falls may be useful to researchers in this field. OBJECTIVES: To provide evidence-based guidelines of assessment and treatment to prevent falls in older adults and to provide researchers with tables of risk factor studies and randomized controlled trials of falls prevention. METHODS: A template for the development of practice guidelines from the Agency for Health Care Policy and Research was used. Evidence for risk factors was accepted from prospective studies with more than 80% follow-up. Potentially modifiable risk factors were selected and a schema for evaluating the importance of each risk factor was used. Evidence for interventions was examined from randomized controlled trials and strength of the evidence was graded. Recommendations for aspects of care where judgment was required were made by panel consensus. RESULTS: Information was drawn from 46 risk factor studies and 37 randomized controlled trials to develop a practice guideline consisting of assessment items and recommended interventions for community-dwelling and institution-dwelling older adults separately. For clinicians, a check list is provided. Summary tables of the results of studies are given to substantiate the recommendations. CONCLUSIONS: For community dwelling older adults, there is strong evidence for multi-factorial specific risk assessment and targeted treatment. Balance exercises are recommended for all individuals who have had a fall and there is evidence for a program of home physiotherapy for women over 80 years of age regardless of risk factor status. For institutional settings, the establishment of a falls program for safety checks, ongoing staff education and monitoring is substantiated by research. Residents who have fallen need to be assessed for specific risk factors and clinical indicators to determine relevant management options. PMID- 12574671 TI - Iron and ascorbic acid concentrations in human dermis with regard to age and body sites. AB - BACKGROUND: Reactive oxygen species (ROS) contribute to processes relating to cutaneous aging. Iron catalyses ROS formation whereas ascorbic acid (AA) plays a fundamental role in defending the organism against undesirable ROS action. OBJECTIVE: The aim of this work was to determine the ex vivo iron and AA concentrations in human dermis from different age groups to better understand their role. METHODS: Skin fragments were collected from 66 female patients during surgical operations and were grouped according to age: group I (<15 years, before puberty, n = 12), group II (15-50 years, adults, n = 42), and group III (>50 years, advanced age adults, n = 12). Two sites were investigated: the abdomen (unexposed areas) and face (exposed sites). Iron and AA were collected from human dermis by microdialysis and assessed by atomic absorption spectrometry and gas chromatography mass spectrometry, respectively. RESULTS: Iron concentrations in the dermis were significantly higher in group III (27.4 +/- 20.9 microg/l) than in group I (13.8 +/- 3.3 microg/l; p< 0.05 ). An inverse correlation between AA dermis levels and increasing age was detected. For groups III and I, iron and AA concentrations were significantly different in dermis from the face compared to that of the abdomen (p < 0.05). CONCLUSION: This study shows for the first time that there is a direct relationship between iron and AA concentrations in the dermis and aging. Moreover, iron and AA concentrations differed according to body site. PMID- 12574672 TI - New frontiers in the future of aging: from successful aging of the young old to the dilemmas of the fourth age. AB - We review research findings on the oldest old that demonstrate that the fourth age entails a level of biocultural incompleteness, vulnerability and unpredictability that is distinct from the positive views of the third age (young old). The oldest old are at the limits of their functional capacity and science and social policy are constrained in terms of intervention. New theoretical and practical endeavors are required to deal with the challenges of increased numbers of the oldest old and the associated prevalence of frailty and forms of psychological mortality (e.g., loss of identity, psychological autonomy and a sense of control). Investigation of the fourth age is a new and challenging interdisciplinary research territory. Future study and discussion should focus on the critical question of whether the continuing major investments into extending the life span into the fourth age actually reduce the opportunities of an increasing number of people to live and die in dignity. PMID- 12574673 TI - Your report card is coming-are you ready? PMID- 12574674 TI - Ethical dilemma: a mother's orders about truth telling. PMID- 12574675 TI - Home Care LPN Utilization. PMID- 12574676 TI - It's a leaner, meaner year: what's new in the 2003 Joint Commission home care and hospice standards: part I. PMID- 12574677 TI - Coumadin knowledge deficits: do recently hospitalized patients know how to safely manage the medication? AB - There is much that patients must know to be able to safely manage warfarin (Coumadin). This study examined the extent of Coumadin-related knowledge patients living at home had 1 week following hospital discharge. Findings indicate that home care nurses need to concentrate on supplementing the inadequate knowledge patients have about Coumadin therapy, especially among the elderly. PMID- 12574678 TI - Understanding benchmarks. PMID- 12574679 TI - Connecting with older clients. PMID- 12574680 TI - Central venous access device occlusions: part 1: thrombotic causes and treatment. AB - Many home care patients have central venous access devices to administer a variety of infusion therapies. Catheter occlusion is a potential complication. This month, catheter occlusion due to thrombotic causes is addressed. In many cases, thrombotic occlusions can be treated, sparing patients trauma and the expense of replacing the catheter. Thrombolytic drug therapy using the drug alteplase can be performed in the home setting to "declot" central venous access devices. Next month, occlusion due to nonthrombotic causes will be described. PMID- 12574681 TI - Medicare reform for prescription drugs. PMID- 12574682 TI - Frequently asked questions about computer technology: information for managers. PMID- 12574683 TI - Customer satisfaction and workplace safety. PMID- 12574685 TI - Smallpox and smallpox vaccine--just the facts. PMID- 12574686 TI - Assuring the right provider for the right patient at the right time. PMID- 12574688 TI - Undiagnosed airflow obstruction: prevalence and implications. AB - Conditions associated with airflow obstruction are often over- and underdiagnosed. Prevalence estimates of undiagnosed airflow obstruction (UDAO) range from 3 to 12%. UDAO is a nonspecific physiologic abnormality that may be caused by a number of factors (eg, cigarette smoking) and can be the manifestation of many different disorders. The higher occurrence of UDAO among men, current or former smokers, and with advancing age provide preliminary evidence on probable causes and diagnoses (ie, chronic obstructive pulmonary disease). While cigarette smoking is associated with UDAO, a substantial proportion of persons have never smoked, particularly among women. Few studies suggest that this condition is associated with increased morbidity and mortality. While there is currently no evidence to support screening for UDAO, case-finding may have a role among persons with respiratory symptoms, who have ever smoked, with a family history of respiratory disease, or with occupational exposures to dusts or fumes. PMID- 12574689 TI - Chemokines and their receptors in asthma and chronic obstructive pulmonary disease. AB - Recruitment of inflammatory cells into the airways is a critical event that triggers and sustains the clinical manifestations of asthma and chronic obstructive pulmonary disease. Since the identification of the family of chemotactic cytokines, known as chemokines, as critical regulators of cell trafficking in the immune system, these molecules have come to the center stage in the field of inflammation and immunity. The goal of this article will be to summarize the recent developments in our understanding of the complex role that chemokines play in the pathogenesis of asthma and COPD. PMID- 12574690 TI - Viral infections in obstructive airway diseases. AB - The most common syndromes associated with obstructive lung disease are asthma and chronic obstructive pulmonary disease (COPD). Evidence for a viral etiology of asthmatic exacerbations is well known, but evidence for a role for viruses in COPD exacerbation is just emerging. Viruses may cause chronic infection in both diseases. This paper reviews some studies on the effects of respiratory viruses on asthma and COPD published in 2002 and discusses their relevance to current thinking in pulmonary medicine. PMID- 12574691 TI - Evidence-based approach to acute exacerbations of COPD. AB - Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the United States, and it accounts for approximately 500,000 hospitalizations for exacerbations each year. New definitions of acute COPD exacerbation have been suggested, but the one used by Anthonisen et al. is still widely accepted. It requires the presence of one or more of the following findings: increase in sputum purulence, increase in sputum volume, and worsening of dyspnea. Patients with COPD typically present with acute decompensation of their disease one to three times a year, and 3% to 16% of these will require hospital admission. Hospital mortality of these admissions ranges from 3% to 10% in severe COPD patients, and it is much higher for patients requiring ICU admission. The etiology of the exacerbations is mainly infectious (up to 80%). Other conditions such as heart failure, pulmonary embolism, nonpulmonary infections, and pneumothorax can mimic an acute exacerbation or possibly act as "triggers." Baseline chest radiography and arterial blood gas analysis during an exacerbation are recommended. Oxygen administration through a venturi mask seems to be appropriate and safe, and the oxygen saturation should be kept just above 90%. Either a short acting beta 2-agonist or an anticholinergic is the preferred bronchodilator agent. The choice between the two depends largely on potential undesirable side effects and the patient's coexistent conditions. Adding a second bronchodilator to the first one does not seem to offer much benefit. The evidence suggests similar benefit of MDIs when compared with nebulized treatment for bronchodilator delivery. If MDIs are to be used, spacer devices are recommended. Steroids do improve several outcomes during an acute COPD exacerbation, and a 10- to 14-day course seems appropriate. Antibiotic use has been shown to be beneficial, especially for patients with severe exacerbation. Changes in bacteria strains have been documented during exacerbations, and newer generations of antibiotics might offer a better response rate. There is no role for mucolytic agents or chest physiotherapy in the acute exacerbation setting. Noninvasive positive pressure ventilation might benefit a group of patients with rapid decline in respiratory function and gas exchange. It has the potential to decrease the need for intubation and invasive mechanical ventilation and possibly decrease in-hospital mortality. PMID- 12574692 TI - Approaches to slowing the progression of COPD. AB - Chronic obstructive pulmonary disease (COPD) is a major health problem across the world and its medical, societal and economic impacts continue to grow. There are several hypotheses regarding the pathogenesis of COPD and important new information on airway inflammation, oxidative stress and proteolysis in the lungs that are important for the development of effective treatments. The differences in susceptibility to COPD among cigarette smokers and the common familial risk of developing disease point to important genetic influences. The identification of candidate gene loci and various gene polymorphisms have improved our understanding of the mechanisms by which genetic factors predispose for COPD. The beneficial effects of smoking cessation in slowing the decline in lung function and the progression of disease have been clearly established. Whether other factors such as mucus hypersecretion, respiratory infections and airway hyperreactivity contribute to disease progression independent of cigarette smoking is still being debated. There is new interest in dietary strategies to prevent or control COPD and preliminary information suggesting the usefulness of diets favoring antioxidant-rich foods. On the other hand several large, long term, randomized, controlled clinical trials have failed to show a beneficial effect of either anticholinergics or corticosteroids in slowing the rate of decline in lung function in COPD. Corticosteroid treatment, however, appears to reduce the frequency of COPD exacerbations and there is some suggestion that it may also reduce the risk of death in patients with severe COPD. PMID- 12574693 TI - Pathophysiology of pulmonary hypertension due to lung disease. AB - Pulmonary hypertension (PH) often complicates the course of patients with advanced lung disease, and it is associated with a worse prognosis. Per the recent classification of pulmonary hypertensive disorders, PH due to lung disease is considered as a separate category within a group of disorders that was previously referred to as "secondary" PH. Among the lung diseases associated with PH, the incidence and clinical course of PH is best known for patients with COPD. Per studies in patients with COPD and other lung disorders, it is evident that the pathophysiology and treatment of these disorders is generally distinct from that of pulmonary arterial hypertensive disorders. Changes in the pulmonary vasculature that accompany elevations in pulmonary vascular pressure are generally referred to as pulmonary vascular remodeling. Chronic hypoxia is well known to cause pulmonary vascular remodeling and PH, and it is the major mechanism implicated for the development of PH in patients with lung disease. Other mediators have also been implicated in the pathogenesis of PH in animal models and patients with PH, including patients with pulmonary diseases. General features of pulmonary vascular remodeling are discussed with particular emphasis on those changes that have been described in patients with lung diseases. Recent discoveries in these areas are also reviewed, and findings in pulmonary arterial hypertensive diseases are contrasted with those found in patients with PH due to lung diseases. Some of these discoveries have already led to new treatment strategies for patients with the most severe forms of PH. PH due to lung diseases shares some common pathophysiologic features with pulmonary arterial hypertension. Therefore, it is likely that these discoveries and new treatments will also be extended to benefit patients with PH due to lung disease. PMID- 12574694 TI - Advances in the treatment of secondary pulmonary hypertension. AB - Pulmonary hypertension (PH) occurs frequently in parenchymal lung disease and is usually correlated with increased mortality. Thus, the treatment of PH in patients with lung disease has been an active area of interest. Secondary pulmonary hypertension (SPH), whether from parenchymal lung disease or other etiologies, is more common than primary pulmonary hypertension (PPH). In 2002, two new medications, oral bosentan and subcutaneous treprostinil, were released for the treatment of pulmonary arterial hypertension (PAH). These new agents are not restricted to use in PPH, as they are approved for use in PAH in general. It is reasonable to consider the use of these medications in select patients with SPH caused by parenchymal lung disease, although these groups have not yet been studied in clinical trials. The initial hemodynamic evaluation of SPH patients, the potential use of these new medications in the context of standard care, and the assessment of response to therapy are discussed in this update. A relevant case report is used to illustrate use of these new agents in SPH, and ongoing clinical trials are reviewed. The available treatment options for patients with SPH are rapidly improving. PMID- 12574695 TI - Effects of ozone on lung function and lung diseases. AB - Ozone (O3) is an air pollutant produced by sunlight-driven reactions involving the oxides of nitrogen and volatile organic compounds. The population of many large metropolitan areas in the US is exposed to high levels of O3, particularly in the summer months. Individuals exposed to O3 levels in human experiments at higher than common ambient levels develop reversible reductions in lung function often associated with symptoms, such as airway hyperreactivity and lung inflammation. Animal models have helped characterize potential mechanisms of lung injury from O3 exposure. Defining the adverse effects of chronic exposure to ambient levels of O3 on lung function and disease have been challenging, in part due to the presence of co-pollutants, such as particulate matter. The US Environmental Protection Agency's 1997 revised standard for O3 (0.08 ppm averaged over 8 hours) is designed to provide better protection to susceptible individuals. The revised standard is being implemented following the failure of court challenges. PMID- 12574696 TI - Respiratory effects of long-term exposure to cotton dust. AB - Traditionally, the concept of cotton dust-related disease focused on acute airway responses. A possible connection of long-term exposure to cotton dust with chronic obstructive airway disease was not addressed until the past two decades. This review summarizes the latest findings relevant to this topic that were observed from longitudinal cohort studies and discusses the relation between acute airway responses and chronic losses of lung function. PMID- 12574698 TI - Depression in patients with heart failure: physiologic effects, incidence, and relation to mortality. AB - Heart failure affects 4.8 million people in the United States. Patients depressed after myocardial infarction have increased morbidity and mortality. Only a few studies have investigated the effects of depression in patients with heart failure. The incidence of depression in heart failure ranges from 13% to 77.5%. Men with heart failure are more likely to become depressed than the general population. Depression incidence is higher in hospitalized patients with heart failure than in stabilized outpatients. In patients with heart failure, depression is associated with mortality. Physiologic changes, which occur in depressed patients, have been implicated as possibly contributing to the increased mortality. Nurses have a major role in the management of patients with heart failure and can be pivotal in the detection and treatment of depression in these patients. Reduction in depression is likely to decrease morality in patients with heart failure. PMID- 12574699 TI - Addressing current challenges to cardiac rehabilitation care. AB - Cardiac rehabilitation, the structured programming of exercise and risk reduction teaching and counseling to promote healthy living with heart disease, is grounded in a strong nurse-patient relationship. This relationship provides a foundation for meeting current healthcare challenges. Three challenges are presented and addressed through specific examples. The challenge of the changing cardiac rehabilitation population is addressed through the example of the heart failure patient. The challenge to create innovative programming is addressed through the example of the Therapeutic Rehabilitative Intensive Program, developed to meet the needs of people who live in communities without cardiac rehabilitation services. The challenge to measure outcomes is addressed through the example of choosing a blood pressure measurement method. In addressing each challenge, the role of the advanced practice nurse is described. PMID- 12574700 TI - Psychoneuroimmunology in critically ill patients. AB - Psychoneuroimmunology is the study of the interactions among behavior, neural, and endocrine functions and the immune system. The purpose of this review is to briefly summarize the evidence concerning interactions among behavior, the neuroendocrine system, and the immune system, and to show how this evidence relates to critical care patients. It has been shown that the immune function of many patients in the intensive care unit is suppressed as a result of trauma, sepsis, or profound physiologic and psychological stress. Three of the most common stressors among patients in the intensive care unit are pain, sleep deprivation, and fear or anxiety. Findings have shown each of these stressors to be associated with decreased immune functioning. Nurses have an important responsibility to protect their patients from infection and promote their ability to heal. Several actions are suggested that can help the nurse achieve these goals. It is hoped that nurses would keep these interactions in mind while caring for their patients in the intensive care unit. PMID- 12574701 TI - Comparison of family stresses, strengths, and outcomes after trauma and surgery. AB - This study aimed to compare family stresses, strengths, and outcomes after motor vehicle crashes, gunshot wounds, and coronary artery bypass graft surgery. A multivariate descriptive design based on the resiliency model of family stress was used. A convenience sample of 127 family members participated within 2 days of admission to the intensive care unit. Family stresses were measured using the Family Inventory of Life Events and the Acute Physiology, Age, and Chronic Health Evaluation Scale (APACHE III). Family strengths were measured by the Family Hardiness Index and the Family Crisis Oriented Personal Evaluation Scale. Family outcomes were measured by the Family Member Well-Being Index and the Family Adaptation Scale. Family members ranged in age from 18 to 80 years (Mean, 42 years). Most of the participants in the study were women (64%) with previous intensive care unit experience (83%). The patients ranged in age from 19 to 78 years (Mean, 44 years). Multivariate analysis of variance was used to control for group differences in family member age, education, and income along with patient age and race. The results indicated that family members of patients who have experienced motor vehicle crashes, gunshot wounds, or coronary artery bypass graft surgery reported no significant differences in family strengths of hardiness and family outcomes of well-being and adaptation. However, families of patients with gunshot wounds reported significantly more stress (F = 7.94; P< 0.01) and less use of coping strategies (F = 4.33; P< 0.01) than families of patients who have experienced motor vehicle crashes or coronary artery bypass graft surgery. Interventions that address family stress and develop or mobilize coping are needed for selected families after admission of a family member to the intensive care unit. PMID- 12574702 TI - Quality of life experienced by adults with congenital heart disease. AB - Adults with congenital heart disease (CHD) represent a growing population of patients. Medical and surgical advances have increased the number of CHD adult survivors, which may create quality-of-life (QOL) issues not previously considered. Quality-of-life issues pertinent to this patient population involve health and life insurance acquisition, birth control, genetic counseling, pregnancy concerns, employment, and independent living arrangements. The purpose of this study was to describe the QOL of adults with CHD. The study used a prospective cross-sectional case-control design to examine QOL using the Sickness Impact Profile (SIP). The study participants were a sample of 124 adults with CHD from an outpatient cardiology clinic in a metropolitan university-affiliated teaching hospital in the Northeast and 124 matched healthy control subjects. Between the participants and the matched control subjects, there was a significant difference in the total mean SIP score, the physical and psychosocial dimension scores, and all the category scores (P < 0.05). The areas of life the adults with CHD reported as lacking in quality involved the categories of work (SIP of 11.1, moderate disability) and sleep and rest (SIP of 9.03, mild disability). The results of this study indicate that the SIP can be used for quantitative and subjective QOL assessment of adults with CHD. It is suggested that cardiac advanced practice nurses use the results of this study to develop appropriate information, counseling, and anticipatory guidance for this patient population. PMID- 12574703 TI - Quality of life experienced by severely injured trauma survivors. AB - Advances in technology and the expansion of the emergency medical system have made emergency care available to large numbers of people experiencing trauma. Assessing the quality of life experienced by trauma survivors may assist in the development of interventions to optimize the outcomes in this patient population. The purpose of this study was to assess the quality of life experienced by severely injured trauma survivors, and to determine if a relation exists between the severity of the injury and the quality of life, the impact on the family of survivors, and the community resources needed by the survivors. Severely injured survivors of trauma in this study were defined as patients with a severity of injury severity score (ISS) of 25 or more and a trauma injury score (TRISS) of 0.90 or less. In this descriptive study, telephone interviews were conducted to administer the Sickness Impact Profile (SIP), an analog scale, and a demographic data form. All the survivors were interviewed at least 6 months after discharge from the hospital. The data collected on 64 survivors showed that they experienced a significantly poorer quality of life after the trauma incidents, as indicated by both the SIP scores and the analog scores. Although most of the SIP's 12 behavior categories indicated severe disruption, the behavior categories of work, recreation and pastimes, home management, and sleep and rest were the most negatively affected. Patients and families should be prepared for the physical, emotional, and financial disruptions that occur after severe traumatic injury. Support services, including community resources, are needed to optimize outcomes after discharge. PMID- 12574704 TI - Quality of life among patients with a left ventricular assist device: what is new? AB - As more patients have a diagnosis of end-stage heart failure, nurses are more likely to encounter the use of left ventricular assist devices (LVADs). Currently, LVADs are used as a bridge to cardiac transplantation. However, current research suggests that LVADs may be used as destination therapy for those unable to undergo cardiac transplantation. The technology has been developed to sustain life, but what is the impact of technology on the quality of life? This article reviews current research on the quality of life with an LVAD. PMID- 12574705 TI - Clinical management of stressors perceived by patients on mechanical ventilation. AB - Psychological and psychosocial stressors perceived by the mechanically ventilated patient include intensive care unit environmental factors, communication factors, stressful symptoms, and the effectiveness of interventions. The studies reviewed in this article showed four stressors commonly identified by mechanically ventilated patients including dyspnea, anxiety, fear, and pain. Few interventional studies to reduce these stressors are available in the literature. Four interventions including hypnosis and relaxation, patient education and information sharing, music therapy, and supportive touch have been investigated in the literature and may be helpful in reducing patient stress. The advanced practice nurse is instrumental in the assessment of patient-perceived stressors while on the ventilator, and in the planning and implementation of appropriate interventions to reduce stressors and facilitate optimal ventilation, weaning, or both. PMID- 12574706 TI - Effect of a treatment interference protocol on clinical decision making for restraint use in the intensive care unit: a pilot study. AB - The literature is replete with articles describing restraint reduction strategies used in long-term care settings, geriatric specialty units, and medical/surgical units in the acute care setting. The feasibility, effectiveness, and appropriateness of such strategies cannot be capriciously applied to the intensive care setting. This article provides an overview of the implementation and outcomes of a pilot study using an algorithmic approach that is clinically appropriate and justifiable for restraint use in the intensive care environment. It provides the critical care nurse with a standardized method for decision analysis when managing patients at risk for treatment interference. PMID- 12574707 TI - Identification and management of delirium in the critically ill patient with cancer. AB - Rather than a specific entity, delirium is at the midpoint on a spectrum of potential mental status changes that ranges from full consciousness to deep coma. The extremes are relatively easy to recognize, but other points along the spectrum may go unrecognized or be misdiagnosed. If recognized and treated expeditiously, delirium may be reversed in some patients. It is imperative that those caring for critically ill patients with cancer have the knowledge and tools necessary to identify and manage delirium appropriately. Although all critically ill patients are at risk for delirium, cancer presents additional assaults to the central nervous system via direct tumor invasion or iatrogenic provocations. This article describes delirium in cancer, and addresses diagnostic and management issues across the course of the disease. PMID- 12574708 TI - Intimate examinations are carried out without consent. PMID- 12574709 TI - The NHS University should benefit all staff in the NHS. PMID- 12574710 TI - Nurse who covered up for a sister who was having problems. PMID- 12574711 TI - The male menopause: possible causes, symptoms and treatment. AB - This article discusses the controversial concept of the male menopause, by explaining what the literature reveals about this condition. It does not wish to put forward a case for treating men with the male menopause with hormone replacement therapy, merely to provide information to nurses who may nurse men with the symptoms associated with the disorder. Definitions of key terms, i.e. male menopause, are considered and the endocrinological changes that occur with ageing are outlined. Assessment of the extent of the problem is confounded by the concept of definition. Some middle-aged men often describe similar symptoms that women describe associated with the menopause; therefore it may be reasonable to suggest that an andropause with an endocrine basis does exist. The investigations undertaken to make a diagnosis are described and treatments available are discussed. A shared approach to the care for men with menopausal symptoms is advocated. The role the nurse plays is described and recommendations for practice suggested. Finally, a conclusion is offered and a call is made for further empirical investigations to be made available, to ensure that men are provided with safe effective care based on the best available evidence. PMID- 12574712 TI - A model for patient-centred nurse consulting in primary care. AB - A model for patient-centred nurse consulting, developed in the context of secondary prevention of cardiovascular disease, is reported in this article. The model was created in the absence of any existing model for nurse consulting. Heart disease clinics are increasingly being used to implement the National Service Framework for Coronary Heart Disease and it is anticipated that this model will be of practical use to nurses running these clinics. The model consists of eight steps, broadly following the chronology of the consultation. Each step contains a question for the nurse to ask him/herself and a task to complete. The model was developed as part of a research project on the influence of customized computer software used on nurse consultations with patients with heart disease. PMID- 12574713 TI - Research endeavours into suicide: a need to shift the emphasis. AB - According to the World Health Organization (WHO, 2002), approximately one million people died as a result of suicide in the year 2000. Perhaps more worrisome, evidence provided by the WHO (2002) indicates that the global rate of suicide has continued to rise since 1950. Consequently, suicide should be regarded as a global problem and one that is increasing in magnitude. Drawing predominantly on policy and empirical literature emanating from Canada and the UK, this article will show that, despite a substantial research effort and the production of an associated literature, suicide rates continue to rise in the example countries. Even given the existence of numerous positivistically oriented studies, and the introduction of a range of strategies to help prevent suicide, significant reductions in suicide rates have not been achieved. Similarly, while there exists a substantial literature on the issue of suicide, there are many gaps in our knowledge and our understanding of the experiences, and the meanings attributed to these experiences that motivate people to attempt suicide is far from complete. Accordingly, the author argues that there is an urgent need to better understand the particular life experiences and the meanings that individuals attach to suicidal experiences. In order to design interventions to help reduce the suicide rate, whether these are interventions at the pre-primary, primary or secondary level of care, it is argued that it is necessary to gain a more detailed and comprehensive understanding of this highly complex behaviour. Consequently, this article makes the case for the use of hermeneutic, phenomenological investigations, in order to further elucidate the lived experiences of people who have attempted suicide. PMID- 12574714 TI - Violence against nurses in hospitals: prevalence and effects. AB - There is abundant evidence to suggest that violence in healthcare settings, especially in emergency departments (EDs) and psychiatry, is escalating and that nurses are particularly vulnerable. The authors, therefore, investigated the prevalence and effects of violence against nurses in an ED in a general hospital setting in Kuwait. A 12-item frequency-weighted questionnaire was used to measure rates, frequency and severity of violence. The questions related to the experience of violent incidents during the past year. Seventy out of 81 nurses experienced verbal insults or threats of imminent violence and 13 were also physically attacked during the 1-year period. Sixty-seven out of 70 nurses suffered from one or more after-effects, including flashbacks, sleeplessness, fearfulness, depression or taking time off work. Violence in healthcare services often reflects the community in which service is provided. Our findings suggest that doctors experience more violence but nurses suffer from more after-effects of violence at work. PMID- 12574715 TI - Laws relating to HIV/AIDS infection and the rights of employees. AB - Following a holiday in Africa, Brenda, a nurse, was told that she had probably been infected with the human immunodeficiency virus. She felt reasonably fit and since she worked in ophthalmic outpatients, an area where there was a very low risk of infecting patients, she wanted to keep this information to herself. What is the law? PMID- 12574716 TI - Autonomy, privacy and informed consent 2: postnatal perspective. AB - The nursing and healthcare ethics literature over the past 10 years has focused on issues of patient autonomy and patient rights. Despite the growing volume of literature exploring such topics, there is little empirical work investigating what is actually happening in clinical nursing or midwifery practice in relation to patient autonomy, privacy or informed consent, from the perspective of either patients or staff. This four-part series reports the results of a Scottish study that formed part of a multisite comparative research project funded by the European Commission, investigating issues of patient autonomy, privacy and informed consent. This article, the second of four, explores the issues of autonomy, privacy and informed consent in maternity care. The research questions asked were: (1) What is the perception of mothers' autonomy, privacy and informed consent in Scottish NHS hospitals, from the point of view of both mothers and midwives? (2) Are there differences in the perceptions of mothers and midwives on these issues? Data were collected by a self-completion questionnaire for mothers (n = 243) and staff (n = 170) on postnatal units in both district general and university teaching hospital. Results indicated that there are differences between the perceptions of mothers and midwives in relation to mothers' autonomy, privacy and informed consent. Most differences were found in the information giving and decision-making elements of autonomy. PMID- 12574717 TI - Clinical nursing skills must be recognized. PMID- 12574718 TI - Standards of infection control are declining in hospitals. PMID- 12574719 TI - Managing and improving healthcare delivery in 2003. PMID- 12574720 TI - Nurse who let herself be manipulated by a patient. PMID- 12574721 TI - Dietary needs for bone health and the prevention of osteoporosis. AB - Osteoporosis is identified as a painful, disabling and disfiguring health deficit that is entirely preventable given early detection and reversal of the causes- one of which is deficiency of nutrients required for bone health. The notable examples of nutrients where deficiency is directly related to the onset of osteoporosis are identified as calcium and vitamin D. The reasons why deficiency occurs are analysed in this article and high-risk situations for deficiency identified and discussed. The use of phytooestrogens in dietary prevention of osteoporosis is explored to include analysis of foods containing phytooestrogens, the quantities required, and product variability. Given the increasing role of phytooestrogens in dietary prevention of osteoporosis, discussion in this section looks at the need for food labelling, including phytooestrogen content, so that consumers can make reasoned choices as to the quantities they require and the dietary sources from which this will be obtained. PMID- 12574722 TI - Infection control and maintaining hygiene and cleanliness. PMID- 12574723 TI - Development of nurse specialists/advanced practitioners in Ireland. AB - An Bord Altranais (Irish Nursing Board) published "Report of the Commission on Nursing: A Blueprint for the Future" in 1998. This report was the result of collaborative work among those involved in nursing in Ireland. It recommended the establishment of clinical career pathways, which would allow a nurse with extensive experience and advanced expertise, as well as an appropriate course, to be recognized as a clinical nurse specialist. Further advancement along this pathway could possibly enable the nurse to progress to an advanced practitioner grade. The report acknowledged that there are "various and differing interpretations" of the title "practitioner". This article explores the growth and development of the roles of the clinical nurse specialist and advanced nurse practitioners in Ireland over the past number of years. The literature reflects elements of confusion that exist in some areas about the role of a clinical nurse specialist and an advanced nurse practitioner. The "Review of the Scope of Nursing and Midwifery Practice" (An Board Altranais, 2000) recognized a lack of clear differentiation between the role of the clinical nurse specialist and the advanced nurse practitioner. Overall, some clear roles are identified for the nurse who advances or wishes to advance along clinical career pathways in nursing. However, there still appears to be a certain degree of confusion about the use of titles such as "nurse specialist" and/or "nurse practitioner". The guidelines about the level of practice and education/training required for the nurse moving along the pathways of clinical nurse specialism and/or advanced nursing practice is somewhat obscure and is interpreted in different ways in the literature. PMID- 12574724 TI - Role of a paediatric nurse in primary care 2: research findings. AB - This article is the second of a two-part article that presents the findings of a study focusing on the role and function of two qualified children's nurses, each working within a different primary healthcare team (PHCT). The first part described the variation in settings for community paediatric nurses, the paucity of literature discussing the role of a paediatric nurse working as a member of a PHCT, the application of a research methodology that incorporated practitioner values and experience, the application of naturalist inquiry and the use of a formative evaluation strategy which enabled the researcher to be involved in directing and supporting the research nurses with the data collection (Vol 11(22): 1452-60). This second part focuses on the findings and discussions where it was demonstrated that qualified children's nurses working in community setting contributes to raising the overall support and quality of care for a whole range of conditions currently perceived as low profile. There was evidence that these nurses catered for unmet needs and were, at times, functioning as nurse practitioners and on occasions as advanced practitioners. These findings may contribute as a catalyst for change in informing the development of children's nursing services as part of the Children's National Service Framework. PMID- 12574725 TI - Autonomy, privacy and informed consent 1: concepts and definitions. AB - This article is the first in a four-part series that explores the concepts of patient autonomy, privacy and informed consent in the context of the provision of nursing care. In this first article an overview of the concepts is provided, and some of the difficulties with definitions of these concepts are considered. The dearth of empirical exploration of the operationalization of the concepts of patient autonomy, privacy and informed consent within nursing practice is highlighted. The second, third and fourth articles report a series of empirical studies carried out with patients and staff in postnatal care, in continuing care of elderly people and in acute surgical care within Scottish NHS settings. These studies are part of a concerted effort to address the lack of empirical investigation of the ethical dimension of nursing care provision and clinical nursing practice. PMID- 12574726 TI - Health-related quality of life tools for venous-ulcerated patients. AB - Healthcare manufacturing companies have developed many new and novel materials for wound care. They claim that the dressing materials will improve the healing progress of the wound or have a therapeutic value and have sought to assess their efficacy and effectiveness in clinical trials. These claims should be clearly supported by the perspectives of those who actually use the devices. Compression therapy, which is regarded as an effective treatment for venous leg ulcers, is far from ideal owing to a number of limitations that it possesses, such as patient discomfort, difficulty experienced in applying correct compression on the limbs, choosing a correct dressing for the wound or finding differences between the performance of various devices. It is necessary to investigate the patient's views and perspectives while randomized controlled clinical trials are conducted. In this review the concept of quality of life (QoL) and the tools used to measure QoL and studies that were conducted with patients suffering from chronic venous leg ulcers and undergoing therapy are considered, along with contributory factors to the healing process. In addition, a questionnaire modified from the original format is recommended as the suitable tool for assessing the QoL of patients suffering from venous leg ulcers while participating in clinical trials. PMID- 12574727 TI - Nurses must not integrate medical tasks into nursing. PMID- 12574728 TI - Agenda for Change, or recipe for confusion? PMID- 12574729 TI - Assessment for long-term care: a snapshot of nursing practice. AB - A study was carried out to examine nurses' assessments of clients to provide a basis for long-term placement decision-making, in both the community and hospital settings. The recommendations made by nurses, based on their assessments, have huge implications for clients. The study was carried out in south-east London. A semi-structured questionnaire was used to gather information on aspects of the assessment process, and a vignette was used to ascertain how nurses assess their clients for long-term care. The questionnaires were sent to 45 district nurses and elderly care nurses involved in assessing clients for long-term care. There was wide variation in nurses' assessments and opinions on the appropriate timing of the assessments. Nurses did not identify the importance of allowing individuals the opportunity to rehabilitate and recuperate before the nursing assessment and so made potentially inappropriate recommendations. The nurses highlighted that they were given little formal training on how to complete assessments and had limited knowledge about the different care options available for their clients. Nurses had differing opinions on who should carry out particular care tasks--registered nurse or carer. This has implications for the determination of whether a client is entitled to, and the level of, free nursing care. The variations in nurses' assessments indicated the importance of having more formal structures in place to train and support nurses undertaking this potentially life-changing work. PMID- 12574730 TI - Care management revisited: a follow-up study. AB - This article describes a follow-up to a study of community nurse care managers, undertaken in 1993-1994, in the light of the community care policy of the time. The original research found care management to be a viable practice model for community nurses to adopt, given the contemporary context; however, its long-term future was debatable. The present study comprised a questionnaire sent to two groups of community nurse care managers and their managers, who had participated in the original research. Responses indicated that a large proportion remained in a care management role some 4-5 years later, with the mental health and learning disability specialties being better represented than district nursing. Once again, survival was found to be dependent on a number of related contextual variables, which are discussed. The study concluded that the philosophy and principles of care management are still relevant to today's community nursing practice, even though the actual title appears to be less in evidence and current policy changes may render it less viable for the future. PMID- 12574731 TI - Is modified cow's milk formula effective in reducing symptoms of infant colic? AB - To determine the effectiveness of modified cow's milk formulas on the symptoms of infant colic, a mini-review (Griffiths, 2002) of double-blind randomized controlled trials was undertaken. The population was infants diagnosed with colic and the outcome was a reduction in infant distress. Medline, Embase, CINAHL and the Cochrane Library were searched and seven clinical trials and two systematic reviews were identified. After applying inclusion criteria, two studies were examined in the review. One study compared the effects of a modified formula and a standard cow's milk formula on bottle-fed infants. The other examined the effects of a low-allergen diet on breast-fed and bottle-fed infants. Results could only be retrieved from one study, which showed that hydrolysed formulas have a positive effect on reducing the symptoms of infant colic although sample sizes were small and the magnitude of benefit unclear. There is evidence to support advice to parents of bottle-fed infants with colic to consider changing to such formulas. PMID- 12574732 TI - Evaluation of a child development centre in a rural area. AB - In this article we discuss the setting up of a child development centre for children with a developmental delay in a rural market town away from a tertiary centre. The process was evaluated by a variety of methods including analysis of centre activity, preliminary questionnaires to parents at the end of their child's assessment, reflections by the centre team and in the final year of the project, a survey of all parents who had used the service, professionals who worked in it and other agencies in the centre town and a comparison town. Parents and staff reported high levels of satisfaction in relation to multidisciplinary working, venue, communication, and a child/family friendly approach. Further work is needed in developing links with social services and the voluntary sector. PMID- 12574733 TI - Older people as health service consumers: background to a study. AB - User involvement in health care has received a high profile within the NHS Plan (Department of Health, 2000a), giving patients a greater say in their treatment and in the way that the NHS works. Older people, i.e. those over 65 years of age, as a group, are the highest users of hospital and community services (DH, 2001a). The national service framework (NSF) for older people (DH, 2001a) highlights the need to listen to older people, help them make informed choices and involve them in achieving a healthy lifestyle. While the NSF for older people rightly attempts to ensure fair and equitable access to health care services for older people, it is assumed that they want to be involved, and want to exercise their rights as users. This assumption needs to be tested. This study, presented as a series of four articles, sets out to explore if there is a consumerist ethic among older people in relation to health care. The first paper outlines some of the literature reviewed together with the methodology. PMID- 12574734 TI - Closing service gaps will make a difference. PMID- 12574735 TI - Quantification of left ventricular volumes and ejection fraction using freehand transthoracic three-dimensional echocardiography: comparison with magnetic resonance imaging. AB - OBJECTIVES: Our aim was to validate 3-dimensional echocardiography (3DE) for assessment of left ventricular (LV) end-diastolic volume, end-systolic volume (ESV), stroke volume, and ejection fraction (EF) using the freehand-acquisition method. Furthermore, LV volumes by breath hold-versus free breathing-3DE acquisition were assessed and compared with magnetic resonance imaging (MRI). METHODS: From the apical position, a fan-like 3DE image was acquired during free breathing and another, thereafter, during breath hold. In 27 patients, 28 breath hold- and 24 free breathing-3DE images were acquired. A total of 17 patients underwent both MRI and 3DE. MRI contours were traced along the outer endocardial contour, including trabeculae, and along the inner endocardial contour, excluding trabeculae, from the LV volume. RESULTS: All 28 (100%) breath hold- and 86% of free breathing-3DE acquisitions could be analyzed. Intraobserver variation (percentual bias +/- 2 SD) of end-diastolic volume, ESV, stroke volume, and EF for breath-hold 3DE was, respectively, 0.3 +/- 10.2%, 0.3 +/- 14.6%, 0.1 +/- 18.4%, and -0.1 +/- 5.8%. For free-breathing 3DE, findings were similar. A significantly better interobserver variability, however, was observed for breath hold 3DE for ESV and EF. Comparison of breath-hold 3DE with MRI inner contour showed for end-diastolic volume, ESV, stroke volume, and EF, a percentual bias (+/- 2 SD) of, respectively, -13.5 +/- 26.9%, -17.7 +/- 47.8%, -10.6 +/- 43.6%, and -1.8 +/- 11.6%. Compared with the MRI outer contour, a significantly greater difference was observed, except for EF. CONCLUSIONS: 3DE using the freehand method is fast and highly reproducible for (serial) LV volume and EF measurement, and, hence, ideally suited for clinical decision making and trials. Breath-hold 3DE is superior to free-breathing 3DE regarding image quality and reproducibility. Compared with MRI, 3DE underestimates LV volumes, but not EF, which is mainly explained by differences in endocardial contour tracing by MRI (outer contour) and 3DE (inner contour) of the trabecularized endocardium. Underestimation is reduced when breath-hold 3DE is compared with inner contour analysis of the MRI dataset. PMID- 12574736 TI - Rapid freehand scanning three-dimensional echocardiography: accurate measurement of left ventricular volumes and ejection fraction compared with quantitative gated scintigraphy. AB - This study was performed to assess clinical feasibility of rapid freehand scanning 3-dimensional echocardiography (3DE) for measuring left ventricular (LV) end-diastolic and -systolic volumes and ejection fraction using quantitative gated myocardial perfusion single photon emission computed tomography as the reference standard. We performed transthoracic 2-dimensional echocardiography and magnetic freehand 3DE using a harmonic imaging system in 15 patients. Data sets (3DE) were collected by slowly tilting the probe (fan-like scanning) in the apical position. The 3DE data were recorded in 10 to 20 seconds, and the analysis was performed within 2 minutes after transferring the raw digital ultrasound data from the scanner. For LV end-diastolic and -systolic volume measurements, there was a high correlation and good agreement (LV end-diastolic volume, r = 0.94, P <.0001, standard error of the estimates = 21.6 mL, bias = 6.7 mL; LV end-systolic volume, r = 0.96, P <.0001, standard error of the estimates = 14.8 mL, bias = 3.9 mL) between gated single photon emission computed tomography and 3DE. There was an overall underestimation of volumes with greater limits of agreement by 2 dimensional echocardiography. For LV ejection fraction, regression and agreement analysis also demonstrated high precision and accuracy (y = 0.82x + 5.1, r = 0.93, P <.001, standard error of the estimates = 7.6%, bias = 4.0%) by 3DE compared with 2-dimensional echocardiography. Rapid 3DE using a magnetic-field system provides precise and accurate measurements of LV volumes and ejection fraction in human beings PMID- 12574737 TI - Quantification of regional myocardial perfusion using semiautomated translation free analysis of contrast-enhanced power modulation images. AB - Quantitative analysis of myocardial perfusion is currently based on manual tracing and frame-by-frame realignment of regions of interest. We developed a technique for semiautomated identification of myocardial regions from power modulation images as a potential tool for quantification of myocardial contrast enhancement. This approach was tested in 13 anesthetized pigs during continuous intravenous infusion of contrast at baseline, left anterior descending coronary artery occlusion, and reperfusion. Regional pixel intensity was calculated for each consecutive end-systolic frame after a high-energy ultrasound impulse, and fitted with an exponential function. Perfusion defects caused by occlusion of left anterior descending coronary artery were confirmed by a significant decrease in both postimpulse steady-state intensity and the initial rate of contrast replenishment (P <.05), which were reversed with reperfusion. Automated measurements of myocardial intensity correlated highly with conventional manual tracing (r = 0.90 to 0.97), and resulted in improved signal-to-noise ratios. This technique allows translation-free quantification of regional myocardial perfusion, without the need for manual tracing. PMID- 12574738 TI - Contrast-enhanced Doppler hemodynamics for noninvasive assessment of patients with chronic heart failure and left ventricular systolic dysfunction. AB - We sought to evaluate whether contrast-enhanced Doppler echocardiography can improve the noninvasive estimation of hemodynamic variables in left ventricular (LV) dysfunction. Right-heart catheterization and Doppler echocardiography were simultaneously performed in 45 patients with LV dysfunction (ejection fraction: 29 +/- 7%) in sinus rhythm. Noninvasive variables were estimated as follows: cardiac output by pulsed Doppler of LV outflow tract; pulmonary capillary wedge pressure by a regression equation including mitral and pulmonary venous flow variables; pulmonary artery mean pressure from the calculated systolic and diastolic pulmonary artery pressures; and pulmonary vascular resistance from the previous measurements according to hemodynamic definition. Contrast enhancement increased the feasibility of pulmonary capillary wedge pressure estimation from 60% to 100%; of pulmonary artery mean pressure from 42% to 91%; and of pulmonary vascular resistance from 42% to 91%. Strong correlations between invasive and noninvasive hemodynamic variables were found: r = 0.90, standard error of the estimate (SEE) 0.45 L/min for cardiac output; r = 0.90, SEE 3.1 mm Hg for pulmonary capillary wedge pressure; r = 0.93, SEE 3.7 mm Hg for pulmonary artery mean pressure; and r = 0.85 SEE 1.0 Wood units for pulmonary vascular resistance. Weaker correlations for PAMP (r = 0.82, SEE 5.6 mm Hg) and PVR (r = 0.66, SEE 1.7 Wood units) were apparent prior to contrast enhancement. When patients were separated according to PVR threshold values, the contrast allowed the correct placement of 88% of patients, whereas only 57% were correctly assigned without it. The contrast increased accuracy and reduced interobserver variability in the evaluation of hemodynamic variables. The contrast-enhanced study is capable of increasing the value of noninvasive hemodynamic assessment in LV dysfunction. PMID- 12574739 TI - Effects of dobutamine on left ventricular shape and geometry: an easy way to detect the functional status of chronic heart failure in patients with dilated cardiomyopathy. AB - OBJECTIVE: Changes in left ventricular geometry and shape are prominent characteristics of cardiomyopathy. The aim of this study was to investigate left ventricular geometry and shape changes after inotropic stimulation in patients with chronic nonischemic dilated cardiomyopathy. METHODS: A low infusion rate echocardiography-dobutamine study was performed in 35 consecutive patients (age: 50 +/- 8 years) with known dilated cardiomyopathy. Peak exercise oxygen consumption was measured the day before. Overall left ventricular chamber shape was assessed by using the long-/minor-axis dimension ratio obtained at end diastole and end-systole. RESULTS: After dobutamine infusion, in the entire study group an increase in systolic long-/minor-axis dimension ratio (0.11 +/- 0.13 mm, P <.001) was observed. The study population was further divided according to whether peak exercise oxygen consumption was less (group I) or more (group II) than 14 mL/kg/min. Although at baseline systolic long-/minor-axis dimension ratio was similar between groups, after dobutamine infusion it was higher in group II (1.51 +/- 0.19) than in group I (1.31 +/- 0.2, P <.01) indicating the abnormal response of systolic sphericity index to inotropic stimulation in patients with advanced functional class heart failure. CONCLUSION: The response of systolic sphericity index to dobutamine infusion might be an alternative index of the functional status of the failing heart in patients with nonischemic dilated cardiomyopathy. PMID- 12574740 TI - Moderate-dose dobutamine maximizes left ventricular contractile response during dobutamine stress echocardiography in children. AB - BACKGROUND: Assessment of ventricular contractile reserve by dobutamine stress echocardiography (DSE) may be a powerful tool for detection of subclinical ventricular dysfunction, however, the hemodynamic dose-response relationship during DSE in children has not been established. METHODS: To characterize changes in hemodynamics and ventricular contractility during DSE in children, 26 participants (age 8.3 +/- 4.8 years; 17 male/9 female) with normal resting left ventricular function underwent DSE. Participants with abnormal wall motion at rest or during DSE, or rejection were excluded. Left ventricular M-mode echocardiography and carotid pulse tracings were obtained at each stage for calculation of shortening fraction, velocity of circumferential fiber shortening (VCFc), and end-systolic wall stress (WS). Contractility was expressed as the difference between actual and predicted VCFc for measured WS. Dose-response curves for shortening fraction, VCFc, WS, and contractility (the difference between actual and predicted VCFc for measured WS) were obtained. RESULTS: Stepwise changes in contractility, systolic blood pressure, WS, and left ventricular shortening fraction were observed at doses up to, but not beyond, 20 microg/kg/min. Increases in double product were observed at doses up to 30 microg/kg/min. CONCLUSIONS: DSE at a dobutamine dose of 20 microg/kg/min is optimal to fully assess contractile reserve in children. Lesser doses may provide insufficient stress, whereas higher doses may incur unnecessary increases in myocardial oxygen consumption and side effects. PMID- 12574741 TI - Quantitative assessment of pericardial effusion volume by two-dimensional echocardiography. AB - The clinical applicability of a method for quantifying pericardial effusion (PE) was studied. The pericardial and cardiac volumes were determined from border tracings of 2-dimensional echocardiograms fitted with 3-dimensional disk models. The PE volume was the difference between pericardial and cardiac volumes. A phantom study included 54 cases with different volumes and viewing geometry showed a correlation coefficient r = 0.98 and an accuracy of +/- 6%. A clinical study included 20 cases showing that the estimated PE volume correlated well with that drained surgically (between 100 mL and 1200 mL): y = 0.81 x + 120 mL; r = 0.91, P <.0001. The percent error, determined by the standard error of the estimate (114 mL) over mean (548 mL), was 20%. Intraobserver variability was 4% and interobserver variability 6%. The 3-dimensional disk method provides a quick and convenient way to quantify PE from 2-dimensional echocardiograms with acceptable accuracy and reproducibility. PMID- 12574742 TI - Different involvement of right ventricular myocardial function in either physiologic or pathologic left ventricular hypertrophy: a Doppler tissue study. AB - The aim of the study was to analyze right ventricular (RV) myocardial function in patients with left ventricular (LV) hypertrophy secondary to either hypertrophic cardiomyopathy (HC) or athletic endurance training. Doppler echocardiography and pulsed Doppler tissue imaging of the posterior septal wall, and mitral and tricuspid annulus were performed in 32 top-level endurance athletes (AT) and in 27 patients with HC, all men. LV mass index was comparable between the 2 groups. All transmitral Doppler indexes were higher in AT, whereas only tricuspid inflow peak E and E/A ratio were slightly decreased in the HC group. In the HC group, Doppler tissue analysis showed lower myocardial systolic and early-diastolic (Em) peak velocities, and longer time intervals at the level of all the analyzed segments, even after correction for age, heart rate, and LV mass index. Distinct multiple linear regression models revealed an independent positive association between RV peak Em velocity and LV end-diastolic diameter (beta coefficient = 0.72, P <.0001) in AT, and an independent inverse correlation of the same peak Em velocity of tricuspid annulus with septal thickness (beta = - 0.65, P <.001) in the HC group. Of interest, a RV Em peak velocity < 0.16 m/s differentiated AT and HC groups better than tricuspid Doppler (89% sensitivity and 93% specificity). In conclusion, Em RV myocardial function is positively influenced by preload increase in AT and negatively associated to increased septal thickness in patients with HC. Therefore, Doppler tissue imaging may represent a useful tool in the differential diagnosis between athlete's heart and HC, underlining the different involvement of RV myocardial function in either physiologic or pathologic LV hypertrophy. PMID- 12574743 TI - Insights into the effect of aortic compliance on Doppler diastolic flow patterns seen in coarctation of the aorta: a numeric study. AB - BACKGROUND: In the echocardiographic evaluation of coarctation of the aorta, the degree of antegrade diastolic flow (diastolic runoff) noted on spectral Doppler tracings traditionally was thought to be solely dependent on lesion severity. However, recent in vitro experiments suggest the presence of this spectral Doppler pattern is as much related to the severity of coarctation as it is with changes in aortic compliance. Using state-of-the-art, multidisciplinary, numeric analysis tools, the purpose of this study was to investigate the specific fluid and wall mechanics present in coarctation of the aorta to further understand these relationships. METHODS: Three computational numeric models of coarctation were developed with high, low, and no wall compliance. Flow simulations were run representing high- and low-flow states. RESULTS: In both the low- and high-flow states, the degree of diastolic runoff increased with increasing vessel compliance. The high compliance model had larger changes in aortic dilatation in the precoarctation region compared with the low compliance model. CONCLUSIONS: Increased aortic compliance brings about greater dilatation of the precoarctation aorta in systole, resulting in a persistence of stored upstream energy. This stored energy, released downstream in diastole as the precoarctation aortic walls contract, leads to increased degrees of diastolic runoff. Numeric methods offer a unique perspective into the mechanisms behind such clinical measures. PMID- 12574744 TI - Benzocaine-induced methemoglobinemia: experience from a high-volume transesophageal echocardiography laboratory. AB - Benzocaine (ethyl aminobenzoate), a topical anesthetic widely used before transesophageal echocardiography, has been reported to cause acquired methemoglobinemia. The incidence of benzocaine-induced methemoglobinemia in clinical practice, however, has been difficult to estimate. After systematic review of our institutional experience for clinically recognized cases of benzocaine-induced methemoglobinemia in patients undergoing transesophageal echocardiography, we report an estimated incidence of 0.115% (95% confidence interval 0.037-0.269). This report also provides the largest analysis of the incidence of methemoglobinemia in readministration cases. Although controversy remains as to whether this is an idiosyncratic versus dose-related response, a description of purported patient risk factors is included. PMID- 12574745 TI - Diastolic dysfunction in an unusual case of cardiomyopathy in a child: insights from Doppler and Doppler tissue imaging analysis. AB - OBJECTIVES: This case report describes an unusual form of cardiomyopathy in a child with abnormalities in both ventricular relaxation and compliance. The diastolic abnormalities are well-defined using Doppler inflow velocities and Doppler tissue imaging, and the Doppler findings are correlated with intracardiac pressure measurements. This case illustrates the use of noninvasive Doppler techniques in identifying diastolic dysfunction, and it sheds additional light on the interplay of relaxation, compliance, and heart rate in creating the variable inflow Doppler patterns encountered clinically. PMID- 12574746 TI - Penetrating aortic atherosclerotic ulcer complicated by periesophageal hematoma. AB - Penetrating aortic atherosclerotic ulcers have been recently recognized as an entity among the acute aortic syndromes with a potentially fatal outcome. We describe the case of a patient presenting with severe chest pain who died as a result of a thoracic-aorta penetrating atherosclerotic ulcer complicated by a intramural hematoma of the esophagus and stomach, leading to exsanguination. To our knowledge this is the first case reported in the literature of such a complication from penetrating aortic atherosclerotic ulcers. PMID- 12574747 TI - Bilateral pulmonary artery compression and obstruction by tumor: diagnosis by unusual Doppler flow patterns. AB - Pulmonary artery obstruction may be caused by tumor within or external to the arteries. Presented here is a patient with life-threatening compromise in pulmonary flow that was caused by a pulmonary neoplasm. The Doppler echocardiogram showed subtotal narrowing of the right pulmonary artery and total occlusion of the left pulmonary artery. In addition, the beneficial effects of chemotherapy were documented by Doppler. PMID- 12574748 TI - An unusual cause of hemolysis in a patient with an aortic valved conduit replacement. AB - Hemolytic anemia is a well-known but uncommon complication in patients with prosthetic heart valves. It is most commonly a result of prosthetic valve dysfunction, periprosthetic valvular regurgitation, or both. We report a case of a 41-year-old man who had a previous aortic valve and root replacement for acute proximal aortic dissection, now presenting with hemolytic anemia. This was a result of flow obstruction at the distal anastomosis of the aortic conduit by the presence of multiple dissection flaps resulting in severe flow turbulence. Although the pathology was at the blind spot for transesophageal echocardiography, the dissection flaps, the flow turbulence, and the degree of obstruction were well-demonstrated by this technique after careful manipulation of the probe and a high index of suspicion. PMID- 12574749 TI - Mycotic aneurysm of the left ventricle: echocardiographic diagnosis. AB - We report an unusual case of myocardial mycotic aneurysm of the left ventricle resulting from a healed myocardial abscess caused by an aortic regurgitant jet lesion. The diagnosis was made during intraoperative transesophageal echocardiography and confirmed by surgical inspection. The echocardiographic features are described. PMID- 12574750 TI - A rare case of Alstrom syndrome presenting with rapidly progressive severe dilated cardiomyopathy diagnosed by echocardiography. AB - Alstrom Syndrome is an extremely rare, autosomal recessive genetic disorder characterized by a group of signs and symptoms including infantile onset dilated cardiomyopathy, blindness, hearing impairment/loss, and obesity; with diabetes, and hepatic and renal dysfunction later on in life. Since the first description of the syndrome in 1959, there have been fewer than 100 reported cases in the world. We report a case of a 7-month-old girl whose initial presentation of Alstrom syndrome was a rapidly progressive severe dilated cardiomyopathy, diagnosed by echocardiography. Unique characteristics of the cardiomyopathy in Alstrom syndrome and the important role of echocardiography in Alstrom syndrome and other genetically transmitted disorders, especially with known cardiovascular manifestations, is discussed. PMID- 12574751 TI - Uncertainty-based medicine. PMID- 12574752 TI - Image-guided sinus surgery: lessons learned from the first 1000 cases. AB - OBJECTIVE: To study physician utilization and experience with image-guidance technology for sinus surgery. STUDY DESIGN AND SETTING: Retrospective review of the first 1000 image-guided sinus operations performed by 42 surgeons at an academic medical center. RESULTS: Utilization of image-guidance systems showed a dramatic increase in both number of cases performed and surgeons who used this equipment (70.6% and 92.8%, respectively) during the first 2 years of its availability. Surgical volume subsequently decreased by a mean of 9.3% per year, whereas the number of surgeons using this technology plateaued. The majority of surgeons continued to perform image-guided surgery throughout the study period for selected cases. The knowledge base gained from this experience can best be summarized as a series of lessons learned. CONCLUSION: It is likely that the availability and utilization of image-guidance systems for sinus surgery will continue to increase in the future. Physicians who learn to use this new technology must do so with an appreciation for both its potential benefits and pitfalls. PMID- 12574753 TI - Allergy and chronic rhinosinusitis. AB - Allergy and its relationship to chronic rhinosinusitis are important contemporary topics in the practice of otolaryngology. The relationship may go unrecognized. It is important to understand the current knowledge. PMID- 12574754 TI - Quality of life for children with persistent sinonasal symptoms. AB - OBJECTIVE: The goal was to validate the SN-5 survey as a measure of longitudinal change in health-related quality of life (HRQoL) for children with persistent sinonasal symptoms. DESIGN AND SETTING: We conducted a before and after study of 85 children aged 2 to 12 years in a metropolitan pediatric otolaryngology practice. Caregivers completed the SN-5 survey at entry and at least 4 weeks later. The survey included 5 symptom-cluster items covering the domains of sinus infection, nasal obstruction, allergy symptoms, emotional distress, and activity limitations. RESULTS: Good test-retest reliability (R = 0.70) was obtained for the overall SN-5 score and the individual survey items (R >/= 0.58). The mean baseline SN-5 score was 3.8 (SD, 1.0) of a maximum of 7.0, with higher scores indicating poorer HRQoL. All SN-5 items had adequate correlation (R >/= 0.36) with external constructs. The mean change in SN-5 score after routine clinical care was 0.88 (SD, 1.19) with an effect size of 0.74 indicating good responsiveness to longitudinal change. The change scores correlated appropriately with changes in related external constructs (R >/= 0.42). CONCLUSIONS: The SN-5 is a valid, reliable, and responsive measure of HRQoL for children with persistent sinonasal symptoms, suitable for use in outcomes studies and routine clinical care. PMID- 12574755 TI - The role of rhinitis in chronic otitis media. AB - OBJECTIVE: We investigated the role of allergic rhinitis in chronic otitis media (otitis media with effusion [OME] and chronic perforation of the tympanic membrane) in Sao Paulo, Brazil and whether there is any association between these diseases. STUDY DESIGN AND SETTING: We studied 51 patients followed in the otologic group of the Otorhinolaryngology Division of the University of Sao Paulo Hospital. The patients were divided into 3 groups: allergic rhinitis, nonallergic rhinitis with eosinophils syndrome (NARES), and patients with types of rhinitis or without rhinitis. We analyzed the age, gender, intensity of the nasal disease, surgical procedure, and surgical results in each group. RESULTS: We found about 50% of patients presenting with nasal disease and nasal eosinophilia (33.33% of allergic and 15.69% of NARES). CONCLUSION: Nasal disease has an impact on otologic middle ear disease, considering that the normal nasal mucosa do not have eosinophils. PMID- 12574756 TI - Significance of empty sella in cerebrospinal fluid leaks. AB - OBJECTIVE: The role of elevated cerebrospinal fluid (CSF) pressures in the pathophysiology of various CSF leaks is not clear. Empty sella syndrome (ESS) is a radiographic finding that can be associated with elevated CSF pressures and may represent a radiographic indicator of intracranial hypertension. We present our experience with CSF leaks of various causes, the prevalence of ESS in the spontaneous and nonspontaneous categories, and the potential pathophysiology and unique management issues of the spontaneous CSF leak group. METHODS: We conducted a retrospective review of medical records, imaging studies, and surgical treatment of CSF leaks in patients treated by the senior author. RESULTS: Sixteen patients with spontaneous CSF leaks and 12 patients with nonspontaneous CSF leaks were surgically treated from 1996 through 2002. In the spontaneous group, 15 patients had complete imaging of the sella turcica. Ten had completely empty sellae and 5 had partially empty sellae, for a total of 100% (15 of 15). In the nonspontaneous group, 9 patients had complete imaging of the sella. Only 11% (1 of 9) had a partially empty sella and that was a congenital leak. Comparison of proportions between these 2 groups was significant (P = 0.01). The spontaneous group consisted primarily of obese, middle-aged females (13 of 16 patients). CONCLUSION: Empty sella probably represents a sign of elevated intracranial pressure that leads to idiopathic, spontaneous CSF leaks. Spontaneous CSF leaks are strongly associated with the radiographic finding of an empty sella and are more common in obese females, similar to benign intracranial hypertension. This unique population may require more aggressive surgical and medical treatment to prevent recurrent or multiple leaks. PMID- 12574757 TI - Management of osteomyelitis of the anterior skull base and craniovertebral junction. AB - OBJECTIVES: The study goals were to determine the patient demographics, identify predisposing factors, and determine efficacy of treatment for nonotologic osteomyelitis of the skull base and craniovertebral junction. STUDY DESIGN: All patients with a biopsy-proven diagnosis of osteomyelitis of the skull base treated by the author from 1997 through 2001 were retrospectively evaluated. RESULTS: Six patients were identified on review. The average age at presentation was 56.7 years (age range, 38 to 70 years), and all except one patient had an underlying immunocompromising condition (diabetes mellitus, human immunodeficiency virus infection, steroid use). Most presented with neurologic deficits associated with a destructive lesion of the osseous skull base. Aggressive debridement of involved bone enabled through the use of broad field standard skull base approaches was associated with clinical resolution of symptoms in each case. Systemic antibacterial/antifungal therapy and medical optimization remain important adjuncts in the treatment of this group of patients. CONCLUSION: The diagnosis of this entity may be difficult to discern from neoplastic involvement of the skull base. Standard skull base approaches are useful for both the diagnosis and the treatment of nonotologic osteomyelitis. PMID- 12574758 TI - The role of hair shaving in skull base surgery. AB - OBJECTIVE: We sought to evaluate surgical wound infection rates in patients undergoing skull base surgery without hair removal. METHODS: We undertook a retrospective study of 175 skull base operations performed without hair removal. Anterior operations were conducted via the subcranial approach (n = 120) and lateral or posterior procedures via various approaches (n = 55). Wounds were examined daily during hospitalization and at routine outpatient follow-up (8 to 45 months) and classified according to the Center for Disease Control and Prevention guidelines. RESULTS: The overall surgical wound infection rate was 1.1% (2 of 175): 0.8% (1 of 120) for anterior and 1.8% (1 of 55) for lateral or posterior procedures. It was similar for clean operations (lateral and posterior) and clean-contaminated (anterior) procedures and was less than or similar to the rates reported for skull base procedures with hair removal. No wound infection occurred among the infected (trauma, fungal infections, and brain abscess) patients. CONCLUSIONS: Skull base surgery without hair removal is safe and not associated with increased risk of wound infection. The method may prevent additional psychologic stress, promote restoration of the patient's self-image, and accelerate his or her return to normal life. PMID- 12574759 TI - Web-based clinical decision support system for triage of vestibular patients. AB - OBJECTIVES: We sought to use a clinical decision support system (CDSS) to save costs and to improve scheduling of vestibular patients in an otolaryngology clinic. STUDY DESIGN AND SETTING: We conducted a concurrent review of 50 vestibular patients scheduled in the University of Missouri otolaryngology clinic with or without testing based on the outcome of a CDSS. The CDSS was implemented using Web-based technology. Charges incurred by the health care system through tests determined by the CDSS were compared with those incurred using the standard procedure of ordering hearing tests and electronystagmography for all patients. RESULTS: Thirty-nine tests were prescheduled using the CDSS. Twenty-five additional tests were ordered after the visit. The CDSS resulted in savings of $37,904.00 in charges to the health care system. The CDSS showed high specificity and variable sensitivity. CONCLUSION: A Web-based CDSS can be used to better manage and coordinate patient encounters. SIGNIFICANCE: One important reason to use a CDSS in health care management is to lower costs. PMID- 12574760 TI - A practical assessment algorithm for diagnosis of dizziness. AB - OBJECTIVE: We sought to test a 3-parameter model for diagnosis of dizziness based on the type and temporal characteristics of the dizziness and on hearing status. STUDY DESIGN AND SETTING: We conducted a prospective blinded study at a tertiary referral neurotology practice. Before examination, patients completed a questionnaire reporting type and timing of dizziness symptoms and hearing status. Clinical diagnoses were compared with questionnaire results. RESULTS: Fifty-seven patients completed the questionnaire. We were able to correctly classify 21 (60%) of the 35 subjects who had a common otogenic cause of vertigo by the diagnostic algorithm. CONCLUSION: A simple classification of dizziness by type, timing, and hearing status can be self-reported by patients using a brief questionnaire. This classification scheme is as good as others of much greater complexity. SIGNIFICANCE: The simple classification scheme reported here is based on history alone and facilitates triage of dizzy patients into diagnostic groups for work-up and management. PMID- 12574761 TI - Increased independence and decreased vertigo after vestibular rehabilitation. AB - OBJECTIVE: We sought to determine the effectiveness in decreasing some symptoms, such as vertigo, and increasing performance of daily life skills after vestibular rehabilitation. STUDY DESIGN AND SETTING: Patients who had chronic vertigo due to peripheral vestibular impairments were seen at a tertiary care center. They were referred for vestibular rehabilitation and were assessed on vertigo intensity and frequency with the use of the Vertigo Symptom Scale, the Vertigo Handicap Questionnaire, the Vestibular Disorders Activities of Daily Living Scale, and the Dizziness Handicap Inventory. They were then randomly assigned to 1 of 3 home program treatment groups. RESULTS: Vertigo decreased and independence in activities of daily living improved significantly. Improvement was not affected by age, gender, or history of vertigo. CONCLUSION: For many patients a simple home program of vestibular habituation head movement exercises is related to reduction in symptoms and increasing independence in activities of daily living. PMID- 12574762 TI - Task performance in stapedotomy: comparison between surgeons of different experience levels. AB - OBJECTIVES: Two steps in stapedotomy are particularly challenging: (1) micropick fenestration of the stapes footplate (SF) and (2) crimping of the stapes prosthesis (SP) to the incus. We conducted trials to determine if experience correlates with differences in performance for these tasks. METHODS: In a surgical model of stapedotomy, performance was measured for 3 experienced and 3 novice surgeons. For fenestration, we measured ability to target the fenestration and force applied to the SF. For crimping, we measured crimp quality, movement of the SP during crimping, and force applied to the SF. RESULTS: Experienced surgeons demonstrated significantly better ability to target the fenestration and, during crimping, caused less SP movement and a significantly lower rate of SP dislodgment. CONCLUSIONS: Clear differences in task performance are measurable between more and less experienced surgeons during critical steps of stapedotomy. CLINICAL SIGNIFICANCE: The observed differences in task performance may contribute to an understanding of maneuvers that increase the risk of inadequate prosthesis placement and cochlear trauma-factors likely responsible for variable hearing results with strapedotomy. PMID- 12574763 TI - Establishment of normative cochlear and vestibular measurements to aid in the diagnosis of inner ear malformations. AB - OBJECTIVE: We sought to establish normative measurements of the inner ear using computed tomography (CT) of the temporal bone to aid in the diagnosis of inner ear malformations. STUDY DESIGN AND SETTING: Prospective measurements of the inner ear structures were made on axial and coronal temporal bone CT scans on 15 patients with normal hearing and 15 patients with sensorineural hearing loss. RESULTS: The vertical height of the cochlea on coronal scan and the size of the central bony island within the lateral semicircular canal on axial scan along with visual inspection identified 7 inner ear abnormalities in 6 patients: 5 cases of lateral semicircular canal dysplasia and 2 cases of cochlear hypoplasia. In contrast, visual inspection alone identified only 4 of the 7 abnormalities. CONCLUSIONS AND SIGNIFICANCE: Routine measurement of the cochlear height and bony island of the lateral semicircular canal, in conjunction with visual inspection of CT images, will increase recognition of common inner ear malformations. PMID- 12574764 TI - Autologous fat grafting for the refractory patulous eustachian tube. AB - The patulous eustachian tube (pET) presents a challenging management problem. Patients with the pET disorder are disturbed to the point of mental illness by their symptoms of aural fullness and autophony, especially the echo of their own voice. Thus far, described treatment methods are often temporary or ineffective. Here, we describe a minimally invasive method for the treatment of pET that involves cauterization with autologous fat graft plugging of the ET at its nasopharyngeal orifice, in conjunction with myringotomy and ventilation tube placement. This procedure has been successful for the treatment of refractory pET in 2 patients at the House Ear Clinic with at least 1-year follow-up and we propose that it may accomplish relief of symptoms by permanent stenosis of the ET. PMID- 12574765 TI - Diagnosis and treatment of sudden-onset sensorineural hearing loss: a study of 51 patients. AB - OBJECTIVE: The study purpose was to determine the efficacy of steroid and antiviral therapy in the management of idiopathic sudden sensorineural hearing loss (SSNHL). STUDY DESIGN AND SETTING: We conducted a retrospective study of patients presenting to an academic tertiary care center. Fifty-one patients were evaluated. All patients were placed on the same treatment protocol. RESULTS: Thirty-seven patients (73%) had recovery of hearing. Ninety-one percent of patients with vertigo and all patients with mid-frequency hearing loss and up sloping hearing loss recovered with treatment (P < 0.05). Recovery was significantly related to age, onset of hearing loss, and audiogram type; however outcome was not significantly related to gender, vertigo, tinnitus, or laterality (P < 0.05). CONCLUSION: Our treatment protocol produced a recovery rate, which exceeds the spontaneous recovery rate. Unlike prior studies, all patients with up sloping and mid-frequency SSNHL had recovery. In addition, vertigo did not indicate a poor prognosis. SIGNIFICANCE: Antiviral therapy and increased length of steroid treatment may play a role in the improved recovery rates. PMID- 12574766 TI - The association between laryngopharyngeal sensory deficits, pharyngeal motor function, and the prevalence of aspiration with thin liquids. AB - OBJECTIVE: The study goal was to evaluate the association among laryngopharyngeal sensory deficits, pharyngeal motor function, and the prevalence of aspiration with thin liquids. STUDY DESIGN AND SETTING: We conducted a prospective study of 204 consecutive patients undergoing flexible endoscopic evaluation of swallowing with sensory testing and an assessment of pharyngeal motor function (pharyngeal squeeze). Patients were divided into 6 groups depending on the results of sensory and motor testing in the laryngopharynx. Subjects were given 5 mL of thin liquid, and the prevalence of aspiration in each group was compared. RESULTS: The mean age of the entire cohort was 65 years (58% female). The prevalence of aspiration in patients with intact laryngopharyngeal sensation was 2% (3 of 137) in persons with intact pharyngeal motor function and 29% (2 of 7) when pharyngeal motor function was impaired (P < 0.05). The prevalence of aspiration in patients with a moderate decrease in laryngopharyngeal sensation was 0% (0 of 9) in persons with intact pharyngeal motor function and 67% (2 of 3) when pharyngeal motor function was impaired (P < 0.05). The prevalence of aspiration in patients with severely diminished or absent laryngopharyngeal sensation was 15% (5 of 33) in persons with intact pharyngeal motor function and 100% (15 of 15) when pharyngeal motor function was impaired (P < 0.05). CONCLUSION: Patients with severely diminished laryngopharyngeal sensation and pharyngeal motor function are at an extremely high risk of aspirating thin liquids (100%). Moderate sensory deficits only appear to influence the prevalence of thin liquid aspiration in the presence of pharyngeal motor dysfunction. Severe laryngopharyngeal sensory deficits are associated with the aspiration of thin liquids regardless of the integrity of pharyngeal motor function. We assume that all persons with an insensate laryngopharynx aspirate thin liquids until proved otherwise. These results emphasize the relationship between laryngopharyngeal sensation and pharyngeal motor function in the evaluation of patients for suspected aspiration. PMID- 12574767 TI - Diagnosis of laryngopharyngeal reflux disease with digital imaging. AB - OBJECTIVE: The study goal was to assess the use of digital laryngeal videostroboscopy (LVS) in the detection of objective improvement of the larynx after 6 weeks of proton pump inhibitor (PPI) therapy and to evaluate a clinical grading scale for findings of laryngopharyngeal reflux disease (LPRD). STUDY DESIGN AND SETTING: We conducted a prospective analysis of digital LVS examinations from a tertiary referral center for laryngology by 3 independent blinded examiners. RESULTS: The mean score on the Laryngopharyngeal Reflux Disease Index for experimental patients was significantly higher than that for control patients (9.50 versus 2.92, P < 0.001), and posttreatment scores were significantly lower than pretreatment scores (7.35 versus 9.50, P < 0.001). Useful items on the grading scale for assessing the presence of LPRD and improvement in LPRD included supraglottic edema and erythema, glottic edema and erythema, and subglottic edema and erythema. Nonuseful items included the presence of pachydermia, granulomas, nodules, leukoplakia, and polyps. CONCLUSION: Objective improvement of signs of LPRD can be detected after 6 weeks of PPI therapy using digital LVS. The Laryngopharyngeal Reflux Disease Index is a useful valid clinical tool for following treatment response to PPI therapy. PMID- 12574768 TI - An analysis of time and staff utilization for open versus percutaneous tracheostomies. AB - OBJECTIVE: We examined staff utilization and procedure length for percutaneous and open bedside tracheostomies in an intensive care setting. STUDY DESIGN: Prospective clinical outcomes study. METHODS: Intensive care unit (ICU) tracheostomy consults meeting criteria for bedside procedures were randomized to open or percutaneous procedures. The Cook percutaneous kit and a prepackaged tracheostomy tray were used. ICU nursing and respiratory therapy staff was present for all procedures. The total resident time, staff time, and procedure length were recorded. Twelve patients underwent percutaneous tracheostomy, and 12 received an open tracheostomy. RESULTS: An operating room nurse was present for 7 of the open procedures. Ancillary medical staff was present for 3 open tracheostomies: anesthesia for 2 and critical care for 1. Ancillary medical staff was present for 4 percutaneous tracheostomies: anesthesia staff for 1 and critical care for 3. The average resident presence, staff presence, and procedure length for open tracheostomies were 47, 30, and 12 minutes, respectively. For percutaneous tracheostomies, the times were 39, 29, and 12 minutes, respectively. One intraoperative complication occurred during a percutaneous procedure and 2 perioperative complications occurred: 1 in the open group and 1 in the percutaneous group. CONCLUSIONS: There was no significant difference in procedure length, resident time, or staff time between the 2 procedures. Ancillary staff was occasionally used but was not thought to be necessary for the majority of procedures. Both procedures can be safely and expediently performed in the ICU. PMID- 12574769 TI - A population-based analysis of survival factors in differentiated and medullary thyroid carcinoma. AB - OBJECTIVE: The study purpose was to determine survival and prognostic factors for differentiated thyroid carcinoma (DTC). METHODS: Cases of DTC were extracted from the Surveillance, Epidemiology and End Results database from 1988 through 1998. Kaplan-Meier survival analysis was conducted for papillary, follicular, and medullary histologies. Cox proportional hazard analysis was used to examine the influence of age, gender, tumor size, local extension, and cervical node involvement on overall survival. RESULTS: A total of 18,118 cases were identified, including 15,820 (87.3%) papillary carcinomas, 1799 (9.9%) follicular carcinomas, and 499 (2.8%) medullary carcinomas. Mean survival (10-year survival) was 122 (87.7%), 117 (80.2%), and 108 (73.7%) months for papillary, follicular, and medullary tumors, respectively. For each histology, increasing age, male gender, and degree of local extension substantially reduced survival. Cervical metastasis did not influence survival for papillary or follicular carcinomas but approached significance for medullary carcinoma (P = 0.065). CONCLUSIONS: Degree of local extension in thyroid carcinoma should be subclassified to more accurately determine prognosis. Treatment of the neck should be considered for medullary thyroid carcinoma. PMID- 12574770 TI - Development of interface in hydroxyapatite implanted in the middle ear of the rat: a light and scanning microscopy study. AB - OBJECTIVE: The aim of this work is to study the chronology in the biological covering of the hydroxyapatite bioprostheses in the healthy middle ear of the rat. STUDY DESIGN AND SETTING: Dense hydroxyapatite is implanted into the middle ear of 45 Long-Evans rats, between stapes and tympanic membrane. The sample was divided into 3 groups for morphologic, functional, and chemical studies at 15 days, 3 months, and 6 months after surgery, respectively. RESULTS: The short-term studies reveal that the prostheses is surrounded by a fibrous epithelial tissue referred as the interface; it presents a marked inflammatory reaction that decreases gradually in the medium-term and long-term studies; in this way, we observe a progressive adhesion between implant and the contact tissues. CONCLUSIONS AND SIGNIFICANCE: This model has enabled us to study the cell reaction produced by the contact of the prostheses with bone and soft tissue and their impact on the functional qualities of the prostheses. PMID- 12574771 TI - Activity of matrix metalloproteinases-2 and -9 in advanced laryngeal cancer. AB - OBJECTIVES: Matrix metalloproteinases-2 and -9 (MMP-2 and MMP-9) are proteolytic enzymes that digest collagen type IV and other components of the basement membrane. They play a key role in local invasion and the formation of distant metastases by malignant tumors. The aim of this study was to evaluate the activity of MMP-2 and MMP-9 in stage III and IV laryngeal cancers. STUDY DESIGN: In the study we used specimens of laryngeal cancer and surrounding normal mucosa obtained from 23 patients undergoing surgical treatment as a primary therapy. After extraction of MMP-2 and MMP-9 from tissue samples, their activity was assessed with zymography. RESULTS: Greater activity of MMP-2 and MMP-9 and a higher active/latent MMP-2 ratio were found in cancer compared with normal mucosa. Moreover, N2 tumors revealed greater activity of MMP-2 in comparison with N1 and N0 tumors. CONCLUSIONS: Results of the study indicate that both MMP-2 and MMP-9 may be involved in the expansion of laryngeal cancer. MMP-2 may also play an important role in the lymphatic spread of some laryngeal tumors. PMID- 12574772 TI - Complications of mandibulotomy: midline versus paramidline. AB - OBJECTIVE: Mandibulotomy is an important surgical approach to oral cavity and oropharynx. The objective of this study was to evaluate the related complications of 2 common mandibulotomies: midline (osteotomy between 2 central incisors) and paramidline (osteotomy between lateral incisor and canine). STUDY DESIGN: Ninety six patients who had cancer in the oral cavity or oropharynx had preoperative evaluation of the mandible with panoramic films. Among them, 42 patients underwent mandibulotomies: midline for 19 patients and paramidline for 23. RESULTS: Twenty (47.6%) of 42 patients had mandibulotomy-related complications, including 9 (21.4%) minor complications and 11 (26.2%) major complications. There were 6 (31.6%) major and 4 (21.1%) minor complications in the midline mandibulotomy and both 5 (21.7%) major and minor complications in the paramidline mandibulotomy. CONCLUSION: There was no significant difference in the mandibulotomy-related complication rate between midline and paramidline mandibulotomies. Paramidline mandibulotomy, which preserves the geniohyoid and genioglossus muscles, should be a better function-preserving operation than midline mandibulotomy. PMID- 12574773 TI - Sequential upper airway changes during mandibular distraction for obstructive sleep apnea. PMID- 12574774 TI - Inflammatory myofibroblastic tumor in the neonate: a case report. PMID- 12574775 TI - Alveolar soft part sarcoma of the mandible. PMID- 12574776 TI - Massive facial angiofibroma in a patient with tuberous sclerosis. PMID- 12574777 TI - Occult foreign body aspiration in adults. PMID- 12574778 TI - Cervical subcutaneous emphysema after lower abdominal laparoscopic surgery. PMID- 12574779 TI - Salivary gland choristoma of the middle ear in a child: a case report. PMID- 12574780 TI - Concurrent intracranial and extracranial complications secondary to cholesteatoma: a case report. PMID- 12574781 TI - An unusual otomicroscopy diagnosis. PMID- 12574782 TI - Image-guided percutaneous biopsy of the adrenal gland: review of indications, technique, and complications. PMID- 12574783 TI - A pictorial review of conventional and new applications of duplex Doppler sonography in the pediatric abdomen. PMID- 12574784 TI - Better blood pressure control: how to combine drugs. AB - Prospective comparisons of different drug classes have shown that differences in blood pressure control, rather than differences between drug classes, have the over-riding influence on overall outcome. The same studies have also reinforced the need, in the majority of patients, to use combinations of drugs in order to achieve the target of <140/85 mmHg. By contrast, most patients in routine practice receive single agents and consequently fail to achieve target blood pressure. This failure reflects in part the emphasis in individual studies and subsequent guidelines on comparison of individual drugs. In this article we show how the consistency of both theory and a broad range of evidence permits a didactic approach to combination therapy. Our advice is based on the growing recognition that essential hypertension and its treatment fall into two main categories. Younger Caucasians usually have renin-dependent hypertension that responds well to angiotensin-converting-enzyme inhibition or angiotensin receptor blockade (A) or ss blockade (B). Most other patients have low-renin hypertension that responds better to calcium channel blockade (C) or diuretics (D). These latter drugs activate the renin system rendering patients responsive to the addition of renin suppressive therapy. Coincidence of the initials of these main drug classes with the first four letters of the alphabet permits an AB/CD rule, according to which recommended combinations are one drug from each of the "AB" and "CD" categories of drugs. However, the diabetogenic potential of the older "B" and "D" classes leads us to advise against combining "B" and "D" in older patients, and to recommend "A" + "C" + "D" as standard triple therapy for resistant hypertension. PMID- 12574785 TI - Normal blood pressure values as perceived by normotensive and hypertensive subjects. AB - In spite of considerable progress, the control of hypertension in most countries is still insufficient. One of the reasons may be the poor awareness of the blood pressure (BP) cutoff values that define hypertension. The aim of this report is to verify the hypothesis that patients with hypertension accept higher BP levels as normal. The study was performed during a street-based BP screening project carried out across Poland, in summer 1997. In 444 persons who voluntarily participated in Cracow's part of the project, BP was taken on the left arm, in the sitting position, after a minimum of 5 min of rest, using a semiautomated device (Digital Blood Pressure UA-702). A questionnaire investigated participants' age, weight, height, level of education, history of hypertension and perception of normal values of BP. We compared measured BP values with those perceived as normal, and with the values recommended by WHO/ISH guidelines (<140/90 mmHg). To analyse the data we used Student's t-test and linear regression with adjustment for age and body mass index (BMI). Hypertensive subjects, compared with normotensives, were less aware of normal BP values (47.4 vs 83.9%, P<0.001, for systotic blood pressure, and 77.4 vs 88.4%, P<0.01, for diastotic blood pressure). Measured BP was positively related to BP values stated as normal. A similar relation was observed for age and BMI. In conclusion, poor awareness of normal BP values in hypertensives can be an important factor hindering better BP control. Education strategies might prove to be highly effective in helping to tackle the epidemics of hypertension. PMID- 12574786 TI - Isolated systolic hypertension: data on a cohort of young subjects from a French working population (IHPAF). AB - Elderly patients with isolated systolic hypertension (ISH)--systolic blood pressure (SBP) > or =140 mmHg and diastolic blood pressure (DBP) <90 mmHg--have increased mortality and morbidity. The aim was to study the incidence of ISH in a younger population of between 15 and 60 years of age, and to measure pulse pressure (PP), mean arterial pressure (MAP) and heart rate (HR) in these subjects. The study population consisted of 27 783 subjects, aged 15-60 years, untreated for hypertension (HT) from a cohort of employees formed to study the incidence of HT in the French working population (AIHFP). BP and HR were measured with a validated, automatic device after 5, 6 and 7 min at rest. The prevalence of ISH was 6.9% in men, 2.3% in women. This prevalence was over 5% in young men and increased at 40-44 years; it was negligible in young women, but increased at 50-54 years to about 10% (ie to the same level as in men of the same age): PP in subjects with ISH (46.9 mmHg) was significantly higher than in the normotensive group (NT-40.9 mmHg); it was comparable in both young men (65.5 mmHg) and older men (66 mmHg); it was higher in men (63.1 mmHg) than in women (61.5 mmHg). HR was higher in ISH than in NT and it was higher in women ( approximately 5 bpm) in whom it decreased with age. The prevalence of ISH is not negligible in HT (30% men, 25% women), with a high prevalence in young subjects and elevated PP, MAP and HR values. These data should be taken into account as elevated ISH, PP and HR are considered as cardio-vascular risk factors. PMID- 12574787 TI - Change in cardiovascular risk profile by echocardiography in medium-risk elderly hypertensives. AB - It has been clearly demonstrated that ageing and arterial hypertension are both associated with an increased prevalence of left ventricular hypertrophy (LVH), which is a powerful risk factor for cardiovascular (CV) events. The objective of this study was to assess the impact of echocardiographic LVH in profiling the absolute CV risk stratification according to the 1999 World Health Organization International Society of Hypertension (WHO/ISH) guidelines in elderly hypertensive patients. A total of 223 never-treated elderly patients (> or =65 years) with essential hypertension (98 men, 125 women, mean age 72+/-5 years) referred to our outpatient hospital clinic were included in the study. They underwent the following procedures: (1) medical history, physical examination, and clinic blood pressure; (2) routine blood chemistry and urine analysis and (3) electrocardiogram. The risk was initially stratified according to the routine procedures suggested by WHO/ISH guidelines and subsequently reassessed by adding the results of echocardiography (LVH as left ventricular mass index >51 g/m(2.7) in men and >47g/m(2.7) in women). According to routine classification, 56% (n=125) were medium-risk patients, 29% (64) high-risk and 15% (34) very-high-risk patients. The overall prevalence of LVH was 56% (48% in medium-risk and 62% in high-risk or very-high-risk patients, P<0.01). A marked change in risk stratification was observed when echocardiographic LVH was taken into consideration: medium-risk patients decreased to 29% and high-risk patients rose to 56% (P<0.01). In conclusion, ultrasound assessment of cardiac target organ damage is extremely useful in obtaining a more valid assessment of global cardiovascular risk in elderly hypertensives, because stratification based on diagnostic routine procedures can underestimate the overall risk in a large fraction (48%) of medium-risk subjects. PMID- 12574788 TI - Bone mass and bone modelling markers in hypertensive postmenopausal women. AB - Numerous phosphocalcium alterations associated with bone mineral density in hypertension have been described, but very few studies assess them. This study assesses bone mass in hypertensive postmenopausal women and the hypertension influence determining both calcium homeostasis and bone turnover markers. Blood and urine samples were analysed for calcium metabolism-related parameters. Densitometry studies were conducted in the lumbar spine (L2-L4). Hypertensive osteoporotic women--selected from 82 women, with 22% osteoporosis prevalence, similar to the rate for the same age in the Spanish population--had significantly higher levels of body mass index (29+/-4 vs 26+/-4, P=0.019), calciuria (293+/ 146 vs 210+/-116 mg/24 h, P=0.023) and calcium/creatinine ratio (0.33+/-0.2 vs 0.22+/-0.1 P=0.003) vs hypertensive nonosteoporotic women. No relation was found between systolic and diastolic blood pressure with bone mass. However, there was a negative osteocalcin correlation (r=-0.386, P=0.0001, and r=-0.242, P=0.033). Calciuria is associated with bone mass decrease in hypertensive women, and there is no relation between bone mass and blood pressure. PMID- 12574789 TI - Assessment of cardiovascular reactivity by fractal and recurrence quantification analysis of heart rate and pulse transit time. AB - Methods used for the assessment of cardiovascular reactivity are flawed by nonlinear dynamics of the cardiovascular responses to stimuli. In an attempt to address this issue, we utilized a short postural challenge, recorded beat-to-beat heart rate (HR) and pulse transit time (PTT), assessed the data by fractal and recurrence quantification analysis, and processed the obtained variables by multivariate statistics. A 10-min supine phase of the head-up tilt test was followed by recording 600 cardiac cycles on tilt, that is, 5-10 min. Three groups of patients were studied, each including 20 subjects matched for age and gender- healthy subjects, patients with essential hypertension (HT), and patients with chronic fatigue syndrome (CFS). The latter group was studied on account of the well-known dysautonomia of CFS patients, which served as contrast against the cardiovascular reactivity of the healthy population. A total of 52 variables of the HR and PTT were determined in each subject. The multivariate model identified the best predictors for the assessment of reactivity of healthy subjects vs CFS. Based on these predictors, the "Fractal & Recurrence Analysis-based Score" (FRAS) was calculated: FRAS=76.2+0.04*HR-supine-DET -12.9*HR-tilt-R/L -0.31*HR-tilt-s.d. -19.27*PTT-tilt-R/L -9.42*PTT-tilt-WAVE. The median values and IQR of FRAS in the groups were: healthy=-1.85 (IQR 1.89), hypertensives=+0.52 (IQR 5.78), and CFS= 24.2 (5.34) (HT vs healthy subjects: P=0.0036; HT vs CFS: P<0.0001). Since the FRAS differed significantly between the three groups, it appears likely that the FRAS may recognize phenotypes of cardiovascular reactivity. PMID- 12574790 TI - Plasma levels of active extracellular matrix metalloproteinases 2 and 9 in patients with essential hypertension before and after antihypertensive treatment. AB - This study was designed to test the hypothesis that plasma concentrations of matrix metallo-proteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9), two enzymes that share similar substrate specificity (collagen type IV and V), possibly related to vascular remodelling, are altered in essential hypertension. The second aim of the study was to assess whether chronic antihypertensive treatment with the calcium channel blocker amlodipine would normalize these alterations. To test this hypothesis, we measured plasma concentrations of active MMP-2 and MMP-9 in 42 patients with never-treated essential hypertension and in 25 normotensive control subjects. Measurements were repeated after 6 months of treatment with the calcium channel blocker amlodipine. Baseline values of MMP-2 and MMP-9 were decreased (P=0.01 and 0.002, respectively) in hypertensive patients compared with normotensives. Hypertensive patients with systemic vascular resistances <1440 dyn s/cm(5) exhibited higher values of MMP-2 (P=0.005) and MMP-9 (P=0.001) than hypertensive patients with systemic vascular resistances >1440 dyn s/cm(5). Treated patients attained a nonsignificant increase in MMP-2 plasma concentrations, but a significant increase in MMP-9 plasma concentrations (P=0.01) compared to respective values before treatment. In conclusion, these findings suggest that plasma concentrations of active MMP-2 and MMP-9, mainly related to vascular extracellular matrix metabolism, are depressed in patients with essential hypertension. A 6 month treatment with amlodipine can normalize MMP-9 but not MMP-2 plasma concentrations. The hypothesis that antihypertensive treatment may modulate collagen metabolism remains to be determined by further studies. PMID- 12574791 TI - Subjects with essential hypertension are more sensitive to the inhibition of 11 beta-HSD by liquorice. AB - In this intervention study, we have investigated if hypertensive patients are more sensitive to liquorice-induced inhibition of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) type 2 than normotensive (NT) subjects and if the response depends on gender. Healthy volunteers and patients with essential hypertension (HT), consumed 100 g of liquorice daily, for 4 weeks, corresponding to a daily intake of 150 mg glycyrrhetinic acid. Office, 24-h ambulatory blood pressure (BP) and blood samples were measured before, during and after liquorice consumption. Effect on cortisol metabolism was evaluated by determining the urinary total cortisol metabolites and urinary free cortisol/free cortisone quotient (Q). The mean rise in systolic BP with office measurements after 4 weeks of liquorice consumption was 3.5 mmHg (p<0.06) in NT and 15.3 mmHg (p=0.003) in hypertensive subjects, the response being different (p=0.004). The mean rise in diastolic BP was 3.6 mmHg (p=0.01) in NT and 9.3 mmHg (p<0.001) in hypertensive subjects, the response also being different (p=0.03). Liquorice induced more pronounced clinical symptoms in women than in men (p=0.0008), although the difference in the effect on the BP was not significant. The increase in Q was prominent (p<0.0001) and correlated to the rise in BP (p=0.02). The rise in BP was not dependant on age, the change in plasma renin activity or weight. We conclude that patients with essential HT are more sensitive to the inhibition of 11 beta-HSD by liquorice than NT subjects, and that this inhibition causes more clinical symptoms in women than in men. PMID- 12574792 TI - Endothelin-1 and vasopressin plasma levels are not associated with the insertion/deletion polymorphism of the human angiotensin I-converting enzyme gene in patients with coronary artery disease. AB - The objective was to investigate whether the renin-angiotensin (RA) system and related peptides endothelin-1 (ET-1) and vasopressin (VP) influence the development of coronary artery disease (CAD). Angiotensin I-converting enzyme (ACE) insertion/deletion (I/D) gene polymorphism has been associated with the risk of CAD. The ACE I/D polymorphism determines ACE activity, but plasma levels of other RA system components remain unchanged. However, ET-1 and VP production could be increased by RA system-dependent stimulation, continually promoted by paracrine stimulation and sustained by neointimal growth. ET-1 and VP have not been associated with the ACE I/D polymorphism so far. The present study investigated the association of the ACE I/D polymorphism with plasma concentrations of ET-1 and VP, as well as with renin, angiotensin-II (AT-II) and ACE activity in 98 Caucasian individuals with CAD. ACE I/D polymorphism showed no association with plasma levels of VP, ET-1, AT-II or renin. These parameters were also not associated taking into consideration different patient variables, such as diabetes mellitus, hypertension or severity of CAD. Only plasma ACE activity was associated with the D allele. In conclusion, the ACE I/D polymorphism could not be related to plasma concentrations of VP, ET-1, renin or AT-II, but as previously demonstrated, it could only be related to ACE activity in patients with CAD. Differences in ACE activity between ACE I/D genotype subgroups are probably compensated within the RA system itself or within non-ACE pathways, so that plasma concentrations of the related peptides ET-1 and VP remain unaffected. PMID- 12574793 TI - Evaluation of the efficacy and tolerability of the combination delapril plus indapamide in the treatment of mild to moderate essential hypertension: a randomised, multicentre, controlled study. AB - The aim of the study was to evaluate efficacy and tolerability of two different fixed combinations of an angiotensin-converting enzyme inhibitor and a diuretic: delapril+indapamide (D+I) and captopril+hydrochlorothiazide (C+H) administered for 6 months to patients with mild to moderate essential hypertension. In all, 96 centres participated in this randomised, parallel groups, controlled study. A total of 829 patients with uncomplicated mild to moderate hypertension were randomised, and 790 were eligible for the analysis of efficacy (intention to treat). Patients of both sexes, aged 18-75 years, newly diagnosed or untreated during the last month were included in the study if their diastolic blood pressure (DBP) was > or =95 and < or =114 mmHg. The starting doses of the drugs were delapril 30 mg+indapamide 1.25 mg tablets o.d. or captopril 50 mg+hydrolchlorothiazide 15 mg tablets o.d. After a 1-month treatment period, nonresponders (DBP >90 mmHg, or decrease in DBP <10 mmHg) had the daily dose increased to either delapril 30 mg+indapamide 2.5 mg or captopril 50 mg+hydrochlorothiazide 25 mg tablets for a further 5 months. The primary assessment of antihypertensive efficacy was the percentage of patients who responded after a 6-month drug treatment. The responder rates were 72.6% with D+I and 62.9% with C+H (P=0.004 between treatments) after 60 days of treatment, and 92.6% in the D+I and 85.2% in the C+H (P<0.001 between treatments) at the end of the treatment period. The final value of systolic blood pressure was 134.5+/-13.1 mmHg with D+I and 138.3+/-14.0 mmHg with C+H (P<0.001 between treatments). At the final visit, DBP was 84.57+/-7.0 mmHg in the D+I group and 85.57+/-8.0 mmHg in the control group (P=0.017 between treatments). In all, 11 patients in the D+I group and 19 patients in the C+H group were withdrawn from the study because of adverse events. In all, 30 patients (7.6%) with D+I and 32 patients (8.1%) with C+H experienced adverse events. In conclusion, D+I was more effective than C+H in terms of overall reduction in blood pressure and response rate. Greater efficacy was obtained without any increase in adverse effects, since both treatments were equally well tolerated. PMID- 12574794 TI - Recovery of normal diurnal variation of blood pressure following renal artery stenting. AB - Renal artery stenting is a therapeutic option in patients with functionally significant atherosclerotic renovascular disease. Improved control of blood pressure (BP) and preservation of renal function may be achieved following intervention. We present a case in which renal artery stenting was followed by recovery of the normal diurnal variation in BP. PMID- 12574795 TI - Diuretic-induced hyponatraemia in elderly hypertensive women. PMID- 12574796 TI - Compression sonography for the exclusion of deep vein thrombosis: can I trust my eyes? PMID- 12574797 TI - Thrombinography shows acquired resistance to activated protein C in patients with lupus anticoagulants. AB - In patients with lupus anticoagulants (LA), acquired resistance to activated protein C (APC) is difficult to demonstrate with clot-based assays due to the presence of the anticoagulant. Via the conversion of a fluorogenic substrate (thrombinography), we monitored the complete process of thrombin formation and decay and its delimitation by the protein C system in eight consecutive LA patients without anticoagulant therapy and non-carriers of the V Leiden polymorphism. Thrombin generation was triggered in platelet-poor and platelet rich plasma by recalcification in the presence of a low concentration of tissue factor. In 7 out of 8 patients we observed a long lag-time before the thrombin burst (LA effect) together with a marked inability of APC to diminish the thrombin activity. The lag-phase was however prolonged to some degree by APC. The effects were more outspoken in the presence of phospholipids from patients' platelets than with added phospholipids. Thrombinography thus demonstrates APC resistance in LA-patients despite the occurrence of long lag-times (clotting times). The amount of thrombin activity generated in the presence of APC could be a better indicator of the thrombotic risk than the moment at which the thrombin burst starts. PMID- 12574798 TI - Molecular mechanisms of leukocyte recruitment: organ-specific mechanisms of action. AB - Leukocyte recruitment in the microcirculation is a critical process underlying inflammatory responses in tissues. The mechanism of recruitment is summarized by the generally accepted paradigm: selectin-mediated leukocyte rolling, subsequent integrin activation, followed by integrin-mediated firm adhesion. This simple paradigm may not be able to explain the leukocyte recruitment mechanisms in some organs including the liver and brain. Recent studies suggested that these organs have their own leukocyte recruitment paradigms in acute and even chronic inflammation. The combination of unique hemodynamic patterns and specific structural and functional features of the vessels and endothelium in liver may dictate and select the specific patterns of leukocyte recruitment in this organ. In the brain microvasculature, where shears are high and adhesion molecule expression low, platelets may play an important role as a bridge between the leukocytes and endothelium. PMID- 12574799 TI - A single complete ultrasound investigation of the venous network for the diagnostic management of patients with a clinically suspected first episode of deep venous thrombosis of the lower limbs. AB - In patients clinically suspected of deep-vein thrombosis (DVT) of the lower limbs, it is safe to withhold anticoagulant therapy after a negative ultrasound (US) limited to the popliteal and the femoral veins, provided that this can either be repeated or combined with other diagnostic procedures. To assess the safety of withholding anticoagulants after a single negative complete US, we performed a multicenter, prospective, cohort study including consecutive ambulatory outpatients from institutional and private practice settings, with a clinically suspected first episode of DVT. Patients fulfilling the inclusion criteria were enrolled after careful clinical assessment. A complete US examination of the proximal and the distal veins was performed according to a standardized and detailed protocol. Anticoagulant therapy was administered in patients with proximal or isolated distal DVT and withheld in those with negative results. The main outcome measure was the occurrence of objectively documented clinical thromboembolic events during a three-month follow-up after a negative US. Out of 623 patients, 401 (64.4%) had a baseline negative US, were not anticoagulated and could be followed-up for three months. Two patients presented a calf DVT within three months. The incidence of venous thromboembolic events, including distal DVT, was 0.5% [95% confidence interval: 0.1-1.8]. No proximal DVT, or non-fatal or fatal pulmonary embolism occurred (incidence: 0.0% [95% confidence interval: 0.0-0.9]). In conclusion, it is safe to withhold anticoagulant therapy in patients with clinically suspected DVT after a single, negative, complete US. Integrating this method within diagnostic strategies for DVT could improve management and be more acceptable for patients and physicians. PMID- 12574800 TI - Complete compression ultrasonography of the leg veins as a single test for the diagnosis of deep vein thrombosis. AB - Noninvasive diagnosis of deep vein thrombosis (DVT) is based on ultrasound examination of the leg veins, usually restricted to only compression of the proximal veins (CUS). Patients with negative CUS findings require a second examination or a combination with other tests, which impairs clinical efficiency. In this prospective outcome study, 1646 consecutive patients with clinically suspected DVT were examined once by a standardized protocol of complete compression ultrasound comprising all proximal and distal veins (CCUS) as the only diagnostic test. The examination was equivocal in 15 patients (1% technical failure rate). Another 366 patients (22%) were tested positive for proximal DVT, distal DVT, muscle vein thrombosis, or phlebitis. Of 1265 patients in whom CCUS findings were negative, 242 met exclusion criteria for follow-up (age <18, life expectancy <3 months, other reasons for anticoagulation, postthrombotic lesions of the leg veins, or lack of informed consent). During the 3 months of follow-up, three of 1023 patients with negative CCUS findings experienced a symptomatic venous thromboembolic event (0.3% [95% CI 0.1%-0.8%]). We conclude that the CCUS protocol has a low technical failure rate and is safe with respect to excluding DVT, thereby reducing the diagnostic workup of patients with suspected DVT to a single ultrasound examination. PMID- 12574801 TI - Effect of factor VIII on tissue factor-initiated spatial clot growth. AB - Using time-lapse videomicroscopy, we studied the role of coagulation factor VIII (fVIII) in tissue factor-initiated spatial clot growth on fibroblast monolayers in a thin layer of non-stirred recalcified plasma from healthy donors or patients with severe Haemophilia A. Analysis of temporal evolution of light-scattering profiles from a growing clot revealed existence of two phases in the clot growth initiation phase in a narrow (0.2 mm) zone adjacent to activator surface and elongation phase in plasma volume. While the initiation phase did not differ in normal and haemophilic plasmas, the rate of clot growth in the elongation phase in haemophilic plasma constituted only 30% of that in normal plasma. Supplementation of haemophilic plasma with 0.05 U/ml fVIII restored the normal clot growth rate (44.9 +/- 2.5 microm/min) at high but not at low fibroblast density. Our results indicate that the functioning of the intrinsic tenase complex is critical for normal spatial clot growth. PMID- 12574803 TI - Deficiency of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) impairs nutritionally induced obesity in mice. AB - Tissue inhibitor of matrix metalloproteinase-1 deficient (TIMP-1(-/-)) mice and wild-type (TIMP-1(+/+)) controls were kept on a standard (SFD) or a high fat diet (HFD) for 15 weeks. At the time of sacrifice, TIMP-1(-/-) mice on HFD had a significantly lower body weight (29 +/- 1.5 versus 41 +/- 1.8 g, p <0.005), and significantly less subcutaneous (0.81 +/- 0.19 versus 1.78 +/- 0.21 g, p <0.05) and gonadal (0.87 +/- 0.17 versus 1.85 +/- 0.18 g, p <0.005) fat mass. These differences were much less pronounced for mice on SFD. On HFD but not on SFD, adipocyte diameters were significantly lower in the adipose tissue of TIMP-1(-/-) mice. Plasma leptin levels in TIMP-1(-/-) mice on HFD were significantly lower as compared to TIMP-1(+/-) mice, and strongly correlated with adipose tissue mass for both genotypes. Staining with an endothelial cell specific lectin revealed a significantly higher blood vessel density, larger stained area and vessel size in adipose tissue of TIMP-1(-/-) mice on HFD. This difference disappeared after normalization to the adipocyte number, suggesting that it does not represent a true enhancement of angiogenesis. Thus, in a murine model of nutritionally induced obesity, TIMP-1 promotes adipose tissue development. PMID- 12574802 TI - Impaired prothrombinase activity of factor X Gly381Asp results in severe familial CRM+ FX deficiency. AB - We investigated three members of a large Omani family affected by severe factor X (FX) deficiency (coagulant activity <1%) and showing marked differences in the onset of severe hemorrhagic symptoms. All patients were homozygous for a novel FX mutation (Gly381Asp) in the structurally conserved region of the serine protease active site. Expression levels of recombinant 381D-FX were similar to those of wt FX, indicating the presence of a severe CRM+ FX deficiency, a poorly investigated condition. The 381D-FX was normally activated and did not show a detectable amidolytic activity. Instead, we observed a residual activity in a prothrombin time based assay (1%) and in prothrombinase assays both in plasma (1%) and in purified systems (3%). Comparison with FX variants characterized by reduced activation suggests that mutations affecting FX activity might result in a more pronounced impairment of coagulation and thus in severe hemorrhagic phenotype. In addition, this study indicates that the hemorrhagic heterogeneity observed in FX deficiencies is only partially explained by molecular analysis of FX gene. PMID- 12574804 TI - In vivo pig models of venous thrombosis mimicking human disease. AB - Most animal models of venous thrombosis involve acute thrombosis with hypercoagulability in small rodents. To better replicate human disease, we developed two models in the pig, a species similar to humans in size and in vascular and coagulation reactivity. One model involves de-endothelialisation with 50% or 80% stenosis and the other replacement of a venous segment by a Gore Tex vascular prosthesis. Both models were tested with and without acute induced hypercoagulability (thromboplastin infusion). Thrombi obtained without thromboplastin infusion were composed of a multilayered platelet and a fibrin meshwork structure similar to that usually found in humans. With thromboplastin infusion, the thrombi were homogeneous fibrin structures imprisoning red blood cells. The high incidence of thrombosis obtained with the 80% stenosis model would be useful for studying anticoagulant treatments, whereas the low incidence with 50% stenosis would be useful for evaluating procoagulant effects of conditions or treatments. These new models shed further light on the development of venous thrombi under conditions similar to those seen in humans and may prove useful for investigating anticoagulant and procoagulant effects. PMID- 12574805 TI - Different mechanisms contribute to the biphasic pattern of carboxypeptidase U (TAFIa) generation during in vitro clot lysis in human plasma. AB - Carboxypeptidase U (CPU,TAFIa) recently gained interest as a significant player in dampening the fibrinolytic rate. The aim of this study was to investigate the time course of the generation of CPU activity during coagulation and fibrinolysis using an in vitro clot lysis model in human plasma. A first peak of CPU activity appeared after initiation of the coagulation phase and a second rise in CPU activity was observed during the fibrinolysis. The decrease in the proCPU plasma concentration followed the same trend as the appearance of the CPU activity. The direct thrombin inhibitor inogatran eliminated the CPU generation during coagulation but not during fibrinolysis. Addition of the plasmin inhibitor aprotinin during fibrinolysis resulted in a decrease in CPU activation during the lysis phase. These results demonstrate that proCPU was activated during coagulation by thrombin and during fibrinolysis by plasmin. Addition of a CPU inhibitor before initiation of clotting decreased the clot lysis time as expected. However, addition in the time period between the two peaks of CPU activity had no apparent effect on the clot lysis time. PMID- 12574806 TI - Risk factors and coagulation parameters in relationship to phlebographic response and clinical outcome in the treatment of acute deep vein thrombosis. AB - Possible correlation of the effects of pharmacotherapy on the inhibition of the in-vivo generation of thrombin and on the prevention of thrombus extension in patients with deep vein thrombosis (DVT) could help to define patients at higher risk. Patients with symptomatic deep vein thrombosis confirmed by phlebography were randomised to intravenous unfractionated heparin (UFH), or a subcutaneous low-molecular-weight heparin (reviparin) twice daily for one week, or a subcutaneous reviparin once daily for four weeks. The patients were treated with oral anticoagulants for at least 3 months. Main endpoints were regression of thrombus on phlebography on Day 21 and recurrent symptomatic venous thromboembolism up to 3 months. Coagulation parameters, markers of in-vivo thrombin generation, and TFPI-release were determined at randomisation, weeks 1 and 3. Four hundred sixty six responders (reduction of at least 30 per cent in Marder score) and 419 non-responders (Marder score unchanged or changed less than +/-30%) showed no significantly different baseline characteristics. The non responder group had a higher median Marder score at baseline and after one and three weeks of treatment, and had significantly higher fibrinogen levels, TAT complexes and F1+2 values than responders. There were no significant differences in coagulation parameters between non-responders and patients with asymptomatic + symptomatic VTE with the exception of higher TAT complexes at baseline. Significant differences in Marder score and coagulation parameters at baseline were found between responders and nonresponders. Non-responders have a higher risk tosuffer recurrent VTE and may need intensified treatment. PMID- 12574807 TI - Predisposing factors for enlargement of intracerebral hemorrhage in patients treated with warfarin. AB - To elucidate predisposing factors for enlargement of intracerebral hematoma (ICH) during warfarin therapy, we reviewed 47 patients on warfarin who developed acute ICH and determined relationships among ICH enlargement, INR reversal and clinical data. Among 36 patients treated to counteract the effects of warfarin within 24 h of onset, ICH increased in 10 patients (enlarged group), but remained unchanged in the remaining 26 (unchanged group), while ICH remained unchanged in another 11 patients in whom the effect of warfarin was reversed after 24 h. The international normalized ratio (INR) was counteracted immediately in 11 patients treated with prothrombin complex concentrate (PCC) but gradually in the other 36 treated by reducing the dose of warfarin, or by administering vitamin K or fresh frozen plasma. Multivariate analysis with a logistic regression model showed an INR value <2.0 at admission or for 24 h after immediate INR correction with PCC prevented ICH enlargement (OR 0.069, 95%CI 0.006-0.789, p = 0.031). An INR value of >2.0 within 24 h of ICH seems an important predisposing factor for ICH enlargement. PMID- 12574808 TI - Prospective assessment of the natural history of positive D-dimer results in persons with acute venous thromboembolism (DVT or PE). AB - The natural history of initially positive D-dimers for venous thromboembolism is not known. If it returns to negative in the majority of patients, it would be potentially helpful to diagnose a recurrence. In this study, we prospectively measured D-dimer levels in outpatients with a diagnosis of venous thromboembolism. There were a total of 152 patients with an average age of 57. D dimer results were performed at baseline and repeated at one week, one month and three months. At baseline 120 of 152 (79%) had a positive D-dimer result. Of those with an initially positive result, 80% were still positive at one week and 39% were still positive at one month. Finally at three months, 13% remained positive. Seven patients had recurrent events and all had persistently elevated D dimers at one month. This study suggests that a persistently positive D-dimer result after one month of treatment may indicate a higher risk of recurrent venous thromboembolism. D-dimer testing for the diagnosis of recurrence of venous thromboembolism deserves further study. PMID- 12574809 TI - Direct thrombin inhibitor melagatran followed by oral ximelagatran in comparison with enoxaparin for prevention of venous thromboembolism after total hip or knee replacement. AB - We evaluated whether a postoperative regimen with melagatran followed by oral ximelagatran, two new direct thrombin inhibitors, was an optimal regimen for thromboprophylaxis in major orthopaedic surgery. In a double-blind study, 2788 patients undergoing total hip or knee replacement were randomly assigned to receive for 8 to 11 days either 3 mg of subcutaneous melagatran started 4-12 h postoperatively, followed by 24 mg of oral ximelagatran twice-daily or 40 mg of subcutaneous enoxaparin once-daily, started 12 h preoperatively. Ximelagatran was to be initiated within the first two postoperative days. The primary efficacy endpoint was venous thromboembolism (deep-vein thrombosis detected by mandatory venography, pulmonary embolism or unexplained death). The main safety endpoint was bleeding. Venous thromboembolism occurred in 355/1146 (31.0%) and 306/1122 (27.3%) patients in the ximelagatran and enoxaparin group, respectively, a difference in risk of 3.7% in favour of enoxaparin (p = 0.053). Bleeding was comparable between the two groups. PMID- 12574810 TI - A randomized trial of initial warfarin dosing based on simple clinical criteria. AB - Warfarin induction is accomplished by titrating dosage to coagulation test results. Algorithms can guide this process but not identify the starting dose. We hypothesized that an initial warfarin dose approximating the maintenance value would safely enhance rapidity of induction. In a randomized trial we compared a fixed-dose to a maintenance-dose strategy for beginning warfarin therapy. To predict the maintenance dose among patients with differing warfarin requirements we performed regression analysis on clinical factors derived from chart review. Four community hospitals supplied records for retrospective analysis. The prospective trial was conducted in one, a 350-bed teaching institution. A sample of inpatients anticoagulated during 1998 formed the development set for retrospective study; a 1999 sample formed the validation set. A one year trial recruited consecutive eligible inpatients initiated on warfarin. We randomly assigned patients to a first warfarin dose calculated using our regression formula or fixed at 5 mg. All patients' subsequent doses were determined (as a percentage of initial) from coagulation testing. We compared days to anticoagulation, hospitalized hours, complications, and activity of factor II and protein C in a patient sample at intervals after induction. Weight, age, serum albumin, and presence of malignancy explained 25-30% of variance in maintenance dose. Ninety patients (44 calculated-dose and 46 standard-dose) evaluated in the clinical trial. Mean time to anticoagulation (among patients achieving anticoagulation) was 4.2 and 5.0 days, respectively (p = 0.007). We observed no significant differences in other endpoints. Individualized initial dosing may safely hasten warfarin induction. PMID- 12574811 TI - Body mass index is associated with the development of the post-thrombotic syndrome. AB - Post-thrombotic syndrome (PTS) is a chronic complication of deep vein thrombosis (DVT). Little is known about prognostic factors that might identify patients at high risk for the development of PTS. Body mass index (BMI) has been previously reported to be associated to the development of PTS. The aim of this study was to assess the association between BMI and other anthropometric parameters and PTS in a general population of DVT patients. In a prospective cohort study, 83 consecutive patients with objective diagnosis of DVT underwent physical examination. BMI was recorded at baseline and at 12 months, and waist circumference was recorded at 12 months to assess individual patterns of body fat distribution. The presence of PTS at 12 months was ascertained using a validated clinical scale. Sixty-three patients (75.9%) were overweight or obese at 12 months, 60 (72.3%) had a weight gain over 1 year. Twenty patients developed PTS (24.1%). Mean BMI was significantly higher in patients who developed PTS than in patients who did not (29.6 and 27.2 Kg/m(2), respectively, p = 0.022). A BMI of > 28 Kg/m(2) predicted early onset of PTS (OR 3.54, 95% CI 1.07-12.08, p = 0.017). Neither patterns of fat distribution nor weight gain in 1 year were correlated with PTS (p = 0.918 and p = 0.775, respectively). BMI is significantly correlated with the development of PTS. Patients with DVT should be encouraged to avoid weight gain. Reducing patient weight might be an important strategy to prevent PTS. PMID- 12574812 TI - Differential regulation of P-selectin expression by protein kinase A and protein kinase G in thrombin-stimulated human platelets. AB - P-selectin is rapidly translocated from platelet alpha-granules following activation. Intracellular cyclic AMP (cAMP) is a potent inhibitory pathway that results in global downregulation of platelet activation. While cAMP-dependent protein kinase (PKA) has long been considered as the main mediator of cAMP dependent effects, no study has yet evaluated its effect on P-selectin expression in human platelets. Pretreatment of thrombin-stimulated platelets with forskolin resulted in a concentration- dependent inhibition of P-selectin expression that correlated with adenylyl cyclase activity. Inhibition of PKA with H-89 reversed cAMP-induced inhibition of P-selectin while cGMP-dependent protein kinase (PKG) inhibition with KT5823 significantly potentiated cAMP-dependent inhibition of P selectin. Similar results were also observed in a platelet/neutrophil binding assay. In conclusion, cAMP-induced inhibition of P-selectin expression is, in large part, mediated through activation of PKA. PKG appears to be solicited for P selectin expression when cAMP levels are elevated which suggest a cAMP/PKG dependent pathway of platelet activation. PMID- 12574813 TI - A critical role of lipid rafts in the organization of a key FcgammaRIIa-mediated signaling pathway in human platelets. AB - The involvement of platelet FcgammaRIIa in heparin-associated thrombocytopenia (HIT) is now well established. However, the precise sequence of molecular events initiated by FcgammaRIIa cross-linking in platelets remains partly characterized. We investigated here the role of lipid rafts in the spatio-temporal organization of the FcgammaRIIa-dependent signaling events. Upon cross-linking, FcgammaRIIa relocated in rafts where the kinase Lyn and the adapter LAT were among the major phosphotyrosyl proteins. Upon stimulation by HIT sera, the second messenger phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) accumulated in rafts in a P(2)Y(12) adenosine diphosphate (ADP) receptor-dependent manner. PtdIns(3,4,5)P(3) was then essential to specifically recruit phospholipase Cgamma2 (PLCgamma2) to these membrane microdomains. Controlled disruption of rafts by methyl beta-cyclodextrin reversibly abolished PtdIns(3,4,5)P(3) production, PLC activation and platelet responses induced by FcgammaRIIa cross linking without affecting the tyrosine phosphorylation events. This work demonstrates that platelet rafts are essential for the integration of a key signaling complex leading to the rapid production of PtdIns(3,4,5)P(3) and in turn PLCgamma2 activation during HIT. PMID- 12574814 TI - Activation-dependent surface expression of gC1qR/p33 on human blood platelets. AB - GC1qR/p33 (gC1qR) is expressed by a variety of somatic and cultured cells, including blood platelets. It interacts with several cellular, viral, bacterial, and plasma proteins, suggesting a potential role in thrombosis, inflammation, and infection. Considerable controversy has surrounded the surface membrane localization of gC1qR, however, since its cDNA sequence does not predict a traditional membrane-anchoring domain, and bears a typical mitochondrial targeting sequence. The present study examined gC1qR expression on resting and activated human blood platelets using flow cytometry and confocal microscopy with two monoclonal antibodies, 74.5.2 and 60.11, directed against gC1qR C-terminal amino acids 204-218, and N-terminal amino acids 76-93, respectively. Unstimulated platelets reacted minimally with either antibody. In contrast, platelet activation with TRAP, epinephrine, or ADP produced markedly increased gC1qR expression as reflected by 74.5.2 binding but not 60.11 binding. Platelet activation was verified using PAC-1 and anti CD 62 antibodies. Whereas PAC-1 binding to activated platelets could be reversed following platelet incubation with PGE1, 74.5.2 binding remained unchanged, suggesting the sustained expression of gC1qR following platelet stimulation. The data further demonstrate that detection of cell surface gC1qR may be dependent on antibody specificity. The ability of gC1qR to bind proteins involved in complement, coagulation, and kinin systems, as well as viral and bacterial pathogens including S. aureus protein A, supports the hypothesis that gC1qR expressed on activated platelets may contribute directly to thrombosis, inflammation, and endovascular infections. PMID- 12574815 TI - Activation of human platelets by 2-arachidonoylglycerol is enhanced by serotonin. AB - The endocannabinoid 2-arachidonoylglycerol (2-AG) has been shown to activate human platelets in platelet-rich plasma, by binding to a "platelet-type" cannabinoid receptor (CB(PT)). Here, washed human platelets were used to characterize the binding of [(3)H]2-AG to CB(PT), showing a dissociation constant (Kd) of 140 +/- 31 nM and a maximum binding (Bmax) of 122 +/- 10 pmol.mg protein( 1). Selective antagonists of both CB1 and CB2 cannabinoid receptors inhibited this binding, which was enhanced up to approximately 230% over the controls by 1 micro M serotonin (5-hydroxytryptamine, 5-HT). Human platelets were also able to bind [(3)H]5-HT (Kd = 79 +/- 17 nM, Bmax = 14.6 +/- 1.3 pmol.mg protein(-1)), and 1 micro M 2-AG enhanced this binding up to approximately 150%. Moreover, they were able to take up [(3)H]5-HT through a high affinity transporter (Michaelis Menten constant = 22 +/- 2 nM, maximum velocity = 344 +/- 15 pmol.min(-1).mg protein(-1)), which was not affected by 2-AG. Interestingly, 5-HT did not affect the activity of the 2-AG transporter of human platelets. Treatment of washed platelets with 1 micro M 2-AG led to increased intracellular inositol-1,4,5 trisphosphate (up to approximately 300%) and decreased cyclic AMP (down to approximately 50%). Furthermore, treatment of pre-loaded platelets with 1 micro M 2-AG induced a approximately 300% increase in [(3)H]2-AG release, according to a CBPT-dependent mechanism. Also, 1 micro M 5-HT enhanced the effect of 2-AG on inositol-1,4,5-trisphosphate ( approximately 500% of the controls), cyclic AMP ( approximately 20%) and [(3)H]2-AG release ( approximately 570%), and the latter process was shown to be partly ( approximately 50%) involved in the 5-HT dependent platelet activation. Taken together, reported findings represent the first demonstration that 2-AG and 5-HT can mutually reinforce their receptor binding on platelet surface, which might have therapeutic implications. PMID- 12574816 TI - Effect of fibrinogen concentration and platelet count on the inhibitory effect of abciximab and tirofiban. AB - Glycoprotein (Gp) IIb/IIIa blockers are powerful antithrombotic agents, but display a wide variability of their effect using weight adjusted dosing. We investigated whether some hemostatic variables affected in vitro platelet inhibition exerted by the antibody abciximab and the peptidomimetic tirofiban. High fibrinogen concentrations reduced platelet inhibition by tirofiban, assessed in whole blood (r = -0.85, n = 10, p <0.01) and in platelet rich plasma (r = 0.89, n = 10, p <0.01) through PFA 100 and Born's aggregometry assay, respectively. Both drugs were unaffected by von Willebrand factor levels, while platelet count was inversely related to their inhibitory effect (tirofiban: r = 0.9, n = 7, p <0.01; abciximab: r = -0.81, n = 9, p <0.01). Analysis of GpIIb IIIa blockade showed that receptor occupancy at a fixed abciximab dose was inversely related to platelet counts. These experimental data were in agreement with the classical model of receptor saturation by a tight binding inhibitor. A role for fibrinogen and/or platelet count in influencing the antithrombotic properties of platelet GpIIb-IIIa antagonists can be hypothesized. PMID- 12574817 TI - Blood coagulation and fibrinolytic factors in 105 patients with hemorrhagic syndrome caused by accidental contact with Lonomia obliqua caterpillar in Santa Catarina, southern Brazil. AB - Haemostatic disorders caused by Lonomia obliqua caterpillars has reached epidemic proportions in southern Brazil. Here we evaluated coagulation and fibrinolysis in 105 patients after accidental contact with Lonomia obliqua caterpillars. Global coagulation tests were prolonged in most cases and patients were divided into 3 groups according to fibrinogen (Fg) level: 1.5 g/l (group C). There was a significant reduction of factors V, XIII, VIII and prekallikrein in group A, with no change in factors X, II and von Willebrand factor. Thrombin-antithrombin and prothrombin F1+2 were elevated in most patients. Antithrombin and protein S were not changed whereas protein C levels were reduced in group A. Plasminogen and alfa2-antiplasmin levels were significantly reduced in group A and D-Dimer levels were extremely high in all groups, showing that fibrinolysis had been activated, possibly secondary to fibrin production. Levels of t-PA were normal and PAI-1 was mildly elevated in group A. The platelet count remained above 150 x 109 platelets/ml in 97% of cases. In summary, our results suggest that Lonomia obliqua envenoming is characterized by a consumption coagulopathy and secondary fibrinolysis. PMID- 12574818 TI - Effect of clotting factors concentrates on lymphocyte and neutrophil function in vitro. AB - Immunological abnormalities have been reported in haemophiliacs. Although infections with HIV, hepatitis and other viruses may contribute to these abnormalities, immune defects are detectable also in HIV seronegative haemophiliacs. It is likely that chronic exposure to extraneous proteins in clotting factor concentrates (CFCs) may play a role in immunomodulation, but the underlying mechanisms remain unclear. The results of the present paper show that: a) soluble HLA class I (sHLA-I), soluble Fas-ligand (sFas-L) and transforming growth factor beta 1 (TGF-beta1) are detectable in plasma derived but not in recombinant CFCs; b) the level of sHLA-I and sFas-L is proportional to the grade of CFCs purity whereas TGF-beta1 showed very variable levels; c) soluble molecules detected in CFCs exert immunomodulatory effects in vitro like apoptosis induction in Jurkat cells and inhibition of mixed lymphocyte reaction response, antigen-specific lymphocyte cytotoxic activity and neutrophil chemotaxis. PMID- 12574819 TI - Fibrinolytic capacity increases with age in healthy humans, while endothelium dependent vasodilation is unaffected. AB - The capacity for stimulated endothelial release of tissue-type plasminogen activator (tPA) and endothelium-dependent vasodilation is diminished in patients with cardiovascular risk factors. We examined the effect of age on desmopressin stimulated tPA release and endothelium-dependent vasodilation, using the perfused forearm model. Thirty-two healthy subjects were divided into quartiles by age (mean age 24, 36, 54, and 72 years, respectively). Baseline fibrinolytic parameters, baseline forearm blood flow (FBF), and increase in stimulated FBF were all similar across age. By contrast, the capacity for desmopressin- stimulated release of tPA increased linearly by age. For tPA antigen, the total amount released (area-under-the-curve) was 1015, 1282, 2139, and 2845 ng/L tissue (p = 0.011) and the peak release rates were 62, 80, 113, and 163 ng/min/L tissue (p = 0.008) in the age-quartiles, respectively. A similar significant age-related response was observed for tPA activity. We conclude that in healthy individuals there is an up-regulation of the fibrinolytic response by age. PMID- 12574820 TI - Human/chicken urokinase chimeras demonstrate sequences outside the serine protease domain that dictate autoactivation. AB - Mammalian urokinase-type plasminogen activator (uPA) is produced as a stable single polypeptide chain zymogen and requires a distinct proteolytic cleavage to become an active, two-chain enzyme. In contrast, chicken uPA, both native and recombinant, is found predominantly as a two-chain, active enzyme even in the absence of plasmin, a physiological activator. Here we show that the proclivity to autoactivate is not a unique property of the chicken uPA catalytic domain but requires sequences distinct from and independent of the serine protease domain. Human/chicken chimeric uPA molecules and point mutants were used to determine the structural requirements for uPA autoactivation versus zymogen stability. The amino terminal fragment of chicken uPA engineered onto the human uPA molecule can induce the autoactivation of the human uPA. In fact, the first twenty residues of the chicken uPA are necessary and sufficient to induce the autoactivation of chicken and human uPA. These results indicate that sequence motifs, distal to the active site, control the substrate specificity and catalytic efficiency of uPA activity in autolytic activation. PMID- 12574821 TI - Aging of stromal-derived human breast fibroblasts might contribute to breast cancer progression. AB - Age is an important factor in the development and spread of breast cancer. Stromal cells also contribute to breast cancer growth and metastasis through the production of extracellular matrix (ECM) modifiers such as urokinase type plasminogen activator (uPA), its receptor (uPAR), its inhibitors (PAI-1 and PAI 2), matrix metalloproteinases (MMPs), and growth factors, including the fibroblast and insulin-like growth factors (FGF's and IGF's). In the present study we have investigated whether breast fibroblasts aged in vitro through passage in culture display altered levels of the plasminogen activator system and growth factors that are known to modulate that system. With real-time RT-PCR we found that during passage human breast fibroblasts, whether derived from the tumour burden or from matched adjacent normal breast tissue, exhibited a consistent increase in PAI-1 and FGF-1 and a decrease in MMP-2 mRNA expression. In addition, in 5 out of 7 fibroblast strains we observed an induction of uPA expression in combination with a reduced IGF-1 expression. Interestingly, while during aging MMP-2 protein increased in all tumour-derived fibroblast strains, these protein levels were reduced in all normal tissue- derived fibroblasts. No other clear-cut age-dependent alterations were found in the all-together 25 factors investigated. We furthermore demonstrate in one tumour-derived fibroblast strain that the increases in uPA and PAI-1 mRNA and MMP-2 protein production are inversely related to the telomere length. Artificially increasing telomere length in this fibroblast strain by expressing human telomerase reverse transcriptase (hTERT) prevented senescence and resulted in late passage cultures with early passage uPA, PAI-1 and MMP-2 levels. Our results show that aging accompanied by telomere loss induces PAI-1 and FGF-1 mRNA expression in all breast fibroblast strains, increases uPA and decreases IGF-1 mRNA expression in a subset, and increases MMP-2 protein expression only in tumour-derived breast fibroblasts. These age-induced levels of PAI-1, FGF-1, uPA and MMP-2 in stromal breast fibroblast could contribute to breast cancer progression. PMID- 12574822 TI - Low molecular weight heparin treatment for malignant melanoma: a pilot clinical trial. PMID- 12574823 TI - A reply to rebuttal: What is the risk of withholding anticoagulation in those with a low probability ventilation perfusion scan in the investigation of pulmonary emboli? PMID- 12574840 TI - The histopathology of pediatric celiac disease: order must prevail out of chaos. AB - The role of histopathology for diagnosing celiac disease (CD) has been recently challenged. However, based in our experience with roughly 4,600 distal duodenal and jejunal biopsies in children it is apparent that appropriate biopsy site, handling, processing, and microscopic evaluation result in a consistent pattern of microscopic changes which allows strong clinical-pathologic correlation. A simple way for establishing the villous/crypt (V/C) ratio is proposed. Normal mucosa displays a V/C ratio of 2.5 or more. Villous atrophy is then graded according to the V/C ratio as follows: Grade 1: 2.5-2; Grade 2: 1-2; Grade 3: 1 0.5, and Grade 4: less than 0.5. The grading should be done in areas of the biopsy where at least 2 to 3 crypts are present in almost its full length. CD disease was consistently associated with villous atrophy grades 3 and 4, which fully recovered or maintained Grade 1 after gluten-free diet. Grade 2 biopsies were rare and related to incomplete gluten-free diet. Patchy lesions were never seen as were patients with normal biopsies later developing mucosal atrophy. Histopathologic evaluation of mucosal biopsies to rule out CD requires adequate biopsy site (distal duodenum or proximal jejunum), and proper handling (oriented material), processing (cutting on edge) and interpretation. The proposed villous atrophy grading may help to adequately compare experiences from different centers as well as to reconcile apparent different findings in separate biopsies. In children histopathology keeps on having a central role for CD diagnosis. PMID- 12574841 TI - Real-time quantitative analysis of E-cadherin expression in ret/PTC-1-activated thyroid neoplasms. AB - Papillary thyroid carcinoma (PTC), the most common variety of thyroid cancer, is found in a variety of morphologic variants, usually grows slowly, and is clinically indolent, although rare, aggressive forms, with local invasion or distant metastases, occur. Our group has previously demonstrated an association between Hashimoto thyroiditis and ret/PTC-1 activation, and have hypothesised that c-ret activation might be implicated in immune reaction to thyroid epithelium. The objective of this study was to examine expression of the cellular adhesion molecule, E-cadherin, in various thyroid tumor types and Hashimoto thyroiditis in the context of ret/PTC-1 positivity by using laser capture microdissection and TaqMan reverse transcription-polymerase chain reaction (RT PCR). Variable down-regulation of E-cadherin among carcinomas was demonstrated, with anaplastic carcinomas showing little or no expression. Follicular thyroid carcinomas consistently had significantly decreased E-cadherin expression compared with papillary thyroid carcinomas. The ret/PTC-1-positive papillary thyroid carcinoma (PTCret+) and Hashimoto thyroiditis cases had consistently lower E-cadherin expression levels than the corresponding ret/PTC-1-negative papillary carcinomas (PTCret-), suggesting not only an association between ret activation and the loss of cellular adhesion but also, more significantly, an association between papillary thyroid carcinoma and Hashimoto thyroiditis. PMID- 12574842 TI - Nuclear localization of beta-catenin and plakoglobin in primary and metastatic human colonic carcinomas, colonic adenomas, and normal colon. AB - Beta-catenin is a cytoskeleton-associated signaling molecule shown to be elevated in various carcinomas but mostly in colon cancer owing to its impaired degradation. In contrast, its close homologue plakoglobin was shown to suppress the tumorigenicity of certain tumor cells. In the present study, we have used a semiquantitative immunohistochemical approach to evaluate the extent of nuclear localization of beta-catenin in human colonic adenocarcinomas and adenomas and compared it to the distribution of plakoglobin in the same tissues. We show that beta-catenin accumulates in the nuclei of the epithelium of primary and metastatic colonic adenocarcinoma as well as in colonic adenomas. In contrast, nuclear plakoglobin levels in these tissues were low, even compared to those found in epithelial cells of normal colon. These results support the view that the increase in beta-catenin levels in colon cancer cells occurs early in the tumorigenic process, leading to its nuclear localization, not only in invasive adenocarcinoma, but also in colonic adenoma with mild dysplasia. PMID- 12574843 TI - Tenascin and strictures in inflammatory bowel disease: an immunohistochemical study. AB - Tenascin is an extracellular matrix protein involved in morphogenesis of muscle tissue and in wound healing. In the present study we examined its distribution in tissue from patients with inflammatory bowel disease. Intestinal biopsies from 10 normal controls, 15 patients with Crohn's disease, and 6 with ulcerative colitis were studied. Samples were obtained both from uninvolved and involved areas. Mucosal tenascin is increased in ulcerative colitis and Crohn's disease, especially in areas of ulceration. In Crohn's disease, tenascin is also strongly expressed in the submucosa and in smooth muscle cells of the muscularis mucosae and propria, especially in areas of stricture. We conclude that tenascin is involved in stricture formation in Crohn's disease and that it is a marker of phenotypic change in smooth muscle cells. PMID- 12574844 TI - Follicular lymphoma of the salivary gland: a clinicopathological and molecular study of six cases. AB - To clarify the clinicopathologic, immunohistologic, and genotypic features of follicular lymphoma arising from the salivary glands, we examined 20 cases of operatively resected primary salivary gland lymphoma and identified 6 such cases. There were 4 women and 2 men with ages ranging from 38 to 64 years (median 50 years). The tumor arose from the parotid gland in 4 cases and the submandibular gland in the remaining 2. Four patients were stage IE and 2 were stage IIE-1. The median follow-up period was 49 months and all patients were alive and well at the time of going to press. Histologically, 5 patients were follicular lymphoma grade 2, and 1 was grade 3. In all specimens in noninfiltrating salivary gland tissue, there was periductal lymphocytic infiltration near the lymphoma. Moreover, myoepithelial sialoadenitis was noted in 2 lesions. An immunohistochemical study revealed all 6 cases were CD10+, CD79a+, bcl-6+, CD3-, CD5-, CD21-, CD23-, and CyclinD1-. The tumor cells expressed bcl-2 in 3 cases and p53 oncoprotein in 4 cases. Two cases revealed clonal bands with polymerase chain reaction (PCR) assay for the immunoglobulin heavy (IgH) gene. The bcl-2/IgH translocation at the major breakpoint region was detected in 1 case (16%). We found a relatively high incidence of follicular lymphomas (30%) in salivary gland lymphomas. Among the mucosa-associated lymphoid tissue (MALT) system, follicular lymphomas appeared to occur frequently in the salivary glands as well as the duodenum and skin. Moreover, follicular lymphoma arising from the salivary glands appeared to have some of the characteristics of MALT-type lymphoma including indolent prognosis, presence of myoepithelial sialoadenitis, and rarity of the BCL-2 gene rearrangement. PMID- 12574845 TI - Orbital hemangiopericytoma and solitary fibrous tumor: a morphologic continuum. AB - Solitary fibrous tumors (SFT) and hemangiopericytomas (HPC) are soft tissue tumors with known histologic and immunohistochemical overlap. A series of these tumors located in the orbit were analyzed in order to determine whether they could be re-classified based on currently recognized histologic criteria. Ten orbital spindle cell lesions, all of which were positive for CD34 antigen, were examined. Diagnostic criteria for SFT included a cytologically bland spindle cell lesion with variable cellularity and focal dense collagenization with diffuse, strong CD34 reactivity, while the criteria for HPC required a more monotonous cellular proliferation without significant variability in cellularity, a "staghorn" vascular pattern, minimal collagenization, and focal or absent CD34 staining. Tumors with typical histologic and immunohistochemical features of HPC or SFT were diagnosed as HPC and SFT, respectively. Those tumors with histologic or antigenic profiles not classic for HPC or SFT were defined as 'indeterminate.' Three lesions were classified as SFT and 1 tumor was diagnosed as HPC through use of the above-cited histologic criteria. All lesions showed positive staining of tumor cells with CD34 antigen in varying amounts and were negative for cytokeratin AE1-3, epithelial membrane antigen, CD68, and Factor XIIIa. One solitary fibrous tumor focally stained for S-100 protein and 1 hemangiopericytoma was focally positive for HHF-35. Of the 10 analyzed tumors, 6 were classified as 'indeterminate.' Furthermore, 1 lesion whose primary histology was that of an SFT recurred 9 years later with an appearance consistent with an 'indeterminate' lesion. Our results call into question the present histologic separation of HPC and SFT in the orbit. As in other sites, including deep soft tissue, these data suggest that SFT and HPC are 2 lesions whose morphologic features are best interpreted to exist along a continuum, rather than 2 lesions with distinctly defined histopathology. PMID- 12574846 TI - Images in pathology: myxoglobulosis of the appendix. PMID- 12574848 TI - Diagnostic pitfalls in surgical pathology--discovered by a review of malpractice claims: Part VII. Random errors. PMID- 12574849 TI - Monotypic angiomyolipoma of the nasal cavity: a heretofore undescribed occurrence. AB - A monotypic angiomyolipoma of the nasal cavity in a 34-year-old woman is described. Tumor cells were spindled or epithelioid and contained glycogen and diastase-resistant PAS-positive granules. There were few mitoses, and necrosis was absent, indicating a benign tumor. The stroma was markedly vascular, and a few adipocytes were seen in one area. Cells were positive for melanocyte and muscle markers. Electron microscopy revealed abundant dense granules. Although melanin was absent histochemically, it was present using a chemical assay, and the granules may, therefore, be atypical melanosomes. Fine actin filaments, attachment plaques and lamina were present. Initial assessment of the lesion indicated malignant melanoma, but the immunostaining and histologic features indicated monotypic angiomyolipoma. To the best of our knowledge, this is the first such case in the nasal cavity. PMID- 12574850 TI - Borderline serous cystadenocarcinoma with coexistent angiosarcoma: an unusual form of ovarian carcinosarcoma. AB - A 45-year-old woman presented with a short history of abdominal distension and features of intestinal obstruction. Computed tomography examination of the abdomen revealed a solid and cystic mass in the region of the left ovary, but spread within the abdomen was also noted. The ovarian tumor was removed, and an omentectomy was also performed. Microscopic examination of the ovarian tumor and metastases within the omentum showed a carcinosarcoma composed of borderline serous papillary cystadenocarcinoma and angiosarcoma. The angiosarcomatous component was the dominant tumor within the omentum. This case documents an unusual combination of epithelial and sarcomatous elements in an ovarian carcinosarcoma. The tumor behaved aggressively because of the angiosarcomatous component, and since the patient was assessed as having stage IV disease, no further treatment was offered. PMID- 12574851 TI - Recurrent chondromyxoid fibroma of the thoracic spine 30 years after primary excision: case report and review of the literature. AB - We report a case of late recurrence of chondromyxoid fibroma (CMF) arising in a thoracic vertebra in an 11-year-old male. This was treated by curettage, and 30 years later, the patient noticed shoulder pain and leg weakness. A recurrent mass appeared at the same site in the spinous process of T6. The histologic features of the recurrent tumor were similar to those of the primary lesion. A total of 38 cases of CMF of the vertebra have been reported. Only 3 of 38 previously reported vertebral CMF recurred. Tumors recurred 2 years after operation in 2 cases, and 7 years after operation in 1 case. PMID- 12574852 TI - Mammary epithelioid myofibroblastoma arising in bilateral gynecomastia: case report with immunohistochemical profile. AB - Myofibroblastoma of the breast is a rare benign neoplasm, which has rarely been reported in association with gynecomastia. We report a case of a 25-year-old male patient with an epithelioid myofibroblastoma arising in a context of bilateral gynecomastia. The lesion was composed of nests and cords of epithelioid cells, with round to oval nuclei, granular chromatin, and distinct nucleoli dispersed in a myxoid to collagenous stroma with marked vascular proliferation. Immunohistochemical profile showed diffuse positivity for vimentin and focal immunoreactivity for desmin, whereas cytokeratins (CAM5.2 and AE1/AE3), EMA, alpha smooth muscle actin, actin HHF35, CEA, S100, factor VIII, neuron-specific enolase, CD31, and CD34 were all negative. We emphasize that this diagnosis is difficult to establish, owing to the rarity of this variant and clinical presentation. PMID- 12574853 TI - Primary synovial sarcoma of the kidney: a case report with literature review. AB - We describe a case of primary renal synovial sarcoma (SS) in a 48-year-old man. The patient presented with hematuria and was found to have a large tumor in his left kidney on computed tomography scan. Histology revealed a highly cellular spindle cell neoplasm with minimal pleomorphism. The major differential diagnoses included leiomyosarcoma, hemangiopericytoma, and SS. The presence of focal areas with a biphasic pattern, uniformly positive immunostain for bcl-2, focally positive immunostains for epithelial membrane antigen and cytokeratin, and negative immunostains for CD-34, smooth muscle actin and S-100 established the diagnosis. This was subsequently confirmed by molecular testing for t(X;18) translocation. Since the existence of primary SS in the kidney was first suggested in 1999, to the best of our knowledge a total of 19 cases including the present case have been reported to date. Although primary renal SS is rare, these findings indicate that it should be included in the differential diagnosis of spindle cell tumors of the kidney. PMID- 12574854 TI - HMB45+ hyalinized epithelioid tumor of the uterus is linked to epithelioid leiomyoma rather than to PEC-omas. PMID- 12574855 TI - Carcinoma arising in microglandular adenosis of the breast. PMID- 12574856 TI - Analogies in medicine. PMID- 12574858 TI - Phylogeny and the evolution of the Amylase multigenes in the Drosophila montium species subgroup. AB - To investigate the phylogenetic relationships and molecular evolution of alpha amylase (Amy) genes in the Drosophila montium species subgroup, we constructed the phylogenetic tree of the Amy genes from 40 species from the montium subgroup. On our tree the sequences of the auraria, kikkawai, and jambulina complexes formed distinct tight clusters. However, there were a few inconsistencies between the clustering pattern of the sequences and taxonomic classification in the kikkawai and jambulina complexes. Sequences of species from other complexes (bocqueti, bakoue, nikananu, and serrata) often did not cluster with their respective taxonomic groups. This suggests that relationships among the Amy genes may be different from those among species due to their particular evolution. Alternatively, the current taxonomy of the investigated species is unreliable. Two types of divergent paralogous Amy genes, the so-called Amy1- and Amy3-type genes, previously identified in the D. kikkawai complex, were common in the montium subgroup, suggesting that the duplication event from which these genes originate is as ancient as the subgroup or it could even predate its differentiation. Thc Amy1-type genes were closer to the Amy genes of D. melanogaster and D. pseudoobscura than to the Amy3-type genes. In the Amy1-type genes, the loss of the ancestral intron occurred independently in the auraria complex and in several Afrotropical species. The GC content at synonymous third codon positions (GC3s) of the Amy1-type genes was higher than that of the Amy3 type genes. Furthermore, the Amy1-type genes had more biased codon usage than the Amy3-type genes. The correlations between GC3s and GC content in the introns (GCi) differed between these two Amy-type genes. These findings suggest that the evolutionary forces that have affected silent sites of the two Amy-type genes in the montium species subgroup may differ. PMID- 12574859 TI - Phylogeny of a growth hormone-like cytokine superfamily based upon 3D structure. AB - Cytokines are of central importance in the regulation of hematopoiesis, immunity, inflammation, tissue remodeling, and embryonic development. Cytokine research is expected to provide the key to pharmacological manipulation of the immune response and commands the attention of a massive and highly focused biotechnology industry. Based upon the hypothetical secondary and tertiary structures, a superfamily of growth hormone (GH)-like cytokine was identified previously. Here, we report the phylogeny of this superfamily based upon 3D structural data from the Protein Data Bank. First, a retrieving program is designed to abstract their secondary structures and associated atomic coordinates. Helices, digitized as vectors in the Cartesian coordinate system, are collected from the retrieved atomic coordinates at the alpha carbons of the protein backbone. Then the scalar value and vector angle against the reference vector, usually the first vector, are calculated. Furthermore, cluster analysis among various cytokines is performed on their helical scales and helical angles. As a result, GH is close to the cluster formed by ciliary neurotrophic factor and granulocyte colony stimulating factor (CSF); leptin and erythropoietin are in descending order close to the cluster formed by interleukin (IL)-6 and IL-10; the former seven members in the two subgroups above join together and form one group with leukemia inhibitory factor; granulocyte-macrophage CSF, IL-2, IL-4, and IL-5 are in descending order close to the cluster formed by IL-3 and macrophage CSF; and the latter six members form another group. Finally, it is demonstrated that the phylogeny of GH-like cytokines above is consistent with the evolutionary relationship of their gene organization, gene localization, receptor module composition, and receptor module compatibility. PMID- 12574860 TI - Deep origin of plastid/parasite ATP/ADP translocases. AB - Membrane proteins that transport ATP and ADP have been identified in mitochondria, plastids, and obligate intracellular parasites. The mitochondrial ATP/ADP transporters are derived from a broad-specificity transport family of eukaryotic origin, whereas the origin of the plastid/parasite ATP/ADP translocase is more elusive. Here we present the sequences of five genes coding for ATP/ADP translocases from four species of Rickettsia. The results are consistent with an early duplication and divergence of the five ATP/ADP translocases within the rickettsial lineage. A comparison of the phylogenetic depths of the mitochondrial and the plastid/parasite ATP/ADP translocases indicates a deep origin for both transporters. The results provide no evidence for a recent acquisition of the ATP/ADP transporters in Rickettsia via horizontal gene transfer, as previously suggested. A possible function of the two types of ATP/ADP translocases was to allow switches between glycolysis and aerobic respiration in the early eukaryotic cell and its endosymbiont. PMID- 12574861 TI - Cost-minimization of amino acid usage. AB - The negative correlation between the frequencies of usage of amino acids and their biosynthetic cost suggests that organisms minimize costs of protein biosynthesis. Empirical results support that: (1) free-living organisms (Archaea, Bacteria, and Eucaryota) minimize the usage of heavy amino acids more than intracellular organisms (viruses, chloroplasts, and mitochondria), a result confirmed by comparing intracellular Bacteria with other Bacteria; (2) avoidance of amino acids with low impact on protein structure (Chou-Fasman indices) is greater than for those with equal molecular weight but greater structural impact: constraints on protein function limit cost-minimization; (3) amino acid weight minimization (WM) for a protein correlates positively with the protein's expression level and with its size; (4) preliminary results suggest that for different proteins, the evolutionary rate of amino acid replacements correlates negatively with WM in these proteins; (5) results suggest that WM decreases with genome-size; and (6) developmental rates correlate positively with WM (within primates and rodents), even after confounding factors were accounted for. Effects of biosynthetic cost-minimization at whole-organism levels vary with metabolic and ecological strategies. Biosynthetic cost-minimization is an adaptive hypothesis that yields a semi-mechanistic explanation for small differences in allele fitness. PMID- 12574862 TI - Can an arbitrary sequence evolve towards acquiring a biological function? AB - To explore the possibility that an arbitrary sequence can evolve towards acquiring functional role when fused with other pre-existing protein modules, we replaced the D2 domain of the fd-tet phage genome with the soluble random polypeptide RP3-42. The replacement yielded an fd-RP defective phage that is six order magnitude lower infectivity than the wild-type fd-tet phage. The evolvability of RP3-42 was investigated through iterative mutation and selection. Each generation consists of a maximum of ten arbitrarily chosen clones, whereby the clone with highest infectivity was selected to be the parent clone of the generation that followed. The experimental evolution attested that, from an initial single random sequence, there will be selectable variation in a property of interest and that the property in question was able to improve over several generations. fd-7, the clone with highest infectivity at the end of the experimental evolution, showed a 240-fold increase in infectivity as compared to its origin, fd-RP. Analysis by phage ELISA using anti-M13 antibody and anti-T7 antibody revealed that about 37-fold increase in the infectivity of fd-7 was attributed to the changes in the molecular property of the single polypeptide that replaced the D2 domain of the g3p protein. This study therefore exemplifies the process of a random polypeptide generating a functional role in rejuvenating the infectivity of a defective bacteriophage when fused to some preexisting protein modules, indicating that an arbitrary sequence can evolve toward acquiring a functional role. Overall, this study could herald the conception of new perspective regarding primordial polypeptides in the field of molecular evolution. PMID- 12574863 TI - Evolutionary dynamics of large numts in the human genome: rarity of independent insertions and abundance of post-insertion duplications. AB - We determined the phylogenetic positions of 82 large nuclear pseudogenes of mitochondrial origin (numts) within the human genome. For each numt, two possibilities pertaining to its origin were considered: (1) independent insertion from the mitochondria into the nucleus, or (2) genomic duplication subsequent to the insertion. A significant increase in the rate of numt accumulation is seen after the divergence of Platyrrhini (New World monkeys) from the Catarrhini (Old World monkeys, apes and humans). By using pairwise phylogenetic analyses, we were able to demonstrate that this peak in numt accumulation is mostly the result of duplication of preexisting nuclear numts rather than the result of an increase in mitochondrial-sequence insertion. In fact, only about a third of all the numt repertoire in the human nuclear genome is due to insertions of mitochondrial sequences, the rest originated as duplications of preexisting numts. Hence, we conclude that numt insertion occurs at a much lower rate than previously reported. As expected under the assumption that genomic duplications occur at rates that are uninfluenced by content, older numts were found to be duplicated more times than recently inserted ones. PMID- 12574864 TI - The p32K structural protein of the atadenovirus might have bacterial relatives. AB - The primary structure of a novel adenoviral protein referred to as p32K and found exclusively in members of the proposed new genus Atadenovirus was analyzed. The p32K gene sequence was determined from two bovine and one snake adenovirus types. Altogether five different p32K sequences were examined, two of them were obtained from the Gene Bank. The C-terminal part of the protein is conserved and shares similarity with certain bacterial small acid soluble proteins (SASPs). The sequence similarity seems coupled with functional relatedness, i.e. both protein groups are found in structures where the genome of the "dormant" organism is packaged in tight nucleoprotein complexes. In these complexes the DNA is protected against harmful environmental effects until the new reproductive cycle is started with specific protease cleavage of the packaging proteins. Although there is no experimental clue about the role of the p32K proteins, we hypothesize phylogenetic relationship between the two protein groups based on the sequence similarity and the supposed functional similarity. The alignments of these protein groups shows that the conserved part of the p32Ks probably is the result of the duplication of a shorter sequence similar to the SASPs of the Bacilli. PMID- 12574865 TI - Molecular characterization of the recent intragenomic spread of the murine endogenous retrovirus MuERV-L. AB - The mouse genome has been subjected to two successive amplification bursts of the murine endogenous retrovirus MuERV-L after the Mus/Rattus split. The main objective of this work is to characterize in detail the intragenomic spread giving rise to these two murine bursts using full-length MuERV-L proviruses taken from public databases. Phylogenetic analyses led to the identification of elements putatively amplifying during each one of the two burst. Likelihood-ratio tests were used to confirm that elements supposedly arisen during the first burst have been evolving under lower selective constrains, as expected for older insertions. The data reported here suggested an evolutionary dynamics for MuERV-L amplification characterized by the existence of multiple elements simultaneously active during each one of the bursts while only one or very few closely related proviruses from the first burst gave rise to the second one. Finally, more than one third of the proviruses present 100% identity between the 5' and 3' LTRs, strongly indicating that MuERV-L is currently active within the mouse genome. PMID- 12574866 TI - Interspecies comparison of a gene pair with partially redundant function: the rst and kirre genes in D. virilis and D. melanogaster. AB - The D. melanogaster rst and kirre genes encode two highly related immunoglobulin like cell adhesion molecules that function redundantly during embryonic muscle development. The two genes appear to be derived from a common ancestor by gene duplication. Gene duplications have been proposed to be of major evolutionary significance since duplicated redundant sequences can accumulate mutations without detrimental effects for the organism and leave the duplicated genes free to assume novel functions. To address the issue of conservation of the duplicated sequences and their putative redundancy, as well as to identify putative functional divergence of the paralogs during drosophilid evolution, we performed an interspecies comparison of the rst and kirre genes from D. virilis and D. melanogaster. The D. virilis genome contains orthologues of both rst and kirre and hence the duplication took place before the split of the two lineages and has subsequently been conserved. However, whilst the Rst orthologues show a high degree of sequence similarity, this similarity is lower in Kirre orthologues. Especially the intracellular domains of D. virilis and D. melanogaster Kirre sequences are highly divergent: the D. virilis kirre gene lacks the 3'-most exon present in D. melanogaster, which contains motifs conserved between kirre and rst in D. melanogaster. Hence, while each of the two genes is highly conserved at the level of its exon-intron organization, the selection forces acting on the rst and kirre coding sequences are different. These findings are discussed in the light of general evolutionary mechanisms. PMID- 12574867 TI - When being "most likely" is not enough: examining the performance of three uses of the parametric bootstrap in phylogenetics. AB - I show that three parametric-bootstrap (PB) applications that have been proposed for phylogenetic analysis, can be misleading as currently implemented. First, I show that simulating a topology estimated from preliminary data in order to determine the sequence length that should allow the best tree obtained from more extensive data to be correct with a desired probability, delivers an accurate estimate of this length only in topological situations in which most preliminary trees are expected to be both correct and statistically significant, i.e. when no further analysis would be needed. Otherwise, one obtains strong underestimates of the length or similarly biased values for incorrect trees. Second, I show that PB based topology tests that use as null hypothesis the most likely tree congruent with a pre-specified topological relationship alternative to the unconstrained most likely tree, and simulate this tree for P value estimation, produce excessive type I error (from 50% to 600% and higher) when they are applied to null data generated by star-shaped or dichotomous four-taxon topologies. Simulating the most likely star topology for P value estimation results instead in correct type-I-error production even when the null data are generated by a dichotomous topology. This is a strong indication that the star topology is the correct default null hypothesis for phylogenies. Third, I show that PB-estimated confidence intervals (CIs) for the length of a tree branch are generally accurate, although in some situations they can be strongly over- or under estimated relative to the "true" CI. Attempts to identify a biased CI through a further round of simulations were unsuccessful. Tracing the origin and propagation of parameter estimate error through the CI estimation exercise, showed that the sparseness of site-patterns which are crucial to the estimation of pivotal parameters, can allow homoplasy to bias these estimates and ultimately the PB-based CI estimation. Concluding, I stress that statistical techniques that simulate models estimated from limited data need to be carefully calibrated, and I defend the point that pattern-sparseness assessment will be the next frontier in the statistical analysis of phylogenies, an effort that will require taking advantage of the merits of black-box maximum-likelihood approaches and of insights from intuitive, site-pattern-oriented approaches like parsimony. PMID- 12574868 TI - Broad-scale analysis contradicts the theory that generation time affects molecular evolutionary rates in plants. AB - Several studies of plant taxa have concluded that generation time, including annual/perennial life history, may explain molecular evolutionary rate variation in selectively neutral DNA. Unlike in animals, there is little theoretical basis for why generation-time effects would exist in plants. Furthermore, previous reports fail to establish the generality of a generation-time effect in plants because of the small size of the datasets, a large proportion of which compared very widely divergent taxa differing in many characteristics other than generation time. Using 24 phylogenetically independent species pairs, each containing a species with an annual and a species with a perennial life history, and nine species pairs, each containing a tree species with a short and a long minimum generation time, we found no evidence that generation time is related to molecular evolutionary rate variation of the nuclear 18S ITS1 and ITS2 regions. This analysis strongly contradicts the growing belief that evolutionary rates are affected by generation time in plants. Possible reasons for the absence of generation-time effects are discussed, including an evaluation of the cell division theory. PMID- 12574869 TI - Molecular evolution of vertebrate goose-type lysozyme genes. AB - We have found that mammalian genomes contain two lysozyme g genes. To better understand the function of the lysozyme g genes we have examined the evolution of this small gene family. The lysozyme g gene structure has been largely conserved during vertebrate evolution, except at the 5' end of the gene, which varies in number of exons. The expression pattern of the lysozyme g gene varies between species. The fish lysozyme g sequences, unlike bird and mammalian lysozyme g sequences, do not predict a signal peptide, suggesting that the encoded proteins are not secreted. The fish sequences also do not conserve cysteine residues that generate disulfide bridges in the secreted bird enzymes, supporting the hypothesis that the fish enzymes have an intracellular function. The signal peptide found in bird and mammalian lysozyme g genes may have been acquired as an exon in the ancestor of birds and mammals, or, alternatively, an exon encoding the signal peptide has been lost in fish. Both explanations account for the change in gene structure between fish and tetrapods. The mammalian lysozyme g sequences were found to have evolved at an accelerated rate, and to have not perfectly conserved the known active site catalytic triad of the bird enzymes. This observation suggests that the mammalian enzymes may have altered their biological function, as well. PMID- 12574870 TI - Palangeal quantitative ultrasound, phalangeal morphometric variables, and vertebral fracture discrimination. AB - The aim of this study was to evaluate the association among phalangeal morphometric parameters, amplitude-dependent speed of sound (AD-SOS), ultrasound bone profile index (UBPI), and spinal bone mineral density (BMD) and fracture status. One hundred women (controls, mean age 53 +/- 12 years) and 40 osteoporotic women (mean age 59 +/- 7 years) with atraumatic fractures, diagnosed by spinal radiographs, were investigated. Quantitative ultrasound (QUS) assessment was performed using the DBM Sonic 1200. Morphological properties of the phalanges were measured from a digitized X-ray image of the hand acquired using industrial film. Spinal BMD was assessed by dual X-ray absorptiometry (DXA) and quantitative computed tomography (QCT). An increase in medullary canal width and a decrease in cortical thickness with aging were observed from the morphometric analysis of the hand radiographs. This phenomenon can be attributed mainly to endosteal resorption. QUS measurements at the phalanges were not significantly related to finger thickness (r <0.20, n.s.). They were significantly correlated to medullary canal ratio (r = -0.57, P <0.0001, for AD SOS and r = -0.64, P <0.0001, for UBPI) and to cortical thickness (r = +0.52, P <0.0001 for AD-SOS and r = +0.59, P <0.0001 for UBPI). In the discrimination analysis between nonfractured and atraumatic vertebral fracture subjects we found that cortical thickness at the level of the phalanges were similar to lumbar spine BMD. The age and BMI-adjusted odds ratio ranged from 2.0 to 3.1 for QUS, 4.28 for BMD by QCT, 4.1 for BMD by DXA, and 4.1 for cortical thickness. We conclude from these data that phalangeal QUS is related to cortical thickness, which in turn is influenced by endosteal bone resorption occurring in association with spinal osteoporosis. PMID- 12574871 TI - Effects of exercise training on bone remodeling, insulin-like growth factors, and bone mineral density in postmenopausal women with and without hormone replacement therapy. AB - The purpose of this study was to determine the effects of 12 months of weight bearing and resistance exercise on bone mineral density (BMD) and bone remodeling (bone formation and bone resorption) in 2 groups of postmenopausal women either with or without hormone replacement therapy (HRT). Secondary aims were to characterize the changes in insulin-like growth factors-1 and -2 (IGF-1 and -2) and IGF binding protein 3 (IGFBP3) in response to exercise training. Women who were 3-10 years postmenopausal (aged 40-65 years) were included in the study. Women in the HRT and no HRT groups were randomized into the exercise intervention, resulting in four groups: (1) women not taking HRT, not exercising; (2) those taking HRT, not exercising; (3) those exercising, not taking HRT; and (4) women exercising, taking HRT. The number of subjects per group after 1 year was 27, 21, 25, and 17, respectively. HRT increased BMD at most sites whereas the combination of exercise and HRT produced increases in BMD greater than either treatment alone. Exercise training alone resulted in modest site-specific increases in BMD. Bone remodeling was suppressed in the groups taking HRT regardless of exercise status. The bone remodeling response to exercise training in women not taking HRT was not significantly different from those not exercising. However, the direction of change suggests an elevation in bone remodeling in response to exercise training, a phenomenon usually associated with bone loss. No training-induced differences in IGF-1, IGF-2, IGF-l:IGF-2 (IGF-1 : IGF-2), and IGFBP3 were detected. PMID- 12574872 TI - Efferent loop small intestinal vitamin D receptor concentration and bone mineral density after Billroth II (Polya) gastrectomy in humans. AB - Animal studies have demonstrated that the highest concentration of vitamin D receptors (and greatest capacity for active calcium absorption) occurs in the proximal duodenum. By passing the duodenum following Polya/Billroth II gastrectomy could result in the development of a metabolic bone disease and low bone mineral density (BMD). We thus compared the vitamin D receptor (VDR) concentration in mucosal biopsies taken at endoscopy from two functionally corresponding areas of the small intestine: the jejunum (or efferent loop) in 21 patients with a history of Polya/Billroth II gastrectomy and the second part of the duodenum in age/sex-matched control subjects. We also measured the BMD by dual energy X-ray absorptiometry. The mean VDR concentration was not significantly different between the two groups (patients vs controls, fmol/mg protein, mean +/- SE: 34.99 +/- 2.57 vs 34.67 +/- 3.71; P = 0.22), even when subgrouped as males (36.22 +/- 3.16 vs 31.2 +/- 4.24; P = 0.351) or females (31.93 +/- 4.7 vs 43 +/- 6.76; P = 0.193). In Polya/Billroth II gastrectomy patients, the VDR concentration in the efferent loop declined with age (r = 0.78, P = 0.02). In the same group, BMD, as compared with matched controls, was significantly reduced at the lumbar spine (Z-score: patients vs controls: -1.138 vs 0.099, P = 0.01), but not at the femoral neck (Z-score: -0.69 vs 0.7, P = 0.084). There was no correlation between VDR and time since operation or BMD. These results suggest that following Polya/Billroth II gastrectomy, the functional capacity of the jejunal efferent loop in reference to VDR concentration is similar to that of the second part of the duodenum in normal subjects. Therefore, the reduced BMD in our patients, also a common finding in other studies, may not be secondary to the reduced capacity of the VDR system that facilitates the active calcium transport pathway in the proximal small intestine. PMID- 12574873 TI - Urinary cross-linked N-telopeptides of type I collagen levels in patients with rheumatoid arthritis. AB - Osteoclastic activation rather than suppression of bone formation has been suggested to be the dominant process leading to bone loss in rheumatoid arthritis (RA). Although many studies have already shown the correlation of urinary pyridinoline (PYD) and deoxypyridinoline (DPD) levels with RA-related bone loss, urinary cross-linked N-telopeptides of type I collagen (NTx), a more specific marker of bone-derived type I collagen fragments in urine than urinary PYD and DPD in RA, has not been adequately studied. The purpose of the present study was to determine clinical factors that are associated with an increase in urinary NTx levels in patients with RA. One hundred and eighty-four patients with RA and 185 sex- and age-matched controls were enrolled in the study: 71 men, 37-68 years of age (RA: 31, controls: 40); 129 premenopausal women, 30-48 years of age (RA: 67, controls: 62), and 169 postmenopausal women, 48-69 years of age (RA: 86, controls: 83). The correlations of urinary NTx levels, measured by enzyme-linked immunosorbent assay with anatomic grade in the wrist, functional class, duration of disease, steroid use, modified health assessment questionnaire (HAQ) score for the upper and lower extremities, the levels of serum c-reactive protein and rheumatoid factor (RF), erythrocyte sedimentation rate, and/or years since menopause were examined by multiple regression analysis. Urinary NTx levels (nmol BCE/mmol Cr) did not differ significantly between men with RA and controls (53.2 +/- 29.6 vs 41.0 +/- 19.6, respectively), whereas urinary NTx levels were significantly higher in pre- and postmenopausal women with RA than in respective controls (premenopausal women: 57.1 +/- 36.6 vs 42.3 +/- 21.3, P <0.01; women: 76.2 +/- 27.3 vs 57.1 +/- 28.3, P <0.001). In men with RA, no clinical factors were significantly correlated with urinary NTx levels. In premenopausal women with RA, functional class, HAQ score for the upper extremities, and RF were significantly correlated with urinary NTx levels (all P <0.05); in postmenopausal women with RA, functional class and RF were significantly correlated with urinary NTx levels (both P <0.05). These findings suggest that urinary NTx levels were significantly higher only in women with RA than in age-matched controls, and a RA related increase in urinary NTx levels may be associated with physical inactivity and disease activity. PMID- 12574874 TI - Fourier transform infrared imaging spectroscopy (FT-IRIS) of mineralization in bisphosphonate-treated oim/oim mice. AB - Fourier transform infrared microscopy (FT-IRM) and imaging spectroscopy (FT-IRIS) are increasingly used to analyze the molecular components of mineralized tissues. A primary advantage of these techniques is the capability to simultaneously image the quantity and quality of multiple components in histological sections at 7 microm spatial resolution. In the current study, FT-IRM and FT-IRIS were used to characterize bone mineralization in a mouse model of osteogenesis imperfecta (OI) after treatment with the bisphosphonate alendronate (ALN). This application is currently relevant since recent studies have demonstrated great promise for the treatment of children with OI with bisphosphonates, but have not identified bisphosphonate-associated bone quality changes. Growing oim/oim mice, a model of moderate-to-severe OI, were treated with ALN (73 microg ALN/kg/day for 4 weeks followed by 26 mg/kg/day for 4 weeks) or saline from 6 to 14 weeks of age, and mineralization was evaluated in femoral cortical and metaphyseal bone. Infrared vibrations of the mineral (a carbonated apatite) and the matrix phases were monitored. The relative amounts of mineral and matrix present (min:matrix), the relative amount of carbonate present in the mineral (carb:min), and the crystallinity of the mineral phase were calculated. In untreated oim/oim bone, the min:matrix was greater and the crystallinity was reduced (indicative of less mature mineral) in the primary versus the secondary spongiosa, most likely due to the presence of calcified cartilage. With ALN treatment, the oim/oim mm:matrix increased in the secondary spongiosa, but the mineral crystallinity was not changed. In the cortical bone, no changes were evident with ALN treatment. These data demonstrate that in this mouse model, ALN treatment results in increased metaphyseal bone mineralization, but does not improve mineral maturity. PMID- 12574875 TI - Age dependence of in situ termostability of collagen in human bone. AB - The hypothesis of this study is that the in situ thermostability of collagen in bone changes with age and such changes relate to the structural properties of collagen and/or the interaction between the collagen and mineral phases. To test the hypothesis, the effect of age on the in situ thermostability of collagen in human bone and its correlation with the mineral and collagen phases were investigated. In this study, 30 human cadaveric femurs were collected and divided into three age groups: young adults (20-45 years), middle aged (46-69 years), and elderly (over 70 years of age). The in situ thermostability of collagen was assessed using high performance liquid chromatography (HPLC) in terms of the amount of heat-induced collagen denaturation at eight temperatures between 25 degrees C and 140 degrees C. In addition to the density and weight fraction of the mineral and organic phases, the concentration of collagen crosslinks was measured to assess the structural integrity of collagen. The results of this study indicate that the in situ thermostability of collagen increases with increasing age, and such age-related changes correlate with the following: collagen molecular structure, amount of noncalcified collagen, and the fraction and density of the mineral phase in bone. These results suggest that the age related changes in the in situ thermostability of collagen most likely relate to the collagen structure and its interaction with the mineral phase. In addition, the bone remodeling process may play a role in the age-related changes in collagen thermostability because noncalcified collagen is commonly associated with this process. PMID- 12574877 TI - Long-term zoledronic acid treatment increases bone structure and mechanical strength of long bones of ovariectomized adult rats. AB - Zoledronic acid (ZOL) is a highly potent heterocyclic bisphosphonate which has been shown to inhibit bone resorption in short-term experiments in young growing animals. In this investigation we have evaluated the effects of a 1-year administration to mature, ovariectomized (OVX) rats as a model for postmenopausal osteoporosis in order to elucidate (1) the temporal changes in urinary biochemical markers of bone turnover and femoral bone mineral density (BMD), (2) to measure changes of static and dynamic histomorphometric parameters and mechanical strength, and (3) to assess the preventive effects of chronic treatment with ZOL on these parameters. In urine, deoxypyridinoline increased after OVX and was significantly reduced by ZOL administration, indicative of a reduced bone collagen turnover. These changes were accompanied by alterations of tibial cancellous bone: trabecular bone volume and parameters of bone architecture were significantly augmented by ZOL and bone formation rates fell as a consequence of suppressed bone turnover, but were still measurable. No signs of "frozen bone" or osteomalacia could be detected. BMD of the whole femurs rose in sham-operated control animals (SHAM) during the entire experimental period, whereas in OVX animals, BMD plateaued after 32 weeks at a lower level. ZOL at a low dose (0.3 mg/kg/week s.c.) did not alter whole femur BMD, but at higher doses (1.5 and 7.5 mg/kg/week s.c.) BMD increased to the level of the SHAM group. A distinct pattern was noted for the distal quarter of the femur, a region rich in cancellous bone: BMD initially increased in all treatment groups except the OVX group, and at a later stage fell again at a comparable rate irrespective of treatment. Mechanical stability, as assessed by a 3-point bending test, was significantly increased by all doses of ZOL and exceeded OVX and sham-operated controls. The effects on mechanical properties were observed at a low dose which did not measurably increase femoral BMD after 1-year treatment. Multiregression analysis revealed a significant positive correlation between maximum load and BMD, and a significant negative correlation of maximum load with labeled perimeter, a marker of bone formation and turnover. No significant correlation was found with urinary deoxypyridinoline, a marker of bone resorption. The data show that mechanical testing detects improvements of functional bone quality following low dose bisphosphonate treatment which are not identified by standard DXA measurements of BMD. PMID- 12574879 TI - [Spectrum of pathogens and resistance to antibiotics in urinary tract infections and the consequences for antibiotic treatment: study of urology inpatients with urinary tract infections (1994-2001)]. AB - From 1994 to 2001 all uropathogens of urology inpatients were identified and the sensitivity to 14 antibiotics was tested [trimethoprim (TMP)/sulfamethoxazole (SMZ), ciprofloxacin, ampicillin, mezlocillin, ampicillin/sulbactam, piperacillin/tazobactam, cefuroxime, cefpodoxime, cefotaxime, ceftazidime, gentamicin, penicillin, oxacillin, and vancomycin]. The following results were obtained: 1. No general trend toward an increase in resistance was noted during the observation period except for E. coli resistance to TMP/SMZ (25.1% in 2000) and ciprofloxacin (10.4% in 2000). 2. Vancomycin-intermediary staphylococci or vancomycin-resistant enterococci played no role. 3. The lowest resistance to all pathogens was found for piperacillin/tazobactam (8.4% in 2001); carbapenems were not tested. 4. If uropathogens are stratified into gram-positive and gram negative bacteria, for oral administration, ciprofloxacin is the antibiotic with the lowest resistance rate for urinary tract infections with gram-negative pathogens and ampicillin/sulbactam for gram-positive pathogens. 5. Subsequent to further differentiation of the pathogens with simple tests that can be performed after overnight incubation of the culture, empirical antibiotic therapy can then be effectively employed. To draw the correct conclusions from these data, the urologist must either be personally involved in the analytical procedure or receive the interim results promptly. PMID- 12574880 TI - [The disturbing development of resistance in urinary tract infections]. AB - Despite recent caveats about the rapid development of resistance to fluoroquinolones in the treatment of urinary tract infections (UTI), the good tolerability, bioavailability and the broad antibiotic spectrum of fluoroquinolons explain their increased use. We investigated the changes of bacterial spectra and cross resistance profiles in ambulatory and hospitalized UTI patients. A total of 430 positive urine cultures and resistograms were classified according to patient status as either ambulatory or hospitalized and retrospectively analyzed. Cross-resistance profiles of the most effective antibiotics (cotrimoxazol, levofloxacin, amoxicillin/clavulanic acid) were made before an analysis of cost-effectiveness was performed. Whereas Escherichia coli remains the predominant cause of UTI in ambulatory patients, Enterococcus faecalis is the most frequently detected bacterium in the urine cultures of hospitalized patients. This is one reason for the unacceptably high rate of primary resistance of UTI bacteria against cephalosporins. Primary resistance to cotrimoxazol, amoxicillin/cavulanic acid and levofloxacin are impressive and tend to favor the use of levofloxacin. However, high cross-resistance rates reduce the usability of one antibiotic in case of the lack of effectiveness of the other. The broad use of potent antibiotics in hospitals has led to a higher primary resistance and cross-resistance of UTI bacteria in hospitalized patients than in ambulatory patients. The primary resistance of UTI causing bacteria is generally high and worrying. The new fluoroquinolone levofloxacin exhibits surprisingly high primary resistance rates and shares high cross-resistance with other antibiotics that are as effective but much cheaper. Thus, we consider that it should not be a first line treatment option for ambulatory UTI patients in the absence of any resistogram, in order to ensure cost-effectiveness and a slow down in the rapid development of resistance. PMID- 12574881 TI - [Perioperative antibiotic prophylaxis in transperitoneal tumor nephrectomy: does it lower the rate of clinically significant postoperative infections?]. AB - This monocentric prospective randomized study was designed to determine the efficacy of single-shot perioperative antibiotic prophylaxis with 1 g ceftriaxone i.v. in transperitoneal tumor nephrectomy. Eighty-three patients were randomized either into a prophylaxis or a control group: 39 patients received 1 g ceftriaxone i.v. 30 min preoperatively and 44 no study medication. Characteristics of the two groups showed no statistical differences. Postoperative overall infection rates were 7.7% and 27.3% (p=0.007), respectively. Postoperative assessment revealed overall 0 (0%)/7 (15.9%) wound infections, 0 (0%)/2 (4.5%) deep wound infections, 1 (2.6%)/2 (4.5%) pneumoniae, and 2 (5.2%)/3 (6.8%) significant urinary tract infections. In 4 (10.3%)/4 (9.1%) patients, postoperative antibiosis was started without detection of an infectious focus. Overall antibiotic treatment was carried out in 7 (17.9%)/12 (27.3%) patients postoperatively. Costs of antibiotic prophylaxis and/or treatment resulted in 23.60/30.10ZZZ;EUR per patient. Perioperative prophylaxis with 1 g ceftriaxone i.v. decreases postoperative infection rates. Although not all infections have to be treated with antibiotics, there are pharmacoeconomic advantages of such prophylaxis. PMID- 12574882 TI - [Evaluation of symptoms in men with chronic pelvic pain]. AB - According to the National Institutes of Health classification system, chronic non bacterial prostatitis/chronic pelvic pain syndrome (CPPS) is subdivided into an inflammatory (category IIIa) and a non-inflammatory (category IIIb) form. The difference is based on the presence or absence of white blood cells in expressed prostatic secretions, urine after prostatic massage, or semen. This is the only criterion which allows a differentiation between the IIIa and IIIb forms. The symptoms, i.e. pain and urinary complaints of various degrees, are thought to be similar in both forms. These symptoms can be assessed with the Chronic Prostatitis Symptom Index (CPSI) and the International Prostate Symptom Score (IPSS), which are both available in a validated German translation. One hundred and six patients with CPPS were evaluated with CPSI and IPSS. Urinary symptoms troubled all patients. Men with category IIIa had significantly more urinary symptoms when compared to men with category IIIb. There was no difference between the two groups regarding pain and impact on the quality of life. Although pain is thought to be the hallmark of CPPS, the contribution of urinary troubles to the symptoms must not be underestimated. PMID- 12574883 TI - [Hygenic measures for Creutzfeldt-Jakob disease]. AB - The Creutzfeldt-Jakob disease (CJD) belongs to the so-called prion diseases or transmissible spongiform encephalopathies. The iatrogenic transmission of the pathogen by direct contact with infectious tissue, tissue extracts and surgical instruments has been demonstrated. CJD is not an infectious disease in the usual sense. Transmission is not possible by normal social contact nor by nursing measures just as CJD can not be transferred by normal skin contact or contact with blood, serum or other patient' secretions. Therefore, CJD patients do not have to be isolated, because strict conformity to the generally recognized rules of hygiene is completely sufficient. The high resistance of prions to environmental influences requires the combination of different disinfection and sterilization measures before inactivation of the pathogen occurs and it is possible to exclude transmission by medical products. PMID- 12574884 TI - [Infection-induced urinary stones]. AB - Infection stones make up approximately 15% of urinary stone diseases and are thus an important group. These stones are composed of struvite and/or carbonate apatite. The basic precondition for the formation of infection stones is a urease positive urinary tract infection. Urease is necessary to split urea into ammonia and CO(2). As a result, ammonia ions can form and at the same time alkaline urine develops, both being preconditions for the formation of struvite and carbonate apatite crystals. When these crystals are deposited infection stones form. Pathogenetically, various risk factors play a role: urinary obstruction, neurogenic bladder, dRTA, and MSK. If these infections are not treated and the stones are not removed, the kidney will be damaged. Modern methods are available for stone removal, e.g., ESWL and/or instrumental urinary stone removal. Here, especially less invasive methods are preferable. Any treatment must be adjusted to the patient individually. Patients should be examined frequently for recurrent urinary tract infections and stone recurrences, and new infections must be resolutely treated. Good therapy and prophylaxis are possible with present-day treatment modalities. PMID- 12574885 TI - [Gemcitabine in advanced bladder cancer]. AB - MVAC (methotrexate, vinblastine, doxorubicin, cisplatin) has been the standard treatment for patients with advanced urothelial cancer for more than 15 years. Combination chemotherapy including gemcitabine/cisplatin showed similar tumor response and survival rates with a more tolerable toxicity profile in a recent multinational phase III study when compared to MVAC. Effectiveness of gemcitabine as a single agent or in combination with other cytotoxic agents had been investigated before in several phase II studies treating patients with advanced urothelial cancers. The tumor response rate for single agent gemcitabine in advanced urothelial cancers is between 11% and 28%. Tumor response rates rise to 50% when combining gemcitabine with cisplatin, and median survival times between 12 and 15 months can be expected. Triplet therapy schedules including gemcitabine may yield response rates in up to 80% of patients, particularly when used sequentially with other regimens. Further improvement of tolerability during systemic gemcitabine/cisplatin combination therapy without compromising effectiveness was recently demonstrated by a German phase II study when the 4 week schedule was reduced to a 3-week schedule with gemcitabine given on days 1 and 8. PMID- 12574887 TI - [Hospital hygiene is important]. PMID- 12574886 TI - [Antibiotic induced diarrhea and pseudomembranous colitis]. AB - The spore-forming anaerobic bacterium Clostridium difficile has become a serious enteropathogen. Oral and parenteral administration of antibiotics can cause ecological disturbances in the normal intestinal microflora. Suppression of the normal microflora may lead to reduced colonization resistance with subsequent overgrowth by pre-existing, naturally resistant microorganisms, such as C. difficile. C. difficile infection shows a range of clinical presentations between an asymptomatic carrier state, light diarrhea without inflammatory changes, and pseudomembranous colitis. C. difficile infection is acquired by the fecal-oral or environmental-oral routes. From March 2000 through March 2001 we assessed 48 cases of nosocomial antibiotic-associated diarrhea (AAD). Of these, 21 were due to C. difficile (CDAD). Cephalosporin was the agent most commonly associated with CDAD. Avoidance of cephalosporins, strict use of "single shot" prophylaxis, isolation of infected, symptomatic patients in single-bed rooms, improved hygiene and complete room disinfection lead to a rapid decrease of CDAD. The etiology, prognosis and prophylaxis are discussed in this paper. PMID- 12574888 TI - [Urinoma after partial kidney resection]. PMID- 12574889 TI - [Traditional Chinese medicine and phytotherapy]. PMID- 12574890 TI - Molecular mechanisms of inherited ventricular arrhythmias. AB - BACKGROUND: Inherited ventricular arrhythmias such as the long QT syndrome (LQTS), Brugada syndrome, catecholaminergic polymorphic ventricular tachycardia (CPVT), idiopathic ventricular fibrillation (VF), and arrhythmogenic right ventricular cardiomyopathy (ARVC) account for a relevant proportion of sudden cardiac death cases in young patients cohorts. The detailed pathogenetic mechanisms of inherited ventricular arrhythmias are still poorly understood because systematic investigations are difficult to perform due to low patient numbers and the lack of appropriate experimental models. However, recent advances in research and science have identified a genetic background for many of these diseases. PRESENT KNOWLEDGE: In LQTS, various mutations in different genes encoding for cardiac potassium and sodium channel proteins have been identified ("channelopathy"), and initial progress in genotype-phenotype correlation is made. Mutations in the cardiac sodium channel gene have also been identified in a subset of patients with Brugada syndrome, whereas a genetic background has not yet been demonstrated in idiopathic VF and right ventricular outflow-tract tachycardia (RVO-VT). Very recently, mutations in the cardiac ryanodine receptor gene have been identified in CPVT and in a subgroup of patients with ARVC. Although several chromosomal loci were suggested, no other responsible genes or mutations have been found in autosomal dominant forms of ARVC. However, in Naxos disease, a recessive form of ARVC with coexpression of palmoplantar keratoderma and woolly hair, a mutation in the plakoglobin gene has recently been discovered, thus underscoring the potential role of genetic alterations in cytoskeletal proteins in ARVC. FUTURE PERSPECTIVES: In the next years, significant progress in the genetic diagnosis pathophysiologic understanding of disease mechanisms, genotype-phenotype correlation, and the development of gene- or target-directed treatment strategies can be expected in the field of inherited ventricular arrhythmias. CONCLUSION: This review summarizes the current knowledge of the molecular mechanisms, including aspects of pathoanatomy, autonomic innervation, genetics, and genotype-phenotype correlations with their potential implications for diagnosis and treatment of inherited ventricular arrhythmias. PMID- 12574891 TI - [Arrhythmias during remodeling after myocardial infarction]. AB - BACKGROUND: The effect of mechanical on electrical remodeling or electrical instability of the heart shows that it is essential for the prevention of sudden death to avoid or delay mechanical remodeling and neurohumoral activation after myocardial infarction. In other words, patients after myocardial infarction prone to neurohumoral activation need to be treated with ACE inhibitors or perhaps AT1 receptor blockers and beta blockers to maintain electrical stability. ICD INDICATION: MADIT I and MUSTT study showed that patients with severe ventricular dysfunction after myocardial infarction are at high risk of sudden death, especially in presence of electrical instabilities indicated by ventricular arrhythmias. These patients certainly need an automatic implantable cardioverter defibrillator (ICD). It is not clear so far whether or not the indication needs to be extended according to the MADIT II study. In other words, need all postmyocardial infarction patients with reduced pump function an ICD? There is no doubt that many patients with an ejection fraction below 30% have ventricular arrhythmias and fulfil therefore the inclusion criteria for the MADIT I or MUSTT study. In MADIT I, a run of three ventricular premature beats force was sufficient to fulfil the inclusion criteria. CONCLUSION: Another important consequence of the temporal correlation between mechanical and electrical remodeling is that specific attention must be directed to these interrelations in patients after myocardial infarction. Patients who die of sudden death show in comparison to surviving patients a substantial dilatation of the left ventricular during 6 months of observation which parallel the increasing incidence of ventricular premature beats. The consequence for therapy would be that in patients who present with left ventricular dilatation during 6 months after myocardial infarction, electrical instability is present and a high risk of sudden death exists. These patients probably will benefit from an ICD. PMID- 12574892 TI - Sudden cardiac death in dilated cardiomyopathy -- therapeutic options. AB - BACKGROUND: Despite routine use of angiotensin-converting enzyme (ACE) inhibitors, beta-blockers and spironolactone in patients with heart failure due to dilated cardiomyopathy (DCM), these patients still have a considerable annual mortality rate of 5-10%. Sudden unexpected death accounts for up to 50% of all deaths and is most often due to rapid ventricular tachycardia or ventricular fibrillation and less often due to bradyarrhythmias or asystole. THERAPEUTIC OPTIONS: The use of beta-blockers in patients with heart failure has been shown to improve overall mortality considerably. This survival benefit has been demonstrated for bisoprolol, metoprolol and carvedilol. Therefore, one of these three beta-blocking agents should be administered routinely starting with low doses in all patients with New York Heart Association (NYHA) class II or III heart failure in addition to ACE inhibitors, unless there is a contraindication to beta-blocker use. In addition, NYHA class IV heart failure patients have been shown to benefit from carvedilol therapy, if tolerated. The conflicting results of GESICA and CHF-STAT studies do not support a strategy of "prophylactic" amiodarone therapy in patients with DCM in order to prevent sudden cardiac death. Despite growing evidence that implantable cardioverter defibrillator (ICD) therapy results in improved overall survival py preventing sudden cardiac death in patients at high risk for serious arrhythmic events, arrhythmia risk stratification with regard to prophylactic ICD implantation remains highly controversial in patients with DCM. CONCLUSION: This review describes potential arrhythmia mechanisms in DCM and summarizes the results of antiarrhythmic drug trials and of prophylactic ICD trials in patients with heart failure as well as our knowledge concerning arrhythmia risk stratification in patients with DCM. PMID- 12574893 TI - [Arteriosclerosis -- a lifelong challenge]. AB - BACKGROUND: Effective prevention of cardiovascular diseases is of utmost importance in Western civilizations as they are on top of mortality statistics. PRIMARY PREVENTION: Physical exercise plays an important role in optimizing the individuals cardiovascular risk profile. However in the absence of scientifically based studies the potential benefit of physical exercise is often being discussed. Recently by several population-based studies it has been proven that physical exercise is inversely related to long-term cardiovascular mortality in both man and women of all age groups, even after adjustment for other risk factors. SECONDARY PREVENTION: Even for secondary prevention in clinical studies and meta-analyses a reduction of all causes and cardiovascular mortality of 20 45% was found after physical exercise. TYPE OF SPORTS: At present it is being discussed controversially which type of sports would be most beneficial. Recent studies have shown that cardiovascular risk reduction is not only the consequence of vigorous exercise, especially endurance training, but can also be achieved by moderate training programs like walking or increased daily activities. CONCLUSION: Current epidemiological data reveal the necessity to perform more physical activity/exercise training, which would best be integrated in a health oriented lifestyle. An early beginning, even in childhood, is important. PMID- 12574895 TI - [Angiographic results of "atypical" chest pain]. AB - BACKGROUND AND AIMS: Coronary angiography permits evaluation of coronary artery morphology and coronary pathology. It represents an accurate method of defining stenotic coronary lesions. Chest pain may be caused by coronary artery disease as well as by other cardiac and noncardiac disorders. However, sensitivity of clinical evaluation and noninvasive diagnostic assessment in detection of coronary artery disease is limited. Noninvasive diagnostic strategies give inconsistent results in about 10-30%. Here coronary angiography is regarded as an accurate method for appropriate diagnosis. Ist sophisticated apparatus, cost, and invasiveness necessitate well-considered application of this procedure. Therefore, it appears important to analyze coronary angiograms in patients with the referral diagnosis of "atypical" chest pain with inconsistent noninvasive testing or impossibility to perform noninvasive assessment. PATIENTS AND METHODS: We analyzed records of 1,000 consecutive patients (625 men, 375 women, mean age 63.1 years), who underwent coronary angiography at our institution from January 5, 1998 to May 5, 1998. RESULTS: 49 patients (17 women, 32 men; mean age 59 years) were referred due to "atypical" chest pain. 21 (42.9%, nine women, twelve men) of these 49 patients had normal coronary arteries at angiography. 21 (42.9%) patients showed coronary artery disease with a diameter stenosis > 50%. In seven (14.2%) patients, coronary sclerosis with a diameter stenosis < 50% could be observed. Only five (29.4%) of the 17 women but 16 of the 32 men (50%) had coronary artery disease with a diameter stenosis > 50% (p < 0.01). CONCLUSIONS: In unselected patients referred for coronary angiography due to "atypical" chest pain and inconsistent noninvasive testing or impossibility to perform noninvasive assessment. 42.9% had coronary artery disease with a diameter stenosis > 50%. Angiographic evaluation of symptomatic patients with "atypical" signs and symptoms and inconsistent noninvasive testing seems to be appropriate. PMID- 12574894 TI - [Statins and cerebrovascular events]. AB - PREVENTION AND RISK REDUCTION: Statins consistently have shown an ability to reduce the incidence of ischemic stroke. Two meta-analyses corroborated prospective studies with statins and estimated a 30% risk reduction in stroke which is comparable to stroke reduction noted with aspirin. Stroke rate was evaluated in patients without known coronary artery disease in the West of Scotland Primary Prevention (WOSCOPS) trial. An 11% reduction in stroke was observed in WOSCOPS during 5-year follow-up that did not reach statistical significance (p = 0.57). Documentation of clinical benefit with reductions in stroke and coronary events and long-term safety is available only from statin trials with stroke defined as a secondary end point. The strongest evidence relates to patients with CHD. In this population, statistically highly significant reductions have been reported for stroke, cardiovascular mortality, nonfatal myocardial infarction, all-cause mortality, and need for cardiovascular procedures. CONCLUSIONS: Clinical benefit of statin use for stroke prevention in patients without coronary artery disease is not yet proven. Furthermore, it is unknown whether and to what extent differences in lipid and so-called nonlipid mechanisms, half-life, solubility, potency, and effects on other risk factors influence the clinical efficacy. Future trials will be needed to clarify this issue, particularly in patients with no history of coronary artery disease. PMID- 12574896 TI - The effect of aortic valve replacement on coronary flow reserve in patients with a normal coronary angiogram. AB - BACKGROUND: In patients with aortic stenosis and a normal coronary angiogram, a coronary flow reserve (CFR) is impaired. The aim of the present study was to examine the effect of aortic valve replacement (AVR) on the CFR after a long-term follow-up. PATIENTS AND METHODS: 30 patients with aortic stenosis and a normal coronary angiogram were enrolled in the study. CFR measurements were made on 21 patients 123 +/- 137 days before and 497 +/- 167 days after AVR. CFR measurements were carried out according to a standard protocol, with a vasodilator stimulus dipyridamole (0.56 mg/kg for 4 min) and peak diastolic velocity measurements at 6 min. RESULTS: Initially, the average peak gradient of aortic stenosis was 89.5 +/ 22.4 mm Hg. After AVR, it decreased to 26.2 +/- 9 mm Hg. Left ventricular mass was significantly lower after AVR: 354.9 +/- 107.9 g versus 223.8 +/- 73.6 g (p < 0.001). The average baseline diastolic velocity measured by pulsed Doppler in the left anterior descending coronary artery amounted to 62.2 +/- 25.5 cm/s before and 40.1 +/- 13.6 cm/s after AVR. The difference was statistically significant (p < 0.01). The average diastolic velocity at maximum stress equaled 117 +/- 42.8 cm/s pre- and 91.5 +/- 34 cm/s postoperatively (p < 0.005). The calculated CFR before AVR amounted to 1.96 +/- 0.5 and increased to 2.37 +/- 0.8 postoperatively. The difference was statistically significant (p < 0.05). CONCLUSION: Prosthetic AVR is of considerable benefit concerning the CFR in patients with a normal coronary angiogram after a long-term follow-up. PMID- 12574897 TI - Role of heart rate variability in the early diagnosis of diabetic autonomic neuropathy in children. AB - BACKGROUND: Diabetic autonomic neuropathy (DAN) is a major complication of diabetes. DAN has been shown to be closely related to glycemic control. To contribute significantly to the morbidity and mortality of the disease, and to be indicative of an increased risk of cardiovascular events. Tests assessing the function of the autonomic nervous system, such as the response of heart rate and blood pressure to maneuvers stimulating the autonomic nervous system, including deep breathing. Valsalva maneuver and standing, allowed to detect signs of DAN in adolescents; however, the sensitivity of such tests in revealing an early impairment of the autonomic nervous system proved low. Several studies found heart rate variability (HRV) to be useful in assessing the dysfunction of the autonomic nervous system in diabetic children and adolescents, but only few HRV parameters were evaluated in most of them. OBJECTIVE: To study cardiac autonomic nervous system in diabetic children, and to investigate whether the duration of diabetes and the degree of metabolic control are determinants for the development of DAN in children. PATIENTS AND METHODS: We analyzed HRV in 50 asymptomatic patients with insulin-dependent diabetes mellitus (IDDM) and 30 healthy children matched for age and sex. RESULTS: Patients with a history of diabetes > 8 years showed significant alterations of the autonomic nervous system (significant reduction of r-MSSD, pNN50, HF and increase in LF/HF). Conversely, only a reduction in pNN50 was found in patients with a disease duration < 8 years. Furthermore, we also observed significant HRV abnormalities in patients with an impaired metabolic control of diabetes. Compared to controls, patients with glycosylated hemoglobin blood levels (HbA(1C)) > 8% showed a significant reduction of r-MSSD, pHH50 and total power spectrum, whereas no HRV abnormalities were detected in patients with an HbA(1C) < 8%. CONCLUSIONS: HRV analysis can detect early subclinical alterations of the autonomic nervous system in asymptomatic patients with IDDM, which seem to consist mainly in a parasympathetic impairment. Autonomic dysfunction is associated both with the duration and an inadequate metabolic control of the disease. PMID- 12574898 TI - Colchicine for the prevention of postpericardiotomy syndrome. AB - BACKGROUND: Postpericardiotomy syndrome (PPS) is a troublesome complication of cardiac surgery, occurring in 10-45% of cases. Accepted modalities of treatment include nonsteroidal anti-inflammatory drugs, corticosteroids, and pericardiectomy in severe cases. The optimal method for prevention of PPS has not been established. Recent trial data have shown that colchicine is efficient in the secondary prevention of recurrent episodes of pericarditis. The aim of the present study was to evaluate the possible benefit of colchicine for the primary prevention of PPS in patients after cardiac surgery. To the best of our knowledge, this is the first study addressing this issue. PATIENTS AND METHODS: A prospective, randomized, double-blind design was used. The initial study group included 163 patients who underwent cardiac surgery in two centers in Israel between October 1997 and September 1998. On the 3rd postoperative day, the patients were randomly assigned to receive colchicine (1.5 mg/day) or placebo for 1 month. All were evaluated monthly for the first 3 postoperative months for development of PPS. RESULTS: 52 of the 163 patients were excluded because of postoperative complications, noncompliance, or gastrointestinal side effects of treatment. Of the 111 patients who completed the study, 47 (42.3%) received colchicine and 64 (57.7%) placebo. There was no statistically significant difference between the groups in clinical or surgical characteristics. PPS was diagnosed in 19 patients (17.1%), 5/47 cases (10.6%) in the colchicine group and 14/64 (21.9%) in the placebo group. The difference showed a trend toward statistical significance (p < 0.135). CONCLUSIONS: Colchicine may be efficacious for the prevention of PPS in patients after cardiac surgery. Further evaluations in larger clinical trials are warranted. PMID- 12574899 TI - Modified bypass procedure and apicoaortic conduit. Management of coronary artery disease, aortic valve stenosis and porcelain aorta. AB - BACKGROUND: In case of severely calcified ascending aorta, modified operative strategies are required in order to avoid manipulations of the aorta and minimize subsequent cerebral vascular accidents. CASE REPORT: A 73-year-old woman, with a coronary two-vessel disease and aortic stenosis was scheduled for coronary artery bypass grafting and aortic valve replacement. Due to severed calcification of the ascending aorta including the transverse arch, neither cannulation, clamping nor incision of the aorta or its replacement was feasible. Therefore bypass operation was performed using a modified approach. After 1 month, implantation of a valved conduit between the left ventricular apex and the descending aorta through a lateral thoracotomy followed. CONCLUSION: Only in few cases the surgical treatment of a coronary artery disease in combination with left ventricular outflow tract obstruction and heavily calcified ascending aorta has been described. Undoubtedly, creation of an apicoaortic connection is today only indicated in the adult population in a small collective with multiple previous operations or porcelain aorta. PMID- 12574900 TI - [Cardiac dysrhythmia and atypical angina symptoms caused by two bronchogenic cysts]. AB - CASE REPORT: A 41-year-old male saw his general practitioner because of progressive atypical angina symptoms and palpitations for the last 7 years. Chest X-ray showed a mediastinal mass. Further investigation by computed tomography (CT), magnetic resonance imaging (MRI) and cardiac catheterization revealed two bronchogenic cysts. After median sternotomy and pericardial incision, two cystic masses were found on top of and dorsal to the right atrium. Following resection, the patient was free of previously experienced problems related to his atypical angina symptoms. Histological investigation showed no signs of malignancy. CONCLUSION: In patients with atypical angina pectoris the rare case of a bronchogenic cyst has to be considered a possible reason for the symptoms. In addition, patients might show atrium-induced dysrhythmia, coughing with purulent sputum, and pain. CT and MRI are absolutely necessary for exclusion of metastases and aneurysms in the mediastinum. PMID- 12574901 TI - [The isolated rabbit heart: comparison between five different modifications]. AB - BACKGROUND: The isolated heart as an experimental model has been firmly established for more than 100 years. MATERIAL AND METHODS: In this study, five modifications are compared: 1. modified Langendorff apparatus (LA) with modified Krebs-Henseleit (KH) solution a) not containing bovine serum albumin (BSA; n = 13) and b) containing BSA (n = 16), 2. LA with KH solution containing BSH and bovine erythrocytes (n = 14), 3. LA with support rabbit (n = 6), and 4. "working heart" preparation with KH solution, BSA and bovine erythrocytes (n = 16). In the latter modification, no balloon was inserted into the left ventricular cavity, i. e., systemic and coronary circuits were not separated from each other. After completion of the preparation and 20-min stabilization, hemodynamic and metabolic data were assessed while the hearts were contracting in the ejecting mode. Thereafter, protocols for different studies were performed that are not presented here. However, the stability of the modifications within their individual protocols is reported. RESULTS: The results suggest that hearts perfused with KH solution are well suited for short protocols. In spite of the additional costs and time, blood perfusion is required for long-lasting protocols or if changes in coronary flow are to be investigated. CONCLUSIONS: The working heart exhibits both the best function and stability at a relatively low experimental expenditure. Yet, it is not suited for studies where perfusion pressure needs to be changed independent of arterial pressure. PMID- 12574902 TI - Pyodermatitis-pyostomatitis vegetans: two cases and a review of the literature. AB - BACKGROUND: Pyodermatitis-pyostomatitis vegetans (PD-PSV) is a rare vegetating, pustular, eosinophilic, mucocutaneous dermatosis characterized by mucocutaneous lesions of the genital, axillary, and oral regions, as well as on the scalp. OBJECTIVE: We report two patients who were diagnosed with PD-PSV. The published cases of this rare disorder are reviewed. RESULTS: The first patient presented with vegetating pustular plaques of the scalp, vulva, and mouth. The second patient initially showed pustules on fingers and then on toes. Vulva and mouth were later involved. There was no gastrointestinal involvement in either case. In both cases, histology revealed eosinophilic spongiosis with eosinophilic microabscesses and pseudoepitheliomatous hyperplasia. Direct immunofluorescence was negative. CONCLUSION: Pyodermatitis-pyostomatitis vegetans is a rare disorder of unknown cause. It bears similarities to pemphigus vegetans but histology, direct immunofluorescence, and clinical history show the differences. Response to treatment is generally rapid. In several reported cases, an inflammatory gastrointestinal disorder is found in association with the mucocutaneous signs. PMID- 12574903 TI - High-level expression of vascular endothelial growth factor and its receptors in an aphthous ulcer. AB - BACKGROUND: Aphthous ulcers are an extremely common disorder of unknown etiology. These ulcers cause significant morbidity through pain and interference with eating. Thalidomide, an angiogenesis inhibitor, is efficacious for the treatment of aphthous ulcers. METHODS: In situ hybridization was performed on an idiopathic aphthous ulcer using probes specific for the angiogenesis factor vascular endothelial growth factor, and its receptors, in order to determine whether these ulcers are highly angiogenic. CONCLUSIONS: Aphthous ulcers are highly angiogenic. Thalidomide may act to heal aphthous ulcers by inhibiting angiogenesis and promoting reepithelialization. Excess angiogenesis may inhibit reepithelialization in certain types of ulcers, and angiogenesis inhibitors may actually promote wound healing if ulcers are caused by excess angiogenesis. PMID- 12574904 TI - Modulation of the contact hypersensitivity response by AE-941 (Neovastat), a novel antiangiogenic agent. AB - BACKGROUND: AE-941 (Neovastat) is an angiogenesis inhibitor noted to have antiinflammatory properties. OBJECTIVE: We tested Neovastat in a contact hypersensitivity (CHS) model to determine the mechanism of action of its antiinflammatory effects. METHODS: Neovastat was orally administered (200 mg/kg/day) during the sensitization and challenge phases of a murine CHS assay and inflammatory responses were measured. Subsequent assays were performed on mice treated with Neovastat or Cortisone (120 mg/kg/day, IP) and differential mRNA expression of several pro- and antiinflammatory cytokines was quantified using RT-PCR. RESULTS: Neovastat decreased inflammation by 39% when administered during sensitization but did not alter the CHS response when given during the challenge phase. Neovastat significantly induced IL-10 expression in skin and skin-draining lymph nodes (49% and 45%, respectively) and decreased IFNgamma expression in the lymph nodes (35%). CONCLUSION: Antiinflammatory effects of Neovastat observed in CHS could be linked to modulation of cytokines early in the sensitization phase. PMID- 12574905 TI - Subacute cutaneous lupus erythematosus induced by nifedipine. AB - BACKGROUND: Subacute cutaneous lupus erythematosus (SCLE) has been reported to be associated with the use of several drugs, including thiazides, terbinafine, and, rarely, calcium channel blockers. OBJECTIVE: A case of SCLE induced by nifedipine is presented. METHODS AND RESULTS: A 48-year-old white woman developed a papulosquamous and annular eruption in sun-exposed areas during the summer. The patient was taking nifedipine for essential hypertension for four years. Serology showed the presence of antinuclear and anti-Ro/SSA as well as antihistone antibodies. Histopathologic and immunopathologic (granular IgM deposits at the dermoepidermal junction) findings confirmed the diagnosis of SCLE. Nifedipine discontinuation led to rapid improvement with almost complete resolution of skin lesions in one month in the absence of active treatment. Reduction of antinuclear, anti-Ro/SSA, and antihistone antibody levels was documented after six months. CONCLUSION: Nifedipine can cause SCLE after a long period of administration. Antihistone antibodies may be associated with drug-induced SCLE. PMID- 12574906 TI - Facial lipoatrophy following systemic lupus erythematosus. AB - BACKGROUND: Acquired partial lipodystrophy is a rare disorder. An association with systemic lupus erythematosus has been reported. In these cases, an immunologic basis is suggested by the presence of C3 nephritic factor and hypocomplementemia. OBJECTIVE: The following report presents the case of a woman who developed a rapid loss of facial subcutaneous fat a few months after complete spontaneous resolution of cutaneous lesions of lupus. CONCLUSION: Absence of C3NeF in this case suggests that other immunological factors may be involved in the pathogenesis. PMID- 12574910 TI - p53 and mdm-2 expression in Rhabdomyosarcoma of childhood and adolescence: clinicopathologic study by the Kiel Pediatric Tumor Registry and the German Cooperative Soft Tissue Sarcoma Study. AB - Rhabdomyosarcomas (RMS) are the most common malignant soft tissue sarcomas in childhood and adolescence. Despite a large number of publications about this heterogeneous group of tumors, little is known about proliferation, p53 and mdm-2 in relation to histological subtype, clinical parameter, and prognosis of patients. We studied 150 cases of RMS treated in the German Cooperative Soft Tissue Sarcoma Study (CWS) by immunohistochemistry on paraffin-embedded tissue, using antibodies against p53, mdm-2, and Ki-67. The results were correlated with histological subtype, mitotic count, and various clinical parameters. Both p53 and mdm-2 were expressed at low levels and did not show differences between embryonal and alveolar RMS. Tumors of patients with metastatic embryonal RMS showed significantly higher levels of p53 protein than nonmetastatic tumors. This might be a clue to an important role of p53 in metastatic embryonal RMS. Nevertheless, neither p53 nor mdm-2 showed any correlation to prognosis. Proliferation measured by Ki-67 immunostaining (KiS5 antibody) or mitotic count did not show significant differences between embryonal and alveolar RMS. In addition, these parameters did not correlate with response to therapy or prognosis. In conclusion, we could not demonstrate that any of the investigated parameters had an influence on prognosis of RMS. p53 protein overexpression might be a crucial step in metastatic disease for patients with embryonal RMS. PMID- 12574911 TI - Adenovirus ascending cholangiohepatitis. AB - Three children, two with liver transplants and one with acquired human immunodeficiency virus (HIV) infection, presented with hepatitis accompanied by elevated gamma glutamyl transpeptidase. Biopsies revealed cholangiohepatitis caused by adenovirus infection. There was a progressive loss of interlobular bile ducts in two of the patients. In one patient, infection of the biliary tree was marked by a necrotizing cholangitis, with adenoviral inclusions noted in the biliary epithelium. In each patient, there was evidence of adenovirus gastrointestinal infection. This is the first report of adenoviral infection of the biliary tree in humans. It is hypothesized that adenovirus cholangiohepatitis occurs as a result of ascending infection from the gastrointestinal tract to the biliary tree. PMID- 12574913 TI - Patterns of congenital lower urinary tract obstructive uropathy. PMID- 12574916 TI - Lymphomatoid granulomatosis: unusual presentation in a pediatric patient. PMID- 12574917 TI - Pediatric pathologists must know left from right: a case of heterotaxy. PMID- 12574918 TI - Pediatric pathology: a case report highlighting its singular niche. PMID- 12574919 TI - Classic adamantinoma with osteofibrous dysplasia-like foci and secondary aneurysmal bone cyst. AB - Adamantinoma, a rare bone lesion of the tibia and fibula, has two distinct variants, classic adamantinoma and osteofibrous dysplasia-like adamantinoma. Composite lesions have not been described. Aneurysmal bone cyst is a benign cystic lesion which may also occur in the tibia and fibula. We report an unusual case of classic adamantinoma with osteofibrous dysplasia-like areas and foci of secondary aneurysmal bone cyst with prominent giant cells. A lesion was diagnosed in a 17-year-old girl with a 14-year history of a slowly enlarging left tibial mass and increasing deformity. Pathologically, the predominant pattern was classic adamantinoma, with minor foci of osteofibrous dysplasia-like adamantinoma and areas of secondary aneurysmal bone cyst with abundant multinucleated giant cells. We report the clinical, radiologic, and pathologic features of this case, and summarize lesions associated with secondary aneurysmal bone cyst. To our knowledge, the association of adamantinoma with secondary aneurysmal bone cyst has not been previously reported. PMID- 12574920 TI - Giant botryoid fibroepithelial polyp of bladder with myofibroblastic stroma and cystitis cystica et glandularis. AB - A 3-year-old boy presented with a single episode of gross hematuria and no history of previous urinary tract disorder. Imaging studies revealed a large complex polypoid filling defect in the bladder lumen. Several attempts at transurethral biopsy and cytological examination of the urine revealed clumps of benign epithelial cells, but suspicion of a malignant neoplasm such as rhabdomyosarcoma remained high and the lesion was resected. The specimen measured 15 cm, had a narrow zone of attachment to the bladder mucosa, and was grossly botryoid. Changes typical of cystitis cystica et glandularis were present at and near all surfaces. Myxoid stroma contained scattered benign fibroblasts, myofibroblasts, and smooth muscle cells. Inexplicably, one of two karyotyped stromal cells demonstrated a translocation usually associated with rhabdomyosarcoma. This child is well without evidence of bladder abnormality 1.5 years after surgery. PMID- 12574922 TI - Laparoscopic treatment of simple hepatic cysts and polycystic liver disease. AB - BACKGROUND: The authors present their experience in the laparoscopic management of hepatic cysts and polycystic liver disease (PLD). METHODS: Between January 1996 and January 2002, 16 patients underwent laparoscopic liver surgery. Indications were solitary giant cysts (n = 10) and PLD (n = 6). Data were collected retrospectively. RESULTS: Laparoscopic fenestration was completed in 15 patients. Median operative time was 80 min. There was no deaths. Complications occurred in four patients: one patient with a solitary liver cyst experienced diarrhea, while a pleural effusion, a bleeding from the trocar-insertion site, and ascites occurred in three patients with PLD. Median follow-up was 34 months. There was one asymptomatic recurrence (11%) in one patient with a solitary cyst. Two patients with PLD had a symptomatic recurrence of a liver cyst. CONCLUSION: Laparoscopic fenestration could be the preferred treatment of solitary liver cysts and PLD. Adequate selection of patients and type of cystic liver together with a meticulous surgical technique are recommended. PMID- 12574923 TI - Complete fundoplication has similar long-term results in patients with and without esophageal body dysmotility. AB - BACKGROUND: Partial fundoplication is advocated for the treatment of gastroesophageal reflux disease in patients with poor esophageal body function. We hypothesized that a complete floppy wrap may be just as safe in patients with poor esophageal motility. METHODS: A retrospective, case-control study was performed on patients who underwent a complete fundoplication and had poor esophageal motility. Study patients were matched with controls with normal esophageal body pressures according to sex, age, and duration of reflux symptoms. Patients were followed up and interviewed using a modified symptom and life quality questionnaire. RESULTS: Twenty-two patients and 22 matched controls underwent a complete fundoplication. The mean esophageal body pressure was 42.1 and 87.5 mmHg in the study and control groups, respectively (p <0.05). Average time to resolution of dysphagia was 10.1 weeks in the study group and 12 weeks in the control group. All patients but 1 (control) graded their life quality improvement as good to excellent. CONCLUSION: Our data suggest that a 360 degrees fundoplication has similar long-term results regardless of esophageal body motility. We suggest that a partial fundoplication may be reserved for patients with severe esophageal body dysfunction. The role of manometry in the preoperative workup should be reassesed: it may be mandatory only in patients with preoperative dysphagia or when achalasia is suspected. PMID- 12574924 TI - Comparison of conventional and gaze-down imaging in laparoscopic task performance. AB - BACKGROUND: In video-assisted laparoscopy, the image is usually displayed on a monitor placed at approximately eye level. Video projection systems project the image onto a screen placed close to the hands. This is said to be ergonomically superior. To evaluate this approach, a proprietary projection system (PS) was compared to a monitor display (MD). METHOD: The resolution, ghosting, flickering, glare, contrast, color smear, and color matching of the two modalities were compared. A bowel-suturing task was employed to evaluate performance differences. RESULTS: The image displayed by the first-generation PS is inferior to that of the MD in contrast and resolution measures, but it is comparable in the other image qualities. No significant differences in task performance were identified. CONCLUSIONS: The first-generation PS does not confer performance or comfort advantages over an MD. The theoretical advantages of the gaze-down stance are likely to be realized only if a high-quality projector is used. PMID- 12574925 TI - Randomized controlled trial of laparoscopic versus open colectomy for advanced colorectal cancer. AB - BACKGROUND: After confirming a favorable outcome of laparoscopic surgery for early colorectal cancer, we conducted a randomized controlled trial to compare short-term outcomes of laparoscopic and open colectomy for advanced colorectal cancer. METHODS: Fifty-nine patients with T2 or T3 colorectal cancer were randomized to undergo laparoscopic (n = 29) or open (n = 30) colectomy. Median follow-up was 20 months (range, 6-34 months). RESULTS: Operative time was longer (p <0.0001) and blood loss (p = 0.0034) and postoperative analgesic requirement were less in the laparoscopic group than in the open group. An earlier return of bowel motility and earlier discharge from the hospital (p = 0.0164) were observed after laparoscopic surgery. Serum C-reactive protein levels on postoperative days 1 (p <0.0001) and 4 (p = 0.0039) were lower in the laparoscopic group than in the open group. Postoperative complications did not differ between the two groups. CONCLUSION: Laparoscopic surgery for advanced colorectal cancer is feasible, with favorable short-term outcome. PMID- 12574926 TI - A comparison of holding strength of various surgical clips. AB - Due to advances in clip design, new types of surgical ligation clips are available that may reduce clip failure and improve function, especially for 5-mm clip appliers. We tested a preformed spring clip based on a 5-mm applier against other commonly used clips from 5- and 10-mm multiloading clip appliers. Five different clips were compared using a digital force gauge to test perpendicular pulloff force after application to tubing of varying thickness and compliance. The average pull-off force for commonly used 5-mm and 10-mm clip appliers was 0.29 +/- 0.06 and 0.35 +/- 0.05 kg, respectively. Average pull-off force for the new preformed spring clip was 0.53 +/- 0.07 kg (p <0.0001). The new clip technology has significantly higher pull-off force than other currently used clips. This may be particularly beneficial in procedures requiring 5-mm clip appliers, which are known to place clips with lower holding force. PMID- 12574927 TI - Successful laparoscopic Ladd's procedure and appendectomy for intestinal malrotation with appendicitis. AB - We successfully performed a laparoscopic Ladd's procedure and an appendectomy in a 15-year-old girl with intestinal malrotation and appendicitis. She had tenderness and rebound pain in the umbilicus and left lower abdominal quadrant. Blood analysis revealed a moderate inflammatory response. Enhanced computerized tomography (CT) scanning revealed a whirl-like pattern and a superior mesentric vein (SMV) rotation sign in the mesentry of the small intestine. A swollen appendix was seen just below the umbilicus. An upper gastrointestinal (GI) radiological series confirmed agenesis of Treitz's arch. The patient was diagnosed as having a nonrotation type of malformation accompanied by acute appendicitis. She underwent a laparoscopic Ladd's procedure, an appendectomy, peritoneal lavage, and drainage. The technique for this procedure and its effectiveness are briefly discussed. PMID- 12574928 TI - Combined use of high-frequency jet ventilation and abdominal lift for laparoscopic cholecystectomy in a patient with glottic impairment. AB - Effective airway management during laparoscopic anesthesia is important to minimize the adverse consequences of the carbon dioxide (CO2) pneumoperitoneum (PP). During PP, reduced respiratory excursion and tidal volumes with increased CO2 absorption may lead to hypoxia, hypercapnia, and respiratory acidosis. Although these problems can usually be avoided by use of positive pressure ventilation and an endotracheal tube, patients with a restricted airway who cannot be intubated pose a unique challenge. High-frequency jet ventilation (HFJV) has been described as an alternative to endotracheal intubation in other settings. The use of the small-diameter jet tube allows relatively unobstructed access to the larynx during laryngeal surgery. In patients with glottic impairment related to vocal fold immobility, jet ventilation allows positive pressure ventilation without the use of an endotracheal tube or tracheostomy in cases where lung and diaphragmatic compliance permit adequate excursion for ventilation and glottal diameter permits an adequate outflow of air. In this report, we describe the successful use of HFJV combined with an abdominal lifting technique and low-pressure PP for laparoscopic surgery in a patient with glottic compromise related to vocal fold immobility. Using these techniques, a laparoscopic cholecystectomy was performed successfully without endotracheal intubation or the need for a tracheostomy. PMID- 12574929 TI - Chemical peritonitis: a rare complication of an iatrogenic ovarian dermoid cyst rupture. AB - Laparoscopy is the standard method for the diagnosis and treatment of ovarian dermoid cysts, which represent 10% to 15% of all ovarian tumors. This procedure offers many advantages including less postoperative pain, lower risk of wound complication, early ambulation, and more rapid convalescence. The risk of adhesion formation also is reduced. However, in the case of an ovarian dermoid cyst, the laparoscopic approach could result in chemical peritonitis caused by the spilled contents of a ruptured dermoid cyst. We report a case of chemical peritonitis after rupture of an ovarian dermoid cyst during laparoscopy. Two operations were required, and medical treatment was not beneficial. We discuss a possible new medication for the treatment of this complication and compare this case with other recent reports. PMID- 12574930 TI - Tracheobronchial amyloidosis treated with rigid bronchoscopy and stenting. AB - Tracheobronchial amyloidosis (TBA) is an uncommon disease that can cause airway obstruction. We present a case of TBA in a 20-year-old man that was treated successfully with rigid bronchoscopy and stenting. The patient presented with progressive dyspnea despite having had a tracheostomy fashioned at another institution. Airway obstruction secondary to TBA was found distal to the tracheostomy. The amyloid protein subtype was AA, which is uncommon and is seldom of clinical significance in the respiratory tract. The patient underwent rigid bronchoscopy to remove the amyloid protein causing the airway obstruction. A Dumon silicone stent was then inserted to alleviate the obstruction. Thereafter, he recovered well and was discharged without a tracheostomy. This report shows that in patients with TBA causing airway obstruction, excellent results can be obtained with rigid bronchoscopy and stenting of the obstructing lesion. PMID- 12574931 TI - Laparoscopic repair of an internal supravesical hernia: a rare hernia causing small bowel obstruction. AB - The application of diagnostic laparoscopy in emergency surgery has facilitated a wide range of endoscopic operative procedures. We report an extremely rare case of a patient who had a bowel obstruction caused by an internal supravesical hernia that was repaired via a minimally invasive technique. Abdominal computed tomography (CT) showed signs of small bowel obstruction: the cause was thought to be an invagination due to a small bowel tumor. Laparoscopic exploration of the dilated small bowel segments allowed the diagnosis of supravesical hernia. Reduction was performed with slight traction, and the hernial orifice was closed with intracorporeal sutures. To our knowledge, this is the first repair of an internal supravesical hernia ever to receive herniorraphy based on laparoscopic techniques. The mean starting time for bowel-function and mean hospital stay following the laparoscopic release of the intestinal obstruction were significantly shorter than is typically seen with standard techniques. PMID- 12574932 TI - Ruptured subcapsular hematoma after laparoscopic cholecystectomy attributed to ketorolac-induced coagulopathy. AB - Ketorolac is the first injectable nonsteroidal antiinflammatory drug used as an analgesic in the perioperative period. However, gastrointestinal bleeding is a risk associated with its perioperative administration. A 23-year-old woman was admitted for elective laparoscopic cholecystectomy. Her medical history was unremarkable except for a complaint of intermittent right upper quadrant pain for several months. The operative procedure was uneventful. Thirty milligrams of ketorolac were given intravenously just prior to termination of surgery. Eighteen hours after surgery, the patient developed right upper quadrant pain associated with tachycardia and hypotension. Abdominal computed tomography (CT) scan demonstrated a large subcapsular hematoma of the liver. A few hours later, the hemodynamic condition worsened, and the patient was taken to the operating room. Laparoscopic exploration showed a ruptured subcapsular hematoma with active bleeding. No evidence of parenchymal injury of the gallbladder bed was found. The hematoma was evacuated and hemostasis was performed laparoscopically. Ketorolac has a strong antiplatelet activity and further acts by the inhibition of platelet function, which may last as long as 24 h after the last administration. Surgeons and anesthesiologists should be aware that ketorolac may cause or aggravate bleeding. PMID- 12574933 TI - Wilkie's syndrome. AB - A superior mesenteric artery syndrome (SMAS) was diagnosed in two young women with, respectively, a 2- and 1-year history of postprandial vomiting and epigastric pain. The patients underwent a laparoscopic duodenojejunal bypass, and resumed a normal diet on the fifth postoperative day. The patients are still symptom-free with patent anastomosis on gastrointestinal radiographic control at 24 and 6 months, respectively, following their operation. Herein we also describe the varying clinical presentation of this rare syndrome, as well as treatment options. We conclude that laparoscopic duodenojejunostomy offers a new therapeutic approach to SMAS. It is reliable and safe; the operating time is acceptable; and diet recovery and hospital stay are both short. However, these preliminary results still need to be confirmed by further observations. PMID- 12574934 TI - Botulinum toxin type A (Dysport) for the treatment of Zenker's diverticulum. AB - The standard treatment for Zenker's diverticulum (ZD) is surgical, but transendoscopic section of the diverticular spur has also been performed with good results. We treated two patients with the injection of 300-350 IU of botulinum toxin type A (Dysport) into the diverticular spur. Both patients were dysphagic because of a large ZD. A few hours after the treatment, the patients were able to return to a normal, nonrestricted diet. They remain symptom-free 12 and 3 months later, respectively. No complications have been observed. As a result of its ease of application and efficacy, this treatment can be recommended for patients who are unable to undergo the surgical operations; it can be considered a valid alternative to the other endoscopic procedures. PMID- 12574935 TI - Laparoscopic repair of Morgagni hernia and paraesophageal hernia on the same patient. AB - Morgagni hernias have been recognized with increasing frequency as a source of abdominal pain and dyspnea in adults. Morgagni hernias are rarely accompanied by paraesophageal hernias. We report a case of Morgagni hernia associated with paraosephageal hernia, both repaired laparoscopically. On the 65-year-old woman patient, diaphragmatic defect and paraesophageal hernia were closed with primary sutures, and Hill-type gastropexy was performed successfully. The procedure lasted 115 min. The patient was discharged 5 days after surgical treatment and there were no complications following the operation. Primary closure with direct sutures is rapid, simple, and effective and can be combined with other laparoscopic procedures, as in our case. It can be performed by surgeons trained in intracorporeal suturing and knotting in all kinds of hospitals. The laparoscopic approach to Morgagni hernias minimizes trauma and shortens postoperative hospital stay, and patients have a comfortable postoperative period. PMID- 12574936 TI - Laparoscopic repair of traumatic rupture of the left diaphragm cupola with prosthetic mesh. AB - We present the case of a 23-year-old patient who had a car accident with a steering wheel trauma 6 months previously. The patient complained of abdominal pain, getting easily tired, and breathing difficulties. A diagnosis of traumatic diaphragmatic hernia was established by a contrast radiographic examination. Laparoscopy confirmed a rupture of the left diaphragmatic cupola and prolapse of the stomach, small intestine, and colon in the left thoracic cavity. Laparoscopic repair of the diaphragm with polypropylene mesh was performed. The patient recovered rapidly and was discharged 72 h after the operation. The control radiographs and computed tomography (CT) contrast investigations showed no recurrence. The patient's preoperative complaints had resolved completely. The case is interesting because traumatic diaphragmatic hernia is sometimes difficult to diagnose. Its laparoscopic treatment is still a challenge for modern surgery, and there are not enough publications on the problem. PMID- 12574937 TI - Acute appendicitis after laparoscopic treatment of acute epiploic appendagitis. AB - Epiploic appendagitis (EA) is a rare cause of right lower quadrant (RLQ) abdominal pain. We report an unusual case of acute gangrenous appendicitis that developed after laparoscopic treatment of an EA. A 62-year-old man underwent laparoscopy for RLQ abdominal pain. EA was found and a resection was performed. The appendix, which was macroscopically normal, was left undisturbed. One week later, the patient was operated on for acute gangrenous appendicitis. Histologic examination separately confirmed both diagnoses. The definitive outcome was uneventful. The exact origin of this unusual case is unknown: Could acute appendicitis have been secondary to laparoscopic manipulation or initially missed? We conclude that acute appendicitis may be either missed or induced by laparoscopy for RLQ abdominal pain. PMID- 12574938 TI - Sizable pseudoaneurysm of the abdominal aorta after laparoscopic cholecystectomy: a report of a serious complication. AB - After the first successful laparoscopic cholecystectomy in Europe by Phillipe Mouret, the method quickly became established all over the world. In the meantime, its advantages and complications have been identified. In rare cases, major vascular injuries occur as serious and deadly complications of laparoscopic surgery. We present the case of a patient with a sizable pseudoaneurysm of the abdominal aorta that was identified 5 months after the performance of a laparoscopic cholecystectomy at another hospital. Thereafter, we review the literature in search of some methods that can be used to avoid vascular injuries. PMID- 12574939 TI - Chromosome 18 replaced by two ring chromosomes of chromosome 18 origin. AB - We here describe the first example of the replacement of an autosome by two ring chromosomes originating from the missing chromosome, presented in a patient with a single chromosome 18 and two additional ring chromosomes. Detailed fluorescence in situ hybridization (FISH) analysis revealed the chromosome 18 origin of both ring chromosomes and characterized the small and the large ring chromosome as derivatives of the short and long arm of chromosome 18, respectively. The loss of subtelomeric regions of the short and the long arm of chromosome 18 in the ring chromosomes was confirmed by FISH studies. Molecular studies showed the exclusive presence of the paternal alleles for microsatellite markers located distal to the short and long arm loci D18S843 and D18S474, respectively. This indicates the maternal origin of both rings and provides evidence for substantial deletions of the distal parts of both arms of chromosome 18 in the ring chromosomes. The dysmorphic features of the patient can be explained by these deletions in both chromosome arms, as the clinical findings partly overlap with observations in 18p and 18q-syndrome and are similar to some cases of ring chromosome 18. Centromere misdivision is suggested as one mechanism involved in the formation of the ring chromosomes. PMID- 12574941 TI - The future of association studies of common cancers. PMID- 12574940 TI - Linkage of autosomal dominant common variable immunodeficiency to chromosome 5p and evidence for locus heterogeneity. AB - Common variable immunodeficiency (CVID, OMIM 240500) and selective immunoglobulin A deficiency (IgAD) are the most frequent primary immunodeficiencies in humans. Of the cases with CVID/IgAD, 20%-25% are familial, but the only previous claims of linkage or association are to the HLA region on chromosome 6p. We report the results of a genome-wide scan in three multiplex families with CVID, IgAD, and dysgammaglobulinemia, where affection is inherited in an autosomal dominant pattern. Two of the families are consistent with linkage to the telomeric region of chromosome 5p, whereas the third is consistent with linkage to the HLA region. Using a locus heterogeneity model and a conservative penetrance model, we obtained a LOD score of 3.35 for the 5p region. We sequenced the exons of one promising candidate gene within this region (PDCD6, also known as ALG-2) but found no causative mutation. PMID- 12574942 TI - Genomic basis of cystathioninuria (MIM 219500) revealed by multiple mutations in cystathionine gamma-lyase (CTH). AB - Hereditary cystathioninuria (MIM 219500) is presumed to be caused by deficiency of the activity of cystathionine gamma-lyase (cystathionase; CTH EC 4.4.1.1), which is normally required for the conversion of methionine into cysteine. To date, no mutations have been described among patients with cystathioninuria. From genomic DNA, we sequenced CTH in four unrelated probands with cystathioninuria. We found two nonsense mutations, namely exon 8 c.940-941delCT and exon 11 c.1220delC, and two missense mutations, namely exon 2 c.356C>T (T67I) and exon 7 c.874C>G (Q240E). All affected subjects were either simple homozygotes or compound heterozygotes. A common non-synonymous single nucleotide polymorphism in exon 12, namely c.1364G>T (S403I), was also identified and characterized in four ethnic groups. The reagents described in this report make the molecular diagnosis of cystathioninuria possible, allowing for studies of phenotype-genotype correlation. Also, the availability of a common non-synonymous SNP can allow for testing of association of the CTH gene with biochemical traits affected by trans sulfuration, such as plasma concentrations of homocysteine or even cystathionine itself, in addition to more downstream clinical phenotypes, such as vascular disease. PMID- 12574943 TI - Brain embolism - Answers to practical questions. AB - Embolism of cardiac origin accounts for about 1/4 of ischemic strokes. Strokes due to cardioembolism are in general severe and prone to early and long-term recurrence. Cardioembolism can be reliably suspected in clinical grounds but is often difficult to document. Magnetic resonance imaging, transcranial Doppler, echocardiogram, Holter and electrophysiological studies can help to demonstrate the source of cardioembolism. Aortic arch atheroma is an emerging embolic source, but non-valvular atrial fibrillation remains the commonest cause of cardioembolic stroke. Despite its preventive potential, continuous oral anticoagulation is prescribed for less than half of the patients with atrial fibrillation with risk factors for embolism and no contraindications for anticoagulation. The embolic risk of patent foramen ovale is low except when combined with an atrial septal aneurysm. Available evidence does not support routine immediate anticoagulation of acute cardioembolic stroke. PMID- 12574944 TI - Diagnosis of hereditary neuropathies in adult patients. AB - This paper reviews the clinical diagnostic approach to hereditary neuropathies in adults by analysing: elements that point to a neuropathy of inherited origin, different modalities of presentation, laboratory and instrumental diagnostic tests, including molecular tests, symptoms and signs of involvement of other organs. Different phenotypes may be identified according to: disease course; involvement of motor, sensory, autonomic fibres; site of lesion (neuropathy versus neuronopathy); calibre of involved fibres (small-fibre versus large-fibre neuropathy); presence of distinctive symptoms (neuropathic pain); involvement of other organs or apparatus. Charcot-Marie-Tooth disease, Familial Amyloid Polyneuropathy, Hereditary Sensory and Autonomic Neuropathy, Fabry disease, Tangier disease, Porphyric Neuropathies, Refsum disease, Hereditary Neuropathy with liability to Pressure Palsies, Hereditary Neuralgic Amyotrophy, and other rare disorders involving the peripheral nervous system are reviewed. PMID- 12574945 TI - Phenotypical variability of expanded alleles in the TATA-binding protein gene. Reduced penetrance in SCA17? AB - Trinucleotide expansions in the gene for the TATA-binding protein (TBP) have recently been described in cerebellar ataxia associated with dementia, pyramidal tract and basal ganglia symptoms. Expansions above 45 repeat units are commonly considered pathological, causing SCA17. Here, we present a German kindred with four siblings affected by cerebellar ataxia, chorea and dementia. Molecular genetic analysis yielded an expanded SCA17 allele coding for 48 glutamine residues that was transmitted from the mother to all of her six children. Apparently, the expanded allele does not cosegregate with the disease phenotype since the mother and two of the siblings do not show any clinical abnormality. This appears to be the first description of non-penetrance in SCA17. PMID- 12574946 TI - A clinical and pathological study of a Japanese case of Amyotrophic Lateral Sclerosis/Parkinsonism-Dementia Complex with family history. AB - This report concerns a Japanese family with neuropathological findings consistent with amyotrophic lateral sclerosis/parkinsonism-dementia complex (ALS/PDC) in the Island of Guam. The proband was a 68-year-old woman with an 8-year history of parkinsonism which was followed by psychiatric symptoms and neurogenic amyotrophy 5 years after the onset. She had a family history of parkinsonism associated with dementia in all of her three siblings. They grew up in the Hobara village, a focus of amyotrophic lateral sclerosis in the Kii Peninsula of Japan in their childhood. Their parents were not consanguineous nor natives of the Kii Peninsula. The brain weight was 1040 g and there were mild frontal lobe atrophy, moderate atrophy of pes hippocampi, decoloration of the substantia nigra and locus coeruleus, and atrophy of the anterior root of the spinal cord. The microscopic examinations revealed degeneration of CA1 portion of the hippocampus to the parahippocampus gyrus, substantia nigra, locus coeruleus and spinal anterior horn with Bunina bodies. The spinal pyramidal tracts also mildly degenerated. Neurofibrillary tangles (NFT) were observed in the cerebral cortex, especially in the cortices from hippocampus to lateral occipitotemporal gyri, basal nucleus of Mynert, basal ganglia, thalamus, substantia nigra and widespread regions of the central nervous system through the brainstem to spinal cord including the nucleus of Onufrowitcz. In spite of a small amount of the senile plaques in the cerebral cortex and Lewy bodies in the substantia nigra and locus coeruleus, abundant NFT were distributed mainly in the third layer of the cerebral cortex, which is the characteristic feature of ALS/PDC. Thus, this was likely to be an ALS/PDC case outside the Guam Island. A tau mutation was not found on DNA analysis. PMID- 12574947 TI - Beta-Interferon treatment does not always slow the progression of axonal injury in multiple sclerosis. AB - Progression of disability in multiple sclerosis (MS) appears related to axonal damage, which is at least in part associated with white matter lesions. Beta interferon (BIFN) substantially reduces new inflammatory activity in MS and a recent report suggested that it may reverse a component of axonal injury. To test the generalisability of this conclusion, particularly in a population with relatively active disease, we used magnetic resonance spectroscopy measures to test whether BIFN can reverse or arrest progression of axonal injury in patients with MS. Eleven patients with a history of active (median, 1.5 relapses/year) relapsing-remitting MS were treated with BIFN and responses to treatment were monitored with serial MRI and single voxel magnetic resonance spectroscopic measurements of relative concentrations of brain N-acetylaspartate (NAA), a measure of axonal integrity from a central, predominantly white matter brain region. BIFN treatment was associated with a significant reduction in relapse rate (p = 0.007) and white matter water T2 relaxation time (p = 0.047) over 12 months. Also consistent with a treatment effect, white matter T2-hyperintense lesion loads did not increase. However, the central white matter NAA/creatine ratio (NAA/Cr, which was reduced over 16 % in patients relative to healthy controls at the start of treatment), continued to decrease in the patients over the period of observation (mean 6.2 % decrease, p = 0.02). For individual patients the magnitude of the NAA/Cr decrease was correlated with the frequency of relapses over the two years prior to treatment (r = -0.76, p = 0.006). These data suggest that reduction of new inflammatory activity with BIFN does not invariably halt progression of axonal injury. Nonetheless, there appears to be a relationship between the rate of progression of axonal injury and relapse rate over the previous two years. The consequences of reduced inflammation on pathological progression relevant to disability therefore may be present, but substantially delayed. Alternatively, distinct mechanisms may contribute to the two processes. PMID- 12574948 TI - Bilateral subthalamic stimulation effects on oral force control in Parkinson's disease. AB - Dysarthria in Parkinson's disease (PD) consists of articulatory, phonatory and respiratory impairment. Bilateral subthalamic nucleus (STN) stimulation greatly improves motor disability, but its long-term effect on speech within a large group of patients has not been precisely evaluated. The aim of this study was to determine the effect of bilateral STN stimulation on oral force control in PD. We measured forces of the upper lip, lower lip and tongue in twenty-six PD patients treated with bilateral STN stimulation. Measurements of the articulatory organ force, as well as a motor evaluation using the Unified Parkinson's Disease Rating Scale (UPDRS), were made with and without STN stimulation. Maximal voluntary force (MVF), reaction time (RT), movement time (MT), imprecision of the peak force (PF) and the hold phase (HP) were all improved with STN stimulation during the articulatory force task, as well as the motor examination scores of the UPDRS. It seems that the beneficial STN stimulation-induced effect on articulatory forces persisted whatever the duration of post-surgical follow-up. However, dysarthria evaluated by the UPDRS was worse in two subgroups of patients with a one to two year and three to five year post-surgical follow-up, in comparison with a subgroup of patients with a three month follow-up. STN stimulation has a beneficial long-term effect on the articulatory organs involved in speech production, and this indicates that parkinsonian dysarthria is associated, at least in part, with an alteration in STN neuronal activity. Nevertheless, to confirm the persistence of the beneficial effect of STN stimulation on parkinsonian dysarthria, a longitudinal evaluation is still needed. PMID- 12574949 TI - Botulinum Toxin A reduces neurogenic flare but has almost no effect on pain and hyperalgesia in human skin. AB - Botulinum toxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity. There is increasing evidence that BoNT/A might also have analgesic properties, in particular in headache. In the present investigation we tested the often cited hypothesis that BoNT/A-induced analgesia can be attributed to inhibition of neuropeptide release from nociceptive nerve fibers. In 15 healthy volunteers BoNT/A (5, 10, 20 mouse units BOTOX) or saline (contralateral side) was injected intracutaneously on the volar forearm. On day zero, the day of injection, no further tests were performed. We repeatedly elicited pain, mechanical hyperalgesia and neurogenic flare by transcutaneous electrical stimulation simultaneously on the BoNT/A and saline treated side on day 1, 2, 3, 7 and 14 after injection. Before each session, sweating and local anhidrosis was assessed by iodine starch staining.BoNT/A suppressed sweating as early as from the second day after injection (p < 0.001). The size of electrically induced flare was smaller on the BoNT/A treated arm (BoNT/A side: 21.46 cm(2) +/- 3.58, saline side 24.80 +/- 3.46, p < 0.005) and BoNT/A reduced electrically-induced pain by about 10 % (p < 0.001). However, hyperalgesia to pin-prick and allodynia after electrical stimulation were unchanged. In conclusion our results indicate that peripheral neuropeptide release is attenuated by BoNT/A. In contrast, the analgesic effect of BoNT/A was very limited. Therefore we assume that other than neuropeptide mechanisms must be important for BoNT/A induced pain relief in clinical pain syndromes. PMID- 12574950 TI - Quantification of the heterogeneity of cerebral blood flow in vascular dementia. AB - BACKGROUND: In vascular dementia (VaD), assessment of cerebral blood flow by single photon emission computed tomography (CBF SPECT) has been used to detect a patchy decrease of blood flow or a frontal reduction. In addition to reduced blood flow, the heterogeneous distribution of cerebral blood flow is often observed in VaD. However, no objective method to evaluate the heterogeneity has been established. In this study, we applied three-dimensional fractal analysis (3D-FA) to CBF SPECT images as a method for assessing the heterogeneity of the cerebral blood flow distribution in VaD. SUBJECTS AND METHODS: The subjects included 18 patients with a diagnosis of VaD (aged 69.7 +/- 8.3) based on neuropsychological testing and imaging findings and 18 age-matched controls (aged 66.9 +/- 10.3). CBF SPECT images were obtained with (99m)Tc-hexamethyl propyleneamine oxime. On the reconstructed images, we obtained a linear regression equation between the cut-off values (from 35 to 50 %) and the number of voxels with a radioactivity exceeding the cut-off value transformed into natural logarithms, and then calculated the fractal dimension from the slope of the regression line thus obtained. The Mini-Mental State Examination (MMSE) was used to evaluate cognitive function. RESULTS: The fractal dimensions were 1.084 +/- 0.153 and 0.853 +/- 0.062 (mean +/- SD) in the VaD and control groups, respectively. The fractal dimension was significantly greater in the VaD group than in the control group (p < 0.0001). A significant negative correlation was observed between the fractal dimension and the MMSE score in the VaD group (r = 0.871, p < 0.0001). CONCLUSIONS: Because the CBF SPECT images of VaD patients showed a higher fractal dimension, these images were quantitatively more heterogeneous than those of age-matched controls. In the VaD group, cognitive function was shown to decline as the fractal dimension increased and images became more heterogeneous. PMID- 12574951 TI - Glaucomatous visual field defects in patients with migraine. AB - PURPOSE: To investigate the visual fields in patients with migraine in terms of glaucomatous visual field defects. METHOD: Patients who attended our Neurology outpatient clinic complaining of headache and were diagnosed as migraine were enrolled in the study. Each patient was questioned for the frequency, side, localisation and severeness of the pain as well as the onset and the duration of the symptoms, the existence of aura, the description of the aura - if present -, additional systemic problems, and family history of migraine and glaucoma. Detailed ophthalmological examinations and computerised visual field (VF) analysis were then performed on each patient in our Ophthalmology Clinic. The findings of 77 patients (61 female, 16 male) were evaluated and statistical comparisons were carried out between the group of patients with glaucomatous-like VF defects and the group without significant defects. RESULTS: VF tests revealed glaucomatous-like defects in 48 (62.3 %) patients. Intraocular pressure (IOP) levels were within normal limits in all cases. There were no statistically significant differences between the groups with and without VF defects in terms of sex, the localisation and severeness of pain, the existence and the description of aura, and family history of glaucoma, as well as the duration of migraine, IOP and cup/disc ratio. The glaucomatous group was significantly older, with a significantly higher age of onset for migraine. Risk of having glaucomatous-like defects was higher in patients who had migraine attacks once a month or less frequently. A tendency of pain and VF defects to develop ipsilaterally was noticed. DISCUSSION: A possible relationship between the pathophysiology of migraine, visual field defects and glaucomatous optic neuropathy is emphasized and VF screening for normal tension glaucoma is recommended in patients with migraine. PMID- 12574952 TI - Cognitive deficits in spinocerebellar ataxia type 1, 2, and 3. AB - Cognitive impairment was studied in distinct types of spinocerebellar ataxia (SCA): eleven SCA1, 14 SCA2, and 11 SCA3 individuals and 8 age- and IQ- matched controls. All were submitted to a neuropsychological test battery that comprised tests for IQ, attention, executive function, verbal and visuospatial memory. Executive dysfunction was prominent in SCA1 as compared with controls and all other SCA types. Mild deficits of verbal memory were present in SCA1, SCA2 and SCA3. The neuropathological pattern in different SCA types suggests that these cognitive deficits are not likely to be contingent upon cerebellar degeneration but to result from disruption of a cerebrocerebellar circuitry presumably at the pontine level. PMID- 12574953 TI - Quality of life in patients with orthostatic tremor. AB - We assessed health-related quality of life (QoL) and depression, using the SF-36 and the Beck Depression Inventory (BDI), in 20 orthostatic tremor (OT) patients. All dimensions of the SF-36 were markedly reduced in OT and depression was found in 11 patients. The BDI score correlated significantly with several SF-36 subscores. We conclude that OT strongly impacts on QoL. The results highlight the importance of recognizing and treating depression in patients with OT. PMID- 12574954 TI - Peripheral neuropathy in patients with the 3243A>G mutation in mitochondrial DNA. AB - Peripheral neuropathy is one of the clinical manifestations of the MELAS (mitochondrial encephalopathy with lactic acidosis and stroke-like episodes) syndrome, but its frequency and phenotypic variability have not been properly characterised. We therefore studied the clinical and electrophysiological features of peripheral neuropathy in 32 patients with the 3243A > G mutation in mitochondrial DNA by using clinical examination, assessment of Neuropathy Symptom Score, Neuropathy Disability Score, and electrophysiological examinations. Seven patients (22 %; 95 % confidence interval, 9-40 %) fulfilled the electrodiagnostic criteria for polyneuropathy. Mixed axon loss and demyelinating sensorimotor neuropathy was the most common type of polyneuropathy, while one patient presented with uniform demyelinating sensorimotor polyneuropathy. Sensory more than motor neuropathy was diagnosed in four patients. Clinically and electrophysiologically confirmed carpal tunnel syndrome (CTS) occurred in three patients (9.4 %), suggesting a higher prevalence than in the general population. Patients with neuropathy were in general more severely affected than those without neuropathy, although no correlation was found between the presence of neuropathy and the degree of mutant heteroplasmy in muscle. Higher age and male gender were associated with an increased risk of neuropathy. Our results show that peripheral neuropathy is not uncommon in patients with the 3243A > G mutation, and they also may have an increased risk of CTS. PMID- 12574955 TI - Accuracy of diffusion-weighted MR imaging in the diagnosis of sporadic Creutzfeldt-Jakob disease. AB - The definitive diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) is based on brain autopsy. The 14-3-3 analysis in the CSF is considered a highly sensitive and specific procedure. Sensitivity, specificity and accuracy of EEG, the 14-3-3 assay and MR imaging in 12 patients referred for suspected sCJD were calculated. We suggest that diffusion-weighted MR imaging (DWI) should be included in the array of diagnostic tests because of the 100 % sensitivity and specificity. PMID- 12574956 TI - Hemiballism with insular infarction as first manifestation of Takayasu's arteritis in association with chronic hepatitis B. AB - BACKGROUND: Takayasu's arteritis is a chronic inflammatory idiopathic disease involving large arteries like the aorta and its primary branches. Cell-mediated autoimmunity leading to vascular injury has been suspected in its pathogenesis although the antigen inducing the process remains unknown. CASE REPORT: A 50-year old male patient suffered from acute hemiballism. Neuroimaging showed an infarction of right temporal insular cortex. Neurosonology and MR-Angiography revealed bilateral long-distant subtotal stenosis of the common carotid artery and left-sided occlusion of the subclavian artery. Positive hepatitis B serology with active viral replication was found. In the absence of other vasculitis or inflammation markers, Takayasu's arteritis was diagnosed and steroid therapy was started. CONCLUSIONS: Unilateral insular lesions may lead to transient hemiballistic movements which could be the result of decreased inhibitory output of the insula to basal ganglia. The hepatitis B virus possibly contains a surface antigen inducing a specific cellular immune response leading to Takayasu's arteritis. PMID- 12574957 TI - Allogeneic stem cell transplantation after reduced-intensity conditioning in lymphoid malignancies. AB - Allogeneic stem cell transplantation (allo-SCT) is an effective therapeutic option for a wide range of hematological malignancies. The toxicity of the conditioning regimen and graft-versus-host disease (GVHD) occurring after the infusion of the graft remain the most important factors leading to high morbidity and mortality. Reduced-intensity conditioning regimens have recently been developed in an effort to reduce the toxicity associated with conventional allo SCT while preserving the curative potential of the graft-versus-tumor (GVT) effect. Most patients with lymphoproliferative disorders are not ideal candidates for allo-SCT due to higher age at diagnosis, which together with the advanced stage of disease at the time of transplantation can lead to a high transplant related mortality (TRM). Preliminary experience indicates that reduced-intensity allo-SCT is feasible in such patients. The immediate TRM is low in comparison with conventional procedures and overall results seem promising, thus indicating the existence of a GVT effect. Nevertheless, all series are still low in numbers and follow-up is too short to draw definitive conclusions. Acute and chronic GVHD remain a significant problem with incidences comparable to the conventional setting in some series. Thus, therapeutic strategies must be sought to decrease GVHD without abrogating the GVT effect. PMID- 12574958 TI - Quality of life and psychological well-being in Spanish long-term survivors of Hodgkin's disease: results of a controlled pilot study. AB - Nowadays, the chemoradiotherapeutic protocols for Hodgkin's disease (HD) achieve high curability rates. Hemato-oncologists focus on both avoiding medical and psychological sequelae of the treatment and returning patients to a normal life. The quality of life and psychological well-being of Spanish patients who are long term survivors of HD were studied and compared to the results obtained from healthy controls. Questionnaires on quality of life [European Organization for Research and Treatment of Cancer (EORTC) QLQ30] and psychological status [hospital anxiety and depression (HAD) scale] were mailed to HD patients without active disease and free of second malignancies and were also given to healthy controls. Of 67 selected patients (68.6%), 46 were included in this study. The median follow-up for these 46 patients was of 7.6 years (0.8-22.1) after being diagnosed. Although there were no differences between patients and controls with regard to their global state of health and quality of life (72.9+/-22.7 vs 79.3+/ 18.7; p=0.22), patients presented a lower physical function (88.2+/-18.1 vs 96.5+/-9.7; p=0.05) and a worse social operation scale (81.5+/-25.4 vs 96.3+/ 13.1; p= 0.0015) together with higher symptoms of dyspnea (8.6+/-14.7 vs 0+/-0; p=0.03) and higher economic difficulties (23.1+/-38.3 vs 0.7+/-4.9; p=0.017) when compared with healthy controls. However, we did not find differences in the scores and the proportion of cases of anxiety and depression between the two groups. The quality of life questionnaire disclosed differences between patients and controls in some functional and symptomatic scales. These differences can be read as a consequence of either the disease itself or the treatment received. However, the results of this controlled pilot study should be confirmed in a larger series of Spanish HD survivors. In the future, these results could be a reference when new therapeutic protocols are designed to reduce the impact on the quality of life of the patients. Socioeconomic support to the patients should also be provided in order to improve their medical care. PMID- 12574959 TI - Relationship between circulating serum soluble interleukin-6 receptor and the angiogenic cytokines basic fibroblast growth factor and vascular endothelial growth factor in multiple myeloma. AB - Angiogenesis plays an important role in multiple myeloma (MM) progression. Various mitogens such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF-2) have been implicated in the angiogenic process of various malignancies. Interleukin-6 (IL-6) is a growth factor of myeloma cells and its signaling is mediated via a cell surface receptor complex (IL-6r). IL-6 and tumor necrosis factor-alpha (TNF-alpha) are involved in the secretion of VEGF by IL-6r expressing myeloma cells. In this study, serum FGF-2, VEGF, IL-6r, and TNF-alpha were measured in 46 untreated MM patients and were studied in relation to disease stage (by Salmon-Durie criteria) and severity [assessed by serum beta(2)-microglobulin (beta(2)M), C-reactive protein (CRP), alpha(1)-antitrypsin (alpha(1)AT), and lactic dehydrogenase (LDH) levels]. The results showed that FGF 2, VEGF, IL-6r, and TNF-alpha were significantly elevated in MM patients in comparison to controls ( p<0.008) and were significantly higher in stage III disease in comparison to stages I and II ( p<0.03). The mean concentrations of IL 6r were 877+/-374, 1220+/-308, 1431+/-878, and 453+/-180 pg/ml for stages I, II, and III and controls, respectively. Levels of beta(2)M, alpha(1)AT, CRP, and LDH were all significantly higher in MM patients than controls and increased with advancing stage of disease. There were positive correlations of both VEGF and FGF 2 with IL-6r, TNF-alpha, beta(2)M, alpha(1)AT, CRP, and LDH. We conclude that IL 6r and TNF-alpha increase in parallel to VEGF and FGF-2 with increasing stage of MM disease. These molecules correlate with biochemical markers of disease activity and may play a role in the progression of multiple myeloma. PMID- 12574960 TI - Low infectious morbidity in patients with heavily pretreated hematological malignancies receiving autologous peripheral blood stem cell transplantation without antimicrobial prophylaxis. AB - The use of prophylactic antimicrobials during autologous peripheral blood stem cell transplantation (APBSCT) remains controversial. A prospective study was therefore conducted to examine whether the use of prophylactic antimicrobials is necessary. In this study, all the antimicrobials were given therapeutically rather than prophylactically. Twenty-three consecutive patients with heavily pretreated hematological malignancies were enrolled. All of the 23 patients had at least one episode of fever during APBSCT and most were fever of unknown origin (78.3%). Clinically or microbiologically documented infections occurred in only five patients (21.7%). These included bacteremias (three patients), perianal abscess (one patient), and catheter-related phlebitis (one patient). No death, invasive fungal infection, or serious adverse events occurred. The medium duration of fever, intravenous antimicrobial therapy, and hospital stay after transplantation were 5, 10, and 17 days, respectively. In conclusion, without using prophylactic antimicrobials, the infectious morbidity during APBSCT is low even in patients with heavily pretreated hematological malignancies. PMID- 12574961 TI - Platelet recovery after eradication of Helicobacter pylori in patients with idiopathic thrombocytopenic purpura. AB - The association between Helicobacter pylori ( H. pylori) infection and idiopathic thrombocytopenic purpura (ITP) has been reported by several groups. We investigated the prevalence of H. pylori infection and the effectiveness of its eradication in Japanese patients with ITP. H. pylori infection was found in 21 of 30 patients (70.0%) by (13)C urea breath test and presence of serum antibodies to H. pylori. H. pylori was eradicated in 18 of the 21 infected patients (85.7%) by administration of a proton pump inhibitor and two kinds of antibiotics. In only one patient was medication discontinued due to skin rash on the 4th day of treatment. Platelet recovery was obtained in ten patients (55.6%). In two patients with treatment failure, platelet recovery was obtained after successful re-eradication. In three patients without H. pylori infection, platelet counts did not significantly increase with the same treatment. On the other hand, eradication therapy did not affect platelet counts in patients with gastric ulcer. In conclusion, H. pylori eradication can be used for initial treatment with tolerable adverse effects in some ITP patients. PMID- 12574962 TI - Molecular characterization of deletional forms of beta-thalassemia in Taiwan. AB - beta-Thalassemia is one of the most common genetic diseases in Taiwan. The most common mutations of beta-globin are point mutations, and six mutations account for over 90% of cases. Less than 5% of the cases with beta-globin gene deletion result in beta-thalassemia minor. The mutational type of the deletion is not clear in Taiwanese. We used polymerase chain reaction (PCR)-based methods to detect the breakpoint junctions of different deletional types of beta thalassemia. In total, six cases of clinically suspected deletional type of beta thalassemia were studied. The results showed that there were three types of deletions in these cases: two cases each for hereditary persistent fetal hemoglobinemia (HPFH) of the Southeast Asian (SEA) type, HPFH of the Yunnanese type, and gamma(G)+(gamma(A)deltabeta)(0)deletions, respectively. The clinical features of these deletional mutations are milder than the beta(o) types of the point mutation. The patients with compound heterozygous mutations of the point mutation and the deletional mutation are always transfusion independent. PMID- 12574963 TI - Acute disseminated encephalomyelitis (ADEM) after allogeneic bone marrow transplantation for acute myeloid leukemia. AB - Acute disseminated encephalomyelitis (ADEM) is a rare inflammatory demyelinating disease of the central nervous system. We describe here a patient who developed ADEM after allogeneic bone marrow transplantation (BMT). A 48-year-old woman with acute myeloid leukemia (M2) underwent allogeneic BMT from her HLA-identical sister. Cyclosporin for prophylaxis of acute graft-versus-host disease (GVHD) was discontinued from day 15 because of its toxicity. She was relatively well after the resolution of cytomegalovirus reactivation and chronic GVHD. Nine months after BMT, she suddenly developed diplopia, dysarthria, and gait disturbance. Computed tomography of the brain at that time revealed no abnormal findings. Leukemia recurrence was not revealed. The neurological symptoms were very mild without further deterioration. Her clinical course was carefully watched without therapy. Two weeks after onset, fluid attenuated inversion recovery magnetic resonance imaging (MRI) revealed multifocal abnormal high-signal intensity mainly in the white matter of the cerebrum as well as in the cerebellum and brainstem. Cerebrospinal fluid examination showed no abnormal findings. No laboratory findings suggested the presence of infectious agents. The typical MRI findings and an acute monophasic clinical course of this patient led to a diagnosis of ADEM. Twelve weeks after onset, the symptoms had almost resolved. Follow-up MRI showed a substantial improvement of the previous lesions without any new lesions. The symptoms had completely resolved 5 months after onset. This is a rare case of ADEM developing after allogeneic BMT. PMID- 12574964 TI - Fatal infection caused by Francisella tularensisin a neutropenic bone marrow transplant recipient. AB - Francisella tularensis is one of the most infectious pathogenic bacteria known. Even though immunity against this organism is thought to be primarily T cell mediated, some evidence suggests that neutrophils may also play an important protective role. We report a case of tularemia in a neutropenic bone marrow transplant recipient that sheds light on the importance of neutrophils in protection against this infection and review clinical aspects of this fascinating infection emphasizing areas of interest for immunocompromised hosts. PMID- 12574965 TI - Aseptic necrosis of both femoral heads as first symptom of chronic myelogenous leukemia. AB - Chronic myelogenous leukemia (CML) is a disease of the elderly; in rare cases it occurs in childhood or adolescence. One complication at primary diagnosis is leukostasis, which usually causes respiratory, retinal, or central nervous symptoms. In this report we describe the case of a 24-year-old woman who developed aseptic necrosis of both femoral heads, which was successfully treated by bore holes in the femoral heads. This is a very rare complication of severe leukostasis, leading to the diagnosis of CML in this case. To our knowledge, this is the first case of an adult patient showing aseptic necrosis of femoral heads caused by leukostasis. PMID- 12574966 TI - Extramedullary blast crisis of chronic myeloid leukemia after allogeneic hematopoietic stem cell transplantation mimicking aggressive, translocation t(14;18)-positive B-cell lymphoma. AB - We report the case of a 42-year-old male patient who was diagnosed with a large tumor of the right thoracic aperture 30 months after unrelated hematopoietic stem cell transplantation (HSCT) for accelerated phase of Philadelphia chromosome (Ph) positive chronic myeloid leukemia (CML). Biopsy revealed an immature lymphoid neoplasia with blastic tumor cell morphology and immunoreactivity for CD34, CD79a, CD43, and CD30 as well as slight positivity for TdT and CD20. Bcr-Abl rearrangement was found in interphase tumor cell nuclei by fluorescence in situ hybridization (FISH). Furthermore, a translocation t(14;18)(q32;q21) was amplified by polymerase chain reaction (PCR). Bone marrow (BM) examination showed regular hematopoiesis including a negative FISH analysis for Bcr-Abl and complete donor chimerism. Nested PCR from peripheral blood (PB), but not conventional PCR, was positive for the b3a2 Bcr-Abl transcript. Neither radiation nor intensive chemotherapy was capable of achieving a tumor remission, and the patient died from progressive disease 6 months later. Postmortem examinations showed a shift of immunophenotype with appearance of myeloperoxidase-positive tumor cells and loss of lymphoid antigens. In addition, there were characteristic cytogenetic findings of multiple Ph chromosomes and a clonal loss of P53 tumor suppressor gene. The latter was already deleted before HSCT. We conclude that lymphoid neoplasia occurring in our patient should be interpreted as an extramedullary, very immature blast crisis of CML expressing lymphoid differentiation markers rather than a true de novo NHL. PMID- 12574967 TI - Hemophagocytic syndrome as the primary clinical symptom of Hodgkin's disease. AB - It is now well recognized that hemophagocytic syndrome (HPS) is occasionally associated with malignant lymphomas. However, its association with Hodgkin's disease has been only rarely reported. We present here a 72-year-old woman manifesting with HPS as the primary and solitary clinical symptom of Hodgkin's disease. She had been suffering from high-grade fever and anemia for more than a month. Based on the findings in bone marrow aspirates, she was diagnosed as having HPS. In spite of extensive surveys including various cultures, serological tests for collagen disease, abdominal and cardiac sonography, chest computed tomography (CT), and renal biopsy, the origin of the fever was not determined. She was treated with steroid pulse therapy and then referred. Radiological studies revealed only mild hepatosplenomegaly and small lymph node swellings around celiac and common hepatic arteries. Reevaluation of the bone marrow specimen revealed the infiltration of small numbers of CD30-, CD15-, and EBER-1 positive large-sized lymphocytes with bizarre nucleus. Under the diagnosis of Hodgkin's disease, she was treated with combination chemotherapy containing pirarubicin, cyclophosphamide, vincristine, and prednisolone. However, it was not effective and she died of rapidly progressive hepatic failure on the 5th day of the chemotherapy. Autopsy was performed, which showed proliferation of lymphoma cells in para-aortic lymph nodes. We believe that diagnostic survey to rule out the underlying lymphoma should be vigorously performed for patients with hemophagocytic syndrome of unknown origin. PMID- 12574968 TI - Auer rod-like inclusions in multiple myeloma. AB - We report a case of IgA multiple myeloma, in which the plasma cells showed multiple azurophilic, Auer rod-like intracytoplasmic inclusions in May-Grunwald Giemsa-stained marrow smears. Cytochemical stainings revealed a strong alpha-N esterase activity of these inclusions, whereas the reactions for peroxidase, Sudan black, chloroacetate esterase, and PAS were negative. Immunostaining verified IgA-kappa inside the plasma cells. The inclusions, however, were negative. Amyloid and lysozyme were also not detectable. Electron microscopy showed Auer rod-like inclusions with a smooth surface in the neighborhood of a well-developed rough endoplasmic reticulum, but with no direct relation to it. The inclusions showed a fine lamellar substructure, and the periodicity of the filamentous striations was about 10 nm, comparable with the substructure of typical Auer rods. Our findings suggest that the azurophilic inclusions in multiple myeloma are Auer rod-related structures, which likewise consist of active lysosomal enzymes. In contrast to the Auer rods in acute myeloblastic leukemia (AML), however, the inclusions in multiple myeloma consist of typical plasma cell enzymes. PMID- 12574969 TI - Cyclic thrombocytopenia and polycythemia vera. AB - A periodic fall of platelet number characterizes an acquired pathological condition named cyclic thrombocytopenia. We describe an unusual case of polycythemia vera in which the episodes of thrombocytopenia were followed regularly by thrombocytosis. The period of platelet count fluctuation was about 50 days, with the counts ranging from 34 to 820 x 10(9)/l. Bone marrow megakaryocytes were decreased in number during platelet nadir. Circulating thrombopoietin levels fluctuated out of phase with the platelet count. We suggest that at least some cases of polycythemia vera may have an unstable hematopoietic stem cell pool in nature, which could contribute to the development of unprovoked cyclic thrombocytopenia. PMID- 12574970 TI - Malignant infantile osteopetrosis initially presenting with neonatal hypocalcemia: case report. AB - Autosomal recessive "malignant" osteopetrosis is a rare congenital disorder relating to bone resorption abnormalities. It is believed to arise due to the failure of osteoclasts to resorb immature bone. This leads to abnormal bone marrow cavity formation and, clinically, to the signs and symptoms of bone marrow failure. Impaired bone remodeling associated with dysregulated activity of osteoclasts for such a condition may typically result in bony narrowing of the cranial nerve foramina, which typically results in cranial nerve (especially optic nerve) compression. Abnormal remodeling of primary woven bone to lamellar bone results in "brittle" bone that is prone to fracture. Thus, fractures, visual impairment, and bone marrow failure are the classical features of this disease. We describe the case of a 23-day-old boy in whom neonatal hypocalcemia was present initially after birth. Malignant infantile osteopetrosis (MIO) was diagnosed for the patient at 4 months of age based on evidence of anemia, thrombocytopenia, leukoerythroblastosis, sclerotic bone, hepatosplenomegaly, and visual deficit from a bony encroachment by the cranial nerve foramina. Although only occasionally reported previously, MIO remains essentially unrecognized by clinicians as a cause of neonatal hypocalcemia, which often results in diagnostic confusion and delay. This is important in the context of curative hemopoietic stem cell transplantation where preservation of sight may depend upon early intervention. PMID- 12574971 TI - Radioimmunotherapy: is avidin-biotin pretargeting the preferred choice among pretargeting methods? PMID- 12574972 TI - 99mTcO(BAT-NI), a novel nitroimidazole tracer: in vivo uptake studies in ischaemic myocardium. AB - Myocardial perfusion single-photon emission tomography (SPET) performed with cationic technetium-99m complexes indicates ischaemic areas as cold lesions. By contrast, nitroimidazole derivatives labelled with fluorine-18 or (99m)Tc have recently shown promising results for hot spot imaging of ischaemic myocardium. This study evaluates (99m)TcO(BAT-NI), a new (99m)Tc complex comprising the nitroimidazole ligand, 2,10-dimercapto-2,10-dimethyl-4,8-diaza-6-[4-(2 nitroimidazolyl)butyl]undecane, in a low-flow in vivo model of myocardial ischaemia in thoracotomised rats. To elucidate the influence of the 2 nitroimidazole group on ischaemia-induced uptake, comparisons with ligand derivatives were performed where (a) the 2-nitro group was deleted [(99m)TcO(BAT I)], (b) the 2-nitroimidazole functionality was replaced by a Br atom [(99m)TcO(BAT-Br)] and (c) the (99m)TcO(BAT) moiety was replaced by an iodine-125 iodophenoxybutyl ligand ((125)IP-NI). The radiolabelled compounds were i.v. injected 15 min after reducing resting myocardial blood flow by 50-60% and the uptake of radioactivity was assessed 90 min post injection. Autoradiography of left ventricular short-axis slices showed median uptake ratios of ischaemic/non ischaemic myocardium (I/N) of 3.4, 4.5 and 3.4 for (99m)TcO(BAT-NI), (99m)TcO(BAT I) and (99m)TcO(BAT-Br), respectively. In contrast, (125)IP-NI was not preferentially taken up by ischaemic myocardium. Accumulation of (99m)TcO(BAT-NI) in ischaemic heart regions was comparable to that in the liver. Biodistribution studies showed a median uptake of 0.65% ID/g of (99m)TcO(BAT-NI) in ischaemic tissue and an I/N of 3.3. On planar images of the thorax and upper abdomen the ischaemic hearts were visualised faintly; the median heart to lung count ratio for (99m)TcO(BAT-NI) was 1.7, and the median heart to liver count ratio was 1.0. We conclude that uptake of (99m)TcO(BAT-NI) in ischaemic myocardium does not depend on the nitroimidazole moiety but is intrinsic to the BAT complex. Clinical use of the (99m)TcO(BAT)-labelled tracers seems unlikely owing to their low uptake and their low ischaemic tissue contrast on planar images in vivo. PMID- 12574973 TI - Noninvasive imaging of isolated persistent fifth aortic arch. AB - Persistent fifth aortic arch was suspected by echocardiography and confirmed by magnetic resonance imaging (MRI) in an infant with a heart murmur. Selected images including three dimensional reconstruction from MRI demonstrating this very rare congenital anomaly are presented. PMID- 12574974 TI - Complete atrioventricular block associated with atrial flutter in fetus with positive maternal anti-SSA antibody. PMID- 12574975 TI - Clinical and electrophysiological characteristics in children with atrioventricular nodal reentrant tachycardia. AB - Atrioventricular (AV) nodal reentrant tachycardia is one of the most common supraventricular tachycardias in childhood. However, information about AV nodal reentrant tachycardia in childhood is limited, especially about the variant and multiple forms. The purpose of this retrospective study was to investigate the clinical and electrophysiological characteristics in pediatric patients with AV nodal reentrant tachycardia. Forty-eight pediatric patients with AV nodal reentrant tachycardia were included (ages 11-18 years; 25 males and 23 females). The age of onset and duration of symptoms were significantly younger and shorter in pediatric patients, respectively. A higher incidence of antegrade dual AV nodal pathways was found in adult patients than pediatric patients (72.9 vs 52.1% p = 0.003). Both antegrade and retrograde slow pathway functions were better in pediatric than adult patients. There was no significant difference between children and adults in the occurrence of variant and multiple forms of AV nodal reentrant tachycardia. This study demonstrated that pediatric patients have different electrophysiologic characteristics from those of adult patients. PMID- 12574976 TI - Interrupted aortic arch type B in trisomy 21: repair with carotid artery interposition. AB - A 21-year-old man with Trisomy 21 was diagnosed with interrupted aortic arch type B and perimembranous ventricular septal defect in the newborn period. He underwent carotid artery interposition and pulmonary artery banding with subsequent debanding and closure of the ventricular septal defect. Cardiac catheterization 20 years later demonstrated good "carotid arch" growth with no residual arch obstruction. Carotid artery interposition provided good curative repair for arch obstruction at late follow-up in this patient. PMID- 12574977 TI - Neoaortic root dilation associated with left coronary artery stenosis following arterial switch procedure. AB - We describe a patient who was diagnosed with d-transposition of the great arteries, with intact ventricular septum, who underwent arterial switch procedure on day 5 of life. Over the subsequent years, he developed progressive neoaortic root dilation with a Z score of up to 7.2. At 5 years of age, he presented with myocardial infarction. Cardiac catheterization demonstrated a markedly dilated aortic root with kinking and stenosis of the left main coronary artery into the left anterior descending coronary artery. He underwent emergency left internal mammary artery bypass grafting to the left anterior descending coronary artery. Although he required left ventricular assist device (LVAD) support in the early post-operative period, he recovered with a left ventricular ejection fraction of 52% on the most recent follow-up. PMID- 12574979 TI - Moderate-term effect of epoprostenol on severe portopulmonary hypertension. AB - We examined the effect of continuous intravenous infusion of epoprostenol (35 ng/kg/min) on severe portopulmonary hypertension caused by biliary atresia. Pulmonary hemodynamics improved and brain natriuretic peptide and human atrial natriureic peptide decreased to normal values during epoprostenol therapy. However, the improvement in pulmonary hemodynamics was not sufficient to permit liver transplantation. Our patient was obliged to stop epoprostenol therapy because of financial problems and epoprostenol was tapered off safely over 6 weeks. PMID- 12574978 TI - Concurrent transcatheter closure of an apical muscular ventricular septal defect and a patent ductus arteriosus in a child with severe hyperkinetic pulmonary hypertension. AB - Definitive treatment of congenital apical muscular ventricular septal defect (VSD) with large left-to-right shunt, severe pulmonary hypertension, and major associated lesions such as patent ductus arteriosus (PDA) has so far been possible only by surgery that has significant attendant morbidity and mortality [2]. Transcatheter device closure of both shunt lesions, if feasible, is a potentially simpler and safer alternative to surgery. PMID- 12574980 TI - A patient with kawasaki disease showing severe tricuspid regurgitation and left ventricular dysfunction in the acute phase. AB - We encountered a male aged 6 years 4 months with Kawasaki disease (KD) with severe tricuspid regurgitation and left ventricular dysfunction in the acute phase. His clinical course and findings of endomyocardial biopsy suggested that myocarditis was the cause of left ventricular dysfunction and responsible for the development of tricuspid regurgitation. With improvement of left ventricular function, the tricuspid regurgitation rapidly improved, then disappeared. There are no previous reports in the literature of KD complicated by severe tricuspid regurgitation. PMID- 12574981 TI - Familial truncus arteriosus: a possible autosomal-recessive trait. AB - Truncus arteriosus is a relatively uncommon congenital malformation. It accounts for approximately 1% of congenital heart diseases. The defect occurs sporadically but 22q11 deletion is frequently noted in such patients with conotruncal defects. We studied six cases of TA in four closely related families. Analysis of karyotypes in these cases was normal. Family 1 has one affected male infant who was born in 1998. Family 2 has two affected children (one male and one female) who were born in 1989 and 1995, respectively. They have four other normal children. Family 3 has two affected children (one male and one female) who were born in 1981 and 1984, respectively. They have three other normal children. Family 4 has one affected male born in 1998 and another healthy child. All parents of all affected children are double cousins. The data in this study are compatible with an autosomal-recessive inheritance, but multifactorial inheritance may also play a role. PMID- 12574982 TI - Proximal left coronary artery occlusion in a 15-year-old boy with noonan syndrome and hypertrophic cardiomyopathy. AB - Whereas intra- and extramural coronary artery narrowing have been found in patients with hypertrophic cardiomyopathy, occlusion of extramural coronary arteries has not been reported. Also, only three cases with Noonan syndrome and congenital coronary artery anomalies have been described. We report a patient with Noonan syndrome, hypertrophic cardiomyopathy, and complete occlusion of the proximal left coronary artery. PMID- 12574983 TI - Congenital long QT syndrome and 2:1 atrioventricular block with a mutation of the SCN5A gene. AB - Infants with congenital long QT syndrome (LQTS) and 2:1 atrioventricular block (AVB) have been recognized as a clinical subset of children with LQTS. However, the genotype of this disorder is not well-known. We report an infant with LQTS and 2:1 AVB with a mutation of the SCN5A gene (LQT3). In some patients with LQTS and 2:1 AVB, the disorder may be due to mutation of the SCN5A gene (LQT3). PMID- 12574985 TI - Intraoperative transesophageal echocardiography in a 1.4-kg infant with complex congenital heart disease. AB - The lower weight limit for infants undergoing intraoperative transesophageal echocardiography (TEE) with current commercially available probes has not been determined. A review of the literature reveals that infants as small as 1.6 kg have been studied successfully. This report describes the first intraoperative TEE reported in a 1.4-kg infant during truncus arteriosus/interrupted aortic arch repair. Successful pre- and postoperative images of the cardiac abnormality were obtained. Probe insertion was performed in this small patient after predilating the esophagus with a 14-F suction catheter. PMID- 12574984 TI - Pulmonary artery calcification in recipient twins of twin to twin transfusion syndrome: a report of three cases. AB - Twin to twin transfusion syndrome may rarely complicate monochorionic twin gestation, resulting in discordant size of the twins. Cardiac dysfunction and various types of cardiac defects have been described in the recipient twin of this syndrome. Pulmonary artery calcification has been described earlier in one case only. We report three recipients of twin to twin transfusion syndrome with pulmonary artery calcification resulting in obstruction of pulmonary trunk in two of these. One case with progressive severe narrowing was successfully operated at four months of age with patch repair of pulmonary artery. He is doing well three years later. The obstruction is moderate in the other case. Calcification in the wall of pulmonary artery may be secondary to vascular injury sustained as a result of volume overloading of the circulation of the recipient twin. PMID- 12574986 TI - Jarcho-levin syndrome associated with a complex congenital heart anomaly. AB - Jarcho-Levin syndrome is characterized by "crab-like" rib cage deformity and multiple vertebral anomalies that cause respiratory failure. Reports of complex congenital heart defects with this syndrome are rare. We describe a female infant with this syndrome and a complex congenital heart defect and review the literature. Congenital heart defects are occasionally complicated by Jarcho-Levin syndrome. Heart defects involving heterotaxic morphology should be included as one of the important clinical features of this syndrome. PMID- 12574990 TI - Caffeine--a drug with a surprise. PMID- 12574991 TI - How unique is the genetic code? PMID- 12574987 TI - Large differences between LINE-1 amplification rates in the human and chimpanzee lineages. AB - The genomic evolution and causes of phenotypic variation among humans and great apes remain largely unknown, although the phylogenetic relationships among them have been extensively explored. Previous studies that focus on differences at the amino acid and nucleotide sequence levels have revealed a high degree of similarity between humans and chimpanzees, suggesting that other types of genomic change may have contributed to the relatively large phenotypic differences between them. For example, the activity of long interspersed element 1 (LINE-1) retrotransposons may impose significant changes on genomic structure and function and, consequently, on phenotype. Here we investigate the relative rates of LINE-1 amplification in the lineages leading to humans, bonobos (Pan paniscus), and chimpanzees (P. troglodytes). Our data indicate that LINE-1 insertions have accumulated at significantly greater rates in bonobos and chimpanzees than in humans, provide insights into the timing of major LINE-1 amplification events during great ape evolution, and identify a Pan-specific LINE-1 subfamily. PMID- 12574992 TI - Application of the duality principle to chiral icosahedral metal complexes. PMID- 12574993 TI - Biomineralization of unicellular organisms: an unusual membrane biochemistry for the production of inorganic nano- and microstructures. AB - With evolution, Nature has ingeniously succeeded in giving rise to an impressive variety of inorganic structures. Every organism that synthesizes biogenic minerals does so in a form that is unique to that species. This biomineralization is apparently biologically controlled. It is thus expected that both the synthesis and the form of every specific biogenic mineral is genetically determined and controlled. An investigation of the mechanism of biomineralization has only become possible with the development of modern methods in molecular biology. Unicellular organisms such as magnetic bacteria, calcareous algae, and diatoms, all of which are amongst the simplest forms of life, are particularly suited to be investigated by these methods. Crystals and composites of proteins and amorphous inorganic polymers are formed as complex structures within these organisms; these structures are not known in conventional inorganic chemistry. PMID- 12574994 TI - A layered nickel succinate with unprecedented hexanickel units: structure elucidation from powder-diffraction data, and magnetic and sorption properties. PMID- 12574996 TI - Synthesis of porphyrin-containing [3]rotaxanes by olefin metathesis. PMID- 12574995 TI - Towards the development of colorimetric probes to discriminate between isomeric dicarboxylates. PMID- 12574997 TI - Alkali-metal-controlled self-assembly of crown-shaped ring complexes of lanthanide/[alpha-AsW9O33]9-: [K subset [Eu(H2O)2(alpha-AsW9O33)]6]35- and [Cs subset [Eu(H2O)2(alpha-AsW9O33)]4]23-. PMID- 12574998 TI - Facile solid-state synthesis of highly conducting poly(ethylenedioxythiophene). PMID- 12574999 TI - Highly enantioselective organocatalytic conjugate addition of malonates to acyclic alpha,beta-unsaturated enones. PMID- 12575000 TI - Supramolecular chirality: a reporter of structural memory. PMID- 12575001 TI - Triboracyclobutanides: four-membered two-electron aromatic compounds with fluctuating skeletal bonds. PMID- 12575002 TI - A diboracyclopropane with a planar-tetracoordinate carbon atom and a triborabicyclobutane. PMID- 12575003 TI - Long-range electronic coupling in various oxidation states of a C4-linked tris(beta-diketonato)ruthenium dimer. PMID- 12575004 TI - all-D-Polypeptides: novel targets for semisynthesis. PMID- 12575005 TI - Photoinduced intrarotaxane electron transfer between zinc porphyrin and [60]fullerene in benzonitrile. PMID- 12575006 TI - Isomeric constellations of encapsulation complexes store information on the nanometer scale. PMID- 12575007 TI - Modular cavity-tunable self-assembly of molecular bowls and crowns as structural analogues of calix[3]arenes. PMID- 12575008 TI - Structure and reactivity of a mononuclear gold-complex catalyst supported on magnesium oxide. PMID- 12575009 TI - Total synthesis of (-)-spirotryprostatin B. PMID- 12575011 TI - Influence of maternal carbohydrate intake on fetal movements at 14 to 16 weeks of gestation. AB - OBJECTIVE: Ultrasonographic assessment of fetal movements at 14 to 16 weeks of gestation before and after maternal oral intake of carbohydrate. DESIGN: Thirty consecutive healthy pregnant women between 14 and 16 weeks of gestation were scanned transvaginally. Real-time ultrasound recordings of 20 min duration were performed in the fasting state and after an oral intake of 110 g of glucose. The number of fetal movements, body and limb movements and breathing movements were evaluated. Fetal movements were quantified using a digital chronometer, and the percentage of time spent moving and the rate of movements per minute were then calculated. Data was statistically analyzed using the simple and Wilcoxon paired t-test. RESULTS: Absolute number of fetal movements, body movements, rotation movements, time spent moving and rate of movements per minute showed a statistically significant increase after maternal intake of carbohydrate (p < 0.0001). CONCLUSIONS: Maternal oral intake of 110 g of carbohydrate significantly increases fetal movements at 14 to 16 weeks of gestation, thus allowing better ultrasonographic viewing of the fetus. PMID- 12575012 TI - Maternal serum marker medians in Aboriginal Canadian women. AB - OBJECTIVES: The study evaluates the differences between Aboriginal and Caucasian women in the levels of maternal serum markers used in second-trimester Down syndrome screening (alpha-fetoprotein, unconjugated estriol, and total human chorionic gonadotrophin). METHODS: A case-control study compared the levels of serum markers in 401 Aboriginal women and 1565 matched controls selected from 7717 Caucasian women. The cases and controls were screened in a single centre and matched for maternal age, parity, and sample date. Women with multiple pregnancies and pregnancies associated with Down syndrome, open neural tube defects, trisomy 18, and insulin-dependent diabetes mellitus as well as women without weight recorded were excluded from the study. RESULTS: No differences in the levels of maternal serum alpha-fetoprotein and total human chorionic gonadotrophin were observed between the two groups. Maternal serum unconjugated estriol was 12% higher in Aboriginal women. DISCUSSION: Since Aboriginal women make up only a small proportion of women screened, correcting the level of uE3 for this group will have little effect on the overall screening performance. However, if these results are confirmed by further study, individual centres may consider making this correction, so optimal screening performance can be achieved in Aboriginal women. PMID- 12575013 TI - Tetrasomy 12p--unusual presentation in CVS. AB - CVS direct preparations usually achieve limited resolution and are better at detecting numerical rather than structural abnormalities. A CVS direct preparation analyzed using G-banding revealed a 47,XY,+G karyotype in 5 of 11 cells and was reported as mosaic for trisomy 21. Subsequent analysis of the CVS culture found only normal male cells. Amniocentesis revealed both normal male cells and cells with an extra F-group chromosome. Fluorescence in situ hybridization (FISH) identified this chromosome to be an isochromosome from the short arm of chromosome 12 [i(12)(p10)]. The amniocyte karyotype was reported as 47,XY,+i(12)(p10)[12]/46,XY[8].ish i(12)(p10)(wcp12+), which is associated with Pallister-Killian syndrome. Reexamination of the CVS direct preparation by FISH with a chromosome 12 centromere probe confirmed the karyotype of this tissue to be 47,XY,+mar[5]/46,XY[6].nuc ish 12cen(D12Z3 x 3)/12cen(D12Z3 x 2). Thus, multiple studies, including amniocentesis and fluorescence in situ hybridization, may be required to fully and accurately evaluate abnormalities detected by CVS. This case also indicates that mosaicism for supernumerary isochromosomes may have a complex origin. PMID- 12575014 TI - Intrauterine brain teratoma: a case report of imaging (US, MRI) with neuropathologic correlations. AB - Fetal brain tumors are rare and teratoma is considered as the most common. Fetal MR Imaging is currently used to evaluate cases of ventricular dilatation. We report a case of cerebral immature teratoma detected by ultrasonography because of ventricular dilatation at 24 gestational weeks. MRI was the more accurate imaging method in depicting the tumor and its consequences on brain development as well as in taking a decision with regard to the management of pregnancy. PMID- 12575015 TI - Prenatal diagnosis of dilated coronary sinus with persistent left superior vena cava in a fetus with trisomy 18. AB - A case of dilated coronary sinus with persistent left superior vena cava diagnosed at 33 weeks in a fetus with trisomy 18 is reported. The features of this cardiac anomaly on prenatal ultrasonography and its association with trisomy 18 are discussed. Published in 2003 John Wiley & Sons, Ltd. PMID- 12575016 TI - Circulating trophoblast in maternal blood. AB - This review describes the status of circulating trophoblast, but is considered in the perspective that only a specific subset of trophoblast cells circulates in the maternal blood. The consequences for isolation, identification and clinical potential are described. PMID- 12575017 TI - Antenatal detection of a single umbilical artery: does it matter? AB - The presence of a single umbilical artery is recognised as a soft marker for congenital anomalies, aneuploidy, earlier delivery and low birthweight. Most of the available data are derived from case series or highly selected populations and are therefore likely to be unrepresentative. In this retrospective case comparison study, we firstly aimed to determine the incidence of a single umbilical artery in an unselected population and secondly to examine the clinical significance of this soft marker. Over a 40-month period, 107 cases were identified from a cohort of 35 066 births giving an incidence of 3.1 per 1000 total births and late pregnancy losses. The antenatal detection rate was only 30%. Compared to fetuses with normal cord vasculature, fetuses with a single umbilical artery were more likely to be delivered at an earlier gestation and to weigh less, were 1.7 times more likely to be delivered by a Caesarean section and 19% of the cases had a congenital anomaly. The perinatal mortality was 49.0 per 1000 total births, which was 6 times higher than the background hospital rate. The presence of a single umbilical artery is associated with a poorer perinatal outcome compared to that in fetuses with three vessels in the cord. Unfortunately, the antenatal detection rate is poor. Recognising the importance of this soft marker in counselling and management of pregnancies should provide the stimulus to improve detection rates. PMID- 12575018 TI - Prenatal diagnosis of occipital encephalocele, mega-cisterna magna, mesomelic shortening, and clubfeet associated with pure tetrasomy 20p. AB - We present the first case of a fetus with pure tetrasomy 20p proven by cord-blood sampling at 24 weeks of gestation. This case was diagnosed in utero with multiple congenital anomalies including occipital encephalocele, mega-cisterna magna, mesomelic shortening, and clubfeet. An analysis of GTG-banded chromosomes of 20 metaphase cells was performed. Female karyotype [47,XX, +i(20)(p10)] was revealed in all cells. Pure tetrasomy 20p was confirmed using fluorescent in situ hybridization (FISH) with a telomere probe for chromosome 20p in all seven metaphase cells. The pregnancy was terminated because of associated multiple anomalies and severe oligohydramnios. The postmortem examination confirmed the prenatal diagnosis. PMID- 12575019 TI - Prenatally detected trisomy 4 and 6 mosaicism--cytogenetic results and clinical phenotype. AB - We report on a live-born male with 46,XY/47,XY+4/47,XY,+6 mosaicism. Trisomy 4 mosaicism was detected by karyotyping chorionic villus samples (CVS) and was confirmed by the analysis of 16 metaphases obtained from cultured amniotic fluid cells. Eight metaphases were normal (46,XY), two had trisomy 4 (47,XY,+4), and two had trisomy 6 (47,XY,+6). Two postnatal chromosomal analyses of blood lymphocytes at birth and at the age of one week were normal. Chromosomal analysis of cultured skin fibroblasts from the right inguinal region at the age of 12 months revealed trisomy 4 (47,XY,+4) in 49 metaphases, trisomy 6 (47,XY,+6) in 2 metaphases, and a normal karyotype (46,XY) in 49 cells of the 100 analyzed metaphases, respectively. The main clinical findings consist of prenatal growth retardation, hypoplasia of the right side of the face, a dysplastic and posteriorly rotated right ear, a high vaulted palate, retrognathia, aplasia of the right thumb, hypoplasia of the fingernails, a deep sacral dimple, and patchy skin hypopigmentation of the right leg. When last seen at the age of 14 months, his development was nearly normal. Five patients with trisomy 4 mosaicism have been reported previously, but none with an additional trisomy 6 mosaicism. PMID- 12575020 TI - Prenatal diagnosis of holoprosencephaly in two fetuses with der (7)t(1;7)(q32;q32)pat inherited from the father with double translocations. AB - The presence of two independent translocations in one person is rare. Herein, we report the prenatal diagnosis of two sibling fetuses with holoprosencephaly, whose father is a carrier of double translocations. The karyotype of the father is 46,XY, t(1;7) (q32;q32), t(14,15) (q32.1;q26.3). The two fetuses had variable facial dysmorphisms and identical cytogenetic abnormality-a derivative (7) t(1;7) (q32;q32) inherited from the father. The proband 1 showed a small mouth, a single median eye and a proboscis above the eye, while the proband 2 showed hypotelorism, a flat nose, cleft lip and cleft palate. Both fetuses also had alobar holoprosencephaly. Haploinsufficiency of the sonic hedgehog gene at 7q36 does account for the occurrence of holoprosencephaly in the two fetuses with a deletion of distal 7q (7q32 --> qter). PMID- 12575021 TI - First-trimester diagnosis of Bartsocas-Papas syndrome (BPS) by transvaginal ultrasound: case report and review of the literature. AB - Initially described in 1972, Bartsocas-Papas syndrome (BPS) is an autosomal recessively inherited disorder combining multiple pterygia, ankyloblepharon, cleft lip and palate, filiform bands between the jaws, syndactyly, and other anomalies. Although described as lethal, review of the literature reveals three individuals who survived into childhood with this condition. We describe a fourth surviving patient and what we believe to be the first prenatal diagnosis of BPS in the first trimester. PMID- 12575022 TI - Prenatal diagnosis of a small supernumerary, XIST-negative, mosaic ring X chromosome identified by fluorescence in situ hybridization in an abnormal male fetus. AB - Marker or ring X [r(X)] chromosomes of varying size are often found in patients with Turner syndrome. Patients with very small r(X) chromosomes that did not include the X-inactivation locus (XIST) have been described with a more severe phenotype. Small r(X) chromosomes are rare in males and there are only five previous reports of such cases. We report the identification of a small supernumerary X chromosome in an abnormal male fetus. Cytogenetic analysis from chorionic villus sampling was performed because of fetal nuchal translucency thickness and it showed mosaicism 46,XY/47,XY,+r(X)/48,XY,+r(X),+r(X). Fluorescence in situ hybridizations (FISH) showed the marker to be of X chromosome origin and not to contain the XIST locus. Additional specific probes showed that the r(X) included a euchromatic region in proximal Xq. At 20 weeks gestation, a second ultrasound examination revealed cerebral abnormalities. After genetic counselling, the pregnancy was terminated. The fetus we describe is the first male with a mosaic XIST-negative r(X) chromosome identified at prenatal diagnosis. The phenotype we observed was probably the result of functional disomy of the genes in the r(X) chromosome, secondary to loss of the XIST locus. PMID- 12575023 TI - Rapid prenatal diagnosis of Down syndrome using quantitative fluorescence in situ hybridization on interphase nuclei. AB - OBJECTIVES: Presently, conventional cytogenetic analysis of metaphase chromosomes remains the reference approach in prenatal diagnosis. However, this method is labor-intensive and time-consuming. The first step toward the rapid identification of aneuploidies is achieved by interphase fluorescence in situ hybridization (FISH) with centromeric or locus-specific probes. Spot counting using this type of probes is a reliable approach, but is very time-consuming with some technical and biological limitations. In this study, we present a new FISH method using image cytometry for the detection of trisomy 21 within interphase nuclei. METHODS: The method is based on a comparative quantitation of the fluorescence signals emitted by whole chromosome 21 and 22 painting probes cohybridized on interphase nuclei. The chromosomal imbalance was determined with an automated image cytometer by detecting an abnormal ratio of both fluorescence emissions when compared with the ratio obtained in normal cells. RESULTS: Ten blood samples and twenty amniotic fluids were analyzed. Results from FISH and standard cytogenetics were compared and 100% correlation was achieved. CONCLUSIONS: This method, which enables an easy detection of chromosomal imbalances without a need for metaphase preparations, can be applied to the diagnosis of trisomy 21 and extended to other disorders with chromosomal imbalances. Compared to other interphase FISH techniques, it avoids spot-scoring difficulties. PMID- 12575024 TI - Fetal supraventricular tachycardia: a role for amiodarone as second-line therapy? AB - OBJECTIVE: The aim of this study was to evaluate the role of amiodarone for the prenatal treatment of hydropic fetuses with supraventricular tachycardia. METHODS: A group of 26 hydropic fetuses with supraventricular tachycardia was studied retrospectively. RESULTS: Twenty-five fetuses received transplacental treatment. The overall prenatal conversion rate was 60%. Nine fetuses were converted to sinus rhythm using either flecainide (n = 7) or amiodarone (n = 2) as first line therapy, whilst digoxin alone or in association with sotalol failed to restore sinus rhythm in all cases. After first-line therapy, supraventricular tachycardia persisted in 10 fetuses. Nine fetuses received amiodarone alone or in association with digoxin as second-line therapy, five of whom were converted to sinus rhythm. Among the 11 live neonates treated by amiodarone in utero, 2 (17%) presented an elevated thyroid stimulating hormone at day 3-4. These two infants received thyroid hormone substitution therapy and had a normal outcome. CONCLUSION: When first-line therapy fails to restore sinus rhythm in hydropic fetuses with supraventricular tachycardia, amiodarone therapy should be considered as it allows a substantial number of fetuses to be converted prenatally. PMID- 12575025 TI - First-trimester maternal serum PAPP-A, SP1 and M-CSF levels in normal and trisomic twin pregnancies. AB - OBJECTIVE: To study PAPP-A and SP1 for biochemical trisomy screening in twin pregnancies and to investigate the role of maternal and placental compartments in marker production by comparing the levels of the decidual cytokine M-CSF with the PAPP-A and SP1 from the placenta. METHODS: Thirteen twin pregnancies with at least one chromosomally abnormal fetus were compared with 68 normal twin pregnancies. Sera were obtained between 11 + 3 and 13 + 6 weeks of gestation, and PAPP-A, SP1 and M-CSF levels were determined by immunoassay. These concentrations were also compared with gestation-matched groups of 18 singleton normal pregnancies and 18 singleton Down syndrome pregnancies. RESULTS: PAPP-A and SP1, but not M-CSF, levels were higher in normal twin pregnancy than in normal singleton pregnancy. SP1 levels, but not PAPP-A, correlated to M-CSF. PAPP-A, but not SP1, levels were reduced in abnormal twin pregnancies, with an increasing effect according to the number of affected fetuses, and were more pronounced in pregnancies with trisomy 18 or 13 than in trisomy 21 fetuses. M-CSF was inconsistent, with a trend towards increased levels in trisomy 21. CONCLUSION: PAPP-A remains the best biochemical screening marker for fetal trisomies 21, 18 or 13, in singleton as well as in twin pregnancy. In contrast to SP1, its site of production is not likely to be restricted to the placenta. The role of the (maternally produced) M-CSF remains to be further investigated. PMID- 12575026 TI - Case report: a prenatal case of Jarcho-Levin syndrome diagnosed during the first trimester of pregnancy. AB - The Jarcho-Levin syndrome is a specific form of spondylocostal/spondylothoracic dysostosis. There have been various classifications of this syndrome. We present the case of a severe prenatal Jarcho-Levin syndrome, diagnosed by ultrasound examination during the first trimester of pregnancy in a family with no previous medical history of an affected child. X-ray exploration, high-resolution spiral computed tomography and autopsy confirmed the diagnosis. PMID- 12575027 TI - Midtrimester triple-test levels in women with chronic hypertension and altered renal function. AB - OBJECTIVE: To study the association between midtrimester triple-test levels and renal function in pregnant women with chronic hypertension. METHODS: Forty-three patients with chronic hypertension for whom renal function was evaluated by the creatinine clearance test between 14- and 20-weeks gestation were studied. Midtrimester triple-test values obtained as part of Down screening program were correlated with renal function. RESULTS: hCG correlated inversely with creatinine clearance (r = -0.345; p = 0.002) and directly with alpha-fetoprotein (r = 0.499; p = 0.003). CONCLUSIONS: In pregnant women with chronic hypertension, impaired renal function may alter midtrimester maternal serum hCG levels. PMID- 12575028 TI - Fetal cells in cervical mucus in the first trimester of pregnancy. AB - OBJECTIVES: The aim of this study was to first evaluate the presence of fetal cells in cervical mucus samples collected in the first trimester of pregnancy and then to compare different laboratory methods for the detection of these cells. METHODS: Mucus samples were collected by using a cytobrush before termination of pregnancy (TOP) from 143 pregnant women between 7 and 12 weeks of gestation. None of the women had undergone an invasive diagnostic procedure prior to cervical mucus sampling. Samples of placental tissue were collected from each patient at TOP. Slides from each sample were first observed under an inverted microscope to detect possible sperm contamination. In the first part of our experiments, 40 mucus samples were treated with a mucolytic solution containing N-acetylcysteine (AC) and were analysed by a polymerase chain reaction (PCR) assay. The second series, consisting of 71 mucus samples, was treated with a mucolytic solution containing dithiothreitol (DTT): all 71 samples were analysed by a PCR-based assay, and an aliquot for fluorescent in situ hybridisation (FISH) analysis was also obtained from 48 out of 71 samples. In the third part of our experiments, performed on 32 mucus samples, mucus trapped on the cytobrush was directly spread on two slides for FISH analysis without any mucolytic treatment. All placental tissue samples obtained at termination were analysed by FISH for fetal sexing. RESULTS: Overall, the use of PCR-based or FISH analyses on 143 mucus samples resulted in correct sex prediction in 92/143 (64.3%) samples [20/66 (30.3%) cases from known male pregnancies and 72/77 (93.5%) cases from known female pregnancies]. In the AC group, Y-derived sequences were found in 7/23 samples (30.4%) from known male pregnancies and in 1/17 cases from known female pregnancies, with an overall correct sex prediction in 23/40 cases (57.5%). In the DTT group, Y-derived sequences could be amplified in 10/30 samples (33.3%) from known male pregnancies and in 4/41 cases from known female pregnancies, with an overall correct sex prediction in 47/71 cases (66.2%). In the DTT samples analysed by FISH, nuclei bearing XY signals were detected in 5/26 (19.2%) cases from known male pregnancies and in none from female pregnancies, the rate of correct sex prediction being 56.2% (27/48). On untreated mucus samples analysed by FISH, nuclei with XY signals were documented in 3/13 (23%) samples from male conceptuses and in none from known female pregnancies, with an overall correct sex prediction in 22/32 cases (68.7%). CONCLUSION: Fetal cells were not detected in a constant and reliable fashion in cervical mucus samples collected in the first trimester of pregnancy. The detection rate was poorly influenced by the use of different laboratory methods. This sampling technique cannot be regarded as a promising tool towards minimally invasive prenatal diagnosis. PMID- 12575029 TI - Prenatal diagnosis of a triplet pregnancy complicated with one acardius in a patient with didelphys uterus using three-dimensional ultrasound. PMID- 12575030 TI - Prenatal skin edema mimicking a recurrence of meningo-encephalocele. PMID- 12575031 TI - Prenatal identification of a G338E mutation in FGFR2 in a fetus without sonographic appearance of craniosynostosis. PMID- 12575032 TI - Hyperglycosylated hCG cannot be measured using a sialic acid-specific lectin immunoassay. PMID- 12575034 TI - [Affection of exogenous gibberellic acid (GA3) on endogenus hormones of Panax quinquefolium seed during its morphological after ripening period]. AB - According to the determination of endogenus hormones variation of Panax quinquefolium seed during its morphological afterripenging period, and the affection of exo-GA3 on the endogenus hormones dynamics, it suggested that exo GA3 couldn't be used for accelerate the growth of Panax quinquefolium embryo, but was helpful in relieving seed dormancy during physiological afterripening period. PMID- 12575035 TI - [The pharmacogonostical identification of Asteropyrum cavaleriei]. AB - The descriptions and microscopy identification of Asteropyrum cavaleriei were studied and compared with A. poltatum. The morphological and histological characteristics of the medicinal plants from the genus was presented. PMID- 12575036 TI - [The descriptions identification of "yimu cao"]. AB - Remained but entire bracts beneath the inflorescence and the persistent calyxes were selected as primary characteristics to distinguish "Yimu Cao", which derived from 8 species and 1 variety in the same genus Leonurus. The results were reliable. PMID- 12575037 TI - [Quality evaluation of indigowoad root and leaf by chemical pattern recognition]. AB - 27 samples of indigowoad root and 5 samples of indigowoad leaf from different species and in different habital were collected. The chemical constituents were analyzed by HPLC, and ten original chemical features as a whole obtained. Meanwhile, extrinsic bacteriostasis experiments were carried out for each of the 32 samples. Canonical correlation analysis was applied to the intercorrelation between the chemical and the pharmacologic data, and five chemical features were found to be the effective constituents. In the end, the 32 samples were divided into five classes by the fuzzy clustering technique ISODATA(interachive self organizing data analysis technique A). PMID- 12575038 TI - [Supercritical extraction of Alpinia officinarum and GC-MS analysis of its components]. AB - In order to find the effective components and new pharmaceutical effect of Alpinia officinarum, and further exploit it, we extracted its essential oils with the technique of Supercritical-CO2 fluid, then confirmed there were 27 components in Pot I And 111 components in Pot II by using the chemical component analysis and the computer information retrieval in the GC-MS system. PMID- 12575039 TI - [Studies on Euphorbia fischeriana diterpenoide lactones inhibitory effect on human tumor cells in vitro]. AB - In this paper, two diterpenoide lactones 16-hydroxypseudojolkinolide B and jolkinolide B were isolated from Euphorbia fischeriana Steud. They showed inhibitory effect on human tumor cells both the K562 cell and CNE2 cell and significant dose-effect relationship. The IC50 values of 16 hydroxypseudojolkinolide B on the two human tumor cells were near to that of ADM's. PMID- 12575040 TI - [Effect of AMI on proliferative cycle phase of bone marrow cells in mice]. AB - By using culture of bone marrow cells in vitro and flowcytometry, effect of Astragalus membranaceus injection (AMI) on proliferative cycle phase of bone marrow cells in normal and anemic mice was studied. AMI 40 micrograms/ml (concentration in culture system) can promote normal murine bone marrow cells (BMC) entering proliferative cycle phase (S + M/G2 phase), so do AMI 40 micrograms/ml and AMI 400 micrograms/ml to anemic murine BMC. The results suggested AMI maybe enhance hematopoietic function in mice. PMID- 12575041 TI - [The effects of starfish sterol on platelet aggregation]. AB - Through the internal and external experiments of platelet aggregation on rats and rabbits, we study the effects of starfish sterol (No. A1998) on the formation of experimental thrombus in vitro and the effects of platelet aggregation induced by arachidonic acid (AA), ADP, CaCl2. Starfish sterol A 1998 can inhibit the formation of experimental thrombus in vitro of rats on the rates of 10.0%, 20.1%, 25.9% (P < 0.01), p.o. starfish sterol 4.5, 13.5, 40.5 mg/kg. A 1998 can inhibit platelet aggregation induced by AA, ADP, CaCl2 in the same time. It can be concluded that starfish sterol has inhibitory effects on platelet aggregation. PMID- 12575042 TI - [Pharmacological studies of trilex on treatment of pharyngitis]. AB - Trilex consisted of three species: Ilex latifolia, Ilex asprella and Ilex rotunda. Trilex had the antipyretic effects, decreased the rat body temperature about 0.8 degree C on yeast induced hyperthermia. Trilex could obviously enhance the threshold of hot plate induced pain in mice. The maximum threshold increased 43.8%. Trilex could also inhibit acetic acid induced inflammatory ooze in mice abdominal cavity. The inhibitory rate was 34.8%. These results showed that trilex had antipyretic, analgesic and anti-inflammatory effects. It could be used to treat acute and chromic pharyngitis. PMID- 12575043 TI - [A new cultivated technique of Rehmannia glutinosa by sprout cutting]. AB - The traditional cultivated method of Rehmannia glutinosa is the asexual reproduction that small rhizomes are used as materials of reproduction. In this test, the small rhizomes germinated on seedbed, then young sprouts were picked for culture. PMID- 12575044 TI - [Investigation and identification of "shihu" medicinal materials from Shanghai]. AB - "Shihu" medicinal materials from Shanghai were investigated, colrected and identified. The results showed that the main varietis are the plants from Dendrobium, the small number from Flickingeria and Pholidota. PMID- 12575045 TI - [Morphological and histological identification of Desmodium styracifolium]. AB - The histological structures of stem and leaf for Desmodium styracifolium and the power micro-characteristics of its herba were studied. The results could supply basis for the identification of the medicinal materials. PMID- 12575046 TI - [Morphological and histological studies of Herba Boschniakiae]. AB - This paper deals with the morphological characters of the plants and drugs, and the histological characters of the stems of Herba Boschniakiae (Boschniakia rossica). This results provide authentic methods for the identification of Herba Boschniakiae. PMID- 12575047 TI - [Studies on the chemical constituents of soft coral Sarcophyton elegans]. AB - The studies on the chemical constituents of soft coral Sarcophyton elegans led to the isolation of gorgosterol, batyl alcohol and simumeramide. Their structures were identified on the comprehensive spectral analysis. These compounds were first obtained from the soft coral. PMID- 12575048 TI - [GC-MS analysis of essential oil from the leaves of Psidium guajava]. AB - The constituents of essential oils from the leaves of Psidium guajava Linn were analyzed by GC-MS qualitatively and quantitatively. Sixty compounds of the essential oils were identified at rate 90.56%. The major components were caryophyllene (18.81%), copaene (11.80%), [1aR-(1a alpha, 4a alpha, 7 alpha, 7a beta, 7b alpha)]-decahydro-1,1,7-trimethyl-4-methylene-1H-cycloprop[e] azulene(10.27%), eucalyptol(7.36%). PMID- 12575049 TI - [Experimental studies on pharmacodynamic effect of Macleaya cordata]. AB - The pharmacodynamic effect of Macleaya cordata was investigated in this study. The results showed that Macleaya cordata could improve liver function of acute hepatic injuries in rats caused by tetrachloromethan or galactoslmine. Macleaya cordata could lessen the level of serum LDH and mortality of rats, increase ratio of serum A/G, protect cellular membrane efectively and inhibit fibrosis in rats with chromic heptic injury caused by tetrachloromethane. Macleaya cordata still enhanced the function of T and B lymphocytes. PMID- 12575050 TI - [Antitumor activity of the root of Euphorbia helioscopic in vitro]. AB - Antitumor activity of the aquatic extract the root of Euphorbia helioscopia L (EWE) in Vitro were studied. Viable cells count, MTT staining and colonal formation assays of three kinds of cancer cells were used to assess the antitumor activity. Determined by viable cells count, the IC50 values of EWE against 7721, Hela, MKN-45 cells were 1.26, 1.98, 1.72 mg/ml respectively (72 h). Determined MTT staining, the IC50 values EWE against 7721, Hela, MKN-45 cells were 1.43, 1.67, 0.97 mg/ml. Determined by colonal formation, the inhibition rate of EWE (4 mg/ml) against 7721, Hela, MKN-45 cells were 59.8%, 66.4%, 70.5%. The results indicated that EWE had obvious antitumor activity. PMID- 12575051 TI - [Effects of abalong polysaccharide on the activity of the peritoneal macrophages and delayed-type hypersensitivity in mice bearing S180]. AB - The effect of Abalone polysaccharide(Ap) which extracted from Haliotis discus hannai Ino on phagocytosis activity of the peritoneal macrophages and delayed type hypersensitivity in mice bearing S180 were studied. The results showd that the ablong polysaccharide promoted the phagocytosis activity of the peritoneal macrophages and delayed-type hypersensitivity in mice bearing S180. It was suggested that the polysaccharide inhibited tumor growth maybe by activating the macrophage and T cells. PMID- 12575052 TI - [Orthogonal experiments on extract-concentrate-spray drier related technological factors for shuguanwenjing granule]. AB - By using orthogonal tests, with the ratio of dry powder abtained from crude drugs and the Icariin content of powder as criteria, the correlation among the extract concentrate-spray drier technology for Shuguanwenjing Granule and the main technological factors such as time and frequency of decoction, volume of water, relative density of extract, the in-temperature and out-temperature of spray drier, etc. was studied. PMID- 12575053 TI - [The effects of fertilization on yield and root diameter of Codonopsis pilosula var. modesla]. AB - Field trials were conducted using three factors regression design of quadratic orthogonal rotation. The result indicated that under the condition of cultivation, yield and average root diameter of Codonopsis Pilosula var. modesla (Nannf) L.T. Shen showed a positive correlation with N fertilizer application on high fertilization level; The effect of major three fertilizer on yield and average diameter of root was K > P > N. In the course of selective optimization yield which is higher than 3750 kg/hm2 (fresh weight) and average diameter of root which is more than 1.5 cm, the index of fertilizer application for N, P and K pure nutrient is 155 kg/hm2, 250 kg/hm2 (P2O5) and 60 kg/hm2 (K2O) respectively. The ration of N:P:K is 1:1.6:0.4. PMID- 12575054 TI - [The comparison of contents of uracil, xanthine and hypoxanthine in five species of leech]. AB - Three constituents(uracil, xanthine, hypoxanthine) were isolated from five species of leech and determined by reversed phase HPLC. The column employed was Shim-pack CLC-ODS C18(150 mm x 6 mm). The mobile phase was 0.05 mol/l ammonium phophate dibasic solution(pH = 8.4). The flow rate was 0.8 ml/min and detection was effected at 254 nm. This method is accurate, rapid and reproducible. Analytical data for five species and Whitmania pigra samples from different places were given. PMID- 12575055 TI - [Histological identification of Chinese drug zhiqiao & zhishi]. AB - After growing area investigating which covered 10 Provinces (or Cities) of China, we gathered 11 types Zhiqiao and 6 types Zhishi which consistently came from 12 species (or varieties). Histological studies on them have been taken. microscopical characters including transverse section and powder were described. These studies will provide scientific basis for the identification of Zhiqiao and Zhishi. PMID- 12575056 TI - [The origin of Chinese animal drug "hu-di-long" (Shanghai pheretima)]. AB - Results of identification showed that the origin of the Chinese animal drug "Hu Di-Long" is the dried body of Pheretima vulgaris Chen, P. guillelmi (Michaelsen) or P. pectinifera Michaelsen. The results have been specified in the Pharmacopoeia of the People's Republic of China (1995). PMID- 12575057 TI - [Isolation and identification of oleanonic acid from Patrinia scabiosaefolia]. AB - Oleanonic acid has been isolated from the acetone extracts of the roots and rhizomes of Patrinia scabiosaefolia Fish. ex. Link. Its structure was identified by the compare of its spectra data with that of Oleanonic acid. It was found in the patrinia for the first time. PMID- 12575058 TI - [GC-MS analysis of volatile oil of herba Pogostemonis collected from Gaoyao county]. AB - The chemical constituents of the volatile oil of the stems and leaves of Pogostemon cablin collected from Gaoyao county, Guangdong province, have been studied by means of GC-MS. Sixty four compounds have been identified. The main constituents are pogostone (30.99% in stems, 21.31% in leaves, the same below), patchouli alcohol (10.26%, 37.53%), transcaryophyllene (4.92%, 6.75%), alpha guaiene (2.27%, 6.18%) and seychellene (1.56%, 1.99%), etc. PMID- 12575059 TI - [Studies on chemical constituents of Lysimachia pentapetala]. AB - From the ethanol extract of Lysimachia pentapetala, six compounds have been isolated. These compounds were identified as triacontanol, beta-sitosterol, glycerol monopalmitate, succinic acid, flavonol glycosides and hitrite upon the date of mp, IR, MS and 1HNMR tests. PMID- 12575060 TI - [Analysis of chemical constituents in the essential oil from Evodia fargesii by GC/MS]. AB - In the essential oil from Evodia fargesii Dode, 33 constituents were identified by means of GC-MS-DS. The content of these compounds are 91.0% in the essential oil of this plant. The main compounds are eighteen terpenoids. PMID- 12575062 TI - [Influence of icariin on cell membrane of highly metastatic human lung tumor cell line]. AB - In this study, flow cytometry (FCM) and fluorescent probe technique were used to explore the influence of icariin (ICA) on cell membrane of PG. The data suggested that ICA could inhance membrane fluidity of PG and increase the expression of membrane HLA-ABC antigen. This could be one of the anticancer mechanisms of ICA. PMID- 12575061 TI - [Experimental studies of Chinese taihe chicken on effects of "Qi" reinforcing and "Yin" nourishing]. AB - The "Qi" deficiency and "Yin" deficiency rats were fed with Chinese Taihe Chicken or common Chicken. The results showed: Chinese Taihe Chicken and common Chicken could all improve in part the "Qi" deficiency of rats caused by forced swimming, furthermore, the effect of Chinese Taihe Chicken was better than that of common Chicken. Chinese Taihe Chicken could obviously reduce the activity of adenosine triphosphatase of kidney tissue of "Yin" deficiency rats caused by injecting thyroxine. It suggested that Chinese Taihe Chicken could nourish the "Yin". PMID- 12575063 TI - [Effects of places and stages on the contents of ganoderic acid B from Ganoderma lucidum]. AB - The paper reports the assay of ganoderic acid B from Ganoderma lucidum in different places and stages by means of HPLC. The results show that contents of ganoderic acid B from different cultured places have differnces, different stages have effect on the amount too. PMID- 12575064 TI - [Analysis of hemogram and parameters of biochemistry from sika blood]. AB - The hemogram and parameters of biochemistry were determined in eight male sika. The results showed that the red blood cell count was 125 x 10(8)/L, the mean corpuscular of hemoglobin was 38.7 pg and the mean corpuscular hemoglobin concentration was 970 g/L. It is suggested that there are rich hemoiderin and oxygen in sika blood. Moreover, the seven kinds of enzyme observed in the study were significantly higher than the human normal values. Serum levels of creatine phosphate kinase (CPK), hydroxybutyrate dehydrogenase (a-HBD) and CK-MB were 327 U/L, 639 U/L and 772 U/L respectively and which were seven to ten times higher than human normal values. It is indicated the activity of enzyme should be protected when the sika blood was used as raw material of medicine. PMID- 12575065 TI - [Chemical pattern recognition of traditional Chinese medicine xixin (I)]. AB - In this paper, three species of traditional Chinese medicine Xixin (Asarum heterotropoides Fr. var. mandshuricum (Maxim.) kitag, Asarum sieboldii Miq. var. seoulense Nakai and Asarum sieboldii Miq.) were identified by means of chemical pattern recognition, with 26 samples as training set and 19 samples as test set. The numerical characteristic features for identification were obtained from GC-MS analysis of volatile oils, all of obtained data were treated with PRIMA (Pattern Recognition by Independent Multivariate Analysis), thus computerized classification of Xixin samples was accomplished. The results agree with those from pharmacognosy. As a new method, chemical pattern recognition is especially suitable for identification of a large number of traditional Chinese medicine samples. PMID- 12575066 TI - [Pharmacognostical identification of Artemisia argyi and Artemisia indica]. AB - This paper reported the identification of Artemisia argyi and A. indica on their macroscopical and microscopical chracteristics, TLC and UV spectra. It provides a basis for their differentication, exploitage and utilization. PMID- 12575067 TI - [Chemical constituents of the essential oil from the fruits of Lindera glauca and its antifungal activities]. AB - The chemical constituents of the two essential oils extracted from the fruits of Lindera glauca by hydrodistillation and petroleum ether have been studied by means of GC-MS. Among which, 32 kinds of constituents were identified in distilling oil and 22 kinds in solvent oil. The main components in distilling oil were n-carpric acid (25.39%), germacrene A (10.71%), n-dodecanole acid (10.08%), epishyobunol acetate (7.29%) and caryophyllene oxide (5.44%), and in solvent oil were camphene (17.55%), 3,6,6-trimethyl-2-norpinene (16.85%), capric acid, ethyl ester (13.61%), eucalyptol (8.10%), and alpha-cis-ocimene (7.38%), In vitro the distilling oil exhibited more manifest antifungal properties than the solvent oil with MIC between 0.03-0.5 ml/L for pathogenic fungi species and 1.0-1.5 ml/L for moulds. Almost having not contained the sesquiterpenoids and their derivatives in the solvent oil maybe were a reason of poor inhibitory. PMID- 12575068 TI - [Effects of extract from Pegasus laternarius on the weight of immune organs and the antistress in mice]. AB - Extract from Pegasus laternarius Cuvier (PLE) could increase the splenic and thymic weight in immunity-suppressed mice induced by hydrocortisone or cyclophosphamide and could increase the thymic weight in senile mice. PLE could prolonged the survival time of anoxic mice under normal pressure or chemically anoxic mice poisoned by potassium cyanide. It could also prolong the survival time of mice swimming in ice-water and enhance the tolerance of mice without food to eat and water to drink. PMID- 12575069 TI - [Scavenging effects of Astragalus and Gynostemma pentaphyllum with its product on O2-. and .OH]. AB - The scavenging effects of Astragalus and Gynostemma pentaphyllum with its products on O2-. (produced by the autoxidation of pyrogallol) and .OH (generated from the Fenton reaction) were investigated by electrochemical method. The results showed that they could scavenge the active oxygen free radicals effectively, however, their scavenge ability was different for O2-. and .OH. The inhibitory effect of Ginsenoside Rb1 and Astragaloside IV on O2-. and .OH was the most strongest. Their IC50 values were calculated respectively. The scavenging ability of Astragalus samples on O2-. was in the order of the following: astragaloside IV > crude radix astragali > mongolia radix astragali > hebei radix astragali > radix astragali > radix astragali mixfry with honey; and the scavenging effect on .OH was: astragaloside IV > hebei radix astragali > radix astragali mix-fry with honey > radix astragali > crude radix astragali > mongolia radix astragali. The scavenging activity of gynostemma samples on O2-. was: ginsenoside Rb1 > gynostemma jujube tea > tabellae Jiaogulanosidi > anguo market gynostemma > hebei qingxian gynostemma; while on .OH, the inhibitory ability was: ginsenoside Rb1 > gynostemma jujube tea > anguo market gynostemma > hebei qingxian gynostemma > tabellae Jiaogulanosidi. PMID- 12575070 TI - [A fluorimetric study of the interactions of hydrochloridum berberini with vinylpyrrolidone and beta-cyclodextrin]. AB - The interactions of hydrochloridum berberini with 1-vinyl-2-pyrrolidone and beta cyclodextrin in aqueous solution were studied by spectrofluorimetric method. The spectral changes were quantitatively investigated. Results showed that the complexation with beta-cyclodextrin resulted in an enhancement of the fluorescence of hydrochloridum berberini whereas 1-vinyl-2-pyrrolidone caused a quenching of the fluorescence. It had been supposed 1:1 complex formation. Specifically, formation constants and the thermodynamic parameters at different temperatures were calculated. PMID- 12575071 TI - [Study on cultivation of Dendrobium flexicaule by habitat imitation]. AB - Dendrobium drug, a rare medicinal herb, reproduces differcut for growth environment to lead rare resources. By studying of cultivation for many many years in Fuliu mountain area, authors had found out the cultivated method simulating the habitat of wild Dendrobium flexicaule. The results could supply a basis for its development and utilization. PMID- 12575072 TI - [Extraction and purification of acidic mucopolysaccharide from Holothuria atra]. AB - Mucopolyaccharide with molecular weight of 10253 Da was extracted and purified from fresh Holothuria atra Jaeger by means of enzymic and alkaline hydrolysis, potassium acetate and ethanol fractional precipitation. It was tested to be purified ingredient with agarose electrophoresis. The percentage content of galactosamine, glucuronic acid, fucose and sulfate in the mucopolysaccharide was 16.12%, 17.88%, 11.66% and 23.52% respectively. PMID- 12575073 TI - [Chemical constituents of Scolopendra negrocapitis]. AB - The chemical constituent of Scolopendra negrocapitis Zhang et Wang were systematically analyzed and compared with S. subspinipes mutilans L. Koch. The results showed the lipid content was 18.7%, protein was 63.4% and total amino acid was 11.9%; S. negrocapitis contained the same twelve fat acids (the content of unsaturated fat acid was nearly 64% in its fat acids), twenty one amino acids and twelve trace elements as S. subspinipes mutilans had. The protein of S. negrocapitis showed forteen bands as determined by polyacrylamide gel electrophoresis. S. negrocapitis and S. subspinipes mutilans have the same main chemical composition. The contents of compositions are different. S. negrocapitis is worth as the nature resources of medicinal centepede. PMID- 12575074 TI - [Pharmacognostical studies on the roots of Euphorbia soongarica]. AB - Morphological and microscopical characters of the roots of Euphorbia soongarica Boiss. used as one of the origins of the Chinese medicine Daji in Xinjiang Autonomous Region of China were described and illustrated with line drawings. TLC and HPLC analysis of the above drug was also undertaken. These studies provide referencial informations for clinics, quality control, development and identification of this crude drug. PMID- 12575075 TI - [GC-MS analysis of volatile oil of Herba Pogostemonis collected from Leizhou county]. AB - The chemical constituents of the volatile oil of the stems and leaves of Pogostemon cablin collected from Leizhou county have been analysed by means of GC MS. The main constituents are patchouli alcohol, delta-guaiene, alpha-guaiene, seychellene, alpha-patchoulene, aciphyllene, trans-caryophyllene. PMID- 12575076 TI - [Determination of B(a)p contents of pine tar and its extractives]. AB - In this paper, the B(a)P contents of pine tar and its extractives A, B were determined by PC-Scanning method. After combining extraction by caffeine and formic acid with PC-Scanning method, the B(a)P contents were obtained. The results showed the B(a)P contents of pine tar and its extractives A, B were 6171.3 ppb, 2285.2 ppb, 129.6 ppb, among them, the B(a)P content of B was the lowest. PMID- 12575078 TI - [The regulation of Lycium barbarum on apoptosis of rat spleen in vitro]. AB - The regulation of Lycium barbarum (LB) on apoptosis of rat spleen induced by hydrocortisone (HYD) was studied in this paper. The results indicated LB could inhibit the apoptosis induced by HYD and the inhibition is dose dependent. PMID- 12575077 TI - [Effects of pure and crude Lycium barbarum polysaccharides on immunopharmacology]. AB - Effects of pure Lycium barbarum polysaccharides (LBP-X) on immunological activity were compared with crude LBP. The pure LBP-X were divided into different doses, lower doses (5-20 mg/kg.d) of pure LBP-X showed a remarkable effect on immunological enhancement. Especially, LBP-X 10 mg/kg.d had a highly significance difference compared with crude LBP on immune indices in mice. Lycium barbarum polysaccharides have the best pharmacological effect when it was the best dose. PMID- 12575080 TI - [Study on the inclusion effect of menthol with beta-cyclodextrin]. AB - Indusion effect of beta-CD with menthol was studied by three methods of preparation. Determination of physical constants and quantitative analysis proved that the inclusion compounds of beta-CD with menthol were formed, which was found to be 1:1 and 2:3. The results of thermal analysis demmonstrated that the gasifiation temperature of the inclusion compounds was by far higher than that of the free menthol. It was considered that the inclusion compound must be disconnected to be free, and then it can be determined by gas chromatography in the same condition of the free menthol. PMID- 12575079 TI - [Studies on the antitumour effect of Alocasia macrorrhiza]. AB - Models of transplanted tumour in mice and human cancer enograft in nude mice were used to evaluate the antitumour effect of water extract of Alocasia macrorrhiza. Results showed that the inhibitory rate against S180 in mice was 29.38%, and the inhibitory rate against transplantable humman gastroadenitis in nude mice was 51.72%. No antitumour effect was shown against ECA in mice. PMID- 12575081 TI - [The morphological variation in species of Anemarrhena asphodeloides]. AB - In this paper, morphological studies modified the description in some literatures. The features of Anemarrhena asphodeloides Bunge growing in different districts vary. Carefully observation under Scanning Electron Microscope (SEM) showed that surface characters of leaves at the same developing stage, which were collected from cultivated plants formerly growing in different districts, vary greatly but regularly. It was found that the morphology of pollen grains were similar. PMID- 12575082 TI - [Pharmacognostical identification of Iodes seuinii root, an adulterant of radix Stephaniae tetrandrae]. AB - In this aricle, the authors identified Iodes sequinii on Chinese medicinal material discriptions, microscopic characters, TLC and LV. This proved the evidences for Radix Stephaniae Tetrandrae medicinal material. PMID- 12575083 TI - [Pharmacognostical identification of one Rhizoma Drynariae habitually used species from Yunnan]. AB - The species recorded on the "Rhizoma Drynariae" item of Chinese Pharmacopeia is the dry rhizoma of Drynaria fortunei from Polypodiacae. But in Yunnan province, many plants (including D. delavayi) are habitually used as Rhizoma Drynariae. In this article the authors identified D. delavayi on Chinese medinal materials discriptions, microscopic characters and TLC. This proved evidences for the utilization and identification of D. delavayi in some extent. PMID- 12575084 TI - [Assay of three hydrolyzable tannins in Fructus Chebulae from different habitats by RP-HPLC]. AB - Three hydrolyzable tannins chebulinic acid (I), chebulagic acid(II) and 1,3, 6 tri-O-galloyl-beta-D-glucose (III) in Fructus Chebulae from different habitats were determined by RP-HPLC method. The contents of I and II were obviously interrelated with the variety and characteristics of Fructus Chebulae. It's suitable to use I and II as indexes in quality evaluation of the crude drug of Fructus Chebulae. PMID- 12575085 TI - [Analysis of chemical constituents in the essential oil from roots of Litsea rotundifolia var. oblongifolia by GC/MS]. AB - In this paper, 37 constituents in the essential oil from roots of Litsea rotundifolia var. oblongifolia were identified by means of GC-MS-DS. The content of these compounds are 86.37% in the essential oil of this plant, terpenoids is the main compound. PMID- 12575086 TI - [Effect of raw radish on pharmacological action of ginseng]. AB - Ginseng decoction can increase the memory-improvement, weary-resistance, antihypoxia, sedation in mice, and increase the weight of spleen and accessory sexual gland, but reduce the weight of thymus. Raw radish can decrease these pharmacological effects in varying degrees, except the sedation. PMID- 12575087 TI - [Effect of natrahagin, a Chinese cobra venom proteinase on hemorrheology in rat]. AB - The effects of natrahagin, a Chinese cobra venom proteinase on hemorrheology and plasma fibrinogen level in rat was investigated. The results showed that intravenous injection of natrahagin (0.025-0.1 mg/kg) dose-dependently decreased the whole blood viscosity and the plasma viscosity (P < 0.05 or P < 0.01) in rat, but caused no significant effect on hematocrit (P > 0.05). Natrahagin also decreased the plasma fibrinogen level (P < 0.05 or P < 0.01) of rat in a dose dependent manner at these doses. PMID- 12575088 TI - [Pharmacodynamic effects of the extracts from Orobanche cumana]. AB - Pharmacodynamic effects of the extracts from Orobanche cumana were studied for the first time. The result showed that it has the effects of antifatigue, facilitating immune function and an andrin-like action. It is consided that Orobanche cumana can be developed as a new medicinal resource plant. PMID- 12575089 TI - [Antitumor effect of the extract of birch bark and its influence to the immune function]. AB - OBJECTIVE: To study the antitumor effect of extract of birch bark (EBB) and its influence to the immune function of the tumor-bearing mice. METHOD: EBB was administered orally and the tumor inhibiting effects were observed in tumor bearing mice. Experiment of the immune function are to examine thymocytes (spleen) index, reaction of spleen to ConA and activity of NK cell. RESULTS: EBB showed antitumor effect in vivo to B16, S180 and Lewis lung cancer in mice. EBB can significantly prolong survival period of bearing tumor mice. EBB can enhance thymocytes and spleen index in C57 BL/6 mice. EBB can enhance splenic lymphocytes translate and activit of NK cell (P < 0.05). CONCLUSION: EBB have antitumor effect and enhance immune function in bearing tumor mice. PMID- 12575090 TI - [Study on the pharmacologic effect of propolis]. AB - Pharmacologic effects of propolis were investigated in this article. The results suggested that propolis has pharmacologic functions in many aspects. It is a new type medicine derived from animal in pharmacology and food both. To small mouse, propolis showed the functions of anti-fatigue and endurance to lack of oxygen. To high-blood-lipid-model mouse, it prevented increase of blood-mucus and blood lipid(TC, TG, LDL-C) (P < 0.05-0.01), but there were insignificant changes to red blood-cell proportion (HCT) and high-density lipoprotein chelesteral (HDC-C). To small immunosuppessive-model mouse, propolis could strengthen macrophagocyte phagocytosis in the abdominal-cavity (P < 0.05), and increase the thymus-index (P < 0.05), but there were insignificant changes to the spleen-index. The LD50 > 7500 mg/kg to small mouse. PMID- 12575091 TI - [Determination of resibufogenin in xinli pills by RP-HPLC]. AB - A method has been developed for the quantitative determination of Resibufogenin in Xinli pills by RP-HPLC. The stainless steel column(200 x 4.6 mm) was packed with ODS-3 (particle size 5 microns). The mobile phase was a mixture of methanol water (60:40). The flow rate was 1 ml/min. The column temperature was 30 degrees C. The detection wavelength was 299 nm. The retention time for recibufogenin was 5.6 min. The average recovery for resibufogenin was 99.9%. The linear calibration curve for resibufogenin was obtained in the range 53.0-159.0 micrograms with the correlation coefficient 0.9998. PMID- 12575092 TI - [Cuttage of Santalum album]. AB - The effects of cuttage times, miaternal plant ages, hormones and mediums on the taking root of a cutting were studied in 1991-1996. The results showed that the sprouts of germinating and growing 20-30 days from the cut back of maternal plant as cuttings, the rate of the taking root get to about 70%; the suitable cuttage time was in June to August; the proper medium was river sands, but the effects of hormones were not obvious. PMID- 12575093 TI - [Pharmacognostical studies on the roots of Euphorbia prolifera]. AB - Morphological and histological characters of the roots of Euphorbia prolifera Buch.-Han., a traditional Chinese medicine, were described and illustrated with line drawings. TLC analysis of the above drug was also undertaken. These studies provide referencial informations for clinics, quality control, development and identification of this crude drug. PMID- 12575094 TI - [GC-MS analysis and inhibitory activity of the essential oil extracted from the leaves of Lindera communis]. AB - The essential oil isolated from the dried leaves of Lindera communis was analyzed by means of gas chromatography-mass(GC-MS) technique, the structures of 23 chemical components were identified from it in total, among these, (-) spathulenol(relative content 22.50%), endo-1,3,3-trimethyl-2-norbornanol, acetate (10.06%), caryophyllene oxide (6.74%) and germacrene B(6.71%) were the main constituents. And also the oil in vitro exhibited manifest antifungal and antibacterial activities against 4 fungi pathogen species and 4 bacterial species with MIC between 0.08-0.8 ml/l. PMID- 12575095 TI - [Isolation of resisting thrombus and arteriosclerosis compounds in leaves of Salix matsudana]. AB - In this paper, three compounds were isolated and identified from the leaves of Salix matsudana. They are apigenin-7-0-beta-D-glucopyranside(I), luteolin-7-0 beta-D-glucopyranside(II), compound III. Compound I and II are isolated firstly from Salix spp., compound III is found firstly in the world. Furthermore, study on effect of arachidonic acid metabolisin in rat platelets by them with radio chromatography found that they can significantly inhibit the production of 12 HETE(12-hydroxy-5,8, 10,14-eicosatetraenoic acid), which can induce allergy and arteriosclerosis. The production of apigenin-7-0-beta-D-glucopyranside being hydrolyzed was apigenin, it can inhibit TXB2(thromoxane B2) which can induce platelet aggregation. PMID- 12575096 TI - [Studies on the supercritical-CO2 fluid extraction and quality evaluation of ginger oils]. AB - Essential oils from the dried rhizomes of Zingiber officinale were extracted by supercritical-CO2 fluid(SFE-CO2) and traditional water still distillation (WSD) methods, and the SFE extraction technology and quality (GC-MS, etc.) of the oils were studied. The results showed that the SFE-CO2 contains 49 constituents, such as 6-paradol, etc, the ginger peppery component of them gets to 22.90%. PMID- 12575097 TI - [Experimental study on anti-oxygen and promoting intelligence development of Codonopsis lanceolata in old mice]. AB - The water labyrinth test and determining lipid peroxide (LPO) contents were used to assay the effects of Codonopsis lanceolata on promoting intelligence development and lipid peroxide. The results showed that latent period of old mice in water labyrinth was statistically shorten and the number of mistake of old mice was visibly reduced by drinking extraction of Codonopsis lanceolate 4 g/kg and 8 g/kg. These indicated that mice drinking extraction of Codonopsis lanceloata learned faster that those of drinking water. The contents of LPO in red blood cell and brain of mice and rate drinking extraction of Codonopsis lanceolata were significantly reduced, the activities of superoxide dismutase (SOD) in serum of rat drinking extraction of Codonopsis lanceolata were visibly increased. These presents that the extraction of Codonopsis lanceolata has the role of promoting learning and memory of old animal, its mechanism might be increasing SOD activity and decreasing LPO level. PMID- 12575098 TI - [Effect of Callicarpa herbs on antioxidation in vitro]. AB - The concentration of half inhibition (IC50) of Callicarpa herbs, including Callicarpa kochiana(Ck), C. bodinieri (Cb) and its fruit (Cbf), C. japonica, C. macrophrlla, C. cathaarana, C. giraldi(Cg) on lipid peroxidation (LPO) of tissus homogenate incubated in 37 +/- 0.5 degrees C were tested. The result showed that all but Cg could inhibit lipid peroxidation. The IC50 of Cbf was minimum. IC50 of Cb, Cbf and Ck inhibiting red blood cell (RBC) LPO and hemolysis caused by hydroxyl free radical (.OH), inhibiting brain homogenate LPO induced by .OH and inhibiting or clean .OH formation by system of H2O2 and FeSO4 were also tested. It was found the IC50 were Cbf > Cb > Ck. These results suggested that the most strong inhibitor to automatic LPO reaction was Cbf, but to LPO caused by .OH was Ck. PMID- 12575099 TI - [Inhibiting effects of the ethanol-soluble extracts from the dried rhizoma of Garcinia oblougififolia on hepatitis B virus in vitro]. AB - Comparing with Ara-Amp, the effects of the ethano-soluble extracts from the dried rhizoma of Garcinia oblougififolia Champ (GOC) on HBeAg and HBsAg expression in 2.2.15 cells, which came from transfected HepG2 cells with coloned HBV DNA, were studied. The results showed that the extracts of GOC had a marked inhibition effects on HBeAg adn HBsAg which expressed by 2.2.15 cells, the effective concentration was range from 195 to 780 micrograms/ml. PMID- 12575100 TI - [Karyotype analysis of Carpesium cernuum]. AB - The paper reported the chromosome number, karyotype and chromosome volume of Carpesium cernuum. The normal diploid was 2n = 40, karyotype formula based on Levan's publication was k(2n) = 40 = 24 m + 12 sm + 2 st + 2T. According to the method of S. R. Kuo, the chromosome relative length was 2n = 40 = 8L + 6M2 + 16M1 + 10S, which belong to "2B" type according to the Stebbins' karyotype classification. The AS.K% was 62.35% from Arano's method. The total volume of chromosome groups was 13.76 microns 3. PMID- 12575101 TI - [Biological characters of the different varieties for Luffa cylindrica]. AB - The yield of Luffa cylindrica cultivated in Taiyuan of Shanxi Province, which introduced from Zhejiang Province, was obviously high than that of local variety. The paper reported the biological characters of the two varieties. PMID- 12575102 TI - [Analysis of the vestiges of farm chemicals for Panax notoginseng]. AB - The vestiges of farm chemicals in Panax notoginseng plants and medicinal materials from main production areas in Wenshan District was analyzed in this study. The result showed that the samples from the places contained hexachloro cyclohexane and DDT. And the vestiges of hexachloro-cyclohexane was about 0.016 0.101 mg/kg, that of DDT was about 0.15-0.143 mg/kg. The content of Cu, Zn, Pb, Cd was lower than national standard of P. R. C. PMID- 12575103 TI - [Pharmacognostical studies on radix Glehniae (Glehnia littoralis)]. AB - OBJECT: To identify the roots of Glehnia littoralis Fr. Schmidt ex Miq., and compare the chemical constituents of the root skin and the roots with no skin. METHODS: The roots were identified by morphological and microscopic identification and TLC. RESULTS: The characteristics of the secretory canal, ray and starch grain can be used to identify the histology and powder of the roots. The chemical constituents of the root skin and the roots with no skin are similar. CONCLUSION: The characteristics of the morphology, histology and powder can be used to identify the roots of Glehnia littoralis (Radix Glehniae). PMID- 12575104 TI - [Isolation and identification of a saponine from Patrinia scabiosaefolia]. AB - A saponine compound was isolated from the acetone extract of the roots and rhizomes of Patrinia scabiosaefolia Fish. ex Link. Its structure was identified by combination of chemical reaction and spectrum analysis as 3-O-alpha-L rhamnopyranosyl-(1-2)-beta-D-xylopyranosyl oleanolic acid (giganteaside D), and part of its 13C-NMR data was revised by the 2D-NMR. The compound was found in the Patrinia for the first time. PMID- 12575105 TI - [Chemical constituents of Girardinia palmata ssp ciliata]. AB - Thirteen constituents were identified from Girardinia palmata ssp ciliata C. J. Chen by using GC-MS, which amounted to 84.5% of the total peaks, the main constituent were linoleic acid (52.01%) and dimethyl fumarate (10.05%). The dissociated amino acids and KNO(3) in the sample amounted to 43.30 mg/100 g and 3.0%. Five compounds were isolated and elucidated as octadecane, hexadecylic acid, beta-sitosterol, p-coumaric acid and potasium nitrate. PMID- 12575106 TI - [Synergy and attenuation of cyclophosphamidum (CTX) activities by abalone polysaccharide]. AB - Abalone polysaccharide (AP) was extracted from Haliotis discushannai Ino. The combined effects of AP and CTX against tumor-bearing mice were investigated. The results showed that AP could markedly promote the inhibition effects of CTX on S180 and HepA tumor, enhance protection against leukocytopenia, decrease of spleen and thymus weight and hemolysin in serum, and bone marrow supression induced by CTX. PMID- 12575107 TI - [Comparison of effects of Herba Pogostmonis from gaoyao and wuchuan on digestive system]. AB - The effects of the three extracts (decoction, oil-free decoction and volatile oil) of Herba Pogostmonis from Gaoyao and Wuchuan on digestive system were compared. The results demonstrated that the decotion and oil-free decoction from Gaoyao and Wuchuan could depressed gastric evacuation in mice. All the three extracts from Gaoyao and Wuchuan could inhibit the ACh-as well as BaCl2-induced spasmodic and automatic contraction of isolated rabbit intestine. The decoction and oil-free decoction from either Gaoyao or Wuchuan inhibited the normal intestinal propulsion. They also increased gastric secretion of acid and activity of pepsin and amylase. These suggest that both of Herba pogostmonis relief gastrointestinal spasm and improve the digestion. The total effect of Herba pogostmonis from Gaoyao was more potent that from Wuchuan. PMID- 12575108 TI - [Study on the inclusion of oridonin-beta-cyclodextrin]. AB - The inclusion compound of Oridonin-beta-cyclodextrin was prepared by the liquid phase method in aqueous solution. And it was confirmed by thin layer chromatography, differential thermal analysis and specific rotation. The change (A) of ultraviolet spectrum was determined. The composition ratio of inclusion compound between Oridonin and beta-CD was obtained by the continuous variation method. PMID- 12575109 TI - [Effects of phytohormones on growth and content of depsides in Salvia miltiorrhiza suspension cells]. AB - This paper deals with the effects of 2,4-D, BA and GA3 on the growth and content of two depsides (rosmarinic acid and lithospermic acid B) in suspension cells of Salvia miltiorrhiza. The results showed cell growth and rosmarinic acid content reached the maximum on the 12th day and lithospermic acid B content on the 16th day after incubating cells in the subculture medium MS + 2, 4-D 1 mg/L + KT 0.1 mg/L. This cell line was a growth-product-associated. With the same concentration (1 mg/L), 2, 4-D stimulated the cell growth but prohibited the formation of lithospermic acid B; GA3 inhabited the cell growth but stimulated the formation of two depsides; The effects of BA is between 2, 4-D and GA3. The concentration optimum of phytohormones tested displayed 3 mg/L for BA and 1 mg/L for GA3. For the optimum time of adding BA and GA3 was in med-term (the 8th day) and early term (at the beginning) of culture period respectively. The synergiatic function of BA and GA3 on the depside formation was also showed that adding GA3 (1 mg/L) was favour for the formation of lithospermic acid B of the suspension cell cultured in the medium containing BA 3 mg/L. The suitable time of adding GA3 was the 6the day after incubating cells in the medium of MS + BA 3 mg/L. PMID- 12575110 TI - [Application of RAPD in the taxonomy of the genus Fritillaria]. AB - OBJECTIVE: To provide the molecular data for the right application of the Chinese medicine "Beimu". METHOD: Using the technology of RAPD, we studied the relationship among 12 samples of Beimu. RESULT: The total genomic DNA of all the samples are about 21 Kb in size. Among 20 primers used, the five primers can reatedly generate a certain specified amplified band type, 27 bands were recored from all amplified products and 25 polymorphic fragments were found in it. The size of amplified fragments is between 450 bp and 1904 bp. CONCLUSION: The similarity within species is higher than those between species. The relationship of Fritillaria anhuiensis and F. puqiensis is the farest, while F. thunbergii and F. puqiensis is the closest. PMID- 12575111 TI - [Pharmacognostical studies on the roots of Stellera chamaejasme and S. chamaejasme f. chrysantha]. AB - Morphological and histological characters of the roots of Stellera chamaejasme L. and S. chamaejasme L. f. chrysantha S. C. Huang, used as the origins of the Chinese medicine Langdu, were described and illustrated with line drawings. TLC analyses of the above two drugs were also undertaken. These studies provide referencial informations for the quality control, development and identification of these two drugs. PMID- 12575112 TI - [Morphological and commercial identification on semen torreyae]. AB - The morphological characters and commercial identification on Semen Torreyae has been reported. The results of identification of commercial samples indicated that Torreya grandis was mainly used; T. fargesii, T. yunnanensis and T. jackii were used in their producing areas. PMID- 12575113 TI - [Studies on the chemical constituents of Viscum coloratum]. AB - Four compounds were isolated from Viscum coloratum (Kom.) Nakai by means of both solvent extraction and chromatographic separation methods. These constituents were identified through spectral analysis of IR, 1H-NMR. 13C-NMR and MS. They are syringin, oleanolic acid, beta-acetylamyrin and homoeriodictyol-7-beta-D glucoside. It is the first separation from freshly collected Viscum colortum (Kom.) Nakai, which parasitizing on pear tree. PMID- 12575114 TI - [Study on lipids and other volatile constituents in Pheretima aspergillum]. AB - To study chemical constituents in Pheretima aspergillum, three kinds of fractions were obtained from this drug by soxhlet extraction with different solvents, and the chemical structures of thirty-six volatile components were identified by means of GC-MS. The eleven in ether fraction were all lipids and the relative content of non-saturated fatty acid was the highest(27.70%) such as oleic acid, linoleic acid, arachidonic acid and eicosatrienoic acid; There were eight lipids in acetone fraction (35.75%), which included one kind of nonsaturated fatty acid (linoleic acid); There were thirteen lipids in ethanol fraction (72.09%), which non-saturated fatty acid has never been detected. This study has determined the lipid composition in Pheretima aspergillum, especially non-saturated fatty acid, and afforded chemical base to cardio-cerebro-vascular therapy. PMID- 12575115 TI - [Study on the volatile oil of Nardostachys chinensis]. AB - The volatile oil of Nardostachys chinensis Batal has been studied by GC-MS, and 21 compounds have been isolated and identified. The total content of the components are 88.51%. Major compounds have been identified as Calarene (29.44%). delta 1(10)-Aristolenone-2 (16.57%) and Jatamansinol (8.80%). PMID- 12575116 TI - [Studies on chemical constituents of fat oil of Polygala tenuifolia]. AB - OBJECTIVE: To study the chemical constituents of fat oil from root of Polygala tenuifolia Willd. METHOD: The components were separated and identified by gas chromatograpy-mass spectrometry and elucidated on the standard mass spectral data and the percentages were determined by gas chromatographic area normalization method. RESULT: Eighteen constituents were separated and sixteen of them were identified. CONCLUSION: The major components of the fat oil were established first time as oleic acid, linoleic acid, palmitic acid, eicosenoic acid and stearic acid, which make up 87.0%, 7.31%, 3.27%, 1.07%, 0.91% of fat oil respectively. PMID- 12575117 TI - [Analysis of essential oil from Amomum villodum by extraction of supercritical CO2 fluid]. AB - By using the extraction technique of supercritical-CO2 fluid on Amomum villodum Lour, essential oil could be got and then be applied to GC-MS online computer retrieval analysis, after a study on essential oil both in isolated pot I and II, 24 and 71 components were identified respectively. PMID- 12575118 TI - [Inhibition of extracts of ampelopsis sinica roots on DHBVsAg in sera of ducklings]. AB - The ducklings, which are infected with Duck HBV to produce DHBVsAg in sera in advance, are treated with extracts from Ampelopsis sinica roots and some of them are treated with interferon alpha (INF) as positive control group. According the titer of DHBVsAg in Ducklings sera and the results of observing the tissues of the duckling's livers, it is demonstrated that the extracts of Ampelopsis sinica roots possess a certain action inhibiting DHBV and the action is enduring. PMID- 12575119 TI - [Study on the quality standard of changan oral solution]. AB - OBJECTIVE: To study the quality standard of Changan oral solution. METHOD: Thin layer chromatography was used for identification pig's bile plaster. TLC scanning was used for the determination of hyodeoxycholic acid. RESULTS: The TLC identification was highly specific and the spots was clear and concentrated. Linear regression for hyodeoxycholic acid was over the range of 3.50-12.25 micrograms. The average recovery of hyodeoxycholic acid was 100.5%, RSD was 3.2%. CONCLUSION: This standard was capable to effectively control the quality of Changan oral solution. PMID- 12575120 TI - [Herbological investigation on traditional Chinese medicine gegen]. AB - In this paper, herbological investigation was carried out into a large of Ben-Cao literature that concerned the origin, clinical use and other relevant aspects of Gegen, from which we can judge that Gegen used in ancient times match to that specified in the present Chinese Pharmacopoeia. PMID- 12575121 TI - [Biological effects of supplements on soil properties and the growth of Panax quinquefolium]. AB - This paper suggests that the physical and chemical characteristics of soil in American ginseng field were greatly improved after utilization of supplement on soil. Compared with the contrast, the soil gravity ratio decreased from 0.812 g/cm3 to 0.715 g/cm3, while the soil porosity increased from 69.36% to 73.03%. These improvements are benefit for the growth of Panax quinquefolium. Biological observations show that the fresh weight of ginseng root enhanced 53.8%, and the content of total saponins in root enhanced 0.5%-1% (get to 8.28%). PMID- 12575122 TI - [Concentration of four alkaloids in the aerial parts of Eomecon chionantha from different mounth in year]. AB - The concentration of four alkaloids (sanguinarine, chelerythrine, protopine and alpha-allocryptopine) in the aerial parts of snowpopy (Eomecon chionantha) collected from April to August in 1996 were determined by capillary electrophoretic method. The results show that the change of contents of alkaloids according to months is similar to that of the underground parts of snowpopy, i.e. the contents of sanguinarine and chelerythrine wrer lower in June than other months; the contents of chelerythrine were higher than that of sanguinarine and alpha-allocryptopine were higher than protopine. PMID- 12575123 TI - [Morphological and microscopic identification of herba artemisiae scopariae and its adulterants]. AB - Pharmacognostical studies of Herba Artemisiae Scopariae and its adulterants were compared on morphological and microscopic characteristics. The detailed characteristics of powder microscopic identification were described. PMID- 12575124 TI - [Pharmacognostical studies on the stem and leaf of Mussaenda pubescens]. AB - Morphological and microscopic chracteristics of Mussaenda pubescens Ait. f, a traditional Chinese medicine were described and illustrated with line drawings. TLC and UV analysis of the medicinal material were also undertaken. These studies provide referential information for identification and development of this medicinal material. PMID- 12575125 TI - [Determination of organic acids of tetraploid Isatis indigotica with high performance capillary electrophoresis]. AB - The content of organic acids, being antiendotoxic activities in vitro, of tetraploid Isatis indigotica Fort. were determined with high performance capillary electrophoresis. The results showed that the contents of organic acids of tetraploid Isatis indigotica Fort. were obviously higher than that of diploid parent. Tetraploid Isatis indigotica Fort. was a new lines with better quality, which had notably value of exploiting and utilizing. PMID- 12575126 TI - [GC-MS analysis of volatile componments in root of Ficus hitra]. AB - The volatile components in root of Ficus hitra Vahl were analyzed by GC-MS, the results suggested that the main volatile components in root of Ficus hitra Vahl were Hexadecanoic acid, Oleic acid, Linolic acid and Ethyl Acetate; its ether extract contains Isopsoralen and Bergapten. Coumarins should be one of the effective sections in root of Ficus hitra Vahl. PMID- 12575127 TI - [Studies on antiplatelet aggregation and anticoagulant action of Curcuma phaeocaulis]. AB - OBJECTIVE: To study the effects of Curcuma phaeocaulis Valeton on platelet aggregation and anticoagulant action. METHODS: Antiplatelet aggregation and anticoagulant action of Curcuma phaeocaulis Valeton of mice is observed in platelet aggregation and coagulant time. RESULTS: The experimental results show that Curcuma phaeocaulis Valeton can inhibit platelet aggregation, prolong time of mice coagulation. CONCLUSION: Antiplatelet aggregation and anticoagulant action of processed products with vinegar is the most powerful. PMID- 12575128 TI - [Improving memory effects of eel oil capsule on memory obstruction of mice hurt by anisodine]. AB - The effects of eel oil capsule on memory obstruction of mice were observed in step-down test, step-through test, Y-maze test and Japanese labyrinth test. The results indicated that the eel oil capsule treating group with dosages of 0.234, 0.702 and 2.106 g/kg markedly enhance the acquirement, strengthing and reappearance of memory, having the functions of improving brain memory hurt by Anisodine. PMID- 12575129 TI - [Applying pharmaceutical effect indexes to screen out the preparation technology of xiaobanxia added fuling granules]. AB - After having adopted vomiting animal model created by filling stomach with cuprum sulfuricum and after having selected incubation period and vomiting index as our measuring and commenting indexes. We prepared decoction of Xiaobanxia added Fuling granules by applying orthogonal experimental method with four factors and three levels to proceed a preferable choice of extracting technology. The superior extracting technology became A2B3C2, namely: comminuting medicinal materials into coarse powder, for the first time, adding 9 times dosage of water and decocting for 45 minutes, for the second time, adding 5 times dosage of water and decocting for 30 minutes, condensing the filtrate to light paste (specific gravity 1.25-1.27, assaying at 25 centigrade), preparing decoction of Xiaobanxia added Fuling granules with a proportion as below:light paste:amylin:saccharose powder = 1:3:0.5. PMID- 12575130 TI - [A preliminary study on tissue culture of stem apex of Chrysanthemum morifolium]. AB - To solve the breed degeneration of Chrysanthemum the stem apex of Chrysanthemum morifolium used as explants were inoculated on MS medium with different pH and supplemented with different ratio of plant hormone. The results showed that when pH was between 5.8 and 6.4, the rate of callus were all over 80%. While explants inoculated on MS medium supplemented with NAA 0.2 mg/L and KT 0.1-0.2 mg/L, the rate of callus was noticeably higher than that of NAA 0.2 mg/L and 6-BA 1-3 mg/L. The preferable medium which roots could be induced was 1/2 MS + NAA 0.2 mg/L. PMID- 12575131 TI - [Comparison of total saponin content in three wild species of Gynostemma on Mount Emei]. AB - Analysed by TLC and spectrometry, the content of the total saponin in three wild species of Gynostemma on Mount Emei is found to be the highest in July and to be the lowest in the seedling stage. Among the three, the content of the total saponin in Gynostemma pentaphyllum is the highest, that in Gynostemma pubescens lower and that in Gynostemma longipes the lowest. PMID- 12575132 TI - [Studies on pharmacognosy of Echinacea purpurea]. AB - Pharmacognostical studies of Echinacea purpurea were carried out by botanical analysis, microscopic analysis and physicochemical analysis. The detalied pharmacognostical characteristics of Echinacea purpurea were described respectively. PMID- 12575133 TI - [Analysis of essential oil from Amomum tsaoko by extraction of supercritical CO2 fluid]. AB - By using the extraction technique of supercritical CO2 fluid on Amomum tsaoko Crevost et Lemaire, essential oil could be got and then be applied to GC-MS online computer retrieval analysis. After a study on essential oil both in isolated pot I and II, 65 and 93 components were identified respectively. PMID- 12575134 TI - [Chemical composition of essential oil in stems, leaves and flowers of Agastache rugosa]. AB - The chemical components and their relative contents of essential oil in different parts (stems, leaves and flowers) of Agastache rugosa have been analyzed by GC-MS technique, and 32 kinds of chemical structures in these three kinds of oils have been identified. Among which, it included 24 kinds in leaf-oil, 27 in stem-oil and 22 in flower-oil. The principal components in all these three kinds of oils were methylchavicol (60.01-88.43%), and other important components were d limonene, caryophyllene, hexadecanoic acid, linoleic acid, octahydro-7-methyl methylene-4-(1-methylethyl)-1H-cyclopenta [1, 3] cyclopropa [1, 2] benzene, etc. PMID- 12575135 TI - [The influence of external stimulation on content and quality of volatile oil in Lignun Santali albi]. AB - The authors analyzed the quality of Ligmum Santali Albi formed by the external stimulation of hormone and windburn by GC-MS-DS. The results showed that the content of volatile oil is 2.34% in the heart wood formed in 10 years tree age of Santalum album (SA) after 2 years stimulation continuously with a definite concentration of hormone, which is near to the 25 years tree age of SA in the same place. The GC-MS analysis showed that the content of santalol and other chemical components in volatile oil are similar to the 25 years tree age of SA. It is indicated that a definite concentration of hormone stimulated the SA may shorten the formation of the heart wood. The heart wood can be also formed by the broken branches after 2 years windburn, but its content of volatile oil is only 1/2 of the heart wood formed by hormone stimulation. PMID- 12575136 TI - [Study on primary speciation of seven metallic elements in pearl powder]. AB - We have determined seven metallic elements in various extract of pearl powder by FAAS. The result implies that the quantity of different element in water and acidic solvent system is different. It has supplied reference data for the further studies of speciation of metallic elements in natural product also. PMID- 12575137 TI - [Study of xixin, duzhong and mixture to the antioxidant system of D-galactose induced senile mice]. AB - D-galactose induced mice aging model were used in this experiment. This study investigated the effect of the herb on the activity of superoxide dismutase(SOD), glutahione peroxidase (GSH-Px) and NOS, in order to know the damages degree of body by free radical and determine the changes of above index after giving Duzhong, Xixin and their Mixture 15, 30 and 45 days, at the same time, the mechanism of their effect on antiaging was probed. The results were as follows: The activity of SOD, GSH-Px and NOS was increasing before middel age and decreasing after middelage(P < 0.01); Xixin and Duzhong could enhance the activity of SOD, GSH-Px and NOS; The time of effect for mixture and Duzhong to above index was short than Xixin's. The time of Mixture and Duzhong was 15 days, and Xixin was 30 days. PMID- 12575138 TI - [Study on erzhi pill effects on neuro-endocrinologic-immuno network of animal of deficiency of yin]. AB - OBJECTIVE: To investigate Erzhi Pill effect on Neuro-Endocrinologic-Immuno network of animal of deficiency of Yin. METHOD: In the experiment through the determination of hemotic cortisol, insulin, testosterone, beta-endorphin, cholesterol, weight of adrenal and thymus, ConA-induced, spleelymphocyte proliferation and activity of natural killer cells (NKC). The changes of the animals of deficiency of Yin and the effects of the Erzhi Pill were investigated. RESULT: It showed that rats of deficiency of Yin, their hemotic, cortisol, insulin, testosterone, beta-endorphin, weight of adrenal and thymus decreased, cholesterol increased and mouse of deficiency of Yin, their lymphocyte proliferation and activity of NKC dcreased, which might be modulated with Erzhi Pill. Erzhi Pill could improve hemotic cortisol, insulin, testosterone, beta endorphin, weight of adrenal and thymus, reduce hemotic cholesterol, improve lymphocyte proliferation and activity of NKC. CONCLUSION: This principle could improve animals of deficiency of Yin regulating Neuro-Endocrinologic-Immuno network and keeping homeostais. PMID- 12575139 TI - [Tissue culture and rapid propagation of Aloe arborescens]. AB - Tissue culture and rapid propagation of Aloe arborescens have been studied. The main results are as follows: the optimum medium for cluster shoots induction is MS + 6-BA 3.0 mg/L + NAA 0.2 mg/L; for cluster shoots propagation is MS + 6-BA 2.0 mg/L + NAA 0.2 mg/L; for roots induction is 1/2 MS + NAA 0.5 mg/L. The survival rate of the tube plantlets is 100% in the transplanting. Cutting down the cost of medium in large scale of production have been studied also. PMID- 12575140 TI - [Comparison of akebia saponin D contents in radix Dipsaci from different area]. AB - According to Akebia saponin D contents determined by HPLC, this article analyzed the quality of Radix Dipsaci grown in different area. The results indicated that the quality of Radix Dipsaci from the Muli and the Yanyuan of Sichuan province is superior to that of other province. PMID- 12575141 TI - [Analysis of Amomun villosum species and some adulterants of zingiberaceae by RAPD]. AB - Different Amomum villosum Lour. species and some their adulterants of Zingiberaceae were researched using RAPD. The PCR indicates favourable differentiation of the reaction. The Amomum villosum Lour. species have the similar DNA fingerprints while it is obviously differ from adulterants. There are 12.17 percent of primers which appear polymorphism. We can identify different species according to these dissimilarity. We also construct the tree of the molecular evolution through NJ software, the hereditary distance of the graph shows different relationships between Amomum villosum Lour. species and their adulterants. The result created by software is analogous the traditional methods. PMID- 12575142 TI - [Pharmacognostical studies on the roots of Euphorbia stracheyi]. AB - Morphological and histological characters of the roots of Euphorbia stracheyi Boiss., a traditional Chinese medicine, were described and illustrated with line drawings. TLC analysis of the above drug was also undertaken. These studies provide referencial information for clinics, quality control, development and identification of this crude drug. PMID- 12575143 TI - [Chemical study of Alpinia officinarum]. AB - Seven compounds were isolated from Alinia officinarum Hance and were identified as beta-sitoterol, 1,7-diphenyl-5-ol-3-heptone, 1-phenyl-7-(3'-methoxyl-4' hydroxyl) phenyl-5-ol-3-heptone, glandin, kaempferol-4'-methylether and 3,4 dihydroxylbenzoic acid by IR, 1HNMR, 13CNMR, FAB-MS and EA. Among these compounds, 3,4-dihydroxylbenzoic acid was the first time obtained from Alpinia officinarum Hance. Furthermore, 1-phenyl-7-(3'-methoxyl-4'-hydroxyl) phenyl-5-ol 3-heptone and a new compound 1,7-diphenyl-3,5-heptandiol-phenyl-7-(3'-methoxyl-4' hydroxyl) phenyl-3,5-heptaxdiol were obtained from 1,7-diphenyl-5-ol-3-heptone and 1-phenyl-7-(3'-methoxyl-4'-hydroxyl) phenyl-5-ol-3-heptone via chemical reductions. PMID- 12575144 TI - [Analysis on essential oil from Alpinia katsumada by extraction of supercritical CO2 fluid]. AB - By using the extraction technique of supercritical CO2 fluid on Alpinia katsumada Hayata, essential oil could be got and then be applied to GC-MS online computer retrieval analysis. After a stuy on essential oil both in isolation pot I and II, 95 peaks and 126 peaks were recorded respectively, and 81 and 120 components were identified respectively. PMID- 12575145 TI - [Protective effect of total paeony glycoside against cerebral ischemia reperfusion injury in mice]. AB - OBJECTIVE: To investigate actions of Total Paeony Glycoside (TPG) on cerebral ischemia-reperfusion model in mice. METHODS: The cerebral ischemia-reperfusion models induced by lagation of bilateral common carotid arteries incompletely were used. RESULTS: It was found that TPG could improve the learning and memory capacity of model mice in step down test. TPG significantly reduced the decrease of superoxide dismutase(SOD), inhibited the increase of nitric oxide(NO) and lessened the level of malondialdehyde(MDA) in model mice. 75 mg.kg-1 TPG also reduced the decrease of lactate dehydrogenase (LDH) in cerebrum remarkedly. CONCLUSION: It is suggested that TPG possess obvious protective effects against cerebral ischemia-reperfusion mice. PMID- 12575146 TI - [Study on the extraction of volatile oil in dry ginger and the preparation of its beta-cyclodextrin inclusion compound]. AB - By using orthogonal tests, the optimized extraction of volatile oil was practiced by distillation. The inclusion compound of the oil-beta-cyclodextrin inclusion compound was prepared by the liquid-phase method in aqueous solution. And it was confirmed by thin layer chromatogram and differential thermal analysis. PMID- 12575147 TI - [Study on the callus and cell culture of Hypericum perforatum]. AB - Callus were induced from the leaves and stems of Hypericum perforatum L. The cell suspension culture and plate culture were also investigated in this paper. The results showed that the medium of MS which contained 2,4-D 1 mg/L and NAA 1 mg/L was the best one for inducing callus. The plant growth regulator of 2,4-D was assential for callus and suspension culture. Factors such as the time of cell suspension culture, the ways of plating and cell density, which affects the plating efficency were studied. The results showed that plating at density of 5 x 10(3) cells/ml and with monocells seperated from the cells cultured 14-21 days, high plating efficiency could be obtained either by conditional plate culture or by nursing plate culture. PMID- 12575148 TI - [Influences of counterfeits on RAPD results of Semen Cuscutae]. AB - Compared the differences of the RAPD results between pure Semen Cuscutae and 12 commercial samples, it comes into the conclusion that the exiting other sample influences the RAPD results when its content is over 60% and the results is same as its content being under 60%. PMID- 12575149 TI - [Pharmacognostical studies on the roots of Euphorbia fischeriana]. AB - Morphological and histological characters of the roots of Euphorbia fischeriana Steud., used as one of the origins of traditional Chinese medicine Langdu, were described and illustrated with line drawings. TLC analysis of the above drug was also undertaken. These studies provide referencial informations for clinics, quality control, development and identification of this crude drug. PMID- 12575150 TI - [Identification of herba pogostemonis and its confused species Anisomeles indica]. AB - The article reports the morphological, histological and TLC identification for Herba Pogostemonis and its adulterant, Anisomeles indica, which appears recently. It provides foundation for identificating Herba Pogostemonis. PMID- 12575151 TI - [Electrophoresis identification of four kinds of seed-fruit Chinese drugs and its adulterants]. AB - OBJECTIVE: To identify four kinds of seed-fruit Chinese drugs and its adulterants by electrophoresis and to research the dyeing effect of Coomassie Brilliant Blue G-250. METHOD: Gel electrophoresis of soluble protein. RESULTS: The electrophoretograms of the four kinds of Chinese drugs and its adulterants are remarkably different. CONCLUSION: The electrophoretograms can be used to differentiate the four kinds of Chinese drugs and its adulterants. Coomassie Brilliant Blue G-250 can be used in the electrophoresis identification. PMID- 12575152 TI - [Studies on the supercritical CO2 fluid extraction and separation of psoralen, isopsoralen and fatty oils from Psoralea corylifolia]. AB - Opimum extraction conditions were studied by supercritical carbon dioxide fluid extraction technology and orthogonal tests. Crystal of psoralen can be achieved directly from the extractives. Magnified experiments were carried out. The chemical constituents of fatty oils were analyzed by GC-MS. The result shows that the optimum conditions are: extraction pressure 27 MPa, extraction temperature 70 degrees C; seperation pressure 9 MPa, seperation temperature 60 degrees C. Six components such as oleic acid, linoleic acid etc. were identified in the fatty oils. PMID- 12575153 TI - [Determination of naringin and synephrine in fructus aurantii from different habitats by HPLC]. AB - This paper reports the determination of Naringin and Synephrine in Fructus Aurantii from different habitats by HPLC. The result shows the highest content of Naringin in Citrus aurantium L. 'Xiangcheng' from genuine crude drug habitat Xingan city Jiangxi provice, the highest content of synephrine in Citrus aurantium L. 'Xiangcheng' from Yiyang city Jiangxi provice. Quality evaluation of genuine crude drug was studied preliminarily. PMID- 12575154 TI - [Study on the inhibitory activity, in vitro, of baicalein and baicalin against skin fungi and bacteria]. AB - In this paper, we concentrated in examining, in vitro, the antiseptic activity of the baicalein and baicalin upon the seventeen pathogenic skin fungal and sixteen skin bacterial strains, these two flavonic compounds were known principally as the biosubstances of a traditional Chinese medicinal plant: Scutellaria baicalensis. In agar media, the baicalein possessed potent specific activity against the pathogenic yeasts with MICs of 70-100 micrograms/ml; But in the same condition, no inhibitory effect was observed upon dermatophytes and filamentous imperfect fungi for baicalein, and upon all used strains for baicalin. According to the antibacterial test of baicalein, a high efficacy was achieved against certain causative specie of axillary and foot's odour such as Micrococcus sedentarius, Staphylococcus epidermidis, S. hominis and C. xerosis with a MICs inferior to 250 micrograms/ml. The good inhibitory activity of baicalein could be linked to the group hydroxyl (-OH) in position seven of the molecule. PMID- 12575155 TI - [Study on inhibitory actions of san huang yi gan capsule (SHYGC) on HBeAg with seropharmacological method]. AB - In this paper, the inhibitory actions of SHYGC on HBeAg in vitro were studied with the seropharmacological method. By Enzyme Linked Immuno Sorbent Assay (ELISA), it was found that the rabbit sera containing SHYGC have significant inhibitory effects on HBeAg, and they become stronger with the drug concentration in sera improved and the actions time prolonged, they decrease with the HBeAg concentration improved, and the effects of the sera containing hing-dose drugs equal those of Su Xiao Jing containing 250 micrograms/g effective chloric. The direct external inhibitory effects of SHYGC in original pharmaceutics are stronger than those of the sera containing drugs. It probably indicated that the active ingredients of SHYGC could not be digested and absorbed completely from gastrointestine, or were inactived by metabolism in vivo. PMID- 12575156 TI - [Preparation of tea polyphenols sustained-release microcapsule]. AB - With ethycellulose (EC) as matrix tea polyphenols (TP) microcapsule was prepared by using emulsion solvent evaporation technique. Effect of preparation condition on size and drug entrapment efficiency of microcapsule was investigated. We also investigated the drug entrapment effect of microcapsule. The result showed that the tawny microcapsule with highly porous surface was obtained by using polyvinyl alcohol (PVA) and sodium dodccyl sulate (SDS) as dispersant. The stirring rate, polymer concentration and drug payload affected the size and drug entrapment efficiency of microcapsule. The matrix and preparation method do not damage the ingredient of TP. The microcapsule can improve the stability of TP and sustain TP released. PMID- 12575157 TI - [GC-MS analysis of essential oil components of Alpinia oxphylla]. AB - Essential oil Components of Alpinia oxphylla were analyzed by way of applying gas chromatography, mass spetrography and computer interpretation; essential oil was abtained by supercritical CO2 fluid, thus we got 193 peaks in oil from Pot I and 210 peaks in oil from Pot II as well as identify 139 and 155 compounds respectively. PMID- 12575158 TI - [Studies on chemical constituents of the fruits of Rosa bracteata var. bracteata]. AB - The chemical constituents of Rosa bracteata Wendl. var. bracteata were studied for the first time. Six compounds were isolated from the fruits of the plant, and four of them have been identified as 5-hydroxymethyl furfural, 5-acetoxymethyl furfural, beta-sitosterol, oleanolic acid. 5-hydroxymethy furfural and 5 acetoxymethyl furfural were obtained from genus Rosa L. for the first time. Otherwise, a white wax was obtained by silica gel column chromatography, from which four main constituents were identified as nonadecene, n-docosane, hentriacontane and hexatriacontane by GC-MS. PMID- 12575159 TI - [Determination of amino acids in seeds of genus Torreya in China]. AB - OBJECTIVE: To determine the contents and analyze the ingredients of the amino acids in seeds of genus Torreya. METHODS: The amino acid contents and ingredients were determined by amino acid analyzer. RESULTS: There were 19 amino acids in the seeds of T. grandis, T. grandis cv. merrillii, T. yunnanensis, T. jackii and T. fargesii, and their amino acid contents are 9.10%, 9.90%, 5.03%, 7.26% and 6.07% respectively. There were 17 amino acids in the seeds of T. jiulongshanensis, T. nucifera and T. grandis cv. merrillii (baked) and their amino acid contents are 14.11%, 9.87% and 9.22% respectively. tively. PMID- 12575160 TI - [Determination of the saflor yellow-A in Carthamin tinctorius]. AB - The content of the saflor yellow-A in Carthamin tinctorius medicinal materials was determined by the HPLC method. The C18-ODS(150 x 4.5 mm) column and methanol acetonitrile-0.7% H3PO4(26:2:72) as a mobile phase were used. The flow rate was 1.0 ml/min and the detective wavelength was 403 nm. The average recovery was 100.72% and the relative standard deviation (RSD) was 1.73%. PMID- 12575161 TI - [Study on chemical constituents of essential oil of Biota orientalis by GC-MS]. AB - The essential oil was extracted from Biota orientalis (L) Endl by using steam distillery, analyzed with SE-54(30 m x 0.25 mm I.d) fused silica capillary column. The optimum separate and analytical conditions were researched. The amount of the components from the essential oil was determinated by monoliration method. The separated components were identified by GC-MS. There are 33 components which are make up 80% of the total essential oil were separated and identified. PMID- 12575162 TI - [A Chinese herbal compound induces neurotrophy in astrocyte by a new study method called "cerebrospinal fluid pharmacology in Chinese materia medica"]. AB - OBJECTIVE: To find a effective method by which the potency of Chinese materia medica (CMM) on the central nervous system (CNS) could be correctly investigated. METHOD: The cerebrospinal fluid(CSF) and serum after oral CMM 1 to 1.5 hours was gained and then added into the astrocyte cell culture medium. After 48 h, the above medium was in proportion transferred to the neuron culture medium. Typan blue staining, LDH and MTT test were utilized to observe the drug effect on the neuron survival. RESULTS: The serum, but not the CSF, showed the toxicity on the astrocyte. MTT test on the neuron survival showed that the OD492nm value in the CMM group was higher than the other groups (P < 0.05). The astrocyte medium stimulated by the CSF including CMM could promote the neuron axon growth. CONCLUSION: If observing the CMM potency on CNS in vitro, the CSF was better than the serum, we called it "Cerebrospinal Fluid Pharmacology in Chinese Materia Medica". Furthermore, the CMM could induce the neurotrophy of astrocyte. PMID- 12575163 TI - [Comparison of the effects of xianyu tablet and its component on electrophoretic mobility of serum LDL and expression of ET-1 mRNA and iNOS mRNA of vessel wall in atherosclerotic rabbits]. AB - Xiaoyu tablet, a compound preparation of Chinese herbal medicines, consists of Radix Salviae Miltiorrhizae(SM) and Fructus Crataegi(FC) extracts. To determine whether the proved recipe was reasonable, the effects of Xiaoyu tablet and its component on electrophoretic mobility of serum LDL and expression of ET-1 mRNA and iNOS mRNA of vessel wall in atherosclerotic rabbits were observed. The results indicated that inhibition of expression iNOS mRNA in vessel wall by Xiaoyu tablet was the same as its single extract of SM or FC, but Xiaoyu talbet was superior to SM or FC extract in reduction of electrophoretic mobility of serum LDL and inhibition of ET-1 mRNA expression in vessel wall. These results suggested that there was obvious synergism on prevention and treatment of atherosclerosis when both of the Chinese herbal medicines were simultaneously used. PMID- 12575164 TI - [Study on the extraction of isofavones in rhizoma belamcandae]. AB - In orthogonal tests, five factors were evaluted which were ethanol concentration, ethanol quantity, diameter of drug powder, reflux time and repetition times. Consideration of industrial produce, the optimal extraction was A2B2C3D2E2. That is 4-20 meshes drug powder refluxed with 70% ethanol which was 6 times weight of powder for 1 hour, repeated 2 times. PMID- 12575165 TI - [Study on genuineness of traditional Chinese medicine cortex Magnoliae officinalis]. AB - It's found that a significant correlation between the samples of Magnoliae officinalis from the provenance in phenols content and varieties or forms. The total content of magnolol and honokiol in its bark is usually used as an index to measure the quality. Therefore, Cortex Magnoliae officinalis produced in Sichuan and Hubei being considered genuinenss and good in quality, should be actually referred to local varieties. This conclusion can also reveals the nature and scientific connotation of genuineness on Cortex Magnoliae Officinalis. PMID- 12575166 TI - [Quantitative analysis of 4(3)-quinazolinedione in 4 "daqingye", leave of Isatis indigotica, Baphicacanthus cusia, Polygonum tinctorium and Clerodendron cyrtophyllum]. AB - Using 4(3)-quinazolinedione, the active constituent of "Daqingye", as the quality control criteria, 4 samples of the herbal drug collected from Isatis indigotica, Baphicacanthus cusia, Polygonum tinctorium and Clerodendron cyrtophyllum were comparetively analyzed by RP-HPLC. The results showed that the average 4(3) quinazolinedione contents (microgram/g) were 30.7, 7.2, 4.4 and 27.5 respectively. This method was proved to be quick, simple and reproducible, and provided a reliable basis for the quality control and evaluation of Chinese Traditional Drug "Daqingye". PMID- 12575167 TI - [Effect of xanthone from Canscora lucidissima on cultured myocytes anoxia reoxygenation injuries]. AB - The anoxia/reoxygenation model of cultured neonatal rats myocardial cells were developed, according to Laarse's method: anoxia and glucose deficiency for 60 min followed by reoxygenation and re-exposure to glucose for 30 min. The results showed that the survival rate of myocardial cells in the anoxia group was significantly decreased, release of lactate dehydrogenase (LDH) from myocytes was increased and membrane fluidity was decreased, all the changes were much more severe in the reoxygenation group. 30 min before anoxia, addition of xanthones: 1,8-dihydroxy-3, 5-dimethoxyxanthone, 1-hydroxy-3,5-dimethoxyanthone and 1 hydroxy-3,7,8-trithoxyxanthone, isolated from Canscora lucidissima increased the rate of myocardial cells and membrane fluidity, decreased the release of LDH. These results suggested that xanthone may provide some protective effects on the anoxia/reoxygenation damages on myocardium. PMID- 12575168 TI - [Effect of Cordyceps sinensis on the Th1/Th2 cytokines in patients with Condyloma Acuminatum]. AB - OBJECTIVE: To investigate the effect of cordyceps sinensis (CS) on the Th1/Th 2 cytokines and recurrence in patients with Condyloma Acuminatum (CA). METHOD: Double-antibody sandwich ELISA was used to study the serum level of IL-2 and IL 10 in 34 patients treated with CS(CS group) and 29 patients in control group. The recurrence rate of CA was observed. RESULT: After treated with CS, the serum levels of IL-2 and IL-10 significantly increased and decreased respectively (P < 0.05 or 0.01). No significant change occurred in control group. The recurrence times of CA in patients of CS group after electrocauterization were significantly lower than control group (P < 0.05). CONCLUSION: CS might recover the balance of Th1/Th2 cytokines and enhance Th 1 immune response in patients with CA, and reduced the recurrence of CA. PMID- 12575169 TI - [Pharmacological studies on shengenaqi oral liquid on animal model for TCM dyspnea of deficiency]. AB - The Shengenaqi oral liquid has a function of improving the inspiration and can be used in the treatment of chronic bronchitis, pulmonary emphysema and pulmonary heart disease etc. In the research, animal models for TCM dyspnea of deficiency were made by inhaling papain and injection hydrocotison into the abdominal eavity and afterwards, treated with Shengenaqi oral liquid. The effects showed: The medicine can improve the susfainability of swimming mice, increase the ability of cold tolerance and prevent the thymus from atrophy. The SOD activity in the lung tissue of mice can be improved and MDA be decreased. PMID- 12575170 TI - [Effect of ethyl acetate extract of Eclipta prostrata on mice of normal and immunosupression]. AB - OBJECTIVE: The regulatory effects of ethyl acetate extract of Eclipta prostrata (EAEEP) on immune function were studied. METHODS: The experimental immunosuppressive mice models were induced by injection of cyclophosphamide or hydrocartisone. RESULTS: The index of spleen in normal mice were decreased significantly. So was the level of hemolysin in serum, Delayed trpe hypersensitivity was also inhibited at the same condition. But EAEEP can increase all these indexes in immunosuppressive mice. CONCLUSION: EAEEP possesses obvious regulatory action on immune function in mice. PMID- 12575171 TI - [Study on extraction process for psoralen in compound prescription by orthogonal design]. AB - The study on water volume, extraction time and times for the extraction of psoralen in compound prescription Yiniao Tong capsule has been carried out by orthogonal design. The process condition has been determinated, extracting prescription herbs with water for 3 times(2 hour and 16 time amount of water all told). The determination method used in this experiment was TLC-scanning. PMID- 12575172 TI - [HPLC quantitative analysis of berberine absorpted with macroporous resin in Rhizoma Coptidis and its preparation zuo jin wan]. AB - Absorptive ability and elution program of macroporous resin to the berberine in the Rhizoma Coptidis and its preparation Zuo Jin Wan (containing Rhizoma Coptidis and Fructus Evodiae) were explored. The extract of 70% MeOH of samples were disolved in water, then alkalized to pH 12 with ammonia solution. The berberine was absorpted by the macroporous resin, the washed off water soluble impurity with alkalescent water, and eluted with 50% MeOH containing 0.5% H2SO4. The results of quantitative analysis of the berberine by HPLC were satisfied. PMID- 12575173 TI - [Herbological study for the Beimu categorical Chinese medicine on the original plants]. AB - This article reports the herbological study of "Beimu" categorical Chinese medicine on the original plants. The result showed that the medicinal "Beimu" before "Weijin" period is "Jia Beimu" (Bolbostemma paniculatum); "Nenbei" daynasty began to used "Zhe Beimu" (Fritillaria thunbergii), but "Jia Beimu" and "Hubei Beimu" (F. hupehensis) occupied an important position continuously; Last phase of "Ming" dynasty began to used the "Zhe Beimu" and "Chuan Beimu" (F. cirrhosa, F. unibracteato, F. przewalskii and F. delavayi) for "Beimu" greatly. The "Zhe Beimu" from Xiangshan, Zhejiang Province, is named "Xiang Beimu" and others are named "Tu Beimu" and the "Chuan Beimu" included the "Yi Beimu" (F. walujuewii and F. pallidiflora) continuously. So that, authors suggested that the contemporary documents such as Chinese Pharmcopoeia must recover the name "Beimu" to change the name "Tu Beimu" so as to avoid chaos with the "Tu Beimu" of the herbological documents; "Hubei Beimu" may take the place of "Zhe Beimu" but don't substitue for "Chuan Beimu"; "Yi Beimu" may be merged into "Chuan Beimu" to gear to actual circumstances and history; "Ping Beimu" to take the place of "Chuan Beimu" is not foundation from herbological documents. PMID- 12575174 TI - [The spatiotemporal change of neurotrophin family and their mRNA expression in spinal cord of cats after partial rhizotomy]. AB - OBJECTIVE: To investigate the spatiotemporal change rule of NGF, BDNF, NT-3 and their mRNA expression in spinal cord of cats after partial dorsal rhizotomy. METHODS: Rhizotomy of unilateral L1-L5, L7-S2 dorsal roots of cats was performed, leaving L6 as a spared dorsal root. By using ABC immunohistochemistry and in situ hybridization techniques, the dynamic changes of the above three factors and their mRNA in spinal lamina II of different segments were analysed. RESULTS: 1. In normal cat, NGF and it's mRNA were detected in part of neurons; BDNF, in nerve fiber terminals, varicosities and neurons; NT-3, in part of neurons, neuroglias and few nerve fiber terminals and varicosities. The mRNAs of the later two were negative. 2. The population of NGF and NGGmRNA positive neurons, NT-3 positive neurons and neuroglias increased significantly 3d-5d after rhizotomy. However, the quantity and density of positive varicosity of BDNF decreased. At 10-11d, the population of NGF and NGF mRNA positive neurons was still on the high level as that at 3-5d, and that of NT-3 began to decrease; the quantity of BDNF recovered to normal except for L, segment, but the density of positive varicosity of BDNF did not yet. The mRNAs of BDNF and NT-3 were still negative. 3. The change of each factor varied with the segments. The highest level time of NGF was earlier in L5, L6 than in L3; the recovery of the quantity of BDNF was the latest in L7; the change of NT-3 positive neuroglia was the same at each segment, but the number of NT-3 positive neuron in L5, L7 returned to normal at 10-11d, and that of L3 did not. CONCLUSION: The three factors all play roles in spinal plasticity after partial rhizotomy, but they function at different time phase and last different time length. PMID- 12575175 TI - [Construction of DNA vaccine including a chimeric gene encoding flagellin and outer membrane protein antigen from Leptospira interrogons serovar lai]. AB - OBJECTIVE: To construct a fusion expression vector for DNA vaccine, including a flagellin gene (flaB2) and an outer membrane protein gene (ompL1) to enhance the protective immunity of Leptospira interrogans serova Lai (Stain 017). METHODS: The DNA fragments encoding flaB2 and ompL1 were amplified respectively by PCR and then fused. The obtained chimeric gene ompL1-flaB2 contained a synthetic linker of ten amino acids to sustain its space conformation. RESULTS: Identified by restriction enzyme analysis, an insert fragment of 1.8 kb was demonstrated. FlaB2 and ompL1 respectively had the same sequence as reported by DNA sequencing analysis. CONCLUSION: A fusion expression plasmid containing flaB2 and ompL1 was constructed. PMID- 12575176 TI - [Molecular cloning and expression of the immunodominant protein Ag85A from Mycobacterium tuberculosis H37Rv strain]. AB - OBJECTIVE: To provide the target gene and target antigen for the development of new vaccine against tuberculosis. METHODS: According to the gene sequence encoding protein Ag85A from Mycobacterium tuberculosis H37Rv strain, we designed a pair of oligonucleotide primers, obtained the gene by using polymerase chain reaction, and inserted the gene into the BamH I and EcoR I site of plasmid pBK CMV to construct recombinant plasmid, and after that, the recombinant plasmid was transferred into E. coli XL1-Blue MRF' and induced with IPTG. The expression product of the gene was analyzed by using SDS-PAGE and western-blotting. RESULTS: The gene encoding the protein Ag85A of Mycobacterium tuberculosis H37Rv strain was successfully amplified by using PCR. A recombinant shuttle plasmid was constructed. The recombinant plasmid stably expressed recombinant Ag85A protein relative molecular mass 32 x 10(3) in E. coli XL1-Blue MRF'. CONCLUSION: A recombinant plasmid which contains the gene encoding the protein Ag85A of Mycobacterium tuberculosis H37Rv strain has been successfully constructed. The recombinant plasmid can stably express recombinant protein relative molecular mass 32 x 10(3) in E. coli XL1-Blue MRF'. These results could serve as a basis for further studies on the usefulness of the gene and its expression product in the development of new vaccine against tuberculosis. PMID- 12575177 TI - [Comparison of the immune response to gene transfer into the rat brain with helper or helper virus-free HSV-1 vectors]. AB - OBJECTIVE: This study was conducted to compare the immune response to gene transfer with helper virus or helper virus-free vectors. METHODS: Two vector stocks were delivered by stereotactic injection into the striatum and then the immune response markers including TCR, CD4, CD8, MHC-I and MHC-II were examined. RESULTS: The data showed that the immune response to either vector stock reached maximal levels by 4 to 7 days after gene transfer. However, the maximal levels of all markers to the helper virus-free stock were 3 to 10 folds lower than the markers to the helper virus stock. The immune response to the helper virus-free stock declined rapidly to be close to the levels of PBS control. CONCLUSION: The immune response to the helper virus-free stock is significantly less than the response to the vector stock that contains helper virus. PMID- 12575178 TI - [Expression of hepatitis C virus envelope proteins with a recombinant baculovirus expression system]. AB - OBJECTIVE: To acquire stable expression of envelope proteins of hepatitis C virus in insect host cells and use the expressed envelope proteins for detecting the serums of patients with hepatitis C. METHODS: The envelope gene of HCV H strain was amplified by PCR and inserted in baculovirus vector BacPAK8, and then recombined with linear BacPAK6 DNA in insect cells. The recombinant baculoviruses were selected by the plaque assay. The insect cells were infected by the recombinant baculoviruses that contained the target gene produced E1, E2 proteins, which were characterized with the immunoblot assay and the immunofluorescence and were used to determine 35 serum samples of patients with hepatitis C. RESULTS: The expressed E1, E2 proteins showed that the relative molecular mass of E1 is about 21 x 10(3) and 33 x 10(3), and that of E2 is about 60 x 10(3). Detection of immunofluorescence indicated that E1, E2 proteins are localized in the cytoplasm of the infected cells. Four of the 35 serums responded to expressed E1; one of them was found to recognize E2 protein. Three of 9 serums which were HCV RNA positive by PCR testing got united to E1, E2. CONCLUSION: The HCV envelope protein can be expressed stably in the insect cells. Expressed E proteins could be used in the serologic analysis of the patients' serums. PMID- 12575179 TI - [The role of progesterone in the regulation of gene expression of insulin-like growth factor-I receptor in human decidual stromal cells of early pregnancy in vitro]. AB - OBJECTIVE: To assess the effect of progesterone on the gene expression of insulin like growth factor-I receptor (IGF-I R) in human decidual stromal cells of early pregnancy in vitro. METHODS: Semiquantitative reverse transcriptase polymerase chain reaction, using beta-ACTIN as internal standard, was applied to determine the levels of IGF-I R mRNA in human decidual stromal cells of early pregnancy in vitro after cultured with different concentrations of progesterone for 72 hours or cultured with 0.1 mumol/L of progesterone for different periods of time. RESULTS: The expression of IGF-I R mRNA was significantly positive in human decidual stromal cells of early pregnancy in vitro. The levels of IGF-I R mRNA were down-regulated by progesterone, and showed significant negative-correlation with the concentration of progesterone (r = -0.680, P < 0.001). The levels of IGF I R mRNA showed no significant correlation with time when the final concentration of progesterone was 0.1 mumol/L (r = 0.005, P > 0.05). CONCLUSION: Progesterone may play an important role in the regulation of proliferation and decidualization of stromal cells by down-regulating the expression of IGF-I R mRNA in human decidual stromal cells of early pregnancy, which is important for the maintenance of early pregnancy. PMID- 12575180 TI - [Gene expression of insulin-like growth factors in human villous trophoblast cells of early pregnancy in vitro]. AB - OBJECTIVE: To investigate the gene expression of insulin-like growth factors(IGFs) in human villous trophoblast cells of early pregnancy in vitro and their action. METHODS: Semiquantitative reverse transcriptase polymerase chain reaction(RT-PCR), with beta-ACTIN as internal standard, was applied to determine the expression of IGFs messenger RNA in human villous trophoblast cells of early pregnancy in vitro. RESULTS: The expression of IGF-II mRNA, IGF-I R mRNA and IGFBP-3 mRNA was detectable in human villous trophoblast cells of early pregnancy in vitro. CONCLUSION: IGFs appear to play important regulation roles in early invasion, proliferation and differentiation of cytotrophoblast, and in the formation of placenta and the development of embryo via autocrine and/or paracrine way. PMID- 12575181 TI - [Discharge patterns of non-motoneurons in the facial nucleus of rats]. AB - OBJECTIVE: To explore the discharge patterns of non-motoneurons in the facial nucleus of rats. METHODS: After retrograde degeneration of the facial motoneurons induced by section of the facial nerve, the discharge of non-motoneurons in the facial nucleus was extracellularly monitored, and their phase relations to the respiratory cycle were investigated. RESULTS: In 18 rats, totally 142 non motoneurons with spontaneous discharge were recorded in the facial nucleus. Among them, 127 (89.4%) were non-respiratory related neurons (NRRNs) and 15 (10.6%), respiratory related neurons (RRNs). CONCLUSION: There exist different kinds of nonmotoneurons in the facial nucleus. The results of this study suggest that the facial nucleus not only controls the movement of the facial skeletal muscles, but also participates in the regulation of other functions such as respiration. PMID- 12575182 TI - [Antisense inhibition of gene expression in human dendritic cells by peptide nucleic acid against CD86]. AB - OBJECTIVE: Dendritic cells (DCs) are the most potent antigen presenting cells (APCs) of the immune system. We intent to block the expression of CD86 in DCs using antisense peptide nucleic acids (PNA), a novel synthetic structural DNA mimic, and interrupt the second signal transmission so that a suppression of corresponding T cell function can be achieved. METHODS: Human DCs grown up from peripheral blood monocytes in GM-CSF and IL-4 were collected. We investigated antisense PNA internalization with laser scan confocal microscope (LSCM). Fluorescence immunocytochemistry, flow cytometry and RT-PCR were used to determine the expression of CD86 protein and mRNA in DCs. RESULTS: LSCM proved that cultured immature DCs could internalized PNA efficiently, according to the specific internalization property of the immature DCs. Antisense PNA DC exhibited striking reductions in cell surface staining for CD86, but not MHC class II, and were poor stimulators of T cell proliferation. RT-PCR found that PNA depressed the amounts of CD86 mRNA in DCs. CONCLUSION: Antisense PNA against CD86 could inhibit the expression of CD86 mRNA and protein in DCs. The blockade of B7/CD28 pathway may increase the potential of costimulatory molecule-deficient antisense PNA DCs of donor origin to induce long-lasting allograft survival. PMID- 12575183 TI - [The difference in proliferation and differentiation ability of cultured marrow stromal cells between 3-month-old and 12-month-old rats in vitro]. AB - OBJECTIVE: To compare the difference in proliferation and differentiation (to osteoblast or adipocyte) ability of cultured rat marrow stromal cells (rMSCs) between 3-month-old and 12-month-old rats in vitro. METHODS: The rMSCs of 3-month old and 12-month-old were cultured in vitro. The growth curves were depicted to compare their proliferation ability. Both 3-month-old and 12-month-old rMSCs were induced to osteoblasts or adipocytes by osteogenic inducer or adipogenic inducer. At different times, these cells were observed by histochemistry staining. RESULTS: The growing ability of both passage 4 (P4) and passage 9 (P9) of 12 month-old rMSCs decreased in comparison with that of 3-month-old rMSCs. The growing ability of P9 was less than that of P4 of 12-month-old rMSCs. The expression of alkaline phosphatase (ALP) of the control group and induced group of 12-month-old rMSCs was less than that of 3-month-old rMSCs after 1-week osteogenic induction. 12 days later, calcification was observed in 3-months-old group. Lipid droplets occurred in the cells of 12-month-old group after 2-day adipogenic induction, whereas the droplets occurred after 3-day or 4-day induction in the 3-month-old group. CONCLUSION: The ability of proliferation and osteogenesis of 12-month-old group is weaker than that of 3-month-old group, but the ability of adipogenesis of 12-month-old group is stronger. PMID- 12575184 TI - [Osteocalcin and type I collagen mRNA expression of rat marrow stromal cells under induction condition]. AB - OBJECTIVE: To investigate osteocalcin and type I collagen mRNA expression of rat marrow stromal cells under induction condition. METHODS: Cultured rat marrow stromal cells were induced by osteogenic compound(composed of dexamethasone 10( 3) mol/L, beta glyceralphosphate 10 mmol/L, ascorbic acid 50 micrograms/ml). Osteocalcin and type I collagen mRNA expression were measured using RT-PCR technique. RESULTS: It was found that rat marrow stromal cells in induced group have an increased time-dependent expression of osteocalcin and type I collagen mRNA, in comparison with control groups. And the necessity of 1,25-(OH)2D3 in induced osteocalcin mRNA expression was confirmed. CONCLUSION: The cultured rat marrow stromal cells kept the undifferentiated state and had osteogenic potential testified by increased osteocalcin and type I collagen mRNA expression when induced. PMID- 12575185 TI - [The expression of collagen type I and type II during the second fracture healing]. AB - OBJECTIVE: This study was conducted to disclose the rules of collagen type I and II production and hence to acquire knowledge about the mechanism of repairing during the second period of fracture healing. METHODS: An experimental model of second fracture healing at rat's femurs was used to study the production of collagen type I and II by means of immunohistochemistry; meanwhile the histologic changes were observed with the use of HE staining and Alcian blue staining at 3 days, 1, 2, 3, 4 weeks postoperatively. RESULTS: The expression of both collagen type I and II was detected by immunohistochemistry at 1 week. The peak expression of collagen type II occurred at 3 weeks. The higher expression of collagen type I was observed at 4 weeks. CONCLUSION: The morphologic changes of chondrocyte and the production of collagen type I and II by chondrocyte in cartilage callus are correlated with their repairing function during different periods of fracture healing. PMID- 12575186 TI - [An in vivo study of reseeding endothelial cell for attenuation of hyperacute rejection after xenotransplantation]. AB - To investigate the possible significance and usefulness of reseeding the endothelial cell of recipient for the prevention of hyperacute rejection (HAR) resulting from the xenograft. METHODS: Discordant xenotransplantation model (guinea pig-to-rat) was adopted in this study. Firstly endothelial cells from rat abdominal aorta were separated and cultured. Secondly the guinea pig abdominal aorta of which the endothelium had been removed was cultured with the suspension containing rat endothelial cells (4 x 10(6)/ml). The viability of cultured vessel was assessed using light microscopy and transmission electron microscopy. Thirdly, the guinea pig vessels reseeded with rat endothelial cells were transplanted to rat in vivo. The thrombrosis time was observed. The condition of deposit of IgM and C3 in the recipient's transplanted vessel endothelium was examined by immuno-fluorescence staining assay. RESULTS: The thrombrosis time of the guinea pig vessel reseeded with rat endothelial cells (20.3 + 4.42 h) was significantly prolonged as compared with that of the normal guinea pig vessel (0.35 + 0.284 h) and the guinea pig vessel that had been deprived of endothelium (0.165 + 0.77 h). No deposit of IgM and C3 was seen in the new endothelium of guinea pig vessel in the treated group, whereas deposit of IgM and C3 was observed in the untreated normal guinea pig aorta. CONCLUSION: Donor vessels reseeded with endothelial cells from the recipient will undergo less severe rejection and this technique may be very useful in the attenuation of HAR. PMID- 12575187 TI - [Effect of para-aminobenzoic acid on the growth of Actinomyces viscosus]. AB - OBJECTIVE: To examine the effects of para-aminobenzoic acid (PABA) on the growth of Actinomyces viscosus. METHODS: Different concentrations of PABA (10(-10)-10( 3) g/L) were each transferred to modified Carlsson medium. Actinomyces viscosus ATCC19246 grew in them. And the cultures were incubated at 37 degrees C anaerobically in the atmosphere of 80%N2, 10%H2, 10%CO2 for 48 h. Actinomyces viscosus OD values (lambda = 540 nm) were obtained with UV-1601. Colony forming unit (CFU) was established by growth of Actinomyces viscosus in culture when different concentrations of PABA (10(-10)-10(-3) g/L) were present. RESULTS: Different concentrations of PABA (10(-10)-10(-4) g/L) had different stimulating effects on the growth of Actinomyces viscosus (P < 0.05). But this kind of stimulating effect declined when PABA concentration was 10(-5) g/L, and as PABA (10(-3) g/L) was present, this kind of effect was missing. CONCLUSION: The phenomena indicated that PABA has stimulating effect on the growth of Actinomyces viscosus, particularly when PABA is at the concentration of 10(-6) g/L. PMID- 12575188 TI - [Relationship between the infiltration arts and penetration depth of molten glass into aluminum oxide matrix]. AB - OBJECTIVE: To explore the influences of infiltration time and temperature on the penetration depth of GI-II glass into alumina matrices, thus providing a theoretical basis for facilitating the clinical fabrication of all-ceramic restorations. METHODS: After preparing cuboid alumina specimens 7 x 7 x 5 mm3 in size, we carried out the infiltration firing procedure at 1100 degrees C for six different time duration (30, 60, 90, 120, 150, 180 minutes), and at five different temperatures (1060, 1080, 1100, 1120, 1140 degrees C) for 1 hour each. The penetration depth of glass into the alumina matrices was measured under a stereoscope with magnification of 10 times. RESULTS: The infiltration time duration and penetration depth were not linearly but exponentially related for both Vita and GI-II materials (r = 0.9886 for Vita and r = 0.9932 for GI-II). The regression equations were as follows: d2 = 0.1122t - 0.4955 (Vita) and d2 = 0.1638t + 0.5873 (GI-II). The temperature and depth were not linearly related either, with increased penetration speed under higher temperatures for GI-II material; the tendency for Vita material was just the opposite. CONCLUSION: At an infiltration temperature of 1100 degrees C, the molten infiltration time duration of GI-II glass could be reduced to 1 hour for copings and 3 hours for anterior bridge substructures due to its better infiltration ability. PMID- 12575189 TI - [A study on the protective mechanism of yisheng injection against the anoxia reoxygenation injury to endothelial cells]. AB - OBJECTIVE: To explore the mechanism of ischemia reperfusion injury (IRI) and the protective effect of Yisheng injection on endothelial cells (ECs). METHODS: Human umbilical veins endothelial cell line ECV304 was cultured in RPMI medium 1640 without glucose under 100% N2 for one and a half hours, and then in RPMI medium 1640 with glucose under normal conditions for 5 hours to mimic ECs' IRI posttransplantaion. A pure nature medicine, Yisheng injection was added into the medium in experimental group. Intracellular calcium and mitochondrial were shown through flourescent staining. The activities of succinic dehydrogenase (SDH) and lactic dehydrogenase (LDH) were detected by cytochemical staining. The concentrations of superoxide dismutase (SOD), malonaldehyde (MDA) and nitrous oxide (NO) in culture medium were determined. RESULTS: After treatment with anoxia-reoxygenation, the ECs' morphologic changes were observed, and intracellular calcium concentration, LDH activity, MDA concentration became higher. Meanwhile, mitochondrial membrane potential, concentrations of SOD and NO were lower. Yisheng injection made these changes disappear or become less. CONCLUSION: Anoxia-reoxygenation induces lipid peroxidation, calcium superrcharge in ECs, and damage to mitochondrial function. Yisheng injection can reverse these changes, thus protecting the endothelial cells. PMID- 12575190 TI - [Cultivation of fibroblasts derived from urethral scar and the effects of salviae miltiorrhizae and ligustrazine on its growth]. AB - OBJECTIVE: To explore the possible effect and related mechanism of salviae miltiorrhizae (SM) and ligustrazine in the treatment of urethral scar. METHODS: In vitro cultivation of fibroblasts derived from urethral scar has been developed. The effects of these two medicines on its morphology and proliferation have been detected based on the in vitro culture system. RESULTS: The spindle cells were found to become elliptical in shape. The two medicines inhibited the proliferation of fibroblasts derived from urethral scar, and this may be resulted from their inhibitive effects on cell division rather than cell necrosis or degradation. CONCLUSION: Salviae multiorrhizae and ligustrazine as effective scar inhibitors may have application prospects in the treatment of urethral stricture. PMID- 12575191 TI - [Culture of mesangial cells in two different states and a study on their functions]. AB - OBJECTIVE: To make a comparision of cell function between the mesangial cells from rats with diabetic mellitus (DMC) and the mesangial cells stimulated by high glucose (HMC) and to explore the feasibility and advantage of culturing mesangial cells (MC) from diabetic rats in studies on the pathogenesis of diabetic nephropathy. METHODS: DMC were obtained from rats with streptozotocin-induced diabetes and were cultivated in normal medium. The MC from normal rats were divided into HMC high glucose and NMC cultured in normal medium as control. The cellular proliferation was assessed using 3H-thymidine incorporation. Production of fibronectin (FN) was analysed by ELISA. And with the use of laser scanning confocal microscopy, the calcium concentration ([Ca2+]i) and the response of [Ca2+]i to Angiotensin II (Ang II) were measured. RESULTS: The 3H-TdR incorporation efficiency of DMC was significantly higher than that of CMC; the 3H TdR incorporation efficiency of HMC was lower than that of CMC (P < 0.05). The secretion of FN in DMC and HMC was higher than that in CMC (P < 0.01 and P < 0.05 respectively). The response of [Ca2+]i to Ang II in DMC and HMC was decreased (P < 0.05), and the decrease was more significant in DMC than in HMC. CONCLUSIONS: The function and biologic character of mesangial cells cultured from diabetic rats are more similar to those of mesangial cells from diabetic in vivo. The result suggest that establishing a diabetes model first and starting the culture of MC next is probably a good method for investigating the mesangial cells in diabetic nephropathy. PMID- 12575192 TI - [Effects of testosterone on VEGF and FLK-1 protein expression in the ventral prostate of rat]. AB - OBJECTIVE: To investigate the effects of testosterone on VEGF and FLK-1 protein expression in the ventral prostate of Rat. METHOD: SD rats were castrated, and 7 days later they were injected with different doses of testosterone. The histological changes were observed with the use of HE staining, and the protein expression of VEGF, FLK-1 were measured by immunohistochemical test. RESULTS: In the normal ventral prostate, different growth forms of epithelium were observed. In the castrated group, the proteins of VEGF, FLK-1 were positive in growth active epithelium and vascular endothelium; the weight of prostate decreased (P < 0.05); the histological appearance was epithelial atrophy, and there was significantly decreased protein expression of VEGF, FLK-1 in the prostatic epithelium (P < 0.05). It was found that testosterone injection stimulated prostatic hyperplasia gradually The histological changes were epithelial progressive proliferation. And a gradually increased protein expression of VEGF, FLK-1 in the prostatic epithelium was detected (P < 0.05). CONCLUSION: Testosterone can stimulate prostatic growth probably by upregulating the protein expression of VEGF and FLK-1. PMID- 12575193 TI - [The paraoxonase Gln-Arg 192 polymorphism in patients with endogenous hypertriglyceridemia in Chinese population]. AB - OBJECTIVE: To investigate the 192 Gln-Arg polymorphism of paraoxonase (PON) gene and its relationship with serum lipids and apolipoproteins (apo) levels in patients with endogenous hypertriglyceridemia in Chinese population in Chengdu area. METHODS: The genotype and allele frequencies of paraoxonase gene 192 Gln Arg polymorphism were assayed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). Serum lipids were measured by enzymatic kits and apolipoproteins AI, A II, B100, C I, C II and E were measured by the RID kits developed by the Apolipoprotein Research Unit of this university in 128 HTG patients whose fasting serum TG levels were > or = 2.26 mmol/L and 129 healthy subjects whose fasting serum TG levels were < 1.82 mmol/L and TC levels < 6.2 mmol/L from a population of Chinese Han nationality in Chengdu area. RESULTS: Both in HTG group and control group, the QR genotype of PON gene was the major one, and the frequencies were 0.515 and 0.581 respectively. No differences were found in PON gene Gln-Arg polymorphism between the HTG group and the control group. In the control group, the QQ genotype of PON gene was found to have higher serum apoA I levels, compared with the RR genotype (P < 0.05). But in the HTG group, when compared with the RR genotype, the QQ genotype was found to have lower serum apoA I and A II levels and higher serum apoE levels. CONCLUSION: These may be an association of the QQ genotype of the paraoxonase 192 Gln-Arg polymorphism with the decrease of serum apoA I level and the increase of serum apoE level in endogenous hypertriglyceridemica. PMID- 12575194 TI - [Angiotensinogen and angiotensin-I converting enzyme gene variations in Chinese pregnancy induced hypertension]. AB - OBJECTIVE: This study was designed to observe the genotype and allele frequency of angiotensinogen (AGT) and angiotensin-I converting enzyme (ACE) genes in pregnancy induced hypertension (PIH) and compare them with those in normal pregnant women. METHODS: Eighty-one women with pre-eclampsia (median age 28 years) and 205 normal pregnant women as controls (median age 28 years) from a population of Chinese Han nationality in Chengdu area participated in this study. The pregnant patients whose blood pressure exceeded 140/90 mmHg (or 18.7/12 kPa) were recruited with a rigorous definition of pre-eclampsia. Genotyping was performed using PCR-amplified DNA fragment for ACE I/D and PCR-RFLP for AGT polymorphism. RESULTS: The allele frequencies of AGT235T and ACE D in PIH and normal pregnant women were 0.856 vs. 0.846 and 0.333 vs 0.364, respectively. No significant difference in these allele frequencies was observed between PIH and normal pregnant women. The allele frequencies of ACE D in Caucasians were higher than those in Chinese population (P < 0.001). The allele frequencies of AGT235T in Caucasians (0.350-0.482) and other populations (0.620-0.739) were lower than that in Chinese (0.846) (P < 0.01, P < 0.001). CONCLUSION: There was no evidence in this study for the association of AGT M235T and ACE I/D polymorphism with pre eclampsia in Chinese population in Chengdu area. PMID- 12575195 TI - [The alteration of inflammatory cytokine during acute pancreatitis]. AB - OBJECTIVE: To observe the alteration of both inflammatory and anti-inflammatory cytokines during acute pancreatitis, and to investigate the effect of somatostatin on modulation of inflammatory and anti-inflammatory cytokines in experimental acute pancreatitis. METHODS: SD male rats were divided into 3 groups: group 1. the normal rats as control (n = 6); group 2, the rats with acute pancreatitis induced by transabdominal injection of 5% sodium cholate sulfur (at the volume of 1.0 ml/kg) into the parcreatic duct and not given drug treatment; group 3, the rats injected with stilamin 20 micrograms/kg, intravenously, 30 minutes after the successful induction of acute pancreatitis. The animals in groups 2 and 3 were killed at 2, 6 and 24 hours after operation. The blood samples were taken for measurement of IL-1, TNF alpha, IL-6 (by Bioassay) and IL 10, TNF-beta (by ELISA). The wet weight of pancreatic tissue and amylase were also determined. RESULTS: Serum IL-1, TNF alpha, IL-6, IL-10 and TGF-beta in control group were 0.56 +/- 0.06 ng/ml, 23.50 +/- 1.87 IU/ml, 69.0 +/- 6.40 IU/ml, 32.05 +/- 14.87 pg/ml and 66.4 +/- 13.20 pg/ml, respectively. After acute pancreatitis was induced, the serum level of these inflammation-concerned cytokines increased significantly in group 2 (P < 0.05). Serum IL-1, TNF alpha, IL-6, IL-10 and TGF-beta in group 2 at 24 hours after pancreatitis were 1.15 +/- 0.13 ng/ml, 55.33 +/- 12.79 IU/ml. 127.17 +/- 13.91 IU/ml, 68.13 +/- 19.90 pg/ml, and 103.77 +/- 28.95 pg/ml, respectively. After administration of somatostatin, the inflammation-concerned cytokines in group 3 were remarkably decreased (P < 0.05). Serum IL-1, TNF alpha, IL-6, IL-10 and TGF-beta in group 3 were 0.83 +/- 0.12 ng/ml, 33.00 +/- 7.40 IU/ml. 71.83 +/- 6.34 IU/ml, 42.2 +/- 14.55 pg/ml, and 45.98 +/- 18.10 pg/ml, respectively. The indeies of the severity of pancreatitis, such as amylase and the weight of pancreas alse improved in group 3. CONCLUSION: Both inflammatory and antiinflammatory cytokines increased remarkably in the rats with acute pancreatitis. This result indicates that there is a potential tendency of inflammatory response syndrome and compensatory anti-inflammatory response syndrome in the course of acute pancreatitis. Somatostatin can modulate the derangement of these cytokines in acute pancreatitis. PMID- 12575196 TI - [The expression of glucocorticold receptor isoform in peripheral blood mononuclear cell with Graves' ophthalmopathy]. AB - OBJECTIVE: To study the expression of glucocorticoid receptor (GR) isoform in the peripheral blood mononuclear cell (PBMC) of patients with Graves' ophthalmopathy (GO). METHODS: Semi-quantitative RT-PCR was used to analyze RNA isolated from PBMC of the patients with GO and the normal volunteers. The level of plasma total cortisol (PTC) at 8 a.m. was measured. RESULTS: The hGR alpha/GR beta mRNA ratios of the two groups were 7.58 +/- 5.42 and 14.65 +/- 8.30, respectively. There was significant difference between the two groups (P < 0.05). The PTC levels of the two groups were 304.23 +/- 92.06 and 313.73 +/- 111.05, and no significant difference between them was noted (P > 0.05). No correlation was found between hGR alpha/GR beta mRNA and PTC levels. CONCLUSION: hGR alpha/GR beta mRNA may play a role in the pathogeny of Graves' ophthalmopathy. PMID- 12575197 TI - [Study on influencing factors of the by-product CHCl3 produced in the electrochemical sterilization process]. AB - OBJECTIVE: To gain an understanding of the progressive course and influencing factors of the by-product CHCl3 produced in the electrochemical sterilization process. METHODS: Taking filtering water, using graphite and Ti(matrix)-Ti as electrodes, adding different concentrations of SO4(2-) and Cl- in the filtering water, adjusting the electric current for analyzing sample in different times, and reviewing the production of CHCl3 under different conditions. RESULTS: The quantity of CHCl3 produced in the electrolysis using Ti (matrix)-Ti was greater than that using graphite. Besides, it was found that the more the current density increased, the more would be the production of CHCl3. No significant difference in the quantity of CHCl3 was seen after the addition of trace electrolyte Na2SO4; however, after the addition of trace electrolyte NaCl, (CHCl3) increased with the increase of (Cl-). CONCLUSION: The graphite electrode should be the best and choicest electrode for use in the electrochemical sterilization of drinking water; the time of electrolysis should not be more than 10 minutes; the appropriate current density current is 1 mA/cm2. These suggestions may conduce to minimizing the production of CHCl3. PMID- 12575198 TI - [The preparation procedure of TiAdapt abutment effect on the implant to abutment joint stability]. AB - OBJECTIVE: This study was designed to determine the difference in the implant to abutment joint stability resulting from the preparation procedure in adjusting the TiAdapt abutment. METHODS: Thirty Branemark implants (10RP, 10NP, 10WP) and 10 TiAdapt RP, TiAdapt NP and TiAdapt WP abutments formed the experimental populations of 3 groups. Samples in each groups were further divided into subgroups A and B. Subgroup A was operated with preparation procedure whereas the other served as control. The torque required and the gap between abutment and implant before and after preparation were measured. RESULTS: There were no significant differences in group RP and group WP; there were significant differences in group NP at the significance level of 0.05. CONCLUSION: These results indicate that the implant to abutment joint stability has been strengthened by the abutment preparation procedure. PMID- 12575199 TI - [Observation on the inhibiting effect of fluor protector on the demineralization of bovine enamel after exposure to some beverages]. AB - OBJECTIVE: To investigate the role of Fluor Protector in the demineralization of bovine enamel after exposure to some beverages. METHODS: Twenty-four bovine teeth were divided into experimental and control groups. The enamel specimens of the experimental group were pretreated with Fluor Protector and then exposed to beverages; the enamel specimens of the control group were exposed to beverages directly. All the enamel specimens were exposed to beverages 10 times a day and 5 minutes each time. After 7 days' exposure to the beverages, all the specimens were observed by scanning electron microscope (SEM). RESULTS: Different degrees of solubilization of enamel prisms in the control group were demonstrated by SEM. The solubilization of enamel prisms in the experimental group was much more slight than that in the control group. CONCLUSION: Fluor Protector can inhibit the demineralization of enamel caused by beverages. PMID- 12575200 TI - [The in vitro study of the effects of 11 kinds of traditional Chinese medicine on the growth and acid production of Actinomyces viscosus]. AB - OBJECTIVE: To assess the effects of different natural medicines on the growth and acid production of Actinomyces viscosus, thus making preparations for screening an effective agent to mediate the balance of oral microflora. METHODS: Actinomyces viscosus ATCC 19246 was chosen as the experimental bacteria. 11 kinds of traditional Chinese medicine, such as Rhizoma Ligustici Chuanxiong, Sargentodoxa Cuneata and Galla Chinensis were extracted by means of maceration, percolation and reflux extraction. First, the values of MIC of various extracts were measured. Second, the experimental medium containing various extracts was prepared. The concentration of the extracts was lower than the MIC of the medicine, and the initial pH of the medium was 7.4. Then Actinomyces viscosus was cultured in the medium for 48 h, and finally the rest pH was measured. RESULTS: When the concentration of the medicines was lower than or equal to 8.000 mg/ml, it was found that all kinds of medicine except Radix Notoginseng can inhibit the growth of Actinomyces viscosus effectively, especially Polistes mandarinus and Semen Arecae. Tea polyphenols, Radix Notoginseng, Radix et Rhizoma Rhei, Polistes mandarinus and Sargentodoxa cuneata can inhibit the acid production of Actinomyces viscosus effectively, but Radix Scutellariae, Rhizoma Ligustici Chuanxiong, Semen Arecae, Radix Angelicae Dahuricae, Galla Chinensis and Catechu have no preliminary effect on it. CONCLUSION: Tea polyphenols, Radix et Rhizoma Rhei, Polistes mandarinus and Sargentodoxa cuneata can inhibit the growth and the acid production of Actinomyces viscosus effectively. PMID- 12575201 TI - [An epidemiological study on senile cataract in urban and rural areas of Chengdu]. AB - OBJECTIVE: To find out the status and distribution characteristics of senile cataract, which is one of the common disease affecting the quality of life of the elderly people. METHODS: One thousand and sixteen people aged 60 or more in the urban and rural areas of Chengdu were included by cluster random sampling and investigated by questionnaire method and medical examination. RESULTS: It was found that the prevalence of senile cataract was 52.3% in urban area and 51.3% in rural area. The prevalence was higher in women than in men, and it increased with age. The prevalence of senile cataract by past diagnosis was lower than that by present medical examination; this difference was found obvious in the rural area. CONCLUSION: The prevention and treatment of senile cataract should be strengthened. PMID- 12575202 TI - [A study on the influence factors for social adaptive behavior of children]. AB - OBJECTIVE: To investigate the influence factors for social adaptive behavior of children. METHODS: Six hundred children of 6 to 12 years old were investigated with the adaptive behavior assessment form of American children (AAMD-ABS revised edition). The influence factors for social adaptability of children were analyzed by stepwise regression. RESULTS: It was found that preschool fostering type, parents' relation, social mood of living area, and mother's age and health state enforced a remarkable influence on the social adaptive behavior of children. CONCLUSION: A good fostering type, parents' harmonious relationship, health children's social environment and good health of mother would help improve children's social adaptability. PMID- 12575203 TI - [The effects of activities of Ca(2+)-ATPase and Na(+)-K(+)-ATPase in plasma membranes of hepatocytes on the formation of calcium bilirubinate gallstone]. AB - OBJECTIVE: To investigate the effects of activities of Ca(2+)-ATPase and Na(+) K(+)-ATPase in plasma membranes of hepatocytes on the formation of calcium bilirubinate gallstone. METHODS: The rabbit models for studying calcium bilirubinate gallstone were used. One hundred and three rabbits were randomly divided into the control (sham operation) group (Con, n = 28), the simple biliary obstruction group (BO, n = 36), and the biliary obstruction and infection group (BOI, n = 39). The activities of Ca(2+)-ATPase and Na(+)-K(+)-ATPase in plasma membranes of hepatocytes and the intracellular calcium content were measured on the 3rd, 7th, 14th and 20th days after operation. RESULTS: The activities of Ca(2+)-ATPase and Na(+)-K(+)-ATPase decreased remarkably in all phases of BOI and BO groups as compared with those of Con group (P < 0.01). The over-loaded intracellular calcium was found in both BOI and BO groups. The above-mentioned changes were more significant in BOI group than in BO group (P < 0.05). CONCLUSION: The progressive decrease of the activities of Ca(2+)-ATPase and Na(+) K(+)-ATPase is in close relationship with the continuous increase of intracellular calcium content during the formation of calcium bilirubinate gallstone in rabbit models. Infection can aggravate those changes and further the formation of stone. PMID- 12575204 TI - [The apoptosis of human gastric cancer cell induced by Acanthopanax giraldii Harms var Hispidus Hoo Polysaccharides in vitro]. AB - OBJECTIVE: To investigate the effect of Acanthopanax Giraldii Harms Var Hispidus Hoo polysaccharides (AGP-I) on the human gastric cancer cell. METHODS: DNA agarose gel electrophoresis and flow cytometry techniques were adopted. RESULTS: After treatment with AGP-I, apoptotic peaks in cell cycle analysis and DNA ladder on the agarose gel were observed. CONCLUSION: The results indicate that AGP-1 may inhibit cancer by inducing cancer cell apoptosis. PMID- 12575205 TI - [Comparison between apoptosis cells count and ATP-bioluminescence assay in ovarian cancer chemosensitivity testing in vitro]. AB - OBJECTIVE: This study was aimed to evaluate the potentiality for the use of apoptosis cells count as a chemosensitivity testing in vitro for ovarian cancer. METHODS: An end-labelling assay was used to determine the apoptosis cells of 4 specimens of fresh ovarian cancer cells and the results were compared with that of ATP assay. RESULTS: It was found that chemotherapy can induce apoptosis, and the same chemotherapeutic agent can induce different apoptosis cells for different cancers. CONCLUSION: The observed agreement between apoptosis cells count and ATP assay was 0.75. The above data demonstrate that apoptosis cells count can be used as a chemosensitivity testing for ovarian cancer, and a combination of these two methods may improve the clinical effects of chemotherapeutic agents. PMID- 12575206 TI - [Experimental reconstruction of extensive anterior defect of rabbit trachea with the use of free auricular cartilage]. AB - OBJECTIVES: To determine whether free auricular cartilage grafts can be used to reconstruct the extensive anterior defect of rabbit trachea and observe the difference between autograft and allograft. METHODS: Twenty New Zealand white rabbits were divided into autograft group (n = 10) and allograft group (n = 10). All grafts were taken from the right auricle, and defect included 8 to 10 rings of trachea. The gross morphorlogical features, endoscopic examinations, biomechanic determinations and histological findings of grafts were assessed at 1,2,4,8 and 12 weeks after operation. RESULTS: Eighteen rabbits survived. Mild tracheal stenosis was observed under endoscope. The maximum stress per mm at 0,4,8 and 12 weeks was 2.54 +/- 0.19, 1.31 +/- 0.21, 1.72 +/- 0.22 and 1.96 +/- 0.08 kPa/mm, respectively. Histological analysis revealed that the viable chondrocytes and neochondrocytes at 12 weeks accounted for 62.0% +/- 3.45%, 65.89% +/- 48% in the autograft group and 60.1% +/- 3.98%, 55.20% +/- 7.57% in the allograft group. No marked immunological differences between the auto- and allograft groups were noted. CONCLUSIONS: Free auricular cartilage can be used to reconstruct the extensive anterior defect of trachea in both auto- and allo transplantations. PMID- 12575207 TI - [Study on the relationship between cervical cancer and p53 codon 72 polymorphism]. AB - OBJECTIVE: A polymorphism at codon 72 of the human tumor-suppressor gene, p53, results in translation to either arginine or proline. Some recent reports suggested that the risk of HPV-associated cervical cancer in white women is higher for those homozygous for the arginine allele than for those who are heterozygous; some reports did not. So we examined a number of cervical cancer, ovarian cancer and normal control to see if we could confirm this result. METHODS: Three groups of women were studied: 20 women for normal control; 15 women with ovarian cancer; 15 women with cervical cancer. DNA from peripheral blood samples and from pathologic tissue sections was examined by PCR with allele specific primers. RESULTS: The proportions of individuals homozygous for the arginine allele, homozygous for the proline allele and heterozygous for the two alleles were 38%, 6% and 56% in normal women; 38%, 5% and 57% in women with ovarian cancer; 78%, 8% and 14% in women with cervical cancer, respectively. Chi square analysis showed significant differences in the proportions. CONCLUSION: In this population, individuals homozygous for the arginine variant of codon 72 of the p53 gene were at increased risk of cervical cancer. PMID- 12575208 TI - [Study on the expression of adhesion molecule related proteins in patients with coronary heart disease]. AB - OBJECTIVE: To assess the expression level of adhesion molecule related proteins CD11a and CD11b on neutrophils, monocytes and lymphocytes in patients with coronary heart disease and detect the relation of expression level to the degree of stenosis of coronary arteries and the severity of coronary heart disease. METHODS: The subjects were divided into three groups: stable angina, unstable angina, and healthy people as control. The amount of cell adhesion molecule related proteins CD11a and CD11b on neutrophilic granulocytes, monocytes and lymphocytes of peripheral blood was measured in three groups by using fluorescent immunoassay method on flow cytometer. Relationship between them was analysed. RESULTS: No significant difference in the expression of CD11a between the angina and control groups was observed. The expression of CD11b on lymphocytes and neutrophilic granulocytes in the unstable angina group and stable angina group was significantly higher than that in the healthy people(P < 0.01 and P < 0.05). Although the amount of CD11b measured in the group of severe coronary artery stenosis was higher than that measured in the mild light group, the difference was not significant (P > 0.05). CONCLUSION: The amount of adhesion molecule related protein CD11b on neutrophilic granulocytes and lymphocytes in patients with unstable angina is higher than that in patients with stable angina and control group, but there is no relationship between the expression level of CD11b and the degree of coronary stenosis. PMID- 12575209 TI - [Relation between insulin resistance and A1166C molecular variant of type 1 angiotensin II receptor gene in patients with coronary heart disease]. AB - OBJECTIVE: To explore the relation between insulin resistance(IR) and A1166C molecular variant of type 1 angiotensin II receptor(AT1R) gene in Chinese subjects with coronary heart disease(CHD). METHODS: IR was calculated by 1/(fast plasma insulin x fast plasma glucose). The A1166C molecular variant of AT1R gene was determined by polymerase chain reaction and restriction endonuclease analysis methods in 90 patients with coronary heart disease and 80 healthy people. RESULTS: The frequency of allele C of group CHD was significantly higher than that of control group. There was no significant difference of IR level among AT1R A1166C AA, AC, CC genotypes in patients with CHD(P > 0.05). CONCLUSION: No significant association between A1166C molecular variant of AT1R gene and insulin resistance was found in Chinese with CHD. PMID- 12575210 TI - [The effect of interferon alone, cytosine arabinoside alone and their combined effect on hematopoietic progenitors of chronic myeloid leukemia in vitro]. AB - OBJECTIVE: In order to search for more effective treatment regimen of chronic myeloid leukemia (CML). METHODS: Semisolid culture and conventional chromosome assay were used to evaluate the effect of interferon (IFN) and Ara-c on K562 cell line and BMMNCs from CML patients and normal controls. RESULTS: IFN had no obvious inhibitory effect on K562 cell. IFN and Ara-c had preferential inhibitory effect on CFU-GM growth of CML and selective cytotoxic effect on CML Ph+ cell. The combination of IFN with Ara-c enhanced the cytotoxic effect of IFN or Ara-c alone on CML cells. CONCLUSION: IFN and Ara-c can be selected for therapy of CML in chronic phase. The combination of IFN with Ara-c may be more effective than IFN or Ara-c alone for the therapy of CML. PMID- 12575211 TI - [Randomized controlled trial for the effect of amrinone and aprotinin on expression leukocyte adhesion molecule in patients with prosthetic valve replacement during perioperative period]. AB - OBJECTIVE: To explore the effect of amrinone and aprotinin on expression of leukocyte adhesion molecule in patients with prosthetic valve replacement during perioperative period. METHODS: Thirty-two patients undergoing valve replacement were randomized to control group, the first aprotinin group, the second aprotinin group, and amrinone combined with aprotinin group; each group consisted of eight cases. In the first aprotinin group, 3 x 10(6) units of aprotinin was added to the priming solution of the extracorporeal circulation (ECC). In the second aprotinin group, 3 x 10(6) units of aprotinin was added to the priming solution of ECC, which was combined with 1 x 10(6) units of aprotinin for intravenous bolus. In the amrinone combined with aprotinin group, 3 x 10(6) units aprotinin was added to the priming solution of the ECC and amrinone began with a bolus of 1 mg/kg followed by a maintenance intrusion of 8 micrograms/kg.min. The control group received an equivalence without aprotinin. Venous blood samples were drawn before the operation, at the end of the ECC, 1 hour after the end of the ECC, and one day after the operation, respectively. Flow cytometry was used to demonstrate the expression of leukocyte adhesion molecule CD11b/CD18. RESULTS: Before ECC, there were no differences of the levels of CD11b+/CD18+ in all groups (P > 0.05). One hour after the ECC, the level of CD11b+/CD18+ in group B was lower than that in group A, the level of CD11b+/CD18+ in group C was lower than that in group B, and the level of CD11b+/CD18+ in group D was higher than that in group C, but no significant difference between groups was noted (P > 0.05). CONCLUSION: Although amrinone and aprotinin have anti-inflammatory activity, the pump prime which was added aprotinin alone or aprotinin combined with amrinone might fail in preventing the expression of leukocyte adhesion molecule CD11b/CD18 completely in patients with prosthetic valve replacement during ECC perioperative period. PMID- 12575212 TI - [Clinical observations on the prevention and treatment of premature labor with nifedipine]. AB - OBJECTIVE: To study the effect of a calcium channel antagonist--Nifedipine(NIF) on the prevention and treatment of premature labor, the suppression of uterine contraction, the prolongation of pregnancy, and to observe the side effects of NIF. METHODS: Eighty-four patients with threatened premature labor were randomly divided into three groups according to the dosage of NIF. Group A received NIF according to a 10 mg dosing schedule; Group B received NIF according to a 20 mg dosing schedule; Group C(not treated with NiF) served as control. The time of delivery delayed, the relationship of therapeutic effect with cervical dilatation and gestational age, and the results of delivery were observed and analyzed. RESULTS: In this study, INF effectively suppressed uterine contraction and delayed the time of delivery. The suppressant intensity was associated with the dosage. The therapeutic effects was apparently more noticeable when the treatment was initiated before the active opening of cervical os. No side effect of NIF was observed in Groups A and B. CONCLUSION: Nifedipine is highly effective as a tocolytic agent for the management of premature delivery. It is of clinical usefulness, and the results of the above observations merit further study. PMID- 12575213 TI - [Multicenter evaluation on the efficacy and safety of sparfloxacin in the treatment of acute bacterial infections]. AB - OBJECTIVE: To evaluate the efficacy and safety of domestic sparfloxacin. METHODS: A multicentre randomized controlled clinical trial was conducted to compare sparfloxacin versus ofloxacin in the treatment of acute bacterial infections. 58 patients received 200 mg of sparfloxacin once daily intravenous drip and 61 patients received 200 mg of ofloxacin twice daily intravenous drip. Both drugs were given for 7-14 days. RESULTS: The clinical cure rates of sparfloxacin group and ofloxacin group were 44.83% and 42.62%, and the efficacy rates were 87.93% and 81.97%, respectively; no significant differences were noted between the two groups (P > 0.05). The bacterial eradication rates were 86.00% and 84.62% respectively (P > 0.05). The adverse reactions in the two groups were 13.79% and 22.58% (P > 0.05). Photosensitivity reaction was not observed in this study. CONCLUSION: Domestic sparfloxacin is effective and safe in treating acute bacterial infections. PMID- 12575214 TI - [Research on preparation of mycobacterium tuberculosis DNA microarray]. AB - OBJECTIVE: To optimize and develop the technique for mycobacterium tuberculosis DNA microarray. METHODS: The process included preparation of DNA samples, spotting and past-spotting treatment of arrays. DNA microarrays were prepared by spotting fluorescence labeled PCR products of target genes onto specially treated glass slides with robotics. The fluorescent signals before and after treatment were scanned with a scanner, and the DNA attachment rate was calculated from the obtained data by software. RESULTS: A foundation for optimizing the conditions of Mycobacterium tuberculosis DNA microarrays has been laid. The support aldehyde modified glass slide is useful for anchoring DNA at Some distance. DMSO as spotting solution is of benefit to preparation of Mycobacterium tuberculosis DNA microarray. Drying the chip at 37 degrees C temperature after spotting can enhance the DNA combination rate. CONCLUSION: Several key steps of this technique have been optimized. This study has provided a foundation for optimizing the DNA attachment conditions in creating mycobacterium tuberculosis DNA microarray. PMID- 12575215 TI - [The primary culture of human tooth pulp cell]. AB - OBJECTIVE: In this study, we established the optimal condition and duration for the primary culture of the human tooth pulp cell with collagenase digestion method. METHOD: Pulp tissue was removed from healthy young human teeth extracted for orthodontic purposes, and then the pulp was digested by collagenase separately in duration of 30 mins, 1 hr, 2 hrs, 3 hrs. After the digestion, the viability and digestion efficiency were evaluated by microscopy and trypan-blue dying. The cytokeratin and vimentin were immunocytochemically detected to identify the cell phenotype. The tissue explant culture and trypsin digestion were set as controls. RESULTS: After digestion of 1 hr, 2 hrs, or 3 hrs, little tissue residue was left, while there is still much tissue remained after digestion of 30 mins. The viability decreased with the elongation of digestion duration. CONCLUSION: 37 degrees C, continuing stirring and 1 hr of type I collagenase digestion are the optimal conditions for the primary culture of human tooth pulp with digestion method. PMID- 12575216 TI - [Study on preparation and tissue distribution of SEA liposomes]. AB - OBJECTIVE: To make a study on the preparation and tissue distribution of staphylococcal enterotoxin A (SEA) liposomes and to provide scientific basis for the therapy of liver cancer by using SEA liposomes. METHODS: SEA liposomes were prepared by reverse-phase evaporation; the diameter and entrapment efficiency (EC) of SEA liposomes were determined. 125I labeled SEA solution and 125I-SEA liposomes were administrated intravenously to mice, respectively. The radioactivity of the organs was determined by gamma-counter. RESULTS: The mean diameter and EC of SEA liposomes were 505 +/- 34 nm and 44.1% +/- 4.8%, respectively. SEA liposomes were found mainly distributed in the liver and spleen. SEA liposomes had a higher blood clearance, compared with SEA solution; SEA solution had high-radio-activity in plasma and kidney; there was statistical significance between the two groups (P < 0.05). CONCLUSION: The preparation method of SEA liposomes is simple and repeatable. SEA liposomes possess liver targeting properties and may provide a new application foreground for the treatment of liver cancer. PMID- 12575217 TI - [Expression of c-terminal of Parkin in E. coli and the preparation of antiserum]. AB - OBJECTIVE: To clone and express 3'-terminal of Parkin gene in E. coli, and prepare its antiserum for further study. METHODS: The glutathion-sulfate transferase (GST) fusion expression plasmid of 3'-terminal of Parkin gene (937 1959 bp) was constructed and transferred to JM 105. After being treated with Triton-100 (1%) and Tween-20 (1%) and purified with affinity chromatograph, GST Parkin C was used to immunize New Zealand rabbits to acquire antiserum. Antiserum was analysed with immunoblot. RESULTS: The GST-Parkin C protein was expressed in JM 105, existing in the form of inclusion body with a molecular weight of around 42 kD; The purity of GST-Parkin C was up to 95%; the titer of antiserum was 1:64; Immunoblotting showed that the prepared antiserum could react specifically with 51.6 kD protein extracted from the mouse brain. CONCLUSION: A high level of expression of GST-Parkin C is obtained in JM 105, and its antiserum can be prepared successfully. PMID- 12575218 TI - [Transarterial intrahepatic xenotransplantation of microencapsulated newborn porcine islets--an experimental study of type 1 diabetes mellitus]. AB - OBJECTIVE: To evaluate the physiologic features and probable side effects of transarterial intrahepatic xenotransplantation of microencapsulated newborn porcine islets. METHODS: Six artificial diabetic canines were transplanted intrahepatically with 6-9 x 10(5) encapsulated newborn porcine islets without immunosuppression. Before and after the transplantation, the serum C peptide, glucose tolerance test, and dosage of insulin were measured. Samples of the livers and pancreases were examined pathologically 6 months after the operation. RESULTS: After the transplantation of microencapsulated porcine islets, the serum C peptides were elevated significantly. The plasma glucose returned to normal gradually. The dosage of insulin decreased. Six months after the operation, the usage of insulin in 2 dogs was stopped and the normal plasma glucose was kept for 180 days. The pathologic examination showed the xenografts located in the interlobular artery of the livers and the livers of canines were not damaged. CONCLUSION: The transplantation may correct the high plasma glucose level of diabetic canines without immunosuppression and has no side reactions. PMID- 12575219 TI - [A case report of cones degeneration]. PMID- 12575220 TI - [Molecular conformation and expression efficiency in the hepatocyte targeting gene drug]. AB - OBJECTIVE: To modulate the molecular conformation of the hepatocyte targeting drug to increase exogenous gene expression efficiency in the targeted cell. METHODS: Some specific auxiliary molecules were added to the gene drug its molecular liquid conformation was investigated under the electron-microscope and they were transfected into human hepatoma cell line BEL-7402 in vitro. We detected IFN-gamma gene expression product by ELISA and screened out the most efficient drug molecules. RESULTS: Due to various concentrations of auxiliary molecules, the conformation of hepatocyte targeting drug changed, including floccule, globulous, stringbeads, bacillform and chromosome-like mixture. While the most efficient expression conformation was the bacilliform and chromosome like mixture and didn't need to inhibit the lysozyme activity, the expression efficiency of such drug molecules was much higher than that of the liposome targeting vector. CONCLUSION: The molecular drug conformation had much influence on targeted gene expression efficiency in gene therapy. Bacilliform and chromosome-like mixture may be the most efficient expression conformation to construct drug molecules in ASOR targeted hepatocyte gene therapy. So we put forward a new concept "chromosome mimic conformation" in screening out the most efficient molecular drug conformation in gene therapy. PMID- 12575221 TI - [Protective effect of Na(+)-H+ exchanger inhibitor cariporide on the injury of vascular endothelial function induced by hypercholesterolemia]. AB - OBJECTIVE: To explore possible effects of cariporide, a new Na(+)-H+ exchanger (NHE-1) inhibitor, against injuries of vascular endothelial function induced by hypercholestrolemia. METHODS: Twenty-two healthy New Zealand rabbits were randomly divided into 3 groups: Six were fed with normal diet as the control; eight were fed with high cholesterol plus high fat diet (HCHF) as the injured control group; and the other eight were fed with HCHF plus cariporide orally (0.1 mg/kg.d-1) as the treated group. The animals were weighed and their blood was drawn from the central ear artery to analyze plasma cholesterol at the pre experiment and in the 10th week of postexperiment. The animals were killed in the 10th week and the aortic artery was isolated to analyze vascular endothelial function. RESULTS: 1. The HCHF diet significantly attenuated endothelium dependent relaxation (EDR) of rabbit aortic rings to acetylcholine (ACh), but cariporide significantly protected against HCHF diet-injured EDR of rabbit aortic rings. The dose-response curves elicited by ACh from HCHF diet rabbits were more significantly shifted to the right than those from HCHF diet plus HOE 642-treated rabbits. 2. The HCHF diet and cariporide did not significantly influence the endothelium-independent relaxation of the rabbit artery elicited by sodium nitroprusside (SNP). 3. There was no significant difference in the levels of total cholesterol and LDL-C between the HCHF diet and the HCHF diet plus cariporide rabbits. CONCLUSION: The Na(+)-H+ exchanger (NHE-1) inhibitor can protect against injuries of vascular endothelial function induced by hypercholesterolemia. The activized Na(+)-H+ exchangers of cellular membranes may be involved in injuries of vascular endothelial function induced by hypercholesterolemia. PMID- 12575222 TI - [Cloning of cDNA fragments of human defensins]. AB - In order to acquire the cDNA of a small molecular peptide with biological activity, the cDNA encoding human defensins was isolated from total RNAs of peripheral leukocyte in a patient with chronic myelogenous leukemia by the RT-PCR method, about 280 bp in length. The RT-PCR products were cloned into the pUCm-T vector, and may be used to construct a novel fusion protein, which contains human defensins and human bactericidal permeability increasing proteins. PMID- 12575223 TI - [Therapeutic effect of recombinant human interleukin-11 on thrombocytopenia models induced by anti-platelet serum in guinea pigs]. AB - OBJECTIVE: To observe the effect of recombinant human interleukin-11 on the platelet counts and the CFU-Meg of the marrow cell culture. METHODS: Thrombocytopenia models of guinea pigs were made by injecting subcutaneously anti platelet serum. RESULTS: 200, 100, and 50 micrograms.kg-1 of rHuIL-11 increased the CFU-Meg of the marrow cell culture and the peripheral platelet, showing a significant difference from that of the control group (P < 0.05). CONCLUSION: rHuIL-11 has a significantly therapeutic effect on thrombocytopenia models induced by anti-platelet serum in guinea pigs. PMID- 12575224 TI - [Detection of staphylococcal clumping factor with purified fibrinogen]. AB - A modified method of detecting staphylococcal clumping factor with purified fibrinogen was introduced. The results showed that the method was simple and concordant with the one with rabbit plasma. Moreover, the results were easy to observe. The traditional method with human plasma was found to be likely to produce false positive. PMID- 12575225 TI - [A case of deep vein thromboembolism and both pulmonary embolism with respiratory failure]. PMID- 12575226 TI - [mRNA expression of transforming growth factor-beta 1 in the cornea after photorefractive keratectomy in rabbits]. AB - OBJECTIVE: To investigate the roles of transforming growth factor beta 1 (TGF beta 1) during healing after photorefractive keratectomy (PRK). METHODS: Twenty of 24 rabbits underwent bilateral 193 nm excimer laser photorefractive keratectomy to correct 10 diopters of myopia, and the other 4 rabbits were taken as the normal group. The mRNA levels of TGF-beta 1 in the corneas of the rabbits were determined by in situ hybridization with a Digoxigenin labled probe. RESULTS: 1. Corneal haze appeared in operated eyes from the 7th day, and peaked on the 28th day after PRK. The most severe haze reached Grade 3. 2. In the normal group, a low level of mRNA expression of TGF-beta 1 was observed in the epithelium, and no positive result in the stroma. On the 7th, 14th, and 28th days after the operation, the mRNA levels of TGF-beta 1 in the epithelium and stroma of the operation group were significantly higher than those in the normal group. CONCLUSION: TGF-beta 1 may involve in corneal wound healing after PRK and may increase haze by promoting the synthesis of the extracellular matrix and the proliferation of keratocytes. PMID- 12575227 TI - [Transforming growth factor-beta 1 expression in epithelial ovarian neoplasms and its clinical significance]. AB - OBJECTIVE: To detect the expression of transforming growth factor-beta 1(TGF-beta 1) in epithelial neoplasms and its clinicopathologic features and prognosis. METHODS: TGF-beta 1 expression was detected in 77 cases of epithelial ovarian neoplasms by SABC immunohistochemistry (including 21 benign tissues, 7 borderline tissues, and 49 epithelial ovarian cancer tissues), and they were compared with 14 normal ovarian tissues. RESULTS: 1. The positive rate of TGF-beta 1 expression was 71.42% in the normal ovary, 76.19% in benign tissues, 4/7 in borderline tissues and 38.78% in epithelial ovarian cancer tissues. 2. TGF-beta 1 expression was associated with the grade, the FIGO stage, and the histologic type of the patients with epithelial ovarian cancer (P < 0.05). CONCLUSION: The expression of TGF-beta 1 may play an important role in the progression of epithelial ovarian cancer and may be related to the prognosis of epithelial ovarian cancers. PMID- 12575228 TI - [Distribution of hepatitis B virus genotype in Hunan Province and its clinical significance]. AB - OBJECTIVE: To study the distribution of hepatitis B virus genotype in Hunan Provine and its clinical significance. METHODS: HBV genotype was determined by the restriction fragment length polymorphism analysis in 185 PCR positive patients, including 42 asymptomatic HBV carriers (ASC), 38 chronic mild or moderate hepatitis (CH), 80 fulminant hepatic failure (FHF), and 25 hepatocellular carcinoma (HCC) patients in Hunan Province. RESULTS: Of the 185 patients, 136 (73.5%) were genotype B, and 49 (26.5%) were genotype C. There was a statistical significance in the distribution of genotype B between FHF and ASC, and between HCC and ASC (83.7% vs. 57.1%, 76% vs. 57.1%, P < 0.01, respectively). Vertical transmission and HBeAg positivity were higher in genotype C than in genotype B (38.8% vs. 13.2%, 57.1% vs. 30.9%, respectively, P < 0.001). The ALT value was significantly higher in genotype B than in genotype C (P < 0.001). CONCLUSION: Genotypes B and C exist in Hunan. Genotypes B is the major genotype in this area and associated with the development of severe liver diseases. Genotype C is associated with vertical transmission. PMID- 12575229 TI - [Liposome and complete Freund's adjuvant for the prevention of insulitis in non obese diabetic mice]. AB - OBJECTIVE: To explore the effects of complete Freund's adjuvant (CFA), incomplete Freund's adjuvant(IFA), large multilamellar liposome (LML) and small cationic liposome (DOTAP) on insulitis and diabetes. METHODS: 1. 3-week-old non-obese diabetic(NOD) female mice were randomly divided into 3 groups: CFA group (injected subcutaneously in the hind footpad, n = 16), IFA group (injected intraperitoneally, n = 16) and PBS group (injected subcutaneously in the hind footpad, n = 16). Three mice from each group (9 in total) were killed at the age of 12 weeks for the analysis of pancreatic pathology, and the others were not killed until they were 30 weeks old for diabetes incidence. 2. 3-week-old NOD female mice were randomly divided into 3 groups and injected intraperitoneally with LML (n = 13), DOTAP (n = 13) and PBS (n = 13), respectively. The insulitis score and diabetes incidence were estimated in the same way. RESULTS: 1. CFA injected subcutaneously in the hind footpad could significantly reduce the insulitis score and decrease diabetes incidence in NOD mice. At the age of 30 weeks, the incidence of diabetes in the CFA group was lower than that in the PBS group (injected subcutaneously in the hind footpad) (9.2% vs 81.8%, P = 0.001). At the age of 12 weeks, the insulitis score in the CFA group was lower than that in the control group [(0.05 +/- 0.02) vs (0.54 +/- 0.11), P < 0.001]. 2. LML injected intraperitoneally could significantly reduce the insulitis score in NOD mice. At the age of 12 weeks, the insulitis score in the LML group was lower than that in the PBS control group (injected intraperitoneally) [(0.16 +/- 0.02) vs (0.58 +/- 0.06), P < 0.001]. At the age of 30 weeks, there were no significant differences in the diabetes incidence between the LML group and the PBS group (60% vs 90%). 3. The protective effect of CFA was better than that of LML in NOD mice. CONCLUSION: CFA injected subcutaneously in the hind footpad may prevent NOD mice from developing diabetes and reduce the insulitis severity. Although the protective effect of CFA is better than that of LML, LML can lessen the insulitis severity and may become a new preventive strategy in NOD mice. PMID- 12575231 TI - [Effects of nao-yi-an granule on bcl-2 expression in the rat brain after experimental cerebral hemorrhage]. AB - OBJECTIVE: To investigate the protective effects and mechanisms of the traditional Chinese medicine complex, nao-yi-an granule (NYA) on experimental intracerebral hemorrhage (ICH) in rats. METHODS: ICH models anesthesized were induced by stereotaxical injection of 2 microliters normal saline containing 0.4 U bacterial collagenase(type VII) into the right globas pallidus(GP), and the protein expression of bcl-2 in the rat brain after ICH was detected with the immunohistochemistry method, with which the effects of NYA were observed. RESULTS: Bcl-2 positive cells were found in the cerebral cortex and basal ganglia area after ICH. Immunoreactivity was localized in the cytoplasm as a fine granular precipitate. The levels of bcl-2 expression in the model group varied at different time-points, and reached the peak at 2 d. In the NYA group, the protein expression of bcl-2 was increased significantly both in the cortex and in the basal ganglia compared with those in the model group. CONCLUSION: NYA can significantly upregulate the protein expression of bcl-2. PMID- 12575230 TI - [Effects of nao-yi-an granule on the expression of heme oxygenase-1 in brains of intracerebral hemorrhagic rats]. AB - OBJECTIVE: To investigate the effects of the traditional Chinese medicine complex nao-yi-an granule(NYA) on the expression of heme oxygenase-1(HO-1) in the brains of intracerebral hemorrhagic (ICH) rats. METHODS: After inducing ICH rat models with collagenase VII, we used the immunohistochemical method and HO-1 immunoreactive cell count to observe the HO-1 expression. RESULTS: Following ICH, the expression of HO-1 in the rat brains was observed at 12 h, peaking at 2 d and persisting until 7 d; and NYA could increase the expression at 24 h obviously. CONCLUSION: Expression of HO-1 increases following ICH,; upregulation of HO-1 expression may be one of the neuroprotective mechanisms of NYA. PMID- 12575232 TI - [Effects of nao-yi-an on tumor necrosis factor alpha and insulin resistance of acute intracerebral hemorrhagic patients]. AB - OBJECTIVE: To investigate the effects of nao-yi-an on tumor necrosis factor alpha (TNF-alpha) and insulin resistance of acute intracerebral hemorrhagic patients. METHODS: The patients were randomly divided into two groups: the medicine control group and the nao-yi-an treatment group. TNF-alpha and insulin were checked by radioimmunoassay before and after the treatment. Fifteen healthy people were made as the negative control group. RESULTS: TNF-alpha of both treatment groups was higher while insulin sensitivity index(ISI) was remarkably lower compared with the health group (P < 0.01). After the treatment ISI increased and TNF-alpha was remarkably lower in the treatment group (P < 0.01). Differences of TNF-alpha and ISI between the treatment group and medicine control group were significant (P < 0.05 or P < 0.01). CONCLUSION: nao-yi-an can decrease TNF-alpha and increase ISI, which is a mechanism of reducing cerebral ischemic-reperfusion injury. PMID- 12575233 TI - [Effect of ischemic preconditioning on human lung cell apoptosis in vivo and the expression of regulating gene bcl-2]. AB - OBJECTIVE: To investigate whether ischemic reperfusion injury induces lung cell apoptosis and the effect of ischemic preconditioning on lung cell apoptosis by altering the expression of bcl-2 protein. METHODS: Sixteen patients, who needed to occlude the main pulmonary artery to undergo pulmonectomy, were randomly put to two groups: the control group (Group C) and the ischemic preconditioning group (Group I). Ischemic preconditioning was achieved by two 5-minute cycles of ischemic, and each was followed by a 5-minute reperfusion. The lung tissue was sampled at the time of preoccluding, 30 minutes after the ischemic, and 30 and 60 minutes after the reperfusion. Apoptotic cells were stained by the terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling (TUNEL) technique. Immunohistochemistry was used to detect the expression of bcl-2 protein. RESULTS: The apoptosis index (AI) of lung cells increased significantly 30 and 60 minutes after the reperfusion, and also significantly increased as the reperfusion period prolonged. Apoptosis was significantly reduced in Group I 30 and 60 minutes after the reperfusion compared with that of Group C at the same time points (P < 0.05). Immunohistochemistry analysis showed that the expression of bcl-2 protein significantly increased in Group I compared with that of Group C(P < 0.05), while there was no significant difference at each time point in Group C(P > 0.05). CONCLUSION: 1. Ischemic reperfusion injury may induce human lung cell apoptosis. 2. Ischemic preconditioning may reduce the apoptosis of human lung cells in vivo by upregulating bcl-2 protein expression. PMID- 12575234 TI - [High frequency color duplex sonography in cervical spondylosis]. AB - OBJECTIVE: To observe the morphological structure and hemodynamic changes of the vertebral artery in cervical spondylosis by high frequency color duplex sonography. METHODS: We measured 138 cases of cervical spondylosis at the vertebral artery with a 7.5-10 MHz probe of high frequency color duplex sonography, to determine morphological changes, Diameter(D), Velocity(V), Blood flow volume, and Resistance index(RI) and (PI). A controlled analysis was performed at the same time. RESULTS: The measurement showed that the vertebral arteries of the patients bent more severely than the controlled group. There was a significant difference between the two groups (P < 0.01) in Diameter (D), Velocity (except mean velocity), Blood flow volume, and resistance index (RI) and (PI). CONCLUSION: High frequency color duplex sonography has provided clinical judement on morphological and blood flow dynamic parameter changes of the vertebral artery with a valuable objective basis. PMID- 12575235 TI - [Mitral valve replacement with a preserved subvalvular apparatus]. AB - OBJECTIVE: To compare the differences of mitral valve replacement between preservation and no preservation of subvalvular apparatuses for severe mitral regurgitations with mitral stenosis. METHODS: We collected and analyzed the data of cardic function obtained from the cardic colour echo six months before and after operation from 41 patients with severe mitral regurgitation and mitral stenosis, 21 of whom had mitral valve replacement with the preservation of a subvalvular apparatus (MVRP), and 20 of whom had mitral valve replacement without the preservation of a subvalvular apparatus (MVR). RESULTS: The internal diameter of the left ventricle, left ventricular end-diastolic volume index (LVEDVI) and left ventricular end-systolic volume index (LVESVI) were remarkably decreased in the group of MVRP as well as MVR, and the rate of the low cardic output syndrome and death obviously decreased. CONCLUSION: MVRP is superior to MVR for severe mitral regurgitation with mitral stenosis. PMID- 12575236 TI - [Electrophoresis and the traditional method for the measurement of lipoprotein cholesterol in serum]. AB - OBJECTIVE: To compare the advantages and disadvantages of electrophoresis with the traditional method in measuring lipoprotein cholesterol in serum. METHODS: Four lipoprotein cholesterols were determined by the Helena electrophoretic system. Quantification of high density lipoprotein-choleterol(HDL-C) was performed by phosphotungstate-magnesium precipitation or direct HDL-C assay. The concentration of low density lipoprotein-cholesterol (LDL-C) was calculated by F equation. RESULTS: Four lipoprotein zones were segregated distinctly by the electrophoretic method. Lp(a) zone appeared only in 6 out of 68 normal samples (8.82%). Reference values of HDL-C, Lp(a)-C, VLDL-C, and LDL-C in serum were 1.16 2.19, 0-0.22, 0.07-0.47, and 1.86-3.46 mmol.L-1, respectively. When comparing the results by electrophoresis with those by the precipitation method for HDL-C, no significant difference was found (P > 0.05); but there was a significant difference between electrophoresis and F-formula calculation for LDL-C (P < 0.05). When comparing results of electrophoresis with the direct method, we found significant differences in both HDL-C and LDL-C (P < 0.01). CONCLUSION: F-formula used for calculating LDL-C is not appropriate to clinical application. Electrophoresis allows the quantification of HDL-C, VLDL-C, LP(a)-C, and LDL-C. The method is simple, convenient, and appropriate to routine application. PMID- 12575237 TI - [Application of three dimensional CT reconstruction in the diagnosis and treatment of bone and joint injury and spinal stenosis]. AB - OBJECTIVE: To explore the application of three dimensional(3D) CT reconstruction(3D-CT-R) in spinal stenosis and injury of the bone and joint. METHODS: Forty cases were examined with the X-ray film, CT scan and 3D-CT-R. To evaluate their diagnostic value, the images of the three methods were compared. RESULTS: X-ray films could show a visible shift of fracture pieces, but we were sometimes incapable of diagnosing fractures accurately from X-ray and, therefore, we couldn't diagnose fractures located at a special position; the CT scan could show the fractured place with a better result, but it required that doctors should have enough clinic experience, and should be familiar with local anatomy to avoid possible misdiagnosis. CONCLUSION: 3D-CT-R could be used in formulating a particular treatment scheme, and is an ideal examination method. PMID- 12575238 TI - [Randomized double-blind clinical trial of moderate dosage naloxone in acute moderate and severe traumatic brain injury]. AB - OBJECTIVE: To evaluate the efficacy and safety of moderate dosage naloxone in acute moderate and severe traumatic brain injury. METHODS: A randomized double blind prospective clinical trial was done to compare the differences of naloxone and saline in acute moderate and severe traumatic brain injury. Naloxone or saline placebo was intravenously given for 10 days. We followed up for at least 1 month. The indexes of assessment of prognosis were Glasgow outcome scale, verbal function and motor function. RESULTS: Forty cases were enrolled in the clinical trial, evenly divided into the naloxone group, and the saline group. On the 10th day after the treatment, Glasgow coma scale, improvement of abnormity of blood pressure, rhythm of the heart and breath in the naloxone group were significantly higher than those in the saline group. The mortalities of the naloxone group and the saline group were 0 and 5% respectively. After the one-month follow-up, Glasgow outcome scale and verbal function in the naloxone group were significantly higher than those in the saline group. In addition, a patient was found with mania possibly caused by naloxone. CONCLUSION: Early application of moderate dosage naloxone in acute traumatic brain injury may significantly reduce the mortality rate and improve the recovery of nerve function. PMID- 12575239 TI - [Pathogen variance and drug-sensibility analysis of lower-respiratory-tract bacterial infection in chronic pulmonary heart disease]. AB - We contrastively analyzed pathogens and their drug sensibilities of lower respiratory-tract infection in chronic pulmonary heart disease in two different periods. The results were that the Gram-positive cocci (GPC) was 43.8% and Gram negative bacilli (GNB) was 56.2% in Group A (1985-1990); GPC was 24.6% and GNB was 75.4% in Group B (1995-2000). The predominant bacteria were in the following order: Preudomonas (21.9%), pneumococcal (18.8%), streptococcus (10.9%), acinetobacter (9.4%), staphylococcus epidermidis (7.8%), and klebsiella (7.8%) in Group A; and preudomonas (30.7%), acinetobacter (12.3%), staphylococcus epidermidis (12.3%), klebsiella (10.5%), enteric bacilli (7.0%), and staphylococcus aureus (7.0%) in Group B. The results suggest that GNB is the main pathogen in recent years; sensibility of most drugs in the 1990s fell in varying degrees compared with that in the 1980s, but antibacterial in common use is still available. PMID- 12575240 TI - [Efficacy of operation in the treatment of cervical spondylotic myelopath in different disease periods]. AB - OBJECTIVE: To analyze the efficacy of cervical spondylotic myelopathc (CSM) in different disease periods after operation, and explore the best operating period for CSM treatment. METHODS: Eighty-nine cases of CSM were included in this study. Thirty-eight cases were treated by posterior open-door or twain-door laminoplasty, and 34 cases were treated by anterior "opening window" decompression and auto-iliac bone graft. According to the disease period, 89 cases were classified into group I (< 6 months), group II (> 6 months, < 12 months), and group III (> 12 months). RESULTS: Seventy-two cases were followed up for an average of 18 months (ranged 10 to 60 months). According to the JOA standand, the statistical results of the postoperative improvement rate of showed that the shorter the course of the disease, the better the surgical result. In patients operated on within 6 months of the disease, the surgical result was the best(P < 0.01), and when the course of disease was beyond 12 months, the operative result was obviously poor (P < 0.01). CONCLUSION: Patients with CSM should undergo operation within 6 months with no contraindication. Even if the disease period is beyond 6 months, patients also should be operated on as soon as possible. PMID- 12575241 TI - [A case report of pulmonary cryptococcosis neoformans]. PMID- 12575242 TI - [Value of simultaneous determination of serum hyaluronic acid, collagen type IV and the laminin level in diagnosing liver fibrosis]. AB - OBJECTIVE: To study the value of simultaneous determination of serum hyaluronic acid(HA), collagen Type IV (CIV) and the laminin (LN) level in the diagnosis of liver fibrosis. METHODS: Serum HA, CIV and LN were measured by ELISA in 22 healthy persons, 10 cases of acute hepatitis, 10 cases of chronic hepatitis, 10 cases of heary chronic hepatitis, and another 25 cases of posthepatitic cirrhosis. The sensitivity, specificity and accuracy of simultaneous determination were compared with those of single determination in posthepatitic cirrhosis. RESULTS: The levels of serum HA, CIV and LN in posthepatitic cirrhosis cases and heary chronic hepatitis cases were significantly higher than those in the other groups (P < 0.01). Simultaneous determination of these indices enhanced the diagnostic sensitivity, but decreased the diagnostic specificity. CONCLUSION: Simultaneous determination of HA, CIV and LN can enhance the diagnostic sensitivity. Simultaneous determination of HA and CIV is an optimal choice. PMID- 12575243 TI - [Antibacterial activity of Chinese medicine kou-kang mouthwash and its acute toxicity test]. AB - OBJECTIVE: To study the antibacterial activity of Chinese medicine kou-Kang mouthwash and its acute toxicity, and provide the basis of experiment and therapy for infectious diseases of the oral cavity. METHODS: Minimal inhibitory concentrations (MIC) of kou-Kang mouthwash were determined by the agar dilution method, and the acute toxicity was done. RESULTS: The range of MIC kou-Kang mouthwash against 27 Gram negative bacterial were 0.03-1.0 g.ml-1. MIC50 was 0.125 g.ml-1 and MIC90 was 0.125-0.5 g.ml-1. The range of MIC kou-Kang mouthwash against 21 Gram positive bacterial was 0.008-0.5 g.ml-1. The range of MIC50 and MIC90 were 0.015-0.125 g.ml-1 and 0.015-0.5 g.ml-1, respectively. The value of MIC was 0.015 g.ml-1 against candidia albicans. The acute toxic test indicated that the maximum tolerance dose was 417.6 g.kg-1 and 210 times as much as the clinical dose a day. No death of animals or toxicity and side-effect occurred in the test. CONCLUSION: Kou-Kang mouthwash shows a potent antibacterial activity against various pathogens of the oral cavity. Its antibiotic activity against Gram positive bacteria was 2-8 times stronger than that against Gram negative bacteria. It has low toxicity and can be used in clinical application. PMID- 12575244 TI - [Surgical treatment of infective endocarditis: a report of 41 cases]. AB - OBJECTIVE: To study the characteristics and surgical treatment of infective endocarditis. METHODS: In all patients, surgical treatment was performed including aortic valve replacement in 22, mitral valve replacement in 9, combined aortic and mitral valve replacements in 4, tricuspid valve reconstruction in 3, and pulmonary valve repair in 3. Meanwhile, complicated deformities such as ventricular septal defect (VSD), atrial septal defect (ASD), patient ductus arterisus (PDA), ruptared aneuryem of the aortic sinus and right centricular outflow tract obstruction were corrected. RESULTS: There were 2 early postoperative deaths (an overall hospital mortality of 5%). A follow-up of 3 months to 5 years, with a mean of 3.2 years, documented no recurrent endocarditis and late death. CONCLUSION: Low mortality occurs in the surgical therapy for infective endocarditis. In order to avoid irreversible injury on cardiomyocyte, the operation, which is beneficial to the recovery of heart function, should be performed as early as possible. PMID- 12575245 TI - [Clinical analysis of 228 patients treated with penctoneal dialysis]. AB - OBJECTIVE: To summarize the treatment with peritoneal dialysis with the "O" set peritoneal dialysis catheter. METHODS: Reviewing the 228 cases of chronic renal failure patients, we compared their blood pressure and hemoglobin before peritoneal dialysis with those after peritoneal dialysis, observed the morbidity of peritonitis and drainage block, and analyzed reasons that the patients withdrew from peritoneal dialysis. RESULTS: The patients' blood pressures were controlled very well after peritoneal dialysis. Hemoglobin increased significantly, compared with that before peritoneal dialysis. Death caused by complications of peritoneal dialysis was the primary reason that the patients withdrew from peritoneal dialysis. CONCLUSION: Peritoneal dialysis is simple, convenient, safe and efficient. PMID- 12575246 TI - [Extended-spectrum beta-lactamase detection in Enterobacteriaceae and antibiotic susceptibility analysis]. AB - OBJECTIVE: To detect the extended-spectrum beta-lactamases (ESBLs) in family Enterobacteriaceae and analyze the antibiotic susceptibility of those ESBLs producing strains. METHODS: ESBLs were determined by the double-disk confirmatory test and 8 antibiotic susceptibilities were tested with the disk disffusion method in those strains producing ESBLs. RESULTS: Forty-seven ESBLs-producing strains comprised of 25 of E. coli, 14 of K. pneumoniae, 5 of E. cloacae, 1 of K. oxytoca, 1 of K. rhinoscleromatis, and 1 of S. liquefaciens. The susceptibility rates of those strains were: 100% for imipenem and meropenem, 89.4% for piperacillin/tazobactam, 72.4% for cefoxitin and 65.9% for cefotetan. CONCLUSION: E. coli and K. pneumoniae are the prime strains producing ESBLs in Enterobacteriaceae. Imipenem and meropenem are the best drugs to deal with those ESBLs-producing strains. Piperacillin/tazobactam is better than cephamycins and other beta-lactama/beta-lactamase inhibitor combination. PMID- 12575247 TI - [Anterior transfer of the gracilis in repairing of recurrent dislocation and subluxation of the patella]. AB - OBJECTIVE: To introduce anterior transfer of the gracilis in the treatment of recurrent subluxation and dislocation of the patella, and evaluate its effects. METHODS: Six patients with recurrent subluxation of the patella and 2 patients with postoperative recurrence of dislocation of the patella were treated with anterior transfer of the gracilis based on the lateral retinacular release and medial tightening. The efficacy was reviewed. RESULTS: The mean follow-up time was 22.4 months. No recurrence of subluxation of the kneecap occurred, and 7 cases had excellent results. CONCLUSION: Anterior transfer of the graciclis in repairing recurrent subluxation of the patella is easy to perform, and can effectively avoid the recurrence of subluxation of the patella. PMID- 12575248 TI - [Psychological health status and nursing countermeasures of cerebral arterial sclerosis patients]. AB - OBJECTIVE: To estimate the psychological health status of attacking stage patients with cerebral arterial sclerosis(CAS). METHODS: Sixty in-patients were surveyed by filling up the questionnaire (SCL-90) and compared with the customary model in China. RESULTS: The symptomatic factor scores of somatization and depression of the patients were obviously higher than those of the customary model (P < 0.001). Their symptomatic factor scores of anxiety and obsessive compulsive were higher than those of the customary model (P < 0.05). The symptomatic factor score of phobic anxiety was higher than that of the customary model (P < 0.01). CONCLUSION: Somatization, obsessive-compulsive, depression, anxiety and phobic anxiety are common psychological problems. Corresponding nursing countermeasures are proposed in our study. PMID- 12575249 TI - [Simultaneous determination of four components in the compound child phenobarbital tablet using ultraviolet spectrophotometry]. AB - OBJECTIVE: To establish a method to determine four components in the child phenobarbital tablet. METHODS: Ultraviolet spectrophotometry was used to determine four components without separation. RESULTS: The contents of aspirin, phenacetin, caffeine and phenobarbital could be measured simultaneously. The average recoveries of four components in simulated samples and the compound child phenobarbital tablet samples were 99.0%, 98.9%, 99.8%, and 101%, respectively, and relative standard deviations of those were 2.0%, 2.0%, 2.9%, and 2.2%, respectively. CONCLUSION: The method is simple, fast, reliable, and can be used to monitor the quality of compound drugs. PMID- 12575251 TI - [Determination of lipopolysaccharide by capillary zone electrophoresis]. AB - OBJECTIVE: To detect lipopolysaccharides (LPS) in frangibility bacilli, control the procedure of LPS preparation, examine the ally of the product by high performance capillary zone electrophoresis (CZE), and obtain rapidly high alloy of LPS for scientific research. METHODS: Theoptyline was added to the sample as an internal standard. The LPS was separated and detected by CZE in sodium tetraborate-borate buffer. The concentration of the buffer was 30 mmol.L-1 (pH 8.0); the diameter of the capillary column was 75 microns, and the length was 57 cm; the CZE operative voltage was 25 kV, the electric current was 112 microA, and the detecting wavelength was 230 nm. RESULTS: CZE was first internally used to separate and detecte LPS in bacillus. The LPS had a good linearity in the range of 2-30 mg.L-1 (r = 0.996). The average recovery rate was 99%-101%, the in-day and day-to-day coefficients of variation (CV) were less than 5%, and the minimum detectable quantity was 1.0 mg.L-1. CONCLUSION: Compared with the limulus test (LT) and gel electrophoresis, this method can resist heat source interference and is more simple, rapid and accurate. Moreover, it can control the preparative process of LPS immediately, and may be an advanced method to determine LPS. PMID- 12575250 TI - [Isolation and purification of islets in non-obese diabetic mice]. AB - OBJECTIVE: To investigate the method of isolating and purifying islets in non obese diabetic (NOD) mice. METHODS: The isolation of adult NOD mice pancreatic islets was carried out by stationary digestion after pancreatic ductal injection of collagenase solution. A discontinuous Ficoll solution (25%, 23%, 20.5%, 11%) was applied to the purification of the islets. The specificity of the islets was determined by dithizone(DTZ) staining, and the function was evaluated by the glucose stimulating insulin releasing test. RESULTS: The yield and activity of the islets varied with different concentration collagenase and digesting times. After incubation of the pancreas with 1.0 mg.ml-1 Type XI collagenase for 40 min, each pancreas could yield (195 +/- 8) islets, with viability > 90% and purity > 90%. The islets were red and intact after DTZ staining. The insulin from the islets fed with high concentration glucose was 2 times that with low concentration. CONCLUSION: The modified method of pancreatic ductal collagenase injection, followed by stationary digestion and purification of islets with a discontinuous Ficoll solution, can reproducibly yield various and pure islets. PMID- 12575252 TI - [Polymorphism of the HLA-DMA in Han nationality in Hunan Province]. PMID- 12575253 TI - [Effect on characterization of Ca3 (PO4)2 ultra-fine powder by sintering temperature]. PMID- 12575254 TI - [Researching some sensitivity factor of bioassay method on the antibiotics pharmacokinetics]. PMID- 12575255 TI - [Resources and distribution of Cordyceps sinensis in Naqu Tibet]. AB - This paper reported the resources and distribution of Cordyceps sinensis in Naqu district, the principally grown zone in Tibet. The investigation showed that the yearly yield of Cordyceps sinensis are 7000 kg, the distribution areas are Baqin, Suoxian, Biru, Jiali, Naqu and Nierong county at 30 degrees 20'-32 degrees 48' N, 92 degrees 0'-94 degrees 50' E. The rangs of vertical distribution from the elevation 5000 m to 4100 m; the ecological geographical distribution is affected mainly by vegetation, soil, temperature and humidity. PMID- 12575256 TI - [The descriptions, microscopic and TLC identification of Tribulus terrestris L]. AB - The descriptions, microscopic and TLC identification of Tribulus terrestris L. were carried out. It provided evidences for the identification of Tribulus terrestris L. PMID- 12575257 TI - [Study on processing conditions of wine baked radix Achyranthis bidentatae]. AB - According to the content of oleanolic acid in Radix Achyranthis Bidentatae, the conditions of wine baking were studied with orthogonal test. The results showed that optimum conditions are using 20% yellow wine, baking at 120 degrees C for one hour. PMID- 12575258 TI - [Analysis of the constituents of essential oil from radix Polygoni multiflori preparata]. AB - The chemical constituents of oil from Radix Polygoni Multiflori Preparata were analyzed by GC-MS-DS.27 constituents were identified. PMID- 12575259 TI - [Study of chemical constituents of essential oil from Michelia yunnanensis Franch]. AB - The essential oil from Michelia yunnanensis Franch were extracted and chemical constituents of it were analyzed by GC/MS. Forty-five compounds were identified. The main compounds with high contents were Bornyl acetate (12.44%), Camphor (8.775), Caryophylle oxide (6.93%), Bornyl (5.97%), Cineole (4.57%) and Linalool(1.89%). PMID- 12575260 TI - [Experimental studies on antioxidation of extracts from several plants used as both medicines and foods in vitro]. AB - OBJECTIVE: The scavenging effects of 8 extracts from several plants used as both medicines and foods on oxygen free radicals were studied in vitro. METHODS: Chemiluminescence (CL) method and ultra-weak luminescence technique were used, meanwhile, macrophage respiratiory burst was also carried out to evaluate the antioxidation reaction. RESULTS: The scavenging activity on superoxide anions (O2 .) differed in the following order: extract of clove (CE) > black tea (BTE) > green tea(GTE) > grape peel/seed (GE) > or = oologn tea (OTE) > > tumeric (TE). In Fenton reaction for hydroxyl radicals (.OH), the order as follows: BTE > CE > or = GTE > GE > OTE > > TE. Most extracts have stronger eliminating efficiency on .OH than on O2-. except CE, which may scavenge the O2-. formation more efficiently. After macrophages (5 x 10(6) cells/ml) were stimulated with phorbol myristate acetate, BTE showed stronger inhibiting action than CE on respiratory burst in macrophages. The results of ultra-weak luminescence measurement indicated that the extracts could delay and depress the DNA damage in CuSO4-Phen Vc-H2O2-DNA chemiluminescence system. CONCLUSIONS: It suggests that the tested antioxidants could decrase the formation of O2-. or .OH, some have chain-breaking function and preventative function during DNA damage. PMID- 12575261 TI - [Effect of qiongyugao on the action of DDP in inhibiting the division of GLC-82 cell strain in vitro]. AB - OBJECTIVE: To observe the effect of Qiongyugao (QYG) on the action of DDP in inhibiting the division of GLC-82 cell strain in vitro. METHODS: The method of serum pharmacology was used in observing the morphological change of the cancer cells. By accounting the number of the live cells of each day during 6 days respectively, the growth curve was drawn up. The effects of the three groups on the cancer cells apoptosis of GLC-82 cell strain were analysed by FACS. RESULLTS: The growth of the cancer cells in control group (normal saline) was much better than the other two groups; in chemical therapy group (DDP), the number of the live cells almost kept steady; the number of the live cells in combined group(DDP plus QYG) became even lower. QYG could enhance the action of DDP in inducing the apoptosis of GLC-82 cells strain remarkably. CONCLUSION: QYG can enhance the effects of chemotherapy in inhibiting cancer cells division and inducing their apoptosis. PMID- 12575262 TI - [The study on Rhizoma Alpiniae officinarum and other herbs as penetration enhancer for the permeation of 5-fluorouacil]. AB - OBJECTIVE: To study the effects of volatile oils from Rhizoma Alpiniae Officinarum, Pericarpium Zanthoxyli, Herba Asari, cineaol and ethanol extracts on the percutaneous pentration of 5-fluorouracil. METHODS: By Valia-Chienhorizontal diffusion cell and HPLC, the effects of volatile oils from Rhizoma Alpiniae Officinarum, Pericarpium Zanthoxyli, Herba Asari, cineol and ethanol extracts on the percutaneous penetration of 5-fluorouracil were observed and compared with azone. RESULT: 1%, 3% volatile oils from Rhizoma Alpiniae Officinarum, Pericarpium Zanthoxyli and cineol possessed strong promoting action on percutaneous absorption of 5-fluorouracil. The effects of volatile oils from Rhizoma Alpiniae Officinarum was stronger than that of azone; the effects of volatile oils from Pericarpium Zanthoxyli was the same as that of azone. CONDUSION: The volatile oils from Rhizoma Alpiniae Officinarum, Pericarpium Zanthoxyli, Herba Asari can enhance effectively the skin permeation of 5 fluorouracil. They deserved to be researched and developed further. PMID- 12575263 TI - [A herbalogical study on traditional Mongolian medicine dakesha]. AB - By herbological study and investigation, "Dekesha" used by Mongolia doctors mainly contains 17 species from 3 genus of 3 family, but "Dakesha" in Inner Mongolia Standard of Medicinal Materials only contains 2 species which are Oxytropis myriophylla and O. hirta. PMID- 12575264 TI - [Effects of cultivating measures on root rot of Panax notoginseng]. AB - OBJECTIVE: To probe into the effects of cultivating measures on root rot of Panax notoginseng for the integrate control. METHOD: Field investigation on the planting plots was conducted. CONCLUSIONS: Effects of cultural system on root rot are obvious, (1) The rotation period should be more than eight years; (2) Improper fertilizer application is one important fact of the root rot occuring; (3) The light transmitting rate is one important fact; (4) Laying film and using sun-shade shed can reduce the root rot. PMID- 12575265 TI - [Numerical taxonomy of medicinal plants from Areae in Zhejiang Province]. AB - Numerical taxonomic studies were done on 13 species from 3 genera of tribe Areae in Zhejiang, used as 13 operational taxonomic units (OTUS). 40 morphological characters were used for analysis. Euclid distance coefficients used to show quantitative index of similarity among OTUS were computed by standardized data. The dendrograms from 8 systematic cluster methods including single linkage method, complete linkage method, median method (beta = 0 WPGMA) and (beta = -0.25 WPGMA), centroid method, group average method, variable group average method (beta = -0.25) and variable method (beta = 0.25) were constructed respectively. WPGMA was slected as the optimal one by computing the comparative coefficients of every cluster result and the boundary of taxa in its dendrogram determined by a method to treated IBM computer with the program by BASIC language. The Results showed that 13 OTUs were classified as 3 clusters by broken line L2, namely, cluster I: Typhonium; cluster II: Arisaema; cluster III: Pinellia, and cluster II further as 4 groups by L1, which belong to Section Pistillata, Arisaema, Sinarisaema and Tortuosa respectively. Being consistent with those from classical taxonomy, the results in this study may be helpful to the classification and identification of the medicinal plants from tribe Areae in Zhejiang Privince. PMID- 12575266 TI - [The identification of Bungarus multicinctus multicinctus and its confused species by SDS-PAGE]. AB - The comparative study of the proteins of Bungarus multicinctus multicinctus and Natrix annularis was carried out by SDS-PAGE. The results showed that there were significant differences between their SDS-PAGE figures. On the basis of the study the molecular weights of their characteristic proteins were determined. The results could be regarded as a reference for identification of Bungarus multicinctus multicinctus and its confused species. PMID- 12575267 TI - [Studies on chemical components of the volatile oil from the leaves of Hypericum perforatum]. AB - OBJECTIVE: To determine the volatile oil of the leaves from Hypericum perforatum L.. METHOD: The essential oil was obtained by simultaneous distillation and solvent extraction. The chemical components were seperated and identified by GC MS. The relative contents in the volatile oil were determined by area nomalization. RESULTS: 54 chemical constituents were separated and identified representing 92.32% of the total contents. CONCLUSION: Sesquiterpenes are major chemical constituents in the oil from the leaves of Hypericum perforatum L.. They are very different from those of the essential oil from the same species from the other country. PMID- 12575268 TI - [Content analysis on gardenoside in grown and ungrown fruits of Gardenia from different habitats]. AB - This paper reported content analysis on gardenoside in grown fruits of Gardenia from different habitats and ungrown fruits of Gardenia jasminoides and G. jasminoides f. longicarpa. The results were as follows: (1) The content of gardenoside in G. jasminoides from indigenous region of Jiangxi was higher than other habitats; (2) The content of gardenoside in G. jasminoides f. longicarpa was the highest; (3) The content of gardenoside in fruits of growth period of G. jasminoides and G. jasminoides f. longicarpa had two peak stages. PMID- 12575269 TI - [Studies on the antimutagenic effect of asparagus juice]. AB - Micronucleus (MN) test, sister chromatid exchanges (SCE) and sperm malformation test were used to study the effects of asparagus juice on the mutagenic activity of cyclophosphamide (CP). It was found that, asparagus juice itself showed no effcet on the incidence of MN, SCE and sperm malformation, but asparagus juice reduced the incidence of MN (from 60.0@1000 to 50.3@1000-26.6@1000), SCE (from 29.22 times/cell to 26.40-13.37 times/cell) and sperm malformation (from 61.0@1000 to 51.2@1000-28.6@1000) induced by CP respectively. The inhibitory effects showed a dose dependent relation, which was statistically significant. PMID- 12575270 TI - [Preliminary screen of anti-endotoxin activity of Chinese materia medica]. AB - OBJECTIVE: To screen Chinese materia medica against E. coli. O111B4 endotoxin. METHOD: 70% alcohol extracts from 134 kinds of heat-clearing Chinese materia medica were demonstrated by using limulus test in vitro. The extracts which had anti-endotoxin effects were diluted to demonstrate their minimal anti-endotoxin concertnations. RESULTS: 75 kinds of Chinese materia medica possessed anti endotoxin activity in various degrees in vitro. CONCLUSION: Most of heat-clearing Chinese materia medica possess anti-endotoxin effects in vitro. PMID- 12575271 TI - [Effects of an extract of Ginkgo biloba on the blood flow of brains and back legs of dogs]. AB - After injecting the injection of an extract of Ginkgo biloba, the cerebral blood flow of dogs wrer increased (P < 0.05-0.001). So were the back legs. And the cerebrovascular resistance was decreased (P < 0.05-0.001). PMID- 12575272 TI - [Study on the technological process of including volatile oil of Baizhu with beta cyclodextrin]. AB - The volatile oil of Baizhu was included with beta-CD. The factors affecting the formation of the inclusion compounds were researched by L9(3(4)) orthogonal test. The ratio of the oil in inclusion compounds and recovery of inclusion compounds were used as criteria in the test. The best conditions for inclusion were A2B3C1, that is, the compounds should be formed in the aqueous solution containing beta CD and oil with a raio of 8:1, aqua destillate and beta-CD with a ratio of 4:1 for two hours. PMID- 12575273 TI - [Chemical constituents of grub and grub eye drops]. AB - The chemical constituents of grub (Holotrichia diomphalia Bates.) were systematically analyzed. The results showed that grab contained amino acids, polypeptide or protein, carbohydrate, alkaloid, organic acid, steroid et al. The analysis of amino acids in the hydrolysate in grub eye drops showed the content of Glu and Gly was high, which occupied 40.33% of total amino acids in the hydrolysate. We inferred that Glu and Gly might be the active principle on catatract and nebula. PMID- 12575274 TI - [Preliminary report on introducing exogenous DNA into Panax quinquefolium]. AB - Worked aimed at introduction of Panax ginseng's exogenous DNA into Panax quinquefolium has been done continuously the past three years in our institute. The results showed that the best way of exogenous DNA introduction is via the pollen tube similarly lesser flowery organ. The suitable introducing time is at 10:00-11:00 am, and the seeds ratio is 13.21% at the experiment limits. PMID- 12575275 TI - [The ejection of ascoporae of Cordyceps sinensis]. AB - This paper has reported the effect of sunshine to stromata of Cordyceps senensis grown in Kanding, Sichuan and Naqu, Tibet. The ejective time, quantity and germination rate of ascoporaes between the two habitats are different with sunshine. To satisfy the needs of sunshine, moisure and temperature can be raised the ejection and germination of ascorporae. PMID- 12575276 TI - [Pharmacognostical identification of Phyllanthus niruni]. AB - The article reported the Chinese medicational materias descriptions, the microscopic characteristics and the physical-chemical identification of zhuzicao (Phyllanthus niruni Linn). PMID- 12575277 TI - [Morphological and histological identification and essential oil GC-MS assay of Aralia decaisneana]. AB - This paper reported the studies on morphological and histological diagnostic characteristics and powder feature the root of Aralia decaisneana Hance. Chemical analysis indicates essential oil from root cortex contains at least 16 constituents which trans-caryophllene, 9,12-octadecaienoic acid, etc. are the major ingredients. PMID- 12575278 TI - [Isolation and identification of a new long-chain tetrahydroxyamide]. AB - A novel long chain tetrahydroxyamide compound (C42H85NO5, mp 178-180 degrees C) has been isolated from Polymastia sobustia collected from the South China Sea. On the basic of the spectral data of IR, MS, 1H NMR, 13C NMR, DEPT and chemical way, the compound was identified as N-(1-hydroxymethyl-2,6-dihydroxyl) heptadecyl-5 hydroxyltetracosanamide, sobustiamide. PMID- 12575279 TI - [Study on quality evaluation of Forsythia suspensa with grey pattern recognition]. AB - In this paper, based on the grey relative relation grade, a new pattern recognition model for quality evaluation of traditional Chinese medicine was presented. As an example, the model was used to evaluate the quality of Forsythia suspensa, the results are satisfactory. PMID- 12575280 TI - [GC-MS analysis of chemical components in essential oil from Flos magnoliae]. AB - In this paper, the chemical components and their relative contents of essential oil in three kinds of Xinyi(Magnolia biondii, Magnolia denudata and Magnolia sprengeri) were identified and analyzed by GC-MS. The main components are 1,8 cineole, sabinene, beta-pinene, alpha-pinene, trans-caryophyllene, etc. PMID- 12575281 TI - [The preventive effect of the oil from the seed of Torreya grandis cv. merrillii on experimental atherosclerosis in rats]. AB - OBJECTIVE: To determine the preventive effect of the oil from the seed of Torreya grandis cv. merrillii (OTM) on experimental atherosclerosis in rats. METHODS: The concentration of blood lipids, plasma thromboxane (TXA2), prostacyclin (PGI2), the ratio of TXA2/PGI2 and endothelin (ET) were investigated in male Wistar rats fed high-fat feed (HFF) containing OTM(2%) for 12 weeks. RESULTS: OTM could reduce the level of serum total cholesterol (TC), triglyceride (TG) and the atherosclerosic index (AI), and increase the level of serum high-density lipoprotein cholesterol (HDL-C). The plasma concentration of TXA2, ET and the ratio of TXA2/PGI2 in group supplemented with OTM were lower than those in HFF group. Moreover, the level of plasma PGI2, was higher than that in HFF group. CONCLUSION: OTM is beneficial in preventing atherosclerosis. PMID- 12575282 TI - [Experimental studies of icariin on anticancer mechanism]. AB - Icariin (ICA) is a kind of new biological respone modifier(BRM) and differentiational agent. In order to further elucidate the reversion of malignant phenotypes of tumor cells and the mechanism of its action, highly metastatic human lung cancer cells(PG) were treated with ICA in vitro. In this study, MTT assay, radioimmune assay, flow cytometry(FCM) and invasion assay were used. The results showed that ICA could influence the distribution of PG cells cycle and reduce S phase. Moreover, ICA increased the level of cAMP in PG cells, reduced the level of cGMP and increased the cAMP/cGMP ratio. On the other hand, ICA decreased PG cells adhesive ratio to laminin substrate and decreased the ability of invasion or migration. These data demonstrate that ICA maybe a kind of effective anticancer drug. PMID- 12575283 TI - [Study on activating blood and eliminating stasis of total paeony glycoside(TPG)]. AB - The effects of Total Paeony Glycoside(TPG) on activating blood and eliminating stasis were studied. The results showed that TPG could significantly improve the microcirculation, reduce the specific viscosity of serum and plasma, inhibit the aggregation of platelet induced by ADP and prolong PT and KPTT. PMID- 12575284 TI - [Experimental study on decoctum Agrimonia pilosa Ledeb-induced apoptosis in HL-60 cells in vitro]. AB - In this study, MTT assay, cell morphology and flow cytometry were used to explore Agrimonia pilosa Ledeb on HL-60 cells in vitro. The data suggested that Agrimonia pilosa Ledeb had the effects of growth inhibition in HL-60 cells. After treated with decoctum Agrimonia pilosa Ledeb, cells underwent apoptosis with typical morphological feature. DNA histograms by flow cytometric analysis displayed distinct apoptotic peak. PMID- 12575285 TI - [Determination of glycyrrihizin in rats plasma and pharmacokinetics after oral administration of si-jun-zi-tang]. AB - OBJECTIVE: To study the pharmacokinetics of glycyrrhizin after oral administration of Si-jun-zi-tang in rats and provide the experimental evidences for the prescription pharmacokientics of traditional Chinese medicine. METHOD: A RP-HPLC method was established for rapid determination of glycyrrhizin in rats plasma. After obtaining the blood sample at different time, the plasma concentration of GL were detected respectively. The data were processed by the software named 3P87 to find the pharmacokinetic parameters. RESULTS: A good linear relationship existed in the range of GL from 1.224 micrograms/ml to 38.656 micrograms/ml. Whole blood concentration-time course of GL after oral administration Si-jun-zi-tang was fitted to be a two-compartment model. GL in rats plasma which came from Si-jun-zi-tang was absorbed and distributed rapidly, but excreted slowly. CONCLUSION: This RP-HPLC method was sensitive, raliable and rapid, it can be used in absorption and metabolism of GL, also in detection of GL preparation. This experiment is prepared for further study of the tradtional Chinese medicine prescription pharmacokinetics. PMID- 12575286 TI - [Expression and purification of p11 fusion protein in E. coli and preparation of antiserum against GST-p11]. AB - OBJECTIVE: To obtain p11 fusion protein and prepare specific polyclonal antibody against p11. METHODS: A full-length human p11 gene was cloned into expression vectors, pGEX-4T-2 and pQE30, and transformed into E. coli. The expressed proteins were purified from lysates with Glutathione Sepharose 4B and the Ni-NTA agarose column, respectively. The purified GST-p11 was mixed with Freund's complete or incomplete adjuvant and immunized rabbits. RESULTS: A high level of expression of target proteins was detected after IPTG induction and purified proteins were obtained by affinity chromatography with Glutathione Sepharose 4B and the Ni-NTA agarose column, respectively. Western blotting analysis suggested that the polyclonal antibody can recognize 6xHis-p11 and GST protein. CONCLUSION: The antiserum against p11 prepared by prokaryotic expression of GST-p11 fusion protein has good specificity. PMID- 12575287 TI - [Stimulation of ozone stress on the adhesion of inflammatory cells to bronchial epithelial cells]. AB - OBJECTIVE: To determine into the inflammation process of bronchial epithelial cells (BECs) and observe the modulatory effect of some protective factors on BECs. METHODS: We built the cell injury model with ozone and observed the effect of ozone stress on polymorphonuclear loeukocyte (PMNs) and eosinophil (EOS) adhesion to BECs. The effect of intercellular adhesion molecule (ICAM-1) antibody on the adhesion and ICAM-1 protein expression of BECs was observed at the same time. RESULTS: O3 increased the adhesion of BECs to PMNs and EOS. Adhesion could be blocked by ICAM-1 antibody, suggesting ICAM-1 was the principal molecule leading PMNs and EOS adhesion to BECs. The immunocytochemistry assay indicated that O3 increased the expression of ICAM-1 protein of BECs. CONCLUSION: Ozone stress leads to inflammation, and the principal adhesion molecule is ICAM-1 resulting in adhesion between BECs and PMNs, and between BECs and EOS. PMID- 12575288 TI - [Study of microsatellite polymorphisms between the Han nationality and French population by fluorescence-based genotype]. AB - OBJECTIVE: To facilitate the application of the large scale genome scan and individual identification by investigating the differences of microsatellite polymorphisms of the genethon human genetic linkage map between the Han nationality and French population. METHODS: Six to seventeen pairs of primers were added in one single tube with 5 microliters reaction volume and several microsatellite markers were amplified simultaneously. The products were electrophoresited on the 377XL DNA sequencer. RESULTS: During the entire experiment 400 microsatellite loci were amplified in 32 tubes and 306 microsatellite loci were satisfactory. CONCLUSION: The size range of genetic polymorphisms are different in race, and the difference exists at 124 microsatellite loci between the Han nationality and French population, so 10 bp should be added to both sides of the genethor map size range when we perform genotype in Han nationality. PMID- 12575289 TI - [Killing effect of schistosomula of Schistosoma japonicum by tissues/organs of Microtus fortis in vitro]. AB - OBJECTIVE: To screen the specific tissues/organs of Microtus fortis (MF) that have the activity of anti-schistosomula of Schistosoma japonicum. METHODS: The homogenate supernatant of the heart, liver, lung, spleen, kidney, muscle and fresh serum of MF and those of mice were mixed with schistosomula respectively and cultured at 37 degrees C with 5% CO2. The schistosomula-killing effect of Schistosoma japonicum was observed. RESULTS: There was no significant difference between the tissues/organs of MF and those of the mice in schistosomula-killing effect (P > 0.05), as it was the same with the tissues/organs of MF (P > 0.05) and between the fresh serum of MF and its complement-inactivated serum (P > 0.05). The schistosomula-killing effect of the serum of MF was stronger than that of the mice (P = 0.001), and the same compared to the tissues/organs of MF (P < 0.05). Between the 40% fresh serum of MF and its 20% fresh serum, there was a significant difference (P < 0.001). CONCLUSION: The schistosomula-killing effect of the serum of MF was stronger than that of mice, and it was the same with the tissues/organs of MF. PMID- 12575290 TI - [Comparison of different lysis buffers in the assessment of the efficiency of single cell polymerase chain reaction for preimplantation genetic diagnosis]. AB - OBJECTIVE: To set up a highly efficient cell lysis system to increase the efficiency of single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD). METHODS: First we picked up one single lymphocyte or blastomere and lysed it in alkaline lysis buffer (ALB), proteinase K buffer (PK) and a new proteinase K lysis buffer compounded by ourselves. Then we amplified the 3rd exon of Gene ALD using nested PCR and compared the amplification efficiencies. RESULTS: The amplification efficiencies of the single lymphocyte in ALB, PK and the lysis buffer compounded by ourselves were 90%, 74%, and 87%; those of the single blastomere were 98%, 77%, and 96%. CONCLUSION: ALB and the lysis buffer compounded by ourselves are superior to PK in the lysis of cells in clinical PGD. PMID- 12575291 TI - [High glucose impaires glucose transport and PKB activity in adipocytes of rats]. AB - OBJECTIVE: To explore the effect of high glucose on glucose transport activity, protein kinase B (PKB) activity and glucose transporter 4 (GLUT4) in primary cultured rat adipocytes. METHODS: Isolated rat adipocytes were cultured for 24 h at different glucose concentrations (5, 10, 15 and 25 mmol.L-1). The glucose uptake, cellular and membrane GLUT4 expression, PKB protein expression, and PKB serine phosphorylation and activity were measured. RESULTS: These adipocytes treated with glucose of different concentrations showed that high glucose impaired glucose uptake, PKB phosphorylation and activity, and up-regulated GLUT4 translocation, but didn't affect protein expression of PKB and GLUT4. CONCLUSION: High glucose can induce insulin resistance; the mechanism may be involved in the effect of high glucose on PKB serine phosphorylation and activity as well as GLUT4 function. PMID- 12575292 TI - [Effect of the defect of integrin alpha II b beta 3 on the inside-out signal transduction in platelets]. AB - OBJECTIVE: To investigate the effect of the defect of integrin alpha II b beta 3 on the inside-out signal transduction in platelets. METHODS: The transfected cDNA, its expression and the ability of cells binding to PAC-1 and fibrinogen were investigated by RT-PCR, DNA sequence analysis, flow cytometry and Western blotting. RESULTS: The integrin alpha II b beta 3 level in the patients with Glanzmann's thrombasthenia was significantly lower than that of the normal subjects and the platelets of the patients failed to bind PAC-1 activated by ADP. There were both C3077G and G3078C mutations in exon 30 of mutant alpha II bR995A beta 3 cDNA, which resulted in an amino acid substitution arginine (R) 995 to alanine (A). CHO cells transfected with wild-type alpha II b beta 3 or mutant alpha II bR995A beta 3 cDNA respectively expressed normal alpha II b beta 3 and mutant alpha II bR995A beta 3. When cells were not activated, wild-type alpha II b beta 3 CHO cells failed to bind PAC-1, but could adhere to fibrinogen, but mutant chimera alpha II bR995A beta 3 CHO cells were able to bind PAC-1 and exhibited high affinity binding fibrinogen. CONCLUSION: The point mutation in integrin alpha II bR995A can induce the inside-out signal transduction in platelets, and have integrin alpha II bR995A beta 3 locked into an activation state; the defect of alpha II b beta 3 in patients with GT impairs the inside-out signal transduction mediated by alpha II b beta 3 in platelets and the adhesive functions of platelets. PMID- 12575293 TI - [Proliferation and apoptosis of lung smooth muscular cells in hypoxic pulmonary hypertension in neonatal pigs]. AB - OBJECTIVE: To study the effects of the proliferation and apoptosis of smooth muscular cells (SMC) in small pulmonary arteries on the pulmonary vascular remodeling in hypoxia-induced pulmonary hypertensive neonatal pigs. METHODS: Forty-two pigs aged from the birthday to 14 days were divided into the normal developmental group and hypoxic hypertensive group. Twenty of them were exposed to hypobaric hypoxia (50.8 kPa) for 3 or 11 days to establish a pulmonary hypertension model. The proliferation and apoptosis of SMC in the pulmonary small arteries were studied with the immunohistochemiscal method and terminal transferase-mediated nick-end labeling (TUNEL) method. RESULTS: SMC replication rates were high (5%) at birth, and followed by a burst of replication during the early period after birth. The shorter period hypoxia did not cause the increase of SMC proliferation; and the longer hypoxia induced the pulmonary vascular wall remodeling with apparent SMC proliferation. No signal of cell apoptosis in the pulmonary vascular wall was found during the neonatal period under either the normal condition or the hypoxic condition. CONCLUSION: The pulmonary resistant vascular remodeling after birth is related to SMC proliferation and SMC proliferation plays a crucial role in the formation of severe pulmonary hypertension induced by hypoxia. The role of apoptosis in the pulmonary vascular remodeling needs further research. PMID- 12575294 TI - [Improvement in the dot immunogold filtration assay]. AB - OBJECTIVE: To improve the filtrating unit of the dot immunogold filtration assay (DIGFA) to reduce its cost and make it suitable for fieldwork. METHODS: A thick round filtrating paper disc was made to replace the plastic filtrating box, and their results were compared. RESULTS: The size of the self-made filtrating paper disc was smaller and the cost was lower. The efficacy of antibody examination by our paper disc was the same as that of the usual plastic filtrating box. CONCLUSION: In some aspects, the round filtrating disc is better than the plastic filtrating box. PMID- 12575295 TI - [Transperitoneal transport of solute and IL-8 in dialysis-related peritonitis]. AB - OBJECTIVE: To study the changes in solute transperitoneal transport and the IL-8 level at different time phases in dialysis-related peritonitis so as to understand the mechanism of solute transperitoneal transport. METHODS: Forty-two New Zealand white rabbits were randomly divided into three experimental groups (5 min, 24 h, and 48 h groups) and a control group. The rabbit model of peritoneal dialysis-related peritonitis was created. The plasma and effluent concentration of creatinine, glucose, total proteins, and albumin were determined respectively; the D/P values of creatinine, total proteins, and albumin, and the Dn/D0 values of glucose were calculated respectively. RESULTS: There was a significant difference in the D/P values of albumin at 120, 240, and 360 min between the 48 h group and the control group (P < 0.05); the D/P values of total proteins significantly increased at different time points in the 48 h group compared with the control group (P < 0.05); there was a significant difference in the creatinine D/P values and glucose Dn/D0 values between the 48 h group and the control group (P < 0.05). Compared with the control group, IL-8 and WBC obviously increased in the 48 h group (P < 0.05); the effluent IL-8 levels at different time points were positively correlated to the D/P values of the total proteins (P < 0.05). The effluent IL-8 levels at different time points after 60 min were positively correlated to the D/P values of albumin (P < 0.05). CONCLUSION: The mechanism of transperitoneal transport of the different molecule solutes varies. Both IL-8 and WBC can influence solute transperitoneal transport in peritoneal dialysis-related peritonitis. PMID- 12575296 TI - [Quantitation of telomerase activity using the polymerase chain reaction-based enzyme immunoassay and its clinical application]. AB - OBJECTIVE: To investigate detective methodology of telomerase activity and its clinical application. METHODS: The telomerase activity was quantitated by polymerase chain reaction-base enzyme immunoassay (PCR-EIA). The TS primer was biotinylated (TS-B), and CX primer was digoxigeninated (CX-D). The amplicons containing TS-B were combined with microtiter plate coated streptavidin, then combined with anti-digoxigenin antibody labeled with POD and finally reacted to tetramethylbenzidine substrate solution. RESULTS: Telomerase activity measured by the PCR-EIA method was comparable to that obtained from TRAP-silver stain protocol. The CV of the PCR-EIA method was 4.138%. Telomerase activity was detected in 90% of various tumer tissues. In control tissues, telomerase activity was detected only in 7.1%. CONCLUSION: The PCR-EIA method offers a rapid, quantitative, and nonisotopic assay for the determination of telomerase activity. It is simpler than silver stain protocol. The detection of telomerase activity may play a significant role in the diagnosis of clinical tumors. PMID- 12575297 TI - [Effect of low-dosage aspirin combined with perindopril on prostacyclin, thromboxone A2, and norepinephrine in rabbits' blood]. AB - OBJECTIVE: To explore the interaction of low-dosage aspirin combined with angiotensin-converting enzyme (ACE) inhibitors by prostacyclin (PGI2), thromboxone A2 (TXA2) and norepinephrine (NE)) levels in rabbits' blood. METHODS: Forty healthy New Zealand rabbits were divided into four groups. Blood samples were drawn from the rabbits' heart before and after a consecutive four-week. NE was measured by high performance liquid chromatography, and PGI2 and TXA2 were measured by radioimmunoassay. RESULTS: ACE inhibitors increased PGI2 levels (P < 0.05, P < 0.01); low-dosage aspirin suppressed TXA2 productions (P < 0.05, P < 0.01) after the four-week administration. Aspirin combined with ACE inhibitors led to a significant increase in PGI2/TXA2(P < 0.01), together with a significant decrease in NE levels in the rabbits' blood (P < 0.001). CONCLUSION: Neither low dosage aspirin nor ACE inhibitors influence NE levels alone. The ratio of PGI2 to TXA2 increased, and NE levels decreased significantly during the administration of aspirin combined with ACE inhibitors. The results suggest that there is a synergis-action between low-dosage aspirin and ACE inhibitors due to increased PGI2/TXA2 and decreased NE levels. PMID- 12575298 TI - [Pathological analysis of metastatic brain tumors misdiagnosed with MRI]. AB - OBJECTIVE: To study the pathological basis of metastatic brain tumors misdiagnosed with (MRI). METHODS: Seven cases were analyzed histopathologically and immunohistochemically, compared with the MRI image. RESULTS: All the tumors were single large noduls. Hemorrhage and necrosis of the tumors were seen in all the cases. Cystformation of the tumor, glial cell proliferation and vascular proliferation of peritumoral brain tissues were seen in most cases. CONCLUSION: The secondary changes of metastatic brain tumors and the reactive changes of peritumoral brain tissues are the major causes of MRI misdiagnosis. PMID- 12575299 TI - [Expression of TGF beta 1 and TGF beta RI proteins in tissues of salivary adenoid cystic carcinoma]. AB - OBJECTIVE: To determine the possible role of TGF beta 1 and TGF beta RI in facilitating neoplastic progression in human salivary adenoid cystic carcinoma (ACC). METHODS: The expression of TGF beta 1 and TGF beta RI protein in human salivary adenoid cystic carcinoma was detected by the streptavidin-peroxidase (S P) immunohistochemical method, and their contents were compared with those of normal salivary gland tissues and salivary pleomorphic adenoma (PA). RESULTS: 1. TGF beta 1 and TGF beta RI proteins were expressed in most ductal epithelial cells and myo-epithelial cells, but they were not expressed in the acinar epithelial cells of any normal glands examined; 2. TGF beta 1 and TGF beta RI proteins were expressed in the duct-like arranging tumor cells and other solid proliferating tumor cells of salivary pleomorphic adenoma, and no difference of their contents existed between PA and normal salivary gland tissues (P > 0.05); 3. The contents of TGF beta 1 in ACC were significantly higher than those in normal salivary glands (P < 0.01), while the contents of TGF beta RI in ACC were significantly lower than those in normal salivary glands (P < 0.01). CONCLUSION: The abnormal expression of TGF beta 1 and TGF beta RI might play an important role in tumor genesis and the development of malignant salivary gland tumors such as ACC; the over-expression of TGF beta 1 and low-expression of TGF beta RI might involve the bad biological behavior of ACC. PMID- 12575300 TI - [Experimental research on drug evaluation of ibuprofen oral drop]. AB - OBJECTIVE: To make a pharmaceutical comparison between ibuprofen oral drop and ibuprofen tablet. METHODS: Using ibuprofen oral drop, we observed the anti inflammatory and analgesic effects of ibuprofen oral drop on the inflammatory swelling of the rats' toe caused by diagonal carrageenin; its anti-thermic effect on the temperature rising of domestic rabbits caused by endotoxin; and its analgesic effect on mice's pain caused by acetic acid. RESULTS: Ibuprofen had obvious anti-inflammatory and analgesic functions. CONCLUSION: Ibuprofen oral drop has the same pesticide effect as that of ibuprofen tablet. PMID- 12575301 TI - [Clinical curative effect of dengzhanhua injection on acute cerebral infarction: a report of 100 cases]. AB - OBJECTIVE: To observe the clinical curative effect of dengzhanhua injection on acute cerebral infarction. METHODS: One hundred cases of acute cerebral infarction were treated with dengzhanhua injection (Group A) and 62 treated with fufangdanshen injection (Group B). The curative effects and the patients' hemorrheology and lipemia before and after the treatment were compared. RESULTS: The total effective rate and evidently effective rate of Group A were significantly higher than those of Group B (92.0% vs. 75.8%, 44.0 vs. 29.0%, P < 0.01). The levels of hemorrheology and lipemia in 72 hyperviscosityemia patients and 76 hyperlipidemia patients in Group A were significantly improved after the treatment. The integral difference of nerve function injury and the obsorbed situation of infarct focus in Group A were significantly higher than those in Group B. Group A had no adverse reaction during the treatment. CONCLUSION: Dengzhanhua injection in the treatment of acute cerebral infarction is safe and effective. PMID- 12575302 TI - [Clinical study of amalgam restoration with resin multi-purpose adhesive agent]. PMID- 12575303 TI - [Further study on the indexes of auxiliary laboratory diagnosis in the syndrome of hyperactivity of liver-yang]. AB - The indexes of auxiliary laboratory diagnosis, including plasma norepinephine (NE) levels, plasma epinephrine (E) levels, mean blood flow velocity of the middle cerebral artery (MCA-Vm) and systolic blood flow velocity of the middle cerebral artery (MCA-Vs), were observed in patients with the syndrome of hyperactivity liver-yang. The results indicated that the levels of the 4 indexes were significantly higher in the patients with the syndrome of hyperactivity liver-yang than those in the controls. In hypertension patients with the syndrome of hyperactivity liver-yang, there was a positive correlation between symptomatic scores of the syndrome of hyperactivity liver-yang and diastolic pressure (Pd), plasma NE and E levels. The symptoms ameliorated, and the levels of the 4 indexes decreased correspondingly in 3 weeks after the treatment of Qianyangfang (a traditional Chinese herb). PMID- 12575304 TI - [Hematopoietic stem cell transplantation for hematological malignancies]. AB - OBJECTIVE: To evaluate the efficacy of hematopoietic stem cell transplantation (HSCT) in hematological malignancies. METHODS: Twenty-five patients with hematological malignancies were treated by autologous bone marrow transplantation (ABMT), autologous peripheral blood stem cell transplantation (APB-SCT) or Allo PBSCT. Eight acute myeloid leuekmia (AML) patients who received autologous hematopoietic stem cell transplantation (AHSCT) were treated by chemotherapy again. RESULTS: All the patients engrafted gained hematopoietic reconstitution; they were followed up for 6 to 91 months; five patients died; twenty patients were alive in a disease-free situation. CONCLUSION: After the patients with acute leukemia receive AHSCT, the relapse incidences can be reduced obviously by regular chemotherapy. HSCT is one of the best methods for hematological malignancies. PMID- 12575305 TI - [Clinical value of spiral CT in the surgical treatment of severe acute pancreatitis]. AB - OBJECTIVE: To evaluate the clinical value of spiral CT in the surgical treatment of severe acute pancreatitis (SAP). METHODS: The morphological changes in the spiral CT images of 82 SAP patients were contrasted with the therapeutic methods and the curative effects. RESULTS: The operation for obstructive biliogenic pancreatitis, invasive parapancreas vessels forming angioma, piece and segmental necrosis in the body and tail of the pancreas, the enlargement of pancreas cyst during the treatment, and parapancreas abscess diagnosed by the spiral CT images had a significant curative effect compared with the non-operation treatment (P < 0.05). The satisfactory effect could be gained if a conservative treatment was performed first, and then the delayed operative treatment was carried out according to the etiologic and clinical data for the SAP patients with extensive phlegmona, necrosis in the head of the pancreas, severe parapancreas cyst, and a large amount of ascite. CONCLUSION: Spiral CT can supply the grounds for choosing surgical methods and the surgical opportunity in the treatment of SAP patients. PMID- 12575306 TI - [Electrophoresis in the quantitative determination of cholesterol in lipoprotein fractions and its clinical application]. AB - OBJECTIVE: To evaluate the clinical application of the electrophoresis method for measuring serum cholesterol in lipoproteins and to study the characteristics of lipoprotein fractions in patients with the acute coronary syndrome. METHODS: Cholesterol in lipoprotein fractions in fasting serum was measured by electrophoresis in 26 patients with the acute coronary syndrome and 20 patients with stable angina pectoris. The results were compared to cholesterol in lipoproteins measured with the regular method. RESULTS: There was a good correlation between the methods with coefficients of 0.922, 0.909, and 0.899 for LDL-C, HDL-C and VLDL-C, respectively. The serum levels of TG and VLDL-C in the patients with the acute coronary syndrome were significantly higher than those with stable angina pectoris [(0.47 +/- 0.33) mmol.L-1 vs. (0.29 +/- 0.19) mmol.L 1, P < 0.01, (1.82 +/- 0.70) mmol.L-1 vs. (1.31 +/- 0.48) mmol.L-1, P < 0.01]. CONCLUSION: The electrophoresis method for quantifying cholesterol in lipoprotein fractions is well correlated with the regular method. The serum levels of VLDL-C and TG in patients with the acute conronary syndrome are higher than those with stable angina. PMID- 12575307 TI - [Correlation between blood pressure variability and left ventricular hypertrophy in patients with primary hypertension]. AB - OBJECTIVE: To study the correlation between blood pressure variability (BPV) and left ventricular hypertrophy (LVH) in patients with primary hypertension. METHODS: We monitored the 24-hour ambulatory blood pressure and determined the left ventricular mass index (LVMI) by two dimensional echocardiography in 87 patients with primary hypertension (PH) and 26 normal controls. RESULTS: The LVMI in the PH patients was significantly higher than that in the controls. It was positively correlated to the average level of the 24-hour systolic blood pressure (24 h-SBP), nighttime systolic blood pressure (n-SBP), and nighttime diastolic blood pressure (n-DBP) (r = 0.183, P < 0.05; r = 0.275, P < 0.01; r = 0.243, P < 0.05, respectively). It was also negatively correlated to the 24-hour systolic blood pressure variability (24 h-SBPV) and the daytime systolic blood pressure variability (d-SBPV) (r = -0.253, P < 0.01; r = -0.186, P < 0.05, respectively). The 24 h-SBPV in the patients with LVH was significantly lower than that of those without LVH, but there was no significant difference in d-SBPV and n-SBPV between the PH patients with LVH and those without LVH. CONCLUSION: The 24 h-SBPV is valuable in predicting LVH in hypertension. PMID- 12575308 TI - [Clinical value of hypertonic glucose injection in the treatment of recurrent pneumothorax]. AB - OBJECTIVE: To investigate the clinical value of the 50% hypertonic glucose injection in the treatment of recurrent pneumothorax. METHODS: Forty-five patients with recurrent pneumothorax were injected with 60-80 ml of 50% hypertonic glucose into the pleural cavity after air discharge and then were instructed to turn their body to the right and the left quickly. RESULTS: The curative rate was 100% and the curative effect was satisfactory. There was no other complication except for slight stimulative chest pain in 7 patients. The recurrent rate was 4.4% in a 2-7 year follow-up. CONCLUSION: Treating recurrent pneumothorax with the 50% hypertonic glucose injection is safe, effective, and less recurrent. PMID- 12575309 TI - [A case of ventricular bigeminy with electrical alternans of ventricular premature beat]. PMID- 12575310 TI - [Influence of postprandial hypertriglyceridemia on the endothelial function in elderly patients with coronary heart disease]. AB - OBJECTIVE: To explore the changes of carotid atherosclerosis and the endothelial function after a high-fat meal in elderly patients with coronary artery disease (CAD). METHODS: The carotid intima-media thickness (IMTc) and the brachial artery flow-mediated vasodilation (FMD) before and after a high-fat meal were measured by the high-resolution ultrasound technique in 20 elderly patients with CAD and 22 healthy elderly subjects. RESULTS: The CAD patients had a higher IMTc (P < 0.01) and impaired FMD (P < 0.05) compared with the healthy subjects. The serum triglyceride level increased in the two groups after the meal. The CAD patients had a higher triglyceride peak response (TGPR) than the healthy subjects (P < 0.05). The IMTc of the two groups had no change before and after the meal. The postprandial FMD significantly aggravated in the two groups (P < 0.001); and the percent decrement of FMD in the CAD patients was obviously higher than that in the healthy subjects (P < 0.05). The postprandial TGPR was positively correlated with the fasting IMTc and the postprandial percent decrement of FMD (P < 0.01). CONCLUSION: The disordered postprandial metabolism of triglyceride-riched lipoproteins may play an atherogenic role by inducing endothelial dysfunction. PMID- 12575311 TI - [Clinical efficacy of 0.2% brimonidine in patients with open-angle glaucoma or ocular hypertension]. AB - OBJECTIVE: To evaluate the efficacy and safety of 0.2% brimonidine administered in patients with glaucoma or ocular hypertension. METHODS: Twenty-five patients (41 eyes) with primary open-angle glaucoma or ocular hypertension were enrolled in this prospective study. Eye drops of 0.2% brimonidine were used twice daily for 6 months and the patients were examined every month after the treatment, including the trough intraocular pressure (IOP, IOP 12 h after 0.2% brimonidine was administrated) and the peak IOP (IOP 12 h after 0.2% brimonidine was administrated). The visual field was examined every three months. RESULTS: The use of 0.2% brimonidine significantly lowered the trough IOP and the peak IOP (P < 0.05). Every point of the IOP diurnal curve significantly decreased compared with the baseline (P < 0.05). The visual field had no significant change 3 months or 6 months after the treatment with 0.2% brimonidine. No serious systemic side effect was observed. CONCLUSION: Brimonidine is safe and effective in lowering IOP in glaucoma or ocular hypertension and is well tolerated. PMID- 12575312 TI - [Surgical treatment of congenital heart disease complicated with infective endocarditis]. AB - OBJECTIVE: To review the experience of surgical treatment and the diagnosis of 41 patients with congenital heart diseases complicated with infective endocarditis. METHODS: Twenty-three males and 18 females (the mean age 22.6 years) were studied. Twenty-seven patients received bacterial cultivation of blood specimens, and the results of bacterial cultivation of 11 patients were positive. Thirty four patients underwent cardiac operations after infection was almost controlled, and 7 patients underwent operations during the infective phase. In all the cases, the infected tissues were excised, congenital cardiac was treated, and valvular dysfunctions were corrected. RESULTS: The surgical treatment results were satisfactory. One patient (2.4%) died during hospitalization. CONCLUSION: In our hospital, the incidence rate of infective endocarditis complicated with congenital heart diseases increases; echocardiography is a valuable tool in diagnosis; blood culture may be negative after previous administration of antibiotics; aggressive surgical treatment should be adopted in patients with congenital heart defects complicated with infective endocarditis. PMID- 12575313 TI - [Surgical treatment for pigmented villonodular synovitis]. AB - OBJECTIVE: Pigmented villonodular synovitis(PVS) is a synovial proliferative disorder that remains a diagnostic difficulty. In this retrospective study we evaluated the diagnostic procedures, clinical symptoms and surgical treatment of PVS. METHODS: Six surgically treated cases of PVS were evaluated: five of the lesions were located in the knee joint and one in the hip joint. Diagnosis of PVS was confirmed by histological examination. Operation showed characteristic findings of a joint effusion, and synovial proliferation. RESULTS: Five cases were treated with complete open total synovectomy, and one case received arthoplasty. The followed-up period averaged 3 years and six months (rangeing from 6 months to 6 years). Excellent and good function was seen in all and there was no recurrence. CONCLUSION: PVS diagnosis is frequently delayed due to nonspecific symptoms. Proper treatment includes surgery: extension synovectomy for the diffuse form, and arthoplasty and total joint replacement for the severe bone involvement. PMID- 12575314 TI - [Genetic features of EEG abnormality in children with Tourette syndrome]. AB - OBJECTIVE: To investigate the biogenetic significance of EEG abnormality in Children with Tourette syndrome (TS) and their parents. METHODS: EEG monitoring was carried out for 2 hours in 25 children with TS and their parents. RESULTS: The rate of EEG abnormality was significantly higher than the incidence of tic disorders in parents of TS children. The rate of abnormal EEG in parents of TS children with abnormal EEG was significantly higher than that in parents of TS children with normal EEG. EEG abnormality had no significant differences in parents of males and those of females. CONCLUSION: EEG examination further indicates that Tourette syndrome has an inherit able tendency. PMID- 12575315 TI - [Clinical analysis of 9 cases of the normal-sized ovary carcinoma syndrome]. AB - OBJECTIVE: To investigate clinical features, diagnostic criteria, treatment methods of a clinically rare "normal-sized ovary carcinoma syndrome". METHODS: Histologic slices of 9 cases of the normal-sized ovary carcinoma syndrome from 1990 to 1996 were retrospectively reviewed. Six of the 9 cases were extraovarian peritoneal serous papillary carcinoma(EPSPC). The other 3 cases were serous adenocarcinoma of the ovary (2 cases) and metastatic adenocarcinoma of unknown origin (1 case). All the patients received optimal cytoreductive surgery combined with multiple courses of effective adjuvant chemotherapy. RESULTS: Two patients with EPSPC survived for 84 months and 72 months respectively up to Dec., 1997. The average survival time of the 7 patients who died of their diseases was 16.5 months. CONCLUSION: For the normal-sized ovary carcinoma syndrome, surgical resection is the first choice of treatment, and postoperative chemotherapy is essential to obtain better prognosis. PMID- 12575316 TI - [Influence of 89Sr on the cell immune function in patients with multiple bone metastases]. AB - OBJECTIVE: To investigate the influence of 89Sr on the cell immune function in patients with multiple bone metastases. METHODS: Patients with multiple bone metastases were treated with in vivo radiation of 89Sr. The T cell subsets, NK cell activities, and lymphocyte transformation rate (LTR) of multiple bone metastases before and after the 89Sr treatment were measured. RESULTS: With the palliation of the pain of the patients after the 89Sr treatment, the T cell subset level, NK cell activities, and LTR increased. There was no significant change in the patients with the pain. CONCLUSION: Treating multiple bone metastases with 89Sr can improve the cell immune function of the patients to a certain extent. PMID- 12575317 TI - [Application of rCBF imaging in PD, AD and OPCA]. AB - OBJECTIVE: To evaluate the diagnosis and differential diagnosis of rCBF imaging in Parkison's disease (PD), Alzheimer's disease (AD) and olivopontocerebellar atrophy (OPCA). METHODS: The characteristics of the radioactive distribution of rCBF imaging in PD, AD and OPCA were analyzed, and the laws of the radioactive distribution were identified. RESULTS: The sensitivity of rCBF imaging in PD, AD and OPCA was 88.4% (38/43), 95.8% (23/24), and 85.7% (30/35) respectively. The characteristics of the radioactive distribution were that the radioactivity decreased in the cerebral cortex (25/43), basal nuclei (21/43) and thalamus (18/43) in PD, that the radioactivity decreased in the bilateral cerebral cortex (17/24) in AD, that the radioactivity decreased in the cerebral cortex (17/35), cerebellum (14/35), basal nuclei (14/35) and thalamus (10/35) in OPCA. CONCLUSION: rCBF imaging could sensitively diagnose PD, AD and OPCA, and also could differentially diagnose them according to their different characteristics of the radioactive distribution. PMID- 12575318 TI - [A case of paragonimiasis with spontaneous pneumothorax]. PMID- 12575319 TI - [Detection of telomerase activity in salivary tumor and its significance]. AB - OBJECTIVE: To investigate telomerase activity in human salivary cancer and corresponding adjacent tissues and to explore the possibility of telomerase as a tumor marker and its clinical significance. METHODS: Twenty-eight salivary cancers, 28 adjacent peritumoral tissues, 10 mixed tumors, 6 adenolymphomas, and 5 normal salivary tissues were examined for telomerase activity by the silver staining TRAP assay based on PCR. RESULTS: Twenty-five of the 28 salivary cancers and 2 of the 28 adjacent peritumoral tissue specimens were positive for telomerase activity with a positive rate of 89.3% and 6.3%, respectively. Telomerase activity was negative in the 10 mixed tumors, 6 adenolymphomas, and 5 normal salivary tissues. There was no correlation between the clinical stage of salivary cancer and its expression of telomerase activity (P > 0.05). CONCLUSION: Positive telomerase activity occurs in the majority of salivary cancers examined. It can be used as a tumor marker in the diagnosis of salivary cancer. Detection of telomerase activity in the adjacent peritumoral tissues can be used as a monitoring marker after treatment. PMID- 12575320 TI - [Effect of nicotine on the secretion of TNF of human peripheral blood mononuclear cells in vitro]. AB - OBJECTIVE: To observe the effect of nicotine on the secretion of TNF of human peripheral blood mononuclear cells (PBMC) in vitro, and explore the possible mechanism of the high level of TNF caused by smoking. METHODS: Twenty-one non smokers and 19 smokers were studied. The PBMCs isolated from each volunteer's blood sample were distributed to three groups, and cultured for 72 hours in supplemented RMPI 1640 alone or in supplemented RPMI 1640 containing 50 ng.ml-1 nicotine or 500 ng.ml-1 nicotine respectively. The level of TNF in supernate was quantified with the immuno-radioassay, and PBMC proliferation was observed. RESULTS: The level of TNF spontaneously secreted by PBMCs in smokers was significantly higher than that in non-smokers (P < 0.05). When the concentrations of nicotine were 50 ng.ml-1 and 500 ng.ml-1, the level of TNF increased significantly in non-smokers (P < 0.05, P < 0.01). In smokers, the level of TNF significantly increased when the concentration of nicotine was 50 ng.ml-1; however, the level of TNF significantly decreased (P < 0.05), and the proliferation of PBMCs was inhibited when the concentration of nicotine was 500 ng.ml-1 (P < 0.05, P < 0.05). The level of TNF was positively correlated to the multiple of PBMC proliferation (r = 0.93, P < 0.05). The linear regression equation was Y = 2.913X - 2.955. CONCLUSION: Nicotine can induce PBMCs to secrete more TNF, and the magnitude of the effect is strongly related to the dosage of nicotine, but a great dose of nicotine will inhibit the production of TNF. PMID- 12575321 TI - [Testicular tumors in infants and young children]. PMID- 12575322 TI - [New techniques of postoperative cholelith endoscopy via the "T" tube fistula]. AB - OBJECTIVE: to explore some new techniques of postoperative cholelith endoscopy via the fistula. METHODS: Netting, washing, clamping, pushing, and electrohydraulic techniques were used in 260 cases of postoperative cholelith endoscopy via the fistula and the clinical data were analyzed retrospectively. RESULTS: The stones in 257 cases (98.8%) were removed completely in an average of 2.7 times. Three patients were operated on again because of biliary stricture and huge stones. CONCLUSION: The application of some new techniques in postoperative cholelith endoscopy via the fistula favors the complete removal of stones. PMID- 12575323 TI - [Analysis of amino acids and trace elements in lophatherum gracile brongn and its decoction of water]. PMID- 12575324 TI - [Establishment of regulable cell line for the inducible expression of p53 gene]. PMID- 12575325 TI - [Serum copper and zinc levels of liver disease]. PMID- 12575326 TI - [Value of the high-width and thin coronal CT in sinus surgery]. PMID- 12575327 TI - [Primary characterization and sequence analysis of anti-colorectal cancer phage fusion antibodies]. AB - OBJECTIVE: To identify 5 phage fusion antibodies against colorectal cancer from in vitro immunized phage library and analyze their sequences. METHODS: Cell ELISA, immunohistochemistry, DNA sequencing and computer analysis were employed. RESULTS: Five clones of phage antibodies were tested by cell ELISA, and all of them reacted to human colorectal cancer cell lines, human embryo kidney endothelial cell line and some tumor cell lines, but not to mouse-original cell lines. They also reacted weakly to human hepatic cell lines. The binding specificity of the phage antibodies for colorectal cancer cells was confirmed by immunohistochemistry with cultured cells and colorectal carcinoma and colon tissue sections. They reacted to colorectal carcinoma cell lines, human embryo kidney endothelial cell lines and nasopharyngeal carcinoma cell lines. CH273 reacted specifically to colorectal cancer cells in human colorectal carcinoma sections but not to any of the cells in human colon sections. The 5 clones were further analyzed after their DNA sequencing. The sequences of CH723, CH209 and CHA12 were identical. The lengths of CH273, CH205 and CH723 were 732 bp, 366 bp and 723 bp, respectively. The VDJ regions of CH273, CH205 and CH723 belonged to VH3-30-D1-26-JH3-linker-V1-13-JL2, VH1-46-D6-13-JH3 and VH3-30-D1-26-JH3-linker L2-J kappa 2, respectively. CONCLUSIONS: Phage antibodies binding to colorectal tissues and cells are confirmed, on which human anti-tumor ScFv and VH fragments may be further developed and applied to clinical therapy. PMID- 12575328 TI - [Effects of vasoactive intestinal peptide on the enzyme activity of CTP: phosphorylcholine cytidylyltransferase from the rat lung]. AB - OBJECTIVE: To investigate the effect of vasoactive intestinal peptide(VIP) on the microsomal CTP: phosphorylcholine cytidylyltransferase activity of cultured lung explants and its mechanisms. METHODS: The CDP-[M-14 C] choline content reflecting the microsomal CTP: phosphorylcholine cytidylyltransferase activity of cultured lung explants was measured with liquid scintillation. RESULTS: 1. VIP increased the microsomal CTP: phosphorylcholine cytidylyltransferase activity of cultured lung explants in a dose-dependence manner and in a time-dependence fashion; 2. The effect of 3 x 10(-7) mol.L-1 VIP on the microsomal CTP: phosphorylcholine cytidylyltransferase activity of cultured lung explants was inhibited by H-7, a protein kinase C(PKC) inhibitor and W-7, a calmodulin antagonist respectively. CONCLUSION: VIP in the lung may be one of the key regulators involved in the synthesis of phosphatidylcholine mediated by PKC and calmodulin. PMID- 12575329 TI - [Fractionation and identification of Schistosoma japonicum adult worm 67kD protein]. AB - OBJECTIVE: To isolate the specific protein bands which react to sera from rabbits infected with Schistosoma japonicum (Sj) and those immunized with Sj adult worm antigens (AWA) for immunodiagnosis of schistosomiasis. METHODS: AWA was analysed using SDS-PAGE and immunoblotting, and 67 kD protein was fractionated by electrophoretic chromatography. RESULTS: 67 kD protein which reacted to rabbit sera infected with Sj and immunized with AWA was obtained after the fractionation of AWA through electrophoretic chromatography. CONCLUSIONS: Electrophoretic chromatography is a useful tool for the fractionation of schistosome antigens. PMID- 12575330 TI - [Complete Freund's adjuvant-induced splenic T cell apoptosis in the prevention of diabetes in non-obese diabetic mice]. AB - OBJECTIVE: To study the apoptosis of T lymphocytes in the spleen and its role in complete Freund's adjuvant (CFA) preventing diabetes in non-obese diabetic (NOD) mice. METHODS: Forty-two NOD mice were divided into two groups randomly: CFA treated NOD female mice (6w, 12w and 30w) and PBS-treated NOD female mice (6w, 12w and 30w). The apoptosis of splenic T lymphocytes was observed with the TDT mediated fluorescein-dUTP nick-end labelling (TUNEL) technique and avidin-biotin complex method. RESULTS: CFA induced the apoptosis of CD4+ T lymphocytes in the mouse spleen and could prevent the development of Type 1 diabetes (T1DM) in NOD female mice. CONCLUSION: CFA preventing Type 1 diabetes in NOD mice may be related to the apoptosis of T lymphocytes in the spleen, which downregulates mature CD4+ T lymphocytes in the periphery. PMID- 12575331 TI - [Anti-leukemia effect of sophora flavescens combined with the low molecular weight natural tumor suppressor of the human fetal liver and its mechanism]. AB - OBJECTIVE: To explore the effect of sophora flavescens (SF) combined low molecular weight natural tumor suppressor(LMW-NTS) on normal human bone marrow CFU-GM and acute myelogenous leukemia cells and its mechanism. METHODS: CFU-GM colony culture, cell culture, Wrigh-Giemsa stain, 3H-TdR incorporation test, agarose gel electrophoresis of DNA fragment and RT-PCR were employed in this research. RESULTS: The combination of SF and LMW-NTS greatly inhibited the proliferation of acute meylogenous leukemia cells, induced their apoptosis, and down-regulated the c-Myc expression as compared with SF or LMW-NTS alone. CONCLUSION: The anti-leukemia effect of the combination of SF and LMW-NTS is superior to SF or LMW-NTS alone. PMID- 12575332 TI - [Experimental study on the apoptotic effect of arsenic trioxide on human osteosarcoma MG-63 cells]. AB - OBJECTIVE: To study the apoptotic effect of arsenic trioxide(As2 O3) on human ostrosarcoma MG-63 cells. METHODS: The MTT reduction test, transmission electron microscopy and agarose gel electrophoresis were applied to observe the apoptotic changes of human osteosarcoma line MG-63 induced by As2 O3. RESULTS: Typical apoptotic cellular changes were found after As2 O3 treatment. Transmission electron miroscopy showed that nucleolus disappeared and chromatin condensed under the nucleonic membrane. Agarose gel electrophorsis showed a "ladder" strand of DNA, a special phenomenon of cellular apoptosis. The above-mentioned changes were not found in the control group. CONCLUSION: As2 O3 can induce apoptosis of human osteosarcoma MG-63 cells. Further studies are to be carried out on its mechanism. PMID- 12575333 TI - [Pulmonary vascular remodeling in hypoxic hypertension in newborn pigs]. AB - OBJECTIVE: To determine the pulmonary vascular morphologic changes in small pulmonary arteries of normal development and hypoxic hypertension in newborn pigs. METHODS: Forty-two pigs aged from the birthday to 14 days were divided into the normal developmental group and hypoxic hypertensive group. Twenty of them were exposed to hypobaric hypoxia (50.8 kPa) for 3 or 11 days to establish a pulmonary hypertension model. With the immunohistochemistric method and tissue slices stained with gamma-actin antibody, small pulmonary artery morphologic changes were analyzed. RESULTS: In the normal newborn pigs, the percentage of media thickness decreased rapidly; and the ratio of the media area to the lumen area also decreased rapidly during the first 6 days after birth. Hypoxia weakened the decrease of the media thickness percentage and the ratio of the media area to the lumen area during the early period after birth. CONCLUSION: To adapt to the extrauterine life, newborn pigs rapidly remodel pulmonary blood vessels after birth. Hypoxia and hypoxic hypertensions induce abnormal pulmonary vascular remodeling causing pulmonary artery hypertension or making it serious. PMID- 12575334 TI - [Relationship between the expression of PTEN protein and phosphorylation of MAPK in hepatocellular carcinomas and their surrounding liver tissues]. AB - OBJECTIVE: To investigate the relationship between the expression of phosphatase and tensin homolog deleted on chromosome ten(PTEN) protein and phosphorylation of mitogen-activated protein kinase (MAPK) in hepatocellular carcinomas (HCC) and their surrounding liver tissues. METHODS: The expression of PTEN and the phosphorylation of MAPK were detected by the SP immunohistochemical technique in 75 cases of HCC and their pericarcinomatous tissues. RESULTS: The positive rate (62.7%) and the expression intensity of PTEN in HCC were lower and weaker than those in the pericarcinomatous liver tissues (89.3%)(P < 0.01), and the expression level of PTEN was significantly related to the differentiation degree of HCC. The poorer the differentiation of HCC, the weaker the expression of PTEN protein. The positive rate (85.3%) and the expression intensity of p42/44MAPK in HCC were higher and stronger than those in the pericarcinomatous liver tissues (34.7%)(P < 0.01), and the expression intensity of p42/44MAPK was not related to the differentiation degree of HCC. A significant negative relationship was observed between the expression intensity of PTEN protein and the phosphorylation level of MAPK in HCC and their surrounding liver tissues. CONCLUSION: The decrease or deletion of PTEN protein expression may not be able to effectively inhibit the over-activation of Ras/Raf/MAPK signaling pathway, and this will result in abnormal proliferation and malignant transformation of hepatocytes in hepatocellular carcinogenesis. PMID- 12575335 TI - [Experimental study on the viricidal effect of rough extracts of camptothecine fruit on herpes simplex virus-2 in vitro]. AB - OBJECTIVE: To study the viricidal effect of rough extracts of camptothecine fruit on herpes simplex virus-2 (HSV-2). METHODS: Rough extracts with different concentrations were mixed with HSV-2. The mixture was incubated at 37 degrees C for 4 h or at 4 degrees C overnight, adsorbed on monolayer cells of BGM for 1.5 h, and cultured at 37 degrees C for 7 d. RESULTS: The BGM of the test group did not reveal cytopathic effects (CPE), while the control group showed CPE under microscope observation. CONCLUSION: Camptothecine has a strong viricidal effect on HSV-2. PMID- 12575336 TI - [Effects of nao-yi-an granule on the intercellular expression of IL-6 in the experimental intracerebral hemorrhagic brain of rats]. AB - OBJECTIVE: To define the cellular localization of IL-6 and the effects of the traditional Chinese medicine complex nao-yi-an granule (NYA) on the intercellular expression of IL-6 in the experimental intracerebral hemorrhagic (ICH) brain of rats, so as to investigate the efficacy mechanisms of NYA on ICH. METHODS: ICH rat models were established by stereotaxically injecting 0.4 U type VII collagenase into the right globus pallidus (GP). The distribution and content of IL-6 protein expression together with the reaction of glial cells were investigated with the immunohistochemistry method. RESULTS: Microglial activation was detected from 6 h to 4 d after ICH in the hematoma region, and reactive astrocytes were found from 6 h to 7 d after ICH around the hematoma region; there was an up-regulation of IL-6 in the hematoma at 6 h after ICH; it reached its peak at 12 h, and disappeared on 7 d. CONCLUSION: The major sources of IL-6 positive protein were neurons and the activated microglia in the brain of ICH rats. Stabilizing the increasing expression of IL-6 may be one of the neuroprotective mechanisms of NYA. PMID- 12575337 TI - [Age related changes of serum sex hormone levels and tartrate-resistant acid phosphatase activity in healthy women]. AB - OBJECTIVE: To study the changes of serum sex hormone levels and tartrate resistant acid phosphatase (TRACP) activity in healthy women of different ages, and to provide parameters for basic and clinical research of osteoporosis. METHODS: Serum sex hormone (FSH, LH, E2, P) levels and TRACP activity were measured in 236 premenopausal healthy women aged 23-53 years and divided into 3 groups, and in 91 postmenopausal healthy women aged 48-71 years. RESULTS: Except serum FSH levels which were significantly higher in the third group than those of the other groups in premenopausal healthy women (P < 0.05), no significant difference in sex hormone levels and TRACP activity was found among the 3 groups; the postmenopausal group showed a significant increase in serum FSH, LH levels and TRACP activity and a significant decrease in serum E2, P levels, compared with the premenopausal groups (P < 0.01). There was a negative correlation between E2, P levels and TRACP activity in the postmenopausal women (r = -0.41 and -0.37, respectively, P < 0.01). CONCLUSION: Postmenopausal diminution of E2, P results in an increase of serum FSH and LH levels, and may bring about an increase of bone resorption, which results in the increase of TRACP activity. PMID- 12575338 TI - [Effect of losartan and amilodipine on the platelet activation and renal function in patients with mild and moderate hypertension]. AB - OBJECTIVE: To evaluate the effect of losartan and amilodipine on the platelet activation and renal function, and to study the relationship between the platelet activation and hypertensive renal damage in patients with mild and moderate hypertension. METHODS: Sixty patients with mild and moderate hypertension were divided into the losartan group and amilodipine group in a randomized and controlled method. Losartan or amilodipine was given for three months. The urinary albumin excretion rate (UAER), plasma alpha-granule membrane protein-140 (GMP-140), BUN, Bcr and creatinine clearance rate (CCR) were measured before and after the treatment. RESULTS: 1. The blood pressure, GMP-140 and UAER significantly decreased in the two groups after the treatment. 2. A positive correlation was found between the level of GMP-140 and that of UAER before or after the treatment in the patients with hypertension (r = 0.69, r = 0.48, P < 0.01), but no significant positive correlation was found between the decreased level of UAER and blood pressure after the treatment. CONCLUSION: The blood pressure can be well controlled, and the renal function is improved after the treatment with losartan or amilodipine, of which the inhibition to the patelet activation may be responsible for the improvement of the renal function in patients with mild and moderate hypertension. PMID- 12575339 TI - [Relationship between serum asymmetric dimethylarginine and blood pressure in patients with chronic renal failure]. AB - OBJECTIVE: To investigate the relationship between serum assymmetric dimethylarginine and blood pressure in patients with chronic renal failure. METHODS: We examined the levels of asymmetric dimethylarginine (ADMA) in the serum and urine of 29 hemodialysis patients using the HPLC method, and assessed the relationship between serum ADMA and blood pressure. RESULTS: The level of serum ADMA was higher, but the level of urinary ADMA was lower in the patients with chronic renal failure than those in the controls. The serum ADMA was positively related to blood pressure. CONCLUSION: The accumulation of ADMA may be related to hypertension in patients with chronic renal failure. PMID- 12575341 TI - [Two cases of type B Wolff-Parkinson-White syndrome in polytype]. PMID- 12575340 TI - [Cataract extraction through no-sitch small incision combined with intraocular lens implantation]. AB - OBJECTIVE: To investigate the method and clinical effect of cataract extraction through no-sitch small incision combined with intraocular lens (IOL) implantation. METHODS: Ninety-three cases of cataract were divided into two groups at random. Cataract extraction combined with IOL implantation through no sitch small incision was performed on one group, and cataract extraction combined with IOL implantation through conventional large incision was performed on the other group as controls. Follow-up was carried out regularly. RESULTS: The average uncorrected visual acuity of the small incision group at 1 week, 1 month, 3 months, 6 months and 1 year after the operation were significantly higher than those of the conventional large incision group. CONCLUSION: The small incision of cataract surgery results in a rapid and better rehabilitation of visual acuity. It needs no expensive instruments, costs less, and is suitable to be popularized. PMID- 12575342 TI - [Anterior approach in the surgical treatment of cervical fracture and spinal cord injury]. AB - OBJECTIVE: To explore value of the anterior approach in the surgical treatment of cervical fracture and spinal cord injury. METHODS: Thirty-one cases of cervical fracture and spinal cord injury were treated. All the patients were operated on in general anaesthesia by anterior decompression, reduction, auto-iliac graft and ORION cervical plate fixation. All the cases were evaluated according to Frankel criteria of the neural function preoperatively, and 3 months and 6 months after the operation. RESULTS: Twenty-eight cases were followed up for an average of 13.8 months (ranging from 3 to 18 months). All the fractures and dislocations were reduced satisfyingly. All the bone-grafts were fused within 3 months after the operation. There was no plate or screw complication in any of the cases. The Frankel grade improvement rates were 50%, and 82.1% in 28 cases at 3 months and 6 months postoperatively; the average Frankel grade was improved by a 0.64 and 1.14 grade respectively. CONCLUSION: For cervical fracture and spinal cord injury, an ideal reduction and immediate stability can be obtained through the anterior approach. Early surgery can significantly improve the spinal cord function and avoid missing the operation opportunity due to occurring complications. PMID- 12575343 TI - [Clinical characteristics of fatty livers of 61 patients with the insulin resistance syndrome]. AB - OBJECTIVE: To analyse the clinical characteristics of fatty livers of patients with the insulin resistance syndrome. METHODS: With echographic histogram, the acoustic intensity (AV) in six different areas of each fatty liver was measured in 61 patients with the insulin resistance syndrome. The relationship between the average value of AV and some clinical or biochemical parameters were analysed. RESULTS: There were significant differences in AV between the area near the portal venous and the area near the gall bladder, and between males and females as well. AV was positively related to the body mass ratio (BMI), waist circle (W) and hip circle (H); AV was positively related to the waist-hip ratio (WHR) or insulin sensitive index (ISI) in trend; and AV was negatively related to high density lipoprotein cholesterin (HDL-C). CONCLUSION: In patients with the insulin resistance syndrome, fatty density of livers is higher near the portal venous than near the gall bladder. The fatty liver is more serious in females than in males. BMI, W, H and serum HDL-C levels are useful parameters in evaluating the severity of fatty livers. PMID- 12575344 TI - [Twin test in the determination of alanine aminotransferase and aspartate aminotransferase in the serum]. AB - OBJECTIVE: To explore a method which can simultaneously determine alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum. METHODS: Using the self-prepared twin test reagent, ALT and AST in the serum were detected simultaneously. RESULTS: There was a good correlation between the twin test's method and the traditional method in the detection of ALT and AST (All r = 0.999). The precision of the method was good, and the coefficient variation (CV) was below 5%. Hemolysis, jaundice, and lipohemia serum were found to be interference-free. The time of the detection was two times of the traditional method. CONCLUSION: The twin test method can both exert the capability of the automatic analyzer and shorten the time of detection. It is a method worthy to be popularized. PMID- 12575345 TI - [Characteristics of the sleep apnea syndrome in the aged]. AB - OBJECTIVE: To study the characteristics of the sleep apnea syndrome in the aged. METHODS: Forty-two males who were suspicious of the sleep apnea syndrome (SAS) were detected with 7-hour polysomnography (PSG). RESULTS: The body mass index (BMI) and neck circumference showed no significant difference in the aged and middle aged groups. The apnea/hypopnea index (AHI) and microarousal index (MAI) were significantly lower, and the minimum oxygen saturation (SaO2) significantly higher in the aged than those in the middle aged (P < 0.05). Age was negatively correlated to AHI (r = -0.329, P < 0.05). The BMI and neck circumference were positively correlated to AHI (r = 0.326, P < 0.05; r = 0.306, P < 0.05, respectively). CONCLUSION: SAS patients are less severe in the aged excluding the affection of the BMI and neck circumference. PMID- 12575346 TI - [Clinical significance of the detection of the sL-selectin level in patients with acute leukemia]. AB - OBJECTIVE: To study the relationship between the episode and state of acute leukemia and the level of soluble L-selectin (sL-selectin) in the plasma and cerebrospinal fluid. METHODS: With a sandwich enzyme-linked immunosorbent assay (ELISA), the levels of sL-selectin in the plasma of 40 patients with acute leukemia and in the cerebrospinal fluid of 28 patients with acute lymphoblastic leukemia were measured, and compared with 20 controls. RESULTS: The levels of sL selectin were significantly higher in the patients with untreated and therapy resistant acute leukemia or leukemia relapse than those in the complete remission patients and the controls (P < 0.001). The levels of SL-selectin were related to the clinical course of acute leukemia. CONCLUSION: Monitoring the sL-selectin level may be useful for evaluating leukemia activity, in particular for the detection of leukemia relapse and meningeal infiltration. PMID- 12575347 TI - [Clinical analysis of arrthythmogenic right ventricular cardiomyopathy]. AB - OBJECTIVE: To investigate the clinical features of arrthythmogenic right ventricular cardiomyopathy (ARVC), and to evaluate the diagnosis of ARVC. METHODS: Twenty-three cases of ARVC underwent ECG, chest x-ray, Holter, transesophageal atrial pacing (TEAP) and intracardiac electrophysiological examination. RESULTS: A syncope attack occurred; the percentage of frequent ventricular premature beats was 95.7%, salvos of the right ventricular originated ventricular tachycardia (VT) was 87%, and the right bundle branch block (RBBB) demonstrated by ECG was 87%; UCG showed that the enlarged right ventricular diametric was (50.1 +/- 8.83) mm, and the right atrium diametric was (48 +/- 8.79) mm; the ratio of right ventricular diametric to left ventricular diametric in the end-diastolic period was 1.09. The right ventricular function [ejection fraction: (0.325 +/- 0.0902)] decreased, which was confirmed by echocardiogram. CONCLUSION: The diagnosis of ARVE can be established on the basis of the repeated syncope attack; RBBB, frequent ventricular premature beats, VT of LBBB patterms, an enlarged right heart and decreased right ventricular function can be found, the ratio of right ventricular dimaetric to left ventricular diametric increases in the end-diastolic period. PMID- 12575348 TI - [Congestive heart failure and paraoxonase]. AB - OBJECTIVE: To determine the changes of paraoxonase activity in patients with heart failure. METHODS: The serum paraoxonase (PON) activity of the heart failure group and controls was measured with phenylacetate as the substrate. RESULTS: The serum PON activity of the heart failure patients significantly decreased as compared with the controls; after the treatment, the PON activity of heart failure was largely elevated; and the PON activity of the patients with heart failure was negatively correlated with tumor necrosis factor-alpha. CONCLUSION: The PON activity of patients with congestive heart failure significantly decreases. PMID- 12575349 TI - [Expression of MMP1 and TIMP1 proteins in lung cancer and its biological significance]. AB - OBJECTIVE: To investigate the expression of MMP1 and TIMP1 proteins in lung cancer tissues and its biological significance. METHODS: MMP1 and TIMP1 proteins were detected with the immunohistochemical method of avidin-biotin complex in formalin-fixed and routinely paraffin-embeded specimens of 46 patients. RESULTS: There was no correlation in MMP1 protein expression between normal lung tissues (20%) and lung cancer (52.2%); The positive rate of TIMP1 protein expression was significantly lower in lung cancer (41.3%) than that in normal lung tissues (80%); There was a significant correlation among MMP1 and TIMP1 protein expression and differentiation, TNM stages, and lymph node metastases of lung cancer. There was a significant negative correlation between MMP1 and TIMP1 protein expressions in lung cancer. CONCLUSION: There is a correlation between MMP1 and TIMP1 protein expressions in lung cancer and the invasion and metastasis of lung cancer. The factors may be used as important biological markers in lung cancer. PMID- 12575350 TI - [Structure and regulative function of the 5' end flanking sequence of gene CCR5]. AB - OBJECTIVE: To construct the pCAT reporter genes containing the 5'-end flanking of CC chemokine receptor 5(CCR5) gene in different sequence lengths and identify the sequence, which regulates the gene expression of CCR5 by the CAT analysis system. METHODS: The target sequences were amplified by pyrobest DNA polymerase, and were inserted into the upstream of CAT gene located in the pCAT enhancer vector by the directional clone technique respectively; the regulative sequence was identified by analyzing the CAT activities of reconstructed plasmid in Hela cells. RESULTS: The region, containing 486 bp upstreaming from exon 1, stimulated the reporter gene activity which was about 3 times that of the pCAT enhancer vector in transfected cells. CONCLUSION: There is an up-regulative element of gene transcription in the region of -1(-)-486 bp in CCR5 gene upstream. PMID- 12575351 TI - [CT in the therapeutic evaluation of rare primary hepatic malignant tumors]. AB - OBJECTIVE: To discuss the significance of CT in the therapeutic evaluation of rare primary hepatic carcinoma. METHODS: The CT images and surgical data of 15 cases of rare primary hepatic malignant tumors were retrospectively analyzed. The correlative relationship between the CT images of common plus enhanced scan and the surgical operative difficulties was also analyzed. RESULTS: If CT images showed that huge lesions existed, severe lobles were involved, capsules were incomplete, the main branch of the portal venous system was involved, and the external structure of the liver was invaded before the operation, the operation might be more difficult, and complications and death might happen. CONCLUSION: CT images will directly indicate the difficulty of operation for rare primary hepatic malignant tumors before the operation, and an objective evaluation on the therapeutic method and operative effect can be made by CT examination. PMID- 12575352 TI - [Endoscopic sinus surgery in the treatment of fungal sinusitis]. AB - OBJECTIVE: To evaluate the curative effect of endoscopic sinus surgery on fungal sinusitis. METHODS: The clinical and follow-up data of 30 patients with fungal sinusitis who underwent endoscopic sinus surgery were analyzed retrospectively. RESULTS: Twenty-nine cases of non-invasive fungal sinusitis were cured by endoscopic sinus surgery; recurrence occurred in the 1 case of invasive fungal sinusitis two months after being treated by endoscopic sinus surgery and it was cured by lateral rhinotomy. CONCLUSION: The treatment of endoscopic sinus surgery is safe, effective, and less painful for patients with non-invasive fungal sinusitis. PMID- 12575353 TI - [Cerebrospinal fluid levels of beta-amyloid protein 1-42 in Alzhemier disease]. AB - OBJECTIVE: To study the role of beta-amyloid protein 1-42 (A beta 1-42) content in the cerebrospinal fluid (CSF) of Alzheimer disease (AD) patients. METHODS: A beta 1-42 levels were measured with the ELISA method in AD (n = 30), non-AD (NAD, n = 25) and non-dementia (ND, n = 21). RESULTS: The A beta 1-42 mean value for AD was (109.91 +/- 58.78) fmol.L-1. In ND, the A beta 1-42 mean value was (242.40 +/ 142.58) fmol.L-1. The mean value for AD was significantly lower than that of ND. In NAD, the A beta 1-42 mean value was (231.70 +/- 143.94) fmol.L-1, and it was not significantly different from the mean value for ND. The A beta 1-42 level was positively correlated with the severity of AD symptoms, but not with the duration. A beta 1-42 levels in CSF of AD were significantly lower than that of ND, and they decreased as the severity of disease increased. CONCLUSION: Cerebrospinal fluid beta-amyloid 1-42 analyses may be of value in the clinical diagnosis of Alzheimer's disease, especially in the earlier course of the disease, when drug therapy may have the greatest effect but clinical diagnosis is particularly difficult. PMID- 12575354 TI - [Effect of 99Tc-MDP on cytokine production by peripheral blood mononuclear cells of patients with rheumatoid arthritis]. AB - OBJECTIVE: To study the immune mechanism of 99Tc-MDP in the treatment of patients with rheumatoid arthritis (RA). METHODS: In vitro, 99Tc-MDP was added to the cultures of peripheral blood mononuclear cells (PBMC) derived from healthy subjects and RA patients. The levels of IL-1 and sIL-2R from PBMC affected by 99Tc-MDP were determined with enzyme linked immunosorbert assays (ELISA). RESULTS: The IL-1 production by PBMC of RA, either in the medium alone (spontaneously) or in the presence of lipopolysacchride (LPS) stimulation, was suppressed by 99Tc-MDP; and the sIL-2R by PBMC of RA, either in the medium alone (spontaneously) or in the presence of phytohemagglutinin (PHA) induction, was inhibited by 99Tc-MDP. CONCLUSION: 99Tc-MDP acts on PBMC and a possible immune activity by 99Tc-MDP is related to the suppression of IL-1 and sIL-2R in RA patients. PMID- 12575355 TI - [A case of non-Hodgkin's lymphomas metastasize to the cardiac right atrium]. PMID- 12575356 TI - [Application of SPECT bone scan for multiple myeloma]. AB - OBJECTIVE: To study the diagnostic value of SPECT for multiple myeloma (MM). METHODS: Single photon emission computed tomography (SPECT) bone scan, marrow puncturing, x-ray and other related examinations were performed in 46 patients with MM. We analysed the diagnostic value of SPECT bone scan combined with marrow puncturing and other examinations for MM. Some patients were examined with SPECT bone scan in order to estimate the efficacy of MM. RESULTS: SPECT bone scan images of 44 patients were abnormal in the 46 patients with MM. The masculine rate was 95.6%. In most of the patients, there was multiple abnormal osseous metabolism in the whole body, especially in the spine, double ribs, and pelvis. We observed the efficacy of 2 patients with SPECT bone scan. After the therapy, SPECT bone scan was performed again in 1 patient, and the image showed that the range of abnormal osseous metabolism was reduced; SPECT bone scan was performed again in the other patient, and the abnormal range was found to have increased obviously. CONCLUSION: SPECT bone scan is one of the important assistant examination methods for MM. Its sensitivity is high. The nuclear medical diagnoses should be combined with clinical documents. SPECT bone scan might be available for monitoring the efficacy of MM. PMID- 12575357 TI - [Implantable cardioventer defibrillator in the treatment of malignant ventricular arrhythmias and its short-term follow-up]. PMID- 12575358 TI - [A case of the primary malignant lymphoma of the liver]. PMID- 12575359 TI - [Expression of caspase 3 in gastric carcinoma and atypical proliferation and its association with apoptosis]. PMID- 12575360 TI - [Changes of cardiac inside diameter and ejection fraction in patients with heart failure after the treatment with nitroprusside]. PMID- 12575361 TI - [A case of left coronary artery fistula into the pulmonary artery with massive hemoptysis]. PMID- 12575362 TI - [Bronchial artery angiography and embolization in the treatment of bronchiectasis and hemoptysis]. PMID- 12575363 TI - [Curative effect of the sheng-mei injection on coronary disease and angina pectoris]. PMID- 12575365 TI - [Cerebral infarction following to total hip replacement]. PMID- 12575364 TI - [Complication and its treatment of post-cranioplasty with silastic]. PMID- 12575366 TI - [A case of lung cancer updating 99mTc-MDP]. PMID- 12575367 TI - [Relation of the four famous doctors in Beijing and integrated traditional Chinese and Western medicine]. PMID- 12575368 TI - [Multiple center clinical study on endocrinologic drugs must be conducted according to the national norm of clinical trial of drugs]. PMID- 12575369 TI - [Looking back on some matters of integrated traditional Chinese and Western medicine]. PMID- 12575370 TI - [Clinical study on effect of tongyu no. I in improving prethrombotic state of senile diabetes mellitus]. AB - OBJECTIVE: To explore the therapeutic mechanism of Tongyu No. I (TY-1) in improving prethrombotic state of senile diabetes mellitus. METHODS: Sixty-two patients of diabetes mellitus with prethrombotic state and blood-stasis Syndrome were selected and divided randomly into two groups, 32 in the treated group and 30 in the control group. The two groups were treated by conventional therapy, combined with additional TY-1 (treated group) and aspirin (control group) respectively for 2 months. And the clinical effect as well as relevant criteria in both groups before and after treatment were observed. RESULTS: In the treated group, treatment showed markedly effective in 14 cases, effective in 16 and ineffective in 2, the total effective rate being 93.8%. In the control group, the corresponding values were 6, 14, 10 and 66.7% respectively. Comparison of the total effective rate in the two groups showed significant difference (P < 0.01). After treatment, the treated group showed lowering in plasma TXB2 similar to that in the control group, while the decrease of GMP-140 and fasting blood glucose levels was significant as compared with before treatment (P < 0.01) and the difference was significant between the two groups (P < 0.05) after treatment. CONCLUSION: TY-1 could improve the prethrombotic state and blood-stasis Syndrome in patients of diabetes mellitus, the main therapeutic mechanism might be correlated to the lowering of GMP-140 content. PMID- 12575371 TI - [Clinical and experimental study on anti-liver fibrosis effect of xuelong granule]. AB - OBJECTIVE: To study the clinical effect and mechanism of Xuelong Granule (XLG) in antiliver fibrosis. METHODS: Ninety-eight patients with liver cirrhosis were divided into 2 groups randomly. The 58 Patients in the treated group were treated by XLG and the 40 patients in the control group were treated by Shenchai Granule for 3 months. Levels of serum hyaluronic acid (HA) and laminin (LN), and pathological changes of liver tissues were observed before and after treatment. In experimental study on model rats, the liver tissue content of hydroxyproline and pathological changes under light and electron microscope were observed. RESULTS: The total effective rate in the treated group was 72.4%, that in the control group was 40.0%, there was significant difference between the two groups (P < 0.01). The level of serum HA and LN in the treated group declined markedly after treatment, and pathological examination of 8 cases showed the fibrous tissues reduced obviously. Experimental study showed marked reduction of hydroxyproline content and significant lowering of fibrous tissue proliferation, both under light and electron microscope, in liver of model rats after XLG treatment. CONCLUSION: XLG has a definite effect in anti-liver fibrosis. PMID- 12575372 TI - [Clinical study on treatment of chronic bronchitis by tracheitis plaster]. AB - OBJECTIVE: To study the clinical effect and mechanism of Tracheitis Plaster (TP) in treating chronic bronchitis. METHODS: TP is consisted of ephedra, almond, pinellia tuber, earthworm and white mustard seed. Patients were randomly divided into two groups, 59 patients in the treated group were treated with TP sticking on acupoints Dingchuan, Dashu, Fengmen, Feishu and Xinshu at back along both sides of thoracic vertebrae 1-6 and the 25 patients in the control group were treated with intramuscular injection of Siqikang. The times of treatment for both groups were 20. Clinical symptoms, X-ray chest film, level of immunoglobulin and T-lymphocyte subsets were recorded before and after treatment, and follow-up were carried out 0.5-1 year later. RESULTS: The clinical total effective rate was 93.2% and the X-ray improvement rate was 40.7% in the treated group, while in the control group, 80.0% and 20.0% respectively. Half and 1 year follow-up studies showed the total effective rate in the treated group was 91.5% and 89.8% respectively, which was significantly higher than that in the control group (80.0% and 76.0%) respectively (P < 0.05). The improvement in levels of IgG and CD8 in the treated group was also superior to those in the control group (P < 0.05). CONCLUSION: TP is a highly effective transcutaneous absorbent with promising long-term effect, it could regulate the immune function. PMID- 12575373 TI - [Clinical observation on treatment of asthma in moderate degree with fluticasone inhalation combined with jinacon]. AB - OBJECTIVE: To observe the efficacy of Fluticasone inhalation combined with Jinacon (JNC) orally taken in treating patients of chronic asthma (CA) of moderate degree. METHODS: Seventy-two CA patients were divided into three groups randomly and treated with Fluticasone inhalation combined with JNC, Fluticasone inhalation alone and JNC alone respectively for 4 weeks. The changes of clinical symptoms, airway reactivity and pulmonary function after treatment were observed. RESULTS: The clinical symptoms, pulmonary function and airway hyper-reactivity were improved in all the three groups after treatment (P < 0.05 or P < 0.01), but the improvement was more significant in the group treated with the combined therapy (P < 0.05 or P < 0.01). CONCLUSION: Fluticasone inhalation combined with JNC orally taken is an effective therapy in treating patients of chronic asthma of moderate degree. PMID- 12575374 TI - [Effect of new zhengtian pill on expression of whole blood platelet membrane adhesion molecules in patients of migraine]. AB - OBJECTIVE: To investigate the effect of New Zhengtian Pill (NZTP) on expression of whole blood platelet membrane adhesion molecules (PMAM) in patients of migraine. METHODS: Sixty-eight patients were divided into two groups, the 35 patients in the treated group treated by NZTP orally and the 33 patients in the control group treated by Fuguiqin Capsule with a therapeutic course of 30 days for both groups. Changes of PMAM GP II b/III a(CD41) and P-selectin (CD62P) were observed by flow-cytometry and compared with those in healthy persons. RESULTS: The markedly effective rate and total effective rate in the treated group was higher than those in the control group respectively (P < 0.05 and P < 0.01). The PMAM expression was also higher in patients, both at onset stage and intermittent stage, than in healthy persons (P < 0.01), NZTP treatment could reduce their increased expression significantly (P < 0.01). CONCLUSION: NZTP could reduce the PMAM expression and inhibit the activation of platelet. PMID- 12575375 TI - [Effect of qidan tongmai tablet on glucose and lipid metabolism in patients with diabetes mellitus type 2]. AB - OBJECTIVE: To study the effect of Qidan Tongmai tablet (QDTMT) on glucose and lipid metabolism in patients with diabetes mellitus (DM) type 2. METHODS: Patients of DM, with or without hyperlipidemia complication (HLC) were divided into 4 groups, Group A (33 cases without HLC) and B (33 cases with HLC) were treated by QDTMT, Group C (31 cases without HLC) and D (31 cases with HLC) were not treated by QDTMT. The treatment was carried out on the previous basic hypoglycemic treatment with a therapeutic course of 2 months. RESULTS: The levels of fasting glucose, 2 hrs postprandial blood glucose and glycosylated hemoglobin, as well as the levels of total cholesterol and triglyceride lowered, and the high density lipoprotein increased in Group A and B after treatment, as compared with those before treatment, the difference was significant (P < 0.01). While in Group C and D, the above-mentioned indexes were not changed significantly (P > 0.05). No apparent side-effect was found in the QDTMT treatment period. CONCLUSION: Besides regulating abnormal blood lipid, QDTMT has also a hypoglycemic effect in certain degree in patients with DM type 2. PMID- 12575376 TI - [Clinical observation on effect of xuejie jiawei decoction on vaginal hemorrhagic amount and duration in patients after drug-abortion]. AB - OBJECTIVE: To observe the effect of Xuejie Jiawei Decoction (XJJWD) in reducing vaginal hemorrhagic amount, shortening hemorrhagic duration and lowering the curettage rate of incomplete abortion in patients after drug-abortion. METHODS: Group A was treated with XJJWD No. I from the 2nd day after abortion, one dose per day for 7 days. Group B was treated with XJJWD No. II from the 11th day after abortion, one dose per day for 5 days. The therapeutic effect of the two groups was compared with that of the untreated control group. RESULTS: In the Group A, the total score on vaginal hemorrhagic amount was markedly less than that in the control group (P < 0.01), and the vaginal hemorrhagic duration in patients with menostasis less than 45 days was shorter than that in the control group (P < 0.01). In the Group B, the total score on vaginal hemorrhagic amount in patients with menostasis less than 45 days was lower than that in the control group (P < 0.05). CONCLUSION: XJJWD could reduce the vaginal hemorrhagic amount and duration in patients after drug-abortion. It should be applied early after abortion, the less the duration of menostasis, the more effective the treatment. PMID- 12575377 TI - [Immune modulatory and therapeutical effect of shenqi tablet accessory therapy in treating recurrent genital herpes]. AB - OBJECTIVE: To observe the immune modulatory and therapeutical effect of Shenqi tablet (SQT) in treating recurrent genital herpes. METHODS: Sixty-three patients were randomly divided into two groups, the SQT group (34 cases) and the control group (29 cases). The immunologic function of patients was determined before and after treatment and the recurrence rate of two groups was compared. RESULTS: In the SQT group after treatment, CD3, CD4 and CD4/CD8 ratio raised, CD8 percent lowered, serum interleukin 2 (SIL-2) and RBC-C3b raised and serum SIL-2 receptor lowered (all P < 0.05), while in the control group, the above-mentioned parameters were not changed significantly. Follow-up conducted 6 months after treatment showed that the recurrence rate in the SQT group was 26.5% (9 cases), which was lower than that in the control group (72.4%, 21 cases) significantly (P < 0.01). CONCLUSION: SQT has immune modulatory effect in patients with recurrent genital herpes, it could reduce the recurrence rate. PMID- 12575378 TI - [Regulatory effect of tiaoxin recipe drug serum on animal's Alzheimer disease related tau protein phosphorylation]. AB - OBJECTIVE: To study the regulatory effect and possible mechanism of Tiaoxin Recipe (TXR) on animal's Alzheimer disease related tau protein phosphorylation. METHODS: NG108 cell model was treated with Okadaic acid and related parameters were determined using MTT staining, immunoblot, coimmunoprecipitation assay, etc. RESULTS: Shown by MTT staining, NG108 cell activity decreased significantly after treated with Okadaic acid for 12 hrs, which could be ameliorated by TXR rat serum. Revealed by immunoblot method, the Okadaic acid induced elevation of phosphorylated tau protein could partly be reversed after co-treated with TXR rat serum. TXR extract could inhibit the binding of tau protein with presenilin-1, which may regulate the tau protein phosphorylation, and could be observed by coimmunoprocipitation. CONCLUSION: TXR could inhibit tau protein hyperphosphorylation, which might partially be due to the TXR caused binding of presenilin-1 with tau protein. PMID- 12575379 TI - [Experimental pharmacodynamic study on the anti-convulsion effect of shenpu decoction]. AB - OBJECTIVE: To observe the anti-convulsion effect of Shenpu Decoction (SPD). METHODS: Experiments were conducted in three acute convulsion models (cardiazol seizure, strychnine convulsion and maximal electrical shock). Models were divided into the control group (A, treated by normal saline), the high, middle and low dose SPD groups (B, C and D, treated with SPD 9 g/kg, 6 g/kg and 4 g/kg respectively), and the nitrazepam treated group (E). The anti-convulsion effect of SPD was evaluated by 50% convulsion dose (CD50) detected in each group. RESULTS: In cardiazol convulsion model, the CD50 detected in group A-E were 63.3 +/- 3.4 mg/kg, 116.2 +/- 3.4 mg/kg, 105.6 +/- 3.7 mg/kg, 74.0 +/- 3.7 mg/kg and 197.2 +/- 3.3 mg/kg respectively, while they were 0.71 +/- 0.04 mg/kg, 1.21 +/- 0.04 mg/kg, 1.19 +/- 0.04 mg/kg, 0.94 +/- 0.04 mg/kg and 1.16 +/- 0.04 mg/kg respectively in the strychnine convulsion model, and 67.1 +/- 2.6 V, > 140 V, 109.4 +/- 3.5 V, 84.4 +/- 3.1 V and 128.4 +/- 3.9 V in the maximal electrical shock model respectively. CONCLUSION: SPD has a good anticonvulsion effect. PMID- 12575380 TI - [Study on effect of qingkailing injection and its active principle in inducing cell apoptosis in human acute promyelocytic leukemia]. AB - OBJECTIVE: To investigate the mechanism of Qingkailing (QKL) Injection in treating acute promyelocytic leukemia. METHODS: Using MTT technique, cell morphologic method, DNA gel electrophoresis and flow-cytometry to study the human acute promyelocytic leukemia (HL-60) cell apoptosis induced by QKL and its active principle. RESULTS: QKL and its active principle, Baicalin and hyodeoxycholic acid, showed strong cytotoxicity in inhibiting HL-60 cell, the Bezoar cholic acid showed a weaker effect. Apoptosis could be induced after being treated for 6 hrs by the former three principles, showing a typical apoptosis peak under flow cytometry, but could not be induced by the latter one. CONCLUSION: QKL could induce leukemia cell apoptosis in vitro, which may be one of the mechanisms of QKLI in curing acute promyelocytic leukemia. PMID- 12575381 TI - [Study on effect of jinshui liujun jian oral liquid on serum superoxide dismutase activity and malonyldialdehyde content in mice with chronic bronchitis]. AB - OBJECTIVE: To study the effect of Jinshui Liujun Jian Oral Liquid (JLJOL) on serum superoxide dismutase (SOD) activity and malonyldialdehyde (MDA) content in mice with chronic bronchitis. METHODS: JLJOL was given to the chronic bronchitis mice model (induced by smoking) through gastrogavage, and then SOD activity and MDA content were tested. RESULTS: SOD activity in model mice after JLJOL treatment was 0.67 +/- 0.15 NU/L, which was significantly higher than that in the untreated model (0.39 +/- 0.13 NU/L, P < 0.01). But the MDA content in treated mice was significantly lower than that in untreated one (9.26 +/- 2.90 nmol/L vs 16.07 +/- 5.62 nmol/L, P < 0.01). CONCLUSION: JLJOL could scavenge the injury of free radical on organism. PMID- 12575382 TI - [Effect of conventionally prepared rabbit serum on human umbilical vein endothelial cell-304]. AB - OBJECTIVE: To study the effect of conventionally prepared rabbit serum, usually used in the field of serum pharmacology of TCM, in various concentration on cultured system of human umbilical vein endothelial cell (VEC)-304 cell line. METHODS: Prepared conventionally from healthy rabbit, the effect of uncontaminated and nonhemolytic serum of six concentrations (5%, 10%, 20%, 40%, 60% and 80%) on VEC-304 cells was observed using parameters such as cell morphologic examination, trypan blue exclusion test, methyl thiazolyl tetrazolium (MTT) test, lactate dehydrogenase (LDH) and angiotensin-converting release test. RESULTS: Serum of 5% and 10% concentration showed no effect in all the above mentioned tests. Serum of 20% concentration also showed no effect in the tests except for LDH releasing (P < 0.01). But when the concentration of serum reached up to 40% or above, distinct cytotoxicity and serious injury appeared in a dose dependent manner. CONCLUSION: High concentration of conventionally prepared rabbit serum could injure the incubated cell. Therefore, in order to lower the background effect of serum as far as possible in TCM serum pharmacological study, the cytotoxicity of rabbit serum should be tested first of all. PMID- 12575383 TI - [Experimental study on effect of congsheng capsule on expression of brain derived neurotrophic factor and tyrosinkinase mRNA in forebrain of rats after cortical devascularization]. AB - OBJECTIVE: To study the effect and mechanism of Congsheng Capsule (CSC) on brain derived neurotrophic factor (BDNF) and tyrosinkinase (trkB) mRNA in forebrain of rats after cortical devascularization. METHODS: The rat devascularization model was established, and expression of BDNF mRNA and trkB mRNA were determined by hybridization in situ method. RESULTS: Expression of BDNF mRNA and trkB mRNA reduced obviously in the cerebral cortex and hippocampus of devascularization model, especially in hippocampal CA1 subregion. There was trkB mRNA but no BDNF mRNA expression in the macrocyte basal nuclei (MBN). CSC significantly increased the decreased BDNF and trkB mRNA in cortex, hippocampus and MBN compared with placebo group. CONCLUSION: CSC could salvage the degenerating neurons and maintain their survival after cortical devascularization by increasing the expression of BDNF and trkB mRNA and improving the synthesis of BDNF mRNA and trkB protein. PMID- 12575384 TI - [Progress of TCM study on insulin resistance]. PMID- 12575385 TI - [Study on Chinese herbal medicine induced leukemic cell apoptosis]. PMID- 12575386 TI - [Chinese Journal of Integrated Traditional and Western Medicine and I]. PMID- 12575387 TI - [The cause of integrated traditional Chinese and Western medicine and I]. PMID- 12575389 TI - [Effect of sodium ferulate on leucocyte spontaneous activation and surface adhesion molecule expression in arteriosclerosis obliterans patients]. AB - OBJECTIVE: To explore the effect of sodium ferulate (SF) on leucocyte spontaneous activation rate (LSAR) and the surface expression of adhesion molecule (SEAM) in patients with arteriosclerosis obliterans (ASO). METHODS: Forty-two patients with ASO were treated by SF, LSAR and SEAM in them were determined qualitatively before and after SF treatment, as well as in 36 healthy subjects for control. RESULTS: Therapeutic effect of SF in the treated 42 patients was short-term cured in 17 cases (40.5%), markedly effective in 21 (50%), effective in 2 (4.8%), ineffective in 2(4.8%), and the total effective rate being 95.2%. LSAR and SEAM in ASO patients before treatment was 14.84 +/- 7.54% and 47.94 +/- 13.48% respectively, which was obviously higher than 4.96 +/- 2.04% and 24.12 +/- 9.35% in healthy subjects (P < 0.01) respectively, but markedly reduced to 8.19 +/- 4.09% and 28.63 +/- 7.91% after SF treatment (P < 0.01). CONCLUSION: SF could not only obviously lower both LSAR and SEAM but also exert a good therapeutic effect on patients with ASO. PMID- 12575390 TI - [Clinical study on effect of qihuang oral liquid to enteric flora disturbance and serum endotoxin level in patients of liver cirrhosis]. AB - OBJECTIVE: To study the effect of Qihuang oral liquid (QHOL) in treating enteric flora disturbance and serum endotoxin level of liver cirrhosis patients. METHODS: Seventy patients suffering from liver cirrhosis were randomized into the control group and the QHOL treated group. The symptomatic changes, quantitative determination of anaerobic and aerobic flora in feces as well as serum endotoxin level were observed before and after treatment. RESULTS: In comparison with the control group, the treated group revealed a significantly better effects (P < 0.05) in the following parameters: (1) reduction in aerobic and increase in anaerobic flora, thus to improve the ratio of enteric flora; (2) improvement in clinical symptoms; (3) lowering in serum endotoxin level. CONCLUSION: QHOL might alter the ratio of enteric flora by increasing anaerobics, a significant lowering of enterogenic endotoxin production and absorption, hence obviously reduced serum endotoxin level was induced which might correlate to the improvement in symptoms and liver damage. PMID- 12575391 TI - [Clinical study on effect of hewei capsule on promoting gastric dynamics]. AB - OBJECTIVE: To investigate the effect of Hewei capsule (HWC) on gastric dynamics. METHODS: Sixty-two patients with dysmotility-like functional dyspepsia (DFD) were included in the study. The principal symptoms, electrogastrogram (EGG), plasma motilin (MTL), serum gastrin (Gas) and nitric oxide (NO) of the patients were the chief parameters observed before and after treatment. RESULTS: There were arrhythmia of EGG, deficiency of MTL, Gas and NO in the patients with DFD, and these abnormal changes could be improved significantly by treating with HWC. CONCLUSION: The abnormal gastric dynamics of the patients with DFD could be improved by HWC through multiple pathways. PMID- 12575392 TI - [Clinical study on effect of recombinant roasted licorice decoction combined with low-dose glucocorticoids in treating idiopathic thrombocytopenic purpura]. AB - OBJECTIVE: To explore the clinical effect of recombinant roasted licorice decoction (RRLD) combined with low-dose glucocorticoids (GC) in treating idiopathic thrombocytopenic purpura (ITP). METHODS: Forty-three patients were divided semi-randomly into the treated group (n = 21) and the control group (n = 22), the former was treated with RRLD plus low-dose GC, and the latter was treated mainly with normal dosage of GC. RESULTS: The total effective rate of the treated group was better than that of the control group, the difference between these two groups was significant (P < 0.05). After treatment the platelet count in the treated group arose from (36.29 +/- 9.70) x 10(9)/L to (95.14 +/- 18.26) x 10(9)/L, while in the control group from (37.31 +/- 9.29) x 10(9)/L to (77.73 +/- 21.96) x 10(9)/L, the RRLD was superior to nat of GC in raising platelet count, the difference between them was also significant (P < 0.01). The treated group was lower obviously than the control group in adverse reaction (P < 0.01). CONCLUSION: The efficacy of RRLD plus low-dose GC is reliable in treating ITP, with less adverse reaction. PMID- 12575393 TI - [Study on using colloidal gold immuno-dot assay to detect special antibody of hemorrhagic fever renal syndrome]. AB - OBJECTIVE: To seek for a simple, sensitive and rapid assay to detect specific antibody of patients suffering from hemorrhagic fever renal syndrome (HFRS). METHODS: Serum anti-HFRS IgM (sIgM) and IgG (sIgG) antibody were detected in 186 patients with HFRS using colloidal gold immuno-dot assay (CGIDA), and compared with those detected with enzyme linked immunosorbant assay (ELISA) and indirect fluorescent antibody test (IFAT). Kuhuang Shenmai injection (KHSM) combined Astragalus decoction (AD) treatment was applied to 101 HFRS patients in a treated group (n = 50), and the effect of therapy was compared with that in a control group (n = 51) treated with ribarvirin and ganlixin injection. RESULTS: CGIDA showed positive sIgM antibody in 132 cases (70.9%) and positive sIgG antibody in 163 (87.1%) of the 186 HFRS patients. As compared the two groups in crossing stage, the rate of crossing shock stage in the treated group was higher than that in the control (P < 0.05). As for rate of crossing oliguria stage and from febrile stage directly developed into diuretic stage, the difference between the two groups was insignificant (P < 0.05). CONCLUSION: CGIDA shows the merits of quick, simple and convenient in detecting sIgM and sIgG antibody in HFRS patients. The effect of KHSM combined AD treatment was not significantly different from that of ribarvirin and ganlixin injection. PMID- 12575394 TI - [Perspective study on effect of gandou tablet I on biliary trace elements in treating hepatolenticular degeneration]. AB - OBJECTIVE: To observe the effect of gandou tablet I (GDI), a Chinese herbal recipe, on the content of biliary trace elements in hepatolenticular degeneration(HLD) patients. METHODS: Before and after 4 weeks treatment by oral taken GDI, bile of 32 cases of HLD were collected by duodenal drainage for determining biliary trace elements, including copper, zinc, iron and calcium, by spectrophotometer of atom absorption spectrum, and compared with those of 30 cases of non-HLD patients as the control. RESULTS: The contents of copper, iron and copper/zinc ratio were obviously lower, while the content of calcium greatly higher (P < 0.01), in the HLD group before GDI treatment than those in the control (P < 0.01 or P < 0.05), but the content of zinc was not significantly different between the two groups. The output of biliary copper after GDI treatment was significantly increased compared with that before treatment (P < 0.05), but the contents of zinc, iron, calcium and copper/zinc ratio were not markedly changed. CONCLUSION: The impediment to output of biliary copper was one of the mechanisms for copper storage in body of HLD patients. GDI could promote excretion of biliary copper in the HLD patients. PMID- 12575395 TI - [Study on effect of spleen invigorating, qi benefiting and dampness removing Chinese drugs and Western medicine on serum epidermal growth factor in chronic atrophic gastritis patients]. AB - OBJECTIVE: To investigate the effect of various combinations of Spleen invigorating and Dampness removing recipes and western medicine on level of serum epidermal growth factor (sEGF) in patients of chronic atrophic gastritis (CAG). METHODS: By means of Syndrome Differentiation three groups were divided as (1) Spleen invigorating I group (SI-I); (2) Spleen invigorating II group (SI-II); (3) Dampness removing group (DR); (4) western medicine group (WM); (5) healthy volunteers served as normal control group. The changes of symptoms and sEGF level were determined and compared between groups as well as before and after treatment. RESULTS: The sEGF in various groups of CAG were higher than that in the normal control (P < 0.01). In patients of SI-I and II groups, symptoms and pathological manifestations were improved significantly (P < 0.01), and sEGF dropped markedly after treatment (P < 0.01 and P < 0.05). The level of sEGF in DR raised after treatment (P < 0.05), but the level of sEGF in WM group lowered insignificantly. CONCLUSION: Inflammation of gastric mucosa could cause responsive elevation of sEGF in CAG patients. After treated with Spleen invigorating and Dampness removing Chinese drugs, the symptoms of CAG improved, simultaneously with the restoration of sEGF. sEGF could be taken as a sensitive index of the prognosis of CAG. PMID- 12575396 TI - [Clinical study on therapeutic effects of treatment according to syndrome differentiation of traditional Chinese medicine combined with captopril on severe viral myocarditis complicated heart failure]. AB - OBJECTIVE: To observe the therapeutic effect and mechanism of the treatment according to Syndrome Differentiation of TCM combined with captopril (CAP) on severe viral myocarditis (SVM) complicated heart failure (CHF). METHODS: One hundred and nine patients of SVM complicated CHF were randomly divided into the treated group (n = 72) and the control group (n = 37), the former was treated with TCM combined with CAP, while the latter was treated with dexamethasone and interferon. The TCM prescriptions were made depending on types of diseases by Syndrome Differentiation, i.e. Heart-Spleen deficiency type, Qi-Yin deficiency type and Spleen-Kidney Yang deficiency type. The efficacy of treatment was evaluated by the criteria including NYHA classification, myocardial enzymology, electrocardiogram, cardiac function and motorial toleration measured before and after treatment. RESULTS: The therapeutic effect of the treated group according to NYHA classification was obviously better than that of the control group; the creatine phosphokinase isoenzyme (CPK-MB), aspartate transaminase (AST), lactate dehydrogenase (LDH) content lowered in both groups, but more significantly lowered in the treated group than in the control group (P < 0.05, P < 0.01); the improvement of S-T segment of ECG in the treated group was better than that in the control (P < 0.01); also some parameters of heart function and motorial toleration were bettered in the treated group more significantly (P < 0.01). CONCLUSION: TCM treatment according to Syndrome Differentiation combined with CAP in treating SVM complicated CHF could elevate the clinical efficacy. PMID- 12575397 TI - [Clinical study on analgesic mechanism of bitongxiao in treating neck pain due to cervical spondylitis]. AB - OBJECTIVE: To study the analgesic mechanism of Bitongxiao (BTX, a Chinese herbal decoction) in treating neck pain due to cervical spondylitis (CS). METHODS: BTX and Jingfukang (JFK) granule were used to treat the treated and the control group separately in 102 CS cases. Plasma leucine enkephalin (LEK), substance P (SP) and prostaglandin E2 (PGE2) before and after treatment were assayed by radioimmunoassay (RIA). RESULTS: Twelve patients in the treated group were clinically cured, 36 markedly effectively, 3 effectively and 1 ineffectively, while in the control group they were 5, 33, 14 and 0 cases respectively. The cured and marked effective rate in the treated group was 92.3%, while that in the control group 72.0%. After treatment the pain score (VAS) of both groups were significantly reduced, but the reduction was higher in the treated group than that in the control. Before treatment, plasma LEK in all patients was abnormally low, but after treatment, it raised obviously more in the treated group than that in the control (P < 0.05). Plasma SP of both groups were obviously elevated, and PGE2 revealed unaltered after treatment. CONCLUSION: BTX has strong, fast-acting and long-lasting analgesic effect with no side-effects, it could elevate the lowered plasma LEK level in cervical spondylitis patients and increase plasma SP level. The analgesic mechanism of BTX might be associated with its effect on LEK, SP or other endorphins, directly acting on the pain modulation system. PMID- 12575398 TI - [Effect of tiangui recipe on hypothalamic proopiomelanocortin mRNA levels in androgen sterilized rats]. AB - OBJECTIVE: To explore the effect of Tiangui recipe (TGR) in androgen sterilized rats (ASR). METHODS: New-born female SD rats were randomly divided into 3 groups. ASR model was established by means of testosterone injection subcutaneously at the age of 9 days, ASR were administered with TGR for 14 days. Around the age of 106 days (proestrus day), all rats were sacrificed, serum total testosterone (TT) and free testosterone (FT) were measured by radioimmunoassay (RIA), and levels of hypothalamic proopiomelanocortin (POMC) mRNA expression in hypothalamus were determined. RESULTS: None of the ASR ovulated and all rats in control group ovulated. The rate of ovulation in TGR group was 58%, the mean-serum concentration of TT and FT in ASR were significantly higher than those in the control group and TGR group (P < 0.01). The hypothalamic POMC mRNA level in ASR model group was significantly higher than that in the control group (P < 0.01), and became normal after TGR treatment. CONCLUSION: The elevated serum androgen levels in ASR could challenge hypothalamic POMC mRNA overexpression via androgen receptor (AR). TGR might reduce serum androgen and lower the expression of POMC mRNA. PMID- 12575399 TI - [Effect of bushen huoxue decoction on in vitro fertilization and early embryonic development in mice]. AB - OBJECTIVE: To explore the effect of Bushen Huoxue decoction (BSHX) on female reproduction and elucidate its therapeutic mechanism to infertility. METHODS: The BSHX medicated serum of rabbit, as a supplement, was co-cultured with the sperm and ovum of non-copulated mice, and the 2-cell embryos of copulated female mice separately, to observe the changes of in vitro fertilization rate (IVF) and early embryogenesis rate. RESULTS: By co-cultured with BSHX medicated serum, IVF rate was increased obviously (P < 0.01), and the follow-up early embryogenesis rate at various period was promoted, particularly that of the 4-cell and 8-cell embryos (P < 0.05). No influence on developmental rate of in vitro 4-cell embryos obtained from the in vivo 2-cell embryos. But the development of the 8-cell embryos, morula, blastula and hatching were promoted significantly. CONCLUSION: BSHX could raise the fertilization rate and promote the early embryonic development. PMID- 12575400 TI - [Effect of aloe-emodin on c-myc gene expression of cultured vascular smooth muscle cells of injured artery]. AB - OBJECTIVE: To study the molecular mechanism of aloe-emodin (AE) in inhibiting smooth muscle cells' (SMC) proliferation. METHODS: Deendothelialization was performed in abdominal aorta of Japanese white ear rabbits by using 3F Fogarty arterial embolectomy catheter. The media of abdominal aorta was isolated 48 hrs later for performing primary SMC culture. Cells were synchronized with G0/G1 phase by serum starvation, the AE was then added to the culture medium of experimental group, at a concentration of 20 micrograms/ml, and to the control group, equal volume of culture solution was added instead. The c-myc mRNA and protein expressions were examined 3 hrs later by using techniques of Northern blotting, Western blotting and immunocytochemistry respectively. RESULTS: In comparing with the control group, neither the expression of c-myc mRNA nor the expression of c-myc protein was changed after addition of AE. CONCLUSION: The inhibition of AE on SMC might not be due to influencing c-myc expression, but via other pathway. PMID- 12575401 TI - [Study on syndrome pattern in insulin resistant model rats]. AB - OBJECTIVE: To explore the Syndrome pattern in insulin resistant model rats. METHODS: Eight Sprague-Dawley (SD) rats were induced to insulin resistance (IR) by 60% high-sucrose forage. RESULTS: The correlative co-efficient of some biological and internal changes were clusterly analyzed and divided into 3 groups, which was closely related with phleg tubidity, blood stasis and internal toxin types respectively. The phlegm turbidity type was characterized by high content of blood lipid (triglyceride, total cholesterol) and high concentration of glycated serum protein; the blood stasis type was characterized by tendency of high viscosity and high coagulant state, the blood pressure increased, prothrombin time shortened, fibrinogen content raised and RBC and platelet count increased; while the internal toxin type was characterized by high content of glucose, insulin and the elevation of tumor necrosis factor. CONCLUSION: The combined Syndrome of phlegm turbidity, blood stasis and internal toxin is the Syndrome-pattern in insulin resistant model rat. This observation provides theoretic basis for clinical and experimental studies of TCM. PMID- 12575402 TI - [Serological study on inhibitory function of shenkang injection on glomerular mesangial cell]. AB - OBJECTIVE: To explore the intensity of inhibitory function of Shenkang injection (SKI), a Chinese herbal recipe, and benazepril on the proliferation of human glomerular mesangial cell (MC). METHODS: The effect of SKI and benazepril on the proliferation of human MC was observed by using the cultured MC in vitro and serum pharmacological assay. RESULTS: Both SKI and benazepril could inhibit the proliferation of human MC, but in equal dosage consistency, SKI showed a better inhibitory effect than that of benazepril. CONCLUSION: MC is the important target cell for the action of SKI, the inhibiting on proliferation might be one of the essential mechanisms of SKI in retarding glomerulosclerosis. PMID- 12575403 TI - [Discussion on rational medication of combination of Chinese herbal and Western medicine]. PMID- 12575404 TI - [The real significance of multiple targets effect in pharmacologic study of Chinese composite recipe should be stressed]. PMID- 12575405 TI - [Current status on establishment of local cerebral ischemic model]. PMID- 12575406 TI - [Review on studies of external treatment of skin ulceration with Chinese herbal medicine]. PMID- 12575407 TI - [Two cases successfully treated with long-term administration of low-dosage azithromycin]. PMID- 12575408 TI - [Continuous low-dosage macrolide therapy in children with bronchial asthma]. PMID- 12575409 TI - [Case of pulmonary MALT lymphoma successfully treated with clarithromycin]. PMID- 12575410 TI - [Stimulating effect of roxithromycin on biosynthesis of endogenous corticosterone]. PMID- 12575411 TI - [Effect of clarithromycin on lipopolysaccharide-induced anorexia]. PMID- 12575412 TI - [Efficacy of macrolide therapy for children with serous otitis media]. PMID- 12575413 TI - [Evidence-based macrolide therapy for children with serous otitis media]. PMID- 12575414 TI - [Effects of FK506 on neuroendocrine system--special reference to the effects on growth hormone secretory systems]. PMID- 12575415 TI - [Effects of 14-membered ring macrolide on transcription factors in human bronchial epithelial cells]. PMID- 12575416 TI - [Inhibitory effect of clarithromycin on NF-kappaB activation]. PMID- 12575417 TI - [Effects of clarithromycin on cytokine expression and NF-kappaB activation in nasal-mucosa derived cultured cells]. PMID- 12575418 TI - [Effect of macrolide antibiotics on MUC5AC production in human bronchial epithelial cells]. PMID- 12575419 TI - [Effect of macrolide antibiotics on mucus secretion from cultured epithelial cells of nasal mucosa]. PMID- 12575420 TI - [Effects of erythromycin and clarithromycin on chloride channels in bronchial epithelial cells]. PMID- 12575421 TI - [Experiment to establish the therapeutic dosage and administration term of macrolide antibiotics using the Explant Culture Method for human nasal mucosa]. PMID- 12575422 TI - [Effects of macrolide antibiotics on LPS-induced neutrophil and macrophage activation]. PMID- 12575423 TI - [Effects of roxithromycin on proliferation and activation of mast cells]. PMID- 12575424 TI - [Inhibitory effect of roxithromycin, a macrolide antibiotic, on Th2 cytokine production]. PMID- 12575425 TI - [Effect of azithromycin on cytokine production by THP-1 cells]. PMID- 12575426 TI - [Effect of azithromycin on cytokine production in human bronchial epithelial cell line, BEAS-2B]. PMID- 12575427 TI - Azithromycin reduces the production of virulence factors in Pseudomonas aeruginosa by inhibiting quorum sensing. PMID- 12575428 TI - [Suppressive activity of a macrolide antibiotic, roxithromycin, and its metabolites on co-stimulatory molecule expression]. PMID- 12575429 TI - [Mechanism in the cytoplasm accumulation of TOP binding factor]. PMID- 12575430 TI - [Effect of 14-membered ring macrolides on heparanase mRNA expression in lung cancer cells]. PMID- 12575431 TI - [Effect of macrolide therapy on microbes in nasal cavity and larynx of patients with chronic paranasal sinusitis]. PMID- 12575432 TI - [Anti-microbial sensitivities of bacteria isolated from patients with chronic lower respiratory tract infection under long-term macrolide therapy]. PMID- 12575433 TI - [MIC, sub-MIC of macrolide resistant bacteria--special reference to their quorum sensing system]. PMID- 12575434 TI - [Medical radiation physics of the "Wurzburg School"]. PMID- 12575435 TI - Extracranial stereotactic radiotherapy: evaluation of PTV coverage and dose conformity. AB - During the past few years the concept of cranial stereotactic radiotherapy has been successfully extended to extracranial tumoral targets. In our department, hypofractionated treatment of tumours in lung, liver, abdomen, and pelvis is performed in the Stereotactic Body Frame (ELEKTA Instrument AB) since 1997. We present the evaluation of 63 consecutively treated targets (22 lung, 21 liver, 20 abdomen/pelvis) in 58 patients with respect to dose coverage of the planning target volume (PTV) as well as conformity of the dose distribution. The mean PTV coverage was found to be 96.3% +/- 2.3% (lung), 95.0% +/- 4.5% (liver), and 92.1% +/- 5.2% (abdomen/pelvis). For the so-called conformation number we obtained values of 0.73 +/- 0.09 (lung), 0.77 +/- 0.10 (liver), and 0.70 +/- 0.08 (abdomen/pelvis). The results show that highly conformal treatment techniques can be applied also in extracranial stereotactic radiotherapy. This is primarily due to the relatively simple geometrical shape of most of the targets. Especially lung and liver targets turned out to be approximately spherically/cylindrically shaped, so that the dose distribution can be easily tailored by rotational fields. PMID- 12575436 TI - Homogeneous Ir-192 afterloading-flab-irradiation of plane surfaces. AB - Homogeneous irradiation of plane targets by Ir-192 afterloading flabs made by a parallel series of linear applicators can be time-consuming even with modern planning systems. The aim of the present study was to develop an algorithm that supplies homogeneous dose distributions in an arbitrary given plane in parallel to the equipped plane of a flab. The edge and corner positions of the flab are of particular importance. The identity of the dose in the optimisation distance above the flab centre, corners, and middle of the flab edges, leads to a strict relation of the respective dwell weights. Formulas can be derived that allow the calculation of the dwell times. The dimensioning of the flab can be rapidly adapted to new conditions. A comparison with the results of Nucletron PLATO-BPS for applicator-applicator distances and step sizes of 1 cm at optimisation distances of 10, 20, 30, and 40 mm and various flab sizes (3 x 3, 9 x 9, and 15 x 15 cm2) shows the following results: The standard deviation of the proposed algorithm is sometimes slightly higher than the results of the commercial planning system, whereas the underdosage at the flab edges is usually smaller. The effort for planning and preparation of the irradiation, for example using a Nucletron HDR, is below 5 minutes--a considerable reduction of planning time. PMID- 12575437 TI - CT treatment planning based on a simulator CT-option: comparison between calculation and measurements using TLDs in a humanoid phantom. AB - Computer programs are widely used in radiotherapy to calculate the dose distribution in patients for different field configurations. The aim of this study was to investigate the dosimetry over the complete radiotherapy procedure. For this purpose, CT slices of a humanoid phantom (Alderson) were first taken with a simulator CT-option and the images then transferred to the treatment planning system. Irradiations were performed using linear accelerators. Measurements in the phantom were carried out with thermoluminescence dosimeters (TLD) and analysed using an automatic reader. Several common treatment configurations were investigated. Measurements within the target volume (low dose gradient) agreed with the calculation to within 3%. The differences in the penumbra region (high dose gradient) were in part much larger. This can be attributed to two factors: the uncertainties in the TLD localisation and the calculation chain. In conclusion, it can be stated that the agreement between calculations and measurements was sufficient for most clinical considerations. However, further checking is advisable in case sensitive organs lie in the vicinity of the field edge. PMID- 12575438 TI - A computer aided diagnostic system for radiotherapy planning. AB - Planning for radiation therapy intervention implies the definition of treatment volumes as well as a clear delimitation of normal tissue. This paper presents a Computer Aided Diagnostic system for the automatic CT image analysis. Two important problems are solved: the spinal cord segmentation and the detection of lung metastases. Some subordinate problems are also solved: the detection of spinal canal, lamina, lungs, and ribs, as well as the identification of thorax contour. The developed methodologies use a knowledge-driven image processing based on Anatomical Structures Maps and task-oriented architecture. Experiments were performed on CT images from La Chaux de Fonds Hospital (Switzerland). Evaluations were performed using a visual inspection of the contours projected on the CT image slices. The radiologist decided whether each of the contours obtained with our system was acceptable or not. The accuracy of the method was defined as the fraction of CT slices in which the particular contour was correctly located. In the case of spinal cord segmentation, the procedure was tested on 23 patients (1051 images), resulting in an accuracy of 91%. In the case of lung tumors detection, the method showed an accuracy of > 90%, with testing performed on 20 patients for a total of 988 images. The experiments performed show that the method is reliable, with possible future application in an oncology department. PMID- 12575439 TI - Dosimetric calculations and VIPAR polymer gel dosimetry close to the microSelectron HDR. AB - In the present study, different dosimetric methods were investigated for their ability to predict the energy dose in the vicinity of the microSelectron HDR 192Ir brachytherapy source. The results of a time-efficient Sievert integral model of proven accuracy in the cm distance range from all 192Ir sources were benchmarked against accurate Monte Carlo derived dosimetric data in the close vicinity of the source. This comparison revealed that the Sievert model is capable of accurate dosimetry even in the mm distance range from the source. The dose rate distributions were compared with results obtained from different versions (v. 13.7 and 14.2.2) of the Plato BPS commercial treatment planning system, for an application following the Paris trial intravascular irradiation protocol. The results of brachytherapy planning system calculations were found reliable at radial distances of clinical relevance. Noticeable errors existed only in the extreme case of dose calculations at 2 mm from the source axis using Plato v. 13.7. Experimental dosimetric data for the intravascular application, as obtained through the VIPAR polymer gel-MRI method, were also evaluated for dose verification purposes. This method allowed with reasonable accuracy the verification of absolute dose distributions for peripheral vessel applications using 192Ir sources. PMID- 12575440 TI - [Long term registration of linear accelerator parameters for quality assurance in stereotaxic radiosurgery]. AB - Long-term observations of mechanical parameters have been performed in Freiburg for the last 7 years within the quality assurance of stereotactic irradiation at linear accelerators. The deviations between the laser indication and the position of the beam isocentre are recorded, as well as parameters for the stability of the couch rotation and the additional devices for patient adjustment and beam collimation. The deviations are used for the correction of the calculated isocentre coordinates. The long-term observation of the measured values and their standard deviations allow conclusions about the quality of the measuring procedure itself, the laser adjustment, the extension of the isocentre sphere, as well as the mechanical slackness of the treatment couch and the fixation device of the stereotactic ring. The standard deviations of the isocentre position were within 0.3 to 0.8 mm, those of the measurement quality approximately 0.2 mm. The analysis of long-term observation has contributed to improvements of the equipment and of quality assurance procedures. PMID- 12575441 TI - [Acceptance and constancy test radiographic projections with screen film systems]. AB - The relation of dose and optical density in a conventional X-ray unit working with Screen Film Systems is subject to inherent fluctuations. For conventional equipment, the fluctuations of the components in conformity to the manufacturer instructions are calculated in such a fashion that the X-ray images of the patients fullfil the requirements of the Radiological Guidelines. The tolerance requirements of a mammogram can only be maintained if the operator runs the subsystem of film and processing such that the processing of films with single emulsion number remains over a longer period of time as close as possible to the manufacturer specifications for the type of film. The testing strategy used thus far in acceptance tests is the random sampling. In conventional X-ray units, and within the frame of constancy tests, this leads to an insufficiently accurate conclusion in case of exceeding tolerances of test-phantom images: of 99 images, 9 give a wrong result (rate of false negative + false positive). In mammography units, the introduction of stricter tolerances (delta D = +/- 0.15; desirable according to the "European Protocol for the Quality Control of the Physical and Technical Aspects of mammography Screening"), would further decrease this ratio: 37 of 99 images would give a wrong result. This forces to a different strategy of testing when the reference values are fixed for the image of a test phantom and for the constancy testing of film processing. PMID- 12575442 TI - [Comments on "Dosimetric verification of intensity-modulated radiotherapy in the German Cancer center in Heidelberg"]. PMID- 12575443 TI - [Structure of genomic domains of mammalian and avian globin genes]. AB - The data on the genomic domain structure of both mammalian and avian alpha- and beta-globin genes are reviewed. The specific features of chromatin, DNA binding to the nuclear matrix, and domain-specific transcripts are discussed. In humans, the beta-globin gene domain is located in the GC-depleted isochore and contains multiple nuclear matrix attachment regions. The locus is controlled by six chromatin regions hypersensitive to DNase located far upstream of the first structural gene. Some of these regions display enhancer activity to support normal transcription level in the domain. Other mammalian beta-globin domains are similarly organized. The avian beta-globin genes are specifically arranged and their expression is less dependent from the locus control region. The human alpha globin gene is located in the GC-rich isochore. The nuclear matrix attachment sites are not identified in this gene. An analog of the locus control region is located 40 kb upstream of the zeta-globin gene. The avian alpha-globin gene domains contain numerous nuclear matrix attachment regions. In these domains, an element located far upstream the genes regulates positive rather than negative transcription. An unidentified housekeeping gene as well as some other transcripts not encoding the structural globin genes is transcribed in the direction opposite to that of the globin genes in both mammalian and avian domains. PMID- 12575444 TI - [Monomers of a satellite sequence of chaffinch (Fringilla coelebs L., Aves: Passeriformes) genome contains short clusters of the TTTAGGG repeat]. AB - A novel repeated sequence of chaffinch (Fringilla coelebs) designated as GS was isolated from genomic DNA after in vitro amplification of satellite DNA sequences using GSP-PCR technique. The proportion of this repeat in the chaffinch genome constitutes about 2%. Monomers are 176 to 199 bp in size and contain a short cluster of the TTAGGG telomeric tandem repeat. The oligomer of the telomeric hexanucleotide is flanked by the sequences that are significantly different in different monomers. The GS sequences are organized as tandemly repeated units and located in a number of chromomycin-positive blocks on the long arms of macrochromosomes 1, 2, 3, 5, and 6, as well as on several microchromosomes. The sequences homologous to the GS satellite of chaffinch were not found in the genomes of redwing (Turdus iliacus) and house sparrow (Passer domesticus). PMID- 12575445 TI - [Identification and cloning of peptide synthetase genes of thermostable bacilli using the polymerase chain reaction]. AB - Fourteen thermophilic and thermostable strains of the genus Bacillus were studied. Total DNA was isolated from these strains and used as a template to identify and clone peptide synthetase genes by means of polymerase chain reaction. Amplified DNA fragments were cloned into a phasmid vector, and nucleotide sequences of cloned fragments were determined. Stringent thermophilic strains were shown to lack genetic systems, which are responsible for the synthesis of secondary metabolites and homologous to the known peptide synthetase genes. On the contrary, thermostable strains had peptide synthetases and produced antimicrobial secondary metabolites. Analysis of nucleotide sequences and deduced amino acid sequences of cloned PCR fragments from B. licheniformis strains VK2, VK21, and VK2101 showed that they are absolutely identical. The cloned DNA fragment was found to be a portion of the open reading frame, which we termed ORF1. Data from analysis of a partial nucleotide sequence of the peptide synthetase gene of strain VK21 indicated that a 9.5-kb region of chromosomal DNA contains sequences of two genes homologous to the B. subtilis peptide synthetase genes dhbB and dhbF. Strains VK2, VK21, and VK2101 were shown to synthesize siderophores. A method for screening bacteria with peptide synthetase genes has been developed. PMID- 12575446 TI - [The effect of the genotype on the expressiveness of the vestigial trait and polyteny of chromosomes in Drosophila melanogaster Meig]. AB - The effects of the Bar (B) and white (w) mutations on the expressiveness of the character vestigial (vg) and the degree of polyteny of salivary gland giant chromosomes were studied in Drosophila melanogaster. Either mutation changed both the expressiveness of vestigial and the degree of chromsome polyteny. A negative association between the vg expressiveness and the degree of chromosome polyteny was revealed and proved to be stronger in females than in males. The parameters under study were shown to differ between females and males. PMID- 12575447 TI - [Cytogenetic analysis of two ecdysone-regulated puffs 62C and 62E in Drosophila melanogaster]. AB - Genetic analysis has been performed to reveal vital genes around two puffs, a late 62C puff and an early-late 62E puff. Their roles in hormonal regulatory mechanisms have been estimated. A locus represented by four lethal mutations has been found in the vicinity of the 62E puff. The mutants display disturbed puffing, which suggests the involvement of this locus in hormonal regulatory mechanisms. In the 62C puff region. 26 mutations have been found that proved to be allelic to mutations in the D-Titin gene. The giant D-Titin gene is essential for the sarcomeric organization of striated muscles. According to the results of in situ hybridization with polytene chromosomes, the D-Titin gene occupies the entire 62C puff. The phenotypic characteristics of the novel mutants suggest that this protein is polyfunctional, and its role is not restricted to processes in the muscular tissue. It may also be involved in the morphogenesis of leg imaginal disks, and it is necessary for condensation and separation of sister chromatids during mitosis. Mutations in the ecdysone-induced BR-C and E74 genes cause disturbances similar to those found in this study. In addition, mutations of these genes can affect the D-Titin gene activity, which suggests that the three genes are involved in similar morphogenetic and myogenetic processes. PMID- 12575448 TI - [About the possibility of hybridogenesis in the species origin of the midge Chironomus usenicus Loginova et Beljanina (Chironomidae, Diptera)]. AB - Hybridogenesis as a possible way of speciation in Chironomidae was considered with special reference to the species Chironomus usenicus resulting from hybridization between C. plumosus and C. behningi. The three species had 2n = 8 and belonged to the thummi cytocomplex with chromosome arm combinations AB, CD, EF, and G. Arm G had a marker chromosome disk sequence (CDS) and was used to demonstrate the hybrid origin of C. usenicus. Most C. usenicus larvae were heterozygous in CDS of arm G. CDS use G2 proved to be identical to CDS beh G1 of C. behningi and CDS use G1, to CDS plu G1 of C. plumosus. It was assumed that C. usenicus results from hybridization between eurybiont C. plumosus and stenobiont C. behnigni at the boundary of their species areas, in freshwater or brackish water bodies of the southern Saratov oblast and northern Kazakhstan. Morphologically and karyotypically, the hybrid was probably similar to C. plumosus. Crosses with C. plumosus eliminated virtually all C. behningi chromosome sequences from the karyotype of the hybrid. Further chromosome divergence resulting in C. usenicus involved a number of chromosome rearrangements, including duplication of pericentric heterochromatin and other chromosome regions; inversion, which occurred in arm F (regions 13-16) and was fixed in the karyotype; and other paracentric inversions and deletions accumulated in heterozygote in the karyotype pool of the species. Since C. behningi was eliminated from the introgression zone and its species are reduced, the assimilation character was assumed for introgressive hybridization of C. behningi and C. plumosus. PMID- 12575449 TI - [Genetic variability in Scotch pine in the southeastern part of its range]. AB - The main parameters of genetic variability have been determined in an isolated natural Scotch pine population from Chita oblast' (Tsasuchei Forest) by analysis of 19 genes coding for nine enzymes: GDH, IDH, LAP, PGM, AAT, ADH, MDH, 6-PGD, and DIA. Polymorphic genes constituted 63.2% of all structural genes studied in the population at the 99% polymorphism criterion. The mean number of alleles per locus was 1.63. The observed and expected heterozygosities were 0.237 and 0.251, respectively. These estimates are close to the corresponding mean values for Scotch pine according to the data on 18 or more structural genes. PMID- 12575450 TI - [Molecular analysis of leaf-rust resistant introgression lines obtained by crossing hexaploid wheat Triticum aestivum with tetraploid wheat Triticum timopheevii]. AB - Twenty-four Triticum eastivum x T. timopheevii hybrid lines developed on the basis of five varieties of common wheat and resistant to leaf rust were analyzed by the use of microsatellite markers specific for hexaploid common wheat T. aestivum. Investigation of intervarietal polymorphism of the markers showed that the number of alleles per locus ranged from 1 to 4, depending on the marker (2.5 on average). In T. timopheevii, amplification fragments are produced by 80, 55, and 30% of primers specific to the A, B, and D common wheat genomes, respectively. Microsatellite analysis revealed two major areas of introgression of the T. timopheevii genome: chromosomes of homoeological groups 2 and 5. Translocations were detected in the 2A and 2B chromosomes simultaneously in 11 lines of 24. The length of the translocated fragment in the 2B chromosome was virtually identical in all hybrid lines and did not depend on the parental wheat variety. In 15 lines developed on the basis of the Saratovskaya 29, Irtyshanka, and Tselinnaya 20, changes occurred in the telomeric region of the long arm of the 5A chromosome. Analysis with markers specific to the D genome suggested that introgressions of the T. timopheevii genome occurred in chromosomes of the D genome. However, the location of these markers on T. timopheevii chromosomes is unknown. Our data suggest that the genes for leaf-rust resistance transferred from T. timopheevii to T. aestivum are located chromosomes of homoeological group 2. PMID- 12575451 TI - [A virulence gene from Xanthomonas campestris pv. campestris homologous to the avrBs2 locus is recognized in race-specific reaction by two different resistance genes in Brassica plant species]. AB - Race-specific interaction between the Brassica plants and Xanthomonas compestris pv. campestris bacteria follows the "gene-for-gene" rule. Expression of the avirulence genes recognized by two dominant resistance genes of Brassica, Rxc1 in plants with the BB genome, and Rxc3 in the CC plants, was lost after bacterial mutation in planta. The mutation was distinguished by the elongation of CGCGC pentanucleotide repeat in the gene, which was designated as avrRxc1/3. This gene displayed strong structural similarity to the avrBs2 locus from the related species X. vesicatoria. Thus, it is the first description of the avrRxc1/3 avirulence gene conferring race-specific interaction between X. campestris pv. campestris and Brassica plants. Structural homologues of the avrBs2 are found in many Xanthomonas species, but in all cases except X. vesicatoria, their function remains unknown. PMID- 12575452 TI - [Breeding for digenic apple resistance to scab]. AB - The results of the apple breeding for digenic resistance to scab (1979-2000), which is more long-term than the monogenic breeding, have been reviewed. The hybrid seeds obtained from the reciprocal crossing between the Vf and Vm resistance gene donors served as the original material. The seed progeny yielded by backcrosses between these hybrids and the susceptible cultivars were examined (age--first true leaves) using inoculation in a greenhouse. The following criteria were proposed for breeding parental forms and the cultivars of the VfvfVmvm genotype: (1) phenotype segregation in the progeny of three resistant: one susceptible seedling and (2) the presence in the progeny of two types of resistant seedlings: those with the class 1 resistance (points and spots hypersensitivity) typical of the Vm gene and those with the class 2-3 resistance (chlorotic and necrotic spots) typical of the Vf gene. PMID- 12575453 TI - [Genetic diversity of protein markers in sheep population from Ukraine]. AB - Based on polymorphism of genes for antigen factors of six blood-group systems and four blood protein loci, genetic structure and the main variation parameters were studied in three sheep breeds and three sheep breed types constituting the basis of purebred sheep resources in Ukraine. Specific features of the distribution of genotypes and alleles of polymorphic loci were determined in each of the studied sheep groups depending on their origin and production type. The molecular-genetic markers used in the analysis of the genetic relationships between the sheep breeds and breed types were shown to objectively reflect their breeding history and evolution. Integrally, each of the studied gene pools had a specific profile of gene frequencies reflecting breeding specificity, breed history, and genetic differentiation of breeds. PMID- 12575454 TI - [Specificity of gene di (diabetes insipidus) expression in homologous inbred rat strains]. AB - The diabetes insipidus mutation is displayed in homozygotes in the form of diabetes insipidus with water consumption from 30 to 100% of body weight per day. We developed two inbred sublines of the di/di Brattleboro rats as well as the recombinant inbred subline by integrating genes of August rats into the di/di mutant genome. Changes in the genetic background proved to have no effect on the quantitative parameters of the diabetes insipidus. The intensity of the secondary immune response and the content of tropomyosin in the medulla of the rat kidney can serve as additional marker traits of the di/di genotype. PMID- 12575455 TI - [The genotoxic effect of gunshot wounds]. AB - Evidence for alterations in chromosomes of experimental animals (rats) and humans after gunshot wounds is presented. The rate of chromatid exchanges induced by gunshot wounds in humans depend on the saturation of body tissues with ascorbic acid. It is assumed that free-radical processes underlie the deleterious effect of gunshot wounds on chromosomes. PMID- 12575456 TI - [Detection of aneuploidy in spontaneous abortions using the comparative hybridization method]. AB - Comparative genomic hybridization (CGH) technique was used to examine a set of ten spontaneous abortions whose cell cultures were characterized by the lack of proliferation in vitro, and thereby, were not available for the analysis by means of routine cytogenetic methods. Five abortions (50%) had aneuploidy of autosomes, including trisomy 10, 14, 18, and 21, and monosomy 22. The latter variant of unbalanced chromosomal abnormalities is rarely detected in spontaneous abortions by use of conventional cytogenetic methods. The results were validated by using fluorescent in situ hybridization (FISH) analysis with centromere-specific DNA probes. Embryos with trisomy 10 and monosomy 22 displayed mosaicism with the frequencies of abnormal cell clones constituting 68 and 33% respectively. The advantages and limitations of the applying of CGH technique for detection of genomic abnormalities in both nonmosaic and mosaic forms are discussed. PMID- 12575457 TI - [Analysis of association of Col1a1 gene alleles with the development of osteoporosis]. AB - Allele frequencies of the G-->T polymorphism at the regulatory region of the Collal gene in the population of the northwestern Russia (control group) and in osteoporotic patients were estimated by the RFLP method based on PCR-mediated site-directed mutagenesis. Three patient groups with radiologically confirmed osteoporosis were examined. Group 1 consisted of 64 patients with severe osteoporosis complicated by fractures (SO); group 2 included 15 children with idiopathic osteoporosis (IO); group 3 consisted of 98 women with postmenopausal osteoporosis developed at the background of estradiol-deficiency state (PMO). The frequency of functionally defective allele s in the control group was 16.7%. It was statistically different from that in the SO patients (48.4%) (P < 0.01) and in the IO children (40%) (P < 0.01). The frequency of allele s in the PMO patients constituted 23.0% and it was similar to that in the control group (P > 0.05). Analysis of the Collal alleles provides early detection of the individuals with hereditary predisposition to osteoporosis and prophylaxis of the disease at the presymptomatic stage. PMID- 12575458 TI - [Molecular genetic analysis of essential tremor]. AB - Essential tremor (ET) is the most common extrapyramidal disorder of the central nervous system with autosomal dominant transmission in the majority of cases and age-dependent penetrance of the mutant gene. In a number of cases, it shares some phenotypic features with autosomal dominant idiopathic torsion dystonia (locus DYT1 on chromosome 9q32-34) and is genetically heterogeneous: distinct variants of ET were mapped to chromosomes 3q13 (ETM1) and 2p22-25 (ETM2). We performed studies of candidate loci in a group of Slavonic (11 patients) and Tajik (19 patients) families with ET. Mutational analysis of the DYT gene in probands did not reveal the major deletion 946-948delGAG characteristic of idiopathic torsion dystonia, which allows one to genetically distinguish the studied hereditary forms of ET and torsion dystonia. Based on analysis of genetic linkage in informative Tajik pedigrees with ET, linkage to locus ETM1 on chromosome 3q13 was established in four families. Maximum pairwise Lod score was 2.46 at recombination fraction of theta = 0.00; maximum combined multipoint Lod score was 3.35 for marker D3S3720 and a common "mutant" haplotype for markers D3S3620, D3S3576, and D3S3720 allowed us to locate a mutant gene in a relatively narrow chromosome region spanning 2 cM. In one informative pedigree with ET, both candidate loci ETM1 and ETM2 were definitely excluded on the basis of negative Lod scores obtained by linkage estimations, which testifies to the existence of another distinct gene for autosomal dominant ET. PMID- 12575459 TI - [Polymorphism of interleukins and interleukin receptor genes: population distribution and association with atopic bronchial asthma]. AB - Population distribution and pathogenetic significance for bronchial asthma (BA) of the eight polymorphic variants of six interleukin--(IL) and interleukin receptor genes, C-589T, G/C 3'-UTR IL4, C-703T IL5, T113M IL9, Q551R, 150V IL4RA, and G1972A IL5RB, was examined. In the population samples of Russians, Tajiks, Buryats, and Tuvinians racial and ethnic specificity of these polymorphisms was established. These specific features were manifested as population-specific "enetic portraits" in respect of polymorphic allele frequencies. Analysis of the BA patients and their relatives from Tomsk by use of transmission/disequilibrium test (TDT) revealed the presence of a statistically significant association between the C-703 IL5 allele and the disease (P = 0.005). This is the first evidence of an association between the IL5 gene polymorphism and BA. PMID- 12575460 TI - [The ISBsu2 mobile element is present in a plasmid of a soil strain and in the chromosomes of several other strains of Bacillus subtilis]. AB - Chromosomes of several Bacillus subtilis strains were shown to contain homologs of the ISBsu2 mobile genetic element, which was earlier revealed in a cryptic plasmid of a soil strain of B. subtilis. PMID- 12575461 TI - [A simple and rapid method for detection of the 106Gln mutation in Wilson Konovalov disease]. AB - A simple and rapid method for detecting the 1069Gln mutation in gene ATP7B based on a PCR specific for this allele has been developed. The 1069Gln mutation is the main cause of Wilson disease (WD) in Russia and accounts for approximately 40% of all mutant alleles of gene ATP7B. Therefore, the method proposed makes the postnatal and prenatal diagnosis of Wilson disease in Russia considerably more rapid and less expensive. PMID- 12575462 TI - Pharmaco-EEG: from cellular to the network level. EEG beta spectral power as a biomarker for GABAergic inhibition and its role in the assessment of AEDs. PMID- 12575463 TI - Standardized low-resolution brain electromagnetic tomography (sLORETA): technical details. AB - Scalp electric potentials (electroencephalograms) and extracranial magnetic fields (magnetoencephalograms) are due to the primary (impressed) current density distribution that arises from neuronal postsynaptic processes. A solution to the inverse problem--the computation of images of electric neuronal activity based on extracranial measurements--would provide important information on the time-course and localization of brain function. In general, there is no unique solution to this problem. In particular, an instantaneous, distributed, discrete, linear solution capable of exact localization of point sources is of great interest, since the principles of linearity and superposition would guarantee its trustworthiness as a functional imaging method, given that brain activity occurs in the form of a finite number of distributed hot spots. Despite all previous efforts, linear solutions, at best, produced images with systematic nonzero localization errors. A solution reported here yields images of standardized current density with zero localization error. The purpose of this paper is to present the technical details of the method, allowing researchers to test, check, reproduce and validate the new method. PMID- 12575465 TI - Genetic programming approach for the optimal selection of combinations of neuronal networks to classify sleep stages by QUISI. AB - The usefulness of a new way to optimize the cooperation of trained neural networks for automatic one-channel sleep stage analysis using genetic programming and performance evaluation by including the interrater reliability are the focus of our paper. The one-channel sleep classification could be significantly improved by the optimization. The software tool HENNE, with its genetic programming compartment was developed for this purpose. The tool has proved to be useful for searching for optima in difficult goal surfaces. To contribute to the general discussion about the benefit of the automatic one-channel sleep analysis on the basis of the frontal site, we tried to evaluate our results before the background of the interrater variability. Comparing the kappa statistics of different independent studies with our results, we concluded that there are no dramatic differences as a rule and that QUISI is a useful device as a presleep laboratory and ambulatory diagnostic tool. PMID- 12575464 TI - Simultaneous ERP and event-related fMRI: focus on the time course of brain activity in target detection. AB - The event-related P300 potential has been widely used in neurophysiological research. It is usually evoked with an oddball paradigm. One main reason for its broad application in neurophysiological research is the fact that in several brain/mental diseases, such as Alzheimer's disease or schizophrenia, attenuations of the P300 amplitude and latency have been described. However, a precise correlation of the scalp data to the underlying brain regions was not possible, as the correct localization of the generators of scalp-measured electroencephalogram (EEG) data was limited, due to the low spatial resolution of EEG-data. With the availability of modern imaging technologies, functional Magnetic Resonance Imaging (fMRI) in particular, the underlying brain activations could be detected using an oddball task. Although the spatial resolution of fMRI is excellent, the time resolution is restricted. For a comprehensive understanding of the brain activity underlying the P300 paradigm, we have used a combination of EEG and fMRI to get a precise localization and a high-time resolution of the underlying brain activity. PMID- 12575466 TI - The automatic recognition of REM sleep: a challenge and some answers. AB - Automatic sleep staging has remained a constant challenge over the years due to scientific and economic reasons. Rapid eye movement (REM) sleep recognition is a most critical subtask for these procedures. Because the physiological meaning of REM sleep is still an open question and the identification depends on the coordinated behavior of three biological signals, the present definition still has to be considered preliminary and arbitrary to some extent. Three neural network-based algorithms for automatic REM sleep recognition are reviewed herein. All of them rely on a single EEG channel as input signal, but differ in the preprocessing method. The results are encouraging but might be improved with respect to the interrater concordance rate. Applying nonlinear measures could not reduce the errors in our study. Other biological parameters are discussed that might help to improve the results. PMID- 12575467 TI - The key-lock principle in the diagnosis and treatment of nonorganic insomnia related to psychiatric disorders: sleep laboratory investigations. AB - Nonorganic insomnia is a frequent sleep disorder that has a high comorbidity with other psychiatric illnesses. In our sleep outpatient clinic, 41% of the patients showed neurotic, stress-related and somatoform disorders, 31% affective disorders and 1.6% schizophrenia. Sleep laboratory investigations in patients for diagnostic purposes and in normal subjects for the evaluation of drug effects suggest that changes in the sleep architecture of patients with nonorganic insomnia due to psychiatric disorders, compared with normal controls, are opposite to alterations induced by psychotropic drugs intended for their treatment, compared with placebo (key-lock principle). Evidence for this principle was found regarding nonorganic insomnia related to generalized anxiety disorder or panic disorders and benzodiazepines, depressive episodes, recurrent depression or dysthymia and sedative antidepressants and finally schizophrenia and sedative neuroleptics. Polysomnography (PSG) findings of other mental disorders are rather scarce and often depend upon the subtype and stage of the disease. In conclusion, sleep laboratory studies may be helpful for choosing the right drug for an individual insomniac patient. PMID- 12575468 TI - Unsupervised continuous sleep analysis. AB - One aim of the EU-funded project SIESTA was to develop a new way of describing the human sleep-wake continuum with high temporal resolution, and independent of subjective rules, to serve as an alternative to traditional sleep scoring. Here, we report new findings obtained with a fully automatic, probabilistic sleep analyzer using Hidden Markov Models (HMMs) based on data from a single electroencephalogram (EEG) channel. HMMs allow the analysis of non-stationary time series by modeling both the probability density functions of locally stationary data and the transition probabilities between these stable states. In the context of sleep analysis, the locally stable states can be thought of as sleep stages. The sleep-wake continuum was modeled as a mixture of three different processes by defining a three-state Gaussian Observation HMM (GOHMM). No class information from human scorers was used to train the model. The probabilities of being in any of the three states at each point in time roughly indicate the amount of wakefulness, deep sleep and rapid-eye-movement (REM) sleep with a one-second time resolution. Although it was not the aim to replicate the traditional Rechtschaffen and Kales (R&K) scoring, pseudo R&K hypnograms were constructed from the probability plots in order to compare the analyzer results with classical sleep stages by human experts. We expected that the analyzer would be able to classify data correctly from the "cornerstones" of human sleep (wakefulness, deep sleep, and REM sleep). Contrary to our previous efforts, we trained the HMMs on data from two different sleep laboratories separately, instead of generalizing data from diverse laboratories. While these stages could be detected with an accuracy of around 80% at the sleep laboratory for which we already had achieved the best results, there was no improvement from previous results by the training of a separate model in the other laboratory. This finding indicates clear laboratory effects in the signal characteristics, probably due to differences in hardware and filter settings. The presented approach, going beyond a mere replication of the traditional R&K standard, offers a continuous description of human sleep which is based on probabilistic principles. It provides a second-by-second quantification of the sleep-wake continuum and captures, although being entirely data-driven instead of rule-based, the three main processes in human sleep: wakefulness, deep sleep and REM sleep. PMID- 12575469 TI - Animal pharmaco-EEG: models, facts and fallacies. PMID- 12575470 TI - Electroencephalography parameters as biomarkers: extrapolation from laboratory animals to humans. PMID- 12575471 TI - Rat models of genetic absence epilepsy: what do EEG spike-wave discharges tell us about drug effects? AB - Electroencephalographic studies in the WAG/Rij rats of Nijmegen and genetic absence epileptic rats of Strasbourg (GAERS), two genetic models for human generalized absence epilepsy, illustrate the usefulness of drug electroencephalogram (EEG) interaction studies. In the EEG of both types of rats, spontaneously occurring spike-wave discharges are present. For drug discovery, a model with predictive validity is imperative, and both the WAG/Rij and the GAERS models seem adequate. The present paper discusses effects on spike-wave discharges of various compounds that are clinically used. Not only new antiepileptic drugs, such as remacemide, loreclezole, lamotrigine, tiagabine, gabapentin, progabide and levetiracetam are evaluated, but also drugs used for other purposes, such as etomidate and fentanyl-fluanisone for anesthesia, opioidergic drugs and drugs used for strokes. It is shown that some new antiepileptic drugs, such as tiagabine, have spike-wave discharge-increasing properties, while other drugs are worth studying in clinical trials for antiabsence treatment. Furthermore, it is shown that many commonly used drugs such as analgesics, anesthetics and drugs to treat stroke generally enhance spike wave discharges. It can be concluded that EEG monitoring is imperative for the discovery and development of potentially antiepileptic compounds and that genetic rat models such as the WAG/Rij or GAERS, to a large extent, can reliably predict clinical efficacy of various types of compounds as well as alert us of potentially adverse effects. PMID- 12575472 TI - Signature profiles in sleep-wake drug discovery. PMID- 12575473 TI - Identifying responders to serotonin agonists by neuroelectric brain activity. PMID- 12575474 TI - Structural and energetic processes related to P300: LORETA findings in depression and effects of antidepressant drugs. AB - Noninvasive electrophysiological neuroimaging applied to cognitive components of event-related potentials (ERPs) may differentiate between structural and energetic processes related to information processing. The structural level, revealed by the location of the local maxima of the current source density distribution, describes the time-dependent network of activated brain areas. The magnitude of the source strength, a measure of the energetic component, describes the allocation of processing resources. ERPs were recorded in an odd-ball paradigm and low-resolution brain electromagnetic tomography (LORETA) was applied for standard and target ERP components. In a group of 60 menopausal depressed patients of 45-60 years of age, reduced P300 source strength was observed bilaterally, temporally and medially prefrontally reaching to rostal parts of the anterior cingulate, compared with 29 age-matched controls. In a double-blind, placebo-controlled study, 2 mg of the antidepressant citalopram induced a significant increase of P300 source strength in the (left) prefrontal cortex and precuneus compared with placebo, reaching to the posterior cingulate. Similar increases were observed after 800 mg S-adenosyl-L-methionine (SAMe) administered intravenously in ten young healthy subjects aged 22-33, and they were even more pronounced in ten elderly healthy subjects aged 56-71. Thus, ERP-tomography identified changes in energetic sources in brain areas predominantly involved in depression and in antidepressant action. PMID- 12575475 TI - EEG topography and tomography in diagnosis and treatment of mental disorders: evidence for a key-lock principle. AB - Clinically well-defined diagnostic subgroups of mental disorders, such as schizophrenia with predominantly plus and minus symptomatology, major depression, generalized anxiety disorder, agoraphobia, obsessive-compulsive disorder, multiinfarct dementia, senile dementia of the Alzheimer type and alcohol dependence, show electroencephalogram (EEG) maps that differ statistically both from each other and from normal controls. Representative drugs of the main psychopharmacological classes, such as sedative and nonsedative neuroleptics and antidepressants, tranquilizers, hypnotics, psychostimulants and cognition enhancing drugs, induce significant and typical changes to normal human brain function compared with placebo, in which many variables are opposite to the above mentioned differences between psychiatric patients and normal controls. Thus, by considering these differences between psychotropic drugs and placebo in normal subjects, as well as between mental disorder patients and normal controls, it may be possible to choose the optimum drug for a specific patient according to a key lock principle, since the drug should normalize the deviant brain function. This is supported by low-resolution brain electromagnetic tomography (LORETA), which identifies brain regions affected by psychiatric disorders and psychotropic drugs. PMID- 12575476 TI - Spatial configuration of brain electric activity during positive, neutral and negative emotions. PMID- 12575477 TI - Cholinergic function and dysfunction in the visual system. AB - Acetylcholine (ACh) function is thought not only to play a significant role in memory, learning and other cognitive processes, but studies at a cellular level and in vivo indicate an important role for ACh in vision as well, especially for visual information processing. A suitable experimental model of geriatric memory impairment and Alzheimer dementia that pharmacologically blocks the brain muscarinic transmission has been proposed. This model has been extensively used also as an attempt to test cholinergic drugs in the absence of detailed knowledge of sites and mechanisms of ACh action and as test condition in the investigation of the role of ACh in visual information processing. Alzheimer's dementia results from complex neuron alterations, rather than simply reflecting ACh impoverishment, also involving the visual system, with substantial loss of retinal ganglion cells and alterations in visual information processing. Viewing all these data as a whole, nonspecific ACh actions on cognition, such as arousal or attention, contribute in modulating the function-specific action of ACh in information processing, both at cognitive and visual level. PMID- 12575478 TI - Sleep electroencephalogram alterations disclose initial stage of encephalopathy. AB - Minimal hepatic encephalopathy is often present in patients with chronic liver disease. The aim of this work was to determine changes in the dynamics of sleep electroencephalogram (EEG) in cirrhotic patients without overt encephalopathy. Twenty such cirrhotic subjects included in the protocol of hepatic transplantation of our hospital were studied and compared with 20 age-matched healthy volunteers. Spectral analysis of all-night EEG was estimated by computing the fast-Fourier transform in 2-second epochs, and averaging every 60 seconds. Artifacts were off-line suppressed, sleep stages (stage 2, stage 3-4 and REM) were determined, and the EEG mean dominant frequency (MDF) was calculated in each of these stages. Results show that in cirrhotic patients, nocturnal MDF evolution discloses a clear alteration of the ultradian EEG frequency oscillations present in controls. Also, the mean value of MDF in REM episodes was larger in cirrhotics than the corresponding value in controls. CONCLUSIONS: 1) sleep EEG evidences the existence of minimal hepatic encephalopathy; 2) the spectral analysis of EEG in minimal hepatic encephalopathy showed that the changes of MDF during sleep are an early marker of cerebral dysfunction in cirrhotic patients. PMID- 12575479 TI - Pharmacokinetic/pharmacodynamic relationships: basic concepts. PMID- 12575480 TI - Pharmaco-electroencephalography and pharmacokinetic/pharmacodynamic modeling in basic research: focus on human pharmacology. AB - Pharmacokinetic/pharmacodynamic modeling, together with electroencephalography (EEG), have been successfully applied to obtain in vivo pharmacological information of different drugs acting on the central nervous system (CNS) in humans and of the systems with which the drugs interact. Almost all types of variables used to assess the activity of drugs in the human CNS have already been applied in pharmacokinetic/pharmacodynamic research. However, compared with more traditional approaches to quantify the pharmacodynamics of neuropsychotropic drugs, the EEG method has the advantage of being objective, sensitive, continuous and reproducible. The present review focuses mostly on benzodiazepine pharmacology. A selection of some basic aspects that can be covered using pharmaco-EEG and pharmacokinetic/pharmacodynamic modeling from in vivo studies performed in humans in pharmacological research will be introduced: i) determination of the pharmacological characteristics of a compound; ii) comparison of potencies among drugs; iii) comparison of efficacy among drugs; iv) tolerance development; v) metabolite role; vi) enantiomers; vii) drug-drug interactions; viii) circadian rhythms; ix) factors affecting the observed effect; and x) the gain of physiopathological information about the systems with which drugs interact. Looking at the quantity and quality of the results obtained for the benzodiazepines, pharmacokinetic/pharmacodynamic modeling using EEG measures appears to be an ideal tool, and is potentially useful for other drugs acting on the CNS. PMID- 12575481 TI - Uses of pharmaco-EEG and pharmacokinetic-pharmacodynamic modeling in the clinical scenario. AB - In order to place pharmaco-EEG within the clinical context, the distinction between biomarkers, surrogate endpoints, clinical endpoints and clinical outcomes is introduced. State-of-the-art applications of pharmaco-EEG, together with pharmacokinetic-pharmacodynamic modeling in everyday clinical practice in anesthesiology (semilinear canonical correlation), psychiatry (discrimination between responders and nonresponders to pharmacological treatment using the test dose), neurology (antiepileptic field) and neurophysiology (first-order Markov model of sleep stage transitions) are discussed. The combination of both procedures, although successfully used during some drug development programs (opioids or benzodiazepines), is not widely applied in the clinical scenario where the central nervous system (CNS) is concerned. Much work is still need to develop fully the potentials that pharmaco-EEG together with pharmacokinetic pharmacodynamic modeling could bring to therapeutics in neuroscience. PMID- 12575482 TI - An introduction to statistical concepts for the analysis of EEG data and the planning of pharmaco-EEG trials. AB - The use of descriptive statistics and graphics for electroencephalogram (EEG) data after initial data reduction and the application of hypothesis tests to EEG data are described. In planning pharmaco-EEG trials, some notion of statistical power and variability of effect size is necessary as they are used to calculate sample size in parallel-group clinical trials. The need for sensitivity analysis during the study planning phase is demonstrated as well. Because EEG data are highly dimensional data, an outline of methods for reducing dimensionality is provided. These concepts are demonstrated using data from patients with Alzheimer's disease and from healthy controls. PMID- 12575483 TI - Recording EEG and EP data. AB - Recording evoked potentials (EPs), electroencephalogram (EEG) signals and polysomnographic biosignals requires particular attention with respect to amplification, filtering, and data handling. The present paper is aimed at providing a short overview of the fundamentals on recording techniques and the basics on biosignal processing methods used in EEG studies. It also examines the aspects associated with the reproducibility of EEG measurements, as well as the recording duration by Pharmaco-EEG (PEEG) studies. PMID- 12575484 TI - Planning of pharmacodynamic trials: I. Specificity and possible solutions. II. Stability considerations under placebo and interpretation of drug effects on EEG. AB - This workshop deals with the concept of quantitative electroencephalography (QEEG) to characterize the central effects of drugs. For proper interpretation, the circumstances under which data are obtained play an important role. To infer the size of variability in standard practice, we elaborated some computations for "placebo-treatment" in healthy volunteers, which helps to determine the threshold of drug effect detection. Simple rules for interpretation of multiple statistical comparisons were proposed and validation of dose effects were carried out with accepted reference compounds. Furthermore, psychotropic agents with comparable therapeutic indications often present similar modifications in EEG spectral composition (pharmaco-EEG profile). To extrapolate this concept, quantified wake EEG is a rapid, validated technique for early psychopharmacological investigation of new psychotropic compounds (Phase I, healthy volunteers). Classification of drug-induced changes in cerebral activity at this stage forms a useful decision instrument in planning the long-term clinical scenario of drug development (Phases II and higher). PMID- 12575485 TI - Combination of magneto- and electroencephalography in studies of monoamine modulation on attention. AB - The brain monoamines serotonin and dopamine have an important role in the regulation of human cognitive functions. Neural correlates of attention can be studied in millisecond resolution with magnetoencephalography (MEG) and electroencephalography (EEG), which provide complementary views on attentional processing. During "selective attention", a processing negativity (PN) overlaps EEG response to the attended tones. This component can be modulated with drugs affecting dopamine transmission in the brain, such as haloperidol and droperidol, whereas no effects seem to be caused by acute tryptophan depletion (ATD), decreasing serotonin synthesis in the brain. Distinct responses are associated with "involuntary attention". Responses reflecting sound-change detection and initiation of involuntary attention, the mismatch negativity and its magnetic counterpart, appear to be modulated by ATD, but not with haloperidol. Subsequent P3a elicited by actual attention shifting, as well as reorienting negativity reflecting orienting back to a relevant task, are in turn decreased by haloperidol, but not affected by ATD or other serotonin modulators. Serotonin may affect the earliest "preattentive" phases on auditory processing, indexed by mid latency magnetic responses and NIm, but haloperidol's effects on these parameters are insignificant. Taken together, serotonin and dopamine may have differential effects on attentional processing depending on time after stimulus presentation. PMID- 12575486 TI - Biomarkers and proof of concept. AB - Starting with an overview over the early human phase of drug development, the Pearson index, a normalized measure for the financial net present value of a drug development project, is introduced and it is shown how the value of obtaining critical information early can be quantified. Biomarkers used in this phase can be a means to obtain such information. Biomarkers can be classified as mechanistic, empirical or model based. Proof of concept trials are used to obtain the information. These trials can be proof of mechanism if they use a biomarker to investigate their mode of action, proof of viability if they investigate tolerability of putatively therapeutic doses or proof of efficacy if they use a clinical endpoint in a selected, homogenous population. If these trials are to decide on further development of a candidate drug, traditional statistical methods are not adequate. In the framework of decision analysis. Bayesian statistics offer an alternative. The basic techniques of this concept are introduced. The role of clinical electrophysiology is discussed in this framework. PMID- 12575487 TI - Stimulus- and event-related evoked potentials: from neuroscience to clinical neuropharmacology. AB - Synaptic neural (and neural system) functions are peculiarly sensitive to neuroactive compounds. Pharmacological interference/modulation is readily reflected by modifications in the organization of central nervous system (CNS), electrophysiologic signals occurring spontaneously in response to sensory stimulation (stimulus-related or evoked responses) or elicited in conjunction with sensory, motor or cognitive events (event-related potentials). Evoked responses reflect the basic physiology of sensory processes, while event-related potentials combine the time/space resolution of electrophysiologic signals with the specificity of eliciting neuropsychological conditions. The rationale for investigating drug effects on evoked and event-related potentials is manifold. Both are related to sensory and operant behavior and under suitable experimental conditions allow interpretation of drug-related changes in terms of CNS excitability. Some continuity between observations in man and in vivo or in vitro animal data is often possible. Proper handling of the stimulus physical properties or experimental/situational links may allow the responses to be related to sensory input or to neuropsychological manipulation of selectively activated CNS functions or functional subsystems and therefore to control spontaneous variability. This review summarizes today's knowledge of the application of electrophysiology to human neuropharmacology, with due reference to basic pharmacology and experimental evidence. PMID- 12575488 TI - Proof of safety of drugs: focus on vigilance. AB - It is well accepted that all new compounds, before administration to patients, should undergo safety evaluations in healthy subjects, including central nervous system (CNS) toxicity and as such the assessment of vigilance effects a relevant hallmark. The original concept of vigilance as a phenomenon observed only under conditions of monotony and signal regularity is increasingly falling into disfavor, embracing at present a much broader spectrum of behavior. Currently, vigilance may be regarded as a "readiness to adopt the appropriate behavior in a given situation, which thus finds outward expression through the quality and quantity of the behavior occurring in response to a given (internal or external) stimulus situation". The assumption that vigilance is a multifactorial phenomenon and not merely EEG data should be taken into account in order to study it accurately. Specifically, in drug research, apart from subjective reports and psychomotor performance tests, neurophysiological evaluations are regularly used such as Multiple Sleep Latency Test, Vigilance Epoch Classification or Parameters within a continuous scale. Although with limitations, temporal patterns of changes in activity of different frequency bands, indexes as the alpha slow-wave or the alpha anteriorization, computed from the EEG quantification, yielded different definitions of the intermediate states of the transition from wakefulness to sleep through the so-called subvigil stages. Spatial patterns are less documented. The recently proposed mathematical models to explain and predict variations in alertness are presented. Examples of the effects of different classes of drugs with the methods reported and its theoretical and practical relevance to vigilance research are introduced. PMID- 12575489 TI - The place of electrophysiology in the proof of concept approach of psychotropic drug development. PMID- 12575490 TI - Basics of PK-PD using QEEG: acute/repetitive administration, interactions. Focus on anxiolytics with different neurochemical mechanisms as examples. AB - Utilizing computer-assisted quantitative analysis of the electroencephalogram (EEG) in combination with certain statistical procedures and under specific design conditions, it is possible to objectively evaluate the functional bioavailability of psychotropic substances in the target organ: the human brain. Specifically, one may determine whether a drug is active in the central nervous system (CNS) compared with placebo in humans, the dose effect (including nonmonotonic drug effects along the continuum range of concentrations) and the time effect (including time-dependent pharmacodynamic phenomena as tolerance and sensitization), as well as its activity in relation to the formulation and route of application. Methodological aspects are introduced, discussing the usefulness of evaluating different treatments, doses, time points, states, target variables, electrodes and even different groups. Several issues are raised in relation to acute vs. repetitive administration, particularly those dealing with statistical comparisons when making conclusions about acute, repetitive or superimposed effects, and in relation to human psychotropic interactions, such as mechanistic drug-drug interaction descriptions, drug metabolites and enantiomers, as well as the importance of acquiring drug plasma concentrations, elapse of time and topographic distributions to accurately identify its occurrence. PK-PD modeling is introduced as a tool to enlarge the scope of inferences that can be derived when using pharmaco-EEG. The examples presented in order to develop the arguments are mainly focused on anxiolytic compounds belonging to the different neurochemical groups, benzodiazepines and azaspirones. Questions that have yet to been resolved are also addressed. PMID- 12575491 TI - Clinical applications of pharmaco-EEG in psychiatry: the prediction of response to treatment with antipsychotics. AB - The prediction of response to psychotropic drugs is of enormous interest in psychiatry, since absent or incomplete response to treatment may extend hospitalization time, increase direct and indirect costs of illness and, according to the so-called neurotoxicity hypothesis, worsen the illness prognosis. In the first part of the presentation, basic assumptions of pharmaco EEG relevant to the topic of the prediction of clinical response to antipsychotic treatment are summarized. The first step in the approach to this topic is the characterization of the quantitative EEG (QEEG) profile of the investigated drug. This is generally accomplished by assessing QEEG changes induced by a single dose of the same drug to groups of healthy subjects (test dose procedure). The prediction of patient responsiveness to a specific drug is based on the assessment of the similarity of their QEEG profile to the drug characteristic profile. In the second part of the presentation, main findings concerning QEEG changes induced by the acute administration of standard neuroleptics or novel antipsychotics in healthy subjects and in patients with schizophrenia are described. In the conclusion, achievements, limitations and future perspectives of the research field are outlined. PMID- 12575492 TI - Functional imaging with low-resolution brain electromagnetic tomography (LORETA): a review. AB - This paper reviews several recent publications that have successfully used the functional brain imaging method known as LORETA. Emphasis is placed on the electrophysiological and neuroanatomical basis of the method, on the localization properties of the method, and on the validation of the method in real experimental human data. Papers that criticize LORETA are briefly discussed. LORETA publications in the 1994-1997 period based localization inference on images of raw electric neuronal activity. In 1998, a series of papers appeared that based localization inference on the statistical parametric mapping methodology applied to high-time resolution LORETA images. Starting in 1999, quantitative neuroanatomy was added to the methodology, based on the digitized Talairach atlas provided by the Brain Imaging Centre, Montreal Neurological Institute. The combination of these methodological developments has placed LORETA at a level that compares favorably to the more classical functional imaging methods, such as PET and fMRI. PMID- 12575493 TI - Classification and evaluation of the pharmacodynamics of psychotropic drugs by single-lead pharmaco-EEG, EEG mapping and tomography (LORETA). AB - Utilizing computer-assisted quantitative analyses of human scalp-recorded electroencephalogram (EEG) in combination with certain statistical procedures (quantitative pharmaco-EEG) and mapping techniques (pharmaco-EEG mapping), it is possible to classify psychotropic substances and objectively evaluate their bioavailability at the target organ: the human brain. Specifically, one may determine at an early stage of drug development whether a drug is effective on the central nervous system (CNS) compared with placebo, what its clinical efficacy will be like, at which dosage it acts, when it acts and the equipotent dosages of different galenic formulations. Pharmaco-EEG profiles and maps of neuroleptics, antidepressants, tranquilizers, hypnotics, psychostimulants and nootropics/cognition-enhancing drugs will be described in this paper. Methodological problems, as well as the relationships between acute and chronic drug effects, alterations in normal subjects and patients, CNS effects, therapeutic efficacy and pharmacokinetic and pharmacodynamic data will be discussed. In recent times, imaging of drug effects on the regional brain electrical activity of healthy subjects by means of EEG tomography such as low resolution electromagnetic tomography (LORETA) has been used for identifying brain areas predominantly involved in psychopharmacological action. This will be demonstrated for the representative drugs of the four main psychopharmacological classes, such as 3 mg haloperidol for neuroleptics, 20 mg citalopram for antidepressants, 2 mg lorazepam for tranquilizers and 20 mg methylphenidate for psychostimulants. LORETA demonstrates that these psychopharmacological classes affect brain structures differently. PMID- 12575494 TI - Perceptual and cognitive event-related potentials in neuropsychopharmacology: methodological aspects and clinical applications (pharmaco-ERP topography and tomography). AB - Middle latency and late components of event-related brain potential (ERPs) are closely related to perceptual and cognitive information processing, respectively. In a double-blind, placebo-controlled study, the acute effects of lorazepam (2 mg), haloperidol (3 mg), methylphenidate (20 mg) and citalopram (20 mg) on ERP latencies, amplitudes, topographies and tomographies were investigated in 20 healthy subjects of 23-34 years of age. After automatic artifact minimization and rejection, standard N1 and P2 and target N2 and P300 components were determined. The tranquilizer lorazepam prolonged P300 latency, which indicates an impairment of stimulus evaluation time. Low-resolution brain electromagnetic tomography (LORETA) revealed decreases in N1 and P300 source strength in those brain regions with relevant generators of these components, which reflects impairments of attentional and cognitive processing resources. The neuroleptic haloperidol decreased N1 and P300 source strength predominantly in those brain regions not involved in the generation of these components, suggesting a shift of resources. The psychostimulant methylphenidate increased P300 source strength in brain regions with major P3b generators, indicating increases in energetic resources associated with stimulus encoding. The antidepressant citalopram increased N1 and P3b source strength in multiple brain regions specifically in the left prefrontal cortex, a brain region in which reduced blood flow and metabolism was found in depressed patients. PMID- 12575495 TI - Medicine on the Net Web Excellence Awards. PMID- 12575496 TI - Bad site design can turn users off. And good design helps them find what they're looking for. PMID- 12575497 TI - How to provide health material to consumers. PMID- 12575498 TI - Rising medical costs fuel employer interest in DM. PMID- 12575499 TI - Irritable bowel syndrome: practitioner awareness, patient education can limit utilization. AB - It may not be life-threatening, but IBS is more prevalent than either diabetes or asthma, and millions continue to access the health care system for relief because many clinicians aren't sure how to diagnose or treat the condition. In fact, there are a number of pharmacologic and non-pharmacologic interventions that can help sufferers better manage their condition while keeping the lid on utilization. See how clinicians can best recognize and do battle with this somewhat mysterious disorder. PMID- 12575501 TI - Perspectives. What we worry about when we worry about cost. PMID- 12575500 TI - Upsurge in weight loss surgery presents new opportunities for DM behavioral approach. AB - As many as 60,000 to 100,000 Americans are expected to undergo gastric bypass operations this year as a way to shed extra pounds that have turned into morbid obesity. However, experts suggest that many if not most of these individuals will require nutritional guidance and psychological support to help them cope with their drastically changed bodies in such a way that the benefits of surgery will be maximized over the long term. Unfortunately, not too many health care centers offer this kind of support. However, one therapist in Ambler, PA, is hoping to turn her DM-style behavioral program of psychological support into a model that others can follow. PMID- 12575502 TI - CMS clamps down on outlier payments. PMID- 12575503 TI - Using an intranet to reduce denials. PMID- 12575504 TI - Why I am optimistic. PMID- 12575505 TI - Association health plans high on Bush's '03 agenda. PMID- 12575506 TI - Compensation monitor. Managed care: an ever-growing part of physician income. PMID- 12575507 TI - Quality counts. So why not offer physicians bonuses? PMID- 12575508 TI - Changing medical evidence brings shift in C-section stance. PMID- 12575509 TI - Getting serious about generics. PMID- 12575510 TI - The quandary of compounding for MCOs: administrative costs, risks, and waste. AB - PURPOSE: Pharmacy compounding of prescription therapies that do not have Food and Drug Administration (FDA) approval increases administrative costs to managed care organizations and may also place members at risk for poor outcomes. This paper provides health care administrators with information and tools that can be used to manage risk-encouraged practices from pharmacy compounders, and suggests methods for evaluating the medical appropriateness and benefit of such compounds. STUDY DESIGN: Experiential findings. RESULTS: Methods indicate an effective way to manage this emerging pharmacy practice based on safety, risk avoidance, and outcomes. CONCLUSION: Focused management efforts can reduce the burden of reimbursement for compounding practices that are likely to present a greater risk of medical errors than commercially prepared medications. The use of existing terms in plan contracts, such as "experimental treatments or medications," "medical necessity," or "non-FDA approved agents," can enable an editing process that provides for appropriate benefit enforcement and control. PMID- 12575512 TI - Managed care outlook. AMA seeks closer monitoring of health plan mergers. PMID- 12575511 TI - Is outsourcing a cure for the IT blues? PMID- 12575513 TI - Perspectives. Health person of the year. PMID- 12575514 TI - A glimpse into the future. PMID- 12575515 TI - Keystone Health Plan West increases capitation rates. PMID- 12575516 TI - Risk-based capital guidelines give incentive to capitate. PMID- 12575517 TI - Provider groups, health plans use software to get accurate estimates of capitated expenses. AB - Information technology is improving the ability of health care organizations to predict annual expenses for groups of patients and to fairly allocate payments to providers based on health risk adjustments. Here's one capitated group's successful formula. PMID- 12575518 TI - IT products save small groups money under capitation. AB - Small physician groups now are able to pick and choose among affordable freestanding information technology (IT) products that can help practices reduce costs and improve quality. Read about a new report on IT solutions prepared by First Consulting Group for the California HealthCare Foundation. PMID- 12575519 TI - Use effective RM strategies to avoid errors and preserve capitation profits. AB - Risk management strategies employed by capitated groups range from purchasing reinsurance to paying physicians to take risk management continuing medical education courses. While few groups have in-house risk management programs beyond reinsurance, a growing number of medical groups and IPAs are practicing passive forms of risk management. PMID- 12575520 TI - The shape of European business in 2003. PMID- 12575521 TI - Public confidence in medical technology. AB - Three headlines have appeared in the British press recently, all of which concern health-care issues related to technology, and which have profound implications for public confidence in medical practice. Once again, at the year's end, it is necessary to reassess our own procedures and safeguards to prevent erosion of public confidence in our industry. PMID- 12575522 TI - Strategic drivers of contract manufacturing: Part I, The theory. AB - Medical device manufacturers and diagnostics companies have significantly increased their use of contract manufacturers to outsource production of components. This, the first of a two-part article, reviews strategic benefits and best practices in outsourcing. PMID- 12575523 TI - Small is the big issue: micromoulding. AB - Micromoulding techniques are allowing the industry to create a host of exciting new products. This article describes one micromoulding process and its benefits. PMID- 12575524 TI - The technological edge. PMID- 12575525 TI - Virtual medical devices. AB - Virtual medical devices are set to have a major impact on the industry. This article explains the practicalities and benefits of a new approach to designing and constructing many types of medical devices. PMID- 12575526 TI - FDA user fee and Modernisation Act. AB - A new law authorises the Food and Drug Administration to collect fees on medical device premarket submissions. The law also introduces important medical device regulatory reforms. This article discusses this new law and comments on the impact that it could have on the medical device industry. PMID- 12575528 TI - Foreign Establishment Registration and United States agent requirements. AB - In 2002 the Food and Drug Administration implemented new requirements for foreign companies that import medical devices into the United States (US). This article reviews the Establishment Registration requirements and discusses the new Establishment Registration and US Agent requirements for non-US companies. PMID- 12575527 TI - Standardising. New medical technologies, Part II. PMID- 12575529 TI - Medical devices on trial, Part I. AB - This is the first of a two-part article exploring the place that clinical research plays in today's medical device industry. Part I looks at the differences in development processes between drugs and devices and examines future trends for increased requirements for clinical data in the device sector. PMID- 12575530 TI - The UK market today and tomorrow. PMID- 12575531 TI - The future belongs to Ireland. PMID- 12575532 TI - [Telepathology: current problems and perspectives on development]. AB - Telepathology is a rapidly developing discipline of XXI-century medical technology. Advantages, shortcomings and perspectives of this method are discussed. PMID- 12575533 TI - [Adrenocortical tumors with neuroendocrine differentiation]. AB - 18 adrenocortical tumours and 5 pheochromocytomas were studied immunohistochemically. Expression of synaptophysin and chromogranin A was found in cells of cortical adenomas, "frontier" neoplasms and in 20-75% of carcinoma cells, this ultramicroscopically was confirmed by observation of typical neuroendocrine granules. Some groups of cells of cortico-medullary tumours also expressed proteins of neural differentiation (protein S-100). PMID- 12575534 TI - [Morphological, cytogenetic and molecular biological characteristics of lung cancer in persons exposed for a long time to radionuclide radiation pollution in the Semipalatinsk region of Kazakhstan]. AB - Surgical and biopsy material was used from 57 patients with lung carcinoma (LC): 17 patients (group I) lived close to testing area from the childhood to 1993 and were exposed to radiation at the year dose 0.1 ber. For comparison surgical and biopsy material from 40 patients (group 2) was taken who lived in the territories of Kazakhstan (10 patients) and Moscow (30 patients) with normal radiation background. The following features of LC were found in the groups in question: constant observation of dust particles and interstitial and focal fibrosis in tumours of various localization and histological type; prevailing small cell carcinoma as compared to other histological types; high frequency of nonsmall cell lung carcinoma; higher expression of IGFII, IGFBP1,2 and rare expression of IGFB3; proliferative activity followed by an increased expression of c-myc, bcl 2. PMID- 12575535 TI - [Cell composition of bronchoalveolar lavage fluid in compound chest trauma with main damage to the lungs]. AB - Lethal effect on alveolar macrophages is observed cytologically in all cases. Increase of neutrophils in alveols was found against the background of allergisation and inflammatory immune reactions. General state of patients with a syndrome of acute pulmonary damage but without parallel pulmonary pathology improved by day 5-20, neutrophillosis was moderate (22.8 +/- 8.3). On the contrary lavage liquid in patients with parallel pulmonary pathology had a high neutrophillosis (85.5 +/- 6.6). Endopulmonary cytogram allows prognosis and correction of treatment. PMID- 12575536 TI - [Diagnosis of chronic lymphoid leukemia (an immunomorphological study)]. AB - 31 cases of chronic lymphoid leukemia (CLL)/lymphoma from small lymphocytes (LSL) was studied immunomorphologically. All cases had B-phenotype and positive expression of markers CD 5, CD 23. Most characteristic for CLL/LSL was pseudofollicular character of tumour growth. Two morphological variants of CLL/LSL having similar phenotype were distinguished: typical variant with roundish conturs of tumour cells and atypical with centrocytoid morphology of tumour cell nuclei. PMID- 12575537 TI - [Expression of endometrial proteins in the uterine glands in physiological and non-developing pregnancy]. AB - It is established that at the 1st trimester of the progressing pregnancy uterine glands produce both endometrial proteins (placental al-microglobulin--PAMG) and alpha 2-fertility microglobulin (AFMB) but with different intensity: maximum level of PAMG is at early periods (4-5 weeks) but 8-9 weeks of gestation for AFMB. Production deficiency of PAMG and AFMB in the glands epithelium is found in 90% cases of non-developing pregnancy (4-7 weeks) and this is obviously one of the causes of early pregnancy interruption. PMID- 12575538 TI - [Pathogenetic basis of hormonal endometrial hypertrophy in early spontaneous abortions]. AB - Pathogenetic role of hormonal dissociation produced by disturbances of biochemical transformation of testosteron to estradiol in the mechanism of deficient gestational endometrial transformation is shown. PMID- 12575539 TI - [Morphological and functional characteristics of thyroid glands from dead fetuses and their hormonal status in gestational hyperthyroidism]. AB - A complex organometric, stereohistometric and immunomorphological investigation of thyroid gland of intranatally decreased mature fetuses from women with hyperthyroidesm is performed. Parallel study of thyroid hormones levels in a cord blood and thyroid status of woman in childbirth, on the one hand, and study of morphological peculiar properties and morphometric parameters of fetal thyroid gland, on the hand, allows us to obtain a clear idea on morphological insufficiency of their thyroid gland on the eve of beginning of their extrauterine life. It also throws some light on some pathogenic elements of disfunctions of thyroid status in the newborn of women with hyperthyroidism. PMID- 12575540 TI - [Transformed and pathological red cells in endogenous intoxication and extracorporeal detoxication]. AB - Peripheral red blood cells obtained from 227 urological patients with chronic renal failure and purulent intoxication (78 of them were treated by hemodialysis and hemosorption) are examined. Scanning electron microscopy, cell surface charge, lipid peroxidation in erythrocytes, blood viscosity data are summarized. Significant morphofunctional changes dependent on the severity of the intoxication and the detoxication efficiency are manifestation of nonspecific features of the cell injury and adaptation. Their complex and dynamics are suggested as important diagnostic criteria. PMID- 12575541 TI - [Ultrastructural changes of child rib cartilage in different deformations of chest cells]. AB - Different chest deformations are accompanied by appearance of asbestos-like fibrills in pericellular matrix, development of significant lipid and carbohydrate chondrocyte dystrophy, irreversible changes in their majority. Anomalous accumulation of intermediate filaments, lipid and glycogen, may substitute the cytoplasm organells in Marfan's syndrome. Pathologic disturbances of costal cartilage, connected with ultrastructural changes of chondrocytes, intercellular substance and extracellular matrix are morphologic substrate of different chest deformations. PMID- 12575542 TI - [Changes in immunogenesis organs after thyroidectomy and hormonal correction in an experiment]. AB - Regulating effect of thyroid hormones on functions of organs of the immune system has been demonstrated in the experiment. Deficiency of thyroid hormones affects thymulin synthesis and morphofunctional status of immunogenesis organs. Combination of thymostimulin and thyroxin in the thyroidectomized animals, as compared to their separate use, is pathogenetically the most adequate correcting therapy aiming normalization of structural and functional disorders in the thymus and spleen. Prolonged reaction of tissual basophils saturated with heparin, indicated hormonal activation of blood anticoagulative system, and complex of the above hormones can be used for prevention of thromboembolic complications. PMID- 12575543 TI - [Tumors of the thymus]. AB - Tumours of the thymus include heterogenic group of benign and malignant organ specific (thymomas and carcinoma) and organ-nonspecific (neuroendocrine, germinogenic tumours, lymphomas and others) neoplasms. Histologic classifications are now the most useful for the correlation with prognosis and clinical manifestations. It is obligatory to do verification of the progression stage. PMID- 12575545 TI - [To seek on the rough and bumpy road]. PMID- 12575544 TI - [Role of chronic viral hepatitis in the development of hepatocellular carcinoma]. AB - Role of HBV- and HCV-infections, liver cirrhosis, alcohol in the development of hepatocellular carcinoma, and hepatocyte dysplasia is considered. PMID- 12575546 TI - [Comparative study on efficacy of qinggan granule and bushen granule in treating chronic hepatitis C]. AB - OBJECTIVE: To evaluate the efficacy of Qinggan Granule (QGG) and Bushen Granule (BSG) in treating chronic hepatitis C (CHC) from blood donors clinically, biochemically and pathologically as well as to explore the therapeutical principle and methods of TCM. METHODS: Sixty-six patients with CHC were divided into three groups, the 36 patients in the QGG group treated with QGG, the 18 patients in the BSG group treated with BSG and the 12 patients in the control group untreated. The dose of QGG and BSG given was 30 g each time, three times per day for 6 consecutive months. Clinical and serum biochemical parameters as well as pathological change of liver biopsy before and after treatment were observed dynamically and compared. RESULTS: After treatment, in the two treated group, clinical symptoms were improved significantly, alanine transaminase (ALT) and aspartate aminotransferase (AST) reduced markedly. Moreover, QGG showed the effects of increasing albumin and lowering alkaline phosphatase (ALP). On the contrary, ALT still remained as before but AST further increased in the control group. Pathological examination showed that the inflammatory grade (IG) decreased in 42.2% and fibrotic stage (FS) decreased 21.2% of the patients in the QGG group, but no significant changes of the two indexes occurred in the BSG group, while in the control group, IG unchanged and FS increased in 3 cases. Chevallier's semi-quantity system analysis showed significant decrease of both scores in the QGG group (P < 0.05), only decrease of IG in the BSG group, and slight raise of IG and significant increase of FS (P < 0.05) in the control group. CONCLUSION: Condition of chronic hepatitis C patient would deteriorate progressively if not treated in time, especially the development of fibrosis. QGG and BSG could improve the clinical symptoms significantly, lower ALT and AST, eliminate inflammatory damage in the liver, slow down or reverse liver fibrosis process with stable long-term effect. Though most of the patients present a Syndrome of Liver-Kidney Yin-deficiency, the effect of QGG is superior to that of BSG, suggesting that the treatment of CHC should mainly be clearing principle, and method of clearing Liver and eliminating Dampness may be more suitable for them. PMID- 12575547 TI - [Clinical and experimental study of shuangcao tuihuang granule-1 in treating severe jaundice of acute icterohepatitis]. AB - OBJECTIVE: To observe the effect of Shuangcao Tuihuang Granule-1 (SCTH-1) in treating severe jaundice of acute icterohepatitis and to study its mechanism. METHODS: Thirty-four patients with severe jaundice of acute icterohepatitis were treated with SCTH-1, their therapeutic effects were analyzed. In the experimental studies, the influences of SCTH-1 on acute liver injury, liver superoxide dismutase (SOD) and serum ALT and AST were monitored. RESULTS: The markedly effective rate and effective rate of the treated group were 73.5% and 23.5% respectively. The markedly effective rate of the treated group was obviously higher than that of the control group (P < 0.05). Experimental study revealed that SCTH-1 could reduce the level of serum transaminase and inflammation of liver tissue in mice with acute liver damage. In addition, SCTH-1 could raise the activity of liver SOD (P < 0.05). CONCLUSION: SCTH-1 could accelerate the jaundice subsidence, improve the liver function and ameliorate the liver injury, its mechanism was possibly correlated with raising SOD activity, scavenging the oxygen free radicals. PMID- 12575548 TI - [Relationship between liver fibrosis criteria and syndrome-type of TCM in patients with non-alcoholic fatty liver]. AB - OBJECTIVE: To explore the relationship between the liver fibrosis criteria and TCM Syndrome, type in the patients suffering from non-alcoholic fatty liver. METHODS: Seventy-nine patients with non-alcoholic fatty liver diagnosed by B ultrasonography were classified by TCM Syndrome Differentiation and their liver fibrosis criteria was determined and compared with those of the health subjects as control. RESULTS: Levels of procollagen III (PCIII), hyaluronic acid (HA), collagen IV (CIV) and laminin (LN) in the fatty liver group were significantly higher than those in the control group (P < 0.05, P < 0.01). In respect to the TCM Syndrome-types, PCIII, CIV and LN in patients of Phlegm-stasis combined type were significantly higher than those in patients of other two Syndrome-types (P < 0.05, P < 0.01), while HA was insignificantly different (P > 0.05). CONCLUSION: There was trend of liver fibrosis in fatty liver patients. It was indicated that the Phlegm-stasis combined Syndrome-type possibly was the main TCM pathologic factor of the increasing of fibrosis criteria for non-alcoholic fatty liver. PMID- 12575549 TI - [Relationship between coronary angiography and syndrome differentiation type of TCM]. AB - OBJECTIVE: To understand the relationship between the coronary Angiography (CA) reflected degree of coronary artery lesion and the TCM Syndrome Differentiation type (TCM-SDT). METHODS: Patients TCM-SDT were differentiated adopting the standard of Syndrome Differentiation for coronary artery diseases revised in 1990, and the comparison of TCM-SDT with the findings in CA was carried out. RESULTS: CA examination showed that 29 patients (39.73%) were of one-artery lesion, 9 patients (12.33%) two-artery lesion and 35 patients (47.95%) three artery lesion; the stenosis degree of them were: 54 arteries were mild stenosis, 38 moderate stenosis and 31 severe stenosis; 29 arteries were completely occluded, the sum total of affected artery was 152 (69.41%), and the other 67 arteries (30.59%) had no lesion. TCM-SDT showed that all the 73 patients had secondary Excess Syndrome, among them 73 patients (100%) with blood stasis Syndrome, 39 patients (53.4%) with Phlegm-Turbid Syndrome, 16 (21.0%) with Qi stagnation Syndrome and 7 (9.6%) with Cold condensation Syndrome. 63 patients (86.3%) with obvious primary Deficiency Syndrome, among them, 57 (78.1%) were Qi Deficiency, 18 (24.7%) Yang-Deficiency, 28 (38.4%) Yin-Deficiency and 5 (6.8%) Yang collapse. All patients had blood stasis of various degrees, manifesting on tongue proper prominently. In the 80.8% patients who had angina pectoris, 74.0% had their mouth, lips and gum dark red, dark purple or light dark in color. The comparison between TCM-SDT and Blood-Stasis score revealed that patients of Cold condensation-blood stasis Syndrome and Yang collapse-blood stasis Syndrome were serious. Patients of Qi stagnation-blood stasis Syndrome and Yin Deficiency-blood stasis Syndrome were mild cases, while patients of Phlegm Turbid-blood stasis Syndrome and Qi Deficiency-blood stasis Syndrome were moderate severity cases. The number of affected artery in Cold condensation patients was the most, and in Yin-Deficiency patients was the least. The coronary artery stenosis degree in patients of Qi-stagnation and Yin-Deficiency were milder than in those of Yang collapse and Yang-Deficiency patients. CONCLUSION: The more the number of arterial lesion, the severer the degree of stenosis and the higher the blood stasis score, then the more serious the degree of stasis. PMID- 12575550 TI - [Preliminary study on relationship between TCM syndrome-type and insulin resistance in coronary heart disease]. AB - OBJECTIVE: To observe the relationship between TCM Syndrome-type and insulin resistance (ISR) in coronary heart disease (CHD). METHODS: Fifty patients were divided into 3 groups according to the Syndrome Differentiation-typing in TCM, the Heart blood stasis (HBS) Syndrome group, the Phlegm-Turbid stagnation (PTS) Syndrome group and both Qi-Yin Deficiency (QYD) Syndrome group. The fasting blood glucose (FBG), fasting blood insulin (Ins), insulin antibody (IAA), islet cell antibody (ICA), glutamic acid decarboxylase antibody (GAD-Ab) and related blood lipid parameters in patients were determined and insulin sensitive index (ISI) was calculated simultaneously. Then the above-mentioned data were compared with those determined in 20 healthy control subjects. RESULTS: The levels of FBG and Ins in CHD group were higher than those in healthy control group significantly (P < 0.05), but ISI level was obviously lower (P < 0.01). Moreover, the positive ratio of IAA (40%) was higher in CHD group than that in the control group (5%) significantly (P < 0.01). Comparison between the 3 TCM Syndrome-type groups and the control group showed that ISI level in HBS and PTS group was obviously lower than that in the control and the QYD (P < 0.05) respectively, and the IAA positive ratio in the former 2 groups (50%, 47.3%) was higher than that in the later two (5%, P < 0.01 and 15.38%, P < 0.05) markedly. While Ins level increased only in the HBS group (P < 0.05). Besides, patients of HBS and PTS were accompanied by lipid metabolic disturbance. CONCLUSION: ISR presents in part of CHD patients particularly in those with HBS and PTS, which was partly due to the existence of IAA in patients serum. PMID- 12575551 TI - [Effect of songling xuemaikang capsule combined with captopril on quality of life in primary hypertension patients]. AB - OBJECTIVE: To compare the hypotensive effects of captopril and combined medication of Songling Xue-maikang Capsule (SXC) with captopril on quality of life (QOL) in treating primary hypertension patients. METHODS: For 166 hypertension patients, a perspective randomized double blind study was performed with both sexes, aged 42-78 years, blood pressure (BP) ranged > or = 140/90 mmHg and < or = 200/110 mmHg. Patients were randomly divided into two groups: captopril group and combined medication group. Each patient should answer the QOL questionnaire independently before and after experiment. 60 age-matched normotensive subjects without chronic diseases served as control. RESULTS: Both hypertension groups showed an improvement of BP and QOL after treatment, but the combined medication group had a higher score in sense of well being, physical symptom-signs, work performance and life satisfaction than the captopril group had. CONCLUSION: Combined medication of captopril and SXC had better effect than captopril alone either in lowering BP or in improving QOL of primary hypertension patients. PMID- 12575552 TI - [CEA comprehensive evaluation for Western and traditional Chinese hypotensive drugs]. AB - OBJECTIVE: To make a cost-effectiveness analysis (CEA) comprehensive evaluation for 4 hypotensive drugs, by observing the effects of drugs on blood pressure lowering, symptom improving, and adverse effect and quality of life (QOL) influencing. METHODS: Two hundred and ninety-two patients with mild to moderate hypertension were divided into 4 groups at random and treated with compound Lingjiao Jiangya Pill, benazepril, amlodipine and indapamide respectively with a therapeutic course of 6 weeks. QOL was measured with FS-36 questionnaire and efficacy policy model was applied for comprehensive evaluation of CEA. RESULTS: Through CEA a comprehensive evaluation was made in order as follows: indapamide 3.65 which was the best, and then Lingjiao Jiangya Pill 3.55, amlodipine 2.90 and benazepril 2.35. CONCLUSION: The 4 drugs can not only lower blood pressure but also improve clinical symptoms with less adverse effect. It is of great practical significance to evaluate hypotensive drugs by combining QOL appraisal. The position of Lingjiao Jiangya Pill in CEA order suggests that more attention should be paid to herbal hypotensive drugs. PMID- 12575553 TI - [Effect of yiqi bushen huoxue herbs in treating children asthma and on levels of nitric oxide, endothelin-1 and serum endothelial cells]. AB - OBJECTIVE: To observe the therapeutic effect of Yiqi Bushen Huoxue herbs (YQBSHX, the Chinese herbs for supplementing Qi, replenishing Kidney and activating blood circulation) on children asthma in regard to its effect on the serum levels of nitric oxide (NO), endothelin-1 (ET-1) and circulating endothelial cells (CEC). METHODS: Two hundred children with asthma were divided into two groups, the YQBSHX and the control group, and their serum levels of NO, ET-1 and CEC were detected in acute and remission stage respectively. Twenty cases in each group were followed-up for 1 year to observe the frequency of asthma attack and the changes in ET-1, NO and CEC levels. RESULTS: Serum levels of ET-1, NO and CEC in patients of acute stage were obviously raised. After treatment, the above mentioned criteria in the YQBSHX group were significantly lower than those in the control group (P < 0.01). And the frequency of attack in the YQBSHX group was markedly reduced. CONCLUSION: YQBSHX herbs could reduce ET-1, NO and CEC levels in children asthma, lessen the frequency of attack, therefore, to elevate the cure rate of children asthma. PMID- 12575554 TI - [Quantitative study on tongue diagnosis in stroke patients]. AB - OBJECTIVE: To study the tongue proper and tongue fur quantitatively in stroke patients. METHODS: Three hundred and seventy-eight stroke patients were studied by means of TCM Glossoscopy Expert System, the techniques of color digitized image processing and pattern recognition were applied to objectively study the changes of tongue proper and tongue fur. RESULTS: Dark-red tongue was the commonest tongue proper, which revealed in 128 cases (33.86%) in the 378 patients investigated. There was significant difference of red (R), green (G) and blue (B) values in dark-red tongue as compared with those in pale-purple tongue, pale white tongue, pale-red tongue, deep red tongue, purple-red tongue and blue-purple tongue (P < 0.05, P < 0.01). Thick and yellow greasy fur was the commonest in 214 cases of acute stage, it accounted for 30.84% and with a total of 66 cases. There was also significant difference in R/G and R/B values among different types of tongue fur (P < 0.01). CONCLUSION: This quantitative study on stroke patients regarding to tongue proper and tongue fur by means of TCM Glossoscopy Expert System provides a base for standardization and quantification of diagnosis of stroke patients. PMID- 12575556 TI - [Study of Chinese herbal medicine in treating ascites and their mechanism in regulating lymphatic stomata]. AB - OBJECTIVE: To study the therapeutic effect of Chinese herbal medicines (CHM) in treating ascites to elucidate its mechanism in regulating the lymphatic stomata and promoting the absorption of ascites from the peritoneal cavity. METHODS: Using scanning electron microscope (SEM) and computerized image processing and quantitative analysis assays, the CHM extract consisting of Atractylodes macrocephala, Salvia miltiorrhiza, Codonopsis pilosula, Alismatis orientale and Leonurus heterophyllus were studied. RESULTS: Intraperitoneal injection of nitric oxide (NO) supplier or CHM administration could cause the average area of lymphatic stomata obviously enlarged (P < 0.05), and the open numbers significantly increased (P < 0.01) in normal healthy mice. When L-notroarginine, a NO synthetase suppressor, was injected after CHM administration, it was found that the regulating effect of CHM on lymphatic stomata was inverted obviously, i.e. the average area and the density of lymphatic stomata were markedly reduced (P < 0.01). CONCLUSION: CHM might treat ascites through increasing the endogenous NO concentration to open the lymphatic stomata and in turn to conduct the peritoneal water through lymphatic path. PMID- 12575555 TI - [Effect of tiaoxin recipe on beta-amyloid induced neurotoxic model of the primary cultured rat hippocampal neurons]. AB - OBJECTIVE: To study the effect of Tiaoxin Recipe (TXR) on beta-amyloid induced neurotoxicity in the primary cultured hippocampal neurons and the therapeutical mechanism of it. METHODS: In order to establish neurotoxic model, the primary cultured rat hippocampal neurons were treated with 25 mumol/L aggregated beta amyloid fragment 1-42 (A beta 1-42). Neuronal survival was assessed by lactate dehydrogenase (LDH) activity. Apoptosis of neurons was determined with transmission electron microscopy (TEM) examination and flowcytometric analysis. The protein expression of apoptosis-associated gene (Bcl-2, Bax, c-Jun) were examined by using immunocytochemical SABC method. RESULTS: In the A beta 1-42 treated neurons, the release of LDH was increased, TEM examination revealed that the cell body became shrunken, the chromatin was compacted or the patches of condensed chromatin lay against the nuclear membrane and the smooth surfaced endoplasmic reticulum was expanded. Flowcytometric analysis displayed the peak of hypodiploid DNA content. The expression of Bcl-2 was lowered, while Bax and c-Jun protein were increased. After treated with TXR, the above-mentioned changes were improved with the exception of c-Jun expression. In the Tacrine treated group, only the neuronal survival was increased, other observed criteria had insignificant change. CONCLUSION: TXR could attenuate the neurotoxic action of A beta 1-42 and improve neuronal survival via suppressing apoptotic process. TXR could inhibit the apoptotic process via regulating the expression of bcl-2 and bax. Tacrine could improve neuronal survival, but its inhibition on neuronal apoptosis was not significant. PMID- 12575557 TI - [Effect of dingxin recipe on intracellular free calcium, membrane potential and mitochondria membrane potential in normal and hypoxia myocytes of rats]. AB - OBJECTIVE: To elucidate the molecular mechanism on anti-arrhythmic function of Dingxin Recipe (DXR). METHODS: Cardial myocytes isolated by trypsin method were cultured and labelled with various fluorescence stains. DXR contained serum induced changes of cytolic calcium concentration [Ca2+]i, cell membrane potential (MP) and mitochondria membrane potential (MMP) were determined with laser confocal microscopy. RESULTS: Hypoxia caused increase of [Ca2+]i and MMP, and decrease of MP, while DXR contained serum lowered the [Ca2+]i in normal or hypoxian myocytes, improved hypoxia induced MP reduction and maintain MMP at baseline level under hypoxia circumstances. CONCLUSION: Through inhibiting the hypoxia caused [Ca2+]i and MMP elevation and MP lowering, DXR displays its action in protecting myocardial cells to prevent and treat arrhythmia. PMID- 12575558 TI - [Experimental study on xiaoliu pingyi mixture with medicated serum in inducing apoptosis of human hepatocellular carcinoma cell line H-7402]. AB - OBJECTIVE: To observe the efficacy of Xiaoliu Pingyi Mixture (XLPY) in inducing the apoptosis of human hepatocellular carcinoma (HCC) cell line H-7402. METHODS: Using serum pharmacologic method, the morphological change of apoptotic cells was observed at different time points by co-culturing with different concentrations of medicated serum containing XLPY with light and fluorescent microscopy, flow cytometry, and immunohistochemical staining methods. Blank serum was taken as control. RESULTS: The different concentration of medicated serum containing XLPY could significantly inhibit the growth of H-7402 cell line in vitro. After treated with medicated serum of XLPY, the cellular morphologic changes were typical apoptosis under light and fluorescent microscopy, the sub-G1 peaks were observed by flow cytometric assays. The apoptosis rate revealed a medicated serum concentration and time dependent manner. The control group 5%, 10% medicated serum induced apoptotic rate were 0.49%, 5.58% and 8.46% by 24 h; while by 48 h, it was 0.83%, 11.58% and 14.25% respectively. bcl-2 gene examination showed that the bcl-2 gene expression was inhibited. CONCLUSION: XLPY could induce apoptosis in carcinoma cells, and play the clinical role of anti-tumor and anti-metastasis. PMID- 12575559 TI - [Experimental study of reinforcing effect of bushen yijing capsule on bone quality of cancellous bone in ovariectomy caused rats osteoporosis model]. AB - OBJECTIVE: To explore the influence of Bushen Yijing Capsule (BSYJC) on bone quality of cancellous bone in ovariectomize induced osteoporosis rats model. METHODS: Forty Wistar female rats, 10 months old, were randomly divided into 4 groups, the normal group, the model control group, the premarin treated and the BSYJC treated group, 10 in each group. The normal group was only sham operated and the other three were ovariectomy performed and treated starting at day 91 postoperationally for consecutively 90 days. Rats were sacrificed at the end of experiment, the second lumbar vertebra was taken out to determine bone density and contents of calcium, phosphorus and organic substances, and the third lumbar vertebra taken out to measure the bone density and biomechanics properties. Bone quality was evaluated with the above indices. RESULTS: BSYJC could significantly raise the bone density, increase the contents of calcium, phosphorus and organic substances, and improve the biomechanical properties of the lumbovertebral bone in osteoporosis rats. CONCLUSION: BSYJC could improve the bone quality of the cancellous bone in ovariectomized rats. PMID- 12575560 TI - [Experimental study on effect of bushen huoxue xiezhuo decoction in treating minimal change nephropathy]. AB - OBJECTIVE: To explore the therapeutic effect and mechanism of Bushen Huoxue Xiezhuo Decoction (BHXD) in treating minimal change nephropathy (MCN) in rats. METHODS: The MCN model rats established by a single intravenous injection of Adriamycin were divided into the model, BHXD group, and a normal group was set up for control. The effect of treatment on renal function, hemorrheologic parameters, renal tissue TGF-beta 1 expression and polyanion sites on glomerular basement membrane were observed dynamically. RESULTS: After treatment, all the parameters between the BHXD group and the model group were significantly different respectively, morphological observation also showed the pathological changes in the BHXD group were milder than those in the model group. CONCLUSION: BHXD treatment could markedly improve the renal function, alleviate blood hypercoagulability and hyperviscosity, protect the anion barrier and delay the progression of glomerular fibrosis and sclerosis. PMID- 12575561 TI - [Experimental study on effect of zenggu-I, -II, -III on osteoblast in vitro]. AB - OBJECTIVE: To investigate the effect of Zenggu (ZG)-I, -II and -III on osteoblast (OB) cultured in vitro. METHODS: OB was isolated from calvaria of 22-day-old fetal SD rats by means of modified sequential collagenase digestion and incubated in RM1640 medium and the cell morphology was observed by alkaline phosphatase (ALP) staining. Different concentrations of ZG-I, -II and -III were added to the OB and incubated. The effect of ZG on the proliferation and osteogenesis of OB was monitored by MTT analysis, BGP content radioimmunoassay, ALP activity and calcium contents determination. RESULTS: The BGP, ALP, and calcium contents of the cultured OB cells as well as the cell proliferation were higher in ZG-III group than those in the other groups (P < 0.05) in a dose-dependent manner. It suggested that ZG-III could promote the proliferation and osteogenesis of OB, but ZG-I and ZG-II had insignificant influence on OB cultured in vitro. CONCLUSION: ZG-III could promote the OB proliferation and bone formation, ZG-I, -II, -III sequential therapy could effectively prevent and treat osteoporosis and promote the bone formation. PMID- 12575562 TI - [Discussion on the mutual academic supplementing of integrated traditional Chinese and Western medicine]. PMID- 12575563 TI - [Intervention effect of Chinese herbal medicine on changes of immune system in senile dementia]. PMID- 12575564 TI - [The sponsoring process of Chinese Association of the Integration of Traditional and Western Medicine]. PMID- 12575565 TI - [Clinical observation on effect of Xianlong capsule in treating vascular dementia]. AB - OBJECTIVE: To study the effect of Xianlong Capsule (XLC) in treating vascular dementia (VD) and to explore its therapeutic mechanism. METHODS: The criteria of learning and memory related cholinergic nerve system, oxygen free radical and nitrogen monoxide (NO) in 35 patients with VD treated with XLC and 33 patients treated with hydergine were observed and compared with the normal control group. RESULTS: XLC could lower the acetylcholine esterase activity, increase the acetylcholine content in erythrocytes and plasma and decrease the NO content. No toxic adverse reaction was found in the observation period. CONCLUSION: XLC has definite effect in treating VD in aged patients, the therapeutic mechanism may be related with the actions in strengthening cholinergic nerve system, improving oxygen free radical metabolism, regulating NO and relieving its excitatory neurotoxicity. PMID- 12575566 TI - [Clinical observation on effect of baisuifang oral liquid in treating vascular dementia]. AB - OBJECTIVE: To study the effect of Baisuifang Oral Liquid (BSF) in treating vascular dementia. METHODS: Patients selected according to the diagnostic standard of American Association of Psychiatry were randomly divided into two groups, the 37 patients in the BSF treated group and the 28 patients in the control group treated by Naofukang with therapeutic course of 3 months. Changes of clinical symptoms, TCM Syndrome, mini-mental state examination (MMSE) and activity of daily living (ADL) score, hemorrheologic characters, platelet aggregation and blood lipid were observed. RESULTS: BSF could ameliorate the symptoms and TCM Syndrome, decrease blood lipid, improve the hemorrheologic character, elevate the MMSE and ADL score of patients. The total effective rate in the treated group was 75.68% and the markedly effective rate 24.32%, which were obviously superior to those in the control group respectively. CONCLUSION: BSF is an effective Chinese herbal preparation in treating vascular dementia, which could promote the restoration of learning memory, and improve the clinical symptoms. PMID- 12575567 TI - [Clinical observation of effect on tongmai yizhi capsule in treating vascular dementia]. AB - OBJECTIVE: To observe the effect of Tongmai Yizhi capsule (TMYZC) on learning in patients of vascular dementia (VD). METHODS: Thirty patients of VD were treated by oral administration of TMYZC, the changes in them before and after treatment were observed and compared with those in the 20 patients treated with hydergine as control. RESULTS: The total effective rate in the TMYZC treated group was 70.67% while that in the control group was 70.00%, the difference between the two groups was insignificant. CONCLUSION: Both TMYZC and hydergine have the effects of dilating blood vessels, increasing the cerebral blood flow, lowering the hyper coagulative status and improving the free radical scavenging capability of patients. PMID- 12575568 TI - [Clinical study on long-term treatment of ankylosing spondylitis with integrated Traditional Chinese and Western medicine]. AB - OBJECTIVE: To seek for the new method of integrated traditional Chinese and western medical treatment (TCM-WM) for ankylosing spondylitis (AS) to control the disease development, shorten the therapeutic course and reduce disability rate. METHODS: Patients of AS enrolled in this study were 106 patients in group A, who were persistently treated with TCM-WM for over 10 months, 48 patients in group B treated with western medicine and 34 in group C treated with non-steroidal anti inflammatory and analgesic agents. They were followed-up for 2 years. RESULTS: (1) Short-term effect: the markedly effective rate in the three groups was 73.58%, 47.92% and 5.88% respectively, and the total effective rate was 96.23%, 87.50% and 41.18% respectively. The comparisons of group A to group B and C showed significant difference, the effect in group A was superior to that in group B and group C (chi 2 = 10.58, P < 0.01 and chi 2 = 67.76, P < 0.01 respectively). (2) Results of 2-year follow-up: the recurrence rate in group A was 18.37% (9/49), it was insignificantly different from that in group B (7/23 = 30.43%, chi 2 = 1.32, P > 0.05), but significantly different from that in group C (8/12 = 66.67%, chi 2 = 11.19, P < 0.01). CONCLUSION: Long-term TCM-WM therapy, which concentrated on the advantages of traditional Chinese and western medicine, was a practical method in treating AS with reliable effect, few side-reaction, therefore, is worth spreading. PMID- 12575569 TI - [Clinical observation on treatment of non-small cell lung cancer with Chinese herbal medicine combined with bronchial arterial infusion chemotherapy]. AB - OBJECTIVE: To explore the effect of treatment for non-small cell lung cancer (NSCLC) with Chinese herbal medicine (CHM) combined with bronchial arterial infusion chemotherapy (BAIC). METHODS: Patients with moderate or advanced NSCLC were randomly divided into two groups, the 39 patients in the treated group treated with CHM plus BAIC therapy and 37 in the control group treated with BAIC alone. The short-term therapeutic effect, long-term survival rate, changes of clinical principal symptoms, quality of life and peripheral blood pictures in the two groups were observed and compared. RESULTS: After treatment the rate of CR + PR + NC in the treated and the control group was 92.31% and 70.27% respectively, the inter-group comparison showed a significant difference (P < 0.05). The 0.5-, 1- and 2-year survival rate in the treated group was 79.49%, 56.41% and 51.28% respectively and in the control group, 72.97%, 51.35% and 24.32% respectively. The 2-year survival rate in the former was better than that in the latter (P < 0.05). Moreover, the improvement of clinical symptoms, Karnofsky scoring, body weight and peripheral blood picture in the treated group was superior to that in the control group. CONCLUSION: Therapeutic effect of BAIC could be enhanced by combining it with CHM. PMID- 12575570 TI - [Clinical observation on multiple organ dysfunction syndrome due to fish gall bladder poisoning]. AB - OBJECTIVE: To study the functional changes of liver, kidney, myocardium and gastro-intestine after fish gall bladder poisoning and the pathogenic mechanism of acute renal failure. METHODS: The liver and kidney function, myocardial enzyme, urinary N-acetyl-beta-D-glucosaminidase (N-AG), 24 hrs intake and output volume of liquid and kidney B-ultrasonographic examination in 11 patients of severe acute fish gall bladder poisoning were observed. And kidney biopsy was carried out in one patient under B-ultrasonography for understanding the renal pathological changes. RESULTS: All the 11 patients were cured with disappearance of clinical symptoms. After the poisoning the order of the severity of organ damage were kidney, liver, myocardium and gastro-intestine tract. The levels of blood creatinine, blood urea nitrogen, urinary N-AG, glutamic-pyruvic transaminase, total bilirubin were lowered significantly after treatment. Biopsy examination under light microscope showed toxic damage of renal tubules mainly in proximal tubules, and under electron microscope, the pathologic changes were mitochondrial vacuolar degeneration of tubular epithelial cells, swelling of epithelial cells, partial fusion of processes in glomeruli and narrowing of saccular cavity. CONCLUSION: Fish gall bladder poisoning could cause acute multiple organ dysfunction syndrome (MODS), the affected organs in order of severity of damage were kidney, liver, myocardium and gastro-intestine tract. In kidney the damage was mainly at the proximal tubules. Urinary NAG is a sensitive criterion for determining the diagnosis and prognosis of renal tubular dysfunction. PMID- 12575571 TI - [Immune functional changes in patients of acute Henoch-Schonlein purpura and regulatory effect of integrated Traditional Chinese and Western medicine on it]. AB - OBJECTIVE: To observe the changes of immune function in children with acute Henoch-Schonlein purpura (AHSP) and the effects of integrated traditional Chinese and western medicine (TCM-WM) on it. METHODS: Immunological criteria in TCM-WM group (n = 35) treated by Yinfu Decoction (YFD) combined with transfer factor and 35 patients in the control group (n = 35) treated by conventional treatment were observed and compared before and after treatment. Also the criteria were compared with those of 40 healthy subjects. RESULTS: Before treatment, the levels of IgA, IgM, CD4, CD4/CD8 ratio, rosette forming rate of RBC-C3b receptor in patients were higher than those in the healthy subjects, but the levels of CD8 was obviously lower, the difference was significant (P < 0.01). The above-mentioned criteria were all improved in the two treated groups after treatment, and the improvement was more significant in the TCM-WM group than that in the control group (P < 0.05). Besides, the cure-markedly effective rate in the former was better than that in the latter significantly (P < 0.01). CONCLUSION: There exist multiple immune functional disturbance in AHSP patients. Combined treatment of YFD and transfer factor has obvious immune regulatory effect and is an effective therapy with few side-effects and low recurrence rate. PMID- 12575572 TI - [Clinical study on relationship of CD62p, CD63 and TSP to the qi-stagnation induced blood-stasis syndrome and qi-deficiency induced blood-stasis syndrome in aged patients]. AB - OBJECTIVE: To observe the relationship of platelet activation to the Qi stagnation induced blood-stasis (QSBS) or Qi-deficiency induced blood-stasis (QDBS) syndrome. METHODS: Expressions of platelet activating molecules, including alpha-granule membrane glycoprotein (CD62p), lysosomal integral membrane protein (CD63) and thrombospondin (TSP), in patients with QSBS and QDBS were determined quantitatively with flow-cytometry and specific monoclonal antibody against activated platelet. And platelet aggregation was tested simultaneously. RESULTS: CD62p, CD63 and TSP expressions in Blood-Stasis patients, both QSBS and QDBS, were higher than those in the normal control significantly (all P < 0.01); all the three expressions were higher in QSBS group than those in QDBS group (all P < 0.01), Positive correlation was shown between CD62p and CD63 (r = 0.740, P < 0.01), CD62p and TSP (r = 0.744, P < 0.01), TSP and CD63 (r = 0.635, P < 0.01), and between CD62p and ADP induced platelet aggregation (r = 0.715, P < 0.01). CONCLUSION: Platelet activation was involved in the pathogenesis and development of Blood-Stasis Syndrome, especially closely related with the QSBS Syndrome. PMID- 12575573 TI - [Effect of xiongshao capsule on vascular remodeling in porcine coronary balloon injury model]. AB - OBJECTIVE: To study the effect of Xiongshao Capsule (XS) on vascular remodeling in porcine coronary balloon injury model. METHODS: Restenosis model was established by oversized balloon injury at mid-region of the left anterior descending coronary artery. The effect of drugs on late lumen loss and vascular remodeling was evaluated with quantitative histological method by combining results from histopathological analysis and coronary arteriography. RESULTS: Vascular remodeling explained (59 +/- 20)% of late lumen loss. All drugs significantly reduced late lumen loss 4 weeks after balloon injury (P < 0.05 or 0.01). Both low-dose and high-dose XS markedly reduced late lumen loss resulting from vascular remodeling, as compared with the control group (P < 0.05 and P < 0.01 respectively). CONCLUSION: Vascular remodeling played an important role in late lumen stenosis after balloon injury of coronary artery. XS could significantly inhibit pathological vascular remodeling after balloon injury, thus reduce late lumen loss and prevent restenosis. PMID- 12575574 TI - [Effect of Chinese herbal medicine for nourishing yin and removing fire on biosynthesis, secretion and regulative mechanism of gonadotropin-releasing hormone in hypothalamus]. AB - OBJECTIVE: To elucidate the effect of Chinese herbal medicine (CHM) for nourishing Yin and removing fire on the biosynthesis, secretion and regulative mechanism of gonadotropin-releasing hormone (GnRH) in hypothalamus. METHODS: The brain slices of medial basal hypothalamus of adolescent rats, which had been fed with CHM, were incubated. The content of GnRH in incubative liquid was determined during the slices were stimulating with high KC1 to observe the change of GnRH biosynthesis from tonic secretory center of GnRH (arcuate nucleus and ventromedial nucleus) in hypothalamus. The integrated optic density of GnRH positive immunoreactive substance in preoptic area of hypothalamus was determined by immunohistochemistry and image processing to observe the change of GnRH content in pulsative secretory center of GnRH (medial preoptic nucleus) in hypothalamus. The push-pull perfusion of medial preoptic area in hypothalamus was performed. The content of GnRH in serial perfusates was determined by radioimmunoassay (RIA) to observe the change of frequency and amplitude of GnRH pulse releasing from medial preoptic area in hypothalamus. The content of aspartic acid, glutamic acid and gamma-amino butyric acid in the perfusate was determined by high performance liquid chromatography-fluorometry, and the content of beta-endorphic in the perfusate was determined by RIA to observe the change of releasing amount of exciting aminoacid neurotransmitter and beta-endorphin from pulsative secretory center of GnRH (medial preoptic area) in hypothalamus. RESULTS: CHM could markedly reduce the content of GnRH in medial basal hypothalamus (arcuate nucleus and veatromeolial nucleus) and preoptic area (meolical preoptic nucleus) of hypothalamus, and could obviously lower the frequency and amplitude of GnRH pulse releasing from medial preoptic nucleus, It also could markedly decrease the releasing amount of aspartic acid and glutamic acid, while obviously increase the releasing amount of gamma-amino butyric acid and beta-endorphin from medial preoptic area of hypothalamus. CONCLUSION: CHM could markedly reduce the activity of GnRH neurons in hypothalamus through inhibiting the releasing of central exciting aminoacid neurotransmitter and promoting the releasing of central inhibiting aminoacid neurotransmitter and beta endorphin, thereby, obviously decrease the biosynthesis and secretion of GnRH from tonic and pulsative secretory center of GnRH in hypothalamus. It could be one of chief effective mechanism of CHM in efficiently treating the idiopathic precocious puberty. PMID- 12575575 TI - [Study of effect of zhichuan capsule on airway remodeling in experimental animal model of asthma]. AB - OBJECTIVE: To elucidate the effect and possible mechanism of Zhichuan Capsule (ZCC), a Chinese herbal preparation for reinforcing Kidney and invigorating Spleen, on airway remodeling when it was used in treating asthma. METHODS: Fifty SD rats were randomly divided into five groups: the normal control group, the model group, the high-dosage ZCC group, the low-dosage ZCC group and the Becotide (Beclomethasone Dipropionate) group, 10 rats in each group. The chronic asthma model was established by repeated inhalation of ovalbumin. The changes of collagen and fibronectin (Fn) content in airway wall, inner and outer diameter as well as area of respiratory tract cavity in lung slices were measured by computerized image analysis system. RESULTS: The wall contents of collagen and Fn in airway were higher (P < 0.05 and P < 0.01), while the ratio of inner diameter/outer diameter (ID/OD) and ratio of area of airway cavity/total area of airway (CA/TA) were significantly lower (P < 0.01) in the model animals than those in the normal controls. As compared with the model group, collagen and Fn contents were lower, ID/OD and CA/TA ratio were significantly higher in high dosage ZCC group, close to normal range. In the low-dosage ZCC group, although collagen content, ID/OD and CA/TA were not different to those in the model group, but the content of Fn was significantly lower (P < 0.01). CONCLUSION: ZCC could inhibit the remodeling of airway in chronic asthma by way of reducing the precipitation of collagen and Fn, thus help the prevention and treatment of chronic bronchial asthma. PMID- 12575576 TI - [Effect of xinhe granule on vascular endothelial damage and endothelial expressed endothelin and intercellular adhesion molecule-1 in rats fed with high lipid diet]. AB - OBJECTIVE: To study the protective effect of Xinhe granule (XHG) on vascular endothelial damage in rats fed with high lipid diet. METHODS: Model of vascular endothelial damage was formed by feeding high lipid diet in rats. The model animal were divided into high dosage XHG group, low dosage XHG group, composite Salvia dripping pellet group and model control group, and a blank (normal) control group was also set up. The degree of endothelial damage and positive cell count of synthesizing and secreting endothelin (ET-1) and intercellular adhesion molecule-1 (ICAM-1) were determined by endothelial cell spreading technique and immunohistochemical technique quantitatively. RESULTS: Comparison of the degree on vascular endothelial damage and endothelial secreting ICAM-1 and ET-1 showed model control group > composite Salvia dripping pellet group > low dosage XHG group > high dosage XHG group > blank control group. XHG did not show obvious lowering action on blood lipid. CONCLUSION: XHG could inhibit the endothelial expressed ICAM-1 and ET-1 in hyperlipidemia rats, thus displaying the protective effect on vascular endothelium. PMID- 12575577 TI - [Effect of shenmai injection on neurocyte apoptosis and change of cytoplasmic calcium]. AB - OBJECTIVE: To study the neurocyte apoptosis and change of cytosolic free calcium induced by hypoxia/hypoglycemia culture in SH-SY5Y human neuroblastoma cells, and the effect of Shenmai Injection (SMI) on them. METHODS: The neurocyte apoptosis rate was analysed quantitatively by flow cytometry and cytosolic calcium was determined by measuring mean fluorescent value with Fluo-3 flurometry. RESULTS: Hypoxia/hypoglycemia could induce neurocyte apoptosis and increase the cytosolic calcium in SH-SY5Y cells. SMI could inhibit the SH-SY5Y cell apoptosis and lower the concentration of cytosolic free calcium. CONCLUSION: Apoptosis is one of the death pattern of nerve cells. SMI could obviously reduce the hypoxia/hypoglycemia induced apoptosis and calcium overload, thus protect the neurocytes. PMID- 12575578 TI - [Effect of huadai xingshen decoction on cerebral somatostatin, arginine vasopressin and somatostatin mRNA expression in vascular dementia rat model]. AB - OBJECTIVE: To study the mechanism of Huadai Xingshen Decoction (HDXSD) in treating vascular dementia (VD). METHODS: Rat model of VD was established by repeated ischemia/reperfusion and treated with HDXSD. Hydergine was taken for positive control. The content of somatostatin (SS), arginine vasopressin (AVP) and SS mRNA expression model rat's cortex and hippocampus were observed. RESULTS: The SS and AVP contents of cortex and hippocampus in model rats were decreased significantly, and the SSmRNA expression in cortex of frontal lobe reduced, too. HDXSD showed an up-regulation on the above-mentioned parameters, and the effect on SS cortex and SS mRNA expression was higher than that of hydergine. CONCLUSION: HDXSD improves intellectual impairment of VD rats by increasing the synthetic ability of SSmRNA and content of AVP, which in turn improve the neurotrophic effect and promote the restoring function of injured neuron. PMID- 12575579 TI - [Experimental study of compound salvia injection in preventing and treating chronic nephrotoxicity induced by cyclosporin A in rats]. AB - OBJECTIVE: To explore the mechanism of protective effect of Compound Salvia Injection (CSI) on experimental cyclosporin A induced nephrotoxicity. METHODS: Rats were on low-salt diet and cyclosporin A (CsA) was administered once a day through gastrogavage at dosage of 30 mg/kg.d for 28 days. Expression of the mRNA for intrarenal transforming growth factor-beta 1 (TGF-beta 1) and renin was measured by reverse transcription-polymerase chain reaction (RT-PCR). Intrarenal expression of TGF-beta 1 and Collagen IV was determined by immunohistochemical assays. The effects of CSI on these changes were also evaluated. RESULTS: Chronic CsA-induced nephropathy might be correlated to TGF-beta 1 and renin mRNA up regulation as well as matric proteins accumulation in interstitium. CSI could reduce these changes. CONCLUSION: Decreased CsA-related TGF-beta 1 and renin upregulation expression and accumulation of matrix proteins in the kidney might be related to the protective mechanism of CSI on CsA-induced chronic nephrotoxicity. PMID- 12575580 TI - [Thinking on research of TCM syndrome group in the post-gene-group era]. PMID- 12575581 TI - [Thinking and strategy on treatment of optical fundus hemorrhagic diseases]. PMID- 12575582 TI - [Studies on caulis Tripterygium wilfordii and hypothalamus-pituitary-adrenal gland]. PMID- 12575583 TI - [Progression on studies of treatment of tumor with tetrandrine]. PMID- 12575584 TI - [The integrated studies on Ganoderma lucidum Kayst. Guided by Traditional Chinese medical theories]. PMID- 12575586 TI - [Observation on effect of radiotherapy and antike capsule combination therapy in treating nasopharyngeal cancer patients]. AB - OBJECTIVE: To compare the effect of radiotherapy (RT) combined with Antike capsule (AC) and RT alone in treating nasopharyngeal cancer (NPC) patients. METHODS: Eighty-nine patients with pathologically confirmed NPC (stage II-IV) were randomly divided into two groups: group A (46 cases) were treated with RT, receiving 65-70 Gy/6.5-7 weeks to nasopharynx region and the same dosage to neck region, and AC was given in combination. Group B (43 cases) received the same RT alone. The total dosis of RT for complete remission (CR) of primary nasopharyngeal tumor and neck lymph nodes, the CR rate and the changes of peripheral NK cell, T lymphocyte subsets in the two groups were compared. RESULTS: The total dosis of RT for CR in group A and B were 41.6 +/- 8.9 Gy vs 50.7 +/- 9.2 Gy for primary nasopharyngeal tumor, P < 0.05 and 47.4 +/- 10.3 Gy vs 56.2 +/- 9.7 Gy for neck lymph nodes, P < 0.05. The CR rate of primary nasopharyngeal tumor in group A and B were 93.5% and 88.4% respectively, P < 0.05. The activity of NK cell as well as T3, T4 in peripheral blood increased significantly in the group A after treatment, P < 0.05, while in group B, T3, T4 lowered significantly, P < 0.05. CONCLUSION: RT combined with AC could be helpful in elevating and promoting the remission rate of primary tumor and neck lymph nodes, and AC has some effects in improving the immune function and general condition in NPC patients during RT. PMID- 12575585 TI - [Clinical observation on effect of chemotherapy combined with Chinese medicine in treating advanced tumor patients and on immunologic parameters]. AB - OBJECTIVE: To observe the effects of chemotherapy combined with Chinese herbal medicine (Spleen-Kidney tonifying) in treating advanced tumor patients and on immune parameters. METHODS: One hundred and one advanced tumor patients were randomly divided into the treated group (54 patients) treated by Chinese medicine combined with chemotherapy, and the control group (47 patients) treated by chemotherapy alone. After being treated for 8 weeks (2 treatment courses), the changes of tumor size, body weight, Karnofsky scores, immunologic parameters, peripheral blood cells, as well as the toxic and side-effects were also studied. RESULTS: Improvement of various degree was obtained on the immunologic parameters such as CD3, CD4, CD4/CD8, NK cells, and the quality of life, as Karnofsky score in the treated group, in comparing with those in the control group the difference was significant (P < 0.05, P < 0.01). But on behalf of the toxic and side-effects in advanced patients, there was no significant difference between the two groups. CONCLUSION: Chemotherapy combined with Chinese Spleen-Kidney tonifying drugs could improve the immunologic functions in the advanced tumor patients. PMID- 12575587 TI - [Clinical observation of music therapy combined with anti-tumor drugs in treating 116 cases of tumor patients]. AB - OBJECTIVE: To observe the clinical effect of music therapy in treating tumor patients. METHODS: Music therapy combined with anti-tumor drugs, including chemotherapy and Chinese drugs, was given to 162 tumor patients according to syndrome differentiation to observe the change of self-rating depression scale (SDS), self-rating anxiety scale (SAS), minnesota multiphasic personality inventory (MMPI), Hamilton rating scale for depression (HAMD) and T lymphocyte subsets (immuno-histochemical assay), NK cell anti-tumor activity (NAG method), etc. while 46 caces didn't receive music therapy were taken as the control group. RESULTS: The scale marks of SDS and SAS of the treated group after treatment were obviously lower than that of the control group significantly (P < 0.05, P < 0.01). After treatment, the average values of MMPI on falseness (F), hypochondriasis (HS), depression (D) and psychosthenia (Pt) in the treated group were all improved (P < 0.01 or P < 0.05); but in the control group, significant difference only showed in MMPI on HS (P < 0.05). HAMD in the treated group revealed some improvement in insomnia, early awakening, daily work and interest, systemic symptoms and hypochondriasis (P < 0.05), and significant improvement in depression, difficulty in falling asleep, psychiatric anxiety and somatic anxiety (P < 0.01); while in the control group, only work interest and HS had some improvement (P < 0.05). CD8 percent was reduced in both groups after treatment (P < 0.01), but in the treated group CD3, CD4 and CD4/CD8 ratio were not significantly changed after treatment (P > 0.05); while in the control group they lowered obviously (P < 0.05). As for NK cell anti-tumor activity in the treated group before and after treatment, it was not significantly lowered (P > 0.05); while in the control group the lowering after treatment was significant (P < 0.05). CONCLUSION: Music therapy could regulate the emotion of tumor patient, optimize the emotional effect, improve the somatic symptoms, enhance the immune function, motivate the active principle and raise the self-regulating power in the body. PMID- 12575588 TI - [Clinical observation on small doses Tripterygium wilfordii polyglycoside combined with methotrexate in treating rheumatoid arthritis]. AB - OBJECTIVE: To observe the effect and adverse reaction of small doses Tripterygium wilfordii polyglycoside (TWP) combined with methotrexate (MT) in treating rheumatoid arthritis (RA). METHODS: Seventy RA patients were randomly divided into two groups, the control group (35 patients) and the TWP combined MT Group (TWPM group). Both of them were continued to use the non-steroidal anti inflammatory drugs. The control group took MT 15 mg orally, once every week; the TWPM group took TWP 10 mg orally, 3 times a day, and MT 7.5 mg orally once every week. The clinical effect and adverse reaction after treatment were evaluated. RESULTS: The markedly effective rate in the control group and the TWPM group was 28.6% and 34.3% respectively, with no significant difference (P > 0.05). Data of symptoms and signs, erythrocyte sedimentation rate (ESR), rheumatoid factor (RF) were determined respectively with significant difference (P < 0.01). The rate of adverse reaction was 20 cases-times in the control group and 8 cases-times in the TWPM group. CONCLUSION: MT combined small doses of TWP in treating RA has better effect and less adverse reactions than un-combined MT. PMID- 12575589 TI - [Clinical study on conducting physical exercise in lowering blood pressure of hypertensive patients and influencing their endocrine hormones]. AB - OBJECTIVE: To explore the mechanism of conducting physical exercise to lower blood pressure of hypertensive patients, relieve their symptoms and influence the cardiovascular endocrine hormone. METHODS: One hundred and twenty-five patients with II or III stage hypertension were divided into the experimental group and the control group and observed. The comparison of conditions before and after treatment, and between experimental and control groups was carried out. RESULTS: The effect in lowering blood pressure and relieving symptoms in the experimental group were obviously better than those in the control group, P < 0.05. Angiotensin-II (AT-II) level much reduced after the exercise experiment, P < 0.05. Peripheral renin activity (PRA) level had a tendency of reduction, but atrial natriuetic factor (ANF) and 6-keto-prostaglandin F1 alpha (6-k-PGF1 alpha) showed a tendency of increasing. CONCLUSION: Physical exercise combined with medical treatment could control the symptoms of hypertensive patients, in some of them could gradually using the way of doing physical exercise to replace the hypotensive medicament, the effect of the combined treatment can be obtained within a month. The change of cardiovascular endocrine hormone indicated that the physical exercise can be helpful in lowering blood pressure and improving blood circulation in hypertensive patients. PMID- 12575590 TI - [Effect of huancongdan capsule on lipoprotein, apolipoprotein and serum immunoglobulin in vascular dementia patients]. AB - OBJECTIVE: To probe into the effect of Huancongdan capsule (HCDC) on the vascular dementia (VD) related factors, including lipoprotein, apolipoprotein (apo) and serum immunoglobulin. METHODS: Fifty-two patients of VD were divided into the HCDC group (27 patients, treated with HCDC) and the control group (25 patients, treated with Naofukang). Changes of related parameters were observed. RESULTS: The effects in lowering triglyceride and total cholesterol and elevating high density lipoprotein cholesterol in the HCDC group were superior to those in the control group (P < 0.01 or P < 0.05), but the effect in lowering low density lipoprotein cholesterol was similar in the two groups (P > 0.05). Compared with data before treatment, in the HCDC group after treatment, the levels of serum apoA1 increased, and serum apoB100, IgA, IgG and circulating immune complex decreased significantly (P < 0.05 or P < 0.01), but the change of apoE level was insignificant (P > 0.05). CONCLUSION: Through adjusting lipid metabolism, HCDC could improve blood rheologic characteristics, promote cerebral blood flow, improve the oxygen carrying capacity of blood to prevent and treat VD. One of its mechanisms is increasing apoA1 and decreasing apoB100 levels, preventing and improving atherosclerosis and reducing damage of vascular endothelial cells, the another may be its action in improving immune function. PMID- 12575591 TI - [Clinical observation of compound salvia injection in treating mid-severe infantile hypoxic-ischemic encephalopathy]. AB - OBJECTIVE: To observe the changes of endothelin-1 (ET-1), nitrogen oxide (NO) and creatine phosphokinase BB isozyme (CK-BB) in blood and cerebrospinal fluid (CSF) in patients of infantile hypoxic-ishemic encephalopathy (HIE), and explore the efficacy of compound Salvia injection (CSI) in treating mid-severe HIE. METHODS: Sixty mid-severe infantile HIE patients were divided randomly into the treated and the control group. To the treated group CSI was added on the basis of conventional treatment, and to the control group the conventional treatment was given alone. The blood and CSF content of ET-1, NO and CK-BB at acute and convalescent stage in the two groups were determined and the therapeutic effects were compared between the two groups. RESULTS: The markedly effective rate and effective rate of the treated group was 80.0% and 93.3% respectively, while that of the control group was 66.7% and 83.3% respectively, the therapeutic effect in the treated group were obviously superior to that in the control group, the difference was significant (P < 0.05). CONCLUSION: ET-1, NO and CK-BB participated the pathological process of HIE. CSI was markedly effective in treating mid and severe HIE infants. PMID- 12575592 TI - [Relationship between endotoxemia and APACHE-III scoring and integrated therapy of Chinese and Western medicine]. AB - OBJECTIVE: To explore the relationship between endotoxemia and the severity of disease condition and therapeutic effect. METHODS: According to randomized controlled principle, the 153 acute infectious patients were divided into simple antibiotics treated group and antibiotics plus Chinese drugs combined treated group, and patients in each group were subdivided into 3 types according to acute physiology and chronic health evaluation (APACHE-III) scoring: type A (APACHE-III scoring < or = 20 points), type B (APACHE-III scoring 21-40 points) and type C (APACHE-III scoring > 40 points). The 77 cases in the simple treated group were 40 males and 37 females, aging 18-76 years, mean 46.5 +/- 27.5 years, 41 cases of type A, 28 of type B and 8 of type C, treatment course 10-14 days, mean 11.5 +/- 2.5 days. The 76 cases in the combined treated group were 39 males and 37 females, aging 18-70 years, mean 44.5 +/- 25.5 years, 37 of type A, 30 of type B and 9 of type C, treatment course 10-14 days, mean 10.5 +/- 2.5 days. Limulus test was used to determine the endotoxin content in peripheral blood of patients, and further analysis on the relationship between endotoxemia and APACHE-III scoring was conducted. RESULTS: Acute severe infectious patients whose APACHE-III scoring > 20 points occurred endotoxemia (P < 0.05), and the condition of disease was positively related to the APACHE-III scoring (r = 0.718, P < 0.05). Chinese drugs plus antibiotics can obviously alleviate endotoxemia (P < 0.05) and improve the prognosis of patients. CONCLUSION: Endotoxemia can serve as a referential parameter for predicting the severity of disease. Integrated therapy of Chinese and western medicine in treating bacterial infection revealed better results than that of antibiotics solely. PMID- 12575593 TI - [Effect of yiqi jianpi drugs on apoptosis and relevant gene expression in cultured vascular smooth muscle cell]. AB - OBJECTIVE: To explore the effect of Yiqi Jianpi (YQJP, supplementing Qi and invigorating Spleen) recipe on vascular smooth muscle cells (VSMC) apoptosis and it's mechanisms. METHODS: VSMC apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nicked labeling (TUNEL) analysis, transmission electron microscope (TEM), flow cytometry and DNA agarose electrophoresis. The expression activity of bcl-2, c-myc and p53 genes were determined using Northern blotting. RESULTS: The typical characteristics of apoptotic VSMC was observed following treatment by the YQJP drug serum. The DNA extracted from VSMC revealed a ladder pattern in electrophoresis. Flow cytometric analysis revealed G0/G1 stage cell blocking phenomena, the hypodiploid apoptotic cell increased, displayed typical peak of apoptosis, the rate of cell apoptosis and YQJP serum concentration were in a dose-dependent manner. The result of hybridization showed that 30% of YQJP serum could inhibit apoptotic gene bcl-2 expression, upregulated the level of apoptogenous gene c-myc and p53 mRNA. CONCLUSION: VSMC apoptosis could be induced by YQJP recipe and its mechanism might be through affecting the apoptosis related gene expression activity. PMID- 12575594 TI - [Effect of ligustrazine peritoneal dialysate on CD40 expression of rat peritoneal mesothellial cells]. AB - OBJECTIVE: To explore the effect of Ligustrazine (LGZ) affecting the expression of CD40 on mesothelial cells (MC). METHODS: Rat's peritoneal MC isolated from peritoneal cavity were cultured and maintained under the defined conditions in vitro. They were cultured with 4.25% peritoneal dialysis solution (PDS), 4.25% PDS added LGZ (40 mg/L), 4.25% PDS added IFN-gamma (100 u/ml) and 4.25% PDS added IFN-gamma and LGZ for 30 min, the DMEM/F12 cultural medium was taken as control. The expression of CD40 on MC was detected by reverse transcript polymerase chain reaction (RT-PCR) and flow cytometry analysis. RESULTS: MC cultured in vitro expressed CD40 markedly, the expression of CD40 mRNA and its protein was markedly up-regulated following stimulation with 4.25% PDS or IFN-gamma and 4.25% PDS. LGZ (40 mg/L) could reduce the expression of CD40 mRNA and its protein significantly as it was added in 4.25% PDS and 4.25% PDS with IFN-gamma. CONCLUSION: The PDS stimulates the expression of CD40 on MC. LGZ has the function of down-regulating the expression of CD40 on MC. During long-term continuous ambulatory peritoneal dialysis, it might be helpful to alleviate chronic inflammatory reaction by adding LGZ into standard PDS, therefore prevent or retard the occurence and development of peritoneal fibrosis. PMID- 12575595 TI - [Experimental study on effect of xinfeng capsule in treating rats' adjuvant arthritis on ultrastructure of synoviocyte and splenic lymphocyte]. AB - OBJECTIVE: To observe the effect of Xinfeng capsule (XFC) on the ultrastructure of synoviocyte and splenic lymphocyte in rat adjuvant arthritis (AA) model. METHODS: Sixty rats were randomly divided into normal control group, model group, XFC group and Tripterygium wilfordii polycoside (TP) group, 15 in each. Complete adjuvant 0.1 ml was injected into right posterior metatarsus of rats subcutaneously to induce inflammation, except that in the normal control. The ultrastructural change including mitochondria swelling, vacuolation and ridge pathologic change of synoviocyte and splenic lymphocyte was observed by transmission electron microscopy (TEM). The mitochondria lesion rate (MLR) of each group was then calculated. RESULTS: The swelling degree of right posterior metatarsus in XFC and TP group obviously lowered after treatment (P < 0.05). MLR of synoviocyte in the XFC group and the TP group after treatment was also obviously lower than that in the model group (P < 0.05). MLR of splenic lymphocyte in the XFC group reduced markedly (P < 0.05) after treatment, while in the TP group, the change was insignificant (P > 0.05). In comparison with TP group, the reduction of both MLR of synoviocyte and splenic lymphocyte after treatment in the XFC group was more significant (P < 0.05). CONCLUSION: XFC could lower the degree of swelling in AA rats like TP did, but XFC could improve ultrastructural change of rat's synovicocyte and splenic lymphocyte better than that of TP, which was likely the morphological basis of XFC's therapeutical effect. PMID- 12575596 TI - [Using chengqi prescription to prevent hepatic ischemia/reperfusion injury after liver transplantation in rats]. AB - OBJECTIVE: To study the effect of Chengqi prescription in preventing hepatic ischemia/reperfusion injury after liver transplantation in rats. METHODS: Rat in situ liver transplantation model was used, donor liver was preserved in Ringer's solution for 4 hrs under 4 degrees C, then divided randomly into Chinese medicine group (Chengqi prescription), polymyxin B group, mannitol group, normal saline group and sham operation control group, each group further subdivided into two subgroups: 1 hr group and 4 hrs group after transplantation. The serum ALT and liver injury index was determined. RESULTS: In preventing hepatic ischemia/reperfusion injury after liver transplantation, the Chengqi prescription was polymyxin B showed more effective than that of mannitol and normal saline, and the effect of Chengqi prescription was higher than that of polymyxin B (P < 0.05). CONCLUSION: Chengqi prescription was effective in preventing hepatic ischemia/reperfusion injury after liver transplantation in rat. PMID- 12575597 TI - [Comparison between therapeutic effects of transcutaneous electrical nerve stimulation with the frequency of 2 Hz and 100 Hz on Chronic inflammatory pain in rats]. AB - OBJECTIVE: To observe the effects of repeated transcutaneous nerve stimulation (TENS) with low frequency (2 Hz) and high frequency (100 Hz) on the chronic inflammatory pain. METHODS: The complete Freund's adjuvant was used to make the monoadjuvant arthritis as a chronic inflammatory pain model. The model animals were divided into 3 groups, the 100 Hz TENS group, the 2 Hz TENS group and the control group. The percentage increase of body weight, circumference of ankle, score of flexion pain test and score of extension pain test were used to evaluate the therapeutic effect of TENS. RESULTS: (1) Body weight increasing rate in the 100 Hz TENS group was markedly higher than that in the 2 Hz TENS group and the control group. (2) Circumference of rats' ankle in the 100 Hz TENS group was smaller than that in the 2 Hz TENS and the control group. (3) Repeated 100 Hz TENS showed more effective on pain relief than that of 2 Hz TENS. CONCLUSION: TENS has therapeutic effects on chronic inflammatory pain in rats, and different frequencies yielded different results. PMID- 12575598 TI - [Current status of herbal medicine in USA and its development]. PMID- 12575599 TI - [Progression on studies of tumor prevention and treatment by polysaccharides of Chinese herbal medicines]. PMID- 12575600 TI - [Problems on development and clinical trial of new drugs]. PMID- 12575601 TI - [Effect of combined drug-acupuncture anesthesia on hypothalamo-pituitary adrenocortical axis response and glucose metabolism in open-heart surgery patients]. AB - OBJECTIVE: To study and evaluate the effect of electroacupuncture (EA) on hypothalamo-pituitary-adrenocortical (HPA) axis response and glucose metabolism in open-heart surgery patients undergoing cardiopulmonary bypass (CPB). METHODS: Thirty patients with atrial septal defect were divided into 3 groups, the general anesthesia group (A), the acupuncture anesthesia group (B) and the general anesthesia combined EA group (C). Peripheral blood samples were collected at the time before anesthesia, before CPB and 30 min after CPB to determine plasma beta endorphin, adrenocorticotropic hormone (ACTH), serum cortisol and blood glucose. RESULTS: Plasma beta-endorphin, ACTH and serum cortisol increased significantly in Group A and B 30 min after CPB but without significant change in Group C. Blood glucose increased in all the 3 groups 30 min after CPB but the increment in Group C was the lowest. CONCLUSION: Group A and B showed no significant inhibitory effect on HPA axis response and glucose metabolism in open-heart surgery patients undergoing CPB, but Group C could obviously inhibit those responses. Assessment from the viewpoint of stress, the general anesthesia combined with EA is the better approach for anesthesia in open-heart surgery with CPB. PMID- 12575602 TI - [Clinical study on treatment of female idiopathic precocious puberty with combined therapy of Chinese medicine and megestrol acetate]. AB - OBJECTIVE: To find effective therapeutic approach for treating true idiopathic precocious puberty suitable to our national condition and different from gonadotrophin releasing hormone agonist. METHODS: One hundred and six girls with idiopathic precocious puberty were divided into 3 groups. The 51 girls in the combination therapy group were treated with megestrol acetate (MA) and Chinese medicine for nourishing Yin and removing Fire, the 35 girls in the MA treated group were treated with MA alone and the other 20 girls were given no treatment at all as control. Luteinizing hormone releasing hormone (LHRH) stimulating test was performed before and after treatment, and size of uterus and ovary, linear growth rate, X-ray bone age measurement and final height prediction were also observed simultaneously. RESULTS: After being treated with combination therapy for 2.7 years in average, in the combination therapy group, the luteinizing hormone peak value of LHRH stimulating test was reduced from 48.5 +/- 37.1 IU/L to 12.2 +/- 9.3 IU/L (P < 0.001), size of uterus and ovary decreased, secondary sexual characteristics regressed, the bone age difference/chronological age difference value (delta BA/delta CA) reduced from 1.35 +/- 0.64 to 0.65 +/- 0.36(P < 0.001), and predictive final height increased from 153.3 +/- 3.6 cm to 158.5 +/- 4.3 cm (P < 0.001). CONCLUSION: Combination therapy could not only modulate the function of hypothalamic-pituitary-ovarian axis and the development of internal genitalia, but also could slow down skeletal growth, delay skeletal maturation, and thereby prevent premature epiphyseal fusion and increase the final height of patients. PMID- 12575603 TI - [Clinical study on regulatory effect of liandai Capsule on p21ras and mutant p53 protein expression in gastrointestinal neoplasm]. AB - OBJECTIVE: To study the regulatory effect of Liandai Capsule on p21ras and mutant p53 protein expression in gastrointestinal (GI) neoplasm. METHODS: Forty-five patients of GI neoplasm in middle or advanced stage were enrolled and divided randomly into the treated group and the control group to observe the changes of their serum p21ras and mutant p53 protein expression as well as their quality of life (QOL) before and after treatment. RESULTS: The content of p21ras and mutant p53 protein expression in the treated group decreased significantly and QOL improved after treatment, P < 0.05 or P < 0.01, while these changes in the control group were not shown. CONCLUSION: Liandai Capsule has regulatory effect on p21ras and mutant p53 protein expression in GI neoplasm. PMID- 12575604 TI - [Effect of tuoli xiaodu powder on cell-mediated immunofunction in liver carcinoma patients before and after surgical operation]. AB - OBJECTIVE: To explore the regulatory effect of Tuoli Xiaodu Powder (TLXDP) on cell mediated immunofunction (T-lymphocytes subsets) in liver carcinoma patients. METHODS: Thirty-eight patients with operable liver carcinoma were divided randomly into 2 groups. TLXDP was given to the 20 cases in Group I before and 7 days after surgical operation, but not given to the 18 cases in Group II. T lymphocyte subsets (CD3, CD4 and CD8 by monoclonal antibody labelled direct immune rosette method) and liver function, aspartate aminotransferase (AST), alanine transaminase (ALT), direct and indirect bilirubin (DBIL and IBIL) were determined before and after treatment. RESULTS: T-lymphocyte subsets elevated and liver function enhanced significantly in Group I after treatment, but unchanged in Group II. CONCLUSION: Administration of TLXDP before and after surgical operation could enhance the cell-mediated immunofunction and improve the liver function significantly in patients with liver carcinoma. PMID- 12575605 TI - [Effect of relieving blood stasis, strengthening spleen and soothing liver therapy in improving hepatic function in patients after liver-carcinomectomy]. AB - OBJECTIVE: To improve hepatic function with Chinese herbal medicine (CHM) in patients after liver-carcinomectomy. METHODS: One hundred and twenty patients were divided into the CHM group (61 patients) and the control group (59 patients). CHM therapy for relieving blood stasis, invigorating Spleen and soothing Liver was given to the CHM group after liver-carcinomectomy continuously for 5-6 weeks, and hepatic function was examined in all patients 8-9 weeks after the operation. RESULTS: The hepatic function indexes, including alanine transaminase (ALT), aspartate aminotrasferase (AST), serum albumin (ALB), gamma glutamyl transpeptidase (GGTP), total bilirubin (TBIL) and direct bilirubin (DBIL), were improved in the CHM group more significantly than those in the control group, all P < 0.05. CONCLUSION: Relieving blood stasis, strengthening Spleen and soothing Liver therapy has good effect in improving hepatic function after liver-carcinomectomy. PMID- 12575606 TI - [Clinical study on effect of sini decoction on ischemia/reperfusion injury by Holter monitoring in patients with acute myocardial infarction treated with thrombolytic therapy]. AB - OBJECTIVE: To observe the clinical effect of Sini Decoction (SND) on ischemia/reperfusion injury in acute myocardial infarction (AMI). METHODS: Randomized case-control clinical trial was conducted to observe the change of Holter monitoring in 22 cases of AMI treated with thrombolytic therapy before and after treatment. RESULTS: The lasting time of acute ST segment, total burden of myocardial infarction, QRS score, QT dispersion and occurrence of reperfusion arrhythmia in patients received SND treatment were lower than those untreated with SND (P < 0.05). CONCLUSION: SND is helpful in improving reperfusion injury of thrombolytic therapy in AMI patients. PMID- 12575607 TI - [Study on effect of Astragalus injection in treating congestive heart failure]. AB - OBJECTIVE: To observe the clinical efficacy and side-effects of Astragalus Injection (AI) in treating congestive heart failure (CHF). METHODS: Eighty-three patients of CHF with heart function of II-IV grade assessed by NYHA (New York Heart Association) classification were randomly divided into 2 groups. The 42 patients in the treated group were treated with AI 40 ml (equivalent to 80 g crude drug) by adding in 5% glucose solution 500 ml for intravenous dripping, once a day and the 41 patients in the control group were treated by nitrolingual injection 15 mg by adding in 5% glucose solution 500 ml for intravenous dripping once a day. The therapeutic course in both groups was 2 weeks and the patients were followed-up for 1-6 months. RESULTS: The clinical heart function improvement rate and the total effective rate in the treated group after 1 month treatment were 26.2% and 78.6%, and after 6 months were 34.2% and 81.6% respectively, which were superior to those in the control group significantly (P < 0.05 or P < 0.01). The levels of left ventricular ejection fraction (LVEF), fractional shortening of left ventricular short axis (FS), the ratio of maximum blood flow between the advanced and early atrial systole (E/A), stroke volume (SV), cardiac output (CO) and the cardiac index (CI) were all improved in both groups (P < 0.01 or P < 0.05), but better improvement was shown in the treated group. Follow-up study showed that the incidence of cardiac event was lower in the treated group than that in the control group (P < 0.05). CONCLUSION: AI can be took as one of the important auxiliary drugs for treatment of CHF especially in severe cases. PMID- 12575608 TI - [Effect of Astragalus saponin on vascular endothelial cell and its function in burn patients]. AB - OBJECTIVE: To study the protective effect of Astragalus saponin (AS) on vascular endothelial cell (VEC) in burn patients. METHODS: Eighty burn patients were divided randomly into 2 groups, the AS group and the burn control group. Blood level of circulatory endothelial cell (CEC), endothelin (ET), nitric oxide (NO), lactate dehydrogenase (LDH) and alanine transaminase (ALT) were determined before treatment and 1 day, 5, 10 and 21 days after treatment, and compared with those in 10 healthy persons as a normal control group. RESULTS: All the parameters determined in the AS group were significantly higher than those in the normal control group. But as compared with the burn control group, the parameters in the AS group after treatment were all lower significantly. CONCLUSION: AS has definite protective effect on VEC from injury by burn, it could also improve the function of myocardial and liver cell, alleviate the general inflammatory response simultaneously. PMID- 12575609 TI - [Effect of rhubarb on intestinal immune associated secretion in healthy mice and in burn mice]. AB - OBJECTIVE: To explore the therapeutic mechanism of rhubarb (sennosides) in protecting intestinal muco-membranous barrier by observing the change of intestinal immune associated secretion (IIAS) in mice before and after burn and the enhancing effect of rhubarb on it in healthy and in burn mice. METHODS: Bal b/c mice were divided into 4 groups. Group A (n = 8), untreated healthy mice; Group B (n = 11), healthy mice treated with 10% rhubarb decoction 12 ml/kg, once every 6 hrs; Group C (n = 7), the mice with 30% back full-thickness burn and no treatment was given; Group D (n = 9), the mice with 30% back full-thickness burn and treated with 10% rhubarb decoction 12 ml/kg, once every 6 hrs. The animals were killed 24 hrs after burn, the intestinal juice was collected after intestinal lavage, and centrifuged for determination of IgA, total protein, C3 and high density lipoprotein. RESULTS: IgA content in intestinal juice of Group A, B, C and D were 93.5 +/- 13.7 mg/L, 150.8 +/- 44.6 mg/L, 43.4 +/- 21.5 mg/L and 59.8 +/- 19.3 mg/L respectively, suggesting that it was significantly reduced by burn (P < 0.05), while rhubarb decoction could increase it in either healthy or burn mice (P < 0.05). The other immune associated substance, such as total protein, C3 and high density lipoprotein in intestinal juice were also increased significantly after rhubarb decoction treatment. CONCLUSION: Rhubarb could increase the reduced intestinal juice IgA content in mice caused by burn, it may be an important mechanism of rhubarb in protecting muco-membranous barrier. PMID- 12575610 TI - [Effect of xiaoyin jiedu yin on Staphylococcus aureus enterotoxin B induced serum interleukin 8 in mice]. AB - OBJECTIVE: To explore the therapeutic mechanism of Xiaoyin Jiedu Yin (XYJDY) in treating psoriasis. METHODS: Abnormal elevation of interleukin 8 (IL-8) level in mice was induced by Staphylococcus aureus enterotoxin B (SEB), then the antagonizing effects of XYJDY and Composite Qingdai Capsule were observed. RESULTS: Serum IL-8 level arose and reached the peak 3-5 hrs after SEB induction. XYJDY could antagonize the induction significantly, and the effect was more significant when large dosage was given. CONCLUSION: XYJDY, in either large or small dose, could effectively antagonize the SEB induction on IL-8. Applying with superantigen theory, this fact could be used to elucidate the mechanism of XYJDY in removing Heat, cooling blood and detoxifying at cell biologic level. PMID- 12575611 TI - [Experimental study on effect of fuzheng yiliu decoction on tumor cell cycle and telomerease]. AB - OBJECTIVE: To study the mechanism of anti-tumor effect of Fuzheng Yiliu Decoction (FZYLD). METHODS: S180 neoplasm strain was inoculated in Kunming mice to establish model of S180 solid tumor. The model animals were treated with FZYLD by gastrogavage, the cell cycle of tumor were checked up by flow cytometer and the telomerease kit was used to test telomerease activity. RESULTS: The stage G0/1 ratio of tumor cells in model animals treated with FZYLD increased, while cells of S stage decreased, with telomerease activity inhibited. These changes were different significantly from those in the model animals treated with normal saline (P < 0.001). CONCLUSION: FZYLD could block the tumor cell proliferation procedure and inhibit the DNA synthesis and duplication in tumor cell. And the suppression of telomerease activity might be one of the mechanisms affecting the tumor cell proliferation cycle. PMID- 12575612 TI - [Regulation of Chinese medicine bailong and hexamethylene bisacetamide on cyclin dependent kinase inhibitor P16 genes in human cancer cell]. AB - OBJECTIVE: To study the regulation of bailong, a Chinese herbal anticancer preparation and hexamethylene bisacetamide (HMBA) on cyclin dependent kinase inhibitor P16 (CKI-P16) genes in human cancer cells. METHODS: The expression of CKI-P16 in different human cancer cells treated by Bailong or HMBA under different condition, was examined using Northern hybridization, Western blotting assay, etc. RESULTS: After being treated by Bailong or HMBA, the P16 expression increased. This effect was closely related to co-regulation of cAMP-PKA and DAC PKC signal pathway. When PKA pathway was blocked with PKA inhibitor, the P16 expression decreased, while PKC pathway was blocked, it enhanced. CONCLUSION: (1) The low P16 expression in G1 phase of cancer cell, as compared with that in S, G2 and M phases, might be an important factor responsible to the incompetence of P16 in inhibiting effectively the malignant change of cancer cell. (2) Mechanism of Bailong and HMBA on cancer cell proliferation inhibition might be correlated with the enhancement of P16 expression in G1 phase of cancer cells. (3) Regulation and expression of Bailong and HMBA on P16 showed the common character of Chinese and western medicine in regulating cancer cells. (4) This study elucidated that upstream of P16 was related to cAMP/PKC signal pathway closely. PMID- 12575613 TI - [Influence of yangwei mixture scathe caused by free radical after radiotherapy on mice]. PMID- 12575614 TI - [Human T cell and monocyte modulating activity of Rhizoma typhonii in vitro]. AB - OBJECTIVE: To study the immunological modulating activity of Rhizoma typhonii (RT) on lymphocytes in vitro. METHODS: Effect of RT extract (RTE) on mice spleen cells and human lymphocytes proliferation was detected by sheep erythrocytes rosette forming test after lymphocytes had been isolated and purified. The activity of human killer cells and natrral killer (NK) cells was tested using 51 Cr free test. The production of cytokine of macrophages was examined using ELISA test and biological test and the phagocytic function of macrophage on tumor cell was determined using 3H incorporation assay. The chemical ingredients of RT were analyzed using chromatography and the molecular weight of their active principle was determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis technique (SDS-PAGE) method. RESULTS: RTE could markedly stimulate the mice spleen cells and human lymphocytes to proliferate in a dose-effect dependent manner; the effector cell of RTE is T cells. RTE enhanced human killer cell, the allo-antigen specific cytotoxic T lymphocytes (CTL) activity and the nonspecific activity of NK cells. Furthermore, RTE stimulated macrophage to produce tumor necrosis factor (TNF-alpha) and interleukin-1 (IL-1), and enhanced phagocytic activity of macrophage to tumor cells. The elution of RTE from chromatography showed that the active principle of RT was glycoproteins in nature and had an apparent molecular weight of 66 kDa. CONCLUSION: RTE has immunoenhancing activity to human T cell and macrophage, through stimulating the killer cell and phagocytosis of tumor cell and allo-antigen, which could be used clinically for modulating immune responses and for treating tumor and other diseases. PMID- 12575615 TI - [Researches on prevention and treatment of reperfusion injury in cerebral ischemia by Chinese herbal medicine]. PMID- 12575616 TI - [Minutes of health care, east and west, moving into the 21st century- traditional/complementary medicine]. PMID- 12575617 TI - [Summary of 5th National Conference of Integrated Traditional Chinese and Western Experimental Medicine]. PMID- 12575618 TI - Chronic cough induced by abacavir apart from a context of hypersensitivity. AB - We report the case of an HIV-infected woman, who presented with chronic and productive cough without sign of hypersensitivity (fever, cutaneous eruption, gastrointestinal disorders), while taking abacavir. All complementary exams being negative, the involvement of abacavir has been suspected. So the drug was stopped leading to a rapid disappearance of cough. It is the first report of chronic cough with abacavir apart of a context of hypersensitivity reaction. PMID- 12575619 TI - Capillary leakage syndrome: a case report and a review. AB - Capillary leakage Syndrome (CLS) is a rare clinical syndrome, that was first described in 1960, characterized by acute episodes of generalized edema, hemoconcentration, hypoproteinemia and monoclonal gammopathy, in the vast majority of cases. We describe a 39-year-old man with anasarca, bilateral pleural and pericardial effusions, ascites and diffuse alveolo-intersticial edema. Clinical and laboratory findings were consistent with an acute episode of CLS. Treatment with prednisone, furosemide and aminophylline was started, which lead to a gradual improvement in 48 hours. Pathophysiologically there is an increase in capillary permeability with the extravasation of fluid and plasmatic proteins to the extravascular space that can lead to hypovolaemic shock. In the second phase there is a reentry of the fluid overload leading to pulmonary edema. The etiology of this hyperpermeability still remains unclear. The role of cytokines has become central in the comprehension of pathophysiology of CLS. Adhesion molecules are probably also involved in the genesis of capillary leakage. CLS treatment remains empirical. However, at present it seems that the association of steroids with furosemide, aminophylline and terbutaline are capable of controlling the clinical manifestation of the acute episodes in most cases. To our knowledge no prophylatic therapy has clearly proven its efficacy. There are only a few series analyzing the long-term evolution of patients with CLS. Further studies are necessary with the objective to collect enough patients with CLS to observe natural history of the disease and evaluate the efficacy of empiric treatments. PMID- 12575620 TI - [Value of quantitative IgE measurement]. AB - The results of assay of IgE can only be expressed by a very imprecise system of classes. Quantification of the measurement of IgE by the method of the CAP-system of prognosis of allergic patients. Expression of IgE by a measurement of intensity of fluorescence allows the establishment of a decent curve for each allergen. It makes a method of predictive evaluation of risk of the allergy pathology for each allergen. It significantly improves the sensitivity, since it allows detection of IgE at levels less than 0.35 KUA/l. Thus it gives a considerable improvement for the clinician at the level of diagnosis avoiding the "gold standard" provocation test in allergy diagnosis as well at the level of prognosis. PMID- 12575621 TI - [Compendium of specific IgE values according to professional allergists]. AB - Professional allergists have made a compendium of data of specific IgE compared with clinical diagnosis. The data allow the establishment of curves of diagnostic probability and future following of patients according to the clinical plan of skin tests and assay of specific IgE. PMID- 12575622 TI - [Prevention of respiratory syncytial virus infection by SYNAGIS (palivizumab)]. AB - Infection with respiratory syncytial virus is frequent but most often benign. The serious forms of the illness, which make necessary hospitalisation or care in an intensive Care Unit, appear in infants of less than 6 weeks and especially in those with underlying pathologies, prematurity, congenital cardiopathies or chronic respiratory illnesses. Palivizumab (SYNAGIS) is mouse humanized monoclonal antibody which is used for prevention by monthly injections before and during the epidemic period. In a pivotal study performed on 1502 infants aged less than 6 months and former prematures of less than 36 weeks gestational age (GA) or aged less than 2 years and preventing a bronchopulmonary dysplasia, 1002 infants received 5 monthly injections, compared with 500 infants treated with placebo. There was a significant reduction of 55% risk of hospitalisation with VRS infections in the treated group, but no significant reduction in the number of stays in intensive care or deaths. The recommendation in France now is to use SYNAGIS in children aged less than 6 months, born with or GA of less then 32 weeks or aged less than 2 years and presenting a bronchopulmonary dysplasia. Questions remain on the cost-benefit ratio of this treatment and the favourable effects of this treatment in children who carry other chronic pulmonary or cardiac pathologies. PMID- 12575623 TI - [Skin tests for trophallergens and asthma]. AB - The place of trophallergens in the allergy investigation of asthmatic children is controversial. Asthma is only rarely the isolated manifestation of food allergy. The clinical history is essential for research of the associated signs that reveal a food allergy. In the absence of these associated signs, the presence of a positive test for trophallergens only rarely reflects a true food allergy, of which the presence can only be assured by a double blind oral provocation test. In addition, in nurslings, the presence of a positive skin test to a trophallergen indicates atopy in the infant, but is only a mediocre predictive factor of eventual asthma, in the absence of an associated clinical allergy. PMID- 12575625 TI - Clinical trials for HIV vaccine have begun. PMID- 12575624 TI - [The place of new antihistamines in allergy management. Apropos of desloratadine]. AB - Desloratadine, the active metabolite of loratadine, is a new antihistamine. Because of its anti allergy properties, desloratidine has an affinity for histamine receptors 25 to 100 times greater to those of the usual antihistamines, coupled with a capacity to inhibit the production of pro-inflammatory mediators. When evaluated in healthy volunteers, the half life of desloratadine has been estimated at 27 hours, which is comparable with a night time length of action. Many clinical studies made with patients suffering with allergic rhinitis or chronic idiopathic urticaria have shown a rapid symptom reduction, lasting 24 hours after first taking. This action was correlated with an improvement in socio professional activity, sleep and quality of life in general. In patients suffering from allergic rhinitis, rhinomanometry showed a significant improvement in nasal congestion by desloratadine. The clinical advantages of desloratadine on antihistamines taken previously were measured in a study made on almost 48,000 patients, of whom 91% found its efficacity satisfactory. By its powerful action, coupled with an excellent tolerance profile, desloratadine represents a real therapeutic advance for allergic patients. PMID- 12575626 TI - OR masks--safe practice or habit. PMID- 12575627 TI - Preventing perioperative adverse drug reactions. AB - The purpose of this article is to help nurses prevent adverse drug reactions (ADRs) in the perioperative, endoscopic, and special procedure areas. Up to 30% of hospitalized patients experience an ADR. Taking comprehensive medication histories, maintaining constant vigilance, and recognizing inherently dangerous patient/medication combinations are essential in preventing ADRs. The mechanisms of medication toxicity and identifying potentially preventable adverse reactions are discussed. Practical nursing guidelines and tips that patients can use to prevent ADRs from occurring are delineated. Not every ADR can be prevented, but informed nurses can implement measures to reduce their occurrence. PMID- 12575628 TI - Medication errors in the OR--a secondary analysis of Medmarx. AB - Although medication errors can result in serious patient complications or even death, a paucity of information regarding medication errors that occur in the OR exists. AORN and the US Pharmacopeia (USP) collaboratively conducted a secondary analysis of reports of medication errors that occurred in the OR. These reports were submitted to the USP via the Medmarx program. The findings will give perioperative clinicians further insight into the types and causes of medication errors that occur in the OR and will help them develop potential prevention strategies. PMID- 12575629 TI - Neurosurgical considerations in posttraumatic syringomyelia. AB - Syringomyelia is a progressive degenerative disorder that results in numerous life-altering symptoms. People with this disorder frequently are misdiagnosed and most likely will experience intractable pain for many years. Surgical procedures may offer some relief from symptoms through spinal decompression or cerebrospinal fluid shunting; however, these procedures often are unsuccessful in altering the course or progression of the disease process. Information on the implications of the disease offers insight that is helpful for nurses providing care to these patients. This article presents relevant clinical information and discusses surgical procedures, as well as a patient's perspective on living with this relatively unknown disorder. PMID- 12575630 TI - Using aviation safety measures to enhance patient outcomes. AB - Recent media reports have put a spotlight on the increasing number of medical errors occurring in US health care institutions. In contrast to health care's increasing error rate, the aviation industry is experiencing a decreasing error rate. Could the safety techniques used in the aviation industry be applied to health care? This article explores that question. The dynamics of the surgical suite are not unlike those of the cockpit of an airplane; therefore, perioperative services was selected to pilot test the aviation model of safety training. PMID- 12575631 TI - Women's victimization history and surgical intervention. AB - Research suggests that women who are abused use health care services at higher rates than those who are not abused. Few studies have examined the association between women's history of victimization and surgical intervention. A secondary analysis of quantitative data obtained originally from primary care practice records of 110 women was conducted. The relationship between women's history of victimization and the likelihood that these women had undergone surgery was examined statistically. Findings reveal that women who had experienced childhood abuse, abuse as an adult by an intimate partner, or both underwent a significantly higher number of surgeries and more major surgeries than women with no history of abuse. Screening for victimization history is recommended in perioperative settings to facilitate appropriate intervention. PMID- 12575632 TI - Robotic cardiac surgery. AB - The health care climate is evolving due to influences of new technology, and robotic surgery has become a part of many surgical procedures and specialties. Incorporation of robotic procedures in cardiac surgery has several recognized benefits for patient outcomes, including a smaller incision, decrease in pain, and shorter hospital stay. Increased use of robotics will influence how nurses educate and care for their patients and the types of health care options that will be offered to patients in the future. PMID- 12575633 TI - Recycling unused, opened sterile supplies. PMID- 12575634 TI - Tracking medical devices to ensure patient safety. AB - Registered nurses in perioperative settings and managers of perioperative departments must work together to implement policies and procedures to ensure compliance with these very important federal regulations. If the information is not recorded in the proper manner and shared with the manufacturer, patients' safety is at risk. Without the ability to contact physicians and patients, manufacturers cannot alert individuals appropriately if problems arise with a certain device. Tracking devices in the correct manner ensures that patients can be notified expediently. Nurses and managers should examine their current practices to ensure that they are consistent with federal regulations. A regular assessment should be conducted to ensure that tracking forms are completed in an accurate, timely manner, that permission to release a patient's social security number is obtained, and that the hospital is compliant with the FDA's most up-to date list of devices that must be tracked. All perioperative staff members must receive education about the tracking process in their particular institution and receive updates when the process or FDA regulations change. Maintain patient safety by ensuring that the medical device tracking process is followed accurately and meets federal regulations. PMID- 12575635 TI - Recommended practices for selection and use of surgical gowns and drapes. PMID- 12575637 TI - The recovery of bacteria from the handpiece of a high school telephone. AB - The purpose of the experiment reported in this paper was to study the bacteria on the public telephones at Montgomery Blair High School in Silver Spring, Maryland, to determine if there is a risk of infection to students who use the phones. Five phone handpieces from around the school--from four public phones and the principal's phone--were swabbed twice, at 7 a.m. and at 3 p.m., on November 6,2000. Three sites on each handpiece were swabbed: the mouthpiece, the handle, and the earpiece. The swabs were streaked onto media supportive of aerobic bacteria growth and incubated at 5 percent carbon dioxide for 24 and 48 hours at 37 degrees C. The plates were studied for quantitative and qualitative data. Microscopic examination of Gram-stain preparations and, in some cases, biochemical identification were performed on the bacterial isolates. Results showed an increase in the number of bacteria from morning to afternoon in specimens from 10 of the 15 observations (67 percent). Eight of these 10 observations found more than threefold increases in the number of bacteria. In the afternoon, more types of bacteria were found in eight of the 15 specimens. Only one specimen had decreases in the number and types of bacteria from morning to afternoon. None of the bacteria that were found, however, were known pathogens. The authors conclude that even though more bacteria were recovered from phones in the afternoon than in the morning, their study did not show a serious health risk to students who used the public telephones on the day of the experiment. PMID- 12575636 TI - Skills and abilities needed by environmental health science and protection professionals in the public sector. AB - The National Environmental Health Science and Protection Accreditation Council (EHAC) accredits undergraduate academic environmental health science programs in the United States. The guidelines used by EHAC include core and technical courses that students should take in order to graduate with a bachelor of science degree in environmental health science. As part of a review of the accreditation guidelines, the research reported in this paper was undertaken with support from the Centers for Disease Control and Prevention. To identify the skills and abilities needed by entry-level environmental health professionals in the public sector, the research 1) examined existing environmental health priorities in public-health agencies, 2) reviewed entry-level job postings, and 3) surveyed more than 120 public-health professionals across the country. The results suggest that students who graduate from accredited programs must be skilled interpersonal communicators with a broad base of technical knowledge. PMID- 12575638 TI - The readability and audience acceptance of printed health promotion materials used by environmental health departments. AB - A significant part of the work of an environmental health professional is the communication of information to clients, customers, and the public in the context of safety and health promotion or as an adjunct to enforcement activities. While a wide range of communication methods are available today, printed material still forms an important aspect of the communications methodology of environmental health departments. This paper raises a number of questions about the effectiveness of environmental health promotion brochures in common use in the United Kingdom and the problems that could arise from simply assuming that the brochures are conveying the intended message to the target audience. Through a series of case studies conducted in environmental health departments, evaluative data on a range of brochures were gathered in two interlinked stages: a readability test and a target-audience questionnaire survey. The sources of the brochures included the central government, charities, trade unions, and commercial enterprises; some brochures were produced "in house." Results indicated a common mismatch between the estimated reading age of the target audience and the reading age determined by the readability test; concern about the efficacy of using commercially sourced brochures carrying advertising that may conflict with advice on other environmental health issues; "in-house" brochures that appeared to optimize self-promotion rather than the conveyance of topic information; ineffective brochures used as an adjunct to enforcement activity; and the possibility that the latter could be introduced as defense evidence in related legal proceedings. Overall, the study showed that a well structured method for brochure choice and ongoing evaluation are essential tools for environmental health departments seeking to maximize their resources and effectiveness. PMID- 12575639 TI - The importance of probable cause. PMID- 12575640 TI - The best of the best--a biennial update on our basic inspection tools. PMID- 12575641 TI - Do you really know what pH is? PMID- 12575642 TI - Changes to the journal: greater value for you. PMID- 12575643 TI - [Problems of the evolutional physiology of the nervous system]. PMID- 12575645 TI - [John Newport Langley and his autonomic (vegetative) nervous system structure (to the 150th anniversary of birth)]. PMID- 12575644 TI - [Orbeli and evolutional physiology]. PMID- 12575646 TI - [Conservatism of the insulin signal system in the evolution of vertebrate and invertebrate animals]. PMID- 12575647 TI - [Actual problems of evolutional immunology]. PMID- 12575648 TI - [Differentiation of large vasopressinergic neurons and their regulation by signal molecules in ontogenesis]. PMID- 12575649 TI - [Evolution of water-salt exchange: from phenomenology to mechanisms of physiological function]. PMID- 12575650 TI - [The role of neuronal networks in the functional evolution of the mammalian cerebral cortex]. PMID- 12575651 TI - [Formation of central mechanisms of regulation for mammal movement functions in connection with embryo development]. PMID- 12575652 TI - [Development of the ontogenetic method for studying the evolution of contractile functions]. PMID- 12575653 TI - [Insects and vertebrates: similar structures in higher integrative centers of the brain]. PMID- 12575654 TI - [Reorganization of functional systems in ontogenesis]. PMID- 12575655 TI - Hanging on in tough times. Providers struggle to hold their ground. PMID- 12575656 TI - Rewriting the book on 'institution-speak'. PMID- 12575657 TI - Limit liability with lift programs. PMID- 12575658 TI - Where logic fails the test. PMID- 12575659 TI - Recording Medicare late charges. PMID- 12575660 TI - Gene symbol: GLI3. Disease: Greig cephalopolysyndactyly syndrome. PMID- 12575661 TI - Gene symbol: GLI3. Disease: Pallister-Hall syndrome. PMID- 12575662 TI - Gene symbol: FBN1. Disease: Marfan syndrome. PMID- 12575664 TI - Bibliography. Current world literature. Coronary artery surgery. PMID- 12575663 TI - Spinal cord disease in West Nile virus infection. PMID- 12575665 TI - Bibliography. Current world literature. Ischemic heart disease. PMID- 12575666 TI - Bibliography. Current world literature. Disease of the aorta, pulmonary, and peripheral vessels. PMID- 12575667 TI - Bibliography. Current world literature. Arrhythmias. PMID- 12575668 TI - Overheating of Midas Rex MRIV motor by induced current. PMID- 12575670 TI - Intraoperative monitoring with the MicroLux Camera System. PMID- 12575671 TI - Endovascular treatment for poorest-grade subarachnoid hemorrage in the acute stage: has the outcome been improved? PMID- 12575673 TI - Endovascular treatment for poorest-grade subarachnoid hemorrhage in the acute stage: has the outcome been improved? PMID- 12575674 TI - Bilateral multiple cervical root avulsions without skeletal or ligamentous damage resulting from blast injury: case report. PMID- 12575675 TI - Prevent occlusions with these flushing pointers. PMID- 12575676 TI - Image guided endoscopic transnasal removal of recurrent pituitary adenomas. PMID- 12575681 TI - New HIV test. Accurate results in 20 minutes flat. PMID- 12575685 TI - Historical perspectives of the twak rogas (Kushtha). AB - In the doctrines of Ayurveda the twak rogas are known by the general term Kushtha. Atharvaveda describes the twak roga in the name of Kilasa, palita, with the clinical picture of grey and white spots, generally from deeper layers of skin and referred the herbal remedies. Almost all the samhitas uniformly classified Kushtha as Maha Kushtha and Kshudra Kushtha, but there is difference of opinion about the number in each group. The historical perspectives of the Twak rogas (Kushtha) are being presented according to Rigveda, Atharvaveda, Charaka, Sushruta, Vagbhata, Madhavakara Todaramalla. PMID- 12575687 TI - [Cerebrovascular disorders and hypertension]. PMID- 12575688 TI - Moraxella catarrhalis. PMID- 12575691 TI - [Biological defense mechanism common to all multi-cellular organisms]. PMID- 12575692 TI - Arunadatta and Hemadri scholars of Ashtanga Hridaya. AB - Arunadatta who wrote the famous 'Sarvanga Sundara' commentary on Ashtanga Hridaya, was the son of Mrigankadatta and a multi-farious scholar having vast knowledge of several branches of ancient learnings. It is said that he was a native of Bengal and lived in the early period of 13th century, earlier to Hemadri. It is also believed that apart from the Sarvanga Sundara commentary, he wrote a commentary on Sushruta Samahita also and composed another work entitled 'Manushyala Chandrika' dealing and geology and architecture. Hemandra was an eminent scholar who wrote the famous commentary on Ashtanga Hridya entitled Ayurveda Rasayana. This commentary has commanded great respect from all the Ayurvedic scholars for being an erudite dissertation. Besides this work it is also believed, that, he wrote several other works such as Chaturvarga Chintamani, Sraddhapaddhati, Hemadriprayoya etc. PMID- 12575693 TI - A facile and high yielding synthesis of 2,2,3,4,4-d5-androsterone-beta-D glucuronide--an internal standard in dope. AB - A facile six-step synthesis of 2,2,3,4,4-d5-androsterone-beta-D-glucuronide (1) starting from epiandrosterone (2) in 63% yield is described and compared with several alternative synthetic pathways. Compound 1 can be used as an internal standard in screening procedures for anabolic steroids to monitor the hydrolysis step of the steroid glucuronides prior to gas chromatography-mass spectrometry (GC-MS) analysis. Thus, a time consuming solid-phase extraction step to remove possible hydrolysis inhibitors can be omitted. PMID- 12575694 TI - Galen's use of Aristotle. AB - The thought of Aristotle has stamped itself on the whole subsequent course of Natural philosophy including the philosophy of healing art since the 4th century B. C. He is known to be the great codifier of ancient Natural philosophy. He developed coherent theories of generation and functional morphology of animals and heredity in his treaties. Aristotle dissected about 70 species of animals and founded the basis of comparative Anatomy. PMID- 12575695 TI - The relation between Greek and Islamic materia medica. AB - Many studies have been made on the exact data of the introduction of Greek sciences to the Arabic culture. During the 8th and 9th centuries A.D. a big progress was done in the Arab-Islamic sciences, when the Caliphs of the Omayad and Abbasid dynasties invited many scientists, researchers and translators for translation of all sciences into Arabic. The Arabs paid special attention to the Greek natural sciences such as botany and pharmacy. Greek materia medica was a major common basis for Arab-Islamic medicine and pharmacy which in turn paved the way to the modern pharmaceutical therapy. PMID- 12575697 TI - [Usefulness and problems of whole body SPECT]. PMID- 12575696 TI - Bicycle racing. PMID- 12575698 TI - [Utility and problems of hybrid PET]. PMID- 12575699 TI - [Recent Progress in instrumentation and quantification by nuclear medicine imaging]. PMID- 12575700 TI - [Nuclear oncology: from diagnosis to therapy]. PMID- 12575701 TI - Hakeem Ali Gilani: a commentator of canon of Avicenna. AB - Hakim Ali Gilani was a native of Gilan. He was a distinguished physician and medical writer. He wrote commentary on canon of Avicenna. He came to India during Akbar's time and served two Mughal rulers, Akbar and Jahangir. He was praised very much by the medical historians for his vast knowledge, curiosity and intelligence. A brief account of his achievements has been given in this article. PMID- 12575702 TI - Aurangzeb's belief in divine help for cure of diseases. AB - Aurangzeb had employed the most learned and experienced physicians for himself, for the members of his palace and for his courtiers. During his reign many hospitals were established in the capital as well as in the other cities. The reign of Aurangzeb was note-worthy for the composition and compilation of medical texts. So we can say that Aurangzeb had a very scientific attitude towards the problems of health. We find, at the same time, that he attached great importance to virtuous deeds and divine help for cure of diseases. PMID- 12575703 TI - General methods for iminosugar synthesis. AB - Recent iminosugar syntheses starting both from commercially available sugars and non-glycidic precursors, are reported in this article. Exploiting carbohydrate starting materials, different synthetic approaches are described, mainly based on the introduction of an amino function in the sugar skeleton and the subsequent aminocyclization in order to generate the piperidine or pyrrolidine ring. The aminocyclization has been performed in different ways: reductive amination of azidoketones, intramolecular amino group attack on leaving groups and activated double bonds. In some cases one-pot amination and cyclization have been performed using ammonia or a primary amine and a di-functionalised sugar. Bicyclic compounds have also been obtained through these procedures. Starting from non carbohydrate precursors, the nitrogen containing cycle, already present or easily obtained by Diels-Alder reactions, has been variously functionalised and stereo differentiated by asymmetric induction or by resolution of the racemate. The syntheses of a variety of innovative structures, such as 1-N-iminosugars, iminosugars with two nitrogen atoms in the ring, iminosugars incorporating a guanidine function, imidazole-, triazole- and tetrazole-fused iminosugars, sugar like aza-, oxa-, and thio-indolizidines, are reported. Finally the synthesis of glycoconjugate-related iminosugars, where the "pseudoanomeric" centre is linked to amino acid or peptide residues, and iminosugar phosphonates and nucleosides have been reviewed. PMID- 12575704 TI - Regulation of human sexual behaviour, sex revolution and emergence of AIDS: a historical perspective. AB - No Society in the World permits fully free sexual behaviour. All societies utilize a variety of ways for regulation of sexual behaviour. This has been happening since antiquity. Culture also affects sexual behaviour. In western civilization there was a healthly outlook towards sexuality during Greco-Roman era. In Indian civilization also human sexuality was considered an inseparable part of life and was given a higher place in human life. Many treatises on human sexuality were written. Sex was considered as an art and was given an exalted status through the medium of sculpture work in temples of Konark & Khajuraho. But in Christian civilization sexual acts and related areas were considered immoral, debasing, dirty and abhoring. Sex-related ideas/thoughts were considered immoral in Churches and were given low status in society. Rapidly occurring social changes in 20th century- World wars I & II, urbanization, modernization, industrialization, women emancipation and strong reaction to unprecedented suppression of 19th century suppression of sexuality led to advent of sexual revolution in America & other western countries. Liberal-sex spread throughout the society. Sexual promiscuity, prostitution, homosexuality, group-sex were socially accepted on a wide scale. Presumably as a result of these tendencies a disease like AIDS has now spread from America to the whole world. Present article is an effort of analysis of historical perspective of this problem. PMID- 12575705 TI - Migratory birds and the spread of West Nile. PMID- 12575706 TI - Sanctity and sanity. AB - It is to be presupposed that, the more remarkable psychological characteristics of a saint were, the more they could have contributed to his/her suggestive power. Some recent neurophysiological studies of the human-brain launched the hypothesis that the ecstatic experiences, reported by so many saints, are connected with an electric micro-activity in the temporal lobes, especially the right one. However, it was not always easy to discern a mental illness from the normal variations within the limits of human health. PMID- 12575711 TI - A brief descriptive catalogue of Ayurvedic palm leaves manuscripts procured and preserved in the Indian Institute of Medicine. AB - It seems the survey for medical manuscripts has not been done thoroughly so far in India particularly in Andhra Pradesh. Though few Researchers toured many important places and collected a number of manuscripts, but still there are many villages and several families of physicians and scholars possessing many manuscripts, are not covered. One of the aims and objects of the Indian Institute of History of Medicine is to procure and study the rare and uncared medical manuscripts. There are many difficulties in this task as many of the families in possession of the manuscripts are generally not prepared to hand over the ancestry property i.e., manuscripts to any individual or institution due to their sentiments, though they are not able to preserve or make use of same properly. However, efforts were made by the Institute in this direction, visiting several families of hereditary physicians residing in different part of Andhra Pradesh, and procured 12 such valuable palm leaves manuscripts pertaining to Ayurveda and preserved in the Institute. A brief descriptive catalogue of these manuscripts is presented here. PMID- 12575712 TI - The results of a randomized study on the use of long-acting octreotide in hepatocellular carcinoma. PMID- 12575713 TI - Bibliography of the articles published in Bulletin of the Indian Institute of History of Medicine (1994-1996). PMID- 12575714 TI - Systematic review of celecoxib for osteoarthritis and rheumatoid arthritis. Celecoxib's relative gastrointestinal safety is overstated. PMID- 12575715 TI - General practitioner screening for excessive alcohol use. General practitioners' experiences are important. PMID- 12575716 TI - General practitioner screening for excessive alcohol use. Brief screening tools should be used in general practice. PMID- 12575717 TI - Prebiotic synthesis of nucleotides at the Earth orbit in presence of Lunar soil. AB - Modern studies now favor the fact that extraterrestrial organic molecules served as an important source of biological important substances on the primitive Earth. It is presumed that these space-made organic molecules could be transported safely to the Earth surface being associated with mineral grains. It is important to test whether nucleotides synthesized in Earth orbit could be protected by lunar surface regolite. The phosphorylation of adenosine, uridine and thymidine has been studied with respect of their further transformations and degradation in presence of mineral bed. After retrieval, HPLC analysis is used to identify all the mononucleotides of certain nucleosides. It has been shown, that exposure of the investigated nucleosides as dry films in space conditions in the presence of Lunar soil increases the yield of synthesized nucleotides in 1.1-3.0 times as compared with the exposure of the same samples in absence of Lunar soil. To identify and evaluate the principal source of energy in open space responsible for nucleotide synthesis reaction laboratory experiments were performed. It has been shown, that vacuum ultra violet (VUV 145 nm) radiation promotes nucleotide synthesis more effectively than ultra violet (UV 254 nm) while the presence of Lunar soil increases reaction yield in 1.5-2.0 times. Formation of 5' mononucleotides seemed to be the most effective reaction both in flight and in laboratory experiments. Protective action of lunar soil on synthesized nucleotides against UV radiation has been shown in open Space conditions. PMID- 12575718 TI - Stability of nucleic acid under the effect of UV radiation. AB - Nucleic acids (combined with protein molecules) are essential constituents of the living systems playing an important role in the early evolution of life as well. A specific feature of these molecules has been found and directly confirmed recently: under the influence of short-wavelength UV radiation bipyrimidine photoproducts (cyclobutane dimers and 6-4 bipyrimidines) are induced and the reversion of them can be provoked by the same photons. However, reversion is preferred by the shorter wavelengths. With increasing ratio of the longer wavelength components of the radiation (using artificial UV sources and solar light on the Earth's surface) the impact of the reversible photoproducts in the harmful biological effect decreases and other photoproducts are dominant. Assuming the photoinduced reactions (dimerisation and reversion) are statistical events, during the irradiation the chance for a number of nucleoprotein molecules to survive the radiation damage can be reality. The theoretical and experimental basis of these assumptions will be discussed in the case of bacteriophage T7 nucleoprotein. PMID- 12575719 TI - Survival of microorganisms in space protected by meteorite material: results of the experiment 'EXOBIOLOGIE' of the PERSEUS mission. AB - During the early evolution of life on Earth, before the formation of a protective ozone layer in the atmosphere, high intensities of solar UV radiation of short wavelengths could reach the surface of the Earth. Today the full spectrum of solar UV radiation is only experienced in space, where other important space parameters influence survival and genetic stability additionally, like vacuum, cosmic radiation, temperature extremes, microgravity. To reach a better understanding of the processes leading to the origin, evolution and distribution of life we have performed space experiments with microorganisms. The ability of resistant life forms like bacterial spores to survive high doses of extraterrestrial solar UV alone or in combination with other space parameters, e.g. vacuum, was investigated. Extraterrestrial solar UV was found to have a thousand times higher biological effectiveness than UV radiation filtered by stratospheric ozone concentrations found today on Earth. The protective effects of anorganic substances like artificial or real meteorites were determined on the MIR station. In the experiment EXOBIOLOGIE of the French PERSEUS mission (1999) it was found that very thin layers of anorganic material did not protect spores against the deleterious effects of energy-rich UV radiation in space to the expected amount, but that layers of UV radiation inactivated spores serve as a UV shield by themselves, so that a hypothetical interplanetary transfer of life by the transport of microorganisms inside rocks through the solar system cannot be excluded, but requires the shielding of a substantial mass of anorganic substances. PMID- 12575720 TI - Life under solar UV radiation in aquatic organisms. AB - Aquatic photosynthetic organisms are exposed to solar ultraviolet (UV) radiation while they harvest longer wavelength radiation for energetic reasons. Solar UV-B radiation (280-315 nm) affects motility and orientation in motile organisms and impairs photosynthesis in cyanobacteria, phytoplankton and macroalgae as measured by monitoring oxygen production or pulse amplitude modulated fluorescence analysis. Upon moderate UV stress most organisms respond by photoinhibition which is an active downregulation of the photosynthetic electron transport in photosystem II by degradation of UV-damaged D1 protein. Photoinhibition is readily reversible during recovery in shaded conditions. Excessive UV stress causes photodamage which is not easily reversible. Another major target is the DNA where UV-B mainly induces thymine dimers. Cyanobacteria, phytoplankton and macroalgae produce scytonemin, mycosporine-like amino acids and other UV absorbing substances to protect themselves from short wavelength solar radiation. PMID- 12575721 TI - Evolution of UV-B regulation and protection in plants. AB - Plants have evolved under the influence of UV-B radiation and have acquired systems for monitoring it and investing appropriate resources for protection against it, i.e., filters, quenchers of radicals and reactive oxygen species, and repair systems. An hypothesis for how plants monitor radiation has been presented. PMID- 12575722 TI - Deuterium-free water (1H2O) in complex life-support systems of long-term space missions. AB - Heavy water containing deuterium displays toxic property. It is stated that any quantity of a heavy isotope of hydrogen--deuterium--is undesirable to animals and plants. It was earlier shown by us that physical-chemical life support systems on board the "MIR" station fractionate (change) isotopes of hydrogen, oxygen and carbon. Therefore, the problem of regenerative systems in habitable space objects should include removal, from water, of a heavy stable isotope of hydrogen- deuterium. In this article we consider one method of obtaining deuterium-free water--decomposition of distillate water in an electrolyser to hydrogen and oxygen with subsequent synthesis in a catalytic or high-temperature reactor. The influence of deuterium-free water on the growth and development of Arabidopsis thaliana and Japanese quail is investigated. It is shown that with the help of the electrolysis method it is possible to fabricate water containing 80% less deuterium in comparison with SMOW. Experimentally, it is proved on a culture of Arabidopsis thaliana and Japanese quail that water with reduced contents of deuterium (80%) displays positive biological activity. PMID- 12575723 TI - Degraded EEG response of the human brain in function of gravity levels by the method of chaotic attractor. AB - The measurement of the influence of different gravity levels on the brain allows to explain how humans react to microgravity in space and to predict the adaptation capability of astronauts. Human electroencephalographic (EEG) signals were recorded during low and high gravity phases of three consecutive days of parabolic flights on the Caravelle aircraft in 1991. EEG signals were processed, using the method of correlation dimensions d of chaotic strange attractors. Results show clear differences between the three flights, with a general decrease over time in the attractor dimensions, a measure of the brain response to changing g levels. However, the dimension is not a one-to-one relation with g levels, as additional variations are observed. Two hypotheses are introduced, the "fatigue/stress" and the "g stress" hypotheses corresponding, respectively, to long-term fatigue accumulated over the three flights, and to short-term fatigue in response to change in g levels. The former explains the overall decrease of dimensions, the latter yields additional variations on shorter time scales. As the brain response degrades with time, at least six degraded modes were observed, explained by both short- and long-term fatigue. PMID- 12575724 TI - [Therapy refractory throat pain. Dissecting aneurysm of the aorta with cardiac tamponade]. PMID- 12575725 TI - [Disability compensation for deaf patients]. PMID- 12575727 TI - No double standards: scientists must obey the same ethical rules wherever they work. PMID- 12575726 TI - Cells without heartache. PMID- 12575728 TI - Race is on to stop human cloning. PMID- 12575729 TI - 'Act now' plea on bioterror threat: the Red Cross is calling for curbs on sensitive research before lethal germs fall into the wrong hands. PMID- 12575730 TI - Freedom and the right to die. PMID- 12575731 TI - Stem cells, cloning, and explornography. PMID- 12575732 TI - Dying in our society: philosophical and ethical aspects. PMID- 12575733 TI - Lectin-mediated drug delivery: discrimination between cytoadhesion and cytoinvasion and evidence for lysosomal accumulation of wheat germ agglutinin in the Caco-2 model. AB - Lectin-mediated drug delivery may become a promising strategy to improve the efficacy of poorly permeable drugs by utilising active high-capacity transport pathways of epithelial tissues. This requires the elucidation of the basic mechanisms of lectin uptake prior to their practical use. We studied the interaction between the dietary lectin wheat germ agglutinin (WGA) and Caco-2 cells (single cells and monolayers) by a newly established assay design that is able to discriminate between cellular binding and uptake as well as by confocal microscopy: (i) All binding sites available for WGA at the cell membrane were occupied within 10 min of incubation. (ii) Cytoadhesion was followed by immediate uptake. After 20 min, 60% (single cells) or 30% (monolayers) of the membrane bound lectin were internalised. However, regardless of cell arrangement, 80% of the surface bound lectin was taken up into the cells during the course of the experiment. (iii) About 50% of the internalised lectin accumulated within the lysosomes after 1 h. This was confirmed by assays in the presence of monensin, an inhibitor of endosomal acidification, and by colocalisation with lysosomal cathepsin followed by semiquantitative image analysis. Further analysis by immunocytochemistry suggested that the trans-Golgi complex and the caveoli were not involved. Due to cytoadhesion, cytoinvasion and partial lysosomal accumulation, WGA-mediated drug delivery may provide for improved intracellular availability of conjugated drugs or colloidal carrier systems. PMID- 12575734 TI - Transport of chitosan microparticles for mucosal vaccine delivery in a human intestinal M-cell model. AB - Uptake of particulate antigen carrier systems by specialized M-cells of the gut associated lymphoid tissue is still a limiting step in inducing efficient immune responses after oral vaccination. Although transport of soluble drugs over the epithelial barrier of the gut is extensively studied in vitro by using the Caco-2 cell model, this was for long time not possible for particles due to the absence of M-cells. By co-culturing Caco-2 cells with cultured human B-lymphocytes (Raji cells), cells which are morphologically and functionally similar to M-cells can be induced. This human M-cells model makes it possible to study the uptake of microparticles for oral vaccine delivery. In this way, chitosan microparticles, which have demonstrated to target the Peyer's patches efficiently in vivo, could be tested in vitro. The development of this M-cells model facilitates the optimization of the microparticles in order to target them even more efficiently to the M-cells in the gut. In this study, the integrity of the human M-cell model was investigated by determining the transepithelial electrical resistance (TEER), 14C-mannitol transport and morphology using scanning electron microscopy. The uptake of particles was investigated by measuring transport of both fluorescently labeled microspheres (Fluospheres) and chitosan microparticles using flowcytometry. No discontinuities or abnormalities could be found in the co culture. Scanning electron microscopy showed that morphologically different cells were present in the human M-cell model. Both commercially available Fluospheres (size 0.2 microm) and chitosan microparticles (size 1.7 microm) for oral vaccine delivery were transported at a significantly higher amount by the human M-cell model compared to the transport by the Caco-2 cell monoculture. Since chitosan microparticles were proven to be taken up by Peyer's patches in mice as well, this human M-cell model is able to predict the M-cell uptake of microparticles for oral vaccine delivery. This M-cell model is a new tool, which can be used to scan, develop and optimize microparticles for oral vaccine delivery. Since the M cell uptake can now be studied in vitro, the targeting of these cells can be studied more efficiently and can now be done in cells from human origin. PMID- 12575735 TI - Chemotherapeutic targeting of etoposide to regions of the brain on the basis of polyamine level. AB - The effects of etoposide on body weight, regional weights, and the concentrations of putrescine, spermidine and spermine in the cerebellum, hippocampus, corpus striatum, cortex, the combined regions of the thalamus and hypothalamus and the diencephalon of the brain were examined in rats. Etoposide seems to be a better choice for management of cortical and hippocampal tumors because it reduces polyamines, which are associated with tumor cell growth, but not for the management of tumors of the diencephalon and corpus striatum because it increases polyamines. PMID- 12575736 TI - Blood-brain barrier disruption following the internal carotid arterial perfusion of alkyl glycerols. AB - Non ionic, amphipathic molecules form vesicles and this property correlates with the disruption of membranes. In the present studies, high mM concentrations of aliphatic alcohols, 1-O-hexyldiglycerol (HDG) and 1-O-heptyltriglycerol (HTG), are shown to cause enhanced drug transport into brain via disruption of the blood brain barrier (BBB) in vivo, as determined with an internal carotid artery perfusion method. The intravenous administration of comparable concentrations of HDG or HTG caused no increase in BBB transport of drug. The enhanced transport of drug showed a dependency on molecular weight as 45 mM HTG increased the transport of sucrose, 360 Da, but did not increase the transport of arginine vasopressin (AVP), 1084 Da, although AVP transport across the BBB was increased by 80 mM HDG or HTG. Quasielastic light scattering measurements provided evidence for the formation of vesicular structures in aqueous solutions containing high mM concentrations of the HDG or HTG. In summary, these studies demonstrate BBB disruption following the internal carotid arterial infusion of high mM concentrations of membrane active alkyl glycerols. PMID- 12575737 TI - Drug targeting to the brain: transfer of picolinic acid along the olfactory pathways. AB - Picolinic acid (PA) protects against quinolinic acid- and kainic acid-induced neurotoxicity in the brain. To study the uptake of PA to the brain, we administered [3H]PA via a unilateral nasal instillation or iv injection to mice. Autoradiography demonstrated a rapid uptake of radioactivity in the olfactory nerve layer and in the ipsilateral olfactory bulb (OB) following nasal instillation of [3H]PA. After 4 h, there was a high level of radioactivity in the central parts of the ipsilateral OB and olfactory peduncle. Moreover, iv injection of [3H]PA demonstrated a selective uptake and retention of radioactivity in the OB. Gas chromatography-mass spectrometry (GC-MS) demonstrated the presence of PA and PA-glycine conjugate in the OB. In mice with reduced peripheral olfactory innervations there was a decreased uptake of [3H]PA in the OB as compared to controls suggesting that an intact olfactory neuroepithelium is a prerequisite for an uptake of PA to the OB. There is an increased interest in brain targeting of drugs with limited ability to pass the blood-brain barrier. The present results demonstrate that PA fulfils structural requirements for a transfer along the olfactory pathways to the brain. PMID- 12575738 TI - Tris lipidation--a novel drug delivery system that alters biodistribution. AB - Tris-lipidation uses Tris to produce drug-fatty acyl conjugates. Radiolabelled Tris-fatty acyl conjugates of methotrexate (MTX) were examined in biodistribution studies in BALB/c mice. Following delivery via a variety of routes, the Tris lipidated compounds demonstrated features in common with other colloid drug delivery systems. Tissues of the reticuloendothelial system localised the drug following intravenous administration, and the compounds showed prolongation at the site of injection into muscle or fatty tissue, subcutaneously or when inhaled. These findings indicate that the Tris-lipidation platform could be classed as an alternative colloid drug delivery system. PMID- 12575739 TI - Drug targeting by solid lipid nanoparticles for dermal use. AB - Long term topical glucocorticoid treatment can induce skin atrophy by the inhibition of fibroblasts. We, therefore, looked for the newly developed drug carriers that may contribute to a reduction of this risk by an epidermal targeting. Prednicarbate (PC, 0.25%) was incorporated into solid lipid nanoparticles of various compositions. Conventional PC cream of 0.25% and ointment served for reference. Local tolerability as well as drug penetration and metabolism were studied in excised human skin and reconstructed epidermis. With the latter drug recovery from the acceptor medium was about 2% of the applied amount following PC cream and ointment but 6.65% following nanoparticle dispersion. Most interestingly, PC incorporation into nanoparticles appeared to induce a localizing effect in the epidermal layer which was pronounced at 6 h and declined later. Dilution of the PC-loaded nanoparticle preparation with cream (1:9) did not reduce the targeting effect while adding drug-free nanoparticles to PC cream did not induce PC targeting. Therefore, the targeting effect is closely related to the PC-nanoparticles and not a result of either the specific lipid or PC adsorbance to the surface of the formerly drug free nanoparticles. Lipid nanoparticle-induced epidermal targeting may increase the benefit/risk ratio of topical therapy. PMID- 12575740 TI - Transfection of HEK cells via DNA-loaded PLGA and P(FASA) nanospheres. AB - HEK cells were transfected with the GFP gene using various vectors: naked DNA, lipofectamine, and both PLGA and P(FASA) plasmid-loaded nanospheres. All methods were assessed alone and with the use of chloroquine, a lysosomal enzyme inhibitor. Transfection efficiencies were determined and compared at various times post-incubation using a fluorescence standard curve. Neither naked DNA alone nor naked DNA and chloroquine were capable of transfecting cells. No differences were evident between lipofectamine with chloroquine and lipofectamine alone which transfected cells with a constant increase in efficiency up to 2 weeks. While transfection was not feasible with polymeric nanospheres alone, the addition of chloroquine allowed DNA released from nanospheres within cells to escape endosomal degradation and transfect the cells. The increase in transfection efficiency via nanospheres over time was exponential up to 1 week, as compared to the constant rate seen for the bolus-type administration of lipofectamine, indicating that nanospheres delivered DNA to the cells by a controlled release mechanism. Additionally, the effective dose delivered to cells via nanospheres was approximately 25% that of lipofectamine, indicating that transfection via PLGA and P(FASA) nanospheres might actually be more efficient. PMID- 12575741 TI - Numerical simulation of local pharmacokinetics of a drug after intravascular delivery with an eluting stent. AB - We use mathematical modelling to delineate the influence of two important factors on local pharmacokinetics of a drug delivered via an eluting stent, namely: (1) diffusional resistance of a stent coating, and (2) reversible binding of a drug to the vascular tissue. A system of differential equations that describes diffusion of the drug out of the polymeric coating of the stent into the vascular tissue and into the bloodstream, as well as reversible binding of the drug within the vascular tissue, was solved numerically and the spatial profiles of the concentration of the drug at various points of time were produced and analysed. Also, kinetic curves of the spatial average concentration of the drug within the wall were constructed, and the areas under those curves (AUC) were calculated. The simulations showed that AUC might be enhanced, if the stent is coated with a continuous layer of a drug-releasing medium with a high diffusional resistance. Both the residence time and the average concentration of the drug within the vascular wall increase in this case mainly because the coating imposes a diffusional barrier between the vascular tissue and the bloodstream, thereby reducing the wash-out. If the drug reversibly binds to the tissue, the residence time increases greatly, but the AUC for the free (unbound) drug remains unchanged, implying that the presence of the drug in the vessel is prolonged at the expense of a proportional reduction in concentration of a free drug within the tissue. These findings justify the design of eluting stents with continuous coatings with enhanced diffusional resistance and the engineering of drugs with enhanced affinity to the vascular matrix. Reversible binding to tissue may be beneficial for prolonging the presence of the drug in the target tissue, and for avoiding potential toxic peak effects of high concentrations of the free (unbound) drug. PMID- 12575742 TI - Efficiency of Dispatch and Infiltrator cardiac infusion catheters in arterial localization of nanoparticles in a porcine coronary model of restenosis. AB - Localized intramural delivery of sustained release biodegradable nanoparticles containing an antiproliferative agent could provide prolonged drug effect at the site of vascular injury that could inhibit the proliferation of smooth muscle cells and hence restenosis. The efficiency of arterial localization of nanoparticles is crucial in maximizing the drug effect in the target tissue. Therefore, the objective of the present study was to determine the comparative efficiency of the Dispatch and the Infiltrator cardiac infusion catheters to localize nanoparticles in the arterial wall. Following a standard balloon angioplasty procedure on the left anterior descending artery (LAD) in a porcine coronary model of restenosis, a suspension of nanoparticles containing a fluorescent marker was infused at the site of injury using either the Dispatch or the Infiltrator catheter. One hour following the infusion, animals were sacrificed and the nanoparticle levels in the LAD and other tissue were analyzed. The Dispatch catheter resulted in 3.3 folds greater efficiency of nanoparticle localization in the LAD than the Infiltrator catheter (309 +/- 124 vs. 93 +/- 43 microg/g of tissue, n = 6 for Dispatch and n = 5 for Infiltrator, p = 0.082, t test). It is estimated that about 2% of the arterial volume can be displaced with the nanoparticle infusion. Fluorescence microscopy of the cross-sections of the LAD revealed greater fluorescence activity in the intimal layer with both the catheters, however the arteries infused using the Dispatch catheter demonstrated relatively higher degree of fluorescence activity in the medial and adventitial layers. The transmission electron microscopy of the arterial sections demonstrated infiltration of nanoparticles in the arterial wall and the histological analysis of the sections demonstrated no apparent damage to the endothelium due to the infusion of nanoparticles. PMID- 12575743 TI - Dynamics of the VIGS-mediated chimeric silencing of the Nicotiana benthamiana ChlH gene and of the tobacco mosaic virus vector. AB - The ChlH gene, encoding for the H subunit of the magnesium chelatase enzyme, was silenced in Nicotiana bentahamiana plants by virus-induced gene silencing (VIGS), using tobacco mosaic virus (TMV) expression vector. Strong silencing of the ChlH target gene was initiated only in the apical tissues, in which the endogenous transcription level of the target gene and the level of TMV vector RNA were both very high. The virus vector was also targeted by VIGS, and its suppression was correlated with the silencing of the ChlH mRNA. In the apical tissues, the suppression of both the virus vector and the ChlH mRNA led to a reduction of the silencing pressure and, consequently, to partial recovery of the new growth from the silencing. As the virus vector and the target mRNA levels increased, silencing was reestablished. The feedback regulation system, caused by the transient increase and reduction in levels of the virus vector and ChlH mRNA, led to a fluctuation of the silenced and recovered phenotypes in the plant apex. This TMV-vector mediated silencing system differed from previously analyzed VIGS systems; although the TMV vector was initially targeted by the silencing system, it was not permanently suppressed, indicating that, in this system, TMV was able to effectively escape post-transcriptional gene silencing. PMID- 12575744 TI - Acquired resistance functions in mlo barley, which is hypersusceptible to Magnaporthe grisea. AB - Barley plants carrying a mutation in the Mlo (barley [Hordeum vulgare L.] cultivar Ingrid) locus conferring a durable resistance against powdery mildew are hypersusceptible to the rice blast fungus Magnaporthe grisea. It has been speculated that a functional Mlo gene is required for the expression of basic pathogen resistance and that the loss of Mlo function mediating powdery mildew resistance is an exception for this particular disease. Here, we report that the onset of acquired resistance (AR) after chemical as well as biological treatments is sufficient to overcome the hypersusceptible phenotype of backcross line BCIngridmlo5 (mlo) barley plants against M. grisea. Moreover, even barley plants bearing a functional Mlo gene and thus showing a moderate infection phenotype against rice blast exhibit a further enhanced resistance after induction of AR. Cytological investigations reveal that acquired resistance in mlo genotypes is manifested by the restoration of the ability to form an effective papilla at sites of attempted penetration, similarly to wild-type Mlo plants. In addition, the rate of effective papillae formation in Mlo plants was further enhanced after the onset of AR. These results demonstrate that treatments leading to the AR state in barley function independently of the Mlo/mlo phenotype and suggest that the Mlo protein is not a component of the AR signaling network. Moreover, it seems that only concomitant action of Mlo together with AR permits high level resistance in barley against blast. Higher steady state levels of PR1 and barley chemically induced mRNA correlate with higher disease severity rather than with the degree of resistance observed in this particular interaction. PMID- 12575745 TI - Sulfated fucan oligosaccharides elicit defense responses in tobacco and local and systemic resistance against tobacco mosaic virus. AB - Sulfated fucans are common structural components of the cell walls of marine brown algae. Using a fucan-degrading hydrolase isolated from a marine bacterium, we prepared sulfated fucan oligosaccharides made of mono- and disulfated fucose units alternatively bound by alpha-1,4 and alpha-1,3 glycosidic linkages, respectively. Here, we report on the elicitor activity of such fucan oligosaccharide preparations in tobacco. In suspension cell cultures, oligofucans at the dose of 200 microg ml(-1) rapidly induced a marked alkalinization of the extracellular medium and the release of hydrogen peroxide. This was followed within a few hours by a strong stimulation of phenylalanine ammonia-lyase and lipoxygenase activities. Tobacco leaves treated with oligofucans locally accumulated salicylic acid (SA) and the phytoalexin scopoletin and expressed several pathogenesis-related (PR) proteins, but they displayed no symptoms of cell death. Fucan oligosaccharides also induced the systemic accumulation of SA and the acidic PR protein PR-1, two markers of systemic acquired resistance (SAR). Consistently, fucan oligosaccharides strongly stimulated both local and systemic resistance to tobacco mosaic virus (TMV). The use of transgenic plants unable to accumulate SA indicated that, as in the SAR primed by TMV, SA is required for the establishment of oligofucan-induced resistance. PMID- 12575746 TI - Meloidogyne javanica chorismate mutase 1 alters plant cell development. AB - Root-knot nematodes are obligate plant parasites that alter plant cell growth and development by inducing the formation of giant cells for feeding. Nematodes inject secretions from their esophageal glands through their stylet and into plant cells to induce giant cell formation. Meloidogyne javanica chorismate mutase 1 (MjCM-1) is one such esophageal gland protein likely to be secreted from the nematode as giant cells form. MjCM-1 has two domains, an N-terminal chorismate mutase (CM) domain and a C-terminal region of unknown function. It is the N-terminal CM domain of the protein that is the predominant form produced in root-knot nematodes. Transgenic expression of MjCM-1 in soybean hairy roots results in a phenotype of reduced and aborted lateral roots. Histological studies demonstrate the absence of vascular tissue in hairy roots expressing MjCM-1. The phenotype of MjCM-1 expressed at low levels can be rescued by the addition of indole-3-acetic acid (IAA), indicating MjCM-1 overexpression reduces IAA biosynthesis. We propose MjCM-1 lowers IAA by causing a competition for chorismate, resulting in an alteration of chorismate-derived metabolites and, ultimately, in plant cell development. Therefore, we hypothesize that MjCM-1 is involved in allowing nematodes to establish a parasitic relationship with the host plant. PMID- 12575747 TI - TIP, a novel host factor linking callose degradation with the cell-to-cell movement of Potato virus X. AB - The cell-to-cell movement of Potato virus X (PVX) requires four virus-encoded proteins, the triple gene block (TGB) proteins (TGB25K, TGB12K, and TGB8K) and the coat protein. TGB12K increases the plasmodesmal size exclusion limit (SEL) and may, therefore, interact directly with components of the cell wall or with plant proteins associated with bringing about this change. A yeast two-hybrid screen using TGB12K as bait identified three TGB12K-interacting proteins (TIP1, TIP2, and TIP3). All three TIPs interacted specifically with TGB12K but not with TGB25K or TGB8K. Similarly, all three TIPs interacted with beta-1,3-glucanase, the enzyme that may regulate plasmodesmal SEL through callose degradation. Sequence analyses revealed that the TIPs encode very similar proteins and that TIP1 corresponds to the tobacco ankyrin repeat-containing protein HBP1. A TIP1::GFP fusion protein localized to the cytoplasm. Coexpression of this fusion protein with TGB12K induced cellular changes manifested as deposits of additional cytoplasm at the cell periphery. This work reports a direct link between a viral movement protein required to increase plasmodesmal SEL and a host factor that has been implicated as a key regulator of plasmodesmal SEL. We propose that the TIPs are susceptibility factors that modulate the plasmodesmal SEL. PMID- 12575748 TI - Partial resistance of tomato to Phytophthora infestans is not dependent upon ethylene, jasmonic acid, or salicylic acid signaling pathways. AB - We compared tomato defense responses to Phytophthora infestans in highly compatible and partially compatible interactions. The highly compatible phenotype was achieved with a tomato-specialized isolate of P. infestans, whereas the partially compatible phenotype was achieved with a nonspecialized isolate. As expected, there was induction of the hypersensitive response (HR) earlier during the partially compatible interaction. However, contrary to our expectation, pathogenesis-related (PR) gene expression was not stimulated sooner in the partially compatible interaction. While the level of PR gene expression was quite similar in the two interactions, the LeDES gene (which encodes an enzyme necessary for the production of divinyl ethers) was expressed at a much higher level in the partially compatible interaction at 48 h after inoculation. Host reaction to the different pathogen genotypes was not altered (compared with wild type) in mutant tomatoes that were ethylene-insensitive (Never-ripe) or those with reduced ability to accumulate jasmonic acid (def-1). Similarly, host reaction was not altered in NahG transgenic tomatoes unable to accumulate salicylic acid. These combined data indicate that partial resistance in tomato to P. infestans is independent of ethylene, jasmonic acid, and salicylic acid signaling pathways. PMID- 12575749 TI - Transient expression of homologous hairpin RNA causes interference with plant virus infection and is overcome by a virus encoded suppressor of gene silencing. AB - Specific post-transcriptional gene silencing (PTGS) of target genes can be induced in a variety of organisms by providing homologous double-stranded RNA (dsRNA) molecules. In plants, PTGS is part of a defense mechanism against virus infection. We have previously shown and patented that direct delivery to nontransgenic plants of dsRNA derived from viral sequences specifically interfere with virus infection. Here, we show that transient expression of constructs encoding hairpin RNA homologous to a rapidly replicating plant tobamovirus also interferes with virus multiplication in a sequence-dependent manner. A three-day lag period between delivery of hairpin RNA and virus into the same tissues completely block virus infectivity. Several hallmarks characteristic of PTGS were associated with viral interference mediated by hairpin RNA: high level of sequence identity between the hairpin RNA and the target RNA, presence of siRNAs in extracts derived from leaves infiltrated with hairpin RNA, and helper component-proteinase (HC-Pro) of potyviruses, a suppressor of PTGS, overcame interference. No evidence for a mobile silencing suppression signal induced by transient expression of HC-Pro was observed. The approach described here has the potential to be used as a versatile tool for studying the onset of PTGS in cases involving virus infection, in opposition to dsRNA-transgenic plants, which allow primarily for the study of PTGS maintenance. PMID- 12575750 TI - Genetic analysis of a pH-regulated operon from Rhizobium tropici CIAT899 involved in acid tolerance and nodulation competitiveness. AB - Rhizobium tropici CIAT899 is highly acid tolerant and a good competitor for Phaseolus vulgaris nodule occupancy at low pH values. Using Tn5 mutagenesis, we identified an operon required for acid tolerance and nodulation competitiveness. The insertion was mapped to the 5' end of atvA, encoding a product with high sequence identity to the agro-bacterial AcvB virulence protein. Complementation analyses indicated that atvA is an ortholog of acvB, both genes being required for acid tolerance. A Ser/Ala substitution in the LIPASE_SER motif of AtvA resulted in an acid sensitive Fix+ but very poorly competing strain, demonstrating that Ser-313 is essential for AtvA function. atvA is the second gene in an operon that is transcriptionally upregulated by acid shock. The acid responsive promoter was mapped to a 469-bp intergenic region located upstream of lpiA, the first gene in the operon. lpiA-like genes are found in several alpha, beta, and gamma Proteobacteria that interact with eukaryotic host cells, and they are predicted to encode membrane proteins related to the FmtC/MprF family from low G+C Firmicutes. The latter proteins are involved in resistance to cationic antimicrobial peptides. A nonpolar deletion in lpiA caused a sevenfold decrease in relative nodulation competitiveness. PMID- 12575751 TI - QTLs for tomato powdery mildew resistance (Oidium lycopersici) in Lycopersicon parviflorum G1.1601 co-localize with two qualitative powdery mildew resistance genes. AB - Tomato (Lycopersicon esculentum) is susceptible to the powdery mildew Oidium lycopersici, but several wild relatives such as Lycopersicon parviflorum G1.1601 are completely resistant. An F2 population from a cross of Lycopersicon esculentum cv. Moneymaker x Lycopersicon parviflorum G1.1601 was used to map the O. lycopersici resistance by using amplified fragment length polymorphism markers. The resistance was controlled by three quantitative trait loci (QTLs). Ol-qtl1 is on chromosome 6 in the same region as the Ol-1 locus, which is involved in a hypersensitive resistance response to O. lycopersici. Ol-qtl2 and Ol-qtl3 are located on chromosome 12, separated by 25 cM, in the vicinity of the Lv locus conferring resistance to another powdery mildew species, Leveillula taurica. The three QTLs, jointly explaining 68% of the phenotypic variation, were confirmed by testing F3 progenies. A set of polymerase chain reaction-based cleaved amplified polymorphic sequence and sequence characterized amplified region markers was generated for efficient monitoring of the target QTL genomic regions in marker assisted selection. The possible relationship between genes underlying major and partial resistance for tomato powdery mildew is discussed. PMID- 12575753 TI - Gonadal differentiation--normal and abnormal testicular development. PMID- 12575752 TI - Sry and the genetics of sex determination. PMID- 12575754 TI - Mullerian inhibiting substance: an update. AB - The decades long study of Mullerian Inhibiting Substance by numerous laboratories around the world has been driven, in large part, by pediatric surgeons and pediatric endocrinologists who have a keen interest in the molecular pathophysiology of genital tract defects that are visited upon their patients. A better understanding of the genes involved in the development of the normal reproductive tract in males and females should lead to a more rational analysis of the diseases caused by their abnormal function. Furthermore, a translation of this knowledge from the bench to the bedside may lead to clinically useful advances in the diagnosis and management of intersex patients. The molecular analyses of MIS and MIS receptor gene mutations and persistent Mullerian duct syndrome and the development of MIS ELISAs to evaluate testicular function as well as to follow the progress of gonadal tumors are several clear examples of successes over the years. It will be interesting to see what lies ahead. PMID- 12575755 TI - Micropenis: an animal model and its human correlates. PMID- 12575756 TI - Hypothalamic imprinting by gonadal steroid hormones. AB - The results of more than four decades of research on different mammalian species have established that the brain, like the rest of the reproductive system, is esentially basically female. For the male to develop structural and functional characteristics typical of his species, his brain must be exposed to testicular hormones during a critical period, or critical periods, of development. As mammals, human beings are most likely subject to this process of the hormone dependent sexual differentiation of the brain, but proving it will be difficult. Common sense ethics preclude experimental procedures such as castration of neonatal infants or exposing the female fetus to testosterone perinatally. Thus, scientists are restricted to the retrospective study of "Experiments of Nature." The results of such studies support to a degree a meaningful role of hormones in the development of the human brain. The concept of the sexual differentiation of brain structure and function has a potentially profound influence on clinical decisions with respect to sex assignment and clinical management of infants with ambiguous or poorly developed external genitalia. Because of the importance of a baby's sex in our culture, parents of such infants must be given consideration, but so should the infant whose hormonal environment prenatally may well have produced permanent changes in the structure and functional potential of his/her brain. PMID- 12575757 TI - Sexual differentiation of the human hypothalamus. AB - Functional sex differences in reproduction, gender and sexual orientation and in the incidence of neurological and psychiatric diseases are presumed to be based on structural and functional differences in the hypothalamus and other limbic structures. Factors influencing gender, i.e., the feeling to be male or female, are prenatal hormones and compounds that change the levels of these hormones, such as anticonvulsants, while the influence of postnatal social factors is controversial. Genetic factors and prenatal hormone levels are factors in the determination of sexual orientation, i.e. heterosexuality, bisexuality or homosexuality. There is no convincing evidence for postnatal social factors involved in the determination of sexual orientation. The period of overt sexual differentiation of the human hypothalamus occurs between approximately four years of age and adulthood, thus much later than is generally presumed, although the late sexual differentiation may of course be based upon processes that have already been programmed in mid-pregnancy or during the neonatal period. The recently reported differences in a number of structures in the human hypothalamus and adjacent structures depend strongly on age. Replication of these data is certainly necessary. Since the size of brain structures may be influenced by premortem factors (e.g. agonal state) and postmortem factors (e.g. fixation time), one should not only perform volume measurements, but also estimate a parameter that is not dependent on such factors as, i.e., total cell number of the brain structure in question. In addition, functional differences that depend on the levels of circulating hormones in adulthood have been observed in several hypothalamic and other brain structures. The mechanisms causing sexual differentiation of hypothalamic nuclei, the pre- and postnatal factors influencing this process, and the exact functional consequences of the morphological and functional hypothalamic differences await further elucidation. PMID- 12575758 TI - The endocrinologist's approach to the intersex patient. PMID- 12575759 TI - 5alpha-reductase-2 deficiency and complete androgen insensitivity: lessons from nature. PMID- 12575760 TI - The CHOP experience with cloacal exstrophy and gender reassignment. PMID- 12575761 TI - A framework for the ethically justified clinical management of intersex conditions. PMID- 12575762 TI - Gender identity and sex assignment: a reappraisal for the 21st century. PMID- 12575763 TI - Gender assignment and reassignment in intersexuality: controversies, data, and guidelines for research. PMID- 12575764 TI - Prenatal diagnosis and treatment of intersex states. PMID- 12575765 TI - Anatomical studies of the fetal genitalia: surgical reconstructive implications. PMID- 12575766 TI - Feminizing genitoplasty. PMID- 12575767 TI - Management of cloacal exstrophy. PMID- 12575768 TI - Total phallic construction, option to gender reassignment. PMID- 12575769 TI - Penile reconstruction with a free sensate osteocutaneous fibula flap in the surgical management of the intersex patient. PMID- 12575770 TI - Tissue engineering approaches for genital reconstruction. PMID- 12575771 TI - A celebration of twenty years of service and leadership of the Department of Surgery, University of Florida College of Medicine: a legacy in tribute to Edward M. Copeland III, M.D. PMID- 12575772 TI - A targeted approach for antiangiogenic therapy of metastatic human colon cancer. AB - The realization that the growth and spread of tumors are dependent on angiogenesis has created new avenues of research designed to help us to better understand cancer biology and to facilitate the development of new therapeutic strategies. However, the process of angiogenesis consists of multiple sequential and interdependent steps with a myriad of positive and negative regulators of angiogenesis being involved. The survival of tumors and thus their metastases are dependent on the balance of endogenous angiogenic and antiangiogenic factors such that the outcome favors increased angiogenesis. Several growth factors have been identified that regulate angiogenesis in colon cancer; the most important of these is vascular endothelial growth factor. In addition, specific integrins such as alphavbeta3 and alpha5beta1 mediate endothelial cell survival and have been shown to be overexpressed on the endothelium of colon cancer. These angiogenic mediators thus serve as targets for therapy of metastatic colon cancer and have shown promise in preclinical trials. PMID- 12575773 TI - Advances in the diagnosis and excision of breast cancer. AB - Until recently little advance in the diagnosis and excision of breast cancer has been made since the inception of needle localization breast biopsy (NLBB). Stereotactic core needle breast biopsy (SCNBB) can avoid most NLBB especially for calcifications. However, when open biopsy is necessary NLBB has been the standard of care. As many as 50 per cent of nonpalpable lesions can be seen by ultrasound (US) to avoid the unpleasantness and complications associated with NLBB. Further SCNBB leaves a blood-filled cavity that can be easily seen by US. Intraoperative US can be used to direct the excision while improving margin negativity. MRI has improved sensitivity in detecting suspicious breast lesions and techniques such as hematoma-directed US-guided breast biopsy can facilitate excision of such masses. Clearly new technologies have improved the ability to diagnosis and excise breast cancer. The onus on the surgeon is to incorporate them into standard practice to improve outcomes. PMID- 12575774 TI - Antineoplastic therapy in colorectal cancer through proteasome inhibition. AB - Upregulation of nuclear factor (NF)-kappaB is found in many forms of cancer. Activation of NF-kappaB in cancer cells by chemotherapy or radiation can blunt the ability of this therapy to induce cell death. Proteasome inhibitors stimulate apoptosis in part via prevention of NF-kappaB activation. We sought to determine whether constitutive NF-kappaB activity is present in human colon cancer. In addition we studied whether alterations of NF-kappaB activity with a proteasome inhibitor would prevent colon cancer cell growth and induce apoptosis. We demonstrated constitutive transcriptional activation of NF-kappaB in SW48 and SW116 colon cancer cells by luciferase and electromobility shift assays. This was confirmed by p65 immunostaining. This activity was further induced in the presence of chemotherapy. In colon cancer specimens constitutive activation of NF kappaB was observed in the majority of tumors. Treatment with the proteasome inhibitor (MG-132) inhibited growth and also stimulated apoptosis of colon cancer cells. We conclude that inhibition of NF-kappaB activation may be a logical therapy for certain cancers. This can be done via specific approaches on molecules necessary for keeping NF-kappaB inactivated in the cytoplasm. Other potentially useful ways to promote apoptosis in cancer cells include the utilization of proteasome inhibitors. Such inhibitors are currently being evaluated in clinical trials. PMID- 12575775 TI - Dual induction of the Epo-Egr-TNF-alpha- plasmid in hypoxic human colon adenocarcinoma produces tumor growth delay. AB - Gene therapy is a modality for the treatment of solid tumors that involves the introduction of a suicide gene into the tumor cells. Genetic radiotherapy involves the placement of a radiation-sensitive promoter upstream from a suicide gene. Because of their irregular vasculature some solid tumors are chronically hypoxic and hence are resistant to conventional treatment with chemotherapy and ionizing radiation (IR). The purpose of this study was to demonstrate that regional tumor hypoxia could be exploited to improve local tumor control. The cDNA coding the erythropoietin hypoxia-responsive element (EPO) was placed upstream from the Egr-TNF-alpha construct. WIDR human colon adenocarcinoma cells were injected into the right hind limb of nude mice and treated with Epo-Egr-TNF alpha plasmid with or without IR. Tumor volumes were measured by calipers and tumor necrosis factor (TNF)-alpha content of the tumor was determined by enzyme linked immunosorbent assay. Treatment with the combined regimen of Epo-Egr-TNF alpha plasmid + IR resulted in significant tumor growth delay. Tumor TNF-alpha content was increased by 30 per cent in the combined treatment group compared with each treatment alone. Regional tumor hypoxia can be exploited successfully to induce tumor growth delay, enhance local control, and enhance the therapeutic ratio. PMID- 12575776 TI - Neuroblastoma, apoptosis, and growth factors. AB - Neuroblastoma is the most common extracranial tumor of childhood. This tumor is associated with alterations in apoptosis and is affected by various growth factors and cytokines. In this short review we will discuss recent findings in our laboratory where we have been studying a cell culture model of neuroblastoma. PMID- 12575778 TI - Radiofrequency ablation of breast cancer. AB - Radiofrequency ablation (RFA) is currently under study as a candidate for nonsurgical removal or destruction of breast tumors. RFA of solid tumors is produced by frictional heating caused when ions in the tissue attempt to follow the changing directions of a high-frequency alternating current. The radiofrequency probe is typically placed into the tumor under ultrasound guidance, and the ablation is performed with real-time ultrasound monitoring. Preliminary studies in which RFA was followed by standard surgical resection have indicated that this technique is effective for the surgical ablation of small (< or = 2 cm) primary breast tumors. A new study at the University of Texas M.D. Anderson Cancer Center will use RFA as the sole local treatment of breast tumors < or = 1.5 cm in size to determine whether this technique is oncologically and cosmetically appropriate for the local treatment of primary breast cancer. Twenty patients with small tumors that are clearly identifiable and measurable by ultrasound will be enrolled in the study. Local control will be assessed by core and fine-needle biopsies, and long-term outcomes will be assessed using imaging studies and quality-of-life measurement scales for 5 years after treatment. PMID- 12575777 TI - Omega-3 fatty acids: implications for the treatment of tumor-associated inflammation. AB - Our in vivo and in vitro studies using omega-3 fatty acids (FA) have provided insight into the biological effects and mechanisms of their anti-inflammatory action(s). The implications for this research are profound because there are few nutritional therapies available that have the potential to be clinically effective in malignancies and other chronic inflammatory conditions as omega-3 FA. In this summary of experiments the biological effects of omega-3 FA are discussed and the potential mechanisms of action presented. PMID- 12575779 TI - The future of clinical cancer management: one tumor, one chip. AB - Recent advances in gene expression profiling technology have now made it feasible to consider using microarray technology in the routine management of the cancer patient. Microarray chips are now capable of interrogating up to 48,000 or more different genes in a single experiment using multiple platforms. Sophisticated data analysis has already demonstrated that multiple tumor types can be distinguished on the basis of their gene expression patterns. These analyses have led to the detection of new tumor markers and markers of tumor progression. Gene expression arrays have also been demonstrated to be capable of predicting the survival of patients with breast cancer, lung cancer, brain cancer, and acute lymphocytic leukemia. The future holds great promise for the rapid development of molecular medicine with diagnosis, prognosis, and even therapy being based on a single microarray chip. These developments signal a significant paradigm shift in the clinical management of human cancer. PMID- 12575780 TI - Twenty years of progress in congenital diaphragmatic hernia at the University of Florida. AB - Over the past 20 years the clinical paradigms underlying the care of children with congenital diaphragmatic hernia (CDH) have undergone profound changes. The purpose of this work is to provide an historic review of research and clinical studies related to CDH at the University of Florida (UF) and Shands Children's Hospital during the chairmanship of Edward M. Copeland, III, M.D. and to present our current clinical results. During Dr Copeland's tenure survival for newborns symptomatic with CDH treated at UF/Shands Children's Hospital has improved from less than 20 per cent to 85 per cent. Clinical observations have suggested and research studies at UF using a fetal lamb model have confirmed that fetal distress can occur late in gestation, which may predispose infants with CDH to pulmonary hypertension. However, our patient experience has confirmed that the most significant cause of mortality in human infants is not pulmonary hypertension but iatrogenic injury to their hypoplastic lungs. Strict avoidance of barotrauma in these babies has been the most important clinical advance during these two decades. Significant clinical and research problems remain including defining optimal prenatal care, management of complications during the first few weeks of life, and development of strategies to accelerate lung growth. Dramatic improvements in survival have resulted in children who manifest a number of clinical problems that were not evident when most of these patients died early in infancy. Our experience at Shand's Children's Hospital/UF indicates that feeding problems, respiratory infections, and management of subtle or overt neurologic complications may become major issues for some of these survivors and their families. PMID- 12575781 TI - Two decades of student career choice at the University of Florida: increasingly a lifestyle decision. AB - Recently, there has been a decrease in the number of graduating U.S. medical students entering surgical residency. Therefore the purpose of this study was to examine student career choice over the last two decades (1982-2002) at the University of Florida. Residency match and academic performance data for the University of Florida medical school graduates from 1982 through 2002 were obtained from the College of Medicine's Offices of Student Affairs and Education. The percentage of medical students pursuing a career in surgery rose slightly in the 1980s to peak at 22 per cent of the graduating class in 1991 and then steadily declined in the 1990s to 15 per cent in the graduating class of 2002. The percentage of women entering surgical residency has remained relatively constant (20-year mean = 6.41 +/- 0.77) despite a significant increase percentage of women in the graduating class (Class of 1983, 24%; Class of 2002, 55%). The percentage of graduating students entering a primary care residency has remained relatively constant (54.4 +/- 1.24), whereas the number of students pursuing a "lifestyle-friendly" specialty has increased substantially over the last two decades (Class of 1983, 9%; Class of 2002, 22%). In addition the mean overall grade point average of female students entering 'lifestyle-friendly' specialty residencies is significantly higher than the mean overall grade point average of all students entering a surgical residency (3.61 +/- 0.38 versus 3.48 +/- 0.35; P < 0.05). Similar to national trends the percentage of senior medical students entering surgical residency at the University of Florida is gradually declining. Although the percentage of graduating medical students entering primary care has remained relatively constant the number of students following a lifestyle friendly specialty has steadily increased. A disproportionately low number of graduating female medical students are entering surgical residency. By one measure of academic performance the quality of female graduates entering lifestyle-friendly specialty residencies is superior to that of students entering surgical residency. Lifestyle issues are increasingly important in student career choice decisions. PMID- 12575782 TI - Eyewitness to leadership. PMID- 12575783 TI - Why do breast cancer mortality rates vary across states? AB - In 2001 approximately 40,000 deaths from breast cancer will occur in the United States. Although some estimates suggest possible state-to-state variations in breast cancer mortality rates the reasons for such differences remain unknown. Our objective was to confirm whether breast cancer mortality rates are significantly different by state and to identify predictors for such variation. Administrative data from the National Center for Health Statistics (NCHS) report were used to determine statewide death rates. Analyses were similarly performed with the Surveillance, Epidemiology, and End Results (SEER) cancer database to determine predictors of high versus low mortality rates. State-level variation in breast cancer mortality rates was demonstrated in the NCHS database. A subsequent analysis of high versus low mortality states in the SEER cancer registry demonstrates that stage at presentation was a significant predictor of mortality, as "high" mortality states had more patients presenting with later-stage disease. We conclude that variations in the breast cancer mortality rates exist between states. A nearly 50 per cent increase is observed between the states with the highest and lowest mortality rates. Adjusted analyses demonstrate that stage at presentation is a more important predictor of mortality variation than treatment differences. As such breast cancer mortality rates may be best improved by targeting screening and access-to-care issues rather than treatment. PMID- 12575784 TI - Relationship of hospital volume to outcome in cardiac surgery programs in California. AB - There are more than 120 cardiac surgery programs in California. More than half perform less than 200 coronary artery bypass graft (CABG) procedures per year. This study queried the state of California discharge abstract database to analyze the relationship between annual procedural volume and outcomes of all 119 nonfederal hospitals performing cardiac surgery during 1997 through 1999. Unadjusted in-hospital mortality for CABG surgery was 3.91 per cent (923 of 23,619) in hospitals performing fewer than 200 procedures per year and 2.09 per cent (496 of 23,704) in hospitals performing > or = 500 procedures annually (P = 0.001 by Chi-square). Similar findings were noted for CABG + valve and/or aneurysm and for valve procedures. Risk-adjusted CABG data obtained from 78 of 119 hospitals found that a significant difference persisted between a volume of < 200 and > or = 200 procedures per year (P = 0.03 by t test). There was wider variation in outcome among lower-volume hospitals. However, many low-volume providers had excellent results. It is concluded that although volume is clearly related to outcome patient-related factors and process variables may be more important. Performance improvement programs should be developed to improve communication between providers with differing outcomes. PMID- 12575785 TI - Laser flap delay: comparison of Erbium:YAG and CO2 lasers. AB - The delay phenomenon has long been recognized as a powerful tool in reconstructive surgery. This phenomenon involves creating alterations in skin flap blood supply or microcirculation to increase the size of the surviving flap. In the past many reconstructive surgeons depended on surgical delay as an integral part of their surgical planning. Today surgical delay remains a reliable method for maximizing flap survival. Although surgical delay remains the gold standard many have searched for methods to create the same effect with less morbidity and reduced cost. The purpose of this study was to determine whether near-scarless delay can be performed with either the Erbium:YAG or CO2 laser using a standard McFarlane skin flap model. Four groups were identified. Surgical delay, Erbium laser delay, and CO2 laser delay groups were each compared with a nondelayed control. Each group consisted of ten Sprague-Dawley rats. On Day 0 all delay procedures were performed on the lateral periphery of the outlined dorsal skin flaps. Interruption of this lateral blood supply was accomplished by two parallel 10-cm incisions in the surgical delay group. Likewise blood supply and microcirculatory alterations were accomplished in the laser delay groups by two parallel 10-cm laser treatments. On Day 7 a 10 x 4-cm cranially based dorsal skin flap was elevated. On Day 14 flap survival was analyzed by calculating percentage flap survival. The Erbium:YAG laser delay of the McFarlane flaps resulted in an average of 32 per cent less flap loss compared with controls (P = 0.0001). The CO2 laser resulted in an average of 36 per cent less flap loss compared with controls (P = 0.0002), whereas the surgical delay group had a 23 per cent smaller flap loss (P = 0.009). There was no significant difference between any of the delay groups. These results indicated that CO2 and Erbium:YAG lasers are as effective as surgery for delay of skin flaps in the rat model. They may provide an effective and inexpensive method for near-scarless skin flap delay in humans. PMID- 12575786 TI - Management of traumatic abdominal wall hernia. AB - Traumatic abdominal wall hernia (TAWH) can occur after blunt trauma and can be classified into low- or high-energy injuries. Low energy injuries occur after impact on a small blunt object. High-energy injuries are sustained during motor vehicle accidents or automobile versus pedestrian accidents. We present six cases of high-energy TAWH cases that were treated at our trauma center. All patients presented with varying degrees of abdominal tenderness with either abdominal skin ecchymosis or abrasions, which made physical examination difficult. CT scan confirmed the hernia in each patient. All six patients had associated injuries that required open repair. The abdominal wall defects were repaired primarily. Three patients (50%) in our series developed a postoperative wound infection or abscess. Review of the literature on low-energy TAWH shows no associated abdominal injuries. In conclusion distinction between low- and high-energy injury is imperative in the management of TAWH. Hernias following low-energy injuries can be repaired after local exploration through an incision overlying the defect. TAWHs following high-energy trauma should undergo exploratory laparotomy through a midline incision. The defect should be repaired primarily and prosthetics avoided because of the high incidence of postoperative infection. PMID- 12575787 TI - Does fibrin sealant reduce drain output and allow earlier removal of drainage catheters in women undergoing operation for breast cancer? AB - Serosanguinous drainage after mastectomy and axillary lymph node dissection has traditionally been treated with the temporary use of closed suction drainage catheters. Use of drainage catheters is associated with wound infection, discomfort, nerve injury, and impaired arm movement. Commercially produced fibrin sealant has been proposed to reduce postoperative serosanguinous collections. We hypothesized that the intraoperative application of low-dose (2-5 cm3) fibrin sealant would reduce serosanguinous drainage and allow earlier removal of closed suction drainage catheters after operation for breast cancer. Fifty-five women with known breast cancer underwent either total mastectomy, modified radical mastectomy, or isolated level I and II axillary lymph node dissection. Twenty-six patients were treated with fibrin sealant and 29 served as control subjects. The application of fibrin sealant resulted in a significant reduction in overall duration catheters were needed (7 vs 8.3 days; P = 0.05). More importantly fibrin sealant reduced the time until 24-hour drain output was less than 30 cm3 (4.9 vs 6.2 days). Additionally fibrin sealant application resulted in a 60 per cent reduction in overall drainage amount after total mastectomy and a 32 per cent reduction after modified radical mastectomy. The application of fibrin sealant after axillary lymph node dissection did not decrease overall drainage amount. In conclusion fibrin sealant reduces serosanguinous drainage after total mastectomy and modified radical mastectomy and may allow earlier removal of closed suction drainage catheters. PMID- 12575788 TI - Preoperative sestamibi localization combined with intraoperative parathyroid hormone assay predicts successful focused unilateral neck exploration during surgery for primary hyperparathyroidism. AB - A retrospective review of 54 cases of primary hyperparathyroidism operated upon by five general surgeons at Santa Barbara Cottage Hospital between March 1998 and March 2001 was performed to determine whether positive preoperative sestamibi localization (PSL) of a solitary adenoma combined with intraoperative parathyroid hormone assay (IOPHA) could predict successful focused unilateral neck exploration. A solitary adenoma was found in each of 50 patients (93%). PSL for solitary adenomas had an accuracy of 87 per cent, positive predictive value (PPV) of 96 per cent, sensitivity of 90 per cent, and specificity of 50 per cent. Forty five patients (83%) achieved a 50 per cent reduction in IOPHA at 10 minutes after excision of a solitary adenoma for an accuracy of 85 per cent, PPV of 97 per cent, sensitivity of 88 per cent, and specificity of 50 per cent. All patients remain eucalcemic. The combination of PSL and IOPHA resulted in a PPV of 97.5 per cent and a sensitivity of 100 per cent. From these data we conclude that a focused unilateral neck exploration could have been performed successfully in 78 per cent of the cases. PMID- 12575789 TI - Acute infectious purpura fulminans: a 15-year retrospective review of 28 consecutive cases. AB - Acute infectious purpura fulminans (AIPF) is a rare syndrome of hemorrhagic infarction of the skin, extremity loss, and intravascular thrombosis. It progresses rapidly and is accompanied by disseminated intravascular coagulation and vascular collapse. The victims often succumb to the disease. Our objective was to investigate the clinical manifestations, outline the clinical course, and delineate factors related to mortality among the patients with AIPF. Patients diagnosed with AIPF over a 15-year period were reviewed retrospectively for patient history, comorbid conditions, progression of clinical course, and medical and surgical management. The vast majority of the patients were under the age of 7; however, the disease process can be seen in adults. The overall mortality rate was 43 per cent. The major predisposing factors were history of recent upper respiratory infection, recent surgery or childbirth, young age, and absence of a spleen. The most common clinical manifestations were skin discoloration, disseminated intravascular coagulation, fever, and septic shock. The most common bacteria cultures were Neisseria meningitidis, Hemophilus influenzae, and Streptococcus pneumoniae. There appears to be a higher mortality in patients who did not undergo a surgical intervention. Compartment syndrome needs to be evaluated early on in the presentation. Rapid diagnosis, intensive care unit management, and prompt surgical consultation and debridement may decrease the mortality. PMID- 12575790 TI - Location, location, location--the nuclear view. PMID- 12575791 TI - A fiber-FISH contig spanning the non-recombining region of the human Y chromosome. AB - Using fluorescence in-situ hybridization on interphase chromatin fibers (fiber FISH), we have constructed an overlapping fiber-FISH contig spanning the non recombining region of the human Y chromosome (NRY). We first established a standard FISH-signal pattern for a distinct panel of DNA clones on prometaphase Y chromosomes in six healthy fertile men. Clones in the panel were selected from all R-bands as well as deletion intervals 1 through 7 plus PAR1 and PAR2 of the human Y chromosome. We next used signals of these marker clones to build a fiber FISH contig for the multicopy gene families, CDY, DAZ, RBMY, TSPY and XKRY, along the NRY. Our fiber-FISH contig of human NRY may help to close the four gaps that still exist in the current physical map of the human Y chromosome. Furthermore, it provides a more complete picture with respect to the positions and arrangements of the multicopy gene families along the human NRY. PMID- 12575792 TI - Onset and sequence of RBA-band replication on the inactive X-chromosomes of cattle (Bos taurus L.), river buffalo (Bubalus bubalis L.) and goat (Capra hircus L.). AB - This study was undertaken to provide cytogenetic information about onset and sequence of RBA-band replication on the inactive X-chromosomes of cattle, river buffalo and goat. Blood cultures were synchronized overnight with thymidine after 48 hours of growth. The cell block was released with fresh medium and the cells allowed to grow in the presence of BrdU and H33258 for 1, 2, 4, 6, 8, 10, 12 and 14 hours, including 20 minutes colcemide. Results show that: (a) the onset of RBA banding replication was 12 hours before mitosis in cattle and river buffalo, 14 hours in the goat; (b) the replication process was still 'on' in cattle and river buffalo one hour before mitosis, whereas it was 'off' in the goat; consequently the length of the G2 phase was less than one hour in cattle and river buffalo and one hour or slightly longer in the goat; (c) the first band undergoing replication was identified as band Xq31 in cattle, homologous to band Xq34-36 in river buffalo and Xq24 in the goat; (d) the second replicating band was the Xp22 in cattle, homologous to band Xq21 in river buffalo and Xq34 in the goat, respectively; (e) the sequence of RBA-band replication was quite similar between cattle and river buffalo, but reversed in the goat, due to the wide chromosomal rearrangements which differentiated the X-chromosome of Caprinae from that of Bovinae. PMID- 12575793 TI - A strategy for enhancing recombination in proximal regions of chromosomes. AB - As a rule, recombination in bread wheat (Triticum aestivum L.) is low in proximal and high in distal regions of chromosomes. Recombination may be enhanced in proximal regions by using deletion (del) chromosomes deficient for a distal part of a chromosome arm. The chromosome del5BL-11 derived from Chinese Spring (CS) is missing 41% of the distal long arm. This line was made polymorphic by crossing with a stock in which chromosome 5B of CS (5B(CS)) is substituted for chromosome 5B of T. turgidum ssp. dicoccoides origin (5B(T.dic)). Three recombinant del5BL 11 (del5BL-11(rec)) lines were isolated, all resulting from localized recombination between loci Xbcd926 and XksuH1. In del5BL-11(rec), the centromere to fraction length (FL) 0.53 (C-FL0.53) segment is derived from 5B(T. dic) and the distal region of FL 0.55-0.59 is from 5B(CS). Genetic recombination for the C FL 0.53 interval was assayed in segregating progenies from 5B(CS)/5B(T.dic) and del5BL-11/del5BL-11(rec) crosses using polymorphic markers and for the FL 0.55 0.59 interval in del5BL-11/del5BL-11(rec) cross from chiasma counts. The pairing data and comparative mapping of normal 5B and del5BL-11 indicated that the increase in recombination was restricted to the FL 0.55-0.59 interval of the del5BL-11 chromosome. No significant increase in recombination in more proximal regions was observed although the order of several markers that cosegregated in the normal 5B map was resolved in the del5BL-11 map. The presented data show that recombination in proximal, usually low-recombination, regions can be increased by placing them close to the chromosome end. PMID- 12575794 TI - Chromosome arrangement and behaviour of two rye homologous telosomes at the onset of meiosis in disomic wheat-5RL addition lines with and without the Ph1 locus. AB - Fluorescence in-situ hybridization (FISH) of total genomic and repetitive DNA on microsporocytes of ditelocentric addition lines of rye 5RL in hexaploid wheat was performed to study the behaviour of the rye homologous chromosome arms in relation to centromere and telomere dynamics at premeiotic interphase and meiotic prophase I. By comparing isogenic lines with and without the Ph1 locus, we established the effect of the Ph1 gene on appearance and behaviour of the rye chromosomes. Ph1 and ph1b lines demonstrated similar premeiotic chromosome arrangement with the two rye homologues occupying separated domains despite the occurrence of centromere association. Our study confirmed that bouquet arrangement of telomeres follows the Rabl configuration. In cells displaying bouquet clustering of telomeres, centromeres of the 5RL telosomes are still at the opposite pole, suggesting anchoring of centromeres at the cytoskeleton. Once the telomeres complete clustering, the rye centromeres migrate to the telomere pole, and the rye chromosomes begin to loosen their structure. While the rye homologues in the wild-type keep separate territories in the nucleus, they become intermingled in the ph1b mutant, possibly because of their lower condensation. In a subsequent stage, the 5RL homologues appear intimately associated mainly at the distal region. Our study suggests that the lower rate of chromosome synapsis in the ph1b mutant results from abnormal chromatin decondensation and organization. PMID- 12575795 TI - The phylogeny of howler monkeys (Alouatta, Platyrrhini): reconstruction by multicolor cross-species chromosome painting. AB - We performed multidirectional chromosome painting in a comparative cytogenetic study of the three howler monkey species Alouatta fusca, A. caraya and A. seniculus macconnelli (Atelinae, Platyrrhini) in order to reconstruct phylogenetic relationships within this genus. Comparative genome maps between these species were established by multicolor fluorescence in-situ hybridization (FISH) employing human, Saguinus oedipus and Lagothrix lagothricha chromosome specific probes. The three species included in this study and previously analyzed howler monkey species were subjected to a phylogenetic analysis on the basis of a data matrix comprised of 98 discrete molecular cytogenetic characters. The results revealed that howler monkeys represent the genus with the most extensive karyotype diversity within Platyrrhini so far analyzed with high levels of intraspecific chromosomal variability. Two different multiple sex chromosome systems were identified. The phylogenetic analysis indicated that Alouatta is a monophyletic clade which can be derived from a proposed ancestral Atelinae karyotype of 2n = 62 chromosomes by a chromosome fusion, a fission, a Y-autosomal translocation and a pericentric inversion. Following these suggestions, the genus Alouatta can be divided into two distinct species groups: the first includes A. caraya and A. belzebul, the second A. s. macconnelli, A. sara, A. s. arctoidea and A. fusca. PMID- 12575796 TI - Chromatin preferences of the perichromosomal layer constituent pKi-67. AB - The proliferation-associated nuclear protein pKi-67 relocates from the nucleolus to the chromosome surface during the G2/M transition of the cell cycle and contributes to the formation of the 'perichromosomal layer'. We investigated the in-vivo binding preferences of pKi-67 for various chromatin blocks of the mitotic chromosomes from the human and two mouse species, Mus musculus and M. caroli. All chromosomes were decorated with pKi-67 but displayed a gap of pKi-67 decoration in the centromere and NOR regions. pKi-67 distribution in a rearranged mouse chromosome showed that the formation of the centromeric gap was controlled by the specific chromatin in that region. While most chromatin served as a substrate for direct or indirect binding of pKi-67, we identified three types of chromatin that bound less or no pKi-67. These were: (1) the centromeric heterochromatin defined by the alpha satellite DNA in the human, by the mouse minor satellite in M. musculus and the 60- and 79-bp satellites in M. caroli; (2) the pericentromeric heterochromatin in M. musculus defined by the mouse major satellite, and (3) NORs in the human and in M. musculus defined by rDNA repeats. In contrast, the conspicuous blocks of pericentromeric heterochromatin in human chromosomes 1, 9 and 16 containing the 5-bp satellite showed intense pKi-67 decoration. The centromeric gap may have a biological significance for the proper attachment of the chromosomes to the mitotic spindle. In this context, our results suggest a new role for centromeric heterochromatin: the control of the centromeric gap in the perichromosomal layer. PMID- 12575797 TI - Development of flow cytogenetics and physical genome mapping in chickpea (Cicer arietinum L.). AB - Procedures for flow cytometric analysis and sorting of mitotic chromosomes (flow cytogenetics) have been developed for chickpea (Cicer arietinum). Suspensions of intact chromosomes were prepared from root tips treated to achieve a high degree of metaphase synchrony. The optimal protocol consisted of a treatment of roots with 2 mmol/L hydroxyurea for 18 h, a 4.5-h recovery in hydroxyurea-free medium, 2 h incubation with 10 micromol/L oryzalin, and ice-water treatment overnight. This procedure resulted in an average metaphase index of 47%. Synchronized root tips were fixed in 2% formaldehyde for 20 min, and chromosome suspensions prepared by mechanical homogenization of fixed root tips. More than 4 x 10(5) morphologically intact chromosomes could be isolated from 15 root tips. Flow cytometric analysis of DAPI-stained chromosomes resulted in histograms of relative fluorescence intensity (flow karyotypes) containing eight peaks, representing individual chromosomes and/or groups of chromosomes with a similar relative DNA content. Five peaks could be assigned to individual chromosomes (A, B, C, G, H). The parity of sorted chromosome fractions was high, and chromosomes B and H could be sorted with 100% purity. PCR on flow-sorted chromosome fractions with primers for sequence-tagged microsatellite site (STMS) markers permitted assignment of the genetic linkage group LG8 to the smallest chickpea chromosome H. This study extends the number of legume species for which flow cytogenetics is available, and demonstrates the potential of flow cytogenetics for genome mapping in chickpea. PMID- 12575798 TI - Influences of chromosome size, gene density and nuclear position on the frequency of constitutional translocations in the human population. AB - Translocations are the most frequent chromosome structural aberration in the human population, yet little is known about their aetiology. Here, factors that might influence the occurrence of constitutional translocations in the population are examined. By analysing >10000 translocations from two large databases of cytogenetic abnormalities, chromosome size is identified as the major determinant of translocation frequency. This probably reflects the large target size for double-strand breakage and repair presented by the largest chromosomes. There is also evidence for selection against translocations that involve breakage through the most gene-dense chromosomes. Lastly, it is suggested that nuclear organization of chromosomes impinges on the frequency of translocations amongst the smallest autosomes. PMID- 12575799 TI - Spur cell anemia in alcoholic cirrhosis: cure by orthotopic liver transplantation and recurrence after liver graft failure. AB - Spur cell anemia is an acquired form of hemolytic anemia caused by a structural abnormality of red cell membranes that results in spiculated erythrocytes. These peculiarly shaped red blood cells, called acanthocytes, have a shortened survival and undergo splenic sequestration and destruction. Spur cell anemia has been known to occur in several conditions, including chronic liver disease, and more specifically in alcoholic cirrhosis. Treatment of this disorder has been disappointing and usually indicates end-stage liver disease. Liver transplantation has been reported as the most effective treatment. We herein present a case of severe spur cell hemolytic anemia that successfully reverted after orthotopic liver transplantation and recurred secondary to resumption of alcohol intake and consequent liver graft failure. This case conclusively demonstrates the association among alcoholic cirrhosis, end-stage liver disease, and spur cell hemolytic anemia. PMID- 12575800 TI - Perisplenitis as a cause of acute abdomen: a case report. AB - Splenitis can complicate blood-borne sepsis in hemodialysis patients. Symptoms include left upper quadrant pain and tenderness in addition to generalized systemic manifestation of infection. Clinical diagnosis is difficult and there is no specific investigation to confirm it. Computed tomography scan of the spleen can help in identifying a splenic abscess, rupture, or infarction. A splenectomy is the treatment of choice in splenic abscess, in splenitis to avoid spontaneous rupture, and in recurrent perisplenitis. PMID- 12575801 TI - Surgical management of a hydatid cyst of the hepatic dome ruptured into the biliary tree. AB - Hydatid disease of the liver remains an important and challenging medical problem. Although surgery is considered the treatment of choice for hydatid disease of the liver, controversies still exist about the preferred operative technique. We report the case of a patient with obstructive jaundice caused by rupture of a hydatid cyst of the hepatic dome into the biliary tract. He was managed by a new surgical approach of myoplasty of the right hemidiaphragm combined with preoperative decompression of the bile duct by an endoscopic sphincterotomy. The postoperative course of the patient was uneventful, and he remains well 3 years later. This procedure is simple, requires less time, and has the advantage of avoiding a thoracoabdominal incision, which makes any abdominal reoperation easier. PMID- 12575802 TI - Right psoas muscle/aortoiliac groove recurrence: an unusual anatomic site for progression of epithelial tumors of the appendix. AB - Appendiceal carcinoma is a rare disease with low malignant potential. The resection site and the peritoneal cavity are the most common sites of tumor relapse. Despite extensive peritoneal involvement, the presence of regional lymph nodes and hematogenous metastases is exceptional. We report four cases of appendiceal carcinoma metastatic to the right psoas muscle/aortoiliac region and hypothesize regarding the mechanisms of dissemination. We use our experience with this unusual condition to make recommendations regarding treatment. PMID- 12575803 TI - Giant mediastinal parathyroid adenoma with hypercalcemia. AB - Parathyroid disease is uncommon, but when found it is always of interest to the physician. Most patients with parathyroid disease also have hypercalcemia. Benign adenomatous disease is the most frequent surgical parathyroid lesion seen. Most of these lesions occur in the neck and are usually <2 cm in diameter. We report the case of a 50-year-old patient with familial neurofibromatosis, a serum calcium of 19 mg/dl, nephrocalcinosis, and renal failure. Evaluation revealed a large (5.7 x 4.5 x 2.9-cm) mediastinal adenoma. At resection, the tumor weighed 39.5 g. Symptomatology rapidly improved postoperatively. PMID- 12575804 TI - Choledochocele: the disputed origin. AB - Choledochocele is classified as a type III choledochal cyst according to Alonso Lej classification. Although it is anatomically related to the common bile duct, it does not share the strong premalignant potential of the more common types of choledochal cysts. In addition, duodenal mucosa has been found lining many of these cysts. These findings pose certain disputes about their possible origin, necessitating revision of the current inclusion of choledochocele with choledochal cysts. PMID- 12575805 TI - Validity and limits of intraoperative rapid parathyroid hormone assay in primary hyperparathyroidism treated by traditional and mini-invasive surgery. AB - The aim of this work was to analyze patients suffering from primary hyperparathyroidism (HPT) treated by traditional and miniaccess surgery, to demonstrate the validity and limits of intraoperative assay of PTH (iPTH). Between January 2000 and December 2001, at the Surgical Science Department of "La Sapienza" University of Rome, 29 patients affected by HPT, one of whom was a recidivist, underwent surgery for HPT and had a mean follow-up of 15 months (range, 6-24 months). The research showed that a decrease of >50% in iPTH values is indicative of resolution of clinical signs in 95% of cases. The use of iPTH in HPT for solitary adenoma in both classical and mini-invasive surgery yields a 100% success rate in terms of persistence, recidivism, and postoperative normocalcemia. iPTH is of assistance to the surgeon and allows treatment through mini-invasive access. In the case of solitary adenoma, iPTH is not only a biochemical histological examination but also a predictive test of normocalcemia. PMID- 12575806 TI - Reconstruction of the hepatic vein using a patch graft from the autologous pericardium. AB - We describe a patch graft technique using the autologous pericardium for reconstruction of the right hepatic vein after hepatectomy. A male patient underwent hepatectomy for removal of metastatic tumors from colon cancer. A 2.5 cm-diameter tumor, which was located in the antero-superior segment (S8) of the right lobe, invaded the right hepatic vein. A patch graft was obtained from the autologous pericardium. After clamping of the proximal and distal parts of the right hepatic vein invaded by the cancer, part of the anterior wall, measuring 2.0 x 1.5 cm, was resected. The patch fitted the defect in the right hepatic vein well and was sutured in place using running sutures of 6-0 Proline. Its patency was maintained 14 months after surgery. This technique can be applied for reconstruction after partial resection of a vein in this and other sites. PMID- 12575807 TI - Role of preoperative color doppler ultrasound scan in the evaluation of the risk of injury of major branches of the middle hepatic vein during laparoscopic cholecystectomy. AB - During laparoscopic cholecystectomy, the separation of the gallbladder from the liver bed may sometimes cause severe hemorrhages. One reason for severe hemorrhages may be injury to the major branches of the middle hepatic vein (MHV), which may be too close or adherent to the gallbladder. In our institutional experience of 798 laparoscopic cholecystectomies, no major hemorrhage from the gallbladder bed has been encountered. The aim of this prospective study was to investigate the relationship between the major branches of the MHV and the gallbladder bed in our patients. We measured the distance of the closest branches of the MHV from the gallbladder bed by color Doppler ultrasound scan. The mean and the median distances of the closest branch of the MHV to the gallbladder was found to be 17.4 +/- 6.2 mm and 17.7 mm, respectively (range, 6-29.1 mm). In conclusion, the distance of the closest branch of the MHV to the gallbladder bed in our patient population seems to allow for a safe laparoscopic cholecystectomy. PMID- 12575808 TI - Influence of flail chest on outcome among patients with severe thoracic cage trauma. AB - Flail chest is associated with a higher morbidity compared with multiple rib fractures, and it requires early intubation. This was a prospective comparative uncontrolled study at an academic level 1 trauma center. Twenty-two patients with flail chest (FLAIL) were compared with 90 patients with more than two rib fractures but no flail chest (RIBS) to determine differences in outcomes such as mortality, significant respiratory complications (pneumonia and adult respiratory distress syndrome), need for mechanical ventilation, and length of hospital stay. Stepwise logistic regression identified independent risk factors of poor outcome. Despite similar age and rates of lung contusion and extrathoracic injury, FLAIL patients had a higher need for mechanical ventilation (86% versus 42%, P < 0.01), higher incidence of significant respiratory complications (64% versus 26%, P < 0.01), and longer hospital stay (28 +/- 21 versus 17 +/- 19 days, P = 0.04) compared with RIBS patients. Flail chest and extrathoracic injuries were independent risk factors of significant respiratory complications. Of 11 FLAIL patients who were not intubated on arrival, eight required intubation within the next 24 hours, often while receiving diagnostic studies in poorly monitored hospital areas; two of these patients suffered morbidity directly related to the delay in intubation. Three patients without associated injuries were managed successfully without intubation. Flail chest is an independent marker of poor outcome among patients with thoracic cage trauma. The majority of patients with flail chest need mechanical ventilatory support and develop significant respiratory complications. In the presence of associated injuries, intubation is unavoidable and should be done under controlled conditions early after arrival to avoid morbidity related to sudden respiratory decompensation. PMID- 12575809 TI - Postsplenectomy immunodepression and its implications: an underestimated problem. AB - The most important sequelae of splenectomy is immunity depression. This study, conducted in three phases, was aimed at confirming this clinical condition. Data from our phase 1 study clearly show that patients undergoing splenectomy for trauma are in a critical condition because of a latent immunodeficiency shown by skin tests (ST) and in vitro evaluation of the aspecific immune activity. Because the in vitro study of the unspecific immunity that we used seems to be more expensive and complicated than ST, the aim of the phase 2 study was to compare the efficacy and the limits of the two assays (ST versus in vitro study) in detecting the immunodeficiency status of the splenectomized patient. The aim of the phase 3 study was to ascertain whether postsplenectomy immunodeficiency could be a consequence of an altered equilibrium between the lymphocyte subpopulations T helper (Th)1/Th2, evaluated by serum dosage of interferon-gamma and interleukin 4. PMID- 12575810 TI - Bronchoscopy and anesthesia for preschool-aged patients: a review of 228 cases. AB - Pediatric bronchoscopy is a complicated, high-risk procedure. From May 2000 to May 2002, we performed bronchoscopy in 228 preschool-aged children (newborn to 6 years old) under general anesthesia with a Storz-Hopkins rigid pediatric bronchoscope. The final diagnosis and complications during anesthesia were recorded. The most common findings from bronchoscopic examination were subglottic stenosis (or granuloma) (67, 29.4%), laryngomalacia (64, 28.1%), tracheal stenosis (or malacia) (29, 12.7%), pneumonia (23, 10.1%), and atelectasis (16, 7.0%). The main complications during anesthesia were arrhythmias, oxygen desaturation, and CO2 retention (high end-tidal CO2). The most serious complication was pneumothorax in one patient. Either endotracheal intubation or tracheostomy was required in 61% of the patients in this series to secure a patent airway after bronchoscopy. Bronchoscopy is necessary as a diagnostic and therapeutic tool for certain airway diseases or anomalies in pediatric patients. It requires cooperation between the endoscopist and anesthesiologist to insure the patient's safety. PMID- 12575811 TI - Peutz-Jeghers syndrome associated with intestinal intussusception: a case report. AB - A 12-year-old boy visited our hospital with complaints of severe upper abdominal pain and vomiting. Marked tenderness was present in the upper abdomen, and a movable hard tumor the size of a fist was felt there. Because abdominal computed tomography scans revealed invagination, an emergency laparotomy was performed. The small intestine was invaginated over an area measuring 30 cm in length. After the invaginated intestine was restored, a mass was felt in the small intestine, which was resected over an area measuring approximately 10 cm in length. The extracted mass was histopathologically found to be a hamartoma. The patient was diagnosed to have Peutz-Jeghers syndrome (PJS) based on the presence of pigmented spots detected on the oral cavity, lips, and toes of both feet. PJS is an autosomal dominant genetic disease associated with melanin pigment spots on the oral mucosa, lips, nasal alae, palm, and soles, as well as hamartomatous polyposis in the alimentary canal. Polyps are often a cause of invagination and ileus in affected patients. Gastrointestinal cancer, uterine cancer, and breast cancer are also seen in patients with PJS. A long-term follow-up is required to prevent invagination and ileus in children and cancer in adults. PMID- 12575812 TI - Heparinase enhances collateral vessel development in the ischemic limb. AB - Several angiogenic factors are bound to heparin and stored in the extracellular matrix of diverse tissues; thus, the controlled release of these factors can provide a novel approach of angiogenic stimulation to ischemic tissues. The purpose of this study was to test the hypothesis that heparinase, when administered in vivo, might enhance collateral vessel development in the ischemic limb by the controlled release of angiogenic factors such as basic fibroblast growth factor (bFGF). Eleven male New Zealand White rabbits underwent ligation and excision of the common and superficial femoral arteries in the left hindlimb. In the heparinase group (n = 6), 9.7 IU of heparinase (in 3 ml saline) was injected intramuscularly to the left thigh daily for 10 days, beginning 11 days after surgery. In the control group (n = 5), inactivated heparinase (also in 3 ml saline) was administered following the same experimental protocol. Calf systolic pressure was measured in both hindlimbs and was expressed as a ratio of left to right (L/R ratio) before injections (on day 10) and after injections (on days 20, 30, and 40). Vascularization was quantified by comparing the number of vessels along a line drawn across the mid-thigh on angiograms taken 4 seconds after contrast injection at day 40 when the study was terminated. The intramuscular injection of heparinase improves perfusion to the ischemic limb through the process of enhanced collateral vessel development. The angiogenic effect of heparinase on the ischemic tissue may result from release of endogenous angiogenic factors, such as bFGF. PMID- 12575813 TI - Splenectomy for children with thalassemia. AB - Twenty-three children with thalassemia (18 with beta-thalassemia major, 3 with Hb H disease, and 2 with thalassemia intermediate) had total splenectomy (nine beta thalassemia major patients and two thalassemia intermediate patients) or partial splenectomy (nine beta-thalassemia patients and three Hb H disease patients) as part of their management at our hospital. There were 10 males and 1 female in the total splenectomy group (mean age, 7.8 years; range, 4.5-12 years), and 4 males and 8 females in the partial splenectomy group (mean age, 6.9 years; range, 3-10 years). In all, the indication for splenectomy was hypersplenism. In the partial splenectomy group, two children with Hb H disease required no further blood transfusions. The transfusion requirements of the third patient with Hb H disease decreased from 15 to 11 transfusions per year (from 1.2 g/week Hb drop preoperatively to 0.7 g/week postoperatively), but subsequently his transfusion requirements increased as a result of an increase in the size of splenic remnant. He underwent total splenectomy 1.5 years post-partial splenectomy. For those with beta-thalassemia major who had partial splenectomy, there was a reduction in the number of blood transfusions from a preoperative mean of 15.2 transfusions per year (range, 11-22 transfusions per year) to a postoperative mean of 8.2 transfusions per year (range, 2-11 transfusions per year). Their Hb drop decreased from a preoperative mean of 1.6 g/week (range, 0.8-3.5 g/week) to a postoperative mean of 0.5 g/week (range, 0.2-0.75 g/week). Subsequently and as a result of increase in the size of splenic remnant, their transfusion requirements increased, but none of them to this point have required total splenectomy. Eleven children had total splenectomy. Their postsplenectomy transfusion requirements decreased from a preoperative mean of 17.8 transfusions per year (range, 12-23 transfusions per year) to a postoperative mean of 10 transfusions per year (range, 8-12 transfusions per year), and their Hb drop decreased from a preoperative mean of 1.8 g/week (range, 0.5-2.3 g/week) to a postoperative mean of 0.45 g/week (range, 0.3-0.65 g/week). In conclusion, total splenectomy is beneficial for children with thalassemia and hypersplenism because it reduces their transfusion requirements. Partial splenectomy may be beneficial for those with Hb H disease. However, for those with beta-thalassemia, partial splenectomy is beneficial in reducing their transfusion requirements only as a temporary measure, and it is recommended for children who are less than 5 years of age. PMID- 12575814 TI - Low dose discontinued FK506 treatment enhances peripheral nerve regeneration. AB - FK506 has been reported to enhance nerve regeneration in various rodent models. However, both the immunosuppressive properties and potential side effects may preclude the broad clinical use of FK506 to speed nerve regeneration. In this study, we examined the nerve regenerative property of a low, nonimmunosuppressive dose (0.5 mg/kg/day) of FK506. Two treatment schedules (continuous versus discontinuous) were compared. Evaluations were perforrmed at 1, 2, and 3 months. The gastrocnemius muscle mass was significantly higher in treated groups compared with the untreated group at 3 months, and the total number of fibers, percentage of neural tissue, fiber density, and fiber diameter were greater in treated groups than in the untreated group. However, at 3 months there was no difference in these parameters between groups in which FK506 was administered continuously and those in which the drug was stopped at 2 months. This study confirms that daily administration of low dose FK506 enhances peripheral nerve recovery after transection injury. PMID- 12575815 TI - Transjugular intrahepatic portosystemic shunt migration in patients undergoing liver transplantation. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is a useful procedure for patients with variceal bleeding and refractory ascites. Migration of TIPS can potentially complicate the subsequent transplant procedure. The aim of this study was to compare survival, operating time, and blood transfusion requirements in patients with migrated and nonmigrated TIPS undergoing liver transplantation. Of 152 patients, 21 received TIPS; stent migration was noted in seven patients-six distally and one proximally. Mean age of the patients was 54 +/- 11 years (range, 27-65 years), and there were 12 men and 9 women. The etiology of liver disease included the following: hepatitis C virus, six patients; cryptogenic cirrhosis, seven patients; alcoholic cirrhosis, four patients; primary biliary cirrhosis, three patients; and autoimmune hepatitis, one patient. The mean Child-Pugh Turcotte score was 10 +/- 2. Mean length of hospital stay for patients with migrated TIPS was 22.2 days and for nonmigrated TIPS was 23.5 days. Patient and graft survival (actual) was 81% in both groups with a mean follow-up of 27.9 months. Migration of TIPS is not rare, and in our study it did not affect survival, length of surgery, or blood transfusion requirements compared with patients in whom TIPS had not migrated. PMID- 12575816 TI - Excitatory amino acid neurotoxicity. PMID- 12575818 TI - Detrimental and beneficial effects of injury-induced inflammation and cytokine expression in the nervous system. AB - Lesions in the nervous system induce rapid activation of glial cells and under certain conditions additional recruitment of granulocytes, T-cells and monocytes/macrophages from the blood stream triggered by upregulation of cell adhesion molecules, chemokines and cytokines. Hematogenous cell infiltration is not restricted to infectious or autoimmune disorders of the nervous system, but also occurs in response to cerebral ischemia and traumatic lesions. Neuroinflammation can cause neuronal damage, but also confers neuroprotection. Granulocytes occlude vessels during reperfusion after transient focal ischemia, while the functional role of T-cells and macrophages in stroke development awaits further clarification. After focal cerebral ischemia neurotoxic mediators released by microglia such as the inducible nitric oxide synthase (leading to NO synthesis) and the cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) are upregulated prior to cellular inflammation in the evolving lesion and functionally contribute to secondary infarct growth as revealed by numerous pharmacological experiments and by use of transgenic animals. On the other hand, cytokine induction remote from ischemic lesions involves NMDA-mediated signalling pathways and confers neuroprotection. After nerve injury T cells can rescue CNS neurons. In the peripheral nervous system neuroinflammation is a prerequisite for successful regeneration that is impeded in the CNS. In conclusion, there is increasing evidence that neuroinflammation represents a double edged sword. The opposing neurotoxic and neuroprotective properties of neuroinflammation during CNS injury provide arich and currently unexplored set of research problems. PMID- 12575817 TI - Neuronal survival and cell death signaling pathways. AB - Neuronal viability is maintained through a complex interacting network of signaling pathways that can be perturbed in response to a multitude of cellular stresses. A shift in the balance of signaling pathways after stress or in response to pathology can have drastic consequences for the function or the fate of a neuron. There is significant evidence that acutely injured and degenerating neurons may die by an active mechanism of cell death. This process involves the activation of discrete signaling pathways that ultimately compromise mitochondrial structure, energy metabolism and nuclear integrity. In this review we examine recent evidence pertaining to the presence and activation of anti- and pro-cell death regulatory pathways in nervous system injury and degeneration. PMID- 12575819 TI - Cellular and molecular determinants of glial scar formation. PMID- 12575820 TI - Na+ channels and Ca2+ channels of the cell membrane as targets of neuroprotective substances. PMID- 12575821 TI - Intracellular Ca2+ handling. AB - Intracellular Ca2+ is regulated within three major compartments: the cytosol, the endoplasmic reticulum and mitochondria. This Chapter reviews the mechanisms involved in handling of Ca2+ within these compartments with reference to potential strategies for neuroprotection. In the cytosol, Ca2+ buffering has a major influence on Ca2+ signals. Cytosolic Ca(2+)-binding proteins such as CB28 participate in Ca2+ buffering and may have a role in resistance to neurotoxicity. In the endoplasmic reticulum, a number of proteins are involved in Ca2+ uptake, lumenal buffering or release, and these may be of value as potential targets for therapeutic intervention. Mitochondria are receiving increasing attention for their role in Ca2+ storage and signaling, and as key players in the processes leading to cell death following Ca2+ overload. An improved understanding of how Ca2+ is controlled within these intracellular compartments, and how these compartments interact, will be important for neuroprotective strategies. PMID- 12575822 TI - Neuroprotective activity of metabotropic glutamate receptor ligands. AB - Metabotropic glutamate receptors form a family of currently eight subtypes (mGluR1-8), subdivided into three groups (I-III). Activation of group-II (mGluR2 and -3) or group-III metabotropic glutamate receptors (mGluR4, -6, -7 and -8) has been established to be neuroprotective in vitro and in vivo. In contrast, group-I mGluRs (mGluR1 and -5) need to be antagonized in order to evoke protection. Initially, all neuroprotective mGluR ligands were analogues of L-glutamate. Those compounds were valuable to demonstrate protection in vitro, but showed limited applicability in animal models, particularly in chronic tests, due to low blood brain-barrier penetration. Recently, systemically active and more potent and selective ligands became available, e.g., the group-II mGluR agonists LY354740 and LY379268 or group-I antagonists like MPEP (mGluR5-selective) and BAY36-7620 (mGluR1-selective). This new generation of pharmacological agents allows a more stringent assessment of the role of individual mGluR-subtypes or groups of receptors in various nervous system disorders, including ischaemia-induced brain damage, traumatic brain injury, Huntington's and Parkinson's-like pathology or epilepsy. Moreover, the use of genetically modified animals (e.g., knock-out mice) is starting to shed light on specific functions of mGluR-subtypes in experimental neuropathologies. PMID- 12575823 TI - A role for glutamate transporters in neurodegenerative diseases. PMID- 12575824 TI - Purines and neuroprotection. AB - The activation of adenosine A1, A2 andA3 receptors can protect neurones against damage generated by mechanical or hypoxic/ischaemic insults as well as excitotoxins. A1 receptors are probably effective by suppressing transmitter release and producing neuronal hyperpolarisation. They are less likely to be of therapeutic importance due to the plethora of side effects resulting from A1 agonism, although the existence of receptor subtypes and recent synthetic chemistry efforts to increase ligand selectivity, may yet yield clinically viable compounds. Activation of A2A receptors can protect neurons, although there is much uncertainty as to whether agonists are acting centrally or via a peripheral mechanism such as altering blood flow or immune cell function. Selective antagonists at the A2A receptor, such as 4-(2-[7-amino-2-(2 furyl)(1,2,4)triazolo(2,3-a)(1,3,5)triazin-5-yl-amino]ethyl)phenol (ZM 241385) and 7-(2-phenylethyl)-5-amino-2-(2-furyl)-pyrazolo-[4,3e]-1,2,4-triazolo[1,5 c]pyrimidine (SCH 58261), can also protect against neuronal death produced by ischaemia or excitotoxicity. In addition, A2A receptor antagonists can reduce damage produced by combinations of subthreshold doses of the endogenous excitotoxin quinolinic acid and free radicals. Since the A2A receptors do not seem to be activated by normal endogenous levels of adenosine, their blockade should not generate significant side effects, so that A2A receptor antagonists appear to be promising candidates as new drugs for the prevention of neuronal damage. Adenosine A3 receptors have received less attention to date, but agonists are clearly able to afford protection against damage when administered chronically. Given the disappointing lack of success of NMDA receptor antagonists in human stroke patients, despite their early promise in animal models, it is possible that A2A receptor antagonists could have a far greater clinical utility. PMID- 12575825 TI - Heat shock proteins and neuroprotection. AB - In response to many metabolic disturbances and injuries including stroke, neurodegenerative disease, epilepsy and trauma, the cell mounts a stress response with induction of a variety of proteins, most notably the 70 kD heat shock protein (Hsp70). The possibility that stress proteins might be neuroprotective was suspected because Hsp70, in particular, was induced to high levels in brain regions that were relatively resistant to injury. Hsp70 expression was also correlated with the phenomenon of induced tolerance. With the availability of transgenic animals and gene transfer, has it become increasingly clear that such heat shock proteins do indeed protect cells from injury. Several reports have now shown that selective overexpression of Hsp70 leads to protection in several different models of nervous system injury. This review will cover these studies, along with potential mechanisms by which Hsp70 might mediate cellular protection. PMID- 12575826 TI - Neurotrophins. AB - Nerve growth factor was the first identified protein with anti-apoptotic activity on neurons. This prototypic neurotrophic factor, together with the three structurally and functionally related growth factors brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3) and neurotrophin-4/5 (NT4/5), forms the neurotrophin protein family. Target T cells for neurotrophins include many neurons affected by neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and peripheral polyneuropathies. In addition, the neurotrophins act on neurons affected by other neurological and psychiatric pathologies including ischemia, epilepsy, depression and eating disorders. Work with cell cultures and animal models provided solid support for the hypothesis that neurotrophins prevent neuronal death. While no evidence exists that a lack of neurotrophins underlies the etiology of any neurodegenerative disease, these studies have spurred on hopes that neurotrophins might be useful symptomatic-therapeutic agents. However first clinical trials led to variable results and severe side effects were observed. For future therapeutic use of the neurotrophins it is therefore crucial to expand our knowledge about their physiological functions as well as their pharmacokinetic properties. A major challenge is to develop methods for their application in effective doses and in a precisely timed and localized fashion. PMID- 12575827 TI - Fibroblast growth factors and neuroprotection. AB - Several members of the FGF family, in particular FGF2, are intimately involved in neuronal protection and repair after ischemic, metabolic or traumatic brain injury. Expression of Fgf2 mRNA and protein is strongly upregulated after neuronal damage, with glial cells as the predominant source. Given its survival promoting effects on cultured neurons, exogenous FGF2 was tested in several animal models of stroke and excitotoxic damage, in which it consistently proved protective against neuronal loss. FGF2 affords neuroprotection by interfering with a number of signaling pathways, including expression and gating of NMDA receptors, maintenance of Ca2+ homeostasis and regulation of ROS detoxifying enzymes. FGF2 prevents apoptosis by strengthening anti-apoptotic pathways and promotes neurogenesis in adult hippocampus after injury. The protective action of FGF2 has been linked to its augmenting effect on the lesion-induced upregulation of activin A, a member of the TGF-beta superfamily. Despite the well-documented benefits of FGF2 in animal models of stroke, there is currently no clinical development in stroke, after a phase II/III trial with FGF2 in acute stroke patients was discontinued because of an unfavorable risk-to-benefit ratio. As the molecular targets of FGF2 are going to be unraveled over the next years, new therapeutic strategies will hopefully emerge that enable us to influence the various protective mechanisms of FGF2 in a more specific fashion. PMID- 12575828 TI - TGF-betas and their roles in the regulation of neuron survival. AB - Transforming growth factor-betas (TGF-betas) are a still growing superfamily of cytokines with widespread distribution and diverse biological functions. They fall into several subfamilies including the TGF-betas 1, 2, and 3, the bone morphogenetic proteins (BMPs), the growth/differentiation factors (GDFs), activins and inhibins, and the members of the glial cell line-derived neurotrophic factor family. Following a brief description of their general roles and signaling in development, maintenance of homeostasis, and disease, we shall focus on their distribution in the CNS and their involvement in regulating neuron survival and death. PMID- 12575829 TI - Vascular endothelial growth factor. PMID- 12575831 TI - Invasive drug delivery. AB - The central nervous system is a very attractive target for new therapeutic strategies since many genes involved in neurological diseases are known and often only local low level gene expression is required. However, as the blood brain barrier on one hand prevents some therapeutic agents given systematically from exerting their activity in the CNS, it also provides an immune privileged environment. Neurosurgical technology meanwhile allows the access of nearly every single centre of the CNS and provides the surgical tool for direct gene delivery via minimal invasive surgical approaches to the brain. Successful therapy of the central nervous system requires new tools for delivery of therapeutics in vitro and in vivo (Fig. 1). The application of therapeutic proteins via pumps into the CSF was shown to be only of limited value since the protein mostly is not sufficiently transported within the tissue and the half life of proteins limits the therapeutic success. Direct gene delivery into the host cell has been a main strategy for years, and in the beginning the direct DNA delivery or encapsulation in liposomes or other artificial encapsulation have been applied with different success. For several years the most promising tools have been vectors based on viruses. Viruses are able to use the host cell machinery for protein synthesis, and some of them are able to stably insert into the host cell genome and provide long term transgene expression as long as the cell is alive. The increasing knowledge of viruses and their live cycle promoted the development of viral vectors that function like a shuttle to the cell, with a single round of infection either integrating or transiently expressing the transgene. Viral vectors have proven to be one of the most efficient and stable transgene shuttle into the cell and have gained increasing importance. The limitations of some viral vectors like the adenoviral vector and adeno-associated viral vector have been improved by new constructs like HIV-1 based lentiviral vectors. The immune response caused by expression of viral proteins, or the inability of some viral vectors like the retroviral vector to infect only dividing cells have been overcome by these new constructs. Lentiviral vectors allow an efficient and stable transgene expression over years in vivo without effecting transgene expression or immune response. In this Chapter we will describe synthetic vectors, give an overview of the most common viral vectors and focus our attention on lentiviral vectors, since we consider them to be the most efficient tool for gene delivery in the CNS. PMID- 12575830 TI - Blood-brain barrier drug targeting enables neuroprotection in brain ischemia following delayed intravenous administration of neurotrophins. AB - The blood-brain barrier (BBB) is the rate-limiting step in the translation of neurotrophin neuroscience into clinically effective neurotherapeutics. Since neurotrophins do not cross the BBB, these proteins cannot be used for neuroprotection following intravenous administration, and it is not feasible to administer these molecules by intra-cerebral injection in human stroke. The present studies describe the development of the chimeric peptide brain drug targeting technology and the use of brain-derived neurotrophic factor (BDNF) chimeric peptides in either global or regional brain ischemia. The BDNF chimeric peptide is formed by conjugation of BDNF to a monoclonal antibody (MAb) to the BBB transferrin receptor, and the MAb acts as a molecular Trojan Horse to ferry the BDNF across the BBB via transport on the endogenous BBB transferrin receptor. High degrees of neuroprotection in transient forebrain ischemia, permanent middle cerebral artery occlusion, or reversible middle cerebral artery occlusion are achieved with the delayed intravenous administration of BDNF chimeric peptides. In contrast, no neuroprotection is observed following the intravenous administration of unconjugated BDNF, because the neurotrophin does not cross the BBB in vivo. PMID- 12575832 TI - Ischemia and stroke. AB - Cell death following cerebral ischemia is mediated by a complex pathophysiologic interaction of different mechanisms. In this Chapter we will outline the basic principles as well as introduce in vitro and in vivo models of cerebral ischemia. Mechanistically, excitotoxicity, peri-infarct depolarization, inflammation and apoptosis seem to be the most relevant mediators of damage and are promising targets for neuroprotective strategies. PMID- 12575833 TI - Neuroprotective strategies in Alzheimer's disease. PMID- 12575834 TI - The pattern of cone pedicles and horizontal cells in the retina of the European anchovy, Engraulis encrasicolus L. (Engraulididae, Clupeiformes). AB - We have studied the architecture of polycone pedicles and their postsynaptic horizontal cells in the European anchovy using serial reconstruction by means of thin-sections from the cone outer segments to the outer plexiform layer. Within this layer there is a tesselated arrangement of pedicles in two strata, one made by the long-cone pedicles and one by those of the short-cones. The two pedicle types differ in the architecture of their synaptic sites. The long-cones possess two ribbon clusters corresponding to two bunches of subsynaptic dendrites, while the short-cones have only a single ribbon cluster innervated by a single group of dendrites. In addition, telodendria specifically connect the cone pedicles of neighbouring rows. Among the horizontal cells three classes arranged in separate levels can be distinguished. Serial reconstruction results suggest that first level horizontal cells contact long-cones preferentially. Among bipolar cells with an oval shaped dendritic field another type is found with a rectangular field of polycone-parallel dendritic combs. These findings indicate that the uncommon cone structures of engraulidids, which probably subserve polarization contrast vision, are correlated with a peculiar geometry of the outer plexiform layer. PMID- 12575835 TI - Skeletal muscle regeneration associated with the stroma reaction during tumor invasion in the rat tongue. AB - This study was aimed to demonstrate the regeneration of skeletal muscle fibers in the stroma reaction during tumor invasion, using the rat model of tongue carcinoma. By oral administration of 4-nitroquinoline N-oxide, squamous cell carcinoma (SCC) appeared in the epithelium, and deeply invaded the muscular layer, inducing the stroma reaction around the tumor. Regenerating muscle fibers, characterized by the immature profiles of sparse myofibrils, centrally disposed multi-nuclei, and abundant mitochondria, were extended from the surrounding normal muscles into the stroma. By immunohistochemistry, some of them expressed BF-45, a marker for an early stage of myodifferentiation, similar to the regenerating muscle fibers in the bupivacaine hydrochloride-induced injury. They were closely associated with the stromal components such as ED-1-positive macrophages, alpha-smooth muscle actin-positive myofibroblasts, and factor VIII related antigen-positive vascular endothelial cells, suggesting the roles of their interactions in muscle regeneration. Immature muscle fibers were usually devoid of acetylcholinesterase-positive endplates on them, but some were reinnervated by the terminal axons. The present results indicate that skeletal muscle regeneration is induced in association with the stroma reaction during SCC invasion in the tongue. PMID- 12575836 TI - Localization of peroxidase activity in blood mononuclear phagocytes in pacu fish (Piaractus mesopotamicus). AB - The localization of peroxidase activity in different cell regions is used as a criterion for classifying the stage of maturity of mammalian mononuclear phagocytes, with a positive peroxidase reaction indicating the presence of monoblasts, promonocytes, monocytes, and macrophages. Peroxidase activity was observed ultrastructurally in the circulating blood of pacu fish (Piaractus mesopotamicus), identifying monoblasts, promonocytes, monocytes, and macrophages. These observations suggest that differentiation of mononuclear phagocytes occurs in the blood circulation of fish, whereas in mammals, monoblasts and promonocytes are detected in bone marrow, with only monocytes detected in circulating blood and differentiation into macrophages occurring in other body compartments. PMID- 12575837 TI - Effects of nicotine on intestinal and respiratory epithelium. AB - The effects of nicotine on intestinal and tracheal mucosa and epithelial cells were studied in vivo and in vitro. Rats received 108 microM nicotine in their drinking water for 10 days. Intestine and trachea were removed and prepared for X ray microanalysis, transmission electron microscopy and immunocytochemistry. X ray microanalysis of freeze-dried cryosections of intestine and trachea showed an increase in Na and Cl, and a decrease in K in the lamina propria of the intestine and the epithelial cells and the submucosal compartment of the trachea. Analysis of frozen-hydrated trachea, in order to determine ionic changes in the airway surface liquid (ASL), indicated that the Na and Cl content in the ASL decreased after nicotine treatment. Immunocytochemistry showed upregulation of ICAM-1 in the submucosal connective tissue of the trachea and the lamina propria of the intestine. Transmission electron microscopy showed a somewhat increased number of eosinophils in the lamina propria of nicotine-treated rats, increased edema in the submucosal connective tissue, and a somewhat increased number of damaged basal cells in the trachea of nicotine-treated rats, compared to the controls. These data indicate that nicotine may evoke an inflammatory reaction, in particular in the trachea, that could cause cell damage and with that changes in the ionic relations of the epithelial cells. The in vitro experiments showed that nicotine could directly affect ion transport by inhibiting cAMP-stimulated (but not ATP-stimulated) chloride efflux from cultured respiratory epithelial cells. This indicates that apart from indirect effects via inflammation, nicotine can directly affect the ionic homeostasis of the cells and the composition of the airway-surface liquid. PMID- 12575838 TI - Morphological and ultrastructural study of the thrombocytes and leukocyte granulocytes of Salminus maxillosus (Characiformes, Characidae). AB - The thrombocytes and leukocytes of 'dourado' fish Salminus maxilosus peripheral blood were studied by means of light microscopy and transmission electron microscopy techniques. In respect to the morphological and ultrastructural aspects of thrombocytes and granulocytes, the cellular elements that appear with high frequency in the light microscopy preparations stained by the Rosenfeld method, three different types of granulocytes could be differentiated at the electron microscope, based on the ultrastructural aspect of the cytoplasmic elements: type I granulocyte that possibly corresponds to the type I neutrophil described under light microscopy; type II granulocyte is possibly the cell identified as type II neutrophil by the morphological analysis; type III granulocyte is equivalent to the eosinophils described in the samples stained by Rosenfeld methods. PMID- 12575839 TI - Morphometric evaluation of muscle fiber types in different skeletal muscles of rats. AB - The purpose of the present work was to study some morphological differences of similar muscle fiber types--classified by ATPase reactions in different muscles of rats. Morphological parameters were used as stereological techniques at light and electron microscopic level. There was a great variation in the diameter of each muscle fiber type of different muscles. The smallest diameter of type 1 myofibers of the soleus was greater than the diameter of type 1 myofibers of other muscles. The diameter of type 1 myofibers of the soleus and of the lateral part of the gastrocnemius was almost twice the diameter of type 1 myofibers of sternocleidomastoid. The lateral and medial parts of gastrocnemius had the largest 2A and 2B muscle fibers. As a whole, among the studied muscles, myofibers of postural muscles of the posterior parts of the posterior limbs had the greatest diameter. Stereological analysis at electron microscopic level revealed that there were differences in the volume density of mitochondria in the different muscles. The quantity of mitochondria was greater in the diaphragm than in the gastrocnemius, soleus and sternocleidomastoid muscles. Our results suggested that the diameter of muscle fibers is more related to the resistance the muscle is submitted than to the continuous necessity of contraction. However, the quantity of mitochondria of oxidative fibers of the diaphragm would be related to continuous necessity of contraction and high oxidative necessity of this muscle. PMID- 12575840 TI - Lack of amplification phenomena of ribosomal RNA genes in sea urchin. AB - Careful quantitative analyses by spot hybridization to homologous probes demonstrate that no rDNA amplification occurs during Paracentrotus lividus oogenesis. The same approach was used to measure the copy number of the genes involved in ribosome biogenesis. Surprisingly, differently from the organisms in which the lack of rDNA amplification phenomena was observed, a very low number of constitutive rDNA repeats was found in this organism. PMID- 12575841 TI - Light and electron microscopy oogenesis in matrinxa, Brycon cephalus. AB - Oocyte development has been divided into 5 distinct stages in the reared matrinxa, Brycon cephalus, based on morphological criteria by light and transmission electron microscopy: I) chromatin-nucleolus; II) perinucleolar; III) cortical alveoli; IV) vitellogenesis; V) final maturation. In stages I and II (primary growth), oocytes reside in nests close to other oocytes (chromatin nucleolus phase) and then within a definitive follicle (perinucleolar phase) where they greatly increase in size (the Balbiani vitelline body is the main cytoplasmic component in these latter oocytes), respectively. In stage III (cortical alveolus phase) oocytes are distinguished by the appearance of variably sized cortical alveoli and the number of these structures increases steadily towards hydration. The vitelline envelope becomes prominent. In the process of vitellogenesis (stage IV) one major accumulation of yolk proteins occurs in oocytes. In stage V (final maturation), oocyte increase slightly in size. Follicle cells go through a primordial stage and later change to a squamous and to a cubical shape. The chorion grows to a tripartite structure: an outer thin porous layer, an intermediate homogenous layer and an inner thick helicoidal layer. The ovulation of females matrinxa, required hormonal stimulation and this occurred 6 and 8 h after the second application. PMID- 12575842 TI - Putative inhibitory collicular boutons contact large neurons and their dendrites in the dorsal cochlear nucleus of the rat. AB - Within the circuits of the acoustic nuclei, the inferior colliculus sends descending (collicular) terminals to control with a feedback mechanism, part of the activity of the dorsal cochlear nucleus (DCN). It is not known whether this descending projection is prevalently excitatory or inhibitory. Using the neuronal tracer Wheat Germ Agglutinin conjugated to Horse Radish Peroxidase (WGA-HRP) the connections between the inferior colliculus and the DCN of the rat have been investigated. By far most retrograde labelled large neurons were glycine and GABA negative (pyramidal and giant neurons) and rare medium-size cells were glycine positive. The ultrastructural immunocytochemical analysis for glycine and GABA shows that mainly large, excitatory, neurons innervate the inferior colliculus. Rare medium-size glycine-positive cells with intermediate characteristics between pyramidal and cartwheel cells, seem also to project to the colliculus. Few WGA HRP labelled boutons contact the large cells or their dendrites, have symmetric pre- and post-synaptic thickenings, contain pleomorphic and/or flat vesicles, and are labelled for GABA or glycine. Since no GABA labelled cells in both the dorsal and ventral cochlear nucleus were retrograde labelled from the colliculus, the source of these intrinsic anterograde labelled boutons must be external to the cochlear nucleus. GABA positive neurons are both present in the inferior colliculus (injected with the tracer) and superior olivary complex (not injected with the tracer). This suggests that the double labelled boutons (WGA-HRP and GABA) are inhibitory GABA-ergic collicular terminals contacting the excitatory neurons of the DCN. Other few boutons or mossy fibers containing round vesicles and immunonegative for both glycine and GABA, were also seen contacting the large neurons and their dendrites in the DCN. As the round vesicles boutons may be derived from other retrograde cells of the cochlear nucleus (pyramidal and stellate cells) and those glycine positive from the glycinergic neurons in paraolivary nuclei, it is more likely that only the WGA-HRP and GABA labelled boutons are true collicular terminals. PMID- 12575843 TI - Kidney structural and ultrastructural pathological changes induced by uracoan rattlesnake (Crotalus vegrandis Klauber 1941) venom. AB - Acute renal insufficiency related to acute tubular necrosis is the most important complication caused by crotalid bite. For structural and ultrastructural studies of renal tissue, mice injected with crude venom or C. vegrandis haemorrhagic fraction, and controls were tested. Light microscopy analysis of kidneys at 24 h after injection of crude venom showed only moderate alterations such as tubular epithelia microvacuolisation. After 120 h marked glomerular and tubular capillaries congestion and interstitial oedema were observed. At 24 h after Uracoina-1 i.p. injection, intense glomerular and peritubular capillaries congestion was observed. Electron microscopic analysis of kidneys 24 h after i.p. injection of crude venom showed, capillary endothelial cell debris and pleomorphic mitochondria. Loss of interdigitations regularity, abundant dense bodies and light widening of the basal membrane were observed. Autophagic vacuoles were present as well as endothelia unfolding to the lumen and altered forms of podocytes. At 48 h, augmented endothelia without fenestrae formation with sequestration of low optical density debris inside the protrusions were noticed. At 120 h, capillary residues with loss of the endothelium were present and the basal membrane was widened. At 15 days, the number of vesicles and vacuoles in the tubules was increased and only few interdigitations were noticed. Autophagic vacuoles and mitochondrial matrix low electron density were observed. At 120 h after injection of crude venom, vascular damage with loss of capillary cell structures and collagen fibres were observed. At 24 h of haemorrhagic fraction injection, presence of autophagic vacuoles and myelinic figures were noticed. PMID- 12575844 TI - Trends and developments in the pharmacological treatment of psoriasis. PMID- 12575845 TI - Recombinant human erythropoietin influences revascularization and healing in a rat model of random ischaemic flaps. AB - In order to ascertain whether erythropoietin plays a role in early and late repair processes following ischaemic skin flap injury, a dorsal, caudally based skin flap was created in rats. The rats were successively divided into four groups. Group 1 was not treated. The other groups were treated with a subcutaneous administration of 0.9% NaCl saline solution (group 2), a subcutaneous administration of vehicle (group 3) or a subcutaneous administration of 300 IU/kg/day of recombinant human erythropoietin (group 4). We evaluated the possible relationships between neutrophil accumulation, myeloperoxidase activity and content in flap tissue, flap survival, flap temperature (using telethermography) and flap revascularization (using videocapillaroscopy). Necrosis in the flap was significantly less extensive in group 4 than in groups 1, 2 and 3. A significant increase in neutrophil infiltration occurred between the 1st and 24th hour in these groups, but this was not observed in group 4. These findings were confirmed by biochemical data of myeloperoxidase activity and malonyldialdehyde content. Between the 1st and 7th days, we recorded an increase of about 20% in flap temperature in groups 1, 2 and 3, whereas no significant variation was observed in group 4. On the 7th day, videocapillaroscopic findings showed an increase in the mean vascularization index in group 4. Our findings suggest that recombinant human erythropoietin administration can improve the wound healing process, in both early and late stages of injury, by reducing inflammatory response, increasing the density of capillaries in ischaemic flaps and allowing earlier repair of a damaged area. PMID- 12575846 TI - Sensitizing capacity of two monomeric aldehyde components in p-tert-butylphenol formaldehyde resin. AB - Contact allergy to p-tert-butylphenol-formaldehyde resin is not rare. This resin consists of a large number of substances, most of which are unknown. For diagnostic and preventive reasons, the chemical identity of the sensitizers should be known as well as their sensitizing capacities, cross-reaction patterns and presence in the environment. The aim of this study was to investigate the sensitizing capacities and cross-reaction patterns for 5-tert-butyl-2-hydroxy-3 hydroxymethyl-benzaldehyde and 5-tert-butyl-2-hydroxy-benzaldehyde in the guinea pig maximization test. 2,6-Dimethylol p-tert-butylphenol, p-tert-butylcate chol, 2-methylol p-tert-butylphenol, p-tert-butylphenol, 4-tert-butyl-2-(5-tert-butyl-2 hydroxy-3-hydroxymethyl-benzyloxymethyl)-6-hydroxymethyl-phenol and 4-tert-butyl 2-(5-tert-butyl-2-hydroxy-benzyloxymethyl)-phenol were used as potential cross reacting substances. 5-tert-Butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde was shown to be a sensitizer (p = 0.041). In animals induced with this compound no cross-reactions to the putative cross-reacting substances were seen. In contrast, 5-tert-butyl-2-hydroxy-benzaldehyde failed to induce sensitization and no cross reactions were detected. PMID- 12575847 TI - Bcl-2-related proteins, alpha-smooth muscle actin and amyloid deposits in aggressive and non-aggressive basal cell carcinomas. AB - Aberrant expression of bcl-gene products has been implicated in the development of non-melanoma skin cancers. Recently, altered expression of alpha-smooth muscle actin has been proposed as predictive of tumour behaviour in basal cell carcinomas. The purpose of this study was to compare the aggressive and non aggressive basal cell carcinomas in terms of bcl-gene products and alpha-smooth muscle actin expression. Fifty excisional biopsy samples were studied by immunohistochemical technique for the differential expressions of bcl-2, bax, bcl x and alpha-smooth muscle actin. Bcl-2, bcl-x and bax were expressed in 34 (68%), 38 (76%) and 41(82%) specimens, respectively. Immunoreactivity for alpha-smooth muscle actin was noted both in tumour nests (64%) and within the stroma (54%). There was a significant difference between aggressive and non-aggressive basal cell carcinomas in terms of bcl-2 and stromal alpha-smooth muscle actin immunoreactivity. Non-aggressive basal cell carcinomas display a concordant expression of bcl-family proteins, whereas aggressive tumours reveal a discordant pattern. An increased expression of stromal alpha-smooth muscle actin with a concomitant decrease or loss of bcl-2 expression may be highly suggestive of aggressiveness in basal cell carcinoma. PMID- 12575848 TI - Low frequency of beta-catenin gene mutations in pilomatricoma. AB - We investigated beta-catenin and adenomatous polyposis coli (APC) gene abnormalities in human pilomatricoma, in which a high incidence of beta-catenin gene mutations has been reported. Nucleated tumour cells were microdissected from 20 paraffin-embedded pilomatricomas. Exon 3 of the beta-catenin gene was amplified using polymerase chain reaction and sequencing analysis was performed. Immunostaining for beta-catenin and lymphoid-enhancer factor-1 was performed using the avidin-biotin-peroxidase method. Dinucleotide repeat markers D5S409 and D5S299 were used for polymerase chain reaction-based microsatellite analysis of the APC gene. The mutation cluster region of the APC gene was amplified using polymerase chain reaction and sequenced. Sequencing analysis revealed beta catenin gene mutations in 30%. All studied samples showed nuclear lymphoid enhancer factor-1 and cytoplasmic/nuclear beta-catenin expression. Loss of heterozygosity was observed in the APC gene, but no mutations in the mutation cluster region were found in seven tumours without beta-catenin mutations. The frequency of beta-catenin gene mutations was remarkably low, thus suggesting (i) the presence of mutations in other than exon 3 of the beta-catenin gene, (ii) a possible role of APC gene abnormalities, or (iii) involvement of other components of the Wingless-type MMTV integration site family pathway. PMID- 12575849 TI - Frequency of patch-test positivity in patients with psoriasis: a prospective controlled study. AB - Current information on the incidence of patch-test positivity and the spectrum of allergens in psoriatics is conflicting. We compared the rates of patch-test positivity to common allergens and topical medicaments in 200 patients suffering from chronic plaque psoriasis (group I) with 51 patients with other non-allergic skin complaints (group II) and 54 patients suspected of having allergic contact dermatitis (group III). Positive patch-test results to one or more allergens were detected in 21.6% of patients in group I, 23.5% in group II and 50.0% in group III. Psoriatics with > or = 5 years old disease had a higher rate of patch-test positivity than those with shorter disease duration (p<0.01). The site of lesions showed no correlation with patch-test positivity. The commonest allergens showing positivity in group I were dithranol (6.5%), nickel (6%), fragrance mix (5%), neomycin (2%) and nitrofurazone (2%). In spite of the comparable rates of patch test positivity in psoriatics and general dermatology outpatients, the predominance of sensitivity to topical medicaments and fragrance in the former group was striking. A separate psoriasis series focusing on topical agents may give more accurate information on this subject. PMID- 12575850 TI - Heliotherapy in atopic dermatitis: a prospective study on climatotherapy using the SCORAD index. AB - The aim of this study was to investigate the suitability and effectiveness of heliotherapy in moderate or severe atopic dermatitis in adults in the Canary Islands. A total of 216 patients participated on 6 different 2- or 3-week heliotherapy trips. Using the Severity Scoring of Atopic Dermatitis Index (SCORAD), the severity of atopic dermatitis was assessed prior to the start of heliotherapy, after 2 weeks and then 3 months after the end of heliotherapy. A quality-of-life questionnaire was later mailed to all participants. The mean SCORAD index was reduced by 70% after 2 weeks of heliotherapy and was still 45% lower 3 months after therapy (P<0.0001). At 3 months, the use of topical steroids was still significantly reduced (P<0.0001), whereas there was no significant (P=0.1166) change in the consumption of emollients. The quality of life of patients was improved and their self-treatment and working capacity was increased. As the longer 3-week period provided no significant additional advantage over a 2-week period, 2 weeks of heliotherapy can be considered optimal. In conclusion, heliotherapy is an effective and valuable therapeutic adjunct, especially in adults with severe atopic dermatitis. PMID- 12575851 TI - IgA pemphigus: the first two Scandinavian cases. AB - IgA pemphigus is a rare neutrophilic acantholytic skin disorder with only approximately 70 cases reported in the indexed literature to date. Here we describe two patients with IgA pemphigus (subcorneal pustular dermatosis type and intraepithelial neutrophilic type) that to our knowledge are the first Scandinavian patients with this disease. Initially, both patients were misdiagnosed as subcorneal pustular dermatosis of Sneddon and Wilkinson and only subsequent careful immunofluorescence studies (in one case with confocal laser scanning microscopy) led to the correct diagnosis. Apart from the expected IgA depositions on epidermal cell surfaces, both patients demonstrated some degree of intercellular IgG-specific immunofluorescence. No circulating IgA autoantibodies were detected. One patient was treated with the standard regime comprising dapsone and prednisolone, whereas in the other case a novel methotrexate and prednisolone combination treatment showed marked clinical efficacy. PMID- 12575852 TI - Anorectal involvement is frequent in limited systemic sclerosis. AB - The gastrointestinal tract, particularly the oesophagus, is affected in about half of all patients with systemic sclerosis. Only a few studies so far have dealt with the anorectal tract. We studied the anal function using anorectal manometry in 12 patients with limited systemic sclerosis. We also studied the oesophageal function. For the oesophagus, we measured the difference between intragastric and oesophageal pressure, while for the anorectal tract we investigated the maximum resting pressure, the maximum voluntary squeeze effort and the rectoanal inhibitory reflex. Maximum resting pressure and maximum voluntary squeeze effort were found to be decreased in all patients. The rectoanal inhibitory reflex was abnormal in four patients. Statistical analysis showed a significant correlation between maximum resting pressure and maximum voluntary squeeze effort. No correlation was found between oesophageal and anorectal involvement. Anorectal dysfunction is common in patients with limited systemic sclerosis. We suggest that these patients should have an evaluation of their anorectal function including anorectal manometry. PMID- 12575853 TI - Two men with toxic shock syndrome presenting with targetoid and spotty skin rashes. AB - Two previously healthy men who presented with hypotension, constitutional symptoms, and targetoid and discrete spotty erythematous plaques were diagnosed with toxic shock syndrome based on histopathological findings. Specifically, their biopsies revealed necrotic keratinocytes, neutrophils in the epidermis, and neutrophils surrounding dilated superficial vessels. In one case, the diagnosis of toxic shock syndrome was confirmed with rising titers to toxic shock syndrome toxin-1. Both patients recovered with supportive care and clindamycin administration. We suggest that patients with fever, hypotension, constitutional symptoms and rash should be started on clindamycin and have a skin biopsy as part of their initial evaluation. An understanding that toxic shock syndrome can strike anyone has manifold dermatological manifestations and defined histopathological findings is important for its early diagnosis and effective treatment. PMID- 12575854 TI - Superficial mucoceles and lichenoid graft versus host disease: report of three cases. AB - Superficial mucoceles are subepithelial extravasations of sialomucin that occur at the epithelial-connective tissue interface and are directly related to minor salivary glands. They have been described in association with oral lichen planus and, exceptionally, with chronic graft versus host disease. Three patients who underwent an allogeneic bone marrow transplantation for a chronic myelogenous leukaemia presented multiple superficial mucoceles and an oral lichenoid graft versus host disease. PMID- 12575855 TI - A variant of junctional naevus of epithelioid and spindle cell type rich in melanophages. AB - We describe a form of junctional melanocytic neoplasm with a massive production of melanin accumulated in the dermis. The pigment is stored in macrophages, which are by far the most numerous cellular component of the lesion. Another peculiar aspect is the occasional presence of a few melanocytes scattered in a pagetoid pattern above the dermo-epidermal junction in the spinous layer. The histological picture of this lesion is similar to a form of "tumoral melanosis" induced by a regressed malignant melanoma. The lesion had a worrisome clinical picture, its dark colour constituting a clinico-pathological diagnostic problem. The main clinical clues to the benign nature of this entity are the small lateral diameter, the uniform distribution of the pigment and the stability of the lesion over time; moreover, the patients are alive and well after a considerable length of time. Although a regressed dysplastic or malignant lesion cannot be totally excluded from a scientific point of view, we conclude that there is no sound morphological or clinical evidence that the lesion is other than biologically benign. The lesion is most likely another peculiar variant of epithelioid and spindle cell naevus. PMID- 12575856 TI - Non-invasive monitoring of compression therapy: a report of three cases with venous insufficiency. PMID- 12575857 TI - Atypical autoimmune blistering dermatosis associated with Sjogren's syndrome. PMID- 12575858 TI - Efficacy evaluation of an oil-in-water emulsion (Dermoflan) in atopic dermatitis. PMID- 12575859 TI - Prevalence of onychomycosis in Iceland: a population-based study. PMID- 12575860 TI - Leishmaniasis of the lip. PMID- 12575861 TI - Acute infantile haemorrhagic oedema: measles vaccination as possible triggering factor. PMID- 12575862 TI - Interstitial and granulomatous drug reaction presenting as erythema nodosum-like lesions. PMID- 12575863 TI - Successful treatment of vulvar intraepithelial neoplasia with topical imiquimod 5% cream in a lung transplanted patient. PMID- 12575864 TI - Successful treatment of basal cell carcinoma on the face with imiquimod 5% cream. PMID- 12575865 TI - Erythema ab igne caused by frequent hot bathing. PMID- 12575866 TI - An unusual manifestation of linear atrophoderma of Moulin. PMID- 12575867 TI - Perianal tuberculosis: a rare case of skin ulceration? PMID- 12575868 TI - Exacerbation of pemphigus foliaceus after tetanus vaccination accompanied by synthesis of auto-antibodies against paraneoplastic pemphigus antigens. PMID- 12575869 TI - Onychomycosis in children: treatment results of forty-seven patients. PMID- 12575870 TI - The Laboratory of Neuro Imaging: what it is, why it is, and how it came to be. PMID- 12575871 TI - Analysis of vasculature for liver surgical planning. AB - For liver surgical planning, the structure and morphology of the hepatic vessels and their relationship to tumors are of major interest. To achieve a fast and robust assistance with optimal quantitative and visual information, we present methods for a geometrical and structural analysis of vessel systems. Starting from the raw image data a sequence of image processing steps has to be carried out until a three-dimensional representation of the relevant anatomic and pathologic structures is generated. Based on computed tomography (CT) scans, the following steps are performed. 1) The volume data is preprocessed and the vessels are segmented. 2) The skeleton of the vessels is determined and transformed into a graph enabling a geometrical and structural shape analysis. Using this information the different intrahepatic vessel systems are identified automatically. 3) Based on the structural analysis of the branches of the portal vein, their vascular territories are approximated with different methods. These methods are compared and validated anatomically by means of corrosion casts of human livers. 4) Vessels are visualized with graphics primitives fitted to the skeleton to provide smooth visualizations without aliasing artifacts. The image analysis techniques have been evaluated in the clinical environment and have been used in more than 170 cases so far to plan interventions and transplantations. PMID- 12575873 TI - Longitudinal aliasing in multislice helical computed tomography: sampling and cone-beam effects. AB - In this study, we examine longitudinal aliasing properties in multislice helical computed tomography (CT) volumes reconstructed under the multiple parallel fanbeam approximation by use of a 180LI-type algorithm. We focus on the differences between the multislice case and the single-slice case, which has been studied previously. Specifically, we examine longitudinal aliasing properties in four-slice scanners for helical pitches 3 and 6, which are sometimes called "preferred" in four-slice helical CT, because it is believed that the effective longitudinal sampling intervals at these pitches are equivalent to those in single-slice helical CT operating at pitches 1 and 2, respectively. While these equivalences have been supported by comparative studies of slice-sensitivity profiles in single- and multislice helical CT, artifacts have been observed in pitch-3 and pitch-6 multislice images that were not evident in their purported single-slice counterparts. We attribute these differences to aliasing arising in the multislice reconstructions that is not present in the single-slice counterparts. We find that the aliasing has two principal origins: sampling effects similar to those in the single-slice case and cone-beam effects. The difference between the multislice, pitch-3 and single-slice, pitch-1 results is attributed to the small cone angle in multislice helical CT, which introduces inconsistencies among the measurements of different detector rows. The difference between multislice, pitch-6 and single-slice, pitch-2 results is attributed to a combination of the cone angle and genuine differences in sampling patterns. It is argued, however, that the lack of strict equivalence with single-slice counterparts does not necessarily undermine the claim that pitches 3 and 6 are "preferred" relative to other pitches in multislice helical CT. PMID- 12575872 TI - Mathematical generation of normal data for evaluating myocardial perfusion studies. AB - In this paper, we present a new mathematical method that synthesizes normal data sets for quantification of regional myocardium perfusion. In clinical practice, regional myocardial perfusion is often measured with a gamma camera and quantified via circumferential profile analysis. Normal reference profile data is used to increase the accuracy of the clinical interpretations. Our goal is to create reference data from an existing set of archived studies. An iterative mathematical method, based on two statistical hypotheses, was used to generate the study set instead of collecting normal examinations from a healthy population. Clinical validation is based on interpretations by six independent observers. Results of evaluation with synthesized normal data and its validation are presented. PMID- 12575874 TI - Automatic segmentation of echocardiographic sequences by active appearance motion models. AB - A novel extension of active appearance models (AAMs) for automated border detection in echocardiographic image sequences is reported. The active appearance motion model (AAMM) technique allows fully automated robust and time-continuous delineation of left ventricular (LV) endocardial contours over the full heart cycle with good results. Nonlinear intensity normalization was developed and employed to accommodate ultrasound-specific intensity distributions. The method was trained and tested on 16-frame phase-normalized transthoracic four-chamber sequences of 129 unselected infarct patients, split randomly into a training set (n = 65) and a test set (n = 64). Borders were compared to expert drawn endocardial contours. On the test set, fully automated AAMM performed well in 97% of the cases (average distance between manual and automatic landmark points was 3.3 mm, comparable to human interobserver variabilities). The ultrasound-specific intensity normalization proved to be of great value for good results in echocardiograms. The AAMM was significantly more accurate than an equivalent set of two-dimensional AAMs. PMID- 12575875 TI - Rapid elastic image registration for 3-D ultrasound. AB - A Subvolume-based algorithm for elastic Ultrasound REgistration (SURE) was developed and evaluated. Designed primarily to improve spatial resolution in three-dimensional compound imaging, the algorithm registers individual image volumes nonlinearly before combination into compound volumes. SURE works in one or two stages, optionally using MIAMI Fuse software first to determine a global affine registration before iteratively dividing the volume into subvolumes and computing local rigid registrations in the second stage. Connectivity of the entire volume is ensured by global interpolation using thin-plate splines after each iteration. The performance of SURE was quantified in 20 synthetically deformed in vivo ultrasound volumes, and in two phantom scans, one of which was distorted at acquisition by placing an aberrating layer in the sound path. The aberrating layer was designed to induce beam aberrations reported for the female breast. Synthetic deformations of 1.5-2.5 mm were reduced by over 85% when SURE was applied to register the distorted image volumes with the original ones. Registration times were below 5 min on a 500-MHz CPU for an average data set size of 13 MB. In the aberrated phantom scans, SURE reduced the average deformation between the two volumes from 1.01 to 0.30 mm. This was a statistically significant (P = 0.01) improvement over rigid and affine registration transformations, which produced reductions to 0.59 and 0.50 mm, respectively. PMID- 12575876 TI - Reconstruction in diffraction ultrasound tomography using nonuniform FFT. AB - We show an iterative reconstruction framework for diffraction ultrasound tomography. The use of broad-band illumination allows significant reduction of the number of projections compared to straight ray tomography. The proposed algorithm makes use of forward nonuniform fast Fourier transform (NUFFT) for iterative Fourier inversion. Incorporation of total variation regularization allows the reduction of noise and Gibbs phenomena while preserving the edges. The complexity of the NUFFT-based reconstruction is comparable to the frequency domain interpolation (gridding) algorithm, whereas the reconstruction accuracy (in sense of the L2 and the L(infinity) norm) is better. PMID- 12575877 TI - Functional MRI activity characterization using response time shift estimates from curve evolution. AB - Characterizing the response of the brain to a stimulus based on functional magnetic resonance imaging data is a major challenge due to the fact that the response time delay of the brain may be different from one stimulus phase to the next and from pixel to pixel. To enhance detectability, this work introduces the use of a curve evolution approach that provides separate estimates of the response time shifts at each phase of the stimulus on a pixel-by-pixel basis. The approach relies on a parsimonious but simple model that is nonlinear in the time shifts of the response relative to the stimulus and linear in the gains. To effectively use the response time shift estimates in a subspace detection framework, we implement a robust hypothesis test based on a Laplacian noise model. The algorithm provides a pixel-by-pixel functional characterization of the brain's response. The results based on experimental data show that response time shift estimates, when properly implemented, enhance detectability without sacrificing robustness. PMID- 12575878 TI - Generalization of median root prior reconstruction. AB - Penalized iterative algorithms for image reconstruction in emission tomography contain conditions on which kind of images are accepted as solutions. The penalty term has commonly been a function of pairwise pixel differences in the activity in a local neighborhood, such that smooth images are favored. Attempts to ensure better edge and detail preservation involve difficult tailoring of parameter values or the penalty function itself. The previously introduced median root prior (MRP) favors locally monotonic images. MRP preserves sharp edges while reducing locally nonmonotonic noise at the same time. Quantitative properties of MRP are good, because differences in the neighboring pixel values are not penalized as such. The median is used as an estimate for a penalty reference, against which the pixel value is compared when setting the penalty. In order to generalize the class of MRP-type of priors, the standard median was replaced by other order statistic operations, the L and finite-impluse-response median hybrid (FMH) filters. They allow for smoother appearance as they apply linear weighting together with robust nonlinear operations. The images reconstructed using the new MRP-L and MRP-FMH priors are visually more conventional. Good quantitative properties of MRP are not significantly altered by the new priors. PMID- 12575879 TI - HAMMER: hierarchical attribute matching mechanism for elastic registration. AB - A new approach is presented for elastic registration of medical images, and is applied to magnetic resonance images of the brain. Experimental results demonstrate very high accuracy in superposition of images from different subjects. There are two major novelties in the proposed algorithm. First, it uses an attribute vector, i.e., a set of geometric moment invariants (GMIs) that are defined on each voxel in an image and are calculated from the tissue maps, to reflect the underlying anatomy at different scales. The attribute vector, if rich enough, can distinguish between different parts of an image, which helps establish anatomical correspondences in the deformation procedure; it also helps reduce local minima, by reducing ambiguity in potential matches. This is a fundamental deviation of our method, referred to as the hierarchical attribute matching mechanism for elastic registration (HAMMER), from other volumetric deformation methods, which are typically based on maximizing image similarity. Second, in order to avoid being trapped by local minima, i.e., suboptimal poor matches, HAMMER uses a successive approximation of the energy function being optimized by lower dimensional smooth energy functions, which are constructed to have significantly fewer local minima. This is achieved by hierarchically selecting the driving features that have distinct attribute vectors, thus, drastically reducing ambiguity in finding correspondence. A number of experiments demonstrate that the proposed algorithm results in accurate superposition of image data from individuals with significant anatomical differences. PMID- 12575880 TI - Flux-based anisotropic diffusion applied to enhancement of 3-D angiogram. AB - We present a new approach to anisotropic diffusion based on a multidirectional diffusion flux. The diffusion flux is decomposed in an orthogonal basis, effectively enabling enhancement of contours as well as diffusion along the contours. To this end, we have selected a three-dimensional basis that depicts the directions of principal curvature and has an interesting interpretation in the context of the vessels. The diffusion function associated to each vector of the basis depends on the first-order derivative of the intensity in this direction, instead of the traditional norm of the smoothed gradient. Accordingly, we present the results of a restoration of computed tomography data of the liver. PMID- 12575881 TI - The role of the medical consultant. AB - The basic concepts of medical consultation have been reviewed. The referring physician and the consultant both have responsibilities to fulfill in order to maximize the effectiveness of the consultation in improving patient care. The reasons for and urgency of the consultation need to be communicated to and understood by the consultant. The consultant needs to respond by promptly evaluating the patient, concisely documenting his findings, and communicating his recommendations to the referring physician. As described by Bates, the ideal medical consultant will "render a report that informs without patronizing, educates without lecturing, directs without ordering, and solves the problem without making the referring physician appear to be stupid". The consultant should try to support the referring physician and comfort the patient. By following these guidelines, the consultant will be more effective in providing useful, informative advice likely to result in enhanced compliance with the recommendations and improved patient outcome. PMID- 12575882 TI - The case against routine preoperative laboratory testing. AB - In this article, we have shown that almost all "routine" laboratory tests before surgery have limited clinical value. Clinicians should order only a small number of routine tests based on age as noted in Table 13. Selective use of other preoperative tests should be based on history and physical examination findings that identify subgroups of patients who are more likely to have abnormal results. In general, clinicians should order tests only if the outcome of an abnormal test will influence management. When an abnormal test results from such testing, it is critical that physicians document their thinking about the result. Most routine preoperative tests are neither expensive nor risky. For this reason, clinicians can have a low threshold for ordering these tests in patients for whom the frequency of abnormalities is increased compared with a healthy population. We believe that physicians should not be criticized for selective test ordering before surgery. Physicians and institutions recommending routine preoperative testing for all patients provide no clinical value to their patients at considerable cost. PMID- 12575883 TI - Perioperative medication management. AB - One of the consultant's roles is to make recommendations regarding the use of medications in the perioperative period. Unfortunately, the data in this area are often insufficient to provide evidence-based recommendations. In this article, we have provided advice considering the pharmacokinetics of the drug, the effect on the primary disease of stopping medications, and the effect of the medication on perioperative risk, including potential drug interactions with anesthetic agents. PMID- 12575884 TI - Antimicrobial prophylaxis in the surgical patient. AB - The primary prophylactic measure against postoperative infection is antiseptic technique in patient preparation, during surgery, and in postoperative patient care. Antimicrobial prophylaxis against postoperative infection is not indicated for procedures with a low infection rate because the expected benefit of antimicrobial treatment is less than the risk of an adverse medication reaction. Antimicrobial prophylaxis has been demonstrated to be of greater benefit than risk in some procedures with higher infection rates; however, because the problem is complex and the data are limited, extra-polating these findings to the practitioner's setting and the individual patient remains a challenge (Table 1). Although antimicrobial prophylaxis for bacterial endocarditis is not effective for most patients, the seriousness of the potential infection has driven the creation of guidelines recommending prophylaxis for at-risk patients undergoing at-risk procedures. Applying these guidelines appropriately could help to reduce unwarranted use of antimicrobials. In the prophylactic use of antimicrobials, as in many medical interventions, the difficulty is balancing the risks of the intervention with the potential benefits. Although we do not have either the randomized, controlled trials or the detailed, patient-specific information to estimate this balance precisely, there are general guidelines to help the clinician choose treatment for most patients. PMID- 12575885 TI - DVT prophylaxis and anticoagulation in the surgical patient. AB - One of the most common postoperative complications is venous thromboembolism, a term encompassing deep vein thrombosis and pulmonary embolism. This article reviews the epidemiology, natural history, difficulties in diagnosis, and strategies for the prevention of postoperative venous thromboembolism. We thoroughly review the currently available methods for thromboprophylaxis including: early ambulation, elastic compression stockings, pneumatic compression devices, inferior vena cava filters, and a variety of pharmacologic agents such as unfractionated heparin, warfarin, aspirin, low molecular weight heparin, and pentasaccharides. Finally, we review the perioperative management of patients on long-term oral anticoagulation. PMID- 12575886 TI - Preoperative risk evaluation and perioperative management of patients with coronary artery disease. AB - We have reviewed the methods of evaluating a patient's cardiac risk preoperatively using a careful history, physical examination, and EKG. Based on this information, various risk indices, guidelines, and algorithms can further assist the physician in deciding which patients can undergo surgery without further testing and which patients might benefit from further cardiac evaluation or medical therapy prior to surgery. The physician must keep in mind that a test should not be ordered if it is unlikely to alter the patient's management, and it is rarely necessary to perform a revascularization procedure with the sole purpose of getting a patient through surgery. Ongoing research is likely to lead to improvement in perioperative medical therapy. PMID- 12575887 TI - Preoperative assessment and perioperative management of the patient with nonischemic heart disease. AB - We have reviewed important issues relating to hypertension, congestive heart failure, arrhythmias and conduction defects, and valvular heart disease in caring for the patient with nonischemic heart disease in the perioperative period. Careful assessment by history and physical examination along with targeted testing will allow the clinician to identify potential complications, provide guided medical therapy, and better utilize other resources to reduce perioperative risk. PMID- 12575888 TI - Preoperative evaluation for postoperative pulmonary complications. AB - Preoperative risk assessment for postoperative pulmonary complications is essential when counseling patients about the risks of surgery because of their significant associated morbidity and mortality. There are many patient-related, operation-related, and anesthesia-related risk factors for the development of PPCs. Though many of these risk factors are not modifiable, they can be useful in evaluating preoperative risk, especially when combined into formal risk indices. Preoperative risk assessment enables clinicians to target preoperative testing and perioperative risk reduction strategies to high-risk patients. Reducing PPC risk at the patient level will require a greater understanding of the impact of modifying risk factors through interventional trials. Reducing hospital PPC rates will require future research into the processes of care associated with PPCs through controlled observational and interventional trials. PMID- 12575889 TI - Perioperative evaluation and management of the patient with endocrine dysfunction. AB - Whenever possible, endocrine disorders should be identified and evaluated prior to surgery. A plan for perioperative management of diabetes should be based on the type of diabetes, what diabetes medications are taken, the status of diabetes control, and what type of surgery is planned. Perioperative management of diabetes must include bedside glucose monitoring. Patients with mild hypothyroidism can safely proceed with elective surgery. Elective surgery should be postponed for patients with moderate or severe hypothyroidism. Patients who have mild hyperthyroidism can undergo elective surgery with preoperative beta blockade. Elective surgery should not be done on patients with moderate or severe hyperthyroidism until they are euthyroid. Patients with pheochromocytoma need to be identified and properly treated before surgery to prevent perioperative cardiovascular complications. Patients who take endogenous steroids should have the status of their HPA axis determined prior to surgery. If the patient is undergoing moderate or major surgical stress and has documented or presumed HPA suppression, then stress doses of steroids should be give perioperatively. PMID- 12575890 TI - Perioperative care of the patient with renal failure. AB - Preventing postoperative ARF, especially in subjects with pre-existing chronic kidney disease, and caring for ESRD patients undergoing surgery are challenging and best accomplished by a team comprised of primary care physician, nephrologist, cardiologist, surgeon, anesthesiologist, endocrinologist, and nutritionist. Elimination of risk factors for ARF whenever possible, as well as early diagnosis, may improve the outcome of this devastating illness. Drugs capable of preventing or changing the course of postoperative ARF may be available soon. For uremic patients, a comprehensive approach is necessary to minimize morbidity and mortality imposed by numerous comorbid conditions. PMID- 12575891 TI - Surgery in the patient with liver disease. AB - Management of the surgical patient with liver disease begins with a careful preoperative assessment (Fig. 1). Any clues to liver disease on history and physical examination should be investigated to ascertain the cause of the clinical finding. More data on surgical patients with unexpected liver disease are now available. Patients undergoing emergent surgery are at significant risk of developing liver dysfunction. Child's class still correlates strongly to postoperative complications. Cornerstones of perioperative management in these patients are medical treatment of complications of chronic liver disease, such as ascites; coagulopathy; prevention of encephalopathy; and rapid treatment of dangerous postoperative complications, such as acute acalculous cholecystitis. Evolving knowledge of the effects of anesthesia, improving surgical techniques, and use of better diagnostic tests will help in the reduction of perioperative complications in these patients. PMID- 12575892 TI - Evaluation and management of anemia and bleeding disorders in surgical patients. AB - The perioperative period offers a unique hemostatic and physiologic challenge. Evaluation of anemia and the decision to transfuse play an important role in the perioperative period. Achievement of adequate hemostasis is important. A bleeding oriented history and physical, along with some baseline tests, may help alert the physician to the possibility of a bleeding disorder. Finally, some patients may need correction of their bleeding disorder before surgery or careful monitoring in the perioperative period. PMID- 12575893 TI - Perioperative issues in patients with cancer. AB - The perioperative care of patients with cancer can be an exciting challenge. The physician must consider many factors, including the cancer diagnosis, the extent of disease, treatment received, the presence of comorbid conditions, and the patient's prognosis and must understand the impact of these factors on the planned surgical procedure. In this setting, the physician has the opportunity to perform an essential role in the perioperative management of patients with cancer. PMID- 12575894 TI - Surgery in the patient with neurologic disease. AB - Patients with neurologic disease who require surgery present distinct issues and challenges for the medical consultant. Although it is not possible to offer a unified approach to neurologic patients, the primary care consultant should understand the clinical issues that are common to these patients, and the individual considerations necessitated by the nature of the neurologic disorder and the clinical characteristics of the patient. The preoperative evaluation combines elements of literature evidence on risk assessment with a thorough understanding of the planned procedure and local practice patterns, and clinical judgment as to the estimated risk-benefit ratio. Perioperative management necessitates attention to many general principles of perioperative care, such as awareness of the potential for cardiopulmonary complications and the need for DVT prophylaxis. In addition, there are management issues for neurologic patients, such as blood pressure control and evaluation of hyponatremia, which may differ from other surgical patients. In these circumstances, the interaction of the neurologic condition with the medical condition and the implications of treatment on the underlying neurologic process also need to be considered. PMID- 12575895 TI - Perioperative care for the elderly patient. AB - As the population survives longer, surgery has become a much more common consideration. Preoperative management of these patients requires a working knowledge of changes associated with aging and the physiology of surgery and anesthesia. Using this information, patients can be clinically evaluated effectively and plans made for their perioperative care to minimize complications. PMID- 12575896 TI - Biosynthesis of 2-acetamido-2,6-dideoxy-L-hexoses in bacteria follows a pattern distinct from those of the pathways of 6-deoxy-L-hexoses. AB - 6-Deoxy-L-hexoses have been shown to be synthesized from dTDP-D-glucose or GDP-D mannose so that the gluco/galacto-configuration is converted into the manno/talo configuration, and manno/talo is switched to gluco/galacto. Our laboratory has been investigating the biosynthesis of 2-acetamido-2,6-dideoxy-L-hexoses in both Gram-positive and Gram-negative bacteria, and in a recent paper we described the biosynthesis of the talo (pneumosamine) and galacto (fucosamine) derivatives from UDP-D-N-acetylglucosamine a 2-acetamido sugar [Kneidinger, O'Riordan, Li, Brisson, Lee and Lam (2003) J. Biol. Chem. 278, 3615-3627]. In the present study, we undertake the task to test the hypothesis that UDP-D-N-acetylglucosamine is the common precursor for the production of 2-acetamido-2,6-dideoxy-L-hexoses in the gluco-, galacto-, manno- and talo-configurations. We present data to reveal the steps for the biosynthesis of the gluco (quinovosamine)- and manno (rhamnosamine)-configured compounds. The corresponding enzymes WbvB, WbvR and WbvD from Vibrio cholerae serotype O37 have been overexpressed and purified to near homogeneity. The enzymic reactions have been analysed by capillary electrophoresis and NMR spectroscopy. Our data have revealed a general feature of reaction cascades due to the three enzymes. First, UDP-D-N-acetylglucosamine is catalysed by the multi-functional enzyme WbvB, whereby dehydration occurs at C-4, C-6 and epimerization at C-5, C-3 to produce UDP-2-acetamido-2,6-dideoxy-L-lyxo-4 hexulose. Secondly, this intermediate is converted by the C-4 reductase, WbvR, in a stereospecific reaction to yield UDP-2-acetamido-L-rhamnose. Thirdly, UDP-2 acetamido-L-rhamnose is epimerized at C-2 to UDP-2-acetamido-L-quinovose by WbvD. Interestingly, WbvD is also an orthologue of WbjD, but not vice versa. Incubation of purified WbvD with UDP-2-acetamido-2,6-dideoxy-L-talose and analysing the reaction products by capillary electrophoresis revealed the same product peak as when WbjD was used. This sugar nucleotide is a specific substrate for WbjD and is a C-4 epimer of UDP-2-acetamido-L-rhamnose. PMID- 12575897 TI - Low NO bioavailability in CCl4 cirrhotic rat livers might result from low NO synthesis combined with decreased superoxide dismutase activity allowing superoxide-mediated NO breakdown: A comparison of two portal hypertensive rat models with healthy controls. AB - BACKGROUND: In cirrhotic livers, the balance of vasoactive substances is in favour of vasoconstrictors with relatively insufficient nitric oxide. Endothelial dysfunction has been documented in cirrhotic rat livers leading to a lower activity of endothelial nitric oxide synthase but this might not be sufficient to explain the low nitric oxide presence. We compared the amount of all nitric oxide synthase isoforms and other factors that influence nitric oxide bioavailability in livers of two portal hypertensive rat models: prehepatic portal hypertension and carbon tetrachloride induced cirrhosis, in comparison with healthy controls. RESULTS: Endothelial nitric oxide synthase was the solely detected isoform by Western blotting in all livers. In cirrhotic livers, the amount of endothelial nitric oxide synthase protein was lower than in healthy controls, although an overlap existed. Levels of caveolin-1 messenger RNA were within the normal range but endothelin-1 messenger RNA levels were significantly higher in cirrhotic livers (p < 0.05). A markedly lower superoxide dismutase activity was observed in cirrhotic livers as compared to healthy controls (p < 0.05). CONCLUSIONS: In contrast to prehepatic portal hypertension, cirrhotic livers had decreased endothelial nitric oxide synthase protein and enhanced endothelin-1 messenger RNA amount. We hypothesise that a vasodilator/vasoconstrictor imbalance may be further aggravated by the reduced activity of superoxide dismutase. Decreased activity allows enhanced superoxide action, which may lead to breakdown of nitric oxide in liver sinusoids. PMID- 12575900 TI - Fate of genetically modified maize DNA in the oral cavity and rumen of sheep. AB - The polymerase chain reaction (PCR) technique was used to investigate the fate of a transgene in the rumen of sheep fed silage and maize grains from an insect resistant maize line. A 1914-bp DNA fragment containing the entire coding region of the synthetic cryIA(b) gene was still amplifiable from rumen fluid sampled 5 h after feeding maize grains. The same target sequence, however, could not be amplified from rumen fluid sampled from sheep fed silage prepared from the genetically modified maize line. PCR amplification of a shorter (211-bp), yet still highly specific, target sequence was possible with rumen fluid sampled up to 3 and 24 h after feeding silage and maize grains, respectively. These findings indicate that intact transgenes from silage are unlikely to survive significantly in the rumen since a DNA sequence 211-bp long is very unlikely to transmit genetic information. By contrast, DNA in maize grains persists for a significant time and may, therefore, provide a source of transforming DNA in the rumen. In addition, we have examined the biological activity of plasmid DNA that had previously been exposed to the ovine oral cavity. Plasmid extracted from saliva sampled after incubation for 8 min was still capable of transforming competent Escherichia coli to kanamycin resistance, implying that DNA released from the diet within the mouth may retain sufficient biological activity for the transformation of competent oral bacteria. PMID- 12575901 TI - Effect of feed intake on amino acid transfers across the ovine hindquarters. AB - Responses in variables of amino acid (AA) metabolism across peripheral tissues to feed intake were studied in six sheep (mean live weight 32 kg) prepared with arterio-venous catheters across the hindquarters. Four intakes (0.5, 1.0, 1.5 and 2.5 x maintenance energy) were offered over 2-week periods to each sheep in a Latin square design with two animals replicated. Animals were infused intravenously with a mixture of U-13C-labelled AA for 10 h and integrated blood samples withdrawn from the aorta and vena cava hourly between 5 and 9 h of infusion. Biopsy samples were also taken from skin and m. vastus lateralis. Data from both essential (histidine, isoleucine, leucine, lysine, phenylalanine, threonine) and nonessential (glycine, proline, serine, tyrosine) AA were modelled to give rates of inward and outward transport, protein synthesis and degradation, plus the fraction of total vascular inflow that exchanged with the hindquarter tissues. Rates of inward transport varied more than 10-fold between AA. For all essential AA (plus serine), inward transport increased with food intake (P<0.04). There were corresponding increases in AA efflux (P<0.05) from the tissues for threonine and the branched-chain AA. Protein synthesis rates estimated from the kinetics of these AA also increased with intake (P<0.02). Rates of inward transport greatly exceeded the amount of AA necessary to support protein retention, but were more similar to rates of protein synthesis. Nutritional or other strategies to enhance AA transport into peripheral tissues are unlikely to increase anabolic responses. PMID- 12575902 TI - Effect of the addition of malate on in vitro rumen fermentation of cereal grains. AB - Batch cultures of mixed rumen micro-organisms were used to study the effects of different concentrations of malate (Rumalato(R); Norel & Nature S.A., Barcelona, Spain; composed of disodium malate-calcium malate (0.16:0.84, w/w)) on the fermentation of four cereal grains (maize, barley, wheat and sorghum). Rumen contents were collected from four Merino sheep fed lucerne hay ad libitum and supplemented with 300 g concentrate/d. Rumalato(R) was added to the incubation bottles to achieve final concentrations of 0, 4, 7 and 10 mm-malate. Gas production was measured at regular intervals up to 120 h. Malate increased (P<0.01) the average fermentation rate of all substrates, and the lag time decreased (P<0.05) linearly with increasing concentrations of malate for all substrates, with the exception of sorghum. In 17 h incubations, the final pH and total volatile fatty acid production increased (P<0.001) linearly for all substrates as malate concentration increased from 0 to 10 mm. Propionate and butyrate production increased (P<0.05), while the value of the acetate : propionate ratio and l-lactate concentrations decreased (P<0.05) linearly with increasing doses of malate. Malate treatment increased (P<0.05) the CO2 production and decreased the production of CH4, although this effect was not significant (P>0.05) for maize. Malate at 4 and 7 mm increased (P<0.05) optical density of the cultures measured at 600 nm for maize, with no differences for the other substrates. The results indicate that malate may be used as a feed additive for ruminant animals fed high proportions of cereal grains, because it increased pH and propionate production and decreased CH4 production and l-lactate concentrations; however, in general, no beneficial effects of 10 compared with 7 mm-malate were observed. PMID- 12575899 TI - Lipoxygenase and cyclooxygenase metabolism: new insights in treatment and chemoprevention of pancreatic cancer. AB - The essential fatty acids, linoleic acid and arachidonic acid play an important role in pancreatic cancer development and progression. These fatty acids are metabolized to eicosanoids by cyclooxygenases and lipoxygenases. Abnormal expression and activities of both cyclooxygenases and lipoxygenases have been reported in pancreatic cancer. In this article, we aim to provide a brief summary of (1) our understanding of the roles of these enzymes in pancreatic cancer tumorigenesis and progression; and (2) the potential of using cyclooxygenase and lipoxygenase inhibitors for pancreatic cancer treatment and prevention. PMID- 12575903 TI - The combined effects of garlic oil and fish oil on the hepatic antioxidant and drug-metabolizing enzymes of rats. AB - This present study was designed to investigate the combined modulatory effect of garlic oil (GO) and fish oil (FO) on the antioxidant and drug metabolism systems. Rats were fed either a low-maize oil (MO) diet (50 g MO/kg), high-MO diet (235 g MO/kg) or high-FO diet (205 g FO+ 30 g MO/kg) and received different doses of GO (0-200 mg/kg body weight) three times per week for 6 weeks. Fatty acid analysis showed that 20 : 5n-3 and 22 : 6n-3 were incorporated into serum lipid at the expense of 18 : 2n-6 and 20 : 4n-6 in rats fed the high-FO diet. GO dose dependently increased hepatic glutathione S-transferase (GST), glutathione reductase, superoxide dismutase (SOD) and ethoxyresorufin O-deethylase (EROD) activities, but decreased glutathione peroxidase and N-nitrosodimethylamine demethylase (NDMAD) activities (P<0.05). With the exception of glutathione peroxidase, the activities of glutathione reductase, SOD, GST, EROD and NDMAD were modulated by the dietary fat. The high-FO group had greater SOD and EROD activity than either MO-fed group; it also had greater NDMAD activity than the low-MO group (P<0.05). GST activity was higher in rats fed high-FO or high-MO diets than rats fed the low-MO diet. Change in erythromycin demethylase activity, however, was not caused by either dietary fat or GO. Immunoblot assay showed that GO dose-dependently enhanced the protein level of the Ya, Yb1, Yc isoenzymes of GST and cytochrome P450 (CYP) 1A1 and 3A1, but GO suppressed CYP2E1 expression. Regardless of the dosage of GO, the high-FO diet increased CYP1A1, CYP3A1 and CYP2E1 levels compared with the high- and low-MO diets. Accompanying the changes observed in immunoblots, CYP1A1 and CYP3A1 mRNA levels were increased by GO in a dose-dependent manner and also increased additively in combination with FO feeding. These present results indicate that co-administration of GO and FO modulates the antioxidant and drug-metabolizing capacity of animals and that the effect of GO and FO on drug-metabolizing enzymes is additive. PMID- 12575904 TI - Dietary rhubarb (Rheum rhaponticum) stalk fibre does not lower plasma cholesterol levels in diabetic rats. AB - Rhubarb (Rheum rhapontiam) stalk fibre was previously shown to be hypolipidaemic under clinical and experimental conditions. The present study was undertaken to investigate whether rhubarb stalk fibre has a hypolipidaemic effect under diabetic conditions. Two models of diabetic rats were used: streptozotocin induced diabetic rats, and diabetes-prone BB (BBdp) rats. The plasma cholesterol and triacylglycerol concentrations were elevated after the onset of diabetes in BBdp rats, but not in sterptozotocin-induced diabetic rats. The rhubarb-fibre diet had no effect on the plasma cholesterol or triacylglycerol concentrations of diabetic rats. The hypolipidaemic effect of rhubarb stalk fibre has been suggested to be due to the bile-acid-binding capacity of rhubarb fibre, which in turn up regulates cholesterol 7alpha-hydroxylase (cyp7a) activity. cyp7a is the first and the rate-limiting enzyme in the breakdown of cholesterol to bile acids. We measured the cyp7a activity and mRNA levels in control and diabetic rats fed rhubarb- and cellulose-fibre diets. The cyp7a activity and mRNA abundance were increased in both diabetic rat models, indicating that bile acid synthesis is enhanced in diabetes. Feeding a diet enriched with rhubarb fibre caused a slight but significant increase (P<0.05) in cyp7a enzyme activity in BBdp rats, but no change in cyp7a mRNA abundance was detected. These results suggest that although a rhubarb-fibre-enriched diet increased cyp7a activity in BBdp rats, there was no apparent therapeutic benefit in terms of lowering plasma cholesterol concentrations. PMID- 12575905 TI - A randomised four-intervention crossover study investigating the effect of carbohydrates on daytime profiles of insulin, glucose, non-esterified fatty acids and triacylglycerols in middle-aged men. AB - Postprandial concentrations of glucose, insulin and triacylglycerols (TG) correlate to risk for CHD. Carbohydrates affect many metabolites that could have a potential effect on cardiovascular risk factors. The objective of the present study was to examine, using a randomised prospective study, the acute (day 1) and ad libitum medium-term (day 24) effects of four diets: a high-fat diet (HIGH-FAT; 50 % fat, >34 % monounsaturated fatty acids); a low-glycaemic index (GI) diet (LOW-GI; high-carbohydrate, low-GI); a high-sucrose diet (SUCROSE; high carbohydrate increase of 90 g sucrose/d); a high-GI diet (HIGH-GI; high carbohydrate, high-GI). Daytime profiles (8 h) (breakfast, lunch and tea) of lipid and carbohydrate metabolism were completed during day 1 and day 24. Seventeen middle-aged men with one or more cardiac risk factors completed the study. There was no change from day 1 or between diets in fasting glucose, lipids or homeostatic assessment model (HOMA) on day 24. The HIGH-FAT compared with the three high-carbohydrate diets was associated with lower postprandial insulin and glucose but higher postprandial TG and non-esterified fatty acids (NEFA). There was a significant increase in the 6 h (15.00 hours) TG concentration (day 1, 2.6 (sem 0.3) mmol/l v. day 24, 3.3 (sem 0.3) mmol/l; P<0.01) on the SUCROSE diet. Postprandial HOMA (i.e. incremental area under the curve (IAUC) glucose (mmol/l per min)xIAUC insulin/22.5 (mU/l per min)) median changes from day 1 to day 24 were -61, -43, -20 and +31 % for the HIGH-FAT, LOW-GI, SUCROSE and HIGH-GI diets respectively. The HIGH-GI percentage change was significantly different from the other three diets (P<0.001). Despite being advised to maintain an identical energy intake there was a significant weight change (-0.27 (sem 0.3) kg; P<0.02) on the LOW-GI diet compared with the SUCROSE diet (+0.84 (sem 0.3) kg). In conclusion the HIGH-FAT diet had a beneficial effect on postprandial glucose and insulin over time but it was associated with higher postprandial concentrations of TG and NEFA. Conversely the HIGH-GI diet appeared to increase postprandial insulin resistance over the study period. PMID- 12575906 TI - Conjugated linoleic acid decreases fat accretion in pigs: evaluation by dual energy X-ray absorptiometry. AB - Thirty female Large White x Landrace pigs (average weight 57.2 (sd 1.9) kg) were allocated to one of six dietary treatments containing 0, 1.25, 2.5, 5.0, 7.5 or 10.0 g 55 % conjugated linoleic acids (CLA) isomers (CLA-55)/kg diet and fed for 8 weeks. Each pig was scanned at 0, 28 and 56 d and again at post slaughter using dual-energy X-ray absorptiometry (DXA) to determine the temporal pattern of body composition responses. Values determined by DXA were adjusted using regression equations generated from validation experiments between chemically and DXA predicted values. Overall, there was a significant linear reduction in fat content with the increasing levels of CLA in the diet (P=0.007, P=0.011, P=0.008 at week 4, week 8 and for the carcass, respectively). The greatest improvement was recorded at the early stages of CLA supplementation and for the highest dose of CLA (week 4, -19.2 % compared with week 8, -13.7 %). In the first 4 weeks of feeding CLA, pigs receiving 10 g CLA-55/kg diet deposited 93 g less fat/d than pigs fed basal diets (P=0.002) compared with only 6 g less fat than control animals in the final 4 weeks. Lean content and lean deposition rate were maximised at 5 and 2.5 g CLA-55/kg diet for the first 4 weeks (P=0.016) and the final 4 weeks of treatment respectively. DXA estimates of bone mineral content and bone mineral density were not affected by CLA supplementation throughout the experiment. These data demonstrate that dietary CLA decreases body fat in a dose dependent manner and that the response is greatest over the initial 4 weeks of treatment. PMID- 12575907 TI - The hepatic retinyl ester hydrolase activity is depressed at the onset of diabetes in BB rats. AB - Dietary vitamin A as retinyl ester is hydrolysed and re-esterified with long chain fatty acids in the small intestine. The esterified vitamin A is subsequently stored in the liver, where it is hydrolysed to free retinol to be transported by carrier proteins to the target tissue. A decreased availability of retinol carrier proteins has been suggested to be responsible for affecting metabolic availability of vitamin A in type 1 diabetes. Using BB Wistar rats, the present study was undertaken to examine whether the presence of a hyperglycaemic state modifies retinyl ester hydrolase (REH) activity in the intestine and the liver. At the onset of diabetes, hepatic REH enzymatic activity was significantly decreased. However, REH activity remained unaffected in the small intestine, including both ileum and jejunum. Diabetes also resulted in decreased plasma and liver concentrations of retinol. An in vitro study was conducted to examine the effect of diabetes on the intestinal uptake of retinyl palmitate. Jejunum and ileum from diabetic and non-diabetic BB rats were incubated with labelled retinyl palmitate at different concentrations ranging from 32 to 256 nmol/l. The uptake of retinyl palmitate was increased in both diabetic and non-diabetic rats together with the increase of substrate concentration. However, no significant difference was observed in the uptake of retinyl palmitate between diabetic and non-diabetic rats. These present results suggest that the depressed hepatic REH activities may contribute to the diabetes-associated metabolic derangement of vitamin A. PMID- 12575908 TI - Casein and whey exert different effects on plasma amino acid profiles, gastrointestinal hormone secretion and appetite. AB - Protein, generally agreed to be the most satiating macronutrient, may differ in its effects on appetite depending on the protein source and variation in digestion and absorption. We investigated the effects of two milk protein types, casein and whey, on food intake and subjective ratings of hunger and fullness, and on postprandial metabolite and gastrointestinal hormone responses. Two studies were undertaken. The first study showed that energy intake from a buffet meal ad libitum was significantly less 90 min after a 1700 kJ liquid preload containing 48 g whey, compared with an equivalent casein preload (P<0.05). In the second study, the same whey preload led to a 28 % increase in postprandial plasma amino acid concentrations over 3 h compared with casein (incremental area under the curve (iAUC), P<0.05). Plasma cholecystokinin (CCK) was increased by 60 % (iAUC, P<0.005), glucagon-like peptide (GLP)-1 by 65 % (iAUC, P<0.05) and glucose dependent insulinotropic polypeptide by 36 % (iAUC, P<0.01) following the whey preload compared with the casein. Gastric emptying was influenced by protein type as evidenced by differing plasma paracetamol profiles with the two preloads. Greater subjective satiety followed the whey test meal (P<0.05). These results implicate post-absorptive increases in plasma amino acids together with both CCK and GLP-1 as potential mediators of the increased satiety response to whey and emphasise the importance of considering the impact of protein type on the appetite response to a mixed meal. PMID- 12575909 TI - Effects of ornithine alpha-ketoglutarate on insulin secretion in rat pancreatic islets: implication of nitric oxide synthase and glutamine synthetase pathways. AB - Ornithine alpha-ketoglutarate (OKG) administration in human subjects elicits insulin secretion. We investigated whether this action was related to an effect of OKG on islets of Langerhans, and addressed the underlying mechanisms of action. For this purpose the influence of OKG on insulin secretion was measured in isolated rat islets of Langerhans under two different conditions. In incubated islets, OKG (0.25 to 2.5 mmol/l) significantly and dose-relatedly increased insulin secretion (1.7- to 4.2-fold; P<0.05 v. basal). To study the kinetics of OKG-stimulated insulin secretion, perifusion experiments were performed, which showed that OKG affected insulin secretion in both initial and later phases. Experiments using alpha-ketoglutarate (alpha-KG) (1 mmol/l) or ornithine (Orn) (2 mmol/l) alone, in concentrations equal to that of OKG, showed that the OKG induced insulin secretion could not be obtained by either component alone, suggesting that an alpha-KG-Orn interaction is mandatory for the insulin secreting effect to occur. Since data obtained in vivo suggest that effects of OKG may depend on the synthesis of NO, glutamine and/or polyamines, three metabolic pathways potentially involved in insulin secretion, we then evaluated their contribution by means of their respective inhibitors: l-NG-nitroarginine methyl ester (l-NAME), methionine sulfoximine (MSO) and difluoromethylornithine (DFMO). Both l-NAME and MSO were able significantly to reduce OKG-induced insulin secretion (30 and 40 % respectively; P<0.05), while DFMO was ineffective. Thus OKG is an effective stimulator of insulin secretion, requiring the joint presence of both Orn and alpha-KG, and acting mainly via the synthesis of NO and glutamine. A better understanding of OKG insulino-secretory properties and its mechanisms of action are a prerequisite for its use in insulin-compromised situations. PMID- 12575910 TI - The effect of various vitamin E derivatives on the urokinase-plasminogen activator system of ovine macrophages and neutrophils. AB - The effect of vitamin E derivatives on the urokinase-plasminogen activator (u-PA) system of resting and phorbol myristate acetate (PMA)-activated ovine macrophages and neutrophils were investigated. Blood monocyte-macrophages and neutrophils were isolated from twenty-four animals. Macrophages or neutrophils were cultured in vitro for 3 or 24 h with or without various vitamin E derivatives: free alpha tocopherol (alpha-T), alpha-tocopheryl acetate (alpha-TA), or alpha-tocopheryl succinate (alpha-TS). Following incubation, cells were stimulated with 80 microm PMA. Total cell-associated u-PA, membrane-bound u-PA and free u-PA binding sites were determined before and after stimulation with PMA. Results showed that none of the vitamin E derivatives had any effect (P>0.05) on the u-PA system of resting monocyte-macrophages or neutrophils. In contrast, alpha-TS, but not alpha TA or alpha-T, increased (P<0.01) total cell-associated u-PA and membrane-bound u PA of PMA-stimulated macrophages and neutrophils. alpha-TS had no effect (P>0.05) on total u-PA and membrane-bound u-PA activities of macrophages and neutrophils cultured in the presence of 4-phorbol 12,13 didecanoate, a phorbol ester that does not activate protein kinase (PK) C. Addition of H7 (1-(5 isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride), which is a potent inhibitor of both PK A and C, completely abolished the effect of alpha-TS on total cell-associated u-PA and membrane-bound u-PA of PMA-activated macrophages and neutrophils. Addition of HA1004 (N-(2-quanidinoethyl)-5-isoquinoline sulfonamide hydrochloride), which is a potent PK A but a weak PK C inhibitor, had no effect (P>0.05) on total cell-associated u-PA and membrane-bound u-PA of PMA activated macrophages and neutrophils cultured in the presence of alpha-TS. Thus, PK C modulates the effect of alpha-TS on the u-PA system of ovine macrophages and neutrophils. PMID- 12575911 TI - Serum fatty acid levels, dietary style and coronary heart disease in three neighbouring areas in Japan: the Kumihama study. AB - CHD mortality is extremely low in Japan, particularly in rural districts, when compared with that in Western countries. This has been partly attributed to the difference in dietary lifestyle. We investigated the factors influencing CHD mortality in a rural coastal district of Japan, comprising mercantile, farming, and fishing areas with distinct dietary habits. We prospectively examined the incidence of CHD from 1994 to 1998, as well as coronary risk factors and serum fatty acid concentrations. The incidence of angina pectoris was significantly (P=0.01) lower in the fishing area than in the mercantile and farming areas. Blood pressure, physical activity, prevalence of diabetes, serum levels of uric acid and HDL-cholesterol were similar between the three areas. Total- and LDL cholesterol levels were significantly lower but the smoking rate was markedly higher in the fishing area than in the other two areas. Serum levels of saturated fatty acids and n-6 polyunsaturated fatty acids (PUFA) were lowest in the fishing area, but n-3 PUFA did not differ significantly. The n-6:n-3 PUFA ratio was lowest and eicosapentaenoic:arachidonic acid was highest in the fishing area. Although many previous studies have emphasized the beneficial effect of n-3 PUFA in preventing CHD, the present study indicated that a lower intake of n-6 PUFA and saturated fatty acids has an additional preventive effect on CHD even when the serum level of n-3 PUFA is high because of high dietary fish consumption. PMID- 12575912 TI - Demographic and lifestyle characteristics of functional food consumers and dietary supplement users. AB - Functional foods and/or supplements may be used in the context of a healthy lifestyle or as a means to compensate for an unhealthy lifestyle. Adverse long term and/or cumulative effects of functional food or supplement intake are of public health concern; it is therefore important to identify functional food and supplement users. The present study compared Dutch functional food and supplement consumers with non-consumers with regard to demographic and lifestyle factors. The consumption of the most common functional foods and supplements in 2000 was studied (yoghurt with extra lactic acid bacteria, cholesterol-lowering margarine, lemonade and sweets with extra vitamins and minerals, milk and margarine with extra Ca, Ca tablets, multivitamin and mineral supplements, and Echinacea supplements). Data were obtained from self-administered questionnaires filled in by a consumer panel, aged 19-91 years (response rate 76 %, n 1183), representative of the Dutch population. The number of daily consumers of functional foods or supplements appeared to be relatively low (daily use of multivitamin and mineral supplements, 20 %; all other products, 3-9 %). Explanatory variables depended on the type of product; but gender, age, education, and vegetable intake were significant factors in the logistic regression model. Consumption of cholesterol-lowering margarines was more likely to be reported by individuals with a poorer subjective health (odds ratio 2.62 (95 % CI 1.15, 6.05)) and by smokers (odds ratio 2.93 (95 % CI 1.34, 6.40)). In conclusion, determinants of functional food or supplement use depended on the type of product, so generalisation of consumer characteristics over different foods is not legitimate. In addition to research on lifestyle factors, surveys about consumers' attitudes, norms and knowledge regarding functional foods in relation to actual dietary patterns and health risk profiles are necessary. PMID- 12575913 TI - Diets enriched with cereal brans or inulin modulate protein kinase C activity and isozyme expression in rat colonic mucosa. PMID- 12575915 TI - Oxidative stress and apoptosis in immune diseases. AB - Antigenic stimuli increase ROS that influence T-cell activation by interfering with the oxidant-antioxidant balance. Oxidative stress takes place when excess of ROS production is not counterbalanced by antioxidant mechanisms and bcl-2 gene product that inhibits apoptosis by interacting with mitochondrial superoxide dismutase. ROS Excess induces apoptosis both by activation of NF-kB-dependent genes and DNA damage. The latter has been shown to elicit the activation of poly ADP-ribose transferase and the accumulation of p53, thus determining apoptosis. Additionally, oxidative stress may induce formation of cell membrane oxidized lipids, potent inducers of apoptosis. Oxidative stress is also involved in immune diseases. In AIDS, ROS excess and deficiency of antioxidants lead to apoptosis and virus activation. ROS produced at sites of chronic inflammation, have genotoxic effects. As a consequence, abnormalities of the p53 genes might explain the conversion from an inflammatory phase into autonomous progression of rheumatoid arthritis or other chronic inflammatory disorders. PMID- 12575916 TI - Some aspects of Parasitology and Immunology in general Medicine. AB - The objective of these studies is to review the role of some parasites and their components in inflammation, allergy and immune system. We also report recent results published by others group as well as our own. PMID- 12575917 TI - The role of antileukotrienes in the treatment of asthma. AB - Cysteinyl leukotrienes (Cys-LTs) are mediators released in asthma and are both direct bronchoconstrictors and proinflammatory substances that mediated several steps in the pathophysiology of chronic asthma, including inflammatory cells recruitment, vascular leakage, and possibly airway remodelling. Available evidence from clinical trials and real world experience derived from managing patients with asthma justifies a broader role for antiLTRAs in asthma management than that recommended in the National Asthma Education and Prevention Programm (NAEPP) and National Health Lung and Blood Institute (NHLBI) treatment guidelines. Leukotriene-receptor antagonist drugs (LTRAs) seem to be effective alternatives to inhaled corticosteroids (ICS) either as monotherapy or as adjunctive therapy that reduces the need for higher doses of ICS in patients with mild-to-moderate persistent asthma. LTRAs may be used as adjunctive therapy for al levels of disease severity because they are effective in combination with ICS during long-term maintenance therapy. The agents seem especially effective in preventing aspirin-induced asthma, exercise-induced asthma (EIA) and they may provide an additional advantage of reducing nasal congestion in patients with both asthma and rhinitis. PMID- 12575918 TI - Effect of monocyte chemoattractant protein-1 on murine bone marrow cells: proliferation, colony-forming ability and signal transduction pathway involved. AB - Monocyte chemoattractant protein-1 (MCP-1) plays a crucial role in the migration and activation of leukocytes in both physiological and pathological contexts. In this paper, we report the in vitro effect of MCP-1 on myeloid haematopoiesis. MCP 1-treated murine nonadherent bone marrow cells (NABMCs) were assayed for in vitro proliferation and colony forming ability. It is observed that MCP-1 treatment in vitro caused an enhancement in the proliferation and colony forming ability of the murine NABMCs as compared to the untreated cells. This response was concentration-dependent and most effective at a dose of 100ng/ml MCP-1. In the presence of MCSF (200U/ml), GCSF (200U/ml), GMCSF (200U/ml) or IL-3 (200U/ml), the MCP-1-induced colony forming ability of the NABMCs was significantly augmented, indicating a synergistic effect of MCP-1 with these CSFs. However, irrespective of the CSFs used, MCP-1 stimulated the lineage-restricted differentiation of the murine BMCs into predominantly the granulocytic lineage. NABMCs cultured in medium alone formed minimal colonies. The probable signal transduction mechanism responsible for the MCP-1-induced NABMC proliferation/differentiation was also investigated. The results of the colony forming assay indicate that the protein kinase inhibitors, genistein (10&mgr;g/ml), chelenthryin chloride (10&mgr;M), wortmannin (200nM) and PD98059 (10&mgr;M) significantly blocked the in vitro colony forming ability of the MCP-1 treated NABMCs, while the phosphatase inhibitors, okadaic acid (10nM) and sodium orthovanadate (10&mgr;M) caused an increase in the BMC colony forming ability in response to MCP-1. These data suggests the involvement of the respective protein kinases and phosphatases in the above process. Correlating with this, the role of several signaling molecules likes Lyn, p42/44MAPK, PI3K and STAT5 has also been implicated in the signal cascade of murine NABMC proliferation/differentiation following MCP-1 treatment. PMID- 12575919 TI - N(2)O(2)-induced apoptosis of thymocytes involves mobilization of divalent cations. AB - More than 90% of thymocytes undergo apoptosis while undergoing differentiation in the thymus. Although several factors act in concert to induce thymocyte apoptosis, it remains speculative if reactive oxygen intermediates produced by thymic macrophages may play a role in this process. The present investigation was carried out to determine if H(2)O(2) is capable of inducing apoptosis of thymocytes in vitro. It was observed that H(2)O(2) could induce apoptosis of thymocytes in vitro in a dose and time dependent manner. It was further found that H(2)O(2)-induced thymocyte apoptosis was dependent on the mobilization of divalent cations. The result of this study will help further in the understanding of the mechanism of H(2)O(2)-induced apoptosis PMID- 12575920 TI - Involvement of caspace-3 in the cleavage of terminal transferase. AB - To investigate the in vivo role of caspase-3 in Terminal Transferase metabolism DMSO-treated RPMI-8402, a human pre-T cell line was used. In DMSO treated samples (3)H-dGTP incorporation and TdT phosphorylation occurs after 4 hours of treatment. After 8 hours cells undergo TdT proteolysis in addition to its inactivation. The cleavage of TdT into 32- and 58-KDa proteolytic fragments occurred simultaneously with the activation of Caspase-3, but preceded changes associated with the apoptotic process described after 48 hours of treatment. The Caspase-3 peptide inhibitor V, used as a specific inhibitor, prevented TdT proteolysis prolonging its activity and rescued cells from apoptosis. Our experiments suggest that TdT is a nuclear substrate for Caspase-3, the main apoptotic effector protease in many cell types, and that the cleavage of TdT represents a primary step in a signal cascade leading to pre-T cell apoptosis. PMID- 12575921 TI - Effect of yogurt on the inhibition of an intestinal carcinoma by increasing cellular apoptosis. AB - Yogurt consumption has been associated with decreased risk of colon cancer. The apoptosis induced by yogurt as a mechanism involved in the inhibition of an experimental intestinal tumor induced by 1,2 dimethylhydrazine (DMH) in BALB/c mice, has been studied. An imbalance between apoptosis and mitosis or cell proliferation is believed to underlie colon cancer development and progression. We determined the effect of yogurt feeding in the induction of apoptosis and the relationship between mitosis and apoptosis in mice injected with the carcinogen DMH and in mice injected with DMH and fed with a diet supplemented with yogurt. The percentage of mitotic and apoptotic cells was analyzed every 2 weeks from week 8 to 16. They were analyzed on histological slices from the large intestine by colchicine, or Tunel test for mitosis and apoptosis determination, respectively. An increase in the mitosis during the first 4 weeks of tumor development with apoptosis negative and severe dysphasia, was observed in animals treated with the carcinogen. In those animals that received DMH and were given yogurt, a moderate cell proliferation with a significant increase in the number of apoptotic cells was determined. The increase in the apoptotic activity was also observed in the group of animals that received only a diet supplemented with yogurt. The results suggest that yogurt would influence the balance between mitosis and apoptosis, which was modified during the carcinogenesis process, by increasing cellular apoptosis. Dietary supplementation of yogurt may play a role in modulating cell proliferation or apoptosis during the development of colorectal carcinoma. PMID- 12575922 TI - Fexofenadine in chronic idiopathic urticaria: a clinical and immunohistochemical evaluation. AB - Fexofenadine is a non-sedating selective third-generation antihistamine, which also exerts an anti-inflammatory action. The aim of this study was to evaluate the influence on the expression of inflammatory skin mediators, together with the efficacy and tolerability, of fexofenadine in chronic idiopathic urticaria (CIU). Fexofenadine 180mg was administered once daily for 4 weeks after a placebo run-in phase of 3 to 7 days. Efficacy paramaters were obtained from patients' assessment of urticaria symptoms. Non-lesional skin of patients with active CIU was studied immunohistochemically before and after treatment. The expression of the following mediators was evaluated: adhesion molecules (ICAM-1, ELAM-1, VCAM-1); mast cell proteases (chymase and tryptase) and proinflammatory cytokines (IL-1beta, IL-3, IL-6 and TNF-alpha). Of the 20 subjects enrolled, 3 dropped out of the study. Treatment proved successful in most cases (88.2%) (p <0.01) and a significant improvement of all symptoms was registered. Treatment was well-tolerated by all patients; adverse events, neither serious nor drug-related, occurred in any case. Immunochemistry revealed at the baseline a significant expression of ELAM-1, VCAM 1, tryptase, chymase, and TNF-alpha (p= 0.05) in non-lesional skin of patients compared to normal controls. After treatment with fexofenadine, there was a significant decrease in the expression of ELAM-1 (p= 0.02), VCAM-1 (p= 0.04) and tryptase (p= 0.04), whereas no relevant change was observed for the other parameters examined. This work confirms the efficacy and tolerability of fexofenadine HCl 180mg in CIU. These preliminary data show a trend towards a decrease in the expression of tryptase and some adhesion molecules after treatment, suggesting an anti-inflammatory activity of fexofenadine. PMID- 12575923 TI - Verapamil reduces coronary endothelium damage and cardiomyocyte necrosis but not apoptosis after ischemia and reperfusion: ex-vivo study in rat hearts. AB - We tested the hypothesis of beneficial effects of the calcium-blocker verapamil in a model of ischemia-reperfusion, and investigated its effects against coronary microcirculation and cardiomyocyte apoptosis. Isolated working rat hearts were subjected to 15 min global ischemia and 22-180 min reperfusion in the presence or absence of verapamil (0.25 &mgr;M). We evaluated creatinephosphokinase (CK) in coronary effluent, heart weight changes, microvascular permeability (extravasation of fluoresceine-labeled albumin), ultrastructural alterations, and cardiomyocyte apoptosis (by 1.5% agarose gel electrophoresis and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling technique). In this model, 0.25 &mgr;M verapamil significantly reduced myocardial damage, CK release and vascular hyperpermeability, concomitant with a reduction in endothelial and cardiomyocyte lesions; on the contrary, 0.25 &mgr;M verapamil was unable to reduce cardiomyocyte apoptosis. In conclusion, in the absence of perfusing granulocytes, the acute administration of a pharmacologically relevant verapamil concentration reduces ischemia-reperfusion injury and prevents coronary endothelial cell and cardiomyocyte necrotic cell death but it is unable to reduce apoptotic cell death in isolated working rat hearts. PMID- 12575924 TI - Vaginal bacterial flora activates rat peritoneal mast cells. AB - Sixteen strains of physiological and pathological vaginal bacteria were tested for their ability to secrete histamine from rat peritoneal mast cells in vitro. We noticed that Mycoplasma hominis-induced histamine release was very high (up to 53.6%). The stimulation of rat mast cells with Staphylococccus cohnii, Staphylococcus coagulase(-) (two strains), Ureaplasma urealyticum, Peptostreptococcus spp., Bacteroides capillosus, Staphylococcus aureus and Streptococcus agalactiae resulted in lower but significant histamine secretion (11.2%-17.5%). Other bacteria strains (Staphylococcus epidermidids, Enterococcus faecalis, Escherichia coli, Actinomyces naeslundii (two strains) and Lactobacillus fermentum (two strains) caused very low (4.2% - 8.8%) histamine release. PMID- 12575925 TI - Evidence for a T-lymphocyte dependent mechanism involving proinflammatory cytokines in anticonvulsant hypersensitivity syndrome: a case report. AB - The anticonvulsant drug hypersensitivity syndrome (AHS) is a potentially fatal drug reaction associated with aromatic antiepileptic drugs, the pathogenic mechanisms of which are still unclear. We studied a 2-year-old female affected with AHS after phenobarbital treatment. In vitro experiments showed a T lymphocyte response to the drug and an increased, dose-related, IL-6 production after drug incubation. These findings suggest that AHS may be caused by a T lymphocyte dependent mechanism, involving pro-inflammatory cytokines. PMID- 12575926 TI - Interleukin 10 and transforming growth factor beta 1 in hepatic fibrosis. PMID- 12575927 TI - Prevalence of IgG antibodies against Borrelia Burgdorferi s.l. and Ehrlichia Phagocytophila in sera of patients presenting symptoms of Lyme disease in a central region of Italy. AB - The aim of this study was to evaluate the prevalence (seroprevalence) of antibodies against Borrelia burgdorferi and Ehrlichia phagocytophila among patients resident in Lazio, a region of central Italy. Of a sample of 1,050 patients, which presented clinical manifestations related to Lyme disease, 34 (3.2%) were Borrelia-seropositive (Lyme index value >/= 1.2). The sera of 25 out of the 34 patients that were Borrelia-positive were also analysed for the presence of antibodies against E. phagocytophila and 3 (12%) were found Ehrlichia positive (titres >1:64). No Ehrlichia positive samples were found among sera of 250 Borrelia-negative patients. Since both B. burgdorferi s.l. and Ehrlichia species share the same tick vector (Ixodes ricinus), our results indicate that concurrent transmission of these microbial pathogens might have been occurred among the patients included in this study. PMID- 12575928 TI - Mast cell deficient W/W(v) mice lack stress-induced increase in serum IL-6 levels, as well as in peripheral CRH and vascular permeability, a model of rheumatoid arthritis. AB - Corticotropin releasing hormone (CRH) and interleukin-6 (IL-6) are implicated in inflammatory diseases triggered by stress. Acute restraint stress increases serum IL-6 in the blood, but its source is not known. Our current study was carried out in order to determine the contribution of mast cells to stress-induced IL-6 release and to investigate skin CRH and vascular permeability in mice. W/W(v) mast cell deficient and their wild type control +/+ mice were stressed in a plexiglass restraint chamber for 60 or 120 min. Serum corticosterone and IL-6 levels were measured. Other mice were injected with (99)Tchnetium gluceptate ((99)Tc) and its extravastion, indicating vascular permeability, was determined along with CRH levels in the skin and knee joints. Acute stress increased serum IL-6 in mice, but was greatly inhibited in W/W(v) mast cell deficient mice. Vascular permeability to (99)Tc, as well as local CRH levels, were also increased by stress, but not in W/W(v) mice. Findings from our current study suggest a link between mast cells and stress-related skin and joint inflammation and may explain initial events in psoriatic and rheumatoid arthritis. PMID- 12575929 TI - Picking the lock with a hairpin. Did HIV-1 learn the art from chemokines? PMID- 12575930 TI - Switching off the switch. PMID- 12575931 TI - An intramolecular spin of the LDL receptor beta propeller. AB - Study of the LDL receptor as a model system has led to insights into general principles underlying receptor-mediated endocytosis of bound ligands. The recently published structure of the entire LDL receptor ectodomain, determined at pH 5.3, now suggests an elegant model to explain how lipoprotein ligands are released from the receptor by exposure to the low-pH environment of the endosome. PMID- 12575932 TI - A primordial structure underlying amyloid. AB - The parallel beta helix is a good model of the "primordial" structure underlying amyloid: incompatible sequences can spill out as loops between the helical turns, and the cross-section and internal hydration can vary according to the nature of the amyloid-forming sequence. PMID- 12575933 TI - Dehydration converts DsbG crystal diffraction from low to high resolution. AB - Diffraction quality crystals are essential for crystallographic studies of protein structure, and the production of poorly diffracting crystals is often regarded as a dead end in the process. Here we show a dramatic improvement of poorly diffracting DsbG crystals allowing high-resolution diffraction data measurement. Before dehydration, the crystals are fragile and the diffraction pattern is streaky, extending to 10 A resolution. After dehydration, there is a spectacular improvement, with the diffraction pattern extending to 2 A resolution. This and other recent results show that dehydration is a simple, rapid, and inexpensive approach to convert poor quality crystals into diffraction quality crystals. PMID- 12575934 TI - Catalysis, specificity, and ACP docking site of Streptomyces coelicolor malonyl CoA:ACP transacylase. AB - Malonyl-CoA:ACP transacylase (MAT), the fabD gene product of Streptomyces coelicolor A3(2), participates in both fatty acid and polyketide synthesis pathways, transferring malonyl groups that are used as extender units in chain growth from malonyl-CoA to pathway-specific acyl carrier proteins (ACPs). Here, the 2.0 A structure reveals an invariant arginine bound to an acetate that mimics the malonyl carboxylate and helps define the extender unit binding site. Catalysis may only occur when the oxyanion hole is formed through substrate binding, preventing hydrolysis of the acyl-enzyme intermediate. Macromolecular docking simulations with actinorhodin ACP suggest that the majority of the ACP docking surface is formed by a helical flap. These results should help to engineer polyketide synthases (PKSs) that produce novel polyketides. PMID- 12575935 TI - Solution structure of the MAPK phosphatase PAC-1 catalytic domain. Insights into substrate-induced enzymatic activation of MKP. AB - Inactivation of mitogen-activated protein kinases (MAPKs) by MAPK phosphatases (MKPs) is accomplished via substrate-induced activation of the latter enzymes; however, the structural basis for the underlying mechanism remains elusive. Here, we report the three-dimensional solution structure of the C-terminal phosphatase domain of the prototypical MKP PAC-1, determined when bound to phosphate. Structural and biochemical analyses reveal unique active site geometry of the enzyme important for binding to phosphorylated threonine and tyrosine of MAPK ERK2. Our study further demonstrates that the dynamic interaction between the N terminal kinase binding domain and the C-terminal phosphatase domain of an MKP is directly coupled to MAPK-induced conformational change of the phosphatase active site, which is essential for eliciting its full enzymatic activity. PMID- 12575936 TI - A novel protein fold and extreme domain swapping in the dimeric TorD chaperone from Shewanella massilia. AB - TorD is the cytoplasmic chaperone involved in the maturation of the molybdoenzyme TorA prior to the translocation of the folded protein into the periplasm. The X ray structure at 2.4 A resolution of the TorD dimer reveals extreme domain swapping between the two subunits. The all-helical architecture of the globular domains within the intertwined molecular dimer shows no similarity with known protein structures. According to sequence similarities, this new fold probably represents the architecture of the chaperones associated with the bacterial DMSO/TMAO reductases and also that of proteins of yet unknown functions. The occurrence of multiple oligomeric forms and the chaperone activity of both monomeric and dimeric TorD raise questions about the possible biological role of domain swapping in this protein. PMID- 12575937 TI - The crystal structure of Drosophila NLP-core provides insight into pentamer formation and histone binding. AB - The nucleoplasmin-like protein from Drosophila (dNLP) functions as a chaperone for core histones and may remodel chromatin in embryos. We now report the crystal structure of a dNLP-core pentamer at 1.5 A resolution. The monomer has an eight stranded, beta barrel topology that is similar to nucleoplasmin (Np). However, a signature beta hairpin is tucked in along the lateral surface of the dNLP-core pentamer, while it extends outward in the Np-core decamer. Drosophila NLP and Np both assemble histone octamers. This process may require each chaperone to form a decamer, which would create symmetric binding sites for the histones. Conformational differences between dNLP and Np may reflect their different oligomeric states, while a conserved, nonpolar subunit interface may allow conformational plasticity during histone binding. PMID- 12575938 TI - ATP-mediated conformational changes in the RecA filament. AB - The crystal structure of the E. coli RecA protein was solved more than 10 years ago, but it has provided limited insight into the mechanism of homologous genetic recombination. Using electron microscopy, we have reconstructed five different states of RecA-DNA filaments. The C-terminal lobe of the RecA protein is modulated by the state of the distantly bound nucleotide, and this allosteric coupling can explain how mutations and truncations of this C-terminal lobe enhance RecA's activity. A model generated from these reconstructions shows that the nucleotide binding core is substantially rotated from its position in the RecA crystal filament, resulting in ATP binding between subunits. This simple rotation can explain the large cooperativity in ATP hydrolysis observed for RecA DNA filaments. PMID- 12575939 TI - Novel fold revealed by the structure of a FAS1 domain pair from the insect cell adhesion molecule fasciclin I. AB - Fasciclin I is an insect neural cell adhesion molecule consisting of four FAS1 domains, homologs of which are present in many bacterial, plant, and animal proteins. The crystal structure of FAS1 domains 3 and 4 of Drosophila fasciclin I reveals a novel domain fold, consisting of a seven-stranded beta wedge and a number of alpha helices. The two domains are arranged in a linear fashion and interact through a substantial polar interface. Missense mutations in the FAS1 domains of the human protein betaig-h3 cause corneal dystrophies. Many mutations alter highly conserved core residues, but the two most common mutations, affecting Arg-124 and Arg-555, map to exposed alpha-helical regions, suggesting reduced protein solubility as the disease mechanism. PMID- 12575940 TI - Crystal structure of protein C inhibitor provides insights into hormone binding and heparin activation. AB - Protein C inhibitor (PCI) is a member of the serpin family that has many biological functions. In blood it acts as a procoagulant, and, in the seminal vesicles, it is required for spermatogenesis. The activity of PCI is affected by heparin binding in a manner unique among the heparin binding serpins, and, in addition, PCI binds hydrophobic hormones with apparent specificity for retinoids. Here we present the 2.4 A crystallographic structure of reactive center loop (RCL) cleaved PCI. A striking feature of the structure is a two-turn N-terminal shortening of helix A, which creates a large hydrophobic pocket that docking studies indicate to be the retinoid binding site. On the basis of surface electrostatic properties, a novel mechanism for heparin activation is proposed. PMID- 12575941 TI - Specific radiation damage can be used to solve macromolecular crystal structures. AB - The use of third generation synchrotron sources has led to renewed concern about the effect of ionizing radiation on crystalline biological samples. In general, the problem is seen as one to be avoided. However, in this paper, it is shown that, far from being a hindrance to successful structure determination, radiation damage provides an opportunity for phasing macromolecular structures. This is successfully demonstrated for both a protein and an oligonucleotide, by way of which complete models were built automatically. The possibility that, through the exploitation of radiation damage, the phase problem could become less of a barrier to macromolecular crystal structure determination is discussed. PMID- 12575942 TI - Alternative conformations of HIV-1 V3 loops mimic beta hairpins in chemokines, suggesting a mechanism for coreceptor selectivity. AB - The V3 loop of the HIV-1 envelope glycoprotein gp120 is involved in binding to the CCR5 and CXCR4 coreceptors. The structure of an HIV-1(MN) V3 peptide bound to the Fv of the broadly neutralizing human monoclonal antibody 447-52D was solved by NMR and found to be a beta hairpin. This structure of V3(MN) was found to have conformation and sequence similarities to beta hairpins in CD8 and CCR5 ligands MIP-1alpha, MIP-1beta, and RANTES and differed from the beta hairpin of a V3(IIIB) peptide bound to the strain-specific murine anti-gp120(IIIB) antibody 0.5beta. In contrast to the structure of the bound V3(MN) peptide, the V3(IIIB) peptide resembles a beta hairpin in SDF-1, a CXCR4 ligand. These data suggest that the 447-52D-bound V3(MN) and the 0.5beta-bound V3(IIIB) structures represent alternative V3 conformations responsible for selective interactions with CCR5 and CXCR4, respectively. PMID- 12575943 TI - Conservation and specialization. The role of the exocyst in neuronal exocytosis. AB - The exocyst is an octameric complex mediating vesicle targeting and tethering at the plasma membrane for exocytosis. The role of exocyst in nervous system is unclear. In this issue of Neuron, Murthy et al. provide important insights: defects in the exocyst inhibit neurite outgrowth and neuromuscular junction formation; however, synaptic transmission persists. PMID- 12575944 TI - It takes two to tango, but three to ISA. AB - Rapidly inactivating A-type potassium channels are important determinants of firing frequency in many excitable cells. Nadal et al. (in this issue of Neuron) purified A-type potassium (I(SA)) channels from rat cerebellum and identified a novel beta subunit. This protein, DPPX, associates with the pore-forming subunits and endows previously elusive kinetic properties on A-type channels formed from cloned subunits. PMID- 12575945 TI - Hippocampus and memory. Can we have our place and fear it too? AB - Theories of hippocampal function are often split into cognitive map theories and relational/episodic memory theories. In this issue of Neuron, show that hippocampal cells respond to the combination of spatial location and conditioned stimuli in a nonspatial fear conditioning task, suggesting a potential bridge between the competing theories of hippocampal function. PMID- 12575946 TI - GABA generates excitement. AB - In the CNS, gamma-aminobutyric acid (GABA) acts as an inhibitory transmitter via ligand-gated GABA(A) receptor channels and G protein-coupled GABA(B) receptors. Both of these receptor types mediate inhibitory postsynaptic transmission throughout the nervous system. For GABA(A) receptors, this inhibitory action is associated with a hyperpolarization due to an increase in conductance to chloride ions. Previous studies show that GABA(A) receptor activation in neonatal neurons and spinal cord neurons can be excitatory. Two papers recently appeared that clearly demonstrate that GABA can have a depolarizing and excitatory action in mature cortical neurons. Here we discuss the evolving story on chloride ion homeostasis in CNS neurons and its role in the bipolar life of the GABA(A) receptor. PMID- 12575947 TI - Synaptic ribbon. Conveyor belt or safety belt? AB - The synaptic ribbon in neurons that release transmitter via graded potentials has been considered as a conveyor belt that actively moves vesicles toward their release sites. But evidence has accumulated to the contrary, and it now seems plausible that the ribbon serves instead as a safety belt to tether vesicles stably in mutual contact and thus facilitate multivesicular release by compound exocytosis. PMID- 12575948 TI - SCA7 knockin mice model human SCA7 and reveal gradual accumulation of mutant ataxin-7 in neurons and abnormalities in short-term plasticity. AB - We targeted 266 CAG repeats (a number that causes infantile-onset disease) into the mouse Sca7 locus to generate an authentic model of spinocerebellar ataxia type 7 (SCA7). These mice reproduced features of infantile SCA7 (ataxia, visual impairments, and premature death) and showed impaired short-term synaptic potentiation; downregulation of photoreceptor-specific genes, despite apparently normal CRX activity, led to shortening of photoreceptor outer segments. Wild-type ataxin-7 was barely detectable, as was mutant ataxin-7 in young animals; with increasing age, however, ataxin-7 staining became more pronounced. Neurons that appeared most vulnerable had relatively high levels of mutant ataxin-7; it is interesting, however, that marked dysfunction occurred in these neurons weeks prior to the appearance of nuclear inclusions. These data demonstrate that glutamine expansion stabilizes mutant ataxin-7, provide an explanation for selective neuronal vulnerability, and show that mutant ataxin-7 impairs posttetanic potentiation (PTP). PMID- 12575949 TI - Integrity of developing spinal motor columns is regulated by neural crest derivatives at motor exit points. AB - Spinal motor neurons must extend their axons into the periphery through motor exit points (MEPs), but their cell bodies remain within spinal motor columns. It is not known how this partitioning is established in development. We show here that motor neuron somata are confined to the CNS by interactions with a neural crest subpopulation, boundary cap (BC) cells that prefigure the sites of spinal MEPs. Elimination of BC cells by surgical or targeted genetic ablation does not perturb motor axon outgrowth but results in motor neuron somata migrating out of the spinal cord by translocating along their axons. Heterologous neural crest grafts in crest-ablated embryos stop motor neuron emigration. Thus, before the formation of a mature transitional zone at the MEP, BC cells maintain a cell tight boundary that allows motor axons to cross but blocks neuron migration. PMID- 12575950 TI - RNA cargoes associating with FMRP reveal deficits in cellular functioning in Fmr1 null mice. AB - The Fragile X mental retardation-1 (Fmr1) gene encodes a multifunctional protein, FMRP, with intrinsic RNA binding activity. We have developed an approach, antibody-positioned RNA amplification (APRA), to identify the RNA cargoes associated with the in vivo configured FMRP messenger ribonucleoprotein (mRNP) complex. Using APRA as a primary screen, putative FMRP RNA cargoes were assayed for their ability to bind directly to FMRP using traditional methods of assessing RNA-protein interactions, including UV-crosslinking and filter binding assays. Approximately 60% of the APRA-defined mRNAs directly associate with FMRP. By examining a subset of these mRNAs and their encoded proteins in brain tissue from Fmr1 knockout mice, we have observed that some of these cargoes as well as the proteins they encode show discrete changes in abundance and/or differential subcellular distribution. These data are consistent with spatially selective regulation of multiple biological pathways by FMRP. PMID- 12575951 TI - Mutations in the exocyst component Sec5 disrupt neuronal membrane traffic, but neurotransmitter release persists. AB - The exocyst (Sec6/8) complex is necessary for secretion in yeast and has been postulated to establish polarity by directing vesicle fusion to specific sites along the plasma membrane. The complex may also function in the nervous system, but its precise role is unknown. We have investigated exocyst function in Drosophila with mutations in one member of the complex, sec5. Null alleles die as growth-arrested larvae, whose neuromuscular junctions fail to expand. In culture, neurite outgrowth fails in sec5 mutants once maternal Sec5 is exhausted. Using a trafficking assay, we found impairments in the membrane addition of newly synthesized proteins. In contrast, synaptic vesicle fusion was not impaired. Thus, Sec5 differentiates between two forms of vesicle trafficking: trafficking for cell growth and membrane protein insertion depend on sec5, whereas transmitter secretion does not. In this regard, sec5 differs from the homologs of other yeast exocytosis genes that are required for both neuronal trafficking pathways. PMID- 12575953 TI - Bidirectional changes in spatial dendritic integration accompanying long-term synaptic modifications. AB - Information processing in the neuron requires spatial summation of synaptic inputs at the dendrite. In CA1 pyramidal neurons of the hippocampus, a brief period of correlated pre- and postsynaptic activity, which induces long-term potentiation (LTP) or long-term depression (LTD), results in a persistent increase or decrease in the linearity of spatial summation, respectively. Such bidirectional modification of the summation property is specific to the modified input and reflects localized dendritic changes involving I(h) channels and NMDA receptors. Thus, correlated pre- and postsynaptic activity alters not only the strength of the activated input but also its dendritic integration with other inputs. PMID- 12575952 TI - The CD26-related dipeptidyl aminopeptidase-like protein DPPX is a critical component of neuronal A-type K+ channels. AB - Subthreshold-activating somatodendritic A-type potassium channels have fundamental roles in neuronal signaling and plasticity which depend on their unique cellular localization, voltage dependence, and kinetic properties. Some of the components of A-type K(+) channels have been identified; however, these do not reproduce the properties of the native channels, indicating that key molecular factors have yet to be unveiled. We purified A-type K(+) channel complexes from rat brain membranes and found that DPPX, a protein of unknown function that is structurally related to the dipeptidyl aminopeptidase and cell adhesion protein CD26, is a novel component of A-type K(+) channels. DPPX associates with the channels' pore-forming subunits, facilitates their trafficking and membrane targeting, reconstitutes the properties of the native channels in heterologous expression systems, and is coexpressed with the pore forming subunits in the somatodendritic compartment of CNS neurons. PMID- 12575954 TI - Activation of a tyrosine kinase-MAPK cascade enhances the induction of long-term synaptic facilitation and long-term memory in Aplysia. AB - Tyrosine kinases have been implicated in cellular processes thought to underlie learning and memory. Here we show that tyrosine kinases play a direct role in long-term synaptic facilitation (LTF) and long-term memory (LTM) for sensitization in Aplysia. Tyrosine kinase activity is required for serotonin induced LTF of sensorimotor (SN-MN) synapses, and enhancement of endogenous tyrosine kinase activity facilitates the induction of LTF. These effects are mediated, at least in part, through mitogen-activated protein kinase (MAPK) activation and are blocked by transcriptional and translational inhibitors. Moreover, brain-derived neurotrophic factor (BDNF) also enhances the induction of LTF in a MAPK-dependent fashion. Finally, activation of endogenous tyrosine kinases enhances the induction of long-term memory for sensitization, and this enhancement also requires MAPK activation. Thus, tyrosine kinases, acting through MAPK, play a pivotal role in LTF and LTM formation. PMID- 12575955 TI - Hippocampal place cells acquire location-specific responses to the conditioned stimulus during auditory fear conditioning. AB - We recorded neurons from the hippocampus of freely behaving rats during an auditory fear conditioning task. Rats received either paired or unpaired presentations of an auditory conditioned stimulus (CS) and an electric shock unconditioned stimulus (US). Hippocampal neurons (place and theta cells) acquired responses to the auditory CS in the paired but not in the unpaired group. After CS-US pairing, rhythmic firing of theta cells became synchronized to the onset of the CS. Conditioned responses of place cells were gated by their location specific firing, so that after CS-US pairing, place cells responded to the CS only when the rat was within the cell's place field. These findings may help to elucidate how the hippocampus contributes to context-specific memory formation during associative learning. PMID- 12575956 TI - Multineuronal firing patterns in the signal from eye to brain. AB - Population codes in the brain have generally been characterized by recording responses from one neuron at a time. This approach will miss codes that rely on concerted patterns of action potentials from many cells. Here we analyze visual signaling in populations of ganglion cells recorded from the isolated salamander retina. These neurons tend to fire synchronously far more frequently than expected by chance. We present an efficient algorithm to identify what groups of cells cooperate in this way. Such groups can include up to seven or more neurons and may account for more than 50% of all the spikes recorded from the retina. These firing patterns represent specific messages about the visual stimulus that differ significantly from what one would derive by single-cell analysis. PMID- 12575957 TI - Adaptive coincidence detection and dynamic gain control in visual cortical neurons in vivo. AB - Several theories have proposed a functional role for response synchronization in sensory perception. Critics of these theories have argued that selective synchronization is physiologically implausible when cortical networks operate at high levels of activity. Using intracellular recordings from visual cortex in vivo, in combination with numerical simulations, we find dynamic changes in spike threshold that reduce cellular sensitivity to slow depolarizations and concurrently increase the relative sensitivity to rapid depolarizations. Consistent with this, we find that spike activity and high-frequency fluctuations in membrane potential are closely correlated and that both are more tightly tuned for stimulus orientation than the mean membrane potential. These findings suggest that under high-input conditions the spike-generating mechanism adaptively enhances the sensitivity to synchronous inputs while simultaneously decreasing the sensitivity to temporally uncorrelated inputs. PMID- 12575958 TI - Two-dimensional substructure of stereo and motion interactions in macaque visual cortex. AB - The analysis of object motion and stereoscopic depth are important tasks that are begun at early stages of the primate visual system. Using sparse white noise, we mapped the receptive field substructure of motion and disparity interactions in neurons in V1 and MT of alert monkeys. Interactions in both regions revealed subunits similar in structure to V1 simple cells. For both motion and stereo, the scale and shape of the receptive field substructure could be predicted from conventional tuning for bars or dot-field stimuli, indicating that the small scale interactions were repeated across the receptive fields. We also found neurons in V1 and in MT that were tuned to combinations of spatial and temporal binocular disparities, suggesting a possible neural substrate for the perceptual Pulfrich phenomenon. Our observations constrain computational and developmental models of motion-stereo integration. PMID- 12575959 TI - Neural correlates of visual localization and perisaccadic mislocalization. AB - While reading this text, your eyes jump from word to word. Yet you are unaware of the motion this causes on your retina; the brain somehow compensates for these displacements and creates a stable percept of the world. This compensation is not perfect; perisaccadically, perceptual space is distorted. We show that this distortion can be traced to a representation of retinal position in the medial temporal and medial superior temporal areas. These cells accurately represent retinal position during fixation, but perisaccadically, the same cells distort the representation of space. The time course and magnitude of this distortion are similar to the mislocalization found psychophysically in humans. This challenges the assumption in many psychophysical studies that the perisaccadic retinal position signal is veridical. PMID- 12575960 TI - Symptomatic myocardial bridges: overview of ischemic mechanisms and current diagnostic and treatment strategies. AB - This review article focuses on the morphological and functional alterations that characterize patients with myocardial bridges (MB) as well as the currently available diagnostic and treatment strategies. Because of incomplete understanding of the pathophysiology of MB, their clinical significance has been the subject of debate for the last quarter century. Investigational tools now available in the cardiac catheterization laboratory have helped clarify why symptoms and signs of ischemia can occur in such patients, especially when the only angiographic finding appears to be systolic compression or milking effect of a coronary vessel. Quantitative coronary angiography and intravascular ultrasound (IVUS) clearly demonstrate that the phasic systolic vessel compression visualized on the angiogram is coupled with a persistent diastolic diameter reduction. Intracoronary Doppler reveals increased flow velocities, retrograde systolic flow, and reduced coronary flow reserve. The clinical diagnosis can be established by significant percent lumen diameter and area narrowing, increased flow velocity, and by characteristic patterns such as the "half moon" phenomenon on IVUS and the early diastolic "finger tip" phenomenon on intracoronary Doppler. Successful medical, interventional, or surgical therapy leads not only to marked improvement or normalization of these alterations but also relief of angina and ischemia. PMID- 12575961 TI - Temporal increases in plasma markers of oxidized low-density lipoprotein strongly reflect the presence of acute coronary syndromes. AB - OBJECTIVES: This study was conducted to test the hypothesis that plasma markers of oxidized low-density lipoprotein (OxLDL) reflect acute coronary syndromes (ACS). BACKGROUND: Oxidized LDL contributes to the pathogenesis of atherosclerosis, but its role in ACS is not established. METHODS: Serial plasma samples were prospectively obtained from patients with an acute myocardial infarction (MI) (n = 8), unstable angina (UA) (n = 15), stable coronary artery disease (CAD) (n = 17), angiographically normal coronary arteries (n = 8), and from healthy subjects (n = 18), at entry into the study, hospital discharge (MI group only), and at 30, 120, and 210 days. Chemiluminescent enzyme-linked immunosorbent assay was used to quantitate plasma levels of: 1) immunoglobulin (Ig)M and IgG OxLDL autoantibody titers (presented as a mean OxLDL autoantibody titer by averaging the results of four distinct epitopes); 2) LDL-autoantibody immune complexes (LDL-IC); and 3) minimally OxLDL measured by antibody E06 (OxLDL E06), as determined by the content of oxidized phospholipids (OxPL) per apolipoprotein B-100. RESULTS: Baseline OxLDL IgG autoantibody levels were higher in the MI group (p < 0.0001). At 30-day follow-up, the mean IgM OxLDL titers increased by 48% (p < 0.001) and 20% (p < 0.001), and IgM LDL-IC increased by 60% (p < 0.01) and 26% (p < 0.01) in the MI and UA groups, respectively. The OxLDL E06 levels increased by 54% (p < 0.01) in the MI group at hospital discharge and by 36% at 30 days. No significant changes in any OxLDL markers were noted in the other groups. The OxLDL-E06 levels strongly paralleled the acute rise in lipoprotein(a), or Lp(a), in the MI group, suggesting that toxic OxPL are preferentially bound to Lp(a). Oxidized LDL-E06 also correlated extremely well with Lp(a) in the entire cohort of patients (r = 0.91, p < 0.0001). CONCLUSIONS: Circulating OxLDL-specific markers strongly reflect the presence of ACS, implying immune awareness to newly exposed oxidation-specific epitopes and possible release of OxLDL in the circulation. The OxLDL-E06 measurements provide novel insights into plaque rupture and the potential atherogenicity of Lp(a). PMID- 12575962 TI - Troponin T and quantitative ST-segment depression offer complementary prognostic information in the risk stratification of acute coronary syndrome patients. AB - OBJECTIVES: Our primary objective was to examine the prognostic relationship between baseline quantitative ST-segment depression (ST) and cardiac troponin T (cTnT) elevation. The secondary objectives were to: 1) examine whether ST provided additional insight into therapeutic efficacy of glycoprotein IIb/IIIa therapy similar to that demonstrated by cTnT; and 2) explore whether the time to evaluation impacted on each marker's relative prognostic utility. BACKGROUND: The relationship between the baseline electrocardiogram (ECG) and cTnT measurements in risk-stratifying patients presenting with acute coronary syndromes (ACS) has not been evaluated comprehensively. METHODS: The study population consisted of 959 patients enrolled in the cTnT substudy of the Platelet IIb/IIIa Antagonism for the Reduction of Acute coronary syndrome events in a Global Organization Network (PARAGON)-B trial. Patients were classified as having no ST (n = 387), 1 mm ST (n = 433), and ST > or =2 mm (n = 139). Forty-percent (n = 381) were classified as cTnT-positive based on a definition of > or =0.1 ng/ml. RESULTS: Six-month death/(re)myocardial infarction rates were 8.4% among cTnT-negative patients with no ST and 26.8% among cTnT-positive patients with ST > or =2 mm. On ECGs done after 6 h of symptom onset, ST > or =2 mm was associated with higher risk compared to its presence on ECGs done earlier (odds ratio [OR] 7.3 vs. 2.1). In contrast, the presence of elevated cTnT within 6 h of symptom was associated with a higher risk of adverse events compared with elevations after 6 h (OR 2.4 vs. 1.5). CONCLUSIONS: Quantitative ST and cTnT status are complementary in assessing risk among ACS patients and both should be employed to determine prognosis and assist in medical decision making. PMID- 12575963 TI - Severe renal dysfunction complicating cardiogenic shock is not a contraindication to mechanical support as a bridge to cardiac transplantation. AB - OBJECTIVES: This study investigated outcomes in patients with cardiogenic shock and severe renal dysfunction treated with ventricular assist devices (VAD) as a bridge to cardiac transplantation. BACKGROUND: Previous reports have documented poor survival in patients with cardiogenic shock and severe renal dysfunction treated with VAD. METHODS: We surveyed 215 consecutive patients who received a VAD from 1992 to 2000 and selected patients who had a serum creatinine > or =3.0 mg/dl at the time of VAD placement. Demographic, laboratory, and clinical outcome data were collected. RESULTS: Eighteen patients met the inclusion criteria. Mean serum creatinine at the time of VAD placement was 4.0 +/- 0.7 mg/dl (range 3.0 to 5.2 mg/dl). Seven patients required temporary renal support with continuous venovenous hemodialysis (CVVHD). Eleven patients underwent cardiac transplantation. At six months post-transplantation, mean serum creatinine was 2.0 +/- 0.6 mg/dl (range 1.3 to 3.5 mg/dl). None of the transplanted patients required subsequent renal support. Seven patients died with a VAD before transplantation. Three died early (<1 month) after VAD placement, and all three required CVVHD until death. Four patients survived for >1 month after VAD placement; all four had resolution of renal dysfunction with mean serum creatinine of 1.9 +/- 1.2 mg/dl (range 0.8 to 3.6 mg/dl) without the need for renal support. Overall 30-day and six-month survival after VAD placement, survival to transplantation, and survival one year post-transplantation were similar to patients without severe renal dysfunction. CONCLUSIONS: Contemporary use of VAD leads to resolution of severe renal dysfunction in most cardiogenic shock patients and comparable long-term outcomes to patients without renal dysfunction. PMID- 12575964 TI - Relationship between heparin anticoagulation and clinical outcomes in coronary stent intervention: observations from the ESPRIT trial. AB - OBJECTIVES: We evaluated the relationship between the degree of heparin anticoagulation and clinical efficacy and bleeding in patients undergoing contemporary percutaneous coronary intervention (PCI) with stent implantation. BACKGROUND: Despite universal acceptance of heparin anticoagulation as a standard of care in PCI, considerable controversy still exists regarding the appropriate dosing of heparin. METHODS: The study population (n = 2,064) comprised all patients enrolled in the Enhanced Suppression of the Platelet IIb/IIIa Receptor with Integrilin Therapy (ESPRIT) trial. The index activated clotting time (ACT) was defined as the ACT measured after the last heparin dose and before first device activation and was correlated with outcome and bleeding events. RESULTS: No association was observed between decreasing ACT levels and the rate of ischemic events in the treatment or placebo arms. The incidence of the primary composite end point (death, myocardial infarction, urgent target vessel revascularization, and thrombotic bailout glycoprotein IIb/IIIa inhibitor therapy at 48 h) was actually lowest in the lowest ACT tertile for both the placebo (10.0%) and treatment groups (6.1%). When analyzed by tertile, major bleeding rates did not increase in the lowest ACT tertile in patients given placebo (0.6%) versus those receiving eptifibatide (0.7%). Major bleeding rates increased as the ACT increased in the eptifibatide-treated patients. CONCLUSIONS: Ischemic end points in patients undergoing contemporary PCI with stent placement do not increase by decreasing ACT levels, at least to a level of 200 s. Bleeding events do increase with increasing ACT levels and are enhanced with eptifibatide treatment. An ACT of 200 to 250 s is reasonable in terms of efficacy and safety with the use of contemporary technology and pharmacotherapy. PMID- 12575965 TI - The activated clotting time can be used to monitor the low molecular weight heparin dalteparin after intravenous administration. AB - OBJECTIVES: This study was designed to compare the dose response of dalteparin versus unfractionated heparin (UFH) on the activated clotting time (ACT), and to determine whether the ACT can be used to monitor intravenous (IV) dalteparin during percutaneous coronary intervention (PCI). BACKGROUND: The use of low molecular weight heparin (LMWH) during PCI has been limited by the presumed inability to monitor its anticoagulant effect using bedside assays. METHODS: This study was performed in three phases. In vitro, ACTs were measured on volunteer (n = 10) blood samples spiked with increasing concentrations of dalteparin or UFH. To extend these observations in vivo, ACTs were then measured in patients (n = 15) who were sequentially treated with IV dalteparin and then UFH. Finally, a larger monitoring study was undertaken involving patients (n = 110) who received dalteparin 60 or 80 international U (IU)/kg alone or followed by abciximab. We measured ACT (Hemochron), activated partial thromboplastin time (aPTT), plasma anti-Xa and anti-IIa levels, tissue factor pathway inhibitor (TFPI) concentration, and plasma dalteparin concentration. RESULTS: Dalteparin induced a significant rise in the ACT with a smaller degree of variance as compared to UFH. Five min after administration of IV dalteparin 80 IU/kg the ACT increased from 125 s (122 s, 129 s) to 184 s (176 s, 191 s) (p < 0.001). The aPTT, anti-Xa and anti-IIa activities, and TFPI concentration also demonstrated significant increases following IV dalteparin. CONCLUSIONS: The ACT and aPTT are sensitive to IV dalteparin at clinically relevant doses. These data suggest that the ACT may be useful in monitoring the anticoagulant effect of intravenously administered dalteparin during PCI. PMID- 12575966 TI - Thermal heterogeneity in stable human coronary atherosclerotic plaques is underestimated in vivo: the "cooling effect" of blood flow. AB - OBJECTIVES: This study investigated whether temperature measurements are influenced by blood flow. BACKGROUND: Previous ex vivo studies showed marked thermal heterogeneity in atheromatic plaques. In stable lesions, however, trivial in vivo temperature variations are recorded, perhaps due to the "cooling effect" of blood flow. METHODS: Eighteen patients with effort angina were studied. Coronary flow velocity was continuously recorded; over another guidewire, temperature measurements were performed at the proximal vessel wall and at the lesion before, during, and after complete interruption of blood flow by inflation of a balloon. The DeltaTp was assigned as the difference between the proximal vessel wall temperature and the maximal temperature during and after balloon inflation. The DeltaTl was assigned as the difference between the atherosclerotic plaque and the proximal vessel wall. RESULTS: The procedure was not complicated. During and after complete interruption of flow, DeltaTp was 0.012 +/- 0.01 degrees C and -0.006 +/- -0.01 degrees C (p < 0.001), respectively. The DeltaTl was 0.08 +/- 0.04 degrees C at baseline and went to 0.18 +/- 0.05 degrees C (60.5 +/- 14.1% increase) during and 0.08 +/- 0.04 degrees C after flow interruption (p < 0.001). The DeltaTl was greater than DeltaTp during and after impairment of flow (p < 0.001). A correlation between the baseline average peak velocity and DeltaTl during flow interruption was found (R = 0.57, p = 0.01). In seven patients thermal heterogeneity was not detected at baseline, and during balloon inflation DeltaTl increased by 76.0 +/- 8.4%. CONCLUSIONS: Thermal heterogeneity is underestimated in atherosclerotic plaques in patients with effort angina. Potential in vivo underestimation of heat production locally in human atherosclerotic is due to the "cooling effect" of coronary blood flow. PMID- 12575967 TI - Late acute thrombotic occlusion after endovascular brachytherapy and stenting of femoropopliteal arteries. AB - OBJECTIVES: The aim of this article is to underline the importance of this complication after endovascular brachytherapy (EVBT) and intravascular stenting of the femoropopliteal arteries occurring in a running randomized trial. BACKGROUND: Endovascular brachytherapy has been proposed as a promising treatment modality to reduce restenosis after angioplasty. However, the phenomenon of late acute thrombotic occlusion (LATO) in patients receiving EVBT after stenting is of major concern. METHODS: In an ongoing prospective multicenter trial, patients were randomized to undergo EVBT (iridium 192; 14 Gy at a depth of the radius of the vessel +2 mm) after percutaneous recanalization of femoropopliteal obstructions. Of the 204 patients who completed the six months follow-up, 94 were randomized to EVBT. RESULTS: Late acute thrombotic occlusion occurred exclusively in 6 of 22 patients (27%) receiving EVBT after intravascular stenting and always in concomitance with reduction of antithrombotic drug prevention (clopidogrel). Conversely, none of the 13 patients with stents and without EVBT (0%; p < 0.05) and none of the 72 patients (0%; p < 0.01) undergoing EVBT after simple balloon angioplasty presented LATO. CONCLUSIONS: Late thrombotic occlusion occurs not only in patients undergoing EVBT after percutaneous coronary recanalization but also after stenting of the femoropopliteal arteries and may compromise the benefits of endovascular radiation. The fact that all our cases with LATO occurred concomitantly with stopping clopidogrel may indicate a possible rebound mechanism. An intensive and prolonged antithrombotic prevention is probably indicated in these patients. PMID- 12575968 TI - Hypoestrogenemia of hypothalamic origin and coronary artery disease in premenopausal women: a report from the NHLBI-sponsored WISE study. AB - OBJECTIVES: We sought to evaluate hypoestrogenemia of hypothalamic origin and its association with angiographic coronary artery disease (CAD) in premenopausal women. BACKGROUND: Coronary artery disease in premenopausal women appears to have a particularly poor prognosis. Primate animal data suggest that premenopausal CAD is strongly determined by psychosocial stress-induced central disruption of ovulatory cycling and resulting hypoestrogenemia. METHODS: We assessed reproductive hormone blood levels and angiographic CAD using core laboratories in 95 premenopausal women with coronary risk factors who were enrolled in the National Heart, Lung, and Blood Institute-sponsored Women's Ischemia Syndrome Evaluation and were undergoing coronary angiography for evaluation for suspected ischemia. RESULTS: Premenopausal women with angiographic CAD (n = 13) had significantly lower estradiol, bioavailable estradiol, and follicle-stimulating hormone (FSH) (all p < 0.05) than women without angiographic CAD (n = 82), even after controlling for age. Hypoestrogenemia of hypothalamic origin, defined as estradiol <184 pmol/l (50 pg/ml), FSH <10 IU/l, and luteinizing hormone <10 IU/l, was significantly more prevalent among the women with CAD than those without CAD (9/13 [69%] vs. 24/82 [29%], respectively, p = 0.01). Hypoestrogenemia of hypothalamic origin was the most powerful predictor of angiographic CAD in a multivariate model (odds ratio [OR] 7.4 [confidence interval (CI) 1.7 to 33.3], p = 0.008). Anxiolytic/sedative/hypnotic and antidepressant medication use were independent predictors of hypoestrogenemia of hypothalamic origin in a multivariate model (OR 4.6 [CI 1.3 to 15.7], p = 0.02, OR 0.10 [CI 0.01 to 0.92], p = 0.04, respectively). CONCLUSIONS: Among premenopausal women undergoing coronary angiography for suspected myocardial ischemia, disruption of ovulatory cycling characterized by hypoestrogenemia of hypothalamic origin appears to be associated with angiographic CAD. PMID- 12575969 TI - Effect of chelation therapy on endothelial function in patients with coronary artery disease: PATCH substudy. AB - OBJECTIVES: The purpose of this study was to evaluate the effect of chelation therapy with ethylenediamine tetraacetic acid (EDTA) on endothelium-dependent vasomotor responses in patients with documented coronary artery disease (CAD). BACKGROUND: Oxidative stress plays an important role in the dysfunction of endothelium and development of atherosclerosis. Modification of cardiac risk factors and employment of antioxidants have been shown to improve endothelial function. Ethylenediamine tetraacetic acid chelation therapy is considered to be a complementary therapy for patients with CAD and is proposed to have antioxidant properties. METHODS: A total of 47 patients enrolled in the Program to Assess Alternative Treatment Strategies to Achieve Cardiac Health (PATCH) participated in this substudy and had complete data. High-resolution ultrasound was used to assess endothelium-dependent brachial artery flow-mediated vasodilation (FMD) in patients with CAD in a randomized, double-blind, and placebo-controlled fashion. Patients were randomized to chelation therapy or placebo. The primary end point was the absolute difference in FMD after the first and 33rd treatments (6 months) of study groups compared with their baselines. RESULTS: At the baseline, the study population had mild impairment of FMD (7.2 +/- 3.4%). The first chelation treatment did not change FMD as compared with placebo (chelation 6.5 +/- 3.5% vs. placebo 7.4 +/- 2.9%; p value = 0.371). The brachial artery studies at six months did not demonstrate significant differences in FMD between study groups (placebo 7.3 +/- 3.4% vs. chelation 7.3 +/- 3.2%; p value = 0.961). CONCLUSIONS: Our results suggest that EDTA chelation therapy in combination with vitamins and minerals does not provide additional benefits on abnormal vasomotor responses in patients with CAD optimally treated with proven therapies for atherosclerotic risk factors. PMID- 12575970 TI - High-dose angiotensin-converting enzyme inhibition restores body fluid homeostasis in heart-transplant recipients. AB - OBJECTIVES: We tested the hypothesis that salt and fluid retention in heart transplant recipients (HTRs) is caused by a failure to reflexively suppress the renin-angiotensin-aldosterone system (RAAS). BACKGROUND: It is known that extracellular fluid volume is expanded (12% to 15%) in HTRs who develop hypertension. METHODS: Responses to volume expansion were measured in eight HTRs (ages 57 +/- 6 years) and six liver-transplant recipients (LTRs) (ages 52 +/- 2 years) both before and after treatment with captopril (225 mg/day). After three days of a standardized diet, 0.154 mol/l saline was infused at 8 ml/kg/h for 4 h. Blood pressure, hormones, and renal function were monitored for 48 h. After four months, the same subjects received captopril (225 mg/day), and the protocol was repeated. RESULTS: Before captopril, saline infusion suppressed the RAAS in LTRs but not in HTRs, resulting in elimination of 86 +/- 12% versus 50 +/- 11% of the sodium load by 48-h postinfusion. Blood pressure increased only in the HTRs (+16 +/- 5/9 +/- 3 mm Hg) and remained elevated for 48 h (p < or = 0.05). After captopril, sodium elimination was comparable in the liver (87 +/- 13%) and heart groups (86 +/- 12%) and blood pressure did not change in either group. CONCLUSIONS; Heart transplant recipients have blunted diuretic and natriuretic responses to volume expansion that is mediated by their inability to suppress the RAAS. Pharmacologic suppression of the RAAS normalized defects in blood pressure and fluid homeostasis. These findings indicate that hypertension in HTRs is caused, in part, by a failure to reflexively suppress the RAAS when these patients become hypervolemic. PMID- 12575971 TI - Hypertension in heart transplant recipients: more than just cyclosporine. PMID- 12575972 TI - Discrepancies between catheter and Doppler estimates of valve effective orifice area can be predicted from the pressure recovery phenomenon: practical implications with regard to quantification of aortic stenosis severity. AB - OBJECTIVES: We sought to obtain more coherent evaluations of aortic stenosis severity. BACKGROUND: The valve effective orifice area (EOA) is routinely used to assess aortic stenosis severity. However, there are often discrepancies between measurements of EOA by Doppler echocardiography (EOA(Dop)) and those by a catheter (EOA(cath)). We hypothesized that these discrepancies might be due to the influence of pressure recovery. METHODS: The relationship between EOA(cath) and EOA(Dop) was studied as follows: 1) in an in vitro model measuring the effects of different flow rates and aortic diameters on two fixed stenoses and seven bioprostheses; 2) in an animal model of supravalvular aortic stenosis (14 pigs); and 3) based on catheterization data from 37 patients studied by Schobel et al. RESULTS: Pooling of in vitro, animal, and patient data showed a good correlation (r = 0.97) between EOA(cath) (range 0.3 to 2.3 cm(2)) and EOA(Dop) (range 0.2 to 1.7 cm(2)), but EOA(cath) systematically overestimated EOA(Dop) (24 +/- 17% [mean +/- SD]). However, when the energy loss coefficient (ELCo) was calculated from EOA(Dop) and aortic cross-sectional area (A(A)) to account for pressure recovery, a similar correlation (r = 0.97) with EOA(cath) was observed, but the previously noted overestimation was no longer present. CONCLUSIONS: Discrepancies between EOA(cath) and EOA(Dop) are largely due to the pressure recovery phenomenon and can be reconciled by calculating ELCo from the echocardiogram. Thus, ELCo and EOA(cath) are equivalent indexes representing the net energy loss due to stenosis and probably are the most appropriate for quantifying aortic stenosis severity. PMID- 12575973 TI - Stenosis is in the eye of the observer: impact of pressure recovery on assessing aortic valve area. PMID- 12575974 TI - Rate-dependent effect of verapamil on atrial refractoriness. AB - OBJECTIVES: The purpose of this study was to determine whether verapamil has rate dependent effects on the atrial effective refractory period (AERP). BACKGROUND: Block of calcium current (I(Ca)) and rapid component of the delayed rectifier potassium current (I(Kr)) by verapamil is frequency-dependent. This may result in variable effects of verapamil on the AERP, depending on the rate. METHODS: The subjects of this study were 30 adults with a mean age of 45 +/- 13 years who did not have structural heart disease. In 20 subjects, the AERP was measured at basic drive cycle lengths (BDCLs) of 650 to 250 ms, in 50 ms decrements, before and after infusion of 0.1 mg/kg verapamil. The effective refractory periods (ERPs) were measured in the setting of autonomic blockade in 10 subjects and without autonomic blockade in 10 subjects. Ten subjects served as a control group and received a saline infusion instead of verapamil. RESULTS: Verapamil significantly prolonged the AERP at BDCLs of 650 to 500 ms (p < 0.01 or p < 0.05) and significantly shortened the ERP at BDCLs of 300 and 250 ms (p < 0.01). In the control group, there were no significant differences between the baseline and post-saline measurements of ERP. CONCLUSIONS: Verapamil prolongs AERP at slow rates and shortens AERP at rapid rates. These findings are consistent with a predominant effect on I(Ca) at rapid rates and a predominant effect on I(Kr) at slow rates. PMID- 12575975 TI - Duration of abnormal SPECT myocardial perfusion imaging following resolution of acute ischemia: an angioplasty model. AB - OBJECTIVES: This study was designed to determine how long nuclear myocardial perfusion imaging (MPI) remains abnormal following transient myocardial ischemia. BACKGROUND: Acute rest MPI identifies myocardial ischemia with a high sensitivity when the radionuclide is injected during chest pain. However, the sensitivity of this technique is uncertain when the radionuclide is injected following the resolution of symptoms. METHODS: Forty patients undergoing successful coronary angioplasty were randomized into four equal groups. Tc-99m sestamibi was injected intravenously during the last balloon inflation (acute MPI) in 30 patients and then reinjected 1, 2, or 3 h later (delayed MPI). In a fourth group, the radiopharmaceutical was injected at 15 min following balloon deflation (delayed MPI). A final injection was performed at 24 to 48 h (late MPI) in 37 patients (93%). RESULTS: A perfusion defect was detected in all 30 acute MPI studies; in 7/10 patients (70%) injected at 15 min; in 11/30 patients (37%) injected at 1, 2, or 3 h; and in 7/37 patients (19%) injected at 24 to 48 h. Perfusion scores were 13.0 +/- 9.2 on acute MPI, 5.1 +/- 2.8 at 15 min (p < 0.001 vs. acute MPI); 2.6 +/- 3.0 at 1, 2, and 3 h (p < 0.001 vs. acute MPI); and 1.3 +/- 2.4 at 24 to 48 h (p < 0.001 vs. acute MPI; p < 0.03 vs. delayed MPI). CONCLUSIONS: Myocardial perfusion imaging may remain abnormal for several hours following transient myocardial ischemia even when normal flow is restored in the epicardial coronary artery. PMID- 12575976 TI - Exercise training improves biventricular oxidative metabolism and left ventricular efficiency in patients with dilated cardiomyopathy. AB - OBJECTIVES: The aim of this study was to determine the effect of exercise training on myocardial oxidative metabolism and efficiency in patients with idiopathic dilated cardiomyopathy (DCM) and mild heart failure (HF). BACKGROUND: Exercise training is known to improve exercise tolerance and quality of life in patients with chronic HF. However, little is known about how exercise training may influence myocardial energetics. METHODS: Twenty clinically stable patients with DCM (New York Heart Association classes I through III) were prospectively separated into a training group (five-month training program; n = 9) and a non trained control group (n = 11). Oxidative metabolism in both the right and left ventricles (RV and LV) was measured using [(11)C]acetate and positron emission tomography. Myocardial work power was measured using echocardiography. Myocardial efficiency for forward work was calculated as myocardial work power per mass/LV oxidative metabolism. RESULTS: Significant improvements were noted in exercise capacity (VO(2)) and ejection fraction in the training group, whereas no changes were observed in the non-trained group. Exercise training reduced both RV and LV oxidative metabolism and elicited a significant increase in LV forward work efficiency, although no significant changes were observed in the non-trained group. CONCLUSIONS: Exercise training improves exercise tolerance and LV function. This is accompanied by a decrease in biventricular oxidative metabolism and enhanced forward work efficiency. Therefore, exercise training elicits an energetically favorable improvement in myocardial function and exercise tolerance in patients with DCM. PMID- 12575977 TI - Noninvasive measurements of myocardial oxygen consumption--can we do better? PMID- 12575978 TI - Red wine increases the expression of human endothelial nitric oxide synthase: a mechanism that may contribute to its beneficial cardiovascular effects. AB - OBJECTIVES: The study tested the effect of red wine on endothelial-type nitric oxide synthase (eNOS) expression and eNOS activity in human endothelial cells. BACKGROUND: Endothelial-type nitric oxide (NO) synthase exerts vasoprotective effects. Moderate alcohol consumption has been associated with a reduction of cardiovascular disease, and red wine seems to offer more benefits than any other type of drink. However, the molecular basis of this protective effect is unclear. METHODS: Human endothelial cells were treated with red wine, and eNOS messenger ribonucleic acid (mRNA) expression was measured by RNase protection assay, eNOS protein expression by Western blotting, and eNOS activity by RFL-6 reporter cell assay. The eNOS promoter activity was analyzed in transfected endothelial cells; binding activities of relevant transcription factors were determined by electrophoretic mobility shift assay. RESULTS: Incubation of endothelial cells with red wines from France upregulated eNOS mRNA and protein expression. In contrast, red wines from Germany showed little or no effect on eNOS expression. No significant difference in eNOS mRNA expression could be detected between "en barrique" (matured in oak barrels) and "non-barrique" (matured in steel tanks) produced French red wines. Endothelial cells treated with French red wines produced up to three times more bioactive NO than did control cells. French red wines increased the activity of the eNOS promoter, with the essential trans stimulated sequence being located in the proximal 326 bp of the promoter sequence. The eNOS mRNA stability was also increased by red wine. CONCLUSIONS: The increase in eNOS expression and activity brought about by red wines from France (and probably other locations) may contribute to the beneficial effects of this beverage on the cardiovascular system. PMID- 12575979 TI - Vintners and vasodilators: are French red wines more cardioprotective? PMID- 12575980 TI - Nicotine inhibits cardiac apoptosis induced by lipopolysaccharide in rats. AB - OBJECTIVES: Apoptosis develops in several heart diseases, but the therapeutic options are limited. It was hypothesized that nicotine, which inhibits apoptosis in several cells, inhibits cardiac apoptosis induced by lipopolysaccharide (LPS). BACKGROUND: Over-the-counter nicotine produces sustained levels (10 to 25 ng/ml) that may be antiapoptotic. Low levels of LPS induce apoptosis by activating tissue renin-angiotensin to stimulate angiotensin II, type 1 (AT(1)) receptors in cardiac myocytes. METHODS: Adult Sprague Dawley rats were pretreated with nicotine (6 mg/kg/day) or saline for seven to ten days (miniosmotic pumps). The LPS (1 mg/kg) was injected intravenously. Toll-like receptor 4 (TLR4) and angiotensinogen messenger ribonucleic acid (mRNA) were measured in the heart after 0, 4, 8, 16, and 24 h. Cardiac apoptosis was measured by terminal deoxy nucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining after 24 h. In vitro effects of LPS (10 ng/ml, 24 h) were studied in cardiac myocytes isolated from rats pretreated with nicotine for 7 to 10 days, or after pre exposing myocytes to nicotine (15 ng/ml) for 1, 4, 16, or 24 h. RESULTS: Neither nicotine nor LPS affected systolic blood pressure. The LPS increased cardiac apoptosis after 24 h in saline-treated, but not nicotine-treated rats, despite similar increases in cardiac TLR4 and angiotensinogen mRNA over 8 to 16 h. The LPS-induced apoptosis was blocked by pre-exposing myocytes to nicotine for 4 to 24 h (partial inhibition after 1 h). Nicotine did not inhibit apoptosis induced by angiotensin II (100 nM, 24 h). CONCLUSIONS: Therapeutic levels of nicotine inhibit LPS-induced cardiac apoptosis. This occurs after LPS increases TLR4 and angiotensinogen mRNA, but proximal to AT(1) receptor activation. Nicotine may be a novel inhibitor of cardiac apoptosis in conditions associated with circulating LPS (e.g., decompensated heart failure, acute and chronic infections). PMID- 12575981 TI - Nicotine promotes arteriogenesis. AB - OBJECTIVES: In the current study, we used a model of limb ischemia to determine whether nicotine could enhance arteriogenesis, to compare the magnitude of this effect to the angiogenic factor basic fibroblast growth factor (bFGF), and to investigate the mechanisms of the effect. BACKGROUND: We have shown previously that nicotine stimulates angiogenesis via stimulation of endothelial nicotinic cholinergic receptors. Stimulation of endothelial nicotinic cholinergic receptors causes endothelial cell proliferation, migration, and formation of capillary networks in vitro and angiogenesis in vivo in conditions of ischemia and inflammation. METHODS: New Zealand White rabbits (n = 85) underwent unilateral femoral artery occlusion and were randomized to nicotine (0.05 to 5.0 microg/kg/day), bFGF (10 microg/kg/day), or vehicle delivered intra-arterially via osmotic minipumps. At day 21, morphologic changes were assessed by immunohistochemistry and angiography. Blood flow in the ischemic limb was determined by intra-arterial Doppler flow measurements and microsphere distribution. RESULTS: Nicotine enhanced capillary density in the ischemic hind limb to a similar extent as bFGF. Nicotine also increased angiographic score, calf blood pressure ratio, intra-arterial Doppler flow, and microsphere distribution. In vitro, nicotine stimulated monocyte adhesion and transmigration. Nicotine increased by two- to three-fold the expression of monocyte adhesion molecules CD11b and CD11a; the expression of the endothelial adhesion molecule intercellular adhesion molecule-1; and the endothelial release of monocyte chemoattractant protein-1. CONCLUSIONS: In the short term, nicotine promotes angiogenesis and arteriogenesis in the setting of ischemia. The effect of nicotine may be mediated in part by activation of endothelial-monocyte interactions involved in arteriogenesis. PMID- 12575982 TI - Basic cardiovascular research and its implications for the medicinal use of nicotine. PMID- 12575983 TI - The platelet-endothelium interaction mediated by lectin-like oxidized low-density lipoprotein receptor-1 reduces the intracellular concentration of nitric oxide in endothelial cells. AB - OBJECTIVES: To address the potential role of the endothelial lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in the thrombotic system, in this study we first examined whether platelet interaction with LOX-1 generated reactive oxygen species (ROS) and superoxide (O2.-) and then investigated the relationship between the intracellular production of O2.- and the availability of nitric oxide (NO). BACKGROUND: Oxidative inactivation of NO is regarded as an important cause of its decreased biologic activity which may favor platelet dependent arterial thrombosis. METHODS: Bovine aortic endothelial cells (BAECs) and Chinese hamster ovary-K1 cells stably expressing bovine LOX-1 (BLOX-1-CHO) were incubated at different times with human platelets. The ROS, O2.-, and NO were measured in cells by flow cytometry. RESULTS: The incubation of BAECs and BLOX-1-CHO cells with human platelets induced a sharp and dose-dependent increase in intracellular concentration of ROS and O2.- (p from <0.01 to <0.001). The increase in intracellular concentration of O2.- was followed by a dose-dependent reduction in basal and bradykinin-induced intracellular NO concentration (p from <0.01 to <0.001). The increase in O2.- and the reduction of NO were inhibited by the presence of vitamin C and anti-LOX-1 monoclonal antibody (p < 0.001). CONCLUSIONS: The results of this study show that one of the pathophysiologic consequences of platelet binding to LOX-1 may be the inactivation of NO through an increased cellular production of O2.-. PMID- 12575984 TI - Atorvastatin, administered at the onset of reperfusion, and independent of lipid lowering, protects the myocardium by up-regulating a pro-survival pathway. AB - OBJECTIVES: The purpose of this study was to determine whether atorvastatin, a 3 hydroxy-3-methylglutaryl (HMG)-co-enzyme A (CoA) reductase inhibitor, limits myocardial necrosis when administered as an adjunct to reperfusion. BACKGROUND: Statins inhibit HMG-CoA reductase to reduce the synthesis of cholesterol. However, it is proposed that statins have cardiovascular effects beyond their ability to lower cholesterol, possibly via recruitment of phosphatidyl inositol 3 kinase (PI3K) and the serine/threonine kinase, Akt. This signaling pathway has recently been linked to growth factor-mediated reperfusion salvage. METHODS: Isolated perfused mouse hearts were subjected to 35 min of global ischemia and reperfused for 30 min in the presence of incremental concentrations of atorvastatin. Infarct size was determined by triphenyltetrazolium chloride staining, and the activity of the PI3K signaling cascade was determined by Western blot analysis. RESULTS: We found that there was a profound dose-dependent reduction of infarct size with atorvastatin in the range of 25 to 100 micromol/l (optimal protection was seen at 50 micromol/l with infarct size of 16 +/- 2% vs. control, 33 +/- 2%, p < 0.01). Moreover, this protection was sensitive to inhibition with the PI3 kinase inhibitor, wortmannin, and was absent in endothelial nitric oxide synthase (eNOS) knockout mice. Western blot analysis revealed that atorvastatin resulted in rapid activation of the PI3K/Akt signaling cascade (within 5 min) and that both Akt and eNOS phosphorylation were significantly increased by 4.1-fold and 2.9-fold, respectively (p < 0.01). Moreover, phosphorylation of the PI3K substrates was abrogated by the administration of wortmannin. CONCLUSIONS: Atorvastatin attenuates lethal reperfusion-induced injury in a manner that is reliant on PI3K and Akt activity and the presence and activity of eNOS. PMID- 12575985 TI - Duplicate publication: insights into the essence of a medical journal. PMID- 12575987 TI - Safety and benefits of transferring patients with acute myocardial infarction to interventional center for immediate angioplasty. PMID- 12575988 TI - Patients with high-risk acute myocardial infarction randomized to one of two treatment strategies: delay and eligibility questions. PMID- 12575989 TI - Possible risks to patients receiving statins combined with other medications. PMID- 12575990 TI - Structure of SET domain proteins: a new twist on histone methylation. AB - The methylation of lysine residues on histone tails is catalyzed by proteins containing a conserved SET domain. A recent flurry of structures of SET domain proteins has revealed a new protein fold and a scaffold for understanding catalysis and substrate binding by these enzymes. The prospect that histone methylation might form an epigenetic code and the implicated involvement of SET domain proteins in cancer assures that structure-function studies of these enzymes will continue until their detailed mechanism of action is determined. PMID- 12575991 TI - Non-homologous end-joining: bacteria join the chromosome breakdance. AB - The repair of DNA double-strand breaks by non-homologous end-joining (NHEJ) has long been thought to be restricted to eukaryotes. However, recent papers document the existence of operons encoding functional NHEJ complexes in some bacteria. These findings provide new evolutionary insights into the core biochemistry of this repair pathway, and suggest that one function driving the selection of NHEJ in bacteria, and perhaps eukaryotes, relates to prolonged periods of mitotic exit. PMID- 12575992 TI - The THAP domain: a novel protein motif with similarity to the DNA-binding domain of P element transposase. AB - We have identified a novel evolutionarily conserved protein motif - designated the THAP domain - that defines a new family of cellular factors. We have found that the THAP domain presents striking similarities with the site-specific DNA binding domain (DBD) of Drosophila P element transposase, including a similar size, N-terminal location, and conservation of the residues that define the THAP motif, such as the C2CH signature (Cys-Xaa(2-4)-Cys-Xaa(35-50)-Cys-Xaa(2)-His). Our results suggest that the THAP domain is a novel example of a DBD that is shared between cellular proteins and transposases from mobile genomic parasites. PMID- 12575993 TI - The Tudor domain 'Royal Family': Tudor, plant Agenet, Chromo, PWWP and MBT domains. AB - We have identified a family of 'Agenet' domains that are plant-specific homologs of Tudor domains. This finding has been extended, using a combination of sequence and structure-dependent approaches, to show that the three beta-stranded core regions of Tudor, PWWP, chromatin-binding (Chromo) and MBT domains are homologous because they originate from a common ancestor. In addition, we have revealed pairs of tandem repeats in the fragile X mental retardation protein (FMRP) family that are also members of this Tudor domain 'Royal Family'. PMID- 12575994 TI - The insertion of palindromic repeats in the evolution of proteins. AB - The current theory of protein evolution is that all contemporary proteins are derived from an ancestral subset. However, each new sequenced genome exhibits many genes with no detectable homologues in other species, leading to the paradoxical picture of a universal ancestor with more genes than any of its progeny. Standard explanations indicate that fast evolving genes might disappear into the 'twilight zone' of sequence similarity. Regardless of the size of the original ancestral subset, its origin and the potential mechanisms of its subsequent enlargement are rarely addressed. Sequencing of Rickettsia conorii genome recently led to the discovery of three families of repeat-mobile elements frequently inserted into the middle of protein coding genes. Although not yet identified in other species of bacteria, this discovery has provided the first clear evidence for the de novo creation of long protein segments (up to 50 amino acid residues) by repeat insertion. Based on previous results and theories on the coding potential of palindromic elements, we speculate that their insertion and mobility might have played a significant role in the early stages of protein evolution. PMID- 12575995 TI - Extended disordered proteins: targeting function with less scaffold. AB - It has been estimated that a large fraction of cellular proteins are natively disordered. Current opinion largely holds that natively disordered proteins are more 'adaptive', leading to advantages in regulation and in binding diverse ligands. Here, we argue for another, simple, physically based reason. Disordered proteins often have large intermolecular interfaces, the size of which is dictated by protein function. For proteins to be stable as monomers with extensive interfaces, protein size would need to be 2-3 times larger. This would either increase cellular crowding or enlarge the size of the cell by 15-30%, owing to the increase in the sequence length. Smaller sizes of cells, proteins, DNA and RNA conserve energy. Thus, disordered proteins provide a simple yet elegant solution to having large intermolecular interfaces, but with smaller protein, genome and cell sizes. PMID- 12575996 TI - Historical review: viruses, crystals and geodesic domes. AB - In the mid 1950s, Francis Crick and James Watson attempted to explain the structure of spherical viruses. They hypothesized that spherical viruses consist of 60 identical equivalently situated subunits. Such an arrangement has icosahedral symmetry. Subsequent biophysical and electron micrographic data suggested that many viruses had >60 subunits. Drawing inspiration from architecture, Donald Caspar and Aaron Klug discovered a solution to the problem - they proposed that spherical viruses were structured like miniature geodesic domes. PMID- 12575997 TI - Translational control by the 3'-UTR: the ends specify the means. AB - In most cases, translational control mechanisms result from the interaction of RNA-binding proteins with 5'- or 3'-untranslated regions (UTRs) of mRNA. In organisms ranging from viruses to humans, protein-mediated interactions between transcript termini result in the formation of an RNA loop. Such RNA 'circularization' is thought to increase translational efficiency and, in addition, permits regulation by novel mechanisms, particularly 3'-UTR-mediated translational control. Two general mechanisms of translational inhibition by 3' UTR-binding proteins have been proposed, one in which mRNA closure is disrupted and another in which mRNA closure is required. Experimental evidence for the latter is provided by studies of interferon-gamma-mediated translational silencing of ceruloplasmin expression in monocytic cells. A multi-species analysis has shown that, in most vertebrates, 3'-UTRs are substantially longer than their 5' counterparts, indicating a significant potential for regulation. In addition, the average length of 3'-UTR sequences has increased during evolution, suggesting that their utilization might contribute to organism complexity. PMID- 12575998 TI - Making sense of mimic in translation termination. AB - The mechanism of translation termination has long been a puzzle. Recent crystallographic evidence suggests that the eukaryotic release factor (eRF1), the bacterial release factor (RF2) and the ribosome recycling factor (RRF) all mimic a tRNA structure, whereas biochemical and genetic evidence supports the idea of a tripeptide 'anticodon' in bacterial release factors RF1 and RF2. However, the suggested structural mimicry of RF2 is not in agreement with the tripeptide 'anticodon' hypothesis and, furthermore, recently determined structures using cryo-electron microscopy show that, when bound to the ribosome, RF2 has a conformation that is distinct from the RF2 crystal structure. In addition, hydroxyl-radical probings of RRF on the ribosome are not in agreement with the simple idea that RRF mimics tRNA in the ribosome A-site. All of this evidence seriously questions the simple concept of structural mimicry between proteins and RNA and, thus, leaves only functional mimicry of protein factors of translation to be investigated. PMID- 12575999 TI - Functional domains in tetraspanin proteins. AB - Exciting new findings have emerged about the structure, function and biochemistry of tetraspanin proteins. Five distinct tetraspanin regions have now been delineated linking structural features to specific functions. Within the large extracellular loop of tetraspanins, there is a variable region that mediates specific interactions with other proteins, as well as a more highly conserved region that has been suggested to mediate homodimerization. Within the transmembrane region, the four tetraspanin transmembrane domains are probable sites of both intra- and inter-molecular interactions that are crucial during biosynthesis and assembly of the network of tetraspanin-linked membrane proteins known as the 'tetraspanin web'. In the intracellular juxtamembrane region, palmitoylation of cysteine residues also contributes to tetraspanin web assembly, and the C-terminal cytoplasmic tail region could provide specific functional links to cytoskeletal or signaling proteins. PMID- 12576000 TI - Application of a horizontal subsurface flow constructed wetland on treatment of dairy parlor wastewater. AB - Two horizontal subsurface flow reed beds of 75 m(2) each, treating dairy parlor effluent and domestic sewage (about 6.5 m(3)/day), were set-up to determine the efficiency of this system in reducing the polluting load in an isolated mountain rural settlement.A total suspended solids value of about 0.70 g/l and chemical oxygen demand (COD) and biological oxygen demand (BOD(5)) values of about 1200 and 450 mg/l O(2), respectively, were characteristic of the influent waters. Removal of suspended solids and organic load constantly remained at levels above 90%, while those of the nutrients N and P were about 50% and 60%, respectively. The total number of coliform bacteria and Escherichia coli was reduced by more than 99%, and faecal streptococci by more than 98%. Nitrates, chlorides, sulfates, anionic and non-ionic surface-active agents and heavy metals were detected only in low concentrations. Concentration and localization of metals was also quantified in Phragmites tissues by microanalysis.Results demonstrated the use of reed beds as an appropriate treatment to reduce pollutants in wastewater from rural activities to values acceptable for discharge into surface waters. PMID- 12576001 TI - Chemical composition of lipophilic extractives released during the hot water treatment of wheat straw. AB - Treatment of wheat straw with hot water at 80-95 degrees C for 0.5 h at pH 6.0 8.0 released 41.0-53.0% of the original lipophilic extractives. The chemical compositions of six lipophilic extractives were determined by GC on a medium length high-temperature capillary column without derivatization, thus giving a method for direct determination of individual components of free fatty and resin acids, sterols, waxes, sterol esters, and triglycerides. The extracts contained 68.7-75.8% lipophilic substances, comprising mainly free fatty acids (25.8 48.4%), waxes (9.4-27.0%), sterols (4.1-8.0%), triglycerides (3.3-11.0%), and sterol esters (2.6-5.1%). Minor amounts of diglycerides (0.3-0.5%), resin acid (0.5-3.1%), and phenolic compounds (0.9-3.6%) were also quantitatively determined in the extractives. PMID- 12576002 TI - Optimization of lipase-catalyzed biodiesel by response surface methodology. AB - Biodiesel prepared by catalyzed mild transesterification has become of much current interest for bioenergy. The ability of a commercial immobilized lipase (Novo Industries--Bagsvaerd, Denmark) from Rhizomucor miehei (Lipozyme IM-77) to catalyze the transesterification of soybean oil and methanol was investigated in this study. Response surface methodology and 5-level-5-factor central composite rotatable design were employed to evaluate the effects on reaction time, temperature, enzyme amount, molar ratio of methanol to soybean oil, and added water content on percentage weight conversion to soybean oil methyl ester by transesterification. Based on ridge max analysis, the optimum synthesis conditions giving 92.2% weight conversion were: reaction time 6.3 h, temperature 36.5 degrees C, enzyme amount 0.9 BAUN (Batch Acidolysis Units NOVO), substrate molar ratio 3.4:1, and added water 5.8%. PMID- 12576003 TI - Nitrogen removal from wastewater using a double-biofilm reactor with a continuous flow method. AB - Wastewater microorganisms of nitrification and denitrification were cultivated to compose two biofilm modules, termed the permeable support bioreactor (PSB) and the membrane feeding substrate bioreactor (MFSB). PSB and MFSB were combined in a single tank to develop a double-biofilm reactor, which was used to treat nitrogen contaminants in wastewater. With a membrane supplement of substrates (O(2) and CH(3)OH), the D.O. and COD levels were at a low value in the bulk solution thus inhibitive effects between nitrification and denitrification were minimized. Simultaneous nitrification/denitrification was conducted in the reactor and the double-biofilm reactor achieved high nitrification and denitrification efficiency, of 96.5% and 82%, respectively. PMID- 12576004 TI - Decolourization of azo dyes and a dye industry effluent by a white rot fungus Thelephora sp. AB - A white rot fungus Thelephora sp. was used for decolourization of azo dyes such as orange G (50 microM), congo red (50 microM), and amido black 10B (25 microM). Decolourization using the fungus was 33.3%, 97.1% and 98.8% for orange G, congo red and amido black 10B, respectively. An enzymatic dye decolourization study showed that a maximum of 19% orange G was removed by laccase at 15 U/ml whereas lignin peroxidase (LiP) and manganese dependent peroxidase (MnP) at the same concentration decolourized 13.5% and 10.8%, orange G, respectively. A maximum decolourization of 12.0% and 15.0% for congo red and amido black 10B, respectively, was recorded by laccase. A dye industry effluent was treated by the fungus in batch and continuous modes. A maximum decolourization of 61% was achieved on the third day in the batch mode and a maximum decolourization of 50% was obtained by the seventh day in the continuous mode. These results suggest that the batch mode of treatment using Thelephora sp. may be more effective than the continuous mode for colour removal from dye industry effluents. PMID- 12576005 TI - Energy evaluation of forest residues originated from pine in Galicia. AB - Pine residues originated from cuts and forestry tasks were studied with the main objective of using them as an alternative source of energy. Calorific values were measured using a static bomb calorimeter in an oxygen atmosphere. From these measurements a value of 7500 kJkg(-1) for the mean lower heating value of these kind of residues was found. A rational and sustainable exploitation of the calorific energy contained in these residues can yield a gross benefit of 2.8 x 10(8) per year through its transformation in electric energy. The study was made along one year, using samples collected in different previously characterized zones. Some other parameters such as: elementary composition, heavy metal content, flammability, and bioclimatic properties were also determined. PMID- 12576006 TI - Performance of a glucose fed periodic anaerobic baffled reactor under increasing organic loading conditions: 1. Experimental results. AB - The influence of the organic loading rate on the performance of an innovative reactor, the periodic anaerobic baffled reactor (PABR) was examined. A laboratory scale PABR of four compartments being fed with a glucose based synthetic medium performed with high stability while the feed organic load was doubled from 12.5 to 25 and then to 50 gCOD/l. Finally the feed concentration was increased to 75 gCOD/l. The successive step changes in the feed concentration lasted for 20, 15, and 7 d, respectively. The COD removal efficiency of the PABR was satisfactory in the first two transitions (approximately 97.5 and 96%). In the third transition (OLR=18.75 gCOD/l/d) the reactor failed as the pH dropped to 4. The concentrations of butyric and valeric acids increased as the organic loading was increased and eventually they became greater than the concentration of acetic and propionic acids. PMID- 12576007 TI - Performance of a glucose fed periodic anaerobic baffled reactor under increasing organic loading conditions: 2. Model prediction. AB - A model was developed for the anaerobic digestion of a glucose-based medium in an innovative high-rate reactor, the periodic anaerobic baffled reactor (PABR). The model considers each PABR compartment as two variable volume interacting sections, of constant total volume, one with high solids and one with low solids concentration, with the gas and liquid flows influencing the material flows between the two sections. For the simulation of glucose degradation, the biomass was divided into acidogenic, acetogenic and methanogenic groups of microorganisms. The kinetic part of the model accounted for possible inhibition of acidogenesis, acetogenesis and methanogenesis by volatile fatty acids. The model succeeded in predicting the reactor performance upon step increases in the organic loading rate. PMID- 12576008 TI - Comparison of optimised isotherm models for basic dye adsorption by kudzu. AB - This study assesses the use of dried (5% w/w moisture) kudzu (Peuraria lobata ohwi) as an adsorbent medium for the removal of two basic dyes, Basic Yellow 21 and Basic Red 22, from aqueous solutions. The extent of adsorption was measured through equilibrium sorption isotherms for the single component systems. Equilibrium was achieved after 21 days. The experimental isotherm data were analysed using Langmuir, Freundlich, Redlich-Peterson, Temkin and Toth isotherm equations. A detailed error analysis was undertaken to investigate the effect of using different error criteria for the determination of the single component isotherm parameters. The performance of the kudzu was compared with an activated carbon (Chemviron F-400). Kudzu was found to be an effective adsorbent for basic dye colour removal; though its capacity for colour removal was not as high as an activated carbon, the potential appeared to exist to use it as an alternative to activated carbon where carbon cost was prohibitive. PMID- 12576009 TI - Using tea waste as a new casing material in mushroom (Agaricus bisporus (L.) Sing.) cultivation. AB - In this study, the possibility of using tea production waste as a new casing material in mushroom (Agaricus bisporus) cultivation was investigated. Some physical and chemical characteristics of tea waste, fermented tea waste and a mixture of tea waste with peat were compared with that of peat casing, as were their effects on yield. The highest yield was obtained from peat casing. Using tea production waste alone as a casing was not acceptable for assured yield when it was compared with peat. But, a mixture of tea production waste with peat in 1:1 (v:v) ratio increased the yield. There was no significant difference between the mushroom yields of tea production waste+peat and peat casing materials at the end of 30 and 40 days. High salt content, organic and inorganic compounds in casing materials caused reduction of yields. However, a high iron content in casing material gave a significant positive correlation with total yield at 40 days. PMID- 12576010 TI - Potential of biogas recirculation to enhance biomass accumulation on supporting media. AB - Two lab-scale anaerobic hybrid reactors (AHR) were operated to investigate the effect of recirculated biogas on the development of biomass on supporting media during the start-up. The reactor comprised of two distinct zones; sludge bed on the bottom and packed bed using nylon fiber as the media on the upper half of the reactor. Both reactors were continuously fed with cassava starch wastewater. The organic loading rate (OLR) was increased from 0.3 to 5.5 g COD/L/day by gradually decreasing the hydraulic retention time (HRT) from 37 to 3.5 days in two months. The biogas at 2.6 L/L/day was recirculated merely in the first month of the operation in order to allow the attached biomass to grow according to the organic matters present in the reactor at the final stage of the start up. Chemical oxygen demand (COD) removal efficiency of over 80% was achieved throughout the study. The result demonstrated a better COD removal efficiency for the reactor with biogas recirculation, especially at low HRTs. The amounts of biomass accumulated on the media in both reactors were slightly different with 11.9 gVSS found on the one with biogas recirculation compared to 9.8 gVSS on the other. In addition, 16.3% increase of the sludge bed was achieved with biogas recirculation as opposed to 9% in the control one. The attached biomass activity test indicated a greater amount and more favorable ratio of the methanogenic bacterial group on the media with the recirculation correlating well to a relatively higher methane content in biogas. As a result, the recirculation of biogas has a potential of improving the characteristics of the AHR especially in terms of biomass accumulation. PMID- 12576011 TI - Chlorella sorokiniana immobilized on the biomatrix of vegetable sponge of Luffa cylindrica: a new system to remove cadmium from contaminated aqueous medium. AB - A new sorption system of microalgal cells immobilized on the biostructural matrix of Luffa cylindrica for sequestering cadmium is reported. Free and immobilized Chlorella sorokiniana removed cadmium from 10 mgl(-1) solution at the efficiency of 92.7% and 97.9% respectively. Maximum cadmium sorption was observed to be 39.2 mgg(-1) at equilibrium (C(eq)) of 112.8 mgl(-1) by immobilized microalgal biomass as compared to 33.5 mgg(-1) at C(eq) of 116.5 mgl(-1) by free biomass from initial concentration of 150 mgl(-1). In continuous liquid flow column, the cadmium sorption capacity of immobilized C. sorokiniana was 192 mgg(-1), which was 73.2% of the total metal passed in 51.5 l. Metal desorption with 0.1 M HCl was 100% and the desorbed immobilized system was reusable with a similar efficiency in the subsequent cycle. PMID- 12576013 TI - Paper of the Year Award. PMID- 12576015 TI - A second response in correcting the HslV-HslU quaternary structure. PMID- 12576016 TI - X-ray microCT study of pyramids of the sea urchin Lytechinus variegatus. AB - This paper reports results of a novel approach, X-ray microCT, for quantifying stereom structures applied to ossicles of the sea urchin Lytechinus variegatus. MicroCT, a high resolution variant of medical CT (computed tomography), allows noninvasive mapping of microstructure in 3-D with spatial resolution approaching that of optical microscopy. An intact pyramid (two demipyramids, tooth epiphyses, and one tooth) was reconstructed with 17 microm isotropic voxels (volume elements); two individual demipyramids and a pair of epiphyses were studied with 9-13 microm isotropic voxels. The cross-sectional maps of a linear attenuation coefficient produced by the reconstruction algorithm showed that the structure of the ossicles was quite heterogeneous on the scale of tens to hundreds of micrometers. Variations in magnesium content and in minor elemental constitutents could not account for the observed heterogeneities. Spatial resolution was insufficient to resolve the individual elements of the stereom, but the observed values of the linear attenuation coefficient (for the 26 keV effective X-ray energy, a maximum of 7.4 cm(-1) and a minimum of approximately 2 cm(-1) away from obvious voids) could be interpreted in terms of fractions of voxels occupied by mineral (high magnesium calcite). The average volume fraction of mineral determined for a transverse slice of the demipyramid near where it joins an epiphysis was 0.46; for a slice 3.3 mm adoral it was 0.70. Local volume fractions of mineral approached 1, and, away from resolvable voids, considerable portions of the demipyramids had volume fractions of calcite at or below approximately 0.33. MicroCT imaging of a demipyramid before and after infiltration with a high absorptivity fluid (sodium polytungstate) confirmed the determination of the volume fractions of minerals. PMID- 12576017 TI - Structural differences between "dark" and "bright" isolated human osteonic lamellae. AB - This investigation presents new insights into the structure of human secondary lamellae. Lamellar specimens that appear dark and bright on alternate osteon transverse sections under circularly polarizing light were isolated using a new technique, and examined by polarizing light microscopy, synchrotron X-ray diffraction, and confocal microscopy. A distribution of unidirectional collagen bundles and of two overlapping oblique bundles appears on circularly polarizing light microscopy images in relation to the angle between the specimen and the crossed Nicols' planes. The unidirectional collagen bundles observed at 45 degrees run parallel to the osteon axis in the dark lamellar specimens and perpendicular to it in the bright ones. Small and wide-angle micro-focus X-ray diffraction indicates that the dark lamellae are structurally quite homogeneous, with collagen fibers and apatite crystals preferentially oriented parallel to the osteon axis. Bright lamellar specimens exhibit different orientation patterns with the dominant ones bidirectional at +/-45 degrees with respect to the osteon axis. Accordingly, confocal microscopy evidences the presence of longitudinal bundles in dark lamellar specimens and oblique bundles in the bright ones. Radial bundles are evidenced in both lamellar types. The alternate osteon structure is described by a rather continuous multidirectional pattern, in which dark and bright lamellae display different mechanical and possibly biological functions. PMID- 12576018 TI - Pyrodictium cannulae enter the periplasmic space but do not enter the cytoplasm, as revealed by cryo-electron tomography. AB - The hyperthermophilic archaeon Pyrodictium grows in the form of a macroscopically visible network. It consists of cells entrapped in an extracellular matrix of hollow tubules, the "cannulae." Here, we present the three-dimensional structure of a single cell in conjunction with two extracellular cannulae, as determined by cryo-electron microscopy. To achieve this, the information from two independent tilt series of the same specimen was combined, with the specimen rotated in the second series. In the three-dimensional tomographic reconstruction, we were able to trace the two cannulae in their full length, in particular, also inside the cell. One cannula enters the periplasmic space, while the other cannula contacts the surface of the cell, the S-layer. This indicates that the cannulae interconnect individual cells with each other on the level of their periplasmic space; we do not, however, have evidence that they enter the cytoplasm of the cells. The implications of these data for possible functions of the cannulae are discussed. PMID- 12576019 TI - Capturing time-resolved changes in molecular structure by negative staining. AB - Imaging structural intermediates of biological processes is a key step in understanding biological function. Because intermediates are commonly short lived, lasting only milliseconds, the main methods used to capture them have been conventional imaging of analog or inhibited states, having extended lifetimes, or rapid (millisecond timescale) freezing of intermediates with subsequent observation by cryo-EM. We have developed a simpler method that fixes structure on the millisecond timescale. The procedure consists of briefly (milliseconds) exposing the macromolecular structure of interest on an EM grid to conditions that initiate the structural change, then immediately fixing with uranyl acetate or tannic acid. Specimens are then observed by negative staining. The key finding that validates this approach is our demonstration that uranyl acetate, and in some cases tannic acid, fixes protein molecular structure on the millisecond timescale. This is demonstrated by our observation that exposure of actin and myosin filaments to these fixatives for as little as 10 ms is sufficient to fully preserve them against changes that normally induce rapid and major alteration in their molecular structure. Fixation appears to stabilize both ionic and hydrophobic bonds. This approach should be of general utility for studying transient molecular changes in many systems. PMID- 12576020 TI - 3-dimensional imaging of collagen using second harmonic generation. AB - Collagen is the most important structural protein of the animal body. Its unique triple-helix structure and extremely high level of crystallinity make it exceptionally efficient in generating the second harmonic of incident light, and we show here how this leads to a novel mode of microscopy of immediate practical significance in medicine and biology. In particular, it provides sensitive and high-resolution information on collagen distribution, discriminates between type I and type III collagen, and allows both a greater understanding of and a sensitive test for cirrhosis of the liver. Future research applications could include wound healing and hereditary collagen diseases such as osteogenesis imperfecta. PMID- 12576021 TI - A core-weighted fitting method for docking atomic structures into low-resolution maps: application to cryo-electron microscopy. AB - Cryo-electron microscopy of "single particles" is a powerful method to analyze structures of large macromolecular assemblies that are not amenable to investigation by traditional X-ray crystallographic methods. A key step in these studies is to obtain atomic interpretations of multiprotein complexes by fitting atomic structures of individual components into maps obtained from electron microscopic data. Here, we report the use of a "core-weighting" method, combined with a grid-threading Monte Carlo (GTMC) approach for this purpose. The "core" of an individual structure is defined to represent the part where the density distribution is least likely to be altered by other components that comprise the macromolecular assembly of interest. The performance of the method has been evaluated by its ability to determine the correct fit of (i) the alpha-chain of the T-cell receptor variable domain into a simulated map of the alphabeta complex at resolutions between 5 and 40 A, and (ii) the E2 catalytic domain of the pyruvate dehydrogenase into an experimentally determined map, at 14 A resolution, of the icosahedral complex formed by 60 copies of this enzyme. Using the X-ray structures of the two test cases as references, we demonstrate that, in contrast to more traditional methods, the combination of the core-weighting method and the grid-threading Monte Carlo approach can identify the correct fit reliably and rapidly from the low-resolution maps that are typical of structures determined with the use of single-particle electron microscopy. PMID- 12576022 TI - Bioinformatic analysis of ClpS, a protein module involved in prokaryotic and eukaryotic protein degradation. AB - ClpS is a small protein, usually encoded immediately upstream of ClpA in the genomes of proteobacteria. Recent results show that it is a molecular adaptor for substrate recognition by ClpA in Escherichia coli. We analyzed ClpS by bioinformatic methods and found that ClpS homologs are also found in organisms that lack ClpA, such as actinobacteria, cyanobacteria, and plant chloroplasts. Furthermore, ClpS is homologous to a domain in the eukaryotic E3 ubiquitin ligase, N-recognin. This domain has previously been described as responsible for the recognition of type 2 N-end rule substrates. Despite very low levels of sequence similarity to proteins of known structure, there appears to be substantial structural similarity between ClpS and the C-terminal domain of ribosomal protein L7/12 (1CTF). PMID- 12576023 TI - The variance of icosahedral virus models is a key indicator in the structure determination: a model-free reconstruction of viruses, suitable for refractory particles. AB - A model-free method to determine the three-dimensional structure of icosahedral viruses is described. The novel strategy is based upon the approximate principle that correct virus structures have high variance as do all other well-detailed structures, even wrong ones. The original projections of individual particles are reduced to a radius of 25 pixels and are used to compute single particle reconstruction models by assigning them 1800 different Euler triads. The variance of the models obtained from all projections is stored in maps and a decimation process is carried out. In a first stage, thresholds are adopted for the variance values, and in a second stage, carried out by correspondence analysis and classification, 30 clusters of models are sorted out. The clusters are refined to yield models contained in boxes of 64(3) voxels. The refined models with highest variance and closest similarity represent the correct solution. Once enlarged, these models can be used to align all available projections in their original scale in a customary projection-matching process. The method has proved successful in determining the structures of poliovirus, of the empty and filled capsids of L-A virus, and of a modified capsid of hepatitis B virus. PMID- 12576024 TI - Crystallization and preliminary X-ray diffraction analysis of monoprenylated Rab7 GTPase in complex with Rab escort protein 1. AB - Posttranslational geranylgeranylation of Rab GTPases is catalyzed by Rab geranylgeranyltransferase (RabGGTase), which consists of a catalytic alpha/beta heterodimer and an accessory Rab escort protein (REP). REP functions as a molecular chaperone that presents Rab proteins to the RabGGTase and after prenylation delivers them to their target membrane. Mutations in the REP-1 gene in humans lead to an X-chromosome-linked defect known as choroideremia, a progressive disease that inevitably culminates in complete blindness. Here we report in vitro assembly, purification, and crystallization of the monoprenylated Rab7GDP:REP-1 complex. X-Ray diffraction data for the REP-1:Rab7 complex were collected to 2.2-A resolution at the ESRF. The crystals belong to the orthorhombic space group P2(1)2(1)2 with unit-cell parameters a=64.3A, b=105.3A, c=132.6A. Preliminary structural analysis revealed the presence of one complex in the asymmetric unit. To understand the conformational changes in Rab protein on complex formation we also crystallized the GDP-bound form of Rab7 that diffracted to at least 1.8A on the in-house X-ray source. PMID- 12576025 TI - Differential secretion of cytokines and adhesion molecules by HUVEC stimulated with low concentrations of bleomycin. AB - Bleomycin (BLM) is known to induce lung inflammation and subsequent fibrosis. Endothelial cells have been reported to play an important role, producing cytokines and adhesion molecules during the inflammatory process in pulmonary fibrosis. To examine the effects of BLM on endothelial cells, we investigated the expression profiles of various cytokines and adhesion molecules produced by endothelial cells stimulated with BLM. Increased expressions of interleukin-8 and monocyte chemoattractant protein-1 measured as protein as well as mRNA by human umbilical vein endothelial cells (HUVECs) were detected after exposure to BLM. Similarly, increased expressions of E-selectin and intercellular adhesion molecule-3 were detected both at the protein and mRNA levels. Under these conditions, a small but significant decrease of [3H]thymidine uptake was detected. These findings indicate that HUVEC were stimulated to secrete cytokines and express adhesion molecules in the presence of low concentrations of BLM which have a mildly inhibitory effect on cellular proliferation. PMID- 12576026 TI - Suppression of antibody production by TNF-related apoptosis-inducing ligand (TRAIL). AB - TNF-related apoptosis-inducing ligand (TRAIL), a type II membrane protein belonging to the TNF family, induces apoptotic cell death in various types of tumor cells. However, little is known about its pathological and physiological functions in the immune system. In this study, we showed that administration of neutralizing anti-TRAIL mAb markedly increased serum auto-Ab levels, particularly of IgG1 subclass, in autoimmune-prone C3H/HeJ gld/gld mice without affecting lymphocytosis and lymphocytes populations. In an experimental system where TNP specific Ab production was induced by immunization with TNP-modified syngeneic B lymphoma cells, expression of TRAIL on these cells significantly reduced TNP specific Ab production, especially of IgG1 subclass, without affecting T cell priming. These results suggest a new role for TRAIL in the suppression of Ab production. PMID- 12576027 TI - Attenuation of airway hyperresponsiveness in a murine asthma model by neutralization of granulocyte-macrophage colony-stimulating factor (GM-CSF). AB - Asthma is recognized as an inflammatory disease in which various cytokines are involved. Among these, granulocyte-macrophage colony-stimulating factor (GM-CSF) is known to play a critical role in the survival of eosinophils and in the activation of antigen-presenting cells (APC). We studied the effects of neutralization of GM-CSF in a murine model of asthma, to elucidate its role in enhanced airway responsiveness and in airway inflammation. A/J mice, which are genetically predisposed to acetylcholine hyperresponsiveness, were immunized with ovalbumin (OA) and alum. Thereafter, the mice were subjected to a two-week regimen of OA inhalation, during which either goat anti-mouse polyclonal GM-CSF antibody or isotype control goat IgG was administered intranasally. Pulmonary function was then analyzed using whole body plethysmography before and after acetylcholine (Ach) inhalation. Here we show that OA inhalation following OA immunization increased airway responsiveness to acetylcholine and induced GM-CSF as well as IL-4 and IL-5 mRNA expression in the lung. The administration of GM CSF-neutralizing antibody during OA inhalation significantly reduced this increased airway hyperresponsiveness and also inhibited airway inflammation. Thus, endogenous GM-CSF plays an important role in the process of airway inflammation and airway hyperresponsiveness after antigen-specific immunity has been established. PMID- 12576028 TI - Activation-induced cell death in human T cells is a suicidal process regulated by cell density but superantigen induces T cell fratricide. AB - Repeated ligation of the TCR results in apoptosis (activation-induced cell death; AICD). Superantigens such as Staphylococcal enterotoxin B (SEB) are particularly efficient at inducing AICD in T cells. We investigated whether apoptosis in human T cell subsets was due to fratricide (killing of neighboring cells) or suicide (cell autonomous death). AICD of Th1, Th2, Tc1, and Tc2 effector cells was dramatically enhanced at low cell densities and could be observed in single cell microcultures. AICD was unaffected by adhesion molecules or neighboring cells undergoing AICD, confirming the predominance of a suicidal mechanism. However, SEB was able to induce fratricidal apoptosis of type 1, but not type 2 cells. Fratricide was also observed when unstimulated T cells were exposed to activated Tc1 effector cells. Thus, AICD is tightly regulated to allow clonal T cell expansion and memory cell generation, but superantigens may subvert this process by allowing T cell fratricide. PMID- 12576029 TI - Herpetic stromal keratitis in the absence of viral antigen recognition. AB - Herpetic stromal keratitis (HSK), resulting from ocular infection with herpes simplex virus (HSV), is thought to represent a T cell mediated immunopathologic lesion. Antigens recognized by the inflammatory T cells remain unresolved and non TCR mediated activation of T cells (bystander activation) is considered as also involved. This report documents further evidence for the bystander activation mechanisms using three T cell transgenic RAG-/- mouse strains. Accordingly HSK occurred in PCC RAG-/-, P14 RAG-/-, and OT-1 RAG-/- mice. In none of the models could HSV specific T cell reactivity be demonstrated and animals were unprotected from lesion development by immunization prior to HSV ocular infection. The results support the role of bystander activation as a mechanism of T cell mediated immunopathology and show that CD8(+) as well as CD4(+) T cells can participate in HSK lesion development. PMID- 12576030 TI - Role of cell cycle regulator p19ARF in regulating T cell responses. AB - Although it is well established that the processes of cellular proliferation and apoptosis are linked, the role of cell cycle regulators in T cell responses in vivo is not well understood. In recent years, tumor suppressor molecule p19(ARF) has emerged as a key cell cycle regulator important in cellular apoptosis against strong mitogenic stimuli. In this study, we compared the antigen-specific T cell responses between wild type (+/+) and p19(ARF)-deficient (p19-/-) mice following an acute infection with lymphocytic choriomeningitis virus (LCMV). p19-/- mice mounted a potent CD8 T cell response and the magnitude of expansion of LCMV specific CD8 T cells was comparable to that of +/+ mice. Further, the clonal downsizing of the expanded virus-specific CD8 T cells and establishment of long term T cell memory were minimally affected by p19(ARF) deficiency. Therefore, p19(ARF) function is not essential to regulate T cell responses following an acute viral infection. PMID- 12576031 TI - Interaction of monocytes with vascular endothelial cells synergistically induces interferon gamma-inducible protein 10 expression through activation of specific cell surface molecules and cytokines. AB - To further understand the regulatory mechanisms involved in the process of angiogenesis, the present study was designed to determine the expression and regulation of interferon gamma-inducible protein 10 (IP-10) in peripheral blood monocytes and human umbilical vein endothelial cells (HUVECs). We found that the interaction of monocytes with HUVECs resulted in synergistic increases in IP-10 expression and secretion, which consequently inhibited endothelial tube formation in vitro. Induction of IP-10 was mediated via specific cell surface molecules, as indicated by the finding that IP-10 secretion was significantly inhibited by anti CD40 ligand antibody, and to a lesser extent by anti-CD40 antibody. Furthermore, we examined the effects of soluble mediators, such as inflammatory and immune cytokines on IP-10 secretion. Addition of interleukin (IL)-1, as well as interferon gamma, induced a marked augmentation of IP-10 secretion by unstimulated monocytes, unstimulated HUVECs, and co-cultures of the two cell types. In contrast, IL-10, recognized as an anti-inflammatory cytokine, significantly inhibited IP-10 secretion by co-cultures. Our results suggest that the interaction of monocytes with endothelial cells results in synergistic increases in IP-10 expression and secretion, which contribute to the regulation of angiogenesis and initiation of inflammatory vascular diseases. PMID- 12576032 TI - Oral metastases: report of 24 cases. AB - AIM: To study patients with oral metastatic tumours for the distribution of sex and age, the oral site and histopathological type of the metastasis, the primary tumour site and length of follow-up. PATIENTS AND METHODS: All patients who had an oral metastasis diagnosed during the period January 1970-January 2001 at the Department of Oral and Maxillofacial Surgery/Oral Pathology, VU University Medical Centre, Amsterdam, The Netherlands, were included in this retrospective case study. RESULTS: Of 1537 patients with newly diagnosed oral cancers, 24 had metastatic tumours. There was an equal sex distribution and age at the time of diagnosis ranged from 8 to 90 years (median 60). The metastatic tumours most commonly involved the bone (18/24), the mandible being the most common (15/18). The predominant histological type was adenocarcinoma. In most patients (n = 16) the primary tumour was already known before the oral metastatic lesion appeared. The most common primary tumours were breast, lung, kidney, and prostate, in that order. Prognosis was poor (median survival 6 months, range 1-60). CONCLUSIONS: Oral metastases are rare and may present at any age in both sexes and predominantly involve bony structures, particularly the mandible. A third of oral metastases appeared to be the first indication of an occult malignant process elsewhere. PMID- 12576033 TI - Shoulder complaints after neck dissection; is the spinal accessory nerve involved? AB - The purpose of the current study was to investigate the relation between shoulder morbidity (pain and range of motion), and the function of the spinal accessory nerve after neck dissection. Identifying dysfunction of the nerve gives insight in the mechanisms of post-operative shoulder complaints. In total 112 patients after neck dissection (73 males/39 females), mean (SD) age 61 (13) years, participated in the study. The mean duration of follow up was 3 (2) years. Five patients had radical, 43 modified radical, 48 supraomohyoid, and 16 posterolateral neck dissection. Thirty-nine complained of shoulder pain of whom 20 (51%) had dysfunction of the spinal accessory nerve, and 19 (49%) did not. In total 29 patients (26%) had dysfunction of the spinal accessory nerve of whom 20 (69%) had shoulder pain. Shoulder pain was significantly related to dysfunction of the nerve (P < 0.001). Twenty-three patients had a difference in active range of motion in shoulder abduction of > or =40 degrees, of whom 22 (96%) had dysfunction of the nerve. A difference in active shoulder abduction of > or =40 degrees was significantly related to loss of function of the spinal accessory nerve (P < 0.001). CONCLUSION: Shoulder pain after neck dissection can only be attributed to dysfunction of the spinal accessory nerve in about 50%. If patients experience shoulder pain after neck dissection examination of the trapezius muscle and active bilateral abduction of the shoulder should be made to find out if the spinal accessory nerve is involved. PMID- 12576034 TI - Stability of the sternoclavicular joint. A retrospective study. AB - Enlargement of the sternoclavicular joint is a well-documented but little recognised complication of radical neck dissection [AJR 3 (1971) 584]. We examined the stability of the sternoclavicular joint in 61 patients who had had radical neck dissection, functional neck dissection or sternomastoid division in the treatment of torticollis. Our findings support the hypothesis that postoperative stability of the sternoclavicular joint depends on the integrity of the accessory nerve and probably the proprioceptive branches of C3 and C4 of the cervical plexus. We conclude that in patients who require surgical section of the sternomastoid muscle in the treatment of torticollis, or for venous access in microvascular reconstruction, enlargement of the sternoclavicular joint should not occur as a postoperative complication, unlike those patients who have radical neck dissection with resection of the accessory nerve. PMID- 12576035 TI - Factors that influence the outcome of salvage in free tissue transfer. AB - INTRODUCTION: The success of salvage techniques for free tissue transfer is well documented. The aim of this study was to identify factors that influenced the results of salvage operations in a group of patients who required early exploration. METHODS: From a database survey of 408 patients who had a total of 427 free tissue transfer reconstructions, 65 (16%) returned to the operating theatre within 7 days. A retrospective analysis of their progress was made from the case records. RESULTS: The flap chart was found to be highly accurate for the 65 patients who had returned to the operating theatre, reporting two false positives and one false negative. Forty patients had compromised flaps and 25 had haematomas that required evacuation. The commonest problem with flaps was venous congestion (33/40, 83%), and 29 flaps were successfully salvaged (73%). Most successful salvage attempts were made within 24 hours of the end of the initial operation. The salvage rate was higher for the radial fasciocutaneous flaps (25/30, 83%), than for composite flaps (2/7, 29%). A total of 24/427 flaps failed (6%). CONCLUSIONS: We now recommend hourly observations of the flap for 24 hours followed by 4-hourly monitoring for 48 hours. Improved monitoring techniques for composite grafts may result in more being salvaged and a better overall survival. PMID- 12576036 TI - Post-traumatic orbital cellulitis. AB - Orbital cellulitis is uncommon. It may arise as a sequel to eyelid infection, or from direct spread of infection from the paranasal sinuses; it may be of odontogenic origin and has been reported after meningitis and after nasoorbital fractures with pre-existing sinusitis. Clinically, orbital cellulitis is of great importance, as it is a severe disease with potentially disastrous consequences. It may lead to optic neuritis, optic atrophy, blindness, cavernous sinus thrombosis, superior orbital fissure syndrome, meningitis, subdural empyema, and even death. We report two cases of severe post-traumatic orbital cellulitis with subperiosteal abscesses. These were managed surgically and vision was preserved. We describe the anatomy, a classification of orbital infection, and the importance of multidisciplinary management of these cases. PMID- 12576037 TI - An audit of 405 temporomandibular joint arthrocentesis with intra-articular morphine infusion. AB - The management of refractory pain in the temporomandibular joint (TMJ) is both challenging and controversial. Arthrocentesis is a simple technique that can be used instead of more invasive procedures in patients with pain that fails to respond to conventional conservative measures. We undertook an audit of 405 arthrocenteses in 298 patients over a 10-year period who had refractory pain in the TMJ. The pain was assessed subjectively by a visual analogue scale, both before arthrocentesis and at 1 and 6 months, and 1 year afterwards. A significant reduction in pain score was found after arthrocentesis (P < 0.001) and 269 patients (90%) found the procedure beneficial. We recommend arthrocentesis as an effective, minimally invasive technique in patients with continuing pain in the TMJ that is unresponsive to conservative management. PMID- 12576038 TI - A cost-utility analysis of patients undergoing orthognathic treatment for the management of dentofacial disharmony. AB - INTRODUCTION: Management of dentofacial discrepancies using orthognathic treatment is now a common procedure in the United Kingdom. Although the benefits of orthognathic intervention are often considered, the cost implications have not been investigated to our knowledge. This study is a cost-utility analysis of orthognathic treatment. PATIENTS AND METHODS: Twenty-one patients were interviewed five times during treatment using the time trade-off (TTO) method to establish utility values. Quality adjusted life years (QALYs) gained as a result of treatment were calculated and discounted. The resource use was calculated for each of the 21 patients individually and the costs subjected to both a sensitivity analysis and discounting. The incremental mean cost per additional QALY was calculated (as compared with a 'no treatment' approach). RESULTS: The incremental cost for each additional QALY was 561 pounds sterling for the groups combined, based on mean additional costs and QALYs (546 pounds sterling for the bimaxillary group and 617 pounds sterling for the single jaw group). DISCUSSION: Orthognathic treatment seems to provide good outcomes at relatively low cost. Even allowing for the uncertainty in mean costs and QALYs, there is a high probability of treatment being cost-effective. Cost-utility analysis is still a relatively new technique in dentistry and further studies should be encouraged. PMID- 12576039 TI - Quantitative thermosensory testing of the lingual and inferior alveolar nerves in health and after iatrogenic injury. AB - Quantitative testing of the sensory thermal thresholds testing was applied at sites innervated by the third division of the trigeminal nerve in 20 patients with no reported sensory impairment and in 20 subjects with iatrogenic nerve injuries after third molar surgery. In the control group sensitivity to cooling was significantly greater than the sensitivity to warming at all sites. The labial mucosa innervated by the inferior alveolar nerve was significantly more sensitive to thermal changes than either the mental region or the lingual mucosa. At sites supplied by nerves that had been injured, there were raised thresholds to both warming and cooling compared with the control group, and with uninjured contralateral sites. The results indicate that this test can identify iatrogenic lingual and inferior alveolar nerve injury with reference to a control group but because of spatial variation selection of control sites for comparison should be done cautiously. PMID- 12576040 TI - Patient-centred outcome measures in oral surgery: validity and sensitivity. AB - The performances of patient-centred outcome measures after oral surgery were evaluated in a prospective cohort study of 100 patients who had third molar extractions. Participants self-completed a questionnaire incorporating a general health (12-item short form health survey: SF-12) and two specific outcome scales to oral health (oral health impact profile: OHIP-14, and The United Kingdom Oral Health related Quality of Life measure: OHQoL-UK) preoperatively, daily during the immediate postoperative period, and at the review appointment (7-days later). Ninety-seven completed the study. History of 'taking time off' work/study because of pericoronitis during the past year was significantly associated with preoperative OHIP-14 and OHQoL-UK scores. There were significant differences in SF-12, OHIP-14, OHQoL-UK scores during the immediate postoperative period compared with preoperatively, when postoperative symptoms were prevalent. At the review appointment, OHIP-14 and OHQoL-UKscores were associated with clinical findings. The measures were valid and sensate in relation to oral surgery. However, those specific to oral health were more discerning than the general scale. PMID- 12576041 TI - A self-drilling intermaxillary fixation screw. PMID- 12576042 TI - The custom-made titanium orbital floor prosthesis in reconstruction for orbital floor fractures. PMID- 12576043 TI - Use of Hirudo medicinalis by maxillofacial surgical units in the United Kingdom: current views and practice. PMID- 12576044 TI - Epidural spinal abscess following dental extraction--a rare and potentially fatal complication. PMID- 12576045 TI - Gingival metastasis of malignant mesenchymal tumour in the thigh. PMID- 12576046 TI - Posterior scleritis--an unusual presentation of facial pain. AB - Defining the aetiology of facial pain can be difficult. While acknowledging the principle of 'common things are common', it is prudent to consider all possible causes. We describe a patient with posterior scleritis that presented as facial pain and was cured by oral prednisolone. PMID- 12576047 TI - An unusual parotid lump. PMID- 12576048 TI - Re: traumatic fracture pattern following mandibular ramus osteotomy. PMID- 12576049 TI - Re: immediate reconstruction after maxillectomy. Br J Oral Maxillofac Surg 2002; 40:183-190. PMID- 12576051 TI - Isolation of a caveolae-enriched fraction from rat lung by affinity partitioning and sucrose gradient centrifugation. AB - Caveolae were isolated from rat lungs by a combination of affinity partitioning and sucrose gradient centrifugation. After homogenization of the lungs directly in a polyethylene glycol-dextran two-phase system and conventional phase partitioning, the polyethylene glycol-rich top phase was affinity partitioned with fresh bottom phase containing dextran-linked wheat-germ agglutinin. The lectin selectively attracted plasma membranes to the bottom phase. The isolated plasma membrane fraction was treated with Triton X-100 or, alternatively, sonicated before centrifugation in a stepwise sucrose gradient. Caveolin-enriched material collected at the 5/24% sucrose boundary. This material also contained 5' nucleotidase activity and actin. Electron microscopy showed the material to consist of a homogeneous population of 50- to 100-nm vesicles. This purification protocol should allow the facile purification of caveolae also from other tissues, facilitating structural and functional studies. PMID- 12576052 TI - A quantitative method for measuring protein phosphorylation. AB - We have developed a novel method for quantitating protein phosphorylation by a variety of protein kinases. It can be used with purified kinases and their substrates in vitro or in combination with cell extracts. The method is based on the knowledge that protein kinase C (PKC) adds three phosphates to each molecule of its preferred substrate, myelin basic protein (MBP). A time course is performed in which a kinase is allowed to phosphorylate its preferred substrate or the protein under investigation in the presence of [gamma-32P]ATP. At the same time PKC is allowed to fully phosphorylate MBP. After resolving the products by SDS-PAGE, electrophoretic transfer, and determining the degree of incorporation of 32P by phosphorImager analysis, the data are converted to moles phosphate/mole protein by normalization with phosphorylated MBP. The method is both sensitive and relatively rapid and all the steps are commonly available in the biochemistry laboratory. We have used this method to confirm and extend information on the relationship of MEK1 and MAPK/Erk2 in rat lung fibroblasts exposed to V(2)O(5). A 4-h exposure to V(2)O(5) results in partial phosphorylation of MAPK/Erk2 such that 25% of the potential phosphorylation sites are occupied. We also demonstrate that despite multiple potential phosphorylation sites, recombinant human AP endonuclease is weakly phosphorylated in vitro (4% at best) by PKC, cGMP dependent protein kinase, casein kinase II, and casein kinase I and not at all phosphorylated by MAPK. Furthermore we are unable to demonstrate phosphorylation in cell extracts from HeLa cells, mouse fibroblasts after oxidative damage with H(2)O(2) or alkylation damage with methylmethane sulfonate, or rat lung fibroblasts after oxidative damage with V(2)O(5). PMID- 12576053 TI - Determination of peptides and amino acids from wool and beer with sensitive fluorescent reagent 2-(9-carbazole)-ethyl chloroformate by reverse phase high performance liquid chromotography and liquid chromotography mass spectrometry. AB - A new method for the sensitive determination of amino acids and peptides using the tagging reagent 2-(9-carbazole)-ethyl chloroformate (CEOC) with fluorescence (FL) detection has been developed. Identification of derivatives was carried out by liquid chromotography mass spectrometry. The chromophore in the 2-(9 fluorenyl)-ethyl chloroformate (FMOC) reagent was replaced by carbazole, which resulted in a sensitive fluorescence lerivatizing agent CEOC. CEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. Studies on derivatization demonstrate excellent derivative yields over the pH range 8.8 10.0. Maximal yields close to 100% are observed with three- to fourfold molar reagent excess. Derivatives exhibit strong fluorescence and allow direct injection of the reaction mixture with no significant disturbance from the major fluorescent reagent degradation by-products, such as 2-(9-carbazole)-ethanol and bis-(2-(9-carbazole)-ethyl) carbonate. In addition, the detection responses for CEOC derivatives are compared to those obtained with FMOC. The ratios AC(CEOC)/AC(FMOC) = 1.00-1.82 for fluorescence (FL) response and AC'(CEOC)/AC'(FMOC) = 1.00-1.21 for ultraviolet (UV) response are observed (here, AC and AC' are, respectively, FL and UV response). Separation of the derivatized peptides and amino acids has been optimized on a Hypersil BDS C18 column. Excellent linear responses are observed. This method was used successfully to analyze protein hydrolysates from wool and from direct-derivatized beer. PMID- 12576054 TI - Simultaneous determination of methionine sulfoxide and methionine in blood plasma using gas chromatography-mass spectrometry. AB - Methionine sulfoxide is an oxidation product of methionine with reactive oxygen species via 2-electron-dependent mechanism. Such oxidants can be generated from activated neutrophils; therefore, methionine sulfoxide can be regarded as a biomarker of oxidative stress in vivo. We describe here a method for the simultaneous determination of methionine sulfoxide and methionine in blood plasma using gas chromatography-mass spectrometry with isotopically labeled compounds as internal standards. This method comprises the inclusion of [Me-13C, Me 2H(3)]methionine sulfoxide and [Me-13C, Me-2H(3)]methionine into plasma, the removal of plasma proteins using acetonitrile, the purification of amino acids with cation-exchange chromatography, and the derivatization of methionine sulfoxide and methionine to their corresponding tert-butyldimethylsilyl derivatives using N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide. Quantitation was performed by electron impact mode. The levels of methionine sulfoxide in healthy human blood plasma were 4.0 +/- 1.0 microM (means +/- SD, n = 8), indicating that approximately 10% of methionine is detected as the oxidized form in healthy human plasma. The ratio of methionine sulfoxide in total methionine increased with treatment of human blood with phorbol 12-myristate 13 acetate, while this ratio remained constant in plasma from alloxan-induced hyperglycemic rabbits. These results indicate that this method is applicable for plasma samples and methionine sulfoxide can represent oxidative stress caused by nonradical oxidation in vivo. PMID- 12576055 TI - Capillary electrophoretic enzyme immunoassay with electrochemical detection for thyroxine. AB - A capillary electrophoretic enzyme immunoassay with electrochemical detection (CE EIA-ED) has been developed. In this method, antigen (Ag) competes with horseradish peroxidase (HRP)-labeled antigen (HRP-Ag) for a limited number of antibody (Ab) binding sites. The free HRP-Ag and the bound HRP-Ag-Ab complex are separated by capillary electrophoresis in a separation capillary. Then they catalyze the oxidation of their enzyme substrate 3,3',5,5'-tetramethylbenzide (TMB (reduced form)) with H(2)O(2) in a reaction capillary, which follows the separation capillary. The reaction product (TMB (oxidized form)) is amperometrically determined using a carbon fiber microdisk bundle electrode at the outlet of the reaction capillary. Due to the amplification of the enzyme, the concentration of TMB(Ox) is much higher than those of free HRP-Ag and the bound HRP-Ag-Ab complex. Therefore, the limit of detection (LOD) of CE-EIA-ED is very low. The method has been used to determine thyroxine in human serum. A concentration of LOD of 3.8 x 10(-9)mol/L, which corresponds to a mass LOD of 23.2 amol, was achieved. PMID- 12576056 TI - Protein immobilization on plasma-polymerized ethylenediamine-coated glass slides. AB - For protein chip construction, protein immobilization on the surface of the glass slide is essential. It was previously reported that glass slides are embedded with chemicals that contain primary amines and aldehydes for protein immobilization. We fabricated a plasma-polymerized ethylenediamine (PPEDA)-coated slide that exposed primary amines. For the plasma polymer deposition on the glass slide, the inductively coupled plasma (ICP) power was found to be a critical factor in sustaining a high density of amine on the surface of the PPEDA films. We prepared PPEDA-coated slides at three different ICP powers (3, 30, or 70 W). In the slide that was prepared at a low ICP power (3 W), we detected a high density of primary amine. Therefore, the fluorescein isothiocyanate-conjugated immunoglobulin G (IgG) was highly immobilized to the PPEDA-coated slide that was prepared at the low ICP power. For protein immobilization, 1-ethyl-3-[3 dimethylaminopropyl]carbodiimide hydrochloride (EDC) was used as a cross-linker. The immobilization of the protein to the PPEDA-coated slide was carried out by consecutive incubations with 1 mg/ml EDC for 5 min and 0.1 mg/ml IgG for 1 h. This efficiently produced the functionally active protein-immobilized slide. Therefore, this work shows that the plasma technique can be applied to produce a high-quality glass surface for the immobilization of proteins and other materials. PMID- 12576057 TI - In situ assay of the intramitochondrial enzymes: use of alamethicin for permeabilization of mitochondria. AB - The channel-forming antibiotic alamethicin was used to permeabilize mitochondrial membranes for the low molecular mass hydrophilic substrates NADH and ATP. Alamethicin-treated mitochondria show high rotenone-sensitive NADH oxidase, NADH quinone reductase, and oligomycin-sensitive and carboxyatractylate-insensitive ATPase activities. Alamethicin does not affect Complex I and ATPase activities in inside-out submitochondrial particles. Permeabilized mitochondria quantitatively retain their aconitase and iso-citrate dehydrogenase activities. Electron microscopy of alamethicin-treated mitochondria reveals no disruption of their outer and inner membranes. From the results obtained it is recommended, that alamethicin be used for the in situ catalytic assay of intramitochondrially located enzymes. PMID- 12576058 TI - Versatile method employing basic techniques of genetic engineering to study the ability of low-molecular-weight compounds to bind covalently with DNA in cell free systems. AB - Numerous antitumor and carcinogenic compounds and free radicals are able to modify DNA by forming covalent bonds, mainly with nucleophilic centers in nucleobases. Such a binding is usually of utmost importance for the biological outcome. The level of DNA adducts formed by a given agent is in most cases extremely low; hence their detection is very difficult. Here we propose a simple approach, exploiting techniques widely used in genetic engineering, to demonstrate and characterize the covalent modification of a DNA fragment by any low-molecular-weight compound of interest in a cell-free system. The specifically designed, several-hundred-base-pairs-long double-stranded deoxyoligonucleotide (PCR amplified)--subject to modification--includes two restriction sites: one containing only GC base pairs recognized by restriction endonuclease MspI and the other including only AT base pairs recognized by restriction endonuclease Tru1I. The covalent modification of the restriction sites abolishes their recognition and thus cleavage by the endonucleases applied. The formation of DNA adducts is induced by incubating the oligonucleotide with increasing concentrations of a studied compound, in the appropriate activating system if required. Then, the modified oligonucleotide is submitted to digestion by the above-mentioned restriction endonucleases and the DNA fragments are separated by polyacrylamide gel electrophoresis. The inhibition of cleavage indicates the occurrence of covalent modification of the restriction site(s) while simultaneously pointing at the kind of base pairs involved in DNA adduct formation. The validation of the method was performed for two DNA binding antitumor compounds, cisplatin and CC 1065, which form adducts preferentially with guanine and adenine, respectively. PMID- 12576059 TI - Further development of an electroosmotic medium pump system for preparative disk gel electrophoresis. AB - A simple and practical 6.8-cm-diameter (36.30-cm(2) cross-sectional-area) preparative disk gel electrophoresis device, based on the design of M. Hayakawa et al. (Anal. Biochem. 288 (2001) 168), in which the elution buffer is driven by an electroosmotic buffer flow through the membrane into the elution chamber from the anode chamber was constructed. We have found that the dialysis membranes employed provide suitable flow rates for the elution buffer, similar to those of an earlier 3.6-cm-diameter device, resulting in the prevention of excess eluate dilution. The efficiency of this device was demonstrated by the fractionation of a bovine serum albumin (BSA) Cohn V fraction into monomer, dimer, and oligomer components using nondenaturing polyacrylamide gel electrophoresis (native-PAGE). The maximum protein concentration of the eluate achieved was 133 mg/ml of BSA monomer, which required a dilution of the eluate for subsequent analytical PAGE performance. As a practical example, the two-dimensional fractionation of soluble dipeptidyl peptidase IV (sDPP IV) from 50 ml fetal bovine serum (3.20 g protein) per gel is presented. The sDPP IV enzyme protein was recovered in a relatively short time, utilizing a 6.5% T native-PAGE and subsequential sodium dodecyl sulfate-PAGE system. This device enhances the possibility of continuous electrophoretic fractionation of complex protein mixtures on a preparative scale. PMID- 12576060 TI - Homogeneous assay for biotin based on Aequorea victoria bioluminescence resonance energy transfer system. AB - Here we describe a homogeneous assay for biotin based on bioluminescence resonance energy transfer (BRET) between aequorin and enhanced green fluorescent protein (EGFP). The fusions of aequorin with streptavidin (SAV) and EGFP with biotin carboxyl carrier protein (BCCP) were purified after expression of the corresponding genes in Escherichia coli cells. Association of SAV-aequorin and BCCP-EGFP fusions was followed by BRET between aequorin (donor) and EGFP (acceptor), resulting in significantly increasing 510 nm and decreasing 470 nm bioluminescence intensity. It was shown that free biotin inhibited BRET due to its competition with BCCP-EGFP for binding to SAV-aequorin. These properties were exploited to demonstrate competitive homogeneous BRET assay for biotin. PMID- 12576061 TI - Time-minimized determination of ribosome and tRNA levels in bacterial cells using flow field-flow fractionation. AB - The evaluation of the translation capacity of cells that produce recombinant proteins can be made by monitoring their ribosomal composition. In a previous use of asymmetrical flow field-flow fractionation (AsFlFFF) for this purpose the overall analysis time was more than 1 h and 40 min, based on a standard protocol for cell harvest, washing, cell disruption, and the final 8-min AsFlFFF determination of ribosome and subunits. In the present work the overall analysis time was reduced to 16 min. The washing step was deleted and a time-consuming freeze-thaw cycle. Cell disruption was obtained by a time-minimized lysozyme and detergent treatment for 1.5 min, respectively. The ribosomal material was finally fractionated and quantified in only 6 min, without previous centrifugation, using AsFlFFF. The great time reduction will enable the future use of AsFlFFF at-line to a growing cell cultivation, continuously monitoring the change in ribosomal composition or in other applications requiring high sample throughput. To demonstrate the high efficiency of the method the ribosome and tRNA composition in an Escherichia coli cultivation was monitored every half an hour, giving 18 measurements across the complete growth curve, a frequency of data enough to make decisions about induction or termination of the cultivation. PMID- 12576062 TI - Equilibrium titration of protein-ligand binding affinity in the presence of volatile reagents--a semiautomated approach. AB - An existing method of equilibrium titration was significantly improved for investigating the effects of volatile anesthetics on Ca(2+) binding characteristics of human recombinant cardiac troponin C in in vitro conditions. The modified method increases stability of volatile compound concentrations in solution and allows for faster and more accurate data acquisition. The time to complete a titration series could be reduced from 28.3 +/- 6.2 min to 9.3 +/- 2.1 min, whereas the dispersion for pK(d) was decreased from 2.16 +/- 0.27 to 0.63 +/ 0.27. The method utilizes a semiautomatic approach to continuously monitor stability of fluorescence signals in a sealed chamber with greatly reduced air space. PMID- 12576063 TI - Picomolar analysis of flavins in biological samples by dynamic pH junction sweeping capillary electrophoresis with laser-induced fluorescence detection. AB - Sensitive capillary electrophoresis (CE) methods are required for emerging areas of biochemical research such as the metabolome. In this report, dynamic pH junction-sweeping CE with laser-induced fluorescence (LIF) detection is applied as a robust single method to analyze trace amounts of three flavin derivatives, riboflavin, flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD), from several types of samples including bacterial cell extracts, recombinant protein, and biological fluids. Submicromolar amounts of flavin coenzymes were measured directly from formic acid cell extracts of Bacillus subtilis. Significant differences in flavin concentration were measured in cell extracts derived from either glucose or malate as the carbon source in the culture media. Quantitative assessment of FAD and FMN content from selected flavoenzymes was demonstrated after heat denaturation to release noncovalently bound coenzymes and deproteinization. This method was also applied to the analysis of free flavins in pooled human plasma and urine without the need for laborious off-line sample preconcentration. Picomolar detectability of flavins by CE-LIF detection was realized with on-line preconcentration (up to 15% capillary length used for injection) by dynamic pH junction-sweeping, resulting in a limit of detection (S/N = 3) of about 4.0 pM for FAD and FMN. This represents over a 60-fold improvement in concentration sensitivity compared to those of previous techniques using conventional injections. The method was validated in terms of reproducibility, sensitivity, linearity, and specificity. Flavin analysis by dynamic pH junction-sweeping CE-LIF offers a simple, yet sensitive way to analyze trace levels of flavin metabolites from complex biological samples. PMID- 12576064 TI - Determination of prostate specific antigen mRNA in peripheral blood by reverse transcriptase polymerase chain reaction and a simple chemiluminometric hybridization assay in a high-throughput format. AB - In recent years, the mRNA for prostate-specific antigen (PSA) has been investigated as a potential marker for molecular staging of prostate cancer. We report a simple, rapid, and sensitive assay protocol for the quantification of PSA mRNA in peripheral blood by using reverse transcriptase polymerase chain reaction (RT-PCR) and a chemiluminometric hybridization assay. A recombinant RNA internal standard (IS) that has the same size and primer binding sites as the PSA mRNA is included in the RT-PCR mixture. Total RNA from the sample is coextracted with a constant amount of IS RNA and subjected to RT-PCR. Amplified sequences are labeled with biotin during PCR by using a biotinylated upstream primer. The products are heat-denatured and hybridized with oligonucleotide-specific probes (for PSA and IS) that are immobilized in microtiter wells. Immobilization of oligonucleotide probes is achieved by adsorption of their conjugates with bovine serum albumin. The hybrids are measured using alkaline phosphatase-labeled streptavidin and a dioxetane chemiluminogenic substrate. The ratio of the luminescence values obtained for the PSA mRNA and the RNA IS is a linear function of the initial amount of PSA mRNA present in the sample prior to RT-PCR amplification. The linear range extended from 50 to 500,000 PSA mRNA copies, and the overall reproducibility of the assay, including RT-PCR and hybridization, ranged from 11.5 to 14.2%. Samples containing total RNA from PSA-expressing LNCaP cells give luminescence ratios that are linearly related to the number of cells in the range of 0.04-400 cells. The method was applied to PSA mRNA determination in peripheral blood of healthy individuals, patients with benign prostate hyperplasia, patients with prostate cancer, and patients with other types of localized cancer. PMID- 12576065 TI - Simultaneous resolution of underivatized regioisomers and stereoisomers of arachidonate epoxides by capillary electrophoresis. AB - cis-Epoxyeicosatrienoic acids (EETs) and their hydrolysis products (threo-DHETs) have been proposed to be endothelial-dependent hyperpolarizing factors (EDHFs) which upregulate blood flow when tissue perfusion is impaired. Various EET regioisomers and enantiomers are formed from arachidonate by inducible cytochrome P450 epoxygenase isoforms, and tissue EET profiles may vary with diet, time, and disease. Because EET actions and metabolism may be regio- and stereospecific, convenient methods to measure profiles of EET isomers in tissues are needed. In the current studies, we describe two simple capillary electrophoretic methods for resolving EETs. The first method involves capillary electrophoresis with a mixture of neutral and anionic beta-cyclodextrins, which in one step baseline resolves underivatized EET regioisomers and their enantiomers. Low picogram amounts of EET enantiomers were identified based on migration times and UV spectra. The method was also used to assess the antipode purity of EET standards, and to determine murine hepatic levels of EET enantiomers. The second method involves capillary electrochromatography, which also baseline-resolves underivatized EET and DHET regioisomers in one step. We conclude that in EET assays the major advantages of capillary electrophoresis over reversed-phase HPLC are improved peak efficiency, sensitivity, and resolution, plus precise coelution of deuterated and nondeuterated EETs. PMID- 12576066 TI - Affinity extraction combined with stable isotope dilution LC/MS for the determination of 5-methyltetrahydrofolate in human plasma. AB - The predominant circulating folate monoglutamate in human plasma (>90%), and thus the most significant folate for accurately diagnosing folate deficiency, is 5 methyltetrahydrofolic acid (5 MT). Folate deficiency is typically indicated when circulating folate levels are < or = 3 ng/mL. The quantitative determination of plasma folates in general, and of 5 MT in particular, is complicated by their naturally low levels (pg/mL to ng/mL), their instability, and their tendency to interconvert. Highly specific and sensitive analytical methods are needed to accurately quantify endogenous 5 MT in human plasma. A method that utilizes the specific high-affinity binding sites of bovine folate binding protein (FBP) and the selectivity and sensitivity of selected ion monitoring mode isotope-dilution liquid chromatography/mass spectrometry (LC/MS) to quantify plasma 5 MT has been developed. The method is based on the solid-phase affinity extraction (SPAE) of 5 MT and its stable isotopically labeled analogue ([13C(5)]5 MT) from plasma (1 mL) using FBP immobilized to polymeric beads. The excess high-affinity binding sites on the affinity columns enable quantitative extraction of 5 MT from plasma under optimized sample pH conditions. Additionally, it is demonstrated that plasma proteins do not hinder the determination of 5 MT; therefore, protein precipitation is not required before the affinity extraction step. Detection and quantification of the extracted 5 MT is provided by positive-ion mode LC/MS in which the protonated molecular ions [M+H](+) of the analyte and the internal standard are monitored. The method shows linearity over three orders of magnitude (0.04-40 ng/mL) and has limits of detection and quantification of 0.04 and 0.4 ng/mL, respectively. Calibration curves obtained by spiking 5 MT into plasma exhibited good linearity between 0 and 25 ng/mL and both the plasma calibration standards and the plasma samples were stable for at least 48 h at room temperature. The recovery (average +/- % RSD) of 5 MT spiked into plasma from 5 to 25 ng/mL was 98.0% +/- 1.6% (n = 15). 5 MT levels determined by SPAE-LC/MS compared to "total folate" levels determined by radioassay and microbiological assay were discordant. Reasons for the discordancy are theorized, but it is clear that there exists an urgent need for clinical reference materials containing certified folate levels. PMID- 12576067 TI - High-performance liquid chromatography purification of 26-bp serial analysis of gene expression ditags results in higher yields, longer concatemers, and substantial time savings. AB - In contrast to DNA chips, serial analysis of gene expression (SAGE) is not dependent on genes having been previously identified for their monitoring. Although useful, the method can be technically challenging, and particularly the last steps including concatenation and cloning may result in less than optimal results. We propose that many of the encountered problems can be attributed to the purification of the 26-bp ditags by polyacrylamide gel electrophoresis. Low yields, gel contaminants, potential exposure to degrading enzymes during handling and lengthy separation all disfavor the method. We introduce purification of 26 bp ditags by reverse-phase high-performance liquid chromatography (HPLC) using polystyrene/divinylbenzene columns and tetraethylammonium acetate buffer with acetonitrile as mobile phase. The method is fast and gives excellent results. Ditags purified by HPLC readily ligate to high-molecular-weight concatemers leading to their efficient cloning. The method should substantially facilitate the construction of SAGE libraries. PMID- 12576068 TI - Effect of large refractive index gradients on the performance of absorption optics in the Beckman XL-A/I analytical ultracentrifuge: an experimental study. AB - An analytical centrifuge cell was modified to detect refraction of light transmitted through the cell caused by refractive index gradients formed by sedimenting solute during centrifugation. Sedimentation velocity and sedimentation equilibrium experiments were carried out in this cell on solutions containing high concentrations of protein and polysaccharide in a Beckman-Coulter XLA analytical ultracentrifuge. Analysis of the results indicates that in the absence of an optical artifact easily recognized as a "black band," the dependence of apparent absorbance upon radial position reported by the instrument may be considered a reliable measure of the solute concentration gradient. PMID- 12576069 TI - The use of lymphocytes to screen for oxidative phosphorylation disorders. AB - Biochemical analysis of oxidative phosphorylation (OXPHOS) disorders is traditionally carried out on muscle biopsies, cultured fibroblasts, and transformed lymphocytes. Here we present a new screening technique using lymphocytes to identify OXPHOS dysfunction and initially avoid an invasive diagnostic procedure. Lymphocytes represent an easily obtainable source of tissue that presents advantages over the use of fibroblasts or lymphoblast cell lines. The time delay in culturing skin fibroblasts and the interactions between cell transformation and mitochondrial activity are avoided in this methodology. The method requires a small amount of blood (<5 mL); can be completed in a few hours, and allows for repeated measurements. Our assay has been adapted from published methods utilizing cultured fibroblasts and transformed lymphocytes, and our data suggest that measurement of ATP synthesis in lymphocytes is an effective screening tool for diagnosing OXPHOS disorders. This method may also provide an objective tool for monitoring response to treatment and evaluating progression of disease. PMID- 12576070 TI - Global metabolite analysis: the influence of extraction methodology on metabolome profiles of Escherichia coli. AB - The global pool of all metabolites in a cell, or metabolome, is a reflection of all the metabolic functions of an organism under any particular growth condition. In the absence of in situ methods capable of universally measuring metabolite pools, intracellular metabolite measurements need to be performed in vitro after extraction. In the past, a variety of cell lysis methods were adopted for assays of individual metabolites or groups of intermediates in pathways. In this study, metabolites were extracted from Escherichia coli using six different commonly used procedures including acid or alkaline treatments, permeabilization by freezing with methanol, high-temperature extraction in the presence of ethanol or methanol, and by lysis with chloroform-methanol. Metabolites were extracted by the six methods from cells grown under identical conditions and labeled with [14C]glucose. The metabolomes were compared after 2-dimensional thin-layer chromatography of labeled compounds. For global analysis, extraction with cold ( 40 degrees C) methanol showed the greatest promise, allowing simultaneous resolution of more than 95 metabolite spots. In contrast, 80 or less spots were obtained with other extraction methods. Extraction also influenced quantitative analysis of particular compounds. Metabolites such as adenosine exhibited up to 20-fold higher abundance after cold methanol extraction than after extraction with acid, alkali, or chloroform. The simplicity, rapidity, and universality of cold methanol extraction offer great promise if a single method of lysis is to be adopted in metabolome analysis. PMID- 12576071 TI - A single-step procedure for the synthesis of photoreactive and radioactive glycerolipids. AB - A single-step procedure was developed for the incorporation of iodoazide into oleic acid, triolein, and phosphatidylcholine. Iodoazide was generated using [125I]iodomonochloride and sodium azide that was found to add stereospecifically in a variety of olefins. Photoreactive and radiolabeled triacylglycerol and phosphatidylcholine were synthesized with a moderate yield and high specific activity. The stability of both the radiolabel and the photoreactive group was studied under ultraviolet light under aqueous as well as anhydrous conditions. These synthesized analogs act as substrates in the dark, and as irreversible inhibitors under ultraviolet irradiation for the target hydrolytic enzymes. The synthesized radiolabeled photoprobes were subsequently used to label lipase and phospholipase A(2). The results highlighted the efficiency and rapidity of the method and its potential application in the study of lipid-metabolizing enzymes. PMID- 12576072 TI - Detection and analysis of enzymatic DNA methylation of oligonucleotide substrates by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) mass spectrometry was employed to analyze DNA methylation carried out by the Escherichia coli dam DNA methyltransferase using oligonucleotide substrates with molecular masses of 5000-10,000 Da per strand. The mass spectrometry assay offers several advantages: (i) it directly shows the methylation as the increase in the mass of the substrate DNA, (ii) it is nonradioactive, (iii) it is quantitative, and (iv) it can be automated for high throughput applications. Since unmethylated and methylated DNA are detected, the ratio of methylation can be determined directly and accurately. Furthermore, the assay allows detection individually of the methylation of several substrates in competition, offering an ideal setup to analyze the specificity of DNA interacting with enzymes. We could not identify methylation at any noncanonical site, indicating that the dam MTase is a very specific enzyme. Finally, MALDI-TOF mass spectrometry permitted assessment of the number of methyl groups incorporated into each DNA strand, thereby, allowing study of mechanistic details such as the processivity of the methylation reaction. We provide evidence that the dam MTase modifies DNA in a processive reaction, confirming earlier findings. PMID- 12576073 TI - Photometric determination of proline in quartz microplates: remarks on specificity. PMID- 12576074 TI - Modification of Job's method for determining the stoichiometry of protein-protein complexes. PMID- 12576075 TI - Complete separation of adenine nucleotides for ATPase activity assay by ion-pair reversed-phase high-performance liquid chromatography. PMID- 12576076 TI - Hydroxyapatite chromatography: altering the phosphate-dependent elution profile of protein as a function of pH. PMID- 12576077 TI - A radiochemical pyruvate dehydrogenase assay: activity in heart. PMID- 12576078 TI - Oligonucleotide analysis by sequential injection before analysis (SIBA) capillary electrophoresis. PMID- 12576079 TI - Binding of cationic cell-permeable peptides to plastic and glass. AB - Cell-penetrating peptides derived from hydrophilic regions of the homeoprotein Antennapedia (Antp) or the transcription-regulating factor Tat have been used to transport several peptide and oligonucleotide cargoes into the interior of cells. Such vector peptides penetrate cells, in part, because they contain multiple lysine and arginine residues. Using radiolabeled peptide cargoes covalently linked to Antp- or Tat-related vectors, or to D-Arg heptamers, we found that a significant amount of the label remained tightly bound to plastic and glass surfaces. Binding of the labeled conjugates was due entirely to the cationic vector moieties. Under certain conditions, such non-specific binding could be mistaken for cellular penetration. PMID- 12576081 TI - Identification of a vasopressin-like immunoreactive substance in hydra. AB - Vasopressin (VP)-like immunoreactivity has long been known in the hydra nervous system, but has not yet been structurally identified. In this study, using HPLC fractionation and an immunological assay, we have purified two peptides, FPQSFLPRGamide and SFLPRGamide, from Hydra magnipapillata. Both the peptides shared the same C-terminal structure, -PRGamide, with Arg-VP. The nonapeptide proved to be Hym-355, a peptide that stimulates neuronal differentiation in hydra. Detailed evaluation by competitive enzyme-linked immunosorbent assay (ELISA) and double immunostaining using anti-VP and anti-Hym-355 antibodies enabled us to conclude that the two peptides account for a major part of the VP like immunoreactivity in hydra nerve cells. PMID- 12576080 TI - Purification and characterization of alpha- and beta-benincasins, arginine/glutamate-rich peptides with translation-inhibiting activity from wax gourd seeds. AB - Two peptides, with a molecular mass of about 11 kDa and an N-terminal sequence abundant in arginine and glutamine residues, were isolated from wax gourd seeds. The isolation protocol included affinity chromatography on Affi-gel blue gel, ion exchange chromatography on Mono-S and gel filtration on Superdex 75. The peptides, designated alpha- and beta-benincasins, inhibited cell-free translation in a rabbit reticulocyte lysate system with an IC(50) of 20 and 320 pM, respectively. Alpha-benincasin exhibited weak antifungal activity toward Coprinus comatus and Physalospora piricola but not toward Mycosphaerella arachidicola. PMID- 12576082 TI - Isolation, identification and localization of a second beetle antidiuretic peptide. AB - We isolated from head extracts of Tenebrio molitor a peptide that inhibits fluid secretion by the Malpighian tubules of this insect. This second antidiuretic factor, ADFb, like the previously published ADFa, works through cyclic GMP as a second messenger. It has primary structure Tyr-Asp-Asp-Gly-Ser-Tyr-Lys-Pro-His Ile-Tyr-Gly-Phe-OH with an EC(50) of approximately 240 pM in a fluid secretion assay. This peptide is now the second sequenced endogenous insect ADF which inhibits Malpighian tubule fluid secretion. Immunohistochemical techniques show that the peptide is localized in the brain; it appears to be produced mainly in two pairs of bilaterally symmetrical cells in the protocerebrum. PMID- 12576083 TI - Isolation and characterization of novel tachykinins from the posterior salivary gland of the common octopus Octopus vulgaris. AB - Two novel tachykinins (OctTK-I: Lys-Pro-Pro-Ser-Ser-Ser-Glu-Phe-Ile-Gly-Leu-Met NH(2) and OctTK-II: Lys-Pro-Pro-Ser-Ser-Ser-Glu-Phe-Val-Gly-Leu-Met-NH(2)) were isolated from the posterior salivary gland of the octopus (Octopus vulgaris) using a contraction assay of the carp rectum. These peptides had in common the pentapeptide sequence -Phe-X-Gly-Leu-Met-NH(2) at the C-terminal and induced immediate contractions on the carp rectum and the guinea-pig ileum. cDNAs encoding their precursor proteins were cloned. The OctTK gene was expressed in the posterior salivary gland and the expression was localized in mucus-secreting cells of the gland. The results suggested that OctTKs might be secreted as a venomous substance acting on vertebrates such as fishes, which are the prey or natural enemies of the octopus. PMID- 12576084 TI - Deamidase inactivates a D-amino acid-containing Aplysia neuropeptide. AB - Membrane-catalyzed degradation of the cardioexcitatory peptide, Asn-D-Trp-Phe NH(2) (N(d)WF-NH(2)), which was previously isolated from Aplysia, was investigated in relation to its inactivation mechanism. The principal degradation was deamidation of the C-terminal amide, producing biologically inert Asn-D-Trp Phe-OH (N(d)WF-OH). Among membrane fractions prepared from different organs, the fraction from the ganglia showed the highest specific activity of the deamidation reaction. The deamidase activity was inhibited by Ebelactone B and the serine protease inhibitor, phenylmethanesulfonyl fluoride (PMSF), while the degradation of the synthetic stereoisomer, Asn-Trp-Phe-NH(2) (N(l)WF-NH(2)), was sensitive to the divalent cation-chelating agent, o-phenanthroline, and aminopeptidase inhibitors, amastatin and bestatin. The presence of D-Trp residue in the second position of N(d)WF-NH(2) endows this peptide not only with stereospecific bioactivity but also peptidase stability. The deamidation reaction seems to be the major inactivation mechanism for this peptide. PMID- 12576085 TI - Somatostatin-like receptors in goldfish: cloning of four new receptors. AB - In this study, four somatostatin-like receptor (Sst) cDNAs were identified from goldfish pituitary, using RT-PCR screening and rapid amplification of cDNA ends (RACE) strategies. These include two type-five like Sst (Sst(5B) and Sst(5C)) and two type-three like Sst receptors (Sst(3A) and Sst(3B)), designated based on their amino acid sequence similarities to the known mammalian and fish Sst(5) and Sst(3). Both Sst(5C) and Sst(3A) mRNAs are widely expressed in all brain regions and pituitary; however, Sst(3B) expression is restricted to forebrain and Sst(5B) expression is mainly detected in pituitary and spinal cord. PMID- 12576086 TI - Evidence for the existence of a new receptor for CGRP, which is not CRLR. AB - Receptors for calcitonin gene-related peptide (CGRP), a neuropeptide known to be the most potent vasodilator, are abundantly expressed in cerebellum. A monoclonal antibody to cerebellar CGRP receptors specifically detects a 66 kDa protein from rat cerebellum and other rat and human tissues, but not from SK-N-MC cells which express calcitonin receptor-like receptor (CRLR), a recently described component of CGRP receptors. In contrast, mRNA expression for CRLR was abundant in SK-N-MC cells, but it was undetectable in rat cerebellum. Furthermore, the antibody could not detect any immunoreactive protein in HEK 293 cells transiently transfected with CRLR and receptor activity-modifying protein 1 (RAMP(1)) indicating the possible existence of another CGRP receptor, which does not involve CRLR. Due to the absence of biochemical or structural data on the existence of a CGRP(2) receptor and the new data provided in this paper, we suggest to identify the two CGRP receptors as CGRP-A and CGRP-B. PMID- 12576087 TI - Structure-activity relationships of the melanocortin tetrapeptide Ac-His-DPhe-Arg Trp-NH2 at the mouse melanocortin receptors. Part 3: modifications at the Arg position. AB - The melanocortin pathway is involved in the regulation of several physiological functions including skin pigmentation, steroidogenesis, obesity, energy homeostasis, and exocrine gland function. This melanocortin pathway consists of five known G-protein coupled receptors, endogenous agonists derived from the proopiomelanocortin (POMC) gene transcript, the endogenous antagonists Agouti and the Agouti-related protein (AGRP) and signals through the intracellular cAMP signal transduction pathway. The melanocortin-3 receptor (MC3R) and melanocortin 4 receptor (MC4R) located in the brain are implicated as participating in the metabolic and food intake aspects of energy homeostasis and are stimulated by melanocortin agonists such as alpha-melanocyte stimulation hormone (alpha-MSH). All the endogenous (POMC-derived) melanocortin agonists contain the putative message sequence "His-Phe-Arg-Trp." Herein, we report 12 tetrapeptides, based upon the template Ac-His(6)-DPhe(7)-Arg(8)-Trp(9)-NH(2) (alpha-MSH numbering) that have been modified at the Arg(8) position by neutral, basic, or acidic amino acid side chains. These peptides have been pharmacologically characterized for agonist activity at the mouse melanocortin receptors MC1R, MC3R, MC4R, and MC5R. The most notable results of this study include the observation that removal of the guanidinyl side chain moiety results in decreased melanocortin receptor potency, but that this Arg(8) side chain is not critical for melanocortin receptor agonist activity. Additionally, incorporation of the homoArg(8) residue results in 56-fold MC4R versus MC3R selectivity, and the Orn(8) residue results in 123-fold MC4R versus MC5R and 63-fold MC5R versus MC3R selectivity. PMID- 12576088 TI - Bombesin receptor subtype-3 modulates plasma insulin concentration. AB - Mice lacking a functional bombesin receptor subtype-3 (BRS-3) develop mild obesity. However, the origin of obesity in BRS-3 knockout (KO) mice remains unclear. We used a strain-crossing strategy to investigate the physiological role of the BRS-3 pathway. We crossed female heterozygous BRS-3 KO mice (X-/X) and male KK-Ay mice (Ay/+) to obtain BRS-3 KO/KK-Ay hybrid animals. In X-/Y:Ay/+ mice, plasma insulin concentrations were significantly higher, and on the oral glucose tolerance test, the additional secretion of insulin was impaired compared to other genotypes. Our results indicate that the BRS-3 pathway contributes to the regulation of plasma insulin concentrations. PMID- 12576089 TI - Endogenous amylin contributes to the anorectic effects of cholecystokinin and bombesin. AB - Previous studies indicated that amylin contributes to the anorectic effects of cholecystokinin (CCK) and bombesin (BBS), possibly by enhancing the release of pancreatic amylin or by modulating their anorectic actions within the central nervous system (CNS). To elucidate the interaction between amylin and CCK or BBS, respectively, we investigated the influence of an IP injection of CCK or BBS on feeding in amylin-deficient mice (IAPP(-/-)). The anorectic effects of CCK and BBS were nearly abolished in IAPP(-/-) mice compared to wildtype (WT) mice (e.g. 20 microg/kg CCK, 1-h food intake: WT/NaCl 0.53 +/- 0.03 g; WT/CCK 0.16 +/- 0.03 g (P < 0.001); IAPP(-/-)/NaCl 0.49 +/- 0.05 g; IAPP(-/-)/CCK 0.39 +/- 0.04 g). Acute amylin replacement restored the anorectic effect of CCK in IAPP(-/-) mice. To find out whether CCK or BBS enhance the feeding-induced release of pancreatic amylin, we injected rats with CCK-8 (0.5-50 microg/kg) or BBS (5 microg/kg) and measured plasma amylin levels after injections. Neither CCK nor BBS increased the plasma amylin level in rats. We suggest that the mediation of the anorectic effects of CCK and BBS by amylin is not dependent on a CCK- or BBS-induced release of pancreatic amylin, but may rather be due to a modulation of their effects by amylin within the CNS. PMID- 12576090 TI - Impaired angiogenesis in neuropeptide Y (NPY)-Y2 receptor knockout mice. AB - Which of Y1-Y5 receptors (Rs) mediate NPY's angiogenic activity was studied using Y2R-null mice and R-specific antagonists. In Y2R-null mice, NPY-induced aortic sprouting and in vivo Matrigel capillary formation were decreased by 50%; Y1R antagonist blocked the remaining response. NPY-induced sprouting was equally inhibited by Y2R- (and Y5R- but less by Y1R-) antagonists in wild type mice. Spontaneous and NPY-induced revascularization of ischemic gastrocnemius muscles were similarly reduced in Y2R-null mice. Thus, NPY-induced angiogenesis, spontaneous and ischemic, is primarily mediated by Y2Rs. However, Y5Rs and, to a lesser degree Y1Rs, also may play a role in NPY-mediated angiogenesis. PMID- 12576091 TI - Synthesis and angiogenetic activity in the chick chorioallantoic membrane model of thymosin beta-15. AB - Thymosin beta-15 (Tbeta15), a 44 amino acid peptide (MW = 5173) localized in human prostate and breast cancer tissues was successfully synthesized in multigram quantities using Fmoc solid-phase peptide synthesis. The synthesized product was shown to have the right structure by ESI and MALDI mass spectral techniques and amino acid analysis. Relatively high yield was achieved, which might be due to enhanced acid stability of the p-cyanotrityl resin used. The effect of the synthesized Tbeta15 on the angiogenesis process was investigated using the chick chorioallantoic membrane (CAM) in vivo model. At concentrations above 1 microg/10 microl per disc, Tbeta15 exhibited a positive effect on angiogenesis, comparable to the effect of the intense angiogenetic factor phorbol 12-myristate 13-acetate at a standard concentration of 0.1 microg/10 microl per disc. The results of this study contribute to the further elucidation of the biological regulatory role of thymosin peptides and provide helpful information in the investigation of their possible therapeutic potential. PMID- 12576092 TI - Effects of adrenomedullin on the contraction of gastric arteries during reserpine induced gastric ulcer. AB - Adrenomedullin (100 ng/kg, s.c.) prevents reserpine-induced damage of gastric mucosa. In the model of in vitro gastric arteries from reserpine-treated rats, adrenomedullin pre-treatment resulted in a decrease of the vasoconstriction in response to 5-hydroxytryptamine. In contrast, adrenomedullin pre-treatment of rat with intact gastric mucosa did not affect the vasoconstriction to 5 hydroxytryptamine. In the presence of the NOS inhibitor N(G)-nitro-L-arginine, the responsiveness to 5-hydroxytryptamine in gastric arteries from rats treated with reserpine + adrenomedullin was enhanced to the same level of rats treated with reserpine alone. The anti-ulcer effect of adrenomedullin could therefore be related, at least in part, to an increase of blood flow at the gastric mucosa, by a mechanism involving nitric oxide. PMID- 12576093 TI - Octreotide improves burn-induced intestinal injury in the rat. AB - The local thermal trauma activates a number of systemic mediator cascades, e.g. a complement activation, cytokine production, resulting in a generalized sequestration and a priming of local and systemic neutrophils and macrophages. We aimed to determine the possible protective effect of octreotide (OCT), a synthetic somatostatin analogue, against burn-induced intestinal tissue damage possibly by inhibiting neutrophil infiltration. Under brief ether anaesthesia, shaved dorsum of the rats was exposed to 90 degrees C bath for 10s to induce burn injury. Rats were decapitated either 3, 24 or 72 h after burn injury. Octreotide (10 microg/kg) or saline was administered subcutaneously (s.c.) immediately after the burn injury. In the 24- and 72-h burn groups, OCT injections were repeated three times daily. In the sham group the same protocol was applied except that the dorsum was dipped in a 25 degrees C water bath for 10 s Malondialdehyde (MDA) and glutathione (GSH) levels and myeloperoxidase (MPO) activity were determined in the intestinal tissue. The results demonstrate that burn injury results in significant neutrophil accumulation, as evidenced by increases in MPO activity. The increase in MDA and the concomitant decrease in GSH levels demonstrate the role of oxidative mechanisms in burn injury. OCT may have some beneficial therapeutic effects by reducing neutrophil-dependent injury and related lipid peroxidation following burn trauma. PMID- 12576094 TI - Pinealectomy blocks modulation of active avoidance by central vasopressin application in rats. AB - The inter-relationship between central vasopressin and the pineal gland in the modulation of active avoidance behavior was investigated. In sham-operated (SO) rats, intracerebroventricular (i.c.v) application of 10 ng arginine vasopressin (AVP) after both the last acquisition and the first extinction trials prolonged the extinction of the active avoidance response; application of 50 ng of the V1 antagonist, d(CH2)5Tyr(Me)AVP (AAVP) was without effect in both experiments. In contrast to the SO in pinealectomized (PX) rats neither AVP nor AAVP influenced the extinction of the avoidance response. Intraseptal infusion of 200 pg AVP or 5 ng AAVP either after the last acquisition or the first extinction trial was without effect in both SO and PX rats. Comparison of the acquisition trials revealed no differences between SO and PX rats. PMID- 12576095 TI - PLG regulates hnRNP-L expression in the rat striatum and pre-frontal cortex: identification by ddPCR. AB - Central dopaminergic systems are implicated in schizophrenia and Parkinson's disease, and are known to be modulated by the endogenous tripeptide Pro-Leu-Gly NH(2) (PLG or MIF-1, melanocyte-stimulating hormone release inhibiting factor-1). Differential display polymerase chain reaction (ddPCR) was utilized to identify genes that are regulated by protracted PLG treatment (20 mg/kg, i.p. for 28 days) in male Sprague-Dawley rats. A total of 2400 genes were screened and 3 down regulated bands were identified in the PLG-treated samples. Sequencing analysis revealed a total of six unique cDNA species. One fragment possessed a high degree of homology with Mus musculus hnRNP-L (protein L) mRNA (GenBank #AB009392) (termed PRG1: PLG regulated gene 1). Elongation of the PRG1 cDNA, by RACE-PCR, provided an 835 bp sequence with 95% homology to AB009392 over a 743 bp span. Open reading frame analysis provided a putative amino acid sequence consistent with the identity of PRG1 as rat hnRNP-L. Northern hybridization experiments with PRG1 revealed a 2.3 kb mRNA species that was decreased by 65% in the PLG-treated tissue. Western blot analysis revealed significantly decreased hnRNP-L levels in the striatum and pre-frontal cortex (but not the nucleus accumbens) by 71 and 61%, respectively of PLG-treated animals. The identification of altered expression of hnRNP-L following PLG treatment provides insight into the long-term effects of PLG and may provide insight into its molecular mechanism of action. PMID- 12576096 TI - Substance P(1-7) affects the expression of dopamine D2 receptor mRNA in male rat brain during morphine withdrawal. AB - Previous studies have confirmed an important role of the undecapeptide substance P (SP) in opioid reward and dependence. It is further shown that the SP N terminal metabolite SP(1-7) may attenuate the intensity of opioid withdrawal in mice. In this study we have investigated the effect of the heptapeptide fragment on the expression of the brain dopamine D2 receptor mRNA and on the withdrawal reaction, as well, in morphine-dependent rats. Male Sprague-Dawley rats were randomly distributed into two groups. Guide cannula was implanted and aimed at the lateral ventricle and animals were subsequently made opioid dependent by two daily injections of morphine (10 mg/kg) for 7 days. Half an hour before naloxone challenge (2 mg/kg) one group of rats received an injection of SP(1-7) (28 nmol per rat) and the other, serving as control, was injected with saline through the cannula. Animals were decapitated 4 h following SP(1-7) or saline injections. The results indicated that the level of the dopamine D2 receptor transcript was significantly reduced by SP(1-7) in nucleus accumbens and frontal cortex but not altered in the striatum. In behavioral tests it was found that the heptapeptide attenuated several somatic withdrawal symptoms. The observed reduction in the receptor transcript in nucleus accumbens and frontal cortex is suggested to reflect an increased dopamine activity in these areas, which in turn may counteract the withdrawal reaction. PMID- 12576097 TI - Histamine H1 receptors in the ventromedial hypothalamus mediate the anorectic action of the pancreatic hormone amylin. AB - In the present study we investigated the role of hypothalamic histamine H(1) receptors in the mediation of peripheral amylin's anorectic effect. Rats with chronically implanted bilateral cannulas were infused into the ventromedial hypothalamus (VMH) with the specific histamine H(1) receptor antagonists pyrilamine (PYR, 104 nmol/rat) or chlorpheniramine (CPA, 52 nmol/rat), respectively, combined with an intraperitoneal (IP) injection of amylin (5 microg/kg). Amylin's inhibitory effect on food intake (i.e. 50% reduction in cumulative food intake 30 min after ingestion) was markedly reduced by CPA and PYR (e.g. amylin and CPA: 5% reduction versus control). We therefore suggest an important role of hypothalamic H(1) receptors in the signal transduction of peripheral amylin's anorectic action. PMID- 12576098 TI - Support vector machines for prediction of protein signal sequences and their cleavage sites. AB - Given a nascent protein sequence, how can one predict its signal peptide or "Zipcode" sequence? This is an important problem for scientists to use signal peptides as a vehicle to find new drugs or to reprogram cells for gene therapy (see, e.g. K.C. Chou, Current Protein and Peptide Science 2002;3:615-22). In this paper, support vector machines (SVMs), a new machine learning method, is applied to approach this problem. The overall rate of correct prediction for 1939 secretary proteins and 1440 nonsecretary proteins was over 91%. It has not escaped our attention that the new method may also serve as a useful tool for further investigating many unclear details regarding the molecular mechanism of the ZIP code protein-sorting system in cells. PMID- 12576099 TI - VIP as a trophic factor in the CNS and cancer cells. AB - The effects of vasoactive intestinal peptide (VIP) on the proliferation of central nervous system (CNS) and cancer cells were investigated. VIP has important actions during CNS development. During neurogenesis, VIP stimulates the proliferation and differentiation of brain neurons. Addition of VIP to embryonic mouse spinal cord cultures increases neuronal survival and activity dependent neurotrophic factor (ADNF) secretion from astroglial cells. VIP is an integrative regulator of brain growth and development during neurogenesis and embryogenesis. Also, VIP causes increased proliferation of human breast and lung cancer cells in vitro. VIP binds with high affinity to cancer cells, elevates the cAMP and increases gene expression of c-fos, c-jun, c-myc and vascular endothelial cell growth factor. The effects of VIP on cancer cells are reversed by VIPhybrid, a synthetic VPAC(1) receptor antagonist. VIPhyb inhibits the basal growth of lung cancer cells in vitro and tumors in vivo and potentiates the ability of chemotherapeutic drugs to kill cancer cells. Due to the high density of VPAC(1) receptors in cancer cells, VIP has been radiolabeled with 123I, 18F and 99mTc to image tumors. It remains to be determined if radiolabeled VIP analogs will be useful agents for early detection of cancer in patients. PMID- 12576101 TI - Meta-learning in reinforcement learning. AB - Meta-parameters in reinforcement learning should be tuned to the environmental dynamics and the animal performance. Here, we propose a biologically plausible meta-reinforcement learning algorithm for tuning these meta-parameters in a dynamic, adaptive manner. We tested our algorithm in both a simulation of a Markov decision task and in a non-linear control task. Our results show that the algorithm robustly finds appropriate meta-parameter values, and controls the meta parameter time course, in both static and dynamic environments. We suggest that the phasic and tonic components of dopamine neuron firing can encode the signal required for meta-learning of reinforcement learning. PMID- 12576102 TI - Learning to generate articulated behavior through the bottom-up and the top-down interaction processes. AB - A novel hierarchical neural network architecture for sensory-motor learning and behavior generation is proposed. Two levels of forward model neural networks are operated on different time scales while parametric interactions are allowed between the two network levels in the bottom-up and top-down directions. The models are examined through experiments of behavior learning and generation using a real robot arm equipped with a vision system. The results of the learning experiments showed that the behavioral patterns are learned by self-organizing the behavioral primitives in the lower level and combining the primitives sequentially in the higher level. The results contrast with prior work by Pawelzik et al. [Neural Comput. 8 (1996) 340], Tani and Nolfi [From animals to animats, 1998], and Wolpert and Kawato [Neural Networks 11 (1998) 1317] in that the primitives are represented in a distributed manner in the network in the present scheme whereas, in the prior work, the primitives were localized in specific modules in the network. Further experiments of on-line planning showed that the behavior could be generated robustly against a background of real world noise while the behavior plans could be modified flexibly in response to changes in the environment. It is concluded that the interaction between the bottom-up process of recalling the past and the top-down process of predicting the future enables both robust and flexible situated behavior. PMID- 12576103 TI - Computational model for neural representation of multiple disparities. AB - It has been known that the visual system can detect more than one disparity and/or motion direction at the same region in the image. These multiple (or transparent) surfaces can be perceived when, for example, we are looking scenes through a glass. However, many conventional models cannot deal with these multiple surfaces. The present paper investigates the neural encoding and decoding of multiple disparities with the binocular energy model, which is known as a biologically plausible model. Based on the analysis of the response of the energy model to multiple disparities, the present paper proposes a stereo model that can detect disparities of two overlapping surfaces. PMID- 12576104 TI - A synfire chain in layered coincidence detectors with random synaptic delays. AB - In this paper we analyze a synfire chain in a spiking neuron network. We also employ a coincidence-detector model showing the characteristics of temporal information processing more directly than the integrate-and-fire (I&F) model often discussed in the literature. There are two sources of randomness in a feed forward network, however, only randomness in input spikes has attracted the attention of researchers and the randomness in synaptic delays has largely been ignored. Theoretical analyses of the synfire chain in I&F neurons without randomness in synaptic delays have shown that the dynamics of pulse packets can be viewed as a shift of the membrane potential distribution made by random noise input spikes. We introduce jittered synaptic delays instead of random noise inputs in a network of coincidence detectors and show that the network has almost the same dynamics as that of the I&F neurons. The distribution of the output spikes can be approximately described by an ordinary differential equation useful in understanding the dynamics of the pulse packets. PMID- 12576105 TI - Mathematical analysis of a correlation-based model for orientation map formation. AB - We consider a correlation-based model for the orientation map formation proposed by Miller [Journal of Neuroscience 14 (1994) 409] and study the formation mathematically. We perform the Fourier transform and compute the principal component of the model. With our analysis, the roles of functions are clarified and the result indicates that the developed orientation maps have the following properties. (1) Maps have oriented receptive fields. (2) Preferred orientations smoothly change on the cortical surface. (3) Periodicity of preferred orientations does not appear. (4) Periodicity of phases appears. (5) Singular points appear irregularly on the cortical surface. Our analytical results are justified by computer simulations. PMID- 12576106 TI - The functional localization of neural networks using genetic algorithms. AB - We presented an algorithm for extracting Boolean functions (propositions, rules) from the units in trained neural networks. The extracted Boolean functions make the hidden units understandable. However, in some cases, the extracted Boolean functions are complicated, and so are not understandable, which means that the hidden units are not functionally localized. This paper presents an algorithm for the functional localization of (the hidden units of) neural networks. When a hidden unit is well approximated to a low-order Boolean function, the unit can be regarded as functionally localized. The functional localization of a hidden unit is evaluated by the error between the hidden unit and the low-order Boolean function extracted from the hidden unit. The optimization is executed by genetic algorithms. We applied it to vote data, mushroom data and chess data. Experimental results show that the algorithm works well. PMID- 12576107 TI - A new EM-based training algorithm for RBF networks. AB - In this paper, we propose a new Expectation-Maximization (EM) algorithm which speeds up the training of feedforward networks with local activation functions such as the Radial Basis Function (RBF) network. In previously proposed approaches, at each E-step the residual is decomposed equally among the units or proportionally to the weights of the output layer. However, these approaches tend to slow down the training of networks with local activation units. To overcome this drawback in this paper we use a new E-step which applies a soft decomposition of the residual among the units. In particular, the decoupling variables are estimated as the posterior probability of a component given an input-output pattern. This adaptive decomposition takes into account the local nature of the activation function and, by allowing the RBF units to focus on different subregions of the input space, the convergence is improved. The proposed EM training algorithm has been applied to the nonlinear modeling of a MESFET transistor. PMID- 12576108 TI - Analysis of Tikhonov regularization for function approximation by neural networks. AB - This paper is devoted to the convergence and stability analysis of Tikhonov regularization for function approximation by a class of feed-forward neural networks with one hidden layer and linear output layer. We investigate two frequently used approaches, namely regularization by output smoothing and regularization by weight decay, as well as a combination of both methods to combine their advantages. We show that in all cases stable approximations are obtained converging to the approximated function in a desired Sobolev space as the noise in the data tends to zero (in the weaker L(2)-norm) if the regularization parameter and the number of units in the network are chosen appropriately. Under additional smoothness assumptions we are able to show convergence rates results in terms of the noise level and the number of units in the network. In addition, we show how the theoretical results can be applied to the important classes of perceptrons with one hidden layer and to translation networks. Finally, the performance of the different approaches is compared in some numerical examples. PMID- 12576109 TI - Predicting the behaviour of G-RAM networks. AB - A Generalising Random Access Memory (G-RAM) neuron is distinguished from conventional neuron models by the fact that its tolerance to departures in similarity from its training pattern is variable. Details of this are given in this paper as it affects the behaviour a class of digital probabilistic neural networks which have been achieving attention in the neural networks literature for some years now. Such systems are also called n-tuple systems, weightless systems or p-RAM systems. After reviewing the literature on such networks, a novel simple combinatoric analysis of the most likely behaviour of recursive GRAM networks is described. The best network performance, measured by a key parameter called 'radius of retrievability' (first defined by Wong and Sherrington [J. Phys. A 22 (1989) 2233] as the error in the input that still allows evolution of the dynamic network to the correct attractor state), is obtained with a training set composed of random data patterns. Increasing the size of the training set reduces this radius of retrievability in a predictable manner. Changing the nature of the training set to non-random patterns also reduces the radius of retrievability to an extent that we show can be estimated from a measure of the diversity of the elements of the training set (we refer to this as the 'mean intra-set Hamming distance of the training set'). As mentioned earlier the feature of G-RAMs (indicated by the G) is that there exists a generalization parameter which determines how far a neuron input vector can stray from a training input for the neuron to respond in the trained way. It is shown that when this generalization parameter is reduced, then the radius of retrievability is also reduced but it is then stable in the face of an increase in size, or change in nature, of the training set. This is a novel prediction of the behaviour of systems and of the robustness of such behaviour in the face of varying the size and correlation properties of the training set. PMID- 12576110 TI - On-line identification and reconstruction of finite automata with generalized recurrent neural networks. AB - In this paper finite automata are treated as general discrete dynamical systems from the viewpoint of systems theory. The unconditional on-line identification of an unknown finite automaton is the problem considered. A generalized architecture of recurrent neural networks with a corresponding on-line learning scheme is proposed as a solution to the problem. An on-line rule-extraction algorithm is further introduced. The architecture presented, the on-line learning scheme and the on-line rule-extraction method are tested on different, strongly connected automata, ranging from a very simple example with two states only to a more interesting and complex one with 64 states; the results of both training and extraction processes are very promising. PMID- 12576111 TI - Classification of clustered microcalcifications using a Shape Cognitron neural network. AB - A new shape recognition-based neural network built with universal feature planes, called Shape Cognitron (S-Cognitron) is introduced to classify clustered microcalcifications. The architecture of S-Cognitron consists of two modules and an extra layer, called 3D figure layer lies in between. The first module contains a shape orientation layer, built with 20 cell planes of low level universal shape features to convert first-order shape orientations into numeric values, and a complex layer, to extract second-order shape features. The 3D figure layer is a feature extract-display layer that extracts the shape curvatures of an input pattern and displays them as a 3D figure. It is then followed by a second module made up of a feature formation layer and a probabilistic neural network-based classification layer. The system is evaluated by using Nijmegen mammogram database and experimental results show that sensitivity and specificity can reach 86.1 and 74.1%, respectively. PMID- 12576112 TI - On learning to estimate the block directional image of a fingerprint using a hierarchical neural network. AB - This paper presents a hierarchical neural network architecture for computing fingerprints block directional images. Two separately trained neural networks are connected in series. First, the fingerprint image is divided into 16x16 blocks, each block is submitted to the first network which is a back propagation neural network. It has four counters in its output layer one for each direction to count the main directional codes in each fingerprint block. The output of this network is considered the feature vector for the fingerprint block, which is then submitted to the second network. The second network is a self-organized feature maps neural network uses an unsupervised learning strategy to group the fingerprint blocks into distinct directional classes. In this scheme, there is more than one sub-class for each directional class, an agglomerative hierarchical cluster algorithm for merging two clusters is used to merge two classes if their corresponding distances are below a specified threshold. Results obtained with a real world data set indicate the effectiveness of the proposed architecture. PMID- 12576114 TI - Conditioned acceptance and preference but not altered taste reactivity responses to bitter and sour flavors paired with intragastric glucose infusion. AB - Nutrient-conditioned flavors preferences are thought to involve an increase in flavor palatability (hedonic evaluation). Consistent with this view is the recent finding that a sweet flavor paired with intragastric glucose infusions elicited more hedonic taste reactivity (TR) responses than did an alternative sweet flavor paired with intragastric water. The generality of this finding was examined by conditioning preferences for inherently avoided nonsweet flavors. Rats were trained in 20 h/day and then 30 min/day sessions with a CS+ flavor (sour citric acid or bitter sucrose octaacetate) paired with intragastric 16% glucose infusion, and the opposite flavor (CS-) paired with intragastric water. Glucose conditioning increased the CS+ acceptance in one-bottle tests and produced a 95% CS+ preference in two-bottle sessions. Yet, TR responses to brief intraoral infusions of the two CS flavors did not differ, even after extensive testing. Subsequent choice tests revealed that a 1% fructose solution was preferred to the CS-, whereas the CS+ was preferred to 1% and 2% fructose and equally preferred to 4%, 8%, and 16% fructose. These results indicate that strong nutrient-conditioned flavor preferences are not always associated with increased flavor palatability as measured by TR tests. Therefore, nonhedonic processes, perhaps increased incentive salience, appear to mediate the enhanced preference and acceptance conditioned by postingestive nutrient actions. PMID- 12576115 TI - Effects of alcohol exposure during development on social behavior in rats. AB - In addition to the cognitive deficits associated with fetal alcohol syndrome (FAS), clinical and animal studies indicate that alcohol exposure might also have detrimental effects on social behavior. In a rat model of FAS, experimental rats were given alcohol from gestational day (GD) 1 to 22 and from postnatal day (PD) 2 to 10, a period roughly equivalent to all three trimesters in humans. Control groups consisted of rats exposed to the administration procedures but not to alcohol and nontreated rats. At 30 days of age, rats were tested for social behavior in an alley maze that contained its cagemate in the goal box. After varying periods of isolation, the animals' latencies to reach the goal box and their social behaviors once inside the goal box were recorded. Alcohol-exposed animals ran faster than control rats to the occupied goal box regardless of the amount of isolation. The alcohol-exposed animals also exhibited aberrant social interactions with their cagemate once inside the goal box compared to one or both of the control groups. Specifically, the alcohol-exposed animals showed greater amounts of anogenital sniffing, chasing, hopping and darting, and retrieving and lesser amounts of pinning and biting compared to one or both of the control groups. The alcohol-induced change in anogenital sniffing varied over increasing amounts of isolation compared to both control groups, but the alterations in the other behaviors did not. It is argued that the altered social behavior of alcohol exposed animals is not the result of changes in the animals' motivational state or social learning and may be the result of an increased responsiveness to social stimuli. PMID- 12576116 TI - Assessment of the Sniff Magnitude Test as a clinical test of olfactory function. AB - Olfactory threshold and odor identification tasks currently serve as the standard approaches to the clinical assessment of olfactory function. Although these methods can be used effectively with the average patient, they suffer from some limitations when used to evaluate children, people with cognitive impairment, or people from diverse cultural backgrounds. A novel approach to the clinical evaluation of olfactory function, the Sniff Magnitude Test, attempts to minimize the cognitive demands of an olfactory test and thereby overcome some of the limitations of alternative tests. This is achieved by measuring the reflex-like reduction in sniffing that occurs when a malodor is encountered. The reliability and the validity of the Sniff Magnitude Test were assessed by testing people on two occasions using the University of Pennsylvania Smell Identification Test (UPSIT), a butanol threshold task, the Alcohol Sniff Test, and the Sniff Magnitude Test. The test-retest reliability of the Sniff Magnitude Test was r=.80, higher than the butanol thresholds and Alcohol Sniff Test but somewhat lower than the UPSIT. Performance on the Sniff Magnitude Test (the sniff magnitude to a malodor relative to nonodorized air) was correlated between r=-.61 and r=-.66 with the other measures of olfactory function. This range of correlations was comparable to that observed between the butanol threshold, the UPSIT, and the Alcohol Sniff Test. Finally, evidence for the advantages of the Sniff Magnitude Test, as compared to the UPSIT, was provided by a study with young children. It is concluded that the Sniff Magnitude Test has significant potential as a clinical measure of olfactory function, and that further testing and development of this method are warranted. PMID- 12576117 TI - Photoperiod and stress affect wound healing in Siberian hamsters. AB - Changes in day length alter several indices of immune function in Siberian hamsters. These experiments tested the hypothesis that photoperiodic changes in immune function are integrated at an organismal level as reflected by the ability to heal a cutaneous wound. Given the well-documented effects of psychological stressors on immune function, we also tested the hypothesis that photoperiod modulates the effects of acute stress on wound healing. Male hamsters were housed in long (16L:8D; LD) or short (8L:16D; SD) day lengths for 8+/-2 weeks. SD treatment was sufficient to induce winter reproductive status. Hamsters then received a dermal punch wound. Hamsters were subjected to either 2 h of daily restraint stress or a control treatment for 3 days prior to and 5 days after wounding. Wounds were digitally photographed daily, and wound size was measured to quantify healing. Wounds of LD hamsters healed significantly faster than did those of SD hamsters. Restraint stress significantly accelerated healing in SD hamsters. The results suggest that the enhancing effects of short-term psychological stressors on immune function are apparent only when reproductive function is suppressed. In nature, enhanced wound healing coincident with the breeding season and territorial defense may be adaptive. PMID- 12576118 TI - Effects of an odor paired with illness on startle, freezing, and analgesia in rats. AB - The data reported in this experiment provide the first systematic exploration of the effectiveness of an odor previously paired with an aversive reinforcer other than shock on eliciting various behavioral expressions of fear in the rat. Specifically, we measured potentiation of the acoustic startle response, freezing, and analgesia in the presence of an odor previously paired with an illness-inducing agent (lithium chloride; LiCl). We found that this odor elicited freezing and analgesia, but failed to potentiate the startle response. The results are discussed in terms of (1). potential threshold differences for various expressions of learned fear and (2). the possibility that the content of the learning established by odor-shock pairings differ from those established by odor-illness pairings. PMID- 12576119 TI - Effects of deprivation on hedonics and reinforcing value of food. AB - Eating is influenced by both the hedonic preferences and reinforcing value of food. Incentive salience theory predicts these are separate influences. This study tested whether hedonics reliably change as a function of increasing the reinforcing value of food by deprivation in 17 non-obese, non-dietary restrained females. Baseline measures of hedonics for pleasant (chocolate milk), unpleasant (lemon juice) and neutral (water) tastes were determined under deprived conditions. Participants were then randomly assigned to fed or maintained in deprived conditions, and after food consumption, a second determination of hedonics was obtained, followed by assessing the reinforcing value of food. Hedonics was measured by subjective ratings and behavioral observations in a taste reactivity paradigm. Results showed food was more reinforcing for the deprived than the fed group, but no influences of group were observed for the subjective or objective hedonic measures. These results suggest that hedonics and the reinforcing value of food are separate processes in humans, and they may independently influence eating behavior. PMID- 12576120 TI - Effect of the serotonin agonist buspirone on behaviour and hypothalamic-pituitary adrenal axis in confident and fearful mink. AB - Behavioural and hypothalamic-pituitary-adrenal (HPA) axis responses were investigated in farm mink (Mustela vison) selected for either confident or fearful behaviour for nine generations. Two groups of 2-year-old confident (n=12) and fearful (n=12) female mink were given the serotonin (5-HT) 1A receptor agonist buspirone (1.25 mg/kg/day), whereas two other groups of 2-year-old confident (n=12) and fearful (n=12) female mink were given saline, continuously for 5 weeks via osmotic minipumps. Behavioural reactions towards a novel object and towards humans were tested after 19-25 days, and HPA axis reactivity [adrenocorticotropic hormone (ACTH), cortisol] was measured after 28-31 days of treatment. Confident mink were more exploratory than fearful mink towards humans and a novel object. Confident mink spent more time in contact with the object than did fearful mink during saline-but not during buspirone-treatment. buspirone increased approach-withdrawal conflict behaviour towards a object in fearful mink only. The chronic dose of buspirone did not reduce fear towards humans and did not affect latencies to reaction, number of contacts, number and duration of manipulations, and stereotypic behaviour in a Novel Object test. Different HPA axis responses have emerged between confident and fearful mink, together with a different degree of fear-related behaviour. Fearful mink have a higher cortisol combined with a lower ACTH secretion than confident mink in response to capture and blood sampling. The central serotonergic system may be involved, and even though the precise underlying mechanisms are presently unknown, treatment with a 5-HT(1A) receptor agonist reduces the difference between confident and fearful mink in HPA axis reactivity. PMID- 12576121 TI - Sleep and EEG patterns in the chronic constriction injury model of neuropathic pain. AB - Chronic neuropathic pain patients often report sleep disturbances such as reduced amount of sleep and excessive daytime tiredness. The aim of this study was to evaluate possible abnormalities in sleep patterns in a widely used animal model of neuropathic pain. Adult male Sprague-Dawley rats were chronically implanted with electrodes for electroencephalogram (EEG) and electromyogram (EMG) registrations to allow continuous 24-h polygraphic recording. Subsequently, a chronic constriction injury (CCI) was inflicted on eight rats in accordance with the CCI model of neuropathic pain and a sham operation was performed on another eight rats. The polygraphic recordings were repeated 13, 27, 55, and 146 days after surgery. Although the CCI animals developed significant mechanical and cold allodynia and heat hyperalgesia, there were no significant differences between the CCI rats and the sham-operated control animals in the spontaneous EEG/EMG in homecage-like conditions. It is concluded that in the chronic phase, this neuropathic pain model does not produce clear sleep disturbances. Such an absence of general suffering from sleep disturbances is advantageous to the CCI model as it makes use of the model more acceptable ethically. Nonetheless, this outcome appears to be in contrast with the clinical situation in neuropathic pain and therefore could also be seen as a disadvantage for the face validity of the CCI model. PMID- 12576122 TI - Effects of fat content on fat hedonics: cognition or taste? AB - Understanding and perhaps overriding preferences for fat is important, given the relationship between higher dietary fat consumption and poorer health. We have examined the roles of potential mechanisms for differences in fat preference: actual fat content and expected fat content. The subjects were women (n=192, ages=50-69) recruited to a study of low-fat dietary change. Subjects were randomized to one of the four cells: participants received either a high- or low fat milkshake at baseline, and half of each group was told that their milkshake was low in fat and the other half high in fat. Women who received a high-fat milkshake consumed more grams than women who received a low-fat milkshake. Women who expected low-fat shakes reported liking them more than those who expected high-fat milkshakes. These data indicate that both physiology and cognition play a role in determining consumption of high- and low-fat foods. PMID- 12576123 TI - Cocaine-induced genital reflexes during paradoxical sleep deprivation and recovery. AB - Paradoxical sleep deprivation (PSD) for 96 h together with cocaine administration elicits genital reflexes (penile erection [PE] and ejaculation [EJ]) in rats. Our objective was to examine genital reflexes after periods of 24, 48, 72, 96, 120, and 144 h of PSD and during a 4-day recovery period in acute cocaine-administered rats. After 24 h of PSD followed by cocaine administration, animals started to display PE and EJ, peaking in the 96th h of PSD, whereas PE and EJ were absent in control animals. The effects of more than 96 h of PSD decrease genital reflexes as observed after 120 and 144 h. Genital reflexes were present in the recovery periods but diminished gradually during the period evaluated. Even short periods of PSD probably cause supersensitivity of dopamine (DA) receptors and exacerbate the effects of cocaine on dopaminergic pathways to induce frequent PE and EJ. PMID- 12576124 TI - A sedentary day: effects on subsequent sleep and body temperatures in trained athletes. AB - Exercise effects on sleep in fit healthy people have been difficult to determine because their sleep is close to optimal, leaving little room for improvement. Another method for assessing exercise effects on sleep is to significantly reduce the degree of activity in highly active people. Fifteen trained athletes who exercised daily at a moderate to high intensity were employed. By requesting that subjects remain sedentary in the laboratory for an entire day, the effect of reduced exercise on subsequent sleep parameters was assessed. Sleep and temperature were recorded after a sedentary day and after a normal day of moderate to high activity (control condition) in a counterbalanced design. In the sedentary condition, slow-wave sleep (SWS) decreased by a mean of 15.5+/-7.0 min and slow-wave activity (SWA) differed significantly (P<.05) between conditions in the first hour of sleep only. Rapid eye movement (REM) sleep increased by a mean of 17.9+/-5.7 min in the sedentary condition, while sleep onset latency (SOL) to Stages 1 and 2 increased by 10.2 and 10.7 min, respectively, and REM sleep latency decreased by 24.0+/-6.8 min (all P<.05). Between conditions, there was no overall effect on total sleep time (TST), sleep efficiency, wake after sleep onset or core or foot temperatures (P>.05). With reduced exercise load, SWS pressure may have been reduced, resulting in lower levels of SWS and increased REM sleep. Thus, the data indicate that reducing exercise has significant effects on sleep that may have implications for athletes tapering for competition. PMID- 12576125 TI - Testosterone, territoriality, and the 'home advantage'. AB - The consistently better performance seen by teams in various sporting contexts when playing at home is referred to as the 'home advantage'. Various explanations have been put forward to account for this robust phenomenon, though none has yet focussed on possible hormonal factors. In an initial study, we showed that salivary testosterone levels in soccer players were significantly higher before a home game than an away game.In a second study involving a different group of soccer players, this finding was replicated over two home games, two away games, and three training sessions. Perceived rivalry of the opposing team was important as testosterone levels were higher before playing an 'extreme' rival than a 'moderate' rival. Self-reported measures of mood in both studies were not linked to testosterone level. The present results corroborate and extend earlier findings on the relationships between testosterone, territoriality, and dominance in human competitive encounters and further suggest an important role for testosterone in the home advantage seen in various team sports. PMID- 12576126 TI - Exogenous adrenocorticotrophic hormone does not elicit a salt appetite in growing pigs. AB - In rodents, rabbits, and sheep, exogenous adrenocorticotrophic hormone (ACTH) leads to a marked increase in sodium appetite. It has been suggested that if pigs show a similar response to stress, an appetite for salt could increase their attraction to blood and contribute to the development of tail biting. The aim of this study was to examine the effects of ACTH on salt appetite in growing pigs. Individually housed Yorkshire pigs (45 kg) were divided into three groups of four. Group 1 had free access to water, 0.5 M NaCl, and 0.5 M KCl solutions; Group 2 to water, 0.5, and 0.25 M NaCl solutions; Group 3 to water, 0.25, and 0.125 M NaCl solutions. Intramuscular injection of long-acting synthetic ACTH (50 IU twice daily for 5 days) did not elicit increases in intakes of any of the available salt solutions compared to pretreatment intakes. However, there was a 1.6-fold increase in both water and feed intake during ACTH treatment. ACTH treatment also stimulated significant increases in salivary cortisol concentrations. Although increases in salivary cortisol concentrations and in water and feed intake indicate that there were physiological responses to the treatment, exogenous ACTH given for 5 days did not elicit a sodium appetite in growing pigs. These findings do not support the notion that a stress-induced salt appetite serves as an underlying mechanism for tail biting. PMID- 12576127 TI - Selective effects of vagal deafferentation and celiac-superior mesenteric ganglionectomy on the reinforcing and satiating action of intestinal nutrients. AB - The role of vagal afferents and splanchnic fibers in nutrient-induced flavor conditioning and feeding suppression was determined. Male rats were fitted with intraduodenal (ID) catheters and given subdiaphragmatic vagal deafferentation (SDA), celiac-superior mesenteric ganglionectomy (CGX), combined (COM) treatments, or sham surgery. In separate conditioning trials, they were trained to drink (30 min/day) flavored saccharin solutions paired with concurrent ID infusions of 8% maltodextrin or water and 3.55% corn oil or water. Experiment 1 revealed that SDA and sham rats showed equal preferences for the nutrient-paired flavors over the water-paired flavors. In contrast, SDA rats, unlike sham rats, failed to suppress their intake of a palatable fluid when infused intraduodenally with maltodextrin or corn oil. Experiment 2 revealed that CGX, COM and sham rats all developed preferences for the maltodextrin-paired flavor, although CGX alone or COM attenuated the conditioned preference. CGX and COM treatments also attenuated or blocked the feeding inhibitory actions of ID nutrient infusions. These findings along with prior data indicate that gut vagal afferents and splanchnic nerves are not essential for flavor-nutrient preference conditioning, whereas both vagal afferents and splanchnic nerves are implicated in carbohydrate and fat-induced satiation. PMID- 12576128 TI - Effect of repeated administration of dexfenfluramine on feeding and brain Fos in mice. AB - The present study examines whether repeated administration of dexfenfluramine (DFEN) to mice is associated with progressive attenuation of its anorectic action and induction of Fos-immunoreactivity (Fos-ir) in some brain regions, both known effects in rats. ICR:Cd-1 mice were adapted to receiving for 30 min daily a sweetened milk gelatin dessert in addition to ad libitum access to chow. Mice were then injected with either DFEN (5-10 mg/kg) or saline vehicle every other day. These doses of DFEN acutely suppressed intake by approximately 50% and this action was sustained for at least eight injections. The anorectic effect of DFEN was no different between drug-naive and DFEN-pretreated mice under conditions that produce complete tolerance in rats. In addition, unlike in rats, the acute anorectic effect of m-chlorophenylpiperazine (mCPP) was unaffected by prior DFEN treatment. Mice were perfused after the intake test on the final experimental day and their brain processed for Fos-ir. DFEN induced Fos-ir in brain regions analogous to those reported in rats and, also, as in rats, DFEN pretreatments reduced substantially the Fos-ir induced by acute DFEN in regions including central nucleus of amygdala, bed nucleus of stria terminalis, and the paraventricular hypothalamus (magno- and parvocellular divisions; PVN). Thus, while DFEN pretreatments reduce the effects of DFEN in many brain regions in both rats and mice, mice are unlike rats insofar as there is no tolerance to the anorectic action, at least in the nondeprivation protocol used here. PMID- 12576129 TI - Stress-induced impairment of inhibitory avoidance learning in female neuromedin B receptor-deficient mice. AB - Neuromedin B (NMB) is a mammalian bombesin (BN)-like peptide that exerts its function via the neuromedin B receptor (NMB-R). The NMB/NMB-R system is involved in stress response, and therefore we examined behavioral properties in female mice lacking NMB-R using a restraint-induced stress paradigm. Thirty minutes of restraint in a wire mesh cage constituted a sufficient stress stimulus for mice as evidenced by elevated blood glucose concentrations in stressed wild-type and NMB-R-deficient mice. Using a one-trial passive avoidance test, stressed NMB-R deficient mice exhibited a marked reduction in memory performance. NMB-R deficient mice exhibited elevated spontaneous activity in a novel environment compared to non-stressed mutant mice after 30-min stress, and a similar difference was also observed between stressed/non-stressed wild-type mice. An elevated plus maze test showed that the stress stimulus had no effect on anxiety in either wild-type or NMB-R-deficient mice. Furthermore, pain response of wild type and NMB-R-deficient mice induced by electric foot shock was not affected under either stressed or non-stressed conditions. These results indicate that impaired memory performance in stressed NMB-R-deficient mice is not a consequence of changes in spontaneous activity, anxiety, or pain response, and suggest that the NMB/NMB-R pathway may play a role in regulating the stress response via the neural system that controls learning and memory. PMID- 12576130 TI - A high-protein diet enhances satiety without conditioned taste aversion in the rat. AB - In order to determine the respective roles of conditioned food aversion, satiety and palatability, we studied behavioral responses to a 50% total milk protein diet, compared with those to a normal protein diet containing 14% total milk protein. Different paradigms were employed, including meal pattern analysis, two choice testing, flavor testing, a behavioral satiety sequence (BSS) and taste reactivity. Our experiments showed that only behavioral and food intake parameters were disturbed during the first day when an animal ate the high protein (P50) diet, and that most parameters returned to baseline values as soon as the second day of P50. Rats adapted to P50 did not acquire a conditioned taste aversion (CTA) but exhibited satiety, and a normal BSS. The initial reduction in high-protein diet intake appeared to result from the lower palatability of the food combined with the satiety effect of the high-protein diet and the delay required for metabolic adaptation to the higher protein level. PMID- 12576131 TI - Successful acquisition of an olfactory discrimination paradigm by spider monkeys, Ateles geoffroyi. AB - The present study demonstrates that an operant conditioning paradigm-originally designed for assessing olfactory performance in an Old World primate, the pigtail macaque-can successfully be adapted for use with a New World primate, the spider monkey. Using a task based on a food-rewarded, two-choice discrimination of simultaneously presented odor stimuli, Ateles geoffroyi is capable of learning to discriminate between objects on the basis of odor cues. Moreover, animals could readily transfer to new S+ and S- stimuli, and could remember the significance of previously learned odor stimuli even after a 4-week break. We could also show that this method is suitable for obtaining reliable measures of olfactory sensitivity. Our results indicate that A. geoffroyi clearly outperforms Macaca nemestrina with regard to the speed of initial task acquisition and the ability to master transfer tasks, and shows a sensitivity to a food-related odorant which matches that of other primate species. PMID- 12576132 TI - Central insulin potentiates eating elicited by 2-deoxy-D-glucose. AB - 2-Deoxy-D-glucose (2DG) elicits glucoprivic food intake whether administered centrally or systemically. Insulin, on the other hand, elicits glucoprivic food intake when administered systemically but reduces food intake when administered centrally. The purpose of these experiments was to determine the interaction of centrally administered insulin with systemically administered 2DG on feeding. In the experimental condition, male Sprague-Dawley rats were administered 5 mU insulin into the third cerebral ventricle (i3vt) followed 2 h later by a subcutaneous injection of 250 mg/kg of 2DG. Contrary to expectations, third ventricular insulin significantly increased 2DG-induced hyperphagia. A replication using doses of insulin ranging from 1 to 10 mU revealed a dose dependent response. Whereas the lowest dose of insulin (1 mU) did not reliably change food intake, doses of 2.5, 5, and 10 mU significantly enhanced 2DG-induced feeding. Consistent with previous reports, centrally administered insulin, when given alone, caused a significant reduction of 24-h body weight and chow intake. To assess if the insulin-induced hyperphagia was a result of leakage from the ventricles, we peripherally administered 5 mU of insulin and observed, if anything, a slight decrease of food intake. These studies suggest that in the presence of central glucoprivation, a distinct anabolic action of centrally administered insulin overrides the normally observed catabolic response and increases the hyperphagic feeding response induced by 2DG. PMID- 12576133 TI - Temporal pattern of social aggregation in tadpoles and its influence on the measurement of lateralised response to social stimuli. AB - Tadpoles of several species have been proven to prefer using the left hemifield during fixation of their own mirror images. The lateral bias typically emerges some minutes after the placement of the animals in the test apparatus. Here we checked whether such a temporal pattern was associated with lateralisation per se, or rather reflected temporal variations in social aggregation. We tested the temporal changes in tadpoles' movements directed towards conspecifics and other parts of the environment. We found that the propensity to move to make social aggregation only appears after about 5 min following placement in a novel environment and this corresponded quite well with the appearance of lateralisation, when tadpoles showed a higher probability of approaching a conspecific appearing on their left hemifield rather than on their right hemifield. These findings confirm, using natural conspecifics, evidence that in tadpoles, the left hemifield is better at detecting and directing approach responses to social stimuli. PMID- 12576134 TI - Cardiovascular disorders associated with cocaine use: myths and truths. AB - Cocaine produces a pattern of cardiovascular responses that are associated with apparent myocardial ischemia, arrhythmias, and other life-threatening complications in some individuals. Despite recent efforts to better understand the causes of cocaine-induced cardiovascular dysfunction, there remain a number of unanswered questions regarding the specific mechanisms by which cocaine elicits hemodynamic responses. This review will describe the actions of cocaine on the cardiovascular system and the evidence for the mechanisms by which cocaine elicits hemodynamic and pathologic responses in humans and animals. The emphasis will be on experimental data that provide the basis for our understanding of the mechanisms of cardiovascular toxicity associated with cocaine. More importantly, this review will identify several controversies regarding the causes of cocaine induced cardiovascular toxicity that as yet are still debated. The evidence supporting these findings will be described. Finally, this review will outline the obvious deficits in our current concepts regarding the cardiovascular actions of cocaine in hope of encouraging additional studies on this grave problem in our society. PMID- 12576135 TI - Regulation of cardiac myocyte cell death. AB - Cardiac myocyte death, whether through necrotic or apoptotic mechanisms, is a contributing factor to many cardiac pathologies. Although necrosis and apoptosis are the widely accepted forms of cell death, they may utilize the same cell death machinery. The environment within the cell probably dictates the final outcome, producing a spectrum of response between the two extremes. This review examines the probable mechanisms involved in myocyte death. Caspases, the generally accepted executioners of apoptosis, are significant in executing cardiac myocyte death, but other proteases (e.g., calpains, cathepsins) also promote cell death, and these are discussed. The two principal cell death pathways (death receptor- and mitochondrial-mediated) are described in relation to the emerging structural information for the principal proteins, and they are discussed relative to current understanding of myocyte cell death mechanisms. Whereas the mitochondrial pathway is probably a significant factor in myocyte death in both acute and chronic phases of myocardial diseases, the death receptor pathway may prove significant in the longer term. The Bcl-2 family of proteins are key regulators of the mitochondrial death pathway. These proteins are described and their possible functions are discussed. The commitment to cell death is also influenced by protein kinase cascades that are activated in the cell. Whereas certain pathways are cytoprotective (e.g., phosphatidylinositol 3'-kinase), the roles of other kinases are less clear. Since myocyte death is implicated in a number of cardiac pathologies, attenuation of the death pathways may prove important in ameliorating such disease states, and possible therapeutic strategies are explored. PMID- 12576136 TI - Electrical stimulation of thalamic Nucleus Submedius inhibits responses of spinal dorsal horn neurons to colorectal distension in the rat. AB - In 78 halothane-anesthetized rats, we characterized the responses of single neurons in the dorsal horn of L(6)-S(1) spinal segments to a noxious visceral stimulus (colorectal balloon distension, CRD), and studied the effects of focal electrical stimulation of Nucleus Submedius (Sm) on these responses using standard extracellular microelectrode recording techniques. A total of 102 neurons were isolated on the basis of spontaneous activity. Eighty (78%) responded to CRD, of which 70% had excitatory and 30% had inhibitory responses. Neurons showed graded responses to graded CRD pressures (20-100 mmHg), with maximum excitation or inhibition occurring at 100 mmHg. Responses to noxious (pinch, heat) and innocuous (brush, tap) cutaneous stimuli were studied in 73 of the spinal dorsal horn neurons isolated. Fifty-seven (78%) of these neurons (46 CRD-responsive and 11 CRD-nonresponsive) had cutaneous receptive fields, of which 35 (61%) were small and ipsilateral, 14 (25%) were large and ipsilateral, 7 (12%) were large or small and bilateral, and 1 (2%) was small and contralateral. Sixty one percent of these neurons responded to both noxious and innocuous cutaneous stimulation, 35% responded only to noxious stimulation, and 4% responded only to innocuous stimulation. Electrical stimulation (50-300 microA) of the contralateral Sm produced intensity-dependent attenuation of the CRD-evoked activities of most neurons (18/28 of CRD-excited and 7/12 of CRD-inhibited) tested. Sm stimulation produced facilitation of CRD responses of only one neuron (CRD-inhibited). Sm stimulation had no effects on spontaneous activity. These data indicate that Sm may be involved in the descending inhibitory modulation of visceral nociception at the spinal level. PMID- 12576137 TI - Loss of dendritic connectivity in CA1, CA2, and CA3 neurons in hippocampus in rat under aluminum toxicity: antidotal effect of pyridoxine. AB - Aluminum chloride (AlCl(3); 4 mg/kg) was injected into the cerebrospinal fluid of adult rats as a one time dose. Rapid Golgi stained sections of hippocampus were examined for detailed histology of neurons in CA1, CA2, and CA3 areas. The axonal length and number of dendritic branches were seen reduced 30 days later in aluminum (Al)-injected group when compared to vehicle-injected controls. Of these perturbations, dendritic branches were seen reduced significantly. Al toxicity apparently affects neuronal connectivity in hippocampus. These perturbations are reversed by supplementing the feed with pyridoxine (8 mg/kg) for 30 days. As the loss of synaptic connectivity is a predominant feature of neurodegenerative disorders such as Alzheimer disease, this study may have implications in such disorders. Pyridoxine may be considered as a potent antidote to Al toxicity and neurodegenerative disorders such as Alzheimer disease. PMID- 12576138 TI - Masculine sexual activity affects slow wave sleep in Golden hamsters. AB - The sleep pattern is modified by events occurring during wakefulness. In rats, it has been shown that male sexual behavior has a direct influence on sleeping patterns, increasing slow wave sleep (SWS) duration. On the other hand, the sexual behavior pattern of the male Golden hamster differs from the copulatory pattern of male rats. Male hamsters copulate faster and they do not display the motor inhibition observed in rats after each ejaculation. Moreover, close to exhaustion, hamsters display a behavioral pattern known as Long Intromission, which has been linked to an sexual inhibitory process. The present study was performed to determine the effects of male sexual activity on the sleep pattern in hamsters. Subjects were allowed to copulate for 30 and 60 min. In addition, the effect of locomotor activity was also assessed. The results show that male sexual behavior induced a significant increase of SWS II, with a reduction of wakefulness. No effect was observed on REM sleep. Locomotor activity produced only a slight effect on sleep. The results are discussed in terms of the similarities between the effects observed after sexual behavior on sleep in rats and hamsters, despite the substantial differences in the behavioral pattern. PMID- 12576139 TI - Significance of endogenous opioids in the maintenance of cerebral and spinal vascular CO2-sensitivity in deep hemorrhagic hypotension. AB - High CO(2)-sensitivity, one of the major characteristics of the cerebrovascular bed, has been shown to be influenced by a variety of factors. There are no reports, however, on the involvement of the endogenous opioid peptides in the modulation of the CO(2)-sensitivity of the cerebral and spinal cord vessels, either in normotensive or, in hypotensive conditions. The effect of general opiate receptor blockade (1.0mg/kg naloxone, i.v.) on regional cerebrovascular CO(2)-sensitivity was studied with radiolabeled microspheres in 10 distinct brain and spinal cord regions of the anesthetized cat. The CO(2)-induced flow changes were investigated in normotensive, in moderately hypotensive (MAP=80 mmHg) and in deep hypotensive cats (MAP=40 mmHg). The systemic arterial pressure was lowered by hemorrhage. In the normotensive cats, opiate receptor blockade caused no changes in the vascular CO(2)-sensitivity in the investigated cerebral and spinal cord regions. In moderate hypotension, cerebral and spinal CO(2)-sensitivity was significantly reduced by the hemorrhage itself, but remained unaffected by the naloxone administration. In deep hemorrhagic hypotension, however, general opiate receptor blockade resulted not only in a further reduction of the already impaired CO(2)-sensitivity, but even in a reversal of the effect of CO(2) from flow increase to flow decrease. These results indicate that endogenous opioid peptides, which do not seem to influence cerebrovascular reactions in steady state, normotensive conditions, may contribute significantly to the maintenance of the normal vasodilatory response of the cerebral and spinal cord vessels to CO(2) during hemorrhage-induced deep arterial hypotension. PMID- 12576140 TI - Coactivation in arm and shoulder muscles during voluntary fixation of a single joint. AB - The objective of this study was to determine the organization of coactivation in the arm and shoulder muscles. Normal human subjects made alternate movements of a joint in the horizontal plane, either adduction-abduction of the second finger and shoulder, ulnar-radial deviation of the wrist, or extension-flexion of the elbow, during which they fixed a focal joint while decreasing the movement amplitude and increasing the fixation strength. They varied the fixation strength at four different levels up to the maximum. The focal-joint angle, and surface electromyograms (EMGs) from the intrinsic hand, antebrachial, upper-arm, and shoulder muscles were recorded. EMGs in the phase of fixation were quantified by integration after rectification. The degree of coactivation among the muscles was evaluated by calculating correlation coefficients across the integrated EMGs. There were correlations in the integrated EMGs among focal-joint muscles (FJMs), and also between one of the FJMs and the muscles distal and/or proximal to the FJMs: in the finger fixation between the hand and antebrachial muscles, in the wrist fixation between the antebrachial and hand/upper-arm muscles, in the elbow fixation between the upper-arm and antebrachial/shoulder muscles, and in the shoulder fixation between the shoulder and upper-arm muscles. Moderate or slight correlations were seen in muscles more distant from FJMs. Our results indicate that the longitudinal distance from FJMs in the shoulder and arm muscles is an important factor in determining levels of coactivation. This is discussed in relation to the fact that neighboring muscles share joints with FJMs. PMID- 12576141 TI - Endothelial activation is an intermediate step for peripheral lipopolysaccharide induced activation of paraventricular nucleus. AB - Peripheral injection of bacterial endotoxin lipopolysaccharide (LPS) activates the paraventricular nuclei of the hypothalamus (PVN), and consequently the hypothalamus-pituitary adrenal axis. Inflammatory cytokine interleukin-1 (IL-1) has been considered as a key mediator that translates the peripheral LPS stimulation into neuronal activation in the PVN. Several studies attempting to localize the expression of receptors for IL-1 (IL-1R), however, have failed to detect IL-1R on PVN neurons. It remains unclear, therefore, how IL-1 might stimulate the neurons of the PVN. In this study, we traced the cellular responsiveness to IL-1 by measuring the mRNA production of the cytokine responsive gene IkappaBalpha in the PVN. After either peripheral injection LPS or intracerebroventricular (i.c.v.) injection of IL-1beta, IkappaBalpha mRNA was found mostly in endothelial cells of the brain with the highest level of expression in PVN blood vessels. In addition, both injections also induced the expression of cyclooxygenase-2 in brain endothelial cells. Pretreatment with indomethacin, a cyclooxygenase inhibitor, blocked LPS and IL-1 induced neuronal activation in the PVN, but did not reduce the induction of IkappaBalpha in PVN endothelium. These results show that IL-1 acting on the endothelial cells of the brain, particularly in the PVN, may be an intermediate step relating peripheral immune signals to the brain. PMID- 12576142 TI - Oxypurinol administration fails to prevent hypoxic-ischemic brain injury in neonatal rats. AB - The purpose of the present study was to determine whether oxypurinol, a xanthine oxidase inhibitor, reduces free radicals and brain injury in the rat pup hypoxic ischemia (HI) model. Seven-day-old rat pups had right carotid arteries ligated followed by 2.5h of hypoxia (8% oxygen). Oxypurinol or vehicle was administered by i.p. injection at 5 min after reoxygenation and once daily for 3 days. Brain damage was evaluated by weight deficit of the right hemisphere at 22 days following hypoxia. Oxypurinol treatments did not reduce weight loss in the right hemisphere. Brain weight loss in the right hemisphere were -26.2+/-3.6, -15.2+/ 6.9, -21.7+/-4.4, -15.8+/-5.1, and -16.7+/-3.4% in vehicle (n=33), 10 (n=17), 20 (n=16), 40 (n=15), and 135 mg/kg (n=13) oxypurinol-treated groups (p>0.05), respectively. Brain thiobarbituric acid-reacting substances (TBARS) were assessed 3 and 6h after reoxygenation. Concentrations of TBARS rose 1.5-fold due to HI. Oxypurinol did not significantly reduce an HI-induced increase in brain TBARS. Thus, xanthine oxidase may not be the primary source of oxy-radicals in pup brain and as such oxypurinol does not prevent free radical-mediated lipid peroxidation or protect against brain injury in the neonatal rat HI model. PMID- 12576143 TI - Examination of sensorimotor performance following middle cerebral artery occlusion in rats. AB - Middle cerebral artery occlusion (MCAO) in rats is the most commonly used stroke model. Besides the infarct size, assessment of sensorimotor performance has become increasingly important in neuroprotective drug research. However, contradictions exist about procedures for testing functional outcome following MCAO. The aim of the present study was to evaluate a relatively simple set of neurological tests based on the most commonly used scoring systems, and to describe the functional recovery and correlation with the infarct size in rats sacrificed 2 or 14 days after permanent or transient MCAO. The smaller infarct size of rats with transient occlusion was reflected in the neurological scores only during the first 6h. By day 14, no recovery occurred in postural signs, lateral resistance and spontaneous activity, other signs showed different degrees of recovery. Correlation with the infarct size was found only on certain days in gait disturbance, placing reactions, daily body weight and spontaneous activity. According to our observations, the most commonly used sensorimotor tests provide a useful initial screening of functional deficits, but these tests most probably measure deficits caused by infarction of the core area. It is suggested that these tests should be completed by more refined tests when testing a neuroprotective drug which reduces the infarct size in penumbral areas. PMID- 12576144 TI - Effects of arginine-vasopressin (AVP) on long-term potentiation in intact anesthetized rats. AB - We studied the effects of the neuropeptide arginine-vasopressin (AVP) on the long term potentiation (LTP) paradigm in the dentate gyrus (DG) of urethane intact anesthetized rats. Intracerebroventricular injection of 1 microg of the hormone in 1 microl of physiological solution 3 min before tetanization, produced a significant increase in both components of the perforant path-evoked potentials (EP) in the DG. The effects were already evident 1 min after tetanization. Amplitude of the EPs increased continuously for the 2h of recording time, reaching values 100% above baseline, reference levels. In contrast, in previous in vitro studies, enhancement of LTP with AVP appeared only after 15 min of exposure of the hippocampal slice to the hormone, increased EPSPs were no higher than 50% from baseline, reached a plateau at 40 min decreasing slowly thereafter. Not only quantitative but also qualitative differences can be observed between in vitro and in vivo intact preparations in response to identical hormones. This study emphasizes the importance of hormone neurotransmitter interactions in determining electrophysiological characteristics of response to AVP. PMID- 12576145 TI - Ultrastructure of ascending cholinergic terminals in the anteroventral thalamic nucleus of the rat: a comparison with the mammillothalamic terminals. AB - In this study, to identify the ultrastructure and distribution of ascending cholinergic afferent terminals in the anteroventral thalamic nucleus, we used an anti-vesicular acetylcholine transporter antibody as marker of cholinergic afferents, and characterized the immunoreactive terminals at the ultrastructural level. We then compared the distribution pattern of the cholinergic terminals and that of the mammillothalamic terminals identified by anterograde transport of a tracer injected into the mammillary body. The cholinergic terminals were small, and formed both symmetrical and asymmetrical synaptic contacts throughout the dendritic arborizations, particularly in the distal region. This distribution pattern differed from that of mammillothalamic terminals, that were of LR (large terminal containing round synaptic vesicles) type and were preferentially distributed in the proximal region of dendrites. We also found relatively numerous cholinergic terminals making contact directly with immunonegative excitatory terminals, both LR and SR (small terminal containing round vesicles) terminals, without clear postsynaptic specialization. A few cholinergic terminals even seemed to form a synaptic complex with the LR or SR terminals. These findings suggest that the ascending cholinergic afferents in the anteroventral thalamic nucleus can effectively modulate excitatory inputs from both the mammillothalamic and corticothalamic terminals, in close vicinity to a synaptic site. PMID- 12576147 TI - Effects of pedunculopontine tegmental nucleus lesions on emotional reactivity and locomotion in rats. AB - Bilateral damage to the pedunculopontine tegmental nucleus (PPTg) has been found to impair several learning tasks; however, it is not clear whether this effect could be at least partially attributable to changes in the rat emotional reactivity and/or spontaneous locomotion. Therefore, the present work has tested the effects of bilateral electrolytic lesions of the PPTg on the behaviour of rats in the elevated plus-maze and the open field test. Because the behaviour of rats in learning and emotional tasks can be sensitive to routine experimental manipulations, we also have tested the effects of brief pre-surgical handling procedures on anxiety-like behaviours and locomotion in both lesioned and control rats. Lesions of the pedunculopontine tegmental nucleus (1). did not have any effects on spontaneous locomotor activity and (2). did not increase emotional reactivity. In fact, there was a slight bias towards a reduction in anxiety-like behaviours in lesioned rats, as evidenced by a significant increase in the number of open arm entries. Pre-surgical handling induced a slight decrease of emotional reactivity and a slight increase of exploratory activity. We conclude that damage to the pedunculopontine tegmental nucleus is not accompanied by either an enhancement of emotional reactivity or by an altered spontaneous locomotion. PMID- 12576146 TI - Non-competitive NMDA receptor antagonists moderate seizure-induced c-fos expression in the rat cerebral cortex. AB - We examined the effects of non-competitive NMDA glutamate receptor antagonists on seizures elicited by 4-aminopyridine (4-AP), and in particular, on the expression of the transcription factor c-fos induced by these seizures. Induction of c-fos mRNA due to 4-AP-elicited seizures was ascertained by reverse transcription polymerase chain reaction in samples of the neocortex. Adult rats were pretreated with the NMDA receptor antagonists amantadine (40 mg/kg), ketamine (3mg/kg), dizocilpine (MK-801; 1mg/kg) or dextrometorphan (40 mg/kg); 4-AP (5mg/kg) was then injected i.p. Controls were treated with either antagonist only or with 4-AP only. Pretreatment with the antagonists (with the exception of amantadine) increased the latency of behavioural seizures, but not all of the antagonists caused symptomatic seizure protection. In the brains which were processed for Fos immunohistochemistry, quantitative evaluation of immunostained cells was performed in the neocortex and hippocampus. Treatment with either antagonist did not induce by itself c-fos expression, with the exception of amantadine, which caused slight Fos induction in the neocortex. Pretreatment with all the antagonists resulted in decrease of seizure-induced Fos immunoreactivity with respect to non-pretreated animals. Decrease of immunostained cells was significant in the neocortex, in the granule cell layer and hilus of the dentate gyrus, in hippocampal areas CA1 and CA2. MK-801, ketamine and dextrometorphan decreased significantly Fos immunoreactivity also in area CA3. The decrease of Fos immunostaining was not directly correlated with a suppression of behavioural seizures. The results support an important role of NMDA receptors in c-fos gene induction in acute 4-AP seizures. PMID- 12576148 TI - Atrial natriuretic peptide mediates oxytocin secretion induced by osmotic stimulus. AB - Atrial natriuretic peptide (ANP), first discovered in the heart, has been also detected in various brain regions involved in the control of cardiovascular function and water and sodium balance. The anteroventral region of the third ventricle (AV3V) and the subfornical organ (SFO) have ANP-immunoreactive projections towards the paraventricular (PVN) and supraoptic (SON) nuclei of the hypothalamus. Extracellular fluid (ECF) hyperosmolality stimulates the secretion of oxytocin (OT) which induces ANP release by the atrium. On the other hand, passive immunoneutralization of ANP reduces OT secretion in response to ECF hypertonicity. Previous studies have shown the co-localization of ANP and OT in PVN and SON neurons and in the periventricular region, as well as the presence of ANPergic and oxytocinergic neurons in the median eminence. The aim of the present study was to investigate the OT and ANP content in the SON and PVN of the hypothalamus and in the posterior pituitary (PP) after an osmotic stimulus that induces OT secretion. The results showed that intracerebroventricular microinjection of normal rabbit serum (NRS) or of ANP antiserum followed or not by an intraperitoneal injection of isotonic saline did not alter OT secretion or OT content in the PVN, SON, and PP; passive ANP immunoneutralization reduced the basal content of ANP in the PVN, SON, and PP of animals in a situation of isotonicity; the ANP antiserum inhibited the increase of OT secretion and content of OT and ANP in the PVN, SON and PP induced by the osmotic stimulus. Thus, the increase in plasma OT and oxytocinergic neurons of the hypothalamus-posterior pituitary system in response to hypertonicity depends on the action of endogenous ANP, i.e., ECF hypertonicity must activate ANPergic neurons which directly or indirectly stimulate OT release. PMID- 12576149 TI - Potentiation of amphetamine-induced changes in dopamine and 5-HT by a 5-HT(6) receptor antagonist. AB - Although recent data has shown that 5-HT(6) receptor antagonists' can enhance basal cholinergic and glutamatergic neurotransmission in the cortex and hippocampus, the distribution of this receptor within terminal regions of the dopaminergic system suggests a possible role for this receptor in the modulation of dopamine (DA). Therefore, the role of the 5-HT(6) receptor was examined in the rat striatum in the presence and absence of the DA transport inhibitor/releaser, amphetamine. Amphetamine (0.3mg/kg s.c.) induced a selective increase in extracellular DA reaching a maximum of 311.3+/-73.5% of preinjection levels. Administration of SB-271046 (1 and 10mg/kg s.c.) followed by amphetamine produced an augmentation of amphetamine-induced changes in both DA and 5-hydroxytryptamine (5-HT), reaching maximum levels of 510.1+/-110.5% and 271+/-93.4% of preinjection values, respectively. Similarly, local infusion of amphetamine (100 nM) resulted in an increase in striatal DA levels reaching a maximum of 365.7+/-73.3% of preinfusion values. However, combination treatment with SB-271046 (1mg/kg s.c.) and amphetamine produced no augmentation of amphetamine-induced increases in extracellular levels of DA or in any other neurotransmitter measured. Taken together these data indicate that the 5-HT(6) receptor is not playing a role in the tonic modulation of NA, DA, 5-HT or glutamate neurotransmission in the striatum. However, when dopaminergic neurotransmission is enhanced the 5-HT(6) receptor appears to have a modulatory influence on not only DA but also 5-HT systems. This is the first direct neurochemical evidence that the 5-HT(6) receptor may have modulatory influences on both DA and 5-HT systems in the rat striatum. PMID- 12576150 TI - Neuronal endosomal/lysosomal membrane destabilization activates caspases and induces abnormal accumulation of the lipid secondary messenger ceramide. AB - Impairment of endosomal/lysosomal functions are reported as some of the earliest changes in several age-related neurological disorders such as Alzheimer's disease. Dysregulation of the lysosomal system is also accompanied by the accumulation of age-associated pigments and several recent reports have indicated that this age-related lipofuscin accumulation can sensitize cells to oxidative stress and apoptotic cell death. In this study, we have established and evaluated an in vitro age-related pathology paradigm that models lipofuscin accumulation. Our model consists of the treatment of cultured primary mouse neurons with lysosomotropic detergents. We have observed that one of the earliest biochemical changes associated with lysosomotropic detergent-induced membrane instability is a loss of the endosomal/lysosomal proton gradient integrity, followed by an activation of sphingomyelin hydrolysis and ceramide accumulation within enlarged endosomal/lysosomal vesicles. In addition, we demonstrate that ceramide accumulation correlates with the activation of proximal procaspases-8 and -9 as well as distal caspase-3, prior to the appearance of cell death. Taken together, we propose that disturbances of the endosomal/lysosomal system, in addition to the activation of the sphingomyelinase hydrolysis cycle, play essential roles in the course of post-mitotic neuronal aging. The abnormal accumulation of undigested lipids and proteins within dysfunctional endosomal/lysosomal vesicle populations during the process of pathological aging may serve as triggers of the cell death programs that are associated with downstream neurodegeneration. PMID- 12576151 TI - Psychophysical and EEG responses to repeated experimental muscle pain in humans: pain intensity encodes EEG activity. AB - Clinical pain is often characterized by repetitive and persistent occurrence in deep structures, but few studies investigated repetitive tonic pain in humans. To determine cerebral responses to repetitive tonic pain, psychophysical responses, and electroencephalographic (EEG) activation to five trials of repeated tonic muscle pain induced by hypertonic saline were examined and analyzed in 13 male subjects. The study was composed of two experimental sessions performed in separate days. Five sequential injections of hypertonic saline (5.8%) were used to induce repeated muscle pain in the left forearm, and five sequential injections of isotonic saline (0.9%) acted as control. Visual analogue scales (VAS) for pain intensity and 32-channels EEG activities were recorded simultaneously. Five trials of relatively stable muscle pain were induced by intramuscular injections of hypertonic saline, but no evident pain was induced by the injections of isotonic saline. Significant decreases in alpha-1 and -2 activities in posterior part of the head were found during repeated muscle pain in comparison with non-pain. In comparison with baseline, alpha-1 and -2 activities reduced significantly during the first two trials, and gradually resumed in the following three trials of muscle pain. However, beta-2 activity increased consistently throughout the five trials of muscle pain compared to baseline. Alpha-1 activity was negatively, but beta-2 activity was positively correlated to the pain intensity and pain area on the skin. Throughout five injections, the reduction of alpha-1 activity was contrary to the changes of pain intensity. These results indicates that pain-related EEG activities were encoded by the pain intensity. The thalamo-cortical system and descending inhibitory neuronal networks may be involved in the regulation of pain intensity. PMID- 12576152 TI - Are there biofilm-specific physiological pathways beyond a reasonable doubt? AB - The development of surface-attached communities called biofilms is often thought to be a regulated developmental process involving profound biofilm-specific modifications. Despite intense scrutiny and a growing body of evidence correlating changes in gene expression with the switch from planktonic-to-biofilm lifestyle, an integrated view of biofilm formation is still missing. PMID- 12576153 TI - Klebsiella pneumoniae type 3 pili facilitate adherence and biofilm formation on abiotic surfaces. AB - The role of type 1 and type 3 pili in mediating Klebsiella pneumoniae colonization of inert surfaces was assessed by analysis of 26 clinical, 7 water distribution system and four reference strains. Amongst the 33 wild type strains tested, 57.6% expressed type 1 pili, 54.5% expressed type 3 pili, 42.4% were highly hydrophobic, 69.7% were highly adherent to both glass and polypropylene surfaces, and 48.5% were efficient biofilm-forming strains on polystyrene dishes. High adherence and biofilm formation were positively correlated with bacterial surface hydrophobicity, type 3 fimbriae expression but not with type 1 pili expression and were not dependent upon the strain's origin. The K. pneumoniae CF3051 reference strain expressing only type 1 fimbriae adhered slightly to glass and polypropylene and did not form biofilm on polystyrene. K. pneumoniae IA565 and CF3097 reference strains producing type 1 and type 3 fimbriae showed efficient adherence to both glass and polypropylene and biofilm formation on polystyrene. Moreover, transformation of the afimbriated, weakly-adherent CF3172 strain with the recombinant pFK10 plasmid carrying the mrk gene cluster resulted in type 3 fimbriae expression, increased surface hydrophobicity, increased adherence to abiotic surfaces and biofilm formation. Thus, type 3 pili constitute the main K. pneumoniae adhesive factor, facilitating adherence and biofilm formation on abiotic surfaces of strains of different origins. PMID- 12576154 TI - Degradation of 4-amylphenol and 4-hexylphenol by a new activated sludge isolate of Pseudomonas veronii and proposal for a new subspecies status. AB - Novel Pseudomonas strains INA04, INA05, and INA06, were isolated from activated sludge. Strain INA06 was found to degrade long chain alkylphenols such as 4-n amylphenol and 4-n-hexylphenol as the sole source of carbon, apart from co metabolic degradation of 4-n-nonylphenol in the presence of phenol, while INA04 and INA05 could grow on phenol, but could not grow well on alkylphenols. Induction studies on strain INA06 revealed a broad substrate-specific phenol hydroxylase, for the metabolism of phenol and alkylphenols, inducible with phenol or para-substituted alkylphenol. They bore close resemblance to members of Pseudomonas sensu stricto. 16S rDNA sequence homology of INA06 was closest to P. veronii (99.7%). DNA-DNA hybridization pointed out higher linkage (64% similarity) to the type strain of P. veronii than to other species of Pseudomonas sensu stricto (>60%). The BOX-PCR profile of all INA strains was similar, but different from that of P. veronii. Since biochemical characteristics were similar to those of P. veronii, and genetic relatedness was at the margin of species differentiation level (70%), we propose these strains to be treated as a new subspecies of P. veronii. The type strain of this new subspecies, named P. veronii subsp. inensis subsp. nov., is strain INA06. The accession number of strain INA05 is CIP 107595=JCM11829, and that of INA06 is CIP107594(T)=JCM11828(T). The 16S rDNA sequence accession number (DDBJ/EMBL/GenBank) of strain INA06 is AB056120. PMID- 12576155 TI - Ribosomal DNA sequence analysis shows that the basidiomycete C30 belongs to the genus Trametes. AB - The basidiomycete C30 was considered as an isolate of a population of Marasmius quercophilus collected on evergreen oak litter from the Mediterranean forest. Recent phenotypic studies have clearly shown that it differs from newly characterized M. quercophilus isolates. Subsequent analysis of laccase genes revealed that C30 sequences are similar to laccase encoding sequences from organisms belonging to the polyporoid clade. Comparison of sequences of the C30 ITS regions, including 5.8S rDNA, with those found in databanks confirmed that C30 is not a Marasmius. Finally, 25S rDNA analysis revealed that C30 is closely related to the Coriolaceae and, in particular, to Trametes trogii. PMID- 12576156 TI - Hemolytic activity of Prevotella intermedia and Prevotella nigrescens strains: influence of abiotic factors in solid and liquid assays. AB - The influence of growth medium, hemin and menadione, blood source and atmosphere of incubation on the expression of hemolytic activity of 25 strains of Prevotella intermedia and Prevotella nigrescens was evaluated. The best hemolytic activity was observed for samples of both species growing in brain heart infusion agar and incubated in Brewer-like anaerobic jars for 48 h. Hemolysis was less intense and occurred later in the presence of hemin and menadione in solid media. beta Hemolysis was detected for medium supplemented with horse or human blood and alpha-hemolysis was observed when sheep blood was used. These results suggesting some specificity for the hemolytic activity were also observed in liquid assays in which sheep erythrocytes were found to be resistant to hemolysis while horse and human cells where lysed. In liquid assays, the hemolytic activity of all studied strains remained stable in the pH range of 6.0 to 8.5 and was not altered by iron-scavenging compounds or atmosphere of incubation. The phenomenon of hot/cold hemolysis was ruled out as the mechanism of action of P. intermedia and P. nigrescens hemolysin. PMID- 12576157 TI - Positive effects of growth at suboptimal temperature and high salt concentration on long-term survival of Lactobacillus sakei. AB - Lactobacillus sakei is a lactic acid bacterium commonly found on fresh meat and represents the predominant flora of vacuum-packed meat. In the present article, we studied the behavior of L. sakei in a chemically defined medium under various growth conditions relative to temperature or NaCl concentration. Growth occurred at each temperature, but growth rate and final cell density decreased at low temperature and survival was enhanced. In the presence of NaCl, we also observed a longer doubling time and a lower final cell density together with an enhanced long-term survival. When both conditions were combined, the long-term survival was greatly increased to about 28 weeks. Differences in cell morphology were observed under electron microscopy when cells were grown at low temperature in the presence of salt. We speculate that these are due to modifications in membrane structure. These results show that L. sakei is able to adapt to these environmental conditions and that slow growth is clearly associated with enhanced long-term survival. PMID- 12576158 TI - Stress response in Actinobacillus actinomycetemcomitans: induction of general and specific stress proteins. AB - Stress proteins are highly conserved proteins that are essential for cell survival. In this study, the induction of general and specific stress proteins in Actinobacillus actinomycetemcomitans cells subjected to different stress conditions was evaluated by two-dimensional SDS-PAGE analysis. Twenty-eight (up- or down)regulated proteins, including DnaK and GroEL proteins, were identified as general stress proteins. In addition, eighteen regulated proteins were classified as pH stress-specific proteins, ten as acid stress-specific proteins, five as alkaline stress-specific proteins, three as heat stress-specific proteins, and ten as acid/heat stress-specific proteins. Further proteomic studies are required to determine the exact nature of the proteins regulated during the stress response of A. actinomycetemcomitans. PMID- 12576159 TI - Nutritional and cultural parameters influencing antidipteran delta-endotoxin production. AB - In this study, various nutritional and cultural parameters influencing diptera specific delta-endotoxin synthesis by Bacillus thuringiensis subsp. israelensis HD500 were investigated. Of various inorganic nitrogen sources, the highest yields of Cry11Aa and Cry4Ba proteins were obtained on (NH(4))(2)HPO(4). Among carbon sources, inulin, dextrin, maltose, lactose, sucrose, whey and glycerol were all stimulatory, while glucose, starch and molasses were suppressive. High concentrations of inorganic phosphate (50 to 100 mM K(2)HPO(4)) were required for an effective synthesis of Cry4Ba. Mn was the most critical element for the biosynthesis of both toxins at 10(-6) M concentration. Mg and Ca favored production when provided at 8 x 10(-3) M and 5.5 x 10(-4) M concentrations, respectively, while Fe, Zn and Cu negatively influenced biosynthesis. Cry4-toxin synthesis was best at neutral pH and also when the organism was grown at 25 degrees C. Throughout the study, the extent of growth and sporulation of the producer organism was also monitored. PMID- 12576160 TI - Plasmodium immobilization of Physarella oblonga (Berk. & Curt.) Morgan (Myxomycetes) using kaolinite as a matrix of entrapment. AB - The possibility of using kaolinite-immobilized plasmodium fragments of Physarella oblonga (Berk. & Curt.) Morgan to maintain their metabolic activity was examined. The immobilization process was carried out with 1 mg of plasmodium of P. oblonga entrapped in 10 g of kaolinite. Sodium acetate (1 mM) was used as a metabolic precursor. The collection of fractions was carried out during a one month period, and extracted with ether/ethyl acetate and chloroform/acetonitrile. The extractions from plasmodium in natura were accomplished with the same solvents. The extracts obtained were analyzed in a spectrophotometer at 266 nm and 310 nm, and by thin layer chromatography to assess the productivity of the immobilized plasmodium. The absorbances of the extracts in both wavelengths and the chromatographic tests showed the synthesis of compounds by the immobilized material. Three chromatographic spots were observed in the extracts obtained from the immobilized plasmodium. Two spots coincided with the R(f) values and coloration of the spots observed for the material in natura used as a reference. The kaolinite-immobilized plasmodium of P. oblonga can remain metabolically active for at least one month at room temperature and ambient light conditions. PMID- 12576161 TI - Development of strain-specific PCR reactions for the detection of the probiotic Escherichia coli strain Nissle 1917 in fecal samples. AB - PCR was used to establish a specific detection system for the non-pathogenic Escherichia coli strain Nissle 1917 (DSM6601), which is used as a probiotic drug against intestinal disorders and diseases. Five PCR assays have been developed which are based on the chromosomally encoded major fimbrial subunit genes fimA (type 1 fimbriae) and focA (F1C fimbriae), and the two small cryptic plasmids pMUT1 and pMUT2. The assays were validated by testing a collection of 354 different pathogenic and non-pathogenic E. coli strains from various origins, including E. coli K-12, fecal and environmental as well as pathogenic extraintestinal and intestinal E. coli strains. The most specific results were obtained with primers based on DNA sequences from plasmid pMUT2. The plasmid based PCR assays described can be used to detect E. coli strain Nissle 1917 in feces from patients without prior cultivation. PMID- 12576163 TI - Current limitations of antiepileptic drug therapy: a conference review. AB - The current limitations of antiepileptic drug (AED) therapy were the topic of a discussion group meeting at the 5th European Congress on Epileptology, Madrid, 6 10 October 2002. This review contains four short papers covering the topics discussed by the speakers at this meeting and an account of the ensuing discussion with all participants. The meeting focused on four issues. (i) Are mechanisms of action of AEDs useful to predict treatment outcome? The short answer to this question was no, for several reasons. These include the fact that clinically relevant mechanisms in individual patients remain unclear, the treatment of epilepsy targets the symptoms rather than the cause of the disease, and that current seizure classification defines heterogeneous patient populations. (ii) The benefits of the often recommended titration of the dose to the maximum tolerated level when seizures persist at average AED doses. A re evaluation of this practice showed that dose escalation achieves seizure freedom in only 1 of 4 patients with newly diagnosed epilepsy and only 1 of 10 patients with refractory epilepsy are likely to experience a greater than 50% reduction in seizure frequency. Being aware of the limited utility of maximum dose titration and subsequent dose reduction if no significant individual benefit is achieved avoids medical over-treatment with a worsening risk-benefit balance. (iii) When single drug therapy is not sufficiently effective, adding a second drug or alternative monotherapy are common options. Based on published data, there is no conclusive evidence in favour of either alternative monotherapy or second-line polytherapy. A pragmatic choice may be to evaluate the combination and then attempt to withdraw the first drug in the case of success. This may prevent the substitution of a partially efficacious drug by a non-efficacious drug. The choice of the second drug should, in theory, be based on which first drug has failed but again compelling evidence to support specific recommendations is lacking. (iv) Unexpected worsening of seizures may occur in many circumstances and has many causes, including tolerance and adverse pharmacodynamic effects of individual AEDs on seizure generating mechanisms. Patients are usually aware of aggravation and may express a "dislike" for a particular AED as a warning sign for physicians to modify the medication. The availability of numerous AEDs, particularly with single mechanisms of action, has increased the risk of paradoxical effects that may go undetected in clinical trials and only surface during astute clinical observations. PMID- 12576164 TI - Valproate unbound fraction and distribution volume following rapid infusions in patients with epilepsy. AB - The availability of an intravenous formulation now makes possible rapid administration of valproate (VPA) loading doses, but estimates of key VPA pharmacokinetic parameters in patients have limited the use of this approach. VPA disposition was characterized in 112 epilepsy patients, with or without enzyme inducing comedications, randomized to either 3.0 or 1.5mg/kg/min infusions of valproate sodium injection. Maximum dose was 14. During the first week of life, KCNQ-channels performed as a predominant inhibitory system, whereas after this period GABAergic-transmission switched from excitatory to inhibitory function. Contrary, glutamatergic-transmission has acquired as excitatory function from P0. These findings suggest that the pathogenic mechanisms of age dependent development and spontaneous remission of BFNC are, at least partially, associated with the interaction between age-dependent reduction of inhibitory KCNQ-channel activity and age-dependent functional switching of the GABAergic system from excitatory to inhibitory action in neonatal CNS. PMID- 12576171 TI - Effects of lamotrigine and levetiracetam on seizure development in a rat amygdala kindling model. AB - In kindling models of epilepsy, the period during which repeated stimulation evokes intensifying seizures is attributed to an underlying epileptogenic process, and the point at which class 5 kindled seizures occur is considered the established epileptic state. Previous studies have indicated that a separation can occur between drug effects on these two components. For example, carbamazepine and phenytoin inhibit kindled seizures but have no effect on seizure development, whereas levetiracetam inhibits both components. We have investigated the profile of lamotrigine in the amygdala kindling model, including levetiracetam for comparison. As expected, both treatments dose-dependently inhibited class 5 kindled seizures. In a separate study, daily administration of either lamotrigine (20mgkg(-1) i.p.) or levetiracetam (50mgkg(-1) i.p.) demonstrated antiepileptogenic-like effects by blocking seizure development during the treatment period. Following cessation of drug treatment, further daily stimulation resulted in kindled seizure development, though there was a significant increase with both treatment groups, relative to the control group, in the total number of stimulations required to produce classes 3 and 5 seizures. In addition, prior levetiracetam treatment appeared to delay or prevent the expected increase in after-discharge duration (ADD). These results suggest that lamotrigine, like levetiracetam, possesses the ability to counteract kindling acquisition, which differentiates it from other drugs with sodium channel blocking activity. PMID- 12576172 TI - Functional characterization of the D188V mutation in neuronal voltage-gated sodium channel causing generalized epilepsy with febrile seizures plus (GEFS). AB - Mutations in the alpha 1 subunit of the voltage-gated sodium channel (SCN1A) have been increasingly recognized as an important cause of familial epilepsy in humans. However, the functional consequences of these mutations remain largely unknown. We identified a mutation (D188V) in SCN1A segregating with generalized epilepsy with febrile seizures (GEFS) in a large kindred. Compared to wild-type sodium channels, in vitro expression of channels harboring the D188V mutation were found to be more resistant to the decline in amplitude that is normally observed over the course of high frequency pulse trains. This small change on a single aspect of channel function is compatible with an increase in membrane excitability, such as during sustained and uncontrolled neuronal discharges. These data suggest that this specific effect on sodium channel function could be a general mechanism in the pathophysiology of epilepsies caused by mutations in sodium channels in humans. PMID- 12576173 TI - The protective effect of a ketogenic diet on kainic acid-induced hippocampal cell death in the male ICR mice. AB - This study was designed to evaluate the antiapoptotic effects of a ketogenic diet (KD) through histological (cresyl violet staining, TUNEL staining and immunohistochemistry) and behavioral studies using kainic acid (KA, 25mg/kg i.p.) induced seizures in male ICR mice. KA-induced seizure in rodents is widely used as an experimental model for human temporal lobe epilepsy because of their behavioral and pathological similarities. A KA-induced seizure causes neuronal damage in hippocampal pyramidal neurons and involves a caspase-3-mediated apoptotic pathway. In this study, the seizure onset time of the KD-fed group was delayed compared to that of the group fed a normal diet (ND) after a systemic KA injection. Histological studies revealed that KA caused pyknosis in most of the hippocampal areas in the ND-fed group, however, well-preserved pyramidal neurons were detected in the hippocampus of mice that had been on KD for 1 month, which began on postnatal day 21. The number of TUNEL-positive cells and caspase-3 positive cells in the hippocampus of the KD-fed group was lower than that of the ND-fed group. These findings indicate that KD has an antiepileptic effect via a neuroprotective action that involves the inhibition of caspase-3-mediated apoptosis of hippocampal neurons. PMID- 12576174 TI - NMDA-induced seizures in developing rats cause long-term learning impairment and increased seizure susceptibility. AB - N-methyl-D-aspartate (NMDA) receptors play a prominent role in the pathogenesis of epilepsy, yet few studies have used NMDA as a convulsant in whole animals. In developing rats, systemic NMDA induces seizures with a unique seizure phenotype ("emprosthotonic" or hyperflexion seizures) and electrographic pattern (electrodecrement). These features are not seen in kainic acid-induced seizures, suggesting that seizures activated by NMDA might cause different long-term consequences. Therefore, we investigated the effects of NMDA seizures during development on cognitive function and susceptibility to seizures in adulthood. Rat pups (P12-20) were injected with saline (n=36) or NMDA (n=64) at convulsant doses (15-30mg/kg, i.p.). After NMDA injection, a characteristic sequence of seizure activity was seen: initial behavioral arrest, followed by hyperactivity, agitation, and then emprosthotonus and generalized tonic-clonic seizures. Seizures were terminated 30min later by ketamine (50mg/kg, i.p.). On P85, rats underwent behavioral testing in the water maze. Rats that had experienced NMDA seizures as pups took significantly longer to learn the platform location over 5 days of testing, compared to controls. On P90, rats were injected with pentylenetetrazol (PTZ, 50mg/kg, i.p.) to assess their susceptibility to generalized seizures. NMDA-treated rats had decreased latency and increased duration of class V PTZ seizures. Cresyl violet-stained sections of cortex and hippocampus had no obvious cell loss or gliosis. In summary, NMDA causes a unique seizure phenotype in the developing brain, with subsequent deficits in spatial learning and an increased susceptibility to PTZ seizures in adulthood. This study provides additional evidence for long-term alterations of neuronal excitability and cognitive capacity associated with seizures during development. PMID- 12576175 TI - Intelligence in childhood epilepsy syndromes. AB - Intellectual deficits play a significant role in the psychosocial comorbidity of children with epilepsy. Early educational intervention is critical. OBJECTIVE: This study aims to determine the intellectual ability of children with common childhood epilepsy syndromes-generalised idiopathic epilepsy (GIE), generalised symptomatic epilepsy (GSE), temporal lobe epilepsy (TLE), frontal lobe epilepsy (FLE), central epilepsy (CE) and non-localised partial epilepsy (PE). METHODS: A prospective consecutive series of 169 children were recruited. Epilepsy syndrome was identified by clinical data, seizure semiology, interictal and ictal EEG in each child, using International League Against Epilepsy criteria. Each child had neuropsychology assessment using age-normed and validated instruments. After adjusting for important epilepsy variables, 95% confidence intervals were generated for mean full-scale intelligence quotient (FSIQ) using ANCOVA. RESULTS: Significant differences between epilepsy syndrome groups were found for age of onset (P<0.001), duration of active epilepsy (P=0.027), seizure frequency (P=0.037) and polytherapy (P=0.024). Analysing FSIQ, children with GIE, CE and TLE performed best, and did not differ statistically. Children with GSE had a statistically lower FSIQ than other syndrome groups except PE. FLE functioned significantly better than GSE, but did not differ statistically from other groups. CONCLUSIONS: In childhood epilepsy, delineation of the syndrome has important implications when considering intellectual potential. This information is invaluable in planning educational interventions and supporting the family. PMID- 12576176 TI - Metabolism of brain amino acids following pentylenetetrazole treatment. AB - We studied the effects of pentylenetetrazole (PTZ) on brain amino acid metabolism in mice. Administration of this convulsant did not change forebrain concentrations of amino acids, but when treated animals also received an injection of [15N]leucine, which served as a tracer of brain nitrogen metabolism, total (14N+15N) forebrain [leucine] exceeded control and [glutamate] and [aspartate] were less than control, as were forebrain concentrations of [15N]glutamate and [2-15N]glutamine. These data suggest greater uptake of [15N]leucine but diminished transamination of leucine to glutamate in experimental mice. In contrast to the [15N]leucine studies, which were associated with increased brain [leucine], the administration of [15N]alanine did not alter levels of alanine, glutamate or glutamine. However, label appeared in [2 15N]glutamine much more readily with [15N]alanine than with [15N]leucine as precursor and the ratio of enrichment in [2-15N]glutamine/[15N]alanine was much higher than that in [2-15N]glutamine/[15N]leucine, a finding that is compatible with preferential metabolism of alanine in astrocytes, which are the primary site of brain glutamine synthetase. We conclude that PTZ treatment favors the uptake of selected amino acids such as leucine but also diminishes transamination of leucine to yield glutamate via branched-chain amino acid transaminase. PTZ treatment may favor the "reverse" transamination of 2-keto-isocaproate (KIC), the ketoacid of leucine, to form leucine and to consume glutamate. A net result of these processes may be to enable the brain more readily to dispose of the glutamate that is released from neurons during convulsive activity. PMID- 12576177 TI - Functional and anatomic relationship between cholinergic neurons in the median preoptic nucleus and the supraoptic cells. AB - The median preoptic nucleus (MePO) has been suggested to be an important area in the brain for the regulation of vasopressin (VP) release under the condition of osmotic stimulation. Fos immunoreactivity (Fos-ir), choline acetyltransferase (ChAT) immunoreactivity and retrograde labeling with fluoro-gold were used in this study to determine whether cholinergic neurons in the MePO can be activated by hypertonic NaCl, and to characterize the specific MePO cells that have anatomic projections to the supraoptic nuclei (SON). The results showed that c fos expression specifically induced by hypertonic NaCl was found in the ChAT cells of the MePO. A retrograde tracing experiment demonstrated that the MePO neurons projecting to the SON were cholinergic. In addition, hypertonic saline induced Fos-ir was colocalized with the MePO neurons back labeled with fluoro gold from the SON. Together, these data provide evidence that the MePO cholinergic neurons are activated by osmotic stimulation, and suggest that cholinergic cells in the MePO are functionally important in the control of the SON neurons under the condition of hypertonic stimulation. PMID- 12576178 TI - Nociceptin immunoreactivity and receptor mRNA in the human trigeminal ganglion. AB - Nociceptin is a peptide transmitter belonging to the opioid family. Nociceptin has recently attracted considerable interest since it appears to exhibit a number of differences to the other opioids. In the present study, we used a nociceptin antibody to map the distribution of nociceptin in the human trigeminal ganglion. In addition, we studied the nociceptin receptor at mRNA levels by RT-PCR and the vasomotor response to nociceptin in human cerebral vessels using a sensitive in vitro method. About 70% of all neuronal cells in trigeminal ganglia were nociceptin immunopositive. Nociceptin was predominantly (78%) expressed in medium sized cells (30-60 microm). Nociceptin also distributed in small-sized cells (14% of positive cell bodies; <30 microm) and in large-sized cells (8% of positive cell bodies; >60 microm). Double immunostaining showed that in the human trigeminal ganglion nociceptin colocalized with calcitonin gene-related peptide (CGRP), substance P (SP), nitric oxide synthase (NOS) or pituitary adenylate cyclase activating peptide (PACAP). About 61% of nociceptin positive cells contained CGRP, 54% contained SP, 50% contained NOS and 68% contained PACAP. Immunoreactivity to nociceptin was not detected in human cerebral blood vessels. Reverse transcriptase-polymerase chain reaction detected the expression of nociceptin receptor mRNA in trigeminal ganglia but not in basilar arteries. To further examine whether there are functional nociceptin receptors in human cerebral arteries, a pharmacological study was done, where cerebral arteries revealed strong contractions to 60 mM K(+) and U466166 and strong relaxation to CGRP. Nociceptin failed to elicit contraction or relaxation. In conclusion, nociceptin is expressed in human trigeminal ganglia but not in cerebral blood vessels. Nociceptin is colocalized with CGRP, SP, NOS and PACAP. Nociceptin receptor mRNA is expressed in human trigeminal ganglia but not in basilar arteries. The functional role of nociceptin may be at the presynaptic level. PMID- 12576179 TI - Alterations in hypothalamic-pituitary-adrenal axis activity and in levels of proopiomelanocortin and corticotropin-releasing hormone-receptor 1 mRNAs in the pituitary and hypothalamus of the rat during chronic 'binge' cocaine and withdrawal. AB - Tolerance to the stimulatory effects of cocaine on the hypothalamic-pituitary adrenal (HPA) axis develops after chronic 'binge' cocaine exposure in the rat. This blunting of HPA axis activity in response to cocaine is associated with a cocaine-induced reduction of corticotropin-releasing hormone (CRH) mRNA level in the hypothalamus. There is limited information about the effects of withdrawal from chronic cocaine on HPA activity. The present studies were undertaken to determine levels of the HPA hormones adrenocorticotropic hormone (ACTH) and corticosterone across 10 days of withdrawal following chronic 'binge' pattern cocaine administration (3 x 15 mg/kg/day at hourly intervals) for 14 days. Male Fischer rats showed a significantly attenuated HPA axis response to chronic 'binge' pattern cocaine administration 30 min after the last injection on the 14th day, as measured by both plasma ACTH and corticosterone levels at the nadir time point. Twenty-four hours following the final administration of 'binge' cocaine (the 1st day of withdrawal), a significant elevation of plasma ACTH levels and a modest, but significant, elevation of plasma corticosterone levels were found at the nadir time point. This acute withdrawal-related activation of the hormones of the HPA axis was no longer found on the 10th day of withdrawal. In the anterior pituitary, levels of both proopiomelanocortin (POMC) and CRH receptor 1 (R1) mRNAs were significantly higher than saline controls on the 14th day of chronic 'binge' cocaine and were at control levels on the 4th day of withdrawal. In the neurointermediate lobe of the pituitary, a sustained reduction in POMC mRNA levels was observed on the 3rd, 7th and 14th day of chronic 'binge' cocaine, but POMC mRNA was at control levels by the 4th day of withdrawal. In the hypothalamus, POMC mRNA levels showed a transient decrease on the 1st day of 'binge' cocaine with no change during chronic 'binge' cocaine or its withdrawal. CRH mRNA levels in the hypothalamus were not different from saline controls on the 1st and 4th days of withdrawal. Taken together, the present results show that after development of adaptation or tolerance to chronic 'binge' cocaine there is an increase in HPA activity during acute cocaine withdrawal. In addition to being associated with CRH input from the hypothalamus, the activation of the HPA axis by cocaine withdrawal may be, at least in part, due to the increased POMC and/or CRH-R1 gene expression observed in the anterior pituitary after chronic 'binge' cocaine. PMID- 12576180 TI - Neurodegeneration in autoimmune MRL-lpr mice as revealed by Fluoro Jade B staining. AB - As in many humans suffering from lupus erythematosus, the development of systemic autoimmunity and inflammation in Fas-deficient MRL-lpr mice is accompanied by CNS dysfunction of unknown etiology. Experimental studies revealed infiltration of lymphoid cells into the choroid plexus, reduced neuronal complexity, retarded brain growth, and enlargement of cerebral ventricles. Moreover, an increased presence of cells with nicked-DNA (TUNEL+ cells) in the periventricular areas suggested accelerated apoptosis in brain cells of MRL-lpr mice. However, direct evidence that the dying cells were neurons was lacking. For this purpose, we presently use Fluoro-Jade B (FJB), a novel fluorescent dye which has high affinity for dying neurons (both apoptotic and necrotic). As expected, in comparison to the control groups, the brains of diseased, 5-month-old MRL-lpr mice showed increased numbers of FJB-positive (+) cells in cortical and periventricular regions. The FJB+ cells were significantly more numerous than TUNEL+ cells, and only approximately 7% co-localized with TUNEL. Immunostaining for CD4 and CD8 markers did not correlate with the number of FJB+ cells, suggesting that T-lymphocyte infiltration into the brain tissue is not a reliable predictor of neuronal demise. Conversely, indices of systemic autoimmunity (splenomegaly and high serum anti-nuclear antibody levels) were associated with increased FJB+ cell numbers in brains of autoimmune MRL-lpr mice, supporting the causal link between autoimmunity and neurodegeneration. Taken together, the above results suggest that factors other than T-cell infiltration and cell death mechanisms other than Fas-mediated apoptosis dominate neuronal degeneration in lupus-prone MRL-lpr mice. PMID- 12576181 TI - Neuraminidase activity in different regions of the seizing epileptic and non epileptic brain. AB - The sialic acid in the brain is split from sialoglucoconjugates by sialidases (neuraminidases, EC 3.2.1.18), and is postulated to act as an inhibitor of cellular adhesion and to play a role in various membrane functions. Since epilepsy alters cellular interactions and connectivity, it is reasonable to propose that sialidases can be affected by this pathological state or, alternately, by seizures. Therefore, we studied the activity of total, soluble, and membranal sialidases in various brain regions in normal, kindled epileptic and non-epileptic seizing rats. The results showed that in kindled rats, the total activity of the sialidases significantly decreased in cerebral cortex (11.38%) and cerebellum (28.58%), whereas it increased in brainstem (35.51%), hypothalamus (2.88%) and hippocampus (9.37%). The activity of the membranous sialidases in kindled rats followed the same pattern as the total activity, whereas the activity of soluble sialidase was significantly lower than membranous activity. Interestingly, the activity of total and membranal sialidases in non epileptic seizing rats paralleled that observed in kindled rats. We suggest that the seizure-induced decrease of sialidasic activity may not modify the number of sialic acid molecules bound to gangliosides in cell membranes, as compared to areas of increased activity, that may decrease them. These changes in sialidases' activity may reflect functional disturbances of membrane polysialylated gangliosides related to the functional and anatomical plastic changes associated to seizures. Our data indicate that these changes are related to the presence of seizures rather than to an established epileptic state. PMID- 12576182 TI - Evidence against cholera toxin B subunit as a reliable tracer for sprouting of primary afferents following peripheral nerve injury. AB - In order to investigate whether cholera toxin B subunit (CTb) is transported by unmyelinated primary afferents following nerve injury, we transected the sciatic nerves of six rats, and injected the transected nerves (and in three cases also the intact contralateral nerves) with CTb, 2 weeks later. The relationship between CTb and two neuropeptides, vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY), was then examined in neurons in the ipsilateral L4 and L5 dorsal root ganglia, using immunofluorescence staining and confocal microscopy. We also immunostained sections of spinal cord and caudal medulla for CTb, NPY and VIP. Following nerve section, VIP immunoreactivity was increased in laminae I-II of the spinal cord while NPY immunoreactivity was increased in laminae III-IV of the spinal cord and in the gracile nucleus. On the contralateral side, CTb labelling was detected in laminae I and III-V of the dorsal horn of the L4 and L5 spinal segments, as well as in the gracile nucleus. CTb labelling was seen in the same areas on the lesioned side, but with a dramatic increase in lamina II. No VIP or NPY immunoreactivity was observed in L4 and L5 dorsal root ganglia on the side of the intact nerve, but on the lesioned side VIP was detected in many small neurons and NPY in numerous large neurons. In agreement with the report by Tong et al. [J. Comp. Neurol. 404 (1999) 143], we found that while CTb labelling in the dorsal root ganglion on the side of the intact nerve was mainly in large neurons, on the lesioned side CTb was present in dorsal root ganglion neurons of all sizes. The main finding of the present study was that almost all of the VIP- (96%) and NPY- (98%) positive neurons in the dorsal root ganglia on the lesioned side were also CTb-labelled. After nerve injury VIP is upregulated in fine afferents that terminate in laminae I and II, and most of these probably have unmyelinated axons. Since the cell bodies of these neurons were labelled with CTb that had been injected into the transected sciatic nerve, this suggests that many of these fine afferents, which do not normally transport CTb, are capable of doing so after injury. PMID- 12576183 TI - Effect of proteasome inhibitor on cultured mesencephalic dopaminergic neurons. AB - Proteasomal dysfunction has been implicated in the pathogenesis of Parkinson's disease (PD). We examined the effect of a selective proteasomal inhibitor, epoxomicin, on primary cultured mesencephalic neurons. Exposing rat cultured mesencephalic neurons to epoxomicin for 24 h resulted in neurotoxicity in a dose dependent manner. Epoxomicin caused mitochondrial dysfunction, reduction in reduced glutathione (GSH), and increased generation of free radicals. Neuronal damage was significantly blocked by antioxidative/GSH-augmenting agents. Epoxomicin also increased the expression of Bax and decreased that of Bcl-2, which may cause mitochondrial dysfunction and release of free radicals. Dopaminergic neurons were preferentially resistant to the toxicity of epoxomicin. Inhibiting the synthesis of tetrahydrobiopterin (BH(4)), which has been reported to have antioxidative function, increased the susceptibility of dopaminergic neurons, whereas increasing BH(4) levels protected non-dopaminergic neurons. These findings suggest that BH(4) is at least in part a contributing factor to grand the resistance to dopaminergic neurons against epoxomicin neurotoxicity. Our results suggest that proteasome inhibition causes the neurotoxicity in mesencephalic neurons, but that is not sufficient to reproduce the selective damage to dopaminergic neurons, such as that seen in PD. PMID- 12576184 TI - D2 antagonist-induced c-fos in an identified subpopulation of globus pallidus neurons by a direct intrapallidal action. AB - Much research now supports the view that the dopaminergic innervation of the globus pallidus external segment (GP) influences basal ganglia information processing via pallidal dopamine (DA) D2, D3, and possibly D1 receptors. Systemic DA agonists, or systemic or intrapallidal dopamine D2-class antagonist administration, can induce immediate early gene expression (IEG) in the rat GP. In view of the distinct chemical phenotypes and axonal projections of the GP neurons, it is important to characterize the population(s) of pallidal neurons responding to local DA manipulations. Parvalbumin (PV) immunostaining was used to identify one of the two principal GP neuron populations. Awake, behaving rats received intrapallidal infusions of the dopamine D2 antagonist sulpiride (50 or 100 ng), the D1-class antagonist SCH-23390 (100 ng), the D2-class agonist quinpirole (500 ng), the GABA(A) antagonist picrotoxin (0.25, 0.5 or 1 microg) or bicuculline (20 ng), the GABA(A) agonist muscimol (15 ng) or vehicle. Intrapallidal GABA manipulations were used to assess the likelihood that the effects of the DAergic drugs on Fos induction occurred secondarily to altering intrapallidal GABA release. Using Fos and PV double immunolabeling procedures, we found that several treatments induced GP Fos, but that intrapallidal sulpiride induced Fos almost exclusively in PV-lacking pallidal neurons. No other intrapallidal drug-induced Fos showed similar population specificity. These results support evidence suggesting that GP DA can play a unique and critical role in modulating pallidal neuron function, and that the cessation of pallidal dopamine transmission can activate gene expression within the pallidal neuron subpopulation that maintains extensive axonal projections to caudate-putamen. PMID- 12576185 TI - Estrogen modulation of NMDA-induced seizures in ovariectomized and non ovariectomized rats. AB - The objective of this study was to examine the neuroprotective effects of estrogen in response to N-methyl-D-aspartate (NMDA)-induced seizures in both male and female rats. Thirty-eight Long-Evans rats were divided into five groups: ovariectomized females, non-ovariectomized females, ovariectomized females with estrogen replacement (10 microg 17beta-estradiol in 100 microl sesame oil), males given exogenous estrogen and males receiving no estrogen. Using stereotaxic surgery, a cannula was placed in the lateral ventricle for convulsant agent administration (20 microg of NMDA), while an electrode was placed into the hippocampus for seizure recording. Seizure activity was monitored for 20 min. Onset to first seizure, first seizure duration, seizure frequency and total duration of seizures were determined. Rats were pretreated with either sesame oil (vehicle) or estrogen given subcutaneously for 4 days prior to seizure induction on the fourth day. Rats were euthanized 72 h later and the brains removed for histological processing. Electrode and cannula placement were verified microscopically and neuronal integrity was assessed via hematoxylin and eosin staining. Total seizure number was significantly higher in the ovariectomized females compared to the non-ovariectomized females and the ovariectomized females receiving estrogen (P<0.05). Moreover, hippocampal neuronal damage following seizure induction was significant in the ovariectomized rats compared to the non ovariectomized rats (P<0.05). Pretreatment with estrogen did not affect any of the seizure parameters measured in the male rats. We conclude that estrogen appears to be neuroprotective against NMDA-induced seizures in female ovariectomized rats. PMID- 12576186 TI - Dyrk1A expression pattern supports specific roles of this kinase in the adult central nervous system. AB - Dyrk1A and its Drosophila orthologue, the protein minibrain (mnb), belong to a family of serine/threonine kinases involved in the development of the central nervous system (CNS). However, additional roles for Dyrk1A have to be proposed, as its expression is still prominent in the adult brain. To gain insight into Dyrk1A physiological roles we have studied the distribution of this kinase in the CNS of mice in adulthood. A homogeneous diffuse immunostaining of variable intensity was detected throughout the neuropile, with the white matter displaying faint Dyrk1A immunoreactivity. Dyrk1A immunostaining was strong in the olfactory bulb, the cerebellar cortex and functionally related structures, the spinal cord and most of the motor nuclei of the midbrain and brain stem. These data agree with a possible implication of this kinase in the physiology of olfaction and motor functions. Cellular and subcellular localisation of Dyrk1A was also studied in primary cell culture of cerebellum, one of the structures showing significant Dyrk1A immunostaining in the adult. The distribution of Dyrk1A in primary cell culture showed the presence of this protein in the nucleus and the cytoplasm of both neurons and astrocytes. Moreover, studies on the subcellular distribution of Dyrk1A in whole brain homogenates of adult mice showed the presence of this protein both in nuclear and cytoplasm-enriched fractions, thus supporting selective functions of this kinase in these two subcellular compartments. The present results showing the distribution of Dyrk1A in widespread areas of the adult CNS and in different subcellular compartments, together with previous reports demonstrating its implication in developmental events concur with the idea of several spatio-temporal functional profiles. PMID- 12576187 TI - Colchicine induces apoptosis in organotypic hippocampal slice cultures. AB - The microtubule-disrupting agent colchicine is known to be particular toxic for certain types of neurons, including the granule cells of the dentate gyrus. In this study we investigated whether colchicine could induce such neuron-specific degeneration in developing (1 week in vitro) and mature (3 weeks in vitro) organotypic hippocampal slice cultures and whether the induced cell death was apoptotic and/or necrotic. When applied to 1-week-old cultures for 48 h, colchicine induced primarily apoptotic, but also a minor degree of necrotic cell death in the dentate granule cells, as investigated by cellular uptake of the fluorescent dye propidium iodide (PI), immunostaining for active caspase 3 and c Jun/AP-1 (N) and fragmentation of nuclei as seen in Hoechst 33342 staining. All four markers appeared after 12 h of colchicine exposure. Two of them, active caspase 3 and c-Jun/AP-1 (N) displayed a similar time course and reached a maximum after 24 h of exposure, 24 h ahead of both PI uptake and Hoechst 33342 staining, which together displayed similar time profiles and a close correlation. In 3-week-old cultures, colchicine did not induce apoptotic or necrotic cell death. Attempts to interfere with the colchicine-induced apoptosis in 1-week-old cultures showed that colchicine-induced PI uptake and formation of apoptotic nuclei were temporarily prevented by coapplication of the protein synthesis inhibitor cycloheximide. Application of the pancaspase inhibitor z-VAD-fmk almost completely abolished the formation of active caspase 3 protein and apoptotic nuclei induced by colchicine, but the formation of necrotic nuclei increased correspondingly and the PI uptake was unaffected. We conclude that colchicine induces caspase 3-dependent apoptotic cell death of dentate granule cells in hippocampal brain slice cultures, but the apoptotic cell death is highly dependent on the developmental stage of the cultures. PMID- 12576188 TI - A short half-life GFP mouse model for analysis of suprachiasmatic nucleus organization. AB - Period1 (Per1) is one of several clock genes driving the oscillatory mechanisms that mediate circadian rhythmicity. Per1 mRNA and protein are highly expressed in the suprachiasmatic nuclei, which contain oscillator cells that drive circadian rhythmicity in physiological and behavioral responses. We examined a transgenic mouse in which degradable green fluorescent protein (GFP) is driven by the mPer1 gene promoter. This mouse expresses precise free-running rhythms and characteristic light induced phase shifts. GFP protein (reporting Per1 mRNA) is expressed rhythmically as measured by either fluorescence or immunocytochemistry. In addition the animals show predicted rhythms of Per1 mRNA, PER1 and PER2 proteins. The localization of GFP overlaps with that of Per1 mRNA, PER1 and PER2 proteins. Together, these results suggest that GFP reports rhythmic Per1 expression. A surprising finding is that, at their peak expression time GFP, Per1 mRNA, PER1 and PER2 proteins are absent or not detectable in a subpopulation of SCN cells located in the core region of the nucleus. PMID- 12576189 TI - Age-related microglial activation in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic neurodegeneration in C57BL/6 mice. AB - Microglial activation was investigated in the brains of young (3 months old) and older (9-12 months old) mice following administration of 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP). Tyrosine hydroxylase (TH)-positive neuronal loss differed significantly between young and older mice. Importantly, the two groups clearly demonstrated a distinct microglial activation pattern. In young mice which showed TH neuronal loss at 1 day (33.4%), 3 days (45.1%), 7 days (47.1%) and 14 days (46.9%), microglial activation was first observed at 1 day, with lesser activation at 3 days and none shown later than 7 days. In contrast, in older mice which showed TH neuronal loss at 1 day (49.6%), 3 days (56.1%), 7 days (71.7%) and 14 days (72.1%), microglial activation occurred at 1 day, further intensified at 3-7 days, and was largely abated by 14 days. The double immunohistochemistry further demonstrated that the activated microglia surrounded dopaminergic neurons in older mice at 7 days, which was sharply in contrast to the young mice which were devoid of massive microglial activation in the SN later than 3 days after MPTP treatment. The present study suggests that age-related microglial activation in the SN may be relevant to the higher susceptibility to MPTP neurotoxicity in older mice. PMID- 12576190 TI - Differences in quinpirole-induced local cerebral glucose utilization between naive and sensitized rats. AB - Dopaminergic psychostimulants produce behavioral responses of greater magnitude with repeated, intermittent administration, than a single, acute dose, a phenomenon known as 'sensitization'. Alterations in regional neuronal activity produced by quinpirole, a D(2)/D(3) agonist, in quinpirole-naive and quinpirole sensitized rats were assessed on the basis of local cerebral glucose utilization (LCGU) using the [14C]2-deoxyglucose (2-DG) method. Adult, male Long-Evans rats (180-200 g, n=7-9/group) were subjected to ten injections of quinpirole (0.5 mg/kg, s.c.) administered every 3rd day; controls and quinpirole-naive rats received saline. Locomotor activity was quantitated after injections one and ten to confirm sensitization. The 2-DG procedure was initiated 60 min after an 11th injection in freely moving rats. LCGU was determined in 43 brain regions by quantitative autoradiography. In quinpirole-naive rats, quinpirole decreased LCGU in the caudate/putamen (84% of control), lateral habenula (80% of control), and motor cortex (79% of control). In sensitized rats, quinpirole decreased LCGU in the nucleus accumbens core and shell (77 and 83% of control, respectively) and ventral pallidum (82% of control) as well as in the caudate/putamen (86% of control), lateral habenula (77% of control), and motor cortex (79% of control). This suggests that decreased neuronal activity in the nucleus accumbens and ventral pallidum may underlie the augmented behavioral response to quinpirole in sensitized animals. PMID- 12576191 TI - Naloxone and its quaternary derivative, naloxone methiodide, have differing affinities for mu, delta, and kappa opioid receptors in mouse brain homogenates. AB - Naloxone and naloxone methiodide both act on opioid receptors but naloxone methiodide has limited access to the brain. Naloxone methiodide has been shown to have a lower affinity for opioid receptors than naloxone in the rat and guinea pig but has not been tested in the mouse. We aimed to investigate this by using [3H]DAMGO, [3H]DPDPE and [3H]U-69,593 to compare the ability of naloxone and naloxone methiodide to displace binding to mu, delta and kappa opioid receptors in mouse brain homogenates. Significant binding was observed for each receptor type and the binding affinity for naloxone versus naloxone methiodide was found to be 15:1 for mu, 6:1 for kappa and 330:1 for delta receptors. Therefore, naloxone methiodide does have a lower affinity for opioid receptors than naloxone in mouse brain tissue, which must be taken into consideration in experimental designs. PMID- 12576192 TI - Activation of the hypothalamic-pituitary-adrenal axis following vestibular deafferentation in pigmented guinea pig. AB - Twelve male pigmented guinea pigs underwent either a unilateral vestibular deafferentation (UVD) (n=6) or sham operation (n=6). Compared to the pre-operated salivary cortisol concentrations, the UVD operation resulted in a significant increase in night cortisol concentrations (P<0.05) and a significant interaction between the night cortisol concentration and time (P<0.05). There was no significant difference between the pre- and post-UVD morning salivary cortisol concentrations; nor between the pre- and post-sham morning or night salivary cortisol concentrations. This study suggests that the ocular-motor and postural syndrome is causing the activation of the hypothalamic-pituitary-adrenal (HPA) axis. PMID- 12576193 TI - Nicotine administration decreases neuropeptide Y expression and increases leptin receptor expression in the hypothalamus of food-deprived rats. AB - The effects of nicotine on the expressions of neuropeptide Y (NPY) and leptin receptor in the rat hypothalamus were investigated via immunohistochemistry. The results show that NPY expression is not affected in the arcuate nucleus (ARN) and is increased only slightly in the paraventricular nucleus (PVN) by nicotine administration under normal (i.e. fed) conditions and that leptin receptor expression is decreased slightly in the ARN and not affected in the PVN following nicotine treatment under the same conditions. Food deprivation enhanced NPY and suppressed leptin receptor expression in the ARN and PVN of the hypothalamus. Nicotine administration resulted in decreased NPY and increased leptin receptor levels. PMID- 12576194 TI - Consistent magnetic-field induced dynamical changes in rabbit brain activity detected by recurrence quantification analysis. AB - The reports dealing with the effects of electromagnetic fields (EMFs) on brain electrical activity have been inconsistent. We suspected that the use of linear methods and models accounted for some of the variability, and we explored the issue by using a novel approach to study the effects of EMFs on the electroencephalogram (EEG) from rabbits. The EEG was embedded in phase space and local recurrence plots were calculated and quantified to permit comparisons between exposed and control epochs from individual animals. Statistically significant alterations in brain activity were observed in each animal (n=10) when it was exposed to 2.5 G, 60 Hz, as assessed using each of two recurrence plot quantifiers. Each result was replicated; a positive-control procedure ruled out the possibility that the effect of the field was a product of the method of analysis. Measurements performed while the rabbits were under anesthesia suggested that the effect was mediated by N-methyl-D-aspartate and/or alpha(2) adrenoceptors. No differences were found between exposed and control epochs in any animal when the experiment was repeated after the rabbits had been killed, indicating that a putative interaction between the field and the EEG electrodes could not account for the observed effects. We conclude that EMF transduction resulting in changes in brain electrical activity could be demonstrated consistently using a nonlinear method of analysis. PMID- 12576195 TI - Effect of feeding Murraya koeingii and Brassica juncea diet on [correction] kidney functions and glucose levels in streptozotocin diabetic mice. AB - Purpose of the study was to investigate the effects of daily oral feeding 15% of powdered leaves of Murraya koeingii (MK) (commonly called as Curry patta) and 10% powder of seeds of Brassica juncea (BJ) (commonly called as Rai) for 60 days on serum glucose concentrations and kidney functions in streptozotocin (STZ; 100mg/kg) diabetic rats. Serum glucose levels, body weight, urine volume, serum creatinine, and urinary albumin (UAE) levels were monitored on day 0, 10, 25, 40, and 70 of the experiment. After 60 days of STZ administration, urine volume per day and UAE levels were significantly higher (P<0.0005) in diabetic controls (DC) as compared to normal controls (NC). Although feeding of the MK/BJ showed a trend towards improvement in most of the parameters, results were not statistically different from the DC except in serum creatinine values in BJ-fed rats on day 70. Thus, these plants can be best utilized by promoting them as preferable food adjuvants for diabetic patients. PMID- 12576196 TI - Effect of Nigella sativa on blood hemostatic function in rats. AB - Nigella sativa (NS) is consumed excessively in Saudi Arabia and Gulf Countries but few studies were conducted on its effects on hemostasis. The effects of NS on blood coagulation and some liver function tests of normal adult male albino rats were investigated. Equivalent (180mg NS/kg rat/day), half, double, and triple doses of NS powdered seeds incorporated in a flour dough were administered for 1, 2, and 4 weeks. Controls received plain flour dough. At the end of each feeding period, platelet count, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), levels of fibrinogen, antithrombin III (AT III), and albumin, and activity levels aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were determined. As compared to the control, the equivalent dose of NS induced significant hyperfibrinogenemia (14%) after 4 weeks while the double dose induced significant transient PT prolongation (7.8%) and TT reduction (13%) after 2 weeks and the triple dose induced significant transient APTT reduction (16%), and TT reduction (13%) after 1 week. There was an increase in the albumin level and ALT activity paralleling that of fibrinogen. No changes were noticed in platelet count, AT III level, and AST activity. In conclusion, NS within the doses used seems to induce transient changes in the coagulation activity of rats. PMID- 12576197 TI - Spermicidal activity of the crude ethanol extract of Sedum praealtum in mice. AB - Ethanol extract of Sedum praealtum in intravaginal doses of 10, 20, 40, 50, 100 and 150 mg/kg body weight showed a toxic effect in spermatozoa viability after 24h of administration. Spermatozoa viability was 94, 83, 58 and 24%, respectively, for extract doses of 10, 20, 40 and 50mg/kg body weight, while only dead spermatozoa were found in the vagina of the treated female mice with doses of 100 and 150 mg/kg body weight. PMID- 12576198 TI - Evaluation of the analgesic and antiedematogenic activities of Quassia amara bark extract. AB - We evaluated the possible antiedematogenic, antinociceptive and/or sedative effects of four different extracts obtained from the bark of Quassia amara namely, 70% ethanol (70EtOH), 100% ethanol (100EtOH), dichloromethane (DCM) and hexane extracts (HEX). The oral administration (100, 250 and 500 mg/kg) of these extracts did not show significant effects in any experiment. However, when administered intraperitoneally, the HEX extract decreased the paw edema induced by carrageenan, showed antinociceptive effects on the hot-plate test and on acetic acid-induced writhing, and showed sedative effects on pentobarbital induced sleep. Naloxone did not reverse the antinociceptive effect of this extract. In conclusion, although the mechanisms are uncertain, the results demonstrated that these effects are apparently related to sedative and muscle relaxant or psychomimetic activities of the HEX extract of the plant. PMID- 12576199 TI - A cross-cultural study: anti-inflammatory activity of Australian and Chinese plants. AB - In this study, in vitro inhibitory effects of 33 ethanol extracts obtained from 24 plant species (representing 11 different families) on cyclooxygenase-1 (COX-1) were evaluated. The plant materials selected for this study have been used in aboriginal medicine in Australia and traditional medicine in China for the treatment of various diseases that are considered as inflammation in nature, e.g. asthma, arthritis, rheumatism, fever, edema, infections, snakebite and related inflammatory diseases. All of the selected plants, with one exception, showed inhibitory activity against COX-1, which supports their traditional uses. The most potent COX-1 inhibition were observed from the extracts of Acacia ancistrocarpa leaves (IC(50)=23 microg/ml). Ficus racemosa bark, Clematis pickeringii stem, Acacia adsurgens leaves, Tinospora smilacina stem and Morinda citrifolia fruit powder exhibited inhibition of COX-1 with the IC(50) of 100, 141, 144, 158 and 163 microg/ml, respectively. Aspirin and indomethacin used as the reference COX-1 inhibitors in this study inhibited COX-1 with IC(50) of 241 and 1.2 microg/ml, respectively. The findings of this study may explain at least in part why these plants have been traditionally used for the treatment of inflammatory conditions in Australian aboriginal medicine and traditional Chinese medicine. PMID- 12576200 TI - Modulation of biochemical parameters by Hemidesmus indicus in cumene hydroperoxide-induced murine skin: possible role in protection against free radicals-induced cutaneous oxidative stress and tumor promotion. AB - Hemidesmus indicus has been shown to possess significant activity against immunotoxicity and other pharmacological and physiological disorders. In this communication, we have shown the modulating effect of H. indicus on cumene hydroperoxide-mediated cutaneous oxidative stress and tumor promotion response in murine skin. Cumene hydroperoxide treatment (30 mg per animal) increased cutaneous microsomal lipid peroxidation and induction of xanthine oxidase activity which are accompanied by decrease in the activities of cutaneous antioxidant enzymes and depletion in the level of glutathione. Parallel to these changes a sharp decrease in the activities of phase II metabolizing enzymes was observed. Cumene hydroperoxide treatment also induced the ornithine decarboxylase activity and enhanced the [3H]-thymidine uptake in DNA synthesis in murine skin. Application of ethanolic extract of H. indicus at a dose level of 1.5 and 3.0mg/kg body weight in acetone prior to that of cumene hydroperoxide treatment resulted in significant inhibition of cumene hydroperoxide-induced cutaneous oxidative stress, epidermal ornithine decarboxylase activity and enhanced DNA synthesis in a dose-dependent manner. Enhanced susceptibility of cutaneous microsomal membrane for lipid peroxidation and xanthine oxidase activity were significantly reduced (P<0.01). In addition the depleted level of glutathione, inhibited activities of antioxidants and phase II metabolizing enzymes were recovered to significant level (P<0.05). In summary, our data suggest that H. indicus is an effective chemopreventive agent in skin and capable of ameliorating hydroperoxide-induced cutaneous oxidative stress and tumor promotion. PMID- 12576201 TI - An ethnobotanical study of medicinal plants used by the Zay people in Ethiopia. AB - An ethnobotanical survey was carried out to collect information on the use of medicinal plants by the Zay people who live on islands as well as shore areas of Lake Ziway in the Ethiopian Rift Valley. A total of 33 medicinal plants were reported as being used locally for the treatment and/or control of human and livestock ailments. Results of the survey showed that leaf materials form the major component of plant parts harvested. The majority of the remedies are prepared in the form of juice from freshly collected plant parts. Most of the remedies are prepared from a single species, and are mainly taken orally. Most of the medicinal plants are collected from the wild. Of the total claimed medicinal plants, 10 were reported scarce locally. Environmental degradation and intense deforestation have been reported as the main causes for the depletion of medicinal plants in the area. As the Zay people are still partly dependent on medicinal plants, loss of these plants will, to a certain extent, hamper the existing health care system in the area. Measures for conservation of medicinal plants of the Zay people are urgently needed. PMID- 12576202 TI - Immunomodulatory activity of the aqueous extract from rhizome of Smilax glabra in the later phase of adjuvant-induced arthritis in rats. AB - Our previous paper has reported that the aqueous extract from Rhizoma Smilacis Glabrae (RSG) remarkably inhibited the primary inflammation of adjuvant arthritis (AA) in rats. In the present study, we further examined the activity of RSG and its mechanism on the secondary inflammation of AA. The administration of RSG (400 and 800 mg/kg) during the later phase significantly inhibited the swelling of the adjuvant-non-injected footpad of AA rats. The lipopolysaccharide-induced production of IL-1, TNF and NO by peritoneal macrophages was significantly reduced. In contrast, the extract significantly recovered the decrease in weight gain of the AA rats and Concanavalin A-induced T lymphocyte proliferation and IL 2 production by their splenocytes, while prednisolone (10mg/kg) showed a significant aggravation. Furthermore, RSG significantly recovered the picryl chloride-induced delayed-type hypersensitivity to almost normal levels from the higher or lower levels induced by different treatments of cyclophosphamide with a normalization of CD4/CD8 ratio. These results suggest that RSG exhibit an improvement on AA through down-regulating over-activated macrophages and up regulating the dysfunctional T lymphocytes during the later phase of arthritis. Such characteristics of RSG on AA may be advantageous to the long-term treatment of clinical rheumatoid arthritis. PMID- 12576203 TI - Anti-inflammatory activity of Chinese medicinal vine plants. AB - Anti-inflammatory activities of ethanol extracts from nine vine plants used in traditional Chinese medicine to treat inflammatory conditions were evaluated against a panel of key enzymes relating to inflammation. The enzymes included cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), phospholipase A(2) (PLA(2)), 5-lipoxygenase (5-LO) and 12-lipoxygenase (12-LO). The vine plants studied were: the stem of Spatholobus suberectus Dunn, the stem of Trachelospermum jasminoides Lem., the root from Tripterygium wilfordii Hook. f., the stem of Sinomenium acutum Rehder and Wilson, the stem of Piper kadsura (Choisy) Ohwi, the stem of Polygonum multiflorum Thunb., the root and stem from Tinospora sagittata Gagnep., the root of Tinospora sinensis (Lour.) Merrill, and the stem of Clematis chinensis Osbeck. All of the plant extracts showed inhibitory activities against at least one of the enzymes in various percentages depending upon the concentrations. The extract from S. suberectus was found to be active against all enzymes except COX-2. Its IC(50) values were 158, 54, 31 and 35 microg/ml in COX 1, PLA(2), 5-LO and 12-LO assays, respectively. T. jasminoides showed potent inhibitory activities against both COX-1 (IC(50) 35 microg/ml) and PLA(2) (IC(50) 33 microg/ml). The most potent COX-1, COX-2 and 5-LO inhibition was observed in the extract of T. wilfordii with the IC(50) values of 27, 125 and 22 microg/ml, respectively. The findings of this study may partly explain the use of these vine plants in traditional Chinese medicine for the treatment of inflammatory conditions. PMID- 12576204 TI - Anti-oxidant activities of the extracts from the herbs of Artemisia apiacea. AB - The anti-oxidant activities of the various fractions from the herbs of Artemisia apiacea were investigated. The n-hexane and n-butanol fractions were found to cause significant free radical scavenging effects on DPPH, their scavenging potencies as indicated in IC(50) values, being 230.1 and 183.7 microg/ml, respectively. The n-butanol fraction exhibited a significant decrease in serum transaminase activities elevated by hepatic damage induced by CCl(4)-intoxication in rats. All fractions tested exhibited a lipid peroxidation causing a significant decrease in MDA production in TBA-reactant assay. The n-butanol fraction was the strongest in the increase in the anti-oxidant enzymes such as hepatic cytosolic superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-px) activities in CCl(4)-intoxicated rats. These results suggest that the herbs of A. apiacea possess not only the anti-oxidant, but also the activities in CCl(4)-intoxicated rats. Especially, the n-butanol extract was found to cause significant increases in the rat liver cytosolic SOD, catalase, GSH-px activities as well as a significant decrease in the MDA production. PMID- 12576205 TI - Amaranthus paniculatus (Linn.) improves learning after radiation stress. AB - Brain is highly susceptible to oxidative damage due to its high utilization of oxygen and rather poorly developed antioxidative defense mechanism. Free radicals formation is greatly augmented during ionizing radiation exposure, which causes damage in cerebellum responsible for locomotor activity. Amaranthus paniculatus (Linn.) having high content of beta-carotene (about 15 mg/100g), ascorbic acid, Vitamin C and folate, may prove efficient antioxidants. To evaluate its antioxidative efficacy, healthy Swiss albino mice from an inbred colony were selected and divided into three groups having equal number of male and female in each group. All of these animals were initially trained in Hebb William's Maze, model D(1). After initial training of 10 days, two groups were supplemented with methanolic extract of A. paniculatus (Linn.) at a dose of 600 and 800 mg/kg bw per day, respectively for 15 days. One group without any treatment served as normal. It has been observed that mice, supplemented with extract took lesser time to reach goal than normal (without any treatment). Furthermore after supplementation of Amaranthus, followed by exposure to 9 Gy of gamma radiation by 60Co beam therapy unit, the survived mice took lesser time to reach to their goals than those without plant extract. Control mice (not supplemented with AE extract) showed continuous decline in their learning performance. Mice of Control group died within 12 days after exposure. Irradiated males try to recover from 10th day onwards but they died up to day 12. But in Experimental mice (AE treated), after initial decline in learning ability after exposure, recovery was noticed and not only this 70% of them survived beyond the observation period. Besides male mice showed faster learning ability as compared to females in all groups. After irradiation too, males took lesser time to reach to goals. Learning in all the groups before exposure has been much faster in between 9 and 15 days. After radiation, however it was followed by a sudden spurt and delayed learning response up to 12 days. Recovery was greater in males than females in treated groups. Recovery was greater in males of 600 mg/kg bw per day than other groups. Learning has been almost at the same level from 14th day onward, which indicates that both the dose levels have been found equally effective. PMID- 12576206 TI - Immunobiological features of the galactoside lectin L-Lc isolated from the Argentine mistletoe Ligaria cuneifolia. AB - Ligaria cuneifolia has been used in Argentine folk medicine and is currently employed as substitute for the European mistletoe (Viscum album) as hypotensor agent. Extracts from V. album are widely used in cancer therapy and the antineoplasic effect is attributed to their cytostatic/cytotoxic and immunomodulatory actions. When studying immunomodulatory effects of L. cuneifolia extracts (Lc extracts), they inhibited proliferation of murine mitogen-activated lymphocytes, leukaemic lymphocytes (LB) and breast tumour cells (MMT). The aim of this work was to isolate and identify lectins from Lc extracts and investigate their immunobiological actions. A galactoside lectin (L-Lc) of 57 kDa was isolated. A polyclonal antiserum obtained against Lc extract recognised both L-Lc and MLI (V. album lectin), suggesting the possibility of shared epitopes. Treatment of LB tumour cells with L-Lc (0.01 and 0.1 microg/ml) produced up to 40.0+/-6.9% inhibition of cell growth, which seems partly mediated by apoptosis (apoptosis of L-Lc treated cells 58.4+/-10.3% versus non-treated cells 38.1+/ 8.8%; P<0.05), analysed by acridine orange and ethidium bromide staining. Inhibitory effect on ConA stimulated splenocyte growth was non-significant, while a mitogenic effect was observed on normal murine splenocytes and MMT cells. L-Lc in non-cytotoxic concentrations (250 ng/ml) modified mRNA expression of IL-10 but neither that of TGF-beta nor of IL-2 produced by LB cells. In addition, 43.9+/ 0.5% reduction in NO production by LPS-stimulated murine macrophages was found. Finally, survival rates of LB tumour-bearing mice treated or not with Lc extract or L-Lc failed to show significant differences. PMID- 12576207 TI - IDN 5082, a standardized extract of Salvia miltiorrhiza, delays acquisition of alcohol drinking behavior in rats. AB - Recent research demonstrated that extracts from the dried roots of Salvia miltiorrhiza, a valued folk medicine in China, are effective in reducing voluntary alcohol intake in selectively bred Sardinian alcohol-preferring (sP) rats. These studies were conducted in alcohol-experienced sP rats, i.e. rats which had the opportunity to consume alcohol for several weeks before the test with Salvia miltiorrhiza extracts, reproducing the human "active drinking" phase. In contrast, the present study investigated the effect of IDN 5082, a standardized extract of S. miltiorrhiza, on the acquisition of alcohol drinking behavior in sP rats that had never experienced alcohol before the study. Consistently, the first administration of IDN 5082 (0, 25, 50 and 100mg/kg; i.g.) occurred immediately before alcohol presentation. Alcohol was offered under the two-bottle free-choice regimen with unlimited access for 24h per day. Treatment with IDN 5082 was repeated once daily for 10 consecutive days. IDN 5082 dose dependently delayed acquisition of alcohol drinking behavior; IDN 5082-induced reduction in alcohol intake was compensated by an increase in water intake. These results add further support to the preclinical anti-alcohol profile of S. miltiorrhiza extracts. PMID- 12576208 TI - CNS activity of the methanol extract of Mallotus peltatus (Geist) Muell Arg. leaf: an ethnomedicine of Onge. AB - The aim of the present study was to investigate several neuropharmacological effects of the methanol extract and different fractions of Mallotus peltatus (Geist) Muell Arg. var acuminatus (Euphorbiaceae) leaves in Wistar albino rats and Swiss albino mice. General behavior, exploratory behavior, muscle relaxant activity and phenobarbitone sodium-induced sleeping time were studied. The results revealed that the crude extract at 200-300 mg kg(-1) p.o. and its fractions A and B at 50 mg kg(-1) caused a significant reduction in spontaneous activity (general behavioral profile), remarkable decrease in exploratory behavioral pattern (Y-maze and head dip tests), a reduction in muscle relaxant activity (rotarod, 30 degrees inclined screen and traction tests), and also significantly potentiated phenobarbitone sodium-induced sleeping time. The phytochemical study of crude leaf extract revealed the presence of tannin, triterpenoid, flavonoid, sterol, alkaloid and reducing sugar. Further fractionation and purification yielded two major fractions A (ursolic acid) and B (beta-sitosterol) with some fatty acids as the major compounds. The psychopharmacological activity of the crude leaf extracts appeared to be either due to fraction A (50 mg kg(-1)) or a combination of fractions A and B (50 mg kg( 1)) along with some fatty acids present in the n-butanol part of methanol extract of M. peltatus leaf (MEMPL). PMID- 12576209 TI - Traditional healing with animals (zootherapy): medieval to present-day Levantine practice. AB - Animals and products derived from different organs of their bodies have constituted part of the inventory of medicinal substances used in various cultures since ancient times. This article reviews the history of healing with animals in the Levant (the Land of Israel and parts of present-day Syria, Lebanon, and Jordan, defined by the Muslims in the Middle Ages as Bilad al-Sham) throughout history. Intensive research into the phenomenon of zootherapy in the Levant from early medieval to present-day traditional medicine yielded 99 substances of animal origin which were used medicinally during that long period. Fifty-two animal extracts and products were documented as being used from the early Muslim period (10th century) to the late Ottoman period (19th century). Seventy-seven were recorded as being used in the 20th century. Seven main animal sources have been exploited for medical uses throughout history: honey, wax, adder, beaver testicles, musk oil, coral, and ambergris. The first three are local and relatively easy to obtain; the last four are exotic, therefore, rare and expensive. The use of other materials of animal origin came to an end in the course of history because of change in the moral outlook of modern societies. Among the latter we note mummy, silkworm, stinkbug, scarabees, snail, scorpion, and triton. PMID- 12576210 TI - Traditional medicine in Bulamogi county, Uganda: its practitioners, users and viability. AB - Traditional medicine (TM) in Bulamogi (Uganda) is holistic, providing treatments for physical illnesses as well as psycho-spiritual ones. People use it to prevent and eliminate the effects of witchcraft, to appease spirits and to cure chronic illnesses. The traditional medicine practitioners (TMPs) are numerous and have extensive experience of traditional healing. They serve as important depositories of traditional knowledge of healing. The health seeking behaviour of the Balamogi is biased towards orthodox medicine (OM), because the people believe it to be more effective than TM. Local people prefer the exact diagnosing methods of orthodox medicine practitioners. The functional values of OM and TM are different in that they serve different health needs in the society. We suggest that they are compatible and complementary as a reliance on both systems is observed. The TMPs, upon additional training, are seen by authorities as a ready source of manpower to provide primary health care services. Some effort has been taken to recognise and integrate TM into the mainstream health care delivery system of Uganda. PMID- 12576212 TI - Beneficial effect of aqueous garlic extract on the vascular reactivity of streptozotocin-diabetic rats. AB - The present study evaluated the beneficial effect of aqueous extract of garlic (Allium sativum L.; 100mg/kg/day) on the alterations in vascular reactivity of streptozotocin-diabetic rats. After 8 weeks of treatment, thoracic aortic rings of rats were mounted in organ baths and contractile responses to phenylephrine and relaxant responses to acetylcholine and isosorbide dinitrate were assessed. Induction of diabetes significantly increased contractile responses to phenylephrine and impaired endothelium-dependent relaxations to acetylcholine in aortic rings, but did not change endothelium-independent relaxation to isosorbide dinitrate. Garlic administration significantly improved the impaired endothelium dependent relaxations and decreased the enhanced contractile response to phenylephrine in diabetic rats. It is concluded that intraperitoneal administration of aqueous garlic extract can improve endothelial dysfunction in insulin-dependent model of uncontrolled diabetes. PMID- 12576211 TI - Central nervous system activity of the methanol extract of Ficus platyphylla stem bark. AB - The central nervous system (CNS) activity of the methanolic extract of Ficus platyphylla stem bark was studied on locomotor activity, pentobarbital sleeping time, exploratory behaviour, amphetamine-induced hyperactivity, apomorphine induced stereotypy, active-avoidance and performance on tread mills (rota-rod), using mice and rats. The results revealed that the extract significantly reduced the locomotor and exploratory activities in mice, prolonged pentobarbital sleeping time in rats, attenuated amphetamine-induced hyperactivity and apomorphine-induced stereotypy in mice, dose-dependently. The extract significantly suppressed the active-avoidance response in rats, with no significant effect on motor co-ordination as determined by the performance on rota-rod. The results suggest that the extract may possess sedative principles with potential neuroleptic properties. PMID- 12576213 TI - Lansium domesticum: skin and leaf extracts of this fruit tree interrupt the lifecycle of Plasmodium falciparum, and are active towards a chloroquine resistant strain of the parasite (T9) in vitro. AB - Malaria remains a global problem in the light of chloroquine-resistant strains of Plasmodium falciparum. New compounds are needed for the development of novel antimalarial drugs. Seed, leaf, and fruit skin extracts of Lansium domesticum, a common fruit tree in South-East Asia, are used by indigenous tribes in Sabah, Malaysia for treating malaria. The skin and aqueous leaf extracts of the tree were found to reduce parasite populations of the drug sensitive strain (3D7) and the chloroquine-resistant strain (T9) of P. falciparum equally well. The skin extracts were also found to interrupt the lifecycle of the parasite. The data reported here indicate that extracts of L. domesticum are a potential source for compounds with activity towards chloroquine-resistant strains of P. falciparum. PMID- 12576214 TI - Anti-inflammatory and antipyretic properties of Clerodendrum petasites S. Moore. AB - The methanol extract from Clerodendrum petasites S. Moore (CP extract) was assessed for anti-inflammatory and antipyretic activities on the experimental animal models. It was found that CP extract possessed moderate inhibitory activity on acute phase of inflammation in a dose-related manner as seen in ethyl phenylpropiolate-induced ear edema (ED(50)=2.34 mg/ear) as well as carrageenin induced hind paw edema (ED(30)=420.41 mg/kg) in rats. However, CP extract did not elicit any inhibitory effect on arachidonic acid-induced hind paw edema in rats. In subchronic inflammatory model, CP extract provoked a significant reduction of transudation but had no effect on proliferative phase when tested in cotton pellet-induced granuloma model. CP extract also reduced the alkaline phosphatase activity in serum of rats in this animal model. Moreover, CP extract possessed an excellent antipyretic effect when tested in yeast-induced hyperthermic rats. It is postulated that the anti-inflammatory and antipyretic effects of CP extract are caused by the inhibition of the prostaglandin synthesis. Anyhow, CP extract did not possess any analgesic activity in acetic acid-induced writhing response in mice. The results obtained show that C. petasites has beneficial properties since it possesses potent antipyretic and moderate anti-inflammatory activities without ulcerogenic effect. PMID- 12576215 TI - Daeganghwal-tang inhibits the stem cell factor-induced migration and inflammatory cytokines secretion in mast cells. AB - Traditional Oriental medicinal prescription, Daeganghwal-tang (DGHT) has been used for the treatment of rheumatoid arthritis (RA) in Korea. However, its effect in experimental models remains unknown. Recent reports suggest that in patients with RA, synovial mast cells increase in number and show signs of activation and inflammatory cytokines secretion. Our results show that stem cell factor (SCF) is a potent chemotactic factor for the mast cells in vitro. The chemotactic response to SCF was blocked by DGHT. When DGHT (1mg/ml) was added, the secretion of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6 was inhibited by 60.1, 81.8, 72.5%, respectively in phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187-stimulated HMC-1 cells. In addition, the expression of TNF-alpha mRNA in HMC-1 cells was inhibited by DGHT (1mg/ml). These findings indicate that DGHT inhibits SCF-induced migration and PMA plus calcium ionophore stimulated inflammatory cytokines secretion in mast cells. PMID- 12576216 TI - Inhibitory effect of methanol extract of Euonymus alatus on matrix metalloproteinase-9. AB - Various extracts prepared from stems of Euonymus alatus were tested for cytotoxic activity on human hepatocellular carcinoma cell line, Hep3B cells using the XTT assay method. Also, the extracts were investigated the inhibitory effects on matrix metalloproteinase (MMP)-9 activity using gelatin zymography. The methanol extract, hexane and ethyl acetate fraction exhibited weak cytotoxic activity (IC(50) of >100 microg/ml). However, butanol (IC(50)=65 microg/ml) and chloroform (IC(50)=85 microg/ml) fraction exhibited strongly cytotoxic activity. Gelatin zymography showed that the Hep3B cells secreted matrix metalloproteinase (MMP), probably including MMP-9, which may be involved in tumor cell invasion and metastasis. The butanol fraction showed stronger inhibitory effect of proteolytic activity than other fractions. Also, the butanol fraction was able to decrease the proteolytic activity of MMP-9 in a concentration-dependent manner on zymography. These results suggest that the butanol fraction from E. alatus has highly inhibitory effect on MMP-9 in comparatively low cytotoxicity. PMID- 12576217 TI - Antidiabetic activity of Terminalia pallida fruit in alloxan induced diabetic rats. AB - Different doses of ethanolic fraction of fruits of Terminalia pallida were evaluated for hypoglycemic and antihyperglycemic activity in normal and alloxan diabetic rats. The oral administration of ethanolic extract at a dosage of 0.5 g/kg body weight exhibited a significant antihyperglycemic activity in alloxan diabetic rats, whereas in normal rats no hypoglycemic activity was observed. PMID- 12576219 TI - Lesional expression of a proinflammatory and antiangiogenic cytokine EMAP II confined to endothelium and microglia/macrophages during secondary damage following experimental traumatic brain injury. AB - We analyzed expression of Endothelial Monocyte-Activating Polypeptide II (EMAP II), a proinflammatory, antiangiogenic cytokine in rat brains after stab wound injury and observed a highly significant (p<0.0001) lesional accumulation confined to microglia/macrophages. Maximum numbers were seen at day 5 declining until 21 days after injury. Further, EMAP II(+) microglia/macrophages formed clusters in perivascular Virchow-Robin spaces. Prolonged accumulation of EMAP II(+), ED1(+) microglia/macrophages and increased lesional numbers of EMAP II(+) endothelial/smooth muscle cells during the acute postinjury period might indicate that EMAP II enrich the proinflammatory and antiangiogenic repertoire of effector molecules expressed by activated microglia/macrophages during secondary damage. PMID- 12576220 TI - Lewis hypothalamic cells constitutively and upon stimulation express higher levels of mRNA for pro-inflammatory cytokines and related molecules: comparison with inflammatory resistant Fischer rat hypothalamic cells. AB - Endogenous hypothalamic pro-inflammatory cytokines modulate the hypothalamic pituitary-adrenal (HPA) axis responses. To investigate whether hypothalamic IL 1beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha) are associated with differential inflammatory susceptibilities between Lewis (LEW/N) and Fischer (F344/N) rats, mRNA levels of pro-inflammatory cytokines and related molecules in hypothalamic cell cultures of both strains were quantified by real-time polymerase chain reaction (PCR). In addition to IL-1beta, IL-6, TNF-alpha, and their receptors, LEW/N hypothalamic cells also transcribed more anti-inflammatory molecules, IL-1RII, IL-1RA, and transforming growth factor (TGFbeta1), than F334/N cells. Our findings suggest that a balance exists between transcripts for endogenous pro- and anti-inflammatory molecules in LEW/N rats that may allow them, under basal conditions, to maintain hypothalamic homeostasis and health. However, under stimulated conditions, this balance may be more easily perturbed toward chronic inflammation. PMID- 12576221 TI - Regulatory role of p53 in experimental autoimmune encephalomyelitis. AB - The role of p53, a pro-apoptotic protein, in experimental autoimmune encephalomyelitis (EAE) was investigated using p53-deficient C57BL/6J mice. p53 deficient mice immunised with myelin oligodendrocyte glycoprotein (MOG) exhibited a more severe clinical course of EAE with more severe inflammation in the central nervous system (CNS) compared to wild-type littermates. While T and B cell responses of p53-deficient mice to MOG were comparable to those of wild-type littermates, significantly higher production of IL-6, granulocyte macrophage colony-stimulating factor and IL-10 was observed in lymphocytes exposed to MOG from p53-deficient mice than those from wild-type littermates. Furthermore, a flow cytometric analysis of Annexin V staining showed that apoptosis of CNS infiltrating cells was less in p53-deficient mice with EAE compared to wild-type littermates. These results suggest that p53 may be involved in the regulatory process of EAE through the control of cytokine production and/or the apoptotic elimination of inflammatory cells. PMID- 12576222 TI - Signalling pathways involved in the chemotactic activity of CXCL12 in cultured rat cerebellar neurons and CHP100 neuroepithelioma cells. AB - We compared the signal transduction pathways activated by stromal cell-derived factor-1 (CXCL12) chemokine in two different cell systems: primary cultures of rat cerebellar granule neurons (CGN) and human neuroepithelioma CHP100 cells. Both cell types express functional CXC chemokine receptor 4 (CXCR4), which is coupled both to extracellular signal-regulated kinase (ERK) and Akt phosphorylation pathways. The activation of ERK shows different dependency on the phosphatidylinositol 3-kinase (PI3-K) pathway and different sensitivity to pertussis toxin (PTX) treatment, indicative of coupling to different G proteins in the two cell systems considered. We demonstrate that the inhibition of either the ERK kinase or the PI3-K pathways blocks the CXCL12 induced-chemotaxis in CHP100 cells; while only PI3-K activity is stringently necessary for CGN migration. PMID- 12576223 TI - Catecholamines in a macrophage cell line. AB - This study provides the first evidence for catecholamine synthesis and release in the RAW264.7 cell line, an important macrophage model. Although catecholamines were low in unstimulated cells, activation with lipopolysaccharide (LPS) induced tyrosine hydroxylase (TH) mRNA and increased extracellular norepinephrine and intracellular dopamine within 48 h. The catecholamine synthesis inhibitor alpha methyl-para-tyrosine (alpha-mpt) decreased extracellular norepinephrine levels, suggesting release and rapid turnover of newly synthesized norepinephrine. High concentrations of dopamine or norepinephrine (>/=100 microM) decreased proliferation and increased apoptosis of macrophages. These anti-proliferative effects were prevented by simultaneous treatment with the anti-oxidant ascorbic acid. Pre-incubation with a glutathione synthesis inhibitor (L-buthionine-[S,R] sulfoximine [L-BSO]) increased sensitivity to catecholamine-stimulated apoptosis, suggesting that glutathione protects macrophages from both endogenous and exogenous catecholamines. PMID- 12576224 TI - Experimental autoimmune myasthenia gravis in mice expressing human immunoglobulin loci. AB - Antibodies (Abs) specifically directed against the muscular acetylcholine receptor (AChR) mediate the pathogenesis of myasthenia gravis (MG). The animal model experimental autoimmune MG (EAMG) can be induced by passive transfer or by active immunization of anti-AChR Abs. We report a new EAMG mouse model that generates human anti-AChR Abs upon immunization with Torpedo AChR (tAChR). Mice transgenic for human mu, gamma1, and kappa germ line genes (HuMAb-Mice) were immunized with tAChR. Serum titers of anti-tAChR Abs were in the nanomolar range, and anti-rodent AChR Abs were in picomolar range. Some HuMAb-Mice had signs of muscle weakness, clearly indicating their susceptibility to EAMG. Human Ab-mouse AChR complexes were found at the neuromuscular junction, while AChR loss was up to 65%. Spleen and lymph nodes were used for producing hybridomas. Of the anti tAChR monoclonal Ab-producing hybridomas, 2% had cross-reactivity with rodent AChR and none with human AChR. Immunization with a fusion protein, Trx-Halpha1 210, displaying the human main immunogenic region did not result in EAMG or the generation of human anti-human AChR monoclonal Abs. These experiments show that the HuMAb-Mouse represents a suitable model to generate and study the effects of human anti-AChR Abs in vivo. PMID- 12576225 TI - C1q-calreticulin induced oxidative neurotoxicity: relevance for the neuropathogenesis of Alzheimer's disease. AB - Alzheimer's disease (AD) remains one of the most challenging brain disorders facing modern medicine. Neuronal loss underlies the pathogenesis of AD and can occur, in part, by oxidative stress, by beta-amyloid peptide (Abeta), and by excitotoxins. The complement cascade, especially C1q, may affect reactive oxygen species (ROS) and mediate neuronal injury during AD. We demonstrate that incubation of neurons with purified C1q results in increased ROS, which can be partially blocked by low concentrations of Abeta. C1q-binding sites on neurons were demonstrated by 125I-C1q-binding and immunofluorescence tests performed on primary neurons. The blocking of neuronal calreticulin by its antibody abrogated ROS by C1q. We suggest that the C1q may be an important factor contributing to neuronal oxidative stress and neuronal demise during AD. PMID- 12576226 TI - Vasopressin receptor 1a-mediated negative regulation of B cell receptor signaling. AB - We report here a study of T and B cell development and function in mice with disruption of the vasopressin receptor 1a (v1a) gene. Loss of the v1a receptor caused a shift from IgM(high)/IgD(high) to the more mature IgM(low)/IgD(high) B cells, a significantly greater extent of splenic B cells proliferation in response to anti-IgM stimulation, and enhanced IgG1 and IgG2b production in response to immune challenge with T-dependent antigen. B-1 cells were increased in v1a(-/-) mice. In contrast, T cell differentiation and activation were normal in v1a(-/-) mice. Our data identify a novel function for v1a in the periphery as a negative regulator of B cell receptor (BCR) signaling. These data suggest that in addition to its other stress-related effects, vasopressin may also serve as a counter-regulatory restraint upon the immune system during fight or flight situations. PMID- 12576227 TI - Insulin-like growth factor-I and the cytokines IL-3 and IL-4 promote survival of progenitor myeloid cells by different mechanisms. AB - Hormones, such as insulin-like growth factor-I (IGF-I), and cytokines, like IL-3 and IL-4, promote survival of progenitor myeloid cells. Here we demonstrate that IGF-I, IL-3 and IL-4 all significantly block activation of caspase-3 in promyeloid cells following growth factor deprivation. However, only IL-3 and IGF I increase enzymatic activity and phosphorylation of the survival-promoting kinase Akt. IGF-I fails to reduce caspase-3 activity and cell death in the presence of the PI 3-kinase inhibitors, wortmannin and LY294002, whereas these blockers do not affect the ability of IL-3 to maintain cell survival. IL-4 inhibits caspase-3 activity and promotes promyeloid cell survival by a substrate for PI 3-kinase that is not Akt. These data establish that IGF-I inhibits activation of caspase-3 and promotes promyeloid cell survival through a PI 3 kinase-dependent pathway, whereas IL-3 does not. It therefore appears that signal transduction pathways for all three receptors converge upstream of caspase-3 to prevent apoptosis of progenitor myeloid cells, but their receptors differ in the intracellular substrates that are used to promote cell survival. PMID- 12576228 TI - Intravenous immunoglobulin modulates lymphocyte CD54 and monocyte FcgammaRII expression in patients with chronic inflammatory neuropathies. AB - We studied the expression of different lymphocyte and monocyte cellular determinants involved in leukodiapedesis and antigen presentation in 10 patients with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) and multifocal motor neuropathy (MMN) with persistent conduction blocks before intravenous immunoglobulin (IVIg), immediately after infusion of IVIg and 1 week after infusion. We observed a decrease of T lymphocytes expressing ICAM-1 (CD54) immediately after infusion in 8 out of 10 patients (p<0.04) with a return to pretreatment values after 1 week. The monocytes showed an increase in CD14(+) cells and CD14(+) FcgammaRII inhibitory receptor positive cells, no change in the number of CD14(+) FcgammaRIII activation receptor cells, and an increase in the FcgammaRII/FcgammaRIII ratio on monocytes 1 week after IVIg. Thus, the mechanism of action of IVIg in both CIDP and MMN may involve inhibition of T cell transmigration and modulation of antigen presentation capacities through FcgammaR expression. PMID- 12576229 TI - Human brain endothelial cells supply support for monocyte immunoregulatory functions. AB - Blood-derived monocytic cells comprise a significant component of most inflammatory responses that occur in the CNS. We utilized human brain-derived endothelial cells (HBECs) coated membranes in Boyden chambers to assess immune function related properties of human blood-derived monocytes following interaction with HBECs. Monocytes in contact with HBECs maintained functional antigen-presenting capacity and chemokine/cytokine production in contrast to monocytes that migrated through the HBEC barrier. These results indicate that HBECs, although themselves incapable of serving as competent antigen-presenting cells during the course of inflammatory CNS disorders, supply support needed for infiltrating perivascular monocytes to maintain their functions. Monocyte migration across HBECs was inhibited by interferon-beta. PMID- 12576230 TI - Regulation of prolactin expression in leukemic cell lines and peripheral blood mononuclear cells. AB - To address the role of different intracellular signals in prolactin (PRL) expression in leukocytes, we have investigated the effects of chlorophenylthio cAMP (cptcAMP), phorbol myristate acetate (PMA) and ionomycin on the activation of the upstream PRL promoter in several leukemic cell lines. All three stimulators, alone or in synergism with each other, were able to modulate promoter activity, but their actions were cell-type dependent. In freshly isolated peripheral blood mononuclear cells (PBMC), PRL expression could only be stimulated by cptcAMP. The physiological importance of cAMP in the regulation of PRL expression in leukocytes is suggested by the finding that in PBMC, PRL expression is enhanced by prostaglandin-E(2) and the beta(2)-adrenergic agonist terbutaline, which both signal through cAMP. PMID- 12576231 TI - Anti-myelin oligodendrocyte glycoprotein B-cell responses in multiple sclerosis. AB - Humoral auto-immunity to the myelin oligodendrocyte glycoprotein (MOG) is likely involved in the pathogenesis of multiple sclerosis (MS). In 44 MS patients and 30 controls, Ig-producing B cells were identified by their isotype and as MOG specific spot-forming cells (SFC). Peripheral anti-MOG antibodies were assayed in ELISA as well as anti-butyrophilin antibodies to investigate for molecular mimicry. MS patients had significantly higher levels of IgA- and MOG-SFC than controls, as well as significantly higher antibody responses to MOG and butyrophilin. These data provide added support for the implication of anti-MOG humoral immunity in the pathophysiology of MS, and suggest a balance of systemic (anti-self) and mucosal (environment-modulated) immune reactions in an attempt at regulating the pathogenic specific immune response. PMID- 12576232 TI - Upregulation of the apoptosis regulators cFLIP, CD95 and CD95 ligand in peripheral blood mononuclear cells in relapsing-remitting multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic disease involving an inflammatory reaction within the white matter of the CNS, mediated by T cells, B cells and macrophages. The pathogenesis of MS may involve impaired activation-induced cell death of activated myelin-specific mature T cells. We investigated the mRNA expression of the apoptosis mediators cellular FLICE-inhibitory protein (cFLIP), caspase-8, CD95 and CD95L in peripheral blood mononuclear cells (PB MNCs) from MS patients using real-time PCR. The overall increased expression of the four key players in the CD95 pathway in relapsing-remitting MS suggests their involvement in the inflammatory process in this disease. PMID- 12576233 TI - Interferon-gamma-inducible subunits are incorporated in human brain 20S proteasome. AB - In most tissues expressing MHC class I molecules, proteasomes incorporating IFN gamma-inducible subunits, defined immuno-proteasomes, exist together with constitutive proteasomes. In physiological conditions, the central nervous system expresses neither MHC class I molecules nor TAP1 and TAP2 transporters but besides being constitutive, it is unknown whether immuno-proteasomes are also present in this tissue. We present evidence that in human brain, the two types of proteasome exist suggesting that under physiological conditions, the mechanisms regulating expression of IFN-gamma-inducible subunits as well as of MHC class I molecules and TAP1 and TAP2 transporters in nervous tissue, are not entirely coordinated. PMID- 12576234 TI - NFkappaB and AP-1 DNA binding activity in patients with multiple sclerosis. AB - Current evidence suggests that multiple sclerosis (MS) results from an autoimmune response mediated by T lymphocytes, which would be activated in the peripheral blood and migrate into the central nervous system. NFkappaB and AP-1 are two main transcription factors involved in T-cell activation. To investigate possible alterations in the activity of these factors in MS individuals, we have assayed NFkappaB and AP-1 DNA binding activity in peripheral blood mononuclear cells (PBMC). Binding activity was analyzed by gel mobility shift assay in MS patients compared with controls. No significant differences were found between the two groups, indicating no evidence of abnormalities associated with MS in NFkappaB or AP-1 binding activities in PBMC, both basally and after PMA+anti-CD3 antibody induction. PMID- 12576235 TI - Genetic variations and humoral immune responses to myelin oligodendroglia glycoprotein in adult phenotypes of X-linked adrenoleukodystrophy. AB - The lack of phenotype/genotype association in X-linked adrenoleukodystrophy (X ALD) has prompted the search for disease modifying factors. We previously demonstrated increased serum antibody responses against myelin oligodendrocyte glycoprotein (MOG) in various clinical phenotypes of X-ALD allowing speculations that myelin specific humoral immune responses might be involved in phenotype generation of X-ALD. In the present study, we investigated the possible association of (1) a naturally occurring variable number tandem repeat (vntr) polymorphism (C allele) in the 3' flanking region of the interleukin-6 gene (IL 6), previously demonstrated to modify the course of Alzheimer's disease, systemic lupus erythematodes and Multiple Sclerosis (MS), (2) a tetranucleotide repeat polymorphism (TAAA)(n) in the 3' flanking region of the MOG gene and (3) HLA class II alleles with adult clinical phenotypes and serum antibody responses to MOG in 70 adult X-ALD patients. HLA class II alleles, (TAAA)(n) MOG gene polymorphisms, and IL-6 C allele were not associated with clinical phenotypes. Anti-MOG antibodies were detectable in 17/54 X-ALD patients (31.5%). Anti-MOG antibodies were associated with the 226 bp (TAAA)(n) MOG gene polymorphism but not with distinct clinical phenotypes. PMID- 12576236 TI - Secondary lymphoid organ chemokines are elevated in the cerebrospinal fluid during central nervous system inflammation. AB - Secondary lymphoid organ chemokines have been implicated in chronic inflammation. Their expression in the central nervous system (CNS) has not been studied. Here, levels of secondary lymphoid organ chemokines CCL19 (Exodus-3, MIP-3beta), CCL21 (Exodus-2, 6Ckine, SLC) and CXCL12 (SDF-1alpha) were analysed by ELISA in cerebrospinal fluid (CSF) and plasma from patients with multiple sclerosis (MS); acute optic neuritis (ON) with oligoclonal IgG in the CSF (i.e., first bout of MS); acute ON without oligoclonal IgG (non-MS-type ON); other inflammatory neurological diseases (OIND); and non-inflammatory neurological diseases (NIND). NIND CSF contained CCL19 and CXCL12, while CCL21 was not detected. Intrathecal production of CCL19 and CCL21 was elevated in MS, MS-type ON, and OIND, but not in non-MS-type ON. In MS, CSF levels of CCL19 weakly correlated with CSF cell counts. Intrathecal production of CXCL12 was elevated only in OIND. The role of elevated CCL19 and CCL21 in MS could be retention of mature dendritic cells (DC) in the CNS, recruitment of nai;ve T cells and activated B cells, as well as de novo formation of secondary lymphoid structures in MS plaques. PMID- 12576237 TI - Effects of polyclonal immunoglobulins and other immunomodulatory agents on microglial phagocytosis of apoptotic inflammatory T-cells. AB - T-cell apoptosis in the CNS is an effective mechanism for the noninflammatory resolution of autoimmune T-cell infiltrates. Ingestion of apoptotic leukocytes by microglia results in an efficient clearance of the inflammatory infiltrate, followed by a profound downregulation of proinflammatory phagocyte immune functions. The effects of different immunomodulatory agents on Lewis rat microglial phagocytosis of apoptotic autologous thymocytes or myelin-basic protein (MBP)-specific, encephalitogenic T-cells were investigated using a standardized, light microscopical in vitro phagocytosis assay. Pretreatment of microglia with polyclonal 7S immunoglobulins (IVIg) decreased the phagocytosis of apoptotic thymocytes by 38.2% (p<0.0001). Also, immunoglobulin F(ab')(2) fragments decreased microglial phagocytosis, suggesting an Fc receptor independent mechanism. Similar results were obtained using MBP-specific T-cells. Pretreatment of microglia with IFN-gamma increased the phagocytosis of apoptotic cells by 65.4%, which was to a large extent counteracted by IVIg. Glatiramer acetate (GLAT) did not exert an effect on microglial phagocytosis, while methylprednisolone (MP) induced microglial apoptosis in vitro. These results indicate that IVIg has a high potential to inhibit microglial phagocytosis of apoptotic inflammatory T-cells even under proinflammatory conditions and extend our view of the complex immunomodulatory effects of IVIg. PMID- 12576238 TI - Reactive astrocytes in chronic active lesions of multiple sclerosis express co stimulatory molecules B7-1 and B7-2. AB - Astrocytes in active lesions of multiple sclerosis (MS) express major histocompatibility (MHC) class II molecules, and may play an important role in the presentation of antigen to myelin-specific T cells.However, it has been postulated that astrocytes are unable to act as antigen-presenting cells (APCs) because they would lack the B7 co-stimulatory molecules to activate these T cells. By using double labeling immunofluorescence staining, we demonstrate that reactive astrocytes in chronic active plaques of multiple sclerosis express the co-stimulatory molecules B7-1 and B7-2, and hence have the necessary attributes to act as antigen-presenting cells. PMID- 12576239 TI - Magnesium sulfate for mild preeclampsia. PMID- 12576240 TI - Friedman EA. Primigravid labor: a graphicostatistical analysis. Obstet Gynecol 1955;6:567-89. PMID- 12576241 TI - Magnesium sulfate in women with mild preeclampsia: a randomized controlled trial. AB - OBJECTIVE: To determine whether magnesium sulfate prevents disease progression in women with mild preeclampsia. METHODS: A total of 222 women with mild preeclampsia were randomized to receive intravenous magnesium sulfate (n = 109) or matched placebo (n = 113). Mild preeclampsia was defined as blood pressure of at least 140/90 mm Hg taken on two occasions in the presence of new-onset proteinuria. Patients with chronic hypertension or severe preeclampsia were excluded. Patients were considered to have disease progression if they developed signs or symptoms of severe preeclampsia, eclampsia, or laboratory abnormalities of full or partial HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome. RESULTS: The groups were similar with respect to maternal age, ethnicity, gestational age, parity, and maternal weight at enrollment. Fourteen women (12.8%) in the magnesium group and 19 (16.8%) in the placebo group developed severe preeclampsia after randomization (relative risk = 0.8, 95% confidence interval 0.4, 1.5, P =.41). None in either group developed eclampsia or thrombocytopenia. Women assigned magnesium had similar rates of cesarean delivery (30% versus 25%), chorioamnionitis (3% versus 2.7%), endometritis (5.3% versus 4.3%), and postpartum hemorrhage (1% versus 0.9%), compared to those assigned placebo. Neonates born to women assigned magnesium had similar mean Apgar scores at 1 and 5 minutes as those born to women assigned placebo (7.7 +/- 1.5 versus 7.8 +/- 1.6 and 8.7 +/- 0.7 versus 8.8 +/- 0.6, respectively). CONCLUSION: Magnesium sulfate does not have a major impact on disease progression in women with mild preeclampsia. Magnesium use does not seem to increase rates of cesarean delivery, infectious morbidity, obstetric hemorrhage, or neonatal depression. PMID- 12576242 TI - Seroprevalence of antibodies to Chlamydia pneumoniae in women with preeclampsia. AB - OBJECTIVE: Preeclampsia shares many risk factors and pathophysiologic features with coronary heart disease. We studied whether, like atherosclerosis, preeclampsia is related to seroprevalence of immunoglobulin (Ig) G antibodies to Chlamydia pneumoniae. METHODS: Cross-sectional comparisons were made for 37 women with preeclampsia and 37 women with normal pregnancies at term. In these two groups, antibody titers for IgG, IgM, and IgA seroprevalence to C pneumoniae and IgG to Chlamydia trachomatis and Chlamydia psittaci were compared. RESULTS: Immunoglobulin G antibodies to C pneumoniae at a titer of at least 1:16 were more common in women with preeclampsia (25 of 37) than in women without (15 of 37) (odds ratio 3.1; 95% confidence interval 1.2, 7.9). There were no significant differences in the seroprevalence of IgA or IgM antibodies to C pneumoniae. Women with preeclampsia were also no more likely to have IgG antibodies to C trachomatis or C psittaci. CONCLUSION: Women with preeclampsia had an increased IgG seroprevalence to C pneumoniae but not to C trachomatis or C psittaci. These preliminary data suggest a specific association between infection with C pneumoniae and preeclampsia. PMID- 12576243 TI - Maternal periodontal disease is associated with an increased risk for preeclampsia. AB - OBJECTIVE: To determine if maternal periodontal disease is associated with the development of preeclampsia. METHODS: A cohort of 1,115 healthy pregnant women were enrolled at less than 26 weeks' gestation and followed until delivery. Maternal demographic and medical data were collected. Periodontal examinations were performed at enrollment and within 48 hours of delivery to determine the presence of severe periodontal disease or periodontal disease progression. Preeclampsia was defined as blood pressure greater than 140/90 on two separate occasions, and at least 1+ proteinuria on catheterized urine specimen. The potential effects of maternal age, race, smoking, gestational age at delivery, and insurance status were analyzed, and adjusted odds ratios for preeclampsia were calculated using multivariable logistic regression. RESULTS: During the study period, 763 women delivered live infants and had data available for analysis. Thirty-nine women had preeclampsia. Women were at higher risk for preeclampsia if they had severe periodontal disease at delivery (adjusted odds ratio 2.4, 95% confidence interval 1.1, 5.3), or if they had periodontal disease progression during pregnancy (adjusted odds ratio 2.1, 95% confidence interval 1.0, 4.4). CONCLUSION: After adjusting for other risk factors, active maternal periodontal disease during pregnancy is associated with an increased risk for the development of preeclampsia. PMID- 12576244 TI - Inhibin-A and superimposed preeclampsia in women with chronic hypertension. AB - OBJECTIVE: To determine if maternal serum inhibin-A can be used as a marker for subsequent development of superimposed preeclampsia in women with chronic hypertension. METHODS: Serum for measurement of inhibin-A was obtained at monthly intervals in women with chronic hypertension requiring antihypertensive medications. Superimposed preeclampsia, the primary outcome of interest, was diagnosed when hypertensive women developed proteinuria (at least 300 mg per 24 hour urine specimen). Serum inhibin-A was considered abnormally elevated when the value exceeded the mean plus two standard deviations of the log for chronically hypertensive women who did not develop preeclampsia. RESULTS: A total of 61 women were enrolled in this study, and 21 (34%) developed superimposed preeclampsia. Inhibin-A levels increased with advancing gestational age. Ten women had abnormally increased inhibin-A levels; eight (80%) developed superimposed preeclampsia, compared with 13 of 51 (26%) women with normal inhibin-A levels (P <.001). Sensitivity and specificity were 38% and 95%, respectively, whereas the positive and negative predictive values were 80% and 75%, respectively. CONCLUSION: Although inhibin-A was abnormally increased an average of 3 weeks before the clinical onset of superimposed preeclampsia, the sensitivity of the test as a screen was too limited to be clinically useful. PMID- 12576245 TI - A molecular variant of angiotensinogen is associated with idiopathic intrauterine growth restriction. AB - OBJECTIVE: Intrauterine growth restriction has been associated with failed maternal physiologic changes such as abnormal spiral artery remodeling and reduced maternal blood volume. A polymorphism of angiotensinogen Thr235 has been considered a risk factor for preeclampsia. We genotyped maternal and fetal deoxyribonucleic acid (DNA) for angiotensinogen Thr235 to estimate whether the polymorphism is also a risk factor for intrauterine growth restriction. METHODS: We examined maternal blood DNA in 174 patients with intrauterine growth restriction and 60 patients with both preeclampsia and intrauterine growth restriction. The control group comprised 400 consecutive cases of women with term pregnancies and infants with birth weight between the fifth and 95th percentiles. We also examined 162 DNA samples from fetal blood with intrauterine growth restriction for the Thr235 polymorphism, and 240 normal fetuses were used as the control group. The angiotensinogen genotype was determined using mutagenically separated polymerase chain reaction. The products were size fractionated on an agarose gel. Angiotensinogen genotypes were divided into three groups: MM (homozygous for angiotensinogen Met235 allele), TT (homozygous for angiotensinogen Thr235 allele), and MT (heterozygous). RESULTS: Maternal genotyping revealed a significantly higher Thr235 allele frequency in intrauterine growth restriction (.60) and preeclampsia/intrauterine growth restriction (.63) than in the control group (.36) (P <.001). Fetal genotyping revealed a Thr235 allele frequency of.59 in intrauterine growth restriction fetuses, as compared with the control group (.38) (P <.001). CONCLUSION: Maternal and fetal angiotensinogen Thr235 genotypes are associated with an increased risk of intrauterine growth restriction in our study population. The angiotensinogen Thr235 allele may predispose women to deliver growth-restricted fetuses. PMID- 12576246 TI - Low-dose mifepristone for uterine leiomyomata. AB - OBJECTIVE: To compare the effect of 5 and 10 mg of mifepristone on uterine leiomyoma size and symptoms, and to measure side effects. METHODS: Forty premenopausal women with large, symptomatic leiomyomata were randomized to receive either 5 or 10 mg of mifepristone daily for 6 months in an open-label study. Uterine volume was measured at bimonthly intervals by sonography. Serum concentrations of hemoglobin levels, follicle-stimulating hormone, and liver enzymes were obtained, and endometrial samples, symptoms, and menstrual bleeding were also assessed. RESULTS: Nineteen of 20 subjects taking 5 mg and all 20 subjects taking 10 mg completed all 6 months of the study. Mean uterine volume shrank by 48% (P <.001) in the 5-mg group and 49% (P <.001) in the 10-mg group, a nonsignificant difference. Leiomyoma-related symptoms were comparably reduced in both groups. Amenorrhea occurred in 60-65% of both groups. Hemoglobin levels increased by 2.5 g/dL in anemic subjects. The incidence of hot flashes increased significantly over baseline in the 10-mg group but not in the 5-mg group. Simple endometrial hyperplasia occurred in 28% of all subjects, with no difference between groups. No atypical hyperplasia was noted. CONCLUSION: Mifepristone in doses of 5 mg or 10 mg results in comparable leiomyoma regression, improvement in symptoms, and few side effects. Further study is needed to assess the long-term safety and efficacy of low-dose mifepristone. PMID- 12576247 TI - Reproductive function after conservative surgery and chemotherapy for malignant germ cell tumors of the ovary. AB - OBJECTIVE: To analyze the long-term effects on reproductive function of fertility preserving treatment for malignant germ cell tumors of the ovary. METHODS: A case series analysis was performed on patients with malignant germ cell tumors of the ovary seen or consulted on at our institution between 1975 and 1995. Follow-up information regarding reproductive function was obtained by a mailed or telephone questionnaire. RESULTS: A total of 106 patients with malignant germ cell tumors of the ovary were included in the study. Twenty patients were excluded because of loss of follow-up or death. For the remaining 86 patients, the median follow-up was 122 months (24-384 months). Fertility-preserving surgery was performed in 64 patients. Thirty-eight have attempted conception and 29 have achieved at least one pregnancy (76%). Among the patients who conceived, 20 were International Federation of Gynecology and Obstetrics (FIGO) stage I, one was stage II, and eight were stage III. Sixteen received vincristine, actinomycin D, and cyclophosphamide; three received cisplatin, vinblastine, and bleomycin; three received bleomycin, etoposide, and cisplatin; one received etoposide and cisplatin; four did not receive any chemotherapy; and two were treated with other combinations. Among the nine patients who could not conceive, seven were FIGO stage I and two were stage III. Four of these patients received vincristine, actinomycin D, and cyclophosphamide; three received etoposide and cisplatin; one received cisplatin, vinblastine, and bleomycin; and one patient received no chemotherapy. A total of 38 children were born to these women. Follow-up was available for 16 of these children, who have no evidence of congenital anomalies. CONCLUSION: Fertility-preserving surgery followed by chemotherapy, even in advanced-stage malignant germ cell tumors of the ovary, is effective in conserving the reproductive function of women with malignant germ cell tumors of the ovary. PMID- 12576248 TI - Placental pathology, antiphospholipid antibodies, and pregnancy outcome in recurrent miscarriage patients. AB - OBJECTIVE: To examine whether there are characteristic histological features in placentas from ongoing pregnancies of patients with a history of recurrent miscarriage, with and without primary antiphospholipid antibody syndrome, in relation to clinical pregnancy outcome. METHODS: Patients attending a recurrent miscarriage clinic were investigated and treated according to an established protocol. One hundred twenty-one consecutive patients achieving a potentially viable pregnancy (at least 24 completed weeks' gestation), including 60 primary antiphospholipid antibody syndrome-positive cases and 61 primary antiphospholipid antibody syndrome-negative cases were included. After delivery, placental pathologic examination was carried out by a pathologist unaware of the clinical details. Histological sections were examined by two pathologists independently. Pregnancy outcome and placental findings were reviewed in relation to the maternal antiphospholipid antibody status. RESULTS: Pregnancy outcome was similar in primary antiphospholipid antibody syndrome-positive and primary antiphospholipid antibody syndrome-negative groups regarding gestation at delivery and antepartum obstetric complications. Several histological placental abnormalities were identified in both groups, but most pregnancies were clinically uncomplicated, with no significant placental abnormalities. In cases with pregnancy complications, the placental pathology was primarily that of uteroplacental vasculopathy, such as placental infarction and preeclampsia, but there were no specific placental lesions or patterns of abnormalities characteristic of primary antiphospholipid antibody syndrome-positive patients. A small subgroup of primary antiphospholipid antibody syndrome-positive patients may be at increased risk of development of maternal floor infarction or massive perivillus fibrin deposition. CONCLUSION: There are no specific histopathologic placental abnormalities characteristic of treated patients with antiphospholipid antibody syndrome and poor reproductive history, but complications of uteroplacental disease are more common. PMID- 12576249 TI - Smoking, body mass, and hot flashes in midlife women. AB - OBJECTIVES: To assess whether lifestyle factors, specifically smoking and body mass index (BMI), are associated with the occurrence of any, moderate to severe, or daily hot flashes. METHODS: A cross-sectional study was conducted among women aged 40-60 years residing in the Baltimore metropolitan area who reported their history of hot flashes through a mailed survey. Logistic regression was used to assess the associations between smoking and BMI with any, moderate to severe, and daily hot flashes. RESULTS: Of the 1,087 women included in the study, 56% reported having hot flashes. Compared with never-smokers, current smokers were at an increased risk for both moderate to severe hot flashes (adjusted odds ratio [OR] = 1.9, 95% confidence interval [CI] 1.3, 2.9) and daily hot flashes (adjusted OR = 2.2, 95% CI 1.4, 3.7). Among current smokers, risk for hot flashes increased with greater amount smoked. High BMI (more than 30 kg/m(2)) was associated with an increased risk for moderate to severe hot flashes compared with low BMI (less than 24.9 kg/m(2)) (adjusted OR = 2.1, 95% CI 1.5, 3.0). An increased risk for any or daily hot flashes with high BMI was present only among premenopausal or perimenopausal women. CONCLUSION: Potentially modifiable factors, such as current smoking and high BMI, may predispose a woman to more severe or frequent hot flashes. This information may be valuable for identifying women at risk for hot flashes and for developing appropriate prevention strategies that may include lifestyle modifications. PMID- 12576250 TI - Learning curve in ultrasonographic screening for selected fetal structural anomalies in early pregnancy. AB - OBJECTIVE: To assess the value of first trimester screening by ultrasonography in detecting structural anomalies of the fetus in a general obstetric population. METHODS: During 1993-1998, 20,465 consecutive pregnant women who resided in a defined geographic area participated in ultrasonographic screening for major malformations. These included anomalies of the central nervous system, urinary tract, abdominal wall, and long bones. Heart anomalies were not expected to be detected. The examinations were offered at 13-14 weeks' gestation as part of routine maternal care and were done by specially trained midwives. The pregnancy outcomes were ascertained from obstetric and pediatric records, and the data were completed by information from the national birth and malformation registries. RESULTS: A total of 307 fetuses (1.5%) with a major malformation were found; 67 fetuses (0.3%) had noncardiac major structural defects expected to be detectable by ultrasonography in early pregnancy. Thirty-five of 67 (52%) were identified at the early scan. Sensitivity for these defects increased from 22% to 79% from the first to the last (sixth) study year (P =.009). CONCLUSION: In a low-risk population, adequate sensitivity in screening for major malformations by early ultrasonography can be achieved after a learning curve of 3-4 years. PMID- 12576251 TI - Incidence of postpartum lumbosacral spine and lower extremity nerve injuries. AB - OBJECTIVE: Neurological injury associated with present day labor and delivery is thought to be unusual. The purpose of this study was to estimate the incidence, severity, and duration of postpartum lumbosacral spine and lower extremity nerve injury and identify factors related to nerve injury. METHODS: All women who delivered a live-born infant from July 1997 through June 1998 were asked about symptoms of lumbosacral spine and lower extremity nerve injury the day after delivery. Women with symptoms were examined by a physiatrist to confirm injury, and their cases were then followed by telephone until the symptoms resolved. Maternal variables (including prospective documentation of time spent pushing in various positions) and fetal variables that might be associated with risk of nerve injury were compared between women with injury and those without. RESULTS: Six thousand fifty-seven women delivered live-born infants; 6,048 were interviewed and 56 had a confirmed new nerve injury, an incidence of 0.92%. Factors found by logistic regression analysis to be associated with nerve injury were nulliparity and prolonged second stage of labor. Women with nerve injury spent more time pushing in the semi-Fowler-lithotomy position than women without injury. The median duration of symptoms was 2 months. CONCLUSION: The estimated incidence of postpartum nerve injury was greater than reported from previous studies and is associated with nulliparity and prolonged second stage of labor. PMID- 12576252 TI - Pregnancy-related mortality in the United States, 1991-1997. AB - OBJECTIVE: To describe trends in pregnancy-related mortality and risk factors for pregnancy-related deaths in the United States for the years 1991 through 1997. METHODS: In collaboration with the American College of Obstetricians and Gynecologists and state health departments, the Pregnancy Mortality Surveillance System, part of the Division of Reproductive Health at the Centers for Disease Control and Prevention, has collected information on all reported pregnancy related deaths occurring since 1979. Data include those present on death certificates and, when available, matching birth or fetal death certificates. Data are reviewed and coded by clinically experienced epidemiologists. The pregnancy-related mortality ratio was defined as pregnancy-related deaths per 100,000 live births. RESULTS: The reported pregnancy-related mortality ratio increased from 10.3 in 1991 to 12.9 in 1997. An increased risk of pregnancy related death was found for black women, older women, and women with no prenatal care. The leading causes of death were embolism, hemorrhage, and other medical conditions, although the percent of all pregnancy-related deaths caused by hemorrhage declined from 28% in the early 1980s to 18% in the current study period. CONCLUSION: The reported pregnancy-related mortality ratio has increased, probably because of improved identification of pregnancy-related deaths. Black women continue to have an almost four-fold increased risk of pregnancy-related death, the greatest disparity among the maternal and child health indicators. Although review of pregnancy-related deaths by states remains an important public health function, such work must be expanded to identify factors that influence the survival of women with serious pregnancy complications. PMID- 12576253 TI - Application of electric field stimulation for investigations of human placental blood vessels. AB - OBJECTIVE: To test electric field stimulation on human placental vessels. METHODS: The effects of electric field stimulation on placental vessels were examined in an isometric myograph. RESULTS: Electric field stimulation induced contractions in human placental blood vessels in vitro under isometric conditions when bubbling carbogen through the organ bath. After reaching half-maximal contractions, the vessel rings showed spontaneous relaxation. Pretreatment with verapamil (10(-6) mol/L) or nickel (Ni(2+)) (2 mmol/L) inhibited the contractions to a magnitude of 63.81% +/- 7.69% and 88.36% +/- 12.17% (mean +/- standard error of the mean), respectively. In calcium (Ca(2+))-free medium after combined cyclopiazonic acid (10(-5) mol/L) and Ni(2+) treatment, it was not possible to elicit contractions with electric field stimulation. Bubbling through physiologic in utero hypoxic gases enhanced the contractile responses of the human placental vessel rings to electric field stimulation. The spontaneous relaxation of the veins was not altered, but those of the arteries were reduced to zero. Testing the same gases on mesenteric arteries of rats had an opposite effect concerning contractility. Sodium nitrite decreased the contractions of the placental vessel rings, but the efficacy was decreased by the in utero gases. CONCLUSION: Electric field stimulation has a direct, non-neurogenic contractile effect on isolated placental vessels, which mainly depends on the influx of extracellular Ca(2+) and on a mechanism independent of intracellular Ca(2+) concentration elevation. Physiologic hypoxia has a stimulatory effect on the contractility of human placental vessels, therefore in utero gases should be used instead of carbogen gas; and electric field stimulation is a suitable method for the investigation of the direct effects of pharmacologic agents on human placental vessels. PMID- 12576254 TI - Does cesarean delivery prevent anal incontinence? AB - OBJECTIVE: To compare the incidence and severity of anal incontinence in primiparas after cesarean delivery versus spontaneous vaginal delivery. METHODS: A total of 184 primiparas who delivered by cesarean (104 emergency, 80 elective) and 100 who delivered vaginally were interviewed 10 +/- 2 months postpartum. A comprehensive bowel function questionnaire was completed. Bowel-specific questions included bowel habits, laxative use, urgency, flatus, urge and passive incontinence, soiling, and pad use, before and during pregnancy and postpartum. Obstetric details were confirmed from obstetric records. RESULTS: Anal incontinence was first present in nine (5%) mothers after cesarean delivery and eight (8%) after vaginal delivery (relative risk 0.611, 95% confidence interval 0.25, 1.53). Severe symptoms necessitating pad use affected two (3%) mothers after elective cesarean and one (1%) after vaginal delivery. Two (3%) mothers after elective cesarean, one (1%) after emergency cesarean, and two (2%) after vaginal delivery had at least two symptoms. Anal incontinence followed prelabor emergency cesarean in two mothers. Of the 22 mothers who sustained a second degree tear, five (23%) had new anal incontinence compared with only one (3%) of 40 mothers with an intact perineum (Fisher exact test value = 9.697, P =.014). CONCLUSION: Because severe anal incontinence followed elective and prelabor emergency cesarean, it seems that pregnancy itself can lead to pelvic floor disorders. A high incidence of anal incontinence is associated with a second degree tear. Measures to detect and reduce postpartum anal incontinence should target all pregnant women and mothers, even after prelabor cesarean delivery. PMID- 12576255 TI - Pelvic floor muscle training during pregnancy to prevent urinary incontinence: a single-blind randomized controlled trial. AB - OBJECTIVE: Urinary incontinence is a chronic health complaint that severely reduces quality of life. Pregnancy and vaginal delivery are main risk factors in the development of urinary incontinence. The aim of this study was to assess whether intensive pelvic floor muscle training during pregnancy could prevent urinary incontinence. METHODS: We conducted a single-blind randomized controlled trial at Trondheim University Hospital and three outpatient physiotherapy clinics in a primary care setting. Three hundred one healthy nulliparous women were randomly allocated to a training (n = 148) or a control group (n = 153). The training group attended a 12-week intensive pelvic floor muscle training program during pregnancy, supervised by physiotherapists. The control group received the customary information. The primary outcome measure was self-reported symptoms of urinary incontinence. The secondary outcome measure was pelvic floor muscle strength. RESULTS: At follow-up, significantly fewer women in the training group reported urinary incontinence: 48 of 148 (32%) versus 74 of 153 (48%) at 36 weeks' pregnancy (P =.007) and 29 of 148 (20%) versus 49 of 153 (32%) 3 months after delivery (P =.018). According to numbers needed to treat, intensive pelvic floor muscle training during pregnancy prevented urinary incontinence in about one in six women during pregnancy and one in eight women after delivery. Pelvic floor muscle strength was significantly higher in the training group at 36 weeks' pregnancy (P =.008) and 3 months after delivery (P =.048). CONCLUSION: Intensive pelvic floor muscle training during pregnancy prevents urinary incontinence during pregnancy and after delivery. Pelvic floor muscle strength improved significantly after intensive pelvic floor muscle training. PMID- 12576256 TI - Evaluation of the fascial technique for surgical repair of isolated posterior vaginal wall prolapse. AB - OBJECTIVE: To study the anatomic and functional efficacy and assess long-term success of the fascial technique in the repair of rectocele. METHODS: Forty-two women with symptomatic posterior vaginal wall prolapse of at least stage II underwent a surgical repair using the technique of reconstruction of the rectovaginal septum. These women were evaluated at 6 weeks and 18 months postoperatively for anatomic improvement in the grade of their rectocele and a functional improvement in their vaginal, bowel, and sexual symptoms. RESULTS: Ninety-five percent (40 of 42) were assessed at 6 weeks and 78.5% (33 of 42) attended follow-up at 18 months. Preoperative symptoms included 1) vaginal protrusion (78%); 2) defecation symptoms (76%), which included fecal incontinence alone in 9.5%, evacuation difficulties in 57%, and both fecal incontinence and evacuation difficulties in 9.5%; and 3) sexual dysfunction (33%). At 6-week follow-up there was resolution of vaginal protrusion in 87.5%, and bowel symptoms in 87%. At 18 months there was anatomic cure in 92%, improvement in defecation in 81%, and improvement of sexual dysfunction in 35%. No major complications were seen. CONCLUSION: This technique is effective in providing relatively long anatomic cure of the rectocele and resolution of its symptoms. PMID- 12576257 TI - Selecting suspension points and excising the vagina during Michigan four-wall sacrospinous suspension. AB - OBJECTIVE: To describe the variations in the location of the vaginal apex and the length of vagina excised in women undergoing the Michigan four-wall sacrospinous suspension for posthysterectomy vaginal vault prolapse. METHODS: A prospective observational study of 76 women who had the Michigan modification sacrospinous suspension performed between 1998 and 2001 for posthysterectomy vaginal vault prolapse was carried out. Demographics and preoperative, operative, and postoperative findings were noted, including the pelvic organ prolapse quantification score. The locations of the suspension points relative to the hysterectomy scar were recorded. The amount of vagina excised at surgery and the pre- and postoperative vaginal lengths are reported. RESULTS: The mean length and standard deviation of vagina excised was 4.6 +/- 2.5 cm. The apex created at sacrospinous fixation was at the hysterectomy scar in only seven women (9%). It was most often situated behind the hysterectomy scar, in 58 cases (76%); it was situated in front of it in 11 (14%). In seven women no vagina was excised, and in the remaining 69 women a mean length of 5.1 +/- 2.2 cm was removed. The mean vaginal lengths were 9.7 +/- 1.7 cm preoperatively and 9.4 cm +/- 0.8 postoperatively, a 0.3-cm difference. CONCLUSION: When one performs the Michigan modification sacrospinous suspension, the chosen suspension points are often not at the hysterectomy scar, and in women with large prolapses excess vagina frequently is excised without compromising postoperative vaginal length. PMID- 12576258 TI - Characterization of the relationship between joint laxity and maternal hormones in pregnancy. AB - OBJECTIVE: To evaluate peripheral joint laxity during pregnancy and to correlate changes with serum cortisol, estradiol, progesterone, and relaxin. METHODS: Forty six women with first-trimester singleton gestations consented to participate in this longitudinal observational study. Bilateral wrist laxity measurements (flexion-extension and medial-lateral deviation) were made using a clinical goniometer, and serum levels of cortisol, estradiol, progesterone, and relaxin were determined during each trimester of pregnancy and postpartum. Patients were also screened for subjective joint complaints. Statistical analysis included Student t test, analysis of variance, and linear regression analysis. RESULTS: Eleven women (24%) were excluded from the study after spontaneous first-trimester pregnancy loss. Fifty-four percent (19 of 35) demonstrated increased laxity (10% or higher) in either wrist from the first to the third trimester. Although serum levels of cortisol, estradiol, progesterone, and relaxin were significantly elevated during pregnancy, no significant differences in these levels were noted between those who became lax during gestation and those who did not. Linear regression analysis of wrist joint laxity and level of serum estradiol, progesterone, and relaxin demonstrated no significant correlation. Wrist flexion extension laxity, however, did significantly correlate with level of maternal cortisol (r = 0.18, P =.03). Fifty-seven percent of women developed subjective joint pain during pregnancy, which was not associated with increased joint laxity, but was associated with significantly increased levels of estradiol and progesterone. CONCLUSION: Peripheral joint laxity increases during pregnancy; however, these changes do not correlate well with maternal estradiol, progesterone, or relaxin levels. PMID- 12576260 TI - Effect of raloxifene on the response to conjugated estrogen vaginal cream or nonhormonal moisturizers in postmenopausal vaginal atrophy. AB - OBJECTIVE: To study the effect of raloxifene on the response to conjugated estrogen cream or nonhormonal moisturizer in postmenopausal women with preexisting signs of vaginal atrophy. METHODS: Postmenopausal women with preexisting and untreated vaginal atrophy were enrolled in this parallel, placebo controlled, randomized study. A total of 187 women were randomized to four treatment groups: daily oral raloxifene (60 mg per day) or a placebo in a double blind manner plus one application of conjugated estrogen cream (0.5 g) or one applicator full of nonhormonal moisturizer, open label. The conjugated estrogen cream or nonhormonal moisturizer was applied daily for the first 2 weeks, and then twice weekly thereafter for 3 months. Efficacy of treatment regimens on signs and symptoms of vaginal atrophy was evaluated by monitoring objective and subjective parameters. RESULTS: Signs and symptoms of vaginal atrophy improved in all four treatment groups. Raloxifene did not diminish the magnitude of improvement when administered with either vaginal preparation. Conjugated estrogen cream produced a statistically greater improvement in signs (P <.05) but not in individual symptoms or overall satisfaction relative to nonhormonal moisturizer. CONCLUSION: Postmenopausal women with evidence of preexisting vaginal atrophy may use either low-dose conjugated estrogen cream or nonhormonal moisturizer to treat the atrophy concurrently with raloxifene (60 mg per day). PMID- 12576259 TI - Gabapentin's effects on hot flashes in postmenopausal women: a randomized controlled trial. AB - OBJECTIVE: To evaluate whether treatment with the anticonvulsant gabapentin may be effective in reducing hot flash frequency and severity. METHODS: A randomized, double-blind, placebo-controlled trial was conducted in 59 postmenopausal women with seven or more hot flashes per day examining the effects of gabapentin 900 mg per day on hot flash frequency after 12 weeks of treatment. Subsequently, study patients were enrolled in a 5-week, open-label treatment phase, during which patients could increase the dose of gabapentin to 2,700 mg per day, if needed. RESULTS: After 12 weeks of double-blind treatment, intention-to-treat analysis showed that gabapentin 900 mg per day was associated with a 45% reduction in hot flash frequency and a 54% reduction in hot flash composite score (frequency and severity combined into one score) from baseline, compared with 29% (P =.02) and 31% (P =.01) reductions, respectively, for placebo. Four patients (13%) in the gabapentin group and one (3%) in the placebo group withdrew from the double-blind study because of adverse events. Fifteen patients (50.0%) in the gabapentin group reported at least one adverse event, compared with eight patients (27.6%) in the placebo group. Higher, open-label gabapentin dosing was associated with 54% and 67% reductions in hot flash frequency and composite score from baseline, respectively. CONCLUSION: Gabapentin is effective in reducing hot flash frequency and severity in postmenopausal women. PMID- 12576261 TI - Effect of raloxifene on sexual function in older postmenopausal women with osteoporosis. AB - OBJECTIVE: To assess the effect of raloxifene compared with placebo on sexual function in older postmenopausal women undergoing therapy for the treatment of osteoporosis. METHODS: A subset (12%) of English-speaking women in the United States and Canada participating in the Multiple Outcomes of Raloxifene Evaluation Trial were asked to complete a sexual function questionnaire at baseline and after 36 months of treatment. The Multiple Outcomes of Raloxifene Evaluation Trial is a multicenter, randomized, blinded, placebo-controlled clinical trial, in which 7,705 postmenopausal women with osteoporosis were randomly assigned to one of three groups: raloxifene hydrochloride 60 mg per day or 120 mg per day or placebo. In this substudy, 943 women completed the sexual function questionnaire at both visits. Because preliminary analyses showed no differences by raloxifene dose (n = 302 for 60 mg per day; n = 322 for 120 mg/day), the two groups were combined and compared with the placebo group (n = 319). For the given sample size, we had 80% power (alpha =.05, two-sided, ratio of raloxifene to placebo = 2:1) to detect a 10%-16% difference in the proportion of women experiencing no change in sexual function between placebo and treatment groups. RESULTS: Overall, sexual function and changes in sexual function from baseline to study end between the raloxifene and placebo groups did not differ. In particular, there were no differences in sexual desire or frequency of sexual activity between the groups. Among sexually active women, there were no differences in enjoyment, satisfaction, orgasm, or reported sexual problems. CONCLUSION: Sexual function in older postmenopausal women with osteoporosis is not affected by treatment with raloxifene. PMID- 12576262 TI - Effects of phenobarbital and multiple-dose corticosteroids on developmental outcome at age 7 years. AB - OBJECTIVE: To determine the effects of phenobarbital and repeated antenatal steroid use on the primary outcome (intelligence and achievement) and secondary outcomes (behavior and head circumference) at age 7 years. METHODS: This study was a secondary analysis of a double-blind clinical trial (phenobarbital-vitamin K versus placebo). Intelligence (Wechsler Intelligence Scale for Children, third edition), achievement (Wide Range Achievement Test, third edition), and behavioral (Achenbach Teacher's Report Form and the Child Behavioral Check List) testing and head circumference measurement were performed on 7-year-old children whose mothers participated in a trial to determine if antenatal phenobarbital and vitamin K prevented severe intracranial hemorrhage. Antenatal steroid therapy in these women was repeated weekly. RESULTS: Two hundred ninety-nine of 372 newborns (80%) whose mothers participated in the trial were followed up at age 7 years. Comparing mean (+/- standard deviation [SD]) Wechsler Intelligence Scale for Children scores in the placebo versus treatment groups, there were no differences (P >.4) in Full Scale IQ (100.3 +/- 14.2 versus 100.6 +/- 14.2), Performance IQ (100.3 +/- 14.6 versus 101.5 +/- 15.6), and Verbal IQ (100.2 +/- 14.9 versus 99.6 +/- 13.7). Comparing mean (+/- SD) Wide Range Achievement Test standardized scores in the placebo versus treatment groups, there were no differences (P >.4) in Reading (97.3 +/- 13.9 versus 98.0 +/- 14.9), Spelling (95.8 +/- 12.7 versus 95.3 +/- 13.3), Mathematics (95.9 +/- 13.8 versus 94.5 +/- 14.5), or head circumference measurements (20.3 +/- 0.6 cm versus 20.4 +/- 0.6, P >.2). Similarly, there were no differences in the mean Achenbach Teacher's Report Form and Child Behavioral Check List scores between the placebo and treatment groups. CONCLUSION: Antenatal phenobarbital and repetitive antenatal corticosteroid therapy was not associated with adverse effects on intelligence, achievement, behavior, or head circumference at age 7 years. PMID- 12576263 TI - Pregnancy-related substance use in the United States during 1996-1998. AB - OBJECTIVE: To provide a baseline estimate of the national prevalence of pregnancy related illicit drug use and abstinence rates. METHODS: We analyzed data collected between 1996 and 1998 from the National Household Survey on Drug Abuse, a nationally representative sample survey of 22,303 noninstitutionalized women aged 18-44 years, of whom 1,249 were pregnant. RESULTS: During 1996-1998, 6.4% of nonpregnant women of childbearing age and 2.8% of pregnant women reported that they used illicit drugs. Of the women who used drugs, the relative proportion of women who abstained from illicit drugs after recognition of pregnancy increased from 28% during the first trimester of pregnancy to 93% by the third trimester. However, because of postpregnancy relapse, the net pregnancy-related reduction in illicit drug use at postpartum was only 24%. Marijuana accounted for three fourths of illicit drug use, and cocaine accounted for one-tenth of illicit drug use. Of those who used illicit drugs, over half of pregnant and two-thirds of nonpregnant women also used cigarettes and alcohol. Among the sociodemographic subgroups, pregnant and nonpregnant women who were young (18-30 years) or unmarried, and pregnant women with less than high school education had the highest rates of illicit drug use. CONCLUSION: The continued burden of illicit drug use during pregnancy calls for policy efforts to enable primary care providers to identify and refer women who use substances to treatment and support services. Prevention of uptake of illicit drug use should be an integral part of public health programs for young women. PMID- 12576264 TI - Screening for gestational diabetes: a summary of the evidence for the U.S. Preventive Services Task Force. AB - OBJECTIVE: To systematically review the evidence for screening for gestational diabetes mellitus (GDM). DATA SOURCES: We established eligibility criteria for relevant studies. We systematically searched MEDLINE and the Cochrane Collaboration Library for studies meeting eligibility criteria. We supplemented this search with further studies identified from reference lists of reviews. METHODS OF STUDY SELECTION: Two reviewers examined each article for eligibility. A single reviewer abstracted relevant data from the included articles; a second reviewer checked the abstractions. We graded the quality of the articles according to criteria developed by the U.S. Preventive Services Task Force. TABULATION, INTEGRATION, AND RESULTS: No well-conducted, randomized, controlled trial provides direct evidence for the health benefits of screening for GDM. The evidence is unclear regarding the optimal screening and reference diagnostic test for GDM. The impact of hyperglycemia on adverse maternal and neonatal health outcomes is probably continuous. Although insulin therapy decreases the incidence of fetal macrosomia for those women with more severe degrees of hyperglycemia, the magnitude of any effect on maternal and neonatal health outcomes is not clear. The evidence is insufficient to determine the magnitude of health benefit for any treatment among the large number of women with GDM at milder degrees of hyperglycemia. We found limited evidence regarding the potential adverse effects of screening for GDM. CONCLUSION: Because of the lack of high-quality evidence concerning critical issues, we are unable to determine the extent to which screening has an important impact on maternal and neonatal health outcomes. A randomized, controlled trial of screening is necessary to answer the many remaining questions. PMID- 12576265 TI - Screening for gestational diabetes mellitus: recommendations and rationale. PMID- 12576266 TI - A Darwinian view of obstructed labor. AB - This essay discusses the evolutionary biology of dystocia. From a Darwinian standpoint, the high frequency of dystocia observed today seems evolutionarily untenable. Hunter-gatherers, most notably the Inuit, appear not to suffer from dystocia. It may be that people from an agriculture-based background are, obstetrically speaking, less well adapted to the good nutrition of a modern affluent diet. PMID- 12576267 TI - Prediction and early detection of preterm labor. AB - Studies of cervical sonography, fetal fibronectin, and uterine contraction monitoring during pregnancy have improved our understanding of how preterm labor occurs, but their use in practice remains uncertain. This article reviews the use of these tests to improve the accuracy of diagnosis of preterm labor and to estimate the likelihood of preterm birth in women with historical risk factors. Because the clinical criteria for a diagnosis of preterm labor are inaccurate until labor is well established, over-diagnosis is common. A cervical length measurement of more than 30 mm or a negative fibronectin obtained from a patient with possible preterm labor can avoid over-diagnosis and unnecessary treatment. There is no role for routine use of either cervical sonography or fibronectin to screen pregnant women for preterm birth risk, but women thought to be at increased risk may be reassured by negative test results. Uterine contraction monitoring has low sensitivity for detecting women at risk of preterm birth. Current use of cervical length and fetal fibronectin in pregnancy is limited to situations where a negative result can avoid unnecessary interventions. PMID- 12576268 TI - Vaginal uterine artery ligation avoids high blood loss and puerperal hysterectomy in postpartum hemorrhage. PMID- 12576269 TI - Vaginal uterine artery ligation avoids high blood loss and puerperal hysterectomy in postpartum hemorrhage. PMID- 12576270 TI - Benefits of term delivery in infants with antenatally diagnosed gastroschisis. PMID- 12576272 TI - Dipeptides containing the alpha-aminoisobutyric residue (Aib) as ligands: preparation, spectroscopic studies and crystal structures of copper(II) complexes with H-Aib-X-OH (X=Gly, L-Leu, L-Phe). AB - Synthetic procedures are described that allow access to new copper(II) complexes with dipeptides containing the alpha-aminoisobutyric residue (Aib) as ligands. The solid complexes [Cu(H(-1)L(A))](n).nH(2)O (1) (L(A)H=H-Aib-Gly-OH), [Cu(H( 1)L(B))(MeOH)](n).nMeOH (2) (L(B)H=H-Aib-L-Leu-OH) and [Cu(H(-1)L(C))](n) (3) (L(C)H=H-Aib-L-Phe-OH) have been isolated and characterized by single-crystal X ray crystallography, solid-state IR spectra and UV-Vis spectroscopy in solution (H(-1)L(2-) is the dianionic form of the corresponding dipeptide). Complexes 1 and 3 are three-dimensional coordination polymers with similar structures. The doubly deprotonated dipeptide behaves as a N(amino), N(peptide), O(carboxylate), O'(carboxylate), O(peptide) mu(3) ligand and binds to one Cu(II) atom at its amino and peptide nitrogens and at one carboxylate oxygen, to a second metal at the other carboxylate oxygen, while a third Cu(II) atom is attached to the peptide oxygen. The geometry around copper(II) is distorted square pyramidal with the peptide oxygen at the apex of the pyramid. The structure of 2 consists of zigzag polymeric chains, where the doubly deprotonated dipeptide behaves as a N(amino), N(peptide), O(carboxylate), O'(carboxylate) mu(2) ligand. The geometry at copper(II) is square pyramidal with the methanol oxygen at the apex. The IR data are discussed in terms of the nature of bonding and known structures. The UV Vis spectra show that the solid-state structures of 1, 2 and 3 do not persist in H(2)O. PMID- 12576273 TI - Coordination of thiouridine monophosphates with selected metal ions. AB - Potentiometric and spectroscopic data obtained for the complexes of two thio substituted uridine-monophosphates with Cu(2+), Ni(2+) and Cd(2+) ions have shown that both thionucleotide are more effective ligands than their nucleoside analogues. The basic binding site for all metal ions is the sulfur atom. The chelation by adjacent N(3) donor is also likely, although unfavorable four membered chelate ring is formed. PMID- 12576274 TI - Cytotoxicity and mode of action of vanada- and niobatricarbadecaboranyl monohalide complexes in human HL-60 promyelocytic leukemia cells. AB - Vanada- and niobatricarbadecaboranyl monohalide complexes proved to be potent cytotoxic agents against murine and human leukemia and lymphoma growth as well as HeLa suspended uterine carcinoma. The vanada complex reduced the growth of KB nasopharynx, Hepe liver, HCT-8 ileum and 1-A9 ovary solid carcinomas. A mode of action study in human HL-60 promyelocytic leukemia cells showed that DNA and purine de novo syntheses were significantly inhibited with suppression of the regulatory enzymes activities of DNA polymerase alpha and PRPP-amido transferase. There was moderate inhibition of RNA synthesis and m-RNA polymerase activity. These complexes did not inhibit human topoisomerase I or II activity, although the niobium complex nicked the DNA. The complexes did activate caspases 3, 6 and 9 which are linked to apoptosis programmed cell death. These vanada- and niobatricarbadecaboranyl monohalide complexes appear to be more specific in their effects on leukemia cell metabolism than other sandwich complexes which have broad effects on multiple enzymes. PMID- 12576275 TI - Copper(II) complexes with uridine, uridine 5'-monophosphate, spermidine, or spermine in aqueous solution. AB - Molecular complexes of the types (Urd)H(x)(PA) and (UMP)H(x)(PA) are formed in the uridine (Urd) or uridine 5'-monophosphate (UMP) plus spermidine or spermine systems, as shown by the results of equilibrium and spectral studies. Overall stability constants of the adducts and equilibrium constants of their formation have been determined. An increase in the efficiency of the reaction between the bioligands is observed with increasing length of the polyamine. The pH range of adduct formation is found to coincide with that in which the polyamine is protonated while uridine or its monophosphate is deprotonated. The -NH(x)(+) groups from PA and the N(3) atom of the purine base as well as phosphate groups from the nucleotides have been identified as the significant centres of non covalent interactions. Compared to cytidine, the pH range of Urd adduct formation is shifted significantly higher due to differences in the protonation constants of the endocyclic N(3) donor atoms of particular nucleosides. Overall stability constants of the Cu(II) complexes with uridine and uridine 5'-monophosphate in ternary systems with spermidine or spermine have been determined. It has been found from spectral data that in the Cu(II) ternary complexes with nucleosides and polyamines the reaction of metallation involves mainly N(3) atoms from the pyrimidine bases, as well as the amine groups of PA. This unexpected type of interaction has been evidenced in the coordination mode of the complexes forming in the Cu-UMP systems including spermidine or spermine. Results of spectral and equilibrium studies indicate that the phosphate groups taking part in metallation are at the same time involved in non-covalent interaction with the protonated polyamine. PMID- 12576276 TI - Synthesis, equilibrium studies and structural characterisation of the Zn(II) complexes with trimethylene-N6,N6'-bisadenine. AB - The new compound trimethylene-N(6),N(6')-bisadenine (L), in which two adenine molecules are linked together by a trimethylene bridge that connects the N(6) atoms, has been prepared. Reaction of L with HgCl(2) and ZnCl(2) in concentrated HCl solution leads to crystalline solids. The X-ray characterisation of the Hg(II) complex (H(2)L)[HgCl(4)].3H(2)O reveals that it is an outer-sphere complex in which the ligand is protonated at N(1) and N(1'). In contrast, the structure of the complex [H(2)L(ZnCl(3))(2)].2H(2)O shows the ligand co-ordinated to two different Zn(II) ions through the N(7) of both adenine fragments, the protons being located on the N(1) atoms. The latter compound constitutes the first crystallographic evidence of an inner sphere complex with bis-adenines and, for this reason, an equilibrium study was carried out on the Zn(II)-L-H(+) system. Potentiometric studies indicate that L is protonated in aqueous solution to form HL(+) and H(2)L(2+) with logK(H) values of 4.42 and 3.35 (25 degrees C, 0.10 M KNO(3)). The data from potentiometric titrations in the presence of Zn(2+) can be analysed considering the formation of the species LZn(2+), HLZn(3+), LZn(2)(4+) and HLZn(2)(5+), whose stability constants exceed the value expected for a monodentate interaction of the metal ion with adenine and suggest the possibility of a polydentate behaviour of L in the pH range 2.5-5.0. In contrast, spectrophotometric titrations carried out under conditions similar to those used in the synthetic work (1 M HCl) can be fitted with a model involving exclusively the H(2)LZn(4+) and H(2)LZn(2)(6+) species with logK(M) values reasonable for the interaction of Zn(II) with the N(7) of the protonated adenine fragments. Despite the H(2)LZn(2)(6+) species has a low stability, the spectrophotometric results are in agreement with its formation under the conditions in which the solid complex was prepared. PMID- 12576277 TI - Comparison of substrate metabolism by cytochromes P450 2B1, 2B4, and 2B6: relationship of heme spin state, catalysis, and the effects of cytochrome b5. AB - The metabolism of selected substrates by cytochromes P450 (P450) 2B1, 2B4, and 2B6 was compared, and the effects of cytochrome b(5) (b(5)) on these reactions were assessed. There did not appear to be any trends regarding the effects of b(5) when the metabolism of a given substrate by the different P450 enzymes was compared. The changes in spin states of the P450 enzymes as a result of interactions with substrates and cytochrome b(5) were also determined. Only P450 2B4 demonstrated a relationship between spin state, reaction coupling and b(5) effects. The rates of benzphetamine and 7-ethoxy-4-trifluoromethylcoumarin metabolism by the three enzymes could be correlated with the proportions of high spin heme. Similarly, the proportion of reaction coupling during the metabolism of selected substrates was approximately equal to the proportion of high spin P450. The data are interpreted to indicate that a P450 conformational equilibrium coordinately regulates catalysis and spin state changes. PMID- 12576278 TI - Systematic studies on pH-dependent transformations of dinuclear vanadium(V) citrate complexes in aqueous solutions. A perspective relevance to aqueous vanadium(V)-citrate speciation. AB - Vanadium(V) involvement in interactions with physiological ligands in biological media prompted us to delve into the systematic pH-dependent synthesis, spectroscopic characterization, and perusal of chemical properties of arising aqueous vanadium(V)-citrate species in the requisite system. To this end, facile reactions led to dinuclear complexes (NH(4))(4)[V(2)O(4)(C(6)H(5)O(7))(2)].4H(2)O (1) and (NH(4))(6)[V(2)O(4)(C(6)H(4)O(7))(2)].6H(2)O (2). Complex 1 and 2 were characterized by elemental analysis, FT-IR and X-ray crystallography. Complex 1 crystallizes in the monoclinic space group C2/c with a=16.998(5) A, b=16.768(5) A, c=9.546(3) A, beta=105.22(1) degrees, V=2625(1) A(3), and Z=4. Complex 2 crystallizes in the triclinic space group P1;, with a=9.795(4) A, b=9.942(4) A, c=9.126(3) A, alpha=90.32(1) degrees, beta=111.69(1) degrees, gamma=108.67(1) degrees, V=774.5(5) A(3), and Z=1. The structures of 1 and 2 were consistent with the presence of a V(V)(2)O(2) core, to which citrate ligands of differing protonation state were bound in a coordination mode consistent with past observations. Ultimately, the aqueous pH dependent transformations of a series of three dinuclear complexes, 1, 2 and (NH(4))(2)[V(2)O(4)(C(6)H(6)O(7))(2)].2H(2)O (3), all isolated at pH values from 3 to 7.5, were explored and revealed an important interconnection among all species. Collectively, pH emerged as a determining factor of structural attributes in all three complexes, with the adjoining acid-base chemistry unfolding around the stable V(V)(2)O(2) core. The results point to the participation of all three species in aqueous vanadium(V) citrate speciation, and may relate the site-specific protonations-deprotonations on the dinuclear complexes to potential biological processes involving vanadium(V) and physiological ligand targets. PMID- 12576279 TI - Pentacopper(II) 12-metallacrown-4 complexes with alpha- and beta-aminohydroxamic acids in aqueous solution: a reinvestigation. AB - A reinvestigation of the equilibria of (S)-alpha-alaninehydroxamic acid (alpha Alaha) and (R)-aspartic-beta-hydroxamic acid (Asp-beta-ha) with copper(II) was performed in aqueous solution in order to clarify some contradictory literature reports regarding the stoichiometry of the polynuclear complexes formed. beta Alaninehydroxamic acid (beta-Alaha, HL), for which the formation of a planar 12 metallacrown-4, [Cu(5)L(4)H(-4)](2+), was already reported, was also re-examined for comparison. Among the different techniques used (potentiometry, absorption spectrophotometry, spectropolarimetry and electrospray ionization mass spectrometry), ES data allowed to define unambiguously that all these three ligands form the same pentanuclear species. Therefore it can be concluded that in aqueous solution the hydroxamates of both alpha- and beta-amino acids form 12 metallacrown-4 complexes, and that the formers are less stable. PMID- 12576280 TI - Pt(II) and Pd(II) derivatives of ter-butylsarcosinedithiocarbamate. Synthesis, chemical and biological characterization and in vitro nephrotoxicity. AB - This work reports on the synthesis, characterization and biological activity of new coordination compounds of the type [M(TSDTM)X(2)] (M=Pt(II), Pd(II); X=Cl, Br; TSDTM=ter-butylsarcosine(S-methyl)dithiocarbamate) and [Pd(TSDT)X](n) (TSDT=ter-butylsarcosinedithiocarbamate) in order to study their behavior as potential antitumor agents. All the synthesized compounds were characterized by means of elemental analysis, FT-IR, (1)H and (13)C-NMR spectroscopy and thermogravimetric analysis, suggesting a chelate S,S' structure of the TSDTM/TSDT ligand in a square-planar geometry. Finally, the synthesized complexes have been tested for in vitro cytotoxic activity against human leukemic HL60 and adenocarcinoma HeLa cells; the most active compound [Pt(TSDTM)Br(2)], characterized by IC(50) values very similar to those of the reference compound (cisplatin), was also tested for in vitro nephrotoxicity showing a very low renal cytotoxicity as compared to cisplatin itself. PMID- 12576281 TI - Interaction between macrocyclic nickel complexes and the nucleotides GMP, AMP and ApG. AB - Reactions between the nucleotides GMP, AMP and ApG and the complexes Ni(tren), Ni(cyclam) and NiCR in aqueous solution have been monitored by (1)H, (15)N NMR and UV spectroscopy. The three nickel complexes display different properties in reactions with nucleotides. Ni(tren) which has a pseudo-octahedral coordination geometry was shown to bind to all three nucleotides. Ni(cyclam) and NiCR, both with four nitrogen atoms in a square planar arrangement are not able to bind to nucleotides efficiently because of steric hindrance. Oxidation of Ni(cyclam) by KHSO(5) to produce trivalent Ni(III)(cyclam) improves the coordination capacity, while oxidation of NiCR does not produce a similar effect. The nucleotides interact with trivalent nickel complexes to different extent. Ni(III)CR is seen to oxidize GMP gradually but does not affect AMP significantly. Ni(III)(cyclam), on the other hand, does not oxidize either GMP or AMP at the 1:1 concentration of oxidant used. This result is probably due to the lower redox potential of Ni(cyclam). ApG binds less efficiently to the Ni complexes but is easier oxidized than the mononucleotides. PMID- 12576282 TI - Cooperativity between pepsin and crystallization of calcium carbonate in distilled water. AB - Cooperativity between pepsin and crystallization of calcium carbonate in distilled water was studied. The results show that vaterite was formed under the influence of pepsin and the crystalline product was a composite of vaterite and pepsin. The component of this material was similar to that of nacre. At the same time, the crystallization of calcium carbonate had also an important effect on the secondary structure of the pepsin. The secondary structure of the pepsin was characterized through FT-IR technology. The result indicated that the pure pepsin is composed of 24.38% alpha-helices, 29.91% beta-sheets, 39.32% beta-turns and 6.49% random structures and the pepsin in the CaCO(3)-pepsin solution is composed of 2.09% alpha-helices, 93.304% beta-sheets, 4.592% beta-turns and 0.006% random structure. During the crystallization of the calcium carbonate from the pepsin solution, the secondary structure of the pepsin transformed. These results showed that there was cooperativity between the crystallization of vaterite and the pepsin. The cooperative mechanism is discussed. PMID- 12576283 TI - Interaction of Cu(II)-Arg-Gly-His-Xaa metallopeptides with DNA: effect of C terminal residues, Leu and Glu. AB - The interactions of Cu(II)-Arg-Gly-His-Xaa metallopeptides with DNA (where Xaa is L-Leu or L-Glu) were investigated by DNA-fiber EPR spectroscopy, ESI-MS spectrometry, and agarose gel electrophoresis. The average angle between the g// axis of Cu(II)-Arg-Gly-His-Leu and the DNA-fiber axis increased from 45 degrees at room temperature to 90 degrees at -150 degrees C. The Cu(II)-Arg-Gly-His-Glu complex partly dissociated on DNA to several species. The g//value (2.341) of the main species was smaller than that (2.377) observed for free Cu(II) ion bound to DNA. This indicated that the Cu(II) ion was transferred by the peptide to a DNA site where the free Cu(II) ion can hardly reach. ESI-MS spectra of a mixture of the Cu(II) peptide complex and the oligodeoxynucleotide, [d(CGCGTATACGCG)], suggested that the maximum binding stoichiometries of Cu(II) peptide complexes and double stranded oligodeoxynucleotides were 3:1 for Cu(II)-Arg-Gly-His-Leu and 2:1 for Cu(II)-Arg-Gly-His-Glu, respectively. Cu(II)-Arg-Gly-His-Glu completely converted the supercoiled DNA to the nicked-circular form, whereas the cleavage activity was considerably reduced when excess ligand was added. In the presence of excess peptide, nicked DNA formation ratios were 64% for Cu(II)-Arg-Gly-His Leu and 15% for Cu(II)-Arg-Gly-His-Glu, respectively. The negative charge on Cu(II)-Arg-Gly-His-Glu reduced the affinity of the complex for DNA and enhanced the specificity of the binding. PMID- 12576284 TI - Novel oxorhenium and oxotechnetium MO(NS)(S)2 complexes in the development of 5 HT1A receptor imaging agents. AB - The [NS][S](2) mixed-ligand system was applied to synthesize oxorhenium and oxotechnetium complexes of the general formula MO(o CH(3)OC(6)H(4)N(CH(2)CH(2))(2)NCH(2)CH(2)S)(p-CH(3)C(6)H(4)S)(2) (M=Re in 1, M=(99)Tc in 2, and M=(99m)Tc in 3). The bidentate [NS] ligand includes the 1-(2 methoxyphenyl)piperazine moiety which is a fragment of the true 5-HT(1A) antagonist WAY 100635. The oxorhenium complex 1 was prepared by a ligand exchange reaction using ReOCl(3)(PPh(3))(2) as precursor while [Bu(4)N][(99)TcOCl(4)] and (99)Tc-gluconate were used as precursors in the synthesis of the oxotechnetium-99 complex 2. Both complexes were characterized by elemental analysis and spectroscopic methods. Crystallographic analysis of 1 showed that the rhenium coordination geometry is trigonal bipyramidal. The basal plane of the trigonal bipyramid is defined by the oxo group and two sulphur atoms, one belonging to the [NS] ligand and the other to an aromatic thiol, while the apical positions are occupied by the nitrogen of the [NS] ligand and the sulphur of the second aromatic thiol. The oxotechnetium-99 complex 2 has almost identical unit cell parameters to those of the oxorhenium complex 1 indicating, in combination with the other analytical data, that the complexes are isostructural. The binding affinity of the oxorhenium complex 1 for the 5-HT(1A) receptor subtype was determined in rat brain hippocampal preparations (IC(50)=106 nM). The oxotechnetium-99m complex 3 was prepared by a ligand exchange reaction using (99m)Tc-glucoheptonate as the precursor. Its structure was established by comparative HPLC studies using the oxotechnetium-99 complex 2 as a reference. Complex 3 was administered by intravenous injection in rats. At 2 min post injection, 0.153% of the injected dose per gram of tissue was measured in rat brain. PMID- 12576285 TI - Enantioselective binding of lambda- and delta-[Ru(bpy)(2)(HPIP)]Cl2 (HPIP=2-(2 hydroxyphenyl)imidazo[4,5-f][1,10]phenanthroline) to the hexanucleotide [d(5' GTCGAC-3')2]. AB - Multidimensional NMR techniques (1D (1)HNMR, 2D DQF (1)H(1)H COSY and 2D (1)H(1)H NOESY), electrospray ionization mass spectrometry (ESI-MS) and electronic spectroscopy, were performed to study the interactions of the enantiomers lambda- and delta-[Ru(bpy)(2)(HPIP)]Cl(2), (HPIP=2-(2-hydroxyphenyl)imidazo[4,5 f][1,10]phenanthro-line) with the self complementary hexanucleotide duplex d(5' GTCGAC-3')(2). The results show that the delta-[Ru(bpy)(2)(HPIP)]Cl(2) binds tightly to the oligonucleotide, by intercalation of the ligand HPIP, between the A5 and C6 base sequence of the same strand, probably through the minor groove. Lambda-enantiomer binds weakly, suggesting groove interactions with the hexanucleotide duplex. ESI-MS spectrometry and UV-vis spectroscopy also confirmed these observations. PMID- 12576286 TI - The catalytic effect of Mg2+ and imidazole on the decomposition of 5 phosphoribosyl-alpha-1-pyrophosphate in aqueous solution. AB - The rate of decomposition of aqueous solutions of 5-phosphoribosyl-alpha-1 pyrophosphate (PRPP) is greatly enhanced by the presence of certain divalent ions. High levels of Mg(2+) were shown to increase the rate of phosphate removal from PRPP by factors greater than 100-fold. The combined action of Mg(2+) and a nitrogenous compound (imidazole was employed in this study) increases the degradation rate constants by up to 1000-fold. The degradation of PRPP in neutral solutions follows two main paths, a pyrophosphate hydrolysis reaction yielding ribose 5-phosphate and an internal beta-phosphate cleavage yielding 5 phosphoribosyl-1,2 cyclic phosphate. The catalytic effect of Mg(2+) on PRPP degradation appears to suggest a connection between the observed rate enhancements and the dimagnesium form of PRPP. Rate enhancing effects of Mg(2+) are greatest at pH values (i.e. pH>7) which promote strong phosphate-Mg(2+) interaction. PMID- 12576287 TI - Substrate inhibition of rat liver and kidney arginase with fluoride. AB - Fluoride is an uncompetitive inhibitor of rat liver arginase. This study has shown that fluoride caused substrate inhibition of rat liver arginase at substrate concentrations above 4 mM. Rat kidney arginase was more sensitive to inhibition by fluoride than liver arginase. For both liver and kidney arginase preincubation with fluoride had no effect on the inhibition. When assayed with various concentrations of L-arginine, rat kidney arginase did not have Michaelis Menten kinetics. Lineweaver-Burk and Eadie-Hofstee plots were nonlinear. Kidney arginase showed strong substrate activation at concentrations of L-arginine above 4 mM. Within narrow concentrations of L-arginine, the inhibition of kidney arginase by fluoride was uncompetitive. Fluoride caused substrate inhibition of kidney arginase at L-arginine concentrations above 1 mM. The presence of fluoride prevented the substrate activation of rat kidney arginase. PMID- 12576288 TI - Syntheses, characterization and DNA-binding studies of ruthenium(II) terpyridine complexes: [Ru(tpy)(PHBI)]2+ and [Ru(tpy)(PHNI)]2+. AB - Two novel tridentate ligands, 2-(2-benzimidazole)-1,10-phenanthroline (PHBI) and 2-(2-naphthoimidazole)-1,10-phenanthroline (PHNI), and their heteroleptic complexes [Ru(tpy)(PHBI)](ClO(4))(2).2H(2)O (1) and [Ru(tpy)(PHNI)](ClO(4))(2).H(2)O (2) (tpy=2,2':6',2"-terpyridyl) have been synthesized and characterized by elemental analysis, mass spectra, 1H NMR, and electronic spectroscopy. The electrochemical behaviors of the two novel complexes were studied by cyclic voltammetry. The DNA-binding properties of the two complexes were investigated by spectroscopic methods and viscosity measurements. The results indicated that the two complexes interact with DNA in different binding modes. Complex 1 may bind to DNA via electrostatic interaction, while complex 2 binds to DNA by partial intercalation via the extended naphthyl ring into the base pairs of DNA. PMID- 12576289 TI - High nuclearity nickel compounds with three, four or five metal atoms showing antibacterial activity. AB - The effect on DNA and the antibacterial activity of a series of high nuclearity nickel compounds with three, four and five metal atoms were examined. The compounds have a mixed ligand composition with salicylhydroxamic acid and di-2 pyridyl-ketonoxime as chelate agents. In the trinuclear compound Ni(3)(shi)(2)(Hpko)(2)(py)(2)(1), two metal ions show a square planar geometry while the third one is in an octahedral environment. The compounds with four and five nickel atoms construct metallacrown cores with two distinct connectivities. The tetranuclear vacant metallacrown [12-MC(Ni(II)N(Hshi)2(pko)2)-4](2+) shows the connectivity pattern [-O-Ni-O-N-Ni-N-](2), while the pentanuclear ([Ni(II)][12-MC(Ni(II)N(shi)2(pko)2)-4])(2+) follows the pattern [-Ni-O-N-](4). Two distinct arrangements of the chelates around the ring metal ions were observed; a 6-5-6-5-6-5-6-5 arrangement for the [12-MC(Ni(II)N(Hshi)2(pko)2)-4] core and a 6-6-5-5-6-6-5-5 arrangement for the [12-MC(Ni(II)N(shi)2(pko)2)-4] core. Magnetic variable temperature susceptibility study of the trinuclear compound revealed the presence of one paramagnetic nickel(II) ion with strong crystal field dependence, with D=5.0(4) cm(-1), g(xy)=2.7(3) and g(z)=2.3(3). The effect of the synthesized Ni(II) complexes on the integrity and electrophoretic mobility of nucleic acids was examined. Only compounds 2, 3 and 4 altered the mobility of pDNA, forming high molecular weight concatamers at low concentrations or precipitates at higher concentrations. Antibacterial activity screening of the above compounds suggests that nickel compounds 2, 3 and 4 were the most active and can act as potent antibacterial agents. PMID- 12576290 TI - Synthesis, characterization, and molecular structure of a gallium(III) complex of an amine-phenol ligand with activity against chloroquine-sensitive Plasmodium falciparum strains. AB - Emergence of chloroquine-resistant Plasmodium falciparum strains necessitates discovery of novel antimalarial drugs, especially if the agents can be synthesized from commercially available, inexpensive precursors via short synthetic routes. While exploring structure-activity relationships, we found a gallium(III) complex, [(1,12-bis(2-hydroxy-5-methoxybenzyl)-1,5,8,12 tetraazadodecane)-gallium(III)](+) [Ga-5-Madd](+), 1, that possessed antimalarial efficacy. Like previously reported complexes, the crystal structure of 1 revealed gallium(III) in a symmetrical octahedral environment surrounded by four secondary amine nitrogen atoms in equatorial plane and two axial oxygen atoms. In contrast to a previously reported complex, [Ga-3-Madd](+), this novel metallo-antimalarial 1 possessed modest efficacy against chloroquine-sensitive HB3 Plasmodium lines. Thus, slight variation in the positions of methoxy functionalities on the aromatic rings of the organic scaffold dramatically altered specificity thereby suggesting a targeted (e.g., transporter- or receptor-mediated) rather than non specific (e.g., pH or other gradient-mediated) mechanism of action for these agents. PMID- 12576291 TI - Oxidative cleavage of DNA by tridentate copper (II) complex. AB - Copper (II) complex 1 having planar tridentate ligand, bzimpy, where bzimpy is 2,6-bis(benzimidazo-2-yl) pyridine was synthesized and characterized by UV visible, FAB (fast atom bombardment) mass and infrared spectroscopy. From absorption titration data, the binding constant of Cu(II) with DNA was calculated to be (1.8+/-0.02)x10(4) M(-1). Thermal denaturation study of DNA with 1 revealed deltaT(m) of 5+/-0.5 degrees C. Viscosity measurement showed that complex binds with DNA through intercalative mode. Copper (II) complex induces cleavage in plasmid DNA in the presence of coreductants such as ascorbic acid or glutathione. PMID- 12576292 TI - Adenosine receptors in the nervous system: pathophysiological implications. AB - Adenosine is a ubiquitous homeostatic substance released from most cells, including neurones and glia. Once in the extracellular space, adenosine modifies cell functioning by operating G-protein-coupled receptors (GPCR; A(1), A(2A), A(2B), A(3)) that can inhibit (A(1)) or enhance (A(2)) neuronal communication. Interactions between adenosine receptors and other G-protein-coupled receptors, ionotropic receptors and receptors for neurotrophins also occur, and this might contribute to a fine-tuning of neuronal function. Manipulations of adenosine receptors influence sleep and arousal, cognition and memory, neuronal damage and degeneration, as well as neuronal maturation. These actions might have therapeutic implications for neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, as well as for other neurological situations such as epilepsy, idiopathic pain or even drug addition. Peripheral side effects associated with adenosine receptor agonists limit their usefulness in therapeutics; in contrast, adenosine receptor antagonists appear to have less side effects as it is the case of the well-known non-selective antagonists theophylline (present in tea) or caffeine (abundant in coffee and tea), and their emerging beneficial actions in Parkinson's disease and Alzheimer's disease are encouraging. A(1) receptor antagonism may also be useful to enhance cognition and facilitate arousal, as well as in the periphery when deficits of neurotransmitter release occur (e.g. myasthenic syndromes). Enhancement of extracellular adenosine levels through drugs that influence its metabolism might prove useful approaches in situations such as neuropathic pain, where enhanced activation of inhibitory adenosine A(1) receptors is beneficial. One might then consider adenosine as a fine-tuning modulator of neuronal activity, which via subtle effects causes harmonic actions on neuronal activity. Whenever this homeostasis is disrupted, pathology may be installed and selective receptor antagonism or agonism required. PMID- 12576293 TI - Wiring of the brain by a range of guidance cues. AB - During development of the central nervous system, growth cones navigate along specific pathways, recognize their targets and then form synaptic connections by elaborating terminal arbors. To date, a number of developmental and in vitro studies have characterized the nature of the guidance cues that underlie various types of axonal behavior, from initial outgrowth to synapse formation, including pathway selection, polarized growth, orientated growth, termination and branching. New approaches in molecular biology have identified several types of guidance cues, most of which are likely to act as local cues. Moreover, recent studies have indicated that axonal responsiveness to guidance cues changes dynamically, which appears to be elicited by environmental factors encountered by the navigating growth cones. This article addresses what molecular cues are responsible for guidance mechanisms including axonal responsiveness, focusing on axonal behavior in the developmental stages. PMID- 12576294 TI - Fundamental mechanisms of visual motion detection: models, cells and functions. AB - Taking a comparative approach, data from a range of visual species are discussed in the context of ideas about mechanisms of motion detection. The cellular basis of motion detection in the vertebrate retina, sub-cortical structures and visual cortex is reviewed alongside that of the insect optic lobes. Special care is taken to relate concepts from theoretical models to the neural circuitry in biological systems. Motion detection involves spatiotemporal pre-filters, temporal delay filters and non-linear interactions. A number of different types of non-linear mechanism such as facilitation, inhibition and division have been proposed to underlie direction selectivity. The resulting direction-selective mechanisms can be combined to produce speed-tuned motion detectors. Motion detection is a dynamic process with adaptation as a fundamental property. The behavior of adaptive mechanisms in motion detection is discussed, focusing on the informational basis of motion adaptation, its phenomenology in human vision, and its cellular basis. The question of whether motion adaptation serves a function or is simply the result of neural fatigue is critically addressed. PMID- 12576295 TI - Urokinase potentiates PDGF-induced chemotaxis of human airway smooth muscle cells. AB - We investigated the chemotactic action of PDGF and urokinase on human airway smooth muscle (HASM) cells in culture. Cells were put in collagen-coated transwells with 8-micro m perforations, incubated for 4 h with test compounds, then fixed, stained, and counted as migrated nuclei by microscopy. Cells from all culture conditions showed some basal migration (migration in the absence of stimuli during the assay), but cells preincubated for 24 h in 10% FBS or 20 ng/ml PDGF showed higher basal migration than cells quiesced in 1% FBS. PDGF(BB), PDGF(AA), and PDGF(AB) were all chemotactic when added during the assay. PDGF chemotaxis was blocked by the phosphatidyl 3'-kinase inhibitor LY-294002, the MEK inhibitor U-0126, PGE(2), formoterol, pertussis toxin, and the Rho kinase inhibitor Y-27632. Urokinase alone had no stimulatory effect on migration of quiescent cells but caused a dose-dependent potentiation of chemotaxis toward PDGF. Urokinase also potentiated the elevated basal migration of cells pretreated in 10% FBS or PDGF. This potentiating effect of urokinase appears to be novel. We conclude that PDGF and similar cytokines may be important factors in airway remodeling by redistribution of smooth muscle cells during inflammation and that urokinase may be important in potentiating the response. PMID- 12576296 TI - Ceramide accumulation precedes caspase-3 activation during apoptosis of A549 human lung adenocarcinoma cells. AB - Ceramide, the basic structural unit of sphingolipids, controls the balance between cell growth and death by inducing apoptosis. We have previously shown that accumulation of ceramide, triggered by hydrogen peroxide (H(2)O(2)) or by short-chain ceramide analogs, induces apoptosis of lung epithelial cells. Here we elucidate the link between caspase-3 activation, at the execution phase, and ceramide accumulation, at the commitment phase of apoptosis in A549 human lung adenocarcinoma cells. The induction of ceramide accumulation by various triggers of ceramide generation, such as H(2)O(2), C(6)-ceramide, or UDP-glucose-ceramide glucosyltransferase inhibitor dl-threo-1-phenyl-2-decanoylamino-3-morpholino-1 propanol, triggered the activation of caspase-3. This ceramide elevation also induced the cleavage of the death substrate poly(ADP-ribose) polymerase and was followed by apoptotic cell death. Ceramide-mediated apoptosis was blocked by a general caspase inhibitor, Boc-d-fluoromethylketone, and by overexpression of the antiapoptotic protein Bcl-2. Notably, overexpression of Bcl-2 reduced the basal cellular levels of ceramide and prevented the induction of ceramide generation by C(6)-ceramide, which implies ceramide generation as a possible target for the antiapoptotic effects of Bcl-2. PMID- 12576297 TI - Transcription factor USF2 is developmentally regulated in fetal lung and acts together with USF1 to induce SP-A gene expression. AB - Expression of the pulmonary surfactant protein A (SP-A) gene is lung specific, developmentally regulated, and enhanced by hormones and factors that increase cAMP. We previously identified two E-box-like enhancers termed distal binding element (DBE) and proximal binding element (PBE) in the 5'-flanking region of the rabbit (r) SP-A gene that are essential for cAMP induction of rSP-A promoter activity (Gao E, Alcorn JL, and Mendelson CR. J Biol Chem 268: 19697-19709, 1993). We also found that DBE and PBE serve as binding sites for the basic helix loop-helix-leucine zipper transcription factor, upstream stimulatory factor-1 (USF1) (Gao E, Wang Y, Alcorn JL, and Mendelson CR. J Biol Chem 272: 23398-23406, 1997). In the present study, PBE was used to screen a rabbit fetal lung cDNA expression library; a cDNA insert encoding the structurally related rabbit upstream stimulatory factor-2 (rUSF2) was isolated. The levels of rUSF2 mRNA reach peak levels in fetal rabbit lung at 28 days of gestation, in concert with the time of maximal induction of SP-A gene transcription. In yeast two-hybrid analysis, rUSF2 was found to preferentially form heterodimers, compared with homodimers, with rUSF1. Binding complexes of nuclear proteins isolated from fetal rabbit lung type II cells with the DBE and PBE were supershifted by anti-rUSF2 antibodies. Binding activity was enriched in nuclear proteins from type II cells compared with fibroblasts. Overexpression of rUSF2 in transfected lung A549 cells increased rSP-A promoter activity and acted synergistically with rUSF1. We suggest that heterodimers of USF2 and USF1 bound to two E-box elements in the SP A gene 5'-flanking region serve a key role in developmental and hormonal regulation of SP-A gene expression in pulmonary type II cells. PMID- 12576298 TI - DMT1 expression is increased in the lungs of hypotransferrinemic mice. AB - Despite a lack of transferrin, hypotransferrinemic (Hp) mice demonstrate an accumulation of iron in peripheral organs including the lungs. One potential candidate for such transferrin-independent uptake of iron is divalent metal transporter-1 (DMT1), an established iron transporter. We tested the hypothesis that increased concentrations of iron in the lungs of Hp mice are associated with elevations in DMT1 expression. With the use of inductively coupled plasma emission spectroscopy, measurements of nonheme iron confirmed significantly elevated concentrations in the lung tissue of Hp mice relative to the wild-type mice. Western blot analyses for the expression of two isoforms of DMT1 in the Hp mice relative to the wild-type animals demonstrated an elevation for the isoform that lacks an iron-responsive element (IRE) with significant decrements in the expression of +IRE DMT1. With the use of immunohistochemistry, -IRE DMT1 was localized to both airway epithelial cells and alveolar macrophages in wild-type mice. Staining appeared increased in both types of cells in the Hp mice. Elevated concentrations of both tissue nonheme iron and expression of -IRE DMT1 in the Hp mice were associated with increased quantities of -IRE mRNA. There was no difference between wild-type and homozygotic Hp mice in the amount of mRNA for DMT1 +IRE. We conclude that differences between Hp and wild-type mice in nonheme iron concentrations were accompanied by increases in the expression of -IRE DMT1. Increased expression of -IRE DMT1 in the lungs of the Hp mice could be responsible for elevated concentrations of the metal in these tissues. PMID- 12576299 TI - Transient injury to rat lung mitochondrial DNA after exposure to hyperoxia and inhaled nitric oxide. AB - The effect of hyperoxia alone and in combination with inhaled nitric oxide (NO) on the integrity of lung mitochondrial DNA (mtDNA) in vivo was evaluated in Fischer 344 rats. PCR amplification of lung mtDNA using two sets of primers spanning 10.1 kb of the mtDNA revealed that inhalation of 20 ppm of NO in conjunction with hyperoxia (>95% O2) reduced the amplification of mtDNA templates by 10 +/- 1% and 26 +/- 3% after 24 h of exposure. The ability of mtDNA to amplify was not compromised in rats exposed to 80% O2, even in the presence of 20 ppm of inhaled NO. Surprisingly, exposure to >95% O2 alone for either 24 or 48 h did not compromise the integrity of mtDNA templates compared with air-exposed controls, despite evidence of genomic DNA injury. Interestingly, inhaling NO alone for 48 h increased mtDNA amplification by 12 +/- 2% to 21 +/- 7%. Injury to the lung mtDNA after exposure to >95% O2 plus 20 ppm of NO was transient as rats allowed to recover in room air after exposure displayed increased amplification, with levels exceeding controls by 20 +/- 3% to 29 +/- 4%. Increased amplification was not due to cellular proliferation or increased mitochondrial number. Moreover, the ratio of pulmonary mtDNA to genomic DNA remained the same between treatment groups. The results indicate that hyperoxia fails to induce significant injury to mtDNA, and whereas inhalation of NO with hyperoxia results in mtDNA damage, the lesions are rapidly repaired during recovery. PMID- 12576301 TI - Bile acid regulation of hepatic physiology: III. Bile acids and nuclear receptors. AB - Bile acids are physiological detergents that facilitate excretion, absorption, and transport of fats and sterols in the intestine and liver. Recent studies reveal that bile acids also are signaling molecules that activate several nuclear receptors and regulate many physiological pathways and processes to maintain bile acid and cholesterol homeostasis. Mutations of the principal regulatory genes in bile acid biosynthetic pathways have recently been identified in human patients with hepatobiliary and cardiovascular diseases. Genetic manipulation of key regulatory genes and bile acid receptor genes in mice have been obtained. These advances have greatly improved our understanding of the molecular mechanisms underlying complex liver physiology but also raise many questions and controversies to be resolved. These developments will lead to early diagnosis and discovery of drugs for treatment of liver and cardiovascular diseases. PMID- 12576300 TI - Diesel exhaust particles upregulate eotaxin gene expression in human bronchial epithelial cells via nuclear factor-kappa B-dependent pathway. AB - Fine particles derived from diesel engines, diesel exhaust particles (DEP), have been shown to augment gene expression of several inflammatory cytokines in human airway epithelial cells in vitro. However, it remains unclear whether or not DEP have any effect on the expression and production of eotaxin, an important chemokine involved in eosinophil recruitment into the airways. We studied the effects of DEP by using a conventional suspended DEP and by a recently established in vitro cell exposure system to diesel exhaust (Abe S, Takizawa H, Sugawara I, and Kudoh S, Am J Respir Cell Mol Biol 22: 296-303, 2000). DEP showed a dose-dependent stimulatory effect on eotaxin production by normal human peripheral airway epithelial cells as well as by bronchial epithelial cell line BET-1A as assessed by specific ELISA. mRNA levels increased by DEP were shown by RT-PCR. DEP showed an additive effect on IL-13-stimulated eotaxin expression. DEP induced NF-kappaB activation by EMSA as previously reported but did not induce signal transducer and activator of transcription (STAT) 6 activation according to Western blot analysis. Finally, antioxidant agents (N-acetyl cysteine and pyrrolidine dithiocarbamate), which inhibited NF-kappaB activation but failed to affect STAT6 activation, almost completely attenuated DEP-induced eotaxin production, whereas these agents failed to attenuate IL-13-induced eotaxin production. These findings suggested that DEP stimulated eotaxin gene expression via NF-kappaB-dependent, but STAT6-independent, pathways. PMID- 12576302 TI - Musings on the wanderer: what's new in our understanding of vago-vagal reflex? IV. Current concepts of vagal efferent projections to the gut. AB - Vagal efferents, consisting of distinct lower motor and preganglionic parasympathetic fibers, constitute the motor limb of vagally mediated reflexes. Arising from the nucleus ambiguus, vagal lower motor neurons (LMN) mediate reflexes involving striated muscles of the orad gut. LMNs provide cholinergic innervation to motor end plates that are inhibited by myenteric nitrergic neurons. Preganglionic neurons from the dorsal motor nucleus implement parasympathetic motor and secretory functions. Cholinergic preganglionic neurons form parallel inhibitory and excitatory vagal pathways to smooth muscle viscera and stimulate postganglionic neurons via nicotinic and muscarinic receptors. In turn, the postganglionic inhibitory neurons release ATP, VIP, and NO, whereas the excitatory neurons release ACh and substance P. Vagal motor effects are dependent on the viscera's intrinsic motor activity and the interaction between the inhibitory and excitatory vagal influences. These interactions help to explain the physiology of esophageal peristalsis, gastric motility, lower esophageal sphincter, and pyloric sphincter. Vagal secretory pathways are predominantly excitatory and involve ACh and VIP as the postganglionic excitatory neurotransmitters. Vagal effects on secretory functions are exerted either directly or via release of local mediators or circulating hormones. PMID- 12576303 TI - Development of the myogenic response in postnatal intestine: role of PKC. AB - Previous attempts to determine developmental changes in the vascular myogenic response have been confounded by the presence of competing vasoactive stimuli or the use of isolated vessels with markedly different baseline diameters. To circumvent these issues, small mesenteric arteries (diameter approximately 150 microm) from 1- and 10-day-old piglets were studied in vitro under no-flow conditions. In situ studies demonstrated that the intravascular pressure and diameter of these vessels were similar in both age groups, allowing an effective comparison of the myogenic response not obscured by differences in basal diameter. The pressure-diameter relationship was age specific. Thus, although small mesenteric arteries from both age groups demonstrated myogenic constriction in response to stepwise increases in pressure (0 to 100 mmHg, in 20-mmHg increments), the intensity of contraction was significantly greater in vessels from 1-day-old piglets particularly within the pressure range normally experienced by these vessels in situ. Attenuation or activation of PKC with calphostin C or indolactam, respectively, substantially altered the pressure diameter relationship in 1-, but not 10-day-old arteries; thus calphostin C essentially eliminated the contractile response to pressure elevation in younger subjects, whereas indolactam significantly increased the intensity of the myogenic response and shifted its activation point to a lower pressure range. Immunoblots carried out on protein recovered from these arteries revealed the presence of alpha, beta, epsilon, iota, and lambda; notably, expression of the alpha- and epsilon-isoforms substantially decreased between postnatal days 1 and 10. PMID- 12576304 TI - Erk1/2- and p38 MAP kinase-dependent phosphorylation and activation of cPLA2 by m3 and m2 receptors. AB - This study examined the upstream signaling pathways initiated by muscarinic m2 and m3 receptors that mediate sustained ERK1/2- and p38 MAP kinase-dependent phosphorylation and activation of the 85-kDa cytosolic phospholipase (cPL)A(2) in smooth muscle. The pathway initiated by m2 receptors involved sequential activation of Gbetagamma(i3), phosphatidylinositol (PI)3-kinase, Cdc42, and Rac1, p21-activated kinase (PAK1), p38 mitogen-activated protein (MAP) kinase, and cPLA(2), and phosphorylation of cPLA(2) at Ser(505). cPLA(2) activity was inhibited to the same extent (61 +/- 5 to 72 +/- 4%) by the m2 antagonist methoctramine, Gbeta antibody, pertussis toxin, the PI3-kinase inhibitor LY 294002, PAK1 antibody, the p38 MAP kinase inhibitor SB-203580, and a Cdc42/Rac1 GEF (Vav2) antibody and by coexpression of dominant-negative Cdc42 and Rac1 mutants. The pathway initiated by m3 receptors involved sequential activation of Galpha(q), PLC-beta1, PKC, ERK1/2, and cPLA(2), and phosphorylation of cPLA(2) at Ser(505). cPLA(2) activity was inhibited to the same extent (35 +/- 3 to 41 +/- 5%) by the m3 antagonist 4-diphenylacetoxy-N-methylpiperdine (4-DAMP), the phosphoinositide hydrolysis inhibitor U-73122, the PKC inhibitor bisindolylmaleimide, and the ERK1/2 inhibitor PD 98059. cPLA(2) activity was not affected in cells coexpressing dominant-negative RhoA and PLC-delta1 mutants, implying that PKC was not derived from phosphatidylcholine hydrolysis. The effects of ERK1/2 and p38 MAP kinase on cPLA(2) activity were additive and accounted fully for activation and phosphorylation of cPLA(2). PMID- 12576305 TI - Effects of ginger on motion sickness and gastric slow-wave dysrhythmias induced by circular vection. AB - Ginger has long been used as an alternative medication to prevent motion sickness. The mechanism of its action, however, is unknown. We hypothesize that ginger ameliorates the nausea associated with motion sickness by preventing the development of gastric dysrhythmias and the elevation of plasma vasopressin. Thirteen volunteers with a history of motion sickness underwent circular vection, during which nausea (scored 0-3, i.e., none to severe), electrogastrographic recordings, and plasma vasopressin levels were assessed with or without ginger pretreatment in a crossover-design, double-blind, randomized placebo-controlled study. Circular vection induced a maximal nausea score of 2.5 +/- 0.2 and increased tachygastric activity and plasma vasopressin. Pretreatment with ginger (1,000 and 2,000 mg) reduced the nausea, tachygastria, and plasma vasopressin. Ginger also prolonged the latency before nausea onset and shortened the recovery time after vection cessation. Intravenous vasopressin infusion at 0.1 and 0.2 U/min induced nausea and increased bradygastric activity; ginger pretreatment (2,000 mg) affected neither. Ginger effectively reduces nausea, tachygastric activity, and vasopressin release induced by circular vection. In this manner, ginger may act as a novel agent in the prevention and treatment of motion sickness. PMID- 12576306 TI - Role of nerve growth factor in modulation of gastric afferent neurons in the rat. AB - Recent studies demonstrated that experimental ulcers are associated with changes in the properties of voltage-sensitive sodium currents in sensory neurons. We hypothesized that nerve growth factor (NGF) contributes to these changes. Gastric ulcers were induced by acetic acid injection into the wall of the rat stomach. NGF expression was determined by ELISA and immunohistochemically. Sensory neurons were labeled by injection of a retrograde tracer into the gastric wall. Sodium currents were recorded in gastric sensory neurons from nodose and dorsal root ganglia cultured for 24 h in the presence of NGF or a neutralizing NGF antibody, respectively. Gastric ulcer formation caused a rise in NGF concentration within the gastric wall and an increase in NGF immunoreactivity. Exposure to NGF caused a significant increase in the TTX-resistant sodium current, whereas the TTX sensitive sodium current remained unchanged. This was associated with an acceleration of the recovery from inactivation in spinal sensory neurons. Production and release of NGF in the gastric wall may contribute to sensitization of primary afferent neurons during gastric inflammation. PMID- 12576307 TI - Complement receptor 1 inhibitors for prevention of immune-mediated red cell destruction: potential use in transfusion therapy. AB - Activation of complement cascade via the antibody-mediated classical pathway can initiate red blood cell (RBC) destruction, causing transfusion reactions and hemolytic anemia. In the present study, we have assessed the ability of a human recombinant soluble form of complement receptor 1 (sCR1) to inhibit complement mediated RBC destruction in vitro and in vivo. Using an in vitro alloimmune incompatibility model, sCR1 inhibited complement activation and prevented hemolysis. Following transfusion of human group O RBCs into mice lacking detectable pre-existing antibodies against the transfused RBCs, systemic coadministration of 10 mg/kg sCR1, a dose well tolerated in human subjects for prevention of tissue injury, completely inhibited the in vivo clearance of the transfused RBCs and surface C3 deposition in the first hour after transfusion, correlating with the half-life of sCR1 in the circulation. Treatment with sCR1 increased the survival of transfused human group A RBCs in the circulation of mice with pre-existing anti-A for 2 hours after transfusion by 50%, reduced intravascular hemolysis, and lowered the levels of complement deposition (C3 and C4), but not immunoglobulin G (IgG) or IgM, on the transfused cells by 100-fold. We further identified potential functional domains in CR1 that can act to limit complement-mediated RBC destruction in vitro and in vivo. Collectively, our data highlight a potential use of CR1-based inhibitors for prevention of complement dependent immune hemolysis. PMID- 12576308 TI - Glycophorin C is the receptor for the Plasmodium falciparum erythrocyte binding ligand PfEBP-2 (baebl). AB - We report in this paper that glycophorin C (GPC) is the receptor for PfEBP-2 (baebl, EBA-140), the newly identified erythrocyte binding ligand of Plasmodium falciparum. PfEBP-2 is a member of the Duffy binding-like erythrocyte binding protein (DBL-EBP) family. Although several reports have been published characterizing PfEBP-2, the identity of its erythrocytic receptor was still unknown. Using a combination of enzymatically treated red blood cells (RBCs) and rare, variant RBCs lacking different surface proteins, we have shown that PfEBP-2 does not bind to cells lacking GPC. Additionally, we found that PfEBP-2 binds differentially to variants of GPC lacking exon 2 or exon 3, and determined that the binding domain on GPC is potentially restricted to amino acid residues 14 through 22 within exon 2. Thus PfEBP-2 is involved in a sialic acid-dependent pathway of invasion, which does not involve glycophorin A or glycophorin B and represents a novel route of entry into the RBCs. PMID- 12576309 TI - Tumor-derived, chaperone-rich cell lysate activates dendritic cells and elicits potent antitumor immunity. AB - We have utilized a free-solution isoelectric focusing technique (FS-IEF) to obtain chaperone-rich cell lysate (CRCL) fractions from clarified tumor homogenates and have previously reported on their vaccinating potential. To better understand the underlying mechanisms as well as to improve on the immunizing efficacy of tumor-derived chaperone complexes, in the present study we examined the effects of CRCL-loaded dendritic cells (DCs) against 12B1, an aggressive bcr-abl+ murine leukemia tumor. We found that DCs incubated with 12B1 derived CRCL had higher expression of CD40 and major histocompatibility complex class II (MHC-II) on their cell surface, produced more interleukin-12 (IL-12), and had superior immunostimulatory capacity in a mixed leukocyte reaction (MLR) when compared with DCs exposed to unfractionated tumor lysate or purified heat shock protein 70 (HSP70). Vaccination of mice with 12B1 CRCL-pulsed DCs significantly prolonged their survival, with more than 80% of mice rejecting their tumors following a lethal challenge with live 12B1 compared with those immunized with tumor lysate or HSP70-loaded DCs. The protective immunity generated was tumor specific, long lasting, and both CD4+ and CD8+ T-cell dependent. Moreover, immunization with CRCL-loaded DCs resulted in a 75% cure rate in mice with pre-existing 12B1 tumors. Our findings indicate that CRCL has prominent adjuvant effects and is a very effective source of tumor antigen for pulsing DCs. FS-IEF-derived CRCL-pulsed DCs are a promising anticancer vaccine that warrants clinical research and development. PMID- 12576310 TI - Apoptotic neutrophils in the circulation of patients with glycogen storage disease type 1b (GSD1b). AB - Glycogen storage disease type 1b (GSD1b) is a rare autosomal recessive disorder characterized by hypoglycemia, hepatomegaly, and growth retardation, and associated-for unknown reasons- with neutropenia and neutrophil dysfunction. In 5 GSD1b patients in whom nicotin-amide adenine dinucleotide phosphate-oxidase activity and chemotaxis were defective, we found that the majority of circulating granulocytes bound Annexin-V. The neutrophils showed signs of apoptosis with increased caspase activity, condensed nuclei, and perinuclear clustering of mitochondria to which the proapoptotic Bcl-2 member Bax had translocated already. Granulocyte colony-stimulating factor (G-CSF) addition to in vitro cultures did not rescue the GSD1b neutrophils from apoptosis as occurs with G-CSF-treated control neutrophils. Moreover, the 2 GSD1b patients on G-CSF treatment did not show significantly lower levels of apoptotic neutrophils in the bloodstream. Current understanding of neutrophil apoptosis and the accompanying functional demise suggests that GSD1b granulocytes are dysfunctional because they are apoptotic. PMID- 12576311 TI - Dysfunction and infection of freshly isolated blood myeloid and plasmacytoid dendritic cells in patients infected with HIV-1. AB - Recently it has been shown that the 2 populations of blood dendritic cells (DCs), termed plasmacytoid (pcDCs) and myeloid (myDCs), are reduced in HIV-1 infection. This study aimed to determine whether these 2 populations are targets for HIV-1 infection and whether their ability to stimulate T-lymphocyte proliferation is affected. Highly purified populations of myDCs and pcDCs were isolated from the blood of antiretroviral treatment-naive patients and assessed for the level of HIV provirus by polymerase chain reaction (PCR). We show that both populations are targets for HIV-1 infection as indicated by the presence of provirus in 12 of 14 pcDC and 13 of 14 myDC samples tested. A proportion of this provirus is integrated in myDCs. The ability of both myDCs and pcDCs from HIV-1-infected patients to stimulate allogeneic T-lymphocyte proliferation in a 6-day mixed leukocyte reaction was severely impaired, but was not mediated by secondary infection of T lymphocytes. Thus, in addition to depletion, both myeloid and plasmacytoid DCs are infected and show impaired functional capacity. These findings suggest that infection, depletion, and dysfunction of dendritic cells may contribute to the immunosuppression associated with HIV-1 disease. PMID- 12576312 TI - A predominant role for cAMP-dependent protein kinase in the cGMP-induced phosphorylation of vasodilator-stimulated phosphoprotein and platelet inhibition in humans. AB - The vasodilator-stimulated phosphoprotein (VASP) plays an important role in cGMP induced platelet inhibition. Since VASP is an in vitro substrate for cGMP dependent protein kinase (PKG), it has been presumed that VASP phosphorylation induced by cGMP is mediated by PKG. Here we show that, in human platelets, phosphorylation of VASP at Ser239 induced by either cGMP analogs or nitric oxide (NO) donor glyco-SNAP1 is inhibited by PKA inhibitors KT5720, PKI, Rp-Br-cAMPS, and H89, but not by PKG inhibitors KT5823 or Rp-pCPT-cGMPS. Unlike human platelets, cGMP analog-induced phosphorylation of VASP in mouse platelets is inhibited by both PKG and PKA inhibitors. Ineffectiveness of PKG inhibitors in inhibiting VASP phosphorylation in human platelets is not due to an inability to inhibit PKG, as these PKG inhibitors but not PKA inhibitors inhibit a different cGMP-induced intracellular signaling event: phosphorylation of extracellular signal-responsive kinase. Furthermore, PKA inhibitors reverse cGMP-induced inhibition of thrombin-induced platelet aggregation, whereas PKG inhibitors further enhance the inhibitory effect of cGMP analogs. Thus, PKA plays a predominant role in the cGMP-induced phosphorylation of VASP and platelet inhibition in human platelets. PMID- 12576313 TI - TCL1 expression in plasmacytoid dendritic cells (DC2s) and the related CD4+ CD56+ blastic tumors of skin. AB - Initially considered to be of natural killer (NK)-cell origin, CD4+ CD56+ blastic tumors (BTs) of skin have recently been proposed to be of dendritic cell lineage. We have previously described BTs with transformation to myelomonocytic leukemia. Here we report expression of the lymphoid proto-oncogene TCL1 in 10 (83%) of 12 BTs and in lymph node plasmacytoid dendritic cells (DC2s). TCL1 was also expressed in myelomonocytic blasts of 3 transformed BT cases but not in true NK cell tumors (n = 18), de novo acute myelomonocytic leukemias (1 of 14, 7%), or mature T-cell malignancies (1 of 112, < 1%), with the exception of T prolymphocytic leukemia (T-PLL). All BT cases were also positive for the DC2 associated marker CD123. These results further support derivation of BTs from DC2s, and demonstrate that TCL1 expression in this tumor is common to the immature blastoid, lymphoid-appearing, and subsequent myelomonocytic phases of this disease. PMID- 12576314 TI - Characterization of molecular defects of Fitzgerald trait and another novel high molecular-weight kininogen-deficient patient: insights into structural requirements for kininogen expression. AB - A 6-year-old male with vertebral-basilar artery thrombosis was recognized to have high-molecular-weight kininogen (HK) deficiency. The propositus had no HK procoagulant activity and antigen (< 1%). Using monoclonal antibodies (Mabs) to kininogen domain 3, the propositus, family members, and Fitzgerald plasma were determined to have detectable low-molecular-weight kininogen. Mabs to HK domains 5 and 6 do not detect HK antigen in the propositus' plasma. The propositus has a single base pair (bp) deletion in cDNA position 1492 of exon 10 affecting amino acid 480 of the mature protein and resulting in a frameshift and a premature stop codon at position 1597 (amino acid 532). Unexpectedly, Mabs to the heavy chain and domain 5 of HK detect a 92-kDa form of HK in Fitzgerald plasma, the first HK deficient plasma. The 92-kDa Fitzgerald HK has amino acid residues through 502, corresponding to domains 1 through 5, but lacks epitopes of domain 6 (positions 543 to 595). Fitzgerald DNA has a normal exon 10, but a 17-bp mutation in intron 9. These combined results indicate that mutations in the kininogen gene may differentially affect biosynthesis, processing, and/or secretion of HK. PMID- 12576315 TI - Inhibition of coagulation, fibrinolysis, and endothelial cell activation by a p38 mitogen-activated protein kinase inhibitor during human endotoxemia. AB - P38 mitogen-activated protein kinase (MAPK) is an important component of intracellular signaling cascades that initiate various inflammatory cellular responses. To determine the role of p38 MAPK in the procoagulant response to lipopolysaccharide (LPS), 24 healthy subjects were exposed to an intravenous dose of LPS (4 ng/kg), preceded 3 hours earlier by orally administered 600 or 50 mg BIRB 796 BS (a specific p38 MAPK inhibitor), or placebo. The 600-mg dose of BIRB 796 BS strongly inhibited LPS-induced coagulation activation, as measured by plasma concentrations of the prothrombin fragment F1 + 2. BIRB 796 BS also dose dependently attenuated the activation and subsequent inhibition of the fibrinolytic system (plasma tissue-type plasminogen activator, plasmin-alpha2 antiplasmin complexes, and plasminogen activator inhibitor type 1) and endothelial cell activation (plasma soluble E-selectin and von Willebrand factor). Activation of p38 MAPK plays an important role in the procoagulant and endothelial cell response after in vivo exposure to LPS. PMID- 12576316 TI - Rituximab plus CHOP (R-CHOP) overcomes bcl-2--associated resistance to chemotherapy in elderly patients with diffuse large B-cell lymphoma (DLBCL). AB - In diffuse large B-cell lymphoma (DLBCL), the combination of rituximab and CHOP (cyclophosphamide, doxorubicine, vincristine, prednisone; R-CHOP) has been shown to be more effective than CHOP for the treatment of elderly patients. Bcl-2 protein expression has been associated with poor prognosis in patients with DLBCL. To establish whether or not rituximab reduces bcl-2-associated treatment failure, we studied bcl-2 protein expression and clinical outcome in patients included in the Groupe d'Etude des Lymphomes de l'Adulte LNH-98-5 trial. Patients between 60 and 80 years of age were randomized to receive 8 cycles of either CHOP or R-CHOP every 3 weeks. Of the 399 patients included, 292 with histologically proven DLBCL had material available for bcl-2 study. Tumors were considered positive when at least 50% of tumor cells expressed bcl-2 protein. There were 193 (66%) bcl-2+ patients and 99 (34%) bcl-2- patients. The response rates for R-CHOP and CHOP were, respectively, 78% and 60% (P =.01) in bcl-2+ patients and 76% and 73% (P =.7) in bcl-2- patients. At a median of 2 years of follow-up, R-CHOP was significantly associated with a better overall survival than CHOP in bcl-2+ patients (67% +/- 9% versus 48% +/- 11%, P =.004). In bcl-2- patients there was no statistically significant difference (72% +/- 12% versus 67% +/- 14%, P =.6). In addition, R-CHOP was associated with significantly better event-free survival than CHOP in bcl-2+ patients (58% +/- 10% versus 32% +/- 10%, P <.001) but not in bcl-2- patients (60% +/- 13% versus 40% +/- 15%, P =.13). Multivariate analysis confirmed the significant benefit for survival and event-free survival of R-CHOP in bcl-2+ patients. These results suggest that rituximab is able to prevent chemotherapy failure in patients with bcl-2 protein overexpression. PMID- 12576317 TI - Proliferation and differentiation potential of human CD8+ memory T-cell subsets in response to antigen or homeostatic cytokines. AB - Four human CD8+ T-cell subsets, naive (CCR7+CD45RA+), central memory (TCM, CCR7+CD45RA-), effector memory (TEM, CCR7-CD45RA-), and CD45RA+ effector memory cells (TEMRA, CCR7-CD45RA+) were compared for their capacity to proliferate and differentiate in response to antigen or homeostatic cytokines. Cytokine responsiveness and interleukin-15 receptor expression were low in naive T cells and progressively increased from TCM to TEM and TEMRA. In contrast, the capacity to accumulate in response to T-cell receptor (TCR) or cytokine stimulation showed a reciprocal pattern and was associated with resistance to cell death and Bcl-2 expression. Whereas all TCR-stimulated cells acquired a CD45RA-CCR7- phenotype, cytokine-stimulated cells maintained their phenotype with the exception of TCM cells, which expressed CCR7, CD45RA, and perforin in various combinations. Single CD8+ TCM cells, but not TEM cells, could be expanded with cytokines, and the obtained clones displayed several distinct phenotypes, suggesting that TCM cells are heterogeneous. Consistently, CCR4 expression in the CD8+ TCM pool discriminated CCR4+ type 2 polarized cells (Tc2) and CCR4-CTL precursors. Finally, ex vivo bromodeoxyuridine (BrdU) incorporation experiments revealed that memory subsets have different in vivo proliferation rates, with CCR4-TCM having the highest turnover and TEMRA the lowest. These results show that human CD8+ memory T-cell subsets have different proliferation and differentiation potentials in vitro and in vivo. Furthermore, they suggest that TEMRA cells are generated from a TCM subset upon homeostatic proliferation in the absence of antigen. PMID- 12576318 TI - Several Bcr-Abl kinase domain mutants associated with imatinib mesylate resistance remain sensitive to imatinib. AB - Imatinib mesylate is a selective Bcr-Abl kinase inhibitor, effective in the treatment of chronic myelogenous leukemia. Most patients in chronic phase maintain durable responses; however, many in blast crisis fail to respond, or relapse quickly. Kinase domain mutations are the most commonly identified mechanism associated with relapse. Many of these mutations decrease the sensitivity of the Abl kinase to imatinib, thus accounting for resistance to imatinib. The role of other mutations in the emergence of resistance has not been established. Using biochemical and cellular assays, we analyzed the sensitivity of several mutants (Met244Val, Phe311Leu, Phe317Leu, Glu355Gly, Phe359Val, Val379Ile, Leu387Met, and His396Pro/Arg) to imatinib mesylate to better understand their role in mediating resistance. While some Abl mutations lead to imatinib resistance, many others are significantly, and some fully, inhibited. This study highlights the need for biochemical and biologic characterization, before a resistant phenotype can be ascribed to a mutant. PMID- 12576319 TI - ADAMTS13 gene mutation in congenital thrombotic thrombocytopenic purpura with previously reported normal VWF cleaving protease activity. AB - Deficiency of von Willebrand factor (VWF) cleaving protease ADAMTS13 is associated with the development of thrombotic thrombocytopenic purpura (TTP). A case of congenital TTP that was previously reported to have normal ADAMTS13 activity was analyzed at the molecular level. Reanalysis of plasma VWF cleaving protease activity using a different assay revealed that the patient had less than 0.1 U/L ADAMTS13 protease activity, while the parents were both partially deficient. Sequence analysis of DNA amplified by polymerase chain reaction showed that the patient was homozygous for a novel TT deletion in exon 15 of the ADAMTS13 gene resulting in a frameshift, while both parents were heterozygous for the same mutation. Taken together with other recent reports, all the cases of hereditary TTP studied by DNA sequence analysis to date appear to be due to mutations within the ADAMTS13 gene. PMID- 12576320 TI - Cell surface peptidase CD26/DPPIV mediates G-CSF mobilization of mouse progenitor cells. AB - CXC ligand 12 (CXCL12; also known as stromal cell-derived factor 1alpha/SDF 1alpha) chemoattracts hematopoietic stem and progenitor cells (HSCs/HPCs) and is thought to play a crucial role in the mobilization of HSCs/HPCs from the bone marrow. CD26 (dipeptidylpeptidase IV [DPPIV]) is a membrane-bound extracellular peptidase that cleaves dipeptides from the N-terminus of polypeptide chains. CD26 has the ability to cleave CXCL12 at its position-2 proline. We found by flow cytometry that CD26 is expressed on a subpopulation of normal Sca-1+c-kit+lin- hematopoietic cells isolated from mouse bone marrow, as well as Sca-1+c-kit-lin- cells, and that these cells possess CD26 peptidase activity. To test the functional role of CD26 in CXCL12-mediated normal HSC/HPC migration, chemotaxis assays were performed. The CD26 truncated CXCL12(3-68) showed an inability to induce the migration of sorted Sca-1+c-kit+lin- or Sca-1+c-kit-lin- mouse marrow cells compared with the normal CXCL12. In addition, CXCL12(3-68) acts as an antagonist, resulting in the reduction of migratory response to normal CXCL12. Treatment of Sca-1+c-kit+lin- mouse marrow cells, and myeloid progenitors within this population, or Sca-1+c-kit-lin- cells with a specific CD26 inhibitor, enhanced the migratory response of these cells to CXCL12. Finally, to test for potential in vivo relevance of these in vitro observations, mice were treated with CD26 inhibitors during granulocyte colony-stimulating factor (G-CSF)-induced mobilization. This treatment resulted in a reduction in the number of progenitor cells in the periphery as compared with the G-CSF regimen alone. This suggests that a mechanism of action of G-CSF mobilization involves CD26. PMID- 12576321 TI - Cappuccino, a mouse model of Hermansky-Pudlak syndrome, encodes a novel protein that is part of the pallidin-muted complex (BLOC-1). AB - Hermansky-Pudlak syndrome (HPS) is a disorder of organelle biogenesis affecting 3 related organelles-melanosomes, platelet dense bodies, and lysosomes. Four genes causing HPS in humans (HPS1-HPS4) are known, and at least 15 nonallelic mutations cause HPS in the mouse. Where their functions are known, the HPS-associated proteins are involved in some aspect of intracellular vesicular trafficking, that is, protein sorting and vesicle docking and fusion. Biochemical and genetic evidence indicates that the HPS-associated genes encode components of at least 3 distinct protein complexes: the adaptor complex AP-3; the HPS1/HPS4 complex; and BLOC-1 (biogenesis of lysosome-related organelles complex-1), consisting of the proteins encoded at 2 mouse HPS loci, pallid (pa) and muted (mu), and at least 3 other unidentified proteins. Here, we report the cloning of the mouse HPS mutation cappuccino (cno). We show that the wild-type cno gene encodes a novel, ubiquitously expressed cytoplasmic protein that coassembles with pallidin and the muted protein in the BLOC-1 complex. Further, we identify a frameshift mutation in mutant cno/cno mice. The C-terminal 81 amino acids are replaced with 72 different amino acids in the mutant CNO protein, and its ability to interact in BLOC-1 is abolished. We performed mutation screening of patients with HPS and failed to identify any CNO defects. Notably, although defects in components of the HPS1/HPS4 and the AP-3 complexes are associated with HPS in humans, no defects in the known components of BLOC-1 have been identified in 142 patients with HPS screened to date, suggesting that BLOC-1 function may be critical in humans. PMID- 12576322 TI - Clinical and biologic implications of recurrent genomic aberrations in myeloma. AB - Nonrandom recurrent chromosomal abnormalities are ubiquitous in multiple myeloma (MM) and include, among others, translocations of the immunoglobulin heavy chain locus (IgH). IgH translocations in MM result in the up-regulation of oncogenes, and include more commonly t(11;14)(q13;q32), t(4;14)(p16;q32), and t(14;16)(q32;q23). Based on the recurrent nature of these translocations and their finding since the early stages of the plasma cell (PC) disorders, we hypothesized that they would confer biologic and clinical variability. In addition, deletions of 13q14 and 17p13 have also been associated with a shortened survival. We used cytoplasmic Ig-enhanced interphase fluorescent in situ hybridization to detect deletions (13q14 and 17p13.1), and translocations involving IgH in 351 patients treated with conventional chemotherapy entered into the Eastern Cooperative Oncology Group clinical trial E9486/9487. Translocations were frequently unbalanced with loss of one of the derivative chromosomes. The presence of t(4; 14)(p16;q32) (n = 42; 26 vs 45 months, P <.001), t(14;16)(q32;q23) (n = 15; 16 vs 41 months, P =.003), - 17p13 (n = 37; 23 vs 44 months, P =.005), and - 13q14 (n = 176; 35 vs 51 months, P =.028) were associated with shorter survival. A stratification of patients into 3 distinct categories allowed for prognostication: poor prognosis group (t(4;14)(p16;q32), t(14; 16)(q32;q23), and - 17p13), intermediate prognosis (- 13q14), and good prognosis group (all others), with median survivals of 24.7, 42.3, and 50.5 months, respectively (P <.001). This molecular cytogenetic classification identifies patients into poor, intermediate, and good risk categories. More importantly it provides further compelling evidence that MM is composed of subgroups of patients categorized according to their underlying genomic aberrations. PMID- 12576323 TI - Jak1 deficiency leads to enhanced Abelson-induced B-cell tumor formation. AB - The Janus kinase Jak1 has been implicated in tumor formation by the Abelson oncogene. In this study we show that loss of Jak1 does not affect in vitro transformation by v-abl as defined by the ability to induce cytokine-independent B-cell colony formation or establishment of B-cell lines. However, Jak1 deficient, v-abl-transformed cell lines were more tumorgenic than wild-type cells when transplanted subcutaneously into severe combined immunodeficient (SCID) mice or injected intravenously into nude mice. Jak1 deficiency was associated with a loss in the ability of interferon-gamma (IFN-gamma)to induce growth arrest and/or apoptosis of v-abl-transformed pre-B cells or tumor growth in SCID mice. Moreover, IFN-gamma mRNA could be detected in growing tumors, and tumor cells explanted from SCID mice had lost the ability to respond to IFN-gamma in 9 of 20 cases, whereas the response to interferon-alpha (IFN-alpha) remained intact. Importantly, a similar increase in tumorgenicity was observed when IFN-gamma deficient cells were injected into SCID mice, identifying the tumor cell itself as the main source of IFN-gamma. These findings demonstrate that Jak1, rather than promoting tumorgenesis as previously proposed, is critical in mediating an intrinsic IFN-gamma-dependent tumor surveillance. PMID- 12576324 TI - CD11c gene expression in hairy cell leukemia is dependent upon activation of the proto-oncogenes ras and junD. AB - Hairy cell leukemia (HCL) is a chronic lymphoproliferative disease, the cause of which is unknown. Diagnostic of HCL is abnormal expression of the gene that encodes the beta2 integrin CD11c. In order to determine the cause of CD11c gene expression in HCL the CD11c gene promoter was characterized. Transfection of the CD11c promoter linked to a luciferase reporter gene indicated that it is sufficient to direct expression in hairy cells. Mutation analysis demonstrated that of predominant importance to the activity of the CD11c promoter is its interaction with the activator protein-1 (AP-1) family of transcription factors. Comparison of nuclear extracts prepared from hairy cells with those prepared from other cell types indicated that hairy cells exhibit abnormal constitutive expression of an AP-1 complex containing JunD. Functional inhibition of AP-1 expressed by hairy cells reduced CD11c promoter activity by 80%. Inhibition of Ras, which represents an upstream activator of AP-1, also significantly inhibited the CD11c promoter. Furthermore, in the hairy cell line EH, inhibition of Ras signaling through mitogen-activated protein kinase/extracellular signal-regulated kinase kinases 1 and 2 (MEK1/2) reduced not only CD11c promoter activity but also reduced both CD11c surface expression and proliferation. Expression in nonhairy cells of a dominant-positive Ras mutant activated the CD11c promoter to levels equivalent to those in hairy cells. Together, these data indicate that the abnormal expression of the CD11c gene characteristic of HCL is dependent upon activation of the proto-oncogenes ras and junD. PMID- 12576325 TI - T-cell epitopes within the complementarity-determining and framework regions of the tumor-derived immunoglobulin heavy chain in multiple myeloma. AB - The idiotypic structure of the monoclonal immunoglobulin (Ig) in multiple myeloma (MM) might be regarded as a tumor-specific antigen. The present study was designed to identify T-cell epitopes of the variable region of the Ig heavy chain (VH) in MM (n = 5) using bioinformatics and analyze the presence of naturally occurring T cells against idiotype-derived peptides. A large number of human leukocyte-antigen (HLA)-binding (class I and II) peptides were identified. The frequency of predicted epitopes depended on the database used: 245 in bioinformatics and molecular analysis section (BIMAS) and 601 in SYFPEITHI. Most of the peptides displayed a binding half-life or score in the low or intermediate affinity range. The majority of the predicted peptides were complementarity determining region (CDR)-rather than framework region (FR)-derived (52%-60% vs 40%-48%, respectively). Most of the predicted peptides were confined to the CDR2 FR3-CDR3 "geographic" region of the Ig-VH region (70%), and significantly fewer peptides were found within the flanking (FR1-CDR1-FR2 and FR4) regions (P <.01). There were 8- to 10-amino acid (aa) long peptides corresponding to the CDRs and fitting to the actual HLA-A/B haplotypes that spontaneously recognized, albeit with a low magnitude, type I T cells (interferon gamma), indicating an ongoing major histocompatibility complex (MHC) class I-restricted T-cell response. Most of those peptides had a low binding half-life (BIMAS) and a low/intermediate score (SYFPEITHI). Furthermore, 15- to 20-aa long CDR1-3-derived peptides also spontaneously recognized type I T cells, indicating the presence of MHC class II restricted T cells as well. This study demonstrates that a large number of HLA binding idiotypic peptides can be identified in patients with MM. Such peptides may spontaneously induce a type I MHC class I- as well as class II-restricted memory T-cell response. PMID- 12576326 TI - Nonmyeloablative conditioning is sufficient to allow engraftment of EGFP expressing bone marrow and subsequent acceptance of EGFP-transgenic skin grafts in mice. AB - Immunologic reactions against gene therapy products may prove to be a frequent problem in clinical gene therapy protocols. Enhanced green fluorescence protein (EGFP) is commonly used as a marker in gene transfer protocols, and immune responses against EGFP-expressing cells have been documented. The present study was designed to investigate the effect of a pharmacologic, nonmyeloablative, conditioning regimen on the development of EGFP+ donor/recipient mixed bone marrow chimerism and ensuing tolerance to EGFP-expressing transplants. To this end, C57BL/6J (B6) mice were treated with soluble formulations of either busulfan (Busulfex) or the closely related compound treosulfan, followed by transplantation of bone marrow cells from EGFP-transgenic (B6-EGFP.Tg) donor mice. Such conditioning regimens resulted in long-term persistence of donor EGFP+ cells among various hematopoietic lineages from blood, bone marrow, and thymus. Stable hematopoietic chimeras transplanted at 10 to 17 weeks after bone marrow transplantation (BMT) with B6-EGFP.Tg skin grafts all accepted their transplants, whereas non-EGFP chimeric B6 control animals were able to mount rejection of the EGFP+ B6 skin grafts. Control third-party grafts from major histocompatibility complex (MHC)-mismatched mice were rejected within 20 days, indicating that acceptance of EGFP-expressing skin grafts was the result of specific immune tolerance induction by the transplantation of EGFP-transgenic bone marrow. Long term tolerance to EGFP in chimeric recipients was confirmed by the absence of anti-EGFP-reactive T cells and antibodies. These results broaden the therapeutic potential for using hematopoietic molecular chimerism in nonmyeloablated recipients as a means of preventing rejection of genetically modified cells. PMID- 12576327 TI - Idiotype vaccination in multiple myeloma induced a reduction of circulating clonal tumor B cells. AB - Myeloma cells express the idiotype (Id)-specific antigen that may be targeted by Id vaccination. Six patients with stage I IgG myeloma were immunized with the autologous purified M component together with the adjuvant cytokines interleukin 12 (IL-12) alone or in combination with granulocyte-macrophage colony-stimulating factor (GM-CSF). The effect of Id vaccination on circulating clonal tumor B cells was monitored by a real-time allele-specific oligonucleotide polymerase chain reaction method. No other treatment was given. Reduction of blood tumor mass was observed in 4 of 6 patients, with one patient achieving a complete molecular remission in blood. In 3 of these 4 patients an Id-specific T-cell response was induced. In the remaining 2 patients with an unchanged level of blood tumor cells, one patient mounted a T-cell response, whereas the other did not. No significant change in the serum M protein level was noted. Id vaccination may target clonal B cells, suggesting that this strategy might be conducive to achieving tumor control. The clinical significance of these findings remains to be established. PMID- 12576328 TI - Differential response of human acute myeloid leukemia cells to gemtuzumab ozogamicin in vitro: role of Chk1 and Chk2 phosphorylation and caspase 3. AB - Gemtuzumab ozogamicin (GO) is a humanized anti-CD33 antibody conjugated to the anticancer agent calicheamicin, approved for the treatment of CD33+-relapsed acute myeloid leukemia. We have investigated the effects of GO on 4 human myeloid leukemia lines of different French-American-British (FAB) types (KG-1, THP-1, HL 60, and NB-4), observing 3 different types of response. Exposure to GO (10-1000 ng/mL) induced G2 arrest (up to 80% of the cells) followed by apoptosis (45% of the cells) in HL-60 and NB-4 cells. By contrast, in THP-1 cells we observed a strong G2 arrest (up to 75% of the cells) with little apoptosis. Finally, the KG 1 line was completely resistant to the same concentrations of GO. These different responses did not correlate with the levels of expression of either CD33 or multiple-drug resistance proteins, although the higher cyclosporin A (CsA) inhibitable efflux activity of KG-1 cells may play a role in the resistance of this line to the drug. We could show that Chk1 and Chk2 phosphorylation, but not p53 or p21 expression, correlated with G2 arrest, implicating the ataxia telangiectasia mutated/ataxia-telangiectasia related (ATM/ATR)-Chk1/Chk2 pathway in the cell cycle response to GO. However, apoptosis was associated with caspase 3 activation. Freshly isolated acute myeloid leukemia (AML) cells showed patterns of response to GO in vitro similar to those observed with the cell lines, including phosphorylation of Chk2 and caspase 3 activation. Our results suggest that the different molecular pathways induced by the drug in vitro may reflect, at least in part, the variable response to GO obtained in vivo. PMID- 12576329 TI - Oxidized phospholipids in oxidized low-density lipoprotein down-regulate thrombomodulin transcription in vascular endothelial cells through a decrease in the binding of RARbeta-RXRalpha heterodimers and Sp1 and Sp3 to their binding sequences in the TM promoter. AB - The present work investigated the mechanism for down-regulation of thrombomodulin (TM), an anticoagulant glycoprotein, on cultured umbilical vein endothelial cells (HUVECs) exposed to lipid extracts from oxidized low-density lipoprotein (ox LDL). HUVECs exposed to phospholipid extracts, but not to free cholesterol, triglyceride, or cholesterol ester, isolated from ox-LDL reduced TM mRNA levels to nearly the same extent as native ox-LDL. Oxidized 1-palmitoyl-2-arachidonyl-sn glycero-3-phosphocholine (ox-PAPC), but not native PAPC or a reduced form of ox PAPC, markedly decreased TM mRNA levels. The apparent half-life (t 1/2 = 2.7 hours) of TM mRNA in control cells was not significantly different from that in cells exposed to ox-LDL or ox-PAPC. TM mRNA levels were regulated by transcriptional activation via a retinoid receptor beta (RARbeta). The binding activities of nuclear proteins from HUVECs treated with ox-LDL or ox-PAPC to the DR4 or stimulatory protein 1 (Sp1) sequence in the TM promoter were significantly reduced with decreased expression of RARbeta, retinoid X receptor alpha (RXRalpha), Sp1, and Sp3 in the nuclei. The promoter activity in HUVECs transfected with a reporter plasmid expressing the TM promoter with targeted deletions in the DR4 and Sp1 binding elements was decreased to about 20% of that with the wild-type construct. Treatment of the cells with ox-PAPC had no additional effect on the promoter activity. These results suggest that oxidized phospholipids in ox-LDL inhibit transcription of the TM gene in HUVECs by inhibiting the binding of RARbeta-RXRalpha heterodimer and Sp, including Sp1 and Sp3, to the DR4 element and Sp1 binding element, respectively, in the TM promoter with reduced expression of RARbeta, RXRalpha, and Sp1 and Sp3 in the nuclei. PMID- 12576330 TI - Survivin is a shared tumor-associated antigen expressed in a broad variety of malignancies and recognized by specific cytotoxic T cells. AB - Survivin, a member of the inhibitor of apoptosis protein family, is expressed in almost all types of malignancies, making this protein a useful tool for the development of broadly applicable vaccination therapies. We used a recently identified HLA-A2 binding peptide and dendritic cells (DCs) from healthy donors to induce survivin-specific cytotoxic T lymphocytes (CTLs) in vitro. These T cells efficiently lysed target cells pulsed with the cognate peptide. Furthermore, survivin-specific CTLs recognized HLA-A2-matched tumor cell lines and primary malignant cells from patients with leukemia in an antigen-specific and HLA-restricted manner as demonstrated with the use of cold target inhibition assays and blocking antibodies. To validate the immunogenicity of survivin we performed the experiments in an autologous setting and used monocyte-derived DCs as targets. Interestingly, we found that DCs up-regulate survivin expression on stimulation with tumor necrosis factor alpha (TNF-alpha). However, these mature DCs were not recognized by survivin-specific CTLs, whereas they lysed autologous mature DCs pulsed with the antigenic peptide or transfected with whole tumor RNA purified from a survivin-expressing cell line. To further analyze the possible use of survivin-specific CTLs in cancer therapies, we induced survivin-specific CTLs using peripheral blood mononuclear cells (PBMNCs) and DCs from a patient with chronic lymphocytic leukemia (CLL). The in vitro-generated T cells efficiently recognized autologous malignant CLL cells, whereas they spared autologous-purified nonmalignant B cells or DCs. Our results demonstrate that survivin epitopes are presented on a broad variety of malignancies and can be applied in vaccination therapies. PMID- 12576331 TI - Regulation of T-cell receptor D beta 1 promoter by KLF5 through reiterated GC rich motifs. AB - Rearrangement of T-cell receptor (TCR) and immunoglobulin genes by a common V(D)J recombination machinery is regulated by developmentally specific chromatin changes at the target locus, a process associated with transcription. At the TCRbeta locus, the Ebeta enhancer and the Dbeta1 promoter regulate germline transcription originating near the TCR Dbeta1 gene segment. The Dbeta1 promoter contains 3 GC-rich motifs that bind a common set of nuclear proteins from pro-T cell lines. Mutations that diminish the binding of nuclear proteins also diminish the activity of the Dbeta1 promoter in transcriptional reporter assays. Using a yeast one-hybrid approach, 3 Kruppel-like factors-KLF3, KLF5, and KLF6-and a novel zinc finger protein were identified in a thymus library, all of which bound the GC-rich motif in a sequence-specific manner. Of these genes, KLF5 mRNA was expressed in a restricted manner in lymphoid cells and tissues, with highest expression in pro-T-cell lines and Rag-deficient thymocytes. Antibody supershift studies and chromatin immunoprecipitation assay confirmed that KLF5 bound the Dbeta1 promoter. In reporter gene assays, KLF5 but not KLF6 efficiently transactivated the Dbeta1 promoter, whereas a dominant-negative KLF5 construct inhibited reporter expression. These data suggest that reiterated GC motifs contribute to germline TCRbeta transcription through binding of KLF5 and other Kruppel family members and that restricted expression of KLF5 may contribute to lineage-specific regulation of germline TCRbeta transcription. PMID- 12576332 TI - RUNX1 and GATA-1 coexpression and cooperation in megakaryocytic differentiation. AB - Megakaryocytic and erythroid lineages derive from a common bipotential progenitor and share many transcription factors, most prominently factors of the GATA zinc finger family. Little is known about transcription factors unique to the megakaryocytic lineage that might program divergence from the erythroid pathway. To identify such factors, we used the K562 system in which megakaryocyte lineage commitment is dependent on sustained extracellular regulatory kinase (ERK) activation and is inhibited by stromal cell contact. During megakaryocytic induction in this system, the myeloid transcription factor RUNX1 underwent up regulation, dependent on ERK signaling and inhibitable by stromal cell contact. Immunostaining of healthy human bone marrow confirmed a strong expression of RUNX1 and its cofactor, core-binding factor beta (CBFbeta), in megakaryocytes and a minimal expression in erythroblasts. In primary human hematopoietic progenitor cultures, RUNX1 and CBFbeta up-regulation preceded megakaryocytic differentiation, and down-regulation of these factors preceded erythroid differentiation. Functional studies showed cooperation among RUNX1, CBFbeta, and GATA-1 in the activation of a megakaryocytic promoter. By contrast, the RUNX1-ETO leukemic fusion protein potently repressed GATA-1-mediated transactivation. These functional interactions correlated with physical interactions observed between GATA-1 and RUNX1 factors. Enforced RUNX1 expression in K562 cells enhanced the induction of the megakaryocytic integrin proteins alphaIIb and alpha2. These results suggest that RUNX1 may participate in the programming of megakaryocytic lineage commitment through functional and physical interactions with GATA transcription factors. By contrast, RUNX1-ETO inhibition of GATA function may constitute a potential mechanism for the blockade of erythroid and megakaryocytic differentiation seen in leukemias with t(8;21). PMID- 12576333 TI - Evaluation of myocardial iron by magnetic resonance imaging during iron chelation therapy with deferrioxamine: indication of close relation between myocardial iron content and chelatable iron pool. AB - Evaluation of myocardial iron during iron chelation therapy is not feasible by repeated endomyocardial biopsies owing to the heterogeneity of iron distribution and the risk of complications. Recently, we described a noninvasive method based on magnetic resonance imaging. Here, the method was used for repeated estimation of the myocardial iron content during iron chelation with deferrioxamine in 14 adult nonthalassemic patients with transfusional iron overload. We investigated the repeatability of the method and the relationship between the myocardial iron estimates and iron status. The repeatability coefficient (2sD) was 2.8 micromol/g in the controls (day-to-day) and 4.0 micromol/g in the patients (within-day). Myocardial iron estimates were elevated in 10 of all 14 patients at first examination, but normalized in 6 patients after 6 to 18 months of treatment. If liver iron declined below 350 micromol/g all but one of the myocardial iron estimates were normal or nearly normal. At start (R2 = 0.69, P =.0014) and still after 6 months of iron chelation (R2 = 0.76, P =.001), the estimates were significantly and more closely related to the urinary iron excretion than to liver iron or serum ferritin levels. In conclusion, our preliminary data, which may only pertain to patients with acquired anemias, suggest the existence of a critical liver iron concentration, above which elevated myocardial iron is present, but its extent seems related to the size of the chelatable iron pool, as reflected by the urinary iron excretion. This further supports the concept of the labile iron pool as the compartment directly involved in transfusional iron toxicity. PMID- 12576334 TI - Persistence of malignant hematopoietic progenitors in chronic myelogenous leukemia patients in complete cytogenetic remission following imatinib mesylate treatment. AB - The BCR/ABL tyrosine kinase inhibitor imatinib mesylate (Gleevec, STI571; Novartis, Basel, Switzerland) has shown remarkable efficacy in the treatment of chronic myelogenous leukemia (CML), with a high proportion of patients achieving complete cytogenetic responses (CCRs). However, it is not clear whether remissions will be durable and whether imatinib mesylate can eliminate the malignant primitive progenitors in which the disease arises. We investigated whether residual BCR/ABL+ hematopoietic progenitors were present in patients who achieved CCRs with imatinib mesylate treatment. CD34+ progenitor cells were selected from bone marrow mononuclear cells (MNCs) and analyzed for the presence of the BCR/ABL fusion gene by fluorescence in situ hybridization (FISH). CD34+ cells were also plated in committed progenitor (colony-forming cell, or CFC) and primitive progenitor (long-term bone marrow culture-initiating cell, or LTCIC) cultures and resulting colonies analyzed for the presence of BCR/ABL+ cells by FISH. Using these assays, residual BCR/ABL+ progenitors were detected in all patients studied. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated increased levels of BCR/ABL mRNA in CD34+ cells compared with total MNCs. Evaluation of samples collected at different time points demonstrated persistence of BCR/ABL+ progenitors despite continued treatment with imatinib mesylate. Our results indicate that inhibition of BCR/ABL tyrosine kinase activity by imatinib mesylate does not eliminate malignant primitive progenitors in CML patients. Patients in CCR with imatinib mesylate treatment need to be followed carefully to assess for risk of relapse. PMID- 12576335 TI - Hemostasis and coagulation at a hematocrit level of 0.85: functional consequences of erythrocytosis. AB - We have generated a transgenic mouse line that reaches a hematocrit concentration of 0.85 due to constitutive overexpression of human erythropoietin in an oxygen independent manner. Unexpectedly, this excessive erythrocytosis did not lead to thrombembolic complications in all investigated organs at any age. Thus, we investigated the mechanisms preventing thrombembolism in this mouse model. Blood analysis revealed an age-dependent elevation of reticulocyte numbers and a marked thrombocytopenia that matched the reduced megakaryocyte numbers in the bone marrow. However, platelet counts were not different from wild-type controls, when calculations were based on the distribution (eg, plasma) volume, thereby explaining why thrombopoietin levels did not increase in transgenic mice. Nevertheless, bleeding time was significantly increased in transgenic animals. A longitudinal investigation using computerized thromboelastography revealed that thrombus formation was reduced with increasing age from 1 to 8 months in transgenic animals. We observed that increasing erythrocyte concentrations inhibited profoundly and reversibly thrombus formation and prolonged the time of clot development, most likely due to mechanical interference of red blood cells with clot-forming platelets. Transgenic animals showed increased nitric oxide levels in the blood that could inhibit vasoconstriction and platelet activation. Finally, we observed that plasmatic coagulation activity in transgenic animals was significantly decreased. Taken together, our findings suggest that prevention of thrombembolic disease in these erythrocytotic transgenic mice was due to functional consequences inherent to increased erythrocyte concentrations and a reduction of plasmatic coagulation activity, the cause of which remains to be elucidated. PMID- 12576336 TI - Human IL-12(p35) gene activation involves selective remodeling of a single nucleosome within a region of the promoter containing critical Sp1-binding sites. AB - To get insight into the regulation of human interleukin-12 (IL-12) synthesis, we determined the chromatin organization of the IL-12(p35) promoter region. First, we determined positioning of nucleosomes within the IL-12(p35) promoter using the indirect end-labeling technique in the THP-1 monocytic cell line. On stimulation with bacterial lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma), hypersensitivity to digestion with DNase I, micrococcal nuclease, and specific restriction enzymes was detected in the region encompassing nucleotide (nt) -310 to -160, indicating selective inducible chromatin remodeling involving disruption of a single nucleosome (named nuc-2). Using p35 promoter deletion mutants and reporter gene assays, we demonstrated that the -396/-241 region contained critical cis-acting elements. Within this latter region, we characterized physically and functionally 2 Sp1-binding sites, which were acting as key regulatory elements for both basal and LPS/IFN-gamma-inducible p35 gene expression: Sp1#1 lies within the remodeled nuc-2 region and Sp1#2 is located in the nucleosome-free region immediately upstream of nuc-2. Finally, we extended the chromatin structure analysis to dendritic cells (DCs) derived from human monocytes and observed the same nucleosomal organization and remodeling as in the THP-1 cell line. Moreover, we found that in DCs, LPS and IFN-gamma synergized in the induction of nucleosomal remodeling and that chromatin remodeling at the p35 locus immediately preceded IL-12(p35) mRNA synthesis. Taken together, our results demonstrate that IL-12(p35) gene activation in the course of DC maturation involves selective and rapid remodeling of a single positioned nucleosome within a region of the promoter containing critical Sp1-binding sites. PMID- 12576337 TI - Impaired recovery of Epstein-Barr virus (EBV)--specific CD8+ T lymphocytes after partially T-depleted allogeneic stem cell transplantation may identify patients at very high risk for progressive EBV reactivation and lymphoproliferative disease. AB - Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes are considered pivotal to prevent lymphoproliferative disease (LPD) in allogeneic stem cell transplantation (SCT) recipients. We evaluated the recovery of EBV-specific CD8+ T cells after partially T-cell-depleted SCT and studied the interaction between EBV-specific CD8+ T cells, EBV reactivation, and EBV-LPD. EBV-specific CD8+ T cells were enumerated using 12 class I HLA tetramers presenting peptides derived from 7 EBV proteins. Blood samples were taken at regular intervals after SCT in 61 patients, and EBV DNA levels were assessed by real-time polymerase chain reaction. Forty-five patients showed EBV reactivation, including 25 with high level reactivation (ie, more than 1000 genome equivalents [geq] per milliliter). Nine of these 25 patients progressed to EBV-LPD. CD8+ T cells specific for latent or lytic EBV epitopes repopulated the peripheral blood at largely similar rates. In most patients, EBV-specific CD8+ T-cell counts had returned to normal levels within 6 months after SCT. Concurrently, the incidence of EBV reactivations clearly decreased. Patients with insufficient EBV-specific CD8+ T-cell recovery were at high risk for EBV reactivation in the first 6 months after SCT. Failure to detect EBV-specific CD8+ T cells in patients with high-level reactivation was associated with the subsequent development of EBV-LPD (P =.048). Consequently, the earlier defined positive predictive value of approximately 40%, based on high level EBV reactivation only, increased to 100% in patients without detectable EBV specific CD8+ T cells. Thus, impaired recovery of EBV-specific CD8+ T cells in patients with high-level EBV reactivation may identify a subgroup at very high risk for EBV-LPD and supports that EBV-specific CD8+ T cells protect SCT recipients from progressive EBV reactivation and EBV-LPD. PMID- 12576338 TI - Down-regulation of BRCA1 in BCR-ABL-expressing hematopoietic cells. AB - BCR-ABL fusion oncogene is the molecular hallmark of chronic myelogenous leukemia (CML), a condition characterized by a progression from a chronic to acute phase leukemia because of secondary genetic events, the nature of which remains largely unknown. Here, we report that the expression of the p210 BCR-ABL fusion protein leads to a down-regulation of BRCA1 protein, a gene product involved in the maintenance of genome integrity. BRCA1 protein is nearly undetectable in leukemia cells from patients with CML, both during the chronic phase and in blast crisis. Similarly, stable transfection-enforced expression of p210 protein in established hematopoietic cell lines leads to severe BRCA1 depletion. The lack of significant change in BRCA1 mRNA level in cells expressing p210 supports the hypothesis that the regulation of BRCA1 protein level occurs after transcription. It is abolished on exposure of the cells to STI571 and by mutation in the adenosine triphosphate (ATP) pocket of p210 and thus seems to require the tyrosine kinase activity of BCR-ABL. Cell lines expressing high levels of BCR-ABL display an increased rate of sister chromatid exchange and chromosome aberrations after ionizing radiation. These findings reveal a novel link between the oncoprotein BCR-ABL and the tumor suppressor protein BRCA1. PMID- 12576339 TI - Epoprostenol therapy as a bridge to pulmonary thromboendarterectomy for chronic thromboembolic pulmonary hypertension. PMID- 12576340 TI - The role of echocardiography in screening for pulmonary arteriovenous malformations. PMID- 12576341 TI - Pembrey's dream: the time has come for a long-term trial of nocturnal supplemental nasal oxygen to treat central sleep apnea in congestive heart failure. PMID- 12576342 TI - Clinical usefulness and limitations of the 6-minute walk test in patients with cardiovascular or pulmonary disease. PMID- 12576343 TI - Getting a leg up on the postthrombotic syndrome. PMID- 12576344 TI - Is it difficult to diagnose sarcoidosis? PMID- 12576345 TI - Chest ultrasound for "Dummies". PMID- 12576346 TI - One from column A: choosing between CT, positron emission tomography, endoscopic ultrasound with fine-needle aspiration, transbronchial needle aspiration, thoracoscopy, mediastinoscopy, and mediastinotomy for staging lung cancer. PMID- 12576347 TI - The diagnostic dilemma in suspected ventilator-associated pneumonia: one size will never fit all. PMID- 12576349 TI - Prostacyclin therapy before pulmonary thromboendarterectomy in patients with chronic thromboembolic pulmonary hypertension. AB - OBJECTIVES: The continuous IV administration of prostacyclin improves pulmonary hemodynamics and prognosis in patients with primary pulmonary hypertension. We investigated whether the administration of prostacyclin therapy to patients before they undergo pulmonary thromboendarterectomy ameliorates pulmonary hypertension in patients with the most severe form of chronic thromboembolic pulmonary hypertension (CTEPH). METHODS: Of the 33 patients with CTEPH who were candidates for pulmonary thromboendarterectomy, 12 patients with severe pulmonary hypertension (pulmonary vascular resistance, > 1,200 dyne. s. cm(-5)) received IV prostacyclin prior to undergoing pulmonary thromboendarterectomy. Right heart catheterization and plasma brain natriuretic peptide (BNP) measurements were repeated at baseline, immediately before surgery, and 1 month after surgery. RESULTS: During a mean (+/- SEM) follow-up period of 46 +/- 12 days, the IV administration of prostacyclin resulted in a 28% decrease in pulmonary vascular resistance (1,510 +/- 53 to 1,088 +/- 58 dyne. s. cm(-5); p < 0.001) before surgery. Prostacyclin therapy markedly decreased plasma BNP level (547 +/- 112 to 188 +/- 30 pg/mL; p < 0.01), suggesting improvement in right heart failure. Pulmonary thromboendarterectomy caused a further reduction of pulmonary vascular resistance (302 +/- 47 dyne. s. cm(-5)) and plasma BNP levels (60 +/- 11 pg/mL) compared to each preoperative value (p < 0.05). Operative mortality rates were relatively low (8.3%) in patients with the most severe form of CTEPH. CONCLUSION: The IV administration of prostacyclin caused beneficial hemodynamic effects in patients with severe CTEPH and may serve as pretreatment for patients undergoing pulmonary thromboendarterectomy. PMID- 12576350 TI - Hemodynamics and survival in patients with pulmonary arterial hypertension related to systemic sclerosis. AB - STUDY OBJECTIVES: The goal of this study was to determine whether the survival of patients with pulmonary hypertension related to systemic sclerosis (SScPH) was different from that of patients with other forms of pulmonary arterial hypertension. DESIGN: Retrospective cohort study. SETTING: Tertiary care medical center. PATIENTS: Our cohort was composed of 33 patients with pulmonary hypertension that is sporadic, familial, or related to anorexigen use (PPH) and 22 patients with SScPH who underwent initial pulmonary artery catheterization and vasodilator study at our center between January 1997 and June 2001. MEASUREMENTS AND RESULTS: Patients with SScPH had somewhat lower percentage of predicted lung volumes than patients with PPH (total lung capacity, 80% vs 92%; p = 0.06) and had lower percentage of predicted diffusion capacity of the lung for carbon monoxide (42% vs 68%; p = 0.0002). Right atrial pressure, pulmonary artery pressure, and cardiac index were similar between the groups. Patients with SScPH and PPH were treated with usual medical therapies, such as digoxin, warfarin, and continuous IV epoprostenol. Despite these similarities, the risk of death in patients with SScPH was higher than in patients with PPH (unadjusted hazard ratio, 2.9; 95% confidence interval, 1.1 to 7.8; p = 0.03). This increased risk appeared to persist after adjustment for a variety of demographic, hemodynamic, or treatment variables. CONCLUSIONS: Despite having similar hemodynamics, patients with SScPH have a higher risk of death than patients with PPH. Future studies of the mechanism and therapy of pulmonary arterial hypertension should focus on the distinctions between the different forms of this disease. PMID- 12576351 TI - Contrast echocardiography remains positive after treatment of pulmonary arteriovenous malformations. AB - STUDY OBJECTIVES: Pulmonary arteriovenous malformations (PAVMs) in patients with hereditary hemorrhagic telangiectasia (HHT) can cause hemorrhage, stroke, and cerebral abscess. Therapy consists of transcatheter embolotherapy (TCET) to occlude the PAVMs. Contrast transthoracic echocardiography (TTE) can be used to screen for PAVMs, but little is known about the performance of contrast TTE after TCET has been performed. Our objective was to determine the effect of the successful performance of TCET on the performance of contrast TTE, specifically, in what proportion of patients the findings of contrast TTE normalized or remained positive after the performance of TCET. DESIGN: Retrospective chart review. SETTING: HHT clinic at university teaching hospital. PATIENTS: Patients who have undergone TCET for the treatment of PAVMs. INTERVENTIONS: Patients were screened for PAVMs with a chest radiograph (CXR), oxygen shunt test (OST), and contrast TTE. Pulmonary angiography was recommended for patients with any positive findings on a screening test. PAVMs > or = 3 mm were occluded by TCET. Contrast TTE, OST, and CXR were performed approximately 1 month later. The results of contrast TTE before and after patients underwent TCET were compared. MEASUREMENTS AND RESULTS: Thirty-nine patients underwent contrast TTE prior to undergoing TCET, and 29 patients underwent contrast TTE both prior to and after undergoing TCET. In all patients, TTE findings were positive prior to TCET. All PAVMs with feeding vessels > or = 3 mm were successfully occluded based on completion angiography. After TCET, 48% of patients had no detectable residual PAVMs, and the remainder had small (ie, < 3 mm) residual PAVMs. Of the 29 patients, 90% had positive contrast TTE findings after undergoing TCET. In the subset of patients who had no residual PAVMs on the completion angiography, 80% had positive contrast TTE findings after undergoing TCET. CONCLUSIONS: In most patients, contrast TTE findings remain positive after they undergo TCET, even in patients without residual PAVMs seen on angiography. This may reflect residual PAVMs that are too small to visualize using angiography. These findings have important implications for the follow-up and management of HHT patients. PMID- 12576352 TI - Clinical outcome after a negative spiral CT pulmonary angiographic finding in an inpatient population from cardiology and pneumology wards. AB - STUDY OBJECTIVES: The purpose of this study was to assess the clinical follow-up of a negative spiral CT (SCT) angiographic finding after a suspicion of acute pulmonary embolism (PE) in a population of inpatients with cardiac and/or respiratory disease. In this high-risk population, clinical findings suggestive of PE are frequently misleading. DESIGN: One hundred seventy-five consecutive patients hospitalized in cardiac and pneumology wards underwent SCT angiography for suspected PE over a 30-month period. Angiographic findings were positive in one third. For the 117 patients with negative SCT angiographic findings, a clinical follow-up during a minimum of 6 months was assessed, particularly in relation to recurrent thromboembolism, mortality, and cause of death. RESULTS: The mean +/- SD follow-up was 21 +/- 11.5 months, and five patients were unavailable for follow-up. Of the 117 patients with negative findings, 81 patients did not receive anticoagulant therapy and 46 patients received anticoagulation for cardiac disease or deep venous thrombosis. Twenty-two patients died during the follow-up period, 3 of them during the first 3 months following the initial event from an undetermined cause. In patients still alive, a new PE occurred in two cases. Patients with a poor cardiopulmonary reserve did not present any recurrent events. In this population, tests other than imaging (d dimers, cardiac echocardiography, or venous ultrasound) contributed little to eliminate the diagnosis of PE. CONCLUSIONS: Whether or not early deaths are considered or not to be related to a recurrent PE, the rate of recurrence after a negative SCT angiographic finding varied between 1.8% and 4.9%. SCT angiography can be used confidently to rule out significant PE, and may prevent further investigations and unnecessary treatment in an inpatient population with cardiac and/or respiratory diseases. PMID- 12576353 TI - Nasal oxygen and muscle sympathetic nerve activity in heart failure. AB - AIMS: To evaluate the effects of mild hyperoxia on sympathetic activity during quiet breathing in patients with chronic heart failure (CHF) and, hence, to investigate whether tonic activation of excitatory chemoreceptor afferents contributes to the elevated sympathetic activity in these patients. Sympathetic activation in patients with CHF may result in part from increased chemoreflex sensitivity. Previous studies using microneurography did not demonstrate deactivation of the chemoreceptors while the patients were breathing 100% O(2). However, 100% O(2) may decrease cardiac output, thereby offsetting the effects on the chemoreflexes. SETTING: University hospital. PATIENTS AND INTERVENTIONS: Ten patients with moderate-to-severe CHF (mean [+/-SD] age, 53.9 +/- 9.2 years; mean ejection fraction, 21.3 +/- 4.7%) were assigned to breathing 20 min of O(2) as well as room air (3 L/min) applied by nasal prongs. Muscle sympathetic nerve activity (MSNA) was evaluated by microneurography of the peroneal nerve. RESULTS: The application of O(2) resulted in an increase of arterial O(2) saturation but no significant change in MSNA during resting ventilation. Although voluntary apneas were no longer with O(2) (25.3 +/- 5.8 vs 32.6 +/- 8.6 s, respectively; p = 0.014), MSNA during the last 10 s of voluntary apnea was lower while breathing O(2) (63.5 +/- 15.0 vs 59.9 +/- 13.9 bursts per minute, respectively; p = 0.02). CONCLUSIONS: The increased MSNA in the patients studied could not be reduced by mild hyperoxia, suggesting that the tonic activation of chemoreflex afferents is unlikely to contribute to the elevated sympathetic activity. That nasal O(2) reduces MSNA during apnea may explain the beneficial effects of nocturnal O(2) therapy in CHF patients with Cheyne-Stokes respiration. PMID- 12576354 TI - Relation between oscillatory ventilation at rest before cardiopulmonary exercise testing and prognosis in patients with left ventricular dysfunction. AB - BACKGROUND: Although nocturnal Cheyne-Stokes respiration alternating between hyperpnea and hypopnea has been considered a sign of severe heart failure, the clinical status of cardiac patients who exhibit oscillatory ventilation during wakefulness has not been clarified. This study was carried out to determine the relation between oscillatory ventilation during wakefulness and exercise capacity in patients with chronic heart disease. We also evaluated retrospectively whether the presence of oscillatory ventilation influences the long-term prognosis in these patients. METHODS: A total of 164 patients with left ventricular dysfunction performed a symptom-limited incremental exercise test. Respiratory gas exchange was measured on a breath-by-breath basis throughout the test. Oscillatory ventilation was defined when clear ventilatory oscillation of at least two consecutive cycles was identified at rest before exercise testing and the difference between the peak and nadir of oscillating ventilation was > 30% of the mean value of ventilation. RESULTS: Oscillatory ventilation was noted in 45 of 164 cardiac patients (27%), and the magnitude (mean +/- SD) of oscillation in these patients was 45.5 +/- 16.9%. Patients with oscillatory ventilation had significantly lower left ventricular ejection fraction than those without it (40.7 +/- 12.7% vs 44.9 +/- 11.6%, p < 0.05). However, parameters of exercise capacity such as the peak oxygen uptake (O(2)), the slope of the increase in O(2) relative to the increase in work rate (DeltaO(2)/DeltaWR), and the ratio of the increase in ventilation to the increase in carbon dioxide output (DeltaE/DeltaCO(2)) were not significantly different between the two groups. The mortality rate during 1,797 +/- 599 days of follow-up did not differ between the groups (p = 0.65). CONCLUSIONS: Oscillatory ventilation present at rest before cardiopulmonary exercise testing is not significantly related to the peak O(2), DeltaO(2)/DeltaWR, DeltaE/DeltaCO(2), or prognosis in patients with left ventricular dysfunction. PMID- 12576355 TI - Limitations of medical therapy in patients with pure coronary spastic angina. AB - OBJECTIVES: To assess the efficacy of medication for the treatment of pure coronary spastic angina, 71 consecutive patients with this diagnosis who had undergone coronary arteriography in a hospital with a follow-up of at least 2 years were studied. METHODS AND RESULTS: All 71 patients without significant organic stenosis were treated with long-acting calcium antagonists. The disappearance of chest pain attacks while receiving medical therapy was observed in 27 patients (38%), whereas the remaining 44 patients (62%) had chest pain attacks. Of special interest, 30 patients had more than one attack per month irrespective of the administration of calcium antagonists or isosorbide dinitrate. Medical treatment showed a good response in female patients (63% vs 31%, respectively; p < 0.05) and those with ST-segment elevation during selective spasm provocation tests (63% vs 30%, respectively; p < 0.05). In contrast, patients with a longer history of chest pain attacks before hospital admission and those with diffuse spasms (77% vs 34%, respectively; p < 0.01) had poor responses to medical treatment. In this study, neither sudden death nor acute myocardial infarction was observed during the follow-up periods. CONCLUSION: The limitations of medical therapy, including the administration of long-acting calcium antagonists, were observed in 30 of 71 patients (42%) with pure coronary spastic angina. Medical treatment was effective in only 38% of patients with pure coronary spastic angina in Japan. PMID- 12576356 TI - The 6-min walk test: a quick measure of functional status in elderly adults. AB - OBJECTIVES: To determine the correlates of the total 6-min walk distance (6MWD) in a population sample of adults > or = 68 years old. METHODS: The standardized 6 min walk test (6MWT) was administered to the Cardiovascular Health Study cohort during their seventh annual examination. RESULTS: Of the 3,333 participants with a clinic visit, 2,281 subjects (68%) performed the 6MWT. There were no untoward events. The mean 6MWD was 344 m (SD, 88 m). Independent general correlates of a shorter 6MWD in linear regression models in women and men included the following: older age, higher weight, larger waist, weaker grip strength, symptoms of depression, and decreased mental status. Independent disease or risk factor correlates of a shorter 6MWD included the following: a low ankle BP, use of angiotensin-converting enzyme inhibitors, and arthritis in men and women; higher C-reactive protein, diastolic hypertension, and lower FEV(1) in women; and the use of digitalis in men. Approximately 30% of the variance in 6MWD was explained by the linear regression models. Newly described bivariate associations of a shorter 6MWD included impaired activities of daily living; self-reported poor health; less education; nonwhite race; a history of coronary heart disease, transient ischemic attacks, stroke, or diabetes; and higher levels of C-reactive protein, fibrinogen, or WBC count. CONCLUSIONS: Most community-dwelling elderly persons can quickly and safely perform this functional status test in the outpatient clinic setting. The test may be used clinically to measure the impact of multiple comorbidities, including cardiovascular disease, lung disease, arthritis, diabetes, and cognitive dysfunction and depression, on exercise capacity and endurance in older adults. Expected values should be adjusted for the patient's age, gender, height, and weight. PMID- 12576357 TI - Acute effects of exercise in patients with previous deep venous thrombosis: impact of the postthrombotic syndrome. AB - BACKGROUND: The postthrombotic syndrome (PTS) occurs frequently after deep venous thrombosis (DVT) and is believed to worsen with upright posture and physical activity. However, the effects of exercise in patients with previous DVT have not been studied. STUDY OBJECTIVES: To determine whether previous DVT and PTS limit the ability to exercise, and whether exercise increases the severity of venous symptoms and signs. DESIGN AND SETTING: A repeated-measures cohort study that was conducted at a university-affiliated teaching hospital, 1999-2000. PARTICIPANTS: Subjects with a first episode of unilateral DVT at least 1 year earlier were recruited from the Thrombosis Clinic (total, 41 subjects; with PTS, 19 subjects). INTERVENTION: Treadmill exercise session. MEASUREMENTS AND RESULTS: Venous symptoms, calf muscle fatigability, flexibility, and leg volume before and after treadmill exercise were measured and compared. Exercise did not worsen venous symptoms, despite a higher gain in affected leg volume in subjects with PTS vs subjects without PTS (mean difference: affected leg, 53 mL; unaffected leg, -15 mL; p = 0.018). Calf flexibility significantly improved after exercise in subjects with PTS (gastrocnemius: affected-unaffected, PTS vs no PTS + 4.5 degrees, p = 0.0029; soleus: affected-unaffected, PTS vs no PTS + 5.7 degrees, p = 0.0011). CONCLUSIONS: Exercise did not acutely exacerbate symptoms and, in subjects with PTS, resulted in improved flexibility in the affected leg. Our findings suggest that treadmill or similar exercise is unlikely to make symptoms of PTS worse, and may improve flexibility. Further study is indicated to determine whether a regular exercise-training program might have a role in the management of patients with PTS, since, to date, the treatment options for this condition are limited. PMID- 12576358 TI - The diagnostic pathway to sarcoidosis. AB - PURPOSE: To examine the time from the first physician visit to the diagnosis of sarcoidosis. HYPOTHESES: The time required to diagnose sarcoidosis is dependent on the initial symptoms, socioeconomic status, referral to a specialist, race, and severity of pulmonary involvement. METHODS: Patients were recruited from the Case Control Etiology of Sarcoidosis Study (ACCESS) and had biopsy-confirmed sarcoidosis. Subjects were asked to recall the date of onset of symptoms of sarcoidosis, their first physician visit, number of physician visits, and types of physicians seen. RESULTS: One hundred eighty-nine patients were enrolled. The diagnosis of sarcoidosis was made on the first physician visit in only 15.3% of cases. The presence of pulmonary symptoms was associated with prolonged time (> 6 months vs < or = 6 months, p = 0.02) until diagnosis, and the presence of skin symptoms with a shorter time (< or = 6 months vs > 6 months, p = 0.02) until diagnosis. Patients with pulmonary symptoms had more physician visits (mean +/- SEM) until the diagnosis was made compared to those without pulmonary symptoms (4.84 +/- 0.38 visits vs 3.15 +/- 0.24 visits, p = 0.0002). The mean baseline FEV(1) was greater in those diagnosed < or = 6 months from the first physician visit than those diagnosed > 6 months (87.3 +/- 1.52% predicted vs 81.2 +/- 2.5% predicted, p = 0.04). There was a significant delay in diagnosis (> 6 months vs < or = 6 months) from first physician visit with higher Scadding stages (stage 4 vs stage 2, or stage 3 vs stage 0 or 1, p = 0.04). CONCLUSIONS: The diagnosis of sarcoidosis is often delayed and seems to be more a factor of disease presentation than patient or physician characteristics. The presence of pulmonary symptoms or higher radiographic stages is associated with a prolonged time until diagnosis. The presence of skin symptoms is associated with less delay in diagnosis. It is likely that the delay in diagnosis of pulmonary sarcoidosis relates to the fact that pulmonary symptoms and parenchymal involvement are nonspecific and are often regarded as manifestations of other pulmonary diseases. PMID- 12576359 TI - Recovery of cell wall-deficient organisms from blood does not distinguish between patients with sarcoidosis and control subjects. AB - STUDY OBJECTIVE: To determine if cell wall-deficient forms (CWDF) of mycobacteria can be grown in culture of blood from subjects with sarcoidosis. DESIGN: A special multicenter study of sarcoidosis (A Case Control Etiologic Study of Sarcoidosis), supported by the National Heart, Lung, and Blood Institute. PATIENTS AND CONTROL SUBJECTS: PATIENTS AND CONTROL SUBJECTS were recruited at 10 institutions in the United States. Control subjects (controls) were of the same gender and race, and within 5 years of age as matching patients with sarcoidosis (cases). RESULTS: Cultures were incubated from 347 blood specimens (197 cases, 150 controls). Two investigators trained to recognize CWDF mycobacteria examined material obtained from culture tubes after 3 weeks. Structures thought to be CWDF were seen with equal frequency in cases (38%) and controls (41%). Thirty-nine percent of cases and 37% of controls were read as negative for CWDF. CONCLUSION: This study fails to confirm earlier reports that CWDF mycobacteria can be grown from the blood of patients with sarcoidosis, but not from control subjects. PMID- 12576360 TI - Ultrasound-guided thoracentesis: is it a safer method? AB - STUDY OBJECTIVES: The objectives of this study are as follows: (1) to determine the incidence of complications from thoracentesis performed under ultrasound guidance by interventional radiologists in a tertiary referral teaching hospital; (2) to evaluate the incidence of vasovagal events without the use of atropine prior to thoracentesis; and (3) to evaluate patient or radiographic factors that may contribute to, or be predictive of, the development of re-expansion pulmonary edema after ultrasound-guided thoracentesis. DESIGN: Prospective descriptive study. SETTING: Saint Thomas Hospital, a tertiary referral teaching hospital in Nashville, TN. PATIENTS: All patients referred to interventional radiology for diagnostic and/or therapeutic ultrasound-guided thoracentesis between August 1997 and September 2000. RESULTS: A total of 941 thoracenteses in 605 patients were performed during the study period. The following complications were recorded: pain (n = 25; 2.7%), pneumothorax (n = 24; 2.5%), shortness of breath (n = 9; 1.0%), cough (n = 8; 0.8%), vasovagal reaction (n = 6; 0.6%), bleeding (n = 2; 0.2%), hematoma (n = 2; 0.2%), and re-expansion pulmonary edema (n = 2; 0.2%). Eight patients with pneumothorax received tube thoracostomies (0.8%). When > 1,100 mL of fluid were removed, the incidence of pneumothorax requiring tube thoracostomy and pain was increased (p < 0.05). Fifty-seven percent of patients with shortness of breath during the procedure were noted to have pneumothorax on postprocedure radiographs, while 16% of patients with pain were noted to have pneumothorax on postprocedure radiographs. Vasovagal reactions occurred in 0.6% despite no administration of prophylactic atropine. Re-expansion pulmonary edema complicated 2 of 373 thoracenteses (0.5%) in which > 1,000 mL of pleural fluid were removed. CONCLUSIONS: The complication rate with thoracentesis performed by interventional radiologists under ultrasound guidance is lower than that reported for non-image-guided thoracentesis. Premedication with atropine is unnecessary given the low incidence of vasovagal reactions. Re-expansion pulmonary edema is uncommon even when > 1,000 mL of pleural fluid are removed, as long as the procedure is stopped when symptoms develop. PMID- 12576361 TI - Continuous thoracic paravertebral infusion of bupivacaine for pain management in patients with multiple fractured ribs. AB - STUDY OBJECTIVE: To evaluate the efficacy of a continuous thoracic paravertebral infusion of bupivacaine for pain management in patients with unilateral multiple fractured ribs (MFR). DESIGN: Prospective nonrandomized case series. SETTING: Multidisciplinary tertiary hospital. PATIENTS: Fifteen patients with unilateral MFR. INTERVENTIONS: Insertion of a catheter into the thoracic paravertebral space. We administered an initial injection of 0.3 mL/kg (1.5 mg/kg) bupivacaine 0.5% with 1:200,000 epinephrine followed 30 min later by an infusion of bupivacaine 0.25% at 0.1 to 0.2 mL/kg/h for 4 days. MEASUREMENTS AND RESULTS: The following parameters were measured during the initial assessment before thoracic paravertebral block (TPVB), 30 min after the initial injection, and during follow up on day 1 and day 4 after commencing the infusion of bupivacaine: visual analog pain score at rest and during coughing; respiratory rate; arterial oxygen saturation (SaO(2)); bedside spirometry (ie, FVC, FEV(1), FEV(1)/FVC ratio, and peak expiratory flow rate [PEFR]); arterial blood gas measurements; and O(2) index (ie, PaO(2)/fraction of inspired oxygen ratio). There were significant improvements in pain scores (at rest, p = 0.002; during coughing, p = 0.001), respiratory rate (p < 0.0001), FVC (p = 0.007), PEFR (p = 0.01), SaO(2) (p = 0.04), and O(2) index (p = 0.01) 30 min after the initial injection, which were sustained for the 4 days that the thoracic paravertebral infusion was in use (p < 0.05). PaCO(2) did not change significantly after the initial injection, but on day 4 it was significantly lower than the post-TPVB value (p = 0.04). One patient had an inadvertent epidural injection, and another developed transient ipsilateral Horner syndrome with sensory changes in the arm. No patient exhibited clinical signs of inadvertent intravascular injection or local anesthetic toxicity. CONCLUSION: Our results confirmed that continuous thoracic paravertebral infusion of bupivacaine is a simple and effective method of providing continuous pain relief in patients with unilateral MFR. It also produced a sustained improvement in respiratory parameters and oxygenation. PMID- 12576362 TI - Effect of intrapleural streptokinase administration on antistreptokinase antibody level in patients with loculated pleural effusions. AB - BACKGROUND: Streptokinase is widely used IV for the treatment of myocardial infarction and intrapleurally for the treatment of loculated pleural effusions. IV administration of streptokinase is known to cause the production of antistreptokinase antibodies. OBJECTIVE: The aim of this study was to evaluate whether the intrapleural administration of streptokinase results in a similar elevation of the serum antistreptokinase antibody level. METHODS: During 1 year, venous blood samples were taken from 16 consecutive patients (10 men and 6 women; age range, 22 to 60 years) requiring intrapleural streptokinase administration (250,000 IU once a day, for 2 to 6 days). Blood samples were taken before treatment, on day 5, and day 14. Antistreptokinase antibodies were measured using enzyme-linked immunosorbent assay (ELISA) and were expressed in arbitrary ELISA units. Four patients with myocardial infarction treated with IV streptokinase (1,500,000 IU) were included as control subjects for the method. RESULTS: Before treatment, the median antistreptokinase antibody level in patients with loculated pleural effusions was 729 ELISA units (range, 196 to 13,529 ELISA units) and increased to 9,240 ELISA units (range, 1,456 to 77,389 ELISA units) by day 14 (p < 0.0001). In the control group, the median pretreatment level was 119 ELISA units, and by day 14 it had increased to 20,495 ELISA units. Four patients who developed an elevated body temperature after intrapleural administration of streptokinase had a significantly higher pretreatment antistreptokinase antibody level compared to other patients. CONCLUSIONS: The intrapleural administration of streptokinase results in the elevation of the serum antistreptokinase antibody level, which is similar to the case with IV administration. An increased pretreatment antistreptokinase antibody level does not influence the result of intrapleural fibrinolysis but can cause an elevation of body temperature after the administration of streptokinase. PMID- 12576363 TI - Accuracy of pleural puncture sites: a prospective comparison of clinical examination with ultrasound. AB - STUDY OBJECTIVE: To assess the value of chest ultrasonography vs clinical examination for planning of diagnostic pleurocentesis (DPC). DESIGN: Prospective comparative study. SETTING: Pulmonary unit of a tertiary teaching hospital. PATIENTS AND PARTICIPANTS: Sixty-seven consecutive patients referred to 30 physicians of varying degrees of experience for DPC. INTERVENTIONS: Based on clinical data and examination, physicians determined whether and where a DPC should be performed. Selected puncture sites were evaluated with ultrasound and considered accurate when > or = 10 mm fluid perpendicular to the skin were present. MEASUREMENTS AND RESULTS: In 172 of 255 cases (67%), a puncture site was proposed. Twenty-five sites (15%) were found to be inaccurate on ultrasound examination, and a different, accurate site was established in 20 of these cases. Physicians were unable to locate a puncture site in 83 cases (33%). Among these, ultrasound demonstrated an accurate site in 45 cases (54%), while a safe tap was truly impossible in 38 cases (46%). Overall, ultrasound prevented possible accidental organ puncture in 10% of all cases and increased the rate of accurate sites by 26%. The sensitivity and specificity for identifying a proper puncture site with clinical examination compared to ultrasound as the "gold standard" were 76.6% and 60.3% (positive and negative predictive values, 85.5% and 45.8%, respectively). Risk factors associated with inaccurate clinical site selection were as follows: small effusion (p < 0.001), evidence of fluid loculation on chest radiography (p = 0.01; relative risk, 7.8; 95% confidence interval, 1.9 to 32.9), and sharp costodiaphragmatic angle on chest radiography (p < 0.001; relative risk, 7.0; 95% confidence interval, 2.3 to 15.2). Experienced physicians did not perform better than physicians in training. CONCLUSIONS: Puncture site selection with bedside ultrasonography increases the yield of and potentially reduces complication rate in DPC. Physician experience does not predict the accuracy of selected puncture sites. PMID- 12576364 TI - Mediastinal lymph node involvement in potentially resectable lung cancer: comparison of CT, positron emission tomography, and endoscopic ultrasonography with and without fine-needle aspiration. AB - PURPOSE: A prospective comparison of three imaging techniques: thoracic CT, positron emission tomography (PET), and endoscopic ultrasonography (EUS) with fine needle aspiration (FNA), each performed under routine conditions, for the detection of metastatic lymph nodes metastases in patients with lung cancer considered for operative resection. PATIENTS AND METHODS: Following bronchoscopic evaluation, CT, PET, and EUS were performed to evaluate potential mediastinal involvement in 33 consecutive patients with bronchoscopic biopsy/cytology proven (n = 25) or radiologically suspected (n = 8) lung cancer prior to surgery. Surgical histology was used as "gold standard" to confirm the diagnosis of the primary tumor and the mediastinal status in all patients. Histology proved non small cell lung cancer in 30 patients, neuroendocrine tumor in 1 patient, and benign disease in 2 patients. RESULTS: The mean age of the study group was 61.5 years (range, 41 to 80 years; 23 male patients). CT, PET, and EUS detected mediastinal lymph nodes (size, 0.4 to 1.6 cm) in 15, 14, and 27 patients (21 of which were suspected to be malignant on EUS), respectively. With respect to the correct prediction of mediastinal lymph node stage, the sensitivities of CT, PET, and EUS were 57%, 73%, and 94%. Specificities were 74%, 83%, and 71%. Accuracies were 67%, 79%, and 82%. Results of PET could be improved when combined with CT (sensitivity, 81%; specificity, 94%; accuracy, 88%). The specificity of EUS (71%) was improved to 100% by FNA cytology (EUS-guided FNA), which gave a tissue diagnosis including tumor type, without complications. CONCLUSIONS: No single imaging method alone was conclusive in evaluating potential mediastinal involvement in apparently operable lung cancer and routine clinical conditions. A tissue diagnosis is extremely helpful. Because FNA can be performed at the same time as EUS, this combination emerged as the most useful technique in the evaluation of even very small mediastinal metastases of lung cancer. CT seems necessary additionally to evaluate the pretracheal region as well as the rest of the thorax, and PET may be valuable to detect distant metastases. PMID- 12576365 TI - Patient use of the internet for information in a lung cancer clinic. AB - STUDY OBJECTIVES: To determine how frequently patients attending a lung cancer clinic use the Internet for their own health information, to determine whether there are demographic differences between Internet users and nonusers, and to determine how patients compare the quality of Internet information with other sources of lung cancer information. DESIGN: Sequentially administered patient questionnaire. One hundred eighty-four patients were surveyed, and 139 patients (75.5%) completed the questionnaire. SETTING: A multidisciplinary thoracic oncology clinic in a Midwestern University hospital. PATIENTS OR PARTICIPANTS: Patients attending the multidisciplinary thoracic oncology clinic over a 3-month period. MEASUREMENTS AND RESULTS: The Internet was the most commonly used nonphysician source of information among our patients. Sixteen percent of the patients sought information on the Internet, but 60% expressed interest in using the Internet for information. Users were on average of higher income level and educational attainment but did not differ from nonusers by community size. Internet users rated the quality of information available on the Internet of similar quality to information from all sources. CONCLUSIONS: Older patients are increasing using the Internet for self-education in lung cancer. While certain barriers continue to exist, patients from rural areas use the Internet to the same degree, as do patients from urban areas. Patients do, however, overrate the quality of information on the Internet. PMID- 12576366 TI - Endobronchial ultrasound reliably differentiates between airway infiltration and compression by tumor. AB - OBJECTIVE: A frequent problem in patients with intrathoracic malignancies neighboring central airways is the question of whether the airway wall is infiltrated by the tumor or if it is merely compressed. This distinction can often not be made with certainty with the help of chest CT alone, but frequently necessitates surgical biopsy or exploration. We prospectively studied the utility of endobronchial ultrasound (EBUS) in this clinical circumstance. METHODS AND PATIENTS: Between May 1999 and July 2000, 131 consecutive patients with central thoracic malignancies potentially involving the airways were enrolled into the study. Patients underwent chest CT followed by standard bronchoscopy together with EBUS and subsequent surgical evaluation. The bronchoscopists did not know the radiologist's interpretation of the chest CT before performing EBUS. The ability of chest CT and EBUS to distinguish between compression and infiltration was measured against the histologic results. RESULTS: One hundred five patients completed the trial by undergoing surgery. In 81 patients (77%), the CT scan was read as consistent with tumor invasion. EBUS only showed invasion in 49 cases (47%). Histology after surgery revealed a specificity of 100%, a sensitivity of 89%, and an accuracy of 94% for EBUS. Chest CT was far inferior, with a specificity of 28%, a sensitivity of 75%, and an accuracy of 51%. CONCLUSION: We conclude that EBUS is a highly accurate diagnostic tool and superior to chest CT in evaluating the question of airway involvement by central intrathoracic tumors. In the hands of experienced endoscopists, EBUS may become the procedure of choice for this question. PMID- 12576367 TI - Lymph node size and metastatic infiltration in non-small cell lung cancer. AB - BACKGROUND: Preoperative lymph node staging of lung cancer by CT relies on the premise that malignant lymph nodes are larger than benign ones. Lymph nodes > 1 cm in size are regarded as metastatic nodes. The surgical approach and potential application of neoadjuvant therapy regimens are dependent on this evaluation. PATIENTS AND METHODS: In a morphometric study, hilar and mediastinal lymph nodes from 256 patients with non-small cell lung cancer (NSCLC) were analyzed. The lymph nodes were counted, the largest diameter of each lymph node was measured, and each lymph node was analyzed for metastatic involvement by histopathologic examination. The frequency of metastatic involvement was calculated and correlated with lymph node size. Preoperative CT scans of 80 patients were retrospectively analyzed by a staff radiologist. Lymph node size was measured, and lymph nodes were evaluated due to radiologic criteria. The radiologic evaluation was compared to the histopathologic diagnosis. RESULTS: A total of 2,891 lymph nodes were present in the 256 specimens examined for this study. One hundred thirty-nine patients had a pN0 status, whereas 117 patients had lymph nodes that were positive for cancer. Two thousand four hundred eighty-six lymph nodes (86%) were tumor-free, while 405 (14%) showed metastatic involvement on histopathologic examination. The mean (+/- SD) diameter of the nonmetastatic lymph nodes was 7.05 +/- 3.75 mm, whereas infiltrated nodes had a diameter of 10.7 +/- 4.7 mm (p = 0.005). One thousand nine hundred fifty-three of the tumor free lymph nodes (79%) and 170 of the metastatic lymph nodes (44%) were < 10 mm in diameter. Of 139 patients with no metastatic lymph node involvement, 101 (77%) had at least one lymph node that was > 10 mm in diameter. Of 127 patients with metastatic lymph node involvement, 12% had no lymph node that was < 10 mm. The independent radiologic evaluation of the CT scans of 80 patients yielded a sensitivity of 57.1% and a specificity of 80.6%. CONCLUSION: Lymph node size is not a reliable parameter for the evaluation of metastatic involvement in patients with NSCLC. PMID- 12576368 TI - Inspiratory stridor in elite athletes. AB - STUDY OBJECTIVES: Diagnosis and medical intervention for exercise-induced bronchospasm (EIB) are often based on self-reported symptoms, without spirometric confirmation. Inspiratory stridor (IS), a symptom of vocal cord dysfunction (VCD), is frequently mistaken for EIB wheeze. Athletes with exercise IS that spontaneously resolves on activity cessation are suspect for VCD and may not have EIB. This study estimated IS prevalence in elite athletes and determined its relationship to EIB. SUBJECTS/METHODS: Three hundred seventy athletes (174 female and 196 male subjects) provided a medical history, and underwent spirometry before and after exercise challenge. Exercise challenges were conducted in cold, dry ambient conditions. EIB positive (EIB +) was defined as a > or = 10% postexercise fall in FEV(1). Athletes were monitored for IS during exercise; 78.4% of the athletes in this study (n = 290) were tested on multiple occasions. RESULTS: EIB was identified in 30% of 370 athletes tested (58 female and 53 male subjects). IS was observed in 5.1% (18 female and 1 male subjects) during exercise and spontaneously resolved in these subjects within 5 min after exercise cessation. Ten IS-positive (IS +) athletes (52.6%) were EIB +, and 8 of these athletes had a previous EIB diagnosis; however, beta(2)-agonist treatment resolved IS in only 2 subjects. Eight of nine IS +/EIB-negative (EIB -) athletes had a previous EIB diagnosis; seven subjects received beta(2)-agonist treatment with no IS resolution. Resting spirometric measurements did not distinguish IS, but postexercise mid-flow (FEF(50)/FIF(50)) ratio > 1.5 was more frequent (33%, p < 0.05) among IS + athletes. The FEF(50)/FIF(50) ratio was higher for IS +/EIB + athletes than for IS -/EIB + athletes (1.97 +/- 1.69 vs 0.81 +/- 0.39, p < 0.05). The postexercise fall in FVC was greater (p < 0.05) for IS +/EIB - athletes (9.2 +/- 5.0%) than for IS-negative (IS -) /EIB - athletes (5.3 +/- 4.3%). No difference in postexercise FEV(1) was identified between IS + and IS - athletes (within EIB + or EIB - groups). CONCLUSIONS: Five percent of athletes were IS +, with EIB comorbidity observed in 53% of these subjects. Misdiagnosis of IS as EIB is common. The lack of a beta(2)-agonist response in combination with postexercise serial spirometry can be useful in excluding solitary IS and confirming EIB diagnosis. PMID- 12576369 TI - Prolonged airway and systemic inflammatory reactions after smoke inhalation. AB - STUDY OBJECTIVES: Smoke inhalation has a prolonged, negative effect on pulmonary function. The immediate change in the airway after smoke inhalation is an intense inflammatory reaction. Obstructive airway disease commonly occurs several years after smoke inhalation, but few studies have focused on long-term reactions in the airway. This study investigated the long-term effects of smoke inhalation, by examining airway responsiveness, airway inflammation, and systemic effects. DESIGN: Cross-sectional study. PATIENTS: We assessed victims (n = 9) of smoke inhalation 6 months after they were exposed. INTERVENTIONS: We studied the clinical symptoms, laboratory data, and pulmonary functions of the patients. We also performed the nonspecific bronchial challenge test with methacholine on these patients. In some patients, we reviewed pathologic specimens of bronchi and measured cytokines (tumor necrosis factor [TNF]-alpha, interferon [INF]-gamma, and interleukin [IL]-2) in serum and BAL fluid. RESULTS: All the subjects complained of a productive cough, and three subjects had a mild degree of dyspnea on exertion. All but one subject had airway hyperresponsiveness to methacholine. The pulmonary function test results, however, were within normal limits, except for one subject who had a mild obstructive pattern of pulmonary function. Bronchial mucosal biopsy (n = 2) showed inflammatory changes with lymphocyte infiltration. Significantly greater concentrations of TNF-alpha (mean, 1,346.4 pg/mL vs 61.2 pg/mL; p < 0.05) and IFN-gamma (mean, 540.9 pg/mL vs 26.7 pg/mL; p < 0.05) were seen in the serum (n = 4) compared with control subjects. The serum IL-2 level was also increased (mean, 136.8 pg/mL vs undetectable); however, the increase was not significant compared with the control subjects. CONCLUSIONS: These data suggest that inflammatory reactions in the airways and peripheral blood continue for at least 6 months after smoke inhalation. PMID- 12576370 TI - Nebulized 3% hypertonic saline solution treatment in hospitalized infants with viral bronchiolitis. AB - OBJECTIVE: To determine the utility of inhaled hypertonic saline solution to treat infants hospitalized with viral bronchiolitis. DESIGN: Randomized, double blind, controlled trial. Fifty-two hospitalized infants (mean +/- SD age, 2.9 +/- 2.1 months) with viral bronchiolitis received either inhalation of epinephrine, 1.5 mg, in 4 mL of 0.9% saline solution (group 1; n = 25) or inhalation of epinephrine, 1.5 mg, in 4 mL of 3% saline solution (group 2; n = 27). This therapy was repeated three times every hospitalization day until discharge. RESULTS: The percentage improvement in the clinical severity scores after inhalation therapy was not significant in group 1 on the first, second, and third days after hospital admission (3.5%, 2%, and 4%, respectively). In group 2, significant improvement was observed on these days (7.3%, 8.9%, and 10%, respectively; p < 0.001). Also, the improvement in clinical severity scores differed significantly on each of these days between the two groups. Using 3% saline solution decreased the hospitalization stay by 25%: from 4 +/- 1.9 days in group 1 to 3 +/- 1.2 days in group 2 (p < 0.05). CONCLUSIONS: We conclude that in nonasthmatic, nonseverely ill infants hospitalized with viral bronchiolitis, aerosolized 3% saline solution/1.5 mg epinephrine decreases symptoms and length of hospitalization as compared to 0.9% saline solution/1.5 mg epinephrine. PMID- 12576371 TI - Pulmonary infiltrates in patients receiving long-term glucocorticoid treatment: etiology, prognostic factors, and associated inflammatory response. AB - BACKGROUND: Glucocorticoid treatment alters immunoregulatory defense mechanisms and may therefore favor the development of different pulmonary infections. METHODS: The etiology, prognostic factors, and associated inflammatory response of pulmonary infiltrates in 33 patients receiving long-term glucocorticoid treatment (LTGCT) were prospectively evaluated. RESULTS: Aspergillus spp (n = 9, 31%) and Staphylococcus spp (n = 6, 21%) were the most common causative agents. Using different diagnostic techniques, we obtained a specific diagnosis in 28 of 33 episodes (85%) of pulmonary infiltrates. Bronchoscopic techniques provided the diagnosis in 64% of the cases. Crude mortality was 45%. Variables associated with mortality were as follows: age > 64 years, bilateral radiographic involvement, delay in diagnosis, inappropriate empirical treatment, Simplified Acute Physiology Score (SAPS) II > or = 25, and requirement for mechanical ventilation (MV). SAPS II > or = 25 (odds ratio [OR], 16; 95% confidence interval, 1 to 260) and MV requirement (OR, 50; 95% confidence interval, 2 to 360) were also significant on multivariate analysis. Pulmonary infections were associated with an increase in the concentration of relevant inflammatory cytokines such as tumor necrosis factor-alpha and interleukin-6 both in serum and BAL. This local and systemic inflammatory response was attenuated when compared with the response observed in patients with pulmonary infections but without glucocorticoid treatment or receiving glucocorticoids for a short period of time (< 9 days). CONCLUSIONS: Pulmonary infiltrates in patients receiving LTGCT are often caused by fungi and Gram-positive cocci, and are associated with attenuated local and systemic inflammatory response. Although in most cases, sputum cultures and bronchoscopic techniques are diagnostic, the associated mortality is high, particularly in those requiring MV. PMID- 12576372 TI - Warfarin dose reduction vs watchful waiting for mild elevations in the international normalized ratio. AB - BACKGROUND: Whether clinicians should decrease the warfarin dose in response to a mild, asymptomatic elevation in the international normalized ratio (INR) is unknown. OBJECTIVES: The study objectives were as follows: (1) to evaluate the safety of an anticoagulation service (ACS) policy advocating that the warfarin dose not be changed for isolated, asymptomatic INRs of < or = 3.4; (2) to compare the dosing strategies of an ACS and primary care providers (PCPs); and (3) to quantify the relationship between reduction of the warfarin dose and the subsequent fall in the INR. DESIGN AND SETTING: Randomized controlled study of health maintenance organization outpatients who were receiving warfarin. PATIENTS: We identified 231 patients with a target INR of 2.5 and an isolated, asymptomatic INR between 3.2 and 3.4. Our ACS monitored 103 of the patients; PCPs monitored the remaining 128 patients. MEASUREMENTS: From all 231 patients, we obtained INRs and warfarin dosing history. From the 103 ACS enrollees, we also recorded adverse events. RESULTS: One ACS patient had epistaxis in the 30 days after the elevated INR. Twenty-three percent of ACS enrollees and 47% of PCP patients reduced their warfarin dose (p < 0.001). The median follow-up INRs were similar in both cohorts: 2.7 in the ACS enrollees and 2.6 in the PCP patients. However, in a subgroup analysis of 190 patients who presented with an INR of 3.2 or 3.3, ACS enrollees were more likely to have a follow-up INR in the range of 2 to 3 (p = 0.03). The median follow-up INR was 2.7 in 148 patients who maintained their warfarin dose, 2.5 in 77 patients who decreased their dose by 1 to 20%, and 1.7 in 6 patients who decreased their dose by 21 to 43% (p < 0.001). CONCLUSIONS: These findings support maintaining the same warfarin dose in asymptomatic patients with an INR of < or = 3.3, and reducing the dose for patients who have a greater INR or an increased risk of hemorrhage. Warfarin dose reductions > 20% should be avoided for mildly elevated INRs. PMID- 12576373 TI - Respiratory failure and sepsis are the major causes of ICU admissions and mortality in survivors of lung transplants. AB - STUDY OBJECTIVE: To identify outcome predictors and prognostic factors in long term survivors (> 30 days post-transplant) of single-lung and double-lung transplants on readmission to the medical ICU (MICU). DESIGN: Retrospective study. SETTING: MICU of the Cleveland Clinic Foundation, a lung transplantation center. PATIENTS: As of August 2000, 210 lung transplantations have been performed at our institution. The records of 33 lung transplant recipients who required readmission to the MICU after the initial 30-day post-transplant period over a 4-year period from August 16, 1996 to August 15, 2000 were reviewed. RESULTS: Thirty-three patients had a total of 46 MICU readmissions. Twenty-seven MICU admissions (59%) were due to respiratory deterioration with mechanical ventilation (10 deaths), and 16 MICU admissions (35%) were due to the systemic inflammatory response syndrome (SIRS; 8 deaths). The MICU mortality rate was 37% per admission in our group of patients. A preadmission diagnosis of bronchiolitis obliterans syndrome was observed in 7 of 14 nonsurvivors (50%) and in 5 of 25 patients (20%) surviving to hospital discharge. CONCLUSIONS: Respiratory failure and SIRS are the predominant causes of MICU readmissions and are frequent causes of death. APACHE (acute physiology and chronic health evaluation) III scores, nonpulmonary organ system dysfunction, initial serum albumin level, and duration of mechanical ventilation are important prognostic factors. PMID- 12576374 TI - Heroin insufflation as a trigger for patients with life-threatening asthma. AB - STUDY OBJECTIVES: To determine the prevalence of self-reported, heroin-associated asthma symptoms among inner-city patients treated for life-threatening asthma, and to compare the rates of drug use between ICU patients with asthma and ICU control patients with diabetic ketoacidosis (DKA). DESIGNS: Study 1 was a sequential case series of patients requiring ICU admission for asthma (January to June 1999). Study 2 was a retrospective, case-control study of drug use among asthma patients and control subjects with DKA requiring ICU care (1997 to 1998). SETTING: Inner-city, public hospital ICU. PATIENTS: Twenty-three patients (26 ICU admissions) with asthma (age range, 16 to 50 years) admitted to the ICU from January to June 1999, and 84 patients (104 ICU admissions) with asthma and 42 patients with DKA (age range, 15 to 50 years) admitted to the ICU during 1997 to 1998. Outcomes studied: Self-reported, heroin-associated exacerbations, history of heroin or cocaine use, and urine drug screen (UDS) results. MEASUREMENTS AND RESULTS: In the sequential ICU admissions, 13 of 23 patients (56%) described asthma exacerbations associated with heroin insufflation. In the case-control study, asthmatics were significantly more likely to report heroin use (41.3% vs 12.5%; p = 0.006) and had a significantly higher prevalence of UDS results positive for opiates (60% vs 7%; p = 0.001) compared to subjects with DKA. The rates of cocaine use by history and UDS results did not differ significantly between the two groups. CONCLUSIONS: At least since 1997, heroin insufflation is a common asthma trigger in this inner-city ICU and should be considered in the care of patients with life-threatening asthma. PMID- 12576375 TI - Microscopic examination of intracellular organisms in protected bronchoalveolar mini-lavage fluid for the diagnosis of ventilator-associated pneumonia. AB - STUDY OBJECTIVES: To assess the cutoff percentage of cells containing intracellular organisms (ICOs) in the microscopic examination of mini-lavage fluid for the diagnosis of ventilator-associated pneumonia (VAP), and to study the accuracy of this diagnostic procedure on patients who have received previous antibiotic therapy (AT). DESIGN: Prospective clinical investigation. SETTING: The medico-surgical ICU of a university hospital. PATIENTS: Eighty-two patients who fulfilled the clinical criteria of first episode of VAP. INTERVENTIONS: Lower airway secretion samples were obtained by a nonbronchoscopic protected bronchoalveolar mini-lavage (mini-PBAL). MEASUREMENTS AND RESULTS: A total of 82 mini-PBALs were performed. The fluid obtained was divided into two samples. The first sample underwent direct microscopic examination using centrifugal cytology and Giemsa stain in order to determine the percentage of cells containing ICOs. The second sample was processed for Gram stain and quantitative cultures. VAP was the final diagnosis in 65 patients based on the mini-PBAL results obtained in the quantitative cultures. At least one bacterial species grew in a significant concentration (> or = 10(3) cfu/mL). The mini-PBAL was performed on 42 patients during AT (> or = 24 h of effective AT at the time of diagnostic procedure) and on another 40 patients with no AT (No AT). The results of the quantitative cultures were compared with the percentage of cells containing ICOs using a receiver operator characteristic (ROC) curve. The cutoff point of > or = 2% of cells containing ICOs had the highest sensitivity (80%) and specificity (82%) in the studied population (area under the ROC curve [AUC], 0.83; 95% confidence interval [CI], 0.70 to 0.90). In patients receiving AT, the sensitivity was 70% and specificity was 75% (AUC, 0.73; 95% CI, 0.58 to 0.90); and in No AT-group patients sensitivity was 88% and specificity was 100% (AUC, 0.92; 95% CI, 0.84 to 1.0). The comparative analysis of both ROC curves was statistically significant (p = 0.04). CONCLUSIONS: The cutoff point of > or = 2% of cells containing ICOs has the highest sensitivity and specificity in the microscopic examination of mini-lavage fluid for the diagnosis of VAP. However, sensitivity is too low to be clinically useful. The direct examination of mini-PBAL fluid is less accurate when previous AT has been administered. PMID- 12576376 TI - Neurokinin-1 and neurokinin-2 antagonism inhibits long-term acid fog-induced airway hyperresponsiveness. AB - BACKGROUND: We recently reported that airway hyperresponsiveness (AHR) induced by a 6-h exposure to sulfuric acid (H(2)SO(4)) was inhibited by either the neurokinin (NK)-1 receptor antagonist, FK888, or the NK-2 receptor antagonist, SR48968, when administered immediately before the exposure. The aims of this study were to determine whether these antagonists have any therapeutic efficiency against AHR after long-term H(2)SO(4) inhalation and to elucidate the mechanisms in ovalbumin sensitized guinea pigs. METHODS: Specific airway resistance (sRaw), AHR, and BAL fluid were analyzed after an 8-week exposure to H(2)SO(4) aerosol (82 mg/m(3), pH 1.7, 40 mOsm) or hypotonic saline solution (pH 5.9, 40 mOsm) as a control. The H(2)SO(4) group then received a 2-week treatment with FK888, SR48968, or vehicle. RESULTS: The AHR and the eosinophil count in BAL fluid were significantly increased in the H(2)SO(4) group compared to control animals, while sRaw was significantly elevated in both groups after the 8-week exposure. Treatment with both FK888 and SR48968 significantly reduced the AHR and tended to inhibit eosinophilia in BAL fluid, but sRaw did not change. The degree of AHR improvement with SR48968 was much larger than with FK888. CONCLUSION: Our results show that both NK-1 and NK-2 receptor antagonists inhibited long-term H(2)SO(4) induced AHR in sensitized guinea pigs, and the effect was much greater with an NK 2 antagonist. We suggest that NK-1 or NK-2 antagonism might partially inhibit the H(2)SO(4)-induced influx of eosinophils into the lung. PMID- 12576377 TI - Neonatal exposure to 65% oxygen durably impairs lung architecture and breathing pattern in adult mice. AB - STUDY OBJECTIVE: To test the hypothesis that exposure to hyperoxia during the postnatal period of rapid alveolar multiplication by septation would cause permanent impairments, even with moderate levels of hyperoxia. DESIGN: We exposed mouse pups to 65% O(2) (hyperoxic mice) or normoxia (normoxic mice) during their first postnatal month, and we analyzed lung histology, pulmonary mechanics, blood gas, and breathing pattern during normoxia or in response to chemical stimuli in adulthood, when they reached 7 to 8 months of postnatal age. RESULTS: Hyperoxic mice had fewer and larger alveoli than normoxic mice (number of alveoli per unit surface area of parenchyma, 266 +/- 62/mm(2) vs 578 +/- 77/mm(2), p < 0.0001) [mean +/- SD], the cause being impaired alveolarization (radial alveolar count, 5.8 +/- 0.2 in hyperoxic mice vs 10.5 +/- 0.5 in normoxic mice, p < 0.0001). Respiratory system compliance was higher in hyperoxic mice (0.098 +/- 0.006 mL/cm H(2)O) than in normoxic mice (0.064 +/- 0.006 mL/cm H(2)O, p < 0.016). Baseline tidal volume (VT) and breath duration (TTOT]) measured noninvasively by whole body plethysmography were larger in hyperoxic mice than in normoxic mice (VT, + 15%, p < 0.01; TTOT, + 12%, p < 0.01). Despite these impairments, blood gas, baseline minute ventilation E, and E responses to hypoxia and hypercapnia were normal in hyperoxic mice, compared with normoxic mice. CONCLUSION: Hyperoxic exposure during lung septation in mice may cause irreversible lung injury and breathing pattern abnormalities in adulthood at O(2) concentrations lower than previously thought. However, ventilatory function and body growth were preserved, and ventilatory function showed no major abnormalities, at least at rest, despite early oxygen-induced injuries. PMID- 12576378 TI - Atrial fibrillation is associated with accumulation of aging-related common type mitochondrial DNA deletion mutation in human atrial tissue. AB - STUDY OBJECTIVE: Accumulation of somatic mutations of mitochondrial DNA (mtDNA) contributes to the aging process and progressive organ dysfunction. We investigated the mitochondrial DNA with 4977-base-pair mtDNA deletion mutation (mtDNA(4977)) in human atrial tissue and correlated the amount of mtDNA(4977) to clinical atrial fibrillation (AF). METHODS AND RESULTS: Atrial tissue from the right atrial appendage was obtained in 88 patients during open-heart surgery (22 children/adolescents and 66 adults). The amount of mtDNA(4977) was measured using a nested polymerase chain reaction protocol and normalized to wild-type mtDNA. We found that the mtDNA(4977) was absent in all 22 pediatric/adolescent patients. In the adult group, the relative amount of mtDNA(4977) was significantly higher in patients with AF than in patients without AF (0.55 +/- 0.26 vs 0.35 +/- 0.29, p < 0.007) [mean +/- SD]. The amount of mtDNA(4977) was also positively associated with age (r = 0.29, p < 0.01). Left and right atrial pressures, left atrial dimension, hypertension, and cardiac diagnosis did not influence the amount of mtDNA(4977) significantly. Further multivariate analysis showed that both aging and AF contributed independently to the accumulation of mtDNA(4977). CONCLUSION: AF is associated with an increase of mtDNA(4977). This change is similar to the aging process of atrial tissue and might contribute to atrial dysfunction in AF. PMID- 12576379 TI - Detection of loss of heterozygosity by high-resolution fluorescent system in non small cell lung cancer: association of loss of heterozygosity with smoking and tumor progression. AB - BACKGROUND: We recently developed a novel system for detecting microsatellite alteration, which is an important process in carcinogenesis. In patients with non small cell lung cancer (NSCLC), loss of heterozygosity (LOH) is frequently observed and causes functional disorders of tumor suppressor genes. PATIENTS AND METHODS: In a consecutive series of 51 patients with NSCLC who had undergone a surgical resection, microsatellite instability (MSI) and LOH in tumors were analyzed by polymerase chain reaction using five fluorescence-labeled dinucleotide markers (D2S123, D5S107, D10S197, D11SS904, and D13S175) and an autosequencer. RESULTS: MSI was detected in only one patient (2.0%) with only one marker. LOH was detected in at least one chromosomal region that was tested in 39 patients (76%). The mean (+/- SD) number of LOHs detected by each marker was 1.74 +/- 1.40, with 1 LOH detected in 10 patients, 2 LOHs detected in 15 patients, 10 LOHs detected in 3 patients, 1 LOH detected in 4 patients, and 3 LOHs detected in 5 patients. The number of LOHs detected in each patient was significantly associated with the pack-year index (rho = 0.501; p = 0.0004), although there was no relationship with having a history of multiple cancers and familial cancer. Patients with stage IA disease showed a significantly lower number of LOHs than did patients with other stages of disease (1.15 vs 2.38, respectively; p = 0.0013). CONCLUSION: LOH is very common in patients with NSCLC, and the number of LOHs increases with increases in smoking, suggesting the presence of an important event in lung carcinogenesis. PMID- 12576380 TI - Surgical treatment of malignant pleural mesothelioma: a review. AB - Despite many years of clinical research, there is still no effective therapy for malignant pleural mesothelioma (MPM). Untreated, the prognosis is poor, with a median survival of < 1 year. Single-agent or combination chemotherapy as well as radiotherapy have not shown persistent improvements in response or survival. In general, MPM is a disease confined to the pleural cavity for a long time before metastasizing. Therefore, focus on local treatment seems rational. Surgical resection has been considered the mainstay of treatment by some. However, surgery alone results in high recurrence rates, and the survival benefit remains questionable. In recent years, the emphasis has been on surgery combined with adjuvant therapies. In this article, the present state of surgical management of MPM will be reviewed. PMID- 12576381 TI - Portopulmonary hypertension: a tale of two circulations. AB - Pulmonary involvement is common in patients with portal hypertension and can manifest in diverse manners. Changes in pulmonary arterial resistance, manifesting either as the hepatopulmonary syndrome or portopulmonary hypertension (PPHTN), have been increasingly recognized in these patients in recent years. This review summarizes the clinicopathologic features, diagnostic criteria, as well as the latest concepts in the pathogenesis and management of PPHTN, which is defined as an elevated pulmonary artery pressure in the setting of an increased pulmonary vascular resistance and a normal wedge pressure in a patient with portal hypertension. PMID- 12576382 TI - Adult toxicology in critical care: part I: general approach to the intoxicated patient. AB - Intensivists are confronted with poisoned patients on a routine basis, with clinical scenarios ranging from known drug overdose or toxic exposure, illicit drug use, suicide attempt, or accidental exposure. In addition, drug toxicity can also manifest in hospitalized patients from inappropriate dosing and drug interactions. In this review article, we describe the epidemiology of poisoning in the United States, review physical examination findings and laboratory data that may aid the intensivist in recognizing a toxidrome (symptom complex of specific poisoning) or specific poisoning, and describe a rational and systematic approach to the poisoned patient. It is important to recognize that there is a paucity of evidence-based information on the management of poisoned patient. However, the most current recommendations by the American Academy of Clinical Toxicology and European Association of Poisons Centers and Clinical Toxicologists will be reviewed. Specific poisonings will be reviewed in the second section of these review articles. PMID- 12576383 TI - Top ten list in asthma. PMID- 12576384 TI - Accelerated decline of lung function in COPD patients with chronic hepatitis C virus infection: a preliminary study based on small numbers of patients. AB - STUDY OBJECTIVES: It has been suggested that chronic viral infection may increase the risk for development of COPD. This prospective study was designed to determine that chronic hepatitis C virus (HCV) infection is associated with accelerated decline of lung function in patients with COPD, and that antiviral therapy against HCV is effective for such patients. DESIGN: Prospective 5-year follow-up study. SETTING: University hospital. PATIENTS: Fifty-nine patients with COPD (group A, 15 HCV-negative ex-smokers; group B, 14 HCV-negative current smokers; group C, 14 HCV-positive ex-smokers; group D, 16 HCV-positive current smokers). INTERVENTIONS: After a 5-year follow-up period, 21 HCV-positive patients received interferon (IFN)-alpha therapy. MEASUREMENTS AND RESULTS: The rate of annual decline in FEV(1) and diffusing capacity of the lung for carbon monoxide (DLCO) during the 5-year follow-up period were significantly higher in group B (DeltaFEV(1), 59.7 mL/yr [SD, 17.5], p = 0.0008; DeltaDLCO, 3.50%/yr [SD, 0.44], p < 0.0001) and group C (DeltaFEV(1), 54.0 mL/yr [SD, 15.3], p = 0.0128; DeltaDLCO, 3.36%/yr [SD, 0.28], p < 0.0001) than in group A (DeltaFEV(1), 33.5 mL/yr [SD, 7.7]; DeltaDLCO, 2.66%/yr [SD, 0.34]). Moreover, these parameters in group D (DeltaFEV(1), 79.5 mL/yr [SD, 20.6]; DLCO, 4.5%/yr [SD, 0.40]) were also significantly higher than those in group B and group C. We evaluated the DeltaFEV(1) after IFN therapy during the 3-year follow-up period in the 8 IFN responders and 13 IFN nonresponders. DeltaFEV(1) in the IFN nonresponders did not significantly change during the 3-year follow-up period (before, 65.5 mL/yr [SD, 23.5]; after, 66.1 mL/yr [SD, 24.0]). However, DeltaFEV(1) in the IFN responders significantly decreased (before, 68.4 mL/yr [SD, 26.2]; after, 57.3 mL/yr [SD, 23.6], p = 0.0116). CONCLUSIONS: Our findings suggest that chronic HCV infection might accelerate decline in lung function in patients who already have COPD. PMID- 12576385 TI - Interferon therapy induces the improvement of lung function by inhaled corticosteroid therapy in asthmatic patients with chronic hepatitis C virus infection: a preliminary study. AB - STUDY OBJECTIVES: Several reports have suggested that subsets of asthmatic patients with chronic viral infection fail to respond to corticosteroid therapy. Therefore, this study was designed to determine that asthmatic patients with chronic hepatitis C virus (HCV) infection fail to improve lung function by inhaled corticosteroid therapy, and that interferon (IFN) therapy against HCV is effective for such patients. DESIGN: Prospective observational study. SETTING: University hospital. PATIENTS: Forty asthmatic patients with chronic HCV infection. INTERVENTIONS: After a 4-week run-in period, all asthmatic patients received therapy with inhaled beclomethasone dipropionate (BDP), 400 micro g twice daily for 6 weeks. After the first study, all asthmatic patients continued to receive inhaled BDP, and 30 HCV-positive asthmatic patients received IFN-alpha therapy for 6 months. MEASUREMENTS AND RESULTS: Prebronchodilator and postbronchodilator FEV(1) values were examined after a 4-week run-in period, after 6 weeks of BDP therapy, and at 1 year from the end of IFN therapy. After a 4-week run-in period as well as after 6 weeks of BDP therapy, there were no significant differences in either prebronchodilator or postbronchodilator FEV(1) values among the three groups. However, 1 year after the end of IFN therapy, the mean prebronchodilator and postbronchodilator FEV(1) values were significantly higher in the IFN responder group (n = 11) [prebronchodilator FEV(1), 1.93 L (SD, 0.13 L); postbronchodilator FEV(1), 2.28 L (SD, 0.15 L)] than in the IFN nontreatment group (n = 10) [prebronchodilator FEV(1), 1.78 L (SD, 0.10 L); p = 0.01; postbronchodilator FEV(1), 2.07 L (0.13 L); p = 0.005] or the IFN nonresponder groups (n = 19) [prebronchodilator FEV(1), 1.79 L (SD, 0.15 L); p = 0.006; postbronchodilator FEV(1), 2.07 L (SD, 0.18 L); p = 0.002]. Moreover, prebronchodilator and postbronchodilator FEV(1) values were significantly higher only in the IFN responder group at 1 year after the end of IFN therapy than after the 4-week run-in period (prebronchodilator FEV(1), p = 0.028; postbronchodilator FEV(1); p = 0.002) or after 6 weeks of BDP therapy (p = 0.016 and p = 0.004, respectively). CONCLUSIONS: Our findings suggest that chronic HCV infection in asthmatic patients is associated with impaired responses to inhaled BDP therapy and that intervention with IFN reverses such responses only in the IFN responder group. PMID- 12576386 TI - Ultrasound-guided transbronchial needle aspiration: an experience in 242 patients. AB - STUDY OBJECTIVES: Conventional transbronchial needle aspiration (TBNA) is a valuable procedure but remains underutilized. Recently, imaging guidance such as CT fluoroscopy has created considerable interest. As CT fluoroscopy is cumbersome and exposes patients and staff to radiation, we have examined the feasibility of endobronchial ultrasound (EBUS) in providing imaging support for TBNA. DESIGN: Prospective consecutive patient enrollment. SETTING: University-related tertiary referral centers for pulmonary diseases. RESULTS: From January 1999 to January 2000, 242 consecutive patients were entered into this prospective study (82 women and 160 men; mean age, 60.0 years). Indications for TBNA were diagnosis of enlarged lymph nodes and cancer staging. The average lymph node size was 1.7 cm (SD, 0.47; range, 0.8 to 4.3 cm). Target lymph nodes were visualized with EBUS, followed by TBNA in standard fashion. All targets could be visualized with EBUS. In 207 patients, the lymph nodes were successfully sampled (86%). This was independent of lymph node size and location. A firm diagnosis or cancer stage could be obtained in 172 patients (72%). There were no complications associated with the use of EBUS. CONCLUSION: EBUS is simply performed and if used for TBNA guidance affords an excellent yield independent of lymph node location. Randomized trials comparing standard TBNA and imaging-guided TBNA by CT fluoroscopy and EBUS are indicated. PMID- 12576387 TI - Fever, cough, and bilateral lung infiltrates. Achalasia associated with aspiration pneumonia. PMID- 12576388 TI - A 74-year-old woman with desaturation following surgery. Co-oximetry is the first step in making the diagnosis of dyshemoglobinemia. PMID- 12576389 TI - Hemoptysis, history of oral ulcers, and abnormal chest radiograph in a 64-year old man. PMID- 12576390 TI - Thoracoscopic lobectomy for lung cancer with a largely fused fissure. AB - BACKGROUND: While isolating the pulmonary arterial branches within the fissure is a crucial step in lobectomy, a largely fused fissure usually hinders its achievement, making lobectomy with video-assisted thoracoscopic surgery (VATS) difficult to achieve. For VATS lobectomy in lung cancer patients with a largely fused fissure, we have conducted an unusual approach for each lobe, and the surgical results were compared between patients with and without a fused fissure. METHODS: Since1999, we have conducted VATS lobectomies in 77 patients. Of these, 10 had largely fused fissures that needed an unusual surgical approach for dividing the pulmonary arterial branches. The other 67 patients had separated fissures that allowed the isolation and division of the arterial branches within it. While the surgical approach used for the patients with largely fused fissures differed in each lobe, most often the lobar bronchus was divided before pulmonary arterial branches within the fissure were divided, with the fused fissure being divided last. RESULTS: There were no significant differences in age, lobectomy site, or tumor stage between the patients with fused fissures and those with separated fissures. The surgical data showed no significant differences between the two groups in operating time, blood loss, duration of chest tube drainage, and hospital stay after surgery. However, the patients with fused fissures required more staples to close the incision than did those with a separated fissure (mean number of staples, 7.7 vs 5.7; p < 0.001). There was no postoperative mortality or morbidity, including prolonged air leakage, in the patients with fused fissures. CONCLUSION: Although the performance of VATS lobectomy for patients with largely fused fissures is more costly, it is feasible and safe. A largely fused fissure is not a limiting factor for the performance of VATS lobectomy. PMID- 12576391 TI - Chylothorax in lymphangioleiomyomatosis. AB - STUDY OBJECTIVE: Pulmonary lymphangioleiomyomatosis (LAM) is a rare, serious disorder characterized by proliferation of abnormal smooth-muscle cells and affects almost exclusively women of childbearing age. Optimal management of chylothorax, a well-recognized complication of LAM, in these patients has not been defined. This study was performed to characterize the clinical course and identify appropriate management options for chylothorax occurring in patients with LAM. DESIGN: Identification and retrospective review of available medical records on patients with LAM and chylothorax. SETTING: Tertiary-referral medical center. PATIENTS: All patients with LAM seen at Mayo Clinic, Rochester, MN, from January 1, 1976, to December 31, 2000. INTERVENTION: None. MEASUREMENT AND RESULTS: Eight of 79 patients (10.1%) with LAM had chylothorax. All were women aged 33 to 51 years, and four patients had underlying tuberous sclerosis complex. These eight women represented 3.5% of the 229 patients with chylothorax seen over this 25-year period at Mayo Clinic Rochester. Six patients had unilateral pleural effusion and two patients had bilateral effusions at initial presentation. The size of the chylothorax varied and was not necessarily progressive. Management of chylothorax ranged from thoracentesis only to thoracotomy with thoracic duct ligation and parietal pleurectomy. When needed, pleurodesis by instillation of sclerosing agents or parietal pleurectomy appeared to be effective in controlling chylothorax. CONCLUSIONS: Chylothorax occurring in patients with LAM has a variable clinical course. Although pleurodesis with or without thoracic duct ligation appears to be effective in controlling intractable chylothorax, less invasive treatments such as thoracentesis or observation may suffice in some cases. Management of chylothorax in patients with LAM should be individualized depending on the size and clinical effects of the chylous pleural effusion, as well as comorbid factors and local expertise. PMID- 12576392 TI - Paraneoplastic bilateral proptosis in a case of non-small cell lung cancer. AB - We report the case of a 51-year-old woman with a large cell bronchial carcinoma with neuroendocrine differentiation (T3N0M0, stage IIB) and bilateral exophthalmos. A CT of the orbitae was compatible with Graves ophthalmopathy, but neither abnormalities in thyroid function nor thyroid antibodies could be found. The proptosis normalized 10 days after tumor resection. We conclude that the most likely cause of the reversible proptosis in this patient is paraneoplastic, a condition not previously described in the English-language literature. PMID- 12576393 TI - Pulmonary eosinophilia following lung transplantation for sarcoidosis in two patients. AB - Pulmonary eosinophilia is an uncommon problem in lung transplant recipients. We report the unique occurrence of two cases of pulmonary eosinophilia in pulmonary allografts for sarcoidosis. Both patients rapidly acquired bronchiolitis obliterans syndrome (BOS) after resolution of pulmonary eosinophilia. It is known that peripheral eosinophilia is a marker for pulmonary allograft rejection, but its potential in the pathogenesis of BOS is unclear. PMID- 12576394 TI - Sleep-disordered breathing associated with long-term opioid therapy. AB - Three patients are described who illustrate distinctive patterns of sleep disordered breathing that we have observed in patients who are receiving long term, sustained-release opioid medications. Polysomnography shows respiratory disturbances occur predominantly during non-rapid eye movement (NREM) sleep and are characterized by ataxic breathing, central apneas, sustained hypoxemia, and unusually prolonged obstructive "hypopneas" secondary to delayed arousal responses. In contrast to what is usually observed in subjects with obstructive sleep apnea (OSA), oxygen desaturation is more severe and respiratory disturbances are longer during NREM sleep compared to rapid eye movement sleep. Further studies are needed regarding the effects of opioids on respiration during sleep as well as the importance of interaction with other medications and associated risk factors for OSA. PMID- 12576395 TI - Leptospirosis presenting as diffuse alveolar hemorrhage: case report and literature review. AB - The literature on diffuse alveolar hemorrhage heavily emphasizes the causal role of vasculitides. We present a patient with diffuse alveolar hemorrhage caused by leptospirosis. Although the pathology in leptospirosis occurs secondary to a vasculitic process, this disease is not listed as a cause of diffuse alveolar hemorrhage in the review literature. In the right clinical scenario, the disease should be considered in a patient presenting with diffuse alveolar hemorrhage. PMID- 12576396 TI - Tracheobronchial stenosis from acid aspiration presenting as asthma. AB - We report a case of a 42-year-old man who fell in a vat of hydrochloric acid, resulting in ingestion and aspiration of acid. Initially, he suffered from a chemical pneumonitis and GI burns. He was released from the hospital without complications, only to return with signs and symptoms consistent with asthma. Evaluation revealed multiple areas of large airway stenosis, resulting from the chemical burns. The stenoses were treated with multiple stents. PMID- 12576397 TI - Pulmonary toxicity in patients receiving low-dose amiodarone. AB - RATIONALE: Although there have been reports of pulmonary toxicity with low-dose amiodarone, it is generally believed that low-dose therapy is safe. METHODS: The clinical data for eight patients identified from a retrospective chart review are presented. RESULTS: All of the patients were receiving amiodarone, 200 mg/d, for an average of 2 years. The average age was 77 years (range, 65 to 89 years). Seven of the eight patients were male. Seven of the eight patients presented with dyspnea on exertion, and three of the eight patients presented with cough. All of the patients had a clinical diagnosis of amiodarone-induced pulmonary toxicity. Open-lung biopsies were obtained on two patients that were consistent with amiodarone-induced pulmonary toxicity. None of the patients were in congestive heart failure. Treatment involved cessation of amiodarone. In addition, three patients received corticosteroids. Five of the patients improved symptomatically with this conservative approach, and four patients improved radiographically. One patient died with progressive respiratory insufficiency (presumably from amiodarone pulmonary toxicity). One patient was unavailable for follow-up. CONCLUSION: Amiodarone-induced pulmonary toxicity can occur at a daily dose of 200 mg. Clinicians must remain alert to this possibility even with this low-dose therapy. PMID- 12576398 TI - Proteinuria in COPD patients with and without respiratory failure. PMID- 12576399 TI - Lung cancer screening debate. PMID- 12576400 TI - How to design a negative study. PMID- 12576401 TI - Endobronchial actinomycosis and foreign body. PMID- 12576402 TI - Identifying sleep apnea. PMID- 12576405 TI - Influence of sleep state on frequency of swallowing, apnea, and arousal in human infants. AB - Apnea and arousal are modulated with sleep stage, and swallowing may interfere with respiratory rhythm in infants. We hypothesized that swallowing itself would display interaction with sleep state. Concurrent polysomnography and measurement of swallowing allowed time-matched analysis of 3,092 swallows, 482 apneas, and 771 arousals in 17 infants aged 1-34 wk. The mean rates of swallowing, apnea, and arousal were significantly different, being 23.3 +/- 8.5, 9.4 +/- 8.8, and 15.5 +/- 10.6 h(-1), respectively (P < 0.001 ANOVA). Swallows occurred before 25.2 +/- 7.9% and during 74.8 +/- 6.3% of apneas and before 39.8 +/- 6.0% and during 60.2 +/- 6.0% of arousals. The frequencies of apneas and arousals were both strongly influenced by sleep state (active sleep > indeterminate > quiet sleep, P < 0.001), whether or not the events coincided with swallowing, but swallowing rate showed minimal independent interaction with sleep state. Interactions between swallowing and sleep state were predominantly influenced by the coincidence of swallowing with apnea or arousal. PMID- 12576406 TI - Effect of stimulus cycle time on acute respiratory responses to intermittent hypercapnic hypoxia in unsedated piglets. AB - To determine whether stimulus frequency affects physiological compensation to an intermittent respiratory stimulus, we studied piglets (n = 43) aged 14.8 +/- 2.4 days. A 24-min total hypercapnic hypoxia (HH) (10% O(2)-6% CO(2)-balance N(2) = HH) was delivered in 24-, 8-, 4-, or 2-min cycles alternating with air. Controls (n = 10) breathed air continuously. Minute ventilation and temperature were not different between the 2-min and 24-min groups, with neither different from controls during recovery. Piglets exposed to 8-min cycles had ventilatory stimulation, whereas those exposed to 4-min cycles had significant depression of ventilation. Despite this, piglets in these intermediate intermittent HH (IHH) groups (8- and 4-min cycles) showed more severe acidosis and attenuated temperature changes (P < 0.001 and P < 0.01 for pH and temperature vs. 24 min, respectively). Cycle time affected the ability of young piglets to tolerate IHH. More severe respiratory acidosis developed when IHH was delivered in intermediate (4 min or 8 min) cycles compared with the same total dose as a single episode or in short (2 min) cycles. PMID- 12576407 TI - Speech movements do not scale by orofacial structure size. AB - The potential role of a size-scaling principle in orofacial movements for speech was examined by using between-group (adults vs. 5-yr-old children) as well as within-group correlational analyses. Movements of the lower lip and jaw were recorded during speech production, and anthropometric measures of orofacial structures were made. Adult women produced speech movements of equal amplitude and velocity to those of adult men. The children produced speech movement amplitudes equal to those of adults, but they had significantly lower peak velocities of orofacial movement. Thus we found no evidence supporting a size scaling principle for orofacial speech movements. Young children have a relatively large-amplitude, low-velocity movement strategy for speech production compared with young adults. This strategy may reflect the need for more time to plan speech movement sequences and an increased reliance on sensory feedback as young children develop speech motor control processes. PMID- 12576408 TI - Hypercapnic cerebral vascular reactivity is decreased, in humans, during sleep compared with wakefulness. AB - During wakefulness, increases in the partial pressure of arterial CO(2) result in marked rises in cortical blood flow. However, during stage III-IV, non-rapid eye movement (NREM) sleep, and despite a relative state of hypercapnia, cortical blood flow is reduced compared with wakefulness. In the present study, we tested the hypothesis that, in normal subjects, hypercapnic cerebral vascular reactivity is decreased during stage III-IV NREM sleep compared with wakefulness. A 2-MHz pulsed Doppler ultrasound system was used to measure the left middle cerebral artery velocity (MCAV; cm/s) in 12 healthy individuals while awake and during stage III-IV NREM sleep. The end-tidal Pco(2) (Pet(CO(2))) was elevated during the awake and sleep states by regulating the inspired CO(2) load. The cerebral vascular reactivity to CO(2) was calculated from the relationship between Pet(CO(2)) and MCAV by using linear regression. From wakefulness to sleep, the Pet(CO(2)) increased by 3.4 Torr (P < 0.001) and the MCAV fell by 11.7% (P < 0.001). A marked decrease in cerebral vascular reactivity was noted in all subjects, with an average fall of 70.1% (P = 0.001). This decrease in hypercapnic cerebral vascular reactivity may, at least in part, explain the stage III-IV NREM sleep-related reduction in cortical blood flow. PMID- 12576409 TI - Lesions of rat skeletal muscle after local block of acetylcholinesterase and neuromuscular stimulation. AB - In skeletal muscle, a local increase of acetylcholine (ACh) in a few end plates has been hypothesized to cause the formation of contraction knots that can be found in myofascial trigger points. To test this hypothesis in rats, small amounts of an acetylcholinesterase inhibitor [diisopropylfluorophosphate (DFP)] were injected into the proximal half of the gastrocnemius muscle, and the muscle nerve was electrically stimulated for 30-60 min for induction of muscle twitches. The distal half of the muscle, which performed the same contractions, served as a control to assess the effects of the twitches without DFP. Sections of the muscle were evaluated for morphological changes in relation to the location of blocked end plates. Compared with the distal half of the muscle, the DFP-injected proximal half exhibited significantly higher numbers of abnormally contracted fibers (local contractures), torn fibers, and longitudinal stripes. DFP-injected animals in which the muscle nerve was not stimulated and that were allowed to survive for 24 h exhibited the same lesions but in smaller numbers. The data indicate that an increased concentration of ACh in a few end plates causes damage to muscle fibers. The results support the assumption that a dysfunctional end plate exhibiting increased release of ACh may be the starting point for regional abnormal contractions, which are thought to be essential for the formation of myofascial trigger points. PMID- 12576410 TI - Contractile properties of rat single muscle fibers and myosin and troponin isoform expression after hypergravity. AB - The effects of 19 days of hypergravity (HG) were investigated on the biochemical and physiological properties of the slow soleus muscle and its fast agonist, the plantaris. HG was induced by rotational centrifugation that led to a 2-G gravity level. The HG rats were characterized by a slower body growth than control, whereas the soleus muscle mass was increased by 15%. Using electrophoretic techniques, we showed that the distribution of myosin heavy chain and troponin T isoforms was not modified after HG in both soleus and plantaris. In contrast, the isoform expression pattern of two troponin subunits, troponin I and troponin C, was changed in a slow-to-fast manner only in the soleus. From tension-pCa relationships, changes in Ca(2+) activation threshold by 0.18 pCa unit indicated a decrease in Ca(2+) sensitivity and an increase in the slope of the curve, attesting to a higher cooperativity along the thin filament after HG. Comparison of our HG data with previous results in microgravity conditions indicated that muscle characteristics, except muscle mass, did not evolve linearly from 0 to 2 G. PMID- 12576411 TI - Gender differences in skeletal muscle fatigability are related to contraction type and EMG spectral compression. AB - The purposes of this study were 1) to evaluate gender differences in back extensor endurance capacity during isometric and isotonic muscular contractions, 2) to determine the relation between absolute load and endurance time, and 3) to compare men [n = 10, age 22.4 +/- 0.69 (SE) yr] and women (n = 10, age 21.7 +/- 1.07 yr) in terms of neuromuscular activation patterns and median frequency (MF) shifts in the electromyogram (EMG) power spectrum of the lumbar and hip extensor muscles during fatiguing submaximal isometric trunk extension exercise. Subjects performed isotonic and isometric trunk extension exercise to muscular failure at 50% of maximum voluntary contraction force. Women exhibited a longer endurance time than men during the isometric task (146.0 +/- 10.9 vs. 105.4 +/- 7.9 s), but there was no difference in endurance performance during the isotonic exercise (24.3 +/- 3.4 vs. 24.0 +/- 2.8 repetitions). Absolute load was significantly related to isometric endurance time in the pooled sample (R(2) = 0.34) but not when men and women were analyzed separately (R(2) = 0.05 and 0.04, respectively). EMG data showed no differences in neuromuscular activation patterns; however, gender differences in MF shifts were observed. Women demonstrated a similar fatigability in the biceps femoris and lumbar extensors, whereas in men, the fatigability was more pronounced in the lumbar musculature than in the biceps femoris. Additionally, the MF of the lumbar extensors demonstrated a greater association with endurance time in men than in women (R(2) = 0.45 vs. 0.19). These findings suggest that gender differences in muscle fatigue are influenced by muscle contraction type and frequency shifts in the EMG signal but not by alterations in the synergistic activation patterns. PMID- 12576412 TI - Attenuated responses to sympathoexcitation in individuals with Down syndrome. AB - This study evaluated blood pressure and heart rate responses to exercise and nonexercise tasks as indexes of autonomic function in subjects with and without Down syndrome (DS). Twenty-four subjects (12 with and 12 without DS) completed maximal treadmill exercise, isometric handgrip (30% of maximum), and cold pressor tests, with heart rate and blood pressure measurements. Maximal heart rate and heart rate and blood pressure responses to the isometric handgrip and cold pressor tests were reduced in subjects with DS (P < 0.05). Both early (first 30 s) and late (last 30 s) responses were reduced. Obesity did not appear to influence the results, as both obese and normal-weight subjects with DS exhibited similar responses, and controlling for body mass index did not alter the results between controls and subjects with DS. Individuals with DS, without congenital heart disease, exhibit reduced heart rate and blood pressure responses to isometric handgrip exercise and cold pressor testing, consistent with autonomic dysfunction. Autonomic dysfunction may partially explain chronotropic incompetence observed during maximal treadmill exercise in individuals with DS. PMID- 12576413 TI - Power spectral and Poincare plot characteristics in sinus node dysfunction. AB - A salient feature of the normal sinus node activity is its prominent beat-to-beat variability, which shows self-similarity on different time scales (fractal dynamics). However, in patients with sinus node dysfunction, short-term time sinus cycles show exaggerated variability, the characteristics of which have not been analyzed. Therefore, Poincare plots and power spectral analysis were applied to short-term variations of sinus cycles in 30 patients with and 30 patients without sinus node disease. Three patterns of behavior were observed in sick sinus patients: type 1, completely normal (n = 3); type 2, randomlike pattern in the Poincare plots with "white noise" power spectra (n = 9); and type 3, a transitional pattern, characterized by remnants of normal behavior mixed with scattered points (n = 18). In control subjects, only type 1 (n = 27) and type 3 (n = 3) patterns were observed, P < 0.0001. The power spectral changes in sinus node dysfunction are thus characterized by a loss of the inverse power law relationship, which both has implications for heart rate variability analysis and might offer a new diagnostic approach. PMID- 12576414 TI - Temporal association of nitric oxide levels and airflow in asthma after whole lung allergen challenge. AB - Exhaled nitric oxide (NO) levels are high in asthmatic subjects and increase with exacerbations. We hypothesized that higher levels of NO observed during asthma exacerbations are due to increased synthesis of NO. Exhaled NO and peak flows were measured in 11 asthmatic and 9 healthy control subjects before and after experimental asthmatic response induced by whole lung allergen challenge. Baseline peak flows of asthmatics were significantly lower than controls and decreased significantly immediately after challenge (P = 0.004). NO was measured by collecting exhaled breaths without breath hold (NO0) and after a 15-s breath hold (NO15). The rate of NO accumulation over time [parts/billion per second (ppb/s)] was calculated by DeltaNO/Deltat = (NO15 - NO0)/15, where Delta denotes a change and t is time. The NO accumulation rates in asthmatic and control subjects were similar at baseline; however, NO accumulation at 24 h increased threefold from baseline in asthmatic compared with control subjects (asthmatic subjects, 0.6 +/- 0.2 ppb/s; control subjects, 0.2 +/- 0.1 ppb/s; P = 0.01). Our study suggests that increased NO during an asthma exacerbation is due to increased synthesis, perhaps by increased expression of NO synthases. PMID- 12576415 TI - Prognostic factors in multiple myeloma: it's in the genes. PMID- 12576416 TI - Modulation of cellular signaling pathways: prospects for targeted therapy in hematological malignancies. AB - The high remission rates observed in patients with chronic myelogenous leukemia who receive Imatinib mesylate (Gleevec) indicate that targeted therapy for hematological malignancies is achievable. At the same time, progress in cellular biology over the past decade has resulted in a better understanding of the process of malignant transformation, a better classification of subtypes of each disease on the basis of molecular markers, and a better characterization of the molecular targets for drug development. These advances have already spawned the development of such effective agents as monoclonal antibodies and specific enzyme inhibitors. This review attempts to provide a practical introduction to the complex and growing field of targeted therapy in hematological malignancies. PMID- 12576417 TI - Matrix metalloproteinases, angiogenesis, and cancer: commentary re: A. C. Lockhart et al., Reduction of wound angiogenesis in patients treated with BMS 275291, a broad spectrum matrix metalloproteinase inhibitor. Clin. Cancer Res., 9: 00-00, 2003. PMID- 12576418 TI - A phase I/II trial of intratumoral endoscopic ultrasound injection of ONYX-015 with intravenous gemcitabine in unresectable pancreatic carcinoma. AB - PURPOSE: Localized pancreatic carcinoma is rarely resectable and is resistant to conventional therapies. ONYX-015 (dl1520) is an E1B-55kD gene-deleted replication selective adenovirus that preferentially replicates in and kills malignant cells. Endoscopic ultrasound (EUS) has the potential to conveniently and accurately deliver local therapy to the pancreas. Therefore, we undertook a trial of the feasibility, tolerability, and efficacy of EUS injection of ONYX-015 into unresectable pancreatic carcinomas. EXPERIMENTAL DESIGN: Twenty-one patients with locally advanced adenocarcinoma of the pancreas or with metastatic disease, but minimal or absent liver metastases, underwent eight sessions of ONYX-015 delivered by EUS injection into the primary pancreatic tumor over 8 weeks. The final four treatments were given in combination with gemcitabine (i.v., 1,000 mg/m(2)). Patients received 2 x 10(10) (n = 3) or 2 x 10(11) (n = 18) virus particles/treatment. RESULTS: After combination therapy, 2 patients had partial regressions of the injected tumor, 2 had minor responses, 6 had stable disease, and 11 had progressive disease or had to go off study because of treatment toxicity. No clinical pancreatitis occurred despite mild, transient elevations in lipase in a minority of patients. Two patients had sepsis before the institution of prophylactic oral antibiotics. Two patients had duodenal perforations from the rigid endoscope tip. No perforations occurred after the protocol was changed to transgastic injections only. CONCLUSIONS: This study indicates that ONYX-015 injection via EUS into pancreatic carcinomas by the transgastic route with prophylactic antibiotics is feasible and generally well tolerated either alone or in combination with gemcitabine. Transgastric EUS-guided injection is a new and practical method of delivering biological agents to pancreatic tumors. PMID- 12576419 TI - Flavopiridol-related proinflammatory syndrome is associated with induction of interleukin-6. AB - BACKGROUND: Flavopiridol is a flavonoid with antiproliferative effects mediated, in part, by inhibition of cyclin-dependent kinases. Clinical manifestations in a previous Phase I trial in patients with refractory malignancies treated with a 72 h flavopiridol infusion included a proinflammatory syndrome consisting of fever, fatigue, and "local" tumor pain with concomitant alterations in plasma acute phase reactant proteins. PURPOSE: The aim of this study was to determine whether the proinflammatory syndrome observed in this trial was associated with modulation of plasma cytokines. METHODS: Patients receiving flavopiridol (n = 76) had serial plasma samples drawn preinfusion and during the infusion for evaluation of interleukin (IL)-6, IL-10, IL-12, granulocyte macrophage colony stimulating factor, basic-fibroblast growth factor, transforming growth factor beta, and tumor necrosis factor-alpha levels by standard ELISA assays. The Wilcoxon signed rank test was used to test the significance of the difference between the baseline (time 0) plasma cytokine levels compared with the values of each subsequent data collection time points (8, 24, 48, and 72 h). RESULTS: There was a significant and sustained increase in plasma IL-6 levels at all time points when compared with baseline values. Paired values were used in the statistical analysis. Median plasma (interquartile range) values of IL-6 were elevated from 15.5 (9-52) pg/ml at baseline to 23 (4-48) pg/ml (P < 0.01) at 8 h; from 15 (2 48) pg/ml at baseline to 46 (21-105) pg/ml (P < 0.001) at 24 h; from 16 (9-52) pg/ml at baseline to 61 (32-170) pg/ml (P < 0.001) at 48 h; and from 15.5 (6-48) pg/ml to 68 (40-200) pg/ml (P < 0.001) at 72 h. Significance was maintained even when adjusted for multiple comparisons. The relative increase in IL-6 concentration was dose-dependent. Moreover, IL-6 elevation had a direct correlation with flavopiridol peak plasma concentration, flavopiridol area under the curve, and plasma C-Reactive protein levels. A significant decrease in plasma granulocyte macrophage colony-stimulating factor occurred at the 8-h sampling point: 50 pg/ml (interquartile range 10-205 pg/ml, P < 0.01) when compared with baseline plasma levels and 71 pg/ml (interquartile range 5-152 pg/ml, P < 0.01). No changes in the other pro or anti-inflammatory cytokines were observed. Immunohistochemistry studies in bone marrow aspirates from a prospective group of patients in this trial demonstrated approximately 4-fold induction of IL-6 (compared with baseline), mostly in non-T cells. CONCLUSION: Biochemical analysis of plasma in patients undergoing infusional flavopiridol found a significant dose dependent induction of IL-6. IL-6 elevation could be a marker for the process leading to the appearance of the proinflammatory syndrome observed in patients treated with infusional flavopiridol. The mechanism(s) underlying IL-6 induction and its significance are still unknown but may influence strategies to modulate flavopiridol's clinical effects. PMID- 12576420 TI - Immunoscintigraphic detection of the ED-B domain of fibronectin, a marker of angiogenesis, in patients with cancer. AB - PURPOSE: ED-B fibronectin is expressed only during angiogenic processes and in tissues undergoing growth and/or extensive remodeling. We demonstrated previously the possibility to target and selectively deliver therapeutic substances to tumor vasculature in experimental animal models using a human recombinant antibody fragment, L19, specific for the ED-B domain of fibronectin. Here we evaluate the possibility of targeting primary tumors and metastatic lesions in cancer patients through immunoscintigraphy using (123)I-labeled dimeric L19 [L19(scFv)(2)]. EXPERIMENTAL DESIGN: Twenty patients (34-79 years of age) with lung, colorectal, or brain cancer, whose tumors had been confirmed by imaging techniques and/or histologically, were admitted to the immunoscintigraphic investigation. RESULTS: The dimeric L19 antibody selectively localized in tumor lesions in aggressive types of lung cancer and colorectal cancer. Because ED-B fibronectin is expressed only during angiogenic processes and in tissues undergoing growth and/or extensive remodeling, L19(scFv)(2) is able to distinguish between quiescent and actively growing lesions. No side effects were observed. CONCLUSIONS: The ability of L19(scFv)(2) to target tumors in patients provides the foundations for new therapeutic applications, in which the L19 antibody is engineered to selectively deliver bioactive molecules to primary tumors as well as to metastases. PMID- 12576421 TI - Capecitabine induces rapid, sustained response in two patients with extensive oral verrucous carcinoma. AB - PURPOSE: Oral verrucous carcinoma (VC) has been traditionally treated with surgery or radiation with frequent recurrences and significant morbidity. We describe rapid and dramatic response to oral capecitabine in two patients with advanced refractory VC. EXPERIMENTAL DESIGN: VC is a rare tumor of the oral cavity. It does not metastasize but over time causes morbidity and mortality through local invasion. Radiation and surgery have been the main treatment modalities but are plagued by mutilating effects, recurrences, and possibly malignant degeneration in some cases. To date, no effective chemotherapy regimens have been described in the international literature. The clinical records of two elderly females with extensive oral VC are described. Both patients were prescribed one cycle of capecitabine, 1,000 mg bid for 14 days. Response was documented by photography in one patient. Immunohistochemical evaluation of three 5-fluorouracil metabolizing enzymes on pretreatment biopsies from both patients was also performed. A review of the literature with emphasis on treatment of oral VC is presented in view of our findings. RESULTS: Examination of the oral cavity before treatment revealed extensive involvement with an evenly spreading, exophytic, warty, whitish lesion in both patients. Microscopic examination of H&E stained slides from biopsies of these lesions confirmed the clinical suspicion of VC. Both patients underwent treatment with oral capecitabine for one cycle (2 weeks on/1 week off) at a reduced dose of 1000 mg, p.o., bid. Both had a dramatic response with near complete resolution of their lesions within 3 weeks of initiating therapy. A durable partial response was documented at 1 year in the first patient and 6 months in the second. Immunohistochemical evaluation of pretreatment biopsies from both patients revealed a high level of expression of thymidine phosphorylase, a key enzyme in the metabolism of capecitabine. CONCLUSIONS: Oral VC is a rare entity with a progressive course over years and limited options in terms of treatment. Preliminary observations in two elderly patients demonstrate that capecitabine, an oral fluoropyrimidine, is well tolerated and may induce rapid, clinically significant response. Although not curative, it may provide a cost-effective alternative for elderly patients with a significant improvement in their quality of life. PMID- 12576422 TI - Reduction of wound angiogenesis in patients treated with BMS-275291, a broad spectrum matrix metalloproteinase inhibitor. AB - PURPOSE: The purpose of this study was to evaluate the feasibility of incorporating a novel wound angiogenesis assay into a Phase I study of BMS 275291, a broad-spectrum matrix metalloproteinase inhibitor, and to determine whether the wound angiogenesis assay was able to detect the inhibition of angiogenesis in patients treated with BMS-275291. EXPERIMENTAL DESIGN: Before treatment began, a 4-mm skin biopsy was performed. The wound was imaged for 14 days. Treatment was started on day 0, and a separate 4-mm biopsy was performed 14 days later. The second wound was also imaged for 14 days. Wound angiogenesis was scored by two independent observers who were blinded to treatment status. RESULTS: The median times in days (95% confidence interval) to reach the target average vascular score (AVS) of 1.5 and 2.0 based on the data of Observer 1 were 3.7 (2.2-6.9) and 8.0 (5.0-10.0) pretreatment whereas on-treatment the values were 4.9 (3.7-8.0) and 9.3 (7.0-11.5), respectively. The delay in the median time to reach an AVS of 1.5 was 1.2 days or a 32% reduction when comparing pretreatment with on-treatment (P = 0.06). For the target AVS of 2.0 the delay in the median time pretreatment versus on-treatment was 1.3 days or a 16% reduction (P = 0.04). CONCLUSIONS: The wound angiogenesis assay used in this study was practical, well tolerated, and reproducible. Delays in wound angiogenesis because of BMS-275291 were detectable with this assay. This technique warrants additional investigation in clinical trials of other antiangiogenic agents. PMID- 12576423 TI - Tissue microarray analysis of signal transducers and activators of transcription 3 (Stat3) and phospho-Stat3 (Tyr705) in node-negative breast cancer shows nuclear localization is associated with a better prognosis. AB - PURPOSE: Although a high frequency of tumors contain constitutively activated signal transducers and activators of transcription 3 (Stat3), its relationship to breast cancer and patient survival has not been determined in a large retrospective study of node-negative tumors. To further elucidate the role of Stat3 in breast cancers, the expression patterns of Stat3 and Phospho-tyrosine residue 705 (Tyr705) Stat3 were correlated with survival outcome and clinicopathological parameters in a large cohort of node-negative breast cancer tumors. EXPERIMENTAL DESIGN: Immunohistochemical analysis of Stat3 and Phospho Stat3 was performed on a breast cancer tissue microarray of 346 node-negative breast cancer specimens. These results were correlated with overall survival and other clinicopathological data. RESULTS: Positive Stat3 cytoplasmic expression was seen in 69.2% of tumors, and positive Phospho-Stat3 (Tyr705) cytoplasmic expression was seen in 19.6% of tumors. Neither cytoplasmic expression showed significant association with survival or other clinical parameters. However, 23.1% of tumors had positive Stat3 nuclear expression, and those patients had a significantly improved short-term survival (P = 0.0332) at 5 years of follow-up. Upon analysis of positive Phospho-Stat3 (Tyr705) nuclear expression, seen in 43.5% of tumors, positive tumors had a significantly improved survival at both short-term 5-year survival (P = 0.0054) and long-term 20-year (P = 0.0376) survival analysis. Additionally, positive Phospho-Stat3 (Tyr705) nuclear expression is an independent prognostic marker of better overall survival node negative breast cancer by multivariate analyses that included the variables of nuclear grade, Ki-67, estrogen receptor staining, progesterone receptor staining, Her2 staining, age, and tumor size. CONCLUSIONS: These findings support a role for Stat3 and Phospho-Stat3 (Tyr705) overexpression in node-negative breast cancer and provide initial evidence that Phospho-Stat3 (Tyr705) may be a marker for improved overall survival independent of other prognostic markers. PMID- 12576424 TI - High and differential expression of HER-2/neu extracellular domain in bilateral ductal fluids from women with unilateral invasive breast cancer. AB - PURPOSE: Overexpression of HER-2/neu is associated with aggressive diseaseand perhaps with increased risk of breast cancer when presentin benign breast tissue. Breast ductal fluid can be obtained from women by simple nipple aspiration and may be useful for analyzing the microenvironment of the breast. EXPERIMENTAL DESIGN: After obtaining informed consent, we prospectively compared the volume of fluid collected, protein concentration, and level of HER-2/neu expression in nipple aspiration fluid (NAF) samples from both breasts and serum samples in 65 patients with unilateral primary invasive breast cancer (median age, 54 years). HER-2/neu concentrations were determined by immunoassay, with a sensitivity of 0.1 ng/ml. RESULTS: The mean NAF volume obtained and the mean NAF protein concentration were no different in the normal versus the affected breast (62.4 versus 60.4 micro l and 140.9 versus 107.8 mg/ml, respectively). Mean serum HER 2/neu level was 4.36 ng/ml (range, 0-16.8 ng/ml), approximately 50 times less than the mean NAF HER-2/neu level from all patients and all breasts (209.2 ng/ml; range, 1.0-3480.0). NAF HER-2/neu levels were significantly correlated between breasts for each individual patient (r = 0.302; P = 0.038). HER-2/neu overexpressing tumors produced significantly more HER-2/neu in the affected breast (653.6 ng/ml) than in the unaffected breast (101.7 ng/ml) or serum (3.46 ng/ml; P = 0.016). CONCLUSIONS: Nipple aspiration is a noninvasive method for detecting tumor-specific relevant molecular changes from ductal fluid. The presence of high HER-2/neu levels in the ductal systems of breast cancer patients may have clinical implications for monoclonal antibody directed therapy. PMID- 12576425 TI - Increase of regulatory T cells in the peripheral blood of cancer patients. AB - PURPOSE: T cells constitutively expressing both CD4 and CD25are essential for maintenance of self-tolerance and therefore have been referred to as regulatory T cells (Treg). Experimental tumor models in mice revealed that Tregs are potent inhibitors of an antitumor immune response. The current study was designed to determine whether cancer patients exhibit an expanded Treg pool. EXPERIMENTAL DESIGN: The frequency of Tregs in the peripheral blood of 42 patients suffering from epithelial malignancies and from 34 healthy controls was determined by flow cytometry. The immunoregulatory properties of CD4(+)CD25(+) and CD4(+)CD25(-) T cells were characterized by proliferation and suppression assays. Cocultures with natural killer (NK) cells were performed to determine the impact of Tregs on NK mediated cytotoxicity. RESULTS: Patients with epithelial malignancies show an increase of CD4(+)CD25(+) T cells in the peripheral blood with characteristics of Tregs, i.e., they are CD45RA(-), CTLA-4(+), and transforming growth factor beta(+). Notably, CD4(+) T cells from cancer patients are characterized by an impaired proliferative capacity, which is restored to the extend of CD25-depleted CD4(+) T cells from control persons by prior removal of CD25(+) T cells. In contrast to CD4(+)CD25(-) T cells, isolated CD4(+)CD25(+) T cells from cancer patients were anergic towards T cell receptor stimulation. In addition, CD4(+)CD25(+) T cells suppressed the proliferation of CD4(+)CD25(-) T cells. When cultured together with CD56(+) NK-cells, CD4(+)CD25(+) T cells from cancer patients effectively inhibited NK-cell-mediated cytotoxicity. CONCLUSIONS: Thus, we provide evidence of an increased pool of CD4(+)CD25(+) regulatory T cells in the peripheral blood of cancer patients with potent immunosuppressive features. These findings should be considered for the design of immunomodulatory therapies such as dendritic cell vaccination. PMID- 12576426 TI - Predictive value of the EphA2 receptor tyrosine kinase in lung cancer recurrence and survival. AB - PURPOSE: Underestimation of disease severity is a major problem confronting the successful clinical management of non-small cell lung cancer. Recent advances in molecular biological substaging may provide an opportunity to identify those patients with the most aggressive forms of the disease, but there is a continuing need for accurate markers of disease relapse and survival. EXPERIMENTAL DESIGN: In our present study, immunohistochemical analyses of a retrospective database of pathologic specimens were used to demonstrate that the EphA2 receptor kinase is frequently overexpressed in NSCLC. RESULTS: Initial presentation with high levels of EphA2 predicts subsequent survival, overall relapse, and site of relapse. Specifically, high levels of EphA2 in the primary tumor predict brain metastases, whereas low levels of EphA2 relate to disease-free survival or contralateral lung metastasis. CONCLUSIONS: These data suggest that EphA2 may provide a molecular marker to identify and predict patients who have isolated brain metastases. Moreover, the high levels of EphA2 in lung cancer may provide an opportunity for therapeutic targeting. PMID- 12576427 TI - CD40 is expressed on ovarian cancer cells and can be utilized for targeting adenoviruses. AB - PURPOSE: CD40, a member of the tumor necrosis factor receptor superfamily, is widely expressed on various cell types in addition to hematopoietic cells. Recent studies show that CD40 expression is related to several carcinomas, although its role in cancer pathobiology is unknown. In this study, we demonstrate the expression of CD40 on several ovarian carcinoma cell lines and the ability of CD40 to mediate targeted adenoviral infection in vitro. EXPERIMENTAL DESIGN: CD40 expression on ovarian cancer cell lines and clinical patient samples was examined by reverse transcription-PCR and flow cytometry. To study the utilization of CD40 for gene delivery, we precomplexed a luciferase coding adenovirus (Ad), Ad5luc1, with a CD40-targeting molecule (CAR/G28). RESULTS: According to our studies, all of the examined ovarian cancer cell lines are expressing CD40. In addition, mRNA for CD40 was detected in every primary tumor sample, suggesting that CD40 is also expressed in vivo. Compared with nontargeted Ad, gene transfer was increased up to 40-fold in CD40+ cells when CD40-targeted Ad was used. Supporting the relation of targeted system to CD40, increasing the amount of targeting fusion protein results in dose response. Furthermore, blockade of CD40 receptors on cell surface decreases the infectability of CD40+ cells with CD40-targeted virus, indicating the specificity of the targeting system for CD40. CONCLUSIONS: These results suggest that CD40 is present in ovarian cancer cells and can be used for targeted gene delivery in a CAR-independent manner, circumventing the problem of the low expression levels of CAR in various cancer cells. PMID- 12576429 TI - A pilot study of protracted topotecan dosing using a pharmacokinetically guided dosing approach in children with solid tumors. AB - PURPOSE: To assess the use of a pharmacokinetically guided topotecan strategy and evaluate the toxicity of protracted i.v. topotecan in children with recurrent solid tumors. EXPERIMENTAL DESIGN: Fifteen children with measurable relapsed or refractory solid tumors received topotecan i.v. over 30 min 5 days a week for two consecutive weeks. Doses were individualized based on the patient's topotecan systemic clearance to attain a single day topotecan lactone area under the plasma concentration time curve (AUC) of 120-180 ng/ml x h (cohort 1) or 80-120 ng/ml x h (cohort 2). Clinical responses and toxicity were assessed by standard criteria. RESULTS: Twenty-nine courses of topotecan were administered, 11 in cohort 1 and 18 in cohort 2. The median topotecan dosages required to achieve the target AUCs for cohorts 1 and 2 were 4 mg/m(2) (range, 2.6-6) and 3 mg/m(2) (range, 2.6-4.2), respectively. The intersubject variance for topotecan clearance exceeded the intrasubject variance by 2-fold. With the pharmacokinetic targeting approach, we observed that 78% (46 of 59) of the measured AUC values were within the target range. The median number of days to an absolute neutrophil count >/=500/mm(3) was similar between the two cohorts; however, febrile neutropenia and serious infections limited our ability to deliver drug dosages needed to secure the higher systemic exposure (cohort 1). Five partial responses were observed. CONCLUSION: Protracted topotecan dosing using a pharmacokinetic strategy was possible in this heavily pretreated group of children. PMID- 12576428 TI - Alpha1 acid glycoprotein binds to imatinib (STI571) and substantially alters its pharmacokinetics in chronic myeloid leukemia patients. AB - PURPOSE: Imatinib (Glivec) is a potent inhibitor of bcr/abl, an oncogenic fusion protein that causes chronic myelogenous leukemia (CML). alpha1 acid glycoprotein (AGP) binds to imatinib with high affinity and inhibits imatinib activity in vitro and in vivo in an animal model. A pharmacokinetics analysis of imatinib was undertaken in CML patients. EXPERIMENTAL DESIGN: Imatinib plasma concentrations were measured in 19 CML patients treated with imatinib (400 or 600 mg/day). Five patients received a concomitant short-term course of clindamycin (CLI). RESULTS: A positive correlation between AGP and imatinib plasma levels was observed. CLI administration decreased imatinib plasma concentrations, evaluated as area under the curve (AUC) and peak concentrations (C(max)). The effects of a bolus of CLI was studied in three patients on imatinib 23 h after the last imatinib dose. Within 5-10 min in three of three cases, CLI caused a decrease in imatinib plasma concentrations of 2.6-, 2.7-, and 4.7-fold, respectively. In vitro experiments using fresh blasts from CML patients showed that AGP, at concentrations observed in the patients, decreased imatinib intracellular concentrations up to 10 times and blocked imatinib activity. The incubation with CLI restored imatinib intracellular concentrations and biological activity. CONCLUSION: AGP exerts significant effects of the pharmacokinetics, plasma concentrations, and intracellular distribution of imatinib in CML patients; these data indicate that plasma imatinib levels represent unreliable indicators of the cellular concentrations of this molecule. PMID- 12576430 TI - Enzyme-linked immunospot, cytokine flow cytometry, and tetramers in the detection of T-cell responses to a dendritic cell-based multipeptide vaccine in patients with melanoma. AB - A new generation of single-cell assays for measuring the frequency of peptide specific T lymphocytes in cellular populations has become widely available. These assays, enzyme-linked immunospot (ELISPOT), cytokine flow cytometry, and tetramer binding, are used frequently for monitoring of cancer patient responses to vaccination therapies. We concomitantly used these three assays to determine the frequency of CD8(+) T cells in the circulation of 8 patients with metastatic melanoma who had received a multiepitope peptide/dendritic cell-based vaccine. Using peripheral blood mononuclear cells harvested before and a week after the last of four vaccines, we observed that the three assays detected a substantially different frequency of peptide-specific CD8(+) T cells. The tetramer assay consistently detected the highest numbers of peptide-specific CD8(+) T cells, followed by cytokine flow cytometry and then ELISPOT. There was no significant concordance among the assays in measuring the numbers of CD8(+) T cells specific for each of the peptides in the peripheral circulation of the patient. No significant pre- to postvaccine changes in the number of CD8(+) T cells specific for any of the peptides were observed. Thus, the dendritic cell-based vaccine was observed not to augment immune responses to the peptides in the patients. Because of a low frequency of the peptide-specific T cells in the peripheral circulation of the patients, these sensitive single-cell assays were used to measure values at the lower limit of detection. For this and other reasons, including the issues of tetramer specificity and ELISPOT sensitivity, caution in interpretation and serious attention to quality control are needed in monitoring of immune responses to anticancer vaccines. PMID- 12576431 TI - Increased 99mTc-sestamibi accumulation in normal liver and drug-resistant tumors after the administration of the glycoprotein inhibitor, XR9576. AB - 99mTc-sestamibi, a substrate of the multidrug transporter P-glycoprotein (Pgp), has been used as a functional imaging agent for the multidrug resistance-1 (MDR1) phenotype. In vitro, retention of (99m)Tc-sestamibi by cells that overexpress Pgp can be enhanced by the addition of Pgp inhibitors. XR9576 (Tariquidar) is a potent and selective noncompetitive inhibitor of Pgp that is active at 25-80 nM. A Phase I trial of XR9576 in combination with vinorelbine (Navelbine) was conducted in 26 patients with metastatic cancers. A (99m)Tc-sestamibi scan was obtained at baseline followed 48-96 h later by a second scan 1-3 h after the administration of XR9576. Time activity curves and areas under the curves (AUCs) were obtained for tumor, liver, lung, and heart, and tissue:heart AUC ratios were calculated. XR9576 enhanced (99m)Tc-sestamibi accumulation and retention in the liver of all but two patients with a mean change of +128%. Furthermore, in 13 of 17 patients with tumor masses visible by (99m)Tc-sestamibi, the tumor:heart (99m)Tc-sestamibi AUC(0-3 h) increased after the administration of XR9576, with increases of 36-263% seen in 8 patients. We conclude that in vivo administration of XR9576 inhibits (99m)Tc-sestamibi efflux in both the normal liver and in drug resistant tumors. This study provides convincing evidence of the existence of XR9576-inhibitable (99m)Tc-sestamibi efflux in a large fraction of drug resistant tumors. One can predict that efflux of Pgp substrates also occurs in these tumors. XR9576 provides an efficient way to inhibit this efflux and offers the potential to increase drug exposure in human cancer. PMID- 12576432 TI - Vaccine-induced CD8+ T-cell responses to MAGE-3 correlate with clinical outcome in patients with melanoma. AB - PURPOSE: Vaccine-induced antitumor CD8+ T-cell responses are believed to play an important role in increasing resistance to melanoma. The following study was conducted to examine whether these responses are associated with improved clinical outcome in melanoma vaccine-treated patients. EXPERIMENTAL DESIGN: We measured CD8+ T-cell responses to gp100, MART-1, MAGE-3, and tyrosinase by enzyme linked immunospot assay in peripheral blood of 131 HLA-A*01- or HLA-A*02-positive melanoma patients before and after immunization to a polyvalent, shed antigen, melanoma vaccine, and correlated the results with clinical outcome. RESULTS: Fifty-six percent of patients had a vaccine-induced CD8+ T-cell response to at least one of the four antigens. Recurrences were significantly reduced in patients with vaccine-induced responses to MAGE-3 (hazard ratio, 0.42; 95% confidence interval, 0.18-0.99; P = 0.03) by the Cox proportional hazard model but were unrelated to responses to the other three antigens. Patients with a preexisting response to any of the four antigens were significantly more likely to have a further vaccine-boosted response to that same antigen (P < 0.0001 0.036). CONCLUSIONS: There was a correlation between vaccine-induced CD8+ T-cell responses to melanoma-associated antigens and improved clinical outcome, but the correlation depended on the antigen against which the response is directed. The only significant correlation was with responses to MAGE-3. PMID- 12576433 TI - Phase I evaluation of prolonged-infusion gemcitabine with fludarabine for relapsed or refractory acute myelogenous leukemia. AB - PURPOSE: The purpose of this study was to determine the maximum tolerated duration of infusion of gemcitabine at 10 mg/m(2)/min in combination with fludarabine at 25 mg/m(2) daily for 5 days in the treatment of relapsed or refractory acute myelogenous leukemia. EXPERIMENTAL DESIGN: Eighteen patients with relapsed or refractory acute myelogenous leukemia were enrolled. The median age was 54.5 years (range, 21-80 years). Patients received a 30-min infusion of fludarabine at 25 mg/m(2) daily for 5 days. i.v. gemcitabine was given as a single infusion at 10 mg/m(2)/min with the duration adjusted following a modified continuous reassessment method. RESULTS: After 18 patients, the maximum recommended duration of infusion of gemcitabine in combination with fludarabine was selected as a 15-h infusion given at 10 mg/m(2)/min (9,000 mg/m(2)). Severe stomatitis or esophagitis was the most common nonhematological dose-limiting toxicity. Myelosuppression was universal. Febrile neutropenia was common, and 3 of 18 (17%) patients developed bacteremia. Occasional nausea, vomiting, or diarrhea was also reported. There were three complete responses and two partial responses for an overall response rate of 28%. CONCLUSIONS: Prolonged-infusion gemcitabine at a fixed dose rate of 10 mg/m(2)/min for 15 h with 25 mg/m(2)/day fludarabine for 5 days is a tolerable induction regimen for relapsed or refractory leukemia. Stomatitis, esophagitis, febrile neutropenia, and myelosuppression should be anticipated; however, this regimen may be beneficial in patients with relapsed or refractory leukemia. PMID- 12576434 TI - Activation of human melanoma reactive CD8+ T cells by vaccination with an immunogenic peptide analog derived from Melan-A/melanoma antigen recognized by T cells-1. AB - PURPOSE: As compared with natural tumor peptide sequences, carefully selected analog peptides may be more immunogenic and thus better suited for vaccination. However, T cells in vivo activated by such altered analog peptides may not necessarily be tumor specific because sequence and structure of peptide analogs differ from corresponding natural peptides. EXPERIMENTAL DESIGN: Three melanoma patients were immunized with a Melan-A peptide analog that binds more strongly to HLA-A*0201 and is more immunogenic than the natural sequence. This peptide was injected together with a saponin-based adjuvant, followed by surgical removal of lymph node(s) draining the site of vaccination. RESULTS: Ex vivo analysis of vaccine site draining lymph nodes revealed antigen-specific CD8+ T cells, which had differentiated to memory cells. In vitro, these cells showed accelerated proliferation upon peptide stimulation. Nearly all (16 of 17) of Melan-A-specific CD8+ T-cell clones generated from these lymph nodes efficiently killed melanoma cells. CONCLUSIONS: Patient immunization with the analog peptide leads to in vivo activation of T cells that were specific for the natural tumor antigen, demonstrating the usefulness of the analog peptide for melanoma immunotherapy. PMID- 12576435 TI - Mechanisms of peritoneal metastasis after operation for non-serosa-invasive gastric carcinoma: an ultrarapid detection system for intraperitoneal free cancer cells and a prophylactic strategy for peritoneal metastasis. AB - PURPOSE: This aims of this study are to establish an ultra-rapid quantitative reverse transcription-PCR (RT-PCR) protocol that enables the diagnosis of i.p. cancer spread during operation, to reveal the mechanisms of peritoneal metastasis from non-serosa-invasive gastric carcinoma, and to evaluate the effect of the extensive intraoperative peritoneal lavage (EIPL) using the ultra-rapid quantitative RT-PCR as a prophylactic strategy for peritoneal metastasis. EXPERIMENTAL DESIGN: Peritoneal lavage samples from 63 patients with non-serosa invasive gastric carcinoma were obtained at laparotomy and immediately after lymph node dissection. To identify the free cancer cells in the samples, carcinoembryonic antigen- and cytokeratin 20-specific RT-PCRs were performed using the LightCycler method in combination with an automated mRNA extractor. In addition, EIPL was performed in five cases with serosa-invasive gastric carcinoma, and its efficacy was evaluated by the ultra-rapid quantitative RT-PCR protocol. RESULTS: The method enabled us to complete the detection of cancer cells within approximately 70 min. Both the carcinoembryonic antigen and cytokeratin 20 mRNA in i.p. lavages after lymph node dissection were identified in three (14.3%), four (26.7%), and six (46.2%) patients with submucosal, muscularis propria, and subserosal tumors, respectively. Lymph node metastasis was the independent predictor of the existence of i.p. free cancer cells. The ultra-rapid quantitative RT-PCR demonstrated that EIPL reduced free cancer cells from 3.8 x 10(5) +/- 1.4 x 10(5) cells to 2.8 +/- 1.5 cells/100 ml lavage after six to eight washes, and they disappeared after seventh to ninth wash. CONCLUSIONS: The present study proved that lymph node dissection opened lymphatic channels and spread viable cancer cells into the peritoneal cavity. It is suggested that the combination of the novel detection system with the intraoperative therapy of EIPL can be a useful prophylactic strategy for peritoneal metastasis from gastric carcinoma. PMID- 12576436 TI - Weekly docetaxel as neoadjuvant chemotherapy for stage II and III breast cancer: efficacy and correlation with biological markers in a phase II, multicenter study. AB - PURPOSE: This is one of the first reports of weekly docetaxel (Taxotere) in the neoadjuvant treatment of stage II and III breast cancer. We evaluated docetaxel's efficacy and safety and analyzed correlations between response and the expression of c-erbB2, ER status, and Ki-67 labeling index. EXPERIMENTAL DESIGN: Patients with previously untreated, stage II and III breast cancer were entered into the study. Docetaxel (40 mg/m(2)) was given i.v. once weekly for the first 6 weeks of an 8-week cycle for 2 cycles. RESULTS: A total of 56 patients were evaluated by intention-to-treat analysis for efficacy and safety. The overall clinical response rate was 68% (complete and partial response, 29 and 39%, respectively). Nine patients (16%) achieved a pathological complete response. There was no correlation between response to docetaxel and the expression of molecular markers, however, the majority of the pathological complete responses were observed in patients with c-erbB2-negative tumors. Nonhematological toxicity was more common than hematological toxicity, with alopecia and asthenia the most frequently reported adverse events (89 and 77% of patients, respectively). Severe hematological toxicity was rare. CONCLUSIONS: Weekly docetaxel appears to be very effective in the neoadjuvant setting. A high pathological response rate was achieved with tolerable toxicity. PMID- 12576437 TI - Phase II clinical trial of intralesional administration of the oncolytic adenovirus ONYX-015 in patients with hepatobiliary tumors with correlative p53 studies. AB - PURPOSE: ONYX-015 is a genetically modified adenovirus with a deletion of the E1B early gene and is therefore designed to replicate preferentially in p53-mutated cells. A Phase II trial of intralesional ONYX-015 was conducted in patients with hepatobiliary tumors to determine the safety and efficacy of such a treatment. EXPERIMENTAL DESIGN: All patients had biopsy-proven, measurable tumors of the liver, gall bladder, or bile ducts that were beyond the scope of surgical resection. Patients received intralesional injections of ONYX-015 at either 6 x 10(9) or 1 x 10(10) plaque-forming units/lesion up to a total dose of 3 x 10(10) plaque-forming units, and i.p. injections were allowed in patients with malignant ascites. The status of p53 was assessed by immunohistochemistry or Affymetrix GeneChip microarray analysis. Studies were conducted for viral shedding and for the presence of antiadenoviral antibodies before and after the injection of ONYX 015. Patients were assessed for response and toxicity. RESULTS: Twenty patients were enrolled, and 19 patients were eligible. Half of the patients had primary bile duct carcinomas. Serious toxicities (> grade 2) were uncommon and included hepatic toxicity (three patients), anemia (one patient), infection (one patient), and cardiac toxicity (one patient, atrial fibrillation). Sixteen patients were evaluable for response. Among these evaluable patients, 1 of 16 (6.3%) had a partial response, 1 of 16 (6.3%) had prolonged disease stabilization (49 weeks), and 8 of 16 (50%) had a >50% reduction in tumor markers. Of the 19 eligible patients, 18 (94.7%) had specimens available for p53 analysis. Fifteen of these 18 patients (83.3%) had evidence of p53 mutation by one or both methods, although the methods correlated poorly. Viral shedding was confined to bile (two of two patients) and ascites (four of four patients). Pretreatment adenoviral antibodies were present in 14 of 14 patients and increased by 33.2% after ONYX-015 treatment. CONCLUSIONS: Intralesional treatment with ONYX-015 in patients with hepatobiliary tumors is safe and well tolerated, and some patients had evidence of an anticancer effect. The high incidence of p53 mutations in these tumors makes this a logical population in which to test this therapy but precludes definitive evaluation about the necessity of a p53 mutation for ONYX-015 clinical activity. PMID- 12576438 TI - Phase I clinical and pharmacologic study of weekly cisplatin and irinotecan combined with amifostine for refractory solid tumors. AB - PURPOSE: This Phase I study was designed primarily to determine the maximum tolerated dose (MTD) and dose-limiting toxicities (DLTs) of irinotecan and cisplatin with and without amifostine in children with refractory solid tumors. PATIENTS AND METHODS: Cisplatin, at a fixed dose of 30 mg/m(2), and escalating doses of irinotecan (starting dose, 40 mg/m(2)) were administered weekly for four consecutive weeks, every 6 weeks. After the MTD of irinotecan plus cisplatin was determined, additional cohorts of patients were enrolled with amifostine (825 mg/m(2)) support. Leukocyte DNA-platinum adducts and pharmacokinetics of cisplatin and WR-1065 (amifostine-active metabolite) were also determined. RESULTS: Twenty-four patients received 43 courses of therapy. The MTD for irinotecan administered in combination with cisplatin (30 mg/m(2)) was 50 mg/m(2). The DLTs of this combination were neutropenia and thrombocytopenia. With the addition of amifostine, at an irinotecan dose of 65 mg/m(2) and cisplatin dose of 30 mg/m(2), the DLT was hypocalcemia. Although no objective responses were observed, six patients received at least three courses of therapy. The amounts of platinum adducts (mean +/- SD) were 10 +/- 20 molecules/10(6) nucleotides. The maximum plasma concentrations (C(max)) for free cisplatin and WR 1065 were 4.5 +/- 1.6 micro M and approximately 89 +/- 10 micro M, respectively. The half-life (t(1/2)) for free plasma cisplatin was 25.4 +/- 5.4 min. The initial t(1/2) for plasma WR-1065 was approximately 7 min and terminal t(1/2) approximately 24 min. CONCLUSION: The combination of cisplatin and irinotecan administered weekly for 4 weeks in children with refractory cancer is well tolerated. Amifostine offers some myeloprotection, likely permitting >/=30% dose escalation for irinotecan, when administered in a combination regimen with cisplatin. However, effective antiemetics and calcium supplementation are necessary with the use of amifostine. Further escalation of irinotecan dosing, using these precautions for amifostine administration, may be possible. PMID- 12576439 TI - High-dose methotrexate for intraocular lymphoma. AB - PURPOSE: Intraocular lymphoma (IOL) frequently coexists with primary central nervous system lymphoma(PCNSL). We sought to determine the efficacy of high-dose methotrexate (MTX) alone in patients with IOL. We also sought to determine whether micromolar levels of MTX could be achieved in the aqueous and vitreous humor of the eye after i.v. administration of the drug. EXPERIMENTAL DESIGN: Nine patients with concurrent PCNSL and IOL or isolated IOL were treated with MTX alone. All patients were treated with i.v. 8 g/m(2) MTX. MTX concentrations in serum, aqueous humor, and vitreous humor were obtained in seven of nine patients with IOL and in one additional patient with PCNSL but no evidence of IOL. RESULTS: Micromolar concentrations of MTX were present in both ocular chambers 4 h after completion of the infusion in eight of eight patients. Levels of MTX were lower in the vitreous humor compared with the aqueous humor in five of six patients in whom both chambers were assayed. Initial response of IOL to MTX was demonstrated by seven of nine patients (six complete responses and one partial response), whereas two patients had persistent IOL despite achievement of micromolar concentrations of MTX. In the patients with concurrent PCNSL and IOL, seven of seven had complete responses in the brain after treatment with MTX. Three of seven patients with initial response of IOL experienced relapse in the eye requiring orbital radiation, and four of nine patients had sustained response of IOL to MTX. CONCLUSIONS: A subset of patients with IOL may experience sustained remission when treated with high-dose i.v. MTX alone. Although micromolar MTX concentrations are present in the eye 4 h after infusion, the lower concentration achieved in vitreous humor may contribute to persistence of IOL. PMID- 12576440 TI - Prognostic significance of vascular endothelial growth factor D in breast carcinoma with long-term follow-up. AB - PURPOSE: Expression of angiogenic and lymphangiogenic factors by tumors may influence the route of metastatic spread. The angiogenic factor vascular endothelial growth D (VEGF-D) is implicated in the development of lymphatic vessels and promotion of lymphatic metastases. The purpose of this study is to determine whether VEGF-D correlates with lymph node metastasis or prognosis. EXPERIMENTAL DESIGN: We assessed VEGF-D expression using immunohistochemistry in 105 invasive breast carcinomas with long-term follow-up. The relationship among VEGF-D expression, lymph node status, and other established clinicopathological parameters was assessed. Whether VEGF-D expression plays prognostic role in breast cancer was also investigated. RESULTS: VEGF-D expression was identified in 86 cases (81.9%). Positive VEGF-D was significantly correlated with lymph node metastasis (P = 0.0238) and high c-erbB-2 expression (P = 0.0211). Survival curves determined by the Kaplan-Meier method and univariate analysis demonstrated that positive VEGF-D was associated with both disease-free survival (P = 0.0023) and overall survival (P = 0.0222). In multivariate analysis using the Cox regression model, positive emerged as an independent indicator for disease-free survival (P = 0.0452). CONCLUSIONS: VEGF-D expression is associated with lymph node metastasis and may be a novel prognostic factor in breast cancer. VEGF-D may be useful in the treatment of breast cancer as a decision-making biomarker for aggressive treatment after operation. PMID- 12576441 TI - Immunohistochemical expression of CD23 and CD40 may identify prognostically favorable subgroups of diffuse large B-cell lymphoma: a Nordic Lymphoma Group Study. AB - PURPOSE: In search for subgroups of diffuse large B-cell lymphoma (DLBCL) with different histogenetic origin and prognosis, as has been described by gene expression profiling, we examined tumor specimens from 125 patients with DLBCL, uniformly treated by either cyclophosphamideAdriamycin-vincristine-prednisone or methotrexate, doxorubicin, cyclophosphamide, vincristine, prednisone, and bleomycin in a multicenter trial set by the Nordic Lymphoma Group 1989-1994. EXPERIMENTAL DESIGN: Bcl-6, CD10, and CD40 were chosen as markers for a germinal center phenotype, CD23 as a marker of pre/early germinal center origin, and CD138 as a marker for postgerminal center origin. In addition, expression of the apoptotic regulators bcl-2 and bax was analyzed. Immunohistochemical analysis was performed using the EnVision method. RESULTS: CD10 was positive in 51%, bcl-6 in 97%, and CD138 only in 2% of the cases. No prognostic conclusions could be drawn from analysis of these factors. CD40 was positive in 76% of the cases. This group was associated with superior time to treatment failure (P = 0.027) and overall survival (OS; P = 0.0068). By Cox regression analysis, positivity for CD40 was shown to be a prognostic factor for OS, independent of International Prognostic Index. CD23 was positive in 16% of the cases (all CD5 negative and all CD40 positive). This group showed a strong tendency for better OS (P = 0.033). CD40 expression correlated with bax but not with bcl-2 expression. CONCLUSIONS: CD23 and CD40 expression seems to be prognostically favorable in DLBCL. This may be secondary to a germinal center origin or attributable to increased apoptosis via induction of bax and/or enhanced T-cell interaction, resulting in improved autologous tumor response. Confirmatory studies are necessary. PMID- 12576442 TI - Up-regulation of tumor interleukin-8 expression by infiltrating macrophages: its correlation with tumor angiogenesis and patient survival in non-small cell lung cancer. AB - PURPOSE: To evaluate the interaction between tumor-infiltrating macrophages and cancer cells and its effect on the expression of a potent angiogenic factor, interleukin-8 (IL-8), tumor angiogenesis, and patient outcome in non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: We measured tumor IL-8 mRNA expression (by real-time quantitative reverse transcription-PCR), intratumor microvessel counts, and tumor-infiltrating macrophage density (by immunohistochemical staining) in 35 NSCLC surgical specimens and correlated with the patient's clinical outcome. We then investigated the interaction between macrophages (cell line THP-1) and six different human cancer cell lines (four NSCLCs, one osteosarcoma, and one hepatoma) and its effect on IL-8 mRNA expression using a macrophage/cancer cell coculture system, IL-8 mRNA expression in lung cancer cells, and macrophages being measured separately after coculture in the presence or absence of six anti-inflammatory agents, i.e., pentoxifylline, aspirin, indomethacin, dexamethasone, celecoxib (a selective cyclooxygenase-2 inhibitor), and pyrrolidine dithiocarbamate, a specific nuclear factor kappaB (NF-kappaB) inhibitor. NF-kappaB transcriptional activity and protein levels were measured by reporter gene assay and Western blot. RESULTS: The tumor-infiltrating macrophage density correlated significantly and positively with tumor IL-8 mRNA expression and intratumor microvessel counts and significantly and negatively with patient survival. In addition, after cell-cell interaction in cancer cell:macrophage cocultures, marked IL-8 mRNA expression was induced in lung cancer cells (approximately 270-fold) and, to a lesser degree, in macrophages (4.5-fold). The increase in IL-8 mRNA expression correlated with the in vitro metastatic potential of the cancer cells. All six anti-inflammatory agents suppressed induction of IL-8 mRNA expression in lung cancer cells by >90%, four (pentoxifylline, celecoxib, pyrrolidine dithiocarbamate, and dexamethasone) having a dose-dependent effect. NF-kappaB transcriptional regulation and protein levels were simultaneously increased in the nuclei of cancer cells in macrophage/cancer cell cocultures, this effect also being suppressed by all six anti-inflammatory agents. CONCLUSIONS: The interaction between infiltrating macrophages and cancer cells up-regulates IL-8 mRNA expression, especially in the cancer cells; this may contribute greatly to the increased tumor angiogenesis and adverse outcome in NSCLC patients with a high density of tumor-infiltrating macrophages. Anti-inflammatory agents can suppress the induction of IL-8 mRNA expression seen in lung cancer cells after coculture with macrophages, and this suppression is mediated, in part, through the NF-kappaB pathway. PMID- 12576443 TI - Caspase 3 in breast cancer. AB - PURPOSE: An inability to undergo apoptosis is widely thought to contribute to both tumorigenesis and tumor progression. One of the key mediators of apoptosis is the thiol protease caspase 3. In this investigation, caspase 3 mRNA and protein expression in breast cancer was examined. EXPERIMENTAL DESIGN: Caspase 3 was measured at the mRNA level using reverse transcription-PCR and at the protein level using both Western blotting and activity assays. Levels of apoptosis were determined using an ELISA, which detects nucleosomes released during DNA fragmentation. RESULTS: Relative levels of caspase 3 mRNA were similar in breast carcinomas (n = 103), fibroadenomas (n = 25), and normal breast tissues (n = 6). However, levels of both the precursor and active forms of caspase 3 were significantly higher in carcinomas compared with both fibroadenomas (P = 0.0188) and normal breast tissues (P = 0.0002). Levels of apoptosis were also highest in the carcinomas and correlated significantly with active caspase 3 levels (r = 0.481; P = 0.0003). In the carcinomas, expression of caspase 3 showed no significant relationship with either tumor size, tumor grade, nodal status, or steroid receptor status but was significantly higher in ductal carcinomas than in lobular carcinomas (P = 0.0188). CONCLUSIONS: We conclude that rates of apoptosis as measured by both caspase 3 activation and nucleosome release are higher in breast cancer than in nonmalignant breast tissue. This finding would appear to conflict with the widely held belief that apoptosis is reduced in malignancy. The proliferation:apoptosis ratio, however, may be higher in carcinomas than in the corresponding normal tissue. PMID- 12576444 TI - Methylenetetrahydrofolate reductase 677 C-->T polymorphism and risk of proximal colon cancer in north Italy. AB - PURPOSE: Gene silencing by hypermethylation plays an important role in proximal colon carcinogenesis. Conversely, DNA hypomethylation has been associated with distal colon cancer (CLC). Methylenetetrahydrofolate reductase (MTHFR) catalyzes the conversion of 5',10'-methylenetetrahydrofolate to 5'-methyl tetrahydrofolate, which serves as methyl donor in the remethylation of homocysteine to methionine. A common MTHFR 677 C-->T polymorphism is characterized by reduced catalytic activity, which affects methionine synthesis and DNA methylation. The aim of the study was to investigate the role of MTHFR 677 C-->T gene polymorphism in the tumorigenesis of proximal and distal CLC in a monoinstitutional group of patients in North Italy. EXPERIMENTAL DESIGN: One-hundred thirty four consecutive proximal and 142 consecutive distal CLC patients, and 279 control subjects without cancer were genotyped for MTHFR using PCR-restriction fragment-length polymorphism analysis. RESULTS: The prevalence of the 677 TT genotype was significantly (P = 0.005) lower in patients with proximal tumors (10 of 134, 7%) than in subjects with distal tumors (28 of 142, 20%). Case/control approach indicated that individuals homozygous for the 677 TT allele had a significantly reduced risk (2.8-fold) (adjusted odds ratio, 0.36; 95% confidence intervals, 0.14-0.91) of developing proximal CLC compared with those harboring the wild-type or heterozygous genotype (677 CC or 677 CT). No significant association between CLC risk and TT genotype was observed in patients with distal tumors (odds ratio, 1.01; 95% confidence interval, 0.48-2.14). CONCLUSIONS: Our findings support a role for MTHFR 677 TT genotype in reducing proximal CLC risk in North Italy. PMID- 12576445 TI - Serum antibodies to lipophilin B detected in late stage breast cancer patients. AB - Lipophilin B mRNA is overexpressed in approximately 70% of breast tumors and shows a high degree of correlation with the mRNA expression profile of mammaglobin. This is further supported by the recent finding that, like other members of the secretoglobulin-uteroglobin family, mammaglobin and lipophilin B form a heteroduplex. The studies described show that there are pre-existing antibodies to lipophilin B peptide in the sera of breast cancer patients with different stages and grade of tumor and that this response is different from that seen to recombinant mammaglobin and native mammaglobin-lipophilin B complex. The highest titers were observed in later stage tumors. In addition, low levels of antibody were also seen in some patients with prostate and ovarian cancers, consistent with lipophilin B mRNA expression in these tumors at lower abundance than in breast tumors. In contrast, lipophilin B antibodies were absent in 20 healthy donor sera and 30 lung cancer sera. A polymorphism identified in Lipophilin B did not appear to influence human sera reactivity. The data indicate that humoral immune responses to lipophilin B may serve as a diagnostic indicator, particularly for breast cancer. PMID- 12576446 TI - Quantitative molecular detection of minimal residual head and neck cancer in lymph node aspirates. AB - PURPOSE: Staging of the clinically N(0) neck in patients with head and neck squamous cell carcinoma (HNSCC) using ultrasound-guided, fine needle aspiration cytology (USgFNAC) has a false-negative rate of approximately 20% that might be caused by inaccurate cytology. Molecular analysis of aspirate residues might reduce the false-negative rate, and we therefore set up a quantitative reverse transcription-PCR (Q-RT-PCR) assay based on TaqMan technology using the squamous cell-specific antigen E48 (Ly-6D) as molecular marker. EXPERIMENTAL DESIGN: The detection limit of the assay was determined in reconstruction experiments. The sensitivity of the assay was tested on cytological tumor-positive aspirate residues and the specificity on lymph node aspirate residues of noncancer controls. Subsequently, 235 lymph node aspirate residues of 64 HNSCC patients staged with USgFNAC were examined for the presence of E48 mRNA. E48 Q-RT-PCR results of the aspirated lymph nodes were compared with cytology and clinical outcome. RESULTS: The detection limit of E48 Q-RT-PCR was a single tumor cell in a background of 10(6) peripheral blood mononuclear cells. From the 41 aspirates that were not evaluable at cytology, 24 (59%) could be diagnosed with E48 Q-RT PCR. In the 191 aspirates that were tumor negative or not evaluable at cytology, 8 samples from 6 patients were E48 positive. These results were confirmed by histology or clinical outcome in 3 of 6 patients. E48 Q-RT-PCR showed an increase in sensitivity from 56 to 67% and an increase in frequency of reached diagnosis from 97 to 100% compared with cytology. The specificity decreased from 100 to 92%. CONCLUSIONS: Real-time E48 Q-RT-PCR is an accurate technique for squamous cell detection in lymph node aspirates of HNSCC patients. The assay shows an increase in sensitivity and frequency of reached diagnosis compared with cytology. The test could be implemented routinely in USgFNAC to diagnose cases for which cytological examination is not conclusive. PMID- 12576447 TI - High expression of tumor necrosis factor-related apoptosis-inducing ligand is associated with favorable ovarian cancer survival. AB - PURPOSE: The molecular determinants of survival in ovarian cancer are poorly understood. Using expression microarrays, we recently found that high expression of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene is associated with prolonged survival in advanced ovarian cancer. TRAIL has also been shown to synergize with chemotherapeutic agents to induce apoptosis in ovarian cancer cell lines. We therefore sought to confirm the association between TRAIL expression and survival in a larger group of women with ovarian cancer. EXPERIMENTAL DESIGN: TRAIL expression was measured using quantitative real-time PCR in 120 epithelial ovarian cancers (11 stage I/II, 109 stage III/IV) and 8 normal ovarian surface epithelial samples. RESULTS: Ovarian cancers demonstrated 10-fold higher mean TRAIL expression than normal ovarian epithelial samples (P < 0.001). Among ovarian cancers, high TRAIL expression was associated with prolonged survival and was 2.2-fold higher in cancers from patients who lived more than 5 years compared with patients who died within 1 year (P = 0.03). CONCLUSIONS: TRAIL expression is higher in ovarian cancers relative to normal ovarian epithelium. High TRAIL expression is associated with favorable ovarian cancer survival, which may be attributable to increased chemosensitivity of cancers that express the most TRAIL. The use of TRAIL to enhance sensitivity of ovarian cancers to therapy represents an appealing molecular therapeutic strategy worthy of further investigation. PMID- 12576448 TI - Molecular determinants of treatment response in human germ cell tumors. AB - PURPOSE: Germ cell tumors (GCTs) are highly sensitive to cisplatin-based chemotherapy. This feature is unexplained, as is the intrinsic chemotherapy resistance of mature teratomas and the resistant phenotype of a minority of refractory GCTs. Various cellular pathways may influence the efficacy of chemotherapy. Their impact has not been investigated in a comprehensive study of tumor samples from clinically defined subgroups of GCT patients. EXPERIMENTAL DESIGN: We investigated proteins involved in regulation of apoptosis (p53, BAX, BCL-2, and BCL-X(L)), cell cycle control [p21 and retinoblastoma protein (RB)], and drug export and inactivation [P-glycoprotein, multidrug resistance-associated protein (MRP) 1, MRP2, breast cancer resistance protein, lung resistance protein, metallothionein, and glutathione S-transferase pi] immunohistochemically in samples of unselected GCT patients (n = 20), patients with advanced metastatic disease in continuous remission after first-line chemotherapy (n = 12), and chemotherapy-refractory patients (n = 24). Mature teratoma components (n = 10) within tumor samples from all groups were analyzed separately. The apoptotic index was studied by terminal deoxynucleotidyl transferase-mediated nick end labeling assay. RESULTS: Invasive GCTs of all groups showed a correlation between wild-type p53 and apoptotic index (r(s) = 0.66; P < 0.001). The levels of the antiapoptotic proteins BCL-2 and BCL-X(L) were generally low. p21 was hardly detectable and did not correlate with p53 (r(s) = 0.29; P = 0.07). No significant differences among the three patient groups were identified regarding any of the investigated parameters (all Ps were >0.08), even though only individual samples from chemotherapy-resistant cases showed a strong staining for MRP2 and GSTpi. In contrast to other components, mature teratomas showed an intense p21 and RB staining and were mostly positive for MRP2, lung resistance protein, and GSTpi. CONCLUSIONS: Our results indicate a multifactorial basis for the chemosensitivity of GCTs with lack of transporters for cisplatin, of antiapoptotic BCL-2 family members, of p21 induction by p53, and of RB and an intact apoptotic cascade downstream of p53. These findings suggest a preference for apoptosis over cell cycle arrest after up-regulation of p53. None of the examined parameters offers a general explanation for the chemotherapy-resistant phenotype of refractory tumors. The up-regulation of various factors interfering with chemotherapy efficacy and ability for a p21-induced cell cycle arrest may explain the intrinsic chemotherapy resistance of mature teratomas. PMID- 12576449 TI - Increased plasma ghrelin level in lung cancer cachexia. AB - PURPOSE: Ghrelin, a novel growth hormone-releasing peptide,has been shown to cause a positive energy balance by stimulating food intake and inducing adiposity. We sought to investigate the pathophysiology of ghrelin in cachexia associated with lung cancer. EXPERIMENTAL DESIGN: Plasma ghrelin level was measured in 43 patients with lung cancer and 21 control subjects. Patients with lung cancer were divided into two groups: patients with cachexia (n = 21) and those without cachexia (n = 22). RESULTS: Plasma ghrelin level did not significantly differ between all patients with lung cancer and controls (157 +/- 10 versus 132 +/- 8 fmol/ml, P = 0.1). However, plasma ghrelin level was significantly higher in patients with cachexia than in those without cachexia (180 +/- 17 versus 135 +/- 10 fmol/ml, P = 0.011). Furthermore, plasma ghrelin level increased significantly in patients with decreased food intake after chemotherapy (from 136 +/- 11 fmol/ml to 170 +/- 16 fmol/ml on day 8, 179 +/- 20 fmol/ml on day 21 after start of chemotherapy), although plasma ghrelin level did not significantly change in those without decreased food intake. CONCLUSIONS: Baseline plasma ghrelin level was elevated in cachectic patients with lung cancer, and follow-up plasma ghrelin level increased in patients with anorexia after chemotherapy. Considering the positive energy effects induced by ghrelin, increased ghrelin may represent a compensatory mechanism under catabolic-anabolic imbalance in cachectic patients with lung cancer. PMID- 12576450 TI - Level of Id-1 protein expression correlates with poor differentiation, enhanced malignant potential, and more aggressive clinical behavior of epithelial ovarian tumors. AB - PURPOSE: Id (inhibitor of differentiation/DNA binding) -1 is involved in neoangiogenesis, it antagonizes basic helix-loop-helix proteins, inhibits differentiation, and enhances cell proliferation. Aim of this study was to investigate Id-1 protein expression in epithelial ovarian tumors and its clinical relevance in ovarian cancer. EXPERIMENTAL DESIGN: We have investigated Id-1 expression by reverse transcription-PCR and Western blotting in ovarian cancer samples. On the basis of these results, Id-1 protein expression was determined by immunohistochemistry in 101 specimens of epithelial ovarian cancer, in 40 borderline tumors, and in 20 cystadenomas. In these cases, Id-1 expression was correlated with p21 expression, microvessel density, and survival. RESULTS: By immunohistochemistry, detectable expression of Id-1 was found significantly more often in ovarian cancers (74.3%) than in borderline tumors (32.5%) and cystadenomas (0%; P < 0.0001, chi(2) test). Cancer samples with poor or moderate histological differentiation (G3/G2) showed significantly stronger Id-1 expression than cancer samples with high differentiation (G1; P = 0.021, Mann Whitney test), and no association of Id-1 with p21 expression or microvessel density was found. In cancer samples strong or moderate expression of Id-1 was a strong predictor for shorter overall survival in uni- and multivariate analysis (P = 0.001, Cox-regression). CONCLUSIONS: The level of Id-1 protein expression correlates with the malignant potential of ovarian tumors. In cancer samples, stronger Id-1 expression is associated with poor differentiation and more aggressive behavior of tumor cells, resulting in poor clinical outcome. Consequently, Id-1 inhibition in the future might be of benefit for patients with ovarian cancer. PMID- 12576451 TI - Combination of dihydropyrimidine dehydrogenase and thymidylate synthase gene expressions in primary tumors as predictive parameters for the efficacy of fluoropyrimidine-based chemotherapy for metastatic colorectal cancer. AB - Dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) gene expressions in metastatic colorectal cancer have been reported to be predictive parameters for the efficacy of fluoropyrimidine-based chemotherapy. In this study, we investigated the association between both DPD and TS expressions in primary colorectal tumor and the antitumor effect in patients with metastatic colorectal cancer when treated with a fluoropyrimidine-based protocol. DPD and TS expressions were measured by reverse transcription-PCR in surgically resected materials of primary colorectal tumors from 37 patients who went on to receive oral treatment of uracil and tegafur and leucovorin for either synchronous or metachronous metastatic diseases. Relative mRNA amounts of DPD or TS were expressed as the ratios of targeted gene to glyceraldehyde-3-phosphate dehydrogenase reverse transcription-PCR products. Median values of DPD mRNA expressions were 0.30 and 0.65 for responding tumors and nonresponding ones, respectively, with a statistical significance (P < 0.0001). No responding tumor had a DPD mRNA expression >/= 0.5. A total of 19 tumors had low DPD mRNA expressions of <0.5, and 63% of them showed response. There was no responding tumor with both high DPD and high TS (TS mRNA expression >/= 1.0). However, the response rate was 75% in tumors with both low DPD and low TS. The median survival time was 16.3 months in patients with both low DPD and low TS versus 8.4 months in patients with high DPD or high TS mRNA expression. In conclusion, the combination of DPD and TS mRNA expressions in the primary tumor might be useful as predictive parameters for the efficacy of fluoropyrimidine-based chemotherapy for metastatic colorectal cancer. PMID- 12576452 TI - Thymidylate synthase expression in advanced colorectal cancer predicts for response to raltitrexed. AB - PURPOSE: The purpose of this research was to evaluate the predictive value of expression of thymidylate synthase (TS) and other genes for response to raltitrexed (RTX). EXPERIMENTAL DESIGN: Twenty-five patients with metastatic colorectal cancer received RTX 3 mg/m(2) 3-weekly. Pretreatment tumor biopsies were analyzed for TS, dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP), folylpolyglutamate synthetase, and reduced folate carrier mRNA expression by real-time reverse transcription-PCR. TS protein expression was evaluated by immunohistochemistry using a polyclonal TS antibody. RESULTS: Twenty patients were evaluable for response and gene expression. Six of 20 (30%) achieved a partial response. Median TS/beta-actin was 5.7 x 10(3) (range, 2.2-42 x 10(3)). Median TS/beta-actin was 3.7 x 10(3) in responding patients and 6.1 x 10(3) in nonresponders (P = 0.048). Five of 6 patients with TS/beta-actin 5% p27(Kip1) positive myeloma cells). RESULTS: Low p27(Kip1) expression was observed in 23 (31%) patients. The response rate to standard dose chemotherapy (including vincristine, doxorubicin, and dexamethasone induction before high-dose chemotherapy) was 70%, with no significant difference between patients with low or high p27(Kip1) expression (83 versus 65%; P = 0.1). Kaplan-Meier analysis of all 74 patients revealed that patients with low p27(Kip1) expression had a significantly shorter overall survival (median, 3.7 years versus 4.7 years; P = 0.03) than those with high p27(Kip1) expression. Patients with high p27(Kip1) expression receiving high-dose chemotherapy experienced prolonged overall survival as compared with those with low p27(Kip1) expression (median not yet reached versus 2.9 years; P = 0.008). By multivariate Cox regression analyses, low p27(Kip1)(P = 0.03), deletion of chromosome 13q14 (P = 0.02), and beta(2) microglobulin (P = 0.01) were identified as independent adverse prognostic factors for overall survival. According to the number of independent unfavorable prognostic factors present in each patient, low-risk, intermediate-risk, and high risk patients with different overall survival times were defined (median overall survival, 6.3 versus 4.2 versus 1.8 years; P < 0.001). CONCLUSIONS: Low p27(Kip1) expression is an independent adverse prognostic factor in patients with MM. The proposed risk score might be useful for risk-adapted treatment in the future. PMID- 12576456 TI - RNA expression of breast cancer resistance protein, lung resistance-related protein, multidrug resistance-associated proteins 1 and 2, and multidrug resistance gene 1 in breast cancer: correlation with chemotherapeutic response. AB - PURPOSE: The aim of this study was to investigate whether expression of particular drug resistance genes in primary operable breast cancer correlates with response to first-line chemotherapy in advanced disease. EXPERIMENTAL DESIGN: We determined mRNA levels of BCRP, LRP, MRP1, MRP2, and MDR1 in 59 primary breast tumor specimens of patients who received chemotherapy as first line systemic treatment after diagnosis of advanced disease. The relative expression levels were measured by quantitative real-time reverse transcription PCR and subsequently analyzed in relation to the type of response to chemotherapy, the length of progression-free survival (PFS), and post-relapse overall survival. RESULTS: For each of these drug resistance genes, a large variation in expression level was observed among the tumors of the different patients. When analyzing mRNA expression in relation to overall response, it was found that the median expression level of these five drug resistance genes in the responding tumors, as compared with nonresponding tumors, was markedly lower. Classification of tumors as high versus low with respect to the expression level of these genes showed that the overall response in the MDR1-high subset (17%), as compared with the MDR1-low subset (68%), was significantly lower (P = 0.005). Although similar differences in response rate were found for subsets of tumors stratified by the expression level of the other drug resistance genes, none of the observed differences were statistically significant. However, in the subgroup of patients treated with anthracycline-based chemotherapy (5-fluorouracil, Adriamycin/epirubicin, and cyclophosphamide), a correlation between response and the expression of BCRP and MRP1 (only PFS) was found, whereas such an association was not present in the cyclophosphamide, methotrexate, and 5-fluorouracil-treated group of patients. Furthermore, high expression of LRP as well as MDR1 was found to be significantly associated with a poor PFS (P = 0.04 and P < 0.001, respectively). For lung resistance-related protein, this association was limited to 5-fluorouracil, Adriamycin/epirubicin, and cyclophosphamide. Expression levels of BCRP, MRP1, or MRP2 were not related with the length of PFS. Furthermore, no correlation between the expression level of these drug resistance genes and post relapse overall survival was found. CONCLUSIONS: In this pilot study, MDR1 expression in primary breast tumors was inversely related with the efficacy of first-line chemotherapy, and high expression level was a significant predictor of poor prognosis for patients with advanced disease. Apart from MDR1, the expression levels of BCRP, LRP, and MRP1 might have some additional predictive value for clinical outcome. PMID- 12576457 TI - Sequence alterations in the reduced folate carrier are observed in osteosarcoma tumor samples. AB - High-dose methotrexate is a standard component of therapy for high-grade osteosarcoma. Its effectiveness may be limited by intrinsic and acquired resistance. Decreased reduced folate carrier (RFC) expression has been shown in approximately half of osteosarcomas at diagnosis. Mutations and polymorphisms in the RFC gene have been reported in various cell lines. The purpose of this study was to investigate sequence alterations in the RFC gene in osteosarcoma tumor samples. The entire coding region of the RFC gene in samples from 162 osteosarcoma patients was screened by DNA single-stranded conformational polymorphism, followed by direct sequencing of any region with altered mobility. A previously identified polymorphism at cDNA position number 174 of RFC exon 2 was observed. Sixty-one samples (37.6%) were heterozygous with both A/G at this position (His(27)/Arg(27)), 52 samples (32.2%) were homozygous with G (Arg(27)), and 49 samples (30.2%) were homozygous with A (His(27)). Fifteen (9.2%) samples were identified with other RFC sequence variants in exon 2, none of which have been reported. The sequence variants in exon 2 included a G to A substitution at cDNA position 231, a G to A substitution at cDNA position 155, a C to T substitution at cDNA position 114, and a T to C substitution at cDNA position 104, resulting in a serine to asparagine substitution at amino acid 46, a glutamate to lysine substitution at amino acid 21, an alanine to valine substitution at amino acid 7, and a serine to proline substitution at amino acid 4, respectively. A deletion of A at cDNA position 126 resulting in a frameshift was also observed. Some of these variants were observed in multiple samples. Eight samples had altered single-stranded conformational polymorphism patterns in exon 3 that were associated with nucleotide changes that altered the amino acid sequence. All of these RFC sequence variants appeared to be heterozygous. Heterozygous C/T and homozygous C also were observed at RFC cDNA position 790 in exon 3, which does not alter the amino acid coding sequence. This study shows that RFC sequence alterations are frequent in samples from osteosarcoma patients. Additional studies are under way to determine the clinical significance of these sequence alterations and their effect on methotrexate transport and resistance. PMID- 12576458 TI - Establishment and characterization of cancer cell cultures and xenografts derived from primary or metastatic Mullerian cancers. AB - PURPOSE: The purpose of this study was to characterize cell cultures and xenografts derived from patients with ovarian cancer. EXPERIMENTAL DESIGN: Ninety specimens from 67 patients were plated in RPMI 1640 or inoculated in nude mice. Growth characteristics of cell cultures and xenografts were determined. Expression of receptors for estrogen, progesterone, androgen, epithelial growth factor, fibroblast growth factor, HER-2/erbB-2/c-neu proto-oncogene, and the P53 expression were characterized by immunocytochemistry in 28 cell cultures. RESULTS: Forty-nine percent of samples were cultured successfully in vitro. Ascitic and pleural effusion specimens were more likely to produce a cell culture or a xenograft than solid tissue specimens (P < 0.005). All of the cell cultures had an epithelial morphology, and 89% were aneuploid with a mean DNA index of 1.6 (range, 0.9-3.0). Of 54 and 61 specimens inoculated into nude mice i.p. and s.c., 15 (28%) and 18 (30%) produced a xenograft, respectively, with two-thirds of these xenografts being reproducibly tumorigenic. The median time to first passage was 21 weeks for cell cultures and 8-12 weeks for xenografts. Expression of epithelial growth factor receptor, HER-2/erbB-2/c-neu proto-oncogene, fibroblast growth factor receptor, estrogen, progesterone, and androgen was seen in 24, 21, 31, 17, 43, and 18%, respectively. P53 was overexpressed in 62% of cell cultures analyzed. CONCLUSIONS: Ovarian cancer cells collected from effusions are easier to grow in vitro than in vivo. The only characteristic that may be associated with tumorigenicity was abnormal P53 expression. This panel of ovarian cancer materials provides useful models for biological or therapeutical studies. PMID- 12576459 TI - Macrophage migration inhibitory factor and CXC chemokine expression in non-small cell lung cancer: role in angiogenesis and prognosis. AB - PURPOSE: The purpose of this study was to determine whether expression of migration inhibitory factor (MIF) is increased in non-small cell lung cancer, and whether it correlates with angiogenesis and/or prognosis. EXPERIMENTAL DESIGN: We measured vessel density, and levels of MIF, angiogenic CXC chemokines, and vascular-endothelial growth factor (VEGF; by ELISA) in tumor and normal lung tissue from 87 patients after resection of lung cancer. We compared vessel density with levels of MIF, VEGF, or angiogenic CXC chemokines in the corresponding tumor homogenate. Disease-free survival was analyzed in a Cox proportional hazards model. RESULTS: Levels of MIF in lung cancer demonstrated a bimodal distribution, with some having "normal" values (relative to normal lung tissue) and a second cluster with markedly high values. The increased levels of MIF in lung cancer were statistically significant in both paired and unpaired comparisons (P < 0.05). The strongest correlation of vessel density was with the sum of angiogenic CXC chemokines. MIF correlated very strongly with levels of angiogenic CXC chemokines. Tumors in the high MIF group had a strong correlation between MIF level and vessel density. Risk of recurrence was associated with high levels of glutamic acid-leucine-arginine amino acid motif CXC chemokines, MIF, and/or VEGF in a Cox proportional hazards model. CONCLUSIONS: MIF expression in non-small cell lung cancer occurs in a bimodal distribution, and is closely associated with tumor levels of angiogenic CXC chemokines and with vessel density. High levels of tumor-associated CXC chemokines, MIF, or VEGF are associated with risk of recurrence after resection of lung cancer. PMID- 12576460 TI - Apoptosis by leukemia cell-targeted diphtheria toxin occurs via receptor independent activation of Fas-associated death domain protein. AB - PURPOSE: We examined the mechanism of action of a targeted fusion toxin consisting of diphtheria toxin fused to granulocyte macrophage colony stimulating factor (GMCSF) (DT(388)-GMCSF), which was designed to selectively kill acute myeloid leukemia cells. EXPERIMENTAL DESIGN AND RESULTS: U937 cells treated with DT(388)-GMCSF underwent apoptosis as shown by chromatin degradation and cellular and nuclear fragmentation. This apoptosis was prevented by a general caspase inhibitor. DT(388)-GMCSF treatment resulted in activation of the initiator caspases 8 and 9 and effector caspases. A selective caspase 8 inhibitor prevented activation of caspase 9, whereas a selective caspase 9 inhibitor did not prevent activation of caspase 8, indicating that caspase 8 activation is the proximal event in DT(388)-GMCSF-induced apoptosis. Caspase 8 was activated through a Fas associated death domain protein (FADD)-dependent mechanism as demonstrated by inhibition of DT(388)-GMCSF-induced apoptosis on expression of a dominant negative FADD molecule. However, unlike most FADD-dependent apoptosis, this pathway may not involve death receptors, including Fas, tumor necrosis factor receptor 1, or tumor necrosis factor-related apoptosis-inducing ligand receptors, because inhibitors of the receptors did not prevent DT(388)-GMCSF-induced apoptosis. CONCLUSIONS: These data indicate that targeted toxins induce apoptosis by activating components of the death receptor pathway in a receptor-independent manner. PMID- 12576461 TI - Combined targeted inhibition of bcl-2, bcl-XL, epidermal growth factor receptor, and protein kinase A type I causes potent antitumor, apoptotic, and antiangiogenic activity. AB - PURPOSE: This study investigated whether the functional and structural interactions between epidermal growth factor receptor (EGFR), protein kinase AI (PKAI), and bcl-2/bcl-xL could be exploited to obtain cooperative antitumor effects against models of human colon and breast cancer. EXPERIMENTAL DESIGN: Antisense bcl-2/bcl-xL (4625), antisense PKAI (AS-PKAI), and ZD1839 ("Iressa"), a selective EGFR tyrosine kinase inhibitor, were administered as single agents and in combination against GEO colon and ZR-75-1 breast cancer cell lines in vitro and to mice bearing s.c. GEO human tumor xenografts in vivo. Effects on growth inhibition, vascular endothelial growth factor secretion, and induction of apoptosis were assessed. RESULTS: Antisense bcl-2/bcl-xL inhibited the growth of GEO and ZR-75-1 cells in vitro, reducing bcl-2 and bcl-xL expression and vascular endothelial growth factor secretion. Supra-additive growth inhibition and apoptosis induction were observed when 4625 was combined with ZD1839 or AS-PKAI. Combining all three agents resulted in a complete growth inhibitory effect in vitro. Antisense bcl-2/bcl-xL, AS-PKAI, and ZD1839 administered in vivo as single agents caused growth inhibition of GEO xenografts. Combining all three agents caused a marked and sustained effect, with 50% growth inhibition and 50% of mice tumor free 5 weeks after treatment withdrawal. The combination was well tolerated. CONCLUSIONS: The combination of 4625, AS-PKAI, and ZD1839 resulted in a strong antiproliferative, proapoptotic, and antiangiogenic response, suggestive of a functional interaction between EGFR, PKAI, and bcl-2/bcl-xL and providing a rationale for the selection of specific molecular treatments for the development of therapeutic strategies. Iressa is a trademark of the AstraZeneca group of companies. PMID- 12576462 TI - Microglia cyclooxygenase-2 activity in experimental gliomas: possible role in cerebral edema formation. AB - PURPOSE: Cerebral edema is responsible for significant morbidity and mortality in patients harboring malignant gliomas. To examine the role of inflammatory cells in brain edema formation, we studied the expression cyclooxygenase (COX)-2, a key enzyme in arachidonic acid metabolism, by microglia in the C6 rodent glioma model. EXPERIMENTAL DESIGN: The expression of COX-2 in primary microglia cultures obtained from intracranial rat C6 gliomas was examined using reverse transcription-PCR, Western analysis, and prostaglandin E(2) (PGE(2)) enzyme immunoassay. Blood-tumor barrier permeability was studied in the same tumor model using magnetic resonance imaging. RESULTS: In contrast to C6 glioma cells, microglia isolated from intracranial C6 tumors produced high levels of PGE(2) through a COX-2-dependent pathway. To test whether the observed microglia COX-2 activity played a role in brain edema formation in gliomas, tumor-bearing rats were treated with rofecoxib, a selective COX-2 inhibitor. Rofecoxib was as effective as dexamethasone in decreasing the diffusion of contrast material into the brain parenchyma (P = 0.01, rofecoxib versus control animals), suggesting a reduction in blood-tumor barrier permeability. CONCLUSIONS: These findings suggest that glioma-infiltrating microglia are a major source of PGE(2) production through the COX-2 pathway and support the use of COX-2 inhibitors as possible alternatives to glucocorticoids in the treatment of peritumoral edema in patients with malignant brain tumors. PMID- 12576463 TI - Phytosphingosine induces apoptotic cell death via caspase 8 activation and Bax translocation in human cancer cells. AB - PURPOSE: Sphingolipid metabolites, such as sphingosine and ceramide, are highly bioactive compounds and are involved in diverse cell processes, including cell cell interaction, cell proliferation, differentiation, and apoptosis. However, the physiological roles of phytosphingosine are poorly understood. In this study, we report that phytosphingosine can potently induce apoptotic cell death in human cancer cells via caspase activation and caspase-independent cytochrome c release. EXPERIMENTAL DESIGN: Phytosphingosine-induced apoptosis was determined by Hoechst 33258 staining, flow cytometric analysis, and DNA fragmentation assay. Involvement of caspases was determined by immunoblot analysis and cell death detection assays after treatment with synthetic inhibitor z-Val-Ala-Asp fluoromethyl ketone, z-DEVD-fmk, or z-IETD-fmk. Death receptor (DR) dependency was analyzed by examining expression of DRs (Fas, DR4, DR5, TNFR1, and R2), and interaction of Fas-associated death domain and caspase 8. Involvement of the mitochondria pathway was examined by monitoring of the mitochondria membrane potential, cytochrome c release, and Bax translocation. RESULTS: Phytosphingosine treated cells displayed several features of apoptosis, including increase of sub G(1) population, DNA fragmentation, and poly(ADP-ribose) polymerase cleavage. We observed that phytosphingosine cause activation of caspase 8 in a DR-independent fashion. Phytosphingosine also induced activation of caspase 9 and 3, loss of mitochondrial membrane potential, and the cytochrome c release from mitochondria. However, we failed to detect Bid cleavage. Moreover, caspase 8 inhibitor z-IETD fmk did not affect phytosphingosine-induced cytochrome c release and caspase 9 activation, suggesting that phytosphingosine-induced cytochrome c release is caused by caspase 8-independent manner. Phytosphingosine induced mitochondrial translocation of Bax from the cytosol without changes in the protein levels of Bcl-2, Bcl-xL, and Bax. In addition, Bcl-2/Bax interaction was diminished after addition of phytosphingosine. CONCLUSION: These findings indicate that phytosphingosine induces apoptotic cell death in human cancer cells by direct activation of caspase 8, and by mitochondrial translocation of Bax and subsequent release of cytochrome c into cytoplasm, providing a potential mechanism for the anticancer activity of phytosphingosine. PMID- 12576464 TI - Docetaxel enhances the therapeutic effect of the angiogenesis inhibitor TNP-470 (AGM-1470) in metastatic human transitional cell carcinoma. AB - We demonstrated recently that chronic frequent administration of an adequate biological dose of the angiogenesis inhibitor TNP-470 (AGM-1470, O-chloracetyl carbamoyl fumagillol) completely inhibits spontaneous lymph node metastasis but does not have a complete response on tumor growth of nonestablished or established human metastatic transitional cell carcinoma (TCC) 253J B-V growing orthotopically into athymic nude mice. Therefore, in this study, we evaluated whether docetaxel (Taxotere) enhances the therapeutic effect of TNP-470, especially on tumor growth. Docetaxel enhanced in vitro antiproliferation but not basic fibroblast growth factor down-regulation by TNP-470 in 253J B-V and human umbilical vascular endothelial cells. Docetaxel significantly enhanced in vitro apoptosis by TNP-470 in human umbilical vascular endothelial cells but not in 253J B-V. In vivo combination was most effective when docetaxel was administered before TNP-470, and increased significantly the complete response on tumor growth of nonestablished and established TCCs growing orthotopically into athymic nude mice compared with either therapy alone (P < 0.05). The incidence of spontaneous lymph node metastasis was inhibited completely by the combination therapy (P < 0.05). Drug-induced body weight loss was not significantly different in any treatment groups. The combination of TNP-470 and docetaxel inhibited intratumor neovascularization, the expression of bFGF and matrix metalloproteinases type-9 compared with controls (P < 0.005), and enhanced apoptosis in tumors compared with each therapy alone (P < 0.005). These studies indicate that docetaxel markedly enhances the ability of TNP-470 to inhibit tumorigenicity and metastasis in both nonestablished and established TCCs. These effects are mediated, in part, by the complementary cytotoxicities of angiogenesis inhibition, down-regulation of bFGF and matrix metalloproteinases type-9, and induction of apoptosis. PMID- 12576465 TI - In vitro schedule-dependent interaction between docetaxel and gemcitabine in human gastric cancer cell lines. AB - PURPOSE: The purpose of this study was to assess the activity of clinically used drugs and to define the most effective treatment scheme in human gastric cancer cell lines. EXPERIMENTAL DESIGN: Cytotoxic activity was evaluated by sulforhodamine B assay, potential clinical activity was estimated by relative antitumor activity, and the type of drug interaction was assessed using the method of Chou and Talalay. Cell cycle perturbations and apoptosis were evaluated by flow cytometry, mitotic index by microscopic analysis, bax, bcl-2, and p53 by immunohistochemistry, and cyclin B expression by Western blot. RESULTS: Gemcitabine (GEM) and docetaxel (DOC) were the most potent of the seven drugs tested, with maximum relative antitumor activity values in all of the cell lines. Simultaneous treatment with GEM and DOC, and the sequence GEM-->DOC caused an antagonistic interaction, as shown by the combination index >1, at all levels of killed cell fraction. Conversely, the sequential treatment DOC-->GEM produced a synergistic interaction (combination index < 1). On the basis of cell cycle perturbations, it can be hypothesized that the antimetabolite (GEM) attacks cells recovering rapidly from an M block induced by DOC as they progress to the S phase, producing a powerful cytocidal effect, as shown by the increase from 15 to 75% of apoptotic cells. CONCLUSIONS: Our findings suggest that the interaction of DOC and GEM is highly schedule dependent and has been used recently to plan a Phase I-II clinical protocol. PMID- 12576466 TI - Clinical evaluation of mitoxantrone and piroxicam in a canine model of human invasive urinary bladder carcinoma. AB - PURPOSE: Cyclooxygenase inhibitors show promise in chemoprevention and therapy of certain carcinomas, an effect that may be additive to that of standard chemotherapy. The purpose of this study was to evaluate the efficacy of combined therapy using the cyclooxygenase inhibitor, piroxicam, and mitoxantrone against a relevant canine model of human invasive bladder cancer. EXPERIMENTAL DESIGN: Fifty-five dogs with transitional cell carcinoma of the urinary bladder were enrolled in this nonrandomized one-armed prospective multi-institutional clinical trial. Mitoxantrone was administered i.v. (5 mg/m(2)) every 21 days for four treatments, and piroxicam was administered p.o. (0.3 mg/kg/day) for the study duration. Tumor staging was performed at baseline, day 42 and every 3 months after protocol completion. Endpoints included time-to-treatment failure and survival time (ST). RESULTS: Response data were available for 48 dogs and included one complete response, 16 partial responses, 22 with disease stabilization, and 9 with progressive disease for an overall 35.4% measurable response rate. Subjective improvement occurred in 75% of treated dogs. Median time-to-treatment failure and ST were 194 and 350 days, respectively. Using censoring and end point definitions similar to those of previous reports of dogs treated with piroxicam alone, the median ST in this study was 291 days, compared with 181 days with piroxicam alone. Diarrhea and azotemia were the most common treatment complications. CONCLUSIONS: Mitoxantrone/piroxicam induced remission more frequently than previously reported for either drug as a single agent in this canine model of invasive human transitional cell carcinoma. Additional evaluation of these drugs in combination protocols should be explored. PMID- 12576467 TI - Correspondence re: C. G. Ferreira et al., Apoptosis: target of cancer therapy. Clin. Cancer Res., 8: 2024-2034, 2002. PMID- 12576469 TI - Polysialic acid in human milk. CD36 is a new member of mammalian polysialic acid containing glycoprotein. AB - The neural cell adhesion molecule and the voltage-sensitive sodium channel alpha subunit are the only two molecules in mammals known to be modified by alpha-2,8 linked polysialic acid (polySia). We found a new polySia-containing glycoprotein in human milk and identified it as CD36, a member of the B class of the scavenger receptor superfamily. The polySia-containing glycan chain(s) were removed by alkaline treatment but not by peptide:N-glycanase F digestion, indicating that milk CD36 contained polySia on O-linked glycan chain(s). Polysialylation of CD36 occurs not only in human milk but also in mouse milk. However, CD36 in human platelets is not polysialylated. PolySia CD36 is secreted in milk at any lactation stage and reaches peak level at 1 month after parturition. Thus, it is suggested that polySia of milk CD36 is significant for neonatal development in terms of protection and nutrition. PMID- 12576470 TI - Asymmetric organization of the pore region of the epithelial sodium channel. AB - Epithelial sodium channels (ENaCs) are composed of three homologous subunits that have regions preceding the second transmembrane domain (also referred as pre-M2) that form part of the channel pore. To identify residues within this region of the beta-subunit that line the pore, we systematically mutated residues Gln(523) Ile(536) to cysteine. Wild type and mutant mouse ENaCs were expressed in Xenopus oocytes, and a two-electrode voltage clamp was used to examine the properties of mutant channels. Cysteine substitutions of 9 of 13 residues significantly altered Li(+) to Na(+) current ratios, whereas only cysteine replacement of beta Gly(529) resulted in K(+)-permeable channels. Besides beta G525C, large increases in the inhibitory constant of amiloride were observed with mutations at beta Gly(529) and beta Ser(531) within the previously identified 3-residue tract that restricts K(+) permeation. Cysteine substitution preceding (beta Phe(524) and beta Gly(525)), within (beta Gly(530)) or following (beta Leu(533)) this 3-residue tract, resulted in enhanced current inhibition by external MTSEA. External MTSET partially blocked channels with cysteine substitutions at beta Gln(523), beta Phe(524), and beta Trp(527). MTSET did not inhibit alpha beta G525C gamma, although previous studies showed that channels with cysteine substitutions at the corresponding sites within the alpha- and gamma-subunits were blocked by MTSET. Our results, placed in context with previous observations, suggest that pore regions from the three ENaC subunits have an asymmetric organization. PMID- 12576471 TI - Three-dimensional localization of divergent region 3 of the ryanodine receptor to the clamp-shaped structures adjacent to the FKBP binding sites. AB - Of the three divergent regions of ryanodine receptors (RyRs), divergent region 3 (DR3) is the best studied and is believed to be involved in excitation contraction coupling as well as in channel regulation by Ca(2+) and Mg(2+). To gain insight into the structural basis of DR3 function, we have determined the location of DR3 in the three-dimensional structure of RyR2. We inserted green fluorescent protein (GFP) into the middle of the DR3 region after Thr-1874 in the sequence. HEK293 cells expressing this GFP-RyR2 fusion protein, RyR2(T1874-GFP,) were readily detected by their green fluorescence, indicating proper folding of the inserted GFP. RyR2(T1874-GFP) was further characterized functionally by assays of Ca(2+) release and [(3)H]ryanodine binding. These analyses revealed that RyR2(T1874-GFP) functions as a caffeine- and ryanodine-sensitive Ca(2+) release channel and displays Ca(2+) dependence and [(3)H]ryanodine binding properties similar to those of the wild type RyR2. RyR2(T1874-GFP) was purified from cell lysates in a single step by affinity chromatography using GST-FKBP12.6 as the affinity ligand. The three-dimensional structure of the purified RyR2(T1874-GFP) was then reconstructed using cryoelectron microscopy and single particle image analysis. Comparison of the three-dimensional reconstructions of wild type RyR2 and RyR2(T1874-GFP) revealed the location of the inserted GFP, and hence the DR3 region, in one of the characteristic domains of RyR, domain 9, in the clamp-shaped structure adjacent to the FKBP12 and FKBP12.6 binding sites. COOH-terminal truncation analysis demonstrated that a region between 1815 and 1855 near DR3 is essential for GST-FKBP12.6 binding. These results provide a structural basis for the role of the DR3 region in excitation-contraction coupling and in channel regulation. PMID- 12576472 TI - Activation of the RegB endoribonuclease by the S1 ribosomal protein is due to cooperation between the S1 four C-terminal modules in a substrate-dependant manner. AB - The RegB protein, encoded by the T4 bacteriophage genome, is a ribonuclease involved in the inactivation of the phage early messenger RNAs. Its in vitro activity is very low but can be enhanced up to 100-fold in the presence of the ribosomal protein S1. The latter is made of six repeats of a conserved module found in many other proteins of RNA metabolism. Considering the difference between its size (556 amino acids) and that of several RegB substrates (10 nucleotides), we wondered whether all six modules are necessary for RegB activation. We studied the influence of twelve S1 fragments on the cleavage efficiency of three short substrates. RegB activation requires the cooperation of different sets of modules depending on the substrates. Two RNAs are quite well cleaved in the presence of the fragment formed by the fourth and fifth modules, whereas the third requires the presence of the four C-terminal domains. However, NMR interaction experiments showed that, despite these differences, the interactions of the substrates with either the bi- or tetra-modules are similar, suggesting a common interaction surface. In the case of the tetra-module the interactions involve all four domains, raising the question of the spatial organization of this region. PMID- 12576473 TI - Comprehensive analysis of the secreted proteins of the parasite Haemonchus contortus reveals extensive sequence variation and differential immune recognition. AB - Haemonchus contortus is a nematode that infects small ruminants. It releases a variety of molecules, designated excretory/secretory products (ESP), into the host. Although the composition of ESP is largely unknown, it is a source of potential vaccine components because ESP are able to induce up to 90% protection in sheep. We used proteomic tools to analyze ESP proteins and determined the recognition of these individual proteins by hyperimmune sera. Following two dimensional electrophoresis of ESP, matrix-assisted laser desorption ionization time-of-flight and liquid chromatography-tandem mass spectrometry were used for protein identification. Few sequences of H. contortus have been determined. Therefore, the data base of expressed sequence tags (dbEST) and a data base consisting of contigs from Haemonchus ESTs were also consulted for identification. Approximately 200 individual spots were observed in the two dimensional gel. Comprehensive proteomics analysis, combined with bioinformatic search tools, identified 107 proteins in 102 spots. The data include known as well as novel proteins such as serine, metallo- and aspartyl proteases, in addition to H. contortus ESP components like Hc24, Hc40, Hc15, and apical gut GA1 proteins. Novel proteins were identified from matches with H. contortus ESTs displaying high similarity with proteins like cyclophilins, nucleoside diphosphate kinase, OV39 antigen, and undescribed homologues of Caenorhabditis elegans. Of special note is the finding of microsomal peptidase H11, a vaccine candidate previously regarded as a "hidden antigen" because it was not found in ESP. Extensive sequence variation is present in the abundant Hc15 proteins. The Hc15 isoforms are differentially recognized by hyperimmune sera, pointing to a possible specific role of Hc15 in the infectious process and/or in immune evasion. This concept and the identification of multiple novel immune-recognized components in ESP should assist future vaccine development strategies. PMID- 12576474 TI - Smad4 as a transcription corepressor for estrogen receptor alpha. AB - Antiestrogen compounds exhibit a variety of different effects in different tissues and are widely used for the treatment of osteoporosis, breast cancer, and other diseases. Upon examining the molecular mechanisms, we found that Smad4, a common signal transducer in the bone morphogenetic protein (BMP)/transforming growth factor-beta (TGF-beta) signaling pathway, functions as a transcription corepressor for human estrogen receptor alpha (ERalpha). Endogenous ERalpha was co-immunoprecipitated with Smad4, and the interaction was induced by antiestrogen ligands such as tamoxifen, raloxifene, and droloxifen, which was confirmed in chromatin immunoprecipitation assays. Smad4 and ERalpha form a complex when ERalpha binds to the estrogen-responsive element within the estrogen target gene promoter. Importantly, the expression of Smad4 inhibits both antiestrogen-induced luciferase activity and estrogen downstream target gene transcription in breast cancer cells. Mapping of the interaction domains indicates that the activation function 1 (AF1) domain of ERalpha is essential for its interaction with Smad4, while the MH1 domain and linker region of Smad4 are essential for the interaction. Our findings represent a novel mechanism that TGF-beta may regulate cell fate through Smad4-mediated cross-talk with estrogen. PMID- 12576475 TI - mRNA capping enzyme requirement for Caenorhabditis elegans viability. AB - Capping of the initiated 5' ends of RNA polymerase II products is evolutionarily and functionally conserved from yeasts to humans. The m(7)GpppN cap promotes RNA stability, processing, transport, and translation. Deletion of capping enzymes in yeasts was shown to be lethal due to rapid exonucleolytic degradation of uncapped transcripts or failure of capped but unmethylated RNA to initiate protein synthesis. Using RNA interference and Caenorhabditis elegans we have found that RNA capping is also essential for metazoan viability. C. elegans bifunctional capping enzyme was cloned, and capping activity by the expressed protein as well as growth complementation of yeast deletion strains missing either RNA triphosphatase or guanylyltransferase required terminal sequences not present in the previously isolated cel-1 clone. By RNA interference analysis we show that cel-1 is required for embryogenesis. cel-1(RNAi) embryos formed cytoplasmic granules characteristic of a phenocluster of RNA processing genes and died early in development. PMID- 12576477 TI - Examining the role of glutamic acid 183 in chloroperoxidase catalysis. AB - Site-directed mutagenesis has been used to investigate the role of glutamic acid 183 in chloroperoxidase catalysis. Based on the x-ray crystallographic structure of chloroperoxidase, Glu-183 is postulated to function on distal side of the heme prosthetic group as an acid-base catalyst in facilitating the reaction between the peroxidase and hydrogen peroxide with the formation of Compound I. In contrast, the other members of the heme peroxidase family use a histidine residue in this role. Plasmids have now been constructed in which the codon for Glu-183 is replaced with a histidine codon. The mutant recombinant gene has been expressed in Aspergillus niger. An analysis of the produced mutant gene shows that the substitution of Glu-183 with a His residue is detrimental to the chlorination and dismutation activity of chloroperoxidase. The activity is reduced by 85 and 50% of wild type activity, respectively. However, quite unexpectedly, the epoxidation activity of the mutant enzyme is significantly enhanced approximately 2.5-fold. These results show that Glu-183 is important but not essential for the chlorination activity of chloroperoxidase. It is possible that the increased epoxidation of the mutant enzyme is based on an increase in the hydrophobicity of the active site. PMID- 12576476 TI - The Caenorhabditis elegans mRNA 5'-capping enzyme. In vitro and in vivo characterization. AB - Eukaryotic mRNA capping enzymes are bifunctional, carrying both RNA triphosphatase (RTPase) and guanylyltransferase (GTase) activities. The Caenorhabditis elegans CEL-1 capping enzyme consists of an N-terminal region with RTPase activity and a C-terminal region that resembles known GTases, However, CEL 1 has not previously been shown to have GTase activity. Cloning of the cel-1 cDNA shows that the full-length protein has 623 amino acids, including an additional 38 residues at the C termini and 12 residues at the N termini not originally predicted from the genomic sequence. Full-length CEL-1 has RTPase and GTase activities, and the cDNA can functionally replace the capping enzyme genes in Saccharomyces cerevisiae. The CEL-1 RTPase domain is related by sequence to protein-tyrosine phosphatases; therefore, mutagenesis of residues predicted to be important for RTPase activity was carried out. CEL-1 uses a mechanism similar to protein-tyrosine phosphatases, except that there was not an absolute requirement for a conserved acidic residue that acts as a proton donor. CEL-1 shows a strong preference for RNA substrates of at least three nucleotides in length. RNA mediated interference in C. elegans embryos shows that lack of CEL-1 causes development to arrest with a phenotype similar to that seen when RNA polymerase II elongation activity is disrupted. Therefore, capping is essential for gene expression in metazoans. PMID- 12576478 TI - Soluble organic matrices of the calcitic prismatic shell layers of two Pteriomorphid bivalves. Pinna nobilis and Pinctada margaritifera. AB - The calcitic prisms of the shells of two bivalves, Pinna and Pinctada, are considered simple prisms according to some morphological and mineralogical characteristics. Scanning electron microscopic and atomic force microscopic studies show that the microstructures and nanostructures of these two shells are different. Pinna prisms are monocrystalline, whereas Pinctada prisms are not. Moreover, intraprismatic membranes are present only in the Pinctada prisms. The soluble organic matrices extracted from these prisms are acidic, but their bulk compositions differ. Ultraviolet and infrared spectrometries, fluorescence, high pressure liquid chromatography, and electrophoresis show that the sugar-protein ratios and the molecular weights are different. Sulfur is mainly associated with acidic sulfated sugars, not with amino acids, and the role of acidic sulfated sugars is still underestimated. Thus, the simple prism concept is not a relevant model for the biomineralization processes in the calcitic prismatic layer of mollusk shells. PMID- 12576479 TI - Cloning and functional characterization of a mouse intestinal acyl CoA:monoacylglycerol acyltransferase, MGAT2. AB - Acyl-CoA:monoacylglycerol transferase (MGAT) plays a predominant role in dietary fat absorption in the small intestine, where it catalyzes the first step of triacylglycerol resynthesis in enterocytes for chylomicron formation and secretion. Although the mouse small intestine exhibits the highest MGAT enzyme activity among all of the tissues studied, the gene encoding the enzyme has not been identified so far. In the present studies, we report the identification and characterization of a mouse intestinal MGAT, MGAT2. Transient expression of MGAT2 in AV-12, COS-7, and Caco-2 cells led to a more than 70-, 30-, and 35-fold increase in the synthesis of diacylglycerol, respectively. MGAT2 expressed in mammalian cells can catalyze the acylation of rac-1-, sn-2-, and sn-3 monoacylglycerols, and the enzyme prefers monoacylglycerols containing unsaturated fatty acyls as substrates. MGAT2 also demonstrates weak DGAT activity, which can be distinguished from its MGAT activity by detergent treatment that abolishes DGAT but not MGAT activity. We also analyzed the biochemical features of MGAT2 and demonstrated homogenate protein-, time-, and substrate concentration-dependent MGAT enzyme activity in transiently transfected COS-7 cells. Northern blot analysis indicates that the mouse MGAT2 is most abundantly expressed in the small intestine, suggesting that MGAT2 may play an important role in dietary fat absorption. PMID- 12576480 TI - Epigenomic stress response. Knockdown of DNA methyltransferase 1 triggers an intra-S-phase arrest of DNA replication and induction of stress response genes. AB - The DNA methylation pattern is an important component of the epigenome that regulates and maintains gene expression programs. In this paper, we test the hypothesis that vertebrate cells possess mechanisms protecting them from epigenomic stress similar to DNA damage checkpoints. We show that knockdown of DNMT1 (DNA methyltransferase 1) by an antisense oligonucleotide triggers an intra S-phase arrest of DNA replication that is not observed with control oligonucleotide. The cells are arrested at different positions throughout the S phase of the cell cycle, suggesting that this response is not specific to distinct classes of origins of replication. The intra-S-phase arrest of DNA replication is proposed to protect the genome from extensive DNA demethylation that could come about by replication in the absence of DNMT1. This protective mechanism is not induced by 5-aza-2'-deoxycytidine, a nucleoside analog that inhibits DNA methylation by trapping DNMT1 in the progressing replication fork, but does not reduce de novo synthesis of DNMT1. Our data therefore suggest that the intra-S-phase arrest is triggered by a reduction in DNMT1 and not by demethylation of DNA. DNMT1 knockdown also leads to an induction of a set of genes that are implicated in genotoxic stress response such as NF-kappaB, JunB, ATF-3, and GADD45beta (growth arrest DNA damage 45beta gene). Based on these data, we suggest that this stress response mechanism evolved to guard against buildup of DNA methylation errors and to coordinate inheritance of genomic and epigenomic information. PMID- 12576481 TI - Cross-talk between calpain and caspase proteolytic systems during neuronal apoptosis. AB - Cross-talk between calpain and caspase proteolytic systems has complicated efforts to determine their distinct roles in apoptotic cell death. This study examined the effect of overexpressing calpastatin, the specific endogenous calpain inhibitor, on the activity of the two proteolytic systems following an apoptotic stimulus. Human SH-SY5Y neuroblastoma cells were stably transfected with full-length human calpastatin cDNA resulting in 20-fold overexpression based on Western blot and 5-fold greater calpain inhibitory activity in cell extracts. Wild type and calpastatin overexpressing (CST1) cells were neuronally differentiated and apoptosis-induced with staurosporine (0.1-1.0 microm). Calpastatin overexpression decreased calpain activation, increased caspase-3-like activity, and accelerated the appearance of apoptotic nuclear morphology. Following 0.1-0.2 microm staurosporine, plasma membrane integrity based on calcein-acetoxymethyl fluorescence was significantly greater at 24 h in differentiated CST1 compared with differentiated wild type cells. However, this protective effect was lost at higher staurosporine doses (0.5-1.0 microm), which resulted in pronounced caspase-mediated degradation of the overexpressed calpastatin. These results suggest a dual role for calpains during neuronal apoptosis. In the early execution phase, calpain down-regulates caspase-3-like activity and slows progression of apoptotic nuclear morphology. Subsequent calpain activity, facilitated by caspase-mediated degradation of calpastatin, contributes to plasma membrane disruption and secondary necrosis. PMID- 12576482 TI - Expression of angiogenic factor Cyr61 during neuronal cell death via the activation of c-Jun N-terminal kinase and serum response factor. AB - The immediate early gene, cyr61, is transcriptionally activated within minutes by serum and serum growth factors. The encoded Cyr61 protein is secreted into the extracellular matrix and promotes cell adhesion and migration. In this study, we sought to examine the expression profile of cyr61 gene during neuronal cell death induced by various toxic stimuli and the mechanisms involved. Our data show that toxic stimuli, such as etoposide, significantly increased cyr61 mRNA levels in immortalized hippocampal progenitor (H19-7) cells. Cyr61 transcriptional activation was corroborated at the protein level as well. To identify the upstream signaling cascades involved in cyr61 gene induction, the blocking effect of either JNK or p38 kinase-signaling pathway on cyr61 induction in response to etoposide was tested. Transfection of the cells with a kinase-deficient mutant MEKK, an upstream activator of JNK, significantly decreased the cyr61 expression induced by etoposide. In contrast, cyr61 mRNA levels did not change after pretreatment with SB203580, the p38 kinase inhibitor. When the induction of cyr61 was tested by using several of its deleted promoters driving the expression of reporter gene, the promoter activation occurred primarily within the region containing an SRE-like CArG box. In addition, the SRF, which binds to the CArG site, was directly phosphorylated by active JNK. Furthermore, the blockade of cyr61 gene expression using an antisense encoding cyr61 sequence significantly inhibited the cell death induced by etoposide. Overall, these results suggest that the induction of the immediate early gene, cyr61, is important for neuronal cell death in the central nervous system hippocampal progenitor cells, and JNK activation, but not of p38, as well as the subsequent SRF phosphorylation are involved in cyr61 gene induction. PMID- 12576483 TI - Interaction of the tyrosine kinase Pyk2 with the N-methyl-D-aspartate receptor complex via the Src homology 3 domains of PSD-95 and SAP102. AB - The protein-tyrosine kinase Pyk2/CAKbeta/CADTK is a key activator of Src in many cells. At hippocampal synapses, induction of long term potentiation requires the Pyk2/Src signaling pathway, which up-regulates the activity of N-methyl-d aspartate-type glutamate receptors. Because localization of protein kinases close to their substrates is crucial for effective phosphorylation, we investigated how Pyk2 might be recruited to the N-methyl-d-aspartate receptor complex. This interaction is mediated by PSD-95 and its homolog SAP102. Both proteins colocalize with Pyk2 at postsynaptic dendritic spines in the cerebral cortex. The proline-rich regions in the C-terminal half of Pyk2 bind to the SH3 domain of PSD 95 and SAP102. The SH3 and guanylate kinase homology (GK) domain of PSD-95 and SAP102 interact intramolecularly, but the physiological significance of this interaction has been unclear. We show that Pyk2 effectively binds to the Src homology 3 (SH3) domain of SAP102 only when the GK domain is removed from the SH3 domain. Characterization of PSD-95 and SAP102 as adaptor proteins for Pyk2 fills a critical gap in the understanding of the spatial organization of the Pyk2-Src signaling pathway at the postsynaptic site and reveals a physiological function of the intramolecular SH3-GK domain interaction in SAP102. PMID- 12576484 TI - Endothelial cell overexpression of fas ligand attenuates ischemia-reperfusion injury in the heart. AB - Fas ligand (FasL) is a member of tumor necrosis factor family that induces apoptosis in target cells that express Fas. The function of FasL during inflammation remains controversial. In this study, we examined the role of vascular endothelial FasL during acute myocardial ischemia-reperfusion that is closely associated with inflammation. Transgenic mouse lines were established that overexpress human FasL on endothelium under the control of the vascular endothelial cadherin promoter. Expression of FasL transgene was detected at both mRNA and protein levels, and functional transgene-encoded FasL protein was specifically expressed on the surface of vascular endothelial cells. Transgenic mice developed normally and had normal hearts. When subjected to 30 min of myocardial ischemia and 72 h of reperfusion, myocardial infarct size was reduced by 42% in the transgenic mice compared with nontransgenic littermates (p < 0.05). Moreover, hemodynamic data demonstrated that transgenic hearts performed better following ischemia and reperfusion compared with nontransgenic hearts. Myocardial neutrophil infiltration was reduced by 54% after 6 h of reperfusion in transgenic hearts (p < 0.01). Neutrophil depletion prior to ischemia-reperfusion injury led to smaller infarcts that were not different between transgenic and nontransgenic mice, suggesting that endothelial FasL may attenuate ischemia-reperfusion injury by abating the inflammatory response. These results indicate that vascular endothelial FasL may exert potent anti-inflammatory actions in the setting of myocardial ischemia-reperfusion injury. PMID- 12576485 TI - Impairment of the DNA binding activity of the TATA-binding protein renders the transcriptional function of Rvb2p/Tih2p, the yeast RuvB-like protein, essential for cell growth. AB - In Saccharomyces cerevisiae, two highly conserved proteins, Rvb1p/Tih1p and Rvb2p/Tih2p, have been demonstrated to be major components of the chromatin remodeling INO80 complex. The mammalian orthologues of these two proteins have been shown to physically associate with the TATA-binding protein (TBP) in vitro but not clearly in vivo. Here we show that yeast proteins interact with TBP under both conditions. To assess the functional importance of these interactions, we examined the effect of mutating both TIH2/RVB2 and SPT15, which encodes TBP, on yeast cell growth. Intriguingly, only those spt15 mutations that affected the ability of TBP to bind to the TATA box caused synthetic growth defects in a tih2 ts160 background. This suggests that Tih2p might be important in recruiting TBP to the promoter. A DNA microarray technique was used to identify genes differentially expressed in the tih2-ts160 strain grown at the restrictive temperature. Only 34 genes were significantly and reproducibly affected; some up regulated and others down-regulated. We compared the transcription of several of these Tih2p target genes in both wild type and various mutant backgrounds. We found that the transcription of some genes depends on functions possessed by both Tih2p and TBP and that these functions are substantially impaired in the spt15/tih2-ts160 double mutants that confer synthetic growth defects. PMID- 12576486 TI - Thyroid hormone receptor-specific interactions with steroid receptor coactivator 1 in the pituitary. AB - Steroid receptor coactivator-1 (SRC-1) is a transcription cofactor that enhances the hormone-dependent action mediated by the thyroid hormone (TH) receptor (TR) as well as other nuclear receptors. However, it is not known whether the SRC-1 mediated activation of TH-regulated gene transcription is TR isoform specific in the pituitary. We generated mice that were deficient in TRalpha and SRC-1 (TRalpha(0/0)SRC-1(-/-)), as well in TRbeta and SRC-1 (TRbeta(-/-)SRC-1(-/-)), and thyroid function tests and effects of TH deprivation and TH treatment were compared with wild-type mice or mice with deletion of either TRs or SRC-1 alone. We have shown that 1) TRbeta(-/-)SRC-1(-/-) mice demonstrate more severe TH resistance than either the SRC-1(-/-) or TRbeta(-/-) mice; the additive effect indicates that SRC-1 has an independent role in TH action over that of TRbeta; 2) SRC-1 facilitates TRbeta and TRalpha-mediated down-regulation of TSH, as TRalpha(0/0)SRC-1(-/-) mice demonstrate TH resistance rather than hypersensitivity as seen in TRalpha(0/0)mice; and 3) a compensatory increase in SRC-1 expression is associated with the TH hypersensitivity seen in TRalpha deficient animals. We conclude that SRC-1 action in the pituitary mediates TH action via specific TR subtypes. PMID- 12576487 TI - Dimerization and signal transduction of the growth hormone receptor. AB - GH binding to cell surface-localized GH receptors (GHRs) induces a conformational change of the dimerized receptors, resulting in activation of Janus kinase 2 and downstream signaling pathways. Interactions between the extracellular subdomain 2 of adjacent GHR polypeptides result in a 500-A2 contact interface, which has previously been suggested to stabilize the GH-(GHR)2 complex. In this study, we investigated further the role of subdomain 2 in GHR function. Amino acids that participate in (e.g. aspartic acid 152, tyrosine 200, or serine 201) or lie close to (e.g. asparagine 143 or cysteine 241) the contact interface were mutated in rabbit GHR. Surprisingly, none of the mutations affected GHR dimerization, as demonstrated by coimmunoprecipitation of a truncated, epitope-tagged GHR. However, signal transduction of GHR(D152H), GHR(Y200D), and GHR(S201K) mutants was precluded. More insight into the molecular mechanism of the signaling defect was obtained when we examined the effect of the mutations on the integrity of the GH-(GHR)2 complex in a protease-protection assay. In contrast to wild-type GHR, GHR(N143K), and GHR(C241S), the GHR(D152H), GHR(Y200D), and GHR(S201K) mutants were not protected against protease digestion by GH, indicating that a structural change is prevented. Together, we provide new evidence for a critical role of aspartic acid 152, tyrosine 200, and serine 201 of the GHR contact interface in the GH-induced conformational change to a signaling-competent complex rather than in GHR dimerization. PMID- 12576488 TI - Marked potentiation of the dominant negative action of a mutant thyroid hormone receptor beta in mice by the ablation of one wild-type beta allele. AB - Mutations in the thyroid hormone receptor (TR) beta gene result in resistance to thyroid hormone (RTH), characterized by reduced sensitivity of tissues to thyroid hormone. To understand which physiological TR pathways are affected by mutant receptors, we crossed mice with a dominantly negative TRbeta mutation (TRbetaPV) with mice carrying a TRbeta null mutation (TRbeta(-/-)) to determine the consequences of the TRbetaPV mutation in the absence of wild-type TRbeta. TRbeta(PV/-) mice are distinct from TRbeta(+/-) mice that did not show abnormalities in thyroid function tests. TRbeta(PV/-) mice are also distinct from TRbeta(PV/+) and TRbeta(-/-) mice in that the latter shows mild dysfunction in the pituitary-thyroid axis, whereas the former exhibit very severe abnormalities, including extensive papillary hyperplasia of the thyroid epithelium, indistinguishable from that observed in TRbeta(PV/PV) mice. Similar to TRbeta(PV/PV) mice, TRbeta(PV/-) mice exhibited impairment in weight gain. Moreover, the abnormal regulation patterns of T3-target genes in the tissues of TRbeta(PV/-) and TRbeta(PV/PV) mice were strikingly similar. Using TR isoforms and PV-specific antibodies in gel shift assays, we found that in vivo, PV competed with TRalpha1 for binding to thyroid hormone response elements in TRbeta(PV/-) mice as effectively as in TRbeta(PV/PV) mice. Thus, the actions of mutant TRbeta are markedly potentiated by the ablation of the second TRbeta allele, suggesting that interference with wild-type TRalpha1-mediated gene regulation by mutant TRbeta leads to severe RTH. PMID- 12576489 TI - Pituitary corticotroph ontogeny and regulation in transgenic zebrafish. AB - We characterized zebrafish proopiomelanocortin (POMC) gene promoter, and sequence analysis revealed that the promoter contains regulatory elements conserved among vertebrate species. To monitor the ontogeny of the pituitary POMC lineage in living vertebrates, we generated transgenic zebrafish expressing green fluorescent protein (GFP) driven by the POMC promoter. Zebrafish POMC-GFP is first expressed asymmetrically as two bilateral groups of cells most anterior to the neural ridge midline at 18-20 h post fertilization (hpf). POMC-GFP-positive cells then fuse into a single-cell mass within the pituitary anlage after 24 hpf and subsequently organize as distinct anterior and posterior domains between 48 and 64 hpf. Immunohistochemical studies with ACTH and alphaMSH antisera showed that POMC-GFP was mainly targeted to both anterior and posterior pituitary corticotrophs, whereas posterior pituitary region melanotrophs did not express GFP. To determine in vivo zebrafish corticotroph responses, dexamethasone (10(-5) m) was added to live embryos, which selectively suppressed POMC-GFP expression in the anterior group of corticotrophs, suggesting a distinct domain that is responsive to glucocorticoid feedback. Transgenic zebrafish with specific POMC GFP expression in pituitary corticotrophs offers a powerful genetic system for in vivo study of vertebrate corticotroph lineage development. PMID- 12576490 TI - Domains of estrogen receptor alpha (ERalpha) required for ERalpha/Sp1-mediated activation of GC-rich promoters by estrogens and antiestrogens in breast cancer cells. AB - Estrogen receptor alpha (ERalpha)/Sp1 activation of GC-rich gene promoters in breast cancer cells is dependent, in part, on activation function 1 (AF1) of ERalpha, and this study investigates contributions of the DNA binding domain (C) and AF2 (DEF) regions of ERalpha on activation of ERalpha/Sp1. 17Beta-estradiol (E2) and the antiestrogens 4-hydroxytamoxifen and ICI 182,780 induced reporter gene activity in MCF-7 and MDA-MB-231 cells cotransfected with human or mouse ERalpha (hERalpha or MOR), but not ERbeta and GC-rich constructs containing three tandem Sp1 binding sites (pSp13) or other E2-responsive GC-rich promoters. Estrogen and antiestrogen activation of hERalpha/Sp1 was dependent on overlapping and different regions of the C, D, E, and F domains of ERalpha. Antiestrogen induced activation of hERalpha/Sp1 was lost using hERalpha mutants deleted in zinc finger 1 [amino acids (aa) 185-205], zinc finger 2 (aa 218-245), and the hinge/helix 1 (aa 265-330) domains. In contrast with antiestrogens, E2-dependent activation of hERalpha/Sp1 required the C-terminal F domain (aa 579-595), which contains a beta-strand structural motif. Moreover, in peptide competition experiments overexpression of a C-terminal (aa 575-595) F domain peptide specifically blocked E2-dependent activation of hERalpha/Sp1, suggesting that F domain interactions with nuclear cofactors are required for ERalpha/Sp1 action. PMID- 12576491 TI - Effect of 1-chloro-2,4-dinitrobenzene on K+ transport in normal and sickle human red blood cells. AB - 1-Chloro-2,4-dinitrobenzene (CDNB), which causes oxidative stress through depletion of reduced glutathione (GSH), increases the passive K+ permeability of red cells. In this paper, we investigated the effects of CDNB (1 mM) on the activities of the K+-Cl- cotransporter (KCC; measured as Cl--dependent K+ influx) and the Gardos channel (taken as clotrimazole-sensitive K+ influx, 5 microM) in human red cells, using 86Rb+ as a K+ congener. 45Ca2+ was used to study passive Ca2+ entry and active Ca2+ efflux via the plasma membrane Ca2+ pump. Both the Gardos channel and KCC were stimulated in both normal and sickle red cells. In sickle cells, stimulation of KCC was similar in oxygenated and deoxygenated cells; that of the Gardos channel was greater in deoxygenated cells. In normal red cells, stimulation of both pathways was greater in oxygenated cells (by 4 +/- 1-fold; all means +/- S.E.M., n = 3). The effects on the Gardos channel were dependent on extracellular Ca2+ and were associated with inhibition of the plasma membrane Ca2+ pump (by 29 +/- 3 %, P < 0.01) and increased Ca2+ sensitivity of the channel (EC50 for [Ca2+]i reduced from 260 +/- 26 to 175 +/- 15 nM; P < 0.05). Cell volume, pHi, ATP levels and passive Ca2+ entry were not affected by CDNB. The effects on KCC were inhibited (93 +/- 6 %) by prior treatment with the protein phosphatase inhibitor calyculin A (100 nM) and were not additive with stimulation by N-ethylmaleimide (1 mM), regardless of the order of addition. These findings are therefore consistent with inhibition of a regulatory protein kinase, although stimulation of the conjugate protein phosphatase(s) may also occur. KCC stimulation was also Ca2+ dependent. These findings are important for understanding how GSH depletion alters membrane permeability and how to protect against red cell dehydration. PMID- 12576492 TI - Developmental changes in membrane excitability and morphology of neurons in the nucleus angularis of the chicken. AB - In order to understand how sound intensity information is extracted and processed in the auditory nuclei, we investigated the neuronal excitability in the nucleus angularis (NA) of the chicken (P0-5) and the chicken embryo (E16-21). In embryos, neurons fired basically in three patterns in response to current injections: the onset pattern (19 %), the tonic pattern (52 %) and the pause pattern (29 %). After hatching, neurons fired either in the tonic pattern (83 %) or in the onset pattern (17 %). In both pre- and post-hatch periods, multiple firing neurons (tonic and pause) increased the maximum rate of rise of the action potential 2.6 fold, the fall 3.9-fold, and the maximum firing frequency 4-fold, and shifted the threshold potential to be more negative. After hatching, the firing frequency of tonic neurons reached a maximum at about 650 Hz. Application of TEA (1 mM) reduced the firing frequency, broadened action potentials and reduced the maximum rate of fall, but the threshold current was not changed. Dendrotoxin-I (DTX, 100 nM) reduced the threshold current. Application of DTX induced the onset neuron to fire repetitively. Branching patterns of auditory nerve fibres (ANFs) in NA were visualized by labelling with 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (Di-I) placed within the cochlea. Di-I placed near the apex of the cochlea labelled the ventral part of the NA, and Di-I placed in the base labelled the dorso-lateral part. Tonic neurons labelled with biocytin extended dendrites in parallel with the projection of ANFs in the nucleus after hatching. ANF activity of a limited range of characteristic sound frequencies is thought to be extracted by tonic neurons and encoded into firing frequencies proportional to the strength of the input. PMID- 12576493 TI - Peripheral nerve injury alters excitatory synaptic transmission in lamina II of the rat dorsal horn. AB - Using the blind whole cell patch-clamp recording technique, we investigated peripheral nerve injury-induced changes in excitatory synaptic transmission to neurones in lamina II of the dorsal horn. Partial (i.e. chronic constriction injury (CCI) and spared nerve injury (SNI)) and complete (i.e. sciatic nerve transection (SNT)) peripheral nerve injury altered the mean threshold intensity for eliciting A fibre-mediated EPSCs in lamina II neurones. Following SNT and CCI, EPSC threshold was significantly decreased, but following SNI, EPSC threshold was increased (naive: 32 +/- 2 mu A, SNT: 22 +/- 2 mu A, CCI: 23 +/- 2 mu A, SNI: 49 +/- 4 mu A; P < 0.01, Student's unpaired t test). Despite this disparity between models, dorsal root compound action potential recordings revealed no significant difference in the conduction velocity or activation threshold of A beta and A delta fibres in naive, SNT, CCI and SNI rats. In addition to the changes in EPSC threshold, we also observed a shift in the distribution of EPSCs. In spinal cord slices from naive rats, polysynaptic A beta fibre-evoked EPSCs were observed in 24 % of lamina II neurones, monosynaptic A delta fibre EPSCs were observed in 34 % and polysynaptic A delta fibre EPSCs were observed in 7 %. Following SNT and CCI, the percentage of neurones with polysynaptic A beta fibre EPSCs increased to > or = 65 % of the sampled population, while the percentage of neurones with monosynaptic A delta fibre EPSCs decreased to < 10 %. The percentage of neurones with polysynaptic A delta fibre EPSCs was unchanged. In contrast, following SNI, A beta fibre EPSCs decreased in incidence while the percentage of neurones with polysynaptic A delta fibre EPSCs increased to 44 %. Similar to the other injury models, however, monosynaptic A delta fibre EPSCs decreased in frequency following SNI. Thus, excitatory synaptic transmission is subject to divergent plasticity in different peripheral nerve injury models, reflecting the complexity of responses to different forms of deafferentation. PMID- 12576494 TI - PSD-95 regulates NMDA receptors in developing cerebellar granule neurons of the rat. AB - We transfected a green fluorescent protein-tagged PSD-95 (PSD-95gfp) into cultured rat cerebellar granule cells (CGCs) to investigate the role of PSD-95 in excitatory synapse maturation. Cells were grown in low potassium to favour functional synapse formation in vitro. Transfected cells displayed clear clusters of PSD-95gfp, often at the extremities of the short dendritic trees. We recorded NMDA and AMPA miniature excitatory postsynaptic currents (NMDA- and AMPA-mESPCs) in the presence of TTX and bicuculline. At days in vitro (DIV) 7-8 PSD-95gfp transfected cells had NMDA-mEPSCs with faster decay and smaller amplitudes than matching controls. In contrast, AMPA-mEPSC frequencies and amplitudes were increased. Whole-cell current density and ifenprodil sensitivity were reduced in PSD-95gfp cells, indicating a reduction of NR2B subunits containing NMDA receptors. No changes were observed compared to control when cells were transfected with cDNA for PSD-95gfp with palmitoylation site mutations that prevent targeting to the synapse. Overexpression of the NMDA receptor NR2A subunit, but not the NR2B subunit, prevented NMDA-mEPSC amplitude reduction when cotransfected with PSD-95gfp. PSD-95gfp overexpression produced faster NMDA-mEPSC decay when transfected alone or with either NR2 subunit. Surface staining of the epitope-tagged NR2 subunits revealed that colocalization with PSD-95gfp was higher for flag-tagged NR2A subunit clusters than for flag-tagged NR2B subunit clusters. These data suggest that PSD-95 overexpression in CGCs favours synaptic maturation by allowing synaptic insertion of NR2A and depressing expression of NR2B subunits. PMID- 12576495 TI - Developmental maturation of passive electrical properties in retinal ganglion cells of rainbow trout. AB - We investigated the electrotonic and anatomical features of the dendritic arbor in developing retinal ganglion cells (RGCs). Cell anatomy was studied by filling individual cells with fluorescent, membrane-bound dyes and using computer assisted image reconstruction. Electrotonic properties were characterized through an analysis of charging membrane currents measured with tight-seal electrodes in the whole-cell mode. We studied developing RGCs in the peripheral growth zone (PGZ) of a fish retina. The PGZ presents a developmental time-line ranging from pluripotent, proliferating cells at the extreme edge, to mature, fully developed retina more centrally. In the PGZ, RGCs mature through three histologically distinct zones (in developmental sequence): bulge, transition and mature zones. In the most peripheral three-quarters of the bulge zone, cells have rounded somas, lack dendritic extensions and some are coupled so that membrane-bound dyes traverse from one cell to its immediate neighbours. In the more central quarter of the bulge, cells' dendrites are few, short and of limited branching. In the transition zone dendritic arbors becomes progressively more expansive and branched and we present a morphometric analysis of these changes. Regardless of the size and branching pattern of the developing RGC dendritic arbor, the ratio of the diameters of parent and progeny dendrites at any branching nodes is well described by Rall's 3/2 power law. Given this anatomical feature, the RGC passive electrical properties are well described by an equivalent electrical circuit consisting of an isopotential cell body in parallel with a single equivalent cylinder of finite length. We measured the values of the electrical parameters that define this equivalent circuit in bulge, transition and mature RGCs. As RGCs develop the electrical properties of their dendritic arbor change in an orderly and tightly regulated manner, not randomly. Electrically, dendritic arbors develop along either of two distinct modes, but only these modes: isoelectrotonic and isometric. In isoelectrotonic growth, electrotonic properties are constant regardless of the absolute dimensions of the dendritic arbor or its branching geometry. These cells maintain unvarying relative synaptic efficacy independently of the size or pattern of their dendritic arbor. In isometric growth, in contrast, electronic properties change, but the ratio of the changing electrotonic length to electrotonic diameter is constant. In these cells relative synaptic efficacy decreases linearly as dendrites extend. PMID- 12576496 TI - Na+ channel inactivation: a comparative study between pancreatic islet beta-cells and adrenal chromaffin cells in rat. AB - A comparative study was carried out on the inactivation of Na+ channels in two types of endocrine cells in rats, beta-cells and adrenal chromaffin cells (ACCs), using patch-clamp techniques. The beta-cells were very sensitive to hyperpolarization; the Na+ currents increased ninefold when the holding potential was shifted from -70 mV to -120 mV. ACCs were not sensitive to hyperpolarization. The half-inactivation voltages were -90 mV (rat beta-cells) and -62 mV (ACCs). The time constant for recovery from inactivation at -70 mV was 10.5 times slower in beta-cells (60 ms) than in ACCs (5.7 ms). The rate of Na+-channel inactivation at physiological resting potential was more than three times slower in beta-cells than in ACCs. Na+ influx through Na+ channels had no effect on the secretory machinery in rat beta-cells. However, these 'silent Na+ channels' could contribute to the generation of action potentials in some conditions, such as when the cell is hyperpolarized. It is concluded that the fractional availability of Na+ channels in beta-cells at a holding potential of -70 mV is about 15 % of that in ACCs. This value in rat beta-cells is larger than that observed in mouse (0 %), but is smaller than those observed in human or dog (90 %). PMID- 12576497 TI - Store-operated Ca2+ entry: dynamic interplay between endoplasmic reticulum, mitochondria and plasma membrane. AB - In eukaryotic cells, hormones and neurotransmitters that engage the phosphoinositide pathway evoke a biphasic increase in intracellular free Ca2+ concentration: an initial transient release of Ca2+ from intracellular stores is followed by a sustained phase of Ca2+ influx. This influx is generally store dependent and is required for controlling a host of Ca2+-dependent processes ranging from exocytosis to cell growth and proliferation. In many cell types, store-operated Ca2+ entry is manifest as a non-voltage-gated Ca2+ current called ICRAC (Ca2+ release-activated Ca2+ current). Just how store emptying activates CRAC channels remains unclear, and some of our recent experiments that address this issue will be described. No less important from a physiological perspective is the weak Ca2+ buffer paradox: whereas macroscopic (whole cell) ICRAC can be measured routinely in the presence of strong intracellular Ca2+ buffer, the current is generally not detectable under physiological conditions of weak buffering following store emptying with the second messenger InsP3. In this review, I describe some of our experiments aimed at understanding just why InsP3 is ineffective under these conditions and which lead us to conclude that respiring mitochondria are essential for the activation of ICRAC in weak intracellular Ca2+ buffer. Mitochondrial Ca2+ uptake also increases the dynamic range over which InsP3 functions as the second messenger that controls Ca2+ influx. Finally, we find that Ca2+-dependent slow inactivation of Ca2+ influx, a widespread but poorly understood phenomenon that helps shape the profile of an intracellular Ca2+ signal, is regulated by mitochondrial Ca2+ buffering. Thus, by enabling macroscopic store-operated Ca2+ current to activate and then by controlling its extent and duration, mitochondria play a crucial role in all stages of store-operated Ca2+ influx. Store-operated Ca2+ entry reflects therefore a dynamic interplay between endoplasmic reticulum, mitochondria and plasma membrane. PMID- 12576498 TI - Ca2+ phase waves: a basis for cellular pacemaking and long-range synchronicity in the guinea-pig gastric pylorus. AB - Ca2+ imaging and multiple microelectrode recording procedures were used to investigate a slow wave-like electrical rhythmicity in single bundle strips from the circular muscle layer of the guinea-pig gastric pylorus. The 'slow waves' (SWs) consisted of a pacemaker and regenerative component, with both potentials composed of more elementary events variously termed spontaneous transient depolarizations (STDs) or unitary potentials. STDs and SW pacemaker and regenerative potentials exhibited associated local and distributed Ca2+ transients, respectively. Ca2+ transients were often larger in cellular regions that exhibited higher basal Ca2+ indicator-associated fluorescence, typical of regions likely to contain intramuscular interstitial cells of Cajal (ICCIM). The emergence of rhythmicity arose through entrainment of STDs resulting in pacemaker Ca2+ transients and potentials, events that exhibited considerable spatial synchronicity. Application of ACh to strips exhibiting weak rhythmicity caused marked enhancement of SW synchronicity. SWs and underlying Ca2+ increases exhibited very high 'apparent conduction velocities' ('CVs') orders of magnitude greater than for sequentially conducting Ca2+ waves. Central interruption of either intercellular connectivity or inositol 1,4,5-trisphosphate receptor (IP3R) mediated store Ca2+ release in strips caused SWs at the two ends to run independently of each other, consistent with a coupled oscillator-based mechanism. Central inhibition of stores required much wider regions of blockade than inhibition of connectivity indicating that stores were voltage-coupled. Simulations, made using a conventional store array model but now including depolarization coupled to IP3R-mediated Ca2+ release, predicted the experimental findings. The linkage between membrane voltage and Ca2+ release provides a means for stores to interact as strongly coupled oscillators, resulting in the emergence of Ca2+ phase waves and associated pacemaker potentials. This distributed pacemaker triggers regenerative Ca2+ release and resultant SWs. PMID- 12576499 TI - State-dependent regulation of cortical blood flow and respiration in hamsters: response to hypercapnia during arousal from hibernation. AB - Hibernation is characterised by a global reduction of metabolism, body temperature and blood flow, while arousal from hibernation is achieved by the reversal of these processes. Our experiments were performed on Syrian hamsters that had been chronically implanted with a cortical thermocouple and an optical fibre over the contralateral cortex, and acutely implanted with thermocouples in the rectal, cheek pouch and interscapular brown adipose tissue (BAT). Measurements revealed large thermal gradients in the body of the arousing animals. Maximum whole-body metabolic rate, which was 2.4 times normal cenothermic resting metabolic rate, coincided not with rectal temperature but more closely with respiratory rate (RR) or BAT temperature. Regional cortical blood flow (rCBF), as measured by laser-Doppler flowmetry, changed in parallel with whole-body metabolic rate, peaking at 3.8 times the normal cenothermic resting levels, when rectal temperature was 15 degrees C. When BAT temperature was less than 25 degrees C, RR, rCBF and heart rate (HR) were decreased by breathing hypercapnic gas, but these parameters were unresponsive to hyperoxic gases. At cenothermia the RR and rCBF of anaesthetised hamsters was increased by exposure to hypercapnic gases. Exposure to hyperoxic gas decreased RR but had no effect on rCBF. The mechanisms regulating rCBF, HR and RR exhibit state-dependent sensitivities to hypercapnic and hyperoxic stimuli. The large increase in rCBF observed during arousal implies that cerebral autoregulation is temporarily suspended and suggests that hamsters effectively use endogenous mechanisms to minimise the pathology normally associated with dramatic increases in rCBF. PMID- 12576501 TI - Heterologous expression of wild-type and mutant beta-cardiac myosin changes the contractile kinetics of cultured mouse myotubes. AB - The properties of myosin expressed in muscle are a major determinant of muscle performance. In this study we used a novel approach to examine the functional impact of changes in myosin heavy chain (MHC) isoform expression, as well as the consequences of expressing the mutant MHC implicated in familial hypertrophic cardiomyopathy (FHC). Cultured mouse myoblasts that normally express fast embryonic myosin were untransfected, or stably transfected with a plasmid expressing either wild-type (cWT) or mutant (D778G or G741R) beta-cardiac myosin. After differentiation for 5-7 days, cWT or mutant beta-cardiac myosin was expressed at 25 % of total myosin in the myotube. We measured time-to-peak shortening (ttp), time for half-relaxation (t0.5), the maximum velocity of shortening (Vmax) at 1 Hz stimulation, and the tetanic fusion frequency. Expression of cWT beta-cardiac myosin significantly increased ttp and t0.5 and decreased the fusion frequency compared with untransfected myotubes. However, when we compared myotubes expressing mutant beta-cardiac myosin with those expressing cWT beta-cardiac myosin, we found that ttp and t0.5 were significantly decreased, and Vmax was increased for the D778G mutant, whereas ttp, t0.5 and Vmax were unchanged for the G741R mutant. The fusion frequency was increased for both mutant myosins. Our data support the conclusion that the impact of the slower myosin isoform dominates when both slow and fast isoforms are present. This work suggests that FHC associated with either D778G or G741R mutation in MHC is an 'energy cost' disease, but that the phenotype of D778G is more severe than that of G741R. PMID- 12576500 TI - The actin cytoskeleton and cytotoxic T lymphocytes: evidence for multiple roles that could affect granule exocytosis-dependent target cell killing. AB - One important mechanism cytotoxic T lymphocytes (CTLs) use to kill virus infected, transplanted or tumour targets is exocytosis of granules that contain cytotoxic agents such as perforin and granzymes. Granule exocytosis-dependent target cell killing is a complex process, involving initial T-cell receptor (TCR) dependent signalling that includes Ca2+ influx and activation of protein kinase C, shape changes that serve to bind the CTL to the target and, finally, exocytosis of lytic granules at the site of contact with the target cell. Although there is reason to propose that multiple steps in the lytic process could involve the actin cytoskeleton of CTLs, few studies have examined this issue, and those that have do not allow the specific step(s) involved to be determined. We have used the potent membrane-permeant actin cytoskeleton modifying drugs jasplakinolide and latrunculin A to investigate the actin dependence of defined processes that are expected to be important for granule exocytosis-dependent killing. Our results, obtained using TALL-104 human leukaemic CTLs as a model system, are consistent with the idea that a functional actin cytoskeleton is required for TCR/CD3-dependent signalling, for activation of store-dependent Ca2+ influx and for CTL shape changes. When cells were stimulated with solid-phase anti-CD3 antibodies, treatment with either jasplakinolide or latrunculin A abolished granule exocytosis. However, when cells were stimulated in a manner that bypasses TCR/CD3-dependent signalling, granule exocytosis was not significantly altered, suggesting that the actin cytoskeleton does not function as a barrier to exocytosis. PMID- 12576502 TI - Action potential conduction in the terminal arborisation of nociceptive C-fibre afferents. AB - Recordings of single human peroneal C-fibres and rat saphenous C-fibres confirm two different patterns of conduction at branching points. In general, an action potential (AP) arising from one terminal branch may be propagated not only centrally, but also antidromically into the other branches of the terminal arborisation. If a stimulus activates several converging branches of one unit, at each branching point only the AP arriving first from the simultaneously activated daughter branches will be propagated centrally, resetting the slower branches. However, occasionally a single electrical stimulus may evoke a double response in the parent axon. In this case, these two responses apparently originate from different terminal branches and require unidirectional conduction block to prevent the faster AP from invading and resetting the slower-conducting terminal. This conclusion is supported by the notion that when such a double response occurs, both responses immediately show additional activity-dependent slowing of the conduction velocity due to frequency increase in the parent axon (two spikes per stimulus, one from each of the two excited branches). A comparable discharge pattern in the stem axon can be induced by repetitive paired stimulation of one terminal branch. Then the slowing is induced by the doubled frequency along the whole nerve fibre including the terminal branch. Since in this case not only the stem axon, but also the terminal branches carry two spikes per pulse, activity dependent slowing is predictably more pronounced. Unidirectional block thus provides insight into the differential amount of activity-dependent slowing (and hence postexcitatory hyperpolarisation) in the stem axon and terminal branches of cutaneous C-fibres. This comparison reveals that more than two-thirds of the slowing can be attributed to the terminal branches, since it is two- to fourfold that observed during double stimulation as compared with the unidirectional block condition. This indicates that the terminal branches are equipped with membrane proteins that are different from those of the parent axon. PMID- 12576505 TI - Regulation of transbilayer plasma membrane phospholipid asymmetry. AB - Lipids in biological membranes are asymmetrically distributed across the bilayer; the amine-containing phospholipids are enriched on the cytoplasmic surface of the plasma membrane, while the choline-containing and sphingolipids are enriched on the outer surface. The maintenance of transbilayer lipid asymmetry is essential for normal membrane function, and disruption of this asymmetry is associated with cell activation or pathologic conditions. Lipid asymmetry is generated primarily by selective synthesis of lipids on one side of the membrane. Because passive lipid transbilayer diffusion is slow, a number of proteins have evolved to either dissipate or maintain this lipid gradient. These proteins fall into three classes: 1) cytofacially-directed, ATP-dependent transporters ("flippases"); 2) exofacially-directed, ATP-dependent transporters ("floppases"); and 3) bidirectional, ATP-independent transporters ("scramblases"). The flippase is highly selective for phosphatidylserine and functions to keep this lipid sequestered from the cell surface. Floppase activity has been associated with the ABC class of transmembrane transporters. Although they are primarily nonspecific, at least two members of this class display selectivity for their substrate lipid. Scramblases are inherently nonspecific and function to randomize the distribution of newly synthesized lipids in the endoplasmic reticulum or plasma membrane lipids in activated cells. It is the combined action of these proteins and the physical properties of the membrane bilayer that generate and maintain transbilayer lipid asymmetry. PMID- 12576503 TI - Distinct contributions of small and large conductance Ca2+-activated K+ channels to rat Purkinje neuron function. AB - The cerebellum is important for many aspects of behaviour, from posture maintenance and goal-oriented reaching movements to timing tasks and certain forms of learning. In every case, information flowing through the cerebellum passes through Purkinje neurons, which receive input from the two primary cerebellar afferents and generate continuous streams of action potentials that constitute the sole output from the cerebellar cortex to the deep nuclei. The tonic firing behaviour observed in Purkinje neurons in vivo is maintained in brain slices even when synaptic inputs are blocked, suggesting that Purkinje neuron activity relies to a significant extent on intrinsic conductances. Previous research has suggested that the interplay between Ca2+ currents and Ca2+ activated K+ channels (KCa channels) is important for Purkinje cell activity, but how many different KCa channel types are present and what each channel type contributes to cell behaviour remains unclear. In order to better understand the ionic mechanisms that control the behaviour of these neurons, we investigated the effects of different Ca2+ channel and KCa channel antagonists on Purkinje neurons in acute slices of rat cerebellum. Our data show that Ca2+ entering through P type voltage-gated Ca2+ channels activates both small-conductance (SK) and large conductance (BK) KCa channels. SK channels play a role in setting the intrinsic firing frequency, while BK channels regulate action potential shape and may contribute to the unique climbing fibre response. PMID- 12576506 TI - Telomerase immortalization upregulates Rab9 expression and restores LDL cholesterol egress from Niemann-Pick C1 late endosomes. AB - Niemann-Pick C (NPC) disease is a rare recessive lipidosis marked by excessive accumulation of LDL-derived free cholesterol and glycosphingolipids in the late endosomal-lysosomal (E-L) system. Here we report that ectopic expression of human telomerase reverse transcriptase (hTeRT) in human cells leads to an upregulation of the small GTPase Rab9 and its effector p40. Expression of hTeRT in NPC1 cells results in a correction of their cellular phenotype, including clearance of accumulated cholesterol from their E-L system. Specifically, in NPC1-TeRT cells, the transport of cholesterol from the E-L system to the plasma membrane is restored with a concomitant increase in cholesterol esterification. This effect is Rab9-specific since expression of Rab9 in untransformed NPC1 cells also leads to a reversal of their disease phenotype. These effects are also seen in normal TeRT-immortalized cells and it appears that TeRT expression leads to an increase in the transport of molecules, including cholesterol, from the E-L system, and may play a role in increasing cellular proliferation. These results suggest the existence of alternative endogenous therapeutic targets that can be modulated to reverse the NPC1 disease phenotype. PMID- 12576507 TI - Abnormal splicing of ABCA1 pre-mRNA in Tangier disease due to a IVS2 +5G>C mutation in ABCA1 gene. AB - Two point mutations of ABCA1 gene were found in a patient with Tangier disease (TD): i) G>C in intron 2 (IVS2 +5G>C) and ii) c.844 C>T in exon 9 (R282X). The IVS2 +5G>C mutation was also found in the brother of another deceased TD patient, but not in 78 controls and 33 subjects with low HDL. The IVS2 +5G>C mutation disrupts ABCA1 pre-mRNA splicing in fibroblasts, leading to three abnormal mRNAs: devoid of exon 2 (Ex2-/mRNA), exon 4 (Ex4-/mRNA), or both these exons (Ex2-/Ex4 /mRNA), each containing a translation initiation site. These mRNAs are expected either not to be translated or generate short peptides. To investigate the in vitro effect of IVS2 +5G>C mutation, we constructed two ABCA1 minigenes encompassing Ex1-Ex3 region, one with wild-type (WTgene) and the other with mutant (MTgene) intron 2. These minigenes were transfected into COS1 and NIH3T3, two cell lines with a different ABCA1 gene expression. In COS1 cells, WTgene pre mRNA was spliced correctly, while the splicing of MTgene pre-mRNA resulted in Ex2 /mRNA. In NIH3T3, no splicing of MTgene pre-mRNA was observed, whereas WTgene pre mRNA was spliced correctly. These results stress the complexity of ABCA1 pre-mRNA splicing in the presence of splice site mutations. PMID- 12576508 TI - Fenofibrate, a ligand for PPARalpha, inhibits aromatase cytochrome P450 expression in the ovary of mouse. AB - Peroxisome proliferator-activated receptors (PPARs) play important roles in the metabolic regulation of lipids including steroids. In this study, we investigated whether fenofibrate, a ligand for PPARalpha, could influence estrogen synthesis in vivo in the ovary of mice. As reported, chronic treatment of C57BL6/J female mice with various amounts of fenofibrate as a diet reduced the serum triglycerides level and induced hepatomegaly in a dose-dependent manner. Northern blot analyses using hepatic RNA confirmed the induction of classical PPARalpha target genes including acyl-CoA oxidase and lipoprotein lipase. The analyses using ovarian RNA revealed the suppression of gene expression for enzymes involved in steroidogenesis including CYP11A, CYP19, steroidogenic acute regulatory protein, and HDL receptor, but the CYP17 expression was evidently induced. Consistent with the suppression of CYP19 mRNA expression, the aromatase activity in the ovary was dose-dependently inhibited, resulting in significant decreases in the uterine size and bone mineral density. When PPARalpha null mice were treated with dietary fenofibrate, neither hepatomegaly nor inhibition of ovarian aromatase activity was observed, rather the activity was enhanced. These results demonstrate that fenofibrate inhibits ovarian estrogen synthesis by suppressing the mRNA expressions and that functional PPARalpha is indispensable for the inhibitory action of the agent in vivo. PMID- 12576509 TI - Arachidonic acid and prostacyclin signaling promote adipose tissue development: a human health concern? AB - High fat intake is associated with fat mass gain through fatty acid activation of peroxisome proliferator-activated receptors delta and gamma, which promote adipogenesis. We show herein that, compared to a combination of specific agonists to both receptors or to saturated, monounsaturated, and omega-3 polyunsaturated fatty acids, arachidonic acid (C20:4, omega-6) promoted substantially the differentiation of clonal preadipocytes. This effect was blocked by cyclooxygenase inhibitors and mimicked by carbacyclin, suggesting a role for the prostacyclin receptor and activation of the cyclic AMP-dependent pathways that regulate the expression of the CCAAT enhancer binding proteins beta and delta implicated in adipogenesis. During the pregnancy-lactation period, mother mice were fed either a high-fat diet rich in linoleic acid, a precursor of arachidonic acid (LO diet), or the same isocaloric diet enriched in linoleic acid and alpha linolenic acid (LO/LL diet). Body weight from weaning onwards, fat mass, epididymal fat pad weight, and adipocyte size at 8 weeks of age were higher with LO diet than with LO/LL diet. In contrast, prostacyclin receptor-deficient mice fed either diet were similar in this respect, indicating that the prostacyclin signaling contributes to adipose tissue development. These results raise the issue of the high content of linoleic acid of i) ingested lipids during pregnancy and lactation, and ii) formula milk and infant foods in relation to the epidemic of childhood obesity. PMID- 12576510 TI - PPARbeta regulates vitamin A metabolism-related gene expression in hepatic stellate cells undergoing activation. AB - Activation of cultured hepatic stellate cells correlated with an enhanced expression of proteins involved in uptake and storage of fatty acids (FA translocase CD36, Acyl-CoA synthetase 2) and retinol (cellular retinol binding protein type I, CRBP-I; lecithin:retinol acyltransferases, LRAT). The increased expression of CRBP-I and LRAT during hepatic stellate cells activation, both involved in retinol esterification, was in contrast with the simultaneous depletion of their typical lipid-vitamin A (vitA) reserves. Since hepatic stellate cells express high levels of peroxisome proliferator activated receptor beta (PPARbeta), which become further induced during transition into the activated phenotype, we investigated the potential role of PPARbeta in the regulation of these changes. Administration of L165041, a PPARbeta-specific agonist, further induced the expression of CD36, B-FABP, CRBP-I, and LRAT, whereas their expression was inhibited by antisense PPARbeta mRNA. PPARbeta-RXR dimers bound to CRBP-I promoter sequences. Our observations suggest that PPARbeta regulates the expression of these genes, and thus could play an important role in vitA storage. In vivo, we observed a striking association between the enhanced expression of PPARbeta and CRBP-I in activated myofibroblast-like hepatic stellate cells and the manifestation of vitA autofluorescent droplets in the fibrotic septa after injury with CCl4 or CCl4 in combination with retinol. PMID- 12576511 TI - Role of the hepatic ABCA1 transporter in modulating intrahepatic cholesterol and plasma HDL cholesterol concentrations. AB - The current model for reverse cholesterol transport proposes that HDL transports excess cholesterol derived primarily from peripheral cells to the liver for removal. However, recent studies in ABCA1 transgenic mice suggest that the liver itself may be a major source of HDL cholesterol (HDL-C). To directly investigate the hepatic contribution to plasma HDL-C levels, we generated an adenovirus (rABCA1-GFP-AdV) that targets expression of mouse ABCA1-GFP in vivo to the liver. Compared with mice injected with control AdV, infusion of rABCA1-GFP-AdV into C57Bl/6 mice resulted in increased expression of mouse ABCA1 mRNA and protein in the liver. ApoA-I-dependent cholesterol efflux was increased 2.6-fold in primary hepatocytes isolated 1 day after rABCA1-GFP-AdV infusion. Hepatic ABCA1 expression in C57Bl/6 mice (n = 15) raised baseline levels of TC, PL, FC, HDL-C, apoE, and apoA-I by 150-300% (P < 0.05 all). ABCA1 expression led to significant compensatory changes in expression of genes that increase hepatic cholesterol, including HMG-CoA reductase (3.5-fold), LDLr (2.1-fold), and LRP (5-fold) in the liver. These combined results demonstrate that ABCA1 plays a key role in hepatic cholesterol efflux, inducing pathways that modulate cholesterol homeostasis in the liver, and establish the liver as a major source of plasma HDL-C. PMID- 12576512 TI - Cloning of monkey RALDH1 and characterization of retinoid metabolism in monkey kidney proximal tubule cells. AB - All-trans and 9-cis retinoic acids function as ligands for retinoic acid receptors (RARs and RXRs), which are ligand-dependent transcription factors and play important roles in development and cellular differentiation. Several retinal dehydrogenases are likely to contribute to the production of all-trans and 9-cis RAs in vivo, but their respective roles in different tissues are still poorly characterized. We have previously characterized and cloned from kidney tissues the rat retinal dehydrogenase type 1 (RALDH1), which oxidizes all-trans and 9-cis retinal with high efficiency but is inactive with 13-cis retinal. Here we have characterized the retinal-oxidizing activity in monkey JTC12 cells, which are derived from kidney proximal tubules. In vitro assay of cell lysates revealed the presence of a NAD+-dependent dehydrogenase that catalyzed the oxidation of all trans, 9-cis, and 13-cis retinal. Northern blot analysis of JTC12 RNAs and cloning by reverse transcription-polymerase chain reaction demonstrated expression of a monkey homolog of RALDH1. Bacterially expressed JTC12 RALDH1 catalyzed conversion of all three retinal isomers, with a higher catalytic efficiency for 9-cis retinal than for all-trans and 13-cis retinal. Accordingly, live JTC12 produced 9-cis retinoic acid more efficiently than all-trans retinoic acid from their respective retinal precursors. Only metabolites corresponding to the same steric conformation were formed from 9-cis or all-trans retinal, indicating a lack of detectable isomerizing activity in JTC12 cells. PMID- 12576513 TI - Whole-body beta-oxidation of 18:2omega6 and 18:3omega3 in the pig varies markedly with weaning strategy and dietary 18:3omega3. AB - Segregated early weaning (SEW) into a cleaner nursery increases food intake and growth in pigs, presumably because of reduced immune stimulation compared with conventionally reared, nonsegregated pigs (NSW). The aim of the present study was to evaluate the oxidation of linoleic acid (18:2omega6) and alpha-linolenic acid (18:3omega3) in SEW and NSW pigs. Pigs consumed a control or high 18:3omega3 diet (omega6 PUFA/omega3 PUFA; 21.3 vs. 2.5, respectively) and were weaned at either 14 days old into a SEW nursery or at 21 days old into a conventional NSW nursery. The major acute-phase protein of pigs but not haptoglobin increased in 35-day-old NSW pigs. NSW pigs had 15-25% lower carcass 18:2omega6 and 20-30% lower carcass 18:3omega3 (% composition) at 49 days old. Between 35- and 49-days-old, NSW pigs had a higher whole-body oxidation of 18:2omega6 (40-120%) and 18:3omega3 (30 80%). The high 18:3omega3 diet decreased the whole-body oxidation of 18:2omega6 by 73% and of 18:3omega3 by 63% in NSW pigs. We conclude that moderately cleaner housing SEW significantly decreases 18:2omega6 and 18:3omega3 oxidation in pigs. PMID- 12576514 TI - pH6 antigen of Yersinia pestis interacts with plasma lipoproteins and cell membranes. AB - The bacterial pathogen Yersinia pestis expresses a potential adhesin, the pH6 antigen (pH6-Ag), which appears as fimbria-like structures after exposure of the bacteria to low pH. pH6-Ag was previously shown to agglutinate erythrocytes and to bind to certain galactocerebrosides. We demonstrate that purified pH6-Ag selectively binds to apolipoprotein B (apoB)-containing lipoproteins in human plasma, mainly LDL. Binding was not prevented by antibodies to apoB. pH6-Ag interacted also with liposomes and with a lipid emulsion, indicating that the lipid moiety of the lipoprotein was responsible for the interaction. Both apoB containing lipoproteins and liposomes prevented binding of pH6-Ag to THP-I monocyte-derived macrophages as well as pH6-Ag-mediated agglutination of erythrocytes. Binding of pH6-Ag to macrophages was not dependent on the presence of LDL receptors. Treatment of the cells with Triton X-100 or with methyl-beta cyclodextrin indicated that the binding of pH6-Ag was partly dependent on lipid rafts. We suggest that interaction of pH6-Ag with apoB-containing lipoproteins could be of importance for the establishment of Y. pestis infections. Binding of lipoproteins to the bacterial surface could prevent recognition of the pathogen by the host defence systems. This might be important for the ability of the pathogen to replicate in the susceptible host. PMID- 12576515 TI - SR-BI-directed HDL-cholesteryl ester hydrolysis. AB - We have examined the metabolic fate of HDL cholesteryl ester (CE) delivered to cells expressing scavenger receptor class B type I (SR-BI). Comparison of SR-BI with a related class B scavenger receptor, CD36, showed a greater uptake and a more rapid and extensive hydrolysis of HDL-CE when delivered by SR-BI. In addition, hydrolysis of HDL-CE delivered by both receptors was via a neutral CE hydrolase. These data indicate that SR-BI, but not CD36, can efficiently direct HDL-CE to a neutral CE hydrolytic pathway. In contrast, LDL-CE was delivered and hydrolyzed equally well by SR-BI and CD36. Hydrolysis of LDL-CE delivered by SR BI was via a neutral CE hydrolase but that delivered by CD36 occurred via an acidic CE hydrolase, indicating that SR-BI and CD36 deliver LDL-CE to different metabolic pathways. Comparison of inhibitor sensitivities in Y1-BS1 adrenal, Fu5AH hepatoma, and transfected cells suggests that hydrolysis of HDL-CE delivered by SR-BI occurs via cell type-specific neutral CE hydrolases. Furthermore, HDL-CE hydrolytic activity was recovered in a membrane fraction of Y1-BS1 cells. These findings suggest that SR-BI efficiently delivers HDL-CE to a metabolically active membrane compartment where CE is hydrolyzed by a neutral CE hydrolase. PMID- 12576516 TI - Effect of GM2 activator protein on the enzymatic hydrolysis of phospholipids and sphingomyelin. AB - GM2 activator protein (GM2AP) is a specific protein cofactor that stimulates the enzymatic hydrolysis of the GalNAc from GM2, a sialic acid containing glycosphingolipid, both in vitro and in lysosomes. While phospholipids together with glycosphingolipids are important membrane constituents, little is known about the possible effect of GM2AP on the hydrolysis of phospholipids. Several recent reports suggest that GM2AP might have functions other than stimulating the conversion of GM2 into GM3 by beta-hexosaminidase A, such as inhibiting the activity of platelet activating factor and enhancing the degradation of phosphatidylcholine by phospholipase D (PLD). We therefore examined the effect of GM2AP on the in vitro hydrolyses of a number of phospholipids and sphingomyelin by microbial (Streptomyces chromofuscus) and plant (cabbage) PLD. GM2AP, at the concentration as low as 1.08 microM (1 microg/50 microl) was found to inhibit about 70% of the hydrolyses of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol by PLD, whereas the same concentration of GM2AP only inhibited about 20-25% of the hydrolysis of sphingomyelin by sphingomyelinase and had no effect on the hydrolysis of sphingosylphosphorylcholine by PLD. Thus, GM2AP exerts strong and broad inhibitory effects on the hydrolysis of phospholipids carried out by plant and microbial PLDs. High ammonium sulfate concentration (1.6 M or 21.1%) masks this inhibitory effect, possibly due to the alteration of the ionic property of GM2AP. PMID- 12576517 TI - Characterization of specifically oxidized apolipoproteins in mildly oxidized high density lipoprotein. AB - Atherosclerosis is a state of heightened oxidative stress. Oxidized LDL is present in atherosclerotic lesions and used as marker for coronary artery disease, although in human lesions lipids associated with HDL are as oxidized as those of LDL. Here we investigated specific changes occurring to apolipoprotein A I (apoA-I) and apoA-II, as isolated HDL and human plasma undergo mild, chemically induced oxidation, or autoxidation. During such oxidation, Met residues in apoA-I and apoA-II become selectively and consecutively oxidized to their respective Met sulfoxide (MetO) forms that can be separated by HPLC. Placing plasma at -20 degrees C prevents autoxidation, whereas metal chelators and butylated hydroxytoluene offer partial protection. Independent of the oxidation conditions, apoA-I and apoA-II (dimer) with two MetO residues accumulate as relatively stable oxidation products. Compared to controls, serum samples from subjects with the endothelial cell nitric oxide synthase a/b genotype that is associated with increased coronary artery disease contain increased concentrations of apoA-I with two MetO residues. Our results show that during the early stages, oxidation of HDL gives rise to specifically oxidized forms of apoA-I and apoA-II, some of which may be useful markers of in vivo HDL oxidation, and hence potentially atherosclerosis. PMID- 12576518 TI - SREBP-1 integrates the actions of thyroid hormone, insulin, cAMP, and medium chain fatty acids on ACCalpha transcription in hepatocytes. AB - In chick embryo hepatocytes, activation of acetyl-CoA carboxylase-alpha (ACCalpha) transcription by 3,5,3'-triiodothyronine (T3) is mediated by a cis acting regulatory unit (-101 to -71 bp) that binds the nuclear T3 receptor (TR) and sterol regulatory element-binding protein-1 (SREBP-1). SREBP-1 directly interacts with TR on the ACCalpha gene to enhance T3-induced transcription. Here, we show that treating hepatocytes with T3 or insulin stimulates a 4-fold increase in the concentration of the mature, active form of SREBP-1. When T3 and insulin are added together, a 7-fold increase in the mature SREBP-1 concentration is observed. Time course studies indicate that the T3-induced increase in mature SREBP-1 abundance is closely associated with changes in ACCalpha transcription and that the mechanism mediating the effect of T3 on mature SREBP-1 is distinct from that mediating the effect of insulin. Transfection analyses indicate that inhibition of ACCalpha transcription by cAMP or hexanoate is mediated by ACCalpha sequences between -101 and -71 bp. Treatment with cAMP or hexanoate suppresses the increase in mature SREBP-1 abundance caused by T3 and insulin. These results establish a new interaction between the SREBP-1 and TR signaling pathways and provide evidence that SREBP-1 plays an active role in mediating the effects of T3, insulin, cAMP, and hexanoate on ACCalpha transcription. PMID- 12576519 TI - A low fish oil inhibits SREBP-1 proteolytic cascade, while a high-fish-oil feeding decreases SREBP-1 mRNA in mice liver: relationship to anti-obesity. AB - Rodents fed fish oil showed less obesity with a reduction of triglyceride synthesis in liver, relative to other dietary oils, along with a decrease of mature form of sterol regulatory element binding protein-1 (SREBP-1) and activation of peroxisome proliferator-activated receptor alpha (PPARalpha). Decrease of mature SREBP-1 protein by fish oil feeding was due to either inhibition of SREBP-1 proteolytic cascade or to decrease of its mRNA. To clarify its mechanism and relation to antiobesity effect, mice were fed fish oil in a range from 10 to 60 energy percent (en%). Fish oil feeding decreased body weight and fat mass in a dose-dependent manner, in parallel with PPARalpha activation and a decrease of SREBP-1 mRNA. However, compared with 0 en% fish oil feeding, 10 en% fish oil feeding decreased mature SREBP-1 protein by 50% with concomitant decreases of lipogenic genes, while precursor SREBP-1 protein rather increased by 1.3-fold. These data suggest that physiological doses of fish oil feeding effectively decrease expression of liver lipogenic enzymes by inhibiting SREBP-1 proteolytic cascade, while substantial decrease of SREBP-1 expression is observed in its pharmacological doses, and that activation of PPARalpha rather than SREBP 1 decrease might be related to the antiobesity effect of fish oil feeding. PMID- 12576520 TI - Modulation by flavonoids of PAF and related phospholipids in endothelial cells during oxidative stress. AB - PAF-dependent transacetylase (TA) modifies the functions of platelet-activating factor (PAF), a potent inflammatory lipid, either by transferring the acetyl group from PAF to lysophospholipids (TAL activity), or to sphingosine (TAS activity) or by hydrolyzing PAF (acetylhydrolase activity). In stimulated endothelial cells (EC), TAL activity contributes to the synthesis of acyl-PAF, an acyl analog of PAF, that antagonizes PAF functions and is regulated by the cellular redox state. In this study, we investigated the possible involvement of TA in the flavonoid antioxidant mechanism(s) during oxidative stress in EC induced by hydrogen peroxide. The treatment of EC with H2O2 resulted in 4-fold increase of the acetyl-CoA acetyltransferase activity (AT), that is responsible for PAF biosynthesis, while the TAL activity increased only by 53%. However, the preincubation of H2O2-treated EC with the flavonoids hesperedin, naringin, and quercetin strongly inhibited AT activity and activated TAL by 290%, 340%, and 250%, respectively. The induction of TAL activity resulted in enhanced biosynthesis of 1-acyl-2-[3H]acetyl-PAF in intact EC and was related to the flavonoid structure. These findings suggest that TAL is involved in the flavonoid anti-inflammatory action by enhancing the production of acyl-PAF. PMID- 12576521 TI - Down-regulation of acyl-CoA oxidase gene expression and increased NF-kappaB activity in etomoxir-induced cardiac hypertrophy. AB - Activation of nuclear factor-kappaB (NF-kappaB) is required for hypertrophic growth of cardiomyocytes. Etomoxir is an irreversible inhibitor of carnitine palmitoyltransferase I (CPT-I) that activates peroxisome proliferator-activated receptor alpha (PPARalpha) and induces cardiac hypertrophy through an unknown mechanism. We studied the mRNA expression of genes involved in fatty acid oxidation in the heart of mice treated for 1 or 10 days with etomoxir (100 mg/kg/day). Etomoxir administration for 1 day significantly increased (4.4-fold induction) the mRNA expression of acyl-CoA oxidase (ACO), which catalyzes the rate-limiting step in peroxisomal beta-oxidation. In contrast, etomoxir treatment for 10 days dramatically decreased ACO mRNA levels by 96%. The reduction in ACO expression in the hearts of 10-day etomoxir-treated mice was accompanied by an increase in the mRNA expression of the antioxidant enzyme glutathione peroxidase and the cardiac marker of oxidative stress bax. Moreover, the activity of the redox-regulated transcription factor NF-kappaB was increased in heart after 10 days of etomoxir treatment. Overall, the findings here presented show that etomoxir treatment may induce cardiac hypertrophy via increased cellular oxidative stress and NF-kappaB activation. PMID- 12576522 TI - Hepatic overexpression of sterol carrier protein-2 inhibits VLDL production and reciprocally enhances biliary lipid secretion. AB - We examined in vivo a role for sterol carrier protein-2 (SCP-2) in the regulation of lipid secretion across the hepatic sinusoidal and canalicular membranes. Recombinant adenovirus Ad.rSCP2 was used to overexpress SCP-2 in livers of mice. We determined plasma, hepatic, and biliary lipid concentrations; hepatic fatty acid (FA) and cholesterol synthesis; hepatic and biliary phosphatidylcholine (PC) molecular species; and VLDL triglyceride production. In Ad.rSCP2 mice, there was marked inhibition of hepatic fatty acids and cholesterol synthesis to <62% of control mice. Hepatic triglyceride contents were decreased, while cholesterol and phospholipids concentrations were elevated in Ad.rSCP2 mice. Hepatic VLDL triglyceride production fell in Ad.rSCP2 mice to 39% of control values. As expected, biliary cholesterol, phospholipids, bile acids outputs, and biliary PC hydrophobic index were significantly increased in Ad.rSCP2 mice. These studies indicate that SCP-2 overexpression in the liver markedly inhibits lipid synthesis as well as VLDL production, and alters hepatic lipid contents. In contrast, SCP-2 increased biliary lipid secretion and the proportion of hydrophobic PC molecular species in bile. These effects suggest a key regulatory role for SCP-2 in hepatic lipid metabolism and the existence of a reciprocal relationship between the fluxes of lipids across the sinusoidal and canalicular membranes. PMID- 12576523 TI - Hyperlipidemia in APOE2 transgenic mice is ameliorated by a truncated apoE variant lacking the C-terminal domain. AB - Familial dysbetalipoproteinemia associated with the apolipoprotein E2 (APOE2) genotype is a recessive disorder with low penetrance. We have investigated whether additional expression of full-length APOE3, APOE4, or a truncated variant of APOE4 (APOE4-202) can reduce APOE2- associated hyperlipidemia. This was achieved using adenovirus-mediated gene transfer to mice transgenic for human APOE2 and deficient for endogenous Apoe (APOE2.Apoe-/- mice). The hyperlipidemia of APOE2.Apoe-/- mice was readily aggravated by APOE3 and APOE4 overexpression. Only a very low dose of APOE4 adenovirus was capable of reducing the serum cholesterol and triglyceride (TG) levels. Expression of higher doses of APOE4 was associated with an increased VLDL-TG production rate and the accumulation of TG rich VLDL in the circulation. In contrast, a high dose of adenovirus carrying APOE4-202 reduced both the cholesterol and TG levels in APOE2.Apoe-/- mice. Despite the absence of the C-terminal lipid-binding domain, APOE4-202 is apparently capable of binding to lipoproteins and mediating hepatic uptake. Moreover, overexpression of APOE4-202 in APOE2.Apoe-/- mice does not aggravate their hypertriglyceridemia. These results extend our previous analyses of APOE4 202 expression in Apoe-/- mice and demonstrate that apoE4-202 functions even in the presence of clearance-defective apoE2. Thus, apoE4-202 is a safe and efficient candidate for future therapeutic applications. PMID- 12576524 TI - Etomoxir mediates differential metabolic channeling of fatty acid and glycerol precursors into cardiolipin in H9c2 cells. AB - We examined the effect of etomoxir treatment on de novo cardiolipin (CL) biosynthesis in H9c2 cardiac myoblast cells. Etomoxir treatment did not affect the activities of the CL biosynthetic and remodeling enzymes but caused a reduction in [1-14C]palmitic acid or [1-14C]oleic acid incorporation into CL. The mechanism was a decrease in fatty acid flux through the de novo pathway of CL biosynthesis via a redirection of lipid synthesis toward 1,2-diacyl-sn-glycerol utilizing reactions mediated by a 35% increase (P < 0.05) in membrane phosphatidate phosphohydrolase activity. In contrast, etomoxir treatment increased [1,3-3H]glycerol incorporation into CL. The mechanism was a 33% increase (P < 0.05) in glycerol kinase activity, which produced an increased glycerol flux through the de novo pathway of CL biosynthesis. Etomoxir treatment inhibited 1,2-diacyl-sn-glycerol acyltransferase activity by 81% (P < 0.05), thereby channeling both glycerol and fatty acid away from 1,2,3-triacyl-sn glycerol utilization toward phosphatidylcholine and phosphatidylethanolamine biosynthesis. In contrast, etomoxir inhibited myo-[3H]inositol incorporation into phosphatidylinositol and the mechanism was an inhibition in inositol uptake. Etomoxir did not affect [3H]serine uptake but resulted in an increased formation of phosphatidylethanolamine derived from phosphatidylserine. The results indicate that etomoxir treatment has diverse effects on de novo glycerolipid biosynthesis from various metabolic precursors. In addition, etomoxir mediates a distinct and differential metabolic channeling of glycerol and fatty acid precursors into CL. PMID- 12576525 TI - Role of cyclooxygenases COX-1 and COX-2 in modulating adipogenesis in 3T3-L1 cells. AB - Cyclooxygenase (COX) catalyses the rate-limiting step of prostanoid biosynthesis. Two COX isoforms have been identified, COX-1, the constitutive form, and COX-2, the inducible form. While COX-2 has been implicated in body fat regulation, the underlying cellular mechanism remains to be elucidated. The present study was undertaken to examine the potential role of COX in modulating adipogenesis and to dissect the relative contribution of the two isoenzymes in this process. COX-2 was found to be expressed in undifferentiated 3T3-L1 cells and down-regulated during differentiation, whereas the cellular level of COX-1 remained relatively constant. Abrogating the activity of either of these two isoenzymes by selective COX inhibitors accelerated cellular differentiation, suggesting that both COX isoenzymes negatively influenced differentiation. Tumor necrosis factor-alpha (TNFalpha) significantly up-regulated COX-2 expression ( approximately 2-fold) in differentiating 3T3-L1 cells, whereas similar effect was not observed with COX-1 expression. Abrogating the induced COX-2 activity reversed the TNFalpha-induced inhibition of differentiation by approximately 70%, implying a role for COX-2 in mediating TNFalpha signaling. Hence, both COX isoforms were involved in the negative modulation of adipocyte differentiation. COX-2 appeared to be the main isoform mediating at least part of the negative effects of TNFalpha. PMID- 12576526 TI - Biological and genetic determinants of serum apoC-III concentration: reference limits from the Stanislas Cohort. AB - Apolipoprotein C-III (apoC-III) is involved in triglycerides metabolism, and is therefore important for the pathogenesis of coronary heart diseases. However, to our knowledge serum apoC-III variation factors and reference limits have never been determined, so the aim of this study was to establish them and facilitate clinical usefulness. We measured serum apoC-III concentration of apparently healthy subjects of the Stanislas Cohort by an immunoturbidimetric method. Genetic polymorphisms within the APOC3, APOE, APOAIV, and LPL genes were determined by a multiplex PCR. Serum apoC-III concentration varied from 28.2 mg/l to 225.8 mg/l in the overall sample and between subjects variability was about 30%. Factors influencing apoC-III concentration were age, BMI in adult men, alcohol consumption in adults, oral contraceptive intake in women, the post pubescent status in boys. The APOC3 1100T allele in adult men and the APOC3 -455C allele in boys were associated with increased apoC-III concentration. The APOA4 360His allele was associated with decreased apoC-III concentration in women. We also established reference limits of serum apoC-III concentration according to age and gender. PMID- 12576528 TI - Precautionary principle encourages policies that protect human health and the environment in the face of uncertain risks. PMID- 12576529 TI - The heart of the precautionary principle in democracy. PMID- 12576530 TI - Industry challenges to the principle of prevention in public health: the precautionary principle in historical perspective. PMID- 12576531 TI - Anthrax: the precautionary principle goes postal. PMID- 12576532 TI - Science, precaution, and practice. PMID- 12576533 TI - Developing scientific and policy methods that support precautionary action in the face of uncertainty--the Institute of Medicine Committee on Agent Orange. AB - To be precautionary, decisions must be made to prevent the impacts of potentially harmful activities even though the nature and magnitude of harm have not been proven scientifically. The Institute of Medicine's Committee on the Health Effects in Vietnam Veterans of Exposures to Herbicides provides a novel example of science and policy structures that support precautionary action in the face of uncertainty. What makes this example unique is the clear set of precautionary decision rules that lowered the standard for evidence, which formed the basis for policy. These rules, established by Congress, strongly influenced the way scientific information was weighed and the subsequent compensation decisions. They encouraged committee members to think outside the confines of their disciplines and develop new tools and methods to fit their unique mandate. The result was a methodology, supported by strong institutional structures, that allowed scientists to discuss the evidence as a whole, reach decisions as a group, and clarify uncertainties. PMID- 12576534 TI - The precautionary principle and emerging biological risks: lessons from swine flu and HIV in blood products. AB - Two examples-the "swine flu affair" in 1976 and the emergence of HIV in the blood supply in the early 1980s-illustrate the difficulties of decision-making in public health. Both cases illustrate trade-offs between product risks and public health benefits, especially with regard to uncertainty in estimates of product risks, public health risks, and the benefits of prevention. The cases also illustrate the tendency of public health policy makers to go all the way or do nothing at all, rather than consider intermediate options that can be adapted as new information emerges. This review suggests three lessons for public health policy makers: (1) be open and honest about scientific uncertainty; (2) communicate with the public, even when the facts are not clear; and (3) consider intermediate, adaptable policy options, such as obtaining more information, thus reducing uncertainty, and building in decision points to reconsider initial policies. Underlying all of these lessons is the need to commission studies to resolve important uncertainties and increase the information base for public communication, and to review regulations and other policy options in the light of the new data that emerge. PMID- 12576535 TI - Transparent Democratic Foresight Strategies in the California EMF Program. AB - A California Department of Health Services program dealt with possible health effects from Electric and Magnetic Fields (EMF) from power lines. With the advice of stakeholders, and well before any risk determinations were made, transparent policy analyses about the power grid and schools asked the question, "How confident must one be of how big an effect before one would adopt cheap or expensive EMF avoidance measures?" A risk evaluation was carried out with features that promoted transparency. It was formatted to provide a policy-neutral "degree of certainty of causality" to adherents of utilitarian, environmental justice, and libertarian policy frameworks. Though the program had many features advocated by adherents of the precautionary principle, it might be better characterized as following "Transparent Democratic Foresight Strategies," since no single principle justifies the strategies used in this participatory program, and it examined the pros and cons of options but made no recommendations, precautionary or otherwise. PMID- 12576536 TI - Primary prevention and precaution in hazard identification in the NIEHS/NTP: body in the morgue approach. AB - In the spirit of stimulating reevaluation of the methods of public health science, this article explores the methods of cancer hazard identification at the National Toxicology Program (NTP) from the perspective of primary prevention and precaution. The NTP is a cooperative effort of three federal agencies: The National Institute of Environmental Health Sciences (NIEHS, the lead government institute); the National Institute for Occupational Health (NIOSH), in the Centers for Disease Control and Prevention; and the National Center for Toxicology Research (NCTR), in the Food and Drug Administration. NTP coordinates toxicological research and testing programs within the Department of Health and Human Services (DHHS), and through its annual Report on Carcinogens (RoC), identifies and characterizes cancer hazards-the first step in quantitative risk assessment-for the federal government. The foundation of NIEHS policies, for environmental health research, is quantitative risk assessment (QRA). The author examines the opportunities for primary prevention and precaution, and the extent to which the policies of NIEHS in general, and the NTP in particular, do and do not realize that potential. Special attention is paid to the issue of cancer hazard identification. Critical comments on the process of classifying carcinogens in the Ninth and Tenth Reports on Carcinogens are presented, based on the minutes of the Board of Scientific Counselors Subcommittee meetings. PMID- 12576537 TI - Moving further upstream: from toxics reduction to the precautionary principle. AB - Early policies to reduce the amount of toxic waste in the environment focused on cleaning up downstream sources of pollution, such as toxic disposal sites. Public attention in the 1980s encouraged both industry and government to develop an alternative to this command-and-control approach. This article describes the emergence of that alternative-pollution prevention-and its application in Massachusetts through the 1989 Toxics Use Reduction Act. Pollution prevention focuses on the sources of pollution, both metaphorically and physically, more upstream than its predecessors. The success of the Toxics Use Reduction Act in Massachusetts helped create an opportunity where an alternative pollution prevention paradigm could develop. That paradigm, the precautionary principle, is popular among environment activists because it focuses further upstream than pollution prevention by calling attention to the role the social construction of risk plays in decisions regarding the use of hazardous substances. The authors examine the evolution of the precautionary principle through an investigation of three major pathways in its development and expansion. The article concludes with a discussion of the increased potential for protecting public health and the environment afforded by this new perspective. PMID- 12576540 TI - Disasters and bioterrorism: does management training develop readiness? PMID- 12576543 TI - Matchmaking and species marriage: a game-theory model of community assembly. AB - Theories developed to explain the processes that govern the assembly and composition of natural plant communities can be divided into two broad categories. Niche-based theories propose that coevolutionary changes among species lead to character divergence (displacement), which allows for coexistence by partitioning resources among species. In contrast, ecological-drift theories propose that species diversity results from a balance of migration, speciation, and extinction, with little microevolutionary change. We use a game-theory model to reconcile drift and niche perspectives by developing a theory of species "marriage." Initially, ecological drift may determine which species encounter each other in a competitive arena. Once species come into contact, historical niche development as a result of prior coevolutionary molding of competitive ability determines which species may coexist. The model shows that only species that display the well-known tradeoff between seed size and competitive ability as a result of past competitive histories comply with the requisite for mutual evolutionary stability. Mutual evolutionary stability of competitive ability tends to make reproductive outputs more similar between species, increasing the chances of ecological equilibrium, i.e., the coexistence of species competing for a single resource. Moreover, mutual evolutionary stability guarantees that such an ecological equilibrium will be stable. The species-marriage model predicts that two or more plant species will coexist indefinitely (i.e., "marry") when their difference in seed size, their densities, and the resource availability obey a specific quantitative relation. For example, when resource availability is high, married species should be characterized by a greater asymmetry in seed size than when resource availability is low. Thus, in the species-marriage model, competition can shape the detailed properties of communities without violating the postulates of ecological-drift theory. PMID- 12576544 TI - Disruption of 3D tissue integrity facilitates adenovirus infection by deregulating the coxsackievirus and adenovirus receptor. AB - The human coxsackievirus and adenovirus receptor (CAR) represents the primary cellular site of adenovirus attachment during infection. An understanding of the mechanisms regulating its expression could contribute to improving efficacy and safety of adenovirus-based therapies. We characterized regulation of CAR expression in a 3D cell culture model of human breast cancer progression, which mimics aspects of the physiological tissue context in vitro. Phenotypically normal breast epithelial cells (S1) and their malignant derivative (T4-2 cells) were grown either on tissue culture plastic (2D) or 3D cultures in basement membrane matrix. S1 cells grown in 3D showed low levels of CAR, which was expressed mainly at cell-cell junctions. In contrast, T4-2 cells expressed high levels of CAR, which was mainly in the cytoplasm. When signaling through the epidermal growth factor receptor was inhibited in T4-2 cells, cells reverted to a normal phenotype, CAR protein expression was significantly reduced, and the protein relocalized to cell-cell junctions. Growth of S1 cells as 2D cultures or in 3D in collagen-I, a nonphysiological microenvironment for these cells, led to up-regulation of CAR to levels similar to those in T4-2 cells, independently of cellular growth rates. Thus, expression of CAR depends on the integrity and polarity of the 3D organization of epithelial cells. Disruption of this organization by changes in the microenvironment, including malignant transformation, leads to up-regulation of CAR, thus enhancing the cell's susceptibility to adenovirus infection. PMID- 12576545 TI - GDE1/MIR16 is a glycerophosphoinositol phosphodiesterase regulated by stimulation of G protein-coupled receptors. AB - Previously we identified MIR16 (membrane interacting protein of RGS16) as an integral membrane glycoprotein that interacts with regulator of G protein signaling proteins and shares significant sequence homology with bacterial glycerophosphodiester phosphodiesterases (GDEs), suggesting that it is a putative mammalian GDE. Here we show that MIR16 belongs to a large, evolutionarily conserved family of GDEs with a characteristic putative catalytic domain that shares a common motif (amino acids 92-116) with the catalytic domains of mammalian phosphoinositide phospholipases C. Expression of wild-type MIR16 (renamed GDE1), but not two catalytic domain mutants (E97A/D99A and H112A), leads to a dramatic increase in glycerophosphoinositol phosphodiesterase (GPI-PDE) activity in HEK 293T cells. Analysis of substrate specificity shows that GDE1/MIR16 selectively hydrolyzes GPI over glycerophosphocholine. The GPI-PDE activity of GDE1/MIR16 expressed in HEK 293T cells can be regulated by stimulation of G protein-coupled, alpha/beta-adrenergic, and lysophospholipid receptors. Membrane topology studies suggest a model in which the catalytic GDE domain faces the lumenextracellular space and the C terminus faces the cytoplasm. Our results suggest that by serving as a PDE for GPI with its activity regulated by G protein signaling, GDE1/MIR16 provides a link between phosphoinositide metabolism and G protein signal transduction. PMID- 12576546 TI - Contribution of transplanted bone marrow cells to Purkinje neurons in human adult brains. AB - We show here that cells within human adult bone marrow can contribute to cells in the adult human brain. Cerebellar tissues from female patients with hematologic malignancies, who had received chemotherapy, radiation, and a bone marrow transplant, were analyzed. Brain samples were obtained at autopsy from female patients who received male (sex-mismatched) or female (sex-matched, control) bone marrow transplants. Cerebella were evaluated in 10-microm-thick, formaldehyde fixed, paraffin-embedded sections that encompassed up to approximately 50% of a human Purkinje nucleus. A total of 5,860 Purkinje cells from sex-mismatched females and 3,202 Purkinje cells from sex-matched females were screened for Y chromosomes by epifluorescence. Confocal laser scanning microscopy allowed definitive identification of the sex chromosomes within the morphologically distinct Purkinje cells. In the brains of females who received male bone marrow, four Purkinje neurons were found that contained an X and a Y chromosome and two other Purkinje neurons contained more than a diploid number of sex chromosomes. No Y chromosomes were detected in the brains of sex-matched controls. The total frequency of male bone marrow contribution to female Purkinje cells approximated 0.1%. This study demonstrates that although during human development Purkinje neurons are no longer generated after birth, cells within the bone marrow can contribute to these CNS neurons even in adulthood. The underlying mechanism may be caused either by generation de novo of Purkinje neurons from bone marrow derived cells or by fusion of marrow-derived cells with existing recipient Purkinje neurons. PMID- 12576547 TI - Defective presentation of the CD1d1-restricted natural Va14Ja18 NKT lymphocyte antigen caused by beta-D-glucosylceramide synthase deficiency. AB - Va14Ja18 natural T (NKT) cells play an immunoregulatory role, which is controlled by a self glycolipid(s) presented by CD1d. Although the synthetic antigen alpha-D galactosylceramide (alpha-D-GalCer) stimulates all Va14Ja18 NKT cells, alpha anomeric D-glycosylceramides are currently unknown in mammals. We have used beta D-GalCer-deficient mice and beta-D-glucosylceramide (beta-D-GlcCer)-deficient cells to define the chemical nature of a natural NKT cell antigen. beta-D-GalCer deficient mice exhibit normal NKT cell development and function, and cells from these animals potently stimulate NKT hybridomas. In striking contrast, the same hybridomas fail to react to CD1d1 expressed by a beta-D-GlcCer-deficient cell line. Importantly, human beta-D-GlcCer synthase cDNA transfer, and hence the biosynthesis of beta-D-GlcCer, restores the recognition of mutant cells expressing CD1d1 by the Va14Ja18 NKT hybridomas. Additionally, suppression of beta-D-GlcCer synthesis inhibits antigen presentation to Va14Ja18 NKT cells. The possibility that beta-D-GlcCer itself is the natural NKT cell antigen was excluded because it was unable to activate NKT hybridomas in a cell-free antigen presentation assay. These findings suggest that beta-D-GlcCer may play an important role in generating and/or loading a natural Va14Ja18 NKT antigen. PMID- 12576548 TI - Evidence for the production of trioxygen species during antibody-catalyzed chemical modification of antigens. AB - Recent work in our laboratory showed that products formed by the antibody catalyzed water-oxidation pathway can kill bacteria. Dihydrogen peroxide, the end product of this pathway, was found to be necessary, but not sufficient, for the observed efficiency of bacterial killing. The search for further bactericidal agents that might be formed along the pathway led to the recognition of an oxidant that, in its interaction with chemical probes, showed the chemical signature of ozone. Here we report that the antibody-catalyzed water-oxidation process is capable of regioselectively converting antibody-bound benzoic acid into para-hydroxy benzoic acid as well as regioselectively hydroxylating the 4 position of the phenyl ring of a single tryptophan residue located in the antibody molecule. We view the occurrence of these highly selective chemical reactions as evidence for the formation of a short-lived hydroxylating radical species within the antibody molecule. In line with our previously presented hypothesis according to which the singlet-oxygen ((1)O*(2)) induced antibody catalyzed water-oxidation pathways proceeds via the formation of dihydrogen trioxide (H(2)O(3)), we now consider the possibility that the hydroxylating species might be the hydrotrioxy radical HO(3)*, and we point to the remarkable potential of this either H(2)O(3)- or O(3)-derivable species to act as a masked hydroxyl radical HO* in a biological environment. PMID- 12576549 TI - Suberoylanilide hydroxamic acid, a histone deacetylase inhibitor, ameliorates motor deficits in a mouse model of Huntington's disease. AB - Huntington's disease (HD) is an inherited, progressive neurological disorder that is caused by a CAG/polyglutamine repeat expansion and for which there is no effective therapy. Recent evidence indicates that transcriptional dysregulation may contribute to the molecular pathogenesis of this disease. Supporting this view, administration of histone deacetylase (HDAC) inhibitors has been shown to rescue lethality and photoreceptor neurodegeneration in a Drosophila model of polyglutamine disease. To further explore the therapeutic potential of HDAC inhibitors, we have conducted preclinical trials with suberoylanilide hydroxamic acid (SAHA), a potent HDAC inhibitor, in the R6/2 HD mouse model. We show that SAHA crosses the blood-brain barrier and increases histone acetylation in the brain. We found that SAHA could be administered orally in drinking water when complexed with cyclodextrins. SAHA dramatically improved the motor impairment in R6/2 mice, clearly validating the pursuit of this class of compounds as HD therapeutics. PMID- 12576550 TI - Communication between neocortex and hippocampus during sleep in rodents. AB - Both neocortical and hippocampal networks organize the firing patterns of their neurons by prominent oscillations during sleep, but the functional role of these rhythms is not well understood. Here, we show a robust correlation of neuronal discharges between the somatosensory cortex and hippocampus on both slow and fine time scales in the mouse and rat. Neuronal bursts in deep cortical layers, associated with sleep spindles and delta waves/slow rhythm, effectively triggered hippocampal discharges related to fast (ripple) oscillations. We hypothesize that oscillation-mediated temporal links coordinate specific information transfer between neocortical and hippocampal cell assemblies. Such a neocortical hippocampal interplay may be important for memory consolidation. PMID- 12576551 TI - Tissue transglutaminase-induced aggregation of alpha-synuclein: Implications for Lewy body formation in Parkinson's disease and dementia with Lewy bodies. AB - Proteinaceous aggregates containing alpha-synuclein represent a feature of neurodegenerative disorders such as Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. Despite extensive research, the mechanisms underlying alpha-synuclein aggregation remain elusive. Previously, tissue transglutaminase (tTGase) was found to contribute to the generation of aggregates by cross-linking pathogenic substrate proteins in Huntington's and Alzheimer's diseases. In this article, the role of tTGase in the formation of alpha-synuclein aggregates was investigated. Purified tTGase catalyzed alpha-synuclein cross linking, leading to the formation of high molecular weight aggregates in vitro, and overexpression of tTGase resulted in the formation of detergent-insoluble alpha-synuclein aggregates in cellular models. Immunocytochemical studies demonstrated the presence of alpha-synuclein-positive cytoplasmic inclusions in 8% of tTGase-expressing cells. The formation of these aggregates was significantly augmented by the calcium ionophore and prevented by the inhibitor cystamine. Immunohistochemical studies on postmortem brain tissue confirmed the presence of transglutaminase-catalyzed epsilon (gamma-glutamyl)lysine cross-links in the halo of Lewy bodies in Parkinson's disease and dementia with Lewy bodies, colocalizing with alpha-synuclein. These findings, taken together, suggest that tTGase activity leads to alpha-synuclein aggregation to form Lewy bodies and perhaps contributes to neurodegeneration. PMID- 12576553 TI - Negative specific heat of a magnetically self-confined plasma torus. AB - It is shown that the thermodynamic maximum-entropy principle predicts negative specific heat for a stationary, magnetically self-confined current-carrying plasma torus. Implications for the magnetic self-confinement of fusion plasma are considered. PMID- 12576552 TI - Multiple elements of the allergic arm of the immune response modulate autoimmune demyelination. AB - Analysis of mRNA from multiple sclerosis lesions revealed increased amounts of transcripts for several genes encoding molecules traditionally associated with allergic responses, including prostaglandin D synthase, histamine receptor type 1 (H1R), platelet activating factor receptor, Ig Fc epsilon receptor 1 (Fc epsilon RI), and tryptase. We now demonstrate that, in the animal model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE), mediated by T helper 1 (Th1) T cells, histamine receptor 1 and 2 (H1R and H2R) are present on inflammatory cells in brain lesions. Th1 cells reactive to myelin proteolipid protein expressed more H1R and less H2R than Th2 cells. Pyrilamine, an H1R antagonist, blocked EAE, and the platelet activating factor receptor antagonist CV6209 reduced the severity of EAE. EAE severity was also decreased in mice with disruption of the genes encoding Ig Fc gamma RIII or both Fc gamma RIII and Fc epsilon RI. Prostaglandin D synthase and tryptase transcripts were elevated in EAE brain. Taken together, these data reveal extensive involvement of elements of the immune response associated with allergy in autoimmune demyelination. The pathogenesis of demyelination must now be viewed as encompassing elements of both Th1 responses and "allergic" responses. PMID- 12576554 TI - Activated microglia in cortex of mouse models of mucopolysaccharidoses I and IIIB. AB - Alpha-N-acetylglucosaminidase deficiency (mucopolysaccharidosis IIIB, MPS IIIB) and alpha-l-iduronidase deficiency (MPS I) are heritable lysosomal storage diseases; neurodegeneration is prominent in MPS IIIB and in severe cases of MPS I. We have obtained morphologic and molecular evidence for the involvement of microglia in brain pathology of mouse models of the two diseases. In the cortex, a subset of microglia (sometimes perineuronal) consists of cells that are probably phagocytic; they have large storage vacuoles, react with MOMA-2 (monoclonal antibody against macrophages) and Griffonia simplicifolia isolectin IB(4), and stain intensely for the lysosomal proteins Lamp-1, Lamp-2, and cathepsin D as well as for G(M3) ganglioside. MOMA-2-positive cells appear at 1 and 6 months in MPS IIIB and MPS I mice, respectively, but though their number increases with age, they remain sparse. However, a profusion of cells carrying the macrophage CD68/macrosialin antigen appear in the cortex of both mouse models at 1 month. mRNA encoding CD68/macrosialin also increases at that time, as shown by microarray and Northern blot analyses. Ten other transcripts elevated in both mouse models are associated with macrophage functions, including complement C4, the three subunits of complement C1q, lysozyme M, cathepsins S and Z, cytochrome b558 small subunit, macrophage-specific protein 1, and DAP12. An increase in IFN gamma and IFN-gamma receptor was observed by immunohistochemistry. These functional increases may represent activation of resident microglia, an influx and activation of blood monocytes, or both. They show an inflammatory component of brain disease in the two MPS, as is known for many neurodegenerative disorders. PMID- 12576555 TI - Experimental test of a method for determining causal connectivities of species in reactions. AB - Theoretical analysis has shown the possibility of determining causal connectivities of reacting species and the reaction mechanism in complex chemical and biochemical reaction systems by applying pulse changes of concentrations of one or more species, of arbitrary magnitude, and measuring the temporal response of as many species as possible. This method, limited to measured and pulsed species, is given here an experimental test on a part of glycolysis including the sequence of reactions from glucose to fructose 1,6-biphosphate, followed by the bifurcation of that sequence into two branches, one ending in glycerol 3 phosphate, the other in glyceraldehyde 3-phosphate. Pulses of concentrations of one species at a time are applied to the open system in a non-equilibrium stationary state, and the temporal responses in concentrations of six metabolites are measured by capillary zone electrophoresis. From the results of these measurements and the use of the theory for their interpretation, we establish the causal connectivities of the metabolites and thus the reaction mechanism, including the bifurcation of one chain of reactions into two. In this test case of the pulse method, no prior knowledge was assumed of the biochemistry of this system. We conclude that the pulse method is relatively simple and effective in determining reaction mechanisms in complex systems, including reactants, products, intermediates, and catalysts and their effectors. The method is likely to be useful for substantially more complex systems. PMID- 12576557 TI - Health assessment in aviation medicine: an overview. PMID- 12576558 TI - Fitness standards in airline staff. AB - Employers have a legal duty to manage safety and to meet this duty it is necessary to control risks arising from pre-existing medical conditions. Such conditions can be identified at the time of job placement, but there is little published information about the approach to fitness standards by most airlines. This review is therefore limited to the practice of one major airline. It is illustrated by reference to various occupations within the airline, with the associated rationale. PMID- 12576559 TI - Aircrew periodic medical examinations. AB - BACKGROUND: Aircrew are subject to periodic medical examinations (PMEs) throughout their careers. These aim to ensure flight safety. AIMS: This paper discusses the history and rationale of screening aircrew. It then considers the ability of the PME to detect abnormalities and predict future in-flight incapacitation. CONCLUSION: We conclude that the current system is of limited value. Selected changes to the content and frequency of aircrew periodic medical examinations could be made without compromising flight safety. PMID- 12576560 TI - The value of screening tests in applicants for professional pilot medical certification. AB - Good medicine does not consist in the indiscriminate application of laboratory examinations to a patient, but rather in having so clear a comprehension of the probabilities and possibilities of a case as to know what tests may be expected to give information of value.' (Francis Peabody, 1922) PMID- 12576561 TI - Customer health: a new role for occupational physicians. AB - BACKGROUND: Many employees are required to travel by air as part of their work, and occupational physicians may be asked to give travel health advice and to carry out risk assessments. AIMS: This review examines the stresses of air travel, including those due to travel to and transit through the airport, as well as those arising from the aircraft cabin environment. Guidance is given on the potential effects of these stressors, particularly on those with certain underlying medical conditions, and advice given on how to mitigate some of the effects. Finally, the review looks at in-flight medical incidents and recent developments in in-flight medical care. CONCLUSION: Despite the stresses of air travel, most people are able to fly safely. Some individuals are more vulnerable to these stresses and occupational physicians have an important role in ensuring that an appropriate risk assessment is carried out. PMID- 12576562 TI - Lung and bladder cancer among Danish urban bus drivers and tramway employees: a nested case-control study. AB - BACKGROUND: The combustion of fossil fuels produces small amounts of mutagenic and carcinogenic compounds. We investigated the association between employment and lung and bladder cancer in Danish bus drivers and tramway employees. METHODS: We carried out a nested case-control study of 153 lung and 84 bladder cancer cases, and 606 controls sampled in a cohort of 18 174 bus drivers or tramway employees employed in Copenhagen during the period 1900-1994. The cases and controls or their next of kin were interviewed about smoking, along with occupational and residential history. An exposure index based on which bus routes the bus drivers had mainly been driving was established. Relative risks (RRs) were estimated by conditional logistic regression. RESULTS: The analysis showed decreasing risk for lung cancer with increasing years of employment as a bus driver (RR = 0.97 for each added year, 95% confidence interval = 0.96-0.99). The air pollution index based on main bus for the bus drivers showed no positive correlation with risk. PMID- 12576563 TI - Microorganism-induced skin disease in workers exposed to metalworking fluids. AB - BACKGROUND: An outbreak of skin disorders among workers potentially exposed to metalworking fluids prompted the present study. Few studies have described skin disorders associated with microbe-contaminated metalworking fluids. METHODS: Samples of materials contaminated with metalworking fluids were obtained from two manufacturing facilities in Ohio. Pathogenic bacteria and yeasts, in concentrations sufficient to cause skin disease, were cultured from 9 of 12 (75%) sampled materials. RESULTS: Allergic patch testing of five affected people produced negative results for standard allergens, augmented by fluids and items from their workplace. This ruled out allergies as the cause of the skin disease. Improper handling and disposal of cotton gloves, inappropriate use of scouring pads and ineffective hand wiping were apparently responsible for the microbiological contamination. The hands and forearms were most commonly affected. CONCLUSION: Improper handling of soluble, synthetic and semi-synthetic metalworking fluids provides an excellent environment for the growth of a range of microorganisms, including bacteria and fungi. If allowed to grow because of poor occupational hygiene, these microorganisms can cause skin disorders among workers. Soiled protective clothing (gloves, coveralls and work boots) should be cleaned or discarded on a regular basis. When washing up, workers should not use metalworking fluids and items used to clean machinery. PMID- 12576564 TI - A cross-sectional study of employer and employee occupational health needs and priorities within the Irish Civil Service. AB - BACKGROUND: This study aimed to establish how important an occupational health unit (OHU) is to its clients, and to identify the perceived needs and priorities for such a service. METHODS: A cross-sectional postal survey of a stratified, randomly selected group of employees (n = 760) and all human resources (HR) managers (n = 34) was conducted in the Irish Civil Service. Each participant was requested to rate the overall importance of the OHU and to prioritize eight proposed functions for the unit: medical surveillance, general health education, pre-employment/promotion medical assessments, ill-health retirement assessments, return-to-work (after sick leave) assessments, occupational health education, research, and general medical screening. The results were analysed according to age group, gender, grade and occupation. RESULTS: There was a response rate of 69% from employees and 74% from personnel managers. Significantly more HR managers than employees (92 versus 81%) thought an occupational health service was either important or very important. There were also differences in prioritization of functions by employees and HR managers. HR managers prioritized those functions concerned with assessing individuals' fitness for work, notably pre-employment/promotional health assessments, whereas employees consider group directed 'preventative' functions to be more important, i.e. general medical screening, health education and medical surveillance. Both sets of opinions are not mutually exclusive, and considerable overlap exists, notably in the areas of occupational and general health education. PMID- 12576565 TI - NHS occupational health services in England and Wales--a changing picture. AB - AIMS: To establish the extent of doctor input to occupational health (OH) service provision in the UK National Health Service (NHS) in 2001 and to compare this with inputs in 1998. METHOD: A postal questionnaire was used to obtain information from OH medical staff employed by the NHS in England and Wales. RESULTS: The NHS OH service has seen an increase between 1998 and 2001 in the amount of doctor time per employee. Doctors tend to work now for more sessions per week. The proportion of doctors holding specialist qualifications has also increased. An increased number of NHS employees now have access to consultant care for occupational medicine. OH departments increasingly tend to provide services to employees beyond the NHS and are thereby able to generate income to further the development of the service. CONCLUSIONS: Steady progress is being made in improving the provision of OH services within the NHS. However, substantial variation exists in the apparent level of access to such provision. The government policy for all NHS staff to have access to a consultant-led service is not yet met. NHS Plus will impact on this picture and deserves study in the future. PMID- 12576566 TI - The effect on attendance rates for sickness absence assessment of a revised employee information sheet. AB - BACKGROUND: An increasingly high standard of ethical practice is expected of all doctors and in particular those not providing treatment services. AIMS: This case control study investigated the effect on non-attendance rates for first sickness absence appointments of a new employee information sheet sent to staff from the two largest departments, education and social services, of a large UK local authority. METHOD: An information sheet detailing the ethical standards applying to a local authority occupational health service was developed and sent to all employees referred by their managers as part of the sickness absence management procedures, along with details of their appointment. RESULTS: A reduction of approximately 1/3 in the rate of non-attendance was noted in the intervention group, which was statistically significant for social services referrals. CONCLUSION: This approach may improve the efficiency and effectiveness of occupational health services, as well as helping to achieve the informed consent of employees undergoing occupational health assessment. PMID- 12576567 TI - Interview data versus questionnaire data in the diagnosis of carpal tunnel syndrome in epidemiological studies. AB - AIMS: In most epidemiological studies on carpal tunnel syndrome (CTS), the case definition is based on questionnaire data with or without neurophysiological testing. The aim of this study was to test if the use of questionnaire data results in misclassification of cases. METHODS: In two studies, involving 940 and 311 participants, respectively, the people indicating CTS symptoms (tingling) in a questionnaire were clinically interviewed. In a subpopulation (n = 404), all went through an interview regardless of their questionnaire answers. RESULTS: Only 35-45% of the participants reporting tingling once a week or more in the questionnaire actually had symptoms consistent with CTS when interviewed. The remaining 55-65% had no or infrequent symptoms or symptoms because of other disorders. Few potential CTS cases were missed. The positive predictive values were 0.48 (95% CI = 0.30-0.66) and 0.52 (95% CI = 0.38-0.67) on the right and left hands, respectively. The sensitivities, specificities and negative predictive values ranged from 0.87 to 1.00. CONCLUSIONS: Questionnaire information overestimates the prevalence of CTS symptoms. However, asking about tingling in a questionnaire is a simple and sensitive first step to detect potential CTS cases, but symptoms should be confirmed by interview. PMID- 12576568 TI - Factors associated with premature departure from working life among ageing food industry employees. AB - AIM: To find out how health status, work ability and job-related factors were associated with premature departure from working life in an 11 year follow-up period. METHODS: The study population was composed of 126 ageing food industry employees. Baseline data were derived from medical examinations and self completed questionnaires in 1989. The follow-up data were collected in 2000. Data analyses were performed by independent samples t-tests and Mann-Whitney and chi(2) tests. RESULTS: Several chronic diseases, higher degree of work impairment due to diseases, long-term sickness absence, stress symptoms, low scores in work ability index, poor subjective work ability and physical work ability, and heavy physical workload seemed to be factors associated with early departure from working life among ageing food industry employees. CONCLUSION: Identification of these factors, prevention of sickness and promotion of work ability seem to be the most important tools to reduce premature exit from working life. PMID- 12576570 TI - Dermal exposure assessment in occupational medicine. PMID- 12576574 TI - Prokaryote taxonomy of the 20th century and the impact of studies on the genus Pseudomonas: a personal view. AB - The taxonomic studies on the genus Pseudomonas performed in the Department of Bacteriology of the University of California at Berkeley played a significant role in the development of modern prokaryote taxonomy, which started in the 1960s. This impact was due to a revival of the method of den Dooren de Jong for the nutritional analysis of chemoorganotrophic organisms and, mostly, to the introduction of the determination of rRNA as a method of taxonomic analysis. While the introduction of the nutritional studies facilitated the characterization of Pseudomonas and other chemoorganotrophs, the applicability of the rRNA studies extended to all prokaryotes. PMID- 12576575 TI - Controlling competence in Bacillus subtilis: shared use of regulators. AB - Bacteria have developed a wide arsenal of survival strategies to cope with the specific problems posed by their environment. These processes are carefully regulated and complex signal transduction cascades ensure proper activation of the adequate adaptive response. An intriguing observation is that generally the regulation pathways of the different adaptive processes are highly intertwined. In this review, this phenomenon is illustrated by the regulation of genetic competence development in Bacillus subtilis. The different regulation pathways which make up the gene regulation network that controls the development of competence are described, and their connections to other adaptive processes in B. subtilis are discussed. PMID- 12576576 TI - A plasmid-borne Rap-Phr system of Bacillus subtilis can mediate cell-density controlled production of extracellular proteases. AB - Bacillus subtilis uses two-component signal transduction systems to sense intra- and extracellular stimuli to adapt to fluctuating environmental situations. Regulator aspartate phosphatases (Raps) have important roles in these processes, as they can dephosphorylate certain response-regulators, and are themselves subject to cell-density-controlled inhibition by secreted Phr (phosphate regulator) peptides. Eleven chromosomal genes encode this family of phosphatases, but in addition, certain strains contain endogenous plasmids with genes for homologous Rap-Phr systems. Plasmid pTA1060 encodes Rap60 and its antagonistic signalling molecule Phr60. Strikingly, expression of Rap60 in B. subtilis 168 strongly repressed the production of proteolytic enzymes. In fact, the transcription of the aprE gene, encoding a major extracellular protease, was shown to be decreased upon Rap60 expression, whereas this effect could be antagonized by the extracellular addition of synthetic Phr60 pentapeptide. Finally, transcription studies suggest that Rap60 dephosphorylates a component of the phosphorelay and is coupled to aprE transcription by the transition-state regulator AbrB. In conclusion, these data show that endogenous plasmids contain functional Rap-Phr systems and for the first time, that Rap-Phr systems can mediate cell-density controlled production of secreted proteases. This quorum sensing mechanism might enable B. subtilis to suppress protease production under conditions of low cell densities when nutrients are still available in sufficient amounts. PMID- 12576577 TI - Transcriptional regulation of the nos genes for nitrous oxide reductase in Pseudomonas aeruginosa. AB - The genes for nitrous oxide (N(2)O) reduction, nosRZDFYL, are clustered on the chromosome of Pseudomonas aeruginosa. Promoter assays using transcriptional fusions to lacZ revealed that the structural gene for nitrous oxide reductase, nosZ, is transcribed with the upstream nosR gene. The nosR gene product is not required for the activity of the nosR promoter. A sequence similar to the consensus FNR-binding motif was found 41.5 bp upstream from the major transcriptional start point of nosR. Mutation of the motif significantly reduced the promoter activity. DNR, an FNR-related transcriptional regulator required for the expression of denitrification genes in P. aeruginosa, is necessary for the transcription of nosR, indicating that the motif is recognized by DNR. Nitrite (NO-2), nitric oxide (NO) and NO-generating reagents induced nosR promoter activity, but N(2)O did not. The NO-2-induced nosR promoter activity was reduced by mutation of the NO-2 reductase gene. However, a low concentration of NO-2 induced the promoter activity in a NO reductase mutant. These results indicate that NO is the inducer molecule for transcription of the nos genes. PMID- 12576579 TI - Persistence of biofilm-associated Escherichia coli and Pseudomonas aeruginosa in groundwater and treated effluent in a laboratory model system. AB - This study was based on the hypothesis that groundwater-derived biofilms may provide a reservoir for coliform or pathogenic bacteria as has been observed in drinking water distribution systems. Escherichia coli, labelled with green fluorescent protein, was found to colonize all layers of mixed-population biofilms developed in association with indigenous groundwater micro-organisms in a laboratory-scale reactor. Biofilm-associated E. coli was removed at a slower rate from the reactor flasks than planktonic E. coli under a continuous flow regime. During flow-through of groundwater, planktonic E. coli removal was slower in flasks containing coverslips for enhanced biofilm development compared to a control flask without coverslips. Conversely, during flow-through of treated effluent, planktonic E. coli removal was faster in flasks with coverslips compared to without. Removal of attached E. coli was also fastest in the coverslip-containing flasks with effluent flow-through. This suggests that an increase in available nutrients may reduce E. coli survival potential due to either enhanced competition for nutrients or enhanced antagonism by the indigenous microbial population. Under identical conditions, GFP-labelled Pseudomonas aeruginosa was found to persist in the biofilms for longer than E. coli, most notably when exposed to flow-through of treated effluent. However, prolonged persistence of P. aeruginosa in the effluent could not be attributed to an association with the biofilms. This study has shown that under certain conditions the presence of mixed-population biofilms may limit the survival potential of enteric bacterial pathogens introduced into groundwater. PMID- 12576578 TI - Surface motility in Pseudomonas sp. DSS73 is required for efficient biological containment of the root-pathogenic microfungi Rhizoctonia solani and Pythium ultimum. AB - Pseudomonas sp. DSS73 was isolated from the rhizoplane of sugar beet seedlings. This strain exhibits antagonism towards the root-pathogenic microfungi Pythium ultimum and Rhizoctonia solani. Production of the cyclic lipopeptide amphisin in combination with expression of flagella enables the growing bacterial culture to move readily over the surface of laboratory media. Amphisin is a new member of a group of dual-functioning compounds such as tensin, viscosin and viscosinamid that display both biosurfactant and antifungal properties. The ability of DSS73 to efficiently contain root-pathogenic microfungi is shown to arise from amphisin dependent surface translocation and growth by which the bacterium can lay siege to the fungi. The synergistic effects of surface motility and synthesis of a battery of antifungal compounds efficiently contain and terminate growth of the microfungi. PMID- 12576580 TI - Diet influences the ecology of lactic acid bacteria and Escherichia coli along the digestive tract of cattle: neural networks and 16S rDNA. AB - In this manuscript, the authors have sought to gain a better understanding of the interactions between Escherichia coli and lactic acid bacteria (LAB) isolated from Rogossa MRS agar along the digestive tract of grain- and forage-fed cattle. E. coli from cattle receiving a high-grain diet were more numerous (P<0.05) than from the high-forage diet and the highest numbers were in the faeces. Isolates on Rogossa MRS agar were always greater in the high-grain diet (P<0.05) and contained a significant number of LAB. A random set of Rogossa MRS agar colonies was selected and artificial neural networks were used to develop a relationship between colony description and species which was validated using sequence analysis (16S rDNA). The neural networks correctly predicted species in more than 80 % of cases and was composed, primarily, of Lactobacillus vitulinus, Lactobacillus ruminis, Selenomonas ruminantium, Streptococcus bovis, Acidaminococcus fermentans and Megasphaera elsdenii. In conjunction with statistical diversity indices, it was demonstrated that diversity in the high fibre diet was always lower and was a consequence of the dominance of Str. bovis. In contrast, the diversity in the high-grain diet was greater (P<0.05) and was a consequence of the decline in Str. bovis. These data demonstrate that there is a positive relationship between coliform and LAB isolates throughout the digestive tract of cattle, and diet is the major factor regulating bacterial composition. PMID- 12576581 TI - Molecular analysis of bacteria in periodontitis: evaluation of clone libraries, novel phylotypes and putative pathogens. AB - Subgingival plaque samples were obtained from 26 subjects with advanced periodontal lesions. Bacterial diversity was analysed by amplification of the 16S rRNA genes with two different primer sets, and subsequent cloning and sequencing. A total of 578 sequences was analysed after the exclusion of chimeras. The authors found 148 phylotypes with the clone library 27f/519r (number of clones n=322; coverage, C=66 %) and 75 phylotypes with the clone library 515f/1525r (n=256; C=84 %). Comparative sequence analysis revealed that 70 % of all of the analysed sequences showed a similarity of at least 99 % to sequences deposited in public databases. The classes Actinobacteria, Bacilli, Bacteroidetes, Clostridia, Deferribacteres, Flavobacteria, Fusobacteria, Mollicutes, Spirochaetes and all classes of the Proteobacteria were represented. Sequences that were at least 99 % identical to Porphyromonas gingivalis, Filifactor alocis and Treponema socranskii were present in at least one-third of the patients. Libraries generated with the two PCR primer pairs differed significantly in their representation of the families Porphyromonadaceae, Prevotellaceae, Fusobacteriaceae, Eubacteriaceae, Streptococcaceae and ACIDAMINOCOCCACEAE: A total of 14 sequences exhibited less than 97 % identity to sequences published previously and were assigned to six different families within the phyla Bacteroidetes and FIRMICUTES: Twelve of 20 putative pathogens were recovered, which were recently proposed to be associated with periodontitis. PMID- 12576582 TI - Population structure analysis of Burkholderia cepacia genomovar III: varying degrees of genetic recombination characterize major clonal complexes. AB - Infection with bacterial species belonging to the Burkholderia cepacia complex contribute significantly to morbidity and mortality in persons with cystic fibrosis (CF). The majority of isolates recovered from CF patients belong to B. cepacia genomovar III and several distinct 'epidemic' strains have been described. This study examined the population structure of B. cepacia genomovar III by using multilocus restriction typing, indexing allelic variation at five chromosomal genes by restriction analysis of PCR-amplified genes. A collection of 375 isolates, recovered from CF and non-CF patients and natural environments in North America, Europe and Australia, was examined. Among these isolates 144 different restriction types were found. Overall, the population is at linkage disequilibrium, indicating that it has a clonal structure. The majority (86.7 %) of restriction types grouped into three major clonal complexes, comprising the epidemic ET12, PHDC and Midwest clonal lineages. The analysis indicates that these complexes are geographically widespread and demonstrate varying degrees of genetic recombination. These differences in population structure among major clonal complexes within the same species are likely related to differences in evolutionary history and ecology. The observation that genetic recombination is frequent within some B. cepacia genomovar III populations has important implications for the biotechnological use of B. cepacia complex species. PMID- 12576583 TI - Genesis of variants of Vibrio cholerae O1 biotype El Tor: role of the CTXphi array and its position in the genome. AB - The gene encoding cholera toxin, the principal virulence factor of Vibrio cholerae, is encoded by a filamentous, lysogenic bacteriophage known as CTXphi. The genome of V. cholerae, the host for CTXphi, consists of two chromosomes, one large and one small. Here, it is shown that localization and array of CTX prophage DNA in either the large or small chromosome of V. cholerae is likely to be one of the reasons for the emergence of O1 biotype El Tor variants isolated just before and after the V. cholerae O139 cholera outbreak in 1992. Analyses of the organization of the CTX region of the genome of pre-O139 El Tor strains revealed that these strains carry two distinct CTX prophages integrated in the small chromosome in tandem: CTX(ET), the prophage having a conserved NotI site in its repeat sequence segment which seems to be specific for the El Tor strains so far examined, followed by CTX(calc)-like genome, the prophage found in recent O139 clinical isolates from Calcutta. In sharp contrast, in post-O139 El Tor strains only one copy of the CTX(ET) prophage was found to be integrated in the large chromosome. To the authors' knowledge, the presence of CTX prophage in the small chromosome of O1 El Tor strains has not been reported previously. It is also shown that the difference in the CTX copy number and the position of the bacteriophage on the genomes of pre- and post-O139 El Tor strains have an effect on cholera toxin production. While a pre-O139 strain produced maximum cholera toxin in yeast extract/peptone medium at 30 degrees C, a post-O139 El Tor strain showed maximal yield at 37 degrees C, indicating differential regulation of cholera toxin between the strains. It appears from this study that the variation in the integration site of the CTX prophage, its copy number and the presence of diverse phage genomes in V. cholerae O1 biotype El Tor may be strategically important for generating variants with subtle phenotypic modulations of virulence factor production in this longest-ruling seventh pandemic strain. PMID- 12576584 TI - Analysis of mutations in the Staphylococcus aureus clfB promoter leading to increased expression. AB - A clfB : : tetK reporter was constructed in Staphylococcus aureus strains Newman and 8325-4, whereby the level of tetracycline resistance reflected the activity of the clfB promoter. Wild-type strains carrying a single copy of this construct exhibited a low level of tetracycline resistance, suggesting that the clfB promoter is weak. Spontaneous mutants that grew at higher tetracycline concentrations were isolated. Some were due to point mutations in the clfB promoter that led to increased expression of the tetK gene. The clfB promoter was identified by primer extension analysis and -35 and -10 elements were assigned. The promoter regions from the tetracycline-resistant mutants were sequenced and several had base changes within or adjacent to the -35 box. Three created the consensus -35 sequence TTGACA. The mutant clfB promoters were fused to lacZ. beta Galactosidase activity was six- to ninefold higher in the mutant strains compared to the wild-type. The wild-type clfB gene was placed under the control of the mutant promoters. ClfB expression was higher than the corresponding wild-type strains and the protein was present on bacteria from the stationary phase instead of being confined to the exponential phase. Therefore, mutations in the clfB promoter that cause changes in the -35 region produce a stronger promoter that is capable of increased transcription and, as a result, increased expression of ClfB. PMID- 12576585 TI - Negative regulation of PrfA, the key activator of Listeria monocytogenes virulence gene expression, is dispensable for bacterial pathogenesis. AB - Listeria monocytogenes is a facultative intracellular bacterial pathogen that regulates the expression of virulence-associated gene products in response to specific host cell compartment environments. The PrfA protein of L. monocytogenes functions as a key regulatory factor required for the differential expression of bacterial virulence gene products within infected host cells. PrfA both positively and negatively regulates its own expression, and while PrfA positive regulation is required for cell-to-cell spread of L. monocytogenes and for full virulence in infected mice, a role for negative regulation has been of presumed importance but has yet to be established. To address the role of negative regulation of prfA expression in L. monocytogenes pathogenesis, prfA promoter mutations designed to reduce or eliminate negative regulation were introduced into L. monocytogenes and analysed for their effects on patterns of PrfA dependent gene expression and virulence in murine models of infection. High level PrfA production resulting from the prfA promoter mutations produced significantly increased levels of PrfA-regulated gene expression in broth-grown cultures; however the apparent loss of negative prfA regulation had no deleterious effects on growth and spread of the bacteria within infected tissue culture cells or on virulence in mice. The results indicate that while negative regulation of prfA expression exists and provides a feedback system for the control of PrfA synthesis, this feedback system is dispensable for virulence. PMID- 12576586 TI - Role of the thioredoxin system and the thiol-peroxidases Tpx and Bcp in mediating resistance to oxidative and nitrosative stress in Helicobacter pylori. AB - Helicobacter pylori possesses two distinct thioredoxin proteins (Trx1 and Trx2) which may play important roles in the ability of this bacterium to survive oxidative stress. Trx1 has previously been shown to be an electron donor in vitro for alkyl-hydroperoxide reductase (AhpC), one of three members of the peroxiredoxin family of antioxidant peroxidases present in H. pylori. In this study, mutants in the trxA1 and trxA2 genes encoding Trx1 and Trx2, respectively, and in the tpx and bcp genes, which encode the remaining two members of the H. pylori peroxiredoxin family, were constructed in order to determine their roles in resistance to damage by reactive oxygen and nitrogen species. Mutation of trxA1 led to a pronounced increase in sensitivity to oxygen, hydrogen peroxide and the superoxide generator paraquat, as well as to the nitric oxide (NO) releasers sodium nitroprusside (SNP) and S-nitrosoglutathione (GSNO), consistent with an in vivo role for Trx1 as a reductant for AhpC. A trxA2 single mutant grew normally in an atmosphere of 2 % (v/v) O(2) but grew very poorly in 10 % (v/v) O(2). It showed slight increases in killing by hydrogen peroxide, paraquat, SNP and GSNO compared to the wild-type, but was significantly more sensitive to cumene hydroperoxide in disc-diffusion assays. A trxA1 trxA2 double mutant was very sensitive to all of the oxidative and nitrosative stresses applied. Comparison of the phenotypes of the tpx and bcp mutants showed that Tpx plays a significant role in peroxide and superoxide resistance in H. pylori, while the role of Bcp is minimal. No evidence was obtained for a role for either Tpx or Bcp in resistance to the toxic effects of NO. The results show that a functional thioredoxin system is necessary for both oxidative and nitrosative stress resistance in H. pylori but, surprisingly, is not essential for viability despite the absence of glutathione and a glutaredoxin system in this bacterium. PMID- 12576587 TI - Molecular and genetic characterization of a serotype A MATa Cryptococcus neoformans isolate. AB - Cryptococcus neoformans comprises two varieties (neoformans and gattii) and four serotypes (A, B, C and D). Fertile isolates of both mating types have been identified in serotypes B, C and D; however, a fertile serotype A MATa strain has not been confirmed, although serotype A MATalpha strains will mate with serotype D MATa strains. Preliminary analysis of a recent Italian environmental isolate (IUM 96-2828) suggested that this strain was haploid, serotype A and MATa. In this study, IUM 96-2828 has been characterized in detail. A mating reaction between IUM 96-2828 and H99 (serotype A MATalpha) produced abundant spores with an equal distribution of MATa and MATalpha progeny, all of which were serotype A. Karyotypic analysis of F(1) spores revealed evidence of recombination, confirming that IUM 96-2828 was fertile. The MATa pheromone gene from IUM 96-2828 was sequenced and found to be most closely related to the serotype D MATa pheromone gene. Phylogenetic comparisons of other genes not linked to mating type also suggested IUM 96-2828 was most closely related to serotype A strains. Biochemical analysis showed that the carbon assimilation profiles of H99 and IUM 96-2828 were identical for 97 % (30/31) of the substrates while isozyme analysis showed 89 % (17/19) identity. Assays of major virulence factors found no difference between H99 and IUM 96-2828. Virulence studies using the mouse model demonstrated that IUM 96-2828 was virulent for mice, although it was less virulent than H99. These data strongly suggest that IUM 96-2828 is a true haploid serotype A MATa isolate that is fertile. PMID- 12576588 TI - Molecular analysis of Mycobacterium tuberculosis DNA from a family of 18th century Hungarians. AB - The naturally mummified remains of a mother and two daughters found in an 18th century Hungarian crypt were analysed, using multiple molecular genetic techniques to examine the epidemiology and evolution of tuberculosis. DNA was amplified from a number of targets on the Mycobacterium tuberculosis genome, including DNA from IS6110, gyrA, katG codon 463, oxyR, dnaA-dnaN, mtp40, plcD and the direct repeat (DR) region. The strains present in the mummified remains were identified as M. tuberculosis and not Mycobacterium bovis, from katG and gyrA genotyping, PCR from the oxyR and mtp40 loci, and spoligotyping. Spoligotyping divided the samples into two strain types, and screening for a deletion in the MT1801-plcD region initially divided the strains into three types. Further investigation showed, however, that an apparent deletion was due to poor DNA preservation. By comparing the effect of PCR target size on the yield of amplicon, a clear difference was shown between 18th century and modern M. tuberculosis DNA. A two-centre system was used to confirm the findings of this study, which clearly demonstrate the value of using molecular genetic techniques to study historical cases of tuberculosis and the care required in drawing conclusions. The genotyping and spoligotyping results are consistent with the most recent theory of the evolution and spread of the modern tuberculosis epidemic. PMID- 12576590 TI - Application of a novel multi-screening signature-tagged mutagenesis assay for identification of Klebsiella pneumoniae genes essential in colonization and infection. AB - Klebsiella pneumoniae is a common cause of urinary tract infections (UTIs) and pneumonia, especially in immunocompromised individuals. Epidemiological studies have revealed that K. pneumoniae infections are frequently preceded by gastrointestinal colonization and the gastrointestinal tract is believed to be the most important reservoir for transmission of the bacteria. To identify genes involved in the ability of K. pneumoniae to colonize the intestine and infect the urinary tract, a novel multi-screening signature-tagged mutagenesis (MS-STM) assay was implemented. In the MS-STM assay, PCR-amplified tags present in the inoculum as well as recovered pools from each infection model are simultaneously subjected to hybridization using each specific tag as a probe. Therefore, screenings of a mutant library in more than one infection model is significantly eased compared to the traditional signature-tagged mutagenesis methodology. From a total of 1,440 K. pneumoniae transposon mutants screened, 13 mutants were identified as attenuated in intestinal colonization as well as the UTI model. In addition, six mutants attenuated only in the UTI model were identified. Transposon insertion sites in attenuated mutants were, among others, in genes encoding well-known K. pneumoniae virulence factors such as lipopolysaccharide and capsule, as well as in genes of unknown function. PMID- 12576589 TI - Maximal adherence and invasion of INT 407 cells by Campylobacter jejuni requires the CadF outer-membrane protein and microfilament reorganization. AB - The binding of Campylobacter jejuni to fibronectin (Fn), a component of the extracellular matrix, is mediated by a 37 kDa outer-membrane protein termed CadF for Campylobacter adhesion to fibronectin. The specificity of C. jejuni binding to Fn, via CadF, was demonstrated using antibodies reactive against Fn and CadF. More specifically, the anti-CadF antibody reduced the binding of two C. jejuni clinical isolates to immobilized Fn by greater than 50 %. Furthermore, a C. jejuni wild-type isolate, in contrast to the isogenic CadF mutant, was found to compete with another C. jejuni wild-type isolate for host cell receptors. Given the relationship between the pericellular Fn matrix and the cytoskeleton, the involvement of host cell cytoskeletal components in C. jejuni internalization was also examined. Cytochalasin D and mycalolide B microfilament depolymerizing agents resulted in a significant reduction in C. jejuni invasion. Studies targeting paxillin, a focal adhesion signalling molecule, identified an increased level of tyrosine phosphorylation upon C. jejuni infection of INT 407 cells. Collectively, these data suggest CadF promotes the binding of C. jejuni to Fn, which in turn stimulates a signal transduction pathway involving paxillin. PMID- 12576591 TI - A novel ColV plasmid encoding type IV pili. AB - Many septicaemic Escherichia coli strains harbour ColV virulence plasmids. This paper describes pO78V, a conjugative ColV plasmid from an avian pathogenic E. coli strain that encodes type IV pili in addition to other virulence-related genes and tetracycline resistance. Plasmid location of type IV pili genes was demonstrated using Southern hybridization and expression of the pili was demonstrated using RT-PCR and phage sensitivity assays. This is a first report of a ColV plasmid encoding type IV pili. Plasmid pO78V is a mosaic plasmid containing replicons and other genes typical to both IncI1 and IncFII groups. As type IV pili of Gram-negative bacteria are involved in several stages of infection, their presence on a ColV virulence plasmid could expand the repertoire of pathogenesis-related genes. PMID- 12576592 TI - Characterization of LppS, an adhesin of Mycoplasma conjunctivae. AB - A serine-rich membrane protein named LppS from Mycoplasma conjunctivae, the aetiological agent of infectious keratoconjunctivitis (IKC) of domestic and wild Caprinae, was characterized. Gene cloning and sequence analysis of the lppS gene revealed that it encoded a membrane protein precursor. The protein had a typical signal sequence and a signal peptidase II cleavage site followed by a cysteine residue representing a potential acylation site. The mature LppS protein had an apparent molecular mass of 150 kDa and was found in the detergent-associated fraction of Tween 20 extracted M. conjunctivae proteins. It possessed a serine rich domain of 41 aa with 37 (90.2 %) serine residues. Twenty-seven of these serine residues were contiguous. The protein adhered to lamb joint synovial cells. Using an in vitro adhesion model, Fab fragments from IgG directed against recombinant purified LppS were shown to specifically inhibit adhesion of M. conjunctivae to lamb cells. Thus, LppS is likely to be an adhesin of M. conjunctivae that may play an important role in the pathogenesis of IKC. PMID- 12576593 TI - Characterization of a chromosomal region of Mycoplasma sp. bovine group 7 strain PG50 encoding a glycerol transport locus (gtsABC). AB - Mycoplasma species bovine group 7, represented by the type strain PG50, is one of six members of the Mycoplasma mycoides cluster and has been implicated in sporadic and outbreak cases of polyarthritis and mastitis in Australian dairy cattle. This study describes cloning and sequencing a 7.9 kb region of the PG50 chromosome and identification of genes involved in glycerol transport (gtsA, gtsB and gtsC) that are followed by a putative lipoprotein gene lppB and a genomic locus containing two ORFs encoding putative membrane proteins. Long range PCR using primers spanning gtsABC and downstream flanking genes, and Southern hybridization analyses using a suite of probes derived from M. mycoides subsp. mycoides small colony type (SC) strain Afade for gtsA, gtsB and gtsC, lppB and the two downstream genes confirmed that these genes were conserved among Mycoplasma sp. bovine group 7 isolates and mycoplasmas belonging to the M. mycoides subcluster [M. mycoides subsp. mycoides SC, M. mycoides subsp. mycoides large colony type (LC) and M. mycoides subsp. capri] but were absent in mycoplasmas belonging to the Mycoplasma capricolum subcluster (M. capricolum subsp. capricolum and M. capricolum subsp. capripneumoniae). M. capricolum subsp. capricolum type strain California kid did not hybridize with the probe for gtsA and gave only weak or no hybridization signals with probes derived from the loci downstream of gtsABC, suggesting that this region has diverged in mycoplasmas belonging to subspecies of M. capricolum. It is shown that PG50, after the addition of a physiological concentration of glycerol to the growth medium, generates H(2)O(2) at levels comparable with strain Afade, implying that the glycerol transport system is functional in Mycoplasma sp. bovine group 7. This suggests that in PG50, as in M. mycoides subsp. mycoides SC, glycerol uptake is followed by phosphorylation to glycerol 3-phosphate and then conversion to dihydroxyacetone phosphate, catalysed by L-alpha-glycerophosphate oxidase, resulting in the production of H(2)O(2). The ability of Mycoplasma sp. bovine group 7 to generate significant amounts of hydrogen peroxide may be important in pathogenesis. PMID- 12576594 TI - Transcriptional analysis of the 5' terminus of the flp fimbrial gene cluster from Actinobacillus actinomycetemcomitans. AB - Fresh isolates of the oral bacterial pathogen Actinobacillus actinomycetemcomitans exhibit a fimbriated, rough colony phenotype. Evidence suggests that the fimbrial subunit gene flp is part of a cluster of 14 genes (flp to tadG) thought to encode proteins involved in the synthesis, assembly and export of these fimbriae. To determine the transcriptional organization of the 5' terminus of this gene cluster, total RNA from rough and smooth phenotype variants of A. actinomycetemcomitans strain 283 were analysed by RT-PCR. Primers designed to amplify regions spanning gene junctions or multiple genes yielded amplicons at each individual gene junction from flp to tadD for both the rough and smooth variants. Semi-quantitative RT-PCR of the rcpA to tadZ amplicon revealed that significantly more mRNA was transcribed from the rough than the smooth variant. Longer amplicons encompassing flp to tadZ (3.9 kb) and tadA to tadD (2.1 kb) were also detected, but only from the rough variant. Rapid amplification of cDNA ends (RACE) was used to identify the 5' end of the mRNA containing flp. Antisense primers located within rcpC, orfB and flp-2 enabled amplification of a RACE product that was subsequently isolated and subcloned into pGEM-T. DNA sequencing indicated that the 5' end of the mRNA was located at a G or T nucleotide -102 to 101 nt upstream of flp. Corresponding sigma(70) consensus sequences were located at -10 (TATAAT) and -35 (TTGCAT) relative to the transcription start site. These data confirm that the flp gene cluster is an operon transcribed as a polycistronic message commencing from a G or T nucleotide located in the intergenic region upstream of flp. Promoter function of the flp upstream region was confirmed using a lacZ reporter gene construct transformed into Escherichia coli. RT-PCR analysis further suggests that although transcription does occur in both the rough and smooth variants, full-length transcripts are rapidly degraded or are significantly downregulated in the smooth variant. PMID- 12576595 TI - Ecosystem-dependent adaptive radiations of picocyanobacteria inferred from 16S rRNA and ITS-1 sequence analysis. AB - Small, coccoid and rod-shaped Synechococcus-type cyanobacteria with either phycoerythrin or phycocyanin as major accessory pigments were isolated from several large, temperate-zone lakes and the brackish Baltic Sea. The picocyanobacteria had two ribosomal operons with a long internal transcribed spacer (ITS-1) separating the 16S rDNA and 23S rDNA. A 16S rRNA-based phylogenetic analysis assigned all isolates to the picophytoplankton clade [sensu Urbach, E., Scanlan, D. J., Distel, D. L., Waterbury, J. B. & Chisholm, S. W. (1998). J Mol Evol 46, 188-201], which also comprises marine Synechococcus spp. and Prochlorococcus spp. The strains assorted to five paraphyletic clusters each containing two or more strains with 99.4-100 % 16S rRNA sequence identity. Five corresponding strain clusters were deduced from analysis of ITS-1 sequences. Sequence divergence in ITS-1 varied between 23 % in the most divergent and 1 % in the phylogenetically most conserved cluster. Clustered strains with low sequence divergence in ITS-1 were frequently isolated from a single ecosystem or hydrographically comparable lakes in the same region. They represent physiologically distinct ecotypes of species which, among other phenotypic variations, frequently differed in their major accessory pigments, the phycobiliproteins. The reproduction of the various pigment traits in different lineages was not correlated with the phylogenetic divergence deduced from 16S rRNA or ITS-1 sequences but rather seemed to be related to characteristics of the ecosystem and habitat from which the strains were isolated. The occurrence of a comparable spectrum of phenotypes in different lineages and ecosystems indicates that different strain clusters developed similar ecotypes during independent adaptive radiations. PMID- 12576596 TI - Characterization of a nitrate-respiring bacterial community using the nitrate reductase gene (narG) as a functional marker. AB - Bacterial cultures capable of reducing nitrate to nitrite, or of complete denitrification, were established from 5, 10, 15 and 20 cm depths of a freshwater sediment. Taxonomic analysis of the 56 isolates using 16S rRNA gene sequences revealed an unexpected species richness, which included representatives of the gamma-Proteobacteria, Bacillus spp., Staphylococcus spp. and members of the Actinobacteria. Gram-positive species tended to predominate in the lower depths of the sediment, where there was evidence of active sulphate respiration. Sequences (from the narG gene) potentially encoding the catalytic subunit of the membrane-associated nitrate reductase were successfully amplified from 46 of the isolates, using a nested PCR with four degenerate primers. NarG sequences clustered into three major groupings that were supported by alternative phylogenetic analyses. The NarG sequences from Gram-positive isolates (according to rRNA gene phylogeny) clustered together within sequences from the low-G+C Gram positive bacteria. However, this cluster also included two sequences from members of the genus Pseudomonas. Another group contained mostly NarG sequences from the Proteobacteria (according to rRNA gene phylogeny), but also included five sequences from Gram-positive species. The third group of NarG sequences contained three sequences from Gram-positive species. Thus, the NarG-derived phylogeny is not entirely consistent with 16S rRNA-based taxonomy, precluding the use of the narG gene as a taxonomically useful tool for the characterization of nitrate respiring bacteria. Total DNA was also extracted from the four depth intervals of the sediment sample and used in similar narG amplifications. Most sequences amplified directly from environmental DNA clustered in the Gram-negative group, and none was in the predominantly Gram-positive group. The study also revealed a degree of spatial organization of a nitrate-respiring community in terms of both microbiology and narG sequences. PMID- 12576597 TI - Expressed sequence tag (EST) analysis of two subspecies of Metarhizium anisopliae reveals a plethora of secreted proteins with potential activity in insect hosts. AB - Expressed sequence tag (EST) libraries for Metarhizium anisopliae, the causative agent of green muscardine disease, were developed from the broad host-range pathogen Metarhizium anisopliae sf. anisopliae and the specific grasshopper pathogen, M. anisopliae sf. acridum. Approximately 1,700 5' end sequences from each subspecies were generated from cDNA libraries representing fungi grown under conditions that maximize secretion of cuticle-degrading enzymes. Both subspecies had ESTs for virtually all pathogenicity-related genes cloned to date from M. anisopliae, but many novel genes encoding potential virulence factors were also tagged. Enzymes with potential targets in the insect host included proteases, chitinases, phospholipases, lipases, esterases, phosphatases and enzymes producing toxic secondary metabolites. A diverse array of proteases composed 36 % of all M. anisopliae sf. anisopliae ESTs. Eighty percent of the ESTs that could be clustered into functional groups had significant matches (E<10(-5)) in other ascomycete fungi. These included genes reported to have specific roles in pathogens with plant or vertebrate hosts. Many of the remaining ESTs had their best BLAST match among animal, plant and bacterial sequences. These include genes with plant and microbial counterparts that produce potent antimicrobials. The abundance of transcripts discovered for different functional groups varied between the two subspecies of M. anisopliae in a manner consistent with ecological adaptations of the two pathogens. By hastening gene discovery this project has enhanced development of improved mycoinsecticides. In addition, the M. anisopliae ESTs represent a significant contribution to the extensive database of sequences from ascomycetes that are saprophytes or plant and vertebrate pathogens. Comparative analyses of these sequences is providing important information about the biology and evolutionary history of this clade. PMID- 12576598 TI - RAM2: an essential gene in the prenylation pathway of Candida albicans. AB - Prenylation is a post-translational protein modification process that results in correct protein localization to membranes in the cell. It is mediated by at least three prenyltransferases (PTFs) in eukaryotic cells. The RAM2 gene encodes the common alpha-subunit of two PTFs, farnesyltransferase (FTase) and geranylgeranyltransferase I (GGTase I). In this study, the RAM2 gene of the prenylation pathway in the opportunistic pathogen Candida albicans was analysed. The heterozygote trisomy test was used to demonstrate that RAM2 is essential to the viability of C. albicans. RAM2 mRNA in the yeast and hyphae growth states was not detected by Northern blot analysis, but was detected by RT-PCR. Drugs that inhibit mammalian PTFs do not alter Candida cell growth, but they do inhibit FTase and GGTase I activities in cell-free enzyme assays. The data from genetic studies and cell-free enzyme assays suggest that the drugs may not have access to the Candida cytoplasm. The regulation of PTF subunits was also examined in a strain in which RAM2 was under the control of a regulable promoter. Overall, this study demonstrated that RAM2 is essential to C. albicans, indicating that protein prenylation is an indispensable cellular process in this yeast. PMID- 12576599 TI - Differential expression of Candida albicans phospholipase B (PLB1) under various environmental and physiological conditions. AB - Candida is the fourth most common organism responsible for bloodstream infections in many intensive care units, with Candida albicans being the most predominant species isolated in such cases. It has previously been shown that candidal phospholipase B, encoded by the PLB1 gene, is an important virulence factor for C. albicans pathogenesis. In this study, the effects of environmental factors (carbohydrate source and pH) and physiological conditions (serum, phospholipids and temperature) on the expression of PLB1 by C. albicans cells grown in rich [Sabouraud dextrose broth (SB) or yeast extract/peptone/dextrose] or chemically defined [Lee's, RPMI-1640 or yeast nitrogen base (YNB)] media were investigated. Northern blot analyses revealed that PLB1 mRNA was expressed in C. albicans cells grown in rich media at 30 degrees C but not at 37 degrees C. However, the protein Plb1p was detected in fungal cells growing at 37 degrees C in SB, as determined by Western blot analysis, indicating that although the mRNA for this gene was not detected, the actual gene product was present at this temperature. Expression of PLB1 was detected in cells grown in YNB/glucose at 30 degrees C but not at 37 degrees C. However, growth of C. albicans in YNB/glucose supplemented with serum and phospholipids resulted in expression of PLB1 at 37 degrees C also. Additionally, acidic pH induced higher levels of PLB1 mRNA expression compared to neutral pH, while the morphological form of C. albicans did not have any influence on the expression of this gene. The studies described here show that the expression of PLB1 is regulated by nutritional supplementation, environmental factors and the growth phase of the C. albicans cells, as well as by physiological conditions. The differential expression of PLB1 in response to environmental factors may be correlated to host-specific components available to C. albicans during infection. PMID- 12576600 TI - Comparison of real-time PCR with SYBR Green I or 5'-nuclease assays and dot-blot hybridization with rDNA-targeted oligonucleotide probes in quantification of selected faecal bacteria. AB - PCR primers and hybridization probes were designed for the 16S rRNA genes of six bacterial species or groups typically present in human faeces or used in the dairy industry. The primers and probes were applied for quantification of the target bacterial genomes added in artificial DNA mixtures or faecal DNA preparations, using dot-blot hybridization and real-time PCR with SYBR Green I and TaqMan chemistries. Dot-blot hybridization with (33)P-labelled oligonucleotide probes was shown to detect a 10 % target DNA fraction present in mixed DNA samples. Applicability of the rDNA-targeted oligonucleotide probes without pre-enrichment of the 16S gene pool by PCR was thus limited to the detection of the predominant microbial groups. Real-time PCR was performed using a 96-well format and was therefore feasible for straightforward analysis of large sample amounts. Both chemistries tested could detect and quantify a subpopulation of 0.01 % from the estimated number of total bacterial genomes present in a population sample. The linear range of amplification varied between three and five orders of magnitude for the specific target genome while the efficiency of amplification for the individual PCR assays was between 88.3 and 104 %. Use of a thermally activated polymerase was required with the SYBR Green I chemistry to obtain a similar sensitivity level to the TaqMan chemistry. In comparison to dot blot hybridization, real-time PCR was easier and faster to perform and also proved to have a superior sensitivity. The results suggest that real-time PCR has a great potential for analysis of the faecal microflora. PMID- 12576603 TI - Chlamydia screening in the United Kingdom. PMID- 12576604 TI - Spending money to save money. PMID- 12576605 TI - Is use of antiretroviral therapy among homosexual men associated with increased risk of transmission of HIV infection? AB - BACKGROUND/OBJECTIVE: There is concern that use of highly active antiretroviral therapy (HAART) may be linked to increased sexual risk behaviour among homosexual men. We investigated sexual risk behaviour in HIV positive homosexual men and the relation between use of HAART and risk of HIV transmission. METHODS: A cross sectional study of 420 HIV positive homosexual men attending a London outpatient clinic. Individual data were collected from computer assisted self interview, STI screening, and clinical and laboratory databases. RESULTS: Among all men, sexual behaviour associated with a high risk of HIV transmission was commonly reported. The most frequently reported type of partnership was casual partners only, and 22% reported unprotected anal intercourse with one or more new partners in the past month. Analysis of crude data showed that men on HAART had fewer sexual partners (median 9 versus 20, p=0.28), less unprotected anal intercourse (for example, 36% versus 27% had insertive unprotected anal intercourse with a new partner in the past year, p=0.03) and fewer acute sexually transmitted infections (33% versus 19%, p=0.004 in the past 12 months) than men not on HAART. Self assessed health status was similar between the two groups: 72% on HAART and 75% not on HAART rated their health as very or fairly good, (p=0.55). In multivariate analysis, differences in sexual risk behaviour between men on HAART and men not on HAART were attenuated by adjustment for age, time since HIV infection. CD4 count and self assessed health status. CONCLUSION: HIV positive homosexual men attending a London outpatient clinic commonly reported sexual behaviour with a high risk of HIV transmission. However, behavioural and clinical risk factors for HIV transmission were consistently lower in men on HAART than men not on HAART. Although use of HAART by homosexual men with generally good health is not associated with higher risk behaviours, effective risk reduction interventions targeting known HIV positive homosexual men are still urgently needed. PMID- 12576606 TI - Maintaining patient access to GUM clinics: is it compatible with appointments? AB - OBJECTIVES: To determine whether a planned change from a walk-in service to a system in which 35% of appointments were prebooked and 65% obtained on the day, preserved access to the service for patients with, or at risk of, STIs. To describe patients' attitudes and behaviours in relation to accessing the clinic, and relate these to disease status and other epidemiological factors. To evaluate the effect of the change on clinical outcomes. To develop a tool for evaluating access to services. METHOD: A natural experiment was studied, in which a cohort of patients attending just before the change in appointments policy (phase 1) was compared with a cohort following the change (phase 2). A questionnaire was administered to all new patients, and linked to disaggregated epidemiological and demographic data and case notes. RESULTS: The age, ethnic, symptom status, and disease mix of the clinic did not change significantly, and more patients were seen in phase 2. Time from telephoning to being seen did not change. Under 25s and Afro-Caribbeans used prebooked appointments less than others. Partner notification improved and HIV testing rates increased, while staff preferred the new system. CONCLUSION: This specific appointment system in a GUM clinic preserved access for high risk groups. However, this may not generalise to systems with a higher proportion of prebooked appointments. PMID- 12576607 TI - Opportunistic screening for genital chlamydial infection. I: acceptability of urine testing in primary and secondary healthcare settings. AB - OBJECTIVES: To determine the acceptability of opportunistic screening for Chlamydia trachomatis in young people in a range of healthcare settings. DESIGN: An opportunistic screening programme (1 September 1999 to 31 August 2000) using urine samples tested by ligase chain reaction (LCR). Data on uptake and testing were collected and in-depth interviews were used for programme evaluation. SETTING: General practice, family planning, genitourinary medicine clinics, adolescent sexual health clinics, termination of pregnancy clinics, and women's services in hospitals (antenatal, colposcopy, gynaecology and infertility clinics) in two health authorities (Wirral and Portsmouth and South East Hampshire). Main participants: Sexually active women aged between 16 and 24 years attending healthcare settings for any reason. MAIN OUTCOME MEASURES: Uptake data: proportion of women accepting a test by area, healthcare setting, and age; overall population coverage achieved in 1 year. Evaluation data: participants' attitudes and views towards opportunistic screening and urine testing. RESULTS: Acceptance of testing by women (16-24 years) was 76% in Portsmouth and 84% in Wirral. Acceptance was lower in younger women (Portsmouth only) and varied by healthcare setting within each site. 50% of the target female population were screened in Portsmouth and 39% in Wirral. Both the opportunistic offer of screening and the method of screening were universally acceptable. Major factors influencing a decision to accept screening were the non-invasive nature of testing and treatment, desire to protect future fertility, and the experimental nature of the screening programme. CONCLUSIONS: An opportunistic model of urine screening for chlamydial infection is a practical, universally acceptable method of screening. PMID- 12576608 TI - Opportunistic screening for genital chlamydial infection. II: prevalence among healthcare attenders, outcome, and evaluation of positive cases. AB - OBJECTIVES: To determine the prevalence and treatment outcomes among young women screened opportunistically for genital Chlamydia trachomatis and to evaluate the impact of screening in those participating. DESIGN: An opportunistic screening programme (1 September 1999 to 31 August 2000) using urine samples, tested by ligase chain reaction (LCR). In-depth interviews were used for programme evaluation. SETTING: Screening was offered in two health authorities at general practice, family planning, genitourinary medicine (GUM), adolescent sexual health, termination of pregnancy clinics and women's services in hospitals (antenatal, colposcopy, gynaecology and infertility clinics). Main participants: Sexually active women (16-24 years) attending for any reason. MAIN OUTCOME MEASURES: Screening data: prevalence of infection by age and healthcare setting; proportion of positive patients attending for treatment. Evaluation data: participants' attitudes and views towards screening and follow up. RESULTS: In total, 16 930 women (16-24 years) were screened. Prevalence was higher in younger women (16-20) than those aged 21-24 years and was highly variable at different healthcare settings (range 3.4%-17.6%). Prevalence was approximately 9% in general practice. The role of the project health advisers in managing results and coordinating treatment of positive individuals was essential; the vast majority of all positives were known to be treated. Women felt that screening was beneficial. Improving awareness and education about sexually transmitted infections is required to alleviate negative reactions associated with testing positive for infection. CONCLUSIONS: Prevalence of infection outside GUM clinics is substantial and opportunistic screening using urine samples is an acceptable method of reaching individuals with infection who do not normally present at specialist clinics. PMID- 12576609 TI - Repeat chlamydia screening by mail, San Francisco. AB - OBJECTIVES: To assess the feasibility and acceptability of home screening for repeat chlamydial infection using urine test kits sent through the mail. METHODS: A letter offering home rescreening was mailed to 399 adults who previously tested positive for chlamydia. Kits were then mailed to anyone who did not actively decline. The home testing kits contained instructions on how to collect a urine specimen and return the specimen by mail. Specimens were tested with strand displacement amplification. A short survey asked individuals their level of concern about confidentiality, safety, and privacy of mail screening. RESULTS: Among the 313 potential test kit recipients, 22.4% responded. Response rates were highest among homosexual and bisexual men (38.6%), people 35 years or older (34.3%), and white people (34.6%). The overall positivity rate was 3.2% (2/63). In women 18-25 years old, the positivity was 13.3% (2/15). CONCLUSIONS: Home testing with mailed urine collection kits is feasible and an acceptable method to screen for recurrent chlamydial infection. Young women would probably benefit most because of their higher rates of reinfection and risk for sequelae. PMID- 12576611 TI - Adolescence and other risk factors for Chlamydia trachomatis genitourinary infection in women in Melbourne, Australia. AB - OBJECTIVE: To establish the prevalence of and risk factors for Chlamydia trachomatis infection to determine the role of universal versus targeted testing. METHODS: A prospective study of 1107 women attending two sexual and reproductive health clinics in Melbourne, Australia, was carried out. A questionnaire was used to establish risk factors. Urine samples were tested for C. trachomatis by PCR. The main outcome measures were prevalence of and risk factors for C. trachomatis infection. RESULTS: Of 1107 recruitable women, 851 (76.9%) consented and were successfully tested. C. trachomatis was detected in 18 (4.8% (95% CI 2.9 to 7.5)) of 373 women in the inner city and eight (1.7% (95% CI (0.7 to 3.3)) of 478 women in the suburban clinic. Of women under 25 years, 17 (6.2% (95% CI 3.7 to 9.8)) of 273 in the inner city in contrast with three (1.7% (95% CI 0.4 to 5.0)) of 174 in the suburban clinic were infected. In the inner city clinic, age under 25 years (OR 5.4 (95% CI 0.7 to 41.5)), vaginal discharge (OR 4.1 (95% CI 1.5 to 11.1)), and recent change of sexual partner (OR 4.6 (95% CI 1.6 to 12.9)) were associated with C. trachomatis. In contrast, in the suburban clinic, only vaginal discharge (OR 3.5 (95% CI 0.9 to 14.3)) and recent change of sexual partner (OR 3.4 (95% CI 0.8 to 15.7)) were identified as risk factors. Multivariate analysis showed that recent change of partner (OR 4.5 (95% CI 1.5 to 13.8)) was the most strongly associated independent risk factor for infection in the inner city clinic. CONCLUSION: The high prevalence of C. trachomatis indicates that universal testing should be undertaken in the inner city clinic. Young age may not be a risk factor for C. trachomatis in more affluent populations with lower prevalence rates. No risk factors were identified with sufficient sensitivity and specificity to be useful for targeted testing. Prevalence and identifiable risk factors for C. trachomatis are not transferable between populations, even in the same city. PMID- 12576613 TI - Determinants of disclosure of genital herpes to partners. AB - OBJECTIVE: To identify factors which determine whether and when patients will disclose infection with genital herpes to sexual partners. METHODS: The sample was 26 women and 24 men attending a herpes clinic in a sexually transmitted disease clinic. Semistructured interviews yielded quantitative data and also qualitative data which were subjected to content analysis. RESULTS: Characteristics of partners were very important in determining whether disclosure occurred. Respondents were less likely to tell partners regarded as casual. Perception of the likely reaction of partners was important in deciding whether to tell. Many respondents assumed that they were not infectious if they were not currently having an attack or if they were taking antiviral medication. The decision whether to tell tended to be based on considerations of likely discovery and of honesty towards the partner rather than control of transmission. Of patient characteristics only self rated depressed mood was related to disclosure to the most recent partner. CONCLUSIONS: Perception of the partner and anticipated partner response is crucially important in determining whether and when disclosure of genital herpes infection occurs. PMID- 12576612 TI - Genital herpes serotesting: a study of the epidemiology and patients' knowledge and attitude among STD clinic attenders in Coventry, UK. AB - OBJECTIVES: To examine the seroprevalence and correlates of antibodies to herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2), and to assess patients' knowledge and attitude towards genital herpes infection and its serotesting, before and after counselling. METHODS: A cross sectional study among genitourinary medicine (GUM) clinic attenders in Coventry, a UK metropolitan city. Participants were asked to complete a self administered questionnaire before and after counselling. Patients were counselled before testing and after receiving the result. A commercially available enzyme immunoassay (EIA) was used to identify HSV-1 and HSV-2 antibodies (Gull/Meridian EIA). RESULTS: 223 patients participated in the study (97% of eligible patients). Overall, prevalence of HSV 2 antibody was 43/216 (20%) (19/103, 18% for males and 24/113 (21%) for females, p=0.61) while prevalence of HSV-1 antibody was 129/215 (60%) (60% for both sexes, p=0.91). In the multivariate analysis HSV-2 seropositivity was higher among black people and those with a history of genital herpes. HSV-1 seropositivity was independently associated with less education, increased years of sexual activity (between 14-25 years), and history of cold sores. The majority of patients wanted this serotesting to be available in the clinic (204/222 (92%) before and 216/218 (99%) after counselling, p=0.0003) and 97% accepted the test when offered. Only three patients regretted having the test and four patients contacted the department within 6 months of receiving the results for more counselling. CONCLUSION: The vast majority of the study population not only wanted to be tested, but accepted the test when offered. HSV-2 infection is common and largely unrecognised among our study population. The psychological impact of introducing type specific HSV serological testing in a clinical setting seems to be minimal. Counselling could improve patients' awareness of the infection and the acceptability of the test and its results. PMID- 12576615 TI - Advances in the prevention and treatment of paediatric HIV infection in the United Kingdom. PMID- 12576614 TI - Estimating the costs and benefits of screening monogamous, heterosexual couples for unrecognised infection with herpes simplex virus type 2. AB - OBJECTIVES: Herpes simplex virus type 2 (HSV-2) is the most common cause of ulcerative genital disease in the United States, but infection is commonly unrecognised. Serological screening tests could identify discordantly infected couples and permit targeted interventions to limit HSV-2 transmission. Our objective was to evaluate the projected cost effectiveness of strategies to prevent HSV-2 transmission in couples with no history of HSV-2 infection. METHODS: We created a mathematical model to simulate the natural history and costs of HSV-2 transmission, and the expected impact of HSV-2 prevention strategies in monogamous, heterosexual couples. Strategies evaluated included (i) no screening; (ii) universal condom use; and (iii) serological screening for HSV 2 with condom use targeted to discordant couples. Screening tests considered included western blot (WB), ELISA, and ELISA with confirmation of positive test results using WB (ELISA-->WB). RESULTS: Compared to no screening, the use of ELISA-->WB prevented 38 future infections per 1000 couples, with a cost effectiveness ratio of $8200 per infection averted. The use of WB in all couples had an incremental cost effectiveness ratio of $63 600 per infection averted. Strategies of ELISA alone and universal condom use were not cost effective. The cost effectiveness of ELISA-->WB improved with increasing prevalence of HSV-2, but worsened with decreasing condom compliance. Screening with ELISA alone was a reasonable strategy only when ELISA specificity increased to 99%. CONCLUSIONS: Serological screening for unrecognised HSV-2 infection in monogamous, heterosexual couples is expected to decrease the incidence of HSV-2 infection, but increase healthcare costs. For couples choosing to be screened, a two step testing strategy (ELISA-->WB) is recommended. Recommendations for a national policy to conduct serological screening will depend on the value placed on averting an incident HSV-2 infection. PMID- 12576616 TI - Detection of mixed infection of Neisseria gonorrhoeae. AB - We report the first published evidence, to our knowledge, of the demonstration of the potential existence of mixed infections of Neisseria gonorrhoeae in vivo through the use of a highly discriminatory genotypic technique. PMID- 12576617 TI - HIV treatment adherence and unprotected sex practices in people receiving antiretroviral therapy. AB - BACKGROUND: Poor HIV treatment adherence can result in the development of drug resistant strains of HIV and HIV positive people may transmit drug resistant virus to their sex partners. OBJECTIVE: To examine the association between HIV treatment adherence and sexual risk behaviour practices in people living with HIV AIDS. METHODS: Surveys and interviews with 255 men and women living with HIV and receiving antiretroviral therapy. RESULTS: People who were currently taking antiretroviral medications and missed at least one dose of their medications in the past week scored significantly higher on a hopelessness scale and reported more current use of marijuana. People who had been non-adherent also reported significantly more sex partners, greater rates of unprotected vaginal intercourse, and less protected sex behaviours including less protected sex with partners who were HIV negative or of unknown HIV status. CONCLUSIONS: Associations between HIV treatment adherence and sexual transmission risk behaviours indicate the need for comprehensive and integrated health behaviour interventions for people living with HIV-AIDS. PMID- 12576618 TI - Social capital, poverty, and income inequality as predictors of gonorrhoea, syphilis, chlamydia and AIDS case rates in the United States. AB - BACKGROUND: Social capital has been related to a number of important public health variables such as child welfare, mortality, and health status. However, the relation of social capital to infectious diseases has received relatively little attention. The relation of social capital to health measures is often posited to be related to the key societal variables of poverty and income inequality. Therefore, any exploration of the correlation between social capital and infectious diseases should also include examination of the association with poverty and income inequality. OBJECTIVE: This study examined the state level association between social capital, poverty, income inequality, and four infectious diseases that have important public health implications given their long term sequelae: gonorrhoea, syphilis, chlamydia, and AIDS. METHOD: A state level, correlational analysis (including bivariate linear correlational analysis, and multivariate linear stepwise regression analysis) was carried out. 1999 state level rates of gonorrhoea, syphilis, chlamydia, and AIDS were the main outcome measures. RESULTS: In bivariate analyses, poverty was significantly correlated with chlamydia; income inequality was significantly correlated with chlamydia and AIDS case rates; and social capital was significantly correlated with all outcome measures. In stepwise multiple regression analyses, social capital was always the strongest predictor variable. CONCLUSIONS: These results suggest that social capital is highly predictive of at least some infectious diseases. The results indicate the need for further research into this relation, and suggest the potential need for structural interventions designed to increase social capital in communities. PMID- 12576619 TI - Circumcision practice patterns in South Korea: community based survey. AB - OBJECTIVES: To assess the prevalence of circumcision in South Korean young men dwelling in the community, investigate attitudes and perspectives about circumcision, and how they perceive physician involvement in the decision process. METHODS: Between May and November 2001, this cross sectional survey was performed. Of 27 202 men aged 20 years dwelling in the community of Choong-chung South Province, 2700 were randomly selected at a 10.0% sampling fraction after a sampling process by census district and a total of 1742 (64.5%) agreed to participate in the study. These subjects completed self administered questionnaires and we included 1674 men (a response rate 62.0%) in the study. RESULTS: The overall proportion of circumcised was 1306 (78.0%) and an additional 192 (11.5%) wished to be circumcised later. Circumcision was carried out mostly during their elementary and middle school years. Of men circumcised, the decision whether to circumcise was most often made by their parents. Of the subjects, 75.0% believed that circumcision is necessary, while 2.9% believed it to be unnecessary. Among those who believed circumcision to be necessary, the most common reason was to improve penile hygiene (89.1%). CONCLUSIONS: Our results indicate a positive attitude toward circumcision in South Korean men, linking it with hygienic practices. Circumcision in South Korea depends on the perpetuation of cultural beliefs that support it. PMID- 12576620 TI - Chancroid: clinical manifestations, diagnosis, and management. AB - Chancroid is a sexually transmitted disease (STD) caused by the Gram negative bacterium Haemophilus ducreyi and is characterised by necrotising genital ulceration which may be accompanied by inguinal lymphadenitis or bubo formation. H ducreyi is a fastidious organism which is difficult to culture from genital ulcer material. DNA amplification techniques have shown improved diagnostic sensitivity but are only performed in a few laboratories. The management of chancroid in the tropics tends to be undertaken in the context of syndromic management of genital ulcer disease and treatment is usually with erythromycin. A number of single dose regimens are also available to treat H ducreyi infection. Genital ulceration as a syndrome has been associated with increased transmission of human immunodeficiency virus (HIV) infection in several cross sectional and longitudinal studies. Effective and early treatment of genital ulceration is therefore an important part of any strategy to control the spread of HIV infection in tropical countries. PMID- 12576623 TI - Chaperoning in genitourinary medicine clinics. PMID- 12576624 TI - Enhanced risk of HIV sexual transmission during structured treatment interruption. PMID- 12576626 TI - First, do not harm: also an issue in NAA assay diagnostics for chlamydial infection. PMID- 12576627 TI - Hepatitis C testing in HIV infected patients. PMID- 12576628 TI - Geographical focusing: an intervention to address increased risk for sexually transmitted diseases during repatriation and resettlement in post-war Mozambique. PMID- 12576630 TI - A novel research approach in sex on premises venues (SOPV): objective measure of sexual behaviour and low level intrusion to patrons. PMID- 12576631 TI - Prevalence of and risk factors for hepatitis C virus infection among STD clinic clientele in Miami, Florida. AB - OBJECTIVES: Hepatitis C virus (HCV) is the most common chronic blood borne viral infection in the United States. We assessed the HCV prevalence, risk factors, and sensitivity of the Centers for Disease Control and Prevention's (CDC) routine screening criteria among clients of a large urban sexually transmitted disease (STD) clinic. METHODS: Participants were recruited from a public STD clinic in Miami, Florida, and were interviewed regarding known and potential risk factors. The survey assessed CDC screening criteria, as well as other risk factors (for example, intranasal drug use, history of incarceration, exchanging sex for money, number of lifetime sex partners, and history of an STD). Testing was done by enzyme immunoassay (EIA) and confirmed by recombinant immunoblot assay (RIBA). RESULTS: The prevalence of anti-HCV positivity was 4.7%. Four variables were significantly associated with being anti-HCV positive, independent of confounding factors. These included injection drug use (odds ratio (OR) = 31.6; 95% confidence intervals (CI) 11.0 to 90.5); history of incarceration (OR = 3.0; 95% CI 1.1 to 8.1); sexual contact with an HCV positive person (OR 12.7; 95% CI 2.5 to 64.7); and older age (OR 1.4; 95% CI 1.2, 1.6). The sensitivity of CDC's routine screening criteria was 69% and specificity was 91%. CONCLUSIONS: The prevalence of anti-HCV in this clinic was similar to that determined in studies of comparable populations. Having sexual contact with an HCV positive person and history of incarceration were independently associated with being anti-HCV positive. CDC's screening criteria identified approximately two thirds of the anti-HCV positive participants. PMID- 12576632 TI - Toward and beyond lysosomes. PMID- 12576633 TI - Hrs and endocytic sorting of ubiquitinated membrane proteins. AB - Endocytosed receptors are either recycled to the plasma membrane or trapped within intralumenal vesicles of multi-vesicular bodies for subsequent degradation in lysosomes. How the cell is able to sort receptors in endosomes has so far been largely unknown. The hepatocyte growth factor regulated tyrosine kinase substrate, Hrs, is an essential protein that has been implicated in cell signalling and intracellular membrane trafficking. Very recently, several reports have demonstrated a role for Hrs in endocytic sorting of ubiquitinated membrane proteins. Here, we review current knowledge about how Hrs recognises ubiquitinated cargo that is destined for lysosomal degradation, and how Hrs may act as a key regulator of the molecular machinery involved in receptor sorting and multivesicular body formation. PMID- 12576634 TI - Autophagy in yeast: a review of the molecular machinery. AB - Autophagy is a membrane trafficking mechanism that delivers cytoplasmic cargo to the vacuole/lysosome for degradation and recycling. In addition to non-specific bulk cytosol, selective cargoes, such as peroxisomes, are sorted for autophagic transport under specific physiological conditions. In a nutrient-rich growth environment, many of the autophagic components are recruited for executing a biosynthetic trafficking process, the cytoplasm to vacuole targeting (Cvt) pathway, that transports the resident hydrolases aminopeptidase I and alpha mannosidase to the vacuole in Saccharomyces cerevisiae. Recent studies have identified pathway-specific components that are necessary to divert a protein kinase and a lipid kinase complex to regulate the conversion between the Cvt pathway and autophagy. Downstream of these proteins, the general machinery for transport vesicle formation involves two novel conjugation systems and a putative membrane protein complex. Completed vesicles are targeted to, and fuse with, the vacuole under the control of machinery shared with other vacuolar trafficking pathways. Inside the vacuole, a potential lipase and several proteases are responsible for the final steps of vesicle breakdown, precursor enzyme processing and substrate turnover. In this review, we discuss the most recent developments in yeast autophagy and point out the challenges we face in the future. PMID- 12576635 TI - Autophagosome formation in mammalian cells. AB - Macroautophagy is an intracellular degradation system for the majority of proteins and some organelles. The molecular mechanism of autophagy has been extensively studied using the yeast, Saccharomyces cerevisiae, during these past 10 years. These studies suggested that the molecular machinery of autophagosome formation is well conserved from yeast to higher eukaryotes. Identification and characterization of the mammalian counterparts of the yeast autophagy proteins has facilitated our understanding of mammalian autophagy, particularly of autophagosome formation. These findings are now being applied to studies on the physiological roles of autophagy in mammals. PMID- 12576636 TI - Diversity of signaling controls of macroautophagy in mammalian cells. AB - Macroautophagy is a major lysosomal catabolic process conserved from yeast to human. The formation of autophagic vacuoles is stimulated by a variety of intracellular and extracellular stress situations including amino acid starvation, aggregation of misfolded proteins, and accumulation of damaged organelles. Several signaling pathways control the formation of autophagic vacuoles. As some of them are engaged in the control of protein synthesis or cell survival this suggests that macroautophagy is intimately associated with the execution of cell proliferation and cell death programs. Whether or not these different signaling pathways converge to a unique point to trigger the formation of autophagic vacuole remains an open question. PMID- 12576637 TI - Lysosome-related organelles: a view from immunity and pigmentation. AB - Lysosomes are ubiquitous organelles that carry out essential household functions. Certain cell types, however, contain lysosome-related organelles with specialized functions. Their specialized functions are usually reflected by specific morphological and compositional features. A number of diseases that develop due to genetic mutations, pathogen exposure or cell transformation are characterized by dysfunctional lysosomes and/or lysosome-related organelles. In this review we highlight adaptations and malfunction of the endosomal/lysosomal system in normal and pathological situations with special focus on MHC class II compartments in antigen presenting cells and melanosomes in pigment cells. PMID- 12576638 TI - Molecular dynamics of Aurora-A kinase in living mitotic cells simultaneously visualized with histone H3 and nuclear membrane protein importinalpha. AB - Aurora-A is known to be a mitotic kinase required for spindle assembly. We constructed a human stable cell-line in which Aurora-A, histone H3 and importinalpha were differentially expressed as fusions to green, cyan, and red fluorescent proteins (GFP, CFP and DsRed). In interphase cells, GFP-Aurora-A was localized in the centrosome. Its molecular behavior in living mitotic cells was extensively analyzed by an advanced timelapse image analyzing system. In G2 phase, duplicated centrosomal dots of Aurora-A separated and moved to the opposite poles, a process requiring 18 min. In prophase, the Aurora-A dots approached closer and the nuclear membrane of DsRed-importinalpha beneath them became thick and invaginated, resulting in a "dumb-bell" shaped nucleus with condensed chromatin. As the importinalpha membrane further shrank and disappeared, the condensed chromatin was excluded from the nucleus and the Aurora A dots grew rapidly into a spindle-like structure. Congression of mitotic chromosomes continued for 20-50 min until they were properly aligned at the spindle equator and then the sister chromatids started to segregate, taking 4-6 min for them to reach the poles. An importinalpha membrane reappeared around the surface of chromatin 10 min after anaphase onset. Aurora-A gradually decreased in size in telophase and returned to the surface of the newly formed small sister nuclei. These observations showed that the morphological change of Aurora-A was cooperated with the breakdown and reformation of nuclear membrane. Immunostaining with anti-alpha or gamma-tubulin further indicated that Aurora-A was involved in the formation of mitotic spindle in metaphase as well as the subsequent chromosome movement in anaphase. PMID- 12576639 TI - Endocytic pathway in mouse cardiac cells. AB - Primary cultures of heart muscle cells provide powerful tools for cardiac cell biological research that permits both physiological and biochemical approaches. In the present study we analyzed the endocytosis of cardiac cells and presented morphological characterization of the endocytic machinery using markers, which enabled us to follow the fluid-phase, receptor-mediated endocytosis and the internalization of large particles. Our results demonstrated the route of the internalized cargo to early endosomes followed or not by its discharge in the late compartments. We also confirmed the ability of cardiac muscle cells to ingest large particles such as the mannosylated ligand zymosan A, and even internalize whole eukaryotic cells such as the protozoan parasite Trypanosoma cruzi. Since endocytosis is involved in many important cellular functions, the present work contributes to the knowledge of possible additional roles played by cardiac muscle cells besides their well known ability to act as physically energetic cells in the body, constantly contracting without tiring. PMID- 12576640 TI - Spatial localization of mono-and diphosphorylated myosin II regulatory light chain at the leading edge of motile HeLa cells. AB - Nonmuscle myosin II activity is regulated by phosphorylation of the myosin II regulatory light chain (MRLC) at Ser19 or at both Thr18 and Ser19, and the phosphorylation of MRLC promotes the contractility and stability of actomyosin. To analyze the states of MRLC phosphorylation at the leading edge in the motile HeLa cells, we have examined the subcellular distribution of monophosphorylated or diphosphorylated form of MRLC using a confocal microscope. The cross-sectional imaging revealed that monophosphorylated MRLC distributed throughout the cortical region and the leading edge, but its fluorescent signal was much stronger at the leading edge. This distribution pattern of monophosphorylated MRLC was almost identical to those of myosin II and F-actin. On the other hand, diphosphorylated MRLC is localized at the base of leading edge, spatially very close to the substrate, and colocalized with F-actin in part at the base of filopodia. Diphosphorylated MRLC was hardly detectable at the tip of filopodia and the cell cortical region, where monophosphorylated MRLC was clearly detected. These localization patterns suggest that myosin II is activated at the leading edge, especially at the base but not the tip of filopodia in motile cells. Next, we analyzed the cells expressing GFP-tagged recombinant MRLCs. Expression of GFP tagged diphosphorylatable and monophosphorylatable MRLCs led to a significant increase in the filopodial number, compared with the cells expressing nonphosphorylatable MRLC. This result indicated that expression of phosphorylatable MRLC enhances the formation of filopodia at the wound edge. PMID- 12576641 TI - Anti-tack action of polyvinylpyrrolidone on hydroxypropylmethylcellulose solution. AB - The anti-tack action of polyvinylpyrrolidone (PVP) on hydroxypropylmethylcellulose (HPMC) solution was elucidated using a probe test method. The influence of PVP of varying molecular weights at various PVP concentrations and solution temperatures on the tackiness of HPMC solution was studied. The viscosity, surface tension, cloud point and solution spectroscopy of HPMC solutions and glass transition temperature of HPMC films, with and without PVP, were investigated. The tackiness of HPMC solutions in response to the addition of PVP, at different concentrations of HPMC and using HPMC with varying contents of hydroxypropyl/methoxyl substitution, was also evaluated. PVP is a commonly used binder and adhesive. However, it reduced the tack of the HPMC solution when used at low concentrations, without affecting the state of hydration of HPMC. Lower molecular weight PVP was more effective as an anti-tack agent owing to suitable hydrodynamic size to intersperse among the HPMC chains. The degree of reduction in tack values was more pronounced for HPMC that showed a greater extent of interaction between polymer chains such as when high concentration of HPMC or low solution temperature was employed. This indicated that the tack reduction property of PVP relied on its ability to interact with the HPMC chains. The profile of reduction in tack values was affected by the contents of HPMC substitution and was a result of net reduction in the extent of hydrogen bonding between HPMC chains. It was significantly correlated to the changes of viscosity and surface tension of the HPMC solutions but not to the glass transition temperatures of the polymers prepared as solid films. The results suggested that the anti-tack action of PVP was attributed to its ability to interact with HPMC chains in the aqueous medium and consequently to reduce the extent of HPMC-HPMC bonding. PMID- 12576642 TI - Development of 5-[(3-aminopropyl)phosphinooxy]-2-(hydroxymethyl)-4H-pyran-4-one as a novel whitening agent. AB - A stable derivative of kojic acid, 5-[(3-aminopropyl)phosphinooxy]-2 (hydroxymethyl)-4H-pyran-4-one (Kojyl-APPA), was synthesized in good yield. The effects of Kojyl-APPA on tyrosinase activity and melanin synthesis were investigated. Kojyl-APPA showed tyrosinase inhibition effect (30%) in situ, but not in vitro. Kojyl-APPA inhibited tyrosinase activity significantly at 24 h after treatment in normal human melanocytes. It means that Kojyl-APPA is not a direct inhibitor of tyrosinase itself, but it is converted to a potential inhibitor kojic acid enzymatically in cells. In addition, Kojyl-APPA decreased melanin content to 75% of control in melanoma cells and decreased neomelanin synthesis to 43% of control in normal human melanocytes. Its permeation through skin increased by about 8 times as compared with kojic acid. PMID- 12576643 TI - Synthesis of potent and selective inhibitors against the proliferation of human coronary artery smooth muscle cells. AB - A series of diarylamide urea derivatives were synthesized and evaluated for their inhibitory activities against human coronary artery smooth muscle cells (SMCs) and human coronary artery endothelial cells (ECs). Compound 2h was much superior to Tranilast, in terms of both the potency of its inhibitory activity toward the proliferation of SMCs and the cell selectivity. PMID- 12576644 TI - Synthesis of eosinophil infiltration inhibitors with antihistaminic activity. AB - A series of [1, 2, 4]triazolo[1, 5-b]pyridazines (5) and imidazo[1, 2 b]pyridazines (6) having cyclic amines was synthesized and evaluated for antihistaminic activity and inhibitory effect on eosinophil infiltration. When a piperidine or a piperazine containing a benzhydryl group and a suitable spacer was incorporated at the 6-position, the fused pyridazines were found to exhibit both antihistaminic activity and an inhibitory effect on eosinophil chemotaxis. Above all, 6a showed potent antihistaminic activity, but little blockade of central H(1) receptors in contrast with its complete blockade of peripheral H(1) receptors as determined by an ex vivo binding assay. Furthermore, 6a inhibited eosinophil infiltration of the skin caused by a topical antigen challenge in sensitized guinea pigs, while an antihistamine terfenadine was not effective. After the pharmacokinetic study, 6a was found to be rapidly hydrolyzed to 6o, which was also orally active. Compound 6o, 2-[6-[[3-[4 (diphenylmethoxy)piperidino]propyl]amino]imidazo[1, 2-b]pyridazin-2-yl]-2 methylpropionic acid dihydrate (TAK-427), having both antihistaminic and antiinflammatory activity, is currently undergoing clinical trials as a therapeutic agent for atopic dermatitis and allergic rhinitis. PMID- 12576645 TI - Pentacyclic triterpenoids from the aerial parts of Lantana camara. AB - Three new pentacyclic triterpenoids, camaryolic acid (1), methylcamaralate (2) and camangeloyl acid (3) and six known compounds beta-sitosterol 3-O-beta-D glucopyranoside (4), octadecanoic acid (5), docosanoic acid (6), palmitic acid (7), camaric acid (8) and lantanolic acid (9) were isolated from the aerial parts of Lantana camara. Structures of the new compounds were elucidated by spectroscopic and chemical methods. PMID- 12576646 TI - Studies on non-thiazolidinedione antidiabetic agents. 2. Novel oxyiminoalkanoic acid derivatives as potent glucose and lipid lowering agents. AB - We previously reported that (Z)-2-(4-[(5-methyl-2-phenyl-1, 3-oxazol-4 yl)methoxy]benzyloxyimino)-2-(4-phenoxyphenyl)acetic acid (3) showed potent glucose and lipid lowering effects in genetically obese and diabetic mice, KKA(y). This compound also showed transcriptional activity for peroxisome proliferator-activated receptor (PPAR)-gamma. We expanded on the structure activity relationships of oxyiminoalkanoic acid derivatives based on this transcriptional activity (in vitro). Insertion of a carbon chain between the imino carbon and the carboxyl moiety of (Z)-2-(4-[(5-methyl-2-phenyl-1, 3-oxazol 4-yl)methoxy]benzyloxyimino)-2-phenylacetic acid (2) resulted in a marked increase in transcriptional activity at PPARgamma. In vivo potencies of synthesized compounds, which showed strong functional activity at PPARgamma, were tested using KKA(y) mice. Among these compounds, (E)-4-(4-[(5-methyl-2-phenyl-1, 3-oxazol-4-yl)methoxy]benzyloxyimino)-4-phenylbutyric acid (27) exhibited marked glucose and lipid lowering activity while showing no significant body weight gain. Compound (27) (TAK-559) showed favorable pharmacokinetic properties with good absorption and duration, and was considered as an attractive candidate for further evaluation. PMID- 12576647 TI - Glycosides of Atractylodes japonica. AB - From the water-soluble portion of the methanol extract of the fresh rhizome of Atractylodes japonica, five new sesquiterpenoid glycosides, including a compound having a secoguaiane skeleton, and a new aromatic compound glycoside were isolated together with ten known compounds. Their structures were clarified by spectral investigation. PMID- 12576648 TI - Quantitative analysis of bilobalide and ginkgolides from Ginkgo biloba leaves and Ginkgo products using (1)H-NMR. AB - 1H-NMR spectrometry was applied to the quantitative analysis of the bilobalide, ginkgolides A, B, and C in Ginkgo biloba leaves and six kinds of commercial Ginkgo products without any chromatographic purification. The experiment was performed by the analysis of each singlet H-12, which were well separated in the range of delta 6.0-7.0 in the (1)H-NMR spectrum. However, the H-12 protons of bilobalide and ginkgolides may have overlapped with H-6 or H-8 protons of the Ginkgo flavonoids. Therefore, the optimum (1)H-NMR solvent for the analysis of the compound was selected through the evaluation of solvent effects on the resolution of these signals from the compounds. Acetone-d(6)-benzene-d(6) (50 : 50) was found to be the best one among the solvents evaluated. The quantity of the compounds was calculated by the relative ratio of the intensity of each compound to the known amount of internal standard (25 microgram), phloroglucinol. This method allows rapid and simple quantitation of underivatized bilobalide and ginkgolides in 5 min without any pre-purification steps. PMID- 12576649 TI - Pharmacological evaluation of some new 6-amino/methyl pyridine derivatives. AB - In the present study, a series of 2-substituted-pyridines were synthesized and characterized by IR, (1)H-NMR and Elemental Analysis. The compounds were assayed against seizures induced by maximal electro shock (MES) and pentylenetetrazole (scMet). Neurologic deficit was evaluated by the rotarod test. The decrease in the elevated motor activity by introceptive chemical stimuli (amphetamine antagonistic activity) was studied at the dose level of 25 and 50 mg/kg, antihistaminic and cardiac activity were also studied. All the compounds exhibited significant anticonvulsant activity. Compounds 2-(2-hydroxy-3 piperazinopropylamino)-6-aminopyridine, 2-[2-hydroxy-3-(1-imidazolyl)propylamino] 6-aminopyridine, 2-[2-(1-imidazolyl)ethylamino]-6-methylpyridine and 2-[2 (methylamino)ethylamino]-6-methylpyridine were most active of the series against MES-induced seizures. Compounds 2-[2-(phenylamino)ethylamino]-6-aminopyridine, 2 [2-[bis(2-hydroxyethyl)amino]ethylamino]-6-aminopyridine, 2-[2 (diethylamino)ethylamino]-6-methylpyridine and 2-[2-hydroxy-3-(1 imidazolyl)propylamino]-6-methylpyridine exhibited significant decrease in the elevated motor activity at the dose of 50 mg/kg. Remarkable sympathetic blocking activity was observed with 2-(2-hydroxy-3-piperazinopropylamino)-6-aminopyridine, 2-(2-hydroxy-3-morpholinopropylamino)-6-methylpyridine and 2-(2-hydroxy-3 piperazinopropylamino)-6-methylpyridine only. Compounds 2-[2 (diethylamino)ethylamino]-6-aminopyridine, 2-[2-[bis(2 hydroxyethyl)amino]ethylamino]-6-aminopyridine, and 2-[2 (diethylamino)ethylamino]-6-methylpyridine exhibited significant blocking of histamine induced contraction on guinea pig ileum. PMID- 12576650 TI - Nanoparticle formation of poorly water-soluble drugs from ternary ground mixtures with PVP and SDS. AB - Poorly water-soluble drugs N-5159, griseofulvin (GFV), glibenclamide (GBM) and nifedipine (NFP) were ground in a dry process with polyvinylpyrrolidone (PVP) and sodium dodecyl sulfate (SDS). Different crystallinity behavior of each drug during grinding was shown in the ternary Drug/PVP/SDS system. However, when each ternary Drug/PVP/SDS ground mixture was added to distilled water, crystalline nanoparticles which were 200 nm or less in size were formed and had excellent stability. Zeta potential measurement suggested that the nanoparticles had a structure where SDS was adsorbed onto the particles that were formed by the adsorption of PVP on the surface of drug crystals. Stable existence of crystalline nanoparticles was attributable to the inhibition of aggregation caused by the adsorption of PVP and SDS on the surface of drug crystals. Furthermore, the electrostatic repulsion due to the negative charge of SDS on a shell of nanoparticles could be assumed to contribute to the stable dispersion. PMID- 12576651 TI - Synthesis and characterization of 6-O-acyl-2-O-alpha-D-glucopyranosyl-L-ascorbic acids with a branched-acyl chain. AB - We previously reported the chemical synthesis of a series of novel monoacylated vitamin C derivatives, 6-O-acyl-2-O-alpha-D-glucopyranosyl-L-ascorbic acids (6 Acyl-AA-2G) possessing a straight-acyl chain of varying length from C(4) to C(18), as effective skin antioxidants. In this paper, we describe branched type of 6-Acyl-AA-2G derivatives (6-bAcyl-AA-2G) synthesized by use of a 2-branched chain fatty acid anhydride as an acyl donor. The stability of 6-bAcyl-AA-2G in neutral solution was much higher than that of 6-Acyl-AA-2G, while they were susceptible to enzymatic hydrolysis for exerting vitamin C effect. These branched derivatives as well as 6-Acyl-AA-2G increased the radical scavenging activity against 1, 1-diphenyl-2-picrylhydrazyl and the lipophilicity in octanol/water partitioning systems with increasing length of their acyl group. In addition, the 6-bAcyl-AA-2G derivative with an acyl chain of C(12), 6-bDode-AA-2G had the excellent solubility to various solvents, suggesting easy handling in cosmetic use. These characteristics of 6-bAcyl-AA-2G may be available for skin care application as an effective antioxidant. PMID- 12576652 TI - The first example of alpha-thiomagnesiums generated from dithioacetal monoxides with Grignard reagent; their properties and some synthetic applications. AB - Dithioacetal monoxides were synthesized from aldehydes and cyclohexanone, and reaction of the dithioacetal monoxides with Grignard reagents was investigated. The dithioacetal monoxide synthesized from alkylaldehyde and 4-chlorobenzenethiol reacted with i-PrMgCl to afford the desired alpha-thiomagnesium in high yield. The generated alpha-thiomagnesium was found to be stable at room temperature and to be useful in organic synthesis. In contrast to this, the dithioacetal monoxides derived from benzaldehyde and cyclohexanone did not give satisfactory results. PMID- 12576653 TI - Studies on the chemical constituents of stem bark of Millettia leucantha: isolation of new chalcones with cytotoxic, anti-herpes simplex virus and anti inflammatory activities. AB - Four new chalcone derivatives (1, 4, 7, 10) were isolated from the stem bark of Millettia leucantha KURZ (Leguminosae) along with two known ones (2, 6) and five known flavones (3, 5, 8, 9, 11). Structure elucidation and unambiguous assignment of the isolates were achieved with the aid of 1D and 2D NMR extensive studies. Correlation of 10 to 4 was successfully done by reduction with Et(3)SiH/CF(3)CO(2)H. Moderate cytotoxic activity was observed in chalcones (1, 10), whereas dihydrochalcones (4, 6) showed moderate anti-Herpes Simplex Virus (HSV) activity. Interestingly, flavone 8 showed significant anti-inflammatory effects inhibiting both cyclooxygenase (COX)-1 and -2. PMID- 12576654 TI - Two new flavonoids from Andrographis rothii. AB - Two new flavonoids, 5, 7, 2', 5'-tetramethoxyflavanone (1) and 5-hydroxy-7, 2' dimethoxyflavone (2), together with two known flavones, skullcapflavone I (3) and echioidin (4) were isolated from the whole plant of Andrographis rothii. The structures of the new compounds were established by extensive one- and two dimensional (1D- and 2D-) NMR spectral studies. PMID- 12576655 TI - A new coumestan from Tephrosia calophylla. AB - A new coumestan, tephcalostan (1) has been isolated from the whole plant of Tephrosia calophylla BEDD. together with two known flavonoids, 7-O methylglabranin (2) and kaempferol 3-O-beta-D-glucopyranoside (3). The structure of tephcalostan was elucidated as 5'-(R)-8, 9-methylenedioxy-5'-isopropenyl-4', 5'-dihydrofurano[2', 3':2, 3]coumestan by extensive one-and two-dimensional (1D- and 2D-)-NMR techniques including (1)H-(1)H correlation spectroscopy (COSY), heteronuclear single quantum coherence (HSQC), heteronuclear multiple bond connectivity (HMBC) and nuclear Overhauser enhancement spectroscopy (NOESY) experiments. PMID- 12576656 TI - A new ent-clerodane diterpene from the aerial parts of Baccharis gaudichaudiana. AB - A new ent-clerodane diterpene, named bacchariol (1) was isolated from the aerial parts of Baccharis gaudichaudiana DC. (Compositae), together with known ent clerodane diterpenes (2, 3), eight known flavonoids (4-11) and 3, 5 dicaffeoylquinic acid (12). Their structures were determined by spectroscopic analyses. Flavonoids (7, 8, 11) and 12 showed moderate scavenging activities toward 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals. PMID- 12576657 TI - A new triterpene glucosyl ester from the fruit of the blackberry (Rubus allegheniensis). AB - A new triterpene glucosyl ester, rubusside A, has been isolated from the fruit of the blackberry (Rubus allegheniensis PORT.) along with a known triterpene glucosyl ester, niga-ichigoside F1. The chemical structure of rubusside A was determined on the basis of spectroscopic data as well as chemical evidence. PMID- 12576658 TI - O-demethylation and sulfation of 7-methoxylated flavanones by Cunninghamella elegans. AB - Metabolism of 7-O-methylnaringenin (sakuranetin) by Cunninghamella elegans NRRL 1392 yielded naringenin and naringenin-4'-sulfate. C. elegans also converted 5, 3', 4'-trihydroxy-7-methoxyflavanone into eriodictyol-4'-sulfate. Furthermore, incubation of 5, 4'-dihydroxy-7, 3'-dimethoxyflavanone with the same fungus gave homoeriodictyol (5, 7, 4'-trihydroxy-3'-methoxyflavanone) and homoeriodicytol-7 sulfate. The structures of the new metabolites were established by spectral analysis including 2D-NMR, HR-ESI-FT-MS beside hydrolysis by acid. PMID- 12576659 TI - Diterpenoids from Amazonian crude drug of Fabaceae. AB - A new cassane diterpene was isolated from Amazonian crude drug, "Acapu, " the wood of Vouacapoua americana, together with two known cassane furanoditerpenes, (+)-vouacapenic acid and (+)-methyl vouacapenate. The structure was characterized as cassa-13(14), 15-dien-19-oic acid on the basis of spectroscopic evidence. PMID- 12576660 TI - Synthesis of granulatimide positional analogues. AB - The Stille coupling reaction of the stannylindole 13 with the 5-iodoimidazole derivative 14 (or 27) in the presence of PdCl(2)(PPh(3))(2) gave the corresponding indole-imidazole coupling product 15 (or 28), thereby affording a synthetic approach to 10-methylgranulatimide (7), 15-methylgranulatimide (11), and 10, 15-dimethylgranulatimide (12), as well as 10-methylisogranulatimide B (5). PMID- 12576661 TI - Bacopasides III-V: three new triterpenoid glycosides from Bacopa monniera. AB - Three new saponins, designated as bacopasides III, IV and V have been isolated from Bacopa monniera WETTST. and their structures have been elucidated as 3-O alpha-L-arabinofuranosyl (1-->2)-beta-D-glucopyranosyl jujubogenin (1), 3-O-beta D-glucopyranosyl (1-->3)-alpha-L-arabinopyranosyl jujubogenin (2) and 3-O-beta-D glucopyranosyl (1-->3)-alpha-L-arabinopyranosyl pseudojujubogenin (3) mainly on the basis of two dimensional (2D) NMR and other spectral analyses. PMID- 12576662 TI - Effects of natural polysaccharide addition on drug release from calcium-induced alginate gel beads. AB - Calcium-induced alginate gel beads (Alg-Ca) containing various polysaccharides, including an alginate hydrolysate, were prepared and the drug release profiles were investigated. Hydrocortisone (HC) was gradually released from Alg-Ca into the mimic gastric fluid, while in intestinal fluid, it was quickly released with the dissolution of Alg-Ca. However, with Alg-Ca containing 5% chitin (CT), dissolution of Alg-Ca was not observed, and release of HC showed apparent zero order kinetics. Furthermore, addition of the alginate hydrolysate altered the HC release profile for Alg-Ca. PMID- 12576663 TI - Asymmetric borane reduction of prochiral ketone using chiral bis(alpha, alpha diphenyl-2-pyrrolidinemethanol) carbonate. AB - Chiral bis(alpha, alpha-diphenyl-2-pyrrolidinemethanol) carbonate (DPP(2).H(2)CO(3)) is a useful asymmetric auxiliary for the asymmetric borane reduction of prochiral ketones. Chiral DPP(2).H(2)CO(3) is recoverable from the reaction and directly reusable for the reaction. The intermediate of KUR-1246, which we are developing as a new uterine relaxant, was synthesized using the methodology. PMID- 12576664 TI - Steric-control for the enantioselective hydrolysis of amino acid esters in hybrid membrane systems. AB - The enantioselective hydrolysis of the amino acid esters, p-nitrophenyl-N dodecanoyl-D(L)-phenylalaninates (C(12)-D(L)-Phe-PNP) catalyzed by active tripeptide, N-(benzyloxycarbonyl)-L-phenylalanyl-L-histidyl-L-leucine (Z PheHisLeu) in the presence of coaggregates (hybrid membranes) composed of native phospholipid, L-alpha-dimyristoylphosphatidylcholine (DMPC) and nonionic surfactant, polyoxyethylene (8) lauryl ether (C(12)(EO)(8)) was easily controlled by regulating the reaction temperature and changing the composition of coaggregates. Furthermore, excellent correlations were observed between the enantioselectivity in the hydrolysis of C(12)-D(L)-Phe-PNP catalyzed by Z PheHisLeu in the presence of coaggregates and physical properties of hybrid membranes. It is assumed that catalytic activities of tripeptide catalyst in hybrid membranes should be regulated by changing the microenvironments of reaction fields. PMID- 12576665 TI - Novel channel structure of bile acid-guest inclusion complex formed between ursodeoxycholic acid and phenanthrene. AB - The crystal structure of inclusion complex between ursodeoxycholic acid (UDCA), and phenanthrene has been determined. UDCA molecules formed hydrogen bond network to provide the channel structure along b axis, and phenanthrene molecules were accommodated in the cavity with a stoichiometry of 1 : 1 molar ratio. The channel structure observed in the UDCA-phenanthrene complex was significantly different from that of inclusion complex previously reported for deoxycholic acid (DCA) and cholic acid (CA). Because of the mesh-like hydrogen bond network, channel framework of UDCA could have less flexibility than that of DCA and CA. The difference of molecular state of phenanthrene was clearly observed in solid-state fluorescence measurement. PMID- 12576666 TI - Anti-diabetes effect of Zn(II)/carnitine complex by oral administration. AB - A novel bis(L-carnitinato)Zn(II) complex, Zn(car)(2)Cl(2), was prepared, and its insulinomimetic and antidiabetic activities were examined. The complex showed a tendency to lower the high blood glucose levels of KK-A(y) mice with type 2 diabetes mellitus when given by oral administration at a dose of 20 mg Zn/kg body weight for 16 d. In addition, the complex improved glucose tolerance ability when examined by the oral glucose tolerance test (1 g glucose/kg body weight). PMID- 12576667 TI - Gluco-indole alkaloids from Nauclea cadamba in Thailand and transformation of 3 alpha-dihydrocadambine into the indolopyridine alkaloid, 16 carbomethoxynaufoline. AB - Three monoterpenoid gluco-indole alkaloids, 3beta-isodihydrocadambine, cadambine, and 3alpha-dihydrocadambine, were isolated from Nauclea cadamba ROXB. growing in Thailand. The stereochemistry at C19 in 3beta-isodihydrocadambine was elucidated to be R by spectroscopic analysis. Treatment of 3alpha-dihydrocadambine with beta glucosidase in aqueous ammonium acetate solution gave an indolopyridine alkaloid, 16-carbomethoxynaufoline, and an unusually rearranged compound. PMID- 12576668 TI - Cytotoxic major saponin from tomato fruits. AB - A major novel steroidal alkaloid glycoside, possessing cytotoxic activity has been isolated from the fruits of Lycopersicon esculentum. PMID- 12576669 TI - Characterization of proteins responsive to gibberellin in the leaf-sheath of rice (Oryza sativa L.) seedling using proteome analysis. AB - In plants gibberellins (GAs) are responsible for triggering stem or internodal elongation. To comprehend the molecular basis of internodal elongation in rice, a proteomics approach using differentially displayed proteins on two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was carried out to identify the proteins expressed during the GA controlled leaf-sheath elongation response. Out of 352 protein spots detected on 2-D PAGE, 32 proteins showed modulation in the expression levels in GA3-treated leaf-sheath for 48 h as compared to control. These proteins were analyzed using protein sequencer and/or mass spectrometry in conjunction with the protein database to assign putative identities. The twin protein spots (LS079 and LS083), identified as calreticulin, showed different isoelectric points and expression level in GA3-treated leaf-sheath. The expression level of LS083 (pI 4.0) was down-regulated as compared to the up regulation of LS079 (pI 4.3). In the presence of GA3 and growth inhibitor, uniconazole and abscisic acid, respectively, no elongation in leaf-sheath occurred and calreticulin did not shift from LS083 to LS079. Over-expression of calreticulin in rice inhibited the callus regeneration and seedling growth. These results suggest that calreticulin is an important component in the GA signaling pathway that regulates rice seedling leaf-sheath elongation. PMID- 12576670 TI - Expression of adenovirus type 5 E1A in the methylotrophic yeast Pachia pastoris and the inhibitory effect on S-180 tumor growth. AB - The human adenovirus type 5 (Ad5) early-region 1A (E1A) proteins have been shown to have strong tumor-suppressive activities in human tumor cells and to enhance the sensitivity of a variety of malignant tumors to apoptosis induced by ionizing radiation and chemotherapeutic agents. However, the inherent limitations of E1A gene therapy prevent its application, such as the efficiency of expression, precision of targeting, and toxicity of vector. This prompted us to construct an E1A expression vector (pPIC9/E1A) and express the E1A protein in the methylotrophic yeast Pichia pastoris. The E1A protein was purified using two steps of ion-exchange column chromatography on HiTrap Q and HiTrap SP. The analysis indicated that the E1A protein/liposome inhibited S-180 tumor growth and also rendered the S-180 tumor strongly susceptible to the anticancer drug bleomycin in vivo. Furthermore, tunnel assay clearly revealed that the mechanism was induction of cellular apoptosis. Importantly, the E1A protein overcame the limitations of gene therapy. Thus the E1A protein may be a useful therapeutic agent for some malignant tumors. PMID- 12576671 TI - Apoptosis induced by influenza virus-hemagglutinin stimulation may be related to fluctuation of cellular oxidative condition. AB - Primary cultivated amnion epitherial cells prepared from amnion tissue of human fetal membrane (Amnion-cells) were stimulated with influenza virus-hemagglutinin (IV-HA), fractionated from a commercialized IV-HA vaccine by DEAE Sephacel column chromatography. From 72-96 h after stimulation, chromosomal DNA fragmentation and the appearance of in situ TUNEL stained-positive cells were revealed. Amnion-cell DNA fragmentation was inhibited in the presence of glycophorin A or C purified from the human erythrocyte membrane fraction, but not inhibited with free N acetyl-neuraminic acid. RT-PCR and Western blotting analysis showed that anti oxidative enzymes were altered with incubation period, accompanied by the expression of the cellular oxidative stress-related caspase cascade. Pre stimulation of Amnion-cells with hemin, a heme oxygenase-1 inducer, significantly attenuated IV-HA induced DNA fragmentation. It is concluded that IV-HA induces apoptosis in Amnion-cells, and that this apoptotic induction may be facilitated by certain sialoglycoproteins on the cell surface, and is related to changes in the intracellular redox condition. PMID- 12576672 TI - Calcineurin homologous protein isoform 2 (CHP2), Na+/H+ exchangers-binding protein, is expressed in intestinal epithelium. AB - The Na+/H+ exchangers (NHEs) comprise a family of membrane proteins that catalyze the electroneutral exchange of Na+ and H+. Calcineurin homologous protein (CHP) acts as a crucial cofactor for NHE activity through direct interaction with the carboxyl-terminal tail region of NHEs. We have cloned a new rat CHP isoform (rCHP2) and characterized the binding property to NHEs and the tissue distribution. rCHP2 binds to the juxtamembrane region of plasma membrane-type NHE isoforms (NHE1-5) in vivo and in vitro as well as rCHP1 (original rat CHP). Interestingly, CHP2 is predominantly expressed in the small and large intestine although rCHP1 shows relatively ubiquitous expression at both the mRNA and protein levels. In situ hybridization experiments demonstrated the abundant expression of CHP2 in the epithelial cell layer of villi of the small intestine in contrast with the expression of CHP1 in both the epithelial layer and connective tissues. These results suggest that CHP2 functions in the absorptive epithelium for the intestine with NHE(s). PMID- 12576673 TI - The ameliorating effects of acute and chronic administration of LiuWei Dihuang Wang on learning performance in rodents. AB - The ameliorating effects of LiuWei Dihuang Wang (LDW) after single, one-week or two-week treatment of scopolamine (SCOP)-induced and p-chloroamphetamine (PCA) induced amnesia by using the passive avoidance task and the facilitatory effects on two-way active avoidance performance in rats were studied. LDW (2 g/kg) after single treatment significantly prolonged the shortened step-through latency induced by SCOP and PCA. Then, SCOP- and PCA-induced amnesia was reversed by 1 and 0.1-1 g/kg LDW with one-week consecutive treatment respectively. For two-week consecutive treatment with LDW, the reversal from SCOP- and PCA-induced amnesia required only 0.01 g/kg. However, the rats treated with LDW only at 0.5, but not 0.01-0.1 g/kg, before or after each training session showed an increasing number of avoidances and a decreasing number of escapes on days 2-5 of learning. LDW at any dose alone did not influence the step-through latency in the training trial produced by non-shock rats, the motor activity and pentobarbital-induced hypnosis in rodents. These results suggest that LDW possesses the anti-amnestic and cognitive-enhancing activities related to the memory processes, and these activities were parallel to treatment duration and dependent on the learning models. PMID- 12576674 TI - Effects of microinjected carbachol on the antinociceptive response to noxious heat stimuli. AB - Injecting muscarinic receptor agonists into a specific area of the brainstem produces an antinociceptive response. The present study investigates whether direct injections of the cholinergic agonist, carbachol, into the rat nucleus reticularis gigantocellularis (NRGC)/nucleus reticularis gigantocellularis alpha (NRGCalpha) of the rostral ventrolateral medulla evokes antinociception, and then examines the interference action of cholinergic antagonists in rats. Microinjections of carbachol (0.75, 1.5, 3 micro g/site) prolonged hot plate (HP) and tail flick (TF) responses to noxious heat stimuli in a dose-dependent manner. The level of carbachol-induced antinociception during the HP and TF tests reached a maximum at 5-15 min after carbachol administration in all groups. Thereafter, the peak level progressively decreased and reached the baseline by the end of the experiment. Antinociception induced by carbachol at 3 micro g/site was attenuated by the prior administration of the muscarinic receptor antagonist, atropine (200, 500 ng/site). On the other hand, the nicotinic autonomic ganglion blocker, mecamylamine (1, 3 micro g/site), did not affect subsequent carbachol-induced antinociception. These results suggest that the antinociceptive effects induced by a microinjection of carbachol depend on muscarinic, but not nicotinic, mechanisms within the rat NRGC/NRGCalpha. PMID- 12576675 TI - Effects of Paeonia radix on 5-hydroxytryptamine synthesis and tryptophan hydroxylase expression in the dorsal raphe of exercised rats. AB - Paeonia radix is the root of Paeonia japonica MIYABE, a perennial plant classified in the family Paeoniaceae. In the present study, the effects of Paeonia radix on performance in treadmill exercise, and 5-hydroxytryptamine (5 HT) synthesis and tryptophan hydroxylase (TPH) expression in the dorsal raphe were investigated. Time to exhaustion in treadmill exercise was increased and exercise-induced increases in 5-HT synthesis and TPH expression in the dorsal raphe were shown to be suppressed by Paeonia radix treatment; 5-HT synthesis and TPH expression were inhibited by Paeonia radix treatment under resting conditions as well. In sum, treatment with Paeonia radix, inhibiting 5-HT synthesis and TPH expression, may bring about reduced fatigue, both during exercise and the resting state. PMID- 12576676 TI - Study of active substances involved in skin dysfunction induced by crowding stress. I. Effect of crowding and isolation on some physiological variables, skin function and skin blood perfusion in hairless mice. AB - The effects of five levels of population density on various organs, the neuroendocrine system, skin function, skin blood perfusion, and blood parameters were studied in the hairless mouse. Skin barrier recovery was evaluated by measuring transepidermal water loss after tape stripping. Blood perfusion was measured by means of a laser Doppler imaging technique. The effect of a parasympathetic nerve stimulator, carpronium chloride, on skin function in the crowded animal model was also examined. A 7 d crowding (10, 15, 20 mice/cage) significantly increased the levels of corticosterone, catecholamines (norepinephrine, epinephrine and dopamine), glucose and serum lactate dehydrogenase activity in circulating blood, induced atrophy of kidney, ovary and thymus and hypertrophy of adrenal glands, and decreased body weight gain in comparison with the control (5 mice/cage). Crowding also increased epidermal thickness and epidermal proliferative activity, and decreased corneocyte size, rate of barrier recovery and skin blood perfusion. Most of these changes became more marked with increasing population density and/or longer exposure to a crowded environment. Isolation (1 mouse/cage) increased the level of norepinephrine and rate of skin blood perfusion, and significantly delayed barrier recovery. Repeated topical applications of carpronium chloride for 7 d improved the changes in skin blood perfusion, barrier recovery, kidney and ovary, and epidermal morphology induced by crowding. The crowded animal model could be useful for quantifying objectively the influence of crowded environment-induced stress on cutaneous function and blood perfusion. PMID- 12576677 TI - Pharmacological evaluation of some new 2-substituted pyridine derivatives. AB - In the present study, a series of 2-substituted-pyridines were synthesized and characterized by IR, (1)H-NMR and Elemental Analysis. The compounds were assayed against seizures induced by maximal electro shock (MES) and pentylenetetrazole (scMet). Neurologic deficit was evaluated by the rotarod test. The decrease in the elevated motor activity by introceptive chemical stimuli (amphetamine antagonistic activity) was studied at the dose level of 25 and 50 mg/kg, antihistaminic and cardiac activity were also studied. All the compounds exhibited significant anticonvulsant activity. Compounds 2-(2'-piperazino ethanoxy)pyridine, 2-(3'-morpholino-2'-hydroxypropyloxy)pyridine, 2-(3' piperidino-2'-hydroxypropyloxy)pyridine and 2-(3'-piperazino-2' hydroxypropyloxy)pyridine were most active of the series against MES-induced seizures. Compounds 2-(2'-piperazino-ethanoxy)pyridine, 2-(2'-phenylamino ethanoxy)pyridine, 2-(3'-imidazolo-2'-hydroxypropyloxy)pyridine, 2-(3' methylamino-2'-hydroxypropyloxy)pyridine and 2-(3'-piperidino-2' hydroxypropyloxy)pyridine exhibited significant decrease in the elevated motor activity at the dose of 50 mg/kg. Remarkable sympathetic blocking activity was observed with 2-(3'-piperazino-2'-hydroxypropyloxy)pyridine, 2-(3'-piperidino-2' hydroxypropyloxy)pyridine and 2-(3'-imidazolo-2'-hydroxypropyloxy)pyridine only. Compounds 2-(2'-morpholino-ethanoxy)pyridine, 2-(2'-piperidino-ethanoxy)pyridine, 2-(2'-piperazino-ethanoxy)pyridine, 2-(2'-imidazolo-ethanoxy)pyridine, 2-(2' diphenylamino-ethanoxy)pyridine, 2-(2'-diethanolamino-ethanoxy)pyridine, 2-(2' phenylamino-ethanoxy)pyridine and 2-(2'-(4"-hydroxy)phenylamino-ethanoxy)pyridine exhibited significant blocking of histamine induced contraction on guinea pig ileum. PMID- 12576678 TI - Synthesis and biological activities of 2,6-diaryl-3-methyl-4-piperidone derivatives. AB - In the present study, a new series of 2,6-diaryl-3-methyl-4-piperidones was synthesized by Mannich reaction (condensation) of ethyl-methyl ketone, substituted aromatic aldehydes and ammonium acetate. Oximes and thiosemicarbazone derivatives of 2,6-diaryl-3-methyl-4-piperidones were synthesized by reaction with hydroxylamine hydrochloride and thiosemicarbazide respectively. The chemical structures were confirmed by means of IR, 1H-, 13C-NMR and mass spectral data. The compounds were screened for acute toxicity, analgesic, local anaesthetic and antifungal activity. 2-(4-Methylphenyl)-3-methyl-6-(4-chlorophenyl)-piperidin-4 one 2 exhibited the highest analgesic and local anaesthetic activity. The oximes and thiosemicarbazones were completely devoid of analgesic and local anaesthetic activity. 2-(4-Methylphenyl)-3-methyl-6-(4-hydroxyphenyl)-piperidin-4-oxime 21 and 2-(4-methoxyphenyl)-3-methyl-6-(4-chlorophenyl)-piperidin-4-oxime 17 exhibited potent antifungal activity against Aspergillus niger. Antifungal activity against Candida albicans was observed only with 2-(4 dimethylaminophenyl)-3-methyl-6-(4-chlorophenyl)-piperidin-4-oxime 20. 2,6-Diaryl 3-methyl-4-piperidones did not exhibit antifungal property. PMID- 12576679 TI - Effects of herbal drugs prescribed in wood creosote pills on the dissolution profile of guaiacol. AB - Wood creosote pills (P4) containing wood creosote and four herbal drugs, Gambir, Phellodendri Cortex, Glycyrrhizae Radix, and Citri Unshiu Pericarpium (CUP), have been used to treat food poisoning and diarrhea through self-medication in Japan. The mean dissolution time (MDT) of guaiacol, one of the active constituents of wood creosote, from P4 (138.3+/-3.3 min) was significantly longer than that (42.6+/-4.3 min) from pills (P0) containing only wood creosote. The MDT of the variant pills prepared from P4 without CUP (54.3+/-12.5 min) was found to be significantly shorter than that of P4. These findings suggest that CUP plays an important role in sustaining the dissolution of guaiacol from P4. The long MDT of guaiacol is considered one of the most important factors affecting the duration of efficacy after oral administration of wood creosote pills. The present findings are considered proof that CUP has been prescribed in traditional as well as new formulations of wood creosote pills. PMID- 12576680 TI - Effect of indinavir on the intestinal exsorption of amprenavir, saquinavir and nelfinavir after intravenous administration in rats. AB - To elucidate drug interaction between human immunodeficiency virus (HIV) protease inhibitors (PIs), the effect of indinavir (IDV) on the intestinal exsorption of other HIV PIs, amprenavir (APV), saquinavir (SQV) and nelfinavir (NFV) was investigated in rats using an in situ single perfusion method. IDV in the intestinal perfusate inhibited the exsorption of rhodamine 123 (Rho123), a known P-glycoprotein (P-gp) substrate, from blood into intestinal lumen in a concentration-dependent manner, and the inhibitory potency of 10 micro M IDV in the perfusate was close to that of 10 micro M cyclosporin A (CsA) in the perfusate. Ten micro M of IDV in the intestinal perfusate also decreased significantly the exsorption clearance of Rho123 after intravenous administration. The IDV concentration in this system was not likely to cause hepatic interaction between HIV PIs, because the plasma IDV concentration was far below its inhibition constants for other HIV PIs in the liver microsomes. Thus, 10 micro M of IDV was chosen to investigate the effect of this inhibition on the exsorption of APV, SQV and NFV. IDV in the intestinal perfusate markedly increased the exsorbed amounts of SQV and NFV but not APV after intravenous administrations. Their exsorption clearances, however, showed only a slight increasing tendency or remained unchanged. These findings suggest that in addition to P-gp inhibition, other factors such as CYP3A inhibition might be important in the drug interaction of IDV with APV, SQV and NFV after intravenous administration in rat small intestine. The results obtained in this study will provide useful information to discuss the interactions among PIs when a double protease therapy is used for in HIV-infected patients. PMID- 12576681 TI - Up-regulation of MDR1 function and expression by cisplatin in LLC-PK1 cells. AB - To examine whether cisplatin affects the multidrug transporter MDR1/P glycoprotein in the kidneys, the effects of cisplatin on cell sensitivity to an anticancer drug, MDR1 function and expression were examined by assessing the growth inhibition by the MDR1 substrate paclitaxel, the uptake and efflux of the MDR1 substrate Rhodamine123 and the level of MDR1 mRNA, respectively. Porcine kidney epithelial LLC-PK1 cells were used, as they have a structure and function similar to those of renal proximal tubular cells and physiologically express low levels of MDR1. The growth inhibitory curve of LLC-PK1 cells by paclitaxel was shifted to a higher concentration range by pretreatment with 1 micro M cisplatin for 48 h. The uptake and efflux of Rhodamine123 were significantly reduced and enhanced, respectively, by pretreatment with 1 micro M cisplatin for 48 h. This enhanced efflux was suppressed by the representative MDR1 substrate/inhibitor ciclosporin. The expression of MDR1 mRNA was increased by the existence of cisplatin for 48 h. These observations taken together suggested that the transient exposure to cisplatin could cause the up-regulation of MDR1 in LLC-PK1 cells. PMID- 12576682 TI - An approach to predict the ductus-arteriosus dilating effect induced by lipo prostaglandin E1 in newborn rats lacking plasma concentration-time data by the pharmacological response kinetic model. AB - The usefulness of kinetic analysis of pharmacological response data was discussed in investigating the ductus arteriosus dilating effect (DADE) of lipo-PGE1 (a lipid emulsion preparation of prostaglandin E1 for injection) preparations. Lipo PGE1 was administered intravenously via the umbilical vein by a bolus injection or an infusion in newborn rats 60 min after the delivery. The DADE data were expressed as the inner diameter ratio between the ductus arteriosus and the main pulmonary artery, and were analyzed by a pharmacological response kinetic (PRK) model consisting of an Emax model and a simple pharmacokinetic model as the pharmacodynamic- and the pharmacokinetic-component, respectively. The latter component includes the release process of free-PGE1 from the lipid phase of lipo PGE1, followed by distribution to the effect compartment. The Emax value was estimated by the maximal DADE observed 10 min after the bolus administration of each dose, and the value was fixed in the PRK analysis. The regression curves given by simultaneous non-linear least squares regression analysis were satisfactorily fitted to the observed DADE data at all doses. Prediction of the DADE of lipo-PGE1 in an infusion study was satisfactorily done using the estimated parameters in the i.v.-study. These findings indicate that PRK modeling based on the intensities of the observed pharmacological response-time data is a meaningful tool in some targeting-type drugs, for which pharmacokinetic analysis itself is meaningless or acquisition of pharmacokinetic data is technically impossible, in predicting the time courses of the drug's pharmacological response in different dosage regimens. PMID- 12576683 TI - Glucuronidation of propofol and its analogs by human and rat liver microsomes. AB - Propofol (2,6-diisopropylphenol), widely used an intravenous anesthetic, is rapidly metabolized to its glucuronide in the in vivo studies. Kinetic parameters for the glucuronidation of propofol and its analogs, such as 2,5 diisopropylphenol, 2-tert-butyl-6-methylphenol, 2-tert-butyl-5-methylphenol, 2,6 dimethylphenol and 2,5-dimethylphenol, were determined in vitro using human and rat liver microsomes. 2,5-Dimethylphenol and 2-tert-butyl-6-methylphenol exhibited the highest and lowest glucuronidation rates, respectively. Substitutes at the 2,6-positions gave lower glucuronidation rates than those at the 2,5 positions in both the human and rat microsomes. 2,5-Diisopropylphenol was glucuronidated at a lower rate in human than propofol. The affinity of uridine 5' diphosphate (UDP)-glucuronosyltransferase for disubstituted phenols, such as propofol, 2,5-diisopropylphenol, 2,5-dimethylphenol, and 2-tert-butyl-6 methylphenol, gave higher Km values in human liver microsomes than in rat ones, and lower Vmax values showed similar relationship, expect for Vmax in propofol. The alkyl group at the 6 position showed a higher Km for glucuronidation by a steric hindrance in the human and rat microsomes. Our results propose that the glucuronidation of propofol and its analogs may not be explained by only a steric hindrance. PMID- 12576684 TI - Oxide terpenes as human skin penetration enhancers of haloperidol from ethanol and propylene glycol and their modes of action on stratum corneum. AB - In this study, two terpenes with the same functional group; limonene oxide and pinene oxide were used at 5% w/v concentration in 50% v/v ethanol and 100% v/v propylene glycol (PG) to enhance the in vitro permeation of haloperidol (HP) through the human epidermis (or stratum corneum, SC). The enhancement mechanism of terpenes from both solvents was elucidated with HP-SC binding studies, Fourier transform infrared spectroscopy and differential scanning calorimetry. The enhancement activity of these terpenes was higher in 50% v/v ethanol than in 100% v/v PG. These terpenes in 50% v/v ethanol were predicted to provide the required therapeutic plasma concentration and daily-permeated amounts of the drug. Limonene oxide showed higher enhancement in both solvents, which was attributed to its less bulky structure. The terpenes in both solvents did not increase the partition of HP. Instrumental studies showed that these terpenes in 50% v/v ethanol extracted the SC lipids, disrupted the bilayer packing and partially fluidised the lipids. Limonene oxide in 100% v/v PG possibly disrupted the lipid bilayer, whilst leaving the overall bilayer structure intact and pinene oxide in the same vehicle fluidised the lipids within the ordered environment. This study showed that the mode of interactions of terpenes with SC were different in two solvent systems. PMID- 12576685 TI - Novel biological activity of the region (106-126) on human prion sequence. AB - We report that the synthetic peptide Prp106-126 (KTNMKHMAGAAAAGAVVGGLG-COOH) and the reversed peptide Prp126-106 (GLGGVVAGAAAAGAMHKMNTK-COOH) of human prion (hPrp) can express the decarboxylase activity for oxaloacetate in the presence of trifluoroethanol, similar to that of Oxaldie 1 (LAKLLKALAKLLKK-CONH2) reported previously. The degree of the relative activity of Prp106-126 and Prp126-106 to Oxaldie 1 is 0.47 and 0.21, respectively. Based on this experimental result, we applied the informational system method (ISM) developed by Veljkovic et al. to the amino acid sequence of Prp106-126 and Prp126-106 to extract a common factor. The same spectra were obtained, indicating that the same periodicity may be conserved on their sequences, as a necessary factor for expressing the same biological activity, irrespective of the orientation of the primary sequence. PMID- 12576686 TI - Effect of CAWS, a mannoprotein-beta-glucan complex of Candida albicans, on leukocyte, endothelial cell, and platelet functions in vitro. AB - Candida albicans is a medically important fungus which induces a disseminated candidasis and candidemia in immunocompromised hosts, and releases a polysaccharide fraction into the blood. We recently found that C. albicans released a water-soluble polysaccharide fraction (CAWS) into synthetic medium and demonstrated that CAWS was mainly composed of a complex of mannan and beta glucan. In the murine system, CAWS showed a lethality resembling anaphylactic shock when administered i.v., and induced coronary arteritis similar to Kawasaki Disease (KD) when given i.p. In the present study, we examined the biological activity of CAWS in the cell culture and found the following: i) CAWS slightly induced production of IFN-gamma and IL-6 by splenocytes at lower dose (ca. 10 micro g/ml), but at a higher dose strongly inhibited the proliferation of splenocytes induced by a B cell mitogen, lipopolysaccharide (LPS) and a T cell mitogen, concanavalin A. ii) The viability of these splenocytes monitored by propidium iodide staining was significantly reduced. iii) The addition of CAWS to a culture of monophage RAW264.7 cells significantly reduced cellular growth rate dose dependently. iv) The LPS-mediated synthesis of cytokines by RAW264.7 cells was significantly inhibited by CAWS. v) CAWS induced an aggregation of platelets in human platelet-rich plasma, and vi) CAWS inhibited the production of thrombomodulin by human umbilical endothelial cells and acted synergistically with TNF-alpha. Thus, CAWS strongly inhibited the cellular functions of leukocytes in vitro, partly through direct cytotoxicity. The enhanced production in injured cells of the vascular endothelium would be related to the local inflammatory response in the coronary artery. PMID- 12576687 TI - Effects of Bak Foong Pill on blood coagulation and platelet aggregation. AB - The effects of Bak Foong Pill (BFP, also known as Bai Feng Wan), a preparation of crude drugs in wide clinical use for treatment of gynecological disorders, on blood coagulation and platelet aggregation were investigated. The anticoagulant effect of BFP was evaluated by using thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (APTT) assays. Results showed that BFP 70% ethanol extract (BFP-E-ext) significantly prolonged the TT in a dose dependent manner with values of 17.6, 38.3, and 50.4 s at concentrations of 4.0, 6.0, and 12.0 mg/ml, respectively. Whereas, the BFP-E-ext did not show significant prolonging effect in PT and APTT assays. The results suggest that the anticoagulant effect of BFP is mediated by directly blocking thrombin, the key enzyme in the blood coagulation cascade. BFP-E-ext significantly inhibited platelet aggregation induced by collagen and adenosine diphosphate (ADP) with inhibition percentages of 74 and 52% at a concentration of 6.0 mg/ml, respectively, whereas, it exhibited a weak inhibitory activity on platelet aggregation induced by archidonic acid (AA). Comparing to BFP-E-ext, the effects of BFP aqueous extract (BFP-W-ext) on both anticoagulant and antiplatelet activities were significantly less potent. Moreover, the effects of the 26 ingredients of BFP on blood coagulation and platelet aggregation were separately evaluated with 19 ingredient herbs exhibiting anticoagulant effect and 10 exhibiting antiplatelet effect. The anticoagulant and antiplatelet effects of BFP were collectively demonstrated by in vivo assays showing prolonged bleeding times after BFP treatment for two weeks. The results of the present studies may provide explanations for beneficial effects of BFP on the circulation and indicate its potential use for cardiovascular diseases. PMID- 12576688 TI - Pharmacokinetic study of beta-methyldigoxin by enzyme immunoassay using a novel specific antiserum in rats. AB - We previously showed that enzyme immunoassay (EIA) of beta-methyldigoxin (MDx3) using anti-MDx3 3'-hemisuccinate-bovine serum albumin antiserum (Antiserum-I) was superior to that using commercial anti-digoxin antiserum (Antiserum-II) in terms of specificity and that pretreatment of human serum with phenyl boric acid (PBA) column was effective. In the present study, we examined the precision of EIA using Antiserum-I and the recovery of MDx3 after PBA column treatment in rat serum, and also investigated pharmacokinetic changes of MDx3 in rats. The intra- and inter-assay variations and recovery tests using Antiserum-I were good. The PBA column was effective in selectively separating MDx3 from rat serum containing MDx3 and its metabolites. The recovery tests using Antiserum-I with PBA column showed about 110% and the interference of metabolites of MDx3 was negligible. Serum concentration-time courses of MDx3 by EIA using Antiserum-I with PBA column and Antiserum-I were lower than that using Antiserum-II. The distribution volume at steady state and total body clearance values of MDx3 in these conditions were significantly higher than those using Antiserum-II. The usefulness of PBA column was ascertained, while effects of PBA column on these parameters were not significant. In addition, rapid absorption of MDx3 was observed by EIA using Antiserum-I with PBA column. These results suggest that EIA using Antiserum-I with PBA column for the pretreatment of serum samples should be a more useful and valuable system in therapeutic drug monitoring and pharmacokinetic studies of the unchanged type of MDx3 than Antiserum-II. PMID- 12576689 TI - Application of human Fc epsilon RI alpha-chain-transfected RBL-2H3 cells for estimation of active serum IgE. AB - We transfected the alpha-chain of human FcepsilonRI into rat basophilic leukemia cell line RBL-2H3, established several stable transfected cells, and screened them by beta-hexosaminidase release induced by sensitization with human IgE and stimulation with anti-human IgE antibody. A cloned cell line RBL-hEIa-2B12 was the strongest responder among the transfected cell clones. The concentrations of cytosolic free Ca2+ concentration in the human IgE-sensitized cells increased after stimulation with anti-human IgE antibody. Thus, it is suggested that the alpha-chain of human FcepsilonRI is associated with the beta-chain and/or gamma chain of rat FcepsilonRI, and that they form functional high affinity IgE receptor complexes. The total IgE concentrations of the sera from allergic patients were determined by using the beta-hexosaminidase release assay, where the transfected cells were sensitized with diluted and heat-inactivated (at 56 degrees C for 30 min) serum and stimulated with anti-human IgE antibody. The IgE concentration obtained correlated with those measured by an enzyme immunoassay method. beta-Hexosaminidase release induced by stimulation with 5 times diluted serum was sometimes less than the release induced by the same serum; diluted 25 times or 125 times, suggesting that these serum contained factors that blocked IgE binding to FcepsilonRI or cross-linking by anti-human IgE antibody. The results suggested that our system will be useful for detecting FcepsilonRIalpha bindable IgE in human serum. PMID- 12576690 TI - Characterization of calreticulin as a phosphoprotein interacting with cold induced protein kinase in rice. AB - Calreticulin is an abundant endo/sarcoplasmic reticulum Ca2+-binding protein. To investigate whether calreticulin (CRO1) is involved in the cold-stress response in rice, a transgenic plant was constructed. The transcriptional level was decreased within 30 min and recovered within 2 h of a cold treatment. The calreticulin protein was shifted from a soluble fraction to an insoluble fraction by cold stress. Endogenous abscisic acid (ABA) is an important factor in cold response, and the synthesis of ABA was strongly induced in CRO1-sense transgenic rice, the same as in cold-sensitive rice. The phosphorylation of calreticulin increased after cold treatment. Over-expression of calreticulin enhanced the activities of 47 kDa Ca2+-dependent protein kinase (CDPK) that had been induced by cold treatment. The 47-kDa CDPK activity increases more in the cold sensitive variety IR36 and the sense transgenic rice than it does in other varieties. The synthesis of ABA, phosphorylation of calreticulin and 47-kDa CDPK activity induced in sense transgenic rice were the same as in cold-sensitive rice and the phosphorylation of antisense transgenic rice was similar to that of cold-tolerant rice. These results suggest that the calreticulin is involved in the signaling pathway leading to response to cold stress. PMID- 12576691 TI - Molecular cloning of the gene for edeine B1 amidinohydrolase in addition to the agmatinase activity in Bacillus subtilis. AB - A gene with a high-nucleotide sequence homology to the edeine B1 amidinohydrolase gene of Bacillus brevis was identified in the database of the Bacillus subtilis genome. The gene was isolated, expressed in Escherichia coli, and the gene product was analyzed with regard to the characteristics of its enzyme activity. A 32-kDa protein encoded by the ywhG gene showed a 69.8% amino acid sequence homology to the edeine B1 amidinohydrolase of B. brevis. Among various guanidino compounds, edeine B1 and agmatine were both efficiently hydrolyzed by the protein encoded by the ywhG gene, although edeine B1 was a more potent substrate than agmatine in this assay system. These data indicate that the protein encoded by the ywhG gene is an agmatinase that is essential for polyamine biosynthesis in B. subtilis. PMID- 12576692 TI - Functional cloning and expression of emeA, and characterization of EmeA, a multidrug efflux pump from Enterococcus faecalis. AB - A fragment of chromosomal DNA from Enterococcus faecalis ATCC 29212 was cloned using Escherichia coli KAM32 host cells lacking major multidrug efflux pumps. E. coli KAM32 cells were sensitive to many antimicrobial agents, and the transformed cells harboring a recombinant plasmid became resistant to several structurally unrelated antimicrobial agents such as tetraphenylphosphonium chloride, 4',6 diamidino-2-phenylindole (DAPI), Hoechst 33342, acriflavine, benzalkonium chloride, norfloxacin and ethidium bromide. This suggests that the cloned DNA fragment carries a gene(s) encoding a multidrug efflux pump. Determination of the nucleotide sequence of the cloned DNA revealed a gene designated as emeA. The transformed E. coli cells showed efflux activity of several antimicrobial agents such as DAPI, Hoechst 33342 and acriflavine. Efflux of DAPI via EmeA was strongly inhibited by reserpine. PMID- 12576693 TI - Anti-tumor promoting effect of glycosides from Prunus persica seeds. AB - Four minor components, along with the major cyanogenic glycosides, amygdalin and prunasin, were isolated from Prunus persica seeds (Persicae Semen; Tounin), and characterized as mandelic acid glycosides (beta-gentiobioside and beta-D glucoside) and benzyl alcohol glycosides (beta-gentiobioside and beta-D glucoside). The anti-tumor promoting activity of these compounds was examined in both in vitro and in vivo assays. All of the compounds significantly inhibited the Epstein-Barr virus early antigen activation induced by tumor promoter. In addition, they produced a delay of two-stage carcinogenesis on mouse skin that was comparable in potency to (-)-epigallocatechin gallate from green tea. Structure-activity relationships indicated that a substituent at the benzylic position with glycosidic linkage affected the in vitro and in vivo activities with an order of enhancing potency, CN or =2.5 cm in diameter, persisting for more than 7-14 days, without a corpus luteum (CL) were used for the study. PRID or placebos were inserted into the vagina for 12 days. Five animals lost the intravaginal device before removal and one was culled. Based on plasma progesterone concentration on the day of treatment, 20 (17 PRID and 3 placebos) of the remaining 34 cows had follicular cysts (progesterone < or =1 ng/ml) and 14 (10 PRID and 4 placebos) had luteal cysts (progesterone >1 ng/m l). Fourteen (82%) of the PRID-treated follicular cystic cows responded with formation of a CL within 14 days after treatment, and an overall conception rate of 53.8%. Likewise, 70% of the treated luteal cystic cows responded with CL formation and 71.4% conception rate. No significant differences were observed in hematocrit (Ht), white blood cell count and serum levels of glucose, blood urea nitrogen, aspartate aminotransferase, and alanine aminotransferase, between the day of PRID insertion and removal, in animals with follicular and luteal cysts. PRID treatment resulted in ovulation 2-4 days later and formation of a CL in cows that recovered. PMID- 12576706 TI - A study on the number of recovered spermatozoa in the uterine horns and oviducts of gilts, after fractionated or non-fractionated insemination. AB - The objective of this study was to compare the number of recovered spermatozoa, in different parts of the uterine horn and oviduct in gilts, after insemination with fractionated (experiment) and non-fractionated (control) liquid stored semen. The number of spermatozoa and volume of backflow was also investigated. Twenty three cross-bred gilts were used in the study. They were divided into 2 groups, a control group (non-fractionated liquid stored semen, n=10) which were inseminated with 100 ml of liquid stored semen containing 3,000 million spermatozoa per dose and an experimental group (fractionated liquid stored semen, n=10) which were inseminated with 50 ml of liquid stored semen, with 3,000 million spermatozoa per dose and followed by another 50 ml of semen dilutor (Beltsville Thawing Solution, BTS). Thereafter, backflow semen was collected and measured every 15 min for a period of 1 hr. Three or 12 hr after insemination, 5 gilts from each group had the uterus, the horn of the uterus, the oviducts and the ovaries removed under general anaesthesia. The horn of uterus and the oviducts were seperated by ligation into 6 segments. All 6 segments were flushed with BTS to collect all spermatozoa within the segment. Recovered spermatozoa were counted, using a haemocytometer and the volume recorded. It was seen that the percentage of spermatozoa in the backflow semen in the experimental group was less than in the control group. The difference was not significant in the gilts that were operated on 3 hr after insemination, the mean number of spermatozoa in the uterine horn and the utero-tubular junction (UTJ) was more in the experimental than in the control group, but less in the isthmus and the ampulla of the oviduct. The gilts which were operated on 12 hr after insemination, had relativity more ovulating gilts in the control group than in the experimental group (3 of 4 gilts compare to 3 of 5 gilts). The control group had more spermatozoa in the oviduct than the experimental group, but less in UTJ and in the horn of the uterus. Again the difference was not significant. It can be concluded that fractionated (experimental) or non-fractionated (control) insemination of semen with the same number of spermatozoa provides no significant difference in the number of spermatozoa either in the horn of the uterus, the UTJ or the oviduct of gilts. PMID- 12576707 TI - Protective effects of oral microencapsulated Mycoplasma hyopneumoniae vaccine prepared by co-spray drying method. AB - The efficacy of Mycoplasma hyopneumoniae oral vaccine was investigated in microsphere dosage form. A co-spray drying process was used to apply an encapsulating material, Eudragit L30 D-55, to microspheres containing Mycoplasma hyopneumoniae antigens. The microspheres were generally effective (>93%) with protein release at pH 7.4, but almost none were released at pH 1.2, for 3 hr in an in vitro dissolution test. An SPF-swine model was used to evaluate the effectiveness of the microspheres as an oral vaccine, and the related immune responses. The serum's systemic IgG against M. hyopneumoniae was evoked by ELISA analysis, after a 2nd immunization of all pigs. The vaccinated groups' mean lesion score was significantly lower after the Mycoplasma hyopneumoniae challenge than that of the nonvaccinated/challenged groups (P<0.05). This study strongly suggests that the oral microspheres vaccine prepared by a co-spray drying method can provide effective protection against M. hyopneumoniae infection in pigs. PMID- 12576708 TI - Amino acid polymorphisms of PrP gene in Mongolian sheep. AB - To characterize amino acid polymorphisms in sheep prion protein (PrP), we analyzed the PrP genes from 271 sheep of 4 breeds (Khalkh, Yeroo, Orkhon and Khangai) raised in central Mongolia (Tuv, Uvurkhangai and Selenge prefectures). A total of 16 genotypes and 8 allelic variants of the PrP gene at codons 112, 136, 154 and 171 were found. At codon 171, 1.8% of the sheep had arginine/arginine (R/R) (resistant to scrapie) and 66.8% had glutamine/glutamine (Q/Q) (susceptible to scrapie). Several Yeroo and Orkhon sheep raised in Selenge prefecture had valine at codon 136 (136V) (highly susceptible to scrapie). Several Yeroo, Orkhon and Khangai sheep raised in Selenge prefecture had histidine at codon 154 (154H). Novel polymorphisms of valine (V) and serine (S) at codon 127, lysine (K) at codon 171, and leucine (L) and arginine (R) at codon 189 were also found in Khalkh, Yeroo and Orkhon sheep. It is not known whether these novel polymorphisms affect scrapie susceptibility. PMID- 12576709 TI - Effects of DDAVP administrated subcutaneously in dogs with aspirin-induced platelet dysfunction and hemostatic impairment due to chronic liver diseases. AB - To evaluate the hemostatic effects of desmopressin (DDAVP) in dogs with aspirin induced platelet dysfunction and hemostatic impairment in chronic liver diseases, 3 microg/kg DDAVP was administrated subcutaneously. In aspirin-induced platelet dysfunction dogs (n=5), prolonged BMBT (buccal mucosal bleeding time) was shortened significantly after DDAVP injection (2.2 +/- 1.2 min, P<0.05). In dogs with chronic liver diseases (n=4), activated partial thromboplastin time (APTT) tended to shorten by 0.9 to 3.0 sec, and prolonged BMBT was shortened in two cases for 4.2 and 1.7 min after DDAVP injection. Therefore, the present results indicated that DDAVP shortened the prolonged BMBT in dogs with aspirin-induced platelet dysfunction and chronic liver disease. DDAVP might be helpful in hemostasis under invasive procedures such as biopsy or surgery for dogs with hemostatic impairment. PMID- 12576710 TI - Latency and persistence of bovine herpesvirus type 4, strain B11-41, in bovine nervous tissues. AB - Three cattle were experimentally infected with bovine herpesvirus type 4 (BoHV 4), strain B11-41, isolated from the spinal cord of a cow, and monitored for clinical symptoms. None of them showed any clinical signs except increases of leukocyte numbers in two of them, and the body temperature remained normal throughout the experiment. Antibody titers against BoHV-4 continuously increased for one month and were maintained at a high level for more than 1 year by enzyme linked immunosorbent assay (ELISA). The virus was isolated only from serum and peripheral blood leukocytes (PBL) of one cow in the early stage of infection, but the viral genome was detected in PBL continuously by PCR. When they were euthanized, the viral genome was detected in the lymph nodes and nervous tissues such as medulla, spinal cord, and trigeminal ganglion. These results indicate that cattle are infected with the virus latently and persistently, and the latency site would be in the tissues of the central nervous system as well as lymphoid tissues. When a seroepidemiological survey was performed on antibodies to BoHV-4 among cattle in Japan by ELISA, the rate of antibody-positive cattle was 8.9% and they were found irregularly on certain farms. PMID- 12576711 TI - Hemagglutination and antigenic comparison of rabbit hemorrhagic disease virus. AB - The hemagglutinating activity and serological properties of three strains of rabbit hemorrhagic disease virus, Chinese, Korean and Shizuoka, which was first isolated in Japan, were examined by hemagglutination (HA) and cross hemagglutination inhibition (HI) test with human erythrocytes. Similar results were observed between the Chinese and Korean strains, both of which gave positive HA at 4 degrees C with O, A, B and AB, and at 22 degrees C with B and AB blood groups. In the Shizuoka strain, positive HA was observed at 4 degrees C with O, A, B and AB, at 22 degrees C with A, B And AB, and at 37 degrees C with B blood group. In experimentally infected rabbits, HI antibody in these animals showed a titer of 16,384 or 32,768 at 4 weeks after inoculation. No serological difference was observed in three strains by cross HI test. PMID- 12576712 TI - Changes in sexual behavior and fecal steroid hormone concentrations during the breeding season in female Hokkaido brown bears (Ursus arctos yesoensis) under captive condition. AB - The longitudinal changes in fecal steroid hormone concentrations and sexual behavior in 2 mated/pregnant and 3 non-mated female Hokkaido brown bears were investigated during the breeding season. Behavioral estrus (standing) lasted for 14 and 32 days in the mated females and for 25 to 36 days in the non-mated females. In non-mated females, sexual behavior, such as female-female mounting and masturbation, was observed for several days before and after the estrous period. In mated females, mean fecal estradiol-17beta concentrations were higher in the estrous period than in the post-estrous period, while fecal progesterone concentrations were higher in the post-estrous period than in the estrous period. The similar trends of steroid hormone changes were observed in the non-mated females. PMID- 12576713 TI - Changes of biomechanical characteristics of the bone in experimental tibial osteotomy model in the dog. AB - Experimental osteotomy model of canine tibia was prepared to investigate the changes in biomechanical characteristics during the healing process. After 16 weeks, although radiographs revealed that fracture healing proceeded, the recovery rates of the mechanical parameters ranged from 12.3 to 47.3%, compared to the intact side. After 32 weeks, those recovered to 68.9-93.2%. These results suggested that the biomechanical characteristics of the healing bone could not be recovered sufficiently even after the passage of the healing period which has been empirically proposed from clinical findings. It was also considered that evaluation of the mechanical parameters using this osteotomy model would be suitable for investigating the effect of osteoinductive growth factors on fracture healing. PMID- 12576714 TI - Cytotoxic T-lymphocyte activity specific for hemagglutinin (H) protein of canine distemper virus in dogs. AB - Cytotoxic T-lymphocyte (CTL) responses to hemagglutinin (H) protein of canine distemper virus (CDV) were evaluated in dogs using the replication-deficient adenovirus protein expression system. Skin fibroblasts were isolated from two dogs and were infected with recombinant adenovirus bearing the CDV-H gene (Ade CDVH). CTL assay was performed using fibroblasts expressing CDV-H protein as target cells and peripheral blood lymphocytes (PBL) collected from the same dogs one week after immunization of CDV as effector cells. Specific cytotoxic activity was observed against autologous but not heterologous fibroblasts expressing CDV-H protein. These results indicate that the CTL epitope(s) were localized in the H protein. PMID- 12576715 TI - Ganglioglioma in the thalamus of a puppy. AB - A solitary brain mass of a 4-month-old miniature dachshund showing seizure-like neurological signs was examined histopathologically. At necropsy a white tumor mass, replacing the thalamus, approximately 1.5 cm in diameter, was found. There was cystic space filled with yellowish pale fluid in the central area of the tumor mass. Histopathological examination revealed that the mass consisted of irregularly arranged well-differentiated neuronal and glial cells, and multifocal mineral deposits. The neuronal cells had a large clear nucleus and various amount of Nissl substances in the cytoplasm. Some neural cells were bi-nucleated. Neither mitotic figures nor proliferating cell nuclear antigen (PCNA)-positive nuclei was found in the neuronal cells. Immunostaining for glial fibrillary acidic protein (GFAP) revealed diffuse proliferation of GFAP-positive glial cells and their processes, while these glial cells did not show apparent cellular atypism, mitotic activity, or PCNA-immunoreactivity. Accordingly, the present tumor was diagnosed as ganglioglioma, and hamartomatous histogenesis might be possible. PMID- 12576716 TI - Adjuvant effects of sugar cane extracts (SCE) in chickens. AB - The effects of sugar cane extracts (SCE) on immune responses in chickens were studied. Two- or 10-month-old chickens orally administered SCE (500 mg/kg/day), for 3 consecutive days before immunized with sheep red blood cells, Brucella abortus and Salmonella Enteritidis organisms, showed significantly increased and prolonged antibody responses to these antigens, compared to control chickens without SCE. Furthermore, chickens orally administered SCE also revealed enhanced delayed type hypersensitivity responses to human gamma globulin. These results indicated that SCE has immunostimulating and adjuvant effects in chickens. PMID- 12576717 TI - Seasonal change in the number of Cryptosporidium parvum oocysts in water samples from the rivers in Hokkaido, Japan, detected by the ferric sulfate flocculation method. AB - An epidemiological study was carried out in natural water supplies of Hokkaido, one of the largest dairy prefectures in Japan. To investigate the prevalence of Cryptosporidium parvum (C. parvum) oocysts water samples were collected from three rivers in the eastern area of Hokkaido from August 1999 to October 2001, and C. parvum oocysts were collected and purified by the ferric sulfate flocculation method. The oocysts were detected using the immunofluorescent assay test (IFAT) and 4', 6-diamidino-2-phenylindole (DAPI) staining. The seasonal change in the number of oocysts detected was observed. Oocysts increased in numbers from the late summer to the early autumn (from August to November), thereafter, they exhibited a trend to decrease until December, when no oocysts could be detected. The maximum number of oocysts detected in the three rivers was 3.50, 5.00 and 3.33 oocysts/l, respectively. The oocyst density in river water changed in relation to the season in 1999, 2000 and 2001. This report first cleared up the seasonal changes in C. parvum oocysts number in river water. PMID- 12576718 TI - Cryptosporidium infection of cattle in the Tokachi district, Hokkaido. AB - The prevalence of Cryptosporidium infection was examined in 480 healthy cattle (0 39 months old) in the Tokachi district in Hokkaido during the period from June to September in 2000 and from June to July in 2001. C. parvum oocysts were detected in 6 of 50 cattle (0-2 months old) in 2001; while C. muris was detected in 2 of 56 cattle (6-8 months old) in 2001, in 1 of 15 cattle (9-11 months old) in 2001, in 1 of 88 cattle (15-17 months old) in 2000, in 4 of 89 cattle (18-21 months old) in 2000 and in 2 of 53 cattle (21-23 months old) in 2000. PMID- 12576719 TI - Characterization of Campylobacter lanienae from pig feces. AB - To isolate Campylobacter spp., the feces of healthy cattle, pigs, and broilers were examined between June 1999 and January 2000. Campylobacter lanienae strains were isolated from the feces of healthy pigs, but not from the feces of cattle or broilers. In six C. lanienae isolates, there was only 21-38% DNA-DNA homology to Campylobacter hyointestinalis subsp. lawsonii strain NCTC 12901. Thus, the primary host of C. lanienae is likely to be the pig and C. lanienae appears to be a species distinct from C. hyointestinalis subsp. lawsonii. In addition, an intervening sequence of 226 bp in the 16S rRNA gene was found in four isolates. PMID- 12576720 TI - Survey of benign Theileria parasites of cattle and buffaloes in Thailand using allele-specific polymerase chain reaction of major piroplasm surface protein gene. AB - During a year from 1999 to 2000, a total of 247 blood samples were collected from 214 cattle and 33 water buffaloes in 16 distinct geographical locations of Thailand and analyzed by allele-specific PCR amplification of major piroplasm surface protein (MPSP) genes of benign Theileria parasites. Four allelic MPSP gene types were determined namely C-type, I-type, B-type and Thai-type, which were originally designated from Japanese Theileria orientalis (Chitose, Ikeda), Australian T. buffeli (Warwick) and Thai T. sp. (Kamphaeng Saen), respectively. Only two allelic MPSP gene types were successively amplified from 204 (82.6%) blood samples. Among positive cases, 138 (67.6%) and 17 (8.3%) samples contained either Thai-type or C-type parasites, respectively, while 49 (24%) samples contained both types. However, nucleotide sequences of MPSP genes of Thai T. sp. amplified by C-type specific primers revealed higher (96.3%) similarity to Indonesian T. sp. rather than (87.8% similarity) to Japanese T. orientalis (Chitose) designated as C-type. PMID- 12576721 TI - Effects of salivary gland extract from Rhipicephalus sanguineus on IgG subclass production and cytokine mRNA expression in mononuclear cells of canine peripheral blood. AB - The effects of salivary gland extract (SGE) from R. sanguineus were examined on the production of IgG1 and IgG2 and the mRNA expression of IFN-gamma, IL-2, IL-4, IL-5 and IL-10 in the mononuclear cells from canine peripheral blood, treated with concanavalin A (ConA) in vitro. SGE suppressed the ConA-induced production of IgG2. It also inhibited the expression of IFN-gamma, IL-2 and IL-5 mRNA in a dose-dependent manner. No dose-dependent suppression was observed of IL-10 mRNA expression although a significant effect was observed at a SGE protein concentration of 25 microg/ml. SGE had no effect on the mRNA expression of IL-4. These results suggest that the suppression of IgG2 production by SGE from R. sanguineus was caused by the suppression of IFN-gamma production. PMID- 12576722 TI - Epidemiological investigations of an outbreak of intestinal atresia in two Israeli dairy herds. AB - An epidemiological investigation of an outbreak of intestinal atresia in Israeli Holstein-Friesian newborn calves showed a linkage with rectal palpation for early pregnancy diagnosis, performed less than 42 days after insemination. The odds of an exposed calf, i.e., one born to a dam that was diagnosed by early palpation as having intestinal atresia were 119.7 times higher than one born in normal control herds (95% CI; 7.4-1946.3). A total of 682 calves-at-risk was recorded from mid 1998 to mid-2000 and a total of 47 calves (6.9%) were born with intestinal atresia during this period. Two forms of intestinal atresia were recognized at post-mortem: atresia coli and atresia ilei. PMID- 12576723 TI - Effect of electroacupuncture on minimum alveolar concentration of isoflurane in dogs. AB - The effect of electroacupuncture (EA) on minimum alveolar concentration (MAC) of isoflurane was evaluated in dogs. After determination of baseline MAC, EA was applied at each acupoints (LI-4, SP-6, ST-36 and TH-8) and nonacupoint for 30 min. MAC was determined again. EA at acupoints significantly lowered the MAC of isoflurane in dogs (17.5 +/- 3.1%, 21.3 +/- 8.0%, 21.2 +/- 7.5% and 15.4 +/- 3.1%, respectively). In control group and nonacupoint electrical stimulation group MAC were not decreased significantly. From these results, electroacupuncture at each acupoints used in the present study would have an advantage in isoflurane anesthesia with reducing its requirement. PMID- 12576725 TI - Increased locomotor activity, increased food and water intake and decreased PVN neurons in H1 calponin gene-deficient mice. AB - Calponin (h1 or basic) is an actin-binding protein that is expressed abundantly in smooth muscle. Our previous study using h1 calponin-null mutant mice demonstrated that h1 calponin inhibits the shortening velocity of smooth muscle contraction without significantly affecting the amplitude of force production. Furthermore, early onset of osteogenesis and increased bone formation have been reported in mutated mice. In the present study, we examined the effect of h1 calponin depletion on the metabolism and behavior of mice and found that the mutated mice showed increased locomotor activity, as well as increased intake of food and water, associated with the decreased number of neurons in the paraventricular nucleus of the hypothalamus (PVN). PMID- 12576724 TI - Canine Bcl-xL gene and its expression in tumor cell lines. AB - The canine Bcl-xL gene was cloned and sequenced. Canine Bcl-xL cDNA clone was 1252 bp in length, and encoded 233 deduced amino acides. The predicted canine Bcl xL amino acid sequence shared 99.6%, 97.0%, 97.9%, 98.7% and 98.3% homology with that of human, mouse, rat, sheep and pig Bcl-xL, respectively. RT-PCR analysis revealed that canine Bcl-xL mRNA was constitutively expressed in CL-1 (canine lymphoma) and GL-1 (canine B cell leukemia) cell lines. PMID- 12576726 TI - Phenotypic analysis of hepatic lymphocytes from healthy dogs. AB - Phenotypes of lymphocytes from laparoscopically biopsied liver tissues of eleven healthy beagle dogs were analyzed. The proportion of CD3(+) lymphocytes (T cells), CD3 (-)CD21(+) lymphocytes (B cells) and CD3 (-)CD21(-) lymphocytes (non T non-B lymphocytes), and the CD4(+)/CD8(+) ratio in the canine hepatic lymphocytes were 54.8 +/- 11.9%, 4.7 +/- 3.1%, 40.7 +/- 13.2%, and 0.33 +/- 0.12, respectively, while those in peripheral blood lymphocytes were 85.4 +/- 6.5%, 9.3 +/- 6.1%, 5.3 +/- 1.8%, and 1.64 +/- 0.36, respectively. These results indicated that the constitution of hepatic lymphocytes quite differed from that of peripheral blood lymphocytes in dogs, and suggested that the regional immunity in canine liver might be specific. PMID- 12576728 TI - Inhalant allergies in Zimbabwe: a common problem. AB - Allergic rhinitis, allergic conjunctivitis, allergic asthma and atopic dermatitis affect a quarter of the population of the industrialised countries and are the most common symptoms of type I hypersensitivity reactions. Their prevalence has been documented in many communities, yet data from Africa are limited. In the 5 year period from September 1997 to September 2002, approximately 14,000 patients of all ages were referred to the only specialist allergy clinic in Harare, Zimbabwe, for allergy investigation, diagnosis and therapeutic management. In approximately one tenth of these patients, food allergies were diagnosed; less frequent presentations included allergic pharyngitis, bee- or wasp-induced reactions and latex allergy. An allergologic basis for the clinical conditions was established following a careful clinical and physical examination, evaluation of family history and the detection of in vivo (skin prick test) or in vitro (radio-allergo-sorbent test; RAST) allergen-specific IgE antibodies to known sources of inhalant allergens, using commercial allergen extracts and a RAST assay kit. Asthma diagnosis was supported by the clinical and laboratory findings of salbutamol-reversible airways obstruction. Case files of the first 1,046 patients are reviewed in order to define the clinical presentation of allergic patients in this region. It is highlighted that allergy in general and inhalant allergy in particular are very common, if not widely acknowledged, clinical problems in this Central African region. PMID- 12576729 TI - Three-dimensional structure of the panallergen Phl p 7. PMID- 12576730 TI - Cloning and molecular characterization of a cDNA from Blomia tropicalis homologous to dust mite group 3 allergens (trypsin-like proteases). AB - BACKGROUND: Exposure to the house dust mite Blomia tropicalis is increasingly being implicated as a major risk factor for asthma exacerbations in sensitized individuals. The objective of this study is to clone and characterize B. Tropicalis allergens in order to better define their role in allergic asthma. METHODS: A lambda gt22A cDNA library was constructed from B. Tropicalis mRNA and screened using specific DNA probes. A full-length cDNA (Bt2-3) was isolated, subcloned, and sequenced. RESULTS: Sequence analysis showed that clone Bt2-3 encodes the full-length serine protease preproenzyme from B. Tropicalis. The predicted Bt2-3 protein consists of a 15-amino-acid signal peptide, a 20-amino acid propeptide and a mature protein of 231 amino residues. BLAST analysis of the deduced amino acid sequence showed high homology (48-54%) between clone Bt2-3 and serine proteases (group 3 allergens) from domestic dust mites. Both the serine active site (DACQGDSGGPVA; amino acids 214-225) and the histidine active site (LTAAHC; amino acids 71-76) of serine proteases were highly conserved. The estimated molecular weight of the preproenzyme is 27.5 kD and its theoretical pI is 8.7. The mature protein contains a putative tyrosine kinase phosphorylation site (amino acids 24-31) and several N-myristoylation sites. Sequence alignment shows that the serine protease active sites are highly conserved among the clinically important mite species, including B. Tropicalis. CONCLUSION: We report the cloning and molecular characterization of a B. Tropicalis cDNA clone encoding a trypsin-like protease, with a possible major role as an allergen. Expression and further characterization of the recombinant product will help determine the role of proteolytic enzymes in the pathophysiology of allergic asthma. PMID- 12576731 TI - Gelatin prepared from tuna skin: a risk factor for fish allergy or sensitization? AB - BACKGROUND: Although fish gelatin may represent a useful alternative to bovine or porcine gelatin, the clearly recognized high prevalence of fish allergy could increase the risk of anaphylaxis to gelatin. The rationale for investigating tuna gelatin rather than gelatin from more allergenic fishes is the availability of an industrial gelatin under development. The infrequent occurrence of tuna allergy should influence the safety of a derived product. The present study investigated IgE antibodies to tuna-skin-derived gelatin in adults and children with documented fish allergy or sensitization. METHODS: Serum samples were taken from 100 consecutive patients with fish allergy or sensitization and tested for IgE antibodies against hydrolyzed or nonhydrolyzed gelatin extracted from tuna skin as compared to extracts from tuna flesh, tuna skin as well as bovine or porcine gelatin. Patients with tuna allergies or sensitization were sensitive to the same tuna species (yellowfin) as that from which the gelatin was obtained. IgE antibodies to these various extracts were analyzed using SDS-PAGE and immunoblotting. RESULTS: Only 3 of the 100 serum samples tested gave evidence of reactivity to gelatin extracted from tuna skin. Cross-reactivity between bovine/porcine and fish gelatin was not observed. CONCLUSION: The risk of adverse reactions to tuna skin gelatin seems to be significantly lower than the risk of fish allergy. Fish gelatin may represent a valuable alternative to bovine or porcine gelatin. PMID- 12576733 TI - Flow-cytometric cellular allergen stimulation test in latex allergy. AB - BACKGROUND: The use of flow-cytometric basophil activation to different allergens has been recommended in recent years. In this study, we analyzed the diagnostic reliability of the flow-cytometric allergen stimulation test (FAST) after latex specific stimulation in vitro. The diagnostic reliability of the technique was assessed as well as its correlation with other in vitro diagnostic parameters. METHODS: 43 patients allergic to latex with a positive history and skin test participated in the study. Thirty subjects (20 of them exposed to latex) with a negative history, skin tests and serum-specific IgE determination to latex were used as controls. In FAST the percentage of basophils that express CD63 as an activation marker after in vitro stimulation with allergen (latex) is determined by flow cytometry, following double labelling with the monoclonal antibodies anti CD63-PE and anti-IgE FITC. RESULTS: Intraclass correlation coefficient in FAST with latex was 0.995 (p < 0.0001), which demonstrates the excellent reproducibility of this technique. Taking a cutoff point of 10% by means of ROC curves, FAST yields a sensitivity of 93% and a specificity of 100%. The FAST positive predictive value in latex allergy was 100% and the negative predictive value was 99.9%. We found a positive and significant correlation between FAST and specific IgE (CAP) with the histamine release test and specific sulphidoleukotriene production [cellular allergen stimulation test (CAST); p < 0.05]. CONCLUSIONS: FAST is a highly reliable technique (93% sensitivity and 100% specificity) in the in vitro diagnosis of IgE-mediated latex allergy. PMID- 12576732 TI - Dose-dependent and preterm- accentuated diaplacental transport of nutritive allergens in vitro. AB - OBJECTIVE: The mechanisms by which nutritive allergens are transported from mother to fetus and the ensuing immunological response are incompletely understood. We investigated the role of different allergen concentrations in influencing the diaplacental allergen transport in preterm and term placentas. METHOD: Twenty-seven human term placentas and 12 preterm placentas were dually perfused in vitro for up to 4 h by adding alternately two different nutritive allergens, beta-lactoglobulin (BLG) or ovalbumin (OVA), at four different allergen concentrations (0.02, 0.2, 2 and 20 mg/ml) to the maternal perfusate medium. Allergen concentrations in fetal venous outflow samples collected during perfusion were measured by using specific ELISAs. RESULTS: Perfusion of increasing allergen concentrations via the maternal circulation resulted in a concentration-dependent increase of fetal allergen uptake in all term and preterm placentas. A mean maternal-to-fetal ratio of 20,000/1 and 3,000/1 for BLG, and 40,000/1 and 5,000/1 for OVA was found in term and preterm placentas, respectively. Preterm placentas (27-36 weeks of gestation) were found to favor the diaplacental passage of nutritive allergens compared with placentas at term (>36 weeks of gestation). CONCLUSION: Maternal-to-fetal allergen transport occurs in a dose-dependent and molecular weight-dependent manner with clear accentuation in preterm placentas. PMID- 12576734 TI - Induction of basophil desensitization in physiological medium: enhancement after IgE-dependent upregulation of surface IgE binding on basophils. AB - BACKGROUND: Although the ability of basophils to release mediators, called releasability, may be an important aspect which influences the proinflammatory role of these cells, clinical approaches aiming at the depletion of the releasability have not been established. We examined whether the desensitization procedure in Ca(2+)-containing physiological conditions can make basophils completely unresponsive to IgE-mediated stimulation, and whether basophil desensitization is affected by the surface IgE levels. METHODS: Human peripheral blood basophils were cultured with low concentrations of anti-IgE antibody or recombinant mite allergen. Following culture, cells were stimulated and their histamine release was measured. RESULTS: Culturing with mite allergen or anti-IgE antibody below threshold concentrations induced potent desensitization in basophils. The desensitizing effect of anti-IgE was dose- and time-dependent; IgE dependent releasability was completely suppressed when basophils were incubated with a near-threshold concentration of anti-IgE for > or= 4 h. In the continuous presence of subthreshold doses of anti-IgE, basophils remained desensitized even after 3 days. Basophils which had undergone an increase in surface IgE levels after 24-hour culture with IgE demonstrated enhanced desensitization. CONCLUSIONS: Near-threshold stimulation in physiological medium can affect basophils, thereby inducing complete and sustained deprivation of releasability without triggering degranulation. Basophil desensitization is regulated by their surface IgE levels. Induction of full desensitization may represent a potentially important therapeutic strategy for IgE-mediated allergic diseases in which basophils play pathogenic roles. PMID- 12576735 TI - Comparison of conjunctival and nasal provocation test in allergic rhinitis to house dust mite. AB - BACKGROUND: Nasal allergen provocation tests (NPTs) are useful in confirming the diagnosis of allergic rhinitis, if data obtained by clinical history, skin tests and specific IgE determinations are not conclusive. Since NPTs are laborious, conjunctival provocation tests (CPTs) appear as an attractive alternative. The concordance of CPTs and NPTs with house dust mite allergen extract in sensitized and nonsensitized subjects should be evaluated. METHODS: 50 otherwise healthy subjects with self-reported house dust mite allergy and positive skin prick tests and serum specific IgE to Dermatophagoides pteronyssinus and 45 sex- and age matched healthy controls without allergic symptoms were included. For NPTs, 100 microl allergen extract [10,000 allergy units (AU/ml)] were applied to the less congested nasal cavity. A clinical symptom score and active anterior rhinomanometry were employed to assess the response. For CPTs, 50 microl low concentrated D. pteronyssinus extract (1,000 AU/ml), and if negative, 50 microl normally concentrated extract (10,000 AU/ml) were applied to the lower conjunctival sac. The response was assessed employing clinical symptom scores. RESULTS: NPTs and CPTs yielded concordant results in 90% of the subjects successfully tested (Cohen's kappa = 0.78, p < 0.0001). The diagnostic efficacy of the CPT, with the NPT as the reference method, was 89%, whether or not conjunctival symptoms had been reported in addition to rhinitis symptoms. Both techniques were judged almost equally uncomfortable. CONCLUSION: CPTs are an acceptable alternative to NPTs in patients with allergic rhinitis to house dust mite, even if they have no conjunctival symptoms. PMID- 12576736 TI - Positive effects of a fresh air filtration system on hay fever symptoms. AB - BACKGROUND: Avoidance of allergen exposure is the basic preventive measure in reducing symptoms of immediate-type allergies. As room air cleaners were found to have no benefic effect on general health, their use has not been recommended. The purpose of this study was to evaluate the effect of a fresh air filtration system installed in the bedrooms of non-air-conditioned homes on hay fever symptoms. METHODS: In a placebo-controlled, double-blind, two-period, crossover study, we investigated the effectiveness of a fresh air filtration system installed in the bedrooms of volunteers suffering from hay fever. RESULTS: Volunteers only suffering from pollinosis reported an improvement of their allergy symptoms. We observed a significant decrease in nighttime hay fever symptoms and an increase in morning peak expiratory flow rates (PEFR). No effects were recognized in volunteers who additionally had perennial allergy. CONCLUSION: Based upon these results, the installation of fresh air filtration systems in bedrooms should be recommended for patients suffering from hay fever. PMID- 12576737 TI - No priming of the immune response in newborn brown norway rats dosed with ovalbumin in the mouth. AB - BACKGROUND: Other researchers have reported that the specific immune response to subsequent antigen challenge is primed in newborn mice or rats dosed orally by gavage. We wanted to investigate if priming of a subsequent specific IgE response could be achieved by dosing newborn rats orally with ovalbumin and if this method could be used in an animal model for food allergy. METHODS: Newborn Brown Norway rats were dosed with ovalbumin in the mouth (100 microg or 6 mg). As young adults, the animals were dosed by gavage for 35 days with 1 mg ovalbumin/day or once intraperitoneally with 100 microg. Control groups were dosed by gavage or intraperitoneally but not as neonates. Additionally, young adult rats were dosed with 1 mg ovalbumin/day in the mouth for 35 days. Sera from individual animals were analysed for specific IgE and specific IgG. RESULTS: In all experiments with neonatal rats the specific IgE and IgG responses were decreased compared to the control groups, however, not always reaching statistical significance. A statistical significant decrease in the specific immune response was found in young adult rats dosed in the mouth as compared to by gavage. CONCLUSIONS: Dosing Brown Norway rats with ovalbumin in the mouth as neonates do not prime the specific immune response. The decrease in immune response found in our experiments when dosing newborn animals in the mouth in opposition to the priming seen by others when dosing by intragastric intubation may be explained by a dissimilar antigen presentation when dosing includes both oral mucosa and gut. PMID- 12576738 TI - Immunologic interaction between infiltrating eosinophils and T lymphocytes in murine spontaneous eosinophilic myocarditis. AB - BACKGROUND: Eosinophilic myocarditis often occurs spontaneously in DBA/2 mice. Relationships between infiltrating eosinophils (Eos), T lymphocytes and interleukin-5 (IL-5) in this disorder were investigated microscopically and immunohistochemically. METHODS: Hearts from male DBA/2 mice were studied from 5 to 10 weeks of age. Anti-CD4 and anti-CD8 antibodies were used. Infiltrating Eos, white blood cells (WBC), and CD4+ and CD8+ T cells were counted. Interactions were assessed with multiple regression and forward stepwise regression tests. Additionally, IL-5 distribution in heart tissue was histologically observed with special reference to immunological findings. RESULTS: Beginning at 5 weeks, several necrotic foci containing many damaged cardiocytes were seen in the epicardium of the right ventricle. Eos and lymphocytes were numerous within and around the necrotic foci. Eos were occasionally seen adjacent to degenerating cardiocytes. Three quarters of the mice exhibited such Eo-related myocardial damage. CD4+ lymphocytes were often seen infiltrating actively inflamed foci where Eos could also be observed. The lesional CD4+-to-CD8+ ratio (CD4/CD8) was 5.2 +/- 3.3 at 6 weeks, 6.8 +/- 3.7 at 7 weeks, and 1.5 +/- 0.8 at 10 weeks of age. The lesional Eo/WBC ratio was directly proportional to the CD4/CD8 ratio (p < 0.05), and was also significantly related to lesional CD4+ T cell count (p < 0.05). IL-5 was also expressed in Eo-rich areas. CONCLUSIONS: In this mouse strain with susceptibility to eosinophilic myocarditis, Eo infiltration was related to increased lesional CD4+ cell count. We suspect that CD4+ T cells induce local eosinotaxis, mediated by IL-5, and participate in myocardiocyte injury via Eo induction. PMID- 12576739 TI - Improved mRNA quantitation in LightCycler RT-PCR. AB - BACKGROUND: Real-time polymerase chain reaction (PCR) utilizing the LightCycler and similar systems is an increasingly used technique for quantitative reverse transcription (RT)-PCR of mRNA levels from genes of immunologic interest. A commonly encountered limitation with these systems is that the fluorescence induced by SYBR Green (a fluorophore that binds double-stranded DNA) can result from primer dimers (PDs) as well as the PCR product of interest, thus interfering with the ability to reproducibly quantitate mRNA levels. METHODS: We use a modification of the LightCycler PCR strategy to overcome this problem by altering the PCR strategy to take advantage of the LightCycler's ability to measure fluorescence at a temperature greater than the melting point of PDs. The resulting measurements determine fluorescence of only the desired PCR product. RESULTS: We demonstrate that by using this modified PCR strategy, one can eliminate the fluorescence induced by PDs and obtain accurate product quantitation. CONCLUSIONS: This simple modification allows more precise quantitation of sample mRNA levels by eliminating the contaminating fluorescence induced by the formation of PCR PDs. This modification obviates the need to redesign PCR primers in RT-PCR experiments where this is impractical or impossible. PMID- 12576740 TI - History of fetal diagnosis and therapy: Children's Hospital of Philadelphia experience. AB - Since its inception in 1995, the mission of the Center for Fetal Diagnosis and Treatment at the Children's Hospital of Philadelphia (CHOP) has been to provide comprehensive, multidisciplinary expertise in all facets of prenatal diagnosis, reproductive genetics, and prenatal, perinatal, and postnatal treatment for abnormal fetuses. Most of the authors were trained and/or served on faculty at centers in San Francisco (N.S.A., A.W.F., T.M.C., L.J.H.) or Detroit (A.W.F, M.P.J.). Accurate prenatal detection by serial sonographic study and the development of ultrafast magnetic resonance imaging (MRI) scanners has permitted delineation of the natural history of anatomic malformations. Definition of the pathophysiological features that affect clinical outcome and formulation of management based on prognosis has allowed families to make informed decisions. Outcomes have been improved through carefully planned and coordinated perinatal management. In some cases of life-threatening or severely debilitating defects, prenatal surgery has been offered. PMID- 12576741 TI - Successful treatment of supraventricular tachycardia exhibiting hydrops fetalis with flecainide acetate. A case report. AB - BACKGROUND: The efficacy of flecainide acetate for the treatment of fetal supraventricular tachycardia with hydrops fetalis and changes in venous blood flow patterns in the fetus during treatment are reported. CASE: Oral flecainide administration was started at 30 weeks of gestation. Cardioversion was achieved 6 days after treatment. Sustained abnormal venous Doppler indices were shown and complete normalization of venous returns was observed 6 days after cardioversion. A vigorous male baby was born, and he is now 1 year of age and in good condition with no medication. CONCLUSION: Reversible cardiac dysfunction was observed even after cardioversion in the fetus with supraventricular tachycardia, which could be assessed precisely by venous Doppler analysis. PMID- 12576742 TI - A case of fetal vesicoallantoic cyst in the umbilical cord. AB - A vesicoallantoic cyst in the umbilical cord was detected by fetal ultrasonography at 16 weeks of gestation. The blood flow through the umbilical vessels was evaluated with periodic color Doppler ultrasonographic examinations because compression by the cyst may cause reduction in blood flow and lead to fetal death. After disappearance of the cyst, the infant was delivered vaginally. Retrograde cystography revealed a narrow channel between the bladder and the navel, confirming patent urachus, which was resected without any complications. PMID- 12576743 TI - Feticide during second- and third-trimester termination of pregnancy: opinions of health care professionals. AB - OBJECTIVE: To study the opinions of professionals on feticide being performed as the first step of late termination of pregnancy (TOP). SETTING: Tertiary care obstetrical unit with policy of routine feticide in late TOP. METHOD: Questionnaire survey. RESULTS: 101/109 professionals responded (23 midwives, 22 doctors, 24 nurses, 21 auxiliaries, 9 others). 90 had heard of feticide, 83 knew about how and 38 about when the procedure was done. When asked about what the goals of feticide were, 94 respondents quoted, 'preventing parents from facing neonatal agony', 73 'avoiding fetal pain', 85 'preventing labor ward staff from facing neonatal agony', and 60 'complying with legal rules'. 54 respondents thought feticide was beneficial to their patients or improved their own professional practice, and 71 declared the procedure was emotionally positive, yet stressful. 48 respondents declared lacking information on feticide. Respondents who had attended to feticide at least once (n = 59) knew more on feticide, and were more positive on the impact feticide had on their practice than those who had never attended a feticide (n = 42). CONCLUSIONS: In a highly specialized center, professionals had positive opinions on feticide, expecting it would avoid fetal or neonatal agony and pain. PMID- 12576744 TI - Increased levels of interleukin-6 in cervical secretions and assessment of the uterine cervix by transvaginal ultrasonography predict preterm premature rupture of the membranes. AB - OBJECTIVE: The objective of this study was to investigate the predictive factors of premature rupture of the membranes (preterm PROM). METHODS: The study was undertaken with cervical secretions collected from 72 consenting singleton pregnant women between 20 and 33 weeks of gestation. The levels of interleukin (IL) 1alpha, IL-1beta, IL-6, IL-8, matrix metalloproteinase (MMP) 1, MMP-2, MMP 9, tissue inhibitors of matrix metalloproteinase (TIMP) 1, TIMP-2, granulocyte elastase, and fetal fibronectin in cervical diluted specimens were measured by immunoassay, and the uterine cervix was assessed by transvaginal ultrasonography. Demographic, obstetric, clinical, neonatal, and laboratory data were analyzed by univariate analysis, multiple logistic regression, and receiver operator characteristic curve analysis. RESULTS: Preterm PROM occurred in 6 women, and 63 women delivered at term. Multiple logistic regression analysis indicated a significant independent association with preterm PROM for the cervical IL-6 levels and cervical length. The receiver operator characteristic curve analysis revealed that an IL-6 level of >/=240 pg/ml in cervical secretions and a cervical length of or =0.07 IU/ml showed a higher prevalence of allergic disorders at 3 years of age than those with a cord blood Dp-IgE level of <0.07 IU/ml. These observations suggest that the cord blood Dp-IgE level may be related to allergic manifestations in infancy. PMID- 12576754 TI - Increased respiratory burst and increased expression of complement receptor-3 (CD11b/CD18) and of IL-8 receptor-A in neutrophil granulocytes from newborns after vaginal delivery. AB - To study neutrophil activation in cord blood as a function of the mode of delivery, we performed analysis of the function of neutrophil granulocytes by assessing their ability to produce reactive oxygen products (ROP) as well as neutrophil cell surface expression of CD11b/CD18 and interleukin (IL)-8 receptors quantified with flow cytometry. Plasma levels of granulocyte colony-stimulating factor (G-CSF) were measured using an immunoassay. Neutrophil granulocytes were derived from cord blood of term newborns delivered vaginally (n = 20) and by cesarean section (n = 10), and, for comparison, from adult peripheral blood (n = 15). Blood neutrophil counts and the capacity of neutrophil granulocytes to produce ROP in response to stimulation with Escherichia coli was increased in newborns after vaginal delivery as compared to newborns delivered by cesarean section. The level of expression of the adhesion molecule/complement receptor CD11b/CD18 and the chemokine receptor IL-8 RA was also higher after vaginal delivery. Plasma concentrations of G-CSF in cord blood of newborns were higher than those of adults with no difference detectable between vaginal delivery and cesarean section. The data demonstrate a higher functional responsiveness and a higher expression level of functionally important receptors in neutrophils after vaginal delivery possibly due to activation of neutrophil granulocytes during labor. PMID- 12576755 TI - Endogenous nitric oxide production in the airways of preterm and term infants. AB - Few studies have measured endogenous nitric oxide exhaled from the respiratory system of newborn infants. We measured exhaled nitric oxide (eNO) in the first 48 h of life in 24 (13 preterm, 11 term) spontaneously breathing (online method) newborns using a chemoluminescence analyzer. There was a significant difference in the eNO concentration between term and preterm healthy infants in the first 2 days of life (repeated measures analysis of variance, p < 0.05). In term infants there is a peak eNO production in the first hours of life, suggesting a potential role in postnatal adaptation, while in preterm infants eNO production is almost absent at birth, and then gradually increases. PMID- 12576756 TI - Assessment of the single-occlusion technique for measurements of respiratory mechanics and respiratory drive in healthy term neonates using a commercially available computerized pulmonary function testing system. AB - In this study, a critical assessment of the single-occlusion technique as a means of measuring passive respiratory mechanics and respiratory drive (P0.1) was performed in nonintubated spontaneously breathing healthy term neonates using commercially available computerized equipment (PEDS system). In general, we found that quality parameters only partially conformed to the international established standards for measuring passive respiratory mechanics. There was a failure rate of technically acceptable occlusions of about 50% for respiratory mechanics measurements and 20% for P0.1 measurements. Intersubject variability of the data was between 10 and 40%. After studying different lengths of occlusion times (Tocc; 0.2, 0.3, 0.4 and 0.5 s), it was found that acceptable occlusions for passive respiratory mechanics were obtained only if the expiratory Tocc exceeded 0.2 s. Increasing the Tocc had no effect on the absolute values of respiratory mechanics but showed a significant trend towards more technically acceptable occlusions. We speculate that a Tocc of at least 0.4 s may be a prerequisite for obtaining reliable results for respiratory mechanics in healthy term neonates. PMID- 12576757 TI - Maturation alters cyclic nucleotide and relaxation responses to nitric oxide donors in ovine cerebral arteries. AB - To examine the hypothesis that maturation modulates nitric oxide (NO)-induced relaxation in cerebral arteries, we quantified concentration-relaxation relations and the corresponding dynamic responses of guanosine 3':5'-cyclic monophosphate (cGMP) and adenosine 3':5'-cyclic monophosphate (cAMP) levels following administration of nitroglycerin and S-nitroso-N-acetyl-penicilamine (SNAP), an NO donor, in posterior communicating and middle cerebral arteries from newborn (3-7 days) and adult sheep. The results offer 5 main observations: (1) the efficacy and potency of NO donors were generally greater in newborn than in adult cerebral arteries; (2) rates of relaxation, and presumably rates of NO release, were faster for equimolar concentrations of SNAP than for nitroglycerin in both newborn and adult arteries; (3) basal concentrations were greater for cAMP than for cGMP, and both were greater in newborn than adult cerebral arteries; (4) in adult cerebral arteries, NO-induced increases in cGMP occurred faster but relaxation developed more slowly than in newborn cerebral arteries, and (5) responses to NO donors involved significant cross-reactivity between cGMP and cAMP, the characteristics of which were age, artery, and agent specific. From these results, we conclude that postnatal changes in reactivity to NO reflect corresponding changes in soluble guanylate cyclase activity and possible decreases in NO half-life. We also conclude that maturation slows the mechanisms mediating NO-induced relaxation, and that this effect is more pronounced in distal than in proximal cerebral arteries. The data also suggest that the rate limiting step governing rates of response to NO is probably downstream from cGMP synthesis. From the basal cyclic nucleotide levels, we conclude that basal ratios of synthesis to hydrolysis were greater in fetal than adult arteries. Because NO increased both cGMP and cAMP, we speculate that Type III phosphodiesterase has a possible influence upon cerebrovascular responses to NO, and that this influence varies with postnatal age and artery type. Together, these findings emphasize that the cerebrovascular effects of NO are highly age dependent and artery specific, and should be carefully considered when administering NO therapeutically in the neonate. PMID- 12576758 TI - Influence of changes in dietary fatty acids during pregnancy on placental and fetal fatty acid profile in the rat. AB - To determine whether the composition of long-chain polyunsaturated fatty acids (PUFA) could be modified in the fetus by maternal dietary fatty acids, pregnant Sprague-Dawley rats were fed semipurified diets that differed only in the non vitamin lipid component. The diets contained either 10 g palm, sunflower, olive or fish oil (FOD)/100 g diet. A total of 5-6 rats were studied in each group. At day 20 of gestation, corresponding to 1.5 days prior parturition, the fatty acids in maternal adipose tissue were closely related to the fatty acid composition in the corresponding diet. An important proportion of arachidonic acid (AA) appeared in maternal liver and plasma, although it was lower in the FOD than in the other groups. Except for saturated fatty acids, the proportion of individual fatty acids in the placenta correlated linearly with that in maternal plasma. Also, PUFA in fetal plasma and liver showed significant correlations with PUFA in maternal plasma. Again, AA showed the lowest proportion in the plasma and liver of the FOD group. Therefore, the maternal dietary fatty acid composition influences maternal and fetal plasma and tissue composition, and an increase in dietary omega-3 fatty acids decreases the amount of AA in maternal and fetal tissues. PMID- 12576759 TI - Effect of global hypoxia-ischaemia followed by 24 h of mild hypothermia on organ pathology and biochemistry in a newborn pig survival model. AB - BACKGROUND: Perinatal asphyxia may lead to multiorgan damage as well as brain injury. Posthypoxic hypothermia (HT) may protect other organs in addition to the brain. The aim of this study was to assess the systemic effects of our global hypoxic-ischaemic (HI) insult and compare the effect of mild 24-hour HT with normothermia (NT) during unsedated recovery. METHOD: Thirty-eight newborn pigs were subjected to 45 min of global HI by ventilating them with approximately 6% O2. On reoxygenation, pigs were randomised to NT or HT. The 18 NT piglets were maintained at rectal temperature 39.0 degrees C for 72 h. Twenty-three HT pigs (20 experimental HT and 3 sham controls) were cooled to rectal temperature 35 degrees C for 24 h before NT was resumed and the animals then survived a further 48 h. RESULTS: All lesions were small with no apparent clinical effect. The incidence of any damage to the heart (6 HT vs. 9 NT), liver (9 HT vs. 7 NT), kidney (6 HT vs. 9 NT) or intestinal injury (8 HT vs. 2 NT, p = 0.07) was not different in the two groups. More HT piglets developed lung injury, 10 HT and 3 NT. Plasma [Na], [K], [Ca] and [Mg] increased significantly after the HI insult as compared to baseline values. For the 24-hour period plasma [K] and [Ca] were significantly higher in the HT group, the mean area under the curve (AUC) being for [K] AUC(HT) 4.4 mmol/l vs. AUC(NT) 3.9 mmol/l, p = 0.04 and for [Ca] AUC(HT) 2.7 mmol/l vs. AUC(NT) 2.5 mmol/l, p = 0.01, respectively. Aspartate aminotransferase peaked at 48 h in the HT group and at 24 h in the NT group. Creatinine peaked at >72 h in the HT pigs and at 48 h in the NT pigs. White blood cells (WBC) peaked at 12 h for the HT pigs and at 6 h for the NT animals. AUC of the WBC during the cooling was significantly lower in the HT pig (AUC(HT) 11.1 vs. AUC(NT) 15.3 10(3)/mm3, p = 0.04). The HT pigs needed more glucose to maintain normal glucose during the last 12 h of HT. Also HT animals needed more oxygen during cooling to maintain PaO2. CONCLUSION: Twenty-four hours of mild HT did not reduce damage in any organ. There was a slight increase in lung damage in the HT group. None of the biochemical or pathological changes were of clinical significance. We conclude that mild HT for 24 h does not affect the organ systems adversely when compared to NT. Additional glucose and oxygen is needed during cooling to maintain normal values. PMID- 12576760 TI - [The role of the regional (intra-arterial) chemotherapy in the treatment of hepatic metastases of colorectal cancer]. AB - This focuses on review one of the methods of locoregional treatment - intraarterial hepatic infusion. Metastatic hepatic malignancies are the leading cause of cancer death. Surgical resection of metastatic hepatic malignancies has been the only established treatment modality offering potential for cure. Although surgical resection has significantly improved survival, only 5-20 percent of patients with colorectal carcinoma metastatic to the liver are surgical candidates. Conventional systemic (intravenous) chemotherapy with fluoropirimidines is effective only for 10-21 percent of patients with metastatic colorectal carcinoma. The limitations of surgical resection and the limited efficacy and generalized toxicity of systemic chemotherapy have sparked considerable interest in intraarterial hepatic infusion and especially in combination of systemic and intraarterial chemotherapy. PMID- 12576761 TI - [The importance of left bundle branch block in the diagnosis of acute myocardial infarction]. AB - Electrocardiographic (ECG) evidence of cardiac ischemia or infarction is difficult to detect in the presence of left bundle branch block (LBBB). Traditional ECG indicators of ischemia, such as ST- segment elevation, are common in LBBB and may not indicate acute ischemia. Proper evaluation of the initial ECG is crucial in selecting candidates for early thrombolysis, because the earlier reperfusion treatment is administred, the better are the results. Individuals with LBBB are particularly important stratum of patients to identify. This is true not only because they have a high baseline mortality and receive the greatest incremental improvement in survival when given thrombolytic agents but also because it is tendency to undertreat them. The criteria of Sgarbosa are too insensitive to be used as screening (roule out) test to determine which patients with an LBBB do not have an AMI. The Sgarbosa criteria are, however, highly specific and can be used reliably as confirmatory test to rule in AMI in patients with LBBB. ECG alone doesn't support the diagnosis of AMI. Elevated value of biochemical markers of myocardial necrosis in the presence of LBBB confirms the diagnosis. Despite the recently updated joint practical guidelines of American Heart Association (AHA) and American College of Cardiology (ACC) which defines that all patients having symptoms consistent with acute MI and LBBB should be treated like ST-segment elevation, only minority of them receive thrombolytic therapy, particularly the elderly (only 4%). In the absence of definitive diagnosis of AMI doctors withhold from decision to administer thrombolytic treatment because of risk of haemorrhagic complications. There are not perfect diagnostic tools allowing early diagnostic of AMI in patients having LBBB. Currently the best justified strategy is to follow AHA/ACC recommended guidelines to administer thrombolysis to all patients with LBBB presenting with chest pain, particularly if serum biomarkers are elevated. PMID- 12576762 TI - [Effect of high doses of atorvastatin on the endothelial function of the coronary arteries]. AB - OBJECTIVE: The aim of this study is to investigate the role of high doses of atorvastatin (80 mg per day) in the treatment of endothelium dysfunction of the coronary arteries. MATERIAL AND METHODS: Intracoronary acethylcholine test was performed for 28 (16 in treatment group and 12 in control group) patients who had normal coronary angiogram or minimal lesions (stenosis <50%) and symptomatic coronary heart disease. Acetylcholine was given into left coronary artery in three stages (2 min each) at estimated intracoronary concentrations 10(-6) mol/l, 3.3x10(-6) mol/l,and 10(-5) mol/l. Treatment group patients were given atorvastatin 80 mg per day. Intracoronary acetylcholine test was repeated after six months. The target segment was the segment that exhibited the maximal constrictive response at baseline in a given patient. RESULTS: Total cholesterol was reduced by 40.6+/-11.2% (p<0.001), low density lipoproteins cholesterol by 55.4+/-14% (p<0.001), triglycerides by 29.6% (p<0.05) and high density lipoproteins cholesterol was increased by 24.7% (p<0.05) in atorvastatin group after 6 months of therapy. No significant lipid changes were observed in the control group. Sixteen target segments in the treatment group and 12 segments in the control group were selected for primary efficacy parameter. The vasoconstrictor reaction to acetylcholine was similar at baseline in both groups: -16.8%+/-11.6% of diameter in atorvastatin group and -14.2%+/-13.7% in control group (p=0.58) at dose 10(-6) mol/l, -43.5%+/-21.2% in atorvastatin group and 41.2%+/-22.4% in control group (p=0.79) at dose 3.3x10(-6) mol/l, -64.5%+/-12.4% in atorvastatin group and -57.4%+/-15.6% in control group (p=0.35) at dose 10(-5) mol/l. Treatment with high doses of atorvastatin markedly decreased acetylcholine - induced vasoconstriction: -6.5%+/-14.5% of diameter in atorvastatin group versus -18.3%+/-14.0% in control group (p=0.04) at dose 10(-6) mol/l, -19.7%+/ 22.8% in atorvastatin group versus -40.0%+/-25.0% in control group (p=0.034) at dose 3.3x10(-6) mol/l, -29.9%+/-21.3% in atorvastatin group (6 segments) versus 48.7%+/-19.0% in control group (3 segments, p=0.24) at dose 10(-5) mol/l. CONCLUSION: Treatment with high doses of atorvastatin significantly improves endothelium-mediated response to acetylcholine in the coronary arteries of the patients with minimal lesions. PMID- 12576763 TI - [Drug-resistant tuberculosis in Kaunas region 1997-2001]. AB - Two thousand four hundred thirty nine cases of tuberculosis were confirmed by the method of culture of the sputum at Kaunas Romainiai tuberculosis hospital during the period of 1997-2001. Out of them, 669 of them were diagnozed for the first time (not treated or treated for less than 1 month). Out of confirmed cases, 1601 of confirmed cases were the "old" ones (treatment failures, relapses, treated with one or two courses of antimicrobial therapy). The resistance to H, R, E, S and multidrug resistance have been investigated. We have revealed that among the "new" cases of tuberculosis the resistance of M. tuberculosis has decreased. In 1997 the resistance to H was 27.5 percent. However, it decreased 5.3 percent in 2000 and to 8.7 percent in 2001. In 1997 the resistance to R was 19.9 percent and it has decreased to 9.0 percent and 1.9 percent in 1998 and 1999, respectively. No R resistance cases were diagnozed in 2000 and 2001. In 1997 the resistance to E was 1.1 percent. It reached 1.5 percent in 1998 and there was no resistance to E in 1999-2001. In 1997 the resistance to S was 15.6 percent. In 1998 it was 26.3 percent and dropped to 1.8 percent in 2000. PMID- 12576764 TI - Quantitative analysis of professionally trained versus untrained voices. AB - The aim of this study was to compare healthy trained and untrained voices as well as healthy and dysphonic trained voices in adults using combined voice range profile and aerodynamic tests, to define the normal range limiting values of quantitative voice parameters and to select the most informative quantitative voice parameters for separation between healthy and dysphonic trained voices. Three groups of persons were evaluated. One hundred eighty six healthy volunteers were divided into two groups according to voice training: non-professional speakers group consisted of 106 untrained voices persons (36 males and 70 females) and professional speakers group--of 80 trained voices persons (21 males and 59 females). Clinical group consisted of 103 dysphonic professional speakers (23 males and 80 females) with various voice disorders. Eighteen quantitative voice parameters from combined voice range profile (VRP) test were analyzed: 8 of voice range profile, 8 of speaking voice, overall vocal dysfunction degree and coefficient of sound, and aerodynamic maximum phonation time. Analysis showed that healthy professional speakers demonstrated expanded vocal abilities in comparison to healthy non-professional speakers. Quantitative voice range profile parameters- pitch range, high frequency limit, area of high frequencies and coefficient of sound differed significantly between healthy professional and non professional voices, and were more informative than speaking voice or aerodynamic parameters in showing the voice training. Logistic stepwise regression revealed that VRP area in high frequencies was sufficient to discriminate between healthy and dysphonic professional speakers for male subjects (overall discrimination accuracy--81.8%) and combination of three quantitative parameters (VRP high frequency limit, maximum voice intensity and slope of speaking curve) for female subjects (overall model discrimination accuracy--75.4%). We concluded that quantitative voice assessment with selected parameters might be useful for evaluation of voice education for healthy professional speakers as well as for detection of vocal dysfunction and evaluation of rehabilitation effect in dysphonic professionals. PMID- 12576765 TI - Diagnostic test with omeprazole in patients with posterior laryngitis. AB - The aim of the study was to evaluate the merit of empiric omeprazole therapy in patients with suspected laryngopharyngeal reflux (LPR), to determine the optimal dose and duration of diagnostic test, to compare the diagnostic value of upper gastrointestinal (GI) endoscopy and omeprazole test (OT). One hundred out patients with posterior laryngitis and more than one atypical symptom of LPR, aged 14-68 years were treated with omeprazole for 4 weeks (clinical group). According to received omeprazole dose (20 mg, 40 mg, > 40 mg), three clinical subgroups were selected. Twenty patients treated only with life style modifications and diet composed dietary group. At the entry to the study, a symptom questionnaire (5 laryngeal and 3 esophageal scored from 0 to 3 points), well being in general (W-B(VAS) on 100-mm VAS scale), videolaryngoscopy, upper endoscopy, and voice assessment (4 voice range profile parameters and overall vocal dysfunction degree (VDD)) were completed. Total symptom index (TSI) was calculated multiplying sum of symptoms severity score by number of presented symptoms. Normal values of efficacy parameters were obtained from 113 healthy voice subjects (control group). Patients were evaluated twice during the treatment: after 1-2 weeks and after 4-5 weeks. Patients were confirmed as responders, if TSI improved at least 50%, and patients were satisfied with results. According to our data, the 1st control assessment showed significant improvement on symptoms, laryngoscopy scores, VDD, and W-B(VAS) only for clinical group patients (p<0.05). Responders rate also was advantageous for the clinical group patients in comparison to the dietary group (36.0% vs. 15%). The second control assessment showed significantly better results for the clinical group patients in comparison to the 1st (p<0.05 2nd vs. the 1st). Sixty five percent of them (65/100) were classified as responders (p<0.05). The better results were in patients receiving omeprazole more than 20 mg daily. Erosive esophagitis during upper GI endoscopy was found for 21.0% (21/100) clinical group patients, 18 of them were responders to omeprazole 4-week test (accuracy of OT with regard to confirmed diagnosis with upper endoscopy was 85.7%). At week 4, efficacy parameters were not in normal range. We concluded that short-term treatment with omeprazole might be useful in confirming the clinically based diagnosis of laryngopharyngeal reflux. PMID- 12576766 TI - [Wernicke's encephalopathy induced by hyperemesis gravidarum (case report)]. AB - One of the life-threatening complications of hyperemesis gravidarum is Wernicke's encephalopathy, the main etiological factor of which is a lack of thiamine (vitamin B1). Most frequently Wernicke's encephalopathy is found among persons suffering from excessive drinking. The aim of the report was to present a case of Wernicke's encephalopathy induced by hyperemesis gravidarum. The course of the disease, clinical signs, diagnostics, treatment and its results are presented. Also a review of the literature on Wernicke's encephalopathy secondary to hyperemesis gravidarum is introduced. PMID- 12576767 TI - [Seasonal differences in activity of tRNA and aminoacyl-tRNA synthetases of rabbit liver in myocardial ischemia]. AB - The objective of this study was to examine the acceptor activities of tRNA for amino acids alanine and lysine and activities of corresponding aminoacyl-tRNA synthetases of normal rabbit liver 6, 12 and 24 h after experimental myocardial ischemia in different seasons of the year. MATERIAL AND METHODS: Male rabbits (2.5-3.5 kg) were used. Acute myocardial ischemia was induced by occlusion of the left anterior descending coronary artery. tRNA and aminoacyl-tRNA synthetases were isolated from normal (control) rabbit liver 6, 12 and 24 h after experimental myocardial ischemia in autumn (the months of September and October) and winter (the months of December and January). The acceptor activity of tRNA and activity of alanyl- and lysyl-tRNA synthetases were determined using 14C labelled amino acids alanine and lysine. RESULTS: The results of study show that acceptor activity of rabbit liver tRNA for alanine and lysine under 6, 12 and 24 h experimental myocardial ischemia in autumn is higher by 18-52 percent than in winter. Activities of rabbit liver alanyl- and lysyl-tRNA synthetases under 6, 12 and 24 h experimental myocardial ischemia in autumn is less by 15-35 percent than in winter. No differences in activity of tRNA and aminoacyl-tRNA synthetases between normal groups of both seasons were observed. CONCLUSIONS: The experimental data suggest that there are differences in acceptor activity of rabbit liver tRNA for alanine and lysine and in activity of alanyl- and lysyl tRNA synthetases after 6, 12 and 24 h experimental myocardial ischemia. Decreasing of acceptor activity of tRNA for alanine under experimental myocardial ischemia in winter correlate with increasing of activity of alanyl-tRNA synthetase. Decreasing of acceptor activity of tRNA for lysine under experimental myocardial ischemia correlate with increasing of lysyl-tRNA synthetase activity in both studied seasons. It may be part of the compensatory mechanism of the cell to keep synthesis of protein in normal range under extreme conditions. PMID- 12576768 TI - [Stimulus symmetry influence on curvature perception]. AB - In the psychophysical experiments the symmetrical bright lines of various length and curvature are presented against the dark background on the monitor. The subjects are asked to adjust the test-spot by moving it horizontally or vertically into the invisible arc linking the endpoints of two visible segments of the given stimulus. The data have shown that the precision of curvature estimation depended on type of the curves: it is higher for the centered circle, less for the exponential spiral and still less for the shifted circle. The precision is in direct proportion with the length of the visible segment and in reverse proportion with the invisible segment of the stimulus. The curvature estimation does not depend on the stimulus symmetry, orientation, and position in the visual field. PMID- 12576769 TI - [Development of formulation of hipromellose eye drops (artificial tears)]. AB - The task of this research work was to develop a formulation of hipromellose ophthalmic preparation by determining optimal composition of active ingredients and excipients. Aqueous solution of hipromellose 1% and sorbitol 4.5% was prepared with addition of phosphate buffer (pH 7.05) and preservative cetrimide 0.01%. Sterile filtration was used for solution sterilization. Polimeric membrane prefilters of 1 microm, 0.45 microm and 0.22 microm pore size were employed as sterilizing filter. It is recommended to increase the temperature up to 35-38 degrees C for more efficient filtration of the solution. Quality parameters and analytical methods for quality control have been developed to control the quality and to evaluate the stability of hipromellose eye drops during their storage for 24 months. The obtained results confirmed the sterility and stability of hipromellose eye drops for the period of two years. PMID- 12576770 TI - [Atmosphere air pollution and health of Kaunas children]. AB - Many studies support the impact of long-term exposure to air pollution on respiratory and allergic symptoms and illnesses. The aim of the study was to assess the prevalence of respiratory and allergic symptoms and illnesses in children living in 4 districts of Kaunas, to estimate the indices of respiratory function and to assess the relationship between mentioned indices and air pollutants such as dust, sulfur dioxide and nitrogen dioxide. A cross-sectional survey included 840 Kaunas children (34 subjects in Senamiestis, 288 in Zaliakalnis, 128 in Petrasiunai and 390 in Silainiai) aged 6-7 years, whose parents filled out the questionnaire and children performed respiratory function test. Response rate varied from 58.6 percent to 69.2 percents depending on district. Questionnaire of International Study of Asthma and Allergy in Childhood (ISAAC) was used. The parameters of respiratory function (FVC, FEV1, FEV1/FVC, MEF25, MEF50, MEF75, PEF) were measured with Pony Graphics 3.5. The data showed that prevalence of respiratory and allergic symptoms and illnesses in children living in 4 districts of Kaunas did not differ significantly. In most of the cases it was similar to that found in Riga and Tallinn. There is a significant relationship between MEF25, MEF50, MEF75, PEF and sex as well as gas for cooking. PMID- 12576772 TI - Validate staffing intuitions with a data-rich approach. PMID- 12576771 TI - [Hypothermia]. AB - Hypothermia is defined as a core body temperature less than 35 degrees C (95 degrees F) and results from prolonged exposure to cold environment, drugs, and underlying pathologic conditions. Hypothermia is associated with marked depression of cerebral blood flow and oxygen requirement, reduced cardiac output, and decreased arterial pressure. Victims can appear to be clinically dead because of marked depression of brain and cardiovascular function, bet full resuscitation with intact neurological recovery is possible. Alcohol or drug intoxication are the dominant precipitating factors. There are about 20,000 hypothermia--related deaths a year in Britain, about 25,000--in the USA, 8,000 deaths a year in Canada. There are suggestions that the unofficial number of hypothermia--related deaths is substantially higher, particularity in the elderly. This article reviews the cause, pathogenesis, pathophysiology, clinical features, electrocardiographic manifestations of hypothermia, diagnosis, pre-hospital stabilization, hospital passive, active external, active core rewarming methods, other questions of treatment, and mortality of hypothermia. It is very important to remember, thar if a hypothermic victim is alive when rescued but dies during recovery treatment, and there is no other significant trauma or disease, this suggests that death may have resulted from either inappropriate or ineffective treatment, or no treatment at all. PMID- 12576773 TI - Cardiac markers clarified. PMID- 12576774 TI - Constant change equals constant challenge. AB - Keep your staff up-to-date by posting changes to policies and procedures in your department. PMID- 12576775 TI - IT takes a village to tackle health care's biggest challenges. AB - Examine how initiatives from a host of health care constituencies use information technology to address two of health care's most daunting challenges--the nursing shortage and patient safety. PMID- 12576776 TI - Cautiously delegate medical histories, physical exams. AB - Learn what's expected when delegating medical histories and physical examinations to nurses who aren't licensed independent practitioners. PMID- 12576777 TI - Body adornment: know the limits. AB - Properly developed and implemented dress codes will maintain the professional image of nurses while protecting their legal rights. PMID- 12576778 TI - Called into question: what nurses want. PMID- 12576779 TI - On the trail of community-acquired pneumonia. Recognize the early signs of this respiratory infection to pinpoint effective treatment plan. AB - Learn the significance of prompt identification, treatment, and education of patients with community-acquired pneumonia. PMID- 12576780 TI - The fruits of lifelong learning. Environments rich in continuing education ripen staff development, morale, and retention. AB - Implementation of a mandatory internal continuing education program changed acute care for the better at Wausau Hospital, a 321-bed not-for-profit facility in central Wisconsin. PMID- 12576781 TI - Get real with workload measurement. This innovative technique accurately determines the hours needed for comprehensive patient care. AB - Workload measurement enables clinicians to evaluate work performance and to identify the necessary level of health care resources to efficiently provide patient care. PMID- 12576782 TI - What's the attraction to Magnet hospitals? Unlimited opportunity, resources, and autonomy draw nurses to these empowering facilities. AB - Here, review study results that find nurses employed at Magnet hospitals experience enhanced job satisfaction due to greater access to empowerment structures within their practice setting. PMID- 12576783 TI - What makes the new heart failure guidelines tick? Serving as an adjunct to the NYHA's classification, the ACC/AHA system assesses objective signs of heart disease. AB - A review of the revised ACC/AHA heart failure guidelines. PMID- 12576784 TI - Under pressure: treating cardiac tamponade. Identify this potentially fatal complication early and intervene appropriately. AB - A detailed look at the development and treatment of cardiac tamponade. PMID- 12576785 TI - Computerized order entry goes beyond safety net. AB - Implementing a computerized physician/prescriber order entry at your facility can reduce medication errors and provide staff with clinical decision support tools. PMID- 12576786 TI - Self-care promotes good care. Nurture yourself with the following strategies. PMID- 12576787 TI - A toolbox for healthcare ethics program development. AB - A comprehensive healthcare ethics program in a medical treatment facility develops ethical competence in staff members and creates a climate for ethical practice. This article describes a successful ethics program, provides examples of clinical and organizational ethics issues, outlines activities of an ethics committee, offers a sample outline of a program needs assessment, discusses educational foundations for committee members, and suggests resources to support program development. PMID- 12576788 TI - Building bridges: from high school to healthcare professional. AB - A summer health careers program in existence for 10 years has created interest in high school juniors and seniors to pursue healthcare professions. This program, now open to all students, began as a recruitment initiative to encourage underrepresented minority and disadvantaged students to pursue health careers in a rural North Carolina county, a previous Health Professions Shortage Area (HPSA). Seventy percent (70%) of the 160 students who have participated in the program are pursuing or practicing a health profession, and of these, 50% chose nursing. Collaboration between an Area Health Education Center, a local community hospital, and county school system makes this program successful. The authors describe the history of this program and the importance of collaboration. A framework for the development of a summer program is summarized. Staff development educators can use this framework to develop outreach programs to recruit students into nursing. PMID- 12576789 TI - The phenomenal shift report: a paradox. AB - The shift report, a seemingly routine event, presents a paradox of contradictions for nursing staff. Beyond the acknowledged purpose of information exchange and socialization, intershift reports also include elements of cultural disarray and powerlessness among the nursing staff. This literature review explores the implications for staff development at the unit level and for management. Recommendations are made for enhancing self-esteem and collegial support and for setting priorities for the shift report. PMID- 12576790 TI - Using adaptive competency acquisition assessment for licensed practical nurse career mobility. AB - The purpose of this study was to examine the use of adaptive competency acquisition in Licensed Practical Nurse (LPN) graduate candidates as an assessment mechanism for career mobility. Participants completed the The results indicated that LPN graduate candidates developed adaptive competencies necessary for career transition. The results place value on career mobility, while providing educators insight into the basic competencies of one level of educational preparation, which can aid in service-based program development. PMID- 12576791 TI - Narrowing the possibilities: using quality improvement tools to decrease competence assessment overload. AB - In many institutions, competence assessment tools are seen as the way to address all clinical practice issues. In the absence of a systematic approach to development, these tools are produced en masse with little sharing of information among educators, managers, and clinical leaders. In an effort to harness "competence assessment overload," the authors propose a system that will assist staff in prioritizing the use and development of outcome-driven competence assessment tools. PMID- 12576792 TI - Sharpen customer service skills with PCRAFT Pursuit. AB - Traditional approaches to teaching customer service skills do not involve participant interaction, nor do they provide a fun and relaxed atmosphere for learning. This article describes the development of PCRAFT Pursuit, an innovative game used to teach customer service skills. The development process began with concerns identified through patient satisfaction surveys. The implementation of this game became an integral component of education to improve customer service skills of staff throughout the hospital network. PMID- 12576793 TI - Prognostic factors of renal cell carcinoma. AB - PURPOSE: Determination of prognostic factors is essential for the management of renal cell carcinoma. Stage, histological grade and type, and performance status are now well known and commonly used. During the last decade numerous predictors of patient outcome were tested. This review summarizes the most important studies, explores and compares the results, and tries to respond to the question, "Today, what do we expect of clinical, molecular and genetic factors concerning survival of patients with renal cell carcinoma?" MATERIALS AND METHODS: Based on MEDLINE literature searches we comprehensively reviewed the literature on the prognostic factors associated with the tumor, the patient and the treatment. RESULTS: During the last decades numerous factors have been studied but few of them maintained independent significance in terms of overall survival as assessed by multivariate analysis. Results are more often controversial from one series to another. No known molecular or cytogenetic tumor marker has been identified to help diagnose, manage or confirm renal cell carcinoma remission, progression or relapse. CONCLUSIONS: The classical prognostic factors remain histological grade, histological type, performance status, patient age, number and location(s) of metastatic sites, time to appearance of metastases and prior nephrectomy. The only striking advancement during the last few years has been the proven contribution of radical nephrectomy for metastatic disease in patients with good performance status. PMID- 12576794 TI - The patient's dilemma: prostate cancer treatment choices. AB - PURPOSE: We summarize the literature addressing factors that influence treatment decisions made by men with prostate cancer. MATERIALS AND METHODS: A MEDLINE search of the English language literature published between 1969 and 2000, using the combined MESH key words "prostatic neoplasms," "patient participation," "Internet" and "decision making," generated 181 abstracts. Only 23 of these publications addressed factors influencing treatment decisions made by men with prostate cancer. Nine additional relevant studies were identified from references in the original 23 articles. Subsequently a search for the term "prostate cancer" using several popular Internet search engines yielded more than 1 million hits. A further search was performed using the key words "prostate cancer" and "prostate" within on-line archives of the United Kingdom television channels BBC, ITV, and channels 4 and 5, and newspapers The Sun, The Daily Mail, The Observer, The Guardian and The Times. RESULTS: When there is poor quality evidence or little professional consensus to support a particular treatment over another, no clinical guidelines regarding treatment are possible. Patients are faced with a series of options, and the data reveal that the process of choosing between these options is based on input from a large number of sources. These sources differ in the way that benefits of treatment are emphasized over harms and vice versa. We identified little evidence regarding which type of input exerts the greatest influence on patients. It may be that the sources associated with the most bias have the greatest influence. CONCLUSIONS: There is a paucity of information on how patients with prostate cancer use different types of input in the treatment decision making process. The physician, as principal caregiver, still appears to have the most direct influence on patient choice. Just how long this status will continue is uncertain. PMID- 12576795 TI - Urinary diversion: ileal conduit to neobladder. AB - PURPOSE: The goals of urinary diversion have evolved from simply diverting the urine through a conduit to orthotopic reconstruction, which provides a safe and continent means to store and eliminate urine with efforts to provide an improved quality of life. We address meaningful points that may help optimize clinical results in patients with an orthotopic bladder substitute. MATERIALS AND METHODS: The review involved an objective evaluation of the basic science literature of functional, structural and physiological characteristics of gastrointestinal tissue as a substitute for bladder. Potential problems that may be associated with particular parts of the gut for use in reconstruction are discussed. We also summarize the clinical results and complications of orthotopic reconstruction. RESULTS: In the last 10 years the paradigm for choosing urinary diversion has changed substantially: In 2002 all patients undergoing cystectomy were neobladder candidates. It is critically important to understand the phenomenon of maturation. The motor and pharmacological response of the implanted gut changes dramatically toward that of the bladder. Structural and ultrastructural changes in the ileal mucosa lead to a primitive epithelium similar to urothelium. The need for reflux prevention is not the same as in ureterosigmoidostomy conduit or continent diversion. Reflux prevention in neobladders is even less important than in a normal bladder. When using nonrefluxing techniques, the risk of obstruction is at least twice that after direct anastomosis. Kidney function is not impaired by diversion if stenosis is recognized and managed. Patient health status is more influenced by underlying disease than by diversion. Complications of neobladders are actually similar to or lower than the true rates after conduit formation, in contrast to the popular view that conduits are simple and safe. Some degree of nocturnal leakage is a consistent finding in most reports despite a technically sound operation. The precise pathogenesis of urinary retention requiring clean intermittent catheterization remains uncertain. There are new complications, such as neobladder rupture and mucous tamponade. CONCLUSIONS: Orthotopic reconstruction has passed the test of time. In these patients life is similar to that in individuals with a native lower urinary tract. Until a better solution is devised orthotopic bladder reconstruction remains the best option for patients requiring cystectomy. PMID- 12576796 TI - The natural history of the overactive bladder and detrusor overactivity. A review of the evidence regarding the long-term outcome of the overactive bladder. AB - PURPOSE: The overactive bladder is a widespread medical condition with significant impact on quality of life. We review the existing evidence about the prevalence, impact and long-term followup of the overactive bladder. We also summarize the new International Continence Society terminology associated with the overactive bladder. MATERIALS AND METHODS: A MEDLINE search of all available literature regarding the epidemiology and long-term followup or outcome of the overactive bladder was performed. All articles and relevant resources cited in those articles were reviewed. RESULTS: The new International Continence Society terminology and definitions will allow greater consistency in future research. Two large, recently published surveys of the prevalence of overactive bladder give new insight into the widespread nature of the problem and its significant impact on quality of life. There is little good quality evidence currently available on the long-term outcome of overactive bladder. CONCLUSIONS: There is little published evidence regarding the natural history of overactive bladder. While recent studies have improved understanding the prevalence of the condition, we still know little about long-term outcome. The few published studies suggest that it is a chronic condition that persists urodynamically and symptomatically. PMID- 12576797 TI - Disease progression and survival of patients with positive lymph nodes after radical prostatectomy. Is there a chance of cure? AB - PURPOSE: In prostate cancer involvement of regional lymph nodes is regarded as a poor prognostic factor. Is this also true for micrometastasis if a meticulous lymph node dissection is performed? We determined progression rate and survival of patients with positive nodes following radical prostatectomy according to the number of metastases. MATERIALS AND METHODS: Between 1989 and 1999, 367 patients with clinically organ confined prostate cancer underwent meticulous pelvic lymph node dissection and radical prostatectomy. None of the patients received immediate adjuvant therapy. RESULTS: Of the patients 92 (25%) had histologically proven lymph node metastases. Followup of more than 1 year was available in 88 patients (96%), and median followup was 45 months (range 13 to 141). Of 19 patients (22%) who died of prostate cancer 16 had more than 1 positive node. Of the 39 patients with only 1 positive node 15 (39%) remained without signs of clinical or chemical progression. Whereas of the 20 and 29 patients with 2 or more positive lymph nodes only 2 (10%) and 4 (14%), respectively, remained disease-free. Time to prostate specific antigen relapse, symptomatic progression and tumor related death were significantly affected by the number of positive nodes. CONCLUSIONS: Meticulous lymph node dissection reveals a high rate of metastases (25%). In patients with positive nodes time to progression is significantly correlated with the number of diseased nodes. Some patients with minimal metastatic disease remain free of prostate specific antigen relapse for more than 10 years after prostatectomy without any adjuvant treatment. Meticulous pelvic lymph node dissection, particularly in patients with micrometastases, seems not only to be a staging procedure, but may also have a positive impact on disease progression and long-term disease-free survival. PMID- 12576798 TI - Intermediate outcomes of dual renal allografts: the University of Washington experience. AB - PURPOSE: The increased survival advantage of renal transplantation with end stage renal disease combined with an increasing incidence of renal disease fuel an increasing disparity between supply and demand for transplantable kidneys. Despite efforts to increase cadaveric organ donation through education and publicity, the number of cadaveric kidneys transplanted has not increased and in the last year was surpassed by kidneys transplanted from living donors. In an effort to maximize cadaver organ donors use of kidneys from expanded criteria donors has been investigated. In select cases both donor kidneys have been transplanted into a single recipient, which is called dual renal transplant. We report on the 4-year dual renal transplant graft and patient outcomes and compare these to age matched single cadaver kidney transplants. MATERIALS AND METHODS: A retrospective review of 10 dual renal transplant recipients and 10 age matched single cadaver kidney recipients was performed. All patients underwent transplantation at our university between January 1996 and February 1998. Mean followup was 4.1 years (range 2.5 to 5.1) for the dual kidney recipients and 3.6 (0.0 to 5.5) years for the control group. RESULTS: Of the 10 dual renal transplant recipients 7 remain alive and 3 died of nontransplant related causes. Of the 10 single recipients 8 are alive, 1 died of postoperative complications and 1 died of nontransplant related causes. When censored for death with a functioning graft, 7 of 10 dual grafts are functioning at followup with a mean creatinine clearance of 39.4 ml. per minute (range 16.1 to 65.9) and mean serum creatinine of 2.0 mg./dl. (1.1 to 3.9). If not censored for death with a functioning graft, 50% of dual grafts are functioning. Of the 3 graft losses 2 were due to recurrent disease and 1 was attributed to chronic rejection. In the control group 8 of 10 grafts are functioning at current followup (regardless of censoring for death with a functioning graft) with a mean creatinine clearance of 48.7 ml. per minute (range 23.4 to 66.5) and mean serum creatinine of 1.6 mg./dl. (1.2 to 2.4). Of the 2 graft losses 1 resulted from postoperative complications and 1 was due to chronic rejection. CONCLUSIONS At the 4-year followup patients undergoing dual renal transplant have comparable graft function, incidence of graft loss and survival compared to the control group. However, because of our small sample size, differences in the 2 groups may be significant in a larger study. Additional studies need to be conducted to determine if this practice represents an acceptable use of kidneys from expanded criteria donors. PMID- 12576799 TI - Vascular complications after live donor renal transplantation: study of risk factors and effects on graft and patient survival. AB - We evaluated the incidence and management of vascular complications after live donor renal transplantation. Possible risk factors and their effects on patient and graft survival were also assessed. MATERIALS AND METHODS: A total of 1,200 consecutive live donor renal transplants were performed in 1,152 patients at a single institution. The incidence of different types of vascular complications were determined and correlated with relevant risk factors. The impact on patient and graft survival was also studied. RESULTS: There were 34 vascular complications (2.8%). Stenotic or thrombotic complications were recorded in 11 cases (0.9%), including renal artery stenosis in 5 (0.4%), renal artery thrombosis in 5 (0.4%) and renal vein thrombosis in 1 (0.1%). Hemorrhagic complications were observed in 23 patients (1.9%). Although no risk factors could be identified that were related to stenotic or thrombotic complications, grafts with multiple renal arteries were significantly associated with hemorrhagic complications (p = 0.04). Stenotic and thrombotic complications as well as hemorrhagic complications were significantly associated with subsequent biopsy proved acute tubular necrosis (p <0.001). The mean 5-year patient and graft survival rates +/- SD for those with vascular complications were 71.9% +/- 1.9% and 41.6% +/- 8.9% compared with 86.3% +/- 1.1% and 76.8% +/- 1.4% for the remainder of our transplant population, respectively (p <0.001). The deleterious impact on survival was not only observed in recipients with thrombotic or stenotic crises, but also in those with hemorrhagic sequelae. CONCLUSIONS: Hemorrhagic crises are as serious as the stenotic and thrombotic complications affecting patient and graft survival. Because they are a significant factor in the development of hemorrhagic complications, grafts with multiple renal arteries should be managed critically. PMID- 12576800 TI - Changes in urine volume accomplished by physicians treating nephrolithiasis. AB - PURPOSE: We quantified the changes in urine volume and sodium accomplished in various practice settings and the consequent effects on calcium oxalate supersaturation. MATERIALS AND METHODS: We determined comprehensive urine stone risk factors in 2,877 patients treated in 14 practices, including a university referral center and private sector practices. Changes in urine volume and stone risk factors were measured. RESULTS: In a wide range of practice settings the volume increase was about 0.3 l. daily. Urine sodium excretion increased with volume for unexplained reasons, as did urine calcium excretion. As expected, thiazide lowered calcium excretion but the effect progressively decreased as urine volume increased. Therefore, urine calcium and calcium oxalate supersaturation changes were the result of opposing forces. The net effect was a decrease partly offset by sodium and calcium excretion increases. CONCLUSIONS: Urine volume increments are modest in practice and they are modestly offset by increases in urine sodium due to increased sodium intake. Clinicians should strive for higher volume increases than are currently achieved and be vigilant concerning what seems to be a strong tendency toward a higher sodium intake with more fluids. PMID- 12576801 TI - Influence of urine pH and citrate concentration on the upper limit of metastability for calcium phosphate. AB - PURPOSE: We determined the effects on the urine upper limit of metastability for calcium phosphate of citrate concentration and pH, and achievement of the upper limit of metastability by adding calcium or phosphate. MATERIALS AND METHODS: The citrate concentration in aliquots of 24-hour urine samples from normal males without a history of kidney stones was increased. The upper limit of metastability was determined by the point of visible crystal formation, as confirmed by increased optical density at 620 nm. when calcium or pH was increased. In additional experiments the upper limit of metastability was determined by adding calcium or phosphate at pH 5.9 and 6.4. RESULTS: Regardless of how the upper limit of metastability was achieved increasing the citrate concentration increased the former value by about 0.4 units per mM. citrate per l. The upper limit of metastability achieved in a given urine sample by adding phosphate or calcium did not differ. Increasing urine pH increased the upper limit of metastability. CONCLUSIONS: Treatment with alkaline citrate salts may decrease stone formation via an increase in calcium phosphate upper limit of metastability by increasing urine citrate and by directly affecting increased pH. PMID- 12576802 TI - Variation in urine composition in the human urinary tract: evidence of urothelial function in situ? AB - PURPOSE: An increased awareness of the concept that the urothelium has a significant transport function led us to question whether urine composition changes as it passes along the human lower urinary tract. MATERIALS AND METHODS: Urine samples from the bladder and renal pelvis were collected from 30 adults who underwent percutaneous nephrolithotomy (27) or ureteral stent insertion before lithotripsy (3). Urine was obtained from the 2 renal pelves (operative and contralateral sides) in 6 patients (24%). Urine pH was measured using an ultra thin glass pH electrode. Urinary osmolality, Na and K were measured by micro osmometry and flame photometry, respectively. Comparison of data sets was achieved using conventional nonparametric statistical methods. RESULTS: Median bladder urine pH in 30 patients, osmolality in 16, Na in 16 and K in 15 were significantly higher than in the renal pelvis at 6.76 (IQR 6.23 to 6.99), 469 mOsm. kg.1 (IQR 349 to 553), 132 (IQR 100 to 154) and 45 mM. (IQR 30 to 64) versus 6.08 (IQR 5.84 to 6.89), 308 mOsm. kg.1 (IQR 248 to 465), 90 (IQR 69 to 115) and 17 mM. (IQR 10 to 47), respectively (p < or = 0.05). There was no significant difference in these parameters in the urine of the paired renal pelves. CONCLUSIONS: Bladder urine pH, osmolality, Na and K significantly differ from values in the renal pelvis in moderately hydrated humans. Our data show that urine composition is modified in the lower urinary tract, supporting the concept of a dynamic urothelium. We propose that urothelial-urinary interactions and urinalysis need reappraisal, particularly in investigations of urinary stone formation and sensory bladder function. PMID- 12576804 TI - Extracorporeal shock wave lithotripsy for distal ureteral calculi: what a powerful machine can achieve. AB - PURPOSE: We assessed the efficacy of extracorporeal shock wave lithotripsy (ESWL, Dornier Medical Systems, Inc., Marietta, Georgia) for distal ureteral calculi with the HM3 (Dornier Medical Systems, Inc.) lithotriptor. MATERIALS AND METHODS: A total of 585 consecutive patients with distal ureteral calculi were treated with ESWL using an unmodified HM3 lithotriptor. Of these patients 67 referred for treatment only for whom no followup was available were excluded from further analysis. The remaining 518 cases were followed until they were radiologically documented to be stone-free or considered treatment failures. Before ESWL additional procedures were performed in 144 patients, including stone push back, ureteral catheter or Double-J (Medical Engineering Corp., New York, New York) stent placement, percutaneous nephrostomy, ureteral endoscopic maneuvers or stone basket manipulation. A total of 374 patients needed no preliminary treatment before ESWL. RESULTS: Of the 518 patients 469 (91%) were successfully treated with 1 ESWL session, while 49 (9%) needed 2 or 3. Manipulation after ESWL was performed in 22 cases, including stent placement, percutaneous nephrostomy, ureteral endoscopic stone removal and a stone basket procedure. On day 1 after ESWL 327 patients (63%) were stone-free, 158 (30%) had less than 5 mm. fragments and 33 (7%) had more than 5 mm. fragments. At 3 months the stone-free rate increased to 97%. CONCLUSIONS: These data show that ESWL for distal ureteral calculi with the powerful unmodified HM3 lithotriptor has a high success rate with a low rate of minimally traumatic manipulations before and after intervention. Results in terms of the re-treatment and stone-free rates are superior to those of any other second or third generation lithotriptor and comparable to the results of the best ureteroscopic series. PMID- 12576803 TI - The role of mannitol in alleviating renal injury during extracorporeal shock wave lithotripsy. AB - PURPOSE: We determined the role of mannitol in preventing or alleviating renal injury during extracorporeal shock wave lithotripsy (ESWL, Dornier Medical Systems, Inc., Marietta, Georgia). MATERIALS AND METHODS: Patients undergoing ESWL were randomized to receive mannitol or control. Change in the levels of urinary enzymes, beta 2-microglobulin and microalbumin were compared in the groups before and after the procedure. RESULTS: Mannitol treated patients had a statistically significant decrease in beta 2-microglobulin excretion after ESWL compared with the control group. CONCLUSIONS: Mannitol may serve a protective function by decreasing the amount of renal injury caused by ESWL for renal calculous disease. PMID- 12576805 TI - Stone retropulsion during holmium:YAG lithotripsy. AB - PURPOSE: We modeled retropulsion during holmium:YAG lithotripsy on the conservation of momentum, whereby the force of ejected fragment debris off of the calculous surface should equal the force of retropulsion displacing the stone. We tested the hypothesis that retropulsion occurs as a result of ejected stone debris. MATERIALS AND METHODS: Uniform calculous phantoms were irradiated with holmium:YAG energy in air and in water. Optical fiber diameter and pulse energy were varied. Motion of the phantom was monitored with high speed video imaging. Laser induced crater volume and geometry were characterized by optical coherence tomography. To determine the direction of plume laser burn paper was irradiated at various incident angles. RESULTS: Retropulsion was greater for phantoms irradiated in air versus water. Retropulsion increased as fiber diameter increased and as pulse energy increased (p <0.001). Crater volumes increased as pulse energy increased (p <0.05) and generally increased as fiber diameter increased. Crater geometry was wide and shallow for larger fibers, and narrow and deeper for smaller fibers. The ejected plume propagated in the direction normal to the burn paper surface regardless of the laser incident angle. CONCLUSIONS: Retropulsion increases as pulse energy and optical fiber diameter increase. Vector analysis of the ejected plume and crater geometry explains increased retropulsion using larger optical fibers. Holmium:YAG lithotripsy should be performed with small optical fibers to limit retropulsion. PMID- 12576806 TI - Is a 2-week duration sufficient for stenting in endopyelotomy? AB - PURPOSE: Internal stenting is an integral part of endopyelotomy. Studies in animals show good healing after 1 to 2 weeks of ureterotomy. Inherent stent related problems warrant a minimum possible duration of stenting without compromising the results of endopyelotomy. We performed a prospective randomized trial to evaluate the optimum duration of stenting after endopyelotomy. MATERIALS AND METHODS: A total of 57 consecutive patients with primary ureteropelvic junction obstruction were randomized to undergo 7/14Fr internal endopyelotomy stent placement for 2 (group 1) and 4 (group 2) weeks. A symptom based questionnaire was administered to all patients at stent removal. Followup was done with diuretic scanning at 3, 6, 9 and 12 months and then yearly, and thereafter with diuretic renography. RESULTS: In each group 26 patients were available for evaluation. The 2 groups were comparable in terms of age, sex, symptoms and ipsilateral glomerular filtration rate. Mean followup was 22.3 (range 12 to 36) and 21.3 months (range 12 to 35) in groups 1 and 2, respectively. At the end of 1 year 24 group 1 (92.3%) and 23 group 2 (90.3%) patients had an improved drainage pattern. This difference was not significant. Stent related symptoms were present in a good proportion of patients in groups 1 and 2 but there was a significant difference in the incidence of urinary tract infections (11.5% versus 38.1%, p = 0.04). Of the group 2 patients 64% preferred 2 weeks of stenting. CONCLUSIONS: Two weeks seems to be a sufficient duration to allow functional restoration across the ureteropelvic junction after endopyelotomy and decrease stent related complications. PMID- 12576807 TI - Hand assisted retroperitoneoscopic nephroureterectomy: comparison with the open procedure. AB - PURPOSE: We report our initial experience with hand assisted retroperitoneoscopic nephroureterectomy for upper urinary tract transitional cell carcinoma and compared our results to those of a contemporary series of open nephroureterectomy. MATERIALS AND METHODS: Clinical data on 34 consecutive patients who underwent hand assisted retroperitoneoscopic nephroureterectomy were reviewed and compared with those on 34 who underwent open nephroureterectomy. All specimens were extracted intact. RESULTS: Total operative time was similar in the 2 groups (233 versus 236 minutes). Estimated blood loss was significantly less in the hand assisted laparoscopy group (236 versus 427 ml.). During convalescence the frequency of parenteral analgesia postoperatively was significantly lower in the laparoscopy group than in the open surgery group. The former patients had a significantly shorter interval to oral intake and ambulation as well as a shorter hospital stay and convalescence compared with those who underwent open surgery. In each group the complication rate was 12% (4 of 34 cases). In the hand assisted laparoscopy group 1 conversion to an open procedure was due to bleeding. There was recurrence in 4 of the 34 patients (12%) in the laparoscopy group, including 2 with bladder recurrence and 2 with metastases, at an average followup of 13.1 months. CONCLUSIONS: Hand assisted retroperitoneoscopic nephroureterectomy is an effective and safe alternative to open nephroureterectomy for upper urinary tract transitional cell carcinoma. PMID- 12576808 TI - Surgical management of multi-organ visceral tumors in patients with von Hippel Lindau disease: a single stage approach. AB - PURPOSE: We assessed surgical feasibility of a 1-stage multi-organ approach for multiple visceral tumors in patients with von Hippel-Lindau disease. MATERIALS AND METHODS: A total of 14 men and 15 women with von Hippel-Lindau disease underwent simultaneous multi-organ surgery for multiple adrenal, renal and pancreatic tumors at the National Cancer Institute between 1988 and 2001. Perioperative and followup data were analyzed retrospectively. The Mann-Whitney U test was used for statistical analysis. RESULTS: Surgery involving 2 or more organs (mean 2.4 procedures per patient, range 2 to 4) was performed in all patients and concurrent pancreatic operations were performed in 12 (41%). Overall a combined 71 procedures, were performed including 4 cases (13%) treated laparoscopically. Mean +/- SD operative time and estimated blood loss were 464 +/ 142 minutes (range 206 to 830) and 2,798 +/- 4,285 cc (300 to 20,000), respectively. In 16 patients (55%) blood transfusion was administered intraoperatively. At a median followup of 21 months (range 5 to 151) renal tumors recurred in 8 patients (28%), requiring further kidney operations, but no patient had pancreatic or adrenal recurrence. The overall complication rate was 38%, and there was no operative mortality. CONCLUSIONS: A single stage surgical approach for multi-organ visceral tumors is a viable option for patients with von Hippel Lindau disease. With careful patient selection and surgical planning combined procedures can be safely performed in 1 operative setting. PMID- 12576809 TI - TNM T3a renal cell carcinoma: adrenal gland involvement is not the same as renal fat invasion. AB - PURPOSE: Upper pole tumors with direct extension into the adrenal gland are currently staged as pT3a tumors in the 1997 TNM staging system. To determine whether the clinical behavior of pT3a adrenal tumors differs from that of tumors with perinephric fat invasion (also stage pT3a) a retrospective analysis was performed. MATERIALS AND METHODS: Of 1,087 patients who underwent nephrectomy 27 were identified with direct adrenal involvement and 187 were identified with perinephric fat or renal sinus involvement. Variables and outcomes analyzed in each group included the percent of patients with metastatic disease at presentation, lymph node involvement, Eastern Cooperative Oncology Group score, response to immunotherapy, and median and overall survival using Kaplan-Meier curves. RESULTS: Median survival for patients with pT3a disease and perinephric or renal sinus fat involvement was 36 months with a 36% 5-year cancer specific survival rate. In contrast, patients with adrenal gland invasion had significantly worse survival at a median of 12.5 months and a 0% 5-year cancer specific survival rate (p <0.001), which was similar to median survival of those with stage pT4 disease (11 months). CONCLUSIONS: Upper pole tumors with direct extension into the adrenal gland predict significantly worse survival than similarly staged tumors with fat invasion and they have a prognosis similar to that of stage pT4 disease. While these data await external validation, consideration should be given to re-categorizing tumors with direct adrenal gland involvement as stage pT4 or in a subcategory such as pT4a. PMID- 12576810 TI - Renal cell carcinoma: histological findings on surgical margins after nephron sparing surgery. AB - PURPOSE: We evaluated the incidence of peritumoral satellite lesions in nephron sparing surgery and examined whether these findings have a negative effect on cancer specific survival and on the percent of local recurrence. MATERIALS AND METHODS: We performed nephron sparing surgery in 63 patients with kidney cancer, including 53 elective (group 1) and 10 imperative (group 2) operations. In all cases we removed 10 mm. of apparently healthy peritumoral parenchyma with the tumor. This tissue was subsequently examined by an anatomical pathologist to identify any satellite lesions. RESULTS: Four satellite lesions were identified, including 3 in group 1 and 1 in group 2, at a mean of 5.3 mm. from the primary lesion. None of the patients in either group had local recurrence at followup. Cancer specific survival was 96.3% in group 1 (mean followup 61 months) and 58% in group 2 (mean followup 39 months). It was not influenced by the presence of satellite micro-lesions. CONCLUSIONS: Despite common perplexities concerning the risk of multifocality in renal cell carcinoma we believe that the nephron sparing procedure in select patients is as effective as radical surgery. Based on our experience the surgical safety margin must be at least 10 mm. of macroscopically healthy, peritumoral tissue. PMID- 12576811 TI - Renal cell carcinoma with tumor thrombus extension: biology, role of nephrectomy and response to immunotherapy. AB - PURPOSE: We outline the biology, prognosis and role of immunotherapy for renal cell carcinoma with gross venous tumor thrombus. MATERIALS AND METHODS: A total of 207 patients with unilateral renal cell carcinoma and tumor thrombus into the renal vein (107) and inferior vena cava (100) who underwent nephrectomy and thrombectomy were compared with 607 without tumor thrombus. RESULTS: At diagnosis 77 patients (37%) had N0M0 disease and 130 (63%) had lymph node (N+) or distant (M1) metastases. Compared with nontumor thrombus cases tumor thrombus was associated with more advanced stage, N+ (26% versus 12%), M1 (54% versus 31%) disease, higher grade and Eastern Cooperative Oncology Group performance status. In N0M0 cases with inferior vena caval tumor thrombus capsular penetration, collecting system invasion and extension into the hepatic vein were more important prognostic variables then the level of inferior vena caval thrombus. In patients with confined N0M0 tumors mean 2 and 5-year survival +/- SD was 83% +/- 8.8% and 72% +/- 10.7% in those with inferior vena caval tumor thrombus, and 90% +/- 9.4% and 68% +/- 16.1% in those with renal vein tumor thrombus, similar to the 93.4% +/- 1.7% and 81 +/- 3.1% rates, respectively, in those without thrombus who had no recurrence within 6 months after nephrectomy. Of patients with M1 disease in whom cytoreductive surgery was done those with and without thrombus showed a similar response to immunotherapy. When there was inferior vena caval and renal vein thrombus, mean 2-year survival was higher after nephrectomy and immunotherapy than after nephrectomy alone (41% +/- 9% and 52% +/- 7% versus 32% +/- 13% and 45% +/- 7%), immunotherapy alone (0% and 13% +/- 12%, respectively) and no treatment (0%). CONCLUSIONS: Renal cell carcinoma with tumor thrombus is associated with worse characteristics. Local tumor extension has greater prognostic importance than the level of inferior vena caval tumor thrombus. Survival is fair in patients with truly confined N0M0 disease and thrombus. The combination of surgery and immunotherapy has a role in thrombus cases. Our data provide the rationale for a prospective study of adjuvant immunotherapy after surgery in N0M0 cases with extensive tumor thrombus. PMID- 12576812 TI - Neural network using combined urine nuclear matrix protein-22, monocyte chemoattractant protein-1 and urinary intercellular adhesion molecule-1 to detect bladder cancer. AB - PURPOSE: We developed a neural network to identify patients with bladder cancer more effectively than hematuria and cytology. The algorithm is based on combined urine levels of nuclear matrix protein-22, monocyte chemoattractant protein-1 and urinary intercellular adhesion molecule-1. MATERIALS AND METHODS: A randomized double-blinded study of voided urine from 253 patients undergoing outpatient cystoscopy was performed. Of the patients 27 had bladder cancer on biopsy and 5 had muscle invasion. Urine tumor markers were measured using sandwich-enzyme linked immunosorbent assay kits. Urine from patients with bladder cancer on cystoscopy was compared to urine from controls with negative cystoscopy results. An algorithm was created with 3 sets of cutoff values modeled to be 100% sensitive for superficial bladder cancer, 100% specific for superficial cancer and 100% specific for muscle invasive cancer, respectively. We compared our model to hematuria and cytology. RESULTS: For the hematuria dipstick test sensitivity, specificity, positive and negative predictive values were 92.6%, 51.8%, 18.7% and 98.2%, respectively. For atypical cytology sensitivity, specificity, positive and negative predictive values were 66.7%, 81%, 29.5% and 95.3%, respectively. For the sensitive model set sensitivity, specificity, positive and negative predictive values were 100%, 75.7%, 32.9% and 100%, respectively. For the specific model set sensitivity, specificity, positive and negative predictive values were 22.2%, 100%, 100% and 91.5%, respectively. For the muscle invasive model set sensitivity, specificity, positive and negative predictive values were 80%, 100%, 100% and 99.6%, respectively. The standard bladder tumor evaluation of 253 patients costs 61,054 US dollars but 36,450 US dollars using our model. CONCLUSIONS: Our algorithm is superior to conventional screening tests for bladder cancer. The model identifies patients who require cystoscopy, those with bladder cancer and those with muscle invasive disease. It provides possible savings over current screening methods. The potential loss of other information by not performing cystoscopy was not evaluated in our study. PMID- 12576813 TI - Immunocyt test improves the diagnostic accuracy of urinary cytology: results of a French multicenter study. AB - PURPOSE: The limitations of urinary cytology and the invasiveness of cystoscopy generate an increasing interest in noninvasive diagnostic tools for the management of transitional cell carcinoma. We assess the clinical performance of ImmunoCyt (DiagnoCure, Inc., Saint-Foy, Canada) in the detection of bladder cancer in a 10-center French trial. MATERIALS AND METHODS: From October 2000 to April 2001, 694 patients undergoing cystoscopy were prospectively included in the study. Of the patients 458 (66%) had been previously treated for superficial transitional cell carcinoma and 236 (34%) were referred for symptoms suggestive of bladder cancer. All patients underwent ImmunoCyt test and standard urinary cytology from voided urine as well as a complete evaluation including cystoscopy and transurethral resection or biopsy of suspicious lesions. Sensitivity and specificity values of urinary cytology and ImmunoCyt whether or not combined were calculated using cystoscopy as the gold standard and histopathology when available. RESULTS: A total of 85 recurrent and 58 newly diagnosed bladder tumors were diagnosed by cystoscopy and histologicaly confirmed. Overall sensitivity of urinary cytology was 17.9%, 46.3% and 63.8% respectively, for G1, G2 and G3 transitional cell carcinoma, whereas that of ImmunoCyt was 60.7%, 75.6% and 76.8%. Sensitivity of the combined tests was 66.7%, 78% and 87%, respectively. Moreover, 10 of 55 (18.2%) new pT1 and pT2 or greater tumors were diagnosed by ImmunoCyt alone. Specificity of urinary cytology was 94.5%, whereas that of ImmunoCyt was 84.2%. Specificity of the combined tests was 80.7%. Marked variations in urinary cytology sensitivity were observed among the different centers (27.3% to 66.7%), whereas combined assays (urinary cytology and ImmunoCyt) enhanced the overall sensitivity in the 80% range at most centers. CONCLUSIONS: This prospective multicenter series confirmed a marked increase in sensitivity without significant loss in specificity when including ImmunoCyt in standard urinary cytology protocol. This increased sensitivity was observed in high grade lesions (with 100% sensitivity for carcinoma in situ) as well in low grade, low stage tumors. PMID- 12576814 TI - Percutaneous management of renal pelvic urothelial tumors: long-term followup. AB - PURPOSE: We present the long-term outcome of percutaneous resection of renal urothelial tumor. MATERIALS AND METHODS: A total of 24 patients underwent primary percutaneous resection of renal urothelial tumor. Patients with low stage pT0-1 disease were treated primarily with percutaneous surgery. All pelvicaliceal tumors were taken for biopsy and treated with percutaneous resection. Patients with multi-segmental pelvicaliceal system involvement, stage greater than pT1, high grade histology or additional ureteral tumors were considered for nephroureterectomy. Topical chemotherapy (mitomycin C or epirubicin) was administered via nephrostomy tube or intravesical instillation after Double-J stent (Medical Engineering Corp., New York, New York) insertion. Surveillance included upper tract cytology, nephroscopy or fiberoptic ureterorenoscopy. Long term followup was correlated with histopathology. RESULTS: Of the 24 cases 2 had squamous cell carcinoma, 5 had grade III transitional cell carcinoma, 15 had grade I to II transitional cell carcinoma and 2 had no tumor. Control was established with initial percutaneous resection in 18 (75%) cases and second look nephroscopy in 4. Early recurrences were detected by excretory urography (IVP) in 3 cases, small pelvic recurrences by IVP in 2, fiberoptic ureterorenoscopy in 2 and bladder tumors by flexible cystoscopy in 3 after 1 year. A total of 10 nephroscopies were performed in 5 cases, 24 flexible uretereorenoscopies in 9 and IVP in 6. Three synchronous, grade I bladder tumors were managed conventionally. All patients with high grade disease died of malignancy except one (with no further treatment) and 6 of the 15 patients with low grade noninvasive transitional cell carcinoma underwent nephroureterectomy during followup either due to progression of disease, concomitant tumor or complications. Two patients with solitary kidneys died of renal failure unrelated to malignancy. High grade tumors or tumors greater than T1 were treated with nephroureterectomy early during management. There was no perioperative mortality and 9 (60%) of the low grade cases the kidneys were preserved at a mean followup +/- SD of 64 +/- 15 months. All excised tracks from patients who underwent nephroureterectomy and the renal fossae were free of tumor on histopathological examination. CONCLUSIONS: Percutaneous resection of transitional cell tumor should be considered primarily in patients with early stage disease excluding tumors crossing caliceal infundibula, ureteropelvic junction tumor, tumor extending over multiple calices and synchronous ureteral tumors. The long-term outcome of low grade tumors is good and they should be managed by either form of minimally invasive surgery. Nephron sparing is possible in a large percentage of low grade disease but high grade tumors should be treated with nephroureterectomy. PMID- 12576815 TI - T2a transitional cell carcinoma of the bladder: long-term experience with intravesical immunoprophylaxis with bacillus Calmette-Guerin. AB - PURPOSE: In this prospective study we evaluate the effect of combined transurethral resection of early muscle invasive bladder cancer and immunotherapy with bacillus Calmette-Guerin (BCG) in patients unfit for radical cystectomy or refusing more aggressive therapies. MATERIALS AND METHODS: A total of 22 patients with a mean age 73.6 years were included in the study. Inclusion criteria were histologically proven muscle invasive transitional cell carcinoma of the bladder with a tumor-free second resection and negative staging examinations in patients unfit for radical cystectomy or refusing more aggressive therapies. All patients received 6 weekly instillations of 120 mg. BCG starting 14 to 21 days after the last transurethral resection of the tumor. Followup at 3 months included cystoscopy, urinary cytology, ultrasound of the abdomen and chest x-ray. Every 6 months computerized tomography of the abdomen and bone scans were performed. RESULTS: The overall 5-year survival rate was 69.1%, while the disease specific 5 year survival rate was 94%. One muscle invasive recurrence was noted at 69 months, which was again treated with the same regimen but ultimately led to radical cystectomy 21 months later. One patient died of progressive recurrence in the upper urinary tract. The 5-year recurrence-free survival rate was 46.5%. The only severe complication was BCG pneumonitis. CONCLUSIONS: The data show encouraging results for transurethral resection of bladder tumor with intravesical BCG therapy in select patients with T2a bladder cancer who are not candidates for radical cystectomy. PMID- 12576816 TI - Editorial: urothelial tumors--liberal applications of conservative approaches. PMID- 12576817 TI - Cyclooxygenase-2 is highly expressed in carcinoma in situ and T1 transitional cell carcinoma of the bladder. AB - PURPOSE: We describe cyclooxygenase-2 (COX-2) expression patterns in patients with carcinoma in situ and/or stage T1 transitional cell carcinoma. We determined whether expression is associated with clinical outcome in these patients. MATERIALS AND METHODS: Immunostaining for COX-2 was performed on paraffin embedded bladder biopsy specimens from 2 independent groups of patients without muscle invasive carcinoma, including 39 with carcinoma in situ and 34 with stage T1 tumors. Immunoreactivity was scored as the percent of carcinoma in situ cells with cytoplasmic staining for COX-2 in the carcinoma in situ group and as the percent of stage T1 cells expressing COX-2 in the stage T1 transitional cell carcinoma group. We evaluated other molecular alterations, including E-cadherin, p21 and p53, because evidence suggests a biological association of COX-2 with alterations in these molecular markers. RESULTS: In the carcinoma in situ group 5 patients (13%) had no immunoreactivity, while 2 (5%), 5 (13%) and 27 (69%) had 10%, 20% and 30% or greater carcinoma in situ cells positive for COX-2, respectively. In the transitional cell carcinoma group 1 (3%), 4 (12%) and 29 (85%) patients had 10%, 20% and 30% or greater positive cells, respectively. COX 2 expression was not associated with any clinical or pathological parameters, or with molecular markers regardless of the cutoff used. It was also not associated with clinical outcomes in the stage T1 transitional cell carcinoma group. In the carcinoma in situ group COX-2 expression was significantly associated with disease recurrence using cutoffs of 0% and greater than 10% positive cells, and with disease progression using a greater than 20% cutoff. However, it was not associated with bladder cancer related survival. CONCLUSIONS: COX-2 is expressed in a high percent of patients with carcinoma in situ and stage T1 transitional cell carcinoma, supporting the rationale for chemoprevention studies with selective COX-2. We could not substantiate a role for COX-2 immunohistochemistry for the staging and prognosis of carcinoma in situ and/or stage T1 transitional cell carcinoma. PMID- 12576818 TI - Superiority of ratio based lymph node staging for bladder cancer. AB - PURPOSE: The current study evaluated lymph node staging and the outcome in patients with lymph node positive bladder cancer after radical cystectomy. MATERIALS AND METHODS: A total of 162 patients with lymph node positive bladder cancer were followed a median of 7.5 years after radical cystectomy and pelvic lymph node dissection for survival and local recurrence. Lymph node disease was stratified by pN stage, the number of positive lymph nodes and the number of positive lymph nodes in relation to the number removed (ratio based pN stage). RESULTS: A median of 13 lymph nodes (range 2 to 32) was examined, showing an average of 3.3 positive lymph nodes per specimen. An increased number of lymph nodes correlated with the identification of lymph node positive cases. The ratio of the number of positive-to-total number of lymph nodes removed better defined surgical outcome than conventional lymph node staging. CONCLUSIONS: Ratio based lymph node staging, which reflects the number of lymph nodes examined and the quality of lymph node dissection, was a significant prognostic variable for survival and local control in patients with lymph node positive bladder cancer after radical cystectomy. PMID- 12576819 TI - Extent of pelvic lymphadenectomy and its impact on outcome in patients diagnosed with bladder cancer: analysis of data from the Surveillance, Epidemiology and End Results Program data base. AB - PURPOSE: The benefit of pelvic lymphadenectomy in patients with bladder cancer remains controversial. We analyzed the impact of lymphadenectomy on disease specific survival in a population based sample of patients with bladder cancer who underwent radical cystectomy. MATERIALS AND METHODS: Analysis included data on 1,923 patients who underwent radical cystectomy for bladder cancer between 1988 and 1996 obtained from the Surveillance, Epidemiology and End Results program cancer registry. We analyzed the impact of the number of lymph nodes examined, number of positive lymph nodes and ratio of positive-to-total number of lymph nodes resected on disease specific and overall survival independent of patient age, gender, stage, race, radiation and chemotherapy. RESULTS: Median followup in cystectomy cases was 63.5 months (range 0 to 131). Patients with 0 to 3 lymph nodes examined were at significantly higher risk of death from bladder cancer than those with greater than 3 (HR 1 to 1.2 versus 0.41 to 0.58). Patients with stages I/in situ, III and IV disease benefited from more extensive lymphadenectomy. In stage IV cases, while the total number of positive lymph nodes removed did not correlate with increased survival, the proportion of excised lymph nodes positive for metastatic bladder cancer tended to correlate with the risk of death from the disease. CONCLUSIONS: These results indicate significantly increased survival rates after cystectomy in patients with bladder cancer diagnosed with stages III or IV disease who have relatively more lymph nodes examined, suggesting that even some with higher stage disease may benefit from extended pelvic lymphadenectomy at cystectomy. PMID- 12576820 TI - Decreasing blood loss in patients treated with radical cystectomy: a prospective randomizes trial using a new stapling device. AB - PURPOSE: Radical cystectomy has been associated with significant blood loss and the need for heterologous transfusion. We investigated the potential decrease in blood loss and/or in transfusion requirement using a new stapling device compared with the traditional suture ligation technique. MATERIALS AND METHODS: We prospectively examined 70 patients with urothelial carcinoma who were scheduled for radical cystectomy. Each patient was randomized to traditional suture ligation or the Compact Flex Articulating Linear Cutter (Ethicon Endo-Surgery, Cincinnati, Ohio) stapling device. The 2 groups were prospectively compared with respect to estimated blood loss, transfusion requirement, operative time and complications. RESULTS: The groups were equivalent in terms of demographic and clinical variables, indicating that randomization produced 2 comparable groups. The stapler group had significantly lower estimated blood loss during cystectomy (p = 0.007) and during the whole procedure (p = 0.02). This group also required fewer transfusions (p = 0.006) and fewer mean units transfused (p = 0.003). The overall transfusion rate was 20% (14 of 70 cases). All patients in the stapler group had lower estimated blood loss and transfusion requirements. There was no statistical difference in time needed for bladder removal (p = 0.91) or total operative time (p = 0.17). No complications were attributable to the stapler device. CONCLUSIONS: In this prospective randomized study the stapling device significantly decreased blood loss and the transfusion requirement during radical cystectomy. These significant advantages combined with its relative safety make it an attractive surgical option and argue in favor of continued strategic attempts to decrease blood loss during radical cystectomy. PMID- 12576821 TI - Prognostic significance of vascular and perineural invasion in urothelial bladder cancer treated with radical cystectomy. AB - PURPOSE: Data on the prognostic significance of tumor invading lymphatic and blood vessels in bladder cancer are controversial, while little is known about perineural invasion in this tumor. We determined the prognostic value of these parameters in radical cystectomy specimens. MATERIALS AND METHODS: Slides of 283 radical cystectomy specimens obtained from 1986 to 1997 were examined retrospectively with respect to tumor invasion in lymphatic and blood vessels, and perineural spaces. This review was performed while blinded to lymph node tumor involvement or the postoperative disease course. The Kaplan-Meier probability analysis of tumor-free survival and the log rank test were used to determine the prognostic effects of vascular and perineural invasion. Multivariate analysis using the Cox proportional hazards model was also performed. RESULTS: Lymphatic, blood vessel and perineural tumor invasion were present in 54.1%, 13.1% and 47.7% of specimens, respectively. Tumor progressed in 46.3% of patients. On univariate analysis all 3 factors showed strong prognostic significance. However, on multivariate analysis only blood vessel invasion, invasion depth and regional lymph node status were independent prognostic factors (p <0.05). CONCLUSIONS: Lymph node metastases, pT classification and blood vessel invasion are independent prognostic parameters of tumor-free survival that should be used to guide patient treatment after radical cystectomy. Invasion of the blood and lymphatic vessels should be commented on separately in the pathology report. PMID- 12576822 TI - Value of urethral wash cytology in the retained male urethra after radical cystoprostatectomy. AB - PURPOSE: We determined the outcome in patients who underwent urethrectomy after cystectomy followed by routine urethral wash cytology versus those not followed by urethral wash cytology who presented with bleeding or urethral discharge. We retrospectively evaluated the outcome in post-cystectomy urethrectomy cases at our institution from 1994 to 2000. MATERIALS AND METHODS: A total of 24 patients with a median age of 70.5 years underwent urethrectomy after cystectomy, including 17 due to asymptomatic, positive urethral wash cytology (group 1) and 7 who were not followed by urethral wash and presented with bleeding/urethral discharge (group 2). Median time from cystectomy to urethrectomy was 11.4 months (range 6.7 to 67.1). Median followup after cystectomy and urethrectomy was 37 and 27.7 months, respectively. RESULTS: Urethrectomy pathological evaluation showed pTis disease in cases 12 (50%), pT0 in 9 (37.5%) and pT1 in 3 (12.5%). Cystectomy pathology was organ confined (pT0, pTis and pT1-pT2b disease) in 12 cases (50%), nonorgan confined (pT3a-pT4) in 6 (25%) and pT any N1 in 5 (21%). Cystectomy pathology was unknown in 1 case. At the most recent followup there was no evidence of disease in 14 patients (58%), 5 (21%) were alive with disease, 3 (12.5%) were dead of disease, 1 (4%) was dead of other causes and disease status was unknown in 1 (4%). There was no statistical difference in survival in groups 1 and 2 when controlling for original bladder tumor stage. Cox regression analysis revealed that cystectomy pathology was the only statistically significant parameter of disease-free survival (p = 0.011), while urethrectomy pathology and followup method (urethral washing versus no washing) were not significant. There were no perioperative or postoperative complications and no patients died. CONCLUSIONS: There was no significant survival difference in patients followed and not followed with urethral washing. Longer followup and increased patient numbers are needed to determine the significance of these findings. PMID- 12576823 TI - Do impalpable stage T1c prostate cancers visible on ultrasound differ from those not visible? AB - PURPOSE: We assessed whether the appearance of cancer within the prostate on sonography is associated with different pathological features and/or prognoses compared with nonvisible impalpable cancers defined as stage T1c by the TNM staging system. MATERIALS AND METHODS: We analyzed the clinical and pathological features, and progression rate in 323 patients with clinical stage T1cNX M0 cancer treated with radical prostatectomy between 1983 and 1998. Mean followup was 46.8 months (range 1 to 186). RESULTS: Of 323 impalpable stage T1c cancers 170 (53%) were visible and the remainder was not visible on ultrasound. There were no significant differences in clinical or pathological features of the cancers in these 2 groups. The prostate specific antigen nonprogression rate at 5 years was also similar for patients with impalpable cancer regardless of whether the lesion was or was not revealed by ultrasound (mean +/- SE 87% +/- 6% and 91% +/- 6%, respectively, p = 0.3767). Of the 170 visible cancers 55 patients had a hypoechoic lesion considered highly suspicious for cancer. These cancers were higher grade, more extensive, less likely to be confined to the prostate and the prognosis was significantly worse than that of impalpable cancer whether or not they were visible at a less suspicious level (IV or less, p = 0.011). However, such highly suspicious visible cancers are rarely visualized today. Initial serum prostate specific antigen more accurately predicts the pathological stage of impalpable cancer than transrectal ultrasound results. CONCLUSIONS: Impalpable cancers currently detected have similar pathological features and prognoses whether or not they are visible by ultrasound. Therefore, it is reasonable to categorize impalpable cancers as stage T1c and analyze the response to treatment regardless of the results of ultrasound. PMID- 12576824 TI - Comparison of preoperative prostate specific antigen density and prostate specific antigen for predicting recurrence after radical prostatectomy: results from the search data base. AB - PURPOSE: Prostate specific antigen (PSA) density based on the surgical weight of the radical prostatectomy specimen has previously been shown to be an independent predictor of biochemical recurrence after radical prostatectomy. We determined whether preoperative PSA density calculated using transrectal ultrasound prostate volume was a better predictor of advanced pathological findings or biochemical recurrence after radical prostatectomy relative to PSA. MATERIALS AND METHODS: We examined 552 men from the newly established Shared Equal Access Regional Cancer Hospital data base of men treated with radical prostatectomy at equal access medical centers to determine whether preoperative PSA density was a significant predictor of an adverse pathological condition or PSA recurrence after radical prostatectomy. Models using PSA density were compared with models using PSA to determine whether PSA density improved risk stratification relative to PSA. PSA density was examined as a continuous and a categorical variable using cutoffs to separate patients into groups at different risks for PSA failure. RESULTS: PSA density and PSA were significant predictors of adverse pathological findings on univariate analysis. Using PSA density in the multivariate model resulted in slightly better but statistically insignificant improvement in prediction of positive surgical margins (p = 0.134) and extracapsular extension (p = 0.771) relative to using PSA in the model. Neither PSA nor PSA density were significant independent predictors of seminal vesicle invasion. Area under the ROC curves for predicting biochemical recurrence for PSA and PSA density were not significantly different (0.589 and 0.58, respectively, p = 0.691). On separate multivariate analyses PSA density and PSA were significant independent predictors of biochemical failure. The multivariate model using PSA density provided only slight improvement in risk assessment relative to the model using PSA (index C = 0.589 and 0.581, respectively). To determine whether using PSA density as a categorical variable would result in improved prognostication we evaluated PSA density to determine the cutoff points that would provide the greatest risk stratification. PSA density cutoffs of less than 0.4, 0.4 to 1 and greater than 1 ng./ml./cc separated patients into 3 distinct groups at increasing risk for biochemical failure (p <0.001). While these cutoffs provided better risk stratification than when PSA density was examined as a continuous variable (index C = 0.684 versus 0.58), they provided only marginal improvement relative to the standard PSA cutoffs of less than 10, 10 to 20 and greater than 20 ng./ml. (index C = 0.676). CONCLUSIONS: The use of preoperative PSA density relative to PSA provided only slight improvement for predicting adverse pathological findings and biochemical recurrence after radical prostatectomy. The minimal and statistically insignificant improvement in preoperative risk assessment provided by PSA density does not justify the time and effort necessary to calculate this value. PMID- 12576825 TI - Conformal high dose rate brachytherapy improves biochemical control and cause specific survival in patients with prostate cancer and poor prognostic factors. AB - PURPOSE: To improve outcome for patients with prostate cancer with poor prognostic factors higher than conventional radiation doses are required. To achieve this outcome a brachytherapy boost was given. We report the results of the first high dose rate dose-escalation brachytherapy trial. MATERIALS AND METHODS: Between 1991 and 2000, 207 patients were prospectively enrolled in a dose escalation trial including pelvic radiotherapy and conformal high dose rate prostate brachytherapy boost. The dose was increased from 5.5 to 11.5 Gy. per implant. Patient eligibility for the study included pretreatment prostate specific antigen 10 or greater, Gleason 7 or greater or clinical stage T2b or higher. No patient received hormonal therapy. The American Society for Therapeutic Radiology and Oncology consensus panel definition of biochemical failure was applied. RESULTS: Median patient age was 69 years. Mean followup was 4.7 years (range 0.6 to 10.4). The 5-year actuarial biochemical control rate was 74%. The 5-year biochemical control was 85% for 1 poor prognostic factor, 75% for 2 and 50% for all 3 (p = 0.001). On Cox regression multivariate analysis lower brachytherapy dose, and higher Gleason and nadir value were associated with biochemical failure. The 5-year actuarial overall survival was 92%, cause specific survival 98% and disease-free survival 68%. The 5-year actuarial rates of complications were 8% and 0% for grades 3 and 4 genitourinary, and 0.5% and 0.5% for grades 3 and 4 gastrointestinal, respectively. The 5-year actuarial impotence rate was 51%. CONCLUSIONS: For patients with poor prognostic factors external beam radiation therapy with conformal high dose rate brachytherapy boost improved biochemical control, resulting in a high cause specific survival rate with low toxicity. Another important advantage is that the patient is not radioactive after the high dose rate implant. PMID- 12576826 TI - Corporoplasty for advanced Peyronie's disease using venous and/or dermis patch grafting: new surgical technique and long-term patient satisfaction. AB - PURPOSE: We evaluated long-term results and patient satisfaction after corporoplasty using saphenous vein and/or dermis patch grafting in patients with advanced Peyronie's disease. MATERIALS AND METHODS: A total of 20 patients who underwent venous and/or dermis patch corporoplasty were followed a mean of 19 months. Postoperatively patients satisfaction was assessed using the International Index of Erectile Function questionnaire. The mean preoperative grade of deviation in this collective was 86 degrees, frequently associated with additional malrotation. RESULTS: Intraoperatively the deviation was straightened in all cases. Postoperatively 20% of patients complained of 1 or 2 cm. penile shortening and in 25% there was a slight deviation relapse of 5 to 15 degrees. Only 2 patients had recurrence (30 and 70 degrees, respectively. In 3 cases (15%) postoperative erectile dysfunction was treated successfully with sildenafil or prostaglandin E. Overall postoperative sexual satisfaction score on the International Index of Erectile Function was 2.76 of a possible 5, although erection and cohabitation status were rated higher with at 3 and 2.94, respectively. CONCLUSIONS: Saphenous vein interposition is safe and effective for high grade Peyronie's disease. In cases of insufficient vein anatomy additional dermis patch interposition is a good alternative. The limited median postoperative patient satisfaction score even for minor procedures with good objective results indicates only moderate overall satisfaction at a mean of 19 months postoperatively, which was likely due to high preoperative patient expectation of surgical correction. PMID- 12576827 TI - Long-term outcome of ileal conduit diversion. AB - PURPOSE: Ileal conduit is considered a safe procedure and the gold standard to which newer forms of urinary diversion should be compared, although few long-term results are known. We analyzed a consecutive series of patients who lived a minimum of 5 years after ileal conduit diversion. MATERIALS AND METHODS: A total of 412 patients underwent ileal conduit diversion between 1971 and 1995 at our institution. We analyzed all conduit related complications occurring later than 3 months after surgery in 131 long-term survivors (survival 5 years or greater). RESULTS: Median followup was 98 months (range 60 to 354). Overall 192 conduit related complications developed in 87 of 131 (66%) patients. The most frequent complications were related to kidney function/morphology in 35 patients (27%), stoma in 32 (24%), bowel in 32 (24%), symptomatic urinary tract infection (including pyelonephritis) in 30 (23%), conduit/ureteral anastomosis in 18 (14%) and urolithiasis in 12 (9%). Within the first 5 years complications developed in 45% of patients. This percentage increased to 50%, 54% and 94% in those surviving 10, 15 and longer than 15 years, respectively. In this last group 50% had upper urinary tract changes and 38% had urolithiasis, for which the respective numbers after 5 years were 12% and 17%. CONCLUSIONS: This study demonstrates a high conduit related complication rate in long-term survivors and underlines the need for vigorous long-term followup. Only studies lasting more than 1 decade cover the entire morbidity spectrum. PMID- 12576828 TI - Comparison of open surgery versus retroperitoneoscopic approach to chyluria. AB - PURPOSE: We compared the clinical effectiveness of renal pedicle lymphatic disconnection for chyluria performed by retroperitoneoscopy and by open surgery. MATERIALS AND METHODS: Three male and 4 female patients 33 to 68 years old (mean age 49) with chyluria underwent retroperitoneoscopic renal pedicle lymphatic disconnection. Chyluria was on the left side in 5 cases and on the right side in 2. Open renal pedicle lymphatic disconnection was performed in 4 men and 2 women 33 to 61 years old (mean age 45.8). Chyluria was on the left and right sides in 3 cases each. Mean operative time, intraoperative blood loss, postoperative intestinal function recovery time, intraoperative and postoperative complications, postoperative hospital delay and operative outcome were compared in these 2 groups. RESULTS: Compared with the open surgery group results in the retroperitoneoscopic group were superior in terms of operative time (42 to 90 minutes, mean +/- SD 65.0 +/- 18.8 versus 120 to 220, mean 156.7 +/- 38.8), intraoperative blood loss (20 to 50 ml., mean 29.3 +/- 10.2 versus 60 to 250, mean 171.7 +/- 76.5), postoperative intestinal function recovery time (24 to 48 hours, mean 36.0 +/- 6.9 versus 24 to 72, mean 54.0 +/- 21.1), intraoperative and postoperative complications, and postoperative hospital stay (3 to 6 days, mean 4.7 +/- 0.7 versus 7 to 9 days, mean 7.8 +/- 1.0). In the open surgery group primary anastomosis was performed in 1 case due to injury to a renal artery branch during the operation. Chyluria resolved the day after surgery in the 2 groups. No obvious complications developed postoperatively. The followup of 2 to 12 months (mean 6.7 +/- 4.0) showed no recurrence of chyluria. CONCLUSIONS: Retroperitoneoscopic renal pedicle lymphatic disconnection completely ligates the lymphatic vessels with minimal invasion, less blood loss, rapid recovery and a good short-term outcome. PMID- 12576829 TI - A prospective randomized trial of the LoFric hydrophilic coated catheter versus conventional plastic catheter for clean intermittent catheterization. AB - PURPOSE: We compared the incidence of hematuria, pyuria and clinical urinary tract infection in patients who performed intermittent self-catheterization using a hydrophilic coated LoFric (Astra Tech AB, Molndal, Sweden) or standard plastic catheter. MATERIALS AND METHODS: A total of 62 male patients who performed intermittent self-catheterization to manage neurogenic bladder were randomized into 2 treatment groups at 3 American study sites. Outcome measures included urinary tract infection, microhematuria, pyuria and satisfaction rates. RESULTS: Of the 62 enrolled patients 49 completed the 12-month study. The withdrawal rate was not different in the 2 groups. At the end of the study there was statistically significantly less hematuria in the hydrophilic coated catheter group compared with controls. In addition, there was a significant decrease in the urinary tract infection rate from baseline in the hydrophilic coated catheter group but not in controls. CONCLUSIONS: Use of the hydrophilic coated catheter by patients on intermittent self-catheterization is associated with less hematuria and a significant decrease in the incidence of urinary tract infections. Therefore, it may be preferable for some patients, especially those with a history of difficult catheterization, urethral trauma or a high rate of urinary tract infection. PMID- 12576830 TI - Sphincteric urinary incontinence: relationship of vesical leak point pressure, urethral mobility and severity of incontinence. AB - PURPOSE: We examined the relationships among urethral hypermobility, intrinsic sphincter deficiency and incontinence in women. MATERIALS AND METHODS: A total of 65 consecutive women with stress urinary incontinence and 28 with lower urinary tract symptoms not associated with stress urinary incontinence were evaluated with videourodynamics, 24-hour voiding diaries and pad tests, vesical leak point pressure measurement and the cotton swab test. RESULTS: A total of 93 patients with a mean age +/- SD of 63 +/- 13 years were studied, including 65 who presented with stress urinary incontinence and 28 who presented with lower urinary tract symptoms without stress urinary incontinence. The incidence of urethral hypermobility was 32% in the stress urinary incontinence group and 36% in the lower urinary tract symptoms group (p = 0.46). When stress urinary incontinence cases were stratified according to a vesical leak point pressure of 0 to 60, 60 to 90 and greater than 90 cm. H2O, urethral hypermobility was noted in 25%, 31% and 41%, respectively, a difference that was not statistically significant (p = 0.6). Overall incontinent patients with and without urethral hypermobility had the same median number of incontinence episodes (5, range 1 to 13 versus 7, range 1 to 15, p = 0.39) and median pad weight (39.5 range 1 to 693 gm. versus 33.5, range 1 to 751, p = 0.19). When patients with intrinsic sphincter deficiency, defined as vesical leak point pressure less than 60 cm. H2O, were divided into those with and without urethral hypermobility, there were no differences in the mean number of incontinence episodes (9.4 +/- 3 versus 6 +/ 3.6, p = 0.17) or median pad weight (90 gm., range 10 to 348 versus 86, range 30 to 81, p = 0.76). The degree of change in the urethral angle did not correlate with vesical leak point pressure (r = 0.16, p = 0.24) or with pad weight (r = 0.23, p = 0.1). CONCLUSIONS: Urethral hypermobility was equally common in this group of women with lower urinary tract symptoms and stress urinary incontinence. Intrinsic sphincteric deficiency and urethral hypermobility may coexist and they do not define discrete classes of patients with stress urinary incontinence. Urethral hypermobility did not appear to have an independent effect on the frequency or severity of incontinence. Patients with stress urinary incontinence can still be characterized by vesical leak point pressure and change in the urethral angle, although these variables do not always define discrete classes. PMID- 12576831 TI - Noninvasive measurement of bladder pressure. Does mechanical interruption of the urinary stream inhibit detrusor contraction? AB - PURPOSE: As part of developing a noninvasive method to measure bladder pressure using an inflatable penile cuff, we tested the hypothesis that detrusor contraction is maintained without inhibition during the test. MATERIALS AND METHODS: Five healthy volunteers and 26 male patients with lower urinary tract symptoms underwent interruption of established urine flow by controlled inflation of a cuff placed around the penis with simultaneous invasive bladder pressure monitoring. After interruption of flow the cuff was rapidly deflated and voiding was allowed to resume. The bladder pressure was recorded before, during and after interruption of flow by cuff inflation. RESULTS: During flow interruption an isovolumetric increase in detrusor pressure was observed. When the cuff was deflated the detrusor pressure quickly returned to preinflation values and urine flow immediately resumed. Intra-abdominal pressure did not change during the cuff inflation cycle. CONCLUSIONS: Mechanical interruption of urine flow by controlled inflation of a penile cuff during voiding does not inhibit detrusor contraction. This finding further validates our noninvasive technique of bladder pressure measurement and supports ongoing studies into its clinical usefulness. PMID- 12576832 TI - Bladder outlet resistance. AB - PURPOSE: We developed a method to objectively compare methods for the quantification of bladder outlet resistance. MATERIALS AND METHODS: We studied unselected voiding cystometries from 131 male patients. Several models proposed for bladder outlet resistance were fitted to the lowest monotonically increasing part (bottom) of the pressure flow plots. In conjunction with a parameter for bladder contractility, the model parameters were used as predictors of post-void residual greater than 20% of filled volume. RESULTS: The pressure decrease in relaxed bladder outlet was best described by a linear function of the flow rate. The flow rate independent and dependent parts of that function were statistically significant predictors for residual. However, they could be combined into 1 index that was as efficient as the 2 numbers separately in predicting residual. This index is the average pressure of the bottom of the pressure flow plot. CONCLUSIONS: We describe how different models for the quantification of bladder outlet resistance can be compared objectively on the basis of their ability to predict a significant post-void residual. Using this criterion in an unselected group of measurements, the average pressure of the bottom of the pressure flow plot performed best as an index for bladder outlet resistance quantification. It combines (or weights) 2 components (dependent and independent flow rates) without losing its power to predict post-void residual. PMID- 12576833 TI - Editorial: bladder failure--a condition to reckon with. PMID- 12576834 TI - Repeat urethrolysis after failed urethrolysis for iatrogenic obstruction. AB - PURPOSE: Bladder outlet obstruction is a potential complication of all stress incontinence surgery. Urethrolysis successfully relieves 65% to 93% of cases. We determined the success of repeat urethrolysis after failed initial urethrolysis to relieve obstruction. MATERIALS AND METHODS: We reviewed the charts of 24 women who underwent repeat urethrolysis for iatrogenic obstruction after at least 1 previous attempt. Aggressive repeat urethrolysis was performed via a retropubic or transvaginal route depending on the clinical scenario and surgeon discretion. Outcomes measured were patient ability to void spontaneously without catheterization, decreased post-void residual urine and resolution of lower urinary tract symptoms, particularly urge incontinence. RESULTS: Mean patient age was 55 years (range 38 to 80). The initial incontinence procedure was a pubovaginal sling in 10 cases, retropubic suspension in 9, needle suspension in 4 and anterior colporrhaphy in 1. A total of 23 patients had previously undergone transvaginal urethrolysis, while 1 had undergone retropubic urethrolysis. The repeat procedure was retropubic in 12 women (50%), transvaginal in 10 (42%) and combined in 2 (8%). Mean time between initial and repeat urethrolysis was 9 months (range 1 to 3). Mean followup was 14 months. Postoperatively 20 of the 22 patients (91%) who were catheter dependent no longer needed to catheterize. Post void residual urine normalized in the 2 patients who had not been catheter dependent but who had had elevated post-void residual urine. Thus, repeat urethrolysis successfully eliminated urinary retention in 22 of the 24 cases (92%). Mean post-void residual urine before and after repeat urethrolysis was 334 versus 44 ml. (p <0.001). Irritative symptoms and urge incontinence completely resolved in 12% of cases, were improved and required medication in 69% and remained the same in 19%. No patient had new onset irritative symptoms. Stress urinary incontinence recurred in 4 of the 22 women (18%) and persisted in the 2 in whom it had been present before urethrolysis. CONCLUSIONS: Aggressive repeat urethrolysis can be highly successful for relieving iatrogenic retention. Complete resolution of irritative symptoms and urge incontinence is less likely. Recurrent stress urinary incontinence is similar to that after primary urethrolysis. PMID- 12576835 TI - Effect of urethral dilation on vesical motor activity: identification of the urethrovesical reflex and its role in voiding. AB - PURPOSE: We investigated the hypothesis that mild urethral distention, which presumably occurs during the passage of urine through the urethra, stimulates stretch receptors in the urethral wall, leading reflexively to vesical contraction. MATERIALS AND METHODS: We evaluated 9 male and 10 female healthy volunteers with a mean age +/- SD of 39.6 +/- 8.3 years. The posterior urethra was distended by a balloon filled with saline in 1 ml. increments up to 6 ml., while recording vesical pressure. The test was repeated after individual anesthetization of the urethra and bladder. RESULTS: Vesical pressure increased significantly at 1 and 2 ml. urethral distention (p <0.01). Increases in urethral distention effected further vesical pressure elevation (p <0.001), although there was no significant difference in distention at 3 to 6 ml. (p >0.05). No significant vesical pressure response of the individually anesthetized urethra or bladder occurred during urethral distention. CONCLUSIONS: Urethral distention is thought to cause vesical contraction through the stimulation of urethral stretch receptors. Vesical contraction at urethral distention postulates a reflex relationship that was abolished by individual anesthetization of the urethra and bladder. This relationship, which we call the urethrovesical reflex, appears to have a role in maintaining vesical contraction during voiding. Further studies are required to investigate the role of this reflex in voiding disorders. PMID- 12576836 TI - A new endoscopic ureteral reimplantation for primary vesicoureteral reflux (endoscopic trigonoplasty II). AB - PURPOSE: We describe a new technique of endoscopic antireflux surgery. The principle of the procedure is to make a reliable muscular backing and elongate the intramural ureter. MATERIALS AND METHODS: We performed this new endoscopic surgery in 8 female patients in whom 4, 1, 8 and 1 refluxing ureters (total 14) were diagnosed with grades I to IV reflux, respectively. The operation consists of 3 steps. Two 5 mm. locking trocars are placed into the bladder. Irrigation is done with 3% D-sorbitol solution and the bladder wall is incised upward along each side of the ureter using a resectoscope to make a 2 to 3 cm. U-shaped bladder flap, including the ureter. Under pneumobladder the incised muscle is sutured to make a muscular bed with a needle holder via the urethra and forceps via the abdominal trocar. The U flap is fixed with 2 distal anchor sutures on the embedded muscular layer and 4 additional sutures are placed to approximate the mucosa of the U-shaped flap and bladder. RESULTS: Mean operative time was 245 minutes. Ureteral injury occurred in 2 patients. A Foley catheter remained indwelling for 3 to 5 days (mean 4.1). Reflux resolved in 12 of the 14 ureters (86%) 12 months postoperatively. Vesicoureteral reflux persisted in 1 case because of insufficient fixation and recurred in 1 because of ureterovesical fistula. The patients were satisfied with better cosmesis and minimal postoperative discomfort. CONCLUSIONS: We believe that procedure is feasible for female patients with primary vesicoureteral reflux. PMID- 12576837 TI - Which stop test is best? Measuring detrusor contractility in older females. AB - PURPOSE: Impaired detrusor contractility has an important role in geriatric voiding dysfunction but there are many competing methods of measurement. We compared the performance of 3 methods of measuring detrusor contraction strength to identify the best one. MATERIALS AND METHODS: We retrospectively analyzed urodynamics data on 84 females 53 years old or older. All had urge incontinence and were enrolled in a placebo controlled oxybutynin trial. Stop tests (voluntary interruption, mechanical interruption and continuous mechanical occlusion of flow) were performed on each subject. RESULTS: At baseline the voluntary stop test measured lower mean isovolumetric detrusor pressure +/- SD than the mechanical and continuous methods (31.2 +/- 16.0 versus 47.2 +/- 26.5 and 48.7 +/ 24.4 cm. water, respectively). The latter 2 values also correlated highly (r = 0.87). Followup data on 76 women confirmed these results. Based on baseline and followup values in the 20 women who received placebo the continuous occlusion test showed highest test-retest reliability (r = 0.9, p <0.01), followed by the mechanical (r = 0.69, p = 0.01) and voluntary (r = 0.67, p <0.01) stop tests. Treatment with oxybutynin decreased isovolumetric detrusor pressure in all 3 stop tests by up to 6 cm. water. However, the decrease was statistically significant only for the continuous occlusion test. CONCLUSIONS: To assess detrusor contraction strength in elderly females with urge incontinence the mechanical stop and continuous occlusion tests are acceptable but the continuous occlusion test has better reliability and better detects slight drug induced changes. Voluntary stop tests greatly underestimate detrusor isovolumetric pressure and should no longer be used. PMID- 12576838 TI - Transvaginal radio frequency treatment of the endopelvic fascia: a prospective evaluation for the treatment of genuine stress urinary incontinence. AB - PURPOSE: We evaluate the safety and efficacy of a new treatment modality for genuine stress urinary incontinence which was a transvaginal radio frequency applicator to deliver radio frequency energy to the endopelvic fascia. The purported mechanism of effect for this therapy is shrinkage of the collagenated tissue which composes the endopelvic fascia that supports the bladder neck and proximal urethra, thus stabilizing the proximal urethra and bladder neck. In prior animal trials and early pilot studies this therapy was shown to cause a reproducible thermal effect manifested by fascial shrinkage. Preliminary human trials indicated a therapeutic benefit of this therapy for women with genuine stress urinary incontinence. MATERIALS AND METHODS: To our knowledge this is the first multicenter study of a transvaginal approach for radio frequency of the endopelvic fascia for treatment of genuine stress incontinence. Between June 1999 and June 2000, 120 consecutive women (mean age 49.9 years) at 10 sites underwent transvaginal radio frequency treatment in a prospective trial to evaluate the overall efficacy and safety profile of this therapy. All patients had preoperative urethral hypermobility (average cotton swab change 38 degrees). Detrusor instability was excluded by cystometry. In all procedures precisely controlled radio frequency energy was applied to the endopelvic fascia to heat and shrink the tissue. The patients were evaluated postoperatively at 1 week and at 1, 3, 6 and 12 months using objective and subjective measures. Primary end points consisted of physician assessment of continence, patient reported pad use and the number of patient reported episodes. Safety was determined for acute (immediate postoperative) and chronic time frames. RESULTS: Of the 120 patients 96 completed 1-year evaluation. Average operative time was less than 30 minutes, and all patients were treated as outpatients. Preoperatively 101 patients (84%) averaged 1 or more episodes of urinary incontinence per day. At 3, 6 and 12 months 57%, 66% and 59% of patients, respectively, averaged 1 or no daily episodes of urinary incontinence. At 12-month followup 79 of 109 patients (73%) reported being continent or improved. Preoperatively, 43% of patients reported using 1 or no pads daily. At 3, 6 and 12 months 69%, 70% and 72% of patients, respectively, required 1 or no pads daily. On urodynamic evaluation at 12-month followup 76.0% of the patients did not leak with a Valsalva maneuver. A total of 30 cases were classified as failures and 11 women were lost to followup. There were no intraoperative complications, 3 (4%) minor postoperative complications which resolved, and no device related complications. CONCLUSIONS: The transvaginal radio frequency applicator demonstrated good efficacy and excellent safety at 1-year followup. Ongoing analysis of the data has indicated opportunities for improvement of this new surgical technique that could result in higher efficacy rates without compromising safety. Further long-term evaluation is being conducted to assess chronic durability of the procedure. PMID- 12576839 TI - Ten-year experience with transvaginal vesicovaginal fistula repair using tissue interposition. AB - PURPOSE: The success rate of vesicovaginal fistula repair is improved by tissue interposition. The Martius flap produces reliable results but it has increased morbidity. A peritoneal flap is easily created with minimal morbidity and it can be used for proximal fistulas. We describe our 10-year experience with tissue interposition for transvaginal repair of vesicovaginal fistulas. MATERIALS AND METHODS: From January 1991 to July 2001, 207 cases of vesicovaginal fistulas were repaired transvaginally. Tissue interposition was used for complex (greater than 2 cm. and/or radiation induced) fistulas and/or failed previous repairs. A peritoneal flap was used for proximal fistulas and a Martius flap was used for distal fistulas. A full-thickness labial flap was reserved for cases of insufficient vaginal epithelium. RESULTS: A total of 207 patients underwent transvaginal repair of a vesicovaginal fistula. Etiology of the fistula was hysterectomy in 91% of cases (abdominal in 83% and vaginal in 8%), radiation in 4% and 5% other (obstetric trauma, anterior colporrhaphy or an indwelling catheter) in 5%. In 159 patients (77%) at least 1 previous repair had failed. Repair in 120 patients (58%) was done with tissue interposition, including a peritoneal, Martius and full-thickness labial flap in 83, 34 and 3, respectively. The cure rate after initial repair with a peritoneal, Martius and labial flap was 96%, 97% and 33%, respectively. There were no intraoperative complications. CONCLUSIONS: A peritoneal flap for transvaginal repair of vesicovaginal fistulas has minimal morbidity, results in a success rate comparable to that of the Martius flap and is especially useful for proximal fistulas when previous repair has failed. PMID- 12576840 TI - A prospective, randomized pilot trial of acupuncture of the kidney-bladder distinct meridian for lower urinary tract symptoms. AB - PURPOSE: We performed a pilot trial to assess the response of lower urinary tract symptoms and prostate specific antigen (PSA) to acupuncture in a population of patients biopsy negative for prostate cancer. MATERIALS AND METHODS: A total of 30 patients were randomly assigned to 1 of 3 study groups, including observation for 3 months with 6 blood samples for PSA at set intervals, 9 sessions of acupuncture in 3 months to points of the kidney-bladder distinct meridian expected to treat the prostate with 6 blood samples for PSA at set intervals and 9 sessions of acupuncture in 3 months to points not expected to treat the prostate with 6 blood samples for PSA at set intervals. The effect of acupuncture on lower urinary tract symptoms was assessed monthly using the International Prostate Symptom Score. RESULTS: Trend analysis (repeated measures ANOVA) revealed no significant changes in the 3-month period in the randomized arms. Statistical analysis showed p = 0.063 for the International Prostate Symptom Score, p = 0.945 for PSA and p = 0.37 for the free-to-total PSA ratio. CONCLUSIONS: Acupuncture to the kidney-bladder distinct meridian neither relieves lower urinary tract symptoms nor impacts PSA. PMID- 12576841 TI - Use of medications for erectile dysfunction in the United States, 1996 through 2001. AB - PURPOSE: We describe the use during 1996 through 2001 of the primary medications approved in the United States for treatment of erectile dysfunction, namely alprostadil injection and urethral suppository, and sildenafil. MATERIALS AND METHODS Two pharmaceutical research data bases, the National Prescription Audit Plus, and National Disease and Therapeutic Index, were accessed and analyzed. Ancillary data were obtained from 2 health plans. RESULTS: Increases in the number of dispensed prescriptions for alprostadil injection and urethral suppository marketed in 1995 and 1996, respectively, were reversed in 1998 by the marketing of sildenafil. From 1998 through 2001 the estimated number of prescriptions for sildenafil increased 1.87-fold or 87% from 7.5 million to 14 million, while those for alprostadil injection decreased 33% from 239,000 to 159,000 and those for alprostadil suppository decreased 67% from 400,000 to 132,000. Sildenafil was prescribed proportionately more frequently for younger men than alprostadil injection or suppository (p <0.0001). Compared with men for whom sildenafil was prescribed in 1998 those prescribed the drug in 2001 were younger (p <0.0001). Alprostadil injection and suppository were prescribed proportionately more frequently by urologists than sildenafil. Ancillary data from 2 health plans indicated a 173% increase in 1 plan and a 25% decrease in the other due to restrictions in sildenafil prescriptions from 1998 through 2001. CONCLUSIONS: Due to the marketing of sildenafil in 1998 through 2001 the use of 2 approved medications for erectile dysfunction, namely alprostadil injection and alprostadil urethral suppository, decreased, while the use of sildenafil increased. Sildenafil was prescribed proportionately more frequently for younger men than alprostadil injection or suppository. Alprostadil was prescribed proportionately more frequently by urologists than sildenafil, which was most commonly prescribed by family and general practitioners, and internists. The data indicate the wide adoption and use of sildenafil for erectile dysfunction. PMID- 12576842 TI - Erectile dysfunction risk factors in noninsulin dependent diabetic Saudi patients. AB - PURPOSE: We assessed the prevalence of and analyzed risk factors for erectile dysfunction in patients with noninsulin dependent diabetes in Makkah, Saudi Arabia. MATERIALS AND METHODS: A total of 562 male diabetic Saudi patients were enrolled in this study. Patients were screened for erectile dysfunction using the International Index of Erectile Function. At the time of screening patients were also interviewed for sociodemographic data, including age, education, occupation, marital status and smoking. Medical history included diabetes, diabetes related complications, risk factors for diabetes and erectile dysfunction, and current medication. RESULTS: Mean age of the study sample was 53.7 years (range 27 to 84). Of the patients 86.1% had various degrees of erectile dysfunction, including mild in 7.7%, moderate in 29.4% and severe in 49.1%. The prevalence of erectile dysfunction was 25% in patients younger than 50 years, which increased to 75% in those older than 50 years. Of those without erectile dysfunction 70% were younger and 30% were older than 50 years (p = 0.0001). Patients with a history of diabetes of greater than 10 years were 3 times as likely to report erectile dysfunction as those with a history of less than 5 years. Men with poor metabolic control were 12.2 times as likely to report erectile dysfunction as those with good metabolic control. Of diabetic patients with erectile dysfunction 53% had 1 or more diabetic related complications compared with 20.5% with no erectile dysfunction (p = 0.0001). CONCLUSIONS: Erectile dysfunction is common in diabetic Saudi men. This study provides a quantitative estimate of the prevalence of erectile dysfunction and its main risk factors in diabetic Saudi patients. PMID- 12576843 TI - Sildenafil response is influenced by the G protein beta 3 subunit GNB3 C825T polymorphism: a pilot study. AB - PURPOSE: Sildenafil is the oral phosphodiesterase-5 inhibitor that revolutionized treatment for erectile dysfunction. We investigated a potential association of the G protein beta 3 subunit (GNB3) C825T polymorphism, a determinant of intracellular signal transduction, with the drug response to sildenafil in patients with erectile dysfunction. MATERIALS AND METHODS: In 113 men with erectile dysfunction and 111 healthy male controls genotype status of the GNB3 C825T polymorphism was determined by polymerase chain reaction and restriction analysis. Patients with erectile dysfunction received sildenafil at a dose of 25 to 100 mg. according to the individual erectile response. Drug response was measured by interviewing the patient according to the erection scale of 0 to 5. RESULTS: The GNB3 genotype distribution of patients with erectile dysfunction exactly matched that of healthy controls. Analysis of the response to sildenafil revealed a significant association of C825T allele status with the erectile response to sildenafil. In the group with TT genotype we observed a 90.9% response but only a 50.9% and 48.9% response in patients with the CC and TC genotype, respectively. The odds ratio for a positive erectile response was 10.0 (95% CI 1.2 to 81.1) for patients with the TT versus the TC/CC genotype (p = 0.01). CONCLUSIONS: The response to sildenafil is significantly associated with GNB3 C825T genotype status in patients with erectile dysfunction. PMID- 12576844 TI - Microsurgical vasovasostomy after failed vasovasostomy. AB - PURPOSE: We analyzed our experience with repeat microsurgical vasovasostomy after failed vasovasostomy and elucidate the possible predictors of surgical outcome. MATERIALS AND METHODS: We evaluated 62 repeat vasectomy reversal cases with followup data available. Regardless of the intraoperative observation of sperm in the vasal fluid bilateral microsurgical 2-layer vasovasostomy was performed when surgically possible. Of these 62 patients 60 (97%) underwent bilateral (58) or unilateral (2) vasovasostomy and 2 (3%) underwent unilateral vasovasostomy with contralateral epididymovasostomy. RESULTS: Patency and pregnancy followup data were available on 62 and 42 patients, respectively. The overall patency and pregnancy rates achieved were 92% and 57%, respectively, and the natural birth rate was 52%. Increased age of the wife proved a negative prognostic factor for pregnancy (p = 0.018). The intraoperative detection of sperm and other factors, including obstructive interval, reconstruction type, anastomotic site, patient age and postoperative semen parameters, did not influence the surgical outcome. CONCLUSIONS: Regardless of the detection of sperm in the intravasal fluid during the operation repeat microsurgical vasovasostomy resulted in a better outcome than in other studies, in which adopted epididymovasostomy was done when sperm was absent from the vas fluid. Our study suggests that compromised anastomosis after previous surgery is the most common cause of failed vasovasostomy. We recommend that microsurgical vasovasostomy should be performed preferentially in failed vasovasostomy cases. PMID- 12576845 TI - Comparative study of sclerotherapy with phenol and surgical treatment for hydrocele. AB - PURPOSE: We compared the efficiency, side effects and effects on spermatogenesis of sclerotherapy with phenol and surgical treatment for hydrocele. MATERIALS AND METHODS: A total of 67 patients (80 hydroceles) were randomly divided into 2 groups of 40 hydroceles each. One group underwent phenol sclerotherapy and the other underwent hydrocelectomy. Spermiograms were done before, 6 and 12 months after treatment in patients able to ejaculate. RESULTS: In the sclerotherapy group 47.5%, 30%, 12.5%, 5% and 2.5% of the hydroceles were cured with 1 to 5 injections, respectively, but 2.5% were not cured even with 6 injections. There were no complaints of localized pain or infection in these cases. All patients returned to normal activity on the same day. In the hydrocelectomy group 97.5% and 2.5% of hydroceles were cured with 1 and 2 operations, respectively. There was pain postoperatively in 73.5% of the patients and localized infection in 5%, while 62.5% required an average of 4.5 days of rest and were absent from work for 10 days. There was no significant statistical alteration in spermatozoid concentration in the preoperative, and 6 and 12-month postoperative counts in the groups (p = 0.385). CONCLUSIONS: Sclerotherapy for hydrocele with phenol is as efficacious as hydrocelectomy, while causing less morbidity and similar effects on spermatogenesis. PMID- 12576846 TI - Ureteral stent symptom questionnaire: development and validation of a multidimensional quality of life measure. AB - PURPOSE: We developed the ureteral stent symptom questionnaire (USSQ), a psychometrically valid measure to evaluate symptoms and impact on quality of life of ureteral stents. MATERIALS AND METHODS: A total of 309 patients were asked to participate during different phases of our study. In phase 1 a structured literature search, 9 patient interviews and studies of 90 patients using existing instruments formed the foundation for the initial draft of our new questionnaire. In phase 2 the USSQ was pilot tested, reviewed by experts and field tested in 40 patients to produce a final 38-item draft. In phase 3 formal validation studies were performed in 55 patients to assess validity, reliability and sensitivity to change. Discriminant validation was performed by administering the questionnaire to 3 groups of patients without stents. RESULTS: The final draft addressed various domains of health (6 sections and 38 items) affected by stents covering urinary symptoms, pain, general health, work performance, sexual matters and additional problems. The validation studies showed the questionnaire to be internally consistent (Cronbach's alpha > 0.7) with good test-retest reliability (Pearson's coefficient > 0.84). The questionnaire demonstrated good construct validity and sensitivity to change shown by significant changes in the score with and after removal of stents. The new USSQ discriminated patients with stents from healthy controls (p < 0.001) and patients with urinary calculi without stents and lower urinary tract symptoms. CONCLUSIONS: Indwelling ureteral stents have a significant impact on health related quality of life. The new USSQ is a valid and reliable instrument that is expected to become a standard outcome measure to evaluate the impact and compare different types of stents. PMID- 12576847 TI - Indwelling ureteral stents: evaluation of symptoms, quality of life and utility. AB - PURPOSE: We report the prevalence of symptoms associated with ureteral stents, their impact on health related quality of life and utility analysis based on the validation studies of the new ureteral stent symptom questionnaire (USSQ). MATERIALS AND METHODS: A total of 85 consecutive adult patients with unilateral indwelling ureteral stents who were asked to participate during the validation phases of the USSQ were considered for this analysis. They were asked to complete the USSQ and the EuroQol, a weighted utility instrument, 4 weeks after stent insertion and removal. In addition, 40 patients were asked to complete these questionnaires 1 week after stent insertion to assess the prevalence of symptoms and utility values at different times. RESULTS: Of the 85 patients 62 (73%) with a mean age of 50 years completed the necessary questionnaires. Urinary symptoms and pain that affected work performance and general health were important stent related problems. Of the patients 78% reported bothersome urinary symptoms that included storage symptoms, incontinence and hematuria. More than 80% of patients experienced stent related pain affecting daily activities, 32% reported sexual dysfunction, and 58% reported reduced work capacity and negative economic impact. The mean EuroQol utility values, which indicate patient satisfaction with treatment, were significantly reduced following stent insertion. CONCLUSION: Urinary symptoms and pain associated with indwelling ureteral stents interfere with daily activities and result in reduced quality of life in up to 80% of patients. Stents are associated with negative functional capacity and reduced utility values. The results have implications in terms of routine clinical practice, patient counseling and future stent research. PMID- 12576848 TI - Ureteral access sheath facilitates inspection of incision of ureteropelvic junction. AB - PURPOSE: The Acucise (Applied Medical, Rancho Santa Margarita, California) electrocautery balloon is a highly successful device used in managing congenital and secondary ureteropelvic junction obstruction. Correct orientation of the cutting wire is essential during insertion of the Acucise catheter to avoid injury to crossing vessels. Moreover, confirmation of the lateral ureteropelvic junction incision is typically verified by fluoroscopic identification of extravasated contrast material. We describe a technique of facilitated passage of the Acucise balloon through a ureteral access sheath followed by ureteroscopic visualization of the incision, affording the opportunity to improve the incision with the holmium laser if necessary. MATERIALS AND METHODS: After retrograde pyelography and guidewire placement, a 12/14Fr, 35 cm. ureteral access sheath is fluoroscopically introduced to the proximal ureter. The Acucise balloon is advanced across the ureteropelvic junction and the balloon is partially inflated to confirm proper placement. Following lateral Acucise incision, flexible ureteroscopy allows direct visualization of the ureteropelvic junction, confirming a through-and-through incision. Completion of a partial incision can be performed if needed with a 200 micro holmium laser fiber followed by routine stent placement. RESULTS: During the last 8 months we have used the Acucise device through a ureteral access sheath to treat congenital or secondary ureteropelvic junction obstruction in 8 patients. All incisions demonstrated extravasation of contrast material on retrograde pyelography, and 6 incisions (75%) were noted to be transmural by flexible ureteroscopic inspection. Two patients (25%) with only a partial incision despite contrast extravasation underwent extended incision using the holmium laser. Short-term followup demonstrated patency of the ureteropelvic junction in 7 of the 8 patients (87.5%) with 1 eventually requiring a secondary open pyeloplasty. CONCLUSIONS: The ureteral access sheath greatly facilitates placement of the Acucise device and allows rapid ureteroscopic confirmation of the incision. Insertion and removal of the ureteral access sheath and flexible ureteroscope do not compromise or significantly increase the duration of the procedure. Moreover, flexible ureteroscopic visualization allows confirmation of a complete transmural incision and potentially increases success rates of this minimally invasive approach to ureteropelvic junction obstruction. Continued followup is necessary to confirm the long-term benefits of this procedure. PMID- 12576849 TI - Bridging long ureteral defects using the Yang-Monti principle. AB - PURPOSE: A new technique for replacing the ureter based on the Yang-Monti principle is introduced to overcome the drawbacks of the classic ileal ureter. MATERIALS AND METHODS: Between March 2001 and June 2002 ureteral replacement by ileum was indicated in 10 patients, including 9 with long or multiple strictures due to bilharzial (6), tuberculosis (3) and a low grade lower ureteral tumor (1). The technique involved isolation of a 5 to 7.5 cm. ileal segment, which was further subdivided into 2 or 3 equal parts. Paramesenteric incision along the longitudinal axis of these segments followed by unfolding resulted in a 12 to 18 cm. ileal strip. Tubularization of this strip led to the formation of an ileal tube with a suitable caliber. The latter was implanted into the bladder by submucosal (nonrefluxing) ileovesicostomy. RESULTS: Mean followup +/- SD was 9.6 +/- 2.4 months (range 6 to 13). Mean serum creatinine remained stable in all patients. Split kidney function (mercaptoacetyltriglycine clearance) was stable in 7 cases and improved in 3. Antegrade urography revealed dynamic unidirectional nonobstructed flow. Reflux was noted in only 1 case. Excretory urography and/or magnetic resonance urography showed excellent configuration of the substitute without evidence of dilatation or obstruction. CONCLUSIONS: The new technique offers certain distinct advantages. A short bowel segment is included with the consequent absence of metabolic complications. It allows construction of an ileal ureter with a suitable cross-sectional diameter without the need for tailoring and makes possible the use of an antireflux technique. PMID- 12576850 TI - Retroperitoneal bronchogenic cyst managed with retroperitoneoscopic surgery. PMID- 12576851 TI - Giant seminoma in a patient with 5 alpha-reductase type 2 deficiency. PMID- 12576853 TI - Successful treatment of dermatomyositis associated with adenocarcinoma of the prostate after radical prostatectomy. PMID- 12576852 TI - Lymphoma of the prostate and bladder presenting as acute urinary obstruction. PMID- 12576854 TI - Management of retained retractor during radical perineal prostatectomy. PMID- 12576855 TI - Vesicovaginal fistula after cervical cerclage: repair by transurethral suture cystorrhaphy. PMID- 12576856 TI - Priapism secondary to hypertriglyceridemia. PMID- 12576857 TI - Testosterone induced priapism in Kallmann's syndrome. PMID- 12576858 TI - Re: Extended pelvic lymphadenectomy in patients undergoing radical prostatectomy: high incidence of lymph node metastasis. A. Heidenreich, Z. Varga and R. von Knobloch. J. Urol, 167: 1681-6, 2002. PMID- 12576859 TI - Ectopic ureterocele: clinical application of classification based on renal unit jeopardy. AB - PURPOSE: The Churchill classification system grades the ectopic ureterocele based on the number of renal units in jeopardy. We apply this system to our experience to help predict response to initial surgical intervention. MATERIALS AND METHODS: All cases of ectopic ureterocele diagnosed at our institution between 1990 and 2000 were retrospectively reviewed and categorized using the Churchill classification system. Initial surgical procedure and subsequent outcomes were analyzed. RESULTS: A total of 53 patients were identified, of whom 42% had only upper pole involvement (grade I), 40% ipsilateral upper and lower pole involvement (grade II) and 19% bilateral renal involvement (grade III). Endoscopic incision was definitive in 56% of grade I, 20% of grade II and no grade III ureteroceles. An upper tract approach was definitive in 85% of grade I, 45% of grade II and no grade III ureteroceles. An initial lower tract procedure was successful in 80% of grade II and 100% of grade III ureteroceles. Overall the initial surgical procedure was definitive in 73% of grade I, 48% of grade II and 20% of grade III ureteroceles. CONCLUSIONS: Our results confirm the findings of Churchill. In this series the majority of grade I ureteroceles were successfully managed with an upper tract procedure. However, high grade ureteroceles were more likely to require a secondary operation unless an initial lower tract reconstruction was performed. PMID- 12576860 TI - Metanephric stromal tumor with urothelial extension. PMID- 12576861 TI - Vascularization of the hypospadiac prepuce and its impact on hypospadias repair. AB - PURPOSE: Only sparse data are available on the blood supply of the hypospadiac prepuce. We investigated the development and course of the preputial blood vessels, and applied the results of this study to surgery for hypospadias. MATERIALS AND METHODS: In 157 patients who underwent hypospadias surgery from October 1996 to December 2000 the prepuce was illuminated by a front and back lighting technique using endoscopic cold light and then photographed. Its blood vessel course and development were schematically presented. In 65 patients in whom the prepuce was not used for urethroplasty or penile body skin reconstruction the specimen was removed and its blood vessels were identified after injection with gelatin and Indian ink. The results of the 2 methods were compared. RESULTS: Based on the predominant blood vessels we classified preputial vascularization in hypospadias cases into groups, including 1-1 blood vessel predominant in 43%, 2-2 blood vessels predominant in 12%, 3-an H-like form with communication between 2 well developed blood vessels in 14% and 4-a net-like form with no predominant blood vessels in 29%. After comparing the results of the illumination technique versus specimen study in the same preputial tissue the illumination method was confirmed to be precise and easy to perform. CONCLUSIONS: Vascularization of the prepuce is crucial for hypospadias repair. Identification with high precision is achieved by the illumination technique. This method helps create the best vascularized flaps, whether longitudinal, oblique or spiral. PMID- 12576862 TI - Continent urinary diversion and the exstrophy-epispadias complex. AB - PURPOSE: The use of continent urinary reservoirs has gained wide acceptance, particularly in urinary reconstruction in children with a small capacity or neuropathic bladder. When augmentation cystoplasty is combined with clean intermittent catheterization, patients are often able to achieve continence with low intravesical filling pressures and renal preservation. Often this approach requires fashioning a continent cutaneous stoma, which remains the most challenging aspect of continent urinary reservoirs. We analyzed our experience with continent diversion in patients with exstrophy-epispadias to determine complications and long-term results. MATERIALS AND METHODS: We performed a retrospective database review of 704 cases of exstrophy-epispadias. Medical records were then used to identify those patients who had undergone creation of a continent urinary reservoir. Charts were reviewed to determine initial diagnosis, augmentation technique, continence mechanism, age, preoperative and postoperative bladder capacity, continence status and complications. RESULTS: Of the 91 patients identified (68 male, 23 female) who had undergone continent urinary diversion classic bladder exstrophy was present in 80, cloacal exstrophy in 8, complete male epispadias in 2 and female epispadias in 1. The most common techniques for augmentation and continent diversion were ileocystoplasty (41 patients [45%]) and sigmoid cystoplasty (30 [33%]), respectively. Appendix was used in 67 patients (74%) and variants of the Mitrofanoff procedure using segments of tapered ileum or ureter were used to create a continent stoma in 10 (11%). Bladder neck transection was performed in 59 patients (65%). Mean age at augmentation and continent diversion was 8 years (range 2 to 25), with a mean preoperative bladder capacity of 77 cc (15 to 220). Mean followup was 6 years (range 6 months to 12 years). Of the 91 patients 85 (93%) were continent with clean intermittent catheterization per stoma. Of these 85 patients 13 required anticholinergics and alpha-agonists to achieve continence. Six patients (7%) were incontinent after the procedure. Analysis of bladder capacity measurements after augmentation and continent diversion revealed that mean postoperative volume and mean volume increase were 404 cc (range 250 to 640) and 524%, respectively. The most common complications were bladder stone formation (24 patients [26%]) and stomal stenosis (21 [23%]). Bladder stones recurred in 9 patients and stomal stenosis in 3. Other less common complications were vesicourethral fistula (3 patients) and a small bladder perforation (2). CONCLUSION: Augmentation and continent diversion procedures can increase the functional capacity of the small contracted noncompliant exstrophic bladder, and allow the vast majority of patients to achieve continence and preserve renal function. Bladder calculi and stomal stenosis pose the most significant long-term complications in these patients. PMID- 12576863 TI - Treatment of phimosis with topical steroids in 194 children. AB - PURPOSE: Topical steroids have been advocated as an effective economical alternative to circumcision in boys with phimosis. We evaluated the effectiveness of topical steroid therapy as primary treatment in 194 patients with phimosis. METHODS: Between January 1996 and November 2000, 228 boys 16 years old or younger were referred for consideration of circumcision. When intervention was determined to be necessary, a 6-week course of topical steroids was used as primary treatment. Efficacy of treatment was evaluated at 3 months from initiation of therapy. RESULTS: Of the 228 patients 15 had such a mild degree of phimosis that no intervention was believed to be necessary, 19 were scheduled directly for circumcision due to cosmetic reasons, parent wishes, or severe phimosis with associated voiding problems and the remaining 194 received topical steroids as primary treatment. Of these 194 patients 25 had coexisting balanitis and 4 had a history of urinary tract infection. Conservative treatment was successful in 87%, 88% and 75% of patients with phimosis alone, coexisting balanitis and history of urinary tract infection, respectively. Overall, circumcision was avoided in 87% of patients treated with topical steroids. CONCLUSION: Topical steroids are becoming the standard conservative measure for treating phimosis. Our study supports this trend, with an overall efficacy of 87%. PMID- 12576864 TI - Endoscopic treatment with dextranomer-hyaluronic acid for vesicoureteral reflux: histological findings. AB - PURPOSE: Dextranomer-hyaluronic acid co-polymer is the first endoscopic bulking agent approved for vesicoureteral reflux in the United States. We evaluated the histopathological changes associated with this treatment in children with vesicoureteral reflux. MATERIALS AND METHODS: Children 1 to 11 years old in whom treatment with dextranomer-hyaluronic acid co-polymer for grades III or greater vesicoureteral reflux had failed were eligible for the study. Failure was defined as persistent vesicoureteral reflux on voiding cystourethrography done approximately 3 months after implantation. At ureteral reimplantation the implant and surrounding ureteral tissue were resected and fixed for histopathological analysis. Tissue sections (4 to 5 microm.) were stained for routine histology and examined under a light microscope. Patients with a similar grade of vesicoureteral reflux who had not undergone endoscopic treatment served as the control group. RESULTS: The study population comprised 23 patients with vesicoureteral reflux, of whom 13 with a mean age of 2 years 8 months at diagnosis underwent 1 to 3 treatments with dextranomer-hyaluronic acid co polymer. The remaining 10 patients with a mean age of 1 year 10 months at diagnosis did not receive the bulking agent before ureteral reimplantation. The implant remained in situ 13 to 39 months (mean 22). On ureteral reimplantation the implant was located at the site of injection in 12 of the 13 patients. Histologically a granulomatous inflammatory reaction indicated by giant cell infiltration was observed at the implantation site. At ureteral reimplantation 9 implants were pseudo-encapsulated. Calcification was present in 9 ureters, while the eosinophil count was greater than 5 cells per 0.125 mm2 in 7 ureters treated with dextranomer-hyaluronic acid co-polymer. Mast cell infiltration was similar in the treatment and control groups. CONCLUSIONS: Endoscopic treatment with dextranomer-hyaluronic acid co-polymer for vesicoureteral reflux is associated with a granulomatous reaction of the giant cell type, inflammatory cell infiltration and implant pseudo-encapsulation. They are typical histological findings associated with implantation of a foreign material. Dextranomer hyaluronic acid co-polymer remains safe and effective for vesicoureteral reflux in children. PMID- 12576865 TI - The outcome of bladder neck closure in children with severe urinary incontinence. AB - PURPOSE: We evaluated the outcome of children who underwent bladder neck closure for the management of severe urinary incontinence. MATERIALS AND METHODS: Bladder neck closure was performed in 12 patients with bladder exstrophy, 4 with myelomeningocele and 4 with urogenital sinus anomalies for severe urinary incontinence that persisted after multiple failed bladder outlet procedures. The effectiveness of bladder neck closure and the need for subsequent surgeries were ascertained. RESULTS: Mean followup was 5.4 years (range 1 to 12). At 3 months after surgery 40% of the patients were completely dry, 20% had leakage via the stoma and 40% had a urethral fistula. After additional surgeries 85% of the patients were completely continent 2 years following bladder neck closure (15% declined further surgeries or had an ileal conduit). Of the 15 patients with followup longer than 3 years only 40% remained completely dry, and leakage via the stoma developed in 47% after being dry for 1 year. Repeat urodynamic studies did not show any adverse changes in bladder dynamics in the incontinent patients. Stomal stenosis occurred in 30% of the patients and bladder stones developed in 40%. None of the patients had hydronephrosis or bladder perforation. The early complication of fistula formation and the late development of leakage via the stoma appear to be related in part to compliance with intermittent catheterization. CONCLUSIONS: Bladder neck closure is an effective method of achieving urinary continence in children in whom other bladder outlet surgery has failed. However, its success is dependent in part upon compliance with intermittent catheterization. Finally children who have undergone bladder neck closure are at an increased risk for stomal stenosis and bladder stones. PMID- 12576867 TI - Editorial: potential clinical use of exploiting metastasis suppressor genes to regulate prostatic cancer. PMID- 12576866 TI - Metastasis suppression: the evolving role of metastasis suppressor genes for regulating cancer cell growth at the secondary site. AB - PURPOSE: The prevention and treatment of prostate cancer metastasis continue to provide a significant clinical challenge. Identification of the rate limiting steps of metastasis and their underlying molecular mechanisms may lead to new therapeutic targets and also allow more accurate risk stratification for clinical metastases. We review the literature supporting growth of disseminated tumor cells at the secondary site as a key rate limiting step in metastasis. We also reviewed the definition, identification and characterization of metastasis suppressor genes, and discuss their evolving role in regulating this step. MATERIALS AND METHODS: We performed MEDLINE searches and manual bibliographic reviews on the specific steps of metastasis, including growth at the secondary site. In addition, we performed a comprehensive literature review to identify genes fitting the classic definition of a metastasis suppressor gene. The literature was also searched to assess the status of each gene in clinical cancer and evaluate functional support for the potential involvement of each gene in regulating growth at the secondary site. RESULTS: Clinical studies in prostate cancer and other cancer types suggest that dissemination to the secondary site is often an early clinical event. However, not all patients with tumor cells at the secondary site have overt metastatic lesions even in the absence of therapy, suggesting that growth at the secondary site may be highly inefficient. Complimentary approaches have allowed researchers to document and quantify the inefficiency of cancer cell growth at the secondary site. Regarding the mechanism of growth control, many studies support a role for the interaction of a cancer cell and the microenvironment at the secondary site influencing whether growth into metastasis may occur. The 7 genes that suppress metastasis without affecting primary tumor growth that have been identified are KAI1, CD44, mitogen activated protein kinase (MAPK) kinase 4, nm23-H1, nm23-H2, KiSS1 and BrMS1. Three of these genes (KAI1, CD44 and MAPK kinase 4) act as metastasis suppressor genes of prostate cancer, while the remainder have yet to be tested in this cancer type. Loss of expression has been demonstrated for most of these genes during the clinical progression of prostate cancer to metastasis. MAPK kinase 4 and KiSS1 appear to suppress metastasis by inhibiting cancer cell growth at the secondary site. Interestingly many metastasis suppressor genes have common roles in growth control, adhesion and cytoskeletal reorganization, suggesting a common mechanism of metastasis suppression. Proposed candidate pathways include signaling through Src kinase and Rac GTPase. CONCLUSIONS: The findings discussed support growth at the secondary site as a clinical target for metastasis treatment and prevention. Metastasis suppressor genes may offer valuable mechanistic insight for guiding specific therapeutic strategies, which may include drug induced reactivation of metastasis suppressor genes and their signaling pathways. Clinical assessment of metastasis suppressor gene product status in disseminated cancer cells may improve the accuracy of predicting the prognosis in patients with clinically localized disease. PMID- 12576868 TI - Connexin 26 is abnormally expressed in bladder cancer. AB - PURPOSE: Connexin 26 is the major gap junction protein in urothelial and mammary epithelial cells, and a putative tumor suppressor gene. We evaluated connexin 26 expression in normal urothelium and in bladder cancer. MATERIALS AND METHODS: A total of 40 formalin fixed, paraffin embedded bladder tumors and 5 normal urothelial specimens were analyzed by immunohistochemistry. Two observers visually scored connexin 26 expression in these specimens. RESULTS: Normal urothelium expressed connexin 26 in a punctate staining pattern with limited expression in the basal layer. Decreased connexin 26 expression was observed in 28 of 40 tumors (70%). Connexin 26 was diffusely expressed in 5 of 18 low grade, noninvasive tumors (28%), whereas loss of expression was observed in heterogeneous (30% to 70% positive staining) or extensive (less than 30% positive staining) fashion in 8 (44%) and 5 (28%), respectively. Seven of 22 high grade or invasive tumors (32%) showed diffuse connexin 26 expression, whereas expression was decreased in a heterogeneous or extensive pattern in 9 (41%) and 6 (27%), respectively. Intracytoplasmic localization of connexin 26 was also observed. CONCLUSIONS: Expression of connexin 26 is altered in bladder cancer. These aberrant patterns of connexin 26 expression may contribute to the malignant phenotype of this disease. PMID- 12576869 TI - Quantitative GSTP1 methylation clearly distinguishes benign prostatic tissue and limited prostate adenocarcinoma. AB - PURPOSE: Hypermethylation of the glutathione S-transferase gene (GSTP1) is the most common (greater than 90%) reported epigenetic alteration in prostate cancer. It occurs early in cancer progression and it is a promising marker for detecting organ confined disease. To evaluate its use as a diagnostic tool for cancer we used quantitative GSTP1 methylation to test for the presence of cancer in 45 prostate needle biopsy samples. MATERIALS AND METHODS: Paraffin tissue samples from 45 patients with minute foci of intermediate grade prostatic adenocarcinoma or benign disease on needle biopsy were tested for GSTP1 hypermethylation using quantitative fluorogenic real-time methylation specific polymerase chain reaction. This assay was performed in blinded fashion without previous knowledge of the histopathological diagnosis. RESULTS: DNA from 29 of the 45 paraffin samples was amenable to polymerase chain reaction amplification. In these 29 samples GSTP1 methylation was detected in 11 of 15 cases of limited cancer and in 0 of 14 of benign disease (2-sided Fisher's exact test, p <0.0001). Thus, this assay had 73% sensitivity, 100% specificity, 100% positive and 78% negative predictive values. CONCLUSIONS: Quantitation of GSTP1 hypermethylation accurately detects the presence of cancer even in small, limited tissue samples. It represents a promising diagnostic marker that could be used as an adjunct to tissue biopsy as part of prostate cancer screening. PMID- 12576870 TI - Suppression of prostate cancer induced bone remodeling by the endothelin receptor A antagonist atrasentan. AB - PURPOSE: We examined the effects of atrasentan (endothelin-A receptor antagonist) on bone deposition and resorption markers and on bone scan index. MATERIALS AND METHODS: This double-blind, randomized, placebo controlled clinical trial of hormone refractory prostate cancer patients was done at 74 medical centers in the United States and Europe. A total of 288 asymptomatic patients with hormone refractory prostate adenocarcinoma and evidence of metastatic disease were randomized to 1 of 3 treatment groups, namely 2.5 mg. atrasentan, 10 mg. atrasentan or placebo administered orally daily until disease progression. The main outcomes measures were changes in bone deposition markers (total alkaline phosphatase and bone alkaline phosphatase) and bone resorption (N-telopeptides, C telopeptides and deoxypyridinoline), and in the bone scan index. RESULTS: At baseline markers of bone deposition and resorption were elevated 1.4 to 2.7-fold above respective upper limits of normal. Subjects receiving placebo experienced a 58% elevation in mean total alkaline phosphatase and a 99% elevation in mean bone alkaline phosphatase (p < 0.001), whereas subjects receiving 10 mg. atrasentan maintained stable mean total alkaline phosphatase and bone alkaline phosphatase values compared with baseline. N-telopeptides, C-telopeptides and deoxypyridinoline elevation from baseline were consistently less in patients receiving 10 mg. atrasentan compared with placebo. Similar trends were observed in subjects who received 2.5 mg. atrasentan. Changes in clinical bone scan studies paralleled bone marker changes. CONCLUSIONS: Atrasentan suppressed markers of biochemical and clinical prostate cancer progression in bone and demonstrates clinical activity for hormone refractory prostate cancer. PMID- 12576871 TI - Anoikis induction by quinazoline based alpha 1-adrenoceptor antagonists in prostate cancer cells: antagonistic effect of bcl-2. AB - PURPOSE: The ability of the quinazoline derived alpha1-adrenoceptor antagonists doxazosin and terazosin to induce apoptosis in benign and malignant prostate cells has been established. In this study we investigated the effect of the 2 piperazidinyl quinazoline based alpha1-adrenoceptor antagonists and the methoxybenzene sulfonamide alpha1-antagonist tamsulosin on human prostate cancer cell adhesion. MATERIALS AND METHODS: Androgen independent PC-3 prostate cancer cells and PC-3 transfectant clones over expressing the apoptosis suppressor bcl-2 were used as an in vitro model. Cells were treated with pharmacologically relevant doses of 1 of the 3 alpha1-adrenoceptor antagonists and the effect on cell viability/cell adhesion on various substrates was examined. Analysis of expression of key attachment factors, such as vascular endothelial growth factor (VEGF) and hypoxia inducible factor-alpha, was performed. RESULTS: Our results indicate a significant decrease in prostate cancer cell adhesion to gelatin and collagen but not to fibronectin in prostate cancer cells treated with doxazosin or terazosin (25 microM.) compared with untreated control cultures (p <0.05). In contrast, tamsulosin had no effect on prostate cancer cell adhesion. The 2 quinazolines doxazosin and terazosin but not tamsulosin had a significant inhibitory effect on prostate tumor cell invasion. In bcl-2 over expressing prostate cancer cells there was significant suppression of doxazosin induced anoikis and cell invasion compared with neocontrol transfectants (p <0.05). Doxazosin resulted in transient down-regulation (2-fold decrease) of VEGF at the mRNA and protein levels, as detected by reverse transcriptase-polymerase chain reaction and Western blotting, respectively. No significant changes in the expression profile of hypoxia inducible factor-1 alpha were observed after treatment with quinazolines. Furthermore, bcl-2 resulted in partial reversion of the doxazosin induced VEGF decrease. CONCLUSIONS: These findings demonstrate that the quinazoline derived alpha1-antagonists doxazosin and terazosin but not sulfonamide based tamsulosin induce anoikis and inhibit prostate cancer cell invasion, an effect that is antagonized by bcl-2. This molecular basis of an alpha1-adrenoceptor independent action against prostate cancer cells by the quinazolines may have potential therapeutic significance in prostate cancer. PMID- 12576872 TI - Maspin functions as tumor suppressor by increasing cell adhesion to extracellular matrix in prostate tumor cells. AB - PURPOSE: Maspin, a unique member of the serine protease inhibitor family, shows tumor suppressing activity for breast cancer progression and metastasis. Few studies have directly linked maspin function to prostate cancer. We used prostate tumor cells derived from the TRAMP (transgenic adenocarcinoma of mouse prostate) prostate tumor model to study the tumor suppressive function of maspin in prostate cancer. MATERIALS AND METHODS: Maspin cDNA was introduced via a retroviral plasmid into TRAMP C2N prostate tumor cells, which are aggressive and invasive in nature. We investigated the tumorigenesis of these stable cell lines in vitro by assessing the growth rate, anchorage independence and adhesion to extracellular matrix proteins such as fibronectin and laminin. RESULTS: Stable cell lines expressing maspin had decreased tumorigenic potential, as assessed by anchorage independent growth in soft agar assay compared with controls. Maspin stable transfectants showed decreased metastatic potential, as evaluated by modified Boyden chamber assay and increased adhesion to fibronectin and laminin. CONCLUSIONS: Our study confirms that maspin has a tumor suppressive role not only in breast cancer, but also in prostate cancer. The data in this study suggest that maspin can decrease the tumorigenic and metastatic potential of prostate tumors, most probably by remodeling cell-extracellular matrix interactions or triggering extracellular matrix mediated signaling pathways that negatively regulate tumor migration and invasion. PMID- 12576873 TI - Editorial: maspin as a molecular target for cancer therapy. PMID- 12576875 TI - Expansion and fixation properties of a new braided biodegradable urethral stent: an experimental study in the rabbit. AB - PURPOSE: Biodegradable spiral urethral stents have been used with favorable results combined with thermal treatments of the prostate and for recurrent urethral strictures but the configuration of the helical spiral is not ideal. We developed a new tubular mesh configuration for the biodegradable urethral stent and evaluated its expansion and locking properties in the rabbit urethra. MATERIALS AND METHODS: The stents were made of self-reinforced polylactic acid polymer (Bionx Implants, Ltd., Tampere, Finland) blended with BaSO (Alfa Chem, New York, New York) to achieve radiopacity. Two braiding patterns, that is 1 over 1 and 2 over 2 + 1, were used to produce a tubular mesh structure. Stainless steel stents (pattern 1 over 1) served as controls. The stents were inserted into the posterior urethra of 27 male rabbits. The animals were sacrificed after 1 week, 1 and 6 months, respectively. X-rays were done immediately after stent insertion and at sacrifice. Longitudinal movement and expansion were assessed on the x-rays.(4) RESULTS: All stents maintained position in the urethra without fixation. Macroscopic disorientation of the structure of the 2 over 2 + 1 braided self-reinforced polylactic acid polymer stents began before 1 month, while 1 over 1 braided stents retained their construction. At 6 months 3 of 6 biodegradable stents were degraded. Average longitudinal movement was 2 mm. (range 1 to 3) in the 1 over 1 self-reinforced polylactic acid polymer group, 2 mm. (range 0 to 7) in the 2 over 2 + 1 polylactic acid group and 3 mm. (range 3 to 3) in controls at 1 month. CONCLUSIONS: Biodegradable polymers are suitable materials for braided urethral stents. The expansion properties of the 2 braiding models tested in this study sufficed to fix the stents in situ in the prostatic urethra. However, the 1 over 1 braiding pattern was superior to the 2 over 2 + 1 pattern, in that it retained its macroscopic construction until the degradation of single self reinforced polylactic acid polymer fibers. PMID- 12576874 TI - Platelet derived growth factor-BB is a potent mitogen for rat ureteral and human bladder smooth muscle cells: dependence on lipid rafts for cell signaling. AB - PURPOSE: Fibromuscular tissues of the detrusor/bladder body (B), trigone (T) and ureter (U) display distinct patterns of tissue remodeling in pathologic contexts, however the mechanisms underlying these observations are unknown. In this study we asked whether B, T and U smooth muscle cells (SMC) respond to several SMC growth factors and explored the role of caveolae/lipid raft membrane microdomains in signaling by one of these factors, PDGF-BB. MATERIALS AND METHODS: SMC were isolated and cultured from B, T and U from newborn rats and from human bladder detrusor. Responses to growth factors were assessed by cell proliferation, DNA synthesis, and immunoblot methods. Cholesterol was depleted from cell membranes in select experiments using cyclodextrin and the cholesterol synthesis inhibitor lovastatin. High-affinity PDGF receptor (PDGFR) sites were measured by 125I-PDGF BB binding assay. RESULTS: PDGF-BB increased DNA synthesis rate in U and T SMC, with U SMC being highly responsive; in contrast, B SMC did not respond to this growth factor. Two other mitogens, HB-EGF and FGF-2, marginally stimulated DNA synthesis in all lineages. Human detrusor (hD) SMC were also highly responsive to PDGF-BB. Differences in responses to PDGF-BB correlated with translocation of PDGFRs into the caveolae/lipid raft membrane fraction following stimulation, but not with the number of high affinity PDGF binding sites. Cholesterol depletion from cell membranes reduced the response of U and hD SMC to PDGF-BB. CONCLUSIONS: These findings indicate that 1) PDGF-BB is likely to be a physiologically relevant stimulator of mitogenic signaling in certain types of urinary tract SMC, 2) there are significant and unanticipated regional differences in the ability of urinary tract SMC to respond to muscle mitogens, and 3) lipid raft membrane microdomains mediate, in part, the ability of urinary tract SMC to respond to PDGF-mediated signals. PMID- 12576876 TI - Penile weight and cell subtype specific changes in a post-radical prostatectomy model of erectile dysfunction. AB - PURPOSE: We evaluated neurogenic erectile dysfunction, focusing on the post radical prostatectomy model. We investigated changes in DNA, protein and apoptotic cells of the rat penis after denervation. Gross morphometry was measured to elucidate the impact of chemical changes. MATERIALS AND METHODS: Postpubertal male Sprague-Dawley rats were randomized to bilateral or unilateral cavernous nerve transection, or sham operation. Wet weight, DNA content and protein content were measured. Tissue sections were stained for apoptosis by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling and the apoptotic index was calculated. Dual staining was performed for endothelial and smooth muscle cells to identify apoptotic cells. RESULTS: Penile wet weight was significantly decreased at all time points after bilateral neurotomy (p <0.0005). Unilateral neurotomy allowed much greater preservation of penile weight. DNA content was significantly decreased in bilaterally denervated penes and unchanged in unilaterally operated penes. Protein content was not significantly altered in the bilateral or unilateral cohorts. Bilateral neurotomy induced significant apoptosis, while unilateral surgery caused significantly less apoptosis. Each population had apoptotic clustering just beneath the tunica albuginea, which was mostly smooth muscle cells. CONCLUSIONS: These data suggest the importance of neural integrity to maintain penile homeostasis. The loss in penile weight was consistent with the anecdotal experience of many clinicians. Decreased DNA content may have been due to significant levels of apoptosis in smooth muscle cells. Preserved protein content may suggest an increase in extracellular protein, as postulated in corporeal fibrosis. The subtunical population of apoptotic smooth muscle cells revealed a mechanism for veno occlusive dysfunction observed after radical prostatectomy. These effects were significantly moderated in the unilateral model, reinforcing the critical nature of neural integrity. PMID- 12576877 TI - Amphetamine withdrawal does not produce a depressive-like state in rats as measured by three behavioral tests. AB - Administration of amphetamine (AMPH) can induce symptoms of psychosis in humans and locomotor sensitization in rats; in contrast, withdrawal from a period of AMPH intake is most often associated with symptoms of human endogenous depression. The aim of this study was to determine whether AMPH withdrawal produces a depressive-like state in rats. The present study examined the effects of withdrawal from an escalating-dose AMPH schedule (ESC; three daily injections over 6 days, 1-5 mg/kg, i.p.) and an intermittent-dose AMPH schedule (INT; one daily injection over 6 days, 1.5 mg/kg, i.p.) on animals' performance in three behavioral paradigms related to depression: the Porsolt swim test, the learned helplessness assay and operant responding for sucrose on a progressive ratio schedule. ESC and INT AMPH withdrawal had no effect on any of these tests or on stress responsiveness as measured by increased plasma levels of corticosterone (CORT) and adrenocorticotropin following the swim test, although basal CORT levels were higher in AMPH-withdrawn animals compared to controls. Finally, we confirmed the presence of locomotor sensitization for both AMPH schedules after 30 days of withdrawal. Our results suggest that the ability of AMPH withdrawal to produce symptoms of depression may not be evident in all behavioral screens for depressive symptoms in the rat. PMID- 12576878 TI - The prediction of individual differences in response to D-amphetamine in healthy adults. AB - Based on previous animal work, the present study investigated whether individual differences in motor activity in a novel environment predicted reinforcing and behaviorally activating effects of D-amphetamine (0, 5, 10 and 20 mg p.o.) in healthy adults. When exposed to a novel environment, 18 participants had high levels of motor activity (high responders; HR) and six had low levels (low responders; LR). These group differences were used to predict effects of D amphetamine on drug reinforcement, salivary cortisol, motor activity, subjective effects, and acoustic startle reflex in subsequent sessions. Unlike observations in rodents, (1). dose-dependent reinforcing effects of D-amphetamine were evident but without group differences; (2). motor activity was greater in HR but did not vary with D-amphetamine dose; and (3). cortisol levels were not related to the reinforcing effects of D-amphetamine. Startle reflex amplitudes were greater in HR following placebo, but D-amphetamine 20 mg equalized this group difference. There was a trend towards less prepulse inhibition of the acoustic startle reflex in HR compared to LR. LR reported greater overall negative effect following amphetamine administration, but this was not consistently related to dose. Finally, participants with high sensation-seeking personality scores exhibited less prepulse inhibition. The results are discussed in terms of the extant literature. PMID- 12576879 TI - The effects of acutely administered cocaine on responding maintained by a progressive-ratio schedule of food presentation. AB - Lever pressing in rats (N=5) was reinforced under a progressive-ratio (PR) schedule of food presentation, in which the number of responses required increased exponentially. The session was terminated when 1 h passed without completion of the scheduled ratio. Doses of cocaine (5.6-42.0 mg/kg; one subject received a dose of 56.0 mg/kg) as well as saline were administered i.p. prior to the session. Under non-drug conditions, breakpoints were typically less than 100, and substantial responding usually occurred only during about the first 10 min of the session. The rate of responding usually increased over the first 2-8 reinforcers and then decreased for the last few reinforcers obtained. For four of five rats, breakpoint, overall rate of response, and session duration were first increased above control and vehicle levels by increasing doses of cocaine. Larger doses produced smaller increases, no effect, or decreases. Cocaine, in the range of doses near the apex of the breakpoint dose-effect functions, suppressed rates of responding at the small ratios present at the beginning of the session. It is suggested that cocaine increases low rates of response if: (1). rates are low due to extinction; and (2). the stimuli present are those present when the response is reinforced. PMID- 12576880 TI - Morphine state-dependent learning: interactions with alpha2-adrenoceptors and acute stress. AB - The interactions of -adrenoceptors and acute restraint stress with morphine state dependent memory of passive avoidance were examined in mice. Memory acquired following pre-training morphine administration (5 mg/kg, i.p.) was dose- and time dependently retrieved by pre-test morphine; this effect was reversible by yohimbine (1 mg/kg). Pre-test clonidine (0.005-0.1 mg/kg) was also effective in restoring morphine-induced memory. Pre-training clonidine (2 mg/kg) induced an amnestic effect that was restorable by pre-test clonidine or morphine; this effect was also blocked by yohimbine. Acute pre-training stress for 2 h induced an amnestic effect that was reversible by pre-test morphine (1 and 5 mg/kg) or clonidine (0.01 and 0.1 mg/kg). Finally, acute pre-test stress could restore the impairment of memory induced by pre-training morphine. The data are suggestive of a functional interaction between -opioid, -adrenergic receptors and stress in modulating state-dependent learning and memory. PMID- 12576881 TI - Catalepsy intensifies context-dependently irrespective of whether it is induced by intermittent or chronic dopamine deficiency. AB - It is well known that neuroleptic-induced catalepsy in rats intensifies upon repeated testing. Here, the question is addressed whether intensification of catalepsy results from intermittent drug administration or from intermittent context exposure. In experiment 1, rats were treated with intermittent haloperidol injections (0.25 mg/kg) followed by the catalepsy test (descent latency from the horizontal bar). In experiment 2, rats were lesioned with 6 hydroxydopamine injections into the striatum, resulting in a 45% reduction of dopamine concentration. Catalepsy was tested intermittently for several weeks. In both experiments we found a very stable intensification of catalepsy over 9 (haloperidol rats) and 11 (lesioned rats) days, showing that intensification is not due to intermittent dopamine depletion. In both experiments, intensification of catalepsy was very stable and was observed 18 days later in haloperidol treated rats and 101 days later in lesioned animals. However, a change of the environmental context abolished the intensified catalepsy in both experiments. It is concluded that intensification of catalepsy is due to intermittent context exposure rather than intermittent drug administration. It is generally accepted that 6-hydroxydopamine lesions represent an animal model of Parkinson's disease. Given the results above, context-dependent intensification of parkinsonian symptoms might also occur in Parkinson's disease, and its prevention should be taken into consideration for future therapy of the disease. PMID- 12576882 TI - Nicotine trace discrimination in rats with midazolam as a mediating stimulus. AB - It was shown previously that effects of drugs present prior to training sessions could serve as discriminative stimuli. Further experiments have aimed to determine whether a second drug can serve as a mediating stimulus that increases the strength of stimulus control by such pre-session drug effects. Rats were trained in a two-lever discrimination procedure with food reinforcers presented on a tandem variable-interval fixed-ratio (VI-FR) schedule. Injections of nicotine (0.6 mg/kg) or saline were followed after 5 min by administration of midazolam (0.2 mg/kg) as a putative mediating stimulus. The nicotine antagonist mecamylamine (1.0 mg/kg) was administered 5 min after midazolam, to block effects of nicotine during training sessions, as in previous work on pre-session drug effects. Stimulus control was acquired slowly and to an accuracy of only 75%. Midazolam did not facilitate the acquisition or magnitude of nicotine-induced stimulus control. However, extinction tests showed that the presence of midazolam was required for expression of stimulus control by pre-session effects of the training dose of nicotine. The response to nicotine (0.075-0.6 mg/kg) was dose related, but the dose-response relationship was not dependent upon the presence of midazolam. In a group of rats trained with nicotine and midazolam as above, but without mecamylamine, stimulus control by nicotine was not dependent upon the presence of midazolam. In all cases, overall rates of responding were very low when tests were carried out without midazolam, suggesting the presence of state dependent learning. The results imply that under appropriate conditions the discriminative stimulus effects of one drug (nicotine) can be mediated by the action of a second substance (midazolam). This finding can be conceptualized in terms of occasion setting, with nicotine serving as the feature and midazolam as the target stimulus. Furthermore, it appears that even when rates of responding show drug-state dependence, this is not necessarily the case for discriminative stimulus effects. PMID- 12576883 TI - Effects of positive GABA(A) modulators on a multiple-component, repeated acquisition test of spatial learning. AB - The purpose of this study was to determine the effects of the benzodiazepines, midazolam and chlordiazepoxide, and the barbiturate, pentobarbital, on spatial learning, in a within-subject, repeated-acquisition and performance procedure adapted to the Morris Swim Task. In the presence of one stimulus arrangement, rats learned to swim to a hidden escape platform that was always in the same location in a swimming pool (performance component). In the presence of a second stimulus arrangement, the platform moved to a different place in the pool for each daily session (acquisition component). All subjects completed six training trials in both components during each daily training session, alternating between the two components within each session. Relatively direct paths to the platform and short escape latencies in the performance component, and steep within-session learning curves in the acquisition component, demonstrated that behavior under each component was controlled by the discriminative stimuli. All three GABA(A) modulators increased swim distances, escape latencies, and slowed swim speed in a dose-dependent manner. Midazolam and chlordiazepoxide, but not pentobarbital, produced selective impairments of swim distances and escape latencies in the acquisition component. Benzodiazepines disrupted acquisition at doses that did not disrupt steady-state performance. Pentobarbital impaired acquisition only at doses that also disrupted behavior during the performance component and reduced swimming speeds. PMID- 12576884 TI - Sex differences in (-)-pentazocine antinociception: comparison to morphine and spiradoline in four rat strains using a thermal nociceptive assay. AB - The present study examined the influence of sex on the antinociceptive effects of (-)-pentazocine, morphine and spiradoline in four rat strains, using a warm-water (50, 52 and 55 degrees C) tail-withdrawal procedure. In F344, Lewis, Sprague Dawley (SD) and Wistar rats, baseline latencies decreased with increases in water temperature, and at each water temperature latencies were longer in males than in their female counterparts. Morphine and spiradoline produced maximal or near maximal antinociceptive effects in males and females of each strain. Whereas morphine was generally more potent in males, sex differences were not consistently observed with spiradoline. In contrast, there were marked sex differences with (-)-pentazocine, and in each strain (-)-pentazocine was more potent and produced a greater maximal effect in males. The magnitude of the sex differences varied markedly across strains, with (-)-pentazocine being 2.5-fold more potent in males of the F344 strain, but 11-fold more potent in males of the Wistar strain. When collapsed across nociceptive stimulus intensities, sex differences were largest in the Wistar and Lewis strains and smallest in the SD and F344 strains. The present findings indicate that there are marked sex differences in (-)-pentazocine antinociception, and that the magnitude of this effect is genotype dependent. PMID- 12576885 TI - The antidepressant-like effects of neurokinin NK1 receptor antagonists in a gerbil tail suspension test. AB - Recent clinical evidence supports the potential of neurokinin NK1 receptor antagonists as novel antidepressant drugs. A number of NK1 antagonists have reduced affinity for rat and mouse NK1 receptors compared to human, making it difficult to test for efficacy in traditional animal models. NK1 antagonists, in general, have similar affinity at gerbil and human NK1 receptors. The aims of these studies were first, to validate the gerbil tail suspension test, a test used frequently to demonstrate antidepressant drug efficacy in mice, and second, to determine whether the test could be used to demonstrate the antidepressant potential of NK1 antagonists. Immobility time was reduced by oral administration of the antidepressants imipramine (3-30 mg/kg), desipramine (1-30 mg/kg), amitriptyline (30 mg/kg), fluoxetine (1-30 mg/kg), paroxetine (3-10 mg/kg), citalopram (0.1-3 mg/kg), sertraline (1-30 mg/kg), venlafaxine (1-30 mg/kg) and nefazodone (100 mg/kg). Furthermore, oral administration of the NK1 antagonists MK-869 (10 mg/kg), L-742694 (10 mg/kg), L-733060 (10 mg/kg), CP-99994 (30 mg/kg), and CP-122721 (3-30 mg/kg) reduced immobility time. Diazepam (1-10 mg/kg), chlordiazepoxide (1-10 mg/kg), buspirone (3-30 mg/kg), FG-7142 (1-30 mg/kg), and haloperidol (1-10 mg/kg) did not reduce immobility. Amphetamine (0.3-10 mg/kg) and atropine (0.3-10 mg/kg) reduced immobility, suggesting susceptibility to false positives, e.g. compounds that affect locomotion. Compounds were therefore tested in a gerbil locomotor activity (LMA) test to ensure that the antidepressant-like effects were not secondary to effects on activity. Antidepressant drugs and NK1 antagonists had no effect on LMA at doses that reduced immobility, whereas amphetamine and atropine induced marked hyperactivity. These studies support both the utility of gerbils in behavioral pharmacology and the antidepressant potential of selective NK1 antagonists. PMID- 12576886 TI - Systemic and peritoneal inflammatory response after laparoscopic or conventional colon resection in cancer patients: a prospective, randomized trial. AB - PURPOSE: This study was designed to evaluate differences in both the peritoneal and systemic immune response after laparoscopic and conventional surgical approaches. METHODS: Patients with a primary carcinoma were prospectively randomized to curative laparoscopic (n = 12) or conventional (n = 14) colon resection. The proinflammatory cytokines interleukin-6, interleukin-8, and tumor necrosis factor-alpha were measured in the peritoneal drain fluid and in the serum. C-reactive protein and leukocyte counts and the differences in leukocyte subpopulations and expression of human leukocyte antigen-DR on monocytes were measured perioperatively. RESULTS: Significantly higher levels of proinflammatory cytokine were found in the peritoneal drain fluid than in the circulation after both procedures. Serum interleukin-6 and interleukin-8 levels were significantly lower 2 hours after laparoscopic surgery than with the conventional procedure. Postoperative cellular immune counts and human leukocyte antigen-DR expression normalized earlier after the laparoscopic approach. CONCLUSIONS: The systemic proinflammatory concentrations after both surgical approaches represent only a small fragment of what is generated in the peritoneal drain fluid. Even if the immediate levels of proinflammatory cytokines in the serum are significantly lower in the laparoscopic group, the same cytokines locally produced showed no differences, which suggests that the two intra-abdominal approaches are equally traumatic. No differences in cellular response were observed between the groups. PMID- 12576887 TI - Is routine pouch surveillance for dysplasia indicated for ileoanal pouches? AB - PURPOSE: Isolated accounts of neoplastic pouch transformation suggest the need to perform routine ileoanal pouch surveillance with biopsy. These reports imply a model of dysplasia to cancer transformation of the pouch mucosa. More recent reports studying "high risk" ulcerative colitis patients concluded that the development of pouch dysplasia is indeed a rare event. This study was designed to evaluate our institutional incidence of dysplasia in ileoanal pouch during long term follow-up. METHODS: A prospective database of all patients undergoing ileoanal pouch construction (n = 767) was queried for all patients undergoing pouch biopsy between 1983 and 2001. All patients with ulcerative colitis who underwent pouch biopsy were included. This excluded patients with Crohn's disease, indeterminate colitis, and familial adenomatous polyposis. Pathology reports were reviewed for histologic evidence of inflammation, atypia, metaplasia, dysplasia, or cancer. Patient age at biopsy, pouch age at time of biopsy, and pathology were analyzed. RESULTS: The ileoanal pouches of 160 patients were surveyed with biopsies a total of 222 times. The average length of follow-up from pouch construction to time of surveillance and biopsy was 8.4 +/- 4.6 years. There were 83 patients (52 percent) whose pouches were older than 10 years (mean, 12.7 +/- 2) at time of surveillance. With over 1,800 pouch-years of surveillance, only 1 patient had focal, low-grade dysplasia in the pouch. This patient demonstrated no evidence of dysplasia on further surveillance. CONCLUSION: Even with long-term follow-up of ileoanal pouch patients, there is little evidence to support routine biopsy of the ileal mucosa in ulcerative colitis patients. PMID- 12576888 TI - Survival after curative resection for mucinous adenocarcinoma of the colorectum. AB - PURPOSE: Previous reports have suggested that mucinous colorectal adenocarcinomas are more advanced at diagnosis and have a poorer prognosis than nonmucinous colorectal adenocarcinomas. The purpose of this study was to clarify whether the mucin-producing histologic type of carcinoma is associated with a worse prognosis than nonmucinous, differentiated colorectal adenocarcinoma for patients who undergo curative surgery. METHODS: Using a database of 2,678 surgical patients with colorectal cancers operated on at Aichi Cancer Center between 1965 and 1994, we investigated 97 cases of mucinous adenocarcinoma and 2,197 cases of nonmucinous adenocarcinoma. We also evaluated the outcomes of patients who underwent surgery with curative intent. To determine whether the mucinous adenocarcinoma itself was an independent prognostic factor in the curative resected patients, a multivariate analysis was performed. RESULTS: The mucinous adenocarcinoma patients were found to be younger (P = 0.0003), have more lymph node involvement (48.5 vs 40.3 percent; P = 0.0564), more peritoneal dissemination (19.6 vs 5.6 percent; P < 0.0001), greater frequency of advanced stage disease (P = 0.0006), a lower rate of curative resection (76.3 vs 84.4 percent; P = 0.0450), and lower overall 5-year survival rates (41 vs 62.4 percent; P = 0.0002) than nonmucinous adenocarcinoma patients. In the subjects who underwent curative resection, the 5-year survival rate for those with mucinous adenocarcinoma was significantly worse than for those with nonmucinous adenocarcinoma (54 vs 73.3 percent; P = 0.0020). Multivariate analysis using the Cox proportional hazards model showed that the clinically significant predictive factors were stage at diagnosis, mucinous histology, tumor location, gender and age. The mucinous histologic type itself was an independent factor for poor prognosis for patients who underwent curative surgery. CONCLUSIONS: In patients with colorectal carcinomas who underwent surgery with curative intent and who had colorectal carcinomas of the mucinous histologic type, there was significant correlation with prognosis as measured by overall survival rate after adjustment had been made for major confounders. PMID- 12576889 TI - Recovery of the rectoanal inhibitory reflex after restorative proctocolectomy: does it correlate with nocturnal continence? AB - PURPOSE: The rectoanal inhibitory reflex has an important role in fecal sampling and discrimination of rectal contents. The aim of this study was to determine the significance of rectoanal inhibitory reflex after restorative proctocolectomy with ileal pouch-anal anastomosis for mucosal ulcerative colitis. METHODS: The medical records of 345 patients who underwent ileal pouch-anal anastomosis from September 1988 to May 1999 were retrospectively reviewed. One hundred patients who underwent double-stapled ileal pouch-anal anastomosis and had anorectal physiology testing within 3 months before surgery as well as after ileostomy closure (mean, 23.1; range, 3-77 months) were analyzed. Anorectal physiology testing included detecting the presence of the rectoanal inhibitory reflex, sensory threshold volume, and rectal or pouch capacity and compliance. Parameters to determine incontinence included daytime and nocturnal bowel movement frequency, nocturnal spotting, status of continence for solid or liquid stool, gas, use of pads, and lifestyle alteration were surveyed in 62 of the 100 patients at a mean of 3.9 (range, 1-9.1) years to determine the incontinence score. RESULTS: Whereas the rectoanal inhibitory reflex was noted in 96 (96 percent) patients before surgery, it was found in only 53 (53 percent) after ileostomy closure (P < 0.0001). Incontinence status data was available in only 62 of the 100 patients (32 RAIR-positive; 30 RAIR-negative). There were no significant differences between the rectoanal inhibitory reflex-positive and the rectoanal inhibitory reflex-negative groups relative to the interval between surgery and manometry (22 vs 25 months), postoperative threshold sensation volume (32 vs 31 ml), postoperative compliance (19 vs 12 cm H(2)O/ml), postoperative capacity (85 66 ml), daytime/nighttime stool frequency (6.2/2 vs 5.5/1.5), or postoperative incontinence score (3.9 vs 1.8). However, there were significant differences relative to the incidence of nocturnal soiling (12/30 (40 percent) 23/32 (72 percent), P = 0.0012) favoring the presence of the rectoanal inhibitory reflex. CONCLUSION: Preservation of the rectoanal inhibitory reflex correlated with a decrease in the incidence of nocturnal soiling after double-stapled ileoanal reservoir construction. PMID- 12576890 TI - Surgical management of ulcerative colitis in the presence of primary sclerosing cholangitis. AB - INTRODUCTION: The surgical management of ulcerative colitis in the patient with primary sclerosing cholangitis is controversial. METHODS: This study was designed as a retrospective chart review of all patients with primary sclerosing cholangitis who were surgically treated for ulcerative colitis. RESULTS: Sixteen patients with primary sclerosing cholangitis and ulcerative colitis were identified. The indication for ulcerative colitis surgery was dysplasia in 7 patients (44 percent), cancer in 2 (13 percent), intractability in 4 (25 percent), and unknown in 1. Final colon pathology demonstrated cancer in three patients and dysplasia in four. Two patients had biliary cancer discovered at the time of orthotopic liver transplantation. Thirteen patients were known to have primary sclerosing cholangitis when they underwent surgery for ulcerative colitis; two patients with severe primary sclerosing cholangitis underwent simultaneous orthotopic liver transplantation/total abdominal colectomy and did well with subsequent ileal pouch reconstruction. Two patients had orthotopic liver transplantation first and then ileal pouch-anal anastomosis (1 patient) or total abdominal colectomy (1 patient) and did well. Seven patients had well controlled primary sclerosing cholangitis on medication and underwent ileal pouch anal anastomosis or total abdominal proctocolectomy without significant hepatic compromise. One patient with moderate primary sclerosing cholangitis underwent ileorectal anastomosis and had severe liver failure postoperatively but survived. Another patient with worsening primary sclerosing cholangitis after total abdominal colectomy has since developed persistent bleeding from peristomal varices. CONCLUSIONS: The overall cancer/premalignant lesion rate was high (50 percent in this study) in patients with primary sclerosing cholangitis and ulcerative colitis. Complications associated with the surgical management of ulcerative colitis are largely dictated by the degree of liver disease present at the time of surgery. Patients with significant primary sclerosing cholangitis that requires colectomy can undergo simultaneous orthotopic liver transplantation/total abdominal colectomy and then be candidates for subsequent ileal pouch-anal anastomosis reconstruction once liver function has improved. Patients with well-controlled primary sclerosing cholangitis can undergo ileal pouch-anal anastomosis surgery safely. PMID- 12576891 TI - Multiple hemorrhoidal ligation: a prospective, randomized trial evaluating a new technique. AB - PURPOSE: A modified anoscope was developed, with lateral apertures at the left lateral, right anterior, and right posterior quadrants, to enable synchronous exposure and ligation of all three internal hemorrhoids. Results were compared with those for conventional multiple ligation. METHODS: Postligation pain, complications, and outcomes were compared between synchronous ligation with the new anoscope (synchronous group) and three-quadrant ligation with a conventional anoscope with similar overall dimensions (conventional group). RESULTS: Twenty five patients were prospectively randomized to each group. Postligation pain and analgesic requirements were recorded up to 28 days, and postligation complications and outcomes were evaluated for a minimum of 6 months. Narcotic requirements were lower in the synchronous group, but this difference did not achieve statistical significance (P > 0.05, Student's t-test). Secondary hemorrhage occurred in 1 patient (4 percent) in the conventional group but resolved spontaneously. The synchronous group experienced significantly less pain during the ligation procedure and for 2 days afterward (P < 0.01, Wilcoxon's test). External hemorrhoidal thrombosis developed in 4 percent of the synchronous group and 12 percent of conventionally treated patients, all of whom responded to conservative treatment. Repeat ligation was required less often in the synchronous group (16 percent) than with conventional ligation (28 percent). Surgery (completion hemorrhoidectomy for external thrombosis) was necessary in one patient (4 percent) in each group. Anal stenosis developed in one patient in the synchronous group. CONCLUSION: The new anoscope provides improved exposure of all three internal hemorrhoids and permits optimal placement of the rubber bands; this may account for the decreased postligation pain and lower repeat ligation rates. Synchronous hemorrhoidal ligation is a less painful method of multiple hemorrhoidal ligation and may improve outcomes compared with conventional multiple ligation. PMID- 12576892 TI - Comparison of a novel liquid (Adcon-P) and a sodium hyaluronate and carboxymethylcellulose membrane (Seprafilm) in postsurgical adhesion formation in a murine model. AB - PURPOSE: Intra-abdominal administration of antiadhesive barriers may reduce the extent and severity of postoperative adhesions. This study aimed to compare the effectiveness of a novel liquid antiadhesive barrier with a conventional sheet (Seprafilm) antiadhesive barrier in a murine cecal abrasion model. METHODS: One hundred fifty Swiss-Wister mice underwent laparotomy and cecal abrasion and were randomly assigned to receive Adcon-P (n = 30) or Seprafilm (n = 30) or to a control group (n = 90). At postoperative Day 21, the mice underwent relaparotomy and complete adhesiolysis. An investigator who was blinded to the group assignment scored the extent of adhesion formation and the difficulty of adhesiolysis using a 6-point scale that ranged from 0 (no adhesions) to 5 (full thickness intestinal injury with adhesiolysis). Results are reported as median (range). RESULTS: Median adhesion scores in mice receiving Adcon-P (0 (range, 0 1)) and Seprafilm (1 (range, 0-3)) were lower than in mice in the control group (2 (range, 0-5); P < 0.0001 for both comparisons). In addition, the median adhesion score for the Adcon-P group was significantly lower than that of the Seprafilm group (P = 0.02). CONCLUSION: This study suggests that both Adcon-P and Seprafilm trade mark decrease the incidence of postoperative adhesions and the difficulty of adhesiolysis in the murine cecal abrasion model. However, Adcon-P appeared to be superior to Seprafilm. This agent is an attractive device that requires additional studies. PMID- 12576893 TI - p53, deleted in colorectal cancer gene, and thymidylate synthase as predictors of histopathologic response and survival in low, locally advanced rectal cancer treated with preoperative adjuvant therapy. AB - PURPOSE: Adjuvant therapy, either preoperatively or postoperatively, and modifications of surgery have been used to try to improve outcome of surgery for rectal cancer in regard to both local recurrence and survival. Assessment of prognosis in patients after resection is currently primarily based on clinicopathologic factors. These predict the subsequent behavior of the tumor only imperfectly. The aim of this study was to evaluate three potential molecular genetic markers of prognosis (p53, deleted in colorectal cancer gene, and thymidylate synthase) in Dukes Stage B and C low rectal tumors treated with adjuvant therapy and to determine whether they correlate with survival, local recurrence, or the pathologic response to adjuvant therapy (assessed by extent of tumor regression and tumor down-staging). METHODS: Sixty locally advanced low rectal tumors resected after preoperative chemoradiotherapy or radiotherapy alone were studied by immunohistochemical staining for p53, deleted in colorectal cancer gene, and thymidylate synthase. In addition, p53 gene mutations were sought by polymerase chain reaction-single-strand conformation polymorphism analysis. These results were correlated with survival, local recurrence, and pathologic response to adjuvant therapy. RESULTS: Lack of thymidylate synthase staining by immunohistochemistry was associated with tumor down-staging after preoperative chemoradiotherapy but not after radiotherapy or for these two combined groups. There was no correlation between p53, deleted in colorectal cancer gene, or thymidylate synthase immunohistochemical staining or between p53 polymerase chain reaction-single-strand conformation polymorphism and local recurrence or survival in locally advanced low rectal cancers treated with preoperative adjuvant therapies. CONCLUSION: Prediction of prognosis in patients with locally advanced low rectal cancers treated with preoperative adjuvant therapies continues to be problematic. Thymidylate synthase immunohistochemistry appears to be the most promising factor of those assessed in predicting tumor down-staging after preoperative chemoradiotherapy for locally advanced low rectal cancers. PMID- 12576894 TI - Risk of lymph node and distant metastases in patients with early invasive colorectal cancer classified as Haggitt's level 4 invasion: image analysis of submucosal layer invasion. AB - PURPOSE: Tumor invasion in patients with early invasive colorectal cancer has been classified into four levels proposed by Haggitt. Level 4 invasion into the submucosa has been defined as a risk factor for lymph node metastasis; however, the false-positive rate remains high. This study was designed to determine risk factors for lymph node and distant metastases in addition to Haggitt's Level 4 invasion. METHODS: Seventy-one of 142 patients with submucosa-invasive colorectal cancer underwent intestinal resection as an initial surgical treatment between 1975 and 2000. The subjects of this study were 65 of these 71 patients, all of whom were diagnosed as having Haggitt's Level 4 invasion. The depth, width, and area of submucosal invasion were measured with an image analyzer. RESULTS: Lymph node metastasis was noted in 11 (16.9 percent) of the 65 patients. There were no significant differences in the depth or area of submucosal invasion between node positive and node-negative patients. However, the width of submucosal invasion was significantly greater in node-positive than in node-negative patients (P = 0.001). When 5-mm-wide submucosal invasion was used as an indicator for intestinal resection, 37 patients were found to have indications for bowel resection, and 11 (29.7 percent) of the 37 had lymph node metastases. Distant metastasis was noted in five patients (7.7 percent). The depth, width, and area of submucosal invasion in patients with distant metastasis did not differ significantly from those without distant metastasis. CONCLUSION: Although further prospective investigation is required, the positive predictive value increases from 17 to 30 percent when the width of submucosal invasion is added to Haggitt's Level 4 as an indicator for bowel resection. PMID- 12576895 TI - Mesenteric phlebosclerosis: a new disease entity causing ischemic colitis. AB - PURPOSE: Nonthrombotic stenosis or occlusion of the mesenteric veins is a rare cause of intestinal ischemia. The aim of this study was to describe a new disease entity causing chronic ischemic colitis. METHODS: Seven patients were diagnosed as having mesenteric phlebosclerosis. All seven patients had calcifications in the small mesenteric veins and their intramural branches. No evidence of vasculitis or portal hypertension was recognized. None of the patients had a history of gastrointestinal disease or of prolonged drug use. We report clinical, laboratory, radiographic, endoscopic, and histopathologic findings. RESULTS: Clinical findings included abdominal pain and diarrhea of a gradual onset and chronic course. A positive fecal occult blood test and mild anemia were often found. The patients had linear calcifications and stenosis in the right colon, which were discovered by plain abdominal radiography and barium enema, respectively. Endoscopic findings included edematous, dark colored mucosa and ulcerations. Four patients underwent a subtotal colectomy because of persistent abdominal pain or ileus. The histopathologic findings were macroscopically characterized by a dark purple or dark brown colored colonic surface, the swelling and disappearance of plicae semilunares coli, and marked thickening of the colonic wall, while they were microscopically characterized by marked fibrous thickening of the venous walls with calcifications, marked submucosal fibrosis, deposition of collagen in the mucosa, and foamy macrophages within the vessel walls. CONCLUSIONS: These peculiar lesions have not previously been fully described. The cause and pathogenesis still remain unknown. We conclude that such lesions represent a new clinicopathologic disease entity and propose the term "idiopathic mesenteric phlebosclerosis." PMID- 12576896 TI - Prognostic significance of occult metastases in colon cancer. AB - PURPOSE: The purpose of this study was to determine the prognostic significance of occult lymph node metastases in colon cancer detected by cytokeratin 20 reverse transcription polymerase chain reaction. METHODS: Two hundred patients undergoing elective colonic resections were enrolled in the study. Lymph nodes from resected specimens were dissected fresh and assessed by both reverse transcription polymerase chain reaction and histopathology. Follow-up was undertaken for up to five years, and the major end point of death was recorded. Univariate survival analysis was performed by the log-rank method and the change in-estimate method was used to construct multivariate analysis models for the effect of cytokeratin 20 reverse transcription polymerase chain reaction lymph node status on overall survival. RESULTS: A total of 2,317 lymph nodes from 200 patients were assessed by both histopathology and cytokeratin 20 reverse transcription polymerase chain reaction. Forty-eight of 141 (34 percent) histologically lymph node-negative patients had evidence of occult metastases by cytokeratin 20 reverse transcription polymerase chain reaction. An interim analysis was performed at a median of 42 (range, 23-75) months. Cytokeratin 20 reverse transcription polymerase chain reaction lymph node status was a highly significant predictor of overall survival (P < 0.0001) on univariate analysis. In addition, the number of reverse transcription polymerase chain reaction-positive lymph nodes was a significant predictor of survival in the histologically lymph node-negative group (P < 0.0001) on univariate analysis. On multivariate analysis cytokeratin 20 reverse transcription polymerase chain reaction lymph node status had independent prognostic significance for overall survival (P = 0.021; hazard ratio = 2.7) and the number of cytokeratin 20 reverse transcription polymerase chain reaction-positive lymph nodes was a significant predictor of overall survival in the histologically lymph node-negative group (P = 0.005; hazard ratio = 1.1-11.1). CONCLUSION: Cytokeratin 20 reverse transcription polymerase chain reaction has potential as a clinically useful marker for staging colorectal cancer. Further follow-up is required, but if the current trends continue, a study of the effect of adjuvant therapy in patients with occult metastases detected by cytokeratin 20 reverse transcription polymerase chain reaction is indicated. PMID- 12576898 TI - Rectal hyposensitivity: prevalence and clinical impact in patients with intractable constipation and fecal incontinence. AB - PURPOSE: Blunted rectal sensation, or rectal hyposensitivity, has been reported anecdotally in patients with functional disorders of evacuation and continence. The purpose of this study was to determine the prevalence of rectal hyposensitivity and whether the finding of such an abnormality was associated with any clinical impact. METHODS: One thousand three hundred fifty-one patients, referred for anorectal physiologic investigation, were divided according to presenting symptoms into the following categories: constipation (subdivided into infrequency of and/or obstructed defecation), fecal incontinence (subdivided into passive, postdefecation, and urge incontinence), fecal incontinence and constipation, or "other." Rectal hyposensitivity was judged to be present when at least one of the sensory threshold volumes was elevated beyond the normal range (mean plus 2 standard deviations). The prevalence of rectal hyposensitivity was then calculated in each group and in relation to other investigations. RESULTS: Rectal hyposensitivity was present in 16 percent of patients, with males and females equally affected. Twenty-three percent of patients with constipation, 10 percent of patients with fecal incontinence, 27 percent of patients with incontinence associated with constipation, and only 5 percent of patients with other symptoms were found to have rectal hyposensitivity. In patients with obstructed defecation, rectal hyposensitivity was present in 33 percent with rectocele, 40 percent with intussusception, and 53 percent with no mechanical obstruction evident on evacuation proctography. CONCLUSION: Rectal hyposensitivity is common in patients with constipation and/or fecal incontinence and may thus be important in the etiology of such conditions. Although the clinical relevance of this physiologic abnormality is unknown, its presence may have implications regarding the management of hindgut dysfunction and particularly the selection of patients for surgery. PMID- 12576897 TI - Comparison of botulinum toxin injection and lateral internal sphincterotomy for the treatment of chronic anal fissure. AB - PURPOSE: Botulinum toxin injection into the internal anal sphincter has been shown to be an effective treatment for chronic anal fissure. A randomized, prospective trial was conducted to compare botulinum toxin with lateral internal anal sphincterotomy as definitive management for chronic anal fissure. METHODS: Patients diagnosed as having chronic anal fissure were randomly assigned to one of the two treatment arms. In the botulinum toxin group (n = 61), 20 to 30 U (approximately 0.3 U/kg) of type A botulinum toxin (Botox) was injected into the internal anal sphincter. The injection was repeated two months later if complete healing was not accomplished. Patients in the sphincterotomy group (n = 50) underwent lateral internal anal sphincterotomy. The same investigators evaluated the patients on postoperative/postinjection days 7 and 28, and then in a blinded manner at 2, 6, and 12 months. RESULTS: In the botulinum group, single injection resulted in complete healing in 45 of the 61 patients (73.8 percent) at the second month. Of the 16 failures, 6 patients refused further treatment, and 10 were treated with a second injection, which resulted in an overall healing rate of 86.9 percent (53/61) at 6 months. In the sphincterotomy group, the success rate was 82 percent (41/50) at day 28 and 98 percent (49/50) at the second month (P = 0.023 and P < 0.0001, respectively, compared with the botulinum group-single injection). At 6 months, 2 patients in the LIS group developed recurrences, and the healing rate was similar to that of the botulinum group (86.9 96.4 percent; P = 0.212). At 12 months, the success rate of the Botox group fell to 75.4 percent (46/61) with 7 recurrences, whereas it remained stable in the sphincterotomy group (94 percent, P = 0.008). Sphincterotomy was associated with a significantly higher complication rate (8 cases of anal incontinence none in the botulinum toxin group; P < 0.001). Full return to daily activities took significantly less time in the botulinum group (1 14.8 +/- 5.7 days; P < 0.0001). CONCLUSION: Although the healing rate of chronic anal fissure is considerably high with botulinum toxin injection with earlier recovery and less complications compared with sphincterotomy, it occasionally requires a repeat injection, and the healing is slower. The early (two months) and late (one year) healing rates are significantly higher in the sphincterotomy group, the two groups reaching similar healing rates only at six months. PMID- 12576899 TI - Management of late complications of pelvic radiation in the rectum and anus: a review. AB - PURPOSE: Radiotherapy has become one of the most important treatment modalities for human malignancy. Tumors affecting the organs of the pelvis are increasingly being irradiated for local treatment benefit, with the subsequent complication of anorectal injury of varying extent. The aim of this review is to determine how to manage the consequences of long-term effects of radiotherapy on the rectum and anus. METHODS: A comprehensive search of the literature with manual cross referencing was performed using the MEDLINE, PubMed, and Cochrane Databases. RESULTS: Long-term manifestations of injury caused by pelvic radiotherapy include abscess and fistula formation, stricture, mucus discharge, urgency, tenesmus, diarrhea, increased risk of cancer, and most commonly, bleeding. Most patients present with several symptoms; however, usually one symptom dominates. CONCLUSIONS: Many of these symptoms are self-limiting, and mucosal complications may often be treated by nonsurgical methods such as topical formalin application, endoscopic argon plasma coagulation, and hyperbaric oxygen therapy. Conservative measures have not been shown to be of benefit if symptoms persist. Structural abnormalities and septic complications are likely to require surgery. Modern techniques in the delivery of radiotherapy help minimize the likelihood of rectal complications. PMID- 12576900 TI - Combined use of preoperative provocative angiography and highly selective methylene blue injection to localize an occult small-bowel bleeding site in a patient with Crohn's disease: report of a case. AB - PURPOSE: Gastrointestinal bleeding in patients with Crohn's disease presents both a diagnostic and therapeutic challenge. The bleeding site may be difficult to localize preoperatively and multiple segments of gross disease can lead to uncertainty as to the precise source at the time of laparotomy. METHODS: We describe a patient with Crohn's disease and recurrent gastrointestinal bleeding in whom the combined use of provocative angiography and highly selective methylene blue injection was used preoperatively to accurately identify the site of hemorrhage and direct bowel resection. RESULTS: Provocative angiography identified the bleeding point in the jejunum. Methylene blue, which had been injected distally into the bleeding vessel during angiography, stained the bowel wall at the bleeding site. Segmental bowel resection was subsequently performed and no further bleeding occurred during the 18-month follow-up period. CONCLUSIONS: The combined use of provocative angiography and highly selective methylene blue injection may aid in the preoperative and intraoperative localization of occult bleeding sites in patients with Crohn's disease. This allows the bleeding lesion to be removed with a limited resection, thus preserving bowel length. PMID- 12576901 TI - Choriocarcinoma of the colon: report of a case and review of the literature. AB - Primary choriocarcinoma of the colon is a very rare tumor, with only six reported cases in the world literature, all but one of which was associated with an adjacent adenocarcinoma. This has led to the suggestion that colonic choriocarcinomas may arise from the more typical adenocarcinoma a process of further dedifferentiation. This article reviews the above cases and adds a further case from a 73-year-old male in whom no associated adenocarcinoma could be found despite careful postmortem examination. This finding gives support to the hypothesis that, rather than arising as a result of further dedifferentiation of an existing tumor, primary choriocarcinomas may also develop in the large intestine. PMID- 12576902 TI - Clostridium difficile infection--an unusual cause of refractory pouchitis: report of a case. AB - PURPOSE: Ileal pouch-anal anastomosis is the surgical procedure of choice for selected patients with severe ulcerative colitis. Pouchitis is a common complication of this procedure, with most cases responding to treatment with metronidazole, possibly with the addition of 5-aminosalicylic acid drugs and steroids. can frequently colonize the colon after treatment with broad-spectrum antibiotics, giving rise to diarrhea or colitis. The aim of this report was to describe the first case of -associated diarrhea manifest as pouchitis. METHODS: The management of refractory pouchitis in a 35-year-old female with toxin in the stool is described followed by a literature review of small-intestinal infection. RESULTS: Assays for toxin on stool sent during an episode considered to be caused by idiopathic chronic pouchitis were positive, and treatment with oral vancomycin was initiated. The patient responded with a reduction in bowel frequency to twice daily, a successful discontinuation of her antidiarrheal medication, and a rapid increase in weight. A subsequent stool assay was negative for the toxin. CONCLUSIONS: infection can complicate pouchitis in patients with an ileal pouch anal anastomosis and should be considered in patients who fail to respond to standard treatment, including metronidazole. In cases of refractory pouchitis, superadded infection with should be excluded before initiation of potent anti inflammatory drugs. PMID- 12576903 TI - Minimally invasive, endoscopically assisted colostomy can be performed without general anesthesia or laparotomy. AB - PURPOSE: Fecal diversion is frequently required in critically ill patients who may not be able to tolerate a laparotomy. Laparoscopic-assisted and trephine colostomies are alternative methods for colostomy without laparotomy, but require general anesthetic. The objective of this study was to evaluate the possibility of performing fecal diversion with the assistance of a colonoscope and without the additional morbidity of abdominal exploration or general anesthesia. METHODS: Patients were diverted using a colonoscope to identify a site of the sigmoid colon that could easily be approximated to the anterior abdominal wall as confirmed by transillumination of the abdominal wall. A small skin disc was then removed at this location and a loop colostomy was made. The colonoscope was also used as a guide to identify the proximal and distal limbs of the loop colostomy. Four patients were considered to be critically ill and local or regional anesthetic with sedation was used in these patients. RESULTS: A total of 15 patients were reviewed during the past five years. All 15 patients were successfully diverted using minimally invasive techniques with the aid of the colonoscope. Four of these patients were diverted using local or regional anesthetic without complication, thus avoiding the morbidity associated with a general anesthetic in critically ill patients. CONCLUSION: No complications related to this technique were noted in this five-year review. Endoscopically assisted colostomy is an acceptable method for fecal diversion without the need for laparotomy and can be accomplished using a local or regional anesthetic with sedation. PMID- 12576904 TI - Vacuum-assisted closure of a complex pilonidal sinus. AB - INTRODUCTION: The annual incidence of pilonidal sinus is approximately 26 per 100,000, of which complex pilonidal sinuses are the minority. Many different approaches have been described for managing simple cases. Treatment options for complex pilonidal sinus include excision with healing by secondary intention or plastic surgical procedures to obliterate the defect. Recently, vacuum-assisted closure has been used by plastic surgeons to facilitate healing of chronic or complicated wounds with particular success in treating pressure wounds overlying the sacrum. METHODS: A patient with a complex pilonidal sinus was managed with excision and vacuum-assisted closure. RESULTS: A large tissue defect after radical excision healed relatively quickly when the subatmospheric pressure dressing was applied. The patient was discharged with the vacuum pump and change of dressings for alternate days. CONCLUSION: Vacuum-assisted closure of a complex pilonidal sinus shortened the length of hospital stay and the need for further surgery and provided a cosmetically acceptable result. PMID- 12576905 TI - Randomizing the dead. PMID- 12576906 TI - Exercise training for heart failure patients improves respiratory muscle endurance, exercise tolerance, breathlessness, and quality of life. AB - PURPOSE: Increased respiratory muscle endurance and peak oxygen consumption (VO(2peak)) induced by respiratory muscle training support the relationship between respiratory muscle function and exercise capacity in patients with heart failure. This raises the question whether exercise-training results in increased respiratory muscle function contributing to an increased exercise tolerance, a decreased perception of breathlessness, and an improved quality of life. METHODS: Prospective cohort analysis was completed on 24 patients with New York Heart Association (NYHA) Class III heart failure [18 men, 6 women; aged = 64 (SD 7.9) years; percent ejection fraction (%EF) = 24.0 (SD 7.8)]. Maximal sustainable ventilatory capacity (MSVC), submaximal and peak exercise responses, perception of breathlessness, and quality of life were measured before (baseline) and after (end of study) 12 weeks of exercise training. RESULTS: As a result of exercise training, VO(2peak) (P=.01) and MSVC (P<.001) increased, with MSVC contributing to a larger proportion of the variability for VO(2peak) at study completion (r=0.57 vs 0.42). Although stroke volume did not increase beyond exercise at 25 W and did not change with exercise training, ventilation decreased during exercise (P<.05), perception of breathing difficulty (P<.05) was reduced, and quality of life was enhanced (P=.008). CONCLUSIONS: Despite no increase in cardiac output and stroke volume, respiratory muscle endurance improved with exercise training, contributing to increased exercise capacity, decreased breathlessness, and decreased perception of breathlessness. Practical implications can include less frequent rest periods and fatigue, greater confidence, maintenance of independence, and enhanced quality of life. PMID- 12576907 TI - Four-week multidisciplinary cardiac rehabilitation produces similar improvements in exercise capacity and quality of life to a 10-week program. AB - BACKGROUND: Cardiac rehabilitation (CR) is widely accepted as beneficial for patients with myocardial infarction (MI) and coronary artery bypass graft (CABG). A need exists to evaluate how different formats of delivery can best meet CR service demands. METHODS AND RESULTS: Cardiac patients (n = 60) were randomly assigned to either a standard 10-week (30 sessions) or a 4-week (20 sessions) multifactorial rehabilitation program. Patients underwent exercise testing using the Bruce protocol before, immediately after, and then 6 months after CR. Patients also completed the SF-36 quality of life questionnaire and the Hospital Anxiety and Depression scale at each time point. Compared with pre-CR, exercise time and metabolic equivalents attained were significantly increased, and heart rate significantly decreased both immediately (P<.05) and 6 months after CR (P<.05) in both groups, with no between-group differences. Significant improvements (P<.05) in energy, pain, and general health were reported after CR, and in energy and emotional and social well-being at 6 months after CR. No differences were seen between the groups. CONCLUSIONS: Cardiac rehabilitation after MI and CABG significantly improved exercise capacity and general health and well-being. No significant differences were detected between groups undergoing a 10-week or 4-week course. These preliminary data suggest that shortened courses of CR may be beneficial to cardiac patients and such courses may also facilitate more widespread use of CR. PMID- 12576908 TI - Short Form-36 Health Survey as an evaluation tool for cardiac rehabilitation programs: is it appropriate? PMID- 12576909 TI - Assessing health-related quality of life: is it important when evaluation the effectiveness of cardiac rehabilitation? PMID- 12576910 TI - Is the theory of planned behavior a useful framework for understanding exercise adherence during phase II cardiac rehabilitation? AB - PURPOSE: This study evaluated the theory of planned behavior (TPB) as a framework for understanding exercise adherence during phase II cardiac rehabilitation (CR). METHODS: A total of 215 patients completed a baseline questionnaire that included the TPB constructs and past exercise. Exercise adherence was measured via program attendance during phase II CR. RESULTS: Hierarchic regression analyses indicated that attitude, subjective norm, and perceived behavioral control (PBC) explained 30% of the variance in exercise intention, with attitude, subjective norm, and PBC each making significant unique contributions to intention. Furthermore, exercise intention explained 12% of the variance in exercise adherence. Finally, the behavioral, normative, and control beliefs provided novel information concerning why patients in phase II CR hold certain attitudes, subjective norms, PBC, and exercise intentions. CONCLUSION: Results of the present study provide evidence that the TPB is a useful framework for understanding exercise intentions and adherence during phase II CR. PMID- 12576911 TI - Predictors of women's exercise maintenance after cardiac rehabilitation. AB - PURPOSE: Less than 50% of persons who participate in cardiac rehabilitation (CR) programs maintain an exercise regimen for as long as 6 months after completion. This study was conducted to identify factors that predict women's exercise following completion of a CR program. METHODS: In this prospective, descriptive study, a convenience sample of 60 women were recruited at completion of a phase II CR program. Exercise was measured using a heart rate wristwatch monitor over 3 months. Predictor variables collected at the time of the subjects' enrollment were age, body mass index, cardiac functional status, comorbidity, muscle or joint pain, motivation, mood state, social support, self-efficacy, perceived benefits or barriers, and prior exercise. RESULTS: Of women, 25% did not exercise at all following completion of a CR program and only 48% of the subjects were exercising at 3 months. Different predictors were found of the various dimensions of exercise maintenance. Predictors of exercise frequency were comorbidity and instrumental social support. Instrumental social support was the only predictor of exercise persistence. Comorbidity was the only predictor of exercise intensity. The only predictor of the total amount of exercise was benefits or barriers. CONCLUSIONS: Interventions aimed at increasing women's exercise should focus on increasing their problem-solving abilities to reduce barriers to exercise and increase social support by family and friends. Because comorbidity was a significant predictor of exercise, women should be encouraged to use exercise techniques that reduce impact on muscles and joints (eg, swimming) or exercising for short periods several times a day. PMID- 12576912 TI - Women and exercise maintenance: challenges and opportunities. PMID- 12576913 TI - Body image and depressive symptoms as correlates of self-reported versus clinician-reported physiologic function. AB - PURPOSE: This study examined the relationships between physiologic function, depressive symptoms, and body image among maintenance cardiac rehabilitation participants. Physiologic function was operationalized as both functional status and functional capacity. METHODS: Participants were 72 men (mean age = 67.3 years) all of whom had experienced a traumatic cardiac event (ie, myocardial infarction, valve replacement surgery, coronary artery bypass graft surgery, percutaneous transluminal coronary angioplasty), and had completed some type of physician-supervised acute cardiac rehabilitation (ie, phase I and phase II). Measures of body image (social physique anxiety and body appearance satisfaction), self-reported functional status, clinician-reported functional capacity (ie, V0(2) and peak power), and depressive symptoms were collected. RESULTS: Hierarchic multiple regression analyses revealed that both functional capacity and functional status explained significant variance in social physique anxiety (R(2) = 0.11, P<.05 and R(2) = 0.18, P<.05, respectively), whereas only functional status was a significant predictor of body appearance satisfaction (R(2) = 0.37, P<.01). Contrary to our hypotheses, depressive symptoms were not significantly related to either psychosocial or physiologic indices of functional well-being. CONCLUSIONS: Both patient perceptions of functional status and clinical measures of functional capacity are important aspects of psychosocial well-being among cardiac patients. PMID- 12576915 TI - Prognostic factors in localized aggressive non-Hodgkin's lymphoma. AB - To identify the prognostic factors that specifically predict survival rates of patients with localized aggressive non-Hodgkin's lymphoma (NHL), a retrospective study including 118 patients with clinical stage I and II NHL treated at the Institute of oncology, Istanbul University between 1989 and 1998 was conducted. Patients were treated either with radiotherapy alone, radiotherapy and adjuvant chemotherapy, or chemotherapy (with or without adjuvant radiotherapy). The 5-year disease-free survival (DFS) and overall survival rates were calculated, and univariate and multivariate analyses were performed to identify the significance of various prognostic factors such as gender, age, performance status, stage (I versus II), B symptoms, extranodal involvement, gastrointestinal tract disease, erythrocyte sedimentation rate, bulky disease, histologic grade, serum lactate dehydrogenase level, serum beta2-microglobulin level, serum albumin level, treatment regimen, remission status, and the International Prognostic Index risk groups, which may have an influence on the outcome of patients with NHL. The overall 5-year survival rate was 52% with a median follow-up of 30 months. The complete response rate was 68%, and the 5-year DFS of complete responders was 70%. Cox multivariate regression analysis showed that incomplete response, low serum albumin, bulky disease (>10 cm), and high grade histology were the pretreatment factors associated with shorter survival. When remission status was included in the model, the attainment of a complete response was the major determinant of long-term survival; however, low albumin level was still a significant adverse predictor for survival in multivariate analysis. These factors need to be evaluated for analyzing the outcome of treatment and to identify better therapeutic strategies. PMID- 12576914 TI - A randomized, controlled trial comparing long-term and short-term exercise in patients with chronic obstructive pulmonary disease. AB - PURPOSE: To compare the effects of short-term (3 months) and long-term (18 months) involvement in an exercise program on self-reported disability and physical function in patients with chronic obstructive pulmonary disease (COPD). METHODS: A total of 140 patients with COPD were studied in a randomized, single blinded clinical trial. Self-reported disability and physical function were assessed using a 21-item questionnaire, a 6-minute walk, timed stair climb, and an overhead task. RESULTS: At the completion of the trial, participants in the long-term intervention reported 12% less disability than those in the short-term intervention (adjusted mean with 95% confidence interval, 1.53 (1.43-1.63) versus 1.71 (1.61 to 1.81) units, respectively; P=.016), walked 6% farther during 6 minutes (1,815.0 [1,750.4-1,879.6] vs 1,711.5 [1,640.7-1,782.3] feet, respectively), climbed steps 11% faster (11.6 [11.0-12.2] vs 12.9 [12.3-13.5] seconds, respectively), and completed an overhead task 8% faster (46.8 [44.4 49.2] vs 50.4 [47.8-53.0] seconds, respectively) than those in the short-term intervention. CONCLUSION: An 18 month exercise program results in greater improvements in self-reported disability and physical function in patients with COPD when compared with a 3-month exercise program. As such, long-term exercise should be recommended for all patients with COPD. PMID- 12576916 TI - Lymphoma of the nasal cavity and paranasal sinuses: treatment and outcome of early-stage disease. AB - The records of 23 patients diagnosed and treated at the Massachusetts General Hospital for extranodal non-Hodgkin's lymphoma of the paranasal sinus and nasal cavity were reviewed. The majority of patients were Ann Arbor stage I and approximately evenly divided in T1 or T2 (n = 10) and T3 or T4 (n = 13). Eight patients had nasal-type NK/T cell and 15 patients had diffuse large B-cell lymphoma (DLBCL). The patients with nasal-type NK/T cell lymphoma predominately involved the nasal cavity (5/8), whereas the DLBCL more often had the paranasal sinuses as the primary site (12/15). All patients received radiation as part of their treatment. Only three patients received chemotherapy as part of their initial treatment for three cycles using a cyclophosphamide, doxorubicin, vincristine, and prednisone-based regimen. By coincidence, the estimated overall survival (OS) and disease-free survival rates for both 5 and 10 years were all the same for all analyses. The OS for the entire group at 10 years was 78%. Significant prognostic factors were Ann Arbor stage IEA versus IIEA ( p = 0.0001) and T stage with (T1 or T2) versus (T3 or T4) (p = 0.0243). Combining Ann Arbor stage and T stage created a highly significant prognostic variable (IEA & [T1 or T2], IEA & [T3 or T4], IIEA & [T1 or T2], IIEA & [T3 or T4]) at p = 0.0001, regardless of site or histology. Patients with local-regional disease appear to be well controlled with radiation alone, but distant failure remains a problem. A combined-modality approach with local-regional radiation and systemic chemotherapy is recommended for these patients. PMID- 12576917 TI - Reduced maintenance of complete protection from emesis for women during chemotherapy cycles. AB - Women have a significantly higher rate of chemotherapy-induced emesis. We observed that gender was the important predict factor for reducing the maintenance of complete protection from emesis during chemotherapy cycles. Four hundred patients were enrolled in one of two arms in a crossover fashion. Arm A: three 8-mg doses of ondansetron were given intravenously at 4-hour intervals plus dexamethasone 20 mg intravenously from the start of chemotherapy, followed by dexamethasone 5 mg intravenously every 12 hours. Arm B: as in arm A but with three 8-mg doses of ondansetron intravenously given at 24-hour intervals substituted for ondansetron intravenously given at 4-hour intervals. Rates for complete protection from vomiting/nausea through days 1 to 6 were 69.7%/58.4% for Arm A, and 71.2%/60.9% for Arm B, and 69.0%/60.0% for the first chemotherapy cycle. Risk factors for vomiting and nausea included gender and cisplatin dosage. Complete control of vomiting/nausea for patients without emesis during previous cycle(s) was maintained at 91.4%/86.6%, 91.1%/85.2%, 93.9%/89.9%, 94.7%/92.6%, and 94.9%/94.9% during the second through sixth cycles of chemotherapy, respectively. Complete protection from vomiting and nausea was significantly reduced for female patients (p = 0.048 and p = 0.0004, during the second cycle, and p = 0.0006 and p = 0.0008, during the third through sixth cycles, respectively). The results suggest that women had less maintenance for complete protection from both vomiting and nausea during chemotherapy cycles. PMID- 12576918 TI - Impact of intraoperative radiation on postoperative and disease-specific outcome after pancreatoduodenectomy for adenocarcinoma: a propensity score analysis. AB - For periampullary cancer,intraoperative radiation therapy (IORT) administered to the site with the highest locoregional recurrence risk carries the rationale to improve tumor control. An IORT effect on survival remains unclear. IORT impact on postoperative outcomes after pancreatectomy for adenocarcinoma was analyzed, with a specific attempt to correct for the nonrandom IORT treatment assignment, and to account for treatment group imbalances in the interpretation of outcome differences. A propensity-score-adjusted analysis, based on variable selection by logistic regression, was used to rebalance treatments. Between 1989 and 1999, 61 patients underwent partial or total pancreatectomy for a primary periampullary adenocarcinoma at the City of Hope National Medical Center. Diagnoses included pancreatic (n = 36), duodenal (n = 11), ampullary (n = 10), and bile duct cancer (n = 4). Thirty patients received IORT to the resection area, with a median dose of 15 Gy (range: 10-20), followed by postoperative external beam radiation (n = 24). Mortality was 0%, the complication rate 61%. Of 33 patients with a documented recurrence, 6 had an isolated locoregional recurrence only (1 IORT versus 5 no IORT, = 0.05); the systemic recurrence pattern differed as well (IORT 94%, no IORT 67%; = 0.04). IORT had no significant impact on hospital stay (overall median: 17 days), disease-free survival (16 months), and overall survival (23 months) when adjusted for those most relevant variables reflecting IORT treatment group assignment propensity. After adjustment for relevant propensity factors, IORT was not linked to a significantly increased risk for complications, hospital stay, or survival hazard. The recurrence pattern may be affected in some patients, but systemic recurrences predominate. We continue to explore IORT in combination with systemic chemotherapy. PMID- 12576920 TI - Paraneoplastic Raynaud phenomenon and idiopathic thrombocytopenic purpura in non small-cell lung cancer. AB - The association of Raynaud phenomenon with malignancy has rarely been reported. Idiopathic thrombocytopenic purpura is also an infrequent complication of solid tumors. We report a 62-year-old man who sought treatment for severe Raynaud phenomenon requiring medical therapy and was subsequently diagnosed to have non small-cell lung cancer. After chemotherapy and radiotherapy, his cancer was in complete remission and the Raynaud phenomenon resolved. Severe thrombocytopenia later developed for which no apparent cause was found, and which was responsive to steroid therapy. This heralded relapse of his lung cancer. This is a case of lung cancer with two rare paraneoplastic syndromes. PMID- 12576919 TI - Induction chemotherapy with gemcitabine and oxaliplatin for locally advanced cervical carcinoma. AB - Induction chemotherapy followed by surgery, particularly with newer agents or combinations, remains to be explored in locally advanced cervical cancer. Gemcitabine cisplatin is a very active combination for this tumor, therefore we explored the activity of gemcitabine in combination with oxaliplatin. Ten untreated patients with histologic diagnosis of cervical carcinoma and staged as IB2 to IIIB were treated with 3 21-day courses of oxaliplatin 130 mg/m day 1 and gemcitabine 1,250 mg/m days 1 and 8 followed by locoregional treatment with either surgery or concomitant chemoradiation. Response and toxicity were evaluated at the end of chemotherapy. All patients were evaluable. The overall clinical response rate was 80%, being complete in 3 patients (30%) and partial in 5 (50%). Seven (70%) patients underwent surgery, and three (30%) had chemoradiation as definitive treatment. Hematologic toxicity was moderate, with leukopenia grades III-IV in 17 and 0%; granulocytopenia grades III-IV in 23 and 3%, respectively. Eight patients had grade I oropharyngeal toxicity. At a median follow-up of 11 months (range: 10-12), all patients are disease free. Gemcitabine oxaliplatin is a very active and well-tolerated combination for locally advanced cervical cancer. PMID- 12576921 TI - Surgical resection after preoperative chemoradiotherapy benefits selected patients with unresectable pancreatic cancer. AB - Simultaneous chemoradiation is used in unresectable pancreatic cancer for palliation. It is not known if the use of adjuvant surgery will benefit this group of patients. From November 1991 to September 1998, 47 patients with unresectable pancreatic cancer were treated with simultaneous preoperative radiation therapy (45 Gy) and chemotherapy. Chemotherapy followed three different protocols: cisplatin, 5-fluorouracil +/- paclitaxel; cisplatin, 5-fluorouracil (protracted infusion); and docetaxel and gemcitabine. Whipple pancreatoduodenectomy was performed 1 month after the end of radiation in patients selected for resection. Twenty-three unresectable tumors after preoperative treatment (47%) received an additional dose (10-12 Gy) of radiotherapy using intraoperative or external radiation therapy. Twelve patients (26%) were considered to have clinically resectable tumors after the preoperative treatment. Nine patients had surgery (19% of the total number of patients), and 2 of them had complete pathologic response. After chemoradiation, two patients died of pneumonia and gastrointestinal bleeding, respectively, and another two patients died in the postoperative period. Local recurrence was observed in 22% of the patients and 57% had distant metastases. Three-year survival rates for patients with unresectable and resectable tumors was 0% (median survival 10 months) and 48% (median survival 23 months), respectively (p = 0.0004). Preoperative treatment with chemotherapy and radiotherapy in patients with unresectable pancreatic cancer is feasible. In some patients, the tumor can be resected, and in addition some cases of complete pathologic response were found. Long-term survivors were observed in the group of resected tumors. More effective chemotherapy regimens are needed because the majority of the patients died of metastatic disease. PMID- 12576922 TI - Phase II study of paclitaxel and carboplatin in patients with advanced gastric cancer. AB - 5-Fluorouracil-based combination chemotherapy is commonly used in patients with advanced gastric cancer, but results with such therapy are fairly modest. Evaluation of newer agents is therefore required in this disease. Paclitaxel has shown promising activity as a single agent in gastric cancer. In vitro, paclitaxel exhibits sequence-dependent synergy with platinum compounds against gastric cancer. This study was conducted to evaluate the efficacy and toxicity of combination carboplatin and paclitaxel in patients with advanced gastric cancer. Twenty-seven patients with measurable or evaluable advanced gastric cancer were enrolled on the study from April 1996 to July 2000. Patients were treated with paclitaxel 200 mg/m intravenously during 3 hours followed by carboplatin at projected area under the curve 5 mg x h x ml (as per the Calvert formula). Twenty six patients were assessable for toxicity, and 25 patients were assessable for objective response. Nine of the 27 enrolled patients had a major response for an objective response rate of 33% (95% CI 0.17-0.54) by intention-to-treat analysis. The median response duration was 4.9 months (95% CI 2.8-7.3), and median survival was 7.5 months. The 1-year survival rate was 23%. One hundred seventeen courses were administered with a median of four courses per patient administered. The major toxicity was neutropenia, with grade III to IV neutropenia observed in 9 patients (33%) and neutropenic fever in only 1 patient. Grade III peripheral neuropathy developed in two patients, and grade III myalgia and grade III fatigue developed in one patient each. There were no treatment-related deaths. The combination of carboplatin and paclitaxel is a highly tolerable, regimen with activity comparable to that of other regimens in advanced gastric cancer. This regimen needs to be further evaluated in combination with other agents and as a component of multimodality therapy in gastric cancer. PMID- 12576923 TI - Parathyroid hormone-related protein and hypercalcemia in patients with metastatic melanoma: case report and review. AB - Hypercalcemia associated with malignancy has been attributed to osteolytic processes secondary to bony metastases and to humoral factors causing increased bone resorption and decreased renal excretion of calcium. Parathyroid hormone related protein (PTH-rP) is a humoral factor that has been associated with hypercalcemia in renal cell carcinoma, squamous cell carcinoma, and bladder carcinoma. Hypercalcemia does occur in patients with melanoma; however, few studies have reported on hypercalcemia in these patients, and even fewer have described a direct connection to PTH-rP. We here report a patient with stage IV malignant melanoma presenting with severe hypercalcemia associated with elevated PTH-rP levels. Immunohistochemistry showed strong expression of PTH-rP in biopsy of the patient's subcutaneous masses. In addition, we found a 4.9% incidence of hypercalcemia in 1,146 consecutive patients treated for metastatic melanoma at the Surgery Branch of the National Cancer Institute between January 1, 1988 and March 31, 2000. Thus, PTH-rP may play a significant role in severe hypercalcemia in patients with metastatic melanoma. The discovery of PTH-rP and relevant literature will also be reviewed. PMID- 12576924 TI - Radiation therapy combined with cis-diammine-glycolatoplatinum (Nedaplatin) and 5 fluorouracil for untreated and recurrent esophageal cancer. AB - From January 1999 to November 2000, a total of 24 esophageal cancer patients (17 untreated and 7 recurrent cases) were treated with radiation therapy (60-70 Gy) combined with cis-diammine-glycolatoplatinum (Nedaplatin) (80-120 mg/body) and 5 fluorouracil (5-FU) (500-1,000 mg/body/24 h, continuous infusion for 5 days). Grade III leukocytopenia was observed in 6 (25%) of the patients. Grade III and IV thrombocytopenia was observed in one patient each. The 1-year and 2-year survival rates for definitively irradiated patients were 59% and 39%, respectively, and for patients with postoperative recurrence 69% and 69%, respectively. High-dose radiation combined with Nedaplatin and 5-FU is a safe and effective method for treating esophageal cancer. PMID- 12576925 TI - Decreased response to paclitaxel versus docetaxel in HER-2/neu transfected human breast cancer cells. AB - Taxanes are effective in the treatment of metastatic breast cancer. Docetaxel has been shown to be more potent than paclitaxel in inducing bcl-2 phosphorylation and apoptosis and is clinically active in some paclitaxel-resistant breast tumors. HER-2/neu overexpression has been shown to correlate with resistance to hormonal therapy as well as chemotherapy. Using a HER-2/neu transfected MCF-7 human breast cancer cell line, we investigated the role of HER-2/neu overexpression on resistance to paclitaxel and docetaxel treatment. A control vector transfected MCF-7 human breast cancer cell line (MCF/neo) and a HER-2/neu transfected MCF-7 line (MCF/18) were treated with various concentrations of docetaxel or paclitaxel. Cell number was assessed using the MTT tetrazolium dye assay. In the control vector transfected MCF/neo cell line, paclitaxel and docetaxel gave similar dose-dependent growth inhibition ( p = 0.175). In HER 2/neu transfected MCF/18 cells, docetaxel treatment resulted in a dose-dependent inhibition similar to that seen in MCF/neo cells. Paclitaxel, however, gave significantly less growth inhibition than docetaxel in the HER-2/neu overexpressing MCF/18 cells (p = 0.0003). These data suggest that HER-2/neu overexpression may contribute to paclitaxel resistance. In contrast, the cytotoxic effects of docetaxel in these breast carcinoma cells are not affected by HER-2/neu expression. Therefore, docetaxel may be the preferred taxane therapy in HER-2/neu overexpressing breast tumors. PMID- 12576926 TI - Protracted radiotherapy treatment duration in medulloblastoma. AB - From 1970 to 1997, 63 patients with medulloblastoma were treated with craniospinal irradiation followed by a posterior fossa boost. There were 38 males and 25 females with a median age of 9 years (range, 8 months to 53 years). Stage was T1-T3a in 50 (79%) and M0 in 38 patients (60%) according to the Chang staging system. Gross total resection of the primary tumor was achieved in 33 (52%) and median posterior fossa dose was 54 Gy, with 55 (87%) receiving > or =50 Gy. Median radiotherapy treatment duration was 49 days (range, 30-104 days) with 35 patients (56%) completing radiotherapy in <50 days. The most common reasons for a protracted radiotherapy treatment duration > or =50 days were hematologic toxicity (46%) and use of <1.6 Gy fraction size per day (29%). Chemotherapy was used in 22 (35%). Median follow-up time was 10.8 years (range, 2-28.5 years). The 5- and 10-year freedom from progression rates were 58% +/- 13% and 50% +/- 13%, respectively, whereas the 5- and 10-year posterior fossa control rates were 61% +/- 12% and 54% +/- 13%, respectively. On multivariate analysis, age > or =3 years, M0 status, > or =50 Gy PFB dose, radiotherapy treatment duration <50 days, and use of chemotherapy correlated with better freedom from progression and posterior fossa control rates. The 5- and 10-year freedom from progression rates were 67% +/- 15% and 64% +/- 16%, respectively, for patients with radiotherapy treatment duration <50 days and were 42% +/- 20% and 29% +/- 18%, respectively, for duration > or =50 days ( p= 0.0026, log-rank test). The 5- and 10-year posterior fossa control rates were 70% +/- 15% and 70% +/- 15%, respectively, for radiotherapy treatment duration <50 days and 46% +/- 20% and 33% +/- 19%, respectively, for duration > or =50 days ( p= 0.0037, log-rank test). In addition to age > or =3 years, M0 stage, use of adjuvant chemotherapy, and posterior fossa dose > or =50 Gy, our findings also reveal that radiotherapy treatment duration <50 days has a favorable prognostic outcome in patients with medulloblastoma. PMID- 12576927 TI - Radiation enhancement by gemcitabine-mediated cell cycle modulations. AB - The purpose of this study was to investigate the exact dose dependency and time dependency of the radiation-enhancing effect of gemcitabine (2',2'difluoro desoxycytidine [dFdC]) in in vitro experiments (HeLa cells: cancer of the uterine cervix, #4197 cells: oropharyngeal squamous cell carcinoma), and to correlate this effect with the underlying changes in cell cycle distribution. Cell viability was determined fluorometrically after exposure to dFdC (0-20.0 micro mol/l), irradiation (0-37.5 Gy), and both modalities. Combining both therapies, cells were exposed to dFdC (0-10.0 micro mol/l) for 24 hours before further treatment and irradiated (0-30 Gy) immediately afterwards with or without removal of dFdC. For cell cycle analysis by flow cytometry, cells were irradiated (0-40 Gy) or treated with dFdC (0.012-1.0 micro mol/l, 24-48 hours). Additionally, cells were exposed to dFdC (2.0 micro mol/l, 0-4 hours). Cell cycle kinetics were evaluated using bromodeoxyuridine (BrdU) (10 micro mol/l) S-phase labeling, given either 30 minutes before or in the last hour of dFdC treatment (2.0 micro mol/l, 0-6 hours). The fluorometric assay revealed that dFdC enhances radiation-induced cytotoxicity at marginally toxic or nontoxic concentrations (<37 nmol/l). Radiation resulted in the anticipated G2/M arrest already at 2 Gy. DFdC induced concentration and exposure time-dependent cell cycle changes that were better resolved using BrdU, demonstrating a pronounced S-phase arrest already at 12 nmol/l. BrdU-pulse labeling revealed that the cell cycle block occurred at the G1/S boundary. Our data reconfirm the already known radiation enhancement, the S phase specific activities of dFdC, and the relevance of the synchronized progression of cells through the S-phase with regard to the radiosensitizing properties of low-dose dFdC. However, we could demonstrate that before progressing in the S-phase, cells were blocked and partially synchronized at the more radiosensitive G1/S boundary. Furthermore, cells progressing past the block might accumulate proapoptotic signals caused by both radiation and dFdC, which will also results in cell death. PMID- 12576928 TI - Reduction in radiation dose to lung and other normal tissues using helical tomotherapy to treat lung cancer, in comparison to conventional field arrangements. AB - The purpose of this study was to determine whether the use of tomotherapy in the treatment of non-small-cell lung cancer (NSCLC) has the potential to reduce radiation dose to normal tissues, in particular, the lungs, esophagus, and spinal cord, as compared with standard radiotherapy. Five patients with anatomically or physiologically inoperable stage III NSCLC were studied, representing a variety of tumor sizes and locations. For each patient, two treatment plans were generated. One was developed using conventional field arrangements (CFA), and the other for tomotherapy. Using dose-volume histogram reduction techniques, including mean normalized dose (NTDmean), V20, and effective uniform dose (EUD), the normal tissue doses for CFA and tomotherapy plans for a given fixed tumor dose were compared. In addition, the maximum tumor doses possible for a given level of mean normalized lung dose were computed and compared for the CFA and tomotherapy plans. The gross tumor volumes in the five patients studied ranged from 13.5 to 87.1 cm. The tumor dose distributions, determined by EUD and minimum dose, were similar for both CFA and tomotherapy plans, as intended. In all cases, the NTDmean of both lungs was significantly reduced using tomotherapy planning (range: 10-53% reduction, mean: 31%). The volume of lung receiving more than 20 Gy was also reduced in all cases using tomotherapy (range: 17-37% reduction, mean: 22%). For a constant lung NTDmean, it is shown that it should be possible to increase tumor dose to up to 160 Gy in certain patients with tomotherapy. The dose to the spinal cord and esophagus was also reduced in all cases with tomotherapy planning, compared with plans generated using conventional field arrangements. Both tomotherapy, and to a lesser extent conventional three dimensional conformal radiotherapy, have the potential to significantly decrease radiation dose to lung and other normal structures in the treatment of NSCLC. This has important implications for dose escalation strategies in the future. PMID- 12576929 TI - Multiple primary malignant neoplasms: case report and a comprehensive review of the literature. AB - A case is presented of an elderly patient with synchronous ureteral/bladder/urethral transitional cell carcinoma and prostatic adenocarcinoma. In a subsequent review of 1,104,269 cancer patients in the literature, the reported prevalence of multiple primary malignant neoplasms (MPMN) varies between 0.734% and 11.7%. It appears that MPMN might occur more frequently than can be explained on the basis of random chance. As expected, the incidence of developing MPMN is noted to rise with increasing age. In addition, the preponderance of men with MPMN is caused primarily by the high frequency of prostatic cancer. Current studies and research need to address the potential of older cancer patients being at higher risk of second primaries. PMID- 12576931 TI - Metastatic breast carcinoma presenting as a large pulmonary embolus: case report and review of the literature. AB - Involvement of the pulmonary vasculature by carcinoma of the breast typically occurs in the form of microscopic tumor emboli involving the small arteries, arterioles, or capillaries. Obstruction of a large pulmonary artery by a tumor embolus has not been reported. We describe a patient with a history of breast carcinoma diagnosed 5 years previously who sought treatment for dyspnea and a large mass in the right pulmonary artery suggestive of a pulmonary embolus. After failure of both systemic and intraarterial thrombolytic therapy, a biopsy of the mass was obtained, which revealed adenocarcinoma of the breast. Systemic chemotherapy with doxorubicin and cyclophosphamide was initiated and resulted in the complete resolution of her symptoms. PMID- 12576930 TI - Phase II study of Taxol combined With ifosfamide and carboplatin in the treatment of stage IIIb-IV non-small-cell lung cancer. AB - The objective of the present study was to evaluate the activity and the toxicity of an original combination of paclitaxel (Taxol), ifosfamide, and carboplatin in patients with stage IIIB-IV non-small-cell lung cancer (NSCLC). Sixty-one patients with previously untreated stage IIIB-IV NSCLC were enrolled by five institutions. Paclitaxel was given at the dose of 200 mg/m iv in 3 hours, ifosfamide (with mesna) at the dose of 3 g/m and carboplatin at an area under the curve 5, on day 1, every 21 days for a total of six cycles in responding or stabilized patients. Among the 59 patients evaluable for response, 2 complete remissions and 25 partial remissions were achieved for an overall response rate of 45.7% (95% CI = 32.7-59.2). According to an intention-to-treat analysis, the response rate was 44.2%. Thirteen patients had a stable disease, whereas 19 progressed. The median time to progression was 7.7 months (range: 1-18), whereas the median overall survival was 10 months (range: 1-30+). The 1-year survival rate was 43%. Hematologic toxicity was exceptionally mild, and peripheral neurologic toxicity of grade III was experienced by only three patients. There was one toxic death. This original triplet regimen based on paclitaxel, ifosfamide, and carboplatin has proved active, safe, and easy to deliver on an outpatient basis for patients with advanced NSCLC. Randomized studies both versus carboplatin-paclitaxel and other triplets are clearly warranted. PMID- 12576932 TI - Regression of lung lesions in Hodgkin's disease by antibiotics: case report and hypothesis on the etiology of Hodgkin's disease. AB - In this article, we propose that the pathogenesis of Hodgkin's disease is similar to the one of crown gall tumors in plants. Here a natural exchange of genetic material from (oncogenic plasmids) to plant cells induces malignant tumors in dicotyledons. The "crown gall" hypothesis for Hodgkin's disease would explain the clinical observations of a bacterial infection the behavior as a malignant tumor. The clinical consequence of this hypothesis is that antibiotic treatments of very early Hodgkin's disease may be successful before the genetic exchange between prokaryotic and eukaryotic cells has taken place. This "crown gall" hypothesis is testable (1) by looking for bacterial DNA sequences in Reed-Sternberg and Hodgkin's cells, and (2) by antibiotic treatments of Hodgkin's patients. In this communication we show a regression of Hodgkin's disease in the lung by prolonged treatment with ciprofloxacin and clarithromycin. PMID- 12576933 TI - Safety and activity of docetaxel and trastuzumab in HER2 overexpressing metastatic breast cancer: a pilot phase II study. AB - We conducted a pilot phase II trial of trastuzumab administered concurrently with docetaxel in women with HER2-overexpressing advanced breast cancer. Twenty-five women with HER2-positive (3+ by immunohistochemistry = 16, 2+ = 9) metastatic breast cancer received docetaxel (75 mg/m every 3 weeks for 6 cycles) and trastuzumab (4 mg/kg loading dose, 2 mg/kg weekly thereafter). Twenty-three patients (92%) had visceral metastatic involvement. Twenty-three patients had received prior chemotherapy as part of adjuvant (18), metastatic (2), and both (3) treatment. The number of cycles administered was 121 (median 6, range 1-6). Symptomatic cardiotoxicity (GIII) occurred in one patient. The most common grade GIII/IV toxicity was neutropenia (80% of the cycles), although febrile neutropenia did not occur. No other GIII/IV toxicities were observed. Response rate was 70% (1 complete response and 15 partial responses) in 23 evaluable patients. The combination of docetaxel and trastuzumab is well tolerated and has clinically meaningful antitumor activity. PMID- 12576934 TI - Oral doxifluridine plus leucovorin in metastatic colorectal cancer: randomized phase II trial with intravenous 5-fluorouracil plus leucovorin. AB - This phase II study was designed to evaluate the efficacy and toxicities of oral doxifluridine plus leucovorin as a randomized trial with those of intravenous 5 fluorouracil (5-FU) plus leucovorin in previously untreated metastatic colorectal cancer (CRC). Patients with metastatic CRC were randomized in either group A (oral doxifluridine 1,000 mg/m /d plus leucovorin 30 mg/d on days 1 to 7 and 15 to 21 of each cycle), or group B (intravenous 5-FU 400 mg/m /d plus leucovorin 20 mg/m /d on days 1-5 of each cycle), with the cycles repeated every 4 weeks. Between July 1998 and May 2000, 77 patients were enrolled (38 in group A and 39 in group B). Response rates were 23.7% (95% CI, 11-42%) in group A, and 15.4% (95% CI, 0-25%) in group B on an intent-to-treat analysis. The median response durations of the two groups were similar with 5.6 months in group A and 5.5 months in group B. Progression-free survival and overall survival were 5.4 months and 14.9 months in group A; 4.7 months and 19.5 months in group B. Toxicities in both groups were generally mild and reversible. This study shows that a combination of oral doxifluridine plus leucovorin can be active and safe as a first-line treatment for patients with metastatic CRC. PMID- 12576935 TI - Severe toxicity related to the 5-fluorouracil/leucovorin combination (the Mayo Clinic regimen): a prospective study in colorectal cancer patients. AB - The Mayo Clinic regimen of leucovorin 20 mg/m followed immediately by 5 fluorouracil 425 mg/m administered for 5 consecutive days every 4 weeks is commonly used in the treatment of colorectal cancer. This study was aimed at prospectively determining the incidence and pattern of severe toxicity associated with this regimen. We evaluated prospectively 243 patients with colorectal cancer treated in our department with the Mayo Clinic regimen for the incidence of severe toxicity (defined as toxicity requiring hospitalization). Of the 243 patients, 32 (13%) were hospitalized for chemotherapy-related toxicity. Major toxicities included neutropenic fever in 21 (9%), grade III/IV mucositis in 25 (10%) and grade III/IV diarrhea in 20 (8%). There were five (2%) treatment related deaths. Female patients exhibited a higher incidence of severe toxicity (18%) and toxic death (4/105) than did male patients (9% and 1/138, respectively). Elderly patients (> or =70 years) had a higher incidence of severe toxicity than younger patients did (24% versus 7%, < 0.001). Toxic death occurred in 4 of 89 patients aged 70 years or more compared to 1 of 154 in younger patients. Most episodes of severe toxicity (56%) and toxic deaths (4/5) were observed after the first cycle. We conclude that the Mayo Clinic regimen can be associated with severe toxicity, usually occurring after the first cycle. Female gender and advanced age predict severe toxicity; therefore, dose reduction in high-risk patients should be considered, especially during the first cycle. PMID- 12576936 TI - Impaired mitochondrial function induced by serum from septic shock patients is attenuated by inhibition of nitric oxide synthase and poly(ADP-ribose) synthase. AB - OBJECTIVE: The purpose of this study was to determine the role of nitric oxide and poly(ADP-ribose) synthase on impaired mitochondrial function in septic shock. DESIGN: Human umbilical vein endothelial cells were incubated with serum from ten healthy controls, 20 patients with septic shock, and seven critically ill patients who were not septic. The experiment was repeated after pretreatment with 3-aminobenzamide, a poly(ADP-ribose) synthase inhibitor, or N(G)-methyl-L arginine, a nonspecific nitric oxide synthase inhibitor. MEASUREMENTS: Mitochondrial respiration was measured using a modified MTT (3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay. SETTING: Research laboratory. MAIN RESULT: Endothelial cell mitochondrial respiration was significantly depressed by septic serum and averaged 61% +/- 6% of control values (p <.05). Incubation with septic serum as compared with control serum also significantly decreased cellular adenosine triphosphate levels (6.7 +/- 1.2 nM vs. 13.5 +/- 1.9 nM, p<.01). The level of mitochondrial respiration in endothelial cells exposed to septic serum did not correlate with arterial lactate concentration but was correlated with both cardiac output (r(s) =.52, p<.05) and mixed venous oxygen saturation (r(s) =.61, p<.05). Pretreatment with N(G)-methyl L-arginine significantly increased mitochondrial respiration in endothelial cells treated with septic serum from 63% +/- 6% of normal to 88% +/- 6% (p <.05) of normal values. Similarly, pretreatment with 3-aminobenzamide increased mitochondrial respiration in endothelial cells treated with septic serum from 64% +/- 6% to 100% +/- 4% (p <.01) of normal values. Endothelial cells incubated with serum from nonseptic critically ill patients did not demonstrate a significant decrease in mitochondrial respiration. CONCLUSION: In vitro mitochondrial respiration was significantly depressed by septic serum. The addition of N(G) methyl-L-arginine, a nitric oxide synthase inhibitor, and 3-aminobenzamide, a blocker of the poly(ADP-ribose) synthase pathway, significantly attenuated this suppression. These data suggest that nitric oxide and poly(ADP-ribose) synthase activation may play an important role in the inhibition of mitochondrial respiration in septic shock. PMID- 12576937 TI - Outcome benefit of intensive insulin therapy in the critically ill: Insulin dose versus glycemic control. AB - OBJECTIVES: Maintenance of normoglycemia with insulin reduces mortality and morbidity of critically ill patients. Here we report the factors determining insulin requirements and the impact of insulin dose vs. blood glucose control on the observed outcome benefits. DESIGN: A prospective, randomized, controlled trial. SETTING: A 56-bed predominantly surgical intensive care unit in a tertiary teaching hospital. PATIENTS AND INTERVENTION: A total of 1,548 patients were randomly assigned to either strict normalization of blood glucose (80-110 mg/dL) with insulin infusion or the conventional approach, in which insulin is only given to maintain blood glucose levels at 180-200 mg/dL. MEASUREMENTS AND MAIN RESULTS: It was feasible and safe to achieve and maintain blood glucose levels at <110 mg/dL by using a titration algorithm. Stepwise linear regression analysis identified body mass index, history of diabetes, reason for intensive care unit admission, at-admission hyperglycemia, caloric intake, and time in intensive care unit as independent determinants of insulin requirements, together explaining 36% of its variation. With nutritional intake increasing from a mean of 550 to 1600 calories/day during the first 7 days of intensive care, normoglycemia was reached within 24 hrs, with a mean daily insulin dose of 77 IU and maintained with 94 IU on day 7. Insulin requirements were highest and most variable during the first 6 hrs of intensive care (mean, 7 IU/hr; 10% of patients required >20 IU/hr). Between day 7 and 12, insulin requirements decreased by 40% on stable caloric intake. Brief, clinically harmless hypoglycemia occurred in 5.2% of intensive insulin-treated patients on median day 6 (2-14) vs. 0.8% of conventionally treated patients on day 11 (2-10). The outcome benefits of intensive insulin therapy were equally present regardless of whether patients received enteral feeding. Multivariate logistic regression analysis indicated that the lowered blood glucose level rather than the insulin dose was related to reduced mortality (p <.0001), critical illness polyneuropathy (p <.0001), bacteremia (p =.02), and inflammation (p =.0006) but not to prevention of acute renal failure, for which the insulin dose was an independent determinant (p =.03). As compared with normoglycemia, an intermediate blood glucose level (110-150 mg/dL) was associated with worse outcome. CONCLUSION: Normoglycemia was safely reached within 24 hrs and maintained during intensive care by using insulin titration guidelines. Metabolic control, as reflected by normoglycemia, rather than the infused insulin dose, was related to the beneficial effects of intensive insulin therapy. PMID- 12576938 TI - Multicenter, double-blind, placebo-controlled study of the use of filgrastim in patients hospitalized with pneumonia and severe sepsis. AB - OBJECTIVE: To determine the safety and efficacy of filgrastim (r-metHuG-CSF) in combination with intravenous antibiotics to reduce the rate of mortality in patients with pneumonia and sepsis. DESIGN: This study was multicenter, double blind, and randomized. SETTING: Intensive care units PATIENTS Adult patients with bacterial pneumonia, either acquired or nosocomial, as confirmed by chest radiograph and positive culture or Gram-negative stain, and severe sepsis, defined as sepsis-induced hypotension or organ dysfunction. INTERVENTIONS: Standard antibiotic therapy with or without filgrastim (300 microg/day) or placebo administered as a 30-min intravenous infusion. The study drug was started within 24 hrs of enrollment and was continued for 5 days or until the white blood cell count reached >75.0 x 10(9) cells/L. MEASUREMENTS AND MAIN RESULTS: The primary end point was the occurrence of mortality through day 29; secondary end points included occurrence of subsequent organ dysfunction, time to discharge from intensive care unit, number of days on mechanical ventilatory support, and time to death. Study-related observations were recorded through day 10 and included vital signs, onset of organ dysfunction, clinical laboratory variables, and adverse events. Filgrastim increased the white blood cell count to a median peak of 31.7 x 10(9) cells/L from a baseline of 12.3 x 10(9) cells/L. The two groups were well matched and did not differ significantly with regard to severe adverse events, time to death, occurrence of end-organ dysfunction, days of intensive care unit hospitalization, or days on mechanical ventilatory support. Mortality was low in both treatment groups; the mortality rate in patients with adult respiratory distress syndrome was similar between the two groups. CONCLUSIONS: The addition of filgrastim to the antibiotic and supportive care treatment of patients with pneumonia complicated by severe sepsis appeared to be safe, but not efficacious in reducing mortality rates or complications from this infection. PMID- 12576939 TI - Nursing activities score. AB - OBJECTIVES: The instruments used for measuring nursing workload in the intensive care unit (e.g., Therapeutic Intervention Scoring System-28) are based on therapeutic interventions related to severity of illness. Many nursing activities are not necessarily related to severity of illness, and cost-effectiveness studies require the accurate evaluation of nursing activities. The aim of the study was to determine the nursing activities that best describe workload in the intensive care unit and to attribute weights to these activities so that the score describes average time consumption instead of severity of illness. DESIGN: To define by consensus a list of nursing activities, to determine the average time consumption of these activities by use of a 1-wk observational cross sectional study, and to compare these results with those of the Therapeutic Intervention Scoring System-28. SETTING: A total of 99 intensive care units in 15 countries. PATIENTS: Consecutive admissions to the intensive care units. INTERVENTION: Daily recording of nursing activities at a patient level and random multimoment recording of these activities. RESULTS: A total of five new items and 14 subitems describing nursing activities in the intensive care unit (e.g., monitoring, care of relatives, administrative tasks) were added to the list of therapeutic interventions in Therapeutic Intervention Scoring System-28. Data from 2,041 patients (6,451 nursing days and 127,951 multimoment recordings) were analyzed. The new activities accounted for 60% of the average nursing time; the new scoring system (Nursing Activities Score) explained 81% of the nursing time (vs. 43% in Therapeutic Intervention Scoring System-28). The weights in the Therapeutic Intervention Scoring System-28 are not derived from the use of nursing time. CONCLUSIONS: Our study suggests that the Nursing Activities Score measures the consumption of nursing time in the intensive care unit. These results should be validated in independent databases. PMID- 12576940 TI - Acute oxygenation response to inhaled nitric oxide when combined with high frequency oscillatory ventilation in adults with acute respiratory distress syndrome. AB - OBJECTIVE: To prospectively evaluate the oxygenation effect of inhaled nitric oxide (INO) delivered during high-frequency oscillatory ventilation in adult patients with the acute respiratory distress syndrome and oxygenation failure. DESIGN Prospective, clinical study. SETTING: Intensive care unit of a university teaching hospital. PATIENTS: A total of 23 adults (14 women, 9 men, 44.9 +/- 17.5 yrs, Acute Physiology and Chronic Health Evaluation II score of 28.6 +/- 7.1) with acute respiratory distress syndrome (lung injury score, 3.5 +/- 0.4) with Fio2 of > or = 0.6 and mean airway pressure of >or=28 cm H2O. INTERVENTIONS: INO was initiated at a dose of 5 ppm, and subsequently titrated according to a protocol, to determine the dose (5, 10, or 20 ppm) resulting in the greatest increase in Pao2/Fio2. Blood gas measurements were obtained 10-15 mins after initiation or any increase in INO dosage to assess the effect on Pao2/Fio2. MEASUREMENTS AND MAIN RESULTS: Arterial blood gases and ventilator settings were recorded at four time points: during conventional ventilation just before initiating high-frequency oscillatory ventilation, during high-frequency oscillatory ventilation just before initiating INO, after 30 mins on the optimal dose of INO, and 8-12 hrs after starting INO. Oxygenation index ([Fio2 x mean airway pressure x 100]/Pao2) and Pao2/Fio2 ratios were calculated at the same time intervals. At 30 mins after INO initiation, 83% of patients had a significant increase in blood oxygen tension, defined as > or = 20% increase in Pao2/Fio2. The mean change in Pao2/Fio2 at 30 mins was 38%. In these 19 patients, Pao2/Fio2 was highest at 20 ppm in four patients, at 10 ppm in eight patients, and at 5 ppm in seven patients. Compared with baseline measurements, Pao2/Fio2 improved significantly at both 30 mins (112 +/- 59 vs. 75 +/- 32, p=.01) and 8-12 hrs after INO initiation (146 +/- 52 vs. 75 +/- 32, p<.0001). In addition, oxygenation index was reduced at 8-12 hrs compared with baseline measurements (26 +/- 13 vs. 40 +/- 17, p=.08). CONCLUSIONS: INO delivered at doses of 5 to 20 ppm during high-frequency oscillatory ventilation increases Pao2/Fio2 and may be a safe and effective rescue therapy for patients with severe oxygenation failure. PMID- 12576941 TI - Bacteremic sepsis in intensive care: temporal trends in incidence, organ dysfunction, and prognosis. AB - OBJECTIVE: To determine whether changes have occurred at our center in the incidence, patterns of organ dysfunctions, prognostic factors, and case-fatality rate of bacteremic sepsis. DESIGN: Retrospective comparative study (1984-1988 vs. 1994-1997). SETTING: University hospital surgical intensive care unit. PATIENTS: A total of 372 critically ill patients with sepsis and positive blood cultures. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: In the 1984-1988 study, 176 patients developed bacteremic sepsis (3.2 per 100 admissions), with a fatality rate of 35% at 28 days, whereas in the 1994-1997 study, 196 patients had bacteremic sepsis (4.3 per 100 admissions), with a 28-day fatality rate of 37%. The frequency of primary bacteremia increased from 21% to 47% (p <.001), paralleled by an increase in the proportional frequency of Gram-positive microorganisms. In 1984-1988, 69% (n = 120) of patients had at least one organ dysfunction, compared with 80% (n = 156) in 1994-1997 (p =.02). The prevalence of pulmonary and cardiac dysfunction increased. The case- fatality rate of septic shock remained high (69% vs. 68%). For both cohorts, the two strongest predictors of mortality remained the Acute Physiology and Chronic Health Evaluation II score at the onset of sepsis and the number of evolving organ dysfunctions. CONCLUSION: The fatality rate of bacteremic sepsis remained constant over the study period, despite an increased incidence of bacteremia and associated organ dysfunction. Continued efforts need to be directed toward the prevention of bacteremic sepsis, given the magnitude and poor prognosis of this syndrome. PMID- 12576942 TI - Iron and iron-related proteins in the lower respiratory tract of patients with acute respiratory distress syndrome. AB - OBJECTIVE: An increased oxidative stress in the lower respiratory tract of individuals with acute respiratory distress syndrome is considered to be one mechanism of lung injury in these patients. Cell and tissue damage resulting from an oxidative stress can ultimately be the consequence of a disruption of normal iron metabolism and an increased availability of catalytically active metal. Using bronchoalveolar lavage fluid, we quantified concentrations of iron and iron related proteins in the lower respiratory tract in patients with acute respiratory distress syndrome and healthy volunteers. DESIGN: A clinical study to quantify iron and iron-related proteins in the lower respiratory tract in patients with acute respiratory distress syndrome and healthy volunteers. PATIENTS: We studied 14 patients with acute respiratory distress syndrome and 28 healthy volunteers. MAIN RESULTS: Comparable to previous investigation, protein, albumin, and cytokine concentrations in the bronchoalveolar lavage fluid were significantly increased in acute respiratory distress syndrome patients. The concentrations of total and nonheme iron were also increased in the lavage fluid of patients. Concentrations of hemoglobin, haptoglobin, transferrin, transferrin receptor, lactoferrin, and ferritin in the bronchoalveolar lavage fluid were all significantly increased in acute respiratory distress syndrome patients. CONCLUSIONS: We conclude that bronchoalveolar lavage fluid indices reflect a disruption of normal iron metabolism in the lungs of acute respiratory distress syndrome patients. Increased concentrations of available iron in acute respiratory distress syndrome may participate in catalyzing oxidant generation destructive to the tissues of the lower respiratory tract. However, increased metal availability is also likely to elicit an increased expression of transferrin receptor, lactoferrin, and ferritin in the lower respiratory tract which will function to diminish this oxidative stress. PMID- 12576943 TI - Effectiveness of direct-current cardioversion for treatment of supraventricular tachyarrhythmias, in particular atrial fibrillation, in surgical intensive care patients. AB - OBJECTIVE: To evaluate primary success rate and effectiveness of direct-current cardioversion in postoperative critically ill patients with new-onset supraventricular tachyarrhythmias. DESIGN: Prospective intervention study. SETTING: Twelve-bed surgical intensive care unit in a university teaching hospital. PATIENTS: Thirty-seven consecutive, adult surgical intensive care unit patients with new-onset supraventricular tachyarrhythmias without previous history of tachyarrhythmias. INTERVENTIONS: Direct-current cardioversion using a monophasic, damped sinus-wave defibrillator. Energy levels used were 50, 100, 200, and 300 J for regular supraventricular tachyarrhythmias (n = 6) and 100, 200, and 360 J for irregular supraventricular tachyarrhythmias (n = 31). MEASUREMENTS AND MAIN RESULTS: None of the patients was hypoxic, hypokalemic, or hypomagnesemic at onset of supraventricular tachyarrhythmia. Direct-current cardioversion restored sinus rhythm in 13 of 37 patients (35% primary responders). Most patients responded to the first or second direct-current cardioversion shock. Only one of 25 patients requiring more than two direct current cardioversion shocks converted into sinus rhythm. Primary responders were significantly younger and demonstrated significant differences in arterial Po2 values at onset of supraventricular tachyarrhythmias compared with nonresponders. At 24 and 48 hrs, only six (16%) and five (13.5%) patients remained in sinus rhythm, respectively. CONCLUSIONS: In contrast to recent literature, direct current cardioversion proved to be an ineffective method for treatment of new onset supraventricular tachyarrhythmias and, in particular, atrial fibrillation with a rapid ventricular response in surgical intensive care unit patients. PMID- 12576944 TI - Time course of hemoglobin concentrations in nonbleeding intensive care unit patients. AB - OBJECTIVES: To evaluate the time course of hemoglobin concentrations in nonbleeding intensive care unit patients. DESIGN: Prospective, observational study. SETTING: Multidisciplinary (medicosurgical) department of intensive care. PATIENTS: Ninety-one patients with no evidence of recent or active blood loss, no history of hematologic disease or chronic renal failure, and no need for extracorporeal epuration techniques. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Data collection included primary diagnoses, Acute Physiology and Chronic Health Evaluation II and sepsis-related organ failure assessment scores, signs of sepsis, 24-hr fluid balance, and hemoglobin concentrations. For the entire intensive care unit stay, the fall in hemoglobin concentrations (calculated from the mean of individual slopes of hemoglobin concentrations over time) averaged 0.52 +/- 0.69 g/dL/day. For the 33 patients who stayed in the intensive care unit for >3 days, this decline was larger for the first 3 days than for subsequent days (0.66 +/- 0.84 g/dL/day vs. 0.12 +/- 0.29 g/dL/day; p<.01). After the third intensive care unit day, the change in hemoglobin concentrations was inversely related to the severity of the disease, as reflected by the Acute Physiology and Chronic Health Evaluation II and the sepsis-related organ failure assessment scores. Hemoglobin concentrations decreased by 0.44 +/- 0.70 g/dL/day in the nonseptic and 0.68 +/- 0.66 g/dL/day in the septic patients (p =.13). After the third intensive care unit day, hemoglobin concentrations continued to decrease in the septic patients but not in the nonseptic patients (-0.29 +/- 0.19 vs. 0.006 +/- 0.3 g/dL/day; p=.0016). The fall in hemoglobin concentrations was not significantly related to the fluid balance. The volume of blood drawn daily for laboratory studies was 40.3 +/- 15.4 mL: 49.0 +/- 11.3 mL in the septic patients and 36.7 +/- 14.9 mL in the nonseptic patients (p =.04). CONCLUSIONS: Hemoglobin concentrations typically decline by >0.5 g/dL/day during the first days of intensive care unit stay in nonbleeding patients. Beyond the third day, hemoglobin concentrations can remain relatively constant in nonseptic patients but continue to decrease in septic patients, as well as patients with high sepsis related organ failure assessment or Acute Physiology and Chronic Health Evaluation II scores. These observations may help in the interpretation of hemoglobin concentrations in critically ill patients. PMID- 12576945 TI - Effect of alveolar recruitment maneuver in early acute respiratory distress syndrome according to antiderecruitment strategy, etiological category of diffuse lung injury, and body position of the patient. AB - OBJECTIVE: To assess how the level of positive end-expiratory pressure (PEEP) (antiderecruitment strategy), etiological category of diffuse lung injury, and body position of the patient modify the effect of the alveolar recruitment maneuver (ARM) in acute respiratory distress syndrome (ARDS). DESIGN: Prospective clinical trial. SETTING: Medical intensive care unit at a tertiary hospital. PATIENTS: Forty-seven patients with early ARDS, including 19 patients from our preliminary study. INTERVENTION: From baseline ventilation at a tidal volume of 8 mL/kg and PEEP of 10 cm H2O, the ARM (a stepwise increase in the level of PEEP up to 30 cm H2O with a concomitant decrease in the magnitude of tidal volume down to 2 mL/kg) was given with (ARM + PEEP, n = 20) or without (ARM only, n = 19) subsequent increase of PEEP to 15 cm H2O. In eight other patients, PEEP was increased to 15 cm H2O without a preceding ARM (PEEP only). MEASUREMENTS AND RESULTS: In all three groups, Pao2 was increased by the respective intervention (all p<.05). In the ARM-only group, Pao2 at 15 mins after intervention was lower than Pao2 immediate after intervention (p =.046). In the ARM + PEEP group, no such decrease in Pao2 was observed, and Pao2 at 15, 30, 45, and 60 mins after intervention was higher than in the ARM-only group (all p<.05). Compared with the PEEP-only group, Pao2 of the ARM + PEEP group was higher immediately after intervention and at the later time points (all p <.05). Compared with patients with ARDS associated with direct lung injury (pulmonary ARDS), patients with ARDS associated with indirect lung injury (extrapulmonary ARDS) showed a greater increase in Pao2 (27 +/- 21% vs. 130 +/- 112%; p=.002) and a greater decrease in radiologic scores (1.0 +/- 2.4 vs. 3.4 +/- 1.5; p=.005) after the ARM. The increase in Pao2 induced by the ARM was greater for patients in the supine position than for patients in the prone position (61 +/- 82% vs. 21 +/- 14%; p=.028). Consequently, Pao immediately after the ARM was similar in the two groups of patients in different positions. CONCLUSIONS: After the ARM, a sufficient level of PEEP is required as an antiderecruitment strategy. Pulmonary ARDS and extrapulmonary ARDS may be different pathophysiologic entities. An effective ARM may obviate the need for the prone position in ARDS at least in terms of oxygenation. PMID- 12576946 TI - Multicenter evaluation of a human monoclonal antibody to Enterobacteriaceae common antigen in patients with Gram-negative sepsis. AB - OBJECTIVE: To evaluate in Gram-negative sepsis patients the human monoclonal immunoglobulin M antibody (MAB-T88) directed at the enterobacterial common antigen which is a specific surface antigen closely linked to lipopolysaccharide and shared by all members of the Enterobacteriaceae family of Gram-negative bacteria. DESIGN: Prospective, randomized, double-blinded, placebo-controlled, multicenter trial. SETTING: Thirty-three academic medical centers in the United States. PATIENTS: Patients were entered with a clinical diagnosis of sepsis, the presence of either shock or multiple organ dysfunction, and presumptive evidence for Gram-negative infection. INTERVENTIONS: Patients received a single intravenous infusion, over 30 mins, of either 300 mg of MAB-T88 formulated in albumin, or placebo (albumin). MEASUREMENTS AND MAIN RESULTS: The primary analysis group was prospectively identified as those patients with documented evidence of an infection with bacteria of the family Enterobacteriaceae at any site. The primary end point was survival within the first 28 days. A total of 826 patients were enrolled with 55% (n = 455) in the primary analysis group. There were no significant differences between the intervention and control primary analysis group study groups for sites of infection, severity of illness, underlying medical conditions, adequacy of antibiotic or surgical treatment, or other baseline variables except for a higher frequency of chronic renal failure in the MAB-T88 group (4.4% vs. 1.3%, p=.051). The average Acute Physiology and Chronic Health Evaluation II scores were 26.8 +/- 8.6 (mean +/- sd) in the MAB T88-treated group and 26.5 +/- 8.3 in the placebo-treated group (p =.72). There was no significant difference between MAB-T88- and placebo-treated groups during the first 28-day all-cause mortality in the primary analysis group (34.2% vs. 30.8%, p=.44) or in all 826 patients enrolled (37.0% vs. 34.0%, p=.36). On subset analysis, the use of MAB-T88 was not associated with significant mortality trends. More adverse events were seen with the use of MAB-T88 in the bacteremic enterobacterial common antigen group (p <.05). CONCLUSIONS: Use of the human monoclonal antibody, MAB-T88, did not improve the mortality in patients with presumed Gram-negative sepsis or in those patients with proven enterobacterial common antigen infections. No subset trends were identified that would support further investigation of this agent in sepsis. PMID- 12576948 TI - Physiologic data acquisition system and database for the study of disease dynamics in the intensive care unit. AB - OBJECTIVE: To describe a real-time, continuous physiologic data acquisition system for the study of disease dynamics in the intensive care unit. DESIGN: Descriptive report. SETTING: A 16-bed pediatric intensive care unit in a tertiary care children's hospital. PATIENTS: A total of 170 critically ill or injured pediatric patients. INTERVENTIONS: None. MAIN OUTCOME MEASURES: None. RESULTS: We describe a computerized data acquisition and analysis system for the study of critical illness and injury from the perspective of complex dynamic systems. Both parametric (1 Hz) and waveform (125-500 Hz) signals are recorded and analyzed. Waveform data include electrocardiogram, respiration, systemic arterial pressure (invasive and noninvasive), central venous pressure, pulmonary arterial pressure, left and right atrial pressures, intracranial pressure, body temperature, and oxygen saturation. Details of the system components are explained and examples are given from the resultant physiologic database of signal processing algorithms and signal analyses using linear and nonlinear metrics. CONCLUSIONS: We have successfully developed a real-time, continuous physiologic data acquisition system that can capture, store, and archive data from pediatric intensive care unit patients for subsequent time series analysis of dynamic changes in physiologic state. The physiologic signal database generated from this system is available for analysis of dynamic changes caused by critical illness and injury. PMID- 12576947 TI - Determinants of postintensive care unit mortality: a prospective multicenter study. AB - OBJECTIVE: Six to 25 percent of patients discharged alive from the intensive care unit (ICU) die before hospital discharge. Although this post-ICU mortality may indicate premature discharge from a full ICU or suboptimal management in the ICU or ward, another factor may be discharge from the ICU as part of a decision to limit treatment of hopelessly ill patients. We investigated determinants of post ICU mortality, with special attention to this factor. DESIGN: Prospective, multicenter, database study. SETTING: Seven ICUs in or near Paris, France. PATIENTS: A total of 1,385 patients who were discharged alive from an ICU after a stay of > or = 48 hrs; 150 (10.8%) died before hospital discharge. Decisions to withhold or withdraw life-sustaining treatments were implemented in the ICUs in 80 patients, including 47 (58.7%) who died before hospital discharge. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: In the univariate analysis, post-ICU mortality was associated with advanced age, poor chronic health status, severe comorbidities, severity and organ failure scores (Simplified Acute Physiology Score II, sepsis-related organ failure assessment, and Logistic Organ Dysfunction at admission and at ICU discharge), decisions to withhold or withdraw life-sustaining treatments, and Omega score (reflecting ICU resource utilization and length of ICU stay). Multivariate stepwise logistic regression identified five independent determinants of post-ICU mortality: McCabe class 1 (odds ratio, 0.388 [95% confidence interval, 0.26-0.58]), transfer from a ward (odds ratio, 1.89 [95% confidence interval, 1.27-2.80]), Simplified Acute Physiology Score II score at admission >36 (odds ratio, 1.57 [95% confidence interval, 1.6-2.33]), decisions to withhold or withdraw life-sustaining treatments (odds ratio, 9.64 [95% confidence interval, 5.75-16.6]), and worse sepsis-related organ failure assessment score at discharge (odds ratio, 1.11 [95% confidence interval, 1.03 1.18] per point). CONCLUSIONS: More than 10% of ICU survivors died before hospital discharge. Determinants of post-ICU mortality included variables reflecting patient status before and during the ICU stay. However, the most powerful predictor of post-ICU mortality was the decision to withhold or withdraw life-sustaining treatments in the ICU, suggesting that the decision has been made not to use the unique services of the ICU for these patients. PMID- 12576949 TI - Hyponatremia after hip arthroplasty may be related to a translocational rather than to a dilutional mechanism. AB - OBJECTIVE: Postoperative hyponatremia is a frequent metabolic disturbance that may cause life-threatening complications. It results from both a positive electrolyte-free water (EFW) balance and an antidiuretic hormone release. During surgery, intracellular solutes may leak out of cells because of an increased membrane permeability leading to increased osmolality, cellular water shift, and redistribution hyponatremia, a concept coined the sick cell syndrome. Because of release of osmotically active solutes, plasma or urinary osmolar gap should increase. Therefore, we tested the hypothesis that postoperative hyponatremia may be related to a translocational mechanism evidenced by a postoperative increase of the osmolar gap rather than to a positive EFW balance. SETTING: An anesthesiology department in a 1,200-bed university hospital. DESIGN: A 5-month prospective observational study. SUBJECTS: Thirty-three consecutive patients undergoing elective hip arthroplasty under general anesthesia. They were divided into two groups whether the postoperative plasma sodium concentration decrease was > or = 2 mmol/L (group 1) or <2 mmol/L (group 2). MEASUREMENTS: Plasma sodium concentration ([Na+]p) and plasma osmolality were measured before induction of anesthesia and at skin closure. Osmolality was calculated at the same times. Plasma osmolar gap (OG(p)) was calculated as the difference between measured and calculated osmolality. Postoperative urinary osmolar gap (OG(u)) was calculated in the same way. EFW balance was calculated as the ratio of (infused EFW - excreted urinary EFW) to total body water. RESULTS: In 33 patients, a significant [Na+]p decrease of -2.0 was observed. No relationship was demonstrated between EFW balance and perioperative [Na+]p variation (r =.28; p=.12). A relationship was observed between perioperative OG(p) variation and perioperative [Na+]p variation (r =.74; p<.0001). In the 19 group 1 patients, [Na+]p decreased by -3.0 mmol/L. EFW balance did not differ between group 1 and group 2 patients. No statistical relationship was observed between EFW balance and perioperative [Na+]p variation in group 1 (r =.20; p=.40) and in group 2 (r =.43; p=.14). OG(p) increased only in group 1 but not in group 2 patients, and postoperative OG(u) was greater in group 1 than in group 2 patients. A relationship was observed between perioperative OG(p) variation and perioperative [Na+]p variation in group 1 (r =.53; p=.02) but not in group 2 (r =.32; p=.26). CONCLUSION: Hyponatremia after hip arthroplasty may not be related to a positive EFW balance. The postoperative increase of the OG(p) and the greater postoperative OG(u) in patients developing postoperative hyponatremia suggest the release of osmotically active solutes leading to cellular water shift from intracellular to extracellular spaces. These data may support the clinical relevance of the sick cell syndrome in the postoperative context. PMID- 12576951 TI - Medical futility: predicting outcome of intensive care unit patients by nurses and doctors--a prospective comparative study. AB - OBJECTIVE: First, to assess the pattern of the prediction of intensive care unit patients' outcome with regard to survival and quality of life by nurses and doctors and, second, to compare these predictions with the quality of life reported by the surviving patients. DESIGN: Prospective opinion survey of critical care providers; comparison with follow-up for survival, functional status, and quality of life. SETTING: Six-bed medical intensive care unit subunit of a 1,000-bed tertiary care, university hospital. PATIENTS: All patients older than 18 yrs, admitted to the medical intensive care unit for >24 hrs over a 1-yr period (December 1997 to November 1998). INTERVENTIONS: Daily judgment of eventual futility of medical interventions by nurses and doctors with respect to survival and future quality of life. Telephone interviews with discharged patients for quality of life and functional status 6 months after intensive care unit admission. MEASUREMENTS AND MAIN RESULTS: Data regarding 521 patients including 1,932 daily judgments by nurses and doctors were analyzed. Disagreement on at least one of the daily judgments by nurses and doctors was found in 21% of all patients and in 63% of the dying patients. The disagreements more frequently concerned quality of life than survival. The higher the Simplified Acute Physiology Score and the longer the intensive care unit stay, the more divergent judgments were observed (p <.001). In surviving and dying patients, nurses gave more pessimistic judgment and considered withdrawal more often than did doctors (p <.001). Patients only rarely indicated bad quality of life (6%) and severe physical disability (2%) 6 months after intensive care unit admission. Compared with patients' own assessment, neither nurses nor doctors correctly predicted quality of life; false pessimistic and false optimistic appreciation was given. CONCLUSIONS: Disagreement between nurses and doctors was frequent with respect to their judgment of futility of medical interventions. Disagreements most often concerned the most severely ill patients. Nurses, being more pessimistic in general, were more often correct than doctors in the judgment of dying patients but proposed treatment withdrawal in some very sick patients who survived. Future quality of life cannot reliably be predicted either by doctors or by nurses. PMID- 12576950 TI - Cost of acute renal failure requiring dialysis in the intensive care unit: clinical and resource implications of renal recovery. AB - OBJECTIVE: Acute renal failure can be treated with continuous renal replacement therapy (CRRT) or intermittent hemodialysis. There is no difference in mortality, although patients treated with CRRT may have a higher rate of renal recovery. Given these considerations, an estimate of the costs by modality may help in choosing the method of dialysis. As such, the objective of this study was to estimate the cost of CRRT and intermittent hemodialysis in the intensive care unit and to explore the impact of renal recovery on subsequent clinical outcomes and costs among survivors. DESIGN: Retrospective cohort study of all patients who developed acute renal failure and required dialysis between April 1, 1996, and March 31, 1999. SETTING: Two tertiary care intensive care units in Calgary, Canada. PATIENTS: A total of 261 critically ill patients. INTERVENTIONS: None. MEASUREMENTS: All patients were followed to determine in-hospital and subsequent clinical outcomes (survival and frequency of renal recovery). The immediate and potential long-term costs of CRRT and intermittent hemodialysis were measured. MAIN RESULTS: The cost of performing CRRT ranged from Can 3,486 dollars to Can 5,117 dollars per week, depending on the modality and the anticoagulant used, and it was significantly more expensive than intermittent hemodialysis (Can 1,342 dollars per week). Survivors with renal recovery spent significantly fewer days in hospital (11.3 vs. 22.5 days, p<.001) and incurred less healthcare costs (11,192 dollars vs. 73,273 dollars, p<.001) over the year after hospital discharge compared with survivors who remained on dialysis. CONCLUSIONS: Immediate cost savings could be achieved by increasing the use of intermittent hemodialysis rather than CRRT for patients with acute renal failure in the intensive care unit. Because of the high cost of ongoing dialysis, CRRT may still be an economically efficient treatment if it improves renal recovery among survivors; further study in this area is required. PMID- 12576952 TI - Empirical antimicrobial therapy of septic shock patients: adequacy and impact on the outcome. AB - OBJECTIVE: To assess the adequacy of empirical antimicrobial therapy prescribed in septic shock patients and to evaluate the relationship between inadequate antimicrobial therapy and 30-day mortality. DESIGN: Prospective observational study. SETTING: Medical-surgical (16-bed) intensive care unit in an urban teaching hospital. PATIENTS: A total of 107 patients requiring intensive care admission were prospectively evaluated during the 3-yr period of the study. INTERVENTIONS: Prospective patient surveillance and data collection and assessment of antimicrobial therapy according to microbiological documentation. MEASUREMENTS AND MAIN RESULTS: A source of infection associated with a microbiological documentation was identified in 78 of the 107 patients (72%). Empirical antimicrobial therapy consisted of a pivotal antibiotic (beta-lactam) associated with an aminoglycoside (59 patients) or a fluoroquinolone (21 patients). Vancomycin was added in 14 patients. Sixty-nine of the 78 patients (89%) received an adequate antimicrobial therapy. The mortality rate of patients receiving an adequate antimicrobial therapy was 56%, and seven of the nine patients (78%) receiving an inadequate antimicrobial therapy died (p =.2). Among the 81 patients who were alive on day 3, antimicrobial therapy was modified in agreement to clinical status and microbiological documentation in 80% of cases, with de-escalation in 64% of cases. De-escalation consisted of withdrawing the nonpivotal antibiotic in 42% of patients or switching to a narrow-spectrum beta lactam antibiotic (22% of cases). CONCLUSION: The prescription of empirical antimicrobial therapy by a senior physician in agreement with practice guidelines made it possible to achieve a crude rate of 89% of adequate antimicrobial therapy in study patients. Inadequate antimicrobial therapy was associated with a 39% excess of mortality. A de-escalation of the empirical therapy was possible in 64% of patients. PMID- 12576953 TI - Controlled trial of oronasal versus nasal mask ventilation in the treatment of acute respiratory failure. AB - OBJECTIVE: Noninvasive positive pressure techniques such as continuous and bilevel positive airway pressure avoid intubation and its attendant complications in selected patients with acute respiratory failure. However, mask intolerance remains a common cause for failure of noninvasive ventilatory techniques. The aim of our study was to assess patient tolerance of oronasal vs. nasal mask ventilation in acute respiratory failure. DESIGN: Randomized, controlled trial. SETTING: Emergency department or intensive care units at a university hospital. PATIENTS: Seventy patients with acute respiratory failure as evidenced by clinical or blood gas criteria. INTERVENTIONS: Patients randomly received either a disposable nasal or an oronasal mask (Respironics, Pittsburgh, PA) when they met study criteria. MEASUREMENTS AND MAIN RESULTS: Thirty-five patients were randomized into each arm of the study; most of the patients had acute cardiogenic pulmonary edema (48.6% of the nasal mask group and 42.8% of the facial mask group) or chronic obstructive airway disease (34.3% of the nasal mask group and 31.4% of the facial mask group). Baseline clinical characteristics of the two groups of patients were similar. Heart and respiratory rates and blood gases improved similarly for patients in both mask groups. Rates of intubation were also similar (eight in each group). However, mask intolerance was significantly higher in the nasal than the oronasal mask group (12 vs. 4, respectively, p=.023). Four patients in the nasal (11.4%) and two in the oronasal mask group (5.7%) died later during the hospitalization. The overall success rate tended to be greater in the oronasal (65.7%) than the nasal group (48.6%), but the difference was not statistically significant. CONCLUSION: Although both masks performed similarly with regard to improving vital signs and gas exchange and avoiding intubation, the nasal mask was less well tolerated than the oronasal mask in patients with acute respiratory failure. PMID- 12576954 TI - Gastric capnometry with air-automated tonometry predicts outcome in critically ill patients. AB - CONTEXT: Contrary to tonometer gastric intramucosal pH, there is currently no validated threshold prognostic value for Pco2 gap (tonometer gastric mucosal Pco2 minus arterial Pco2) in the critically ill patient. OBJECTIVE: To demonstrate a relationship between Pco2 gap and mortality in mechanically ventilated patients. DESIGN AND SETTING: Inception cohort study from a 9-month prospective survey of 95 consecutively ventilated critically ill patients in a teaching hospital. PATIENTS: All the ventilated patients of the intensive care unit were included at their admission. MEASUREMENTS AND MAIN RESULTS: Gastric Pco2 using regional capnometry with air-automated tonometry, arterial gas, lactate, and organ system failure score were measured at admission and after 6, 12, 24, 48, 72, 96, and 120 hrs. For the entire population, the 28-day mortality was 44%. In multivariate analysis, independent predictors of death were organ system failure score (odds ratio, 2.12; 95% confidence interval, 1.02-3.14), 24-hr Pco2 gap (odds ratio, 1.57; 95% confidence interval, 1.10-2.24), and 24-hr lactate (odds ratio, 1.48; 95% confidence interval, 1.06-2.05). We found a threshold value of 20 mm Hg for Pco2 gap and 2.5 mmol/L for lactate, which was associated with a sensitivity of 0.70 and 0.72, respectively, and a specificity of 0.72 and 0.73, respectively. CONCLUSION: The Pco2 gap is a marker of mortality in ventilated patients in the intensive care unit. PMID- 12576955 TI - Use of ultrasound guidance in the insertion of radial artery catheters. AB - OBJECTIVE: To assess the role of a portable ultrasound device in the insertion of radial artery catheters. DESIGN: Prospective, randomized, comparative study. SETTING: Tertiary university hospital. PATIENTS: Elective surgery patients requiring arterial catheter insertion for intraoperative monitoring. INTERVENTIONS: A portable ultrasound device was used to visualize the radial artery at the wrist and to direct arterial catheter insertion. This new technique of arterial catheter insertion was compared with the classic palpation technique. MEASUREMENTS AND MAIN RESULTS: A total of 69 patients requiring an arterial catheter were randomized to either the ultrasound (34 patients) or palpation technique (35 patients). The time taken from skin puncture to successful arterial catheter insertion, the time taken per insertion attempt, the number of attempts required, and the number of cannulae used were recorded for each group. The arterial cannula was inserted on the first attempt in 21 (62%) cases using ultrasound vs. 12 (34%) cases by palpation (p =.03). Significantly fewer attempts were required for catheter insertion using ultrasound as compared with palpation (mean +/- sd, 1.6 +/- 1.0 vs. 3.1 +/- 2.4; p=.003); however, the time taken for each successful attempt was longer (26.1 +/- 2.0 vs. 17.3 +/- 1.6 secs, p=.001). A trend toward shorter overall time required for catheter insertion was found for the ultrasound group (55.5 +/- 63.8 vs. 111.5 +/- 121.5 secs, p=.17). There were four failures in the ultrasound group and one in the palpation group (not significant). CONCLUSIONS: Ultrasound is a useful adjunct to arterial catheter insertion and increases the rate of success at first attempt. The technique is easy to learn and may reduce the time taken to insert the catheter. PMID- 12576956 TI - Intravenous nicorandil prevents thiamylal-fentanyl-induced bronchoconstriction in humans. AB - OBJECTIVE: Nicorandil has a hybrid property between nitrates and potassium channel openers and has been reported to cause a concentration-dependent relaxation of isolated guinea pig trachealis. Experimental asthma in a guinea pig model was also inhibited by nicorandil. However, no clinical data on the bronchorelaxant effects of this drug have been published. The aim of this study was to investigate whether intravenous nicorandil prevents thiamylal-fentanyl induced bronchoconstriction. DESIGN: Double-blind, prospective, placebo controlled, randomized study. PATIENTS: A total of 36 patients were randomly allocated to two groups: a control group (n = 18) and a nicorandil group (n = 18). INTERVENTIONS: Intravenous administration of nicorandil or a placebo (normal saline). MEASUREMENTS AND MAIN RESULTS: Anesthesia was induced with 5 mg/kg thiamylal and 0.3 mg/kg vecuronium. A continuous infusion of 15 mg x kg(-1) x hr( 1) thiamylal was then used to maintain the anesthesia. Controlled ventilation was maintained, delivering 50% nitrous oxide in oxygen. At 20 mins after the induction of anesthesia, the control group patients were given a 6 mL/hr continuous infusion of normal saline and the nicorandil group patients were given a 6 mg x hr(-1) (6 mL/hr) continuous infusion of nicorandil for 60 mins. At 30 mins after the start of the study, both groups received a 5-microg/kg dose of fentanyl. Systolic and diastolic arterial pressure, heart rate, mean airway resistance, expiratory airway resistance, and dynamic lung compliance were measured for the baseline condition, just before the administration of fentanyl (T30), at three consecutive 6-min intervals after the fentanyl injection (T36, T42, and T48) and 30 mins after the fentanyl injection (T60). Both groups had comparable baseline values for mean airway resistance, expiratory airway resistance, and dynamic lung compliance. In the control group, both mean airway resistance and expiratory airway resistance increased significantly at T36-60, compared with the baseline values, and dynamic lung compliance decreased significantly at T36-60, compared with the baseline value. In the nicorandil group, no changes in mean airway resistance, expiratory airway resistance, or dynamic lung compliance occurred at T36-60. CONCLUSIONS: Our observations suggest that the intravenous administration of nicorandil has a bronchodilator effect in humans. PMID- 12576957 TI - Enteral nutrition with eicosapentaenoic acid, gamma-linolenic acid, and antioxidants reduces alveolar inflammatory mediators and protein influx in patients with acute respiratory distress syndrome. AB - OBJECTIVE: Previously, we showed that acute respiratory distress syndrome patients fed an enteral diet containing eicosapentaenoic acid and gamma-linolenic acid and elevated antioxidants (EPA+GLA; Oxepa) had significantly reduced pulmonary inflammation, increased oxygenation, and improved clinical outcomes. In a subset of acute respiratory distress syndrome patients from this trial, we performed a preliminary examination of the potential mechanisms underlying these clinical improvements by retrospectively testing the hypothesis that enteral feeding with EPA+GLA could reduce alveolar-capillary membrane protein permeability and the production of interleukin (IL)-8, IL-6, tumor necrosis factor-alpha, and leukotriene B4 that are responsible, in part, for pulmonary inflammation. DESIGN: Prospective, randomized, double-blind, controlled clinical trial. SETTING: Intensive Care Unit of the Ohio State University Medical Center. PATIENTS: A total of 67 patients were enrolled who met defined criteria for acute lung injury/acute respiratory distress syndrome. INTERVENTIONS: A total of 43 of 67 evaluable patients randomly received either EPA+GLA or an isonitrogenous, isocaloric standard diet that was tube fed at a minimum caloric delivery of 75% of basal energy expenditure times 1.33 for at least 4 to 7 days. MEASUREMENTS AND MAIN RESULTS: Bronchoalveolar lavage (BAL) was performed at baseline and study days 4 and 7 to obtain BAL fluid (BALF) for measurement of total protein, ceruloplasmin, and transferrin, total neutrophil count, IL-8, IL-6, tumor necrosis factor-alpha, and leukotriene B4. Oxygenation, measured as Pao2/Fio2, was assessed before BAL. Patients fed EPA+GLA had a significant reduction in BALF ceruloplasmin and IL-8 during the study as compared with patients fed the control diet. BALF levels of total protein, neutrophils, and leukotriene B4 tended to decrease in EPA+GLA patients over the course of the study as compared with control patients. BALF levels of IL-6 declined similarly during the study in both groups. A trend toward a reduction in BALF tumor necrosis factor-alpha was observed on study day 7 in the EPA+GLA group as compared with control patients. Significant improvements in oxygenation (Pao2/Fio2) occurred in EPA+GLA patients on study day 4 as compared with controls. Correlation analysis revealed significant relationships between BALF neutrophil counts and indices of alveolar capillary membrane protein permeability, IL-8, and leukotriene B4. CONCLUSIONS: This preliminary investigation showing a decrease in BALF levels of IL-8 and leukotriene B4 and the associated reduction of BALF neutrophils and alveolar membrane protein permeability in acute respiratory distress syndrome patients fed EPA+GLA support, in part, the potential mechanisms underlying the previously described clinical improvements with this diet. Additional controlled studies are needed to confirm these findings. PMID- 12576958 TI - Comparative virulence of Candida albicans yeast and filamentous forms in orally and intravenously inoculated mice. AB - OBJECTIVE: Candida albicans, a dimorphic fungus that switches from yeast to filamentous forms, is a major cause of complicating systemic infection in intensive care patients. The aim of this study was to compare the pathogenic potential of C. albicans yeast and filamentous forms. DESIGN: Separate groups of mice were inoculated either intravenously or orally with C. albicans CAF2 (wild type), HLC54 (yeast forms defective in filament formation), or BCa2-10 (constitutively filamentous). Mice were killed 1, 7, 14, and 21 days after intravenous C. albicans and kidneys and liver were quantitatively cultured; cohort groups were observed for mortality. Mice were pretreated with antibiotics for 3 days before oral inoculation with C. albicans, and killed 3 days later with dexamethasone administered for the latter 3 days; at sacrifice, the mesenteric lymph nodes and kidneys were cultured to monitor extraintestinal dissemination of C. albicans. SETTING: University teaching hospital research laboratory. SUBJECTS: Female, Swiss Webster, adult mice. MEASUREMENTS AND MAIN RESULTS: In intravenously inoculated mice, mortality was highest with wild-type C. albicans CAF2 (92%), intermediate with HLC54 (56%), and not detected with constitutively filamentous BCa2-10 (0%); BCa2-10 was cleared from the kidney and liver, but CAF2 and HLC54 were recovered at approximately 10(5-7)/g kidney and 10(4-5)/g liver. There was only occasional mortality in orally inoculated mice and the numbers of cecal C. albicans CAF2 and HLC54 were similarly high (approximately 10(7)/g), whereas numbers of cecal BCa2-10 were at least 100-fold lower. Extraintestinal dissemination was greatest with HLC54, intermediate with CAF2, and undetectable with BCa2-10. CONCLUSIONS: Of the three C. albicans strains studied, wild-type CAF2 was most virulent in intravenously inoculated mice and HLC54 (defective in filament formation) was most virulent in orally inoculated mice. The constitutively filamentous BCa2-10 was avirulent in both models, suggesting that filamentous forms by themselves might not be critically important for C. albicans virulence. PMID- 12576959 TI - Effect of endogenous nitric oxide on hyperoxia and tumor necrosis factor-alpha induced leukosequestration and proinflammatory cytokine release in rat airways. AB - OBJECTIVE: To investigate the effects of endogenous nitric oxide on hyperoxia and tumor necrosis factor-alpha-induced leukosequestration and proinflammatory cytokine release in rat airways. DESIGN: Prospective, randomized, controlled animal study. SETTING: Experimental laboratory. SUBJECTS: Male Sprague-Dawley rats weighing 350-500 g. INTERVENTIONS: The rats were pretreated with N(G)-nitro L-arginine methyl ester (L-NAME; 10 mg/kg) or saline intravenously 4-6 mins before intratracheal administration of tumor necrosis factor-alpha, 95% oxygen, or both, when the vasopressor effect of L-NAME had reached a plateau. MEASUREMENTS AND MAIN RESULTS: Bronchoalveolar lavage fluid was recovered from the airway of rats after exposure to 95% oxygen and tumor necrosis factor-alpha for 6 hrs under ventilator support. Neutrophils in lavage fluid were isolated and examined for the inducible nitric oxide synthase expression by flow-cytometric assay. Tumor necrosis factor-alpha and interleukin-1 beta in lavage fluid were measured by enzyme-linked immunosorbent assay. The percentage of neutrophils in bronchoalveolar fluid was significantly higher in rats exposed to hyperoxia + tumor necrosis factor-alpha (29.7 +/- 12.5%) compared with rats with hyperoxia (16.3 +/- 1.2%), tumor necrosis factor-alpha (4.2 +/- 1.1%), or room air (5.0 +/- 1.8%) alone (p <.05). Rats exposed to hyperoxia + tumor necrosis factor-alpha had significantly higher concentrations of inducible nitric oxide synthase of neutrophils (350.1 +/- 75.7 mean fluorescence intensity), compared with rats with hyperoxia (64.9 +/- 1.6 mean fluorescence intensity), tumor necrosis factor-alpha (102.6 +/- 15.3 mean fluorescence intensity), or room air (111.2 +/- 25.8 mean fluorescence intensity) alone (p <.05). Rats exposed to hyperoxia + tumor necrosis factor-alpha significantly produced higher concentrations of tumor necrosis factor-alpha and interleukin-1 beta, compared with rats with tumor necrosis factor-alpha, hyperoxia, or room air alone. Hyperoxia + tumor necrosis factor-alpha also significantly increased growth-related oncogene/cytokine induced neutrophil chemoattractant (GRO/CINC)-1 in bronchoalveolar fluid, compared with those receiving tumor necrosis factor-alpha alone, hyperoxia alone, or room air alone. L-NAME significantly enhanced the percentage of neutrophil recovery and the production of tumor necrosis factor-alpha, interleukin-1 beta, and GRO/CINC-1 in airways compared with the corresponding hyperoxia + tumor necrosis factor-alpha treatment alone. CONCLUSIONS: Endogenous nitric oxide may be an important endogenous inhibitor of hyperoxia + tumor necrosis factor-alpha induced leukocyte recruitment and subsequently tumor necrosis factor-alpha, interleukin-1 beta, and GRO/CINC-1 release. PMID- 12576960 TI - Time course for inhibition of lipopolysaccharide-induced lung injury by genistein: relationship to alteration in nuclear factor-kappaB activity and inflammatory agents. AB - OBJECTIVE: This study determined the time course for inhibition of lipopolysaccharide-induced acute lung injury following a single dose of genistein. In addition, the study investigated whether a multiple dosing schedule with genistein retained the inhibitory effects on acute lung injury, nuclear factor-kappaB activation, and production of nuclear factor-kappaB-dependent inflammatory agents, such as matrix metalloproteinase-9 and nitric oxide. DESIGN: Prospective, randomized, laboratory study. SETTING: Experimental laboratory at a university. SUBJECTS: Rats weighing 280-300 g. INTERVENTIONS: Saline or lipopolysaccharide (6 mg/kg of body weight) administered intratracheally with a single dose of genistein (50 mg/kg) or a multiple dosing schedule with genistein (16 mg/kg every 6 hrs for 2 days with lipopolysaccharide treatment at 24 hrs after the first administration of genistein). MEASUREMENTS AND MAIN RESULTS: A 2 hr pretreatment with genistein (a single dose) inhibited biochemical lung injury variables as well as neutrophil infiltration with a maximal inhibition at 4 hrs after lipopolysaccharide treatment. These inhibitory effects of genistein declined with time and were no longer significant by 14-24 hrs after lipopolysaccharide treatment. The multiple dosing schedule with genistein retained significant inhibitory effects on biochemical lung injury variables and the number of neutrophils in the bronchoalveolar lavage fluid at 24 hrs after lipopolysaccharide treatment compared with a single pretreatment with genistein. The multiple dosing schedule with genistein also enhanced the inhibition of induced nuclear factor-kappaB activity as well as matrix metalloproteinase-9 activity and nitric oxide production at 24 hrs after lipopolysaccharide treatment. CONCLUSIONS: This study reports the time course of the inhibitory effects of a single genistein pretreatment on acute lung injury with the maximal effects at 4 hrs after lipopolysaccharide treatment. However, a multiple dosing schedule with genistein retained the inhibitory effect on acute lung injury at 24 hrs after lipopolysaccharide treatment. The mechanisms by which genistein exerts an inhibitory effect on acute lung injury may involve the suppression of nuclear factor-kappaB activation, matrix metalloproteinase-9 activity, and NO production. PMID- 12576961 TI - Heparin mobilizes xanthine oxidase and induces lung inflammation in acute pancreatitis. AB - OBJECTIVE: To evaluate the effect of low molecular weight heparin on plasma xanthine oxidase concentrations and lung inflammatory response during acute pancreatitis. DESIGN: Randomized, controlled trial. SETTING: Experimental laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Acute pancreatitis was induced by intraductal administration of 5% sodium taurocholate. Low molecular weight heparin (0, 30, 90, or 300 units/kg) was administered immediately after induction of pancreatitis. MEASUREMENTS AND MAIN RESULTS: Lipase and xanthine oxidase plasma concentrations were measured 3 hrs after pancreatitis induction. Expression of P-selectin messenger RNA and myeloperoxidase activity as a marker of neutrophil infiltration were determined in the lung. An increase in xanthine oxidase plasma concentrations was observed during pancreatitis. Administration of heparin also increased plasma xanthine oxidase activity in both control and pancreatitis animals. Measures of xanthine oxidase present in the endothelial surface indicate that during pancreatitis, the enzyme is released from the gastrointestinal endothelium. By contrast, heparin mobilizes xanthine oxidase from almost all organs evaluated. Neutrophil infiltration was increased in the lung during pancreatitis. Heparin administration further increased, in a dose dependent manner, myeloperoxidase activity and P-selectin expression in the lung in animals with pancreatitis. By contrast, in control animals, heparin had no effect on myeloperoxidase activity and did not induce P-selectin up-regulation. CONCLUSION: During acute pancreatitis, heparin administration might mobilize xanthine oxidase attached to endothelial cells, originating a free radical generating system in the circulation that would trigger an inflammatory response in the lung. PMID- 12576962 TI - Induced hyperthermia exacerbates neurologic neuronal histologic damage after asphyxial cardiac arrest in rats. AB - BACKGROUND: Temperature is an important modulator of the evolution of ischemic brain injury--with hypothermia lessening and hyperthermia exacerbating damage. We recently reported that children resuscitated from predominantly asphyxial arrest often develop an initial spontaneous hypothermia followed by delayed hyperthermia. The initial hypothermia observed in these children was frequently treated with warming lights which, despite careful monitoring, often resulted in overshoot hyperthermia. We have previously reported in a rat model of asphyxial cardiac arrest that active warming, to prevent spontaneous hypothermia, worsens brain injury. OBJECTIVE: We sought to determine whether delayed induction of hyperthermia would worsen brain injury after asphyxial arrest in rats. DESIGN: Male Sprague-Dawley rats were asphyxiated for 8 mins and resuscitated. An implantable temperature probe was placed into the peritoneum before asphyxia. The probe is a component of a computer-based, radiofrequency, telemetry system (Minimitter, Sunriver, OR) that allowed continuous acquisition and manipulation (via heating and cooling devices) of core (intraperitoneal) body temperature. Body temperature was monitored but not manipulated for the first 24 hrs of recovery. Rats were assigned to: no temperature manipulation (n = 21), induced hyperthermia (40 +/- 0.5 degrees C) for 3 hrs beginning at 24 hrs (n = 21), or induced hyperthermia at 48 hrs (n = 10). Control groups included sham rats (all surgical procedures except asphyxia) treated with induced hyperthermia at 24 hrs (n = 4) or 48 hrs (n = 4) and naive rats (n = 4). Rats were killed at 7 days and injured neurons in hematoxylin and eosin stained coronal brain sections through dorsal hippocampus were scored in a semiquantitative manner on a scale of 0 to 10 (0 = normal; 1 = up to 10% neurons with ischemic neuronal changes; 10 = 90-100% neurons with ischemic neuronal changes). Normal-appearing neurons were also counted in CA1. The number of normal-appearing neurons in a 20x field in CA1 were also counted. MAIN RESULTS: All naive and sham hyperthermia control rats survived the protocol. There was a trend toward a larger mortality rate in asphyxiated rats treated with induced hyperthermia at 24 hrs (9 of 21 died) vs. asphyxiated rats without induced hyperthermia (3 of 21) or with hyperthermia induced at 48 hrs (3 of 10) (Kaplan-Meier p=.0595). Asphyxiated rats with hyperthermia induced at 24 hrs had larger (worse) histopathology damage scores than rats subjected to asphyxia without induced hyperthermia (9.3 +/- 1.5 vs. 6.2 +/- 2.6; p=.001). Histopathology damage scores in asphyxiated rats with hyperthermia induced at 48 hrs did not differ from those in rats asphyxiated without induced hyperthermia (6.4 +/- 3.0 vs. 6.2 +/- 2.6; p=.907). There were fewer normal-appearing CA1 neurons in asphyxiated rats with hyperthermia induced at 24 hrs vs. rats subjected to asphyxia without induced hyperthermia (33 +/- 13 vs. 67 +/- 36; p=.002). The number of normal-appearing CA1 neurons in asphyxiated rats with hyperthermia induced at 48 hrs did not differ from that in rats asphyxiated without induced hyperthermia (59 +/- 21 vs. 67 +/- 36; p=.885). CONCLUSIONS: Induced hyperthermia when administered at 24 hrs, but not 48 hrs, worsens ischemic brain injury in rats resuscitated from asphyxial cardiac arrest. This may have implications for postresuscitative management of children and adults resuscitated from cardiac arrest. The common clinical practice of actively warming patients with spontaneous hypothermia might result in iatrogenic injury if warming results in hyperthermic overshoot. Avoidance of hyperthermia induced by active warming at critical time periods after cardiac arrest may be important. PMID- 12576963 TI - Imbalance between matrix metalloproteinases (MMP-9 and MMP-2) and tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) in acute respiratory distress syndrome patients. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) are known to be involved in degradation of extracellular matrix. We aimed to assess the role of MMPs and their natural inhibitors (TIMPs) in the genesis and the evolution of acute respiratory distress syndrome (ARDS). DESIGN: Prospective, clinical study. SETTING: Intensive care unit of a university hospital. PATIENTS: Twenty-one patients were assigned to three different groups: Group 1 patients developed ARDS that rapidly resolved in <4 days; Group 2 patients developed ARDS lasting >8 days; Group 3 (control group) patients had clinical criteria for hospital acquired pneumonia without ARDS. INTERVENTION: Bronchoalveolar lavages were performed on day 0 of the onset of ARDS and on days 4, 8, and 12 for unresolving ARDS. For group 3, the bronchoalveolar lavages were performed on day 0 of the pneumonia. On these bronchoalveolar lavage fluids, we measured the amount of MMP 9 and -2 and their inhibitors TIMP-1 and -2. MEASUREMENTS AND MAIN RESULTS: The amount of MMP-9 measured by enzyme-linked immunosorbent assay was significantly lower in the bronchoalveolar lavages from patients with ARDS (group 1 and group 2) compared with the control group (p <.01) throughout the study. The ratio MMP 9/TIMP-1 was also significantly smaller and was less than one in the two ARDS groups (p <.05) compared with the control group (group 3), where this ratio was greater than one. In the second bronchoalveolar lavages, this ratio was greater than one only in the ARDS group that rapidly resolved (group 1), whereas it stayed less than one when the ARDS was lasting (group 2). Concerning the quantity of MMP-2 and the ratio MMP-2/TIMP-2, there was no statistical difference between the three groups throughout the study. Using zymography, there was no significant difference in the amounts of active and latent MMP-9 between the three groups. Moreover, no significant difference in the quantity of latent and active MMP-2 in the three groups was noted. CONCLUSION: These results suggest that the MMP-9 level and MMP-9/TIMP-1 ratio play a role in the pathogenesis of ARDS and, namely, the imbalance between MMP-9 and TIMP-1 would participate in airway remodeling leading to either short- or long-course ARDS. The ratio MMP-9/TIMP-1 could be a predictive factor of the ARDS evolution. PMID- 12576964 TI - Performance of the partial CO2 rebreathing technique under different hemodynamic and ventilation/perfusion matching conditions. AB - OBJECTIVE: The partial CO2 rebreathing technique has been demonstrated to accurately measure the effective pulmonary capillary blood flow (PCBF) in different clinical situations. Usually, PCBF is calculated from changes in CO2 elimination (VCO2) and end-tidal partial pressure of CO2 (PetCO2 ), which can be obtained noninvasively. In this study, we investigated the performance of the partial CO2 rebreathing technique under different conditions of ventilation/perfusion matching and hemodynamic states. In addition, we investigated whether the determination of arterial blood gases combined with mathematical modeling of gas exchange can improve the performance of this method. DESIGN: Prospective, controlled animal laboratory study. SETTING: Experimental research facility of a university hospital. SUBJECTS: Sixteen female sheep weighing 45-55 kg. INTERVENTIONS: Cardiac output and ventilation/perfusion matching were manipulated during three phases: phase I, variation in cardiac output to achieve normal, hyperdynamic and hypodynamic states; phase II, increase of alveolar deadspace and variation in cardiac output; phase III, lung injury and increased alveolar deadspace. Partial CO2 rebreathing maneuvers were performed to obtain variations in VCO2 and PetCO2 between a nonrebreathing (NR) and a rebreathing (R) period. MEASUREMENTS AND MAIN RESULTS: PCBF was measured by the rebreathing method as PCBF = -DeltaVCO2/f(Pc'CO2 (R), Pc'CO2(NR), Hb), where f is the CO2 dissociation curve in blood, Pc'CO2 is the end-capillary partial pressure of CO2, Delta is the variation between NR and R periods, and Hb is hemoglobin concentration. Pc'CO2 was estimated from PetCO2 according to two algorithms. In the so-called "noninvasive algorithm," Pc'CO2 = PetCO2, with PetCO2(NR) and PetCO2(R) being determined as the mean PetCO2 value of the last 60 secs preceding rebreathing and within 15-30 secs of rebreathing, respectively. In the "semi invasive algorithm," Pc'CO2(NR) was estimated as the PaCO2, and Pc'CO2(R) was estimated as follows: First, a monoexponential function was fitted to PetCO2 values during rebreathing and the asymptote represented PetCO2(R). Second, the Pc'CO2(R) to PetCO2(R) difference was calculated by means of a bicompartmental, tidal model of gas exchange, which showed that such differences decrease with the degree of rebreathing. PCBF values obtained with both algorithms were compared with thermodilution cardiac output minus intrapulmonary shunt flow. Bias and precision calculations with the noninvasive algorithm in phases I, II, and III were, respectively, -1.0 +/- 1.9, -2.1 +/- 2.6, and -2.4 +/- 1.2 L/min. The semi invasive algorithm had an overall better performance in the phases investigated: 1.2 +/- 1.9, -0.6 +/- 2.0, and -0.2 +/- 3.0 L/min, respectively. The noninvasive algorithm showed a slight tendency to overestimate lower reference PCBF values and, importantly, to underestimate higher PCBF values in all three phases (r = .66, p<.0001; r = -.75, p<.001; r = -.60, p<.0001, respectively). A similar figure was observed with the semi-invasive algorithm in phase I (r = -.47, p<.01) but not in phases II and III (r = -.1, p=.54; r =.62, p<.001, respectively). CONCLUSIONS: Although PCBF is systematically underestimated during hyperdynamic cardiac output states and high alveolar deadspaces, the performance of the partial CO2 rebreathing technique can be improved by means of arterial blood gas sampling and an algorithm that takes in account the effects of nonequilibration of PetCO2 during rebreathing and the variation of Pc'CO2 to PetCO2 differences from the nonrebreathing to the rebreathing period. Such an algorithm may prove useful under moderately increased alveolar deadspace and normal to hypodynamic cardiac output states. PMID- 12576965 TI - Noninvasive mechanical ventilation in clinical practice: a 2-year experience in a medical intensive care unit. AB - OBJECTIVE: To evaluate the feasibility and outcome results of noninvasive mechanical ventilation (NIV) in daily clinical practice outside any prospective protocol-driven trial. DESIGN: An observational retrospective cohort study. SETTING: A 22-bed medical intensive care unit in a university hospital. PATIENTS: A consecutive cohort of 124 patients who underwent 143 NIV trials, regardless of the indication, over two consecutive years (1997-1998). INTERVENTIONS: None. RESULTS: A total of 604 acute respiratory failure patients underwent mechanical ventilation, and 143 NIVs were performed in 124 patients. The overall prevalence of NIV use was 143 of 604 patients (24%) in three groups: hypoxemic acute respiratory failure (29.5%), hypercapnic acute respiratory failure (41%), and weaning/postextubation (29.5%). Intubation was avoided in 92 of 143 of the NIVs performed (64%), 19 (13%) after changing the initial NIV mode (i.e., a success rate of 62%, 51%, and 86% in the three groups, respectively). A total of 35 of 51 intubated patients (69%) required intubation during the first 24 hrs of NIV. Intensive care unit stay was 12 +/- 10 days for the overall population, and mortality, when NIV failed, was 13 of 124 patients (10.5%). Arterial pH (p =.0527) and the Pao2/Fio2 ratio (p =.0482) after 1 hr were the only independent predictive factors for NIV failure by multivariate analysis. CONCLUSIONS: This study confirms the results of controlled trials and demonstrates the feasibility and efficacy of NIV applied in daily clinical practice. These results suggest that NIV should be considered as a first-line ventilatory treatment in various etiologies of acute respiratory failure and as a promising weaning technique and postextubation ventilatory support. However, NIV should certainly be performed by a motivated and sufficiently trained care team. PMID- 12576966 TI - Endotoxin increases both protein and fluid microvascular permeability in cat skeletal muscle. AB - OBJECTIVE: To evaluate effects of lipopolysaccharide (endotoxin) on protein and fluid permeability in a whole organ skeletal muscle preparation. DESIGN: Controlled, prospective laboratory study. SETTING: University research laboratory. SUBJECTS: Eleven adult male cats. INTERVENTIONS: The study was performed on the autoperfused and denervated calf muscles of the cat hindlimb placed in a fluid-filled plethysmograph. The endotoxin-induced change in the osmotic reflection coefficient for albumin was used as a measure of alteration in protein permeability of the microvascular wall, and the simultaneous change in capillary filtration coefficient was used as a measure of alteration in fluid permeability. Endotoxin as a bolus infusion (1 mg/kg iv) was given to six cats, and another five cats given only the vehicle (NaCl) were used as control. MEASUREMENTS AND MAIN RESULTS: Arterial blood flow, arterial and venous blood pressures, total vascular resistance, and tissue volume changes were measured continuously. The ratio between the osmotic reflection coefficients for albumin on two occasions (before and about 1.5 hr after endotoxin infusion) was calculated from the Starling fluid equilibrium equation. This was performed by measurement of the maximum absorption rate from an isovolumetric state by an intravenous bolus infusion of 20% human albumin (0.6 g/kg) and the capillary filtration coefficient. Albumin concentrations were measured before and after the albumin infusion to correct for effects of difference in plasma volume on the induced increase in colloid osmotic pressure. We found that the osmotic reflection coefficient for albumin was reduced by 30% (p <.05), and the capillary filtration coefficient was increased by 31% (p <.05) by endotoxin. No changes were seen in the vehicle experiments. CONCLUSION: Endotoxin causes a significant increase in both protein and fluid microvascular wall permeability. These effects may explain the marked leakage of plasma to the interstitium that is often seen in critically ill patients with sepsis and systemic inflammatory response syndrome. PMID- 12576967 TI - Cytokines down-regulate alpha1-adrenergic receptor expression during endotoxemia. AB - OBJECTIVE: The reduced pressure response to norepinephrine in septic patients has directed our interest to the regulation of alpha1-adrenergic receptors in vitro and in vivo during conditions mimicking acute sepsis. DESIGN: Prospective animal trial followed by a controlled cell culture study. SETTING: Laboratory of the Department of Anesthesiology. SUBJECTS: Male Sprague-Dawley rats weighing 200 to 250 g and a mesangial cell line. INTERVENTIONS: Experimental endotoxemia was induced in rats with lipopolysaccharide, and blood pressure dose-response studies with norepinephrine were performed. Alpha1-receptor gene expression was determined in various organs by a specific RNase protection assay, and tissue concentrations of the proinflammatory cytokines interleukin-1beta and tumor necrosis factor-alpha were measured. Rat renal mesangial cells were incubated with these cytokines or with nitric oxide donors to investigate the regulation of alpha1-adrenergic receptors during severe inflammation on a cellular level. MEASUREMENTS AND MAIN RESULTS: The pressor effect of norepinephrine was markedly diminished during endotoxemia. The animals showed down-regulated mRNA levels of alpha1A-, alpha1B- and alpha1D-receptors in all organs investigated, and the tissue concentrations of interleukin-1beta and tumor necrosis factor-alpha were highly increased during experimental endotoxemia. Incubation of cultured rat renal mesangial cells with the cytokines resulted in diminished alpha -receptor gene expression and [3H]prazosin binding capacity, whereas incubation of the cells with nitric oxide donors did not affect alpha1B-receptor expression. In line, blocking of cytokine-induced nitric oxide synthesis by coincubation of mesangial cells with N(G)-nitro-L-arginine methyl ester did not influence cytokine-induced down-regulation of alpha1B-receptors. CONCLUSIONS: Our data show that endotoxemia causes a systemic down-regulation of alpha1-receptors on the level of gene expression and suggest that this effect is likely mediated by proinflammatory cytokines in a synergistic but nitric oxide-independent fashion. We propose that this down-regulation of alpha1-adrenergic receptors contributes to the attenuated blood pressure response to norepinephrine and, therefore, to septic circulatory failure in patients. PMID- 12576968 TI - Should vasopressin replace adrenaline for endotracheal drug administration? AB - OBJECTIVE: Arginine vasopressin was established recently as a drug of choice in the treatment of cardiac arrest and in retractable ventricular fibrillation; however, the hemodynamic effect of vasopressin following endotracheal drug administration has not been fully elucidated. We compared the effects of endotracheally administered vasopressin vs. adrenaline on hemodynamic variables in a canine model, and we investigated whether vasopressin produces the same deleterious immediate blood pressure decrease as did endotracheal adrenaline in the canine model. DESIGN: Prospective controlled study. SETTING: Animal laboratory in Tel-Aviv University, Israel. SUBJECTS: Five adult mongrel dogs weighing 6.5-20 kg. INTERVENTIONS: Dogs were anesthetized; each dog was intubated orally, and both femoral arteries were cannulated for the measurement of arterial pressure and for sampling blood gases. Each dog was studied four times, 1 wk apart, by using the same protocol for injection and anesthesia: endotracheal placebo (10 mL NaCl 0.9%,), endotracheal vasopressin (1 units/kg), endobronchial adrenaline (0.1 mg/kg), and endotracheal adrenaline (0.1 mg/kg). Following placebo, vasopressin, and adrenaline instillation, five forced manual ventilations were delivered with an Ambu bag. Each dog was its own control. MEASUREMENTS AND MAIN RESULTS: Following placebo or drug administration, heart electrocardiography and arterial pressures were continuously monitored with a polygraph recorder for 1 hr. Endotracheal vasopressin produced an immediate increase of diastolic blood pressure (from 83 +/- 10 mm Hg [baseline] to 110 +/- 5 mm Hg at 1 min postinjection). This response lasted >1 hr. In contrast, both endotracheal and endobronchial administration of adrenaline produced an early and significant (p <.05) decrease in diastolic and mean blood pressures. The diastolic blood pressure increase from 85 +/- 10 mm Hg to 110 +/- 10 mm Hg took an ill-afforded 55 secs following endotracheal adrenaline. Diastolic blood pressure was significantly (p <.05) higher following vasopressin compared with adrenaline administration in both routes. CONCLUSIONS: Vasopressin accomplishes its hemodynamic effect, particularly on diastolic blood pressure, more rapidly, vigorously, and protractedly and to a significant degree compared with both endotracheal and endobronchial adrenaline. Evaluation of the effects of endotracheal vasopressin in a closed chest cardiopulmonary resuscitation model is recommended. PMID- 12576969 TI - Recombinant antithrombin attenuates pulmonary inflammation following smoke inhalation and pneumonia in sheep. AB - OBJECTIVE: The interaction between coagulation and inflammation has become one of the major topics in critical care medicine. In the present study, we investigated the effect of posttreatment of sepsis with recombinant human antithrombin. DESIGN: Experimental laboratory in a university hospital. SETTING: University laboratory. SUBJECTS: Female merino ewes (n = 16). INTERVENTIONS: After 1 wk of recovery from the surgical preparation, a tracheotomy was performed followed by insufflation of 48 breaths of cotton smoke (<40 degrees C). Afterward, a stock solution of live (5 x 10(11) colony-forming units) was instilled in the both lung lobes through a bronchoscope. All sheep were mechanically ventilated employing 100% oxygen. An infusion of recombinant human antithrombin (100 units x kg(-1) x 24 hrs(-1), intravenously; n = 6) or saline (n = 6) was started 1 hr after injury. Sham control animals (n = 4) were surgically prepared but not insufflated with smoke and bacteria. Lung histologic changes were evaluated by a scoring system. MEASUREMENTS AND MAIN RESULTS: The infusion of recombinant human antithrombin maintained the baseline antithrombin activity throughout the study; in the saline-treated group, antithrombin activity decreased significantly. The lung wet/dry weight ratio and the histology score (combined scores for congestion, edema, inflammation, and hemorrhage) were significantly increased by the insult, but recombinant human antithrombin attenuated these responses. More than 30% of both bronchi and bronchioles were obstructed by cast formation after smoke inhalation and pneumonia. The cast was composed of epithelial cells, neutrophils, mucus, and fibrin. The obstruction was significantly improved by recombinant human antithrombin infusion. Arterial pressure and urine output were also attenuated in recombinant human antithrombin-treated animals. The increases in plasma nitrate/nitrite concentrations and pulmonary shunt fraction after the injury were not attenuated by recombinant human antithrombin. CONCLUSION: Posttreatment by recombinant human antithrombin was effective in treating acute lung injury after smoke inhalation and pneumonia in sheep. We hypothesize that the decrease in antithrombin activity during sepsis might induce severe airway obstruction and that supplementation with antithrombin inhibits this decrease. PMID- 12576970 TI - Increases in spinal fluid osmolarity induced by mannitol. AB - OBJECTIVE: Mannitol is widely used in hospitals worldwide to treat patients with high intracranial pressure and/or cerebral edema. One of the mechanisms by which mannitol is thought to affect intracranial pressure is by increasing the patient's serum osmolarity, but not the osmolarity in the brain or cerebrospinal fluid. In this way, mannitol is thought to increase the osmolarity gap between the brain and the blood, which in turn leads to removal of excess water from the brain. However, relatively little is known regarding long-term effects of mannitol on osmolarity of cerebrospinal fluid. We therefore sought to determine the effects of mannitol administration on the osmolarity of cerebrospinal fluid. DESIGN: Controlled trial. SETTING: University teaching hospital. PATIENTS: Patients with severe head injury and patients with subarachnoid bleeding who required insertion of an intracranial probe. MEASUREMENTS AND MAIN RESULTS: Serum and cerebrospinal fluid osmolarity were measured before and during mannitol administration in ten patients treated with mannitol for >or=72 hrs (group 1), ten patients treated for 24 to 48 hrs (group 2), and ten controls (group 3). Serum osmolarity increased quickly in all patients receiving mannitol (groups 1 and 2), whereas remaining constant in controls. Average cerebrospinal fluid osmolarity slowly increased in all patients receiving mannitol; cerebrospinal fluid osmolarity increased from (mean +/- sd) 291.5 +/- 4.0 to 315.5 +/- 4.5 mOsm/kg after 96 hrs in group 1 (p <.01), and from 288.9 +/- 3.5 to 296.9 +/- 6.2 mOsm/kg after 48 hrs in group 2 (p <.01). Cerebrospinal fluid osmolarity remained constant in controls (p <.01 for group 1 vs. group 3 and for group 2 vs. group 3, respectively). In group 1, the gap between serum and cerebrospinal fluid osmolarity initially increased (which was the desired effect), but later decreased first to baseline values and then to below-normal levels. CONCLUSIONS: Long-term administration of mannitol can induce significant increases in cerebrospinal fluid osmolarity in patients with subarachnoid hemorrhage or severe head injury. This may be an undesirable and potentially dangerous effect. Therefore, cerebrospinal fluid osmolarity should be measured regularly in all patients receiving mannitol for longer than 24 hrs. If cerebrospinal fluid osmolarity increases, discontinuation or tapering of mannitol therapy should be considered. PMID- 12576972 TI - Influence of the critically ill state on host-pathogen interactions within the intestine: gut-derived sepsis redefined. AB - Severe and prolonged states of catabolic stress have been shown to have profound effects on the intestinal tract microflora and intestinal function. Gut-derived sepsis is a term used to describe a state of systemic inflammation with organ dysfunction after severe catabolic stress hypothesized to be initiated and perpetuated by the intestinal tract microflora. Popular notions of the mechanism of this process have suggested that stress promotes the translocation of intestinal bacteria or their toxins into the systemic compartment resulting in the release of proinflammatory cytokines which participate in the systemic inflammatory response syndrome. This review is an attempt to redefine the mechanism of gut-derived sepsis by focusing on molecular events that result from host-pathogen interactions within the intestinal tract itself. This evidence based review posits that gut-derived bacteremia, even with potent nosocomial pathogens, is an event of low proinflammatory potential and, itself, is an insufficient stimulus for the systemic inflammatory response and organ failure state typically seen after severe and prolonged catabolic stress. Mechanisms of this apparent paradox are discussed. PMID- 12576971 TI - Arginine and nitric oxide metabolism in critically ill septic pediatric patients. AB - OBJECTIVE: To investigate whole body, arginine metabolism and nitric oxide synthesis rates in septic, critically ill pediatric patients. DESIGN: Prospective study. SETTING: Pediatric intensive care unit at a general hospital. PATIENTS: Ten consecutive septic patients age 6-16 yrs. INTERVENTIONS: Septic patients received an 8-hr primed, constant intravenous tracer infusion of L-[guanidino 15N2]arginine, L-[1-13C]leucine, and [13C]urea. A 24-hr urine collection was obtained for determination of [15N]nitrate enrichment (15NO3(-)) and urinary nitrogen. The next day they received an infusion of L-[5-13C]arginine and L-[5 13C-ureido, 5,5, 2H2]citrulline. Blood samples were obtained for determination of plasma isotopic enrichment of the tracers given and of derived [15N]citrulline (nitric oxide synthesis), L-[13C-guanidino 5,5, 2H2]arginine (M+3 arg) (arginine synthesis), and [15N]urea (urea formation). Data are compared with historic controls from studies in healthy young adults. MEASUREMENTS AND MAIN RESULTS: Plasma arginine fluxes were 67 +/- 21 and 72 +/- 17 micromol x kg(-1) x hr(-1), respectively, for the [15N2 guanidino] and the [13C] arginine labels, which were not different from reported adult values. The rates of arginine oxidation were 22.9 +/- 10.8 micromol x kg(-1) x hr(-1) and were higher than arginine synthesis rates of 9.6 +/- 4.2 micromol x kg(-1) x hr(-1) (p <.01); therefore, these patients were in a negative arginine balance. The rates of nitric oxide synthesis as estimated by the [15N]citrulline method were 1.58 +/- 0.69 micromol x kg(-1) x hr(-1) for septic patients and higher (p <.05) than values of 0.96 +/- 0.1 micromol x kg(-1) x hr(-1) in healthy adults. Septic patients were in a negative protein (leucine) balance of about -1.00 +/- 0.40 g x kg(-1) x day(-1). CONCLUSIONS: Homeostasis of plasma arginine in septic patients was impaired compared with reported adult values. The rates of arginine oxidation were increased whereas net arginine synthesis was unchanged, leading to a negative arginine balance. The rates of nitric oxide synthesis and the fraction of plasma arginine used for nitric oxide and urea formation were increased. These findings suggest that under condition of sepsis, arginine becomes essential in critically ill children. PMID- 12576973 TI - Appropriate use of antimicrobial agents: challenges and strategies for improvement. AB - The use of inadequate empirical antimicrobial therapy is common in intensive care unit patients and contributes to a number of poor outcomes. Selecting appropriate antimicrobial therapy is complicated by many factors, including the large number of agents available, the presence of resistant organisms, and the general desire among practitioners to use the most focused therapy available. An important aspect of appropriate antimicrobial use is prompt initiation of adequate empirical therapy, which has been shown to improve mortality rates in hospitalized patients with pneumonia and other serious infections. Other key strategies include streamlining antimicrobial therapy when a pathogen is identified and switching from intravenous to oral therapy when clinically indicated. In addition, antibiotic rotation (or cycling) has been evaluated in several trials as a means to minimize resistance. Promoting appropriate antimicrobial therapy ultimately will require a multidisciplinary, system oriented, institution-specific approach because each intensive care unit has its own unique flora and antimicrobial resistance patterns. PMID- 12576974 TI - Treatment of space-occupying cerebral infarction. AB - OBJECTIVE: Patients with a hemispheric infarct accompanied by massive edema have a poor prognosis; the case fatality rate may be as high as 80%, and most survivors are left severely disabled. Various treatment strategies have been proposed to limit brain tissue shifts and to reduce intracranial pressure, but their use is controversial. We performed a systematic search of the literature to review the evidence of efficacy of these therapeutic modalities. DATA SOURCES: Literature searches were carried out on MEDLINE and PubMed. STUDY SELECTION: Studies were included if they were published in English between 1966 and February 2002 and addressed the effect of osmotherapy, hyperventilation, barbiturates, steroids, hypothermia, or decompressive surgery in supratentorial infarction with edema in animals or humans. DATA SYNTHESIS: Animal studies of medical treatment strategies in focal cerebral ischemia produced conflicting results. If any, experimental support for these strategies is derived from studies with animal models of moderately severe focal ischemia instead of severe space-occupying infarction. None of the treatment options have improved outcome in randomized clinical trials. Two large nonrandomized studies of decompressive surgery yielded promising results in terms of reduction of mortality and improvement of functional outcome. CONCLUSIONS: There is no treatment modality of proven efficacy for patients with space-occupying hemispheric infarction. Decompressive surgery might be the most promising therapeutic option. For decisive answers, randomized, controlled clinical trials are needed. PMID- 12576975 TI - Interhospital variability in satisfaction with withdrawal of life support: room for improvement? AB - OBJECTIVE: To develop instruments to measure the current level of satisfaction with the withdrawal of life support among nurses and respiratory therapists and to use these instruments to determine whether differences in satisfaction levels with the withdrawal of life support could be found among different institutions. DESIGN: A multicenter, self-administered, questionnaire-based study. SETTING: The intensive care units of three academic and 11 community hospitals. SUBJECTS: A total of 412 critical care nurses and 117 respiratory therapists working in these intensive care units. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: After developing two similar questionnaires for critical care nurses and respiratory therapists, we assessed the reliability of the individual items for each questionnaire and only used those found to be reliable in the analyses. We found variability in the satisfaction level of critical care nurses among different intensive care units and among academic and community hospital intensive care units. Being involved in the plan for withdrawal of life support, comfort with the way discussions with the family had gone, comfort with patient sedation, and increasing experience with withdrawal of life support were items associated with increased nurse satisfaction. For respiratory therapists, there seemed to be similar differences among institutions in satisfaction level, although numbers were limited. Having the physician explain the plan to them and becoming more comfortable with withdrawal of life support as a result of increased experience were found to be associated with increased satisfaction levels for respiratory therapists. CONCLUSIONS: Satisfaction levels of intensive care unit nurses and respiratory therapists with the process of withdrawing life support vary among institutions. This variation may represent an opportunity for improvement in the process of withdrawing life support within this region. PMID- 12576976 TI - Circulating substances and energy metabolism in septic shock. PMID- 12576977 TI - Hormone replacement therapy for the critically ill. PMID- 12576978 TI - Granulocyte colony-stimulating factor for severe pneumonia: what do we do when the best laid plans for men (and mice and rats...) fail? PMID- 12576979 TI - NO way to warm the big toe. PMID- 12576980 TI - Shocking? PMID- 12576981 TI - Efficacy of lung recruiting maneuvers: it's all relative. PMID- 12576982 TI - Chaos, computers, and critical care. PMID- 12576983 TI - In critically ill patients with acute renal failure, outcomes, not dollars, should drive modality choice. PMID- 12576984 TI - Assessing futility of medical interventions--is it futile? PMID- 12576985 TI - How do we achieve adequate therapy for severe infection? PMID- 12576986 TI - Nicorandil: albuterol-like or cromolyn-like? PMID- 12576987 TI - Protein tyrosine kinase inhibitors as novel therapeutics for systemic inflammation, shock, and organ injury. PMID- 12576988 TI - Temperature regulation after cardiac arrest... Timing is everything! PMID- 12576989 TI - Can noninvasive ventilation succeed in the real world? The answer is YES! PMID- 12576990 TI - Does arginine become a "near" essential amino acid during sepsis? PMID- 12576991 TI - Evidence-based medicine: what do you do when there's no evidence? PMID- 12576992 TI - Procalcitonin measurement for differential diagnosis between pulmonary embolism and pneumonia. PMID- 12576993 TI - Hidden nutrition studies. PMID- 12576994 TI - Patient autonomy and the end of life. PMID- 12576995 TI - The last, best hope. PMID- 12576996 TI - Propylene glycol toxicity occurs during low-dose infusions of lorazepam. PMID- 12576997 TI - Ventilating asthmatics using intensive care ventilators. PMID- 12576998 TI - [Radiological technologists hope to increase activities in the industries]. PMID- 12576999 TI - [Image display devices (3)--LCD monitor]. PMID- 12577000 TI - [Emergent MR imaging of acute brain disease]. PMID- 12577001 TI - [Magnetic resonance. A preliminary knowledge]. PMID- 12577002 TI - [CT-pancreatography using various image processing technique]. PMID- 12577003 TI - [Image registration and fusion for multimodal medical imaging]. PMID- 12577005 TI - [A report of "Standard dosimetry of X-ray in diagnostic radiology"]. PMID- 12577004 TI - [A report of the Special Committee for the Future Planning on the JSRT: Future vision of the JSRT with the advanced education systems related to radiological technology]. PMID- 12577006 TI - [A report from a task group for construction of mammography database]. PMID- 12577007 TI - [Establishment and standardization of a quality assurance program for computed radiography systems]. PMID- 12577008 TI - [PET technology applied for FDG based examination--in consideration of the approval by the health insurance system--part 1]. PMID- 12577009 TI - [Patient skin dose in interventional radiology using radiochromic dosimetry film]. AB - Various types of X-ray examinations are currently being carried out for the purpose of diagnosis. However, since dose limits for contamination by medical examinations have not been set, management of dose measurements and contamination records is called for. With increasing use of the IVR technique, reports of radiation injury and the symptoms associated with it have become more common. To advance our understanding of this situation and to reduce contamination, it is necessary to carry out contamination management. The reflection film on which colors are formed by irradiating X-rays has recently come into use. Dose measurement is possible with the use of this film, and, because effective results can be obtained as a result of performing fundamental examinations, the film actually provides dose measurements for the IVR technique. Another benefit is that maximum patient skin dose and dose distribution can be determined in addition to dose measurement. Moreover, since various methods were examined in this study, the method of dose evaluation is also reported for those wishing to employ it in the clinical setting. PMID- 12577010 TI - [Significance of the visual evaluation method using the two-visit method (two sample preference test suggested by Ferris)]. AB - To increase the accuracy of the visual evaluation method, we studied the significance of the two-sample preference test using the two-visit method, as proposed by Ferris. Advantages of this method include not only its ability to estimate whether the image quality of the sample is statistically the same but also its ability to distinguish the difference between samples more clearly than when using the conventional two-sample preference test or paired-comparison method. However, a disadvantage is that this method involves two observations of the same sample. When using chest images, even if a difference between the samples is not recognizable by the conventional method, a significant difference can be elicited with this method. The data obtained by this method can be evaluated with the paired-comparison method, so some samples may be ranked as well. Therefore, we verified that the two-sample preference test, which Ferris formulated, is useful in visual evaluations. PMID- 12577011 TI - [Evaluation of gantry tilt correction in multi detector-row CT: effect on multi planar reconstruction images]. AB - In multi-slice CT, the scanning center of each line multiple-row detector array is shifted in the Y-axis direction by the tilting of the gantry. Correction of this effect is necessary when CT images are reconstructed. Correction of table feed is necessary as well. These corrections are possible with the Somatom Plus4 Volume Zoom. The use of multi-planar reconstruction (MPR) images has been increasing rapidly in the area of clinical diagnosis. Therefore, we examined the influence of MPR images according to gantry tilt. For each gantry tilt, the spatial resolution of MPR images was almost the same even though scanning pitch was changed. When the gantry was tilted, the full width at half maximum (FWHM) of the section sensitivity profile (SSP) was about 10% to 20% wider than that when the gantry was not tilted. The spatial resolution of MPR images is more affected by whether or not the gantry is tilted than by the angle of tilt. Gantry tilt correction is useful to obtain images without artifact for metal and artificial teeth; however, deterioration in spiral resolution cannot be avoided. PMID- 12577012 TI - [Three dimensional and four dimensional images for help to application]. PMID- 12577013 TI - [Three dimensional image for diagnosis and neurosurgical treatment--clinical application to navigational neurosurgery and neuroendovascular treatment]. PMID- 12577014 TI - [Statistical diagnostic radiology]. PMID- 12577015 TI - [Three-dimensional projection display of striatum dopamine function measured by PET]. AB - [(11)C]CFT and[(11)C]raclopride images obtained by positron emission tomography (PET) are used to evaluate pre-synaptic dopamine transporter availability and post-synaptic dopamine D(2) receptor binding, respectively. A combined study with these tracers is useful for the differential diagnosis of Parkinson's disease. We generated three-dimensional (3D) animations of striatum PET images for the diagnosis of Parkinson's disease. Brain images of a normal subject and a typical Parkinson's disease patient with[(11)C]CFT and[(11)C]raclopride were obtained using a PET camera. Three-dimensional animations were generated from serial maximum intensity projection (MIP) images created by gradually changing the projection angle. Furthermore, the striatum images extracted from brain data were superimposed over a brain surface magnetic resonance (MR) image that was created by the volume-rendering method, and 3D animations were similarly generated. The present 3D animations were clinically useful for the differential diagnosis of brain diseases, because we were able to observe distributions of[(11)C]CFT and[(11)C]raclopride from any angle and to grasp at a glance the regional differences of distributions in reference to anatomical landmarks. PMID- 12577016 TI - [Fundamental evaluation of segmented attenuation correction method for clinical FDG-PET studies: simulation of pulmonary mass lesions in phantom studies]. AB - OBJECTIVE: Both the segmented attenuation correction (SAC) method and post injection transmission scanning are useful and widespread in clinical whole-body FDG-PET studies. The SAC method usually accomplishes smoothing of the transmission data. This calculation segments a micro -map into three degrees (lung, soft tissue, and bone) of attenuation coefficient. This method is used to reduce transmission scan time without deteriorating the quality of PET images. However, the SAC method has a tendency to underestimate the attenuation coefficient, resulting lower detectability for lung field mass lesions. We therefore evaluated the quantitative accuracy of the SAC method using transmission scanning and emission scanning data in a phantom study. METHODS: A dedicated 3D PET scanner, the Siemens ECAT EXACT HR+, was used to scan images of two types of phantoms, a spherical phantom (Japan Radioisotope Association phantom) and a cylindrical phantom (20 cm in diameter). We evaluated differences between transmission images ( micro -map) of the SAC method and measured attenuation correction (MAC) method, these two kinds of attenuation-corrected emission data (emission + SAC method, emission + MAC method), and emission data only (without attenuation correction). RESULTS: In the micro -map, recovery coefficient (RC) values at 10 mm in diameter were 0.27 and 0.00 in the MAC and SAC methods, respectively, in the spherical hot area. For the emission data, the emission + SAC method and emission + MAC method showed almost the same RC values for all sizes of hot area diameter. The SAC method, however, resulted in 20% underestimation for all sizes of hot area diameter as compared with the MAC method. CONCLUSION: In pulmonary mass lesions, it is necessary to correct for the partial volume effect in quantitative PET measurement. However, from our data, the SAC method is not appropriate for partial volume effect correction. PMID- 12577017 TI - [Measurement of left ventricular chamber and myocardial volume in hypertrophic cardiomyopathy patients by ECG-gated myocardial perfusion SPECT: application of a newly developed edge-detection algorithm]. AB - Quantitative gated SPECT (QGS) software has been reported to demonstrate inaccurate edge detection in the left ventricular chamber in hypertrophic cardiomyopathy patients. In this study we developed a method to calculate left ventricular volume (LVV) and left myocardial volume (LMV) from gated SPECT data using a newly developed edge-detection algorithm, and we compared it with the QGS method of calculating LVV and LMV in a phantom study. Our method gave more accurate measurements LVV and LMV whereas the QGS method underestimated LMV. Compared with QGS LVV and LMV, our method yielded better results in the phantom study. PMID- 12577018 TI - [4D-MRI using the synchronized sampling method (SSM)]. AB - A synchronized sampling method (SSM) was developed for the study of voluntary movements by combining the electrocardiographic (ECG) gating method with an external triggering device, and four-dimensional magnetic resonance imaging (4D MRI) at a rate of 30 frames per second was accomplished by volumetric imaging with the SSM. This method was first applied to the motion imaging of articulatory organs during repetitions of a Japanese five-vowel sequence, and the dynamic change in vocal tract area function was demonstrated with sufficient temporal resolution. This paper describes the methodology, applicability, and limitations of 4D-MRI with the SSM. PMID- 12577019 TI - [A study of the influence scan timing and depiction of blood vessel diameter in CE-3DMRA: simulation by pulsatile flow]. AB - In imaging using contrast-enhanced three-dimensional magnetic resonance angiography (CE-3DMRA), optimizing the delay time from the start of intravenous injection of contrast medium to the start of scanning has generally been an important concern when obtaining blood vessel images of good contrast. Recent methods of mechanically assessing the attainment of contrast medium injection include Smart-Prep and others. Another method is the Test Bolus, in which a small amount of contrast medium determines the timing of scan start, in quest of the time intensity curve. Because these methods are not necessarily satisfactory, the corrected method is used in clinical cases. In terms of how scan timing affects blood vessel depiction, no study has examined pulsatile flow, which is carried out by simulation. On the other hand, there are reports on data filling of k space using imitation blood vessels on a computer and simple experimental equipment. This research examined experimentally blood vessel depiction according to scan timing and the diameter of blood vessels by using a systemic circulation simulator that incorporated hemodynamic circulation in which pulsatile flow is the same as that of a human body, using an artificial heart developed especially for MRI. It is thought that scan timing in CE-3DMRA affects the depiction of blood vessels for which the diameter of the blood vessel differs from the experimental result. Phase-encoding k-space data filling is the contrast-to-noise ratio (CNR) of each blood vessel irrespective of sequential and centric k-space ordering. The timing shift phenomenon, in which a blood vessel is so thin that the scan timing is overdue and shows a relatively high value, can occur. Moreover, that scan timing affects not only the diameter of a blood vessel but also depiction of the narrowing of a stenotic blood vessel or a branch blood vessel was demonstrated clinically and experimentally. Therefore, blood vessel depiction changes with scan timing, and it is clinically important to determine the time of arrival of contrast medium and the duration of the enhancement effect in quest of the time intensity curve by test bolus when clearly describing the target blood vessel. PMID- 12577020 TI - [Evaluation of three-dimensional contrast-enhanced MRA using differential rate k space sampling (DRKS)]. AB - The progress of three-dimensional (3D) magnetic resonance angiography used in combination with contrast medium (CE-MRA) has been remarkable. Currently, angiography aims at improvements in time resolution without sacrificing spatial resolution. We conducted a basic study of 3D differential rate k-space sampling (DRKS) in which the slice direction of the k-space is divided into two or more areas, the echo data near zero encoding is sampled by a higher time resolution than data in other areas, and reconstruction is done within a short time. This technique involved a problem in which ghost artifacts occur easily when the concentration of contrast medium changes extremely or when signal intensity changes suddenly. This is probably due to a difference in the time to sample data between the low- and high-frequency areas. When we used DRKS to grasp these characteristics, however, it was useful because it allowed reconstruction with an extremely high time resolution. PMID- 12577021 TI - [Evaluation of appearance with high-speed CE-3dMRDSA in brain]. AB - We evaluated the fitting scan technique for CE-3dMRDSA, and found common ground that determines spatial resolution and time resolution (slab thickness and partition number). We also examined the relation between appearance and time resolution (volume of contrast medium and injection speed). To obtain good image contrast, the volume of contrast medium needs to be at least 7 ml and suitable for an injection speed of 3-5 ml/sec. However, when we increased the volume of contrast medium and decreased injection speed, changes in MRDSA images with time became worse. The measure of the bolus with contrast medium was found to determine image contrast. When contrast medium is injected earlier, it circulates earlier within the brain. If the scan time is not short enough, it is not possible to observe changes in MRDSA images. And when spatial resolution is improved, time resolution becomes worse. Therefore, it is important to find the point of compromise between spatial resolution and time resolution. If we look for anterior MIP images, the CNR in the spatial resolution didn't change, when the slice thickness is more than 3 mm. Because, the partial volume effect decide the image contrast. However, unless the view is from the front, slice thickness influences spatial resolution. Therefore, when we view MIP images from the lateral direction, slice thickness must be set at less than 2 mm. Results indicated that, in CE-3dMRDSA with the fitting technique, slice thickness should be less than 3 mm, partition number 16-20, slab thickness 48 mm, contrast medium volume 7-10 ml, and injection speed 3-5 ml. PMID- 12577022 TI - [Evaluation of three-dimensional enhanced brain surface imaging using CT (3D surface CT angiography) and magnetic resonance imaging (3D surface MR angiography)]. AB - The optimal imaging conditions for 3D brain surface imaging by magnetic resonance imaging (MRI) and multi-slice CT were investigated. Visualization of the sulci, gyri, and veins on the brain's surface was also compared between 3D surface images acquired using multi-slice CT and conventional single-slice CT and MRI. Various imaging parameters, including slice thickness, dose, and matrix size, were evaluated using our original brain surface phantom and longitudinal direction evaluation phantom as well as images obtained from healthy volunteers. Subjects of the clinical study were patients with arteriovenous malformations and brain tumors who underwent CT-angiography at the same time as MR-angiography. The quality of 3D images of the brain surface is most strongly influenced by partial volume effects related to slice thickness. In multi-slice CT, a slice thickness of 0.5 mm can be employed to minimize the partial volume effect, providing results that are far superior to those that can be achieved by conventional single-slice 3D-CT. In addition, the excellent S/N of multi-slice CT permits the veins on the brain's surface to be clearly visualized without the use of contrast medium. With regard to visualization of the sulci and gyri, although some problems remain to be overcome, multi-slice CT was found to be equivalent to 3D surface imaging using MRI. PMID- 12577024 TI - [Volume rendering in head MRA: discrimination of aneurysms by low-threshold processing]. AB - When processing head MRA images, threshold values are automatically established using the MIP method, resulting in a high degree of reproducibility. As a result, two different individuals can produce highly comparable images. In addition, the MIP method is reportedly effective for depicting fine vessels. However, since information other than maximum values is ignored by the MIP method, data contained in original MRA images are not optimally utilized. The results confirmed that much of the information contained in original MRA images could not be seen on MIP processed images. In many cases, low-threshold processing was useful for depicting fine vessels and arterioles that could not be seen on MIP processed images. PMID- 12577023 TI - [Usefulness of three-dimensional MR images of brain tumors for surgical simulation]. AB - The purpose of this study was to determine the usefulness of three-dimensional (3D) MR imaging of brain tumors for surgical planning. Sixty-nine patients with various tumors of the brain were included in the present study. Using a volume rendering (VR) method on an independent workstation, 3D-MR images were obtained with the fast-SPGR sequence after Gd-DTPA administration. VR images could show an exact relationship between the surface of the brain and major vessels. However, in patients with deeply located tumors, VR images did not necessarily provide sufficient information as to the relationship between the tumor and vessels. In combination with a surface-rendering method, 3D-MR imaging could demonstrate the exact relationships among the tumors, major vessels, and surface of the brain. In tumors without contrast enhancement, this method was able to show 3D images of tumors with surrounding structures. For neurosurgeons, 3D-MR images were useful for understanding the surface anatomy and surrounding structures of the tumors prior to surgery. These images were also helpful in explaining the condition of the disease to patients and their families. PMID- 12577025 TI - [Actual CT inspection (2): the directions of contrast media, and window setting]. PMID- 12577026 TI - [Development of 4D-CT scanner using a 256-rows detector]. PMID- 12577027 TI - [Image processing workstation M900 QUADRA for multi slice CT]. PMID- 12577028 TI - [Technical application of three-dimensional visualization and measurement for breast cancer using multidetector-row CT scanner]. AB - This paper describes an integrated methodology that addresses the operation of 3D imaging and measurement for the diagnosis and surgical operation of breast cancer. The main system for breast CT imaging used a multi-slice CT scanner and 3D-workstation. The clinical sequence was performed in three phases with non contrast and biphasic contrast-enhanced studies that were performed with multi slice helical CT scanning. Acquired DICOM images were directly sent to the 3D workstation that implemented our developed functions. Four types of transfer functions (mammary skin, vessel, lymph node, and tumor) were set based on region of-interest (ROI) measurements. After that, two types of segmentation were prepared with volumetry and measurement of the size of breast cancer. Batch processing for 3D visualization and the measurement of breast tumors was helpful for diagnosis and surgical planning. PMID- 12577029 TI - [Transluminal imaging of three-dimensional CT and MR angiograms for transluminal observation of angioarchitecture of cerebral aneurysms]. AB - To investigate the angioarchitecture of cerebral aneurysms, we studied various opacity curves to select specific volume data of CT and MR angiograms from the opacity charts of CT density or MR signal intensity distribution. We developed the method of transluminal imaging of CT and MR angiograms; the vessels and aneurysms were depicted transluminally through spaces between the rings of the vessel walls. Two cases of unruptured cerebral aneurysms were studied by transluminal imaging of three-dimensional CT and MR angiograms. The technical aspects of transluminal imaging and characteristics of volume data, obtained by CT and MR angiograms, were discussed. PMID- 12577030 TI - [Usefulness of simulation with multi-slice CT for laparoscopic nephrectomy]. AB - In recent years, laparoscopic surgery has attracted attention as a minimally invasive type of surgery because of the small surgical wounds and early recovery it provides. We carry out this technique on the basis of volume data that we make use of in multi-slice CT imaging technology in laparoscopic nephrectomy by the retroperitoneal approach, and we have created CT virtual laparoscopy by virtual endoscopic display as an intra-operative navigatior with an image analysis system. We provide information on detailed vascular anatomy to form intra operative images that act as similar support images. With the provision of this volume data, we consider virtual endoscopic display the most suitable method for surgery. When we perform virtual laparoscopy, we simulate the insertion point and angle, the order of vascular structures and their locations, the number of arteries and veins, and their bifurcation points and ligation points in conjunction with the surgeon prior to operation. As the branch patterns of the renal artery are varied, perioperative confusion and surgical mishaps can be avoided through the information that is provided beforehand. Thus surgery is more accurate and proceeds more smoothly, because the surgeon has accurate anatomical information. In addition, the time required for surgery is decreased, reducing risk and the possibility of complications. PMID- 12577032 TI - [Concept of multi-functional LEONARDO workstation]. PMID- 12577031 TI - [Experimental study of pseudo-stenosis artifact on three-dimensional DSA]. AB - Rotational three-dimensional digital subtraction angiography (3D-DSA) is very useful for interventional neuroradiology, especially in the endovascular therapy of cerebral aneurysms. However, pseudo-stenosis artifact on the vessel, which runs vertically to the rotational axis, was observed clinically. In this study, this artifact was confirmed in an experiment with 4.5-millimeter diameter vessel phantoms. The attenuation of the phantom at each degree of exposure (44 directions) was measured on the workstation (DSA pixel value). DSA pixel values were plotted from data of 10, 20, 30, 40, and 50 millimeter lengths, respectively. The saturated DSA pixel value of tangent projection on phantoms of 30 millimeters or more in length was observed. This phenomenon induces pseudo stenosis artifact on 3D-DSA. The maximum reduction in the diameter of the phantom was 27.4% on the length of 50 millimeters. We confirmed that the two-dimensional data vertical to the rotational axis were inaccurate when a straight-coursed, long, segmented vessel was present. Under this special condition, vessels on 3D DSA were displayed as smaller than their actual diameter. PMID- 12577033 TI - [CT applications for GE advantage workstation]. PMID- 12577034 TI - [Data handling for multislice CT]. PMID- 12577035 TI - [Quick net]. PMID- 12577036 TI - [Routine workstation system]. PMID- 12577037 TI - [INTegrated ImAGEs for realistic 3D visualization]. PMID- 12577038 TI - [Technical approach to 3D visualization for breast cancer with plug'n view]. PMID- 12577039 TI - [Diffusion tensor imaging analysis using PC version Dr.View/LINUX]. PMID- 12577040 TI - [Functions of the latest 3D image processing system]. PMID- 12577041 TI - [Development of the workstation suitable for large scale, high speed, and high resolution image data processing, and a future view]. PMID- 12577042 TI - [Research and studies on design and production for medical diagnostic imaging systems considered environmental problems part 3]. PMID- 12577043 TI - Ethics and animal research. PMID- 12577045 TI - Lafutidine, a novel histamine H2-receptor antagonist, increases serum calcitonin gene-related peptide in rats after water immersion-restraint stress. AB - Lafutidine is a novel histamine H(2)-receptor antagonist with a potent and long lasting anti-acid secretory effect that has also been found to have a potent gastroprotective effect. We investigated the effect of lafutidine on gastric mucosal injury induced in rats with the use of water-immersion restraint stress (WRS) by examining serum calcitonin gene-related peptide (CGRP) concentrations, which we measured with the use of an enzyme immunometric assay. WRS-induced mucosal erosive injury in the stomach was reduced significantly by both lafutidine and famotidine pretreatment (from 7.79 +/- 2.02 mm(2) to 3.09 +/- 0.74 mm(2) and 4.05 +/- 1.18 mm(2), respectively). A single administration of lafutidine or famotidine did not change the serum CGRP concentration from the control value when these drugs were administered without WRS. Lafutidine pretreatment before WRS caused a significant increase in serum CGRP concentration compared with famotidine (lafutidine, 86.64 +/- 9.52 pg/mL; famotidine, 47.55 +/- 4.35 pg/mL; control, 58.43 +/- 6.07 pg/mL). Our results suggest that lafutidine augments CGRP release from the rat stomach when administered before the induction of WRS. PMID- 12577044 TI - Autosomal dominant polycystic kidney disease: molecular genetics and pathophysiology. AB - In autosomal dominant polycystic kidney disease (ADPKD), the precise steps leading to cyst formation and loss of renal function remain uncertain. Pathophysiologic studies have suggested that renal tubule epithelial cells form cysts as a consequence of increased proliferation, dedifferentiation, and transition to a secretory pattern of transepithelial-fluid transport. Since the cloning of two genes implicated in ADPKD, there has been an explosion of information about the functions of the gene products polycystin 1 and 2. In this review, we discuss what is known of the functions of the polycystins and how this information is providing important insights into the molecular pathogenesis of ADPKD. PMID- 12577046 TI - Neutron-activation analysis: a novel method for the assay of iohexol. AB - Accurate measurement of glomerular filtration rate (GFR) is of great importance in both research and clinical medicine. Available methods require radiation exposure or are technically difficult, thereby limiting their utility. Recent work has validated plasma clearance of nonradioactive contrast agents (iohexol and iothalamate) for the measurement of GFR. However, their clinical utility has been restricted by the difficulties associated with the detection of contrast agents in serum. In this investigation, we evaluate the sensitivity, accuracy and precision of neutron activation analysis (NAA) to measure serum iohexol at concentrations necessary for estimating GFR. We subjected aliquots of serum containing 0 to 6470 microg/mL iohexol to neutron activation by placing them in a neutron beam for 1 minute. The activation process resulted in the elevation of iohexol's naturally abundant iodine 127 to iodine 128. The spontaneous decay of (128)I to xenon 128 (proportional to the amount of total iodine in the sample) was calculated by means of spectrographic analysis. The correlation between the predetermined elemental mass of iodine in the sample and that measured on NAA was then determined. A similar analysis was performed to establish the intra- and interday accuracy and precision, with multiple measurements taken over a single day and over the course of a month. We noted excellent correlation between iodine measured on NAA and the known elemental mass (r(2) =.99). Measurements were highly accurate (mean accuracy 2.4% +/- 1.8%), with excellent intra- and interday reliability (mean coefficient of variation 4.1% +/- 1.6%). NAA is a feasible and reproducible method of detecting iohexol for the purpose of measuring GFR. PMID- 12577047 TI - Calcium-binding proteins MRP 8 and 14 in a Staphylococcus aureus infection model: role of therapy, inflammation, and infection persistence. AB - Concentrations of the calcium-binding proteins of the S100 family, myeloid related proteins 8 and MRP 14 (MRP8/14), are elevated in chronic infections, yet the role of these proteins is not clearly defined. Using commercial and developed enzyme immunoassays, we assayed for MRP8/14 in sterile-filtered abscess fluid from tissue-cage-implanted rats and rabbits. Staphylococcus aureus abscesses were created 6 weeks after the intraperitoneal implantation of tissue cages. Leukocytes, bacteria, and non-protein-bound calcium and zinc were measured in the infection exudate at day 3 or 5 of infection and after 8 days of treatment with antimicrobials beta-lactams (18 rabbits, 35 rats) and fluoroquinolone-rifampin (6 rabbits). Half of the infected rats were depleted of neutrophils; these rats exhibited significantly lower MRP 8/14 concentrations on all days sampled, regardless of the level of infection. The level of abscess MRP 8/14 is high early in the course of infection but decreases with effective antimicrobial treatment by as much as 100-fold. Thirty-day-old abscesses with log 6 bacterial counts and low neutrophil counts showed low concentrations of MRP 8/14 in these models. In abscess fluid, interleukin-6, as a representative marker of inflammation, correlated with MRP8/14, whereas ionized calcium and zinc did not. Our data suggest that infection and inflammation are not equal stimuli for MRP 8/14. The neutrophil appears to be the main source of MRP8/14 in this model. PMID- 12577048 TI - Deferoxamine promotes survival and prevents electrocardiographic abnormalities in the gerbil model of iron-overload cardiomyopathy. AB - We investigated the time course of electrocardiographic (ECG) changes in the Mongolian gerbil model of iron overload and the effects of the iron chelator deferoxamine (DFO) on these changes. Iron overload was produced with weekly subcutaneous injections of low doses (200 mg/kg/wk) or high doses (800 mg/kg/wk) of iron-dextran. DFO was administered subcutaneously at a dose of 200 mg/kg/day to high-dose animals. Our results show that (1) survival of iron-overloaded gerbils is dose-dependent, with median survival times of 68 and 14 weeks for low- and high-dose animals, respectively; (2) both low and high doses produce prolongation of the PR interval and bradycardia in early stages and prolongation of the QT interval, premature ventricular contractions, variable degrees of atrioventricular block, changes in the ST segment, and T-wave inversion at later stages coinciding with the development of heart failure; (3) DFO prevented death during 20 weeks of high-dose iron-dextran; (4) DFO prevented ECG changes, although delayed prolongation of PR intervals and QRS complexes occurred; and (5) despite marked prolongation of survival and prevention of ECG changes, DFO had modest effects on total cardiac iron content. We speculate that DFO chelates a small iron pool located within the cytoplasm of iron-overloaded cardiomyocytes. PMID- 12577050 TI - Gene transfer in purified human hematopoietic peripheral-blood stem cells by means of electroporation without prestimulation. AB - Gene transfer in hematopoietic stem cells (HSCs) is an important tool, exploring regulation of the hematopoietic system and understanding the development and expansion of malignant cell clones. It is also a mandatory step for gene therapy of hematopoietic disorders. Although retroviral transduction of HSCs is effective, prestimulation of cells is generally required, also inducing differentiation of HSCs. Furthermore, the risk of viral recombination and insertional mutagenesis cannot be ruled out. Potential advantages of nonviral transfection are biosafety and easy management. However, experience in nonviral methods for transfecting peripheral-blood stem cells (PBSCs) is limited. To avoid differentiation, we evaluated the efficiency of gene transfer by means of electroporation without cytokine prestimulation. Compared with prestimulated (stem-cell factor, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, interleukin-3, interleukin-6, erythropoietin, and monoclonal antibody to transforming growth factor-beta), transfection of thawed nonstimulated PBSCs was equally efficient, with a median transfection rate of 3.7%, transfection efficiency of 0.8%, and survival of 19.5% (n = 5). With freshly isolated HSCs, the rate of transfected cells could be increased to a median of 27.0% (range 8.3%-31.0%), transfection efficiency of 6.9% (range 4.5% 12.6%), and survival of 43% (range 22%-64%) (n = 5). However, the percentage of transfected cells declined with time; almost no cells were detectable by day 11. One cause for the lack of long-term expression of the heterologous gene in this system was induction of apoptosis in transgenic PBSCs, shown by up-regulation of CD95 (FAS antigen). PMID- 12577049 TI - Serum noncholesterol sterols during inhibition of cholesterol synthesis by statins. AB - We studied changes in serum cholestanol and plant sterols (indexes of cholesterol absorption) and cholesterol precursors (indexes of cholesterol synthesis) in response to cholesterol reduction by way of 1 year's treatment with atorvastatin (n = 102) and simvastatin (n = 105) treatments in patients with coronary heart disease. Serum cholesterol levels and ratios of the precursor sterols to cholesterol after 1 year of treatment were reduced in proportion to the pretreatment values (33% +/- 1% by simvastatin and 36% +/- 1% by atorvastatin; P <.01 for difference between groups) for cholesterol; the respective reductions in the precursor sterol:cholesterol ratios were also higher with atorvastatin (50% +/- 2% for lathosterol) than with simvastatin (42% +/- 1%; P <.01 between groups), but the ratio of squalene to cholesterol was increased (17% +/- 5%, P <.001) by atorvastatin. Plant sterol concentrations were gradually increased by atorvastatin but decreased initially by simvastatin. However, their ratios with respect to cholesterol were increased by as much as 82% with atorvastatin and by as much as 39% with simvastatin. In conclusion, effective inhibition of cholesterol synthesis and subsequent reduction in serum cholesterol levels by statins lead to increases in serum plant-sterol levels, probably as a result of reduced biliary secretion and enhanced absorption of these sterols. Because serum plant sterols have been claimed to be involved in the early development of atherosclerosis, the question arises whether continuously increasing serum plant sterols during long-term statin treatment should be prevented by cholesterol malabsorption (eg, by plant stanol ester consumption), especially in subjects with high baseline plant sterol values and effective sterol absorption. PMID- 12577051 TI - Structure of a gametocyte protein essential for sexual development in Plasmodium falciparum. AB - Malaria transmission is dependent on the development of sexual forms of Plasmodium falciparum, called gametocytes, in the vertebrate host. Pfg27 is an abundantly expressed sexual stage-specific protein that is essential for gametocytogenesis in P. falciparum. We describe the crystal structure of Pfg27, which reveals a novel fold composed of two pseudo dyad-related repeats of the helix-turn-helix motif. Structurally equivalent helices of each repeat either form a dimer interface or interact with RNA in vitro. One side of the dimer presents an unprecedented juxtaposition of four polyproline (PXXP) motifs. Preliminary binding data indicate that these sites are capable of binding Src homology-3 (SH3) modules. Molecular modeling suggests that the dimer can accommodate two SH3 modules simultaneously, potentially enabling molecular crosstalk between SH3-containing proteins. The structural and initial biochemical evidence suggests that Pfg27 may serve as a platform for RNA and SH3 binding. PMID- 12577052 TI - Crystal structure of a MARCKS peptide containing the calmodulin-binding domain in complex with Ca2+-calmodulin. AB - The calmodulin-binding domain of myristoylated alanine-rich C kinase substrate (MARCKS), which interacts with various targets including calmodulin, actin and membrane lipids, has been suggested to function as a crosstalk point among several signal transduction pathways. We present here the crystal structure at 2 A resolution of a peptide consisting of the MARCKS calmodulin (CaM)-binding domain in complex with Ca2+-CaM. The domain assumes a flexible conformation, and the hydrophobic pocket of the calmodulin N-lobe, which is a common CaM-binding site observed in previously resolved Ca2+-CaM-target peptide complexes, is not involved in the interaction. The present structure presents a novel target recognition mode of calmodulin and provides insight into the structural basis of the flexible interaction module of MARCKS. PMID- 12577053 TI - Rad54 protein possesses chromatin-remodeling activity stimulated by the Rad51 ssDNA nucleoprotein filament. AB - In Saccharomyces cerevisiae, the Rad54 protein participates in the recombinational repair of double-strand DNA breaks together with the Rad51, Rad52, Rad55 and Rad57 proteins. In vitro, Rad54 interacts with Rad51 and stimulates DNA strand exchange promoted by Rad51 protein. Rad54 is a SWI2/SNF2 related protein that possesses double-stranded DNA-dependent ATPase activity and changes DNA topology in an ATP hydrolysis-dependent manner. Here we show that Rad54 catalyzes bidirectional nucleosome redistribution by sliding nucleosomes along DNA. Nucleosome redistribution is greatly stimulated by the Rad51 nucleoprotein filament but does not require the presence of homologous single stranded DNA within the filament. On the basis of these data, we propose that Rad54 facilitates chromatin remodeling and, perhaps more generally, protein clearing at the homology search step of genetic recombination. PMID- 12577054 TI - Defective thymocyte apoptosis and accelerated autoimmune diseases in TRAIL-/- mice. AB - TRAIL, the tumor necrosis factor-related apoptosis-inducing ligand, selectively induces apoptosis of tumor cells, but not most normal cells. Its role in normal, nontransformed tissues is not clear. We report here that mice deficient in TRAIL have a severe defect in thymocyte apoptosis-thus, thymic deletion induced by T cell receptor ligation is severely impaired. TRAIL-deficient mice are also hypersensitive to collagen-induced arthritis and streptozotocin-induced diabetes and develop heightened autoimmune responses. Thus, TRAIL mediates thymocyte apoptosis and is important in the induction of autoimmune diseases. PMID- 12577055 TI - TCR ligand discrimination is enforced by competing ERK positive and SHP-1 negative feedback pathways. AB - Functional discrimination between structurally similar self and foreign antigens is a main attribute of adaptive immunity. Here we describe two feedback mechanisms in T lymphocytes that together sharpen and amplify initial signaling differences related to the quality of T cell receptor (TCR) engagement. Weakly binding ligands predominantly trigger a negative feedback loop leading to rapid recruitment of the tyrosine phosphatase SHP-1, followed by receptor desensitization through inactivation of Lck kinase. In contrast, strongly binding ligands efficiently activate a positive feedback circuit involving Lck modification by ERK, preventing SHP-1 recruitment and allowing the long-lasting signaling necessary for gene activation. The characteristics of these pathways suggest that they constitute an important part of the mechanism allowing T cells to discriminate between self and foreign ligands. PMID- 12577056 TI - Homozygous loss of ICOS is associated with adult-onset common variable immunodeficiency. AB - No genetic defect is known to cause common variable immunodeficiency (CVID), a heterogeneous human disorder leading to adult-onset panhypogammaglobulinemia. In a search for CVID candidate proteins, we found four of 32 patients to lack ICOS, the "inducible costimulator" on activated T cells, due to an inherited homozygous deletion in the ICOS gene. T cells from these individuals were normal with regard to subset distribution, activation, cytokine production and proliferation. In contrast, naive, switched and memory B cells were reduced. The phenotype of human ICOS deficiency, which differs in key aspects from that of the ICOS-/- mouse, suggests a critical involvement of ICOS in T cell help for late B cell differentiation, class-switching and memory B cell generation. PMID- 12577057 TI - Chromosome 6q25 is linked to susceptibility to leprosy in a Vietnamese population. AB - Leprosy, a chronic infectious disease caused by Mycobacterium leprae, affects an estimated 700,000 persons each year. Clinically, leprosy can be categorized as paucibacillary or multibacillary disease. These clinical forms develop in persons that are intrinsically susceptible to leprosy per se, that is, leprosy independent of its specific clinical manifestation. We report here on a genome wide search for loci controlling susceptibility to leprosy per se in a panel of 86 families including 205 siblings affected with leprosy from Southern Vietnam. Using model-free linkage analysis, we found significant evidence for a susceptibility gene on chromosome region 6q25 (maximum likelihood binomial (MLB) lod score 4.31; P = 5 x 10(-6)). We confirmed this by family-based association analysis in an independent panel of 208 Vietnamese leprosy simplex families. Of seven microsatellite markers underlying the linkage peak, alleles of two markers (D6S1035 and D6S305) showed strong evidence for association with leprosy (P = 6.7 x 10(-4) and P = 5.9 x 10(-5), respectively). PMID- 12577058 TI - Coordination of the random asynchronous replication of autosomal loci. AB - Random monoallelic expression and asynchronous replication define an unusual class of autosomal mammalian genes. We show that every cell has randomly chosen either the maternal or paternal copy of each given autosome pair, such that alleles of these genes scattered across the chosen chromosome replicate earlier than the alleles on the homologous chromosome. Thus, chromosome-pair non equivalence, rather than being limited to X-chromosome inactivation, is a fundamental property of mouse chromosomes. PMID- 12577059 TI - Germline mutations in PRKCSH are associated with autosomal dominant polycystic liver disease. AB - Polycystic liver disease (PCLD, OMIM 174050) is a dominantly inherited condition characterized by the presence of multiple liver cysts of biliary epithelial origin. Fine mapping established linkage to marker D19S581 (Z(max) = 9.65; theta = 0.01) in four large Dutch families with PCLD. We identified a splice-acceptor site mutation (1138-2A-->G) in PRKCSH in three families, and a splice-donor site mutation (292+1G-->C) in PRKCSH segregated completely with PCLD in another family. The protein encoded by PRKCSH, here named hepatocystin, is predicted to localize to the endoplasmic reticulum. These findings establish germline mutations in PRKCSH as the probable cause of PCLD. PMID- 12577060 TI - Evidence for co-evolution of gene order and recombination rate. AB - There is increasing evidence in eukaryotic genomes that gene order is not random, even allowing for tandem duplication. Notably, in numerous genomes, genes of similar expression tend to be clustered. Are there other reasons for clustering of functionally similar genes? If genes are linked to enable genetic, rather than physical clustering, then we also expect that clusters of certain genes might be associated with blocks of reduced recombination rates. Here we show that, in yeast, essential genes are highly clustered and this clustering is independent of clustering of co-expressed genes and of tandem duplications. Adjacent pairs of essential genes are preferentially conserved through evolution. Notably, we also find that clusters of essential genes are in regions of low recombination and that larger clusters have lower recombination rates. These results suggest that selection acts to modify both the fine-scale intragenomic variation in the recombination rate and the distribution of genes and provide evidence for co evolution of gene order and recombination rate. PMID- 12577061 TI - Long-term depression induced by sensory deprivation during cortical map plasticity in vivo. AB - Cortical map plasticity is thought to involve long-term depression (LTD) of cortical synapses, but direct evidence for LTD during plasticity or learning in vivo is lacking. One putative role for LTD is in the reduction of cortical responsiveness to behaviorally irrelevant or unused sensory stimuli, a common feature of map plasticity. Here we show that whisker deprivation, a manipulation that drives map plasticity in rat somatosensory cortex (S1), induces detectable LTD-like depression at intracortical excitatory synapses between cortical layer 4 (L4) and L2/3 pyramidal neurons. This synaptic depression occluded further LTD, enhanced LTP, was column specific, and was driven in part by competition between active and inactive whiskers. The synaptic locus of LTD and these properties suggest that LTD underlies the reduction of cortical responses to deprived whiskers, a major component of S1 map plasticity. PMID- 12577062 TI - Activity level controls postsynaptic composition and signaling via the ubiquitin proteasome system. AB - Experience-dependent remodeling of the postsynaptic density (PSD) is critical for synapse formation and plasticity in the mammalian brain. Here, in cultured rat hippocampal neurons, I found long-lasting, global changes in the molecular composition of the PSD dictated by synaptic activity. These changes were bidirectional, reversible, modular, and involved multiple classes of PSD proteins. Moreover, activity-dependent remodeling was accompanied by altered protein turnover, occurred with corresponding increases or decreases in ubiquitin conjugation of synaptic proteins and required proteasome-mediated degradation. These modifications, in turn, reciprocally altered synaptic signaling to the downstream effectors CREB (cyclic AMP response element binding protein) and ERK MAPK (extracellular signal regulated kinase-MAP kinase). These results indicate that activity regulates postsynaptic composition and signaling through the ubiquitin-proteasome system, providing a mechanistic link between synaptic activity, protein turnover and the functional reorganization of synapses. PMID- 12577063 TI - Elimination and strengthening of glycinergic/GABAergic connections during tonotopic map formation. AB - Synapse elimination and strengthening are central mechanisms for the developmental organization of excitatory neuronal networks. Little is known, however, about whether these processes are also involved in establishing precise inhibitory circuits. We examined the development of functional connectivity before hearing onset in rats in the tonotopically organized, glycinergic pathway from the medial nucleus of the trapezoid body (MNTB) to the lateral superior olive (LSO), which is part of the mammalian sound localization system. We found that LSO neurons became functionally disconnected from approximately 75% of their initial inputs, resulting in a two-fold sharpening of functional topography. This was accompanied by a 12-fold increase in the synaptic conductance generated by maintained individual inputs. Functional elimination of MNTB-LSO synapses was restricted to the period when these glycinergic/GABAergic synapses are excitatory. These results provide new insights into the mechanisms by which precisely organized inhibitory circuits are established during development. PMID- 12577064 TI - RhoD regulates endosome dynamics through Diaphanous-related Formin and Src tyrosine kinase. AB - Early endosomes move bidirectionally between the cell periphery and the interior through a mechanism regulated by the low molecular weight GTPase RhoD. Here, we identify a novel splice variant of human Diaphanous, hDia2C, which specifically binds to RhoD and is recruited onto early endosomes. Expression of RhoD and hDia2C induces a striking alignment of early endosomes along actin filaments and reduces their motility. This activity depends on the membrane recruitment and activation of c-Src kinase, thus uncovering a new role in endosome function. Our results define a novel signal transduction pathway, in which hDia2C and c-Src are sequentially activated by RhoD to regulate the motility of early endosomes through interactions with the actin cytoskeleton. PMID- 12577065 TI - A Ran signalling pathway mediated by the mitotic kinase Aurora A in spindle assembly. AB - The activated form of Ran (Ran-GTP) stimulates spindle assembly in Xenopus laevis egg extracts, presumably by releasing spindle assembly factors, such as TPX2 (target protein for Xenopus kinesin-like protein 2) and NuMA (nuclear-mitotic apparatus protein) from the inhibitory binding of importin-alpha and -beta. We report here that Ran-GTP stimulates the interaction between TPX2 and the Xenopus Aurora A kinase, Eg2. This interaction causes TPX2 to stimulate both the phosphorylation and the kinase activity of Eg2 in a microtubule-dependent manner. We show that TPX2 and microtubules promote phosphorylation of Eg2 by preventing phosphatase I (PPI)-induced dephosphorylation. Activation of Eg2 by TPX2 and microtubules is inhibited by importin-alpha and -beta, although this inhibition is overcome by Ran-GTP both in the egg extracts and in vitro with purified proteins. As the phosphorylation of Eg2 stimulated by the Ran-GTP-TPX2 pathway is essential for spindle assembly, we hypothesize that the Ran-GTP gradient established by the condensed chromosomes is translated into the Aurora A kinase gradient on the microtubules to regulate spindle assembly and dynamics. PMID- 12577066 TI - Homologous recombination in human embryonic stem cells. AB - Homologous recombination applied to mouse embryonic stem (ES) cells has revolutionized the study of gene function in mammals. Although most often used to generate knockout mice, homologous recombination has also been applied in mouse ES cells allowed to differentiate in vitro. Homologous recombination is an essential technique if human ES cells are to fulfill their promise as a basic research tool. It also has important implications for ES cell-based transplantation and gene therapies. Significant differences between mouse and human ES cells have hampered the development of homologous recombination in human ES cells. High, stable transfection efficiencies in human ES cells have been difficult to achieve, and, in particular, electroporation protocols established for mouse ES cells work poorly in human ES cells. Also, in contrast to their murine counterparts, human ES cells cannot be cloned efficiently from single cells, making it difficult to screen for rare recombination events. Here we report an electroporation approach, based on the physical characteristics of human ES cells, that we used to successfully target HPRT1, the gene encoding hypoxanthine phosphoribosyltransferase-1 (HPRT1), and POU5F1, the gene encoding octamer-binding transcription factor 4 (Oct4; also known as POU domain, class 5, transcription factor 1 (POU5F1)). PMID- 12577067 TI - A proteomics strategy to elucidate functional protein-protein interactions applied to EGF signaling. AB - Mass spectrometry-based proteomics can reveal protein-protein interactions on a large scale, but it has been difficult to separate background binding from functionally important interactions and still preserve weak binders. To investigate the epidermal growth factor receptor (EGFR) pathway, we employ stable isotopic amino acids in cell culture (SILAC) to differentially label proteins in EGF-stimulated versus unstimulated cells. Combined cell lysates were affinity purified over the SH2 domain of the adapter protein Grb2 (GST-SH2 fusion protein) that specifically binds phosphorylated EGFR and Src homologous and collagen (Shc) protein. We identified 228 proteins, of which 28 were selectively enriched upon stimulation. EGFR and Shc, which interact directly with the bait, had large differential ratios. Many signaling molecules specifically formed complexes with the activated EGFR-Shc, as did plectin, epiplakin, cytokeratin networks, histone H3, the glycosylphosphatidylinositol (GPI)-anchored molecule CD59, and two novel proteins. SILAC combined with modification-based affinity purification is a useful approach to detect specific and functional protein-protein interactions. PMID- 12577068 TI - A genetic approach to identifying mitochondrial proteins. AB - The control of intricate networks within eukaryotic cells relies on differential compartmentalization of proteins. We have developed a method that allows rapid identification of novel proteins compartmentalized in mitochondria by screening large-scale cDNA libraries. The principle is based on reconstitution of split enhanced green fluorescent protein (EGFP) by protein splicing of DnaE derived from Synechocystis sp. PCC6803. The cDNA libraries are expressed in mammalian cells following infection with retrovirus. If a test protein contains a functional mitochondrial targeting signal (MTS), it translocates into the mitochondrial matrix, where EGFP is then formed by protein splicing. The cells harboring this reconstituted EGFP are screened rapidly by fluorescence-activated cell sorting, and the cDNAs are isolated and identified from the cells. The analysis of 258 cDNAs revealed various MTSs, among which we identified new transcripts corresponding to mitochondrial proteins. This method should provide a means to map proteins distributed within intracellular organelles in a broad range of different tissues and disease states. PMID- 12577070 TI - Safety and cross infection. PMID- 12577072 TI - Further statistics in dentistry Part 8: Systematic reviews and meta-analyses. AB - A systematic review of research evidence is an efficient approach to integrating existing information, invariably a multiplicity of published articles, with a view to establishing whether the scientific findings are consistent. If so, it may be possible to draw conclusions and make recommendations about treatment regimens or observed effects which have greater credence than those obtained from individual studies. The systematic review relies on a specified checklist which determines which articles should be included in the review, and how each should be critically appraised to provide relevant information relating to the focus of the review. PMID- 12577073 TI - Development of a treatment outcome standard as a result of a clinical audit of the outcome of fixed appliance therapy undertaken by hospital-based consultant orthodontists in the UK. AB - Bristol's much-publicised cardiac surgery problems and subsequent enquiry(1) have drawn attention to the need for audit of treatment outcomes throughout all hospital specialties. Patient anxiety, government policy and the desire of the professions to re-establish public confidence, have further encouraged changes to the system. For medical and dental specialties, such challenges have already been taken up by the Royal Colleges with the establishment of clinical effectiveness committees. Hospitals have modified their procedures and, for consultants, yearly appraisal is already a reality. The Orthodontic Clinical Effectiveness Working Party of the Royal College of Surgeons of England (now the Clinical Effectiveness Committee of the British Orthodontic Society) set up this audit to measure the outcome of fixed appliance treatment and to establish a benchmark for the standard of treatment to be expected from a consultant orthodontist. This paper describes how the audit was carried out, presents the findings and goes on to discuss some of the wider issues involved in audit, clinical governance and appraisal. The Consultant Orthodontists Group of the British Orthodontic Society funded this audit and the results and data set of dental casts remain their property. PMID- 12577077 TI - Oral diseases and socio-economic status (SES). AB - OBJECTIVE: To determine the association between social, economic and behavioural risk factors and national prevalences of: oral cancer, dental caries (12-year olds) and destructive periodontal disease (35-44-year-olds). DATA SOURCES: Sources for the social and economic parameters were the UN Development Program; the behavioural risk factors' source was the World Health Organization, the UN Food and Agricultural Organization and the World Atlas of History. Oral diseases data came from UICC Globocan and the World Health Organization databases. DATA EXTRACTION: Data were extracted by hand from official publications. DATA SYNTHESIS: Data were synthesized and analyzed in sequence using SPSS, Pearson's correlation coefficient and multiple regression analyses. CONCLUSIONS: There is a discernable association between the three oral diseases and the variables selected, which varies in strength, being strongest for chronic destructive periodontitis and weakest for oral cancer. Dental caries lies in between. The degree to which variables account for differences in the three oral diseases between the countries studied is striking, being insignificant for oral cancer incidence, modest for oral cancer mortality, stronger for dental caries and strongest for destructive periodontal disease. Removing variables with strong co linearity with the Human Development Index has little effect on the regression coefficients. PMID- 12577078 TI - Sequential 3D X-ray microtomographic measurement of enamel and dentine ablation by an Er:YAG laser. AB - OBJECTIVE: To demonstrate the progression of crater growth during repeated sequential application of an Er:YAG laser to enamel and dentine, monitored using X-ray microtomography (XMT). DESIGN: A single centre study in which laser craters were created in blocks cut from human enamel and/or dentine under standardised and known conditions and then studied using XMT to obtain visualisation and quantification of the effects. SETTING: University setting, UK, 2001. MAIN OUTCOME MEASURES: Success was judged by an ability to obtain useful 3D XMT reconstructions of the blocks during crater development, and to make measurements from these data. These measurements were compared with data obtained from similar studies using different measurement techniques. RESULTS: Time sequences of 2D and 3D images were obtained which demonstrated the progression of laser craters in enamel and dentine. Quantitative measurements from these data enabled values to be derived for the rate of progression of crater depth per unit energy, and the volume of hard tissue removed per unit energy. These values were compared with data derived from other studies and shown to be broadly comparable. However, the present study is unique in that these values were obtained from a series of measurements of the same craters over time. CONCLUSIONS: 3D X-ray microtomography is shown to be a useful tool for quantitative measurements in dental research. For the Er:YAG laser, the relationship of laser crater depth and volume of mineral removed to applied energy was found to be linear. PMID- 12577084 TI - Cerebrovascular disease in type 2 diabetes mellitus. AB - Incidence of CVD in diabetic men was reported to be twice as that of non diabetics and almost three times greater in diabetic women in the Framingham Study. It is postulated that excessive glycation and oxidation, endothelial dysfunction and increased platelet aggregation may be responsible for endothelial proliferation and thickening of plasmatic membrane in small blood vessels ('lipohyalinosis') leading to lacunar infarction. Prothrombotic state may precipitate a stroke, however, platelet aggregability, elevated fibrinopeptide A (FPA) and D-dimer were not significantly related to stroke in diabetic mellitus (DM), whereas suppressed fibrinolytic activity was a common finding. Of many unknown factors in pathogenesis, the deficient insulin secretion, resistance to action of insulin at level of 'insulin receptors', changes in counter regulatory hormones (e.g. glucagon, pancreatic polypetides, growth hormone, catecholamines, etc.) and decrease in the hepatic sensitivity to insulin action in suppressing glucose output have received more attention. Hyperosmolar state can simulate stroke syndromes. Early recognition and treatment of risk factors such as hypertension or better glycemic control, correction of hyperlipidemia or obesity in diabetic population are important. In diabetic subjects already showing recurrent transient cerebral ischemic attacks (TIAs) or minor strokes, the benefit of antiplatelet agents or antithrombotic therapy in prevention of major strokes is well established. Ramipril has been found to be effective in reducing stroke risk by 33% in diabetic patinets in HOPE study. PMID- 12577085 TI - Management of congenital atlanto-axial dislocation: some lessons learnt. AB - Congenital atlantoaxial dislocation (AAD) has a high incidence in India. In these patients, even a minor trauma may precipitate severe morbidity. The management of mobile AAD consists of posterior stabilization. In fixed AAD, the offending compressive element is present anterior to the cervicomedullary junction, which should be generously removed by the transoral approach. In many of these patients, the assimilated posterior arch of atlas or the incurving posterior margin of the foramen magnum or associated Chiari I malformation also adds to the compromise of the canal diameter at the foramen magnum. Thus, in cases of fixed AAD, the hour glass appearance of the spinal canal at the craniovertebral junction should be converted into a funnel shaped appearance which is close to normal. Since these procedures require a generous removal of the osteoligamentous structures, posterior fusion should be done in all cases after decompressive surgery. In this review, the observations that emerged during the management of congenital AAD are presented with a special focus on the assessment scales modified to the Indian settings, hypermobile AAD, rotary C1-2 dislocation, and AAD associated with Chiari I malformation and syringomyelia. PMID- 12577086 TI - Tetanus. AB - Tetanus is a potentially life threatening disease affecting nearly 50,000 to 1 million people world wide every year. Four major clinical forms of tetanus are described i.e. generalized, cephalic, localized and neonatal. Neonatal tetanus is particularly common in developing countries, due to unhygienic child birth practices, social taboos and improper immunization of pregnant mothers. Management of this disorder involves a team approach and aims at eradicating focus of infection, neutralizing the toxin, controlling spasms and dysautonomia and providing adequate ventilatory and supportive care. Metronidazole may be the preferred antibiotic although penicillin is still used frequently. Adequate wound debridement is necessary to prevent spore germination. Spasms are usually managed by sedatives like diazepam and neuromuscular blocking agents. Magnesium sulphate is an attractive substitute and may be tried if ventilatory facilities are unavailable. Use of baclofen is potentially advantageous but cannot be routinely prescribed. Dysautonomia is difficult to manage and requires therapy with benzodiazepines, morphine, magnesium sulphate, adrenergic blockers and recently tried baclofen therapy. Supportive care including ventilatory assistance are highly essential for successful outcome of the patients. It is imperative that complications are diagnosed early and managed appropriately. Immunization is extremely effective and is the key to prevention. Adequate steps and measures should be taken to increase awareness of this potentially preventable disease. PMID- 12577087 TI - Multifocal motor neuropathy. PMID- 12577088 TI - Landau Kleffner syndrome: electroclinical and etiopathogenic heterogeneity. AB - Landau - Kleffner syndrome is a rare, functional, age-related epilepsy with aphasia and epileptiform discharges on EEG. The heterogenity of clinical presentations, course, long-term outcome and response to treatment suggests multiple underlying etiologies. Normal children abruptly develop deterioration of language functions along with spike and wave discharges on EEG. Clinical seizures may or may not be present. The aphasia responds poorly to most drugs. Valproic acid and benzodiazepines are most effective. Steroids and intravenous immunoglobulins have shown a variable response. Long-term outcome of aphasia is variable, many patients persist with residual impairment. Important questions regarding etiopathogenesis are unanswered. PMID- 12577089 TI - Primary central nervous system lymphoma: experience of 46 cases with review of literature. AB - Primary central nervous system lymphomas (PCNSL) constitutes only 1.0 to 1.5% of all brain tumors. Their incidence has gone up over tenfold in the last 25 years. Though, there has been an association of PCNSL with acquired immune deficiency syndrome (AIDS), yet the increased incidence of PCNSL appears to be real and unrelated to AIDS and organ transplantation. This increased incidence could be because of improvement in diagnostic technology and practice. The outcome remains gloomy despite surgical resection, radiotherapy and intensive adjuvant chemotherapy regimens, as majority of the patients succumb to the disease, with only 30-40% survival in patients under 70 years of age. PMID- 12577090 TI - Long term outcome in patients with severe head injury and bilateral fixed dilated pupils. AB - Patients with severe head injury with bilateral dilated unreactive pupils are considered to have a grave prognosis. Hence proper planning and aggressive management becomes mandatory for achieving good results. We present the outcome of consecutive 166 patients with severe head injury, admitted between January 1996 and December 2000 and analysed retrospectively. All the patients had an initial GCS of 8 or less and post resuscitation bilateral dilated unreactive pupils. Our aim was to analyze the long term outcome in these patients and identify the other significant prognostic factors. Of the 166 patients, 42 (25.30%) had a functional outcome (good recovery in 10.24%, moderate disability in 15.06%), and 124 (74.69%) had a poor outcome (death in 58.43% and severe disability in 16.26% of cases). There were 45 patients with polytrauma and 24 of these patients (53.33%) succumbed to the injuries. Obliteration of the basal cisterns and contusion were the common CT scan findings. Factors adversely affecting the survival included age of the patient, polytrauma with shock, initial GCS of 3 or 4, and compression of the basal cisterns on the initial CT scans. At follow up, most of the patients with a functional outcome showed a significant improvement in their motor function but continued to have neuro behavioral and cognitive deficits. PMID- 12577091 TI - In vivo temperature measurements in brain tumors using proton MR spectroscopy. AB - The chemical shift difference between the water resonance and the methyl resonance of N-acetylaspartate was used to determine the in vivo temperature in 43 patients with histologically proven brain tumors. Temperatures were also estimated from the contralateral side in 15 of these patients. There was a significant difference (p < 0.05) ] in temperature between meningiomas and the other tumors viz. low grade astrocytomas, grade IV astrocytomas and oligoastrocytomas. Temperature was also significantly different between the contralateral side and the meningiomas (p < 0.05). PMID- 12577092 TI - Correlation of ictal EEG and SPECT studies in patients of intractable epilepsy with normal MRI. AB - Identification of the epileptic focus is the most important requirement for a successful surgical outcome in intractable epilepsy. Patients with normal MRI on high-resolution imaging pose a significant challenge in this regard. We tried to identify the epileptic focus using interictal / ictal SPECT and ictal EEG patterns in 14 patients with normal MRI. Rhythmic ictal onset activity helped to identify a single focus in 10 patients (71.43%). Inter ictal SPECT detected a focus corresponding to ictal EEG in 6 out of 10 patients (60%) in whom the test was performed. Ictal SPECT done in 5 patients correctly identified the focus in all patients (100%). Our data indicate that it is possible to obtain localization in patients with normal MRI using a combination of ictal EEG patterns and an ictal SPECT study. PMID- 12577093 TI - Glioneuronal migration and development disorders: histological and immunohistochemical study with a comment on evolution. AB - Glioneuronal migration disorders of the brain evolve primarily due to aberration in neuronal migration, maturation and programming in the development of various topographic zones in the brain, following pathological alterations in glial and neuronal interactions. These are broadly referred as cortical dysplastic conditions. While these dysplastic conditions involving cerebral cortex present as drug resistant seizure disorder, those involving cerebellum present as mass lesions or slowly progressing vertigo. We report 17 cases, representing the histological spectrum of dysplastic, glioneuronal migration disorders which include, hemimegalencephaly (1), tuberous sclerosis (4), Sturge Weber Syndrome with focal dysplasia (1), Dysembryoplastic neuroepithelial tumor (7) and Lhermitte Ductos disease of cerebellum (2). The dysplastic neurons in varied stages of maturation showed neuronal cytoskeletal pathology similar to that in neuro degenerative diseases, especially when associated with cytomegaly. Similarly, cells exhibiting dual expression of glial and neuronal markers were noted in the cerebral dysplastic lesions. The dysplastic glial elements probably form the subependymal giant cell astrocytomas. Dysplastic neuronal elements form the nidus for DNT. When localized, surgical resection ameliorate the symptoms in many of these condition. Study of these conditions provide better insight into glioneuronal interaction and maturation of the brain. PMID- 12577094 TI - Hypothalamic digoxin and regulation of body mass index. AB - The hypothalamus produces digoxin, an endogenous membrane Na+-K+ ATPase inhibitor and regulator of neurotransmission. Digoxin being a steroidal glycoside, is synthesised by the isoprenoid pathway. In view of the reports of elevated digoxin levels in metabolic syndrome X with high body mass index, the isoprenoid pathway mediated biochemical cascade was assessed in individuals with high and low body mass index. It was also assessed in individuals with differing hemispheric dominance to find out the relationship between digoxin status, body mass index and hemispheric dominance. The isoprenoid pathway metabolites, tryptophan / tyrosine catabolic patterns and membrane composition were assessed. In individuals with high body mass index an upregulated isoprenoid pathway with increased HMG CoA reductase activity, serum digoxin and dolichol levels and low ubiquinone levels were observed. The RBC membrane Na+-K+ ATPase activity and serum magnesium levels were decreased. The tyrosine catabolites (dopamine, morphine, epinephrine and norepinephrine) were reduced and the tryptophan catabolites (serotonin, quinolinic acid, strychnine and nicotine) were increased. There was an increase in membrane cholesterol : phospholipid ratio and a reduction in membrane glycoconjugates in individuals with high body mass index. The reverse patterns were seen in individuals with low body mass index. The patterns in individuals with high body mass index and low body mass index correlated with right hemispheric dominance and left hemispheric dominance respectively. Hemispheric dominance and digoxin status regulates the differential metabolic pattern observed in individuals with high and low body mass index. PMID- 12577095 TI - Assessment of cerebral blood flow velocities in pre and post angiographic states with transcranial doppler. AB - The frequency of complications resulting from angiograms reported in the literature vary between 0.2-5 percent. This study was planned to determine the changes in cerebral blood flow velocity before and after angiography, using transcranial doppler in patients of subarachnoid hemorrhage (SAH) undergoing angiographies. Thirty patients with subarachnoid hemorrhage underwent transcranial doppler ultrasonography immediately before and after angiography. Nonionic water-soluble agents were used during the angiograms. The mean flow velocity (MFV) and pulsatility index (PI) at the M1 segment of both middle cerebral arteries was simultaneously measured. When the patients (11 male, 19 female, mean age+SD; 52.45+12.06) were compared according to changes in MFV and PI, pre and post-angiography, there was no statistical difference in MFV (p=0.51 and p=0.99, left and right side respectively), and in PI (p=0.48 and p=0.66) pre and post angiography. Although angiogram can be used to detect vasospasm in SAH, it can also be cause of vasospasm, partially due to the effect of the contrast agent on the cerebral arteries. This study proposes that the angiographic method is still safe and TCD can be used to follow up any possible changes in diameter of cerebral arteries before and after angiography. PMID- 12577096 TI - Oligodendroglioma: clinical profile and treatment results. AB - Oligodendrogliomas are rare glial tumors with a long natural history. Radical surgery, postoperative radiotherapy, chemotherapy and observation have been suggested as part of primary management. This series reports the presentation and outcome in 34 patients with histologically verified pure oligodendrogliomas treated between 1983 and 1993. All patients were treated at the time of initial diagnosis with surgery, with or without additional radiotherapy. The overall survival and progression free survival rates at five years were 69.8% and 55% respectively. There was no tumor or treatment related factor influencing survival significantly. PMID- 12577097 TI - Pitfalls in diagnosis of epilepsy of Janz and its implications. AB - 84 patients of juvenile myoclonic epilepsy (JME) of Janz were studied. Diagnosis was confirmed using clinical and electro-encephalographic (EEG) criterias. 58 (78%) patients of JME were referred as 'refractory or uncontrolled seizures'. Ignoring myoclonic episodes and non-use of activation procedures in EEG were important reasons for diagnostic delay. Sodium valproate (VPA) or clonazepam are the drugs of choice while phenobarbitone (PB), carbamazepine (CZ), and phenytoin (PHT) are ineffective. Clinical spectrum of JME is slightly different in India. Family history of epilepsy or JME is not forthcoming and there is gross delay in the diagnosis. Other differences include age of presentation and mild cognitive impairment. All juvenile patients of generalized epilepsy, not responding to more commonly used CZ, PB and PHT should be strongly suspected for JME by carefully searching for myoclonus. PMID- 12577098 TI - Naegleria meningitis: a rare survival. AB - Acute amebic meningoencephalitis caused by free-living amebae naegleria fowleri is extremely rare and uniformly fatal with only seven survivals reported till date. An interesting case of naegleria meningitis diagnosed by wet mount cytology of cerebrospinal fluid (CSF) and treated with amphoterecin B, rifampicin and ornidazole with complete recovery is presented. In cases of suspected pyogenic meningitis, if CSF staining, antigen detection or culture is negative for bacteria, a wet mount cytology of CSF for naegleria is suggested. Early treatment with amphoterecin B and rifampicin may improve survival. PMID- 12577099 TI - Lithium - induced tardive dystonia. AB - Tardive dystonia is an uncommon form of chronic dystonia, which usually develops on exposure to neuroleptics. Tardive dystonia (Tdt) following lithium therapy has not been previously reported. The case of 38 year old man with bipolar affective disorder who developed tardive dystonia while on maintenance lithium treatment is described. Presentation of Tdt in this patient was fairly characteristic although there was no suggestion of recent neuroleptic exposure. Tdt known to have poor treatment response, responded very well to clozapine, a novel anti-psychotic, in this case. To conclude, Tdt may develop on exposure to drugs other than neuroleptics. An adequate trial to clozapine can prove to be a useful treatment option. PMID- 12577100 TI - Endolymphatic sac tumor: a rare cerebellopontine angle tumor. AB - Endolymphatic sac tumors (ELST) are rare papillary tumors of the temporal bone. Previously named as aggressive papillary middle ear tumors, they have recently been shown to arise from the endolymphatic sac. They are a rare in cerebello pontine angle (CPA). We present a case of an ELST who presented as a CPA tumor with hydrocephalus. He underwent a ventriculo-peritoneal shunt initially. On exploration of the CP angle, the tumor was found to be extremely vascular. He was re-explored following embolization, and a subtotal excision of the tumor was done. Extensive petrous bone infiltration and vascularity of the tumor makes total excision almost impossible with high risk of cranial nerve deficits, excessive blood loss and CSF leak. This tumor should be considered in the differential diagnosis of vascular CPA tumors which erode the petrous temporal bone. The relevant literature is reviewed. PMID- 12577101 TI - Asymptomatic spontaneous rupture of suprasellar dermoid cyst: a case report. AB - Suprasellar dermoid cysts are uncommon intracranial lesions. CT and MRI findings in a rare case of asymptomatic rupture of suprasellar dermoid cyst with subarachnoid dissemination is described. PMID- 12577102 TI - Acanthameba meningoencephalitis: a case report. AB - A 45 year old lady presented with history of recent surgery for uterovaginal prolapse and retained vaginal tampons following which she developed chronic meningitis due to acanthameba infection. Patient responded to a regimen containing albendazole. She was left with hydrocephalus as a sequelae. PMID- 12577103 TI - Bereitschaftspotential in tic disorders: a preliminary observation. AB - Sensory phenomena in tic disorders such as Tourette's syndrome are known but are substantiated by only a handful of studies. In this preliminary report, we studied premonitory urge, a type of sensory phenomenon in three patients of tic disorders. Bereitschaftspotential, a movement-related cognitive potential indicative of motor preparation, was assessed in these patients. As bereitschaftspotential was observed in all our cases prior to occurrence of tics, it is speculated that tics are not entirely involuntary but are quasi-volitional in nature. Bereitschaftspotential may thus represent a neurophysiological marker of premonitory urge in tic disorders. Implications of exploring the voluntary nature of tics are discussed. PMID- 12577104 TI - Hemorrhagic necrosis of pituitary adenomas. AB - A clinicopathological study of 41 cases of pituitary apoplexy in a series of 324 surgically treated pituitary adenomas is presented. In 23 patients, the predominant operative finding was hemorrhage with or without necrosis. However, there were 15 (37.7%) cases where pale, necrotic tissue with no evidence of hemorrhage was found at surgery. Pale, necrotic material was particularly found when there was a long interval between the acute clinical event and surgery. It is concluded that the pale, necrotic debris represents one stage in the resorption process of blood after hemorrhagic necrosis of pituitary adenomas. This entity needs to be kept in mind especially since the material closely resemble the pultaceous material seen in craniopharyngiomas and epidermoid cysts. PMID- 12577105 TI - Spinal intramedullary tuberculoma and abscess: a rare cause of paraparesis. AB - Five cases of spinal intramedullary tuberculomas (IMT) and one case of spinal intramedullary tuberculous abscess (ITA) are presented. Gd enhanced MRI revealed ring enhancing lesion with central hypodensity, suggesting granulomatous pathology. Surgical excision of the intramedullary lesions was carried out in four cases, while two patients received presumptive anti-tuberculous chemotherapy only. Repeat MRI after completion of anti-tuberculous therapy showed total resolution of the lesion. In other cases following surgical excision, the patients improved significantly. The management of these rare lesions is discussed and the literature reviewed. PMID- 12577106 TI - Tuberculoma en plaque: a case report. AB - A 14 year old girl presented with a 6-month history of headaches and visual blurring. CT showed a right tentorial enhancing lesion. Angiogram showed no enhancement. Histopathological examination of the excised material was consistent with tuberculoma. The presentation of such unusual intracranial tuberculoma is reported and the literature reviewed on the subject. PMID- 12577107 TI - Autosomal recessive limb girdle myasthenia in two sisters. AB - Limb girdle myasthenic syndromes are rare genetic disorders described under the broad heterogeneous group known as congenital myasthenic syndromes and present with mixed features of myasthenia and myopathy. The familial limb girdle myasthenia has been described as one with selective weakness of pectoral and pelvic girdles, showing a positive response to edrophonium chloride. A report of two sisters affected by this disorder is presented. PMID- 12577108 TI - Scleroderma, stroke, optic neuropathy: a rare association. AB - A known case of scleroderma presented with right hemiparesis, focal seizures, optic atrophy and gangrene of digits. There was no evidence of peripheral nerve or muscle involvement. MRI showed multifocal infarcts in both cerebral hemispheres. MR angiography revealed poor flow in bilateral carotid arteries with collateralization from posterior circulation. She improved with phenytoin, nifedipine, antibiotics and immunosuppressants. The rarity of central nervous system affliction in scleroderma and large vessel vasculitis is discussed along with review of literature. PMID- 12577109 TI - Subarachnoid hemorrhage and intracereebral hematoma following lumboperitoneal shunt for pseudotumor cerebri: a rare complication. AB - Placement of lumboperitoneal (LP) shunt as a surgical treatment for benign intracranial hypertension (BIH) is generally a safe procedure, with complications like mechanical failure, overdrainage and infections. Subarachnoid hemorrhage and intracerebral hematoma were seen after lumboperitoneal shunt in a patient having BIH. These complications were the cause of the patient's deterioration. After removal of the hematoma and performing a decompressive procedure, patient's neurological condition improved. The clinical features, investigations and clinical course are described and the literature reviewed. PMID- 12577110 TI - Aneurysmal bone cyst of the temporal bone: a case report. AB - Aneurysmal bone cyst in temporal region is rare. It is a benign condition and may extend intracranially. Total surgical removal is recommended, whenever possible in one stage or in multiple stages. Prognosis is excellent and total removal will effect a cure. An unusual case of aneurysmal bone cyst of temporal region is reported. PMID- 12577111 TI - Sacral perineural cyst presenting as chronic perineal pain: a case report. AB - We present an interesting case of sacral perineural cyst which caused chronic perineal pain. Perineural cyst is relatively rare, especially the sacral region. Chronic perineural pain is an often encountered problem that is difficult to evaluate and sacral perineural cyst may be the etiology of chronic perineal pain in many instances. PMID- 12577113 TI - Evaluation of enzymes in serum and cerebrospinal fluid in cases of stroke. AB - Levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and creatine kinase (CK) were estimated in serum and cerebrospinal fluid of 25 patients of stroke, and were correlated with severity of disease. 21 (84%) patients had ischemic stroke and four (16%) had hemorrhagic stroke. Serum and CSF AST levels were significantly elevated in the study group. The rise in CSF AST was more in the hemorrhagic subtype than in the ischemic subtype. Serum ALT and CSF LDH levels were also significantly elevated in patients with ischemic stroke. None of the enzyme levels were related to the severity of disease as assessed by the Glasgow coma scale. PMID- 12577112 TI - Diffuse neurofibroma of scalp. AB - Diffuse neurofibroma is an uncommon but distinct variety of neurofibroma, usually affecting trunk, head and neck regions of adolescents and young adults. The clinical features, gross macroscopic and histopathological findings are enunciated and the criteria for instituting the preferred modality of treatment for such lesions has been reviewed, stressing upon the need to exclude the neurofibromatoses preoperatively. PMID- 12577114 TI - A patient with optic pathway glioma, scoliosis, Chiari type I malformation and syringomyelia: is it Neurofibromatosis type 1? AB - A 22 years old girl had features of optic pathway glioma, scoliosis, Chiari type 1 malformation and cervical syringomyelia. She had no cutaneous lesions. We considered this combination to be more than coincidental and argue in favour of considering the case as a variant form of Neurofibromatosis type 1. The relevent literature in favour of our contention has been reviewed. PMID- 12577115 TI - A case of hemeplegic migraine with leptomeningeal angiomatosis. AB - A 43 year old man with thirty years history of recurrent hemiplegic migraine, consistently occurring on one side (left sided paresthesia and weakness of less than forty five minutes followed by right sided headache) and lately increasing to once in three days was investigated. CT head and carotid angiogram showed gyral calcification with prominent medullary and subependymal veins confirming the diagnosis of leptomeningeal angiomatosis. Management and followup is discussed. PMID- 12577116 TI - Finger tapping activates spikes in benign epilepsy with centro-temporal spikes. AB - A case of benign epilepsy with centro-temporal spikes (BECT) is reported, in whom tapping of fingures activated typical spikes. PMID- 12577117 TI - Circadian pattern in cerebro vascular disorders. AB - Over the last decade, various studies have been reported to evaluate the circadian pattern of cardiovascular and cerebro-vascular diseases. The data from Indian population is lacking. We undertook this prospective observational study to evaluate the circadian variation in disorders like cerebro-vascular accidents and transient ischemic attacks. Total of 146 patients (events) were studied. Only 10 patients had TIA's. 55% had hemorrhage and 45% had infarction. The 24 hours period was divided into 6 equal portions of 4 hours each. The maximum events were seen between 4 am to 8 am and 12 noon to 4 pm (23.28%) each. Minimum events were seen between 12 midnight to 4 am 14/146 - 9.58%). The circadian variation in occurrence of cerebro-vascular disorders was present with two equal peaks. PMID- 12577118 TI - Visual loss with papilledema in Guillain-Barre syndrome. AB - Papilledema and raised intracranial pressure have been reported in association with Guillain-Barre syndrome. Papilledema is usually asympotomatic or associated with mild visual field defects, without any visual loss. The cerebrospinal fluid protein is usually reported to be high. A case of a 35 year old lady is reported, who presented with headache, diplopia and progressive visual loss in both eyes and limb weakness with hyporeflexia. Optic fundus examination showed bilateral papilledema. She had features of pseudotumor cerebri. Nerve conduction studies were suggestive of polyradiculopathy. The unusual things in this case, were the profound visual loss normal cerebrospinal fluid profiles and the presentation of papilledema before the limb weakness. PMID- 12577119 TI - Spinal subdural epidermoids - a separate entity: report of 3 cases. AB - Intradural extramedullary epidermoid cysts are rare tumors especially those not associated with spinal dysraphism. We report 3 cases of spinal intradural extramedullary epidermoid cysts. In all the cases, the lesion was situated at dorsal level. The clinical features, MRI characteristics and surgical treatment of such rare intradural extramedullary benign tumors are discussed and relevant literature is reviewed. PMID- 12577120 TI - The effects of cold stress test on vasomotor tonus in normal controls. AB - Plethsymographic recordings are used to evaluate changes in peripheral vascular tonus. Twenty-six volunteers (15 men, 11 women) with a mean age 31 years were included in this study. Plethsymographic recordings were done both at baseline and +40C temperatures bilaterally. Significant reductions in systemic blood flow were noted with different maneuvers. Reactive hyperemia occurring after ischemia was local, however local reduction in blood flow after cold test was systemic. As the local temperature increased, the blood flow parameters normalized, too. In normal controls these values normalized in 20 minutes. The changes in vasomotor tonus due to cold test demostrated a parallel pattern. PMID- 12577121 TI - Skull base chondroblastoma: a case report. AB - Chondroblastoma is a rare tumor of the skull. Temporal bone is the commonest site of involvement in the skull. We present a thirty one year old man who presented with painless swelling over the left temporal bone, which was near totally excised after preoperative embolization. Management of this unusual tumor and its complications are discussed. PMID- 12577122 TI - Detection of 1583 bp gene transcript in lymphocytes of muscular dystrophy patients. AB - DMD and BMD are X-lined recessive disorders. RAP-PCR was utilized to investigate differentially expressed gene transcripts in lymphocytes from DMD, BMD and normal individuals as possible diagnostic parameter. A 1583 bp transcript was found to be expressed in both DMD and BMD patients which was unrelated to the known dystrophin gene. This may prove helpful in determining the carrier status of DMD/BMD. PMID- 12577123 TI - Images: multiple meningiomas. PMID- 12577124 TI - Spinal meningioma with positive dural tail sign. PMID- 12577125 TI - Neglected maternity: making a difference. PMID- 12577126 TI - Screening for gestational diabetes mellitus. PMID- 12577127 TI - Gonadotropin-releasing hormone and reproductive medicine. AB - The hypothalamic gonadotropin-releasing hormone (GnRH) is a key regulator of the reproductive system, mainly through its effects on pituitary luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release. Gonadotropin-releasing hormone analogues are modifications of the natural decapeptide, being either agonists (GnRHa) or antagonists. GnRHa may imitate the native hormone and induce an endogenous LH surge; however, sustained treatment with GnRHa results in complete refractoriness of LH and FSH production. This aspect of GnRHa action is the main application of the GnRHa in reproduction today. The hypogonadotropic hypogonadal state that results from the sustained treatment is the basis for additional applications of the GnRHa. The agonists appear to be effective, to some extent, for the treatment of endometriosis and were proved to be beneficial prior to surgery for fibroma uteri. GnRHa is used in assisted reproduction for both induction of an endogenous LH surge and for induction of ovulation, and its capacity to cause refractoriness of the pituitary may be used to avoid a premature LH surge. It is suggested that the GnRHa have a direct effect on ovarian steroidogenesis, which is independent of its action on the pituitary. This unwanted effect and other possible drawbacks of the agonists are thought to be eliminated with the use of the antagonists. The mechanism of action of these antagonists is through competitive blocking of the GnRH receptor, which results in a rapid but reversible suppression of gonadotropin secretion. As for GnRHa, GnRH antagonists have been proven effective to prevent the LH surge and to reduce the incidence of severe ovarian hyperstimulation syndrome in controlled ovarian stimulation cycles. PMID- 12577128 TI - The prevention of congenital anomalies with periconceptional folic acid supplementation. AB - Large randomized trials have demonstrated the critical role of folic acid supplementation in the prevention of neural tube defects. Since 1992, a number of national and international professional societies have released guidelines recommending folic acid supplementation of at least 0.4 mg/day for all women of childbearing age or women planning pregnancies, and 4 mg/day for women with a previous infant with a neural tube defect. Furthermore, a reduction in the incidence of congenital cardiac and urologic anomalies has been demonstrated in some studies following periconceptional folic acid supplementation. There is growing evidence of a possible role of folic acid supplementation in the prevention of other congenital anomalies, including cardiac defects. Since 1998, mandatory fortification of certain foods with folate has been associated with at least a 54% reduction in the incidence of open neural tube defects, yet rates of periconceptional folic acid use remain suboptimal, especially in minority women. Innovative strategies in educating women, health-care providers, and pharmacists on the benefits of folic acid supplementation need to be explored. Mandatory folate fortification of foods needs to be defined and monitored. PMID- 12577129 TI - The association of placental abnormalities with maternal and neonatal clinical findings: a retrospective cohort study. AB - OBJECTIVES: (1) To determine the nature and extent of placental pathologic findings; (2) to associate placental pathologic findings with clinical indicators of infection; (3) to evaluate placental pathology in the context of the guidelines outlined by the College of American Pathologists (CAP). METHODS: A retrospective cohort study, through review of maternal and neonatal charts and placental pathology, of 100 sequential pregnancies in which placentas were submitted to pathology. Data were examined using descriptive statistics, and proportional differences were compared using the chi-square test and Fisher's exact test. RESULTS: Overall, 75% of placentas submitted for pathology review had pathologic abnormalities. Fifty percent had findings consistent with inflammation, 38% had findings consistent with vascular abnormalities, and 18% had findings consistent with meconium. Fetal clinical indicators of infection were associated with placental findings of chorioamnionitis (p < or = 0.01), while maternal clinical indicators were not. Similarly, fetal clinical indicators were associated with placental findings of fetal inflammation (p < or = 0.025), whereas maternal indicators were not associated with placental findings of maternal inflammation. A diagnosis of chorioamnionitis in labour by the attending physician was associated with pathologic findings (p < or = 0.05). A CAP indication was found in 75% of the placentas. There was no difference in incidence of placental pathology between those placentas submitted with and without a CAP indication. CONCLUSION: Placental findings of inflammation or infection were associated with fetal clinical indicators of infection, but not with maternal indicators. Placental pathology is very useful in identifying undiagnosed maternal infection or inflammation. PMID- 12577130 TI - Bereavement support for women and their families after stillbirth. AB - OBJECTIVES: (1) To heighten awareness of the grieving process of the mother and her family experiencing the death of a baby; (2) to offer suggestions to health care providers of the type of support that will achieve optimal grief resolution. OPTIONS: Early, late, or no interventions for women and families who experienced stillbirths. OUTCOME: Success of health-care providers in preventing, recognizing, and treating psychological problems in the bereaved parents and families, and also in helping these families to build meaningful experiences and positive memories from their loss. EVIDENCE: English-language articles and their references on grief and bereavement after perinatal death, through a search of MEDLINE, the Cochrane Library, and publications of other national bodies including the Canadian Paediatric Society, and the American College of Obstetricians and Gynecologists. PMID- 12577131 TI - Lying in the bed we've made: reflection on some unintended consequences of clinical practice guidelines in the courts. AB - Evidence on the use of clinical practice guidelines to aid in the legal determination of negligence is increasing, specifically where they affect determinations of the standard of care and causation. So too is evidence that some clinical practice guidelines are of poor quality. An action alleging the negligent failure to diagnose and treat gestational diabetes in 1988, in which the neonate suffered permanent brachial plexus injury, entered into evidence a 1984 clinical practice guideline authored by the Society of Obstetricians and Gynaecologists of Canada. No "experts" were called to adjudicate the quality of this guideline, which cited no evidence or rationale in support of its recommendations. The standard as laid out in the guideline was judged by the court to reflect a prevailing standard of care, and a finding of negligence was rendered. As the courts pay increased attention to clinical practice guidelines, critical appraisal by the professional organizations developing these documents must occur to assure methodological rigour. Further, the quality of clinical practice guidelines should receive critical scrutiny by the courts if they are to be relied upon, even partially, to assist with legal determinations of the standard of care or issues under causation. PMID- 12577132 TI - Amniocentesis and women with hepatitis B, hepatitis C, or human immunodeficiency virus. AB - OBJECTIVE: To review the risk of in utero infection through amniocentesis in women with hepatitis B, hepatitis C, or human immunodeficiency virus (HIV). OUTCOMES: Fetal and neonatal morbidity and mortality. EVIDENCE: Review articles, meta-analyses, and MEDLINE searches from 1966 to 2002 for English-language articles related to amniocentesis, fetal and neonatal infection, and hepatitis B, hepatitis C, or HIV. VALUES: The evidence collected was reviewed by the Genetics Committee of the Society of Obstetricians and Gynaecologists of Canada (SOGC) and quantified using the Evaluation of Evidence guidelines developed by the Canadian Task Force on the Periodic Health Exam. RECOMMENDATIONS: 1. The risk of fetal hepatitis B infection through amniocentesis is low. However, knowledge of the maternal hepatitis B e antigen status is valuable in the counselling of risks associated with amniocentesis. (II-1C) 2. Amniocentesis in women infected with hepatitis C does not appear to significantly increase the risk of vertical transmission, but women should be counselled that very few studies have properly addressed this possibility. (II-2C) 3. In HIV-positive women all noninvasive screening tools should be used prior to considering amniocentesis. (II-2D) 4. For women infected with hepatitis B, hepatitis C, or HIV, the addition of noninvasive methods of prenatal risk screening, such as nuchal translucency, triple screening, and anatomic ultrasound, may help in reducing the age-related risk to a level below the threshold for genetic amniocentesis. (II-2C) 5. For those women infected with hepatitis B, hepatitis C, or HIV who insist on amniocentesis, every effort should be made to avoid inserting the needle through the placenta. (II-1B) VALIDATION: These guidelines have been approved by the SOGC Genetics Committee, SOGC Executive, and SOGC Council. SPONSORS: The Society of Obstetricians and Gynaecologists of Canada. PMID- 12577133 TI - Controversies in the management of type II "branch" endoleaks following endovascular abdominal aortic aneurysm repair. AB - Successful endovascular aortic aneurysm repair (EVAR) is often defined as complete exclusion of blood flow within the aneurysm sac. Perigraft flow, also known as endoleak, is the most common complication following EVAR. Attachment site related endoleaks (type I) are generally considered to warrant some form of intervention due to the belief that they represent a risk for future rupture. Management of type II endoleaks, also known as branch or collateral endoleaks, is more controversial. Some advocate a policy of watchful-waiting whereas others treat all type II endoleaks as soon as they are discovered. The following review explores the controversies pertaining to the management, diagnosis and surveillance imaging, and treatment of type II endoleaks. PMID- 12577134 TI - Validation of two risk models for perioperative mortality in patients undergoing elective abdominal aortic aneurysm surgery. AB - The authors validated 2 clinical risk models for perioperative mortality in patients scheduled for elective open abdominal aortic aneurysm surgery (AAA surgery). They studied 361 patients who underwent elective AAA surgery between 1991 and 1999 (validation population). Two clinical risk models were validated. The first was developed in 238 patients from Leiden University Hospital (Leiden risk model). The Leiden risk model was modified to provide predictions for the validation population based on 6 predictors including age, gender, a history of previous myocardial infarction, congestive heart failure, renal disease, and pulmonary disease. The second was a recently published simpler risk model developed in 820 patients in the UK Small Aneurysm Trial (UK risk model) and included 3 predictors (age and renal and pulmonary comorbidity). Logistic regression was used to quantify the relationship between predictors and outcome (mortality within 30 days of surgery). Validation further included the concordance statistic (c-statistic) for discriminative ability and the Hosmer Lemeshow test for model reliability. The perioperative mortality in the validation population was 6.6% (24/361). Predictors had similar odds ratios, with particularly strong effects of congestive heart failure, pulmonary disease, and renal impairment. The Leiden risk model had reasonable good ability (c-statistic 0.72) and showed adequate calibration (chi(2)=3.3, p=0.97). It could particularly identify a low-risk group. The UK risk model did not perform well (c-statistic 0.60), showing statistically significant lack of fit (chi(2)=64.9, p<0.001). This study showed similar predictive ability of previously identified predictors for perioperative mortality. The Leiden risk model could identify a low-risk group, while the UK risk model showed a relatively poor performance. The current study supports the use of the Leiden model for preoperative risk assessment. PMID- 12577135 TI - Morphometric analysis of the popliteal artery for endovascular treatment. AB - This study intended to determine the precise diameter of the popliteal artery in patients at risk for popliteal aneurysms. Accurate sizing is necessary to develop devices for endovascular treatment of popliteal aneurysms. Fifty-four patients with abdominal aortic aneurysms (AAAs) had computed tomography (CT) scans of the popliteal arteries. Age- and gender-matched control subjects were measured by ultrasound. NIH Image was used to measure the minor diameter at the adductor hiatus (proximal) and femoral condyles (midpopliteal artery). There were 4 unsuspected popliteal aneurysms (7.4%). The proximal popliteal artery was ectatic in these patients: 13.4 +/- 5.2 mm. Proximal and midpopliteal arteries were significantly larger in the other patients with AAAs compared with controls: 9.6 +/- 1.8 mm vs 7.9 +/- 1.1 mm at the hiatus (p<0.001) and 10.2 +/- 2 mm vs 7.9 +/- 0.9 mm at the condyles (p<0.001). The popliteal artery was focally larger in patients with AAAs without popliteal aneurysms. The popliteal artery was larger in men compared with women; 9.8 +/- 1.8 mm vs 8.8 +/- 1.9 mm at the hiatus (p=0.024) and 10.5 +/- 1.9 mm vs 9.0 +/- 2.4 mm at the condyles (p=0.005). The proximal popliteal artery was 2 mm larger in patients at risk for popliteal aneurysms and 5 mm larger in patients with popliteal aneurysms compared to controls. Focal ectasia of the midpopliteal artery was common. Planning for endovascular treatment of popliteal aneurysms must incorporate this striking enlargement. PMID- 12577136 TI - Single-center review of infections associated with 1,574 consecutive vascular access procedures. AB - The objective of this study was to establish accurate infection rates and causes for vascular access procedures and provide information helpful in identifying means of reducing infections. Between January 1, 1996, and December 31, 2001 (60 months), a single surgeon (AFS) performed 1,574 consecutive vascular access procedures (850 patients) in 1 academic medical center. Recently, a retrospective review of these records revealed that 87 patients (10.2% of this population) underwent removal of access grafts or fistula for infection. In these 87 patients, 131 grafts and 1 fistula were removed. Thus, 8.4% (132/1,574) of the total surgeries were for infection. This review included evaluation of repeat infection, causative organism, and HIV status. Cause of infection fell into 5 major categories. Of the 87 patients, 26 had multiple graft infections. Diabetes and hypertension lead to increased end-stage renal disease, requiring more vascular access procedures. Following thrombosis, infection is the most common complication in vascular access. Therefore, mechanisms to reduce infection would be well received. These data suggest that operative infection accounted for only 6% of infective procedures, and when the entire population is considered, the operative infection rate was 0.51%. However, infections secondary to routine dialysis make up more than 50% of this infective operative population. The authors note that 17% of the infective operations were in previously infected grafts that were not initially completely removed and 23% developed in nonfunctional grafts. Multiple graft infections played a significant role in the investigation and warrant further evaluation. PMID- 12577137 TI - Temporary hypotension following endarterectomy for severe carotid stenosis: should we treat it? AB - In patients with severe carotid stenosis, the author has observed that temporary low blood pressure often occurs in the postoperative period. The hypotension typically develops 2 to 4 hours following operation, is asymptomatic, and resolves within 12-24 hours. In recent years treatment has consisted of simple observation, avoiding the use of vasopressors. Other reports of hemodynamic instability following carotid endarterectomy emphasize high blood pressure. The author rarely observes postoperative hypertension in his practice and has wondered if technical factors explain the difference. A consecutive series of primary carotid endarterectomies recently performed by the author was prospectively studied to determine the incidence of postoperative hypotension. Forty-nine of 180 consecutive patients (27%) developed hypotension below 90 mm Hg systolic (range 65-90). All had severe stenosis as an indication for operation. Only 1 patient required treatment for symptoms related to the low blood pressure. Simple observation of the remaining 48 patients with postoperative hypotension did not result in complications nor delay discharge. Data management, selection of patients, surgical technique, and results are discussed. Temporary postoperative hypotension in patients with severe carotid stenosis is common and appears to be benign. The author speculates that this phenomenon may protect ischemic brain tissue from sudden hyperperfusion, and as such should not be treated with vasopressor medication unless symptoms are present. PMID- 12577138 TI - Review of standards for competence in catheter-based endovascular procedures: a resource and strategy for the interventional vascular surgeon. AB - The field of peripheral vascular disease management is evolving. As such, vascular surgeons can no longer be complacent and expect to be included in the rapidly expanding practice of endovascular intervention. If not engaged, vascular surgeons risk losing not only the patients who have largely driven this evolution but also their role as leaders in the management of vascular disease. Therefore vascular surgeons and vascular surgical training programs find themselves in an awkward and sometimes confrontational circumstance attempting to gain experience and training in catheter-based procedures. Individual societies, both surgical and nonsurgical, have put forth competence standards for catheter-based procedures. However these standards are not always consistent and the definition of competence is often a heated debate. In addition, combining the acquisition of endovascular skills with the multitude of available endovascular training experiences, didactic and "hands-on," is not well coordinated. The objectives of this review are to summarize the literature as it relates to catheter-based endovascular competence, including publications from nonsurgical catheter-based specialties, and to place the acquisition of basic endovascular skills in context with the available endovascular courses in a stepwise strategy. A final objective is to provide a literature-based resource that outlines specific phases in the development of interventional vascular surgeons and training programs as they advance into the arena of catheter-based endovascular therapies. PMID- 12577139 TI - Arterial wall shear stress: observations from the bench to the bedside. AB - Shear stress is the tangential force of the flowing blood on the endothelial surface of the blood vessel. Shear is described mathematically or ideal fluids, and in vitro models have enabled researchers to describe the effects of shear on endothelial cells. High shear stress, as found in laminar flow, promotes endothelial cell survival and quiescence, alignment in the direction of flow, and secretion of substances that promote vasodilation and anticoagulation. Low shear stress, or changing shear stress direction as found in turbulent flow, promotes endothelial proliferation and apoptosis, shape change, and secretion of substances that promote vasoconstriction, coagulation, and platelet aggregation. The precise pathways by which endothelial cells sense shear stress to promote their quiescent or activated pathways are currently unknown. Clinical applications include increasing shear stress via creation of an arteriovenous fistula or vein cuff to promote bypass graft flow and patency. Since an abnormal level of shear stress is implicated in the pathogenesis of atherosclerosis, neointimal hyperplasia, and aneurysmal disease, additional research to understand the effects of shear stress on the blood vessel may provide insight to prevent vascular disease. PMID- 12577140 TI - Diagnosis and management of aneurysms involving the superior mesenteric artery and its branches--a report of four cases. AB - Aneurysms of the superior mesenteric artery (SMA) are an uncommon but lethal entity, which must be treated expeditiously to avoid mortality and high incidence of ischemic small bowel complications. In the past 7 years the authors have treated 4 patients with a variety of types of aneurysms involving the SMA and its branches at a university-based teaching hospital. The first was a mycotic SMA aneurysm as a result of septic mitral valve, the second a jejunal aneurysm in a patient with pancreatitis, the third a spontaneous dissection distal to a small SMA aneurysm with thrombus partially occluding the distal vessel, and the fourth an SMA aneurysm associated with the diagnosis of mesenteric insufficiency. All patients presented with abdominal pain. The diagnosis was made initially in 1 patient on plain abdominal films with a calcified aneurysm, on duplex scan in the second, and on computed tomography (CT) scans in the remaining 2. Treatment consisted of bowel resection and ligation of mycotic aneurysm in the first patient, of catheter embolization of jejunal aneurysm in the patient with pancreatitis, and of vein graft bypass in the patient with a large SMA aneurysm. The patient with SMA aneurysm and distal dissection with partially occluding thrombus received anticoagulation and is being followed up with serial CT scans. There were no deaths. One patient required bowel resection, which did not result in short gut syndrome. Improved abdominal duplex scanning and CT technology facilitates the diagnosis of mesenteric aneurysm. The broad spectrum of etiologies mandates that treatment be tailored to the individual patient, and it varies from endovascular techniques to traditional bypass surgery. Prompt diagnosis and treatment results in the lowest mortality rate and minimizes the prevalence of intestinal infarction. PMID- 12577141 TI - Endovascular repair of an arteriovenous fistula from a ruptured hypogastric artery aneurysm--a case report. AB - A spontaneous ilioiliac arteriovenous fistula secondary to rupture of a hypogastric artery aneurysm is an unusual occurrence. A case of an endovascular repair of this challenging problem is reported. PMID- 12577142 TI - Acute thrombosis of abdominal aortic aneurysms--report of two cases and review of the literature. AB - An acutely thrombosed abdominal aortic aneurysm (AAA) is a rare but devastating complication of aortic aneurysms. Incidence in reported series is approximately 0.7-2.8% of surgically managed AAA cases with an associated mortality rate of 50%. To date there have been only 46 cases of acutely occluded AAAs reported in the literature. We present our experience of 2 additional cases of acute AAA thrombosis and discuss the mechanism of thrombosis and management. The size of the aneurysm seems to be a better predictor of rupture than of thrombosis. Strong risk factors for thrombosis are the presence of concomitant iliac artery and cardiac disease. Although this complication is rare, its high mortality rate makes it an important disease to understand, recognize, diagnose, and treat for a successful outcome. PMID- 12577143 TI - Regarding "Vascular surgeon Edwin J. Wylie, MD (1918-1982), pioneer and visionary". PMID- 12577144 TI - Continuous is not continuous: the incidence and impact of circuit "down-time" on uraemic control during continuous veno-venous haemofiltration. AB - OBJECTIVE: There is little information on the duration of time that patients spend off therapy (down-time) during continuous veno-venous haemofiltration (CVVH) and the effect of this treatment free time on azotaemic control. DESIGN AND SETTING: Prospective observational study in the ICU of tertiary hospital. PATIENTS AND PARTICIPANTS: 48 critically ill patients treated with CVVH at 2 l/h of ultrafiltration. INTERVENTIONS: Prospective collection of demographic and biochemical data. MEASUREMENTS AND RESULTS: Two hundred and sixty-six filters were observed. Start and end times were collected for each filter. Creatinine and urea were measured daily and percentage of reduction of these two solutes was calculated (%Delta creatinine and urea). The median period when CVVH was not applied to a patient (down-time) was 3 h per day. There was a significant inverse correlation between down-time and %Delta creatinine and urea over each 24-h time cycle. On average at least 16 h per day of CVVH was required to maintain creatinine and urea concentration for each 24-h cycle. CONCLUSIONS: "Continuous" therapy is not truly continuous. Down-time adversely affects azotaemic control. Physicians prescribing CRRT should be aware of the consequences of such down-time on the quality and quantity of renal replacement therapy delivered. PMID- 12577145 TI - Somatosensory evoked potentials as predictor of systemic inflammatory response syndrome in pigs? AB - OBJECTIVE: Sepsis or systemic inflammatory response syndrome (SIRS) is often associated with encephalopathy (70%), which has been described as an early symptom resulting in several diseases. The present study investigated somatosensory evoked potentials (SEP) as an indicator or even a predictor of cerebral dysfunction evaluated in an experimental model of SIRS in pigs. METHODS: Eight Gottinger minipigs were included in the study. SIRS was mediated by induction of pancreatitis due to injection (ductus pancreaticus) of 500 mg/kg sodium taurocholate and 2.5 IU/kg enterokinase. Monitored parameters were: arterial blood-central venous-pulmonary arterial pressure, and cardiac output, systemic vascular resistance, and body temperature. SEP were recorded from centroparietal vs. frontal areas after electrical stimulation of the right forepaw. RESULTS: At least 33% loss of vascular resistance from baseline (SIRS criteria) occurred in all animals within 4-18 h. Baseline recordings in all anesthetized animals indicated primary cortical responses to electrical stimuli identified by peak latencies between 15-20 ms (SEP(P15-20)). Attenuations in the amplitudes with significant median decreases of 46% were observed at least 4 h before the defined hemodynamic SIRS criteria. CONCLUSIONS: The present data show a trend for the attenuation in SEP amplitudes as an indicator of systemic inflammatory response. SEP monitoring may be a sensitive marker of developing early changes in cerebral function due to SIRS-related encephalopathy. PMID- 12577146 TI - Arterial pressure monitoring in septic shock. PMID- 12577147 TI - Impact of pulmonary artery occlusion pressure value on the definition of acute respiratory distress syndrome. PMID- 12577148 TI - Nosocomial bacteremia involving Acinetobacter baumannii in critically ill patients: a matched cohort study. AB - OBJECTIVE: To determine outcome and attributable mortality in critically ill patients with nosocomial bacteremia involving A. baumannii. DESIGN: A retrospective matched cohort study in which all ICU patients with microbiologically documented A. baumannii bacteremia were defined as cases. Matching of the controls was based on equivalent APACHE II score (+/-2 points) and diagnostic category. Control patients were required to have an ICU stay equivalent to or longer than the case prior to onset of the bacteremia. SETTING: The 54-bed ICU of the 1060-bed Ghent University Hospital. PATIENTS: 45 ICU patients with A. baumannii bacteremia and 90 matched control subjects without clinical or microbiological evidence of blood stream infection. MEASUREMENTS: Population characteristics and in-hospital mortality rates of patients with A. baumannii bacteremia and their controls were compared. Attributable mortality is determined by subtracting the crude mortality rate of the controls from the crude mortality rate of the cases. RESULTS: Patients with A. baumannii bacteremia had significantly more hemodynamic instability, longer ICU stay, and longer length of ventilator dependence than controls. In-hospital mortality rates for cases and controls were, respectively, 42.2% and 34.4%; thus the attributable mortality was 7.8%. CONCLUSION: In critically ill patients A. baumannii bacteremia is not associated with a significantly increased mortality rate. PMID- 12577149 TI - Onset of acute respiratory distress syndrome following severe pulmonary hemorrhage in a patient with anti-neutrophil cytoplasmic antibody associated vasculitis. PMID- 12577150 TI - The relationship between methodological trial quality and the effects of impregnated central venous catheters. AB - OBJECTIVE: We assessed the methodological trial quality of individual randomized controlled studies on chlorhexidine silver sulfadiazine impregnated catheters and the effect on catheter-related infection (CRI). DESIGN: Only the studies identified in the Medline database from 1966 to December 2001 were considered, abstracts being excluded. The outcome of this investigation centered on bloodstream CRI. The trial quality of the 11 studies identified and published (total 3,131 catheters) was assessed using a scoring system based on allocation, patient selection, patient characteristics, blinding of the intervention and the diagnosis of CRI (range 0-2 points, 10 points maximum). RESULTS: The mean methodological quality score was 7.1 (range 5-9). The relative risk reduction for bloodstream CRI ranged from -0.14 to 1.0. No association between trial quality and the impact of chlorhexidine silver sulfadiazine impregnated catheters on CRI was found. The summary odds ratio for CRI was 0.69 (95% CI 0.46-1.03). Taking only those studies with 2 points for diagnosis of CRI a summary odds ratio of 0.87 (95% CI 0.44-1.72) resulted, whereas studies with a diagnosis score of only 1 point led to a summary odds ratio of 0.60 (95% CI 0.35-1.02). CONCLUSIONS: The quality of the studies seems to have had no influence on the outcome, according to the results of this investigation, but the use of only a single quality score may not be sufficient to investigate the prevention effect of impregnated catheters. PMID- 12577151 TI - Comparison of effectiveness of two urinary drainage systems in intensive care unit: a prospective, randomized clinical trial. AB - OBJECTIVE: In a previous nonrandomized study we observed no difference in the rate of acquisition of bacteriuria between a complex closed drainage system (CCDS) and a two-chamber drainage system (TCDS) in ICU patients. To confirm this result we performed a statistically powerful study assessing the effectiveness of the CCDS and the TCDS in ICU patients. DESIGN AND SETTING: Randomized, prospective, and controlled study in the medicosurgical intensive care unit (16 beds) in a teaching hospital. PATIENTS AND INTERVENTIONS: We assigned 311 patients requiring indwelling urinary catheter for longer than 48 h to TCDS or CCDS to compare the rate of acquisition of bacteriuria. MEASUREMENTS AND RESULTS: Patients did not receive prophylactic antibiotics during placement management or catheter withdrawal. Urine samples were obtained weekly for the duration of catheterization and within 24 h after catheter removal, and each time symptoms of urinary infection were suspected. There was no statistical difference in the rate of bacteriuria between the two groups: 8% with TCDS and 8.5% with CCDS. Rates of urinary tract infection were 12.1 episodes with TCDS and 12.8 episodes with CCDS per 1000 days of catheter. CONCLUSIONS: This randomized study on the effectiveness of TCDS and CCDS in ICU patients confirms the findings of our previous study. No differences were noted between the two systems. The higher cost of CCDS is not justified for ICU patients. PMID- 12577152 TI - Ventricular interdependence: how does it impact on hemodynamic evaluation in clinical practice? PMID- 12577153 TI - Partial liquid ventilation: effects of positive end-expiratory pressure on perfluorocarbon evaporation from the lungs of anesthetized dogs. AB - OBJECTIVE: Perfluorocarbons are eliminated during partial liquid ventilation mainly by evaporation via the airways. We examined whether this is affected by the level of end-expiratory airway pressure. DESIGN AND SETTING: Observational cohort animal study in the animal laboratory of a university hospital. SUBJECTS: Five foxhound dogs. INTERVENTIONS: The anesthetized dogs underwent partial liquid ventilation (5 ml/kg perfluorocarbon) at constant respiratory rate (17+/-1 breaths/min) and tidal volume (10 ml/kg). The level of end-expiratory airway pressure was varied repeatedly between 0, 5, and 10 cmH(2)O every 25 min. MEASUREMENTS AND RESULTS: Expired gas was collected in reservoirs to determine evaporative perfluorocarbon loss gravimetrically. Any increase in end-expiratory airway pressure increased while any decrease in end-expiratory airway pressure reduced evaporative perfluorocarbon loss. Mean initial elimination at an end expiratory airway pressure of 5 cmH(2)O was 19.6+/-3.8 microl/kg per minute; this decreased by 28% at an end-expiratory airway pressure of 0 cmH(2)O and increased by 46% at an end-expiratory airway pressure of 10 cmH(2)O. At equal levels of end expiratory airway pressure evaporation decreased linearly over time. CONCLUSIONS: Our results suggest that the level of end-expiratory airway pressure is a determinant of evaporative perfluorocarbon loss and may have relevance for maintenance dosing and instillation intervals during partial liquid ventilation. PMID- 12577154 TI - Sodium crocetinate does not alter gut hypercapnic responses or renal energy stores during transient sub-diaphragmatic ischaemia. AB - OBJECTIVES: To evaluate the protection afforded by trans-sodium crocetinate against dysoxia in an animal model of recurrent sub-diaphragmatic ischaemia. DESIGN: Prospective experimental animal study. SETTING: University research laboratory SUBJECTS: Adult male Sprague-Dawley rats. INTERVENTIONS: Twelve adult male Sprague-Dawley rats (340-510 g) were anaesthetised with sodium pentobarbitone 60 mg/kg i.p. and ventilated with oxygen and isoflurane via tracheostomy. Six 2-min episodes of sub-diaphragmatic hypotension (mean pressure 30 mmHg) were induced using a sling around the proximal aorta. Before the third and sixth episodes, saline 1.5 ml/kg was injected into the aortic cannula. In six rats, this saline contained trans-sodium crocetinate 50 microg/ml. MEASUREMENTS AND MAIN RESULTS: Ileal luminal PCO(2) and distal aortic pressure were monitored continuously. Following ischaemic episodes trans-sodium crocetinate had no discernible effect on either degree of PCO(2) elevation or the time to peak PCO(2). No effects on renal energy charge or nucleotide concentrations were detected. UV-visible spectroscopy of the crocetinate preparation showed that some cis isomer was present. CONCLUSIONS: The findings, although limited to one drug dosage in one animal model, bring into question whether trans-sodium crocetinate affects plasma oxygen diffusivity in vivo. Alternative explanations for the negative findings include a TSC-induced exacerbation of arterio-venous oxygen shunting, the brevity of the dysoxic episodes, and the presence of cis isomer. PMID- 12577155 TI - Patient recollection of airway suctioning in the ICU: routine versus a minimally invasive procedure. AB - OBJECTIVE: Many patients have an unpleasant recollection of routine endotracheal suctioning after discharge from the Intensive Care Unit (ICU). We hypothesized that through minimally invasive airway suctioning discomfort and stress may be prevented, resulting in less recollection. DESIGN: A prospective randomized clinical trial. SETTING: Two ICUs at the University Hospital of Groningen, the Netherlands. PATIENTS AND PARTICIPANTS: Adult patients with an intubation period exceeding 24 h were included. INTERVENTIONS: Patients received either routine endotracheal suctioning (RES) or minimally invasive airway suctioning (MIAS) during the duration of intubation. MEASUREMENTS AND RESULTS: Within 3 days after ICU discharge all patients were interviewed, regarding recollection and discomfort of suctioning. The level of discomfort was quantified on a visual analogue scale (VAS). We analyzed data from 208 patients (RES: n=113, and MIAS: n=95). A significantly lower prevalence of recollection of airway suctioning was found in the MIAS group (20%) compared to the RES group (41%) (P-value =0.001). No significant difference in level of discomfort was found between the RES and the MIAS group (P-value =0.136). CONCLUSIONS: Minimally invasive airway suctioning results in a lower prevalence of recollection of airway suction than in RES, but not in discomfort. PMID- 12577156 TI - Endotracheal suctioning versus minimally invasive airway suctioning in intubated patients: a prospective randomised controlled trial. AB - STUDY OBJECTIVE: Endotracheal suctioning in intubated patients is routinely applied in most ICUs but may have negative side effects. We hypothesised that on demand minimally invasive suctioning would have fewer side effects than routine deep endotracheal suctioning, and would be comparable in duration of intubation, length of stay in the ICU, and ICU mortality. DESIGN: Randomised prospective clinical trial. SETTING: In two ICUs at University Hospital Groningen, the Netherlands. PATIENTS: Three hundred and eighty-three patients requiring endotracheal intubation for more than 24 h. INTERVENTIONS: Routine endotracheal suctioning (n=197) using a 49-cm suction catheter was compared with on-demand minimally invasive airway suctioning (n=186) using a suction catheter only 29 cm long. MEASUREMENTS AND RESULTS: No differences were found between the routine endotracheal suctioning group and the minimally invasive airway suctioning group in duration of intubation [median (range) 4 (1-75) versus 5 (1-101) days], ICU stay [median (range) 8 (1-133) versus 7 (1-221) days], ICU mortality (15% versus 17%), and incidence of pulmonary infections (14% versus 13%). Suction-related adverse events occurred more frequently with RES interventions than with MIAS interventions; decreased saturation: 2.7% versus 2.0% (P=0.010); increased systolic blood pressure 24.5% versus 16.8% (P<0.001); increased pulse pressure rate 1.4% versus 0.9% (P=0.007); blood in mucus 3.3% versus 0.9% (P<0.001). CONCLUSIONS: This study demonstrated that minimally invasive airway suctioning in intubated ICU-patients had fewer side effects than routine deep endotracheal suctioning, without being inferior in terms of duration on intubation, length of stay, and mortality. PMID- 12577157 TI - Temperature measurement in intensive care patients: comparison of urinary bladder, oesophageal, rectal, axillary, and inguinal methods versus pulmonary artery core method. AB - OBJECTIVE: Comparisons of urinary bladder, oesophageal, rectal, axillary, and inguinal temperatures versus pulmonary artery temperature. DESIGN: Prospective cohort study. SETTING: Intensive Care Unit of a University-Hospital. PATIENTS: Forty-two intensive care patients requiring a pulmonary artery catheter (PAC). INTERVENTION: Patients requiring PAC and without oesophageal, urinary bladder, and/or rectal disease or recent surgery were included in the study. Temperature was simultaneously monitored with PAC, urinary, oesophageal, and rectal electronic thermometers and with axillary and inguinal gallium-in-glass thermometers. Comparisons used a Bland and Altman method. MEASUREMENTS AND MAIN RESULTS: The pulmonary arterial temperature ranged from 33.7 degrees C to 40.2 degrees C. Urinary bladder temperature was assessed in the last 22 patients. A total of 529 temperature measurement comparisons were carried out (252 comparisons of esophageal, rectal, inguinal, axillary, and pulmonary artery temperature measurements in the first 20 patients, and 277 comparisons with overall methods in the last patients). Nine to 18 temperature measurement comparisons were carried out per patient (median = 13). The mean differences between pulmonary artery temperatures and those of the different methods studied were: oesophageal (0.11+/-0.30 degrees C), rectal (-0.07+/-0.40 degrees C), axillary (0.27+/-0.45 degrees C), inguinal (0.17+/-0.48 degrees C), urinary bladder (-0.21+/-0.20 degrees C). CONCLUSION: In critically ill patients, urinary bladder and oesophageal electronic thermometers are more reliable than the electronic rectal thermometer which is better than inguinal and axillary gallium in-glass thermometers to measure core temperature. PMID- 12577158 TI - Comparison of gravimetric and a double-indicator dilution technique for assessment of extra-vascular lung water in endotoxaemia. AB - OBJECTIVE: To compare a molecular double-indicator dilution technique with the gravimetrical reference method for measurement of extra-vascular lung water in porcine endotoxin shock. DESIGN: Open comparative experimental study. SETTING: Animal research laboratory. MEASUREMENTS AND RESULTS: In fourteen anaesthetised, mechanically ventilated landrace pigs, central and pulmonary haemodynamics as well as pulmonary gas exchange were measured. Extra-vascular lung water was quantitated gravimetrically as well as with a molecular double indicator dilution technique. Eight of these animals were subjected to endotoxaemia, the rest serving as sham controls. No difference in extra-vascular lung water was observed between the two methods in sham animals. Furthermore, extra-vascular lung water assessed with the molecular double-indicator dilution technique at the initiation of endotoxin infusion did not differ significantly from the corresponding values for sham animals. Endotoxaemia induced a hypodynamic shock with concurrent pulmonary hypertension and a pronounced deterioration in gas exchange. No increase in extra-vascular lung water was detected with the molecular double indicator dilution technique in response to endotoxin, whereas this parameter was significantly higher when assessed with the gravimetric method. CONCLUSION: The molecular double-indicator dilution technique showed similar results as the gravimetrical method for assessment of extra-vascular lung water in non endotoxaemic conditions. However, during endotoxin-induced lung injury the molecular double indicator dilution technique failed to detect the significant increase in extra-vascular lung water as measured by the gravimetric method. These data suggest that the molecular double indicator dilution technique may be of limited value during sepsis-induced lung injury. PMID- 12577159 TI - Metabolic changes detected by microdialysis during endotoxin shock and after endotoxin preconditioning. AB - OBJECTIVE: Preconditioning with low doses of endotoxin has been shown to induce endotoxin hyporesponsiveness. The present study was designed to assess the metabolic response of various tissues during endotoxemia and after pretreatment with endotoxin. DESIGN: Controlled experimental animal study. SETTING: Research laboratory of a university hospital. MEASUREMENTS AND RESULTS: Ten pigs were randomly assigned to a control ( n = 5) or a treatment group ( n = 5), the latter receiving incremental doses of endotoxin 5-2 days prior the experiments. Apart from hemodynamics and oxygen transport variables, lactate, glucose, and glycerol were measured in muscle, subcutaneous fat, and hepatic tissue using microdialysis. Endotoxin was infused (1 micro g.kg.h) until the animals died. A significant increase in tissue lactate (eightfold) and glycerol (fivefold) was observed in the control animals. This effect was almost completely abolished in the endotoxin pretreated group. Endotoxin pretreatment had no significant effects on mean arterial pressure [56 (range 34-89) mmHg vs 70 (47-88) mmHg, n.s.] or cardiac output [4.8 (3.0-5.9) l/min vs 3.2 (2.1-4.2) l/min, n.s.], but significantly improved arterial pO(2) and pH ( P<0.05). Increase of oxygen extraction was higher in control animals [from 34% (range 24-47%) to 72% (range 61-79%)] compared to the pretreatment group [from 30% (range 22-42%) to 44% (range 34-50%), P<0.05]. Endotoxin pretreatment increased survival time from 5.3 h (5.0-5.8) to 8.0 h (7.0-8.5) ( P<0.05), respectively. CONCLUSIONS: Microdialysis monitoring revealed that endotoxin preconditioning ameliorates the increase in tissue metabolism during endotoxemia, accompanied by decreased systemic oxygen demand despite unchanged global hemodynamics. PMID- 12577160 TI - [European Association of Urology guidelines on urinary and male genital tract infections]. AB - Today, the classical bacteria that cause venereal diseases, e.g. gonorrhea, syphilis, chancroid and inguinal granuloma, only account for a small proportion of all known sexually transmitted diseases (STDs). Other bacteria and viruses as well as yeasts, protozoa and epizoa must also be regarded as causative organisms of STD. Taken together, all sexually transmitted infections comprise more than 30 relevant STD pathogens. However, not all pathogens that can be sexually transmitted manifest diseases in the genitals and not all infections of the genitals are exclusively sexually transmitted. Concise information and tables summarising the diagnostic and therapeutic management of STDs in the field of urology allow a synoptic overview, and are in agreement with the recent international guidelines of other specialist areas. Special considerations (i.e. HIV infection, pregnancy, infants, allergy) and recommended regimens are presented. PMID- 12577161 TI - [Clinical management of patients with sepsis]. AB - Sepsis and septic shock are the leading causes of death in non-cardiological intensive care units in developed countries despite recent advances in critical care medicine. Sepsis is the systemic inflammatory response to infection, often associated with hypoperfusion followed by tissue injury and organ failure. Activation of monocytes/macrophages and neutrophils with consecutive release of proinflammatory mediators and activation of the coagulation cascade, seem to play a key role in the pathogenesis of sepsis. Elimination of the septic focus,antimicrobial therapy and supportive treatment are the cornerstones of sepsis therapy. Adequate and rapid volume replacement and if necessary application of catecholamines are of highest priority to optimize tissue perfusion. Norepinephrine is the vasopressor of choice and dobutamine the preferred inotropic agent. Most experts recommend hemoglobin levels of 8-10 g/dl in severe sepsis. In addition,lung protective ventilatory strategies as well as enteral and parenteral nutrition are part of the clinical management of septic patients. In mechanically ventilated patients intensive insulin therapy to maintain blood glucose at a level between 80 and 110 mg/dl has significantly reduced mortality.Furthermore,prophylaxis of deep vein thrombosis and of stress ulcer bleeding are individually applied to septic patients. Treatment of septic patients with anti-mediator strategies or high dose AT III were not successful so far. In contrast,now two new promising treatment options may be emerging: application of small doses hydrocortisone and activated protein C [drotrecogin alfa (activated)]. Large and in part multicentric studies especially in the last 2 years now allow the practicing clinician to perform a partially evidence-based management of patients with sepsis. In addition, for the first time two options for specific therapy of sepsis,application of small doses hydrocortisone and activated protein C [drotrecogin alfa (activated)],are available which may further improve prognosis for septic patients. PMID- 12577162 TI - [Anaesthesia for caesarean section. Comparison of two general anaesthetic regimens and spinal anaesthesia]. AB - OBJECTIVES: The study was undertaken to compare different anaesthetic techniques for scheduled or urgent caesarean section with respect to maternal circulatory parameters, maternal and fetal endocrine stress response, fetal vitality parameters and further clinical parameters such as recovery and analgesic demand. METHODS: After ethical approval,patients scheduled for general anaesthesia were randomly allocated to the esketamine group ( n=21; induction with 0.5 mg/kg BW esketamine and 1.5 mg/kg BW methohexitone, ventilation until cord cutting with FiO(2) 1.0) and the sevoflurane-group ( n=21; induction with 1.5-2.0 mg/kg BW methohexitone, ventilation until uterotomia with N(2)O/O(2) [FiO(2) 0,5] and endtidal sevoflurane concentrations about 1.0 vol%). After fetal development, all patients received 2.5 microg/kg BW fentanyl and sevoflurane (about 1.0 vol% endtidal) during ventilation with N(2)O/O(2) (FiO(2) 0.33). As a further control, a group with spinal anaesthesia ( n=22; 2.6-3.0 ml isobaric bupivacaine 0.5 %) was investigated. Maternal circulatory and endocrine plasmatic stress parameters were investigated at five time points and fetal parameters once after development.alpha< or =0.05 was considered significant. RESULTS: Biometric data were comparable in all groups. Systolic arterial pressure was higher in the esketamine group ( p=0.008), whereas the heart rate was lower during spinal anaesthesia ( p<0.001). Plasma noradrenaline decreased in all collectives ( p<0.001) and mean group levels of noradrenaline ( p=0.04) and adrenaline ( p<0.001) were lower during spinal anaesthesia. In all groups, antidiuretic hormone (ADH) remained within the normal range or was slightly increased. Adrenocorticotropic hormone (ACTH) initially increased in all groups and decreased in later time course ( p<0.001). Cortisol increased in all groups ( p<0.001) but group levels were lower during spinal anaesthesia ( p<0.001). In the groups with general anaesthesia, no significant differences in recovery times were obvious, and neither recall nor dream reactions were observed. Postoperative hypoxic incidents (psaO(2) <90%) were comparable between the groups. After spinal anaesthesia, first analgesic demand was later than in the controls ( p=0.001), and the total amount of piritramide was lower ( p=0.02). Nausea and vomiting were more frequent during spinal anaesthesia ( p=0.03). All patients were content with their regimen. Apgar scores, pH-values and adrenaline, noradrenaline and cortisol in plasma were comparable in all groups of children (69 children, 5 gemini). The fetal concentration of esketamine (251 ng/ml) was lower than the corresponding maternal values (493 ng/ml). CONCLUSIONS: When compared with methohexitone, sevoflurane and N(2)O for caesarean section, initial total intravenous anaesthesia with esketamine and methohexitone mediated specific antinociception without negative maternal or fetal effects and not taking invasion kinetics or elimination between uterotomia and cord cutting into consideration. Avoidance of N(2)O allows optimal oxygenation of the fetomaternal unit. Stress protection and hemodynamic responses were well balanced, and intraoperative recall or negative dream reactions were lacking. Superior postoperative pain protection was advantageous after spinal anaesthesia, but in contrast, nausea and vomiting were more frequent in this group. PMID- 12577163 TI - [Postoperative analgesia with tramadol and metamizol. Continual infusion versus patient controlled analgesia]. AB - OBJECTIVE: The study compares the i.v. analgesia of tramadol and dipyrone delivered either as continuous infusion or as patient controlled analgesia (PCA). METHODS: After approval by the local ethics committee and informed consent 203 patients recovering from abdominal surgery were randomly assigned to pain treatment either by PCA or continuous intravenous infusion. Both groups received the same analgesic drug combination of tramadol (20 mg/ml) and dipyrone (200 mg/ml). The PCA bolus was set to 1 ml, the continuous infusion varied between 0-8 ml/h according to individual needs. Demographic and surgery related data were compared between the treatment groups, as well as pain scores, analgesic consumption and response rates over a 48 hour study period.A patient was assessed as non responder when rescue medication was necessary during the study period or the patient rated analgesia as insufficient during the final interview. RESULTS: In total,data of 191 patients (infusion-group: 94, PCA-group: 97) could be analyzed. Tramadol consumption was higher in the infusion group compared to the PCA-group (48 hours: 1009.4+/-494.4 vs. 813.0+/-585.3 mg;p<0.001) with no difference in pain scores. In the infusion group, significantly more interventions were necessary to adjust the infusion rate to individual needs (min max: 1-15 vs. 0-2). The number of non responders amounted to 30 (31.9%) and 18 (18.6%;p=0.03) in the infusion and PCA group, respectively. CONCLUSIONS: PCA with tramadol and dipyrone can be considered an alternative for postoperative pain management and provided a more individualized treatment approach with lower analgesic consumption and more responders compared to a continuous infusion. PMID- 12577164 TI - [Secondary lung surgery following contralateral pneumonectomy. Anaesthesiological considerations]. AB - Although there have been an increasing number of reports on secondary lung surgery following contralateral pneumonectomy in recent years, little information is available about the anaesthesiological management of these patients. We therefore report on a 58-year-old patient who had already undergone a left-sided pneumonectomy and now required a right-sided thoracotomy to remove a recurrent tumour in the right upper lobe. The patient received a total intravenous anaesthesia (propofol, fentanyl) combined with atracurium for muscle relaxation. Following the orotracheal intubation with a Woodbridge tube, the patient was ventilated with the high frequency jet ventilation technique. The jet stream was administered via a catheter placed in the tube. The arterial O(2) saturation during ventilation was always 100%, and arterial CO(2) partial pressure was also normal. No complications occurred during tumour resection from the right upper lobe, and the patient was transferred to the ICU with stable pulmonary and haemodynamic conditions. After 2 h of ventilation, the patient was extubated with a completely expanded lung. The postoperative recovery was uneventful. This case report shows that,presupposing a sufficient pulmonary capacity, secondary lung surgery in previously pneumonectomised patients is feasible without complications given an appropriate anaesthesiological management. PMID- 12577165 TI - [Life-threatening displacement of a tracheostomy tube in a patient with severe angiotensin-converting enzyme inhibitor-induced angioedema]. AB - Angioedema of the lips and the tongue with pharyngeal and laryngeal involvement caused by angiotensin-converting enzyme inhibitors (ACEI) is rare but can cause severe airway compromise and even death due to suffocation. We present the case of a 83-year-old woman with a life-threatening displacement of a tracheostomy tube followed by tension pneumothorax after initial successful treatment of such an airway obstruction by emergency tracheostomy. This case highlights the hazards of tracheostomy tube displacement and is a reminder that where concern of tube dislodgement exists and especially when the possibility of orotracheal intubation is lacking due to upper airway obstruction or difficult airway, permanent epithelized tracheostomy should be performed early to ensure safe and fast tube replacement at any time. PMID- 12577166 TI - [Suicide attempt with high-dose ecstasy]. AB - The case of a 16-year-old female patient is reported, who has taken in 30 tablets of Ecstasy in a suicide attempt. Initially were seen a tachycardia,hyperdynamic circulation, dehydration, leg-myocloni in the awake excited female patient. For prophylaxis of aspiration general anaesthesia and intubation with following gastric lavage were performed. During the intensive care treatment the high blood pressure persisted, although general anaesthesia and anti-hypertensive therapy was continued. After extubation the neurologic status of the patient was normal. Emergency physicians see more and more often Ecstasy-intoxications. Signs of an acute intoxication with Ecstasy are a sympathomimetic reaction with tachycardia,hypertonic blood pressure and cardiac arrhythmia as well as fever, respiratory and renal insufficiency, rhabdomyolysis, cerebral edema and convulsion. A specific antidote does not exist; the symptomatic therapy consists of airway management, hemodynamic stabilisation, primary drug-elimination and, in case of need, application of betablockers and relaxants. PMID- 12577167 TI - [The choice of drugs for caudal anaesthesia in children. An overview]. AB - Caudal anaesthesia is the most frequently used regional technique in paediatric anaesthesia. Caudal in combination with general anaesthesia is usually performed in healthy children (i.e. for herniotomy or hypospadias). Therefore every complication of this method is a catastrophe, even when the incidence of these complications is very low. Some of the documented complications of caudal anaesthesia in children are caused by the local anaesthetic solutions and/or by additives. Thus, the choice of substances for paediatric caudal blocks should minimize the risk associated with the substances used for this indication. Over the last decades the standard was bupivacaine but because of serious cardiovascular and central-nervous toxicity following inadvertent intravascular injection of bupivacaine during caudal puncture, the less toxic ropivacaine should be favoured for this indication. A huge number of clinical studies have proven the clinical effectiveness and safety of ropivacaine also for this indication. In addition, levobupivacaine, the L-enantiomere of bupivacaine, will also be an interesting local anaesthetic in the future for paediatric caudal anaesthesia. By using additives to local anaesthetics better analgesic properties should be obtained. Following an exact review of the literature, only clonidine and S(+)-ketamine are useful additives to local anaesthetics in paediatric caudal anaesthesia. PMID- 12577168 TI - [Ultrasound and local anaesthesia. Part III: ultrasound and neuroaxial local anaesthesia]. AB - The use of ultrasound as a diagnostic tool for the visualisation of the epidural space has effects on the quality and the performance of epidural anesthesia. This work presents an overview of the recent experiences with ultrasound for epidural anesthesia and on the possibilities for ultrasound imaging techniques. The results of visualisation of the epidural space and its limiting structures obtained by various working groups are presented. We review all presently available data on the prediction of the puncture depth. The various working groups found correlations between predicted and effective puncture depth between 0.79 and 0.98 and the precision of the measurement was 57-7.7 mm. Regarding the prediction of the puncture angle there was a poor correlation ranging between 0.07 and 0.31. The precision between the measured and the punctured angles was found to be 10-13.4 degrees. In all available prospective randomised studies on the puncture effects in the lumbar epidural space, the influence of ultrasound showed a significant reduction ( p<0.03) of the puncture attempts,and we found a significant ( p<0.05) reduction in the number of puncture levels. The ultrasound guided puncture allowed an ideal needle trajectory and a more precise application of the catheter. A significant improvement of analgesia quality ( p<0.035) and patient satisfaction ( p<0.006) could be achieved. The metaanalysis of the different studies regarding puncture quality by ultrasound-guided peridural anaesthesia showed a clear advantage for the use of imaging techniques. PMID- 12577169 TI - [Past, present and future of jet ventilation. September 20.-21.2002, Jena]. PMID- 12577170 TI - [Unintentional bupivacaine injection in i.v. regional anesthesia]. PMID- 12577172 TI - [The management of anaesthesia in hepatic resection and cryosurgery of the liver]. PMID- 12577173 TI - Validity of current experimental evidence on laparoscopic surgery for colorectal cancer. PMID- 12577174 TI - Long-term follow-up evaluation for a canine model of gastroesophageal reflux disease. PMID- 12577176 TI - Laparoscopic total mesorectal excision. PMID- 12577179 TI - A comprehensive review of treatments for postmenopausal osteoporosis. AB - The aim of this review is to assess the efficacy of treatments for postmenopausal osteoporosis in women with low bone mass or with an existing vertebral fracture. We searched the literature for studies (randomized, double-masked, placebo controlled and prospective) that reported on drugs registered in Europe or North America. We included 41 reports on 12 agents. To assess the consistency among the studies for each drug, we plotted the percent change in bone mineral density (BMD) for the control group against the percent change in BMD for the treated group for lumbar spine and femoral neck. We used methods of cluster analysis to determine consistency among the studies. For each agent we summarized the relative risk for vertebral fracture (patients with new fracture) and for hip fractures. The duration of the studies ranged from 1 to 4.3 years. The proportion of patients who discontinued treatment ranged from 4% to 80%. Most of the studies reported on change in BMD. Twenty-six studies (10 drugs) provided data on new vertebral fractures and 12 (6 drugs) on hip fractures. Apart from fluoride effects on spine BMD, increases in BMD with bisphosphonates were greater than those seen with the remaining treatments. Generally, for each agent the changes in BMD (relative to placebo) were consistent among the studies. The exceptions were calcitriol and calcitonin for changes in BMD of the spine and of the femoral neck. Alendronate, calcitonin, risedronate and raloxifene caused significant reductions in the risk of vertebral fractures. Alendronate, risedronate or the combination of calcium plus vitamin D had a significant effect on the risk of hip fracture. Most therapies are effective in increasing BMD; some decrease the risk of vertebral fracture. For hip fracture, alendronate and risedronate reduce the risk in women with osteoporosis, and calcium and vitamin D reduce the risk in institutionalized patients. PMID- 12577180 TI - A prospective study of discordance in diagnosis of osteoporosis using spine and proximal femur bone densitometry. AB - The T-score definition of osteoporosis, originally intended for epidemiologic research in this condition, is frequently used in making treatment decisions for individual patients. Discordance in classification depending on the site and type of measurement has been reported in retrospective and cross-sectional studies, but the impact of such discordance on clinical practice is unknown. This is potentially important in view of a recent proposal to confine osteoporosis diagnosis to densitometry at the hip. Having excluded those with degenerative changes in the lumbar spine, we compared the T-score classification of a prospective cohort of patients referred for their first dual-energy X-ray absorptiometry (DXA) scan, analyzing data for men and women in 10-year age groups. Total hip and neck of femur DXA identified significantly fewer osteoporotic patients than spine DXA, and this reduced sensitivity could not be improved by adjusting the T-score threshold without an unacceptable increase in non-osteoporotic cases. The majority of patients undetected by proximal femur DXA were at significantly increased risk of vertebral fracture. DXA at the lumbar spine had only moderate sensitivity and specificity for osteoporosis redefined by total hip densitometry, indicating differential rates of bone loss at the proximal femur and spine. We conclude that, as the most usual indication for bone densitometry is to aid the determination of an individual's fracture risk, both proximal femur and lumbar spine should continue to be assessed. PMID- 12577181 TI - Determinants of incident vertebral fracture in men and women: results from the European Prospective Osteoporosis Study (EPOS). AB - The aim of this analysis was to determine the influence of lifestyle, anthropometric and reproductive factors on the subsequent risk of incident vertebral fracture in men and women aged 50-79 years. Subjects were recruited from population registers from 28 centers across Europe. At baseline, they completed an interviewer-administered questionnaire and had lateral thoraco lumbar spine radiographs performed. Repeat spinal radiographs were performed a mean of 3.8 years later. Incident vertebral fractures were defined morphometrically and also qualitatively by an experienced radiologist. Poisson regression was used to determine the influence of the baseline risk factor variables on the occurrence of incident vertebral fracture. A total of 3173 men (mean age 63.1 years) and 3402 women (mean age 62.2 years) contributed data to the analysis. In total there were 193 incident morphometric and 224 qualitative fractures. In women, an age at menarche 16 years or older was associated with an increased risk of vertebral fracture (RR = 1.80; 95%CI 1.24, 2.63), whilst use of hormonal replacement was protective (RR = 0.58; 95%CI 0.34, 0.99). None of the lifestyle factors studied including smoking, alcohol intake, physical activity or milk consumption showed any consistent associations with incident vertebral fracture. In men and women, increasing body weight and body mass index were associated with a reduced risk of vertebral fracture though, apart from body mass index in men, the confidence intervals embraced unity. For most variables the strengths of the associations observed were similar using the qualitative and morphometric approaches to fracture definition. In conclusion our data suggest that modification of other lifestyle risk factors is unlikely to have a major impact on the population occurrence of vertebral fractures. The important biological mechanisms underlying vertebral fracture risk need to be explored using new investigational strategies. PMID- 12577182 TI - Risk factors associated with peri- and postmenopausal bone loss: does HRT prevent weight loss-related bone loss? AB - In the present study we evaluated the risk factors associated with peri- and postmenopausal bone loss and the effect of hormone replacement therapy (HRT) on weight-loss-related bone loss. The study population, 940 peri- and postmenopausal women, was selected from a random sample (n = 2025) of the OSTPRE study cohort (n = 13 100) in Kuopio, Finland. Bone mineral density (BMD; g/cm(2)) at the lumbar spine and femoral neck, and body weight, were measured at baseline in 1989-91 and at 5-year follow-up in 1994-97 by trained personnel. Five hundred and forty-seven women had never used HRT and 393 women used part-time or continuous HRT during follow-up of 3.8-7.9 years (mean 5.8 years). Similarly, of the 172 weight losers, 97 had never used HRT while 75 used it during follow-up. According to multiple regression analysis on the total study population (n = 940), HRT use, years since menopause and weight increase significantly predicted lower annual bone loss at both the lumbar spine and femoral neck (p < 0.005). Low baseline weight and higher age predicted higher bone loss only at the lumbar spine (p < 0.001) and high grip strength predicted lower bone loss only at the femoral neck (p = 0.021). In a sub-analysis on weight losers, weight loss predicted greater bone loss in HRT non-users (p < 0.05), whereas this was not observed in HRT users. These results remained similar after adjustment for age, weight, height, calcium intake, duration of menopause, baseline BMD and bone-affecting diseases/medication. In conclusion, the transition to menopause, HRT and weight change are the most important determinants of bone loss at both the lumbar spine and femoral neck. Furthermore, HRT seems to be effective in prevention of weight loss related bone loss. PMID- 12577183 TI - Lifestyle factors are associated with osteoporosis in lean women but not in normal and overweight women: a population-based cohort study of 1222 women. AB - The aim of the present population-based cohort study was to evaluate the contribution of lifelong lifestyle factors to calcaneal and distal forearm bone mass in elderly women. We studied 1222 of the 1689 eligible home-dwelling women aged 70-73 years. Lifelong occupational and leisure time physical activity, calcium intake, smoking, alcohol intake and medical history were obtained by a self-completed questionnaire. Main outcome measures were broadband ultrasound attenuation (BUA) of the calcaneus and bone mineral density (BMD) of the radius measured once in 1997-1998. The women with BMI < or = 25.1 kg m(2) had lower BUA (p < 0.0001) and radial BMD values (p < 0.0001) than women with higher BMI. Lifestyle factors associated with BUA in the leanest women were: low physical activity at work (RR 0.4; 95% confidence interval 0.2 to 0.8), low habitual exercise at the ages 30 years, 50 years and currently (RR 1.5; 1.0 to 2.4; RR 1.5; 1.1 to 2.6; RR 1.7; 1.1 to 2.7), poor mobility (RR 1.9; 1.2 to 3.0), coffee intake > or = 5 cups/day (RR 1.7; 1.1 to 2.7), type 2 diabetes (RR 0.3; 0.1 to 0.9) and hypertension (RR 0.5; 0.3 to 0.8). Type 2 diabetes protected lean women from lower distal and ultradistal radial bone density (RR 0.3; 0.1 to 0.8; RR 0.1; 0.1 to 0.5). The selected lifestyle factors were not associated with lowered calcaneal or radial bone density in the higher categories of BMI. In conclusion, risk factors for lower calcaneal and radial bone density appear to be different among lean and normal/obese women. Lifelong recreational physical activity, low physical activity at work, type 2 diabetes and hypertension seem to be associated with increased bone density, while high coffee intake may increase the risk of lower bone density in lean elderly women. These factors are potentially modifiable, and intervention studies targeted at this risk category of women are needed. PMID- 12577184 TI - The association of endogenous hormone concentrations and bone mineral density measures in pre- and perimenopausal women of four ethnic groups: SWAN. AB - We evaluated bone mineral density (BMD), hormone concentrations and menstrual cycle status to test the hypothesis that greater variations in reproductive hormones and menstrual bleeding patterns in mid-aged women might engender an environment permissive for less bone. We studied 2336 women, aged 42-52 years, from the Study of Women's Health Across the Nation (SWAN) who self-identified as African-American (28.2%), Caucasian (49.9%), Japanese (10.5%) or Chinese (11.4%). Outcome measures were lumbar spine, femoral neck and total hip BMD by dual-energy X-ray densitometry (DXA). Explanatory variables were estradiol, testosterone, sex hormone binding globulin (SHBG) and follicle stimulating hormone (FSH) from serum collected in the early follicular phase of the menstrual cycle or menstrual status [premenopausal (menses in the 3 months prior to study entry without change in regularity) or early perimenopause (menstrual bleeding in the 3 months prior to study entry but some change in the regularity of cycles)]. Total testosterone and estradiol concentrations were indexed to SHBG for the Free Androgen Index (FAI) and the Free Estradiol Index (FEI). Serum logFSH concentrations were inversely correlated with BMD (r = -10 for lumbar spine [95% confidence interval (CI): -0.13, -0.06] and r = -0.08 for femoral neck (95% CI: -0.11, -0.05). Lumbar spine BMD values were approximately 0.5% lower for each successive FSH quartile. There were no significant associations of BMD with serum estradiol, total testosterone, FEI or FAI, respectively, after adjusting for covariates. BMD tended to be lower (p values = 0.009 to 0.06, depending upon the skeletal site) in women classified as perimenopausal versus premenopausal, after adjusting for covariates. Serum FSH but not serum estradiol, testosterone or SHBG were significantly associated with BMD in a multiethnic population of women classified as pre- versus perimenopausal, supporting the hypothesis that alterations in hormone environment are associated with BMD differences prior to the final menstrual period. PMID- 12577185 TI - Hospital care of osteoporosis-related vertebral fractures. AB - Resource implications of hospitalization for osteoporosis-related vertebral fracture are sparsely documented. This study utilized data abstracted from a national sample of hospitalized patients to identify characteristics of patients who are hospitalized with vertebral fracture and their patterns of resource utilization. These were compared with patterns observed for hip fracture hospitalizations. Data from the Nationwide Inpatient Sample (NIS) for 1997 were used to identify men and women age 45 years and above who had a primary diagnosis of vertebral fracture. After patients whose fractures might have been due to metastatic cancer or severe trauma were excluded, 68,901 individuals hospitalized for vertebral fracture were identified. Seventy-seven percent of these were women, most were white, 75 years and older, and had multiple comorbid diagnoses. Total charges averaged 8000-10,000 US dollars per hospitalization and were higher in men. Mean length of stay was just under 6 days and more than 50% of discharged patients required some form of continuing care. Hospitalizations for vertebral fracture occurred at only one-fourth the rate of those for hip fracture, and created only half the hospital charges per admission. Vertebral fracture accounted for over 400,000 total hospital days and generated charges in excess of 500 million US dollars. This resource impact is considerably higher than has been described in prior studies. PMID- 12577186 TI - Prevalent vertebral deformities predict increased mortality and increased fracture rate in both men and women: a 10-year population-based study of 598 individuals from the Swedish cohort in the European Vertebral Osteoporosis Study. AB - The aim of this study was to evaluate whether a prevalent vertebral deformity predicts mortality and fractures in both men and women. In the city of Malmo, 598 individuals (298 men, 300 women; age 50-80 years) were selected from the city's population and were included in the Swedish part of the European Vertebral Osteoporosis Study (EVOS). At baseline the participants answered a questionnaire and lateral spine radiographs were performed. The prevalence of subjects with vertebral deformity was assessed using a morphometric method. The mortality during a 10-year follow-up period was determined through the register of the National Swedish Board of Health and Welfare. Eighty-five men and 43 women died during the study period. The subsequent fracture incidence during the follow-up period was ascertained by postal questionnaires, telephone interviews and by a survey of the archives of the Department of Radiology in the city hospital. Thirty-seven men and 69 women sustained a fracture during the study period. Data are presented as hazard ratios (HR) with 95% confidence interval (95% CI) within brackets. Prevalent vertebral deformity, defined as a reduction by more than 3 standard deviations (SD) in vertebral height ratio, predicted mortality during the forthcoming decade in both men [age-adjusted HR 2.4 (95% CI 1.6-3.9)] and women [age-adjusted HR 2.3 (95% CI 1.3-4.3)]. In men there was an increased mortality due to cardiovascular and pulmonary diseases and in women due to cancer. Prevalent vertebral deformity predicted an increased risk of any fracture during the forthcoming decade in both men [age-adjusted HR 2.7 (95% CI 1.4-5.3)] and women [age-adjusted HR 1.8 (95% CI 1.1-2.9)]. Prevalent vertebral deformity predicted an increased risk of any subsequent fragility fracture in women [age adjusted HR 2.0 (95% CI 1.1-3.5)]; however, in men the increased risk was nonsignificant [age-adjusted HR 1.9 (95% CI 0.7-5.1)]. In summary, a prevalent vertebral deformity can predict both increased mortality and increased fracture incidence during the following decade in both men and women. We conclude that prevalent vertebral deformity could be used as a risk factor in both genders for mortality and future fracture. PMID- 12577187 TI - Disability after clinical fracture in postmenopausal women with low bone density: the fracture intervention trial (FIT). AB - Relatively little is known about outcomes following clinical osteoporotic fractures at nonhip, nonvertebral skeletal sites. To address this issue, we prospectively assessed post-fracture disability at multiple skeletal sites in a population of 909 older (aged 55-81 years), community-dwelling women with low femoral neck bone mineral density who had experienced a fracture while enrolled in the Fracture Intervention Trial (FIT). FIT is a randomized, double-masked, placebo-controlled trial that was designed to determine the effect of alendronate on fracture incidence, and the current study was conducted as a secondary analysis of FIT data. Following incident clinical fractures, FIT participants were followed prospectively for assessment of site-specific, fracture-related disability. Measures of disability were self-reported days hospitalized or confined to bed because of the fracture ('bed days') and days of reduced usual activities because of the fracture ('limited activity days'). Of fracture types evaluated, those of the hip resulted in the highest percentage of subjects with any bed days or limited activity days after fracture (94% with any bed days and 100% with any limited activity days), though the mean number of bed days and limited activity days appeared highest after lumbar vertebral fractures (25.8 mean bed days and 158.5 mean limited activity days). Substantial disability also was reported after fractures of thoracic vertebrae, humerus, distal forearm, ankle and foot. Within fracture types, post-fracture disability was highly variable, ranging from none to more than 6 months. PMID- 12577188 TI - Daily treatment with parathyroid hormone is associated with an increase in vertebral cross-sectional area in postmenopausal women with glucocorticoid induced osteoporosis. AB - Daily treatment with hPTH (1-34) is associated with a significant increase in bone formation which results in large gains in lumbar spine bone mass. However, bone formation is known to occur on trabecular, endocortical and periosteal surfaces. The purpose of this study was to determine whether daily treatment with hPTH (1-34) for 1 year was associated with a change in vertebral cross-sectional area, or vertebral size, as measured by serial quantitative computed tomography scans. Fifty-one postmenopausal women treated chronically with both glucocorticoids and hormone replacement therapy (HRT) were randomized to either daily hPTH (1-34) for 1 year and HRT or to a control group treated with only HRT. Measurements of bone density of the spine were obtained every 6 months by dual energy X-ray absorptiometry (DXA) and annually by QCT of the L1 and L2 vertebrae. Vertebral cross-sectional area (VCSA) was obtained from the QCT scans. In addition, we estimated the vertebral compressive strength (VSFOM, g(2)/cm(4) = trabecular BMD(2) x VCSA). After 1 year of hPTH (1-34) treatment, VCSA increased 4.8% (p < 0.001), and 1 year after treatment was discontinued VCSA was still 2.6% higher than the baseline value (p < 0.05). The control group had no change in VCSA. In addition, estimated vertebral compressive strength increased more than 200% over baseline levels in the hPTH (1-34) treatment group and no change was observed in the control group. In summary, daily treatment with hPTH (1-34) for 1 year increased vertebral size as measured by VCSA and this increase was maintained after hPTH (1-34) was discontinued. Since vertebral fracture risk is related to both bone size and bone mass, we cautiously speculate that the increase in vertebral size associated with hPTH (1-34) treatment is at least partially responsible for increased vertebral bone strength and reduction of fracture risk associated with this therapy in postmenopausal osteoporosis. PMID- 12577190 TI - The effects of alendronate in postmenopausal women with osteoporosis. PMID- 12577189 TI - Better late than never? Experience with intravenous pamidronate treatment in patients with low bone mass or fractures following cardiac or liver transplantation. AB - Organ transplantation is associated with a high turnover of bone metabolism, and an increased loss of bone mass and incidence of osteoporotic fractures. Established therapies for osteoporosis after organ transplantation are still lacking, however. We report on an intravenous bisphosphonate therapy initiated in transplant patients because of a high rate of bone loss or incident osteoporotic fractures. Twenty-one patients after liver transplantation and 13 patients after heart transplantation received 30 mg pamidronate intravenously every 3 months, combined with 1000 mg calcium and 1000 IU vitamin D per day. The median time interval between transplantation and start of pamidronate treatment was 1.9 years in cardiac patients and 2.3 years in liver patients. Lumbar spine bone mineral density (LS BMD) and femoral neck BMD (FN BMD) were measured before and every 6 months after pamidronate therapy was initiated. Spinal radiographs were performed annually. Biochemical markers of bone metabolism were determined every 3 months, immediately before pamidronate administration. From a previous observational study, 58 patients treated only with calcium and vitamin D were matched for age, sex, pretransplantation LS BMD and time interval between transplantation and the first pamidronate treatment. In the pamidronate-treated patients, the mean increase in LS BMD adjusted for baseline values amounted to 0.080 +/- 0.038 g/cm(2) (8.6 +/- 4.0 %) after 1 year and 0.091 +/- 0.058 g/cm(2) (10.4 +/- 6.1%) after 2 years compared with 0.001 +/- 0.037 g/cm(2) (0.26 +/- 4.0%) after 1 year and 0.015 +/- 0.057 g/cm(2) (1.8 +/- 6.0%) after 2 years in the historical control group (absolute LS BMD changes pamidronate group vs historical group p < 0.0001 after 1 and 2 years). The changes of FN BMD were 0.024 +/- 0.043 g/cm(2) (3.2 +/- 6.1%) after 1 year and 0.046 +/- 0.052 g/cm(2) (7.0 +/- 6.1%) after 2 years in the pamidronate group compared with -0.012 +/- 0.043 g/cm(2) (-1.6 +/- 6.1%) after 1 year and -0.013 +/- 0.052 g/cm(2) (-1.1 +/- 6.1%) after 2 years in the historical control group (absolute FN BMD changes pamidronate group vs historical group p = 0.003 after 1 year and p = 0.001 after 2 years). From a total of 287 application cycles of pamidronate treatment, no severe side effects were observed and non-severe side effects were seen in only 39 cycles (13.6%). We conclude that cyclic intravenous pamidronate treatment is beneficial to patients with low bone mass or osteoporotic fractures following transplant, even when not immediately initiated. PMID- 12577191 TI - Sequencing to NRTI plus NNRTI-only combinations after virological failure of protease inhibitor-based combination HIV-1 therapy. AB - PURPOSE: To examine the success of sequencing to nucleoside reverse transcriptase inhibitor (NRTI) plus nonnucleoside reverse transcriptase inhibitor (NNRTI)-only combinations after virological failure of protease inhibitor (PI)-based combinations in NNRTI-naive individuals. METHOD: This was an observational cohort study. RESULTS: 171 patients were identified. The group was highly antiretroviral therapy-experienced with median cumulative NRTI and PI durations prior to change of 53 months and 21 months, respectively. The median CD4 count and viral load (VL) prior to change were 228 cells/mm(3) and 24500 copies/mL. Overall, 37.4% had a VL below the limit of detection (50 copies/mL) at 12 months (intention-to-treat analysis). Markers associated with success at 12 months were efavirenz (EFV) use, the use of three NRTI+NNRTI combinations, and abacavir (ABC) use. In multivariate analysis, use of EFV remained highly significant (relative hazard of virological success of nevirapine [NVP] vs. EFV = 0.39; p =.003), while ABC use became nonsignificant. A benefit from the use of three NRTI + NNRTI regimens was observed. CONCLUSION: NRTI plus NNRTI-only combinations performed poorly, however the superior outcomes of patients treated with EFV are consistent with findings in cohort studies. The suggestion of benefit in patients receiving three NRTIs in addition to either NVP or EFV deserves further study in randomized trials. PMID- 12577192 TI - First-line therapy and mitochondrial damage: different nucleosides, different findings. AB - BACKGROUND: Antiretroviral therapy has been associated with the development of morphologic body-shape changes and metabolic abnormalities, including dislipemia, insulin resistance, and hyperlactatemia. Mitochondrial damage secondary to the use of nucleoside analogue reverse transcriptase inhibitors (NRTIs) has been related to some of these complications, although the role of different NRTIs in their development is not well established. OBJECTIVES: To assess the incidence of hyperlactatemia and lipodystrophy body-shape changes in drug-naive HIV-infected patients who began highly active antiretroviral therapy (HAART) based on a backbone of two different NRTI combinations. METHOD: Prospective, longitudinal, observational study of all consecutive drug-naive HIV-infected individuals who started HAART with zidovudine (AZT) plus lamivudine (3TC) or didanosine (ddI) plus stavudine (d4T) between June 2000 and June 2001 at one single institution. Serum lactate levels and lipodystrophy body-shape changes were monitored periodically during 12 months. RESULTS: At 1 year, mean lactate values remained <2 mmol/L in all 26 patients who received AZT+3TC, but they significantly increased (mean, 2.6 mmol/L) in 50 patients treated with ddI+d4T. The percentage of patients with hyperlactatemia (lactate >or=2 mmol/L) steadily increased in those on ddI+d4T (from 30% at 3 months to 71% at 12 months), whereas it remained below 10% in patients treated with AZT+3TC. Two patients on ddI+d4T developed lactic acidosis. Mean serum lactate dehydrogenase (LDH), gamma glutamyltransferase (GGT), and amylase significantly increased in patients treated with ddI+d4T, whereas they remained unaltered in patients under AZT+3TC. Significant correlations were found between lactate and LDH, alkaline phosphatase (AP), and GGT. In the multivariate analysis, treatment with ddI+d4T, LDH, and AP was significantly associated with lactate levels. At 12 months, subcutaneous lipoatrophy was significantly more frequent in patients treated with ddI+d4T than in those on AZT+3TC (35% vs. 8%; p =.01). CONCLUSION: In drug-naive HIV-infected patients who start antiretroviral therapy, ddI+d4T-based combinations produce a greater increase in serum lactate and lipoatrophy than therapies based on AZT+3TC within the first year of therapy. An increase in LDH, amylase, GGT, and AP levels may signal an increase in lactate, which may be harmful. PMID- 12577193 TI - Overcoming resistance: virologic response to a salvage regimen with the combination of ritonavir plus indinavir. AB - PURPOSE: To explore the possibility of overcoming resistance to protease inhibitors (PIs) and to determine the resistance cutoff values that continue to predict treatment failure with a dual PI regimen. METHOD: We performed a prospective study of 53 patients who had failed in several PIs and who were included in a ritonavir (RTV) plus indinavir (IDV) salvage regimen. Median HIV RNA level decrease was evaluated according to resistance assays and indinavir trough levels. RESULTS: Eighty-seven percent of patients had previously failed on an IDV-containing regimen. Overall, median HIV RNA decrease was -1.25 log(10) copies/mL after 3 months on therapy. A significant blunted virologic response was observed only in isolates with more than 12 substitutions including the V82A ( 0.75 vs. -1.3 log(10) copies/mL; p =.04), or in isolates with more than 30 fold increase in the IC(50) (-0.43 vs. -1.2 log(10) copies/mL). Higher drug levels were observed in patients with resistant isolates who achieved an HIV RNA decrease greater than 1 log (1742 vs. 1100 ng/mL). CONCLUSION: Our preliminary data suggest the possibility of overcoming resistance with the combination of RTV plus IDV. They also suggest the need for establishing new resistance cutoff values when using PIs in combination. PMID- 12577194 TI - Lipid changes in patients initiating efavirenz- and indinavir-based antiretroviral regimens. AB - PURPOSE: To evaluate nonfasting lipid levels in a large cohort of patients on three HAART regimens: efavirenz + zidovudine + lamivudine (EFV+ZDV+3TC), efavirenz + indinavir (EFV+IDV), and indinavir + zidovudine + lamivudine (IDV+ZDV+3TC). METHOD: Nonfasting lipid levels were analyzed from a large randomized multicenter treatment trial for HIV-infected patients initiating HAART. Treatment evaluations were carried out at prescribed intervals, and data were recorded and analyzed. Assessment was limited to high-density lipoprotein (HDL) and total cholesterol. RESULTS: The results demonstrate an increase in the total cholesterol, ranging from 23 to 57 mg/dL, in the three combinations of HAART therapy. The increase was most significant in the EFV+IDV arm where the effects appear to be additive. HDL cholesterol also increased in all three arms, but the greatest increase was in the two groups containing EFV. In all three arms, the HDL cholesterol increased significantly in women while increases in men were seen only in the EFV-containing arms. Men taking either IDV-containing regimen had a greater increase in total cholesterol, and therefore the total/HDL cholesterol ratio rose significantly. CONCLUSION: EFV and IDV independently elevate lipid levels. Alterations in the lipid levels may lead to increased cardiovascular risk in men, possibly mitigated by elevations in HDL cholesterol. In addition, changes in HDL cholesterol were significantly different between men and women. PMID- 12577195 TI - Comparison of metabolic abnormalities and clinical lipodystrophy 48 weeks after switching from HAART to Trizivir versus continued HAART: the Trizal study. AB - PURPOSE: To analyze the evolution of clinical lipodystrophy (LD) and metabolic abnormalities in patients continuing to receive HAART versus patients switched to Trizivir (zidovudine, lamivudine, abacavir) after 48 weeks. METHOD: Patients treated with HAART >6 months with plasma HIV-1 RNA viral load (VL) <400 copies/mL and <50 copies/mL at screening were randomly assigned to continue HAART (103 patients) or to receive Trizivir (106 patients). Clinical LD was evaluated using a standardized patient questionnaire only at baseline, weeks 4 and 8, and then every 8 weeks until Week 48. Laboratory evaluation was performed every 4 weeks. RESULTS: The proportion of patients exhibiting >or=1 LD symptom at baseline was 40% in the Trizivir arm and 50% in HAART arm (difference not significant). After 48 weeks, the prevalence was 28% and 42% respectively (p =.03), and the median number of LD symptoms per patient was 2 in the Trizivir arm and 4 in the continued HAART arm (p =.016). Median decreases in cholesterol levels over the 48 week study period were greater in the Trizivir arm than in the continued HAART arm (-0.80 vs. -0.44 mmol/L; p lt.001). Median triglyceride levels decreased in the Trizivir arm but increased in the continued HAART arm (-0.17 and +0.01 mmol/L; p =.006). Suppression of VL was maintained in most patients with no differences between the two arms. CONCLUSION: A switch from "standard" HAART to Trizivir was associated with an improvement in clinical LD and blood lipid abnormalities after 48 weeks. PMID- 12577196 TI - Centralized assessment of dual-energy X-ray absorptiometry (DEXA) in multicenter studies of HIV-associated lipodystrophy. AB - BACKGROUND: The accurate measurement of total body and subcutaneous fat is essential if therapeutic interventions, aimed at preventing or reversing lipodystrophy syndrome, are to be adequately assessed. The aim of this study was to investigate the variability of dual-energy X-ray absorptiometry (DEXA) scans analysis performed at local sites compared to central analysis in a multicenter clinical trial. METHOD: The PIILR study was a multicenter randomized clinical trial in which 80 HIV-infected patients with physician-documented lipodystrophy had serial measurements of body composition performed with Lunar DEXA scans. Scans were analyzed at local sites and then were reanalyzed centrally. RESULTS: DEXA scans from 73 patients who completed 24 weeks study were compared. Greater variation in the locally analyzed results than in the centrally reanalyzed data was noted, with arm, leg, and combined limb fat being most divergent between the local and centralized assessments (ratio of local to central standard deviation was 1.28, 1.31, and 1.35, respectively). The magnitude of this variance was enough to alter statistically relevant differences between study populations. CONCLUSION: Quality assurance is an important issue in the use of DEXA scans to determine body fat composition in multicenter research studies. A central quality assurance site should be incorporated to reduce variability in results. PMID- 12577197 TI - Selected topics from the Sixth International Congress on Drug Therapy in HIV infection; November 17-21, 2002; Glasgow, United Kingdom. PMID- 12577198 TI - National Institutes of Health Consensus Development Conference statement Management of Hepatitis C: 2002 June 10-12, 2002. PMID- 12577199 TI - Hepatitis B virus infection despite receiving lamivudine in one HIV-infected person. PMID- 12577200 TI - Tissue-specific reduction in splicing efficiency of IKBKAP due to the major mutation associated with familial dysautonomia. AB - We recently identified a mutation in the I-kappa B kinase associated protein (IKBKAP) gene as the major cause of familial dysautonomia (FD), a recessive sensory and autonomic neuropathy. This alteration, located at base pair 6 of the intron 20 donor splice site, is present on >99.5% of FD chromosomes and results in tissue-specific skipping of exon 20. A second FD mutation, a missense change in exon 19 (R696P), was seen in only four patients heterozygous for the major mutation. Here, we have further characterized the consequences of the major mutation by examining the ratio of wild-type to mutant (WT:MU) IKBKAP transcript in EBV-transformed lymphoblast lines, primary fibroblasts, freshly collected blood samples, and postmortem tissues from patients with FD. We consistently found that WT IKBKAP transcripts were present, albeit to varying extents, in all cell lines, blood, and postmortem FD tissues. Further, a corresponding decrease in the level of WT protein is seen in FD cell lines and tissues. The WT:MU ratio in cultured lymphoblasts varied with growth phase but not with serum concentration or inclusion of antibiotics. Using both densitometry and real-time quantitative polymerase chain reaction, we found that relative WT:MU IKBKAP RNA levels were highest in cultured patient lymphoblasts and lowest in postmortem central and peripheral nervous tissues. These observations suggest that the relative inefficiency of WT IKBKAP mRNA production from the mutant alleles in the nervous system underlies the selective degeneration of sensory and autonomic neurons in FD.Therefore, exploration of methods to increase the WT:MU IKBKAP transcript ratio in the nervous system offers a promising approach for developing an effective therapy for patients with FD. PMID- 12577204 TI - [Towards normality (II)]. PMID- 12577203 TI - [Towards normality]. PMID- 12577201 TI - Extensive linkage disequilibrium, a common 16.7-kilobase deletion, and evidence of balancing selection in the human protocadherin alpha cluster. AB - Regions of extensive linkage disequilibrium (LD) appear to be a common feature of the human genome. However, the mechanisms that maintain these regions are unknown. In an effort to understand whether gene density contributes to LD, we determined the degree of promoter sequence variation in a large tandem-arrayed gene family, the human protocadherin alpha cluster, on chromosome 5. These genes are expressed at synaptic junctions in the developing brain and the adult brain and may be involved in the determination of synaptic complexity. We sequenced the promoters of all 13 alpha protocadherin genes in 96 European Americans and identified polymorphisms in the promoters alpha 1, alpha 3, alpha 4, alpha 5, alpha 7, alpha 9, alpha 11, and alpha 13. In these promoters, 11 common SNPs are in extensive LD, forming two 48-kb haplotypes of equal frequency, in this population, that extend from the alpha1 through alpha 7 genes. We sequenced these promoters in East Asians and African Americans, and we estimated haplotype frequencies and calculated LD statistics for all three populations. Our results indicate that, although extensive LD is an ancient feature of the alpha cluster, it has eroded over time. SNPs 3' of alpha 7 are involved in ancestral recombination events in all populations, and overall alpha-cluster LD is reduced in African Americans. We obtained significant positive values for Tajima's D test for all alpha promoter SNPs in Europeans (D=3.03) and East Asians (D=2.64), indicating an excess of intermediate-frequency variants, which is a signature of balancing selection. We also discovered a 16.7-kb deletion that truncates the alpha 8 gene and completely removes the alpha 9 and alpha 10 genes. This deletion appears in unaffected individuals from multiple populations, suggesting that a reduction in protocadherin gene number is not obviously deleterious. PMID- 12577202 TI - A novel NOD2/CARD15 haplotype conferring risk for Crohn disease in Ashkenazi Jews. AB - Crohn disease (CD) exhibits a 2-4-fold increased frequency in Jews as compared with other ethnic/racial groups. Three coding variants of the NOD2/CARD15 have been reported as independent disease-predisposing mutations (DPMs), but these were found in only 30%-40% of patients with CD and could not account for all the linkage between CD and the IBD1 locus. The aim of the present study was to explore whether additional DPMs at the IBD1 locus exist in the high-risk Jewish group. Sixty-four Ashkenazi Jewish and 147 non-Jewish white families were studied. Six microsatellite markers spanning IBD1 were genotyped for linkage analysis in subgroups stratified on NOD2/CARD15 DPM status. SNPs in NOD2/CARD15 (R702W, G908R, 1007fs, and S268P) were then genotyped in family and independent case-control samples. On the basis of initial results, sequencing was done on NOD2/CARD15-translated regions in 12 Jewish individuals. Subsequently, a new NOD2/CARD15 variant was genotyped and analyzed. After excluding the influence of the three DPMs, significant linkage of IBD1 to CD in Jews remained with two peaks at D16S403 (mean allele sharing [MAS] = 0.70] and D16S411 (MAS = 0.59). Further, we observed an increased frequency of a haplotype carrying only the 268S variant in Jewish patients (OR = 3.13, P=.0023) but not in non-Jews, suggesting the existence of a Jewish-specific additional disease-predisposing factor on this haplotype. Sequencing of this haplotype revealed a new variant (IVS8+158; JW1). The 268S-JW1 combination exhibited a further increased risk (OR = 5.75, P=.0005) and the highest population-attributable risk (15.1%) for CD among reported DPMs in Jews. In Ashkenazi Jews, unrecognized population-specific predisposing factor(s) exist on the 268S-JW1 haplotype at the IBD1 locus. This factor may contribute to the higher risk for CD in Ashkenazi Jews as compared with non-Jews. PMID- 12577205 TI - [Neuropsychological disorders in amyotrophic lateral sclerosis. Don't they exist or do they just go undetected?]. AB - AIMS: The aims of this paper is to demonstrate the existence of neuropsychological disorders in amyotrophic lateral sclerosis (ALS) and to perform an in depth study of the cognitive functioning of the prefrontal lobes. PATIENTS AND METHODS: A neuropsychological study of 14 patients with ALS was conducted using an extensive battery of tests and were compared with a group of 14 healthy controls. Both groups were homogeneous as regards age, sex, education and manual dominance. In this clinical and research study, as well as the neuropsychological variables (subtest of the Barcelona PIEN Test neuropsychological battery), we also took the evolution of the disease, the age and neurological clinical features of the patients suffering from ALS into account. RESULTS: We found neuropsychological disorders in the ALS patients that were centred, either directly or indirectly, on functions of the prefrontal lobe, and in particular of the dorsolateral and premotor cortices, which had already been observed in other research work. No memory disorders were found, something which is usually mentioned in studies about neuropsychological disorders in this type of patients. CONCLUSIONS: Apart from the primary motor zones affected in ALS, there appears to be a degenerative process in most of the frontal lobe, and there is a need for longitudinal studies of the possible progressive disorders of the frontal lobe in these patients. This is difficult, since these patients end up with serious neurological alterations which prevent a correct neuropsychological exploration from being carried out cognitively, and hence no conclusions can be drawn either PMID- 12577207 TI - [Peripheral nerve conduction studies in patients with multiple organ failure]. AB - INTRODUCTION: The polyneuropathy of critical diseases that often accompanies multiple organ failure (MOF) is one of the various causes that leads to the death of these patients. AIMS: To evaluate the electrophysiological behaviour of peripheral nerves in patients with MOF. Subjects and methods. A prospective cross sectional study was performed in 18 patients with MOF (the study group), 50 healthy subjects and 20 suffering from Guillain Barr Stroll Landry syndrome (GBS) (the control groups). All of them were submitted to sensory and motor peripheral nerve conduction studies. RESULTS: The aetiology of this syndrome was varied: 17 patients required mechanical ventilation and 15 had difficulty in weaning. The main electrophysiological alterations in the study group consisted in a drop in amplitude with prolonged latency and duration, and slowed conduction velocities for the sensory potentials of the median and posterior tibial nerves, while the lowered amplitude in motor nerve conduction was detected to a lesser degree and intensity. On comparing them with those obtained in GBS, statistically significant differences were found. The greatest electrophysiological alterations were detected in those who had difficulty in weaning those who presented an unfavourable evolution. CONCLUSIONS: Subjects with MOF were found to have a predominantly sensory peripheral axono myelinopathy which, on monitoring it electrophysiologically, allows us to establish a prognosis regarding severity, evolution and gradually cutting the dependence on mechanical ventilation PMID- 12577206 TI - [Quality control in carotid surgery. Results from the carotid surgery records in Central region of Spain (1999-2000)]. AB - OBJECTIVE: Evaluation of technique and results of carotid endarterectomy (CEA) in the central region in Spain and possible differences among the hospitals included. Study of factors predictors of complications. PATIENTS AND METHODS: All CEAs performed in 9 hospitals in the central region in Spain (1999 2000). Registry design by a specific commission and contents selected by consensus. Voluntary participation. Ambispective study. Uni and multivariate statistical analysis (logistic regression). RESULTS: 576 procedures registered. Global mortality: 1.9%. 15 (2.7%) patients died or were severely handicapped. The selection of patients adjusted to expert s recommendations in 98%. Significant differences in technique and results were found among the hospital centres (>0,25). In the multivariate analysis, the risk factors associated with complications were: presence of preoperative symptoms (p= 0.006) small number of annual CEAs (p= 0.01), contralateral occlusion (RR= 4.7, p= 0.02) and use of shunt (RR= 6.1, p= 0.01). CONCLUSIONS: Globally, both the selection of patients and the results obtained adjust to the accepted standards for this type of surgery. The patient s risk profile has had a greater influence on the results than the technique used. The mortality rate has been influenced by the low volume hospitals PMID- 12577208 TI - [Intergenerational study of the mutation that causes Myotonic Dystrophy Type 1 in Costa Rica]. AB - INTRODUCTION: Myotonic dystrophy type 1 is a neuromuscular, degenerative and progressive disease, with an autosomal dominant pattern of inheritance, variable expressivity and incomplete penetrance. The genetic defect is an unstable mutation due to the expansion of the triplet CTG in the 3 unstranslated region at the DMPK gene on chromosome 19q13.3. OBJECTIVE: The main objective was to study the intergenerational behavior of the DM1 mutation in order to evaluate the importance of this disease as a neurological problem that could be manageable by genetic counseling. PATIENTS AND METHODS: The study involved 84 patients with clinical diagnosis of DM1 and their relatives, which were confirmed through molecular diagnosis using Southern blot and PCR. RESULTS: Data analysis reveals the size of the mutation presents a positive correlation with the severity of the symptoms and a negative correlation with the age of onset. Transmission of the DM1 mutation is sex and size dependent among the Costa Rican patients. There is an important increment in the size of the mutation between generations and there are no differences in mutation size respect to the transmitting sex. CONCLUSION: The worldwide intergenerational behavior of the DM1 mutation is similar in Costa Rica PMID- 12577209 TI - [Corrected cerebrospinal fluid manometry by measurement of the density and its relation with its biochemical composition]. AB - INTRODUCTION AND AIMS: We question the suitability of the usual methods for taking the pressure of cerebrospinal fluid (CSF). We suspect that the composition of CSF and, in consequence, its density have an important effect on its manometric value. MATERIAL AND METHODS: We obtained 322 CSF samples, with known cytological and biochemical composition, density and manometric values. We corrected the manometric measurements and, for each case, we calculated the exact pressure exerted by the column of CSF from its density, which was already known. The working variable was set as the manometric error (ME) that resulted from: Abs (calculated pressure of the column of CSF from its density manometry) (absolute value of the difference). RESULTS: It was found that protein concentration levels in CSF had an important effect on the ME of the CSF, especially in values above 150 mg/cm3, and this result is extrapolated from our ad hoc mathematical model: corrected manometry (mmH2O) = manometry (mmH2O) + [cerebrospinal fluid protein (mg/cm3) / 50] 2. CONCLUSION: This model proved to be a suitable predictor of ME in our sample, with a= 0.005. PMID- 12577210 TI - [Disturbances of neuroblast migration and amniocentesis. An under diagnosed problem?]. AB - INTRODUCTION: Performing amniocentesis between the fourth and sixth months of gestation is a widespread practice. However, it can entail serious consequences, apart from the loss of the foetus, such as injuries to the developing central nervous system. CASE REPORTS: Over the past few years we have dealt with four patients who were carriers of a focal disturbance of neuroblast migration, which could have its origins in amniocentesis. Two patients displayed mental retardation and difficult to treat epilepsy, while the other two only presented hemiparesis. In two cases the right hemisphere of the brain was affected. One of them also presented extracranial brain tissue, which was possibly extruded during the accidental puncture of the skull. CONCLUSIONS: Despite its being rarely mentioned in the literature, a direct or indirect attack on the brain during diagnostic amniocentesis is a possibility that must be borne in mind when considering the use of this technique PMID- 12577211 TI - [Neonatal convulsions caused by incontinentia pigmenti with left opercular dysgenesia]. AB - AIMS: In this paper we review the main publications on incontinentia pigmenti (IP) and the current knowledge of the etiopathogenesis of the disease and of the convulsions in the neonatal period, by considering a clear case of neonatal IP, with skin, eye, brain and bone lesions. CASE REPORT: Our patient, a female, started with clonic seizures in the right half of the body at the age of three days. Method. IP, or Bloch Sulzberger syndrome, is a genetic multisystemic neuroectodermic disorder. It is a disease of low incidence (1% of all neuroectodermic disorders) which is transmitted by means of a pattern of dominant inheritance linked to X, and is lethal in males, except in rare cases of somatic mosaicism and Klinefelter. In the family forms the gene is located in the p11 (IP 1) and q28 (IP 2) regions of the X chromosome. It has recently been discovered that the cause lies in a mutation of a gene called NEMO (IKK gamma). Together with Bourneville s tuberous sclerosis it is the only neurocutaneous syndrome that can begin with neonatal convulsions. The convulsions start on the second or third day of life and are often limited to a single side of the body, although it can also appear as encephalitis. The origin of the convulsions has been linked with recurring encephalomyelitis, or with an alteration of the neuronal migration. CONCLUSIONS: The cause of the early convulsions in our patient, which we put down to a left perisylvian focal dysgenesia (unilateral opercular syndrome) observed in the computerised axial tomography (CAT scan), has not been reported up to the present associated with IP. PMID- 12577212 TI - [Island of Reil and pharmacoresistance in epilepsy]. AB - INTRODUCTION: Over the last few years surgery has been developing, with promising results, methods to treat an important number of cases of partial epilepsy that are related, to different extents, with the lobe of the insula and display pharmacoresistance. Better knowledge of the anatomofunctional particularities of this region of the cortex, new neuroimaging and neurophysiological techniques, together with the use of the surgical microscope, stereotactic support and neuronavigation, have had a strong influence on the development of this type of surgery. CASE REPORT: In this paper we report three cases of patients with epilepsy and who were diagnosed as suffering from structural lesions of the insular region: two cavernomas and an oligodendroglioma. The preoperative study was carried out using magnetic resonance imaging, angiography and video EEG. The surgical procedure consisted in a transsylvian approach with electrocorticography, which provided us with the clinical results we have studied. CONCLUSIONS: The correct approach to the diagnosis of the epileptogenic insular lesions must include a complete neurophysiological study and preoperative planning with angiography. This allows their surgical resection through the transsylvian approach and enables us to obtain, with a very low degree of morbidity, clearly satisfactory results as regards the decrease in the number of seizures in these patients PMID- 12577213 TI - [Stenting without prior dilatation in intracranial stenosis of the internal carotid artery: a case report]. AB - INTRODUCTION: Recent reports have described the application of coronary stents for the treatment of intracranial stenosis of the internal carotid artery (ICA), above all in patients who do not respond to medical treatment and display advancing neurological symptoms. Stenting in intracranial vascular lesions of the carotid territory has been used almost exclusively in the treatment of the complications due to transluminal angioplasties with balloons. In selected cases and without prior dilatation of the stenosis it would be possible to place a stent. CASE REPORT: We describe a case of intracranial carotid stenting, without previous dilatation, in a 57 year old male patient with multiple arteriosclerotic risk factors. The patient presented neurological symptoms with no response to medical treatment, caused by a stenosis that affected over 70% of the cavernous segment of the right ICA, which was shown up by the arteriographic study. A favourable anatomy and the material used allowed the stent to pass through the stenosis without the need for dilatation. No immediate complications were observed and the carotid artery was seen to be of a normal calibre for stenosis. The brain angiogram also proved to be normal. The patient continues with the medical treatment and has remained neurologically stable throughout the 9 month clinical follow up. CONCLUSION: In certain selected cases, placing an intracranial carotid stent, without previous balloon dilatation, can give good results and prevent the complications that can accompany angioplasty (such as dissection, rupture or embolism) PMID- 12577214 TI - [The neural substrate of biological rhythms]. AB - INTRODUCTION AND METHOD: The various bodily functions vary rhythmically along the 24 hour cycle. These circadian rhythms, display common properties and are generated by common cellular and molecular mechanisms. The basic rhythm is endogenously produced in cellular pacemakers in various regions; the most conspicuous are located in the central nervous system. They are composed by cells endowed with the molecular substrate necessary to generate rhythmicity, and to send the circadian time signal to the effectors of the overt rhythms. The rhythmicity in the pacemaker cells is generated by specific genes integrated in a negative feedback double loop. The structure of the genes responsible for the rhythmicity is phylogenetically well preserved. The circadian pacemakers are in turn synchronized by environmental stimuli, being light the best characterized, but other external agents, such as food and some homeostatic factors and products of metabolic activity are also effective. These synchronizing signals are conveyed to the pacemakers via specific receptors and pathways. The role of some neurotransmitters in the synchronizing action has been demonstrated. CONCLUSION: The functional integrity of this complex biochronometrical system is necessary for the maintenance of health. Its alteration results in specific ailments, and enhances the vulnerability to certain diseases PMID- 12577215 TI - [Dementia and stroke: the importance of coexisting brain pathologies]. AB - AIM: In this paper we present evidence to support the idea that vascular and degenerative pathological processes interact synergically in the genesis of post stroke dementia. METHOD: Characteristics of vascular lesions have been identified that are linked to the development of a clinical picture of dementia. Yet, the characteristics of the brain in which infarction or brain haemorrhage occur are equally important. Recent (clinical, epidemiological and neuropathological) studies highlight the high frequency with which neurodegenerative and vascular processes coexist in the same brain, and the fact that both processes share environmental and genetic risk factors. The diagnostic criteria currently in use consider the existence of a vascular pathology as being an exclusion criterion for the diagnosis of an Alzheimer type dementia, and thus rule out the possibility of studying the interaction between the two processes. Instead of restrictive criteria, pragmatic instruments must be used to classify the possible aetiology in these patients. CONCLUSIONS: It is important to study the coexistence of vascular and degenerative pathologies in the brains of patients with a post stroke deterioration. Proving the existence of a synergic relation between the two processes would open up the possibility of performing an operation to prevent and treat the dementia displayed by these patients. PMID- 12577216 TI - [Stroke of unusual origin]. AB - INTRODUCTION: Stroke of unusual origin is a heterogeneous syndrome with a number of different aetiologies. Most patients who are affected are young adults and it is often the first manifestation of a systemic disease. DEVELOPMENT: This type of stroke accounts for less than 10% of all ischemic strokes. Among its multiple aetiologies, cardioembolic and haematological diseases with a tendency toward hypocoagulability and dissections of the cervico encephalic arteries are the most common. The prognosis depends on the underlying aetiology but generally speaking this kind of stroke has a better prognosis than that of other types. CONCLUSIONS: Stroke of unusual origin is relatively rare, but nevertheless it deserves special attention since it affects young people above all, although its prognosis and treatment vary according to the aetiology. Exhaustive research needs to be conducted on these patients to establish the aetiological diagnosis and an early treatment PMID- 12577217 TI - [Repercussion of sleep apnea in neurology]. AB - INTRODUCTION: While an individual sleeps, the pneumotaxic nerve centres take over control of the breathing, instead of those that perform this function during the waking state. The respiratory disorder during sleep commonly known as sleep apnea, refers to the alterations in the breathing pattern in sleep which, when associated with diurnal drowsiness, is known as sleep apnea syndrome. METHOD: The most frequent respiratory sleep disorder is the transitory obstruction of the upper respiratory tracts, which gives rise episodes of obstructive apnea, associated with snoring, and affects 4% of the general population. The complications it can present include pharyngeal neuropathy, arterial hypertension, diurnal drowsiness, cognitive disorders and a lowered quality of life. In post stroke patients the prevalence of sleep apnea is very high and it lowers the chances of successful rehabilitation. Patients respond well to treatment of obstructive apnea with devices that supply positive pressure air and complications are relieved. CONCLUSIONS: Obstructive sleep apnea is a very widespread disorder, which can give rise to vascular complications. Patients respond well to treatment and the disease must therefore be identified and treated adequately. PMID- 12577218 TI - [Neurocysticercosis: a public health perspective]. AB - AIMS: We review the physiopathological aspects of cysticercosis that are highly relevant in public health, in particular as a cause of epilepsy in adults. Cysticercosis is the infection of the central nervous system produced by the larvae of Taenia solium. In countries where cysticercosis is endemic, the prevalence of epilepsy is twice as high as that observed in areas that are free of this parasitic disease. METHOD: The human carrier of an intestinal Taenia solium is a source of cysticercosis infection, both for others and for him or herself. We propose that all diagnosed cases of neurocysticercosis must be compulsorily reported to the department of epidemiology at the Ministry of Health, in order to enable the control of sources of infection, to search for other possibly infected subjects, as well as to determine the prevalence and geographical distribution of this problem. The aim of this would be to fit, target and optimise the available resources in eradication campaigns. CONCLUSIONS: Doctors, and more specifically neurologists and neurosurgeons, must remember that behind each patient diagnosed with neurocysticercosis there is a tapeworm carrier within the family environment or at the same table. Cysticercosis is a disease whose transmission mechanism consists in person to person contamination (from the tapeworm carrier to the individual with symptomatic or asymptomatic cysticercosis). Public health campaigns must begin from index cases of neurocysticercosis, which will allow us to reach the sources of infection. Cysticercosis is an eradicable disease that currently exists not only in developing countries but also in industrialised nations, as a result of tourism and immigration from endemic countries PMID- 12577219 TI - [Orthostatic intolerance and syncope]. AB - OBJECTIVE: To describe the current state of knowledge and clinical features of patients with othostatic intolerance. METHOD: Patients with orthostatic intolerance can be classified in two types. In one of them, sympathetic activity is diminished and they have hypotension when they stand up. This becomes chronic in patients with central or peripheral neurodegenerative diseases and is acute in those with vasovagal syncope. In the second type of orthostatic intolerance patients display an increase in sympathetic activity, without arterial hypotension and a notable tachycardia, this is called postural tachycardia syndrome. The mechanism responsible for this type of orthostatic intolerance and the exact cause of the characteristic symptoms are unknown. Diverse theories include a neuropathic lesion that affects the nerves in the lower part of the body in a selective fashion or a congenital or acquired defect in the neuronal norepinephrine transporter. CONCLUSIONS: Orthostatic intolerance is the difficulty experienced by patients to remain standing due to a dysfunction of the autonomic nervous system. Orthostatic intolerance occurs when efferent sympathetic activity is notably diminished or, paradoxically, in patients in whom efferent sympathetic activity increases in an exaggerated manner when they stand up. PMID- 12577220 TI - [Deciding on treatment in multiple sclerosis]. AB - The modern history of the treatment of multiple sclerosis is reflected during the last years with the advent of controlled clinical trials design to produce measurable outcomes utilizing MRI techniques and statistical analysis. Addressing in this form the use of disease modifying medications: immunomodulation with interferons and glatiramer acetate in the relapsing/remitting type, and combination therapies or immunosuppression in the progressive forms, these studies based on evidence provide to the clinician direction in the process to decide the most appropriate treatment for the individual patient. PMID- 12577221 TI - [Treatment of acute stroke]. AB - INTRODUCTION AND METHOD: Ischemic stroke is the first cause of invalidity and the third cause of death in modern day society. The final outcome after a stroke depends of the speed with which treatment is administered and its quality. It is for this reason that the treatment of stroke begins by applying strategies to reduce the delay in receiving medical attention both outside and inside the hospital. Care includes stabilising the patient, with special attention being given to protecting the air passages, treatment of arterial blood pressure and heartbeat rate. The initial evaluation must be quick and focused on determining a differential diagnosis, the estimation of the volume and location of the infarction, as well as considering specific therapies. Amongst the latter, the only one currently approved is thrombolysis with intravenous recombinant tissue plasminogen activator (rtPA) in selected patients and with a clinical course of less than three hours duration. Intra arterial thrombolysis is a promising therapy for patients with occlusion of the middle cerebral artery, which has been contrasted angiographically and is less than 6 hours old. CONCLUSIONS: In these moments there is no evidence to support the use of antithrombotic or neuroprotector drugs to improve the neurological state. Following the initial stabilisation and evaluation, it is advisable for the patient to be admitted to a stroke unit in order to avoid complications, to complete the evaluation and to start secondary prevention and rehabilitation. Stroke units are an intervention in themselves, since they lower mortality and invalidity. While only a minority of patients currently benefit from specific therapies, stroke units are an intervention that can potentially have positive repercussions on the huge majority of patients PMID- 12577222 TI - [The most common dysautonomias]. AB - AIMS: To review the classification and the clinical and pathological characteristics of the most common dysautonomias. METHOD: Primary dysautonomia includes neurodegenerative diseases of unknown causes that are characterised by the intracytoplasmic accumulations of alpha synuclein that manifest with four different phenotypes: pure autonomic failure, Parkinson s disease, dementia with Lewy bodies and multiple system atrophy. Of the secondary dysautonomias, diabetes mellitus is the most common cause of autonomic neuropathy in developed countries. Familiar dysautonomia is a recessive autosomic disease; the gene responsible for it has been located in the long arm of chromosome 9 (9q31). Paraneoplasic dysautonomia is associated with cancer of the lungs, the pancreas, Hodgkin's disease and testicular cancer. In Lambert Eaton myasthenic syndrome and in botulism the release of acetylcholine is deficient in both the somatic and the autonomic neurons. There are other diseases that affect autonomic cholinergic neurotransmission without bringing about any disorders in neuromuscular transmission. Chagas disease affects the neurons of the parasympathetic ganglion and produces megaesophagus, megacolon and myocardiopathy. Dopamine beta hydroxylase enzyme deficiency is a congenital disease characterised by the failure to convert dopamine into noradrenaline which results in orthostatic hypotension. CONCLUSIONS: Dysautonomias can be classified, according to their aetiology, as primary or secondary; according to the deficient neurotransmitter, as cholinergic, adrenergic and mixed (pan dysautonomia) or, according to the anatomical distribution of the neurons that are affected, as central and peripheral. PMID- 12577223 TI - [Brachial plexitis as a manifestation of giant cell arteritis]. PMID- 12577224 TI - [Neuropharmacological profile of Plectranthus amboinicus (Lour.) Spreng. (Indian borage)]. PMID- 12577225 TI - [Pyridoxine in the management of tardive dyskinesia]. PMID- 12577227 TI - Past and present treatment of pancreatic adenocarcinoma: chemotherapy as a standard treatment modality. AB - Pancreatic cancer continues to be a challenging therapeutic problem, with approximately 28,000 deaths annually in the United States. Most studies of single agent or combination chemotherapy in the treatment of patients with advanced adenocarcinoma of the pancreas have documented low response rates and little reproducible impact on patient survival or quality of life. However, the recent success of gemcitabine in patients with locally advanced and metastatic pancreatic cancer suggests that systemic therapy can have a beneficial effect on the natural history of this disease. Also, the rapidly evolving understanding of the molecular biology of pancreatic cancer may contribute to the development and use of targeted therapies with novel agents for even more effective treatments in the near future. PMID- 12577228 TI - Gemcitabine in the treatment of advanced pancreatic cancer: a comparative analysis of randomized trials. AB - In view of the data available from randomized trials, gemcitabine has been established as a new standard for the treatment of pancreatic cancer. It was shown to improve clinical benefit response, time to progression, and survival when compared with agents such as 5-fluorouracil or metalloproteinase inhibitors. In one trial, the combination of cisplatin and gemcitabine significantly improved tumor response and time to progression as compared with gemcitabine alone, while a significant impact on survival yet needs to be shown. No significant, clinically meaningful survival benefit was observed when gemcitabine was combined with bolus or infusional 5-fluorouracil, capecitabine, metalloproteinase inhibitors, or the FTI tipifarnib. Numerous ongoing randomized trials are presently investigating gemcitabine-based combination regimens involving such agents as cisplatin, oxaliplatin, irinotecan, docetaxel, 5-fluorouracil, capecitabine, or pemetrexed. PMID- 12577229 TI - Extended surgery in bilio-pancreatic cancer: the Japanese experience. AB - Aggressive surgery has been used in bilio-pancreatic cancers in many institutions in Japan. Approaches in pancreatic cancer include extended lymph node and soft tissue dissection in patients with ductal carcinoma of the pancreatic head. In biliary tract disease, approaches have included hepatectomies with or without vascular resection, reconstruction designed according to the extent of disease in individual patients, and combined liver and pancreas resection (hepatopancreatoduodenectomy) in locally advanced disease. Based on current experience, portal vein resection can be recommended in both pancreatic cancer and hilar cholangiocarcinoma and in select patients with gallbladder disease. Extended dissection of para-aortic lymph nodes is recommended in hilar cholangiocarcinoma but not in gallbladder disease; extended lymph node dissection in pancreatic cancer currently is being evaluated in a randomized controlled trial. Hepatopancreatoduodenectomy is recommended for select patients with gallbladder cancer. PMID- 12577230 TI - Role of surgery in the treatment of bilio-pancreatic cancer: the European experience. AB - Pancreatic cancer is the eighth most common cancer in Europe, accounting for 4.1% of cancer deaths in men and 4.8% in women. Standardized pancreatoduodenectomy and left pancreatectomy are the gold standard for routine surgical treatment of pancreatic cancer. Total pancreatectomy should be reserved for positive pancreatic resection margins or severe pancreatic anastomotic leak; mesenterico portal vein resection should be performed only when it is the only reasonable way to obtain clear margins. Extended lymphadenectomies are recommended only in prospective, randomized studies. Ongoing efforts to standardize surgical procedures will facilitate comparison of selection criteria and trial data among different centers, and will also optimize the study design of ongoing/planned prospective trials using adjuvant and neoadjuvant chemoradiotherapy in pancreatic cancer. PMID- 12577231 TI - Future directions in the treatment of pancreatic cancer. AB - Fewer than 20% of patients with pancreatic cancer present with disease macroscopically confined to the pancreas, and approximately 40% already have locally advanced disease. Based on data from the Gastrointestinal Tumor Study Group, adjuvant therapy with radiation and 5-fluorouracil has become standard practice in the United States; however, in other countries, adjuvant treatment has not been as widely accepted. Other issues include the potential of neoadjuvant therapy and optimal systemic management. The issue of second-line therapy has also been raised in the treatment of pancreatic carcinoma, after the establishment of gemcitabine as a first-line standard treatment approach, in which it achieves a significant clinical benefit response. Other combination partners with gemcitabine under investigation include the antimetabolite 5 fluorouracil, the topoisomerase-I inhibitor irinotecan, the taxane docetaxel, the platinum oxaliplatin, the multitargeted antifolate pemetrexed, the farnesyl transferase inhibitor R-115777, the anti-HER2/neu antibody trastuzumab, and the epidermal growth factor inhibitor cetuximab. Combined-modality approaches with gemcitabine and radiation are also under active investigation. PMID- 12577232 TI - Review of gemcitabine in biliary tract carcinoma. AB - Cancers of the biliary tract are rare, and remain a major challenge to surgical, medical, and radiation oncologists. Unfortunately, the large majority of these tumors are not resectable at the time of initial diagnosis, and patients with advanced disease face a dismal prognosis. The efficacy of conventional palliative systemic chemotherapy (eg, with 5-fluorouracil, mitomycin-C, and cisplatin) seems negligible, and there is presently no agreement on the best chemotherapeutic regimen. Gemcitabine is among several different new anticancer drugs under investigation in the treatment of advanced biliary tract cancer. Apart from its favorable toxicity profile, this novel nucleoside analog has shown activity in many solid tumors, including pancreatic adenocarcinoma. In view of the histogenetic affinity between the pancreas and the biliary tract, and several case reports describing the efficacy of gemcitabine in advanced gallbladder or cholangiocellular carcinoma, a number of phase II investigations have been undertaken. In the majority of these trials a conventional gemcitabine dose regimen of 1,000 to 1,200 mg/m(2) given over 30 minutes on 3 consecutive weeks followed by a week of rest has been used. In a total of seven studies involving 167 assessable patients, objective response rates up to 60% (36% in the largest trial composed of 39 evaluable patients), abrogation of progressive disease (complete response + partial response + stable disease) in 50% to 93%, and overall survival times ranging from 6.3 to 16 months, have been reported. The consensus is that the tolerance of treatment was remarkable with only exceptional patients (< or = 5%) experiencing grade 4 hematologic toxicities. Nonhematologic side effects were infrequent and almost exclusively mild to moderate. In three of the trials, a formal clinical benefit analysis was included, suggesting that a considerable proportion of symptomatic patients will experience relief of tumor related symptoms and/or weight gain. Possible options currently being investigated to further improve the therapeutic results of gemcitabine monotherapy include modifications of the dose regimen as well as combinations with other potentially synergistic anticancer drugs. In the latter approach, preliminary data have been reported for gemcitabine plus cisplatin, oxaliplatin, docetaxel, mitomycin-C, and continuous-infusion 5-fluorouracil/leucovorin. Objective response rates as high as 53% (for gemcitabine/cisplatin), and median survival times > or = 11 months with only a slight increase in frequency and severity of side effects have been reported. In conclusion, while the best available chemotherapeutic treatment for advanced biliary tract cancer remains to be determined, accumulating data from recent phase II trials suggest that single agent gemcitabine represents an active and very well-tolerated treatment option. It may also be safely combined with other drugs, though further improvements in response activity and survival warrant confirmation in future randomized studies. PMID- 12577233 TI - Chemotherapy in biliary tract carcinomas: results in India. AB - Epidemiology data from India suggest that biliary tract carcinomas occur primarily in the older population. Because of the absence of early symptoms, most patients present with advanced disease, and the outcome has been disappointing even in patients with resectable tumors who have undergone aggressive surgery. Although there is no standard therapy, data from recent clinical trials warrant further investigation of the role of chemotherapy in biliary tract carcinoma. In view of the favorable toxicity profile and observed efficacy of gemcitabine in advanced biliary tract cancers, its evaluation in combination with other active, potentially synergistic cytotoxic drugs such as cisplatin, capecitabine, and carboplatin should be studied with an aim to improve therapeutic efficacy and survival. PMID- 12577234 TI - Chemotherapy for biliary tract cancer in Japan. AB - The incidence of biliary tract cancer has increased markedly in Japan over the past several decades and ranks as the sixth leading cause of cancer death. Currently, there is no standard chemotherapy for biliary tract cancer. Early phase investigation of cisplatin and cisplatin/epirubicin/5-fluorouracil yielded disappointing results in this setting. Gemcitabine is currently approved for use in the treatment of non-small cell lung cancer and pancreatic cancer in Japan. Based on the single-agent activity of gemcitabine in biliary tract cancer in other locales combined with the drug's established safety profile, gemcitabine is currently being evaluated as single-agent therapy in the treatment of advanced biliary tract cancer patients in Japan. PMID- 12577235 TI - Proceedings of the 2nd International Cytoprotection Investigators' Congress. June 6-9, 2002. St Thomas, US Virgin Islands, USA. PMID- 12577237 TI - Improved tolerability of amifostine with rapid infusion and optimal patient preparation. AB - Early studies of amifostine suggested a reduced frequency of nausea/vomiting and hypotension when the drug was given via shorter-duration, intravenous infusions. Data from subsequent clinical evaluations in the radioprotectant and cytoprotectant settings have supported this observation. Recent findings indicate that amifostine given preradiotherapy by rapid intravenous push (10 seconds) and prechemotherapy over 3 to 5 minutes is associated with reduced toxicity and improved tolerability without apparent loss of protective effects. In addition to reducing duration of infusion, steps that should be taken to improve amifostine tolerability include adequate pretreatment hydration, individualized antiemetic prophylaxis, and maintaining the patient in a supine or reclining position during and after amifostine treatment. Subcutaneous dosing is currently being assessed with the aim of improving ease of administration and potentially further improving tolerability. Semin Oncol 29 (suppl 19):9-13. PMID- 12577236 TI - Preclinical studies on the radioprotective efficacy and pharmacokinetics of subcutaneously administered amifostine. AB - The radioprotective effects and pharmacokinetics of subcutaneously (SC) administered amifostine have been investigated in animal studies. Studies in rats using a single dose of amifostine showed that SC administration gave protection from radiation-induced mucositis that is at least equivalent to that achieved by intravenous administration of the drug. These studies also indicate that tissue levels of the active metabolite WR-1065 correlated better with the radioprotective effects of amifostine than do plasma WR-1065 levels. Multiple dose studies in rats show radioprotective effects equal to or greater than those obtained with intravenous dosing in the setting of fractionated irradiation. In addition, there is no evidence of drug accumulation in either normal or tumor tissue, with tumor WR-1065 levels peaking just above the limits of quantitation during treatment. Preliminary data from studies of SC amifostine in monkeys indicate a plasma pharmacokinetic profile similar to that reported earlier in humans. Tissue WR-1065 levels were higher at 30 minutes after SC dosing than they were after intravenous dosing and were comparable for the two routes at 60 minutes. PMID- 12577238 TI - Down-regulation of intestinal-type alkaline phosphatase in the tumor vasculature and stroma provides a strong basis for explaining amifostine selectivity. AB - Strong clinical and experimental evidence indicates that amifostine confers cytoprotection in normal, but not in tumor, tissues. However, the mechanism of such selective action is poorly understood. Intestinal-type alkaline phosphatase (IAP) is a major isoenzyme involved in the hydrolysis of amifostine (WR-2721) to its active thiol form WR-I065. Could differences in IAP expression between normal and malignant tissues account for amifostine's selectivity? Paraffin-embedded material from normal breast, lung, colon, and head and neck tissues, together with their malignant counterparts, were retrieved and stained immunohistochemically for human IAP (antibody 7324, Abcam, Cambridge, UK) and endothelial cell CD31 antigen. Normal tissues (epithelium, fibroblasts, and vessels) consistently displayed strong nuclear and cytoplasmic IAP reactivity. The vascular density (number of positive vessels per x 200 optical field), whether assessed in anti-IAP or anti-CD31 stained sections, was very similar, indicating a strong IAP content for the entire normal vasculature. Therefore, amifostine hydrolysis is ensured in normal tissues and may occur at both vascular and interstitial levels. By contrast, 60% of the tumors analyzed showed a loss of IAP expression in both epithelial cells and stroma, and only 10% to 15% of them demonstrated nuclear/cytoplasmic reactivity, which was confined to the epithelial cells. Similarly, the percentage of tumor vessels exhibiting some IAP reactivity was very low (6% to 17%). This dramatic loss of IAP expression from tumor stroma/vasculature may form a strong basis for explaining amifostine selectivity. In contrast, the abundance of IAP expression in normal tissues, stromal and vascular, ensures an intense hydrolysis of WR-2721 and rapid intracellular accumulation of WR-1065. PMID- 12577239 TI - Antimetastatic effectiveness of amifostine therapy following surgical removal of Sa-NH tumors in mice. AB - The effects of dose per fraction on the ability of amifostine exposure to elevate angiostatin levels in the serum of mice and to inhibit spontaneous metastases formation using the well-characterized murine Sa-NH sarcoma were investigated. Amifostine was administered intraperitoneally at doses of 50, 100, or 200 mg/kg every other day for 6 days to C3Hf/Kam mice until tumors reached an average size of 8 mm in diameter. Amifostine was again administered immediately following surgical removal of the tumor-bearing limbs by amputation, and then once more 2 days later. Nontumor-bearing control animals were treated using the same dosing and surgery schedules. The average number of pulmonary metastases per animal was determined for each experimental group. A significant reduction (P <.05) in the average number of pulmonary metastases was observed only in the group of animals exposed to a dose per fraction of 50 mg/kg. A dose of 100 mg/kg was less effective while 200 mg/kg had no effect on metastases formation in this study. The effects of amifostine exposure on serum levels of the angiogenesis inhibitor angiostatin were also determined using Western analysis. Correlating with the antimetastatic effect measured, exposure of animals to 50 mg/kg of amifostine resulted in a four-fold enhanced serum level of angiostatin above control levels. This phenomenon occurred in both tumor-bearing as well as nontumor-bearing animals. In contrast, a dose of 200-mg/kg amifostine administered intraperitoneally under these conditions had no measurable effect on angiostatin serum levels in this animal system. The enhanced ability of relatively low doses of amifostine to inhibit spontaneous metastases formation suggests that effective antimetastatic therapies with amifostine can be designed with minimal toxic side effects. While the dose responses for angiostatin production and metastases inhibition by amifostine are well correlated, the precise mechanism of action underlying these phenomena is unclear but is suggestive of a redox driven process(es). PMID- 12577240 TI - Early results from a phase I/II radiation dose-escalation study with concurrent amifostine and infusional 5-fluorouracil chemotherapy for preoperative treatment of unresectable or locally recurrent rectal carcinoma. AB - To improve the resectability and long-term local control of locally advanced rectal cancer, we have initiated a radiation dose-escalation trial. It is hoped that the radioprotector amifostine will sufficiently reduce rectal morbidity (without also reducing tumor response) to permit increasing radiation doses. Only 6 patients have been treated at the first dose level (45 Gy with a 9-Gy boost and concurrent continuous infusion of 5-fluorouracil). During treatment, daily stool frequency improved or was stable. There was one case of transitory severe proctitis/enteritis. Four of the six patients achieved objective responses to preoperative treatment. These very early results suggest that the use of amifostine does not interfere with tumor response and that it may facilitate the delivery of higher boost doses to the rectum. PMID- 12577241 TI - Potential for use of amifostine in cervical cancer. AB - Cisplatin-based combined chemoradiotherapy is now the standard of treatment for locally advanced cervical cancer. Although combined-modality treatment has improved survival, it has also increased the already high frequency of toxicity observed with traditional radiotherapy alone. Cervical cancer patients with para aortic metastasis may be at particularly high risk of toxicity with combined modality treatment. The few clinical trial data available on the use of amifostine in cervical cancer or pelvic cancer patients suggest benefit in reducing treatment toxicity. The potential role of amifostine in combination with currently used cisplatin-based chemoradiotherapy regimens is being assessed in a Radiation Therapy Oncology Group phase I/II study in cervical cancer patients with positive para-aortic or high common iliac lymph nodes. PMID- 12577242 TI - A phase III randomized evaluation of amifostine in stage IIIA/IIIB non-small cell lung cancer patients receiving concurrent carboplatin, paclitaxel, and radiation therapy followed by gemcitabine and cisplatin intensification: preliminary findings. AB - Despite the progress in pharmaceutical therapeutics and the hardware and software advances supporting radiation oncology, the major advance in the treatment of locally advanced nonmetastatic non-small cell lung cancer during the previous decade has been the adoption of integrated combined-modality therapy. This has resulted in an appreciation of the associated increase in treatment-related morbidity that has fostered new efforts to maximize the therapeutic index, including evaluation of cytoprotective agents. This report presents the preliminary data from a prospective randomized phase III trial of radiation therapy (64.8 Gy) administered concurrently with paclitaxel and carboplatin with posterior consolidation with cisplatin and gemcitabine with or without amifostine. The data to date do not support a cytoprotective advantage to the use of intravenous amifostine targeting esophagitis at 200 mg/m2/d before radiation. Further efforts to assess the role and potential of amifostine in the treatment of patients with non-small cell lung cancer are recommended. PMID- 12577243 TI - Hypofractionated and accelerated radiotherapy with amifostine cytoprotection (HypoARC): a new concept in radiotherapy and encouraging results in breast cancer. AB - Tumors with low intrinsic radiosensitivity, low alpha/beta-ratios and, eventually, hypoxic tumors unable to undergo reoxygenation (because of intrinsic reasons or radiotherapy acceleration that delivers the total dose before the onset of reoxygenation) are likely to respond better to large fractions of radiotherapy. Accelerating the course of hypofractionated radiotherapy would further target tumors with high repopulation ability. Such hypofractionated and accelerated schedules are expected to confer unacceptable toxicity. Accordingly, we investigated the cytoprotective efficacy of amifostine as a potential key to permit the safe administration of such highly toxic regimens. Hypofractionated and accelerated radiotherapy with amifostine cytoprotection (HypoARC; 3.5 to 4 Gy/d x 12 days) was tested in phase I and II trials in patients with breast cancer. Early radiation skin toxicity was significantly lower, and late toxicity from breast, lung, muscle/bone structures, and nerves compared favorably with that associated with standard radiotherapy. The reduced overall treatment time (16 days v 44 days) renders HypoARC appealing, especially to older patients or patients residing some distance from radiotherapy centers. Molecular studies showed that HypoARC significantly improves local relapse-free survival in patients with highly proliferating tumors or tumors overexpressing proteins linked to increased resistance to radiation-induced apoptosis (ie, c-erbB-2 and mutant p53). Further clinical trials have been designed to test the safety and efficacy of HypoARC in other tumor types. PMID- 12577244 TI - Rationale for a phase III study of erythropoietin as a neurocognitive protectant in patients with lung cancer receiving prophylactic cranial irradiation. AB - Recent meta-analysis of patients with small cell lung cancer has confirmed the effectiveness of prophylactic cranial irradiation in reducing the cumulative incidence of brain metastases and contributing to a significant increase in 3 year survival. Likewise, with increased median survivals being documented in patients with stage IIIA/B non-small cell lung cancer, there is evidence that the brain is emerging as a significant metastatic target site. Although prophylactic cranial irradiation is a reasonable option to explore, the potential for long term neuropsychologic adverse effects is of concern in both diagnostic groups. Radiation-induced reactive oxygen intermediates and reactive nitrogen intermediates appear to play a major role in mediating this toxicity. Hypoxic stress results in a significant increase in erythropoietin (EPO) mRNA in mouse brain and, in two models, the administration of EPO improves performance function and prevents cognitive impairment. With the demonstration of EPO receptors in astrocytes, neurons, and brain capillary endothelial cells as well as the ability of EPO to cross the blood-brain barrier, a potential for EPO-mediated central nervous system radioprotection is postulated. The rationale and preliminary design for a phase III study of EPO as a neurocognitive protectant in patients with lung cancer receiving prophylactic cranial irradiation is presented. PMID- 12577245 TI - The potential of amifostine in high-dose chemotherapy and autologous hematopoietic stem cell transplantation. AB - Amifostine treatment may allow chemotherapy dose increases beyond those permitted by autologous hematopoietic stem cell transplantation. In a recent study in patients with solid tumors receiving a high-dose regimen of etoposide, ifosfamide, and carboplatin plus autologous stem cell transplantation, amifostine pretreatment was associated with significant reductions in time to neutrophil and thrombocyte engraftment, fewer days of neutropenic fever, less need for salvage antibiotic therapy. Also, there were significant reductions in grade 3 or 4 stomatitis/diarrhea, and delayed nausea/vomiting. A phase I/II study in patients with refractory/high-risk malignancies indicated that a 140% increase of high dose melphalan (up to 280 mg/m2) can be safely used with amifostine and autologous stem cell transplantation with manageable mucosal toxicity and a reduced incidence of regimen-related toxicity. Preliminary findings in another phase II study indicate that melphalan 280 mg/m2 can also be safely used with amifostine/stem cell transplantation in the treatment of patients with myeloma. Additional studies are ongoing or planned to examine the potential hematoprotective and hematostimulating effects of amifostine in the setting of high-dose chemotherapy and autologous stem cell transplantation. PMID- 12577246 TI - Preliminary data of the GORTEC 2000-02 phase III trial comparing intravenous and subcutaneous administration of amifostine for head and neck tumors treated by external radiotherapy. AB - A prospective randomized study is comparing intravenous (IV; arm A) versus subcutaneous (SC; arm B) administration of amifostine in patients receiving radiotherapy for head and neck cancer. Main eligibility criteria were newly diagnosed squamous cell head and neck cancer, inclusion of at least 75% of both parotid glands within radiation fields that would receive at least 40 Gy, and no evidence of distant metastasis. Prophylactic use of pilocarpine and concomitant chemotherapy were prohibited. Intravenous administration of amifostine is 200 mg/m2/d in a short 3-minute infusion 15 to 30 minutes before each fraction of radiotherapy. Subcutaneous administration is 500 mg/d in two, slow 1.25-mL injections at two different sites 20 to 60 minutes before each radiotherapy fraction. Antiemetic treatment and blood pressure monitoring are required in both arms. As of April 25, 2002, 111 of the 292 required patients were included. Data are available for the first 54 patients. Acute toxicity included nausea/vomiting (12% for arm A; 13% for arm B), hypotension (6% in arm A; 0% in arm B), skin rash (15% in arm A; 16% in arm B), and asthenia (4% in arm A; 0% in arm B). Compliance with amifostine administration was 70% in arm A (IV) and 80% in arm B (SC). The rate of acute xerostomia (> or = grade 2) was 23% in arm A and 19% in arm B. These preliminary results indicate that tolerance is better with SC than IV administration, particularly because of the absence of hypotension. The absence of hypotension with SC administration facilitates patient monitoring and management in radiotherapy departments. More patients and data are required to assess the long-term efficacy of SC administration on acute and late xerostomia. PMID- 12577247 TI - Radioprotective effect of amifostine in patients with head and neck squamous cell carcinoma. AB - Amifostine (Ethyol; MedImmune Oncology, Gaithersburg, MD) is a radio- and chemoprotective agent currently in clinical use. Based on experimental data showing the potential for mucosal protection and additional information in animal studies showing rapid uptake of amifostine in the salivary glands along with effective radioprotection, several investigators have tested this drug in patients with head and neck squamous cell carcinoma (HNSCC) undergoing radiotherapy. The first evidence that amifostine, administered at 200 mg/m(2)/d intravenously before each radiotherapy session, could effectively protect salivary function in patients with HNSCC was provided by McDonald et al in a limited series of HNSCC patients. On the basis of these phase I results, an international multicenter phase III radiotherapy trial with or without amifostine was carried out by Brizel et al in a series of 315 patients with head and neck tumors showing a reduction in xerostomia with no suggestion of tumor protection. Additional data have been obtained in patients with HNSCC regarding the potential protective effect of amifostine on the duration and severity of radio-induced mucositis. This effect was reported in small, randomized studies of patients receiving intensive accelerated radiotherapy (Bourhis et al) or combined radiochemotherapy (Buntzel et al). In conclusion, the experience obtained to date in HNSCC patients treated with radiotherapy supports the selective cytoprotective activity of amifostine to minimize radiation effects while apparently not diminishing tumor control. PMID- 12577248 TI - Tools for optimal tissue sparing in concomitant chemoradiation of advanced head and neck cancer: subcutaneous amifostine and computed tomography-based target delineation. AB - Organ preservation in patients with head and neck cancer can be achieved using concomitant chemoradiation protocols. Critical tissues can be spared using highly conformal radiation therapy techniques and/or radiation protectors. With three dimensional conformal radiation therapy (3DCRT) tight target definitions of the primary tumor and neck nodal levels are mandatory. In 2000, a clinical trial for advanced-stage head and neck squamous cell carcinoma was initiated in Rotterdam, The Netherlands. Patients are treated with paclitaxel administered concomitantly with 3DCRT and randomized to receive subcutaneous (SC) amifostine or no amifostine. Those in the radioprotectant arm received amifostine 500 mg SC before each radiation therapy (RT) fraction. This article presents early findings on toxicity. Acute toxicity is evaluated according to Radiation Therapy Oncology Group criteria. Xerostomia was scored subjectively and by whole saliva measurements. Neck nodal levels were delineated in accordance with previously published computed tomography (CT)-based guidelines developed in Rotterdam. Forty one patients are the subject of this report. In patients treated with amifostine, mucositis and dysphagia took longer to resolve than with conventional RT schedules. No difference in objective and subjective evaluation of xerostomia was seen between treatment arms. So far in this ongoing study, no advantage of SC amifostine has been detected. This might be because of the toxicity of the concomitant treatment itself, the dose of amifostine, the route of administration, or the insufficient sparing of critical structures by 3DCRT. These early findings and the ongoing development of better tissue-sparing techniques with more accurate CT-based target delineation protocols and intensity modulated radiation therapy (IMRT) are discussed. PMID- 12577249 TI - Using technology to decrease xerostomia for head and neck cancer patients treated with radiation therapy. AB - The treatment of head and neck cancer has evolved from conventional fields encompassing large volumes of normal tissue to focused treatment aimed at conforming the dose around the target while avoiding normal tissue. Intensity modulated radiation therapy has changed the way radiation oncologists think about head and neck cancer. Using the concepts of conformal treatment and avoidance, the therapeutic ratio can be improved and technology exploited to the patients' advantage. This is particularly evident with head and neck irradiation, where a common side effect is xerostomia. By decreasing xerostomia through conformal avoidance of the parotid glands, we can improve patient satisfaction and quality of life. In this study, xerostomia is assessed through a subjective salivary gland function questionnaire. This article examines the use of intensity modulated radiation therapy in the treatment of head and neck cancer to decrease xerostomia. The purpose of this article is to evaluate the significance of parotid gland dosimetry in relation to subjective salivary gland function. PMID- 12577250 TI - Phase II trial of subcutaneous amifostine in patients undergoing radiation therapy for head and neck cancer. AB - The availability of subcutaneously (SC) administered amifostine may present an advantage for radioprotectant therapy in head and neck cancer patients. In a randomized phase II trial comparing SC amifostine versus no amifostine in 140 patients undergoing radiation therapy for head and neck, thoracic, or pelvic cancers, amifostine treatment was associated with reductions in mucosal toxicity and delays in radiation therapy among the 19 patients with head and neck cancer, as well as in the thoracic and pelvic cancer groups. A phase II trial of SC amifostine in head and neck cancer was performed in a patient population (n = 54) similar to that studied in a phase III trial of intravenous amifostine to allow comparisons of outcomes. Acute xerostomia grade 2 occurred in 56% with SC amifostine and 51% with intravenous amifostine (78% in the no-amifostine group in phase III trial), with median time to onset being 40 days and 45 days, respectively (30 days with no amifostine), and cumulative radiation dose to onset being 58 Gy and 60 Gy (42 Gy with no amifostine), respectively. Amifostine SC was well tolerated, with three quarters of patients receiving > or =75% of the planned dose. Nausea, vomiting, and hypotension were less severe with SC amifostine, but cutaneous toxicity was more frequent. The reduction in radiation therapy-induced acute xerostomia with SC amifostine is similar to that with intravenous amifostine in patients with head and neck cancer. If cutaneous toxicity is judged an acceptable risk, SC amifostine may represent a second, more convenient option for treating physicians. PMID- 12577252 TI - Understanding the impact of painful diabetic neuropathy. AB - Painful neuropathy is a common and often distressing complication of diabetes. It has considerable impact on the social and psychological well-being of affected individuals. There are two distinct forms of painful neuropathy: an acute and self-limiting form that resolves within a year or a chronic form that can go on for years. There are now a number of drugs available for the treatment of neuropathic pain. However, some may fail to respond to these drugs or may have unacceptable adverse side effects. When this is the case, the patient's quality of life can be severely affected. Health care professionals need to assess the full impact of painful neuropathy. In this article we review a number of instruments that are used to assess the severity of painful neuropathy and its impact on the quality of life. PMID- 12577253 TI - Neuropathic pain and diabetes. AB - Neuropathic pain is a common phenomenon resulting from injury to the central or peripheral nervous system. The means by which diabetes results in nerve injury is unclear but the effect is to cause injury at all levels of the nervous system from the level of the peripheral nerves to the brain. Nerve injury causes pain through a cascade of mechanisms resulting in altered processing of sensory input into the nervous system. This alteration occurs through chemical and anatomical changes in the nervous system that are similar to some of the processes seen in central sensitisation following acute pain. Following nerve injury, neuropathic pain occurs not only when these mechanisms are activated but also when sensitisation is maintained. Other processes occurring in neuropathic pain appear to be a loss of normal inhibitory controls as seen by a reduction in local GABA ergic and descending monoaminergic influences. There are also important changes mediated via glial cells that can maintain neuropathic pain. Diabetes affects all areas of the nervous system and the contribution of higher levels of the nervous system is often overlooked. Neurophysiological and MRI evidence strongly suggest that these may contribute to the pain of diabetic neuropathy. Psychological dysfunction in diabetic patients is an important factor in increasing the suffering associated with all aspects of the disease, but treatment and control of pain can greatly improve the quality of life. PMID- 12577254 TI - Treatment of symptomatic diabetic neuropathy. AB - Painful diabetic neuropathy is a common and particularly unpleasant long-term complication of diabetes that affects a significant minority of patients with distal polyneuropathy. After exclusion of other causes of neuropathic pain, attention should be focused on achieving optimal and stable glycaemic control avoiding flux of blood glucose levels, which have been shown to aggravate pain. Most patients will require pain control therapy and whilst the tricyclic drugs remain a first-line approach, their use is often hampered by predictable but troublesome side effects. Gabapentin, the only agent specifically licensed for the treatment of neuropathic pain in the United Kingdom, is useful in diabetic neuropathy and is generally better tolerated than the tricyclics. Additionally, other pharmacological and non-pharmacological pain management approaches may be useful. Patient education has a significant role to play in the avoidance of late neurological complications. PMID- 12577255 TI - Painful diabetic neuropathy--providing the best patient service. AB - Painful diabetic neuropathy is just one of a myriad of secondary conditions that may result from poor glycaemic control. Educating the patient to suspect diabetic complications, understanding why they may occur, what their progression is likely to be and what measures can be taken to avoid them can significantly improve outcomes. Education within the diabetes management framework needs to be consistent, prioritised, tailored to the needs of each patient and ongoing in order to be maximally effective, with tight glycaemic control playing the anchoring central role. The specialist diabetes nurse is in the ideal situation to coordinate this education and to ensure that the patient with diabetes receives the care and support needed to take dual responsibility for their condition. PMID- 12577257 TI - Crystal structure of a hypothetical protein, TM841 of Thermotoga maritima, reveals its function as a fatty acid-binding protein. AB - We determined the three-dimensional (3D) crystal structure of protein TM841, a protein product from a hypothetical open-reading frame in the genome of the hyperthermophile bacterium Thermotoga maritima, to 2.0 A resolution. The protein belongs to a large protein family, DegV or COG1307 of unknown function. The 35 kDa protein consists of two separate domains, with low-level structural resemblance to domains from other proteins with known 3D structures. These structural homologies, however, provided no clues for the function of TM841. But the electron density maps revealed clear density for a bound fatty-acid molecule in a pocket between the two protein domains. The structure indicates that TM841 has the molecular function of fatty-acid binding and may play a role in the cellular functions of fatty acid transport or metabolism. PMID- 12577258 TI - Fine-grained protein fold assignment by support vector machines using generalized npeptide coding schemes and jury voting from multiple-parameter sets. AB - In the coarse-grained fold assignment of major protein classes, such as all alpha, all-beta, alpha + beta, alpha/beta proteins, one can easily achieve high prediction accuracy from primary amino acid sequences. However, the fine-grained assignment of folds, such as those defined in the Structural Classification of Proteins (SCOP) database, presents a challenge due to the larger amount of folds available. Recent study yielded reasonable prediction accuracy of 56.0% on an independent set of 27 most populated folds. In this communication, we apply the support vector machine (SVM) method, using a combination of protein descriptors based on the properties derived from the composition of n-peptide and jury voting, to the fine-grained fold prediction, and are able to achieve an overall prediction accuracy of 69.6% on the same independent set-significantly higher than the previous results. On 10-fold cross-validation, we obtained a prediction accuracy of 65.3%. Our results show that SVM coupled with suitable global sequence-coding schemes can significantly improve the fine-grained fold prediction. Our approach should be useful in structure prediction and modeling. PMID- 12577259 TI - Folding in lipid membranes (FILM): a novel method for the prediction of small membrane protein 3D structures. AB - We present the results of applying a novel knowledge-based method (FILM) to the prediction of small membrane protein structures. The basis of the method is the addition of a membrane potential to the energy terms (pairwise, solvation, steric, and hydrogen bonding) of a previously developed ab initio technique for the prediction of tertiary structure of globular proteins (FRAGFOLD). The method is based on the assembly of supersecondary structural fragments taken from a library of highly resolved protein structures using a standard simulated annealing algorithm. The membrane potential has been derived by the statistical analysis of a data set made of 640 transmembrane helices with experimentally defined topology and belonging to 133 proteins extracted from the SWISS-PROT database. Results obtained by applying the method to small membrane proteins of known 3D structure show that the method is able to predict, at a reasonable accuracy level, both the helix topology and the conformations of these proteins. PMID- 12577260 TI - Observation of residual dipolar couplings in short peptides. AB - Residual dipolar couplings provide information on the orientation of individual bond vectors with respect to a unique set of molecular axes. We report that short peptides from 2 to 15 amino acids in length of arbitrary sequence exhibit a modest range of residual dipolar couplings when aligned in either strained polyacrylamide gels or alkyl-PEG bicelles. The absence of significant line broadening in gels suggests peptides align predominantly through steric interactions with the polyacrylamide matrix. However, broadening of NMR lines for a subset of residues aligned in bicelles indicates some peptides bind weakly to these lipid disks, yet a weak negative correlation between the couplings measured in gels and bicelles is consistent with steric hindrance playing a role in both media. The observation of dipolar couplings for peptides of length 10-15 suggests the statistical segment lengths of polypeptide chains must often be >10-15 residues, with data from denatured proteins indicating even larger values. Presumably, local side-chain backbone interactions severely restrict chain flexibility, with the cumulative effect of many such restrictions giving rise to biases in chain direction that may persist for the entire length of a protein chain. Comparison of experimental dipolar couplings for peptides with couplings calculated for ensembles of conformations generated by molecular dynamics should permit evaluation of the accuracy of molecular mechanics potentials in reproducing sequence-specific preferences for phi and psi angles. PMID- 12577261 TI - Helix propensities of short peptides: molecular dynamics versus bioinformatics. AB - Knowledge-based potential functions for protein structure prediction assume that the frequency of occurrence of a given structure or a contact in the protein database is a measure of its free energy. Here, we put this assumption to test by comparing the results obtained from sequence-structure cluster analysis with those obtained from long all-atom molecular dynamics simulations. Sixty-four eight-residue peptide sequences with varying degrees of similarity to the canonical sequence pattern for amphipathic helix were drawn from known protein structures, regardless of whether they were helical in the protein. Each was simulated using AMBER6.0 for at least 10 ns using explicit waters. The total simulation time was 1176 ns. The resulting trajectories were tested for reproducibility, and the helical content was measured. Natural peptides whose sequences matched the amphipathic helix motif with greater than 50% confidence were significantly more likely to form helix during the course of the simulation than peptides with lower confidence scores. The sequence pattern derived from the simulation data closely resembles the motif pattern derived from the database cluster analysis. The difficulties encountered in sampling conformational space and sequence space simultaneously are discussed. PMID- 12577262 TI - D-amino acid residues in peptides and proteins. AB - We have investigated the D-amino acid residues present in Protein Data Bank (PDB) entries, categorizing them into "real" D-residues and artifacts. In polypeptide chains of more than 20 residues, only a single instance of a "real" D-residue, other than those deliberately designed or engineered, was found. This example was the result of a slow chemical epimerization process. Another 12 designed D residues were found in these longer polypeptide chains. Smaller peptides of 20 or fewer residues contained 479 "real" D-residues, the majority in various gramicidin, actinomycin, or cyclosporin structures. We found 148 PDB entries with "real" D-residues and a further 186, in which all apparent D-residues are artifacts. Investigating the (phi, psi) preferences of the "real" D-residues, we found that the region around (-60 degrees, -45 degrees ) was almost completely unoccupied, even though it is not formally disallowed. We link the low propensity to occupy this region with the alpha-helix destabilizing properties of D residues. PMID- 12577263 TI - Protein local structure prediction from sequence. AB - A basis set of protein canonical fragments, or centroids, represents the range of local structure found in globular proteins. We develop a methodology to predict centroids from the amino acid sequence. The predictor gives the probability of each centroid in the basis set, at each loci along the backbone. The predictor selects the best-fit centroid at about 40% of the loci. The predicted probabilities are accurate and can be used to judge the confidence of each centroid prediction. For example, when filtering out centroids with <0.50 probability, the predictor is 65% accurate, although such high-probability centroids occur at only 28% of the loci. Centroids with high probability can be interpreted as segments that are highly influenced by the amino acid sequence, whereas centroids with low probability can be interpreted as segments that are more likely influenced by tertiary contacts. Low-resolution, starting point structures, can be generated by fitting the predicted centroids together. PMID- 12577264 TI - Protein fragment clustering and canonical local shapes. AB - A novel clustering method is used to cluster protein fragments by shape. The centroids (mean fragments from each cluster) form a basis set of structural motifs. A database of 156,643 seven-residue fragments is used, and eight different basis sets with varying levels of resolution are generated. Coarse basis sets contain tens of centroids and provide meaningful local shapes, which are more detailed than the traditional secondary structure categories. High resolution basis sets contain thousands of centroids and can be used to model tertiary structure of longer segments. The basis sets generated fit nontraining set proteins with the expected accuracy. PMID- 12577265 TI - A path from primary protein sequence to ligand recognition. AB - A novel method to organize protein structural information based solely on sequence is presented. The method clusters proteins into families that correlate with the three-dimensional protein structure and the conformation of the bound ligands. This procedure was applied to nicotinamide adenine dinucleotide [NAD(P)] utilizing enzymes to identify a total of 94 sequence families, 53 of which are structurally characterized. Each of the structurally characterized proteins within a sequence family correlates to a single protein fold and to a common bound conformation of NAD(P). A wide range of structural folds is identified that recognize NAD(P), including Rossmann folds and beta/alpha barrels. The defined sequence families can be used to identify the type and prevalence of NAD(P) utilizing enzymes in the proteomes of sequenced organisms. The proteome of Mycobacterium tuberculosis was mined to generate a proteome-wide profile of NAD(P)-utilizing enzymes coded by this organism. This enzyme family comprises approximately 6% of the open reading frames, with the largest subgroup being the Rossmann fold, short-chain dehydrogenases. The preponderance of short-chain dehydrogenases correlates strongly with the phenotype of M. tuberculosis, which is characterized as having one of the most complex prokaryotic cell walls. PMID- 12577266 TI - A neural network approach to evaluate fold recognition results. AB - Fold recognition techniques assist the exploration of protein structures, and web based servers are part of the standard set of tools used in the analysis of biochemical problems. Despite their success, current methods are only able to predict the correct fold in a relatively small number of cases. We propose an approach that improves the selection of correct folds from among the results of two methods implemented as web servers (SAMT99 and 3DPSSM). Our approach is based on the training of a system of neural networks with models generated by the servers and a set of associated characteristics such as the quality of the sequence-structure alignment, distribution of sequence features (sequence conserved positions and apolar residues), and compactness of the resulting models. Our results show that it is possible to detect adequate folds to model 80% of the sequences with a high level of confidence. The improvements achieved by taking into account sequence characteristics open the door to future improvements by directly including such factors in the step of model generation. This approach has been implemented as an automatic system LIBELLULA, available as a public web server at http://www.pdg.cnb.uam.es/servers/libellula.html. PMID- 12577267 TI - The role of electrostatic interaction in the molecular recognition of selective agonists to metabotropic glutamate receptors. AB - The influence of electrostatic interactions in determining selectivity for individual subtypes of metabotropic glutamate receptors (mGluRs) is evaluated for a small set of agonists by using the program Delphi and the information thus obtained is compared with docking experiments carried out with AutoDock. The evaluation of the electrostatic component of the free energy of binding for L Glu, L-AP4, or S-PPG to mGluR1, mGluR2, and mGluR4 subtypes allowed for the detection of subtle differences in the electronic properties of the three subtypes, differences that can account for the observed agonist selectivity. PMID- 12577268 TI - Solubilization of cellulosomal cellulases by fusion with cellulose-binding domain of noncellulosomal cellulase engd from Clostridium cellulovorans. AB - Clostridium cellulovorans produces a cellulase complex (cellulosome) as well as noncellulosomal cellulases. In this study, we determined a factor that affected the solubility of the cellulosomal cellulase EngB and the noncellulosomal EngD when they were expressed in Escherichia coli. The catalytic domains of EngB and EngD formed inclusion bodies when expressed in E. coli. On the other hand, both catalytic domains containing the C-terminal cellulose-binding domain (CBD) of EngD were expressed in soluble form. Fusion with the CBD of EngD also helped increased the solubility of cellulosomal cellulase EngL upon expression in E. coli. These results indicate that the CBD of EngD plays an important role in the soluble expression of the catalytic domains of EngB, EngL, and EngD. The possible mechanisms of solubilization by fusion of the catalytic domain with the CBD from EngD are discussed. PMID- 12577269 TI - Real value prediction of solvent accessibility from amino acid sequence. AB - The solvent accessibility of amino acid residues has been predicted in the past by classifying them into exposure states with varying thresholds. This classification provides a wide range of values for the accessible surface area (ASA) within which a residue may fall. Thus far, no attempt has been made to predict real values of ASA from the sequence information without a priori classification into exposure states. Here, we present a new method with which to predict real value ASAs for residues, based on neighborhood information. Our real value prediction neural network could estimate the ASA for four different nonhomologous, nonredundant data sets of varying size, with 18.0-19.5% mean absolute error, defined as per residue absolute difference between the predicted and experimental values of relative ASA. Correlation between the predicted and experimental values ranged from 0.47 to 0.50. It was observed that the ASA of a residue could be predicted within a 23.7% mean absolute error, even when no information about its neighbors is included. Prediction of real values answers the issue of arbitrary choice of ASA state thresholds, and carries more information than category prediction. Prediction error for each residue type strongly correlates with the variability in its experimental ASA values. PMID- 12577271 TI - Thermal unfolding simulations of apo-calmodulin using leap-dynamics. AB - The simulation method leap-dynamics (LD) has been applied to protein thermal unfolding simulations to investigate domain-specific unfolding behavior. Thermal unfolding simulations of the 148-residue protein apo-calmodulin with implicit solvent were performed at temperatures 290 K, 325 K, and 360 K and compared with the corresponding molecular dynamics trajectories in terms of a number of calculated conformational parameters. The main experimental results of unfolding are reproduced in showing the lower stability of the C-domain: at 290 K, both the N- and C-domains are essentially stable; at 325 K, the C-domain unfolds, whereas the N-domain remains folded; and at 360 K, both domains unfold extensively. This behavior could not be reproduced by molecular dynamics simulations alone under the same conditions. These results show an encouraging degree of convergence between experiment and LD simulation. The simulations are able to describe the overall plasticity of the apo-calmodulin structure and to reveal details such as reversible folding/unfolding events within single helices. The results show that by using the combined application of a fast and efficient sampling routine with a detailed molecular dynamics force field, unfolding simulations of proteins at atomic resolution are within the scope of current computational power. PMID- 12577270 TI - Crystal structures of a psychrophilic metalloprotease reveal new insights into catalysis by cold-adapted proteases. AB - Enzymes from psychrophilic organisms differ from their mesophilic counterparts in having a lower thermostability and a higher specific activity at low and moderate temperatures. It is in general accepted that psychrophilic enzymes are more flexible to allow easy accommodation and transformation of the substrates at low energy costs. Here, we report the structures of two crystal forms of the alkaline protease from an Antarctic Pseudomonas species (PAP), solved to 2.1- and 1.96-A resolution, respectively. Comparative studies of PAP structures with mesophilic counterparts show that the overall structures are similar but that the conformation of the substrate-free active site in PAP resembles that of the substrate-bound region of the mesophilic homolog, with both an active-site tyrosine and a substrate-binding loop displaying a conformation as in the substrate-bound form of the mesophilic proteases. Further, a region in the catalytic domain of PAP undergoes a conformational change with a loop movement as large as 13 A, induced by the binding of an extra calcium ion. Finally, the active site is more accessible due to deletions occurring in surrounding loop regions. PMID- 12577272 TI - Evidence that the SSRI dose response in treating major depression should be reassessed: a meta-analysis. AB - The limitations in design and analysis of currently available dose-response studies of SSRI treatment of major depression have led to the conclusion that dose response is flat. We applied concepts from our companion article to determine if currently available data is consistent with a "potential" and an "expressed" dose response. Using these concepts, we performed a meta-analysis on all identifiable published fixed-dose and dose-escalation studies that reported the effect of different SSRI oral doses on efficacy. "Potential" dose response in fixed-dose studies with categorical response outcomes equaled a significant meta analyzed slope of 3.1%/100 SSRI mg equivalents (SMEs) (SE=1.2%) or 7.8% across the dose range. Similar analysis in dose-escalation studies that reported categorical response data yielded a non-significant meta-analyzed slope of 3.7%/100 SMEs (SE=2.3%) or 9.3% across the dose range. Analyses of the "expressed" dose response demonstrated in the studies indicated a slope statistically equal to zero. The current analysis suggests a "potential" dose response can be demonstrated for SSRIs in treating major depression. The analysis suggests an "expressed" dose response could exist in best clinical practice. Study designs better tailored to address the relevant clinical question would test these hypotheses more appropriately than previous studies. PMID- 12577273 TI - Is there a SSRI dose response in treating major depression? The case for re analysis of current data and for enhancing future study design. AB - It has been widely stated that the available research data has not demonstrated a SSRI dose response for major depression. We re-evaluated the methods used to analyze the SSRI data by clarifying two key alternative definitions of dose response and their implications for enhancing analysis of currently available data as well as future study design. We differentiated "potential" dose response, which focuses exclusively on response excluding tolerability effects and asks whether differences in dose can result in significant differences in response, from "expressed" dose response, which incorporates all tolerability effects currently associated with dose (including those caused by study protocol or treatment practice) and asks whether differences in dose do result in significant differences in response. To analyze potential dose response for all studies, one should use a "dose-tolerant" sample, i.e., an ITT sample from which dropouts due to adverse events have been removed. To analyze an expressed dose response, an ITT sample is the optimum sample if the study conforms to several design specifications. In the absence of conformance to these specifications, an ITT sample may be an approximation of the appropriate sample. Given design limitations of currently available studies, a dose-tolerant sample may provide a more informative approximation of an optimal sample to be used in evaluating the expressed dose response that could be expected in the best clinical practice. Future studies of dose-response relations could be enhanced by taking into account the principles noted above, and currently available data should be reanalyzed based on these principles. This re-analysis is performed in a companion article [Baker et al. 2003, Depress Anxiety 17:1-9]. PMID- 12577274 TI - Factorial validity of the center for epidemiologic studies-depression (CES-D) scale in military peacekeepers. AB - Despite widespread use of the Center for Epidemiologic Studies Depression Scale [CES-D], there are no investigations that examine its factor structure in a military sample. Separate confirmatory factor analyses were performed on responses to the CES-D obtained from 102 female and 102 male Canadian military peacekeepers in order to compare the fit of a four-factor intercorrelated (lower order) model to a four-factor hierarchical (higher-order) model. The intercorrelated and hierarchical models fit the data well for both women and men, with hierarchical models fitting the data slightly better for women than men. These findings suggest that, for military women and men, the CES-D can be used to measure a set of distinct but interrelated depressive symptoms as well as a global construct of depression. Implications and future directions are discussed. PMID- 12577275 TI - A model of risk for major depression: effects of life stress and cognitive style vary by age. AB - Empirical studies increasingly attribute risk for depression to adverse life events, cognitive style, and possibly to the interaction between cognitive style and event type. We present an evidence-based model, developed with independent samples of adults and elderly adults, indicating that risk for major depression associated with these factors varies with age. According to the model, adverse events and need for control, the cognitive style that is a key feature of Beck's concept of autonomy, are significant risk factors for depression in younger adults but not in elderly adults. The cognitive style of sociotropy, characterized by a high need for relatedness and concern about disapproval, is a stable risk factor, independent of age, in posing a risk for depression. The effects of the interactions of adverse event type (achievement events and interpersonal events) and cognitive style in predicting depression each appear to vary with age, expanding prior work, which suggests that adverse events affecting one's personal vulnerability are likely to precipitate depression. Age-specific approaches to reducing risk for major depression are clinically important, and the model presented here suggests that the use of an age-specific perspective would advance research in stress-diathesis models for risk of depression. PMID- 12577276 TI - Childhood adversities associated with major depression and/or anxiety disorders in a community sample of Ontario: issues of co-morbidity and specificity. AB - It has been well established that early adversity is a major risk factor for depression and for anxiety disorders in various populations and age groups. Few studies have considered the relative strength of these associations and the possible role of co-morbid depression/anxiety in understanding them. Using data from a large community sample of Ontario, Canada, we examined the relative strength of the associations between early physical abuse, sexual abuse, and/or parental strain with depression alone, anxiety alone, and co-morbid depression/anxiety. The current sample consisted of 6,597 individuals 15-64 years of age who were interviewed using the World Health Organization Composite International Diagnostic Interview (CIDI). Using a multivariate design, we compared early adversity scores across four diagnostic study groups including normal controls, individuals with major depression but no anxiety disorders, individuals with one or more anxiety disorders without major depression, and individuals with co-morbid major depression and anxiety. Individuals with past disorders were considered separately from those with current disorders. For both past and current disorders, highly significant differences in early adversity scores were found across the four study groups. A novel and robust finding, consistent across all analyses, was a marked association between early sexual abuse and co-morbid depression and anxiety but not the "pure" disorders. A strong association between early parental strain and major depression (independent of anxiety) was also found. The overall pattern of results suggest that there may be unique relationships linking particular adversities to particular manifestations of depression and anxiety disorders later in life. A particularly strong association between early sexual abuse and co-morbid depression/anxiety was found. PMID- 12577279 TI - Nanoelectrospray-based detection and sequencing of substoichiometric amounts of phosphopeptides in complex mixtures. AB - A novel immobilized metal affinity chromatography (IMAC)-based protocol was successfully used for sample preparation prior to nanoelectrospray-based sequencing of phosphopeptides. In a first step, phosphorylated peptides are detected in an unseparated peptide mixture using precursor ion scanning in the negative ion mode on a triple-quadrupole mass spectrometer. Then the phosphorylated peptides are enriched by passing the sample over an IMAC column and sequenced in the positive ion mode. The IMAC-based enrichment allows the sequencing of phosphorylated peptides even if other, much more abundant, peptides are present at the same m/z value in the original mixture. Using this two-step approach, we are able to combine the simplicity, sensitivity and specificity of precursor ion scanning-based detection of phosphopeptides using the nanoelectrospray ion source with the ability to sequence phosphorylated peptides even if they are present in substoichiometric amounts. PMID- 12577277 TI - Lithium augmentation increases post-dexamethasone cortisol in the dexamethasone suppression test in unipolar major depression. AB - Although considerable evidence exists on the efficacy of lithium as an augmenting agent in refractory depression, the underlying neurobiology of this phenomenon is unknown. In patients with major depression, changes of the hypothalamic-pituitary adrenocortical (HPA) system have been detected by means of the dexamethasone suppression test (DST), when administered during treatment with tricyclic antidepressants. We investigated whether the DST also reveals alterations of the HPA system during lithium augmentation. We also sought to identify whether response to lithium augmentation can be predicted with the DST. Twenty-five patients with unipolar major depression, who did not respond to an adequate antidepressant monotherapy of at least 4 weeks, were measured for basal (pre dexamethasone, 0800h) cortisol and ACTH levels and were administered the DST the day before initiation of lithium augmentation treatment. The same neuroendocrine procedures were repeated after 3 to 4 weeks. Criteria of response to lithium augmentation, defined as a reduction of the Hamilton Depression Rating Scale (HDRS17) score by > or =50% and an end point score of 9 or less, were determined by weekly HDRS ratings. The DST revealed a statistically significant increase of the post-dexamethasone cortisol values (P = 0.021) and an increase in the post dexamethasone ACTH values (P = 0.051) during lithium augmentation as compared to pre-treatment baseline evaluations. The pre-dexamethasone hormone values were unchanged. The number of non-suppressors at baseline was one and increased to three at follow-up. Results of DST did not predict response to lithium augmentation, which occurred in 40% of subjects. Results suggest that lithium augmentation increases HPA system activity, as indicated by the increase of post dexamethasone cortisol and ACTH levels measured by the DST. This is in contrast to the established decline of HPA system activity during treatment with tricyclic antidepressants. PMID- 12577280 TI - Rapid screening and characterization of drug metabolites using a new quadrupole linear ion trap mass spectrometer. AB - The application of a new hybrid RF/DC quadrupole-linear ion trap mass spectrometer to support drug metabolism and pharmacokinetic studies is described. The instrument is based on a quadrupole ion path and is capable of conventional tandem mass spectrometry (MS/MS) as well as several high-sensitivity ion trap MS scans using the final quadrupole as a linear ion trap. Several pharmaceutical compounds, including trocade, remikiren and tolcapone, were used to evaluate the capabilities of the system with positive and negative turbo ionspray, using either information-dependent data acquisition (IDA) or targeted analysis for the screening, identification and quantification of metabolites. Owing to the MS/MS in-space configuration, quadrupole-like CID spectra with ion trap sensitivity can be obtained without the classical low mass cutoff of 3D ion traps. The system also has MS(3) capability which allows fragmentation cascades to be followed. The combination of constant neutral loss or precursor ion scan with the enhanced product ion scan was found to be very selective for identifying metabolites at the picogram level in very complex matrices. Owing to the very high cycle time and, depending on the mass range, up to eight different MS experiments could be performed simultaneously without compromising chromatographic performance. Targeted product ion analysis was found to be complementary to IDA, in particular for very low concentrations. Comparable sensitivity was found in enhanced product ion scan and selected reaction monitoring modes. The instrument is particularly suitable for both qualitative and quantitative analysis. PMID- 12577281 TI - Electrospray mass spectra of group 6 (Fischer) carbenes in the presence of electron-donor compounds. AB - Fischer carbene complexes 1-7 are not ionized under standard electrospray ionization (ESI) conditions. We report here that unsaturated chromium and tungsten (Fischer) carbene complexes can be ionized in an electrospray ion source in the presence of electron-donor compounds such as hydroquinone (HQ) or tetrathiafulvalene (TTF). The addition of these compounds, which seem to act as electron transfer agents, permits the recording and study of their ESI mass spectra in the negative mode of detection. Both chromium and tungsten(0) carbene complexes undergo in the first fragmentation stage a double simultaneous decarbonylation process. PMID- 12577282 TI - Rapid quantification of HIV protease inhibitors in human plasma by high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. AB - HIV protease inhibitors are important antiretroviral drugs which have substantially reduced the morbidity and mortality associated with HIV-1 infection. Recent data have shown relationships between plasma concentrations of the protease inhibitors and clinical response, which makes therapeutic drug monitoring valuable. We have developed and validated an assay, using liquid chromatography coupled with electrospray tandem mass spectrometry (LC/MS/MS), for the routine quantification of the six licensed protease inhibitors (amprenavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir) and the pharmacologically active nelfinavir metabolite M8 in plasma. The sample pretreatment consisted of protein precipitation with a mixture of methanol and acetronitrile using only 100 microl of plasma. Chromatographic separation was performed on an Inertsil ODS3 column (50 x 2.0 mm i.d., particle size 5 microm), with a quick stepwise gradient using an acetate buffer (pH 5) and methanol, at a flow rate of 0.5 ml min(-1). The analytical run time was 5.5 min. The use of a 96 well plate autosampler allowed batch sizes up to 150 patient samples. The triple quadrupole mass spectrometer was operated in the positive ion mode and multiple reaction monitoring was used for drug quantification. The method was validated over the concentration ranges 0.01-10 microg ml(-1) for indinavir and saquinavir, 0.1-10 microg ml(-1) for amprenavir, 0.05-10 microg ml(-1) for nelfinavir and ritonavir, 0.1-20 microg ml(-1) for lopinavir and 0.01-5 microg ml(-1) for M8. Saquinavir-d(5) and indinavir-d(6) were used as internal standards. The coefficients of variation were always <10% for both intra-day and inter-day precisions for each compound. Mean accuracies were also between the designated limits (+/-15%). The validated concentration ranges proved to be adequate in daily practice. This robust and fast LC/MS/MS assay is now successfully applied for routine therapeutic drug monitoring and pharmacokinetic studies in our hospital. PMID- 12577283 TI - Mass spectrometry of polyaromatic sulfur compounds in the presence of palladium(II). AB - Electrospray ionization mass spectrometry (ESI-MS) and tandem MS in an ion trap were used to explore the reduction of a PdCl(2) solution and the role of the metal in the formation of organosulfur radical cations. For tandem MS experiments, a 32 Da neutral loss (M - S), was observed for dibenzothiophene, thianthrene and benzonaphthothiophene, whereas both 32 and 33 Da neutral losses were observed for 2-methyldibenzothiophene and 4,6-dimethyldibenzothiophene. The effects of several variables on the formation of molecular ions or molecular fragments were studied to optimize radical cation formation, including the concentration ratio between PdCl(2) and sulfur and the collision energy. Electrochemical and chromatographic data were used to provide an explanation for the relative sensitivity observed for a mixture of polyaromatic sulfur heterocycles. PMID- 12577284 TI - Fragmentation reactions of protonated peptides containing glutamine or glutamic acid. AB - A variety of protonated dipeptides and tripeptides containing glutamic acid or glutamine were prepared by electrospray ionization or by fast atom bombardment ionization and their fragmentation pathways elucidated using metastable ion studies, energy-resolved mass spectrometry and triple-stage mass spectrometry (MS(3)) experiments. Additional mechanistic information was obtained by exchanging the labile hydrogens for deuterium. Protonated H-Gln-Gly-OH fragments by loss of NH(3) and loss of H(2)O in metastable ion fragmentation; under collision-induced dissociation (CID) conditions loss of H-Gly-OH + CO from the [MH - NH(3)](+) ion forms the base peak C(4)H(6)NO(+) (m/z 84). Protonated dipeptides with an alpha-linkage, H-Glu-Xxx-OH, are characterized by elimination of H(2)O and by elimination of H-Xxx-OH plus CO to form the glutamic acid immonium ion of m/z 102. By contrast, protonated dipeptides with a gamma-linkage, H-Glu(Xxx-OH)-OH, do not show elimination of H(2)O or formation of m/z 102 but rather show elimination of NH(3), particularly in metastable ion fragmentation, and elimination of H-Xxx-OH to form m/z 130. Both the alpha- and gamma-dipeptides show formation of [H-Xxx-OH]H(+), with this reaction channel increasing in importance as the proton affinity (PA) of H-Xxx-OH increases. The characteristic loss of H(2)O and formation of m/z 102 are observed for the protonated alpha tripeptide H-Glu-Gly-Phe-OH whereas the protonated gamma-tripeptide H-Glu(Gly-Gly OH)-OH shows loss of NH(3) and formation of m/z 130 as observed for dipeptides with the gamma-linkage. Both tripeptides show abundant formation of the y(2)'' ion under CID conditions, presumably because a stable anhydride neutral structure can be formed. Under metastable ion conditions protonated dipeptides of structure H-Xxx-Glu-OH show abundant elimination of H(2)O whereas those of structure H-Xxx Gln-OH show abundant elimination of NH(3). The importance of these reaction channels is much reduced under CID conditions, the major fragmentation mode being cleavage of the amide bond to form either the a(1) ion or the y(1)'' ion. Particularly when Xxx = Gly, under CID conditions the initial loss of NH(3) from the glutamine containing dipeptide is followed by elimination of a second NH(3) while the initial loss of H(2)O from the glutamic acid dipeptide is followed by elimination of NH(3). Isotopic labelling shows that predominantly labile hydrogens are lost in both steps. Although both [H-Gly-Glu-Gly-OH]H(+) and [H-Gly Gln-Gly-OH]H(+) fragment mainly to form b(2) and a(2) ions, the latter also shows elimination of NH(3) plus a glycine residue and formation of protonated glycinamide. Isotopic labelling shows extensive mixing of labile and carbon bonded hydrogens in the formation of protonated glycinamide. PMID- 12577285 TI - Differentiation of a pair of diastereomeric tertiarybutoxycarbonylprolylproline ethyl esters by collision-induced dissociation of sodium adduct ions in electrospray ionization mass spectrometry and evidence for chiral recognition by ab initio molecular orbital calculations. AB - The fragmentation of the sodium adduct ions for tert-butoxycarbonyl-L-prolyl-L proline ethyl ester (Boc-L-Pro-L-Pro-OEt) was compared with that for Boc-D-Pro-L Pro-OEt in positive-ion electrospray ionization (ESI) mass spectrometry. In the collision-induced dissociation (CID) mass spectra of the [M + Na](+) ions, the abundance of the [M + Na - C(CH(3))(3) + H](+) ion, which is due to the loss of a tert-butyl group from the [M + Na](+) ion for Boc-D-Pro-L-Pro-OEt, was about eight times higher than that for Boc-L-Pro-L-Pro-OEt. In addition, in the CID spectra of the sodium adduct fragment ion ([M + Na - Boc + H](+)), the abundance of the [M + Na - Boc - prolylresidue + H](+) ion, which is due to the loss of prolyl residue from the [M + Na - Boc + H](+) ion for Boc-L-Pro-L-Pro-OEt, was about five times higher than that for Boc-D-Pro-L-Pro-OEt. These results indicate that Boc-L-Pro-L-Pro-OEt was distinguished from Boc-D-Pro-L-Pro-OEt by the CID mass spectra of the sodium adduct ions in ESI mass spectrometry. The optimized geometries of the [M + Na](+) and the [M + Na - Boc + H](+) ions calculated by ab initio molecular orbital calculations suggest that the chiral recognition of these diastereomers was due to the difference of the orientation of a sodium ion to the oxygen and nitrogen atoms in dipeptide derivatives, and to the difference of the total energies between them. PMID- 12577286 TI - Accurate mass measurements by Fourier transform mass spectrometry in the study of advanced glycation end products/peptides. AB - The Maillard reaction occurring between sugars and amino groups is important in living systems. When amino groups belonging to protein chains are involved, the Maillard reaction has been invoked as responsible for protein cross-linking and the production of 'toxic' compounds. The reaction leads to the production of a heterogeneous group of substances, usually called advanced glycation end products (AGEs). Classical analytical approaches, such as spectroscopic (ultraviolet, fluorescence) and mass spectrometric (matrix-assisted laser desorption/ionization, liquid chromatography/electrospray ionization mass spectrometry) methods, have shown that the digestion mixture is highly complex. However, there are clear differences between the digestion mixtures of glycated and unglycated human serum albumin (HSA). In the former case, possible glycated peptides belonging to the AGE peptide class may be identified. Tandem mass spectrometric experiments on selected species seemed to be promising as regards structural information, but it was thought of interest to undertake the present investigation, based on liquid chromatography/electrospray ionization Fourier transform mass spectrometry, in order to obtain definitive results on their elemental composition. Using this approach, about 20 glycated peptides were detected and their possible structures were postulated by examining the known sequence of HSA. PMID- 12577287 TI - Deconvolution of mass spectra measured with a non-uniform detector array to give accurate ion abundances. AB - Array detectors are under active development since they offer large improvements in the efficiency of mass spectrum measurements. High quality is always a requirement whether array detectors are used for ions, electrons, UV, x-rays, etc., but in mass spectrometry the very high accuracy frequently needed in ion abundance measurements for isotope ratios is unique. These demands necessitate modelling the measurement process and careful deconvolution of the measured data. A linear model in terms of matrix algebra is presented in which the incident spectrum (unknown) and the measured spectrum are represented by column matrices and the action of the detector array on the incident spectrum is represented by an experimentally measurable square 'A' matrix. Residual noise in this matrix can be minimized following a singular value decomposition procedure and its use in 'forward deconvolution' of measured spectra is discussed. The random error in the incident ion counts is accounted for after the deconvolution since this is not an error from the perspective of the detector. The microchannel plate electron multiplier gain distribution is an important feature of the deconvolution. PMID- 12577288 TI - Addressing the metabolic activation potential of new leads in drug discovery: a case study using ion trap mass spectrometry and tritium labeling techniques. AB - Metabolic activation of drug candidates to electrophilic reactive metabolites that can covalently modify cellular macromolecules may result in acute and/or idiosyncratic immune system-mediated toxicities in humans. This presents a significant potential liability for the future development of these compounds as safe therapeutic agents. We present here an example of an approach where sites of metabolic activation within a new drug candidate series were rapidly identified using online liquid chromatography/multi-stage mass spectrometry on an ion trap mass spectrometer. This was accomplished by trapping the reactive intermediates formed upon incubation of compounds with rat and human liver microsomes as their corresponding glutathione conjugates and mass spectral characterization of these thiol adducts. Based on the structures of the GSH adducts identified, potential sites and mechanisms of bioactivation within the chemical structure were proposed. These metabolism studies were interfaced with iterative structural modifications of the chemical series in order to block these bioactivation sites within the molecule. This strategy led to a significant reduction in the propensity of the compounds to undergo metabolic activation as evidenced by reductions in the irreversible binding of radioactivity to liver microsomal material upon incubation of tritium-labeled compounds with this in vitro system. With the efficiency and throughput achievable with such an approach, it appears feasible to identify and address the metabolic activation potential of new drug leads during routine metabolite identification studies in an early drug discovery setting. PMID- 12577289 TI - Non-linearity in the quadrupole detector system: implications for the determination of the 13C mass distribution of an ion fragment. AB - Some biomedical (13)C tracer applications to study glucose/lactate metabolism require the determination of up to 7 mass isotopomers of glucose, generated by the (13)C-labeling. Isotopomer ratios determined by GC-Quadrupole-MS can depend on the analyte amount being measured and may differ from expected theoretical values. A calibration is necessary to correct these effects. However, the large number of different isotopomers prevents an empirical calibration, that is based on known isotopomer mixtures. Neither insufficient mass separation in the quadrupole nor proton adducts or losses of the ionized fragment can account for the observed deviation from expected values and the dependency on the sample size. We explain the deviations with a nonlinear mapping of the isotopomer amount being analyzed to the area under the elution curve for the corresponding mass channel. The nonlinear relation allows to estimate potential errors in the determination of isotopomer ratios. It also allows to define a dynamic range as the range of signal areas, for which the systematic error in the isotopomer ratio determination is below 5% of the nominal value. It is 1 : 200 and 1 : 500 for two different fragments of glucose, measured under EI and CI and is smaller than the range imposed by the measurement precision. After correcting the nonlinearities the dynamic range improved by the factor 10 in both cases. PMID- 12577291 TI - Current literature in journal of mass spectrometry. PMID- 12577290 TI - Interesting differences between the protonated and sodium adducts of pentacoordinated bisaminoacylspirophosphoranes in electrospray ionization mass spectrometry. PMID- 12577292 TI - Visualizing metabolic changes in breast-cancer tissue using 1H-NMR spectroscopy and self-organizing maps. AB - In-vitro NMR spectroscopic examinations of tissue extracts can be combined with appropriate pattern-recognition and visualization techniques in order to monitor characteristic metabolic differences between tissue classes. In the present study, such techniques are applied to a set of 88 breast-tissue samples with the intention of identifying typical differences between various tissue classes. The set contains 49 breast-tumor samples of various tumor grades and 39 samples of healthy tissue. The metabolite compositions of the tissue extracts were investigated using a dual extraction technique and high-resolution (1)H-NMR spectroscopy. The spectra of the hydrophilic and the lipophilic compounds were assigned to three groups according to different malignancy grades of the respective tissue samples. The group characteristics were analyzed using the k nearest-neighbor method and self-organizing-map visualizations. The results show an increase of UDP-hexose, phosphocholine and phosphoethanolamine concentrations according to the tumor grade. Higher concentrations of taurine were detected in the malignant samples. Myo-inositol and glucose content were elevated in control samples compared with malignant tissue. Both compounds also characterized different subgroups in the pool of unaffected tissue samples depending upon fat content or fibrosis. Several lipid metabolites showed a characteristic elevation with high malignancy. PMID- 12577293 TI - Characterization of oligodendrogliomas using short echo time 1H MR spectroscopic imaging. AB - Oligodendroglial tumors may not be distinguished easily from other brain tumors based on clinical presentation and magnetic resonance imaging (MRI) alone. Identification of these tumors however may have therapeutic consequences. The purpose of this study was to characterize and identify oligodendrogliomas by their metabolic profile as measured by (1)H MR spectroscopic imaging (MRSI). Fifteen patients with oligodendroglial tumors (eight high-grade oligodendrogliomas, seven low-grade oligodendrogliomas) underwent MRI and short echo time (1)H MRSI examinations. Five main metabolites found in brain MR spectra were quantified and expressed as ratios of tumor to contralateral white matter tissue. The level of lipids plus lactate was also assessed in the tumor. For comparison six patients with a low grade astrocytoma were also included in the study. The metabolic profile of oligodendrogliomas showed a decreased level of N acetylaspartate and increased levels of choline-containing compounds and glutamine plus glutamate compared with white matter. The level of glutamine plus glutamate was significantly higher in low-grade oligodendrogliomas than in low grade astrocytomas and may serve as a metabolic marker in diagnosis and treatment planning. In high-grade oligodendrogliomas large resonances of lipids plus lactate were observed in contrast to low-grade tumors. PMID- 12577294 TI - Saturation transfer in human red blood cells with normal and unstable hemoglobin. AB - Saturation transfer phenomena from irradiated protein protons to observed water protons in packed human red blood cells (RBCs) with normal or unstable hemoglobin (Hb), i.e. Hb Yokohama and Hb Koeln, were studied using intermolecular cross relaxation rates [CR; 1/T(IS)(H(2)O)], action spectra [[1-(I(infinity)/I(0))] vs f(2) (ppm), where I(0) and I(infinity) are the longitudinal magnetization of observed water protons before and after long-time f(2)-irradiation, respectively], CR spectra [CR vs f(2) (ppm)] and CR ratio vs f(2) (ppm) with f(2) irradiation from -100 to 100 ppm at gammaH(2)/2pi of 69 or 250 Hz. RBCs (Hb Yokohama) exhibited many large Heinz bodies and strongly impaired filterability, while RBCs (Hb Koeln) showed few microscopically typical Heinz bodies and virtually normal filterability. However, increases in CR values for RBCs (Hb Koeln) and RBCs (Hb Yokohama), monitored by f(2)-irradiation below approximately 6 and above approximately 14 ppm, clearly indicated marked increases in association or aggregation of unstable Hb in RBCs compared with those in normal RBCs. CR values, monitored between approximately 0 and approximately 10 ppm, were related to not only association or aggregation of unstable Hb but also amounts of water in RBCs. Aggregation or association of unstable Hb exhibited greater effects on CR values compared with those of methemoglobin formation. PMID- 12577295 TI - Adiabatic RARE imaging. AB - The practical implementation of ultra-fast spin-echo, or RARE imaging with adiabatic RF pulses and surface coil transmission at 7 T is described. Despite the large RF inhomogeneities, the adiabatic character of the 180 degrees BIR-4 refocusing pulses ensures optimal sensitivity and minimal image artifacts. An internal 'phase-cycle' is used to remove spurious unwanted coherences. The short T(2) relaxation times in rat brain at 7 T demand a centric, rather than a linear coverage of k-space in order to avoid excessive signal loss. T(2) relaxation during k-space coverage also leads to image blurring, which can be counteracted by interleaved k-space sampling. The coverage of k-space in four acquisitions provides high-quality anatomical images comparable to conventional spin-echo images. A two-scan RARE implementation provides sufficient spatial and temporal resolution for most applications. Quantitative mapping of T(1) relaxation and cerebral blood flow changes during forepaw stimulation in the rat are demonstrated. PMID- 12577296 TI - Quantification of the glycogen 13C-1 NMR signal during glycogen synthesis in perfused rat liver. AB - We studied glycogen synthesis from glucose in perfused livers of fed (n = 4) and 24 h starved (n = 7) rats. Glycogenolysis was inhibited by BAY R3401 (150 microM) and proglycosyn (100 microM). After 60 min, we replaced 99% (13)C-1 glucose by natural abundance glucose. This pulse-chase design allowed us to recognize residual ongoing futile glycogen turnover from the release of initially deposited (13)C-label, into the (13)C-free chase medium. Net residual turnover was less than 2 +/- 0.7% and 0.6 +/- 0.2% of 1-(13)C glycogen deposition rates of 0.31 +/- 0.04 and 0.99 +/- 0.04 micromol glucose g(-1) min(-1), in starved and fed livers, respectively. The 1-(13)C glycogen signal was monitored throughout the experiment with proton-decoupled (13)C NMR spectroscopy and analyzed in the time domain using AMARES. We noticed progressive line-broadening in any single experiment in the chase phase. One or a sum of two to three overlapping Lorentzians, with different exponential damping factors, were fitted to the signal. When the S/N was better than 40, the fit always delivered a small and a broad component. In the chase phase, the fit with a single Lorentzian resulted in a decline of glycogen signal by about 15 +/- 4 and 12 +/- 2% in starved and fed rats, respectively. This apparent decline in 1-(13)C glycogen signal could not be accounted for by the appearance of equivalent amounts of (13)C-labeled metabolites in the perfusate. The fit with a sum of two Lorentzians resulted in a decline of glycogen signal intensity of 7 +/- 5 and 5 +/- 3% in starved and fed rats, respectively, which reduced the apparent turnover to 8 +/- 9% and 6 +/- 4%, respectively. Quantification of the growing (13)C-1 glycogen signal requires a model function that accommodates changes in line shape throughout the period under study. PMID- 12577297 TI - In vivo monitoring of rat brain metabolites during vigabatrin treatment using localized 2D-COSY. AB - A two-dimensional COSY-based localization sequence was designed to allow the in vivo monitoring of proton metabolites in rat brain [particularly gamma aminobutyric acid (GABA), glutamine, taurine and myo-inositol]. The sequence incorporated OSIRIS signal localization, B1-insensitive water suppression and phase-sensitive COSY acquisition. The method was used to study the effects of the GABA-transaminase inhibitor vigabatrin on rat brain metabolite concentrations. Wistar rats were treated daily for 3 days with an oral dose of vigabatrin (200 mg/kg, n = 4). Localized COSY spectra were obtained during a 120 min acquisition from a 270 microl central brain voxel and compared with nine untreated control animals. Significant elevations were observed in GABA (267% of control, p < 0.005, Mann-Witney test), glutamine (130% of control, p < 0.005) and taurine (113% of control, p < 0.05). Changes in GABA and taurine were consistent with previous data on the action of Vigabatrin, and support a previously hypothesized link between these compounds. The increase in glutamine was more surprising and may reflect the balance between the level and/or site of GABA-transaminase inhibition and downregulation of GABA synthesis. PMID- 12577298 TI - Conservative segregation of maternally inherited CS histone variants in larval stages of sea urchin development. AB - Three sets of histone variants are coexisting in the embryo at larval stages of sea urchin's development: the maternally inherited cleavage stage variants (CS) expressed during the two initial cleavage divisions, the early histone variants, which are recruited into embryonic chromatin from middle cleavage stages until hatching and the late variants, that are fundamentally expressed from blastula stage onward. Since the expression of the CS histones is confined to the initial cleavage stages, these variants represent a very minor proportion of the histones present in the plutei larvae, whereas the late histone variants are predominant. To determine the position of these CS in the embryonic territories, we have immunolocalized the CS histone variants in plutei larvas harvested 72 h post fertilization. In parallel, we have pulse labeled the DNA replicated during the initial cleavage cycle with bromodeoxyuridine (BrdU) and its position was further determined in the plutei larvas by immunofluorescence. We have found that the CS histone variants were segregated to specific territories in the plutei. The position in which the CS histone variants were found to be segregated was consistent with the position in which the DNA molecules that were replicated during the initial cleavage divisions were localized. These results strongly suggest that a specification of embryonic nuclei occurs at the initial cleavage divisions which is determined by a chromatin organized by CS histone variants. PMID- 12577299 TI - Inflammatory T cells rapidly induce differentiation of human bone marrow stromal cells into mature osteoblasts. AB - Activated T cells secrete multiple osteoclastogenic cytokines which play a major role in the bone destruction associated with rheumatoid arthritis. While the role of T cells in osteoclastogenesis has received much attention recently, the effect of T cells on osteoblast formation and activity is poorly defined. In this study, we investigated the hypothesis that in chronic inflammation activated T cells contribute to enhanced bone turnover by promoting osteoblastic differentiation. We show that T cells produce soluble factors that induce alkaline phosphatase activity in bone marrow stromal cells and elevated expression of mRNA for Runx2 and osteocalcin. This data indicate that T cell derived factors have the capacity to stimulate the differentiation of bone marrow stromal cells into the osteoblast phenotype. RANKL mRNA was undetectable under any conditions in highly purified bone marrow stromal cells. In contrast, RANKL was constitutively expressed in primary osteoblasts and only moderately up-regulated by activated T cell conditioned medium. Interestingly, both bone marrow stromal cells and osteoblasts expressed mRNA for RANK, which was strongly up-regulated in both cell types by activated T cell conditioned medium. Although, mRNA for the RANKL decoy receptor, osteoprotegerin, was also up-regulated by activated T cell conditioned medium, it's inhibitory effects may be mitigated by a simultaneous rise in the osteoprotegerin competitor TNF-related apoptosis-inducing ligand. Based on our data we propose that during chronic inflammation, T cells regulate bone loss by a dual mechanism involving both direct stimulation of osteoclastogenesis, by production of osteoclastogenic cytokines, and indirectly by induction of osteoblast differentiation and up-regulation of bone turnover via coupling. PMID- 12577301 TI - G1 tetraploidy checkpoint and the suppression of tumorigenesis. AB - Checkpoints suppress improper cell cycle progression to ensure that cells maintain the integrity of their genome. During mitosis, a metaphase checkpoint requires the integration of all chromosomes into a metaphase array in the mitotic spindle prior to mitotic exit. Still, mitotic errors occur in mammalian cells with a relatively high frequency. Metaphase represents the last point of control in mitosis. Once the cell commits to anaphase there are no checkpoints to sense segregation defects. In this context, we will explore our recent finding that non transformed mammalian cells have a checkpoint that acts subsequent to mitotic errors to block the proliferation of cells that have entered G1 with tetraploid status. This arrest is dependent upon both p53 and pRb, and may represent an important function of both p53 and pRb as tumor suppressors. Further, we discuss the possibility that this mechanism may similarly impose G1 arrest in cells that become aneuploid through errors in mitosis. PMID- 12577300 TI - Lysyl oxidase: properties, specificity, and biological roles inside and outside of the cell. AB - Lysyl oxidase (LO) plays a critical role in the formation and repair of the extracellular matrix (ECM) by oxidizing lysine residues in elastin and collagen, thereby initiating the formation of covalent crosslinkages which stabilize these fibrous proteins. Its catalytic activity depends upon both its copper cofactor and a unique carbonyl cofactor and has been shown to extend to a variety of basic globular proteins, including histone H1. Although the three-dimensional structure of LO has yet to be determined, the present treatise offers hypotheses based upon its primary sequence, which may underlie the prominent electrostatic component of its unusual substrate specificity as well as the catalysis-suppressing function of the propeptide domain of prolysyl oxidase. Recent studies have demonstrated that LO appears to function within the cell in a manner, which strongly modifies cellular activity. Newly discovered LO-like proteins also likely play unique roles in biology. PMID- 12577302 TI - Chromatin remodeling as a guide to transcriptional regulatory networks in mammals. AB - An important challenge of genome biology is a dissection of transcriptional regulatory networks that operate inside the nucleus during ontogeny and disease (Wyrick and Young [2002] Curr. Opin. Genet. Dev. 12:130). Limitations of existing experimental tools greatly complicate such analysis in the human genome: for example, genome-wide expression profiling of cells responding to a stimulus fails to reveal a majority of the genes involved in the functional network of responding to that stimulus [Giaver et al., 2002; Birrell et al., 2002]. This article discusses recent advances in analyzing mammalian transcriptional regulatory circuits [Nikiforov et al., 2002; Weinmann et al., 2002; Ren et al., 2000]. As evidenced by these and other data, paucity of information about the location of regulatory DNA elements in the human genome presents an obstacle to comprehensive transcription network analysis. It has been known since the late 1970s that chromatin over active regulatory DNA stretches is stably remodeled into "nuclease hypersensitive sites" [Elgin, 1988; Gross and Garrard 1988]. Massively parallel analysis of such remodeling in cell nuclei identifies regulatory DNA that is difficult to map comprehensively using other approaches, reveals genes poised for rapid activation, and offers a novel perspective on the "epigenome"--the regulatory program being executed by the genome in a given cell type. PMID- 12577303 TI - New insights into the mechanisms of vitamin D action. AB - The biologically active metabolite of vitamin D, 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) is a secosteroid whose genomic mechanism of action is similar to that of other steroid hormones and is mediated by stereospecific interaction of 1,25(OH)(2)D(3) with the vitamin D receptor (VDR) which heterodimerizes with the retinoid X receptor (RXR). After interaction with the vitamin D response element (VDRE) in the promoter of target genes, transcription proceeds through the interaction of VDR with coactivators and with the transcription machinery. The identification of the steps involved in this process has been a major focus of recent research in the field. However, the functional significance of target proteins as well as the functional significance of proteins involved in the transport and metabolism of vitamin D is also of major importance. Within the past few years much new information has been obtained from studies using knockout and transgenic mice. New insight has been obtained using this technology related to the physiological significance of the vitamin D binding protein (DBP), used to transport vitamin D metabolites, as well as the physiological significance of target proteins including 25-hydroxyvitamin D(3) 24-hydroxylase (24(OH)ase), 25 hydroxyvitamin D(3)-1 alpha-hydroxylase (1 alpha-(OH)ase), VDR, and osteopontin. The crystal structure of the DBP and the ligand binding domain of the VDR have recently been reported, explaining, in part, the unique properties of these proteins. In addition novel 1,25(OH)(2)D(3) target genes have been identified including the epithelial calcium channel, present in the proximal intestine and in the distal nephron. Thus in recent years a number of exciting discoveries have been made that have enhanced our understanding of mechanisms involved in the pleiotropic actions of 1,25(OH)(2)D(3). PMID- 12577304 TI - A unified model for the action of leptin on bone turnover. AB - Leptin has been advocated as a centrally acting factor responsible for inhibiting accumulation of bone mass. However, recent investigations unequivocally establish leptin as a local (autocrine) factor expressed by osteoblasts. Exogenously added leptin causes osteoblastic cell proliferation and differentiation, while also rendering osteoblasts more efficacious in terms of mineralization. Leptin acts as an anti-apoptotic agent, and augments messages responsible for the remodelling of bone tissue, i.e., mRNAs for osteoprotegerin (OPG) and the interleukin IL-6. Furthermore, leptin message is readily expressed in osteoblasts subjected to mechanical strain. In this respect, osteoblasts, which are unilaterally stretched proliferate and differentiate, a phenomenon being potentiated by exposure of the cells to differentiating humoral factors. This article discusses a unified model of dually acting leptin through the central nervous system and the mechanostat principle applied to osteoblasts. The proposed model may account for the finely tuned bone homeostasis maintained within rather narrow limits, depending on exposure to humoral factors and the prevailing mechanostat usage mode. PMID- 12577305 TI - Chondrogenesis induced by actin cytoskeleton disruption is regulated via protein kinase C-dependent p38 mitogen-activated protein kinase signaling. AB - Disruption of the actin cytoskeleton in subconfluent mesenchymal cells induces chondrogenic differentiation via protein kinase C (PKC) alpha signaling. In this study, we investigated the role of p38 mitogen-activated protein (MAP) kinase in the chondrogenic differentiation of mesenchymal cells that is induced by depolymerization of the actin cytoskeleton. Treatment of mesenchymal cells derived from chick embryonic limb buds with cytochalasin D (CD) disrupted the actin cytoskeleton with concomitant chondrogenic differentiation. The chondrogenesis was accompanied by an increase in p38 MAP kinase activity and inhibition of p38 MAP kinase with SB203580 blocked chondrogenesis. Together these results suggest an essential role for p38 MAP kinase in chondrogenesis. In addition, inhibition of p38 MAP kinase did not alter CD-induced increased expression and activity of PKC alpha, whereas down-regulation of PKC by prolonged exposure of cells to phorbol ester inhibited CD-induced p38 MAP kinase activation. Our results therefore suggest that PKC is involved in the regulation of chondrogenesis induced by disruption of the actin cytoskeleton via a p38 MAP kinase signaling pathway. PMID- 12577306 TI - Insulin-like and non-insulin-like selenium actions in 3T3-L1 adipocytes. AB - In insulin-sensitive 3T3-L1 adipocytes, selenium stimulates glucose transport and antilipolysis and these actions of selenium, like insulin actions, are sensitive to wortmanin, an inhibitor of phosphatidylinositol-3-kinase (PI3K). Selenium stimulates PI3K activity that is sustained up to 24 h. Selenium after 5-10 min increases tyrosine phosphorylation of selective cellular proteins, but after 24 h overall tyrosine phosphorylation is increased. Tyrosine phosphorylation of insulin receptor substrate 1 is detected when enriched by immunoprecipitation with anti-PI3K antibody. Selenium, however, does not stimulate insulin receptor tyrosine kinase activity. Selenium also increases phosphorylation of other insulin signaling proteins, including Akt and extracellular signal regulated kinases. Selenium-stimulated glucose transport is accompanied by increases in glucose transporter-1 content in the plasma membrane. These data are consistent with similar selenium action in glucose transport in 3T3-L1 fibroblasts expressing mainly GLUT1. In chronic insulin-induced insulin resistant cells, selenium unlike insulin fully stimulates glucose transport. In summary, selenium stimulates glucose transport and antilipolysis in a PI3K-dependent manner, but independent of insulin receptor activation. Selenium exerts both insulin-like and non-insulin-like actions in cells. PMID- 12577307 TI - Biochemical regulation of breast cancer cell expression of S1P2 (Edg-5) and S1P3 (Edg-3) G protein-coupled receptors for sphingosine 1-phosphate. AB - G protein-coupled receptors (GPCRs) for lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) transduce signals to many functions of normal cells. Most human cancer cells upregulate S1P and LPA GPCRs, in patterns distinctive for each type of tumor. The findings that 1-alpha, 25-dihydroxy vitamin D(3) (VD3) and all-trans retinoic acid (RA) differentially alter expression of the predominant S1P(3) (Edg-3) R and S1P(2) (Edg-5) R in human breast cancer cells (BCCs) permitted analyses of their individual activities, despite a lack of selective pharmacological probes. S1P-evoked increases in [Ca(2+)](i) in S1P(3) R-predominant BCCs were suppressed by concentrations of VD3 and RA which decreased expression of S1P(3) Rs, despite RA-induced increases in S1P(2) Rs. S1P-elicited chemokinetic migration of S1P(3) R-predominant BCCs across a type IV collagen-coated micropore filter also was inhibited by concentrations of VD3 and RA which decreased expression of S1P(3) Rs. The RA induced increase in expression of S1P(2) Rs did not prevent suppression by RA of S1P-elicited chemokinesis, which appears to be mediated by S1P(3) Rs, but instead exposed S1P(2) R-mediated inhibition of epidermal growth factor-stimulated chemotaxis of BCCs. In contrast, expression of the predominant LPA(2) Rs, LPA evoked increase in [Ca(2+)](i) and LPA-stimulated chemokinetic migration were suppressed concomitantly by RA but not VD3. Thus two structurally-homologous S1P Rs of BCCs differ in coupling to [Ca(2+)](i) signaling and have opposite effects on protein growth factor-stimulated chemotaxis. PMID- 12577308 TI - Characterization of the neuropilin-1 promoter; gene expression is mediated by the transcription factor Sp1. AB - Neuropilin-1 (NRP1) is a receptor for the vascular endothelial growth factor (VEGF) family of angiogenesis factors and for the semaphorin family of secreted neuronal guidance polypeptides. Very little is known, however, about how NRP1 gene expression is regulated. In this study, it was demonstrated that the tumor promoter, TPA (12-O-tetradecanoylphorbol-13-acetate) significantly up-regulated NRP1 mRNA levels by increasing its gene transcription rate in a manner dependent on de novo protein synthesis. To determine which elements regulate functional NRP1 expression, the promoter regions of human and mouse NRP1 genes were cloned and characterized. Promoter-reporter gene transfection experiments using deletion and point mutations demonstrated that two Sp1 elements are major contributors to both the constitutive and TPA-induced activity of the NRP1 promoter. Gel shift analysis showed a specific binding of the Sp1 transcription factor to those elements. Further mutational analysis revealed that an AP-1, and a CCAAT box also contributed to NRP1 constitutive and TPA-induced promoter activity. It was concluded that NRP1 expression is regulated by the cooperation of several regulatory elements including AP-1, Sp1, and a CCAAT box. PMID- 12577309 TI - Up-regulation of cDK5/p35 by oxidative stress in human neuroblastoma IMR-32 cells. AB - Cdk5, a member of the cyclin-dependent kinase (cdk) family, is predominantly active in neurons, where its activity is tightly regulated by the binding of its neuronal activators p35 and p39. Cdk5 is implicated in regulating the proper neuronal function; a deregulation of cdk5 has been found associated with Alzheimer's disease and amyotrophic lateral sclerosis. As oxidative stress products have been seen co-localized with pathological hallmarks of neurodegenerative diseases, we studied the effect of oxidative stress on the cdk5 enzyme in human neuroblastoma IMR-32 cells. We evaluated the effects of 4 hydroxynonenal and Ascorbate plus FeSO(4) on cdk5 activity and on the expression of cdk5 and p35 proteins. We report here that oxidative stress stimulates cdk5 activity and induces an upregulation of its regulatory and catalytic subunit expression in IMR-32 vital cells, showing that the cdk5 enzyme is involved in the signaling pathway activated by oxidative stress. PMID- 12577310 TI - Role of testicular interstitial macrophages in regulating testosterone release in hyperprolactinemia. AB - Hyperprolactinemia-induced hypogonadism has been linked to a dysfunction of the hypothalamus-pituitary-testis axis. The direct inhibitory effects of prolactin on the testicular release of testosterone have also been demonstrated, though their mechanisms remain unclear. Incubation of rat testicular interstitial cells (TICs) with prolactin stimulated the release of testosterone. TICs from rats with anterior pituitary-grafting-induced hyperprolactinemia release lower amounts of testosterone than controls. However, Leydig cells isolated from anterior pituitary-grafted rats release a greater amount of testosterone. These paradoxical observations have remained unexplained. This study examined the roles of testicular interstitial macrophages and of their product, tumor necrosis factor-alpha (TNF-alpha), in regulating Leydig cells under condition of hyperprolactinemia. Hyperprolactinemia was induced by grafting two anterior pituitary glands of rats under the renal capsule. Control animals were grafted with rat cortex tissue. The rats were sacrificed 6 weeks later. TICs and macrophages, and Leydig cells were isolated for in vitro incubation and drugs challenge. Testosterone released by testicular interstitial or Leydig cells was measured by radioimmunoassay. TNF-alpha concentration in the medium of TICs or macrophages was measured by enzyme-linked immunosorbent assay (ELISA). A dose dependent stimulation of TNF-alpha secretion in the medium of TICs or macrophages by the prolactin challenge was observed. Higher amounts of TNF-alpha were released by TICs in the anterior pituitary-grafted rats than in the control group. In contrast, the release of TNF-alpha by testicular interstitial macrophages isolated from the anterior pituitary- and cortex-grafted groups was quantitatively similar. Challenge with human chorionic gonadotropin did not modify the TNF-alpha release by testicular interstitial macrophages in either group. Challenge of Leydig cells with TNF-alpha inhibited their release of testosterone stimulated by human chorionic gonadotropin, but not their basal testosterone release. These different patterns of testosterone release in TICs versus Leydig cells cultures in anterior pituitary-grafted rats may be due to the influence of testicular interstitial macrophages. These observations correlate with in vivo conditions, where prolactin increases the release of TNF-alpha by testicular interstitial macrophages, which, in turn, decreases the human chorionic gonadotropin-stimulated release of testosterone by Leydig cells. In summary, hyperprolactinemia-induced hypogonadism involves a mechanism of prolactin-originated, macrophage-mediated inhibitory regulation of testosterone release by Leydig cells. TNF-alpha, one of the cytokines secreted by macrophages, may play a key role in this mechanism. PMID- 12577311 TI - TGF-beta-induced protein beta ig-h3 is upregulated by high glucose in vascular smooth muscle cells. AB - TGF-beta-induced gene-h3 (beta ig-h3) is an adhesive molecule that interacts with integrins. Because TGF-beta plays an important role in diabetic complications and beta ig-h3 serves as a cell substrate, we hypothesized that diabetic conditions might increase beta ig-h3 synthesis in vascular smooth muscle cells (VSMCs), which may subsequently contribute to the pathogenesis of diabetic angiopathy. The concentrations of beta ig-h3 and TGF-beta were measured in conditioned media using an enzyme-linked immunosorbent assay. An immunohistochemical study showed that beta ig-h3 was expressed in the VSMCs and the matrix of rat aortas. TGF-beta stimulated beta ig-h3 production, and high glucose induced beta ig-h3 as well as TGF-beta production in the VSMCs. The high glucose-induced beta ig-h3 expression was almost entirely blocked by an anti-TGF-beta antibody. beta ig-h3 protein mediated the adhesion, spreading, migration, and proliferation of rat VSMCs. These results suggest that the high glucose-induced beta ig-h3 in VSMCs regulates VSMC functions and may play an important role in diabetic angiopathy. PMID- 12577312 TI - Differential roles of hydrogen peroxide and superoxide in mediating IL-1-induced NF-kappa B activation and iNOS expression in bovine articular chondrocytes. AB - Our previous studies showed that reactive oxygen species (ROS) are required for the pro-inflammatory cytokine interleukin-1 beta (IL-1) to induce the activity of the Nuclear transcription Factor-kappa B (NF-kappa B) and the expression of the inducible isoform of the nitric oxide synthase (iNOS) in bovine articular chondrocytes. This study aimed at elucidating the role of hydrogen peroxide (H(2)O(2)) and the superoxide radical, two major ROS, in mediating those IL-1 induced responses. The results obtained show that chondrocytes produce both H(2)O(2) and superoxide radical in response to IL-1. Treatment of the chondrocyte cultures with H(2)O(2) alone did not induce NF-kappa B activation or iNOS expression. Addition of H(2)O(2) simultaneously with IL-1 did neither enhance nor inhibit NF-kappa B activation and iNOS expression, relatively to treatment with IL-1 alone. Accordingly, treatment with catalase did not inhibit those IL-1 induced responses. Treatment with superoxide dismutase, however, effectively prevented IL-1-induced I kappa B-alpha degradation and iNOS expression. Taken together, the results obtained indicate that superoxide mediates IL-1-induced I kappa B-alpha degradation and the consequent NF-kappa B activation and iNOS expression in chondrocytes, whereas H(2)O(2) does not seem to participate in those IL-1-induced responses. In conclusion, the present study identifies the superoxide radical as the ROS involved in mediating the IL-1-induced signaling pathway that leads to NF-kappa B activation and to the expression of NF-kappa B dependent genes in bovine articular chondrocytes. PMID- 12577313 TI - Vitamin D receptor is not required for the rapid actions of 1,25-dihydroxyvitamin D3 to increase intracellular calcium and activate protein kinase C in mouse osteoblasts. AB - The rapid, non-genomic actions of 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] have been well described, however, the role of the nuclear vitamin D receptor (VDR) in this pathway remains unclear. To address this question, we used VDR(+/+) and VDR(-/-) osteoblasts isolated from wild-type and VDR null mice to study the increase in intracellular calcium ([Ca(2+)](i)) and activation of protein kinase C (PKC) induced by 1,25(OH)(2)D(3). Within 1 min of 1,25(OH)(2)D(3) (100 nM) treatment, an increase of 58 and 53 nM in [Ca(2+)](i) (n = 3) was detected in VDR(+/+) and VDR(-/-) cells, respectively. By 5 min, 1,25(OH)(2)D(3) caused a 2.1 and 1.9-fold increase (n = 6) in the phosphorylation of PKC substrate peptide acetylated-MBP(4-14) in VDR(+/+) and VDR(-/-) osteoblasts. The 1,25(OH)(2)D(3) induced phosphorylation was abolished by GF109203X, a general PKC inhibitor, in both cell types, confirming that the secosteroid induced PKC activity. Moreover, 1,25(OH)(2)D(3) treatment resulted in the same degree of translocation of PKC alpha and PKC-delta, but not of PKC-zeta, from cytosol to plasma membrane in both VDR(+/+) and VDR(-/-) cells. These experiments demonstrate that the 1,25(OH)(2)D(3)-induced rapid increases in [Ca(2+)](i) and PKC activity are neither mediated by, nor dependent upon, a functional nuclear VDR in mouse osteoblasts. Thus, VDR is not essential for these rapid actions of 1,25(OH)(2)D(3) in osteoblasts. PMID- 12577314 TI - SPARC regulates cell cycle progression in mesangial cells via its inhibition of IGF-dependent signaling. AB - Glomerular mesangial cells both synthesize and respond to insulin-like growth factor-1 (IGF-1). Increased activity of the IGF signaling pathway has been implicated as a major contributor to renal enlargement and subsequent development of diabetic nephropathy. Secreted protein acidic and rich in cysteine (SPARC), a matricellular protein, has been shown to modulate the interaction of cells with growth factors and extracellular matrix. We have reported that primary glomerular mesangial cells derived from SPARC-null mice exhibit an accelerated rate of proliferation and produce substantially decreased levels of transforming growth factor beta1 (TGF-beta1) in comparison to their wild-type counterparts (Francki et al. [1999] J. Biol. Chem. 274: 32145-32152). Herein we present evidence that SPARC modulates IGF-dependent signaling in glomerular mesangial cells. SPARC-null mesangial cells produce increased amounts of IGF-1 and -2, as well as IGF-1 receptor (IGF-1R) in comparison to wild-type cells. Addition of recombinant SPARC to SPARC-null cells inhibited IGF-1-stimulated mitogen activated protein kinase (MAPK) activation and DNA synthesis. We also show that the observed accelerated rate of basal and IGF-1-stimulated proliferation in mesangial cells derived from SPARC-null animals is due, at least in part, to markedly diminished levels of cyclin D1 and the cyclin-dependent kinase (cdk) inhibitors p21 and p27. Since expression of SPARC in the glomerulus is especially prominent during renal injury, our findings substantiate previous claims that SPARC is involved in glomerular remodeling and repair, a process commonly associated with mesangioproliferative glomerulonephritis and diabetic nephropathy. PMID- 12577315 TI - Cell cycle dependent regulation of protein kinase CK2 signaling to the nuclear matrix. AB - Protein kinase CK2 is a ubiquitous protein serine/threonine kinase that is involved in cell growth and proliferation as well as suppression of apoptosis. Several studies have suggested that the kinase plays a role in cell cycle progression; however, changes in enzyme activity during phases of cell cycle have not been detected. Nuclear matrix is a key locus for CK2 signaling in the nucleus. We therefore examined CK2 signaling to the nuclear matrix in distinct phases of cell cycle by employing synchronized ALVA-41 prostate cancer cells. Removal of serum from the culture medium resulted in G0/G1 arrest, and a reduction in the nuclear matrix-associated CK2 activity which was rapidly reversed on addition of serum. Arresting the cells in G(0)/G(1) phase with hydroxyurea and subsequent release to S phase by serum gave similar results. Cells arrested in the G(2)/M phase by treatment with nocodazole demonstrated an extensive reduction in the nuclear matrix-associated CK2 which was reversed rapidly on addition of serum. Changes in the immunoreactive CK2 protein were concordant with the activity data reflecting a dynamic trafficking of the kinase in distinct phases of cell cycle. Under the same conditions, CK2 activity in total cellular lysate remained essentially unaltered. These results provide the first direct evidence of discrete modulations of CK2 in the nuclear matrix during the cell cycle progression. Inducible overexpression of CK2 in CHO cells yielded only a modest increase in CK2 activity even though a significant increase in expression was apparent at the level of CK2 alpha-specific message. Stably transfected ALVA-41 cells, however, did not show a significant change in CK2 levels despite increased expression at the message level. Not surprisingly, both types of the stably transfected cells failed to show any alteration in cell cycle progression. Distribution of the CK2 activity in the cytosolic versus nuclear matrix fractions in normal cells appears to be different from that in the cancer cells such that the ratio of nuclear matrix to cytosolic activity is much higher in the latter. Considering that nuclear matrix is central to several nuclear functions, this pattern of intracellular distribution of CK2 may have implications for its role in the oncogenic process. Published 2003 Wiley-Liss, Inc. PMID- 12577316 TI - Role of catenins in the development of gap junctions in rat cardiomyocytes. AB - Gap junctions are intercellular communicating channels responsible for the synchronized activity of cardiomyocytes. Recent studies have shown that the membrane-associated guanylate kinase protein, zonula occludens-1 (ZO-1) can bind to catenins in epithelial cells and act as an adapter for the transport of the connexin isotype, Cx43 during gap junction formation. The significance of catenins in the development of gap junctions and whether complexes between catenins and ZO-1 are formed in cardiomyocytes are not clear. In this study, immunofluorescence and confocal microscopy showed sequential redistribution of alpha-catenin, beta-catenin, ZO-1, and Cx43 to the plasma membrane when rat cardiomyocytes were cultured in low Ca(2+) (<5 microM) medium, then shifted to 1.8 mM Ca(2+) medium (Ca(2+) switch). Diffuse cytoplasmic staining of alpha catenin, beta-catenin, ZO-1, and Cx43 was seen in the cytoplasm when cardiomyocytes were cultured in low Ca(2+) medium. Staining of alpha-catenin, beta-catenin, and ZO-1 was detected at the plasma membrane of cell-cell contact sites 10 min after Ca(2+) switch, whereas Cx43 staining was first detected, colocalized with ZO-1 at the plasma membrane, 30 min after Ca(2+) switch. Distinct junctional and extensive cytoplasmic staining of alpha-catenin, beta catenin, ZO-1, and Cx43 was seen 2 h after Ca(2+) switch. Immunoprecipitation of Triton X-100 cardiomyocyte extracts using anti-beta-catenin antibodies showed that beta-catenin was associated with alpha-catenin, ZO-1, and Cx43 at 2 h after Ca(2+) switch. Intracellular application of antisera against alpha-catenin, beta catenin, or ZO-1 by electroporation of cardiomyocytes cultured in low Ca(2+) medium inhibited the redistribution of Cx43 to the plasma membrane following Ca(2+) switch. These results suggest the formation of a catenin-ZO-1-Cx43 complex in rat cardiomyocytes and that binding of catenins to ZO-1 is required for Cx43 transport to the plasma membrane during the assembly of gap junctions. PMID- 12577317 TI - Expression of interleukin-1 beta and interleukin-1 receptor antagonist in oxLDL treated human aortic smooth muscle cells and in the neointima of cholesterol-fed endothelia-denuded rabbits. AB - The migration of vascular smooth muscle cells (VSMCs) from the media to the intima and the proliferation of intimal VSMCs are key events in restenotic lesion development. These events, which are preceded and accompanied by inflammation, are modulated by the proinflammatory cytokine, interleukin-1 beta (IL-1 beta), which induces vascular smooth muscle cells to express adhesion molecules and to proliferate. IL-1 beta action is complex and regulated, in part, by its naturally occurring inhibitor, the IL-1 receptor antagonist (IL-1ra). Whether there was a temporal and spatial correlation between IL-1 beta and IL-1ra expression in, and release by, oxidized low density lipoproteins (oxLDL)-stimulated human aortic smooth muscle cells (HASMCs) was determined by using ELISA and Western blot. In addition, IL-1 beta and IL-1ra expression was detected in the neointima of endothelia-denuded cholesterol-fed New Zealand white rabbits by immunohistochemistry and Western blot. In HASMCs, oxLDL induced IL-beta and IL 1ra expression and release in a dose- and time-dependent manner. Treatment with 20 microg/ml oxLDL resulted in increased IL-1 beta release after 6 h, which peaked at 24 h, and in increased IL-1ra release, first seen after 12 h, but continuing to increase for at least 48 h. In the cells, IL-beta expression showed a similar pattern to release, whereas IL-1ra expression was seen in unstimulated cells and was not increased by oxLDL treatment. Confocal microscopy showed colocalization of IL-beta and IL-1ra expression in oxLDL-stimulated HASMCs. oxLDL caused significant induction of nuclear factor kappa B and activator protein-1 DNA binding activity in HASMCs (6.6- and 3.3-fold, respectively). In cholesterol fed endothelia-denuded rabbits, the notably thickened intima showed significant IL-1 beta and IL-1ra expression. These results provide further support for the role of IL-1 system in the pathogenesis of restenosis. This is the first demonstration of IL-1 beta and IL-1ra expression and secretion of oxLDL-treated HASMCs and their expression in the rabbit neointima, suggesting that the smooth muscle cells of the intima are an important source of these factors. PMID- 12577318 TI - Calcyclin (S100A6) regulates pulmonary fibroblast proliferation, morphology, and cytoskeletal organization in vitro. AB - Calcyclin (S100A6) is a member of the S100A family of calcium binding proteins. While the precise function of calcyclin is unknown, calcyclin expression is associated with cell proliferation and calcyclin is expressed in several types of cancer phenotypes. In the present study, the functional role of calcyclin was further elucidated in pulmonary fibroblasts. Antisense S100A6 RNA expression inhibited serum and mechanical strain-induced fibroblast proliferation. This attenuated proliferative response was accompanied by a flattened, spread cell morphology, and disruption of tropomyosin labeled microfilaments. Changes in cytoskeletal organization did not correspond with a decrease in tropomyosin levels. These observations suggest a role for calcyclin in modulating calcium dependent signaling events that regulate progression through the cell cycle. J. Cell. Biochem. 88: 848-854, 2003. PMID- 12577319 TI - Intraneuronal localization of Nogo-A in the rat. AB - Nogo-A is known to be a myelin-associated protein with strong inhibitory effect on neurite outgrowth and has been considered one of the major factors that hinder fiber regeneration in the central nervous system. Recent studies have demonstrated widespread occurrence of nogo-A mRNA and Nogo-A protein in neurons. Our concurrent immunohistochemical study substantiated the widespread distribution of neuronal Nogo-A. The present study was thus focused on its intraneuronal distribution in the central nervous system, using Western blotting, immunohistochemical, and immunogold electron microscopic techniques. Western blotting of the nucleus, cytoplasm, and membrane subcellular fractions of the cerebellum and spinal cord tissues demonstrated that all three fractions contained Nogo-A. Nogo-A immunoreactivity could be identified under confocal microscope in the nucleus, perikayon, and proximal dendrite and along the cell membrane. Under the electron microscope, the perikaryonal Nogo-A immunogold particles were mainly distributed at polyribosomes and rough endoplasmic reticulum, suggesting its relationship with translation process. The immunogold particles could also be found beneath or on the plasma membrane. In the nucleus, the Nogo-A immunogold particles were found to be localized at the chromatins of the nucleus, indicating its possible involvement in gene transcription. The presence of Nogo-A in the nucleus was further supported by transfection of COS-7L cells with nogo-A. This study provides the first immunocytochemical evidence for intraneuronal distribution of Nogo-A. Apparently, the significance of Nogo-A in the central nervous system is far more complex than what has been envisioned. PMID- 12577321 TI - Distribution of dopamine D2 receptor mRNAs in the brain and the pituitary of female rainbow trout: an in situ hybridization study. AB - The distribution of D(2)R (dopamine D(2) receptor) mRNAs was studied in the forebrain of maturing female rainbow trout by means of in situ hybridization using a (35)S-labeled riboprobe (810 bp) spanning the third intracytoplasmic loop. A hybridization signal was consistently obtained in the olfactory epithelium, the internal cell layer of the olfactory bulbs, the ventral and dorsal subdivisions of the ventral telencephalon, and most preoptic subdivisions, with the notable exception of the magnocellular preoptic nucleus, and the periventricular regions of the mediobasal hypothalamus, including the posterior tuberculum. In the pituitary, the signal was higher in the pars intermedia than in the proximal and the rostral pars distalis, but no obvious correspondence with a given cell type could be assigned. Labeled cells were also located in the thalamic region, some pretectal nuclei, the optic tectum, and the torus semicircularis. These results provide a morphologic basis for a better understanding on the functions and evolution of the dopaminergic systems in lower vertebrates. PMID- 12577320 TI - Parvalbumin, calbindin, or calretinin in cortically projecting and GABAergic, cholinergic, or glutamatergic basal forebrain neurons of the rat. AB - The basal forebrain (BF) plays an important role in modulating cortical activity and facilitating processes of attention, learning, and memory. This role is subserved by cholinergic neurons but also requires the participation of other noncholinergic neurons. Noncholinergic neurons include gamma-amino butyric acidergic (GABAergic) neurons, some of which project in parallel with the cholinergic cells to the cerebral cortex, others of which project caudally or locally. With the original aim of distinguishing different subgroups of GABAergic neurons, we examined immunostaining for the calcium binding proteins (CBPs) parvalbumin (Parv), calbindin (Calb), and calretinin (Calret) in the rat. Although the CBP(+) cell groups were distributed in a coextensive manner with the GABAergic cells, they were collectively more numerous. Of cells retrogradely labeled with cholera toxin (CT) from the prefrontal or parietal cortex, Parv(+) and Calb(+) cells, but not Calret(+) cells, represented substantial proportions ( approximately 35-45% each) that collectively were greater than that of GABAergic projection neurons. From dual immunostaining for the CBPs and glutamic acid decarboxylase (GAD), it appeared that the vast majority (>90%) of the Parv(+) group was GAD(+), whereas only a small minority (<10%) of the Calb(+) or Calret(+) group was GAD(+). Significant proportions of Calb(+) (>40%) and Calret(+) (>80%) neurons were immunopositive for phosphate-activated glutaminase, the synthetic enzyme for transmitter glutamate. The results suggested that, whereas Calret(+) cells predominantly comprise caudally or locally projecting, possibly glutamatergic BF neurons, Parv(+) cells likely comprise the cortically projecting GABAergic BF neurons and Calb(+) cells the cortically projecting, possibly glutamatergic BF neurons that would collectively participate with the cholinergic cells in the modulation of cortical activity. PMID- 12577322 TI - Penile neuronal nitric oxide synthase and its regulatory proteins are present in hypothalamic and spinal cord regions involved in the control of penile erection. AB - Control of penile erection requires the coordination of the hypothalamus and the L6-S1 region of the spinal cord. Erection requires the activation of neuronal nitric oxide synthase (nNOS), which is tightly regulated. Because variants of nNOS (penile nNOS: PnNOS) and the N-methyl-D-aspartate receptor (truncated NMDAR subunit 1: NMDAR1-T) as well as protein inhibitor of NOS (PIN) have all been located in the pelvic ganglia and penile nerves, this work aims to determine whether these proteins are also present in the hypothalamus. It was found that PnNOS, the brain-type nNOS, and PIN, were expressed in the hypothalamus. In contrast, NMDAR1-T was expressed only in the penis, whereas the brain-type NMDAR1 was present in the brain and sacral spinal cord and not in the penis. PnNOS was found in the media preoptic area, posterior magnocellular, and the parvocellular regions of the paraventricular nucleus, supraoptic nucleus, septohypothalamic nucleus, medial septum, cortex, and in some of the nNOS staining neurons throughout the brain. It was absent in the organum vasculosum of the lamina terminalis. PIN staining was present in neurons of the medial preoptic area, paraventricular nucleus, medial septum, and cortex, but not in the supraoptic nucleus, septohypothalamic nucleus, or organum vasculosum of the lamina terminalis. Colocalization between PnNOS and PIN was found in the medial preoptic area, medial septum, and cortex, and less in the paraventricular nucleus. PnNOS and oxytocin were colocalized in the paraventricular nucleus and supraoptic nucleus. In hypothalamic extracts, recombinant PIN-GST protein bound to PnNOS in the extracts and partially inhibited NOS activity. These results indicate that both nNOS variants, and their respective regulatory proteins are present and colocalize in the hypothalamic and spinal cord regions involved in penile erection. PMID- 12577323 TI - Presynaptic neurotrophin-3 increases the number of tectal synapses, vesicle density, and number of docked vesicles in chick embryos. AB - To determine whether presynaptically derived neurotrophins may contribute to synaptic plasticity, we examined whether neurotrophin-3 (NT-3) changed the number, size, vesicle content, or vesicle distribution of synapses within the retinorecipient layers of the chick optic tectum. In this system, endogenous NT-3 derives presynaptically from retinal ganglion cell axons. Retinotectal synapses comprise the majority of synapses in superficial tectal layers, as demonstrated by destruction of retinotectal input by intraocular application of the drug monensin. To examine the effect of increased or decreased levels of NT-3, either exogenous NT-3 or monoclonal NT-3 blocking antibodies were injected into the optic tectum of 19-day-old chick embryos, spiked with radiolabeled protein to verify the success of injections and estimate effective concentrations. After 48 hours, the ultrastructure of superficial tectal layers was analyzed and compared with samples from control tecta injected with cytochrome C. NT-3 increased the number of synapses, synaptic vesicles/profile, synaptic vesicle densities, the number of docked vesicles, and the length of the synaptic profile. Deprivation of anterogradely transported endogenous NT-3 with NT-3 antibodies resulted in the opposite effect: decreased numbers of synapses, decreased vesicle densities, and decreased numbers of docked vesicles. Brain-derived neurotrophic factor (BDNF) had a largely different effect than NT-3. BDNF increased the density of vesicles and deprivation of endogenous TrkB ligands with TrkB fusion protein reduced the density of vesicles in the synapses, without effects on synapse number or docked vesicles. We conclude that anterogradely transported NT-3 affects synapse strength in a way that differs from that of presumably postsynaptic-derived BDNF. PMID- 12577324 TI - Morphologic and cytochemical criteria for the identification and delineation of individual subnuclei within the lateral habenular complex of the rat. AB - The lateral habenular complex is part of the habenular nuclei, a distinct structure in the dorsal diencephalon of all vertebrates. In contrast to the bewildering diversity of behaviors, in which the lateral habenular complex is thought to be involved, there is an astonishing lack of information concerning its cellular organization, its neuronal circuits, and the neurophysiological mechanisms, which may provide the physiological and molecular basis for its diverse biological functions. This problem may be due to an unexpected heterogeneity of the lateral habenular complex. Recently, a detailed subnuclear organization has been described (Andres et al. [1999] J Comp Neurol 407:130-150), which provides the base for a subsequent physiological and behavioral analysis of this area. Available criteria, however, can be applied to semithin sections only. To facilitate further investigations, the present work aimed to elaborate novel morphologic and immunocytochemical criteria that can be applied to conventional cryostat or Vibratome sections to allow identification and delineation of subnuclei of the lateral habenular complex. Consequently, the regional, cellular, and subcellular localization of approximately 30 different neuroactive molecules was investigated. Of these candidate molecules, gamma-aminobutyric acid-B receptor protein, Kir3.2 potassium channel protein, tyrosine hydroxylase, and neurofilament heavy chain proved to be suitable markers. Our observation suggests that the habenular subnuclei express distinct immunocytochemical characteristics. These features may be used to identify and delineate the subnuclei on conventional cryostat or Vibratome sections. From our results, it is expected that the further functional analysis of the lateral habenular complex will be facilitated considerably. PMID- 12577325 TI - Role of EphA4 in defining the position of a motoneuron pool within the spinal cord. AB - The correct assembly of the neural circuits that control movement requires the development of topographically organized pools of motoneurons within the spinal cord. The generation of a diverse array of motoneuron subtypes, which express differing transcription factors and cell-surface receptors, allows different motoneuron pools to be segregated to specific positions during development. In this investigation, we show that the Eph receptor tyrosine kinase, EphA4, appears to be important for the correct localization of a motoneuron pool to a specific position in the spinal cord. In the spinal cord of mice deficient in EphA4, the motoneuron pool that innervates the tibialis anterior muscle of the hindlimb is caudally displaced by approximately one vertebral segment. However, despite the abnormal position of the tibialis anterior motoneuron pool in the spinal cord of EphA4-deficient animals, the motoneurons of this pool still project to the tibialis anterior muscle of the hindlimb correctly. Additional analyses of other limb innervating motoneuron pools in the cervical and lumbar enlargements of the spinal cord of EphA4-deficient animals revealed them to be located in the appropriate segmental positions. Furthermore, we show that EphA4 does not appear to be important for spinal motoneuron survival as stereological quantification of the number of motoneurons present in the sciatic motoneuron pool of EphA4 deficient animals demonstrated these motoneurons to be present in the correct numbers. These observations suggest an important role for EphA4 in regulating the position of a specific motoneuron pool within the spinal cord. PMID- 12577326 TI - [Revisiting experiences of integrated traditional Chinese and Western medicine]. PMID- 12577327 TI - [A controlled clinical study between hepatic arterial infusion with embolized curcuma aromatic oil and chemical drugs in treating primary liver cancer]. AB - OBJECTIVE: To evaluate the effectiveness, toxicity and prospective application of hepatic arterial infusion (HAI) with Embolized Curcuma Aromatic oil (CAO) in treating primary liver cancer (PLC). METHODS: In the treated group, 32 patients with PLC were treated by HAI with 1-3 ml of embolized CAO and oral administration of Chinese herbal medicine. In the control group, 32 patients with PLC were treated with transcatheter artery chemoembolization (TACE). RESULTS: In the treated group, one patient attained complete remission (CR) and 13 partial remission (PR), the total effective rate being 43.75%. The level of alpha fetal protein (AFP) turned to normal range in 7 cases and decreased in other 7. In the control group, 10 obtained PR and the total effective rate being 31.25%, AFP level turned to normal in 5 and decreased in 2. There was no statistical significance between the two groups. The incidences of post-embolism syndrome, such as fever, abdominal pain and vomiting were similar between the two groups but no myelosuppression occurred in the treated group with significant difference (P < 0.01) as comparing with that in the control group. The mean survival time, median survival time, 1-, 2- and 3-year survival rate in the treated group was 11.5 months, 10 months, 37.5%, 13.3% and 6.9% respectively, while in the control group was 7.25 months, 6 months, 15.6%, 3.2% and 0 respectively. The treated group was better in mean survival time, median survival time and 1-year survival rate than that of the control group (P < 0.05). CONCLUSION: HAI with embolized CAO showed a similar favorite effect in treating PLC as that of TACE, but superior than TACE with longer survival time and milder myelosuppression. PMID- 12577328 TI - [Clinical study on treatment of middle-advanced stage liver cancer by combined treatment of hepatic artery chemoembolization with gan'ai no. I and no. II]. AB - OBJECTIVE: To observe the clinical effect of combined treatment of hepatic artery chemoembolization (HACE) and Chinese herbal medicine (CHM) in treating middle advanced stage liver cancer. METHODS: Sixty patients with middle-advanced stage liver cancer were randomly divided into two groups. The 30 patients in Group A were treated with combined HACE and Chinese herbal medicine (Gan'ai No. I and No. II) and the other 30 in Group B were treated with HACE alone. All patients were followed up for over 3 years. RESULTS: The 0.5-, 1- and 2-year survival rate in Group A was 76.7%, 56.7% and 30.0% respectively, and those in Group B was 50.0%, 33.3% and 16.7% respectively. The 1- and 2-year recurrence rate in Group A was 43.3%, 66.7% and that in Group B was 66.7%, 90.0% respectively. Moreover, Group A was significantly superior to Group B in tumor shrinking, AFP decreasing and blood leucocyte reducing (P < 0.01), as well as in improving clinical symptoms. CONCLUSION: The combined treatment has obvious effect in treating middle-advanced stage liver cancer. PMID- 12577329 TI - [Effect of huangqi zengmian powder on interstitial response in patients with esophageal cancer at peri-operational period]. AB - OBJECTIVE: To observe the effect of Huangqi Zengmian Powder (HQZMP) on interstitial response of esophageal cancer at peri-operational period. METHODS: Thirty-seven patients of esophageal cancer were treated with HQZMP for 4 weeks peri-operationally. The pathological interstitial changes (including mastocyte, microvessels and lymphocytic response) in resected esophageal cancer sample were observed using toluidine blue stain. RESULTS: As compared with the non-HQZMP treated control group, quantity of interstitial mastocyte, focal lymphocytic aggregation and interstitial microvessel damage in the HQZMP treated group were more obvious (P < 0.01). CONCLUSION: HQZMP could improve the immune function of esophageal patients in peri-operational period. PMID- 12577330 TI - [Clinical observation on effect of electroacupuncture therapy in treating superficial tumor]. AB - OBJECTIVE: To investigate the clinical effect of electroacupuncture therapy (EAT) in treating superficial tumors. METHODS: The healthy tissue was protected by insulation sleeve, and the platinum electrodes served as needles was inserted into the tumor and connected to an EAT instrument using galvanic current. The electric voltage applied was 6-8 V, the electric current was in a range of 40-80 mA, and 80-100 coulomb electricity for 1 cm diameter of tumor mass was administered. RESULTS: In the 320 cases, 123 were complete remission (CR), 129 partial remission (PR), 36 with their tumor shrinked by 1/4 and 32 with size of tumor unchanged. The total effective rate (CR + PR) was 78.7%. CONCLUSION: EAT shows good effect in treating superficial tumor and provides a new therapeutic means for the patients with tumor of unresectable or relapsed. It is a simple, convenient, safe and effective method with less injury and quick recovery. PMID- 12577331 TI - [Clinical observation on gastrointestinal protective effect of Composite Salviae injection in patients undergoing cardio-pulmonary bypass heart surgery]. AB - OBJECTIVE: To observe the gastrointestinal protective effect of Composite Salviae Injection (CSI) in patients undergoing cardio-pulmonary bypass (CPB) heart surgery. METHODS: Eighteen patients, who were scheduled to cardiac surgery (either atrial or ventrical septal repairing) undergoing CPB were randomized equally into two groups. Before CPB, the CSI group was treated with CSI 0.5 ml/kg by intravenous dripping and the control group was treated with normal saline in equal volume. The intragastric mucosa pH value (pHi) of patients was monitored by tensiometer. RESULTS: As compared with the pre-CPB value, pHi lowered significantly during, 1 h and 2 hrs after CPB in the control group (P < 0.05 or P < 0.01), while in the CSI group, pHi lowered significantly only during CPB (P < 0.05), but with insignificant change at 1 h and 2 hrs after CPB. Comparison between the two groups showed that pHi value in the CSI group was higher significantly than that in the control group at all respective monitoring period (P < 0.05). CONCLUSION: CSI has gastrointestinal protective effect in patients undergoing CPB cardiosurgery to some extent. PMID- 12577332 TI - [Effects of combined transdermal acupoint electric stimulation and isoflurane anesthesia on isoflurane-induced hypotension and tissue oxygen metabolism in patients undergoing craniotomy]. AB - OBJECTIVE: To observe the effects of combined transdermal acupoint electric stimulation and isoflurane anesthesia on isoflurane-induced hypotension and tissue oxygen metabolism in patients undergoing craniotomy. METHODS: Forty-two patients with brain tumor of ASA physical status I or II and scheduled for elective surgery were randomly divided into two groups. The craniotomy was undergoing in Group A with isoflurane anesthesia, while in Group B, with combined transdermal acupoint electric stimulation (TAES) and isoflurane anesthesia. Isoflurane induced hypotension was performed in both groups by augmenting the concentration of isoflurane to lower the mean arterial pressure than before anesthesia for 30%-40% and maintain for 30-45 mins. The tissue oxygen metabolism, blood gas and arterial lactic acid level before, during and after hypotension were monitored by Swan-Ganz floating catheterization. RESULTS: As compared with before hypotension, the pulmonary arterial and mixed with venous blood oxygen saturation lowered and oxygen supply lowered, and oxygen uptake increased significantly in both groups (P < 0.05), but the extent of changes in oxygen metabolic criteria in Group B were all lesser than those in Group A (P < 0.05). CONCLUSION: Combined acupoint electric stimulation and general anesthesia could maintain the tissue oxygen supply and demand balance better in the isofluraneinduced hypotension process. PMID- 12577333 TI - [Effect of combination of ligustrazine and nitrendipine on hemodynamics in patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To find the drugs for lowering the pulmonary vascular resistance in patients of chronic obstructive pulmonary disease (COPD) but would not increase its side-effect. METHODS: Changes of some hemodynamic parameters in 15 patients of COPD in stable stage treated by nitrendipine and ligustrazine were observed by right heart floating catheterization. RESULTS: The mean pulmonary arterial pressure reduced by 1.4% immediately after the combined treatment, and reduced by 17.0%, 20.0% and 18.0% in 15 mins, 30 mins and 60 mins after treatment. In the responding time, the pulmonary vascular resistance reduced by 15.2%, 36.2%, 43.0% and 34.6%, the systemic vascular resistance reduced by 7.9%, 19.2%, 17.9% and 20.8%, and the cardiac output increased by 15.8%, 22.6%, 22.2%, and 33.8% respectively. The changes of systemic arterial pressure and heart rate were of no statistical significance. CONCLUSION: Combination therapy of nitrendipine and ligustrazine could lower the pulmonary arterial pressure and pulmonary vascular resistance effectively and has a selective effect on pulmonary circulation in patients with COPD. PMID- 12577334 TI - [Changes of segmental motion after spinal manipulation in patients with lumbar intervertebral disc protrusion]. AB - OBJECTIVE: To explore the effect of manipulation on segmental motion of lumbar vertebra and the clinical significance before and after treatment, and evaluation with charge couple device (CCD) measurement system. METHODS: CCD and dynamic radiography were conducted in 30 patients before and after treated by manipulation or other conservative therapy, analysis and comparisons in segmental motion ranges of flexion-extension and lateral bending, correlation between data of CCD system and radiography as well as characteristics of lumbar vertebral function recovery after manipulation were performed. RESULTS: Certain correlation between data of CCD and radiography was shown by linear correlation test (r = 0.42-0.72, P < 0.05-0.01). Paired t test showed that the lower lumbar segmental flexion-extension function was significantly recovered (t = 2.26-2.61, P < 0.05 0.01) after manipulation, but the recovery on other segments was insignificant. CONCLUSION: The special designed CCD system is of important clinical significance in assessing segmental lumbar vertebral function. Lower lumbar vertebral flexion extention functional state is a crucial criterium for evaluation of manipulative therapy in lumbar intervertebral disc protrusion. PMID- 12577335 TI - [Study on reference laboratory diagnostic index of liver-fire ascending syndrome]. AB - OBJECTIVE: To explore the binding point of Syndrome Differentiation of TCM and disease differentiation by western medicine. METHODS: Selected laboratory diagnostic indexes were tested in 59 cases of Liver-Fire Ascending Syndrome (LFAS). Referring to clinical typing and staging, the data obtained were analysed by same syndrome with different diseases and same disease with different syndrome as well as the values determined in related syndrome-types were compared. RESULTS: The levels of prostaglandin F2 alpha (PGF2 alpha) and arginine vasopressin (AVP) in patients of LFAS were higher than those in patients with Liver-Kidney Yin-Deficiency Syndrome and Liver-Yang Hyperactivity Syndrome. As for linking between Syndrome-type and stage of disease, most cases (70%) of herpes simplex viral keratitis of LFAS were deep matrix layer type; and 100% hypertension patients with LFAS belonged to stage II. CONCLUSION: PGF2 alpha and AVP could be used as the reference laboratory diagnostic indexes of LFAS. PMID- 12577336 TI - [Preliminary experimental study on therapeutic mechanism of kangzhen zhijing capsule in treating Parkinson's disease]. AB - OBJECTIVE: To explore the therapeutic mechanism of Kangzhen Zhijing Capsule (KZZJC) on rats with Parkinson's disease (PD). METHODS: PD rat model was established by microinjection of 6-hydroxydopamine solution into left substantia nigra zona compacta (SNZC). Model rats were randomly divided into 3 groups and treated with KZZJC, levodopa and normal saline respectively for 30 days. The rotational behavior of rats before and after treatment was observed. Then rats were sacrificed and relevant indexes of left SNZC and striatum were examined. RESULTS: In rats treated with KZZJC, the rotational behavior was improved (P < 0.05), the levels of dopamine (DA) content and DA/homovanillic acid (HVA) ratio in striatum, and glutathione peroxidase, superoxide dismutase and glutathione in SNZC increased, but malondialdehyde in SNZC decreased, as compared with those in rats treated with normal saline, the difference was significant (P < 0.01 or P < 0.05). Rotational behavior in rats treated by levodopa was improved significantly (P < 0.01). CONCLUSION: KZZJC could partially improve the rotational behavior in PD model rats, increase DA content and DA/HVA ratio in striatum. The mechanism might be the reducing of free radical damage and DA catabolism. PMID- 12577337 TI - [Study on effect of Puerarin on nitric oxide system in rats' tissue and its mechanism]. AB - OBJECTIVE: To evaluate the effect of Puerarin on nitric oxide (NO) content and nitric oxide synthase (NOS) activity in rats' tissue. METHODS: Sixteen adult Wistar rats were randomly divided into two groups, 8 in each group. The Purerain group was intraperitoneal administered Puerarin 80 mg/kg.d and the control group was given propylene glycol by intraperitoneal injection for 30 days. The NO content in rats' heart, brain, liver and kidney was determined by nitratase method and NOS was detected by oxyhemoglobin oxidation method. Besides, the electromicroscopic examination on rats' liver and brain was also conducted. RESULTS: NO content in heart, brain, liver and kidney in the Puerarin group was significantly higher than that in the control group respectively (P < 0.05 or P < 0.01). NOS was detected only in heart and brain in both groups and it was also higher in the Puerarin group than that in the control group (P < 0.05). No abnormal ultrastructure was detected by electromicroscope in liver and brain of both groups. CONCLUSION: Increasing NO content and NOS activity in tissues may be one of the mechanism for pharmaceutical action of Puerarin. PMID- 12577338 TI - [Dynamic observation on imbalance of dynamic and static forces and degenerated cervical intervertebral disc in rats]. AB - OBJECTIVE: To study the relation between biomechanical imbalance and the degree and course of degeneration of cervical intervertebral disc in rats. METHODS: Sixty SD rats, 8 months old, were randomly divided into 6 groups, the control and model groups of 3, 5, 7 months, 10 in each group. The cervical disc dynamic and static forces imbalance of degeneration model was established to assess the degree of degeneration as well as the content some inflammatory mediators (prostaglandin E2, 6-keto-prostaglandin F1 alpha) and collagenase (MMP-1) activity. RESULTS: (1) In 3 months model group and 5 months control group, fibrous ring of intervertebral disc showed some fissure and slight irregular arrangement, nucleus pulposus shrunken or became smaller, mild herniation of nucleus pulposus was seen in some disc. The nucleus pulposus of intervertebral disc in 5 months model group was fibrosed completely and the disc in 7 months model group herniated or became osteophytosis. (2) Compared with the control group of same time period, MMP-1 was increased significantly in the 5 months and 7 months model groups (P < 0.05), and prostaglandin E2 and 6-keto-prostaglandin F1 alpha elevated in the model groups (P < 0.05, P < 0.01). (3) Comparison between model groups, MMP-1 activity in 5 months and 7 months groups was higher than that in the 3 months group (P < 0.05), and 6-keto-prostaglandin F1 alpha was higher in 7 months group than that in 3 months group (P < 0.05). CONCLUSION: Cervical intervertebral disc undergoes a progressive degenerating process. By breaking the dynamic-static balance of neck in rats could accelerate the progression of degeneration. The fact could be used to elucidate the theory of pathogenesis of cervical spondylopathy, dynamic force imbalance in priority and static force imbalance in predominance. PMID- 12577339 TI - [Effect of guizhi decoction on adenyl cyclase and phosphodiesterase in hypothalamus of rats models of fever and hypothermia]. AB - OBJECTIVE: To observe the effect of Guizhi Decoction (GZD) on adenyl cyclase (AC) and phosphodiesterase (PDE) activities in hypothalamus of rat models of fever and hypothermia. METHODS: The AC and PDE activities in hypothalamus were determined using radio-isotope method. RESULTS: GZD could lower the AC activity in yeast induced fever rat model (P < 0.05), but cause rise of AC activity in aminopyrine induced hypothermia model (P < 0.05). No significant influence of GZD on PDE activity was shown in both fever or hypothermia models. CONCLUSION: The bi directional thermo-regulation effect of GZD might be partially due to influence on AC activity. PMID- 12577340 TI - [Mechanism of sini decoction in regulating beta-adrenergic receptor signal transduction in rats with myocardial ischemia]. AB - OBJECTIVE: To investigate the effect of Sini Decoction (SND) on beta-adrenergic receptor (beta-AR) signal transduction in rats with myocardial ischemia. METHODS: After beta-AR had been blocked by propranolol, the myocardial ischemia rats were established by pituitrin injection. The density of beta-AR in myocardial cell membrane was determined by radioligand binding assay, cAMP level in plasma and myocardial tissue was detected by radioimmunoassay (RIA), and the mRNA expression of beta 1-AR and beta 1-AR kinase (beta ARK-1) was measured by quantitative reverse transcriptase polymerase chain reaction (RT-PCR) amplifying assay. RESULTS: In myocardial ischemia group, the density of beta 1-AR was up-regulated and the mRNA expression of beta ARK-1 increased (P < 0.01), but the cAMP level in plasma and myocardium was lower than that in the normal group (P < 0.01 and P < 0.05). SND could promote beta 1-AR mRNA expression and inhibit beta ARK-1 mRNA expression so as to increase the density of beta-AR and cause significant raising of cAMP level in plasma and myocardium. CONCLUSION: SND could reduce the desensitization of beta 1-AR during myocardial ischemia and improve signal transduction of beta-AR. PMID- 12577341 TI - [Functional and morphological observation on cells in allergic rhinitis guinea pigs with cold or heat syndrome]. AB - OBJECTIVE: To investigate the functional and morphological characteristics of cells in allergic rhinitis (AR) guinea-pigs with Cold or Heat Syndrome. METHODS: The Cold Syndrome or Heat Syndrome guinea-pig model was made, and then reformed into AR model or rhinitis medicamentosa (RM) model by using nose drops of 10% toluene di-isocyanate (TDI) or 0.1% naphazoline hydrochloride. The cell function, count and ultramicroscopic morphology characteristics in the model animals were observed. RESULTS: The degranulation index (DI), of blood basophils and the count of mast cells (MC) in nasal mucosa were higher in AR guinea-pigs with Cold Syndrome than that in Heat Syndrome guinea-pigs, but the two parameters in AR with both Cold and Heat Syndrome were higher than those in the RM model. Mast cell and eosinophilic granulocyte in AR guinea-pigs with Cold Syndrome were high in count, widely infiltrated, indiscernible cellular capsule, heterogeneous in density, and vacuoles, fusions and tubules in granules while in those with Heat Syndrome, mast cells and eosinophils were scarcely found, without infiltration, granules with discernible membrane and homogeneous in density, no vacuoles, fusions and tubules in granules. CONCLUSION: The basophilic cells, mast cells and eosinophilic granulocytes in AR guinea-pigs with Cold Syndrome are in activated condition. PMID- 12577342 TI - [Clinical significance of study on leucocyte rheology]. PMID- 12577343 TI - [Progress in research on anti-relapsing mechanism of Chinese herbal medicine in the treatment of peptic ulcer]. PMID- 12577344 TI - [Studies on male reproductive regulation by Chinese herbal medicine]. PMID- 12577345 TI - [Summary of National Academic Conference on emmeniopathy and infertility of integrated traditional Chinese and Western medicine]. PMID- 12577346 TI - [Survey of the treatment of psoriasis by integrated traditional Chinese and Western medicine]. PMID- 12577347 TI - [Study on microcirculation of blood stasis syndrome]. PMID- 12577348 TI - [Integrated traditional Chinese and Western medicine]. PMID- 12577349 TI - [Observation on collaborative treatment of dilated cardiomyopathy]. AB - OBJECTIVE: To observe the therapeutic effect of integrated traditional Chinese and western medicine (TCM-WM) in treating dilated cardiomyopathy (DCM). METHODS: Patients of DCM were randomly divided into two groups, the 164 patients in the TCM-WM group were treated with combination therapy of traditional Chinese and western medicine, consisting of conventional western medicine, such as cardiac diuretic, vasodilative agents, taurine, coenzyme Q10, antiarrhythmics, beta blockers and Chinese herbal preparations such as Astragalus membranaceus and Shengmai injection; while the 156 cases in the control group were treated with conventional western medicine alone, including polarized liquid therapy, etc. RESULTS: The improvement of clinical symptoms and heart function in the TCM-WM group was significantly better than that in the control group. Although the total number of deceased cases in the two groups were similar, the dead number in 3-6 months in the TCM-WM group was less than that in the control group. Moreover, 1 year later, the deceased number of patients insisted with TCM-WM treatment for over 1 year was significantly less than in those treated for only 3-6 months (1 case vs 11 cases). CONCLUSION: Under the condition that there is no specific effective drugs, TCM-WM therapy can yet be regarded as an acceptable therapy for treatment of dilated cardiomyopathy. PMID- 12577351 TI - [Clinical study on effect of ginsenoside in inducing rectal cancer cell apoptosis]. AB - OBJECTIVE: To explore the effect of ginsenoside in inducing rectal cancer cell apoptosis. METHODS: Fifty patients of rectal cancer diagnosed by pathological examination were randomly divided into two groups, 35 in the treated group and 15 in the control group. The treated group received retention enema with 84.5% ginsenoside for 4-6 hrs every day, 6-8 days consecutively before surgical operation. The control group also received retention enema in the same way but with normal saline instead of ginsenoside. Fresh sample was taken during the operation and examined by electron microscopy. RESULTS: Symptoms, such as frequent defecation, hematochezia and tenesmus, were palliated in most patients (25/35) and abdominal pain relieved in all the 7 cases of incomplete intestinal obstruction in the treated group. Electron microscopic examination showed that cell apoptosis was observed in cancer sample of 23 cases among the 35 cases of the treated group, amounting to 65.7%. While in the control group, above mentioned changes were not observed at all. CONCLUSION: Ginsenoside has the effect of inducing apoptosis in rectal cancer patients. PMID- 12577350 TI - [Clinical observation on effect of intervention therapy with anticoagulant in treating patients of hypertension complicated with acute ischemic cerebrovascular disease]. AB - OBJECTIVE: To study the changes of blood coagulation-fibrinolysis-C protein system in patients of hypertension with acute ischemic cerebrovascular disease (AICVD) after anticoagulation therapy. METHODS: Fifty-seven cases of AICVD were grouped and treated with heparin and Xueshuantong (XST) respectively, and the levels of prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), tissue plasminogen activator (t-PA), plasminogen activator inhibitor (PAI) and activated protein C (APC) were determined before and after treatment. RESULTS: The clinical markedly effective rate in the heparin group and the XST group was 69.2% and 67.7% respectively with no significant difference (P > 0.05). In the heparin group, level of PT and APTT prolonged, FIB decreased, t PA activity elevated and PAI activity lowered, and APC unchanged. In the XST group, same changes in FIB, t-PA and PAI were shown but with APC increased, and PT and APTT unchanged. CONCLUSION: Both heparin and XST have good anticoagulatory function and show good clinical effect in treating patients with hypertension complicated with acute ischemic cerebrovascular diseases. PMID- 12577352 TI - [Study on effect of moxibustion and guben yiliu III combined with chemotherapy in treating middle-late stage malignant tumor]. AB - OBJECTIVE: To observe the complementary function of moxibustion and Guben Yiliu III (GBYL), a Chinese herbal composite preparation, in combination with chemotherapy. METHODS: Eighty-one patients of middle-late stage malignant tumor were randomly divided into three groups, 16 in the control group treated with chemotherapy alone, 35 in the TCM group treated with chemotherapy combined GBYL and 30 in the TCM combined moxibustion group. The therapeutic effect of treatment was evaluated according to the immediate effect, living quality scoring, etc. RESULTS: The comprehensive living quality score and Karnofsky score dropped significantly in the control group after treatment with significant increase of pain score and decrease of tongue figure score (P < 0.05 or P < 0.01). In the TCM group, the former two criteria lowered less than those in the control group, but no significant difference in the latter two criteria was shown. For those in the TCM combined moxibustion group, all the four criteria were improved better than those in the control group (P < 0.01 or P < 0.05). CONCLUSION: GBYL combined or not combined with moxibustion could improve the living quality of patients received chemotherapy. PMID- 12577354 TI - [Clinical study on effect of zhuhuang granule no. 2 in treating psoriasis with liver-qi stagnancy]. AB - OBJECTIVE: To observe the efficacy of Zhuhuang Granule No. 2 (ZHG2) in treating psoriasis with Liver-Qi stagnancy. METHODS: Sixty-seven patients were randomly divided into two groups, the 33 patients in the control group were treated with composite indigo capsule and the 34 in the treated group treated with ZHG2, and the clinical effect, changes of main symptoms and local skin lesion after treatment were evaluated. Meanwhile, the plasma levels of substance P (SP) and vasoactive intestinal peptide (VIP) in 15 patients and 13 healthy subjects were measured using radioimmunoassay (RIA). RESULTS: The total effective rate in the treated group and the control group was 97.1% and 78.8% respectively with significant difference, P < 0.05. ZHG2 showed a significant effect in reducing plasma level of SP and VIP in patients of psoriasis with Liver-Qi stagnancy. CONCLUSION: The effect of ZHG2 in treating psoriasis patients with Liver-Qi stagnancy is satisfactory, and worth further studying. PMID- 12577353 TI - [Clinical study on effect of gushukang granule in preventing and treating primary osteoporosis]. AB - OBJECTIVE: To investigate the effect and mechanism of Gushukang granule (GSKG), a Chinese herbal preparation, in preventing and treating primary osteoporosis. METHODS: The total of 197 patients of osteoporosis, whose bone mineral density (BMD) was below 1 standard deviation of the peak bone mass of same sex, with no complications and had not been treated were selected. Among them, 176 were menopausal females for over 1 year and 21 males aged over 70. The females were voluntarily divided into three groups, the 101 cases in the observed group were treated with GSKG 12 g, three times a day and calcium carbonate 300 mg, twice a day, orally taken for 6 months; the 23 cases in the positive control group were treated with equal volume of calcium preparation alone and the 52 cases in the blank control group were untreated. All the males were treated in the same way as that in the observed group of female. The BMD level of lumbar vertebrae 2-4 and other related to indexes of bone were determined before and 6 months after treatment. RESULTS: BMD level in the two control groups lowered by 1.8% after 6 months. While in the observed group, BMD lowered slowly or even increased, after 6 months the average BMD increased by 3.1% in females and by 3.9% in males. The effective rate was 91.1% in females and 95.2% in males. After treatment, the levels of tartrate-resistant acid phosphate decreased, estradiol, testosterone and high density lipoprotein increased in the female observed group, while the obove-mentioned indexes unchanged in the control groups. CONCLUSION: GSKG has the effects in increasing sex hormones (both estrogen and androgen), inhibiting bone absorption, slowing down bone mass loss and elevating BMD, it could prevent and treat osteoporosis effectively. Besides, it could increase the level of high density lipoprotein so as to benefit prevention and treatment of cardiovascular diseases. PMID- 12577355 TI - [Effect of zhuhuang frost on wound hydroxy-proline level after anal operation]. AB - OBJECTIVE: To observe the effect of Zhuhuang Frost (ZHF), an external preparation of Chinese herbal medicine, on level of hydroxy-proline (Hyp) in surgical wound after anal operation. METHODS: Fifty postoperational patients after low position simple operation of anal fistula were randomly divided into two groups and treated with ZHF and Mayinglong Musk hemorrhoidal paste (MMHP) respectively. The level of Hyp in granulation of wound was tested using alkaline hydrolysis assay at the 3rd, 7th and 14th day after operation respectively. RESULTS: The Hyp level tested at the 7th and 14th day after operation in the ZHF treated group was higher than that in the MMHP treated group, but those tested at the 3rd postoperational day in the two groups was not different significantly. CONCLUSION: ZHF could increase the Hyp level in postoperational granulation of wound after anal operation. PMID- 12577356 TI - [Effect of Astragalus injection on platelet function and plasma endothelin in patients with early stage diabetic nephropathy]. AB - OBJECTIVE: To study the therapeutic effect of Astragalus injection (AI) in treating early stage diabetic nephropathy (DN) patients. METHODS: The total of 136 early diabetic nephropathy patients were randomly divided into two groups, 50 cases in the conventional treated group and 86 in the AI treated group, the therapeutic course being 3 weeks. Levels of plasma endothelin-1 (ET-1), 24 hrs urinary albumin excretion rate (uAER), and platelet granule membrane protein (GMP 140), 6-keto-prostaglandin F1 alpha(6-keto-PGF1 alpha), and thromboxane B2(TXB2) before and after treatment were determined by radioimmunoassay (RIA) and enzyme linked immunosorbent assay (ELISA) respectively. Moreover, the above-mentioned criteria in 26 healthy subjects were also measured for control. RESULTS: The plasma ET-1, GMP-140, TXB2 and uAER levels in DN patients were higher, but 6-keto PGF1 alpha level was lower than those in healthy subjects. The above elevated criteria in DN patients could be lowered by AI treatment. CONCLUSION: The pathogenesis and development of DN might be closely associated with the changes of plasma ET-1 level and platelet function. Astragalus could improve the above mentioned changes in patients of early stage DN. PMID- 12577357 TI - [Experimental study on effect of xinkang oral liquid on coxsackie B3 viral myocarditis in mice]. AB - OBJECTIVE: To study the therapeutic effect of Xinkang Oral Liquid (XKOL) in treating acute Coxsackie B3 viral myocarditis in mice. METHODS: Viral myocarditis model was established by intraperitoneal inoculation with Coxsackie B3 virus. Mice were randomly divided into 4 groups randomly, the model group, the positive control group (treated with Ribavilin), the XKOL group and the normal control group. Body weight of mice was weighed at the 5th, 10th and 20th day. Weight, macro- and histopathologic changes of heart were observed by microscope and measured by morphometric quantification, and the data were treated statistically. RESULTS: In the XKOL group, the body weight of mice increased, the degree of myocardial necrosis mild and small in size, which has been repaired more completely, the antiviral effect was found in XKOL group. CONCLUSION: XKOL has definite therapeutic effect on viral myocarditis, its effect is better than Ribavilin in protecting myocardium and anti-virus. PMID- 12577358 TI - [Study on effect of nephropathy mixture on genetic expression of liver albumin in rats with adriamycin-induced nephrotic syndrome]. AB - OBJECTIVE: To explore the effect of Nephropathy mixture on genetic expression of liver albunin in rats with adriamycin-induced nephrotic syndrome. METHODS: The rat model of adriamycin-induced nephrotic syndrome was established and the liver albumin mRNA expression level was observed with Northern hybridization and Dot blot quantitative analysis. RESULTS: The liver albumin mRNA expression level in the Nephropathy Mixture treated group was significantly higher than that in the model group and the normal control group (P < 0.05). CONCLUSION: Nephropathy Mixture could up-regulate the liver albumin mRNA expression level and promote the synthesis of albumin in rats with adriamycin-induced nephrotic syndrome. PMID- 12577359 TI - [Effect of reduqing on HL-60 cells in secreting tumor necrosis factor alpha production and on TNF alpha converting enzyme mRNA expression]. AB - OBJECTIVE: To explore the molecular mechanism of inhibiting effects of Reduqing on the release of tumor necrosis factor alpha (TNF alpha). METHODS: Using cytobiologic and molecular biologic technique to observe the effects of Reduqing on HL-60 cells in producing inflammatory cytokine secreting TNF alpha (sTNF alpha) and on mRNA expression of TNF alpha converting enzyme (TACE). RESULTS: (1) Reduqing, diluted in ratio 1:30, could effectively inhibit the increased HL-60 production of sTNF-alpha induced by lipopolysaccharides (LPS); (2) Although no obvious effect on TACE was shown when Reduqing was applied alone on HL-60 cells, there was evident inhibitory effect of Reduqing on TACE mRNA expression enhancement induced by LPS. CONCLUSION: Reduqing could have the double inhibitory effects both on sTNF-alpha production and on the gene expression of its key enzyme, i.e. TACE stimulated by LPS, suggesting that it might be a hopeful and excellent natural TACE inhibitor. PMID- 12577360 TI - [Study on common character of regulative molecular mechanism of Chinese drug bailong and hexamethylen bisacetamide in human cancer cell cycle and their oncogene and tumor suppressor gene expression]. AB - OBJECTIVE: To investigate the common regulative effects of the Chinese drug Bailong and hexamethylen bisacetamide (HMBA) on expressions of oncogenes (c-H-ras and c-myc), and tumor suppressor genes (Rb, p53 and p21) of MGC80-3 in human cancer cell cycle. METHODS: Adopting RNA Northern Blot to survey the levels of gene expressions of MGC80-3 different phases cells treated with Bailong and HMBA respectively. RESULTS: In different phases of MGC80-3 cells treated with Bailong and differentiation inducer HMBA, expressions of oncogenes c-H-ras and c-myc were inhibited by over 50.0%, messenger kinase subspecies PKC-alpha gene is similar with the expression inhibition of oncogenes, except effect of Bailong on the G2 phase in cell cycle. Effect of Bailong differs greatly from HMBA in the expression of tumor suppression genes. The expression of Rb and p21 in cells treated by HMBA did not increase but were inhibited by 39.5% and 33.3% respectively in G1 phase. The level of Rb gene expression was decreased, too by 3.0% in S phase. Comparison with HMBA the expression of Rb and p21 genes were increased after treatment by Bailong in all cell cycle. But the effect of Bailong on the expression of p53 gene which was increased obviously by 125.0%-233.4% in majority phase of MGC80-3 cells is similar to HMBA. CONCLUSION: (1) The effect of Bailong on the regulation of oncogenes and tumor suppressor gene is similar to HMBA but the effect of Bailong is better than that of HMBA. (2) Molecular mechanism of the Bailong or HMBA on the proliferative inhibition and differentiation of MGC80-3 related to regulation of the Bailong and HMBA on the oncogenes and tumor suppressor genes in cell cycle of MGC80-3. PMID- 12577361 TI - [Experimental study on oncogenicity of Aristolochia manshuriensis in rats]. AB - OBJECTIVE: To observe the oncogenetic process, biological behavior, pathological and immunohistochemical features of tumor induced by Aristolochia manshuriensis (AM) in rats. METHODS: Acute renal injury model was established with AM docoction in different dosages by gastrogavage to observe the histomorphologic and immunohistochemical features dynamically. RESULTS: (1) At month 0, 1 and 3, the occurrence of renal tumor or tumor-like proliferation was not observed; (2) At month 6, the occurrence of renal tumor-like proliferation in all the three AM dosage groups (50 g/kg, 30 g/kg and 20 g/kg) was 100.0%. Immunohistochemical examination conducted in 2 rats showed that the short spindle-shaped interstitial cells were expressed positively both by vimentin and proliferative cell nuclear antigen (PCNA), but were shown negative for smooth muscle actin (SMA) and p53; (3) At month 6, the occurrence of renal tumor in the three dosage groups was 42.8%, 25.0% and 0% respectively, including 4 cases of renal mesenchymal tumor and 1 case of nephroblastoma. Immunohistochemical examination conducted in 3 cases of renal mesenchymal tumor showed that the short spindle-shaped tumor cells expressed both by vimentin and PCNA, and SMA and p53 were positive for well differentiated tumor cells. (4) The occurrence of extrarenal tumor in the three dosage groups was 14.3%, 12.5% and 12.5% respectively, 1 case of mammary duct epithelial tumor, 1 thyroid follicle epithelial tumor and 1 skin appendicular epithelial tumor. No tumor occurred in the control group. CONCLUSION: Large dosage of AM is oncogenic. The occurrence of renal tumor was relatively high, and the histological type is mainly mesenchymal. Vimentin, SMA, PCNA and p53 positive expression was shown for well-differentiated renal mesenchymal tumor. The occurrence of extrarenal tumor is rather low. PMID- 12577362 TI - [Progress in research on PC-SPES anti-cancer agents in prostatic carcinoma in USA]. PMID- 12577365 TI - [Discussion on the modernization of Chinese herbal medicine]. PMID- 12577364 TI - [Pharmacological action of Chinese herbal medicine for antagonizing diabetes mellitus]. PMID- 12577366 TI - [Experience on treatment of pancreatitis by integrated traditional and Western medicine]. PMID- 12577367 TI - [Effect of combination of Chinese herbal drugs on clinical bioavailability of ferulic acid in serum]. AB - OBJECTIVE: To observe the effect of combination of Chinese herbal drugs on bioavailability of Ligusticum walichii (LW) derived ferulic acid (FA) in serum. METHODS: Healthy volunteers were given LW decoction (1 g/kg), LW combined with Paeonia veitchii (PV, 1:1) decoction (2 g/kg) and Coronary Heart No. 2 (CH-II, 3 g/kg, containing LW 0.5 g/kg) once respectively. The serum FA concentration-time (c-t) curve was performed by HPLC using the method established by the authors, and the area under that curve (AUC) was calculated and compared. RESULTS: The FA c-t curve in the three groups was double peak curve. As compared with the AUC in subjects who were administered LW decoction, the AUC in those taken LW + PV decoction was lower [(15734.88 +/- 7737.97) ng/(min.ml) vs (5717.92 +/- 6124.63) ng/(min.ml)], P < 0.05), while that in those taken CH-II was relatively higher [(8072.00 +/- 4424.31) ng/(min.ml), P > 0.05)]. CONCLUSION: Combination of LW and PV could significantly lower the bioavailability of FA, but when LW was added in CH-II, the FA bioavailability was not affected. PMID- 12577368 TI - [Clinical and experimental study on multiple sclerosis with bushen gusui tablet]. AB - OBJECTIVE: To observe the therapeutic effect of Bushen Gusui tablet (BSGS) in treating multiple sclerosis (MS) and its effects on experimental allergic encephalomyelitis (EAE) in guinea pigs. METHODS: Forty-three MS patients were treated with BSGS and their clinical symptoms, signs of nerve function, recurrent frequency, evoked potential and changes in magnetic resonance imaging (MRI) were observed. The EAE model of guinea pigs was induced by homogenate of rabbit spinal cord and complete Freund's adjuvant (CFA), the animals were treated with BSGS and compared with prednisone acetate, which was served as control. The mortality and pathomorphology of EAE animals were observed. The contents of serum interleukin-2 (IL-2), interleukin-6 (IL-6), tumor necrosis factor (TNF) as well as myelin basic protein (MBP) were determined. RESULTS: BSGS could improve symptoms and signs of MS patients and reduce recurrent frequency. The total effective rate was 88.37%. High dose BSGS could obviously reduce incidence of EAE, inhibit inflammatory reaction of brain and spinal cord as well as demyelination, and simultaneously inhibit the activity of serum IL-2, IL-6, TNF and MBP, in comparing with model group (P < 0.01). There were insignificant difference as compared with prednisone acetate group (P > 0.05). CONCLUSION: BSGS had certain effect on both MS patients and EAE model animals, which indicated that it was worth further studying and clinical application. PMID- 12577369 TI - [Assessment on effect of treatment of chronic plaque type psoriasis by combination therapy of composite shendi decoction and diyin tablet]. AB - OBJECTIVE: To seek for a better therapy for chronic plaque type psoriasis. METHODS: The effect of 35 patients in the tested group treated with combination therapy of composite Shendi Decoction (CSDD) and Diyin Tablet (DYT) was observed and compared with the effect of those treated by CSDD alone (25 cases) or by DYT alone (18 cases) as the control groups. RESULTS: The total effective rate in the treated group was 94.29%, and that in the two control groups was 88.00% and 83.33% respectively. The markedly effective rate in the treated group was 62.86%, and that for the control group was 36.00% and 27.78% respectively. Ridit test showed significant difference among the effectiveness in the three groups, while chi 2 showed the difference of the total effective rate in the three groups was insignificant, but that of the markedly effective rate was significant. Serum IL 8 and plasma endothelin levels of 11 cases in the treated group were examined before and after treatment, the results showed that both the two parameters were reduced after the combination therapy. CONCLUSION: The effect of combination therapy of CSDD and DYT was better than that of CSDD or DYT alone, therefore, it is worth further studying and spreading. PMID- 12577370 TI - [Clinical and experimental study on treatment of childhood hyperkinetic syndrome with yizhidan]. AB - OBJECTIVE: To observe the clinical effect of Yizhidan (YZD) in treating childhood hyperkinetic syndrome (CHS) and its mechanism. METHODS: Random single blind method was used in clinical study to observe the clinical effect of YZD and compared with that of Ritalin. The dysmnesia model animal was used to observe the effect of YZD on learning memory function and cerebral monoamine neurotransmitter content. RESULTS: The markedly effective rate and the total effective rate of YZD were 45.56% and 87.78%, which were similar to those of Ritalin (53.33% and 86.67%). The Conners index lowered in both groups, showing insignificant difference (P > 0.05). Experimental observation showed that YZD could significantly improve the soft neurologic signs and abnormal encephalogram with less side-effects in comparing with Ritalin (P < 0.05). Moreover, YZD could obviously improve the learning memory disturbance induced by scopolamine, sodium nitrite and ethanol, and significantly increased the contents of DA in cerebral tissue, as compared with that of the control group, the difference was significant (P < 0.01). It also showed effect on DOPAC, NE, HVA, 5-HT and 5-HIAA of mice in certain degree. CONCLUSION: YZD is an effective drug in treating CHS, its mechanism might be probably related to the improvement of the memory and the enhancement of DA receptor in cerebral tissue. PMID- 12577371 TI - [Effect of Tripterygium wilfordii on Th1, Th2 cytokines production in asthma patients]. AB - OBJECTIVE: To observe the effect of Tripterygium wilfordii on Th1, Th2 cytokines in asthma patients for further study on the therapeutic mechanism. METHODS: Twelve patients of middle or severe asthma were treated by Tripterygium polyglucoside 40 mg or 60 mg daily for 4 weeks. Blood of patients was colleted before and after treatment for serum and peripherol blood mononuclear cells (PBMC) preparation. The prepared PBMCs were stimulted in vitro with Concanavalin A (ConA) for 6 hrs and followed by culturing with Triptolide for 24 hrs and then the supernatant was collected. The concentration of interleukin-2(IL-2), -4(IL 4), -5(IL-5) and interferon-gamma(IFN-gamma) in serum and in the supernatant were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Serum levels of IL-2, IL-4 and IL-5 of patients decreased significantly after treatment of Tripterygium polyglucoside (P < 0.01), but IFN-gamma level was under the detection sensitivity both before and after treatment. Triptolide could inhibit PBMC to secrete IL-2, IL-4 and IL-5 in vitro (P < 0.01), but IFN-gamma was also under the detection sensitivity. CONCLUSION: The marked inhibition of Th2 cytokine expression by Tripterygium was the important mechanism of it in treating asthma. But the fact that Tripterygium also showed inhibition on Th1 cytokine indicated that the inhibition of Tripterygium on Th2 and Th1 cytokines was non specific. PMID- 12577372 TI - [Effect of tripterygium polyglucoside on T-lymphocyte subsets and serum interleukin-5 level in asthma patients]. AB - OBJECTIVE: To investigate the therapeutic mechanism of Tripterygium polyglucoside (TII) in treating asthma patients. METHODS: Thirty asthma patients of middle or sever leoel were selected and randomly divided into three groups, Group A treated with TII, Group B treated with prednisone and Group C, the control group. Their peripheral CD4+, CD8+ T-lymphocytes were counted by flow cytometer and serum interleukin-5 (IL-5) level determined by ELISA before and after 4 weeks' treatment. RESULTS: In Group A, after treatment, CD4+ T-lymphocytes reduced from 0.462 +/- 0.035 to 0.426 +/- 0.039 (P < 0.01), CD8+ increased from 0.201 +/- 0.045 to 0.253 +/- 0.043 (P < 0.01), and serum IL-5 median concentration reduced from 65.3 ng/L to 10.9 ng/L (P < 0.01). Similar results was revealed in Group B, while there was insignificant change of the parameters in Group C. The serum concentration of IL-5 was positively correlated with the peripheral CD4+ count (r = 0.61, P < 0.01). CONCLUSION: TII is highly effective in treating asthma through regulating T-lymphocyte subset disorder and inhibiting production of IL-5. PMID- 12577373 TI - [Therapeutic mechanism of combination therapy of ligustrazine and nitrendipine in treating patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To study the therapeutic mechanism of combination therapy of ligustrazine and nitrendipine in treating patients with chronic obstructive pulmonary disease (COPD). METHODS: Thirty COPD patients divided in to 3 groups (10 in each) were treated with ligustrazine, nitrendipine and ligustrazine plus nitrendipine respectively, and the changes of hemorrheologic parameters, plasma endothelin (ET-1), thromboxane A2(TXA2) and platelet-P-selectin (CD62P) before and after treatment were observed. RESULTS: The combination therapy of ligustrazine and nitrendipine could lower the levels of plasma ET-1, TXA2, CD62P and the hemorrheologic parameters. CONCLUSION: Combination of ligustrazine and nitrendipine showed a therapeutic effect better than that of the two drugs used separately. Its effect in lowering pulmonary circulation resistance is related with the lowering of plasma vaso-contrictive factor and the changing of hemorrheologic properties. The integrated traditional Chinese and western medical therapy is valuable in treating COPD. PMID- 12577374 TI - [Effect of Puerarin on platelet activating factors CD63 and CD62P, plasminogen activator inhibitor and C-reactive protein in patients with unstable angia pectoris]. AB - OBJECTIVE: To explore the action of platelet surface activity protein (platelet granule membrane protein CD63 and lysosome membrane protein CD62P), plasminogen activator inhibitor (PAI-1) and C-reactive protein (C-RP) in occurrence and development of unstable angina pectoris (UAP), and the effect of Puerarin on them. METHODS: Patients with UAP were randomly divided into the treated group (32 cases, treated with Puerarin) and the control group (27 cases, treated with Ticlid), the therapeutic course was 4 weeks for both groups. Changes of CD63, CD62P, PAI-1 and C-RP before and after treatment were observed. RESULTS: The levels of CD63, CD62P, PAI-1 and C-RP were higher in UAP patients than those in normal subjects and in the patients with stable angina pectoris (P < 0.05 or P < 0.01). These parameters were increased along with the aggravating of patients in Braunwald's degree. After 4-week treatment, the above-mentioned parameters lowered in both groups (P < 0.05 or P < 0.01), and comparison between the two groups showed no significant difference statistically. CONCLUSION: Platelet activation, plasminogen activator and C-RP play important roles in the occurrence and development of UAP. The obvious effect of Puerarin in anti-platelet activation, improving plasminogen activator and relieving inflammatory reaction was of great importance in preventing the occurrence and development of acute coronary syndrome clinically. PMID- 12577375 TI - [Clinical observation on treatment of ulcerative colitis by soothing liver and activating spleen]. AB - OBJECTIVE: To observe the effect of soothing Liver and activating Spleen method (SLAS) of TCM in treating ulcerative colitis (UC). METHODS: Patients were randomly divided into 2 groups, the 33 cases in the treated group were treated with Ningxie Powder (NXP) and the 30 cases in the control group were treated with salicylazosulfapyridine (SASP). The improvement in clinical symptoms scoring, therapeutic effect in various treatment courses, changes in immune indexes as well as the condition in 1-year follow-up were observed. RESULTS: The initial therapeutic effect after 3 months treatment showed insignificant difference between the two groups. However, the effect in the treated group was better than that in the control group both short-term (after 9 months treatment) and long term (for 1-year follow-up study) (P < 0.05). CONCLUSION: NXP, a Chinese herbal drug for soothing Liver and activating Spleen, shows good and persistent effect in treating ulcerative colitis and has favorable prospect in application. PMID- 12577376 TI - [Experimental study of inhibitory effect of kangxian recipe on TGF-beta 1 induced hepatocyte apoptosis]. AB - OBJECTIVE: To study the inhibitory effect of Kangxian Recipe (KXR) on TGF-beta 1 induced hepatocyte apoptosis. METHODS: The in vitro model of hepatocyte apoptosis was established by cell biologic methods, utilizing the characteristics of TGF beta 1 to observe the inhibitory effect of KXR on hepatocyte apoptosis. RESULTS: TGF-beta 1 induced apoptosis of hepatocyte in a dose-dependent manner. The apoptosis rate of 2.2.15 cells was 63% when 500 ng/L TGF-beta 1 was used, while for HepG2 cell, it was merely 44%. After treatment of 20 ng/L KXR, the apoptosis rate of the two kinds of cell lines lowered to 33% and 24% respectively. The inhibition rate of both groups was about 50%. CONCLUSION: KXR had strong inhibitory effect on hepatocyte apoptosis induced by TGF-beta 1. PMID- 12577377 TI - [Effect of ginsenosides in inducing proliferation and transcription factor of erythrocytic, granulo-monocytic and megakarocytic cell lines]. AB - OBJECTIVE: To investigate the action of Ginsenosides (GS) in inducing transcription factor c-fos and GATA-1 to explore the mechanism of GS in hematopoietic cells. METHODS: The proliferation effects of GS on granulocytic (HL 60), monocytic (U937), erythrocytic (K562) and megaryocytic (Meg-01) cell lines were observed by using proliferation test of MTT and colony formation of progenitor cells. The combining reaction of transcription factors c-fos and GATA 1 with nuclear protein antigen were analyzed by Western Blot after being treated by GS. RESULTS: (1) GS (10 micrograms/ml) could stimulate and promote proliferation of 3 cell lines with significant difference between GS and non-GS control (P < 0.05 in all) in both MTT test and colony assay. (2) After treatment with GS, c-fos protein in HL-60, K562 and Meg-01 cell lines was increased by 1.5, 2.0 and 2.5 fold respectively, while U937 cell did not express c-fos. (3) Except that U937 cell hadn't expressed GATA-1, the other cell lines after the treatment by GS, the GATA-1 protein level was elevated to 1.5, 2.1 and 1.3 fold of that before treatment. CONCLUSION: The proliferation of three lines initiated by GS was involved in transcription factor c-fos or GATA-1, which could pay the role in the GS induced up-regulation correlated with proliferation and differentiation of hematopoiesis. PMID- 12577378 TI - [Therapeutic effect of Astragalus and Angelica on renal injury induced by ischemia/reperfusion in rats]. AB - OBJECTIVE: To observe the effect of Astragalus and Angelica on acute renal injury. METHODS: Using the ischemia/reperfusion model established by blocking blood flow through clamping of bilateral renal artery for 45 min, the changes of glomerular filtration rate (GFR), renal plasma flow (RPF), fractional excretion of sodium (FENa), urinary volume (UV) and mean arterial pressure (MAP) as well as the morphological change of kidney before and after ischemia/reperfusion were observed. RESULTS: Astragalus and Angelica could promote recovery of RPF and GFR after ischemia/reperfusion, prevent the oliguria or shorten the oliguria period, reduce the increment of FENa and improve the histomorphological injury of kidney. CONCLUSION: Astragalus and Angelica have certain effect in protecting kidney from acute renal injury. PMID- 12577379 TI - [Experimental study of protective effect of shenmai injection on endotoxin induced systemic inflammatory reaction syndrome and multiple organ dysfunction syndrome]. AB - OBJECTIVE: To study the protective effect of Shenmai injection (SMI) on systemic inflammatory reaction syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) in animal model of SIRS, MODS induced by lipopolysaccharide (LPS). METHODS: Male BALB/c mice, 5-7 weeks old were randomly divided into 3 groups, the normal control group, the LPS model group and the LPS + SMI group. The rectal temperature, peripheral white blood cell count (WBC), blood biochemical examination, histopathologic changes of lung and liver as well as the blood levels of endotoxin and tumor necrosis factor alpha (TNF alpha) were determined before and after treatment. RESULTS: LPS could induce endotoxemia and high serum level of TNF alpha, decrease the rectal temperature and WBC, reduce blood glucose and increase serum triglyceride and cholesterol levels. Pathological examination showed that LPS could cause pulmonary alveolar congestion, edema, exudation, capillary dilation and inflammatory cell infiltration in liver and lung tissue. SMI could significantly raise the low body temperature caused by LPS and reduction of WBC, improve the hypoglycemia and high plasma TNF alpha level, alleviate the pathologic changes in organs and reduce the plasma level of LPS (endotoxin). CONCLUSION: SMI has marked effect in protecting LPS caused SIRS, MODS, the mechanism might be related with the lowering of LPS (endotoxin) level and reducing of TNF alpha secretion. PMID- 12577380 TI - [Experimental study of protective effect of tongxinluo capsule on isoproterenol induced myocardial injury in rats]. AB - OBJECTIVE: To study the protective effect of Tongxinluo capsule (TXLC) on myocardial injury induced by isoproternol. METHODS: Myocardial injury was induced in 34 rats by subcutaneous injection of isoproterenol (85 mg/kg). The experimental animals were randomly divided into the control group, isoproterenol group and TXLC group. The histopathological change of myocardia was investigated by HE staining, the myocardial cell apoptosis were observed by TUNEL method and the ultrastructure of myocardial cell examined by electron microscope. RESULTS: No cell degeneration and necrosis, only very few cells of apoptosis positive were found in the control group. While in the isoproterenol group, marked necrosis of myocardial tissue and increase of apoptosis cells were found, and characteristic changes of cell apoptosis were observed under electron microscope. After TXLC treatment, the myocardial necrosis and cell apoptosis were markedly alleviated. CONCLUSION: Isoproterenol could induce myocardial necrosis and apoptosis, TXLC could alleviate the myocardial injury through preventing myocardial cell necrosis and apoptosis. PMID- 12577381 TI - [Effect and mechanism study of kidney tonifying and blood circulation activating Chinese drugs on development of mammary glands in female juvenile rats]. AB - OBJECTIVE: To study the effect and mechanism of Kidney tonifying and blood circulation activating Chinese drugs (KB) on development of mammary gland. METHODS: Using the female juvenile rats as the experimental targets, the level of serum hormone, the amount and affinity of estradiol receptor (ER) and progesterone receptor (PR) as well as the DNA content and relative weight of the rats' mammary glands were measured to study the effect of KB at the pre-receptor, receptor and post-receptor level. RESULTS: KB markedly enhanced the serum level of estradiol, progesterone and the growth hormone (GH) at pre-receptor level, increased the number and affinity of estradiol and progesterone receptors both in the cytoplasm and nucleus at the receptor level. While at the level of post receptor, it increased the DNA content and the weight index of mammary glands. CONCLUSION: The Kidney tonifying and blood circulation activating Chinese drugs are effective in promoting the growth and development of mammary glands. PMID- 12577382 TI - [Current status of injury caused by Chinese herbal medicine and its counter measure]. PMID- 12577383 TI - [Status of and prospect on experimental studies of single Chinese herb inducing tumor cell apoptosis]. PMID- 12577384 TI - [Summary of the International Academic Conference of integrated Traditional Chinese and Western Medicine on Health Preservation and Rehabilitation]. PMID- 12577385 TI - [Development and creation--to elevate the level of manufacturing nutritious food]. PMID- 12577386 TI - [Study on diagnosis and treatment of infections multiple organ dysfunction syndrome by integrated traditional Chinese and Western medicine]. AB - OBJECTIVE: To explore the diagnostic and therapeutic approach of integrated traditional Chinese and western medicine (TCM-WM) on infectious multiple organ dysfunction syndrome/multiple system and organ failure (MODS/MSOF) for elevating the successful rate of rescuing the patients. METHODS: Diagnosis with western medicine and Syndrome Differentiation of TCM in 225 in-patients of acute infectious disease complicated with MODS/MSOF were conducted, and TCM treatment, based on western medical comprehensive treatment, was given to observe the effect and explore the mechanism of the TCM-WM therapy. RESULTS: Up to the end of 1998, 161 cases of the 225 cases were successfully cured and 64 died, the mortality being 28.4%. Among them, 58 out of 140 cases of MSOF died, the mortality was accounted for 41.4%. In 106 cases conformed to the diagnostic criteria of MSOF proposed by Professor Knaus WA, USA, 52 cases were cured successfully and 54 died, the mortality being 50.9%. CONCLUSION: TCM-WM treatment could elevate the therapeutic effect in treating MODS, the mechanism might be through improving the hemodynamic and hemorrheologic condition of patients to relieve nail-fold microcirculation disorder; influencing the levels of cytokine and inflammatory mediator, so as to alleviate the systemic inflammatory reaction, it might also abate the inhibited condition of gastro-intestinal motility, alleviate the intestinal flora imbalance, prevent intestinal bacteria and endotoxin malposition, and protect cells from peroxidation. PMID- 12577387 TI - [Twelve cases of malignant hematopathy treated by combined therapy of hematopoietic stem cell transplantation and Chinese herbal medicine]. AB - OBJECTIVE: To evaluate the effect of hematopoietic stem cell transplantation combined with Chinese herbal medicine in treating malignant hematopathy. METHODS: Allo-bone marrow transplantation (Allo-BMT) or alloperipheral blood stem cell transplantation (Allo-PBSCT), with conditioning regimen of TBI + Cy or Bu + Cy, was used to treat 4 cases of chronic granulocytic leukemia (CGL, 3 of chronic phase and 1 of acceleration phase) and one case of acute non-lymphocytic leukemia (ANLL). And auto-BMT or auto-PBSCT, with conditioning regimen of MAC or MAC + VP16, was used to treat 7 cases of hematopathy, including 5 cases of ANLL (3 of CR1 and 2 of CR2) and 2 cases of malignant lymphoma (1 of first occurring and 1 of relapse). Chinese herbal medicine was given orally to all the 12 patients after' transplantation according to TCM Syndrome Differentiation. RESULTS: All patients were successfully engrafted. Within the median follow-up period of 18 (4 70) months, one patient (8.3%) died of transplantation-related death, 3(25.0%) relapsed and 8 (66.7%) survived free of original disease. The transplantation effective rate was 88.8% as counting by ANLL CR1 and CGL chronic phase. CONCLUSION: Auto-BMT or auto-PBSCT in CR1 stage of acute leukemia could reduce the relapse rate, when there was no matched bone marrow donor; allo-BMT or allo PBSCT in chronic stage could result in long-term disease-free survival of patients; Chinese herbal medicine administration in patients of malignant hematopathy might reduce the complications and plays certain role in promoting recovery of hematopoietic function. PMID- 12577389 TI - [Clinical study on treatment of exogenous infection caused fever by infantile umbilical paste]. AB - OBJECTIVE: To observe the effect of infantile umbilical paste (IUP) in abating infantile fever. METHODS: Two hundred and twenty cases were randomly divided into two groups, the 120 cases in Group A were treated with IUP and the 100 cases in Group B treated with compound Aspirin (APC). The effect in abating fever and side effects were observed. RESULTS: The total effective rate of the two groups were not statistically different (P > 0.05). But a better effect in abating fever was shown in Group A 4 hrs later (P < 0.01). The effect in patients of age under 1 year was the best. CONCLUSION: IUP has good effect in abating infantile fever, and it shows no influence on white blood cells, liver and kidney functions. PMID- 12577388 TI - [Clinical and experimental study on the ovulation promoting effect of yangjing decoction]. AB - OBJECTIVE: To observe the effect of Yangjing Decoction (YJD) to promote ovulation and to explore its mechanism. METHODS: The clinical effect of YJD in treating patients of functional non-ovulation and changes of gonadotropic hormone, sex hormone and diameter of main follicle before and after treatment were observed. In experimental study, the influence of YJD on monoamine neurotransmitter of hypothalamus, endorphin (beta-EOP), serum sex hormone, trace element, endometrium and histomorphology of ovary were observed. RESULTS: In the 36 patients observed clinically, the ovulation rate was 61.1%, the serum contents of follicular stimulation hormone (FSH), luteinizing hormone (LH) and estradiol (E2) increased and the diameter of main follicle enlarged significantly after treatment (P < 0.01, P < 0.001). The experimental study showed that the totals of follicle at different levels, the corpus luteum and the proportion of superior follicle in ovary were improved obviously in the YJD group as compared with those in the control group. After YJD treatment the hypothalamus contents of monoamine neurotransmitter, including norepinephrine (NE) and dopamine (DA) increased, the 5-hydroxytryptamine (5-HT) 5-hydroxyindoleaceticacid (5-HIAA) and beta-EOP decreased significantly, and the NE/DA ratio and NE/5-HT ratio elevated obviously (P < 0.05, P < 0.001). The serum levels of E2, P and trace elements, including Zn, Cu and Fe increased significantly (P < 0.05, P < 0.01). CONCLUSION: YJD is an effective recipe in promoting ovulation, the effect may be achieved through the multi-linked, multi-systemetic synthetic effect of hypothalamus, hypophysis and ovary, etc. PMID- 12577390 TI - [Study for effect of kangyanling on cytokine and C-reactive protein in patients of systemic inflammatory reaction syndrome and multi-organ dysfunction syndrome after abdominal surgery]. AB - OBJECTIVE: To observe the clinical effect and mechanism of Kangyanling (KYL) in treating patients with systemic inflammatory reaction syndrome and multi-organ dysfunction syndrome (SIRS/MODS) after abdominal surgery. METHODS: Eighty-two patients of SIRS/MODS after abdominal surgery were divided into two groups according to admission time, the KYL treated group (n = 35) and the control group (n = 47). The levels of serum C-reactive protein (CRP), plasma tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) were measured at the 1st, 3rd and 7th day post-operationally. RESULTS: The levels of CRP, TNF-alpha and IL-6 decreased gradually after surgical operation in both groups, but the reducing velocity was shorter in the KYL group than that in the control group, so the comparison of the levels in the two group showed significant difference on the 3rd day after operation (P < 0.05). CONCLUSION: KYL could inhibit the release of inflammatory mediator and relieve the inflammatory response so as to treat post operational SIRS/MODS effectively. PMID- 12577391 TI - [Effect of Acanthopanax senticosus injection on plasma and urinary endothelin in early stage of diabetic nephropathy]. AB - OBJECTIVE: To explore the mechanism of Acanthopanax senticosus (AS) on diabetic nephropathy (DN) patients by observing its effect on urinary albumin excretion (UAE) and renal endothelin (ET) metabolism. METHODS: Forty-seven patients of diabetes mellitus (DM) type 2 with persistent microalbuminuria and normotension were randomly divided into two groups, the conventional treatment group (control group) and the AS treatment group. The treatment course was 8 weeks. The changes of UAE, plasma and urinary ET levels before and after treatment were observed. RESULTS: Levels of UAE, plasma and urinary ET lowered significantly (P < 0.01) in the AS groups, while no significant change was shown in the control group. By correlation analysis, the decrement of UAE was positively correlated with the decrement of urinary and plasma ET levels (r = 0.72, P < 0.01 and r = 0.63, P < 0.01). CONCLUSION: The protective effect of AS injection on DN is probably correlated with its inhibition on ET synthesis in kidney. PMID- 12577392 TI - [Study on effect of shenmai injection in protecting myocardium against ischemia reperfusion injury in thrombolytic therapy with urokinase for acute myocardial infarction patients evaluated by 99mTc-MIBI myocardial imaging]. AB - OBJECTIVE: To evaluate the myocardial protecting effect of Shenmai injection (SMI) against ischemia/reperfusion injury in thrombolytic therapy with urokinase (UK) for acute myocardial infarction patients by 99mTc-MIBI myocardial imaging (SPECT). METHODS: Five hundred and thirty-seven patients were randomly divided into two groups. The SMI group (n = 292) was treated with thrombolytic treatment plus SMI and the control group (n = 245) with thrombolytic treatment only. Single photon emission computerized tomography (SPECT) was carried out on the 7th day after thrombolysis to determine the ischemic myocardial area (IMA) and ejection fraction (EF) in both groups and compared. RESULTS: The reperfusion rate of infarction related area (IRA) in the two groups was not different significantly (72.26% vs 72.65%, P > 0.05). The IMA in patients of the SMI group, no matter with or without reperfused IRA (211 cases and 81 cases) respectively, was significantly lower than that in the control group (178 cases and 67 cases) respectively, P < 0.01 and P < 0.05 respectively. The EF value in the SMI group was significantly higher than that in the control group (P < 0.01). CONCLUSION: Using SMI in early stage of thrombolytic treatment in acute myocardial infarction could significantly reduce IMA and increase EF. SMI showed good protective effect against myocardial ischemia/reperfusion injury in thrombolytic treatment. PMID- 12577393 TI - [Clinical and experimental study of effect of yangxin fumai oral liquid in treating patients with extrasystole]. AB - OBJECTIVE: To evaluate the therapeutic effect of Yangxin Fumai Oral Liquid (YFOL), a Chinese herbal medicine for nourishing heart and restoring pulse, in treating patients with extrasystole. METHODS: The effect of YFOL was observed in treating 30 patients with different kinds of extrasystole and compared with that in 30 patients treated by propafenone. The effect of YFOL on experimental arrhythmia was studied in animals as well. RESULTS: Clinical observation showed that the effect of YFOL against extrasystole in the two groups was similar, but the YFOL group showed better effect in symptom improvement (P < 0.01) with no marked side-effects. Experimental study showed that YFOL could reduce the chloroform induced ventricular fibrillation occurrence in mice, delay the initiating time of ventricular extrasystole, tachycardia and fibrillation induced by aconitine, BaCl2 and coronary artery ligation in rats, or shorten the lasting time of arrhythmia, reduce the attacking rate of ventricular extrasystole. There was significant difference in comparing with the control group (P < 0.05, P < 0.01). CONCLUSION: YFOL is a good and convenient Chinese herbal preparation for different kinds of extrasystole with low toxic and side-effects in clinical practice. PMID- 12577394 TI - [Changes of cerebral beta-endorphin in rats treated with combination therapy of melatonin and electroacupuncture]. AB - OBJECTIVE: To explore the strengthening of acupuncture analgesic mechanism on the level of beta-endorphin and proopimelanocortin mRNA expression in the arcuate nucleus of hypothalamus in rats following electroacupuncture (EA) combined with melatonin (MEL). METHODS: Integrated optical density (IOD) was measured by ABC immuno-histochemical and in situ hybridization technique with computerized image processing. The rat's brain was coronally sectioned after combination of EA and MEL. RESULTS: IOD of beta-endorphin-like immunopositive substance in rat's brain was lowered significantly, which was measured after MEL (60 mg/kg) was injected intraperitoneally and followed by EA 30 min later for 30 min, and the IOD of cerebral POMC mRNA positive substance increased significantly 10 hrs later. CONCLUSION: The mechanism of MEL in enhancing EA analgesic effect might be related with the release and synthesis of beta-endorphin. PMID- 12577395 TI - [Study on immunomodulating mechanism of the active fraction of liuwei dihuang decoction]. AB - OBJECTIVE: To investigate the immunomodulating mechanism of 3A, an active fraction extracted from the ethanol-soluble portion of Liuwei Dihuang Decoction (LWDHD). METHODS: Influence of 3A on T-lymphocyte subsets, IgG production of B cells, interferon-gamma (IFN-gamma) mRNA expression in cyclophosphamide (Cy) treated mice, tumor bearing mice and senescence accelerated mouse-prone/8 (SAMP8) were assessed in vivo and in vitro. RESULTS: Oral administration of 3A could elevate the decreased T-lymphocyte subsets in all model mice, it also could promote the IgG secretion of splenic cells in SAMP8 and increase IFN-gamma mRNA expression in Cy-treated mice. CONCLUSION: The immunomodulating effect of 3A is related with its effects in regulating ratio of T and B cells or in improving their functions. PMID- 12577396 TI - [Effect of fuzheng kangbai granule on immune function and survival time in minimal residual leukemia model in mice]. AB - OBJECTIVE: To observe the effects of Fuzheng Kangbai Granule (FZKBG) on immune function and survival time in minimal residual leukemia (MRL) model mice and study its mechanism. METHODS: MRL model mice were established by hypodermic inoculation with L7212 cells following intraperitoneal injection of cytoxan (CTX) 25 mg/kg 3 days later, and divided into the control group and FZKBG group. The changes of T-lymphocyte subsets, including CD3+, CD4+ and CD8+, and the survival time in model mice were observed. RESULTS: Comparing with the control group, FZKBG could obviously increase both the percentage and absolute value of CD3+ and CD4+ lymphocytes and prolong the survival time of model mice, the prolongation rate being 29.6%-60.4%. CONCLUSION: FZKBG could markedly prolong the survival time of MRL mice, its mechanism might be through elevating the immune function and inhibiting the leukemic cells in model mice. PMID- 12577397 TI - [Experimental study on preventive and therapeutic effect of bushen huoxue recipe on autoimmune premature ovarian failure model]. AB - OBJECTIVE: To investigate the mechanisms of Bushen Huoxue Recipe (BSHXR), a Chinese herbal medicinal preparation for tonifying Kidney and invigorating blood circulation, to prevent and treat autoimmune premature ovarian failure (POF) model. METHODS: Female 8-10 weeks old BAL B/c mice were immunized by intracutaneously injecting porcine ovum zona pellucida (ZP), isolated by immuno chromatography, in multiple points of two hind footpads to establish the POF model and treated with BSHXR started from early stage of immunization (prevented group) or after 3 times of injection (treated group). Changes in vaginal smears, serum estradiol (E2), antibody titer against ZP, response of splenic lymphocyte to ZP stimulation of different concentrations, and numbers of ovarian follicles and corpus luteum were analyzed. RESULTS: Serum E2 level in the prevented and treated mice was all higher than that in the non-treated immunized model mice (the control group), P < 0.01 and P < 0.05 respectively. But the anti-ZP titer lowered significantly after BSHXR administration, as compared with that in the control group. Level of antibodies in the treated group was lower than that of the control, and it was also lower in the prevented group than that in the control. The histo-morphological examination showed that the developing follicles and corpus luteum after BSHXR medication in both prevented and treated group increased significantly as compared with that of the control group (P < 0.05 and P < 0.01 respectively). Splenic lymphocyte in the immunized model showed a higher antigen-specific proliferation reaction than that in non-immunized animal, and the reaction was ameliorated by BSHXR medication. CONCLUSION: BSHXR could recover part of the ovarian function in POF mice mainly through inhibiting specific immune injury to revive the remnant follicles in ovary. The preventive effect of BSHXR was superior to the therapeutic effect of it. PMID- 12577398 TI - [Effect of the tetrandrine on morphine induced hyperactivity and reinforcement in mice]. AB - OBJECTIVE: After single administration of morphine and the motion activity was measured by ambulometer, conditioned place-preference paradigm was used to study the reinforcing effect of morphine, climbing behavior was used to evaluate the relation with Dopaminergic system and immediate early expression of c-fos gene was in brain was showed by immunohistochemical method. RESULTS: Single administration of morphine could induce hyperactivity, repeated treatment would produce a conditioned place-preference response, tetrandrine 30 or 60 mg/kg hypodermic injection could inhibit the morphine induced hyperactivity, 60 mg/kg could inhibit the conditioned place-preference response but no influence on climbing behavior in mice was found. Tetrandrine could inhibit the c-fos gene expression in nucleus accumbens, ventral tegmental and prefrontal cortex in place preference model formed by morphine. CONCLUSION: Tetrandrine could inhibit the hyperactivity and conditioned place preference response induced by morphine, it might relate to reducing the c-fos gene expression in special area of brain in mice. PMID- 12577399 TI - [Integration of traditional Chinese and Western medicine]. PMID- 12577400 TI - [Analysis on articles dealing with "Treatise on febrile diseases" and its author published in CJIM from starting of the journal]. PMID- 12577401 TI - [Survey of researches on anti-pilatelet effects of saponins contained in Chinese herbal medicine]. PMID- 12577402 TI - [Progression of integrated traditional Chinese and Western medical researches on diabetic heart diseases]. PMID- 12577404 TI - [The first heart-lung transplantation in a Hungarian patient with cystic fibrosis]. AB - INTRODUCTION: Progressive lung disease and respiratory insufficiency is the leading cause of early death for patients with cystic fibrosis. METHOD: Successful heart-lung transplantation was performed in the 24 years old man with cystic fibrosis for end stage lung diseases and for concomitant dilatative cardiomyopathy. RESULTS: Six months following the operation the patient returned to full-time his job, and leads an active life like other fellow-men of his age. CONCLUSIONS: The attention is called with this case report to a rarely necessary intervention available for Hungarian patients as well. The criteria of the lung transplantation in cystic fibrosis, the preoperative screening and the tasks of the postoperative care are summarised. PMID- 12577405 TI - [Quality of life in the course of enzyme replacement therapy for chronic pancreatitis]. AB - AIM: The ultimate goal of any treatment in chronic pancreatitis is to maximize the patient's quality of life. The authors evaluated the QoL of patients with chronic pancreatitis prior to and after pancreatic enzyme replacement therapy in a prospective, multicenter, follow-up study. PATIENTS AND METHODS: Two groups of patients were evaluated. Group 1: 31 patients with newly diagnosed chronic pancreatitis who had never been treated with pancreatic enzyme preparations; Group 2: 39 patients whose disease was diagnosed on average 3.4 years before the start of the study. The latter group of patients had undergone pancreatic enzyme replacement therapy, but during the follow-up this proved to be insufficient. The dose of pancreatic enzyme replacement therapy was tailored in accordance with the degree of pancreatic exocrine insufficiency measured by means of exocrine pancreatic function tests. RESULTS: A significant gain in body weight and a significantly reduced defecation rate were found in both groups 1 month after the beginning of the pancreatic enzyme replacement therapy as compared to the pretreatment values. The prevalence of general and disease-specific symptoms, and the intensity of pain were reduced in both groups after 1 month of enzyme substitution therapy. The working ability, the emotional functioning, the financial strain and the overall QoL score were improved significantly in both groups, while the cognitive functioning was found to be significantly improved during the follow-up only in Group 1. The overall increase in the QoL score correlated significantly with the increase in body weight and the decrease in defecation number in both groups. CONCLUSIONS: Individually-tailored enzyme replacement therapy improved the QoL, reduced the extents of steatorrhea and pain, increased the body weight, not only in the untreated chronic pancreatitis patients, but even in the inadequately treated group. PMID- 12577406 TI - [Gynecologic and obstetric complications in eating disorders]. AB - OBJECTIVE: This report summarizes recent clinically relevant findings concerning gynecological psychosomatic symptoms of eating disorders the implications for psychiatric diagnosis and management. METHOD: The author conducted selective reviews of the literature from the last 15 years. RESULTS: Malnutrition, low body weight are also an important cause of anovulation and amenorrhoea. Eating disorders such as anorexia nervosa, bulimia nervosa are associated with potential negative consequences during pregnancy, including higher rates of miscarriage, low birth weight, obstetric complications and post partum depression. CONCLUSION: The most suitable treatment for infertility secondary to weight related amenorrhoea or anovulation is therefore dietary rather than induction of anovulation. This highlights the importance of early detection and treatment by primary care teams. There is a need to develop preventive intervention strategies. PMID- 12577407 TI - [Autogenous bone versus beta-tricalcium phosphate graft alone for bilateral sinus elevations (2-3D CT, histologic and histomorphometric evaluations)]. AB - INTRODUCTION: When the maxilla is edentulous and the alveolar process is extensively absorbed, a dental root can be implanted only after the implantation of bone or a bone-substitute. Only in this way can the subjective and objective negative features associated with a removable prosthesis be avoided. Many forms of bone-substitutes are known. Freely taken bone from the patient generally serves as the gold standard for the classification of bone-substitutes. AIMS: The aim of our work was to compare two materials (the patient's own bone and beta tricalcium phosphate) in the same patient. METHODS: Ten patients were selected who for some reason did not want or could not wear a removable prosthesis. The maxilla was so atrophied that bone or bone-substitute implantation was necessary before the dental root could be implanted. The maxilla had to be elevated from inside (sinus elevation) and thickened from outside (onlay-plasty). Bone was taken in the usual manner from the hipbone. For the internal elevation, such autogenous bone was utilized on one side, and beta-tricalcium phosphate granulate on the other. The formation of new bone and the rate of bone formation were followed by clinical methods and by radiological, histological, and histomorphometric examinations. RESULTS: The implantation succeeded clinically in all ten patients: one year later they all received a fixed bridge. The radiological and histological examinations demonstrated good bone formation on both sides. As concerns the rate of formation of new bone, there was practically no difference after the implantation of autogenous bone or beta-tricalcium phosphate. CONCLUSIONS: This study has therefore provided further evidence that, when certain bone deficiencies are to be eliminated, the unpleasant phenomena accompanying the removal of the patient's own bone can be avoided through the use of new synthetic materials. Accordingly, when comparing the present results with the findings of other authors, beta-tricalcium phosphate may be considered a good graft material even without autogenous bone. PMID- 12577408 TI - [Appointment of private lecturers during the first hundred years of the medical faculty of Pest (1770-1870)]. PMID- 12577409 TI - [First rate pediatrician from the second rank: Ferenc Torday (1871-1942)]. PMID- 12577410 TI - [Radioiodine treatment of differentiated thyroid carcinomas: residual ablation]. PMID- 12577411 TI - [Modernization of Chinese materia medica]. PMID- 12577412 TI - [Primary discussion on treatment of schizophrenia by integrated Traditional Chinese and Western medicine]. PMID- 12577413 TI - [Clinical and experimental study on effect of shuanghuang shengbai granule on myelosuppression induced by chemotherapy and ultrastructure observation of bone marrow in mice]. AB - OBJECTIVE: To observe the leukocyte increasing effects of Shaunghuang Shengbai Granule (SHSBG) in tumor patients treated by chemotherapy (CT) and its function on bone marrow hematopoietic microenvironment in mice. METHODS: Patients of non small-cell lung cancer and breast, gastric or intestinal cancer, who were retreated with CT, were enrolled and divided into 4 groups randomly. The 28 cases in the treated group 1 received SHSBG and CT simultaneously. The 27 cases in the treated group 2 also received firstly CT, and SHSBG started when WBC count lowered to less than 4 x 10(9)/L after CT. The control group 1 and 2 (n = 33 and n = 24) was treated by the method similar to that applied to the treated group 1 and 2 respectively but with Rubidate instead of SHSBG. Experimental study of observing bone marrow ultrastructure in mice was also conducted with electron microscopy. RESULTS: The total leukocyte increasing effective rate occurred in the treated group 1 was 75.00%, that in the treated group 2 was 88.89%, in the control group 1 was 54.55% and in the control group 2 was 58.33%. There was significant difference between the treated groups and the control groups (P < 0.01). Experimental study showed that SHSBG has good bone marrow hematopoietic microenvironmental protecting and improving effect in mice. CONCLUSION: SHSBG has obvious protecting and treating effect on CT caused bone marrow suppression in tumor patients. PMID- 12577414 TI - [Clinical observation on nasopharyngeal carcinoma treated with combined therapy of radiotherapy and ginseng polysaccharide injection]. AB - OBJECTIVE: To observe the effect of radiotherapy (RT) combined with Ginseng polysaccharide (GSP) injection in treating nasopharyngeal carcinoma (NPC) and its influence on immune function. METHODS: One hundred and thirty-one NPC patients were randomly divided into two groups, 64 cases in the RT-GSP group treated with RT-GSP and 67 in the control group treated with conventional therapy, to observe the local cancer remission rate, 1-year total survival rate, no tumor survival rate and no remote metastasis survival rate. Moreover, the changes of T lymphocyte subsets, natural killer (NK) cell activity and lymphocyte activated killer (LAK) cell activity before and after treatment were observed. RESULTS: Clinical examination conducted 3 months after treatment showed that the complete remission rete in the RT-GSP group was 96.6%, and in the control group 93.3%, the complete remission rate of cervical node metastatic tumor in the two groups was 85.7% and 78.0% respectively, and the NPC CT remission rate 60.3% and 51.7% respectively. Re-examination carried out 1 year after RT treatment showed that the total survival rate in the two groups was 100% and 96.5%, no tumor survival rate 84.4% and 74.6%, and no remote metastasis survival rate was 93.8% and 88.1% respectively. The activity of NK cell and LAK cell as well as T3, T4 value in peripheral blood increased significantly in the RT-GSP group (all P < 0.05) after treatment, while in the control group, activity of NK cell and LAK cell changed insignificantly after treatment, and T3, T4 value lowered significantly (P < 0.05). No toxic-adverse reaction of GSP was found. CONCLUSION: GSP has certain improving immune function effect in NPC patients during RT, it could also eliminate the occurred adverse reaction to RT and the general condition of patients. PMID- 12577415 TI - [Clinical study on post-operational anti-metastasis effect of xiaoliu pingyi mixture]. AB - OBJECTIVE: To observe the effect of combined use of chemotherapy (CT) and Xiaoliu Pingyi Mixture (XLPYM) in anti-metastasis, anti-relapse and tumor inhibition in patients after surgical operation. METHODS: Nienty-six patients of common tumor after operation were divided into 2 groups, the 58 patients in the observed group treated with CT plus XLPYM and the 38 patients in the control group treated with CT alone. After 2 courses of treatment, the status of 1 year remote metastasis and in local relapse of tumor were observed. For patients of relapse or metastasis, another 2 courses of treatment was given, then the changes of tumor size, quality of life, body weight, peripheral blood cells, immunity and hemorrheology were estimated and compared between the two groups. RESULTS: The remote metastatic rate and local relapse rate in the observed group were lower than those in the control group significantly (8.62% vs 44.74% and 15.52% vs 60.53%, P < 0.01). For patients of metastasis or relapse, the effective rate in the observed group was 46.24% and that in the control group was 24.00%, showing significant difference between the two groups (P < 0.05). Clinical observation also displayed that XLPYM could elevate the quality of life, regulate the disordered T-lymphocyte subsets, improve the hemorrheologic character and reduce the toxic-adverse reaction of CT in patients. CONCLUSION: XLPYM in combining with CT has effects of tumor inhibition, anti-metastasis and anti-relapse in treating patients of common tumor in post-operational period. PMID- 12577416 TI - [Effect of langchuangding on serum soluble interleukin-2 receptor and neopterin level in patients of systemic lupus erythematosus]. AB - OBJECTIVE: In order to explore the therapeutic mechanism of Langchuangding (LCD) in treating systemic lupus erythematosus (SLE) by observation of its effect on serum soluble interleukin-2 receptor (sIL-2R) and neopterin level in SLE patients. METHODS: Forty-five patients of SLE were randomly divided into two groups, the 25 cases in the treated group were treated with LCD plus glucocorticoid (GCC) and the 20 cases in the control group were treated with GCC alone. The levels of sIL-2R and neopterin were observed before and after treatment by double antibody sandwich ELISA. RESULTS: SLE patient showed higher levels of sIL-2R and neopterin than normal, and being higher in active stage than in remission stage. The level of neopterin was positively correlated with erythrocyte sedimentation rate (ESR) (r = 0.86, P < 0.01). After treatment, the levels of sIL-2R and neopterin were lowered significantly in the treated group than those in the control group (P < 0.05). CONCLUSION: LCD could effectively reduce the serum levels of sIL-2R and neopterin in SLE patients, which would be beneficial to cellular immune functional regulation and induce the disturbed internal environment of immunity to homeostasis. PMID- 12577417 TI - [Effect of sanwu hypotensive decoction on blood pressure and lymphokine activated killer cell in patient of primary hypertension and spontaneously hypotensive rats]. AB - OBJECTIVE: To investigate the effect of Sanwu hypotensive decoction (SWHD) on blood pressure (BP) and lymphokine activated killer cell (LAK) and possible mechanism. METHODS: Thirty mild hypertension patients were treated with SWHD for 8 weeks, the levels of BP, proliferative ability and activity of LAK cell, SOD like substance expressed by LAK cell and its radical scavenger ability before and after treatment were observed using randomized, single blinded, self-control, paralled assay, and compared with other 30 patients treated with captopril for control. In an experimental study, the vasodilatory response of thoracic aortic ring to acetylcholine and nitroglycerine, and effect of LAK cell, standard reagent of SOD, SWHD on the response were observed and compared in spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKR). RESULTS: After SWHD treatment, along with lowering of BP, the proliferative ability, activity SOD like substance expressed by LAK cell and its radical scavenger ability increased significantly. CONCLUSION: SWHD is not only an effective hypotensive agent, but also an immunoregulator. PMID- 12577418 TI - [Treatment of lumbar intervertebral disk prolapse by estradural injection of mailuoning and its influence on dynorphin A in cerebrospinal fluid]. AB - OBJECTIVE: To explore the modulating effects of extradural injection of Mailuoning on analgesic function. METHODS: Twenty patients of lumbar intervertebral disk prolapse were treated by extradural injection of Mailuoning, the spontaneous release of dynorphin A (Dyn-A) in cerebrospinal fluid (CSF) before and after treatment were measured and compared with that of the 20 cases in the control group to determine the therapeutic effect of Mailuoning. RESULTS: After treatment, the concentration of Dyn-A in CSF in the treated group increased remarkably, but displayed no significant difference to normal healthy group (P > 0.05), while that in the control group there was significant difference between them (P < 0.01). CONCLUSION: Mailuoning has the modulatory effect on analgesia function and lumbar intervertral disk prolapse was treated by means of its Dyn-A elevating action. PMID- 12577419 TI - [Observation on effect of supplementary treatment by Astragalus injection in treating senile pulmonary tuberculosis patients]. AB - OBJECTIVE: To observe the clinical therapeutic supplementary effect of Astragalus injection (ASI) as anti-tuberculosis agents in treating senile tuberculosis (ST). METHODS: Seventy-six ST patients were divided according to their hospitalization order into two groups randomly, 39 in the ASI group and 37 in the control group. The anti-tuberculosis regimen applied on all patients were HRE (S)Z for first treated patients and KHZ1321 TH for retreated patients. In the ASI group, ASI was given additionally by adding ASI 20 ml into 500 ml 5% glucose solution for intravenous dripping, once a day. The therapeutic course was 2 months. The changes of focal size, bacteria in sputum, and erythrocyte immune function (EIF) were observed before and after treatment, and the EIF obtained from 30 healthy subjects was taken for control. RESULTS: EIF in patients of both groups was lower than that in healthy subjects (P < 0.01). Rosette rate of RBC-C3b receptor in both groups was all increased after treatment, the increment was higher in the ASI group than that in the control group significantly (P < 0.01). After 2 months ASI treatment, the effective rate of focal absorption examined by X-ray was 84.6% and the negative conversion rate of bacteria in sputum was 79.4%. CONCLUSION: ASI has the effect of elevating erythrocyte immunity in senile pulmonary tuberculosis patients, it is able to enhance the therapeutic effect of treatment. PMID- 12577420 TI - [Influence of Salvia miltiorrhizae and Astragalus membranaceus on hemodynamics and liver fibrosis indexes in liver cirrhotic patients with portal hypertension]. AB - OBJECTIVE: To observe the influence of Salvia miltiorrhiza (SM) and Astragalus membranaceus (AM) on hemodynamics and liver fibrosis indexes in patients of liver cirrhosis with portal hypertension. METHODS: Eighty-four cases of liver cirrhosis were enrolled and divided randomly into two groups, 42 in each. The control group was treated with conventional therapy and the tested group treated with SM and AM. The parameters, including diameter of portal vein and splenic vein (Dpv and Dsv), speed of blood flow in portal vein and splenic vein (Spv and Ssv), quantity of blood flow in portal vein and splenic vein (Qpv and Qsv) as well as liver fibrosis indexes, such as HA, PC III and LN, were determined before, 1, 2 and 3 months after treatment. RESULTS: After treatment, in the tested group, Dpv and Dsv decreased, Spv and Ssv increased, and Qpv and Qsv reduced, showing a significant difference in comparison with those in the control group (P < 0.05 or P < 0.01). The liver fibrosis indexes were improved significantly in the tested group, also showed significant difference from those in the control group (P < 0.01). CONCLUSION: SM and AM could improve portal hypertension effectively in liver cirrhosis patients, one of the mechanism may be related with the improvement of liver fibrosis. PMID- 12577421 TI - [Regulative effects of bailong and Hexamethylene bisacetamide on growth and phenotype of human gastric cancer cells in cell cycle]. AB - OBJECTIVE: To investigate and compare the common effect of Bailong, a Chinese herbal preparation, and Hexamethylene bisacetamide (HMBA), a western cell differentiation inducer, on growth regulation and phenotype of human gastric cancer (MGC80-3) cells in different phases of cell cycle. METHODS: Synchronized MGC80-3 cells in monolayer cultures with nitrous oxide under high pressure were double blocked with overdosage of TdR in the 4 cellular phases (G1, S, G2 and M), and then collected and treated with Bailong and 5 mmol/L HMBA respectively. RESULTS: Both medicines could suppress the cell proliferation, soft agar cloning growth and microfilament assembling of different phases of cells, the effect on cells of G1 phase was the most significant one. CONCLUSION: Bailong and HMBA had significant common characteristics in inducing cell differentiation of MGC80-3 cells in different phases of cell cycle. PMID- 12577422 TI - [Imitative restenosis after percutaneous transluminal coronary angioplasty prevented by buyang huanwu decoction in rabbits]. AB - OBJECTIVE: To examine the preventive effect of Buyang Huanwu Decoction (BYHWD) on imitative restenosis after percutaneous transluminal coronary angioplasty (PTCA) in diet-induced atherosclerotic rabbits. METHODS: Diet-induced atherosclerotic rabbits were randomly allocated into group 1 (n = 24, fed with BYHWD one week before and 4 weeks after the operation, 2 ml/kg per day, containing herbal drug 8.4 g/ml) and group 2 (n = 20, as control). Balloon angioplasty was performed in the abdominal arota to mimic coronary angioplasty. RESULTS: Sixty days after operation: (1) Incidence of hyperplasia of the intima was 7/24 (29.2%) in group 1 vs 17/20 (85.0%) in group 2, P < 0.01; (2) Comparison of area of hyperplastic intima was (0.54 +/- 0.21) mm2 vs (1.67 +/- 0.75) mm2, P < 0.01; (3) Area of hyperplastic intima/area of lumen (limited by the internal elastic membrane was (18.0 +/- 7.2)% vs (56.0 +/- 17.1)%, P < 0.01; (4) Intact lumen/dilated lumen was (0.74 +/- 0.03) vs (0.35 +/- 0.07), P < 0.01; (5) Effect of serum taken from group 1 on cultured smooth muscle cells showed by 3H-TdR method was (4110.14 +/- 1977.01) cpm, which was significantly less than that of serum taken from group 2 [(7960.14 +/- 2802.59) cpm, P < 0.05]. CONCLUSION: BYHWD has significant preventive effect on the imitative post PTCA restenosis in diet-induced atherosclerotic rabbits. PMID- 12577423 TI - [Study on modulation effect of qi regulating and blood activating drugs on mechanism of immunity and hemorrheology in stressed rats]. AB - OBJECTIVE: To explore the modulation effect of Chinese herbal medicines for Qi regulating and blood activating (QRBA) on mechanism of immunity and hemorrheology in stressed rats. METHODS: The influence of QRBA on plasma noradrenaline (NA), proliferation of splenic cells and hemorrheologic properties were observed in stressed rats. RESULTS: The Chinese herbal medicines for soothing Liver to regulate Qi, activating blood circulation to remove stasis and QRBA drugs could antagonize in various degrees the changes caused by stress. Among them QRBA drugs was the best, it could reduce the level of plasma NA (P < 0.01), modulate the blood hyperviscosity induced by exogenous NA (P < 0.05) and enhance proliferation of splenic cells (P < 0.01) in rats. CONCLUSION: QRBA drugs strengthen immune function and restore hemorrheologic properties by reducing NA content in organism. PMID- 12577424 TI - [Qi deficiency and blood stasis and circulatory renin-angiotensin system as well as plasminogen activator inhibitor activity in rats with cardiac qi deficiency syndrome]. AB - OBJECTIVE: To explore the relationship between cardiac Qi deficiency Syndrome and effect of circulatory renin-angiotensin system (RAS) and endogenous fibrinolytic system in rats with heart failure (HF). METHODS: Plasma RAS levels and plasminogen activator inhibitor (PAI-1) activity were measured in 2 groups of rats: the sham-operated group (Group A) and the HF with Heart Qi deficiency Syndrome model group (Group B) established by making arteriovenous fistula. Using radioimmunoassay and chromatography, plasma renin activity (PRA) and angiotensin II (Ang II) levels and PAI-I activity were determined before, immediately after and 30 days after the surgical operation respectively. RESULTS: After the operation was completed in Group B the left ventricular dysfunction revealed, left ventricular systolic pressure decreased and end diastolic pressure increased (P < 0.05), PRA and Ang II level and PAI-1 activity increased significantly, as compared with those in Group A and before modeling (P < 0.05). With cardiac function improved, the plasma PRA, Ang II level and PAI-1 activity lowered in Group B comparing with those immediately after modeling (P < 0.05). CONCLUSION: The activation of RAS is associated with the endogenous fibrinolyticim balance, and play an important role in endogenous fibrinolytic system dysfunction in HF with Heart Qi deficiency Syndrome and Qi deficiency-blood stasis, which was helpful to explain that cause of hypercoagulant state in HF patients and increase the risk of suffering from embolism-thrombotic diseases, and might be the pathogenetic basis of Heart Qi deficiency induced Qi deficiency-blood stasis. PMID- 12577425 TI - [Immunomodulatory activity and anti-tumor activity of Oldenlandia diffusa in vitro]. AB - OBJECTIVE: To study the effect of Oldenlandia diffusa (OD) on lymphocytes and tumor cells in vitro. METHODS: Effects of OD extract (ODE) on proliferation of spleen cells of mice and the phagocytosis of monocytes to tumor cells using 3H incorporation were analysed, and effect on specific activity of natural killer (NK) cells to human and mice's tumor cells was determined using free 51 Cr experiments, effects on B cells' antibody production and cytokine of monocyte production were investigated by ELISA and biological method, the chemical composition of OD was analyzed by chromatography, protease digestion and sugar decomposition (NaIO4) test. RESULTS: ODE could vigorously promote the proliferative activity of spleen cells in mice, specific lethal activity of human and mice's NK cell to tumor cells, B cells' antibody production, monocytes' cytokine production and its phagocytosis to remove the tumor cells. By means of chromatography, protease E digestion and NaIO4 treatment, the result of chemical composition analysis showed that the component of OD was 90 kDa glycoprotein. CONCLUSION: ODE has immuno-modulating activity and anti-tumor activity in vitro through stimulating the immune system to kill or engulf tumor cells, which could be used clinically for immune function modulation and to treat tumor and other diseases. PMID- 12577426 TI - [Experimental study on protective effective of congsheng capsule in acute cerebral ischemia and improvement of cerebral blood flow and energy metabolism]. AB - OBJECTIVE: To investigate the protective effect of Congsheng capsule (CSC) on acute cerebral ischemia in mice and rats. METHODS: Cerebral ischemia model was established by adding FeCl3 to block the blood flow of unilateral middle cerebral artery (MCA) for 30 mins. Acute incomplete cerebral ischemia was induced by ligation of bilateral carotid arteries in rats. Bilateral carotid arteries of old mice were repeatedly ischemia and reperfusion combined with caudal bloodletting to build the cerebral ischemia model. The effect of CSC on neurologic symptom, cerebral infarction size, cerebral water content, cerebral blood flow and cerebral tissue energy metabolism were observed. RESULTS: CSC 1, 3, 6 g/kg obviously reduce the size of cerebral infarction and cerebral water content, markedly improve the neurological symptoms and cerebral blood flow; 3, 6 g/kg significantly ameliorate the energy burden. CONCLUSION: CSC has anti-cerebral ischemia function through increasing blood supply of cerebral tissue and improve the energy metabolism. PMID- 12577428 TI - [Progress in the study of the treatment of nephropathy with Astragalus and Angelica and their therapeutic mechanism]. PMID- 12577430 TI - [Antifebrile effect of naoreqing oral liquid on post-operational fever of glioma or meningioma patients]. AB - OBJECTIVE: To observe the antifebrile effect of Naoreqing (NRQ) oral liquid on fever in patients after glioma or meningioma operation. METHODS: Patients were divided into 3 groups, the TCM group treated with NRQ, the TCM-WM group treated with NRQ plus western medicine and the WM group treated with western medicine alone. The treatment was applied for 7 successive days. The mean time of fever and fever subsidence, body temperature before and after treatment were recorded and compared. RESULTS: The antifebrile effect in the TCM and the TCM-WM group was superior to that in the WM group significantly (P < 0.01), and the difference between the TCM group and the TCM-WM group was insignificant. CONCLUSION: NRQ can evidently reduce the body temperature of patients after brain tumor operation, its antifebrile effect is irrelevant to the kind of tumor being glioma or meningioma. PMID- 12577431 TI - [Syndrome differentiation patterns for peri-operative coronary heart disease patients of coronary artery bypass graft]. AB - OBJECTIVE: To explore the patterns of Syndrome Differentiation of coronary artery bypass graft (CABG) on the peri-operative coronary heart disease (CHD) patients. METHODS: One week after operation, thirty-seven CHD patients, who received CABG of internal mammary artery or great saphenous vein under conventional general anesthesia with low or middle temperature extracorporeal circulation were differentiated as various syndromes, the pre- and post-operational ECG, color Doppler echocardiography as well as during and after operation. The hemodynamic parameters were monitored. RESULTS: In the CHD patients, 64.9% were differentiated as Qi-Yin deficiency, 67.6% were complicated with Phlegm Syndrome and 62.2% with Stasis in blood, suggesting that Qi-deficiency, phlegm and stasis are the basic pathogenesis in patients after CABG. Moreover, the peri-operative Syndrome was correlated with the condition of coronary arterial lesions, heart and lung functions before operation, and the time for extracorporeal circulation during the operation. CONCLUSION: TCM Syndrome Differentiation conducting in peri operative stage might be useful to explore the patterns of Syndrome alteration which provided a basis for preventing peri-operational complication and elevating success rate of operation. PMID- 12577432 TI - [Comparative clinical study on treatment of varicocele caused infertility by tongling granule and surgical operation]. AB - OBJECTIVE: To observe the effect of Tongjing granule (TJG) in treating varicocele caused infertility. METHODS: Comparative observation was carried on 75 cases of male infertility caused by moderate or severe varicocele with abnormal semen, they were divided into two groups: the 44 patients in the TCM group treated with TJG and the 31 patients in the operation group treated by surgical operation (high ligation of spermatic vein and/or inferior epigastric venous bypass). All patients were followed up periodically to observe the amount, survival rate and activity of sperm by computerized automatic seminal analyser (CASA), as well as the occurrence of pregnancy in their partner. RESULTS: No significant difference was found in various parameters between the two groups. After treatment, the improvement of seminal density, amount of active sperms and forward moving sperms were lower in the TCM group than that in the operation group, but according to CASA, the improvement of TCM group is better than that of operation group. There was insignificance in the operation group between pre- and post-treatment, P > 0.05. CONCLUSION: TJG could treat the varicocele caused infertility with the clinical effect similar to the surgical operation, and was superior in improving motility of sperm. It indicates that surgical operation could only improve the local lesion, but TJG could modulate the general condition of patients also. PMID- 12577433 TI - [Clinical study on treatment of oviduct obstruction by integrative traditional Chinese and Western medicine]. AB - OBJECTIVE: To find an effective and practical treatment of oviduct obstruction (OvO). METHODS: One hundred and twenty OvO patients were randomly divided into three groups, the TCM-WM group, treated with integrative traditional Chinese and western medicine, the TCM group treated with Chinese herbal medicine alone and the WM group treated with western medicine alone. The therapeutic effect as well as the effect of treatment on serum C-reactive protein (CRP) and interleukin-1 beta (IL-1 beta) were observed. RESULTS: After treatment, the fallopian tube patency rate was 86.7% and the pregnant rate 85.0% in the TCM-WM group, while in the TCM group was 66.7% and 63.3% respectively, and in the WM group 53.3% and 50.0% respectively. Comparison among the three groups showed that the effect in the TCM-WM group was significantly superior to that in the other two groups (P < 0.01). The levels of CRP and IL-1 beta were all lowered after 3 courses of treatment, and the effect was more evident in the TCM-WM group (P < 0.01). CONCLUSION: TCM-WM treatment is a good and practical method in treating oviduct obstruction. PMID- 12577434 TI - [Clinical and experimental study on treatment of infantile mycotic enteritis by jiechang mixture]. AB - OBJECTIVE: To evaluate the therapeutic effect and mechanism of Jiechang Mixture (JCM) in treating infantile mycotic enteritis. METHODS: The children patients confirmed to be suffered from mycotic enteritis were divided randomly into two groups: the 40 cases in the treated group treated with JCM and the 20 cases in the control group treated with fungicidin. The time of stool forming, diarrhea relieving and fungi vanishing were observed. The promoting effect of JCM on small intestine motility and the inhibition of JCM on Candida albicans in stool culture were studied experimentally. RESULTS: The time of stool forming, diarrhea relieving and fungi vanishing in the treated group were significantly shorter than those in the control group (P < 0.01), and JCM also showed a better effect in improving clinical symptoms and signs of patients than the control. Results of experimental study showed that JCM could abate the ink evacuation of small intestine in mice. The fungi inhibitory test suggested that Candida albicans was susceptible to JCM. CONCLUSION: JCM is an effective herbal medicine in treating infantile mycotic enteritis. PMID- 12577435 TI - [Clinical study on treatment of gastric ulcer with qingwei zhitong pill]. AB - OBJECTIVE: To observe the therapeutical effect of Qingwei Zhitong pill (QWZTP) in treating gastric ulcer and clearance of the Helicobacter Pylori (HP). METHODS: Patients in the treated group (n = 60) and the control group (n = 60) were treated with QWZTP and Sifangwei tablet separately to observe the therapeutic effect of treatment on ulcer niche, TCM Syndromes and HP. RESULTS: The effective rate on ulcer niche evaluated by gastroscope was 86.67% in the treated group and 71.67% in the control group, the comparison between the two groups showed a significant difference, P < 0.05. The effective rate on improving TCM Syndrome in the two groups was 91.67% and 88.33% respectively, the difference between two groups was insignificant (P > 0.05). The HP clearance rate in the treated group was 47%. CONCLUSION: QWZTP has good effect in treating gastric ulcer and clearance of HP. PMID- 12577436 TI - [Effect of shenqi fuzheng injection on immune function in gastric carcinoma patients in post-operational and chemotherapeutic period]. AB - OBJECTIVE: To investigate the effect of Shenqi Fuzheng injection (SQFZI) on immune function in gastric carcinoma patients in post-operational and chemotherapeutic period. METHODS: One hundred and twenty-three gastric carcinoma post-operational patients and 36 cases of gastric carcinoma patients on chemotherapy, according to their using SQFZI or not, parameters of immune function, including red blood cell C3b receptor rosette (RBC-C3bRR) and immune complex rosette (RBC-ICR), T-lymphocyte subsets and NK cell activity were determined. RESULTS: RBC-C3bRR, CD3, CD4 and NK cell activity were significantly higher, but RBC-ICR was lower in patients of post-operational period treated with SQFZI than those in patients untreated with SQFZI (P < 0.01). The same difference also revealed between the patients in chemotherapeutic period treated and untreated with SQFZI (P < 0.01). CONCLUSION: The immune function of gastric carcinoma patients in post-operational or chemotherapeutic period could be effectively improved by addition of SQFZI. PMID- 12577437 TI - [Clinical and experimental study on treatment of moderate and advanced malignant tumors with tianfoshen oral liquid]. AB - OBJECTIVE: To investigate the clinical efficacy of Tianfoshen oral liquid (TFS) in treating moderate and advanced malignant tumors and its mechanism. METHODS: Therapeutic effect of TFS in treating 71 patients of malignant tumor was analyzed with the criteria, including quality of life, 3-year survival rate and immune function. And experimental studies of inhibitory effect on tumor clone primordial cells and tumor growth rate of TFS on human gastric tumor (MGC803) cell, human liver cancer (SMMC7721) cell and mice galactophore cancer (EMT6) cell by colony forming method and dye exclusion test respectively were also conducted. RESULTS: Clinical study showed that in the 71 cases treated, the total remission rate was 45.1%, the effective rate 71.8%, with improvement in quality of life and immune function, the 1-, 2- and 3-year survival rate being 78.5%, 38.5% and 10.8% respectively and the mean survival time 24.2 months. Experimental study showed that TFS could kill the cancer cells directly, inhibit the proliferation of single clonogenic cell, and had a broad-spectrum dose-dependent inhibitory action on various tumors with significant difference in comparing with the effects of the control (P < 0.05). CONCLUSION: TFS had obvious therapeutic effect to moderate and advanced tumors, its anti-tumor effect was related to the enhancement of immune function and tumor inhibiting or direct killing action. PMID- 12577438 TI - [Clinical study on treatment of protracted pain by continuously administering different dose of fentanyl by ocupoint embedding micropump]. PMID- 12577439 TI - [Study on effect of emodin on TGF beta 1 expression in pancreatic tissue of rats suffering from acute pancreatitis]. AB - OBJECTIVE: To investigate the effect and mechanism of emodin on pancreatic repairing and remodeling in treating acute pancreatitis by analyzing the change of cytokine transforming growth factor beta 1(TGF beta 1) gene expression, DNA synthesis and protein content in pancreatic tissue. METHODS: Acute pancreatitis was induced in rats by intraperitoneal infusion of caerulein, treated or untreated by emodin. The animals were sacrificed at 6, 24, 48, 72 or 96 hrs after treatment separately. The mRNA expression of TGF beta 1, DNA synthesis and total protein content in pancreatic tissue were tested by reverse transcription polymerase chain reaction (RT-PCR), 3H-thymidin method and Lowry's method respectively. RESULTS: Serum amylase level was decreased significantly after emodin treatment. TGF beta 1 mRNA expression was undetectable in the intact pancreas or in 6 hrs after pancreatitis induction in the non-treated group, but revealed at 24 hrs after and reached the peak at 72 hrs later. However, in the emodin treated group, TGF beta 1 mRNA expression was detected at 6 hrs after treatment, with a higher level in 24 hrs and 48 hrs as compared with the non treated group, and reaching the peak at 48 hrs after treatment. Moreover, the DNA synthesis and total protein content in pancreatic tissue decreased significantly at 72 hrs and 48 hrs after induction respectively, but both parameters increased significantly in the emodin treated group 96 hrs after treatment. CONCLUSION: The mechanism of emodin in treating acute pancreatitis might be by way of enhancing cytokine TGF beta 1 gene expression, regulating cell growth and differentiation, stimulating the formation of extracellular matrix components, increasing DNA synthesis and protein content, and to take part in pancreatic repairing and remodeling. PMID- 12577440 TI - [Effect of wuzi yanzong pill and its disassembled prescription on mitochondrial DNA deletion, respiratory chain complexes and ATP synthesis in aged rats]. AB - OBJECTIVE: To investigate the effect of Wuzi Yanzong Pill (WZYZP) and its disassembled prescription on mitochondrial DNA deletions, respiratory chain complexes and ATP synthesis in aged rats. METHODS: Animal experiments with polymerase chain reaction (PCR), enzyme kinetics and bioluminescence technique were conducted. RESULTS: WZYZP and its disassembled prescription of Fructus Lycii and Semen Cuscutae could reduce the mitochondrial DNA deletions and raise the activities of mitochondrial respiratory chain complexes I, IV and adenosine triphosphate (ATP) synthesis in aged rats' brain; Fructus Lycii and Semen Cuscutae could also reduce the mitochondrial DNA deletions in aged rats' heart (P < 0.05, P < 0.01). CONCLUSION: WZYZP, Fructus Lycii and Semen Cuscutae have protective effect on oxidative damage of mitochondrial DNA in aged rats. PMID- 12577441 TI - [Protective effect of yixinkang capsule containing serum on oxygen stress damaged mitochondrial membrane potential in guinea-pig myocytes]. AB - OBJECTIVE: To explore the protective effect and mechanism of Yixinkang (H303) capsule, a composite preparation of Chinese herbal medicine for antagonizing myocardial ischemia, containing serum on oxygen stress damaged mitochondrial membrane potential (MMP) in guinea-pig myocytes. METHODS: Myocyte damage was induced by incubating myocytes with hydroxy free radical (Fe2+/H2O2) generating system for 30-60 mins, then H303-containing serum was added in to observe the protective effect. MMP was detected by laser scanning confocal microscopy. RESULTS: After oxygen stress, the cytochrome oxidase (CCO) activity and MMP in mitochondria significantly decreased. These changes could be significantly lessened by H303-containing serum, demonstrated as obviously increase of CCO activity and myocyte survival rate. H303-containing serum also showed a significant protective effect on membrane potential of myocytes. CONCLUSION: Oxygen stress could cause decrease of MMP in myocytes and H303-containing serum could protect it to maintain the membrane stability of mitochondria. PMID- 12577442 TI - [Effects of quanjia yangshen capsule on rate of polychromatic erythrocytic micronucleus formation and peripheral blood picture in mice treated by cyclophosphamide]. AB - OBJECTIVE: To observe the effect of Quanjia Yangshen capsule (QJYS) on cyclophosphamide (Cy) induced teratogenesis and bone marrow in mice and to explore the mechanism of its clinical therapeutic effect. METHODS: Mice were separately administered with QJYS solely, Cy solely and QJYS + Cy and the effect on polychromatic erythrocytic (PE) micronucleus formation rate in the bone marrow of mice, and peripheral blood picture were observed and compared with the control group. RESULTS: High dosage of QJYS has no apparent effect on PE micronucleus formation rate and peripheral blood picture in the normal mice; Cy can obviously raise PE micronucleus formation rate (P < 0.01), and lowered WBC, RBC and Hb obviously (P < 0.01) in mice. After taking QJYS, the Cy induced micronucleus formation rate increasing and blood picture parameters lowering were controlled. The improvement in the high and middle dose QJYS group was significantly higher in comparing to that in the sole Cy group (P < 0.01); while the low dose QJYS group in comparing with Cy group also showed significant improvement (P < 0.01), except Hb value. CONCLUSION: QJYS could significantly antagonize teratogenic effect and inhibition to bone marrow of Cy, i.e., play an antagonistic role against the toxic and side-effects of Cy. PMID- 12577443 TI - [Effects of kidney reinforcing and blood circulation activating Chinese medicine on morphological structure of mammary glands in models of mammary atrophy and hyperplasia in female rats]. AB - OBJECTIVE: To study the effects and mechanism of Kidney reinforcing and blood circulation activating Chinese medicine (K&B) on morphologic structure of mammary glands in female rat models of mammary atrophy and mammary hyperplasia. METHODS: The models of mammary gland atrophy and hyperplasia were established respectively by ovariectomy and diethylstilbestrol injection to observe the changes of gland's weight index, structure, estradiol receptor (ER) and progesterone receptor (PR) after treated by K&B. RESULTS: (1) K&B could ameliorate the atrophied structure of mammary gland with significant increase of weight index (P < 0.01), raise the number of ER and PR in mammary tissue and nuclei (P < 0.01, P < 0.05), normalize the ratios of ER and PR in plasma and those in nucleus (EnR/EcR and PnR/PcR). (2) K&B could also improve or restore the proliferated structure of mammary gland and its weight index as well as the EnR/EcR and PnR/PcR ratios. CONCLUSION: K&B has the effects in preventing and improving the mammary gland atrophy induced by hyposecretion of sexual hormone and promoting the development of mammary gland in adult rats. Although both K&B and estrogen can improve the development of mammary gland, their mechanisms are different. There is a bright future of K&B in treating underdevelopment of mammary gland. PMID- 12577444 TI - [Seasonal variations effecting sexual functions in golden hamsters]. PMID- 12577445 TI - [Clinical and experimental studies on blood deficiency syndrome]. PMID- 12577446 TI - [Progression on treatment of nasopharyngeal carcinoma by combined therapy of radiation and Chinese herbal medicine]. PMID- 12577447 TI - [Predictors or statistics versus individual effects of chemotherapy]. PMID- 12577448 TI - [Liver biopsy in chronic hepatitis virus C infection--unnecessary, useful or useless?]. PMID- 12577449 TI - [Editorial referring to (in general) to the article: Hauer D. et al: Thyroid disease in pregnant women and its development after childbirth]. PMID- 12577450 TI - [Ethylene glycol poisoning]. PMID- 12577451 TI - [Gastrointestinal hemorrhage and endoscopic findings in patients with chronic kidney failure]. AB - Case-control study comparing 81 patients with chronic renal failure (CRF) with 58 patients without renal disease who were evaluated by endoscopy of upper and/or lower gastrointestinal tract (GIT) for gastrointestinal bleeding or dyspepsia. The success of endoscopic examinations was the same in both groups. Patients with CRF more often underwent endoscopic examination because of gastrointestinal bleeding and they presented more often by hematochezia. Patients with CRF were more often found to have hemorrhagic gastropathy or duodenopathy and angiodysplasia of upper and lower GIT. However, duodenal ulcer was significantly more common in the control group without renal disease. Incidence of Helicobacter Pylori infection was low in both groups. PMID- 12577452 TI - [Effect of transjugular portosystemic shunt on insulin resistance]. AB - Patients with cirrhosis of the liver suffer from hyperinsulinaemia and a certain degree of insulin resistance. More frequently than in the rest of the population they have diabetes. Transjugular intrahepatic portosystemic shunts (TIPS) as a therapeutic method in complications of portal hypertension lead to rapid haemodynamic changes in the liver. The objective of the submitted work was to assess whether TIPS has an impact on insulinaemia and whether it influences insulin resistance in patients with cirrhosis of the liver. GROUP AND METHODS: The authors evaluated a group of 22 patients with cirrhosis of the liver (10 diabetics and 12 subjects without diabetes) indicated for TIPS. They investigated the insulin and C-peptide concentration in blood obtained by catheterization from the hepatic and portal vein before and after TIPS and in the peripheral blood before TIPS, 1 hour, 1 day, 1 week and 1 month after TIPS. The insulin resistance was examined by the method of the hyperinsulin euglycaemic clamp (HEC) before TIPS, 1 day, 1 week, and 1 month after TIPS. The levels of C-peptide and insulin were assessed by the IRMA method. The blood sugar level in HEC was measured by means of a Hemocue apparatus. The results were evaluated by the non-parametric Wilcoxon test for two dependent samples. RESULTS: Both groups (diabetics and non diabetics) were comparable as to age, sex, etiology of liver cirrhosis and indication for TIPS. After introduction of TIPS a change of insulin clearance occurred (p = 0.01) and a change of the insulin level in the hepatic vein immediately after TIPS (p = 0.02). Insulin clearance before TIPS was 37-90% (median 54%) and after TIPS it declined to 0-79% (median 38%) (p = 0.01). Already 1 hour after the operation the authors observed a rise of the insulin level in peripheral blood as compared with baseline values (p = 0.002). Statistically significant hyperinsulinaemia persisted one month after TIPS (p = 0.005). Values of C-peptide did not change significantly in time, neither in the hepatic vein nor in the peripheral blood. On examination of IR no statistically significant changes occurred after TIPS. On evaluation of different groups of diabetics and non-diabetics the IR was more marked in patients with DM (mean M = 1.7 mg/kg/min.) than in patients without DM (3.7 mg/kg/min.) (p = 0.03). The authors did not record significant changes of IR in time in different groups. Compensation of DM was not influenced by TIPS. The fasting blood sugar levels before TIPS and 1 month after TIPS were comparable. CONCLUSION: After TIPS a rise of the insulin level in peripheral blood occurred due to the reduced insulin clearance in the liver. Despite hyperinsulinaemia which persisted for one month after the operation, the insulin resistance did not deteriorate. Compensation of diabetes was not affected by TIPS. PMID- 12577453 TI - [Importance of liver biopsy in chronic hepatitis C virus infection]. AB - INTRODUCTION: The importance of liver biopsy and knowledge of the histological activity of liver les on in chronic hepatitis C virus (HCV) infections is widely discussed recently. There are attempts to find an alternative evaluation which will make it possible to avoid liver biopsy. The crucial question in patients with chronic HCV infection is to differentiate patients with already developed liver cirrhosis from those with chronic hepatitis. OBJECTIVES: 1. To evaluate the impact of the calculation of the discrimination score of liver cirrhosis (DSC) for prediction of liver cirrhosis in the histological assessment. 2. To assess the correlation of prediction of cirrhosis liver based on clinical signs and actual histological verification. 3. To evaluate the frequency of unexpected histological findings not correlating with the clinical picture. GROUP OF PATIENTS: The group was formed by 139 patients. In all patients during the baseline examination the patient's history data were analyzed as well as possible physical signs of liver cirrhosis. In all patients also, based on laboratory values before liver biopsy, the DSC according to Bonacini was calculated. Furthermore agreement between the histological finding of liver cirrhosis and chronic hepatitis with DSC values was assessed. RESULTS: 1. Based on calculation of DSC it is possible to predict accurately the existence of cirrhosis of the liver or chronic hepatitis only in 31% patients. In 69% patients even comprehensive evaluation of the type of DSC is not a sufficient guide for assessment of the hepatic lesion. 2. Even clinical signs of cirrhosis are not a quite reliable guide for its prediction. In 8% patients of our group the histological finding of liver cirrhosis was a surprise and in 3.5% patients cirrhosis of the liver was not confirmed despite the presence of clinical signs. 3. The frequency of other histological findings participating in the development of the hepatic lesion in chronic HCV infection was minimal. In the authors group as such only steatosis and toxic damage of hepatic tissue by alcohol were identified. These findings were, however, suspected already before biopsy. Steatosis can be however considered also a manifestation of HCV infection. CONCLUSION: The results of the trial support the view that liver biopsy is in the majority of cases irreplaceable for evaluation of the severity of the hepatic affection in chronic HCV infection. PMID- 12577454 TI - [Incidence of risk factors and vascular complications in patients with newly diagnosed type 2 diabetes mellitus]. AB - The objective of the investigation was early detection of risk factors of diabetes and/or atherosclerosis and the prevalence of vascular complications of diabetes in newly diagnosed patients with type 2 diabetes in the Slovak Republic. PATIENTS AND METHODS: In the Slovak detection programme of vascular complications participated 40 diabetological out-patient departments. In the course of one year (from the beginning of June 1999 to the end of May 2000) in a total of 3424 newly diagnosed patients with type 2 diabetes, aged under 70 years the case-history was recorded, focused on risk factors of diabetes and atherosclerosis, on the patients' lifestyle, on vascular complications of diabetes and their treatment. Moreover a detailed clinical and laboratory examination was made. The authors made in addition to assessment of the blood sugar level also biochemical examinations focused on indicators of the lipid metabolism and microalbuminuria. All patients had an ophthalmological examination, and the electrocardiogram was recorded. RESULTS: As to risk factors of diabetes as many as 3259 patients (95.2%) reported a low physical activity (exercise lasting less than the minimum of 3 x 30 minutes per week) and 3001 (87.6%) had an elevated body weight, whereby 43.9% patients had a BMI of 25 to 30 kg/m2 and 43.8% were obese (BMI > 30 kg/m2). Diabetes mellitus in a first grade relative was reported by 1521 patients (44.7%). As to risk factors of atherosclerosis, the most frequent finding was hyprcholesterolaemia (values > 5.2 mmol/l) in 2262 patients (67.3%), and hypertriglyceridaemia (values > 1.7 mmol/l) in 2194 patients (66.5%). Arterial hypertension (values of systolic and/or diastolic pressure > 140/90 Hg) were recorded in 2326 (67.9%) patients. The most frequent microvascular complication of diabetes was microalbuminuria was in 20.5% patients. Ischaemic heart disease was recorded in 22.80 diabetics. Risk factors of atherosclerosis and vascular complications in newly diagnosed type 2 diabetics are closely related to the patients' body weight. CONCLUSION: The authors provided evidence of a high incidence of risk factors of diabetes and atheroscleosis in newly diagnosed patients with type 2 diabetes mellitus, and in practically every fifth patient the presence of ischaemic heart disease. Obesity is a significant adverse metabolic and vascular factor. PMID- 12577455 TI - [Parenteral glutamine does not improve the nutritional status in patients during high-dose chemotherapy and autologous peripheral stem cell transplantation]. AB - BACKGROUND AND AIM: High-dose chemotherapy is aggressive treatment modality adversely affecting both energy/protein demands and oral intake/resorption of nutrients. Aminoacid glutamine is known for its' proteoanabolic effect and as an energy source for enterocytes and immune system. Nutritional parameters have been studied in a controlled, randomised, double-blinded trial of parenteral glutamine supplementation of autologous stem cell transplant patients. METHODS: Forty consecutive patients with haematological and solid cancer and multiple sclerosis were treated from 1999 to 2001 by high-dose chemotherapy with autologous stem cell transplantation. Patients were randomly assigned either for parenteral administration of 30 g of alanyl-glutamine dipeptide (Dipeptiven, Fresenius-Kabi) or isonitrogenous glutamine-free amino acid solution from day +1 to day +14 or to discharge from hospital. Patients were closely monitored from admission to day +100. Nutritional parameters included: oral dietary intake, body weight, body composition, energy expenditure, concentration of serum proteins and nitrogen balance. Parenteral nutrition in dose of 26.5 kcal/kg and 1 g of aminoacid/kg was given to patients who did not reach adequate oral intake for 5 days and withheld after three consecutive days of adequate intake. RESULTS: Nutritional assessment on admission differed according to the method used but no parameter of nutrition predicted the clinical course of treatment. Inadequate oral intake period lasted (mean +/- SD) 6.8 +/- 5.9 days, average length of stay being 17.5 +/- 3.9 days. Patients were unable to use sipping of enteral feed. Resting energy expenditure neither on admission nor in critical period differed from predicted value. Serum protein concentrations significantly decreased on discharge with normalisation as soon as to day +28, correlating inversely with changes in extracellular water content. Nitrogen urine loss was 10-16 g/day. Only 42.5% of patients were treated with parenteral nutrition. Cumulative nitrogen balance at day +9 was -30.7 +/- 24.1 g N. Body weight at day +28 significantly decreased (-2.94 +/- 4.4 kg), mostly consisting of loss of pure body cell mass. Glutamine supplementation did not improve any of the listed nutritional parameters. CONCLUSIONS: High-dose chemotherapy with autologous stem cell transplantation causes proteocatabolism of medium severity. Nutritional status of patients cannot be improved by the mode and dosage of parenteral glutamine used in our study. Optimal nutritional monitoring and treatment for this group of patients is suggested. PMID- 12577456 TI - [A questionnaire survey of tolerance and adverse effects of clodronate (Bonefos) in patients with multiple myeloma]. AB - The tolerance of Bonefos was assessed by means of a questionnaire completed by 87 (76.3%) of 114 addressed patients with multiple myeloma who use this drug for prolonged periods. Of 54 patients treated by the oral form 50% were quite free from complaints, and complaints associated with more than half the administrations was reported by 7.4% patients. Of 33 patients treated by infusion 57.6% were quite free from complaints and 9.0% patients had complaints after more than half the administrations. The most frequent undesirable effect reported by the addressed patients were digestive complaints. The latter were reported by 47.3% of the total number of patients who had oral treatment and 42.4% patients who had treatment by the i.v. route, although in both groups this undesirable effect was as a rule observed in less than 50% administrations of the drugs. As this was not a placebo controlled trial (which in this case would not be possible for ethical reasons), it is important to consider the fact that the etiological cause of complaints could be any of the concurrently administered drugs incl. cytostatics and that the number of digestive complaints caused by clodronate will be smaller. A total of 16.4% patients with the oral form and 18.2% patients with the intravenous form reported paraesthesias. This emphasizes that it is advisable or rather necessary to ensure calcium and vitamin substitution also in originally normocalcaemic patients. The preference of a 800 mg capsule to two 400 mg capsules was statistically significant. PMID- 12577457 TI - [Diagnosis and therapy of ethylene glycol poisoning--analysis of 20 patients]. AB - The paper presents a retrospective analysis of 20 patients with acute ethylene glycol intoxication who were treated from 1972 to 2001 in the Dialysis Centre of the IVth Medical Clinic and from 1997 at the L. Pasteur Nephrological Clinic of the Faculty Hospital and Safarik Medical Faculty in Kosice. The ethylene glycol intoxication was manifested by neurological symptoms, extreme metabolic acidosis, acute toxic hepatitis and acute renal failure. Laboratory examination revealed oxaluria in 17 patients and leukocytosis in all patients. Percutaneous renal biopsy was made for differential diagnostic or forensic reasons in the convalescent stage of acute renal failure in 6 patients. In the treatment of ethylene glycol intoxication the authors used ethyl alcohol as an antidote: in the first four patients administered by the i.v. route, in the remaining patients in dialyzation solution. Extreme metabolic acidosis improved in 15 patients after bicarbonate haemodialysis. Concomitant application of haemoperfusion over active charcoal during the first haemodialysis in four patients led to a potentiated effect on the uraemic syndrome. Conservative and extracorporeal elimination treatment of ethylene glycol intoxication succeeded in 16 patients (80%). According to the authors' experience early and fractionated application of bicarbonate haemodialysis with 100 mg% concentration of ethylalcohol in the dialysis solution is the method of choice in the treatment of ethylene glycol intoxication. PMID- 12577458 TI - [Thyroid disease in pregnant women and its development after childbirth]. AB - OBJECTIVE: To assess the incidence of post-partum thyropathies in women with positive antibodies against thyroid peroxidase in the second trimester of pregnancy. MATERIAL AND METHODS: Based on a cross-sectional study of the prevalence of abnormal laboratory parameters of thyroid function in a group of 650 pregnant women in the second trimester of pregnancy the authors invited for subsequent clinical examination and repeated controls women with elevated antibodies against thyroid peroxidase (antiTPO) above 60 mU/l (1). The group comprised 75 pregnant women, incl. 44 (58.7%) who attended the examination. After delivery the authors examined the serum levels of TSH, fT4, antiTPO, if necessary fT3 and TRAK. RESULTS: 18, i.e. 58.1% women developed post-partum thyroiditis, incl. 10, i.e. 55.6% who developed hypothyroidism and 8, i.e. 44.4% who developed hyperthyroidism. CONCLUSION: Based on the presented study the authors recommend in pregnant women in the first trimester of pregnancy to introduce assessment of thyrotropin (TSH) and free thyroxin (fT$) and antibodies against thyroid peroxidase (antiTPO)[to detect neuropsychic sequelae on the infant after delivery. The investigation of serum levels of antiTPO antibodies will make it possible to select a group of women in risk of by the development of post-partum thyropathies which must be followed up. PMID- 12577459 TI - [Effect of demographic, anamnestic and clinical factors on hospital mortality in patients with myocardial infarct]. AB - The objective of the work was to analyze local priority data on the possible effect of demographic, anamnestic and clinical factors in a non-selected population of 3123 patients with acute myocardial infarction (AIM) on the hospital mortality (HM). 12.6% patients with AIM died in hospital. The mean age of those who died was 71.92 years. There were more than 4 times more patients above 64 years than 64-year-old ones or younger ones who died. The mortality rate of 64-year-old patients and younger ones (5.2% is significantly lower than in patients above 64 years (19%). The mortality rate of patients above 74 years was 27.1%. Important correlations of the HM were moreover found--in women, patients living permanently in rural areas, in widowed subjects, in patients with elementary education, old age, pensioners and non-smokers. The HM was lower (< 10%) in patients with a history of complex ventricular arrhythmias, impaired lipid metabolism and those who had no other serious disease in the case-history. A higher HM (> 15%) was recorded in patients with a history of a cerebrovascular attack, with data on heart failure and in diabetic patients. The majority of patients (39.4% of all who died) died within 24 hours after admission. During the first three days 57% patients died. Analysis of the characteristic of AIM and HM revealed some clinically important data on the HM less than 30% (patients with complicated AIM, with elevated ST segments, with a left ventricular ejection fraction of less than 40% and with and concurrent heart failure). An adverse course of the disease with a high HM (> 30%) was found in patients with complications of AIM. It was highest, more than 60%, in patients after implemented cardiopulmonary resuscitation, in patients with a combination of three markers of imminent shock and patients in shock. The local priority findings on the HM assembled in Slovakia in a non-selected population of patients with AIM confirm that the high HM still persists in patients of advanced age and in women. It is adversely influenced also by some demographic data, educational level, some anamnestic and clinical factors. The HM of patients with AIM may be adversely influenced also by side-effects of protracted economic transformation which is under way. Data assembled in Slovakia are comparable with similar results assembled in other countries. PMID- 12577460 TI - [Gastropathies caused by non-steroidal antirheumatic agents]. AB - Non-steroid antirheumatics are among very frequently prescribed drugs. They can be, however, associated with the development of serious and undesirable effects on the digestive tract. Therefore at present therapeutic procedures are sought which will restrict the negative effect of drugs on the mucous membranes of the stomach, duodenum and gut in general, incl. new forms of non-steroid antirheumatics with a "sparing" effect on the gastroduodenal mucous membranes. This group of drugs includes preparations with a so-called preferential or specific effect on cyclooxygenase 2 (COX-2). This enzyme is of fundamental importance for the development of mucosal lesions and its inhibition is the basis of safe medication with non-steroid antirheumatics. Other possible alternative therapy is the combined administration of non-steroid antirheumatics with drugs which inhibit the secretion of gastric hydrochloric acid. These drugs include above all proton pump blocking agents. The principle of medication with non steroid antirheumatics should, of course, be careful and correct indication, knowing possible complications of treatment, and in particular knowledge which groups of subjects may be threatened by treatment with non-steroid antirheumatics. Preference of drugs on the basis of COX-2 inhibition is a necessity in these subjects. PMID- 12577461 TI - [Primary hemochromatosis]. AB - Authors describe in the case report a development of the primary hemochromatosis. At the time of the diagnosis in 1979 from organ complication the liver fibrosis and secondary diabetes mellitus were presented. Patient was treated with the venepunctions and with desferoxamine. In the 1998 the primary hepatocellular tumor was found and resected. Instead of the chemotherapy occurred the progression of the disease with multiple metastases and liver insufficiency and 66-years old patient died 19 months after surgery in the 20th year of the disease. PMID- 12577462 TI - [Protection of the heart from reperfusion injury and ineffective oxygen using inhibitors of the mitochondrial permeability transitional pore]. AB - In experiments on isolated hearts of guinea-pig, perfused under Langendorff preparation, possible protection of hearts from reperfusion injury by the known inhibitors of mitochondrial permeability transition pore--cyclosporin A, and trolox--water-soluble vitamin E was studied. It has been shown that cardiac reperfusion was followed with an increase in an oxygen cost of myocardial work (by 83% from control level in 40 min of reperfusion), in addition to the disturbances of cardiac contractility, tone of the coronary vessels and heart rate. The heart function and myocardial oxygen metabolism disturbances, due to global 20 min ischaemia and reperfusion, were essentially decreased by a preliminary application of investigated agents. Trolox improved cardiac recovery both when it was perfused in vitro and after its administration per os. In 40 min of heart reperfusion LVdeveloped pressure was 79% as compared to 51% in that at control; dP/dtmax and dP/dtmin were 88% and 85% accordingly against 66% and 45% in control; oxygen cost of myocardial work didn't change reliably). CONCLUSION: Postreperfusion disturbances of cardiac contractility, tone of the coronary vessels and heart rate, as well as noneffective oxygen utilization by the heart tissue were due to an opening of mitochondrial permeability transition pore. PMID- 12577463 TI - [Change in contractile vascular function in arterial hypertension of different genesis and its correction with phosphatidylcholine liposomes]. AB - We examined the effects of phosphatidylcholine liposomes (PCL) upon the contractile vascular dysfunction in spontaneously hypertensive rats (SHR) and gamma-irradiated (60Co, 6 Gy) rats and rabbits. A significant impairment of endothelium-dependent relaxation was evident in both SHR and irradiated animals. An important novel finding of these experiments is that the impairment was mainly due to the loss of NO-dependent component of relaxation, with the component of relaxation mediated by EDHF being preserved. PCL were found to restore endothelium-dependent relaxation in both SHR and the irradiated vascular tissues. It is important to note that irradiated animals exhibited distinct and sustained signs of hypertension (blood pressure (BP) increased from 122 +/- 8 to 185 +/- 6 mm Hg). Being administered in a single dose of 30 mg/kg, 1 h after irradiation, PCL prevented hypertension development in an early post-irradiated period (9 days). At a later post-irradiated period (6 months), PCL in the single dose lost such a protective effect. Single administration of PCL in SHR led to a transient decrease in BP, but their repeated daily administration caused a persistent decrease in BP up to its normalization as early as in 4 days. These results suggest that PCL possess hypotensive activities due to their ability to normalize endothelial function. PMID- 12577464 TI - [Effects of antibodies on contractility of papillary muscle in the rat heart and duration of descending phase of action potential plateau under influence of exogenic arachidonic acid and inhibiting its metabolism]. AB - The effects of both arachidonic acid (AA) and inhibition of its metabolism on the antibody-induced changes in the mechanical contraction and action potentials of the isolated papillary muscles were studied. It has been shown that antibody dependent induction of AA oxidation led to the initiation of Ca2+ entry through voltage-dependent Ca2+ channels of a myocyte. Inhibition of the lipoxygenase way of AA peroxidation prevented Ca2+ overload of cardiomyocytes more than inhibition of the cyclooxygenase way, but less than blocking Ca2+ channels. PMID- 12577465 TI - [Influence of arginine on the renal functional condition of rats in sublimate nephropathy]. AB - On a background of sublimate nephropathy there was studied the influence of arginin introduction on renal functional condition of white rats. Is established, that the dosage of arginin chloride 300 mg/kg of body weight at in 24 hours after sublimate introduction decreases the kidneys damage and expression of retention azotemia. The positive arginin action in sublimate nephropathy is shown the increasing of glomerular filtration rate and proteinuria decrease. Arginin does not influence on expression of sublimate infringement of kidney Na-transport, and increases it filtrate a charge and renal excretion of present cation. PMID- 12577466 TI - [Effect of irbesartan--angiotensin II type I receptor inhibitor on oxidative metabolism of lipids in essential hypertension]. AB - We evaluated the alters in plasma vasoconstriction eicosanoids (LTC4, TXB2) and diene conjugates (DC) levels in patients with essential hypertension (EH) after chronic (30 days, 235 mg per day) of irbezartane (inhibitor of ANG II type 1 receptor with prolongation action "Aprovel" from "Sanofi") oral administration. Patients with EH have significantly higher plasma both LTC4, TXB2 and DC levels then healthy controls. This imbalance can be beneficially modulated by chronic irbezartane ("Aprovel") administration. It is concluded that ANG II type 1 receptors can be involved in regulation of free arachidomc acid oxidation. PMID- 12577467 TI - [Low conductivity cationic channel in intact endothelium of the rat aorta]. AB - A new low-conductance cationic channel has been recorded with the patch-clamp technique in excised patches of the luminal membrane of intact endothelial cells. The channel had conductance of 5.4 pS and the lifetime of 46 ms. It was equally permeable for Na+ and K+, but impermeable to Cl-. The lifetime and probability of the open state of the channel were independent of the membrane potential. PMID- 12577468 TI - [Effect of kynurenic acid on catecholamine content in the brain of rats with different susceptibility to seizures]. AB - The influence of kynurenic acid on the catecholamine contents in brain of rats with different seizure susceptibility was studied. In animals with high seizure susceptibility the decrease in norepinephrine content and the increase in the coefficient ratio between dopamine and norepinephrine have been found in the most of the regions investigated. Kynurenic acid caused considerable changes in the contents of catecholamines and their precursor in the brain of both groups of animals. The influence of kynurenate on the norepinephrine content was especially significant. The results of a comparison of the absolute contents of catecholamines, as wel las the ratio between the contents of a precursor and the transmitter and the levels of the transmitter in the appropriate nervous terminals are the base to suggest that kynurenic acid enhances the conversion of tyrosine into dopamine, increases its transport to nervous terminals, and decreases the noradrenergic transmission. PMID- 12577469 TI - [Effect of female steroid hormones on expression of adhesion molecules by peripheral blood leukocytes]. AB - The specific adhesion of cells to other cells or to extracellular matrices is a basic component of cell migration and recognition, and it underlies a lot of biologic processes including embryogenesis, tissue repair, and both immune and inflammatory responses. The aim of this study was to investigate the influence of female steroid hormones on expression of the adhesion molecules on the leukocytes. The whole blood from healthy people was incubated in a presence or an absence of progesterone (2 micrograms/ml) or 17 beta-estradiol (0.2 microgram/ml) for 4 h., and then with TNF for 18 h. The phenotype of the leukocytes was investigated by flow cytometry. Progesterone inhibited an expression of CD54 on monocytes and lymphocytes due to reducing density of these molecules on the cellular surface; 17 beta-estradiol inhibited an expression of CD54 on monocytes and CD69 molecules on monocytes and lymphocytes due to reducing density of these molecules on the cellular surface. Progesterone inhibited TNF-stimulated CD54 and CD11b expression on the granulocytes and CD69 expression on lymphocytes by reducing partly the density of these molecules on the surface of cells, and in such way it partly blocked the proinflammatory activity of this cytokine. Progesterone also reduced CD62L expression on the granulocytes by reducing an amount of a marker, positive to those cells but enhanced the effect of TNF. The data obtained evidence that female steroid hormones take part in the regulation of an expression of adhesion molecules by the leukocytes and are likely to be important in the circulation and activation of the leucocytes. PMID- 12577470 TI - [Effect of acupuncture on dynamics of changes in lipid peroxidation and activity of antioxidant system in rats with gastroduodenal ulcer]. AB - In experiments on rats the influence of acupuncture on the lipid peroxidation processes and activities of the antioxidant systems in the gastric mucous membrane, the diencephalon and blood serum at ulcer processes in the gastroduodenal region were studied. It has been determined that acupuncture prevented mainly brain structures from the formation and accumulation of the products of lipid peroxidation that confirmed the neurogumoral influence of acupuncture on the central and peripheral links of ulcer genesis. PMID- 12577471 TI - [Peripheral blood and immunocompetent organs in different adaptational reactions in experiment]. AB - The types of adaptational reactions, masses of the thymus, the spleen and the adrenal glands as well as the thymus histology and morphometry were studied in 79 healthy male rats which received one kind of medicines (sulfalen, pentoxyl, aspirin or ibuprophen), and in 20 animals as a control group. It was established that a quiet activation occurred in 24% of animals, an increased activation--in 22%, an orientation one--in 20%, a defective adaptation--in 23%, stress--in 6%, and an overactivation--in 5% of rats. Changes in the immunocompetent organs, the adrenal glands and the peripheral blood were typical for all types of the nonspecific general adaptational reactions. The data obtained evidence that the adaptive reactions of both a quite and an increased activation should be considered as a contributory factor for body resistance as they are associated with mass enlargement, morphological signs of an activation of the organs of the immune system, and prevailing the mineralocorticoid function of the adrenal gland over the glucocorticoid one. Morphological changes in the thymus in stress reactions, in overtactivation and a defective adaptation are believed to be the background of many pathological processes. PMID- 12577472 TI - [Reactions of lipid peroxidation and haemostasis in various organs in acute emotional-pain stress]. AB - The experiments on rats have shown that an acute emotional-painful stress activates lipid peroxidation and caused hypercoagulation. The role of some organs (brain, heart, kidney, stomach) in those reactions was determined to be different: the brain tissues supplied the blood mainly with thromboplastin, but stomach--with activators of fibrinolysis; brain and heart tissues enhanced the antioxidative properties of the blood, while the stomach tissues reduced them. Such a peculiarity in changes in the lipid peroxidation and haemostasis at the emotional-painful stress should be taken into consideration at an analysis of a pathological process in above organs. PMID- 12577473 TI - [Effect of nifedipine, vitamin E, and bisphosphonates on intensity of lipid peroxidation in arterial and venous walls in hypervitaminosis D]. AB - The peculiarities of lipid peroxydation (LPO) following injection of vitamin D in high doses (10,000 U/kg) and other agents (bisphosphanates, vitamin E, nifedipin) were examined in both arterial and venous walls of rabbits. It has been shown, that tan injection of the bisphosphanates didn't influence significantly the amount of both the intermediate and the final LPO products. Nifedipin did not change the intensity of LPO in the arterial vessels, but it limited the LPO reactions in the venous walls. Vitamin E decreased considerably an amount of the LPO products in all types of the vesselsunder exploration in the animals with hypervitaminosis D. PMID- 12577474 TI - [The role of oxidative metabolism in development of adaptation effect after exposure to ethanol and pulsed hypoxic training]. AB - The activities of lipid peroxidation and the antioxidative defence systems, the content of nitric oxide metabolites, an erythrocyte resistance to peroxide hemolysis in rats exposed to ethanol and hypoxic training were studied. We also evaluated the detoxicant capacity of the liver by 4-aminoantipyrine clearance. It was established that an effect of ethanol was accompanied by the disturbances in the oxygen-dependent metabolic reactions if the liver in different organs and systems. The results have confirmed the expediency of using the hypoxic training in the complex treatment of the chronic alcohol intoxication. PMID- 12577475 TI - [Metabolic effect of amaranth oil and impulse hypoxic training under chronic fluoride intoxication and small doses of ionizing radiation]. AB - Lipid peroxidation and antioxidative defence system in blood, liver and heart tissues, nitric oxide metabolites content in brain tissue of rats under binary action of small-doses of ionizing radiation and fluoride intoxication treated by amaranth oil and interval hypoxic training have been studied. Complex using of amaranth oil and interval hypoxic training result in increase both enzymatic, as nonezymatic links of antioxidant defence in all investigated tissues. It was revealed also enhance of NO system metabolites content in brain gomogenate. In this conditions lipid peroxidation processes in liver and heart tissues normalize comparison with essential increase level LPO under binary action influence. On the basis of obtained results LPO metabolites content we can suppossed that complex using of amaranth oil and interval hypoxic training result in increase of organism adaptative possibility. This complex can be using for binary action of ionizing radiation and fluoride intoxication correction. PMID- 12577476 TI - [Pathophysiologic aspects of endothelial NO-synthase genetic polymorphism]. AB - Information about fourteen allelic variants of promoter, exons and introns of a gene of endothelial NO-synhase (eNOS) dealing with their role in a susceptibility to cardio-vascular diseases has been reviewed. Data of the populational genetic studies, performed in different regions of the world, were analysed to show the interrelation between an availability of on allele in the genome and a risk of ischemic heart disease. The main attention was focussed on the clarification of a relation between some allelic variants of the gene and functional (biochemical) properties of the protein, encoded by this gene, as well as on two principal mechanisms of realization of the pathological allelic variants in eNOS gene: 1) forming the protein in an insufficient quantity or with an altered activity (an interrupted transcription, and stability of informational RNA, formation of a cathalitically deficient protein); 2) an increased protein degradation (due to an acidic hydrolysis or an enhanced proteasomal proteolysis). In practical aspect, the most important problem, according to the point of view of the authors, is that of searching the pharmaco-therapeutical remedies, able to influence the different stages of the molecular-biological realization of an altered eNOS gene (transcription, translation, posttranslational modifications and degradation). Clarification of the above pathogenetic mechanisms will open broad perspectives in constructing therapeutical schemes for individuals possessing the pathological alleles of this gene. PMID- 12577477 TI - [Mechanism of hearing and cochlear physiology]. PMID- 12577478 TI - Mycosis fungoides of the larynx: case report and review of the literature. AB - Mycosis fungoides is a rare cutaneous lymphoma. Dissemination to extra-cutaneous sites occurs at advanced stages of disease. Laryngeal manifestations of mycosis fungoides have been reported in only 13 cases in the available literature. We present a further calla of mycosis fungoides of the larynx at the terminal stage of disease with an additional manifestation in the left maxillary sinus and review the cases published to date. To our knowledge this is the first reported case of mycosis fungoides with laryngeal and paranasal sinus manifestation. Concluding from the present case and from literature therapy should be palliative to improve quality of life. PMID- 12577479 TI - Lipoma of the larynx. Report of 2 cases and review of the literature. AB - Among the primary mesenchymal tumors of the hypopharynx and larynx lipomas are very rare, as they often look macroscopically like retention cysts. Up today approximately 112 cases have been described in literature. We present two further cases of laryngeal lipomas, which were removed endoscopically and by an external approach. Both patients were free of local recurrence. PMID- 12577480 TI - [Correlation of sex hormones in blood serum and estrogen-progesterone receptors concentration in patients with laryngeal cancer]. AB - Particular feature of larynx cancer epidemiology is large difference in morbidity at man and woman. It can be explained not only by exogenic factors, but also by endogenic once, including hormones. There was estimated estradiol and progesterone value in blood serum. The examined group comprised 12 larynx cancer patients. It was used radioimmunological assay Spectria by Orion Diagnostica. At the same time the estrogen and progesterone hormone receptor concentration was estimated in the neoplastic tissue in 18 patients. In the control group, in 10 patients the concentration was examined in surrounding, macroscopically unchanged tissue. The concentration of receptors was estimated by immunoenzymatic assay ER EIA and PgR-EIA by Abbott. The quality assessment was done both in cytoplasmatic and nuclear receptors. There was performed correlation between hormone value in blood and receptor concentration in larynx cancer tissue at 12 patients and in non-neoplastic tissue at 5 patients. There were many abnormal steroid hormones value in blood serum. We found positive correlation between estradiol and progesterone value in blood and concentration of their receptors in larynx cancer. There were observed anomalies of hormone-receptor system at larynx cancer patients. PMID- 12577481 TI - [Problems of differential diagnosis of orbital granulation]. AB - Three cases of orbital tumours caused by granulation are presented. They required advanced differential diagnosis between sarcoidosis, tuberculosis and Wegener's granulomatosis. Each of the patients had an orbitotomy according to Kronlein performed. All were cured. Because of vague results of the histopathological examinations, immunological tests such as: indirect immunofluorescence, direct immunofluorescence and immunodiffusion were performed and cANCA levels were evaluated in order to establish the final diagnoses. PMID- 12577482 TI - [Evaluation of stapes prosthesis biocompatibility]. AB - We reported patient suffering from otosclerosis, who was reoperated after 18 years. During the reoperation we did not observe any changes of intolerance, but histological examination of removed tarflen prosthesis showed foreign body reaction. PMID- 12577483 TI - [Hypothyroidism after combined therapy of laryngeal cancer]. AB - The hormonal function of the thyroid gland in patients with laryngeal cancer, treated with combined method, has been discussed. The first group of them was having one of the thyroid lobes used for reconstruction of the structural defects following partial laryngectomy, the second one, for oncologic reasons--part or the whole gland--excised. Hormonal dysfunction of the thyroid gland was present in 7/10 of the treated patient. In 2 of them the full hypofunction (FH) of the thyroid gland could be observed. Most likely the hypothyreosis was the result of the surgical interference in the thyroid gland (disorders in blood supply in translocated for larynx repair thyroid lobe, excision of the lobe/s for oncological reasons) and degenerative process due to the external irradiation. The monitored hormonal function of the thyroid gland revealed the possibility of transformation of subclinical hypofunction (SH) into FH even after long period of therapy termination. The obligatory monitoring of the thyroid gland hormonal activity for many years has been stressed, as well as the necessity of supplementary therapy at the time of SH. PMID- 12577484 TI - [Lymphocyte subpopulations in middle ear effusions]. AB - The aim of this study was to identify lymphocytes subpopulations in 55 middle ear effusions and peripheral blood samples from 33 children undergoing myringotomy for otitis media with effusion. CD3+, CD4+, CD8+, CD19+ and NK cell populations were investigated using three-colour monoclonal antibody and flow cytometry to quantitative estimation. CD3+ cells were the predominant cell type in effusions (mean 63.3%). Percentage CD4+, CD8+ cells and CD4+/CD8+ ratio was significantly higher in middle ear effusions than in blood. Lymphocyte subset was compared between 22 pairs of effusions from each patient. Percentage of each cells did not differ significantly. The results of this study indicate local regulation of lymphocyte profile in middle ear effusions and the same stadium of immune response in two ears of the same patient. PMID- 12577485 TI - [Personal long-term reproducibility of the TEOAE time-frequency distributions]. AB - The TEOAE signal is relatively short and content changes in time. Due to the fact that signal is very short (20 ms) a special approach must be applied for the analysis of this nonstationarity. Usually the spectral estimation in TEOAE is based on procedures directly employing the fast Fourier transform (FFT). Because TEOAE signal is non stationary the classical Fourier analysis is not adequate for this signal. The aim of our study was the personal comparison of the TEOAE spectograms by using Wigner-Ville distribution (WVD) before and after one year period. Material consisted of 152 men (304 ears) in the age between 18-19 year with good quality OAE. After one year period the comparison of WVD spectrograms showed the highest similarity in the 274 ears (90%), less in the 24 (8%) and in 6 (2%) similarity was not noticed. Personal similarity of the spectrum TEOAE and differences between individual people's ears despite a long time, give a proof about individual architecture of outer hearing cell and maybe useful in the biometrics as a ear-print. PMID- 12577486 TI - [Vestibular system in patients with sensorineural hearing loss]. AB - A near localization of anatomical structures of an inner ear and possibility of damage caused by the same etiopathogenic factor implies that sensorineural hearing loss may be accompanied by vertigo and dizziness. The aim of a study was to evaluate a frequency of vestibular system dysfunction in patients with sensorineural hearing loss and to analyse it according to severity and type of the hearing loss. We evaluated a group of 126 persons which were treated in ENT Department during the last year from sensorineural hearing loss. In all subjects' audiological and ENG examinations were performed. A 50% of patients complained of vertigo and 30% of them--of dizziness. Abnormal ENG recordings were registered in 72% of persons especially in positional and visual ocular-motor tests. In the group without vertigo incorrect ENG tests were often recorded. In the group with vertigo the peripheral and mixed lesions in ENG examination were noted statistically more frequent. We found lack of correlation between magnitude and type of hearing loss and vestibular impairment. Only in the group-residual hearing/deafness were observed significantly more frequent peripheral vestibular dysfunction. PMID- 12577487 TI - [A case of bilaterally elongated styloid processes]. AB - Elongated styloid process gives a complex of symptoms described by Eagle such as unilateral craniofacial and cervical pain, recurrent throat pain, dysphagia and foreign body sensation. Symptoms might be similar to those caused by calcified stylohyoid ligament. Diagnosis of the elongated styloid process is based on the palpation of the tonsillar area and should be confirmed radiologically. Surgical shortening of the elongated process is the only way to give symptomatic relief to the patient. The authors present a surgically treated case of bilaterally elongated styloid processes and unilateral symptoms. PMID- 12577488 TI - [Malignant transformation of giant jugular chemodectoma]. AB - Chemodectoma is a very rare neoplasm classified on the border of the benign and malignant tumours. A case of a large paraganglioma was presented. The method of choice in treatment these tumors should consist of surgery and radiation therapy postoperatively. PMID- 12577489 TI - [Laryngocoele]. AB - Three cases of laryngocoele operated in ENT Department University of Medical Sciences in Poznan by an external approach are presented. Role of larynx tomography in diagnosis procedure and surgical technique is emphasised. Origin, classification, diagnosis and treatment methods are discussed. PMID- 12577490 TI - [Cat scratch disease--diagnostic problems]. AB - Etiology, epidemiology and clinical symptoms of Mollaret's disease, also known as Cat-scratch disease, were described. Four cases of treated patients were presented. One patient with a history of type I Diabetes mellitus presented clinical symptoms of Mollaret's disease, however, after investigation he was finally diagnosed with malignant lymphoma and was given chemotherapy. In other cases some symptoms were also doubtful, but the final diagnoses were made upon positive results of serological reactions and all patients recovered completely after a few week antibiotic treatment. All patients had frequent contacts with cats. A significant divergence between the results of histopathological, serological and ultrasonographic findings lead to a conclusion that a certain diagnosis of Cat-scratch disease can only be made with the use of positive results of serological reactions. PMID- 12577491 TI - [Rare case of laryngeal tuberculosis]. AB - In this work was described one case tuberculosis of the larynx. Provisional diagnosis was tumor of the larynx, because of similar symptoms. Authors described diagnostic and treatment which should be carry out in the similar cases. PMID- 12577492 TI - [Pleomorphic adenoma of minor salivary glands]. AB - Pleomorphic adenoma, a benign tumor often seen in ENT practice, arises either from minor as well from major salivary glands. 5-14% of tumors occur in minor glands. Sixteen cases of minor salivary gland pleomorphic adenomas are studied. Eight of them originate from the oral cavity: 6 from the palate, one from the lower lip and one from the cheek. Two oral adenomas were malignant. In 4 patients tumors occur in the nasal cavity and in two other patients--in the neck. The clinical and pathological features of these patients are presented. Special attention is given to malignant transformation and the rate of recurrence of the tumors. PMID- 12577493 TI - [A case of Kartagener's syndrome]. AB - The authors present a case of a soldier with Kartagener syndrome and bilateral purulent maxillary sinusitis with nasal polyps and pneumonia. Kartagener syndrome is originally described as the combination of situs inversus, bronchiectasis and sinusitis. Kartagener syndrome is a part of immotile cilia syndrome. Mild before the age of 18 recurrent pneumonia, chronic sinusitis, chronic rhinitis and nasal polyps occurred. Typical therapeutic management is discussed but infections prophylaxis, general treatment and physiotherapy are the base of good general condition and significantly improve prognosis. PMID- 12577494 TI - [The achievements of eminent Warsaw physicians in otolaryngology at the turn of the 19th and 20th century]. AB - The scientific achievements of many Warsaw otolaryngologists in the second part of the 19th and in the beginning of the 20th century are presented. Karol Benni's (1843-1916) contribution to the world otology is described. The famous Warsaw laryngologist Teodor Heryng (1847-1925) was a creator of a new operating method of the larynx tuberculosis by curettage. He was also a propagator of the diaphanoscopy (transillumination) of maxillary sinuses. Ludwik Guranowski (1853 1926). a distinguished otiatrist performed a first myringoplasty in the Polish territories and explained the causes of external otitis by bacillus pyocyaneous. Jan Sedziak's (1861-1932) work on malignant neoplasms of larynx was given a prize at international congress in Tuluse in 1893. Zdzislaw Dmochowski (1864-1924) described the pathologic anatomy changes in maxillary sinuses. Benni and Alfred Sokolowski (1849-1924) were the authors of several chapters in foreign medical textbooks. Heryng's inhalation apparatuses exhibited in Paris are described. Sedziak, Przemyslaw Pieniazek (1850-1916) and Teodor Heiman (1848-1917) were also the authors of articles in foreign otolaryngological journals written in connection with the jubilee of famous otolaryngologists: Leopold von Schroetter, Bernhard Fraenkel and Hermann Schwartze. The problem of focal infection in otolaryngology described by Feliks Erbrich (1874-1938) is briefly pointed out. PMID- 12577495 TI - [The report from the Fifth International Conference on Otorhinolaryngologic Diseases in Pediatrics, Graz, July 8-11, 2002]. PMID- 12577496 TI - [The report form the XXVI International Audiology Congress, Melbourne, Victoria (Australia), March 17-23, 2002]. PMID- 12577497 TI - [Pregnancy complications with abnormal results of biochemical screening for Down syndrome in second trimester and normal fetal karyotype]. AB - Pregnancy complications were studied in the following groups of pregnancies with structurally and chromosomally normal fetuses: 1) 216--with elevated free beta hCG above 2 MoM and 134--above 2.5 MoM; 2) 37--with elevated alpha-fetoprotein [AFP] above 2 MoM; 3) 67 screen-positive patients below the age of 35. The following complications were compared in the studied and the control groups: preterm delivery and premature rupture of membranes [PROM], preeclampsia (in general and severe one), IUGR without preeclampsia, delivery of small for gestational age [SGA] and low birth weight [LBW] infants and perinatal fetal loss. Pregnancy outcome in cases with free beta-hCG above 2 MoM was not different from that in the control group. With 2.5 cut-off level the incidence of preeclampsia and of LBW infants was higher in the studied than in the control group. Elevated AFP above 2 MoM alone was associated with more frequent preeclampsia including severe one (in all the latter cases--IUGR present). In the screen-positive group the incidence of preeclampsia (in general and severe one with IUGR), SGA and LBW infants and perinatal mortality rate were significantly higher than in the control group. No difference was found between the studied and the control groups regarding subsequent development of IUGR without preeclampsia, preterm delivery and PROM. Abnormal second trimester serum screening test results (elevated free beta-hCG or AFP alone above 2.5 and 2 MoM respectively) in pregnancies with normal fetal karyotype indicate higher risk for subsequent development of preeclampsia. Screen-positive patients, being a heterogeneous group (including ones with elevated free bb-hCG alone and ones with elevated both free beta-hCG and AFP), are at increased risk for other pregnancy complications too. PMID- 12577498 TI - [The activity of enzymes and free radical oxidation in acute and chronic fetal hypoxia in general anesthesia during cesarean section]. AB - The following study dwells on a basic problem in obstetrics anaesthesia. It shows a present view of the in NLA and ataractics having effect on free radical oxygenation process in mother-fetus system in Cesarean section. 101 pregnant women have been studied as the values of FROq serum antioxygenase activityq serum enzymes activity in women blood and in blood of umbilical cord have been compared. PMID- 12577499 TI - [Pros and cons of magnesium sulfate as a tocolytic]. AB - Magnesium sulphate (MgSO4) is with a proved effectiveness in cases of praeclampsia/eclampsia, but its use as a tocolytic is discussed in the last few years. That is why we had for an object to study its effect as a tocolytic in cases of abortions and premature labours. The study is prospective and if is made in I-st obstetric clinic of High Medical School--Pleven. Treatment with Cormagnesin was carried out to pregnant women for suppressing the uterine activity. Cormagnesin 200 (1 amp-10 ml) contains 1000 mg MgSO4, and Cormagnesin 400 (1 amp-10 ml) contains 2000 mg MgSO4. The medicine was administered in dosage of 4 or 5 g for 30 min, and after that if there were any uterine contractions the infusion was carried on with additional 5 g MgSO4 for 6 to 12 h. The total dosage was from 4 to 60 g MgSO4. The authors reported on very good effect in cases with pains and increased uterine tone--18 (36.73%), as well as in cases with pains and irregular uterine contractions 5 (10.21%), while the treatment was without any effect in cases with uterine contractions on 15-20 min, increased uterine tone, bleeding and Pelvic score 1-3 points in spite of high dosages of MgSO4 and longer duration of treatment. The authors made the conclusion, that the subjective complaints should not be accepted as an indication for administration of MgSO4, and MgSO4 should be administered in cases with increased uterine tone and irregular uterine contractions. Every genital bleeding and suspicion for placental abruption should be defined more precisely, because lately diagnosed placental abruption and unjustified expectation for suppression of uterine activity by MgSO4, may lead to increase of perinatal morbidity and mortality. In spite of the controversial data about MgSO4 as a tocolytic, its administration is justified and necessary. PMID- 12577500 TI - [Candidiasis and birth canal injuries]. AB - Candidiasis of the lower female genital tract during pregnancy is a leading cause for seeking medical help. During pregnancy candidiasis is encountered about 30% more often compared to non-pregnant (by the time of delivery about 18-22% of the pregnant women would have the infection). It is clinically manifested especially after 36 w.g. This might be the cause of a series of pregnancy complications. Most serious are considered prematurity, chorioamnionitis, descending infection from mother to the newborn during delivery. Significantly less attention has been paid to the birth canal injury and its higher rate if a Candida colpitis is present at the time of delivery. PMID- 12577501 TI - [Association between Mycoplasma genitalium and preterm labor]. AB - M. genitalium was identified by using polymerase chain reaction (PCR). The samples were obtained from the posterior vaginal fornix between 21-25 gestation weeks and investigated for PH, Gram Stain for bacterial vaginosis; Chlamidia trachomartis; Neisseria gonorrhoeae; Trichomonas vaginalis. Of 102 pregnant women, only five had a PCR-positive for M. genitalium. The occurrence of M. genitalium in posterior vaginal fornix at midtrimester is infrequent in pregnant women and unlikely to be a contributing factor for spontaneous preterm birth. PMID- 12577502 TI - [The probiotic Lactobacillus acidophilus--an alternative treatment of bacterial vaginosis]. AB - The probiotics or biotherapeutical agents are microorganisms that have an antagonistic activity regarding pathogens in vivo. Therapeutic application of these microorganisms is mostly in cases of infections of non-sterile mucous surfaces--vagina and large intestines. With the help of the probiotic L. acidophilus bacterial vaginosis can be treated successfully in a natural way. The lack of systemic side effects makes it a drug of choice in the treatment of pregnant women. More studies in this direction and development of new strategies for treatment of bacterial vaginosis in pregnant and non-pregnant women are needed. PMID- 12577503 TI - [Paraaortic lymphadenectomy in gynecological tumors]. AB - The purpose of this Study was to determine role of para-aortic lymphadenectomy in patients with gynecologic malignancies. In review literature the authors reports was to show role of paraaortic lymphadenectomy in surgical staging and the most important role in treatment plan. The results confirm the diagnostic and prognostic value of paraaortic lymphadenectomy in patients with endometrial carcinoma, cervical cancer and ovarian cancer and adjuvant treatment after primary surgical procedure. The authors affirm that lymph nodes metastases represent the most reliable markers of high risk patients. The position about therapeutic role of paraaortic lymphadenectomy remains controversial. PMID- 12577504 TI - [Osteoporosis in pregnancy and lactation]. AB - In the second half of the twentieth century different patterns of bone mass changes in young women were described in association with pregnancy and lactation. Pregnancy-associated osteoporosis may present as predominant bone loss from the hip; there is a second form with vertebral involvement; and it may present with fractures at atypical sites such as the sacrum. Long-term lactation is also associated with increased bone loss. In our work we describe a clinical case of a 34-year old lactating woman, presenting at the fourth month after unremarkable childbearing with vertebral fractures and overt osteoporosis. We described the diagnostic procedures and therapeutic interventions. A literature review is also presented. In conclusion, osteoporosis of pregnancy and lactation should not stay out of the clinician's focus and requires a multi-disciplinary approach. PMID- 12577505 TI - [Effect of some factors on obstetrical care of women with previous cesarean section]. AB - Considering obstetrical management we should have always in mind advantages and disadvantages of vaginal birth after cesarean (VBAC) and elective cesarean section after a previous one, financial part of the elected mode of delivery in the particular case, psychological aspect of the matter (especially after an unsuccessful trial of labour--TOL) and not on the last position-patient's right to choose the way how to deliver. We performed an inquiry of 3-year-period and 50 women with a prior cesarean section, by which we studied the factors contributed in different extent to the choice of the following approach to delivery: patient's preliminary choice, the impact of medical staff during pregnancy and when entering the Department of Obstetrics, the impact of non-medical persons, complications after the previous cesarean section, patient's acquaintance with complications and consequences associated with the operative mode of delivery, as well as doctor's personal attitude to the different modes of delivery. The aim of our research is to show that by careful selection (according to the established indices) of women, who have a high probability (more than 80%) to deliver their babies vaginally and offering adequate information by educational literature and including the woman in the decision making process, the patient resistance to vaginal delivery will be reduced. In this way, the growing rate of cesarean section will be decreased by increasing the number of VBAC--undoubtedly one of the most important issue in modern obstetrics. PMID- 12577506 TI - [Treatment of precancerous changes of the uterine cervix]. AB - The aim of the present review is to view the possibilities for the treatment of the precancer lesions of the uterine cervix, on the basis of the modern tendencies in the gynecological practice. The description and criteria for the applicability of the actively observation, different destructive--chemical, thermo-, cryo-, electro- and laser ablation, as well as excisional methods- classical, surgical, laser and LEEP/LLETZ conization were discussed. The advantages and disadvantages of each method were pointed out. The conclusion is, that the presence of different therapeutic strategies enrich the treatment opportunities, which depends, from one hand of the technical equipment, the experience of the specialists working in this field, and the correct indications of the cases from other. PMID- 12577507 TI - [Conservative treatment of ectopic pregnancy with methotrexate]. PMID- 12577508 TI - Information needs of young scientific workers and pre-graduated students in the sphere of pharmacy. AB - The development of the Pharmaceutical science and its information insurance in the conditions of a transition period attain greater importance. The increase of the stream of information is more and more difficult for processing and from there-for usage. PMID- 12577509 TI - In vitro study on capsule formulations of omeprazole containing enteric coated granules. AB - Omeprazole is a unique and effective agent for suppression of gastric acid secretion. It promises to be especially useful in Zollinger-Ellison Syndrome (1) and in other patients who have failed to respond to H2 antagonist therapy. The dose requirement for zollinger-ellison syndrome (ZES) disease is upto 180 mg per day. The drug omeprazole shows biological half life of 2-3 hours. (2) Considering its proven therapeutic efficacy for ZES disease, this study is attempted to formulate sustained release capsule containing enteric coated granules. For formulating sustained release drug delivery system immediate release dose (IR) is calculated as 10 mg of drug and sustained release dose (SR) is taken as 50 mg. (3) The polymers used for retarding the release are Ethyl Cellulose (EC) and Hydroxy Propyl Methyl Cellulose (HPMC). Solvent evaporation technique is followed to prepare sustained release granules. Granules are finally coated with enteric coating of Suteric (Polyvinly acetate phthalate, PVAP). Thus the eight capsule formulations are prepared and formulation found very close to theoretical sustained release (TSR) according to in vitro study (4) is subjected to stability studies. Results show that product should be stored at room temperature or below ambient temperature. PMID- 12577510 TI - Formulation studies on modified releasing amoxicillin trihydrate microcapsules. AB - In vitro researches and in vivo animal experiments show that, generally the antibiotic concentration should be above minimum inhibitory concentration for optimal efficiency. There is also direct and indirect evidence from human trials to support this claim. Consequently, a modified-release dosage form of the beta lactam antibiotics might be therapeutically more efficient than the existing conventional products, which are rapidly absorbed to produce transient peaks in serum drug levels. In our study, Amoxicillin was chosen as the model drug, because it is currently one of the most widely prescribed oral semisynthetic penicillins. By using various types of polymers and different formulations of them, we tried to improve a microparticular system. We succeeded in obtaining optimal production of particulars in the prepared microcapsule form with the solvent evaporation technique. By applying suitable pharmaceutical technological production parameters, the production of the dosage form which releases modified release form of the model antibiotic is provided. Production processes and availability for the aim is controlled by using the pharmaceutical technological tests, mainly with the methods according to USP XXIV. Consequently compared to the classical Amoxicillin trihydrate dosage forms used in treatment, we managed to form the microcapsule structure containing modified release. PMID- 12577511 TI - Synthesis and structural study of substituted thioxothiazolidinones and thioxoimidazolidinones. AB - A new set of derivative thioxothiazolidinones and thioxoimidazolidinones 3,5 dissubstituted has been synthesized with satisfactory yield from the condensation Knoevenagel type between benzaldehydes and 4-thioxothiazolidin-2-one, 2 thioxothiazolidin-4-one and 1-methyl-2-thioxoimidazolidin-4-one compounds following by N-alkylation with aryl or acyl halides. The physico-chemical properties of the 5-benzylidene-3-[2-(4-chlorophenyl)-2-oxoethyl]-2 (or 4) thioxothiazolidin-4 (or 2)-ones and 5-benzylidene-1-methyl-2-thioxoimidazolidin-4 ones synthesized have been described. PMID- 12577512 TI - Photosynthesis of furocoumarin- and furochromone-types potential intercalative alkylating and oxidizing agents of DNA through photooxidations using gamma-ray. AB - The photooxygenation of imperatorin (1a) under gamma-ray irradiation afforded the hydroperoxides 2a and 3a. Similarly, the photooxygenation of alloimperatorin (1b) gave the hydroperoxide (2b). Visnagin (1c) was also photooxygenated to give the hydroperoxide (2c) as sole product. On the other hand, the photooxygenation of khellin (1d) gave the endoperoxide (2d) as a sole product. The epoxidation of imperatorin (1a) using hydrogen peroxide under gamma-ray irradiation afforded the epoxide 5a. Similarly visnagin (1c) and khellin (1d) were epoxidized to give the epoxides 5c and 5d. PMID- 12577513 TI - Synthesis of some new azoles and azines and their condensed derivatives. AB - Ethyl (2,5-Dimethylphenyl) pyruvate (2) condensed with o-phenylenediamine, ethylenediamine, semicarbazide and thiosemicarbazide and gave quinoxaline derivative (3), dihydropyrazine derivative (9) and 1,2,4-triazine derivatives (10, 11) respectively. Compound (3) reacted with phosphorous pentasulphide, polyphosphoric acid, semicarbazide hydrochloride and phenylisothiocyanate yielded thienoquinoxaline derivative (4), furoquinoxaline derivative (5), semicarbazone derivative (6) and pyrrole derivative (15) respectively. Reaction of (2) with hydrazines and malononitrile afforded the arylpyrazole derivatives (12a, b) and pyrane derivative (13) respectively. Heterocyclization of compound (6) afforded thiadiazole and selenodiazole derivatives (7), (8). PMID- 12577514 TI - A facile synthesis of 2-aryl-4,5-di(2-thienyl)imidazoles under microwave irradiation and their antimicrobial activities. AB - In this study, some 2-aryl-4,5-dithienylimidazoles were synthesized by reacting di-(2-thienyl)ethandione and a suitable aromatic aldehyde in the presence of ammonium acetate in acetic acid. Either the classical reflux method or microwave irradiation method were applied as alternative reaction conditions. The antimicrobial activities of the compounds obtained were investigated in vitro and appreciable activities were obtained. PMID- 12577515 TI - Preparation, characterization & anticancer evaluation of potassium N-(p-anisole) alpha-(2-xanthatophenyl)nitrone. AB - The new compound, Potassium-N-(p-anisole)-alpha-(2-xanthato-phenyl)nitrone of the formula K(p-MeOC6H4)N(O) ? CH(O-C6H4CS2) was prepared and characterized by elemental analysis, infrared, UV spectra and 1HNMR spectroscopy. The anticancer activity of it has been achieved by the National Cancer Institute (NCI) U.S.A. A remarkable activity was exhibited against nine types of cancer cells specially the colon, ovarian and CNS cancer. PMID- 12577516 TI - Incubation time in sterility tests for pharmaceutical products. AB - Throughout the evolution of sterility test methodology, different periods of incubation were adopted depending on both the type of microorganism to be detected and the employed inoculation methodology. Some official compendiums, such as Brazilian and Mexican pharmacopoeias, recommend different incubation periods, according to the inoculation methodology employed for the sterility test, whereas the United States Pharmacopeia, in its last edition, started to adopt an incubation period of 14 days, independently from the methodology employed. The aim of this work was the evaluation of the influence of the incubation time on the efficiency of different methodologies for sterility test, as well as the benefits that could be achieved with the incubation time extension. The experiments led to the conclusion that an incubation period of 14 days is enough for the detection of microbiological contaminants in pharmaceutical products submitted to sterility tests, independently from the methodology employed, what values the alteration introduced in the USP XXIV, to the detriment of other pharmacopoeias which maintain different periods of time in accordance with the inoculation method employed. PMID- 12577517 TI - Anti-PLA2 action test of Casearia sylvestris Sw. AB - Casearia sylvestris (Flacourtiaceae) is a plant which grows in the wild. The crude extract and pure substances from this plant induced partial inhibition of the PLA: (phospholipase A2) activity of snake venoms and some purified toxins. C. sylvestris extract efficiently neutralized the hemorrhagic and myotoxic activities caused by crude venoms and toxins. PMID- 12577518 TI - Novel 4-aminopyrimido[4,5-b]quinoline derivatives as potential antimicrobial agents. AB - Two series of 4-aminopyrimido[4,5-b]quinoline derivatives substituted in the 2 position 6a-c and/or in 1-position 9a-e have been prepared by facile routes starting from 2-amino-3-cyanoquinoline 2,2-chloro-3-cyanoquinoline 4 and 2 arylamino-3-cyanoquinolines 8a-d. The reactions involved simple fusion with thiourea or urea and, in some cases, with guanidine. The prepared compounds were in vitro tested for antimicrobial activities against some selected Gram-positive, Gram-negative bacteria and fungi. The products containing the thio-function were the most active followed by those containing the imino-function while the carbonyl containing derivatives were without significant antimicrobial effect. PMID- 12577519 TI - Effect of different gamma radiation doses on solid crystalline insulin: hypoglycaemia in diabetic beagle dogs. AB - The effect of different doses of gamma radiation (25, 50 and 100 KGy) on the solid crystalline insulin was studied. The results showed that there was no significant difference (P > 0.05) between the hypoglycaemic effects of 25 kGy gamma-irradiated and the non-irradiated insulin as indicated by the plasma glucose levels and the relative hypoglycaemia. The exposure of insulin to higher doses of radiations (50 and 100 kGy) resulted in almost the same hypoglycaemic effect, which was however greater than that produced by the 25 kGy gamma irradiated and the non-irradiated insulin. The DSC thermograms showed that the thermal behaviour of crystalline insulin was not affected by the different doses of irradiation. However, a slight colour change was noticed on samples exposed to 50 and 100 kGy of radiation but not on samples subjected to 25 KGy. In conclusion, while the routinely used dose (25 kGy) of gamma radiation for sterilization does not seem to affect the activity of insulin, higher doses of radiation (50 and 100 kGy) enhance its hypoglycaemic effect. PMID- 12577521 TI - [Bioethical aspects in odontostomatology]. PMID- 12577520 TI - Sedative effects of the methanolic leaf extract of Newbouldia laevis in mice and rats. AB - The effect of the methanolic extract of Newbouldia laevis seem on the central nervous system of rats and mice was investigated. The extract was tested on spontaneous motor activity, exploratory behaviour, apomorphine induced climbing behaviour in mice and pentobarbital induced hypnosis in rats. The extract caused considerable reductions of exploratory activity, spontaneous motor activity and prolonged pentobarbitone-induced hypnosis in rats. It was also found to attenuate apomorphine climbing in mice. The results suggest that the methanolic extract of Newbouldia laevis may contain principles that have sedative properties. PMID- 12577522 TI - Survival to oral cancer. A study of clinical risk markers with independent prognostic value. AB - BACKGROUND: The aims of this study have been to describe survival to oral cancer and to identify clinical variables with independent influence on its prognosis before treatment. METHODS: 94 oral cancer patients treated during 1991-99 entered the study. The variables considered were: age, sex, location of the lesion, clinical presentation, symptoms, TNM classification, months elapsed since treatment and ploidy pattern. A descriptive study of the data was performed, along with a survival analysis using Kaplan-Meier curves (log rank test for comparison among curves) and single and multivariate Cox regression. RESULTS: Multivariate analysis recognized a prognostic value for the age of the patient (OR = 1.06; CI95%: 1.02-1.09) and also for tumour size. Tumour stage resulted also selected, but its predictive value was lower than size's, so it was excluded from the predictive model. No statistically significant differences in terms of survival were identified on the rest of variables considered in the study. CONCLUSIONS: This study suggests the need for considering age and tumour size as the most relevant clinical variables for predicting survival to oral cancer at the time of diagnosis. PMID- 12577523 TI - Condylar-mandibular asymmetry, a reality. AB - The aim of this study is to evaluate the possibility to recognize a condylar mandibular asymmetry through a panoramic radiograph. Results from a previous work, in which 100 skulls from the Museum of the Institute of Anatomy of the University of Pavia were studied and measured, showed the presence of asymmetry. Using the same skulls we examined the possible correlation between morphological and radiological data. We did not find out correlation's between the condylar asymmetry evaluated at the anatomical level and the radiological asymmetry which was indeed found. This is probably due to the different positioning of the jaws during the two different measuring processes. Nevertheless our results confirm the daily experience of dentistry: asymmetry of mandibular condyle can be one of the mayor causes for the asymmetry of the stomatognatic apparatus. PMID- 12577524 TI - Expression and modification of NO synthase in human dental pulps during orthodontic treatment. AB - In the last few years our histology laboratory has worked in collaboration with Winchmore Hill Dental Practice in London in studying nitric oxide synthase (NOS) protein expression by the use of immunohistochemistry in dental pulps following orthodontic treatment (A. Gerbino et al. 2000; A. Gerbino et al 2001; A. Gerbino et al 2001.) The study has been carried out on samples taken from orthodontically treated and non-orthodontically treated teeth. The results suggest a close correlation between the duration of the orthodontic traction and the expression of the above-mentioned neurotransmitter (NO). PMID- 12577525 TI - [The persistence of ontogenic characteristics in the adult masseter muscle]. AB - During embryonic and foetal development, the masseter is formed from two successive generations of muscle fibers in a manner which is very similar to that which has been previously described for other skeletal muscles. This phenotype is characterised by the persistence of ontogenic myosin isoforms (embryonic and foetal myosin heavy chains, embryonic light chain) and by the presence of two distinct populations of fibers: small diameter fibers which coexpress the embryonic, foetal and fast isoforms of the myosin heavy chains but never express the slow isoform; large diameter fibers which express the slow myosin heavy chain either exclusively or in variable associations with the other isoforms. These characteristics of the human masseter muscle probably correspond not only to its embryological origin and its special innervation, but also to the functional constraints to which it is submitted after birth. PMID- 12577526 TI - Getting physical. PMID- 12577527 TI - Farewell to 'Aunt Flo'. PMID- 12577529 TI - Herbal stress buster? PMID- 12577528 TI - 'The perfect crime'. PMID- 12577530 TI - Facing bullies. PMID- 12577531 TI - Federation, AHA, AAMC unveil landmark quality reporting initiative. PMID- 12577532 TI - GOP to control Congress, Frist named majority leader. PMID- 12577533 TI - A good beginning. PMID- 12577534 TI - [Views of a surgeon on publishing]. PMID- 12577535 TI - [Review of deaths in patients hospitalized at the Orthopedic-Traumatology Clinic of the 3rd Medical Faculty and at the Kralovske Vinohrady Faculty Hospital in Prague from 1998 to 2000]. AB - In recent years there are in particular in the daily press discussions on the publication of data concerning the number of deaths in different hospitals. Without comments on the structure of the group of hospitalized patients and their morbidity these data may be confounding. The objective of the work is to inform on the number of deaths and spectrum of the deceased at the Orthopaedic Traumatological Department in course of three years and analyze the data. The investigated group was formed by patients who died in the course of hospitalization at the Orthopaedic-Traumatological Clinic of the Third Medical Faculty Charles University and Vinohrady Faculty Hospital Prague in 1998-2000. In 1998 16 men and 15 women died (mean age 85 years), in 1990 29 women and 10 men (mean age 81 years) and in 2000 14 women and 6 men (mean age 78 years). Patients after injuries predominated, only 5 patients died after a planned orthopaedic operation during this period. The most frequent diagnosis of the deceased patients was fracture of the proximal femur which in 1998 and 1999 accounted for more than 90% and in 2000 for 100% of the patients who died after an injury. According to post-mortem diagnoses death caused by acute myocardial infarction and other complications of ischaemic heart disease predominated (55%). PMID- 12577537 TI - [Laparoscopic surgery of gastric diverticula with secondary adhesiolysis]. AB - The authors present the case-history of a rare cause of gastric diverticulum. On account of a very painful symptomatology and X-ray evidence of a diverticulum the authors decided on laparoscopic excision of the diverticulum. The resection was made using a Endo GIA II stapler. The gradual, one-year development of adhesions and a new different algic symptomatology which impeded the subject's occupational activities led the authors to make another laparoscopic operation. After releasing the adhesion between the stomach and the collateral peritoneum the condition improved completely. The patient is now 16 months after re-operation completely free of complaints and performs his physically pretentious work and ingests food without dyspeptic phenomena. PMID- 12577536 TI - [Hand-assisted laparoscopic surgery of the colorectum]. AB - Hand-assisted laparoscopic surgery extends the possibilities of laparoscopic operations while preserving the miniinvasive character of the operation. The objective of the work is, based on the authors' experience with a group of 28 patients operated by this method for diseases of the colorectum to describe the surgical technique, present their results and emphasize the indications and advantages of this procedure. PMID- 12577538 TI - [Entrapment syndromes of the peripheral nerves of the upper extremities]. AB - BACKGROUND: The author presents the results of surgery of entrapment syndromes of the peripheral nerves implemented at his department in the course of 10 years (1990-2000). MATERIAL AND METHODS: In 72 patients 90 operations were performed in the course of which 85 entrapment syndromes were treated. The carpal tunnel syndrome was recorded in 40 patients (55.6%), the cubital canal syndrome in 24 patients (33.3%). The other entrapment syndromes of the peripheral nerves of the upper extremities-8(11.1%) were classified as rare. In 13 patients the entrapment syndrome was bilateral, the carpal tunnel syndrome in 12 and the cubital canal syndrome in one patient. In 17 patients there were re-operations, incl. 12 re operations after primary treatment in another department. The clinical picture of compressive syndrome was divided according to the intensity according to Dellon's classification into 3 grades. Grade I compression was found in 2 cases (2.8%) (2/2). Grade II compression was found in 8 patients (11.1%) (9/10). Grade III Patients with grade III compression-62 (86.1%) predominated (68/80). The values are expressed as the number of patients/number of entrapment syndromes. The effectiveness of the surgical operation was analyzed with regard to the degree of compression, length of compression and duration of symptoms. RESULTS: Excellent and satisfactory results were recorded in all cases with grade I and II compression and duration of symptoms up to 12 months. The position was worse in patients with grade III compression where excellent and satisfactory results were recorded in 62 cases (77.5%) and when the complaints persisted for more than 12 months, i.e. in 30 of 48 cases (62.5%). CONCLUSION: The decisive factor which influences the result of the operation is the grade of nerve compression and its duration. Decisive factors improving the results of surgery are age under 30 years and period of complaints shorter than one year, when the symptoms are not yet irreversible. PMID- 12577539 TI - [Major vascular injuries in laparoscopic operations]. AB - The authors present three case-histories of major vascular injuries during laparoscopic operations. Subsequently they discuss causes and possible prevention of these rare but serious complications. PMID- 12577540 TI - [Prevention of infectious complications by reducing prophylactic administration of antibiotics and observing the hygienic and epidemiologic policies of the intensive care unit]. AB - OBJECTIVE: The authors investigated in a prospective one-year study the incidence of nosocomial infections in patients hospitalized at the intensive care unit with reduced prophylactic administration of antibiotics and adherence to the hygienic epidemiological regime. METHOD: At the intensive care unit of the Neurocentre of the Liberec Hospital nosocomial infection were investigated during the prophylactic administration of antibiotics, most frequently in one dose for a maximum of 24-48 hours. For therapy antibiotics were indicated in proved or suspected bacterial infection. They were not administered in case of colonization of the patient. Concurrently the hygienic and epidemiological regime was ensured. It involved adherence to cleanliness, disinfection, in particular hygienic disinfection of the hands, sterilization, barrier nursing techniques, single-use equipment, closed systems, enhanced regime in case of colonization of the patient with resistant strains and introduction of standard procedures. RESULTS: The one year prospective study comprised 529 patients hospitalized at the intensive care unit at the age of 9 to 85 years with a mean hospitalization period of 4.9 days. 57.1% patients had cerebral diseases, 31.9% diseases of the vertebral column, and other diagnoses accounted for 11%. A total of 78% patients were operated. Antibiotics were administered prophylactically to 76% patients. In the majority this was in conjunction with the surgical operation (98%). To 93% patients the antibiotic was administered only in one dose, only to 3% patients it was administered for 24 hours and to 4% patients for 48 hours. For therapy antibiotics were administered to 10% patients, including 2% patients where treatment was started at another department. Nosocomial infections were recorded in 5.4% patients. Most frequently infections at the site of operation were involved (2.5%) and infections of the urinary pathways (2.1%). Infections of the airways were recorded in 0.8% patients and infections of vascular catheters were not recorded at all. CONCLUSIONS: Adherence to the hygienic and epidemiological regime and rational administration of antibiotics leads at intensive care units to a reduced incidence of nosocomial infection and thus to reduced financial costs. PMID- 12577541 TI - [Initial experience with transplantation of a kidney from a non-beating heart donor (NHBD)]. AB - The authors present their initial experience with transplantations of the kidneys from a donor with a non-beating heart (NHBD). This programme was developed because of the permanently declining number of cadaverous kidney donors with a beating heart. Since January 2002 the authors collected a total of three kidneys from NHBD 3 kidneys of which were used for transplantation. The first two started to function immediately, the third one restored its function three weeks after transplantation. The viability and suitability of use of kidneys from NHBD is evaluated with regard to vascular resistance and flow of the perfusion fluid through the kidneys when using a perfusion pump and by means of totalglutathione S-transferase (tGST) assessed at intervals in the perfusion fluid. The values of the above parameters are limiting between the 6th and 8th hour after the onset of instrumental perfusion of the collected kidneys when the perfusion values should be within the range of 30-50 ml/min/100 g and the vascular resistance should be < 0.7 mmHg and tGST < 100 U/l. For immunosuppression the authors use during the fist days after transplantation basiliximab, rapamycin and corticoids, later in case of satisfactory function of the graft they add cyclosporin A. The initial results of renal transplantation from NHBD are promising and the programme will be developed further. PMID- 12577542 TI - [Respiratory insufficiency in the surgical intensive care unit]. AB - Postoperative pulmonary complications are of clinical importance: they prolong the hospitalization, increase the costs of treatment and contribute to the perioperative mortality. Respiratory insufficiency--inability to maintain adequate exchange of gases in the lungs--is its most severe form. Early diagnostics is essential and it is possible only during continuous and meticulous surveillance of the patient. In the postoperative period there are factors present enhancing the development of the postoperative pulmonary complications: drugs, pain, influence of the trauma of the operation, decreased lung capacity and decreased mobility. Prevention of complications is essential. Its indispensable part is adequate preoperative preparation, as well as good management of anaesthesia. The cornerstones of adequate postoperative care are oxygen therapy, sufficient analgesia, physiotherapy. At the earliest signs of respiratory insufficiency the commencement of respiratory support is mandatory. Artificial ventilation can be used as a preemptive measure preventing the development of pulmonary complications. PMID- 12577543 TI - [Nikolai Il'ich Blinov (1899-1971)]. PMID- 12577544 TI - [Ganglionic blockers in abdominal surgery: facts and hypotheses. I. Benzohexonium and motor function of the gastrointestinal system in the early postoperative period]. AB - The method of registration of the intraluminal pressure was used to study the function of the stomach, small intestine and sigmoid colon under conditions of partial ganglionic blockade in the first days after truncal vagotomy and resection of the stomach. It was found that benzohexonium in doses 0.1-0.3 mg/kg failed to substantially decrease the frequency of early functional motor evacuation disorders of the "operated" stomach, but the results of using N cholinolytic was better after truncal vagotomy than after resection of the stomach. Benzohexonium in doses 0.1-0.2 mg/kg failed to considerably stimulate the motor function of the small intestine while the doses of 0.3-0.4 mg/kg resulted in a decrease of its contractile activity. No reliable changes in the qualitative and quantitative parameters of the sigmoid colon motor function were found against the background of ganglionic blockade. So, for prevention and correction of early postoperative motor-evacuation disorders of the gastrointestinal tract the ganglionic blockade with N-cholinolytics should not be taken as a method of choice or a variant of monotherapy. PMID- 12577545 TI - [Gastroduodenal ulcer hemorrhage under conditions of Extreme North (Yakut)]. AB - Under analysis there were results of treatment of 431 patients with duodenal ulcer bleedings. The authors note that the duodenal ulcer bleedings under conditions of Extreme North (Yakutia) are observed more often in those who come (not in residents). Among specific features of the clinical course of the ulcer disease the authors note a short case history, continuous and severe exacerbation, short remission, frequent development of complications. The authors use in their work active individualized surgical methods and make organ-saving operations aimed at saving the patient's life. PMID- 12577546 TI - [Reaction of regional abdominal blood flow in peritonitis to vasoactive medication based on data of selective angiography and rheography]. AB - With the help of selective arteriography of the mesenteric vessels and rheography it was found that patients with peritonitis are characterized by a reduction of blood flow in the superior mesenteric artery as a result of a spasm of the artery and its branches in the small intestine wall. This reaction is functional and can be eliminated or attenuated by purposeful measures, the most effective of them being prolonged peridural anesthesia (PPA) and droperidol injections. Thus the both methods of correction of the peritonitis dynamics (PPA and droperidol) can block up the transition of the reactive stage of peritonitis to toxic. The use of regional rheography is thought to be more expedient in medical practice than selective angiography for the assessment of the character of mesenteric blood flow in operated peritonitis patients as a more accessible and safe method for multiple performances. PMID- 12577547 TI - [New approaches to the treatment of postoperative peritonitis]. AB - A retrospective comparative analysis of results of surgical treatment of 245 patients with postoperative peritonitis was made. In 114 patients (control group) furacin was used for sanitation of the abdominal cavity, and in 131 patients (main group) sanitation of the abdominal cavity was performed with water-soluble antibacterial ointments. It was established that the proposed ointment sanitation of the abdominal cavity in combination with active surgical methods based on using programmed relaparotomy resulted in 24.1% lower lethality as compared with the traditional method of sanitation. The authors assert that under conditions of marked polyorganic insufficiency the method of programmed relaparotomy with the ointment sanitation of the abdominal cavity is not very effective while using this method before the development of systemic complications is the decisive factor of the favorable prognosis. PMID- 12577548 TI - [State of the hemostasis system in Poland's syndrome]. AB - The results of a complex investigation of the system of hemostasis in 14 women with Poland's syndrome have shown combined thrombocytic-coagulational disorders, symptoms of hypocoagulation. The functional properties were impaired against the background of the normal or moderately reduced number of thrombocytes. The data of examination, case history and results of investigation of the hemostasis system show the Poland syndrome to be a variant of mesenchymal dysplasias. Changes in a number of the indices of the thrombocytic link of hemostasis depend on the degree of the deformity. The scheme of preoperative medicamental correction has been mastered. The results of a control hemostasiological investigation have shown an improvement of hemostatic properties of the clot, normalization of the adhesive and aggregational properties of thrombocytes. Minimal hemorrhage from soft tissues was noted in operative procedures after medicamental pretreatment. PMID- 12577549 TI - [Comparative evaluation of the methods of treatment of dislocations of the clavicle]. AB - An analysis of treatment was made in 93 patients divided into three groups: conservative treatment (11 patients), operations by traditional methods (28 patients), surgery using constructions of titanium nickelide (54 operations). It was found that the conservative treatment of complete acromioclavicular end dislocation was followed by a repeated displacement, development of deforming arthrosis and deformity of the area of injury. Transcapticular fixation of acromioclavicular end dislocation resulted in the development of deforming arthrosis (68%), the incidence of this complication depending on the kind of operation. The greatest inclination of the construction to migration was noted after fixation with wires. After the application of a titanium nickelide construction the most frequent process was resorption of the bone around the construction in the shape of a hook (36%) which resulted in some patients in subluxation (11%) and development of deforming arthrosis (4.5%). PMID- 12577550 TI - [Specific features of the course of burn sepsis in children]. AB - Infectious complications after combustion represent the leading cause of lethal outcomes and attract an increasing attention of specialized services (anesthesiologists-resuscitators, combustiologists, infectionists, immunologists, clinical pharmacologists etc.). The increased incidence of the pyo-septic pathology in the whole world resulted first of all in the growing number of complications of burn disease in children. At the same time there appeared new pathological states which are thought to be outside the frame of traditional notions (e.g. the development of toxic shock syndrome). Many causes depending on the functional peculiarities of the children organism are responsible for the specific course of burn disease in children, the process itself having a number of peculiar features. PMID- 12577551 TI - [Diagnosis and characteristics of the clinical course of necrotizing enterocolitis in children]. AB - Necrotizing enterocolitis (NEC) is a disease of unknown etiology affecting mainly premature newborns and occurring in about 1 to 7% of all admissions to neonatal intensive care units. Among the greatest problems is the early diagnosis and adequate and timely treatment, surgery included. In the last quarter of the century the survival rate of premature infants was gradually increasing in national regions and worldwide and accordingly there is an increasing number of babies with NEC. To date, to treat patients with the diagnosed enterocolitis is a complicated problem. The mortality rate among the children remains high. In the period from 1978 to 2000 432 patients with NEC were admitted to the St. Petersburg Center of Neonatal Surgery in the Children Hospital N 1,300 of them requiring surgical interventions. The author's personal experience with the diagnosis and treatment of the disease allowed three main forms of the clinical course of the disease to be established: superacute, acute and subacute. PMID- 12577552 TI - [Mucosal denudation of the rectum under conditions of temporary arrest of blood flow when performing the Soave operation]. AB - The article describes the authors' positive experiences with using a simultaneous temporary arrest of blood flow in the superior rectal and iliac arteries at the stage of demucosation of the rectum in two patients when performing resection of the large intestine by the Soawe method. PMID- 12577553 TI - [Preservation of the components of blood and bone marrow for autotransfusion in patients of a particular risk contingent]. AB - Cryopreservation is the method of choice in solution of the problem of the done in good time reservation of the components of blood and bone marrow for autotransfusion in patients referred to the contingent of particular risk. Freezing of autologous hemocomponents increases the infection safety and efficiency of therapy with hemocomponents. The wide introduction of using autologous hemocomponents into the everyday surgical practice and in the arsenal of means and methods of the medicine of extreme situations is an indispensable condition for increasing the effects of curative and medico-evacuational measures. PMID- 12577554 TI - [Tracheostomy in emergency conditions in children]. AB - Clinical observations show that intubation continuing as long as 7 days are followed by complications in 37% of cases at an average. A longer intubation results in complications twice more often. Placing tracheostoma is an alternative of prolonged intubation of the trachea. In spite of the wide introduction of this method into practice, it has both the supporters and active opponents. The indications, specific features and consequences of placing tracheostoma were studied in 69 children. The assessment of the efficiency of treatment of children in critical states with the method of placing tracheostoma is given. PMID- 12577555 TI - [Use of plasma technologies in surgery of hepatic echinococcosis]. AB - During 15 years 271 patient have been operated upon in the clinic for hepatic hydatidosis. Traditional methods of operations were used in 100 of them, in 171 patients plasma technologies were applied. Plasma technologies allowed to make the number of complications 2.5-3 times rarer and the duration of postoperative treatment 18.2 bed-days shorter (51.7%). No recurrences were detected within the period from 1 month till 10 years. PMID- 12577556 TI - [Method of conducting the jejunum for hepatico-jejunal anastomosis]. AB - The authors describe a method of conducting the jejunum for hepaticojejunal anastomosis behind the colon and stomach. The authors show the advantages of the method confirmed by clinical observations and by investigations of corpses. PMID- 12577557 TI - [Surgical correction of flatfoot]. AB - The article presents results of treatment of 76 patients with transverse platypodia and the outward aberration of the first toe. In 55 of the patients (72.4%) plasty of the transverse fornix of the foot either with a vascular prosthesis or with a lavsan band was fulfilled parallel with the clinoid resection of the head of the first metatarsus. There were no recurrences of the disease in these patients during the period from 2 to 10 years, destruction of the bone in the area of a contact with the synthetic material being absolutely absent in 33 patients, who had vascular prosthesis used for the plasty. A narrow lavsan band was used in 22 patients and such destruction was revealed in 8 of them within 1-2 years. In 6 patients in whom only longitudinal marginal resection of the head of this bone with exostosis had been fulfilled a recurrence of the disease appeared in all the patients. Clinoid resection of the bone head was performed additionally in 12 patients. Recurrences were revealed in 4 of them. Good functional effect was obtained in 19 patients with a longitudinal platypodia due to the developed method of shifting the tendon of the anterior tibial muscle onto the inner part of the foot. PMID- 12577558 TI - [Lung cancer complicated by paraneoplastic state]. AB - A 10 year experience with dealing with the problem of surgical treatment of patients with lung cancer complicated by paracancrosis is described. Paracancrosis is characterized by the appearance and manifestation of different inflammatory and pyodestructive alterations in the thoracic cavity resulting from the tumor. The paracancrosis pathogenesis, its clinical symptoms, methods of diagnosis are described. The specific features of surgical treatment of patients with lung cancer complicated by paracancrosis are shown as well as the results of 5-year follow-up. PMID- 12577559 TI - [Use of drainage in endoscopic video-assisted surgical treatment of cholelithiasis]. AB - Under investigation were the nearest results of using a modified fashine drain in treatment of 57 patients operated laparoscopically for a complicated cholelithiasis. The method was shown to have both advantages and adverse effects as compared with the traditional kinds of drainage. The indications to drainage with a fashine drain after a laparoscopic procedure for complicated cholelithiasis were established and investigated. PMID- 12577560 TI - [Local treatment of purulent wounds with kukumazim]. AB - The authors describe the results of treatment of 245 patients with pyo-necrotic diseases of soft tissues with a new proteolytic enzyme of vegetable origin Kukumazim. The application of Kukumazim resulted in a 5-6 days shorter period of complete healing the wounds. No adverse effects of using Kukumazim were noted. PMID- 12577561 TI - [A rare abdominal-thoracic wound through the perineum]. PMID- 12577562 TI - [Intestinal obstruction in incarceration of internal hernia of the abdomen]. PMID- 12577563 TI - [Surgical sepsis. II. Effectiveness of immune therapy with recombinant interleukin-2]. AB - Two methods of administration of Roncoleukin were used as two subcutaneous injections in dose of 500,000 units with a 3 day interval and in the regimen of extracorporeal immunotherapy. The use of Roncoleukin resulted in the clinical improvement in 83% of patients and in the detoxicating and immunocorrecting effects in 77.3 and 60% respectively which was reliably different from analogous indices (33.4%, 45% and 7.5%) in the placebo group. Other values of the prognosed lethality being equal, the real 28-day lethality in the placebo group was 21.5%, while the using of Roncoleukin allowed the level of lethality of patients with surgical sepsis to become 3.8 times lower, including the subgroup of patients with severe sepsis from 50 to 13.6%. The trials performed showed the drug Roncoleukin to be endurable and not toxic and allowed to determine the indications and contraindications to using cytokin therapy in the complex treatment of pyo-septic diseases. PMID- 12577565 TI - [Multilayer deep hernioplasty by E. Shouldice method in inguinal hernias]. AB - The article gives a detailed description of steps of performing multilayer deep hernioplasty after E. Shouldice. In order to substantiate the possibilities to perform the procedure the structure of the transverse fascia in the ilioinguinal area was investigated in 20 corpses without hernias as compared with those having hernias. The data obtained give qualitative characteristics of the content of the collagen and elastic fibers of the transverse fascia. Their amount was proved to be 1.5-2 times less as compared with normal. Under analysis were results of surgical treatment of 786 patients with inguinal hernias. Long-term results of the surgical treatment were studied in 281 patients operated by the method of the multilayer deep hernioplasty after E. Shouldice. The method of surgical treatment of inguinal hernias was proved to be pathogenetic and highly reliable. PMID- 12577564 TI - [Immunopathogenesis of severe wounds and traumas: possibilities of immune correction]. AB - The authors describe the present-day views on the nature of immune dysfunctions in severe traumas. Based on personal clinical experiences and literature data the authors discuss the role of immune dysfunctions in pathogenesis of the traumatic disease. Special attention is given to the role of the immune system in the development of the life-threatening condition: polyorganic insufficiency whose formation mainly results from disorganization and functional failure of the system of immune reactivity. Clinical investigations have shown high effectiveness of early administration for severe wounds and traumas of a new means of immunocorrection--yeast recombinant interleukin-2 of man (preparation Roncoleukin). The administration of this immunocorrector in complex schemes of intensive therapy of the victims was shown to prevent the development of severe pyo-septic pathology and perfectly change the course of the traumatic disease. PMID- 12577566 TI - [Antibiotic prophylaxis of surgical infections]. PMID- 12577567 TI - [Oncologic markers in the diagnosis of malignant tumors of the digestive tract]. PMID- 12577568 TI - [Present-day methods of treatment of varicocele]. PMID- 12577569 TI - [Complications of open surgical procedures in urolithiasis and their prevention]. AB - 236 operative interventions for urolithiasis have been performed in 234 patients. Of them, 187 (79.9%) patients developed complications. Intraoperatively complications resulted from opening of the pleural and abdominal cavities (20 patients, 8.5%) and intraoperative blood loss < 500 ml (19 patients, 8.1%). In one case a nephrectomy was made because of repeated hemorrhage early after the operation. Among the postoperative complications most frequent were acute pyelonephritis (34, 14.5%) and aggravation of renal failure (11, 4.7%). The former demanded extracorporeal detoxication in 9 cases, in the latter hemodialysis was not required. Residual comcrements occurred in 28 (12%) patients. 17 of them had to undergo extracorporeal lithotripsy. Thus, open urological interventions in urolithiasis are now conducted in advanced disease, provoking complications. This raises responsibility of the surgeons who must take maximal preventive measures and use novel technologies in management of the complications. PMID- 12577570 TI - [Comparative characteristics of diagnostic methods of primary hyperparathyroidism in patients with urolithiasis]. AB - Nephrolithiasis (NL) is a frequent complication of hyperparathyroidism (HPT). Such patients account for 5% of those with nephrolithogenesis. The study included 89 HPT patients (age 34.8 +/- 1.9 years) with NL, 22 patients (age 38 +/- 1.4 years) with NL and 10 healthy controls (age 30 +/- 1.2 years). Hypercalciemia was seen in 27-20% of patients with HPT and NL, 9-10% of patients with nephrolithiasis. Evaluation of parathyroid hormone (PTH) levels in the blood of the examinees showed its high diagnostic value: in 89% of HPT patients this level was high. It was also elevated in 10% of NL patients. cAMP in HPT and NL was elevated in 68.48 and 12% of patients, respectively, serum concentration of Ca2+ was high in 81.4 and 24.5%, Ca load test was positive in 80.9 and 10% of the patients, respectively. It is important for diagnosis that in HPT bone mass is low. PMID- 12577571 TI - [Implications of beta 2-microglobulin in the detection of associated urinary tract diseases in patients with infiltrative pulmonary tuberculosis]. AB - The addition of radioimmunoassay of beta 2-microglobulin (beta 2-MG) from the sera of patients with infiltrative pulmonary tuberculosis to the complex of clinical, x-ray, ultrasound, cytological and biochemical examinations contributes to early detection of renal dysfunction which is important in phthisiology. Changes in urinary and blood serum values of beta 2-MG may serve as a screening test for different nephropathies. PMID- 12577572 TI - [Gastrointestinal hemorrhage in urological diseases]. AB - The authors analyse 28 cases of gastrointestinal hemorrhage (GIH) in urological diseases and after uronephrological operations, emphasize factors of uremic intoxication and relevant complications provoking DIC syndrome. Various factors leading to stress (acute blood loss, shock, sepsis) and development of immunodeficiency disturbed morphostructure of gastric and duodenal mucosa and provoked hemorrhage which was stopped most efficiently by fibrogastroduodenoscopy with coagulation of the bleeding vessel. If this operation failed, open surgery was performed. Conservative measures consisted in DIC syndrome management policy. PMID- 12577573 TI - [Reconstructive-reparative operations in injuries of the urinary tract in obstetrical, gynecologic and abdominal surgery]. AB - Different reconstructive operations were performed in 20 patients for intraoperative traumas of the urinary tract. 4 patients had injured ureter and urinary bladder. The damage was done in the course of obstetric operations (cesarean section, uterine extirpation). In 12 cases the ureter was injured in uterine extirpation for cancer (n = 4), myoma (n = 4), prolapse of the uterus, (n = 1), extirpation of uterine cervix stump (n = 1), ureteral electrocoagulation (n = 1) and adnexectomy (n = 1). In 4 cases ligation of the ureter complicated surgical interventions for cancer of the sigmoid colon (n = 1) and rectum (n = 1), diverticulosis of the colon (n = 1) and portal cirrhosis of the liver with evident cirrhosis (n = 1). Surgical policy in the treatment of intraoperative urinary tract injuries was organ-saving. Only in 3 patients with severe acute pyelonephritis surgery was two-staged with prior nephrostomy. In the rest cases primary reconstructive operations were made. Two patients with bilateral injury of the ureters after uterine extirpation have undergone transabdominal bilateral reimplantation of the ureters by Boari in Gregoir's modification. Reconstruction of pelvic ureter was often made by using a urinary bladder graft (Boari's technique). In 1 female patient with extensive vesicovaginal fistula resultant in detruzor corrugation sigmocystoplasty was made with a good result. Serious complications after the reconstruction were absent. Urinary fistulas formed in 4 cases. In 3 of them they closed without surgical intervention. In 1 patient, to close urinary fistula complicating ureterocystoanastomosis Boari's operation was conducted with a favourable outcome. Reconstructive operations saved the kidney function. PMID- 12577574 TI - [Regeneration of post-tuberculous bladder after intestinal plastic surgery]. AB - The effects of regeneration of the urinary bladder on the intestinal transplant were assessed in conducting detrusor resection after creation of an artificial reservoir for urine in 107 patients with posttuberculous microcystis. It was found that the condition of the intestinal transplant depended on severity of anatomic changes and the scope of detrusor resection. The best results of creation of a new artificial reservoir for urine are achieved if the bladder, as a cause of regenerative process and unfavourable factor for operation outcome is totally removed. PMID- 12577575 TI - [Treatment of patients with invasive bladder cancer]. AB - Radical surgery was made in 85 patients with invasive cancer of the bladder. 89.4% patients had the tumor of stage T2-T3. After the operation all 85 patients received 3 courses of adjuvant chemotherapy M-VAC in modification of the Cancer Research Center. The interval between the courses was 3 weeks. The patients were divided into two groups. The test group (n = 43) received surgical treatment, M VAC chemotherapy and immunocorrection (general magnetotherapy, phytotherapy = cyclopheron). The control group (n = 42) received only surgery and adjuvant chemotherapy. It was found that the above immunocorrection reduces the number of postoperative complications 3.8 times, side effects by chemotherapy by 1.4 times. Absolute count of T lymphocytes and their active forms elevated by 42.8 and 35.7%, respectively, while CIC diminished by 48.8%. The addition of immunocorrection to combined treatment of patients with invasive cancer of the bladder decreased the number of recurrences for 2 years twice. 3-year survival was 1.5 times longer. PMID- 12577576 TI - [Yohimbine in the treatment of erectile dysfunction]. AB - Iochimbin hydrochloride was given to 153 men with erectile dysfunction. The results are available for 140 of them. The ability of iochimbin in a single dose of 5-10 mg to enhance arterial blood inflow to cavernous bodies of the penis was confirmed by dynamic angiopenoscintigraphy and Doppler ultrasonography. Iochimbin hydrochloride in a mean daily dose of 15-20 mg proved effective in erectile dysfunction--38 to 84% responders depending on the type of erectile dysfunction. Occasional side effects can be relieved by reducing the drug dose. PMID- 12577577 TI - [Vacuum-constriction therapy in association of erectile dysfunction and premature ejaculation]. AB - Choice of treatment in combination of erectile dysfunction with premature ejaculation (PE) is not a simple task because of multifactor etiology of these diseases and physiologically antagonistic stimulating domination of parasympathetic autonomic nervous system for erectile function and sympathetic one for ejaculation. Therefore, some present-day effective treatments for each of the above diseases are contraindicated in their combination. 5 patients with the above combination gave an informed consent to vacuum-constrictor therapy (VCT) with application of ErecAid Classic Osbon unit. Control examination 6 months after VCT stated rehabilitation of the erectile function. Ejaculatory latency increased insignificantly, but erection could sustained coitus for 30 minutes. PMID- 12577578 TI - [Minimally invasive method of the treatment of prostatic abscess]. AB - In discussion of advantages and shortcomings of different surgical procedures in the treatment of a prostatic abscess (PA) which rarely complicates acute prostatitis the authors focus on a thin needle transcutaneous puncture of PA under ultrasound control. This low invasive operation made under local anesthesia was performed in 15 PA patients. The abscess cavity was drained using a polyethylene catheter "pigtail". The manipulation was well tolerated by all the patients with no complications in the postoperative period. A mean time of the abscess cavity draining was 4 days. A follow-up for 1.5 years discovered recurrence in none of the patients. This low invasive method is proposed as an alternative to the existing traumatic methods of PA management. PMID- 12577579 TI - [Complications of calculous pyonephrosis]. PMID- 12577580 TI - [Surgical treatment of bladder diverticulum and ureteral ectopy]. PMID- 12577581 TI - [Diagnostic possibilities of prostate biopsy]. PMID- 12577582 TI - [Upgrading methods of early detection of recurrent tumors of the bladder]. PMID- 12577583 TI - Neuroprotection by dietary antioxidants: new age of research. PMID- 12577584 TI - Comparison of the effects of microwave cooking and conventional cooking methods on the composition of fatty acids and fat quality indicators in herring. AB - The aim of this study was to examine the changes which occur under the influence of different heating methods in the compositions of fatty acids and selected fat quality indicators in fillets of herring. The results that are compared herein were obtained via conventional culinary techniques and using microwave radiation. Culinary processes like boiling, grilling and frying, whether done conventionally or with a microwave oven, did not lead to a reduction in the n-3 polyunsaturated fatty acids (PUFAs) fraction of the total fatty acids, indicating that these fatty acids have a high durability and a low susceptibility to thermal oxidative processes. The culinary processes used in this study also had an insignificant influence on the fat quality indicators--the peroxide and anisidine value. The fat quality indicators in herring, both after conventional and microwave heating, differ little, and indicate a low content of primary and secondary products of oxidation. PMID- 12577585 TI - Bioavailability of zinc in fiber-enriched bread fortified with zinc sulphate. AB - The present study aimed to reduce the caloric value of bread by substituting a part of wheat flour with artichoke bracts at levels of 5%, 10% and 15% without sacrificing taste, texture or acceptability. Moreover, considerable trials had been made to reduce zinc deficiency in wheat bread and fiber-enriched bread and also to study the effect of fiber on zinc bioavailability. Therefore, zinc sulphate was added to bread at levels of 40, 60, 80, 100 and 120 mg/100 g edible portion. The results from this study show that: (i) The addition of artichoke bracts to wheat flour increased the water absorption, arrival time, development time, and weakening of the dough as the level of artichoke bracts increased, while dough stability decreased. (ii) Mixing wheat flour with increasing amount of artichoke bracts increased the content of protein, fiber and total essential amino acids, also all essential amino acids increased in wheat bread and fiber enriched bread after fortification with zinc sulphate at a level of 100 mg/100 g edible portion except methionine, threonine and tyrosine. (iii) The best level of zinc sulphate to give the best bioavailability for zinc is 100 mg/100 g edible portion. (iv) Evaluation of fortified wheat bread and fiber-enriched bread with zinc sulphate showed no significant difference by test panel. PMID- 12577586 TI - Digestive stimulant action of three Indian spice mixes in experimental rats. AB - The present study examined the favourable influence of three spice mixes derived from a few commonly consumed spices of known digestive stimulant action on digestive enzymes of pancreas and small intestine, and on bile secretion and composition in experimental rats. The common ingredients of these mixes were coriander, turmeric, red chilli, black pepper and cumin, while the spice mix II additionally had ginger, and spice mix III contained onion. All the three spice mixes favourably enhanced the activities of pancreatic lipase, chymotrypsin and amylase when consumed during the diet. In addition, these spice mixes brought about a pronounced stimulation of bile flow and of bile acid secretion. Among the three spice mixes examined, spice mix III which is customized so as to include spices that are desirable from the point of view of stimulation of digestion, had the highest stimulatory influence particularly on bile secretion, bile acid output and the activities of pancreatic enzymes. While activities of pancreatic lipase, amylase and chymotrypsin were elevated by 40, 16 and 77%, respectively, the bile volume as well as the bile acid secretion were almost doubled in spice mix III treatment. The higher secretion of bile especially with an elevated level of bile acids and a beneficial stimulation of pancreatic digestive enzymes, particularly of lipase could probably be the two mechanisms by which these combinations of spices aid in digestion. PMID- 12577587 TI - Dynamic rheological properties of dough as affected by amylases from various sources. AB - The effect of alpha-amylases from cereal, fungal, and bacterial sources on dough dynamic rheological properties was investigated. Dynamic rheological study of flour-and-water doughs during resting period showed significant changes in dough rheological properties as a function of alpha-amylases. Addition of alpha amylases caused a time-dependent decrease in G', storage modulus. The enzyme action on starch during baking increased viscous flow properties. These changes were temperature-dependent. The thermal inactivation temperature of alpha-amylase plays an important role in modification of starch. Rheological changes in dough will alter the machinability of the dough and the quality of end products. PMID- 12577588 TI - Effects of incorporation of interesterified plant oils on quality and fatty acid composition of Turkish-type salami. AB - Application of interesterified plant oils (IPOs) prepared from palm, cottonseed and olive oil (IPaO, ICO and IOO, respectively) have been studied in Turkish-type salamis. Beef fat (10%) was replaced by one of the IPOs for 20, 60 or 100%. Incorporation of IPO was well reflected in fatty acid composition and saturated fatty acids/unsaturated fatty acids (SFA/UFA) ratios of the lipid fraction of all batches. Significant differences in appearance, color, texture, flavour or total score were not found among the control and Turkish-type salamis with different types and levels of IPO. We conclude that IPOs may be successfully applied as beef fat substitutes in Turkish-type salami. PMID- 12577589 TI - Effect of reuterin produced by Lactobacillus reuteri on the surface of sausages to inhibit the growth of Listeria monocytogenes and Salmonella spp. AB - Reuterin is a bacteriocin produced by some strains of Lactobacillus reuteri. The strain used in this study was isolated from raw milk from a dairy farm nearby Ankara. Beef sausage is a long years produced bratwurst style meat product in Turkey, as well as in some other countries in the Mediterranean region. Sausages are produced by raw meat; sometimes lactic starter cultures are added or spontaneous fermentation is employed. The production and storage conditions of the product promotes the growth of Listeria monocytogenes and Salmonella spp. Although nitrate is added as an antimicrobial substance against many pathogens, sometimes however nitrate application is not preventive enough on the surface because of the natural film around the sausages. Since most of the contaminations take place at post production steps, pathogenic growth is more effective on the surface of the sausages in refrigerated conditions. In this study, reuterin was applied to the surface of the sausages in order to prevent the growth of these two pathogens along with nitrate used as an additive in the product. Reuterin has inhibited the growth of L. monocytogenes considerably but not of Salmonella spp. on the surface of the sausages. PMID- 12577590 TI - Inhibition of Listeria monocytogenes LMG10470 by plantaricin UG1 in vitro and in beef meat. AB - The inhibition of Listeria monocytogenes LMG 10470 occurred due to plantaricin UG1, but not to lactic acid produced by Lactobacillus plantarum UG1 or its negatie variant (BAC). Partially purified plantaricin UG1 had a higher antilisterial activity in vitro and in meat than Lactobacillus plantarum UG1 culture. Plantaricin UG1 activity was higher in brain heart infusion (BHI) broth than in minced meat. The inhibitory effect of plantaricin UG1 against L. monocytogenes LMG10470 was dependent on its concentration. The 22,880 AU/mL appeared to be an ideal meat preservative in this experiment. PMID- 12577591 TI - Distribution of deoxynivalenol and 3-acetyldeoxynivalenol in naturally contaminated and Fusarium culmorum infected triticale samples. AB - Eight triticale genotypes, naturally contaminated (C) and collected from heads inoculated with two separate isolates Fusarium culmorum (I1 and I2) kernels were analysed for accumulation of Fusarium toxins. Triticale kernels were contaminated with deoxynivalenol (DON) and 3-acetyldeoxynivalenol (3-AcDON). Statistical analysis revealed significant differences between naturally contaminated samples (C) and those inoculated with isolate I1 and I2. The following accumulation of toxins in kernels (mean mg/kg) was found: C, I1, I2--DON 0.06, 14.45, 8.09; 3 AcDON--0.01, 0.20, 0.49. Distribution of DON and 3-AcDON was studied in four size fractions of kernels (> 2.8 mm; < or = 2.8-2.5 mm, < 2.5-2.2 mm and < 2.2 mm). The highest concentration of DON and 3-AcDON was found in the smallest fraction of damaged kernels. The percentage distribution of DON and 3-AcDON in fraction < 2.2 mm was (C) 66%, 77%, (I1) 38%, 27%, (I2) 46%, 40%; and in fraction < 2.5 mm: (C) 85%, 94%, (I1) 57%, 51%, (I2) 80%, 76%. The naturally contaminated kernel fraction < 2.2 mm (although only 2% of all) contained 33% DON and 59% 3-AcDON, for more toxigenic isolate I1 10%, 15%, 14%, less toxigenic isolate I2 4%, 13%, 11%, respectively, calculated on dry matter basis. No significant correlation between examined characteristics was found for either I1 or I2 isolate. The biosynthesis mechanism of toxic metabolites was to approximate (significantly correlated) to pair objects C/I2. PMID- 12577592 TI - Antioxidative effects of alpha-tocopherol and ascorbyl palmitate on thermal oxidation of canola oil. AB - Canola oil, with and without added alpha-tocopherol and ascorbyl palmitate, was used to deep-fat frying potatoes once per day for 10 day or once per week for 10 weeks. Changes in chemical and physical properties were monitored. Refractive index, free fatty acids and absorbance at 232 and 270 nm increased with frying time. Smoke point, 'L' color index, iodine value and the C18:2/C16:0 ratio in the oil decreased with the frying time. According to smoke point, which shows the time of the oil should be discarded, canola oil without antioxidant (control) can be used safely 8 times daily or 7 times weekly. For canola oil with antioxidant (oil + alpha-tocopherol + ascorbyl palmitate), the smoke point did not decrease below 170 degrees C in both daily and weekly fryings. This showed that canola oil with antioxidant could be used safely at least 10 times for both frying intervals. In both daily and weekly fryings, the addition of 200 ppm alpha tocopherol and 200 ppm ascorbyl palmitate increased the oxidative stability of canola oil. PMID- 12577593 TI - Cytochemical assessment of phosphopeptides derived from casein as potential ingredients for functional food. AB - Phosphopeptides derived from casein hydrolysis are suggested to have beneficial effects beyond their mere nutritive value by enhancing availability of dietary minerals, especially calcium, iron and zinc. Apart from a possible positive action the potential may exist for adverse effects that could impose restrictions to their widespread application in functional foods for human nutrition. In the present work, various case-inophosphopeptide (CPP) preparations were assessed using different human cell culture model systems in order to estimate their cytotoxic potential and their influence on epithelial properties and differentiation of human intestinal cells (Caco-2). The general cytotoxic potential of CPPs was tested using the AlphaTox NR assay. Structural and functional differentiation of intestinal Caco-2 cells was determined by measuring transepithelial electrical resistance and activity of brush border associated alkaline phosphatase over an extended cultivation period. No loss in cell viability with respect to membrane integrity could be detected, as uptake and retention of neutral red dye (AlphaTox assay) into HeLa cells was not affected by the presence of CPPs. Cytochemical assays conducted on epithelial cells (Caco-2) showed no disturbance of normal cell development and differentiation with respect to structural as well as functional differentiation markers. As all CPPs tested did not provoke any adverse effects in terms of cytotoxicity and showed no impairment of cell differentiation and monolayer integrity of human intestinal cells, it may be supposed that CPPs do not provoke a cytotoxic response in vivo in the cells assayed in the present study. PMID- 12577594 TI - Comparative study on the characterization of antisera of anti-aflatoxin B1 from rabbit and laying hen. AB - Polyclonal antisera specific for aflatoxin B1 (AFB1) were developed in rabbits and laying hens immunized with AFB1 conjugated to bovine serum albumin (AFB1 BSA). During the immunization schedule, AFB1-specific antisera from rabbits started to obviously be secreted after 60 days of the first AFB1-BSA injection, reached to a peak at day 120, and started to go down at day 135; while in regard to laying hens, antibodies began to largely be produced at day 90, arrived to a peak at day 135, and began to go down at day 165. The rabbits consistently produced anti-AFB1 antisera in 2-fold higher amounts than the laying hens during the antisera production process. But rabbits' and laying hens' antisera to AFB1 were found not to be apparently different in cross-reaction with aflatoxin analogs. Both rabbits' and laying hens' antisera could be used as immunological reagents for detecting AFB1 in agricultural commodities. PMID- 12577595 TI - Under the radar. PMID- 12577596 TI - Choice and the post-Roe generation. PMID- 12577597 TI - How to sell XXXL. PMID- 12577598 TI - How Bush gets his way on the environment. PMID- 12577600 TI - Bedtime for baby. PMID- 12577599 TI - Sharkless seas. PMID- 12577601 TI - Where's the proof? PMID- 12577603 TI - Devil in the details? PMID- 12577602 TI - The cost of a cure. PMID- 12577604 TI - [Tachyarrhythmias--when rapid pace is pathological]. AB - Tachycardias are classified as supraventricular and ventricular tachycardias. Supraventricular and ventricular tachycardias may occur as a complication of almost any underlying cardiac disease and many extracardiac causes. In addition, patients without any detectable structural heart disease may present with these arrhythmias. In this overview, we discuss the clinical presentations, the pathophysiological mechanisms and the therapeutical strategies in patients with tachyarrhythmias. PMID- 12577605 TI - [Hyperthyroidism--differential diagnosis and differential therapy]. AB - Thyrotoxicosis is one of the most common endocrine disorders, affecting approximately 2% of the adult population. Graves' disease and its variants are the most frequent cause. The prevalence of toxic nodular goitre and toxic adenoma is increased in areas of limited iodine supply. Thyrotoxicosis in patients with multinodular goitre can be precipitated by iodinated radiocontrast agents and the antiarrhythmic drug amiodarone. Transient hyperthyroidism may be caused by HCG induced thyroiditis in pregnancy or de Quervain subacute thyroiditis. The differential diagnosis and the treatment principles of the varied causes of thyrotoxicosis are reviewed. PMID- 12577606 TI - [Venous stasis--chronic leg ulcer]. AB - Chronic venous leg ulcer is a major health problem, affecting principally the elderly. The natural history of the disease is of a continuous cycle of healing and breakdown over decades and recurrence is common. In the present manuscript a survey of the management of venous leg ulcers is given. PMID- 12577607 TI - [Tachypnea--when breathing induces panic]. PMID- 12577608 TI - Structure-property correlation and estimation of the antiparasitic activity of glutathione derivatives. PMID- 12577609 TI - On the effect of diazoxide on isolated mitochondria from rat myocardium. PMID- 12577610 TI - Changes in the ratio between the peaks of red chlorophyll fluorescence in leaves of Populus balsamifera during vegetation. PMID- 12577611 TI - Specific effects of gamma-radiation on the fine structure of the photosynthetic apparatus: evaluation of the character of disturbances in vivo using high-order derivative spectrophotometry. PMID- 12577612 TI - The role of chromatin structural changes in acquired radioresistance of cells. PMID- 12577613 TI - The study of the conditions of in vitro accumulation of a thermostable comuton from rat liver. PMID- 12577614 TI - Temperature dependence of HpRad51, the central protein of the homological recombination in the yeast Hansenula polymorpha. PMID- 12577615 TI - Kinetics of photoinhibition and delayed fluorescence in the plant photosynthetic system. PMID- 12577617 TI - Biophysical aspects of the biosphere impact on global climate. PMID- 12577616 TI - Interaction between the platelet IIb/IIIa receptor gene and serotonin transporter gene is involved in the formation of the predisposition to myocardial infarction in young men. PMID- 12577618 TI - The GABAergic midbrain system is involved in the control of sleep and temperature homeostasis in pigeons. PMID- 12577619 TI - Artificial mycobacterial particles for immunization against tuberculosis. PMID- 12577620 TI - Effects of brain peptides on instrumental learning in monkeys Macaca fascicularis. PMID- 12577621 TI - The location capacity of dolphins in evaluation of speed characteristics of targets and technical apparatuses for hydrolocation and radiolocation. PMID- 12577622 TI - The mitochondrial antioxidants thioredoxin-2 and Mn-superoxide dismutase are involved in the mechanisms of brain hypoxic tolerance. PMID- 12577623 TI - Effect of cold adaptation of rabbits on the alpha 1- and beta-adrenergic blood pressure responses of intestinal blood vessels. PMID- 12577624 TI - Specific anosmia to isovaleric acid in laboratory C57BL/6 mice: recessive inheritance. PMID- 12577625 TI - The period of the sensitivity to the pheromone inhibiting spermatogenesis in laboratory CBAB6F1 mice. PMID- 12577626 TI - From studies of the location capacity of dolphins to development of technical devices for hydrolocation and radiolocation. PMID- 12577627 TI - Coding of efferent signals in monkey neostriatum. PMID- 12577629 TI - The effect of the wolf on the competition between ungulates in the Voronezh Biosphere Reserve. PMID- 12577628 TI - Variable social rank of white-spotted male water voles: the effect of the presence of females. PMID- 12577630 TI - Temperature conditions for development of two species of malaria pathogens in the vector organism in Russia in the 20th century. PMID- 12577631 TI - Interaction of wheat lectin with 24-epibrassinolide in the regulation of cell division in wheat roots. PMID- 12577632 TI - The use of RAPD analysis of sugar-beet mitochondrial DNA in search of CMS genes. PMID- 12577633 TI - Two types of rDNA internal transcribed spacers (ITSs) in the Capsicum genome. PMID- 12577634 TI - Interspecies diversity of the genome responses to chronic irradiation in natural populations of rodents. PMID- 12577635 TI - Relationships between the individual growth rate and the population spatial structure and dynamics in rodents. PMID- 12577636 TI - Saltatory changes in the structure of the ribosomal DNA external transcribed spacer during the evolution of cockroaches of genus Blattella. PMID- 12577637 TI - RAMD and the phylogeny of pines of the Strobus section. PMID- 12577638 TI - M-protein as a possible inducer of depressive state in influenza. PMID- 12577639 TI - Characteristics of development and ossification of skeletal elements in the limbs of the smooth newt Triturus vulgaris (Caudata). PMID- 12577640 TI - On the role of some checkpoint genes in determination of the radiation sensitivity of the yeast Saccharomyces cerevisiae. PMID- 12577641 TI - Specific structure of root cells of the salt-accumulating halophyte Suaeda altissima L. PMID- 12577642 TI - Identification of the pea (Pisum sativum L.) genome chromosomes using C-banding analysis. PMID- 12577643 TI - Salicylic acid induces dissipation of the proton gradient on the plant cell plasma membrane. PMID- 12577644 TI - A hybridization study of different geographical populations of the predatory yeast Arthroascus schoenii. PMID- 12577645 TI - Specific features of the development of isolated meristems of Arabidopsis thaliana (L.) Heynh. PMID- 12577646 TI - Immunocytochemical reorganization of the nucleolus in human embryo fibroblasts infected with cytomegalovirus in vitro. PMID- 12577647 TI - Detection of a new avian hepadnavirus with an unexpectedly broad host range. PMID- 12577648 TI - The dining experience. Making it pleasurable for long-term care residents. PMID- 12577649 TI - Education of residents (PF). PMID- 12577650 TI - January themes. PMID- 12577651 TI - I was greedy, too. AB - Americans are outraged at the greediness of Wall Street analysts, dot-com entrepreneurs, and, most of all, chief executive officers. How could Tyco's Dennis Kozlowski use company funds to throw his wife a million-dollar birthday bash on an Italian island? How could Enron's Ken Lay sell thousands of shares of his company's once high-flying stock just before it crashed, leaving employees with nothing? Even America's most popular domestic guru, Martha Stewart, is suspected of having her hand in the cookie jar. To some extent, our outrage may be justified, writes HBR senior editor Diane Coutu. And yet, it's easy to forget that just a couple years ago these same people were lauded as heroes. Many Americans wanted nothing more, in fact, than to emulate them, to share in their fortunes. Indeed, we spent an enormous amount of time talking and thinking about double-digit returns, IPOs, day trading, and stock options. It could easily be argued that it was public indulgence in corporate money lust that largely created the mess we're now in. It's time to take a hard look at greed, both in its general form and in its peculiarly American incarnation, says Coutu. If Federal Reserve Board chairman Alan Greenspan was correct in telling Congress that "infectious greed" contaminated U.S. business, then we need to try to understand its causes--and how the average American may have contributed to it. Why did so many of us fall prey to greed? With a deep, almost reflexive trust in the free market, are Americans somehow greedier than other peoples? And as we look at the wreckage from the 1990s, can we be sure it won't happen again? PMID- 12577652 TI - Why bad projects are so hard to kill. AB - Even at the prototype stage, experts were saying the technology was obsolete. Yet, in the face of tepid consumer response, the company stubbornly kept increasing production capacity and developing new models. By the time it was finally killed, the initiative had cost the company an astounding $580 million and had tied up resources for 14 years. The product was RCA's SelectaVision videodisc recorder, and its story is hardly unique. Companies make similar mistakes--if on a somewhat smaller scale--all the time. But why? No one comes to work saying, "I'm going to pursue a project that will waste my company millions of dollars." Quite the opposite. They come to work full of excitement about a project they believe in. And that, surprisingly, can be the root of all the trouble--a fervent belief in the inevitability of a project's ultimate success. Starting, naturally enough, with a project's champion, this faith can spread throughout the organization, leading everyone to believe collectively in the product's viability and to view any signs of impending doom merely as temporary setbacks. This phenomenon is documented here in two chillingly detailed case studied, one involving Essilor, the world's largest maker of corrective lenses for eyeglasses, and the other involving Lafarge, the largest producer of building materials. By counterexample, they point the way toward avoiding such morasses: assembling project teams not entirely composed of like-minded people and putting in place--and sticking to--well-defined review processes. Both cases also show that if it takes a project champion to get a project up and running, it may take a new kind of organizational player--an "exit champion"--to push an irrationally exuberant organization to admit when enough is enough. PMID- 12577653 TI - Who's bringing you hot ideas and how are you responding? AB - There's an unsung hero in your organization. It's the person who's bringing in new ideas from the outside about how to manage better. These aren't your product and service innovators--those people are celebrated loudly and often. This is the manager who, for instance, first uttered the phrase "balance scorecard" in your hallways, or "real options," or "intellectual capital." Managerial innovation is an increasingly important source of competitive advantage--especially given the speed with which product innovations are copied--but it doesn't happen automatically. It takes a certain kind of person to welcome new management ideas and usher them into an organization. The authors recently studied 100 such people to find out how they translate new ideas into action in their organizations. They discovered that they are a distinct type of practitioner; that is to say, they resemble their counterparts in other organizations more than they resemble their own colleagues, and they share a common way of working. "Idea practitioners," as the authors call them, begin by scouting for ideas. All of them are avid readers of management literature and enthusiastic participants in business conferences; many are friendly with business gurus. Once they've identified an idea that seems to hold promise, they tailor it to fit their organizations' specific needs. Next, they actively sell the idea--to senior executives, to the rank and file, to middle managers. And finally, they get the ball rolling by participating in small scale experiments. But when those take off, they get out of the way and let others execute. In this article, the authors identify the characteristics of idea practitioners and offer strategies for managing them wisely. PMID- 12577654 TI - Negotiating the spirit of the deal. AB - Most experienced negotiators are comfortable working out the terms of an economic contract--they bargain for the best price, haggle over equity splits, and finesse detailed exit clauses. Yet these same seasoned professionals spend so much time ironing out the letter of the deal that they often pay little attention to the spirit of the deal--the social contract. And that can lead to major problems, say the authors, because even though the parties agree to the same terms on paper, they may have very different expectations about how to meet them. Those on one side, for instance, might think they're entering into a long-term partnership, while those on the other believe they're simply making a series of discrete transactions. Because the parties have failed to have a true meeting of the minds, they sign a deal that is likely to fall apart. To avoid such a disastrous outcome, negotiators should explicitly discuss the details of their social contract before inking the deal. They should talk about the underlying social contract, which answers the question, What? For instance, what is the real nature, extent, and duration of the agreement? And they should discuss the ongoing social contract, which answers the question, How? For instance, in practice, how will we make decisions, handle unforeseen events, communicate with one another, and resolve disputes? Drawing on real-life examples, the authors explore the problems that arise when the letter and spirit of the deal are at odds and suggest ways to dovetail them so they are both independently strong and mutually reinforcing. They also highlight risk factors that can lead to misunderstandings and expose common misperceptions about the social contract. PMID- 12577655 TI - AIDS is your business. AB - If your company operates in a developing country, AIDS is your business. While Africa has received the most attention, AIDS is also spreading swiftly in other parts of the world. Russia and Ukraine had the fastest-growing epidemics last year, and many experts believe China and India will suffer the next tidal wave of infection. Why should executives be concerned about AIDS? Because it is destroying the twin rationales of globalization strategy-cheap labor and fast growing markets--in countries where people are heavily affected by the epidemic. Fortunately, investments in programs that prevent infection and provide treatment for employees who have HIV/AIDS are profitable for many businesses--that is, they lead to savings that outweigh the programs' costs. Due to the long latency period between HIV infection and the onset of AIDS symptoms, a company is not likely to see any of the costs of HIV/AIDS until five to ten years after an employee is infected. But executives can calculate the present value of epidemic-related costs by using the discount rate to weigh each cost according to its expected timing. That allows companies to think about expenses on HIV/AIDS prevention and treatment programs as investments rather than merely as costs. The authors found that the annual cost of AIDS to six corporations in South Africa and Botswana ranged from 0.4% to 5.9% of the wage bill. All six companies would have earned positive returns on their investments if they had provided employees with free treatment for HIV/AIDS in the form of highly active antiretroviral therapy (HAART), according to the mathematical model the authors used. The annual reduction in the AIDS "tax" would have been as much as 40.4%. The authors' conclusion? Fighting AIDS not only helps those infected; it also makes good business sense. PMID- 12577656 TI - The enemies of trust. AB - Researchers have established that trust is critical to organizational effectiveness. Being trustworthy yourself, however, does not guarantee that you are capable of building trust in an organization. That takes old-fashioned managerial virtues like consistency, clear communication, and a willingness to tackle awkward questions. It also requires a good defense: You must protect trust from its enemies. Any act of bad management erodes trust, so the list of potential enemies is endless. Among the most common enemies of trust, though, are inconsistent messages from top management, inconsistent standards, a willingness to tolerate incompetence or bad behavior, dishonest feedback, a failure to trust others to do good work, a tendency to ignore painful or politically charged situations, consistent corporate underperformance, and rumors. Fending off these enemies must be at the top of every chief executive's agenda. But even with constant vigilance, an organization and its leaders will sometimes lose people's trust. During a crisis, managers should enlist the help of an objective third party--chances are you won't be thinking clearly--and be available physically and emotionally. If you "go dark" in the face of a crisis, employees will worry about the company's survival, about their own capacity to cope, and about your abilities as a leader. And if trust has broken down so badly that your only choice is to start over, you can do so by figuring out exactly how the breach of trust happened, ascertaining the depth and breadth of the loss, owning up to the loss instead of downplaying it, and identifying as precisely as possible the specific changes you must make to rebuild trust. PMID- 12577657 TI - Clueing in customers. AB - Leonard L. Berry and Neeli Bendapudi When customers lack the expertise to judge a company's offerings, they naturally turn detective, scrutinizing people, facilities, and processes for evidence of quality. The Mayo Clinic understands this and carefully manages that evidence to convey a simple, consistent message: The needs of the patient come first. From the way it hires and trains employees to the way it designs its facilities and approaches its care, the Mayo Clinic provides patients and their families concrete evidence of its strengths and values, an approach that has allowed it to build what is arguably the most powerful brand in health care. Marketing professors Leonard Berry and Neeli Bendapudi conducted a five-month study of evidence management at the Mayo Clinic. They interviewed more than 1,000 patients and employees, observed hundreds of doctor visits, traveled in the Mayo helicopter, and stayed in the organization's many hospitals. Their experiences led them to identify best practices applicable to just about any company, in particular those that sell intangible or technically complex products. Essentially, the authors say, companies need to determine what story they want to tell, then ensure that their employees and facilities consistently show customers evidence of that story. At Mayo, the evidence falls into three categories: people, collaboration, and tangibles. The clinic systematically hires people who espouse its values, and its incentive and reward systems promote collaborative care focused on the patient's needs. The physical environment is explicitly designed for its intended effect on the patient experience. In almost every interaction, an organization's message comes through. "Patients first," the Mayo Clinic's message, is not the only story a medical organization could tell, but the way in which Mayo manages evidence to communicate this message is an example to be followed. PMID- 12577658 TI - Who needs budgets? AB - Budgeting, as most corporations practice it, should be abolished. That may sound radical, but doing so would further companies' long-running efforts to transform themselves into developed networks that can nimbly adjust to market conditions. Most other building blocks are in place, but companies continue to restrict themselves by relying on inflexible budget processes and the command-and-control culture that budgeting entails. A number of companies have rejected the foregone conclusions embedded in budgets, and they've given up the self-interested wrangling over what the data indicate. In the absence of budgets, alternative goals and measures--some financial, such as cost-to-income ratios, and some nonfinancial, such as time to market-move to the foreground. Companies that have rejected budgets require employees to measure themselves against the performance of competitors and against internal peer groups. Because employees don't know whether they've succeeded until they can look back on the results of a given period, they must use every ounce of energy to ensure that they beat the competition. A key feature of many companies that have rejected budgets is the use of rolling forecasts, which are created every few months and typically cover five to eight quarters. Because the forecasts are regularly revised, they allow companies to continuously adapt to market conditions. The forecasting practices of two such companies, both based in Sweden, are examined in detail: the bank Svenska Handelsbanken and the wholesaler Ahlsell. Though the first companies to reject budgets were located in Northern Europe, organizations that have gone beyond budgeting can be found in a range of countries and industries. Their practices allow them to unleash the power of today's management tools and realize the potential of a fully decentralized organization. PMID- 12577659 TI - [Comparative assessment of the diagnostic capacities of PET with 18FDG and 1 [11C] sodium butyrate in the examination of patients with tumors of the brain and cerebral circulatory disorders: results of semiquantitative assessment]. AB - The purpose of the present study was to determine the capacities of differential diagnosis of the bulky masses of the brain by positron emission tomography (PET) with 11C-butyrate by the direct indication--the tumor RFP level measured by means of the semiquantitative indicator--the differential accumulation coefficient (DAC). For comparison, 18FDG PET widely used to detect malignant tumors was preliminarily made. Brain PET was performed in 86 patients (45 males and 41 females whose age ranged from 18 to 69 years to identify bulky masses or to rule out continuous tumor growth. In all cases, the data were histologically and morphologically verified. Of the 86 patients, 21 were found to have malignant tumors, 41 had benign tumors, arteriovenous malformations, cerebral circulatory disorders, and cysts were detected in 3, 7, and 14 cases, respectively. The level of 18FDG accumulation was ascertained to be directly related to the grade of malignancy. In 20 of the 21 patients with malignant tumors, DAC for 18FDG was greater than 1. However, it was impossible to differentiate benign tumors and non tumoral masses by using only 18FDG. Comparing the data obtained by means of 18FDG and 11C-butyrate revealed their comparability in detecting neoplasms. Patients with vascular tumors (benign meningioma and adenoma of the pituitary were an exception. Their DAC for 11C-butyrate was greater than 1. In follow-up CT scanning, just a single injection of 11C-butyrate may allow one to estimate the vascular and tissue components of masses, which facilitates the identification of vascular non-tumoral processes and tumors. The additional criterion that allows a neoplasm to be differentiated from nontumoral processes permits a rapid tumor release of a radioactive label. PMID- 12577660 TI - [Comparative assessment of resting myocardial perfusion in patients with postinfarct cardiosclerosis by electron-beam tomography and 99mTc-MIBI myocardial single-photon tomography]. AB - The purpose of the study was to compare myocardial perfusion assessed by electron beam computed tomography (EBCT) with that obtained by 99mTc-sestamibi single photon emission computed tomography (SPECT) in patients with old myocardial infarction and control subjects at rest. A total of 42 patients with suspected and known ischaemic heart disease (IHD) were included in the study. 20 pts had a history of Q-wave myocardial infarction (MI), 12 pts had an old non-Q-wave MI and 10 served as controls (without perfusion defects on SPECT images at rest). Assessment of the myocardial perfusion by EBCT was performed using the short axis view and multislice mode (MSM) during injection of 50 ml of the nonionic contrast medium at 4 ml/s via cubital vein. Perfusion defects were localized by SPECT according to 6-segment model of the LV (septal, anterior, lateral, posterior, inferior and apical). Overall concordance between EBCT and SPECT was 67% for normal versus abnormal perfusion. Agreement between the 2 methods for each of the 6 segments was 81% (K = 0.62) for the anterior segment, 71% (K = 0.42) for the septal segment, 71% (K = 0.43) for the apical, 69% (K = 0.3) for the lateral segment, 48% (K = 0.13) for the posterior segment and 60% (K = -0.13) for the inferior segment. Discrepancies between the two of techniques were most notable in the posterior region. Beam hardening during passage of the contrast medium through the heart chambers and descending aorta is possible explanation of the artifacts on EBCT images. This study demonstrates that nowadays EBCT is not yet alternative to SPECT in the assessment of the myocardial perfusion in patients and further improvements of scanning techniques are necessary. PMID- 12577661 TI - [Radiation diagnosis of uncomplicated compression spinal fractures in children]. AB - Experiments on rabbits compared the X-ray, morphological, and magnetic resonance imaging of compression fractures of the vertebral body. Edema of the bone marrow of the vertebral body was ascertained to be a basic morphological substrate that evokes a modified MR signal. The data on 178 children with suspected compression fracture of the vertebral body were used to consider the potentialities of MRI and X-ray study in this pathology. The MRI semiotics of uncomplicated compression fractures of vertebral bodies is presented. The sensitivities of MRI and spondylography in the diagnosis of compression fractures of vertebral bodies were 100 and 62.5%, respectively. PMID- 12577662 TI - [Comparative assessment of percutaneous transhepatic cholangiodrainage under roentgenoscopy and ultrasound guidance]. AB - In 1990-2000, percutaneous transhepatic cholangiodrainage was performed in 89 patients with obstructive jaundice of tumoral etiology: Groups A (n = 21) under roentgenoscopic guidance and Group B (n = 68) under ultrasound guidance (USG). The efficiency and safety of puncture of the biliary tract were comparatively evaluated in these groups. The technical success of the procedure was achieved in 81.0% of Group A patients and in 98.5% of Group B ones. USG decreased radiation load on the patient and medical staff by three times and the number of needle passes by two times. External-and-internal cholangiodrainages under USG and roentgenoscopy were performed in 55.2 and 23.8%, respectively. The incidence of complications and mortality were 5.9 and 1.5% in Group B and versus 17.6% and 4.8% in Group A. It is concluded that transhepatic cholangiodrainage under USG has some advantage over puncture of the biliary tract under roentgenoscopy. PMID- 12577663 TI - [Complex use of radiation diagnostic techniques in tumors of the pancreatoduodenal area]. AB - In the work are summarized the results of the examination of the 500 patients with tumors of pancreas (141), tumors of biliary ducts (9), tumor of duodenum (1), chronic pancreatitis (315), choledocholithiasis (12) and non-specific biliary dilatation (13). The are show the possibilities different methods of radiation diagnosis (ultrasonography, CT, MRI, MR cholangiopancreatography) in the differential diagnosis tumors of pancreatico-duodenal region, estimation of the localization, specification, degree of invasion into the neoplasms gathering round the cells. The paper presents algorithm, used in the Regional hospital of Chelyabinsk. PMID- 12577664 TI - [Capacities of transabdominal ultrasound study in the diagnosis of polyps of the stomach and colon]. AB - The data obtained from studies of 84 patients with polyps of the stomach and colonic were used to consider the capacities of transabdominal and transrectal ultrasound studies and their diagnostic efficiency in the diagnosis of polyps of the stomach and colon. The ultrasound semiotics of polyps of the gastric and colon was described. A place of transabdominal ultrasound sonography (US) in the algorithm of radiation and endoscopic studies was established. This investigation led to the conclusion that transabdominal US, along with indirect diagnosis, permits solution of number of fundamental problems facing prior to polypectomy- these are primarily to define the nature of a detected polyp (epithelial and non epithelial), based on an analysis of the laminar structure of the wall of an organ at the polyp base, a possible polyp malignancy and the degree of invasion into the depths of the gastric wall, as well as to detect a feeding vessel (in the CDC mode) and to predict the likelihood of possible complications. It is advisable to use transabdominal US in a complex of diagnostic techniques for polyps of the stomach and colon. PMID- 12577665 TI - [Computed tomographic variants of vertebrogenic lumbosacral radicular lesions in patients with radicular and non-radicular lumbar ischialgias]. AB - The paper considers the CT features of vertebrogenic lumbosacral radicular lesions in patients with radicular and nonradicular lumbar ischialgias. Ninety patients were divided into three groups according to the degree of clinical manifestations of radicular disorders. Four CT patterns of radicular alterations were identified: dislocation, compression, aggregation, and conglomeration. There were differences in the frequency of different CT patterns in patients from different clinical groups. Combining the CT patterns of radicular lesions into occupational and contact types determined a correlation of the severity of radicular disorders and the occupational type of changes in the root. PMID- 12577666 TI - [Medical and economic aspects of digital roentgenography]. PMID- 12577667 TI - [Current potentialities of radiation diagnosis of temporal lobe epilepsy]. PMID- 12577668 TI - [Stenting in gastroenterology]. PMID- 12577669 TI - [A mathematical model for prediction of the effect of hyperlipidemia on thickness of the intima-media complex in the carotid artery wall as an indicator of atherosclerosis]. AB - The growth of intima media complex is widely accepted parameter in studies that are in relation to the process of atherosclerosis. Increased value of intima media thickness (IMT) a. carotis communis is the incontestable index of atherosclerosis process in regard to distinct vascular spheres and wider cardiovascular risk. Hypercholesterolemia as the well-known promoter of atherogenesis influence on the increase of IMT. It is established that there is a highly valuable connection (p < 0.001) between the levels of cholesterol and the thickness of IMT. It is also observed a highly significant linear and cubic correlation between the cholesterol and IMT. Concerning the relationship between trigliceride and IMT, it stands out as the highly relevant (R = 0.42; p < 0.001) but the coefficient of correlation is fewer than cholesterol (R = 0.56). Increased value of cholesterol and triglicerides exerts influence on IMT of a. carotis communis simultaneously which leads to highly consequential equation: IMT = 0.005 x TG-0.008 x Chol + 0.28. This mathematics model can be noteworthy if one anticipates effects of primary and secondary prevention. PMID- 12577670 TI - [Evaluation of representative blood pressure measurement in children on hemodialysis]. AB - The diagnosis of hypertension in patients on chronic haemodialysis is not easy because the blood pressure changes as a consequence of therapy (haemodialysis) and attenuated circadian rhythm of blood pressure is often present. The estimation of representative blood pressure levels is difficult because it is not known whether blood pressures measured before starting or after completion of haemodialysis are predictive for the average interdialytic blood pressure. Our group of patients consisted of 18 children with endstage renal failure, aged 13.5 +/- 3.2 years, treated with chronic haemodialysis from 0.1 to 110 (mean 25.8, median 19) months. Ambulatory blood pressure monitoring was performed during 44 h interdialytic period. Blood pressure was also measured with sphygmomanometer before starting and after completion of heamodialysis. The average blood pressure values for the last ten haemodialyses were evaluated as well. Multiregression analysis showed significant correlation between interdialytic blood pressure (systolic and diastolic) and blood pressures measured before haemodialysis (r = 0.74; p < 0.001, r = 0.78; p < 0.001, respectively) and after haemodialysis (r = 0.76; p < 0.01, r = 0.6; p < 0.05, respectively). Statistically high correlation between these blood pressures was also confirmed for the average blood pressure values for the last 10 haemodialyses (r = 0.78; p < 0.001, r = 0.75; p < 0.001 and r = 0.78; p < 0.001, r = 0.86; p < 0.001, respectively). Our findings show that blood pressures measured before starting or after completion of haemodialysis give good information about interdialytic blood pressure in children on chronic haemodialysis. PMID- 12577671 TI - [Extracorporeal shock wave lithotripsy in the urinary tract in patients with one kidney]. AB - Extracorporeal Shock Waves Lithotripsy (ESWL) is a method of choice in the treatment of lithiasis. Surgical treatment is reduced to minimum, which in our case means 1-2%. Contraindications for using this method are: blood coagulation disorders, manifest tuberculosis, pregnancy, overobesity. Thanks to appropriate indications and technique which is used in an adequate way as well as to various endurological manipulations, removal of a stone by this method is safe, trauma is minimised, which is very important for patients with one kidney. During the last three years, usually in hospital conditions, 57 patients were treated in this way. Removal of a stone was performed by Extracorporeal Shock Waves Lithotripsy as mono therapy in 30 (53%) patients; Extracorporeal Shock Waves Lithotripsy by using Double-J catheter in 16 (28%) patients; Extracorporeal Lithotripsy with urine derivation by percutaneous nephrostome in 11 (19%) patients. Endurological methods were used in 27 patients. Disintegration of stone was performed in the proper way. Full success was reached. Two patients had incrustation of ureteral catheter which had to be removed by surgery. In two other patients with inferior function of one kidney, after brief obstruction, haemodialisis was performed. Our results confirm that this method is nonaggressive, technically perfect for disintegration of urineorgans stone on all levels, followed by small complications which are often solved by endoscopic manipulations. PMID- 12577672 TI - [Multivascular disease in clinical practice]. AB - Multiple arterial disease is presented by coexistence of ischaemic heart disease, carotid disease and peripheral obliterate arterial disease. Atherosclerosis is the main factor for onset of the disease. Among 150 patients with clinical manifestations of obliterate disease of at least two aforementioned arterial systems, we examined by many noninvasive and invasive procedures the existence and degree of obliterate arterial disease of coronary, carotid and peripheral arteries of the lower extremities. The results revealed the statistically significant correlation among: ischaemic heart disease and carotid disease (r = 0.939; p < 0.01); ischaemic heart disease and peripheral arterial disease (r = 0.834; p < 0.05); ischaemic heart disease, peripheral arterial disease and carotid disease (r = 0.986; p < 0.01). The results pointed out that whenever clinical manifestations of obliterate disease of peripheral arteries are present, there is also need for routine examination of existent coronary artery disease. PMID- 12577673 TI - [Abdominal pregnancy with the placenta located on the spleen left in situ causing subphrenic abscess]. AB - Abdominal pregnancy appears once in 3000 pregnancies. It usually terminates with abortion and urgent surgery. Thanks to ultrasonography and computed tomography the diagnosis is possible before surgery. Most frequently the diagnosis has been established during emergency laparotomy. Gynaecologists are not in agreement whether removal of placenta is mandatory or not, as it may include removal of parts or entire organs or may be followed with serious bleeding difficult to control. We present a 21-year old woman in whom during an urgent laparotomy performed for abdominal pregnancy placenta inserted in the spleen was left in situ. Postoperatively the patient developed subphrenic abscess which could not be solved without reoperation during which both the placenta and the spleen were removed. Ten years after surgery she is symptom-free. PMID- 12577674 TI - [Corticoid-sensitive nephrotic syndrome in children with myelodysplastic syndromes]. AB - Several reports have documented various forms of glomerular diseases in adults with myelodysplastic syndromes (MDS), but similar reports in children are lacking. We describe two children with MDS-associated with steroid-responsive nephrotic syndrome (NS). Patient 1, who had MDS with myelofibrosis, presented also hepatosplenomegaly, pancytopenia, chronic hepatitis, moderate proteinuria, hypocomplementamia and elevated ANA titer. During initial prednisone treatment proteinuria markedly diminished and partial but transient haematological improvement occurred. Relapse subsequently occurred that was manifested by overt NS and pancytopenia. High doses of prednisolone led to remission of the renal disease but haematological remission did not occur. Persisting pancytopenia and repeated infections terminated in sepsis, two years after the onset of MDS. Patient 2, who had refractory anaemia with clonal monosomy 19, manifested bowel disease, hepatosplenomegaly, anaemia and non-organic specific autoantibodies. Prednisone led to both clinical and haematological remission. Haematologic disease relapsed 12 months later, when nephrotic-range proteinuria, haematuria and mild azotaemia were also found. Corticosteroid treatment led to long-lasting renal and haematologic remission, maintained by a small dosage of prednisone. In both patients, renal biopsy findings were consistent with those seen in idiopathic NS. A Medline search disclosed 16 cases of glomerulopathy in the course of MDS in adult patients. Clinical features included NS, usually accompanied by renal insufficiency with either acute, chronic, or rapidly progressive glomerulonephritis. On biopsy, membranous nephropathy, crescentic or mesangial proliferative glomerulonephritis and AL amyloidosis, were found. We conclude: (1) that glomerular disease may be present and should be searched for in patients with MDS; (2) that MDS can be added to the list of rare conditions associated with corticosteroid-responsive NS in children. PMID- 12577675 TI - [Duodenal obstruction due to chronic pancreatitis]. AB - Chronic pancreatitis may cause a number of obstructive complications. Obstructions of the pancreatic and common bile duct are the most frequent, obstruction of the duodenum is rare and obstruction of the colon is very rare. Duodenal stenosis caused by chronic pancreatitis is usually moderate so that the surgical treatment is not necessary except if operation is necessary for other obstructive complications. The severe isolated duodenal stenosis for which surgery is mandatory is rare. We present a 46-year old male suffering from chronic alcoholic pancreatitis causing almost complete duodenal obstruction in whom surgical treatment was necessary. The truncal vagotomy and gastroenterostomy were performed. With abstinence of alcohol, low fat diet and substitute therapy more than 3 years after surgery the patient is symptom-free. He regained the previous weight and shows tendency to become fat. PMID- 12577676 TI - [Allergic bronchopulmonary aspergillosis]. PMID- 12577677 TI - [Asymptomatic carotid disease--early detection]. PMID- 12577678 TI - [Magnesium supplementation during pregnancy and lactation]. PMID- 12577679 TI - [Role of postoperative radiotherapy in the treatment of early breast carcinoma]. PMID- 12577680 TI - [Dr. Svetislav Sp. Barjaktarovic--30 years later]. AB - Doctor Svetislav Sp. Barjaktarovitsh (1893-1971), full professor of University School of Medicine in Belgrade, Head of the Department of Gynaecology and Obstetrics since its foundation, Director of the University Teaching Hospital of Gynaecology and Obstetrics from 1937 to 1948, was dismissed from University for political reasons in December 1948. He was the author of classical gynaecological and obstetrics textbooks, popular books, as well as of fifty scientific papers published in domestic and foreign professional literature. While being the Director of the University Teaching Hospital of Gynaecology and Obstetrics he managed to raise money to start the building of a new hospital. He was engaged in experimental work and improved the diagnosis and therapy of genital tuberculosis, extrauterine pregnancy, adnexal inflammation, puerperal fever, etc. He was especially concerned with the determination of the child's sex before his/her birth. As a medical student he participated in the First World War working in the Company Hospital of the Morava Division. He was holder of the Albanian Commemorative Medal, The Cross of Mercy and other decorations. PMID- 12577681 TI - [Lifetime achievements of Ljubodrag T. Mihailovic. 1926-1974]. AB - Ljubodrag-Buba Mihailovitsh was born on 27th February 1926 in Kosovska Mitrovica in an esteemed teacher's family. He finished the elementary school in Kosovska Mitrovica and the secondary shool in Belgrade. From 1941 to 1945 he was in a concentration camp, but at the end of the war he was a fighter in the national liberation army. He was holder of the Partizan Commemorative Medal 1941. Lj. Mihailovitsh entered the University School of Medicine in Belgrade in 1945/46 and finished the studies in 1951. During the studies he was one of the highly esteemed students and social-political workers. He was appointed physician at the Institute of Pathologic Physiology of the University School of Medicine in Belgrade in January 1952 where he started his research-work managed by professor Shahovitsh. At that time he spent half of his working hours at the Department of Neuro-psychiatry of the University School of Medicine in Belgrade managed by professor V. Vujitsh where he completed his knowledge in clinical neurology. In 1954 he was elected assistant lecturer, in 1958 senior lecturer, in 1964 associate professor, and in this rank he died. His remarkable working capability and brilliant intelligence became evident soon after the beginning of his duties at the Institute of Pathologic Physiology of the University School of Medine in Belgrade. In 1954 he went to Yale University in the USA for advanced, specialized training in the field of nerophysiology and neropathophysiology in the laboratory of professor P.J. Fulton. After return to the country in 1956 he became one of the leading research-workers, pedagogues and intellectuals. For his research-work in the field of pathologic physiology of epilepsy he got the Seventh July Award. On 21st March 1974 he was elected a corresponding member of the Serbian Academy of Science and Arts. The interest of professor Lj. Mihailovitsh was directed to several medical fields and therefore he became an eminent personality well known in the country and worldwide. He published 120 articles of which 365 are cited in scientific literature even today. The contribution of professor Mihailovitsh to the education of scientific personnel is great. In short, professor Ljubodrag Buba Mihailovitsh is one of the first ambassadors of Serbian neurophysiology. He succeeded not only in mastering the science in early youth, but also to contribute to its development. PMID- 12577682 TI - [Lifetime achievements of Dragan M. Dugalic. 1934-1992]. AB - Dragan M. Dugalitsh was born on 2nd August 1934 in Stari Trg in Kosovo in a working family. He finished the elementary school in his birthplace and the secondary school in Kosovska Mitrovica. He entered the University School of Medicine in Belgrade in 1954 and finished the studies in 1961 as one of the best students in his generation (average mark 9.22). During the studies D. Dugalitsh was very active in the students' organization, especially in professional and sport activities. He was educator first at the Chair of Chemistry and later at the Chair of Physiology. He was member of the Yugoslav Communist Party since 1954. He participated in three federal work drives. After having done the internship and national service he began to work at the Institute of Physiology of the University School of Medicine in Belgrade. However, in 1963 he changed the Institute for the First Department of Surgery managed by professor Ljubomir Rashovitsh. In 1968 he passed the examination in general surgery with excellent mark and in 1969 he was elected assistant lecturer at the Department of Surgery. His doctor's thesis was entitled "Experimental transplantation of lungs in dogs". Then, he was elected senior lecturer in 1975, associate professor in 1980 and professor of surgery in 1985. At the Department of Surgery he passed through all professional grades, from clinical physician and head of a ward to director of the Department of Surgery (1988-1992). He was also secretary of the Chair of Surgery (1984-1987) and associate dean of the University School of Medicine in Belgrade. Several times he received advanced training abroad. D. Dugalitsh was the visiting lecturer and surgeon in many towns in former Yugoslavia. He was member of many national and international surgical associations (SIC/ICC, CICD) and member of editorial boards of domestic and foreign medical journals (ACI, SA, AGEH). He was very active in the Serbian Medical Society, especially in the Section of surgery where he was also general secretary. He was rewarded by the Serbian Medical Society several times. In 1985 he was awarded the "Vishnjevski Medal" by the Moskow Vishnjevski Institute. In 1988 he became member of the Academy of Medical Sciences of the Serbian Medical Society. D. Dugalitsh was a workaholic, altruistic and self-denial person, especially devoted to an ill person. He was very respected by his colleagues and collaborators. He died on 3rd December 1992. PMID- 12577683 TI - [Lifetime achievements of Milan H. Djordjevic. 1933-1993]. AB - Professor Dr. Sci. Milan Djordjevitsh was an excellent cardiovascular surgeon and results of his professional and research work were well known to professional public in the country and over the world. He was born in Smederevska Palanka in 1933. He finished the University Schfool of Medicine in Belgrade in 1961 and specialized general surgery at the II Department of Surgery in Belgrade. His teacher, prof. dr. Vojislav Stojanovitsh, recognized his special interest for clinical and research work from the beginning. His doctor's thesis (1976) was "The Role of Veinous Valvula in Aortocoronographic Bypass During Increased Coronary Flow". In 1981 he formed a Bypass Centre of the Republic of Serbia which became later the Yugoslav European Reference Centre for pacemaker therapy. The same year (1981) he was rewarded "The October Award of the City of Belgrade" for science. He was the first to implant an implantable cardioverter defibrillator. This technique was carried out in Serbia only two months after the implantation of a similar system in Europe. Professor Djordjevitsh gave many lectures over the world: Detroit (1975), Huston (1975), Budapest (1978), Warsaw (1980), Stockholm (1982), Aman (1984), New-York (1988), Berlin (1987), Vienna (1998), Stockholm (1989), Moscow (1990). The professional and research work of professor Djordjevitsh in the field of electrostimulation of the heart was characterised by original ideas and permanent search for new possibilities, especially in the multidisciplinary field. Therefore, professor Djordjevitsh is considered to be the founder of modern clinical pacemaker therapy. He also practiced artistic and scientific photography. Since 1984 professor Djordjevitsh was member of the Nucleus of the European working group for heart electrostimulation and electrophysiology, and was elected in the presidency of the Nucleus; later he was its president. His interest was directed to the maturation of endocardial stimulation threshold; use of non-atrial sensors in the frequent adaptation of a permanent artificial conductor of cardiac rhythm-pacemaker to load; use of continuous electrostimulation of spinal cord in patients in terminal state of vasospastic and occlusive vascular diseases; study of the effort of electromagnetic fields on pacemaker inhibition and senzation; study of biologic synthetic grafts. As a man of high standing, professor Djordjevitsh was expert for cardiovascular diseases and modern technology in the World Health Organization since 1988. On the basis of his great professional reputation in the country and abroad, professor Djordjevitsh was elected a corresponding member of the Serbian Academy of Science and Arts. After a few scientific meetings excellently organized by professor Djordjevitsh in the country (Dubrovnik, 1988 and in 1990 under the auspices of NASPE), professor Djordjevitsh had to organize a European congress on Pacemaker Therapy and Electrophysiology in Belgrade in 1993. Unfortunately, political events and early death of professor Djordjevitsh were the reasons why the congress was not held as planned. Professor Djordjevic died in Paris, but was buried in Belgrade in January 1993. PMID- 12577684 TI - [Lifetime achievements of Bogomir B. Mrsulja. 1940-1944]. AB - Bogomir Mrshulja was born on September 18th, 1940 in Lendava. He finished both elementary school and high school in Belgrade and subsequently graduated from the School of Medicine, University of Belgrade in 1965. As an undergraduate he was an assistant at the Biochemistry Department and the subject became his profession for life. The University of Belgrade awarded him a scholarship and in 1968 he defended his MSc thesis entitled Glycogen of the Central Nervous System--Methods of Separation, Fraction Isolation, Regional Distribution and Variations in Different Levels of Deprivation of Paradoxical Sleep. Soon after, in 1970, he defended his PhD thesis, as well: Brain Glycogen and Deprivation of Paradoxical Sleep--Biochemical and Pharmacological Aspects of Glycogenolysis. In the period 1973-1975 he spent 18 months in the capacity of a visiting scientist in the Laboratory for Neuropathology and Neuroanatomic Sciences of the National Institute of Health in Bethesda (USA), and again, twelve more months in the same institution (Laboratory for Neurochemistry) in the capacity of a special expert of this institution (1983-84) where he was the chief investigator of neurochemical sequelae of experimental cerebral ischemia. He started his university career as an assistant professor in 1965; he was elected associated professor in 1970, promoted adjunct professor in 1978, while in 1984 he received the full professorship of biochemistry at the School of Medicine, University of Belgrade. In 1970 he was elected research associate of the "Sinisha Stankovitsh" Institute of Biological Research. He was professor of neurochemistry and Head of Department of Neurosciences of the Center for Multidisciplinary Studies, University of Belgrade. He was the Head of the Institute of Biochemistry and Head, Department of Biochemistry and Specialized Courses in Clinical Biochemistry at the School of Medicine, University of Belgrade. Owing to his original hypothesis on mechanisms of the occurrence of ischemic cerebral edema and possible therapeutic approach, Bogomir Mrshulja was frequently invited to lecture in university centers of USA, Canada, Japan, Germany and former USSR. He also participated at over 50 international congresses and symposia worldwide. In the mid seventies, Bogomir Mrshulja started more intensive studies of pathophysiological events in the course of cerebral ischemia and already with his early papers intrigued the professional circles which was reflected in an exceptionally high number of his citations in international scientific and referential literature, including the textbooks. The research activities of Bogomir Mrshulja was impressively fruitful: he published 183 papers in extenso, out of which 42 in books of international publishers and 6 invited review articles. In all these papers he illustrated extensive knowledge, maturity, originality, critical attitude, high level of creativity and, finally, courage to oppose the conventional principles. It should be pointed out that in his works Bogomir Mrshulja tended to draw conclusions that stimulated both himself and his colleagues, researchers in the related fields, insisting on extensive evaluation and testing of hypotheses, which is easily illustrated by the number of citations of his papers. According to the Science Citation Index, by 1995 he was cited 1700 times in the world literature, and in the period 1995-2002 he was cited 250 more times. Besides, he set up and headed the Laboratory for pathological neurochemistry at the School of Medicine in Belgrade that was the cradle for many outstanding researchers and lecturers that now work at the Schools of Medicine in Belgrade, Nish and Kragujevac, Military Medical Academy, and other institutions countrywide. He insisted on indispensable link between laboratory and clinical work, and, at least in the field of neurosciences, he is one of the pioneers of this kind of dialogue. In the year in which he left us for good (1994) scientific contribution of Bogomir Mrshulja was recognized by his election for the corresponding fellow of the Serbian Academy of Sciences and Arts. In the same year, he also received the October Award of the City of Belgrade for research. PMID- 12577685 TI - [Ten steps on the ladder of knowledge]. AB - The paper includes the report of the Editorial Board of the journal "Advances in Gerontology" dedicated to the issue of the 10th volume. There is an analysis of the topics and directions of published papers and their geographic and department distributions. PMID- 12577686 TI - [Achievements and prospects in the development of the St. Petersburg Institute of Bioregulation and Gerontology of the Northwestern Branch of the Russian Academy of Medical Sciences]. AB - The article presents the results of the 10-years' long activity of the St. Petersburg Institute of Bioregulation and Gerontology of the North-Western Branch of the Russian Academy of Medical Sciences founded to fulfill the basic and applied aims in the sphere of bioregulation and gerontology and to integrate with medical practice new methods of preventing premature ageing and age-related pathology by means of pharmaceuticals based upon peptide bioregulators. Among the Institute main achievements are the theoretical developments in the field of peptide regulation of ageing, creation of a new medical domain--bioregulation therapy, and foundation of the Russian school of biogerontology. PMID- 12577687 TI - [Aging of residents of St. Petersburg: current status and near-future prospects]. AB - Population ageing, an increase in the proportion of elderly, is a worldwide process. Having numerous economic, social and political consequences, population ageing has now risen to the top of developed nations' agendas. The evolution of ageing characteristics during 1970-2000 for Saint-Petersburg and their future trends have been considered. Comparisons with characteristics of ageing for Russia have been done. Special attention has been paid to gender disparities of ageing characteristics. PMID- 12577688 TI - [Effect of the season of birth on the mortality structure and life expectancy of residents of Kiev]. AB - Longevity was significantly associated with season of birth in 101,634 individuals who died in Kiev during the period 1990-2000. Longevity values were lowest for subjects born in April-July, and highest for individuals born at the end of the year. Minimum and maximum longevity by month of birth differed by 2.6 years in men and 2.3 years in women. It is shown that the persons born in the fourth quarter, died from accidents or suicide less often, than those born in the second quarter. That can testify their better social adaptability. Age of death of the persons who have died from age-related diseases (neoplastic and circulatory) depend on a season (month) of birth to the greatest degree. The obtained results testify that the early (pre- and/or postnatal) seasonal factors influence on age-related mortality and longevity of the people. PMID- 12577689 TI - Neurons, glia, and plasticity in normal brain aging. AB - Early manifestations of brain aging have received much less attention than the drastic degeneration of AD and MID. During nonpathological changes of normal aging, brain systems differ in the involvement of neuron loss: Spatial learning can become impaired without evidence for neuron loss, whereas eye-blink conditioning deficits are well correlated with Purkinje neuron loss. Glial activation, in particular the increased expression of GFAP, may be a factor in impaired synaptic plasticity. Lastly, I discuss how developmental variations in the numbers of Purkinje cells and ovarian oocytes can be factors in outcomes of aging that are not under strict genetic control. PMID- 12577690 TI - [Modern problems of the psychology of aging]. AB - Perspectives of aging psychology are discussed: behavioral, cognitive-behavioral, cognitive, psychodynamic, and contextual issues. Each of them has own principals and approaches; it were given examples of some theories. It is shown, that Russian authors are need to make their own priorities for effective researches and practical work with elderly. PMID- 12577691 TI - [Problems of the existence of "optimal" function survival functions in experiments of varying protein supplies to Mediterranean fruit flies]. AB - The paper is devoted to the discussion of the possibility of analyzing the changes in survival curves of Mediterranean Fruit Flies under different levels of reproduction by means of solving the optimization problem with some objective function. The investigation is carried out on the base of mathematical model with approximations resulting in the analysis of joint changes in survival functions and reproduction in flies subjected to different conditions of protein availability in food. The model describes possible mechanisms of evolutionary selection from the point of view of trade-off in average life span and reproduction. PMID- 12577692 TI - [Longevity of Drosophila after exposure to low doses of radiation and etoposide]. AB - The review of perennial researches of the authors on a problem of radiation induced aging is presented. From the point of view of radiogenetics it is represented perspective to investigate influence of an exogenous irradiating in small doses on fruit fly with defined mutation by phenotypes, that allows to repute a role of separate genes and mechanisms, controlled by them, in a determination of lifetime. At lines described by differences in pattern of mobile genetical elements, the different reaction to an irradiating explained from the point of view of features of the given mobile elements is found. The processing etoposide in concentration 5 microM on preimaginal stages of line with defects of a reparation (mei-41D5) and hypersensibility to an induction apoptosis (wg1-7, wg7L74, th1 [symbol: see text] th4) results in down-stroke of lifetime. The dominant effect of a gene mei-41D5 in a regulation of etoposide-induced change of lifetime is found. Is exhibited, that at lines with defects proapoptosis of genes reaper and Dcp1, the lifetime after influence, both ionizing irradiating, and inducer apoptosis etoposide is enlarged in comparison with the control. PMID- 12577693 TI - [Blood lipids, oxidative resistance of low density lipoproteins, concentration of fat-soluble antioxidants in older people and longevity in Novosibirsk]. AB - The aim of the study was to evaluate the blood lipid profile, oxidative antioxidative status of low density lipoproteins (LDL) in old people and long living people in Novosibirsk. Eighty nine persons from Novosibirsk Veteran House (23 men and 66 women) at the age of 83-105 (average 92.0 +/- 4.0 years old) were included into the study. They were divided into two groups. The group of old people included 12 person at the age of 75-89. The group of long-living people included 77 person at the age of 90-105. The control group consisted of 90 healthy subjects at the age of 36-59. TCH, CH-HDL and TG were measured using biochemical enzyme methods. LDL were isolated from serum by heparin-precipitation method. The initial level of peroxidation lipid (POL) products in LDL and oxidative resistance of precipitated LDL in vitro were evaluated. Concentrations of liposoluble antioxidants (a-tocopherol and retinol) in precipitated LDL were measured by spectro-fluorimetrical methods. The level of CH-HDL was low (< 40 mg/dl) in old and long-living people. In old people the initial level of POL products in LDL was higher in comparison with healthy subjects of control group (p < 0.0001) and long-living people (p < 0.01). In old people and long-living people the oxidative resistance of LDL in vitro was lower (p < 0.0001) in comparison with control group. Moreover, in long-living people the oxidative resistance of LDL in vitro was also lower (p < 0.01) in comparison with old people. The concentration of retinol in LDL was higher only in long-living people compared to the subjects of control group. The concentration of a-tocopherol in LDL was lower in old people and in long-living people (p < 0.05) in comparison with the subjects of control group. PMID- 12577694 TI - [Decrease of reactivity of cerebral arteries at late stages of life]. AB - The reactivity of cerebral arteries have been examined in three groups of patients (young, middle and adult ages). The medium meanings of the blood flow velocity (V) and the pulsativity index (Pi) were registrated in MCA and PCA under hypercapnia, hyperventilation and light stimulation. It was revealed that the reactivity of cerebral arteries in middle and especially in adult groups was decreased. PMID- 12577695 TI - [Geroprotective effect of thymalin and epithalamin]. AB - Researchers of the St. Petersburg Institute of Bioregulation and Gerontology of the North-Western Branch of the Russian Academy of Medical Sciences and the Research Institute of Gerontology of the Ukrainian Academy of Medical Sciences (Kiev) clinically assessed the geroprotective effects of thymic and epiphyseal peptide bioregulators (Thymalin and Epithalamin, correspondingly) in 266 elderly and older persons during 6-8 years (the bioregulators were applied for the first 2-3 years of observation). The obtained results convincingly confirmed the ability of the bioregulators to normalize the basic functions of the human organism, i.e. to improve the indices of the cardiovascular, endocrine, immune, and nervous systems, homeostasis, and metabolism. The restoration of homeostasis in the patients was accompanied by a 2.0-2.4-fold decrease in acute respiratory disease incidence, reduced incidence ischemic heart disease clinical manifestations, hypertension, deforming osteoarthrosis, and osteoporosis, as compared to the control group. Such a significant improvement in the somatic state of the peptide-treated patients corresponded to a decrease in their mortality rate during the observation period: 2.0-2.1-fold among the Thymalin treated patients, 1.6-1.8-fold--in the Epithalamin-treated group, and 2.5-fold- in the patients treated with Thymalin combined with Epithalamin, as compared to the control group. A separate group of patients was treated with Thymalin combined with Epithalamin annually for 6 years. We registered a 4.1-fold mortality decrease in this group as compared to the control level. The results of our research confirmed the conclusion on the high geroprotective efficacy of Thymalin and Epithalamin and the expediency of their application in medicine and social care as the means of health maintenance and age-related pathology prevention in persons over 60 years old enabling the prolongation of the active period of their lives. PMID- 12577696 TI - [Characteristics of effect of ultralow doses of vilon in organotypic culture of spleens from rats of various ages]. AB - There was studied the effect of vilon ultrasmall dose in the spleen organotypic tissue culture of rats of various age. The data obtained suggest that vilon ultrasmall dose (10(-9)-10(-13) M) exerted expressive effect on the explants development. This effect of effect of vilon ultrasmall dose can be used by the therapy, namely in the geriatric practice. PMID- 12577697 TI - [Functional morphology of an organotypic liver culture exposed to the peptide livagen]. AB - In this study the results of the tetrapeptide livagen effect on functional morphological characteristics have been produced. Under the data of the immunocytochemical and morphometrical analysis, tetrapeptide stimulates structural and functional homeostasis of cell populations in the liver culture. It was demonstrated that the main activity of investigated peptide had been directed to the stabilisation of morphological safety and reinforcement of the processes of cellular and intracellular forms of regeneration. PMID- 12577698 TI - [Effect of adjuvant magnesium therapy on the quality of life and emotional status of elderly patients with stable angina]. AB - The difficulties of treatment of the chronic coronar syndrome in the elderly patients depends on the severity of the disease and multimorbidity. Administration of magnesium preparations in these patients compensates the sytemic magnesium deficit. A number of physiological effects of magnesium allows to potentiate the antianginal effect of background drugs. The study of quality of life and of psychoemotional status in elderly patients with stable angina during the treatment with magnesium has been performed. It was observed an initial decrease of quality of life accompanied by anxiety and subdepressive emotional features. Adjuvant therapy with Magnerot and magnesium sulphate was followed by significant improvement in parameters of quality of life and by positive changes in psychoemotional status of patients. PMID- 12577699 TI - [Aging and carcinogenesis]. AB - The incidence of cancer increases with age in humans and in laboratory animals alike. There are different patterns of age-related distribution of tumors in different organs and tissues. Aging may increase of decrease the susceptibility of various tissues to initiation of carcinogenesis and usually facilitates promotion and progression of carcinogenesis. Aging may predispose to cancer by several mechanisms: 1) tissue accumulation of cells in late stages of carcinogenesis; 2) alterations in homeostasis, in particular, alterations in immune and endocrine system and 3) telomere instability linking aging and increased cancer risk. Increased susceptibility to the effects of tumor promoters is found both in aged animals and aged humans, as predicted by the multistage model of carcinogenesis. Available evidence supporting the relevance of replicative senescence of human cells and telomere biology to human cancer seems quite strong, however the evidence linking cellular senescence to human aging is controversial and required additional studies. Some common genetic processes (e.g. telomere dysfunction, changes in p53 and Rb activity and in DNA repair, accumulation of DNA lesions, genomic instability) play a critical role both in carcinogenesis and aging. Data on the acceleration of aging by carcinogenic agents as well as on increased cancer risk in patients with premature aging are critically discussed. In genetically modified mouse models (transgenic, knockout or mutant) characterized by the aging delay the incidence of tumors usually similar to those in controls, whereas the latent period of tumor development is increased. Practically all models of accelerated of aging in genetically modified animals show the increase in the incidence and the decline in the latency of tumors. Strategies for cancer prevention must include not only measures to minimize exposure to exogenous carcinogenic agents, but also measures to normalize the age-related alterations in internal milieu. Life-span prolonging drugs (geroprotectors) and genetic modifications may either postpone population aging and increase of tumor latency or decrease the mortality in long-living individuals in populations and inhibit carcinogenesis. At least some geroprotectors and modifications may increase the survival of a short-living individuals in populations but increase the incidence of malignancy. PMID- 12577700 TI - [Features of chemotherapy of malignant tumors in elderly patients]. AB - Elderly patients as a rule have modifications related to age (physiologic changes) and to other concomitant diseases (comorbidity). The glomerular filtration rate decreases with age. Before any chemotherapy is started creatinine clearance should be performed, and the doses of cytotoxic agents with active renal excretion should be modified according to creatinine clearance. Decreased liver blood flow may influence drug disposition. Some changes in liver function do occur with advanced age and may form the basis of some drug toxicities. The lean body mass in elderly people is characterized by an increase in fat, with a decrease in intracellular water, and this change, along with a decrease in serum albumin, can considerably modify drug pharmacokinetics. Decreasing in bone marrow reserve related to age increases the risk of myelotoxicity after cytotoxic therapy. Nevertheless chemotherapy may be tolerable and successful in elderly patients carefully selected. This article contains information about elderly cancer patients treated with cisplatin-based combinations (cisplatin + Vepesid + Nitrullyn, cisplatin + taxotere). There were no serious side effects. PMID- 12577702 TI - [Health promotion in Switzerland. Public health should carry more weight]. PMID- 12577701 TI - New challenges and old problems: end of life care and the dilemma of prognostic accuracy. AB - Dignity of the natural end of life for everybody is one of the new great challenges of medicine and social care for the beginning 21st century. However, many end of life care providing doctors are confused about how to categorize the help they give. One of the central problems is predicting the life expectancy of an individual patient. Difficulties in this field can become ethical dilemmas when physicians are obliged to predict accurately a patient's prognosis as the basis for a certain care strategy. Clinical estimation of the duration of life for patients with end of life cancer needs experience and training. Education programmes in the field should include this topic much more until now. Prognosis should be based more on proven indices and less on intuition. However, there is no doubt that daily clinical practice limits the use of highly sophisticated computer-based score models. Even maximal accuracy of prognosis will not exclude the risk of errors in a great part of patients. This limits their classification in care categories too strictly defined. Health care systems should avoid models for care with standards and budgets based on prognostic estimates and the medical community should avoid claim by disciplines of certain categories of patients defined by their prognoses. What we need is a network of assistance for incurable patients with single parts defined by patients needs and not by predicted life expectancy. Separating palliative and terminal care is artificial and often in contrast to the needs of the patients. PMID- 12577703 TI - [Working conditions. Stress--more than a social symptom]. PMID- 12577704 TI - ["One should deal with the origin of stress"]. PMID- 12577705 TI - [All fit. Empowerment for chronic patients]. PMID- 12577706 TI - [European network. Health in the routine school day]. PMID- 12577707 TI - [Oncology Congress. Quality of life despite cancer]. PMID- 12577708 TI - [Intermittent self catheterization. Compassionate nursing care is the beginning of success]. PMID- 12577709 TI - [A first class antique]. PMID- 12577710 TI - [Quality of life, a priority!]. PMID- 12577711 TI - [Promoting resources of daily life. Health: from ideas to action]. PMID- 12577712 TI - [Health at school--the Fribourg experience. "As good in my skin as in my class!"]. PMID- 12577713 TI - [The health promotion enterprise: example of l'Ecole La Source. Well being at work: an approach]. PMID- 12577714 TI - [Equilibrium atelier. Aplomb in the legs]. PMID- 12577715 TI - [Interdisciplinary collaboration. Nurse and doctor activities: sister and brother as Siamese twins?]. PMID- 12577716 TI - [Story writing in nursing care. Writing in all areas]. PMID- 12577717 TI - [out of scope]. PMID- 12577719 TI - [out of scope]. PMID- 12577718 TI - [out of scope]. PMID- 12577720 TI - [What is your diagnosis? Gynecomastia in Klinefelter syndrome (hypergonadotropic hypogonadism)]. PMID- 12577721 TI - [Interpretation of screening tests of hemostasis]. AB - The specialized hemostasis laboratory offers a huge amount of coagulation tests. However, the most important test for the general physician are the so-called coagulation screening tests. These tests include the prothrombin time, the activated partial thromboplastin time, the thrombin clotting time (often in combination with the reptilase time), fibrinogen concentration and the platelet count. Primary haemostasis is investigated by using the bleeding time or the platelet function analyzer (PFA-100). Depending on the results, in a second phase of investigation specific factor assays or other tests may be indicated. In the present work, we present the individual tests and discuss possible reasons for abnormal results. PMID- 12577722 TI - [Rapid progress in vascular surgery]. AB - Rapidly developed new technologies caused a lot of change in vascular surgery in the last few years. So today there are a lot of new therapeutic options, which we are using with impressive speed directly in humanbeing. We look at new treatment modalities in the surgery of the carotid artery and the abdominal aortic aneurysm. PMID- 12577723 TI - [The brain in rapid technological progress--epileptology for the general practitioner]. AB - Some important aspects of the therapy of epilepsies are treated. The practitioner is often confronted with an acute epileptic attack: guidelines for the management are given. As recurrences are not too frequent, treatment usually is only begun after a second seizure and the diagnosis must be confirmed. Compliance is of utmost importance, it can be improved by good advice and guidance. Recommendations for starting and stopping treatment are given. Pregnancy is a particular challenge. Most women can give birth by natural ways and have healthy children. Sudden unexpected death is a rare but important problem, mostly in connection with an attack. PMID- 12577724 TI - [Rational assessment and therapy of diarrhea]. AB - Patient history, physical examination and basic laboratory tests often allow the clinician to suspect one of the numerous aetiologies of diarrhea and consequently establish a rapid diagnosis. Otherwise, the workup of diarrhea, especially when chronic, may represent a challenge. In this brief tutorial article a practical algorithm is proposed to guide the clinician through the initial diagnostic steps as well as therapeutic trials. PMID- 12577725 TI - [Running from myocardial infarct and the cardiologist. Jogging instead of stenting]. PMID- 12577726 TI - [Also true in coronary heart disease: the well trained patient lives longer]. PMID- 12577727 TI - [Infarct risk increases in sports. Only training can prevent it]. PMID- 12577728 TI - [Even in advanced age: statins prevent infarct]. PMID- 12577729 TI - [Statins for all seniors?]. PMID- 12577730 TI - [Young and already considerably overweight. Children's hearts are not designed for this stress]. PMID- 12577731 TI - [Prevention of thrombosis in long distance flights. Support or injection?]. PMID- 12577732 TI - [Can new therapy concepts in oncology still be realized? Earlier death with standard therapy]. PMID- 12577733 TI - [Rethinking in oncology. More quality of life for patients by information? (interview by Christine Vetter)]. PMID- 12577734 TI - [Allergy test in general practice? Patients know who is more competent]. PMID- 12577735 TI - [Subungual warts on the toes. How can I get rid of them again?]. PMID- 12577736 TI - [Hearing loss due to leisure time noise is on the rise. The ear also needs a rest period]. AB - Noise during leisure time is increasingly leading to temporary and permanent hearing loss, with most victims being adolescents and young adults. Particularly harmful is permanent exposure to noise, since the highly sensitive ear is not designed to tolerate it. On average, youngsters aged between 14 and 20 years spend 3 hours and more listening to music. In discos, concerts in particular, as well as listening to music via headphones, sound levels of 100 dB are regularly reached and exceeded, and levels of 90 dB are already considered to represent a definite risk. Noise is, however, experienced as offensive and painful at levels of about 120-140 dB, and this leads to the danger of its harmful effects being underestimated. Treatment of hearing loss depends on whether it is acute or chronic and comprises infusion therapy, stress reduction, acoustic rest or interposed periods free of noise and, where indicated the fitting of a suitable hearing aid. In view of the immense significance of hearing impairment for the economy, public health, and society at large, its prevention--not only by protective, but also preventive, measures--warrants greater attention. PMID- 12577737 TI - [40% of hearing aids are not worn. Making sure the device doesn't wind up in the drawer!]. AB - Early diagnosis of a hearing impairment, follow-up and timely effective intervention are the aims of optimal management of the hard-of-hearing patient. Today, every fifth adult has a hearing problem. This means that the general practitioner not only faces a diagnostic challenge, but also needs to concern himself with the prevention of a hearing loss or its further progression. In the event of a hearing aid being required, the individual needs of the patient must be taken into account. Currently available are behind-the-ear and in-the-ear, both as digital and analog devices. The most expensive, smallest and most modern device is not necessarily the best option for the individual patient, and fitting should be a cooperative effort on the part of the ENT physician and the hearing aid specialist. Required is a specific history, comprehensive counseling of the patient, a phase of testing at home and, where necessary, concomitant acoustic training. The German Society for the Hard-of-Hearing has advice centres in many towns and cities that provide information and counseling on rehabilitation and the legal rights of the deaf. PMID- 12577738 TI - [Screening methods for detecting congenital hearing disorders. Should the newborn infant be tested?]. PMID- 12577739 TI - [References for communication, social contacts and career choice. Making life easier for the hearing impaired patient]. PMID- 12577740 TI - [Hyperkalemia in an elderly diabetic patient. Is it caused by "kidney flushing"?]. PMID- 12577741 TI - [Careful assessment is advisable. Sleep apnea: risks in daily living increase, vitality decreases]. PMID- 12577742 TI - [Often, general measures are adequate. The sleep laboratory is not always necessary]. PMID- 12577743 TI - [Crossectomy--exhairesis--stripping--laser therapy. How even refractory varicose veins respond to treatment]. AB - Chronic venous insufficiency affects more than 50% of the German population. Major factors involved in its development are age, family disposition, female sex and an occupation involving much standing. Together with the clinical presentation, Doppler and duplex ultrasonography in particular enable a reliable pre-operative diagnosis, and deep venous thrombosis can also be definitively excluded. Indications for surgical treatment are in particular varicosis of the greater and lesser saphenous vein and perforating vein insufficiency. Commonly used procedures are crossectomy, restrictive stripping of pathological vein segments, resection of varicose side branches, and the endoscopic discission of perforating veins. Recent developments are deep-freezing and extraction of the vein and endovenous laser treatment (EVLT), requiring only tiny incisions. In most cases, these interventions can be performed on an outpatient basis. PMID- 12577744 TI - [Simple, safe and effective. New protease inhibitor means progress]. PMID- 12577746 TI - [Hepatic encephalopathy. Flickering light exposes subclinical forms]. PMID- 12577745 TI - [D-hormone reduces rate of falls. In osteoporosis consider more than bone density]. PMID- 12577747 TI - [Beta blocker enhances NO liberation in the endothelium. Vascular tone long-term improvement]. PMID- 12577748 TI - [Fear of the injection must not be an argument. Every second type 2 diabetic patient needs insulin]. PMID- 12577749 TI - [Despite improved healing chances with pegylated interferon, hepatitis C is frequently not optimally treated]. PMID- 12577750 TI - [Patient with chronic backache. Disabled or muscle spasm patient?]. PMID- 12577751 TI - [The greatest number of members of the German Pharmaceutical Society?]. PMID- 12577752 TI - [Living collegenin and collagen]. PMID- 12577753 TI - [Dangerous interaction of St. John's wort with chemotherapy?]. PMID- 12577754 TI - [The mechanism of action of organic nitrates]. PMID- 12577755 TI - [From teprotide to captopril. Rational design of ACE inhibitors]. PMID- 12577756 TI - [Intervention in the renin-angiotensin and kallikrein-kinin systems. Pharmacologic basis of the action of ACE inhibitors]. PMID- 12577757 TI - [Now well established! The importance of ACE inhibitors in high blood pressure therapy]. PMID- 12577758 TI - [The importance of ACE inhibitors in the treatment of cardiac insufficiency]. PMID- 12577759 TI - [New perspectives for patients at risk for atherosclerosis. The importance of ACE inhibitors in cardiovascular prevention]. PMID- 12577760 TI - [In arterial hypertension: ACE inhibitors and combination therapy]. PMID- 12577761 TI - [Management of high blood pressure and cardiac insufficiency. New drugs and therapeutic concepts]. PMID- 12577762 TI - [To what should we pay attention? Pharmaceutic care for patients with cardiac insufficiency]. PMID- 12577763 TI - [Depression also effects the body]. PMID- 12577764 TI - [Adalimumab slows the progression of rheumatoid arthritis]. PMID- 12577765 TI - [The challenge of Alzheimer dementia]. PMID- 12577766 TI - [Immunodeficiency in adult patients as the first sign of a primary underlying condition]. AB - Three male patients, aged 56, 40 and 52 years, presented with recurrent infections and clinically suspected immunodeficiency, which was confirmed by the presence of hypogammaglobulinaemia. In one patient, no obvious underlying disease was identifiable, and primary immunodeficiency syndromes were considered in the differential diagnosis. A thorough diagnostic work-up revealed the presence of non-Hodgkin lymphoma in an iliac crest biopsy specimen. The second patient was found to have a thymoma. In the third patient, the immunodeficiency was the key to the ultimate diagnosis of multiple myeloma, which was still asymptomatic at that stage. Establishing the underlying condition had important therapeutic and prognostic consequences in these patients. PMID- 12577768 TI - ['Proteomics': the mapping of all human proteins]. AB - The genomes of many organisms, including humans, are now largely known. In the wake of this there is a need to identify and measure all proteins that are encoded by the genome (proteomics). This need leads to turbulent developments in the area of analytical techniques, such as two-dimensional electrophoresis, mass spectrometry, and protein chips. The rapidity of advancements justifies the expectation that in the next 5-10 years it will indeed become possible to determine the proteome of an organism or its components such as plasma, serum, or tissues. In combination with information on initiation and progress of disease, proteomics will contribute to improving health and to better primary and secondary prevention. PMID- 12577767 TI - [Treatment of hypertension: angiotensin-II antagonists potentially better than beta-blockers in the occurrence of cardiovascular and cerebrovascular damage; the LIFE study in perspective]. AB - Current clinical guidelines state that only beta-blockers, diuretics, calcium channel antagonists and ACE inhibitors should be used for initial pharmacotherapy for uncomplicated hypertension. On basis of experience, efficacy and costs beta blockers and diuretics are first choice. However, the importance of the renin angiotensin-aldosterone-system in the pathophysiology of hypertensive end-organ damage is increasingly recognised nowadays, and modulation of this system may therefore exert favourable effects on cardiovascular and cerebrovascular complications. In the LIFE study, a recently published double-blinded, randomised trial, the angiotensin-II receptor (A-II) antagonist losartan was compared with the beta-blocker atenolol in patients with essential hypertension and left ventricular hypertrophy (LVH). Patients randomised to the A-II antagonist suffered statistically significantly fewer clinical end-points, specifically fewer cerebrovascular accidents, whereas both treatments resulted in a similar decrease in blood pressure. In the subset of diabetic patients, the use of the A II antagonist yielded an even more favourable outcome. In our opinion, it should now also be permitted to prescribe A-II antagonists as initial pharmacotherapy for patients with uncomplicated essential hypertension. It might be considered to prescribe A-II antagonists as preferred treatment for patients with essential hypertension and known LVH, especially in diabetic patients. PMID- 12577769 TI - [Chronic lymphocytic leukemia: high time for a risk-adapted approach]. AB - B cell chronic lymphocytic leukaemia (CLL) is the most common leukaemia in the Western world. It is wrongly considered to be an indolent disease: even patients with Binet stage A can have disease-related morbidity, necessitating treatment in about 50% of them, and over 25% of these patients will die of CLL-related causes. It was recently discovered that there are in fact 2 subtypes of CLL: a pre germinal centre type. Characterised by unmutated genes which code for the variable parts of immunoglobulin chains (IgV), and a post-germinal centre variant, characterised by IgV somatic mutations. This IgV-gene mutational status as well as surface expression of CD38 and characteristic cytogenic abnormalities has recently been shown to be powerful prognostic factors. Furthermore, over the past few years new treatment modalities have been developed including purine analogues, immunotherapy with monoclonal antibodies and stem cell transplantation. Therefore it is time to substitute the present policy used for the vast majority of patients (i.e. an expectant approach, if needs be followed by initial treatment with chlorambucil) with an approach based on the patient's individual risk profile. This should preferably be done within the framework of clinical trials. PMID- 12577770 TI - [Admission and discharge criteria for intensive care departments]. AB - Admission and discharge criteria for intensive care departments have been drawn up in order to optimise the use of scarce and costly intensive care facilities. Every patient who could benefit from admission must be assessed by the intensive care specialist beforehand. Admission is indicated for patients with disrupted vital functions in whom recovery of dysfunctioning or failing organ systems is expected, patients who will act as organ donors and patients who undergo diagnostic investigations associated with a high risk of vital complications. Frequent assessment (several times per day) of the 'indication to stay' is indicated in the case of many patients in order to maximise the admission capacity. Discharge from the intensive care department is indicated if the vital functions are stable without life support and no longer require monitoring or treatment, if nursing the patient in the ward is possible, if continuation of the medical treatment is no longer worthwhile, if the patient no longer consents to the treatment and if the benefit of a treatment no longer outweights its negative effects. PMID- 12577771 TI - [Diagnostic image (122). A boy with serious rectal bleeding. Meckel's diverticulum]. AB - An 11-year-old boy presenting with serious rectal bleeding had a negative Meckel scintigraphy. On a barium small bowel followthrough, a small accumulation of contrast was seen outside the contour of the ileum, caused by a Meckel's diverticulum after all. PMID- 12577772 TI - [Favorable results with duplex-guided compression sclerotherapy for varices of the small saphenous vein; a retrospective study]. AB - OBJECTIVE: Assessment of the efficacy of duplex-guided compression sclerotherapy in patients with varices of the small saphenous vein (SSV). DESIGN: Retrospective. METHOD: Data were collected from 109 patients (14% male and 86% female; average age 51.4 years (SD: 10.6)) with 121 SSV varices which were sclerosed under duplex guidance with polidocanol 3% during the period 1 December 1998-31 May 2001 in the Dermatology department at Maastricht University Hospital, the Netherlands. After-care consisted of a compression bandage for 1 week and a therapeutic elastic stocking for 6 weeks. Check-ups took place after 3-6 months and after 12 months. Sclerosis was repeated if the procedure had failed. Success was defined as occlusion or by the absence of reflux and a reduction in the complaints. RESULTS: After 3-6 months the treatment was successful in 88/113 varices (78%) for which data were available, and after 12 months treatment was successful in 46/57 (81%). CONCLUSION: Sclerosis of the SSV under duplex guidance, with repeat treatment if required, gave favourable results. PMID- 12577773 TI - [The clinical diagnosis 'wound botulism' in an injecting drug addict]. AB - A 38-year-old subcutaneous injecting heroin addict had subacute blurred vision, dysarthria and dysphagia. The next day she could not swallow or speak and developed weakness of all limbs and respiratory failure. Electromyography showed abnormalities compatible with a presynaptic neuromuscular transmission deficit, which supported the diagnosis of botulism. The point of entry was probably a skin abscess due to injections. Treatment with antitoxin and antibiotics resulted in a favourable recovery. Wound botulism is caused by local production of toxin by Clostridium botulinum after wound infection. Although it is a rare variant of botulism, it is increasingly being reported in drug users who inject subcutaneously. PMID- 12577774 TI - [More care for a limited budget; a case for a better use of the efficiency criteria]. PMID- 12577775 TI - [Increase in rate of resistance to fusidic acid among Staphylococcus aureus isolates from patients admitted with atopic dermatitis]. PMID- 12577776 TI - [Misleading advertising claims about the indication of doxazosin (Cardura) for type 2 diabetes mellitus and hypertension]. PMID- 12577777 TI - [Cephalic vein access for implantable venous access devices. Technique and long term follow-up]. AB - OBJECTIVE: To evaluate 1) the advantages and drawbacks of the cephalic vein access for totally implantable venous access devices (TIVAD); 2) the complications of such devices at long-term follow-up. PATIENTS AND METHODS: A retrospective study of a series of 107 cephalic vein accesses attempted on 105 patients from January 1997 to December 2001, in our department. They were 87 men and 18 women, aged from 25 to 85 years (mean: 59). The authors evaluated in each case the surgical technique, the preoperative incidents, the side of surgery, and the short-term and long-term complications. The total follow-up reached 41 284 days. RESULTS: The cephalic vein access attempt failed in 12 cases (11%). These failures were due to intravenous blockade of the flexible catheter or to misroutine of the catheter in 5 cases and to a too thin vein in 7 cases. There was no statistically significant difference between the right and left sides. There was no technique-related complication. There were 18 postoperative complications (19%) out of the 95 devices that were implanted: 6 venous thrombosis, 7 infections, 4 catheter disconnections including 3 intra-cardiac migrations, 1 catheter occlusion. In 12 of these cases, the whole device had to be withdrawn. In the whole series, the devices were withdrawn at mean of 435 days after implantation. CONCLUSION: The use of a TIVAD is necessary when a chemotherapy must be delivered for treatment of a cancer. Cephalic vein access appears to be easy, quick and relatively safe. Postoperatively, serious complications can be encountered like with any type of TIVAD. These complications must be well-known if one wants to manage them well. PMID- 12577778 TI - [Pneumoparotid: a case report and review of the literature]. AB - This article reports a case of peneumoparotid in a ten-year-old girl. This pathology is caused by insufflation of air into the acini of the parotid gland via Stensen's duct. A review of the literature shows that it has many aetiologies but more frequently it presents as an occupational hazard among glassblowers and wind instrumentalists. It may also occur by auto-insufflation in adolescence who often have psychological problems. The diagnosis is made from the history and imaging. The treatment is aetiologic and symptomatic. PMID- 12577779 TI - [Value of of fine-needle aspiration cytology and MRI in parotid gland masses]. AB - INTRODUCTION: The objective of our study was to discuss the valve of fine-needle aspiration cytology (FNAC) and magnetic resonance imaging (MRI) in the diagnosis and treatment of parotid gland masses. MATERIALS AND METHODS: Forty patients were included in the prospective study. They had undergone clinical examination, FNAC and MRI before parotidectomy. The results of these examinations were compared with the corresponding histopathological diagnosis. RESULTS: When it is positive, FNAC is a good examination of malignant tumours (sensitivity 67%, specificity 79%, positive predictive value 86%, negative predictive value 100%). The MRI allows a good assessment of the tumoural mass and its anatomical relationships (sensitivity 55%, specificity 86%, positive predictive value 89%, negative predictive value 75%). If the T2 sequence shows reduced density (p < 0.05) or in case of bad limitation (p = 0.004), a malignant character is strongly suspected. CONCLUSION: In cases of parotid gland mass, where surgical intervention is necessary, there is no need of special investigations: however FNAC and MRI allow us to anticipate what operation will be required. PMID- 12577780 TI - [Parathyroid cervical hematoma: a case report]. AB - Haemorrhagic cysts of the parathyroid gland are rare. Our case report concerns a presentation with a constrictive cervical haematoma, confirmed by ultrasound guided aspiration. The haematoma, rapidly recurred, indicating exploration of the neck and haemostasis. Histological examination of excised tissue showed the haemorrhagic nature of the cyst. PMID- 12577781 TI - [Value of interventional video-endoscopy in the assessment of cancer of the upper airways and digestive system]. AB - INTRODUCTION: The aim of our study was to evaluate the video fiberoscope with operator canal (FOC) in he diagnosis of head and neck carcinoma compared to direct laryngoscopy (DL). METHODOLOGY: From August 2000 to May 2001, 82 patients were included in a prospective study. They were examined for pharyngolaryngeal cancer with the FOC, DL and a CAT-SCAN. The compared elements were the visualization of the different regions of the pharyngolarynx. A three dimensional description of the lesion, the presence of secondary localisations as well as the anatomo pathological results were obtained with FOC and DL. RESULTS: Although the pharyngolarynx is less well visualized in FOC than in DL (p = 0.04), no statistically significant difference was found between the two methods in terms of visual diagnosis and assessment of the extent of the lesion. The sensitivity of the biopsies made with FOC is lower than those performed with DL (66% FOC, 97% DL). Moreover, we have noted that FOC is highly reliant on the experience and ability of the surgeon. CONCLUSION: DL remains the most reliable technique for the exploration and diagnosis of the pharyngolarynx. If it is not possible to perform a DL or if it is not indicated (trismus, contra-indication to general anaesthesia) FOC should be considered as the examination of choice. PMID- 12577782 TI - [Laryngeal tuberculosis: report of a case]. AB - The larynx as a site of tuberculosis represents less than 1% of the total of this disease. Primary tuberculous laryngitis is even more rare. The authors report a case of an 89 year-old man presenting with swallowing disorders (dysphagia and overspill) consulting for suspected laryngeal carcinoma. Finally, the diagnosis was laryngeal tuberculosis. The outcome was favourable with appropriate treatment, obtaining complete healing without any after-effects. The diagnosis of tuberculosis must cross the clinician's mind, even when there are no general symptoms. PMID- 12577783 TI - Arteriovenous haemangioma of the pterygoid process. Report of a case. AB - Arteriovenous haemangioma is a benign tumor which in ear, nose and throat practice is generally found in the soft tissues of the oral cavity. Bone involvement is very uncommon. These lesions typically manifest themselves in infancy, with a tendency towards spontaneous resolution. The management options comprise simple observation, embolization or surgical removal--depending on the location of the tumour and its symptoms. A correct diagnosis is essential to ensure the adoption of adequate measures during surgery. The present case describes an exceptional location of arteriovenous hemangioma of the pterygoid process, and discusses the associated diagnostic and treatment problems. PMID- 12577784 TI - [Nasal involvement in Crohn's disease: report of a case and diagnostic difficulties of systemic diseases affecting the nose]. AB - The authors describe the case of a 32 years old man with Crohn's disease who has developped sinonasal and tracheal localizations. The review of the literature, quite poor, allows to find some cases of nasal involvement in Crohn's disease and few associations between this illness and others systemic diseases as Wegener's disease or relapsing polychondritis. Among these diseases, we can find atypical conditions mixing different features of the most classical systemic diseases, which are called "overlap syndromes". The case we report illustrate these questions relative to the diagnosis. To conclude, it may be important to point at the difficulties of the anatomopathological diagnosis and the interest of the iterative biopsies. Regular follow-up can help us to identify systemic disease with the research of new extra-sinonasal symptoms. PMID- 12577785 TI - Functional septal surgery: computer simulated surgery. AB - Septal surgery is one of the most commonly performed nasal surgeries. It aims at restoring nasal patency and symmetrical air passages. However classical surgery may narrow the originally wider side necessitating additional surgery at the level of the turbinates. This study was performed to demonstrate that properly targeted and tailored surgery with limited interference at the level of specific areas may not only be enough but it may also more efficacious than more extensive surgery. The coronal CT scans of 16 patients with septal deviation were transferred to a computer and the cross-sectional areas (CSA) of both passages calculated before and after various simulations of septal surgery. In 8/16 cases the more limited surgery resulted in a wider total CSA and also more symmetrical right and left passages. This is more physiological both in terms of function and subjective feeling by the patients. We suggest that for all patients planned to undergo septal surgery a pre-operative coronal CT should be studied and the surgery planned to achieve the most optimal result rather than remove indiscriminately all deviated elements. PMID- 12577786 TI - [Management of a giant ameloblastoma at the Butare University Hospital Center (Rwanda)]. AB - The management of ameloblastoma is nowadays well codified, requiring surgical treatment. If clinical and radiological aspects could evocate diagnosis, only histological analysis proves it, and permits to differ ameloblastoma from other facial bone tumors. Due to difficulties in Rwanda to obtain preoperative X ray computed tomography, the diagnosis and the management are first based on the clinic features. PMID- 12577787 TI - [A case of partial hearing recovery after ablation of cerebello-pontine angle neurinoma]. AB - A case of a 44 year-old woman farmer with a total deafness and lack of vestibular function on the left side operated upon the cerebello-pontine cystic neuroma 2.5 x 2.0 x 1.5 cm large via the retromastoidal craniectomy is presented. Despite of the negative audiological tests' results the cochlear nerve flatten on the tumor was cautiously separated. Some time after surgery, she observed "a partial recovery of hearing" in the operated ear confirmed by the tonal (mean 0.5-2 kHz 36 dBHL) vocal SRT 70 dB), BERA and EOAEs tests. The paralysis of the left vestibular function did not recover. In consecutive audiogram a notch at 6 kHz 15 dB deep appeared in the operated ear and one can presume an increased vulnerability of this ear to the noise of the agricultural machines used by then by the patient. PMID- 12577789 TI - [Collecting points with the MMP]. PMID- 12577788 TI - [Meetings of the IPSEN Foundation: an appointment between the cochlea and the retina]. PMID- 12577790 TI - [Undesired drug reactions: causality assessment]. PMID- 12577791 TI - [Prolonging the QT time with drugs]. PMID- 12577792 TI - [Biological weapons. II. Smallpox, botulinum toxin, hemorrhagic fever viruses]. PMID- 12577793 TI - [Drugs in pregnancy. Treatment of duodenal ulcer]. PMID- 12577794 TI - [Report of 243 vertical mammoplasties for very large, heavy breasts and/or severe ptosis. Analysis of the result and technical]. AB - Vertical mammaplasties for very large breasts and/or severe ptosis were evaluated in 124 patients who underwent operation in our unit between September 1993 and June 2001. In 119 cases it was reduction mammaplasty and in 5 cases unilateral symmetrization after contralateral reconstruction. The mean age was 36 years (13 62 years). Among inclusion criteriae, we choose the resection weight > or = 700 g and/or a ptosis > or = 30 cm. We report a few technical modifications of the initial Lejour vertical technique. About hypertrophic breasts (179 breasts): mean resection weight was 905 g (710-1750 g), mean ptosis was 33.5 cm (30-42 cm). The following complications were noted: 3 haematomas with only one evacuation on the same day, no seroma, 4 infections controlled by local treatment, 44 wound dehiscence or delayed skin healing (> 30 d), 8 partial necrosis, 12 secondary correction under local anaesthesia, 1 cheloid scar of the areolar complex. About breasts with severe ptosis (64 breasts): the mean value was 31.8 cm (30 cm). Complications were as follow: 2 haematoma, 0 seroma, 16 healing delay, 3 partial necrosis, 6 secondary correction under local anaesthesia. The mean procedure time was 94 mn. The mean stay in hospital was 2.8 days, same as the suction drains. The vertical mammaplasty is a fast technique, giving good results. Scarring amount is reduced. Breasts are well projected, well shaped and remain stable in time. Their base can be modified. We think this technique can safely be applied to large breasts and/or severe ptosis. PMID- 12577795 TI - [Our experience with tissue expansion on a series of 50 cases in Vietnam]. AB - The authors reviewed their experience with tissue expansion from July of 1995 to December of 1999 at Hanoi Plastic Surgery Center. A total of 75 tissue expanders of various sizes were placed in 50 consecutive patients (16 men and 34 women) for the reconstruction of secondary defects (burn scars, skin graft scars, hypertrophic scars, keloids, capillary hemangioma, congenital nevi and micotia). The average age of patients was 21 years. The tissue expansion protocol was used in clinical as well as common principle. The main technical details are modified in this procedure by the authors: type of intralesional incision for expander insertion, closing of wound incision by three layers, evacuation of the liquid in the prosthesis pocket, injection with antibiotic solution and expanded flap capsulectomy. The complications rate was 10.6% (8 complications in 75 expanders). The most common complications consisted of infection, hematoma, exposure of valve, dehiscence of incision, necrosis of the distal expanded flap. The overall failure rate was 8%. Thus our modified surgical details allowed us to decrease the major complications and to achieve the best possible functional and aesthetic results. PMID- 12577796 TI - [Rhinoplasty in the immediate assumption of responsibility of nasal burns. Technical note and report of two cases]. AB - We report the use of a traditional technique of rhinoplasty in the immediate assumption of responsibility of deep burn of the nasal edge which cannot be grafted. The limit of the realization of this gesture lies in the fact that it must exist nasal morphological characteristics at the patient. The two cases presented had an hypertrophic nose with osseous and/or cartilaginous bump. The reduction height of the osseo-cartilaginous frame allows closing the defect without cutaneous tension. One carries out a true excision-joining of the burn of the nasal dorsum. We note at one year of retreat, the discretion of the scar located on the centre line of the nose, the absence of after-effects as well as the cosmetic satisfaction of the patients with respect to the modifications morphological their noses. PMID- 12577797 TI - [Post-inflammatory cutis laxa. A case report]. AB - Cutis laxa (CL) is a rare disorder in which the skin hangs in loose fields, so that affected individuals appear to be prematurely aged. The changes are more evident when effect the face with a prematurely-aged appearance. The acute form follows an inflammatory skin lesions. Its aetiology is not well known. We report a case of a head acute CL of a 17 years patient, secondary to a generalized skin eruption. The prematurely-aged appearance concern especially forehead, ear lobes and nasolabial folds. A two stages surgical treatment has associated: a forehead lifting associated to a Coleman lipofilling and an ear lobe reduction, a secondary Coleman lipofilling of the nasolabial folds. The skin biopsy confirm the diagnosis. The correction is stable and satisfactory after one year. The confrontation of our findings to those previously described confirm complexity of diagnosis and histological observations of this rare disorder. PMID- 12577798 TI - [Vascular leiomyoma of the hand. Report of a case and review of literature concerning a rare pathology]. AB - Vascular leiomyoma is a benign painful tumour that occurs most frequently in the lower extremity. We report an uncommon case in the hand. Only surgical removal of the tumour allows establishing the diagnosis histologically. A review of the literature confirms the rarity of the tumour in the hand. The purpose of this case report is furthermore to consider the differential diagnosis in painful masses of the hand. PMID- 12577799 TI - [Should residents operate?]. AB - Surgical training requires that residents have the opportunity to operate. This tradition has now become complicated for teaching surgeons due to work surcharge, to the objective of reducing health costs, and to medical lawsuits. We support the necessity for residents to operate because academic hospitals have a moral and legal obligation to train future surgeons. This objective should be achieved without impairing the quality of care provided, and this must be explained to patients. More widely, the management of surgical training for residents should be rethought to maintain efficiency. PMID- 12577800 TI - [The 16th Congress of the l'ISAPS, May 25-28, 2002, Istanbul. What is in store for ISAPS: the end of "cultural exception"?]. PMID- 12577801 TI - [Involvement of Polish enterprises in workplace health promotion. Trends observed in years 1998-2001]. AB - The author presents voluntary activities aimed at improving the workers' health in medium-sized and large enterprises in Poland, as well as the dynamic of changes in this area. The data were collected from enterprises chosen at random in 1998 (147 enterprises), 2000 (755) and 2001 (215). At present, over 60% of enterprises declare that they do more for their workers' health than it is required by legal regulations. Since 1998, the number of such declarations has been constantly growing. It is observed that the larger enterprise and the better economic situation, the more frequent health promotion activities. Two areas of activities are most commonly carried out: (1) medical services, e.g. vaccinations, extra diagnostic examinations and therapeutical services; and (2) investments in the work environment, e.g. ergonomics, improvement of interior designs. Unfortunately, promotion of healthy lifestyles, e.g. non-smoking campaigns or stress management are definitely less frequent. Although a growing number of enterprises becomes involved in health promotion, the range of their activities is diminishing. The programs aimed at promoting health conductive behaviors among workers are in particular given up. As far as smoking cessation is considered, the enterprises disseminate materials on the adverse effects of smoking or related ill-health and punish smokers for breaking smoking bans instead of helping their workers and teach them how to quit smoking or support those who want to give up this habit. In the years 2000 and 2001, the educational and supporting activities decreased whereas the punitive policy was on the rise. PMID- 12577802 TI - [Workplace health promotion program quality evaluation questionnaire]. AB - There are substantial reasons for undertaking the evaluation of workplace health promotion (WHP) programs. The most important ones are: (a) the need to guarantee a competent organization of programs; (b) the feasibility to evaluate WHP programs, which take the form of medical services offered to employers on the health service market; (c) the need to disseminate the idea and support the workplace health promotion lobby; and (d) the use of evaluation as a tool for analyzing WHP promulgation process. The implementation of the scientifically based evaluation of health promotion programs carried out in regularly functioning enterprises is very limited, hence the increasing interest in procedures of quality assessment already known and accepted by enterprises. This approach is recommended by the World Health Organization (WHO). In the European Union countries, a questionnaire on the workplace health promotion program evaluation is promulgated within the European Network for Workplace Health Promotion. The National Center for Workplace Health Promotion that coordinates the activities of the National Network for Workplace Health Promotion Centers in Poland has developed a Polish version of the questionnaire adopted to local conditions. The author presents the criteria and the most essential solutions that provided the ground for designing the questionnaire to be used by the organizers of WHP programs in enterprises for self-assessment of their own activities. It reflects the views on the strategy of the WHP evaluation presented by the National Center, and contains a simplified procedure that does not involve control groups so difficult to gather under the concept of "health promoting enterprise". The idea of control groups is usually misunderstood and disapproved by the management of enterprises involved in the implementation of WHP programs. PMID- 12577803 TI - [Occupational diseases among personnel of Polish hospitals, 2001]. AB - The paper presents the data provided by the Central Register of Occupational Diseases in Poland on the compensated occupational diseases among hospital personnel, registered in 2001. The trends in the incidence of occupational diseases in this population over the period 1994-2001 are also discussed. In total, 394 new cases of occupational diseases among the hospital personnel were registered in 2001, which makes up 52.1% of all cases recorded under the "Health and social work" section of occupational activities, according to the Nomenclature des Activities de Communite Europeene. Most of these cases were found among nurses (47%), followed by physicians (15%), laboratory analysts (11.5%), orderlies (11%), and dentists (3%) and referred mainly to females (84.8%). Contagious and invasive diseases prevailed, constituting 73.9% of all cases. Viral hepatitis made up 72.5% of all registered contagious and invasive diseases: HBV was diagnosed in 46%, HCV in 50.2% and HBV + HCV in 1.8% of all viral hepatitis cases. Dermatoses, mostly of allergic etiology, were the second most prevalent diseases (11.4%), and were most frequently associated with exposure to latex, thiurams, mercaptobenzothiazole and non-specified rubber compounds-73% of all factors causing allergic dermatoses. Chronic diseases of locomotor system, chronic diseases of peripheral nervous system, chronic diseases of bronchi, chronic inflammation of nose, pharynx, larynx and trachea, and intoxications were also reported. Almost twofold decrease in the incidence rate in the population of workers referred to "Health and social work" activity section was observed in 2001 compared to 1994. The decrease in the number of the registered occupational diseases seen in the hospital employees was mostly due to the effective anti HBV prevention programs carried out in Poland among health care personnel since 1989. PMID- 12577804 TI - [The incidence of asbestosis in Poland]. AB - The aim of the work was to analyze the incidence of occupational asbestosis. The analysis covered all cases of asbestosis reported to the Central Register of Occupational Diseases during the years 1970-2001. The cases were classified by patients' gender, age, place of employment, duration and magnitude of exposure to asbestos dust. In all, 1779 cases of asbestosis were registered, including 1200 male cases. During the period under study, a growing number of diagnosed cases were observed (from several cases in the early 1970s to several dozens in the 1990s). During the period of 32 years, the average incidence was 0.5 cases per 100,000 employees, and asbestosis contributed in 5.4% to all occupational pneumoconioses. The majority of asbestosis cases was diagnosed in workers employed in the plants of asbestos processing (38.3%) and asbestos-cement products, followed by foundries (13.1%) and shipyards (10.5) workers. The mean age of asbestosis patients was 55 years, and the average duration of employment under exposure to asbestos was about 21 years. The data recorded in the certificates of occupational disease show that asbestos dust concentrations at workplaces ranged from 1.4 to 38.3 mg/m3 (median, 3.8 mg/m3). Shipyard workers were exposed to the highest (Me = 8.0 mg/m3) and asbestos-cement workers to the lowest (Me = 2.8 mg/m3) concentrations. PMID- 12577806 TI - [Occupational hearing loss: new principles of certification]. AB - Noise-induced hearing loss, together with presbyacusis, are the most prevalent causes of the functional impairment of the inner ear in an adult population. The authors discuss briefly the epidemiological, diagnostic and medical certification issues. The paper is focused on the new legislation that brings about considerable changes in the procedure of medical certification of noise-induced hearing loss. Both the definition of the pathology and new principles for its certification as an occupational disease were intended to harmonize Polish regulations with respective legislation of the EU countries. PMID- 12577805 TI - [Exposure of patients as a criterion justifying radioisotope examinations (in vivo)]. AB - Exposure of patients, known as medical exposure, is an inevitable consequence of the diagnostic and therapeutic application of ionizing radiation. This includes among others radioisotope examination in vivo. The authors evaluate diagnostic radioisotope examinations in vivo, as an artificial source of patients' exposure, in comparison with radiological diagnostics, the most common type of medical exposure, and diagnostic methods, employing physical agents other than ionizing radiation (ultrasound, magnetic field). It is expected that the results of the evaluation will provide the justifying grounds for a sensible choice of a diagnostic method among those available. On the basis of the precise data collected in 21 laboratories, the frequency of radioisotope examinations during the years 1998-2000 was estimated at about three per 1000 inhabitants. It was found that the examination of the thyroid gland, skeleton and heart were performed most frequently. The information gathered and the literature data helped to calculate an average dose per one inhabitant of Poland. The values are as follows: average effective dose per one examination--6 mSv; average effective dose per one inhabitant--0.033 mSv. It should be stressed that apart from the examinations of the thyroid gland performed with use of 1311 (16% of the total number of examinations), the average effective dose received by patients during other examinations is equal to only 2.2 mSv. Finally, the authors present the data on patients' exposure during radioisotope examinations and diagnostic examinations employing other methods. They indicate that many radioisotope examinations provide valuable diagnostic information that could not be obtained by means of other methods. Moreover, these examinations involve relatively low levels of effective doses received by patients. PMID- 12577807 TI - [Assessment and analysis of the acoustic environment of soldiers exposed to impulse noise]. AB - The aim of the study was to assess the acoustic environment of soldiers attending a one year basic military service. The study material comprised 13 types of weapons used by this group of soldiers. During the target practice, the following parameters were measured separately for the right ear and the left ear: equivalent sound pressure level A (LA eq, Te); maximum sound pressure level A (LA F max) and peak sound pressure level C (LC peak). The measurements covered several single shots or a shot series. In addition, a spectroanalysis in 1/3 octave band frequency of 40-50,000 Hz, was conducted. There were following values of measurements in the direct vicinity of the ears: LA eq, Te fell within the range of 106.2-119.5 dB (mean, 112.2 dB) for the right ear and 104.2 dB-118.4 dB (mean, 113.2 dB) for the left ear; LA F max within the range of 124.5-132.3 dB (mean, 128.3 dB) for the right ear and 116.3-135.1 dB (mean, 128.1 dB) for the left ear; LC pcak within the range of 151.8-156.5 dB (mean, 154.8 dB/155 dB) for the right ear and 151.9-156.2 dB (mean, 155.4 dB) for the left ear. In the noise spectrum, components of audible frequencies predominated, 160-300 Hz (mainly for high caliber weapon); 1600-6300 Hz for the right ear and 2500-4000 Hz for the left peak (small caliber weapons); 6300-16,000 and ultrasonic 25,000-50,000 Hz. No significant differences were found in the measurements of the right ear and the left ear. PMID- 12577808 TI - [Effects of individual features on the measurement of vibration perception thresholds: standard setting for healthy people]. AB - The thresholds of vibration perception in healthy people may differ significantly depending on individual and constitutional features: age, weight, height, race and gender, and also on various addictions like smoking or alcohol abuse. These variables have not as yet been analyzed in setting Polish standards of vibration perception thresholds used for therapeutical and certification purposes. The aim of the study was to develop a model that could render it possible to assess the normative values of vibration perception, taking account of individual features. The study covered 187 healthy persons free from exposure to vibration. Two methods were used to determine vibration perception thresholds: the standard Polish method and the method based on the ISO 13091-1/2001 standard. The methods differed in the technical parameters (contact force of vibrating probe 1.2 N and 0.1; probe diameter 12 and 5 mm-standard method and the method according to ISO, respectively), the presentation of stimuli (ascending method versus von: Bekesy method) and their frequencies. Vibration perception thresholds were significantly influenced by age (within the range of 63-250 Hz-standard method; 4-250 Hz-method according to ISO), body mass (full range of frequencies--standard method; 4-125 method according to ISO) and height (single frequencies--both methods). A model for determining vibration perception thresholds, taking account of age, height and body mass of study subjects, was developed. The results of the study show that individual and constitutional features should be taken into consideration when interpreting the results of the vibration perception examinations conducted for the purpose of occupational disease certification. PMID- 12577809 TI - [Dust exposure in a pottery plant assessed by using GRIMM dust monitor]. AB - In one of the pottery plants in which pneumoconiosis was the only certified occupational disease (16 cases during the years 1984-1997), the concentrations of dust in inhalable, thoracic and respirable fractions were measured at most characteristic workplaces. The highest concentrations in all the three fractions was found in the slip house; the mean concentrations were as follows: inhalable dust-7231.8 mg/m3; thoracic fraction--4834.9 mg/m3; and respirable fraction 1402.2 mg/m3. High concentrations of all the dust fractions were also observed at the workplace of the intershop transport worker who removed the waste body from cast and throw shops. The largest number of pneumoconiosis (7) cases was found among the workers of the slip house (20 persons employed); this was followed by the intershop transport workers-4 cases (4 persons employed). At the other workplaces (cast shop, throw shop and firing), concentrations of inhalable dust ranged from 1175 to 3232 mg/m3; thoracic from 726 to 2001 mg/m3; and respirable from 146 to 549 mg/m3. The contents of free crystalline silica (SiO2) in the air dust ranged from 3% in firing to 50% in slip house, in the latter mostly in the form of quartz or cristobalite. The main fraction of inhalable dust was the tracheobronchial fraction, ranging from 43.1 to 48.2%, the extrathoracic fraction ranged from 32.8 to 42.3% and respirable fraction from 11.8 to 21.6%. On the basis of the graphic record, such activities as pouring of raw materials into grinding mill and reloading of the waste body were identified at the workplaces of slip making and internship transport as those associated with the highest dust emission. PMID- 12577810 TI - [Diagnosis and certification of the ability of epileptic patients to drive motor vehicles: cases consulted by the author]. AB - Epilepsy is a frequent diagnostic problem. It is also difficult to certify whether an epileptic patient is able to drive a motor vehicle. With the advent of efficient anti-epileptic treatment, a general practice of refusing epileptic patients driving license should be seriously reconsidered. However, the matter should be given careful consideration not to jeopardize public safety and patients' rights. The aim of the study was to highlight the problems encountered in rediagnosing and certifying people with diagnosed epilepsy or pseudoepileptic seizures. The authors discuss the diagnosis and certification procedures in persons with epileptic seizures after severe craniocerebral trauma. They also analyze a case of diagnosed epilepsy suggesting the syncope in a patient with cardiac defect; a case of psychogenous pseudoepileptic seizures and the course of the disease in a patient with febrile convulsions in childhood. The problems result from the fact that reliable medical histories are not available and thus the retroassessment of the clinical picture of epileptic seizures is not possible. Missing results of timely laboratory tests (EEG, ECG) and diagnostic errors concerning earlier episodes, especially epilepsy diagnosed inconsiderately, are additional obstacles. PMID- 12577811 TI - [Occupational and non-occupational aspects of carpal tunnel syndrome]. AB - Carpal tunnel syndrome is caused by the pressure of the medianus nerve in the carpal canal. This syndrome is characterized by the following symptoms: numbness of the thumb and fingers, hand pain, particularly at night, and handicap of the hand efficiency. The examination of the perception, provocation tests, electromyography, carpal radiography, ultrasonography and magnetic resonance imaging are very helpful in the diagnosis of carpal tunnel syndrome. Pathological changes in flexor's theca are the most frequent causes of the syndrome. It may also occur in patients with lupus erythematosus, tuberculosis, endocrinopathy, rheumatic diseases and during pregnancy. Carpal tunnel syndrome is one of the most common occupational neuropathy. It is associated with the recurrence of carpal movements and observed in seamstresses, butchers, persons testing bar codes in supermarkets and meat packers. PMID- 12577812 TI - [Prevention of hand-arm vibration syndrome by using antivibration gloves]. AB - Hand-arm vibration syndrome induced by the exposure to mechanical vibration transmitted from power tools to the operator's hands contributes highly to the number of occupational diseases certified in Poland, as well as in other countries. Use of antivibration gloves is one of the methods for reducing the occupational risk of vibration damages. However, the number of researchers express certain reservations about the benefits from wearing such a protective device, mostly because of very limited scientific and explicit evidence to prove the effectiveness of antivibration gloves in preventing hand-arm vibration syndrome. But there is no controversy about the opinions that to achieve a maximum degree of protection, antivibration gloves should be selected to match particular vibratory tools, taking account of the vibration spectrum generated, by them. The paper presents the general principles of the glove selection, the results of the studies of the effectiveness of antivibration gloves carried out by the Central Institute for Labour Protection (CIOP), as well as some other results, opinions and ideas. The gathered data prove that antivibration gloves are a useful measure to prevent hand-arm vibration syndrome. PMID- 12577813 TI - [Polish bibliography of occupational medicine before 1950]. AB - The paper gives the data on the first Polish journal devoted to issues concerning occupational medicine: Editorial Board, Publishers, contents of the first issues, and a presentation of the papers published. The authors invites the Editorial Office to open in the Journal a new section on the history of industrial medicine. PMID- 12577814 TI - [The VIII International Conference "Neurobehavioral Methods: Influence on Occupational and Environmental Health" (Feb. 23-26, 2002, Brescia, Italy)]. PMID- 12577815 TI - Quality-based incentives may change capitation landscape. PMID- 12577816 TI - Medicare rates confirm New England group's success. PMID- 12577817 TI - Study demographics, practice patterns to predict office-based service use. PMID- 12577818 TI - Top 100 hospitals continue to raise the bar on quality, costs. PMID- 12577819 TI - Cry freedom. PMID- 12577820 TI - Nothing to it. PMID- 12577822 TI - Delayed discharges. Another fine mess. AB - A simulation exercise on charging for delayed discharges revealed significant potential for conflict between organisations, and no benefit for service users. Participants thought the charging system would work against the development of community services. They favoured joint NHS/local authority approaches to the development of an effective system for preventing delayed discharges. PMID- 12577823 TI - Major incidents. Norwalk on the wild side. AB - An outbreak of the Norwalk-like virus at an acute hospital in October affected 147 patients and 200 staff. It was brought under control within two weeks, following the establishment of a team to manage the incident. Some day surgery was cancelled, but no inpatient operations were affected. The public was discouraged from visiting the hospital during the outbreak. Local GPs were co operative about managing patients with the virus at home. Some doctors came to work with symptoms, despite the trust urging affected staff to stay at home. PMID- 12577824 TI - Primary care trusts. The human touch. AB - Research with six primary care trusts, started in 2001, suggests that many lack access to relevant expertise in human resources management. Where PCTs do have human resources directors, they are often being asked to act on strategic issues which are outside their experience. PCTs required to rely on acute trusts' HR departments for HR expertise find this unsatisfactory as the issues facing the two sectors are so different. The status of HR management in PCTs needs to be boosted. PMID- 12577825 TI - Buildings. Fast forward. PMID- 12577826 TI - Buildings: sustainability. Atmospheric pressure. PMID- 12577827 TI - Buildings: sustainability. Driver for change. PMID- 12577828 TI - [Insulin and arterial hypertension: role of hypersympathicotonia]. PMID- 12577829 TI - [Study of mortality due to myocardial infarction and alcoholism on the basis of the WHO MONICA and Acute Myocardial Infarction Registry programs]. AB - AIM: To study trends of mortality due to alcoholism (A) and cardiovascular diseases (CVD) in Novosibirsk in 1981 to 1998. MATERIALS AND METHODS: The WHO MONICA and Acute Myocardial Infarction Register programmes were used to survey the population of 3 Novosibirsk districts. There were notified 9016 cases of nonviolent death, by exclusing another pathology, with the exception of CVD and alcoholic intoxication. RESULTS: Mortality rates due to myocardial infarction (MI) were relatively stable throughout the observation period, except for 1988, 1994, and 1998 (a significant increase). Those due to A showed the following trend: stabilization in 1981-1982, a significant increase in 1983-1987, stabilization in 1988-1991, a significant increase in 1992-1994, and a decrease in 1995-1998. Mortality rate from MI were 2-3 times greater than that from A, with the exception of 1994-1995 when they were equal, i.e. the trends of mortality from MI and A did not coincide. Social stress is a factor that greatly influences MI death rates, mainly due to prehospital mortality. As social stress increases, younger age groups, both males and females, are afflicted, which is extremely hazardous for the population. By taking into account the mortality trends, it may be stated that A hardly affects death rates from MI. In the pattern of mortality from CVD, death rates from MI and A were 50-70% in males and 35-60% and females; those from MI were 35-55 and 30-45%, respectively. The increase in alcoholism mortality is associated with the population's addictive behavior in the period of social upheavals in the community. CONCLUSION: According to official statistics, the mortality trends do not reflect the actual state of things. This may be done only by stringently standardized programmes with their schemes of data collection and diagnosis verification. The WHO MONICA and Acute Myocardial Infarction programmes belong to such programmes. PMID- 12577830 TI - [State of lipid peroxidation, blood antioxidant defense in patients with myocardial infarction aggravated by circulatory insufficiency]. AB - AIM: To study free radical processes in patients with myocardial infarction (MI) aggravated by heart failure. MATERIAL AND METHODS: Forty seven patients taken to the clinic within the first 5 hours of MI were examined. The patients were divided into 2 groups: 1) those with left ventricular failure-complicated IM (n = 25); 2) those with uncomplicated MI (n = 22). A control group included 17 apparently healthy males. The activity of lipid peroxidation (LPO) and blood antioxidative defense was determined in the patients' red blood cells prewashed off the plasma with saline solution at a temperature of 4 degrees C. RESULTS: The tension of free radical lipid oxidation mechanisms in patients with MI aggravated by circulatory insufficiency is followed by an increased antiradical activity. In patients with uncomplicated higher LPO activity is attended the less marked activation of antioxidative enzymes. The examinees were found to show a correlation of the global systolic function of the left ventricle with the concentration of LPO products and with the activity of antioxidative enzymes. CONCLUSION: More significant LP activation coupled with left ventricular systolic dysfunction was noted in patients with MI aggravated by acute heart failure. PMID- 12577831 TI - [Role of left ventricular systolic and diastolic dysfunction in clinical manifestations of chronic heart failure in patients with prior myocardial infarction]. AB - AIM: To study the impact of systolic and systolic dysfunction of the left ventricle (LV) on the clinical manifestations of chronic heart failure (HF) in patients with prior myocardial infarction (MI). MATERIALS AND METHODS: 102 patients (mean age 47.4 +/- 0.7 years) who had sustained large focal IM at least 6 months before were examined. LV echocardiography was made to define systolic diastolic relationships in HF. RESULTS: Analyzing the prevalence of primary diastolic HF in patients with its different clinical manifestations, it may be concluded that LV diastolic dysfunction is of the leading pathogenetic value at early stages of ischemic heart dysfunction. A complex relationship between the clinical manifestations of HF and a number of parameters characterizing the systolic and diastolic function of LV was estimated by the multiple regression analysis and presented as a trinomial equation. CONCLUSION: The results of the regression analysis have shown that the clinical manifestation of HF intensifies as relaxation slows down and transmitral blood flow redistributes to the left atrial systole; the severity of HF also increases with the depletion of compensatory mechanisms responsible for LV contractility. PMID- 12577832 TI - [Relationship of the duration of QT interval and corrected QT interval in patients with coronary heart disease and the cause of the bicycle ergometer test cessation]. AB - AIM: To comparatively study Q-T interval duration in groups of patients with coronary heart disease (CHD) identified by the cause of exercise test cessation. MATERIALS AND METHODS: Bicycle ergometric test (BET) was performed in 93 males with CHD (functional class II-III angina pectoris on exertion). During BET, ECG recorded in the leads, as described by Nebu. The QT interval was visually measured in one lead. This interval was determined as a distance between the onset of a Q wave and the end of a T wave (in mlsec). The mean value by 3 cardiac cycles was taken as an actual value of the QT interval. To compare the duration of the QT interval at different rates of cardiac rhythm, its corrected value (QTc) was calculated. RESULTS AND CONCLUSION: In case of positive BET (anginal attack, 1-mm or more depressed ST segment at a distance of 0.8 cm from the joint J), the values of QT and QTc measured at rest and peak exercise were greater than those if the cause of test cessation was the submaximum heart rate or a patient's fatigue. The value of QTc was higher if the cause of BRT cessation was 1-mm or more depressed ST segment preceding an anginal attack or the only cause of test cessation. PMID- 12577833 TI - [Pulse blood pressure (according to 24-hour monitoring) and left ventricular myocardial structural changes in patients with hypertensive disease]. AB - AIM: To study a relationship of the magnitude of structural changes in the left ventricle (LV) to the mean daily pulse blood pressure (PBP) in patients with hypertensive disease (HD). MATERIALS AND METHODS: 70 male patients (mean age 49 +/- 1 years) with stages I (n = 54) to 11 (n = 16) HD. LV mass (LVM) was estimated by echocardiography according to the formula derived by R. B. Devereux et al. and normalized to body surface area [the LVM index (LVMI)]. The relative thickness index (RTI) for the posterior wall (PWRTI) and ventricular septum (VSRTI) was calculated as a ratio of the sum of PWRTI and VSRTI to the LV end diastolic size. LVMI > 125 g/m2 was considered to be a criterion for LV hypertrophy (LVH). 24-hour blood pressure (BP) monitoring was performed with a Spacelabs-90207 device (USA). According to the 24-hour PBP value, the patients were divided into 2 groups: 1) those (n = 17) having PBP24 > 53 mm HG and 2) those (n = 53) having PBP24 < 53 mm Hg. RESULTS: Group 1 patients were found to have significantly higher LVMI, LV WRTI, and incidence of LVH and a complex of changes in the BP profile as higher values of 24-hour systolic, diastolic and mean BP, PBP, and BP variations. Multiple regression analysis revealed a highly significant contribution of PBP24 to the development of LVH. CONCLUSION: The pedictive value of PBP as an index that characterizes a dynamic pressure load in regard to LV structural changes is higher than that of mean BP as a static load index and a BP variation index. PMID- 12577834 TI - [Tissue insulin resistance and endothelial function of large vessels in patients with arterial hypertension]. AB - AIM: To examine insulin resistance and hyperinsulinemia in obese patients with moderate arterial hypertension (AH) without diabetes mellitus, glucose intolerance, and ischemic heart disease; to evaluate the endothelial vasomotor function of large vessels. MATERIAL AND METHODS: 41 males (mean age 49.3 +/- 5.4 years) with AH were examined by anthropometry, measurements of the levels of glucose, immunoreactive insulin, C-peptide, and biochemical blood tests. Endothelial vasomotor function was assessed ultrasonically. RESULTS: Insulin resistance preceding abnormal glucose tolerance was found in 52.4% of the patients with AH and obesity. In these patients lipid metabolic disturbances were more pronounced than in those without hyperinsulinemia. There was endothelial vasomotor dysfunction with preserved capacity of arteriolar and capillary walls for vasodilatation irrespective of the presence or absence of insulin resistance. CONCLUSION: Insulin resistance and endothelial dysfunction are most common in hypertensive patients with obesity. PMID- 12577835 TI - [External respiratory function in stages I and II arterial hypertension]. AB - AIM: To examine external respiratory function (ERF) and pulmonary circulatory hemodynamics in patients with Stages I and II arterial hypertension (AH). MATERIALS AND METHODS: 105 patients with AH were examined. In 43 of them, spirography and echocardiography were performed with pulmonary pressure and determined during elevated blood pressure (BP) and antihypertensive therapy. RESULTS: ERF disorders that recovered with BP lowering were detected in 62.8% of patients with AH. There were also signs of elevated pulmonary pressure. CONCLUSION: In patients with HF, pulmonary hemodynamic disorders, changes in ERF, and the degree of AH are closely interrelated. PMID- 12577836 TI - [Possibilities of achieving target blood pressure in patients with essential hypertension in relation to renal functional status and age]. AB - AIM: To study renal functional changes in patients with essential hypertension (EH) and normal blood creatinine levels during antihypertensive therapy with currently available drugs to achieve blood pressure lower than 140/90 mm Hg. MATERIALS AND METHODS: Renal functional changes were studied in 114 patients with EH and normal blood creatinine levels during antihypertensive therapy aimed at achieving blood pressure lower than 140/90 mm Hg. Glomerular filtration rate (GFR) was calculated by the Cockcroft-Gault formula and blood creatinine levels were measured before and 6, 12, 24, 52 weeks of treatment. RESULTS: Renal dysfunction was detected in 43.9% of the patients as GFR < 80 ml/min in 35.1% and hyperfiltration (GFR > 140 ml/min) in 8.8%. The patients with baseline decreased GFR were older, had a longer history of EH, and lower body-mass index than those with hyperfiltration (p < 0.05). There was a considerable renal functional improvement in patients with baseline decreased GFR and achieved BP < 140/90 mm Hg. Target BP could be achieved only in 20% of the patients with hyperfiltration. Antihypertensive therapy caused reduced hyperfiltration in these patients. In patients aged over 60 years who had a baseline GFR < 80 ml/min, achievement of target BP also promoted renal functional improvement. CONCLUSION: Most patients with EH and normal blood creatinine levels have renal dysfunction. Normalization of BP substantially improves renal functional parameters irrespective of age. PMID- 12577837 TI - [Functional and biochemical characteristics of the vasodilator effects of nebivolol in patients with arterial hypertension]. AB - AIM: To examine nebivolol-induced changes in endothelial vasomotor function and blood concentrations of nitric oxide (NO) metabolites in patients with arterial hypertension (AH). MATERIALS AND METHODS: The impact of 8-week nebivolol monotherapy on 24-hour blood pressure (BP) monitoring, ECG, autonomic heart regulation, changes in endothelium-dependent vasodilation (EDVD) and endothelium independent vasodilation, on the plasma concentration of NO metabolites, and on the endothelial responsiveness index (ERI) were studied in 30 patients with stages I-II AH. RESULTS: Nebivolol provides a sufficient 24-hour BP monitoring, reduced heart rate, the incidence of supraventricular arrhythmias, and left ventricular myocardial mass with the optimization of autonomic cardiac regulation. The agent recovers NO-ergic regulation of endothelial vasomotor function in patients with AH by normalizing the basal level of NO and EDVD. The study of plasma NO metabolites before and during reactive hyperemia test with ERI calculation substantially extends an idea of the mechanisms of EDVD and supplements the diagnostic potentialities of this method. CONCLUSION: Dysfunction of the vascular endothelial NO-producing system occurs in patients with AH. In addition to its positive effect on 24-hour BP profile and myocardial morphofunctional parameters, nebivolol restores the NO-ergic regulation of endothelial vasomotor function in patients with AH, by potentiating NO expression, and modulates steady-state vasodilation. PMID- 12577838 TI - [Tonocardin in complex treatment of diabetes mellitus concurrent with hypertension]. AB - AIM: To examine the effect of the alpha 1-adrenoblocker tonocardin (doxazosin) on the course of arterial hypertension (AH) and on carbohydrate and lipid metabolism and insulin resistance in patients with type 2 diabetes mellitus (DM) concurrent with AH. MATERIALS AND METHODS: 18 patients with type 2 DM concurrent with AH, treated with tonocardin for at least 12 weeks were examined; the fasting glycemia, the levels of blood glycosylated hemoglobin, serum total cholesterol and triglycerides, and the degree of insulin resistance (intravenous insulin load or insulin tolerance test and the insulin resistance index estimated by the HOMA method) were determined. RESULTS: Tonocardin treatment lowered systolic BP (from 159 +/- 19.83 to 136.57 +/- 17.43 mm Hg; by 14.5%), diastolic BP (from 93.38 +/- 12.98 to 79.12 +/- 11.69 mm Hg; by 15.28%), fasting glycemia (from 9.32 +/- 1.61 to 7.05 +/- 1.51 mmole/l; by 24.36%), glycosylated hemoglobin Ai (from 9.63 +/- 1.86 to 8.59 +/- 0.98%; by 10.8%), total cholesterol (from 6.09 +/- 0.57 to 5.4 +/- 0.4 mmole/l; by 11.4%), triglycerides (from 2.11 +/- 0.57 to 1.88 +/- 0.52 mmole/l; by 11%), glycemia after 30-min insulin load (from 9.32 +/- 1.61 to 5.77 +/- 1.57; by 39% and from 7.05 +/- 1.51 to 4.2 +/- 1.25 mmole/l; by 44% at the beginning and end of the follow-up, respectively), insulin resistance index (from 9.87 +/- 2.45 to 6.57 +/- 1.99; by 33.5%). CONCLUSION: The findings suggest that tonocardin exerts an antihypertensive effect and positively affects carbohydrate and lipid metabolisms, and diminishes insulin resistance in patients with type 2 DM concurrent with AH. PMID- 12577839 TI - [Use of plasmapheresis in the treatment of drug-resistant cardiac arrhythmias]. AB - AIM: To study the therapeutical efficiency of plasmapheresis (PA) in patients with drug-resistant cardiac arrhythmias (CA) and its mechanisms. MATERIALS AND METHODS: Discrete PA sessions were carried out in 56 patients with drug-resistant CA: paroxysmal atrial arrhythmia (AA), ventricular and supraventricular premature contractions, supraventricular tachycardia of various etiology. Biochemical blood values, coagulographic parameters, lipid peroxidation (LPO), the spectrum of nonesterified fatty acids (NEFA), the level of medium-sized molecules were determined, ECG monitoring, EchoCG, and left ventricular radioisotope computed tomography were performed before and after a PA session. RESULTS: PA was effective in 50% of cases. The duration of its effect averaged 3.0 (1.25-5.0) months. PA was more beneficial for patients with IHD, AA with normal left atrial dimensions, and hyperlipidemia. The duration of the effect was significantly higher when antiarrhythmic drug therapy was continued after PA. Due to PA, there were significant decreases in the blood concentrations of cholesterol, medium sized molecules, malonic dialdehyde (MDA) and in the proportion of polyunsaturated NEFA. The antiarrhythmic effect was associated with the decreases in MDA and NEFA, with a tendency for a reduction in the rate of chemiluminescence. CONCLUSION: PA may be used in the treatment of drug-resistant CA. The most significant mechanism of its antiarrhythmic activity is to recover sensitivity to antiarrhythmics. The intrinsic antiarrhythmic activity may be associated with its effect on NEFA metabolism and LPO; however, its mechanisms await further studies. PMID- 12577840 TI - [A patient with mitral prolapse: differential intervention programs in general medical practice]. AB - AIM: To develop and implement intervention programmes for patients with mitral prolapse (MP) under real conditions of general medical practices of Russia in the transition period. MATERIAL AND METHODS: The MP register covered 208 patients among the adult population (n = 3346) of 2 districts served by general practitioners. According to the formalized protocols, clinical, instrumental, and psychological studies were conducted, which included examination of complaints and physical status, electrocardiography, echocardiography, paired echocardiography before and after bicycle ergometry exercise tests, psychological testing, study of the social status and life quality of patients. RESULTS: According to clinical and instrumental findings, the patients were divided into 4 groups, where differential individually based intervention programmes were implemented; the latter included modules of family upbringing, family prophylaxis, drug and nondrug treatment. Psychotherapeutical potentiation was performed in patients with the low cooperation index on an individual basis in accordance with revealed personality traits. Clinical symptoms diminished in severity, scores on pathological disease attitude scales lowered, life quality increased, persons with the low cooperation index reduced in number. CONCLUSION: The developed and tested intervention programmes in patients with MP are effective and may be successfully implemented in general medical practice facilities. PMID- 12577841 TI - [Lipoprotein(a) and Apo-A isoforms in patients with intermittent claudication]. AB - AIM: To study a correlation between lipoprotein(a) [LP(a)], as well as the phenotype of apoprotein(a) [apo(a)] and the presence of intermittent claudication (IC). MATERIALS AND METHODS: The study included 25 patients (9 females, 16 males; mean age, 63.4 +/- 8.2 years) and 50 individuals (21 females, 29 males; mean age, 61.3 +/- 6.1 years) of a control group. RESULTS: In the patients, the level of LP(a) was 3 times higher than in the controls (median 39 and 13 mg/dl, respectively; p < 0.01). The low molecular-weight phenotypes of apo(a) occurred significantly more frequently in the patients than those in the controls (62 and 28%, respectively; p = 0.02). CONCLUSION: Multifactorial analysis has shown that the level of Lp(a) is an independent and important factor associated with the presence of IC. PMID- 12577842 TI - [Changes in the cardiac valvular apparatus in patients with end-stage chronic renal failure on long-term programmed hemodialysis]. AB - AIM: To examine the specific features of development of cardiac valvular diseases, biochemical changes in the blood composition, and changes in the density of bone tissue in patients with chronic renal failure (CRF) treated with programmed hemodialysis. MATERIALS AND METHODS: 118 patients with CRF (Group 1 patients (n = 69) aged 18-40 years and Group 2 patients (n = 49) aged 55-73 years) were examined. RESULTS: Mitral valvular prolapse or disease mainly developed in Group 1 patients having profound phophorous and calcium metabolic changes and significantly elevated concentrations of blood parathyroid hormone; developed aortic valvular disease and calcium salt-impregnated cardiac tissues were observed in Group 2 patients. CONCLUSION: The revealed regularities occur in chronic renal failure regulated by programmed hemodialysis and they are risk factors for chronic heart failure in elderly patients in particular. PMID- 12577844 TI - [Specific features of intracardiac relationships according to echocardiographic data in patients with obstructive or restrictive ventilation disorders]. AB - AIM: To examine relationships between the areas portions and hemodynamic parameters according to echocardiographic data in patients with various lung diseases accompanied by obstructive and restrictive ventilation disorders. MATERIALS AND METHODS: 154 patients aged 15 to 67 years who had various lung diseases were examined. Expiratory respiratory function and echocardiography were performed in all the patients. RESULTS: Despite the type of ventilation disorders, all cardiac areas are closely interrelated. The specific features of these relationships depending on the type and severity of ventilation disorders are revealed. Thus, the correlations of the right ventricle with other cardiac areas in the obstructive type increase with the greater severity of bronchial obstruction and they are determined by the diastolic size (volume) of this ventricle. CONCLUSION: The findings suggest that there are two types of compensatory reactions of the heart in lung diseases with obstructive and restrictive ventilation disorders. PMID- 12577843 TI - [Physiological analysis of plasma hypo-osmolality in pneumonia]. AB - AIM: To study the physiological mechanisms of plasma hypoosmolality in patients with pneumonia and on this basis to elaborate principles of therapy for this condition. MATERIALS AND METHODS: 52 individuals of different age, including 26 patients with pneumonia, were examined. Osmolality, the concentrations of ions of sodium, potassium, magnesium, and creatinine were measured in the serum. RESULTS: The patients with pneumonia were found to have osmolality, hyponatremia in combination with severe hypodiuresis, high urinary osmotic pressure and intensive reabsorption of osmotically free water in the kidney, which leads to blood dilution. As hypoosmolality usually causes higher diuresis and decreased urinary osmolality; hypodiuresis with high urinary osmolality in pneumonia is indicative of effective renal performance and its altered regulation evidently due to the hypersecretion of vasopressin or to the decreased formation of a number of autacoids in the kidney. CONCLUSION: Blood hypoosmolality and hyponatremia in the examined patients result from inadequate blood osmolality and high urinary osmotic concentrating. The principles of this condition in pneumonia are discussed and aquaretics are proposed for use as pathogenetic therapy. PMID- 12577845 TI - [Hemodynamic features in bronchial asthma concurrent with hypertensive disease]. AB - AIM: To study the cardiorespiratory system, central and intracardiac hemodynamics. MATERIALS AND METHODS: 274 patients with bronchial asthma (BA), including 117 patients with this condition concurrent with hypertensive disease (HD) during traditional medical treatment were examined. RESULTS: The impact of concomitant HD on the pattern of performance of the cardiovascular system and the development of the cardiac structure in these patients was evaluated. CONCLUSION: In patient with BA concurrent with HD, external respiratory dysfunction diastolic dysfunction of both ventricle deteriorate, specific peripheral resistance enhances. PMID- 12577846 TI - [Effects of small-dose cytarabine on the course of atheromatosis in over 60 years of age patients with acute myeloblastic leukemia (description of 2 cases)]. PMID- 12577847 TI - [The cardiovascular system of patients with lymphogranulomatosis in long-time remission]. AB - AIM: To study the cardiovascular system in patients with lymphogranulomatosis (LGM) at prolonged remission. MATERIALS AND MATERIALS: 78 patients received radiation and chemotherapy for lymphogranulomatosis in 1973 to 1995. All the patients underwent electro- and echocardiography (EchoCG), 40 patients had radioisotopic ventriculography (RVG). RESULTS: EchoCG revealed nonspecific changes in the ST segment in 19 (24.3%) patients. Systolic function was decreased vto 46% only in 1 patient. RVG showed a significant reduction in ejection fraction in the experimental group as compared to the control one (55.5 +/- 9.5 and 68.4 +/- 8.0, respectively; p < 0.05). CONCLUSION: Cardiac disorders revealed in patients with LGM at prolonged remission are mainly insignificant. The risk for cardiac events is not a guide for reducing radiation doses and the intensity of chemotherapy. PMID- 12577849 TI - [Chronic and lingering Yersinia ileitis]. AB - AIM: To study the incidence, clinical manifestations, and diagnosis of chronic and lingering Yersinia ileitis. MATERIALS AND METHODS: 82 patients with pains in the right iliac area. The coagglutination test was used to reveal Y. pseudotuberculosis and Y. enterocolitica antigens as part of circulating immune complexes in the serum and coprofiltrates. RESULTS: Correct diagnosis was established 3 months to 2 years after the acute period of the disease. After etiotropic treatment, 2 patients continued to have the steady pain syndrome in the right iliac area, which is likely to be associated with periileitis and, 46 patients developed chronic Yersinia ileitis, as evidenced by their positive serological reactions to antigens and iliac morphological findings. Development of a residual phase may be presumed in 2 patients who continued to have stool disorders and abdominal pain, but no pseudotuberculous antigens in the stool and blood. CONCLUSION: Patients with lingering (up to 3 months) and chronic (over 3 months) Yersinia ileitis were 38.1% of the total number of those with ileocecal pain. The chronic nature of Yersinia ileitis is associated with late diagnosis, developed periileitis, and immunity disorders. PMID- 12577848 TI - [Time course changes in the count of peripheral granulocytes in children with acquired aplastic anemias treated with antithymocyte globulin, cyclosporin A and granulocytic colony stimulating factor]. AB - AIM: To examine the pattern of changes in the count of peripheral granulocytes in children with aplastic anemias (AA), receiving a combined immunosuppressive therapy with antithymocytic globulin (ATG) and cyclosporin A in combination with granulocytic colony-stimulating factor (G-CSF). MATERIALS AND METHODS: 31 children (17 boys and 14 girls) aged 2-15 years (median 9 years) with newly diagnosed severe and very severe acquired AA took a combined immunosuppressive therapy with ATG and cyclosporin A in combination with G-CSF in an initial dose of 10 micrograms/kg a day. RESULTS: A three-linear and response was recorded in 19 (61%) children, an isolated granulocytic response was in 26 (84%). The interval median before the recovery of granulocytes to 1.5 x 10(9)/l and 5 x 10(9)/l was 19 and 38 days, respectively. CONCLUSION: Use of G-CSF may increase the count of granulocytes in the vast majority of patients with AA, without dramatic influence on the frequency of a three-linear response. Intermittent use of G-CSF may maintain the count of granulocytes long at the safe level and reduce the cost of treatment. PMID- 12577850 TI - [Serum lipid spectrum in HBsAg carrying patients with alcoholism]. AB - AIM: To reveal the effect of HBsAg on the serum lipid spectrum in alcoholics. MATERIALS AND METHODS: The serum lipid spectrum was studied in 30 alcoholics who were HBsAg carries, 50 alcoholics without HBsAg and 40 healthy individuals. RESULTS: Unlike alcoholic patients without HBsAg, those who were HBsAg carriers had higher relative content of phospholipids, lysophosphatidylserine, phosphatidylcholine, phosphatidylethanolamine and lower levels of cholesterol ester, lysophosphatidylcholine, lysophosphatidylethanolamine, sphyngomyeline. CONCLUSION: HBsAg exerts an additional unfavorable effect on the serum lipid spectrum in patients with alcoholism. PMID- 12577851 TI - [Radiation-modifying effect of oxygen barotherapy in complex treatment of patients with rheumatoid arthritis and osteoarthrosis]. AB - AIM: To evaluate the efficiency of radiotherapy (RT) in combination with oxygen barotherapy (OBT) versus PT used in the complex therapy for rheumatoid arthritis (RA) and osteoarthrosis (OA). MATERIALS AND METHODS: 46 patients with RA and 18 patients with OA were examined, of them 24 patients with RA and 10 with OA received complex therapy involving a course of OBT. The groups of patients had no statistically significant differences in the characteristics of their disease and in the nature of the therapy performed. All the patients received RT as radiation therapy. The patients were followed up for 2 years by assessing basic clinical (the Richi articular index, total pain index, local articular index, pain index for knee and hand joints, circumference of knee and wrist joints) and ultrasound (the magnitude of exudate, an erosive process, osteophytes, articular fissure stenosis, the thickness of the synovial membrane and cartilage) indices. RESULTS: Use of OBT in patients with RA and OA just before articular radiation therapy brought about a more pronounced positive effect of complex therapy. In addition to significant positive changes in clinical parameters, there was a slow progression of a pathological process, as evidenced by ultrasound study. CONCLUSION: By reducing needs for drug therapy, for nonsteroidal antiflammatory drugs in particular, OBT produces a pharmacoeconomic effect. PMID- 12577852 TI - [Optimal drug therapy of glucose intolerance is a guarantee of successful prevention of type 2 diabetes mellitus]. PMID- 12577853 TI - [Diagnosis and treatment policy in chronic enteropathies]. PMID- 12577854 TI - [Nature of new markers of general pathology]. PMID- 12577856 TI - [Acute leukemia: which of two types of cell death (necrosis or apoptosis) may be responsible for patients' death during intensive chemotherapy?]. PMID- 12577855 TI - [Use of tanakan in disturbances of memory and attention in the elderly]. PMID- 12577858 TI - We've got to stop eating like this. PMID- 12577857 TI - Is fat the next tobacco? PMID- 12577859 TI - Uphill battle. PMID- 12577860 TI - The quest for the antifat pill. PMID- 12577861 TI - Sticker shock at the hospital. Freed from reins of managed care, hospitals accelerate price hikes for fourth consecutive year; blame placed on staffing, tech costs. AB - Free from the shackles of managed care, the nation's hospitals boosted prices for the fourth consecutive year, according to a new report released by the government last week. David Cyganowski, left, a managing director at Salomon Smith Barney, says if taxes on dividends are wiped out, hospitals could be confronted with higher long-term interest rates. PMID- 12577862 TI - N.J. docs set to launch walkout over escalating premium costs. Bush offers Rx for malpractice insurance crisis, including $250,000 limit on noneconomic damages. PMID- 12577863 TI - JCAHO, NCQA join forces. PMID- 12577864 TI - Fraud flurry. No connection seen in recent moves by government. PMID- 12577865 TI - Tort retort. Doc wins $30 million defamation suit against hospital. PMID- 12577866 TI - Health hazard. Medicaid cuts could endanger patients. PMID- 12577867 TI - Mergers minus the mania. Smaller deals rule as economic uncertainties contribute to a fourth annual decrease in hospital consolidation activity. PMID- 12577868 TI - FPM and the family physician's journey to medical excellence. PMID- 12577869 TI - Successful test-result tracking. PMID- 12577870 TI - Refills: clinical not clerical. PMID- 12577871 TI - CPT coding update for 2003. PMID- 12577872 TI - Medicare payment cuts yet to be corrected. PMID- 12577873 TI - Primary care patient safety research under way. PMID- 12577874 TI - Coding better for better reimbursement. PMID- 12577875 TI - Choosing between clinical practice & administration. PMID- 12577876 TI - Providing charity care: a primer on liability risk. PMID- 12577877 TI - Implementing change: from ideas to reality. PMID- 12577878 TI - Open access as an alternative to patient combat. PMID- 12577879 TI - 10 quick tips for a more user-friendly PDA. PMID- 12577880 TI - [Health care in today's Western Belarus in the inter-war period (1921-1939)]. AB - During the inter-war period, today's Western Belarus, which under the Treaty of Riga constituted part of the Second Polish Republic, stood out for its higher morbidity in various disease categories than other regions of the country. Heightened threats of epidemics prevailed in that area, and typhoid fever, trachoma, tuberculois and venereal disease were serious health problems. Specific features of the health care of today's Western Belarus could be observed that set it apart from both that of Eastern Belarus forming a part of the Soviet Union as well as from the central and western provinces of Poland. Compared with Eastern Belarus, there was a lack of health services, including anti-tuberculosis, anti venereal and paediatric dispensaries, or they were in the process of being created. But physicians' self-government bodies functioned, as did out-patient care with dispensary-like elements and health care provided by local health centres. There also existed the institution of full-time family doctors within the insured health-care system - something lacking in Eastern Belarus. As in other voivodships (provinces) of the Second Republic, there existed multi-sector health care: state, local, insurance-based and private. The Western Belarus had Poland's smallest number of physicians, new hospital beds and pharmacies. On the other hand, today's Western Belarus stands out by virtue of maintaining field medicine traditions. To this day, assistant medical officers provide health care to the rural population - an arrangement not typical of the central and western provinces fo pre-war Poland. PMID- 12577881 TI - [out of scope]. PMID- 12577882 TI - Doctors protest conscience discrimination: the Holy Father's address to the International Congress of Catholic Obstetricians and Gynaecologists, Rome, 18 June 2001. PMID- 12577883 TI - [out of scope]. PMID- 12577885 TI - [out of scope]. PMID- 12577884 TI - Protecting human life in a changing world: the responsibility of Catholic obstetricians. PMID- 12577886 TI - A question of conscience. PMID- 12577887 TI - The importance of the Catholic obstetrician-gynaecologist for the presence of the Church in the world of health and health care. PMID- 12577888 TI - Managing ethics under managed care. PMID- 12577890 TI - Neural fetal tissue transplants: old and new issues. AB - Neural fetal tissue transplantation offers promise as a treatment for devastating neurologic conditions such as Parkinson's disease. Two types of issues arise from this procedure: those associated with the use of fetuses, and those associated with the use of neural tissue. The former issues have been examined in many forums; the latter have not. This paper reviews issues and arguments raised by the use of fetal tissue in general, but focuses on the implications of the use of neural tissue for basic concepts of personhood and personal identity. PMID- 12577891 TI - Space life sciences: missions to Mars, radiation biology, and plants as a foundation for long-term life support systems in space. Refereed papers from the F0.1 and F1.3-F2.3 Symposia of COSPAR Scientific Commission F which were held during the Thirty-third COSPAR Scientific Assembly, Warsaw, Poland, July 2000. PMID- 12577892 TI - [Risk management in radiology: for growing safety culture and awareness for preventing malpractice]. PMID- 12577893 TI - Issues of exploration: human health and wellbeing during a mission to Mars. AB - Today, the tools are in our hands to enable us to travel away from our home planet and become citizens of the solar system. Even now, we are seriously beginning to develop the robust infrastructure that will make the 21st century the Century of Space Travel. But this bold step must be taken with due concern for the health, safety and wellbeing of future space explorers. Our long experience with space biomedical research convinces us that, if we are to deal effectively with the medical and biomedical issues of exploration, then dramatic and bold steps are also necessary in this field. We can no longer treat the human body as if it were composed of muscles, bones, heart and brain acting independently. Instead, we must lead the effort to develop a fully integrated view of the body, with all parts connected and fully interacting in a realistic way. This paper will present the status of current (2000) plans by the National Space Biomedical Research Institute to initiate research in this area of integrative physiology and medicine. Specifically, three example projects are discussed as potential stepping stones towards the ultimate goal of producing a digital human. These projects relate to developing a functional model of the human musculoskeletal system and the heart. PMID- 12577894 TI - The development of the physicians' end-of-life care attitude scale. AB - The Physicians' End-of-Life Care Attitude Scale (PEAS) was developed as an outcome measure for palliative care education. PEAS assesses the willingness of medical trainees to care for dying patients. Sixty-four Likert-type questions were created on the basis of discussions with focus groups of medical trainees, then administered to sixty-two medical students and residents. Total PEAS scores as well as personal preparation and professional role subscales (where higher scores indicated greater concern) possessed excellent internal consistency and reliability. In addition, there were substantial correlations between PEAS scores and the CA-Dying scale, a measurement of laypersons' fears about interacting with dying persons. Thus, PEAS adequately assesses the unique communication concerns of physicians in training regarding working with dying persons and their families. Correlations between PEAS scores and age were negative, while those who had experienced the death of a loved one had higher PEAS scores than those who did not. This suggests that for some persons, life experiences may lessen difficulties in dealing with dying persons, while for others, personal losses may exacerbate such concerns. The utility of PEAS in evaluating the efficacy of palliative care education as well as its potential to measure medical trainee's willingness to care for the terminally ill is discussed. PMID- 12577895 TI - [Herbert's blue eyes]. PMID- 12577896 TI - An alternative approach to solar system exploration providing safety of human mission to Mars. AB - For systematic human Mars exploration, meeting crew safety requirements, it seems perspective to assemble into a spacecraft: an electrical rocket, a well-shielded long-term life support system, and a manipulator-robots operating in combined "presence effect" and "master-slave" mode. The electrical spacecraft would carry humans to the orbit of Mars, providing short distance (and low signal time delay) between operator and robot-manipulators, which are landed on the surface of the planet. Long-term hybrid biological and physical/chemical LSS could provide environment supporting human health and well being. Robot-manipulators operating in "presence effect" and "master-slave" mode exclude necessity of human landing on Martian surface decreasing the level of risk for crew. Since crewmen would not have direct contact with the Martian environment then the problem of mutual biological protection is essentially reduced. Lightweight robot-manipulators, without heavy life support systems and without the necessity of returning to the mother vessel, could be sent as scouts to different places on the planet surface, scanning the most interesting for exobiological research site. Some approximate estimations of electric spacecraft, long-term hybrid LSS, radiation protection and mission parameters are conducted and discussed. PMID- 12577897 TI - Atharvaveda and its materia medica. AB - Atharvaveda is the fourth and last Veda of Hindu literature. Its oldest name was 'ATHARVANGIRASAH', because it was contributed by two sages, ATHARVAN and ANGIRA. It is also known as 'Bhaishajjvaveda'. Atharvaveda gives information regarding plants, minerals and animal products with their usage for medical purposes. For example, 'Apamarga', a plant is useful for cough, piles, itching and abdominal pain, wherea 'Lavana' is useful for pimples; 'Shankha' useful to protect from diseases and 'Mriga Shringa' is useful for pulmonary consumption and other chronic diseases etc. PMID- 12577898 TI - Introduction. PMID- 12577899 TI - The menace of free radicals its concept and management in Ayurveda. AB - This is a comprehensive review of menace of free radicals and its concept and management in Ayurveda. This article highlights the various exogenous and endogenous factors responsible for the production of free radicals with special reference to the formation of unriped and purified metabolites (ama) during the metabolic activities at different levels of digestion. The purpose of this paper is to review the management of free radicals which can be designed with reference to diet and digestion biorhythms, behaviour, emotions and sense. Ayurvedic concept of free radicals scavenger, revitalisation, role of clarified butter and pollution control thereby maintaining the optimal health status by involving minimum of sophisticated technology thus guiding the individual to lead a harmonious life in unison with nature. PMID- 12577900 TI - Benefits from human presence to the exploration of Mars. PMID- 12577901 TI - Who decides? The connecting thread of euthanasia, eugenics, and doctor-assisted suicide. AB - Throughout recorded history, a series of seemingly unrelated ideas have been consistently intertwined: suicide, euthanasia, infanticide, eugenics, genocide and, most recently, the practice termed physician-assisted suicide. From Plato and Hippocrates to a pair of twentieth-century American physicians named Haiselden and Kevorkian, an examination of history shows these disparate notions always involve two troublesome questions: Which lives are not worth living? And who will decide? The same examination of history teaches that separating the worthy from the not worthy is a very dangerous proposition, especially for those whose lives are deemed marginal. PMID- 12577902 TI - [How your colleagues successfully recruit patients for their practice]. PMID- 12577903 TI - Radiation risk and human space exploration. AB - Radiation protection is essential to enable humans to live and work safely in space. Predictions about the nature and magnitude of the risks posed by space radiation are subject to very large uncertainties. Prudent use of worst-case scenarios may impose unacceptable constraints on shielding mass for spacecraft or habitats, tours of duty of crews on Space Station, and on the radius and duration of sorties on planetary surfaces. The NASA Space Radiation Health Program has been devised to develop the knowledge required to accurately predict and to efficiently manage radiation risk. The knowledge will be acquired by means of a peer-reviewed, largely ground-based and investigator-initiated, basic science research program. The NASA Strategic Plan to accomplish these objectives in a manner consistent with the high priority assigned to the protection and health maintenance of crews will be presented. PMID- 12577904 TI - Hippocrates, Maimonides and the doctor's responsibility. AB - Jack Kevorkian criticizes the Hippocratic tradition in Greek medicine, which bans the physician from giving his patient a lethal medication. He sees this prohibition as potentially bringing harm to a suffering patient and not reflective of the larger Greek society which was tolerant and even approving of suicide. However, Kevorkian's advocacy of doctor-assisted suicide can be seen as the polarity of doctor abandonment of the suffering patient rather than as an antidote to it. Both positions involve an outcome of physician removal from the suffering patient, which can be contrasted with Maimonides' command to the physician to watch over the life and death of his patients. PMID- 12577905 TI - [Assessing plausibility of accusations of double treatment in group practice]. PMID- 12577906 TI - Diagnostic endomyocardial biopsy interpretation. PMID- 12577907 TI - [Changes in the concept of chronic pancreatitis and its future: commemorating the 100th year since the establishment of the Japanese Society of Pancreatology]. PMID- 12577908 TI - Gender, pain, and doctor involvement: high school student attitudes toward doctor assisted suicide. AB - The present study concentrates on the attitudes of high school students toward active doctor-assisted suicide as described in hypothetical doctor-patient scenarios, orthogonally manipulating doctor's reaction to patient's wishes to end his/her life (whether discussed, accepted or encouraged), presence of patient's physical pain, presence of patient's emotional pain, and the gender of the hypothetical patient. Doctor-assisted suicides thoroughly discussed with the patient are judged to be more moral, acceptable, and "legal" than assisted suicides that are simply accepted by the doctor or actively encouraged by him. Significantly, this is not a distinction that is relevant in the eyes of the law. Further, the presence of both physical and emotional pain on the part of the patient make the patient death more acceptable in the eyes of high school students. This latter effect is striking, given the result of the Wooddell and Kaplan (1999-2000) study showing that patient depression tends to weaken acceptability of death. Finally, respondents, both male and female, tend to view deaths of patients of the opposite gender as more acceptable than patients of the same gender. PMID- 12577910 TI - [History of internal medicine in the last 100 years]. PMID- 12577909 TI - Kaya Kalpa methods adopted by Siddars: a scientific approach. AB - Thirumular says in his "THIRUMANTHIRAM" the body is a sacred instrument to contain the soul and therefore must be nurtured well to safeguard the life. Thirumular is reported to have lived a long span of three thousand years according to tradition. The art of longevity and the attempts of immortalising the corporeal human body were the ultimate aims of Siddhars as understood from their numerous treatises on the art of rejuvenation - Thirumular Karpam - 300, Bohar Karpam - 300, Thiruvalluvur Karpam - 300 Yugimuni Karpam - 300 etc. which form some of the excellence works in this regard. One of the means to rejuvenate the body was achieved through the art of breathing (Pranayama) which has a direct bearing on the basal metabolic rate and the span of longevity. In this paper an attempt has been made to throw some light on the scientific basis of the various Kaya Kalpa methods adopted by Siddhars. PMID- 12577911 TI - Effect of the doctor on college students' attitudes toward physician-assisted suicide. AB - Ninety-six students were presented with eighteen different vignettes describing different types of active and passive observed suicide, assisted suicide, and euthanasia. Attitudes regarding the morality and desired legality of each situation were measured. Results indicated that the interaction between the doctor and the patient, and, to a lesser extent, the active or passive nature of the agent of death, were more important than the actual actions of the doctor in allowing or causing death to occur. PMID- 12577912 TI - New guidelines, research add to mammography controversy. PMID- 12577913 TI - Drug mistakes are very common, study says. PMID- 12577914 TI - [Discovery of a new physiopathological aspect of arteriosclerosis and elucidation of its molecular mechanism]. PMID- 12577915 TI - Doctor assisted suicide: an analysis of public opinion of Michigan adults. AB - Michigan public opinion on Doctor Assisted Suicide (DAS) was assessed in January 1997 (N=603). Asked if they would consider DAS for themselves, two-thirds would if being kept alive by machine or were experiencing chronic pain; one-half would if they experienced a loss in mobility or independence, became a burden to others, or were diagnosed with a terminal disease; and one-third would if they were incontinent or going to a nursing home. A series of demographic and attitudinal comparisons were made for support for the concept of DAS and as a hypothetical consideration for oneself. The highest support for the concept of DAS was found among the following: men eighteen to twenty-four years old, some college education, $35-60,000/year income, Caucasian, Democrat, liberal, Protestant, and frequent church attendee. The highest self-consideration of DAS was found among the following: men, fifty to fifty-five years old, post-graduate education, $35-60,000/year income, Caucasian, Democrat, liberal, Protestant, and infrequent church attendee. PMID- 12577916 TI - Radiation environment monitoring for manned missions to Mars. AB - In this paper a radiation monitoring system for manned Mars missions is described, based on the most recent requirements on crew radiation safety. A comparison is shown between the radiation monitoring systems for Earth-orbiting and interplanetary spacecraft, with similarities and differences pointed out and discussed. An operational and technological sketch of the chosen problem solving approach is also given. PMID- 12577917 TI - [From anti-arrhythmia drugs to therapeutic agents for heart failure]. PMID- 12577918 TI - Diabetics who see specialists have lower rates of long-term complications. PMID- 12577919 TI - Occupational therapy is cost-effective and improves quality of life for seniors. PMID- 12577920 TI - Personal dosimetry for human missions to Mars based on TLD and LET-spectrometry technique. AB - Exposure of crew to the space radiation environment poses one of the most significant problems in long term missions in low earth orbits and in interplanetary missions. Accurate personal dose measurement will become increasingly important especially during manned missions to Mars. A series of instruments suitable for on-board dose, flux and LET measurements has been developed by the authors'. Based on the experience gained so far from their utilization, an instrument consisting of a thermoluminescent device and a dosimetry telescope is proposed for the use on the International Space Station and for human Mars missions. The short technical description of this instrument is given in this paper. PMID- 12577921 TI - [Control of respiratory tract diseases of the aged]. PMID- 12577922 TI - Abstracts of the annual meeting of the Australasian Society for Infectious Diseases. Barossa Valley, South Australia, Australia. 13-17 April 2002. PMID- 12577923 TI - [Urine concentrating mechanism of the kidney and membrane transport anomalies]. PMID- 12577924 TI - Comment on estimating the solar proton environment that may affect Mars missions. AB - Estimates of the energetic proton environment for a Mars mission are generally extrapolated from the solar proton observations at 1 AU. We find that solar particle events may be divided into two general classes. Events dominated by a near-sun injection of particles onto interplanetary magnetic field lines leading to the spacecraft position represent the "classical" solar particle event associated with solar activity. This class of event will scale in radial distance by the classical power law extrapolation. The extended-interplanetary-shock source generates a maximum flux as the shock passes the detection location. This class of event typically generates maximum fluence, but in this case, the flux and fluence will not scale in the classical manner with radial distance. PMID- 12577925 TI - [Mechanism responsible for the development of acute liver failure and its management]. PMID- 12577926 TI - Life support approaches for Mars missions. AB - Life support approaches for Mars missions are evaluated using an equivalent system mass (ESM) approach, in which all significant costs are converted into mass units. The best approach, as defined by the lowest mission ESM, depends on several mission parameters, notably duration, environment and consequent infrastructure costs, and crew size, as well as the characteristics of the technologies which are available. Generally, for the missions under consideration, physicochemical regeneration is most cost effective. However, bioregeneration is likely to be of use for producing salad crops for any mission, for producing staple crops for medium duration missions, and for most food, air and water regeneration for long missions (durations of a decade). Potential applications of in situ resource utilization need to be considered further. PMID- 12577927 TI - [Molecular physiopathology of gastric precancerous conditions]. PMID- 12577928 TI - New problems to be solved for establishing closed life support system. AB - New test bed facilities such as Bioplex and CEEF have been constructed to test the new advanced technologies for solving the various problems as follows, (1) how to develop air content stabilization technologies with gas balance between the generation and the absorption by living organisms, (2) how to solve the mismatching between the assimilation rate of autotrophic organisms and the respiration rate of heterotrophic organisms, (3) how to balance the speed of the waste decomposition with the absorption speed of nutrient components in the sequential plant cultivation, (4) how to develop new nutrient adjusting subsystems for each plant species, (5) how to compensate the denitrification during the waste decomposition and anaerobic microbes in the nutrient solution. PMID- 12577929 TI - [Molecular biological study of physiopathology of inflammatory bowel diseases (Crohn disease and ulcerative colitis)]. PMID- 12577930 TI - [Molecular physiopathology and clinical aspect of pancreatitis]. PMID- 12577931 TI - Physician-assisted suicide and euthanasia: the pharmacist's perspective. AB - Pharmacists are in a critical position when pharmaceutical agents are prescribed for the purpose of physician-assisted suicide and/or euthanasia and they may need to decide whether dispensing a lethal dose of a medication is ethically and morally acceptable for a patient. In many cases, pharmacists may not even be aware that prescriptions are intended for physician-assisted suicide and/or euthanasia. Pharmacists have a special responsibility to protect patients who are contemplating end-of-life decisions such as physician-assisted suicide and euthanasia. Pharmaceutical care ("Responsible provision of drug therapy for the purpose of achieving definite outcomes that improve a patient's quality of life ...") requires that the pharmacist not only understands the medications but also the individual patient and the complexities of their lives and suffering. Only in this way can pharmacists provide safe and effective use of medications for the patients they serve. PMID- 12577932 TI - Access to specialists: perspectives of patients and primary care physicians. PMID- 12577933 TI - [Molecular physiopathology of viral hepatitis]. PMID- 12577934 TI - Development and research program for a soil-based bioregenerative agriculture system to feed a four person crew at a Mars base. AB - For humans to survive during long-term missions on the Martian surface, bioregenerative life support systems including food production will decrease requirements for launch of Earth supplies, and increase mission safety. It is proposed that the development of "modular biospheres"--closed system units that can be air-locked together and which contain soil-based bioregenerative agriculture, horticulture, with a wetland wastewater treatment system is an approach for Mars habitation scenarios. Based on previous work done in long-term life support at Biosphere 2 and other closed ecological systems, this consortium proposes a research and development program called Mars On Earth(TM) which will simulate a life support system designed for a four person crew. The structure will consist of 6 x 110 square meter modular agricultural units designed to produce a nutritionally adequate diet for 4 people, recycling all air, water and waste, while utilizing a soil created by the organic enrichment and modification of Mars simulant soils. Further research needs are discussed, such as determining optimal light levels for growth of the necessary range of crops, energy trade offs for agriculture (e.g. light intensity vs. required area), capabilities of Martian soils and their need for enrichment and elimination of oxides, strategies for use of human waste products, and maintaining atmospheric balance between people, plants and soils. PMID- 12577935 TI - Potential psychodynamic factors in physician-assisted suicide. AB - A number of assumptions underlying the debate over physician-assisted suicide (PAS) deserve closer scrutiny. It is often implicitly assumed that decisions as to the competency of the patient to request PAS can be accurately made, and that the treating physician's values and intrapsychic conflicts can be successfully separated from the decision to accede to or reject the patient's request. This article argues that in such an emotionally-laden decision, such factors may play a significant role, and that even were PAS to gain widespread acceptance, ignoring them may lead to errors in classifying patients either as appropriate or inappropriate for PAS. PMID- 12577936 TI - Psychosocial versus biomedical risk factors in Kevorkian's first forty-seven physician-assisted deaths. AB - This article examines biomedical and psychosocial data on the first forty-seven cases of physician-assisted suicide (PAS) of Kevorkian as collected by means of both a physical autopsy and a preliminary psychological autopsy. The following patterns emerge: 1) The physical condition of these PAS patients was not typical of the conditions that lead to death in the United States. 2) Consistent with the above findings, our pilot data indicate that only 31.1 percent of these patients were terminal. While 73.9 percent were described as reporting pain, only 42.6 percent were revealed at autopsy to have a specific anatomical basis for their pain. However 36 percent were described as depressed, 66 percent as having some disability, and perhaps of key importance, 90 percent expressed a fear of dependency. Most important, our pilot data suggest the possibility of large gender differences, since 3) 68.1 percent of these forty-seven PAS's are women and only 31.9 percent are men. This represents the reverse of the gender pattern for completed suicides in the United States in 1995, resembling instead the approximate pattern for unsuccessful suicide attempts. 4) Approximately 75 percent of both men and women in the above sample were described as reporting pain. Men were almost twice as likely to have had an anatomical basis for the pain and three times as likely to be terminal. Our pilot data indicate PAS women are more likely to be described as depressed and twice as likely to have had a history of previous unsuccessful suicide attempts. 5) Kevorkian's patients were older than the typical unaided suicides in America. Reported pain decreases with age as does depression; however anatomical basis for pain increases slightly with age, and no age effect emerges for terminality. 6) Approximately two-thirds of those physician-assisted suicides were at middle SES levels. History of disability was the biggest risk factor for the low SES patients and fear of dependency for the high SES patients. PMID- 12577937 TI - Satisfaction and quality: patient and physician perspectives. PMID- 12577938 TI - [Molecular cytogenetic analysis of cancer of the digestive system and its clinical application]. PMID- 12577939 TI - Possible applications of aquatic bioregenerative life support modules for food production in a Martian base. AB - Water is the essential precondition of life in general and also for the establishment of a Martian base suitable for long duration stays of humans. It is not yet proven if there is indeed a "frozen ocean" under the surface of Mars but if this could be verified it would open innovative aspects for the construction of bioregenerative life support systems (BLSS). In a general concept higher plants will play the predominant role in a Martian BLSS. It is not clear, however, how these will grow and bring seed in reduced gravity and there may be differences in the productivity in comparison to Earth conditions. Therefore, organisms which are already adapted to low gravity conditions, namely non gravitropic aquatic plants and also aquatic animals may be used to enhance the functionality of the Martian BLSS as a whole. It has been shown already with the so-called C.E.B.A.S. MINIMODULE in the STS-89 and STS-90 spaceshuttle missions that the water plant Ceratophyllum demersum has an undisturbed and high biomass production under space conditions. Moreover, the teleost fish species Xiphophorus helleri adapted easily to the micro-g environment and maintained its normal reproductive functions. Based on this findings a possible scenario is presented in which aquatic plant production modules and combined animal-plant production systems may be used for human food production and water and air regeneration in a Martian base. PMID- 12577940 TI - Dissatisfaction with consent for diagnostic gastrointestinal endoscopy. AB - INTRODUCTION: Little is known about patients' perspectives on the amount of information they receive prior to diagnostic gastrointestinal endoscopies. Our unit's policy for obtaining consent consists of initially posting an information leaflet to the patient followed by subsequent explanation of the procedure on arrival for the test. The consent form is signed by the patient immediately prior to the test. METHODS: A questionnaire survey was conducted to assess patient perception and satisfaction with the amount of information received before diagnostic endoscopy. RESULTS: The information was obtained from 127 of the 175 questionnaires that were distributed. Whereas 97% had read the information leaflet, only 52% had read the consent form before signing it. 64/127(51%) felt dissatisfied because they would have wanted more information while 3% were dissatisfied because they would have liked less information relating to one or more aspects of the test. Dissatisfaction was higher in patients who had not read the consent form (p < 0.001) and those with some formal education (p = 0.01). CONCLUSIONS: Patients who did not read the consent form were more dissatisfied. Strategies to improve the rate of reading this document may increase patient satisfaction. PMID- 12577941 TI - Access to care: is it improving or declining? PMID- 12577942 TI - Informed consent for gastrointestinal endoscopy: a 2002 ESGE survey. AB - BACKGROUND/AIM: Gastroenterologists are required to obtain informed consent before undertaking any endoscopic examination. Published data indicate that in practice there are many deficiencies in this process. The aim of this survey was to determine the quality of information given to patients before the endoscopic procedures in countries that are members of the European Society of Gastrointestinal Endoscopy (ESGE). METHODS: A structured questionnaire was sent to representatives of endoscopic societies that are members of ESGE regarding the quality of informed consent. RESULTS: The response rate was 59% (26/44). The endoscopist is responsible for giving the required information prior to written consent in only 23.1% (6/26) of the countries. Although information about the procedure is given to the patients in 96% of the responding countries, in only 77% is there sufficient time for the patients to ask questions about the nature of the procedure. In 15% (4/26) of the countries neither the diagnostic or therapeutic alternatives to endoscopy are discussed nor the potential complication rates. Finally, the procedure-related mortality is provided in only 23% (6/26) of countries. CONCLUSION: The procedure of obtaining informed consent for endoscopy varies considerably and may need reevaluation. PMID- 12577943 TI - Assessment of depression in patients with chronic hepatitis: effect of interferon treatment. AB - INTRODUCTION/AIM: Chronic viral hepatitis and its treatment with interferon (IFN) have an impact on patients' quality of life. The purpose of this study was to evaluate the degree of depression in patients with chronic viral hepatitis before and during IFN therapy. PATIENTS/METHODS: Our study group comprised 132 patients, 38 with chronic hepatitis C (26 males, 12 females), 36 with chronic hepatitis B (29 males, 7 females), and 58 individuals with no chronic disease (24 males, 34 females). Depression was assessed using the Zung Self-Rating Depression Scale and the corresponding SDS index. RESULTS: At baseline, patients with viral hepatitis and healthy controls had similar SDS indexes. During treatment, a significant increase in the SDS index was observed in both HBV and HCV patients (p < 0.001), which returned to the baseline levels after treatment. During IFN therapy, the SDS indexes were higher in HCV compared to HBV patients and women compared to men (p = 0.001 and p = 0.008, respectively), and there was a >5- fold increased frequency to develop severe depression. CONCLUSIONS: IFN-induced depression occurs more frequently in HCV than HBV patients and in women than men. Monitoring during treatment is mandatory in order to maximize treatment adherence. PMID- 12577944 TI - Gender and physician-assisted suicide: an analysis of the Kevorkian cases, 1990 1997. AB - This study examines the seventy-five suicide cases Dr. Jack Kevorkian acknowledged assisting during the period between 1990-1997. Although these cases represent a range of regional and occupational backgrounds, a significant majority are women. Most of these individuals had a disabling, chronic, nonterminal-stage illness. In five female cases, the medical examiner found no evidence of disease whatsoever. About half of the women were between the ages of forty-one and sixty, and another third were older adults. Men's conditions were somewhat less likely than women's to be chronic and nonterminal-stage. The main reasons for the hastened death mentioned by both the person and their significant others were having disabilities, being in pain, and fear of being a burden. The predominance of women among Kevorkian's assisted suicides contrasts with national trends in suicide mortality, where men are a clear majority. It is possible that individuals whose death was hastened by Kevorkian are not representative of physician-assisted suicide cases around the country, because of Kevorkian's unique approach. Alternatively, the preponderance of women among Kevorkian's assisted suicides may represent a real phenomenon. One possibility is that, in the United States, assisted suicide is particularly acceptable for women. Individual, interpersonal, social, economic, and cultural factors encouraging assisted suicide in women are examined. PMID- 12577945 TI - [Molecular mechanism and biology of neovascularization]. PMID- 12577946 TI - Percutaneous endoscopic gastrostomy: adequacy and quality of information given to decision makers. AB - BACKGROUND: Nowadays percutaneous endoscopic gastrostomy (PEG) is widely available, but patient-selection criteria and quality of informed consent are debated. The aims of this retrospective study were to evaluate the quality of information given to the decision-makers (relatives) and determine the overall acceptance of the procedure by the patients' family. METHODS: The relatives of patients with PEG were interviewed by telephone, using a structured questionnaire. They (n = 55; 36% spouses, 34% children, 30% other) gave information about themselves and the patient (34 males, 21 females, median age 69, range 16-92 years) who underwent PEG tube placement for eating disorders or dysphagia. RESULTS: At the time of evaluation 30/55 (54.6%) patients had died. The cumulative median survival was significantly longer in patients younger than 75 years by 58 days (p = 0.009). Relatives believed that PEG could improve the patients' quality of life (56%) or/and the underlying disease. Although 93% of the decision-makers considered that their opinion had been taken into account when the procedure was done, 25% said that they had not adequately been informed about alternative methods and the complications of the procedure (38%). 54% said that the procedure had improved the quality of life of the family. Most of the decision-makers believed that their decision was correct (87%) and they would recommend PEG (84%) to other patients suffering from dysphagia. CONCLUSION: Though several decision-makers were not satisfied with the quality of information given before informed consent, the overall acceptance of the PEG placement for nutritional support is high. PMID- 12577947 TI - Strong opinions held about the tradeoff between choice of providers and cost of care. PMID- 12577948 TI - Critical issues in connection with human missions to Mars: protection of and from the Martian environment. AB - Human missions to Mars are planned to happen within this century. Activities associated therewith will interact with the environment of Mars in two reciprocal ways: (i) the mission needs to be protected from the natural environmental elements that can be harmful to human health, the equipment or to their operations; (ii) the specific natural environment of Mars should be protected so that it retains its value for scientific and other purposes. The following environmental elements need to be considered in order to protect humans and the equipment on the planetary surface: (i) cosmic ionizing radiation, (ii) solar particle events; (iii) solar ultraviolet radiation; (iv) reduced gravity; (v) thin atmosphere; (vi) extremes in temperatures and their fluctuations; and (vii) surface dust. In order to protect the planetary environment, the requirements for planetary protection as adopted by COSPAR for lander missions need to be revised in view of human presence on the planet. Landers carrying equipment for exobiological investigations require special consideration to reduce contamination by terrestrial microorganisms and organic matter to the greatest feasible extent. Records of human activities on the planet's surface should be maintained in sufficient detail that future scientific experimenters can determine whether environmental modifications have resulted from explorations. PMID- 12577949 TI - Primary care physicians concerned about patients' access to mental health services. PMID- 12577950 TI - Gatekeeping arrangements are in widespread use. PMID- 12577951 TI - Payment arrangements and financial incentives for physicians. PMID- 12577952 TI - [Rheumatoid arthritis and neovascularization]. PMID- 12577953 TI - Nonsteroidal anti-inflammatory drugs and the risk of intracerebral hemorrhage. PMID- 12577954 TI - [Etiology of diabetic retinopathy]. PMID- 12577955 TI - Trends in managed care coverage in small firms. PMID- 12577956 TI - [Cardiovascular diseases and neovascularization]. PMID- 12577957 TI - Growth of Medicaid-dominated HMOs. PMID- 12577958 TI - [Multiple myeloma and neovascularization]. PMID- 12577959 TI - How widespread is managed competition? PMID- 12577960 TI - Sterilisation properties of the Mars surface and atmospheric environment. AB - The radiative and chemical conditions at the surface and in the lower Martian atmosphere are computed at various latitudes and seasons combining a 2D photochemical model and radiation simulations. In most situations, the solar UV B and C radiations reach the surface however, suspended dust and, in polar cases, ozone can constitute an effective UV shield. The daytime and night time concentrations of the sterilizing oxidants: OH, H2O2 and O3 are determined, as well as the concentration of the substances which could influence the metabolism of microorganisms. The possible habitats of a remaining Mar's life as well as the possibilities of contamination by resistant earth life forms will be described. PMID- 12577961 TI - [Development of lung cancer and neovascularization]. PMID- 12577962 TI - Despite fears, costs rise only modestly in 1998. PMID- 12577963 TI - [Epidemiology of dementia--from the Hisayama study]. PMID- 12577964 TI - Trends in the cost of employer-sponsored coverage. PMID- 12577965 TI - Planetary protection and the search for life beneath the surface of Mars. AB - The search for traces of extinct and extant life on Mars will be extended to beneath the surface of the planet. Current data from Mars missions suggesting the presence of liquid water early in Mars' history and mathematical modeling of the fate of water on Mars imply that liquid water may exist deep beneath the surface of Mars. This leads to the hypothesis that life may exist deep beneath the Martian surface. One possible scenario to look for life on Mars involves a series of unmanned missions culminating with a manned mission drilling deep into the Martian subsurface (approximately 3Km), collecting samples, and conducting preliminary analyses to select samples for return to earth. This mission must address both forward and back contamination issues, and falls under planetary protection category V. Planetary protection issues to be addressed include provisions stating that the inevitable deposition of earth microbes by humans should be minimized and localized, and that earth microbes and organic material must not contaminate the Martian subsurface. This requires that the drilling equipment be sterilized prior to use. Further, the collection, containment and retrieval of the sample must be conducted such that the crew is protected and that any materials returning to earth are contained (i.e., physically and biologically isolated) and the chain of connection with Mars is broken. PMID- 12577966 TI - Labeling requirements for systemic antibacterial drug products intended for human use. Final rule. AB - The Food and Drug Administration (FDA) is amending its regulations to require that the labeling for all systemic antibacterial drug products (i.e., antibiotics and their synthetic counterparts) intended for human use include certain statements about using antibiotics in a way that will reduce the development of drug-resistant bacterial strains. The final rule reflects a growing concern in FDA and the medical community that unnecessary use of systemic antibacterials has contributed to a dramatic increase in recent years in the prevalence of drug resistant bacterial infections. The final rule is intended to encourage physicians to prescribe systemic antibacterial drugs only when clinically necessary. The final rule is also intended to encourage physicians to counsel their patients about the proper use of such drugs and the importance of taking them exactly as directed. PMID- 12577967 TI - Exercise testing in African American youth. PMID- 12577968 TI - [Clinical description of vascular dementia]. PMID- 12577969 TI - The big picture. PMID- 12577970 TI - Trends in offering employer-sponsored coverage. PMID- 12577971 TI - Physicians and care management: more acceptance than you think. AB - Physicians' acceptance of care management tools--practice guidelines, patient satisfaction surveys and practice profiling--is an important indication of whether these efforts to improve quality and control health costs will succeed. According to a new study by the Center for Studying Health System Change (HSC), a majority of physicians affected by these tools believe the overall effect of each on the quality and efficiency of care they provide has been positive. Physicians affected by related financial incentives are more likely to view care management techniques positively if they are risk adjusted to reflect the greater need for services by people with health problems. PMID- 12577972 TI - Effect of heavy ions on neuro-endocrine regulations. AB - During American and Russian short and long-term space flights neuroimmune dysregulations have been observed in man and rats for up to three months after the return. During Extra-Vehicular Activity, radiation exposure risk is greater to elicit short and/or long-term deleterious effects on the functional capacity of the neuroimmune system. In order to assess the effects of high LET events on neuroimmune networks, our preliminary ground-based study was to investigate brain inflammatory responses in mouse after low dose radiation exposure with high LET particles (12C, 95MeV/u, 42 mGy). Plasma corticosterone levels were rapidly (6 hours) increased by two-fold, then decreased 24 hours post-irradiation. At 3 days plasma corticosterone and ACTH concentrations were also two- to three-fold increased. Plasma ACTH levels were still elevated up to seven days to two months. Furthermore immune functions are under current assessment. The results of this study should allow a greater understanding of the effects of high LET particles on neuroimmune system. PMID- 12577973 TI - Why people change their health care providers. AB - This Data Bulletin presents findings from the Household Survey, a nationally representative telephone survey of the civilian, non-institutionalized population conducted as part of the Community Tracking Study. The 1996-1997 survey includes nearly 33,000 families and 60,000 individuals, while the 1998-1999 survey includes approximately 32,000 families and 59,000 individuals. PMID- 12577974 TI - Paying for family leave. AB - California was the first to pass a paid family leave law. Now other states are considering following in the Golden State's footsteps. PMID- 12577975 TI - [Clinical course of Alzheimer disease]. PMID- 12577976 TI - The genetic key to public health. AB - Strides in genetics research are making a difference in public health. PMID- 12577977 TI - Pharmacy Plus: the perfect cure? AB - States are experimenting with using Medicaid to cover drug costs for the "not quite-so-poor." PMID- 12577978 TI - Patients concerned about insurer influences. AB - This Data Bulletin presents findings from two rounds of the Household Survey, a nationally representative telephone survey of the civilian, noninstitutionalized population conducted as part of the Community Tracking Study. The 1996-1997 survey includes nearly 33,0000 families and 60,000 individuals, while the 1998 1999 survey includes about 32,000 families and 59,000 individuals. This analysis does not include people who lack health insurance. All comparisons and differences described are statistically significant as the p<.05 level. PMID- 12577979 TI - [Molecular mechanism of Alzheimer disease]. PMID- 12577980 TI - Who is likely to switch health plans? AB - This Data Bulletin presents findings from the 1996-1997 Household Survey, a nationally representative telephone survey of the civilian, noninstitutionalized population conducted as part of the Community Tracking Study. The 1996-1997 survey includes nearly 33,000 families and 60,000 individuals. PMID- 12577981 TI - Cognitive deficits induced by 56Fe radiation exposure. AB - Exposing rats to particles of high energy and charge (e.g., 56Fe) disrupts neuronal systems and the behaviors mediated by them; these adverse behavioral and neuronal effects are similar to those seen in aged animals. Because cognition declines with age, and our previous study showed that radiation disrupted Morris water maze spatial learning and memory performance, the present study used an 8 arm radial maze (RAM) to further test the cognitive behavioral consequences of radiation exposure. Control rats or rats exposed to whole-body irradiation with 1.0 Gy of 1 GeV/n high-energy 56Fe particles (delivered at the alternating gradient synchrotron at Brookhaven National Laboratory) were tested nine months following exposure. Radiation adversely affected RAM performance, and the changes seen parallel those of aging. Irradiated animals entered baited arms during the first 4 choices significantly less than did controls, produced their first error sooner, and also tended to make more errors as measured by re-entries into non baited arms. These results show that irradiation with high-energy particles produces age-like decrements in cognitive behavior that may impair the ability of astronauts to perform critical tasks during long-term space travel beyond the magnetosphere. PMID- 12577982 TI - SAH-induced cerebral vasospasm: unraveling molecular mechanisms of a complex disease. PMID- 12577983 TI - Citation for the Down Surgical Prize 2002, Michael Corrigan. PMID- 12577984 TI - Long-term changes in amphetamine-induced reinforcement and aversion in rats following exposure to 56Fe particle. AB - Exposing rats to heavy particles produces alterations in the functioning of dopaminergic neurons and in the behaviors that depend upon the integrity of the dopaminergic system. Two of these dopamine-dependent behaviors include amphetamine-induced reinforcement, measure using the conditioned place preference procedure, and amphetamine-induced reinforcement, measured using the conditioned place preference procedure, and amphetamine-induced aversion, measured using the conditioned taste aversion. Previous research has shown that exposing rats to 1.0 Gy of 1GeV/n 56Fe particles produced a disruption of an amphetamine-induced taste aversion 3 days following exposure, but produced an apparent enhancement of the aversion 112 days following exposure. The present experiments were designed to provide a further evaluation of these results by examining taste aversion learning 154 days following exposure to 1.0 Gy 56Fe particles and to establish the convergent validity of the taste aversion results by looking at the effects of exposure on the establishment of an amphetamine-induced conditioned place preference 3, 7, and 16 weeks following irradiation. The taste aversion results failed to confirm the apparent enhancement of the amphetamine-induced CTA observed in the prior experiment. However, exposure to 56Fe particles prevented the acquisition of amphetamine-induced place preference at all three-time intervals. The results are interpreted as indicating that exposure to heavy particles can produce long-term changes in behavioral functioning. PMID- 12577985 TI - Citation on behalf of Ian Martin for BAOMS Surgery Prize 2002. PMID- 12577986 TI - Study of the radiation environment on MIR space station with SILEYE-2 experiment. AB - In this work we present preliminary results of nuclear composition measurements on board space station MIR obtained with SILEYE-2 particle telescope. SILEYE-2 was placed on MIR in 1997 and has been working since then. It consists of an array of 6 active silicon strip detectors which allow nuclear and energetic identification of cosmic rays in the energy range between approximately 30 and 200 MeV/n. The device is attached to an helmet and connected to an eye mask which shields the cosmonaut eyes from light and allow studies of the Light Flashes (LF) phenomenon. In addition to the study of the causes of LF, the device is used to perform real time long term radiation environment monitoring inside the MIR, performing measurements in solar quiet and active days. PMID- 12577987 TI - Falls and injuries after stroke: time for action! PMID- 12577988 TI - Alauddin Qarshi: a commentator of many classical treatises. AB - This article deals with the life and work of a commentator of six authentic works. He was a discoverer of pulmonary circulation of blood. He described it more or less accurately, almost three centuries before Harvey (1578-1657). The views of medical historians about this scholar have been given in this article. PMID- 12577989 TI - ID NMR Methods in ligand-receptor interactions. AB - The drug discovery process often involves the screening of compound libraries to identify drug candidates capable of binding to target macromolecules. New approaches in biological and chemical research are driving a change in the pharmaceutical industry. Recent advances in NMR spectroscopy such as affinity NMR techniques, which detect binding of a small molecule with a "receptor", have been shown to be valuable tools to perform rapid screening of compounds for biological activity. These NMR observable events include using relaxation, chemical shift perturbations, translational diffusion, and magnetization transfer. These one dimensional NMR methods increase both the throughput of screening and yield crucial data on the mode of binding. The practical utility of these techniques will be described. PMID- 12577990 TI - NMR methods for the determination of protein-ligand dissociation constants. AB - This article is a review with 83 references of the application of NMR to the measurement of the dissociation constants of protein-ligand complexes. After briefly discussing some general concepts of molecular stability, the text turns to consider which NMR parameters are reporters of complex formation. The available data treatments required to translate observed NMR effects into quantitative measurements of the stability of the complex in the form of the dissociation constant (KD) are introduced. Linearisation methods and curve fitting methods are explained in detail and are illustrated with examples drawn from recent reports of protein-small molecule interactions. Throughout the text examples of the commonly observed NMR parameters , 1 / T1 and 1 / T2 are drawn from biological studies of 1H, 31P, 19F 15N (and other nuclei). The advantages of NMR diffusion experiments as a measure of KD are considered. Some less frequently used NMR approaches, some new ideas and some non-general methods are grouped together in a miscellaneous section. The major sources of errors in the determination of KD are identified. This allows recommendations for optimal experimental set up. Options for dealing with strong binding are reviewed. Finally, the implications of abstracting KD data from high throughput screening experiments are considered and several different approaches to generate this data are discussed. PMID- 12577992 TI - Contemporary issues in women's health. PMID- 12577991 TI - ALTEA: anomalous long term effects in astronauts. A probe on the influence of cosmic radiation and microgravity on the central nervous system during long flights. AB - The ALTEA project participates to the quest for increasing the safety of manned space flights. It addresses the problems related to possible functional damage to neural cells and circuits due to particle radiation in space environment. Specifically it aims at studying the functionality of the astronauts' Central Nervous Systems (CNS) during long space flights and relating it to the peculiar environments in space, with a particular focus on the particle flux impinging in the head. The project is a large international and multidisciplinary collaboration. Competences in particle physics, neurophysiology, psychophysiology, electronics, space environment, data analyses will work together to construct the fully integrated vision electrophysiology and particle analyser system which is the core device of the project: an helmet-shaped multi sensor device that will measure concurrently the dynamics of the functional status of the visual system and passage of each particle through the brain within a pre-determined energy window. ALTEA is scheduled to fly in the International Space Station in late 2002. One part of the multi-sensor device, one of the advanced silicon telescopes, will be launched in the ISS in early 2002 and serve as test for the final device and as discriminating dosimeter for the particle fluences within the ISS. PMID- 12577993 TI - History of medicine in the twentieth century: an overview. AB - The study of history is part of man's awareness of himself and the nature and place of his society in the world at large. The history of medicine is no exception. The past is always a guide to the present and the present sets the scene for the future. That is where lies the significance of the study of the history of medicine in the twentieth century. The medical achievements of this century ranks as one of the supreme epochs of human enterprise and endeavour. Medicine needed to march a par with the dynamic history of a changing scientific world. In fact, it did so. PMID- 12577994 TI - Imaging-guided extension of the time window: ready for application in experienced stroke centers? PMID- 12577995 TI - Emperor Akbar as a healer and his eminent physicians. AB - Akbar, however, was himself considered to be a healer to be healer by his miraculous powers. The court physician was one of the top-most officers of the court. Hakim Humam was such a physician who was one of the nine jewels (nauratnas) of Akbar. In 1581, Akbar curtailed the powers of Sadr (or Sadr-i Jahan, the chief Justice and Administrator of the Empire) and he divided the empire in six zones with a Sadr in charge of each. Out of these six Sadrs, five were his physicians who were Hakim Abul Fath, Shaikh Fayzi, Hakim Humam, Hakim Ali, Hakim Ain-ul-Mulk. Tobacco was introduced either in 1604 or in 1605. Hakim Abul Fath opposed its circulation but Akbar permitted it. It was Abul Fath who found out a remedy for counteracting the pernicious effects of tobacco on health. His opinion was that if the smoke of tobacco passed through water the harmful effects would be minimised. So hukka was invented by him. PMID- 12577996 TI - Clinical guidelines do serve their purpose. PMID- 12577997 TI - Nurses must be careful about medical roles. PMID- 12577998 TI - Nursing's practice and theory must be linked. PMID- 12577999 TI - Farming in space: environmental and biophysical concerns. AB - The colonization of space will depend on our ability to routinely provide for the metabolic needs (oxygen, water, and food) of a crew with minimal re-supply from Earth. On Earth, these functions are facilitated by the cultivation of plant crops, thus it is important to develop plant-based food production systems to sustain the presence of mankind in space. Farming practices on earth have evolved for thousands of years to meet both the demands of an ever-increasing population and the availability of scarce resources, and now these practices must adapt to accommodate the effects of global warming. Similar challenges are expected when earth-based agricultural practices are adapted for space-based agriculture. A key variable in space is gravity; planets (e.g. Mars, 1/3 g) and moons (e.g. Earth's moon, 1/6 g) differ from spacecraft orbiting the Earth (e.g. Space stations) or orbital transfer vehicles that are subject to microgravity. The movement of heat, water vapor, CO2 and O2 between plant surfaces and their environment is also affected by gravity. In microgravity, these processes may also be affected by reduced mass transport and thicker boundary layers around plant organs caused by the absence of buoyancy dependent convective transport. Future space farmers will have to adapt their practices to accommodate microgravity, high and low extremes in ambient temperatures, reduced atmospheric pressures, atmospheres containing high volatile organic carbon contents, and elevated to super-elevated CO2 concentrations. Farming in space must also be carried out within power-, volume-, and mass-limited life support systems and must share resources with manned crews. Improved lighting and sensor technologies will have to be developed and tested for use in space. These developments should also help make crop production in terrestrial controlled environments (plant growth chambers and greenhouses) more efficient and, therefore, make these alternative agricultural systems more economically feasible food production systems. PMID- 12578000 TI - Panduroga: a medico-historical study. AB - According to Ayurveda the word 'Pandu' denotes pale or yellowish white colour. Panduroga (anaemia) is a disease in which man becomes pallor due to deficiency of Rakta dhatu (blood) in the body. Rakta dhatu is mentioned among the Saptadhatus of the body. Historical importance of Panduroga and the comparative study regarding its Nidana-Samprapti, Lakshanas, Upadravas and Chikitsa etc. as found in Athavaveda, Mahabharata, Charaka Samhita, Sushruta Samhita, Chakradatta and Basava Rajeeyam etc. are being presented in this paper. PMID- 12578001 TI - ACOG practice bulletin: Shoulder dystocia. Number 40, November 2002. (Replaces practice pattern number 7, October 1997). PMID- 12578002 TI - Carbon balance in bioregenerative life support systems: some effects of system closure, waste management, and crop harvest index. AB - In Advanced Life Support (ALS) systems with bioregenerative components, plant photosynthesis would be used to produce O2 and food, while removing CO2. Much of the plant biomass would be inedible and hence must be considered in waste management. This waste could be oxidized (e.g., incinerated or aerobically digested) to resupply CO2 to the plants, but this would not be needed unless the system were highly closed with regard to food. For example, in a partially closed system where some of the food is grown and some is imported, CO2 from oxidized waste when combined with crew and microbial respiration could exceed the CO2 removal capability of the plants. Moreover, it would consume some O2 produced from photosynthesis that could have been used by the crew. For partially closed systems it would be more appropriate to store or find other uses for the inedible biomass and excess carbon, such as generating soils or growing woody plants (e.g., dwarf fruit trees). Regardless of system closure, high harvest crops (i.e., crops with a high edible to total biomass ratio) would increase food production per unit area and O2 yields for systems where waste biomass is oxidized to recycle CO2. Such interlinking effects between the plants and waste treatment strategies point out the importance of oxidizing only that amount of waste needed to optimize system performance. PMID- 12578003 TI - Patents, medicine, and the interests of patients: applying general principles to gene patenting. AB - New technologies and the translation of research into clinical medicine are essential to patient care. Those who develop useful drugs, diagnostic and screening tests, and medical technologies have the right to expect a fair return for their efforts and risks. Current interpretations of patent law (particularly regarding gene patenting), however, have the potential to impede both medical advances and patient care. Patenting policies must balance the open exchange and use of information with making the pursuit of such knowledge financially rewarding. The Committee on Ethics and the Committee on Genetics of the American College of Obstetricians and Gynecologists maintain that patents on medical or surgical procedures are ethically unacceptable. Physicians may obtain patents on surgical and diagnostic instruments that they have developed, but such instruments should be made available to others at a fair and reasonable cost. Patents for genes as "compositions of matter" enable patent holders to control future applications of the genes and should not be granted. Patents should be granted only for specified uses or applications of genes or sequences. If composition-of-matter patents on genes continue to be enforceable, such patents on genes with clinical applications should be subject to federal regulation and oversight to ensure availability on reasonable terms for research and clinical use. PMID- 12578004 TI - Screening of Madhumehaghna (anti diabetic) plants in Vrihattrayee. AB - The three texts written by Charak, Sushrut and Vagbhat are considered as Vrihattrayee because of their original contributions to the basic tenants of Ayurveda and innovative uses of plants and medicine. But despite the possibility of exploring efficacious for mulations from among these classics, not much attempts have been made in this direction, due to the fact that now most of them are not in vogue in practice by majority of Ayurvedic physicians. As such, a glossary of plants as described in those texts for the management of prameha including diabetes has been collected which would be useful for studying these drugs from different angles. PMID- 12578005 TI - Effects of air current speed on gas exchange in plant leaves and plant canopies. AB - To obtain basic data on adequate air circulation to enhance plant growth in a closed plant culture system in a controlled ecological life support system (CELSS), an investigation was made of the effects of the air current speed ranging from 0.01 to 1.0 m s-1 on photosynthesis and transpiration in sweetpotato leaves and photosynthesis in tomato seedlings canopies. The gas exchange rates in leaves and canopies were determined by using a chamber method with an infrared gas analyzer. The net photosynthetic rate and the transpiration rate increased significantly as the air current speeds increased from 0.01 to 0.2 m s-1. The transpiration rate increased gradually at air current speeds ranging from 0.2 to 1.0 m s-1 while the net photosynthetic rate was almost constant at air current speeds ranging from 0.5 to 1.0 m s-1. The increase in the net photosynthetic and transpiration rates were strongly dependent on decreased boundary-layer resistances against gas diffusion. The net photosynthetic rate of the plant canopy was doubled by an increased air current speed from 0.1 to 1.0 m s-1 above the plant canopy. The results demonstrate the importance of air movement around plants for enhancing the gas exchange in the leaf, especially in plant canopies in the CELSS. PMID- 12578006 TI - An update on the Kevorkian-Reding 93 physician-assisted deaths in Michigan: is Kevorkian a savior, serial-killer or suicidal martyr? AB - This report presents an update of the Kevorkian-Reding physician-assisted (or physician-aided) deaths to include the ninety-three publicly acknowledged cases as of November 25, 1998. These deaths are divided into ten distinct time phases. The following trends emerge. Over two-thirds of the decedents are women, the ratio of females to males varying widely with phase. The proportion of women seems to be the highest when Kevorkian is free to act as he wants and lowest when he seems to be acting under legal or political restraints. Based on autopsy results, only 29.0 percent of the cases are terminal, this percentage being higher among men (37.9%) than among women (25.4%). However, 66.7% of the decedents were disabled, no significant difference emerging between men and women. Further, five out of the six decedents showing no apparent anatomical sign of disease at autopsy were women. Over 80 percent of the physician-assisted deaths are cremated, approximately twice as high a proportion as that emerging for suicides in Michigan and four times as high as cremations occurring with regard to overall deaths. Finally, death by carbon monoxide decreases dramatically with time phase while the use of the contraption dubbed the "suicide machine" increases, suggesting an increasing routinization over time. Finally, during the ninth and tenth phases, Kevorkian's aims and his own suicidality emerge more clearly involving 1) harvesting organs and 2) threat of starving himself in prison if he is convicted. Phase 10 can be seen as an escalation from assisted death to overt euthanasia, repeating the same need for a demonstration (Thomas Youk) that was first exhibited in Phase I (Janet Adkins). PMID- 12578007 TI - Spaceflight hardware for conducting plant growth experiments in space: the early years 1960-2000. AB - The best strategy for supporting long-duration space missions is believed to be bioregenerative life support systems (BLSS). An integral part of a BLSS is a chamber supporting the growth of higher plants that would provide food, water, and atmosphere regeneration for the human crew. Such a chamber will have to be a complete plant growth system, capable of providing lighting, water, and nutrients to plants in microgravity. Other capabilities include temperature, humidity, and atmospheric gas composition controls. Many spaceflight experiments to date have utilized incomplete growth systems (typically having a hydration system but lacking lighting) to study tropic and metabolic changes in germinating seedlings and young plants. American, European, and Russian scientists have also developed a number of small complete plant growth systems for use in spaceflight research. Currently we are entering a new era of experimentation and hardware development as a result of long-term spaceflight opportunities available on the International Space Station. This is already impacting development of plant growth hardware. To take full advantage of these new opportunities and construct innovative systems, we must understand the results of past spaceflight experiments and the basic capabilities of the diverse plant growth systems that were used to conduct these experiments. The objective of this paper is to describe the most influential pieces of plant growth hardware that have been used for the purpose of conducting scientific experiments during the first 40 years of research. PMID- 12578008 TI - The 'principle', not 'doctrine' of double effect. PMID- 12578009 TI - Trust and betrayal in the medical marketplace. PMID- 12578010 TI - [Ubaldo Oppi: the 3 surgeons, 1926]. PMID- 12578011 TI - Dispensing death, desiring death: an exploration of medical roles and patient motivation during the period of legalized euthanasia in Australia. AB - A qualitative case study was conducted to explore the clinical decision making processes that underpinned the practice of euthanasia under the Rights of the Terminally Ill (ROTI) Act. The key informant for this research was Philip Nitschke, the general practitioner responsible for the legal cases of euthanasia. His information was supported by extensive document analysis based on the public texts created by patients in the form of letters and documentaries. Further collaborating sources were those texts generated by the media, rights groups, politicians, the coroner's cort, and the literature on euthanasia and assisted suicide. A key study finding was that the ROTI legislation did not adequately provide for the specific medical situation in the Northern Territory, Australia. The medical roles, as proscribed by the legislation, carried many inherent assumptions about the health care context and the availability of appropriately qualified medical staff committed to providing euthanasia. These assumptions translated into difficulties in establishing clinical practices for the provision of euthanasia. A further finding concerned the motivations of those who requested euthanasia. This article addresses the medical roles and the motivations of those seeking euthanasia. PMID- 12578012 TI - Hyperbaric oxygen prevents bacterial translocation in thermally injured rats. AB - This study was designed to evaluate the effects of hyperbaric oxygen (HBO2) on intestinal microflora and bacterial translocation (BT) caused by experimentally induced thermal injury in rats. Rats were separated into four groups, namely, HBO2 group, thermal injury (TI) group, TI + HBO2 group, and control group. All groups were further separated into short-term (2 days) and long-term (7 days) treatment or injury groups. Control group was neither exposed to thermal injury nor was given any treatment. Thirty percent second-degree thermal burn was induced on the dorsal body part of the rats in TI groups. In the HBO2 groups, rats received HBO2 treatment either without TI or following TI induction, for 2 and 7 days, respectively. Sampling from tissues and portal vein was performed on day 3 in the short-term groups and on day 8 in the long-term groups. Samples were cultured for identification of bacteria and colony counts. HBO2 treatment significantly reduced the colony counts of endogenous microflora in distal ileum of healthy rats (p < .05), while TI significantly increased the colony counts of endogenous microflora in distal ileum in short and long-term TI groups (p < .05). Presence of bacterial translocation was proven by bacterial isolation in mesenteric lymph nodes, liver, spleen and blood. Both short- and long-term HBO2 treatment following TI significantly reduced the colony counts of intestinal microflora (p < .05) and prevented bacterial translocation almost completely. It is concluded that thermal injury causes both bacterial overgrowth within intestinal lumen and bacterial translocation across the intestinal wall. HBO2 administration prevents both bacterial overgrowth and translocation. PMID- 12578013 TI - Ayurveda during Abbasid's period. AB - This is a historical paper which deals with a brief account of Abbasid's period. In this article the existence of Ayurveda in Arab countries, arrival of Ayurvedic physicians to Baghdad, their eminence, authenticity and literary additions in medical field has been studied and presented. PMID- 12578014 TI - References concerning birth control methods in Ancient Indian Classics: an exposition. AB - Reproduction is the basic phenomenon of Biological activity. Like other living beings man has a desire to reproduce. Unlike others he can control reproduction at his will. From early vedic periods belonging to several centuries prior to Christian era, we find references concerning methods of controlling the population over growth. Views of vedic sages, sexologists of ancient and medieval India regarding the birth control methods are expressed in this presentation. PMID- 12578015 TI - The bias against India in western literature on history of medicine: with special emphasis on public health. AB - The article describes a systematic bias against India in Western literature on history of medicine. While many authors have ignored the contributions of India in development of medicine altogether, the others have relegated India's role much behind other civilizations. Unnecessary and deliberate controversies on dating and origin of Ayurveda, primacy of Greek vs. Hindu Medicine and the origin of the practice of variolation have been elaborated by Western authors. Some medical historians, like Siegrist, have tried to give India its due place in the history of medicine. Suitable references of Indian authors have also been quoted to give a comparative and balanced picture. The need for settling this controversy has been emphasized. PMID- 12578016 TI - [Nursing care, rehabilitation, and nursing insurance for aged demented patients- the role of the attending physician in the system of care for the demented]. PMID- 12578017 TI - Obstetric causes for delivery of very-low-birth-weight babies at Tygerberg Hospital. PMID- 12578018 TI - A mass campaign too often? Results of a vaccination coverage survey in the Dikgale-Soekmekaar district. PMID- 12578019 TI - History of women in surgery: an overview. AB - The history of women as medical and surgical practitioners stretches back to at least 3500 B. C. Wall painting in tombs and temples of ancient Egypt show them performing surgical procedures and there is also evidence that women surgeons were practicing in ancient Sumeria, Babylon, Greece and Rome. During the middle ages as per formal decree issued by the church in the twelfth century monks were prohibited from "blood-letting" or performing operations. This effectively relegated wound treatment, bleeding and bone-setting to barbers and women. Women were particularly valued for their skill in obstetrics and midwifery. In this way an overview with regard to the history of women in surgery is presented here in chronological order. PMID- 12578020 TI - The Tughlaqs and medicine. AB - Muhammed Tughlaq established hospitals. A versatile scholar, Muhammed used to sit by the patients and watch the symptoms of extra ordinary diseases. His successor Firoz Tughlaq established a hospital where patients used to get free medical help and also food and drinks. This hospital was open to all. Though Firoz was known for his bigotry, we find no discrimination made between muslims and non-muslims in case of patients in this hospital. Firoz invented an eye-ointment in which, skin of black snake was used. It cured many kinds of eye diseases. This fact supports the Ayurvedic theory of and prescriptions for curing blindness with the ointment prepared with the flesh of cobras. Firoz himself was a good bone-setter. PMID- 12578021 TI - Bibliography of articles published in the Bulletin of the Indian Institute of History of Medicine (1997-1999). PMID- 12578022 TI - [The unfolding of the blood circulation doctrine: the correspondence between William Harvey and Caspar Hofmann in May 1636]. AB - The present article analyzes the letters exchanged by Caspar Hofmann and William Harvey in 1636 and the delineation of the main questions and answers on the blood circulation theory. The first part of the article presents concepts about blood circulation according to the time's physiology; the second part presents the changes introduced by Harvey; the third and last part presents the debate between Hofmann and Harvey. PMID- 12578024 TI - [Required clinical training for physicians and future training system]. PMID- 12578023 TI - [Postgraduate clinical training and university hospitals]. PMID- 12578025 TI - [Ideal postgraduate clinical training programs in internal medicine]. PMID- 12578026 TI - [Application of postgraduate clinical training program in an Anglo-American format]. PMID- 12578027 TI - [An attempt to construct a local network and suggestion for nationwide matching]. PMID- 12578028 TI - [Diagnosis and treatment of acute viral encephalitis]. PMID- 12578030 TI - Ergonomic automated anesthesia recordkeeper using a mobile touch screen with voice navigtion. AB - OBJECTIVE: To develop an ergonomically designed computerized recordkeeping tool for anesthesiologists that allows the clinician to maintain visual contact with the patient while performing recordkeeping. METHODS: To simplify the human interface software, we developed two general use software components. All purpose menu type 1 (APM1) was used for entering events using a tree structured menu. APM1 was designed to adapt to the limits of human memory, by using Miller's rule of 7 to guide the input process. APM1 can be considered to be a three-dimensional table list consisting of 7 vertical and 7 horizontal choices, which has further 5 tree-structured divergences. APM1 is also completely configurable by the user. All purpose menu 2 (APM2) was used to implement the system-initiated human interface where the system will prompt the user by voice for each entry. When users touch a key on APM1 and APM2, the system was designed to respond with a voice prompt. A touch-screen was also utilized and designed to fit the anesthesia machine. The screen is equipped with a small speaker for voice response and a microphone for voice recognition. The positions of the screen are adjustable supported by a long flexible limb (85 cm). RESULTS: After improving the design, systems were assembled for 10 operating rooms. Of the multiple features of the VOCAAR user interface, the following were well accepted by users and employed daily: touch-screen input, and voice response. The noncompulsory use rate was 87% during the initial 2 weeks, increased to 94% after 2 weeks and 100% after two months. The mean sound emission by voice response (n = 10, mean +/- SD) was 8.2 +/- 2.3 dB at the main anesthetist site (35 cm from the speaker mounted on the touch-screen), 2.2 +/- 1.3 dB at the staff site (1.5 m from the touch-screen), which was only audible for anesthesiologist but for surgeon. DISCUSSION: An EARK system was designed to allow the user to maintain visual contact with the patient while performing recordkeeping tasks. The combination of a mobile touch screen and voice response/recognition facilitated the design goals of the system. Although the system has enjoyed universal clinical acceptance, the voice functions remain too limited to satisfy the needs of a completely handsfree user interface. Enhancements to voice recognition technology will offer the potential for improved functionality. Additional research is also needed to better define the relationship between vigilance and visual contact with the patient. PMID- 12578029 TI - Selective versus non-selective neural stimulation in the monitoring of muscular relaxation during general anesthesia. AB - AIM: The depth of muscular relaxation during general anesthesia is monitored through the analysis of the contraction evoked by selective electrical stimulation of a peripheral nerve. The aim of this study was to compare the method of selective stimulation (SS) to a new method based on non-selective electrical stimulation (NSS) delivered over the muscle. METHOD: Electrical stimuli were delivered as train-of-four impulses to the ulnar nerve (SS) and to the ventral aspect of the contralateral forearm (NSS). The muscular responses of the adductor pollicis brevis (SS) and the forearm supinator longus (NSS) were studied at 30-60 s intervals with piezoelectric transducers before and after the administration of atracurium bolus doses of 0.5 mg/kg to patients under general anesthesia. SS and NSS evoked muscular responses were quantitized as percentages of the control response and compared with linear correlation and concordance analysis. RESULTS: Twenty patients were studied. Basal and post-atracurium muscular responses were similar for the SS and the NSS methods. Precision between SS and NSS was >85% and accuracy >92%. Concordance was: basal <15%, relaxation <5%, recovery <10%. CONCLUSION: NSS is equivalent to SS for muscular relaxation monitoring during general anesthesia. This has important implications to simplify muscular relaxation monitor design. PMID- 12578031 TI - New approaches for the detection and analysis of electroencephalographic burst suppression patterns in patients under sedation. AB - An automatic EEG pattern detection unit was developed and tested for the recognition of burst-suppression periods and for the separation of burst from suppression patterns. The median, standard deviation and the 95% edge frequency were computed from single channels of the EEG within a moving window and completed by the continuous computation of frequency band power via an adapted Hilbert resonance filter. These parameters were given to the inputs of two hierarchically arranged artificial neural networks (NNs). The output signals of NNs indicate the suppression and burst phases. The burst recognition was focused on the precise recognition of the burst onset. In subsequent processing steps the time course of percentages of burst patterns within their corresponding burst suppression-phases was calculated and the time locations of burst onsets can be used to trigger an averaging for a burst-related analysis. The data for our investigations were derived from the routine EEG derivations of 12 patients with various neurosurgical diseases. A group-related training of the NNs was realized. For the group-related trained NNs EEG data for 6 patients were used for training and the data of 6 other patients for testing the classification performance of the pattern recognition units. Additionally, the reliability of the detection algorithm was tested with data of two patients with convulsive state, resistant to treatment, and burst-suppression like pattern EEC. PMID- 12578032 TI - Artifact detection in the PO2 and PCO2 time series monitoring data from preterm infants. AB - BACKGROUND: Artifacts in clinical intensive care monitoring lead to false alarms and complicate later data analysis. Artifacts must be identified and processed to obtain clear information. In this paper, we present a method for detecting artifacts in PCO2 and PO2 physiological monitoring data from preterm infants. PATIENTS AND DATA: Monitored PO2 and PCO2 data (1 value per minute) from 10 preterm infants requiring intensive care were used for these experiments. A domain expert was used to review and confirm the detected artifact. METHODS: Three different classes of artifact detectors (i.e., limit-based detectors, deviation-based detectors, and correlation-based detectors) were designed and used. Each identified artifacts from a different perspective. Integrating the individual detectors, we developed a parametric artifact detector, called ArtiDetect. By an exhaustive search in the space of ArtiDetect instances, we successfully discovered an optimal instance, denoted as ArtiDetector. RESULTS: The sensitivity and specificity of ArtiDetector for PO2 artifacts is 95.0% (SD = 4.5%) and 94.2% (SD = 4.5%), respectively. The sensitivity and specificity of ArtiDetector for PCO2 artifacts is 97.2% (SD = 3.6%) and 94.1% (SD = 4.2%), respectively. Moreover, 97.0% and 98.0% of the artifactual episodes in the PO2 and PCO2 channels respectively are confirmed by ArtiDetector. CONCLUSIONS: Based on the judgement of the expert, our detection method detects most PO2 and PCO2 artifacts and artifactual episodes in the 10 randomly selected preterm infants. The method makes little use of domain knowledge, and can be easily extended to detect artifacts in other monitoring channels. PMID- 12578033 TI - Validation of an ultrasound scanner for determing urinary volumes in surgical patients and volunteers. AB - OBJECTIVE: As bladder distension related to anaesthesia puts patients at risk for permanent dysfunction, peri-operative determination of bladder volume is of great importance. The aim of this study is to validate an ultrasonic imaging device for determing bladder urine volume. METHOD: To evaluate a broad volume range, ultrasonically scanned volumes were compared to true urinary volumes both in surgical patients and in volunteers. After institutional approval and informed consent 60 healthy volunteers were asked not to void for as long as possible. After ultrasound measurements (BladderScan BVI 2500, Diagnostic Ultrasound, Redmond WA, U.S.A.) they voided and true urinary volumes were measured. Fifty surgical patients scheduled for procedures requiring urinary catheterisation were studied. Pre- and post-induction of anaesthesia ultrasound measurements were recorded, followed by urinary catheterisation and measurement of true urinary volume. Urine volumes were compared using Student t-tests and Wilcoxon Rank Tests (p < 0.05). For validation linear regression was used together with Bland-Altman analyses. RESULTS: Ultrasonic scanning underestimated the true urine volume by about 7% over the whole volume range (17 ml to 970 ml). Underestimation was larger in females than in males (p < 0.02). R2 values for correlation of measured and scanned urinary volumes ranged between 0.92 and 0.95. Bland and Altman analyses showed a bias of 31 ml in volunteers and of 19 ml in patients and a precision of 110 ml and 80 ml, respectively. CONCLUSIONS: The ultrasonic imaging device can be used peri-operatively to establish bladder volume, taking into account the 7% underestimation of the bladder volume. PMID- 12578034 TI - Partial CO2 rebreathing cardiac output--operating principles of the NICO system. AB - The partial rebreathing method of cardiac output estimation is reviewed with a particular focus on its application for continuous monitoring, rebreathing and implementations and from both a historical and technical perspective. The assumptions of the method are discussed as well as the various implementations. The NICO monitor and rebreathing valve are described from a functional view. The clinical data including (a) comparisons between bolus thermodilution and continuous thermodilution in patients in the OR setting, (b) comparisons to continuous thermodilution with both the Baxter and Abbott continuous cardiac output devices and (c) comparison between different means of shunt correction are presented. Compared to conventional cardiac output methods, the partial CO2 rebreathing technique is non-invasive, can easily be automated and can provide real-time and continuous cardiac output monitoring. Taking advantage of modern sophisticated sensor and signal processing technology and integrating multiple monitored physiological variables the NICO monitor is the first commercially available cardiac output system making use of the partial rebreathing of CO2. PMID- 12578035 TI - Microstream capnograpy technology: a new approach to an old problem. AB - Significant technical limitations inherent in black-body infrared technology used in conventional sidestream and mainstream capnography have hindered the acceptance and growth of capnography as a monitoring tool outside the operating room environment. We describe a new technology (Microstream) for CO2 monitoring, based on molecular correlation spectroscopy, which results in a highly efficient and selective emission of a spectrum of discrete wavelengths exactly matching those for CO2 absorption. The CO2 specific emissions allow for an extremely small sample cell (15 microl), which in turn, permits the use of a very low sample flow rate (50 ml/min) without compromising waveform integrity or end-tidal CO2 accuracy. Design and technology features of the CO2 emission source, sample cell, and breath sampling circuits are described. PMID- 12578036 TI - The challenges of human engineering research. PMID- 12578037 TI - Information services and the Internet. PMID- 12578038 TI - Validation of the Natus CO-Stat End Tidal Breath Analyzer in children and adults. AB - OBJECTIVE: The performance of a point-of-care, noninvasive end tidal breath carbon monoxide analyzer (CO-Stat End Tidal Breath Analyzer, Natus Medical Inc.) that also reports end tidal carbon dioxide (ETCO2) and respiratory rate (RR), was compared to established, marketed (predicate) devices in children (n = 39) and adults (n = 48) who are normal or at-risk of elevated CO excretion. METHODS: Concentrations of end tidal breath CO (ETCO), room air CO, ETCO corrected for inhaled CO (ETCOc), ETCO2, and RR were measured with the CO-Stat analyzer and the data compared to those obtained from the same subjects using the Vitalograph BreathCO monitor (Vitalograph, Inc.) for ETCOc and the Pryon CO2 monitor (SC210 and SC300, Pryon Corp) for ETCO2 and RR. Adults and children were studied at three medical centers. The data were analyzed by paired t-tests and linear regression. Bias and imprecision between the CO-Stat analyzer and the predicate devices was calculated by the method of Bland and Altman. RESULTS: Paired t tests, performed on the three parameters measured with the CO-Stat analyzer and predicate devices showed that only the ETCOc values in the adults and the ETCO2 values in the children were significantly different (lower, p < or = 0.0001, and higher, p < or = 0.0001, respectively). The mean bias and imprecision of the CO Stat analyzer for adult ETCOc and children ETCO2 measurements were -0.9 +/- 1.2 ppm and 0.4 +/- 0.6%, respectively. Linear regression analysis for the ETCOc results in children and adults had a high degree of correlation (r = 0.91 and 0.98, respectively). CONCLUSIONS: We conclude that in a clinical environment the Natus CO-Stat End Tidal Breath Analyzer performs at least as well as predicate devices for the measurements of ETCOc, ETCO2, and RR. PMID- 12578039 TI - Hemodilution does not alter the aortic-to-femoral arterial pressure difference in dogs. AB - INTRODUCTION: Distal arterial pressure normally differs from aortic pressure. This difference is modified by changes of vascular resistance. Hemodilution, due to decreased viscosity, decreases vascular resistance. Therefore, the difference between aortic and distal arterial pressures could be altered as well. We investigated whether acute hemodilution affected this difference in dogs. METHODS: Eleven mongrel dogs weighing 16.6 +/- 4.4 kg were anesthetized with pentobarbital and sufentanyl and mechanically ventilated. Arterial presssure was recorded using Millar catheter-tipped pressure transducers in the proximal aorta and in the distal femoral artery. An electromagnetic flowmeter probe was placed around the aorta. Effective downstream pressure was estimated by extrapolation of exponential arterial pressure decay during 3-second occlusion of the proximal aorta. Hemodilution was effected by removal of 30 ml/kg of blood and replacement with 60 ml/kg of warmed saline. In addition, the effects of 1 microg/kg phenylephrine and 4 microg/kg of sodium nitroprusside were measured before and after hemodilution. RESULTS: Hemodilution decreased hematocrit from 39 +/- 11.2% to 25.6 +/- 4.95%. Systolic and mean pressures were unchanged but aortic diastolic pressure decreased significantly, from 86 +/- 17 to 79 +/- 15 mmHg (p < 0.005). Peak systolic pressure was 13.5 +/- 7.2 mmHg higher in the femoral artery than in the aorta before, and 16 +/- 8.7 mmHg after, hemodilution (p > 0.05). Nitroprusside decreased the femoral to aortic peak systolic pressure difference from 14.3 +/- 6.3 to 7.7 +/- 15.3 mmHg, p = 0.05 before hemodilution and from 14.3 +/- 8.8 to 2.5 +/- 11 mmHg, p < 0.005 afterwards. Hemodilution significantly decreased the effective downstream pressure, from 44 +/- 9 to 36 +/- 6.8 mmHg in the aorta (p < 0.05), and from 51 +/- 2 to 37 +/- 3.1 mmHg in the distal femoral artery (p < 0.05). CONCLUSION: Acute hemodilution did not alter the aortic-to distal arterial pressure difference in dogs. PMID- 12578040 TI - Response time of the Opti-Q continuous cardiac output pulmonary artery catheter in the urgent mode to a step change in cardiac output. AB - OBJECTIVES: This study was conducted to determine the response time of the Opti-Q continuous cardiac output (CCO) device to a step change in cardiac. DESIGN: Prospective study. SETTING: University hospital animal lab. MODEL: Female sheep. INTERVENTIONS: In ten animals, cardiac output was altered suddenly by opening and closing a peripheral arteriovenous shunt to test the response time of the CCO system. MEASUREMENTS AND MAIN RESULTS: Cardiac output was measured continuously by thermodilution and ultrasonic techniques while an arteriovenous shunt was opened and closed. A total of 53 dynamic observations were made (5-6 per animal). The mean response time of the continuous cardiac output device was 86 seconds and was unaffected by the magnitude or direction of the change in cardiac output. It was also unaffected by the animal's weight. CCO values were not statistically different from standard thermodilution measurement (p = 0.895). Shunt flow ranged from 430 to 1730 ml/min and averaged 812 ml/min. The mean CCO with the shunt closed was 4.62 L/min. There was 1.5 to 2 minutes under or overshoot in cardiac output in 11% of the measurements. CONCLUSIONS: Continuous cardiac output measurement was as accurate as those made by standard bolus thermodilution. The average response time to acute changes in cardiac output was approximately 1.5 minutes or ten times faster than previously reported systems. Response time is independent of animal mass, shunt volume and the direction of cardiac output perturbations. PMID- 12578041 TI - A comparison of pulse oximetry and near infrared spectroscopy (NIRS) in the detection of hypoxaemia occurring with pauses in nasal airflow in neonates. AB - OBJECTIVE: The aim of this study was to compare the ability of NIRS and pulse oximetry to detect changes in cerebral oxygenation occurring in response to a pause in nasal airflow (PNA). METHODS: Twenty-one recordings of cerebral oxygenation index by NIRS together with oxyhemoglobin saturation by pulse oximetry were measured on 17 preterm infants with a history of apnoea. Photoplethysmography was used to confirm the accuracy of the pulse oximetry data. PNA events were defined as pauses of greater than 4 seconds in a thermistor trace measuring nasal air flow. RESULTS: Baseline variability in oxygenation index (Hbdiff) was found to be from -0.12 to +0.13 micromol 100 g brain(-1). A fall in Hbdiff or SpO2 was defined as a decrease of greater magnitude than 2 standard deviations from the baseline, i.e., -0.12 micromol 100 g brain(-1) and 3% respectively. In 68% of 468 PNA events a fall in oxyhemoglobin saturation (SpO2) was detected and in 56% a fall in Hbdiff was detected. In 20% of events there was no fall in cerebral oxygenation despite a fall in SpO2. In 8% of PNA episodes we recorded a fall in cerebral oxygenation but no fall in SpO2. When a fall in cerebral oxygenation was recorded, the fall was greater when the event was also associated with a fall in SpO2 (median (interquartile range (IQR)) 0.32 (0.21 0.69) vs. 0.25 (0.16-0.43) micromol 100 g brain(-1), p < 0.05). When all the PNA episodes were reviewed no close correlation was shown between the magnitude of change in cerebral oxygenation and the change in SpO2 for small changes in both indices. However, large falls (>1.5 micromol 100 g brain(-1)) in cerebral oxygenation were closely associated with large changes in SpO2. CONCLUSIONS: We conclude that both techniques are sensitive to changes in oxygenation during PNA. Small changes in cerebral Hbdiff and arterial SpO2 do not always correlate for physiological reasons. A change in Hbdiff of >0.3 micromol 100 g brain(-1) is likely to be physiologically significant and is associated with a change in SpO2 of 12%. PMID- 12578042 TI - A comparison of the Internet and the standard textbook in preparing for the professional anaesthetic examination. AB - The Internet is increasingly being recognised as a source of information in different fields, and medicine is no exception. We investigated the use of the Internet as an aid in preparing for the professional medical examinations. The Internet was compared to a standard textbook in answering a randomly selected past examination paper. Thirty-eight web-sites on the Internet were involved, giving information to all the examination questions. In contrast, the textbook provided adequate information in only 73.3% of the questions. The time required to search for information on the Internet was more than that required for a single textbook. (approximately 2 hours per question compared to 30 minutes per question respectively). However, with the rapid development of computer technology and the cyberspace, the Internet may prove to be a viable alternative or a good supplement to the standard textbook when trainee anaesthetists are preparing for their professional medical examinations. PMID- 12578043 TI - A method for overcoming the ceiling effect of bounded pain scales. AB - OBJECTIVE: The Verbal Numerical Scale (VNS) for rating pain is bounded between 0 (= no pain) and 10 (= worst pain imaginable). We hypothesized that the limitations inherent to this boundary when rating extremely painful stimuli may be identified by integrating the VNS with an unbounded score such as magnitude estimation of relative change. METHODS: Volunteers received stimuli of increasing current via cutaneous electrodes until they rated >5 on the VNS scale. This stimulus, termed S, was arbitrarily assigned a magnitude estimate of 100%. Then, stimuli of varying currents were delivered; two were 10 mA and 20 mA higher than S (S(+10) and S(+20)), two were 1/2 of the current for the S stimulus (S(1/2)), and one was at the original current (Srepeat). The pain elicited by each stimulus was scored in proportion to the S stimulus. The extrapolated VNS score (VNSext) was determined by multiplying this magnitude estimate (%) by the VNS score for S. MAIN RESULTS: Seventy percent of the stimuli with higher intensity than S generated a VNSext score above 10. The mean magnitude estimations for S(+10) and S(+20) were 186% and 242%: they generated mean (median) VNSext values of 12.4 and 16.2, respectively (p = 0.019 for the difference between them by Wilcoxon signed rank test). CONCLUSIONS: The combined use of VNS and magnitude estimation confirmed that the ceiling of the bounded pain scale may significantly limit a patient's ability to describe a new pain stimulus. VNSext may provide a means of overcoming this limitation. PMID- 12578044 TI - Monitoring of heart and respiratory rates in newborn infants using a new photoplethysmographic technique. AB - OBJECTIVE: A new photoplethysmographic (PPG) device for respiratory and heart rate monitoring has been evaluated in the neonatal care units at the University Children's Hospital of Uppsala, Sweden. The purpose of this study was to compare this new device with more established techniques, i.e., transthoracic impedance plethysmography (TTI) for monitoring of respiratory rate and ECG for heart rate monitoring. METHODS: Data were acquired continuously for 8-hours in each of 6 neonates. The signals were analysed for periods of 30 seconds, in which the heart and respiratory signals from the PPG device were compared with the ECG and the impedance plethysmogram. RESULTS: The ECG recordings were of high quality in 77% of the analysed periods. In these periods, excluding periods (6%) disturbed by offset-adjustement of the PPG signal, the PPG heart signal included 1.1% (+/- 0.7% SD) false negative beats and 0.9% (+/- 0.6%) false positive beats. In periods with an impedance signal of high quality (29% of total time), the part of the PPG signal synchronous with respiration included 2.7% (+/- 1.1%) false negative breaths and 1.5% (+/- 0.4%) false positive breaths. Here, 2% of the periods were discarded because of offset-adjustment. From the periods of low signal quality, two other conclusions were drawn: 1) The impedance signal contains more power in the respiratory range than the corresponding PPG respiratory signal. 2) The breaths are easier to identify in the PPG respiratory signal than in the impedance signal (subjective measure). CONCLUSIONS: Electrode and motion artefacts seem to disturb the ECG signals and, particularly, the impedance signals. During periods of high quality ECG and impedance signals, the new optical device produces signals of equal quality to these traditional methods, and is in some cases even better. The new device is non-invasive and has a small optical probe. These factors indicate further advantages of the photoplethysmographic method. PMID- 12578045 TI - Ergonomic evaluation of an ecological interface and a profilogram display for hemodynamic monitoring. AB - OBJECTIVE: Comprehensive monitoring of the patient state and subsequent decision making is an essential part of the task of an anaesthetist. The physicians' decision making process is based upon a concept of partly abstract physiologic parameters such as depth of anaesthesia or contractility. This concept is derived from the measured parameters given on todays' trend displays in addition to context information available for the anaesthetist. We investigated two alternative approaches of display design for hemodynamic monitoring: 1) integrated displays based on ecological interface design, and 2) profilogram displays based on intelligent alarms. METHOD: To evaluate differences in decision making, the two displays and a trend display were compared in an experimental set up with computer simulated vital parameter curves. From a start state with random parameter deviations from the ideal state, subjects had to achieve the ideal circulatory performance as fast as possible by manipulating vasomotor tone, heart rate, blood volume and contractility. To analyse subjects' decision making process, eye-tracking, event-logging, and the method of think aloud protocols were used. Twenty anaesthesiologists performed 113 experiments (approximately 2 with each display). RESULTS: The anaesthetists failed to achieve the task in 37% using the trend display, in 19% using the profilogram display, and in 13% using the ecological interface. Hence, a safer task solution was possible with the ecological interface and the profilogram display but at the expense of various performance parameters such as higher trial time, more interactions with the simulated system, and more frequent eye movements. In contrast to the trend display and the profilogram display, where anaesthetists were mainly focussed on controlling the left atrial pressure, such an behaviour was less observed with the ecological interface. CONCLUSION: Our results have shown that subjects came to more effective solutions with the traditional trend display. The main reason for this result may be their years of experience with this kind of display type. Regarding safe and goal-intended decision finding, the results are encouraging for further experiments with redesigned ecological displays. But these displays ought to have smoother changes with respect to the traditional trend displays. Furthermore, new experiments have to be performed under real or fairly real (e.g. together with an anaesthesia simulator) conditions to underline the positive results for ecological interfaces. PMID- 12578046 TI - Autoregulation in a simulator-based educational model of intracranial physiology. AB - OBJECTIVE: To implement a realistic autoregulation mechanism to enhance an existing educational brain model that displays in real-time the cerebral metabolic rate (CMRO2), cerebral blood flow (CBF), cerebral blood volume (CBV), intracranial pressure (ICP), and cerebral perfusion pressure (CPP). METHODS: A dynamic cerebrovascular resistance (CVR) feedback loop adjusts automatically to maintain CBF within a range of the CPP and defines autoregulation. The model obtains physiologic parameters from a full-scale patient simulator. We assumed that oxygen demand and arterial partial pressure of carbon dioxide (CO2 responsivity) are the two major factors involved in determining CBF. In addition, our brain model increases oxygen extraction up to 70% once CBF becomes insufficient to support CMRO2. The model was validated against data from the literature. RESULTS: The model's response varied less than 9% from the literature data. Similarly, based on correlation coefficients between the brain model and experimental data, a good fit was obtained for curves describing the relationship between CBF and PaCO2 at a mean arterial blood pressure of 150 mm Hg (R2 = 0.92) and 100 mm Hg (R2 = 0.70). DISCUSSION: The autoregulated brain model, with incorporated CO2 responsivity and a variable oxygen extraction, automatically produces changes in CVR, CBF, CBV, and ICP consistent with literature reports, when run concurrently with a METI full-scale patient simulator (Medical Education Technologies, Inc., Sarasota, Florida). Once the model is enhanced to include herniation, vasospasm, and drug effects, its utility will be expanded beyond demonstrating only basic neuroanesthesia concepts. PMID- 12578047 TI - The relationship between modified pulse wave transit time and cardiovascular changes in isoflurane anesthetized dogs. AB - OBJECTIVE: To clarify the relationship between blood pressure and pulse wave transit time at the peripheral artery from the R wave of the electrocardiogram (m PWTT), the effects of cardiovascular interventions on this relationship was evaluated. METHODS: Ten mongrel dogs were anesthetized by isoflurane inhalation, and catheter tip pressure transducers were inserted into the ascending aorta and at the bifurcation of abdominal aorta to measure central and peripheral pulse wave arrival. Pulse wave arrival at the ascending aorta from the R wave represents pre-ejection period (PEP) and pulse wave arrival between the ascending aorta and bifurcation of aorta represents pulse wave transit time (PWTT), thus m PWTT = PEP + PWTT. Hypertension was induced by the continuous infusion of dobutamine and phenylephrine, and hypotension was induced by deepening isoflurane anesthesia, acute blood loss and nitroglycerine infusion. The relationship between timing components (PWTT, PEP, and m-PWTT) and blood pressure was recorded and analyzed by using the least squares method. RESULTS: The relationship between timing components (PWTT, PEP and, m-PWTT) and blood pressure was significant and highly correlated. When the change in blood pressure was due to the myocardial contractility, such as after dobutamine infusion, the relationship between all timing components and blood pressure was consistent and negative. However, when the change in blood pressure was due to the vasoactive agents, such as phenylephrine, the relationship between timing components and blood pressure was dependent on the reflex change in PEP. CONCLUSIONS: Change in m-PWTT is a good parameter to predict blood pressure changes, although the absolute blood pressure value cannot be obtained. PMID- 12578048 TI - Patient's perceptions of an anesthesia preoperative computerized patient interview. AB - Our desire to elicit a more complete medical history from our patients led to the implementation of a preoperative computerized interview. We previously demonstrated the effectiveness of the interview by computing its mean completion time for the overall patient population (n = 120), and further examined the effects of age, gender, and educational level. In this study, we investigated patient perception of the interview itself. Before and after taking the computer interview, we asked the patients to complete a paper and pencil questionnaire comprised of sixteen questions, expressing their feelings toward the computer interview. Responses elicited prior to taking the computer interview were compared with those obtained afterward. The Stuart-Maxwell test was used to determine statistically significant differences in answers before and after the interview. Initial questionnaire responses reflected a positive attitude toward computer usage which became even stronger after the interview. The only negative responses elicited were really more "doctor positive" than "computer negative." We conclude that patients looked favorably upon participating in a computerized medical interview provided that physician-patient contact is maintained. PMID- 12578049 TI - Inhaled nitric oxide fraction is influenced by both the site and the mode of administration. AB - OBJECTIVE: Inhaled nitric oxide (NO) can be delivered continuously or sequentially (= during inspiration) at different locations of the ventilation circuit. We have tested the influence of locations, modes of NO administration and the ratio of the inspiratory time over the respiratory cycle time (I/I + E ratio) on the accuracy of NO fractions, delivered by 2 devices: Opti-NO and Flowmeter. METHODS: We used a simplified lung model consisting of a ventilation circuit with a Y piece, a tracheal tube, a 150 ml dead-space volume and a 5 liter balloon. Three fractions (3, 6, 9 ppm) were administered continuously or sequentially, in controlled volume, in 4 different sites on the inspiratory branch above the Y piece: i) just after the water trap, ii) just before the Y piece; below the Y piece: iii) just after the Y piece, iv) into the endotracheal tube. In addition, different I/I + E ratios (25, 33, 50, 80%) were studied. The delivered NO fractions were measured in the balloon by chemiluminescence (CLD 700, Ecophysics). A linear regression analysis was used to test the relationship between administered and measured NO fractions for the 3 fractions (3, 6 and 9 ppm) in sequential and continuous modes. Intercept values were compared to zero and slopes to the identity line. RESULTS: When NO was administered in the continuous mode upstream the Y piece, NO fractions measured in the balloon corresponded to the administered fractions. In contrast, below the Y piece, the measured NO fractions were significantly lower than the administered NO fractions. In the sequential mode, above and below the Y piece, the delivered NO fractions were within the manufacturer's range. CONCLUSIONS: For the continuous NO delivery, locations above the Y piece are mandatory. However, locations below the Y piece imposes a sequential system, which can also be used for the sites located above the Y piece. PMID- 12578050 TI - The influence of left lateral position on cardiac output changes after head up tilt measured by impedance cardiography. AB - BACKGROUND: The value of the impedance cardiography (IC) method for measuring cardiac output (CO) with the subject in the left lateral position has not yet been established. We compared the CO after a 30 degrees head-up tilt with the subjects in the supine and left lateral position. METHODS: Thirty healthy young volunteers were placed in the supine horizontal position. CO, stroke volume (SV) and heart rate (HR) were measured for five minutes using impedance cardiography (NCOMM3, BoMed Medical Manufacturing, Location). Then a 30 degrees head-up tilt was done and the measurements were continued for an additional five minutes. After that the subjects were placed in the left lateral horizontal position and the measuring procedure was repeated. RESULTS: After the tilt, SV and cardiac index (CI) decreased and HR increased statistically significantly in the supine and in the left lateral position. SV and CI also decreased statistically significantly but HR did not change after the shift from the supine to the left lateral position. Although CI was smaller in the left lateral position, the time course of CI change after the tilt was identical in the supine and in the left lateral position. CONCLUSION: When using the IC method of SV measurement, the absolute value of the CI changes when moving from the supine to lateral positions. It is not clear whether this change is physiologic or an artifact of the measurement technique. However, changes in CI in response to a 30 degree head up tilt are the same in either position. We conclude that changes in CI can be measured with the IC method in the lateral position. PMID- 12578051 TI - A comparison of lithium dilution cardiac output measurements made using central and antecubital venous injection of lithium chloride. AB - OBJECTIVE: We have previously described an indicator dilution technique of measuring cardiac output in which lithium chloride is injected as a bolus via a central venous catheter and cardiac output derived from the arterial lithium dilution curve recorded from a lithium-selective electrode, which we have developed for this purpose. It would be an advantage if the lithium could be injected via the basilic vein (in the antecubital fossa) in those patients who do not need central venous catheterisation for other reasons. We have therefore compared cardiac output measurements made using these two routes of lithium chloride administration. METHODS: Lithium dilution cardiac output was measured 10 times in each of 10 patients, injecting the lithium chloride alternately via the basilic or central venous catheter. RESULTS: The mean difference was 0.8 +/- 5.2% (SD) (range -8.5 to +7.0%) over a range of cardiac output of 4.5-13 l/min. CONCLUSIONS: Injection of lithium chloride via the basilic vein in the antecubital fossa allows accurate lithium dilution cardiac output measurements to be made in patients who do not have central venous catheters in place. PMID- 12578052 TI - Drug levels: therapeutic and toxic serum/plasma concentrations of common drugs. PMID- 12578053 TI - Work analysis--a first step to understanding modes of system failure and developing rational remedies. PMID- 12578054 TI - Vigilance, boredom, and sleepiness. PMID- 12578055 TI - Computerized monitoring of physical activity and sleep in postoperative abdominal surgery patients. AB - OBJECTIVE: Assessment of early postoperative activity is important in the documentation of improvements of peri-operative care. This study was designed to validate computerized activity-based monitoring of physical activity and sleep (actigraphy) in patients after abdominal surgery. METHODS: The study included twelve hospitalized patients after major abdominal surgery studied on day 2 to 4 after operation and twelve unhospitalized healthy volunteers. Measurements were performed for 24 consecutive hours. The actigraphy measurements were compared with self-reported activity- and sleep registration. The actigraphy output was obtained by the zero-crossing mode (ZCM) and time-above-threshold mode (TATM). RESULTS: The overall results showed comparable mean agreement between actigraphy and self-reported activity registrations for patients of 80% (SD 12%) and volunteers of 84% (SD 6%) (p = 0.15). In both study groups, all correlation coefficients between actigraphy measurements and self-reported activity data were statistically significant (r, values for patients 0.4 to 0.8 and volunteers 0.6 to 0.9). A higher mean agreement between automated actigraphy sleep detection and self-reported sleep registration were found in the volunteers (85%, SD 15%) compared with the patients (77%, SD 11%) (p < 0.05). The mean activity value awake was higher in the volunteers than in the patients (p < 0.05). CONCLUSION: Computerized activity monitoring by actigraphy is a reliable and easy method for monitoring physical activity and sleep-wake cycles after major abdominal surgery. PMID- 12578056 TI - A single computer-controlled mechanical insufflation allows determination of the pressure-volume relationship of the respiratory system. AB - OBJECTIVE: To evaluate and further develop a method for determination and mathematical characterisation of the elastic pressure-volume (Pel-V) relationship in mechanically ventilated human subjects during one single modified insufflation with simultaneous determination of resistance of the respiratory system. SUBJECTS: Eight adult non-smoking human subjects without heart, lung, or thoracic cage disease scheduled for non-thoracic surgery. The study was performed in anaesthetised and muscle-relaxed subjects. MEASUREMENTS AND MAIN RESULTS: The Pel V curve was determined with a computer-controlled Servo Ventilator 900C during a modified insufflation with either constant or sinusoidally varying flow. Pressure and flow were measured with the built-in sensors of the ventilator. Tracheal pressure (Ptr) was calculated by subtracting the pressure drop over the tracheal tube. The elastic recoil pressure in the peripheral lung, Pel, was obtained from the calculated Ptr by subtracting the pressure drop over the airways. Ptr was also directly measured through a catheter. The calculated Ptr gave similar results as the directly measured Ptr, thus indicating the reliability of the signal originating from the ventilator sensor for computation of downstream pressures. The inflection points of the sigmoidal Pel-V curve and the compliance of the linear segment were determined with high reproducibility. CONCLUSIONS: Using one single modified insufflation allows a fast and accurate determination of respiratory mechanics. The Pel-V curves were determined with high reproducibility and were adequately described by a three-segment model of the curve incorporating a linear segment between two asymmetrical non-linear segments. PMID- 12578057 TI - A simple strategy for faster induction and more cost-effective use of anesthetic vapor. AB - INTRODUCTION: Inducing general anesthesia often involves mask ventilation using high fresh gas flow (FGF) to administer anesthetic vapor prior to endotracheal intubation. A common practice is to turn the vaporizer off when the mask is removed from the patient's face to avoid room contamination (VAPOff). An alternative approach is to leave the vaporizer on and turn the FGF to minimum to reduce the amount of vapor laden gas that can enter the room (FGFOff). The objective of this study is to compare the relative induction times and vapor costs associated with each induction strategy. METHODS: Each induction method was simulated using Gasman (MedMan Simulations, Chestnut Hill, MA) for Windows assuming a 70 kg patient. To simulate a period of mask ventilation with anesthetic vapor prior to intubation, the FGF was set to 6 l/min and the isoflurane vapor concentration to 1.2% (1 MAC) for three minutes with an alveolar ventilation of 5 l/min and cardiac output of 5 l/min. For the first simulation of the intubation period (FGFOff), FGF was turned to 150 ml/min, minute ventilation was set to zero and the vaporizer setting unchanged for one minute. Initial settings were then restored and the rate of change of anesthetic vapor concentration in the circuit (Vckt) and alveolus (Valv) followed for 10 minutes along with the cost of delivered vapor (V(S)). For the second simulation (VAPOff), after the initial three minutes of vapor delivery, the vaporizer was set to zero, minute ventilation was set to zero and the FGF left unchanged for one minute. The initial settings were then restored and Vckt, Valv and V(S) followed for ten minutes. The cost calculation was based upon a 100 ml bottle of Isoflurane at $72/bottle. Actual gas flow was measured at the y-piece of a circle system for fresh gas flows from 0.15 to 6 l/min. RESULTS: At the end of the simulated intubation period (minute 4), Vckt was unchanged with the FGFOff method whereas it had fallen by more than half with the VAPOff method. Using VAPOff, it took until the 6-minute mark for Vckt to return to the same concentration that existed prior to intubation at minute three. Throughout the 10 minute simulated induction, Valv using FGFOff exceeded Valv using VAPOff although the difference became small at the end of the period. V(S) was essentially identical at all time points. No flow into the room was measured at the minimum fresh gas flow whereas higher fresh gas flows resulted in a significant portion of the fresh gas flowing into the, room. CONCLUSIONS: The strategy of turning the FGF to minimum and leaving the vaporizer on during intubation does not contaminate the room and speeds induction by fostering a greater alveolar concentration than the VAPOff method. Cost savings are derived using FGFOff since a higher alveolar concentration is achieved at the same vapor cost. Additional cost savings are demonstrated since a low flow technique is possible immediately after intubation when using FGFOff. The practice of turning off the vaporizer during endotracheal intubation while FGF remains high should be abandoned. PMID- 12578058 TI - Clinical investigation of a new combined pulse oximetry and carbon dioxide tension sensor in adult anaesthesia. AB - OBJECTIVE: To test the accuracy of a new combined oxygen saturation and cutaneous carbon dioxide tension (SPO2-PCO2) sensor in a routine adult clinical environment. This probe provides a non-invasive and continuous monitoring of the arterial oxyhaemoglobin saturation, arterial carbon dioxide tension and pulse rate at the ear lobe. The sensor is intended to measure both relevant respiration/ventilation parameters in one single probe. METHODS: Ten adult patients were consecutively studied during general anaesthesia. During the first 5 min after sensor placement at the ear lobe, arterial blood samples were drawn each minute. Carbon dioxide tension and oxygen saturation measurements were obtained simultaneously at 1-min intervals. After this period, patients were hyper-, normo- and hypoventilated. After 15 min at each setting, the simultaneously obtained cutaneous and arterial carbon dioxide tension values were compared. RESULTS: A total of 80 comparisons between ear lobe SpO2-PCO2 measurement, finger clip pulse oximetry and arterial blood gas values were analysed. Three minutes after sensor placement, there were no significant differences between ear probe (cutaneous) and arterial carbon dioxide tensions (p = 0.367). Comparison of arterial with cutaneous carbon dioxide values demonstrated an excellent linear correlation (r2 = 0.92), and showed a standard error of estimate (SDEE) of 0.26 kPa (1.95 mmHg) only. The mean difference was 0.08 kPa (-0.60 mmHg) with a limits of agreement range of -0.38 kPa to +0.22 kPa (-2.85 mmHg to +1.65 mmHg). Concerning oxygen saturation measurements, the absolute SpO2 value deviated 1% or less from standard pulse oximetry. CONCLUSIONS: During general anaesthesia, postoperative recovery and critical care treatment, both monitoring of oxygenation and ventilation is important. Since pulse oximetry estimates only arterial oxygen saturation, periodic blood sampling is still necessary to determine the patient's arterial carbon dioxide status. We could demonstrate that the difference between cutaneous and arterial PCO2 was clinically unimportant, and therefore we conclude that the two methods of estimating the patient's carbon dioxide status may be used interchangeably. Our results demonstrated that 3 min after sensor placement, the new SpO2-PCO2 sensor prototype proved to be a reliable tool for continuous non-invasive monitoring of oxygenation and ventilation. PMID- 12578059 TI - An expert system for monitor alarm integration. AB - OBJECTIVE: Intensive care and operating room monitors generate data that are not fully utilized. False alarms are so frequent that attending personnel tends to disconnect them. We developed an expert system that could select and validate alarms by integration of seven vital signs monitored on-line from cardiac surgical patients. METHODS: The system uses fuzzy logic and is able to work under incomplete or noisy information conditions. Patient status is inferred every 2 seconds from the analysis and integration of the variables and a unified alarm message is displayed on the screen. The proposed structure was implemented on a personal computer for simultaneous automatic surveillance of up to 9 patients. The system was compared with standard monitors (SpaceLabs PC2), using their default alarm settings. Twenty patients undergoing cardiac surgery were studied, while we ran our system and the standard monitor simultaneously. The number of alarms triggered by each system and their accuracy and relevance were compared. Two expert observers (one physician, one engineer) ascertained each alarm reported by each system as true or false. RESULTS: Seventy-five percent of the alarms reported by the standard monitors were false, while less than 1% of those reported by the expert system were false. Sensitivity of the standard monitors was 79% and sensitivity of the expert system was 92%. Positive predictive value was 31% for the standard monitors and 97% for the expert system. CONCLUSIONS: Integration of information from several sources improved the reliability of alarms and markedly decreased the frequency of false alarms. Fuzzy logic may become a powerful tool for integration of physiological data. PMID- 12578060 TI - A graphical object display improves anesthesiologists' performance on a simulated diagnostic task. AB - OBJECTIVE: This study tests the hypothesis that a graphical object display (a data display consisting of meaningful shapes) will affect the ability of anesthesiologists to perform a diagnostic task rapidly and correctly. The diagnostic tasks studied were recognition and differentiation of five etiologies of shock--anaphylaxis, bradycardia, myocardial ischemia, hypovolemia, pulmonary embolus. METHODS: Data sets consisting of HR, Systemic Arterial BP, Pulmonary Arterial BP, CVP, and Cardiac Output were generated for five shock states and five non-shock states. The resulting 10 data sets were presented on a computer monitor to study subjects twice (first in an alpha-numeric format and then in the object format) for a total of twenty decision screens. Subjects used soft-buttons on a computer touch-screen monitor to: a) advance to the next display; b) differentiate a non-shock state from a shock state; and, c) select the etiology of shock state represented by the display (Figure 2). Data collection was automatic, using the internal clock and memory of the computer. RESULTS: Eleven anesthesiologists participated in this study. They completed a total of 3060 diagnostic decisions, half with each display format. Performance measures were time to decision and diagnostic accuracy. The object display improved no-shock recognition by 1.0 second and shock etiology determination by 1.4 seconds (p < 0.05). The object display also significantly improved accuracy for shock recognition by 1.4% and etiology determination by 4.1% (p < 0.05). Testing was completed in a time interval of <45 min per 10 trials. CONCLUSIONS: The primary finding of this study was that anesthesiologists using the object display format committed significantly fewer diagnostic errors when interpreting physiologic data. In addition, both the recognition of no-shock and the diagnosis of shock etiology were completed more rapidly when the object display was used. The major limitation of this initial trial is the simplicity of the test. Future investigation of the impact of the display on clinical decision making will require more realistic clinical scenarios with partial or full simulation to better understand the potential clinical impact. PMID- 12578061 TI - Comparison of statistical methods to predict the time to complete a series of surgical cases. AB - We present a statistical model for predicting the time to complete a series of successive, elective surgical cases. The use of sample means of case times and turnover times when scheduling cases does not minimize the operating room labor costs associated with errors in predicting times to complete series of cases. The problem of minimizing associated labor costs (both under and over utilization) can be converted to the problem of least absolute deviation regression. The dependent variables are the times to complete series of cases. The independent variables are the numbers of cases in each series that are in various categories (i.e., combinations of scheduled procedures and surgeons). Although the computational method is preferred on theoretical grounds to that involving sample means, application of both methods shows that the more practical method is to use the sample means of previous case times and turnovers. PMID- 12578062 TI - Intraoperative diagnosis of aortic pseudoaneurysm with transesophageal echocardiography. AB - A pseudoaneurysm of the ascending aorta is a rare complication of aortic valve endocarditis that requires prompt diagnosis. Several imaging strategies can be used; however, transesophageal echocardiography (TEE) has been utilized more frequently due to its superior resolution in detection of aortic valve complications. This case presents a patient with prosthetic valve dysfunction in which intraoperative TEE was used to diagnose a previously undetected aortic pseudoaneurysm, thus leading to a change in surgical management. PMID- 12578063 TI - Observations on use of wrong agent in an anesthesia agent vaporizer. AB - The creation of agent mixtures from the addition of the wrong agent to a vaporizer might pose a risk to the patient. Patient injury would be more likely if the anesthesia gas monitor displayed erroneous concentration values. Conventional inhalation agent monitors do not necessarily distinguish anesthetic agents. Some modern monitors have that ability but its clinical significance has not been determined. We wanted to simulate such an erroneous mixture in a laboratory setup. Six comparisons were made. Isoflurane, Enflurane, and Halothane vaporizers were first filled with the correct agent. They were run at 5 liters/minute fresh oxygen flow at a vaporizer dial setting of 5% until it reached the "refill" line. Then, one of two incorrect agents was added to the "full" line. Thereafter, the vaporizer continued at the same flow and the same dial setting until it was exhausted. Vaporizer output was recorded or calculated by using three methods of measurement: mass spectrometry, conventional infrared analysis (at 3.3 micrometer wave length), and piezoelectric crystal analysis. Additional calculations were used to estimate measurements that could not be made because of lack of available equipment. In a Halothane vaporizer: Enflurane added -not a significant problem; Isoflurane added--not a significant problem. In an Isoflurane vaporizer: Halothane added--not a significant problem; Enflurane added -not a significant problem. In an Enflurane vaporizer: Isoflurane added--not a significant problem; Halothane added--the sum of the delivered Halothane MAC and the delivered Enflurane MAC was twice the expected Enflurane MAC output from vaporizer, with conventional agent monitor reading which showed decreasing agent concentration. Patient injury could be more likely in this last case. In this last case and in all cases, piezoelectric crystal monitoring correctly displayed the sum of the two agent concentrations in volumes percent. Automatic agent identification can identify erroneous agents. PMID- 12578064 TI - Clinical competence assessment. PMID- 12578065 TI - Computerisation and decision making in neonatal intensive care: a cognitive engineering investigation. AB - This paper reports results from a cognitive engineering study that looked at the role of computerised monitoring in neonatal intensive care. A range of methodologies was used: interviews with neonatal staff, ward observations, and experimental techniques. The purpose was to investigate the sources of information used by clinicians when making decisions in the neonatal ICU. It was found that, although it was welcomed by staff, computerised monitoring played a secondary role in the clinicians' decision making (especially for junior and nursing staff) and that staff used the computer less often than indicated by self reports. Factors that seemed to affect staff use of the computer were the lack (or shortage) of training on the system, the specific clinical conditions involved, and the availability of alternative sources of information. These findings have relevant repercussions for the design of computerised decision support in intensive care and suggest ways in which computerised monitoring can be enhanced, namely: by systematic staff training, by making available online certain types of clinical information, by adapting the user interface, and by developing intelligent algorithms. PMID- 12578066 TI - Representation and classification of breath sounds recorded in an intensive care setting using neural networks. AB - OBJECTIVE: Develop and test methods for representing and classifying breath sounds in an intensive care setting. METHODS: Breath sounds were recorded over the bronchial regions of the chest. The breath sounds were represented by their averaged power spectral density, summed into feature vectors across the frequency spectrum from 0 to 800 Hertz. The sounds were segmented by individual breath and each breath was divided into inspiratory and expiratory segments. Sounds were classified as normal or abnormal. Different back-propagation neural network configurations were evaluated. The number of input features, hidden units, and hidden layers were varied. RESULTS: 2127 individual breath sounds from the ICU patients and 321 breaths from training tapes were obtained. Best overall classification rate for the ICU breath sounds was 73% with 62% sensitivity and 85% specificity. Best overall classification rate for the training tapes was 91% with 87% sensitivity and 95% specificity. CONCLUSIONS: Long term monitoring of lung sounds is not feasible unless several barriers can be overcome. Several choices in signal representation and neural network design greatly improved the classification rates of breath sounds. The analysis of transmitted sounds from the trachea to the lung is suggested as an area for future study. PMID- 12578067 TI - Computer assisted physiologic monitoring and stability assessment in vascular surgical patients undergoing general anesthesia--preliminary data. AB - BACKGROUND: Physiologic monitors present an influx of numerical data that can be overwhelming to the clinician. We combined several parameters in an effort to reduce the amount of information that must be continuously monitored including oxyhemoglobin saturation by pulse oximetry, end-tidal CO2 concentration, arterial blood pressure, and heart rate into an integrated measure--the health stability magnitude (HSM). The HSM is computed for a predetermined basal period, the reference HSM (RHSM), and recalculated continuously for comparison with the baseline value. In this study we present the HSM concept and examine changes in the HSM during abdominal aortic aneurysm surgery. MATERIALS AND METHODS: After IRB approval, nine patients were studied. The anesthesiologist recorded all significant intra-operative events. Within a defined time interval, data were recorded and used to calculate a combined parameter, the HSM. The baseline or reference value of this index (RHSM) was calculated after the induction of anesthesia. Individual HSM values were repeatedly calculated for ten second periods after the RHSM value was established. A > 30% deviation of the HSM from the RHSM was considered significant. Deviations in the HSM were compared with events recorded by the anesthesiologist on a paper record and with the record from an electronic record-keeping system. The deviation observed between two consecutive HSMs, called dHSM, was plotted against HSM to construct a contour diagram of data from all patients to which individual cases could be compared. RESULTS: The plot showed that dHSM vs. HSM values were tightly clustered. The inner contour on the distribution plot contained 90% of values. Individual patient's time course, projected on this diagram, revealed deviations form "normal" physiology. Fifty-nine events led to > 30% deviations in the HSM; 27 were anticipated events and 32 were unanticipated. CONCLUSION: The correlation between HSM and dHSM depicts changes in multiple monitored parameters that can be viewed using a single graphical representation. Projection of individual cases on the contour diagram may help the clinician to distinguish relative intraoperative stability from important events. Data reduction in this manner may guide clinical decision-making in response to unanticipated or unrecognized events. PMID- 12578068 TI - Systolic blood pressure at end-expiration measured by the automated systolic pressure variation monitor is equivalent to systolic blood pressure during apnea. AB - OBJECTIVE: It is necessary to define a reference systolic arterial blood pressure (RP) to calculate delta-Up (dUp) and delta-Down (dDown) for systolic pressure variation. Most studies define the reference pressure as the average systolic blood pressure during a short period of apnea. We describe an automated systolic pressure variation monitor that measures airway pressure and defines the reference pressure as the systolic blood pressure at end-expiration. The present study compares the reference systolic blood pressure measured at end-expiration by the automated systolic pressure variation monitor and the reference systolic blood pressure measured during apnea to test whether the end-expiration value is an adequate substitute for the value during apnea. METHODS: After obtaining informed consent, 108 sets of measurements of systolic pressure variation (SPV) were made in 20 intubated, mechanically-ventilated, anesthetized patients by the automated SPV monitor and during apnea. Measurements were taken during periods of hemodynamic stability defined as three consecutive end-expiratory systolic blood pressures within four mmHg of each other. The three systolic pressures at end expiration were averaged (RPmonitor). Immediately following these measurements, the ventilator was turned off and the systolic blood pressure was measured at 6, 8, 10 and 12 seconds of apnea. The reference pressure during apnea (RPapnea) was defined as the average of the systolic blood pressure at 8, 10 and 12 seconds of apnea. For each measurement set, RPmonitor and the systolic blood pressure at 6 seconds of apnea (SBP6) were compared to RPapnea using Bland-Altman analysis. RESULTS: Bland-Altman analysis for the difference between SBP6 and RPapnea yielded a small bias of -0.3 mmHg with standard deviation of 1.3, indicating that the systolic pressure tends to continue to increase slightly after 6 seconds of apnea. Results were similar for the difference between RPmonitor and RPapnea ( 0.2 +/- 3.1 mmHg). CONCLUSIONS: dUp and dDown are calculated using the reference pressure. RPmonitor is an average of 0.2 mm Hg less than RPapnea, thus dUp calculated by the automated SPV monitor is an avenge of 0.2 mm Hg greater than dUp measured by the reference pressure during apnea and dDown is 0.2 mm Hg less. Since the bias of -0.2 mmHg is clinically insignificant, there is acceptable agreement between the reference pressure obtained during apnea and that obtained by the automated SPV monitor at end-expiration. The mean difference between RPmonitor and RPapnea is explained by the continued rise in systolic pressure during the period of apnea as demonstrated by the difference between SBP6 and RPapnea. PMID- 12578069 TI - A proposed method for the measurement of anesthetist care variability. AB - OBJECTIVE: Some critical events in anesthesiology occur as seemingly preventable misadventures, their exact origins indeterminable. Inexperienced anesthetists, anesthesia machine malfunctions, lack of vigilance and human error inevitably initiate some incidents. Anesthesia training improves recognition and decision making. Avoiding crisis initiation and amelioration of those that do occur is one role of the consultant anesthesiologist. Safe patient care requires medical and procedural knowledge, technical expertise, and control of resources in a complex milieu. Anesthesia simulators are clinical laboratories where anesthetists can sharpen both cognitive and manual skills. Dynamic scenarios allow opportunities for anesthetists to explore and experience crises as they develop and apply their knowledge while attempting to manage these events. Simulator-based scenarios are reproducible and large amounts of useful data can be collected and saved. The authors hypothesize these data can be utilized to compare performance of anesthetists and to measure improvement of individual anesthetists over time. METHODS: We have designed "Stable Anesthesia," a prototypic scenario to test anesthetists' capabilities under the stress of performance guidelines. Three subjects performed anesthesia using the simulator and this protocol. Data from the simulator were archived by the system and analyzed by the authors. RESULTS: A simple mathematical analysis gave good separation of data from three subjects of different training level. CONCLUSIONS: It is suggested that the use of the techniques mentioned here may be of value in the development of a standardized testing protocol for anesthetists. PMID- 12578070 TI - Non-invasive cardiac output monitoring by aortic blood flow measurement with the Dynemo 3000. AB - The operating principles and methods for the continuous determination of aortic blood flow (ABF) with the Dynemo 3000 system are described in detail. The system uses a novel transesophageal ultrasonic Echo-Doppler probe simultaneously to measure aortic diameter and blood flow velocity at the same anatomic level, in real-time. Non-invasive ABF measurement is combined with vital sign data from standard monitors to provide a composite hemodynamic profile including volume, afterload and contractility data used by the physician to optimize therapy. A review of the clinical validation and comparison to thermodilution measurements showing a significant positive correlation over a wide range of clinical flow situations is also briefly presented. PMID- 12578071 TI - Evaluation of two new ecological interface approaches for the anesthesia workplace. AB - OBJECTIVE: Currently, vital parameters are commonly displayed as trends along a timeline. However, clinical decisions are more often based upon concepts, such as the depth of anesthesia, that are derived by combining parameter relationships and additional context information. The current displays do not visualize such concepts and therefore do not optimally support the decision process. A new display should present an ecological interface (EI). The principle of EI design is to visualize all of the information necessary for decision making in one single display. METHODS: In the first approach, we developed an EI that visualizes 35 relevant parameters for anesthesia monitoring. All of the parameters are generated by an anesthesia software simulator. Sixteen anesthetists had to administer two simulated general anesthetics: in one setting working only with the simulator's monitors ("Sim Only"), and in another setting working with the simulator's monitors in combination with the EI ("Combi1"). During each experiment, one unexpected critical incident (either blood loss or a cuff leakage) had to be identified. The control and monitoring behavior was analyzed by recording the subjects' eye movements and think-aloud protocol. With the help of the eye-tracking results, we re-designed the EI. The new EI was then tested with no eye tracking ("Combi2") on eight anesthetists under analogous conditions as in "Combi1." RESULTS: Cuff leakage was identified significantly quicker in "Combi1" (7 of 8 cases; time (T): 65 s +/- 73 s) than in "SimOnly" (6 of 8 cases; T: 222 s +/- 187 s). Blood loss was identified in 5 of 8 cases (T: 215 s +/- 76 s) in "Combi1" as quickly as in "SimOnly" (all cases; T: 217 s +/- 72 s). In "Combi1," the EI was used as the main source of information (in 43 +/- 19% of time) and was frequently favored when identifying an evolving critical incident. In "Combi2," cuff leakage was identified in 7 of 8 cases (T: 70 s +/- 111 s) as quickly as in "Combi1." Blood loss was identified significantly quicker in all cases (T: 147 s +/- 62 s) in "Combi2" than in "Combi1" and in "SimOnly." CONCLUSION: The results have shown that appropriately designed EIs may improve the anesthetist's decision making and focus attention on specific problems. Now, the findings have to be tested in future studies by widening the scope using other simulated scenarios and being closer to reality under real conditions in the OR. Eye tracking proved to be a useful method to analyze the anesthetists' decision making and appropriately re-design interfaces. PMID- 12578072 TI - Accuracy of capnography with a 30 foot nasal cannula for monitoring respiratory rate and end-tidal CO2 in children. AB - We tested the accuracy of a low flow (50 cc/min) sidestream capnography system equipped with an experimental 30-foot nasal cannula to monitor ventilatory status in children. End-tidal CO2 and respiratory rate, both at room air and in the presence of supplemental oxygen, were recorded simultaneously from the experimental 30-foot nasal cannula and the standard, FDA approved, 10-foot nasal cannula. The 30-foot nasal cannula was as accurate as the 10-foot nasal cannula in measuring respiratory rate and end-tidal CO2 in children. When supplemental oxygen was delivered by face-mask, there was no dilutional effect on the respiratory rate or end-tidal CO2 recorded with either the 10-foot or 30-foot nasal cannulas in place. PMID- 12578073 TI - Non-invasive estimation of cardiac output in critical care patients. AB - OBJECTIVE: This study was carried out to compare cardiac output measurements determined by thermodilution and by Portapres, a non-invasive system. DESIGN, PATIENTS AND SETTING: Eighty-seven non-invasive blood pressure measurements were performed in 46 patients in our critical care unit utilising the new, non invasive Portapres system. Cardiac output values were obtained from these blood pressure values using an aortic impedance model and compared to cardiac output values estimated by the thermodilution technique. MEASUREMENTS AND MAIN RESULTS: Statistically significant (p < 0.01) differences (2.3 l/min; limits of agreement +/-5 l/min) were noted between invasive and non-invasive cardiac output measurements. Differences in measured cardiac outputs increased for patients receiving catecholamine therapy, in patients with hemodynamic instability (e.g., sepsis and cardiac insufficiency), in patients with artificial ventilation, in patients with long duration of intensive care, in younger (<60 yr) patients and in women. We found no influence of the body mass index (BMI) on the accuracy of Portapres results. In only one single subgroup, 10 patients with pulmonary diseases, Portapres measurements were not statistically significant different from reference results. CONCLUSIONS: To date, Portapres measurements cannot replace thermodilution cardiac output estimations. Fluctuations of finger arterial perfusion due to hemodynamic instability, hypothermia and catecholamines may be responsible for problems of Portapres use in critically ill patients. PMID- 12578074 TI - Warning devices for prevention of dental injury during laryngoscopy. Preliminary report. AB - The prototypes of two novel warning devices for protection against dental trauma during direct laryngoscopy are described. Either can be attached to a standard laryngoscope blade (e.g., Macintosh). The basic design is that of a switch that is closed by pressure on the maxillary incisors by the laryngoscope blade. PMID- 12578075 TI - Teaching sedation and analgesia with simulation. AB - OBJECTIVE: This study reports on the efficacy of using the anesthesia simulator to teach sedation and analgesia to nurses. This provision of sedation and analgesia to a patient is accomplished with the goal of maintaining the ability of the patient to respond purposefully to auditory or tactile stimuli. METHODS: Nurses working in areas of the hospital where conscious sedation is performed were the participants in this sedation and analgesia training course. Prior to the training session, the participants read the American Society of Anesthesiology Practice guidelines for sedation and analgesia by non anesthesiologists. At the time of the training session, each participant completed a written pretest, had an introduction to sedation and analgesia with four clinical crisis teaching scenarios using the anesthesia simulator, a practical exam using the simulator, and a written post-test. Each participant was also given the opportunity to complete an evaluation of the session. RESULTS: Twenty nurses completed the training session. The written tests had a maximum possible score of 30. Mean score on the written pretest was 22.9 +/- 3.54, and mean score on the written post-test was 26.0 +/- 4.24 (p < 0.001). Seventeen of the twenty subjects scored higher on the post-test. Mean practical exam score was 5.5 of a possible 6.0. Mean participant rating of the education session was 3.75 (1 = poor, 4 = excellent). All but one participant rated the length of the training session as "about right." CONCLUSIONS: The anesthesia simulator provides an excellent tool for teaching conscious sedation skills to hospital nurses. The participants' test performance improved following the session, and they also rated the educational experience as excellent. PMID- 12578076 TI - Effects of digital filtering on the parameters of impulse-like eye movements. AB - Influence of two filtering modes were researched with electro-oculographically recorded impulse-like eye movements. Linear finite impulse response (FIR) and non linear hybrid median filters were explored by considering gain and latency parameters that yield the most important information in the case of these eye movements. It is stated that carefully selected lowpass filtering can securely be run without considerable changes in parameter values in order to discard noise stemming from physiological or other reasons. PMID- 12578077 TI - A case study in designing speech interaction with a patient monitor. AB - OBJECTIVE: New developments in speech interaction technology warrant the assumption that some of the interaction problems at anesthesia workplaces can be solved using speech interaction. One application might be the documentation of the anesthetic procedure. METHOD: To investigate this hypothesis, a speech input system was developed for a commercially available patient monitor (Datex-Ohmeda AS/3 Anesthesia Monitor with Record-Keeper), which in addition to standard functions also facilitates the generation of the anesthesia record. The speech input system was based on the IBM ViaVoice speech recognition technology (version Millennium Pro). The speech input device was preliminarily tested by 5 anesthetists wearing surgical masks under laboratory conditions and through retrospectively documenting 4 to 5 anesthetic procedures (each 12 to 17 documentation entries) in a calm situation. The speech interactive documentation procedure was compared with the conventional keyboard documentation procedure. RESULTS: The test persons achieved an overall word recognition rate between 72% and 92.4%. Between 76.5% and 97.7% of the single word commands could be completed without any corrections. The same qualifier held for 58.5% to 86.3% of the two- and three-word commands. Despite high speech recognition error rates, almost all documentation entries (236 of 238) could be finished using speech interaction. Comparing both documentation techniques, speech interaction was said to be more intuitive and caused less frustration than the keyboard documentation procedure. However, speech interactive documentation was rated less controllable because of the high speech recognition error rates and bccause the auditive feedback dialogues took too long. CONCLUSIONS: Modern speech recognition tools are still not advanced enough to facilitate the design of applications with an almost natural speech interface and widespread user acceptance. Nevertheless, many tasks in anesthesia have the necessary characteristics to be optimally supported by speech interaction. In contrast to earlier approaches to speech-interactive anesthesia workplaces, successful application today depends on the question of design rather than solely on that of technology. Many of the constraints and drawbacks of current technology can be overcome through appropriate design measures. The goals must focus first on identifying task areas in intensive care where speech-interaction can yield real benefit in terms of work efficiency, and second on developing and evaluating an ergonomic design of speech interaction. The intended users seem to look forward to the incorporation of speech interaction at the workplace. PMID- 12578078 TI - Monitoring of respiratory rate in postoperative care using a new photoplethysmographic technique. AB - OBJECTIVE: Photoplethysmography (PPG) is a non-invasive optical technique that measures variations in skin blood volume and perfusion. The PPG signal contains components that are synchronous with respiratory and cardiac rhythms. We undertook this study to evaluate PPG for monitoring patients' respiratory rate in the postoperative care unit, using a new prototype device. We compared it with the established technique, transthoracic impedance (TTI). METHODS: PPG signals from 16 patients (ASA classes 1-2, mean age 43 years) who were recovering from general anaesthesia after routine operations were recorded continuously for 60 minutes/patient. The respiratory synchronous part of the PPG signal was extracted by using a bandpass filter. Detection of breaths in the filtered PPG signals was done both visually and by using an automated algorithm. In both procedures, the detected breaths were compared with the breaths detected in the TTI reference. RESULTS: A total of 10,661 breaths were recorded, and the mean +/- SD respiratory rate was 12.3 +/- 3.5 breaths/minute. When compared with TTI, the rates of false positive and false negative breaths detected by PPG (visual procedure) were 4.6 +/- 4.5% and 5.8 +/- 6.5%, respectively. When using the algorithm for breath detection from PPG, the rates of false positive and false negative breaths were 11.1 +/- 9.7% and 3.7 +/- 3.8%, respectively, when compared to TTI. Lower respiratory rates increased the occurrence of false-positive breaths that were detected by the PPG using visual identification (p < 0.05). The same tendency was seen with the automated PPG procedure (p < 0.10). CONCLUSIONS: Our results indicate that PPG has the potential to be useful for monitoring respiratory rate in the postoperative period. PMID- 12578079 TI - Fire in the operating room. PMID- 12578080 TI - Optimization of alarms: a study on alarm limits, alarm sounds, and false alarms, intended to reduce annoyance. AB - Alarms in the operating room remain a major source of annoyance and confusion. Nearly all alarms result from a transgression of certain alarm limits. We surveyed manufacturers at a major meeting of anesthesiologists for their default alarm limits. We also conducted a mail survey of anesthesiologists in the United States, Finland, and The Netherlands, to learn their chosen alarm limits, reasons for turning off the alarms, and estimates of the number of false alarms. The surveys of medical monitoring manufacturers demonstrated a wide variety of default alarm settings. Anesthesiologists' chosen alarm limits tended to parallel the manufacturers' defaults. Fewer than 30% of anesthesiologists stated that they did not turn off the alarms. The leading reason for turning off alarms was the large number of false alarms. Estimates of the number of false alarms varied from 30% to 76%. The desired role of alarms in anesthetic practice and some suggestions for the general improvement of alarms are discussed. PMID- 12578081 TI - A meta-analysis of studies using bias and precision statistics to compare cardiac output measurement techniques. AB - INTRODUCTION: Bias and precision statistics have succeeded regression analysis when measurement techniques are compared. However, when applied to cardiac output measurements, inconsistencies occur in reporting the results of this form of analysis. METHODS: A MEDLINE search was performed, dating from 1986. Studies comparing techniques of cardiac output measurement using bias and precision statistics were surveyed. An error-gram was constructed from the percentage errors in the test and reference methods and was used to determine acceptable limits of agreement between methods. RESULTS: Twenty-five articles were found. Presentation of statistical data varied greatly. Four different statistical parameters were used to describe the agreement between measurements. The overall limits of agreement in studies evaluating bioimpedance (n = 23) was +/-37% (15 82%) and in those evaluating Doppler ultrasound (n = 11) +/-65% (25-225%). Objective criteria used to assess outcome were given in only 44% of the articles. These were (i) limits of agreement approaching +/-15-20%, (ii) limits of agreement of less than 1 L/min, and (iii) more than 75% of bias measurements within +/-20% of the mean. Graphically, we showed that limits of agreement of up to +/-30% were acceptable. CONCLUSIONS: When using bias and precision statistics, cardiac output, bias, limits of agreement, and percentage error should be presented. Using current reference methods, acceptance of a new technique should rely on limits of agreement of up to +/-30%. PMID- 12578082 TI - The effect of maintaining a constant preload or a constant degree of thumb abduction in the isometric twitch force of the thumb. AB - OBJECTIVE: To investigate the effects of maintaining a constant preload and of maintaining a constant degree of thumb abduction on the isometric twitch force during mechanomyography of the thumb, we monitored neuromuscular function in patients anaesthetized without the use of a neuromuscular blocking agent. In addition, we studied the relationship between the degree of thumb abduction, twitch force and preload. METHODS: Fourteen patients were divided randomly into two groups, determining the sequence of the experiments in respect of correcting the preload and maintaining a constant degree of abduction after allowing the twitch forces to stabilize. Both experiments lasted 15 minutes. In both groups the relationship between the degree of thumb abduction, twitch force and preload was studied. RESULTS: We found a progressive increase in twitch force both with (6.6%; CI: 3.7-9.3%; p < 0.001) and without (1.9%; CI: 0.4-3.5%; p = 0.020) continuous correction of the thumb preload. A significant greater increase in twitch force was seen when the preload had been corrected than when it was not (4.7%; CI: 0.8-8.7%; p = 0.023). In addition, both twitch force and preload appeared to depend on the degree of thumb abduction both before and at the time of measurement. CONCLUSIONS: Changes in length of the contracting muscle fibres and creep phenomena in the connective tissue of the muscles, both leading to changes in the sarcomere length of the muscle fibres, may explain the observations in this study. In general, a stabilized preload at a constant degree of abduction seems to be required in order to obtain a stable twitch force. PMID- 12578084 TI - A strategy for developing practice guidelines for the ICU using automated knowledge acquisition techniques. AB - OBJECTIVES: To implement practice guideline entry tools in a reminder system in order to provide decision support to health care workers in clinical care and emergency care environments. To design a knowledge acquisition environment that enables physicians to formulate, update, and verify guidelines without the assistance of a knowledge engineer. METHODS: We developed a knowledge acquisition environment for the Intensive Care Unit (ICU) consisting of 1) a graphical knowledge acquisition tool, 2) tools that perform logical and semantic tests on proposed guidelines, 3) a Patient Data Management System (PDMS) containing clinical patient data, and 4) an expert system that reminds ICU health care workers of inconsistencies between a treatment plan and implemented guidelines. Physicians enter the guidelines using the knowledge acquisition tool, after which consistency and correctness tests are performed on the guidelines. The guidelines are then transferred to the knowledge base of the reminder system and validated by applying the new guidelines to a large stored data set of previous patients. If the new guidelines are approved, they are exported to the reminder system that is used in daily practice. RESULTS: ICU physicians used the knowledge acquisition tool to enter 58 guidelines into the reminder system's knowledge base. These guidelines were tested on a data set consisting of 803 previously admitted patients. As a result, 27 guidelines fired at least once, generating 406 reminders in total. Of the 406 generated reminders, 356 (88%) were issued correctly and 50 (12%) were false alarms. The reminders that were issued correctly involved 3 situations: 1) the database contained inconsistent or incomplete information, 2) the actions or decisions of the health care workers were not the most appropriate ones, and 3) there was a potential risk involved. All false alarms were caused by the fact that the corresponding guidelines were not specific enough to handle certain exceptions. As a result of this analysis, the guidelines could be improved in such a way as to eliminate all false alarms. CONCLUSIONS: These first results demonstrate that this bottom-up knowledge acquisition strategy, implemented by the automated knowledge acquisition tools, enables medical specialists to improve the quality of computer support in an ICU without assistance of a knowledge engineer. PMID- 12578083 TI - The calibration and validation of a phase-modulated near-infrared cerebral oximeter. AB - OBJECTIVE: This study was undertaken to compare the cerebral oxygenation measured by an experimental phase-modulated near-infrared (NIR) spectroscopy system with capillary saturation estimated from jugular venous oxygen saturation. METHODS: Jugular venous catheters were placed in 30 patients undergoing carotid endarterectomy and 194 measurements of venous oxygen saturation were obtained intra operatively. Simultaneous measurement of optical path length at 754, 785, and 816 nm was performed using a phase-modulated near-infrared spectroscopy system. Optical calibration was performed using both an optical bench and a scattering mold. Hemoglobin saturation was calculated from NIR measurements using equations derived from diffusion theory. Capillary saturation was calculated from the arterial and venous saturations. RESULTS: Jugular venous saturations ranged from 41 to 92%. When calibrated using the optical bench, the NIR estimates of hemoglobin saturation deviated from estimated capillary values by an average of 2.6% bias and 4.3% deviation. No systematic bias was noted. NIR values derived from mold calibration were less accurate and precise (4.6% bias and 6.9% deviation.) Use of the initial venous sample as an in vivo calibration improved the accuracy of the mold calibration but did not alter the performance of the bench calibration. CONCLUSIONS: Under the conditions tested, an experimental phase-modulated near-infrared spectroscopy system calibrated using an optical bench agreed with capillary saturation estimated from jugular venous samples. Further work is necessary to demonstrate valid performance of the system under other conditions. PMID- 12578085 TI - Work of breathing-tidal volume relationship: analysis on an in vitro model and clinical implications. AB - OBJECTIVE: Work of breathing (WoB) is currently employed to assess the afterload on the respiratory muscles and to estimate the energy expenditure for breathing. Since WoB depends on the ventilated tidal volume (TV), WoB*L(-1), the indicized form of WoB has been employed as a measure of WoB which is independent of TV. Actually, the independence of WoB*L(-1) from the ventilated TV has never been demonstrated. The aim of this study was to verify the predicted TV-independence of WoB*L(-1) on an in vitro model. METHODS: Our experimental model was constituted as follows: two endotracheal tubes, with internal diameter measuring respectively 6.5 and 8.5 mm, were alternatively connected with two rubber balloons whose compliance was respectively 0.02 and 0.06 L/hPa; the system was mechanically ventilated at ten different tidal volumes, ranging from 0.3-1 l. Flow rate was kept constant (35 l/m) during the whole experiment. RESULTS: Both elastic components of the model showed a static volume-pressure relationship which was linear in the experimental range of TV. In all combinations of resistance and compliance WoB increased quadratically whereas WoB*L(-1) increased linearly with the growing TV (p < 0.001). CONCLUSIONS: These results demonstrate the TV-dependence of WoB*L(-1) and suggest that WoB*L(-1), if TV changes, cannot be considered as an index of respiratory muscle afterload and should not be used as a guide for weaning patients from the mechanical ventilation. Finally, we introduced a new parameter (WoB1L) which seems to be a more TV-independent measure of respiratory work. PMID- 12578086 TI - Evaluation of hemostasis by the Clot Signature Analyzer: a potentially valuable device for the anesthesiologist. AB - Rapid detection of hemostatic defects presents a challenge for the anesthesiologist who must balance anesthetic and surgical considerations for maintaining adequate platelet and coagulant factors, while keeping allogenic blood exposure to a minimum. The Clot Signature Analyzer, a point-of-care device capable of rapid response and easy interpretation is described here. Its applicability in two obstetrical patients with platelet dysfunction is discussed. PMID- 12578087 TI - Evaluation of a new advanced thoracic bioimpedance device for estimation of cardiac output. AB - OBJECTIVE: This study is an evaluation of a new thoracic bioimpedance cardiac output monitoring system which incorporates a modified form of the Kubicek equation and a method of estimating the left ventricular ejection time from the time derivative bioimpedance signals. METHODS: The performance of the new system was compared with conventional thermodilution in a porcine model. One hundred and ninety nine (n = 199) paired measurements of thermodilution cardiac output (TDCO) (range 1.20-18.00 L/min) and thoracic bioimpedance cardiac output (BICO) were collected in 7 pigs. The bioimpedance measurements were adjusted for the animal's weight and chest circumference, thus compensating for the differences in the anatomy of pigs when compared to humans. Data were compared using weighted correlation coefficient and Bland-Altman analysis. RESULTS: The weighted correlation coefficient between TDCO and BICO values was 0.87 (n = 199). The Bland-Altman technique yielded a precision of the device of +/-1.69 L/min with a bias of 0.11 L/min. CONCLUSION: The results from the porcine study show that the new system performed well over a wide range of cardiac outputs, comparing favorably with data from other new bioimpedance cardiac output devices currently available in the market. PMID- 12578088 TI - The first object oriented monitor for intravenous anesthesia. AB - OBJECTIVE: To describe the design and implementation of "INFUSION TOOLBOX," a software tool to control and monitor multiple intravenous drug infusions simultaneously using pharmacokinetic and pharmacodynamic principles. METHODS: INFUSION TOOLBOX has been designed to present a graphical interface. Object Oriented design was used and the software was implemented using Smalltalk, to run on a PC. Basic tools are available to manage patient, drugs, pumps and reports. These tools are the PatientPanel, the DrugPanel, the PumpPanel and the HistoryPanel. The screen is built dynamically. The panels may be collapsed or closed to avoid a crowded display. We also built control panels such as the Target ControlPanel which calculates the best infusion sequence to bring the drug concentration in the plasma compartment to a preset value. Before drug delivery, the user enters the patient's data, selects a drug, enters its dilution factor and chooses a pharmacokinetic model. The calculated plasma concentration is continually displayed and updated. The anesthetist may ask for the history of the delivery to obtain a graphic report or to add events to the logbook. A panel targeting the effect is used when a pharmacodynamic model is known. Data files for drugs, pumps and surgery are upgradable. DISCUSSION: By creating a resizeable ControlPanel we enable the anesthetist to display the information he wishes, when he wishes it. The available panels are diverse enough to meet the anesthetist needs; they may be adapted to the drug used, pumps used and surgery. It is the anesthetist who builds dynamically its different control screens. CONCLUSION: By adopting an evolutionary solution model we have achieved considerable success in building our drug delivery monitor. In addition we have gained valuable insight into the anesthesia information domain that will allow us to further enhance and expand the system. PMID- 12578089 TI - Effects of one-lung ventilation on cardiac autonomic nervous activity as evaluated by power spectral analysis of heart rate variability. AB - The purpose of this prospective study was to evaluate the effects of one-lung ventilation on the activity of the cardiac autonomic nervous system. Ten adult patients who underwent thoracotomy were endotracheally intubated with a double lumen tube under general anesthesia using isoflurane. After induction of anesthesia, a continuous, 256-sec electrocardiogram (ECG) was obtained during bilateral lung ventilation (control) followed by recordings during one-lung ventilation of each side. Using the R-R interval tachograms obtained for the 256 sec ECGs, low frequency (LF: 0.04-0.15 Hz) and high frequency (HF: 0.15-0.40 Hz) bands of the spectral density of the heart rate variability and the HF/LF ratio were analyzed using the fast Fourier transform algorithm. Log(HF), which indicates parasympathetic activity, increased during one-lung ventilation on each side, but did not differ between ventilated sides. Log(LF), which represents sympathetic and parasympathetic activity, increased similarly to log(HF) on both sides. Log(HF/LF), the balance of the sympathetic and parasympathetic activity, did not change during one-lung ventilation. We suggest that one-lung ventilation alone does not substantially affect the cardiac autonomic nervous system. PMID- 12578090 TI - Atypical "tails-up" capnograph due to breach in the sampling tube of side-stream capnometer. AB - OBJECTIVE: An atypical "tails-up" capnograph pattern was noticed in a patient during the use of an accidentally crushed sampling tube with a slit-like hole. We investigated the mechanics involved in the observed capnograph pattern. METHODS: Forty consenting ASA I patients of both sexes presenting for tonsillectomy were included in this study. After intravenous induction of anaesthesia, intermittent positive pressure ventilation (IPPV) using a mechanical ventilator was maintained for 20 min and the capnograph trace, ETCO2, and inspiratory/expiratory sevoflurane were compared using a breached sampling tube and then an intact sampling tube. Similar comparisons were made during spontaneous breathing. RESULTS: During IPPV, an atypical "tails-up" capnograph was noted using the breached sampling tubing. At similar inspiratory sevoflurane (2.0 +/- 0.03) levels, expiratory levels (0.9 +/- 0.03) were significantly lower when using the breached sampling tube than the intact tube (1.7 +/- 0.03). ETCO2 with the breached sampling tube (26.8 +/- 0.30 mmHg) showed significantly lower values than with the intact sampling tube (37 +/- 0.3). During spontaneous breathing, the capnograph was normal in shape with both sampling tubes, but ETCO2 and both insp./exp. sevoflurane levels were lower with the breached sampling tube. CONCLUSION: During IPPV, pressure in the breathing circuit is lower during exhalation, thus allowing air to enter through the slit-like hole in the sampling tube causing erroneously low ETCO2 and expiratory sevoflurane. With inspiration, positive pressure in the breathing circuit, transmitted to the sampling tube, prevents air admixture and the upsurge in CO2 is displayed giving the capnograph an atypical "tails-up" appearance. During spontaneous breathing, since pressure in the breathing circuit barely becomes positive during exhalation and is negative during inspiration, air mixes with the sampled gas during both phases and so the capnograph shape was normal but with lower values for ETCO2, insp./exp. sevoflurane, and nitrous oxide levels. If undiagnosed, this defect in the sampling tube can lead to significant errors in the measurement of inspired and expired gas concentrations. PMID- 12578091 TI - Measurement of endogenous nitric oxide production. AB - The measurement of exhaled pulmonary nitric oxide concentrations requires that contamination from the upper respiratory tract and inhaled gases be eliminated. This can be achieved with no risk in the clinical setting of intubated patients of all ages in the operating room or intensive care unit. Further modifications of the anesthetic/ventilatory circuit allow for accurate determination of tidal volume and minute ventilation. PMID- 12578092 TI - Local cardiac wall stabilization influences the reproducibility of regional wall motion during off-pump coronary artery pass surgery. AB - OBJECTIVE: Myocardial ischemia is a risk factor during off-pump coronary artery bypass procedures. The development of new regional wall motion abnormalities assessed by transesophageal echocardiography (TEE) is a very sensitive sign of myocardial ischemia. To facilitate anastomosis, the epicardial area of the anastomosis site is often immobilized by a "stabilizer." This study was designed to investigate whether cardiac wall stabilization with an epicardial stabilizer could affect the interpretation of wall motion during coronary anastomosis without cardiopulmonary bypass. METHODS: The TEE videotapes of 15 adult patients were investigated. Left ventricular (LV) transgastric short and long axis views were divided according to a modified 16-segment method. LV wall motion was scored using a 5-grade scale by two independent blinded investigators during pre occlusion, occlusion, and reperfusion of anastomosed coronary arteries. The wall motion scores of a stabilized segment combined with two adjacent segments were compared with those of non-stabilized segments. Interobserver agreement was assessed using the weighted kappa statistic. RESULTS: A total of 216 segments were analyzed by two investigators. The interobserver kappa coefficient in pre occlusion and reperfusion periods was 0.87, 0.87 and 0.86, 0.87, respectively, indicating high agreements without stabilizer. During the occlusion period in stabilized and non-stabilized segments, it was 0.59 and 0.76, respectively, showing significantly less reproducibility in the presence of stabilizer. CONCLUSION: Cardiac wall stabilization affects the reproducibility in the interpretation of regional wall motion during off-pump coronary artery bypass surgery. Caution should be used when monitoring for myocardial ischemia using TEE during coronary artery bypass surgery with epicardial stabilizer. PMID- 12578093 TI - An IBM PC-based system for the assessment of cardio-respiratory function using oscillating inert gas forcing signals. AB - OBJECTIVE: An IBM PC-based real-time data acquisition, monitoring and analysis system was developed for the assessment of cardio-respiratory function, i.e. airway dead space, alveolar volume and pulmonary blood flow, using oscillating inert inspired gas forcing signals. METHODS: The forcing gas mixture was generated by an in-house sinusoid gas delivery unit. The system interfaced with a mass spectrometer and an airway flow transducer, and performed real-time tracking of the breath-by-breath end-inspired, end-expired and mixed-expired concentrations. It calculated the cardiorespiratory parameters using two, i.e. continuous and tidal, in-house mathematical models of the lungs. The system's performance was evaluated using a mechanical bench lung, laboratory subjects and awake adults breathing spontaneously. Its predictive accuracy was compared with the measured volumes of the bench lung; single breath CO2 test for airway dead space and N2 washout for alveolar volume in laboratory subjects and awake adults; and thermal dilution technique for pulmonary blood flow in laboratory subjects. RESULTS: Close agreements were found between the true and predicted airway dead space, i.e. mean differences of -12.39%, 14.47% and -17.49%, respectively, and that of alveolar volume, i.e. -8.03%, -3.62% and 7.22%, respectively, in the bench lung, laboratory subject and awake adult studies; and that of pulmonary blood flow (-23.81%) in the laboratory subjects using the continuous lung model. Even closer agreements were observed for airway dead space (-5.8%) and alveolar volume (-4.01%) of the bench lung and for pulmonary blood flow (-8.47%) in the laboratory subjects using the tidal lung model. CONCLUSIONS: A system was developed to deliver, monitor and analyse on-line, and in real-time, output data from the sinusoid forcing technique. The technique was administered using the system in various subjects, and produced favourable predictions. PMID- 12578094 TI - Foundations of anesthesiology. An account of some hydraulic and hydrostatical experiments made on the blood and blood-vessels of animals. 1710 [classical article]. PMID- 12578095 TI - In my opinion: Regional anesthesia preferable for carotid surgery. PMID- 12578096 TI - Detection and treatment of cerebral hypoxia key to avoiding intraoperative brain injuries. PMID- 12578097 TI - Latex allergy: a problem for patients and personnel. PMID- 12578098 TI - Mega-dose lidocaine dangers seen in "tumescent" liposuction. PMID- 12578100 TI - Infrared transmission of electronic information via LAN in the operating room. AB - Recent advances in technology have brought many kinds of monitoring devices into the operating room (OR). The information gathered by monitors can be channeled to the operating ward information system via a local area network (LAN). Connecting patients to monitors and monitors to the LAN, however, requires a large number of cables. This wiring is generally inconvenient and particularly troublesome if the layout of the OR is rearranged. From this point of view, wireless transmission seems ideally suited to clinical settings. Currently, two modes of wireless connectivity are available: radio-frequency (RF) waves or infrared (IR) waves. Some reports suggest that RF transmission is likely to cause electromagnetic interference (EMI) in medical devices such as cardiac pacemakers or infusion pumps. The risk of malfunctioning life-sustaining devices and the catastrophic consequences this would have on seriously ill patients rules out the use of RF. Here, we report an IR system using IR modems for LAN connectivity in the OR. In this study, we focused on the possible detrimental effects of EMI during wireless connectivity. In our trial, we found no evidence of EMI of IR modems with any of the medical devices we tested. Furthermore, IR modems showed similar performance to a wired system even in an electrically noisy environment. We conclude that IR wireless connectivity can be safely and effectively used in ORs. PMID- 12578099 TI - A prototype device for standardized calibration of pulse oximeters. AB - OBJECTIVE: To develop and test a method for standardized calibration of pulse oximeters. METHODS: A novel pulse oximeter calibration technique capable of simulating the behavior of real patients is discussed. It is based on an artificial finger with a variable spectral-resolved light attenuator in conjunction with an extensive clinical database of time-resolved optical transmission spectra of patients fingers in the wavelength range 600-1000 nm. The arterial oxygen saturation of the patients at the time of recording was derived by analyzing a corresponding blood sample with a CO-oximeter. These spectra are used to compute the modulation of the light attenuator which is attached to the artificial finger. This calibration method was tested by arbitrarily playing back recorded spectra to pulse oximeters and comparing their display to the value they displayed when the spectra were recorded. RESULTS: We were able to demonstrate that the calibrator could generate physiological signals which are accepted by a pulse oximeter. We also present some experience of playing back recorded patient spectra. The mean difference between the original reading of the pulse oximeters and the display when attached to the calibrator is 1.2 saturation points (displayed oxygen saturation SpO2) with a standard deviation of 1.9 saturation points. CONCLUSIONS: The tests have shown the capabilities of a spectral light modulator for use as a possible calibration standard for pulse oximeters. If some improvements of the current prototype can be achieved we conclude from the experience with the device that this novel concept for the calibration of pulse oximeters is feasible and that it could become an important tool for assessing the performance of pulse oximeters. PMID- 12578101 TI - Elimination of cardiogenic oscillations in the capnograph by applying low positive end-expiratory pressure (PEEP). PMID- 12578103 TI - Estimation of jugular venous O2 saturation from cerebral oximetry or arterial O2 saturation during isocapnic hypoxia. AB - OBJECTIVE: Near-infrared spectroscopy (NIRS) has the potential for providing valuable information about oxygen delivery to the brain. However, questions have been raised about the accuracy of these measurements. This study was undertaken to compare noninvasive cerebral saturation measurements to jugular venous saturation under conditions of hypoxia and hypercapnia. METHODS: Data was obtain on forty-two subjects. Cerebral oxygenation was measured with a Somanetics INVOS 4100-SSA placed on the forehead of the subjects. PETCO2 was controlled to approximately 2 and 7 mmHg above resting values and PETO2 was controlled to 80, 45, 60 and 41 mmHg consecutively for four of five minutes each. Internal jugular blood gas measurements were made via a retrograde catheter. RESULTS: Both the cerebral oximetry measured saturation (rSO2) and the jugular venous saturation (SjvO2) were significantly increased by increasing the PETCO2 at all levels of hypoxia. The increase in the rSO2 was less than the increase in SjvO2. The rSO2 had a bias of 5.2% and a precision of 10.7% compared to the measured SjvO2. DISCUSSION: Cerebral oxygen saturation measured by cerebral oximetry compares well to the measured SjvO2 in normal subjects, despite multiple physiological reasons for differences. The closer relationship of SjvO2 to rSO2 than SaO2 under the conditions of these experiments indicates that the measurement reflects primarily intracranial saturation. However, outcome studies under clinical conditions are needed to determine the clinical utility of cerebral oximetry. PMID- 12578102 TI - Use of an anesthesia information management system (AIMS) to evaluate the physiologic effects of hypnotic agents used to induce anesthesia. AB - OBJECTIVE: The aim of this study was to utilize an anesthesia information management system (AIMS) in investigating the effects of hypnotic agents used to induce anesthesia on blood pressure, heart rate and arterial oxygen saturation. The characteristics of these agents, known from previous studies, were compared to the effects documented in this study during routine clinical use. METHODS: During the years 1997 and 1998, all relevant data from anesthetic procedures were recorded online using the automated anesthesia information system NarkoData. The data from 8,078 general anesthesia procedures using endotracheal intubation were exported via "structured query language" (SQL) from the AIMS database into a statistics program after excluding children (age < 14), patients who received atropine during induction and procedures with use of extracorporeal circulation. The effects of drug administration on systolic, diastolic and mean arterial blood pressure (SBP, DBP, MBP), heart rate (HR) and arterial oxygen saturation (SpO2) were analyzed prior to induction and at 5, 10 and 15 minutes following bolus administration of the hypnotic agent. The data were classified into three groups based on the induction agent used: thiopental, etomidate or propofol and further separated into two groups based on ASA status (ASA < or = II and ASA > II). The mean and standard deviations were calculated for each parameter at each point in time. Statistical comparisons were performed to determine whether the results for each time point differed from the previous time point. RESULTS: There was a significant decrease in blood pressure (MAP, SBP, DBP) after bolus administration of all three hypnotics in all of the 8,078 procedures analyzed. The decrease was greater in patients of ASA class > II than in those of ASA class < or = II. Propofol caused the greatest drop in blood pressure whereas etomidate caused the least. During the observation period the HR also fell in each group, except for thiopental where an initial rise of the HR could be observed. An initial rise of SpO2 was recorded in each group with no differences observed between the individual hypnotics. CONCLUSIONS: The effects of the induction hypnotic agents thiopental, etomidate and propofol on blood pressure and heart rate as documented by an AIMS corresponded to those found in clinical studies. An AIMS with the corresponding documentation, software and database structure is suitable for collecting and evaluating data for dinical investigations. PMID- 12578104 TI - Comparison of a prototype esophageal oximetry probe with two conventional digital pulse oximetry monitors in aortocoronary bypass patients. AB - OBJECTIVE: Pulse oximetry (SpO2) is the non-invasive standard for monitoring arterial oxygen saturation in patients undergoing anesthesia, but is subject to external interference by motion artifact, peripheral vasoconstriction, and low cardiac output. We hypothesized that oximetry signals could be acquired from the esophagus when peripheral pulse oximetry is unobtainable. Therefore, we tested an esophageal stethoscope which incorporates transverse oximetry photodetectors and emitters in patients undergoing coronary bypass surgery. METHODS: Immediately after induction of general anesthesia in 10 coronary artery bypass (CABG) patients, Criticare and Nellcor digital probes were positioned on the left hand, concurrent with placement of an esophageal SpO2 probe. A computer recorded 5,910 matched oximetry signals every 15 sec during an average of 2.5 hrs. All SpO2 measurements were before, and immediately after non-pulsatile, hypothermic cardiopulmonary bypass. Data represent the percentage (median value [range]) of the total monitored time that a SpO2 value was displayed. RESULTS: The Nellcor (99.8%, range 6.5-100%) and Criticare (99.7%, range 36.6-100%) acquired and displayed saturation signals more frequently (p = 0.003) than the esophageal monitor (75.3%, range 42.1-95.8%). The two standard digital oximeters had a mean difference of 0.9%, with a standard deviation of the differences of 0.9. The esophageal probe had a mean difference of -5.2% and -4.8%, with standard deviation of differences of 8.0 and 7.7 (compared to the Nellcor and Criticare monitors, respectively). A second-generation prototype shielded from electrocautery interference was tested in an additional 4 patients. The shielded prototype displayed signals more frequently (96.7%, range 68.4-100%) than the original esophageal prototype. CONCLUSIONS: Digital pulse oximetry failure is common in CABG patients, probably because of marginal cardiac output and peripheral vasoconstriction associated with hypothermia. Our study could not confirm that esophageal technology, which utilizes the esophagus as a site of transflectance oximetry, was superior to conventional digital pulse oximetry. PMID- 12578105 TI - Using an anesthesia information management system to prove a deficit in voluntary reporting of adverse events in a quality assurance program. AB - OBJECTIVE: A deficit is suspected in the manual documentation of adverse events in quality assurance programs in anesthesiology. In order to verify and quantify this, we retrospectively compared the incidence of manually recorded perioperative adverse events with automatically detected events. METHODS: In 1998, data of all anesthetic procedures, including the data set for quality assurance of the German Society of Anaesthesiology and Intensive Care Medicine (DGAI), was recorded online with the Anesthesia Information Management System (AIMS) NarkoData4 (Imeso GmbH). SQL (Structured Query Language) queries based on medical data were defined for the automatic detection of common adverse events. The definition of the SQL statements had to be in accordance with the definition of the DGAI for perioperative adverse events: A potentially harmful change of parameters led to therapeutic interventions by an anesthesiologist. RESULTS: During 16,019 surgical procedures, anesthesiologists recorded 911 (5.7%) adverse events manually, whereas 2966 (18.7%) events from the same database were detected automatically. With the exception of hypoxemia, the incidence of automatically detected events was considerably higher than that of manually recorded events. Fourteen and a half percent (435) of all automatically detected events were recorded manually. CONCLUSION: Using automatic detection, we were able to prove a considerable deficit in the documentation of adverse events according to the guidelines of the German quality assurance program in anesthesiology. Based on the data from manual recording, the results of the quality assurance of our department match those of other comparable German departments. Thus, we are of the opinion that manual incident reporting seriously underestimates the true occurrence rate of incidents. This brings into question the validity of quality assurance comparisons based on manually recorded data. PMID- 12578106 TI - The relationship between rate of administration of an intubating dose of rocuronium and time to 50% and 90% block at the adductor pollicis muscle. AB - OBJECTIVE: To determine the relationship between the rate of rocuronium injection and the onset time of neuromuscular block. METHODS: After intravenous induction, 60 female patients (ASA I-II) were assigned randomly into 3 groups for rocuronium administration within 1-15, 15-30 or 30-60 seconds. Acceleromyography of the thumb was performed using train-of-four (TOF) stimulation. Times to 50% and 90% twitch depression of the first twitch of the TOF stimulation (T1) were recorded. RESULTS: Injection time significantly influences time to 50% relaxation, but not time to 90% relaxation. Body mass index is negatively correlated with time to 50% and 90% relaxation. CONCLUSIONS: We conclude that rate of injection influences only the initial phase of development of the block and that slower injection times do not significantly affect time to 90% relaxation at the adductor pollicis muscle. PMID- 12578108 TI - Absent CO2 detection after tracheal intubation due to sampling line obstruction. PMID- 12578107 TI - Limits of corrected flow time to monitor hemodynamic status in children. AB - OBJECTIVE: Doppler corrected flow time (i.e., corrected left ventricular ejection time) as a noninvasive tool for assessing hemodynamic changes has been previously reported for adult patients. Its use in paediatrics seems to be worthwhile but no data concerning its accuracy are presently available in this population. The purpose of this work was to study the relationships between corrected flow time (FT) and indices of systemic vascular resistance (SVR) and of myocardial contractility in healthy children. METHODS: Twenty healthy children performed a graded maximal bicycle exercise in order to induce physiological hemodynamic alterations. Hemodynamic parameters were measured with an echocardiography Doppler at rest and within a few minutes of post exercise. Cycle time (RR), mean aortic flow velocity, mean systolic velocity (MSV), FT, peak velocity (PV), and stroke distance were measured on the Doppler aortic velocity waveform. Cardiac index (CI) and SVR were calculated from the classical volumetric equation. Corrected FT was calculated by using Bazett's formula (FTb = FT/square root(RR)) and a simplified formula FTc = FTmeasured + [1.29 x (HR - 60)]. RESULTS: Post exercise, SVR, RR, FT, decreased, while CI, PV and MSV increased and stroke distance remained unchanged. After multiple regression analysis no significant correlation between SVR and FTb and SVR or FTc was noted. A significant correlation appeared between FTb and, respectively, PV (r = -0.83; p < 0.001), stroke distance (r = 0.78; p < 0.001) and RR (r = -0.52; p = 0.0016). A significant correlation was also shown between FTc and, respectively, PV (r = 0.71; p < 0.001) and stroke distance (r = 0.63; p < 0.001) but not with RR. CONCLUSIONS: These results show that the use of Bazett's formula correct FT could lead to hemodynamic misinterpretations, because it does not rule out all the heart rate effect. Moreover, in healthy children corrected FT appears as an inaccurate index to monitor physiological afterload alterations, because of the involvment of other hemodynamic factors such as contractility in its variation. PMID- 12578109 TI - Landmark report published on patient safety. PMID- 12578110 TI - Let no patient be harmed by anesthesia. PMID- 12578111 TI - Danger from OR fires still a serious problem. ASA Panel reports risks. PMID- 12578112 TI - Orthopaedic surgeons issue advisory on wrong-site surgery. PMID- 12578113 TI - Particle characteristics responsible for effects on human lung epithelial cells. AB - Some recent epidemiologic investigations have shown an association between increased incidence of respiratory symptoms and exposure to low levels of particulate matter (PM*) less than 10 microm or less than 2.5 microm in aerodynamic diameter (PM10 and PM2.5, respectively). If particulates are causally involved with respiratory symptoms, it is important to understand which components may be responsible. However, increasing evidence suggests that transition metals present in particles, especially iron, generate reactive oxygen species (ROS) that may be involved in producing some of the observed respiratory symptoms. The hypothesis for this study is twofold: bioavailable transition metals from inhaled airborne particulates catalyze redox reactions in human lung epithelial cells, leading to oxidative stress and increased production of mediators of pulmonary inflammation: and the size, transition metal content, and mineral speciation of particulates affect their ability to cause these effects. This work focused on the relation between physical characteristics of particles (eg, size, bioavailable transition metal content, and mineral speciation) and their ability to generate hydroxyl radicals in cell-free systems and to cause oxidative stress, which results in the synthesis of mediators of pulmonary inflammation in cultured human lung epithelial cells. These relations were studied by comparing size-fractionated, chemically characterized coal fly ash (CFA) produced by combustion of three different coals to obtain milligram quantities of ash. One transition metal, iron, was studied specifically because it is by far the predominant transition metal in CFA. In addition, smaller quantities of particles from gasoline engines, diesel engines, and ambient air were studied. Phosphate buffer soluble fractions from particles from all sources were capable of generating ROS, as measured by production of malondialdehyde (MDA) from 2-deoxyribose. This activity was inhibited over 90% for all particles by the metal chelator N-[5-[3-[(5-aminopentyl)hydroxycarbamoyl]propionamidol pentyl]-3-[[5-(N-hydroxyacetamido)pentyl]carbamoyl]propionohydroxamic acid (desferrioxamine B, or DF), strongly suggesting that transition metal(s), probably iron, were responsible. Particles from coal or gasoline combustion had greater ability to produce ROS than particles from diesel combustion. Iron was mobilized by citrate (at pH 7.5) from particles of all sources tested; gasoline combustion particles were the only particles not analyzed for iron mobilization because there were not enough particles for the iron mobilization assay. CFA particles were size-fractioned; the amount of iron mobilized by citrate was inversely related to the size of particles and also depended on the source of coal. Iron from the CFA particles was responsible for inducing the iron-storage protein ferritin in cultured human lung epithelial cells (A549 cells). The amount of iron mobilized by citrate was directly proportional to the amount of ferritin induced in the A549 cells. Iron from the CFA was also responsible for inducing the inflammatory mediator interleukin (IL) 8 in A549 cells. Iron existed in several species in the fly ash, but the bioavailable iron was associated with the glassy aluminosilicate fraction, which caused ferritin and IL-8 to be induced in the A549 cells. In crustal dust, another component of urban particulates, iron was associated with oxides and clay but not with aluminosilicates. The crustal dust contained almost no iron that could be mobilized by citrate. Iron could be mobilized from diesel combustion particulates, but at a much lower level than for all other combustion particles. Samples of ambient PM2.5 collected in Salt Lake City over 5-day periods during one month varied widely in the amount of iron that could be mobilized. If bioavailable transition metals (eg, iron) are related to the specific biological responses outlined here, then the potential exists to develop in vitro assays to determine whether particulates of unknown composition and origin can cause effects similar to those observed in this study. PMID- 12578114 TI - An argument for Vitamin E supplementation in the management of systemic inflammatory response syndrome. AB - The systemic inflammatory response syndrome results from an uncontrolled, overexpression of the normal host inflammatory response, leading to destruction of host tissue and subsequent organ failure. Oxidant stress has been implicated in this process both as a mechanism for direct cellular injury, as well as activation of intracellular signaling cascades within inflammatory cells resulting in progression of the inflammatory response. Vitamin E is an inexpensive, nontoxic, chain-breaking antioxidant that has therapeutic potential in regulating this process. This review seeks to evaluate the current literature regarding the use of Vitamin E in controlling the excessive inflammation seen in systemic inflammatory response syndrome and argues for further study of its therapeutic potential for these critically ill patients. PMID- 12578115 TI - Promissory notes, demonstrations for the first time, and other claims in scientific research. PMID- 12578116 TI - Impairment of beta-adrenergic signaling in healthy peripheral blood mononuclear cells exposed to serum from patients with septic shock: involvement of the inhibitory pathway of adenylyl cyclase stimulation. AB - The objective of this study was to determine whether the serum of patients with sepsis could alter the capability of healthy human peripheral blood mononuclear cells (PBMC) to synthesize cAMP in response to beta-adrenergic stimulation and to evaluate the involvement of the inhibitory pathway (Gi) of adenylyl cyclase in the sepsis-induced alteration of beta-adrenergic signaling. First, PBMC from a healthy donor were incubated for 24 h in serum-containing medium according to three culture conditions: serum alone, serum with pertussis toxin, and serum with propranolol. Second, PBMC were stimulated with 10(-5) M isoproterenol or 10(-6) M forskolin, and measurement of cyclic adenosine monophosphate (cAMP) intracellular accumulation was performed. Serum samples were obtained from three groups of subjects: 14 patients with severe sepsis, 21 patients with septic shock, and 10 healthy control subjects. Basal and forskolin-stimulated cAMP levels were similar in PBMC cultured in control or in septic serum. Isoproterenol-stimulated accumulation was reduced in PBMC preincubated in septic serum. The lowest cAMP levels were found after exposure to serum from patients with septic shock. The addition of pertussis toxin in the incubation medium constantly increased cAMP response to isoproterenol, but more significantly in PBMC exposed to septic serum. Incubation in the presence of propranolol had no significant effect. The serum of patients with sepsis contained soluble depressant substances that inhibited adenylyl cyclase activation by beta-adrenergic agonists. Septic shock serum exhibited the most potent inhibitory effect. Hyperactivation of the Gi pathway of adenylyl cyclase was mainly responsible for the altered transmembrane beta-adrenergic signaling. PMID- 12578117 TI - Impaired production of interferon-gamma and tumor necrosis factor-alpha but not of interleukin 10 in whole blood of patients with sepsis. AB - It has been demonstrated that lipopolysaccharide (LPS)-induced cytokine response in patients with sepsis differ from the normal host, yet this has not been controlled for the presence of underlying disease. We studied the ability of LPS and killed gram-negative bacteria (GNB) to induce tumor necrosis factor (TNF) alpha and interleukin (IL) 10, and of phytohemagglutinin (PHA) to induce interferon (IFN)-gamma, in whole blood from patients with sepsis (SP, n = 20), patients with matched underlying disease and without sepsis (control patients, n = 20), and healthy volunteers (HV, n = 20). LPS-induced TNF-alpha production was lower in SP (median = 638 pg/mL) compared with control patients (4060 pg/mL; P= 0.003), and control patients production was lower compared with HV (5329 pg/mL; P < 0.001). Pseudomonas aeruginosa-induced TNF-alpha production was lower in SP (1443 pg/mL) than in control patients (7319 pg/mL; P < 0.05), and was not different between control patients and HV (6612 pg/mL; P = 0.6). IFNy production was lower in SP (948 pg/mL) compared with control patients (5516 pg/mL; P < 0.001), and the control patients production was lower compared with HV (11,282 pg/mL; P < 0.001). IL-10 production was not different among the three groups. Down-regulation of TNF-alpha production in patients with sepsis, although not restricted to them, was more pronounced with LPS than with GNB. Although the presence of underlying disease may be involved in the regulatory mechanisms of host response, the use of controls with matched underlying diseases provides strong evidence for the septic condition in the down-regulation of inflammatory response in patients with sepsis. PMID- 12578118 TI - Characterization of the expression of inducible nitric oxide synthase in rat and human liver during hemorrhagic shock. AB - It has been previously shown that the inducible nitric oxide (NO) synthase (iNOS; NOS-2) is elevated after hemorrhage, and that iNOS-derived NO participates in the upregulation of inflammation as well as lung and liver injury postresuscitation from shock. The purpose of this study was to elucidate the time course of iNOS mRNA expression, as well as the cellular and subcellular localization of iNOS protein in the liver posthemorrhage in rats subjected to varying durations of hemorrhagic shock (HS; mean arterial blood pressure [MAP] = 40 mmHg) with or without resuscitation. Expression of iNOS mRNA in rat liver by real-time reverse transcriptase (RT)-PCR demonstrated iNOS upregulation in shocked animals as compared with their sham counterparts as early as 60 min after the initiation of hemorrhage. By 1 h of HS, iNOS protein was detectable in rat liver by immunofluorescence, and this expression increased with time. Immunofluorescence localized iNOS primarily to the hepatocytes, and in particular to hepatocytes in the centrilobular regions. This analysis, confirmed by immunoelectron microscopy, revealed that iNOS colocalizes with catalase, a peroxisomal marker. Furthermore, we determined that iNOS mRNA is detectable by RT-PCR in liver biopsies from human subjects with HS (MAP < 90 mmHg) associated with trauma (n = 18). In contrast, none of the seven nontrauma surgical patients studied had detectable iNOS mRNA in their livers. Collectively, these results suggest that hepatic iNOS expression, associated with peroxisomal localization, is an early molecular response to HS in experimental animals and possibly in human patients with trauma with HS. PMID- 12578119 TI - Effects of cardiac contusion in isolated perfused rat hearts. AB - Myocardial contusion is frequently suspected after blunt chest trauma, but the exact mechanisms of resulting cardiac dysfunction and the time course for enzymatic alterations have not yet been fully understood. Therefore, we investigated pathophysiological aspects of myocardial contusion in a controlled animal model. Male Wistar rat hearts were studied in an isolated perfusion model and were divided into two groups: control (n = 4) and heart contusion (n = 6) groups. The cardiac contusion was produced by a single blow with a weighted pendulum (m = 44 g, height = 20 cm). Functional implications of the contusion were examined in an "isolated perfused heart" model. Troponin 1 concentrations were determined in the perfusate. Cardiac contusion resulted in an increase of coronary perfusion pressure (CPP) of 9 mmHg (P < 0.05, 20 min postcontusion versus baseline and control), followed by a significant increase of left ventricular end-diastolic pressure (LVEDP) of 6 mmHg (P < 0.05, 20 min postcontusion versus baseline and control). Heart contusion was followed by an early increase of troponin 1 (+0.82 ng/mL). The troponin 1 concentration decreased again and, after 20 min, baseline levels were reached. The control group showed no such changes. In this model, high troponin 1 levels after cardiac contusion suggest direct damage to the myocardium. First functional response was shown by the alteration of the coronary perfusion, followed by impaired diastolic function, which persisted even after lowering of the troponin 1 levels. PMID- 12578120 TI - Human dermal microvascular endothelial cells produce nerve growth factor: implications for wound repair. AB - Following cutaneous injury, sensory nerves regenerate into the dermis and epidermis. Tissues that are innervated by sensory nerves synthesize neurotrophins such as nerve growth factor (NGF). The close anatomic proximity of nerves and capillaries throughout the skin suggests that mutual regulation may exist between nerve fibers and microvascular endothelial cells (MECs) during wound repair. Release of the neuropeptide substance P by sensory nerves induces endothelial cell rounding, capillary leak, and cytokine upregulation. We propose that dermal endothelial cells produce neurotrophins required for nerve fiber maintenance and regeneration. In this study, we demonstrate that substance P stimulates NGF messenger RNA expression by cultured human dermal MECs. Likewise, enzyme-linked immunosorbant assay demonstrated that conditioned medium from cultured dermal MECs contains NGF. NGF bioactivity in the supemates was verified by conditioned medium-induced clonal rat pheochromocytoma (PC-12) cell differentiation. This activity was inhibited by anti-NGF antibodies. Therefore, we have demonstrated that substance P, an inflammatory neuropeptide released by sensory nerve fibers, induces endothelial cells to produce NGF. Our data suggest that MECs may be unrecognized contributors to nerve regeneration after cutaneous injury. PMID- 12578122 TI - Burn injury-mediated alterations in cell cycle progression in lymphoid organs of mice. AB - A key event in the cellular and molecular pathogenesis of multiple organ failure (MOF) after burn injury may be the change in profiles of the cell cycle progression in affected organs. We investigated the effects of burn injury on cell cycle progression in immune organs. Cell cycle analysis in the lymphoid tissues of mice after 18% burn injury revealed that S phase entry was temporarily arrested in the thymus 1 day after injury, whereas the spleen had substantially increased S phase entry at day 8. This mode of cell cycle regulation was reproduced in different age groups and strains of mice. Furthermore, the reactivity to the Ki-67 antibody (indicative of proliferation) was markedly reduced in the thymic cortex at day 1. There was a distinct pattern of hematopoietic foci formation and increased reactivities to the Ki-67 antibody in myelogenous cells in the red pulp of spleen at day 7, consistent with the elevated S phase entry. These data suggest that differential regulation of cell cycle progression may play a crucial role in the phenotypic changes in immune organs after burn injury. PMID- 12578121 TI - Flagellin from gram-negative bacteria is a potent mediator of acute pulmonary inflammation in sepsis. AB - Flagellin is a recently identified bacterial product that elicits immune response via toll-like receptor 5. Here, we demonstrate that flagellin is an extraordinarily potent proinflammatory stimulus in the lung during sepsis. In vitro, flagellin triggers the production of interleukin (IL)-8 by human lung epithelial (A549) cells, with 50% of the maximal response obtained at a concentration of 2 x 10(-14) M. Flagellin also induces the expression of ICAM-1 in vitro. Intravenous administration of flagellin to mice elicited a severe acute lung inflammation that was significantly more pronounced than following lipopolysaccharide (LPS) administration. Flagellin induced a local release of proinflammatory cytokines, the accumulation of inflammatory cells, and the development of pulmonary hyperpermeability. These effects were associated with the nuclear translocation of the transcription NF-kappaB in the lung. Flagellin remained active in inducing pulmonary inflammation at doses as low as 10 ng/mouse. In the plasma of patients with sepsis, flagellin levels amounted to 7.1 +/- 0.1 ng/mL. Plasma flagellin levels showed a significant positive correlation with the lung injury score, with the alveolar-arterial oxygen difference as well as with the duration of the sepsis. Flagellin emerges as a potent trigger of acute respiratory complications in gram-negative bacterial sepsis. PMID- 12578123 TI - Natural killer cells participate in bacterial clearance during septic peritonitis through interactions with macrophages. AB - Natural killer (NK) cells have a well-established role in host defense against viral infections and malignancies. However, their function in bacterial infection and sepsis is poorly defined. We hypothesized that NK cells, as a major producer of interferon-gamma during sepsis, would be important in host defense against bacterial infections. Cecal ligation and puncture (CLP) was performed on Swiss Webster mice depleted of NK cells by pretreatment with anti-asialo GM1 and control mice given immunoglobulin G (IgG) antibody. NK cell-depleted mice had significantly higher anaerobic bacterial counts in the liver and peritoneal lavage fluid, as well as higher aerobic counts in the liver and blood 4 h after CLP. Macrophage phagocytosis, nitric oxide production, and interleukin (IL)-6 levels at 4 h were also decreased in mice depleted of NK cells compared with controls. Greater neutrophil influx into the peritoneum, indicated by higher myeloperoxidase levels, was also seen in NK cell-depleted mice. At 8 and 18 h after CLP, bacterial counts were similar between groups, and overall survival rates were not significantly different. Peritoneal IL-12 levels significantly increased by 18 h in normal mice, but not in NK cell-depleted animals. Our data suggest that NK cells participate in the early local and systemic eradication of bacteria and regulation of IL-12 during polymicrobial sepsis. These effects are likely due to their interactions with macrophages. PMID- 12578125 TI - Induction of cytokine tolerance in rodent hepatocytes by chylomicron-bound LPS is low-density lipoprotein receptor dependent. AB - We examined the role of lipoprotein receptors in mediating chylomicron-bound lipopolysaccharide (CM-LPS)-induced cytokine tolerance in rodent hepatocytes. We found that 2 h of pretreatment with CM-LPS (5 mg TG/mL) was sufficient to induce cytokine tolerance, as measured by decreased nitric oxide (NO) production by hepatocytes (20% of the Control group, P < 0.03). Tolerance was evident as early as 2 h after pretreatment and disappeared after 40 h. Furthermore, we evaluated the roles of the low-density lipoprotein (LDL) receptor (LDLR) and LDL receptor related protein (LRP) in the induction of cytokine tolerance in hepatocytes. Biochemical inhibition of the receptors or use of hepatocytes from LDLR-deficient mice revealed that functional LDLR was necessary for the induction of tolerance. However, by increasing the pretreatment time, the LRP compensated for the absence of the LDLR in induction of cytokine tolerance. In conclusion, CM-LPS-mediated induction of cytokine tolerance to proinflammatory cytokines is a time- and dose dependent process that requires functional lipoprotein receptors. These findings underscore an interrelationship between TG-rich lipoprotein metabolism and innate immunity. PMID- 12578124 TI - Differential regulation of cytokines and transcription factors in liver by curcumin following hemorrhage/resuscitation. AB - Inflammatory cytokines interleukin 1 (IL-1), IL-2, IL-6, and tumor necrosis factor-alpha (TNF-alpha) have been recognized as important mediators of pathophysiological and immunological events associated with shock. These inflammatory events after hemorrhage and resuscitation are characterized by the activation of transcription regulators such as nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1). Curcumin, an anti-inflammatory remedy used in Indian medicine, is known to suppress NF-kappaB and AP-1 activation and also to reduce ischemia-reperfusion injuries in animal models. Therefore, the aim of this study was to determine whether administration of curcumin before hemorrhagic shock has any salutary effects on cytokines and the redox-sensitive transcription factors NF-kappaB and AP-1. mRNA levels of IL-1alpha, IL-1beta, IL-2, IL-6, IL 10, and TNF-alpha were determined by reverse transcriptase-polymerase chain reaction in rat livers collected at 2 and 24 h after hemorrhage/resuscitation. The effect of curcumin on the activation of NF-kappaB and AP-1 was determined by electrophoretic mobility shift assays. Significant increases in the levels of liver cytokines IL-1alpha, IL-1beta, IL-2, IL-6, and IL-10 were observed in the 2 h posthemorrhage/resuscitation group compared with sham animals. In contrast, oral administration of curcumin for 7 days followed by hemorrhage/resuscitation regimen resulted in significant restoration of these cytokines to depleted levels, and, in fact, IL-1beta levels were lower than sham levels. Also, the 24-h postresuscitation group showed similar patterns with some exceptions. NF-kappaB and AP-1 were differentially activated at 2 and 24 h posthemorrhage and were inhibited by curcumin pretreatment. Serum aspartate transaminase estimates indicate decreased liver injury in curcumin-pretreated hemorrhage animals. These results suggest that protection against hemorrhage/resuscitation injury by curcumin pretreatment may result from the inactivation of transcription factors involved and regulation of cytokines to beneficial levels. PMID- 12578126 TI - Trypanosoma cruzi sensitizes mice to fulminant SEB-induced shock: overrelease of inflammatory cytokines and independence of Chagas' disease or TCR Vbeta-usage. AB - Trypanosoma cruzi-infected mice display increased susceptibility to shock induced by injection of lipopolysaccharide (LPS), anti-CD3, or resulting from interleukin (IL)-10-defective response to the parasite itself, but the basis of such susceptibility remains unknown. Herein, we tested the susceptibility of mice inoculated with virulent and avirulent T. cruzi to staphylococcal enterotoxins (SE), potent inducers of inflammatory cytokine secretion. Mice infected with T. cruzi CL-strain or inoculated with the avirulent clone CL-14, a clone that does not induce disease or polyclonal lymphocyte activation, succumb suddenly to low doses of staphylococcal enterotoxin B (SEB), but not to staphylococcal enterotoxin A (SEA). High plasma levels of TNF, IFN-gamma, and liver transaminases alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were found in these mice, indicating lethal toxic shock. Sensitization to shock required inoculation of live avirulent trypomastigotes and a time interval before challenge with SEB. We found no prior skewing of T cell receptor (TCR) Vbeta repertoire in CL-14-inoculated mice that could be responsible for sensitization. Splenocytes from CL-14-inoculated mice proliferated more under anti-Vbeta8 than anti-TCRbeta stimulation when compared with normal mice, but were suppressed to SEB stimulation. Both SEB and anti-Vbeta8 antibodies stimulated splenocytes from T. cruzi-inoculated mice to secrete higher levels of inflammatory cytokines than normal controls. Taken together, our results show that T. cruzi inoculation can sensitize mice to lethal SEB-induced shock even in the absence of tissue damage, polyclonal lymphocyte activation, or previously increased levels of inflammatory cytokines, and they suggest that altered reactivity of Vbeta8 lymphocytes may be involved in the phenomenon. PMID- 12578127 TI - Picroliv modulates antioxidant status and down-regulates AP1 transcription factor after hemorrhage and resuscitation. AB - Resuscitation from hemorrhagic shock initiates profound changes in the liver that are likely to contribute to end organ damage and resultant dysfunction after shock. Extensive research in this area has indicated the potential of free radical scavenging strategy for better management of the pathophysiology following hemorrhage-resuscitation (H/R) injury. We studied the effect of a novel pharmacological agent, picroliv, on hepatocellular injury and redox status, as well as its possible mechanism of action in a H/R model in adult rats. Anesthetized rats were subjected to hemorrhagic shock by bleeding 30 mL/kg body weight. After 60 min of shock, rats were resuscitated with twice the shed blood volume of lactated Ringer's solution and were sacrificed 2 h after resuscitation. We observed that picroliv (12 mg/kg) pretreatment, given orally for 7 days, resulted in a significant decrease in serum aspartate transaminase and gamma glutamyl transpeptidase levels. Picroliv also inhibited the lipid peroxidation and nitric oxide release that occurred after H/R and altered the activity of glutathione reductase in a favorable manner, thereby suggesting better antioxidant status. Picroliv significantly down-regulated the stress-sensitive transcription factor AP1 and decreased the level of c-fos mRNA as well as c-jun and c-fos proteins in liver tissue, indicating that its actions could be mediated through AP1 and associated signal transduction pathways. These findings suggest that picroliv has the potential to be developed as a protective agent against H/R injury. PMID- 12578128 TI - Volume expansion with modified hemoglobin solution, colloids, or crystalloid after hemorrhagic shock in rabbits: effects in skeletal muscle oxygen pressure and use versus arterial blood velocity and resistance. AB - Therapeutic goals for hemorrhagic shock resuscitation are the increase of cardiac output and oxygen delivery. The possibility exists that because of microcirculatory effects, different volume expanders result in different tissue oxygen delivery and oxygen use. In a rabbit model of resuscitation from hemorrhagic shock (50% blood loss), we compared the effects of an hemoglobin based O2-carrying solution (HbOC) with those elicited by albumin, hydroxyethyl starch (HES), or saline on systemic hemodynamics, skeletal muscle O2 pressure (PtiO2), and interstitial concentration of lactate (LACi) through the combined implantation of a microdialysis probe and a sensitive O2 electrode into the hind limb. Hemorrhagic shock induced a 50% decrease in mean arterial pressure (MAP), femoral artery blood flow (BF), and PtiO2. After resuscitation, there were statistically significant differences among the volume expanders. The increase in MAP was faster with HbOC and colloids, and slower with saline, mainly obtained by vasoconstriction for HbOC and by increased BF with albumin and HES. The maximum MAP values were significantly higher for HbOC compared with the other volume expanders. HbOC and colloids induced a faster increase in PtiO2 as compared with saline, but maximum PtiO2 values were not different among the volume expanders. Tissue oxygen use as estimated by LACi increased transiently at the beginning of volume expansion with similar maximum values. Animals resuscitated with saline had significantly higher LACi concentrations after the onset of volume expansion as compared with HbOC but not with colloids. Our results demonstrate that there are measurable differences in MAP and BF upon resuscitation with the four different solutions and there is a slower increase in tissue PtiO2 with saline than with colloids associated with significantly increased LACi consistent with delayed reoxygenation upon resuscitation with saline. PMID- 12578129 TI - Dextran modulates microvascular permeability: effect in isotonic and hypertonic solutions. AB - Hypertonic saline solutions with Dextran (HSD) have been advocated for rapid restoration of intravascular volume. Dextran is thought to increase the duration of action of hypertonic saline (HS) by selectively partitioning the water in the vascular space that has been drawn out of cells by HS. The goal of this study was to define the microvascular permeability modulating activity of Dextran in both isotonic and hypertonic solutions. We hypothesized that Dextran would decrease hydraulic permeability (Lp). Using the modified Landis micro-occlusion technique, single rat mesenteric venules were perfused with either normal Ringers (NR) with 135 mM NaCl or HS with 185 mM NaCl. In sequential cannulations of the venules, 1%, 2%, and 3% of Dextran was added to the NR perfusion (n = 6) and the HS perfusion (n = 6). The Lp was measured at baseline and after perfusion with each Dextran concentration. Baseline Lp measurements for NR and HS solutions were 1.01 +/- 0.034 and 5.14 +/- 1.02, respectively. In the NR group, the 2% and 3% Dextran decreased permeability below baseline levels to 0.79 +/- 0.028 (P < 0.0001) and 0.66 +/- 0.028 (P < 0.0001), respectively. In the HS group, the 2% and 3% Dextran decreased permeability to 1.65 +/- 0.53 (P < 0.0001) and 0.99 +/- 0.2 (P < 0.0001), respectively. All values for Lp are x10(-7) cm s(-1) x cm H2O(-1). The addition of Dextran to isotonic and hypertonic solutions results in a decrease in microvessel permeability. This effect is more pronounced with the perfusion of hypertonic solutions. The results demonstrate the oncotic potential of Dextran and its ability to hold water in the vascular space. Dextran may have a beneficial effect when used for resuscitation with HS by decreasing microvascular permeability and augmenting intravascular volume. PMID- 12578131 TI - Sesquiterpene lactone parthenolide, an inhibitor of IkappaB kinase complex and nuclear factor-kappaB, exerts beneficial effects in myocardial reperfusion injury. Shock 17(2):127-134, 2002. PMID- 12578130 TI - Induction of procalcitonin and proinflammatory cytokines in an anhepatic baboon endotoxin shock model. AB - Our objective was to evaluate the role of the liver for procalcitonin (PCT) and cytokine induction in a baboon endotoxin shock model. Complete liver resection with portocaval anastomosis was established in a baboon prior to the induction of endotoxin shock by intravenous administration of endotoxin (100 microg/kg LPS Escherichia coli). Two baboons without surgical intervention were used as controls. Plasma concentrations of PCT, tumor necrosis factor (TNF)-alpha, interleukin (IL) 6, IL-8, endotoxin, and hemodynamic and metabolic parameters were measured pre- and postoperatively and until 6 h after endotoxin administration. PCT concentrations increased to 1.2 and 4.6 ng/mL in control animals at 6 h, but remained below 0.3 ng/mL in the anhepatic baboon. IL-6 and IL 8 increased only for few hours in controls, but remained elevated in the hepatectomized animal near their maximum (IL-6, 2-6 ng/mL) or several-fold higher (IL-8, 30-35 ng/mL), whereas TNF-alpha response was only a small fraction (0.3 ng/mL) of the controls. Endotoxin was much higher and longer persisting in the hepatectomized animal compared with controls. The near absence of PCT production in the anhepatic baboon suggests a primary role for the liver as a source of PCT production during endotoxin shock. Furthermore, the liver also seems to be an important source of TNF-alpha, but not IL-6 or IL-8. PMID- 12578132 TI - Device modeling of ferroelectric memory field-effect transistor for the application of ferroelectric random access memory. AB - An improved theoretical analysis on the electrical characteristics of ferroelectric memory field-effect transistor (FeMFET) is given. First, we propose a new analytical expression for the polarization versus electric field (P-E) for the ferroelectric material. It is determined by one parameter and explicitly includes both the saturated and nonsaturated hysteresis loops. Using this expression, we then examine the operational properties for two practical devices such as the metal-ferroelectric-insulator-semiconductor field-effect transistor (MFIS-FET) and metal-ferroelectric-metal-insulator-semiconductor field-effect transistor (MFMIS-FET) as well. A double integral also has been used, in order to include the possible effects due to the nonuniform field and charge distribution along the channel of the device, to calculate the drain current of FeMFET. By using the relevant material parameters close to the (Bi, La)4Ti3O12 (BLT) system, accurate analyses on the capacitors and FeMFET's at various applied biases are made. We also address the issues of depolarization field and retention time about such a device. PMID- 12578133 TI - A novel beamformer design method for medical ultrasound. Part I: Theory. AB - The design of transmit and receive aperture weightings is a critical step in the development of ultrasound imaging systems. Current design methods are generally iterative, and consequently time consuming and inexact. We describe a new and general ultrasound beamformer design method, the minimum sum squared error (MSSE) technique. The MSSE technique enables aperture design for arbitrary beam patterns (within fundamental limitations imposed by diffraction). It uses a linear algebra formulation to describe the system point spread function (psf) as a function of the aperture weightings. The sum squared error (SSE) between the system psf and the desired or goal psf is minimized, yielding the optimal aperture weightings. We present detailed analysis for continuous wave (CW) and broadband systems. We also discuss several possible applications of the technique, such as the design of aperture weightings that improve the system depth of field, generate limited diffraction transmit beams, and improve the correlation depth of field in translated aperture system geometries. Simulation results are presented in an accompanying paper. PMID- 12578134 TI - A novel beamformer design method for medical ultrasound. Part II: Simulation results. AB - In the first part of this work, we introduced the minimum sum squared error (MSSE) technique of ultrasound beamformer design. This technique enables the optimal design of apertures to achieve arbitrary system responses. In the MSSE technique, aperture weights are calculated and applied to minimize the sum squared error (SSE) between the desired and actual system responses. In this paper, we present the results of simulations performed to illustrate the implementation and validity of the MSSE technique. Continuous wave (CW) and broadband simulations are presented to demonstrate the application of the MSSE method to obtain arbitrary system responses (within fundamental physical limitations of the system). We also describe CW and broadband simulations that implement the MSSE method for improved conventional depth of field (DOF) and for improved correlation DOF in translated aperture geometries. Using the MSSE technique, we improved the conventional DOF by more than 200% in CW simulations and more than 100% in broadband simulations. The correlation DOF in translated aperture geometries was improved by more than 700% in both CW and broadband simulations. PMID- 12578135 TI - Synthetic elevation beamforming and image acquisition capabilities using an 8 x 128 1.75D array. AB - Ultrasound imaging can be improved with higher order arrays through elevation dynamic focusing in future, higher channel count systems. However, modifications to current system hardware could yield increased imaging depth-of-field with 1.75D arrays (arrays with individually addressable elements, several rows in elevation) through the use of synthetic elevation imaging. We describe synthetic elevation beamforming methods and its implementation with our 8 x 128, 1.75D array (Tetrad Co., Englewood, CO). This array has been successfully interfaced with a Siemens Elegra scanner for summed RF and single channel RF data acquisition. Individual rows of the 8 x 128 array can be controlled, allowing for different aperture configurations on transmit and receive beamforming. Advantages of using this array include finer elevation sampling, a larger array footprint for aberration measurements, and elevation focusing. We discuss system tradeoffs that occur in implementing synthetic receive and synthetic transmit/receive elevation focusing and show significant image quality improvements in simulation and phantom data results. PMID- 12578136 TI - A simulation study of sample volume sensitivity for oblique pulsed finite beam insonation of Doppler ultrasound flow phantom cylindrical vessels. AB - Our previous analysis of the lumen pressure in Doppler ultrasound flow phantoms subject to continuous wave, infinite beam excitation is extended here to consider the pressure and Doppler sample volume complex sensitivity within a range of solid absorbent tubes typical of those used in Doppler ultrasound flow phantoms insonated with a focussed pulsed ultrasound beam. The beam may be incident on the cylindrical shell from any angle and with any offset from the shell axis. The examples considered are of a 5 MHz beam with a 6 dB lateral fullwidth of 1 mm at the focus and a transducer surface acceleration pulse with standard deviation of 1 micros propagating through 10 mm outer diameter, 8 mm inner diameter, Cflex, low-density polyethylene (LDPE), high-density polyethylene (HDPE), and polymethylmethacrylate (PMMA) shells surrounded by water at various beam-vessel angles. Our results confirm earlier analyses suggesting that PMMA, being less well matched to the surrounding media, causes much greater distortion of the sample volume sensitivity than Cflex. PMID- 12578137 TI - Differential forms of the Kramers-Kronig dispersion relations. AB - Differential forms of the Kramers-Kronig dispersion relations provide an alternative to the integral Kramers-Kronig dispersion relations for comparison with finite-bandwidth experimental data. The differential forms of the Kramers Kronig relations are developed in the context of tempered distributions. Results are illustrated for media with attenuation obeying an arbitrary frequency power law (alpha(omega) = alpha0 + alpha1(absolute value of omega)y). Dispersion predictions using the differential dispersion relations are compared to the measured dispersion for a series of specimens (two polymers, an egg yolk, and two liquids) exhibiting attenuation obeying a frequency power law (1.00 < or = y < or = 1.99), with very good agreement found. For this form of ultrasonic attenuation, the differential Kramers-Kronig dispersion prediction is found to be identical to the (integral) Kramers-Kronig dispersion prediction. PMID- 12578138 TI - The highly sensitive optical measurement of absolute SAW amplitudes for power flow analysis. AB - The established optical method for the measurement of surface acoustic wave (SAW) amplitudes (simple probing) has been modified to result in the effective separation of scattered light from the SAW-diffracted light by means of two single mode optical fibers. In this way, the photodetector noise is drastically reduced, and absolute SAW amplitudes of 10(-12) m can be detected easily. Using an additional precise measurement of the nondiffracted light by means of fiber shifting, the SAW amplitudes can be calibrated with an error of less than 3%. This method can be applied to deliver an accurate power flow analysis in SAW devices. As an example, for a YZ-LiNbO3 transducer, the measured and calculated SAW power densities were found to agree within better than 6%. PMID- 12578139 TI - Plate equations for piezoelectrically actuated flexural mode ultrasound transducers. AB - This paper considers variational methods to derive two-dimensional plate equations for piezoelectrically actuated flexural mode ultrasound transducers. In the absence of analytical expressions for the equivalent circuit parameters of a flexural mode transducer, it is difficult to calculate its optimal parameters and dimensions, and to choose suitable materials. The influence of coupling between flexural and extensional deformation, and coupling between the structure and the acoustic volume on the dynamic response of piezoelectrically actuated flexural mode transducer is analyzed using variational methods. Variational methods are applied to derive two-dimensional plate equations for the transducer, and to calculate the coupled electromechanical field variables. In these methods, the variations across the thickness direction vanish by using the stress resultants. Thus, two-dimensional plate equations for a stepwise laminated circular plate are obtained. PMID- 12578141 TI - SAW filters including one-focus slanted finger interdigital transducers. AB - The present paper describes a new filter type, slanted finger interdigital transducer (SFIT) that allows fast analysis as a precondition for fast optimization. Therefore, filter structure and analysis are especially suitable for optimizing unidirectional SFIT (USFIT) filters. For analysis, a SFIT filter is usually divided into many narrow channels. Every channel is considered to be a subfilter and is analyzed separately. Therefore, the total filter analysis is very time consuming. For reducing computing time, we suggest a one-focus structure. In contrast to conventional SFIT filters, not only the finger and gap widths but also the spaces between transducers of different channels differ by one and the same scaling factor. As a consequence, all prolonged finger edges of both transducers intersect in one point called focus. As a result, the parameters of all subfilters can be calculated from the parameters of only one subfilter by simple frequency scaling. Consequently, the total time for analysis is essentially reduced. However, one-focus SFIT filters with continuous finger edges show a deep minimum within the passband. This problem can be overcome by using stepped one-focus structures. PMID- 12578140 TI - Velocity fluctuation reduction in vector Doppler ultrasound using a hybrid single/dual-beam algorithm. AB - In order to reduce the fluctuations in the velocity magnitude estimate, we propose a modification to the standard algorithm for reconstructing the (two component) vector velocity from the measured Doppler shifts in two directions. This uses the standard dual-beam algorithm, combined with temporal smoothing, to find only the velocity angle, then uses the single-beam algorithm to estimate the velocity magnitude. We present initial data showing the significant reduction in velocity estimate fluctuation that this hybrid method achieves compared to the standard algorithm. PMID- 12578142 TI - Influence of silicon dioxide-silicon interface trap charges on the performance of monolithic metal-zinc oxide-silicon nitride-silicon dioxide-silicon convolver. AB - An enhancement in the convolution efficiency is obtained by annihilating the SiO2 Si interface trap charges in the metal-ZnO-Si3N4-SiO2-Si convolver structure. The annealing process uses a source of hydrogen created underneath the SiO2-Si interface by implanting H3(+) ion followed by rapid thermal anneal of 5s at 900 degrees C. The silicon nitride layer is inducted to protect ZnO films from hydrogen influx during low temperature oxygen anneal. PMID- 12578143 TI - Notes on synthetic phased-array with 2-D sparse arrays. AB - Recent advances in 3-dimensional (3-D) imaging with 2-D sparse arrays have generated renewed interest in array synthesis theory. The 3-D imaging with arbitrary planar line shapes is particularly interesting. In this correspondence, we clarify some points presented in recent publications and show that aperture synthesis can be viewed with a very simple, physical approach. Its limitations due to approximations also will be discussed. PMID- 12578144 TI - Are current rapid detection tests for Group A Streptococci sensitive enough? Evaluation of 2 commercial kits. AB - A new, 1-step, enzyme-linked immunoassay kit for detection of Group A Streptococci (GAS) in throat samples (QuickVue In-Line One-Step Strep A Test; Quidel Corporation, San Diego, CA) was evaluated for use in a study comprising 536 patients in 8 primary healthcare centres. Compared to conventional culture at the clinical microbiology laboratory, the sensitivity achieved was 73.9% and the specificity 86.8%; these figures were not affected to any major extent by broth enrichment of samples before culturing or following PCR testing of the cysteine proteinase gene for independent diagnosis of GAS. It was also found that most samples containing low numbers of GAS were missed by the rapid test. We therefore evaluated the kit in use in our area (TestPack Plus Strep A Test; Abbott Laboratories, Chicago, IL) in a separate study of 615 patients. Somewhat increased sensitivity (82.8%) and specificity (96.1%) were obtained. As current antigen tests depend on subjective judgement of test outcome, improvements in test design or provision of more detailed instructions may be desirable in order to achieve optimal results. PMID- 12578145 TI - Ochrobactrum anthropi bacteraemia in immunocompetent children. AB - Ochrobactrum anthropi is an emerging pathogen in immunocompromised patients but infections with the bacterium have very rarely been documented in normal hosts. We report the characteristics of O. anthropi bacteraemia in 11 immunocompetent children, aged 2 months to 7 y, hospitalized in a general hospital during a 5-y period. Children commonly presented with fever, non-specific respiratory or gastrointestinal manifestations, leukocytosis and neutrophilia and had a rapid recovery, even when they did not receive a specific treatment. In 10 cases positive blood cultures were obtained on admission and in all cases subsequent cultures were sterile. In conclusion, O. anthropi may cause bacteraemia in immunocompetent hosts, although further studies are required to clarify whether these isolates represent pseudobacteraemia or whether O. anthropi is a potential pathogen of low virulence. PMID- 12578146 TI - Molecular characteristics of Neisseria meningitidis strains isolated in Burkina Faso in 2001. AB - In the course of an epidemic of meningitis in Burkina Faso in 2001, 27 cerebrospinal fluid samples from patients in 7 districts were forwarded to Norway for isolation and characterization of the causative agents. Neisseria meningitidis was isolated from 13 (48%) samples. The isolates were analysed using serological and genetic methods. Of the 13 strains, 4 were serogroup A, serotype 21:P1.9, sequence type (ST)-5 and belonged to clonal subgroup III, while the remaining 9 strains were serogroup W135, serotype 2a:P1.5,2, ST-11 and belonged to the electrophoretic type-37 complex. PCR analyses revealed meningococcal DNA in 13/14 culture-negative samples. Sequence analysis of the PCR products demonstrated that at least 3 different meningococcal strains were responsible for these 13 cases. Our results show that the W135 strain associated with the 2000 hajj (Muslim pilgrimage) outbreak was a significant cause of disease in Burkina Faso in 2001. Further studies are warranted to determine whether W135 is about to replace serogroup A in sub-Saharan Africa. PMID- 12578147 TI - Is acute hepatitis B still a nosocomially-acquired infection in Poland? A case controlled study of transmission routes in a hospital sample. AB - The aim of this study was to evaluate the role of medically linked risk factors for acute HBV infection in the Podlasie region of North-Eastern Poland. The records of 107 adult patients and 214 age- and sex-matched controls hospitalized in the Department of Infectious Diseases, Bialystok, Podlasie Region during the period 1997-2000 with the diagnosis of acute hepatitis B were reviewed. On multivariate analysis, household contact with hepatitis B [odds ratio (OR) = 18.0, 95% confidence interval (CI) 2.0-165.5] and a history of injections (OR = 2.7, 95% CI 1.5-4.7) in the preceding 6 months were independently associated with acute HBV infection. In conclusion, our data indicate that despite a significant decline in the incidence of hepatitis B, injections are still involved in the spread of HBV in Poland. PMID- 12578148 TI - Tuberculous meningitis in a country with a low incidence of tuberculosis: still a serious disease and a diagnostic challenge. AB - In order to assess the present epidemiology, clinical presentation and outcome of patients with tuberculous meningitis (Tm), a retrospective study was conducted including 20 Tm patients admitted to a referral department in Denmark between January 1988 and July 2000. The findings were compared to those of a similar survey conducted 12 y earlier. A total of 65% of the patients came from countries with a high endemicity of tuberculosis, compared to only 26% in the previous survey. The overall mortality rate was 20%, with elderly patients with various pre-existing diseases being particularly affected. Neurological sequelae were seen in 50% of patients and seemed to be related to the duration of symptoms and treatment delay. The neurological state on admission and a rapid progression of symptoms seemed to be related to mortality. In 85% of the patients treatment was initiated without a definitive bacteriological diagnosis having been reached. The findings indicate that Tm is a rare but still serious disease, and one that is associated with high mortality and morbidity. PMID- 12578149 TI - Intestinal Candida phospholipase is not elevated in patients with antibiotic associated diarrhea. AB - In order to assess the role of Candida-secreted phospholipase in antibiotic associated diarrhea (AAD), 43 fecal Candida isolates from patients with AAD and from controls were tested on egg yolk agar for production of phospholipase. Phospholipase zones did not differ between the isolates from patients with AAD and from controls. The data indicate that the fungal virulence factor phospholipase may not be responsible for AAD in adults. PMID- 12578150 TI - Late complications of Candida (Torulopsis) glabrata fungemia: description of a phenomenon. AB - Complications occurring > 1 month after Candida glabrata fungemia have not been studied. We conducted a study to determine the frequency and morbidity of complications of C. glabrata bloodstream infections occurring > 1 month after infection. Late complications were common (incidence 9.9 per 100 patient years), and most often occurred within the first year. In a multivariate analysis, late complications were associated with the presence of diabetes mellitus [odds ratio (OR) 12.2; 95% confidence interval (CI) 1.2-130] and chronic renal failure (OR 14.7; 95% CI 1.2-184). These data suggest that careful long-term follow-up of patients with C. glabrata fungemia is important and present an opportunity to explore secondary prophylaxis. PMID- 12578151 TI - Invasive pulmonary aspergillosis: identification of risk factors. AB - Aspergillosis is the second most frequent fungal infection after candidiasis in teaching hospitals. Clinical manifestations of pulmonary aspergillosis range from asymptomatic colonization to disseminated disease. The aim of this study was to identify the risk factors associated with invasive pulmonary aspergillosis, in patients with positive pulmonary isolation of Aspergillus species. A review was undertaken of all clinical records with pulmonary isolation of Aspergillus species at Reina Sofia University Hospital from January 1995 to December 1998. Data collected were: age, gender, history of smoking, past medical history, such as chronic pulmonary disease, immunosuppression, granulocytopenia in the past 6 months and during the last admission, history of surgery including within the last year of the study period, number of hospital admissions and clinical evidence of invasive pulmonary aspergillosis. To investigate all the possible risk factors for invasive pulmonary aspergillosis, a multivariable logistic regression model was used. 132 patients with positive pulmonary isolation were identified, of which 42.4% had clinical evidence of invasive pulmonary aspergillosis. The independent factors significantly associated with invasive pulmonary aspergillosis were: granulocytopenia in the past 6 months, immunosuppression in the last admission and the number of hospital admissions within the past year. Patients with a history of granulocytopenia in the past 6 months and immunosuppression in the last admission are the high-risk group for invasive pulmonary aspergillosis. However, invasive pulmonary aspergillosis can also occur in mild granulocytopenic or even immunocompetent patients. PMID- 12578152 TI - Lack of effect of food on the bioavailability of a new ketolide antibacterial, telithromycin. AB - Telithromycin is an innovative antibacterial designed for the treatment of community-acquired respiratory tract infections. This study assessed the effect of food on the bioavailability of a single oral dose of telithromycin 800 mg in healthy male subjects. Male volunteers aged 18-45 y were recruited for an open label, single-dose, 2-period, cross-over study. In each trial period, subjects received a single oral dose of telithromycin 800 mg after an overnight fast, or after a standard high-fat breakfast. A washout period of 6-8 d separated the 2 study periods. All 18 subjects recruited (mean age 30.7 y) completed the study. Telithromycin was rapidly absorbed, reaching maximum plasma concentrations within a median of 2.50 and 2.25 h in the fasting and non-fasting states, respectively. There was no statistical difference between the non-fasting and fasting states for any of the pharmacokinetic parameters measured. The mean plasma telithromycin concentration versus time profiles for the non-fasting and fasting phases were almost superimposable. For the maximum plasma concentration and area under the curve from time 0 to infinity, the 90% confidence intervals for the mean non fasting:fasting ratios were 83-116 and 101-123 mg x h/l, respectively; these are within 80-125% of the bioequivalence range. Telithromycin was well tolerated. The bioavailability, rate and extent of absorption of the new ketolide antibacterial telithromycin were unaffected by food. PMID- 12578153 TI - Diagnostic tools in respiratory tract infections: use and comparison with Finnish guidelines. AB - The objectives of this prospective epidemiological study were to describe the diagnosis and treatment of respiratory tract infections by Finnish general practitioners and to compare current practice with national evidence-based guidelines. All patients (n = 4386) seeking primary care for a respiratory tract infection for the first time in 30 health centres during 1 week in November 1998 participated in the study. The main outcome measures were the amounts and types of diagnostic tests used and antimicrobials prescribed. Tympanometry was used in 1% of patients with acute otitis media. Ultrasonography, sinus radiography or both were used in 80% of cases of sinusitis and antigen detection or culture for Streptococci in 57% of throat infections. In acute bronchitis, a chest radiograph was taken in 5% of cases and the CRP level determined in 8%. The corresponding figures for pneumonia were 49% and 39%. In pneumonia and throat infection, diagnostic testing was statistically significantly associated with the use of antimicrobials, but not in otitis, sinusitis or acute bronchitis. Diagnostic tests were underused in respiratory tract infections compared to evidence-based recommendations. PMID- 12578154 TI - ICU-acquired nosocomial infection: impact of delay of adequate antibiotic treatment. AB - In order to measure the impact on survival of the early introduction of adequate antibiotic treatment for nosocomial bacteremia and pneumonia, a retrospective, cohort study was carried out over a period of 17 months in a 6-bed respiratory ICU. All patients presenting with a first episode of ICU-acquired nosocomial bacteremic infection (Centers for Disease Control criteria) or pneumonia [BAL culture > or = 10(4) colony-forming units (CFU)/ml or protected specimen brush culture > or = 10(3) CFU/ml] were included. The organ failure score (Fagon criteria) was recorded on the day of diagnosis. Adequate antibiotic treatment was defined by the sensitivity of each etiologic organism to at least 1 prescribed antibiotic. A total of 25 patients (Simplified Acute Physiology Score II = 44) were included in the study with pneumonia (n = 17) or bacteremia (n = 8), on average 6.5 +/- 4.6 d after admission. At the time of diagnosis, 23 patients were receiving mechanical ventilation. The overall mortality rate was 48% and was significantly associated with the length of time without adequate antibiotic treatment (p = 0.011) and the number of organ failures on the day of diagnosis (p = 0.017). Adequate antibiotic treatment only had an impact on survival if it was started within the first 24 h after sampling (p < 0.02 on Day 0 and < 0.04 on Day 1). On the day of diagnosis, a failure score > 2 was associated with increased mortality (p = 0.009). After adjusting for the number of organ failures, the length of time without adequate antibiotic treatment remained associated with mortality (< or = 2 organ failures, p < 0.02; > 2 organ failures, p = 0.05). This study suggests that, during the course of nosocomial pneumonia and bacteremia, the time at which adequate antibiotic treatment is started is a key factor influencing survival. PMID- 12578155 TI - Transient hypophosphataemia associated with acute infectious disease in paediatric patients. AB - The aim of this study was to investigate the prevalence of hypophosphataemia in children with acute infection and the relationship between serum phosphate and C reactive protein (CRP) concentration. Serum phosphate and CRP levels were measured on admission in 238 patients (aged 1 month to 14 y) with: pneumonia (n = 51), upper respiratory tract-related bacterial infection (n = 70), urinary tract infection (n = 50) and viral infection (n = 67). Patients were classified according to CRP value (0-50, 51-100, 101-150, > or = 151 mg/l) and type of infection. The prevalence of hypophosphataemia was calculated for each group. 30 children with hypophosphataemia on admission had serial measurements of serum phosphate and CRP levels. A significant negative correlation between serum phosphate and CRP levels was found (r = -0.41, p < 0.0001). Patients with CRP > or = 151 mg/l on admission had a lower mean serum phosphate value than those with CRP < or = 50 mg/l (1.17 vs 1.50 mmol/l, p < 0.0001). The overall prevalence of hypophosphataemia for patients with pneumonia, upper respiratory tract bacterial infection, urinary tract and viral infections was 45%, 35.7%, 18% and 4.4%, respectively. Hypophosphataemia occurred during the phase of rising of CRP level and resolved soon after CRP reached a plateau. In conclusion, hypophosphataemia is a relatively frequent but transient phenomenon in children with acute infectious disease. It is associated with an increase in CRP concentration and resolves before the normalization of CRP levels. PMID- 12578156 TI - Risk of acquiring tick bites in south-eastern Sweden. AB - The incidence and the temporal pattern of tick bites were studied in a population frequently out-of-doors in a tick-endemic area in south-eastern Sweden between May 2000 and March 2001. The participants, who were well aware of tick-borne diseases, inspected their skin daily from May until September and completed a diary sheet, registering visited geographical places, time out-of-doors, observed tick bites, etc. The participants were also given questionnaires in both the initial and final stages of the study, asking questions about their earlier history of tick bites, previous tick-borne diseases and out-of-doors activities. The incidence was 0.04 [95% confidence interval (95% CI) 0.02-0.061 tick-bitten participants/10 h spent out-of-doors. In total, the participants registered 1767 tick bites, i.e. an incidence of 0.14 (95% CI 0.10-0.18) tick bites/10 h out-of doors. Within 6 months after the registration period, 8/235 (3%) had been treated for physician-diagnosed Lyme borreliosis. In conclusion, this study found a 4% risk of being tick-bitten per 10 h spent out-of-doors. The risk of contracting Lyme borreliosis was 1/221 tick bites (0.5%, 95% CI 0.44-0.56). Thus, the results indicate a low risk of acquiring Lyme borreliosis when using daily tick checks and we underline the opinion of not recommending routine prophylactic treatment for observed tick bites. PMID- 12578157 TI - Livedo reticularis associated with the use of a midline catheter. AB - We report a case of livedo reticularis as an unusual complication of a midline catheter in a patient being treated for pyelonephritis with intravenous antibiotics. The rash resolved completely after catheter removal. The constellation of symptoms suggested an aberrant autonomic response as the cause of the illness. PMID- 12578158 TI - Primary group A streptococcal peritonitis in a previously healthy child. AB - Primary peritonitis in a child without underlying medical conditions is rare outside the neonatal period. A girl with no past medical history presented with acute abdominal pain. Laparotomy revealed primary peritonitis due to group A Streptococcus (serotype emm89). She was treated with antibiotics and immune globulin, and recovered fully. PMID- 12578159 TI - Psoas abscess diagnosed at a Northern university hospital. AB - Abscess of the psoas muscle is an infrequent diagnosis at hospitals in Northern countries. We report on 16 patients who had this diagnosis during the period 1991 2001. Eight patients were immigrants who had previously been healthy and most of them had experienced symptoms for approximately 1 y. MRI or CT scans revealed spondylodiscitis in 6 of these patients and Mycobacterium tuberculosis was identified as the causative agent. With the exception of 1 patient who was exclusively treated with antituberculous agents, all 8 immigrant patients were successfully treated with antituberculous agents in addition to percutaneous drainage. The other 8 patients were Norwegians, 4 of whom had underlying conditions such as diabetes mellitus or drug abuse. The causative microorganisms were Staphylococcus aureus or Streptococcus spp., with the exception of M. tuberculosis in 1 case. The Norwegian patients had a more acute history of symptoms than the immigrant patients and 2 of them were in a septic condition on admittance. Two of the Norwegians died of serious infection; 5 were successfully treated with percutaneous drainage in addition to antibiotics and 1 was treated exclusively with antibiotic agents. The clinical history and microorganism associated with psoas abscess seemed to depend on whether or not the patient was an immigrant. Owing to increasing immigration, diagnosis of psoas abscess should be taken into account in Northern countries. PMID- 12578160 TI - Moraxella catarrhalis bacteraemia associated with Mycoplasma pneumoniae infection and pneumonia. AB - This report describes a case of severe pneumonia associated with Mycoplasma pneumoniae infection and Moraxella catarrhalis bacteraemia in a 44-y-old woman with undiagnosed breast carcinoma. M. pneumoniae is increasingly recognized as a co-pathogen but to the authors' knowledge this is the first reported case of M. catarrhalis bacteraemia associated with M. pneumoniae infection. PMID- 12578161 TI - Treatment of human granulocytic ehrlichiosis during pregnancy and risk of perinatal transmission. AB - A pregnant woman from Nantucket Island, MA was diagnosed with human granulocytic ehrlichiosis at 34 weeks gestation. We describe the diagnostic and therapeutic dilemmas involved and discuss the risks of perinatal transmission. PMID- 12578162 TI - Strongyloides stercoralis infection as a cause of acute granulomatous appendicitis in an HIV-positive patient in Athens, Greece. AB - A case of acute granulomatous appendicitis due to Strongyloides stercoralis infection in an HIV-positive patient is described. To our knowledge this is the first case presented in the literature. PMID- 12578163 TI - Cryptococcus neoformans myositis without dissemination. AB - We report a case of isolated cryptococcal myositis involving the paraspinal muscle without evidence of disseminated disease in a patient with a large B-cell lymphoma diagnosed at the time of presentation. Biopsy of the muscle involved grew a pure culture of Cryptococcus neoformans and periodic acid-Schiff staining showed numerous budding yeast consistent with Cryptococcus spp. The patient responded to systemic antifungal therapy with complete resolution of his infection. We also present a review of 5 previously published cases of cryptococcal myositis. PMID- 12578164 TI - Recurrent isosporiasis over a decade in an immunocompetent host successfully treated with pyrimethamine. AB - Recurrent isosporiasis in an immunocompetent host is reported. The patient suffered from chronic intermittent diarrhea for over a decade. Multiple short term administrations of trimethoprim-sulfamethoxazole followed by pyrimethamine, or albendazole combined with tinidazole could not control the relapses. However, treatment with pyrimethamine, 25 mg/d for 20 weeks, was successful. PMID- 12578165 TI - Cerebral mycotic aneurysm complicating Stomatococcus mucilaginosus infective endocarditis. AB - Stomatococcus mucilaginosus is a gram-positive coccus present in the normal flora of the mouth and upper respiratory tract of humans. Although traditionally believed to be an organism of low virulence, S. mucilaginosus has recently been recognized as an emerging opportunistic pathogen, especially in patients with chronic immunosuppressive diseases. This report describes the case of a patient with known mitral valve prolapse, who presented with a mycotic cerebral aneurysm revealing a non-nosocomial spontaneous S. mucilaginosus endocarditis. The spectrum of infections due to this opportunistic pathogen is reviewed, with particular focus on assessing the clinical characteristics and prognosis of S. mucilaginosus infective endocarditis. PMID- 12578166 TI - Membrane bioreactors for wastewater treatment and reuse: a success story. AB - This paper briefly reviews our current understanding of membrane bioreactor processes (including conventional and immersed membrane designs) resulting from three decades of development, highlighting process advantages, applications and areas for continuing research. PMID- 12578167 TI - Japanese experiences of environmental management. AB - Japan experienced a very rapid industrialization and economic growth in the era of income doubling in 1960s and at the same time Japan experienced very severe damage from various types of environmental pollution. In this paper, historical development of population, GNP, energy consumption with classification of petroleum, coal and electric power, and CO2 emission are introduced as basic background data on Japanese development. The tragic experience of Minamata disease and Itai-itai disease caused by methyl mercury and cadmium, respectively, are introduced. In two tables, historical development of water pollution and air pollution are summarized. Regarding solid wastes management, the total mass balance in Japan and recent development in legislation framework for enhancement of recycling of wastes are introduced briefly. PMID- 12578168 TI - Integrated centralized utility services to a chemical complex on Jurong Island, Singapore. AB - SUT pioneered centralized utility services for the chemical industry on Jurong Island, which are cost-effective due to economies of scale, reliable due to inter connection of satellite operations, and customer tailored for special requirements. The utility services range from the supply of steam and water, wastewater treatment, incineration, terminalling, service corridor to fire fighting. Among the services, water management achieves the complete cycle from wastewater treatment to effluent recycling. PMID- 12578169 TI - Findings of the Beijing 2000 IWA Chemical Industries Speciality Conference. AB - Critical Technologies to the World in the 21st Century: Pollution Control and Reclamation in Process Industries was a Chemical Industries Specialty Conference held in Beijing, China 18-22 September 2000. Set in China, the cradle of civilization, the conference hosted about 100 delegates from over twenty countries. Approximately 122 technical papers were presented covering a wide range of topic areas. This paper reviews significant findings presented. Perhaps the conference proceedings will serve as a point of reference for future evaluations and accomplishments. The 21st century offers both challenges and opportunities for progress towards sustainable development/production in the process industries. International cooperation and inter-disciplinary collaboration are essential for this goal to be realized. PMID- 12578171 TI - A pesticide runoff model for simulating runoff losses of pesticides from agricultural lands. AB - An integrated modeling system was developed to predict runoff losses of pesticides from agricultural lands. The system is an integration of a mathematical model, a database system, and a geographic information system. Information on soil type, land use, land slope, watershed boundaries, precipitation, pesticide usage, as well as physical and chemical properties of pesticides have been input to a GIS, managed through a database, and used for further modeling studies. The modeling outputs were in turn put into the database, such that runoff patterns along with pesticides losses could be further simulated by using a database management system. The final results could then be visualized through GIS. The developed modeling system was applied to the Kintore Creek Watershed, Ontario, Canada, for simulating losses of atrazine from agricultural lands. A water quality monitoring project was carried out from 1988 to 1992 in the watershed to detect conditions of surface water pollution due to the use of pesticides. The modeling outputs were verified through the monitoring data, demonstrating reasonable prediction accuracy. The result indicated that the model provides an effective means for forecasting pesticide runoff from agriculture lands. PMID- 12578170 TI - Magnetic separation of iron and heavy metals from water. AB - A magnetic separation device is being developed for removal of iron and heavy metals from water. The device consists of a column of supported magnetite surrounded by a movable permanent magnet. The mineral magnetite, or synthetically prepared iron ferrite (FeO x Fe2O3), is typically supported on various materials to permit adequate water passage through the column. In the presence of an external magnetic field, enhanced capacity was observed in using supported magnetite for removal of actinides and heavy metals from wastewater. The enhanced capacity is primarily due to magnetic filtration of colloidal and nanoscale particles along with some complex and ion exchange sorption mechanisms. This paper will review some previous work on the use of magnetite for wastewater treatment and discuss the development and potential of the magnetic nanoscale filtration/sorption process for water treatment. Recent research results are also presented on preliminary experimental studies using the process with water samples containing iron. PMID- 12578172 TI - Coagulation of humic acid by ferric chloride in saline (marine) water conditions. AB - The coagulation of a model seawater-humic acid solution with a hydrolysis metal salt (FeCl3) has been studied by monitoring floc size, solution pH, and zeta potential. The kinetic features of the orthokinetic coagulation have been demonstrated in relation to coagulant dosages, solution pH and zeta potential. Humic acid removal and floc charge reduction increased with coagulant dosage. Adjusting the solution pH prior to coagulation had a substantial effect on the treatment performance. By pH adjustment to pH 6, the greatest humic acid removal (by coagulation and subsequent membrane filtration) and the largest floc size was achieved at a FeCl3 dosage of 200 micromol l(-1). It is believed that the coagulation is characterised by competition between OH- ions and humic acid for ferric ions in the co-precipitation process. In acidic pH, where the concentration of OH- ions is low, humic acid molecules may compete more favourably for bonding sites in the co-precipitation, which leads to a more compact precipitation and a higher overall humic acid removal. PMID- 12578173 TI - Effect of dissolved oxygen concentration on the biofilm and in situ analysis by fluorescence in situ hybridization (FISH) and microelectrodes. AB - A better understanding of microbiology and ecology of nitrifying bacteria in inner biofilms is an important part of improving process performance and control. Microelectrodes and fluorescent in situ hybridization (FISH) in biofilm research have been used to investigate the spatial distributions of various microbial activities in biofilms and have led to new experimental findings as well as modifications of the homogeneous assumptions in the biofilm kinetic models. The objective of this study is to try the combination of two methods, both FISH and microelectrode measurements, and to provide reliable and in situ information on nitrifying bacterial activity in biofilms. The characteristics of biofilm developed on tygon slides were different according to the change of dissolved oxygen (DO). When the DO increased from 2 to 10 mg DO/L, the rate of the biofilm thickness increased and its dry density changed from 50-70 to 25-90 mg/cm3. Ammonia oxidizing bacteria were not uniformly distributed in biofilm, and were found at the deeper layer where oxygen is depleted, they were detected primarily in the upper and middle layers of the biofilm. PMID- 12578174 TI - Wastewater reuse in on-site wastewater treatment: bacteria and virus movement in unsaturated flow through sand filter. AB - In on-site wastewater treatment plants, effluents are pre-treated by septic tank and treated by soil infiltration or sand filtration systems, with unsaturated flow conditions. These systems remove efficiently carbon, nitrogen and suspended solids. But for microbial pollution, the treatment efficiency depends on the hydrodynamic behaviour and filtering media characteristics. Contamination of superficial water and groundwater due to pathogenic viruses and pathogenic bacteria is responsible for many diseases. The objective of this study is to approach the mechanisms and operating conditions to control bacteria and virus release in the environment. Experiments were carried out on reactors of different length packed with sand. Hydraulic load of 90 cm x d(-1) with a pulse periodic flow was used. The influence of chemical composition of the solution on the treatment efficiency has also been studied. For the first time, the residence time distribution (RTD) has been studied using a conservative tracer (KI), to determine the main hydrodynamic parameters. For the second time, the RTD with bacterial and viral tracers (E. coli, bacteriophage MS2) was applied, with the aim to define microbial behaviour in filtering media, including adsorption and filtration phenomena. This work allowed us to determine retardation factors according to the hydraulic loads and chemical composition. PMID- 12578175 TI - Effects of salt concentration on floc characteristics and pollutants removal efficiencies in treatment of seafood wastewater by SBR. AB - Most seafood wastewater has been treated by the activated sludge process. Due to changes in salt concentration, pollutant loadings and raw materials, the process is not operated satisfactorily to meet effluent regulation. Most problems faced at present are solid liquid separation. In this study, effects of salt concentration on floc characteristics and pollutants removal efficiencies were investigated in treatment of seafood wastewater by SBR. For analyzing fractal dimension of flocs, the small angle laser light scattering (SALLS) method was applied using a Diffraction Particle Sizer (Malvern Instruments). Organic removal efficiencies (in terms of COD(Mn)) decreased with increasing salt concentration, but eventually reached a steady state. Fractal dimension and floc size also showed similar trends with changing salt concentration. The main reasons to reduced pollutant removal efficiencies were deteriorated biological activity and settling properties. The biological activity was affected faster than the floc characteristics by increasing salt concentration. The deteriorated settling properties were explained by decreased size and fractal dimension of floc due to increasing salt concentration. The settling properties of floc such as sludge volume index (SVI) and zone settling velocity (ZSV) were related to size and fractal dimension of floc. The fractal dimension of floc was better related to the settling properties than the size of floc. PMID- 12578176 TI - Nitrogen removal using a vertically moving biofilm system. AB - The aim of this laboratory study was to establish the efficacy of a new experimental biofilm system for the removal of nitrogen from synthetic wastewater. The system consisted of six reactors in series: one anaerobic, one anoxic and four aerobic reactors. In both the anaerobic and anoxic reactors, a plastic cuboid module was repeatedly moved up and down in the wastewater, while being totally submerged at all times. In each of the aerobic reactors, an identical module to that used in the anaerobic and anoxic reactors was intermittently and repeatedly immersed in and lifted out of the wastewater. All the individual reactors had a bulk fluid volume of 28.2 litres and the average temperature of the wastewater was about 10 degrees C. Each module consisted of crossflow corrugated plastic sheets with a surface area of 1.824 m2. The nitrate recycle flow from the fourth aerobic tank to the anoxic tank was twice the inflow to that tank. In the anoxic reactor, filtered COD was removed at an average rate of 2.22 kg COD/m3 x d and nitrate-nitrogen was denitrified at a rate of 0.42 kg NO3-N/m3 x d. The average nitrification rate in the second aerobic reactor was 0.12 kg NH4-N/m3 x d. The new biofilm system was simple to construct and operate. PMID- 12578177 TI - Catalytic ozonation of 2-dichlorophenol by metallic ions. AB - This research is mainly to explore functional improvement by adding various kinds of metallic ions in the ozonation of 2-chlorophenol solution. During the experiment, various kinds of metallic ions (Pb+, Cu2+, Zn2+, Fe2+, Ti2+, and Mn2+) were added; it was found that the reaction rate increased in all cases. The best result was obtained by using manganese ions, followed by iron and titanium ions. At pH = 3 and 1 ppm Mn2+ concentration, the reaction rate was increased by three times. TOC removal rate was also increased from 12.6% to 38% at 60 min reaction time. Ozone self-decomposition with various kinds of metallic ions alone was tested. It was found the ozone self-decomposition coefficient is highly dependent on the reaction rate constant for ozonation of 2-chlorophenol. The improvement of reaction was relevant to the mechanism of reaction between ozone and metallic ions. Furthermore, the effect of adding manganese ions was studied. With the initial manganese concentration at 0-2 ppm, after gas exposure for 20 min the removal rate can be increased from 38% to 93%. TOC removal rate was increased from 11% to 38%. The reaction rate was improved more greatly at the initial pH = 3. PMID- 12578178 TI - Adsorption characteristics of benzene on biosolid adsorbent. AB - This study selected biosolids from a petrochemical wastewater treatment plant as the raw material. The sludge was immersed in 0 to 5 mol l(-1) of ZnCl2 solutions and pyrolyzed at different temperatures. When the sludge was pyrolyzed for 30 min at temperatures of 400, 500, 600, and 700 degrees C, the corresponding surface area of the biosolid adsorbent was 46, 401, 921, and 727 m2/g, respectively. Pore size analysis indicated that the mesopore (20 to 500 A) contributed more than the macropore and micropore in the sludge pyrolytic residue. When the benzene influent concentration was 800 ppmv, the adsorption capacity ranged from 59 to 164 mg/g for different biosolid adsorbents. A larger BET surface area and smaller average pore diameter yielded a larger benzene adsorption capacity. PMID- 12578179 TI - Characteristics of coagulation-flocculation of humic acid with effective performance of polymeric flocculant and inorganic coagulant. AB - Ferric chloride and aluminium sulphate as coagulants and positive charged flocculants PDDMAC ((PDDMAC = poly (diallyldimethylammonium chloride) were used for pre-treatment of water and wastewater for removing humic substance prior to RO membrane filtration. It was found that a combination of flocculant and coagulant enhanced the coagulation-flocculation process and humic acid removal. The optimum conditions of coagulation-flocculation were established in reference to the ratio of humic acid and coagulant. Zeta potential and the ratio of E4/E6 were investigated to explore the possible micro-mechanisms of coagulation flocculation. The ratios of E4/E6 show the molecular size variations using different coagulants and flocculants, which are expected to benefit membrane fouling control. PMID- 12578180 TI - Nitrifying microbial community analysis of nitrite accumulating biofilm reactor by fluorescence in situ hybridization. AB - Biological nitrogen removal via nitrite pathway in wastewater treatment is very important especially in the cost of aeration and as an electron donor for denitrification. Wastewater nitrification and nitrite accumulations were carried out in a biofllm reactor. The biofilm reactor showed almost complete nitrification and most of the oxidized ammonium was present as nitrite at the ammonium load of 1.2 kg N/m3/d. Nitrite accumulation was achieved by the selective inhibition of nitrite oxidizers by free ammonia and oxygen limitation. Nitrite oxidation activity was recovered as soon as the inhibition factor was removed. Fluorescence in situ hybridization studies of the nitrite accumulating biofilm system have shown that genus Nitrosomonas which is specifically hybridized with probe NSM156 was the dominant nitrifying bacteria while Nitrospira was less abundant than those of normal nitrification systems. Further FISH analysis showed that the combinations of Nitrosomonas and Nitrospira cells were identified as important populations of nitrifying bacteria in an autotrophic nitrifying biofilm system. PMID- 12578181 TI - A submerged tubular ceramic membrane bioreactor for high strength wastewater treatment. AB - A 4 L submerged tubular ceramic membrane bioreactor (MBR) was applied in laboratory scale to treat 2,400 mg-COD/L high strength wastewater. A prolonged sludge retention time (SRT) of 200 day, in contrast to the conventional SRT of 5 to 15 days, was explored in this study, aiming to reduce substantially the amount of disposed sludge. The MBR system was operated for a period of 142 days in four runs, differentiated by specific oxygen utilization rate (SOUR) and hydraulic retention time (HRT). It was found that the MBR system produced more than 99% of suspended solid reduction. Mixed liquor suspended solids (MLSS) was found to be adversely proportional to HRT, and in general higher than the value from a conventional wastewater treatment plant. A chemical oxygen demand (COD) removal efficiency was achieved as high as 98% in Run 1, when SOUR was in the range of 100-200 mg-O/g-MLVSS/hr. Unexpectedly, the COD removal efficiency in Run 2 to 4 was higher than 92%, on average, where higher HRT and abnormally low SOUR of 20 30 mg-O/g-MLVSS/hr prevailed. It was noted that the ceramic membrane presented a significant soluble nutrient rejection when the microbial metabolism of biological treatment broke down. PMID- 12578182 TI - Granular activated carbon (GAC) adsorption in tertiary wastewater treatment: experiments and models. AB - Wastewater treatment has always been a major concern in the developed countries. Over the last few decades, activated carbon adsorption has gained importance as an alternative tertiary wastewater treatment and purification process. In this study, granular activated carbon (GAC) adsorption was evaluated in terms of total organic carbon (TOC) removal from low strength synthetic wastewater. This paper provides details on adsorption experiments conducted on synthetic wastewater to develop suitable adsorption isotherms. Although the inorganics used in the synthetic wastewater solution had an overall unfavourable effect on adsorption of organics, the GAC adsorption system was found to be effective in removing TOC from the wastewater. This study showed that equation of state (EOS) theory was able to fit the adsorption isotherm results more precisely than the most commonly used Freundlich isotherm. Biodegradation of the organics with time was the most crucial and important aspect of the system and it was taken into account in determining the isotherm parameters. Initial organic concentration of the wastewater was the determining factor of the model parameters, and hence the isotherm parameters were determined covering a wide range of initial organic concentrations of the wastewater. As such, the isotherm parameters derived using the EOS theory could predict the batch adsorption and fixed bed adsorption results of the multi-component system successfully. The isotherm parameters showed a significant effect on the determination of the mass transfer coefficients in batch and fixed bed systems. PMID- 12578183 TI - Operational factors of submerged inorganic membrane bioreactor for organic wastewater treatment: sludge concentration and aeration rate. AB - Various sludge concentrations and aeration rates were evaluated to find the optimal operation condition of a submerged ceramic membrane filtration system. 5.6 g/L of sludge was diluted with water to concentrations of 2.8 g/L and 1.4 g/L, and the three sludge concentrations were compared in terms of titration characteristics such as pressure and filtrate flux. Flux was at the highest value of about 30 L/m2 x hr at 50 kPa when sludge concentration was 1.4 g/L. In contrast, when sludge concentrations increased to 2.8 g/L and 5.6 g/L, the flux at 50 kPa decreased significantly to 18 L/m2 x hr and 10 L/m2 x hr, respectively. It was concluded that the sludge concentration directly affected the filtration efficiency, and low sludge concentration was suitable for improving filtration efficiency. Adjusting the aeration rate from 2 L/min to 4 L/min at 5.6 g/L of sludge and 50 kPa of pressure increased flux from 10 L/m2 x hr to 13 L/m2 x hr. It was obvious that the vigorous aeration improved the filtration efficiency, but the aeration rate did not seem to be high enough to maintain flux lower than critical flux. PMID- 12578184 TI - Evaluation of polyaluminium ferric chloride (PAFC) as a composite coagulant for water and wastewater treatment. AB - Coal gangue is a kind of waste from coal mine processing. Polyaluminium ferric chloride (PAFC), a new type of inorganic composite coagulant, was prepared by using the waste from the Mineral Bureau of Yanzhou, China, hydrochloric acid and calcium carbonate as raw materials. The relationship between the stability of ferric ion and the ionic strength of solution was investigated. The zeta potential of PAFC hydrolysis products of PAFC and the coagulation performances under different pH value were discussed. The turbidity removal properties of PAFC, polyaluminium (PAC) and polyferric sulfate (PFS) were compared, and the color removal effect of PAFC for the wastewater containing suspended dyes was also tested. In addition, the coagulation performance of PAFC for actual wastewaters from petrochemical plant, iron and steel plant, and coal mining processing was evaluated. The experimental results suggest that PAFC took a maximum value of zeta potential at about pH 5.8 on the positive side. Compared with PAC, PAFC gives better turbidity removal performance in the range of pH from 7.0 to 8.4. PAFC gives good color removal performance on suspension dyes. PAFC also gives good wastewater purifying results for the actual wastewater. Therefore, PAFC is a high-effect and stable water treatment agent. PMID- 12578185 TI - Non-woven fabric filter separation activated sludge reactor for domestic wastewater reclamation. AB - A non-woven fabric filter was experimentally evaluated for solid-liquid separation in an activated sludge reactor as an alternative membrane. A polypropylene fabric filter (70, 50 and 35 g/m2) was used for the experiment. The pilot system was operated in A/O (Anaerobic/Oxic) type in which the filter module was submerged into the oxic compartment. The filtration module consists of 10 plate type rectangular filter elements with effective filtration area, 2 m2. Gravity filtration was carried out for solid-liquid separation by changing the water head 0.05-0.5 m without backwashing during the system operation. Initial permeate flux was set at 0.4 m/d. C/N ratio of raw wastewater was controlled at 4.5 in terms of BOD/T-N. The fabric filter system showed a good performance enough for domestic wastewater treatment. Effluent solid concentration was 3.2 mg/L (93.5% removal). COD removal efficiency was 91.6% producing an effluent concentration around 13 mg/L. 66% of total nitrogen removal could be obtained at the adjusted C/N ratio of influent wastewater. However phosphorus removal was very low at 23%. It was found that the initial flux of 0.4 m/d should be maintained for stable performance of the system. PMID- 12578186 TI - Performance and membrane fouling in a pilot scale SBR process coupled with membrane. AB - The performance of the pilot-scale submerged membrane coupled with sequencing batch reactor (SM-SBR) for upgrading effluent quality was investigated in this study. The reactor was operated with 3-hour cycle with alternating anoxic and aerobic conditions to treat organics, nitrogen and phosphate. Despite various influent characteristics, COD removal was always higher than 95%. Sufficient nitrification was obtained within a few weeks after start-up and during the stable period, complete nitrification occurred despite short aeration time. Total nitrogen (TN) removal efficiency was reached up to 85%. Membrane flux was critical for TN removal so that the decrease of flux by membrane fouling led to increase of HRT, and it caused the endogenous respiration of microorganisms such as nitrifying bacteria. The stirred cell test revealed the significant role of the soluble fraction in membrane permeability and dissolved solids played a major role in the short-term fouling mechanism. The cake resistance by the soluble COD fraction of supernatant or soluble microbial products (SMP) was investigated as a major part of total resistance. PMID- 12578187 TI - Coagulation of humic-kaolin-PACI aggregates. AB - Water treatment residual flocs are fractal-like aggregates made of many initial aggregates. We investigated in this study the coagulation dynamics for the humic mineral-polyaluminium chloride (PACI) aggregates using small-angle light scattering techniques and the free-settling test. In contrast to reports in the literature, the presence of humic acid did not lead to a loose floc. Not only the time evolution of the coagulation dynamics, but also the final floc characteristics are only mildly affected by the humic acid. However, the strength of the formed floc does decline with humic acid, which leads to a turbid supernatant with high level of organics. PMID- 12578188 TI - Hydrogen production from wastewater by acidogenic granular sludge. AB - Sludge was granulated in a hydrogen-producing acidogenic reactor when operated at 26 degrees C, pH 5.5 treating a sucrose-rich wastewater. The influence of hydraulic retention time (HRT) and sucrose concentration on hydrogen production by the acidogenic granular sludge was investigated at a constant loading rate of 25 g-sucrose/(l x day). Results show that the gas composition was not greatly influenced by HRT or sucrose concentration. The hydrogen accounted for 57% to 68% of the biogas at HRT ranging 4.6-28.6 h and sucrose concentration ranging 4,800 29,800 mg/l. However, the hydrogen yield was more dependent on HRT and sucrose concentration. It ranged from 0.19 to 0.27 l/g-sucrose with the maximum yield occurring at HRT 13.7 h and sucrose concentration 14,300 mg/l in the wastewater. The acidified effluent was composed of volatile fatty acids and alcohols. The predominant products were butyrate (59-68%) and acetate (10-25%), plus smaller amounts of i-butyrate, valerate, i-valerate, caproate, methanol, ethanol, propanol, and butanol. The sludge yield averaged 0.2 g-VSS/g-sucrose. The carbon balance was 98-107% throughout the study. PMID- 12578189 TI - Nutrient removal using anaerobically fermented leachate of food waste in the BNR process. AB - Nutrients removal efficiencies highly depend on the presence of biodegradable organic carbon in the biological nutrient removal (BNR) process but most domestic wastewater in Korea has shown a low C/N ratio and has a small amount of biodegradable COD (chemical oxygen demand). On the other hand, about 11,577 tons of food waste that contains a lot of organic material has been produced in Korea per day. The feasibility and applicability of anaerobically fermented leachate of food waste (AFLFW) as an external carbon source was examined in the laboratory scale BNR process at different operation conditions with synthetic wastewater and domestic sewage. As the addition of AFLFW increased, the average removal efficiencies of SCOD, T-N, T-P changed from 96%, 60%, and 2% to 90%, 77%, and 67%, respectively. From anoxic nitrate utilization tests, it was observed that once the readily biodegradable COD (especially VFAs) was depleted, the denitrification rate reduced from 8.2 mg NO3-N/g VSS/hr to 0.7 mg NO3-N/g VSS/hr. From the molecular size distribution test, it was concluded that about 60% of soluble COD in effluent, which was considered to originate from AFLFW, had a large molecular size (> 30 kDa) that was not used by microorganisms. PMID- 12578190 TI - Extraction of Cu and Pb from printed circuit board sludge using ammonia solutions. AB - Sludge from a printed circuit board factory containing high concentrations of Cu and Pb was characterized. Aqueous ammonia solutions were used to extract metals from the sludge. The extraction reactions were completed within 6 hrs. The best extraction efficiency was found at pH of 10.0. Higher solid-liquid ratio and higher ammonia concentration resulted in better extraction efficiency. Fractionation experiments showed that Cu and Pb were mainly extracted from the Fe Mn oxide-bound and carbonate-bound fractions. Extracted sludge could meet the TCLP regulation limit and be categorized as a non-hazardous waste. Results show that ammonia extraction is of potential in resource recovery and in detoxification of hazardous waste. PMID- 12578191 TI - Manufacturing artificial aggregates from industrial sludge and marine clay with addition of sodium salt. AB - The potential of converting industrial sludge and dredged marine clay into building and construction materials as an alternative to disposal was investigated in this study. The industrial sludge was mixed with marine clay at various compositions and was shaped into round pellets to be used as concrete aggregates. The pellets were then dried and transferred into a high temperature kiln where they were heated at 1,135 degrees C. The artificial aggregates were more porous then the normal granite aggregate. The occurrence of this condition was suspected to be attributed to two factors, viz. the firing temperature and the amount of sodium in the aggregates. To validate this hypothesis, the aggregates with and without added sodium chloride were fired at temperatures of 1,100 and 1, 200 degrees C. Results showed a reduction in the densities of the aggregates with the addition of sodium chloride and the increase in firing temperature. PMID- 12578192 TI - The reaction mechanism of catalytic oxidation with hydrogen peroxide and ozone in aqueous solution. AB - The sorption and catalytic oxidation of model compounds (pCBA and phenanthrene) and NOM on FeOOH were investigated using hydrogen peroxide and ozone, respectively. After oxidation with ozone, the hydrophobic, transphilic, and hydrophilic NOM fractions were isolated using XAD-8 and -4 resins to analyze the reaction characteristics. The sorption of pCBA was strongly dependent upon the pH, but phenanthrene exhibited a sorption behavior that was independent of the pH. In the case of NOM, the hydrophobic portion showed higher sorption affinity than hydrophilic and transphilic at pH 7.2. The concentrations of model compounds and oxidants were measured during the oxidations and the efficiency was compared for tests done with ozone alone and those using catalytic ozonation. Through the comparison of the sorption and decomposition of the model compounds, along with the effects of bicarbonate addition, mechanisms for catalytic oxidation with hydrogen peroxide or ozone were proposed, respectively. PMID- 12578193 TI - Adsorption of heavy metals from water using banana and orange peels. AB - Liquid-phase adsorption removal of Cu2+, Co2+, Ni2+, Zn2+, and Pb2+ in the concentration range of 5-25 mg/L using low-cost banana and orange peel wastes was examined at 30 degrees C. Under comparable conditions, the amount of adsorption decreased in the order Pb2+ > Ni2+ > Zn2+ > Cu2+ > Co2+ for both adsorbents. The adsorption isotherms could be better described by the Freundlich equation. The amount of adsorption increased with increasing pH and reached a plateau at pH > 7, which was confirmed by the variations of zeta potentials. The application potential of such cellulose-based wastes for metal removal (up to 7.97 mg Pb2+ per gram of banana peel at pH 5.5) at trace levels appeared to be promising. PMID- 12578194 TI - Destruction of model organic pollutants in water using ozone, UV and their combination (Part I). AB - The experimental results showed that ozone (O3) oxidation is an effective means to destroy phenolic organic pollutants present in water. High removal efficiencies can be readily achieved for most of the model compounds fortified in aqueous matrices within a reasonable time frame. This study also included the oxidation of phenolic compounds using ozone in combination with ultraviolet (UV) irradiation. The extent of mineralization measured in terms of the losses in total organic carbon is promoted by the joint action of ozone and UV in comparison with using ozone alone. PMID- 12578195 TI - Observations of blanket characteristics in full-scale floc blanket clarifiers. AB - We monitored the changes in concentrations, zeta potentials, sizes and capillary suction times of the solids flocs in the clarified water from eight floc blanket clarifiers of PingTsan Water Works of Taiwan Water Supply Company with low (< 10 NTU) and high (> 100 NTU) turbidity raw water. For the former, one-stage coagulation-sedimentation treatment was adopted which yielded a rather unstable blanket. Complete washout was noticeable when the PACl dosage was insufficient. On the treatment of high-turbidity raw water, on the other hand, the Works adopted the combined treatment process, that is, the raw water was first coagulated and settled in a pre-sedimentation tank, afterwards, its effluent was coagulated again and clarified in the clarifiers. The resulting flocs could form a networked blanket that was relatively stable to the shock load in raw water turbidity. PMID- 12578196 TI - Dissolved air flotation clarification of activated sludge and wastewaters from chemical industry. AB - Wastewaters from separate chemical factories are treated together in an extended aeration activated sludge plant. The factories produce chemicals for paper industry (e.g. starch), latexes and animal feed. The components of the wastewaters include styrene, tertiary butanol and vinyl acetate. Activated sludge is clarified by sedimentation. During winter time, when the water temperature was 3-12 degrees C, the clarification deteriorated causing carry over of suspended solids containing COD. Enhancement of suspended solids and COD removals was studied in a dissolved air flotation jar test unit. Flotation trials were conducted for activated sludge, sedimentation treated final effluent (tertiary treatment) and separate wastewater fractions. The need for chemicals, flocculation and amount of recycle water were judged according to the achieved removals. Dissolved air flotation was found well suited for the clarification of activated sludge, but not technically and economically feasible for the clarification of the wastewater streams before the activated sludge treatment. PMID- 12578197 TI - Photocatalytic oxidation technology for humic acid removal using a nano structured TiO2/Fe2O3 catalyst. AB - A novel TiO2 coated haematite photocatalyst was prepared and used for removal of colored humic acids from wastewater in an UV bubble photocatalytic reactor. XRD analysis confirmed that nano-size anatase crystals of TiO2 were formed after calcination at 480 degrees C. SEM results revealed that nano-size particles of TiO2 were uniformly coated on the surface of Fe2O3 to form a bulk of nano structured photocatalyst Fe2O3/TiO2. The porous catalyst had a BET surface area of 168 m2/g. Both the color and total organic carbon (TOC) conversion versus the residence time were measured at various conditions. The effects of pH value, catalyst loaded, initial humic acid concentration and reaction temperature on conversion were monitored. The experimental results proved that the photocatalytic oxidation process was not temperature sensitive and the optimum catalyst loading was found to be 0.4 g/l. Degradation and decolorization of humic acids have higher efficiency in acidic medium and at low initial humic acid concentration. The new catalyst was effective in removing TOC at 61.58% and color400 at 93.25% at 180 minutes illumination time and for 20 mg/l neutral humic acid aqueous solution. The kinetic analysis showed thatthe rate of photocatalytic degradation of humic acids obeyed the first order reaction kinetics. PMID- 12578198 TI - Tertiary nitrification in a new dual-medium aerated filter. AB - In an effort to develop an aerated filter with high nitrification capabilities, we formulated a new filter with 2 layers: the upper layer is filled with a plastic medium, and the lower with anthracite. We tested the filter in a pilot scale apparatus on secondary-treated sewage, and in a small-scale apparatus on secondary-treated sewage and on artificial sewage. We evaluated the nitrification characteristics under various conditions of water velocity, air velocity, temperature, and pH. We designed a tertiary plant to treat 20,000 m3/day of secondary-treated sewage, using the new filter or a conventional filter. From these comparisons, it appeared that the dual-medium aerated filter had high nitrification capabilities and was an effective device for tertiary sewage treatment. PMID- 12578199 TI - Competitive adsorption for binary mixture of 4-nitrophenol and phenol on RSTA using GAC. AB - Adsorption characteristics of a reverse stratified tapered adsorber (RSTA) system and competitive adsorption for phenol and 4-nitrophenol were investigated. In this study, a novel technology, RSTA, was introduced and compared with a conventional cylindrical adsorber (CA). The optimum study of RSTA in a single solute system with phenol for size distribution, mean velocity and optimum angle was conducted to satisfy environmental criteria and reduce granular activated carbon (GAC) treatment costs. The competitive adsorption efficiency of RSTA and CA was also compared in a binary solute system with phenol and 4-nitrophenol. In the results of single-solute experiment, breakthrough time of RSTA was increased by 30-50% over conventional CA. The mean bed velocity was 19.10 cm/min and the optimum angle of RSTA was decided to 3 degrees. Adsorption efficiency was increased with increasing GAC dose and the number of GAC layers. The affinity of competitive adsorption was significantly increased with decreasing GAC dose and increasing adsorbate concentration. The RSTA can be considered to provide an increase in breakthrough time with decreasing flow rate and increasing the number of layers. Water and wastewater plants processing large volumes of water can realize significant savings in capital and operation costs by utilizing this type of operation. PMID- 12578200 TI - Nitrate removal by simultaneous sulfur utilizing autotrophic and heterotrophic denitrification under different organics and alkalinity conditions: batch experiments. AB - The effect of various organic compounds were tested using lab-scale batch reactors. At sufficient alkalinity, the initial nitrate nitrogen concentration of 100 mg/L was completely reduced in all batch reactors. Sulfate production decreased by the addition of organics. The concentration range of organics used in this experiment did not inhibit autotrophic denitrification except for propionate. Propionate inhibited autotrophic denitrification a little, indicated by a lower sulfate production rate. Biomass in suspension increased with higher initial organic concentrations, showing higher DOC consumption. As the concentration of organics increased, alkalinity increased accordingly. Under the conditions of low alkalinity, in the case of a control reactor without organics, only about 30% of the initial nitrate was reduced. With half the theoretically required dosage of methanol, the denitrification rates increased slightly. When ethanol, acetate, and propionate were used, denitrification went to completion. When excess organics was added, however, sulfate production was significantly decreased. Interestingly, even when small amounts of organics were added, autotrophic denitrification was promoted as indicated by the sulfate production. PMID- 12578201 TI - Features of double chamber bed and case study. AB - A double chamber bed (DCB) was developed in China in 1980 and has been utilised in more than 500 setups. This paper introduces the structure and operation features of DCB, and also the unique structure of DCB leads to its perfect operation features. It can be also considered as the best option in reconstruction projects which can replace other types of ion exchangers, such as fixed bed, double cell fluidized-bed, multi-bed. Four reconstruction project cases were introduced, which were selected from more than twenty plants designed by us. Through the case study, it is concluded that DCB can not only be used to treat different sources of water (river and well), but can also be used to treat water with the ratio of transient hardness to total cation to be less than 0.5 and the ratio of strong to weak anion to be more than 7.0. These findings enlarge the scope of application of DCB. PMID- 12578202 TI - Bacterial biosorbent for removing and recovering copper from electroplating effluents. AB - Investigations were carried out to study the removal and recovery of Cu(II) ions from wastewater by Micrococcus sp. The Langmuir isotherm model described very well the equilibrium behavior of copper biosorption, with maximum biosorption capacity (q(max)) reaching 52.1 mg Cu2+/g dry cell at pH 6. Biomass prewashed with sulfuric acid (0.05 mol 1(-1)) and sodium sulfate (1 mol l(-1)) solutions were shown to increase the copper removal capabilities up to 27% and 16%, respectively. Copper uptake by cells was negligible at pH 2.0 and then increased quickly with increasing pH until 6.0. Cells of Micrococcus sp. were immobilized in 2% calcium alginate and 10% polyacrylamide gel beads. A counter-current process comprising a series of immobilized cell reactors was developed for removing and recovering copper from electroplating effluents. This process was capable of producing an effluent at low copper concentration, with only a minimum amount of desorbing agent used. The technique of scanning electron microscopy coupled with X-ray dispersion analysis shows that Cu2+ exchanged with K+ and Ca2+ on the cell wall of Micrococcus sp., thereby suggesting ion exchange as one of the dominant mechanisms of metal biosorption for this bacterial strain. PMID- 12578203 TI - A simplified risk-based approach for process screening in municipal wastewater reclamation and reuse. AB - Reuse of municipal wastewater has become a promising solution to relieve the tension of increasing fresh water demand in many metropolitans. Although different advanced technologies are available to reclaim wastewater into premium quality, associated health effects are usually not properly assessed in reclamation process selection. A simplified risk-based approach developed for process screening and adaptation of health impacts as a consideration in reclamation process selection is discussed in this paper. This approach can be used to screen out unqualified processes and those with poor cost benefits. As a result, the design of wastewater reclamation could be enhanced to control the associated health impacts in wastewater reuse. PMID- 12578204 TI - Fouling and cleaning of microfiltration membrane in municipal wastewater reclamation. AB - This paper presents an investigation on fouling and cleaning of a tubular microfiltration membrane for municipal wastewater reclamation. A bi-level complex model, namely channel clogging and pore plugging, was introduced to elucidate the fouling mechanism. Based on 135 days of microfiltration of activated sludge, a preventive method for channel clogging was reasonably proposed and proven to be effective. Without channel clogging, the operation period was observed a five fold increase on average, reaching 3-4 weeks. To remove pore plugging, a multi step chemical cleaning was employed and further optimized in terms of temperature and alkaline concentration. Generally, the chemical cleaning could restore the membrane permeability to higher than 90%. PMID- 12578205 TI - Biological treatment of printing ink wastewater. AB - Printing ink wastewater is usually very difficult to treat biologically and its chemical oxygen demand (COD) far exceeds standards of discharge. The COD in wastewater is usually 3,000 to 8,000 mg/L after flocculation and sedimentation. Herein, a strain of bacterium was isolated from the sludge and identified as Bacillus sp. and utilized to treat printing ink wastewater. The application of bacteria to degrade printing ink in wastewater is discussed in this paper. The influence of N and P sources on COD removal, and COD removal in combination with glucose was also discussed. More than 85 per cent of the COD could be removed using the proposed biological process. A novel internal airlift loop bioreactor with bacteria immobilized onto ceramic honeycomb support was used for the wastewater treatment. PMID- 12578206 TI - Classification of lake sediments using a hydrocyclone. AB - The present work is a preliminary experiment for hydrocyclones as a primary process in lake sediment thickening. A few cyclones based on the Rietema standard geometry were prepared, and sample particles were sediments from a local lake and waste coal fly ash for a reference test. As a result of the chemical analysis, more organic contaminants were found in smaller particles. The experimental tests showed that physical characteristics of particles, configuration of the cyclone and operation conditions could affect the separation efficiency. PMID- 12578207 TI - A culture-independent approach for studying microbial diversity in aerobic granules. AB - This study reports the use of ribosomal-based molecular techniques to study the microbial diversity in aerobic granules. Aerobic granules at different growth stages (young, mature and old) were obtained from a laboratory scale sequential aerobic sludge blanket (SASB) bioreactor fed with glucose as the main source of carbon and energy. Deoxyribonucleic acid (DNA) was extracted from the young, mature and old granules. The polymerase chain reaction (PCR) was used to amplify the Eubacterial 16S ribosomal DNA (rDNA) and three clone libraries were constructed, corresponding to each of the three growth stages. The microbial diversity in each clone library was assessed by amplified ribosomal DNA restriction analysis (ARDRA). The results reveal that there was considerable diversity in each clone library and there were variations in microbial diversity among the three different clone libraries. This suggests a shift in the composition of the microbial communities. Microorganisms associated with 5 restriction fragment length polymorphism (RFLP) types (A, B, C, D and E) appear to play an important role in the development of aerobic granules. PMID- 12578208 TI - The performance of a two-stage SBR system in treating dye containing wastewater. AB - A two-stage SBR system treating the wastewater containing copper-phthalocyanin dye-Reactive Turquoise Blue KN-G (C. I. Reactive Blue 21, denoted by RTB) was investigated during a 200-cycle operation. The performance of the system, including pollutant removal rates, operating stability and sludge characteristics, may be a concern in the long-term run. The results shows that the system removed RTB efficiently despite the step-up RTB concentration from 13.1 to 107 mg/L in the influent. The average total removal rates of RTB were 81% to 92.5% due to the contribution of both anaerobic and aerobic stages, while stable effluent was produced with the help of the aerobic stage. The sludge in each reactor was in the steady state and of good activity on RTB removal. Moreover, the anaerobic sludge with the SVI value of 109.1 and the aerobic sludge with the SVI value of 103.2 had good settling properties, which was verified by hardly any presence of suspended solids in the effluent and an observation under an electron-scanning microscope. The adsorption and biodegradation were considered as the mechanism for the stability of the SBR system during the long term run. PMID- 12578209 TI - Setting-up a control simulation strategy for a sequencing batch reactor (SBR): application to municipal wastewater. AB - The use of a simulation model for setting up a control strategy for a sequencing batch reactor necessary for treating municipal wastewater is described. The model used is the ASM no 1 model. The objective of the pollution control treatment is the removal of carbon and nitrogen; the optimisation is concerned with the improvement in the biological removal of nitrogen. After experimental identification of the initialisation variables, the model enables different SBR control scenarios to be tested (time variation for each process) leading to the total elimination of nitrogen. The best simulation was tested in a laboratory reactor. On that scale, it was noted that denitrification is an endogenous process. Lastly, the control strategy was tested on a semi-industrial pilot working in a pollution control plant. Other control scenarios can be devised and tested by simulation, in order to improve the productivity of the reactor. PMID- 12578210 TI - Isolation of naphthalene-degrading bacteria from tropical marine sediments. AB - Oil pollution is a major environmental concern in many countries, and this has led to a concerted effort in studying the feasibility of using oil-degrading bacteria for bioremediation. Although many oil-degrading bacteria have been isolated from different environments, environmental conditions can impose a selection pressure on the types of bacteria that can reside in a particular environment. This study reports the successful isolation of two indigenous naphthalene-degrading bacteria from oil-contaminated tropical marine sediments by enrichment culture. Strains MN-005 and MN-006 were characterized using an extensive range of biochemical tests. The 16S ribosomal deoxyribonucleic acid (rDNA) sequence analysis was also performed for the two strains. Their naphthalene degradation capabilities were determined using gas chromatography and DAPI counting of bacterial cells. Strains MN-005 and MN-006 are phenotypically and phylogenetically different from each other, and belong to the genera Staphylococcus and Micrococcus, respectively. Strains MN-005 and MN-006 had maximal specific growth rates (micro(max)) of 0.082 +/- 0.008 and 0.30 +/- 0.02 per hour, respectively, and half-saturation constants (K(s)) of 0.79 +/- 0.10 and 2.52 +/- 0.32 mg per litre, respectively. These physiological and growth studies are useful in assessing the potential of these indigenous isolates for in situ or ex situ naphthalene pollutant bioremediation in tropical marine environments. PMID- 12578211 TI - A GIS-based modeling system for petroleum waste management. Geographical information system. AB - With an urgent need for effective management of petroleum-contaminated sites, a GIS-aided simulation (GISSIM) system is presented in this study. The GISSIM contains two components: an advanced 3D numerical model and a geographical information system (GIS), which are integrated within a general framework. The modeling component undertakes simulation for the fate of contaminants in subsurface unsaturated and saturated zones. The GIS component is used in three areas throughout the system development and implementation process: (i) managing spatial and non-spatial databases; (ii) linking inputs, model, and outputs; and (iii) providing an interface between the GISSIM and its users. The developed system is applied to a North American case study. Concentrations of benzene, toluene, and xylenes in groundwater under a petroleum-contaminated site are dynamically simulated. Reasonable outputs have been obtained and presented graphically. They provide quantitative and scientific bases for further assessment of site-contamination impacts and risks, as well as decisions on practical remediation actions. PMID- 12578212 TI - A comparative study of anaerobic digestion of food waste in a single pass, a leachate recycle and coupled solid/liquid reactors. AB - A single pass reactor (R1), a leachate recycle reactor (R2) and a coupled solid/liquid bioreactor (R3-Rm) for anaerobic digestion of food waste were comparatively investigated in terms of digestion process and treatment efficiency. The coupled solid/liquid bioreactor is an enhanced two-phase system and distinctive from a traditional two-phase process with an upflow anaerobic sludge blanket (UASB) reactor as the methanogenic phase and a circulation of treated leachate between the acidification and methanogenic phases. In comparison with R1 and R2, R3-Rm enhanced the digestion process and increased the methane content of biogas. 100% of the R3-Rm methane yield was from the methanogenic phase with average methane content of 71%. The significant enhancement was also confirmed by the removal of 79% of total organic carbon (TOC), 60% of volatile solids (VS) and 80% of total COD in 12 days running of R3-Rm. However, no active methane fermentation was detected in R1 and R2 during 60 days operation. The results in this laboratory-scale study show that the rapid accumulation of volatile fatty acids (VFAs) due to the rapid acidification of food waste inhibits the development of effective methane fermentation in single pass and leachate recycle reactors. The coupled solid/liquid bioreactor is more efficient in converting food waste into methane and carbon dioxide. PMID- 12578213 TI - Investigations on the wear behaviour of the temporary PMMA-based hip Spacer-G. AB - Total hip replacement has become one of the most successful orthopaedic procedures. However, complications due to infections may give serious problems and have devastating consequences for the hip implant. The use of a temporary three-dimensional polymethylmethacrylate (PMMA) cement spacer may be an alternative to solve infections in hip implants, improving the lives of patients awaiting reimplantation. In order to evaluate their wear behaviour, five PMMA Spacer-G femoral heads were tested against five post-mortem pelves in a hip joint simulator with bovine calf serum as lubricant. The surface of the worn spacers was characterized by scanning electron microscopy (SEM) analysis; all the samples revealed a similar morphology, showing areas characterized by different degrees of wear. Particle debris was isolated from the lubricant and PMMA particles and bone fractions were quantified. The amount of debris was found to be higher than where no-temporary prostheses were used. However, this result is acceptable since wear debris is removed by lavage irrigation when the Spacer-G is explanted. On the basis of these data, it is considered that the use of the cement Spacer-G could be a promising approach to the treatment of complicated infections of the hip joint. Therefore, Spacer-G is worthy of further research. PMID- 12578214 TI - Fatigue strength of PMMA bone cement mixed with gentamicin and barium sulphate vs pure PMMA. AB - Barium sulphate is added to polymethylmethacrylate (PMMA) bone cement as a radiopacifier. Gentamicin is an antibiotic added to bone cement to treat or prevent infection in arthroplasty. This study investigated the combined effect of barium sulphate and gentamicin sulphate on the fatigue strength of PMMA bone cement. Three different formulations were studied: pure PMMA, PMMA with barium sulphate added and PMMA with barium sulphate and gentamicin sulphate added. Before testing all specimens were stored in water at 37 degrees C for at least 15 days to season the PMMA and to elute the antibiotic. Fatigue tests were performed following a previously validated procedure. The slope part of the Wohler diagram was obtained and a rough endurance limit was estimated for all three formulations. The experimental data showed that the addition of barium sulphate to PMMA bone cement affected the fatigue strength of the material, whereas addition of gentamicin sulphate to the radiopaque PMMA had no effect on the fatigue properties of the bone cement. While PMMA with barium sulphate added was confirmed to have a reduced fatigue strength when compared with plain PMMA, no detrimental effect was found for the addition of gentamicin sulphate to radiopaque PMMA. PMID- 12578215 TI - In vitro strain-induced endothelial cell dysfunction determined by DNA synthesis. AB - Rapid re-endothelialization following balloon angioplasty can reduce restenosis by inhibiting smooth muscle cell migration and proliferation. However, formation of a neointima following angioplasty can be inhibited due to endothelial cell dysfunction and denudation. The purpose of this study was to evaluate mechanical tensile stress as a cause of endothelial cell dysfunction. The Flexercell strain unit was utilized to generate both short-term cyclic and static tensile strain on cultured bovine aortic endothelial cells (BAECs). Before analysis of this loading on BAECs, strain behaviour of the Flexercell system and DNA assay conditions were optimized. This paper demonstrates that, when compared with unloaded controls, 4 h cyclic loading at 4 per cent elongation and 0.1 Hz, and static loading at 4 per cent elongation cause a 44 and 70 per cent decrease in DNA synthesis respectively. In a companion paper, it is demonstrated that low DNA synthesis levels in mechanically loaded cells can be increased by incubation with Ap4A and/or NO donors. PMID- 12578216 TI - P1,P4-diadenosine 5'-tetraphosphate induced DNA synthesis in mechanically injured cultured endothelial cells. AB - Rapid re-endothelialization following balloon angioplasty can reduce restenosis by inhibiting smooth muscle cell migration and proliferation. However, formation of a neointima layer following angioplasty can be inhibited due to endothelial cell dysfunction and denudation. In a companion paper, it has been illustrated that mechanical loading causes a decrease in DNA synthesis in bovine aortic endothelial cells (BAECs) thus rendering them dysfunctional. The purpose of this study was to overcome BAEC dysfunction by incubation with pharmacological agents to increase DNA synthesis. Previous studies demonstrated that the adenosine dinucleotides Ap4A and Ap2A induced nitric oxide (NO) production from BAEC while Ap3A, Ap5A and Ap6A did not. This paper demonstrates that Ap4A and Ap2A induce a 1.46- and 1.16-fold increase in DNA synthesis in mechanically stressed BAECs respectively, while Ap3A, Ap5A and Ap6A do not. Additionally, NOC-18, a slow NO release NO donor, significantly increases DNA synthesis in mechanically stressed BAECs without affecting unloaded cells. These results are consistent with NO inducing DNA synthesis in mechanically stressed BAECs. PMID- 12578217 TI - Cyclic flow characteristics in an idealized asymmetric abdominal aortic aneurysm model. AB - The flow characteristics and the corresponding hydrodynamic stability in an idealized asymmetric abdominal aortic aneurysm (AAA) model have been investigated using a laser Doppler anemometer. A rectified sine waveform was used to simulate aortic flow conditions (Re(delta) = 806 and alpha = 12.2). The flow around the distal neck of the AAA model undergoes transition and becomes turbulent for a fraction of time shortly after the commencement of the deceleration phases at every flow cycle while the rest of the flow inside the model stayed laminar throughout the cycle. As a result of non-symmetric vortical structure development inside the model, the distribution of turbulent shear stresses was found to be highly uneven along the radial direction of the model; this is in contrast to results found by the present authors in the symmetrical AAA model. The maximum turbulent shear stress found at the straight side of the distal neck are four times more than the maximum turbulent shear stress measured at the most dilated side of the distal neck. One of the interesting biological implications of the results is that the outward dilation of the arterial wall may be a physiological response to avoid the high turbulent shear load from the momentarily turbulent blood flow. PMID- 12578218 TI - A proximal femoral implant preserves physiological bone deformation: a biomechanical investigation in cadaveric bones. AB - The aim of this study was to compare the perturbances in bone deformation patterns of the proximal femur due to a conventional cemented femoral stem and a novel uncemented implant designed on the principles of osseointegration. Five matched pairs of fresh frozen human femora were mechanically tested. Bone deformation patterns, measured with a video digitizing system under 1.5 kN joint force, showed that the cemented Spectron femoral implant caused significant alterations to the proximal femoral deformation pattern, whereas the Gothenburg osseointegrated titanium femoral implant did not significantly alter the bone behaviour (p < 0.05). Vertical micromotions measured under 1 kN after 1000 cycles were within the threshold of movement tolerable for bone ingrowth (21 microm for the Gothenburg system and 26 microm for the cemented implant). PMID- 12578219 TI - Contact stresses in the glenoid component in total shoulder arthroplasty. AB - Several studies of retrieved glenoid components from total shoulder arthroplasty show an erosion of the rim, surface irregularities, component fracture and wear resulting from polyethylene deformation in vivo. Particles resulting from polyethylene wear might be one of the reasons for the very high rate of glenoid component loosening found clinically. Because wear can be the result of high contact stresses, the aim of this study is to find out whether or not contact stresses are high enough to cause wear of the glenoid component and what influence the component type and geometry have on polyethylene contact stresses for different humerus abduction angles. Elasticity theory is used in a parametric study of contact stresses in several glenoid component designs. A finite element method is used to confirm the accuracy of the analytical solution. The analysis shows that the peak stress generated in glenoid components under conditions of normal living can be as high as 25 MPa; since this exceeds the polyethylene yield strength, wear and also cold flow of the components can be expected. It is predicted that more conforming components have lower contact stresses, which might result in lower wear rate and less cold flow. It is also found that a metal backed component promotes higher contact stresses than an all-polyethylene component with the same total thickness, therefore it can be expected that metal backed components have inferior wear properties. PMID- 12578220 TI - Metal vs bioabsorbable interference screws: initial fixation. AB - Interference screws are used for graft fixation in anterior cruciate ligament reconstruction. There is current debate as to whether metal or bioabsorbable varieties provide the optimal results. This paper provides a critique of the single load-to-failure uniaxial tensile test data available in the literature, which provide the initial postoperative characteristics. Both metal and bioabsorbable screws were found to give similar levels of pull-out force and stiffness when used on similar types of graft material. Screws with bone-patellar tendon-bone grafts provided the greatest values, exceeding the forces transmitted through the native anterior cruciate ligament during normal daily activities. Hamstring grafts provided lower values, only being comparable with the forces of daily living. Consequently, this raises the question as to whether the rehabilitation regime of the hamstring patient must not be aggressive in the early postoperative stages when using either metal or bioabsorbable screws. This would ensure that the graft will not fail in the initial postoperative period before biological fixation occurs, which will then boost the strength to an acceptable level. PMID- 12578221 TI - Oligodendroglioma: toward molecular definitions in diagnostic neuro-oncology. AB - Oligodendroglial tumors have attracted great interest in both basic and clinical neuro-oncology over the past decade. This interest is mainly due to the clinical observation that anaplastic oligodendrogliomas and anaplastic oligoastrocytomas, in contrast to the vast majority of anaplastic astrocytomas and glioblastomas, frequently respond favorably to chemotherapy. In addition, oligodendroglial tumors are associated with longer survival times than the diffuse astrocytic gliomas. These differences in response to therapy and in prognosis have been associated with distinct genetic aberrations, in particular the frequent loss of alleles on chromosome arms 1p and 19q in oligodendroglial tumors. In addition, other genetic changes have been reported as indicators of poor response to therapy and short survival, including homozygous deletion of the CDKN2A gene at 9p21, mutation of the PTEN gene at 10q23, and amplification of the EGFR gene at 7p12. In this review we summarize the current state of the art concerning the molecular genetics of oligodendroglial tumors. A particular focus is placed on the role of molecular genetic findings in the diagnostic and prognostic assessment of these neoplasms. As a result of the recent advances in the field, we propose that clinical decisions in the management of patients with oligodendroglial tumors should be based on the combined assessment of clinical and neuroimaging features, histological classification and grading, as well as molecular genetic characteristics. PMID- 12578222 TI - Inflammatory mechanisms after ischemia and stroke. AB - Inflammation has been implicated as a secondary injury mechanism following ischemia and stroke. A variety of experimental models, including thromboembolic stroke, focal and global ischemia, have been used to evaluate the importance of inflammation. The vasculature endothelium promotes inflammation through the upregulation of adhesion molecules such as ICAM, E-selectin, and P-selectin that bind to circulating leukocytes and facilitate their migration into the CNS. Once in the CNS, the production of cytotoxic molecules may facilitate cell death. The macrophage and microglial response to injury may either be beneficial by scavenging necrotic debris or detrimental by facilitating cell death in neurons that would otherwise recover. While many studies have tested these hypotheses, the importance of inflammation in these models is inconclusive. This review summarizes data regarding the role of the vasculature, leukocytes, blood-brain barrier, macrophages, and microglia after experimental and clinical stroke. PMID- 12578223 TI - In vivo multiphoton imaging of a transgenic mouse model of Alzheimer disease reveals marked thioflavine-S-associated alterations in neurite trajectories. AB - Postmortem analyses of senile plaques reveal numerous dystrophic processes in their vicinity. We used in vivo multiphoton microscopy of a transgenic model of Alzheimer disease (AD) to simultaneously image senile plaques and nearby neuronal processes. Plaques were labeled by immunofluorescent staining or thioflavine-S and neuronal processes were labeled with a fluorescent dextran conjugate. Imaging of 3-dimensional volumes in the vicinity of plaques revealed subtle changes in neurite geometry in or near diffuse plaques. By contrast, disruptions in neurite morphology, including dystrophic neurites immediately surrounding plaques as well as major alterations in neurite trajectories, were seen in association with thioflavine-S-positive plaques. Nearly half of all labeled processes that came within 50 microm of a thioflavine-S-positive plaque were altered, suggesting a fairly large "halo" of neuropil alterations that extend beyond the discrete border of a thioflavine-S plaque. These results support the hypothesis that compact thioflavine-S-positive plaques disrupt the neuropil in AD. PMID- 12578225 TI - The microvasculature of the cerebral white matter: arteries of the subcortical white matter. AB - The microvascular architecture of the human cerebral subcortical white matter was studied. Most of the subcortical arteries ran straight through the cortex, but upon entering the white matter, they began to coil, loop, and spiral. Vascular stains showed wide spaces between the adventitial sheaths and blood vessels. The blood vessels coiled, looped, and spiraled within these wide adventitial spaces. This phenomenon was observed in the brains from persons ranging from the first to ninth decades of life and there were no statistically significant age-related correlations. Furthermore, there was no evidence of a reduction in the volume of white matter after fixation. Therefore, the observed tortuosity does not appear to be the result of shrinkage of brain tissue following fixation. While the mechanisms responsible for the subcortical arteries circuitry remain undetermined, this coiling architecture may serve as a trap for tumor cells and microorganisms passing through the blood stream, suggesting that these coiling arterial blood vessels may play a significant role in the pathogenesis of tumor metastasis and the brain abscess that frequently occurs in the gray-white matter junction. PMID- 12578224 TI - Mercapturate metabolism of 4-hydroxy-2-nonenal in rat and human cerebrum. AB - 4-Hydroxy-2-nonenal (HNE), a potent toxin formed in the brain from oxidation of polyunsaturated fatty acids, is increased in Alzheimer disease (AD), where it is a proposed effector of amyloid beta peptide-mediated neurotoxicity. Detoxification of HNE via the mercapturic acid pathway (MAP) is the primary means by which other organs, such as liver, limit its toxic effects. Here we examined the distribution and activity of MAP detoxification for HNE in cerebrum. Our results showed that rat cerebral cortex and especially synaptosomes were less well equipped to detoxify HNE via the MAP than liver. Glutathione transferases (GSTs) catalyze the committed step in the MAP; GST-mu and GST-pi, but not OST alpha, were detected in neurons and astrocytes in cerebrum from AD patients and controls. MAP activity in frontal cortex of AD patients was modestly but significantly increased compared to controls. These data suggest that lipid peroxidation may present a greater toxic burden to cerebrum than to other organs, and that a component of response to injury in late stage AD is a slight increase in MAP activity. PMID- 12578226 TI - Oxidized galectin-1 stimulates the migration of Schwann cells from both proximal and distal stumps of transected nerves and promotes axonal regeneration after peripheral nerve injury. AB - Oxidized galectin-1 has recently been identified as a key factor that plays important roles in initial axonal growth in injured peripheral nerves. The aim of this study was to investigate the effects of oxidized galectin-1 on regeneration of rat spinal nerves using acellular autografts (containing no viable cells) and allografts (containing no cell membranes) with special attention to the relationship between axonal regeneration and Schwann cell migration. Immunohistochemically, endogenous galectin-1 was expressed in dorsal root ganglion (DRG) neurons, spinal cord motoneurons, and axons and Schwann cells in normal sciatic nerves. Administration of oxidized recombinant human galectin-1 (rh-gal-lox, 5 ng/ml) in autograft model promoted axonal regeneration from motoneurons as well as from DRG neurons; this was confirmed by a fluorogold tracer study (p < 0.05). Anti-rh-gal-1 antibody (30 microg/ml) strongly inhibited axonal regrowth (p < 0.05). Pretreatment of allografts with rh-gal-lox stimulated the migration of Schwann cells not only from proximal stumps but also from distal stumps into the grafts, resulting in accelerated axonal regeneration (p < 0.05). Moreover, Schwann cell migration preceded the axonal growth in the presence of exogenous rh-gal-lox in the grafts. These results strongly suggest that local administration of exogenous rh-gal-lox promotes the migration of Schwann cells followed by axonal regeneration from both motor and sensory neurons, resulting in acceleration of neuronal repair. This technique may also be of value in the repair of human nerves. PMID- 12578227 TI - Transglutaminase activity, protein, and mRNA expression are increased in progressive supranuclear palsy. AB - Transglutaminases catalyze the covalent cross-linking of substrate proteins to form insoluble protein complexes that are resistant to degradation. Our previous studies demonstrated that transglutaminase-induced cross-linking of tau proteins occurs in Alzheimer disease and progressive supranuclear palsy (PSP). The current study was designed to measure transglutaminase enzyme activity and the mRNA and protein levels of 3 transglutaminase isoforms that are expressed in human brain. Overall, transglutaminase activity was significantly increased in the globus pallidus (182% of control) and pons in PSP (171% of control) but not the occipital cortex (a region spared from pathology). Using a Spearman rank correlation test, we found that tissues with more transglutaminase-activity had more neurofibrillary tangles. Protein and mRNA levels of transglutaminase 1 were increased in globus pallidus of PSP as compared to controls. There were also significantly higher mRNA levels of the short form of transglutaminase 2 in globus pallidus of PSP (974% of control). Transglutaminase 1 mRNA and the long isoform of transglutaminase 2 mRNA (2212% of control) were significantly higher in PSP in the dentate of cerebellum. Together, these findings suggest that transglutaminase 1 and 2 enzymes may be involved in the formation and/or stabilization of neurofibrillary tangles in selectively vulnerable brain regions in PSP. These transglutaminases may be potential targets for therapeutic intervention. PMID- 12578228 TI - Ependymal cell reactions in spinal cord segments after compression injury in adult rat. AB - Recently, it has been suggested that neural stem cells and neural progenitor cells exist in the ependyma that forms the central canal of the spinal cord. In this study, we produced various degrees of thoracic cord injury in adult rats using an NYU-weight-drop device, assessed the degree of recovery of lower limb motor function based on a locomotor rating scale, and analyzed the kinetics of ependymal cell proliferation and differentiation by proliferating cell nuclear antigen (PCNA), nestin, glial fibrillary acidic protein (GFAP), or GAP-43 immunostaining. The results showed that the time course of the ependymal cell proliferation and differentiation reactions differed according to the severity of injury, and that the responses occurred not only in the neighborhood of the injury but in the entire spinal cord. An increase in the locomotor rating score was related to an increase in the number of PCNA-positive cells, and the differentiation of ependymal cells into reactive astrocytes was involved in injury repair. No apoptotic cells in the ependyma were detectable by the TUNEL method. These results indicate that the ependymal cells of the spinal central canal are themselves multipotent, can divide and proliferate according to the severity of injury, and differentiate into reactive astrocytes within the ependyma without undergoing apoptosis or cell death. PMID- 12578229 TI - Reversal of thiamine deficiency-induced neurodegeneration. AB - Neurodegenerative diseases are characterized by abnormalities in oxidative processes, region-selective neuron loss, and diminished thiamine-dependent enzymes. Thiamine deficiency (TD) diminishes thiamine dependent enzymes, alters mitochondrial function, impairs oxidative metabolism, and causes selective neuronal death. In mice, the time course of TD-induced changes in neurons and microglia were determined in the brain region most sensitive to TD. Significant neuron loss (29%) occurred after 8 or 9 days of TD (TD8-9) and increased to 90% neuron loss by TD10-11. The number of microglia increased 16% by TD8 and by nearly 400% on TD11. Hemeoxygenase-1 (HO-1)-positive microglia were not detectable at TD8, yet increased dramatically coincident with neuron loss. To test the duration of TD critical for irrevocable changes, mice received thiamine after various durations of TD. Thiamine administration on TD8 blocked further neuronal loss and induction of HO-1-positive microglia, whereas other microglial changes persisted. Thiamine only partially reversed effects on TD9, and was ineffective on TD10-11. These studies indicate that irreversible steps leading to neuronal death and induction of HO-1-positive microglia occur on TD9. The results indicate that TD induces alterations in neurons. endothelial cells, and microglia contemporaneously. This model provides a unique paradigm for elucidating the molecular mechanisms involved in neuronal commitment to neuronal death cascades and contributory microglial activity. PMID- 12578230 TI - Apoptotic PC12 cells exposing phosphatidylserine promote the production of anti inflammatory and neuroprotective molecules by microglial cells. AB - The interaction of phosphatidylserine (PS), exposed on the surface of apoptotic cells and with its specific receptor (PtdSerR) expressed by microglia, is a crucial event in the recognition and clearance of apoptotic neurons. Here, we extend our previous studies in which PS-liposomes mimicking apoptotic cells were used to investigate the functional role of PS-PtdSerR interactions on microglial functional state. Purified rat microglial cells were either incubated with PC12 cells maintained in complete medium (healthy), exposed to staurosporine or serum deprivation (apoptotic), or treated with hydrogen peroxide (necrotic). After 24 hours, supernatants from co-cultures and single cell type cultures were analyzed for nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), prostaglandin E2 (PGE2), transforming growth factor-beta1 (TGF-beta1), and nerve growth factor (NGF). When lipopolysaccharide (LPS)-activated microglia was cultured with apoptotic PC12 cells, NO and TNF-alpha levels significantly decreased, IL-10 was not affected, and PGE2 levels were substantially increased. In addition, TGF-beta and NGF syntheses increased when resting microglia was cultured with apoptotic but not healthy or necrotic PC12 cells. We proposed that upon interaction with PS-expressing apoptotic neurons, microglia no longer act as a promoter of the inflammatory cascade and that the specific microglial functional state induced by PS-PtdSerR may be relevant for the final outcome of neurodegenerative diseases. PMID- 12578231 TI - Not knowing. PMID- 12578232 TI - Early neurodevelopmental markers predictive of mortality in infants infected with HIV-1. AB - One-hundred and fifty-seven vertically infected HIV-1 positive infants (85 males, 72 females) underwent longitudinal assessment to determine whether early neurodevelopmental markers are useful predictors of mortality in those infants who survive to at least 4 months of age. Survival analysis methods were used to estimate time to death for quartiles of 4-month scores (baseline) on the Bayley Scales of Infant Development (BSID). Cox proportional hazards progression was used to estimate relative hazard (RH, 95% CI) of death for BSID scores and potential confounders. Thirty infants with BSID scores at 4 months of age died during follow-up. Survival analysis revealed greater mortality rates in infants with BSID (Mental Developmental Index and Psychomotor Developmental Index) scores in the lower quartile (p=0.004, p=0.036). Unadjusted univariate analyses revealed increased mortality associated with baseline CD4+ 29%, gestational age <37 weeks, smaller head circumference, advanced HIV and higher plasma viral load. BSID scores independently predicted mortality after adjusting for treatment, clinical category, gestational age, plasma viral load and CD4+ percentage. PMID- 12578233 TI - Hand function in children with hemiplegic cerebral palsy: prospective follow-up and functional outcome in adolescence. AB - The aim of this prospective study was to determine the outcome of affected hand impairment and disability in a group of 31 children (16 males, 15 females) with hemiplegic cerebral palsy who were referred consecutively to our rehabilitation service and followed from a mean age of 2 years and 7 months to a mean age of 12 years and 9 months. The hand function assessment protocol consisted of video recorded procedures: one to assess grip and the other to assess the extent of spontaneous use of the affected hand. Assessments at outset and at latest follow up showed that the series as a whole had greater hand impairment in spontaneous manipulation than in gripping tasks. Comparison of hand function before age 4 years with the latest assessment over age 11 years revealed a non-significant improvement in grip, whereas spontaneous hand use remained stable. All but one of the children who had good grip and hand use scores at first assessment maintained them over time unless they were not treated for hand dysfunction. Among the children with low scores at outset, the improvement over time was more marked in grip than spontaneous hand use and occurred mainly in the early years. These results suggest that to evaluate the real disability of the affected hand in children with hemiplegia, grip assessment is insufficient and that an instrument assessing spontaneous hand use in bilateral manipulation is required. Furthermore, intensive treatment focused on hand function should be planned in the early years for children with more severe hand impairment, whereas regular follow-up is sufficient for less affected children. PMID- 12578234 TI - Melbourne Assessment of Unilateral Upper Limb Function: construct validity and correlation with the Pediatric Evaluation of Disability Inventory. AB - The Melbourne Assessment of Unilateral Upper Limb Function (Melbourne Assessment) is an evaluation tool that objectively measures upper-extremity function in children with cerebral palsy (CP). This study investigates how well performance on the Melbourne Assessment relates to the child's actual performance in functional tasks. Eighteen children with CP (5 to 14 years of age; nine males, nine females) were assessed using the Melbourne Assessment and the Pediatric Evaluation of Disability Inventory (PEDI). Five children had spastic quadriplegia, eight had spastic diplegia, two had spastic hemiplegia and diplegia, two had athetosis, and one had hypotonic quadriplegia with mobile ventilator dependence. Children's performances were statistically correlated using Spearman's rho to establish the relation between these tools. Very high correlation coefficients were calculated between the Melbourne Assessment and self-care (0.939) and mobility domains (0.783) of the PEDI and the overall functional skills section of the PEDI (0.718). The Melbourne Assessment demonstrates excellent construct validity for upper limb functioning. PMID- 12578235 TI - Motor, cognitive, and behavioural disorders in children born very preterm. AB - Children born preterm have been shown to exhibit poor motor function and behaviour that is associated with school failure in the presence of average intelligence. A geographically determined cohort of two-hundred and eighty preterm children (151 males, 129 females) born before 32 weeks' gestation and attending mainstream schools were examined at 7 to 8 years of age together with 210 (112 males, 98 females) age- and sex-matched control participants were tested for motor, cognitive, and behavioural problems. Tests applied were the Movement Assessment Battery for Children (MABC), Clinical Observations of Motor and Postural Skills (COMPS), Developmental Test of Visual-Motor Integration (VMI), Wechsler Intelligence Scale for Children, and Connors' Teacher Rating Scale for attention-deficit-hyperactivity disorder (ADHD). Control children scored significantly better than the preterm group on all motor, cognitive, and behavioural measures. The lowest birthweight and most preterm individuals tended to score the lowest. Motor impairment was diagnosed in 86 (30.7%) of the preterm group and 14 (6.7%) of the control children using the MABC; 97 (42.7%) and 18 (10.2%) using the COMPS; and 68 (24.3%) and 17 (8.1%) respectively using the VMI. Each test of motor function identified different children with disability, although 23 preterm children were identified as having motor disability by all three tests. Preterm children were more likely to have signs of inattention and impulsivity and have a diagnosis of ADHD. Minor motor disabilities persist in survivors of preterm birth despite improvements in care and are not confined to the smallest or most preterm infants. They may exist independently of cognitive and behavioural deficits, although they often co-exist. The condition is heterogeneous and may require more than one test to identify all children with potential learning problems. PMID- 12578236 TI - Perinatal morbidity, mild motor delay, and later school outcomes. AB - Four dimensions of motor competence were evaluated in 4-year-old term and preterm children and were related to academic achievement and use of school services at age 8 years. The objective of the study was to evaluate a graduated 'stair-step' effect between perinatal morbidity, mild motor delay, and later school outcomes in 168 children (88 females, 80 males) stratified into four study groups: 134 healthy term infants > or = 37 weeks' gestation, birthweight > or = 2500g); 134 preterm infants < or = 37 weeks' gestation divided into healthy preterm (n=41), clinically ill preterms (n=59), and preterm infants with neurological illness (n=34). Significant differences were found in total, fine and gross motor performance, and visual-motor integration at age 4 years. Scores for the preterm groups decreased with increasing morbidity. At age 4 years, mild motor delay was found in all preterm groups. Children with mild motor delay had lower academic achievement scores (Wide Range Achievement Test-3) and higher rates of school service use at age 8 years. Perinatal morbidity and compromised motor performance are important precursors of educational underachievement. PMID- 12578238 TI - Opioid peptides and dipeptidyl peptidase in autism. AB - It has been hypothesized that autism results from an 'opioid peptide excess'. The aims of this study were to (1) confirm the presence of opioid peptides in the urine of children with autism and (2) determine whether dipeptidyl peptidase IV (DPPIV/CD26) is defective in children with autism. Opioid peptides were not detected in either the urine of children with autism (10 children; nine males, one female; age range 2 years 6 months to 10 years 1 month) or their siblings (10 children; seven males, three females; age range 2 years 3 months to 12 years 7 months) using liquid chromatography-ultraviolet-mass spectrometric analysis (LC UV-MS). Plasma from 11 normally developing adults (25 years 5 months to 55 years 5 months) was also tested. The amount and activity of DPPIV in the plasma were quantified by an ELISA and DPPIV enzyme assay respectively; DPPIV was not found to be defective. The percentage of mononuclear cells expressing DPPIV (as CD26) was determined by flow cytometry. Children with autism had a significantly lower percentage of cells expressing CD3 and CD26, suggesting that they had lower T cell numbers than their siblings. In conclusion, this study failed to replicate the findings of others and questions the validity of the opioid peptide excess theory for the cause of autism. PMID- 12578237 TI - Effect of environmental setting on mobility methods of children with cerebral palsy. AB - The aim of this study was to: (1) describe the usual mobility methods of children with cerebral palsy (CP) at home, school, and outdoors or in the community and (2) examine whether children with CP are more dependent on adult assistance for mobility in certain settings. The participants were a stratified random sample of 636 children with CP (355 males and 281 females; 2 to 12 years of age, mean 6.8 years SD 2.7), receiving rehabilitation services in Ontario, Canada. Children were grouped by age and Gross Motor Function Classification System (GMFCS) level. Among the five levels of the GMFCS, there were 185 children classified at level I, 81 children at level II, 113 children at level III, 132 children at level IV, and 125 children at level V. Information on children's usual mobility was obtained by parent report. The results of logistic regression indicated that compared with the school setting, children were more dependent on adult assistance for mobility when outdoors/in the community and less dependent at home. The majority of children aged from 4 to 12 years at levels III to V used wheelchair mobility at school and outdoors or in the community, however, only a small percentage self-propelled their wheelchair or used powered mobility. Of the children aged 4 to 12 years at level V, 39% were carried at home. The findings suggest that environmental setting is an important consideration for assessment and intervention to improve mobility of children with CP. For children who do not walk, attention should be given to the needs of caregivers and factors that are important for successful powered mobility. PMID- 12578239 TI - Community integration among adults with spinal cord injuries sustained as children or adolescents. AB - The purpose of this study was to determine factors associated with community integration of adults who sustained spinal cord injuries as children or adolescents. Outcome measures included the Craig Handicap Assessment and Reporting Technique and measures of independent living. Independent variables include demographic factors, level of injury, functional independence as measured by the Functional Independence Measure, and perceived health as measured by Short Form-12 physical and mental component scores. Data were analyzed by multiple regression analyses for each of the outcome measures. Two-hundred and sixteen adults (150 males and 66 females) were interviewed. Mean age at interview was 29 years (range 24 to 27 years) and mean age at injury was 14 years (range 0 to 18 years). Results showed that the five factors most strongly associated with community integration were education level, employment, income, functional independence, and health status. Factors that were not associated with community integration included sex, age at injury, duration, and level of injury. PMID- 12578240 TI - The bases of western attitudes to consanguineous marriage. PMID- 12578241 TI - Facial naevus flammeus with choroidal haemangioma and without intracranial involvement. PMID- 12578242 TI - Salivary cortisol in children with cognitive impairment. PMID- 12578243 TI - Effect of practice on hand preference on a color visual discrimination task by capuchin monkeys (Cebus apella). AB - Effects of practice on hand preference in nonhuman primates have indicated an initial right-hemispheric advantage in new tasks that can be reversed or disappear after a long period of training. The aim of this study was to examine the effects of 6-mo. practice on hand preference in capuchin monkeys (Cebus apella) performing a color visual discrimination task which requires attention and learning or memory processes. For this purpose, an early period of practice (first 200 trials) was compared with a late period of practice (the last 200 trials after 6 mo. practice). No population-level biases were observed between the use of hands either early in training or after 6 mo. of practice. Individual analysis indicated that two of the five subjects tested strengthened their initial hand preference but three subjects changed their preference with the practice. No correlation was observed between hand use and performance. These results suggest that hand preference in capuchin monkeys can be influenced by practice and is not only a direct function of hemispheric specialization. PMID- 12578244 TI - Ventilatory thresholds in arm and leg exercises with spontaneously chosen crank and pedal rates. AB - The present study assessed whether the first and the second ventilatory thresholds (VT1 and VT2) were dependent on the muscle groups solicited when spontaneously chosen crank and pedal rates are used. 20 physical education male students (22 +/- 2.2 yr.) performed two maximal incremental tests randomly assigned using an increment of 15 and 30 W every minute for arm and leg exercises, respectively. These tests were used to measure the maximal oxygen uptake (VO2 max) and to identify VT1 and VT2. The absolute oxygen uptake (VO2) values measured at VT1, VT2, and at maximal workload were significantly (p < .05) lower during arm and leg exercises. However, VT1 and VT2 expressed in percent of VO2 max were not significantly different between arm and leg exercises (54.1 +/- 8.2 vs 57.2 +/- 11.4%; and 82.5 +/- 6.4 vs 84.6 +/- 5.1% at VT1 and VT2, respectively). In addition, at the two thresholds, none of the variables measured during arm and leg exercises were significantly correlated with the exception of spontaneously chosen crank and pedal rates (p < .01; r = .75 and r = .69 for VT1 and VT2, respectively). Probably due to the different training status and skill level, no extrapolation can be made to specify the arm thresholds from the leg. These results underline the need to specify the ventilatory thresholds from specific arm ergometer measures obtained from tests performed with spontaneously chosen crank and pedal rates and, thus, close to sport and recreational activities, when they are used for training and rehabilitation programs. PMID- 12578245 TI - An assessment of the validity and reliability of two perceived exertion rating scales among Hong Kong children. AB - This study evaluated the validity and reliability of the Chinese-translated (Cantonese) versions of the Borg 6-20 Rating of Perceived Exertion (RPE) scale and the Children's Effort Rating Table (CERT) during continuous incremental cycle ergometry with 10- to 11-yr.-old Hong Kong school children. A total of 69 children were randomly assigned, with the restriction of groups being approximately equal, to two groups using the two scales, CERT (n = 35) and RPE (n = 34). Both groups performed two trials of identical incremental continuous cycling exercise (Trials 1 and 2) 1 wk, apart for the reliability test. Objective measures of exercise intensity (heart rate, absolute power output, and relative oxygen consumption) and the two subjective measures of effort were obtained during the exercise. For both groups, significant Pearson correlations were found for perceived effort ratings correlated with heart rate (rs > or = .69), power output (rs > or = .75), and oxygen consumption (rs > or = .69). In addition, correlations for CERT were consistently higher than those for RPE. High test retest intra-class correlations were found for both the effort (R = .96) and perceived exertion (R = .89) groups, indicating that the scales were reliable. In conclusion, the CERT and RPE scales, when translated into Cantonese, are valid and reliable measures of exercise intensity during controlled exercise by children. The Effort rating may be better than the Perceived Exertion scale as a measure of perceived exertion that can be more validly and reliably used with Hong Kong children. PMID- 12578246 TI - Susceptibility to illusions and cognitive style: implications for pharmacy dispensing. AB - Response distributions for five cognitive illusions and one visual illusion were examined in two samples, college students (n = 134) and pharmacists (n = 51). These illusions were selected for study on the basis of pharmacists' judgments about associations of illusions to common dispensing errors. Participants were categorized as Illusion-prone or Illusion-resistant, and distributions of such tendencies for the six stimuli used varied within samples. Significant differences between the two samples on illusion-proneness and resistance were observed for the "Moses' Ark" and -Fcount" illusions. Associations of Illusion prone and Illusion-resistant responses to field-dependence, psychological type, and the cognitive orientations derived from Psychological Type Theory were examined. Field-independence-field-dependence was the only cognitive dimension associated with Illusion-prone and Illusion-resistant responding. Implications of the data for developing measures based upon visual and cognitive illusions to identify people with error-prone tendencies were discussed. PMID- 12578247 TI - Selective, sustained, and shift in attention in patients with diagnoses of schizophrenia. AB - Attentional deficits are a prominent aspect of cognitive dysfunction in schizophrenia. The present study was designed to investigate attention deficit in a group of patients with diagnosis of schizophrenia. According to the segmental set theory suggested by Hogarty and Flesher, three aspects of attention problems, selective, sustained, and shift in attention, were studied. The 30 patients hospitalized on three psychiatric wards at Shiraz and Isfahan and 30 normal healthy subjects matched for age, sex, and years of education were administered a computerized Continuous Performance Test, Stroop Color-word Test, and Wisconsin Card Sorting test. Analysis showed patients performed more poorly than control subjects on measured aspects of attention. The acute/chronic classification did not predict differences in attention scores between subtypes of schizophrenia, while the positive/negative classification did. Paranoid, undifferentiated, and residual groups by subtypes of schizophrenia showed similar performance on the Continuous Performance Test, but were significantly different on errors on the Wisconsin Card Sorting test and on reaction time to Stroop stimuli in the incongruent color-word condition. Patients with paranoid diagnosis performed better than other subtypes on these tasks. Present results suggest that the Continuous Performance Test is valuable for differentiating of schizophrenia spectrum disorder, while scores on Stroop and Wisconsin card sorting may have better diagnostic value for differentiating subtypes of the disorder. PMID- 12578248 TI - Note on Kazlauskaite and Lynn's paper on retest reliability of the Colored Progressive Matrices. AB - A reanalysis of the retest reliabilities for the Colored Progressive Matrices indicates Kazlauskaite and Lynn's conclusions (2002) were not accurate. PMID- 12578249 TI - Effect of metaphorical verbal instruction on modeling of sequential dance skills by young children. AB - Metaphorical verbal instruction was compared to specific verbal instruction about movement in the modeling of sequential dance skills by young children. Two groups of participants (Younger, mean age 5:3 yr., n = 30: Older, mean age 6:2 yr., n = 30) were randomly assigned to conditions in a 2 (sex) x 2 (age [Younger and Older]) x 3 (verbal instruction [Metaphorical, Movement-relevant, and None]) factorial design. Order scores were calculated for both performance and recognition tests, comprising five acquisition trials and two retention trials after 24 hr., respectively. Analysis of variance indicated that the group given metaphorical instruction performed better than the other two instructions for both younger and older children. The results suggest that metaphorical verbal instruction aids the recognition and performance of sequential dance skills in young children. PMID- 12578250 TI - Increased visual dependence in Parkinson's disease. AB - The present study tested the hypothesis that there is increased visual dependence perceptually in patients with Parkinson's disease. We also evaluated whether the visual control of posture and locomotion was related to perceptual visual field dependence. 21 patients with idiopathic Parkinson's disease and 22 age-matched normal subjects were compared on judgment of the visual vertical using the Rod and-Frame test with visual perturbations in the frontal plane with a tilted frame. Patients had significantly larger errors than controls in the estimation of the subjective vertical. In the same experiment, we performed a posture and a gait analysis in both groups. Posturographic evaluation did not indicate significant differences in unsteadiness between patients and controls. Gait analysis indicated a typical pattern of reduced velocity, shortened stride length, and normal step width. A significant correlation of .89 was found only in the Parkinsonian group between their errors in estimating subjective visual vertical and the Romberg quotient evaluating visual contribution to postural control. No specific locomotor pattern was correlated with visual dependence. Considering our results and previous reports on the visual control of posture, we conclude that patients with Parkinson's disease showed a significantly increased dependence upon visual information both perceptually and motorically, with an increased perceptual visual dependence in the patients being predictive of an equivalent visual dependence or visual control of posture and equilibrium. PMID- 12578251 TI - Visual degradation in Boston Naming Test performance. AB - The 60 pictures of the Boston Naming Test were degraded via a mask to be either "easy," "medium", or "hard" to process visually. The visual mask was created by cutting a 2-in. x 2-in. square from a security mailing envelope and progressively reducing the size 25% on a copy machine. Three groups of 50 undergraduates each were randomly assigned to one of these three degradation conditions, and the number of correct identifications made was measured. We expected that as degradation increased (from easy to medium to hard), Boston Naming Test performance would decrease. A one-way analysis of variance indicated that group scores differed. With "easy" degradation more pictures were identified in comparison with performance in the "hard" degradation condition, but no other differences were significant. Results were discussed from the perspective of how unknown situational variables may affect performance on psychological tests. PMID- 12578252 TI - Idiosyncratic reality claims, relaxation dispositions, and ABC relaxation theory: happiness, literal christianity, miraculous powers, metaphysics, and the paranormal. AB - This study examined idiosyncratic reality claims, that is, irrational or paranormal beliefs often claimed to enhance relaxation and happiness and reduce stress. The Smith Idiosyncratic Reality Claims Inventory and the Smith Relaxation Dispositions Inventory (which measures relaxation and stress dispositions, or enduring states of mind frequently associated with relaxation or stress) were given to 310 junior college student volunteers. Principal components factor analysis with varimax rotation identified five idiosyncratic reality claim factors: belief in Literal Christianity; Magic; Space Aliens: After Death experiences; and Miraculous Powers of Meditation, Prayer, and Belief. No factor correlated with increased relaxation dispositions Peace, Energy, or Joy, or reduced dispositional somatic stress, worry, or negative emotion on the Smith Relaxation Dispositions Inventory. It was concluded that idiosyncratic reality claims may not be associated with reported relaxation, happiness, or stress. In contrast, previous research strongly supported self-affirming beliefs with few paranormal assumptions display such an association. PMID- 12578253 TI - Jaw and finger movement accuracy under visual and nonvisual feedback conditions. AB - The present study revisited the issue whether the presence of added visual feedback differentially affects the accuracy of finger and jaw movements. 15 men were instructed to move either the index finger on the dominant (right) hand, or the jaw, to a predefined target position with the highest precision possible. During execution of the task, on-line visual feedback of the moving articulator was either present or removed. In contrast to previous findings, significant improvement was observed for both finger and jaw movements in the visual feedback condition. Movement error in the nonvisual condition was proportionally greater for finger than for jaw movements which may have reflected a speed-accuracy trade off because finger movements in the nonvisual condition were executed significantly faster than those of the jaw. The present findings support the beneficial effects of adding visual feedback during dynamic oral and finger movements that require a high spatial precision. Such findings support current methods of clinical intervention in speech-language pathology anl other disciplines. Furthermore, the results contribute to our understanding of the role of various modalities of feedback during motor execution. PMID- 12578254 TI - Relation between sport activity and mental rotation: comparison of three groups of subjects. AB - The aim was to observe the relation between sport activity and performance on a mental image-transformation task. A classical mental rotation task using abstract stimuli was administered to three groups: (a) gymnasts who used mental and physical rotations in their practice, (b) athletes whose activities required very little motor rotation, and (c) nonathletes. Both sport groups performed similarly and obtained significantly shorter response times than those of the nonathletes. We suggest that the regular practice of spatial activities, such as sports, could be related to the spatial capacities of the participants. PMID- 12578255 TI - Task duration in contextual interference. AB - Duration of practice trial on a pursuit rotor task in contextual interference was investigated. Participants practiced at each of 4 angular velocities, with 24 participants completing 28 trials lasting 20 sec., and 24 participants completing 112 trials of 5 sec. Half of the participants in each trial-duration condition practiced in a blocked format and half practiced in a random format. After random practice posttest performance was better than blocked practice when practice trial duration was 20 sec., but worse when practice-trial duration was 5 sec. This result is not consistent with theoretical explanations of the contextual interference effect and is discussed with reference to the task characteristics and demands of the pursuit rotor. PMID- 12578256 TI - Young children's change in strategies of size comparison: effects of strategy evaluation. AB - This study examined how young children change their strategies for comparing sizes. 33 5- and 6-yr.-old children were asked to compare the sizes of geometric figures, and their strategies were observed. Children in experimental conditions were also asked to evaluate presented strategies for comparing geometric figures, and half of the children were provided with feedback on the evaluation. Analysis showed children provided with feedback on evaluation showed increased use of the strategy of placing one figure on another. PMID- 12578257 TI - Acute changes in selected serum enzyme and metabolite concentrations in 12- to 14 yr.-old athletes after an all-out 100-m swimming sprint. AB - The purpose of this study was to investigate the acute effects of an all-out 100 m swimming sprint on changes in serum enzyme and calcium ion concentrations in young (12 to 14 years) male and female swimmers. Changes in serum enzyme concentrations of creatine kinase (CK), lactate dehydrogenase (LDH), glutamic pyruvate transaminase (GPT), glutamic oxaloacetate transaminase (GOT), serum glucose (GL), and calcium ion ((Ca2+) concentrations were measured in 23 elite swimmers (13 boys and 10 girls) before and after a 100-m freestyle all-out sprint. Analysis showed (1) there were significant sex differences in serum CK concentration at baseline (Pretest); (2) significant sex differences in serum CK and LDH concentrations after the 100-m spring; (3) no significant differences in Ca2+ concentration after the 100-m spring; and (4) significant increase in Serum GOT and blood glucose concentrations after the 100-m sprint, suggesting that these might both be useful indicators of anaerobic exercise stress in young swimmers. PMID- 12578258 TI - Exploratory factor analysis of the Sport Orientation Questionnaire and the Task and Ego Orientation in Sport Questionnaire in a Japanese sport setting. AB - This study examined sport achievement orientation within the japanese sport setting. 1,836 male and 425 female athletes (M age = 18.6 yr.) from 47 sports completed the Japanese version of the Sport Orientation Questionnaire (24 items), and 1,781 males and 421 females (M age = 18.6 yr.) from 47 sports completed that of the Task and Ego Orientation in Sport Questionnaire (14 items). While the original English versions of the questionnaires are composed of three and two factors, respectively, present exploratory factor analyses identified four factors in the Sport Orientation Questionnaire and three in the Task and Ego Orientation in Sport Questionnaire. In this study, two types of the SOQ Competitiveness and the TEOSQ Ego Orientation emerged even though the original versions included only one type. A one-way analysis of variance indicated that sex differences were significant for all subscales. Sex differences in the Sport Orientation Questionnaire of Japanese resembled those of Americans. The overall factor reliability and validity based on the sample suggested that the modified Japanese versions of the two questionnaires can be valuable in the investigation of sport achievement orientation in Japanese sport and exercise settings. PMID- 12578259 TI - Discriminating overhead points of contact after arm raising. AB - Arm-raising movements at the shoulder made to overhead locations were investigated in a weight-hearing stance with forward vision available. When discrimination of different points of contact was measured after arm elevations to the unseen overhead targets, no dominant arm advantage was found, but contacts made after the shortest (90 degrees) arm-elevation movements were better discriminated than those from arm-at-side (180 degrees), plus windup (540 degrees). PMID- 12578260 TI - Visual and lexical factors in naming speed by children with reading disorders. AB - Reaction times of 12 reading disordered and 12 normally reading children and 12 adults were investigated with a visual half-field tachistoscopic picture-naming task. Analysis indicated slower picture-naming times for children with reading disorders. Hemispheric processing models suggested neurolinguistic maturation for rapid picture-naming speed. PMID- 12578261 TI - Effects of appropriate and inappropriate odors on product evaluations. AB - In accounting for an odorant's effect on the evaluation of a product, both the odor's intrinsic pleasantness and its appropriateness for that particular product are relevant. When comparing the effects of pleasant smells, consumers are likely to prefer products with appropriate smells to those with an inappropriate smell. We investigated the effect of adding congruent and incongruent odorants on product evaluations for each of three product categories: food (tea, instant whip, cake mix), personal care (shampoo, deodorant, lip balm), and household products (cream cleaner, air freshener, furniture wax). In a between-subjects design, 96 respondents judged scented and unscented products presented in commercial packages of major national brands. The respondents assessed the overall evaluation, 14 to 19 beliefs about the product, buying intention, and the frequency of use for each product. Respondents were instructed to evaluate each product as they would in a store. Although congruency ratings between odor and product show that the manipulation of congruency was successful, no main effect was found for the congruency on overall evaluations of the products. To account for this unexpected finding, we speculate that congruency between odor and product may be more important during product consumption or product use than during its selection. In addition, the odor's effects may have been attenuated in our experiment because we asked our respondents to rate each product on the attribute 'has a nice smell'. PMID- 12578262 TI - Shopping center fire zone parking violators: an informal look. AB - Data for 33 1-hr. observations at a shopping center in a suburban location showed about 700 violations of a traffic regulation prohibiting parking in a fire zone. Women driving vans were the least complaint-accounting for approximately 35% of the total. PMID- 12578264 TI - Bender Gestalt Recall as a measure of short-term visual memory in children and adolescents with psychotic and other severe disorders. AB - To investigate the short-term visual memory ability of children and adolescents with severe psychiatric disorders, 82 child and adolescent inpatients and day hospital patients in a state psychiatric hospital were administered the Bender Gestalt Test as part of a psychological assessment and then asked to reproduce the designs from memory. No significant differences were found between groups on either the Bender Gestalt Recall, or the WISC-III IQs and the Digit Span and Symbol Search subtests for Psychotic Disorders (Schizophrenia, Schizoaffective Disorder, Psychosis Not Otherwise Specified), Attention Deficit Hyperactivity Disorder, Mood Disorders or Mood Disorders with co-morbid Attention Deficit Hyperactivity Disorder. The Coding subtest scores of the Psychotic Disorders group were significantly lower than the ADHD group. Analyses showed that the Bender Gestalt Recall was significantly related to age. Performance IQ, and sex. The results were discussed in terms of both the poor cognitive functioning of children and adolescents with persistent, severe mental illness, and the importance of developmental level when using the Bender Gestalt Recall as a rough measure of short-term visual memory. PMID- 12578263 TI - Relationships of objectively scored Bender variables with MMPI scores in an outpatient psychiatric population. AB - The purpose of the present study was to examine whether the Advanced Psychodiagnostic Interpretation system for the Bender Gestalt Test could reasonably predict the results of the most widely used objective measure of personality, the MMPI. Despite the widespread use of both tests, no previous studies could be found which correlated actual Bender scores with MMPI results, arising partly from the lack of a well-accepted, reliable, and objective scoring system for the Bender. The study compared the performance of 279 adult psychological outpatients on both the MMPI and Bender. The 55 Bender scorable points, which are seen most frequently in the outpatient population, were factor analyzed to yield 17 factors which were correlated with the MMPI. Significant multiple correlations were found between the Bender factors and 10 of 12 MMPI scales, with significant correlations ranging from .36 to .47. The Bender overall was able to discriminate moderately high scorers on the MMPI from low scorers. The overall results suggested that the Advanced Psychodiagnostic Interpretation scoring system includes measures that reflect general psychopathology and correlate with the MMPI as well as more specific content that is independent of the MMPI scales. The potential of this scoring system and joint use of the MMPI and Bender in personality assessment are discussed. Replication with a larger sample than 279 is encouraged for these 55 Bender and 12 MMPI items. PMID- 12578265 TI - Effect of teaching with mirrors on body image and locus of control in women college ballet dancers. AB - This preliminary study assessed the effect of mirrors in dance instruction on body image and locus of control of women college ballet dancers. With the same instructor in two beginning ballet classes 8 women were taught using mirrors and 13 women were taught without mirrors. Subjects completed the Cash 69-item Multidimensional Body-Self Relations Questionnaire, and the 40-item Adult Nowicki Strickland Locus of Control Scale during the first and last classes of a 14-wk. semester. Body-areas Satisfaction increased for the nonmirror class yet decreased for the class taught with mirrors. For the nonmirror class, there were significant increases in Appearance Orientation and Externality. Analysis suggested that the use of a mirror in the teaching of ballet may be an element which contributes to the low body-image scores of the 21 women ballet dancers. Replication with a much larger sample is required to draw conclusions. PMID- 12578266 TI - Specificity of practice in acquisition of the technique of o-soto-gari in judo. AB - Retention and transfer of the technique of o-soto-gari by 32 9-yr.-old children was optimized by conditions of acquisition similar to those of competition. PMID- 12578267 TI - Correlations for scores on conceptions of mathematics and approaches to learning mathematics. AB - This study reports research on associations of conceptions of mathematics and approaches to learning it. Scores on 63 men's and 69 women's, undergraduate students, conceptions of mathematics were associated with their approaches to learning mathematics. PMID- 12578268 TI - Misremembering the appearance of common objects: further cross-cultural confirmation. AB - The finding that people have surprisingly poor memory for the details of commonly used coins was replicated using a heretofore unstudied currency (the Irish Punt). The accuracy of 241 undergraduates in psychology in identifying a true coin from among a selection of similar fakes was low (41.9%), in line with findings from several other countries. PMID- 12578269 TI - Judgments of grammaticality of Japanese sentences lacking an argument or an adjunct. AB - This study explored Japanese speakers' linguistic intuition in judging the grammaticality of isolated simple sentences which lacked an argument or an adjunct. The argument is often required to be included in a sentence while the adjunct is not in the Japanese language, 36 participants (M age = 21.4 yr.) judged the grammaticality of (a) sentences with or without an argument or (b) sentences with or without an adjunct. Findings indicated that speakers judged the sentences without an argument as less grammatically correct than those with an argument and those with or without an adjunct. However, the grammaticality of the sentences without an argument was judged not as low as predicted by the Chomskyan linguistic analysis that adopts an all-or-none dichotomy as a criterion for grammaticality. The findings suggest that the state of actual speakers' knowledge of language differs significantly from that of idealized speakers. PMID- 12578270 TI - Students' perceptions of being graded as a group in the college classroom: relations among students' age, employment, and perceived group satisfaction. AB - This investigation compared and measured for different age groups of students, hours of employment, and previous grading experiences of one student cohort in relation to their perceived overall satisfaction with being graded as a group. A cohort of 230 students from a large southern metropolitan university enrolled in sections of two undergraduate classes. Group Interaction and Decision Making and Conflict Management, participated. Analysis indicated that (a) older students (28 47 years) were more likely to be dissatisfied with a group grade experience than middle (23-27 years) and younger (18-22 years) students. (b) Older students working part time were significantly more dissatisfied with the overall group experience than the younger part-time working students. (c) Older part-time working students were significantly more dissatisfied with the overall experience of working and being graded as a group than the middle part-time and middle full time working students. Differences were noted between the older and younger students, which supported older students' overall dissatisfaction with their experience of being graded as a group. Common complaints by older students were that younger students were immature, irresponsible, lacked "real-life" experience, and had misplaced priorities. Common complaints by younger students were that older students were too serious and rigid. It is recommended that this work be extended to include other comparison groups, graduate students, and other disciplines. PMID- 12578271 TI - Amount of photographic coverage for boys and girls on the sports page of newspapers as related to circulation size. AB - The present study analyzed the amount of photographic coverage devoted to high school athletics over 1 year. Previous research pertaining to sex differences in newspaper coverage of sports has focused on the amount of written coverage given. Present findings indicated that, as with written coverage given female athletes, photographic coverage differed significantly by newspaper circulation. Specifically, the larger the newspaper circulation, the more inequitable the photographic coverage of high school athletics was for girls. PMID- 12578272 TI - Are undergraduate examinees' perceptions of item difficulty related to item characteristics? AB - The present study assessed whether selected item characteristics--difficulty for the group, corrected item-total correlation, cognitive level, and difficulty for the examinee--relate to judgment of item difficulty. Undergraduate students in two classes (ns = 76, 43) identified what they believed were the five easiest and five most difficult items on a multiple-choice test. Statistically significant correlations were observed for difficulty for the group and the frequency that items were chosen as easy as well as with frequency of items chosen as difficult. Students performed significantly better on items they chose as easy than those chosen as difficult. Items chosen as easy more often called for simple, factual recall than did items chosen as difficult. PMID- 12578273 TI - Evaluating the phenomenon of choking in professional golfers. AB - A study involving 775 professional golfers investigated whether choking occurs in the PGA Tour's Qualifying Tournaments, known among golfers for its high pressure. It was hypothesized that players who were near or at the cutoff for earning a tour card would have higher final round scores than players whose scores entering the final round were either four or five strokes better or worse. However, the data did not support a choking hypothesis. There were no significant differences in final round scores for the conditions. PMID- 12578274 TI - Test-retest reliability of the Eurofit test battery administered to university students. AB - The purpose of this study was to examine the test-retest reliability of the Eurofit motor fitness tests performed by university students. A total of 98 undergraduate students who were enrolled in physical education departments in Greece participated (29 men aged 19.5 +/- 2.7 hr. and 66 women aged 19.4 +/- 2.7 yr.). All Eurofit motor fitness tests and anthropometric measurements were obtained twice with one week between the two measurements. Intraclass correlation coefficient indicated satisfactory coefficients above .70 for most tests. The only exception was the plate-tapping test, which yielded a low value (R = .57). Further, the majority of the Eurofit test battery fitted well within the 95% confidence interval, and only three Eurofit motor fitness test items (flamingo balance, plate tapping, and sit-ups) presented a confidence limit below the value of .70. These findings indicated that the Eurofit test battery yielded reliable data for undergraduate students. However, modifications should be considered to improve the reliability of certain test items, for application to undergraduates. PMID- 12578275 TI - Are mechanisms for perception of biological motion different from mechanisms for perception of nonbiological motion? AB - We compared the integration of information over space and time for perceiving different configurations of moving dots: a walking person (biological motion), rigid three-dimensional shapes, and unidirectional coherent motion of all dots (translation). No performance differences in judging walking direction and coherent translation direction were obtained in conditions with constant presentation times and varying number of target dots (integration over space). Depending on the speed of the two-dimensional configurations judgments were either worse or better than the judgments of walking direction. The results for conditions with different presentation times (integration over time) show that information about biological motion is integrated over time that increases with increasing gait period, while two-dimensional unidirectional motion is integrated over constant time independent of speed. The effect is not due to the oscillatory nature of the biological motion since information about a rigid three-dimensional shape is summed over a constant time independent of the period of the motion cycle. This could be interpreted as different neural mechanisms mediating the temporal summation for walking direction compared to detecting the orientation of rigid structure, or the direction of two-dimensional unidirectional motion. Since biological motion is characterized by nonrigidity, it is possible that the form itself is integrated over time and not the motion pattern. PMID- 12578276 TI - Kostmann disease--infantile genetic agranulocytosis: historical views and new aspects. AB - The results in the main reports on infantile genetic agranulocytosis or Kostmann Syndrome are summarized. New views on the pathogensis of the syndrome are given in a recent paper by Putsep et al. Kostmann syndrome may cause early onset Group B streptococcal neonatal sepsis as reported in this issue of Acta Paediatrica (9). CONCLUSION: Patients with Kostmann Syndrome who are successfully treated for agranulocytosis with serum colony stimulating factor remain deficient in cathelin LL-37, a peptide antibiotic, which is normally present in neutrophils and saliva. This deficiency may explain that patients who are successfully treated for agranulocytosis continue to suffer from oral infections such as chronic periodontitis. PMID- 12578277 TI - Growth charts for breastfed babies. PMID- 12578278 TI - "At risk" waistlines. PMID- 12578279 TI - The cognitive outcome of very preterm children: how to interpret results. PMID- 12578280 TI - Extended mother-infant skin-to-skin contact and prospect of breastfeeding. PMID- 12578281 TI - Well-presented first information supports parents' ability to cope with a chronically ill or disabled child. PMID- 12578282 TI - The joy of having a child with Down syndrome. PMID- 12578283 TI - Delayed disclosure by victims of child sexual abuse: an important topic for study. PMID- 12578284 TI - Weight reference charts for British long-term breastfed infants. AB - AIM: To produce a modification of the British 1990 weight reference reflecting the growth of long-term breastfed infants. METHODS: 120 infants from the Cambridge Infant Growth Study fed breast milk (with no formula) for at least 24 wk, with solids introduced at a mean age of 15 wk, were weighed every 4 wk from birth to 52 wk. Weights were converted to standard deviation scores (SDS) for gender and age post-term based on the revised British 1990 reference, averaged and smoothed across age, and then converted back to weights to provide modified median weight curves by gender. Other centile curves were constructed assuming the same variability and skewness by age as for the British reference. RESULTS: Long-term breastfed infants were slightly heavier than the reference at birth and crossed centiles upwards to reach +0.3 SDS at 2 mo, but subsequently crossed centiles downwards to -0.2 SDS by 12 mo. CONCLUSION: The British 1990 reference, although better than previous growth standards, reflects the growth of long-term breastfed infants only imperfectly, with mean weight falling by 0.5 SDS from 2 to 12 mo. The provision of breastfeeding-specific centiles, either as a transparent overlay or as an alternative chart, should be useful for professionals and parents to monitor the weight of long-term breastfed infants. PMID- 12578285 TI - Effect of early skin-to-skin contact after delivery on duration of breastfeeding: a prospective cohort study. AB - AIM: To study the influence on breastfeeding of skin-to-skin contact after birth. METHODS: Using a prospective cohort study design, a group of 1250 Polish children was investigated with 3 y follow-up. RESULTS: The implementation of the practice significantly increased mean duration of exclusive breastfeeding by 0.39 mo and overall breastfeeding duration by 1.43 mo. The infants kept with the mothers for at least 20 min were exclusively breastfed for 1.35 mo longer and weaned 2.10 mo later than those who had no skin-to-skin contact after delivery. The skin-to-skin contact after birth significantly coexisted with the other hospital practices supportive to breastfeeding, especially rooming-in without separation longer than 1 h per 24 h [relative risk (RR) = 3.18, 95% confidence interval (95% CI): 2.34 4.31] and first breastfeeding within 2 h after birth (RR = 2.94, 95% CI: 2.36 3.67. Multivariate analysis performed by a general linear model with duration of exclusive breastfeeding as the dependent variable indicated skin-to-skin contact and mother education as two independent variables influencing the duration of exclusive breastfeeding. CONCLUSION: The results indicate that extensive mother infant skin-to-skin contact lasting for longer than 20 min after birth increases the duration of exclusive breastfeeding. PMID- 12578286 TI - Waist circumference for the screening of the metabolic syndrome in children. AB - AIM: To identify the best anthropometric predictor of the metabolic syndrome in children. METHODS: Screening performance was evaluated in a clinical setting. The study included 140 children: 72 non-obese and 68 with non-syndromal obesity. Body mass index (BMI), waist circumference and triceps/subscapular skinfolds ratio were used as predictor variables, and systolic blood pressure, diastolic blood pressure, glucose, uric acid, fasting insulin, triglycerides and HDL-C as metabolic syndrome variables. RESULTS: The areas under the receiver operating characteristic (ROC) curves were 0.849 (95% CI: 0.780, 0.919) for BMI, 0.868 (95% CI: 0.801,0.934) for waist circumference and 0.834 (95% CI: 0.757,0.910) for the triceps/subscapular skinfolds ratio. No statistically significant differences were found for the three areas under the ROC curves. The point on the ROC curve closest to 1 corresponded to the 65th percentile for BMI, to the 70th percentile for waist, and to the 40th percentile for the triceps/subscapular skinfolds ratio. CONCLUSION: Waist circumference seems to be the best predictor of children with the metabolic syndrome in paediatric clinical settings. PMID- 12578287 TI - Lipoprotein(a) as a potent risk indicator for early cardiovascular disease. AB - AIM: Elevated levels of lipoprotein (a) [Lp(a)] are associated with increased cardiovascular risk in adults. It is not known whether Lp(a) elevation can be regarded as an additional risk factor even in children and adolescents. Therefore the purpose of this study was to compare the serum concentrations, distribution and frequency of Lp(a) and lipids of children and adolescents with premature parental and/or grandparental cardiovascular disease (CVD) with controls. METHODS: 103 children and adolescents, aged 6-18 y, from families with premature CVD in a parent and/or grandparent, i.e. before the age of 55 y, and 103 controls were estimated for lipids and Lp(a). RESULTS: Mean levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and Lp(a) were significantly higher in the risk than in the control group. Median Lp(a) levels were 20 (10 61.5) mg dl(-1) in the risk subjects, and 15 (6-26.5) mg dl(-1) in the control subjects (p = 0.005). Mean TC and LDL-C concentrations of the risk group compared with the control group were 211 mg dl(-1) (5.5 mmoll(-1)) versus 165mg dl(-1) (4.3mmol l(-1)) (p < 0.0001), and 140 mg dl(-1) (3.6 mmol l(-1)) versus 101 mg dl(-1) (2.6 mmol l(-1)) (p < 0.0001), respectively. CONCLUSION: It may be important to estimate plasma Lp(a) levels in progeny with a familial history of premature CVD, because it seems possible to identify those subjects who are at greater risk for later CVD either with or without elevated LDL-C levels. PMID- 12578289 TI - Free sialic acid storage (Salla) disease in Sweden. AB - The first 23 patients diagnosed with Salla disease in Sweden are presented. A high incidence of the "Finnish" R39C mutation, together with genealogical data, indicates that a large proportion of the mutations are of Finnish origin. All patients had pathologically high levels of free sialic acid in urine and in fibroblasts. The clinical picture confirms what has already been reported from Finland, with early psychomotor retardation, ataxia and speech problems. One third of the patients had epilepsy. CONCLUSIONS: Salla disease is more common in Sweden than supposed. A large proportion of the mutated alleles seem to be of Finnish origin. The clinical picture is the same as that reported from Finland. PMID- 12578288 TI - Abnormal heart rate response to hypercapnia in boys with an apparent life threatening event. AB - AIM: To determine instantaneous cardiac variability responses to increased carbon dioxide (CO2) during quiet sleep in infants who may be at risk for the Sudden Infant Death syndrome (SIDS). METHODS: The cardiac rate variability before, during and after a CO2 challenge was examined in 41 infants who had experienced an apparent life-threatening event (ALTE) and 41 gender- and age-matched control infants. RESULTS: The ALTE infants responded to CO2 breathing with a significant increase in R-R intervals, i.e. decreases in heart rate, compared to the controls (45.1% increase in R-R intervals vs. 41.4%; p = 0.005). The differences between ALTE infants and controls depended primarily on the boys' responses. CONCLUSION: ALTE infants, particularly ALTE boys, have an autonomic dysfunction-lower sympathetic stimulation and/or inhibited vagal withdrawal when stressed with CO2. The outcome might provide clues to the mechanisms underlying the cardiovascular processes contributing to the terminal event in SIDS. PMID- 12578290 TI - Supplementary oxygen and risk of childhood lymphatic leukaemia. AB - AIM: Childhood leukaemia has been linked to several factors, such as asphyxia and birthweight, which in turn are related to newborn resuscitation. Based on the findings from a previous study a population-based case-control study was performed to investigate the association between childhood leukaemia and exposure to supplementary oxygen and other birth-related factors. METHODS: Children born in Sweden and diagnosed with lymphatic leukaemia between 1973 and 1989 (578 cases) were individually matched by gender and date of birth to a randomly selected control. Children with Down's syndrome were excluded. Exposure data were blindly gathered from antenatal, obstetric and other standardized medical records. Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated by conditional logistic regression. RESULTS: Resuscitation with 100% oxygen with a facemask and bag immediately postpartum was significantly associated with an increased risk of childhood lymphatic leukaemia (OR = 2.57, 95% Cl 1.21-6.82). The oxygen-related risk further increased if the manual ventilation lasted for 3 min or more (OR = 3.54, 95% CI 1.16-10.80). Low Apgar scores at 1 and 5 min were associated with a non-significantly increased risk of lymphatic leukaemia. There were no associations between lymphatic leukaemia and supplementary oxygen later in the neonatal period or other birth-related factors. CONCLUSION: Resuscitation with 100% oxygen immediately postpartum is associated with childhood lymphatic leukaemia, but further studies are warranted to confirm the findings. PMID- 12578291 TI - Rhinitis in 10-year-old children and early life risk factors for its development. AB - AIM: To study the prevalence, characteristics of and risk factors for childhood rhinitis. METHODS: In a whole population birth cohort study (n = 1,456) the prevalence and characteristics of rhinitis among 10-y-old children were examined. At this age 1373 children (94%) completed standardized questionnaires, 1,043 (72%) skin-prick testing, 953 (65%) serum inhalant immunoglobulin E antibody screening and 784 (54%) methacholine bronchial challenges. RESULTS: At the age of 10 y the prevalence of hayfever ever was 18.6% and current nasal symptoms (rhinitis) 22.6%. Rhinitis at 10 y was largely seasonal and associated with low morbidity, although 62.7% of cases required pharmacological treatment. Atopy (positive skin test) and other allergic states were associated with rhinitis (p < 0.001). Wheeze or diagnosed asthma was higher with coexistent rhinitis. Among wheezing children physician-diagnosed asthma (p < 0.024) and inhaled corticosteroid use (p < 0.001) were greater with the presence of rhinitis. Significant bronchial hyperresponsiveness (methacholine concentration giving a 20% fall in forced expiratory volume in I s <4.0 mg ml(-1)) was greater if rhinitis was present even when the child did not wheeze (p < 0.001). Risk factor analysis for rhinitis identified the independent significance for atopy (p < 0.001) and eczema (p = 0.009) at the age of 4 y plus paternal rhinitis (p < 0.001), maternal rhinitis (p = 0.033) and maternal food allergy (p = 0.016). CONCLUSION: Rhinitis is common at the age of 10 y, with strong associations with atopy, wheezing, asthma and bronchial hyperresponsiveness. An inherited predisposition towards atopy appears to predominate over environment in the aetiology of this state. PMID- 12578292 TI - Measurement error associated with spinal mobility measures in children with and without low-back pain. AB - AIM: Evidence of the reliability of measurements in children is scarce, particularly in children with low-back pain. The aim of this investigation was to evaluate the measurement error associated with repeated measures of spinal mobility measures in children with and without low-back pain by establishing 95% limits of agreement. METHODS: A repeated measures study was performed involving 119 children aged 11-16 y. Of this sample 30 subjects reported recurrent low-back pain and were classified as symptomatic, the remaining 89 subjects were asymptomatic. Standardized measures were taken, including the sit-and-reach test, hip range of motion (Leighton flexometer), lumbar flexibility (modified Schober test) and lateral flexion of the spine. The same experimenter performed all testing, with 1 wk between the repeated measures. RESULTS: Correlation coefficients suggested that all measures exhibited good reliability in both the symptomatic (r = 0.80-0.95) and asymptomatic groups (r = 0.88-0.99). In contrast, the limits of agreement showed that all measures exhibited random error. The magnitude of random error was typically greater in the symptomatic subjects, suggesting that low-back pain may influence the reliability of typical measures used in this population. CONCLUSION: The magnitude of error must be interpreted in relation to analytical goals and the expected magnitude of change. In the authors' opinion the error presented appears acceptable for the serial monitoring of patients, although this will depend on the differences in mobility typically found. PMID- 12578293 TI - First information and support provided to parents of children with Down syndrome in Sweden: clinical goals and parental experiences. AB - When parents are informed that their newborn child has Down syndrome (DS), they often respond with a traumatic crisis reaction. The aims of this study were to assess the clinical goals regarding the first information and support provided to parents of newborn children with DS at the Swedish paediatric departments, and to analyse the parents' experiences of how they were first informed and treated. Data were collected during 1992-1993 from all of the 51 departments of paediatrics in Sweden. Information on the parents' experiences, collected retrospectively in 1996, was based on recollection by 165 parents of 86 children with DS born between 1989 and 1993 at 10 of the paediatric departments considered representative for Sweden. Seventy-five percent of the families were informed about the diagnosis within 24 h post partum. Some parents felt they were informed too late, and a few parents that they were told too soon. Half of the parents were satisfied with the timing. About 70% of the parents considered the information insufficient and 60% felt that they had been unsupported. Seventy percent would have liked more frequent information. Parental criticisms concerning the way in which the information was provided were that they received too much negative information about DS and that both the communication skills and the basic knowledge of DS on the part of the professionals could have been better. CONCLUSION: The Swedish paediatric departments fall short of their reported strong clinical goals regarding the initial information in Sweden, and improvements in this area are desirable. PMID- 12578294 TI - Randomized controlled trial of heparin for prevention of blockage of peripherally inserted central catheters in neonates. AB - AIM: To determine whether the addition of heparin to total parenteral nutrition (TPN) fluid would prevent blockage of peripherally inserted central catheters (PICCs) in neonates. METHODS: This was a randomized, double-blind, controlled study of 66 eligible neonates with PICCs inserted for the administration of TPN. Infants were randomized to receive TPN containing either 1 IU ml(-1) of heparin (n = 35) or no heparin (n = 31). RESULTS: There was no significant difference in the incidence of blocked catheters between the two groups of infants (heparin: 14.3%; no-heparin: 22.6%, p = 0.4). Although a higher percentage (62.9%) of infants in the heparin group received a complete course of TPN successfully via PICC than those in the no-heparin group (48.4%), the difference was not statistically significant (p = 0.3). There were no significant differences in the incidence of catheter-related sepsis, hypertriglyceridaemia, hyperbilirubinaemia, coagulopathy or intraventricular haemorrhage between the two groups. CONCLUSION: Addition of heparin to TPN fluid was not associated with a significant reduction in the incidence of blocked PICCs. However, the sample size of this study was too small to exclude even rather marked differences between the groups. PMID- 12578295 TI - Cytokine response in cerebrospinal fluid from preterm infants with posthaemorrhagic ventricular dilatation. AB - AIM: Posthaemorrhagic ventricular dilatation (PHVD) is closely associated with white matter damage and neurological disability in the preterm infant. Proinflammatory cytokines have been implicated in the pathogenesis of white matter injury and subsequent cerebral palsy. The aim of this study was to determine the levels of proinflammatory cytokines in cerebrospinal fluid (CSF) from preterm infants with PHVD and to correlate the levels to white matter damage and neurodevelopmental outcome. METHODS: CSF samples were obtained from 24 preterm infants with expanding PHVD and 19 preterm infants with normal ultrasound. Tumour necrosis factor-alphaa (TNF-alpha ), interleukin-1beta (IL 1beta), interleukin-8 (IL-8) and interferon-gamma (IFN-gamma) in CSF were measured by enzyme-linked immunosorbent assay, and IL-6 was measured by bioassay. RESULTS: The concentrations of TNF-alpha, IL-1beta, IL-6 and IL-8 were significantly elevated in CSF from infants with PHVD. TNF-alpha was detected in 43% of PHVD infants and 11% of controls (p = 0.04). IL-1beta was detected in 67% of PHVD infants and 0% of controls (p < 0.0001). The concentrations of IL-6 were 368 (145-460) pg ml(-1) in the PHVD group and 30 (25-41) pg ml(-1) in the control group (p < 0.0001), and those of IL-8 were 3000 (1620-3400) pg ml(-1) in the PHVD group and 35 (0-230) pg ml(-1) in the control group (p < 0.0001). Cytokine concentrations did not correlate with white matter lesions on ultrasound, shunt dependence or neurological outcome within the PHVD group. CONCLUSION: There was an intense and prolonged inflammatory reaction in CSF from preterm infants with PHVD and a high risk for subsequent white matter injury and permanent neurological impairment. PMID- 12578296 TI - Determinants of linear growth and predictors of severe stunting during infancy in rural Malawi. AB - Stunting is common among children under 5 y of age in sub-Saharan Africa. Several risk factors have been associated with poor growth but few studies have prospectively addressed the development of linear growth faltering and stunting during the first year of life. The present study was designed to analyse typical growth among rural Malawian infants, focusing particularly on the impact of birth size, adherence to feeding guidelines and morbidity in the development of severe stunting during infancy. A community-based cohort of 613 singleton newborns was prospectively followed by monthly home visits. Data were collected on the children's socioeconomic background, maternal size and weight gain during pregnancy, birth events, morbidity, breastfeeding and complementary feeding, growth and mortality. Univariate and multivariate analyses were used to determine associations between predictor variables and poor linear growth. The proportions of stunted infants (Height-for-age Z-score < -2) at 3, 6 and 9 mo of age were 27%, 51%, and 63%, respectively. At I y of age, over two-thirds (71%) of the infants were at least moderately (HAZ < -2) and 31% severely stunted (HAZ < -3). CONCLUSION: The strongest predictor of severe stunting at 12 mo of age was small birth size. Other variables independently associated with this outcome included inappropriate complementary feeding, high morbidity, maternal short stature, male gender, and home delivery. Faltering of linear growth started soon after birth and continued throughout infancy. Interventions increasing birth size could have a significant role in the prevention of early childhood stunting. The ideal strategy should also emphasize the importance of appropriate infant feeding and decreasing the number of illness episodes amongst the infants. PMID- 12578297 TI - Large variability in performance IQ associated with postnatal morbidity, and reduced verbal IQ among school-aged children born preterm. AB - AIM: To assess cognitive ability in a population-based group of prematurely born school-aged children and to relate these findings to postnatal morbidity. METHOD: The study group consisted of a cohort of 51 children born preterm, 43 (26 boys, 17 girls) of whom were available for psychological evaluation At evaluation, their median age was 10 y (range 8-11 y). They were all born between 1988 and 1991, with gestational age less than 29 wk (median 27, range 24-28). Their median birthweight was 1060g (range 450-1450). The Wechsler Intelligence Scale for Children (WISC-III) was used, and the test results were compared with those of a standardized, age-matched, normative group of children. RESULTS: Thirteen children (30%) performed below average [intelligence quotient (IQ) < 80] for Full Scale IQ (FSIQ). Thirty-six children had a Verbal IQ (VIQ) below the mean value of 100 [84%, 95% confidence interval 73-95%], p < 0.0001. The Performance IQ (PIQ) was within the expected range of a normal population, although a large variability was observed. Discrepancies between VIQ and PIQ of more than 15 IQ units were found in 42% of the children. High postnatal morbidity (days with assisted ventilation, number of blood transfusions) and low birthweight standard deviation scores (SDS) were associated with lower PIQ than VIQ, while low postnatal morbidity and high birthweight were associated with higher PIQ than VIQ. CONCLUSION: This cohort of preterm children had reduced overall verbal capacity independent of morbidity, and a large variability in performance capacity that was associated with postnatal morbidity. The findings suggest that there are different mechanisms influencing the outcome of verbal and performance capacity in preterm children. PMID- 12578298 TI - Risk factors for language impairment in Swedish bilingual and monolingual children relative to severity. AB - AIM: To explore potential risk factors for language impairment (LI) relative to degree of severity in bilingual and monolingual children. METHODS: Two groups of clinically assessed children were compared, 252 bilinguals whose parents were both non-Swedish and 446 monolinguals, who were referred over a period of 24 mo to the University Hospital in Malmo for suspected LI. RESULTS: A multivariate linear regression for both groups showed that two risk factors were the same: parental distress (p < 0.0001 in both groups) and short attention span (p < 0.0001 in both groups). Male gender (p < 0.0001) was an additional predictor for LI relative to degree of severity in the monolingual group. Specific risk factors for LI relative to degree of severity in the bilingual children were maternal arrival within 1 y in relation to birth (p < 0.002) and parental need for an interpreter after > 5 y in Sweden (p < 0.040). CONCLUSION: Most risk factors for LI relative to degree of severity in this study seemed to apply to both groups, although they sometimes appeared in another form in the bilingual group. Gender and possibly also hereditary factors seemed to need to interact with environmental factors to appear as risk factors for the bilingual children in this study. PMID- 12578299 TI - Expert consensus in the development of a European health-related quality of life measure for children and adolescents: a Delphi study. AB - AIM: To determine the level of consensus among experts regarding content, structure and sources of content for a new European measure of health-related quality of life in children and adolescents. METHODS: A three-round Delphi questionnaire was sent by e-mail to 24 experts in quality of life measurement in 9 European countries. Consensus was considered reached when > 90% of experts either agreed or disagreed with a given statement, or where median scores were over 6 on a scale of 1-10, and score dispersion was within predefined limits. RESULTS: Completed questionnaires were received from 20 panellists in each round. It was agreed that the new instrument should be a multidimensional, profile measure with 30-49 items covering 5-8 dimensions, which should take no more than 10-15 min to complete. Agreement was also reached on 8 specific dimensions to include in the questionnaire (psychological well-being, self-esteem, body image, cognitive functioning, mobility, energy/vitality, social relations, family/home function). Consensus was against the use of individualized questionnaires. Focus groups with children, parents and workers in the field, literature and instrument reviews were considered appropriate sources for content. CONCLUSION: Using a Delphi method in this way for the first time showed it to be a feasible and useful method for establishing a conceptual and operational framework for the Kidscreen questionnaire. PMID- 12578300 TI - Delayed disclosure and disrupted communication during forensic investigation of child sexual abuse: a study of 47 corroborated cases. AB - AIM: To study factors of relevance for the understanding of disclosure of child sexual abuse. METHODS: Cases from a Swedish district court involving 47 children in which allegations of child sexual abuse had been corroborated by a confession from the defendant were studied. RESULTS: Delayed disclosure was related to a close relationship with the perpetrator and young age at the first experience of abuse. Disrupted communication during the police interview was related to less violent abuse. CONCLUSION: The findings highlight the importance of social factors in children's disclosure of sexual abuse. PMID- 12578301 TI - Early-onset group B streptococcal sepsis in a preterm infant with Kostmann syndrome. AB - A preterm infant died of group B streptococcal sepsis 7 h after birth. The infant's complete blood count showed total agranulocytosis. Histopathology of the major organs showed significant bacterial invasion without infiltration of polymorphonuclear leucocytes. Examination of the bone marrow revealed normal cellularity of the granulocyte precursors with arrested maturation. These findings are consistent with Kostmann syndrome. CONCLUSION: It is suggested that in patients with deteriorating early-onset infection, underlying congenital abnormalities in host defence, such as Kostmann syndrome, should be considered. PMID- 12578302 TI - Neonatal cholestasis and infantile Gaucher disease: a case report. AB - AIM: To report on clinical complications of liver disease occurring during Gaucher disease. METHODS: A case of Gaucher disease was revealed by neonatal cholestasis and early onset of portal hypertension. RESULTS: At 7 d of age, a newborn was admitted for cholestasis associated with hepatosplenomegaly and thrombocytopenia. At that time, bone marrow aspirate and liver biopsy did not reveal any engorged cells. The clinical course was marked by early progressive portal hypertension, and the patient died of uncontrollable upper gastrointestinal bleeding. The histological results of the postmortem showed that Gaucher cells were present in the liver, spleen and bone marrow. The diagnosis was confirmed by enzymatic studies. CONCLUSION: Isolated neonatal cholestasis could be the first sign of Gaucher disease. Gaucher disease should always be considered in such circumstances, even if, initially, the bone marrow aspirate and liver biopsy do not reveal any engorged cells. PMID- 12578303 TI - Kikuchi Fujimoto disease: a rare presentation in a child and an updated review of the literature. PMID- 12578304 TI - Does Helicobacter pylori cause bronchiectasis? PMID- 12578305 TI - HHV 8 seroprevalence and transmission within Albanian family groups. AB - The recently discovered Human Herpesvirus 8 (HHV 8) is associated with all clinical forms of Kaposi's sarcoma. While early research suggested that the virus was transmitted sexually and that it was present only in KS patients, more recent studies seem to show that infection with the virus is more common than once thought, presenting differing distribution patterns in different geographical areas. In this study we analyze seroprevalence and transmission of HHV 8 in a sample of 86 family groups from Albania. Participants were selected among families requesting routine pre-expatriation medical examinations at the Poliambulatorio Padre "L. Monti" in Tirana. Specimens were collected from 180 healthy individuals and tested for the presence of a specific antibody. Antibody anti-HHV-8 detection was performed by immunofluorescence assay (IFA) and enzyme linked immunosorbent assay (ELISA). The study found an overall rate of HHV 8 seroprevalence of 20.0%. In 4.5% of couples the male and female were both positive, in 30.2% at least one partner was positive; (in 17.4% only the male was positive; in 12.8% only the female). These results support the hypothesis that HHV 8 spreads via multiple transmission routes. PMID- 12578306 TI - Development of a non-immunising, paraspecific vaccine from attenuated pox viruses: a new type of vaccine. AB - The various research periods leading to the development of paraspecific vaccines are described. Paraspecific vaccines are new, pyrogen-free, non-toxic preparations that contain non-immunising antigens and can be used to generate endogenic protective, non-antigen specific mechanisms in the sense of paramunization in humans and animals. They consist of highly attenuated and inactivated (0.05% Betapropiolactone) virus strains of various poxvirus genera. They activate the T helper cells and cellular elements of the paraspecific (innate) immune system and initiate the associated production and release of cytokines (cytokine cascade) with the goal of eliminating dysfunctions of the immune systems, rapidly enhancing the individual's non-pathogen- and non-antigen specific defences and exerting a regulatory effect on the interplay between the immune, hormone, nervous and vascular systems (signal-transduction mediators). They can be used systemically (intramuscularly) and locally (mucous membranes, skin). Immunization with paraspecific vaccines does not lead to postvaccinal complications and can be carried out as often as necessary, even for a number of years. They are compatible with conventional medicines and conventional specific vaccines. Closely linked protein complexes in the envelopes of the virus particles are responsible for their efficacy, some of those envelope protein complexes possess the properties of weak super antigens. Paraspecific vaccines have proved effective in combating viral infections, in particular herpes and hepatitis B and C infections, and chronic inflammatory diseases, and also as adjuvant therapy for tumours, for curing stress-related disturbances and dysfunctions of the immune system. PMID- 12578308 TI - Antimicrobial resistance of animal and human strains of Enterobacteriaceae in Greece. AB - Two hundred sixty two strains of Enterobacteriaceae from animal and human sources where intermixing and/or spreading is possible were examined for their resistance to 15 antimicrobials frequently used in animal prophylaxis and metaphylaxis. The antimicrobials with the highest proportion of resistant strains from animal sources were amoxicillin 25 mg, colistin sulphate 25 mg, erythromycin 5 mg, penicillin G 10 mg and spectinomycin 10 mg. The same with isolates from human sources where ampicillin 10 mg, amoxicillin 25 mg, colistin sulphate 25 mg, erythromycin 5 mg, neomycin 10 mg, penicillin G 10 mg and spectinomycin 10 mg. Human isolates were resistant to more antimicrobials than animal isolates. Common a high proportion of isolates from both sources were resistant to some antimicrobials. Systematic and better distributed information is needed to address the epidemiological role of veterinary treatment, prophylaxis and metaphylaxis in antimicrobial resistance in Greece. PMID- 12578307 TI - A human placental polydeoxyribonucleotide (PDRN) may promote the growth of human corneal fibroblasts and iris pigment epithelial cells in primary culture. AB - The optimal concentration of a human placental polydeoxyribonucleotide (PDRN) preparation (100 microg/ml) enhances the growth of human corneal fibroblasts in primary culture depending upon the donor age. In particular, this effect is very consistently reproducible with donors over 60 years of age (p = 0.0028), suggesting a selective benefit of PDRN in the tissue culture of senescent cells. Moreover, this drug may promote the development of human iris pigment epithelium (IPE) cells with much lower concentrations of fetal bovine serum than those suitable for the culture of IPE. Lastly, the use of a 'gauze disk' on the pieces of the corneal explants improves the efficiency of growth of the control fibroblast primary cultures. PMID- 12578309 TI - Motility of three strains of Proteus mirabilis (G9, P11 and PM5C) in liquid suspension under different environmental conditions. AB - The effect of compounds on the motility of Proteus mirabilis swarmer cells varies from one strain to another. The effect of compounds on the motility of swarmer cells is mainly at higher concentrations than the concentration used to inhibit swarming. Boric acid only affects the motility of strain G9 swarmer cells, whereas sodium deoxycholate prevented the motility of swarmer cells for three strains. Some antibiotics show their effect on the motility of swarmer cells in anaerobic areas, by slowing the movement of swarmer cells, followed by stopping the movement after a period of time or disappearance of the cells. The differentiation between the strains of Proteus species seems to be better in liquid suspension than on the solid medium. PMID- 12578310 TI - Methicillin resistance in staphylococci isolated from subclinical mastitis in sheep. AB - One hundred ovine milk samples were subjected to bacteriological analysis to detect staphylococci. Twenty-four staphylococcal strains isolated were characterised for methicillin resistance with disk diffusion test (DDT) after incubation at 24 and 48 h, oxacillin agar screen test, Minimal Inhibitory Concentration (MIC), nitrocefin test for beta-lactamase production and PCR for the mecA gene. Nine staphylococcal strains resulted resistant in DDT; some differences in the halo diameter at double incubation period were noted; eight of these strains were resistant in MIC test; just one strain was positive to oxacillin agar screen test. All strains were mecA negative by PCR and positive by nitrocefin test. On the basis of these results methicillin-resistant strains can be classified as beta-lactamase hyperproducers. PMID- 12578311 TI - Periodontal status in an Italian young adult population. Prevalence and relationship with periodontopathic bacteria. AB - To determine the prevalence of periodontitis in an Italian young adult population and the relationship with Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque. A full-mouth periodontal and oral examination was performed in 70 subjects. Dental and behaviour habits were assessed with a standardised questionnaire. Subgingival plaque samples were collected from the deepest pocket of the first molars in each quadrant with a sterile curette. A. actinomycetemcomitans, P. gingivalis and P. intermedia were detected using a multiplex polymerase chain reaction. At subject level, the prevalence of bleeding on probing, calculus, normal pocket depth (PD), PD > 5mm and bacterial positivity were 44.8%, 43.3%, 22.9%, 11.4% and 95.7%, respectively. At quadrant level bacterial prevalence was 79.4%; P. intermedia was the most common bacteria (79.0%); A. actinomycetemcomitans had a prevalence of 40.8%. A significant linear trend across categories of gingival conditions (healthy, bleeding on probing, calculus presence) was detected for P. intermedia (p = 0.0038) and A. actinomycetemcomitans (p = 0.00005) proportions. No significant association was observed between pathogenic bacteria and PD, nor with behavioural attitudes. Gingival conditions are found to be a good predictors (VPP = 85%) for periodontopathic bacteria. For the Italian population, as no data are present, prospective longitudinal studies are needed to examine the relationship between PD and bacteria presence with periodontal disease onset and/or progression. PMID- 12578312 TI - Analysis of an outbreak due to Chryseobacterium meningosepticum in a neonatal intensive care unit. AB - The aim of this study was to describe the epidemiological and clinical features of an outbreak due to Chryseobacterium meningosepticum. During a 11-day period, the outbreak was observed among four newborns in a neonatal intensive care unit (NICU) in a teaching hospital. All patients yielded C. meningosepticum in their blood cultures, in addition one was colonised in the throat. Antimicrobial susceptibility assay showed complete resistance to penicillins, cephalosporins, aminoglycosides, imipenem, aztreonam, and tetracycline, sensitivity to ciprofloxacin and trimethoprim-sulfamethoxazole. All patients were empirically treated with amikacin and meropenem. The neonate who was the first to develop sepsis died before the culture result. When C. meningosepticum was identified, antimicrobial therapy was changed to a combination of ciprofloxacin, rifampicin and vancomycin, and three neonates were treated successfully. Environmental screening recovered C. meningosepticum from two venous catheter lines and one nutritional solution that was opened by health care staff and used for two neonates. Arbitrary primed polymerase chain reaction and antibiogram typing indicated that all isolates were epidemiologically related. This study demonstrates that rapid selection of appropriate antibiotics is critical for clinical cure and standard precautions should be reconsidered to limit the spread of this bacterium on the NICU in our hospital. PMID- 12578313 TI - Serological diagnosis of brucellosis caused by Brucella canis. AB - Blood serum samples from 2,328 dogs were tested to detect antibodies against Brucella canis with the agar gel immunodiffusion (AGID) and 2-mercaptoethanol slide agglutination test (ME-SAT) using Brucella ovis as the antigen. All blood serum samples were also evaluated for antibodies against Brucella abortus and Brucella melitensis using the Rose Bengal test. Twentyfive (1.07%) of the sera evaluated were considered positive with AGID test. Only 4 (16%) of these blood serum samples were positive when evaluated with ME-SAT. The 25 AGID positive samples and 25 AGID negative serum samples were also examined by: the complement fixation test (CFT) using B. ovis hot saline extract (HSE) as the antigen, indirect enzyme linked immunosorbent assay (ELISA) and immunoblotting (IB) using B. canis and B. ovis HSE antigens. Two positive canine sera from culture positive dogs and the serum of an experimentally RM6/66 B. canis-infected rabbit were employed as positive controls and one serum from a known uninfected dog as a negative control. ELISA with B. canis antigen gave 9 (18%) positive results (6 AGID-positive and 3 AGID-negative sera). ELISA performed with B. ovis antigen detected 15 (30%) positive samples (10 AGID-positive, 5 AGID-negative and 8 B. canis ELISA positive sera). IB analysis of known positive controls sera employing B. canis antigen detected bands with molecular weights of 94-80, 64-50, 35, 32 30, 28, 23, 20-18, 15-12 kDa. The same sera tested with B. ovis antigen revealed bands of 35, 32-30, 25, 23, 20-18, 15-12 kDa. No bands were observed with the negative control serum and the 50 canine tested sera. PMID- 12578314 TI - Determination of some physiological factors affecting xylanase production from Trichoderma harzianum 1073 D3. AB - In this study, different Trichoderma strains were tested and Trichoderma harzianum 1073 D3 was found to be the most potent xylanase producer. Then some cultural parameters, namely, incubation time, substrate concentration, initial culture pH and temperature were optimized in order to increase xylanase production from Trichoderma harzianum 1073 D3. The optimum incubation time was found to be 13 days. It was concluded that 1% xylan concentration is suitable for high xylanase production rate. The optimum temperature and pH were found to be 30 degrees C and 7, respectively. Also, it was determined that agitation during growth was suitable for efficient production. PMID- 12578315 TI - Phenotyping of Cryptococcus neoformans strains in Bergamo, Italy (1985-2000). AB - Cryptococcus neoformans is the cause of the most common life-threatening fungal infection in patients with AIDS. Thirty strains of C. neoformans were collected from inpatients and typied evaluating activity, morphotyping, serotyping, chemosensitivity and adhesivity. Cryptococcus neoformans strains showed different aspectotype profile, the sole presence of serotypes A and D, good susceptibility to azoles and Amphotericin B. Phenotypic epidemiologic markers can be used: characterization of clinical strains excludes a common source. PMID- 12578316 TI - Identification of Plasmodium falciparum, P. vivax, P. ovale and P. malariae and detection of mixed infection in patients with imported malaria in Italy. AB - The species-specific nested-PCR previously described by Snounou and others for detecting the four parasite species that cause human malaria is evaluated in the current study testing 230 blood samples. The results are compared with those obtained by microscopy and, for 101 samples out of 230, with those previously obtained by a genus-specific PCR based method (pg-PCR) followed by species specific Southern-blot hybridization. All blood specimens were obtained from patients (127 foreigners and 103 Italians) with a suspect clinical diagnosis of imported malaria in Italy: 76 were positive by microscopy and 83 were positive by nested-PCR. The last method also revealed 10 double infections (8 foreigners and 2 Italians) which were not identified by microscopy or by pg-PCR with species specific Southern-blot hybridization. Fifty-four out of 83 positive samples tested by nested-PCR were submitted to genomic sequence analysis, which confirmed the presence of DNA region portion encoding the 18S rRNA corresponding to the Plasmodium species identified by nested-PCR. These results demonstrate that the nested-PCR assay surpasses microscopy and pg-PCR with species-specific Southern blot hybridization, both in sensitivity and in diagnostic accuracy. Moreover, it is quicker because it requires no further blotting or hybridization of PCR amplification products. This method also offers a clear advantage in the detection of mixed infections, which is important not only for successful medical treatment but also for the study of malaria epidemiology. Finally, our study also highlights the value of genomic sequence analysis for validating PCR results. PMID- 12578317 TI - Induced sensitization to nickel in guinea pigs immunized with mycobacteria by injection of purified protein derivative with nickel. AB - Nickel has been reported to be one of the most common causes of allergic contact dermatitis. Despite the fact that nickel is a frequent sensitizer in humans, establishing animal models for nickel allergy has met with considerable difficulties. In clinical cases, allergic contact hypersensitivity to nickel develops much more readily in inflamed skin than normal skin. In this study, we tried to induce nickel sensitization when inflammation has been evoked in guinea pigs immunized with mycobacteria followed by co-administration of a mycobacterial component with nickel. We first examined the delayed-type hypersensitivity (DTH) reaction of mycobacterial components such as the cell wall, cell membrane, 70S ribosomal fraction, cytoplasm, tuberculin purified protein derivative (PPD), RNA and DNA from Mycobacterium bovis BCG in guinea pigs immunized with live M. bovis BCG or heat killed M. tuberculosis. When PPD was used, the hypersensitivity reaction was strongest. Next, we tested whether PPD with nickel could induce nickel sensitivity in guinea pigs immunized with mycobacteria. Strong sensitization to nickel was achieved by injecting PPD with nickel. However, if too large an amount of PPD or nickel salts was used, sensitization to nickel decreased. In this way, sensitization of nickel developed much more easily in guinea pigs immunized with mycobacteria by injection of an appropriate amount of nickel at the inflammation site induced by a suitable amount of PPD. PMID- 12578318 TI - Typing of Pseudomonas aeruginosa strains in Turkish cystic fibrosis patients. AB - The majority of cystic fibrosis (CF) patients suffer from chronic respiratory infection with the opportunistic bacterial pathogen Pseudomonas aeruginosa. The virulence of P. aeruginosa is associated with the presence of various extracellular factors, like alginate, elastase, alkaline protease which contribute tissue destruction and assist bacterial invasion. Virulence factor production of P. aeruginosa strains isolated from 46 CF patients followed in two cities in Turkey was detected. Strains were compared genotypically by arbitrarily primed PCR. Antimicrobial susceptibilities to 12 antibiotics were determined by broth microdilution method. Evaluation of virulence factor results revealed that 95.8% of the strains were alginate, 71.7% elastase and 52.1% alkaline protease producers. AP-PCR analysis revealed 35 genotypes indicated almost a complete discrepancy among the strains. The most effective drugs were penems and quinolones. Among aminoglycosides amikacin was the most effective one and a high level resistance to beta lactams was observed. Alginate is the most important virulence factor in the chronic colonisation of CF patients with P. aeruginosa. No evidence for cross infection between patients and for relationship between phenotypes and genotypes of the strains was found. PMID- 12578319 TI - Inhibitory activity of spices and essential oils on psychrotrophic bacteria. AB - This study was designed to evaluate "in vitro" the inhibitory effects of spices and essential oils on the growth of psycrotrophic food-borne bacteria: Aeromonas hydrophila, Listeria monocytogenes and Yersinia enterocolitica. The sensitivity to nine spices and their oils (chilli, cinnamon, cloves, ginger, nutmeg, oregano, rosemary, sage, thyme) was studied. Antibacterial activity was evaluated on liquid and solid medium. Spices: 1% concentration of each spice was added separately to Triptic Soy Broth and then inoculated to contain 10(8)/ml organism and held to 4 degrees C for 7 days. Populations of test organism were determined on Triptic Soy Agar. Oils: Inhibition of growth was tested by using the paper disc agar diffusion method (at 35, 20 and 4 degrees C) and measuring their inhibition zone. MIC was determined by the broth microdilution method. Some culinary spices produce antibacterial activity: inhibition of growth ranged from complete (cinnamon and cloves against A. hydrophila) to no inhibition. Antibacterial inhibition zone ranged from 8 mm to 45 mm: thyme essential oil showed the greatest inhibition against A. hydrophila. PMID- 12578320 TI - Molecular approaches to the identification and treatment monitoring of periodontal pathogens. AB - Two different PCR-based molecular approaches, a commercial kit for detection of A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus and T. denticola (Amplimedical "Paradonthosis") and a home-made multiplex PCR for A. actinomycetemcomitans, P. gingivalis and B. forsythus were compared for monitoring the efficacy of different dental treatments on localized persistent periodontal pockets. 44 sites were randomized in two treatment groups: mechanical treatment (22 control sites) and in conjunction with the application of tetracycline fibres (22 experimental sites). 40/44 sites were found positive with both tests for A. actinomycetemcomitans, P. gingivalis and B. forsythus pretheraphy. P. intermedia was detected alone in only three sites during the follow-up, while T. denticola. was always associated with the other pathogens. 20 sites were positive in conventional cultures for one to three of the pathogens. PCR-based approaches provided a sensitive and reliable method for identification and monitoring treatment of periodontal pathogens. PMID- 12578321 TI - Significance of inoculum size in biofilm formation by staphylococci. AB - In the present study the effect of inoculum size, ranging from 10(6) to 1 cell, in biofilm formation by staphylococci was determined by microtiter plate test. The initial inoculum size had a dramatic effect on the quantity of biofilm formed. A decreased number of bacteria in initial inoculum always resulted in a decreased production of biofilm. PMID- 12578322 TI - Neurologic impairment in children associated with maternal dietary deficiency of cobalamin--Georgia, 2001. AB - During 2001, neurologic impairment resulting from cobalamin (vitamin B12) deficiency was diagnosed in two children in Georgia. The children were breastfed by mothers who followed vegetarian diets. This report summarizes the two cases and provides guidance for health-care providers on identifying and preventing cobalamin deficiency among breastfed infants of vegetarian mothers. PMID- 12578323 TI - Pneumococcal conjunctivitis at an elementary school--Maine, September 20-December 6, 2002. AB - On October 18, 2002, the nurse at an elementary school in Westbrook, Maine, notified the Maine Bureau of Health (MBOH) of an increase in the number of students with conjunctivitis. During September 23-October 18, a total of 31 students in kindergarten and in first and second grades either were reported by parents to the nurse as having conjunctivitis or had conjunctivitis diagnosed by the nurse at school. Conjunctival swab cultures from five (38%) of the 13 students who were tested initially grew Streptococcus pneumoniae. This report documents additional cases in the community and summarizes preliminary results of the investigation of this outbreak, which indicated that the outbreak was caused by the same nontypeable strain of pneumococcus that caused an outbreak of conjunctivitis among college students in New Hampshire during January-March 2002. This is the first time that this strain has been reported as the cause of a conjunctivitis outbreak among schoolchildren. Health-care providers and public health officials should be aware that nontypeable S. pneumoniae can cause outbreaks of conjunctivitis in school-age children and college students; outbreaks should be reported to state health departments and CDC. PMID- 12578324 TI - Lower extremity amputation episodes among persons with diabetes--New Mexico, 2000. AB - Lower extremity amputation (LEA) is one of the most disabling complications of diabetes. Lower extremity problems tend to recur among persons because of underlying complications, including the loss of "protective" sensation. To define the burden of LEA among persons with diabetes in New Mexico, the New Mexico Diabetes Prevention and Control Program (DPCP) analyzed data from the Hospital Inpatient Discharge Database (HIDD) and the Santa Fe Indian Hospital (SFIH) from 2000 by linking hospital discharges to persons to create "episodes" of LEA. This report summarizes the findings of that analysis, which indicated that the age adjusted rate of LEA by episode was approximately 3.5 times higher for American Indians (AIs) (11.4 per 1,000 persons with diabetes) than for non-Hispanic whites (3.3). To address this disparity, DPCP is collaborating with the Indian Health Service (IHS) to determine the needs for foot-care resources and education in AI communities. PMID- 12578325 TI - Facilitating influenza and pneumococcal vaccination through standing orders programs. AB - Influenza and pneumococcal vaccines are underused for persons in the United States aged > or = 65 years (66% receive influenza vaccine and 55% pneumococcal vaccine), even among patients in nursing homes (68% for influenza and 38% for pneumococcal vaccine). Systematic literature reviews by the Task Force on Community Preventive Services and the Southern California Evidence-Based Practice Center-RAND have shown that standing orders programs improve vaccination rates. Standing orders programs authorize nurses and pharmacists, where allowed by state law, to administer vaccinations according to an institution- or physician approved protocol without the need for a physician's examination or direct order. Several studies have shown improved influenza and pneumococcal vaccination rates through standing orders programs specifically in long-term care facilities (LTCFs) and hospitals. Based on the strength of available evidence, the Advisory Committee on Immunization Practices recommends the use of standing orders programs in both outpatient and inpatient settings. PMID- 12578326 TI - Recommended childhood and adolescent immunization schedule. United States, 2003. Centers for Disease Control and Prevention. AB - Each year, CDC's Advisory Committee on Immunization Practices (ACIP) reviews the recommended childhood and adolescent immunization schedule to ensure that it is current with changes in manufacturers' vaccine formulations and contains revised recommendations for the use of licensed vaccines, including those newly licensed. The recommended childhood immunization schedule for 2003 has remained the same in content and format since January 2002 (Figure 1). The recommendations and format have been approved by ACIP, the American Academy of Family Physicians, and the American Academy of Pediatrics. PMID- 12578327 TI - Regional differences in surgical heart valve disease in Europe: comparison between northern and southern subsets of the EuroSCORE database. AB - BACKGROUND AND AIMS OF THE STUDY: Although rates for coronary and valve surgery vary between northern and southern Europe, differences in the features of valve disease leading to surgery in Europe are poorly documented. The study aim was to compare demographics, risk factors, procedures and outcome in valve surgery between European regions, using the EuroSCORE database. METHODS: Between September and December 1995, information on 98 variables (risk factors, procedures and outcome) were collected on valve surgery patients in 128 European centers. Patients were allocated to two geographic subgroups (north, n = 1,990; south, n = 3,682). The distribution of variables was assessed. Subsequently, the impact of preoperative and operative risk factors on mortality was analyzed in both groups using a bivariate analysis. Risk-adjusted outcomes were then compared according to the EuroSCORE. RESULTS: Significant differences were identified for clinical features, risk factors and procedures. In northern Europe, surgery was performed on older patients with more severe coronary or associated disease, whilst in the south the cardiac status seemed more severely compromised. Degenerative aortic disease prevailed in the north (aortic valve replacement in 72.7% of cases), whilst in the south mitral surgery accounted for 46.1% of procedures. Despite differences in crude mortality (6.9% north versus 5.7% south), outcomes (when adjusted to risks) seemed comparable (observed-to-expected mortality ratio 0.90 for north versus 0.84 for south). The impact of individual risk factors on mortality was similar, except for atrial fibrillation. CONCLUSION: Despite large epidemiological differences between northern and southern Europe in terms of valve surgery, performances and outcomes were similar when individual risk factors and overall risk profiles were taken into account. PMID- 12578328 TI - Angiotensin-converting enzyme genotype predicts valve damage in acute rheumatic fever. AB - BACKGROUND AND AIM OF THE STUDY: The genetic basis for host susceptibility to subsequent valve damage and scarring is not well defined in patients with a history of acute rheumatic fever (ARF). A high angiotensin-converting enzyme (ACE) activity has been demonstrated in valve tissue; hence, the study aim was to determine whether the ACE-DD genotype was a predisposing factor to heart valve damage after ARF attack. METHODS: A total of 165 patients diagnosed previously (16 +/- 5 years ago) with ARF was evaluated. Diagnoses were conducted according to Jones' criteria, and all received similar treatment and remain on regular penicillin prophylaxis. The ACE genotype was determined using polymerase chain reaction methods. RESULTS: Echocardiography showed that 39 patients (11 males, 28 females; mean age 25 +/- 6 years) had completely normal valves, and 126 patients (38 males, 88 females; mean age 21 +/- 6 years) had valve disease. The mitral valve was involved in 93% of patients (stenosis 86%, regurgitation 69%), and the aortic valve in 50% (stenosis 19%, regurgitation 48%). The ACE-DD genotype is associated with a significantly greater risk of valve involvement (risk ratio 2.7, 95% CI 1.15-6.5, p = 0.02). The distribution of genotypes was similar between aortic and mitral valve involvement. CONCLUSION: In patients with acute rheumatic fever the ACE-DD genotype is associated with an increased risk of subsequent heart valve damage. PMID- 12578329 TI - Assumptions and realities of mitral valve repair. PMID- 12578330 TI - Trends in mitral valve surgery in a large multi-surgeon, multi-hospital practice, 1979-1999. AB - BACKGROUND AND AIM OF THE STUDY: Trends in mitral valve operations (MVO) may help to predict the future of mitral valve surgery in the context of changing case mix, population demographics, emerging technology and shifting paradigms. METHODS: All adults undergoing single mitral valve operations (MVO) between 1979 and 1999 were reviewed retrospectively according to age, gender and other typical clinical variables, surgical complexity, specific operation and immediate outcome. RESULTS: A total of 2,055 single MVO was performed. Although coronary artery bypass volumes declined by 15.3% from 1996 to 1999, MVO volumes have continued to increase 58.0% since 1996. For the entire period, there was an insignificant increase in mean age, but an increase in percent male gender and in the prevalence of degenerative and ischemic etiology and mitral regurgitation (MR) pathophysiology. During the 1990s, trends in surgical complexity included a stable 40% prevalence of combined MVO and a stable 9:1 distribution of first operations to reoperations. Technology adoption included a decreased prevalence of mechanical valve usage at the expense of an increased prevalence of mitral valve repair (MVRpr). The prevalence of MVRpr among individual surgeons appeared to be related to a threshold case load of 20 mitral valve operations per year. Predictors of hospital mortality rates for MVO included age > or = 65 years, reoperations and combined MVO. Age > or = 65 years was a predictor of hospital mortality for each category of overall, isolated and combined MVO, mitral valve replacement (MVR) and MVRpr except for combined MVR. Hospital mortality rates for overall MVO, first-op MVO and combined MVO decreased during the 1990s. Hospital mortality comparisons between MVR and MVRpr favored MVRpr, either significantly or by insignificant trend, in every category of overall, isolated and combined groups and when evaluated by age > or = or < 65 year, in overall, isolated and combined groups. CONCLUSION: MVO volumes are steadily increasing apparently as a result of the increase in octogenarians and the beginning of the 'baby boomer' wave. Degenerative and ischemic etiologies with MR pathophysiology are on the rise, while rheumatic and endocarditis etiologies are static. The prevalence of MVR with mechanical prostheses has decreased in favor of MVRpr. The prevalence of MVRpr among individual surgeons appears to be related to an annual threshold volume of overall MVO. Hospital mortality risk is related to age and surgical complexity, but is modest and has continued to trend down during the past decade. Hospital mortality appears to favor MVRpr over MVR in all categorical comparisons, either significantly or by insignificant trend. These opposite trend lines for MVR and MVRpr likely represent a paradigm shift away from mechanical solutions in favor of tissue solutions for mitral valve disease, especially for MVRpr. PMID- 12578331 TI - Mitral valve replacement in children. AB - BACKGROUND AND AIMS OF THE STUDY: Although repair of the mitral valve in children with or without concomitant congenital heart defects has improved significantly, it is not always achievable. The study aim was to review a 20-year experience of mitral valve replacement (MVR) in children. METHODS: Since 1980, 90 patients (37 males, 53 females; mean age 8.1 years; range: 3 weeks to 18 years) have undergone a total of 102 MVR operations (12 redo-MVR, nine multiple valves, and three with common atrioventricular valve replacement). Tissue valves were used in 13 patients (14%). The etiology for valve disease was congenital in 72 patients (80%), and 34 patients had atrioventricular septal defect (AVSD). Other etiologies included rheumatic heart disease (n = 8), myxomatous disease (n = 4), endocarditis (n = 3), and Kawasaki disease, left atrial myxoma and idiopathic hypertropic subaortic stenosis (each n = 1). In total, 36 patients (40%) had a previous mitral valve repair, and 34 (38%) had concomitant repair of associated lesions. RESULTS: Hospital mortality was significantly higher in children aged < 2 years (52%, 15 of 29) compared with older children (3%, 2 of 61) (p < 0.001). Fourteen hospital deaths were associated with failed repair of complex congenital heart defects, mainly AVSD under age 2 years, followed by MVR. Mean follow up was 9.3 years (range: 7 months to 21.5 years). There were four late deaths; major events included thromboemboli (n = 6), bleeding (n = 9), endocarditis (n = 1) and cardiomyopathy with orthotopic heart transplantation (n = 7). CONCLUSION: MVR is a good surgical option for a nonrepairable mitral valve in children aged over 2 years. MVR following failed AVSD repair carries a high incidence of morbidity and mortality. PMID- 12578332 TI - Effect of atrial fibrillation on the dynamics of mitral annular area. AB - BACKGROUND AND AIMS OF THE STUDY: The mitral annulus shows dynamic changes in shape and size during the cardiac cycle. A smaller size in end-diastole is attributed to the sphincteric action of atrial systole, and this may be important for functional integrity of the mitral valve. However, the effect of atrial fibrillation (AF) on dynamic changes in mitral annular size in humans is not known. METHODS: Mitral annular diameters in apical four- and two-chamber views were measured using echocardiography in 25 patients in atrial fibrillation, and in 37 subjects in normal sinus rhythm at mid-diastole, end-diastole and end systole. Mitral annular area was computed assuming elliptical geometry. RESULTS: Patients in sinus rhythm showed a significant increase in mitral annular area of 25.9 +/- 12.8% with ventricular systole compared to its area in end-diastole (p < 0.0001), and a 10.5 +/- 8.4% reduction with atrial systole compared to mid diastole (p < 0.001). Patients in AF had larger mitral annuli which showed non significant changes in size between these three phases of the cardiac cycle. Percent reduction in mitral annular area in the latter half of diastole correlated significantly with left atrial (LA) diameter (r = -0.54, p < 0.0001), LA volume (r = -0.50, p < 0.0001), left ventricular (LV) fractional shortening (r = 0.37, p = 0.0036), mitral annular area in mid-diastole (r = -0.41, p = 0.0011) and mitral annular area in end-diastole (r = -0.64, p < 0.0001). That is, atrial sphincteric action on the mitral annulus was less in the presence of larger left atrium or the mitral annulus. Stepwise multiple regression analysis showed rhythm and mitral annular size to be independent predictors of dynamic changes in mitral annular area. CONCLUSION: It is concluded that AF blunts or eliminates the phasic changes in mitral annular size during the cardiac cycle with loss of its presystolic sphincteric action; this may have implications in the genesis and surgical correction of mitral regurgitation. PMID- 12578333 TI - Lack of association of genetic polymorphisms in the interleukin-1beta, interleukin-1 receptor antagonist, interleukin-4 and interleukin-10 genes with mitral valve prolapse in Taiwan Chinese. AB - BACKGROUND AND AIMS OF THE STUDY: Inflammation and genetics may play a role in the pathogenesis of mitral valve prolapse (MVP). The study aim was to test whether interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL-1Ra), IL-4 or IL-10 gene polymorphisms could be used as markers of susceptibility or severity in MVP among the Chinese population in Taiwan. METHODS: A group of 100 patients with MVP diagnosed echocardiographically, and 103 age- and sex-matched normal control subjects was studied. IL-1beta promoter, IL-1beta exon 5, IL-1Ra, IL-4 promoter, IL-4 intron 3 and IL-10 gene polymorphisms were identified by polymerase chain reaction-based restriction analysis. RESULTS: There was no significant difference in the distribution of genotypes and allelic frequencies between MVP cases and controls for IL-1beta promoter, IL-1beta exon 5, IL-1Ra, IL 4 promoter, IL-4 intron 3 and IL-10 gene polymorphisms. Further categorization of MVP patients into mild and severe subgroups also revealed no statistical difference in these gene polymorphisms when compared with controls. CONCLUSION: These findings suggest that the IL-1beta, IL-1Ra, IL-4 or IL-10 gene polymorphisms are not suitable genetic markers of MVP in Taiwan Chinese. PMID- 12578334 TI - Mitral valve re-replacement for impaired bioprosthesis after 19 years in a patient undergoing steroid treatment. AB - A 59-year-old man underwent closed mitral commissurotomy for mitral stenosis at the age of 19 years, and mitral valve replacement (MVR) with a Hancock standard bioprosthesis and tricuspid annuloplasty at the age of 40 years. Two years after the second operation, the patient developed human T-lymphotropic virus type-1 infection, manifesting as myelopathy/tropical spastic paraparesis, and has required steroid therapy for 17 years. He was well for 18 years after the MVR, but has recently developed severe congestive heart failure, orthopnea and enlarged liver. Doppler echocardiography showed severe transvalvular regurgitation of the prosthesis and tricuspid valve. The patient underwent re-MVR with a 27 mm St. Jude Medical prosthesis, and tricuspid annuloplasty. The surgery was well tolerated, without any significant infectious complication. Some tears and perforations were found in the leaflets of the explanted bioprosthesis, but soft X-ray analysis showed minimal calcification. Mild changes and longer durability of the bioprosthesis may be related to the long-term use of steroid therapy. PMID- 12578335 TI - Huge prosthetic mitral valve thrombus: use of serial transesophageal echocardiography guidance in resolution. AB - The case is presented of a 35-year-old woman with major thrombotic obstruction of a 29-mm St. Jude Medical mitral valve prosthesis. The thrombi were treated with successive intravenous fibrinolysis under transesophageal echocardiographic guidance, without complication. One large and obstructive thrombus with a long and mobile component, and other smaller and mobile thrombi were detected. These were gradually lysed after two successive administrations of streptokinase (1.5 x 10(6) units over 24 h), and one session of recombinant tissue type plasminogen inhibitor (100 mg over 6 h). PMID- 12578336 TI - Bicuspid versus tricuspid aortic valves. PMID- 12578337 TI - Histopathological grading of ascending aortic aneurysm: comparison of patients with bicuspid versus tricuspid aortic valve. AB - BACKGROUND AND AIMS OF THE STUDY: Bicuspid aortic valve (BAV) is a common inherited condition that is often accompanied by ascending aortic aneurysm. A high level of histological wall abnormalities was reported to be present in non dilated aortas of patients with BAV. In patients with tricuspid aortic valve, there appears to exist a direct relationship between the diameter of the ascending aorta and degree of histopathological aortic wall abnormalities. Whether this situation exists in patients with BAV has not yet been investigated. METHODS: Surgical and medical records of all patients undergoing surgery of the ascending aorta were reviewed. A total of 65 patients was identified in whom an aortic wall specimen was obtained intraoperatively. These specimens were systematically re-evaluated, and graded according to the severity of seven histopathological conditions: fibrosis, atherosclerosis, medionecrosis, cystic medial necrosis, smooth muscle cell orientation, elastic fiber fragmentation, and inflammation. RESULTS: BAVs were present in 26 patients (40%). Patients with BAV had significantly less aortic wall alterations than patients with tricuspid aortic valves (p < 0.001) in all variables examined. The severity of aortic wall abnormalities was significantly dependent on aortic diameter in patients with BAV as well as tricuspid aortic valve (p = 0.036 and 0.019), but dependent on age (p = 0.009) only in patients with tricuspid aortic valve. CONCLUSION: The study results provide evidence that ascending aortic aneurysm in patients with BAV differs clinically and histologically from that in patients with tricuspid aortic valve. Further studies are needed to elucidate the impact of inherited and acquired aortic wall abnormalities on the development of aneurysms. PMID- 12578338 TI - Valve-preserving aortic root reconstruction: a comparison of techniques. AB - BACKGROUND AND AIM OF THE STUDY: Controversy exists with regard to the indications and technique for valve-preserving aortic root reconstruction. Here, the authors' clinical experience with three techniques is reported. METHODS: Between October 1994 and October 2000, 71 patients (46 men, 25 women; median age 53 years; range: 21-81 years) underwent aortic root reconstruction with preservation of the aortic valve and replacement of ascending aorta. Factors that affected survival and need for subsequent aortic valve replacement were analyzed using univariate analysis. RESULTS: Indications for operation were chronic aneurysm in 65 patients, acute dissection in five, and chronic aneurysm with dissection in one patient. Forty patients were asymptomatic. Presenting symptoms included shortness of breath in 16 patients, chest pain in 14 and transient ischemic attacks in one patient. Twenty patients had Marfan syndrome. The reimplantation technique was performed in 52 patients, remodeling technique in 14, and remodeling with sinus graft root replacement technique in five. Postoperative echocardiography demonstrated absent to mild aortic insufficiency in 63 patients. Operative mortality was 2.8%. Actuarial survival was 84.1% at 5 years; freedom from reoperation was 88.8% and 78.9% at one and three years, respectively. Eleven patients (21.2%) repaired by the implantation technique required aortic valve replacement secondary to aortic valve insufficiency. By univariate analysis, the necessity of aortic cusp repair (p = 0.0009), aortic annulus size >25 mm (p = 0.04), and male gender (p = 0.04) were predictors for subsequent aortic valve replacement. CONCLUSION: Aortic valve-preserving surgery is possible with a low morbidity and mortality. The authors' experience shows a high failure rate in males with a large annulus needing aortic cusp repair and undergoing the reimplantation technique. PMID- 12578339 TI - Persistence of Chlamydia pneumoniae in degenerative aortic valve stenosis indicated by heat shock protein 60 homologues. AB - BACKGROUND AND AIMS OF THE STUDY: Based on the concept of chronic persistent infections with Chlamydia pneumoniae among variable stressors for aortic valve degeneration, the study aim was to assess the presence of chlamydial heat shock protein (cHSP) 60 and its human homologue (hHSP60) in diseased valvular tissue. METHODS: Surgical specimens of high-grade stenosed, native (n = 33) and bioprosthetic (n = 10) aortic valves were examined immunohistochemically for the localization of cHSP60, hHSP60 and macrophages (CD68), supplemented by polymerase chain reaction (PCR) and electron microscopy to prove microbial presence. RESULTS: Degenerated valves showed specific immunostaining of cHSP60 in 27 cases (65%), of hHSP60 in 26 (63%), and of CD68 in 36 (84%). Both HSP60 homologues were predominantly detected in valvular fibrosa, consistently co-localized with macrophages and, quantitatively, showed a strong correlation (r = 0.81, p < 0.001). Presence of C. pneumoniae was demonstrated by PCR in a subset of 11 of 18 valves (61%). Microbial persistence was confirmed by ultrastructural analysis. Degenerated prosthetic valves revealed markedly higher macrophage infiltration and cHSP60 signaling compared with degenerated native valves (each p < 0.05). CONCLUSION: Beyond detection of C. pneumoniae, the present data on co localization and valvular predilection sites (fibrosa) of both HSP60 homologues indicate the presence of chronic persistent C. pneumoniae infection as well as regional stressor effects, and suggest their involvement in native and prosthetic valve degeneration. PMID- 12578340 TI - Minimally invasive versus conventional aortic valve replacement: a prospective randomized trial. AB - BACKGROUND AND AIM OF THE STUDY: A prospective randomized study was performed to compare conventional with minimally invasive aortic valve replacement (AVR). METHODS: Forty consecutive patients scheduled for elective aortic valve surgery were prospectively randomized either to the conventional group (group A, complete median sternotomy) or minimally invasive group (group B, partial upper sternotomy). Intraoperative and postoperative clinical data, and markers of myocardial and cerebral protection were determined. Neuropsychological tests were carried out to quantify psychological disorders. RESULTS: Operative time and cardiopulmonary bypass time were slightly longer in group B, but not significantly so. No significant inter-group differences were found for postoperative pain scores and respiratory function. Chest tube drainage was significantly less in group B (495 +/- 165 versus 240 +/- 69 ml, p = 0.008). Creatine kinase (CK), CK-MB and troponin T levels were similar in both groups. Neither S-100B protein nor neuron-specific enolase levels differed significantly between groups at all sampling times. There were no strokes in the entire cohort. None of the neuropsychological tests yielded significant inter-group differences between conventional and minimally invasive surgery. CONCLUSION: The safety and reliability of AVR via a partial upper sternotomy is reported. Minimally invasive AVR can be performed with only slightly longer operative times, good cosmetic results and significantly less blood loss. A limited surgical access affected neither the patients' neurological outcome nor the efficacy of myocardial protection. PMID- 12578341 TI - Fourteen years' experience with the CarboMedics valve in young adults with aortic valve disease. AB - BACKGROUND AND AIM OF THE STUDY: Homografts and the Ross procedure are recommended by many surgeons for aortic valve replacement (AVR) in young adults. The study aim was to monitor patient outcome over a 14-year period after implanting mechanical CarboMedics prosthetic heart valves in young adults with aortic valve disease. METHODS: Between November 1987 and December 2000, 55 patients (42 males, 13 females; median age 33 years; range 15-40 years) each received a CarboMedics valve in the aortic position. All patients were included on a consecutive and non-selected basis. Concomitant surgery was performed in five patients. Anticoagulation was commenced on postoperative day 1. After discharge, patients were followed up by their cardiologist or general practitioner using questionnaires and telephone calls. Actuarial estimates of survival and freedom from morbid events were calculated using the Kaplan-Meier method. RESULTS: No patient died during surgery. Complete follow up data were available for all patients; total follow up was 404 patient-years (pt-yr). The 30 day mortality rate was zero. Actuarial survival at 14 years was 90 +/- 4.9%, and actuarial freedom from all valve-related events 92 +/- 3.7%. No valve thrombosis or major bleeding events were identified. Linearized rates per 100 pt-yr were: embolism 0.25; paravalvular leakage 0.25; prosthetic endocarditis 0.25; and reoperation 0.50. There was no hemolysis, prosthetic valve dysfunction, or structural deterioration. CONCLUSION: Over a 14-year period, implantation of the CarboMedics prosthetic heart valve has proven to be an excellent solution for AVR in young adults with aortic valve disease. PMID- 12578342 TI - Cellular expression of PCNA and procollagen in vital and ethanol-treated autologous pericardial implants in sheep. AB - BACKGROUND AND AIMS OF THE STUDY: Cardiovascular surgeries involving repair or reconstruction of heart valve leaflets with vital autologous pericardium have shown detrimental healing outcomes, mainly fibrosis with retraction. It is proposed that cells intrinsic to the pericardial implants may contribute to this fibrosis by becoming activated to proliferate and synthesize type I collagen. METHODS: Vital and ethanol-treated autologous pericardium were implanted as rectangular flaps bisecting the lumen in the descending aorta of sheep to simulate a heart valve leaflet. Implants recovered at 5, 10, 15, and 30 days were evaluated immunohistologically for expression of PCNA and procollagen. RESULTS: In ethanol-treated pericardium, concentrations of activated cells shifted from the fibrin layers on the periphery of implants at days 5 and 10 to cells internal to the implant at days 15 and 30. In contrast, concentrations of activated cells in vital pericardium shifted from cells within the implants at days 5 and 10 to the fibrin deposits overlaying the implants at days 15 and 30. CONCLUSION: Different distributional patterns of activated cells were observed between vital and ethanol-treated pericardial flap implants. These different patterns may be important in understanding the cause of the detrimental healing outcome observed with vital autologous pericardial flap implants. PMID- 12578343 TI - Detoxification on top of enhanced, diamine-extended glutaraldehyde fixation significantly reduces bioprosthetic root calcification in the sheep model. AB - BACKGROUND AND AIM OF THE STUDY: Increased concentrations of glutaraldehyde (GA), diamine-extension (DA) of crosslinks and subsequent extraction of excess GA all reduce bioprosthetic calcification in the subdermal rat model. The study aim was to demonstrate the combined effect of all three treatments in a circulatory sheep model. METHODS: Two fixation treatments were used for GA detoxification (urazole in acetate buffer, 0.1 M; pH 4.5; 37 degrees C; 7 days): (i) conventional 0.2% GA fixation (4 degrees C; 7 days); and (ii) enhanced 3.0% GA fixation (4 degrees C; 2 days, followed by a DA interim step; 100 mM L-lysine; 37 degrees C; 2 days, followed by GA; 3.0%; 37 degrees C; 5 days). Entire porcine root prostheses were implanted in the distal aortic arch of young sheep for 12 weeks (n = 5 per group). Non-detoxified 0.2% GA-treated roots served as controls (n = 5). Calcium analysis was based on atomic absorption spectrophotometry; morphology was assessed using light and transmission electron microscopy. RESULTS: Detoxification alone resulted in an 83% reduction of leaflet calcification (p = 0.086), but achieved only 23% (p = 0.145) and 12% (p = 0.362) mitigation of calcification in aortic wall and sinus tissue, respectively. When combined with DA-enhanced 3% GA fixation, detoxification led to a 95% reduction in leaflet calcification (p = 0.057), followed by 79% in sinus (p = 0.003) and 79% in aortic wall tissue (p = 0.0003). Morphologically, detoxification primarily affected leaflets and the subadventitial layer of aortic wall tissue, whereas enhanced fixation seemed to affect all structures. CONCLUSION: It was shown in a circulatory sheep model that a combination of DA-enhanced fixation with an extraction process of excess GA leads to a distinct mitigation of leaflet and aortic wall calcification. PMID- 12578344 TI - Dynamic impact stress analysis of a bileaflet mechanical heart valve. AB - BACKGROUND AND AIMS OF THE STUDY: Mechanical heart valves (MHV) are widely used to replace dysfunctional and failed heart valves. The bileaflet MHV is very popular due to its superior hemodynamics. At present, bileaflet MHVs account for about two-thirds of the prosthetic heart valve market. Since their introduction in 1977, the hemodynamics of bileaflet prostheses has been extensively studied. New technologies used to develop MHV include better design concepts, materials, manufacturing processes, and post-design verification. The study aim was to investigate the dynamic impact stress of a newly designed bileaflet MHV under normal physiological conditions. METHODS: Pro/Engineer was used to generate a 3-D model of the designed valve. ANSYS 5.5 and LS-DYNA were used to calculate stress and deformation of the valve. Due to symmetry, a one-half orifice and one leaflet were modeled using the eight-noded hexahedral elements. When valve leaflets are in the fully closed position, the static contact stress between leaflet and orifice was predicated under typical heart valve closing pressure of 80 mmHg. To study the dynamic effects of the closing valve, LS-DYNA was used to simulate leaflet motion. Typical physiological pressure waveform was employed to initiate this leaflet motion. Two types of valve were investigated: Test valve A (size 19, flat leaflet); and test valve B (size 19, tapered leaflet 1.5 degrees, with the same thickness at pivot as valve A). The non-invasive laser sweeping technique was used to measure leaflet closing velocity in a mock flow test rig. The closing velocity of test valve A was compared by experimental and computed results. The corresponding dynamic contact stress on the leaflet was obtained for different modes of loading, simulated under angular velocity, acceleration, and especially under representative pressure waveform. RESULTS: The experimental closing velocity of test valve A was 1.07 +/- 0.05 m/s; the computed value was 1.130 m/s. During full closure, the leaflets showed a slight rebound, and this was also seen experimentally. For test valve B, the computed closing velocity was 1.039 m/s. In the dynamic impact analysis, the physiological pressure waveform was obtained at a normal heart rate of 70 beats/min from the mock flow test rig. Dynamic stress and displacement of the model valve were calculated as the valve was closing. The time step of calculation was determined by the wave propagation velocity and element size. With an interhinge distance of 4.966 mm based on the geometric design of the valve, maximum dynamic von Mises stress appeared near the hinge of the leaflet (26.92 MPa for valve A; 22.36 MPa for valve B). By varying the position of the hinge/pivoting axis (+/- 10%), an optimized valve geometry could be obtained based on minimal impact stress on the valve leaflet. CONCLUSION: Based on closing velocity comparison of valve A, the calculated model and loading conditions were seen to be reasonable. Computational accuracy was satisfied. The tapering feature of the leaflet is designed especially for minimal impact stress at the leaflet contact areas upon impact with the inner walls of the BMHV. These points provide an optimum structure design for the Nanyang Technological University BMHV. PMID- 12578345 TI - Kinematics of synthetic flexible leaflet heart valves during accelerated testing. AB - BACKGROUND AND AIMS OF THE STUDY: The study aim was to examine the effect of variation of test frequencies on the duration and kinematics of valve leaflets during opening, and at the fully open position. METHODS: Trileaflet polyurethane heart valves were tested in a Rowan Ash fatigue accelerator at 5, 10 and 15 Hz. Images of the valves were acquired using a high-speed camera and assessed qualitatively during opening and in the fully open position. The bending strains along the leaflet free edge and leaflet belly were also calculated from these images. Results were compared with those found at 1 Hz in a pulsatile flow simulator. The time duration of each phase of the cycle and the strain time integral when opening were also determined. RESULTS: In the fully open position, the maximum bending strain found at the commissural regions of the valve was not dependent on the cycle rate. During opening, one leaflet always deformed more than either of the others, but this leaflet was not necessarily the same one each time. The opening characteristics of the valves were dependent on cycle rate: increasing bending was observed in the belly of the leaflet with increasing frequency. The ratio of time of opening of the leaflets to time whilst fully open increased with the cycle rate, and the period of time when the valve was fully open and fully closed reduced markedly with increased cycle rate. CONCLUSION: Leaflet kinematics, bending strains, the proportion of time the valve was fully closed or open, and the strain time integral were dependent on cycle rate in accelerated testing. This may influence the relative severity of accelerated testers. PMID- 12578346 TI - Severe bioprosthetic mitral valve endocarditis due to Candida albicans. PMID- 12578347 TI - A conformational trigger for activation of a G protein by a G protein-coupled receptor. AB - G protein-coupled receptors (GPCRs) are a family of seven transmembrane helical proteins that initiate a cellular response to an environmental signal. Once activated by an extracellular signal, GPCRs trigger the intracellular signal transduction cascade by activating a heterotrimeric G protein. The interaction between the G protein and the receptor, which triggers the signal transduction, is the focus of intense interest. Three-dimensional structures of the ground state of only one GPCR, rhodopsin, are currently available, but since the G protein cannot bind to this structure, these structures did not lead to an understanding of the activation process. The recent publication of an excited state structure for the same GPCR (and comparison to the ground state structures), in conjunction with other recent biochemical data, provides new insight into G protein activation. We find that the structure data and the biochemical data, for the first time, point to a specific mode of interaction between the G protein and the receptor. Furthermore, we find that transducin (G(t)) must alter its conformation to bind to the activated receptor; the "lock and key" fit heretofore expected is likely not the correct model. We suggest that a conformational distortion, driven by the energy of binding, is induced in G(t) when it binds to the activated receptor. The conformational change in turn enables the exchange of GTP for GDP and the dissociation of the subunits. This is an example of "induced fit" originally proposed by Koshland to describe enzyme substrate interactions. PMID- 12578348 TI - A molecular motor or a regulator? Dynamin's in a class of its own. PMID- 12578349 TI - Aromatic stacking in the sugar binding site of the lactose permease. AB - Major determinants for substrate recognition by the lactose permease of Escherichia coli are at the interface between helices IV (Glu126, Ala122), V (Arg144, Cys148), and VIII (Glu269). We demonstrate here that Trp151, one turn of helix V removed from Cys148, also plays an important role in substrate binding probably by aromatic stacking with the galactopyranosyl ring. Mutants with Phe or Tyr in place of Trp151 catalyze active lactose transport with time courses nearly the same as wild type. In addition, apparent K(m) values for lactose transport in the Phe or Tyr mutants are only 6- or 3-fold higher than wild type, respectively, with a comparable V(max). Surprisingly, however, binding of high-affinity galactoside analogues is severely compromised in the mutants; the affinity of mutant Trp151-->Phe or Trp151-->Tyr is diminished by factors of at least 50 or 20, respectively. The results demonstrate that Trp151 is an important component of the binding site, probably orienting the galactopyranosyl ring so that important H-bond interactions with side chains in helices IV, V, and VIII can be realized. The results are discussed in the context of a current model for the binding site. PMID- 12578350 TI - Cholesterol dynamics in membranes of raft composition: a molecular point of view from 2H and 31P solid-state NMR. AB - Lipidic membrane systems that have been reported to be composed of sphingomyelin (SM)-cholesterol (Chol) microdomains or "rafts" by Dietrich et al. [palmitoyloleoyl-phosphatidylcholine(POPC)/SM/Chol, 1/1/1; Dietrich, C., Bagatolli, L. A., Volovyk, Z. N., Thompson, N. L., Levi, M., Jacobson, K., and Gratton, E. (2001) Biophys. J. 80, 1417-1428] and by Schroeder et al. [SCRL: Liver-PC/Liver-phosphatidylethanolamine/SM/Cerebrosides/Chol, 1/1/1/1/2; Schroeder, R., London, E., and Brown, D. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 12130-12134] were investigated under the form of fully hydrated liposomes by the noninvasive solid-state (31)P and (2)H NMR method. Liposomes of binary lipid composition POPC/Chol and SM/Chol were also studied as boundary/control systems. All systems are found to be in the liquid-ordered phase (Lo) at physiological temperatures. Use of deuterium-labeled cholesterol afforded finding both the position of the sterol motional axis and its molecular order parameter. The axis of anisotropic rotation of cholesterol is such that the molecule is, on average, quasiperpendicular to the membrane plane, in all of the four systems investigated. Cholesterol order parameters greater than 0.8 are observed, indicating that the sterol is in a very motionally restricted environment in the temperature range 0-60 degrees C. The binary mixtures present "boundary" situations with the lowest values for POPC/Chol and the highest for SM/Chol. The SCRL raft mixture has the same ordering as the SM/Chol, i.e., the highest order parameter values over the temperature range. It demonstrates that in the SCRL mixture cholesterol dynamics is as in the binary system SM/Chol, therefore, suggesting that it might be depleted from the rest of the membrane to form complexes as if it were alone with SM. On the other hand, the mixture POPC/SM/Chol exhibits an intermediate ordering situation between those of SM/Chol and POPC/Chol. This strongly suggests that cholesterol could be in fast exchange, at the NMR time scale (milli- to microseconds), between two or more membrane regions of different dynamics and questions the statement of "rigid domains" made of SM and cholesterol in the model "raft" system POPC/SM/Chol. PMID- 12578351 TI - Substrate-induced conformational changes of the periplasmic N-terminus of an outer-membrane transporter by site-directed spin labeling. AB - The structure and dynamics of the N-terminal and core regions of BtuB, an outer membrane vitamin B(12) transporter from Escherichia coli, were investigated by site-directed spin labeling. Cysteine mutants were generated by site-directed mutagenesis to place spin labels in the N-terminal region (residues 1-17), the core region (residues 25-30), and double labels into the Ton box (residues 6-12). BtuB mutants were expressed, spin labeled, purified, and reconstituted into phosphatidylcholine. In the presence of substrate (vitamin B(12)), EPR spectroscopy demonstrates that there is a conformational change in the Ton box similar to that seen previously for BtuB in intact outer membranes. The Ton box is positioned within the beta-barrel of BtuB in the absence of substrate (docked configuration) but becomes unfolded and increases its aqueous exposure upon substrate binding (undocked configuration). This conformational change and the similarity in the EPR spectra between reconstituted and native membranes indicate that BtuB is correctly folded and functional in the reconstituted system. The protein segment on the N-terminal side of the Ton box is highly mobile, and it becomes more mobile in the presence of substrate. Side chains in the region C terminal to the Ton box also show increases in mobility with substrate addition, but position 16 appears to define a hinge point for this conformation change. EPR line shapes and relaxation data indicate that residues 25-30 form a beta-strand structure, which is analogous to the first beta-strand in the cores of the homologous iron transporters. When substrate binds to BtuB, this first beta strand remains folded. The EPR spectra of double-nitroxide labels within the Ton box are broadened because of dipolar and collisional exchange interactions. The broadening pattern indicates that the Ton box is not helical but is in an extended or beta-strand structure. PMID- 12578352 TI - The structure, cation binding, transport, and conductance of Gly15-gramicidin A incorporated into SDS micelles and PC/PG vesicles. AB - To further investigate the effect of single amino acid substitution on the structure and function of the gramicidin channel, an analogue of gramicidin A (GA) has been synthesized in which Trp(15) is replaced by Gly in the critical aqueous interface and cation binding region. The structure of Gly(15)-GA incorporated into SDS micelles has been determined using a combination of 2D-NMR spectroscopy and molecular modeling. Like the parent GA, Gly(15)-GA forms a dimeric channel composed of two single-stranded, right-handed beta(6.3)-helices joined by hydrogen bonds between their N-termini. The replacement of Trp(15) by Gly does not have a significant effect on backbone structure or side chain conformations with the exception of Trp(11) in which the indole ring is rotated away from the channel axis. Measurement of the equilibrium binding constants and Delta G for the binding of monovalent cations to GA and Gly(15)-GA channels incorporated into PC vesicles using (205)Tl NMR spectroscopy shows that monovalent cations bind much more weakly to the Gly(15)-GA channel entrance than to GA channels. Utilizing the magnetization inversion transfer NMR technique, the transport of Na(+) ions through GA and Gly(15)-GA channels incorporated into PC/PG vesicles has been investigated. The Gly(15) substitution produces an increase in the activation enthalpy of transport and thus a significant decrease in the transport rate of the Na(+) ion is observed. The single-channel appearances show that the conducting channels have a single, well-defined structure. Consistent with the NMR results, the single-channel conductances are reduced by 30% and the lifetimes by 70%. It is concluded that the decrease in cation binding, transport, and conductance in Gly(15)-GA results from the removal of the Trp(15) dipole and, to a lesser extent, the change in orientation of Trp(11). PMID- 12578353 TI - Solution structure of a cis-opened (10R)-N6-deoxyadenosine adduct of (9S,10R) 9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene in a DNA duplex. AB - The solution structure of an 11-mer DNA duplex, d(CGGTCA*CGAGG) x d(CCTCGTGACCG), containing a 10R adduct at dA* that corresponds to the cis addition of the N(6) amino group of dA(6) to (+)-(9S,10R)-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene was studied by 2D NMR methods. The NOESY cross-peak patterns indicate that the hydrocarbon is intercalated on the 5'-side of the modified base. This observation is the same as that observed for other oligonucleotides containing (10R)-dA adducts but opposite to that observed for the corresponding (10S)-dA adducts which are intercalated on the 3'-side of the modified base. The hydrocarbon is intercalated from the major groove without significant disruption of either the anti glycosidic torsion angle of the modified residue or the base pairing of the modified residue with the complementary residue on the opposite strand. The ensemble of 10 structures determined exhibits relatively small variations (6-15 degrees) in the characteristic hydrocarbon-base dihedral angles (alpha' and beta') as well as the glycosidic torsion angle chi. These angles are similar to those in a previously determined cis-opened benzo[a]pyrene diol epoxide-(10R)-dA adduct structure. Comparison of the present structure with the cis-opened diol epoxide adduct suggests that the absence of the 7- and 8-hydroxyl groups results in more efficient stacking of the aromatic moiety with the flanking base pairs and deeper insertion of the hydrocarbon into the helix. Relative to normal B-DNA, the duplex containing the present tetrahydroepoxide adduct is unwound at the lesion site, whereas the diol epoxide adduct structure is more tightly wound than normal B-DNA. Buckling of the adducted base pair as well as the C(5)-G(18) base pair that lies immediately above the hydrocarbon is much less severe in the present adducted structure than its cis-opened diol epoxide counterpart. PMID- 12578354 TI - Investigations of Sso7d catalytic residues by NMR titration shifts and electrostatic calculations. AB - Sso7d is a small basic protein consisting of 62 amino acids isolated from the thermoacidophilic archeobacterium Sulfolobus solfataricus. The protein is endowed with DNA binding properties, RNase activity, and the capability of rescuing aggregated proteins in the presence of ATP. In this study, the electrostatic properties of Sso7d are investigated by using the Poisson-Boltzmann calculation of the surface potential distribution and following by NMR spectroscopy the proton chemical shift pH titration of acidic residues. Although the details of the catalytic mechanism still have to be defined, the results from NMR experiments confirm the possible involvement of Glu35 as the proton acceptor in the catalytic reaction, as seen by its abnormally high pK(a) value. Poisson Boltzmann calculations and NMR titration shifts suggest the presence of a possible hydrogen bond between Glu35 and Tyr33, with a consequent rather rigid arrangement at these positions. Comparison with RNase T1 suggests that Tyr7 may be a good candidate for acting as a proton donor in the active site of Sso7d as shown by its low phenolic pK(a) of approximately 9.3. Titration experiments performed with the UpA, a RNA dinucleotide model, showed that the protein residues affected by the interaction are mainly located in a different region with respect to the surface affected by DNA recognition, in good agreement with the surface potential distribution found with electrostatic calculations. PMID- 12578355 TI - 2.1 A crystal structure of human PXR in complex with the St. John's wort compound hyperforin. AB - The nuclear xenobiotic receptor PXR is activated by a wide variety of clinically used drugs and serves as a master regulator of drug metabolism and excretion gene expression in mammals. St. John's wort is used widely in Europe and the United States to treat depression. This unregulated herbal remedy leads to dangerous drug-drug interactions, however, in patients taking oral contraceptives, antivirals, or immunosuppressants. Such interactions are caused by the activation of the human PXR by hyperforin, the psychoactive agent in St. John's wort. In this study, we show that hyperforin induces the expression of numerous drug metabolism and excretion genes in primary human hepatocytes. We present the 2.1 A crystal structure of hyperforin in complex with the ligand binding domain of human PXR. Hyperforin induces conformational changes in PXR's ligand binding pocket relative to structures of human PXR elucidated previously and increases the size of the pocket by 250 A(3). We find that the mutation of individual aromatic residues within the ligand binding cavity changes PXR's response to particular ligands. Taken together, these results demonstrate that PXR employs structural flexibility to expand the chemical space it samples and that the mutation of specific residues within the ligand binding pocket of PXR tunes the receptor's response to ligands. PMID- 12578356 TI - C-terminal half of human centrin 2 behaves like a regulatory EF-hand domain. AB - Human centrin 2 (HsCen2) is an EF-hand protein that plays a critical role in the centrosome duplication and separation during cell division. We studied the structural and Ca(2+)-binding properties of two C-terminal fragments of this protein: SC-HsCen2 (T94-Y172), covering two EF-hands, and LC-HsCen2 (M84-Y172), having 10 additional residues. Both fragments are highly disordered in the apo state but become better structured (although not conformationally homogeneous) in the presence of Ca(2+) and depending on the nature of the cations (K(+) or Na(+)) in the buffer. Only the longer C-terminal domain, in the Ca(2+)-saturated state and in the presence of Na(+) ions, was amenable to structure determination by nuclear magnetic resonance. The solution structure of LC-HsCen2 reveals an open two EF-hand structure, similar to the conformation of related Ca(2+)-saturated regulatory domains. Unexpectedly, the N-terminal helix segment (F86-T94) lies over the exposed hydrophobic cavity. This unusual intramolecular interaction increases considerably the Ca(2+) affinity and constitutes a useful model for the target binding. PMID- 12578357 TI - Mutational analysis of the complex of human RNase inhibitor and human eosinophil derived neurotoxin (RNase 2). AB - RNase inhibitor (RI) binds diverse proteins in the pancreatic RNase superfamily with extremely high avidity. Previous studies showed that tight binding of RNase A and angiogenin (Ang) is achieved primarily through interactions of hot spot residues in the 434-460 C-terminal segment of RI with the enzymatic active site; Asp435 of RI forms key hydrogen bonds with the catalytic lysine in both complexes, whereas the other contacts are largely distinctive. Here we have investigated the structural basis for recognition of a third ligand, eosinophil derived neurotoxin (EDN), by single-site and multisite mutagenesis. Surprisingly, Ala replacement of Asp435 decreases affinity for EDN only by 14-fold, as compared to the several hundred-fold decreases with RNase A and Ang, and individual mutations of three other hot spot residues-Tyr434, Tyr437, and Ser460-have essentially no effect. Ala substitutions of nine additional residues, selected by examining a computational model of the RI.EDN complex, also have no marked impact. Overall, the losses in affinity for the single-residue variants examined account for only approximately 25% of the free energy of binding for the complex. However, multisite mutagenesis of RI reveals strong superadditivity of mutational effects, indicating that part of this shortfall reflects negative cooperativity. Replacement of Tyr434 together with Asp435 or Tyr437 increases K(i) by 540- and 290-fold, respectively. Thus, the C-terminal region of RI again plays an important role in ligand recognition, although probably smaller than for binding RNase A and Ang. Simultaneous substitutions of three neighboring tryptophans (261, 263, and 318) on RI attenuate affinity even more dramatically (by 4900 fold), indicating that the interactions of this RI region also contribute a considerable amount of the binding energy for the EDN complex. These findings highlight the potential importance of cooperativity in protein-protein interactions and the consequent limitations of single-site mutagenesis for assessing interface energetics. PMID- 12578358 TI - Solution structure of the carboxyl-terminal domain of RAP74 and NMR characterization of the FCP1-binding sites of RAP74 and human TFIIB. AB - FCP1 (TFIIF-associated CTD phosphatase) is the only known phosphatase specific for the phosphorylated CTD of RNAP II. The phosphatase activity of FCP1 is strongly enhanced by the carboxyl-terminal domain of RAP74 (cterRAP74, residues 436-517), and this stimulatory effect of TFIIF can be blocked by TFIIB. It has been shown that cterRAP74 and the core domain of hTFIIB (TFIIBc, residues 112 316) directly interact with the carboxyl-terminal domain of hFCP1 (cterFCP, residues 879-961), and these interactions may be responsible for the regulatory activities of TFIIF and TFIIB on FCP1. We have determined the NMR solution structure of human cterRAP74, and we have used NMR methods to map the cterFCP binding sites for both cterRAP74 and human TFIIB. We show that cterFCP binds to a groove of cterRAP74 between alpha-helices H2 and H3, without affecting the secondary structure of cterRAP74. We also show that cterFCP binds to a groove of TFIIBc between alpha-helices D1 and E1 in the first cyclin repeat. We find that the cterFCP-binding site of TFIIBc is very similar to the binding site for the HSV transcriptional activator protein VP16 on the first cyclin repeat of TFIIBc. The cterFCP-binding sites of both RAP74 and TFIIBc form shallow grooves on the protein surface, and they are both rich in hydrophobic and positively charged amino acid residues. These results provide new information about the recognition of acidic-rich activation domains involved in transcriptional regulation, and provide insights into how TFIIF and TFIIB regulate the FCP1 phosphatase activity in vivo. PMID- 12578360 TI - Kinetics and energetics of the binding between barley alpha-amylase/subtilisin inhibitor and barley alpha-amylase 2 analyzed by surface plasmon resonance and isothermal titration calorimetry. AB - The kinetics and energetics of the binding between barley alpha amylase/subtilisin inhibitor (BASI) or BASI mutants and barley alpha-amylase 2 (AMY2) were determined using surface plasmon resonance and isothermal titration calorimetry (ITC). Binding kinetics were in accordance with a 1:1 binding model. At pH 5.5, [Ca(2+)] = 5 mM, and 25 degrees C, the k(on) and k(off) values were 8.3 x 10(+4) M(-1) s(-1) and 26.0 x 10(-4) s(-1), respectively, corresponding to a K(D) of 31 nM. K(D) was dependent on pH, and while k(off) decreased 16-fold upon increasing pH from 5.5 to 8.0, k(on) was barely affected. The crystal structure of AMY2-BASI shows a fully hydrated Ca(2+) at the protein interface, and at pH 6.5 increase of [Ca(2+)] in the 2 microM to 5 mM range raised the affinity 30-fold mainly due to reduced k(off). The K(D) was weakly temperature dependent in the interval from 5 to 35 degrees C as k(on) and k(off) were only increasing 4- and 12-fold, respectively. A small salt dependence of k(on) and k(off) suggested a minor role for global electrostatic forces in the binding and dissociation steps. Substitution of a positively charged side chain in the mutant K140L within the AMY2 inhibitory site of BASI accordingly did not change k(on), whereas k(off) increased 13-fold. ITC showed that the formation of the AMY2-BASI complex is characterized by a large exothermic heat (Delta H = -69 +/- 7 kJ mol( 1)), a K(D) of 25 nM (27 degrees C, pH 5.5), and an unfavorable change in entropy (-T Delta S = 26 +/- 7 kJ mol(-1)). Calculations based on the thermodynamic data indicated minimal structural changes during complex formation. PMID- 12578359 TI - Structural basis for a lethal mutation in U6 RNA. AB - U6 RNA is essential for nuclear pre-mRNA splicing and has been implicated directly in catalysis of intron removal. The U80G mutation at the essential magnesium binding site of the U6 3' intramolecular stem-loop region (ISL) is lethal in yeast. To further understand the structure and function of the U6 ISL, we have investigated the structural basis for the lethal U80G mutation by NMR and optical spectroscopy. The NMR structure reveals that the U80G mutation causes a structural rearrangement within the ISL resulting in the formation of a new Watson-Crick base pair (C67 x G80), and disrupts a protonated C67 x A79 wobble pair that forms in the wild-type structure. Despite the structural change, the accessibility of the metal binding site is unperturbed, and cadmium titration produces similar phosphorus chemical shift changes for both the U80G mutant and wild-type RNAs. The thermodynamic stability of the U80G mutant is significantly increased (Delta Delta G(fold) = -3.6 +/- 1.9 kcal/mol), consistent with formation of the Watson-Crick pair. Our structural and thermodynamic data, in combination with previous genetic data, suggest that the lethal basis for the U80G mutation is stem-loop hyperstabilization. This hyperstabilization may prevent the U6 ISL melting and rearrangement necessary for association with U4. PMID- 12578361 TI - Intramolecular proton-transfer reactions in a membrane-bound proton pump: the effect of pH on the peroxy to ferryl transition in cytochrome c oxidase. AB - In the membrane-bound redox-driven proton pump cytochrome c oxidase, electron- and proton-transfer reactions must be coupled, which requires controlled modulation of the kinetic and/or thermodynamic properties of proton-transfer reactions through the membrane-spanning part of the protein. In this study we have investigated proton-transfer reactions through a pathway that is used for the transfer of both substrate and pumped protons in cytochrome c oxidase from Rhodobacter sphaeroides. Specifically, we focus on the formation of the so-called F intermediate, which is rate limited by an internal proton-transfer reaction from a possible branching point in the pathway, at a glutamic-acid residue (E(I 286)), to the binuclear center. We have also studied the reprotonation of E(I 286) from the bulk solution. Evaluation of the data in terms of a model presented in this work gives a rate of internal proton transfer from E(I-286) to the proton acceptor at the catalytic site of 1.1 x 10(4) s(-1). The apparent pK(a) of the donor (E(I-286)), determined from the pH dependence of the F-formation kinetics, was found to be 9.4, while the pK(a) of the proton acceptor at the catalytic site is likely to be > or = 2.5 pH units higher. In the pH range up to pH 10 the proton equilibrium between the bulk solution and E(I-286) was much faster than 10(4) s(-1), while in the pH range above pH 10 the proton uptake from solution is rate limiting for the overall reaction. The apparent second-order rate constant for proton transfer from the bulk solution to E(I-286) is >10(13) M(-1) s(-1), which indicates that the proton uptake is assisted by a local buffer consisting of protonatable residues at the protein surface. PMID- 12578362 TI - Protein-protein interactions between cytochrome b and the Fe-S protein subunits during QH2 oxidation and large-scale domain movement in the bc1 complex. AB - The ubihydroquinone:cytochrome (cyt) c oxidoreductase, or bc(1) complex, and its homologue the b(6)f complex are key components of respiratory and photosynthetic electron transport chains as they contribute to the generation of an electrochemical gradient used by the ATP synthase to produce ATP. The bc(1) complex has two catalytic domains, ubihydroquinone oxidation (Q(o)) and ubiquinone reduction (Q(i)) sites, that are located on each side of the membrane. The key to the energetic efficiency of this enzyme relies upon the occurrence of a unique electron bifurcation reaction at its Q(o) site. Recently, several lines of evidence have converged to establish that in the bc(1) complex the extrinsic domain of the Fe-S subunit that contains a [2Fe2S] metal cluster moves during catalysis to shuttle electrons between the Q(o) site and c(1) heme. While this step is required for electron bifurcation, available data also suggest that the movement might be controlled to ensure maximal energetic efficiency [Darrouzet et al. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 4567-4572]. To gain insight into the plausible control mechanism, we used a biochemical genetic approach to define the different regions of the bc(1) complex that might interact with each other. Previously, we found that a mutation located at position L286 of the ef loop of Rhodobacter capsulatus cyt b could alleviate movement impairment resulting from a mutation in the hinge region, linking the [2Fe2S] cluster domain to the membrane anchor of the Fe-S subunit. Here we report that various substitutions at position 288 on the opposite side of the ef loop also impair Q(o) site catalysis. In particular, we note that while most of the substitutions affect only QH(2) oxidation, yet others like T288S also hinder the rate of the movement of the Fe-S subunit. Thus, position 288 of cyt b appears to be important for both the QH(2) oxidation and the movement of the Fe-S subunit. Moreover, we found that, upon substitution of T288 by other amino acids, additional compensatory mutations located at the [2Fe2S] cluster or the hinge domains of the Fe-S subunit, or on the cd loop of cyt b, arise readily to alleviate these defects. These studies indicate that intimate protein-protein interactions occur between cyt b and the Fe-S subunits to sustain fast movement and efficient QH(2) oxidation and highlight the critical dual role the ef loop of cyt b to fine-tune the docking and movement of the Fe-S subunit during Q(o) site catalysis. PMID- 12578363 TI - Altered Cu metabolism and differential transcription of Cu/ZnSod genes in a Cu/ZnSOD-deficient mutant of maize: evidence for a Cu-responsive transcription factor. AB - Maize inbred line A351 exhibits extremely low levels of Cu/Zn superoxide dismutase (SOD) isozymes, three cytosolic and one chloroplastic, which are increased by supplying copper to near-toxic concentrations. Activities of the copper enzymes cytochrome c oxidase and ascorbate oxidase are also reduced. The level of expression of the maize copper chaperone for SOD is normal to elevated. The gene transcript encoding chloroplastic SOD-1 is present at normal levels, whereas RNA levels of the cytosolic SODs are low and increase with added copper, suggesting a promoter element and copper-dependent transcription factor common to the three genes. Although a reduced level of high-affinity copper transport in A351 cannot be ruled out, high transcript levels of a constitutively expressed metallothionein, suggesting increased copper chelation capacity and creating a general copper-deprivation effect, seem to be a likely cause of the reduced levels of copper enzyme activity and Cu/ZnSod gene transcripts. While exogenous copper does not affect the wild-type SOD activity or protein, it increases wild type Cu/ZnSod transcript levels in a response similar to that of several yeast genes involved in copper sequestration and antioxidant defense. A sequence that is highly homologous to those of the copper-responsive transcription factors ACE1 (Saccharomyces cerevisiae) and AMT1 (Candida glabrata) is present in the promoters of three maize Cu/ZnSod genes. PMID- 12578364 TI - Structure-based improvement of the biophysical properties of immunoglobulin VH domains with a generalizable approach. AB - In a systematic study of V gene families carried out with consensus V(H) and V(L) domains alone and in combinations in the scFv format, we found comparatively low expression yields and lower cooperativity in equilibrium unfolding in antibody fragments containing V(H) domains of human germline families 2, 4, and 6. From an analysis of the packing of the hydrophobic core, the completeness of charge clusters, the occurrence of unsatisfied hydrogen bonds, and residues with low beta-sheet propensities, positive Phi angles, and exposed hydrophobic side chains, we pinpointed residues potentially responsible for the unsatisfactory properties of these germline-encoded sequences. Several of those are in common between the domains of the even-numbered subgroups, but do not occur in the odd numbered ones. In this study, we have systematically exchanged those residues alone and in combination in two different scFvs using the V(H)6 framework, and we describe their effect on equilibrium stability and folding yield. We improved the stability by 20.9 kJ/mol and the expression yield by a factor of 4 and can now use these data to rationally engineer antibodies derived from this and similar germline families for better biophysical properties. Furthermore, we provide an improved design for libraries exploiting the significant additional diversity provided by these frameworks. Both antibodies studied here completely retain their binding affinity, demonstrating that the CDR conformations were not affected. PMID- 12578365 TI - Inhibition of beta-lactamases by monocyclic acyl phosph(on)ates. AB - The cyclic acyl phosph(on)ates, 1-hydroxy-5-phenyl-2,6-dioxaphosphorinone(3)-1 oxide, its 4-phenyl isomer, and the phosphonate (2-oxo) analogue of the latter inhibited typical class A (TEM-2) and class C (Enterobacter cloacae P99) beta lactamases in a time-dependent fashion. No enzyme-catalyzed turnover was detected in any case. The interactions occurring were interpreted in terms of the reaction scheme E + I left arrow over right arrow EI left arrow over right arrow EI', where EI is a reversibly formed noncovalent complex, and EI' is a covalent complex. Reactions of the cyclic phosphates with the P99 beta-lactamase were effectively irreversible, while that of the 4-phenyl cyclic phosphate with the TEM beta-lactamase was slowly reversible. The 4-phenyl cyclic phosphate was generally the most effective inhibitor, both kinetically and thermodynamically, with second-order rate constants of inactivation of both enzymes around 10(4) s( 1) M(-1). This compound also bound noncovalently to both enzymes, with dissociation constants of 25 microM from the P99 enzyme and 100 microM from the TEM. It is unusual to find an inhibitor equally effective against the TEM and P99 enzymes; the beta-lactamase inhibitors currently employed in medical practice (e.g., clavulanic acid) are significantly more effective against class A enzymes. The results of lysinoalanine analysis after hydroxide treatment of the inhibited enzymes and of a (31)P nuclear magnetic resonance spectrum of one such complex were interpreted as favoring a mechanism of inactivation by enzyme acylation rather than phosphylation. Molecular modeling of the enzyme complexes of the 4 phenyl phosphate revealed bound conformations where recyclization and thus reactivation of the enzyme would be difficult. The compounds studied were turned over slowly or not at all by acetylcholinesterase and phosphodiesterase I. PMID- 12578366 TI - Binding of ATP as well as tetrahydrofolate induces conformational changes in Lactobacillus casei folylpolyglutamate synthetase in solution. AB - Folylpolyglutamate synthetase (FPGS) catalyzes the addition of glutamate to folate derivatives to form folate polyglutamates. FPGS is essential for folate biosynthesis in bacteria and retention of folate pools in eukaryotes. X-ray crystallographic analyses of binary and ternary complexes of Lactobacillus casei FPGS suggest that binding of folate triggers a conformational change that activates FPGS. We used EPR and CD spectroscopy to further characterize the conformational change in the FPGS reaction. For EPR spectroscopy, two cysteine residues were introduced into FPGS by site-directed mutagenesis, K172C in the N terminal domain and D345C in the C-terminal domain. The mutant protein was expressed, purified, and labeled with methanethiosulfonate. Addition of ATP, tetrahydrofolate, or 5,10-methylenetetrahydrofolate but not glutamate to FPGS showed broadening of EPR spectra, which is due to stronger spin-spin interactions, suggesting that both ATP and tetrahydrofolates cause a conformational change. ATP binding had an EPR spectrum distinct from that of tetrahydrofolate binding, indicating that it caused a different conformational change. When both ATP and THF were bound, the spectrum was identical to that seen when THF alone bound to the enzyme, showing that the THF-induced conformation was dominant. The spectral broadening suggests that the conformation change involves the two domains moving closer together, which is consistent with the rigid-body rotation of the C-terminal domain observed in the FPGS crystal structure with AMPPCP and 5,10-methylenetetrahydrofolate bound. No changes in the CD spectra were observed with the addition of FPGS substrates, suggesting that the conformational changes did not affect the secondary structure elements of the enzyme. These studies confirm the conformational change seen in the crystal structure by an independent method but also show that ATP binds to the free enzyme and affects its conformation. PMID- 12578367 TI - Binding of 2-aryl-4-(piperidin-1-yl)butanamines and 1,3,4-trisubstituted pyrrolidines to human CCR5: a molecular modeling-guided mutagenesis study of the binding pocket. AB - The results of investigations in these laboratories of 2-aryl-4-(piperidin-1 yl)butanamines and 1,3,4-trisubstituted pyrrolidines as human CCR5 antagonists have recently been disclosed. To facilitate further development of these antagonists, we have developed a pharmacophore model based on the structure activity relationships (SAR) and a human CCR5 receptor docking model using the crystal structure of rhodopsin as a template [Palczewski, K., et al. (2000) Science 289, 739-745]. Guided by the receptor docking model, we have mapped the compounds' site of interaction with CCR5 using site-directed mutagenesis experiments. Our results are consistent with a binding site for the two series that is located within a cavity near the extracellular surface formed by transmembrane helices 2, 3, 6, and 7. This site is overlapping yet distinct from that reported for another antiviral agent which binds to CCR5 [Dragic, T., et al. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 5639-5644]. PMID- 12578368 TI - Surface expansion is independent of and occurs faster than core solvation during the unfolding of barstar. AB - The denaturant-induced unfolding kinetics of the 89-residue protein, barstar, have been examined using fluorescence resonance energy transfer (FRET) at 25 degrees C and pH 8.0. The core tryptophan, Trp53, in barstar serves as a fluorescence donor, and a thionitrobenzoic acid moiety (TNB) attached to a cysteine residue acts as an acceptor to form an efficient FRET pair. Four different single-cysteine containing mutants of barstar with cysteine residues at positions 25, 40, 62, and 82 were studied. The unfolding kinetics of the four mutant forms of barstar were monitored by measurement of the changes in the fluorescence intensity of Trp53 in the unlabeled and TNB-labeled proteins. The rate of change of fluorescence of the single-tryptophan residue, Trp53, in the unlabeled protein, where no FRET occurs, yields the rate of solvation of the core. This rate is similar for all four unlabeled proteins. The rate of the increase in the fluorescence of Trp53 in the labeled protein, where FRET from the tryptophan to the TNB label occurs, yields the rate of decrease in FRET efficiency during unfolding. The decrease in FRET efficiency for proteins labeled at either of the two buried positions (Cys40 or Cys82) occurs at a rate similar to the rate of core solvation. The decrease in FRET efficiency for the acceptor at Cys40 is also shown to be sensitive to the isomerization of the Tyr47-Pro48 cis bond. For the proteins where the label is at a solvent-exposed position (Cys25 and Cys62), the decrease in FRET efficiency occurs in two kinetic phases; 15-25% of the FRET efficiency decreases in the faster phase, and the remaining FRET efficiency decreases in a slower phase, the rate of which is the same as the rate of core solvation. These results clearly indicate that, during unfolding, the protein surface expands faster than, and independently of, water intrusion into the core. PMID- 12578369 TI - Structural and biochemical exploration of a critical amino acid in human 8 oxoguanine glycosylase. AB - Members of the HhH-GPD superfamily of DNA glycosylases are responsible for the recognition and removal of damaged nucleobases from DNA. The hallmark of these proteins is a motif comprising a helix-hairpin-helix followed by a Gly/Pro-rich loop and terminating in an invariant, catalytically essential aspartic acid residue. In this study, we have probed the role of this Asp in human 8-oxoguanine DNA glycosylase (hOgg1) by mutating it to Asn (D268N), Glu (D268E), and Gln (D268Q). We show that this aspartate plays a dual role, acting both as an N terminal alpha-helix cap and as a critical residue for catalysis of both base excision and DNA strand cleavage by hOgg1. Mutation of this residue to asparagine, another helix-capping residue, preserves stability of the protein while drastically reducing enzymatic activity. A crystal structure of this mutant is the first to reveal the active site nucleophile Lys249 in the presence of lesion-containing DNA; this structure offers a tantalizing suggestion that base excision may occur by cleavage of the glycosidic bond and then attachment of Lys249. Mutation of the aspartic acid to glutamine and glutamic acid destabilizes the protein fold to a significant extent but, surprisingly, preserves catalytic activity. Crystal structures of these mutants complexed with an unreactive abasic site in DNA reveal these residues to adopt a sterically disfavored helix-capping conformation. PMID- 12578370 TI - Dynamic roles of arginine residues 82 and 92 of Escherichia coli 6-hydroxymethyl 7,8-dihydropterin pyrophosphokinase: crystallographic studies. AB - 6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) catalyzes the pyrophosphoryl transfer from ATP to 6-hydroxymethyl-7,8-dihydropterin (HP), the first reaction in the folate biosynthetic pathway. Arginine residues 82 and 92, strictly conserved in 35 HPPK sequences, play dynamic roles in the catalytic cycle of the enzyme. At 0.89-A resolution, two distinct conformations are observed for each of the two residues in the crystal structure of the wild-type HPPK in complex with two HP variants, two Mg(2+) ions, and an ATP analogue. Structural information suggests that R92 first binds to the alpha-phosphate group of ATP and then shifts to interact with the beta-phosphate as R82, which initially does not bind to ATP, moves in and binds to alpha-phosphate when the pyrophosphoryl transfer is about to occur. The dynamic roles of R82 and R92 are further elucidated by five more crystal structures of two mutant proteins, R82A and R92A, with and without bound ligands. Two oxidized forms of HP are observed with an occupancy ratio of 0.50:0.50 in the 0.89-A structure. The oxidation of HP has significant impact on its binding to the protein as well as the conformation of nearby residue W89. PMID- 12578371 TI - Catalytic roles of arginine residues 82 and 92 of Escherichia coli 6 hydroxymethyl-7,8-dihydropterin pyrophosphokinase: site-directed mutagenesis and biochemical studies. AB - The roles of a pair of conserved positively charged residues R82 and R92 at a catalytic loop of Escherichia coli 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) have been investigated by site-directed mutagenesis and biochemical analysis. In the structure of HPPK in complex with ATP and a 6 hydroxymethyl-7,8-dihydropterin (HP) analogue, the guanidinium group of R82 forms two hydrogen bonds with the alpha-phosphate and that of R92 two hydrogen bonds with the beta-phosphate. In the structure of HPPK in complex with alpha,beta methyleneadenosine triphosphate (AMPCPP, an ATP analogue) and HP, the guanidinium group of R82 has no direct interaction with AMPCPP and that of R92 forms two hydrogen bonds with the alpha-phosphate. Substitution of R82 with alanine caused a decrease in the rate constant for the chemical step by a factor of approximately 380, but there were no significant changes in the binding energy or binding kinetics of either substrate. Substitution of R92 with alanine caused a decrease in the rate constant for the chemical step by a factor of approximately 3.5 x 10(4). The mutation caused no significant changes in the binding energy or binding kinetics of MgATP. It did not cause a significant change in the binding energy of HP either but caused a decrease in the association rate constant for the binding of HP by a factor of approximately 4.5 and a decrease in the dissociation rate constant by a factor of approximately 10. The overall structures of the ternary complexes of both mutants were very similar to the corresponding structure of wild-type HPPK as described in the companion paper. The results suggest that R82 does not contribute to the binding of either substrate, and R92 is dispensable for the binding of MgATP but plays a role in facilitating the binding of HP. Both R82 and R92 are important for catalysis, and R92 plays a critical role in the transition state stabilization. PMID- 12578372 TI - The carboxyl side chain of glutamate 681 interacts with a chloride binding modifier site that allosterically modulates the dimeric conformational state of band 3 (AE1). Implications for the mechanism of anion/proton cotransport. AB - Glutamate 681 is thought to be located within the transport channel of band 3 (AE1, the chloride/bicarbonate exchanger), where it acts as a proton donor for the anion/proton cotransport function. Here we show that neutralization of the negative charge on glutamate 681 by chemically modifying band 3 with Woodward's reagent K plus sodium borohydride (i.e., the modification process) exposes a cryptic, conformationally active chloride-binding site which functions to modulate allosterically the conformational state of the band 3 dimer. Chloride binding was determined by measuring the effect of increasing chloride concentration on the rate of DBDS (4,4'-dibenzamido-2,2'-stilbenedisulfonate) release from band 3 using a stopped-flow fluorescence kinetic inhibitor replacement assay with DIDS (4,4'-diisothiocyanato-2,2'-stilbenedisulfonate) as the replacing inhibitor. The time course for DBDS release from unmodified, control band 3 was monophasic and exponential. Chloride binding to the transport site accelerated the rate of DBDS release, with the observed rate constant showing a hyperbolic dependence on chloride concentration, while the total change in reaction fluorescence remained constant. After modification of glutamate 681, DBDS release was monophasic in the absence of chloride, but the rapid addition of chloride at constant ionic strength induced a doubling in the fluorescence quantum yield for the bound DBDS molecules. This was associated with the development of 50:50 biphasic kinetics for DBDS release. Such changes were independent of the degree of modification of the band 3 subunit population between the 66% and 91% levels. Titration of the increase in total reaction fluorescence gave an apparent chloride binding K(d) of between 7 and 10 mM, which is 25-40-fold higher in affinity than chloride binding to the transport site. The dependence of the kinetic constants for both phases of the DBDS release reaction on chloride concentration was nonhyperbolic, which contrasts with unmodified band 3, and is indicative of the presence of two classes of chloride-binding sites on the modified transporter. We have also found that the fraction of subunits capable of binding DBDS reversibly, or DIDS covalently, decreased nonlinearly in the absence of chloride as the level of modification of the band 3 subunit population increased. In contrast, the same DBDS binding correlation plot showed a maximum in the presence of saturating chloride. The observation of such nonlinear correlation plots is consistent with a noncooperative dimer model for the modification process, where each dimeric species must possess different properties with respect to stilbenedisulfonate binding capacity and with respect to the spectral-kinetic response of bound stilbenedisulfonate molecules to the addition of chloride. Within the context of this model, the fractions of the three molecular dimeric species (i.e., the unmodified dimer, the dimer with one subunit modified, and the fully modified band 3 dimer) are calculated as a function of the level of modification of the band 3 subunit population. Nonlinear correlation plots are generated by then assigning the following specific properties to each dimeric species. The unmodified dimer binds DBDS but does not change its fluorescence quantum yield upon addition of chloride. The half modified dimer binds DBDS on both modified and unmodified subunits, and both of those DBDS molecules increase their fluorescence quantum yield by 2-fold when chloride is added, and the system develops 50:50 biphasic DBDS release kinetics. Finally, the model requires that the fully modified dimer does not bind DBDS or DIDS. This model generates theoretical correlation plots that can represent the data presented in this study. We propose that neutralization of glutamate 681 on the half-modified band 3 dimer exposes an allosteric, chloride-binding modifier site which functions to facilitate the anion/proton cotransport process (a) by blocking the "redocking" of the carboxyl side chain of glutamate (thus raising its pK) and (b) by inducing amate (thus raising its pK) and (b) by inducing a conformational change in the band 3 dimer from a symmetrical to an asymmetrical state. PMID- 12578373 TI - Altering substrate specificity of phosphatidylcholine-preferring phospholipase C of Bacillus cereus by random mutagenesis of the headgroup binding site. AB - PLC(Bc) is a 28.5 kDa monomeric enzyme that catalyzes the hydrolysis of the phosphodiester bond of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine to provide a diacylglycerol and the corresponding phosphorylated headgroup. Because single replacements of Glu4, Tyr56, and Phe66 in the headgroup binding pocket led to changes in substrate specificity [Martin et al. (2000) Biochemistry 39, 3410-3415], a combinatorial library of approximately 6000 maltose binding protein-PLC(Bc) fusion protein mutants containing random permutations of these three residues was generated to identify PLC(Bc) mutants with altered specificity profiles and high catalytic activities. Members of this library were screened for hydrolytic activity toward the water soluble substrates C6PC, C6PE, and C6PS using a novel protocol that was conducted in a 96-well format and featured the in situ cleavage of the fusion protein to release the mutant PLC(Bc)s. Ten mutant enzymes that exhibited significant preferences toward C6PE or C6PS were selected and analyzed by steady-state kinetics to determine their specificity constants, k(cat)/K(M). The C6PS selective clones E4G, E4Q/Y56T/F66Y, and E4K/Y56V exhibited higher specificity constants toward C6PS than wt, whereas Y56T, F66Y, and Y56T/F66Y were C6PE selective and had comparable or higher specificity constants than wt for C6PE. The corresponding wt residues were singly reinserted back into the E4Q/Y56T/F66Y and E4K/Y56V mutants via site-directed mutagenesis, and the E4Q/F66Y mutant thus obtained exhibited a 10-fold higher specificity constant toward C6PS than wt, a value significantly higher than other PLC(Bc) mutants. On the basis of available data, an aromatic residue at position 66 appears important for significant catalytic activity toward all three substrates, especially C6PC and C6PE. The charge of residue 4 also appears to be a determinant of enzyme specificity as a negatively charged residue at this position endows the enzyme with C6PC and C6PE preference, whereas a polar neutral or positively charged residue results in C6PS selectivity. Replacing Tyr56 with Val, Ala, Thr, or Ser greatly reduces activity toward C6PC. Thus, the substrate specificity of PLC(Bc) can be modulated by varying three of the amino acid residues that constitute the headgroup binding pocket, and it is now apparent that this enzyme is not evolutionarily optimized to hydrolyze phospholipids with ethanolamine or serine headgroups. PMID- 12578374 TI - 3'Sulfogalactolipid binding specifically inhibits Hsp70 ATPase activity in vitro. AB - A method for the generation of soluble glycosphingolipid derivatives that retain the receptor activity of the parent (BBRC 257:391-394, Carb Res 335:91-100) was used to investigate the consequence of 3'sulfogalactolipid (SGL) specific binding within the N-terminal ATPase-containing domain of Hsc70. Sulfogalactosyl ceramide (SGC) was deacylated, and the resulting sulfogalactosylsphingosine coupled to an alpha-adamantane or a norbornane rigid hydrophobic frame. The resulting conjugate preferentially partitioned into water, as opposed to organic solvent. In the range of 100-300 microM, these conjugates inhibited the specific binding of bovine brain Hsc70 to immobilized SGLs. A similar dose-related inhibition of bovine brain Hsc70 ATPase activity was seen between 100 and 300 microM adamantylSGC (adaSGC). Adamantyl conjugates of glycolipids not bound by Hsp70s had no effect. Kinetic analysis indicated that adaSGC was a noncompetitive inhibitor of Hsc70 ATPase activity, a special case of mixed inhibition since the K(m) values were not statistically different, 0.89 +/- 0.024 microM to 0.93 +/- 0.038 microM, but the V(max) decreased from 0.20 +/- 0.012 pmol min(-1) microg( 1) to 0.15 +/- 0.016 pmol min(-1) microg(-1). A reproducible 5 min lag was observed prior to ATPase inhibition that could be eliminated by preincubation of adaSGC with Hsc70 or by adding the cochaperone Hdj-1. The dependence of ATPase inhibition on the rate of hydrolysis indicates that adaSGC binding occurs at a specific stage of the ATPase cycle. These studies identify a new mechanism for the regulation of Hsp70 ATPase activity. PMID- 12578375 TI - Regulation of phospholipase C-beta activity by phosphatidic acid: isoform dependence, role of protein kinase C, and G protein subunits. AB - Phosphatidic acid (PA) stimulates phospholipase C-beta(1) (PLC-beta(1)) activity and promotes G protein stimulation of PLC-beta(1) activity. The isoform dependence for PA regulation of PLC-beta activity as well as the role of PA in modulating regulation of PLC-beta activity by protein kinase C (PKC) and G protein subunits was determined. As compared to PLC-beta(1), the phospholipase C beta(3) (PLC-beta(3)) isoform was less sensitive to PA, requiring greater than 15 mol % PA for stimulation. PLC-beta(3) bound weakly to PA. PKC had little effect on PA stimulation of PLC-beta(3) activity. PKC, however, inhibited PA stimulation of PLC-beta(1) activity through a mechanism dependent on the mol % PA. Stimulation by 7.5 mol % PA was completely inhibited by PKC. Increasing the PA and Ca(2+) concentration attenuated PKC inhibition. The binding of PLC-beta(1) to PA containing phospholipid vesicles was also reduced by PKC, in a manner dependent on the mol % PA. PA increased the stimulation of PLC-beta(1) activity by G alpha q but had little effect on the stimulation by beta gamma subunits. These results demonstrate that PA stimulation of PLC-beta activity is tightly regulated, suggesting the existence of a distinct PA binding region in PLC beta(1). PA may be an important component of a receptor mediated signaling mechanism that determines PLC-beta(1) activation. PMID- 12578376 TI - A hydrophobicity scale for the lipid bilayer barrier domain from peptide permeabilities: nonadditivities in residue contributions. AB - Passive peptide transport across lipid membranes is governed by the energetics of partitioning into the ordered chain interior coupled with the rate of diffusion across this region. A hydrophobicity scale for peptide transfer into the barrier region of membranes derived from permeability coefficients would be useful to predict passive permeation of peptides across biomembranes and for determining the thermodynamics of peptide/protein insertion into the membrane interior. This study reports transport rates across large unilamellar vesicles (LUVs) composed of egg lecithin at 25 degrees C for a series of peptides having the general structure N-p-toluyl-(X)(n) (n =1-3), where X is glycine, alanine, or sarcosine. Apparent residue group contributions were calculated from permeability coefficients, P(RX), using the equation Delta(Delta G degrees )(X) = -RT ln(P(RX)/P(RH)). Multiple linear least-squares regression analysis performed for the set of 14 permeants yielded the best correlation (r(2) = 0.9993) when the following permeant descriptors were utilized: side-chain nonpolar surface area, number of -CONH- residues, number of toluyl-CON(Me)- residues, and number of other -CON(Me)- residues. The backbone -CONH- residue contribution in peptides, 4.6 kcal/mol, is significantly lower than that obtained for a single isolated CONH- (>6 kcal/mol), suggesting a possible influence of intramolecular hydrogen bonding. Under closer scrutiny, Delta(Delta G degrees )(X) for the Ala and Gly residues decrease with increasing peptide length. The effect of N-methylation is also highly dependent on position and number of N-methyl groups on the molecule (Delta(Delta G degrees )(X) = -0.5 to -2.2 kcal/mol). These nonadditivities may be rationalized by considering the effects of peptide length and N-methylation on membrane-induced intramolecular hydrogen bonding leading to various folded conformations. PMID- 12578377 TI - Free fatty acids modulate intermembrane trafficking of cholesterol by increasing lipid mobilities: novel 13C NMR analyses of free cholesterol partitioning. AB - Cholesterol and free fatty acids in membranes modulate major biological processes, and their cellular metabolism and actions are often coordinately regulated. However, effects of free fatty acid on cholesterol-membrane interactions have proven difficult to monitor in real time in intact systems. We developed a novel (13)C NMR method to assess effects of free fatty acids on molecular interactions of cholesterol within--and transfer between--model membranes. An important advantage of this method is the ability to acquire kinetic data without separation of donor and acceptor membranes. Large unilamellar phospholipid vesicles (LUV) with phosphatidylcholine/cholesterol ratios of 4:1 served as cholesterol donors. Small unilamellar vesicles (SUV) made with phosphatidylcholine were acceptors. The (13)C(4)-cholesterol peak is narrow in SUV, but very broad in LUV, spectra; the increase in intensity of this peak over time monitored transfer. Oleic acid and other long chain free fatty acids [saturated (C12-18) and unsaturated (C18)] dose-dependently increased mobilities of lipids in LUV (phospholipid and cholesterol) and cholesterol transfer rates, whereas short (C8-10) and very long (C24) chain free fatty acids did not. Decreasing pH from 7.4 to 6.5 (+/-oleic acid) had no effect on cholesterol transfer, and 5 mol % fatty acyl-CoAs increased transfer rates, demonstrating greater importance of the fatty-acyl tail over the headgroup. In LUV containing sphingomyelin, transfer rates decreased, but the presence of oleic acid increased transfer 1.3-fold. These results demonstrate free fatty acid-facilitated cholesterol movement within and between membranes, which may contribute to their multiple biological effects. PMID- 12578378 TI - Mitochondrial form of a tRNA synthetase can be made bifunctional by manipulating its leader peptide. AB - Previous studies showed that yeast VAS1 encodes both the cytoplasmic and mitochondrial forms of valyl-tRNA synthetase (ValRS), using alternative transcription and translation. The ValRS isoforms have identical polypeptide sequences, except for a 46-amino acid leader peptide that functions as a mitochondrial targeting signal. Although the two forms of the enzyme exhibit indistinguishable tRNA specificities in vitro, they cannot substitute for each other in vivo because of their different localizations. Here we show that the 46 residue leader sequence can be divided into two nonoverlapping peptides, each of which retains the ability to target the enzyme into mitochondria. The engineered proteins (with truncated leader sequences) are dual-targeted, rescuing both the cytoplasmic and mitochondrial defects of a vas1 knockout strain. Thus, in addition to alternative splicing and alternative translation initiation as mechanisms by which a single gene can encode cytoplasmic and mitochondrial activities, the inherent characteristics of a single polypeptide may enable it to be distributed simultaneously between two cellular compartments. This mechanism may explain how certain other single genes in Saccharomyces cerevisiae provide dual functions. PMID- 12578379 TI - Metal dependency for transcription factor rho activation. AB - The Escherichia coli rho transcription termination factor terminates select transcripts and rho activity requires Mg(2+). We investigated whether divalent metal ions other than Mg(2+) catalyze rho-dependent ATP hydrolysis to ADP and P(i) in vitro. The effects of 11 divalent metal ions (Be(2+), Ca(2+), Cd(2+), Co(2+), Cu(2+), Hg(2+), Mn(2+), Ni(2+), Sr(2+), VO(2+), Zn(2+)) on ATPase activity were determined in the absence and presence of MgCl(2). Without MgCl(2), Ca(2+), Cd(2+), Co(2+), Cu(2+), Hg(2+), Mn(2+), Ni(2+), VO(2+), and Zn(2+) activated ATP hydrolysis with either hyberbolic (Ca(2+), Co(2+), Cu(2+), Hg(2+), VO(2+)), peak velocity (Cd(2+), Mn(2+), Zn(2+)), or sigmoidal (Ni(2+)) rate acceleration curves. Sr(2+) was found to be a nonactivator and Be(2+) an inhibitor of rho-dependent ATPase activity. The metals' effects were compared with Mg(2+) and gave different rank orders when either the velocity (V(max), V(peak)) or the efficiency (V(max)/K(M), V(peak)/K(M)) of ATP hydrolysis was used as the determinant (V: Mg(2+) approximately Mn(2+) > Zn(2+) > Co(2+) > Ni(2+) approximately Cd(2+) > Ca(2+) > Cu(2+) > Hg(2+) approximately VO(2+); V/K(M): Mg(2+) > Mn(2+) > Ca(2+) > Co(2+) > Zn(2+) > Cu(2+) > Ni(2+) > Hg(2+) > Cd(2+)). Mg(2+) proved to be the most effective divalent metal. We observed that the metal dependent rates were affected by metal ion interactions with rho, RNA, and the buffer constituents. Significantly, replacement of the octahedral Mg(2+) ion by metals that typically prefer coordination spheres less than six (Cd(2+), Co(2+), Ni(2+), VO(2+), Zn(2+)) led to ATPase activity, suggesting that the putative Mg x ATP(2-) coordination sphere in rho does not need to remain fully intact for ATP hydrolysis. PMID- 12578380 TI - Correlating structure and affinity for PEX5:PTS1 complexes. AB - Many proteins that are destined to reside within the lumen of the peroxisome contain the peroxisomal targeting signal-1 (PTS1), a C-terminal tripeptide approximating the consensus sequence -Ser-Lys-Leu-COO(-). The PTS1 is recognized by the tetratricopeptide repeat (TPR) domains of PEX5, a cytosolic receptor that cycles between the cytoplasm and the peroxisome. To gain insight into the energetics of PTS1 binding specificity and to correlate these with features from the recently determined structure of a PEX5:PTS1 complex, we used a fluorescence based binding assay that enables the quantitation of the dissociation constants for PTS1-containing peptide complexes with the TPR region of human PEX5. Through application of this assay to a collection of pentapeptides containing different C terminal tripeptide sequences, including both natural and unnatural amino acids, the thermodynamic effects of sequence variation were examined. PTS1 variants that correspond to known functional targeting signals bind to the PEX5 fragment with a change in the standard binding free energy within 1.8 kcal mol(-1) of that corresponding to the peptide ending with -Ser-Lys-Leu-COO(-). The results suggest that a binding energy threshold may determine the functionality of PTS1 sequences. PMID- 12578381 TI - Lateral mobility and specific binding to GABA(A) receptors on hippocampal neurons monitored by fluorescence correlation spectroscopy. AB - The binding behavior of a fluorescently labeled muscimol derivative to the GABA(A) receptor was analyzed at rat hippocampal neurons by fluorescence correlation spectroscopy. After muscimol had been labeled with the fluorophore Alexa Fluor 532, specific binding constants for binding of the dye-labeled ligand (Mu-Alexa) to the GABA(A) receptor were determined. We found a high specific binding affinity of Mu-Alexa with a K(D) value of 3.4 +/- 0.5 nM and a rate constant of ligand-receptor dissociation (k(diss)) of (5.37 +/- 0.95) x 10(-2) s( 1). A rate constant of ligand-receptor association (k(ass)) of (1.57 +/- 0.28) x 10(7) L mol(-1) s(-1) was calculated. The following diffusion coefficients were observed: D(free) = 233 +/- 20 microm(2)/s (n = 66) for free diffusing Mu-Alexa, D(bound1) = 2.8 +/- 0.9 microm(2)/s (n = 64) for the lateral mobility, and D(bound2) = 0.14 +/- 0.05 microm(2)/s (n = 56) for the hindered mobility of the GABA(A) receptor-ligand complex in the cell membrane. Saturation of Mu-Alexa binding was observed at a concentration of 50 nM. A maximum number of binding sites [B(max) = 18.4 +/- -0.4 nM (n = 5)] was found. Similar K(i) values of 4.5 +/- 1.0 nM for nonlabeled muscimol and 8.8 +/- 1.8 nM for Mu-Alexa were found by RRAs using [(3)H]muscimol as a radioligand. A concentration-dependent increase in the level of specific Mu-Alexa binding was demonstrated by the positive cooperative activity of co-incubated midazolam, which was selectively found in GABA(A) receptor-ligand complexes with hindered mobility. PMID- 12578382 TI - Thiols as classical and slow-binding inhibitors of IMP-1 and other binuclear metallo-beta-lactamases. AB - The inhibitory effect of a variety of thiol compounds on the function of binuclear metallo-beta-lactamases, with a particular focus on IMP-1 from Pseudomonas aeruginosa, has been investigated. Thiol inhibitors, depending on their structural features, fall into two categories, one in which inhibition at neutral pH was instantaneous and the other in which inhibition was time dependent. While mercaptans with anionic substituents in the vicinity of their SH groups exhibited the former type of inhibition, neutral thiols appear to induce a slow, time-dependent isomerization of the initially formed EI complex to a tighter EI complex. Kinetic parameters describing the latter process were obtained by fitting progress curves of substrate hydrolysis using standard and numerical procedures. The failure of charged thiols to exhibit slow binding is suggested to be due to a rapid isomerization of the initial EI complex. Slow binding in the case of neutral thiols was observed only below pH 8. Studies on the pH dependence of catalysis by IMP-1 revealed that (i) enzyme inactivation at low pH is a slow process with presumably two groups with a pK(a) of approximately 5.2 in the protein being responsible for the loss of activity, (ii) inhibition by thiols is independent of pH between pH 5 and 9, and (iii) an apparent enhancement of the catalytic activity of IMP-1 by thiols occurs at pH <5. The last mentioned phenomenon is explained by a model in which mercaptans retard the proton dependent isomerization of the enzyme. Studies on the thiol-mediated inhibition of the binuclear forms of Bacteroides fragilis (CcrA) and Bacillus cereus (BcII strain 5/B/6) metallo-beta-lactamase have revealed that while CcrA was instantaneously albeit moderately inhibited by mercaptans, BcII mimicked IMP-1 in its interaction with thiols. These differences are proposed to be due partly to the structural divergence of these proteins in the vicinity of Zn2. PMID- 12578383 TI - Conformational and thermodynamic characterization of the molten globule state occurring during unfolding of cytochromes-c by weak salt denaturants. AB - The denaturation of bovine and horse cytochromes-c by weak salt denaturants (LiCl and CaCl(2)) was measured at 25 degrees C by observing changes in molar absorbance at 400 nm (Delta epsilon(400)) and circular dichroism (CD) at 222 and 409 nm. Measurements of Delta epsilon(400) and mean residue ellipticity at 409 nm ([theta](409)) gave a biphasic transition for both modes of denaturation of cytochromes-c. It has been observed that the first denaturation phase, N (native) conformation <--> X (intermediate) conformation and the second denaturation phase, X conformation <--> D (denatured) conformation are reversible. Conformational characterization of the X state by the far-UV CD, 8-anilino-1 naphthalene sulfonic acid (ANS) binding, and intrinsic viscosity measurements led us to conclude that the X state is a molten globule state. Analysis of denaturation transition curves for the stability of different states in terms of Gibbs energy change at pH 6.0 and 25 degrees C led us to conclude that the N state is more stable than the X state by 9.55 +/- 0.32 kcal mol(-1), whereas the X state is more stable than the D state by only 1.40 +/- 0.25 kcal mol(-1). We have also studied the effect of temperature on the equilibria, N conformation <- > X conformation and X conformation <--> D conformation in the presence of different denaturant concentrations using two different optical probes, namely, [theta](222) and Delta epsilon(400). These measurements yielded T(m), (midpoint of denaturation) and Delta H(m) (enthalpy change) at T(m) as a function of denaturant concentration. A plot of Delta H(m) versus corresponding T(m) was used to determine the constant-pressure heat capacity change, Delta C(p) (= ( partial differential Delta H(m)/ partial differential T(m))(p)). Values of Delta C(p) for N conformation <--> X conformation and X conformation <--> D conformation is 0.92 +/- 0.02 kcal mol(-1) K(-1) and 0.41 +/- 0.01 kcal mol(-1) K(-1), respectively. These measurements suggested that about 30% of the hydrophobic groups in the molten globule state are not accessible to the water. PMID- 12578384 TI - Comprehensive model for allosteric regulation of mammalian ribonucleotide reductase: refinements and consequences. AB - Reduction of NDPs by murine ribonucleotide reductase (mRR) requires catalytic (mR1) and free radical-containing (mR2) subunits and is regulated by nucleoside triphosphate allosteric effectors. Here we present the results of several studies that refine the recently presented comprehensive model for the allosteric control of mRR enzymatic activity [Kashlan, O. B., et al. (2002) Biochemistry 41, 462 474], in which nucleotide binding to the specificity site (s-site) drives formation of an active R1(2)R2(2) dimer, ATP or dATP binding to the adenine site (a-site) drives formation of a tetramer, mR1(4a), which isomerizes to an inactive form, mR1(4b), and ATP binding to the hexamerization site (h-site) drives formation of an active R1(6)R2(6) hexamer. Analysis of the a-site D57N variant of mR1, which differs from wild-type mR1 (wt-mR1) in that its RR activity is activated by both ATP and dATP, demonstrates that dATP activation of the D57N variant RR arises from a blockage in the formation of mR1(4b) from mR1(4a), and provides strong evidence that mR1(4a) forms active complexes with mR2(2). We further demonstrate that (a) differences in the effects of ATP versus dATP binding to the a-site of wt-mR1 provide specific mechanisms by which the dATP/ATP ratio in mammalian cells could modulate in vivo RR enzymatic activity, (b) the comprehensive model is valid over a range of Mg(2+) concentrations that include in vivo concentrations, and (c) equilibrium constants derived for the comprehensive model can be used to simulate the distribution of R1 among dimer, tetramer, and hexamer forms in vivo. Such simulations indicate that mR1(6) predominates over mR1(2) in the cytoplasm of normal mammalian cells, where the great majority of RR activity is located, but that mR1(2) may be important for nuclear RR activity and for RR activity in cells in which the level of ATP is depleted. PMID- 12578385 TI - Crystallization of beta-galactosidase does not reduce the range of activity of individual molecules. AB - By use of a capillary electrophoresis-based procedure, it is possible to measure the activity of individual molecules of beta-galactosidase. Molecules from the crystallized enzyme as well as the original enzyme preparation used to grow the crystals both displayed a range of activity of 20-fold or greater. beta Galactosidase molecules obtained from two different crystals had indistinguishable activity distributions of 31,600 +/- 1100 and 31,800 +/- 1100 reactions min(-1) (enzyme molecule)(-1). This activity was found to be significantly different from that of the enzyme used to grow the crystals, which showed an activity distribution of 38,500 +/- 900 reactions min(-1) (enzyme molecule)(-1). PMID- 12578386 TI - Substitutions for glutamate 101 in subunit II of cytochrome c oxidase from Rhodobacter sphaeroides result in blocking the proton-conducting K-channel. AB - Two functional input pathways for protons have been characterized in the heme copper oxidases: the D-channel and the K-channel. These two proton-conducting channels have different functional roles and have been defined both by X-ray crystallography and by the characterization of site-directed mutants. Whereas the entrance of the D-channel is well-defined as D132(I) (subunit I; Rhodobacter sphaeroides numbering), the entrance of the K-channel has not been clearly defined. Previous mutagenesis studies of the cytochrome bo(3) quinol oxidase from Escherichia coli implicated an almost fully conserved glutamic acid residue within subunit II as a likely candidate for the entrance of the K-channel. The current work examines the properties of mutants of this conserved glutamate in the oxidase from R. sphaeroides (E101(II)I,A,C,Q,D,N,H) and residues in the immediate vicinity of E101(II). It is shown that virtually any substitution for E101(II), including E101(II)D, strongly reduces oxidase turnover (to 8-29%). Furthermore, the low steady-state activity correlates with an inhibition of the rate of reduction of heme a(3) prior to the reaction with O(2). These are phenotypes expected of K-channel mutants. It is concluded that the predominant entry point for protons going into the K-channel of cytochrome oxidase is the surface-exposed glutamic acid E101(II) in subunit II. PMID- 12578387 TI - Quinone reduction via secondary B-branch electron transfer in mutant bacterial reaction centers. AB - Symmetry-related branches of electron-transfer cofactors-initiating with a primary electron donor (P) and terminating in quinone acceptors (Q)-are common features of photosynthetic reaction centers (RC). Experimental observations show activity of only one of them-the A branch-in wild-type bacterial RCs. In a mutant RC, we now demonstrate that electron transfer can occur along the entire, normally inactive B-branch pathway to reduce the terminal acceptor Q(B) on the time scale of nanoseconds. The transmembrane charge-separated state P(+)Q(B)(-) is created in this manner in a Rhodobacter capsulatus RC containing the F(L181)Y Y(M208)F-L(M212)H-W(M250)V mutations (YFHV). The W(M250)V mutation quantitatively blocks binding of Q(A), thereby eliminating Q(B) reduction via the normal A branch pathway. Full occupancy of the Q(B) site by the native UQ(10) is ensured (without the necessity of reconstitution by exogenous quinone) by purification of RCs with the mild detergent, Deriphat 160-C. The lifetime of P(+)Q(B)(-) in the YFHV mutant RC is >6 s (at pH 8.0, 298 K). This charge-separated state is not formed upon addition of competitive inhibitors of Q(B) binding (terbutryn or stigmatellin). Furthermore, this lifetime is much longer than the value of approximately 1-1.5 s found when P(+)Q(B)(-) is produced in the wild-type RC by A side activity alone. Collectively, these results demonstrate that P(+)Q(B)(-) is formed solely by activity of the B-branch carriers in the YFHV RC. In comparison, P(+)Q(B)(-) can form by either the A or B branches in the YFH RC, as indicated by the biexponential lifetimes of approximately 1 and approximately 6-10 s. These findings suggest that P(+)Q(B)(-) states formed via the two branches are distinct and that P(+)Q(B)(-) formed by the B side does not decay via the normal (indirect) pathway that utilizes the A-side cofactors when present. These differences may report on structural and energetic factors that further distinguish the functional asymmetry of the two cofactor branches. PMID- 12578388 TI - Energy and electron transfer in photosystem II of a chlorophyll b-containing Synechocystis sp. PCC 6803 mutant. AB - Using a Synechocystis sp. PCC 6803 mutant strain that lacks photosystem (PS) I and that synthesizes chlorophyll (Chl) b, a pigment that is not naturally present in the wild-type cyanobacterium, the functional consequences of incorporation of this pigment into the PS II core complex were investigated. Despite substitution of up to 75% of the Chl a in the PS II core complex by Chl b, the modified PS II centers remained essentially functional and were able to oxidize water and reduce Q(A), even upon selective excitation of Chl b at 460 nm. Time-resolved fluorescence decay measurements upon Chl excitation showed a significant reduction in the amplitude of the 60-70 ps component of fluorescence decay in open Chl b-containing PS II centers. This may indicate slower energy transfer from the PS II core antenna to the reaction center pigments or slower energy trapping. Chl b and pheophytin b were present in isolated PS II reaction centers. Pheophytin b can be reversibly photoreduced, as evidenced from the absorption bleaching at approximately 440 and 650 nm upon illumination in the presence of dithionite. Upon excitation at 685 nm, transient absorption measurements using PS II particles showed some bleaching at 650 nm together with a major decrease in absorption around 678 nm. The 650 nm bleaching that developed within approximately 10 ps after the flash and then remained virtually unchanged for up to 1 ns was attributed to formation of reduced pheophytin b and oxidized Chl b in some PS II reaction centers. Chl b-containing PS II had a lower rate of charge recombination of Q(A)(-) with the donor side and a significantly decreased yield of delayed luminescence in the presence of DCMU. Taken together, the data suggest that Chl b and pheophytin b participate in electron-transfer reactions in PS II reaction centers of Chl b-containing mutant of Synechocystis without significant impairment of PS II function. PMID- 12578389 TI - Unfolding of class A amphipathic peptides on a lipid surface. AB - The folding of polypeptides associated with biomembranes is a ubiquitous phenomenon, yet the thermodynamics underlying the process are poorly understood. In the present work we examine the unfolding of a series of alpha-helical amphipathic membrane-associated peptides using guanidine hydrochloride as a denaturant. The peptides are based on the class A amphipathic helix motif, and each contains a single tryptophan at sequence position 2, 3, 7, 12, or 14. The isothermal unfolding process was monitored by circular dichroism ellipticity at 222 nm to monitor changes in the helical structure of the peptide. Tryptophan fluorescence was used to probe the local changes in the environment about the indole fluorophore. The unfolding curves generated from the two experimental techniques for each peptide-lipid complex were non-coincidental, suggesting the presence of stable intermediate(s) in the unfolding. A three-state model could adequately account for the data and yielded parameters which were consistent with the presence of a partially folded intermediate structure which (i) is closer in Gibb's free energy to the folded state than the unfolded state and (ii) retains much of the interfacial and amphipathic character of the folded state. Denaturant induced peptide dissociation from the peptide-lipid complexes was found to be negligible as confirmed by size exclusion chromatography. The results are compared with related thermodynamic data and discussed in terms of current models of peptide folding at membrane interfaces. PMID- 12578390 TI - Real-time monitoring of the interactions of two-stranded de novo designed coiled coils: effect of chain length on the kinetic and thermodynamic constants of binding. AB - We have de novo designed a heterodimeric coiled-coil formed by two peptides as a capture/delivery system that can be used in applications such as affinity tag purification, immobilization in biosensors, etc. The two strands are designated as K coil (KVSALKE heptad sequence) and E coil (EVSALEK heptad sequence), where positively charged or negatively charged residues occupy positions e and g of the heptad repeat. In this study, for each E coil or K coil, three peptides were synthesized with lengths varying from three to five heptads. The effect of the chain length of each partner upon the kinetic and thermodynamic constants of interaction were determined using a surface plasmon resonance-based biosensor. Global fitting of the interactions revealed that the E5 coil interacted with the K5 coil according to a simple binding model. All the other interactions involving shorter coils were better described by a more complex kinetic model involving a rate-limiting reorganization of the coiled-coil structure. The affinities of these de novo designed coiled-coil interactions were found to range from 60 pM (E5/K5) to 30 microM (E3/K3). From these K(d) values, we were able to determine the free energy contribution of each heptad, depending on its relative position within the coiled-coils. We found that the free energy contribution of a heptad occupying a central position was 3-fold higher than that of a heptad at either end of the coiled-coil. The wide range of stabilities and affinities for the E/K coil system provides considerable flexibility for protein engineering and biotechnological applications. PMID- 12578391 TI - Proposed mechanism and functional amino acid residues of malonyl-CoA:anthocyanin 5-O-glucoside-6'''-O-malonyltransferase from flowers of Salvia splendens, a member of the versatile plant acyltransferase family. AB - The versatile plant acyltransferase (VPAT) family is a recently identified protein family consisting of acyltransferases involved in secondary metabolism in plants along with numerous homologues with as yet unidentified biochemical functions. Malonyl-CoA:anthocyanin 5-O-glucoside-6' "-O-malonyltransferase of Salvia splendens flowers (Ss5MaT1) is a member of this family that catalyzes the regiospecific transfer of the malonyl group from malonyl-CoA to the 6' "-hydroxyl group of the 5-glycosyl moiety of anthocyanins. To elucidate the mechanism and functional amino acid residues of VPAT family enzymes, steady-state kinetic analyses and site-directed mutagenesis of Ss5MaT1 guided by sequence comparison studies were carried out. On the basis of the results of product and dead-end inhibition studies as well as sequence comparison studies, the kinetic mechanism of Ss5MaT1 could be most consistently described in terms of a ternary complex mechanism in which both substrates and the enzyme form a complex before catalysis can occur, as in the case of chloramphenicol O-acetyltransferase (CAT) and histone acetyltransferase (HAT). Eight polar or ionizable amino acid residues that are invariant among 12 VPAT family enzymes were replaced by alanine, and the mutant enzymes were kinetically characterized. A significant diminution of the k(cat) value was observed with the substitution of His167 (relative k(cat), 0.02%) and Asp390 (<0.01%), strongly suggesting that His167 and Asp390 are very important for catalytic activity. The log k(cat) versus pH plots of the Ss5MaT1 catalyzed malonyl transfer suggested that a deprotonated active site group of pK(a) = 7.0 +/- 0.1 may be involved in the catalytic steps of the "substrate to product" conversion in the ternary enzyme-substrate complex. Taking these lines of evidence together with the suggested similarity of the kinetic and catalytic mechanisms of Ss5MaT1 to those of CAT and HAT, the following Ss5MaT1 mechanism based on general acid/base catalysis was proposed: in the ternary complex, a general base deprotonates the 6' "-hydroxyl group of the anthocyanin substrate, thereby promoting a nucleophilic attack on the carbonyl of the thioester of malonyl-CoA; His167 and Asp390 appear to be involved in the general acid/base mechanism of Ss5MaT1. PMID- 12578392 TI - Effects of engineering uphill electron transfer into the methylamine dehydrogenase-amicyanin-cytochrome c-551i complex. AB - Within the methylamine dehydrogenase-amicyanin-cytochrome c-551i complex, electrons are transferred from tryptophan tryptophylquinone (TTQ) to heme via the type I copper center of amicyanin. Mutation of Pro94 of amicyanin to Phe increases the redox potential of the copper center within the protein complex by approximately 195 mV. This introduces a large energy barrier for the second electron transfer (ET) step in this three-protein ET chain. As a consequence of this mutation, the ET rate from TTQ to copper exhibits about a 6-fold increase and the ET rate from copper to heme exhibits about a 100-fold decrease. These changes in ET rate are consistent with the predictions of Marcus theory. Temperature dependence studies of these reactions indicate that the reorganization energies for the ET to and from the copper center are unchanged by the P94F mutation, despite the large change in redox potential that it causes. Steady-state kinetic studies indicate that despite the large energy barrier for the ET from copper to heme, methylamine-dependent reduction of heme by the three protein complex with P94F amicyanin goes to completion. The turnover number for this steady-state reaction, however, is decreased 50-fold relative to that of the native complex. As a consequence of the P94F mutation, the rate constant for the unfavorable uphill ET reaction from copper to heme has become the rate-limiting step in the overall reaction. The evolutionary implications of the effects of this mutation on the function of this naturally occurring simple ET chain are discussed. PMID- 12578393 TI - Translesion synthesis past platinum DNA adducts by human DNA polymerase mu. AB - DNA polymerase mu (pol mu) is a member of the pol X family of DNA polymerases, and it shares a number of characteristics of both DNA polymerase beta (pol beta) and terminal deoxynucleotidyl transferase (TdT). Because pol beta has been shown to perform translesion DNA synthesis past cisplatin (CP)- and oxaliplatin (OX)-GG adducts, we determined the ability of pol mu to bypass these lesions. Pol mu bypassed CP and OX adducts with an efficiency of 14-35% compared to chain elongation on undamaged DNA, which is second only to pol eta in terms of bypass efficiency. The relative ability of pol mu to bypass CP and OX adducts was dependent on both template structure and sequence context. Since pol mu has been shown to be more efficient on gapped DNA templates than on primed single-stranded DNA templates, we determined the ability of pol mu to bypass Pt-DNA adducts on both primed single-stranded and gapped templates. The bypass of Pt-DNA adducts by pol mu was highly error-prone on all templates, resulting in 2, 3, and 4 nt deletions. We postulate that bypass of Pt-DNA adducts by pol mu may involve looping out the Pt-GG adduct to allow chain elongation downstream of the adduct. This reaction appears to be facilitated by the presence of a downstream "acceptor" and a gap large enough to provide undamaged template DNA for elongation past the adduct, although gapped DNA is clearly not required for bypass. PMID- 12578394 TI - Purification and properties of the Dictyostelium calpain-like protein, Cpl. AB - Calpains are intracellular, cysteine proteases found in plants, animals, and fungi. There is emerging evidence that they are important mediators of cell adhesion and motility in animal cells. Because the cellular slime mold, Dictyostelium discoideum, is a genetically tractable model for cell adhesion and motility, we have investigated whether a calpain-like protein is expressed in this organism. Contig 13130 (Sanger Institute Dictyostelium sequencing project) was identified as a three-exon gene that encodes a calpain-like protein. Using a custom peptide antibody to assay for the presence of this putative protein, we identified Dictyostelium calpain-like protein (Cpl) and purified it to near homogeneity. Cpl is a 72278 Da cytosolic protein. Weak caseinolytic activity inhibitable by cysteine protease inhibitors was copurified with Cpl immunoreactivity, and purified Cpl appeared to undergo autoproteolysis upon transfer to inhibitor-free buffer. The major cleavage, generating a 51291 Da form, occurred after Pro 189. The Cpl domain structure resembles mammalian calpain 10, comprising an N-terminal catalytic domain followed by tandem calpain D-III domains. The putative catalytic domain appears to possess His and Gln active site residues, instead of the canonical His and Asn residues in calpains. The active site Cys has not yet been identified, and definitive proof of a proteolytic function awaits further study. Its phylogenetic distribution in D. discoideum and several protists suggests that the calpain D-III domain evolved early in eukaryotic cells. PMID- 12578395 TI - Trp-999 of beta-galactosidase (Escherichia coli) is a key residue for binding, catalysis, and synthesis of allolactose, the natural lac operon inducer. AB - Trp-999 is a key residue for the action of beta-galactosidases (Escherichia coli). Several site specific substitutions (Phe, Gly, Tyr, Leu) for Trp-999 were made. Each substitution caused greatly decreased affinities for substrates and inhibitors that bind in the "shallow" mode, while the affinities of inhibitors that bind in the "deep" mode were not decreased nearly as much. This shows that Trp-999 is important for binding in the shallow mode. The residue is also very important for binding glucose to galactosyl-beta-galactosidase (as a transgalactosidic acceptor). Substitution greatly diminished the affinity for glucose. Substitutions also changed the activation thermodynamics and, subsequently, the rates of the catalytic reactions. The enthalpies of activation of the glycolytic bond cleavage step (galactosylation, k(2)) became less favorable while the entropies of activation of that step became more favorable as a result of the substitutions. Differing magnitudes of these enthalpic and entropic effects with ONPG as compared to PNPG caused the k(2) values for ONPG to decrease but to increase for PNPG. The enthalpies of activation for the common hydrolytic step (degalactosylation, k(3)) increased while the entropies of activation for this step did not change much. As a result, k(3) became small and rate determining for each substituted enzyme. The substitutions caused the rate constant (k(4)) of the transgalactosidic acceptor reactions with glucose (for the formation of allolactose) to become much larger and of the same order of magnitude as the normally large rate constants for transgalactosidic acceptor reactions with small alcohols. This is probably because glucose can approach with less restriction in the absence of Trp-999. However, since glucose binds very poorly to the galactosyl-beta-galactosidases with substitutions for Trp-999, the proportion of lactose molecules converted to allolactose is small. Thus, Trp-999 is also important for ensuring that an appropriate proportion of lactose is converted to allolactose. PMID- 12578397 TI - Influenza vaccination and antiviral therapy: is there a role for concurrent administration in the institutionalised elderly? AB - Influenza vaccination is estimated to be 50-68% efficacious in preventing pneumonia, hospitalisation or death in nursing home residents. Large culture proven outbreaks may occur despite high resident vaccination rates. There is, therefore, a significant role for concurrent administration of influenza vaccination and antiviral therapy. The use of antiviral treatment and chemoprophylaxis requires community reporting of viral isolates, and contingency plans for rapid case identification and application of antiviral therapy. Clinicians must react quickly to control a highly infectious seasonal pathogen that may strike as an explosive outbreak. This situation is unique in geriatric practice. Current antiviral treatment should be administered within 48 hours of symptom onset, and is more efficacious if administered within 12 hours. In the case of an explosive institutional outbreak, a 1-day delay in prophylaxis may allow infection of many residents with a potentially fatal illness. Influenza must be differentiated from other respiratory viruses or syndromes. Grouped rapid diagnostic tests can aid laboratory confirmation. Antiviral agents include the M(2) inhibitors, amantadine and rimantadine, active against influenza A, and the neuraminidase inhibitors, zanamivir and oseltamivir, active against influenza A and B. In our experience, influenza B illness is as severe as influenza A. All agents have similar efficacy as treatment and prophylaxis against sensitive strains. When M(2) inhibitors are used simultaneously within an enclosed space (i.e. household or nursing home) as both treatment and prophylaxis, resistant strains may emerge that limit prophylactic efficacy. When M(2) inhibitors are administered to suspected cases (residents or staff) in institutions, precautions against secretion are especially important to diminish the risk of transmission of resistant virus. Rimantadine has been shown to have significantly fewer CNS adverse events compared with amantadine. Amantadine and oseltamivir require dosage adjustment in those with renal impairment. Oseltamivir, rimantadine and amantadine are administered by mouth, while zanamivir is administered by oral inhalation in a lactose powder. The labelling advises caution in the use of zanamivir in those with underlying airway disease. Pooled analysis of studies in patients given zanamivir indicate that individuals over the age of 50 years (at high risk for complications) and those severely symptomatic at presentation, tend to benefit most from early treatment. Neuraminidase inhibitors also diminish the need for antibacterials to treat secondary complications. An institutional programme to control influenza should include vaccination, and contingency plans for clinical surveillance, specimen processing and the rapid application of antiviral treatment and prophylaxis. PMID- 12578398 TI - Bisphosphonates to prevent osteoporosis in men receiving androgen deprivation therapy for prostate cancer. AB - Osteoporosis is an important complication of androgen deprivation therapy for prostate cancer. Androgen deprivation therapy either by bilateral orchiectomies or treatment with a gonadotropin-releasing hormone agonist decreases bone mineral density (BMD) and increases the risk of fracture. Dietary factors and lifestyle may contribute to bone loss. There are limited prospective data about treatment or prevention of osteoporosis in men with prostate cancer and many recommendations are based on studies of postmenopausal osteoporosis. Lifestyle modification including smoking cessation, moderation of alcohol consumption, and regular weight bearing exercise should be encouraged. Supplemental calcium and vitamin D are recommended. Additional treatment may be warranted for men with osteoporosis, fractures, or high rates of bone loss during androgen deprivation therapy. Recent studies have evaluated the efficacy of bisphosphonates to prevent bone loss during androgen deprivation therapy. Pamidronate (pamidronic acid), a second-generation bisphosphonate, prevents bone loss in the hip and spine during androgen deprivation therapy. Zoledronic acid, a more potent third-generation bisphosphonate, not only prevents bone loss but also increases BMD in the hip and spine. Alendronate (alendronic acid) is approved for the treatment of osteoporosis in men although its efficacy and that of other oral bisphosphonates has not been evaluated in men receiving androgen deprivation therapy. Additional prospective studies are needed to evaluate the long-term effects of bisphosphonates on fracture risk and disease-related outcomes. PMID- 12578399 TI - Early treatment of benign prostatic hyperplasia: implications for reducing the risk of permanent bladder damage. AB - A significant change has occurred in the management of symptomatic benign prostatic hyperplasia (BPH) since effective pharmacological treatment became available and led to a significant decrease in the number of surgical procedures in many Western countries. The hypothesis of a causative role of benign prostatic enlargement and bladder outflow obstruction (BOO) in lower urinary tract symptoms (LUTS) was based on the association between prostate growth and symptoms of prostatism in elderly men and on the dramatic reduction of LUTS upon relief of obstruction. Careful investigation into the epidemiology of LUTS and BPH failed to confirm such an association and opened new perspectives in the pathophysiology of lower urinary tract dysfunction and symptoms. The observation that LUTS were equally distributed in male and female cohorts, when matched for age, moved attention away from the prostate and towards the urinary bladder and its aging related disorders. When BPH surgery was developed, the management of the disease was aimed at preventing death from chronic renal failure, but the picture has changed and modern medical treatment is now aimed at improving the patient's quality of life. The increasing size of elderly populations in the Western world and the consequent financial constraints of national healthcare systems have raised the question of when pharmacological treatment of symptomatic BPH should be initiated. Retrospective and prospective analysis of various BPH populations and clinical studies has clearly defined the capacity of pharmacological treatment to reduce the incidence of complications of BPH, such as acute urinary retention and the need for surgery, but the cost/benefit ratio is unclear. Notwithstanding the limitations inherent in the experimental models, there is evidence from various animal models, investigating the pathophysiology of the urinary bladder in the presence of outflow obstruction, to indicate that a cause and effect relationship between BOO and bladder decompensation has been established and to support the hypothesis that permanent bladder damage may occur when the obstruction is not relieved early enough. Preliminary experimental evidence also suggests that alpha(1)-adrenoceptor antagonists may have a role in reducing the damaging effects of BOO on the urinary bladder. At present, there is no evidence to support the need for early pharmacological treatment of symptomatic BPH with no BOO beyond the obvious target of improving the patient's quality of life. The evidence for early treatment of BOO and the need to preserve bladder function is clear. Further experimental and clinical research is required to identify markers of early bladder damage and decompensation which can be used to select patients for early pharmacological treatment of BPH. PMID- 12578401 TI - Treatment of chronic obstructive pulmonary disease in older patients: a practical guide. AB - Chronic obstructive pulmonary disease (COPD) is a common disability, largely encountered in the elderly population, in whom it causes significant morbidity and mortality. The general perception of health professionals is that COPD is often a self-inflicted disorder affecting the more socio-economically disadvantaged segment of the population with significant comorbidity. COPD is the least funded in terms of research in relation to illness burden compared with other chronic diseases. However, recently published guidelines of both the British Thoracic Society and the Global Initiative for Chronic Obstructive Lung Disease have highlighted best management strategies both of chronic symptoms and acute exacerbations in this patient group. The chronic management of COPD should, like asthma, involve a stepwise approach with smoking cessation being pivotal for all severities of COPD, regardless of patient age. The mainstay of therapeutic treatment remains regular bronchodilators, both beta(2)-adrenoreceptor agonists and anticholinergic agents. Current evidence suggests that long-acting beta(2) adrenoreceptor agonists such as salmeterol and the new long-acting anticholinergic agent tiotropium bromide are more efficacious than their shorter acting equivalents such as salbutamol and ipratropium bromide in terms of bronchodilation, improved well-being and a reduction in acute exacerbation rates. Additionally since they are taken once or twice daily compliance should be improved. The role of long-term inhaled corticosteroids in the chronic management of COPD is contentious. Only those patients with COPD who have been shown to respond to a formal corticosteroid trial, preferably with a 2-week course of oral corticosteroid, should receive long-term inhaled corticosteroids. In the management of acute exacerbations in acidotic patients nasal ventilation is the treatment of choice in addition to conventional treatment with bronchodilators and oral corticosteroids. Antibacterials need not be prescribed universally in all exacerbations of COPD. Pulmonary rehabilitation classes either individually or in groups have been shown to be beneficial in the management of patients with COPD and their use in secondary care is to be encouraged. Most treatment modalities do not improve pulmonary function in patients with severe COPD. Therefore, pulmonary function including spirometry should be used to make the diagnosis of COPD but not as a monitor of efficacy of treatment. Assessment of severity of COPD and improvement with treatment modalities is best done with dynamic exercise testing such as 6-minute walk tests and incremental shuttle walk tests or with the administration of disease-specific physical disability and quality-of-life questionnaires. Most COPD research does not specifically target the older COPD patients and these patients may merit special consideration for their optimum assessment and management. PMID- 12578403 TI - Division of Surgery, Royal North Shore Hospital, Sydney, Australia. PMID- 12578400 TI - Treatment of pruritus associated with systemic disorders in the elderly: a review of the role of new therapies. AB - Generalised pruritus is common in the elderly. Idiopathic 'senile pruritus' is a diagnosis of exclusion, and an underlying systemic disorder should be sought. Thyroid disease, haematological malignancy, iron deficiency, cholestasis or renal impairment may be responsible for pruritus. Rarely pruritus may occur after cerebral infarction or as a paraneoplastic phenomenon. The mechanisms of pruritus are poorly understood. In systemic disorders, correction of the underlying disorder alleviates itch. However, when this cannot be achieved, a symptomatic approach is required. Response to treatment varies enormously and an empirical approach is often required. Topical applications are available to soothe the skin and bandaging techniques may improve their efficacy. A number of more targeted treatments are available for renal and cholestatic pruritus. Novel therapies such as thalidomide, opioid antagonists, ondansetron and phototherapy with ultraviolet (UV)-B radiation are now being used. Treatment of pruritus needs to be individualised, and the elderly present a particular challenge. Adequate delivery of simple emollients may be impossible because of physical impairment The elderly are more vulnerable to the adverse effects of treatments, comorbidities may alter the pharmacokinetics of drug metabolism and polypharmacy increases the likelihood of adverse drug interactions. Cognitive impairment can lead to poor compliance with treatment. The patient's general health, the severity of symptoms and the adverse effects of treatment all need to be considered. Most treatments are of benefit only to some patients; others derive only marginal improvement. Many of the newer treatments are unlicensed for pruritus and should preferably be administered under specialist supervision. We review the literature concerning the treatment of itch associated with systemic diseases, with particular emphasis on issues relevant to the elderly. Pruritus is a difficult symptom to treat. However, it is hoped that research into the mechanisms underlying the pruritus of systemic disease will allow a better understanding so that we should be able to look forward to more specific and effective therapies in the future. PMID- 12578402 TI - Interventions to improve medication compliance in older patients living in the community: a systematic review of the literature. AB - Noncompliance affects all age groups, but older patients have specific barriers against effective medication use and can be more vulnerable to the incorrect use of medication. Some age-related barriers are vision loss and cognitive impairment. In people aged 60 years or older, noncompliance with medication regimens varies from 26-59%. In this article we review randomised controlled trials (RCTs) on interventions aimed to improve compliance with medication regimens in older patients living in the community. It is known from other reviews that multifaceted interventions, a combination of interventions, are more often effective than single-focus interventions, probably because these address a wider range of barriers. Also, an individual approach with specifically tailored interventions is effective at improving compliance. In this review the following two hypotheses are evaluated: i) multifaceted interventions improve compliance more successfully than single interventions; and ii) interventions that are tailored to patients improve compliance more successfully than generalised interventions. A systematic literature search in four databases produced 14 suitable RCTs with 23 interventions that we categorised as single or multifaceted, and generalised or tailored. Differences in medication compliance between the intervention group and the control group were in less than half of the comparisons in favour of the intervention group. Telephone-linked reminder systems achieved the most striking effect, looking at differences between the intervention and control group, and deserve further attention. Overall, multifaceted interventions and tailored interventions seemed to result more often in differences in compliance rates in older adults in favour of the intervention group compared with a control group than single and generalised interventions. PMID- 12578404 TI - Modulation of inflammatory and catabolic responses in severely burned children by early burn wound excision in the first 24 hours. AB - HYPOTHESIS: Early burn wound excision modulates the hypermetabolic response in severe pediatric burn injuries. DESIGN: Before-after trial. SETTING: A 30-bed burn referral center in a private, university-affiliated hospital. METHODS: We studied 35 severely burned children who were divided into 2 groups. One group (n = 20) was treated with early burn wound excision within 24 hours after the injury. The second group (n = 15) was treated conservatively with silver sulfadiazine in other burn facilities for 5 days, and burn wounds were surgically excised when patients were admitted to our burn center on day 6 after the injury. Data compiled included oxygen consumption and acute-phase protein, interleukin 1beta; interleukin 6, interleukin 10, tumor necrosis factor alpha, and anabolic hormone (growth hormone, insulinlike growth factor type 1) levels preoperatively and 24 hours and 5 days postoperatively. MAIN OUTCOME MEASURES: Acute-phase and hypermetabolic responses. RESULTS: Early burn wound excision abrogated the hypermetabolic response in pediatric burn patients. Patients who underwent conservative treatment had a significantly more severe inflammatory and hypermetabolic response at the same time interval and significantly lower levels of anabolic hormones. CONCLUSIONS: Early burn wound excision is a safe therapeutic approach that modulates the hypermetabolic response after burn injury. It was superior to the conservative treatment of silver sulfadiazine and delayed excision, and it should be considered when treating all severe full thickness burns. PMID- 12578405 TI - Hand-assisted laparoscopic surgery. AB - HYPOTHESIS: Hand-assisted laparoscopic surgery (HALS) has been proposed as a useful alternative to conventional open or laparoscopic surgery. However, most information is fragmented and comes from specific or selective indications. To assess the current situation of HALS, a general overview of the fields of application, results, and quality of the evidence of these results is necessary. DATA SOURCES: Current English-language literature review. STUDY SELECTION: Case reports, series, and opinion articles on HALS. DATA EXTRACTION AND SYNTHESIS: Evaluation of the type of study and results. Most of the articles are short case series. Only a few comparative or randomized comparative trials on HALS for splenectomy and colectomy have been published. CONCLUSIONS: Hand-assisted laparoscopic surgery seems to be a promising technique that has been applied with success in a wide range of digestive tract-related surgical procedures. The main role is to help in difficult cases before conversion is necessary or for training unskilled surgeons, and not as an alternative to pure laparoscopic surgery. However, not enough evidence-based data are available to know exactly the final outcome of this technique in general surgery. Prospective randomized trials with established open or laparoscopic procedures are lacking, and these trials are needed to support the final role of HALS. PMID- 12578407 TI - Morbidity and mortality of children following motor vehicle crashes. AB - HYPOTHESIS: The use of passenger compartment safety measures has not led to decreases in pediatric morbidity or mortality in our population of patients. DESIGN: Retrospective review. SETTING: University, tertiary care, level I trauma center. PATIENTS: All patients admitted to the Trauma Center at Louisiana State University Health Science Center School of Medicine in Shreveport between July 1, 1991, and December 31, 2000, who were younger than 16 years and involved in a motor vehicle crash. MAIN OUTCOME MEASURES: Intensive care complications, postoperative complications, and mortality. RESULTS: We reviewed the experience of all pediatric patients involved in motor vehicle crashes and transported to the Trauma Center at Louisiana State University Health Science Center School of Medicine in Shreveport from July 1, 1991, through December 31, 2000. A total of 191 patients met these criteria. There were 8 deaths, and only 1 of these patients was restrained. There were significantly more injuries in those patients who died compared with those who survived (Modified Injury Severity Score, 29 vs 9; P<.001). We compared the use of restraints in our cohort with the use of restraints in the US pediatric population. Only 20% of our patients were restrained vs 68% of the general pediatric population. This difference was significant (P<.001, chi2) test). CONCLUSIONS: In our population of patients, death was a relatively infrequent occurrence. All patients who died presented in extremis. No patient died as the result of a complication. The rate of seat belt use in our population of patients was low. The exact reason for why we were unable to detect any survival benefit with seat belt use is unclear and demands further investigation. PMID- 12578408 TI - The value of color flow Doppler ultrasonography of the superior thyroid artery in the surgical management of Graves disease. AB - HYPOTHESIS: The factors affecting blood flow within the hypervascular thyroid gland and the effect of vascularization on the preparation for thyroidectomy as treatment for Graves disease can be documented. DESIGN: Blood flow through the superior thyroid arteries of patients with Graves disease, maintained in a euthyroid status, was measured by color flow Doppler ultrasonography. The microvessel density was assessed immunohistochemically using the level of expression of factor VIII in tissue sections. Both the thyroid gland's weight and blood loss volume were measured during the operation. SETTING: Tertiary care teaching hospital. PATIENTS: Fifty-two patients with Graves disease undergoing thyroidectomy. RESULTS: The blood flow rate was significantly correlated with thyroid weight (P<.01), thyroid microvessel density (P<.001), and histopathologic microscopic pattern (P<.001). The relation between these factors could be expressed as follows: Blood flow (mL/min) = [[0.0158 + 0.00136] x (weight (g) x microvessel density)](1/2) (R2 = 0.64, P<.001). Diffuse microfollicular hyperplastic thyroid tissue had a significantly higher blood flow and vascular density than tissue having an inactive colloid pattern (P<.01). Ten patients having a blood loss exceeding 200 mL during thyroidectomy showed a higher preoperative blood flow rate and microvessel density (P<.01). Of 8 patients whose blood flow was more than 0.15 L/min, 6 (75%) had blood loss in excess of 200 mL during surgery. CONCLUSIONS: The blood flow of the superior thyroid artery is positively related to intrathyroid microvessel density, glandular weight, and histopathologic microscopic pattern. Preoperative color flow Doppler ultrasonography may help in identifying patients with Graves disease who are liable to bleed intraoperatively during thyroidectomy. PMID- 12578410 TI - Differential diagnosis, investigation, and current treatment of lower limb lymphedema. AB - HYPOTHESIS: The causes and management of lower limb lymphedema in the Western population are different from those in the developing world. OBJECTIVE: To look at the differential diagnosis, methods of investigation, and available treatments for lower limb lymphedema in the West. DATA SOURCE: A PubMed search was conducted for the years 1980-2002 with the keyword "lymphedema." English language and human subject abstracts only were analyzed, and only those articles dealing with lower limb lymphedema were further reviewed. Other articles were extracted from cross referencing. RESULTS: Four hundred twenty-five review articles pertaining to lymphedema were initially examined. This review summarizes the findings of relevant articles along with our own practice regarding the management of lymphedema. CONCLUSIONS: The common differential diagnosis in Western patients with lower limb swelling is secondary lymphedema, venous disease, lipedema, and adverse reaction to ipsilateral limb surgery. Lymphedema can be confirmed by a lymphoscintigram, computed tomography, magnetic resonance imaging, or ultrasound. The lymphatic anatomy is demonstrated with lymphoscintigraphy, which is particularly indicated if surgical intervention is being considered. The treatment of choice for lymphedema is multidisciplinary. In the first instance, combined physical therapy should be commenced (complete decongestive therapy), with surgery reserved for a small number of cases. PMID- 12578411 TI - Duodenal-preserving resection of the head of the pancreas and pancreatic head resection with second-portion duodenectomy for benign lesions, low-grade malignancies, and early carcinoma involving the periampullary region. AB - HYPOTHESIS: Duodenal-preserving resection of the head of the pancreas (DPRHP) and pancreas head resection with segmental duodenectomy (PHRSD) can be alternatives to standard pancreaticoduodenectomy for benign periampullary lesions. DESIGN: Retrospective analysis of patients requiring surgery for benign and borderline malignant tumors of the periampullary region. SETTING: Tertiary care referral center. PATIENTS: Duodenal-preserving resection of the head of the pancreas (n = 8) and PHRSD (n = 7) were performed in 15 patients with a preoperative diagnosis of benign and borderline malignant tumors of the periampullary region (ie, 11 pancreas head lesions [2 intraductal papillary mucinous tumors, 4 serous cystadenomas, 2 insulinomas, 1 epidermal cyst, 1 metastatic renal cell carcinoma, 1 nonfunctioning islet cell tumor/parapaillary] and 4 duodenal lesions [3 adenomas and 1 adenocarcinoma]). MAIN OUTCOME MEASURES: Surgical factors (operation time and blood loss), postoperative complication, postoperative pancreatic insufficiency (eg, development of diabetes mellitus and steatorrhea or elevated stool elastase values), weight change, and recurrence of disease. RESULTS: No differences were noted in the mean operation time and estimated blood loss between the 2 procedures. Major postoperative complication constituted the following: bile duct stricture (n = 1) in DPRHP and delayed gastric emptying (n = 1) and postoperative bleeding (n = 1) in PHRSD. Newly developed diabetes mellitus occurred in 1 patient. Exocrine pancreatic insufficiency (steatorrhea) was observed in 1 patient after PHRSD. Patients with early duodenal carcinoma and intraductal papillary mucinous tumors with a borderline malignancy are still alive without evidence of recurrence. There was no hospital or long-term mortality. CONCLUSIONS: Duodenal-preserving resection of the head of the pancreas is recommended first for a benign or low-grade, early malignant pancreatic head lesion; PHRSD can be an option for a lesion of the ampullary-parapapillary duodenal area as well as the pancreatic head. Duodenal-preserving resection of the head of the pancreas can be converted to PHRSD if ischemia of the second portion of the duodenum occurs. We found benign periampullary lesions could be conservatively treated with DPRHP and PHRSD, which could substitute for classic pancreaticoduodenectomy. PMID- 12578413 TI - Influence of fever on the hypermetabolic response in burn-injured children. AB - BACKGROUND: Burn injury typically elicits a hypermetabolic response characterized by increased energy expenditure and muscle protein catabolism. HYPOTHESIS: Fever further increases energy expenditure and muscle loss in otherwise highly hypermetabolic burn patients. DESIGN: Retrospective analysis of experimental study. SETTING: University hospital. PATIENTS: Eighty-four children (aged 2-18 years) with burns covering 40% or more of total body surface area. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Simultaneous measurements of indirect calorimetry and leg net balance of phenylalanine (as an index of muscle protein catabolism) were obtained. Patients were stratified by their rectal temperature taken at the time of these metabolic measurements: afebrile (n = 28; temperature, <39.0 degrees C); mild fever (n = 26; temperature, 39.0 degrees C-39.4 degrees C); moderate fever (n = 18; temperature, 39.5 degrees C-39.9 degrees C); or severe fever (n = 12; temperature, > or =40.0 degrees C). RESULTS: Febrile and afebrile patients were similar in age, body weight, and extent of burn area. Severe fever was associated with significantly increased resting energy expenditure (mean +/- SD resting energy expenditure-predicted basal, 1.38 +/- 0.39 for afebrile patients vs 1.68 +/- 0.30 for patients with severe fever; P<.05) and a greater net loss of phenylalanine from the leg (net balance of phenylalanine, -6.0 +/- 6.2 mg/min per 100 mL of leg volume for afebrile patients vs -10.8 +/- 7.2 mg/min per 100 mL for patients with severe fever; P<.05). Patient groups were similar in plasma glucose concentration and extent of leukocytosis. CONCLUSIONS: These findings demonstrate the association of severe fever with further increase in energy expenditure and muscle protein catabolism in otherwise hypermetabolic burned children. This suggests a possible metabolic benefit in attenuating fever in such patients. PMID- 12578415 TI - Systemic temperature and paralysis after thoracoabdominal and descending aortic operations. AB - HYPOTHESIS: Systemic temperature influences the development of neurologic deficits after aortic surgery. DESIGN: Retrospective case-comparison study of prospectively collected data. SETTING: Tertiary referral center. PATIENTS AND INTERVENTIONS: We examined spinal cord injury according to mild passive hypothermia (mean temperature, 36.5 degrees C; n = 25), moderate active hypothermia (temperature range, 29 degrees C-32 degrees C; n = 76), or profound hypothermia (temperature, <20 degrees C; n = 31) for complex repairs in 132 patients. Aortic dissection was present in 67 patients (51%), 41 (31%) had leaks or rupture, 39 (30%) were reoperations on the descending thoracic aorta, and 27 (20%) had concurrent arch and/or ascending thoracic aortic repairs. MAIN OUTCOME MEASURE: Occurrence of permanent and transient deficits. RESULTS: Five patients (3.8%) had permanent deficits. One (4.0%) of the 25 patients underwent mild hypothermia, 3 (3.9%) of the 76 patients who underwent moderate hypothermia, and 1 (3.2%) of the 31 patients who underwent profound hypothermia (P =.70). Reversible deficits occurred in 7 patients (total 32%) who underwent mild hypothermia, 2 patients (total 6.6%) underwent moderate hypothermia, and 1 (total 6.5%) underwent profound hypothermia (P =.004). Six were delayed neurologic deficits. Independent predictors were intercostal ischemic time (P =.02), mild hypothermia (P =.004), and no cerebrospinal fluid drainage (P =.05). The total 30 day survival was 92.4% (122 of 132 patients). The only multivariable predictor of death was acuity of surgery (namely, emergent, urgent, or elective) (P =.06). CONCLUSIONS: Moderate or profound hypothermia resulted in fewer transient neurologic deficits. Thus, we recommend active cooling and cerebrospinal fluid drainage for most patients, and profound hypothermia for patients undergoing complex repairs and reoperations. PMID- 12578417 TI - Gastrojejunostomy during laparoscopic gastric bypass: analysis of 3 techniques. AB - HYPOTHESIS: Although perceived as a more technically demanding and time-consuming technique, the hand-sewn gastrojejunostomy during laparoscopic Roux-en-Y gastric bypass (RYGB) is associated with fewer complications and lower costs than stapled techniques. DESIGN: A retrospective medical record review of prospectively collected data. SETTING: University hospital. PATIENTS: One hundred eight consecutive patients undergoing laparoscopic RYGB between January 1, 1999, and December 31, 2001. INTERVENTION: Three techniques were compared: hand-sewn anastomosis (HSA), circular-stapled anastomosis (CSA), and linear-stapled anastomosis (LSA). MAIN OUTCOME MEASURES: Operative costs, including the cost of stapling devices, the cost of sutures, and operative times, were compared. Rates of anastomotic strictures, leaks, marginal ulcers, bleeding, and wound infections were determined. RESULTS: Eighty-seven patients underwent HSA; 13, CSA; and 8, LSA. Supply costs per patient were higher for CSA ($955) and LSA ($435) than for HSA ($2) (P<.001). The mean +/- SEM operative time for laparoscopic RYGB was longer when performing CSA than HSA or LSA (285 +/- 22 vs 215 +/- 8 and 204 +/- 28 minutes, respectively; P<.001). Stricture rates were higher after CSA than HSA and LSA (4 [31%] of 13 patients vs 3 [3%] of 87 patients and 0 of 8 patients, respectively; P<.01). The wound infection rate was higher after CSA than HSA and LSA (3 [23%] of 13 patients vs 1 [1%] of 87 patients and 0 of 8 patients, respectively; P<.001). There was no difference in anastomotic bleeding, and no anastomotic leaks occurred. CONCLUSIONS: In this experience, hand-sewn gastrojejunostomy during laparoscopic RYGB reduced operating room supply costs and was completed faster than stapled techniques. However, these differences may reflect the learning curve because these techniques were used early in our experience. Lower postoperative stricture and wound infection rates seem to be the primary benefits of the HSA technique. PMID- 12578418 TI - Hepatic resection in the United States: indications, outcomes, and hospital procedural volumes from a nationally representative database. AB - BACKGROUND: Hepatic resection has become common in the United States for both primary and secondary hepatic tumors. HYPOTHESIS: Variation in outcomes after hepatic resection is related to patient characteristics, the indication for operation, and hospital procedural volume. DESIGN: Observational study using a nationally representative database. PATIENTS: All patients in the Nationwide Inpatient Sample for 1996 and 1997 with a primary procedure code for hepatic resection (N = 2097). MAIN OUTCOME MEASURES: Outcomes included in-hospital mortality and length of stay. Risk-adjusted analyses were performed using hierarchical multivariate models. RESULTS: Overall mortality for the 2097 patients was 5.8%. The most common indications for hepatic resection were secondary metastases (52%), primary hepatic malignancy (16%), biliary tract malignancy (10%), and benign hepatic tumor (5%). High-volume hospitals had a mortality rate of 3.9% vs 7.6% at low-volume hospitals (P<.001). In the multivariate analysis adjusting for patient case-mix, high-volume hospitals had a 40% lower risk of in-hospital mortality compared with low-volume hospitals (odds ratio, 0.60; 95% confidence interval, 0.39-0.92; P =.02). Other predictors of mortality in the multivariate analysis included age older than 65 years, hepatic lobectomy (vs wedge resection), primary hepatic malignancy (vs metastases), and the severity of underlying liver disease. CONCLUSIONS: Hospital procedural volume is an important predictor of mortality after hepatic resection. Patients who require resection of primary and secondary liver tumors should be offered referral to a high-volume center. PMID- 12578419 TI - Revascularization of the internal carotid artery for isolated, stenotic, and symptomatic kinking. AB - HYPOTHESIS: The operation for isolated, stenotic, and symptomatic kinking of the internal carotid artery is safe and effective in preventing stroke and relieving the symptoms of cerebral ischemia. DESIGN: A consecutive sample clinical study with a mean follow-up of 44 months. SETTING: The surgical department of an academic tertiary care center and an affiliated secondary care center. PATIENTS: Fifty-four patients with a mean age of 67 years underwent 55 revascularizations of the internal carotid artery. The surgical procedures consisted of the following: shortening and reimplantation in the common carotid artery in 36 cases, bypass grafting in 15 cases, and transposition into the external carotid artery in 4 cases. MAIN OUTCOME MEASURES: Cumulative survival, primary patency, and stroke-free and neurologic symptom-free rates expressed by standard life table analysis. RESULTS: No patients died in the postoperative period. The postoperative stroke rate was 1.8%. The cumulative rates (SEs) at 5 years were as follows: survival, 70% (10.2%); primary patency, 89% (7.8%); overall stroke free, 92% (6.8%); ipsilateral stroke free, 96% (5.3%); neurologic symptom free, 90% (7.5%); and ipsilateral symptom free, 93% (6.5%). CONCLUSION: Revascularization of the internal carotid artery for the treatment of isolated, stenotic, and symptomatic kinking is safe and effective in preventing stroke and relieving symptoms of cerebrovascular insufficiency. PMID- 12578420 TI - Follow-up of infrainguinal graft thrombolysis: analysis of predictors of clinical success. AB - HYPOTHESIS: Thrombolysis is an accepted technique to salvage a failed infrainguinal bypass graft. Careful case selection, including consideration of the native arterial runoff and the type and location of the graft, will portend a better clinical outcome and prolonged graft patency. DESIGN: Retrospective study of an inception cohort of 91 acutely thrombosed grafts. SETTING: Academic tertiary care center. PATIENTS: We analyzed 91 consecutive occluded grafts in 69 patients for secondary graft patency and clinical outcome. INTERVENTION: Regional transcatheter thrombolysis. MAIN OUTCOME MEASURES: Technical success, secondary graft patency, and the need for major limb amputation. RESULTS: Immediate technical success resulting in restoration of flow was achieved in 80 (88%) of 91 cases. Angioplasty or additional surgical intervention (eg, patch, interposition graft, or jump graft to a more distal site) was performed in 44 subjects (64%). Longer duration of secondary patency was associated with synthetic vs vein grafts (P =.03), popliteal vs distal (tibial/pedal) insertion of the anastomosis (P =.008), and intact native arterial outflow (P =.003). Twenty-three cases required major limb amputation in the follow-up period, but 17 (74%) of these had reocclusion within 30 days of thrombolysis. Only 43 grafts (47%) were found to be patent at 1-year follow-up. CONCLUSIONS: In carefully selected cases, thrombolytic therapy is an effective means to restore limb viability in patients with occluded infrainguinal grafts. Long-term patency rates, although similar to those of surgical series, remain poor. PMID- 12578421 TI - Laparoscopic parastomal hernia repair. AB - Historically, parastomal hernias have represented a challenge to surgeons. Unsatisfying results from surgical repair have led to many modifications over time. Surgical repair of parastomal hernias has evolved from the traditional open technique to the current laparoscopic approach. We outline a technique that combines mesh reinforcement with a minimally invasive approach. PMID- 12578422 TI - Mechanisms and treatment of postoperative ileus. AB - OBJECTIVE: To review the pathogenesis and treatment of postoperative ileus. DATA SOURCES: Data collected for this review were identified from a MEDLINE database search of the English-language literature. The exact indexing terms were "postoperative ileus," "treatment," "etiology," and "pathophysiology." Previous review articles and pertinent references from those articles were also used. STUDY SELECTION: All relevant studies were included. Only articles that were case presentations or that mentioned postoperative ileus in passing were excluded. DATA SYNTHESIS: The pathogenesis of postoperative ileus is complex, with multiple factors contributing either simultaneously or at various times during the development of this entity. These factors include inhibitory effects of sympathetic input; release of hormones, neurotransmitters, and other mediators; an inflammatory reaction; and the effects of anesthetics and analgesics. Numerous treatments have been used to alleviate postoperative ileus without much success. CONCLUSIONS: The etiology of postoperative ileus can best be described as multifactorial. A multimodality treatment approach should include limiting the administration of agents known to contribute to postoperative ileus (narcotics), using thoracic epidurals with local anesthetics when possible, and selectively applying nasogastric decompression. PMID- 12578423 TI - Surgery in Lebanon. PMID- 12578424 TI - Diagnostic implications of C-reactive protein. AB - An elevated level of C-reactive protein (CRP), an acute phase protein, is one of many downstream indicators of inflammation. Physiologically, CRP enhances cell mediated immunity by promoting phagocytosis, accelerating chemotaxis, and activating platelets. The purposes of this article are (1) to review the clinical data implicating serum CRP level as a systemic marker of focal inflammation and (2) to explore serum CRP level as a reflection of the inflammatory component of atherogenesis. Our findings indicate that CRP levels serve as an early marker of the magnitude of inflammation in events as dissimilar as appendicitis and myocardial infarction. The level of circulating CRP correlates with endovascular disease and may serve to identify otherwise asymptomatic patients at sufficient cardiovascular risk to warrant aggressive therapy. Determining whether CRP has a direct pathologic role in the vascular wall itself may have the most clinical relevance. PMID- 12578425 TI - Image of the month. Internal herniation of the transverse colon. PMID- 12578426 TI - Crystalloid resuscitation: a waste of blood? PMID- 12578427 TI - A glass half full. PMID- 12578428 TI - Waterlilies: green reflections. PMID- 12578429 TI - Outpatient prescriptions for atypical antipsychotics for African Americans, Hispanics, and whites in the United States. AB - BACKGROUND: New antipsychotic medications introduced during the past decade clozapine (1990), risperidone (1994), olanzapine (1996), and quetiapine fumarate (1997)-offer a decrease in serious adverse effects compared with traditional antipsychotic medications, but at up to 10 times the cost. We examined whether ethnic minorities achieve access to these new advanced treatments. METHODS: Using national data on physician office and hospital outpatient department visits from the National Ambulatory Medical Care Survey and National Hospital Ambulatory Medical Care Survey from 1992 through 2000, we selected all patient visits at which an antipsychotic medication (atypical or traditional) was prescribed or continued and the patient was aged between 18 and 69 years. We performed a series of cross-sectional logistic regression analyses to determine the association of ethnic group and receipt of an atypical antipsychotic prescription over time, adjusted for potential confounders such as age, diagnosis, and health insurance type. RESULTS: Antipsychotic medication was prescribed or continued in 5032 visits; 33% of overall visits involved an atypical antipsychotic prescription. During 1992 to 1994, the adjusted relative odds of receipt of an atypical antipsychotic prescription for African Americans was 0.50 (95% confidence interval [CI], 0.26-0.96) and for Hispanics was 0.43 (95% CI, 0.16-1.18) compared with whites. During 1995 to 1997, the odds of receipt of a prescription for atypical antipsychotics increased for African Americans (odds ratio [OR], 0.69; 95% CI, 0.54-0.85) and for Hispanics (OR, 0.84; 95% CI, 0.65-1.07) compared with whites; and during 1998 to 2000, the relative odds continued to increase for African Americans (OR, 0.88; 95% CI, 0.78-0.97) and for Hispanics (OR, 1.05; 95% CI, 0.92-1.16) compared with whites. For visits specified for psychotic disorders, receipt of atypical antipsychotics was still lower for African Americans by 1998 to 2000 (adjusted OR, 0.74; 95% CI, 0.61-0.89) compared with whites, while for Hispanics the relative odds was equivalent (adjusted OR, 1.05; 95% CI, 0.87-1.19). CONCLUSION: Early gaps between ethnic groups in receipt of atypical antipsychotic prescriptions decreased throughout the 1990s but persisted for African Americans with psychotic disorders. PMID- 12578430 TI - Dehydroepiandrosterone augmentation in the management of negative, depressive, and anxiety symptoms in schizophrenia. AB - CONTEXT: Negative symptoms of schizophrenia are a prominent feature of the illness, and frequently remain refractory to treatment. Dehydroepiandrosterone (DHEA), along with its sulfated form, DHEA-S, is an important circulating neurosteroid with several vital neurophysiological functions, including the regulation of neuronal excitability and function. OBJECTIVE: Since the administration of DHEA has demonstrated improvement in mood, sense of well-being, interest, activity, and energy in several subpopulations, we investigate the efficacy of DHEA in the management of the negative symptoms of schizophrenia. DESIGN: Thirty DSM-IV-diagnosed schizophrenic patients with prominent negative symptoms (inpatients in a large referral state hospital) were randomized to receive either DHEA or placebo in double-blind fashion, in addition to regular antipsychotic medication, dose-stabilized prior to study entry. The DHEA was titrated up to a dose of 100 mg in divided doses during 6 weeks. RESULTS: Results indicated significant improvement in negative symptoms (P<.001), as well as in depressive (P<.05) and anxiety (P<.001) symptoms in individuals receiving DHEA. This effect was especially noted in women. The improvement in negative symptoms was independent of improvement in depression. No differences were noted on the positive symptom subscale of the Positive and Negative Syndrome Scale (PANSS) or on the total PANSS score as compared with placebo. Subjects receiving DHEA demonstrated a significant increase in DHEA (P<.05) and DHEA-S (P<.01) plasma levels, without changes in cortisol levels. Increases in DHEA and plasma DHEA-S levels were correlated with improvement in negative symptoms (P<.05), but not with improvement in depressive and anxiety symptoms. No obvious adverse effects were experienced by participating subjects. CONCLUSIONS: Our preliminary observations report for the first time in double-blind fashion the efficacy of DHEA augmentation in the management of negative, depressive, and anxiety symptoms of schizophrenia. The findings from this study raise important issues regarding the role of neurosteroids in general, and DHEA in particular, in the ongoing symptomatology and pharmacotherapy of schizophrenia. PMID- 12578431 TI - A randomized controlled study of cognitive therapy for relapse prevention for bipolar affective disorder: outcome of the first year. AB - BACKGROUND: Despite the use of mood stabilizers, a significant proportion of patients with bipolar affective disorder experience frequent relapses. A pilot study of cognitive therapy (CT) specifically designed to prevent relapses for bipolar affective disorder showed encouraging results when used in conjunction with mood stabilizers. This article reports the outcome of a randomized controlled study of CT to help prevent relapses and promote social functioning. METHODS: We randomized 103 patients with bipolar 1 disorder according to the DSM IV, who experienced frequent relapses despite the prescription of commonly used mood stabilizers, into a CT group or control group. Both the control and CT groups received mood stabilizers and regular psychiatric follow-up. In addition, the CT group received an average of 14 sessions of CT during the first 6 months and 2 booster sessions in the second 6 months. RESULTS: During the 12-month period, the CT group had significantly fewer bipolar episodes, days in a bipolar episode, and number of admissions for this type of episode. The CT group also had significantly higher social functioning. During these 12 months, the CT group showed less mood symptoms on the monthly mood questionnaires. Furthermore, there was significantly less fluctuation in manic symptoms in the CT group. The CT group also coped better with manic prodromes at 12 months. CONCLUSION: Our findings support the conclusion that CT specifically designed for relapse prevention in bipolar affective disorder is a useful tool in conjunction with mood stabilizers. PMID- 12578432 TI - Lifetime history of depression and carotid atherosclerosis in middle-aged women. AB - BACKGROUND: Depression is associated with clinical coronary events, but the association between history of major depression and subclinical cardiovascular disease in women is not yet known. We determined the association between lifetime history of major depression and subclinical carotid atherosclerosis in middle aged women. METHODS: Participants included 336 healthy middle-aged women (one third African American) from 1 of the 7 sites of the Study of Women's Health Around the Nation, a prospective study of the perimenopausal transition. Psychiatric diagnoses were assessed using the Structured Clinical Interview for the DSM-IV Axis I Disorders-Non-Patient Edition. Two measures of subclinical carotid atherosclerosis were assessed using B-mode ultrasonography: plaque and intima-media thickness. RESULTS: Lifetime history of major depression was associated with plaque, and substance abuse was related to intima-media thickness. Lifetime history of an anxiety disorder was not associated with either measure. After controlling for standard cardiovascular risk factors, only the association between major depression and plaque was maintained. The risk of plaque was 2-fold in women with a lifetime history of recurrent major depressive episodes relative to women with no history of depression (odds ratio = 2.30; 95% confidence interval, 1.10-4.82). Lifetime history of a single major depressive episode was not associated with plaque. CONCLUSIONS: Recurrent major depressive episodes may be a risk factor for subclinical atherosclerosis. Prevention of recurrent episodes may also prevent further progression of atherosclerosis. PMID- 12578433 TI - Induced panic attacks shift gamma-aminobutyric acid type A receptor modulatory neuroactive steroid composition in patients with panic disorder: preliminary results. AB - BACKGROUND: Certain metabolites of progesterone such as 3alpha,5alpha tetrahydroprogesterone (3alpha,5alpha-THP; allopregnanolone) and 3alpha,5beta-THP (pregnanolone) are potent, positive allosteric modulators of gamma-aminobutyric acid type A receptors. Although animal studies suggest anxiolytic properties of these endogenous modulators of central nervous excitability, no clinical data indicate whether they are also involved in the pathophysiology of anxiety disorders and panic attacks. METHODS: We quantified the concentrations of 3alpha,5alpha-THP, 3alpha,5beta-THP, the isomer 3beta,5alpha-THP, and their precursors in the plasma of 10 patients with panic disorder and matched control subjects during panic attacks induced by means of sodium lactate and cholecystokinin tetrapeptide administration, using a highly sensitive gas chromatography-mass spectrometry analysis. RESULTS: Panic attacks induced by sodium lactate and cholecystokinin tetrapeptide in patients with panic disorder were accompanied by pronounced decreases in the concentrations of 3alpha,5alpha THP and 3alpha,5beta-THP and a concomitant increase in the concentrations of the functional antagonistic isomer 3beta,5alpha-THP, findings that are compatible with a decreased gamma-aminobutyric acid-ergic tone. No changes in neuroactive steroid concentrations were observed after placebo administration in patients with panic disorder or after placebo, sodium lactate, or cholecystokinin tetrapeptide administration in controls. CONCLUSIONS: The association between changes in plasma neuroactive steroid concentrations and experimentally induced panic attacks and the well-documented pharmacological properties of these compounds as gamma-aminobutyric acid type A receptor modulators suggest that neuroactive steroids may play a role in the pathophysiology of panic attacks in patients with panic disorder. PMID- 12578434 TI - Family study of affective spectrum disorder. AB - BACKGROUND: Affective spectrum disorder (ASD) represents a group of psychiatric and medical conditions, each known to respond to several chemical families of antidepressant medications and hence possibly linked by common heritable abnormalities. Forms of ASD include major depressive disorder (MDD), attention deficit/hyperactivity disorder, bulimia nervosa, cataplexy, dysthymic disorder, fibromyalgia, generalized anxiety disorder, irritable bowel syndrome, migraine, obsessive-compulsive disorder, panic disorder, posttraumatic stress disorder, premenstrual dysphoric disorder, and social phobia. Two predictions of the ASD hypothesis were tested: that ASD, taken as a single entity, would aggregate in families and that MDD would coaggregate with other forms of ASD in families. METHODS: Probands with and without MDD, together with their first-degree relatives, were interviewed using the Structured Clinical Interview for DSM-IV and a supplemental interview for other forms of ASD. The familial aggregation and coaggregation of disorders were analyzed using proband predictive logistic regression models, including a novel bivariate model for the presence or absence of each of 2 disorders in a relative as predicted by the presence or absence of each of 2 disorders in the associated proband. RESULTS: In the 178 interviewed relatives of 64 probands with MDD and 152 relatives of 58 probands without MDD, the estimated odds ratio (95% confidence interval) for the familial aggregation of ASD as a whole was 2.5 (1.4-4.3; P =.001) and for the familial coaggregation of MDD with at least one other form of ASD was 1.9 (1.1-3.2; P =.02). CONCLUSIONS: Affective spectrum disorder aggregates strongly in families, and MDD displays a significant familial coaggregation with other forms of ASD, taken collectively. These results suggest that forms of ASD may share heritable pathophysiologic features. PMID- 12578435 TI - Predictors of mortality in eating disorders. AB - BACKGROUND: Anorexia nervosa, but not bulimia nervosa, has one of the highest mortality rates among psychiatric disorders. However, potential predictors of mortality, such as comorbid psychiatric illnesses, remain unclear. We sought to determine mortality ratios and predictors of fatal outcome in women diagnosed as having anorexia or bulimia nervosa. METHODS: Women (N = 246) diagnosed as having either DSM-IV anorexia nervosa (n = 136) or bulimia nervosa (n = 110) between January 1, 1987, and December 31, 1991, participated in a prospective longitudinal study. Vital status was determined by ongoing contact and a National Death Index search as of December 1998 (overall ascertainment, 99.8%) and telephone contact as of October 2000 (ascertainment, 95.0%). RESULTS: Eleven women died. Standardized mortality ratios were elevated for all causes of mortality (11.6; 95% confidence interval, 5.5-21.3) and suicide (56.9; 95% confidence interval, 15.3-145.7) in anorexia nervosa but not for death (1.3; 95% confidence interval, 0.0-7.2) in bulimia nervosa. Predictors of mortality in anorexia nervosa included severity of alcohol use disorder during follow-up (P<.001). Hospitalization for an affective disorder before baseline assessment seemed to protect women from a fatal outcome (P<.001). CONCLUSIONS: Physicians treating patients with anorexia nervosa should carefully assess patterns of alcohol use during the course of care because one third of women who had alcoholism and died had no history of alcohol use disorder at intake. PMID- 12578436 TI - Screening for serious mental illness in the general population. AB - BACKGROUND: Public Law 102-321 established a block grant for adults with "serious mental illness" (SMI) and required the Substance Abuse and Mental Health Services Administration (SAMHSA) to develop a method to estimate the prevalence of SMI. METHODS: Three SMI screening scales were developed for possible use in the SAMHSA National Household Survey on Drug Abuse: the Composite International Diagnostic Interview Short-Form (CIDI-SF) scale, the K10/K6 nonspecific distress scales, and the World Health Organization Disability Assessment Schedule (WHO-DAS). An enriched convenience sample of 155 respondents was administered all screening scales followed by the 12-month Structured Clinical Interview for DSM-IV and the Global Assessment of Functioning (GAF). We defined SMI as any 12-month DSM-IV disorder, other than a substance use disorder, with a GAF score of less than 60. RESULTS: All screening scales were significantly related to SMI. However, neither the CIDI-SF nor the WHO-DAS improved prediction significantly over the K10 or K6 scales. The area under the receiver operating characteristic curve of SMI was 0.854 for K10 and 0.865 for K6. The most efficient screening scale, K6, had a sensitivity (SE) of 0.36 (0.08) and a specificity of 0.96 (0.02) in predicting SMI. CONCLUSIONS: The brevity and accuracy of the K6 and K10 scales make them attractive screens for SMI. Routine inclusion of either scale in clinical studies would create an important, and heretofore missing, crosswalk between community and clinical epidemiology. PMID- 12578437 TI - Familial patterns of risk in very late-onset Alzheimer disease. AB - BACKGROUND: The incidence of Alzheimer disease (AD) peaks after 85 years of age. Although genetic factors are implicated in AD with substantially earlier onset, the familial characteristics of high-incidence very late-onset AD (VLOAD, defined here as AD with onset age >/=85 years) remain unknown. METHODS: We collected information pertaining to the cognitive status and demographics of 809 parents and siblings of 144 VLOAD probands, 4235 parents and siblings of 793 earlier onset AD probands, and 7646 parents and siblings of 1493 nondemented elderly probands. Cumulative risks and 5-year interval-specific hazard rate ratios were calculated for AD in relatives of the 2 AD proband groups and relatives of the nondemented elderly group. RESULTS: The cumulative risk for AD in the relatives of VLOAD probands was significantly different than that in the relatives of earlier-onset AD probands (P<.001), but not in the relatives of nondemented elderly probands. Also, the relatives of earlier-onset AD probands had hazard rate ratios ranging from 19.7 in those aged 50 to 54 years to 1.2 in those aged 90 to 94 years. Rates successively dropped as age intervals increased. CONCLUSIONS: At least through the middle of the ninth decade of life, relatives of VLOAD probands have a lower risk for AD than those of earlier-onset AD probands. In addition, the relatively increased risk of relatives of earlier onset AD probands is highest at younger ages and diminishes with increasing age. In counseling family members of patients with AD concerned about their own risk, the onset age of the patient and the age of the concerned relative should be considered. Very late-onset AD may be a good target for investigating environmental factors associated with AD. PMID- 12578438 TI - Serum anticholinergic activity in a community-based sample of older adults: relationship with cognitive performance. AB - BACKGROUND: Serum anticholinergic activity (SAA), as measured by a radioreceptor assay, quantifies a person's overall anticholinergic burden caused by all drugs and their metabolites. In several small geriatric patient groups, SAA has been associated with cognitive impairment or frank delirium. To our knowledge, there has not yet been any systematic study of the prevalence of SAA and its effect on cognition in a community-based population. METHODS: Serum anticholinergic activity was measured in 201 subjects who were randomly selected among the participants in an epidemiological community study, based on their age and sex. Cognitive performance was assessed with use of the Mini-Mental State Examination. The association between SAA and cognitive performance was examined using a univariate analysis and a multiple logistic regression model, adjusting for age, sex, educational level, and number of medications. RESULTS: Serum anticholinergic activity was detectable in 180 (89.6%) participants (range, 0.50-5.70 pmol/mL). Univariate testing showed a significant association between SAA and Mini-Mental State Examination scores. Logistic regression analysis indicated that subjects with SAA at or above the sample's 90th percentile (ie, SAA >/=2.80 pmol/mL) were 13 times (odds ratio, 1.08-152.39) more likely than subjects with undetectable SAA to have a Mini-Mental State Examination score of 24 (the sample's 10th percentile) or below. CONCLUSIONS: To our knowledge, this is the largest analysis of SAA and the first to examine its extent and relationship with cognitive performance in a community sample. Its results suggest that SAA can be detected in most older persons in the community and confirm that even low SAA is associated with cognitive impairment. PMID- 12578440 TI - Dilating the clinical research stricture. PMID- 12578442 TI - A gift to heal. PMID- 12578439 TI - Development of a new once-a-day formulation of methylphenidate for the treatment of attention-deficit/hyperactivity disorder: proof-of-concept and proof-of product studies. AB - BACKGROUND: The duration of action of the immediate-release formulation of methylphenidate hydrochloride is short (3 to 4 hours), and 3 times daily dosing is thought to maximize effectiveness across a 12-hour day. The initial sustained release formulations of methylphenidate had reduced efficacy compared with immediate-release methylphenidate and were not well accepted. Tachyphylaxis was hypothesized to account for the reduced effects, and an ascending drug delivery pattern was proposed to overcome this acute tolerance. METHODS: Children with attention-deficit/hyperactivity disorder were evaluated in a laboratory school to characterize onset and duration of the effect of a variety of methylphenidate regimens. In a proof-of-concept study, an experimental ascending profile was established by an initial bolus followed by small increasing doses of immediate release methylphenidate in capsules administered every 30 minutes for 8 hours. Two proof-of-product studies of a new oral once-a-day formulation to deliver methylphenidate by an osmotic pump process based on OROS (ALZA Corp, Mountain View, Calif) technology (hereafter referred to "OROS-methylphenidate") were conducted: a pharmacokinetic study and a pharmacodynamic study. RESULTS: The experimental ascending profile matched the effect of the standard regimen of methylphenidate, 3 times daily. In the pharmacokinetic study, OROS methylphenidate treatment produced a rapid rise followed by increasing plasma concentrations that peaked 7 to 9 hours after administration. In the pharmacodynamic study, OROS-methylphenidate treatment matched the 3 times daily dosing of methylphenidate for onset and duration of efficacy. CONCLUSIONS: These studies demonstrate the translation of a basic science finding (acute tolerance to clinical doses of methylphenidate) into clinical application (the selection of a new drug delivery pattern for methylphenidate). This approach produced a new product (OROS-methylphenidate or Concerta), which proved to have the predicted rapid onset (with 1-2 hours) and long duration of efficacy (10-12 hours) after a single administration in the morning. PMID- 12578444 TI - Complications and determinants of length of stay for inpatient pediatric otolaryngologic procedures. AB - OBJECTIVE: To identify clinical factors associated with morbidity, mortality, and length of stay (LOS) for inpatient pediatric otolaryngologic procedures. STUDY DESIGN: Retrospective cohort study. METHODS: Records of patients undergoing pediatric otolaryngologic procedures were extracted from the National Hospital Data Survey for the calendar years 1995 through 1999. These records were examined to determine demographics, morbidity, mortality, type of procedure (as defined by anatomic subsite), and LOS. The effects of morbidity and type of procedure on LOS were identified. RESULTS: A total of 4861 children underwent inpatient otolaryngologic procedures. The overall morbidity rate was 4.6%. The most common morbidity was pneumonia, occurring in 171 patients (3.5%). The presence of any morbidity was associated with a significantly increased LOS (18.4 days vs 4.6 days; P<.001). The occurrence of pneumonia was associated with an increase in LOS to 19.7 days vs 4.7 days in patients without pneumonia (P<.001). Procedures involving the larynx, trachea, or esophagus carried the longest LOS (12.3 days; P<.001) among all procedural categories. The overall mortality rate was distinctly low at 0.4%. However, the occurrence of morbidity was associated with an increased risk of death, with an odds ratio of 8.0 (P =.001). Mortality was highest (13 of 18 deaths) after procedures on the larynx, trachea, or esophagus. CONCLUSIONS: Medical complications in children undergoing inpatient otolaryngologic procedures are associated with significantly increased LOS. Despite procedural complexity, overall mortality is remarkably low. Efforts to decrease medical morbidity in this population may result in decreased LOS and improved clinical outcomes. PMID- 12578443 TI - Prevention of otitis media by adenoidectomy in children younger than 2 years. AB - OBJECTIVES: To test the effect of adenoidectomy in connection with tympanostomy compared with tympanostomy only in preventing otitis media in children younger than 2 years. DESIGN: Prospective trial with randomized and nonrandomized arms. SETTING: Primary care study clinics. PARTICIPANTS AND INTERVENTIONS: The study participants were selected from 2497 children who had been enrolled in the Finnish Otitis Media Vaccine Trial at the age of 2 months. A total of 306 children, aged 1 to 2 years, who had experienced recurrent episodes of otitis media were randomized into 2 treatment groups: tympanostomy with or without adenoidectomy. Of the 306 children, 137 were operated on according to random basis (randomized trial). The 169 children whose parents declined participation in the randomized trial were operated on according to the parents' preferences (nonrandomized trial). All children were followed up until 2 years of age. The mean follow-up time was 7 months. MAIN OUTCOME MEASURE: The rate of acute otitis media episodes. RESULTS: The average reduction in the rate of all acute otitis media episodes in the adenoidectomy group was 19% (95% confidence interval [CI], 14% to 43%) among children enrolled in the randomized trial and 25% (95% CI, -13% to 50%) in the nonrandomized trial. The reduction in the randomized trial was mainly due to reduction in the rate of pneumococcal otitis media (58%, 95% CI, 16%-79%). CONCLUSION: In children younger than 2 years, concurrent adenoidectomy during the insertion of tympanostomy tubes does not seem to have a major advantage over the insertion of tympanostomy tubes alone in preventing otitis media. PMID- 12578445 TI - Relation of age to outcome after endoscopic sinus surgery in children. AB - OBJECTIVE: To determine whether endoscopic sinus surgery (ESS) in children has a better outcome at a certain age. DESIGN AND SETTING: Cohort study in a tertiary care children's hospital. PATIENTS: Ninety-nine children who underwent ESS between January 1994 and June 1999. MAIN OUTCOME MEASURES: Determining ESS outcomes was based on answers to a questionnaire mailed to the caregivers at least 1 year after surgery. Outcomes were considered failures if children required revision surgery or if their symptoms were not improved. RESULTS: The overall ESS success rate was 82%. Univariate analysis of age and surgery outcome revealed that children older than 6 years had an 89% success rate but that children younger than 6 years had a 73% success rate (P =.04). Of the 99 patients, 11 (9%) required revision surgery. Among them, 9 were younger and 2 were older than 6 years (P =.008). Of the 4 children younger than 3 years, 3 (75%) required revision surgery. CONCLUSIONS: Endoscopic sinus surgery in children younger than 3 years was not successful but it was beneficial in children older than 6 years. It may be beneficial at any age if a complication from chronic sinusitis occurs; however, for children younger than 6 years, revision surgery may later be necessary. PMID- 12578446 TI - Outcomes in suction coagulator adenoidectomy. AB - OBJECTIVES: To assess the feasibility of suction coagulator adenoidectomy relative to adenoid size and to document patient-based outcomes and satisfaction with surgery. DESIGN: Historical cohort study. SETTING: Referral-based academic pediatric otolaryngology practice. PARTICIPANTS: Consecutive series of 118 children older than 3 years (mean age, 6.5 years) undergoing adenoidectomy alone. INTERVENTION: Video nasopharyngoscopy followed by suction coagulator adenoidectomy as part of routine clinical care. An outcome survey was completed by telephone. OUTCOME MEASURES: Duration of surgery, estimated blood loss, complications, parent satisfaction, and clinical outcomes. RESULTS: The distribution of preoperative adenoid grades as determined by nasal endoscopy were as follows: grade 2 (>or=33% to <66% choanal obstruction), 7%; grade 3 (>or=66% to <90% obstruction), 48%; and grade 4 (90%-100% obstruction), 45%. The mean (SD) surgical time of 10.5 (3.0) minutes was unrelated to adenoid grade (R = 0.014; P =.88). All blood loss was less than 15 mL (<5 mL for 67%), and the only complication was a loose tooth. The parents of 98 patients (83%) were contacted a mean of 30.4 days after surgery: 95 (97%) reported less-labored breathing, and 94 (96%) were satisfied with the surgical results. Only 5 patients (5%) required a follow-up visit within 30 days of surgery. CONCLUSIONS: Suction coagulator adenoidectomy proved safe and rapid, regardless of the adenoid size. Surgical outcomes were very favorable, with nearly all parents reporting satisfaction with the procedure and improvement in their child's breathing. PMID- 12578447 TI - Use of the operating microscope and loupes for head and neck free microvascular tissue transfer: a retrospective comparison. AB - OBJECTIVE: To analyze whether less cumbersome loupes are as equally efficacious as the microscope for magnification for free tissue transfers in head and neck reconstruction. DESIGN: A 7-year retrospective review of 151 consecutive microvascular free tissue transfers compares the operating microscope with loupe magnification. SETTING: A tertiary care medical center. PATIENTS: A total of 151 consecutive patients undergoing microvascular reconstructions. INTERVENTION: Microvascular reconstruction of ablative head and neck cancer defects using either microscopic or loupe-provided magnification. MAIN OUTCOME MEASURES: Medical records were reviewed for demographic data, preoperative radiation status, type of defect and flap, length of procedure, complications, and length of stay. RESULTS: Two teams shared the reconstructions, with all microvascular anastomoses created or directly supervised by one of us (D.A.R.). One team used the operating microscope for magnification (84 cases), while the other used loupes (67 cases). Complication rates were statistically similar for the 2 techniques, the length of procedure tended to be shorter in the loupe group, and the length of stay was statistically similar in both groups. CONCLUSIONS: A review of the literature validates the usefulness of loupe magnification in free tissue transfer surgery. To our knowledge, this is the largest number of head and neck reconstruction cases presented comparing these techniques. Loupes offer the surgeon comfort and easy access to and uncomplicated visualization of the operating field. Furthermore, loupes may decrease operating time and help avoid complicated equipment. Therefore, loupe magnification should be considered in head and neck reconstruction without the fear of increased morbidity. PMID- 12578448 TI - Possible impedance of luminal reepithelialization by tracheal cartilage metalloproteinases. AB - BACKGROUND: Rapid reepithelialization of respiratory epithelium after injury to the large conducting airway (eg, trachea and bronchus) is poor. Our laboratory has developed an in vitro model of the trachea that allows us to examine reepithelialization in a complex culture system. We previously described how the presence of cartilage inhibited respiratory epithelial cell (REC) migration/proliferation. In the present study, we examined the effect of cartilage-conditioned medium (CCM) on REC proliferation. We hypothesized that a potential cause of delayed reepithelialization of the large conducting airway after injury could be excessive or aberrant secretion of matrix metalloproteinases (MMPs) by cartilage. DESIGN: We assessed cartilage-derived MMP production and effects on REC proliferation by adding CCM to primary cultures of porcine RECs on type I collagen and determining the cell number and viability. Cartilage-conditioned medium-derived MMP activity was determined by means of gelatin zymography in pooled samples from different times during in vitro cartilage culture. RESULTS: We detected MMP-2 and a small amount of MMP-9 in CCM. Enzyme activity was abolished by EDTA, confirming MMP identity. Cartilage conditioned medium inhibited REC attachment and proliferation. Addition of the MMP inhibitor GM6001 to cartilage cultures yielded CCM that did not inhibit REC growth, indicating a role for cartilage-derived MMPs in modulating REC proliferation. CONCLUSION: Cartilage production and activity of MMP after injury to the large conducting airway may be a factor in the failure of luminal reepithelialization, resulting in aberrant repair. PMID- 12578449 TI - A tissue-engineering technique for vascularized laryngotracheal reconstruction. AB - OBJECTIVE: To perform laryngotracheal reconstruction (LTR) using a vascularized neotracheal segment. DESIGN: A neotracheal segment was created within the sternocleidomastoid muscle. An anterior cricoid split procedure was performed using a pedicled, vascularized neotracheal segment. Results were compared with a control group that underwent anterior cricoid split using standard (avascular) autografted cartilage. Cross-sectional area, cartilage viability, extrusion, mucosalization, and wound healing were compared between groups. SUBJECTS: Sixteen female New Zealand white rabbits. INTERVENTIONS: Eight animals underwent placement of a cartilage-wrapped silicone implant into the sternocleidomastoid muscle. After 2 weeks, the silicone implant was removed, leaving a fibrovascular "foreign body" capsule and the interwoven autografted cartilage. The neotracheal segment was trimmed to create an anterior graft for LTR. The remaining animals underwent standard anterior graft LTR using autografted auricular cartilage. The reconstructed segments were harvested for comparison at 2 and 4 weeks. RESULTS: All reconstructed animals survived the postoperative period. No significant differences in stenosis rates or mucosalization were noted between groups. Two animals in the standard LTR group had microabscess formation, and no graft extrusions were encountered. CONCLUSION: A pedicled neotracheal graft can be used for anterior cricoid split procedures in rabbits. PMID- 12578450 TI - Survival and prognosis in Hurthle cell carcinoma of the thyroid gland. AB - OBJECTIVE: To determine factors that affect survival in patients with Hurthle cell carcinoma of the thyroid gland. METHODS: Data for all cases of Hurthle cell carcinoma that occurred between January 1, 1988, and December 31, 1998, were extracted from the Surveillance, Epidemiology, and End Results database. Clinical data regarding age, sex, tumor size, primary site extension, nodal involvement, and vital status were tabulated. Patients with distant metastases were excluded, and Kaplan-Meier survival analysis was conducted. Survival data for patients with Hurthle cell carcinoma were compared with data for a control group of patients with follicular cell carcinoma matched for age, sex, tumor size, and local disease extension. Cox multivariate regression analysis was conducted to determine the effect of predictor variables on overall survival in Hurthle cell carcinoma. RESULTS: We identified 555 cases of nonmetastatic Hurthle cell carcinoma (mean age at diagnosis, 55.9 years; women, 67.9%). The primary tumor was intrathyroidal in 83.8% of patients, whereas 11.2% had minor local extension. Mean tumor size was 3.5 cm. Mean, 5-year, and 10-year survival for Hurthle cell carcinoma was 109 months, 85.1%, and 71.1%, respectively. Mean survival for 411 matched patients with follicular cell carcinoma was 113 months, which was not statistically different from that of patients with Hurthle cell carcinoma (P =.47, log-rank test). On multivariate analysis, increasing age at diagnosis, male sex, and increasing tumor size were statistically significant predictors of poor survival; degree of primary site extension did not affect survival. CONCLUSIONS: Overall survival for Hurthle cell carcinoma is similar to that of comparably staged follicular cell carcinoma. Increasing age, male sex, and increasing tumor size substantially diminish survival in patients with Hurthle cell carcinoma. PMID- 12578451 TI - Expression of cathelicidin in human salivary glands. AB - BACKGROUND: Salivary secretions play a critical role in maintaining oral health via innate host defense mechanisms and secretion of secretory IgA. One of the antimicrobial peptides, LL-37, is the only cathelicidin protein that has yet been identified in humans. Cathelicidins are a family of peptides thought to provide an innate defensive barrier against a variety of potential microbial pathogens. OBJECTIVES: To examine the expression of cathelicidin in human salivary glands and to investigate up-regulation of cathelicidin in inflammatory conditions. DESIGN: Reverse transcriptase-polymerase chain reaction and immunohistochemical staining were performed on 20 salivary gland tissues, 10 from normal salivary glands and 10 from glands with chronic sialadenitis. RESULTS: Cathelicidin messenger RNA transcripts were detected in the tissues from the normal salivary glands and the glands with chronic sialadenitis. The level of cathelicidin messenger RNA in glands with chronic sialadenitis was significantly increased compared with that in normal salivary glands. Cathelicidin protein was expressed in the glandular epithelium of the normal salivary glands and the glands with chronic sialadenitis. CONCLUSION: The results indicate that cathelicidin might play an important role in innate host defense of human salivary glands. PMID- 12578453 TI - Perioperative complications, comorbidities, and survival in oral or oropharyngeal cancer. AB - OBJECTIVES: To establish the value of clinical factors in the prediction of perioperative complications and death in patients with oral and oropharyngeal carcinomas and to develop a new extended clinical severity staging system that combines patient and tumor factors. PATIENTS AND METHODS: A total of 530 patients with oral or oropharyngeal carcinomas submitted to surgical treatment were studied. Logistic regression was used to identify risk factors for perioperative complications, and the Cox proportional hazards regression model was used to establish independent prognostic factors. RESULTS: Daily alcohol consumption, smoking, sex, neck lump, earache, pain, dysphagia, weight loss, oral bleeding, odynophagia, body mass index, National Cancer Institute comorbidity index score, American Society of Anesthesiologists surgical risk, hematocrit level, and total lymphocyte count had an impact on prognosis in univariate analysis. Survival according to extended clinical severity stage was 76.7% for stage 1, 64.4% for stage 2, 44.8% for stage 3, and 25.5% for stage 4 (chi( 2) = 64.16; P<.001). In multivariate analysis, only APACHE II score, neck dissection, POSSUM index score, and type of reconstruction were independent risk factors for perioperative complications. The final prognostic model included development of local plus systemic complications, extended clinical severity stage, type of reconstruction, and APACHE II score. CONCLUSIONS: Clinical variables have a predictive effect on morbidity and mortality of patients with oral and oropharyngeal cancer treated surgically. Local plus systemic perioperative complications can adversely affect the prognosis. The uniformity of results confirms that survival estimates can be enhanced by the addition of clinical characteristics to the TNM classification, creating a more accurate system for the estimation of prognosis. PMID- 12578452 TI - Urokinase-type plasminogen activator receptor expression in adenoid cystic carcinoma of the skull base. AB - BACKGROUND: Head and neck adenoid cystic carcinoma (ACC) is a malignancy of the salivary and lacrimal glands with a variable growth pattern and propensity for perineural spread. Involvement of the skull base indicates a poor prognosis. Despite surgical resection and adjuvant radiotherapy, tumor recurrence and metastases are common. The urokinase-type plasminogen activator and its receptor (uPAR) have an important role in tumor invasion and metastasis. The expression of uPAR is predictive of poor outcomes in many tumors. This study examines the expression of human uPAR in ACCs involving the skull base. OBJECTIVES: To determine uPAR expression in ACCs of the skull base by immunohistochemical analysis and compare expression with tumor histologic findings and clinical outcomes. STUDY DESIGN: Analysis of uPAR in archival ACC specimens and a retrospective medical chart review. SETTING: Multidisciplinary cranial base program at a university medical center with tertiary referral pattern. RESULTS: Ten (83%) of 12 tumors stained positive for uPAR. Three of 3 patients who died of ACC and 6 of 6 patients alive with disease expressed uPAR. Only 1 of 3 patients free of disease was uPAR positive. CONCLUSIONS: In most patients with ACC of the skull base, uPAR was expressed. Its expression seems to be a negative prognostic factor. However, the small study sample limits our observations. Additional study of uPAR expression in ACC at other anatomic sites is required. PMID- 12578454 TI - Distension technique to improve computed tomographic evaluation of oral cavity lesions. AB - BACKGROUND: The apposing mucosa of the oral cavity makes the computed tomographic identification of a clinically obvious mass difficult. Contrast distension techniques have been used in radiology to evaluate for presence of a "hidden" mass. OBJECTIVE: To illustrate the utility of distending the oral cavity with air, water, or contrast to display otherwise obscure oral cavity lesions. PATIENTS AND METHODS: In 3 normal subjects and 5 patients with biopsy-proved oral cavity lesions, serial contiguous 3-mm axial and coronal computed tomographic scans were obtained before and after distension of the oral cavity using intraoral air or water. Air distension was achieved by having the subjects perform a modified Valsalva maneuver during the scan acquisitions. Fluid distension was obtained using approximately 40 mL of water. RESULTS: In each case, the contrast successfully distended the oral cavity, separating the mucosal surfaces. Gingivobuccal lesions that were obscured by apposition of the lips and cheeks to the gums and teeth, or by apposition of the tongue to the inner margins of the gums and teeth, were clearly demonstrated. Lesions involving or extending into the retromolar trigone were also well demonstrated using this distension technique. CONCLUSIONS: Computed tomographic display of the anatomy and pathology of the oral cavity can be improved simply by distending the oral cavity using air or water as a contrast medium. This technique successfully shows lesions that are obscured by the apposing surfaces of the vestibule and the oral cavity proper, improving computed tomographic diagnosis. PMID- 12578455 TI - Intraoperative fluorouracil in endonasal laser dacryocystorhinostomy. AB - BACKGROUND: Although endonasal laser dacryocystorhinostomy (ELDCR) offers many advantages compared with conventional techniques, postoperative scarring leading to occlusion of the rhinostomy is more common with ELDCR. OBJECTIVE: To investigate whether fluorouracil applied to the rhinostomy site intraoperatively has an effect on the outcome. PARTICIPANTS: We randomly allocated 155 consecutive patients (201 procedures) to a control group and a fluorouracil group. Patients and investigators were masked to the choice of treatment. METHODS: All patients underwent ELDCR. A pledget soaked in isotonic sodium chloride solution or 0.5 mg/mL fluorouracil, randomly allocated by the pharmacy, was applied to the rhinostomy site for 5 minutes at the time of surgery. Patients were followed up for 12 months and their symptoms were assessed at each visit. RESULTS: Among patients followed up for 12 months or longer, ELDCR procedures performed with topical application of fluorouracil to the rhinostomy site were successful in 65 (76%), compared with 52 (63%) for the control group. This was not statistically significant when patients who failed to attend follow-up at or after 12 months were not counted as successfully treated (P =.21, chi(2) test). Even if those who failed to attend for follow-up were counted as successes, the effect of fluorouracil did not reach significance at the.05 level (P =.08, chi(2) test). CONCLUSION: The topical application of fluorouracil failed to increase the patency rates in ELDCR. PMID- 12578456 TI - Computed tomographic findings in patients with invasive fungal sinusitis. AB - OBJECTIVE: To determine the radiographic findings of computed tomographic (CT) imaging most suggestive of invasive fungal sinusitis (IFS) in an immunocompromised patient population. DESIGN: A retrospective review of patients with a diagnosis of IFS reached with CT and confirmed by histopathologic evaluation. SETTING: An academic tertiary care hospital. PATIENTS: Twenty-three immunocompromised patients with confirmed IFS and preoperative CT imaging. Controls were 10 patients with acute myelocytic leukemia and CT evidence of sinusitis but no history of IFS. OUTCOME MEASURES: The CT scans were reviewed to identify factors predictive of invasive fungal disease. Parameters evaluated were nasal cavity and sinus soft tissue thickening, the presence of air-fluid levels, bone erosion, extrasinus extension, and unilateral or bilateral nasal cavity and sinus involvement. RESULTS: The CT findings included severe soft tissue edema of the nasal cavity mucosa (turbinates, lateral nasal wall and floor, and septum) in 21 of the 23 patients, sinus mucoperiosteal thickening in 21, bone erosion in 8, orbital invasion in 6, facial soft tissue swelling in 5, and retroantral fat pad thickening in 2. Two patients had air-fluid levels. No patients had intracranial involvement. Unilateral involvement was found in 21 patients, and bilateral involvement in 2. Review of the control group revealed only mild soft tissue edema of the nasal cavity in 2 (P<.001), unilateral involvement in 2 (P<.001), and evidence of bone erosion or extrasinus soft tissue involvement in none. CONCLUSIONS: Most patients do not have classic CT findings of bone erosion or extrasinus extension in the early course of IFS. We found that severe unilateral thickening of the nasal cavity mucosa was the most consistent finding on CT suggestive of underlying IFS, occurring much more frequently in immunocompromised patients with IFS than without IFS. Even though severe nasal cavity soft tissue thickening is much more common in IFS, this is a nonspecific finding that can be seen, to a lesser degree, in all forms of rhinosinusitis. Therefore, the clinician cannot rely solely on CT imaging and must maintain a high index of suspicion when evaluating immunocompromised patients to establish a prompt diagnosis. Early nasal endoscopy with biopsy and initiation of appropriate therapy are necessary to improve prognosis. PMID- 12578457 TI - Psammomatous calcification in association with a benign thyroglossal duct cyst. AB - Embryologic remnants of thyroid tissue often line the thyroglossal duct tract and may commonly become cystic. Calcification in such a cyst is thought to be a specific marker for malignancy, which may develop in 1% of thyroglossal duct cysts. We describe a 3-year-old boy with a midline neck mass that showed radiologic calcification. Pathologic evaluation revealed psammomatous calcification in association with a benign thyroglossal duct cyst. These findings, not previously reported (to our knowledge), expand the radiologic differential diagnosis of calcified neck masses and broaden the spectrum of pathologic findings seen in association with benign thyroglossal duct cysts. PMID- 12578458 TI - Chondroid chordoma of the nasal septum. AB - Chondroid chordoma is an uncommon malignant tumor of bone that occurs mostly in patients of early middle age. To our knowledge, this tumor has not been previously described in the nasal septum as a primary lesion. We describe a 39 year-old woman who presented with a 1-year history of nasal obstruction. Results of her physical examination and imaging studies demonstrated a nasoseptal mass. We also discuss the histologic and radiographic characteristics of the chondroid chordoma and compare it with other tumors that arise in the nasoseptal area to highlight the different prognoses and approaches to management. PMID- 12578459 TI - Prophylactic treatment reduces the severity of xerostomia following radiation therapy for oral cavity cancer. PMID- 12578460 TI - Xerostomia prevention after head and neck cancer treatment. PMID- 12578461 TI - Clinical commentary on prophylactic treatment of radiation-induced xerostomia. PMID- 12578462 TI - Pathology quiz case 1. Chronic granulomatous disease (CGD). PMID- 12578463 TI - Pathology quiz case 2. Pemphigus vulgaris (PV). PMID- 12578464 TI - Radiology quiz case 1. Epidural empyema after acute mastoiditis. PMID- 12578465 TI - Radiology quiz case 2. Verrucous carcinoma of the maxillary antrum. PMID- 12578466 TI - EPA ponders pesticide toxicity testing: considers ending moratorium on human data. PMID- 12578467 TI - Researchers say changes needed in recruitment policies for NIH trials. PMID- 12578468 TI - FDA orders estrogen safety warnings: agency offers guidance for HRT use. PMID- 12578472 TI - JAMA patient page. Baseball safety for children. PMID- 12578473 TI - Ginkgo and memory. PMID- 12578474 TI - Ginkgo and memory. PMID- 12578475 TI - Ginkgo and memory. PMID- 12578476 TI - Ginkgo and memory. PMID- 12578477 TI - Ginkgo and memory. PMID- 12578478 TI - Racial differences in rates of traumatic lumbar puncture. PMID- 12578480 TI - Nursing burnout and patient safety. PMID- 12578481 TI - Nursing burnout and patient safety. PMID- 12578482 TI - Nursing burnout and patient safety. PMID- 12578484 TI - Certificate of need regulations and hospital mortality. PMID- 12578486 TI - Ethanol in formularies of US teaching hospitals. PMID- 12578487 TI - Acetylcysteine for prevention of acute deterioration of renal function following elective coronary angiography and intervention: a randomized controlled trial. AB - CONTEXT: The antioxidant acetylcysteine prevents acute contrast nephrotoxicity in patients with impaired renal function who undergo computed tomography scanning. However, its role in coronary angiography is unclear. OBJECTIVE: To determine whether oral acetylcysteine prevents acute deterioration in renal function in patients with moderate renal insufficiency who undergo elective coronary angiography. DESIGN AND SETTING: Prospective, randomized, double-blind, placebo controlled trial conducted from May 2000 to December 2001 at the Grantham Hospital at the University of Hong Kong. PARTICIPANTS: Two hundred Chinese patients aged mean (SD) 68 (6.5) years with stable moderate renal insufficiency (creatinine clearance <60 mL/min [1.00 mL/s]) who were undergoing elective coronary angiography with or without intervention. INTERVENTION: Participants were randomly assigned to receive oral acetylcysteine(600 mg twice per day; n = 102) or matching placebo tablets (n = 98) on the day before and the day of angiography. All patients received low-osmolality contrast agent. MAIN OUTCOME MEASURES: Occurrence of more than a 25% increase in serum creatinine level within 48 hours after contrast administration; change in creatinine clearance and serum creatinine level. RESULTS: Twelve control patients (12%) and 4 acetylcysteine patients (4%) developed a more than 25% increase in serum creatinine level within 48 hours after contrast administration (relative risk, 0.32; 95% confidence interval [CI], 0.10-0.96; P =.03). Serum creatinine was lower in the acetylcysteine group (1.22 mg/dL [107.8 micromol/L]; 95% CI, 1.11-1.33 mg/dL vs 1.38 mg/dL [122.9 micromol/L]; 95% CI, 1.27-1.49 mg/dL; P =.006) during the first 48 hours after angiography. Acetylcysteine treatment significantly increased creatinine clearance from 44.8 mL/min (0.75 mL/s) (95% CI, 42.7-47.6 mL/min) to 58.9 mL/min (0.98 mL/s) (95% CI, 55.6-62.3 mL/min) 2 days after the contrast administration (P<.001). The increase was not significant in the control group (from 42.1 to 44.1 mL/min [0.70 to 0.74 mL/s]; P =.15). The benefit of acetylcysteine was consistent among various patient subgroups and persistent for at least 7 days. There were no major treatment-related adverse events. CONCLUSION: Acetylcysteine protects patients with moderate chronic renal insufficiency from contrast-induced deterioration in renal function after coronary angiographic procedures, with minimal adverse effects and at a low cost. PMID- 12578488 TI - Health and function of patients with untreated idiopathic scoliosis: a 50-year natural history study. AB - CONTEXT: Previous long-term studies of idiopathic scoliosis have included patients with other etiologies, leading to the erroneous conclusion that all types of idiopathic scoliosis inevitably end in disability. Late-onset idiopathic scoliosis (LIS) is a distinct entity with a unique natural history. OBJECTIVE: To present the outcomes related to health and function in untreated patients with LIS. DESIGN, SETTING, AND PATIENTS: Prospective natural history study performed at a midwestern university with outpatient evaluation of patients who presented between 1932 and 1948. At 50-year follow-up, which began in 1992, 117 untreated patients were compared with 62 age- and sex-matched volunteers. The patients' mean age was 66 years (range, 54-80 years). MAIN OUTCOME MEASURES: Mortality, back pain, pulmonary symptoms, general function, depression, and body image. RESULTS: The estimated probability of survival was approximately 0.55 (95% confidence interval [CI], 0.47-0.63) compared with 0.57 expected for the general population. There was no significant difference in the demographic characteristics of the 2 groups. Twenty-two (22%) of 98 patients complained of shortness of breath during everyday activities compared with 8 (15%) of 53 controls. An increased risk of shortness of breath was also associated with the combination of a Cobb angle greater than 80 degrees and a thoracic apex (adjusted odds ratio, 9.75; 95% CI, 1.15-82.98). Sixty-six (61%) of 109 patients reported chronic back pain compared with 22 (35%) of 62 controls (P =.003). However, of those with pain, 48 (68%) of 71 patients and 12 (71%) of 17 controls reported only little or moderate back pain. CONCLUSIONS: Untreated adults with LIS are productive and functional at a high level at 50-year follow-up. Untreated LIS causes little physical impairment other than back pain and cosmetic concerns. PMID- 12578489 TI - Evaluation of safety balls and faceguards for prevention of injuries in youth baseball. AB - CONTEXT: Safety balls and faceguards are widely used in youth baseball, but their effectiveness in reducing injury is unknown. OBJECTIVE: To evaluate the association of the use of faceguards and safety balls and injuries in youth baseball. DESIGN, SETTING, AND PARTICIPANTS: Ecological study using a national database of compensated insurance claims maintained by Little League Baseball Incorporated, combined with data on the number of participants in Little League and data from a census of protective equipment usage for youth aged 5 to 18 years participating in Little League Baseball in the United States during 1997-1999. MAIN OUTCOME MEASURES: Rate of injury and injury rate ratio comparing users with nonusers of protective equipment. RESULTS: A total of 6 744 240 player-seasons of follow-up and 4233 compensated injury claims were available for analysis. The absolute incidence of compensated injury per 100 000 player-seasons was 28.02 (95% confidence interval [CI], 26.76-29.29) for ball-related injury and 2.71 (95% CI, 2.32-3.11) for facial injury. Overall, use of safety balls was associated with a reduced risk of ball-related injury (adjusted rate ratio, 0.77; 95% CI, 0.64-0.93). This reduction was essentially due to 1 type of safety ball, known as the reduced-impact ball (adjusted rate ratio, 0.72; 95% CI, 0.57-0.91). Use of faceguards reduced the risk of facial injury (adjusted rate ratio, 0.65; 95% CI, 0.43-0.98). Metal and plastic guards appeared to be equally effective. Safety balls appeared to be more effective in the minor division (ages 7-12 years) than in the regular division (ages 9-12 years). CONCLUSIONS: Reduced-impact balls and faceguards were associated with a reduced risk of injury in youth baseball. These findings support increased usage of these items; however, it should be noted that the absolute incidence of injury in youth baseball is low and that these equipment items do not prevent all injuries. PMID- 12578490 TI - Variation in public and private supply of pneumococcal conjugate vaccine during a shortage. AB - CONTEXT: In late August 2001, a serious shortage of the heptavalent pneumococcal conjugate vaccine (PCV7) developed in 34 state immunization programs. In September 2001, the Centers for Disease Control and Prevention published revised recommendations advising physicians to prioritize PCV7 to specific groups of children. The effect of the shortage at the practice level is unknown. OBJECTIVE: To determine the variation between public and private markets in the supply of PCV7 and the nature and extent of the PCV7 shortage at the practice level. DESIGN, SETTING, AND PARTICIPANTS: Semistructured interviews with office staff responsible for ordering vaccines at private practices in 12 states were conducted between October 19 and November 2, 2001. MAIN OUTCOME MEASURES: Variation in supply of PCV7 obtained from public sources and through purchase on the private market. Supply was characterized into 3 categories: "no problem," "problem obtaining a consistent supply," and "out of stock." RESULTS: Interviews were completed at 405 practices, representing a response rate of 74%. Overall, 51% of practices reported at least 1 episode of being out of stock of public PCV7 and 64% of private PCV7, with significant state-to-state variation. Only 2 of 12 study states had a substantially higher proportion of practices experiencing out of-stock episodes for public compared with private PCV7, while in 6 states public PCV7 was less frequently out of stock than private PCV7. Only 23% of practices in this study altered their administration policy for private PCV7, while 27% altered their policy for public PCV7. CONCLUSIONS: The distribution and supply of PCV7 varied between public and private supplies and between states during the shortage. Few practices changed their administration schedules in response to revised recommendations. PMID- 12578491 TI - Alcohol consumption and risk of stroke: a meta-analysis. AB - CONTEXT: Observational studies suggest that heavy alcohol consumption may increase the risk of stroke while moderate consumption may decrease the risk. OBJECTIVE: To examine the association between alcohol consumption and relative risk of stroke. DATA SOURCES: Studies published in English-language journals were retrieved by searching MEDLINE (1966-April 2002) using Medical Subject Headings alcohol drinking, ethanol, cerebrovascular accident, cerebrovascular disorders, and intracranial embolism and thrombosis and the key word stroke; Dissertation Abstracts Online using the keywords stroke and alcohol; and bibliographies of retrieved articles. STUDY SELECTION: From 122 relevant retrieved reports, 35 observational studies (cohort or case control) in which total stroke, ischemic stroke, or hemorrhagic (intracerebral or total) stroke was an end point; the relative risk or relative odds and their variance (or data to calculate them) of stroke associated with alcohol consumption were reported; alcohol consumption was quantified; and abstainers served as the reference group. DATA EXTRACTION: Information on study design, participant characteristics, level of alcohol consumption, stroke outcome, control for potential confounding factors, and risk estimates was abstracted independently by 3 investigators using a standardized protocol. DATA SYNTHESIS: A random-effects model and meta-regression analysis were used to pool data from individual studies. Compared with abstainers, consumption of more than 60 g of alcohol per day was associated with an increased relative risk of total stroke, 1.64 (95% confidence interval [CI], 1.39-1.93); ischemic stroke, 1.69 (95% CI, 1.34-2.15); and hemorrhagic stroke, 2.18 (95% CI, 1.48-3.20), while consumption of less than 12 g/d was associated with a reduced relative risk of total stroke, 0.83 (95%, CI, 0.75-0.91) and ischemic stroke, 0.80 (95% CI, 0.67-0.96), and consumption of 12 to 24 g/d was associated with a reduced relative risk of ischemic stroke, 0.72 (95%, CI, 0.57-0.91). The meta regression analysis revealed a significant nonlinear relationship between alcohol consumption and total and ischemic stroke and a linear relationship between alcohol consumption and hemorrhagic stroke. CONCLUSIONS: These results indicate that heavy alcohol consumption increases the relative risk of stroke while light or moderate alcohol consumption may be protective against total and ischemic stroke. PMID- 12578492 TI - Interventions for violence against women: scientific review. AB - CONTEXT: Intimate partner violence is prevalent and is associated with significant impairment, yet it remains unclear which interventions, if any, reduce rates of abuse and reabuse. OBJECTIVE: To systematically review, from the perspective of primary health care, the available evidence on interventions aimed at preventing abuse or reabuse of women. DATA SOURCES: MEDLINE, PsycINFO, CINAHL, HealthStar, and Sociological Abstracts were searched from the database start dates to March 2001 using database-specific key words such as domestic violence, spouse abuse, partner abuse, shelters, and battered women. References of key articles were hand searched. The search was updated in December 2002. STUDY SELECTION: Both authors reviewed all titles and abstracts using established inclusion/exclusion criteria. Twenty-two articles met the inclusion criteria for critical appraisal. DATA EXTRACTION: Following the evidence-based methods of the Canadian Task Force on Preventive Health Care, both authors independently reviewed the 22 included studies using an established hierarchy of study designs and criteria for rating internal validity. Quality ratings of individual studies- good, fair, or poor--were determined based on a set of operational parameters specific to each design category developed with the US Preventive Services Task Force. DATA SYNTHESIS: Screening instruments exist that can identify women who are experiencing intimate partner violence. No study has examined, in a comparative design, the effectiveness of screening when the end point is improved outcomes for women (as opposed to identification of abuse). No high-quality evidence exists to evaluate the effectiveness of shelter stays to reduce violence. Among women who have spent at least 1 night in a shelter, there is fair evidence that those who received a specific program of advocacy and counseling services reported a decreased rate of reabuse and an improved quality of life. The benefits of several other intervention strategies in treating both women and men are unclear, primarily because of a lack of suitably designed research measuring appropriate outcomes. In most cases, the potential harms of interventions are not assessed within the studies reviewed. CONCLUSIONS: Much has been learned in recent years about the epidemiology of violence against women, yet information about evidence-based approaches in the primary care setting for preventing intimate partner violence is seriously lacking. The evaluation of interventions to improve the health and well-being of abused women remains a key research priority. PMID- 12578493 TI - Interventions for intimate partner violence against women: clinical applications. AB - Sufficient evidence regarding the effectiveness of physician screening, counseling, and referral for intimate partner violence is lacking. However, even in the absence of sufficient evidence, many medical organizations recommend that physicians make efforts to identify and refer patients who have experienced abuse. This article presents 3 cases that illustrate the interaction between patients experiencing violence in intimate relationships and physicians and other members of the health care team. Suggestions for care are based on guidelines and evidence where available. Resources to assist physicians with referral to appropriate services also are provided. PMID- 12578494 TI - Prevention of contrast nephropathy. PMID- 12578495 TI - Sizing up scoliosis. PMID- 12578496 TI - MSJAMA. Once upon a time. PMID- 12578498 TI - MSJAMA. Watching the world go by. PMID- 12578500 TI - MSJAMA. Colostomy. PMID- 12578502 TI - MSJAMA. Pain and forgetting. PMID- 12578503 TI - From the Centers for Disease Control and Prevention. Outbreaks of Salmonella serotype enteritidis infection associated with eating shell eggs--United States, 1999-2001. PMID- 12578504 TI - From the Centers for Disease Control and Prevention. Blood safety monitoring among persons with bleeding disorders--United States, May 1998-June 2002. PMID- 12578505 TI - From the Centers for Disease Control and Prevention. Tobacco use among middle and high school students--New Hampshire, 1995-2001. PMID- 12578506 TI - A systematic review of alternative therapies in the irritable bowel syndrome. AB - The irritable bowel syndrome is a common disorder associated with a significant burden of illness, poor quality of life, high rates of absenteeism, and high health care utilization. Management can be difficult and treatment unrewarding; these facts have led physicians and patients toward alternative therapies. We explored a variety of treatments that exist beyond the scope of commonly used therapies for irritable bowel syndrome. Guarded optimism exists for traditional Chinese medicine and psychological therapies, but further well-designed trials are needed. Oral cromolyn sodium may be useful in chronic unexplained diarrhea and appears as effective as and safer than elimination diets. The roles of lactose and fructose intolerance remain poorly understood. Alterations of enteric flora may play a role in irritable bowel syndrome, but supporting evidence for bacterial overgrowth or probiotic therapy is lacking. PMID- 12578507 TI - Respiratory consequences of rhinovirus infection. AB - Rhinoviruses, a genus of the family Picornaviridae, are the cause of more than 50% of respiratory tract infections. Complications of rhinovirus infections, which include otitis media, sinusitis, exacerbations of asthma, and other pulmonary diseases, can be significant in certain populations. Reverse transcriptase-polymerase chain reaction has allowed the identification of rhinoviruses and led to a greater appreciation of the role of this pathogen in upper and lower respiratory tract disease. Furthermore, antiviral agents with broad activity against rhinoviruses have recently been developed, have undergone clinical trials, but have not been approved for clinical use. By altering the clinical course of picornavirus infections, it may be possible to minimize their potential adverse consequences. PMID- 12578508 TI - Prevalence of celiac disease in at-risk and not-at-risk groups in the United States: a large multicenter study. AB - BACKGROUND: Celiac disease (CD) is an immune-mediated enteropathic condition triggered in genetically susceptible individuals by the ingestion of gluten. Although common in Europe, CD is thought to be rare in the United States, where there are no large epidemiologic studies of its prevalence. The aim of this study was to determine the prevalence of CD in at-risk and not-at-risk groups in the United States. METHODS: Serum antigliadin antibodies and anti-endomysial antibodies (EMA) were measured. In EMA-positive subjects, human tissue transglutaminase IgA antibodies and CD-associated human leukocyte antigen DQ2/DQ8 haplotypes were determined. Intestinal biopsy was recommended and performed whenever possible for all EMA-positive subjects. A total of 13 145 subjects were screened: 4508 first-degree and 1275 second-degree relatives of patients with biopsy-proven CD, 3236 symptomatic patients (with either gastrointestinal symptoms or a disorder associated with CD), and 4126 not-at-risk individuals. RESULTS: In at-risk groups, the prevalence of CD was 1:22 in first-degree relatives, 1:39 in second-degree relatives, and 1:56 in symptomatic patients. The overall prevalence of CD in not-at-risk groups was 1:133. All the EMA-positive subjects who underwent intestinal biopsy had lesions consistent with CD. CONCLUSIONS: Our results suggest that CD occurs frequently not only in patients with gastrointestinal symptoms, but also in first- and second-degree relatives and patients with numerous common disorders even in the absence of gastrointestinal symptoms. The prevalence of CD in symptomatic patients and not at-risk subjects was similar to that reported in Europe. Celiac disease appears to be a more common but neglected disorder than has generally been recognized in the United States. PMID- 12578509 TI - Trends in treatment and outcomes for acute stroke patients in Ontario, 1992-1998. AB - BACKGROUND: Several interventions have been shown to be of benefit to patients with stroke (hereafter referred to as stroke patients) in clinical trials, but the net effect of these interventions in the general stroke population has not been established. The purpose of this study was to evaluate temporal trends in the characteristics, treatments, and outcomes of acute stroke patients in the province of Ontario. METHODS: We conducted a population-based retrospective cohort study using linked administrative databases of all 91 419 patients discharged with a most-responsible diagnosis of acute stroke from acute care hospitals in Ontario from April 1, 1992, to March 31, 1999 (fiscal years 1992 1998). RESULTS: The average age and proportion of stroke patients with co existing diseases increased over time. The proportion of elderly patients 65 years and older who received warfarin sodium (Coumadin) and statins increased during the study period (14.6% to 19.6% [P =.001] and 2.7% to 15.0% [P<.001], respectively). Declines in the median length of stay (11 to 8 days [P<.001]) and risk-adjusted in-hospital mortality (21.9% to 18.9% [P<.001]) were significant, but the 30-day mortality rates for acute stroke stayed relatively constant (19.7% to 19.0% [P =.18]). We found a moderate decline in risk-adjusted 1-year mortality (34.1% to 32.0% [P<.001]) and stroke readmission rates (12.1% to 9.9% [P =.001]). CONCLUSIONS: Improvements in the outcomes of stroke patients have occurred in Ontario during the 1990s, despite an increasing proportion of elderly stroke patients with multiple comorbidities. Increasing use of secondary prevention medications may explain this trend. PMID- 12578510 TI - Depression and adverse drug reactions among hospitalized older adults. AB - BACKGROUND: Depression is a common disorder among hospitalized older adults, and it has been associated with adverse outcomes during hospital stays, including increased risk of morbidity and mortality and reduced recovery rates from illness and disability. The aim of this study was to assess whether depression may be a risk factor for adverse drug reactions (ADRs) among hospitalized older adults. METHODS: A total of 3134 older patients admitted to 23 hospitals throughout Italy between May 1 and June 30 and September 1 and October 31, 1998, entered the study. Patients with a short-form Geriatric Depression Scale score of 5 or greater were considered depressed. Adverse drug reactions observed during hospital stays and classified as definite or probable on the basis of the Naranjo algorithm were considered for this study. RESULTS: During the hospital stays, 192 ADRs were identified in 183 patients (5.8% of the sample). Cardiovascular and arrhythmic complications (20.3% of all ADRs) were the most frequent ADRs, followed by gastrointestinal (18.8%), dermatologic and allergic (12.5%), hemorrhagic (11.5%), and electrolyte (9.9%) disturbances. Adverse drug reactions were recorded in 101 (7.4%) of 1363 depressed patients and in 82 (4.6%) of 1771 nondepressed patients (P =.001). After adjusting for potential confounders, depression was associated with a significantly higher rate of ADRs (odds ratio, 1.58; 95% confidence interval, 1.14-2.20; P =.006). This effect seemed more pronounced in women (odds ratio, 1.85; 95% confidence interval, 1.16-2.95) than in men (odds ratio, 1.38; 95% confidence interval, 0.85-2.34). CONCLUSION: In older hospitalized patients, depression seems to be associated with a greater occurrence of ADRs. PMID- 12578511 TI - Major dietary patterns and the risk of colorectal cancer in women. AB - BACKGROUND: Several foods and nutrients have been implicated in the development of colon and rectal cancers. In this study, we prospectively assessed the associations between major dietary patterns and the risks of these 2 cancers in women. METHODS: Using dietary information collected in 1984, 1986, 1990, and 1994 from 76 402 women aged 38 to 63 years without a history of cancer in 1984, we conducted factor analysis and identified 2 major dietary patterns: "prudent" and "Western." We calculated factor scores for each participant and examined prospectively the associations between dietary patterns and colon and rectal cancer risks. RESULTS: The prudent pattern was characterized by higher intakes of fruits, vegetables, legumes, fish, poultry, and whole grains, while the Western pattern, by higher intakes of red and processed meats, sweets and desserts, french fries, and refined grains. During 12 years of follow-up, we identified 445 cases of colon cancer and 101 cases of rectal cancer. After adjusting for potential confounders, we observed a relative risk for colon cancer of 1.46 (95% confidence interval, 0.97-2.19) when comparing the highest with the lowest quintiles of the Western pattern (P value for trend across quintiles, .02). The prudent pattern had a nonsignificant inverse association with colon cancer (relative risk for fifth quintile compared with the first, 0.71; 95% confidence interval, 0.50-1.00; P for trend across quintiles, .31). We did not observe any significant association between dietary patterns and rectal cancer. CONCLUSION: We found a significant positive association between the Western dietary pattern and the risk of colon cancer. PMID- 12578512 TI - Pneumonia: still the old man's friend? AB - BACKGROUND: Hospital mortality of patients admitted with community-acquired pneumonia (CAP) has been well described. However, the long-term survival of those discharged alive is less clear. We sought to determine long-term survival of patients hospitalized with CAP and compare the outcome with controls hospitalized for reasons other than CAP. METHODS: We performed a matched case-control analysis using the Medicare hospital discharge database from the first quarter of 1997. We compared all Medicare recipients 65 years or older hospitalized with CAP and controls matched for age, sex, and race hospitalized for reasons other than CAP. We measured 1-year mortality determined from the Medicare Beneficiary Entitlement file and the Social Security Administration. RESULTS: We identified 158 960 CAP patients and 794 333 hospitalized controls. Hospital mortality rates for the CAP cohort and hospitalized controls were 11.0% and 5.5%, respectively (P<.001). One year mortality rates for the CAP cohort and hospitalized controls were 40.9% and 29.1%, respectively (P<.001). One-year mortality rates in hospital survivors of the CAP and control cohorts were 33.6% and 24.9%, respectively (P<.001). The difference in mortality between the CAP and control cohorts was not explained by underlying disease. Standardized against the general population, the risk of death for both cohorts decreased monthly but was still elevated 1 year after hospital discharge. The standardized mortality ratio was 2.69 (95% confidence interval, 2.47-2.93) for CAP patients and 1.93 (95% confidence interval, 1.79 2.08) for hospital controls. CONCLUSIONS: Almost half of all elderly patients admitted for CAP die in the subsequent year, with most deaths occurring after hospital discharge. The mortality is considerably higher than that of either the general population or a control population hospitalized for reasons other than CAP. PMID- 12578513 TI - Improving the quality of care for patients with pneumonia in very small hospitals. AB - BACKGROUND: Despite the publication of guidelines for the management of pneumonia, significant variation in care continues to exist. While there have been several published reports of quality improvement projects for pneumonia, there are few data on the effectiveness of these efforts in small hospitals. The purpose of this study was to demonstrate that a project implemented by a quality improvement organization in small hospitals would lead to an improvement in care that could not be accounted for by secular trends in the management of pneumonia. METHODS: Medicare-insured hospital admissions for pneumonia were reviewed from 20 small hospitals in Oklahoma (intervention group) at baseline and after feedback. Project intervention included onsite feedback presentations to the medical staff, samples of performance improvement materials, and comparative measures of performance of predefined quality indicators. A second group of 16 demographically similar hospitals (control group) was selected for review during the same 2 periods. These hospitals subsequently underwent an identical intervention with a follow-up assessment. RESULTS: Statistically significant improvements in process measures were demonstrated in the intervention hospitals, including performance of a sputum (P<.01) and blood (P<.001) cultures, antibiotic administration within 4 hours of hospital admission (P<.001), and administration of the first dose of antibiotic in the emergency department (P<.001). These measures in the control hospitals did not change significantly (P =.93, .08, .79, and .52, respectively) during the 2 periods. CONCLUSIONS: Improvements in processes of care achieved by the intervention hospitals resulted from activities initiated because of participation in a quality improvement organization-directed project. This study demonstrated the effectiveness of quality improvement activities in very small hospitals. PMID- 12578514 TI - The importance of indirect costs in primary cardiovascular disease prevention: can we save lives and money with statins? AB - BACKGROUND: The losses in productivity due to cardiovascular disease (CVD) are substantial but rarely considered in health economic analyses. We compared the cost-effectiveness of lipid level modification in the primary prevention of CVD with and without these indirect costs. METHODS: We used the Cardiovascular Life Expectancy Model to estimate the long-term benefits and cost-effectiveness of lipid level modification with atorvastatin calcium, including 28% and 38% reductions in total cholesterol and low-density lipoprotein cholesterol levels, respectively, and a 5.5% increase in high-density lipoprotein cholesterol level. The direct costs included all medical care costs associated with CVD. The indirect costs represented the loss of employment income and the decreased value of housekeeping services after different manifestations of CVD. All costs were expressed in 2000 Canadian dollars. RESULTS: When only direct medical care costs were considered, the incremental cost-effectiveness ratios for lifelong therapy with atorvastatin calcium, 10 mg/d, were generally positive, ranging from a few thousand to nearly $20 000 per year of life saved. When the societal point of view was adopted and indirect costs were included, the total costs were generally negative, representing substantial cost savings (up to $50 000) and increased life expectancy for most groups of individuals. CONCLUSIONS: Lipid therapy with statins can reduce CVD morbidity and mortality as demonstrated in a number of clinical trials. Adding the indirect CVD costs associated with productivity losses at work and home can result in forecasted cost savings to society as a whole such that lipid therapy could potentially save lives and money. PMID- 12578515 TI - Sedative use in the last week of life and the implications for end-of-life decision making. AB - BACKGROUND: The use of sedation at the end of life has aroused ethical controversy, attracting accusations of hastening death by gradually increasing sedative doses. The doctrine of double effect has been introduced as an ethical defense. This study aimed to determine how sedative doses change at the end of life and how often the doctrine of double effect might be relevant. METHODS: Case note review was performed of 237 consecutive patients who died in a specialist palliative care unit. Sedative dose changes during the last week of life were noted and survival from admission was compared between groups of patients receiving no sedation, sedation for 7 days, or a commencement of sedation in the last 48 hours of life. There was detailed review of notes from patients who received a marked increase in sedative dose to explore the applicability of the doctrine of double effect. RESULTS: Sedation was given to 48% of patients. Of these, 13% received sedatives for 7 days or more, while 56% commenced sedative use only in the last 48 hours of life. The groups receiving no sedation or sedation for less than 48 hours had the shortest survival from admission (mean, 14.3 and 14.2 days), whereas the 7-day sedation group survived for a mean of 36.6 days (P<.001). Sedative use and dose increased toward the end of life, but the detailed case note review disclosed only 2 cases where the doctrine of double effect may have been implicated. CONCLUSION: Sedative dose increases in the last hours of life were not associated with shortened survival overall, suggesting that the doctrine of double effect rarely has to be invoked to excuse sedative prescribing in end-stage care. PMID- 12578516 TI - Outcomes of chronic heart failure. AB - BACKGROUND: Outcomes related to chronic heart failure (HF) remain relatively poor, despite advances in pharmacological therapy and medical and nursing care. Experts agree that outpatient care may be among the factors that affect HF outcomes. We hypothesized that the method by which outpatient care is delivered may affect outcomes in this patient population. METHODS: A prospective, randomized design was used to compare HF outcomes from 216 patients randomized to 1 of 2 home health care delivery methods for 3 months after discharge. Care was delivered by the home nurse visit (HNV) or the nurse telemanagement (NTM) method. In the latter, patients used transtelephonic home monitoring devices to measure their weight, blood pressure, heart rate, and oxygen saturation. These data were transmitted daily to a secure Internet site. An advanced-practice nurse worked collaboratively with a cardiologist and subsequently treated patients via the telephone. Both delivery methods used the same HF-specific clinical guidelines to direct care. Outcomes include HF readmissions and length of stay, anxiety, depression, self-efficacy, and quality of life. Data were primarily tested using a 2-group analysis of variance (ANOVA). We used a repeated-measures ANOVA to conduct preintervention-postintervention analyses. RESULTS: After 3 months, patients in the NTM group (n = 108; mean +/- SD age, 62.9 +/- 13.2 years; 83% African American; 64% female) had fewer HF readmissions (13 vs 24; P150 micro mol/L) had a sensitivity of 12.6% and a specificity of 99.9% for the detection of renal failure. For the detection of severe renal failure, the sensitivity was 45.5%, with a 99.1% specificity. Only 15 (27.3%) of the 55 patients with severe renal failure were referred to a nephrologist. Moreover, 34 (85%) of the 40 nonreferred patients with severe renal failure were incompletely evaluated regarding the metabolic complications associated with kidney dysfunction. CONCLUSION: Serum creatinine is a poor screening test for renal failure in elderly patients, leading to marked underinvestigation and underrecognition of renal failure in this population. PMID- 12578518 TI - Validity of the JNC VI recommendations for the management of hypertension in a general population of Japanese elderly: the Hisayama study. AB - BACKGROUND: It is not known whether the treatment recommendations presented in the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure are applicable to the Japanese elderly population. METHODS: We followed up 588 cardiovascular disease-free residents of a Japanese community who were 60 years or older from November 1, 1961, through October 31, 1993. Treated hypertensive patients were excluded from the analysis. During this period, CVD occurred in 179 subjects. The incidences were estimated by the pooling of repeated observations method. RESULTS: The age- and sex-adjusted incidences of cardiovascular disease significantly increased with elevated blood pressure levels. The hazard ratio for stage 3 hypertension was 5.34 (95% confidence interval, 2.66-10.71; P<.001) compared with optimal blood pressure after adjustment for other covariates. Among subjects aged 60 to 79 years, the incidences for stages 1 through 3 hypertension were significantly higher than for those with optimal and normal blood pressure. In comparison, among those 80 years or older, the incidence was significantly higher only in patients with stage 3 hypertension. We further estimated the incidences according to the risk stratification system. In the younger elderly subjects, the incidences increased with rising blood pressure levels in each risk stratum. Similar relationships were not observed among the older elderly subjects. CONCLUSIONS: Our findings demonstrate that the recommendations of the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure were potentially applicable to the Japanese elderly subjects 79 years or younger. Based on our findings, however, hypertension might not be a risk factor for cardiovascular disease among very old hypertensive patients with advanced atherosclerosis. PMID- 12578519 TI - Intentional clonidine patch ingestion by 3 adults in a detoxification unit. AB - Clonidine hydrochloride is an alpha(2)-adrenergic agonist commonly prescribed as an antihypertensive agent. It is also useful in ameliorating symptoms associated with opiate withdrawal and is frequently used in the detoxification setting. We report an episode in which 3 normotensive patients each ingested a single 0.1-mg clonidine patch in a detoxification unit. PMID- 12578520 TI - Reporting the NAFT major bleeding rates in context: reviewers should include site investigator-classified rates. PMID- 12578522 TI - The use of niacin in diabetes mellitus. PMID- 12578524 TI - The impact of coffee drinking. PMID- 12578526 TI - Continued need for placebo in many cases, even when there is effective therapy. PMID- 12578528 TI - Oestrogen receptor knockout mice: roles for oestrogen receptors alpha and beta in reproductive tissues. AB - Oestrogen is an essential component of female reproduction, with well characterized functions in the uterus, ovaries, mammary gland and hypothalamic pituitary axis. The mechanism of oestrogen action involves mediation of the rate of transcription by nuclear-localized oestrogen receptor molecules. Two oestrogen receptors are present in mouse tissues, oestrogen receptors alpha and beta. Each receptor exhibits differential tissue expression patterns. Mouse models with genetically engineered disruption or 'knockout' of the oestrogen receptors have been developed. Characterization of the resulting defects in reproductive tissues as well as alterations in physiological and genomic responses has given insight into the receptor-mediated effects of oestrogen in reproduction. Oestrogen receptor alpha knockout females are infertile because they are anovulatory, have disruption in LH regulation and have uteri that are insensitive to oestrogen. In contrast, oestrogen receptor beta knockout females are sub-fertile and primarily lack efficient ovulatory function. Mice with deletion of both oestrogen receptors alpha and beta are similar to those lacking oestrogen receptor alpha only, but exhibit a unique ovarian pathology. These observed phenotypes elucidate the relative roles of the oestrogen receptors in reproductive functions of female rodents. PMID- 12578529 TI - A conceptual model of the influence of stress on female reproduction. AB - Intriguingly, similar neurotransmitters and nuclei within the hypothalamus control stress and reproduction. GnRH neurone recruitment and activity is regulated by a balance between stimulation, suppression and permissiveness controlled by noradrenaline, neuropeptide Y and serotonin from the brain stem, impact from glutamate in the medial preoptic area and neuropeptide Y in the arcuate nucleus, in opposition to the restraining influences of gamma aminobenzoic acid within the medial preoptic area and opioids from the arcuate nucleus. Stress also activates neuropeptide Y perikarya in the arcuate nucleus and brain stem noradrenaline neurones. The latter project either indirectly, via the medial preoptic area, or directly to the paraventricular nucleus to release corticotrophin releasing hormone (CRH) and arginine vasopressin (AVP). Within the medial preoptic area, GnRH neurones synapse with CRH and AVP axons. Stimulation of CRH neurones in the paraventricular nucleus also activates gamma-aminobenzoic acid and opioid neurones in the medial preoptic area and reduces GnRH cell recruitment, thereby decreasing GnRH pulse frequency. Oestradiol enhances stress induced noradrenaline suppression of LH pulse frequency but when applied in the paraventricular nucleus or brain stem, and not in the medial preoptic area or arcuate nucleus. The importance of CRH and AVP in the medial preoptic area needs confirming in a species other than the rat, which uses adrenal activation to time the onset of the GnRH surge. Another stress-activated pathway involves the amygdala and bed of the nucleus stria terminalis, which contain CRH neurones and accumulate gamma-aminobenzoic acid during stress. PMID- 12578530 TI - Age-related uterine and ovarian hypertrophy in FSH receptor knockout and FSHbeta subunit knockout mice. AB - Female mice in which the gene encoding the follicle-stimulating hormone FSH receptor (FSHR) knockout (KO) or its ligand (FSHbetaKO) have been disrupted were infertile. Ovaries of these mice were significantly smaller than those of heterozygous littermates but significantly larger than those of hypogonadal mice of the same age. Uterine masses in all three mutants were <6 mg, significantly reduced compared with heterozygous mice. At 1 year of age uterine mass had increased to >12 mg in 63% of FSHRKO females and 88% of FSHbetaKO females. Despite the increase in uterine size there was no evidence of contractility: uteri were flaccid and unresponsive to electrical or pharmacological stimulation. In most females in which uterine growth had occurred there was evidence of ovarian growth with hypertrophy of the interstitial tissue, occurrence of ovarian cysts and epithelial and tubular inclusions. There was no evidence of uterine or ovarian hypertrophy in hypogonadal (hpg) mice at any age or in 1 year old females in which the FSH mutations were bred onto the hpg background. There was an inverse correlation of plasma LH concentrations and uterine mass in 1 year old mutant females with uterine hypertrophy. Ovariectomy of both FSHRKO and FSHbetaKO females with large uteri resulted in decreased uterine mass and increased plasma concentration of LH. The number of mice with ovarian pathology, reminiscent of the serous ovarian adenocarcinomas found in humans, was significantly greater in the FSHbetaKO mice, indicating that the presence of an intact FSH receptor on ovarian cells of FSHbetaKO females may allow constitutive basal stimulation of the ovary, which is absent in mice lacking FSH receptors. PMID- 12578531 TI - Motility activation and second messenger signalling in spermatozoa from rat cauda epididymidis. AB - This study examined molecular mechanisms involved in the activation of motility in spermatozoa from the cauda epididymidis of rats. A 1.05-fold dilution of semen from the cauda epididymidis with 300 mmol sucrose l(-1) did not activate motility in spermatozoa. Addition of dibutyryl cAMP, pentoxifylline or Ca(2+) to the sucrose activated motility in the short term (<30-60 min). A fivefold dilution of semen from the cauda epididymidis with a modified Tyrode's medium (BWW) activated and sustained vigorous motility that could not be attenuated with kinase inhibitors. This motility was associated with a transient increase in intracellular cAMP during the first 60 s of activation. Lower motility was activated in Ca(2+)-deficient media but this was not associated with an increase in cAMP. A fivefold dilution with plasma from the cauda epididymidis did not activate motility. The addition of Ca(2+) to the sucrose induced an increase in cAMP of similar duration but lower magnitude to that associated with dilution in BWW. The results from this study indicate that the cAMP and Ca(2+) signal transduction pathways are involved in activation of sperm motility, and that the increase in intracellular cAMP in rat spermatozoa from the cauda epididymidis undergoing motility activation is Ca(2+)-dependent. This is the first study to report a Ca(2+)-dependent increase in cAMP associated with motility activation in immotile mammalian spermatozoa. In light of these data, a model is proposed whereby cAMP and Ca(2+) act as synarchic messengers, initiating a signal transduction cascade, which is independent of protein kinase A-mediated phosphorylation of flagella proteins in immotile spermatozoa from the cauda epididymidis. PMID- 12578532 TI - Postpartum uterine involution in sheep: histoarchitecture and changes in endometrial gene expression. AB - After parturition, the uterus undergoes marked remodelling during involution; however, little is known of the hormonal, cellular and molecular mechanisms that regulate this process. The working hypothesis used in this study is that return of the ovine uterus to a non-pregnant state involves termination of a hormonal servomechanism that regulates endometrial gland morphogenesis and function during pregnancy. Suffolk ewes were ovariohysterectomized on postpartum days 1, 7, 14 or 28. Serum concentrations of oestradiol were high at parturition, declined to postpartum day 4, peaked on postpartum day 6, and then declined and remained low thereafter. Progesterone was undetectable in plasma from ewes post partum. Uterine wet mass and horn length decreased after postpartum day 1, but ovarian mass did not change. Residual placental cotyledons were present in the maternal caruncles on postpartum days 1 and 7 and were extruded by postpartum day 14 as plaques that were resorbed by postpartum day 28. The width of the total endometrium, stratum compactum, stratum spongiosum and myometrium, as well as endometrial gland density, decreased after parturition. Most apoptotic cells in the involuting uterus were large, vacuolated and located between the endometrial glandular epithelial cells on postpartum days 1 and 7. Immunofluorescence analyses identified both T and B cells within the glandular epithelium on postpartum day 1. Cell proliferation was detected in the luminal epithelium and glandular epithelium on postpartum days 1 and 7. On postpartum day 1, expression of oestrogen receptor alpha (ERalpha) was not detected in luminal epithelium and was low in glandular epithelium, but ERalpha was present in epithelia thereafter. Progesterone receptor (PR) protein was not detected in endometrial epithelia on postpartum day 1, but was detected in the glandular epithelium thereafter. Between postpartum days 1 and 7, ERalpha and PR protein increased substantially in the endometrial glandular epithelium. On postpartum days 1-28, abundant expression of oxytocin receptor mRNA was detected in endometrial luminal epithelium and superficial to the middle glandular epithelium. Prolactin receptor (PRLR) mRNA was detected in glandular epithelium on all postpartum days, whereas mRNA for uterine milk protein (UTMP), an index of secretory capacity of glandular epithelium, was present only on postpartum day 1. Collectively, these results indicate that uterine involution in ewes involves remodelling of both caruncular and intercaruncular areas of the uterine wall and termination of differentiated uterine gland functions characteristic of pregnancy. PMID- 12578533 TI - Inhibitory effects of formoterol on lipopolysaccharide-induced premature delivery through modulation of proinflammatory cytokine production in mice. AB - Previous studies have demonstrated that formoterol, a beta2-adrenoceptor agonist, has potent tocolytic effects in rats. The aim of the present study was to determine whether formoterol treatment affects proinflammatory cytokine production in a lipopolysaccharide (LPS)-treated premature delivery mouse model. Formoterol was continuously administered by osmotic pump and the number of fetuses in the uteri were counted. Samples of amniotic fluid and plasma were collected 8 and 16 h after systemic administration of LPS. LPS induced premature delivery and an increase in prostaglandin F(2alpha) (PGF(2alpha)), interleukin 1alpha (IL-1alpha), IL-6 and IL-10 in the amniotic fluid, and an increase in IL-6 in plasma. Formoterol blocked all changes except the increase in IL-10. These data indicate that formoterol exerts inhibitory effects on proinflammatory cytokine production, and these effects may play an important role in the prevention of premature delivery. PMID- 12578534 TI - Ovarian function in ewes after treatment with mifepristone early during the oestrous cycle. AB - The aim of this study was to determine whether endogenous progesterone regulates synthesis and secretion of luteal oxytocin. In Expt 1, mature ewes (n = 5 per group) were assigned randomly to control or mifepristone (RU486) treatment groups. Ewes were injected s.c. twice a day with vehicle or 10 mg RU486 on days 5 7 of the oestrous cycle (oestrus = day 0). On day 8, after an i.v. injection with prostaglandin F(2alpha) (250 microg cloprostenol), venous blood samples were collected at frequent intervals to determine plasma oxytocin concentrations. Plasma oxytocin concentrations of RU486-treated ewes were not significantly different from those of control ewes. In Expt 2, ewes were injected s.c. each day with vehicle or 175 mg RU486 on days 2-5 of the oestrous cycle followed by administration of prostaglandin F(2alpha) on day 6. Four of five RU486-treated ewes showed 'split-oestrus' (oestrous behaviour for 36 h and then again at 84-108 h after the onset of initial oestrus). There was no significant difference in mean plasma oxytocin or progesterone concentrations between treatment groups. The mean masses of mature corpora lutea from control and RU486-treated ewes on day 6 of the oestrous cycle did not differ significantly (394.8 +/- 28.8 versus 319.5 +/- 48.3 mg). RU486-treated ewes contained mature corpora lutea, new corpora lutea (two of four ewes) and preovulatory follicles (>or= 10 mm, two of four ewes). The average interoestrous interval for RU486-treated ewes was 9 days more than that for control animals (26.2 +/- 2.9 versus 17 +/- 0.5 days; P < 0.025). PMID- 12578535 TI - Capillary angiogenesis and degeneration in bovine ovarian antral follicles. AB - Angiogenesis and capillary degeneration are both evident during ovarian follicle growth. However, the characteristics and distribution of thecal capillary proliferative and degenerative structures have not been fully defined. Indeed, the role of thecal microvasculature changes in follicular atresia is still a matter of debate. The present study examined the distribution of thecal capillary changes occurring during follicular growth and related the changes to capillary morphology (by scanning electron microscopy, SEM, on bovine ovarian corrosion casts) with the incidence of capillary apoptosis (TdT-mediated dUTP nick end labelling, TUNEL) and follicular status (as confirmed by follicular fluid steroid concentrations). SEM demonstrated well-perfused vascular plexuses of small to large antral follicles with structural and functional changes to capillaries. Angiogenesis was evident mainly in the apical part of the inner capillary layer of medium follicles and the middle or basal part of the inner capillary layer of dominant follicles that exhibited high oestradiol:progesterone ratios. Degenerative capillaries were observed mainly in the outer vascular layers of small follicles, and in the inner and outer vascular layers of medium antral follicles. Although apoptotic structures were present only in the outer capillaries of the theca interna of morphologically healthy antral follicles, atretic follicles showed apoptotic structures in both the outer and inner thecal capillary layers. These results show that angiogenesis increases during bovine follicular growth and occurs unevenly in different inner theca regions of the follicles. The differential angiogenic and degenerative response of theca interna capillaries may reflect differences in the microenvironment of the follicles, which in turn determine the fate of the follicles (continued growth versus atresia). PMID- 12578536 TI - Effect of chronic treatment with the gonadotrophin-releasing hormone agonist azagly-nafarelin on basal concentrations of LH in prepubertal bulls. AB - Administration of GnRH agonist for an extended period inhibits pulsatile LH release but enhances testicular function of bulls. The mechanism whereby long term administration of GnRH agonist enhances testosterone concentration in the blood of bulls has not been determined. The aim of this study was to determine whether chronic treatment with the GnRH agonist, azagly-nafarelin, increases blood concentrations of LH and FSH in prepubertal bulls. Two different doses of the GnRH agonist were administered via Alzet mini-osmotic pumps for 28 days. Blood samples were collected at 20 min intervals for 24 h at days 2, 13 and 25 of treatment. Agonist-treated groups had reduced testosterone pulse frequency (P < 0.05) and increased mean and basal concentrations of testosterone (P < 0.05) compared with untreated control bulls. Basal LH concentrations were higher in agonist-treated bulls during all three periods (P < 0.05) and overall (1 ng ml( 1) higher, compared with control bulls; P < 0.001). Frequency of LH pulses in the agonist-treated groups was reduced to less than one pulse in 24 h. Agonist treated bulls tended to have (P < 0.10) or had (P < 0.05) a slight but significant increase in blood FSH concentration. In conclusion, the higher blood testosterone concentration in bulls after prolonged treatment with GnRH agonist may result, at least in part, from changes in the testes induced by enhanced basal concentration of LH. PMID- 12578537 TI - Studies on sperm storage in the vas deferens of the spinifex hopping mouse (Notomys alexis). AB - The cauda epididymidis, with its relatively cool temperature (32-35 degrees C), is considered to be the main site of sperm storage in male mammals. However, in the adult male spinifex hopping mouse, Notomys alexis, similar numbers of spermatozoa are found in the vas deferens to those in the cauda epididymidis. The present study shows that, unlike in the laboratory mouse in which spermatozoa of the vas deferens are found mainly in the epididymal region of the duct, spermatozoa in the hopping mouse are localized mainly to the middle and urethral regions of the vas deferens which lies in the inguinal and lower abdominal region of the body cavity. After ligation of the vas deferens close to its connection with the epididymis, many spermatozoa in the vas deferens retain the potential for motility for up to 2 weeks, indicating that the viability of spermatozoa is not compromised by being restricted to core body temperature. This urethral region of the vas deferens, in which spermatozoa reside, has a highly divergent structural organization compared with that of common laboratory rodents in which there is an expanded lumen with a network of epithelial folds. Ultrastructural observations of the cells lining the duct indicate that there are not any marked differences in morphology compared with the cells lining the duct in common laboratory murids, but the infoldings of the vas deferens of the hopping mouse are highly vascular which might facilitate supply of oxygen and nutrients to the spermatozoa residing in the lumen. PMID- 12578539 TI - Selection of highly fertilization-competent bovine spermatozoa through adhesion to the Fallopian tube epithelium in vitro. AB - Mammalian spermatozoa undergo a marked reduction in number during their journey through the female reproductive tract. One of the checkpoints in the selection of fertilizing spermatozoa may be the transient adhesion to the Fallopian tube epithelium, an event previously shown to play a key role in sperm storage. Bovine spermatozoa adhering to the Fallopian tube epithelium in vitro may be synchronously released by sulphated glycoconjugates. In the present study, experiments were designed to quantify the number of spermatozoa selected through adhesion, and to compare the zona pellucida (ZP) binding and fertilization competence of the initial sperm suspension versus the bound and unbound sperm subpopulations. Results showed that: (1) a fraction accounting for about 30% of the initial sperm suspension was selected by in vitro adhesion to oviductal epithelial cell monolayers; (2) selected spermatozoa, collected after heparin induced release, had a significantly superior ZP binding and fertilization competence (mean +/- SD: 110 +/- 28 bound spermatozoa per oocyte; % cleavage, mean +/- SEM: 89 +/- 4) compared with both the initial sperm suspension (45 +/- 10 bound spermatozoa per oocyte, P < 0.001; % cleavage: 69 +/- 3, P < 0.05) and the unselected subpopulation (30 +/- 4 bound spermatozoa per oocyte, P < 0.001; % cleavage: 58 +/- 3, P < 0.01). These findings support the hypothesis that binding to oviductal cells is not only beneficial for sperm survival but also represents a crucial step for the selection of spermatozoa endowed with superior fertilization competence. PMID- 12578538 TI - Steroidogenic responses of pig corpora lutea to insulin-like growth factor I (IGF I) throughout the oestrous cycle. AB - This study was designed to investigate the roles of insulin-like growth factor I (IGF-I), IGF-type I receptor (IGF-IR) and IGF-binding proteins (IGFBPs) in regulating progesterone secretion by pig corpora lutea during the oestrous cycle, and the signal transduction pathways involved in mediating the steroidogenic actions of IGF-I. Corpora lutea were collected on days 4, 7, 10, 13 and 15 or 16 of the oestrous cycle, enzyme dissociated and the luteal cells were cultured for 24 h in Medium 199 with IGF-I (0-100 ng ml(-1)), long R(3)-IGF-I (0-100 ng ml( 1)), anti-IGF-I (Sm 1.2B; 0-10 microg ml(-1)), anti-IGF-IR (alphaIR3; 0-2 microg ml(-1)), or IGF-I signal transduction pathway inhibitors (phosphatidylinositol (PI)-3-kinase: 100 nmol Wortmannin l(-1) and 10 micromol LY 294002 l(-1); MAP kinase: 50 micromol PD 98059 l(-1)) to investigate their effects on IGF-I (100 ng ml(-1)) stimulated progesterone secretion. Pig luteal cells displayed dose dependent responses to IGF-I and long R(3)-IGF-I on days 4 and 7 of the oestrous cycle, but not on days 10-16. There was no difference in the ED(50) or V(max) (maximal response) values between IGF-I and long R(3)-IGF-I. Neither anti-IGF-I nor anti-IGF-IR had significant effects on progesterone secretion, at any dose or day. Wortmannin and LY 294002 blocked IGF-I stimulated progesterone secretion, but PD 98059 was without effect. Finally, IGF-I (6 microg) infused into the ovary on day 7 in vivo significantly increased progesterone secretion within 45 min of infusion. The conclusions of this study are: (1) IGF-I has steroidogenic actions only on 'young' (days 4-7) pig corpora lutea; (2) endogenous IGF-I and IGFBP are insufficient to modulate progesterone secretion in vitro; and (3) the steroidogenic actions of IGF-I are mediated via PI-3-kinase. PMID- 12578540 TI - Angiogenesis and vascular endothelial growth factor expression in the equine corpus luteum. AB - Precise pharmacological control of the corpus luteum is important in the manipulation of the oestrous cycle in mares. Angiogenesis plays a key role in the growth and regression of the corpus luteum; therefore, influencing the vasculature of the corpus luteum may offer a novel method for controlling its lifespan. In the present study, changes in angiogenesis and vascular expression of endothelial growth factor (VEGF) were evaluated throughout the luteal phase and after PGF(2alpha)-induced luteolysis. Corpora lutea were collected from mares in the early luteal phase (days 3-4), mid-luteal phase (day 10), early regression (day 14), late regression (day 17), and at 12 and 36 h after administration of PGF(2alpha) on day 10 of the oestrous cycle. Immunohistochemistry was used to localize Von Willebrand factor and Ki67 in endothelial and proliferating cells, respectively. VEGF mRNA and protein were localized by in situ hybridization and immunohistochemistry. The proliferation index of endothelial cells was intense in the early luteal phase. The early and mid-luteal phases were characterized by a dense network of capillaries. The microvasculature started to regress by day 14. After administration of PGF(2alpha), vasodilation was observed after 12 h, but after 36 h, luteal degeneration was accompanied by a significant decrease in vascularity. VEGF mRNA and protein were expressed mainly in the luteal cells during the early and mid-luteal phases and expression declined at early regression (day 14). However, immunostaining for VEGF protein was high in late luteal regression (day 17) and 36 h after PGF(2alpha) administration. These findings indicate a close temporal association between VEGF expression and angiogenesis in the equine corpus luteum during its functional lifespan. PMID- 12578541 TI - Establishment of a recombinant expression system for connective tissue growth factor (CTGF) that models CTGF processing in utero. AB - Connective tissue growth factor (CTGF) stimulates cell proliferation, migration, adhesion and extracellular matrix production, and functions in processes such as development, differentiation, angiogenesis, implantation, wound healing and fibrosis. CTGF is a 38 kDa protein that comprises four discrete structural modules (modules 1-4) but is susceptible to limited proteolysis in utero yielding bioactive isoforms that comprise either modules 3 and 4 (16-20 kDa) or module 4 (10 kDa). Here we report the development of a stable cell line, termed DB1, that was generated by transfecting cDNA encoding full-length human CTGF into Chinese hamster ovary cells that were mutant for heparin sulphate and chondroitin sulphate. DB1 cells produced 38 kDa CTGF and low molecular mass CTGFs that had N termini between modules 2 and 3 at Ala(181) (20 kDa), Leu(184) (18 kDa) or Ala(197) (16 kDa) or between modules 3 and 4 at Gly(253) (10 kDa). CTGF was exported from DB1 cells as early as 5 min after synthesis and all isoforms were readily purified from conditioned medium by sequential steps of heparin affinity, cation exchange, and reverse-phase chromatography. The 38 kDa CTGF was faithfully glycosylated and underwent limited proteolysis in the presence of thrombin, kallikrein or uterine fluids, the last of which was antagonized by anti-thrombin III. All CTGF isoforms promoted cell adhesion, mitosis and epithelial transdifferentiation in vitro as well as subcutaneous fibrosis in vivo. The establishment of this recombinant expression system allows for mass-scale production of all previously reported uterine CTGF isoforms, demonstrates that module 4 contains functional domains involved in a broad range of biological activities, and will facilitate studies of CTGF processing in vitro. PMID- 12578542 TI - Genetic divergence in cellular resistance to heat shock in cattle: differences between breeds developed in temperate versus hot climates in responses of preimplantation embryos, reproductive tract tissues and lymphocytes to increased culture temperatures. AB - The detrimental effects of heat stress on fertility in cattle are less pronounced in heat-tolerant breeds. Although these genetic differences reflect differences in thermoregulation, cells from heat-tolerant breeds are less adversely compromised by increased temperature (that is, heat shock) than cells from heat sensitive breeds. Experiments were performed to test the hypothesis that cells and tissues from two thermotolerant breeds (Brahman and Senepol) are better able to survive and function after exposure to increased temperature than cells and tissues from two thermosensitive breeds (Holstein and Angus). Exposure of embryos at>eight-cell stage at day 5 after insemination to heat shock of 41.0 degrees C for 6 h decreased development to the blastocyst stage and the number of cells per embryo. However, the deleterious effect of heat shock on blastocyst formation and the number of cells per embryo was less pronounced for Brahman than for Holstein and Angus breeds. Embryos from Senepol cows had very low development and it was not possible to determine heat shock effects in this breed. In contrast to the sensitivity of embryos to heat shock, there was no effect of a 41.0 degrees C heat shock on [(3)H]leucine incorporation into proteins secreted by oviductal or endometrial explants. Lymphocytes from Brahman and Senepol cows were more resistant to heat-induced apoptosis than lymphocytes from other breeds. Heat shock reduced lymphocyte glutathione content but the magnitude of the decrease was not affected by breed. In conclusion, embryos from Brahman cows are more resistant to heat shock than embryos from Holstein or Angus cows. Genetic differences are also present in thermotolerance for apoptosis response in lymphocytes, with Brahman and Senepol cattle being more resistant to heat shock than Angus and Holstein breeds. It is likely that the evolutionary forces that led to the Brahman and Senepol breeds being adapted to hot climates resulted in the selection of genes controlling resistance to cellular heat shock. PMID- 12578543 TI - Effects of Gram-positive bacterial pathogens in ewes: peptidoglycan as a potential mediator of interruption of early pregnancy. AB - Bacterial cell walls contain peptidoglycan (PTG), which, among other actions, induces fever. The present experiment evaluated the effects of PTG treatment on early pregnancy and blood plasma concentrations of reproductive hormones. Ewes were injected i.v. with saline or 15, 30 or 60 microg kg(-1) sonicated PTG (Streptococcus pyogenes) on day 5 after mating. Each dose of PTG induced fever. Pregnancy rate at day 25 was not related to incidence of fever but tended to differ among treatments (control, 100%; low, 100%; medium, 67%; high, 60%; P < 0.08). Combined pregnancy rate in ewes from control and low dose groups (100%) was greater than that in ewes from medium and high dose groups (64%, P < 0.01). Ewes with high 13,14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM) concentrations had lower pregnancy rates (6 of 10) than those with low concentrations of PGFM (11 of 11; P < 0.05). Mean cortisol concentrations were higher in treated (2.8 +/- 0.28 microg dl(-1)) than in control (1.1 +/- 0.03 microg dl(-1)) ewes (P < 0.01); the pattern of secretion was biphasic and increased in all treated ewes (P < 0.01). Neither means nor profiles of oestradiol differed with treatment. Mean concentrations and the pattern of concentrations of progesterone were reduced in all treated ewes, as indicated by the time by treatment and linear interaction with treatment (1.2 +/- 0.1 versus 1.6 +/- 0.1 ng ml(-1), P < 0.01). Patterns of LH pulses did not differ from 0 to 4 h or 24 to 28 h after treatment; mean plasma LH concentration was lower in ewes treated with 0, 15 or 30 microg PTG kg(-1) than with 60 microg PTG kg(-1) (P < 0.01). Pregnancy status was not related to plasma concentrations or patterns of LH, oestradiol, progesterone or cortisol. Inflammatory mediators, such as PGF(2alpha), may act directly on the embryo or uterus in ewes treated with PTG. PMID- 12578544 TI - Screening for glycosylation changes on recombinant human IgG using lectin methods. AB - Two lectin-binding methods were investigated as possible ways of monitoring the glycosylation of human monoclonal antibodies during their development and production. Carbohydrate composition was assessed in various preparations that were produced in different host cell types, cell sublines or batches of the same cells. The lectin binding was measured with ELISA and surface plasmon resonance (SPR). For comparative purposes, the monosaccharide content of many of the preparations was also measured by high-pressure anion exchange chromatography (HPAEC)/pulsed amperometric detection (PAD). Both lectin methods detected modifications in glycosylation when antibodies were produced in different ways; SPR was more sensitive than ELISA for some lectins and vice versa. Generally, the lectin results agreed with those obtained by the monosaccharide analysis; however, the former were much better for assessing N -acetylneuraminic acid changes. The latter were impossible to assess by HPAEC/PAD because of their low levels. The lectin-based methods also had the advantages that they were quicker to perform and required less expertise and could quickly identify structures that monosaccharide analysis might miss. It is suggested that, in the development of therapeutic proteins, monosaccharide analysis and/or oligosaccharide profiling is initially performed but later routine batches of the glycoprotein are screened with a lectin method. Of the two lectin methods used, SPR is much quicker when performing a screen, whereas ELISA is particularly useful for comparing a particular carbohydrate feature on different samples of the same glycoprotein. PMID- 12578545 TI - Purification of lysozyme from other hen's-egg-white proteins using metal-affinity precipitation. AB - It was found that the presence of 5 mM Cu(2+) caused precipitation of protein present in hen's egg white to a large extent. About 85% of lysozyme activity remained in the supernatant and the enzyme was purified by approx. 13-fold. A further gel-filtration step on Sephadex G-75 resulted in an overall yield of 80% for the enzyme with 655-fold purification, and showed a single band on SDS/PAGE. PMID- 12578546 TI - Cissus sicyoides (princess vine) in the long-term treatment of streptozotocin diabetic rats. AB - Leaf decoctions of Cissus sicyoides (princess vine) are taken widely as a popular remedy for diabetes mellitus in Brazil, where its common name is 'vegetal insulin'. However, there have been practically no attempts so far to determine scientifically whether it has anti-diabetic effects and we decided to administer leaf decoctions, over extended periods, to normal and streptozotocin-diabetic rats, and investigate the effects of this treatment on the physiological and metabolic parameters that are altered in diabetic animals. The experimental model adopted was shown to be appropriate by running a parallel treatment with insulin, which led to expected improvements in several abnormal parameter values. The decoction treatment significantly reduced the intake of both food and fluid and the volume of urine excreted, as well as the levels of blood glucose, urinary glucose and urinary urea, in comparison with controls. Lipid metabolism was not affected by the treatment; nor was the level of hepatic glycogen in diabetic animals, which indicated that the mechanism responsible for the improvement in carbohydrate metabolism, observed in animals treated with the decoction, could not involve inhibition of glycogenolysis and/or stimulation of glycogenesis. The fact that normal animals treated with C. sicyoides exhibited no changes in any of the measured parameters suggests that its mode of action in diabetic animals does not resemble those of sulphonylurea or insulin. It may, however, act in a similar way to biguanide, via inhibition of gluconeogenesis. PMID- 12578547 TI - A new antibiotic, TH818, and its properties. AB - Antibiotic TH818 was isolated and purified from a culture broth of Streptomyces fulvoviolaceus 818 by extraction and reversed-phase liquid chromatography. The antibiotic TH818 possesses high molecular mass and unique structure, constructed from carbohydrates, amino acids and lipids. The hydrolysis experiments showed the presence of six fatty acids (hexadecanoic, hexadecenoic, octadecanoic, iso nonadecanoic, docosanoic and pentacosanoic acids), four sugars (glucose, galactose, N -acetylglucosamine and N -acetylgalactosamine), and seven amino acids (threonine, alanine, glutamic acid, glycine, serine and two unidentified). TH818 has a broad spectrum of anti-microbial activity against Gram-positive and Gram-negative bacteria, yeasts and fungi. PMID- 12578548 TI - A simplified method for identification of human cardiac myosin heavy-chain isoforms. AB - Cardiac myosin is a central participant in the cross-bridge cycling that mediates myocyte contraction and consists of multiple subunits that mediate both hydrolysis of ATP and mechanical production of contractile force Two isoforms of myosin heavy chain (MHC- alpha and MHC- beta ) are known to exist in mammalian cardiac tissue, and it is within this myosin subunit that ATPase activity resides. These isoforms differ by less than 0.2% in total molecular mass and amino acid sequence, but, strikingly, influence the rate and efficiency of energy utilization for generation of contractile force. Changes in the MHC- alpha /MHC- beta ratio has been classically viewed as an adaptation of a failing myocyte in both animal models and humans; however, their measurement has traditionally required specialized preparations and materials for sufficient resolution. Here we describe a greatly simplified method for routine assessments of myosin isoform composition in human cardiac tissues. The primary advantages of our approach include higher throughput and reduced supply costs with no apparent loss of statistical power, reproducibility or achieved results. Use of this more convenient method may provide enhanced access to an otherwise specialized technique and could provide additional opportunity for investigation of cardiac myocyte adaptive changes. PMID- 12578549 TI - Mammalian cell production and purification of progenipoietin, a dual-agonist chimaeric haematopoietic growth factor. AB - One member of the progenipoietin (ProGP) family of engineered proteins, ProGP-2, is a chimaeric dual cytokine receptor agonist, expressed in mammalian cells, that stimulates both human fetal liver tyrosine kinase-3 (Flt3) and the granulocyte colony-stimulating-factor (G-CSF) receptor. The production of ProGP-2 on a small and large scale using either anti-(Flt3 ligand) antibody-affinity chromatography, or a combination of (NH4)2SO4 fractionation, anion-exchange chromatography, hydrophobic-interaction chromatography and preparative reverse-phase chromatography is described. ProGP-2 was produced in hollow-fibre reactors containing stably transfected NS0 cells. The productivity of ProGP-2 was initially high, but was found to decrease 3-4-fold over time. When the yield of ProGP-2 decreased, the combination of three conventional chromatography steps was required to meet protein purity similar to that achieved by the anti-(Flt3 ligand) chromatography method. In addition, a protease activity was observed in conditioned media from the hollow-fibre reactors that resulted in increased degradation of ProGP-2 that was removed by hydrophobic-interaction chromatography at higher pH. Together the results demonstrated a method for production and purification of ProGP-2 for additional studies on its haematopoietic activity. PMID- 12578550 TI - Inhibitor studies of isopentenyl pyrophosphate biosynthesis in suspension cultures of the yew Taxus chinensis var. mairei. AB - Some metabolic inhibitors, mevastatin (MVS), chlorocholine chloride (CCC), sodium pyrophosphate (NaPP) and D,L-glyceraldehyde (DLG), were respectively added into the suspension cultures of the yew Taxus chinensis var. mairei at the late phase of cell growth to study isopentenyl pyrophosphate (IPP) biosynthesis. The content of total taxanes decreased in the cases of MVS, NaPP and DLG addition, regardless of the inhibitor concentration, whereas it increased in the case of CCC addition. It was thus presumed that a mevalonate pathway might exist in IPP biosynthesis at the late growth phase of Taxus cells and that some IPP might transfer from the cytoplasm to the plastids. This presumption was also confirmed by analysing the effects of the inhibitors on taxane content. It was further demonstrated that IPP in the biosynthesis of taxol, baccatin III and 10-deacetylbaccatin III might transfer from the cytoplasm to the plastids, whereas the translocation of IPP might be not involved in cephalomannine biosynthesis. Furthermore, in combination with the results of previous studies, it is very probable that different IPP biosynthesis pathways exist during different growth phases in Taxus cells. PMID- 12578551 TI - Measurement of NADH concentration in normal and malignant human tissues from breast and oral cavity. AB - We report measurements of the concentration of NADH from malignant and non malignant sites of tissues resected from patients suffering from cancer of the breast or oral cavity. The concentration of NADH was found to be significantly higher in malignant breast-tissue sites as compared with normal breast tissue, whereas the opposite result was obtained with malignant and normal tissues from the oral cavity. These results confirm the inferences made previously, from spectroscopic studies, and are consistent with other biochemical measurements reported in breast and oral cavity tissues. PMID- 12578552 TI - Process characterization of the chromatographic steps in the purification process of a recombinant Escherichia coli-expressed protein. AB - Process development and characterization studies were performed for the chromatographic steps in the purification process of a recombinant Escherichia coli -expressed protein product candidate. The objective of this work was to develop a robust and efficient purification process that would generate material of adequate purity and quantity. A resin screening procedure was developed to aid in picking out the optimal resin for each of the chromatographic columns. It was found that, as a result of resin screening, it was possible to come up with a process with only two column-chromatographic steps. The resulting process used a sulphopropyl (SP) and a quaternary amino (Q) column with intermittent ultrafiltration steps for purification. Effects of different process parameters such as the gradient slope, pH, flow velocity and protein loading on the column performance were evaluated. Buffer pH for the SP column, and buffer pH, gradient slope, protein loading and flow velocity for the Q column, were identified as parameters that could have a significant impact on the performance of the chromatographic step and would require further characterization to improve the robustness of the process. Further process characterization led to the findings that the gradient slope, load pH and buffer pH of the Q column have a significant impact on column performance (>15% change in step yield). All other parameters under consideration did not have any significant impact on pool quality (>10% change in pool purity for the SP column and >5% for the Q column). On the basis of small-scale studies, optimum operating conditions were chosen and the purification process was successfully scaled-up to a large-scale robust process with step yields and product quality that were better than those at the small scale. PMID- 12578553 TI - Lipase assays for conventional and molecular screening: an overview. AB - Lipases are versatile biocatalysts that can perform innumerable different reactions. Their enantio-, chemo- and stereo-selective nature makes them an important tool in the area of organic synthesis. Unlike other hydrolases that work in aqueous phase, lipases are unique as they act at the oil/water interface. Besides being lipolytic, lipases also possess esterolytic activity and thus have a wide substrate range. Hence, the lipase assay protocols hold a significant position in the field of lipase research. Lipase activity can be estimated using a wide range of assay protocols that differ in terms of their basic principle, substrate selectivity, sensitivity and applicability. As the value of these enzymes continues to grow and new markets are exploited, development of new or improved enzymes will be a key element in the emerging realm of biotechnology. Hence, development of faster and simpler protocols incorporating newer and more specific substrates is the need of the hour. In this endeavour, methods that could be adopted for molecular screening occupy an important position. Here, an overview of the lipase assay protocols is presented with emphasis on the assays that can be adopted for the molecular screening of these biocatalysts. PMID- 12578554 TI - Preparation of small unilamellar vesicles (SUV) and biophysical characterization of their complexes with poly-l-lysine-condensed plasmid DNA. AB - Liposomes have numerous applications in the (bio)pharmaceutical industries as agents in the synthesis of new biomaterials for use in areas including gene delivery. There is currently a need to establish efficient scaleable methods for the manufacture of liposomes, and in the present paper we describe the operation of a new high-velocity jet homogenizer for downsizing of multilamellar large vesicles to produce small unilamellar vesicles (SUV). Measurements of size distribution of SUVs are presented and compared with mathematical simulations based on the solution of a population balance equation combined with computational-fluid-dynamics analysis of flow in the homogenizer. Anionic SUVs are produced by the new method and incubated with poly-L-lysine (PLL)-condensed plasmid DNA (pDNA) to generate complexes under different physico-chemical conditions. The colloidal properties of the resulting complexes, including their size and charge, are measured using a Zetasizer and the encapsulation efficiency is obtained experimentally using a Pico Green assay. The results show that between 85 and 95% of the PLL-pDNA condensed plasmids were encapsulated by the liposomes, the smaller liposomes being more effective in encapsulating the complexes. PMID- 12578555 TI - Effects of fermentation strategy on the characteristics of plasmid DNA production. AB - The synthesis of supercoiled plasmid DNA (SC-pDNA) for therapeutic use will involve large-scale production in bioreactors. The success of these fermentations will be dependent on the interactions between the host organism, the recombinant plasmid vector and the growth environment. In the present study, the recombinant host, Escherichia coli DH5 alpha bearing the recombinant plasmid pSV beta, was grown in shake flasks, batch and exponentially fed-batch bioreactors. Specific and volumetric pDNA yields were increased 8- and 25-fold respectively using exponentially fed-batch cultures in comparison with shake-flask cultures. The percentage of SC-pDNA as a proportion of total plasmid DNA decreased over time in batch cultures, but remained relatively constant during fed-batch cultures. The relative merits of different modes of fermentation and their effects on the quality of alkaline lysate extracts of pDNA with respect to genomic contamination and the percentage of SC-pDNA are discussed. PMID- 12578556 TI - Elicitation of plants and microbial cell systems. AB - Plants show physiological and morphological responses to a range of physical and chemical factors known as 'elicitors'. These responses have been considered as defence reactions 'elicited' by the plants' biochemical factory to ensure their survival, persistence and competitiveness. Recently examples have been cited of elicitation in some fungal and bacterial cultures. Through a chronological survey, this Review considers examples of elicitors and elicitation and describes suggested mechanisms of elicitation in plants and microbial cell cultures. The majority of research in this field has been carried out on the plant systems using complex (undefined) biotic elicitors. Carbohydrates are the main class of compounds used as defined elicitors. This Review focuses on carbohydrates as compounds initiating a defence response in cell cultures. Physiological changes brought about on the plant and microbial cultures include expression of novel metabolites and overproduction of already known products. Recent reports confirming elicitation in microbial cultures are of potential importance, as the relative ease of fermentation and scale-up could open an opportunity for the introduction of useful novel metabolites as well as enhancement of commercially useful bioproducts. In this context, a sound knowledge of the elicitor molecules' structure-function relationships and mechanisms of elicitation is essential. PMID- 12578557 TI - Novel insights into how purines regulate pituitary cell function. AB - Purine nucleosides and nucleotides are widely distributed substances that exhibit a diverse range of effects in a number of tissues, acting as important extracellular signalling molecules in addition to their more established roles in cellular metabolism. They mediate their effects via activation of distinct cell surface receptors, termed adenosine (or P1) and P2 purinergic receptors. Although roles for adenosine and adenine nucleotides have been described previously in the pituitary gland, the distribution of the receptor subtypes and the effects of their activation on pituitary function are not well defined. Recent evidence, however, has emerged to describe a complex signalling system for purines in the pituitary gland. Data from a variety of studies have shown that the expression pattern, number and affinity of adenosine and/or P2 receptors may be cell-type specific and that non-endocrine in addition to endocrine cells elaborate these receptors. These variations, along with the diverse range of signalling pathways activated, dictate the response of individual cell types to extracellular purines, with roles now emerging for these substances in the regulation of hormone release, pituitary cell proliferation and cytokine/growth factor expression. In this review, we discuss these advances and examine some implications for pituitary growth control and the response of the hypothalamic pituitary-adrenal axis to stress and inflammation. PMID- 12578559 TI - Transport activity of chimaeric AE2-AE3 chloride/bicarbonate anion exchange proteins. AB - Chloride/bicarbonate anion exchangers (AEs), found in the plasma membrane of most mammalian cells, are involved in pH regulation and bicarbonate metabolism. Although AE2 and AE3 are highly similar in sequence, AE2-transport activity was 10-fold higher than AE3 (41 versus 4 mM x min(-1) respectively), when expressed by transient transfection of HEK-293 cells. AE2-AE3 chimaeras were constructed to define the region responsible for differences in transport activity. The level of AE2 expression was approx. 30% higher than that of AE3. Processing to the cell surface, studied by chemical labelling and confocal microscopy, showed that AE2 is processed to the cell surface approx. 8-fold more efficiently than AE3. The efficiency of cell-surface processing was dependent on the cytoplasmic domain, since the AE2 domain conferred efficient processing upon the AE3 membrane domain, with a predominant role for amino acids 322-677 of AE2. AE2 that was expressed in HEK-293 cells was glycosylated, but little of AE3 was. However, AE2 expressed in the presence of the glycosylation inhibitor, tunicamycin, was not glycosylated, yet retained 85 +/- 8% of anion-transport activity. Therefore glycosylation has little, if any, role in the cell-surface processing or activity of AE2 or AE3. We conclude that the low anion-transport activity of AE3 in HEK-293 cells is due to low level processing to the plasma membrane, possibly owing to protein interactions with the AE3 cytoplasmic domain. PMID- 12578560 TI - Intracellular metabolism and bioactivity of quercetin and its in vivo metabolites. AB - Understanding the cellular effects of flavonoid metabolites is important for predicting which dietary flavonoids might be most beneficial in vivo. Here we investigate the bioactivity in dermal fibroblasts of the major reported in vivo metabolites of quercetin, i.e. 3'-O-methyl quercetin, 4'-O-methyl quercetin and quercetin 7-O-beta-D-glucuronide, relative to that of quercetin, in terms of their further metabolism and their resulting cytotoxic and/or cytoprotective effects in the absence and presence of oxidative stress. Uptake experiments indicate that exposure to quercetin led to the generation of two novel cellular metabolites, one characterized as a 2'-glutathionyl quercetin conjugate and another product with similar spectral characteristics but 1 mass unit lower, putatively a quinone/quinone methide. A similar product was identified in cells exposed to 3'-O-methyl quercetin, but not in the lysates of those exposed to its 4'-O-methyl counterpart, suggesting that its formation is related to oxidative metabolism. There was no uptake or metabolism of quercetin 7-O-beta-D-glucuronide by fibroblasts. Formation of oxidative metabolites may explain the observed concentration-dependent toxicity of quercetin and 3'-O-methyl quercetin, whereas the formation of a 2'-glutathionyl quercetin conjugate is interpreted as a detoxification step. Both O -methylated metabolites conferred less protection than quercetin against peroxide-induced damage, and quercetin glucuronide was ineffective. The ability to modulate cellular toxicity paralleled the ability of the compounds to decrease the level of peroxide-induced caspase-3 activation. Our data suggest that the actions of quercetin and its metabolites in vivo are mediated by intracellular metabolites. PMID- 12578561 TI - Specificity of S'1 and S'2 subsites of human tissue kallikrein using the reactive centre loop of kallistatin: the importance of P'1 and P'2 positions in design of inhibitors. AB - We have demonstrated that the S'(1) and S'(2) subsites of human tissue kallikrein (hK1) play determinant roles in the recognition and hydrolysis of substrates. The presence of serine at position P'(1) and arginine at P'(2) resulted in the best substrate, Abz-Ala-Ile-Lys-Phe-Phe-Ser-Arg-Gln-EDDnp, which was derived from the kallistatin reactive-centre loop sequence and quencher groups o-aminobenzoic acid (Abz) and N-(2,4-dinitrophenyl)ethylenediamine (EDDnp). Serine and arginine are also the residues at positions P'(1) and P'(2) in human kininogen, from which hK1 releases Lys-bradykinin. Several peptide analogues of Abz-Ala-Ile-Lys-Phe-Phe-Ser Arg-Gln-EDDnp, in which the Ser and Arg residues were substituted with various other amino acids, were synthesized and tested as substrates. Most of them were hydrolysed slowly, although they showed significant binding to hK1, as demonstrated by their competitive inhibition constants (K(i)). Using this information, six peptides were designed, synthesized and assayed as inhibitors of hK1. Abz-Lys-Phe-Phe-Pro-Arg-Gln-EDDnp, Abz-Lys-Phe-Arg-Pro-Arg-Gln-EDDnp and acetyl-Lys-Phe-Phe-Pro-Leu-Glu-NH(2) inhibited hK1 in the range 20-30 nM (letters in italics denote the D-form of the amino acid). The peptide acetyl-Lys-Phe-Phe Pro-Leu-Glu-NH(2) was a weak inhibitor for other serine proteases, as indicated by the higher K (i) values compared with hK1, but this peptide was a potent inhibitor of human plasma kallikrein, which has a K (i) value of 8 nM. This result was surprising, since this enzyme is known to be a restricted arginyl hydrolase. In conclusion, acetyl-Lys-Phe-Phe-Pro-Leu-Glu-NH(2) can be used as a leader compound to design specific inhibitors for hK1, plasma kallikrein, or for both at same time, if the inhibition of kinin release is the main goal. PMID- 12578558 TI - Mammalian molybdo-flavoenzymes, an expanding family of proteins: structure, genetics, regulation, function and pathophysiology. AB - The molybdo-flavoenzymes are structurally related proteins that require a molybdopterin cofactor and FAD for their catalytic activity. In mammals, four enzymes are known: xanthine oxidoreductase, aldehyde oxidase and two recently described mouse proteins known as aldehyde oxidase homologue 1 and aldehyde oxidase homologue 2. The present review article summarizes current knowledge on the structure, enzymology, genetics, regulation and pathophysiology of mammalian molybdo-flavoenzymes. Molybdo-flavoenzymes are structurally complex oxidoreductases with an equally complex mechanism of catalysis. Our knowledge has greatly increased due to the recent crystallization of two xanthine oxidoreductases and the determination of the amino acid sequences of many members of the family. The evolution of molybdo-flavoenzymes can now be traced, given the availability of the structures of the corresponding genes in many organisms. The genes coding for molybdo-flavoenzymes are expressed in a cell-specific fashion and are controlled by endogenous and exogenous stimuli. The recent cloning of the genes involved in the biosynthesis of the molybdenum cofactor has increased our knowledge on the assembly of the apo-forms of molybdo-flavoproteins into the corresponding holo-forms. Xanthine oxidoreductase is the key enzyme in the catabolism of purines, although recent data suggest that the physiological function of this enzyme is more complex than previously assumed. The enzyme has been implicated in such diverse pathological situations as organ ischaemia, inflammation and infection. At present, very little is known about the pathophysiological relevance of aldehyde oxidase, aldehyde oxidase homologue 1 and aldehyde oxidase homologue 2, which do not as yet have an accepted endogenous substrate. PMID- 12578562 TI - The mitochondria-associated endoplasmic-reticulum subcompartment (MAM fraction) of rat liver contains highly active sphingolipid-specific glycosyltransferases. AB - Although most glycosphingolipids (GSLs) are thought to be located in the outer leaflet of the plasma membrane, recent evidence indicates that GSLs and their precursor, ceramide, are also associated with intracellular organelles and, particularly, mitochondria. GSL biosynthesis starts with the formation of ceramide in the endoplasmic reticulum (ER), which is transported by controversial mechanisms to the Golgi apparatus, where stepwise addition of monosaccharides on to ceramides takes place. We now report the presence of GSL-biosynthetic enzymes in a subcompartment of the ER previously characterized and termed 'mitochondria associated membrane' (MAM). MAM is a membrane bridge between the ER and mitochondria that is involved in the biosynthesis and trafficking of phospholipids between the two organelles. Using exogenous acceptors coated on silica gel, we demonstrate the presence of ceramide glucosyltransferase (Cer-Glc T), glucosylceramide galactosyltransferase and sialyltransferase (SAT) activities in the MAM. Estimation of the marker-enzyme activities showed that glycosyltransferase activities could not be ascribed to cross-contamination of MAM by Golgi membranes. Cer-Glc-T was found to have a marked preference for ceramide bearing phytosphingosine as sphingoid base. SAT activities in MAM led to the synthesis of G(M3) ganglioside and small amounts of G(D3). G(M1) was also synthesized along with G(M3) upon incubation of the fraction with exogenous unlabelled G(M3), underlying the presence of other sphingolipid-specific glycosyltransferases in MAM. On the basis of our results, we propose MAM as a privileged compartment in providing GSLs for mitochondria. PMID- 12578564 TI - [Phenotypical and Biological Characteristics of Human Fetal Marrow and Liver Mesenchymal Stem Cells] AB - There has been an increasing interest in recent years in the stromal cell system. The stroma is a complex tissue, composed of a number vascular and connective tissue cell types including endothelial cells, adipocytes, smooth muscle cells, osteogenic cells, and stromal cells. The marrow mesenchymal stem cells are capable of self-renewal and differentiate into various connective tissue lineages including bone, cartilage, tendon, muscle, fat, and marrow stroma. Recently, techniques for the isolation of adult bone marrow-derived human and animal mesenchymal stem cells have been described, as well as the methods to directing their differentiation into osteogenic, chondrogenic, tendogenic, myogenic, adipogenic, and marrow stromal lineages. But there is no report about the fetal bone marrow- and liver-derived mesenchymal stem cells. Are they the same or not? In our assay, human fetal mesenchymal stem cells from 4 - 5 months old human fetal bone marrow and liver low-density mononuclear cells were cultured, and the cell cycle, immunophenotype and ex vivo expansion properties were studied. Results showed that the mesenchymal stem cells from fetal liver and fetal bone marrow were similar in morphology, growth character and immuno-phenotypes, but the liver mesenchymal cells manifested higher ability to support hematopoiesis than the marrow-derived mesenchymal cells. This study demonstrates that adherent fetal marrow- and liver-derived cells cultured in the absence of differentiation stimulus gave rise to a population of cells with phenotypical features of mesenchymal stem cells, and should be enough to sustain a steady supply of low differentiated cells upon proliferation. PMID- 12578563 TI - Prion peptide induces neuronal cell death through a pathway involving glycogen synthase kinase 3. AB - Prion diseases are characterized by neuronal cell death, glial proliferation and deposition of prion peptide aggregates. An abnormal misfolded isoform of the prion protein (PrP) is considered to be responsible for this neurodegeneration. The PrP 106-126, a synthetic peptide obtained from the amyloidogenic region of the PrP, constitutes a model system to study prion-induced neurodegeneration as it retains the ability to trigger cell death in neuronal cultures. In the present study, we show that the addition of this prion peptide to cultured neurons increases the activity of glycogen synthase kinase 3 (GSK-3), which is accompanied by the enhanced phosphorylation of some microtubule-associated proteins including tau and microtubule-associated protein 2. Prion peptide treated neurons become progressively atrophic, and die ultimately. Both lithium and insulin, which inhibit GSK-3 activity, significantly decrease prion peptide induced cell death both in primary neuronal cultures and in neuroblastoma cells. Finally, the overexpression of a dominant-negative mutant of GSK-3 in transfected neuroblastoma cells efficiently prevents prion peptide-induced cell death. These results are consistent with the view that the activation of GSK-3 is a crucial mediator of prion peptide-induced neurodegeneration. PMID- 12578565 TI - [Antisense Oligodeoxynucleotide Directed to NF-kappaB-RelA Down-Regulates bcl x(L) mRNA in Drug-Resistant Leukemia Cell Line HL-60/E6] AB - In order to explore the effect of nuclear factor-kappa B (NF-kappaB) on bcl-x gene transcrtiption in drug-resistant leukemia cell line HL-60/E6, first of all, drug-resistant subline HL-60/E6 was derived by intermittently exposing HL-60 cells to 6 ng/ml epirubicin, and then bcl-x(L) mRNA levels were detected by RT PCR after exposing HL-60/E6 cells to 5 micro mol/L AS-PS-ODN-RelA at different times. Morever, indirect immuno-fluorescence and FCM were used to demonstrate the location of NF-kappaB-RelA in HL-60/E6 cells and the efficiency of liposome mediated ODN transfection. The results showed that RelA kept active and located at the nuclei of HL-60/E6 cells, and the efficiency of liposome-mediated ODN transfection was significantly higher than that of null ODN (P < 0.01 in 4, 6 and 12 h). Exposure of HL-60/E6 cells to 5 micro mol/L AS-PS-ODN-RelA led to a maximal 40% decline of bcl-x(L) mRNA levels. No significant change of bcl-x(L) mRNA level occurred in control group. It was concluded that NF-kappaB was involved in regulating bcl-x transcription. Suppressing NF-kappaB-RelA by antisense technology may down-regulate level of bcl-x(L) mRNA. PMID- 12578566 TI - [Clinic Significance of Expression of bcl-2 and bax Gene in Patients with Acute Leukemia and its Relationship with mdr-1 Gene Expression] AB - It is generally accepted that the inhibition of apoptosis is one of the mechanism of drug resistance to tumor. Members of the bcl-2 gene family are the most important regulators in apoptosis. The purpose of this study is to evaluate the value of expression of bcl-2 and bax gene in predicting the prognosis of acute leukemia patients, and to explore the relationship between bcl-2 and bax expression and drug resistance. Seventy patients with acute leukemia entered this study. Expressions of bcl-2, bax and mdr-1 gene were measured by RT-PCR method and FCM. The result showed that: bcl-2 had been widely detected in specimens of blood or bone marrow from acute leukemia patients, the expression levels were much higher than those in normal control (1.46 vs 0.71, P < 0.05), bax expression levels and bax/bcl-2 ratio in patients had no significant difference with the control. No relationships were found between the expression levels of bcl-2 and bax and AL patients' age, sex, platelet counts, hemoglobin levels, percentage of marrow blasts, FAB classification, and S + G(2)M%. Both Bcl-2 protein expression (34.6% vs 69.2%, P < 0.03) and bax/bcl-2 mRNA ratio (37.1% vs 82.9%, P < 0.01) were associated with response to therapy and CR rate, bax/bcl-2 ratio also influences the overall survival time. There was no relationship between bcl-2 and bax expression levels and mdr-1 expression levels. PMID- 12578567 TI - [Study of Telomerase Activity in Bone Marrow Cells from Patients with Myelodysplastic Syndromes] AB - To study the telomerase activity in the bone marrow MNCs from patients with myelodysplastic syndromes in comparison with that in normal individuals and acute leukemia patients. The intracellular telomerase activity was semi-quantitatively examined by PCR-ELISA assay in the marrow cells of 20 normal individuals, 21 cases of myelodysplastic syndromes (MDS) and 32 cases of acute leukemia. Telomerase activity in normal marrow cells was 0 - 0.30 U, with a mean level of (0.11 +/- 0.08) U, in which 3 cases were considered positive according to the standard set by the Kit. In 32 acute leukemia patients, the mean level of telomerase activity was (0.42 +/- 0.26) U (ranged 0 - 0.96 U) with a positive rate of 78.1%, showing a significantly higher activity in acute leukemia (P < 0.01). Moderate telomerase activity was detected in 21 cases of MDS, with a mean level of (0.27 +/- 0.19) U (0 - 0.97 U) from which the positive rate was 66.7%. This value was significantly higher than that in the normal BM (P < 0.05). Moreover, a significantly higher telomerase activity was shown in the high-risk group of MDS (P < 0.05). Based on the international scoring system evaluating the prognosis of MDS (IPPS), telomerase activity in HIGH subgroup was significantly higher than that in INT-1 and INT-2 subgroup (P < 0.05). The level of telomerase activity was not correlated to the chromosome aberrations. These results show that a borderline telomerase activity could be found in normal bone marrow cells. Telomerase activity was markedly higher in acute leukemia. BM of MDS patients demonstrated a moderate telomerase activity. Higher telomerase activity could be found in high-risk group and correlated with poor prognosis. PMID- 12578568 TI - [Analysis of Clinical and Hematologic Features in 22 Patients with Metastatic Carcinoma of Bone Marrow] AB - The clinical and hematologic features in 22 patients with metastatic carcinoma of bone marrow were observed and analyzed. Morphology of bone marrow cells, bone marrow biopsy and other accessory examinations were performed. The primary or cardinal symptoms of metastatic carcinoma of bone marrow included anemia (17 cases, 77.3%), ostealgia (10 cases, 45.5%), fever (8 cases, 36.4%), hemorrhage (4 cases, 18.2%) and complicated hemolytic anemia (4 cases, 18.2%). The primary carcinomas, diagnosed by pathologic and accessory examinations, include gastric carcinoma (6 cases, 27%), lung cancer (3 cases, 13.6%), ovarian cancer (2 cases, 9%), mammary cancer, prostatic carcinoma, osteocarcinoma and metastatic malignant melanoma (1 case, respectively), and unknown primary lesion (7 cases, 31.8%). The hematologic features were decrease of hemoglobin (17 cases, 77.3%) and blood plate count (16 cases, 72.7%), leukocytosis (11 cases, 50%), immature leukocytes (14 cases, 63.6%) and erythrocytes (9 cases, 40.9%) seen on the peripheral blood smear, and reticulocytosis (4 cases, 18.2%). Masses of metastatic carcinoma cells can be frequently seen at two sides and tail of bone marrow smear. Bone marrow biopsy of 8 cases demonstrated the infiltration of carcinoma cells with nest-like distribution in the bone marrow cavity. Examination of MRI in 6 case showed destruction of bone and corpus vertebra and abnormal signal focus. Bone marrow biopsy could contribute to improve the accuracy of diagnosis and determine the origin of primary carcinoma. MRI plays an important role in diagnosis of metastatic carcinoma in bone marrow. PMID- 12578569 TI - [Negative Regulatory Effect of T-Lymphocytes on the Expansion of Human Bone Marrow Hematopoietic Cells In Vitro] AB - In this research, the regulatory effects of T-lymphocytes on the expansion of hematopoietic cells in human bone marrow were studied. Anti-CD3 McAb and anti-CD8 McAb were used to eliminate the T-lymphocytes in bone marrow MNCs. Cell surface antigens were analysed by flow cytometry. Hematopoietic cells expanded with hematopoietic growth factors (HGFs) in a liquid culture system and the number of CD34(+) cell, CFU-GM and CFU-GEMM were determined. After cultured with HGFs for 20 days the total number of cells expanded by 75.8, 79.6, 77.4 and 67.0 folds respectively in three experimental groups (anti-CD3 McAb, anti-CD8 McAb and anti CD3 + anti-CD8 McAb) and control group (MNC of bone marrow). The number of CFU-GM in the four groups were 173.67 +/- 18.90, 165.33 +/- 26.58, 170.33 +/- 21.50 and 79.67 +/- 8.33 respectively. The number of CFU-GEMM in the four groups were 431.33 +/- 34.56, 370.33 +/- 42.10, 386.67 +/- 10.02 and 177.67 +/- 26.86 respectively. There were significant differences in the number of CFU-GM and CFU GEMM between experimental groups and control group. The results showed that the T lymphocytes in bone marrow could inhibit the expansion of hematopoietic cells in vitro and the formation of CFU-GM and CFU-GEMM. The regulatory mechanism was to be explored. PMID- 12578570 TI - [Recombinant Human Interleukin 11 (Mega) Promotes Thrombopoiesis in Cancer Patients with Chemotherapy-Induced Myelosuppression] AB - A randomized, selfcross-over and placebo-controlled clinical trial has been taken to evaluated the curative efficacy of rhIL-11 (Mega) for thrombocytopenia in 29 cancer patients with severe myelosuppression induced by chemotherapy. Twenty-five micro g/kg of Mega or placebo was administered subcutaneously once daily starting 24 hours after the completion of chemotherapy, and continuing for 7 to 14 days or until the platelet count reached 300 x 10(9)/L. The results from those in 118 cases performed phase II clinical trial, showed that there were 29 cases with platelet count less than 50 x 10(9)/L in placebo cycle, but there was only 1 case in Mega cycle. The percentage of the patients with platelet count less than 50 x 10(9)/L in placebo cycle of placebo + Mega group was higher than that of Mega + placebo group. The nadir and platelet counts on day 21 after chemotherapy in Mega cycle were 2.04 and 1.43 times more than those in placebo cycle, respectively. The data show that Mega had significant thrombopoietic activity with a long lasting oction for the patients experienced severe myelosuppression. It significantly increases the likelyhood of avoiding thrombocytopenia in cancer patients undergoing chemotherapy and shortens the duration of thrombocytopenia. PMID- 12578571 TI - [Metabolism, Distribution and Excretion of Recombinant Human Thrombopoietin in Mice] AB - The metabolism, distribution and excretion profiles of recombinant human thrombopoietin (rhTPO) in mice were studied by means of (125)I-labeled rhTPO ((125)I-rhTPO) combined with size exclusive high performance liquid chromatography (SHPLC) or trichloroacetic acid (TCA) precipitation analysis. (125)I-rhTPO was prepared by iodogen method. Purification was performed on Sephacryl S-200 HR gel. Radioactive-purity of (125)I-rhTPO identified by SHPLC was (96.9 +/- 1.5)% (n = 3). The proliferation effect of TPO dependent cell line (TD-3) and the increase of peripheral platelet counts in mouse by (125)I-rhTPO demonstrated that (125)I-labeled protein maintained the biological activities of TPO both in vitro and in vivo. SHPLC analysis of serum and urine samples taken after sc 1 micro g/mouse (345 kBq/mouse) of (125)I-rhTPO revealed that there were two lower molecular weight (125)I-degradation metabolites ((125)I-MI and (125)I MII) other than parent molecule. (125)I-MI was mainly found in urine, and (125)I MII was detected both in serum and in urine. The maximal concentration of (125)I rhTPO was reached at 2 hours after injection. The terminal half-life was 10.8 hours, which was much longer than those of other peptides. TCA precipitable radioactivity in tissue showed that the radioactivity in bone marrow was rather high. The highest level was found in urinary system. Levels in adrenals, lymph nodes, and fat were near to that in serum. Lowest was found in brain. The main excretion route was urinary system and (98 +/- 5.6)% of (125)I-rhTPO was excreted within 72 hours after dosing. PMID- 12578572 TI - [Internal Ribosomal Entry Site-Mediated Expression of Human Multidrug Resistance 1 Gene in Human CD34(+) Cells] AB - In order to reach the purpose of co-transferring double drug resistance genes into human CD34(+) progenitor cells to broaden the spectrum of drug resistance, the expression efficiency of human multidrug resistance 1 (MDR1) gene mediated by the internal ribosomal entry site (IRES) was investigated. Two retroviral vectors were transferred into packaging cells. One is pSF-DIM containing double drug resistance genes, in which the translation of MDR1 gene was controlled under an IRES from encephalomyocarditis virus. The other is pSF-MDR1 which only contains MDR1 gene controlled under the same promoter of pSF-DIM. The amphotropic retroviral packaging cells PA317/pSF-DIM and PA317/pSF-MDR1 were obtained with titer of 8 x 10(4) and 1.3 x 10(5) cfu/ml respectively. Human cord blood CD34(+) cells were transduced by supernatant infection. Expression of P-gp was detected by flow cytometry. Compared with the untransduced group, the expression of P-gp in pSF-DIM transduced group and pSF-MDR1 transduced group was elevated 10.92% and 28.82% respectively. However, the expression of P-gp in pSF-MDR1 transduced group was higher than that in pSF-DIM transduced group. The result suggests that MDR1 gene can express in the human progenitor cells under control of IRES. It laid the foundation of subsequence research. The reason on the difference in MDR1 gene expression efficiency between pSF-MDR1 transduced group and pSF-DIM transduced group need further research. PMID- 12578573 TI - [The Prediction of the Molecular Biology Characteristics of Recombinant Fusion Exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40] AB - In order to confirm the reasonability of designed recombinant exotoxin B7-1 Linker-PE40 and B7-2-Linker-PE40, their molecular biology characteristics, such as flexibility, antigenicity, hydrophilicity, epitope and secondary structure, were predicted by using a computer software GOLDKEY. It had been found that the recombinant fusion exotoxin had kept the epitope characterstics of B7-1, B7-2 and PE40, and had not got new epitope, and the antigenicity in flexible linker was extxemely low. The linker inserted in the recombinant fusion exotoxin had low epitope, low antigenicity and high flexibility. Compared to B7-1, B7-2 and PE40, there are several amino acid residues changes in B7-1-Linker-PE40 and B7-2-Linker PE40, respectively, which might have some effect on secondary structure of the recombinant fusion exotoxins. Western blot analysis revealed that both B7-1 Linker-PE40 and B7-2-Linker-PE40 could bind specifically with antibodies against B7-1, B7-2 and PE40, respectively. The result of Western blot was consistant with the computer prediction that the recombinant proteins retain the antigenicity and spacial structure of B7 and PE40. It is suggested that both fusion proteins designed and constructed were resonable and computer analysis would be helpful for us to study the biological activity of the recombinant fusion exotoxin B7-1 Linker-PE40 and B7-2-Linker-PE40 and construct other recombinant proteins further. PMID- 12578574 TI - [The Antitumor Activity of G-CSF-Mobilized Peripheral Blood Mononuclear Cells Activated by IL-2 Alone or in Combination with IL-12] AB - This study was undertaken to investigate the antitumor activity of rhG-CSF mobilized blood mononuclear cells (G-PBMC) from normal donor activated by rhIL-2 alone or in combination with rhIL-12 in vitro. LDH release assay was used to determine the cytotoxicity activity of G-PBMC against K562 (NK-sensitive) and Raji cells (NK-resistant). The phenotype of the activated G-PBMC was assayed by flow cytometry. Results suggested that the cytotoxicity of the fresh G-PBMC was low before IL-2/IL-12 stimulation. G-PBMC developed marked cytotoxicity after 24 hours of IL-2 activation, with more significant increase for further 48 hours induction. When G-PBMCs were exposed to combination of IL-2 and IL-12, the cytotoxicity was significantly enhanced. The proportions of CD3(+) T cells and CD8(+) T cells were increased when G-PBMC incubated with IL-2 for 72 hours. However, CD56(+) cells were significantly elevated when G-PBMC exposed to IL-2 and IL-12 for 7 days. It is concluded that G-PBMC activated by IL-2 had evident antitumor activity, which was further increased when IL-2 in combination with IL 12. These results demonstrate that IL-2 and IL-12 treatment in vitro might promote the graft-versus-leukemia (GVL) response of G-PBMC. PMID- 12578576 TI - [Antitumor Activities of KRN7000 in NS-1 Myeloma-Bearing Mice] AB - To explore the antitumor activities of KRN7000 in a NS-1 myeloma-bearing mice and the underlying mechanisms, cytotoxic activities of the spleen cells treated with KRN7000 in vivo and in vitro were examined with Yac-1 (a NK-sensitive cell line) and NS-1 (a NK-insensitive cell line) as the targeted cells. The life span of NS 1 myeloma-bearing mice was observed after treating with KRN7000 and combined with cyclophosphamide (CTX). Furthermore, toxicities of KRN7000 administration on liver and kidney were evaluated. The results showed that KRN7000 enhanced NK cytotoxic activities of spleen cells. Around 20% of NS-1-bearing mice survived after KRN7000 administration and the survival rate reached up to 80% of NS-1 bearing mice when KRN7000 was used in combination with CTX at a dose of 100 mg/kg (P < 0.005). KRN7000, at a dose of 100 micro g/kg, had no toxic effects on liver and kidney. These findings suggest that KRN7000 might be a promosing agent in tumor immunotherapy. PMID- 12578575 TI - [Effect of Autologous or Allogeneic Anti-CD3 Monoclonal Antibody Activated Killer Cells on Normal Hematopoietic Cells] AB - In order to investigate the effect of autologous and allogeneic anti-CD3 McAb activated killer cells (CD3AK) on normal hematopoietic cells, the immobilized anti-CD3 McAb and low concentration IL-2 were used to activate CD3AK. Flow cytometry was used to assay the phenotype change of CD3AK to analyze the proportional change of CD34(+) cells in normal bone marrow mononuclear cells (BMMNC) cocultured with autologous or allogeneic CD3AK. The effect of CD3AK on normal hematopoietic progenitor cells was also assayed by methylcellulose clonogenic culture of CFU-GM. It was found that 3 - 5 micro g/ml immobilized anti CD3 McAb and 100 U/ml IL-2 could activate CD3AK effectively. There were 99.51% CD3(+) cells in CD3AK groups. When BMMNCs from healthy volunteers were cocultured with allogeneic CD3AK for six hours, the percentage of CD34(+) cells was decreased 32.37%. CD3AK had no significant influence on autologous BMMNC. Allogeneic and autologous CD3AK were cultured with BMMNC from healthy volunteers for six hours, and then CFU-GM was evaluated. Allogeneic CD3AK inhibited 20.44% CFU-GM formation, but autologous CD3AK had no inhibition on CFU-GM. It is concluded that CD3AK has no inhibition to autologous normal hematopoietic progenitor cells after cocultured with them from these results, while allogeneic CD3AK inhibits the normal hematopoietic progenitor cells significantly. PMID- 12578577 TI - [A Experimental Study on Improvement of the Seeding Efficiencies of Infused Donor Hematopoietic Cells in Syngeniec Bone Marrow Transplantation by Aortic Infusion] AB - In order to explore the improvement of seeding efficiencies of infused donor hematopoietic cells to bone marrow in bone marrow transplantation, two recipient groups of syngeneic rat model which received transplanted cells labeled with PKH 26, a red fluorescent membrane dye, by aortic or intravenous administration (2 x 10(7) nucleate cells per recipient rat) respectively, were assayed; at selected times following BMT, partial recipient rat were euthanized and then measured the numbers of PKH-26 labeled cells in recipient rat marrow samples by means of flow cytometry. The results showed that the homing indices of donor hematopoietic cells in aortic group and intravenous group were (14.52 +/- 1.07)% and (10.49 +/- 0.72)% at 30 hours after BMT, respectively (P < 0.05). The results indicated that the number of donor hematopoietic cells localized to recipient bone marrow infused by aortic route is more than that infused by intravenous route. PMID- 12578578 TI - [Study on Relation of Early Immune Reconstitution with Acute Graft-Versus-Host Disease] AB - To analyze the relation of early immune reconstitution with acute graft-versus host disease after allogeneic hematopoietic stem cell transplantation (all-HSCT), the changes of CD3(+), CD4(+), CD8(+), CD25(+) and CD69(+) cells in peripheral blood from 26 patients with hematologic malignancies were assayed by flow cytometry within 2 months after allo-HSCT. All patients achieved hematopoietic reconstitution, and grade I and II - IV GVHD were developed in 9 and 5 patients, respectively. CD25(+) cells were increased in patients aGVHD at week 2 after transplantation and the peak value was appeared at week 3. The increase of CD25(+) cells was preceded the occurence of clinical signs of aGVHD. The maximal levels of CD25(+) cells increase correlated significantly with the severity of aGVHD. The increase of CD25(+) cells was declined along with remission of aGVHD signs. Our results suggest that analyzing immune reconstitution after allo-HSCT could predict occurence of aGVHD, and CD25(+) cell increase prior occurence of aGVHD is predictive marker for aGVHD. PMID- 12578579 TI - [A New Criterion for Donor and Recipient Selection in Hematopoietic Stem Cell Transplantation - the Matching of Three-Dimensional Structure of HLA Molecular Modeling] AB - The purpose of the research is to provide a new standard for matching of HLA three-dimensional structure, and summarize the major permissible mismatch and immunogenic mismatch antigens. The molecular modeling method was used to create HLA molecular structures by Swiss Model Server, and the comparison of the differences among the alleles was done by SPDV software with the function of iterative magic fit. The results were recorded by relative mean square deviation (RMSD, nm). The differences among alleles were scattered below 0.06 nm for HLA-A and -B molecules, and below 0.03 nm for HLA-DRB1 molecules. On the basis of the statistical analysis, when RMSD is greater than 0.04 nm for -A and -B molecules and 0.02 nm for -DRB1 molecules, the difference is meaningful and can be related with graft versus host disease. When RMSD is lower than 0.02 nm for -A and -B molecules and 0.01 nm for -DRB1 molecules, the difference is decided unmeaningful. From the data, the permissible mismatch and immunogenic mismatch alleles within HLA-A, HLA-B and HLA-DRB1 molecules were summarized. Three dimensional structure matching is a new area in the transplantation field, much research should be done in the future. PMID- 12578580 TI - [Typing of HLA-AB by Reverse PCR-SSOP and Clinical Application] AB - A rapid and accurate method of DNA typing for HLA was established to compensate the unsatisfactory serological typing for HLA before transplantation. DNA typing for HLA using by reverse polymerase chain reaction with sequence-spcific oligo probe (reverse PCR-SSOP) could detect HLA-A(*0101 - 8001) and B(*07021 - 8201). The results showed that HLA-AB alleles were successfully analysed in 60 matching subjects and 16 control DNAs from UCLA by reverse PCR-SSOP without false negtive and false positive results. The results were concordance with those of UCLA. The error rate of serological typing was 6.4% for HLA-A and 7.4% for HLA-B. The serological typing missed HLA-A24 and HLA-B46 for two patients with leukemia respectively. Our results suggest that DNA typing for HLA by reverse PCR-SSOP has proved to be a high-resolution, high-specificity, rapid and accurate technique, suitable for clinical application with a greater precision than serological typing. PMID- 12578581 TI - [A Simplified Method for Cryopreservation of Peripheral Blood Stem Cells] AB - To simplify the traditional method for cryopreservation of peripheral blood stem cells(PBSCs) at -196 degrees C with rate-controlled freezing with 10% dimethyl sulfoxide(DMSO), the simplified method was carried out by freezing the cells to 80 degrees C in low-temperature freezer with the combination of 5% DMSO, 3% hydroxyethyl starch(HES) and 4% human serum albumin(HSA) as cryoprotectant. PBSCs were cryopreserved by different methods. Cell viability and recovery rate of mononuclear cells (MNC), CFU-GM and CD34(+) cells were compared. It was observed that the higher MNC and CFU-GM recovery rates were achieved and without agglutination with the simplified method. It was also found with this simplified method, satisfactory recovery rates of CFU-GM and CD34(+) cells could be obtained when PBSCs were preserved at -80 degrees C as long as 24 months. There was no difference observed in parameters of cryopreserved PBSCs thawed at 37 degrees C and 20 degrees C. After the cells being exposed to 5% DMSO at room temperature for 1 hour, the cell viability decreased from 93.2% to 84.5%, however, the CFU-GM recovery rate was not decreased. It is concluded that the simplified cryopreservation technique is better and simpler than the traditional crypreservation method, will be useful for institutions without rate-controlled freezing facility. Moreover, this method diminishes the amount of DMSO infused into patients, thus decreasing its toxicity. PMID- 12578582 TI - [Establishment of Quantitative Assay for Human Erythrocyte Pyrimidine 5' Nucleotidase Content by Double Antibody Sandwich ELISA] AB - For exploring pathogenesis of pyrimidine 5'-nucleotidase (P5'N) deficiency, a quantitative assay method for human erythrocyte pyrimidine 5'-nucleotidase was established. The specific substrate uridine monophosphate (UMP) of P5'N was used as ligand. The UPM-ADH-Sepharose 4B affinity column was prepared. P5'N of human erythrocyte was purified by ammonium sulfate fractionation and precipitation, ion chromatography and affinity chromatography. Rabbit anti-human P5'N antibody was acquired by immunizing rabbits with purified P5'N. Using rabbit anti-human antibody as the coated anti-body and HRP-rabbit anti-human antibody as demonstrated anti-body, the double antitody sandwich ELISA for quantitative assay of human erythrocyte P5'N was established after square rank trial, antigen blocked trial and antigen substituted test. Results showed that the titer of rabbit anti-human erythrocyte was 1:4 and the sensitivity of double antibody sandwich ELISA was 5 ng/ml. Its blocking rate was more than 95% and the rate of substitution less than 30%. The content of P5'N was (71.77 + 10.98) ng/mg NHP in normal human erythrocyte. A new ELISA method for quantitative determination of human erythrocyte P5'N was first established. It not only had high specificity and sensitivity but also could assay the minimun content of P5'N as 5 - 20 ng/ml. It could be a suitable method for large sample in clinics. PMID- 12578583 TI - [Progress in Research on Embryonic Stem Cells] AB - Embryonic stem cells are continuously growing stem cell lines of embryonic origin. The distinguishing features of ES cells are their capacity to be maintained in an undifferentiated state indefinitely in culture and their potential to develop into various kinds of cells in the body. At molecular level, in vivo and in vitro differentiation of ES cells are similar in many ways. To date, ES cells can differentiate in vitro into myocytes and myocardial cells, neural precursor cells, hematopoietic cells and endothelial cells. Recent evidence suggests that hematopoietic stem cells (HSCs) originate from a region within the embryo proper. It is likely that the genesis of HSCs is regulated by embryonic growth factors, which have not been discovered yet. ES cells have provided a convenient and effective experimental approach to define early hematopoietic process within embryos. Research on ES cells has broad application prospect. PMID- 12578584 TI - [Recent Advance in Anti-CD20 Monoclonal Antibody Therapy in B-Cell Lymphoma] AB - Substantial advances in antigen-targeted lymphoma therapy have been achieved in recent years. Monoclonal antibodies targeting B-cell differential antigen CD20 have emerged as promising new treatments. CD20 is a 35 kD non-glycosylated transmembrane phosphoprotein. It is expressed on most mature B-lymphocytes and disappears from the surface of B lineage cells during terminal differentiation into plasma cells. The antigen appears to be involved in the regulation of B-cell development and differentiation and may mediate some of its effects by functioning as a calcium channel. Most importantly, CD20 is expressed on more than 95% of B-cell lymphomas and is not significantly internalized or shed. These features make it an ideal target for monoclonal antibody therapy for B-cell lymphomas. The results of clinical trials have showed that anti-CD20 monoclonal antibodies, which can be utilized in either unmodified form or as carrier for radioisotopes or cytotoxic agents, have significant effects and can be administrated safely with minimum side effects. Many studies have proposed several potential mechanisms to mediate the eradication of tumor cells targeted by anti-CD20 monoclonal antibodies. Ongoing prospective studies will establish some new therapeutic strategies with high anti-lymphoma specificity and low unspecific toxicity. PMID- 12578585 TI - [Telomerase Activity Expressed in Acute Leukemia Cell] AB - To explore the possible linkage between telomerase and acute leukemia, we detected telomerase activity expressed in 3 leukemia cell lines, 22 acute leukemia bone marrow and 6 normal bone marrow with PCR ELISA assay. Results showed that telomerase activities of three leukemia cell lines were positive with the average (1.57 +/- 0.056) U, normal bone marrow samples average was (0.085 +/- 0.081) U, telomerase value from 22 acute leukemia patients was (0.512 +/- 0.294) U. Telomerase activity is higher expressed in acute leukemia than normal samples and decreased significantly after chemotherapy (P < 0.01). The results suggested that telomerase activity was related to some malignant diseases and might be used as a marker for tumor diagnosis and therapy. PMID- 12578586 TI - [Photochemical Effect of the Photosensitizer Hemoporphyrin Derivative on MCF-7 Cell Line and Umbilical Cord Blood Hematopoietic Cells] AB - To study the effect of hemoporphyrin derivative(HPD) combined with laser irradiation on human breast tumor cell line MCF-7 and normal human umbilical cord blood-derived hematopoietic cells by using MTT assay. The results showed that HPD plus laser irradiation was more efficient for killing MCF-7 cells than normal human umbilical cord blood-derived hematopoietic cells. The photochemical effect with laser irradiation were higher than with light irradiation and it's effect on MCF-7 cells was higher gradually with the increase of HPD concentration. PMID- 12578587 TI - [Research on Ex Vivo Hematopoiesis Supported by Microcarriers] AB - To explore methods of maintaining the self-renewal capacity of hematopoietic stem cells, inhibiting their overdue differentiation and expanding hematopoietic cells massively, the murine bone marrow stromal cells were coated on microcarriers, then co-cultured with hematopoietic cells from murine bone marrow as group 2 (G2). The G2 contents were wrapped by sodium alginate, then cultivated as group 1 (G1). The only microcarriers coated with stromal cells as group 3 (G3) and the only bone marrow cells as group 4 (G4) were cultivated as control groups. Contrasting observation and microphotograph were performed; the number of total marrow cells, the colony efficiency of CFU-GM and the percentages of CD34(+) cells were determined. Three repeated experiments indicated that the colony efficiency of CFU-GM before culture (G0) were 118.8 +/- 38.1/10(5) marrow cells, and the total outputs of CFU-GM (G0) were 9 501.3 +/- 3 049.0. After culture for two weeks, hematopoietic cells were adhered to or embedded in stromal cells coating the microcarriers, and had formed hematopoietic focus. The colony efficiency of CFU-GM per 10(5) mononuclear cells in group G1, G2, G3 and G4 averaged 30.9 +/- 13.7, 147.3 +/- 66.0, 23.4 +/- 23.1 and 15.9 +/- 8.1, respectively; the total outputs of CFU-GM in group G1, G2, G3 and G4 averaged 273.8 +/- 75.3, 9 424.8 +/- 7 933.7, 419.1 +/- 305.6 and 140.7 +/- 20.7, respectively; the measured CFU-GM output in group G2 was significantly higher than that in group G4, and still significantly higher than the sum of groups G3 and G4 (t = 6.553, t = 5.494; P < 0.05). The percentage of CD34 cells before culture was 10.0 +/- 1.0; after cultuer for two weeks, the percentages of CD34(+) cells in G1, G2, G3 and G4 averaged 4.0 +/- 1.0, 11.0 +/- 1.0, 3.3 +/- 1.5 and 2.2 +/- 0.8, respectively. The percentage of CD34 positive control (3T3 cells) was 17.0 +/- 1.0. This result was consistent with the result of CFU-GM outputs measured. These data suggest that microcarriers coated with stromal cells can perfectly support the ex vivo hematopoiesis at least to four weeks, while hematopoietic cells fixed by alginate are not significantly different from control groups. The hematopoiesis-simulating model of microcarriers is successful, whereas the hematopoiesis-simulating model of alginate macrocarriers can not support the ex vivo hematopoiesis. PMID- 12578588 TI - [Morphology and Immunophenotype Study of Bone Marrow Mononuclear Cells after Activated by Different Cytokines In Vitro] AB - Bone marrow cells in cultures were divided into four groups and cultured with various cytokines in vitro. These four groups are: control, IL-2 group, CD3-AK group, and CIK group. The morphological (cell volume, nucleus/plasm) changes of bone marrow cells in culture were observed. Immunophenotype analysis (CD34, CD38, CD3, CD56) were done before and after culture in all groups. Cytotoxicity against fresh acute leukemia cells were detected by modified MTT methods. The cell volume became larger with increased nucleus/plasm ratio in IL-2 group, CD3-AK group and CIK group. The plasm filled with PAS positive granules in most of cells in CD3-AK group and CIK group. The positive ratio of CD3, CD56, CD38 in CD3-AK or CIK group increased markedly after culture (P < 0.05), but no significant difference between the two groups. The CD56(+) cell increased in IL-2 group. CD34(+) cells decreased in all groups and there were no significant differences among those four groups. The cytotoxicity to fresh leukemia cells: CD3-AK group and CIK group > IL-2 group > control group. There was no significant difference between CD3-AK group and CIK group. This experiment showed different effect on bone marrow cells by different cytokine combination. The cytokine combination of CD3-AK group or CIK group can make immunocytes of bone marrow proliferating and retained certain amount of stem/progenitor cells. PMID- 12578589 TI - [Effect of Autocrine VEGF on Chronic Myeloid Leukemia Cell Line K562] AB - Vascular endothelial growth factor (VEGF) is a specific mitogen for vascular endothelial cells. It has been associated with angiogenesis, growth, metastasis and poor prognosis in solid tumors. Lately, it has been known that VEGF expression is higher in bone marrow from chronic myeloid leukemia (CML) patients than that in normal subjects. However, it is not clarified that the effect of VEGF on the abnormal proliferation of CML cells. In order to explore the effect of autocrine VEGF on CML cells, K562 cells were transfected with the VEGF(121) cDNA sense vector (K562/S) or with the VEGF(121) cDNA antisense vector (K562/As). K562 cells were transfected with the pcDNA(3) vector (K562/V) as the control. Cell proliferation was determined by MTT and colony forming assay in vitro. Flow cytometric Annexin-V-FITC/PI dual labeling technique was performed to observe the effect of VEGF(121) cDNA transfection on apoptosis of K562 cells. Results indicated that K562/S transfectants exhibited a 3-fold increase in VEGF secretion, and K562/As transfectants exhibited a 49% reduction in VEGF secretion. K562/As showed a reduced growth rate and colony forming efficiency as compared to K562/V. K562/S showed an increasing growth rate and colony forming efficiency as compared to K562/V. K562/As had more apoptotic cells than K562/V and K562/S in the same culture condition. These data suggest that VEGF plays an important role in the abnormal proliferation and apoptosis in CML cells through an autocrine mechanism. PMID- 12578590 TI - [Low Concentrations of STI571 Enhances beta1 Integrin Mediated Inhibitory Effect on Proliferation of Myeloid Progenitors in Ph(+)Chronic Myeloid Leukemia] AB - To investigate whether ABL specific tyrosine kinase specific inhibitor STI571 can restore beta1 integrin mediated negative effect on Ph(+) chronic myeloid leukemia(CML), the inhibitory effect of beta 1 integrin activator (beta1 integrin activating antibody 8A2, cytokines such as GM-CSF, G-CSF and SCF) and/or FN on the granulocyte-macrophage colony forming unit (CFU-GM) from 16 patients with Ph(+)CML and 13 normal individuals were examined; the bone marrow mononuclear cells (BMMNC) before and after ABL kinase specific inhibitor STI571 pretreatment (0.1 micro mol/L for 30-60 minutes) were target cells in this study. The roles which VLA4 and VLA5 played in this process were evaluated through blocking assay. The results showed: (1) beta1 integrin activator(s) or FN alone have no effect on CFU-GM from CML or normal bone marrow mononuclear cells before or after STI571 pretreatment, nor STI571 pretreatment itself. (2) The inhibitory effect of beta1 integrin activator(s) plus FN on CML CFU-GM are significantly lower than that on normal CFU-GM. (3) The inhibitory effect of beta1 integrin activator(s) plus FN on CML CFU-GM after STI571 pretreatment is comparable to that on normal CFU-GM. (4) Monoclonal antibody to VLA4 and VLA5 or to total beta1 integrins almost completely abrogate the above effect of STI571. The results suggested enhancing beta1 integrin mediated negative effect on myeloid progenitors in Ph(+)CML is one of the therapeutic mechanisms of STI571 on Ph(+)CML. PMID- 12578591 TI - [Purification of Human CD40-Ig Fusion Protein and Its Effect on Human Peripheral Blood CFU-T Formation In Virto] AB - CD40/CD40L interactions play a pivotal role in T cell activation, and take part in many physiologic and pathologic procedures and different levels. In this article, stable CHO transformants secreting human CD40-Ig fusion protein were established through transfection and selection with Lipofectamaine and G418, respectively. In order to obtain great valume of recombinant protein, big batch serum-free cultures of engineered CHO cells were performed in roller-bottle using CHO-II-SFM medium. After cultures, the cell-culture supernatants were harvested, concentrated through ultra-filtration, and finally purified by affinity choromatography with Protein G Sepharose Fast Flow. Human peripheral bloods were collected freshly and seperated with Ficoll, CFU-T was cultured in semi-solid culture system with peripheral blood mononuclear cells (PBMNC). Effect of human CD40-Ig fusion protein on the formation of CFU-T was observed in vitro. The results showed that the yield of human CD40-Ig fusion protein was 30 mg in total 3 liter CHO-II-SFM culture supernatant, and it supposed that the expression level of CD40-Ig in CHO cells was more than 10 micro g/ml. The purity of purified fusion protein is above 95%. Furthermore, compared with human IgG, human CD40-Ig fusion protein significantly inhibited the formation of CFU-T at dose 0.25, 1.0, 4.0, and 10 micro g/ml, it lays a good foundation to evaluate its potential functions in vivo. PMID- 12578592 TI - [Effect of Inhibition of Telomerase on Cisplatin-Induced Apoptosis in HL-60 Cells] AB - To explore the effect of inhibition of telomerase activity on cisplatin-induced apoptosis in HL-60 cells, polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) was used to determine telomerase activity in HL-60 cells untreated or treated with human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN); apoptosis was detected by morphological observation and cell cycle analysis by flow cytomertry. Telomerase activity in HL 60 cells treated with hTERT ASODN was shown to decrease with time of treatment. The combination of hTERT ASODN and cisplatin could induce apoptosis of HL-60 cells. Morphological characteristics of apoptosis, such as cell shrinkage, chromatin condensation, nuclear segmentation and formation of apoptotic bodies, were observed. The percentage of apoptotic HL-60 cells treated with cisplatin for 48 or 72 hours after 24 hours of exposure to ASODN was significantly increased than that of groups with sense oligodeoxynucleotide (SODN) plus cisplatin or cisplatin alone (P < 0.01), respectively. However, there was no difference in percentage of apoptotic HL-60 cells between hTERT ASODN plus DNR or Ara-c, SODN plus DNR or Ara-c, and DNR or Ara-c alone, respectively. The results here demonstrated that inhibition of telomerase activity by hTERT ASODN could enhance the cisplatin-induced apoptosis of HL-60 cells. PMID- 12578593 TI - [Alteration of the Cell Cycle during the Differentiation of HL-60 Cells by Induction of Retinoic Acid] AB - To study the alteration of the cell cycle during the differentiation of human myeloid leukemia cell line HL-60 induced with all trans-retinoic acid (RA), the flow cytometry was used to assay the various phases of cell cycle in HL-60 cells treated with RA. The results showed: (1) S + G(2)/M phase proportion kept relative invariability during the 48 hours incubation of HL-60 cells with RA, however, the proportion alteration of S-phase cells was associated with the RA concentrations. At 10(-6) mol/L RA, the proportion of S-phase cells appeared a temporarily increasing peak followed by persistent decrease of S-phase proportion. At 10(-5) mol/L RA, S-phase cell proportion only appeared the persistent decreasing tendency. (2) Re-culture of HL-60 cells without RA showed the decrease of S + G(2)/M and S-phase cells was associated with the increase of differentiated cells, but not all HL-60 cells were triggered into differentiation at the same time. Once the cells start to differentiate, even if there is no RA presence, the HL-60 cells were still differentiated until maturation. In conclusion, HL-60 cells are able to differentate and maturate after exposure to RA for a period of time. S-phase proportion is related to the concentrations of RA. Once the cells start to differentiate, even if there is no RA presence, the HL-60 cells still differentiatiated until maturation. PMID- 12578594 TI - [Effects of Red Orpiment on Cell Morphology and Expression of PML mRNA and Protein in NB4, and HL-60 Cells] AB - In this study the effects of red orpiment on NB4 and HL-60 cells were tried to determine. Semi-quantitative RT-PCR to determine the PML mRNA expression, immuno fluorcscence study, together with the fluorescence stain and morphological observations were used. The results showed that: (1) red orpiment induces apoptosis morphologically in NB4 and HL-60 cells, the morphology of typical apoptosis can be seen in NB4 and HL-60 cells after 12 hours of treatment with red orpiment. Through the Wright's stain, we can see the extensive membrane blebbing, condensation of heterochromatin, and cell fragmentation, apoptotic body appearing. Many dead cells can be found on the second day. (2) in NB4 cells, red orpiment is shown to induce the PML-RARalpha chimera disappearance and to reorganize then to degradation of PML nuclear bodies which also seen in HL-60 cells, indirect immunofluorescence staining of PML with a specific monoclonal antibody was performed in control and treated cells. In NB4 cells, the control was diffusely microspeckled pattern of immunoreactivity. Upon red orpiment treatment, the microspeckled pattern disappeared, PML protein reversed into normal location. and the the size and the brightness of the particles were increased obviously. The normal nuclear distribution of PML protein was seen in untreated HL-60 cells. After treatment with red orpiment, in the nuclei of HL-60 cells, the size and the brightness of the particles were also increased. After two days of treatment with red orpiment, the immunofluorescent particles in cells almost disappeared. (3) the expression of PML mRNA is not changed in red orpiment treated cells, RT-PCR to determine the PML mRNA expression in NB4 and HL-60 cells treated with red orpiment, the expression results are similar to the controls, that to say, the PML mRNA lever is unaffected. It was concluded that, red orpiment induced PML to play the effects of induce apoptosis in leukemia cells at the translational level and inhibited the proliferation of leukemia cells. PMID- 12578595 TI - [Clinical Study on A Patient with Intestinal Acute Graft-Versus-Host Disease Grade IV after Allo-Peripheral Blood Stem Cell Transplantation] AB - The purpose of this study was to designed to analyze and explore the diagnosis and treatment on a patient with beta-thalassemia major accompanied with intestinal acute graft-versus-host disease (aGVHD) grade IV after allo-peripheral blood stem cell transplantation (allo-PBSCT). The conditioning regimen was consisted of busulfan (18 mg/kg), cyclophosphamide (200 mg/kg), thiotepa (6 mg/kg) and horse anti-thymocyte globulin (110 mg/kg). The nucleated cells (10.6 x 10(9)/kg) and CD34(+) CD38(-) cells (11.7 x 10(6)/kg) were infused. At +14 days PBSCT, ANC and WBC achieved 0.66 x 10(9)/L and 1.4 x 10(9)/L respectively. The engraftment was successful but the patient depended on transfusion. The range of blood WBC was from 1.4 x 10(9)/L to 2.4 x 10(9)/L. The myelogram showed aplastic anemia. Hematopoiesis was restored continually after the second transfusion of the donor's cells. At +11 days the patient was suffered from cutaneous aGVHD grade II. Diarrhea occurred at +33 day when cutaneous aGVHD had been improved, and exaggerated obviously after the second transfusion at +39 days. The serious diarrhea continued for 2 months and intestinal aGVHD grade IV was finally diagnosed. Our conclusion was that intestinal aGVHD possibly occurred even when cutaneous aGVHD was improved. For a patient with refractory diarrhea after allo PBSCT, intestinal aGVHD was considered first when the engraftment evidence was definited but peripheral blood cells and bone marrow were abnormal. PMID- 12578596 TI - [The Effects of Cyclosporin A on Leukemia Cells In Vitro] AB - To explore the mechanism of CSA in the reversal of drug-resistance of leukemia cells, K562 and K562/ADM cells were used for in vitro study. Drug sensitivity of cells was evaluated by MTT assay and cell survival by trypan blue exclusion. Apoptosis was detected by terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL) and agarose gel electrophoresis. The results showed that 1.0 mg/L CSA alone have no effect on the survival of K562 and K562/ADM cell lines, but it could enhance the chemotherapy sensitivity of the 2 cell lines, and accelerate the apoptosis induced by ADM. These effects were stronger in K562/ADM cell line, when the concentration of CSA > 1.0 mg/L, CSA itself could kill leukemia cells and induce the apoptosis of the 2 cell lines. It is concluded that CSA can reverse leukemia cells' drug-resistance by enhance apoptosis of the leukemic cells, CSA itself can induce leukemia cells' apoptosis and kill leukemia cell too. PMID- 12578597 TI - [GPI-Pr Deficiency and Apoptosis of PNH Granulocytes] AB - To study the relationship of Glycosyl phosphatidylinositol anchored proteins (GIP Pr) and apoptosis of paroxysmal nocturnal hemoglobinuria (PNH) cells, we isolated peripheral granulocytes from 10 patients with PNH and 10 normal controls and measured apoptosis induced by serum starvation. The FCM analysis of phosphotidylserine (ps) externalization in granulocytes was determined using Annexin-V-FLUOS labeling. After the cells were induced for apoptosis in serum free medium for 20 hours, the percentage of externalization was 78.6% in normal control cells but 39.5% in PNH cells. The results of FCM analysis of PI stained granulocytes showed that the PI positive rate was 51.5% in control cells and 30.2% in PNH cells. The gel electrophoresis analysis of DNA fragmentation all indicate that PNH granulocytes were relatively resistant to apoptosis as compared with normal controls. This resistance to apoptosis might not be related to the percentage of CD59 deficient granulocytes. PMID- 12578598 TI - [The Effect of Low Molecular Protein Iron on Iron Deficiency Anemia] AB - Objective is designed to observe the effect of the low molecular protein "SL Hong Xin Blood-Increasing Capsules" on human iron deficiency anemia. Altogether 110 school children, 7 - 12 years old, with iron deficiency anemia (Hb < 120 g/L) were enrolled in this trial and were randomly and evenly divided into two groups. In the treatment group (n = 55), the "SL Hong-Xin Blood-Increasing Capsules" were orally adminitered for 30 days, two capsules at one time and two times a day, whereas in the control group (n = 55) blank capsules were given to the children in the same way as in the treatment group. Before and at the end of the trial period, peripheral blood samples were taken from fringers of the children to determine Hb content, hematocrit percentage, and free protoporphyrin content in red blood cells. Results showed that after trial the Hb content was (15.9 +/- 9.6) g/L in the treatment group, while that of the control group was only (5.3 +/ 4.3) g/L (P < 0.001). The hematocrit percentage markedly increased (P < 0.001) and protoporphyrin level markedly decreased (P < 0.001). In conclusion the active ingredient of "SL Hong-Xin Blood-Increasing Capsules" is low molecular protein iron, which is markedly effective for elevating Hb content and hematocrit percentage, and effective for decresing protoporphyrin content of children with iron deficiency anemia. Hence, the capsules could be used to improve nutritional anemia in children based on the "Functional Evaluation Procedure and Test Methods For Health-Care Food". PMID- 12578599 TI - [Donor Hematopoietic Cell Tracking In Vivo at the Homing Phase of Allo-Bone Marrow Transplantation in Mice] AB - It has been well-known that intravenously infused hematopoietic stem and progenitor cells can home to the bone marrow and reconstitute hematopoiesis. However, little is understood about the homing efficiency or percentage of infused stem and progenitor cells. In order to examine distribution pattern of infused hematopoietic cells in the organs and tissues, a direct assay system to trace transplanted cells in vivo by employing PKH-26, a red fluorescent membrane dye, to label hematopoietic cells in inbred strain of mice transplanted cells (stem cell antigen-1 positive subpopulation cell, Sca-1(+) cells) was introduced. The numbers of labeled cells was measured by means of flow cytometry and fluorescence microscopy. The early fate of infused Sca-1(+) donor bone marrow cells after intravenous administration in a allogeneic mouse model was examined. The presence of infused donor cells with the fluorescent dye PKH-26 was evaluated within 60 hours in hematopoietic organ (bone marrow and spleen) and non hematopoietic organ (lungs and liver) of recipients. The data showed that (1) Following intravenous infusion, Sca-1(+) donor bone marrow cells were detained in lungs shortly. (2) Sca-1(+) donor bone marrow cells localized to both hematopoietic organ (bone marrow and spleen) and non-hematopoietic organ (lungs and liver) for periods of up to 60 hours following infusion, however, the number of donor hematopoietic cells localized to bone marrow was more than that localized to non-hematopoietic organ (P < 0.05). These results indicated that there were also donor early hematopoietic cells in non-hematopoietic organ of recipients at the homing phase in allo-BMT mice. PMID- 12578600 TI - Plasma Soluble Fas Ligand Levels in Hemopathic Patients Undergoing Allogeneic Bone Marrow Transplantation. AB - To evaluate the implications of soluble Fas ligand (sFasL) in acute graft-versus host disease (aGVHD) and discriminating symptoms of aGVHD from those of infection, plasma levels of sFasL were assessed in 84 plasma samples from 13 patients who had undergone allogeneic BMT by using a sandwich enzyme-linked immunological assay (ELISA). Plasma sFasL levels of the patients before BMT and at different time points during the post-BMT period were measured. Three points were concluded: (1) Plasma sFasL levels were higher in patients with grade II-IV aGVHD than in those with grade 0 or I aGVHD. (2) Plasma sFasL levels in patients with infection were not statistically different from those in patients without infection. (3) In seven patients with grade II-IV aGVHD, the plasma sFasL levels at pre-BMT were much lower than those of the six patients with 0 or I grade aGVHD. sFasL may be useful for the diagnosis of aGVHD and for differentiating aGVHD from other BMT related complications such as infection. PMID- 12578602 TI - Analysis of Human Platelet Antigen Genotypic Frequencies in Chinese Population by PCR Amplification with Sequence Specific Primers. AB - In present study, a method for genotyping for human platelet antigen (HPA) systems by means of the polymerase chain reaction amplification with sequence specific primers (PCR-SSP) technique was developed and employed to determine the human platelet antigen frequencies in the Chinese population. Primers sets were designed to allow PCR amplification for five systems using the same assay conditions. Platelets from 110 random Chinese blood donors were typed for human platelet alloantigens HPA-1 to -5 by the method. The results showed that the HPA genotypic frequencies observed in the 110 donors were 0.91 and 0.09 for HPA-1a and HPA-1b, 0.86 and 0.14 for HPA-2a and HPA-2b, 0.60 and 0.40 for HPA-3a and HPA 3b, 0.92 and 0.08 for HPA-4a and HPA-4b, and 0.85 and 0.15 for HPA-5a and HPA-5b, respectively. In conclusion the method is feasible and practical and may be available to typing for HPA in the clinical laboratories. PMID- 12578601 TI - [Research of Typing for HLA-A, -B on Cord Blood Lymphocytes] AB - Serological typing for HLA-A, -B has been used for a long time. Recently with the developing of molecular biology technologies, HLA-A, -B typing is now turning to genotyping methods. In our study, the capacity of PCR-SSP in solving problems in HLA-A, -B typing with serological methes was evaluated. With this aim the serological method was compared with PCR-SSP in 102 cord blood samples, and the results showed that 18.6% of 102 cord blood samples can't give a satisfactory detection, for 14 samples, give discrepant results with the 2 methods. It is mainly due to weak expression of HLA class I cord blood lymphocytes and the cross reaction of some antigens. About B 15 group, the further study was made, it was found that most of the B 15 splits is wrongly disassigned, especially among the B62-B75, B75/*1511(+)-B75/*1511(-), B46-*1511 antigens. It was concluded that DNA typing is more preferable than serological typing, about B 15 group, the subtyping or high resolution typing can be fulfilled at first in China. PMID- 12578603 TI - [The Examination Method of Glycoprotein CD62P and CD63 on Platelet Membrane and Their Expression in Adults Patients with Diabetes Mellitus] AB - To study the optimal examination method of CD62P and CD63 and investigate platelet activation in patients with diabetes mellitus (DM), whole blood labeled directly with monoclonal antibodies CD62P and CD63 and flow cytometry were used to evaluate the positive percentages and the mean fluorescence intensity of CD62P and CD63. The specimens of peripheral blood obtained from 10 healthy adults were divided into two groups. In the unfixing group, the positive percentages of CD62P and CD63 at the periods of 30, 60, 90 and 120 minutes after staining were (7.57 +/- 2.33)%, (20.50 +/- 5.70)%, (28.70 +/- 5.67)% and (36.52 +/- 6.13)%, and, (0.89 +/- 0.36)%, (1.11 +/- 0.84)%, (2.35 +/- 2.02)% and (5.43 +/- 3.66)% respectively, their respective MFI were 1.57 +/- 0.13, 1.88 +/- 0.08, 2.00 +/- 0.09 and 2.38 +/- 0.22 and 3.91 +/- 0.11, 4.07 +/- 0.16, 4.38 +/- 0.14 and 4.44 +/- 0.19. However, in fixing group with 1% paraformaldehyde, the results had not any obvious change and almost were same. Besides it, the positive percentages of CD62P and CD63 in 37 adult patients with DM were (14.11 +/- 6.68)% and (2.71 +/- 1.74)%, significantly higher than that in the normal controls. It is concluded that the CD62P and CD63 on platelet membrane were very sensitive and would be easily activated in vitro, all manipulations that includes labeling with antibody, incubation and detection using flow cytometry should be finished within 30 minutes after samples collected. While fixing by using 1% paraformaldehyde can steady the labeling compounds and effectively prevent the artificial activation of platelet, and keep the stable results within two hours after the samples labeled. In adult patients with DM, the relationship between the cardiovascular complication of diabetes and platelet activation might be existed. PMID- 12578604 TI - [Study on Polyethylene Glycol Modified Porcine Hemoglobin Using Computer Modeling] AB - The structure analysis of porcine hemoglobin alphabeta dimer and the calculation of solvent accessible surface of the amino acids showed the epsilon-amino groups of the lysine are suitable for modification by polyethylene glycol (PEG). The modification of the lysine residues will not affect the carring oxygen capacity of Hb. Three types of linker have been designed to connect PEG and porcine hemoglobin. The lysines between porcine and bovine hemoglobin (pHb and bHb) are highly conserved, but the solvent accessible surface of conserved lysines are different. These suggested that the properties of homologous proteins are similar in pHb and bHb, but the characteristic derived from the homology analysis will be deviated from the actual status. The results of molecular dynamics simulation suggested that the chemical modified porcine hemoglobin would be no immunogenicity. PMID- 12578605 TI - Vascular Endothelial Growth Factor and Its Receptor KDR/flk-1 Play Important Roles in Hematopoiesis. AB - It is widely accepted that hematopoietic and endothelial cell lineages are initiated from the same precursor cells (named as hemangioblast), although hemangioblast has not been proved. Vascular endothelial growth factor (VEGF) and its receptor KDR/flk-1 is a prime regulator of endothelial cell proliferation, angiogenesis, vasculogenesis and vascular permeability. It is speculated that VEGF and its receptor KDR/flk-1 may also play an important role in embryonic and postnatal hematopoiesis. But the exact role and mechanism are not well known. Some latest developed cellular and molecular techniques can be used to prove the existence of hemangioblast and to investigate its biologic feature and the effect of VEGF and receptor KDR/flk-1 on its biologic feature. PMID- 12578606 TI - [p73 Gene and Leukemia] AB - p73 has high homology with the classical tumor suppressor gene p53. When overexpressed, p73 could lead to tumor cells growth arrest and induce apoptosis. Lost of p73 gene expression due to hypermethylation was found in about one third patients with acute lymphoblastic leukemia and Burkitt's lymphoma, but the expression level of p73 was higher in patients with acute myelogenous leukemia than that in normal white blood cells. Further research will be necessary to find out the role of p73 in leukemogensis. PMID- 12578607 TI - [Real-Time Quantitative PCR and Detection of Minimal Residual Disease in Blood Malignancy] AB - Real-time quantitative PCR (RQ-PCR) is a recently developed technique for nuclear acid quantification, in which conventional PCR and probe hybridization are integrated together. A specific fluorescent-labeled probe, binded with DNA/cDNA target, is supplemented into the reaction system of PCR. During the extension phase of PCR amplification, the specific fluorescent-labeled probe is cleaved by the 5'-->3' exonuclease activity of Taq polymerase, producing a fluorescent signal that can be detected. The amount of fluorescence released in the reaction is proportional to the target sequence number. Thus, an exactly and absolutely quantitative assessment of initial target DNA/cDNA copy number can be made in a closed tube assay system. This technique eliminates the need for post-PCR sample processing which is necessary in conventional PCR and decreases the probability of contamination which is inevitable in conventional PCR. Moreover, it reduces the potiental for false positive or negative results by utilizing the hybridization of a probe to sequences internal to amplification primers. RQ-PCR is proved to be a sensitive, specific, reliable and convenient tool for the quantification of minimal residual diseases (MRD) of hematological malignancies by recent studies. This article reviews the principle of RQ-PCR and its application in quantitative detection of MRD of hematological malignancies such as acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML) and malignant lymphoma. PMID- 12578609 TI - [The Effect of Different Doses of Aspirin on Platelet Function and Its Clinical Importance] AB - The aim of this study was to investigate the initial time of platelet inhibiting effect of aspirin (ASA) and the effects of different doses on equilibrium of prostacyclin (PGI(2))-thromboxane B(2) (TXB(2)). The effects of 100 mg and 300 mg ASA on Platelet count, platelet aggregation rate, TXB(2) and PGI(2) were investigated using cross-compare way for 40 aspirin ingestion patients. The results showed that the platelet counts decreased to 33% after 30 minutes of single-dose ASA ingestion of 100 mg and to 25.6% after 60 minutes. TXB(2) and PGI(2) also decreased meanwhile. The platelet counts decreased to 39.5%, 35.5% and 26.6%, respectively with dose of 300 mg on day 1, 2 and 3. The platelet counts decreased to 38.1% and 39.5%, respectively, after 120 minutes with 100 and 300 mg ASA ingestion, without significent difference. In conclusion ASA began to inhibit platelet function after 30 minutes of ingestion, and gave the strongest inhibition after 60 minutes. Continuous ASA ingestion accumulates the inhibitory effect. The single-dose ASA ingestion of 100 and 300 mg have nearly the same inhibitory effects. PMID- 12578608 TI - [Nonmyeloablative Allogeneic Peripheral Blood Hematopoietic Stem Cell Transplantation for Therapy of Hematologic Diseases] AB - The objectve of this investigation was to explore the use of nonmyeloablative allogeneic peripheral blood stem cell tranplantation for treatment of hematologic diseases. Six patients were included: 3 cases with acute leukemia in first complete remission (2 AML and 1 ALL), 2 severe aplastic anemias and 1 chronic myeloid leukemia in chronic phase. All of the 6 cases were received HLA-identical siblings donor peripheral blood stem cell transplantation after a nonmyeloablative conditioning. The donor cells were engrafted in all patients (3 cases were full engrafment of donor cells and 3 were mixed chimerism). Hematopoietic recovery was appeared in all of the cases (ANC recovered to more than 0.5 x 10(9)/L and platelet count to more than 30 x 10(9)/L on day 9 to day 21 and day 14 to day 28 after transplantation, respectively). Two patients developed III or IV degree GVHD. Our preliminary results suggest that the procedure is more safe, efficient and less complications than myeloablative conditioning regimens and represents another new approach in the management of patients with hematologic diseases. PMID- 12578610 TI - [GVHD Following Autologous Peripheral Blood Stem Cell Trasplantation Reduced Malignancy Relapse] AB - Administration of the immunosuppressive drug cyclosporine (CSA) after autologous peripheral blood stem cell transplantation (APBSCT) induces a systemic auto immune syndrome resembling graft-versus-host disease (GVHD), this syndrome termed autologous GVHD has significant antitumor activity, it can reduce the incidence of tumor relapse after APBSCT. The antitumor effect of this auto-aggression syndrome can be enhanced by the administration of gamma-interferon (gamma-IFN). Five consecutive patients who received APBSCT received therapy inducing autologous GVHD. Intravenous administration of CSA [1 mg/(kg.d) for 28 days] was begin on the day of transplantation. gamma-interferon (0.025 mg/m(2) qod) was administered sub-cutaneously from days 7 throngh 28 after transplatation. Results showed that four of five occured autologous GVHD-skin demage, five in control didn't occur autologous GVHD. The relapse rate of the treated cases was 20% (1/5) versus 60% (3/5) of the control, and the median survival time of the treated cases was 20 (4 - 30) months versus 10 (2 - 20) months of the control. The data indicates that autologous GVHD results in low relapse rate of the patients rececving APBSCT. PMID- 12578611 TI - [Maintaining Growth of Long-Term Culture Initiating Cells from Human Cord Blood on Feeder Layers of Bone Marrow Stromal Cells Transfected with FL and/or TPO Genes] AB - Long-tem culture initiating cells(LTC-IC), and in vitro assay of hematopoietic stem/progenitor cells, still represent a heterogeneous population in terms of proliferative capacity and sensitivity to different growth factors. Human umbilical cord (CB) is rich of hematopoietic progenitor cells measured by clonogenic assays and stem cells capable of reconstituting the marrow after transplantation. The influence of culture conditions on the in vitro behavior of LTC-IC from CB was evaluated. First, by using IRES sequence, FL and TPO cDNA were recombined with retroviral vector pLXSN by gene recombination technology. The recombinant plasmid pLFSN, pLTSN, pLFTSN were transfected into human stromal cell line HFCL. Then, LTC-IC were evaluated in long term cultures, comparing five types of stromal feeder layers: human bone marrow stromal cell, human stromal cell line HFCL, and stromal cell lines HDF tranfected with FL gene, HLT transfected with TPO gene or HFT co-transfected with FL and TPO genes. The results were demonstrated that after 8 weeks of coculture, three types of stromal cell lines that supported the maintenance of CFU-C for up to 3 weeks in vitro were identified. However, cocultivation of human bone marrow stromal cell and CB CD34(+) cells on HFT, CFU-C production continued up to 6 weeks or longer on these stroma. The absolute LTC-IC frequency in CD34(+) cells on human bone marrow stromal cell (2.65 +/- 0.76/1 000 cells) was no significant difference with on HFT (3.65 +/- 0.58/1 000 cells). Thus, HFT acts by direct contact to maintain the phenotype and function of the most primitive and quiescent human progenitors. Furthermore, HFT cell line was selected as the optimal one for supporting long term culture feeder. It was concluded that LTC-IC progenitors from cord blood maintain growing upon the FL/TPO gene-modified stromal feeder layers in vitro. PMID- 12578612 TI - The Presence of Endothelial Cell Precursors in Blood Circulation. AB - In the present study, an attempt was made to prove the question whether endothelial cell precursors exist in blood circulation during postnatal period. CD34(+) cells were harvested from G-CSF mobilized adult blood and umbilical cord blood and incubated onto fibronectin/gelatin-coated Petric dishes in the presence of recombinant human vascular endothelial cell growth factor(rhVEGF) and basic fibroblast growth factor(rhbFGF). Endothelial cell lineage was identified by von Willebrand factor(vWF) expression and Ulex europous agglutinin I(UEA-I) binding capacity. The results showed that a firmly adherent cell monolayer formed when CD34(+) cells, but not CD34(-) cells, were cultured for 5 - 6 weeks as described before. Immunocytochemistry and flow cytometry analysis showed that almost all of the adherent cells were vWF-positive and around 90% were able to bind UEA-I specifically. These findings demonstrate that angioblasts exist in the circulation during postnatal life and therefore, vasculogenesis might occur in adults. PMID- 12578613 TI - [Induction of Apoptosis in Leukemic Cells by Inhibiting the Ubiquitin-Proteasome Pathway and Its Possible Mechanism] AB - The ubiquitin-proteasome pathway is the principal mechanism for the degradation of short-lived proteins in eukaryotic cells. Recently, proteasome inhibitors have been shown to induce apoptosis in many kinds of human malignant cells. In this study, the mechanism of apoptosis induced by proteasome inhibitor in leukemic cells was examined. Evaluated by MTT assay, treatment of leukemic cells with Z LLL-CHO, a reversible proteasome inhibitor, induced cell death in a dose dependent manner. Appearance of the sub G(0)/G(1) fraction of cell cycle observed in flow cytometry assay suggested the induction of apoptosis, which was further proved by typical DNA ladder and morphological study. Western blot displayed the cleavage of bcl-2 into a shortened 22 kD fragment and the decrease in the levels of caspase-3 precursor. A highly sensitive colorimetric assay was employed and the elevation of caspase-3 activity was detected in both cell lines after treatment with Z-LLL-CHO. By comparison, these results showed that the leukemic cell line M-07e and KG-1a, which both express bcl-2 at a relative high level, had different susceptibility to undergo apoptosis induced by Z-LLL-CHO, which possibly due to their different levels of expression and activation of caspase-3 precursor, as well as their different degree of bcl-2 cleavage after treated by Z LLL-CHO. PMID- 12578614 TI - [Erythropoietin Receptor-Mediated Proliferative Signal Transduction in Leukemic Cell Line KOCL-33] AB - In order to investigate the expression of erythropoietin receptor(EPOR) in leukemic cell line and clarify the mechanism of proliferative signal transduction mediated by EPOR in the leukemic cell line KOCL-33, biotinylated EPO and flow cytometry were used for detection of EPOR expression, (3)H-TdR incoporation for cell proliferation, immunoprecipitation and Western blot analysis were used to investigate the tyrosine phosphorylation of signal transduction proteins as the causation of enhanced proliferation of KOCL-33 cells. There was EPOR expression and the cell proliferation could be stimulated by EPO in KOCL-33 cells. Dimerization and tyrosine phosphorylation of EPOR were found after 1 minute incubated with EPO. Coimmunoprecipitation of SHC and PLC-gamma and enhanced tyrosine phosphorylation following to the stimulation of EPO were found. Our results suggest that dimerization of EPOR and tyrosine phosphorylation were appeared under the stimulation of EPO, and SHC and PLC-gamma play a cooperative effect in EPOR mediated signal tranduction. PMID- 12578615 TI - [p21(WAF1) Gene Transfection Inhibits Proliferation of Leukemia Cell Line K562] AB - To explore the functional role of p21(WAF1) gene on the proliferation of leukemia cell line K562, a p21(WAF1) retroviral expression vector was constructed. Mediated by FuGENE trade mark 6, p21(WAF1) was transfected into leukemia cell line K562, which was without p21(WAF1) expression. After selected in G418, K562 cell clones that expressed p21(WAF1) stabaly were isolated and named K562 p21(WAF1). The ectopic expression of p21(WAF1) mRNA and protein in K562 cells was identified by RT-PCR and Western Blot. The cell proliferaton was tested in liquid and soft agar culture after transfection. The cell cycle was tested by FCM. The expression of p21(WAF1) protein and mRNA could be detected in K562-p21(WAF1) cells. A strong inhibition of cell proliferation was observed in K562-p21(WAF1) cell clones cultured in liquid media as well as soft agar (P < 0.01). The cell number in G(0)/G(1) phase was remarkably increased. The findings showed that p21(WAF1) can inhibit the proliferation of leukemia cells, and it could be a potential target gene for leukemia gene therapy. PMID- 12578616 TI - [Effects of Exogenous Wild-Type p53 Gene on K562 Cells] AB - To investigate the effects of wt-p53 gene on proliferation and differentiation of K562 cells and to explore the feasibility of wt-p53 in leukemia gene therapy, pC53-SN(3), containing wt-p53 cDNA, and temperature-sensitive p53 mutant pN53cG(Val135) which behaved like wt-p53 at 32.5 degrees C, were introduced into p53-null K562 cells respectively by lipofectin mediated DNA transfection. In the presence of G418, K-SN(3) and K-pN53cG clones expressing P53 protein were selected. The effects of exogenous wt-p53 gene on the proliferation and differentiation of K562 cells were studied by detection of cell growth curves, leukemic colony formation, cell cycle analysis and DNA fragmentation, TdT mediated dUTP nick end labeling (TUNEL) and benzidine staining. The results showed: (1) The level of p53 mRNA in K-SN(3) cells was lower than that in K pN53cG cells by RT-PCR. (2) K-SN(3) and K-pN53cG(32.5 degrees C) cells proliferated more slowly than the control K562 cells, and their colony formation was obviously suppressed. The cells in G(0)/G(1) phase increased, and the cells in S phase decreased. These features were more obvious in K-pN53cG(32.5 degrees C). (3) K-pN53cG(32.5 degrees C) showed the feature of apoptosis and K-SN(3) showed the characteristics of erythroid lineage differentiation. It was indicated that exogenous of wt-p53 was capable of inhibiting the proliferation of K562 cells and inducing apoptosis of the cells at higher p53 level and interestingly, inducing the cells differentiation on erythroid lineage at lower p53 level. PMID- 12578617 TI - [The Antisense Oligodeoxynucleotides of bcl-2 Oncogene Enhances the Sensitivity of K562 Leukemic Cells to As(2)O(3)] AB - The biological reaction, changes of cellular DNA and bcl-2 protein content of K562 cells to bcl-2 ASODN or As(2)O(3) alone or bcl-2 ASODN associated with As(2)O(3) were investigated by cellular microculture, immunological tissue chemistry method, and flow cytometry. The results showed that there is much more induce-apoptotic effect of As(2)O(3) on K562 cells while it combined with bcl-2 ASODN (ASODN 10.0 micro mol/L + As(2)O(3) 5.0 micro mol/L) than bcl-2 ASODN or As(2)O(3) alone (ASODN 10.0 micro mol/L or As(2)O(3) 5.0 micro mol/L), (P < 0.01), and so does inhibitory effect of bcl-2 protein expression by flow cytometry (P < 0.01). In conclusions, the bcl-2 ASODN can enhance the drug sensetivity of K562 leukemic cells to As(2)O(3). PMID- 12578618 TI - [Change of WT1 Gene Expression during Induction of Differentiation of NB4 Cell Line] AB - In order to study the relation between WT1 gene expression and differentiation of NB4 leukemic cells, a competitive RT-PCR method was established by using recombinant DNA technique to detect the expression of WT1 gene quantitatively during differentiation of NB4 leukemic cells induced by retinoic acid. The expression of CD11b was simultaneously determined by an indirect immunofluorescence staining technique and flow cytometry. Our results showed that WT1 gene expression rapidly decreased during the differentiation of NB4 cells. The molecular number of WT1 gene in 1 micro g total RNA was 4 x 10(6), 1.56 x 10(6) and 0.4 x 10(6), respectively, prior to and at 12 or 24 hours after exposure to retinoic acid, and it was in accordance with the change of CD11b. These data suggest that the abnormally high expression of WT1 gene in leukemic cells was associated with the block of cell differentiation. The detection of WT1 expression with competitive RT-PCR combined with measure of CD antigens will contribute to study on the differentiation induction of leukemic cells. PMID- 12578619 TI - [Extraction of RNA from Stored Bone Marrow Smears for Detecting Expression of the WT1 Gene in MDS Patients] AB - RNA was extracted from stored bone marrow smears, expression of WT1 gene in the patients with MDS was examined by means of nest RT-PCR. The results showed WT1 gene was highly expressed in some cases of MDS. Expression rates were higher in the patients with RAEB and RAEB-t than those patients with RA and RAS. WT1 expression is related to the advance of MDS. The detection of WT1 gene expression by RT-PCR might be useful for assessing disease progress in the patients with MDS. PMID- 12578620 TI - [The Detection of Clonal IgH Gene Rearrangement in Bone Marrow and Peripheral Blood from B-NHL Patients and Its Clinic Significance] AB - To evaluate the significance of bone marrow (BM) and peripheral blood (PB) cells with clonal gene rearrangement of the third complementary determining region of immunoglobulin heavy chain (IgHCDR3) in the diagnosis, clinical staging, determination of treatment effects and prediction of relapse in B-NHL, clonal IgH gene rearrangement of BM from 46 and PB from 38 cases with B-NHL were tested by semi-nested polymerase chain reaction (SnPCR) and polyacrylamide gel electrophoresis before treatment, and ten of them were tested in complete remission after treatment. Results showed that this method was applicable to detecting one clonal IgHCDR3 gene rearrangement positive cell from up to 1 000 normal cells. Specificity of detection was 97%. Clonal IgHCDR3 rearrangement was shown in all 3 cases of BM and 2 of PB specimens with morphologic involvement. The clonal IgHCDR3 was detected in 65.1% of the BM and 44.4% of the PB without morphologic involvement in untreated patients with B-NHL, independent of Ann Arbor staging and systemic symptoms. In 10 cases of B-NHL with clonal IgHCDR3 rearrangement in diagnostic tissues, the molecular marker became negative in 7 patients who entered and remained in complete remission. Two cases relapsed in whom clonal IgHCDR3 rearrangements were detected in serial samples of BM or PB after autologous PBSCT. One patient in whom clonal IgHCDR3 rearrangement was detected at 10 months post-PBSCT remained in complete remission up to now. It was concluded that clonal IgHCDR3 gene rearrangements were found in BM and PB from B NHL patients without morphologic abnormality. Persistence of molecular marker positive may be associated with relapse for patients in complete remission, and the patients without clonal IgHCDR3 rearrangement will be in continuous complete remission. Little is known about a few patient who was a long-term disease-free survivor despite the presence of PCR-IgH rearrangement in the marrow. PMID- 12578621 TI - [Follow up Detection of AML/ETO Fushion Transcripts after Chemotherapy or Bone Marrow Transplantation in Leukemia Patients] AB - Expression of AML1/ETO mRNA was observed in bone marrow cells from 49 untreated leukemic patients, and continuously detected during different periods after chemotherapy (12 cases) or bone marrow transplantation (8 cases). The results showed that AML1/ETO mRNA could be expressed in cells from AML-M(2), AML-M(4) and MDS-RAEB-T patients. The positive expression changed into negative at different duration in patients who achieved complete remission either by chemotherapy (9 cases), allogeneic bone marrow transplantation (5 cases) and autologous peripheral blood stem cell transplantation (1 case), and they were sustained in complete remission status. In chemotherapeutic group, patients whose AML1/ETO expression turning from negative (2 cases) or faint positive (1 case) to positive relapsed later. Two patients treated with Allo-BMT showed continuously positive results and died of GVHD and relapse, respectively. These observations suggest that AML1/ETO chimeric mRNA could disappeared after chemotherapy or bone marrow transplantation. The patients have a great probability to relapse if the results of RT-PCR are continuously positive or change from negative to positive. Regular detection is necessary for leukemic patients. PMID- 12578622 TI - [Bone Marrow Cells Activated by Autologous Dendritic Cells Purges Bone Marrow from Patients with Chronic Myelogenous Leukemia] AB - In order to investigate the effect of autologous dendritic cells (DCs) activating bone marrow cells and purging bone marrow from chronic myelogenous leukemia (CML) patients, DCs were separated by negative selection system of human cells from bone marrow mononuclear cells (BMMNCs) of 2 CML patients in hematological remission and harvested after 3 days of culture in IMDM containing autologous plasma, rhGM-CSF and rhTNFalpha at 37 degrees C, 5% CO(2) humidified atmosphere. BMMNCs from the patients were also used to set up long-term culture (LTC) system in T-25 plastic flasks. The LTCs included three groups, i.e., control, addition of rhIL-2, and co-culture with autologous DCs. Half of non-adherent cells were collected, counted and assayed for CFU-GM weekly. Then, equivalent volume of fresh medium was replaced to maintain the culture. The culture was discontinued if the non-adherent cells count was less than 2 x 10(5). Adherent cells were collected for CFU-GM assay and flow cytometry for CD34 and P210. The colonies originating from the adherent cells were picked up under the inverted microscope. RNA was extracted, and BCR/ABL measured by nested reverse transcription polymerase chain reaction (RT-PCR). The results showed that the CFU-GM yields of non-adherent cells declined after 1 to 2 weeks co-cultured with autologous DCs, and it paralleled with group with rhIL-2. P210(+) cell percentage was also decreased. From the third week on, however, the decrease of CFU-GM yields slowed down, while CFU-GM in the system with rhIL-2 continued to fall. In system co cultured with autologous DCs, the adherent cells contained the least percentagcs of CD34(+) cells and P210(+) cells percentage. However, the expression of BCR/ABL in CFU-GM colonies derieved from the adherent cells of DCs co-cultured had no significant difference with those from the culture without DCs. Our results suggest that co-culture of marrow cells with autologous DCs could significantly diminish the leukemic progenitors cells including both mature and primitive progenitor cells. Autologous dendritic cells might be used for ex vivo purging of CML marrow. PMID- 12578623 TI - [Detection of Cytomegalovirus Antigenemia Guides Prophylaxis of Cytomegalovirus Infection after Allogeneic Hematopoietic Stem Cell Transplantation] AB - Cytomegalovirus infection (CMV-I) and CMV related diseases (CMV-D) occurred after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) seem to be with high morbidity and mortality. This study is a retrospective analysis of the incidence of CMV infection and diseases in Allo-HSCT patients known to be CMV seropositive before transplantation. To review the efficacy of CMV pp65 antigen guided ganciclovir prophylaxis in preventing CMV infection and to search the optimal determination methods, 45 consecutive Allo-HSCT patients have been observed. Using the CMV pp65 antigenemia assay and serological analysis monitored blood samples from 23 patients with chronic myeloid leukemia (CML), 7 acute myeloblastic leukemia (AML), 6 acute lymphoblastic leukemia (ALL); other: 4 myelodysplastic syndrome (MDS), 3 non-Hodgkin's lymphoma (NHL) and 2 aplastic anemia. Forty-three patients received HLA-identical siblings transplantation and 2 from their HLA-haploidentical donors. Forty-five cases included Allo-PBPCT (38 cases), Allo-BMT (2 cases) and Allo-PBPCT + BMT 5 cases. Before transplantation, all donors/recipients have taken CMV serological detection. All donor/recipients were CMV IgG positive and one donor and one recipient with CMV IgM positive, respectively. After transplantation, all patients developed CMV antigenemia during monitoring period. Twenty-five patients developed CMV related interstitial pneumonia (CMV-IP). Patients have been followed from 6 to 28 months (median of 18 months) after transplantation. The patients who received preemptive therapy had a significantly better outcome than patients who did not received preemptive therapy. CMV related mortality was 1/29 cases in preemptive group vs. 12/16 cases in non-preemptive group. The results suggest that prompt and early institution of effective therapy with ganciclovir upon detection of CMV pp65 antigenemia, provides optimal protection against progress of CMV disease for patients undergoing Allo-HSCT. PMID- 12578624 TI - [Operating Procedure of Collection, Processing and Preservation of 3000 Units Umbilical Cord Blood in Shangdong Cord Blood Bank] AB - The experience with the umbilical cord blood (UCB) stem cells for unrelated transplantation from our 3 000 UCB storage was described. UCB, collected from closed blood bags, were mixed with hydroxyethyl starch for nucleated cell (NC) enrichment. After finishing CD34 analysis, culture of hematopoietic progenitors (CFU-GM and CFU-GEMM) assays, microbial culture, HLA Class I (A, B) serology and class II (DR) low resolution SSP typing, cord blood units are stored in the liquid nitrogen for clinical applicatoin. Cord blood contained an average of nuclear cell (NC) (1.2 +/- 0.6) x 10(9), CD34(+) cells (3.0 +/- 3.7) x 10(6), CFU GM (1.1 +/- 0.7) x 10(6) and CFU-GEMM (1.1 +/- 1.2) x 10(6) for storage and the recovery rates were 91%, 88%, 85% and 82%, respectively. The recovery rates for red blood cell and Hb were (39 +/- 9)% and (40 +/- 8)%, respectively. The storage volume was (35.1 +/- 7.1) ml in a 50 ml storage bags. The mean time from collection to processing of 15 hours (range 4 - 24 hours) had no influence on cell viability. The cell viability before processing is more than 95% and 92% after UCB thawing. The recovery rates of NC, CD34(+) cells and CFU-GM post thawing were 96%, 90% and 91%, respectively. There were no HIV antibody (HIVAb) positive in all of UCB units. For an incidence of processed samples, infection with syphilis, HBsAg, HBcAb, HCVAb, CMV, bacterial contamination and abnormal hemoglobin were 0.1%, 0.8%, 3.2%, 0.2%, 87.1%, 1.2% and 0.1%, respectively. More than 3 HLA loci matched can be found for random patients in our cord blood bank and 6 HLA loci matched have 5%. For transplantation with nucleated cell counts of > 2.7 x 10(7) cells/kg, our cord blood bank will be able to provide all of the umbilical cord blood stem cell samples for children and 50% of units can be used for some of adult recipients transplantation in the country. It is concluded that: (1) The large cord blood banking for 20 000 UCB storage is feasible in China. (2) Our system of whole procedure and methods is functionable for supplying qualified cord blood units in transplantation. (3) The volume for collection is critical to the yield of CD34(+) cells or hematopoietic progenitor cells, however cord blood NC is also important and proportional with CD34(+) cells. Only the units containing more than 8 x 10(8) cells and more than 60 ml of cord blood can be in the procession for storage. PMID- 12578625 TI - Characterization of Platelet-Associated Immunoglobulin by Flow Cytometry. AB - Measurement of platelet-associated imunoglobulin (PAIg) has frequently been applied for the diagnosis of idiopathic thrombocytopenic purpura (ITP) and other immune thrombocytopenias. In the present study, a flow cytometry (FCM) analysis has been used to detect and characterize PAIg in 47 patients with ITP and Evans' syndrome, 13 patients with non-immune thrombocytopenia, 10 patients with autoimmune hemolytic anemia (AIHA) whose platelet counts were in normal range, and 31 healthy volunteers. With FCM measurement, mean fluorescence intensity (MFI) of platelets from patients with ITP and Evans' syndrome (2.26 +/- 2.29) was significantly higher than those from non-immune thrombocytopenia (0.33 +/- 0.39), AIHA (0.17 +/- 0.07) and control subjects (0.25 +/- 0.15) (P < 0.01). Meanwhile, the percentage of positive platelets of patients with ITP and Evans' syndrome [(44.1 +/- 29.0)%] was also higher than those of non-immune thrombocytopenia [(17.5 +/- 9.4)%], AIHA [(10.7 +/- 7.5)%] and control subjects [(16.6 +/- 8.4)%] (P < 0.01). In addition, some peak shape abnormality appeared (double peaks and peak tail) in the histogram of fluorescence intensity (log) of 11 patients (23.4%) with ITP and Evans' syndrome either alone or accompanied with quantitative alteration of MFI and/or positive platelet percentage. In seven cases, the peak shape abnormality was the unique characteristic that could be detected and have never been seen in normal platelets. This phenotypic alteration perhaps reflects the existence of different platelet populations and could be of diagnostic value. Totally, the positive result of FCM measurement in patients with ITP and Evans' syndrome was 87.2%, slightly higher than 83.0% positive rate with ELISA method, without statistical difference. The correspondent rate of the results of these two analytical settings was 85.1%. This study shows that FCM assay is a rapid and sensitive method for the measurement of PAIg and seems to be suitable as a novel routine diagnostic technique of immune thrombocytopenia. PMID- 12578626 TI - [Effect of beta-Fibrinogen-455 Gene Polymorphism on Plasma Fibrinogen Levels in Patients with Ischemic Stroke] AB - In large prospective studies, plasma fibrinogen levels have been shown to be an independent risk factor of vascular disease, including ischemic stroke. Elevated plasma fibrinogen in an individual could be due to the presence of predisposing genetic and/or environmental factors, such as smoking. Of the polymorphisms studies to date, the beta-fibrinogen-455 (beta-Fg-455) G-->A substitution in the 5' flanking region is associated with the most consistent difference in plasma fibrinogen levels in both case-control studies and in selected groups of healthy individuals. In order to further elucidate the role of the beta-Fg-455 G-->A substitution in determining fibrinogen levels and susceptibility to ischemic stroke in case-control population, including 104 individuals with verified ischemic stroke and 156 healthy individuals. Turbidimetriy assays were used to measure plasma fibrinogen levels of all samples. The beta-Fg-455 G-->A mutation was identified by the polymerase chain reaction followed by restriction enzyme digestion of the amplified DNA with HaeIII. The plasma fibrinogen level in patients with ischemic stroke [(3.51 +/- 1.09) g/L] was significantly higher than that in the control [(3.08 +/- 0.71) g/L] (P < 0.01). The A-allele is associated with elevated fibrinogen levels in both patients and controls. The plasma fibrinogen levels in controls with A-allele in elder people were higher than in younger people (P < 0.05). Those with A allele in males of ischemic stroke had significantly higher plasma fibrinogen levels in smokers than in non-smokers and ex-smokers (P < 0.05), but it was not significantly difference in subjects of GG genotype (P > 0.05). Our data demonstrates an association of the beta-Fg promoter A-455 allele with higher fibrinogen levels in the general population, and suggests that the A-allele may be a susceptible predictor of ischemic stroke, particularly in aging and smoking. PMID- 12578627 TI - [Direct Injection of Plasmid DNA Expressing IL-6 Gene Improves Recovery of Thrombocytopoiesis in Irradiated Mice] AB - Hemorrhage is one of major clinical features of the patients exposed to large dose of ionizing radiation and a sudden decrease of peripheral platelet counts in hemorrhage complication may bring the patients into life-threatening situation. Cytokines had been used to improve thrombocytopoiesis in various radiation induced thrombocytopenia. Current measures for this purpose involve repeated injection of recombinant cytokines, which bring much inconvenient and agony to the patients, or gene therapy with viral vectors that could not obviate the risk of infection. This work tried to determine the possibility of gene therapy with plasmid vectors for radiation-induced hematopoietic injury. After a single intramuscular injection of plasmid hIL-6 cDNA on 6.5 Gy irradiated mice, the IL-6 level began to increase from the day 4, reached the peak value about the day 11 and maintained at a higher level on the day 28, but the hIL-6 level showed less changes in unirradiated mice. Further experiments demonstrated the IL-6 level in 7.5 Gy irradiated mice was about three times higher than that of 5.0 Gy irradiated mice and the expression of hIL-6 in vivo showed significant effect on hematopoietic recovery. Not only the platelet nadir in peripheral blood, but also the number of colony-forming cells in bone marrow rose. It is concluded that radiation could significantly enhance the gene transfer efficiency of plasmid DNA and gene therapy with plasmid vectors for treating radiation-induced hematopoietic injury might be more effective than other diseases without DNA repair. PMID- 12578628 TI - [Variant Frequency of Erythrocyte Glycophorin A in Persons Accidentally Irradiated by (60)Co Source] AB - Glycophorin A (GPA) is one of the important molecular markers in studies of somatic cell mutations. To investigate the relationship between the gamma irradiation and the frequency of GPA variation, the frequency of variant erythrocytes at the GPA locus was determined in peripheral blood of 3 subjects with accidental whole-body gamma-irradiation. The biological dose of individuals was 2.5, 2.9 and 1.9 Gy estimated by the chromosome aberration assay, respectively, and the frequency of GPA variation was 3.9, 4.3 and 4.1 times greater than that from normal controls, respectively. Our results suggest that the variant frequency of erythrocyte GPA was increased. On account of the GPA gene mutations are preserved in hematopoietic stem cells during all irradiated individual's life, the frequency of GPA variation could be used as a permanent marker for mutagenesis of radiation. PMID- 12578629 TI - [Effects of Ginsenosides Rg1 and Rb1 on Proliferation of Human Marrow Granulocyte Macrophage Progenitor Cells] AB - Ginseng is a traditional Chinese medicine which has been used in treating anemia for thousands of years. It is composed of a lot of components. The main component is total saponin of panax ginseng (TSPG), which contains more than 20 ginsenosides including Rg1, Rb1 and so on. Previous studies have reported that total saponin of panax ginseng could promote hematopoiesis by stimulating proliferation of human erythroid grogenitor cells CFU-E and BFU-E, however, it had different effects on CFU-GM reported by various laboratories. In this study, CFU-GM assay was adopted to observe the ginsenosides Rg1 and Rb1's effects on the proliferation of human marrow grannulocyte-macrophage progenitor cells. The results showed that Rg1 and Rb1 had obvious promotive effect on the proliferation of CFU-GM, and the increasing rates of colony formation were up to (70.6 +/- 6.8)% and (65.1 +/- 6.3)%, respectively. There was no inhibiting effect on CFU-GM in high concentrations of Rg1 and Rb1. It is suggested that Rg1 and Rb1 can stimulate the proliferation of human granulocyte-macrophage progentors. The results of TSPG's various effects on CFU-GM might be caused by different contents of ginsenosides in TSPG used in different laboratories. PMID- 12578630 TI - [Study on the PTEN Tumor Suppressor Gene] AB - PTEN (also designated MMAC1) gene is a new tumor suppressor gene proved to be a lipid phosphatase. The target lipid, called phosphatidylinositol-3,4,5 trisphosphate, PIP3, and it is a key component of the major cell growth control pathways, acting both to stimulate cell growth and to inhibit tumor cell proliferation blocking apoptosis. PTEN mutations have been linked to a variety of human cancers. Mutations of PTEN during tumorigenesis allow the mutated cells to grow unchecked when they should die. PMID- 12578631 TI - [A Strategy of IL-6/IL-6R System Mediates Targeted Treatment for Leukemia] AB - An inevitable trend for the development of new treatment of leukemia is to use targeted strategy. IL-6/IL-6R is important one of the cytokine receptor targeted treatment systems. Many malignant cells, including multiple myeloma, prostate carcinoma and leukemia etc., have been shown to express IL-6R. Leukemia cells, especialy in U937, TF1, KG1 cell lines highly express the high affinity IL-6R. IL 6 recombinant toxin is cytotoxic in vitro to leukemia cell expressing high affinity IL-6R; in vivo the fusion toxin can result the reduction of leukemic cell load in animal leukemia model but have no effect on normal hematopoiesis in non-leukemic animal. On the basis of these pre-clinical studies, IL-6 recombinant toxin my become novel drug for the treatment of leukemia and cancer in future. PMID- 12578632 TI - [Generation of Dendritic Cells from Human Cord Blood CD34(+) Cells and Their Activation of T Cell-Dependent Antitumor Immune Response In Vitro] AB - To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34(+) cells isolated from human cord blood by mini-MACS were cultured in a liquid culture system with rhSCF, rhGM-CSF, rhTNF-alpha and rhFL for 10 days. Then the induced cells were characterized by DC's morphological and phenotypic properties. In addition, they stimulated the proliferation of allogeneic T cells and possessed an efficient capacity for initiating T cell-dependent antitumor immune responses in vitro. It is concluded that mature DCs could be obtained from human cord blood CD34(+) cells. PMID- 12578633 TI - [Expression of nm23-H(1) mRNA in Bone Marrow Cells from Patients with Myelodysplastic Syndrome and Its Clinical Implication] AB - The purpose of this investigation was to explore the expression of nm23-H(1) gene in patients with myelodysplastic syndrome (MDS) and evaluate the relationship between nm23-H(1) expression and therapeutic outcomes. Semi-quantitative RT-PCR was used to detect the expression of nm23-H(1) mRNA in marrow mononuclear cells from 28 MDS patients and 15 normal subjects. nm23-H(1)/GAPDH ratio >/= 0.5 was believed to a positive case. The expression of nm23-H(1) was positive in 24 of 28 MDS patients, and the average level was 0.89 +/- 0.56. nm23-H(1) mRNA was negative in normal controls. The overexpression of nm23-H(1) mRNA in MDS patients could predict outcome of treatment and prognosis for MDR patients. PMID- 12578634 TI - [More on Hematopoietic Stem Cell Research and Application Updated] AB - Allogeneic hematopoietic stem cell transplantation (HSCT) is an effective and proven treatment for malignant and nonmalignant diseases. Most of the natural HSC allografts hardly show overall advantages of high engraftment, slight GVHD and rare relapse. The graft engineering including stem cell engineering to make a tailor-made graft ex vivo is promising to conquer all the risks of low engraftment, lethal GVHD and high relapse, which becomes the key program in current HSC research. To combine HSC allotransplant with gene therapy and immune therapy is the novel therapeutic strategy for malignancies, the real meaning of "cytotherapy" or "cell therapy" updated. The rapid expansion of umbilical cord blood banks (CBBs) makes the substantial increase of cord blood transplants (CBT) both possible and likely world-widely. In China, however, owing to lack of hematological pediatricians specified in HSCT and pediatric laminar-flow wards, clinical application rate of cord blood is extremely low despite of the high collection rate in the CBBs under the GMP standards. In evaluating a CBB, the release rate and the clinical efficacy of the released cord blood should be most emphasized as well as the banking quality control. In all CBBs worldwide, it is unworthy of mentioning the banking of umbilical cord blood for autologous transplant. Only one or two commercial companies in the world run it for profitable purpose to charge the donor parents regularly. It is because no autologous CBT can cure the inherited diseases and its efficacy of treating malignancies is doubtable since the cord blood is of weak immune competence against tumor and may be contaminated with autologous malignant or premalignant cells. Moreover, there is no report so far about how long the repopulating activity of cryopreserved hematopoietic stem/progenitor cells of cord blood can keep. No honest guarantee can be made about the effective quality and adequate amount of stem cells to meet the therapeutic requirement when used after a long storage. PMID- 12578635 TI - [Expression of Vascular Endothelial Growth Factor in the Bone Marrow Cells from Adult Chronic Myelogenous Leukemia] AB - Vascular endothelial growth factor (VEGF) is one of the main angiogenic cytokines and plays an important role in the development of human solid tumors. However, it is not clarified whether VEGF governs the progress of the chronic myelogenous leukemia (CML). This study is to estimate VEGF expression in the bone marrow cells from normal and adult CML patients and various leukemic cell lines. Reverse transcription-polymerase chain reaction (RT-PCR) was used for detection of VEGF mRNA. VEGF concentrations in the cell cultural supernatant and the plasma from normal and CML patient bone marrows were determined by enzyme linked immunosorbent assay (ELISA). VEGF mRNA was positive in 67 of 72 cases of bcr/abl(+) CML patient bone marrow cells (93.1%), in 5 of 10 CML patients post Allo-BMT bone marrow cells (50%), and in 6 of 10 normal bone marrow cells (60%), the expression rate of VEGF mRNA in CML patients bone marrow cells was higher than that in CML patients post Allo-BMT and normal bone marrow cells. VEGF mRNA also expressed in the HL-60, K562, CEM, KG1a, NB4, and Nalm6 cells, but not in the Jurkat cells. The mean VEGF concentration in the plasma (380.6 pg/ml) from 22 untreated CML patients was 9 folds higher than that from 9 CML patients post Allo BMT (38.0 pg/ml). The mean VEGF concentration in the cultural supernatant (499.8 pg/ml) of 17 newly diagnosed CML bone marrows was 2.5-folds higher than that in 11 normal donors (141.3 pg/ml). The CML marrow cells secrete more VEGF than normal marrow cells do. Our results suggest that the abnormality of VEGF transcription and translation expression may play an important role in the development of CML. PMID- 12578636 TI - Expression of Thymosin beta4 in Murine Bone Marrow Endothelial Cells, HL-60 Cells and Human Peripheral Blood Mononuclear Cells. AB - To investigate the relationship between the growth situation of the cells and the expression level of thymosin beta4, the specific primer of thymosin beta4 was selected to test the expression of thymosin-beta gene in murine bone marrow endothelial cells in different proliferation states and in HL-60 cells and peripheral blood mononuclear cells by RT-PCR. The results showed that the expression level of thymosin beta4 in the murine bone marrow endothelial cells with active proliferation was high, otherwise it was low. The thymosin beta4 gene expression level was higher in HL-60 cells than that in human peripheral blood mononuclear cells. It was suggested that the expression level of thymosin beta4 mRNA is closely related to cell growth. PMID- 12578637 TI - [Transfer and Expression of Recombinant Human Thrombopoietin Gene in COS-7 Cells and Mice In Vivo] AB - In order to investigate the expression of recombinant TPO gene in COS-7 cells and in vivo of the mouse model, eukaryotic expressing plasmid pcd2/TPO with human TPO cDNA was constructed with DNA recombinant techniques. The plasmid pcd2/TPO was transiently transfected into the COS-7 cells by means of lipofection, the naked pcd2/TPO plasmid was injected into the skeletal muscle of mice with electric pulses. RT-PCR and ELISA methods were used to detect the TPO expression of the transfected COS-7 cells, both showed high level expression. The MTT test showed the expressed TPO had proliferative activity to TPO-dependent cell line. High efficiency of gene transfer in transgenic mice was also observed by RT-PCR and immunohistochemical methods. The serum TPO level [(1 185 +/- 264) ng/L] in transgenic mice was quite different compared with the normal mice [(250 +/- 76) ng/L]. All these results provided solid foundations for the research of TPO gene therapy in the future. PMID- 12578638 TI - [The Application of RACE Technique to Clone the Full-Length cDNA of A Novel Leukemia Associated Gene LRP16] AB - LRP16 is a novel gene which was found in our laboratory by using methylation sensitive restriction landmark genomic scanning (RLGS) technique. In order to clone the full-length cDNA of this leukemia relapse associated gene, the method of rapid amplification of cDNA end (RACE) was employed. By optimizing some procedures of RACE method, the 5'- and 3'-untranslated region of LRP16 cDNA was successfully sequenced. Then, the full length of LRP16 cDNA and open reading frame (ORF) was constructed and was registered in GenBank. The above-mentioned procedure demonstrated RACE technique is a rapid and sensitive method for cloning unknown gene. Especially, it is very useful to cloning the 5'- and 3' untranslated region of a novel gene. PMID- 12578639 TI - [Establishment of the Cell Line U937-ASPI3K with Stable Expression of Antisense ATM/PI3K] AB - In order to provide a valuable cell model for studying the mechanism of DNA damage to kill tumor cells and enhance the curative effect, an antisense cDNA, which effectively expressed ATM gene encoding carboxyl terminal 100 kD domain fragment, was constructed, and a cell line U937-ASPI3K, which stably expressed the antisense ATM/PI3K cDNA was established through transfection and Zeocin selection. The results showed the constructed ATM/PI3K cDNA is in the opposite orientation in expression vector pZeoSV2(+) through examination of gene sequence. The cell line U937-ASPI3K was total loss of constitutive ATM protein, while the ATM abundance was not influenced in both the control cell lines U937-pZeoSV(-) and U937. The present experiment provided a practical cell model to elusidate mechanism of cell cycle checkpoints in DNA damage signal transduction pathway and search a new approach for tumor therapy. PMID- 12578640 TI - [Expression of AC133 Antigen on Acute Leukemia Cells] AB - To investigate the expression of AC133 antigen on acute leukemia cells and its clinical significance, samples of 29 patients with acute lymphocytic leukemia (ALL) and 32 patients with acute myelocytic leukemia (AML) were analyzed by flow cytometric three-color direct immunofluorescence methods. It was found that 13 of 29 ALL, 6 of 8 AML-M(0)/M(1), 4 of 16 AML-M(2) and 4 of 8 AML-M(4)/M(5) expressed AC133. The mean fluoresence intensity of AC133 in each subtype of AML-M(0)/M(1), AML-M(2) and AML-M(4)/M(5) was 227.1, 143.1 and 142.5, respectively. Among 27 AC133(+) cases (44.3%), 25 cases coexpressed CD34 (92.6%) and 13 cases coex- pressed CD117 (48.1%). In contrast to other reports, our data do not support the findings that AML-M(4)/M(5) expressed very high levels of AC133. In conclusion, AC133 antigen, coexpressed with CD34 and CD117, appears more frequently and more intensely in AML-M(0)/M(1). PMID- 12578641 TI - [New Retinoid SX-116 Induces Apoptosis of Acute Promyelocytic Leukemia Cell Line NB4] AB - In this research, the effect of novel retinoid SX-116 on acute promyelocytic leukemia cell line NB4 was studied in vitro. Cell proliferation, cell morphological characters, flow cytometry, DNA electrophoresis and RT-PCR were observational parameters. The results showed that treated with SX-116 at 10(-6) mol/L, the growth and survival of NB4 cells were markedly inhibited, morphological changes of apoptosis, including membrane blebbing, chromosome condensation and fragmentation of nuclei were observed in NB4 cells after 24 hours exposure of SX-116. Further studies showed "DNA ladder" in genomic DNA electrophoresis, as well as a typical apoptotic peak below G(1) phase presented in flow cytometry. The expression of apoptosis - related gene bcl-2 and p53 were examined. The level of bcl-2 mRNA was downregulated by 6-hour treatment of SX 116, while the gene restored to the normal level by following 12-, 24- and 48 hour exposure. However, p53 mRNA was unchanged during the treatment. The results demonstrated that SX-116 could induce apoptosis of NB4 cells while the mechanism remains to be studied. PMID- 12578642 TI - [c-myc Antisense Oligodeoxynucleotides Induce Apoptosis of Human Myeloid Leukemia Cell Line HL-60] AB - In order to study the effects of c-myc antisense phosphorothioate oligodeoxynucleotide in inducing apoptosis of HL-60 cells, the expression of c myc mRNA was determined by RT-PCR, the morphologic signs of apoptotic cells were observed by transmission electron microscopy, the proportion of apoptotic cells was detected by flow cytometry, and the DNA fragments were analysed by agarose gel electrophoresis. Results of RT-PCR showed marked decrease of c-myc mRNA expression in AspoI and II treated cells. The level of c-Myc protein was decreased 23.8% and 45.4%, respectively, in cells treated with 5 and 10 micro mol/L AspoI, and 38.4% after treatment of 10 micro mol/L AspoII. The apoptotic rates were 23.97% and 52.6% after 10 micro mol/L AspoI treatment and 28.8% and 45.19% after treatment with AspoII 10 micro mol/L for 48 and 72 hours, respectively. while apoptotic cells did not apear in control and sense oligodeoxynucleotide groups. Electron microscopy observation showed the characteritics of apoptosis. A ladder like pattern of DNA fragments was demonstrated on electrophoretogram. The results suggst that c-myc antisense oligodeoxynucleotides could be highly specific gene agents that can suppress the level of c-myc mRNA, decrease the c-Myc proteins and induce apoptosis of HL-60 cells. PMID- 12578643 TI - [Comparing the Oxygen Free Radicals and Oxygen Consumption of Leukocytes in Leukemia Patients and Healthy Persons] AB - Using the electron spin resonance (ESR) and probe technique, the species of reactive oxygen free radicals and the oxygen consumption were observed in the metabolic process of oxygen of leukocytes from leukemia patients and healthy persons. Results showed that weak ESR spectrum of hydroxyl radical ((*)OH) signal was detected in the leukemia patient's leukocytes, there was no significant difference in oxygen consumption with or without PMA stimulation; superoxide anion (O(2)) in normal leukocytes was detected, and oxygen consumption increased markedly compared to the normal respiration without PMA stimulation (P < 0.001). There was no significant difference in oxygen consumption between leukemic and normal leukocytes without PMA stimulation; however, it was higher in normal leukocyte than in leukemic leukoeytes after PMA stimulation (P < 0.001). These observations suggest that dysfunction of respiratory burst is existed in leukemic leukocytes, and their oxygen consumption is markedly lower than that of normal leukocytes. PMID- 12578644 TI - [Establishment of a Human Umbilical Vein Endothelial Cell Line Supporting Hematopoiesis In Vitro] AB - Endothelial cells are the critical cell component of hematopoietic microenvironment. For the purpose of facilitating the study on modulating effect of endothelial cells in hematopoiesis, a human umbilical vein cell line, IEC, was established. Primary human umbilical vein endothelial cells were transfected with the plasmid pSV(3neo) carried SV40 large T antigen by lipofection. The IEC cells expressed factor VIII and the UEA I-binding ratio was about 97%. The chromosome variation was existed in the cell line, with the karyotype 45, XX, -18, 18q(+). The cell line retained the proliferative capacity at least 25 passages. IEC cells stimulated the growth of granulocyte-macrophage progenitor cells in coculture of cord blood CD34(+) cells with IEC cells. It is concluded that IEC cell line possessed the biological features of endothelial cell and supported hematopoiesis in vitro. PMID- 12578645 TI - [Activation and Cytotoxicity of Bone Marrow Immunocytes by Using Various Cytokines] AB - The study aimed to explore the changes of the immunocyte quantities and cytotoxicity in bone marrow, and how many hematopoietic progenitor cells retained after the bone marrow cells were activated by different combinations of various cytokines. Bone marrow cells were divided into four groups and were cultured in vitro: (1) Control: no cytokines were added. (2) IL-2 group: bone marrow cells were activated by IL-2. (3) CD3-AK group: bone marrow cells were activated by IL 2 and CD3-McAb. (4) CIK group: IFN-gamma, IL-1, IL-2 and CD3-McAb were added. CFU GM assay and CD3 phenotype detection were performed before and after activating culture in all groups. The changes of cell quantities during culture and cytotoxicity of cultured cells were tested. CD3 positive cells markedly increased in both CD3-AK and CIK groups. The cell numbers and cytotoxicity of CD3-AK and CIK groups were higher than those of control or IL-2 group obviously after culture (P < 0.05). CFU-GM were decreased in all groups after culture and there had no significant difference among four groups. The combination of IL-2 and CD3 McAb not only stimulates the proliferation of marrow immunocytes and increases their cytotoxicity but retains enough hematopoietic progenitor cells as well. This combination of cytokines can be used to purge autologous bone marrow in vitro. PMID- 12578646 TI - [Study on Growth and Maturation of Megakaryocyte Progenitors In Vitro in Patients with Chronic Idiopathic Thrombocytopenic Purpura] AB - To investigate the growth and maturation of megakaryocyte progenitors in patients with chronic idiopathic thrombocytopenic purpura (CITP), plasma clot culture and GPIIIa monoclonal antibody and ABC immuno- histochemical kit were used to assay CFU-Meg and BFU-Meg, and the area and diameter of GPIIIa(+) cells were determined by image analyzer in 33 CITP cases. It was found that CFU-Meg and BFU-Meg were 39.27 +/- 21.44 and 5.62 +/- 3.93 per 2 x 10(5) MNC, respectively, in CITP patients, there were no significant differences with those in control group. While the area of GPIIIa(+) cells was (134.90 +/- 6.08) micro m(2) and diameter was (12.89 +/- 3.66) micro m, those were lower than those in control group. In patients with normal number of megakaryocytes on marrow smears, CFU-Meg and BFU Meg were 19.43 +/- 7.28 and 4.67 +/- 1.53, respectively, the values were lower as compared to control group. The positive correlation was showed between the total megakaryocytic colonies and the number of megakaryocytes on marrow smears, r = 0.6503, and there was no correlation with blood platelet counts and course of disease. The results suggest that there was a maturation disturbance of megakaryocyte progenitor in CITP patients and lower proliferative potential in patients with normal megakaryocyte counts on marrow smears. PMID- 12578647 TI - Therapeutic Effects of rhIL-11 on Irradiation-Induced Myelosuppression in Rhesus Monkeys. AB - The efficacy of rhIL-11 in treating thrombocytopenia and neutropenia in gamma irradiated rhesus monkeys and the variation in curative effect due to difference of administration times were studied. Healthy rhesus monkeys were exposed to 3.0 Gy (60)Co total body irradiation (TBI) to result in pancytopenia for three weeks. Treatment with rhIL-11 (30, 60 or 120 micro g.kg(-1).day(-1)) on early days (days 0 - 13 after TBI) could significantly improve the nadir of platelet count. Although the nadir of leukocyte count was not improved, the duration below 50% of its baseline value was shortened similarly to that of platelet. During the first two weeks after TBI, erythrocyte numbers of the animals treated with these doses of rhIL-11 were lower than those of the control group at first but they became higher beginning from the third week. Four monkeys were treated with rhIL-11 at 60 micro g.kg(-1).day(-1) on days 13 - 26 after TBI. The numbers of their peripheral blood cells followed the similar decrease patterns as those of control group during the first three weeks, then they were improved rapidly. By semi solid bone marrow cell culture it was demonstrated that rhIL-11 could stimulate bone marrow cells to form more CFU-Meg, CFU-Mix, CFU-E, BFU-E and CFU-GM in vitro. Histopathological observation revealed that bone marrow of the control group was devoid of hematopoietic cells and bleeding, being contrary to that of the group treated with rhIL-11, in which the cells proliferated actively. The results suggest that rhIL-11 can accelerate hematopoietic recovery of irradiated monkeys. PMID- 12578648 TI - [Study of Low Dose Irradiation Enhancing the Effect of Chemotherapeutic Agents against Leukemia and Its Mechanism] AB - For effectively enhancing the anti-leukemia effect of chemotherapeutic agents, and meanwhile decreasing the side effect of these agents, the study has been made to explore the synergistic effect of low dose irradiation (LDI) combined with Ara C on murine leukemia and its mechanism. Firstly, an optimal scheme of low dose total body irradiation combined with Ara-C was established in L615 leukemia (T lymphocytic leukemia) mouse model. The machanism of the enhancing effect was explored by patho-morphological observation, examination of residual leukemia cells, the expression of GM-CSF on the surface of marrow stromal cells and in the bone marrow cultural supernatants. The results showed that the optimal scheme was 300 cGy irradiation at 4 days after inoculation of leukemic cells followed by Ara C 30 mg/kg x 3 days in an interval of 1, 2 or 3 days after irradiation. The mean survival time of the L615 leukemia mice in LDI + Ara-C combined treatment groups was longer than that of control groups. The percentage of long-term survival mice (> 30 days) was the highest (58% - 72%), too. 17% of the mice were be cured. The numbers of blood leukocytes and marrow nucleated cells were transiently decreased in combined treatment group, and then recovered rapidly. Slight myelosuppression and marrow sinus dilation and congestion were seen after 300 cGy irradiation. The expression of GM-CSF either on the stromal cells or in marrow cultural supernatant after irradiation increased strikingly (P < 0.05). Therefore, LDI combined with Ara-C possesses synergistic effect. The mechanism is possibly related to three facts: LDI could increase the permeability of bone marrow sinus; LDI could promote marrow stromal cells to produce some cytokines (such as GM-CSF, etc.) which drive leukemia cells into cell cycle to make the cells more sensitive to chemotherapeutic agents; and LDI could augment Ara-C-induced cytotoxicity through the mechanism of apoptosis. PMID- 12578649 TI - [Therapeutic Effects of Lishengsu on Chemotherapy Induced Leukopenia in Patients with Malignant Tumor] AB - To observe therapeutic effects of Lishengsu (recombinant human granulocyte colony stimulating factor, rhG-CSF) on leukopenia induced by chemotherapy, 40 patients with malignant tumor included in the clinical observation were randomly divided into group A and B (20 patients each). Each patient was received two cycles of chemotherapy. In group A Lishengsu was administered in the first cycle and none in the second cycle, while in group B Lishengsu was given in the second cycle, but not in the first cycle. Lishengsu was subcutaneoasly injected for 5 - 7 days at dosage of 75 micro g or 150 micro g per day 24 - 48 hours after completion of chemotherapy. The results showed that Lishengsu remarkably alleviated the degree of leukopenia and neutropenia, and lower the incidence of infection. The curative effect of Lishengsu is reliable with slight side-effects, and it might assist to completion of chemotherapy in tumor patiants. PMID- 12578650 TI - [Short Course High Dose Mitoxantrone with Cytarabine for Treatment of Refractory and Relapsed Acute Leukemia] AB - Although chemotherapy can achieve a high rate of disease remission in patients with newly diagnosed acute leukemia, patients with recurrent or refractory leukemia generally have a poorer rate of response. This study was designed to assess the utility of high-dose mitoxantrone and intermediate-dose cytarabine in the treatment of patients with relapsed or refractory acute leukemia. Thirty patients with relapsed or refractory acute leukemia were treated with mitoxantrone at a total dose of 40 mg/m(2) intravenously and cytarabine 1 - 1.5 g/m(2) over 3 hours once daily for five doses. Twenty-six of thirty patients achieved complete response (CR 87%) and one achieved partial response (PR 3.3%). No patient died during the induction course. The main toxic effect was hematopoietic suppression that was clinically acceptable. The median time needed for CR was around 30 days. The median disease-free and overall survival times were 3.5 and 6 months, respectively. The results demonstrate that short course high-dose mitoxantrone with cytarabine is associated with a trend toward a higher CR rate and therefore, it should be an effective antileukemic regimen for the treatment of refractory and relapsed acute leukemia. PMID- 12578652 TI - [Human Mismatch Repair System in Hematologic Malignancies] AB - The human mismatch repair (hMMR) system consists of members of three super families, namely MutH, MutL and MutS. They are able to correct the error of nucleotide incorporation, increase the fidelity of DNA replication, reduce spontaneous mutation, and maintain the stability of microsatellite loci and genome. The dysfunction of MMR may induce to carcinogenesis. The relation of MMR and hematologic malignancies is presented in the article. PMID- 12578651 TI - [A Method of Crossmatching for Small Amount of Blood] AB - In order to search into the feasibility for method of crossmatching with small amount of anticoagulant blood from infant or senile patients, 76 crossmatching tests were performed with ABO compatible blood. The blood samples with ABO incompatible were divided into 6 groups: A and B, A and O, A and AB, B and O, B and AB, or O and AB, and 3 crossmatching tests were done for each group. The results showed the incomplete antibodies were examined in 1 case of ABO compatible crossmatching test, and no transfusion reactions were observed after transfusion with 76 blood matching tests. Agglutination phenomena were appeared in all of the ABO incompatible matchings. The observations suggest that the small amount of anticoagulant blood from recipient patients is feasible for crossmatching. PMID- 12578653 TI - [Non-Myeloablative Allogeneic Hematopoietic Stem Cell Transplantation-New Opinion about Hematopoietic Stem Cell Transplantation] AB - Non-myeloablative allogeneic hematopoietic stem cell transplantation (NMAT) is brought forward recent years. It is a modified transplant technology. NMAT treats malignant disease by graft versus tumor effect of chimerism after transplantation. The difference between this transplantation and standard allogeneic stem cell transplantation is the conditioning regimen. Non myeloablative allogeneic stem cell transplantation increased the indication of allogeneic stem cell transplantation. This article focused on the background of this transplantation, the results of animal experimental and clinical research, the advantages and shortages of this transplantation. PMID- 12578654 TI - [Quality Control in Umbilical Cord Blood Bank] AB - Recent clinical reports have demonstrated that the use of umbilical cord blood (UCB) opened a new source of stem cell for hematopoietic stem cell transplantation, leading to the development of cord blood banks world-wide. Prior to the large scale construction of UCB banks, quality control must be performed for health care providers and manufactures. With increasingly stringent regulatory requirement in blood industry, quality control is playing an important role in the operation of blood centers and stem cell laboratories. Reviewed the lectures in the biology of UCB and UCB banks published in recent years, our experiences were discussed in setting up Shandong blood bank to define process variables associated with the collection of UCB, to determine and optimize the procedures and materials used, to ascertain how UCB can be processed in clean room as mononucleated cell preparations, and to analyze using of long-term storage of UCB in research and clinic in the future. Our conclusions are: (1) the establishment of UCB banks for use in transplantation appears to be easy, effective and particularly suitable approach in China under cGMP conditions; (2) the procedures for volume reduction by closed and semi-automated blood processing system, SSP HLA typing, biocode and local computer net, microbiological tests and the 50 ml cryobags for storage constitute a cost efficient system for large-scale UCB banking; (3) the average of 60 ml UCB collection may contain sufficent marrow repopulating cells for children and most of adult recipients; and (4) hematopoietic stem and progenitor cells in cord blood have a more potent proliferative ability than those derived from bone marrow in cell expansion potentials. PMID- 12578655 TI - [Human Bone Marrow Mesenchymal Stem Cells Differentiate into Neuron-Like Cells In Vitro] AB - Recent reports have clearly demonstrated that bone marrow cells can be differentiated into neurons, suggesting the existence of cells with the differentiation capacity in the bone marrow cell population. It is well known that hematopoietic stem cells as well as mesenchymal stem cells (MSCs) can be transplanted and therefore, alternative of them might contribute to the process. In the present study it was addressed whether marrow MSCs could be coaxed into neuron-specific antigen bearing cells and if so, whether the differentiated cells possess the cytochemical features seen in neurons. The report here showed that high concentration of 2-mercaptoethanol (2-ME) could induce some of the MSCs into neuron-like cells expressing neurofilament (NF) and neuron specific enolase (NSE). The neuron-like cells were alkaline phosphotase positive while the others MSCs were kept negative. Cells treated with 2-ME were positive for alpha naphthylacetate esterase and glycogen and negative for acetylchonlinesterase, which were similar with the results seen in untreated cells. Furthermore, Nissel body was not observed in treated cells shown by toluidine blue staining. Therefore, it is likely that the cells described here seem not belong to the neuronal lineage. These findings, however, reveal that human MSCs could alter their committed fates under some circumstances. PMID- 12578656 TI - [Fetal Bone Marrow Stromal Cells in Cooperation with Exogenous Cytokines Support In Vitro Expansion of Cord Blood CD34(+) Cells] AB - To elucidate the role of fetal bone marrow stromal cells (FBMSC) in cooperation with exogenous cytokines in supporting the in vitro expansion of cord blood CD34(+) cells which were purified by negative immunomagnetic selection, FBMSCs were cultured with different combinations of cytokines including SCF, IL-3, IL-6, FL, G-CSF and EPO in a 14-day liquid culture system. The results showed FBMSC plus SCF, IL-3, IL-6, FL and EPO was the most effective combination which increased total nucleated cells, CFU-GM, BFU-E and CD34(+) cells by (692.4 +/- 52.7) fold, (237.1 +/- 106.6) fold, (114.8 +/- 32.8) fold and (25.3 +/- 10.1) fold, respectively. Our studies indicated that fetal bone marrow stromal cells combined with above-mentioned cytokines can efficiently expand cord blood CD34(+) cells. PMID- 12578658 TI - [Using RevTet-On System to Control Thrombopoietin Gene Expression] AB - For the purpose of thrombocytopenia gene therapy, recombinant RevTet-On and pRevTRE/TPO retrovirus were packaged and transfected to NIH3T3 after selected with G418 and hygromycin, the two inserting recombinant retrovirus cell strain RevTet-On3T3/TPO were established. TPO expression was controlled and regulated by doxycydine (Dox). After using Dox to control the expression of TPO in RevTet On3T3/TPO cells, the result showed that when Dox is added to the RevTet-On3T3/TPO cells, cell populations expressed TPO highly in the presence of 2 mg/L of Dox, and lowly in the absence of Dox. By using the RevTet-On gene expression system (the retrovirus vector RevTet-On regulation system to control the expression gene by Dox), it could modulate the expression of multiple genes by tetracyline and its derivatives. This system maybe provides a safe and efficacient way for the thrombocy to penia gene therapy. PMID- 12578657 TI - [Effect of Shark Chondroitin on T Lymphocyte Subsets of Cancer Patients] AB - The objective of the study is to find out the effect of shark chondroitin on T lymphocyte subsets in cancer patients. Patients were divided into two groups. One group was treated with chemotherapy alone, and the other group was treated with chemotherapy plus shark chondroitin. Using immunofluorescence technique, T lymphocyte subsets in the peripheral blood were determined in two groups before and after chemotherapy. The results showed that CD4(+)/CD8(+) ratio increased in the patients received shark chondroitin. In the chemotherapy group, CD3(+) had no change, but CD4(+) decreased while CD8(+) increased significantly. The results suggest that shark chondroitin could enhance immune function in cancer patients, especially during chemotherapy. PMID- 12578659 TI - Cryopreservation of Dendritic Cells Grown in Vitro from Monocytes for Their Future Clinical Use. AB - Dendritic cells are professional antigen presenting cells which are being used as adjuvants in tumor vaccination trials. Most clinical protocols currently include 4 to 10 weekly infusions of doses > 10(6) cells, each inoculum coming from a simple culture of blood monocytes. In the present study, several millions of dendritic cells from a single leukapheresis were produced; monocytes were isolated by elutriation and then cultured in Teflon bags in presence of 800 U/ml GM-CSF + 100 micro g/ml IL-13 + 10% fetal calf serum (FCS). The dendritic cells from this single batch were aliquoted in many doses for potential multiple infusions and cryopreserved in 10% DMSO + 2% human albumin in Teflon-kapton Fresenius bags either at -1 degrees C/min using a controlled rate freezer, or putting the bags directly in a -80 degrees C mechanical freezer without controlling the temperature rate. Six experiments were carried out. After one month of cryopreservation, the cells were thawed in a 40 degrees C water bath. Before and after freezing, cells were evaluated for immunophenotype (CD1a, CD14, CD40, CD80, CD83, CD86, CD54, CD58, CD16, CD32, CD64 and HLA-DR) and for their capacity to stimulate allogenic (MLR) or autologous (antigen presentation tests) lymphocytes. The results demonstrated that the mean recovery rates after freezing in liquid nitrogen or at -80 degrees C were (67 +/- 14)% and (71 +/- 13)% respectively, without any significant difference between the two techniques. The immunophenotype was not modified by the freezing-thawing procedure, as well as the lymphocyte stimulating capacities. In conclusion, our study showed that substantial numbers of functional DCs can be derived from peripheral blood monocytes using Teflon bags. DCs can be cryopreserved in a good laboratory practice setting for further clinical trials with an acceptable loss of cells and without modification of their functions. PMID- 12578660 TI - [Cloning and Expression of Various Deletants of Gsalpha Gene in Escherichia Coli] AB - Gsalpha gene mutation has been discovered in some human tumors. In our previous studies, three novel deletants of Gsalpha gene, Gsalpha L-1(500 bp), Gsalpha L 2(300 bp), and Gsalpha L-3(200 bp), and wild type Gsalpha-4(1 200 bp) were found in human leukemia cell lines and detected in leukemic cells from patients with acute leukemia. To investigate the construction, function and biological significance of the deletants, the plasmids of Gsalpha L-1, Gsalpha L-2 and wild Gsalpha-4 were transformed into E. coli DH5, amplified by PCR, and cloned in expression vector pET22b(+), and then transformed into E. coli, respectively. As a result, higher levels of expression of three recombinants were obtained in form of inclusion bodies. The results suggested that these Gsalpha isoforms have an open reading frame of gene and can be expressed in vitro. The data lay a foundation to study the relation of Gsalpha gene to leukemogenesis. PMID- 12578661 TI - [The Insertional Mutation of Transcription Factor GATA-2 Gene in Acute Non Lymphocytic Leukemia] AB - To investigate the expression and mutation of transcription factor GATA-2 gene in acute non-lymphocytic leukemia (ANLL), the GATA-2 gene were amplified in peripheral blood mononuclear cells from 85 cases with ANLL using RT-PCR, and the PCR products were further analyzed by single strand conformation polymorphism (SSCP) and sequencing. The results showed that GATA-2 gene was expressed in the most cases with ANLL (89.4%). One larger band was observed by RT-PCR in a case with M(1) subtype. Nucleotide sequence analysis revealed a insertion of 18 nucleotides at position 1 288 bp of the GATA-2 gene cDNA, thereby leading to 6 additional amino acids in second zinc finger complex of GATA-2, and it includes a cysteine which can bind to zinc ion. The insertional mutation may change the function of GATA-2 transcription factor. To our knowledge this is the first description of insertional mutation in the GATA-2 gene. PMID- 12578662 TI - [Construction and Expression of Retroviral Vector Encoding Enhanced Green Fluorescent Protein] AB - Retroviral vectors are wildly used as vehicles for gene transfer into hematopoietic cells based on its potency for efficient gene delivery and integration of transgene in host genome. The development of better transduction protocols depends on gene markers that allow a rapid detection and effective selection of genetically transduced cells. In this study, the enhanced green fluorescent protein (EGFP), a gene that is optimized for detection and expression in mammalian cells, was firstly amplified and cloned by high-fidelity PCR. The vector LGSN carrying EGFP gene was then constructed and the retroviral producer cell lines that yield high titers of LGSN vector in supernatants were developed by liposome-mediated transfection in combination with cross infection. Both GP + envAm12 murine fibroblasts and K562 leukemic cells transduced with EGFP virus demonstrated a stable green fluorescence signal readily detectable by flow cytometry or fluorescence microscopy in up to 97% and 86% of examined cells, respectively. The integration of LGSN provirus in transduced cells was confirmed by PCR analysis. These results indicate EGFP is a suitable reporter molecule for gene transfer and expression in hematopoietic cells. Therefore, the bright and long-term expression of EGFP in living cells will advance the study of gene therapy in vitro and in vivo, particularly for human applications. PMID- 12578663 TI - [Study on Methylation of p15(INK4B) Gene in Acute Myeloid Leukemia and Chronic Myeloid Leukemia] AB - In order to explore the role of p15(INK4B) gene with highly methylated CpG island in the pathogenesis of leukemia, the expression levels of p15(INK4B) gene was detected in patients with AML and CML. Methylation-specific PCR (MSP) assay was employed in the experiments. The methylation incidence was 83.9% (26/31) in AML and 0% (0/28) in CML. The results showed that methylation of p15(INK4B) gene was the one of the major ways for inactivation of the gene, and the methylation could be appeared in clinical development of the disease and patients condition worsened. Methylation of p15(INK4B) did not occur and its function probably is normal in CML. PMID- 12578664 TI - [Detection of High Methylation of p15(INK4B) and p16(INK4A) Genes in Multiple Myeloma] AB - To study the action, characteristics and expression of high methylation of p15(INK4B) and p16(INK4A) genes in multiple myeloma (MM), the sensitive methylation specific PCR method was employed to detect the hypermethylation of p15(INK4B) and p16(INK4A) in 24 patients with MM. Results showed that the methylation incidence of p15(INK4B) and p16(INK4A) genes were 70.8% (17/24) and 58.3% (14/24) in the MM patients, with the products of 148 bp and 150 bp fragments, respectively. The methylation of p15(INK4B) and p16(INK4A) genes were simultaneously happened in MM patients of plasmocytoma type with two cases at II phase and two cases at III phase. The simultaneous non-methylation of p15(INK4B) and p16(INK4A) genes were founded in five cases of MM patients, all of the tumor cells were of small plasmocyte type with mature differention. Conclusion suggested that there were high incidence of methylation of p15(INK4B) and p16(INK4A) genes in patients with MM. Hypermethylation can be detected in the early stage of disease, which was associated with its progress. It indicated a bad prognosis when methylation happended simultaneously in the two genes. Methylation of p15(INK4B) and p16(INK4A) genes may be related to the pathogeny of MM. PMID- 12578665 TI - [Effects of rhGM-CSF on Apoptosis of HL-60 Cells Induced with VP-16 Treatment] AB - In order to investigate the effects of rhGM-CSF on apoptosis of HL-60 cells induced with VP-16 treatment, HL-60 cells were first incubated with rhGM-CSF before they were treated with VP-16. The apoptosis processes and the changes in apoptosis related gene bcl-2 and fas expression were observed. The morphological and ultrastructural changes were observed under optics microscope and electromicroscope. DNA fragmentation were detected by agarose gel electrophoresis, the apoptotic rate, bcl-2 and fas expression with flow cytometry. Our results showed that rhGM-CSF inhibited the apoptosis of HL-60 cells induced with VP-16 treatment, which enhanced the down-regulation of bcl-2 expression, but inhibited the up-regulation of fas expression. So it suggested that rhGM-CSF can decrease the sensitivity of HL-60 cells to VP-16, probably by down-regulation of fas expression. PMID- 12578666 TI - [Clinical and Experimental Analysis of Myeloid Antigen Positive Childhood Acute Lymphoblastic Leukemia] AB - The purpose of this investigation was to study the myeloid antigen expression and its relationship with clinical and biological features in children with acute lymphoblastic leukemia (ALL). A panel of lineage-associated monoclonal antibodies and indirect immunofluorescence technique were used to analyse the immunophenotype in 85 previously untreated cases. The results showed that twenty one point two percent of patients expressed myeloid antigens (My(+)), and CD13 and CD33 were frequently involved. The incidence of myeloid antigen expression had no statistical difference between T and B ALL or between ALL-L(1) and ALL L(2) (P > 0.05). Myeloid antigens were expressed in three of seven T/B mixed ALL cases. There was no significant difference in clinical and biological features and chromosome abnormality between My(+) and My(-) cases (P > 0.25). There was no significant difference in complete remission rate and relapse rate between My(+) and My(-) cases (P > 0.05). The conclusion suggested that myeloid antigen expression was not associated with complete remission rate in childhood ALL. T/B mixed ALL more frequently showed expression of myeloid antigens and had an unfavorable prognosis. PMID- 12578667 TI - [The Role of Calcium Ion in Apotopsis of HL-60 Cells Induced by VP-16] AB - To study the significance of calcium in the apoptosis of HL-60 cells induced by VP-16, the technology of flow cytometry, confocal laser scanning microscopy and Western blot were used. The results showed that VP-16 could induced the apoptosis of HL-60 cells and transient increase of intracellular calcium concentration; EGTA [ethylene glycol-bis(2-aminoethyl)-N,N,N',N'-tetraacetic acid], that could combine the extracellular calcium, did not prevent the apoptosis of HL-60 cells. BAPTA-AM [1,2-bis(2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxy-methyl) ester], however, a chelating agent of intracellular calcium ions, could prevent apoptosis and the release of cytochrome C from HL-60 cells. It was concluded that the calcium plays an important role in apoptosis and the release of cytochrome C. PMID- 12578668 TI - [Homoharringtonine Induces Apoptosis of K562 Cells through Inhibition of P210(bcr/abl)] AB - In this study, the underlying antileukemic mechanisms of homoharringtonine (HHT) were investigated. K562 cell line was used to observe the effects of HHT on the induction of apoptosis and on the expression of the specific chimeric protein P210(bcr/abl), as evaluated by flow cytometric annexin V-PI dual labeling technique and Western blot. The results showed that HHT induced K562 cells to apoptotic death at the concentrations of 5 - 20 ng/ml, and some of the cells became necrotic when exposed to a higher concentration. The amount of P210(bcr/abl) oncoprotein was decreased by approximately 70% when the cells were exposed to HHT for 48 hours, however, that of its partner P145(c-abl) proto oncoprotein was not affected. It is clear from the study that HHT is an inhibitor of P210(bcr/abl) oncoprotein and therefore promotes the apoptosis of CML cells. It could be promising that HHT be used extensively in the chemotherapy of patients with CML. PMID- 12578669 TI - [Effect of Arsenic Trioxide on Myelomonocytic Progenitor Cells in Patients with Myelodysplastic Syndrome in Vitro] AB - The Effect of arsenic trioxide (As(2)O(3)) on myelomonocytic progenitor cells of patients with myelodysplastic syndrome was studied. Bone marrow CFU-GM was assayed in the agar semi-solid culture, and the bone marrow cells were incubated in liquid culture and the expressions of CD33, CD15 and bcl-2 on the marrow cells were detected by APAAP method in 24 patients. The suppressive effects of As(2)O(3) to CFU-GM were increased with the rise of As(2)O(3) concentrations. The suppression of As(2)O(3) (0.388 micro mol/L) to cluster formation was stronger than to colony formation, the suppressive rates were 70.78% vs 34.05% in low-risk group, and 86.76% vs 65.86% in high-risk group (P < 0.01), respectively. 0.194 micro mol/L of As(2)O(3) decreased clusters and increased colonies in low-risk group, but decreased clusters and did not change colonies in high-risk group. High concentration (1.94 micro mol/L) of As(2)O(3) downregulated the expression rate of CD33 and CD15 in both groups, and low concentration (0.194 micro mol/L) downregulated the expression rate of CD33 and upregulated the expression rate of CD15 in low-risk group, but decreased expression of CD33 and did not alter CD15 in high-risk group. At the same time, the high concentration of As(2)O(3) downregulated expression of bcl-2 and resulted in karyopyknosis and cytoplasm condensation; low concentration generated similar effect on expression of bcl-2 and cell morphology in high-risk group, but did not affect in low-risk. It is concluded that As(2)O(3) suppressed myelopoiesis and impelled myelomonocytic cells to apoptosis, low concentration of As(2)O(3) induced the proliferation and differentiation of myelomonocytic cells in low-risk group, however, suppressed the growth of myelomonocytic cells and accelerated the cells apoptosis in high risk group. PMID- 12578670 TI - [Mobilization of Peripheral Blood Stem Cells with High Dose Cyclophosphamide Combination Chemotherapy and G-CSF in Breast Cancer Patients] AB - To evaluate the effect of mobilization of peripheral blood stem cells (PBSC) with high dose cyclophosphamide combination chemotherapy and G-CSF in breast cancer patients, a new mobilization protocol was designed on the basis of standard combination chemotherapy regimen, in which the dose of cyclophosphamide was raised to 2 to 4 times, and G-CSF began to be used at the dose of 150 micro g twice everyday when white blood cell (WBC) decreased below 1.0 x 10(9)/L. PBSC collection was performed while WBC increased over 5.0 x 10(9)/L during bone marrow recovering. The PBSC mobilization protocol was completed in 10 patients, the median nadir of WBC was 0.8 (0.4 - 1.0) x 10(9)/L, the median time of PBSC collection was 2 (2 - 4), the median number of collected CD34(+) cells was 6.43 (1.99 - 8.75) x 10(6)/kg. The results showed that the protocol, high dose cyclophosphamide combination chemotherapy, was an optimal PBSC mobilization regimen in breast cancer patients. PMID- 12578671 TI - [In Vitro Expansion of Bone Marrow Stem/Progenitor Cells by Human Placental Cell Free Suspension] AB - In order to explore the expansive effect of human placental cell-free suspension (HPCFS) on bone marrow hematopoietic stem/progenitor cells, and to compare the effect of HPFCS with some cytokines and their combination, human marrow CFU-GM, CFU-GEMM and BFU-E were assayed in a semisolid methyl cellulose culture system using HPCFS, IL-3, GM-CSF, and IL-3 + IL-6 + GM-CSF + EPO as colony stimulating factors, respectively. The results showed that HPCFS stimulated the growth of CFU GM, CFU-GEMM and BFU-E and the optimal concentrations for stimulating effect were 200 - 300 micro g protein/L, and the yield of 3 kinds of colony in HPCFS group was higher than that in IL-3, GM-CSF and IL-3 + IL-6 + GM-CSF + EPO groups. The expansive effect of HPCFS on marrow progenitors was superior to IL-3, GM-CSF and IL-3 + IL-6 + GM-CSF + EPO. Human placental cell-free suspension contained a variety of cytokines to stimulate proliferation of hematopoietic stem/progenitor cells, and it could be used as an efficacious and inexpensive agent to expand hematopoietic stem/progenitor cells in vitro. PMID- 12578672 TI - [Standardization and Quality Control in Flow Cytometric Enumeration of CD34(+) cells] AB - The enumeration of CD34(+) cells by flow cytometry is commonly employed to assess hematopoietic stem/progenitor cells in cord blood, peripheral blood and apheresis products. Interlaboratory variation of CD34(+) cells enumeration is exceedingly large. Factors contributing to those variation, currently established flow cytometry protocols and comparison between these protocols were reviewed. CD34(+) cells in 45 cord blood samples, 12 normal bone marrows and 4 apheresis products were also enumerated in our laboratory by using ProCUNT kit, and results showed that the ProCUNT is highly standardizaed and could assist in reducing interlaboratory variation of CD34(+) cells. PMID- 12578673 TI - [WT1 Gene and Leukemia] AB - WT1 is classified as a tumor suppressor gene that can repress transcription of many growth factor and growth factor receptor genes. However, WT1 is highly expressed in human leukemic cells and overexpression of WT1 correlates with poor prognosis. Furthermore, suppression of WT1 expression by the WT1 anti-sense oligonucleotide inhibits proliferation and induces apoptosis of leukemic cells. Constitutive WT1 expression in myeloid progenitor cells by the transfection of the wild-type WT1 gene promoted their proliferative response to G-CSF, but inhibited their differentiation. These findings suggested that WT1 gene might perform an oncogenic function in hematopoietic progenitor cells. PMID- 12578674 TI - [Roles of Cytokines in Acute Graft-Versus-Host Disease] AB - Acute graft-versus-host disease (aGVHD) is the major complication of allogeneic stem cell transplantation (allo-HSCT), significantly limits the application of the therapy. Current evidence suggests that dysregulated cytokine production is responsible for many manifestations of aGVHD. The mechanisms have been most clearly delineated in mouse models, detailed analysis of human tissue is required. Monitoring serum levels of cytokine sIL-2R, TNF-alpha and IFN-gamma after transplantation or cytokine gene expression before transplantation can predict prognossis of aGVHD. GVHD have graft-versus-leukemia (GVL) effect, and GVL can be seperated from GVHD. IL-2, IL-12, IL-11, KGF and G-CSF could possess the roles of reducing GVHD while preserving GVL. PMID- 12578675 TI - [The Function of Dendritic Cells Derived from Chronic Myeloid Leukemia] AB - The purpose of this study was to investigate the function of dendritic cells derived from chronic myeloid leukemia (CML-DC). Mononuclear cells were prepared from bone marrow and peripheral blood of 24 patients with CML, and the DCs were obtained by incubation of MNCs with media containing GM-CSF, IL-4 and TNF-alpha. The phenotype of CML-DCs was identified by flow cytometry. FITC-dextran uptake, (3)H-TdR incorporation or MTT assay and lactate dehydrogenase release assay were used to detect uptake of exogenous antigen in immature DCs, the antigen presenting ability in mature DCs and specific cytotoxicity of CTL to leukemic cells, respectively. The DCs with high expression of CD1a, CD86, CD80, HLA-DR, CD54 and CD4 were obtained from marrow and blood of patients with CML. The uptake of FITC-DX was observed in early DCs. There was a potent stimulation to allo-MLR in DCs cultured for 7 - 10 days, and a lightly lower stimulation to auto-MLR. CML DCs can induce the generation of specific cytotoxic T cells. These results suggest that CML-DCs are functional DCs with the ability to induce anti-leukemia effect. PMID- 12578676 TI - Cleavage of Bcl-2 Protein by Activated Caspase-3 Is Associated with Inactivation of Lyn(p53/56) Kinase Activity in Human M-07e Leukemic Cells during Apoptosis. AB - The growth of M-07e human megakaryocytic leukemia cells is strictly dependent on GM-CSF. In M-07e cells, the GM-CSF receptor (GM-CSF R) is composed of two subunits: a low affinity alpha subunit and a phosphorylated beta subunit, which is constitutively linked to lyn(53/56) protein tyrosine kinase. In this study, The role of lyn kinase in regulating TGF-beta 1-induced apoptosis in M-07e cells was examined. The removal of rhGM-CSF from the culture medium resulted in down regulation of lyn kinase activity, followed by growth inhibition and programmed cell death. Apoptosis of M-07e cells was accompanied with a massive cleavage of Bcl-2 and Bax proteins into shortened fragments with molecular mass of 22 kD and 18 kD, respectively. Using specific inhibitors, the cleavage of Bcl-2, but not Bax, was found to be processed through activated caspase-3 (CPP32), which is abundantly expressed in M-07e cells. TGF-beta 1 inhibited rhGM-CSF-stimulated cell growth and promoted apoptosis in M-07e cells with a pattern identical to that induced by rhGM-CSF depletion, which included massive cleavage of both Bcl-2 and Bax proteins and inactivation of lyn kinase activity. TGF-beta 1 did not affect the levels of lyn protein or the beta-subunit, neither did it block the interaction between these two components. Also, TGF-beta 1 treatment did not diminish the expression of the alpha subunit in M-07e cells. Our results showed that TGF-beta 1 inhibits cell proliferation and promotes apoptosis in M-07e cells by inactivating the GM-CSF R-associated lyn kinase activity. Further, This study showed that Bcl-2 cleavage by activated CPP32 is a naturally occurring event associated with apoptosis, which is under the regulation of lyn kinase activation. PMID- 12578677 TI - [Expression of GATA-2 Gene and Immunoglobulin Heavy Chain Germline Gene C( micro ) in Leukemia Cells and Its Significance] AB - To investigate the significance of GATA-2 and immunoglobulin heavy chain germline gene C( micro ) (IgH germline gene C( micro )) expression and coexpression in various leukemia cells, GATA-2 and IgH germline gene C( micro ) mRNA in bone marrow and peripheral blood cells from 63 leukemia patients were detected by reverse transcription-polymerase chain reaction (RT-PCR). No GATA-2 or IgH germline gene C( micro ) mRNA were detected in normal bone marrow and peripheral blood. GATA-2 mRNA were be detected in 91.3% patients with acute myeloid leukemia (AML), 75% patients with acute lymphoblastic leukemia (ALL) as well as 83.3% patients with chronic myeloid leukemia (CML-CP); IgH germline gene C( micro ) mRNA were be identified in 47.8% AML, 41.6% ALL, as well as 5.6% CML-CP. All patients with CML-AP and CML-BC expressed GATA-2 mRNA and partly expressed IgH germline gene C( micro ) mRNA. 47.8% AML and 41.6% ALL patients coexpressed GATA 2 and IgH germline gene C( micro ) mRNA. GATA-2(+) IgH germline gene C( micro )(+) cells of AML and ALL were mainly HLA-DR positive. As aberration of the transcription factors, GATA-2 and germline IgH germline gene C( micro ) gene might been linked to leukemogenesis. Various expression of GATA-2 and germline IgH germline gene C( micro ) gene in leukemia might correlated with the heterogeneous differentiation level of leukemia cells. The fact that leukemia with GATA-2(+) IgH germline gene C( micro )(+) coexpression indicated multilineage impairment of hematopoietic cells. PMID- 12578678 TI - [Application of Metaphase-Fluorescence in Situ Hybridization to the Diagnosis of Acute Promyelocytic Leukemia and the Detection of Minimal Residual Disease] AB - To investigate the value of metaphase-fluorescence in situ hybridization (M-FISH) in the diagnosis of acute promyelocytic leukemia (APL) and the detection of its minimal residual disease (MRD), 10 cases of untreated acute myeloid leukemia (AML) (de novo APL 5 cases, relapsed APL 3 cases, AML-M(1) and AML-M(5) one case each) diagnosis by cell morphology at presentation and 10 cases of APL after complete remission (CR) were studied by M-FISH using a whole chromosome 17 painting probe labeled by digoxigenin and the results were compared with that of conventional cytogenetic examination and reverse transcription-polymerase chain reaction (RT-PCR). Among 10 untreated AML cases, 7 had positive M-FISH results, of whom 4 had t (15;17) translocation, 3 had normal karyotype. Six of them had PML/RARalpha fusion transcript except one, in whom RT-PCR did not be performed; 3 had negative M-FISH results, of whom one had del (2q) x 2 abnormalities, who was RT-PCR-positive for PML/RARalpha fusion transcript; one had complex karyotype abnormalities, whose RT-PCR was negative for PML/RARalpha fusion transcript; one had t (9;22) translocation, whose RT-PCR was negative for PML/RARalpha fusion transcript, but positive for BCR/ABL fusion transcript. Thus the diagnosis of AML M(3) was revised as AML-M(2) for the latter two cases. 10 APL cases after CR had normal karyotype, but 12/15 M-FISH assays detected 1 - 5 t (15;17) positive cells in 9 of them. This finding is compatible with the results of RT-PCR assays. Leukemia relapse was seen in one case, and two positive M-FISH results were appeared in the 2 assays at a 13 months' interval. This study suggests that M FISH had important practical value in the diagnosis of APL and the detection of MRD, and that it is less sensitive than RT-PCR, however, it seems to be more potential for prediction of the relapse of leukemia due to its capacity of detecting quantitatively the chromosome translocation in proliferative cells. PMID- 12578679 TI - [Preliminary Study on the Effects of Novel Retinoids SX-115 and CHU-012 on NB4 Cell Line] AB - In this research, the effects of two new retinoids, SX-115 and CHU-012, on promyelocytic leukemia cell line NB4 were studied in vitro. Cell proliferation, cell morphologic characters, cell cycle kinetics, reduction ability of NBT, differentiation antigens, immunofluorescence staining and RT-PCR were adopted as the observational parameters. The results showed that SX-115 and CHU-012 induced differentiation of NB4 cells at concentration of 10(-6) mol/L. Comparing the effects of the two retinoids with all trans-retinoic acid (ATRA) at same concentration, there was no significant difference among the three agents. The mechanism of the 2 new retinoids remains possibly the same as ATRA. PMID- 12578680 TI - [Detection of Human Herpesvirus-6 DNA in Peripheral Blood Cells of Patients with Myelogenous Leukemia by Nested PCR] AB - Human herpesvirus-6 (HHV-6) is largly disseminated in human population, which may relate to many diseases. Due to different methods previously used, the results of HHV-6 DNA detections in patients with leukemia are not identical. With a control study, HHV-6 DNA in the blood cells from 38 cases, including 30 cases with acute myeloid leukemia and 8 cases with chronic myelogenous leukemia was detected by nested PCR. The results showed that HHV-6 DNA positive rate (63%) in patients with AML was lower than that in controls, whereas no significant difference between patients with CML (75%) and controls (92%) was shown. The clinical importance of HHV-6 infections in myelogenous leukemia should continue to be investigated later. PMID- 12578681 TI - [Expression of the Cholinesterase-Related Cell Division Controller Gene in Peripheral Blood Cell from Patients with Myelodysplastic Syndrome] AB - To investigate the expression of the cholinesterase-related cell division controller (CHED) gene in the patients with myelodysplastic syndrome (MDS), CHED gene expression was assayed by RT-PCR and its relative expression rate (RER) was determined by the semi-quantitative RT-PCR analysis in peripheral blood mononuclear cells from 21 patients with MDS, 12 normal individuals served as controls. Results showed that RER of CHED in the patients (2.69 +/- 0.76) was significantly higher than that in controls (1.12 +/- 0.51, P < 0.01), the RER out of 85.7% of the patients was higher than the mean value of the controls, in which three patients developed into acute leukemia; the RER out of 61.9% of the patients was higher than the upper limit of the mean value of the controls; three patients whose RER was lower than the mean value of the controls did not developed into leukemia. These findings suggested that the expression of CHED gene in patients with MDS was significantly higher than in controls. PMID- 12578682 TI - [The Feature of Skewing Distribution of TCR Vbeta T Cells from Peripheral Blood and Bone Marrow in Patients with AML-M(5)] AB - The TCR Vbeta 24 subfamily genes were amplified in peripheral blood and bone marrow mononuclear cells from 5 cases with acute monocytic leukemia (AML-M(5)) using RT-PCR, to observe the distribution of TCR Vbeta subfamilies. The results indicated that 1 - 19 Vbeta subfamily T cells could be identified in different samples from AML-M(5) cases. The variation of distribution of TCR Vbeta subfamily T cells could be found in different individual samples. The results provided the feature of cell immune function change in skewed distribution of TCR Vbeta subfamily T cells from peripheral blood and bone marrow of patients with AML M(5). PMID- 12578683 TI - [The Experimental Study on the TCR Idiotypic DNA Vaccine to Induce Antitumor Immune Response to Lymphocytic Malignancy] AB - This study was undertaken to investigate the anti-lymphocytic malignacy immunologic effects induced by TCR idiotypic DNA vaccine on BALB/c mice. CEM lymphoma cell line and BALB/c mice were used as models. The rearrangement gene fragment coding TCR Vbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into the eukaryocytic expression vector pcDNA3 to be used as DNA vaccine. The experimental animals were immunized by intramuscular injection with DNA vaccine. The specific anti-idiotypic antibody was detected by indirect immunofluorescence assay. The specific anti-idiotypic cellular immunity was detected by CTL activity assay using MTT method. The results showed that specific anti-idiotypic antibody in the immunized mice sera could be found since four weeks after immunization and came to the peak of titer on the sixth week. Using IL-6 as immunological adjuvant could significantly increase the antibody titers. It was concluded that the TCR idiotypic DNA vaccine could induce effectively the specific anti-lymphoma idiotypic antibody in BALB/c mice. Using IL-6 as immunological adjuvant could significantly increase the antibody titers induced by idiotipic DNA vaccine. PMID- 12578684 TI - [The Comparison between the Killing Effects of Two Anti-T Immunotoxins on Target Cells] AB - The key to killing target cells by immunotoxin depends on the specific recognition of antibody to target cell and the cytotoxic effect of toxin. The comparative study of the killing effects of two anti-T immunotoxins, CD5:Ricin and CD5:rRA, on target cells was performed. The elimination rate of immunotoxins was analysed by flow cytometry and MLR. The effect of immunotoxins on the proliferation of hematopoiesis was evaluted by CFU-GM. The results showed that (1) CD5(+) T cells were eliminated and CD25(+) CD3(+) activated T cells were concentration-dependently inhibited by the two immunotoxins in the range of 10( 9) - 10(-11) mol/L; (2) both immunotoxins significantly inhibited the mixed lymphocyte reaction, and the inhibiting effect of CD5:rRA to T cell proliferation was markedly lower than that of CD5:Ricin in the range of 10(-10) - 10(-11) mol/L; (3) the combination of CD5:rRA with 10 mmol/L NH(4)Cl increased the T cell elimination rate; and (4) the two immunotoxins and the combination of NH(4)Cl and CD5:rRA did not suppressed proliferation of granulocyte-macrophage progenitors in the range of concentrations with killing effect. It was concluded that T cell and activated T cell could be eliminated effectively by immunotoxins, the proliferation of granulocyte-macrophage progenitor was not inhibited significantly. PMID- 12578685 TI - [Expression of the Wilms' Tumor Gene WT1 and Detection of Minimal Residual Disease in Acute Leukemia] AB - The WT1 gene encoding a zinc finger DNA-binding protein was identified as a tumor suppressor gene being responsible for Wilms' tumor. Recently, aberrant expression of WT1 gene and an inverse correlation between its expression levels and prognosis have been demonstrated in acute myeloid leukemia (AML), suggesting it is a novel tumor marker for leukemic blast cells. To explore whether the WT1 may be used as a marker for detection of minimal residual disease (MRD) in acute leukemia, we examined the sensitivity of the nested reverse transcriptase polymerase chain reaction (RT-PCR) by using WT1 gene primers in comparison with tumor-specific marker genes, such as PML/RARalpha gene in NB4 cells or bcr-abl gene in K562 cells. In all samples, the integrity of RNA was confirmed by amplification of the c-abl gene as an internal control. The limits in amount of leukemic cells detected by two-step RT-PCR with primers for WT1 or tumor specific fusion gene were 10(-4) and 10(-5) in NB4 cells and 10(-3) to 10(-4) and higher than 10(-6) for K562 cells, respectively. None was WT1 positive in peripheral blood mononuclear cells (MNC) from 29 blood donors, while bone marrow MNCs from eight of 21 cases (38.1%) of nonmalignant patient WT1 gene expression were found. Our results suggested that monitoring of WT1 expression makes it possible to rapidly assess the effectiveness of treatment and follow up MRD in AML cases regardless of the presence or absence of tumor-specific markers. PMID- 12578686 TI - [Cytogenetic Diagnosis of Fanconi's Anemia-Distinguishing Fanconi's Anemia from Aplastic Anemia] AB - Fanconi's anemia (FA) is an autosomal recessive disease featuring a great diversity of clinical symptoms, including congenital malformation, growth retardation and bone marrow failure. Cells obtained from FA patients show a specific hypersensitivity to crosslinking agents such as mitomycin C (MMC). In this study, MMC-induced chromosome breakage tests have been done on 27 healthy controls and 51 patients with bone marrow failure [including 48 patients with aplastic anemia (AA) and 3 patients with FA before cytogenetic analysis]. The results showed that: (1) Diagnosis of 4 FA cases was confirmed, and one of them was the correction of clinical misdiagnosis; bone marrow failure combined with congenital malformation was observed in a few of non-FA aplastic anemia patients, while 1 FA patient lacked congenital abnormality and underdiagnosed before cytogenetic analysis. The data confirmed that misdiagnosis or underdiagnosis of FA could be caused without cytogenetic study. (2) Spontaneous chromosome breakages observed in FA patients were the same as those in AA patients and healthy controls. MMC-induced chromosome breakages observed in FA patient cells were much higher than those in AA patients and healthy controls, especially, metaphases containing more than 5 breakages were easily found in FA lymphocytes treated with 50 ng MMC. (3) Mosaic was found in one of the 4 FA patients. MMC induced chromosome breakage test at different MMC concentrations could help to dignosis of FA mosaic patient. PMID- 12578687 TI - [The Gene Chip Technology and the Prospects of Its Application in Hematology and Oncology] AB - The basic principle, technological procedure and types of gene chips were introduced in the article. The probe choice in practical application and processing and hybridization of detected samples were described. Some researchers have used gene chips in hematology, oncology and cellular differentiation. Gene chips can be used to detect the expression of oncogenes, tumor suppression genes, and cell differentiation- and apoptosis-related genes. It can also help us to further study the association between the polymorphism of human genes and disease susceptibility. The aspects of development and problems in gene chips study were discussed. PMID- 12578688 TI - [Progress in Study on MHC Structure and Function] AB - The MHC molecules were first discovered as major histocompatibility antigens. When their essential biological function as antigen-binding molecules and informers for T cells was unveiled, an explanation of why MHC also acts as major histocompatibility antigens was provided. When the antigen-binding cleft of MHC molecules was firstly clarified more than 10 years ago, the study of MHC structure and function has stepped into a prosperous new era, and many achievements have in this area have been achieved. This paper summarized the exciting progress in study on MHC structure and function in recent years. PMID- 12578689 TI - [T-Cell Regeneration after Allogeneic Hematopoietic Stem Cell Transplantation and Its Significance] AB - Allogeneic hematopoietic stem cell transplantation is curative therapy for many malignant hematologic and genetic disorders. The pathways by which T cell are regenerated in vivo are of central importance for host immune competence after the transplantation. Thymic-dependent and thymic-independent pathways of T-cell regeneration are primarily determined by thymus function, and that thymic independent pathways are generally inadequate for restoration of host immunocompetence. T cell regeneration of adult recipients is largely through thymic-independent pathways due to reduced thymic regeneration capacity, resulting in quantitative defficiencies in T-cell number and severe restriction in the diversity of the regenerated T-cell receptor (TCR) repertoire. New strategies to enhance thymic function in man after BMT would hold great therapeutic potential. Activation of donor T cells is the cause of GVHD, the induction of anergy can inactivate specific sets of alloreactive T cells in the donor stem cells, which can prevent GVHD without inhibiting the entire T-cell repertoire. PMID- 12578690 TI - [alpha-Galactosidase and Its Application in Blood Transfusion] AB - alpha-galactosidase (alpha-Gal) belongs to exoglycosidases, which distributed widely in the nature. alpha-Gal can be classified into two types: alpha-Gal A and alpha-Gal B. In vivo, alpha-Gal A is responsible for the cleavage of terminal alpha-galactosidic linkages in glycoconjungates; alpha-Gal B is an alpha-N acetylgalactosaminidase in fact because it hydrolyses alpha-N-acetamidodeoxy-beta D-galactosidic residues from the terminals of a variety of complex carbohydrates and glycoconjugates. Now progress is being made of these two isoenzymes, including studies on blood group conversion, xenotransplantation of organ and pathophysiology of lysosomal storage diseases. There will be significant improvement in the future clinical application of alpha-Gal. PMID- 12578691 TI - [Expression of TFAR19 in Apoptotic Processes of Jurkat Cells Induced with Various Methods] AB - A new apoptosis-related gene TFAR19 was recently cloned. A primary functional analysis indicates its role in the apoptotic process of TF-1 cells. To clarify TFAR19 was involved in which apoptotic pathway, the changes in TFAR19 expression were observed during the apoptotic processes of Jurkat cells induced with various methods: serum deprivation, VP-16 treatment and Fas McAb activation. Then TFAR19 expression in Jurkat cells was detected with flow cytometry and Western blot, and TFAR19 mRNA with RT-PCR. Our results showed that expression of TFAR19 in Jurkat cells was increased at 12 hours after serum deprivation, 2 hours after VP-16 treatment and 2 hours after Fas McAb activation, respectively. These observations suggested that TFAR19 was involved in the apoptotic process of Jurkat cells induced with serum deprivation, DNA destruction and death receptor activation. It might take effect in the early apoptotic process. TFAR19 is a participant of the "final common pathway" in the apoptotic pathway. PMID- 12578692 TI - [beta1 Integrin Dysfunction in Adult Chronic Myeloid Leukemia Bone Marrow Cells] AB - According to our previous experiments, Ph(+) chronic myeloid leukemia (CML) cell line K562 cells have defects in beta 1 integrin activation. In order to search the same regularity in Ph(+) CML bone marrow cells, bone marrow mononuclear cells (BMMNC) from 12 cases of Ph(+) CML and 10 cases of normal individuals were studied. Their expression rate of 9EG7 epitope on beta1 integrin post treatment by 8A2 or GM- or G-CSF and cell adhesion ability with soluble fibronectin (FN) were evaluated by flow cytometry; in addition, the effects of CGP57148B, a highly specific ABL tyrosine kinase inhibitor, were observed. Our results showed that 9EG7 expression rate and FN binding rate were very low in all the inactivated cells. The parameter increased markedly post 8A2 activation in both NBMMNCs and CMLBMMNCs, but the degree of increase in CMLBMMNCs was significantly lower than that in NBMMNCs; GM-CSF or G-CSF could significantly increase the parameters in NBMMNCs while had no effects on that in CMLBMMNCs. CGP57148B could increase the beta1 integrin activation potential of CMLBMMNCs but had no effects on that of NBMMNCs. The results indicate that decreased activation potential of beta1 integrin in CMLBMMNCs is the major cause of adhesion defects of Ph(+) CML cells; beta1 integrin functional insufficiency in CMLBMMNCs could not be directly reversed by ABL tyrosine kinase inhibitor CGP57148B. PMID- 12578693 TI - [Constructing a Competitor of BCR-ABL cDNA by PCR Site-Directed Mutagenesis] AB - A competitive mimic of the cDNA of the BCR-ABL fusion gene was constructed, and its feasibility was testified by capillary electropheresis (CE). The 4 bp-shorter mimic was obtained by PCR amplification using a newly synthesized downstream primer analogous to the former one. Mimics of both types of BCR-ABL cDNA were achieved and the validity was verified with restriction endonuclease. And the products of the coamplification PCR could be easily separated by capillary electrophorisis. The mimic can be used to quantitative detection of BCR-ABL gene through competitive RT-PCR in chronic myeloid leukemia. PMID- 12578694 TI - Mesenchymal Stem Cells Derived from Human Bone Marrow Support Hematopoiesis in Vitro. AB - Bone marrow mesenchymal stem cells (MSCs) are multipotential progenitors of connective tissues and bone marrow stroma as well, which implies the modulatory function of MSCs in hematopoiesis. To clarify the contributions of MSCs to hematopoiesis, the methods for isolation and expansion of MSCs were established and long-term bone marrow cultures were performed using irradiated MSCs as the feeder layer. The results here showed that CD34(+) cells from cord blood formed hematopoietic foci adherent to the monolayer. Furthermore, colony-forming cells remained in the coculture of 5 weeks, indicating the maintenance of long-term culture-initiating cells (LTC-IC). Flow cytometry analysis showed that about 1% of the hematopoietic cells in the culture were positive for CD34 and around 15% were CD41a-positive. It is clear that MSCs maintain LTC-IC in vitro and promote differentiation of hematopoietic progenitors especially into megakaryocytic lineage. The preliminary results here demonstrate that MSCs residing in the bone marrow might be a crucial cellular component in the hematopoietic microenvironment. PMID- 12578695 TI - [Synthesis, Cloning and Expression of a Multiple Epitope Antigen of BCR-ABL Fusion Gene] AB - Chronic myeloid leukemia (CML) appears an ideal and exciting immunological target. Novel and rational immunotherapy may therefore play an important adjuvant role in the treatment of CML patients. Peptides derived from the BCR-ABL fusion region have been shown to be immunogenic and are able to stimulate the production of BCR-ABL-specific T cell lines and clones. In this study, A 280 bp multiple epitope region of BCR-ABL fusion antigen was designed and synthesized. This region contains three BCR-ABL antigen epitopes which can bind to HLA-A2, HLA-A3 and HLA-DR11 molecules, respectively, and epitopes of cholera toxin B (CTB) and tetanus toxoid (TT) which are able to elicit vigorous T cell responses. The fusion antigen gene has highly been expressed in E. coli and the purified fusion protein reserved satisfied activity and antigenicity. The results of this investigation provided a basis for further research on the developing specific T cell immunotherapy of CML. PMID- 12578696 TI - [Expression of Bcl-2 and P53 Gene Proteins on Leukemia Cells and Its Correlation with Chemotherapeutic Efficacy] AB - To explore Bcl-2 and P53 gene proteins expression on human leukemia cells and their relationship with chemotherapeutic efficacy, Bcl-2 and P53 gene proteins expression was assayed by ABC immunohistochemical staining. Results showed that the expression rates of Bcl-2 and P53 gene proteins were 67% and 41% respectively in leukemia cells from 52 patients. While there was no difference of Bcl-2 protein level in ALL and ANLL, the P53 protein level was higher in ANLL than that in ALL (P < 0.05). When CML patients got into the blast crisis phase, the level of Bcl-2 and P53 proteins became very high. Compare with previously untreated AL, relapse/refractory AL patients had higher Bcl-2 and P53 protein level, lower marrow complete remission, and was easy to relapse. The expression of Bcl-2 and P53 protein could be used as new predictors of chemotherapeutic efficacy and prognosis in patients with leukemia. The high protein expression of Bcl-2 and P53 demonstrated that CML was conversion to blast crisis phase. PMID- 12578697 TI - [Development of Acute Promyelocytic Leukemia in PML-RARalpha Transgenic Mice] AB - To investigate the leukemogenic potential of PML-RARalpha fusion protein in vivo, hCG-PML-RARalpha transgene was constructed using molecular cloning technique and hCG-PML-RARalpha transgenic mice were generated. The genotype and phenotype of hCG-PML-RARalpha transgenic mice were analyzed by PCR, RT-PCR, morphology of peripheral blood and bone marrow cells, and pathological examination of spleen, liver and bone marrow. As a result, acute promyelocytic leukemia was developed in 3 hCG-PML-RARalpha transgenic mice in 1 - 5 months. The results demonstrated that PML-RARalpha fusion protein plays a crucial role in leukemogenesis. PMID- 12578698 TI - [Cotransduction of Human mdr-1 and Dihydrofolate Reductase Genes into Murine Hematopoietic Cells] AB - Transfer of drug resistance genes into hematopoietic cells is an attractive approach to protect hematopoietic system from the toxic effects by chemotherapeutic agents in cancer patients. In this study, transduction of mdr-1 in combination with dihydrofolate reductase (dhfr) gene was performed, and the expression of exogenous genes and chemoprotection capacity in mouse bone marrow cells were observed. The results showed that approximately 15% of bone marrow cells transfected with the retroviral vector expressed mdr-1 as assayed by flow cytometry. Gene transfer resulted in about 0.9 - 13 fold and 0.5 - 2.6 fold increase in the resistance of CFU-GM to taxol and methotrexate in vitro, respectively (P < 0.05). Moreover, seven months after transplantation to syngeneic mice with mdr-1 and dhfr-transfected bone marrow cells, peripheral blood cells in recipients were still positive for gp170 as evaluated by FACS as well as for mdr-1 and dhfr by PCR amplification. These results indicate that hematopoietic progenitors can be transfected by retrovirus containing mdr-1 and dhfr genes, and that functional drug resistance accompanies their expressions. Furthermore, genetic chimerism might exist in hematopoietic stem cells. In conclusion, transfer and expression of mdr-1 and dhfr genes in bone marrow cells might be applicable in gene therapy research in cancer patients. PMID- 12578699 TI - [p16 (CDKN2) Gene Transfer Mediated by Retrovirus Vector Inhibits Proliferation of Leukemia Cell Line K562] AB - The aim of this work was to test the effect of p16 on the proliferation of leukemic cells and its potential in gene therapy for leukemia. The full-length p16 cDNA was transfered by recombinant retrovirus vector into leukemia cell line K562, which is homozygous p16 deletion and retains functional retinoblastoma (RB) protein. The cell proliferation was tested in liquid and in soft agar culture after transduction of p16 retrovirus. The results showed a strong inhibition of cell proliferation. Phosphorylation of RB protein was also inhibited. The findings demonstrated that p16 (MTS/CDKN2) inactivation is a significant factor in the genesis and progression of leukemia and p16 could be a candidate gene for gene therapy in leukemia. PMID- 12578700 TI - [Effect of Tumor Vaccine of EL-4 Cell Transduced with IL-12 Gene in C57BL/6 Mice] AB - In order to study the vaccine potency of gene-modified tumor cells, IL-12 express vector was constructed by using retrovirus. The vector was transfected into EL-4 thymic lymphoma cells and the effect of transfectant on anti-tumor immunity was investigated. The tumorigenicity of EL-4/IL-12 transfectant in syngeneic C57BL/6 mice was decreased significantly after implanted with EL-4/IL-12 transfectant compared with EL-4/Wt or EL-4/Neo groups (P < 0.001). The systemic protective immunity was induced against the challenge with EL-4/Wt (in 8/10 mice) after the rejection of EL-4/IL-12 in vivo experiment, a stronger CTL activity against EL 4/Wt cells and a weak killer activity against syngeneic Lewis lung carcinoma (LLC) cells were obtained in (51)Cr release assay. In vivo depletion analysis suggested that the decreased tumorigenicity mainly depended on CD4(+), CD8(+) and NK cells. Therapeutic vaccines with EL-4/IL-12 cells could retard the growth of established EL-4/Wt tumors significantly compared with those of EL-4/Neo (P < 0.005). These studies suggested that immunotherapy and gene therapy using IL-12 is effective in hematopoietic malignancy and IL-12 has prospects of application in human cancer treatment in the near future. PMID- 12578701 TI - [Therapeutic Effects on Chemotherapy-Induced Granulocytopenia in Hematologic Malignacies, a Comparison Between Lishengsu and Filgrastim] AB - A multi-center, open controlled study was performed to observe the effects of Lishengsu, a rhG-CSF preparation manufactured by Beijing Shuanglu Biopharmaceutical Co., Ltd. on chemotherapy-induced granulocytopenia in patients with hematological malignancies. The results showed the total response rates were similar between Lishengsu- and Filgrastim-treated groups (98.8% vs 100%), recovery of peripheral ANC and WBC was also similar between the two groups, with the peaks appeared both at the 12th day after rhG-CSF treatment. No differences were observed in the duration for ANC to recover to normal level between Lishengsu- and Filgrastim-treated groups (7 and 8 days respectively). Our results indicated Lishengsu is as effective as Filgrastim in accelerating the recovery of chemotherapy-induced granulocytopenia. PMID- 12578702 TI - [Curative Effect of Lishengsu on Leukopenia and Infection after Chemotherapy in Tumor Patients] AB - To examine the effect of Lishengsu (rhG-CSF) on the recovery of leukopenia and control of infections after chemotherapy, twenty tumor patients were observed. When blood WBC decreased to less than 2.5 x 10(9)/L and ANC less than 1.5 x 10(9)/L after chemotherapy, Lishengsu was administered in a dosage of 150 micro g/day for 5 days or until WBC and ANC recovered to normal level. The changes of hemogram, body temprature, the signs of oral mucous membrane and lung were examined. The recovery of WBC and ANC in the Lishengsu group was much faster than that in the control group. The difference between the two groups was statistically significant (P < 0.01). The duration of fever was shorter and the healing of oral ulcers was faster in Lishengsu group than that in the control group (P < 0.05). The results indicated that Lishengsu in a low dosage and a short administration period could enhance the recovery of WBC and ANC efficiently, shorten the duration of fever, quicken the recovery process of oral ulcers and reduce the needs for supporting treatment in tumor patients after chemotherapy. PMID- 12578703 TI - [Therapeutic Effect of rhIL-11 on Thrombocytopenia Induced by Carboplatin in Rhesus Monkeys] AB - The effectiveness of rhIL-11 on thrombocytopenia induced by carboplatin in rhesus monkeys was investigated. Thrombocytopenia was induced in monkeys by i.v. administration of carboplatin at a dose of 15 mg/kg(-1)/d(-1) for three consecutive days. rhIL-11 (50 or 100 micro g/kg(-1)/d(-1)) or Neumega (100 micro g/kg(-1)/d(-1)) were administered s.c. for 14 days beginning one day following the final dose of carboplatin. The results showed that rhIL-11 significantly improved mean platelet nadirs and shortened the mean duration of platelet counts less than 50% of pre-treatment values. Administration of rhIL-11 also resulted in moderate increase of the reticulated platelet, leukocyte and reticulocyte counts in peripheral blood and megakaryocytic and erythroid progenitors in bone marrow. rhIL-11 did not enhance ADP-induced platelet aggregation. These results indicate that rhIL-11 has a potent thrombopoietic effect in vivo and could be an important agent to reduce the severity and duration of thrombocytopenia following chemotherapy. PMID- 12578704 TI - [Recovery of Early and Mature T Lymphocyte Subsets after Allogeneic Hematopoietic Stem Cell Transplantation] AB - To study the reconstituion of the T-cell populations after allogeneic hematopoietic stem cell transplantation, the peripheral blood samples obtained at different time points post-transplantation in 21 patients were analyzed using CD5 PE and CD3-FITC with flow cytometry. During the first 3 months after transplantation, CD3(-) CD5(+) T cell subsets, representing early thymocytes, remained below normal level, whereas CD3(+) CD5(+) T cell subsets, representing mature T cells, regenerated to normal level. In 9 patients who developed acute graft-versus-host disease (GVHD), the percentage of CD3(+) CD5(+) T cell subsets was significantly higher than that in patients who did not develop acute GVHD (P < 0.05). These results demonstrated that the early recovery of T cells is mainly due to the expansion of mature T cell populations, and the over-expansion of mature T (CD3(+) CD5(+)) cells is responsible for the development of acute GVHD. PMID- 12578705 TI - [Giant Tumor Cells in the Bone Marrow and Their Significance] AB - Giant tumor cells and their varieties in the bone marrow were found in 7 patients with abnomal hematopoiesis phenomena. These cells were artificially devided into 5 kinds according to the difference of their morphology. Most of these cells were corresponding to lymphoid-monocytoid-macrophagocytoid cells with Wright's staining, cytochemical stainings, immunocytochemical stainings, flow cytometry examination, electron microscopy and pathologic study. The bone marrows were hypercellular and marked dysplastic hematopoiesis phenomena. Two of the 7 cases were diagnosed as malignant lymphoma with bone marrow biopsy. All cases characteristically showed no lymph node enlargement or hepatosplenomegaly or any local tumor mass. As to the prognosis of these cases, two patients died with survival time of 8 and 17 months, respectively, one was on critical condition at course of 10 months, and the other 4 cases were in comparatively stable condition with courses of 2.5 to 24 months. These patients seem to be a group of rare malignant lymphoid-monocytoid-macrophagocytoid proliferative diseases. PMID- 12578706 TI - [Study on the Polymorphism of Intron 22 (CA)n Repeat within FVIII Gene in Dai, Yi and Han Populations of Yunnan Province] AB - This study was designed to investigate the polymorphism of intron 22 (CA)n repeat within FVIII gene in Dai, Yi and Han populations of Yunnan province. PCR, DNA sequencing technique and PAGE were used in this study. The results showed that three different alleles corresponding to 24, 25 and 26 dinucleotides were found at this locus and the allele frequency ranged from 1.20% to 57.7% in Dai and from 16.43% to 63.00% in Yi populations. In Han population, five different alleles with 24, 25, 26, 27 and 28 (CA)s were found, the allele frequency ranged from 8.97% to 32.00%. Heterozygotes of intron 22 (CA) repeat within FVIII gene were not found in the three populations. In conclusion, it was obvious that the locus cannot be acted as DNA genetic marker of FVIII gene in the three populations in Yunnan. PMID- 12578707 TI - [Origin and Regulation of Hematopoietic Stem Cells During Embryonic Ontogeny] AB - Stem cells of various tissues including hematopoietic tissue in the body are derived from embryonic stem cells (ESC). There exists intricated gene regulation during ESC development and its differentiation into hematopoietic stem cells (HSC). In embryo hematopoiesis development, there are two kinds of hematopoietic types, primitive hematopoiesis and definitive hematopoiesis. The theory of the yolk sac of primitive hematopoiesis is well accepted, while the initial site of definitive hematopoiesis still exists controversy. In present opinion, there are at least two independent sites associated with definitive hematopoiesis, those are, yolk sac and para-aortic splanchnopleura (PAS)/aorta-gonad mesonephros (AGM). Study on the hematopoiesis and its regulation during embryonic ontogeny will benefit not only to the dicovery of the mechanism of some blood disorders, but as well to gene therapy and HSC engineering. PMID- 12578708 TI - [Advance of Research on Treatment of Donor Lymphocyte Infusions in Relapse of Hematologic Malignancies after Allogeneic Bone Marrow Transplantation] AB - The applications of allogeneic bone marrow transplantation (allo-BMT) are limited by the high risk of relapse with primary malignancy. The newly adaptive immunotherapy with donor lymphocyte infusions (DLI) can induce an effective graft versus-leukemia (GVL) reaction leading to molecular remission and prolonged disease free survival in the majority of patients with relapse of CML and some other hematologic malignancies. Little is known about the mechanisms and the kinetics of GVL, however, increasing evidences show that in the case of existence of chimerism the mature donor T lymphocytes are activated by the specific leukocyte artigen presented in the surface of host antigen presenting cells (APC), resulting in exclusively elimination of leukocyte. Undesired side-effects are the development of graft-versus-host disease (GVHD) and the occurrence of pancytopenia in some patients. Infusions with selective T lymphocytes and G-CSF mobilized peripheral blood stem cells (PBSC) are the promising treatment of DLI for more patients. PMID- 12578709 TI - [Challenges to Hematopoietic Stem Cell Research Today] AB - More works documented recently indicated that human CD34(-) cells exist and are likely to be the precursors of the CD34(+) cells. Anyhow, the CD34(+) enriched populations have already been proved to show long-term reconstitution of hematopoiesis in animals and patients worldwidely. It still remains uncertain whether cells lack of CD34 and Lineage markers are the very best stem cells or maybe the residual embryonic stem cells keeping quiescent in the adult tissues are capable of transfer into hematopoietic stem cells when activated. Since just a negative selection technique is used to collect the CD34(-) cells everywhere for the time being, no final conclusion is convincing about the characterization of CD34(-) cells till a highly purified cell population of CD34(-)/Lin(-) is available. Clinical analysis shows that the most critical factor predicting the stem cell engraftment is the number of the cells infused. The number of nucleated cells in umbilical cord blood to be infused and required to obtain a successful engraftment is superior to 3.7 x 10(7)/kg. However, large dose of T-cell-depleted and purified CD34(+) cells as more than 5 x 10(6)/kg or even a 'megadose' of CD34(+) cells of 10(7)/kg is recommended for allotransplant of mobilized peripheral blood to achieve a high rate of successful engraftment. The delayed engraftment and the relapse of malignancy after cord blood transplantation are major problems. However, CBT is still the best choice of stem cell transplant for the baby patients with non-malignancies. At present, the HLA typing for class I antigens is still achieved with serology in most laboratories. As HLA typing is increasingly defined to higher degree of resolution by DNA probes, it is recommended to check with genotyping when there is a 'match' by the serological phenotyping especially for the unrelated donor/recipient couples. Improvements in DNA-based methods for the detection of numerous HLA alleles have provided the opportunity to investigate the relationship between HLA disparity and transplant complications. About 80% (or even more) of patients in China who might benefit from stem cell transplantation still fail to find suitable donors. It is worthy to adopt the unrelated donors matched or mismatched for those high-risk acute leukemia patients who do not have related matched donors but urgently need transplant. The advanced experience of unrelated mismatched transplant will no doubt be certain to carry weight and be disseminated in China. A great leap forward of the clinical practice and biological study on hematopoietic stem/progenitor cells is expected in the new century to accept the challenge from the world outside China. PMID- 12578710 TI - [The Immunological Study of Megakaryocytes] AB - The immunological role of megakaryocytes is not well known. This project studies the involvement of megakaryocytes on immuno-inflammatory processes and the possible mechanism via the adhesion molecule CD36 and the synthesis of relevant cytokines. The expression of adhesion protein CD36 on human platelets, megakaryocytes and megakaryocytic cell lines (Meg-01, Dami, CHRF-288-11 and M 07e) was analyzed by using flow cytometry, ELISA and immunocytochemical methods. The expression of interleukin-1 (IL-1) to interleukin-10 (IL-10), TNF-alpha, TNF gamma and IFN-gamma on the four megakaryocytic cell lines was also determined by RT-PCR. The effect of IL-1beta, IL-3, IL-6 and TPO on murine megakaryocyte colony formation (CFU-MK) was studied by using a plasma clot culture system. The CFU-MK was confirmed by acetylcholine esterase staining. The results showed that: (1) CD36 was expressed on platelets, megakaryocytes and the four megakaryocytic cell lines, the relative expression level is as follows: platelets > megakaryocytes > Meg-01 > Dami > CHRF-288-11 > M-07e, suggesting that the level of CD36 expression correlates with the degree of maturity of megakaryocytic differentiation; (2) inflammatory cytokines TNF-alpha, IL-1beta, IL-3 and IL-6 were detected in all the four megakaryocytic cell lines, suggesting that different stages of megakaryocytes can be as a source of inflammatory cytokines; and (3) IL-1beta, IL 3 and IL-6, as well as TPO, play a stimulating effect on CFU-MK formation, suggesting that there is an "autocrine" effect on megakaryocytopoiesis. The data obtained suggest that megakaryocytes may involve in immuno-inflammatory processes via the synthesis of platelet adhesion molecules and inflammatory cytokines. PMID- 12578711 TI - [Control of Thrombopoietin Expression by Doxycycline in CHO Cells] AB - To control thrombopoietin (TPO) expression by doxycycline in CHO cells, Tet-On gene expression system was used. Recombinant plasmid pTRE-TPO was successfully constructed. pTRE-TPO and pTK-Hyg were cotransfected into CHO-Tet-On cells. Cells resistant to hygromycin were cloned, high expression and low background clone was selected, and named as CHO-Tet-On-TPO. There was no significant TPO expression in the absence of doxycycline, the TPO level in the cell culture supernatant was 0.1 micro g/L. After exposure to 2 mg/L doxycycline, TPO expression was greatly increased, the TPO level in the cell culture supernatant reached 10.8 micro g/L. Tet-On gene expression system is a ready access to the tight-regulated and high level gene expression system. It is likely to provide a safe and regulatable way for TPO gene therapy. PMID- 12578712 TI - [Construction of Expression Vector for Human CD40-Ig Fusion Protein and Its Expression in COS-7 Cells] AB - CD40/CD40L, besides B7/CD28, is an alternative important costimulation signal transduction pathway. It plays a pivotal role in T cell activation. Moreover, it may play a critical role at many levels of sensitization and effector phases of allograft rejection. In order to get the fusion protein of human CD40 extracelluar region and IgG 1 Fc fragment, and investigate the potential role of blocking CD40/CD40L costimulation pathway in immunotherapy, total RNA was extracted from human lymphoma cell line Daudi, and CD40 gene extracelluar region was amplified by RT-PCR. The PCR products were inserted into pGEM T Easy vector, and the cloning vector pGE40 was obtained. The DNA sequence was analyzed by automatic DNA sequencer. After sequencing, the transient expressing vector was constructed by inserting correct fragment into pIG vector, which contains the genomic human IgG1 Fc (hinge, CH2 and CH3) gene. Hence the recombinant fusion expression vector was constructed successfully, and named after pIG/40 Ig. Then, COS-7 cells were transfected through DEAE-Dextran/chloroquine method. The CD40-Ig fusion protein expressed in COS-7 cell culture supernatant was identified by sandwich ELISA and Western blot. Result showed that the CD40-Ig fusion protein can be detected by sandwich ELISA in the cell culture supernatant. Western blot analysis also showed that it could react with McAbs of mouse anti-human CD40 G28 5 and mouse anti-human Ig gamma chain. There is only one obvious band at the position of relative molecular weight 50 kD, and it is equivalent to the expected value. Above all, the recombinant fusion expression vector pIG/40 Ig was constructed, and CD40-Ig fusion protein gene was expressed in COS-7 cells successfully. It could be laid a foundation to investigate the potential role of CD40/CD40L pathway as the target of GVHD prevention and therapy. PMID- 12578713 TI - Bone Marrow Endothelial Cell-Derived Factors Inhibit the Growth of Marrow CFU-F. AB - The present study investigated the effects of the serum-free conditioned media of the bone marrow endothelial cells on CFU-F for potential mechanisms upon which hematopoiesis may be regulated by them within the bone marrow microenvironment. After obtaining the serum-free conditioned media of human and murine purified bone marrow endothelial cells (hBMEC-CM and mBMEC-CM) in vitro, MW > 10 kD, 3 - 10 kD and < 3 kD components were sifted out from these media by means of serial ultrafiltration. Assays of CFU-F were performed to test the effects of BMEC-CM and their ultrafiltrated components. The results showed that every one of hBMEC CM, mBMEC-CM and their MW < 3 kD components exerted a suppressive effect on the proliferation of corresponding CFU-F but MW > 10 kD and 3 - 10 kD components. The BMEC-CM and MW < 3 kD components decreased the number as well as the size of CFU F. There were the markedly negative concentration-dependent relations between the concentrations of MW < 3 kD component and the numbers of CFU-F. These findings suggested that bone marrow endothelial cells in culture could secrete at least a humoral factor (molecular weight less than 3 kD) which has an inhibitory effect on the growth of CFU-F. PMID- 12578714 TI - [Promotion of Thrombopoiesis by rhIL-11 in Normal and Myelosuppressed Mice] AB - Hematological effects of rhIL-11 on normal and myelosuppressed male BALB/c mice were observed. Mice were subcutaneously injected with rhIL-11 for 7 consecutive days, at the dose of 200 or 400 micro g/kg per day, peripheral blood platelet counts were moderately elevated on 5 days after administration and returned to base level within 4 days after discontinuation of injection. In myelosuppressed mice, treatment with rhIL-11 significantly ameliorated the degree of thrombocytopenia, the recovery of thrombocytopenia was also significantly accelerated at the dose range of 100 - 400 micro g/kg per day, and blood platelet counts reached pre-irradiated level after 13 - 15 days of treatment. The magnitudes of platelet count elevation were similar among groups of 100, 200 and 400 micro g/kg per day, although recovery appeared earlier in group of 400 micro g/kg per day. Significant increases in CFU-Meg were observed both in normal and myelosuppressed mice. Our results suggest that rhIL-11 promotes the increase of peripheral blood platelets both in normal and myelosuppressed mice, and can be used as a potential therapeutic agent for thrombocytopenia induced by chemotherapy. PMID- 12578715 TI - [Therapeutic Effects of rhIL-11 on Carboplatin-Induced Thrombocytopenia in Monkeys] AB - A model of myelosuppression with thrombocytopenia was produced in monkey by i.v. administration of carboplatin to the evaluate effects of rhIL-11 treatment in monkeys. Following myelosuppression, rhIL-11 was subcutaneously injected for 19 consecutive days at the dose of 50 or 100 micro g/kg. In myelosuppressed monkeys treated with rhIL-11, peripheral blood platelet started to drop at the day 8 after the administration of carboplatin, and reaching the nadir between the day 12 - 14, the decrease in blood platelet was less severe compared with untreated monkeys; peripheral blood platelet began to recovery on day 11 - 13 (D14 - D16) after rhIL-11 treatment, and reached or surpassed the baseline value before carboplatin administration after 13 - 15 days rhIL-11 treatment. Blood platelet counts remained high level after discontinuation of rhIL-11 administration and returned to baseline after 4 days. The results demonstrated that rhIL-11 has a significant thrombopoietic activity, it can reduce the severity of thrombocytopenia as well as shorten the duration of thrombocytopenia caused by myeloablastive agents, and is likely to become an effective agent against thrombocytopenia induced by chemotherapy. PMID- 12578716 TI - [Synergistic Induction of Apoptosis by Chemotherapeutic Drugs and Cytokines in Mouse T-Lymphoma Cell Line] AB - The objective of the study is to find out the synergistic effect on apoptosis resulting from the combination of chemotherapeutic drugs and some cytokines. Dexamethasone (DEX), etoposide (VP16), arsenic trioxide (As(2)O(3)) and alltrans retinoic acid (ATRA) were added to the murine T lymphoma cell line RMA as well as to the cells preincubated with IL-2, IL-6 or GM-CSF, respectively. The effect on apoptosis was observed. All four chemotherapeutic drugs inhibited the cell proliferation. DEX, VP16 or As(2)O(3), except ATRA, singly induced apoptosis of RMA cells. The DEX concentration of inducing apoptosis was reduced when it was added together with ATRA. IL-2, IL-6 and GM-CSF did not induce apoptosis when the cytokines were added to RMA separately, however, apoptosis could be induced by combination of IL-2 and IL-6. The cytokines facilitated the apoptotic action of chemotherapeutic drugs, the drug concentration for inducing apoptosis decreased and the time period of starting apoptosis shortened. The results demonstrated that there was synergistic effect of chemotherapeutic drugs and some cytokines for induction of apoptosis. It raised an experimental basis for combination of chemotherapeutic drugs and some cytokines, especially IL-2, in the treatment of malignant lymphoma. PMID- 12578717 TI - [Effect of Methotrexate on Cell Growth of Human Monocyte-like Cell Line U937 and Its Induction of Apoptosis] AB - Acute myeloid leukemia (AML) is considered to be a malignancy that is intrinsically resistant to methotrexate (MTX). As compared to acute lymphoblastic leukemia (ALL) blasts, AML blasts, except those of acute monocytic leukemia (AML M(5)), form fewer amounts of long chain polyglutamate of MTX (MTXPG), when incubated with MTX, thus providing an explanation for their lack of responsiveness to MTX. To explore the novel approach of treatment in patients with AML-M(5), the U937 cell line, which has the monocytic characters, was used. Cell growth inhibition was mearsured by XTT assay after 24 and 48 hours in the continuous presence of various concentrations of MTX ranging from 1 nmol/L to 100 micro mol/L. After 24 hours MTX treatment, the IC(50) value for U937 cells was 0.04 micro mol/L. After 48 hours treatment, the IC(50) was 0.037 micro mol/L and IC(90) was 0.39 micro mol/L. To understand the mechanism of MTX cytotoxicity, the process of cell death was analyzed. A variety of assays, including trypan blue exclusion, flow cytometry, light microscopy (Wright's staining) and DNA fragment electrophoresis, were performed. There were no significant apoptotie changes after shorter exposure of MTX (4 and 6 hours). After 8 hours at various concentrations of MTX treatment ranging from 5 nmol/L to 10 micro mol/L, the percentage of the cells in the pre-G(1) (apoptotic) was 3.2% at 0.1 micro mol/L and it reached a peak of 18.2% at 5.0 micro mol/L. The DNA synthesis in S-phase was inhibited from 41.2% (0.01 micro mol/L) to 19.1% (10 micro mol/L). DNA ladder band, a feature of apoptosis, was observed. The arrest of cell growth and apoptotic properties induced by MTX have lead to its evaluation as a potentially therapeutic agent in the treatment of AML-M(5). PMID- 12578718 TI - [Expension In Vitro of T Cells from Cord Blood CD34(+) Cells Stimulated with SCF and IL-2] AB - The generation of large quantities of novel human T cell clones ex vivo would make a wide range of gene-and immuno-therapies for tumor and AIDS possibly. Although it is well established that T cells are derived from CD34(+) cells, the involvement of thymic fragments from either human or murine fetus makes the in vitro T cell perliferation very cumbersome. In this report, cord blood mononuclear cells were used as accessory cells to support the differentiation of CD34(+) cells into naive T cells stimulated with SCF and IL-2. CD4(+) and CD8(+) T cells, under the cultural conditions, were continuously produced in vitro at least over a period of 3 weeks and their ratios changed gradually. CD4/CD8 double positive T cells and RAG-2 gene were existed, and RAG-2 gene, reponsible for TCR rearrangement, was expressed during the cell proliferation. Our study presents a simple culture system in vitro to acquire large quantities of naive T cell clones. PMID- 12578719 TI - [A Nude Mouse Model for Human Umbilical Cord Blood Transplantation] AB - To evaluate the hematopoietic potentiality and the migration and homing routine of separated as well as cryopreserved umbilical cord blood hematopoietic cells, the BALB/cnu(+) mice were used to establish a murine model. This can prepare for the clinical transplantation and the establishment of a large-scale cord blood bank. The result indicated that the hydroxyethyl starch (HES) sedimentation and DMSO step-by-step cryopreservation procedure resulted in only less losses of hematopoietic progenitor cells and also unharmful to the hematopietic potentiality. We can found evidence for successful transplantation in each mouse which received (1.0 - 2.0) x 10(7) separated or cryopresered hematopoietic cells from cord blood, which lasted for about fifty days. The results demonstrated that (1) HES sedimentation and DMSO cryopreservation procedure can keep the hematopoietic potentiality of cord blood, and so can be used to clinical transplantation or establishment of a cord blood bank; (2) Rich hematopoietic stem cells in human cord blood can cross the xenogenetic barriers and successfully engraft mice; (3) The hematopoietic cells migrated among bone marrow, liver, spleen, lung and kidney in the mice and homed to bone marrow by the end. Cryopreservation may influence the adhesion molecule on the hematopoietic cells and the homing behaviour, but not influence their hematopoietic potentiality. PMID- 12578720 TI - [Effects of Allogeneic Transplantation of T Helper-2 Cells on Graft-versus-Host Disease and Graft-versus-Leukemia Effect] AB - This study was undertaken to explore whether the graft-versus-host-disease could be decreased and graft-versus-leukemia effect be retained by transplantation of allogeneic T helper-2 (Th2) cells. T cells from C57BL/6(H-2b) mice were incubated and polarized with rmIL-4, Con A and ionomycin in vitro, and then, the T cells were mixed with marrow cells and transplanted into recipient BALB/c(H-2d) mice bearing erythroleukemia cells. The occurence of GVHD and GVL effect was observed. The results showed that the mean survival time in the groups of untreated control, cyclophosphamide treatment, marrow and spleen T cell transplantation and marrow and Th2 cell transplantation was 10.6 +/- 1.3, 18.7 +/- 4.2, 22.7 +/- 7.4 and 36.9 +/- 10.8 days, respectively. In untreated control and cycophosphamide treatment groups, all of ten mice died from leukemia. Nine of ten mice died from GVHD in marrow and spleen T cells transplantation group. In marrow and spleen Th2 cell transplantation group, three of ten mice died from GVHD, and GVHD was not occurred in the other seven mice, and there was no any evidence of leukemia in two mice on 50 days after transplantation. It was concluded that tranplantation with polarized Th2 cells could relieve GVHD, and at the same time retain the GVL effect. PMID- 12578721 TI - [Prospects of Research on Bone Marrow Mesenchymal Stem Cells] AB - Besides hematopoietic stem cells, bone marrow also contains another type of stem cells called mesenchymal stem cells (MSCs). With different induced conditions, MSCs have the ability to differentiate into a variety of nonhematopoietic tissue cells, including osteoblasts, chondroblast, adipocytes, myoblasts, astrocytes, and so on. MSCs can be readily obtained from bone marrow by their adhesion to plastic and expansion in culture. Also they can be genetically engineered by transduced target genes. MSCs may be the farget cells for both cell therapy and gene therapy for diseases derived from many different nonhematopoietic tissues. PMID- 12578722 TI - The Role of Transcription Factor GATA-2 in Early Hematopoiesis. AB - Transcription factor GATA-2 is a member of GATA family which binds a common DNA sequence motif [T/A (GATA) A/G] through an evolutionarily conserved zinc finger domain. An essential role for GATA-2 in the development of hematopoietic cells has recently been shown in gene knockout experiments in mice. GATA-2 gene disruption makes reduction in all hematopoietic precursors, whereas enforced expression of GATA-2 blocks normal hematopoiesis. GATA-2 exerts regulation in early embryo and cooperates with other GATA transcription factors in proliferation and differentiation of myeloid, erythroid, megakaryocytic and mast cell lineages. GATA-2 mRNA and protein could be detected in human myelogenous leukemia cell lines and in most patients with primary leukemias. Some data revealed that GATA-2 could transactivate the Cas-Br-E and graffi retroviruses, which can induce myelogenous leukemia in mice. However, the role of GATA-2 in pathogenesis of leukemia is still not well understood. PMID- 12578723 TI - [Study on Gene Defects in Myelodysplastic Syndrome] AB - Myelodysplastic syndrome (MDS) is a clonal disorder of the hematopoietic stem cell. Recent developments in molecular genetics have provided insights into the molecular mechanisms that lead to MDS. This review summarizes the most recent developments in the understanding of the gene defects associated with MDS which mainly include altered expression of oncogenes, the loss of expression of tumor suppressor genes and the repair of DNA. PMID- 12578724 TI - [Advance in Reseach on Human Immunodeficiency Virus Type I Vector] AB - Human immunodeficiency virus type-I (HIV-I), one kind of lentiviruses, was characterized by a complex genome that encodes two regulatory proteins and four accessory proteins in addition to the common gag, pol and env gene products. So far, a few of different types of replication-defective vectors were constructed, the highest viral titer from one of which was above 10(7) TU/ml. Several studies on packaging cell line found that eliminating the four accessory genes would have few effect on transduction ability of vector and split-genome package system could reduce the possibility of producing replication-competent virus. There are two kinds of characters on HIV-I vectors. Firstly, it was highly efficient in transducing CD34(+) human hematopoietic stem/progenitor cells; secondly, repeated injections of the HIV-I vector into animal did not elicit the rejection response. HIV-I vector had an extensive host range. PMID- 12578725 TI - The role of complement and natural antibody in intestinal ischemia-reperfusion injury. AB - Activation of the complement cascade is central to many types of injury. Ischemia reperfusion is an important example of such an event. Using intestinal ischemia reperfusion as a model, we have further elucidated the importance and mechanism of this activation. Of novel importance is the evidence that natural antibody is a trigger for these events via recognition of self-antigen. In this article, we review the role of natural antibody and complement in intestinal ischemia reperfusion injury. It is hoped that this study will ultimately lead to better understanding of these important modulators and their role in this type of injury. PMID- 12578726 TI - Managing failure to antiretroviral drugs in HIV-1-infected patients. AB - Managing failure to antiretroviral therapies implies the addressing of several issues: the clinical stage, the virological and the immunological response to the failing regimen, together with drug history, resistance and exposure. Each of these issues will be discussed with the aim of providing useful data to design an optimal rescue antiretroviral therapy. PMID- 12578727 TI - Immediate allergic reactions to beta-lactams: diagnosis and therapy. AB - Beta-lactams are the antibiotics which most frequently provoke adverse reactions mediated by specific immunological mechanisms. These reactions, classifiable as immediate or non-immediate, can be produced by the four classes of beta-lactams (penicillins, cephalosporins, carbapenems and monobactams) currently available, which share a common beta-lactam ring structure. Immediate reactions occur within the first hour after drug administration and are characterized by urticaria, angioedema, rhinitis, bronchospasm, and anaphylactic shock. Immediate reading skin tests are the quickest and most reliable method for demonstrating the presence of beta-lactam specific IgE antibodies. It is crucial to use in diagnosis the suspected beta-lactams themselves, particularly cephalosporins, in addition to penicillin determinants. Serum specific IgE assays can be used as complementary tests. Negative test results should be interpreted in light of the time elapsed from the last exposure to the responsible beta-lactam. In fact, both in vivo and in vitro test sensitivity is known to decrease over time. In some diagnostic work-ups, patients with a positive history and negative skin and in vitro tests with classic reagents undergo a controlled administration of the suspected beta-lactam. The management of immediate allergic reactions should take into consideration their severity and type. Adrenaline is the drug of choice in the treatment of anaphylactic shock. In addition to adrenaline, corticosteroids and antihistamines should be administered. Histamine H(1) receptor antagonists are the mainstay of the treatment of immediate allergic reactions such as urticaria, rhinitis and conjunctivitis. PMID- 12578728 TI - Atherosclerotic plaque formation and risk factors. AB - Atherosclerosis (AT) is a progressive disease characterized by the accumulation of lipids, fibrous materials, and mineral in the arteries. Although many generalized or systemic risk factors predispose to its development, AT affect various regions of the circulation preferentially and yields distinct clinical manifestations depending on the particular circulatory bed affected. The progression of AT is currently believed to involve the interaction of endothelium, monocytes, and leukocytes, as well as the influences of cytokines, oxidized lipoproteins, hypertension, tobacco use, dyslipidemia, homocystinemia, and genetic determinants. PMID- 12578729 TI - Cryoglobulinemia: a true internistic disease? AB - Cryoglobulinemia is a clinical condition characterised by the presence of circulating globulins that precipitate at a temperature lower than 37 degrees Celsius and re-dissolve with warming. We can distinguish 3 different types of cryoglobulinemia, according to their immunochemical characteristics. Cryoglobulinemia can be associated with infectious, inflammatory or neoplastic disease. Cryoglobulinemia type II can be associated with chronic HCV-hepatitis. Clinically, cryoglobulinemias cause hyperviscosity-related symptoms or lesions by immunocomplex deposition (cryoglobulinemic vasculitis). Many organs and systems can be involved, from the skin to the joints, from the central nervous system to the kidney. Diagnosis requires a careful clinical and physical evaluation and the demonstration of circulating cryoglobulins by cryoprecipitation and immunoelectrophoresis. The therapeutic goals are the treatment of the underlying diseases and the complication and prevention of progression/relapse. It is obvious that this disorder can involve different specialists, but the internist plays a central role: he identifies the disease and the associated condition, he treats the underlying disorder and refers the patient to the specialists for the organ-specific manifestations. PMID- 12578730 TI - Inhibitory effect of retinoic acid on proliferation, maturation and tryptase level in human leukemic mast cells (HMC-1). AB - Mast cells play important role in allergic inflammation by releasing histamine, tryptase and several inflammatory cytokines. Human leukemic mast cells (HMC-1) have been used to study mast cell mediator and their role in inflammatory mechanisms. HMC-1 contain and release several inflammatory mediators, of which the proteolytic enzyme tryptase is most characteristic. Retinoids, including retinoic acid, are naturally occurring and synthetic derivatives of vitamin A. All-trans-retinoic (ATRA) acid had been previously reported to inhibit cell proliferation, differentiation and apoptosis. In the present study, we investigated the effect of ATRA on the proliferation and secretion of tryptase in HMC-1. HMC-1 were treated with ATRA at 10(-4M), 10(-5M) or 10(-6M) for 3, 4 or 5 days in culture. Control HMC-1 were treated with equal amount of culture medium only. ATRA decreased the number of HMC-1 as compared to the control group. The same treatment for 3, 4 or 5 days also decreased intracellular tryptase levels. These results indicate that ATRA significantly inhibits both proliferation and growth as shown by the decreased intracellular tryptase levels in HMC-1. ATRA may be a useful agent in the treatment of mast cell proliferative disorders. PMID- 12578731 TI - Levels of enzymatic antioxidants activities in mononuclear cells and skin reactivity to sodium dodecyl sulphate. AB - Chemical irritants are able to produce several biological modifications of the skin, including the direct or indirect production of cytokines and reactive oxygen species leading to an inflammatory reaction. This report examines the existence of a possible correlation between the skin sensitivity to the irritant sodium dodecyl sulphate (SDS) and the activity of the enzymatic antioxidants. In twenty-three healthy subjects the evaluation of the epidermal and peripheral blood mononuclear cells (PBMCs) activities of Superoxide Dismutase (SOD) and Catalase (Cat) demonstrate a significant correlation (r= 0,85 and p< 0,005 for SOD, and r= 0,87 and p< 0,0001 for Cat). Based on this result, on a further group of normal subjects (n=13) we studied the link between the threshold dose of skin reactivity to SDS and the activities of the enzymatic antioxidants in PBMCs. The degree of skin modification induced by SDS, applied at different concentrations for 24 hrs, was determined by means of Trans Epidermal Water Loss (TEWL), Erythemal Index or by Visual Score (VS). The minimal dose of the irritant capable of inducing skin modifications, was significantly correlated with SOD (r=0,77) and Cat (r=0,81) activities in PBMCs, and the modification of TEWL or EI were inversely correlated with levels of antioxidants in PBMCs (r=-0,62 for SOD and r= 0,66 for Cat). Our results indicate that the skin reactivity to irritants can be modulated by the levels of antioxidants, and suggest a possible therapeutical approach in preventing irritant contact dermatitis. PMID- 12578732 TI - Purification and characterisation of two GST's forms from Rhizobium leguminosarum with a high affinity to herbicides. AB - Cytosolic glutathione transferases are a family of multifunctional proteins that catalyse the conjugation of GSH to a large variety of endogenous and exogenous compounds. These enzymes have been widely studied in mammals and, to a lesser extent, in plants. In plants, GSTs can detoxify herbicides; they are also induced by pathogenic infection and are likely to be involved in defence responses. GSTs are found in pathogenic and not pathogenic prokaryotes but the functional role played by these enzymes in the cell still remains to be clarified. Here we report the purification and characterisation of two GST forms from Rhizobium leguminosarum that play a very important role in agriculture by inducing nitrogen fixing nodules on the roots of legumes. These bacterial GSTs from R. leguminosarum have immunological characteristics that are different among them and they are characterised both by a high affinity to herbicides. PMID- 12578734 TI - The role of substance P in cerebral ischemia. AB - Tackykinins are involved in the inflammatory process of a large number of diseases. The role of the tachykinins in ischemic brain injury was evaluated by the serum levels of Substance P (SP), one of the most known tachykinins and detected by a competitive enzyme immunoassay. The study was performed in 15 human females and 3 human males with typical manifestation of complete stroke (12 cases) or transient ischemic attack (6 cases). The mean SP level in the serum of patients with transient ischemic attack (0.53+/-0.25 ng/ml) and of patients with complete stroke (0.31+/-0.14 ng/ml), showed significantly higher values than in controls (0.10+/-0.02 ng/ml). Moreover, in transient ischemic attack, the SP values were significantly higher than in cerebral complete stroke. But SP levels, based on the timings of classification of patients (i.e. before 12 hours: 0.34+/ 0.15 ng/ml vs. 12 to 24 hours: 0.26+/-0.11 ng/ml) with brain injury, did not show any significant difference. Both values anyway were significantly higher than in controls. Our original results demonstrate the SP increase during cerebral ischemia. Further studies are necessary to verify if SP has an effective physiopathological role in the neurological ischemic damage, or if it is only a concomitant phenomenon. Our data, if confirmed, will be particularly important, not only to improve the knowledge of cerebral ischemic injury, but also for diagnosis and therapeutic approaches. PMID- 12578733 TI - Antiphosphatidylinositol antibody in deep venous thrombosis patients. AB - Antiphospholipid antibodies are a heterogeneous group of immunoglobulins with specificity for a number of phospholipids, phospholipid-binding proteins and phospholipid-protein complexes. The association between antiphospholipid antibodies and a variety of pathologic disorders, such as arterial and venous thrombosis and recurrent pregnancy loss is recognized as Antiphospholipid Syndrome. The immunoassay currently used to detect antiphospholipid antibodies is the anticardiolipin test. Anticardiolipin antibodies are believed to be polyspecific antibodies that cross-react with all the anionic phospholipids. Therefore, testing only for anticardiolipin antibodies does not always permit detection of all antiphospholipid antibodies, specially when only IgG are evaluated. In a selected population of 74 idiopathic and secondary deep venous thrombosis patients, IgG anticardiolipin, antiphosphatidylinositol and antiphosphatidylserine antibodies were detected by solid-phase immunoassays. Our results show that by testing for each antiphospholipid family, many patients, not evidenced by the standard anticardiolipin assay, were found to be antiphospholipid-positive. The anticardiolipin positive patients have always low, moderate or high levels of antiphospholipid antibodies, suggesting that the antiphospholipid positivity is predictive of anticardiolipin positivity. It should be noted that the patients with only antiphosphatidylinositol positive antibody have a story of nervous system pathology. The meaning of these results is at present under discussion. PMID- 12578735 TI - Long-term and preventive effects of sublingual allergen-specific immunotherapy: a retrospective, multicentric study. AB - There is now an increasing body of evidence to support the practice of allergen specific sublingual-swallow immunotherapy (SLIT) in the treatment of IgE-mediated respiratory allergies. Recent studies on traditional injection therapy have pointed out that this form of treatment is not only capable to decrease actual allergic symptoms, but may also have long-term clinical and preventive effects and may influence atopy natural history. In the year 2000, our group published a retrospective, multicenter study showing the efficacy and safety of SLIT in a survey of 302 patients. We now carried out a second study on the same patients, with the aim of investigating long-term and preventive effects of SLIT. Beside the well-known safety and efficacy of this treatment (80.8% of patients reported clinical benefits), SLIT proved also to elicit long term clinical effects: over a mean follow-up of 11.6 months after the end of treatment, 80.8% of patients still maintained the previously achieved benefits. During the follow-up period, only 1% of non-asthma patients reported an onset of respiratory symptoms, and only 9.6% of patients undergoing new skin tests showed new sensitizations. All the clinical benefits were strongly linked to the length of treatment: patients with long lasting benefits were treated for a mean length of 29.1 months, while patients showing a return to pre-SLIT condition were treated for a mean 13.3 months. SLIT can obtain long-term and preventive effects so far attributed to injection immunotherapy. PMID- 12578737 TI - Skin reactions due to anti-epileptic drugs: several case-reports with long-term follow-up. AB - In this study, the clinical findings and management of allergic skin reactions induced by the most used antiepileptic drugs, Lamotrigine (LMT) and Carbamazepine (CBZ), were evaluated. Lamotrigine is an antiepileptic drug recently released in several countries; it is effective for a variety of seizure types in adults and children, both as an add-on agent and in monotherapy, and it is generally well tolerated. Clinical and epidemiologic evidence suggest serious cutaneous reactions to antiepileptic drugs are more likely to occur during the first 8 weeks and they appear to increase when drugs are administered with other anticonvulsants, such as Valproate (VPA). We selected 10 patients who presented an idiosyncratic skin rash when treated with carbamazepine (8 patients) and lamotrigine (2 patients) administered as monotherapy, and we followed up on these patients for several years. Seven reactions were mild/severe cutaneous eruptions; one Toxic Epidermal Necrolysis, a case of Stevens-Johnson and a case of Hypersensitivity Syndrome. All severe skin drug reactions were induced by Carbamazepine. In five patients the AEDs were ceased abruptly (sometimes with the administration of a different molecule), tapered in four and continued unchanged in one. We conclude that the discontinuation of the drug with substitution with another is the most effective treatment and that corticosteroids are helpful in mild cutaneous reactions, while in severe skin reactions, such as Toxic Epidermal Necrolysis, corticosteroids are only a complementary therapy since intravenous immunoglobulins are the first choice treatment. PMID- 12578736 TI - -Dual nucleoside therapy for HIV infection: analysis of results and factors influencing viral response and long term efficacy. AB - We performed a retrospective analysis of our experience with dual nucleoside regimens to look for predictors of long term benefit. The study evaluated a cohort of 68 HIV-infected patients treated at 3 Italian hospital-based facilities. The results were evaluated using univariate and multivariate statistical analysis. Fourty-three males and 25 females were treated for 22 +/- 14 months. Sixty three patients (92.6%) suffered no or low-grade side-effects. Thirty-four patients (50%) reached a viral load <400 copies/ml (undetectable). Viremia remained persistently undetectable in 9 cases (13.2%). Variable relapses of viremia were seen in 13 patients (19.1%) even though their therapys were not modified. Eight patients (11.8%) showed relapsing viremias persistently around or below 10,000 copies/ml. All patients reaching undetectable viremia but one showed increasing or stable CD4+ cell counts. Factors predicting favourable response were: pre-treatment CD4+ T-cells >150/microl, pre-treatment viremia <50,000 copies/ml, pre-treatment lymphocytes >1,500/microl, and no previous exposure to NRTI. Total lymphocyte counts and CD4+ T-cells showed a significant correlation. Dual NRTI regimens may be still considered for patients unable to tolerate HAART regimens and presenting with favourable predictors of response. PMID- 12578738 TI - The value of data. PMID- 12578739 TI - Companion animal adoption study. AB - To better understand the outcomes of companion animal adoptions, Bardsley & Neidhart Inc. conducted a series of 3 surveys over a 1-year period with dog and cat owners who had adopted their pet through either a (a) Luv-A-Pet location, (b) Adopt-a-thon, or (c) traditional shelter. This article suggests opportunities to improve owners' perceptions of their pets and the adoption process through (a) providing more information before adoption about pet health and behaviors, (b) providing counseling to potential adopters to place pets appropriately, and (c) educating adopters to promote companion animal health and retention. Results demonstrate that the pet's relationship to the family unit, such as where the pet sleeps and how much time is spent with the pet, is related to the amount of veterinary care the companion animal receives, and to long-term retention. Satisfaction and retention are attributed to the pet's personality, compatibility, and behavior, rather than demographic differences among adopters or between adoption settings. The age of the companion animal at adoption, the intended recipient, and presence of children in the home also play a role. Health problems were an issue initially for half of all adopted pets, but most were resolved within 12 months. Roughly one fourth of adopters who no longer have their companion animal said their pet died. Characteristics of pets that died support the contention that spaying and neutering profoundly affects a companion animal's life span. Although retention is similar for dogs and cats, mortality is higher among cats in the first year after adoption. PMID- 12578740 TI - Effects of lameness of dairy cows. AB - Lameness is one of the most important welfare problems in dairy cattle. Most studies on lameness have focused on wide ranging surveys to identify causal factors, but few have considered the welfare implications of this disorder. In this study, we compared the social and individual behavior of 10 lame cows and 10 nonlame cows. The 20 Holstein-Friesian cows calved in the summer and spent the autumn and winter together with another 36 nonlame cows in a Newton Rigg cubicle house building. The cubicle to cow ratio was 1:1, and wheat straw bedding was provided every day. The investigators fed the cows ad lib a silage-based diet and milked them twice a day, at which time they received adjusted amounts of concentrate. The investigators observed the 2 groups of cows a total of 32 hr to obtain information on social and individual behaviors through scan and behavior sampling. Although lame cows were less likely to start an aggressive interaction, there were no differences in times receiving aggression. No differences were found in the times licking other cows; however, the frequency of times being licked was higher in the lame cows. The lame cows spent more time lying out of the cubicles, had longer total lying times, and spent less time feeding. The behavioral differences seen show that lame cows do not cope as successfully with their environment as do nonlame cows. Also, these results provide useful information on how licking in dairy cows may play a role in alleviating discomfort of other herd members who are in pain or who are sick. PMID- 12578741 TI - Body condition of feral cats and the effect of neutering. AB - Considerable debate exists regarding the most appropriate methods for controlling feral cat populations, both from humane and logistical points of view. The physical condition of feral cats has not been reported, and it is not known if these cats benefit from neutering. This study investigates the body condition of feral cats by measuring body weight (BW), body condition score (BCS; Burkholder, 2000; Laflamme, Kealy, & Schmidt, 1994), and falciform fat pad. The study includes lateral abdominal radiographs taken at the time of neutering of 105 adult feral cats for measurement of falciform fat pad depth and area. At that time we also assessed BW and BCS. One year later we assessed the effects of neutering on body condition by evaluating a subsample of 14 cats. At the time of surgery, the cats were lean but not emaciated (BW 3.1 +/- 0.9 kg; BCS 4 +/- 1; based on a 1 to 9 scale ranging from 1 [emaciated] to 9 [grossly obese]). Falciform fat pad depth and area averaged 7.1 mm and 197.4 mm2, respectively, indicating a small amount of fat. Fourteen cats, reevaluated 1 year after neutering, increased 260% + 90% in falciform fat pad depth, 420% +/- 390% in fat pad area, 40% +/- 4% in BW, and 1 level in BCS ranking (1 to 9 scale; all differences p <.001). Similar to confined socialized cats, feral cats gained significant weight and body fat after neutering. PMID- 12578742 TI - Effects of pool size on free-choice selections by atlantic bottlenosed dolphins at one zoo facility. PMID- 12578743 TI - A project to clone companion animals. PMID- 12578744 TI - Cloning companion animals is wrong. PMID- 12578745 TI - New dog: old tricks. PMID- 12578746 TI - Hawthorne's Rebuttal. PMID- 12578747 TI - Artificial weaning of calves: benefits and costs. PMID- 12578748 TI - Comparison between color duplex ultrasonography and histology of the temporal artery in cranial arteritis (giant cell arteritis). AB - BACKGROUND: Duplex sonography of the temporal artery may be helpful in the diagnosis of cranial arteritis. PATIENTS AND METHODS: The superficial temporal arteries of 36 patients with cranial arteritis or suspected arteritis were examined using both duplex ultrasonography (US) and biopsy. The data of these patients were divided into two groups. Group A consisted of 24 patients (66.7%) with definite positive results using duplex (US) and Group B of 12 patients (33.3%) who showed a suspicious or negative ultrasonographic result. RESULT: In all patients of Group A, the histological findings corresponded with the ultrasonographic changes in the inflamed artery. - The characteristic ultrasonographic sign was a dark halo around the lumen of the temporal arteries. There was a high correlation between a bilateral halo found by US with an ocular involvement. Ten out of 14 patients with a bilateral halo (71.4%) showed a distinct involvement of the optic nerve or retina. - The characteristic histological signs were infiltration of the vessel wall by inflammatory cells, mainly lymphocytes. Group B: The biopsies of the superficial temporal arteries were positive in 8 patients (66.7 %), negative in 4 other patients (33.3%). CONCLUSION: Patients with a distinct halo, demonstrated by US, also showed corresponding pronounced inflammatory cell infiltration of the vessel wall. Patients with no ultrasonographic changes presented histological signs of initial inflammation such as isolated inflammatory cells around the vasa vasorum and/or in the adventitial layer. PMID- 12578749 TI - Women Living with HIV Does Sex and Gender Matter? A Current Literature Review. AB - BACKGROUND: As women are more susceptible to heterosexual HIV infection than men, it is predicted that the percentage of women amongst people living with HIV will continue to grow worldwide. This has not only impact on preventive and reproductive issues, but also on treatment of women with HIV. - METHODS: Abstracts from the 9 superset th Conference on Retroviruses and Opportunistic Infections in Seattle and the XIV World AIDS Conference in Barcelona in 2002 have been reviewed for this article, complemented by further internet-search. - RESULTS: Increased time was dedicated to sex and gender aspects in major conferences. Sex-mediated differences in viral replication tend to disappear within the first five to six years of HIV-infection. Providing adequate medical care, women do respond equally well or even better to antiretroviral treatment and do not progress faster than men to AIDS. Gaps in our knowledge on efficacy, side effects, and pharmacocinetics of antiretrovirals and psychosocial factors promoting adherence to treatment in women have been identified. - CONCLUSIONS: If women die faster of AIDS, it is a matter of gender, but not sex. Providing access to treatment, psychosocial and behavioural aspects of adherence are mediated as a function of gender. Missing data on side effects and pharmacokinetic profiles of various treatments in women may be explained by the lack of regulatory requirements for statistically meaningful participation of women and sex-specific analysis as licensing criteria for drug approval. Involvement of women s community advisory boards (CABs) in the design of future studies may help to gather knowledge for recommendations in the treatment of women living with HIV. PMID- 12578750 TI - Inhibition of TNF-alpha mediated cell death by HIV-1 specific protease inhibitors. AB - HIV-1 protease inhibitors have contributed significantly to the reduction in morbidity and mortality associated with HIV-1 disease. Some of their clinical benefit may be attributed to inhibition of non-viral pathogen proteases and mammalian proteases involved in apoptosis. Our objective was to investigate the effect of HIV-1 protease inhibitors on two different mechanisms of apoptosis in cells not exposed to HIV-1. Modulation of apoptosis induced in U937 or Jurkat cells by CD95 (Fas-ligand) or TNF-alpha was measured using flow cytometry using the 7-AAD and annexin/propidium iodide methods. - HIV-1 protease inhibitors reduced TNF-alpha mediated cell death in a dose-dependent manner, with a maximum inhibition ranging between 38% and 60% observed for 100 microM indinavir. Saquinavir and ritonavir, but not nelfinavir also inhibited TNF-alpha induced cell death. Nevirapine (an HIV-1 reverse transcriptase inhibitor) showed no effect. The TNF-alpha activity was also inhibited by the caspase inhibitors Z-VAD fmk at concentrations of 10 microM or less, and by DEVD-cmk. In contrast, HIV-1 protease inhibitors did not affect CD95 induced apoptosis in Jurkat cells at any of the concentrations tested. Our findings indicate that HIV-1 protease inhibitors may act on mammalian proteases involved in the regulation of apoptosis; whether this is relevant in the clinical setting remains to be established. Identification of the pathways involved may lead to a better understanding of the clinical impact of this drug class and their role in HAART associated toxicities. PMID- 12578751 TI - Effects of selected immunouppressive drugs on prostaglandin release, protein synthesis and cell proliferation in human gingival fibroblasts and on the growth of plaque bacteria. AB - BACKGROUND: Immunosuppressants play an essential role in transplantation therapy. In view of the side effects, e.g. gingival overgrowth, the present in vitro study was performed in order to investigate the effect of selected immunosuppressants on metabolic activities of gingival fibroblasts. Furthermore, the effect on the growth of six oral microorganisms was investigated. METHODS: Human gingival fibroblasts were incubated in the presence of azathioprine (Aza), cyclosporin A (CsA), tacrolimus (Tac) or mycophenolatmofetil (Myc). PGE subset 2 release was determined by means of a specific competitive enzyme immunoassay, using monoclonal antibodies specific for PGE subset 2 (clone E2R1). The protein content was measured spectrophotometrically. A redox indicator system was employed to assess the proliferation activity. In an additional trial the growth of six strains of oral bacteria (A. viscosus T14V, S. oralis H1, S. mutans 10449, C. gingivalis DR2001, A. actinomycetemcomitans Y4, and M. micros 33270) in the presence of the immunosuppressants was measured. RESULTS: In comparison with the controls, the PGE subset 2 release was increased by 39.3% following incubation with Aza, and by 77.0% with CsA. The protein concentrations (1 g immunosuppressant / ml medium) were reduced by 26.0% for Aza and 17.0% for Myc. Furthermore, a drug-dependent inhibition in the cell proliferation rate was noted after an incubation period of 6 hours (Aza 70.7%, CsA 78.2%, Myc 69.8%, Tac 64.0%). The most pronounced growth-inhibiting effects were observed for CsA at values ranging from 21.0% (S. mutans 10449) to 48.6% (A. viscosus T14V) growth inhibition. CONCLUSIONS: The present study with common immunsuppresants demonstrated both a medication- and dose-dependent alteration in the metabolic activity of gingival fibroblasts. Furthermore, growth-inhibitory effects on the selected bacterial strains could be observed. PMID- 12578752 TI - Effect of periodontal therapy on sulcular sulphide level a longitudinal study. AB - PURPOSE: The identification of active disease sites is a leading goal in basic periodontal research. Of toxic bacterial metabolites detectable in gingival crevicular fluid, volatile sulphur compounds (VSC) have been implicated in periodontal tissue destruction. Several bacteria associated with active destructive disease are capable of producing VSC, this fact supporting the idea of sulcular VSC being a possible marker of disease activity. A new portable sulphide monitor providing chairside information on sulcular sulphide level (SU) has been developed. The aim of this study was 1) to monitor the effect of mechanical therapy on SU and clinical parameters and 2) to clarify whether SU measurements might have the potential to detect disease activity. METHODS: 34 patients (22 M, 12 F) with generalized or localized chronic periodontitis received periodontal treatment in a private practice consisting of an oral hygiene phase (HP) lasting several weeks, scaling and root planing (SRP), and flap surgery at sites >5 mm or with furcation involvement. Subjects were examined three times (1 week after the diagnosis was made, at the end of HP and at the 1st maintenance session 3 months after SRP) recording clinical parameters (clinical attachment loss CAL, probing depth PD, bleeding on probing intensity BI, plaque index PI) and sulcular sulphide level (SU) measured by the portable monitor as as digital score ranging from 0.0 (<10 superset 6 M of S superset 2-) to 5.0 (10 superset 2 M of S superset 2-) in increments of 0.5. RESULTS: 23 patients (15 M, 8 F, 47.5 +/- 9 years) were included in the data analysis. Periodontal therapy resulted in a significant reduction of mean BI by 0.69 +/- 0.45, of mean PD by 1.39 +/- 0.33 mm and in a mean gain of attachment of 1.07 +/- 0.38 mm (p = 0.0001). The clinical improvement was accompanied by a reduction of mean SU by 0.20 +/- 0.13 and of the mean percentage of SU-positive sites per patient (SUp) by 20.09 +/- 13. SU-positive sites were located at all types of teeth. 67.9 % of SU-positive sites and 83.8 % of sites with a SU > 1 were found at the molars. 16.1 % of initially 579 SU-positive sites remained SU-positive. For these sites, BI reduction by treatment was significantly lower compared with SU-negative sites (p < 0.01). SU was significantly correlated with PD, CAL, BI and PI (p = 0.0001). Sensitivity and specificity of SU related to BOP (BI > 0) was 29.3 % and 96.8 %, respectively. CONCLUSIONS: Sulcular VSC as measured by the portable sulphide monitor may be an indicator of therapeutical success and periodontal stability. The sensor system offers chairside information on sulcular sulphide production at specific sites and is a valuable supplementation of traditional clinical methods in periodontal examination. More controlled longitudinal studies are needed to clarify the role of sulcular VSC at sites undergoing periodontal destruction. PMID- 12578754 TI - Pupillary escape. PMID- 12578756 TI - Adverse events in cataract surgery. PMID- 12578758 TI - Medical vs surgical therapy in preventing visual field loss. PMID- 12578760 TI - Medical vs surgical therapy in preventing visual field loss. PMID- 12578761 TI - Medical vs surgical therapy in preventing visual field loss. PMID- 12578762 TI - No colobomas in "renal coloboma" syndrome. PMID- 12578764 TI - Vision loss: a public health problem? PMID- 12578765 TI - U.S. Food and Drug Administration clinical trial of the Implantable Contact Lens for moderate to high myopia. AB - PURPOSE: To assess the safety and efficacy of the Implantable Contact Lens (ICL) to treat moderate to high myopia. DESIGN: Prospective nonrandomized clinical trial. PARTICIPANTS: Five hundred twenty-three eyes of 291 patients with between 3 and 20.0 diopters (D) of myopia participating in the U. S. Food and Drug Administration clinical trial of the ICL for myopia. INTERVENTION: Implantation of the ICL. MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA), refraction, best spectacle-corrected visual acuity (BSCVA), adverse events, operative and postoperative complications, lens opacity analysis (Lens Opacity Classification System III), subjective satisfaction, and symptoms. RESULTS: Twelve months postoperatively, 60.1% of patients had a visual acuity of 20/20 or better, and 92.5% had an uncorrected visual acuity of 20/40 or better. Patients averaged a 10.31-line improvement in UCVA, 61.6% of patients were within 0.5 D, and 84.7% were within 1.0 D of predicted refraction. Only one case (0.2%) lost > 2 lines of BSCVA. Gains of 2 or more lines of BSCVA occurred in 55 cases (11.8%) at 6 months and 41 cases (9.6%) at 1 year after ICL surgery. Early and largely asymptomatic, presumably surgically induced anterior subcapsular (AS) opacities were seen in 11 cases (2.1%); an additional early AS opacity (0.2%) was seen because of inadvertent anterior chamber irrigation of preservative-containing solution at surgery. Two (0.4%) late (> or = 1 year postoperatively) AS opacities were observed. Two (0.4%) ICL removals with cataract extraction and intraocular lens implantation have been performed. Patient satisfaction (very/extremely satisfied) was reported by 92.4% of subjects on the subjective questionnaire; only four patients (1.0%) reported dissatisfaction. Slightly more patients reported an improvement at 1 year over baseline values for the following subjective symptoms: quality of vision, glare, double vision, and night driving difficulties. Only a 3% difference between pre-ICL and post-ICL surgery was reported for haloes. CONCLUSIONS: The results support the safety, efficacy, and predictability of ICL implantation to treat moderate to high myopia. PMID- 12578766 TI - Risk factors and prognosis for corneal ectasia after LASIK. AB - PURPOSE: To review cases of corneal ectasia after laser in situ keratomileusis (LASIK), identify preoperative risk factors, and evaluate methods and success rates of visual rehabilitation for these cases. DESIGN: Retrospective nonrandomized comparative trial. PARTICIPANTS: Ten eyes from seven patients identified as developing corneal ectasia after LASIK, 33 previously reported ectasia cases, and two control groups with uneventful LASIK and normal postoperative courses: 100 consecutive cases (first control group), and 100 consecutive cases with high myopia (> 8 diopters [D]) preoperatively (second control group). METHODS: Retrospective review of preoperative and postoperative data for each case compared with that of previously reported cases and cases with uneventful postoperative courses. MAIN OUTCOME MEASURES: Preoperative refraction, topographic features, residual stromal bed thickness (RSB), time to the development of ectasia, number of enhancements, final best-corrected visual acuity (BCVA), and method of final correction. RESULTS: Length of follow-up averaged 23.4 months (range, 6-48 months) after LASIK. Mean time to the development of ectasia averaged 16.3 months (range, 1-45 months). Preoperative refraction averaged -8.69 D compared with -5.37 D for the first control group (P = 0.005). Preoperatively, 88% of ectasia cases met criteria for forme fruste keratoconus, compared with 2% of the first control group (P < 0.0000001) and 4% of the second control group (P = 0.0000001). Seven eyes (70%) had RSB <250 microm, as did 16% of eyes in the first control group and 46% of the second control group. The mean RSB for ectasia cases (222.8 microm) was significantly less than that for the first control group (293.6 micro m, P = 0.0004) and the second control group (256.5 microm; P = 0.04). Seven eyes (70%) had enhancements. Only 10% of eyes lost more than one line of BCVA, and all patients eventually achieved corrected vision of 20/30 or better. One case required penetrating keratoplasty (10%), while all others required rigid gas-permeable contact lenses for correction. CONCLUSIONS: Significant risk factors for the development of ectasia after LASIK include high myopia, forme fruste keratoconus, and low RSB. All patients had at least one risk factor other than high myopia, and significant differences remained even when controlling for myopia. Multiple enhancements were common among affected cases, but their causative role remains unknown. We did not identify any patients who developed ectasia without recognizable preoperative risk factors. PMID- 12578767 TI - An outbreak of Mycobacterium chelonae infection after LASIK. AB - OBJECTIVE: To describe an outbreak of mycobacterial keratitis after laser in situ keratomileusis (LASIK), including the microbiologic investigation, clinical findings, treatment response, and outcome. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Patients (n = 10) who underwent LASIK surgery between August 22 and September 4, 2000, and developed mycobacterial infection. METHODS: Patients were prospectively followed in relation to microbiologic investigation, clinical findings, treatment response, and outcome. MAIN OUTCOME MEASURES: Most patients underwent bilateral simultaneous LASIK. Postoperative infection was signaled by the appearance of corneal infiltrates in the third postoperative week. The microbiologic workup was performed on cultures obtained either by direct scraping of the cornea or by lifting the flap. Medical therapy was instituted based on drug susceptibility testing. Surgical interventions such as corneal debridement and flap removal were performed during recurrences or when there was no satisfactory clinical response. RESULTS: Cultures revealed Mycobacterium subspecies chelonae. Patients were treated with topical clarithromycin (1%), tobramycin (1.4%), and ofloxacin (0.3%). Oral clarithromycin (500 mg twice a day) was prescribed for those patients who did not respond clinically to topical treatment. Four eyes healed on this regimen. Flap removal was necessary in seven eyes. CONCLUSIONS: This report highlights mycobacteria as an etiologic infectious agent after LASIK. Diagnosis can be difficult and is often delayed. The treatment mainstay is prolonged antibiotic therapy. Surgical debridement and flap removal may shorten the disease course. PMID- 12578768 TI - Prediction of visual outcome after penetrating keratoplasty for pseudophakic corneal edema. AB - OBJECTIVE: To evaluate the ability to predict visual outcome after penetrating keratoplasty (PKP) in patients with pseudophakic corneal edema (PCE) or aphakic corneal edema (ACE) based on preoperative parameters available from the patient history and ocular examination. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Forty-eight patients who underwent 59 PKP procedures for PCE between 1997 and 2000 by two cornea specialists. METHODS: Medical records were retrospectively analyzed for variables in the history and ocular examination before PKP and visual outcome after PKP. Variables included age, gender, presence of diabetes or cardiovascular disease, method of intraocular lens (IOL) implantation during cataract surgery, vitreous loss during cataract surgery, time between cataract and PKP surgery, and maximal visual acuity reached after cataract surgery. The predictive value of each preoperative variable on post-PKP visual outcome was assessed using both univariate and multiple regression analyses. MAIN OUTCOME MEASURES: Statistical significance for the predictive value of each preoperative variable on post-PKP visual outcome. RESULTS: Best corrected visual acuity (BCVA) of 20/40 or better was achieved in 13 patients (27%). The strongest predictor of this outcome was implantation of a bag-fixated or sulcus-fixated IOL at the time of cataract surgery (P = 0.007; odds ratio, 15.8; 95% confidence interval, 1.2-208). Less significant variables included BCVA after cataract surgery, time between cataract surgery and PKP, and gender. CONCLUSIONS: In planning and advising patients with pseudophakic or aphakic corneal edema who are candidates for PKP, the method of IOL implantation during the cataract surgery is the single most significant predictor of visual acuity after corneal transplantation. Bag-fixated or sulcus-fixated posterior chamber IOL was associated with a better visual outcome than anterior chamber IOL, scleral-fixated posterior chamber IOL, or aphakia. PMID- 12578769 TI - Fibrin glue versus N-butyl-2-cyanoacrylate in corneal perforations. AB - OBJECTIVE: To compare the efficacy of fibrin glue and N-butyl-2-cyanoacrylate in corneal perforations. DESIGN: Randomized, controlled clinical trial. PARTICIPANTS: Forty-one patients (41 eyes) with corneal perforations up to 3 mm in diameter with a positive Seidel's test were randomly assigned to two groups (1 and 2). INTERVENTION: Group 1 comprised 19 eyes treated with fibrin glue, and group 2 comprised 22 eyes treated with N-butyl-2-cyanoacrylate. MAIN OUTCOME MEASURES: Number of eyes with successful healing, time required for healing, status of corneal vascularization, and complications were compared in the two groups. Power calculation was performed at alpha = 0.05. RESULTS: Fifteen (79%) eyes had successful healing of corneal perforation in group 1, compared with 19 (86%) eyes in group 2 (P > 0.05) at 3 months' follow-up. The power to detect a difference between the two groups was 10%. Corneal perforation healed within 6 weeks in 12 (63%) eyes in group 1 and 7 (31.8%) eyes in group 2 (P < 0.05). Reapplication of glue was required in six (31.5%) eyes in group 1 and seven (31.4%) eyes in group 2 during the first 3 months of follow-up. The mean number of applications per eye was 1.37 in group 1 and 1.36 in group 2. An increase in deep corneal vascularization was observed in 2 (10.5%) eyes in group 1 and 10 (45.5%) eyes in group 2 (P < 0.05). Giant papillary conjunctivitis occurred in one (5%) eye in group 1 and eight (36.4%) eyes in group 2 (P < 0.05). CONCLUSIONS: Fibrin glue and cyanoacrylate tissue adhesive are both effective in the closure of corneal perforations up to 3 mm in diameter. Fibrin glue provides faster healing and induces significantly less corneal vascularization, but it requires a significantly longer time for adhesive plug formation. PMID- 12578770 TI - Contact lenses for the treatment of pediatric cataracts. AB - OBJECTIVE: To evaluate the experiences, attitudes, and perceptions of the caregivers of children with cataracts who were visually rehabilitated with contact lenses. PARTICIPANTS: One hundred twenty-three caregivers of children <8.1 years old treated for unilateral and bilateral cataracts at one pediatric hospital. DESIGN: Survey by questionnaire. INTERVENTION: Primary caregivers were asked to complete an anonymous questionnaire. MAIN OUTCOME MEASURES: Caregiver responses to questions assessing background and demographic and clinical information, as well as perceptions, attitudes, levels of compliance, and anxiety with respect to treatment, were reviewed. Caregivers were also asked to choose between aphakic rehabilitation with contact lenses, aphakic glasses, or intraocular lenses, given various hypothetical scenarios differing in regard to their final visual prognosis, risks of treatment complications, and cost. RESULTS: The response rate was 82.9%. Absolute average stress levels for contact lens use were 1.36 +/- 1.79 and 0.79 +/- 1.48 (scale, 0-5) for insertion and removal, respectively, compared with 4.03 +/- 1.64 and 2.40 +/- 1.92 for cataract surgery and patching therapy, respectively. Although average paired initial resistance to treatment (RT) levels for contact lens insertion and removal on a scale of 0 to 3 were high (2.09 +/- 1.15) and moderate (1.63 +/- 1.20), respectively, final RT levels were significantly lower (1.09 +/- 1.14 and 0.66 +/ 1.07, respectively; P < 0.0001). The vast majority of caregivers chose contact lens use in hypothetical scenarios that depicted realistic expectations for other forms of aphakic rehabilitation. CONCLUSIONS: In our study, contact lenses seemed to be well tolerated by most patients, as assessed by caregivers. Although initial resistance to contact lens use is high, this decreases with time. Relative to other events in the treatment of pediatric cataracts, contact lens use is not a major stressor for most caregivers and patients. This study supports the notion that contact lenses should continue to receive serious consideration as a treatment option for pediatric cataracts. PMID- 12578771 TI - Silicone nasolacrimal intubation with mitomycin-C: a prospective, randomized, double-masked study. AB - OBJECTIVE: To determine if mitomycin-C (MMC) application during silicone intubation (SI) can effectively substitute for dacryocystorhinostomy (DCR) or Jones tube insertion. DESIGN: A prospective, randomized, double-masked study. PARTICIPANTS: Twenty-four patients (7 males, 17 females; aged 2-69 years) with 27 eyes (three bilateral cases) with an obstructed lacrimal system who were candidates for DCR or Jones tube. METHODS: All study patients underwent SI with application in a randomized, double-masked fashion of MMC or placebo, with the former receiving 0.2 mg/ml MMC for 2 minutes before SI. MAIN OUTCOME MEASURES: Clinical assessment of tearing and discharge; Schirmer I and II tests at 1, 3, 6, and 12 months; and the need for eventual DCR or Jones tube insertion. RESULTS: Follow-up ranged from 13 to 23 months (mean, 18 months; median, 17 months). Seven of the 12 eyes in the MMC group and 8 of the 15 eyes in the placebo group had a successful outcome and remained symptom free. The difference was not statistically significant (P = 0.79). Dacryocystorhinostomy or Jones tube was performed in five eyes in the MMC group and in seven eyes in the placebo group 3 to 18 months later; all had successful outcome and no complications. Untoward effects of MMC application included a slit canaliculus and a transient conjunctival injection. A successful outcome using this combined technique does not appear to correlate with age, gender, laterality, and intraoperative bleeding observed at the punctum or for the duration of symptoms. CONCLUSIONS: Mitomycin-C application during SI did not benefit outcome. Complications from such application were mild and infrequent. PMID- 12578772 TI - Monitored anesthesia care for enucleations and eviscerations. AB - OBJECTIVE: To report the technique and success of using monitored anesthesia care instead of general anesthesia for ocular enucleation and evisceration surgeries. DESIGN: Retrospective, noncomparative interventional case series. PARTICIPANTS: Twelve enucleated patients (Soparkar and Patrinely) and 146 eviscerated patients (Kulwin and Kersten). METHODS: Surgical logs of two oculoplastic practices were reviewed searching for cases of ocular enucleations and eviscerations performed under monitored anesthesia care between 1990 and 2001. Identified hospital and clinic charts were then reviewed. MAIN OUTCOME MEASURES: Monitored anesthesia care was deemed successful if (1) there were hemodynamic stability and complete analgesia intraoperatively; (2) there was absence of any chart documentation regarding patient or family psychological distress over the anesthesia method used; and (3) patients were discharged from the hospital without the need for observation or treatment > or = 23 hours. RESULTS: Between 1990 and 2001, 146 eviscerations were performed under local anesthesia with monitored anesthesia care by two surgeons (RCK, DRK) as their routine practice pattern. In 1996, Drs. Soparkar and Patrinely began performing enucleations under monitored anesthesia care in selected cases, and from 1996 to 2001, these surgeons enucleated 12 patients under monitored anesthesia care. Four of the enucleated patients requested surgery without general anesthesia. The remaining eight patients had been refused surgery by at least one other specialist because of the patient's perceived high medical risk for complications under general anesthesia. In all 158 patients, the procedures were deemed successful by the preceding criteria. CONCLUSIONS: This four-surgeon case series reports the successful use of local anesthesia with monitored care for ocular enucleation and evisceration procedures, offering several potential advantages over the traditional use of general anesthesia. PMID- 12578773 TI - Malignant mesenchymoma of the orbit: case report and review of the literature. AB - OBJECTIVE: Malignant mesenchymoma are rare tumors of the orbit. From 1961 using English-language literature, we present the sixth such case and the first case with three malignant components. DESIGN: Interventional case report. METHODS: The clinical presentation, workup, surgical treatment, and pathology of a case of malignant mesenchymoma of the orbit are presented. RESULTS: Although very rare, these tumors should be in the differential diagnosis of any tumor of the orbit. The prognosis is unknown because of the lack of follow-up of reported patients. CONCLUSIONS: This malignant orbital mesenchymoma, an entity not accepted by all pathologists, was more complex than cases previously reported in the English literature in demonstrating rhabdomyosarcomatous, chondrosarcomatous, and osteogenic differentiation. PMID- 12578774 TI - Metastatic gastroesophageal junction adenocarcinoma to the extraocular muscles. AB - OBJECTIVE: To present one definitive and one probable case of gastroesophageal adenocarcinoma metastases to the extraocular muscles. DESIGN: Two observational case reports. METHODS: Two interventional case reports with clinical pathologic correlation and review of the literature. Vanderbilt University Institutional Review Board approval was obtained for this report. MAIN OUTCOME MEASURES: We analyzed the presenting symptoms, visual acuity, motility, and external examination. RESULTS: Computed tomography showed nodular enlargement of an isolated rectus muscle. Histopathologic examination confirmed the diagnosis of metastatic gastrointestinal carcinoma in one case. The orbital neoplasm contained signet ring cells, classically seen in gastrointestinal malignancy. CONCLUSIONS: Metastasis from gastrointestinal carcinoma should be considered in the differential diagnosis of a patient presenting with diplopia or visual loss, particularly with a recent history of gastrointestinal malignancy. PMID- 12578775 TI - Gaze-evoked amaurosis: a report of five cases. AB - OBJECTIVE: To highlight the various causes of gaze-evoked amaurosis. DESIGN: Retrospective noncomparative interventional case series. PARTICIPANTS: Five patients treated at our facility over the past 6 years. METHODS: Clinical presentation, radiologic studies, surgical management, and postsurgical results are presented. MAIN OUTCOME MEASURES: Visual acuity, clinical findings of gaze evoked amaurosis. RESULTS: Only two patients had classic intraorbital etiologies, one with an intraconal cavernous hemangioma and one with an intraconal foreign body. Three patients had extraorbital processes, two with orbital fractures and one with a sinus tumor. Only two of our patients initially were aware of the gaze evoked amaurosis at presentation. Appropriate surgery was curative in all cases. CONCLUSIONS: Gaze-evoked amaurosis is a rare condition, classically implicating intraconal orbital pathology. In one of the largest case series published to date, we found extraorbital etiologies are also capable of producing gaze-evoked vision loss. Gaze-evoked amaurosis should be suspected and tested for in any orbital condition. PMID- 12578776 TI - Optic disc structure and shock-induced anterior ischemic optic neuropathy. AB - PURPOSE: To describe a patient who developed unilateral shock-induced anterior ischemic optic neuropathy (SIAION) after gastrointestinal hemorrhage followed by presumed idiopathic nonarteritic anterior ischemic optic neuropathy (NAION) in the fellow eye. DESIGN: Retrospective, observational case report and literature review. METHODS: The case history of an 80-year-old man who developed SIAION, followed by NAION in the fellow eye, was reviewed. All previously reported cases of SIAION were reviewed. MAIN OUTCOME MEASURES: Neuro-ophthalmic examination, including visual acuity, funduscopy, and automated perimetry. RESULTS: An 80-year old man, with a history of gastrointestinal bleeding from a duodenal ulcer, was hospitalized and received four units of packed red blood cells after he was found to be severely anemic (hemoglobin 6.7 g/dl). Three days later he complained of loss of vision of the right eye. Neuro-ophthalmic examination 2 weeks later disclosed a visual acuity of counting fingers at 6 inches in the right eye and 20/40 in the left eye, with a right relative afferent pupillary defect and a superior altitudinal visual field defect. Funduscopy revealed optic disc edema with a temporal parapapillary hemorrhage in the right eye and a small optic disc, with no cup, in the left eye. A diagnosis of SIAION secondary to anemia was made. Six weeks later he developed a new inferior altitudinal visual field defect in the left eye and diffuse optic disc swelling. He had no signs or symptoms of giant cell arteritis or polymyalgia rheumatica, his hemoglobin at this time was 11.9 g/dl, and the Westergren erythrocyte sedimentation rate was 6 mm/hour. CONCLUSIONS: Our patient developed optic disc swelling of the right eye after an episode of gastrointestinal hemorrhage (SIAION). The disc swelling in the left eye occurred 8 weeks later, when his hemoglobin had increased to 11.9 g/dl. The timing of the ischemic optic neuropathies suggests that the acute anemia led to involvement of the first but not the second eye. The configuration of the optic disc may have predisposed not only to the second event (NAION) but also to the first episode (SIAION). PMID- 12578777 TI - The relationship between visual disability and visual field in patients with glaucoma. AB - PURPOSE: To evaluate the relationship between visual disability and the remaining visual field in glaucoma patients. DESIGN: Prospective noncomparative survey. PARTICIPANTS: One hundred forty-seven Japanese patients with glaucoma were examined. METHODS: Using a previously developed questionnaire, we assessed the relationship between visual disability indices and both the visual field examined using program 30-2 of the Humphrey Field Analyzer and visual acuity. MAIN OUTCOME MEASURES: Pearson's correlation coefficients between visual disability indices and the 22 visual field and 2 visual acuity measurements were calculated. Multiple regression analysis with stepwise variable selection identified factors significantly contributing to visual disability. RESULTS: We identified a significant correlation between visual disability and both the visual field and visual acuity measurements collected. Of the factors examined, retinal sensitivity in the lower hemifield within 5 degrees of the fixation and visual acuity in the better eye significantly contributed to all the item and total disability indices (DIs), measurements of the degree of visual disability as evaluated by the questionnaire; visual acuity in the worse eye contributed to two of the seven item DIs, as well as the total DI. The R value for total DI (R = 0.87, P < 0.0001) indicated that approximately 76% of the visual disability could be explained by these factors. CONCLUSIONS: The Japanese glaucoma patients' visual disability is explained primarily by retinal sensitivity in the lower hemifield within 5 degrees of the fixation and visual acuity in the better eye and secondarily by visual acuity in the worse eye. PMID- 12578778 TI - Reproducibility of evaluation of optic disc change for glaucoma with stereo optic disc photographs. AB - PURPOSE: To determine the reproducibility of the assessment for glaucomatous change in serial optic disc stereo-slides. DESIGN: Masked interobserver variability study. PARTICIPANTS: Serial optic disc stereo-slides from 40 patients. METHODS: Three independent ophthalmologists evaluated for change a set of two serial 20 degrees optic disc color stereo-slides of 40 patients. This test set was not from European Glaucoma Prevention Study (EGPS) patients. Each observer performed two evaluations at least 30 days apart and was masked from the temporal sequence of the slides and his or her previous evaluation. Each patient was graded as changed or stable by two-out-of-three agreement. A kappa statistic was used to calculate the intra- and interobserver reproducibility as well as the assignment reproducibility (first consensus versus second consensus). The same procedure was followed to test the reproducibility when another experienced ophthalmologist was added to one of the three reading centers. MAIN OUTCOME MEASURES: Reproducibility in evaluating glaucomatous optic disc change. RESULTS: The intraobserver reproducibility (95% confidence interval [CI]) in the evaluation of change ranged between 0.79 (0.45-1.14) and 1.00 (0.69-1.31). The interobserver reproducibility (95% CI) in the evaluation of change ranged between 0.45 (0.15-0.75) and 0.75 (0.44-1.06). The assignment reproducibility (first consensus versus second consensus in the evaluation of change) between the senior EGPS readers was 0.94 (0.63-1.25). The assignment reproducibility when another experienced ophthalmologist replaced one of the readers was 0.94 (0.63-1.25). CONCLUSIONS: The assignment reproducibility of three expert readers looking for glaucomatous change in serial optic disc stereo-slides was excellent. It remained so when one of the three experts was replaced by another experienced reader. PMID- 12578779 TI - Clinical characteristics and leakage of functioning blebs after trabeculectomy with mitomycin-C in primary glaucoma patients. AB - PURPOSES: To describe the clinical characteristics of functioning blebs in Japanese glaucoma patients after primary trabeculectomy with adjunctive mitomycin C (MMC) and to correlate their associations with postoperative bleb leakage. DESIGN: A prospective, observational case series. PARTICIPANTS: One hundred sixty two glaucoma patients who had undergone primary trabeculectomy with MMC at the University of Tokyo Hospital at least 3 months before were examined between December 1997 and February 1998. METHODS: A predesigned data form was completed at each visit. Ophthalmologic examinations included Goldmann applanation tonometry, slit-lamp examination, and Seidel tests with and without digital ocular pressure (DOP). MAIN OUTCOME MEASURES: Properties of the functioning bleb, including bleb appearance, thickness of bleb wall, dimensions of bleb and avascular area, and leakage status with and without DOP. RESULTS: Of 162 Japanese patients, 162 eyes with functioning blebs were included. There were no differences in bleb characteristics among the different types of primary glaucoma. With a long postoperative duration, blebs tended to be thinner (P = 0.024). With DOP, the leaking rate increased from 3.1% to 5.6%, and the oozing rate increased from 11.1% to 35.8% (P < 0.001). Logistic regression analysis indicated that the larger the avascular area, the more likely the bleb leaked without DOP (P = 0.016). When DOP was applied, leakage was more likely to occur in the blebs with a long postoperative duration (P = 0.002) or with a large avascular area (P < 0.001). CONCLUSIONS: The clinical characteristics of filtering blebs were similar in Japanese patients with different types of primary glaucoma. Blebs with a large avascular area were associated with a higher risk of bleb leakage. Attention to the increased chance of leakage induced by DOP is important, especially for blebs with a long postoperative duration. Ophthalmologists should be aware of late bleb-related complications in addition to bleb function. PMID- 12578780 TI - Intraocular pressure control and complications with two-stage insertion of the Baerveldt implant. AB - PURPOSE: To evaluate intraocular pressure (IOP) control and the prevalence of complications associated with the two-stage insertion of the Baerveldt implant. DESIGN: Retrospective, noncomparative, interventional case series. METHODS: We reviewed the medical records of all patients who underwent the two-stage Baerveldt implantation for advanced glaucoma between January 1995 and November 1998 at the Doheny Eye Institute, Los Angeles, California. All eyes with a minimum of 6 months of follow-up were included in the study. Eyes that failed before 6 months after stage II were also included in the study. MAIN OUTCOME MEASURES: The criterion for surgical success was postoperative IOP of 6 mmHg or more and 21 mmHg or less on two or more consecutive measurements with or without the use of glaucoma medications and without loss of light perception or further surgical intervention to lower IOP. RESULTS: Nineteen eyes from 19 patients were included. Median postoperative follow-up was 17 months (range, 3-56 months). The cumulative probability of success was 89% at both 12 and 24 months. The median preoperative IOP of 21 mmHg (range, 12-33 mmHg) was reduced to a median postoperative IOP of 12 mmHg (range, 1-17 mmHg; P < 0.0001). The median number of glaucoma medications before surgery was three and after surgery was one (P < 0.0001). Visual acuity remained the same or improved in 13 eyes (68%). One eye experienced hypotony (5%), and in two eyes (11%) suprachoroidal hemorrhages developed. CONCLUSIONS: The two-stage insertion of the Baerveldt implant effectively reduced IOP, produced few complications, and preserved visual acuity in more than two thirds of eyes with advanced glaucomatous damage. PMID- 12578781 TI - Effect of Ginkgo biloba extract on preexisting visual field damage in normal tension glaucoma. AB - OBJECTIVE: To evaluate the effect of Ginkgo biloba extract (GBE) on preexisting visual field damage in patients with normal tension glaucoma (NTG). DESIGN: Prospective, randomized, placebo-controlled, double-masked cross-over trial. PARTICIPANTS: Twenty-seven patients with bilateral visual field damage resulting from NTG. INTERVENTION: Patients received 40 mg GBE, administered orally, three times daily for 4 weeks, followed by a wash-out period of 8 weeks, then 4 weeks of placebo treatment (identical capsules filled with 40 mg fructose). Other patients underwent the same regimen, but took the placebo first and the GBE last. Visual field tests, performed at baseline and at the end of each phase of the study, were evaluated for changes. MAIN OUTCOME MEASURES: Change in visual field and any ocular or systemic complications. RESULTS: After GBE treatment, a significant improvement in visual fields indices was recorded: mean deviation (MD) at baseline versus MD after GBE treatment, 11.40 +/- 3.27 dB versus 8.78 +/- 2.56 dB (t = 8.86, P = 0.0001, chi-square test); corrected pattern standard deviation (CPSD) at baseline versus CPSD after GBE treatment, 10.93 +/- 2.12 dB versus 8.13 +/- 2.12 dB (t = 9.89, P = 0.0001, chi-square test). No significant changes were found in intraocular pressure, blood pressure, or heart rate after placebo or GBE treatment. Any ocular and systemic side effects were recorded for the duration of the trial. CONCLUSIONS: Ginkgo biloba extract administration appears to improve preexisting visual field damage in some patients with NTG. PMID- 12578784 TI - Temba glaucoma study: a population-based cross-sectional survey in urban South Africa. AB - OBJECTIVE: To determine the prevalence and features of glaucoma in an urban South African black population. DESIGN: Random sampling cross-sectional population survey. PARTICIPANTS: Black residents of Temba, North West Province, South Africa, age > or =40 years. MAIN OUTCOME MEASURES: Automated visual field testing and detailed, standardized slit-lamp examination were attempted on all subjects. Glaucoma was diagnosed by use of the scheme proposed by the Working Group for Defining Glaucoma of the International Society of Geographical and Epidemiologic Ophthalmology on the basis of evidence of end-organ damage. RESULTS: Of 1120 subjects, 839 (74.9%) were examined. The age- and gender-adjusted prevalence of glaucoma of all types was 5.3% (95% confidence interval [CI], 3.9%-7.1%). Primary open-angle glaucoma (POAG) was the most common glaucoma diagnosis, with an adjusted prevalence of 2.9% (95% CI, 1.9%-4.3%). Secondary glaucoma occurred with an adjusted prevalence of 2.0% (95% CI, 1.2%-3.3%). Exfoliative glaucoma was responsible for 16% of all glaucoma cases. The prevalence of primary angle closure glaucoma was 0.5% (95% CI, 0.13%-1.2%). Of all subjects with glaucoma, 58% were blind in at least one eye. The prevalence of all types of glaucoma increased with age. Of subjects with POAG, 87% had not been previously diagnosed. CONCLUSIONS: The prevalence of glaucoma in this South African population was higher than that found in white populations, and most cases were undiagnosed and untreated. Glaucoma is a major cause of blindness in this population. PMID- 12578783 TI - Age, gender, biometry, refractive error, and the anterior chamber angle among Alaskan Eskimos. AB - BACKGROUND: The prevalence of angle-closure glaucoma (ACG) is greater for Eskimos/Inuit than it is for any other ethnic group in the world. Although it has been suggested that this prevalence may be due to a population tendency toward shallower anterior chamber angles, available evidence for other populations such as Chinese with high rates of ACG has not consistently demonstrated such a tendency. METHODS: A reticule, slit-lamp, and standard Goldmann one-mirror goniolens were used to make measurements in the anterior chamber (AC) angle according to a previously reported protocol for biometric gonioscopy (BG) (Ophthalmology 1999;106:2161-7). Measurements were made in all four quadrants of one eye among 133 phakic Alaskan Eskimos aged 40 years and older. Automatic refraction, dilated examination of the anterior segment and optic nerve, and A scan measurements of AC depth, lens thickness, and axial length were also carried out for all subjects. RESULTS: Both central and peripheral AC measurements for the Eskimo subjects were significantly lower than those previously reported by us for Chinese, blacks, and whites under the identical protocol. Eskimos also seemed to have somewhat more hyperopia. There were no differences in biometric measurements between men and women in this Eskimo population. Angle measurements by BG seemed to decline more rapidly over life among Eskimos and Chinese than blacks or whites. Although there was a significant apparent decrease in AC depth, increase in lens thickness, and increase in hyperopia with age among Eskimos, all of these trends seemed to reverse in the seventh decade and beyond. CONCLUSIONS: Eskimos do seem to have shallower ACs than do other racial groups. Measurements of the AC angle seem to decline more rapidly over life among Eskimos than among blacks or whites, a phenomenon also observed by us among Chinese, another group with high ACG prevalence. This apparent more rapid decline may be due to a cohort effect with higher prevalence of myopia and resulting wider angles among younger Eskimos and Chinese. PMID- 12578785 TI - Photoreceptor transplantation in retinitis pigmentosa: short-term follow-up. AB - PURPOSE: To explore the use of adult human photoreceptor transplantation as a treatment for advanced retinitis pigmentosa (RP). DESIGN: Prospective noncomparative case series. PARTICIPANTS: Eight patients with advanced RP. INTERVENTION: Transplantation of adult human cadaver photoreceptor sheets harvested with the excimer laser. No immunosuppression was used postoperatively. Patients were followed for 12 months postoperatively. MAIN OUTCOME MEASURE: Visual acuity and retinal function measured by psychophysical, electrophysiologic, and clinical testing. RESULTS: Best-corrected visual acuity (Bailey-Lovie chart), median reading speed, contrast sensitivity, and visual fields for the operated eye were not statistically significantly improved postoperatively. The amplitude and latency of the maculoscope electroretinogram, as well as the log threshold for dark adaptation, did not change between the operated and control (unoperated) eye. There was no detectable homograft reaction on slit-lamp biomicroscopy or fluorescein angiography. The only adverse effect observed was one patient who complained of monocular diplopia after retinal transplantation and subsequent cataract surgery. CONCLUSIONS: Allogeneic adult human photoreceptor transplantation is feasible in RP but was not associated with rescue of central vision or a delay in visual loss. However, any possible slowing in the rate of retinal degeneration will take many years to determine. PMID- 12578786 TI - Fundus autofluorescence and age-related macular degeneration. AB - PURPOSE: To evaluate the autofluorescence images of patients with nonexudative and the fellow eyes with exudative age-related macular degeneration (AMD). DESIGN: Observational case series. PARTICIPANTS: Fifty-four patients seen in the author's practice. METHODS: A fundus camera-based system for autofluorescence photographs was used, and the wavelengths for the excitation (580 nm) and barrier (695 nm) filters were based on known transmission and autofluorescent characteristics of the ocular media. Patients were also photographed with red free and infrared monochromatic imaging. The mean levels of autofluorescence were compared between patients without (group 1) and those with exudative AMD. Comparisons were made among patients with exudative AMD, examining the autofluorescence pattern in those without retinal vascular contribution to the exudative process (group 2) to those with retinal vascular contribution (group 3). MAIN OUTCOME MEASURES: Mean amounts and patterns of autofluorescence. RESULTS: A total of 54 patients was evaluated; 18 were in each group. The mean age was 75.4 years, and there was no difference in the mean ages among the groups (P = 0.16). There was no correlation of the autofluorescence measurements and the degree of nuclear sclerosis (P = 0.14). Patients with exudative AMD had more autofluorescence in the fellow eye than did eyes of patients without exudative AMD (P = 0.002). Patients in group 3 were more likely to have focal hyperpigmentation, particularly as imaged by infrared light (P = 0.015), and focal areas of intense autofluorescence (P = 0.001) than were patients in group 2. CONCLUSIONS: By use of this method of autofluorescence imaging, it was determined that the fellow eyes of patients with exudative AMD had larger amounts of autofluorescence than did the eyes of patients without a history of exudative AMD. Patients with retinal vascular anastomosis to the vascular proliferation of exudative AMD were much more likely to have focal areas of intense autofluorescence in their fellow eye that corresponded, for the most part, with focal areas of hyperpigmentation best seen by infrared monochromatic fundus photography. Because the amount of fluorescence is directly related to the amount of lipofuscin, which in turn is related to the cumulative amount of oxidative damage, these findings suggest possible explanations for certain patterns of vessel growth seen in exudative AMD. PMID- 12578787 TI - Agreement among ophthalmologists in evaluating fluorescein angiograms in patients with neovascular age-related macular degeneration for photodynamic therapy eligibility (FLAP-study). AB - OBJECTIVE: To determine intraobserver and interobserver variation for classifying types of choroidal neovascularizations (CNV) in exudative age-related macular degeneration (ARMD). DESIGN: Intraexaminer and interexaminer reliability study. PARTICIPANTS: Digital high-quality fluorescein angiograms of 40 patients with neovascular ARMD were evaluated independently by 16 retinal specialists. MAIN OUTCOME MEASURES: Fluorescein angiographies were presented in two randomized sequences (series A and B) to each masked reader for classification of type of CNV into classic, occult, or mixed with classic component of less or greater 50%. Agreement was evaluated by calculating kappa statistics (kappa) and intraclass correlation coefficients. RESULTS: The mean kappa coefficient was 0.64 +/- 0.11 for intraobserver variation, with a range from 0.44 to 0.89. For interobserver variation the intraclass correlation coefficients was calculated as 0.66 (95% confidence interval [CI] 0.56, 0.77) for series A and as 0.55 (95% CI 0.43, 0.67) for series B. CONCLUSIONS: Angiographic classification of CNV secondary to ARMD can vary considerably not only between observers but also for repeated evaluation by the same observer. Because various current and emerging treatments including photodynamic therapy are based on specific angiographic characteristics, accurate interpretation will become more important. PMID- 12578788 TI - Adjuvant indocyanine green in transpupillary thermotherapy for choroidal melanoma. AB - OBJECTIVE: To determine the effect of indocyanine green (ICG) injection on the regression pattern of choroidal melanomas treated with transpupillary thermotherapy (TTT). DESIGN: Prospective, randomized, controlled study. PARTICIPANTS: Of the 60 patients with posterior pole choroidal melanoma, 30 were managed with TTT alone (control group), and 30 received adjuvant ICG before TTT. METHODS: Subjects with selected choroidal melanoma were treated with TTT by using infrared radiation delivered from the diode laser. For those patients randomized to ICG administration, the start of TTT was based on the onset of fluorescence documented on ICG angiography. MAIN OUTCOME MEASURES: Final tumor thickness (1.5 mm) and temporal tumor thickness regression. RESULTS: The mean initial tumor basal diameter was 7.7 mm, and tumor thickness was 2.9 mm in the TTT treatment group. Tumor dimensions were initially 7.8 mm in the base and 3.1 mm in thickness in the TTT + ICG treatment group. During a median follow-up of 30 months (range, 6-49 months), the mean tumor thickness gradually decreased to 2.3 mm at month 3 and to 1.9 mm at month 6 after the initial session in the TTT treatment group and to 2.5 mm at month 3 and 2.2 mm at month 6 in the TTT + ICG treatment group. In the TTT treatment group, the reduction of tumor thickness was 19% at month 3 and 30% at month 6. In the TTT + ICG treatment group, the reduction of tumor thickness was 22% at month 3 and 31% at month 6. There were 12 tumors (40%) in the TTT treatment group and 15 (50%) in the TTT + ICG treatment group that measured 1.5 mm in thickness, with a flat ophthalmoscopic appearance. Tumor control was achieved in 55 tumors (92%). Two patients developed tumor recurrence in the TTT treatment group, and three developed recurrence in the TTT + ICG treatment group. A smaller initial tumor thickness, more TTT sessions, and a tumor location other than temporal were statistically predictive of a final tumor thickness of 1.5 mm. There was no apparent effect of ICG administration before TTT on the final tumor thickness and temporal tumor thickness regression. CONCLUSIONS: Despite the efficacy of TTT in the management of selected choroidal melanomas, adjuvant ICG administration before each TTT session does not seem to be beneficial in their regression pattern. Further research in vivo with a larger sample of patients and longer follow-up will be necessary to determine the role of ICG in TTT for choroidal melanomas. PMID- 12578790 TI - The histopathologic effects of transpupillary thermotherapy in human eyes. AB - OBJECTIVE: To determine the histopathologic effects of transpupillary thermotherapy (TTT) on the normal human retina. DESIGN: Prospective, noncomparative small case series. PARTICIPANTS: Three patients with eyes scheduled for enucleation because of the presence of a malignant intraocular tumor. INTERVENTION: Application of TTT to the posterior pole by using an 810-nm laser and the following laser parameters: 2-mm spot size, 60-second duration, and power settings of 430, 530, and 630 mW (low, medium, and high doses, respectively). Two or three TTT treatments at varying dose levels were performed in each eye. The eyes were then enucleated within 7 days of TTT, and light microscopy of serial sections was performed. MAIN OUTCOME MEASURES: Light microscopic histopathologic changes of the neurosensory retina, retinal pigment epithelium (RPE), and choroid in the areas of treatment compared with adjacent normal (control) tissue. Visual acuity, fundus appearance by slit-lamp biomicroscopy, and symptoms of pain, burning, or visual change were recorded before and after each TTT application. The degree of fundus pigmentation was also noted. RESULTS: Eight treatment spots from three eyes were analyzed. The first eye was judged to have a lightly pigmented fundus, and no histopathologic alterations were seen on light microscopy at any of the three dose levels. The second eye had a more pigmented fundus. This eye had minimal outer retinal changes in the area corresponding to the low-dose treatment, more prominent changes in the outer and middle layers in the medium-dose treatment area, and full-thickness retinal alterations, along with changes in the RPE and choroid, where the high dose was applied. The third eye was found to have an unexpected extension of pigmented choroidal melanoma under the fovea, and full-thickness retinal changes were observed in this area after a medium-dose application. No histopathologic changes were seen at the low dose in this eye. CONCLUSIONS: TTT applications resulted in a spectrum of histopathologic effects of the retina that are related to both energy level and fundus pigmentation. Mild or no changes were observed in most low- or medium-dose applications. More extensive retinal damage occurred with applications that used energy levels higher than what have been used in the clinical setting or when the fundus was more heavily pigmented. PMID- 12578791 TI - Expression of chemokine receptors, CXCR4 and CXCR5, and chemokines, BLC and SDF 1, in the eyes of patients with primary intraocular lymphoma. AB - OBJECTIVE: Chemokines have a range of biologic activities, including regulation of leukocyte trafficking, modulation of hematopoietic cell proliferation, and adhesion to extracellular matrix molecules. Specifically, B-lymphocyte chemoattractant (BLC); BCA-1; CXCL13, SCYB13) and stromal cell-derived factor-1 (SDF-1, CXCL12, SCYB12) are chemotactic for human B cells, and their ligands CXCR4 and CXCR5 are differentially expressed on B cells, including malignant B cells. We investigated the expression of these chemokine/chemokine receptors in eyes with primary intraocular B-cell lymphoma (PIOL). DESIGN: Observational case series (human tissue study). METHODS: Three freshly enucleated eyes with PIOL and a normal autopsied eye were frozen and sectioned. The sections were evaluated using immunohistochemistry (avidin-biotin-complex immunoperoxidase technique) for CXCR4, CXCR5, BLC, and SDF-1 to detect the expression and location. Reverse transcriptase-polymerase chain reaction was used to detect chemokine transcripts of CXCR4, CXCR5, BLC, and SDF-1 in PIOL and retinal pigment epithelium (RPE) cells after microdissection-either by laser capture (Arcturus) or by manual dissection-from frozen sections. MAIN OUTCOME MEASURES AND RESULTS: The three PIOL eyes showed similar pathology, with typical diffuse large B-lymphoma cells subjacent to the RPE. The eyes also demonstrated a similar chemokine profile. High expression levels of CXCR4 and CXCR5 were found limited to the lymphoma cells. In contrast, BLC protein was expressed in the RPE but not located in other ocular resident cells. SDF-1 was barely detected in a few RPE cells. CXCR4 and CXCR5 transcripts were detected abundantly in lymphoma cells, whereas BLC and SDF 1 transcripts were detected only in the RPE and not the malignant cells. No chemokine expression was detected on the RPE cells in the normal control eye. CONCLUSIONS: Chemokines and chemokine receptors selective for B cells were identified in RPE and malignant B cells, respectively. BLC, and possibly SDF-1, attracts both normal and malignant B-cells while promoting migration of only small numbers of T cells and macrophages. We propose that B-cell chemokines may be involved in the pathogenesis of PIOL by selectively attracting lymphoma cells to the RPE from the choroidal circulation. Our data suggest that inhibition of B cell chemoattractants could be a future strategy for the treatment of PIOL. PMID- 12578792 TI - Rhegmatogenous retinal detachment and uveitis. AB - PURPOSE: To evaluate the frequency, high-risk factors, and visual prognosis of rhegmatogenous retinal detachment (RRD) in patients with uveitis. DESIGN: Retrospective case-control study. PARTICIPANTS: We included 1387 consecutive patients with uveitis who consulted our uveitis clinic from January 1990 through December 1997 of whom 43 patients (46 eyes) with RRD were identified. The retinal detachment (RD) controls were 212 consecutive patients with RRD (221 eyes, first occurrence of RD, not associated with uveitis) who were admitted for surgery in the period from April 1999 to April 2000. The uveitis control group consisted of 150 age-matched patients (210 eyes) selected from the entire uveitis series. INTERVENTION: Retrospective analysis of clinical data. MAIN OUTCOME MEASURES: The presence of RRD and eventual risk factors for RRD, such as myopia, retinal lattice degeneration, prior intraocular surgery, anatomic location of uveitis, its specific diagnosis, and clinical manifestations. Furthermore, the surgical and nonsurgical outcomes of RRD, as well as the results of various treatment regimens, were analyzed. RESULTS: RRD was identified in 3.1% of the patients with uveitis. RRD was most frequently associated with panuveitis (6.6%). RRD was associated more frequently with infectious (7.6%) than noninfectious uveitis (2.1%). At the onset of RRD, uveitis was active in most (46%) affected eyes. Proliferative vitreoretinopathy was present in 30% of the uveitic RRD eyes at presentation in contrast to 12% of the RRD control eyes. In uveitic RRD, the retina was reattached in 59% of eyes with a single operation; the final anatomic reattachment rate was 88%. Finally, a visual acuity of less than 20/200 was present in 71% of the uveitic RRD eyes, 10% of which had no light perception. CONCLUSIONS: We discovered a high prevalence of RRD in patients with active panuveitis and infectious uveitis and document that uveitis in itself is a risk factor for the development of RRD. The visual prognosis of RRD in uveitis was poor because of the uveitis itself and the frequent development of proliferative vitreoretinopathy. PMID- 12578793 TI - Retinal microangiopathy in human immunodeficiency virus infection is related to higher human immunodeficiency virus-1 load in plasma. AB - PURPOSE: To evaluate the prevalence of retinal microangiopathy in human immunodeficiency virus (HIV)-1-infected patients and its association with virologic, immunologic, and sociodemographic parameters. DESIGN: Single-center cross-sectional study. PARTICIPANTS: One hundred eighty-eight HIV-1-positive individuals from a single outpatient clinic. METHODS: Human immunodeficiency virus-positive patients were screened for signs of HIV-associated retinal angiopathy. Plasma HIV-1 RNA and CD4-positive cell counts were monitored within 3 months of the ophthalmologic assessment. The absence or presence of angiopathy or of opportunistic viral retinitis was then correlated to data respecting CD4 positive cell count, plasma viral load of HIV-1, and sociodemographic parameters. MAIN OUTCOME MEASURES: Association between CD4-positive cell count, HIV-1 plasma viral load, sociodemographic parameters, and the manifestation of retinal microangiopathy. RESULTS: At the baseline consultation, 130 (69%) patients exhibited no retinal pathologic features, 45 (24%) manifested retinal angiopathy, and 13 (7%) had opportunistic viral retinitis. In univariate analysis, retinal angiopathy was associated with lower CD4-positive cell count and higher HIV-1 plasma viral load. In a multivariate logistic model, the presence of retinal microangiopathy was associated with higher age (P = 0.02) and higher viral load of HIV-1 (P < 0.005), but not with lower CD4 cell counts (P > 0.05). CONCLUSIONS: Human immunodeficiency virus-associated retinal microangiopathy is likely a multifactorial condition. Its presence is associated with higher age and higher replication of HIV-1 as measured by plasma HIV-1 RNA levels. In contrast to opportunistic infectious retinitis, the degree of immunodeficiency does not seem to be independently correlated with retinal angiopathy. PMID- 12578794 TI - Posterior segment complications in patients with hepatitis C treated with interferon and ribavirin. AB - PURPOSE: To determine the incidence and type of posterior segment complications associated with the use of interferon alpha 2b and ribavirin in patients with chronic hepatitis C. DESIGN: A prospective noncomparative case series. PARTICIPANTS: Forty-two patients (84 eyes). METHODS: Patients with chronic hepatitis C were evaluated for ocular changes while being treated with interferon alpha 2b and ribavirin. Patients were followed with sequential ocular examinations for 4 to 20 months. MAIN OUTCOME MEASURES: Occurrence of posterior segment complications while on interferon and ribavirin therapy. RESULTS: Twenty seven patients developed retinopathy. The retinopathy consisted of single-to multiple cotton-wool spots and retinal hemorrhage and was transient in all cases. An additional patient (age 46) presented with asymptomatic disc edema and hemorrhage. One other individual developed a symptomatic permanent monocular visual field defect. Therapy was discontinued in three patients because of severe posterior segment pathology. CONCLUSIONS: Our study demonstrated that a high incidence of retinopathy is associated with the treatment of hepatitis C using interferon and ribavirin, but that this form of retinopathy is relatively benign. Regular ophthalmic monitoring should be performed in patients undergoing this treatment. PMID- 12578795 TI - Proptosis after retrobulbar corticosteroid injections. AB - PURPOSE: This report describes the clinical, radiographic, and histopathologic findings in patients with proptosis secondary to lipomatosis after retrobulbar corticosteroid injection. DESIGN: Retrospective, noncomparative, interventional case series and review of the literature. METHODS: Five patients who developed symptomatic unilateral proptosis after steroid injection were studied and the literature was reviewed. RESULTS: No previous description was found in the literature. All five cases were studied with computed tomography, and two cases were confirmed with histopathology. No fibrosis or granulomatous inflammation was identified. CONCLUSIONS: Orbital lipomatosis is a potential complication of retrobulbar steroid injections. Symptomatic relief can be provided by a transconjunctival approach to the lower lid fat compartment. PMID- 12578796 TI - On the failure to disclose sibling manuscripts. PMID- 12578797 TI - Partner characteristics, intensity of the intercourse, and semen exposure during use of the female condom. AB - The objective of this study was to assess how characteristics of the intercourse and the couple relate to semen exposure during use of the female condom. From 1996 to 1998, 210 women in Birmingham, Alabama, were trained to use the female condom and follow study procedures during a group session and individually practiced inserting the device. The outcome was semen exposure as defined by comparing pre- and postcoital prostate-specific antigen levels in vaginal fluid. Women who had high income levels had lower rates of semen exposure (odds ratio (OR) = 0.3, 95% confidence interval (CI): 0.2, 0.7), while those in a relationship of less than 2 years were at greater risk (OR = 2.4, 95% CI: 1.3, 4.1). Couples with a large disparity in vaginal fundus size and penis size were at increased risk of semen exposure (OR = 2.7, 95% CI: 1.2, 6.0). Engaging in very active intercourse also increased the risk (OR = 1.7, 95% CI: 1.1, 2.6). Thus, the protective effect of the female condom appears to be a function of user and intercourse-specific characteristics. Future studies of male condom efficacy should focus on collecting detailed data about the users and characteristics of intercourse to predict failure accurately. PMID- 12578798 TI - Efficacy of the female condom as a barrier to semen during intercourse. AB - In 1996-1998, the authors measured prostate-specific antigen (PSA) in vaginal fluid to assess the frequency of female condom failure and to evaluate the association of self-reported failure with semen exposure. Women at low risk of sexually transmitted diseases (n = 210) were recruited in Birmingham, Alabama. They were trained to use the female condom, sample vaginal fluid before and after condom use, and complete forms to report problems during each use. Semen exposure was assessed by comparing pre- and postcoital PSA levels in vaginal fluid. A total of 175 women used 2,232 condoms. The rate of semen exposure ranged from 7% to 21% of condom uses, depending on the exposure criterion. Exposure was more likely (21-34%) and more intense (mean postcoital PSA, 24.7 ng/ml) if participants reported a mechanical problem versus other problems or no problems (exposure rate, 5-20% in both instances; mean postcoital PSA, 9.6 and 7.8 ng/ml, respectively). In logistic regression analyses for repeated measurements, user reported problems accounted for less than 59% of the instances of semen exposure. The female condom prevented semen exposure in 79-93% of condom uses. Exposure was associated with user-reported problems but also occurred in their absence. Reported problems and semen exposure decreased with user experience. PMID- 12578799 TI - Invited commentary: condom effectiveness-will prostate-specific antigen shed new light on this perplexing problem? PMID- 12578801 TI - Genetic and environmental contributions to cardiovascular disease risk in American Indians: the strong heart family study. AB - The aims of the Strong Heart Family Study are to clarify the genetic determinants of cardiovascular disease (CVD) risk in American Indians and to map and identify genes for CVD susceptibility. The authors describe the design of the Strong Heart Family Study (conducted between 1998 and 1999) and evaluate the heritabilities of CVD risk factors in American Indians from this study. In the first phase of the study, approximately 950 individuals, aged 18 years or more, in 32 extended families, were examined. The examination consisted of a personal interview, physical examination, laboratory tests, and an ultrasound examination of the carotid arteries. The phenotypes measured during the physical examination included anthropometry, lipoproteins, blood pressure, glycemic status, and clotting factors. Heritabilities for CVD risk factor phenotypes were estimated using a variance component approach and the program SOLAR. After accounting for the effects of covariates, the authors detected significant heritabilities for many CVD risk factor phenotypes (e.g., high density lipoprotein cholesterol (heritability = 0.50) and diastolic blood pressure (heritability = 0.34)). These results suggest that heredity explains a substantial proportion of the variability of CVD risk factors and that these heritabilities are large enough to warrant a search for major risk factor genes. PMID- 12578802 TI - Area characteristics, individual-level socioeconomic indicators, and smoking in young adults: the coronary artery disease risk development in young adults study. AB - The 10-year follow-up examination in 1995-1996 to the population-based Coronary Artery Disease Risk Development in Young Adults Study was used to compare the strength with which socioeconomic indicators at the individual and area levels are related to smoking prevalence and to investigate contextual effects of area characteristics. When categories based on similar percentile cutoffs were compared, differences across area categories in the odds of smoking were smaller than differences across categories based on individual-level indicators. In Whites, there was evidence of a significant contextual effect of area characteristics on smoking: Living in the most disadvantaged area quartiles was associated with 50-110% higher odds of smoking, even after controlling for individual-level socioeconomic indicators. Clear contextual effects of area characteristics were not present in Blacks, but there was evidence that contextual effects may emerge at higher levels of individual-level socioeconomic position. Similar results were obtained for census tracts and block groups. Even in the presence of contextual effects, area measures may underestimate associations of individual-level variables with health outcomes. On the other hand, as illustrated by the presence of contextual effects, area- and individual level measures are likely to tap into different constructs. PMID- 12578803 TI - Relation between cognitive function and mortality in middle-aged adults: the atherosclerosis risk in communities study. AB - An independent, inverse association between cognitive function and all-cause mortality has been reported in elderly cohorts. The purpose of this study was to determine whether the same association exists in middle-aged persons. The Atherosclerosis Risk in Communities Study is a cohort study initiated in 1987 to investigate the development of atherosclerosis in middle-aged persons. Three cognitive function measures were included in the second cohort examination conducted from 1990 to 1992 when the participants were aged 48-67 years: the Delayed Word Recall Test (DWRT), the Digit Symbol Substitution Test (DSST) (a subtest from the Wechsler Adult Intelligence Scale-Revised), and the Word Fluency Test from the Multilingual Aphasia Examination. Cox proportional hazards modeling was used to determine whether all-cause mortality ascertained through 1997 was associated with each measure after adjustment for sociodemographic, biologic, psychologic, and behavioral risk factors. Without adjustment, there was a significantly lower mortality hazard associated with higher scores on all three measures. After covariate adjustment, the hazard ratios for the DWRT and the DSST remained significant (hazard ratio1-point DWRT score increment = 0.90, 95% confidence interval: 0.84, 0.97; hazard ratio 7-point DSST score increment = 0.86, 95% confidence interval: 0.80, 0.93). Cognitive function measured in middle age appears to have prognostic importance for life expectancy similar to that reported in elderly adults. PMID- 12578804 TI - Prospective study of antioxidant micronutrients in the blood and the risk of developing prostate cancer. AB - Antioxidant micronutrients may have chemopreventive effects. The authors examined the associations between prediagnostic blood levels of micronutrients and prostate cancer risk in two nested case-control studies of 9,804 and 10,456 male residents of Washington County, Maryland, who donated blood in 1974 (CLUE I) and 1989 (CLUE II), respectively. Until 1996, 182 men for whom adequate serum remained for assays in the CLUE I cohort and 142 men in the CLUE II cohort developed prostate cancer. Each case was matched with two controls by age, gender, race, and date of blood donation. In both cohorts, cases and controls had similar concentrations of alpha-carotene, beta-carotene, total carotene, beta cryptoxanthin, lutein, lycopene, retinol, and ascorbic acid; serum alpha tocopherol was weakly associated with prostate cancer risk. Higher retinyl palmitate concentrations were associated with a lower risk in CLUE I but not CLUE II. In CLUE I, cases had lower concentrations of gamma-tocopherol than did controls (p = 0.02), but no dose-response trend was observed. A strong inverse association between gamma-tocopherol and prostate cancer risk was observed in CLUE II. Findings do not replicate previous reports of a protective association between lycopene and prostate cancer, but they suggest potential chemopreventive effects of gamma-tocopherol on prostate cancer. PMID- 12578805 TI - Antioxidant micronutrients and risk of rheumatoid arthritis in a cohort of older women. AB - The association of antioxidant vitamins and trace elements from foods and supplements with risk of rheumatoid arthritis was evaluated in a prospective cohort study of 29,368 women who were aged 55-69 years at baseline in 1986. Through 1997, 152 cases of rheumatoid arthritis were identified. After controlling for other risk factors, greater intakes (highest tertile vs. lowest) of supplemental vitamin C (relative risk (RR) = 0.70, 95% confidence interval (CI): 0.48, 1.09; p-trend = 0.08) and supplemental vitamin E (RR = 0.72, 95% CI: 0.47, 1.12; p-trend = 0.06) were inversely associated with rheumatoid arthritis. There was no association with total carotenoids, alpha- or beta-carotene, lycopene, or lutein/zeaxanthin, while there was an inverse association with beta cryptoxanthin (RR = 0.59, 95% CI: 0.39, 0.90; p-trend = 0.01). Greater use of supplemental zinc (RR = 0.39, 95% CI: 0.17, 0.88; p-trend = 0.03) was inversely associated with rheumatoid arthritis, while any use of supplemental copper (RR = 0.54, 95% CI: 0.28, 1.03) and manganese (RR = 0.50, 95% CI: 0.23, 1.07) showed suggestive inverse associations with rheumatoid arthritis. Greater intakes of fruit (RR = 0.72, 95% CI: 0.46, 1.12; p-trend = 0.13) and cruciferous vegetables (RR = 0.65, 95% CI: 0.42, 1.01; p-trend = 0.07) also exhibited trends toward inverse associations with risk. When the antioxidants were modeled together, only beta-cryptoxanthin and supplemental zinc were statistically significant predictors. Intake of certain antioxidant micronutrients, particularly beta cryptoxanthin and supplemental zinc, and possibly diets high in fruits and cruciferous vegetables, may be protective against the development of rheumatoid arthritis. PMID- 12578806 TI - Effects of exposure measurement error when an exposure variable is constrained by a lower limit. AB - Epidemiologic studies routinely suffer from bias due to exposure measurement error. In this paper, the authors examine the effect of measurement error when the exposure variable of interest is constrained by a lower limit. This is an important consideration, since often in epidemiologic studies an exposure variable is constrained by a lower limit such as zero or a nonzero detection limit. In this paper, attenuation of exposure-disease associations is defined within the framework of a classical model of uncorrelated additive error. Then, the special case of nonlinearity due to the effect of a lower threshold is examined. A general model is developed to characterize the effect of random measurement error when there is a lower threshold for recorded values. Findings are illustrated under the assumption that the true exposure follows the lognormal and gamma distributions. The authors show that the direction and magnitude of bias in estimated exposure-response associations depends on the population distribution of the exposure, the magnitude of the recording threshold, the value assigned to below-threshold measurement results, and the variance in the measured exposure due to random measurement error. PMID- 12578807 TI - Statistical analysis of correlated data using generalized estimating equations: an orientation. AB - The method of generalized estimating equations (GEE) is often used to analyze longitudinal and other correlated response data, particularly if responses are binary. However, few descriptions of the method are accessible to epidemiologists. In this paper, the authors use small worked examples and one real data set, involving both binary and quantitative response data, to help end users appreciate the essence of the method. The examples are simple enough to see the behind-the-scenes calculations and the essential role of weighted observations, and they allow nonstatisticians to imagine the calculations involved when the GEE method is applied to more complex multivariate data. PMID- 12578809 TI - Re: "Risk of hepatitis C virus among young adult injection drug users who share injection equipment". PMID- 12578810 TI - Implications of intracrine hormone action for physiology and medicine. PMID- 12578811 TI - Prostaglandin modulation of venoconstriction to physiological stress in normals and heart failure patients. AB - Prostaglandins released from blood vessels modulate vascular tone, and inhibition of their production during exogenous infusions of catecholamines causes increased venoconstriction. To determine the influence of prostaglandin production on venoconstriction during physiological stimuli known to cause sympathetic activation, and to assess its importance in chronic heart failure (CHF), we studied 11 normal subjects (62 +/- 4 yr) and 14 patients with CHF (64 +/- 2 yr, left ventricular ejection fraction 23 +/- 1%, New York Heart Association classes II and III) (means +/- SE). Dorsal hand vein distension was measured during mental arithmetic (MA), cold pressor test (CPT), and lower body negative pressure (LBNP; -10 and -40 mmHg), with saline infusion in one hand and local indomethacin (cyclooxygenase inhibitor) infusion (3 microg/min) in the other. Acetylcholine (0.01-1 nmol/min) dilated veins preconstricted with PGF(2alpha) in normals but, consistent with endothelial dysfunction, barely did so in CHF patients (P = 0.001). Nonendothelial venodilation to sodium nitroprusside (0.3-10 nmol/min) was not different between normals and CHF patients. Resting venous norepinephrine levels were higher in CHF patients (2,812 +/- 420 pmol/l) than normals (1,418 +/- 145 pmol/l, P = 0.007). In normals, indomethacin caused increased venoconstriction to MA (from 4.9 +/- 1.5 to 19.2 +/- 4.5%, P = 0.022) and CPT (from 2.9 +/- 3.8 to 17.6 +/- 4.2%, P = 0.007). In CHF, indomethacin caused increased venoconstriction to MA (from 6.6 +/- 3.9% to 19.0 +/- 4.5%, P = 0.014), CPT (from 9.6 +/- 2.1% to 20.1 +/- 3.7%, P = 0.001), and -40 mmHg LBNP (from 10.7 +/- 3.0% to 23.2 +/- 3.8%, P = 0.041). Control responses for all tests were not different between normals and CHF patients. The effects of indomethacin on venoconstriction to MA and CPT were not different between normals and CHF patients, but venoconstriction to -40 mmHg LBNP was accentuated in CHF patients (P = 0.036). Inhibition of prostaglandins by indomethacin significantly enhances hand vein constriction to physiological stimuli in both normals and CHF patients, although a differential effect exists for LBNP. PMID- 12578812 TI - Effective protection by NHE-1 inhibition in ischemic and reperfused heart under preconditioning blockade. AB - We compared the protective effects of ischemic preconditioning (IPC) and the Na(+)/H(+) exchanger-1 (NHE-1) inhibitor cariporide in isolated rat hearts subjected to global ischemia (45 or 90 min) and 30-min reperfusion and determined the protective effects of cariporide under IPC blockade with the mitochondrial ATP-sensitive K(+) channel blocker 5-hydroxydecanoate (5-HD). With 45-min ischemia, both IPC and cariporide equally increased maximum recovery of left ventricular developed pressure twofold (P < 0.05), although recovery was significantly greater with cariporide for the first 15 min of reperfusion. 5-HD almost completely blocked the protective effects of IPC on recovery but had no influence on the salutary effects of cariporide. With 90-min ischemic control, recovery was only 3% of preischemia and was unaffected by IPC, although cariporide increased recovery to approximately 30% (P < 0.05). This was associated with a 37% preservation of viable cardiac cells, whereas no structurally intact cells were found in either IPC or control hearts. Our study shows that NHE-1 inhibition is a more effective cardioprotective strategy than IPC in this model, possibly because of enhanced myocyte salvage, and because protection by NHE-1 inhibition is completely unaffected by IPC blockade with 5 HD. PMID- 12578813 TI - I kappa B overexpression in cardiomyocytes prevents NF-kappa B translocation and provides cardioprotection in trauma. AB - This study examined the effects of either IkappaBalpha overexpression (transgenic mice) or N-acetyl-leucinyl-leucinyl-norleucinal (ALLN) administration (proteosome inhibitor in wild-type mice) on cardiomyocyte secretion of tumor necrosis factor alpha (TNF-alpha) and on cardiac performance after burn trauma. Transgenic mice were divided into four experimental groups. IkappaBalpha overexpressing mice were given a third-degree scald burn over 40% of the total body surface area or wild type littermates were given either a scald or sham burn to provide appropriate controls. Pharmacological studies included ALLN (20 mg/kg) administration in either burned wild-type mice or wild-type shams. Burn trauma in wild-type mice promoted nuclear factor-kappaB (NF-kappaB) nuclear translocation, cardiomyocyte secretion of TNF-alpha, and impaired cardiac performance. IkappaBalpha overexpression or ALLN treatment of burn trauma prevented NF-kappaB activation in cardiac tissue, prevented cardiomyocyte secretion of TNF-alpha, and ablated burn mediated cardiac contractile dysfunction. These data suggest that NF-kappaB activation and inflammatory cytokine secretion play a significant role in postburn myocardial abnormalities. PMID- 12578814 TI - Effects of hypoxia on heme oxygenase expression in human chorionic villi explants and immortalized trophoblast cells. AB - Although hypoxia induces heme oxygenase (HO)-1 protein and mRNA expression in many cell types, hypoxia has also been shown to decrease HO-1 mRNA and protein expression. We tested the hypothesis that 24-h preexposure to hypoxia in human placental preparations suppresses HO protein expression and enzymatic function. Immortalized HTR-8/SVneo first-trimester trophoblast cells and explants of normal human chorionic villi (CV) from term placentas were cultured for 24 h in 1%, 5%, or 20% O(2). HO protein levels were determined by Western blot analysis, and microsomal HO activity was measured. HO-2 protein content was decreased by 17% and 5% in human trophoblast cells after 24-h exposure to 1% and 5% O(2), respectively, versus 20% O(2). In contrast, HO-2 protein content in CV explants was unaffected by changes in oxygenation. HO-1 protein content, which was barely detectable in both biological systems, was not affected by changes in oxygenation. Similarly, HO enzymatic activity was unchanged in both preparations after 24-h exposure to 1%, 5%, or 20% O(2). The above data do not support the hypothesis that hypoxia in the human placenta suppresses both HO protein content and HO protein function. The present observations reinforce the necessity to determine both HO protein expression and function. PMID- 12578815 TI - A1 adenosine receptor overexpression attenuates ischemia-reperfusion-induced apoptosis and caspase 3 activity. AB - We tested the hypothesis that myocardial ischemia-reperfusion (I/R)-induced apoptosis is attenuated in transgenic mice overexpressing cardiac A(1) adenosine receptors. Isolated hearts from transgenic (TG, n = 19) and wild-type (WT, n = 22) mice underwent 30 min of ischemia and 2 h of reperfusion, with evaluation of apoptosis, caspase 3 activity, function, and necrosis. I/R-induced apoptosis was attenuated in TG hearts. TG hearts had less I/R-induced apoptotic nuclei (0.88 +/ 0.10% vs. 4.22 +/- 0.24% terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells in WT, P < 0.05), less DNA fragmentation (3.30 +/- 0.38-fold vs. 4.90 +/- 0.39-fold over control in WT, P < 0.05), and less I/R induced caspase 3 activity (145 +/- 25% over nonischemic control vs. 234 +/- 31% in WT, P < 0.05). TG hearts also had improved recovery of function and less necrosis than WT hearts. In TG hearts pretreated with LY-294002 (3 microM) to evaluate the role of phosphosinositol-3-kinase in acute signaling, there was no change in the functional protection or apoptotic response to I/R. These data suggest that cardioprotection with transgenic overexpression of A(1) adenosine receptors involves attenuation of I/R-induced apoptosis that does not involve acute signaling through phosphoinositol-3-kinase. PMID- 12578816 TI - Activity-dependent role of NMDA receptors in transmission of cardiac mechanoreceptor input to the NTS. AB - Evidence suggests that transmission of barosensitive input from arterial baroreceptors and cardiac mechanoreceptors at nucleus tractus solitarius (NTS) neurons involves non-N-methyl-d-aspartate (NMDA) glutamate receptors, but there is a possibility that the contribution of NMDA receptors might increase during periods of increased afferent input, when enhanced neuronal depolarization could increase the activation of NMDA receptors by removal of a Mg(2+) block. Thus the effects of NMDA on cardiac mechanoreceptor-modulated NTS neuronal discharges were examined at different levels of arterial pressure used to change cardiac mechanoreceptor afferent input. To determine whether the response was specific to NMDA, (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid (AMPA) was also administered at different levels of neuronal discharge. In anesthetized dogs, neuronal activity was recorded from the NTS while NMDA or AMPA was picoejected at high versus low arterial stimulating pressures. NMDA, but not AMPA, produced a significantly greater discharge of mechanoreceptor-driven NTS neurons at higher versus lower levels of stimulating pressure. These data suggest that the role played by NMDA receptors is greater during periods of enhanced neuronal depolarization, which could be produced by increases in afferent barosensitive input. PMID- 12578817 TI - Effect of increased expression of cytoskeletal protein vinculin on ischemia reperfusion injury in ventricular myocytes. AB - The transition from reversible to irreversible ischemic injury (ischemia reperfusion, I/R) occurs coincident with the loss of vinculin, a cytoskeletal protein involved in the attachment of the myofibrils to the sarcolemmal membrane. If the loss of vinculin were critical to the development of I/R, then increased levels of vinculin would be predicted to delay the onset of irreversible injury assuming that the protein is functional and localized to the proper subcellular site. The present study determined whether increased expression of vinculin, specifically in the cytoskeletal compartment, would provide protection from I/R injury. Neonatal rat myocytes were cultured and infected with a newly created replication-deficient adenovirus driving the expression of vinculin. I/R was induced with chemical inhibitors of glycolysis and mitochondrial respiration. Irreversible cell injury was assessed with lactate dehydrogenase (LDH) release. Virus-infected myocytes expressed significantly more vinculin in the cytoskeletal fraction and increased the expression of paxillin but sustained the same amount of injury in response to simulated I/R as control cells (n = 4; P = not significant, paired t-test). Hypothermic I/R (ischemia at 25 degrees C) resulted in a significant reduction in LDH release (P 98%) and repaired HSE-induced mitochondrial damages within 48 h. However, iB16M cells with low mtGSH levels were highly susceptible to TNF-alpha induced oxidative stress and death. Therefore depletion of mtGSH levels may represent a critical target to challenge survival of invasive cancer cells. PMID- 12578842 TI - Thermodynamic analysis of the increased stability of major histocompatibility complex class II molecule I-Ek complexed with an antigenic peptide at an acidic pH. AB - The differential scanning calorimetry analysis of the murine major histocompatibility complex class II molecule, I-E(k), in complex with an antigenic peptide derived from mouse hemoglobin, showed that the thermal stability at the mildly acidic pH is higher than that at the neutral pH. Although the thermal unfolding of I-E(k)-hemoglobin was irreversible, we extracted the equilibrium thermodynamic parameters from the kinetically controlled heat capacity curves. Both the denaturation temperatures and the enthalpy changes were almost independent of the heating rate over 1 degrees C per min. The linear relation between the denaturation temperature and the calorimetric enthalpy change provided the heat capacity changes, which are classified into one for the mildly acidic pH region and another for the neutral pH region. The equilibrium thermodynamic parameters showed that the increased stability at the mildly acidic pH is because of the entropic effect. These thermodynamic data provided new insight into the current structural model of a transition to an open conformation at the mildly acidic pH, which is critical for the peptide exchange function of major histocompatibility complex class II in the endosome. PMID- 12578843 TI - Crucial role of TCL/TC10beta L, a subfamily of Rho GTPase, in adipocyte differentiation. AB - The events at the beginning of adipocyte differentiation are not well known. We previously cloned the genes expressed early in the differentiation of mouse 3T3 L1 preadipocyte cells. One of them, similar in sequence to human TC10, was identified as TC10-like/TC10betaLong (TCL/TC10betaL), a new Rho GTPase by the cloning of full-length cDNA. The expression of TCL/TC10betaL increased rapidly right after the addition of inducers for differentiation, whereas the levels of other Rho family genes were unchanged at this stage. The antisense TCL/TC10betaL expressing experiment revealed that the differentiation of 3T3-L1 cells into adipocytes was inhibited. Moreover, the sense TCL/TC10betaL-expressing experiment using NIH-3T3 cells, which do not usually differentiate into adipocytes, clearly showed the accumulation of oil droplets as well as the elevated expression of various adipogenic marker genes in the presence of the ligand for peroxisome proliferator-activated receptor gamma (PPARgamma). These results strongly indicated that TCL/TC10betaL has a crucial role in the early stage of adipocyte differentiation, probably linked to the PPARgamma pathway. Using a subtraction protocol, the genes specifically regulated by TCL/TC10betaL were also isolated. The expression pattern of some of them was similar to TCL/TC10betaL expression in adipogenesis, suggesting that the expression of these genes would be regulated by TCL/TC10betaL. PMID- 12578845 TI - Tracking dendritic cells: use of an in situ method to label all blood leukocytes. AB - Here we describe an in situ procedure with a labeling index (percent of labeled blood leukocytes) >98%, which is high enough to permit the direct tracking of dendritic cell (DC) precursors from blood into lymphoid tissues, while circumventing the pitfalls associated with in vitro labeling. DC and lymphocytes have similar blood to afferent lymph migratory capabilities. This method has additional applications in tracking other rare cell populations in both normal and pathological states. PMID- 12578844 TI - Lack of oral tolerance in aging is due to sequential loss of Peyer's patch cell interactions. AB - Our past studies showed that Peyer's patches were required for the induction of oral tolerance to the protein antigen ovalbumin (OVA), but not to the hapten 2,4,6-trinitrobenzene sulfonic acid (TNBS). In the present study, the effects of immunosenescence on oral tolerance induction were assessed with these two toleragens. Significant reductions in OVA-specific serum IgG antibody and CD4(+) T cell responses to subsequent challenge were observed in OVA-fed, young adult mice. Importantly, these reduced anti-OVA antibody responses were associated with delayed-type hypersensitivity, and antigen-induced CD4(+) T(h)1- and T(h)2-type cytokine responses. On the other hand, aged mice fed OVA failed to develop oral tolerance. Thus, CD4(+) T cells from Peyer's patches produced selected T(h)2- but no T(h)1-type cytokines. The TNP-specific serum IgG antibody and T cell responses were significantly diminished by prior TNBS feeding in young adult, 6- to 8-month old and 12- to 14-month-old, but not in senescent, 2-year-old mice. Finally, we have directly assessed dendritic cell subsets and T cell responses in Peyer's patches, and their function in tolerance induction was impaired at an earlier stage of life. These results suggest that lack of oral tolerance to the protein OVA during aging is the result of dysfunctional Peyer's patches. PMID- 12578846 TI - CD80 and CD86 C domains play an important role in receptor binding and co stimulatory properties. AB - CD80 and CD86 expressed on the surface of antigen-presenting cells interact with cytotoxic T lymphocyte antigen-4 [CTLA-4 (CD152)] expressed on activated T cells and mediate critical T cell inhibitory signals. CD80 and CD86 are type I glycoproteins, and are made up of two extracellular (EC) Ig-like domains-a transmembrane region and a cytoplasmic tail. The N-terminal (V domain) and membrane-proximal (C) domains share homology with the variable region (V) and the constant region (C) of Ig respectively. Co-crystallographic structures of both CD80 and CD86 bound to CTLA-4 indicate that there is no direct interaction of the C domain of either CD80 or CD86 with the CTLA-4. In contrast, previous mutagenesis studies have identified specific amino acids within the C domain of CD80 that are critical for CTLA-4 binding. To further understand the importance of C domains in the functioning of CD80 and CD86, we constructed chimeric human CD80 and CD86 molecules by swapping their respective C domains, and tested their ability to stimulate T cells. A Chinese hamster ovary (CHO) cell line expressing CD86 activated murine T cells. In contrast, CHO cells expressing either CD80 or a chimeric construct of the CD86 V domain and the CD80 C domain showed a significantly reduced activation. Our studies further demonstrated that the decreased activation by cells expressing the CD80 or a chimera containing CD80 C domain is most likely due to enhanced CTLA-4 binding. From these results we conclude that C domains play a critical, albeit indirect, role in determining CTLA-4 binding affinities and co-stimulatory properties. PMID- 12578847 TI - SIGN-R1, a novel C-type lectin expressed by marginal zone macrophages in spleen, mediates uptake of the polysaccharide dextran. AB - The marginal zone macrophages of the spleen are implicated in the clearance of polysaccharides, but underlying mechanisms need to be pinpointed. SIGN-R1 is one of five recently identified mouse genes that are homologous to human DC-SIGN and encode a single, external, C-terminal C-type lectin domain. We find that a polyclonal antibody to a specific SIGN-R1 peptide reacts primarily and strongly with a subset of macrophages in the marginal zone of spleen and lymph node medulla. In both sites, SIGN-R1 exists primarily in an aggregated form, resistant to dissociation into monomers upon boiling in SDS under reducing conditions. Upon transfection into three different cell lines, high-mol.-wt forms bearing SIGN-R1 are expressed, as well as reactivity with ER-TR9, a mAb previously described to react selectively with marginal zone macrophages. SIGN-R1-expressing macrophages preferentially sequester dextrans following i.v. injection. Likewise, when phagocytic cells are enriched from spleen and tested in culture, dextran is selectively endocytosed by a subset of very large SIGN-R1(+) cells representing approximately 5% of total released macrophages. Uptake of FITC-dextran by these macrophages in vivo and in vitro is blocked by ER-TR9 and polyclonal anti-SIGN-R1 antibodies. Following transfection with SIGN-R1, cell lines become competent to endocytose dextrans. The dextran localizes primarily to compartments lacking transferrin receptor and the LAMP-1 CD107a panlysosomal antigen. Therefore, SIGN R1 mediates the uptake of dextran polysaccharides, and it is predominantly expressed in the macrophages of the splenic marginal zone and lymph node medulla. PMID- 12578848 TI - Evidence to support the cellular mechanism involved in serum IgG homeostasis in humans. AB - IgG is the most abundant serum antibody and is an essential component of the humoral immune response. It is known that the 'neonatal' Fc receptor (FcRn) plays a role in maintaining constant serum IgG levels by acting as a protective receptor which binds and salvages IgG from degradation. However, the cellular mechanism that is involved in serum IgG homeostasis is poorly understood. In the current study we address this issue by analyzing the intracellular fate in human endothelial cells of IgG molecules which bind with different affinities to FcRn. The studies show that IgG which do not bind to FcRn accumulate in the lysosomal pathway, providing a cellular explanation for short serum persistence of such antibodies. We have also investigated the saturability of the homeostatic system and find that it has limited capacity. Our observations have direct relevance to the understanding and treatment of IgG deficiency, and to the effective application of therapeutic antibodies. PMID- 12578849 TI - In vitro evidence for participation of DEC-205 expressed by thymic cortical epithelial cells in clearance of apoptotic thymocytes. AB - Binding of apoptotic cells was compared after incubation of thymocytes with two clones of murine thymic stromal cells to which CD4(+)/CD8(+) thymocytes attach. With the BA/10, but not the BA/2, clone, thymocytes with apoptotic morphology were bound irreversibly. These tightly bound thymocytes were further identified as apoptotic in terms of active caspase-3 and DNA fragmentation assayed in situ. FACS analysis indicated that the apoptotic thymocytes are at an early double positive stage and results with mice mutant for the Fas gene showed that the Fas Fas ligand system is not involved. Comparison of BA/10 and BA/2 cells showed that the former, but not the latter, can be induced to express CDR-1 antigen which is characteristic of cortical epithelial thymic stroma and constitutively express DEC-205, a surface protein common to cortical thymic epithelium and dendritic cells. Antibody NLDC-145 that is specific for the DEC-205 protein strongly reduced the number of stromal cells with bound apoptotic thymocytes. Preincubation of thymocytes in dexamethasone dramatically increased the number of bound apoptotic cells, indicating that the thymic cortical epithelial cells can participate in clearance of apoptotic thymocytes through involvement of DEC-205. PMID- 12578850 TI - Peptides based on the complementarity-determining regions of a pathogenic autoantibody mitigate lupus manifestations of (NZB x NZW)F1 mice via active suppression. AB - Two peptides based on the complementarity-determining regions (CDR) 1 and 3 (pCDR1 and pCDR3) of a murine monoclonal anti-DNA autoantibody that expresses the common idiotype 16/6Id were shown to down-regulate systemic lupus erythematosus (SLE)-associated T cell responses and to prevent the development of clinical symptoms in the SLE-prone mice, (NZB x NZW)F(1). In the present study the ability of the CDR-based peptides to treat an already established disease was tested. Mice were given 10 weekly injections of peptides either i.v. or s.c. The treatment led to a moderate reduction in the anti-DNA autoantibody titer, and a significant decrease in proteinuria and kidney pathology. The CDR-based peptides affected the pathogenic isotypes (IgG2a and IgG3) of the anti-DNA antibodies in the serum and in immune complexes in the kidneys. Both peptides mitigated disease manifestations and prolonged the survival of mice that were treated starting at the age of 7 months when full-blown disease was already developed. Furthermore, some beneficial effects of treatment with the CDR-based peptides could be adoptively transferred to diseased recipients. A reduction in the secretion of IL 2, IFN-gamma, IL-4 and IL-10 was detected in supernatants of splenocytes of the treated mice. In contrast, treatment up-regulated the immunosuppresive cytokine transforming growth factor-beta. Thus the ameliorating effect of the CDR-based peptides on SLE manifestations is at least partially via the immunomodulation of the cytokine profile. PMID- 12578851 TI - Essential role for Vav1 in activation, but not development, of gammadelta T cells. AB - Vav1 is a guanine nucleotide exchange factor essential in the development and function of alphabeta lineage T cells. Here we report that in contrast to profound effects on pre-TCR- or alphabeta TCR-dependent events in thymocyte development, Vav1 deficiency has no detectable effect on the development of gammadelta T cells. Strikingly, however, these gammadelta T cells are markedly deficient in signaling through the gammadelta TCR, as evidenced by a lack of proliferation and cytokine production in response to stimulation with anti gammadelta TCR antibodies. We propose that Vav1 has a unique and non-redundant role in the initiation of signaling downstream of the gammadelta TCR in lymphocytes. PMID- 12578852 TI - Synergy between CpG- or non-CpG DNA and specific antigen for B cell activation. AB - DNA or oligodeoxynucleotides (ODN) containing unmethylated CpG motifs (CpG DNA) activate antigen-presenting cells and switch on T(h)1 immunity to antigen. B cells are synergistically activated by CpG DNA in combination with non physiologic B cell stimulators such as polyclonal mitogen and surface Ig cross linkers. This study shows the unexpected finding that not only CpG ODN, but also non-CpG and methylated ODN synergize with specific antigen, hen egg lysozyme (HEL), in stimulating HEL-specific B cells to proliferate, to express the early activation marker CD69 and to activate the NF-kappaB pathway. In vivo, non-CpG and methylated CpG ODN also enhanced anti-HEL antibody production in HEL immunized mice, with a bias towards the production of T(h)1-associated isotypes. The synergy with all ODN to enhance B cell immune function was epitope-specific since neither denatured HEL nor other antigens enhanced the ODN effect on HEL specific B cells. Furthermore, the synergy was independent of whether the ODN backbone was phosphorothioate or phosphodiester, or whether natural vertebrate genomic DNA was used. In all functional analyses, non-CpG and methylated CpG ODN showed lower activity than CpG ODN. These studies demonstrate that the presence of specific physiologic antigen might broaden the spectrum of DNA/ODN that stimulate B cells, with potential implications for the initiation and regulation of normal and pathologic immune responses. PMID- 12578853 TI - Differential effect of serotonin on cytokine production in lipopolysaccharide stimulated human peripheral blood mononuclear cells: involvement of 5 hydroxytryptamine2A receptors. AB - In order to provide additional insight into the in vivo significance of serotonin [5-hydroxytryptamine (5-HT)] in inflammation, we examined its effect on the production of tumor necrosis factor (TNF)-alpha, IL-1alpha, IL-1beta, IL-6, IL-10 and IL-1 receptor antagonist in lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMC). 5-HT inhibited TNF-alpha production and increased IL-1beta production in PBMC. The level of IL-1beta-converting enzyme/caspase-1 remained unchanged, suggesting that the effect of 5-HT is not directly related to the IL-1beta maturation process. TNF-alpha mRNA and IL-1beta mRNA content did not change in the presence of 5-HT. 5-HT did not have any effect on the production of other cytokines studied. The inhibitory effect of 5-HT on TNF-alpha production was antagonized by ketanserin, a selective 5-HT(2A) antagonist, and mimicked by DOI, a selective 5-HT(2A/2C) agonist. These findings suggest that the inhibition of TNF-alpha production by 5-HT involves the participation of the 5-HT(2A) receptor subtypes in PBMC. Accordingly, we detected the presence of 5-HT(2A) receptors in PBMC by Western blot analysis. Our data support a role of 5-HT in inflammation through its effect on cytokine production in PBMC. PMID- 12578854 TI - Heat shock protein 70 associations with myelin basic protein and proteolipid protein in multiple sclerosis brains. AB - Heat shock proteins (hsp) are known to facilitate the generation of specific immune responses by chaperoning proteins and peptides involved in T cell activation. Hsp have been shown to be strikingly elevated in multiple sclerosis (MS) lesions. The unique chaperonin properties of hsp70 have allowed identification of immunogenic proteins bound to it by the ex vivo demonstration of hsp associations with proteins implicated in the immune response. We have investigated the association of hsp70 with myelin basic protein (MBP), myelin proteolipid protein (PLP) and myelin oligodendrocyte protein (MOG) in MS and control brain tissue. In co-immunoprecipitation experiments, in all samples of MS brains examined (n = 3), but not control brain tissue (n = 3), direct association of MBP with hsp70, but not with hsp90, was found. In some MS brain samples, association between PLP and hsp70 was also seen. In similar co immunoprecipitation experiments on brain tissue obtained from mice with experimental autoimmune encephalomyelitis (n = 5) induced by immunization with PLP peptide, specific association of hsp70 with PLP and MBP was found. Using surface plasmon resonance we demonstrated specific binding of hsp70 with MBP in vitro. Analysis of the amounts of MBP bound to hsp70 yielded a molecular ratio of MBP binding to hsp70 at 6.5:1. MBP complexed with hsp70 was taken up at significantly higher rates by antigen-presenting cells than MBP alone and enhanced MBP-specific immune responses. These results indicate that hsp70 specifically associates with MBP in MS brain tissue. This association might be relevant to the enhanced immune recognition of MBP in MS. PMID- 12578855 TI - IL-1-induced tumor necrosis factor-alpha elicits inflammatory cell infiltration in the skin by inducing IFN-gamma-inducible protein 10 in the elicitation phase of the contact hypersensitivity response. AB - Contact hypersensitivity (CHS) is a typical inflammatory response against contact allergens. Inflammatory cytokines, including IL-1 and tumor necrosis factor (TNF) alpha, are implicated in the reaction, although the precise roles of each cytokine have not been completely elucidated. In this report, we dissected the functional roles of IL-1 and TNF-alpha during CHS. CHS induced by 2,4,6 trinitorochlorobenzene as well as oxazolone was suppressed in both IL 1alpha/beta(-/-) and TNF-alpha(-/-) mice. Hapten-specific T cell activation, as examined by T cell proliferation, OX40 expression and IL-17 production, was reduced in IL-1alpha/beta(-/-) mice, but not in TNF-alpha(-/-) mice, suggesting that IL-1 but not TNF-alpha is required for hapten-specific T cell priming in the sensitization phase. On the other hand, TNF-alpha, induced by IL-1, was necessary for the induction of local inflammation during the elicitation phase. We also found that the expression of IFN-gamma-inducible protein 10 (IP-10) was augmented at the inflammatory site. Although IP-10 mRNA expression was abrogated in TNF alpha(-/-) mice, both CHS development and TNF-alpha mRNA expression occurred normally in IFN-gamma(-/-) mice, indicating that the induction of IP-10 during CHS was primarily controlled by TNF-alpha. Interestingly, CHS was suppressed by treatment with anti-IP-10 mAb, suggesting a critical role for IP-10 in CHS. Reduced CHS in TNF-alpha(-/-) mice was reversed by IP-10 injection during the elicitation phase. Thus, it was shown that the roles for IL-1 and TNF-alpha are different, although both cytokines are crucial for the development of CHS. PMID- 12578856 TI - Encephalitogenic activity of truncated myelin oligodendrocyte glycoprotein (MOG) peptides and their recognition by CD8+ MOG-specific T cells on oligomeric MHC class I molecules. AB - We have previously demonstrated that the 21-residue peptide pMOG(35-55) from myelin oligodendrocyte glycoprotein (MOG) contains an antigenic epitope that activates CD8(+) encephalitogenic T cells in C57BL/6 (B6) mice. To identify the core encephalitogenic epitope of CD8(+) MOG-specific T cells, we have prepared a panel of highly purified peptides of varying lengths, which span the entire length of pMOG(35-55), and tested their binding to recombinant H-2D(b) dimers and their ability to induce EAE. Two of the truncated peptides, pMOG(40-54) and pMOG(44-54), strongly bound recombinant H-2D(b) protein and this complex bound MOG-specific CD8(+) T cells. Interestingly, pMOG(40-54) retained the full capability of inducing paralytic disease, whereas only a part of the B6 mice immunized with pMOG(44-54) developed clinical paralysis and central nervous system (CNS) inflammation. Further deletion of 1 amino acid from either the N- or C-terminus of the peptide pMOG(44-54) dramatically reduced binding to recombinant H-2D(b), and abolished the induction of paralysis and CNS inflammation. Our results demonstrate that the ability of truncated pMOG(35-55) peptides to bind recombinant H-2D(b) protein does not always correlate with their ability of inducing encephalomyelitis. This approach enables the further identification of the core pathogenic epitope within the pMOG(35-55) that activates MOG-specific encephalitogenic CD8(+) T cells. PMID- 12578857 TI - Tolerance established in autoimmune disease by mating or bone marrow transplantation that target autoantigen to thymus. AB - Autoimmune diseases are a significant cause of death and morbidity, affecting up to 5% of the population. At present, there is no cure. Autologous bone marrow transplantation has been promoted as a treatment for achieving disease reversal and long-term remission. However, clinical trials in progress in Europe and North America report a significant risk of relapse. Here, we have addressed whether we can establish tolerance in an active autoimmune disease model by thymic expression of autoantigen. We show that tolerance and disease resistance can indeed be established in transgenic mice that spontaneously develop granulocyte macrophage colony stimulating factor-induced autoimmune gastritis, by mating them with disease-resistant transgenic mice that target autoantigen to the thymus. T cells from these double-transgenic mice are non-responsive to gastric antigen in vitro and fail to initiate disease following transfer to naive recipients. Further, we show that transplantation with bone marrow from disease-resistant transgenic mice renders recipient mice with gastritis tolerant to autoantigen as shown by a dramatic fall in autoantibody levels and T cell non-responsiveness to antigen in vitro. We suggest that genetically modified bone marrow targeting autoantigen to the thymus may be used to establish tolerance and prevent relapse of autoimmune disease following autologous bone marrow transplantation. PMID- 12578858 TI - Th2 bias of CD4+ NKT cells derived from multiple sclerosis in remission. AB - Although CD1d-restricted NKT cells have been implicated as a participant in the regulatory mechanism of autoimmune diseases, it remains unclear how they would regulate human autoimmune diseases such as multiple sclerosis (MS). Furthermore, although the NKT cells comprise CD4(+) and CD4(-) populations, prior studies have often represented them as simply a CD4(-) population. Given that CD4(+) and CD4( ) NKT cells may represent functionally distinct populations, it appears crucial to examine the individual NKT subset in autoimmune diseases. Here we studied the frequency and cytokine phenotypes of the CD4(+) and CD4(-) NKT cells in fresh peripheral blood mononuclear cells, and of alpha-galactosylceramide-stimulated short-term cell lines obtained during the remission or relapse phase of MS as compared with from healthy subjects (HS). Here we report that CD4(+) NKT line cells expanded from MS in remission (MS-rem) would produce a larger amount of IL 4 than those from HS or from MS in relapse (MS-rel). They were significantly biased for T(h)2 as judged by the IL-4/IFN-gamma balance. However, there was no functional bias toward T(h)1 or T(h)2 in CD4(-) NKT line cells from MS-rem due to the defects in both IFN-gamma and IL-4 production, compared with HS. Of note, although double-negative NKT cells in the periphery were greatly reduced, the reduction of CD4(+) NKT cells was only marginal, if any, in MS-rem compared with HS. The T(h)2 bias of CD4(+) NKT line cells from MS-rem may support an immunoregulatory role for the CD4(+) NKT cells in vivo. PMID- 12578859 TI - Cell type-dependent retention and transmission of HIV-1 by DC-SIGN. AB - DC-SIGN (CD209) is a C-type lectin expressed by several groups of dendritic cells (DC), including those derived from blood monocytes and DC found beneath genital epithelium. DC-SIGN binds the envelope glycoprotein of HIV-1 and facilitates transmission of infectious virus to permissive CD4(+) T cells. We have compared the capacity of DC-SIGN in different cell types to bind, retain and transmit infectious HIV-1 to T cells. The analyzed cells included monocyte-derived DC, and three different DC-SIGN-expressing transfectants termed THP, 293 and HOS. Our results show that DC-SIGN transfectants were able to bind HIV-1 virions comparably to DC. However, only the THP monocytic cell line shared with DC the capacity to retain for several days virus that was infectious for T cells. In both THP-DC-SIGN transfectants and DC, but not in 293 cells, HIV-1 was localized to intracellular compartments that did not double label for endosomal and lysosomal markers or for DC-SIGN itself. Virus remained detectable in these compartments for at least 2 days. Anti-DC-SIGN antibodies blocked the binding and transmission of HIV-1 in DC-SIGN transfectants, as monitored by PCR for HIV LTR/gag and p24 ELISA. However anti-DC-SIGN antibodies did not block virus binding and transmission to T cells as well in DC as in THP-DC-SIGN transfectants. Thus, the function of DC-SIGN in HIV-1 transmission depends on its cellular context, since only DC and the THP monocyte cell line, but not 293 and HOS, are able to use DC-SIGN to retain HIV-1 in a highly infectious state for several days. PMID- 12578860 TI - Factor VII-activating protease: coagulation, fibrinolysis, and atherothrombosis? PMID- 12578861 TI - Heart failure with a normal ejection fraction: is it really a disorder of diastolic function? PMID- 12578862 TI - Evaluation and management of diastolic heart failure. PMID- 12578863 TI - Detection of coronary artery stenoses with thin-slice multi-detector row spiral computed tomography and multiplanar reconstruction. AB - BACKGROUND: We analyzed the accuracy of multi-detector row spiral computed tomography (MDCT) using a 16-slice CT scanner with improved spatial and temporal resolution, as well as routine premedication with beta-blockers for detection of coronary stenoses. METHODS AND RESULTS: Seventy-seven patients with suspected coronary disease were studied by MDCT (12x0.75-mm cross-sections, 420 ms rotation, 100 mL contrast agent IV at 5 mL/s). Patients with a heart rate above 60/min received 50 mg atenolol before the scan. In axial MDCT images and multiplanar reconstructions, all coronary arteries and side branches with a diameter of 1.5 mm or more were assessed for the presence of stenoses exceeding 50% diameter reduction. In comparison to invasive coronary angiography, MDCT correctly classified 35 of 41 patients (85%) as having at least 1 coronary stenosis and correctly detected 57 of 78 coronary lesions (73%). After excluding 38 of 308 coronary arteries (left main, left anterior descending, left circumflex, and right coronary artery in 77 patients) classified as unevaluable by MDCT (12%), 57 of 62 lesions were detected, and absence of stenosis was correctly identified in 194 of 208 arteries (sensitivity: 92%; specificity: 93%; accuracy: 93%; positive and negative predictive values: 79% and 97%). CONCLUSIONS: MDCT coronary angiography with improved spatial resolution and premedication with oral beta-blockade permits detection of coronary artery stenoses with high accuracy and a low rate of unevaluable arteries. PMID- 12578864 TI - Marburg I polymorphism of factor VII--activating protease: a prominent risk predictor of carotid stenosis. AB - BACKGROUND: Atherothrombosis is a main pathomechanism in the evolution of vessel stenosis and is counteracted by endogenous fibrinolysis. Recently, the plasmatic serine protease "factor seven-activating protease" (FSAP) was recognized as a potent activator of prourokinase in vitro. The Marburg I polymorphism of FSAP impairs this potential and may thus facilitate arterial thrombosis. METHODS AND RESULTS: This analysis of the Bruneck Study involved 810 men and women aged 40 to 79 years. The ultrasound-based atherosclerosis progression model (5-year follow up) permits differentiation between early atherogenesis and the advanced stenotic stages of carotid artery disease. The FSAP Marburg I polymorphism was found in 37 subjects (carriage rate 4.4%). Individuals with this genetic variant showed a prominently reduced in vitro capacity to activate prourokinase. No relation was found to exist between the Marburg I polymorphism and early atherogenesis. In contrast, it emerged as a strong and independent risk predictor of incident/progressive carotid stenosis (multivariate odds ratio [95%CI], 6.6 [1.6 to 27.7]). This finding equally applied to subjects with and without co segregation of the Marburg II polymorphism. The risk profile of advanced atherogenesis further includes cigarette smoking, high lipoprotein(a), the factor V Leiden mutation, low antithrombin III, high fibrinogen, and diabetes. CONCLUSIONS: In concert with other genetic and acquired conditions known to interfere with coagulation or fibrinolysis, the Marburg I polymorphism of FSAP, which attenuates its capacity to activate prourokinase, is a significant risk predictor for the evolution and progression of carotid stenosis. PMID- 12578865 TI - Reciprocal association of C-reactive protein with adiponectin in blood stream and adipose tissue. AB - BACKGROUND: High-sensitive C-reactive protein (hs-CRP) is a well-known risk factor for coronary artery disease (CAD). Recently, we have demonstrated that adiponectin served as an antiatherogenic plasma protein which was secreted specifically from adipocytes. The present study investigated the association between adiponectin and CRP in the blood stream and adipose tissue. METHODS AND RESULTS: We studied a total of 101 male patients, 71 of whom had angiographically documented coronary atherosclerosis. As a control group, 30 patients with normal coronary angiogram were included. The plasma hs-CRP levels were negatively correlated with the plasma adiponectin levels (r=-0.29, P<0.01). The plasma adiponectin concentrations were significantly lower and the hs-CRP levels were significantly higher in the CAD patients compared with control subjects. The mRNA levels of CRP and adiponectin were analyzed by quantitative real-time polymerase chain reaction method. We found that the CRP mRNA was expressed in human adipose tissue. A significant inverse correlation was observed between the CRP and adiponectin mRNA levels in human adipose tissue (r=-0.89, P<0.01). In addition, the CRP mRNA level of white adipose tissue in adiponectin deficient mice was higher than that of wild-type mice. CONCLUSIONS: The reciprocal association of adiponectin and CRP levels in both human plasma and adipose tissue might participate in the development of atherosclerosis. PMID- 12578866 TI - Aging and the exercise pressor reflex in humans. AB - BACKGROUND: Blood flow limitation to exercising muscles engages the muscle reflex during exercise, evoking an increase in heart rate (HR), blood pressure (BP), and muscle sympathetic nerve activity (MSNA). METHODS AND RESULTS: In the current study, we examined forearm flow and autonomic responses to ischemic handgrip in young and older subjects. We studied 6 younger subjects (mean age 23.5+/-2.2 years) and 7 older subjects (mean age 65.0+/-2.4 years). Subjects performed rhythmic handgrip (thirty 1-sec contractions/min) at 30% maximal voluntary contraction during six 1-minute stages: freely perfused exercise (E1) and exercise with forearm pressure of +10, +20, +30, +40, and +50 mm Hg (E2 through E6). We measured HR, BP, MSNA, forearm flow velocity, forearm venous oxygen saturation, H(+), and lactate. Compared with E1, ischemic exercise (E2 through E6) increased HR, BP, and MSNA, reduced forearm velocity, lowered venous oxygen saturation, and raised venous lactate and H(+). Compared with the younger subjects, the older subjects had attenuated BP at E6, attenuated MSNA indices (%(Delta)bursts, bursts/100 heart beats and signal averaged MSNA), attenuated H(+) at E6, a trend toward higher levels of oxygen saturation, and similar forearm velocity and HR responses. CONCLUSIONS: Aging attenuates the muscle reflex. PMID- 12578867 TI - Osteopontin transgenic mice fed a high-cholesterol diet develop early fatty streak lesions. AB - BACKGROUND: Osteopontin (OPN) is a noncollagenous adhesion protein found at the site of atherosclerotic lesions. However, it has not yet been clarified whether or not OPN can promote atherosclerotic lesions. METHODS AND RESULTS We investigated the contribution of OPN to atherosclerosis by evaluating aortic sinus lesions of both OPN transgenic (Tg) and non-Tg mice fed an atherogenic diet (1.25% cholesterol) for 16 weeks. The atherosclerotic lesions were found to be significantly larger in OPN-Tg compared with those in non-Tg (17,859+/-2010 versus 6469+/-485 micro m(2), P<0.01). The lesions in both mice were fatty-streak lesions with an accumulation of mononuclear cells and lipids. We next investigated the production of interleukin (IL)-10 by macrophages from both mice. Compared with the non-Tg mice, a 42% (P<0.01) and 73% (P<0.001) decrease in the IL-10 production was identified in the OPN-Tg mice either without or with lipopolysaccharide. CONCLUSIONS: The expression of OPN induces fatty-streak lesion formation in mice fed an atherogenic diet and inhibits IL-10 production by macrophages, thus suggesting that OPN plays an important role in the development of fatty-streak lesions in vivo. PMID- 12578868 TI - Cardioprotective effect of diazoxide is mediated by activation of sarcolemmal but not mitochondrial ATP-sensitive potassium channels in mice. AB - BACKGROUND: We recently demonstrated that the sarcolemmal ATP-sensitive potassium (sarcK(ATP)) channel plays a key role in cardioprotection against ischemia/reperfusion injuries in Kir6.2-knockout (KO) mice. In the present study, we evaluated the effects of diazoxide, a mitochondrial ATP-sensitive potassium (mitoK(ATP)) channel opener, on ischemia-induced myocardial stunning in sarcK(ATP) channel-deficient mice. METHODS AND RESULTS: Langendorff-perfused hearts of wild-type (WT) and KO mice were subjected to global ischemia/reperfusion. Diazoxide improved the recovery of contractile function in WT hearts but not in KO hearts. Treatment with HMR1098 (a sarcK(ATP) channel blocker) but not 5-hydroxydecanoate (a mitoK(ATP) channel blocker) abolished the cardioprotective effect of diazoxide in WT hearts. In coronary-perfused WT ventricular muscle preparations, action potential shortening during ischemia was accelerated in the presence of diazoxide. CONCLUSIONS: Diazoxide enhances action potential shortening during ischemia by activating sarcK(ATP) channels and provides cardioprotection in mouse hearts. PMID- 12578869 TI - Cardiorenal and humoral properties of a novel direct soluble guanylate cyclase stimulator BAY 41-2272 in experimental congestive heart failure. AB - BACKGROUND: BAY 41-2272 is a recently introduced novel orally available agent that directly stimulates soluble guanylate cyclase (sGC) and sensitizes it to its physiological stimulator, nitric oxide. To date, its therapeutic actions in congestive heart failure (CHF) remain undefined. We characterized the cardiorenal actions of intravenous BAY 41-2272 in a canine model of CHF and compared it to nitroglycerin (NTG). METHODS AND RESULTS: CHF was induced by rapid ventricular pacing for 10 days. Cardiorenal and humoral function were assessed at baseline and with administration of 2 doses of BAY 41-2272 (2 and 10 micro g x kg(-1) x min(-1); n=8) or NTG (1 and 5 micro g x kg(-1) x min(-1); n=6). Administration of 10 micro g x kg(-1) x min(-1) BAY 41-2272 reduced mean arterial pressure (113+/-8 to 94+/-6 mm Hg; P<0.05), pulmonary artery pressure (29+/-2 to 25+/-2 mm Hg; P<0.05), and pulmonary capillary wedge pressure (25+/-2 to 20+/-2 mm Hg; P<0.05). Cardiac output (2.1+/-0.2 to 2.3+/-0.2 L/min; P<0.05) and renal blood flow (131+/ 17 to 162+/-18 mL/min; P<0.05) increased. Glomerular filtration rate was maintained. There were no changes in plasma renin activity, angiotensin II, or aldosterone. NTG mediated similar hemodynamic changes and additionally decreased right atrial pressure and pulmonary vascular resistance. CONCLUSION: The new sGC stimulator BAY 41-2272 potently unloaded the heart, increased cardiac output, and preserved glomerular filtration rate without activation of the renin-angiotensin aldosterone system in experimental CHF. These beneficial properties make direct sGC stimulation with BAY 41-2272 a promising new strategy for the treatment of cardiovascular diseases such as CHF. PMID- 12578871 TI - Association of polymorphisms of the apolipoprotein(a) gene with lipoprotein(a) levels and myocardial infarction. AB - BACKGROUND: Serum lipoprotein(a) [Lp(a)] concentration is largely determined by variability at the apolipoprotein(a) gene locus. Most prominent effects relate to polymorphisms in the promoter (a pentanucleotide [PN] repeat) and coding regions (a kringle IV [K4] repeat), the latter of which also affects Lp(a) particle size. The impact of these polymorphisms on cardiovascular risk is poorly understood. METHODS AND RESULTS: We studied both polymorphisms and Lp(a) levels in 834 registry-based myocardial infarction (MI) patients (38% women) and 1548 population-based controls. Lp(a) concentrations were inversely related with the numbers of K4 and PN repeats. However, the effect of the PN polymorphism was restricted to subjects producing small Lp(a) particles (8 PN 8.7 mg/dL; P<0.0001). The odds to present with MI were elevated in individuals producing small Lp(a) particles (75th percentile (corresponding to the 90th percentile in the healthy volunteers) was associated with an increased risk of death or myocardial infarction through 10 months of follow-up (adjusted hazard ratio, 1.53; 95% CI, 1.09 to 2.14; P=0.01). CONCLUSIONS: In a large cohort of patients with acute coronary syndromes, an elevated baseline level of MCP-1 was associated both with traditional risk factors for atherosclerosis as well as an increased risk for death or myocardial infarction, independent of baseline variables. Because it appears to play a crucial role at multiple stages of atherosclerosis, MCP-1 is attractive as a surrogate biomarker and merits further study as a potential therapeutic target. PMID- 12578872 TI - Detecting occult coronary disease in a high-risk asymptomatic population. AB - BACKGROUND: Exercise stress testing alone or with perfusion imaging is the standard screening method to determine the presence of obstructive coronary artery disease (CAD) in people with chest pain. In asymptomatic individuals with a family history of premature CAD, it is unclear whether abnormalities on these functional exercise tests represent significant coronary disease. METHODS AND RESULTS: An abnormal exercise test, thallium scan, or both occurred in 153 (21%) of 734 asymptomatic siblings of persons with documented CAD, of whom 105 underwent coronary angiography with quantitative analysis of stenosis severity. Overall, 95% had coronary atherosclerosis, but only 39% had 1 or more stenoses with >or=50% narrowing. Of 30 siblings in whom the exercise test and perfusion scan were both abnormal, 70% had >or=50% stenoses. The mean stenosis in arteries that fed perfusion defects was only 43+/-31%, and 68% of such stenoses were <50%. However, in 71% of all defects, the location matched arteries with the most severe stenoses. CONCLUSIONS: In asymptomatic persons with a family history of CAD, abnormal exercise scintigraphy identifies predominantly mild coronary atherosclerosis. Perfusion defects may be caused by coronary vasomotor dysfunction in addition to atherosclerotic plaque. PMID- 12578873 TI - Low-T3 syndrome: a strong prognostic predictor of death in patients with heart disease. AB - BACKGROUND: Clinical and experimental data have suggested a potential negative impact of low-T3 state on the prognosis of cardiac diseases. The aim of the present prospective study was to assess the role of thyroid hormones in the prognosis of patient population with heart disease. METHODS AND RESULTS: A total of 573 consecutive cardiac patients underwent thyroid function profile evaluation. They were divided in two subgroups: group I, 173 patients with low T3, ie, with free T3 (fT3) <3.1 pmol/L, and group II, 400 patients with normal fT3 (>or=3.1 pmol/L). We considered cumulative and cardiac death events. During the 1-year follow-up, there were 25 cumulative deaths in group I and 12 in group II (14.4% versus 3%, P<0.0001); cardiac deaths were 13 in group I and 6 in group II (7.5% versus 1.5%, P=0.0006). According to the Cox model, fT3 was the most important predictor of cumulative death (hazard ratio [HR] 3.582, P<0.0001), followed by dyslipidemia (HR 2.955, P=0.023), age (HR 1.051, P<0.005), and left ventricular ejection fraction (HR 1.037, P=0.006). At the logistic multivariate analysis, fT3 was the highest independent predictor of death (HR 0.395, P=0.003). A prevalence of low fT3 levels was found in patients with NYHA class III-IV illness compared with patients with NYHA class I-II (chi(2) 5.65, P=0.019). CONCLUSIONS: Low-T3 syndrome is a strong predictor of death in cardiac patients and might be directly implicated in the poor prognosis of cardiac patients. PMID- 12578874 TI - Combined ventricular systolic and arterial stiffening in patients with heart failure and preserved ejection fraction: implications for systolic and diastolic reserve limitations. AB - BACKGROUND: Heart failure with preserved ejection fraction (HF-nlEF) is common in aged individuals with systolic hypertension and is frequently ascribed to diastolic dysfunction. We hypothesized that such patients also display combined ventricular-systolic and arterial stiffening that can exacerbate blood pressure lability and diastolic dysfunction under stress. METHODS AND RESULTS: Left ventricular pressure-volume relations were measured in patients with HF-nlEF (n=10) and contrasted with asymptomatic age-matched (n=9) and young (n=14) normotensives and age- and blood pressure-matched controls (n=25). End-systolic elastance (stiffness) was higher in patients with HF-nlEF (4.7+/-1.5 mm Hg/mL) than in controls (2.1+/-0.9 mm Hg/mL for normotensives and 3.3+/-1.0 mm Hg/mL for hypertensives; P<0.001). Effective arterial elastance was also higher (2.6+/-0.5 versus 1.9+/-0.5 mm Hg/mL) due to reduced total arterial compliance; the latter inversely correlated with end-systolic elastance (P=0.0001). Body size and stroke volumes were similar and could not explain differences in ventricular-arterial stiffening. HF-nlEF patients also displayed diastolic abnormalities, including higher left ventricular end-diastolic pressures (24.3+/-4.6 versus 12.9+/-5.5 mm Hg), caused by an upward-shifted diastolic pressure-volume curve. However, isovolumic relaxation and the early-to-late filling ratio were similar in age- and blood pressure-matched controls. Ventricular-arterial stiffening amplified stress-induced hypertension, which worsened diastolic function, and predicted higher cardiac energy costs to provide reserve output. CONCLUSION: Patients with HF-lnEF have systolic-ventricular and arterial stiffening beyond that associated with aging and/or hypertension. This may play an important pathophysiological role by exacerbating systemic load interaction with diastolic function, augmenting blood pressure lability, and elevating cardiac metabolic demand under stress. PMID- 12578875 TI - Identification of serum soluble ST2 receptor as a novel heart failure biomarker. AB - BACKGROUND: Using genomic technology, we previously identified an interleukin-1 receptor family member, ST2, as a gene markedly induced by mechanical strain in cardiac myocytes. The soluble receptor form of ST2 is secreted and detectable in human serum. This study tested the hypothesis that soluble ST2 levels in the serum of patients with severe chronic heart failure are increased in patients with neurohormonal activation. METHODS AND RESULTS: Serum samples, clinical variables, and neurohormone levels from the PRAISE-2 heart failure trial (NYHA functional class III-IV; end point, mortality or transplantation) were analyzed. ST2 serum measurements were performed with ELISA on samples from 161 patients obtained at trial enrollment and from 139 of the same patients obtained 2 weeks after trial enrollment. Baseline ST2 levels were correlated with baseline B-type natriuretic peptide (BNP) levels (r=0.36, P<0.0001), baseline proatrial natriuretic peptide (ProANP) levels (r=0.36, P<0.0001), and baseline norepinephrine levels (r=0.39, P<0.0001). The change in ST2 was significant as a univariate predictor of subsequent mortality or transplantation (P=0.048), as was baseline BNP (P<0.0001) and baseline ProANP (P<0.0001). In multivariate models including BNP and ProANP, the change in ST2 remained significant as a predictor of mortality or transplantation independent of BNP and ProANP. CONCLUSIONS: Serum soluble ST2 is a novel biomarker for neurohormonal activation in patients with heart failure. In patients with severe chronic NYHA class III to IV heart failure, the change in ST2 levels is an independent predictor of subsequent mortality or transplantation. PMID- 12578876 TI - Central sleep apnea in left ventricular dysfunction: prevalence and implications for arrhythmic risk. AB - BACKGROUND: The prevalence and characteristics of sleep-disordered breathing in patients with asymptomatic left ventricular (LV) dysfunction are unknown. Therefore, we evaluated the prevalence of sleep-disordered breathing in patients with LV dysfunction without overt heart failure and tested the hypothesis that sleep-disordered breathing is linked to greater hemodynamic and autonomic impairment. METHODS AND RESULTS: We studied 47 patients with LV ejection fractions or=15/h, was present in 26 patients (55%), 17 (36%) of whom had severe CSA (apnea-hypopnea index >or=30/h). Obstructive sleep apnea was evident in 5 patients (11%). The prevalence and severity of CSA were higher in patients with ischemic cardiomyopathy than in patients with nonischemic cardiomyopathy (P<0.05). Exercise tolerance and echocardiographic indices of systolic and diastolic function were similar in patients without CSA, with mild CSA, and with severe CSA. Heart rate variability was markedly depressed in patients with CSA (P<0.05). Patients with severe CSA also had a higher incidence of nonsustained ventricular tachycardia (P=0.05). CONCLUSIONS: CSA is highly prevalent in patients with asymptomatic LV dysfunction. The severity of CSA may not be related to the severity of hemodynamic impairment. Severe CSA is associated with impaired cardiac autonomic control and with increased cardiac arrhythmias. PMID- 12578877 TI - Characterization of left atrial activation in the intact human heart. AB - BACKGROUND: The patterns of activation of the human left atrium (LA), how they relate to atrial myocardial architecture, and their role in arrhythmogenesis remain largely unknown. METHODS AND RESULTS: Left atrial endocardial activation was mapped in 19 patients with a percutaneous noncontact mapping system. Earliest endocardial breakthrough during sinus rhythm (SR) occurred more frequently in the septal (63%, principally posteroseptal) than anterosuperior (37%) LA and varied little with isoproterenol or high right atrial pacing rate. Regardless of site of breakthrough, LA activation was characterized in all patients by propagation around a variably complete line of functional conduction block, descending on the posterior wall from the roof, passing between the ostia of the superior and then inferior pulmonary veins (PVs) before turning septally, passing below the oval fossa, and merging further anteriorly with the septal mitral annulus. Examination of the myocardial architecture in 10 normal adult postmortem hearts revealed an abrupt change in subendocardial fiber orientation along a line following the same course. During episodes of focal initiation of atrial fibrillation (AF), interaction was observed between wavefronts entering the LA from PVs and this functional line of conduction block that resulted in LA macroreentry or formation of daughter wavefronts. CONCLUSIONS: The LA endocardium has complex but characteristic patterns of activation during sinus rhythm, pacing, and AF initiation by PV ectopy that are determined largely by the functional properties of atrial musculature. These findings have important implications for both pacing and ablative strategies for the prevention of initiation of AF. PMID- 12578879 TI - Blood pressure, risk of ischemic cerebrovascular and ischemic heart disease, and longevity in alpha(1)-antitrypsin deficiency: the Copenhagen City Heart Study. AB - BACKGROUND: Because elastase in alpha(1)-antitrypsin deficiency may attack elastin in the arterial wall, we tested whether alpha(1)-antitrypsin deficiency is associated with reduced blood pressure, risk of ischemic cerebrovascular (ICVD) and ischemic heart disease (IHD), and longevity. METHODS AND RESULTS: We genotyped 7963 control subjects from the adult general population of Denmark, 1131 Danish patients with ICVD, and 2221 Danish patients with IHD. Compared with MM/MS individuals, systolic blood pressure was lower by 15 mm Hg in ZZ homozygotes (n=6, P=0.03) and 9 mm Hg in MZ heterozygotes with IHD (n=39, P=0.02). Odds ratios for ICVD and IHD in MZ versus MM/MS individuals were 0.70 (0.51 to 0.96) and 0.77 (0.61 to 0.98). Finally, mean ages of MZ and MM/MS control subjects were 58 and 56 years (Mann-Whitney: P=0.008), and relative alpha(1)-antitrypsin MZ genotype frequencies increased from 20 to 93 years among control subjects (chi(2), P=0.002). CONCLUSIONS: ZZ alpha(1)-antitrypsin deficiency and MZ intermediate deficiency in the context of IHD are associated with reduced blood pressure, and MZ is associated with reduced risk of ICVD and IHD. Because MZ heterozygosity was associated with increased age, MZ heterozygosity could be a beneficial condition. PMID- 12578878 TI - Effect of epicardial or biventricular pacing to prolong QT interval and increase transmural dispersion of repolarization: does resynchronization therapy pose a risk for patients predisposed to long QT or torsade de pointes? AB - BACKGROUND: The present study examined pacing site-dependent changes in QT interval and transmural dispersion of repolarization (TDR) and their potential role in the development of torsade de pointes (TdP). METHODS AND RESULTS: In humans, the QT interval, JT interval, and TDR were measured in 29 patients with heart failure during right ventricular endocardial pacing (RVEndoP), biventricular pacing (BiVP), and left ventricular epicardial pacing (LVEpiP). In animal experiments, pacing site--dependent changes in ventricular repolarization were examined with a rabbit left ventricular wedge preparation in which action potentials from endocardium and epicardium could be simultaneously recorded with a transmural ECG. In humans, LVEpiP and BiVP led to significant QT and JT prolongation. LVEpiP also enhanced TDR. Frequent R-on-T extrasystoles generated by BiVP and LVEpiP but completely inhibited by RVEndoP occurred in 4 patients, of whom 1 developed multiple episodes of nonsustained polymorphic ventricular tachycardia and another suffered incessant TdP. In rabbit experiments, switching from endocardial to epicardial pacing produced a net increase in QT interval and TDR by 17+/-5 and 22+/-5 ms, respectively (n=6, P<0.01), without parallel increases in ventricular transmembrane action potential durations. Epicardial pacing facilitated transmural propagation of early afterdepolarization, leading to the development of R-on-T extrasystoles and TdP in the presence of action potential duration-prolonging agents. CONCLUSIONS: LVEpiP and BiVP increase QT, JT, and TDR by altering the transmural sequence of activation of the intrinsically heterogeneous ventricular myocardium. Our data suggest that the resultant exaggeration of arrhythmic substrates can lead to the development of TdP in a subset of patients. PMID- 12578880 TI - Intensive blood pressure control reduces the risk of cardiovascular events in patients with peripheral arterial disease and type 2 diabetes. AB - BACKGROUND: Peripheral arterial disease (PAD) and diabetes are both associated with a high risk of ischemic events, but the role of intensive blood pressure control in PAD has not been established. METHODS AND RESULTS: The Appropriate Blood Pressure Control in Diabetes study followed 950 subjects with type 2 diabetes for 5 years; 480 of the subjects were normotensive (baseline diastolic blood pressure of 80 to 89 mm Hg). Patients randomized to placebo (moderate blood pressure control) had a mean blood pressure of 137+/-0.7/81+/-0.3 mm Hg over the last 4 years of treatment. In contrast, patients randomized to intensive treatment with enalapril or nisoldipine had a mean 4-year blood pressure of 128+/ 0.8/75+/-0.3 mm Hg (P<0.0001 compared with moderate control). PAD, which is defined as an ankle-brachial index <0.90 at the baseline visit, was diagnosed in 53 patients. In patients with PAD, there were 3 cardiovascular events (13.6%) on intensive treatment compared with 12 events (38.7%) on moderate treatment (P=0.046). After adjustment for multiple cardiovascular risk factors, an inverse relationship between ankle-brachial index and cardiovascular events was observed with moderate treatment (P=0.009), but not with intensive treatment (P=0.91). Thus, with intensive blood pressure control, the risk of an event was not increased, even at the lowest ankle-brachial index values, and was the same as in a patient without PAD. CONCLUSIONS: In PAD patients with diabetes, intensive blood pressure lowering to a mean of 128/75 mm Hg resulted in a marked reduction in cardiovascular events. PMID- 12578881 TI - Statin use and leg functioning in patients with and without lower-extremity peripheral arterial disease. AB - BACKGROUND: We determined whether statin use (versus nonuse) is associated with superior lower-extremity functioning independently of cholesterol levels and other confounders in patients with and without peripheral arterial disease. METHODS AND RESULTS: Participants included 392 men and women with an ankle brachial index (ABI) <0.90 and 249 with ABI 0.90 to 1.50. Functional outcomes included 6-minute walk distance and 4-meter walking velocity. A summary performance score combined performance in walking speed, standing balance, and time for 5 repeated chair rises into an ordinal score ranging from 0 to 12 (12=best). Adjusting for age, sex, ABI, comorbidities, education level, medical insurance status, cholesterol, and other confounders, participants taking statins had better 6-minute walk performance (1276 versus 1218 feet, P=0.045), faster walking velocity (0.93 versus 0.89 m/s, P=0.006), and a higher summary performance score (10.2 versus 9.4, P<0.001) than participants not taking statins. Positive associations were attenuated slightly after additional adjustment for C-reactive protein level but remained statistically significant for walking velocity and the summary performance score. We did not find significant associations between lower-extremity functioning and aspirin, ACE inhibitors, vasodilators, or beta-blockers. CONCLUSIONS: Statin use is associated with superior leg functioning compared with no statin use, independent of cholesterol levels and other potential confounders. These data suggest that non cholesterol-lowering properties of statins may favorably influence functioning in persons with and without peripheral arterial disease. PMID- 12578882 TI - Influenza infection exerts prominent inflammatory and thrombotic effects on the atherosclerotic plaques of apolipoprotein E-deficient mice. AB - BACKGROUND: The role of infection in the development and complications of atherosclerosis has been the focus of much attention. We reported previously that influenza vaccination was associated with reduced risk of recurrent myocardial infarction. Here, we report the effect of influenza A virus on the apolipoprotein E-deficient (apoE(-/-)) mouse, an animal model of atherosclerosis. METHODS AND RESULTS: Twenty-four apoE(-/-) mice >24 months old were injected with 1 LD(50) (lethal dose 50) of influenza A virus. Ten wild-type C57BL/6 infected mice and 11 noninfected age-matched apoE(-/-) mice served as controls. Multiple aortic sections were studied histologically 3, 5, and 10 days later. The infected mice showed markedly increased intimal cellularity compared with the noninfected apoE( /-) mice. No aortic abnormalities were seen in infected wild-type mice. Ten infected apoE(-/-) mice had a significant subendothelial infiltrate composed of a heterogeneous group of cells that stained positively for smooth muscle cell actin, F4/80 (macrophages), and CD3 (T lymphocytes). One case of subocclusive platelet and fibrin-rich thrombus was seen. CONCLUSIONS: This study shows that influenza infection promotes inflammation, smooth muscle cell proliferation, and fibrin deposition in atherosclerotic plaques. PMID- 12578884 TI - Local drug delivery via a coronary stent with programmable release pharmacokinetics. AB - BACKGROUND: Fixed drug release kinetics and vessel wall partitioning may limit the effectiveness of drug-eluting stents. We report preliminary experience using a new coronary stent with programmable pharmacokinetics. METHODS AND RESULTS: A newly designed metallic stent contains honeycombed strut elements with inlaid stacked layers of drug and polymer. In vitro studies evaluated recipes for loading paclitaxel to establish the parameters for controlling drug release. Manipulation of the layers of biodegradable polymer and drug allowed varying of the initial 24-hour burst release of paclitaxel from 69% to 8.6% (P<0.0001). Late release of drug could be adjusted dependently or independently of early burst release. A biphasic release profile was created by the addition of blank layers of polymer within the stack. In the 30-day porcine coronary model (n=17 pigs), there was a 70% reduction in late loss (0.3+/-0.5 versus 1.0+/-0.5 mm, P=0.04), a 28% increase in luminal volume (132+/-12 versus 103+/-21 mm(3), P=0.02), and a 50% decrease in histological neointimal area (2.0+/-0.5 versus 4.0+/-1.6 mm(2); P<0.001) compared with bare metal controls. Temporal and regional variations in vascular healing were seen histologically. CONCLUSIONS: Layered polymer/drug inlay stent technology permits flexible and controllable pharmacokinetic profiles. Programmable, complex chemotherapy using this approach may be feasible for the treatment of cardiovascular disease. PMID- 12578883 TI - Endothelium-derived hyperpolarizing factor in human internal mammary artery is 11,12-epoxyeicosatrienoic acid and causes relaxation by activating smooth muscle BK(Ca) channels. AB - BACKGROUND: Left internal mammary arteries (LIMAs) synthesize endothelium-derived hyperpolarizing factor (EDHF), a short-lived K(+) channel activator that persists after inhibition of nitric oxide (NO) and prostaglandin synthesis. EDHF hyperpolarizes and relaxes smooth muscle cells (SMCs). The identity of EDHF in humans is unknown. We hypothesized that EDHF (1) is 11,12-epoxyeicosatrienoic acid (11,12-EET); (2) is generated by cytochrome P450-2C, CYP450-2C; and (3) causes relaxation by opening SMC large-conductance Ca(2+)-activated K(+) channels (BK(Ca)). METHODS AND RESULTS: The identity of EDHF and its mechanism of action were assessed in 120 distal human LIMAs and 20 saphenous veins (SVs) obtained during CABG. The predominant EET synthesized by LIMAs is 11,12-EET. Relaxations to exogenous 11,12-EET and endogenous EDHF are of similar magnitudes. Inhibition of EET synthesis by chemically distinct CYP450 inhibitors (17-octadecynoic acid, N-methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide), or a selective EET antagonist (4,15-epoxyeicosa-5(Z)-enoic acid) impairs EDHF relaxation. 11,12-EET activates a BK(Ca) current and hyperpolarizes LIMA SMCs. Inhibitors of BK(Ca) but not inward-rectifier or small-conductance K(Ca) channels abolish relaxation to endogenous EDHF and exogenous 11,12-EET. BK(Ca) and CYP450-2C mRNA and proteins are more abundant in LIMAs than in SVs, perhaps explaining the lack of EDHF activity of the SV. Laser capture microdissection and quantitative RT-PCR demonstrate that BK(Ca) channels are primarily in vascular SMCs, whereas the CYP450-2C enzyme is present in both the endothelium and SMCs. CONCLUSIONS: In human LIMAs, EDHF is 11,12-EET produced by an EDHF synthase CYP450-2C and accounting for approximately 40% of net endothelial relaxation. 11,12-EET causes relaxation by activating SMC BK(Ca) channels. PMID- 12578885 TI - Beating atrial septal defect closure monitored by epicardial real-time three dimensional echocardiography without cardiopulmonary bypass. AB - BACKGROUND: We assessed the feasibility of beating atrial septal defect (ASD) closure monitored by real-time 3D echocardiography (RT3DE). METHODS AND RESULTS: RT3DE was developed with prototype ultrasound equipment consisting of a high speed 3D rendering unit with a frame rate of 5 to 10 frames/s. We also developed a prototype semiautomatic suture device and suture cutting system. In the experiment, 12 mongrel dogs were anesthetized, and after median sternotomy, the echo probe was applied directly to the surface of the right atrium. Three surgical maneuvers (balloon atrial septectomy, enlargement of the ASD, and ASD closure) were performed through the atrial port inserted into the right atrial appendage. The heart was then excised, and the area of the ASD measured by RT3DE was compared with its area measured directly. The ASD was successfully closed in all experimental animals except the first 2. Examination of the excised heart showed that none of the sutures were loose. The mean area of the ASD was 82.5+/ 38.6 mm(2) when measured by RT3DE and 81.6+/-38.2 mm(2) when measured directly, and there was a significant correlation between the areas measured by RT3DE and those measured directly (echo measurements=1.007xdirect measurements+0.337; P<0.0001). A Bland-Altman analysis revealed close agreement between the results obtained by the 2 methods (7.807 mm(2) upper and -6.024 mm(2) lower limit of agreement). CONCLUSIONS: Introduction of RT3DE, a semiautomatic suture device, and a suture cutting system made beating ASD closure without cardiopulmonary bypass possible. PMID- 12578886 TI - Image in cardiovascular medicine. Aortic xanthomatosis with coronary ostial occlusion in a child homozygous for a nonsense mutation in ABCG8. PMID- 12578887 TI - Images in cardiovascular medicine. Value of three-dimensional reconstructions in evaluating thoracic aortic aneurysms. PMID- 12578888 TI - Coronary anomalies. PMID- 12578889 TI - Transforming growth factor-beta function blocking already effective as therapeutic strategy. PMID- 12578890 TI - Effects of ventricular premature stimulus coupling interval on blood pressure and heart rate turbulence. PMID- 12578891 TI - Platelet glycoprotein IIb/IIIa inhibitors. PMID- 12578892 TI - Increased C-reactive protein and increased plasma interleukin-6 may synergistically affect the progression of coronary atherosclerosis in obstructive sleep apnea syndrome. PMID- 12578893 TI - Autonomic tone variations before the onset of paroxysmal atrial fibrillation. PMID- 12578894 TI - First selective coronary arteriogram. PMID- 12578896 TI - Quality assurance activities in radiotherapy. PMID- 12578895 TI - Keeping the pressure down in patients with type 2 diabetes and peripheral artery disease. PMID- 12578897 TI - Quality assurance of radiotherapy and its clinical assessment. AB - BACKGROUND: We investigated the clinical quality assurance (QA) of radiotherapy in Japan since 1981. The aim of this study was to establish the QA of a radiotherapy system and its clinical assessment in Japan. METHODS: We introduced the Patterns of Care Study (PCS) into Japan to perform this study in 1996. The PCS is a retrospective study designed to establish the national practice for cancer patients during a specific period and should be a complementary study to a prospective randomized controlled study. We collected precise data for 4399 patients with carcinomas of the breast, cervix, esophagus, lung and prostate by means of external audits for 96 institutes from 1998 through 2001. Patients were randomly sampled with two-stage cluster sampling. We stratified 556 institutes into four categories according to the academic condition and annual number of radiotherapy patients. National and regional averages of various factors of radiotherapy could be calculated and were used to measure QA of radiotherapy. RESULTS: Using a standard score, we could compare the process of individual institutions with national averages and feed back the evaluation score to each institution. With a PCS process survey, we could observe the dissemination of the treatment method under evidence-based medicine from the prospective randomized controlled study. We proposed future prediction of the number of radiotherapy patients and a counter plan for equipment and personnel. The first US-Japan PCS Workshop was held at San Francisco in 2001. CONCLUSION: We could establish QA of a radiotherapy system using PCS 1995-97 in Japan. PMID- 12578898 TI - Clinical factors associated with suicidality in cancer patients. AB - BACKGROUND: Previous epidemiological studies have indicated that the risk of suicide in cancer patients is higher than that of the general population. In addition, euthanasia and physician-assisted suicide (PAS) have recently become controversial medical, ethical and legal issues all over the world. Although suicide in cancer patients and appropriate management of cancer patients with suicidality are critical issues in clinical oncology practice, there have been very few studies to understand suicidality in cancer patients. The purpose of this study was to explore the clinical factors associated with suicidality in Japanese patients with cancer. METHODS: We investigated the clinical factors associated with suicidality in cancer patients by analyzing the consultation data of patients referred to the Psychiatry Division, National Cancer Centre Hospital and Hospital East, Japan. RESULTS: Of 1713 psychiatric referrals, 62 (3.6%) were related to suicidality, including 44 cases with suicidal ideation, 10 suicide attempts and eight cases who had requested euthanasia and/or continuous sedation. Most of the patients suffered from physical distress and/or psychiatric disorders. The results of a multivariate analysis comparing cancer patients with a psychiatric referral related to suicidality and those referred for other reasons indicated that impaired physical functioning and major depression were significant associated factors. CONCLUSIONS: Our findings suggest that early detection and appropriate management of major depression and comprehensive care improving physical functioning may help to prevent suicide and manage suicidality in Japanese cancer patients. PMID- 12578899 TI - A phase I study of carboplatin and docetaxel for advanced non-small cell lung cancer using the continual reassessment method. AB - PURPOSE: This phase I study was designed to determine the maximum tolerated dose of carboplatin combined with a fixed dose of docetaxel (60 mg/m(2)) and the safety and efficacy of this combination chemotherapy in unresectable non-small cell lung cancer. METHODS: Patients received a 60 min intravenous infusion of docetaxel followed by a 90 min infusion of carboplatin with dose escalation using the continual reassessment method. The starting dose of carboplatin was targeted to an area under the plasma concentration curve of 3 using Calvert's equation and dose escalation was based on course 1 toxicities. RESULTS: From January 1999 to February 2000, 16 patients entered this trial. The major dose-limiting toxicity was neutropenia. Thrombocytopenia was rare and major non-hematological toxicities included fever that was not associated with neutropenia and grade 2 nausea and vomiting. Objective responses were seen in five patients (response rate 31.3%). CONCLUSIONS: Based on this phase I clinical trial, the maximum tolerated dose of carboplatin combined with 60 mg/m(2) of docetaxel was a target area under the plasma concentration curve (tAUC) of 6 and the recommended tAUC for further trials is 5.5. This combination appeared to be effective for non-small cell lung cancer. A phase II clinical trial is recommended using 60 mg/m(2) of docetaxel and carboplatin with a tAUC of 5.5. PMID- 12578900 TI - Indications for sentinel lymph node biopsy in patients with breast cancer: retrospective and simulation analyses. AB - BACKGROUND: The disease status of axillary lymph nodes at diagnosis is the most powerful predictor of prognosis for patients with breast cancer. Axillary lymph node dissection (ALND) has been the standard treatment. Recently, lymphatic mapping and sentinel lymph node biopsy (SNB) have been suggested as alternatives to ALND. However, a clear indication for SNB has not yet been established. The goal of this study was to determine which patients might best benefit from the SNB method. METHODS: A retrospective study compared the relationship of age, menopausal status, tumor size group, histology and clinical evaluation of nodes to histological axillary lymph node status in 5892 consecutive Japanese breast cancer patients who underwent ALND. A simulation analysis with reported SNB sensitivity and specificity was carried out. RESULTS: Clinical lymph node evaluation was the most relevant factor to predict axillary nodal status. However, even patients with no clinical nodal involvement (N0) showed only 71% axillary nodal negativity. Tumor group based on tumor size and histology also related to axillary nodal status. The simulation analysis revealed that of patients with negative lymph node probability, more than 66% were good candidates for SNB. CONCLUSION: The clinically negative node (N0) is the most important indication for SNB. Therefore, the candidates for SNB should be picked from the patients with N0. When no histological information is available, patients whose tumor is 20 mm or smaller are candidates for SNB. When histological information is available, the SNB indication extends to patients whose type of tumor has histologically good prognosis or a low probability of having nodal involvement, regardless of tumor size. PMID- 12578901 TI - Expression of MUC1 and MUC2 mucin gene products in human ovarian carcinomas. AB - BACKGROUND: Aberrations in expression of mucin glycoproteins have been observed during malignant transformation of human ovarian epithelium. To date, several secretory mucin genes designated the MUC gene family have been identified, of which MUC1 encodes a mammary-type and MUC2 an intestinal-type epithelial mucin. However, information on the expression and potential value of MUC1 and MUC2 mucins in ovarian cancer is limited. METHODS: This study investigated immunohistochemical expressions of MUC1 and MUC2 mucins in 23 benign and 45 malignant human ovarian tumors to assess their clinicopathological relevance. RESULTS: All benign serous tumors and also associated normal-appearing epithelia expressed MUC1 mucin on the cell surfaces. Benign mucinous tumors occasionally expressed MUC1 and MUC2 mucins. Most serous carcinomas (19/21; 90%) expressed MUC1 but not MUC2 mucin. Of the 16 mucinous carcinomas, 10 (62%) and five (31%) expressed MUC1 and MUC2 mucins, respectively. Four of the five clear cell and the three endometroid type carcinomas expressed MUC-1 but not MUC-2 mucin. A significant association was found between a high expression of MUC1 and histological grade (P = 0.005) and also disease stage (P = 0.001). CONCLUSION: These results suggest that a high expression of MUC1 may contribute to a poor prognosis in ovarian carcinoma. PMID- 12578902 TI - Artificial neural network analysis for predicting pathological stage of clinically localized prostate cancer in the Japanese population. AB - BACKGROUND: Although prostate cancer has been prevalent in Japan, there has been no particular model for predicting the pathological stage in the Japanese population. We examined whether artificial neural network analysis (ANNA), which is a relatively new diagnostic tool in prostate cancer, can be one of the predictive methods for predicting organ confinement, compared with the traditional logistic regression model, in the Japanese population for the first time. METHODS: The study population comprised 178 men who underwent radical prostatectomy at our institutions between October 1992 and May 1999. As additional pretreatment parameters to the preoperative serum PSA level, clinical TNM classification and biopsy Gleason score, the percentage of number of cores exhibiting traces of tumor, maximum tumor length in biopsy cores, PSA density and patient age were used. The predictive ability of ANNA with several parameters for a set of 36 randomly selected test data was compared with those of logistic regression analysis and 'Partin Tables' by area under the receiver operating characteristics (ROC) curve analysis. RESULTS: Of 178 patients, 97 (54.5%) had organ-confined disease but 81 (45.5%) had locally advanced disease. With three parameters, the area under the ROC curve of ANNA (0.825 +/- 0.071) was larger than those for logistic regression (0.782 +/- 0.079) and Partin Tables (0.756 +/- 0.087), but not to a significant extent (P = 0.690 and 0.541). Although the expansion of the parameters did not increase the difference in area under the ROC curve between the best ANNA and logistic regression (0.899 +/- 0.053 and 0.873 +/ 0.065, respectively), the difference between the best ANNA and Partin Tables did not reach but approached statistical significance (P = 0.157). CONCLUSION: Although more modeling optimization is necessary to improve the predictive accuracy and generalizability of ANNA, we suggest that there is the possibility for this new predictive method to evolve in the analysis of clinical staging of prostate cancer. PMID- 12578903 TI - Predicting prostate specific antigen failure after radical retropubic prostatectomy for T1c prostate cancer. AB - PURPOSE: Clinicopathological data were reviewed to find a predictor of prostate specific antigen (PSA) failure in Taiwanese patients who had received radical retropubic prostatectomy (RRP) for stage T1c prostate cancer (PC). METHODS: Fifty five consecutive men who underwent RRP for stage T1c PC were included. The clinical end point was PSA failure (PSA >0.2 ng/ml). Preoperative PSA, free-to total PSA ratio, prostate volume, PSA density, transrectal sextant biopsy and whole mount of RRP parameters were analyzed for their ability to predict postoperative PSA failure. RESULTS: Fifteen of the 55 patients developed PSA failure during the follow-up period. Those with PSA failure had higher PSA, higher percentage of cancer in biopsies and higher biopsy Gleason score than the freedom from PSA failure group (all P < 0.05). The PSA failure group had higher pathology Gleason score and a higher incidence of extracapsular tumor extension than the freedom from PSA failure group (all P < 0.05). The PSA failure group had a larger tumor volume and higher incidence of combined peripheral lobe with transitional lobe involvement than the freedom from PSA failure group (all P < 0.05). Multivariate analysis revealed that the predictors for PSA failure after RRP were biopsy Gleason score > or =6, tumor volume > or =2.5 ml and PSA > or =10 ng/ml. CONCLUSION: The single most significant predictor for PSA failure in T1c PC patients after RRP was tumor volume > or =2.5 ml. PMID- 12578904 TI - Reactivation of hepatitis B virus infection in pancreatic cancer: a case report. AB - Impairment of liver function can be seen in cancer patients for a variety of reasons including metastasis, biliary obstruction, toxicity of chemotherapy or other concomittant drugs, can be paraneoplastic or can be due to infections, especially with hepatotropic viruses. Mostly reported in hematological malignancies, reactivation of hepatitis B virus (HBV) is a complication of chronic HBV infection in patients under cytotoxic or immunosuppressive therapy. Up to now, only a few cases of hepatitis B reactivation have been described in patients with solid tumors. We report an acute reversible deterioration of liver function tests following cytotoxic chemotherapy in an HBsAg-positive patient with metastatic pancreatic cancer due to hepatitis B reactivation. As far as we know, this is the first case of pancreatic carcinoma with reactivation following chemotherapy. As experience with acute flares of HBV carriers in solid tumors increases, the establishment of guidelines for the management of these patients has become more crucial. PMID- 12578905 TI - Enteropathy-type T-cell lymphoma showing repeated small bowel rupture and refractoriness to chemotherapy: a case report. AB - The majority of gastrointestinal lymphomas arise in the stomach, whereas lymphomas occurring in the intestine are relatively rare and a limited fraction of them show the T-cell phenotype with clinical manifestations similar to de novo celiac disease. Enteropathy-type T-cell lymphoma is extremely rare in Japan, presumably owing to the very low incidence of celiac disease among the Japanese population. Here, we report a 66-year-old Japanese male who was diagnosed as having enteropathy-type T-cell lymphoma preceded by diarrhea and intermittent bloody stool for over 1 year. He was admitted to our hospital as an emergency case because of panperitonitis due to intestinal perforation. After immediate partial small-bowel resection, cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) chemotherapy was started. However, the disease was highly refractory and was exacerbated with leukemic transformation. Subsequent salvage chemotherapy could not be completed because of the formation of spontaneous jejuno-abdominal fistula, followed by fatal septic shock. Particular attention should be paid to the peculiar clinical manifestations of enteropathy-type T-cell lymphoma such as malnutrition, frequent intestinal perforation and refractoriness to chemotherapy. PMID- 12578906 TI - Rationale and study design of dietary intervention in patients polypectomized for tumors of the colorectum. AB - We have implemented a randomized controlled dietary intervention in patients polypectomized for tumors of the colorectum to elucidate potential beneficial effects of n-3 polyunsaturated fatty acids (PUFAs) on the development of colorectal tumors. Those individuals in the experimental group were advised not only to decrease their consumption of fats/oils as a whole and foods supplying n 6 PUFAs but also to increase intake of foods and supplements containing n-3 PUFAs, while those in the comparison group were cautioned to reduce intake of fats/oils as a whole. Patients' compliance/adherence was monitored with a semi quantitative food frequency questionnaire and by assessment of fatty acid concentrations in plasma, membranes of red blood cells and sigmoid colon samples. As for endpoints to assess tumor suppressive effects of n-3 PUFAs, the number/multiplicity, sizes and incidence rates of colorectal tumors were compared between the experimental and comparison groups after 12 and 24 months of the dietary intervention. On the specified assumption, the number of pairs needed for achieving statistical significance was calculated to be approximately 60-80. A randomized controlled trial is under way to secure enough patients, sustain compliance/adherence and minimize dropouts. PMID- 12578907 TI - The FGF receptor uses the endocannabinoid signaling system to couple to an axonal growth response. AB - A key role for DAG lipase activity in the control of axonal growth and guidance in vitro and in vivo has been established. For example, DAG lipase activity is required for FGF-stimulated calcium influx into neuronal growth cones, and this response is both necessary and sufficient for an axonal growth response. The mechanism that couples the hydrolysis of DAG to the calcium response is not known. The initial hydrolysis of DAG at the sn-1 position (by DAG lipase) will generate 2-arachidonylglycerol, and this molecule is well established as an endogenous cannabinoid receptor agonist in the brain. In the present paper, we show that in rat cerebellar granule neurons, CB1 cannabinoid receptor antagonists inhibit axonal growth responses stimulated by N-cadherin and FGF2. Furthermore, three CB1 receptor agonists mimic the N-cadherin/FGF2 response at a step downstream from FGF receptor activation, but upstream from calcium influx into cells. In contrast, we could find no evidence for the CB1 receptor coupling the TrkB neurotrophin receptor to an axonal growth response in the same neurons. The observation that the CB1 receptor can couple the activated FGF receptor to an axonal growth response raises novel therapeutic opportunities. PMID- 12578908 TI - Substrate-specific function of the translocon-associated protein complex during translocation across the ER membrane. AB - Although the transport of model proteins across the mammalian ER can be reconstituted with purified Sec61p complex, TRAM, and signal recognition particle receptor, some substrates, such as the prion protein (PrP), are inefficiently or improperly translocated using only these components. Here, we purify a factor needed for proper translocation of PrP and identify it as the translocon associated protein (TRAP) complex. Surprisingly, TRAP also stimulates vectorial transport of many, but not all, other substrates in a manner influenced by their signal sequences. Comparative analyses of several natural signal sequences suggest that a dependence on TRAP for translocation is not due to any single physical parameter, such as hydrophobicity of the signal sequence. Instead, a functional property of the signal, efficiency of its post-targeting role in initiating substrate translocation, correlates inversely with TRAP dependence. Thus, maximal translocation independent of TRAP can only be achieved with a signal sequence, such as the one from prolactin, whose strong interaction with the translocon mediates translocon gating shortly after targeting. These results identify the TRAP complex as a functional component of the translocon and demonstrate that it acts in a substrate-specific manner to facilitate the initiation of protein translocation. PMID- 12578910 TI - Cyclic AMP induces integrin-mediated cell adhesion through Epac and Rap1 upon stimulation of the beta 2-adrenergic receptor. AB - cAMP controls many cellular processes mainly through the activation of protein kinase A (PKA). However, more recently PKA-independent pathways have been established through the exchange protein directly activated by cAMP (Epac), a guanine nucleotide exchange factor for the small GTPases Rap1 and Rap2. In this report, we show that cAMP can induce integrin-mediated cell adhesion through Epac and Rap1. Indeed, when Ovcar3 cells were treated with cAMP, cells adhered more rapidly to fibronectin. This cAMP effect was insensitive to the PKA inhibitor H 89. A similar increase was observed when the cells were transfected with Epac. Both the cAMP effect and the Epac effect on cell adhesion were abolished by the expression of Rap1-GTPase-activating protein, indicating the involvement of Rap1 in the signaling pathway. Importantly, a recently characterized cAMP analogue, 8 (4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate, which specifically activates Epac but not PKA, induced Rap-dependent cell adhesion. Finally, we demonstrate that external stimuli of cAMP signaling, i.e., isoproterenol, which activates the G alpha s-coupled beta 2-adrenergic receptor can induce integrin-mediated cell adhesion through the Epac-Rap1 pathway. From these results we conclude that cAMP mediates receptor-induced integrin-mediated cell adhesion to fibronectin through the Epac-Rap1 signaling pathway. PMID- 12578909 TI - Targeting of U4/U6 small nuclear RNP assembly factor SART3/p110 to Cajal bodies. AB - The spliceosomal small nuclear RNAs (snRNAs) are distributed throughout the nucleoplasm and concentrated in nuclear inclusions termed Cajal bodies (CBs). A role for CBs in the metabolism of snRNPs has been proposed but is not well understood. The SART3/p110 protein interacts transiently with the U6 and U4/U6 snRNPs and promotes the reassembly of U4/U6 snRNPs after splicing in vitro. Here we report that SART3/p110 is enriched in CBs but not in gems or residual CBs lacking coilin. The U6 snRNP Sm-like (LSm) proteins, also involved in U4/U6 snRNP assembly, were localized to CBs as well. The levels of SART3/p110 and LSm proteins in CBs were reduced upon treatment with the transcription inhibitor alpha-amanitin, suggesting that CB localization reflects active processes dependent on transcription/splicing. The NH2-terminal HAT domain of SART3/p110 was necessary and sufficient for specific protein targeting to CBs. Overexpression of truncation mutants containing the HAT domain had dominant negative effects on U6 snRNP localization to CBs, indicating that endogenous SART3/p110 plays a role in targeting the U6 snRNP to CBs. We propose that U4 and U6 snRNPs accumulate in CBs for the purpose of assembly into U4/U6 snRNPs by SART3/p110. PMID- 12578911 TI - A tumor-associated beta 1 integrin mutation that abrogates epithelial differentiation control. AB - SCC4 human keratinocytes are derived from a squamous cell carcinoma of the tongue and undergo very little spontaneous differentiation. Introduction of a wild-type beta 1 integrin subunit into SCC4 cells stimulates differentiation, suggesting either that the cells have a defect in the integrin signaling pathways that control differentiation or that the beta1 subunit itself is defective. Here we describe a heterozygous mutation in the SCC4 beta 1 subunit. The mutation, T188I, maps to the I-like domain. It results in constitutive activation of ligand binding, irrespective of the partner alpha subunit, in solid phase assays with recombinant protein and in living cells. The mutation promotes cell spreading, but not proliferation, motility, or invasiveness. It results in sustained activation of Erk MAPK independent of cell spreading. When introduced into SCC4 keratinocytes, the wild-type beta1 integrin stimulates differentiation, whereas the mutant is inactive. Activation of beta 1 integrins in normal keratinocytes also suppresses differentiation. These results establish, for the first time, mutation as a mechanism by which integrins can contribute to neoplasia, because the degree of differentiation in epithelial cancers is inversely correlated with prognosis. They also provide new insights into how integrins regulate keratinocyte differentiation. PMID- 12578913 TI - Agonists vs levodopa in PD: the thrilla of whitha. PMID- 12578912 TI - Regulation of the formation of osteoclastic actin rings by proline-rich tyrosine kinase 2 interacting with gelsolin. AB - Osteoclast activation is important for bone remodeling and is altered in multiple bone disorders. This process requires cell adhesion and extensive actin cytoskeletal reorganization. Proline-rich tyrosine kinase 2 (PYK2), a major cell adhesion-activated tyrosine kinase in osteoclasts, plays an important role in regulating this event. The mechanisms by which PYK2 regulates actin cytoskeletal organization and osteoclastic activation remain largely unknown. In this paper, we provide evidence that PYK2 directly interacts with gelsolin, an actin binding, severing, and capping protein essential for osteoclastic actin cytoskeletal organization. The interaction is mediated via the focal adhesion-targeting domain of PYK2 and an LD motif in gelsolin's COOH terminus. PYK2 phosphorylates gelsolin at tyrosine residues and regulates gelsolin bioactivity, including decreasing gelsolin binding to actin monomer and increasing gelsolin binding to phosphatidylinositol lipids. In addition, PYK2 increases actin polymerization at the fibroblastic cell periphery. Finally, PYK2 interacts with gelsolin in osteoclasts, where PYK2 activation is required for the formation of actin rings. Together, our results suggest that PYK2 is a regulator of gelsolin, revealing a novel PYK2-gelsolin pathway in regulating actin cytoskeletal organization in multiple cells, including osteoclasts. PMID- 12578914 TI - Mortality in epilepsy: searching for clues in populations and patients. PMID- 12578915 TI - Preventing stroke-related seizures: when should anticonvulsant drugs be started? PMID- 12578916 TI - Practice parameter: evaluation of the child with global developmental delay: report of the Quality Standards Subcommittee of the American Academy of Neurology and The Practice Committee of the Child Neurology Society. AB - OBJECTIVE: To make evidence-based recommendations concerning the evaluation of the child with a nonprogressive global developmental delay. METHODS: Relevant literature was reviewed, abstracted, and classified. Recommendations were based on a four-tiered scheme of evidence classification. RESULTS: Global developmental delay is common and affects 1% to 3% of children. Given yields of about 1%, routine metabolic screening is not indicated in the initial evaluation of a child with global developmental delay. Because of the higher yield (3.5% to 10%), even in the absence of dysmorphic features or features suggestive of a specific syndrome, routine cytogenetic studies and molecular testing for the fragile X mutation are recommended. The diagnosis of Rett syndrome should be considered in girls with unexplained moderate to severe developmental delay. Additional genetic studies (e.g., subtelomeric chromosomal rearrangements) may also be considered in selected children. Evaluation of serum lead levels should be restricted to those children with identifiable risk factors for excessive lead exposure. Thyroid studies need not be undertaken (unless clinically indicated) if the child underwent newborn screening. An EEG is not recommended as part of the initial evaluation unless there are historical features suggestive of epilepsy or a specific epileptic syndrome. Routine neuroimaging, with MRI preferred to CT, is recommended particularly if abnormalities are found on physical examination. Because of the increased incidence of visual and auditory impairments, children with global developmental delay may undergo appropriate visual and audiometric assessment at the time of diagnosis. CONCLUSIONS: A specific etiology can be determined in the majority of children with global developmental delay. Certain routine screening tests are indicated and depending on history and examination findings, additional specific testing may be performed. PMID- 12578917 TI - Remote symptomatic epilepsy: does seizure severity increase mortality? AB - OBJECTIVE: To investigate the excess mortality due to remote symptomatic epilepsy, taking account of frequency and type of seizures. METHODS: The authors compared mortality in persons with (n = 8,156) and without (n = 72,526) history of epilepsy in a 1988 to 1999 California population of persons with mild developmental disabilities. Subjects had traumatic brain injury, cerebral palsy, Down syndrome, autism, or no identifiable condition. There were 506,204 person years of data, with 1,523 deaths. Excess death rates and standardized mortality ratios were computed for the persons in the study with epilepsy, relative to those in the study without epilepsy. Controlled comparisons were made using logistic regression on person-years. RESULTS: Compared to subjects with no epilepsy, the excess mortality was six (deaths per 1,000 persons per year) for persons with a recent (<12 months) history of status epilepticus, five for a recent history of generalized tonic-clonic seizure, three for a recent history of nonconvulsive seizures, and less than one for a history of epilepsy but no recent events. Proportion in remission and excess mortality showed no change over the 12 year study period. CONCLUSIONS: Persistent seizures are associated with increased mortality in remote symptomatic epilepsy. Mortality is highest among individuals with status epilepticus or generalized convulsions. PMID- 12578918 TI - Early and late seizures after cryptogenic ischemic stroke in young adults. AB - OBJECTIVES: To assess the incidence and predictive factors of early and late seizures after ischemic stroke in young adults. METHODS: A total of 581 patients (aged 18 to 55 years) with recent cryptogenic ischemic stroke were prospectively enrolled at 30 neurology departments and followed for 37.8 +/- 9.7 months. Early seizures (occurring within 7 days of stroke) were assessed by chart review and late seizures were prospectively recorded at each follow-up visit. Clinical and brain imaging findings were reviewed by two neurologists and two neuroradiologists who were blinded to the occurrence of seizures. RESULTS: Fourteen of the 581 patients (2.4%) developed early seizures, 71% of which occurred within the first 24 hours. Rankin scale >or=3 (odds ratio [OR] 3.9, 95% CI 1.2 to 12.7) and cortical involvement (OR 7.7, 95% CI 1.0 to 61.1) were independently associated with early seizures. Late seizures occurred only in patients with hemispheric stroke (n = 20). The risk of first late seizure was 3.1% (95% CI 1.4 to 4.8) within 1 year and 5.5% (95% CI 3.1 to 7.9) within 3 years. The mean delay between stroke and first late seizure was 12.9 months (0.3 to 33.9). Late seizures were associated with early seizure (hazard ratio [HR] 5.1, 95% CI 1.8 to 14.8), cortical signs (HR 4.5, 95% CI 1.6 to 13.1), and size of infarct superior to one-half hemisphere (HR 9.7, 95% CI 3.1 to 30.8). Eleven of the 20 patients with late seizure experienced recurrences (multiple in eight) on antiepileptic drug treatment. Most of them were seizure free at the end of the follow-up. CONCLUSION: Epilepsy is rarely a major problem in young cryptogenic ischemic stroke survivors. Early seizures are associated with stroke disability and cortical involvement. Early seizures, cortical signs, and large infarct are independent risk factors for late seizures. PMID- 12578919 TI - Hippocampal atrophy and T2-weighted signal changes in familial mesial temporal lobe epilepsy. AB - OBJECTIVE: To correlate the clinical phenotype with hippocampal volumes (HcVs) and signal changes in patients with familial mesial temporal lobe epilepsy (FMTLE). METHODS: FMTLE was defined when at least two first-degree relatives in a family had a clinical-EEG diagnosis of MTLE. Hippocampal formation measurements were performed using 1- to 3-mm coronal T1-weighted MRIs. The presence of hyperintense T2 signal was evaluated by visual analysis. For statistical analyses, analysis of variance, chi(2) test, and regression analysis were used. RESULTS: A total of 142 patients from 45 unrelated families were studied: 113 individuals with MTLE (80 with good seizure control) and 29 family members with other seizure types. There were 99 patients (69.7%) with hippocampal atrophy (HA). Sixty-seven of the 99 patients with HA also had a hyperintense T2 signal. Hyperintense T2 signal was associated with more severe HA (p = 0.04). Patients with refractory FMTLE had more frequent HA (p = 0.03) and hyperintense T2 signal (p = 0.004) and more severe atrophy (p < 0.0001). Duration of epilepsy correlated with HcV asymmetry index (r(2) = 0.12, p = 0.00008) and with the more atrophic hippocampi but not with contralateral hippocampi. CONCLUSION: In familial mesial temporal lobe epilepsy, seizure severity is variable in affected individuals. Hippocampal atrophy was present in 70% of these patients and 69% of these had an associated hyperintense T2 signal. Although hippocampal atrophy associated with abnormal T2 signal was more frequent and more severe in patients with poor seizure control, it was also frequent in affected individuals across families. These observations suggest that one or more genes resulting in familial mesial temporal lobe epilepsy predisposes both to the clinical features of mesial temporal lobe epilepsy and to the development of hippocampal sclerosis. PMID- 12578920 TI - Phenotypic features of familial febrile seizures: case-control study. AB - OBJECTIVE: To identify phenotypic features of febrile seizures that can be used to reduce heterogeneity and thereby increase power in linkage analysis. BACKGROUND: Despite exciting discoveries in several rare pedigrees, the genetic basis of common febrile seizures remains a mystery. The major drawback of studying common febrile seizure disorder is etiologic and genetic heterogeneity. A linkage sample must therefore be classified a priori on phenotypic criteria likely to reflect genetically homogeneous subgroups. METHODS: Eighty-three cases (children with one or more febrile seizure plus first-degree family history of febrile seizures) and 101 controls (children with one or more febrile seizure but no first-degree family history of febrile seizures) were compared for association of phenotypic features in an unmatched case-control design. Odds ratios were calculated using univariate and multivariate methods. RESULTS: Recurrent febrile seizures was the only phenotypic feature significantly associated with first degree family history of febrile seizures (OR 2.1, 95% CI 1.15 to 3.88). First degree family history and later occurrence of afebrile seizures (OR 3.47, 95% CI 0.94 to 12.78) were independently associated with recurrent febrile seizures. Complex features did not show familial aggregation. CONCLUSIONS: The authors suggest recurrent and afebrile seizures as criteria on which to subgroup a linkage sample. These subgroups will not be evident at the time of the initial febrile seizure. Meticulous and prospective collection of phenotypic and family data are recommended. PMID- 12578921 TI - Spike-triggered fMRI in reading epilepsy: involvement of left frontal cortex working memory area. AB - OBJECTIVE: To determine the origin of epileptiform activity in reading epilepsy (RE) and the association between these regions and regions activated by reading, and to assess brain morphometry in these areas. METHODS: In two subjects with RE, EEG was recorded inside the three tesla MRI while subjects read silently. Spike triggered fMRI images were compared to baseline. In a second fMRI study, 30 seconds of silent reading was compared to visual fixation. Morphometry of these areas was assessed using curvilinear surface reconstruction. Left central sulcal patterns in three subjects with RE were compared to three subjects with idiopathic generalized epilepsy (IGE) and 12 normal controls. RESULTS: One subject with RE showed spike-related activity (17 spikes) in the left precentral gyrus, and bilaterally in the central sulcus and globus pallidus. The other showed no definite activation owing to low spike numbers (4 spikes). In both subjects, the block reading task recruited normal visual and language areas including the left posterior middle frontal gyrus. Two subjects with RE showed an unusual gyrus branching anteriorly off the left central sulcus. A similar sulcal pattern was seen in none of the subjects with IGE and only 1 of 12 controls. CONCLUSION: Spike activity overlapped with reading activity in the left middle frontal gyrus, a structure recruited during working memory cognitive tasks. The authors postulate that, because of a local structural anomaly, the spikes of reading epilepsy spread from working memory areas into adjacent motor cortex, activating a cortical subcortical circuit. PMID- 12578922 TI - Rasmussen's encephalitis: early characteristics allow diagnosis. AB - OBJECTIVE: To identify early manifestations of Rasmussen encephalitis (RE) that can prompt early and reasonably secure diagnosis, allowing medical or surgical therapies at an early stage when they may be more effective in slowing the disease. METHODS: The authors studied 12 patients with clinical and neuropathologic diagnosis of RE, followed from disease onset, assessing clinical history, imaging, and EEG and focusing on early characteristics. Anti-GluR3 antibody assays were also considered in 11 patients. RESULTS: By 4 months from first symptoms, all cases had 1) refractory focal seizures with a predominant motor component, 2) slow focal activity on EEG contralateral to the motor manifestations, and 3) focal contralateral white matter hyperintensity with insular cortical atrophy on neuroimaging. Less constant or later findings were epilepsia partialis continua, oligoclonal bands, and serum anti-GluR3 antibodies. CONCLUSIONS: The association of partial seizures with focal EEG and neuroimaging changes allows a tentative diagnosis of RE 4 to 6 months after first symptoms. PMID- 12578923 TI - Impaired facial emotion recognition in early-onset right mesial temporal lobe epilepsy. AB - BACKGROUND: Anteromedial temporal lobe regions, particularly the amygdala, participate in the recognition of emotions from facial expressions. The authors studied the ability of facial emotion recognition (ER) in subjects with symptomatic epilepsy, evaluating whether mesial temporal lobe damage is related to an impairment in the recognition of specific emotions and whether the onset of seizures in a critical period of life could prevent the development of ER. METHODS: Groups included patients with temporal lobe epilepsy (TLE) with MRI evidence of mesial temporal sclerosis (MTS) (n = 33); patients with TLE with MRI evidence of temporal lobe lesions other than MTS (n = 30); and patients with extratemporal epilepsy (n = 33). Healthy volunteers (n = 50) served as controls. ER was tested by matching a facial expression with the name of one of the following basic emotions: happiness, sadness, fear, disgust, and anger. A face matching task was used to control visuoperceptual abilities with face stimuli. RESULTS: No subject showed deficits in the face-matching task. ER was impaired in patients with right MTS, especially for fearful faces. Patients presenting left MTS, right or left temporal lobe lesions other than MTS, or extratemporal seizure foci showed ER performances similar to controls. In all subjects with right TLE, the degree of emotion recognition impairment was related to age at first seizure (febrile or afebrile) and age at epilepsy onset. CONCLUSIONS: Early-onset right sided mesial temporal lobe epilepsy is the key substrate determining a severe deficit in recognizing emotional facial expressions, especially fear. PMID- 12578924 TI - Hypophonia in Parkinson's disease: neural correlates of voice treatment revealed by PET. AB - OBJECTIVE: To investigate the neural correlates of hypophonia in individuals with idiopathic PD (IPD) before and after voice treatment with the Lee Silverman Voice Treatment method (VT) using (15)O-H(2)O PET. METHODS: Regional cerebral blood flow (rCBF) changes associated with overt speech-motor tasks relative to the resting state were measured in the IPD subjects before and after VT, and in a group of healthy control volunteers. RESULTS: Behavioral measures of voice loudness significantly improved following treatment. Before VT, patients had strong speech-related activations in motor and premotor cortex (M1-mouth, supplementary motor cortex [SMA], and inferior lateral premotor cortex [ILPm]), which were significantly reduced post-VT. Similar to the post-treatment session, premotor activations were absent (SMA) or below statistical threshold (M1-mouth) in the healthy control group. In addition, following VT treatment, significant right-sided activations were present in anterior insular cortex, caudate head, putamen, and dorsolateral prefrontal cortex (DLPFC). Finally, the VT-induced neural changes were not present with transient experimenter-cued increases of loudness in VT-untreated patients. CONCLUSIONS: Effective improvement of IPD hypophonia following voice treatment with VT was accompanied by a reduction of cortical motor-premotor activations, resembling the functional pattern observed in healthy volunteers and suggesting normalization, and additional recruitment of right anterior insula, caudate head, putamen, and DLPFC. This treatment-dependent functional reorganization suggests a shift from an abnormally effortful (premotor cortex) to a more automatic (basal ganglia, anterior insula) implementation of speech-motor actions. PMID- 12578925 TI - Patterns of health care utilization for migraine in England and in the United States. AB - OBJECTIVE: To assess patterns of medical consultation, diagnosis, and medication use in representative samples of adults with migraine in England and the United States. METHODS: Validated computer-assisted telephone interviews were conducted in the United Kingdom (n = 4,007) and the United States (n = 4,376). Individuals who reported six or more headaches per year meeting the criteria for migraine were interviewed. RESULTS: Patients with migraine in the United Kingdom were more likely to have consulted a doctor for headache at least once in their lifetime (86% vs 69%, p < 0.0001), but also were more likely to have lapsed from medical care (37% vs 21%, p < 0.001). In the United States, patients with migraine who had consulted made more office visits for headache and were more likely to see a specialist. In the United States, but not in the United Kingdom, women with migraine were more likely than men to consult doctors for headache. Patients with migraine in the United Kingdom were more likely to receive a medical diagnosis of migraine (UK 67%, US 56%; p < 0.05). Patterns of medication use were similar in both countries, with most people treating with over-the-counter (OTC) medications. Substantial disability occurred in a high proportion of those who never consulted (UK 60%, US 68%), never received a correct medical diagnosis (UK 64%, US 77%), and treated only with OTC medication (UK 72%, US 70%). CONCLUSION: Medically unrecognized migraine remains an important health problem both in the United States and the United Kingdom. Furthermore, there may be barriers to consultation for men in the United States that do not operate in the United Kingdom. PMID- 12578927 TI - A Bayesian argument against rigid cut-offs in electrodiagnosis of median neuropathy at the wrist. AB - BACKGROUND: Nerve conduction (NC) tests, using rigid cut-offs separating normal from abnormal test values, are commonly used to confirm median neuropathy at the wrist (MNW). The authors studied patients with clinically defined mild MNW and a normal median distal motor latency to determine 1) how much sensory or mixed NC test results increase (or decrease) the probability of MNW and 2) the NC test values required to confirm (or exclude) MNW for the range of pretest probabilities of MNW. METHODS: Palmar, digit 4 (D4), and digit 2 (D2) median NC tests were reviewed in 125 hands with mild carpal tunnel syndrome (CTS) and 100 control hands with musculoskeletal pain. Receiver operating characteristic curves and interval likelihood ratios were plotted for the three tests. Using Bayes theorem, post-test probability of MNW was then determined for the range of pretest probabilities and NC test values. RESULTS: Receiver operating characteristic curves showed that for a set specificity of 97%, palmar and D4 studies had higher electrodiagnostic utility than D2 studies with cut-off test values (sensitivities of 0.3 msec, 64.0%; 0.4 msec, 71.2%; and 50 m/sec, 44.8%). However, Bayesian analysis showed that to confirm MNW more conservative cut-off values (palmar 0.5 msec, D4 0.7 msec, D2 44 m/sec) were required for pretest probabilities or=75%. Conversely, normal test values could exclude MNW only for pretest probabilities <25%. CONCLUSIONS: For a given NC test value, post-test probability of MNW can be determined from the estimated pretest probability (derived from clinical data), interval likelihood ratios, and Bayes theorem. Use of rigid cut-off values to confirm MNW is problematic, because more conservative cut-offs are required for low pretest probability. Conversely, NC tests with sensitivity <95% cannot exclude MNW when pretest probability is high. PMID- 12578926 TI - Impaired financial abilities in mild cognitive impairment: a direct assessment approach. AB - OBJECTIVES: To assess financial capacity in patients with mild cognitive impairment (MCI) using a standardized psychometric capacity measure. METHODS: Participants were 21 cognitively normal older controls, 21 patients with amnestic MCI, and 22 patients with mild AD. The Financial Capacity Instrument (FCI), a psychometric capacity measure consisting of 18 financial ability tests (tasks), 9 domains (activities), and 2 total scores, was administered to participants along with a battery of neuropsychological tests sensitive to dementia. Group differences were examined on the neuropsychological and financial capacity variables. RESULTS: Relative to controls, the MCI group demonstrated impairments in episodic memory, and also semantic knowledge, executive function, written arithmetic, and spatial attention. MCI participants demonstrated impairments in FCI domains of conceptual knowledge, cash transactions, bank statement management, and bill payment, and in overall financial capacity. The control and MCI groups performed significantly better than patients with AD on most financial capacity and cognitive measures. CONCLUSIONS: On direct assessment, patients with amnestic MCI as a group demonstrate impairments across a range of financial abilities. These impairments are mild and may only apply to a subset of patients with MCI. However, existing diagnostic criteria for MCI should be applied flexibly to include mild impairments in higher order activities of daily life such as financial capacity. PMID- 12578928 TI - Quantitative sensory testing cannot differentiate simulated sensory loss from sensory neuropathy. AB - OBJECTIVE: To differentiate the quantitative sensory testing (QST) results of subjects simulating small and large fiber sensory loss from those of normal subjects and subjects with sensory peripheral neuropathy. BACKGROUND: QST is used to measure sensory thresholds in clinical, epidemiologic, and research studies. It is not known whether there are objective test results that characterize the subject seeking to deceive the examiner. METHODS: The Computer Aided Sensory Examination IV 4, 2, and 1 stepping algorithm was used to determine vibration and cold perception in nine naive subjects. Subjects were asked to simulate sensory loss (on two occasions) and to respond normally on one occasion. Test results were compared to those of subjects with diabetic sensory neuropathy. Each QST trial was performed three times. RESULTS: Reproducibility, measured by the intraclass correlation coefficient, was similar in all groups for the vibration perception test (simulation 1: 0.68 [95% CI 0.31, 0.91], simulation 2: 0.82 [95% CI 0.54, 0.95], normal response: 0.77 [95% CI 0.47, 0.94], and subjects with peripheral neuropathy: 0.76 [95% CI 0.18, 0.95]) and the cold perception test (simulation 1: 0.53 [95% CI 0.12, 0.85], simulation 2: 0.82 [95% CI 0.55, 0.95], normal subjects: 0.67 [95% CI 0.30, 0.90] and subjects with peripheral neuropathy: 0.88 [95% CI 0.57, 0.97]), all just noticeable difference units. There were no differences between performance characteristics in the two simulation trials. Responses to null stimuli did not differentiate between groups. CONCLUSION: Test performance characteristics do not permit discrimination among subjects simulating sensory loss, subjects with normal responses, and subjects with peripheral neuropathy. PMID- 12578929 TI - Sex differences in carotid endarterectomy utilization and 30-day postoperative mortality. AB - OBJECTIVE: To study trends, and sex and regional differences in utilization of the carotid endarterectomy (CEA) procedure and 30-day postoperative mortality from 1991 to 1999. METHODS: Retrospective analysis of fee-for-service claims and mortality data for Medicare beneficiaries aged 65 years and older in the United States. RESULTS: The male and female CEA rates and 30-day mortality increased with age up to the age of 79 years. From 1991 to 1995, the age-adjusted male and female CEA rates increased 72% from 26.6 and 14.2 procedures per 10,000 beneficiaries. Thereafter, the CEA rates slightly decreased except for the 80 years and older age group, which increased through 1999. In each year from 1991 to 1999, the age-adjusted male CEA rates were approximately 1.9 times higher than the corresponding female rates. From 1991 to 1998, the age-adjusted male and female 30-day mortality decreased 29.3% and 46.4% from 19.2 and 18.1 deaths per 1,000 procedures. From 1992 to 1997, except 1994, 30-day mortality was higher in men than in women. This sex difference was not present in the 65 to 69 years age group. There were small differences in CEA rates between two of the four regions of the United States in 3 of the 9 years. CONCLUSIONS: Increasing CEA rates with decreasing postoperative mortality suggest that CEA may have been more frequently performed on low-risk patients. The apparent sex differences in CEA rates may not be true differences. PMID- 12578930 TI - Mild cognitive impairment in the oldest old. AB - BACKGROUND: No data exist on whether the syndrome of amnestic mild cognitive impairment occurs in the oldest old, or if the relationships for functional status and neuropsychometric performance based on clinical diagnosis hold true in this age group. DESIGN/METHODS: The authors performed comprehensive neurologic evaluations, neuropsychometric testing, and functional assessments on a sample of 90- to 100-year-old residents of Rochester, MN. Subjects were diagnosed as normal or with amnestic mild cognitive impairment (MCI) or dementia according to well accepted criteria. Data on the following measures were collected and analyzed: Record of Independent Living (ROIL), Mini-Mental State Examination (MMSE), Dementia Rating Scale (DRS), Trailmaking Test (TMT), and modified version of the Free and Cued Selective Reminding Test (FCSRT). RESULTS: Data on 111 subjects (56 normal, 13 MCI, and 42 dementia) were analyzed. On the ROIL, functional capacity to carry out activities of daily living was worse for patients with dementia compared to patients with MCI and normal subjects, but did not differ between MCI and normal subjects. Scores on the MMSE, DRS, and TMT-A were worse in the dementia group compared to the normal group, and in the dementia group compared to MCI, but scores on these measures for normal subjects compared to patients with MCI were not different. Scores on the FCSRT and memory subtest of the DRS showed differences among all three groups. CONCLUSION: In spite of the advanced age of the cohort, the relationship between cognitive and functional performance and clinical diagnosis follows patterns previously described in younger samples of normal subjects, subjects with amnestic mild cognitive impairment, and subjects with dementia. PMID- 12578932 TI - Ecological implications of ideomotor apraxia: evidence from physical activities of daily living. AB - OBJECTIVE: To learn if ideomotor apraxia (IMA) adversely influences skilled acts in the environment and interferes with independent functioning after stroke. METHODS: The relationship between IMA severity, based on scores from a verbal gesture-to-command (pantomime) task, and the dependency score, as defined by increased caregiver assistance on the Physical Self-Maintenance Scale (PSMS), was investigated in 10 unilateral left hemisphere-damaged stroke patients and 10 matched control subjects. RESULTS: There was a significant relationship between apraxia severity and dependency in physical functioning (PSMS). Impairment on the PSMS in the patients with IMA could not be accounted for based on overall cognitive impairment, poststroke depression, content-conceptual errors, elementary motor impairment, lesion size, or stroke-test interval. Analysis of categories composing the PSMS revealed that the patients with apraxia had increased dependency in grooming, bathing, and toileting relative to age-matched control subjects. CONCLUSIONS: These findings emphasize the ecological implications of apraxia and the need for rehabilitation strategies to improve the execution and efficiency of coordinated skilled movements in stroke patients with left hemisphere damage. PMID- 12578933 TI - Split spinal cord (diastematomyelia). PMID- 12578931 TI - Atrophy rates of entorhinal cortex in AD and normal aging. AB - OBJECTIVES: To explore the atrophy rate of entorhinal cortex (ERC) in AD and normal aging and assess the value of rate measurement of ERC atrophy for classifying subjects with AD from cognitively normal (CN) control subjects. METHODS: Twenty-one AD patients and 23 CN subjects had MRI scans and clinical evaluations twice within 1.8 +/- 0.6 years. ERC volumes were manually measured on volumetric T1-weighted MR images. RESULTS: Patients with AD had a greater annual percentage volume change of ERC than CN subjects on both sides (left: 6.8 +/- 4.3%/year for AD vs 1.4 +/- 2.5%/year for CN [F(1,42) = 25.6, p < 0.001]; right: 6.3 +/- 3.3%/year for AD vs 1.4 +/- 2.3%/year for CN [F(1,42) = 25.6, p < 0.001]). Furthermore, increased ERC atrophy rate was correlated (r = -0.56, p = 0.01) with decreased memory performance in AD. CN subjects had on average annual ERC atrophy rates greater than zero (p < 0.01). Baseline volume of ERC predicted atrophy rate of ERC (left: r = -0.53, p < 0.01; right: r = -0.42, p < 0.05) in CN subjects but not in AD subjects. Using ERC baseline volumes alone resulted in 77% overall correct classification (p < 0.01) between AD and CN subjects, with 76% sensitivity and 78% specificity and an area under receiver operator characteristic (ROC) curve of 0.83. Adding annual atrophy rate of ERC to the model accounted for most of the variance (p < 0.01), diminishing contributions from baseline volume and yielding 82% overall classification, with 76% sensitivity and 86% specificity and an area under the ROC curve of 0.93. CONCLUSION: ERC volume loss over time may be a better indicator for AD than cross sectional measurements. PMID- 12578934 TI - Evidence of cardiac ischemia during seizures in drug refractory epilepsy patients. AB - The authors investigated whether patients with drug refractory epilepsy have cardiovascular abnormalities that might be related to a high frequency of sudden death. Twenty-three subjects underwent comprehensive cardiovascular evaluations before and during video-EEG monitoring. ST-segment depression occurred in 40% and was associated with a higher maximum heart rate during seizures. These data suggest that cardiac ischemia may occur in these patients. PMID- 12578935 TI - Onset of epilepsy at the time of menarche. AB - The authors compared the frequency of epilepsy onset in perimenarche with epilepsy onset in other childhood periods in 94 postmenarchial patients aged <55 years. Seizure onset was higher for the 12 to 15 year age bracket than for other ages and clustered around menarche. Epilepsy began during the year of menarche in 17% of patients vs 5.5% expected (p < 0.001), and during +/-2 years of menarche in 38% patients vs 22% expected (p < 0.001). Seizures worsened during perimenarche in 29% of girls with pre-existing epilepsy. Perimenarche may be a risk for the development and worsening of epilepsy. PMID- 12578936 TI - Improvement of voicing in patients with Parkinson's disease by speech therapy. AB - Speech therapy in PD patients, focusing on an increase of phonatory-respiratory effort, has adverse effects because it raises vocal pitch and laryngeal muscle tension. The authors' approach, the Pitch Limiting Voice Treatment (PLVT), increases loudness but at the same time sets vocal pitch at a better level. In this study, the Lee Silverman Voice Treatment ("think loud, think shout") and PLVT ("speak loud and low") are compared. Both treatments produce the same increase in loudness, but PLVT limits an increase in vocal pitch and prevents a strained or pressed voicing. PMID- 12578937 TI - Creatine monohydrate in DM2/PROMM: a double-blind placebo-controlled clinical study. Proximal myotonic myopathy. AB - The efficacy and safety of creatine monohydrate (Cr) in patients with myotonic dystrophy type 2/proximal myotonic myopathy were studied in a small placebo controlled double-blind trial. Twenty patients received either Cr or placebo for 3 months. After 3 months, there were no significant differences of muscle strength as assessed by hand-held dynamometry, testing of maximum grip strength, Medical Research Council scoring, and the Neuromuscular Symptom Score between the two groups. Some measures indicated trends toward mild improvement with Cr. Myalgia improved in two patients. PMID- 12578938 TI - Benefit of IVIG for long-standing ataxic sensory neuronopathy with Sjogren's syndrome. IV immunoglobulin. AB - Ataxic sensory neuronopathy with Sjogren's syndrome is a devastating neurologic complication for which there is no established treatment. IV immunoglobulin (IVIG) was given to five patients with severe disabilities for an average of 12 years. Four patients showed remarkable improvement, two of whom responded after the first course. The authors conclude that IVIG is safe and effective to treat even chronically debilitated patients who have the disease, presumably because it ameliorates smoldering inflammation. PMID- 12578939 TI - Deafness and CMT disease associated with a novel four amino acid deletion in the PMP22 gene. AB - The molecular basis for the clinically distinct entity of deafness with Charcot Marie-Tooth disease has not been established with certainty. The authors report deafness associated with a demyelinating neuropathy in three individuals of a family in whom a novel four-amino acid deletion in the PMP22 gene was identified. The data and review of literature suggest that in the PMP22 gene, some point mutations and small deletions in the transmembrane domain that are in close proximity to the extracellular component of the protein result in this clinically distinct entity. PMID- 12578940 TI - Obstacles in the diagnosis and treatment of syphilitic amyotrophy. AB - The authors describe a case of syphilitic amyotrophy in a 37-year-old man presenting with subacute progressive painless weakness in the right arm. Syphilitic amyotrophy is still an existing differential diagnosis of painless and progressive weakness. The authors explore potential obstacles in the diagnostic workup and finding of adequate therapy. PMID- 12578941 TI - Asymptomatic microbleeds as a risk factor for aspirin-associated intracerebral hemorrhages. AB - The authors measured the presence and extent of asymptomatic microbleeds on gradient-recalled-echo MRI in 21 aspirin users who developed intracerebral hemorrhage and 21 aspirin users without history of intracerebral hemorrhage. Microbleeds were more frequent (19 vs 7, p < 0.001) and more extensive (mean number of microbleeds 13.3 vs 0.4, p < 0.001) in the intracerebral hemorrhage group than in the control group. Asymptomatic microbleeds may be a risk factor for aspirin-associated intracerebral hemorrhage. PMID- 12578942 TI - Sporadic Creutzfeldt-Jakob disease with MM1-type prion protein and plaques. AB - The authors report a 75-year-old woman with atypical sporadic Creutzfeldt-Jakob disease (CJD) characterized by MM1-type prion protein (PrP) (methionine homozygosity at codon 129 in the PrP gene and type-1 protease-resistant PrP) and PrP plaques. This patient is the first case of sporadic CJD with plaque-forming MM1-type PrP, suggesting either a shared prion strain with the plaque-forming subset of dural graft-associated CJD or shared host genetic factors that are unrelated to the PrP genotype. PMID- 12578944 TI - Microhemorrhages on gradient echo MRI. PMID- 12578943 TI - Multifocal myelitis in Behcet's disease. PMID- 12578945 TI - Reversal of parkinsonism following liver transplantation. PMID- 12578946 TI - Microvascular decompression for trigeminal neuralgia in patients over 70 years of age. PMID- 12578947 TI - Transient eye and nose pain as an initial symptom of pontine infarction. PMID- 12578948 TI - Electrophysiology of the myoclonus in dementia with Lewy bodies. PMID- 12578949 TI - Ocular flutter in a patient with intracranial hypertension following cerebral venous thrombosis. PMID- 12578950 TI - Clinical features of frontotemporal dementia due to the intronic tau 10(+16) mutation. PMID- 12578951 TI - Blood pressure control in stroke patients. What should the consulting neurologist advise? PMID- 12578952 TI - Homocysteine, MTHFR 677C-->T polymorphism, and risk of ischemic stroke: results of a meta-analysis. PMID- 12578953 TI - Reduced inhibition within primary motor cortex in patients with poststroke focal motor seizures. PMID- 12578956 TI - Retraction: Shamim et Al. Nonsurgical reduction of the interventricular septum in patients with hypertrophic cardiomyopathy. N Engl J Med 2002;347:1326-33. PMID- 12578957 TI - Notice of retraction. PMID- 12578958 TI - Loading of the human 9-1-1 checkpoint complex onto DNA by the checkpoint clamp loader hRad17-replication factor C complex in vitro. AB - The human DNA damage sensors, Rad17-replication factor C (Rad17-RFC) and the Rad9 Rad1-Hus1 (9-1-1) checkpoint complex, are thought to be involved in the early steps of the DNA damage checkpoint response. Rad17-RFC and the 9-1-1 complex have been shown to be structurally similar to the replication factors, RFC clamp loader and proliferating cell nuclear antigen polymerase clamp, respectively. Here, we demonstrate functional similarities between the replication and checkpoint clamp loader/DNA clamp pairs. When all eight subunits of the two checkpoint complexes are coexpressed in insect cells, a stable Rad17-RFC/9-1-1 checkpoint supercomplex forms in vivo and is readily purified. The two individually purified checkpoint complexes also form a supercomplex in vitro, which depends on ATP and is mediated by interactions between Rad17 and Rad9. Rad17-RFC binds to nicked circular, gapped, and primed DNA and recruits the 9-1-1 complex in an ATP-dependent manner. Electron microscopic analyses of the reaction products indicate that the 9-1-1 ring is clamped around the DNA. PMID- 12578959 TI - An alpha-syntrophin-dependent pool of AQP4 in astroglial end-feet confers bidirectional water flow between blood and brain. AB - The water channel AQP4 is concentrated in perivascular and subpial membrane domains of brain astrocytes. These membranes form the interface between the neuropil and extracerebral liquid spaces. AQP4 is anchored at these membranes by its carboxyl terminus to alpha-syntrophin, an adapter protein associated with dystrophin. To test functions of the perivascular AQP4 pool, we studied mice homozygous for targeted disruption of the gene encoding alpha-syntrophin (alpha Syn(-/-)). These animals show a marked loss of AQP4 from perivascular and subpial membranes but no decrease in other membrane domains, as judged by quantitative immunogold electron microscopy. In the basal state, perivascular and subpial astroglial end-feet were swollen in brains of alpha-Syn(-/-) mice compared to WT mice, suggesting reduced clearance of water generated by brain metabolism. When stressed by transient cerebral ischemia, brain edema was attenuated in alpha-Syn( /-) mice, indicative of reduced water influx. Surprisingly, AQP4 was strongly reduced but alpha-syntrophin was retained in perivascular astroglial end-feet in WT mice examined 23 h after transient cerebral ischemia. Thus alpha-syntrophin dependent anchoring of AQP4 is sensitive to ischemia, and loss of AQP4 from this site may retard the dissipation of postischemic brain edema. These studies identify a specific, syntrophin-dependent AQP4 pool that is expressed at distinct membrane domains and which mediates bidirectional transport of water across the brain-blood interface. The anchoring of AQP4 to alpha-syntrophin may be a target for treatment of brain edema, but therapeutic manipulations of AQP4 must consider the bidirectional water flux through this molecule. PMID- 12578960 TI - Identification of frame-shift intermediate mutant cells. AB - Frame-shift mutations at microsatellites occur as a time-dependent function of polymerase errors followed by failure of postreplicational mismatch repair. A cell-culture system was developed that allows identification of intermediate mutant cells that carry the mutation on a single DNA strand after the initial DNA polymerase errors. A plasmid was constructed that contained 13 repeats of a poly(dC-dA).poly(dG-dT) oligonucleotide immediately after the translation initiation codon of the enhanced GFP (EGFP) gene, shifting the EGFP gene out of its proper reading frame. The plasmid was introduced into human mismatch repair deficient (HCT116, hMLH1-mutated) and mismatch repair-proficient (HCT116+chr3, hMLH1 wild type) colorectal cancer cells. After frame-shift mutations occurred that restored the EGFP reading frame, EGFP-expressing cells were detected, and two distinct fluorescent populations, M1 (dim cells) and M2 (bright cells), were identified. M1 cell numbers were stable, whereas M2 cells accumulated over time. In HCT116, single M2 cells gave rise to fluorescent colonies that carried a 2-bp deletion at the (CA)(13) microsatellite. Twenty-eight percent of single M1 cells, however, gave rise to colonies with a mixed fluorescence pattern that carried both (CA)(13) and (CA)(12) microsatellites. It is likely that M1 cells represent intermediate mutants that carry (CA)(13).(GT)(12) heteroduplexes. Although the mutation rate in HCT116 cell clones (6.2 x 10(-4)) was 30 times higher than in HCT116+chr3 (1.9 x 10(-5)), the proportion of M1 cells in culture did not significantly differ between HCT116 (5.87 x 10(-3)) and HCT116+chr3 (4.13 x 10( 3)), indicating that the generation of intermediate mutants is not affected by mismatch-repair proficiency. PMID- 12578961 TI - Operant behavior can be triggered by the position of the rat relative to objects rotating on an inaccessible platform. AB - The present study describes a task testing the ability of rats to trigger operant behavior by their relative spatial position to inaccessible rotating objects. Rats were placed in a Skinner box with a transparent front wall through which they could observe one or two adjacent objects fixed on a slowly rotating arena (d = 1 m) surrounded by an immobile black cylinder. The direction of arena rotation was alternated at a sequence of different time intervals. Rats were reinforced for the first bar-press that was emitted when a radius separating the two adjacent objects or dividing a single object into two halves (pointing radius) entered a 60 degrees sector of its circular trajectory defined with respect to the stationary Skinner box (reward sector). Well trained rats emitted 62.1 +/- 3.6% of responses in a 60 degrees sector preceding the reward sector and in the first 30 degrees of the reward sector. Response rate increased only when the pointing radius was approaching the reward sector, regardless of the time elapsed from the last reward. In the extinction session, when no reward was delivered, rats responded during the whole passage of the pointing radius through the former reward sector and spontaneously decreased responding after the pointing radius left this area. This finding suggests that rats perceived the reward sector as a continuous single region. The same results were obtained when the Skinner box with the rat was orbiting around the immobile scene. It is concluded that rats can recognize and anticipate their position relative to movable objects. PMID- 12578962 TI - Seeds of understanding of plant diversity. PMID- 12578963 TI - A mechanism converting psychosocial stress into mononuclear cell activation. AB - Little is known about the mechanisms converting psychosocial stress into cellular dysfunction. Various genes, up-regulated in atherosclerosis but also by psychosocial stress, are controlled by the transcription factor nuclear factor kappaB (NF-kappaB). Therefore, NF-kappaB is a good candidate to convert psychosocial stress into cellular activation. Volunteers were subjected to a brief laboratory stress test and NF-kappaB activity was determined in peripheral blood mononuclear cells (PBMC), as a window into the body and because PBMC play a role in diseases such as atherosclerosis. In 17 of 19 volunteers, NF-kappaB was rapidly induced during stress exposure, in parallel with elevated levels of catecholamines and cortisol, and returned to basal levels within 60 min. To model this response, mice transgenic for a strictly NF-kappaB-controlled beta-globin transgene were stressed by immobilization. Immobilization resulted in increased beta-globin expression, which could be reduced in the presence of the alpha1 adrenergic inhibitor prazosin. To define the role of adrenergic stimulation in the up-regulation of NF-kappaB, THP-1 cells were induced with physiological amounts of catecholamines for 10 min. Only noradrenaline resulted in a dose- and time-dependent induction of NF-kappaB and NF-kappaB-dependent gene expression, which depended on pertussis-toxin-sensitive G protein-mediated phosphophatidylinositol 3-kinase, Ras/Raf, and mitogen-activated protein kinase activation. Induction was reduced by alpha(1)- and beta-adrenergic inhibitors. Thus, noradrenaline-dependent adrenergic stimulation results in activation of NF kappaB in vitro and in vivo. Activation of NF-kappaB represents a downstream effector for the neuroendocrine response to stressful psychosocial events and links changes in the activity of the neuroendocrine axis to the cellular response. PMID- 12578964 TI - Loss of WAVE-1 causes sensorimotor retardation and reduced learning and memory in mice. AB - The Scar/WAVE family of scaffolding proteins organize molecular networks that relay signals from the GTPase Rac to the actin cytoskeleton. The WAVE-1 isoform is a brain-specific protein expressed in variety of areas including the regions of the hippocampus and the Purkinje cells of the cerebellum. Targeted disruption of the WAVE-1 gene generated mice with reduced anxiety, sensorimotor retardation, and deficits in hippocampal-dependent learning and memory. These sensorimotor and cognitive deficits are analogous to the symptoms of patients with 3p-syndrome mental retardation who are haploinsufficient for WRP/MEGAP, a component of the WAVE-1 signaling network. Thus WAVE-1 is required for normal neural functioning. PMID- 12578965 TI - Mechanosensation through integrins: cells act locally but think globally. PMID- 12578966 TI - Ecological food webs: high-quality data facilitate theoretical unification. PMID- 12578967 TI - Insights into mRNA transport in neurons. PMID- 12578969 TI - Exonucleolytic degradation of double-stranded RNA by an activity in Xenopus laevis germinal vesicles. AB - We have identified, in extracts from Xenopus laevis germinal vesicles, a 5' exonuclease activity that cleaves double-stranded RNA (dsRNA). Features of the 5' ends of dsRNAs determine whether the strands are symmetrically or asymmetrically degraded. The activity hydrolyzes in the 5' to 3' direction, releasing 5' mononucleotides processively, favoring strands with 5'-monophosphate termini; molecules with capped ends are resistant to digestion. Because of its ability to processively digest dsRNA to mononucleotides, we have named the exonuclease Chipper, which could cooperate or compete with Dicer (an endonuclease that produces molecules with a 5'-phosphate) in the processing of dsRNA. PMID- 12578968 TI - Patterns of DNA sequence polymorphism at Sod vicinities in Drosophila melanogaster: unraveling the footprint of a recent selective sweep. AB - We survey DNA sequence polymorphisms at the Sod locus and four neighboring regions of Drosophila melanogaster, spanning 55,513 base pairs (bp), in 15 strains from a natural population, plus one reference laboratory strain and one strain of Drosophila simulans. Our objective is to characterize a proposed selective sweep that occurred at a locus close to Sod in D. melanogaster and to characterize the strength of the selection event, its time, and the size of the hitchhiked region. Two regions, 1819 and 6kbr3r, show a pattern of polymorphism very similar to the one of Sod, implying that they have been affected by the same evolutionary process that impacted Sod. A third fragment, 2021 seems unaffected by the event. A fourth one, 4039, on the opposite flank of Sod in relation to 2021, is only partially affected. We estimate that the length of the chromosomal segment impacted by the selective sweep is 41-54 kb, the age of the selective sweep is 2,600-22,000 years, and the selective advantage is 0.020 < s < 0.103. PMID- 12578970 TI - cAMP-dependent protein kinase phosphorylation of the acid-sensing ion channel-1 regulates its binding to the protein interacting with C-kinase-1. AB - The acid-sensing ion channel-1 (ASIC1) contributes to synaptic plasticity and may influence the response to cerebral ischemia and acidosis. We found that cAMP dependent protein kinase phosphorylated heterologously expressed ASIC1 and endogenous ASIC1 in brain slices. ASIC1 also showed significant phosphorylation under basal conditions. Previous studies showed that the extreme C-terminal residues of ASIC1 bind the PDZ domain of the protein interacting with C-kinase-1 (PICK1). We found that protein kinase A phosphorylation of Ser-479 in the ASIC1 C terminus interfered with PICK1 binding. In contrast, minimizing phosphorylation or mutating Ser-479 to Ala enhanced PICK1 binding. Phosphorylation-dependent disruption of PICK1 binding reduced the cellular colocalization of ASIC1 and PICK1. Thus, the ASIC1 C terminus contains two sites that influence its binding to PICK1. Regulation of this interaction by phosphorylation provides a mechanism to control the cellular localization of ASIC1. PMID- 12578971 TI - Individuality and variation in gene expression patterns in human blood. AB - The nature and extent of interindividual and temporal variation in gene expression patterns in specific cells and tissues is an important and relatively unexplored issue in human biology. We surveyed variation in gene expression patterns in peripheral blood from 75 healthy volunteers by using cDNA microarrays. Characterization of the variation in gene expression in healthy tissue is an essential foundation for the recognition and interpretation of the changes in these patterns associated with infections and other diseases, and peripheral blood was selected because it is a uniquely accessible tissue in which to examine this variation in patients or healthy volunteers in a clinical setting. Specific features of interindividual variation in gene expression patterns in peripheral blood could be traced to variation in the relative proportions of specific blood cell subsets; other features were correlated with gender, age, and the time of day at which the sample was taken. An analysis of multiple sequential samples from the same individuals allowed us to discern donor specific patterns of gene expression. These data help to define human individuality and provide a database with which disease-associated gene expression patterns can be compared. PMID- 12578972 TI - The origin of the boson peak and thermal conductivity plateau in low-temperature glasses. AB - We argue that the intrinsic glassy degrees of freedom in amorphous solids giving rise to the thermal conductivity plateau and the "boson peak" in the heat capacity at moderately low temperatures are directly connected to those motions giving rise to the two-level-like excitations seen at still lower temperatures. These degrees of freedom can be thought of as strongly anharmonic transitions between the local minima of the glassy energy landscape that are accompanied by ripplon-like domain wall motions of the glassy mosaic structure predicted to occur at T(g) by the random first-order transition theory. The energy spectrum of the vibrations of the mosaic depends on the glass transition temperature, the Debye frequency, and the molecular length scale. The resulting spectrum reproduces the experimental low-temperature boson peak. The "nonuniversality" of the thermal conductivity plateau depends on k(B)T(g)omega(D) and arises from calculable interactions with the phonons. PMID- 12578973 TI - Effects of C-terminal deletions on cystic fibrosis transmembrane conductance regulator function in cystic fibrosis airway epithelia. AB - To better understand the function of the conserved C terminus of the cystic fibrosis (CF) transmembrane conductance regulator, we studied constructs containing deletions in the C-terminal tail. When expressed in well differentiated CF airway epithelia, each construct localized predominantly to the apical membrane and generated transepithelial Cl(-) current. The results suggested that neither the C-terminal PSD-95/Discs-large/ZO-1 (PDZ)-interacting motif nor other C-terminal sequences were absolutely required for apical expression in airway epithelia. Surprisingly, deleting an acidic cluster near the C terminus reduced both channel opening rate and transepithelial Cl(-) transport, indicating that it influences channel gating. These results may help explain the relative paucity of CF-associated mutations in the C terminus. PMID- 12578974 TI - The effect of anandamide on prolactin secretion is modulated by estrogen. AB - Recent research has revealed that endogenous cannabinoid receptors (CB1 and CB2) react with the active ingredient of marijuana, Delta(9)-tetrahydrocannabinol. Two endogenous ligands activate these receptors. The principal one, anandamide (AEA), activates CB1. AEA and CB1 are localized to various neurons within the brain. Because Delta(9)-tetrahydrocannabinol inhibited prolactin (Prl) secretion following its intraventricular injection into male rats, we hypothesized that AEA would have a similar effect. Estrogen modifies many hormonal responses and is known to increase Prl secretion. Therefore, we hypothesized that responses to intraventricular AEA would change depending on the gonadal steroid environment. Consequently, we evaluated the effects of lateral cerebral ventricular microinjection of AEA (20 ng) into male, ovariectomized (OVX), and estrogen primed (OVX-E) rats. AEA decreased plasma Prl in male rats, had little effect in OVX females, and increased Prl in OVX-E rats. The results were at least partially mediated by changes in dopaminergic turnover, altering the inhibitory dopaminergic control of Prl release by the anterior pituitary gland. Thus, dopamine turnover was increased in the male rats and decreased significantly in OVX and in OVX-E rats. The changes in Prl may be caused not only by altered dopamine input to the anterior pituitary gland but also by effects of AEA on other transmitters known to alter Prl release. Importantly, in OVX-E rats, the elevated Prl release and the response to AEA were blocked by the AEA antagonist, indicating that AEA is a synaptic transmitter released from neurons that decrease inhibitory control of Prl release. PMID- 12578975 TI - Insights into glutamate transport regulation in human astrocytes: cloning of the promoter for excitatory amino acid transporter 2 (EAAT2). AB - Glutamate transport is central to neurotransmitter functions in the brain. Impaired glutamate transport induces neurotoxicity associated with numerous pathological processes, including stroke/ischemia, temporal lobe epilepsy, Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, HIV-1 associated dementia, and growth of malignant gliomas. Excitatory amino acid transporter-2 (EAAT2) is a major glutamate transporter in the brain expressed primarily in astrocytes. We presently describe the cloning and characterization of the human EAAT2 promoter, demonstrating elevated expression in astrocytes. Regulators of EAAT2 transport, both positive and negative, alter EAAT2 transcription, promoter activity, mRNA, and protein. These findings imply that transcriptional processes can regulate EAAT2 expression. Moreover, they raise the intriguing possibility that the EAAT2 promoter may be useful for targeting gene expression in the brain and for identifying molecules capable of modulating glutamate transport that could potentially inhibit, ameliorate, or prevent various neurodegenerative diseases. PMID- 12578978 TI - Transfer of a passive additive in a turbulent boundary layer at very large Reynolds numbers. AB - We formulate the mass transfer problem for a passive additive in a turbulent boundary layer based on the recently proposed model of the turbulent boundary layer at very large Reynolds numbers. The solutions of three basic problems are obtained. These solutions are self-similar asymptotics describing the mass exchange at its initial stages. The solutions obtained can be used for the construction (in particular, the numerical construction) of the solution to the more general problems of passive admixture transfer in the developed turbulent wall-bounded shear flows. PMID- 12578977 TI - Multiple routes to memory: distinct medial temporal lobe processes build item and source memories. AB - A central function of memory is to permit an organism to distinguish between stimuli that have been previously encountered and those that are novel. Although the medial temporal lobe (which includes the hippocampus and surrounding perirhinal, parahippocampal, and entorhinal cortices) is known to be crucial for recognition memory, controversy remains regarding how the specific subregions within the medial temporal lobe contribute to recognition. We used event-related functional MRI to examine the relation between activation in distinct medial temporal lobe subregions during memory formation and the ability (i) to later recognize an item as previously encountered (item recognition) and (ii) to later recollect specific contextual details about the prior encounter (source recollection). Encoding activation in hippocampus and in posterior parahippocampal cortex predicted later source recollection, but was uncorrelated with item recognition. In contrast, encoding activation in perirhinal cortex predicted later item recognition, but not subsequent source recollection. These outcomes suggest that the subregions within the medial temporal lobe subserve distinct, but complementary, learning mechanisms. PMID- 12578976 TI - Differential effects of prostaglandin derived from omega-6 and omega-3 polyunsaturated fatty acids on COX-2 expression and IL-6 secretion. AB - Omega-6 (omega-6) polyunsaturated fatty acids (PUFA), abundant in the Western diet, are precursors for a number of key mediators of inflammation including the 2-series of prostaglandins (PG). PGE(2), a cyclooxygenase (COX) metabolite of arachidonic acid, a omega-6 PUFA, is a potent mediator of inflammation and cell proliferation. Dietary supplements rich in omega-3 PUFA reduce the concentrations of 2-series PG and increase the synthesis of 3-series PG (e.g., PGE(3)), which are believed to be less inflammatory. However, studies on cellular consequences of increases in 3-series PG in comparison to 2-series PG have not been reported. In this study, we compared the effects of PGE(2) and PGE(3) on (i) cell proliferation in NIH 3T3 fibroblasts, (ii) expression and transcriptional regulation of the COX-2 gene in NIH 3T3 fibroblasts, and (iii) the production of an inflammatory cytokine, IL-6, in RAW 264.7 macrophages. PGE(3), unlike PGE(2), is not mitogenic to NIH 3T3 fibroblasts. PGE(2) and PGE(3) both induce COX-2 mRNA via similar signaling mechanisms; however, compared with PGE(2), PGE(3) is significantly less efficient in inducing COX-2 gene expression. Furthermore, although both PGE(2) and PGE(3) induce IL-6 synthesis in RAW 264.7 macrophages, PGE(3) is substantially less efficient compared with PGE(2). We further show that increasing the omega-3 content of membrane phospholipid results in a decrease in mitogen-induced PGE(2) synthesis. Taken together, our data suggest that successful replacement of omega-6 PUFA with omega-3 PUFA in cell membranes can result in a decreased cellular response to mitogenic and inflammatory stimuli. PMID- 12578979 TI - Regulation of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1 alpha ) and mitochondrial function by MEF2 and HDAC5. AB - The myocyte enhancer factor-2 (MEF2) transcription factor regulates muscle development and calcium-dependent gene expression. MEF2 activity is repressed by class II histone deacetylases (HDACs), which dissociate from MEF2 when phosphorylated on two serine residues in response to calcium signaling. To explore the potential importance of MEF2/HDAC interactions in the heart, we generated transgenic mice expressing a signal-resistant form of HDAC5 under cardiac-specific and doxycycline-inducible regulation. Transgene expression resulted in sudden death in male mice accompanied by loss and morphologic changes of cardiac mitochondria and down-regulation of mitochondrial enzymes. The transcriptional coactivator PGC-1 alpha, a master regulator of mitochondrial biogenesis and fatty acid oxidation, was also down-regulated in response to HDAC5 expression. Examination of the PGC-1 alpha promoter revealed two MEF2-binding sites that mediate transcriptional activation by MEF2 and repression by HDAC5. These findings identify PGC-1 alpha as a key target of the MEF2/HDAC regulatory pathway and demonstrate this pathway's importance in maintenance of cardiac mitochondrial function. PMID- 12578980 TI - Multiparameter single-molecule fluorescence spectroscopy reveals heterogeneity of HIV-1 reverse transcriptase:primer/template complexes. AB - By using single-molecule multiparameter fluorescence detection, fluorescence resonance energy transfer experiments, and newly developed data analysis methods, this study demonstrates directly the existence of three structurally distinct forms of reverse transcriptase (RT):nucleic acid complexes in solution. Single molecule multiparameter fluorescence detection also provides first information on the structure of a complex not observed by x-ray crystallography. This species did not incorporate nucleotides and is structurally distinct from the other two observed species. We determined that the nucleic acid substrate is bound at a site far removed from the nucleic acid-binding tract observed by crystallography. In contrast, the other two states are identified as being similar to the x-ray crystal structure and represent distinct enzymatically productive stages in DNA polymerization. These species differ by only a 5-A shift in the position of the nucleic acid. Addition of nucleoside triphosphate or of inorganic pyrophosphate allowed us to assign them as the educt and product state in the polymerization reaction cycle; i.e., the educt state is a complex in which the nucleic acid is positioned to allow nucleotide incorporation. The second RT:nucleic acid complex is the product state, which is formed immediately after nucleotide incorporation, but before RT translates to the next nucleotide. PMID- 12578981 TI - The Escherichia coli multidrug transporter MdfA catalyzes both electrogenic and electroneutral transport reactions. AB - The resistance of cells to many drugs simultaneously (multidrug resistance) often involves the expression of membrane transporters (Mdrs); each recognizes and expels a broad spectrum of chemically unrelated drugs from the cell. The Escherichia coli Mdr transporter MdfA is able to transport differentially charged substrates in exchange for protons. This includes neutral compounds, namely chloramphenicol and thiamphenicol, and lipophilic cations such as tetraphenylphosphonium and ethidium. Here we show that the chloramphenicol and thiamphenicol transport reactions are electrogenic, whereas the transport of several monovalent cationic substrates is electroneutral. Therefore, unlike with positively charged substrates, the transmembrane electrical potential (negative inside) constitutes a major part of the driving force for the transport of electroneutral substrates by MdfA. These results demonstrate an unprecedented ability of a single secondary transporter to catalyze discrete transport reactions that differ in their electrogenicity and are governed by different components of the proton motive force. PMID- 12578982 TI - Visualization of synaptotagmin I oligomers assembled onto lipid monolayers. AB - Neuronal exocytosis is mediated by Ca(2+)-triggered rearrangements between proteins and lipids that result in the opening and dilation of fusion pores. Synaptotagmin I (syt I) is a Ca(2+)-sensing protein proposed to regulate fusion pore dynamics via Ca(2+)-promoted binding of its cytoplasmic domain (C2A-C2B) to effector molecules, including anionic phospholipids and other copies of syt. Functional studies indicate that Ca(2+)-triggered oligomerization of syt is a critical step in excitation-secretion coupling; however, this activity has recently been called into question. Here, we show that Ca(2+) does not drive the oligomerization of C2A-C2B in solution. However, analysis of Ca(2+).C2A-C2B bound to lipid monolayers, using electron microscopy, revealed the formation of ring like heptameric oligomers that are approximately 11 nm long and approximately 11 nm in diameter. In some cases, C2A-C2B also assembled into long filaments. Oligomerization, but not membrane binding, was disrupted by neutralization of two lysine residues (K326,327) within the C2B domain of syt. These data indicate that Ca(2+) first drives C2A-C2B.membrane interactions, resulting in conformational changes that trigger a subsequent C2B-mediated oligomerization step. Ca(2+) mediated rearrangements between syt subunits may regulate the opening or dilation kinetics of fusion pores or may play a role in endocytosis after fusion. PMID- 12578983 TI - Base pairing among three cis-acting sequences contributes to template switching during hepadnavirus reverse transcription. AB - Synthesis of the relaxed-circular (RC) DNA genome of hepadnaviruses requires two template switches during plus-strand DNA synthesis: primer translocation and circularization. Although primer translocation and circularization use different donor and acceptor sequences, and are distinct temporally, they share the common theme of switching from one end of the minus-strand template to the other end. Studies of duck hepatitis B virus have indicated that, in addition to the donor and acceptor sequences, three other cis-acting sequences, named 3E, M, and 5E, are required for the synthesis of RC DNA by contributing to primer translocation and circularization. The mechanism by which 3E, M, and 5E act was not known. We present evidence that these sequences function by base pairing with each other within the minus-strand template. 3E base-pairs with one portion of M (M3) and 5E base-pairs with an adjacent portion of M (M5). We found that disrupting base pairing between 3E and M3 and between 5E and M5 inhibited primer translocation and circularization. More importantly, restoring base pairing with mutant sequences restored the production of RC DNA. These results are consistent with the model that, within duck hepatitis B virus capsids, the ends of the minus strand template are juxtaposed via base pairing to facilitate the two template switches during plus-strand DNA synthesis. PMID- 12578984 TI - Regulatory gene networks and the properties of the developmental process. AB - Genomic instructions for development are encoded in arrays of regulatory DNA. These specify large networks of interactions among genes producing transcription factors and signaling components. The architecture of such networks both explains and predicts developmental phenomenology. Although network analysis is yet in its early stages, some fundamental commonalities are already emerging. Two such are the use of multigenic feedback loops to ensure the progressivity of developmental regulatory states and the prevalence of repressive regulatory interactions in spatial control processes. Gene regulatory networks make it possible to explain the process of development in causal terms and eventually will enable the redesign of developmental regulatory circuitry to achieve different outcomes. PMID- 12578985 TI - Regulation of photoperiodic flowering by Arabidopsis photoreceptors. AB - Photoperiodism is a day-length-dependent seasonal change of physiological or developmental activities that is widely found in plants and animals. Photoperiodic flowering in plants is regulated by photosensory receptors including the red/far-red light-receptor phytochromes and the blue/UV-A light receptor cryptochromes. However, the molecular mechanisms underlying the specific roles of individual photoreceptors have remained poorly understood. Here, we report a study of the day-length-dependent response of cryptochrome 2 (cry2) and phytochrome A (phyA) and their role as day-length sensors in Arabidopsis. The protein abundance of cry2 and phyA showed a diurnal rhythm in plants grown in short-day but not in plants grown in long-day. The short-day-specific diurnal rhythm of cry2 is determined primarily by blue light-dependent cry2 turnover. Consistent with a proposition that cry2 and phyA are the major day-length sensors in Arabidopsis, we show that phyA mediates far-red light promotion of flowering with modes of action similar to that of cry2. Based on these results and a finding that the photoperiodic responsiveness of plants depends on light quality, a model is proposed to explain how individual phytochromes and cryptochromes work together to confer photoperiodic responsiveness in Arabidopsis. PMID- 12578986 TI - MyoD is functionally linked to the silencing of a muscle-specific regulatory gene prior to skeletal myogenesis. AB - Most of the genes that are central to the process of skeletal muscle differentiation remain in a transcriptionally silent or "off" state until muscle cells (myoblasts) are induced to differentiate. Although the mechanisms that contribute to this phenomenon are still unclear, it is likely that histone deacetylases (HDACs), which play an important role in the repression of genes, are principally involved. Recent studies indicate that the initiator of the myogenic program, namely MyoD, can associate with the deacetylase HDAC1 in vivo, and because HDACs are usually recruited to promoters by specific proteins, we considered the possibility that these two proteins might be acting together at the promoters of muscle-specific genes to repress their transcription in myoblasts. In this work, we show by chromatin immunoprecipitation (ChIP) assays that MyoD and HDAC1 are both occupying the promoter of myogenin and that this gene is in a region of repressed chromatin, as revealed by enrichment in histone H3 lysine 9 (Lys-9) methylation and the underacetylation of histones. Surprisingly, after the myoblasts are induced to differentiate, the promoter becomes absent of HDAC1, and eventually the acetyltransferase P/CAF takes it place alongside MyoD. In addition, enrichment of histone H3 acetylation (Lys 9/14) and phosphorylation of Ser-10 can now be observed at the myogenin promoter. These data strongly suggest that in addition to its widely accepted role as an activator of differentiation-specific genes, MyoD also can perform as a transcriptional repressor in proliferating myoblasts while in partnership with a HDAC. PMID- 12578987 TI - Molecular bases of defective signal transduction in the platelet P2Y12 receptor of a patient with congenital bleeding. AB - We have identified structural attributes required for signal transduction through a seven-transmembrane-domain receptor. Platelets from a patient (AC) with a congenital bleeding disorder had normal shape change but reduced and reversible aggregation in response to 4 microM ADP, similar to normal platelets with blocked P2Y(12) receptor. The response to 20 microM ADP, albeit still decreased, was more pronounced and was reduced by a P2Y(12) antagonist, indicating some residual receptor function. ADP failed to lower the adenylyl cyclase activity stimulated by prostaglandin E(1) in the patient's platelets, even though the number and affinity of 2-methylthioadenosine 5'-[(33)P]diphosphate-binding sites was normal. Analysis of the patient's P2Y(12) gene revealed a G-to-A transition in one allele, changing the codon for Arg-256 in the sixth transmembrane domain to Gln, and a C-to-T transition in the other allele, changing the codon for Arg-265 in the third extracellular loop to Trp. Neither mutation interfered with receptor surface expression but both altered function, since ADP inhibited the forskolin induced increase of cAMP markedly less in cells transfected with either mutant P2Y(12) as compared with wild-type receptor. These studies delineate a region of P2Y(12) required for normal function after ADP binding. PMID- 12578988 TI - Hypersusceptibility of cystic fibrosis mice to chronic Pseudomonas aeruginosa oropharyngeal colonization and lung infection. AB - No transgenic cystic fibrosis (CF) mouse model developed to date mimics the major clinical phenotype found in humans with CF, chronic Pseudomonas aeruginosa lung infection. In a transgenic CF transmembrane conductance regulator (cftr) mouse colony, we found WT, heterozygous, and homozygous CF mice housed in the same cage became chronically colonized in the oropharynx with environmental P. aeruginosa when the bacterium was present in drinking water. Elimination of P. aeruginosa from drinking water resulted in clearance in most WT and CF heterozygous, but not homozygous mice. For experimental evaluation, a combination of specific animal husbandry techniques and an oral infection route showed cftr(-/-) mice but not WT mice can be chronically colonized by P. aeruginosa with subsequent lung translocation, yielding a pathologic picture indicative of chronic lung infection. In some instances, mucoid isolates of P. aeruginosa were recovered from lungs, indicating conditions were present for conversion to mucoidy. Overexpression of human CFTR in the lungs of WT mice markedly accelerated the clearance rate of P. aeruginosa, demonstrating that lung levels of CFTR play an important role in defense against infection. P. aeruginosa mutants unable to express the surface polysaccharide alginate or the global regulator GacA were deficient in their ability to colonize the mice. CF mice made potent immune responses to P. aeruginosa outer membrane antigens. Overall, we found that under the proper conditions, transgenic CF mice are hypersusceptible to P. aeruginosa colonization and infection and can be used for evaluations of lung pathophysiology, bacterial virulence, and development of therapies aimed at treating CF lung disease. PMID- 12578989 TI - Arabidopsis LEAFY COTYLEDON1 represents a functionally specialized subunit of the CCAAT binding transcription factor. AB - LEAFY COTYLEDON1 (LEC1) is a central regulator that is required for many aspects of Arabidopsis embryogenesis and sufficient to induce embryo development in vegetative cells when expressed ectopically. We previously showed that LEC1 encodes an HAP3 subunit of the CCAAT binding factor and that the 10 Arabidopsis HAP3 (AHAP3) subunits can be divided into two classes, the LEC1-type and the non LEC1-type, based on sequence similarity within their B domains. By analyzing the ability of chimeric HAP3 subunits containing different combinations of domains from LEC1 and a non-LEC1-type AHAP3 subunit to suppress genetically the lec1 mutation, we show that the B domain of LEC1 is necessary and sufficient within the context of the protein for its activity in embryogenesis. Moreover, we identify one amino acid residue, Asp-55, specific to the LEC1-type B domain that is required for LEC1 activity in embryogenesis and sufficient to confer partial LEC1 activity to a non-LEC1-type B domain. Based on structural similarities between the HAP3 B domain and histone fold motif, we discuss how the Asp-55 residue may functionally differentiate LEC1 from the non-LEC1-type AHAP3 subunits. PMID- 12578990 TI - Electrophile and oxidant damage of mitochondrial DNA leading to rapid evolution of homoplasmic mutations. AB - mtDNA mutations occur in a wide variety of degenerative diseases and cancer. mtDNA seems to be more susceptible to DNA damage and consequently sustains higher rates of mutation than does nuclear DNA (nDNA). Many of the somatic mtDNA mutations in human cancers are located in the displacement loop (D-loop) and in particular in a polycytidine stretch (C-tract) termed D310. The D310 region exhibits polymorphic length variation among individuals and has been described as a "hot spot" for somatic mutations in many cancer types. We used real-time quantitative PCR to analyze mtDNA integrity, damage repair, and induced mutations after exposure of human adult retinal pigment epithelial (ARPE)-19 cells to 4 nitroquinoline 1-oxide, a UV-mimetic and adduct-forming carcinogen, and tert butyl hydroperoxide, an oxidant. The mtDNA-damage profile depended on the region. Thus, the tRNA coding for glycine (tRNA(G)) was the least affected region, whereas the D-loop, and especially its D310 region, were most sensitive to damage. The time course of repair of mutations of the D-loop and especially the D310 region after exposure to DNA-damaging agents was delayed when compared with other regions and gave rise to common D310 C-tract frame-shift mutations. The induced mutations in the D310 region were predominantly homoplasmic only 7 days after exposure to damage. Our results establish that the D-loop (especially its D310 region) is highly susceptible to mutations because of its vulnerability to DNA damage and inefficient repair mechanisms. Our findings may explain the high frequency of homoplasmic D310 somatic mutations in many tumor types. PMID- 12578991 TI - The structural basis of cysteine aminoacylation of tRNAPro by prolyl-tRNA synthetases. AB - Cysteinyl-tRNA synthetase is an essential enzyme required for protein synthesis. Genes encoding this protein have not been identified in Methanocaldococcus jannaschii, Methanothermobacter thermautotrophicus, or Methanopyrus kandleri. It has previously been proposed that the prolyl-tRNA synthetase (ProRS) enzymes in these organisms recognize either proline or cysteine and can aminoacylate their cognate tRNAs through a dual-specificity mechanism. We report five crystal structures at resolutions between 2.6 and 3.2 A: apo M. jannaschii ProRS, and M. thermautotrophicus ProRS in apo form and in complex with cysteinyl-sulfamoyl-, prolyl-sulfamoyl-, and alanyl-sulfamoyl-adenylates. These aminoacyl-adenylate analogues bind to a single active-site pocket and induce an identical set of conformational changes in loops around the active site when compared with the ligand-free conformation of ProRS. The cysteinyl- and prolyl-adenylate analogues have similar, nanomolar affinities for M. thermautotrophicus ProRS. Homology modeling of tRNA onto these adenylate complexes places the 3'-OH of A76 in an appropriate position for the transfer of any of the three amino acids to tRNA. Thus, these structures explain recent biochemical experiments showing that M. jannaschii ProRS misacylates tRNA(Pro) with cysteine, and argue against the proposal that these archaeal ProRS enzymes possess the dual capacity to aminoacylate both tRNA(Pro) and tRNA(Cys) with their cognate amino acids. PMID- 12578992 TI - Introduction for Dr. Richard A. Brand, Jr. PMID- 12578994 TI - Bucket-handle tear of acetabular labrum accompanying posterior dislocation of the hip. From J Bone Joint Surg 41A:131-134, 1959. PMID- 12578995 TI - Assessment and differential diagnosis of the painful hip. AB - Hip pain is a common problem seen by orthopaedic surgeons. The current authors provide an approach to the patient with hip pain, including important information to be gained from the history and physical examination and relevant radiographic studies and laboratory tests. A differential diagnosis for patients presenting with the complaint of hip pain and indications for hip arthroscopy are provided. PMID- 12578996 TI - Imaging the painful hip. AB - With the advent of magnetic resonance imaging and, subsequently, magnetic resonance arthrography, the imaging algorithm for hip pain has evolved considerably. Magnetic resonance imaging has supplanted bone scintigraphy as the first line imaging test after conventional radiographs in the setting of suspected occult fracture, transient marrow edema, and osteonecrosis. Computed tomography scanning and magnetic resonance imaging are invaluable for the evaluation of monarticular arthropathies such as pigmented villonodular synovitis and synovial osteochondromatosis. By combining conventional magnetic resonance imaging with capsular distention afforded by arthrography, magnetic resonance arthrography has become the imaging examination of choice for disorders of the acetabular labrum and for the evaluation of articular cartilage at the hip. PMID- 12578997 TI - Hip arthroscopy: surgical approach, positioning, and distraction. AB - The hip is the most deeply recessed joint in the body making it difficult to access arthroscopically. However, for the patient with a symptomatic loose body or labral tear open techniques are less advantageous. It is for this reason that, however difficult, minimally-invasive approaches to the hip have been developed. Meticulous attention to proper positioning and portal placement is paramount for safe and successful arthroscopy of the hip. The current authors will review the seminal features of positioning and minimally-invasive surgical approaches to the hip. PMID- 12578998 TI - Anatomy, pathologic features, and treatment of acetabular labral tears. AB - There is an evolving body of knowledge regarding the acetabular labrum. Labral tears are most frequently anterior and often are associated with sudden twisting or pivoting motions. High clinical suspicion in association with positive physical findings are fundamental for the clinician to properly determine treatment for the suspected tear. Labral tears, especially those present for years, may contribute to the progression of hip osteoarthritis. Patients at risk include those with developmental dysplasia, those with tears greater than 5 years, and those with associated chondral full-thickness lesions. Chondral injuries may occur in association with a multitude of hip conditions including labral tears, loose bodies, osteonecrosis, slipped capital femoral epiphysis, dysplasia, and degenerative arthritis. Labral tears occurring at the watershed zone may destabilize the adjacent acetabular conditions. Arthroscopic observations support the concept that labral disruption, acetabular chondral lesions, or both frequently are part of a continuum of degenerative joint disease. PMID- 12578999 TI - The role of hip arthroscopy in the treatment of synovial disorders and loose bodies. AB - Arthroscopic surgery of the hip is an evolving procedure that can facilitate the diagnosis and treatment of synovial-based disease. Radiographic studies have not reproducibly identified many of the synovial conditions that can affect the hip, and arthroscopy has provided a means to improved diagnostic accuracy and the ability to treat certain conditions. Arthroscopic intervention has been reported in synovial chondromatosis and osteochondromatosis, pigmented villonodular synovitis, inflammatory arthropathies including rheumatoid arthritis, and acute septic arthritis. Patients with other conditions that result in acute and chronic synovitis within the hip such as hemosiderotic synovitis secondary to hemophilia and chondrocalcinosis also may benefit from arthroscopic intervention. These synovial and intraarticular abnormalities have been associated with accelerated joint degeneration, and although the pathologic mechanisms have not been delineated clearly, the clinical description has been advanced using hip arthroscopy. Increased awareness of the association between synovial abnormalities and degeneration has resulted from arthroscopy applied to the hip in early stages of disease and early in the course of symptomatic dysfunction. The indications and treatment capabilities will continue to expand with additional understanding of early hip disease, improvements in equipment, and as outcome studies reveal that the patient benefits from the procedure. PMID- 12579000 TI - Hip arthroscopy in the pediatric population. AB - Hip arthroscopy has obvious advantages over arthrotomy in the pediatric population. Hip arthroscopy, used as a diagnostic or therapeutic tool, is significantly less invasive than arthrotomy and allows for quicker recovery and return to activities. In addition, arthroscopy avoids dislocation of the femoral head and the corresponding risk of osteonecrosis. Current indications for hip arthroscopy in pediatric patients include septic arthritis, labral disorders, slipped capital femoral epiphysis, and Legg-Calve-Perthes disease. The role of hip arthroscopy in the pediatric population will continue to expand because of its attractiveness as a less invasive option for evaluating the hip. PMID- 12579001 TI - Articular cartilage changes in avascular necrosis: an arthroscopic evaluation. AB - Treatment methods for osteonecrosis (avascular necrosis) are wide and varied. When untreated, progression of the disease is common and may dictate femoral head replacement. However, before femoral head collapse, some patients have mechanical joint symptoms (locking, buckling, clicking) that are unaddressed by femoral head drilling alone. Radiographic examinations in these patients usually are nondiagnostic. Patients with these clinical criteria were evaluated arthroscopically. Between 1993 and 2000, seven patients were identified with known documented or radiographic diagnosis of avascular necrosis who had hip arthroscopy. Each patient's preoperative history, physical examination, plain radiographs, magnetic resonance imaging scans, and operative notes were reviewed from a prospectively-derived database. The duration and onset of symptoms were identified carefully. Case histories are presented on five of these patients. Articular cartilage changes were recorded and correlated to the preoperative radiologic studies. Hip arthroscopy is a minimally invasive, highly effective, joint-preserving surgery in the young patient with mechanical symptoms (locking, catching, buckling) and early avascular necrosis. Treatable lesions include loose bodies, synovitis, chondral flaps, and labral tears. In addition, accurate staging can be accomplished through direct observation. PMID- 12579002 TI - The role of hip arthroscopy in the elite athlete. AB - Intraarticular hip disorders in the elite athlete are a relatively rare but serious potential consequence of high-level competition. Axial and torsional forces involving the hips of elite athletes place them at potentially higher risk for chondral injuries, labral injuries, or both. Ten patients (13 hips) had arthroscopy. Of the 10 patients, the average age was 24 years. Nine patients were men and one was a woman. Of the 13 (10 patients) arthroscopies done two cases were bilateral, and one patient had the same hip operated on twice. Seven of the patients were professional hockey players, one patient was a football player, one patient was a baseball player, and one patient was a golfer. All 13 hips (10 patients) had anterior labral tears, whereas two hips had anterior and posterior labral tears. Two hips had an average of four loose bodies, four had evidence of chondral lesions, and one had an anterior margin acetabular fracture. Twelve of 13 arthroscopies were successful; however, one patient had recurrent symptoms. There were no surgical complications. Hip arthroscopy is a safe and reproducible method to diagnose and treat intraarticular hip disorders in athletes, which facilitates earlier return to their respective sport. PMID- 12579003 TI - The nonarthritic hip score: reliable and validated. AB - The purpose of the current study was to assess the validity, internal consistency, and reproducibility of a short, self-administered hip score designed for use in younger patients with higher demands and expectations than older patients with degenerative joint disease. Validity and internal consistency was studied with 48 consecutive patients with a mean age of 33 years with intractable hip pain and normal plain radiographs. Reproducibility was assessed from data on an additional random sample of 17 patients with hip pain. The Pearson correlation coefficients were 0.82 and 0.59 between the nonarthritic hip score and the Harris hip score and Short Form-12, respectively showing validity. Cronbach's coefficient alpha measuring the internal consistency within each of the score's four domains ranged from 0.69 to 0.92. The test and retest reproducibility ranged from 0.87 to 0.95 for the four subsets and was 0.96 overall. This short, self administered questionnaire regarding hip pain in young patients with increased activity demands and high treatment expectations is valid compared with previous measures of hip performance, is internally consistent, and is reproducible. PMID- 12579004 TI - Hip arthroscopy: complications in 1054 cases. AB - Although arthroscopic surgery of the hip is becoming a more frequently done operation, there is relatively few data in the literature regarding the frequency of complications. In the current prospective study of 1054 consecutive hip arthroscopies, the authors report an overall complication rate of 1.4%. Complications included neurapraxia, portal wound bleeding, portal hematoma, trochanteric bursitis, and instrument breakage. The only major complication was one case of septic arthritis. It is concluded that, done properly, hip arthroscopy can be a considered a low-risk procedure. PMID- 12579005 TI - Effect of arthroscopic procedures on the acromioclavicular joint. AB - The objective of the current study was to determine the effect of arthroscopic acromioplasty, and combined acromioplasty and distal clavicle resection on joint kinematics and in situ forces in response to an anterior, posterior, and superior load of 70 N. The loading conditions were applied to 10 fresh-frozen cadaveric shoulders using a robotic and universal force and moment sensor testing system. Translations in response to a posterior load increased by approximately 30% after combined acromioplasty and distal clavicle resection when compared with the intact and acromioplasty conditions. The in situ force in the trapezoid and conoid ligaments increased significantly from 13 +/- 15 N to 40 +/- 25 N and 13 +/- 13 N to 38 +/- 28 N, respectively, between the intact and combined acromioplasty and distal clavicle resection conditions during anterior loading. The results suggest that an arthroscopic acromioplasty alone does not significantly affect the mechanics of the acromioclavicular joint with these loading conditions. However, an acromioplasty combined with a distal clavicular resection does result in significant increases in joint motion and ligament forces. In some circumstances, such as after a previous joint separation, the increased forces in the coracoclavicular ligaments could result in additional damage to weak ligaments. PMID- 12579006 TI - Spontaneous divergent elbow dislocation after Sauve-Kapandji procedure. AB - This is a report on an unusual complication of the Sauve-Kapandji procedure in patients with rheumatoid arthritis. Two women with rheumatoid arthritis who previously had an ipsilateral Sauve-Kapandji procedure experienced spontaneous transverse divergent elbow dislocations without evident trauma. Their radiographs showed medial dislocation of the proximal ulna, which was separated from the radial head. The radial head and distal end of the ulnar shaft showed remarkable instability by a pronation and supination motion without the radial and ulnar shafts being separated from each other. Stress radiographic examination showed significant loosening of all ligaments except the medial collateral ligament around the elbow and did not show disruption of the interosseous membrane. A unique chronic twist radioulnar dissociation which consists of gross instability of the radial head and the distal ulna without disruption of the interosseous membrane was considered to cause instability of the humeroulnar joint, which results in medial dislocation of the proximal ulna. This report suggests that there is a direct cause and effect relationship between the residual distal ulnar instability and the development of transverse divergent dislocation of the elbow in patients with rheumatoid arthritis after the Sauve-Kapandji procedure. PMID- 12579007 TI - Idiopathic avascular necrosis of the metacarpal head. AB - Avascular necrosis of the metacarpal head is a rare condition. Although the condition is associated with trauma, systemic lupus erythematosus, and steroid use, it can occur spontaneously without any obvious cause. Any metacarpal may be affected and the pathologic changes are similar to those described in other bones such as the femur. The diagnosis requires an index of suspicion in a young patient with spontaneous onset of symptoms localized to a metacarpophalangeal joint. Magnetic resonance imaging is a useful early diagnostic tool when the radiographic findings are nonspecific or absent. The natural history of the condition is not known. Although symptoms may resolve with nonoperative treatment, progressive collapse of the metacarpal head and subsequent degenerative arthritis is a possible long-term outcome. Curettage of the lesion and supplementary cancellous bone grafting has been reported to provide symptomatic relief in cases resistant to nonoperative treatment. A case is presented of idiopathic avascular necrosis of the head of the dominant ring finger metacarpal in a 27-year-old woman. The purpose of this report is to highlight the clinical presentation, radiographic features, pathologic findings, and outcome at 2 years after curettage and bone grafting. PMID- 12579008 TI - Familial primary acetabular dysplasia and dislocation of the hip. AB - Radiographic and clinical evaluation was done on 12 members of a kindred that included members with hip dislocation. Acetabular dysplasia was found in several members who did not have dislocation. This supports the concept that acetabular dysplasia is inherited and acts as a precursor to dislocation. Identification of infants with primary acetabular dysplasia may allow measures to prevent dislocation. PMID- 12579009 TI - A study of Korean femoral geometry. AB - The current authors investigated the anatomic geometry of femurs from Korean subjects and compared the results with those of femurs from American and Japanese subjects. Thirty-eight femurs extracted from cadavers and 200 femurs of healthy subjects were used. The 38 femurs were placed on a horizontal plane to obtain computed tomography images, which then were transformed into bitmap images, and the computed tomography images of the 200 femurs of healthy subjects also were obtained. Three-dimensional images of the femurs were reconstructed and used for measuring the parameters. The measurement results suggested that femurs from Korean subjects were different from femurs from American subjects. The major differences were that femurs from Korean subjects had a larger anteversion angle by approximately 8 degrees, a relatively larger canal flare index, and there was a shorter distance between the lesser trochanter and the isthmus. These differences can provide useful information to the design of a new hip prosthesis for Korean, Japanese, and other Asian patients. PMID- 12579010 TI - Can osteonecrosis of the femoral head be recurrent? AB - Three patients with osteonecrosis of the femoral head in whom the extent of necrosis increased by repeated episodes of osteonecrosis are described. In all cases the evidence of two attacks of osteonecrosis was detected clearly on magnetic resonance images. The significance of the recurrent attacks of osteonecrosis is not clear and the detectable incidence of the recurrence seems to be low. The authors propose the possibility that the osteonecrosis may be the result of multiple episodes of necrosis. PMID- 12579011 TI - Magnetic resonance imaging of total knee arthroplasty. AB - Evaluation of painful total knee arthroplasty can be clinically difficult, and traditional imaging techniques such as conventional radiographs, arthrography, and bone scintigraphy are limited by poor contrast resolution and specificity. Traditional magnetic resonance imaging techniques often are nondiagnostic because of significant metal artifact. Forty-one patients (46 knees) had magnetic resonance imaging, tailored to reduce metallic susceptibility artifact, after total knee arthroplasty, and the findings and clinical and surgical followup were reviewed. All studies consistently showed the integrity of the periprosthetic soft tissues. Magnetic resonance imaging findings led to surgical or other therapeutic interventional procedures in 20 patients, and influenced clinical treatment in all patients. Optimized magnetic resonance imaging, in which the metallic artifact is diminished, is a clinically useful adjunct to traditional imaging techniques in evaluation of patients with painful total knee arthroplasty. PMID- 12579012 TI - The proximal tibiofibular joint: an anatomic study. AB - In the first stage of the current study, 14 fresh specimens from above-the-knee amputations were examined by magnetic resonance imaging arthrography. In the second part of the study, these amputation specimens and 38 embalmed knees from cadavers were dissected. The types of the joint and insertion of the biceps femoris tendon and the other structures were observed. Of 14 fresh specimens from above-the-knee amputations examined by magnetic resonance imaging arthrography, nine had a clear communication between the proximal tibiofibular joint and the knee. It was difficult to distinguish the anterior tibiofibular ligament in 30 specimens as a separate band because it was fused intimately with the biceps femoris tendon. Of these specimens, 24 had an oblique type of joint. Because of this communication, the proximal tibiofibular joint might be construed as the fourth compartment of the knee to explain subtle knee problems. PMID- 12579013 TI - Bone resection and ligament treatment for flexion contracture in knee arthroplasty. AB - A retrospective study of 103 knees (88 patients) who had primary total knee arthroplasty with a flexion contracture ranging from 20 degrees to 60 degrees was done to tabulate the primary soft tissue structures released during surgery and to identify any residual deformity. The average flexion contracture preoperatively was 27.1 degrees +/- 8 degrees and postoperatively was 2.7 degrees +/- 3.4 degrees (range, 0 degrees -10 degrees ). The average followup was 70.4 months (range, 12-180 months). Only medial or lateral soft tissue balancing procedures were necessary to correct the flexion contracture in 37 knees (35.9%) and no medial or lateral release was necessary in 25 knees (24.3%), of which 16 had a balanced posterior cruciate ligament. The posterior capsule was released on the deformity side of the knee in 15 knees (14.6%) and on the opposite side of the deformity in seven knees (6.8%). The posterior cruciate ligament was balanced in 21 knees (20.4%) and was released in four knees (3.9%). For all knees in which the posterior cruciate ligament was released or balanced, it was done for excessive rollback and tightness in flexion and not for flexion contracture management. In two patients (2%) an additional 4 mm of distal femur was resected for a 45 degrees and a 25 degrees flexion contracture. The data suggest that a contracted collateral ligament is the most likely primary structure whose effective release allows correction of the flexion contracture in most cases. PMID- 12579014 TI - Proprioception with bicondylar sledge prostheses retaining cruciate ligaments. AB - The current study was designed to evaluate the proprioceptive performance of the knee after implantation of bicondylar sledge prostheses in comparison with contralateral knees and knees of healthy control subjects. After implantation of a sledge prosthesis in the medial and lateral condyles of a knee retaining all ligaments, 15 patients were evaluated clinically and by proprioceptive testing. Clinical examination was done using common scoring systems. Proprioceptive performance was examined using sway measurements during single-leg stance on a force platform. Eleven healthy subjects of the same age served as a control group. Clinical results of both groups differed significantly in all applied scores, except for the subcategories gait, muscle force, and pain. Sway measurements did not differ between right and left legs of subjects in the control group. The patients' surgically treated legs had slightly higher values than the control group, but a statistical significance was not found. Statistical analysis revealed no significant correlation between clinical scores and sway measurement results. Total knee arthroplasty that retains all intraarticular ligaments achieves proprioceptive results comparable with healthy subjects of the same age. PMID- 12579015 TI - Ankle fusion attributable to posttraumatic arthrosis: a long-term followup of 48 patients. AB - The subjective, clinical, functional, and radiologic long-term results of 48 patients after tibiotalar arthrodesis done for posttraumatic and isolated arthrosis of the ankle are reported. After an average followup of 9.3 years, good and very good results were achieved in 92% for subjective parameters and in 73% for clinical and functional parameters as reflected in the American Orthopaedic Foot and Ankle Society ankle and hindfoot score. Fusion of the ankle in a position greater than 5 degrees plantar flexion was correlated with a worse late clinical outcome monitored by the American Orthopaedic Foot and Ankle Society ankle and hindfoot score. At the followup, average tarsal mobility in the surgically treated foot was reduced to 54% of the contralateral side and that loss of tarsal mobility led to a poorer clinical outcome. However, a high incidence of subsequent arthrosis in the adjacent joints of the foot seems not to be completely avoidable in the long-term. The subtalar joint had moderate and severe arthrosis in 47% of patients. Long-term outcome was worse in these patients compared with patients with mild or without degenerative changes in this joint. The current study justifies the value of ankle fusion as a surgical treatment option in patients with end-stage arthrosis in the ankle, provided precise intraoperative positioning of the arthrodesis and the importance of the subtalar joint, are given due consideration. PMID- 12579016 TI - Bone tumors of the extremities or pelvis treated by microwave-induced hyperthermia. AB - From July 1992 through March 1999, 213 patients with malignant bone tumors of the extremities (176 patients) and pelvis (37 patients) were treated by microwave induced hyperthermia. Osteosarcoma and chondrosarcoma were the most common diagnoses. The limb-salvage procedure was done as follows: After separating the tumor-bearing bone and the extraosseous mass from surrounding normal tissues with a proper margin, microwave energy was delivered into the tumor while the healthy tissues were protected carefully from overheating. Restrengthening of the devitalized bone was necessary in many cases. The eschar resulting from the heat necrosis was curettaged. Most patients can walk early with a partial weightbearing brace for support. The survival rate was 73.9%. Fracture, local recurrence, and infection were the main complications although the majority of complications occurred early in the study. Thermotherapy is a novel and effective way to treat bone tumors in selected patients. PMID- 12579017 TI - Slotted intramedullary hip screw nails reduce proximal mechanical unloading. AB - The current prospective and randomized study was done to determine whether fixation of intertrochanteric femoral fractures that lack a medial buttress with a dynamically locked intramedullary hip screw prevents proximal stress shielding. Stress shielding was evident by the presence of cortical hypertrophy at the level of the tip of the nail and often was associated with midthigh pain. Of the 80 patients enrolled in this trial, 64 still were alive after 1 year. Among these patients, 30 had the nail classically locked with two screws transfixing the nail in two separate holes (Group A), and 34 had the nail locked with one screw passing through a slot (Group B). The average duration of followup was 37 months (range, 12-49 months). Tolerance to dynamically locked nails was significantly better, with only one patient in Group B having cortical hypertrophy of the femur at the level of the tip of the nail, compared with six patients in Group A. Other outcomes were equal in both groups. Late tolerance to this new dynamically locked intramedullary hip screw is good, while retaining the known advantages of nailing of these fractures. PMID- 12579018 TI - Oblique osteotomy for multiplanar correction of malunions of the femoral shaft. AB - Between 1990 and 1996, six patients had oblique femoral osteotomies for symptomatic posttraumatic femoral malunions. In each patient, an oblique osteotomy was done that allowed for correction in the coronal, transverse, and sagittal planes. Fixation was achieved with lag screws supplemented by a neutralization plate. The average followup was 49 months (range, 18-99 months). All osteotomies healed at an average of 2.9 months (range, 2.5-4.0 months). Postoperatively, all patients had full range of motion in the ipsilateral hip and knee and could bear full weight without pain. Radiographically, the average deformity measured on anteroposterior radiographs improved from 21.6 degrees to 4.2 degrees varus. The average deformity on lateral radiographs improved from 22.5 degrees to 7.0 degrees antecurvatum. In all patients, the leg lengths were within 0.5 cm of each other and axial alignment was within 10 degrees of the contralateral limb. The use of an oblique osteotomy is a simple and predictable method of correcting complex posttraumatic femoral malunions and provides excellent surgical and clinical results. PMID- 12579019 TI - Locked nailing of severely comminuted or segmental humeral fractures. AB - Some comminuted or segmental humeral fractures have been well-treated by functional bracing, but some also have been reported to have a bad outcome. If surgery is required, unlocked nailing has the disadvantage of unstable fixation, and plating has the disadvantage of extensive soft tissue injury. In the current study, closed locked nailing was used to treat 23 Orthopaedic Trauma Association Type C humeral fractures (C1, five; C2, three; C3, 15). The patients were 12 men and 11 women with a mean age of 42.4 years. There were 16 closed fractures and four Type I, two Type II, and one Type IIIA open fractures. Eight fractures were in the proximal third, seven in the middle third, and eight in the distal third. With one nailing, 21 of 23 (91%) fractures achieved solid union in an average of 13.2 weeks. The two nonunions eventually united after another revision nailing and bone grafting. No patients had significant impairment of elbow function. Excellent or satisfactory shoulder function was obtained in 20 patients. Closed locked nailing showed reliable treatment results for severely comminuted or segmental humeral fractures. The keys to success include compression of the fractures, static locking, postoperative external support, and experienced surgical technique. PMID- 12579020 TI - Exoneration of NF-kappaB dysregulation in fibrodysplasia ossificans progressiva. AB - Fibrodysplasia ossificans progressiva is a disabling genetic disorder characterized by congenital skeletal malformations and progressive heterotopic ossification. New episodes of ossification are heralded by preosseous inflammatory lesions replete with B and T lymphocytes that overexpress bone morphogenetic protein-4. NF-kappaB is an inflammatory mediator that plays a critical role in developmental skeletogenesis and in suppression of bone morphogenetic protein-4 expression. Because of its multiple roles in inflammation, skeletogenesis, and bone morphogenetic protein-4 regulation, NF kappaB may play an important functional role in the pathogenesis of fibrodysplasia ossificans progressiva. To clarify the potential role of NF-kappaB in the pathophysiologic features of fibrodysplasia ossificans progressiva, the role of NF-kappaB in regulating bone morphogenetic protein-4 signaling in patient derived lymphoblastoid cell lines was examined. General NF-kappaB activity and specific NF-kappaB suppression of bone morphogenetic protein-4 expression were not altered in fibrodysplasia ossificans progressiva. In addition, despite the proximity of the gene for the p50 subunit of NF-kappaB (NFKB1 on long arm of chromosome 4) to the recently mapped locus for fibrodysplasia ossificans progressiva, a detailed linkage exclusion analysis in four multigenerational families with the disorder excluded NFKB1 as the causative gene for fibrodysplasia ossificans progressiva. These data exonerate NF-kappaB as the critical molecular and genetic pathogenic mediator in fibrodysplasia ossificans progressiva and, therefore, implicate a defect in another regulatory pathway as the cause for bone morphogenetic protein-4 overexpression in the disease. PMID- 12579021 TI - Ischiogluteal bursitis: multimodality imaging findings. AB - Ischiogluteal bursitis is a rare, infrequently recognized condition. Two cases of ischiogluteal bursitis in an 80-year-old man and a 75-year-old woman are presented. Both patients presented with a painful buttock mass. Contrast-enhanced multiplanar reconstruction computed tomography scans revealed a lesion with peripheral enhancement inferior to the ischial tuberosity. Magnetic resonance imaging revealed a lesion with low or intermediate signal intensity on T1 weighted images. T2-weighted images revealed a lesion with high signal intensity. Postcontrast magnetic resonance imaging scans revealed peripheral enhancement of the lesion. The multimodality imaging findings of ischiogluteal bursitis have not been described previously. Ischiogluteal bursitis should be considered in the differential diagnosis of a buttock mass. PMID- 12579022 TI - Immediate versus delayed meniscal allograft transplantation in sheep. AB - The aim of the current study was to assess the effect of immediate and delayed transplantation on articular cartilage. Icelandic sheep had surgery on the medial menisci in both knees. The knees were divided into four groups; 16 control knees had sham surgery, 15 knees had meniscectomy, 14 knees had immediate allograft transplantation, and 14 knees had delayed transplantation 3 months after meniscectomy. The two horns of the fresh allograft were fixed with suture anchors through bone tunnels. The sheep had autopsies 6 months after surgery. An additional six knees were examined 3 months after meniscectomy to examine the cartilage before the delayed implantation. Four standardized samples were taken from the medial compartment of the knee for histologic evaluation according to a modification of Mankin's system, where degenerative changes to the articular cartilage were scored semiquantitatively for structure, proteoglycan content, cellularity, and cell cloning. Knees that had meniscectomy and delayed transplantation showed equally high scores in articular cartilage degeneration, and both scored higher than knees that had immediate transplantation, which again scored higher than the control knees. In short-term observations, immediate medial meniscal allograft transplantation reduced but did not prevent degeneration of the articular cartilage. Immediate transplantation was superior to delayed transplantation in preventing cartilage degeneration. PMID- 12579023 TI - Response of a calcium sulfate bone graft substitute in a confined cancellous defect. AB - Calcium sulfate, plaster of Paris, has a long clinical history for use as a bone graft substitute in various skeletal sites. The current authors examined the in vivo response of calcium sulfate pellets alone or in combination with autogenous bone graft in a bilateral critical-size distal femoral cancellous defect in an adult sheep model. New thick bone formation was seen in defects filled with calcium sulfate pellets alone. Increased immunostaining for bone morphogenetic protein-2, bone morphogenetic protein-7, transforming growth factor-beta, and platelet derived growth factor was seen in defects filled with calcium sulfate pellets alone and in combination with autograft. The local acidity during calcium sulfate resorption is proposed as a possible in vivo mechanism for this type of material. PMID- 12579024 TI - Substance P and prostaglandin E2 release after shock wave application to the rabbit femur. AB - The biologic action of extracorporeal shock wave application on the musculoskeletal system is poorly understood. To prove the hypothesis that alterations of tissue concentrations of substance P and prostaglandin E(2) are involved in the biologic action of shock waves, extracorporeal shock waves with energy flux density of 0.9 mJ/mm2 (1500 pulses at 1/second) were applied in vivo to the distal femur of rabbits. The concentrations of substance P and prostaglandin E(2) eluted from the periosteum of the femur were measured. Compared with the untreated contralateral hindlimbs, substance P release from the periosteum from the femur was increased 6 hours and 24 hours after extracorporeal shock wave application, but was decreased 6 weeks after extracorporeal shock wave application. By contrast, extracorporeal shock wave application did not result in altered prostaglandin E(2) release from the periosteum from the femur. Remarkably, there was a close relationship between the time course of substance P release found here, and the well-known clinical time course of initial pain occurrence and subsequent pain relief after extracorporeal shock wave application to tendon diseases. Accordingly, substance P might be involved in the biologic action of extracorporeal shock wave application on tissue of the musculoskeletal system. This is the first study providing insights into the molecular mechanisms of extracorporeal shock wave application to the musculoskeletal system. PMID- 12579025 TI - Bacterial skin contamination after surgical preparation in foot and ankle surgery. AB - An effective presurgical preparation is an important step in limiting surgical wound contamination and preventing infection. The purpose of this study was to evaluate residual bacterial skin contamination after surgical skin preparation in foot and ankle surgery to determine if current techniques are satisfactory in eliminating harmful pathogens. Fifty consecutive patients having surgical procedures of the foot and ankle were studied. Each lower extremity was prepared randomly with either a one-step povidone-iodine topical gel or a two-step iodophor scrub followed by a povidone-iodine paint. After preparation and draping, cultures were obtained at three locations: the hallux nailfold, web space between the second and third, and fourth and fifth toes, and the anterior ankle (control). In the gel group, positive cultures were obtained from 76% of halluces, 68% of toes, and 16% of controls. In the scrub and paint group, positive cultures were obtained from 84% of halluces, 76% of toes, and 28% of controls. Numerous pathogens were cultured, with Staphylococcus epidermidis being the most prevalent. Based on the findings of the current study, presurgical skin preparation with a povidone-iodine based topical bactericidal agent is not sufficient in eliminating pathogens in foot and ankle surgery. The unique environment of the foot and its resident organisms may play a role in the higher infection rates associated with surgery of the foot and ankle. PMID- 12579026 TI - Hydroxyapatite composite resin cement augmentation of pedicle screw fixation. AB - Pedicle screw stability is poor in osteopenic vertebrae attributable, in part, to low screw-bone interface strength. The current authors examined cement augmentation using a low curing temperature hydroxyapatite and bis-phenol-A glycidol methacrylate-based composite resin. This cement may stiffen the screw bone interface and reduce the harmful effects associated with polymethylmethacrylate regarding temperature and toxic monomer. Thirty-five lumbar vertebrae from human cadavers were instrumented with pedicle screws, with one pedicle previously injected with cement and the other as the control. Caudocephalad toggling of +/- 1 mm for 1600 cycles was applied to the pedicle screws, and the resulting forces supported by the implant-bone interface were captured by a load cell. A curve was constructed from the peak caudal load for each cycle and three mechanical measures parameterized this curve: (1) initial load; (2) rate of load decay during the first 400 cycles; and (3) final load. The initial load increased by 16% as a result of cement augmentation, the final load increased by 65%, and the rate of load decay decreased by 59%. Cement augmentation of pedicle screws increased the stiffness and stability of the screw bone interface. PMID- 12579027 TI - Articular cartilage restoration with costal cartilage previously fused with bone. AB - A novel procedure was developed for restoration of an articular cartilage defect using an autologous costal cartilage prepared with iliac bone, and the durability in vivo of this biologic construct was examined. First, an osteochondral complex was prepared (successful preparation, 67 of 80). Cancellous bone blocks isolated from the ilium of male Japanese White rabbits aged 5 months were implanted onto the surface of the costal cartilage before being tied by a pair of 3-0 silk thread sutures that were looped around the costal cartilage from behind. Second, 3 months later, the bone-attached costal cartilage was harvested and implanted into a full-thickness cartilage defect induced in a trochlear groove of the femur. All of the grafts were fixed to the recipient, maintaining its cartilage structure until 6 months (n = 28) and 12 months (n = 12) after implantation. However, when the costal cartilage without any bony portion was implanted into a similarly induced defect, 42% (10 of 24) were detached from the recipient before 12 months after implantation. The nontreated defect did not heal spontaneously to a satisfactory level (n = 12). These findings suggest that an osteochondral fragment, prepared by grafting cancellous bone onto costal cartilage, can be used for articular cartilage restoration. PMID- 12579028 TI - Assessment of external fixator reusability using load- and cycle-dependent tests. AB - No standard method has been established for investigating repeated use of an external fixator. The purpose of the current study was to establish a fatigue testing method for assessing fixator frame reuse. A unilateral DynaFix trade mark external fixator system was tested using high-load and low-cycle (900-150 N at 5 Hz) and low-load and high-cycle (450-100 N at 10 Hz) tests (assumed one use of 500,000 and 1 million cycles, respectively). These loading conditions were selected to simulate single clinical use and to satisfy Food and Drug Administration requirements. In the high-load low-cycle test, substantial failure of the serrated joint occurred before completion of the first simulated use. In the low-load high-cycle test, all fixators completed three simulated clinical uses without failure, although (1/4) of the serrated joint components had hairline cracks. The high-load low-cycle test identified the fixator components which should be examined and replaced if reuse of the fixator is to be considered. Wear and deformation of the set screw on the rotary joint and telescoping mechanisms were observed in the low-load high-cycle test but not in the high-load low-cycle test. Therefore, if the unilateral DynaFix trade mark fixators are being considered for reusability, the number of reuses should be limited as the whole structure of the device will experience fatigue damage as the loading cycle increases. PMID- 12579030 TI - A symposium that salutes volunteers in orthopaedics. PMID- 12579029 TI - Metal-on-metal versus polyethylene in hip arthroplasty: a randomized clinical trial. AB - A prospective, randomized, blinded clinical trial was done to evaluate polyethylene versus metal bearing surfaces in total hip replacement. Forty-one patients were randomized to receive either a metal (23 patients) or a polyethylene (18 patients) insert. The femoral and acetabular components were identical with the acetabular insert the only variable. Patients were assessed preoperatively and postoperatively using radiographs, multiple outcome measures (Western Ontario MacMaster University Score, Harris hip score, Short Form-12), erythrocyte metal ion analysis (cobalt, chromium, titanium), and urine metal ion analysis (cobalt, chromium, titanium). Patients were followed up for a minimum of 2 years (mean 3.2 years; range, 2.2-3.9 years). There were no differences in radiographic outcomes or outcome measurement tools between patients. Patients receiving a metal-on-metal articulation had significantly elevated erythrocyte and urine metal ions compared with patients receiving a polyethylene insert. Patients who had metal-on-metal inserts had on average a 7.9-fold increase in erythrocyte cobalt, a 2.3-fold increase in erythrocyte chromium, a 1.7-fold increase in erythrocyte titanium, a 35.1-fold increase in urine cobalt, a 17.4 fold increase in urine chromium, and a 2.6-fold increase in urine titanium at 2 years followup. Patients receiving a polyethylene insert had no change in erythrocyte titanium, urine cobalt, or urine chromium and a 1.5-fold increase in erythrocyte cobalt, a 2.2-fold increase in erythrocyte chromium, and a 4.2-fold increase in urine titanium. Forty-one percent of patients receiving metal-on metal articulations had increasing metal ion levels at the latest followup. PMID- 12579031 TI - Leg mass in a 61-year-old man. PMID- 12579032 TI - Ankle tumor in a 5-year-old boy. PMID- 12579033 TI - Knee pain in a 13-year-old boy. PMID- 12579034 TI - An emerging role for nitric oxide in sickle cell disease vascular homeostasis and therapy. AB - Nitric oxide participates in the compensatory response to chronic vascular injury in patients with sickle cell disease. The authors have found reductions of basal and stimulated nitric oxide production and responses to exogenous nitric oxide in male patients with sickle cell disease. Gender differences in nitric oxide bioavailability are probably caused in part by the protective effects of ovarian estrogen on nitric oxide synthase expression and activity in women. Further, in men, and likely all patients during vaso-occlusive crisis and the acute chest syndrome, nitric oxide is destroyed by increased circulating plasma hemoglobin and superoxide. The combined effects of inhaled nitric oxide gas of improving pulmonary ventilation to perfusion matching and hemodynamics, reducing alveolar and systemic inflammation, and inhibiting circulating plasma hemoglobin (and thus restoring peripheral nitric oxide bioavailability) may modulate the course of the disease, including the frequency and severity of vaso-occlusive crises and acute chest syndrome episodes. Possible effects of chronic nitric oxide-based therapies on erythrocyte density, pulmonary artery pressures, and fetal hemoglobin induction deserve study. PMID- 12579035 TI - Malaria and anemia. AB - Anemia due to infection is a major health problem in endemic areas for young children and pregnant women. The anemia is caused by excess removal of nonparasitized erythrocytes in addition to immune destruction of parasitized red cells, and impaired compensation for this loss by bone marrow dysfunction. The pathogenesis is complex, and a predominant mechanism has not been identified. Certain parasite and host characteristics may modify the anemia. Concomitant infections and nutritional deficiencies also contribute to anemia and may interact with the malarial infection. Few preventive strategies exist, and the management of severe malarial anemia with blood transfusion carries a risk of HIV transmission. The current increase in malaria-specific childhood mortality in sub Saharan Africa attributed to drug-resistant infection is likely partly related to an increase in severe anemia. This review summarizes recent findings on the pathogenesis and epidemiology of malarial anemia. PMID- 12579036 TI - Regulation of erythrocyte membrane protein gene expression. AB - Proteins of the erythrocyte membrane have served as the prototypes of homologous families of multifunctional proteins in erythroid and nonerythroid cells. These proteins demonstrate many different cell type, tissue-specific, and developmental stage-specific functions. This complex pattern of functional diversity appears to have evolved from the cell type, tissue-specific, developmentally regulated expression of multiple protein isoforms. Isoform diversity arises from different gene products from related genes; from differential, alternate splicing of the same gene product; from the use of tissue-specific promoters; and from alternate polyadenylation. The identification and characterization of the regulatory elements that control erythrocyte membrane protein gene expression have important implications for several biologic processes. These include disease pathogenesis, membrane assembly, hematopoiesis, gene regulation, and direction of other erythroid-specific genes in transgenic mouse and gene therapy applications. PMID- 12579037 TI - Hemoglobin switching: new insights. AB - During the past year, many interesting advances have been made regarding molecular mechanisms controlling beta-like globin gene switching. Throughout the beta locus, -acting elements exist that are dynamically bound by trans-acting proteins, including transcription factors, coactivators, repressors, and chromatin modifiers. Characterization of transcription factors, their interaction with one another, and an ever-increasing role for chromatin structure in gene expression have enhanced understanding of the mechanism of globin gene switching. The studies reviewed here contribute new insights on the interplay between acting elements, transcription factors, and chromatin modifiers that underlie globin gene switching during development. PMID- 12579039 TI - Vascular development: molecular logic for defining arteries and veins. AB - Vascular development is a complex process. In the past decade, significant advances have been made in the understanding of molecular mechanisms underlying vascular system development. Most recently, a model describing a signaling pathway that governs the differentiation of arteries and veins has emerged. Furthermore, the idea that angiogenesis of arteries and veins are coordinately but also differentially regulated has been proposed based on several pieces of molecular evidence. This article discusses the historical background leading to these most recent discoveries of signaling pathways that dictate arterial-venous formation and proposes a model that describes how arterial and venous systems are specified and established. The article also discusses future perspectives of this exciting area of vascular development in concluding remarks. PMID- 12579041 TI - Von Willebrand factor. AB - The adhesive protein von Willebrand factor contributes to platelet function by mediating the initiation and progression of thrombus formation at sites of vascular injury. In the last 2 years, there has been considerable progress in explaining the biologic properties of von Willebrand factor. The three dimensional structure of specific domains has been explained, with the demonstration of distinct conformational changes in the A1 domain caused by single amino acid substitutions associated with enhanced binding to platelets. The structural and functional properties of the interaction between the von Willebrand factor A1 domain and glycoprotein Ibalpha have also been elucidated in greater detail, bringing researchers closer to understanding how this adhesive bond can oppose the fluid dynamic effects of rapidly flowing blood to initiate thrombus formation and, concurrently, contribute to platelet activation. Because hemodynamic forces greatly influence platelet responses to vascular injury in stenosed and partially occluded arteries, a detailed description of how von Willebrand factor interacts with tissues and platelets may help in the design of more specific therapeutic inhibitors of arterial thrombosis. Moreover, enlightening findings have been obtained on the link between regulation of von Willebrand factor multimer size and microvascular thrombosis. This progress in basic research has provided critical information to define with greater precision the role of von Willebrand factor in vascular biology and pathology. PMID- 12579040 TI - Angiogenesis: vascular remodeling of the extracellular matrix involves metalloproteinases. AB - Endothelial cell invasion is an essential event during angiogenesis (the formation of new blood vessels). This process involves the degradation of the extracellular matrix, the basement membrane, and interstitial stroma, and is governed by the activation of matrix metalloproteinases. However, the contribution of matrix metalloproteinases in angiogenesis is much more complicated. Tumor growth above a certain size is dependent on new vessels. A number of studies have demonstrated that treating tumors with matrix metalloproteinase inhibitors results in tumor reduction and a decrease in tumor angiogenesis. Matrix metalloproteinases as sole matrix eaters or degraders is a matter of the past. Not only tumor cells but more importantly bystander cells such as stromal cells produce matrix metalloproteinases. Matrix metalloproteinases therefore are also part of the pathologic microenvironment in different diseases. This enzymatic microenvironment dictates the endothelial cell fate, the angiogenic switch, and finally angiogenesis. During recent years, the role of matrix metalloproteinases has expanded, and their function as modulators of biologically active signaling molecules has drawn much attention. Depending on their substrate (growth factors or their receptors, extracellular matrix components, and angiogenic factors), matrix metalloproteinase activation results in the generation of proangiogenic or antiangiogenic factors. These data challenge the old concept that matrix metalloproteinases are simply proangiogenic. The knowledge of the local enzymatic profile and what, where, and how matrix metalloproteinases are involved in angiogenesis of tumors or other diseases will help design future therapeutic strategies better reflecting the complexity of the underlying biologic process of angiogenesis. PMID- 12579042 TI - Cell-cell interactions: leukocyte-endothelial interactions. AB - Interactions of leukocytes with endothelial cells are early events in acute and chronic inflammation, immune surveillance of tissues, and wound defense and repair. In contrast with their requisite roles in host defense, dysregulated leukocyte-endothelial interactions mediate inflammatory tissue injury, thrombosis, and other pathologic sequelae. Recent observations also identify dysregulated leukocyte-endothelial interactions in neoplasia and sickle cell vasculopathy. Leukocyte interactions with inflamed endothelial cells are mediated by selectins, signaling molecules that include lipids and chemokines, integrins and their ligands, and junctional molecules. They provide multiple checkpoints for regulation in physiologic inflammation and hemostasis and for dysregulation in pathologic syndromes. Neutrophil-endothelial encounters illustrate a multistep paradigm for inflammatory cell-cell interactions and provide the basis for multiple variations on the central themes. PMID- 12579043 TI - Post-hemorrhagic shock mesenteric lymph activates human pulmonary microvascular endothelium for in vitro neutrophil-mediated injury: the role of intercellular adhesion molecule-1. AB - BACKGROUND: Splanchnic hypoperfusion is believed to be central in the pathogenesis of hemorrhagic shock-induced acute respiratory distress syndrome and multiple organ failure. Our previous work focused on the portal circulation as the conduit for gut-derived mediators of acute respiratory distress syndrome. Our current focus is the proinflammatory effects of postshock mesenteric lymph. We hypothesize that postshock lymph induces neutrophil (PMN)-mediated endothelial cell damage in an intercellular adhesion molecule-1 (ICAM-1)-dependent fashion, and devised a two-insult model to test this hypothesis. METHODS: Rats (n > or = 5) underwent hemorrhagic shock (mean arterial pressure, 40 mm Hg for 30 minutes) and resuscitation (shed blood plus two times crystalloid) with lymph collection. Human pulmonary microvascular endothelial cells (HMVECs) were divided into three groups and grown to near confluence. Group 1 was incubated for 6 hours in 1% preshock or postshock lymph and ICAM-1 was measured by flow cytometry. Group 2 consisted of coculture of HMVECs and PMNs after endothelial cell activation to determine whether postshock lymph would stimulate PMN adherence. Group 3 was incubated under identical conditions, but PMNs were added for 30 minutes, and then activated with 4.5 micromol/L lysophosphatidylcholine (lyso-PC) for 1 hour to ascertain cytotoxicity. HMVEC density was measured using microscopy and recorded as HMVECs per millimeter squared. ICAM-1-blocking antibody and isotype control were used to assess the effects of ICAM-1 on PMN cytotoxicity. A buffer control was used for comparison using analysis of variance with Tukey's correction. RESULTS: Postshock lymph activated HMVECs for increased surface expression of ICAM-1 and stimulated PMNs to adhere to endothelial cell monolayers. Activation of PMNs with lyso-PC in the presence of postshock lymph resulted in marked HMVEC death. The addition of an ICAM-1-blocking antibody abrogated this effect. Neither postshock lymph alone (758 +/- 35 HMVECs/mm(2)), nor postshock lymph in the presence of quiescent PMNs alone (734 +/- 28 HMVECs/mm(2)), nor lymph plus lyso-PC (834 +/- 21 HMVECs/mm(2)) provoked endothelial cell damage. CONCLUSION: Postshock mesenteric lymph activates endothelial cells for increased ICAM-1 expression and PMN adherence. Furthermore, postshock lymph acts as an inciting event in a two-event in vitro model of PMN mediated endothelial cell injury. These findings further substantiate the key mechanistic role of mesenteric lymph in hemorrhagic shock-induced acute lung injury and suggest that ICAM-1 expression is pivotal in the two-event model of multiple organ failure. PMID- 12579044 TI - Venous thromboembolism in trauma: a local manifestation of systemic hypercoagulability? AB - BACKGROUND: The purpose of this study was to evaluate the relative importance of systemic hypercoagulability, preexisting and acquired risk factors, and specific injury patterns in the development of venous thromboembolism (VTE) after injury. METHODS: Injured patients with an Injury Severity Score > or = 15 were followed with lower extremity venous duplex ultrasonography, prothrombin fragment 1 + 2, and quantitative D-dimer levels at 1 and 3 days and then weekly until discharge. RESULTS: Among 101 patients with a mean Injury Severity Score of 27.3 +/- 10.5 followed for 12.4 +/- 8.7 days, 28 (27.7%) developed a lower extremity thrombosis, 2 (1.9%) sustained a pulmonary embolism, and 1 (0.9%) had a symptomatic upper extremity thrombosis. Although admission fragment 1 + 2 and D dimer levels were elevated in 81.4% and 100% of patients, respectively, mean levels were not significantly different in those with or without VTE. VTE was more common (p < 0.05) among those with obesity, age > 40 years, immobilization for > 3 days, spine fractures, and lower extremity fractures. However, only obesity (p = 0.004) and immobilization > 3 days (p = 0.05) were independent predictors of VTE in a multivariate analysis. CONCLUSION: Although elevated in seriously injured patients, neither markers of activated coagulation nor specific injury patterns are predictive of VTE. Associations with immobilization and obesity suggest that VTE after injury is a systemic hypercoagulable disorder with local manifestations of thrombosis related to lower extremity stasis. PMID- 12579045 TI - Expedited discharge in trauma patients requiring anticoagulation for deep venous thrombosis prophylaxis: the LEAP Program. AB - OBJECTIVE: With rising health care costs, methods to decrease length of hospital stay without compromising care are necessary. One area that extends length of stay in trauma patients is inpatient anticoagulation to a therapeutic international normalized ratio. The 1998 American College of Chest Physicians guidelines recommend thromboprophylaxis with low-molecular-weight heparin (LMWH) and oral warfarin in this population. The LMWH Expedited Anticoagulation Program (LEAP) was created with the following goals: to decrease the number of inpatient warfarin days and to reduce overall number of hospital days. METHODS: Inpatient anticoagulation was initiated with warfarin and LMWH. LEAP included early multidisciplinary collaboration to ensure third-party approval, outpatient primary care physician follow-up, and LMWH self-injection before discharge. Patients were discharged on LMWH (discontinued by primary care provider when a therapeutic international normalized ratio was attained) and warfarin (continued until resolution of orthopedic injuries). From August 2000 to August 2001, adult patients were included in the prospective study. Primary inclusion criteria were blunt acetabular fracture, bilateral lower extremity fracture, and contralateral upper and lower extremity fractures. Patients with similar injuries receiving warfarin for deep venous thrombosis prophylaxis between June 1999 and June 2000 were the control population. Anticoagulation care was similar for the study and control subjects. RESULTS: There were 182 patients evaluated for LEAP inclusion. After initial evaluation, 108 patients were enrolled in LEAP (Injury Severity Score of 13). There were 69 control subjects (Injury Severity Score of 13). The average number of inpatient warfarin days was decreased from 8.8 days to 5.0 days in the control and study populations, respectively (p < 0.0001). The average length of hospitalization was shortened from 17.3 days in the control group to 12.9 days in the study (LEAP) population (p < 0.002). CONCLUSION: LEAP has successfully decreased the number of inpatient days on warfarin and total hospital days for trauma patients requiring deep venous thrombosis prophylaxis. These results have substantially decreased health care costs and increased available hospital beds in this era of high hospital occupancy. PMID- 12579046 TI - Reliability of mixed venous oxygen saturation as an indicator of the oxygen extraction ratio demonstrated by a large patient data set. AB - BACKGROUND Mixed venous saturation (SVO2) represents the oxygen extraction ratio (O2ER) through the following relationship: SVO2 approximately equal 1 - O2ER, assuming a negligible contribution of dissolved oxygen to blood oxygen content. We sought to determine whether altered FiO2 levels would affect the correlation between SVO2 and O2ER. METHODS: A theoretical analysis was performed to determine the extent to which SVO2 should be affected by different Fio2 levels at various ranges of O2ER using several linked spreadsheets. The relationships obtained were compared with actual patient data stored in a large patient data set. RESULTS Theoretical analysis indicated that increases in Fio2 should produce a small stepwise increase in the observed SVO2 that is independent of O2ER. However, analysis of 8,324 simultaneous data points for SVO2 and O2ER failed to demonstrate this pattern, primarily because patients with higher Fio2 concentrations tend to have higher O2ERs, and hence lower SVO2. CONCLUSION SVO2 measurements are not significantly affected by Fio2 levels in critically ill patients. PMID- 12579047 TI - Tissue oxygen monitoring during hemorrhagic shock and resuscitation: a comparison of lactated Ringer's solution, hypertonic saline dextran, and HBOC-201. AB - BACKGROUND: The ideal resuscitation fluid for the trauma patient would be readily available to prehospital personnel, universally compatible, effective when given in small volumes, and capable of reversing tissue hypoxia in critical organ beds. Recently developed hemoglobin-based oxygen-carrying solutions possess many of these properties, but their ability to restore tissue oxygen after hemorrhagic shock has not been established. We postulated that a small-volume resuscitation with HBOC-201 (Biopure) would be more effective than either lactated Ringer's (LR) solution or hypertonic saline dextran (HSD) in restoring baseline tissue oxygen tension levels in selected tissue beds after hemorrhagic shock. We further hypothesized that changes in tissue oxygen tension measurements in the deltoid muscle would reflect the changes seen in the liver and could thus be used as a monitor of splanchnic resuscitation. METHODS: This study was a prospective, blinded, randomized resuscitation protocol using anesthetized swine (n = 30), and was modeled to approximate an urban prehospital clinical time course. After instrumentation and splenectomy, polarographic tissue oxygen probes were placed into the liver (liver PO2) and deltoid muscle (muscle PO2) for continuous tissue oxygen monitoring. Swine were hemorrhaged to a mean arterial pressure (MAP) of 40 mm Hg over 20 minutes, shock was maintained for another 20 minutes, and then 100% oxygen was administered. Animals were then randomized to receive one of three solutions: LR (12 mL/kg), HSD (4 mL/kg), or HBOC-201 (6 mL/kg). Physiologic variables were monitored continuously during all phases of the experiment and for 2 hours postresuscitation. RESULTS: At a MAP of 40 mm Hg, tissue PO2 was 20 mm Hg or less in both the liver and muscle beds. There were no significant differences in measured liver or muscle PO2 values after resuscitation with any of the three solutions in this model of hemorrhagic shock. When comparing the hemodynamic effects of resuscitation, the cardiac output was increased from shock values in all three animal groups with resuscitation, but was significantly higher in the animals resuscitated with HSD. Similarly, MAP was increased by all solutions during resuscitation, but remained significantly below baseline except in the group of animals receiving HBOC-201 (p < 0.01). HBOC-201 was most effective in both restoring and sustaining MAP and systolic blood pressure. There was excellent correlation between liver and deltoid muscle tissue oxygen values (r = 0.8, p < 0.0001). CONCLUSION: HBOC-201 can be administered safely in small doses and compared favorably to resuscitation with HSD and LR solution in this prehospital model of hemorrhagic shock. HBOC-201 is significantly more effective than HSD and LR solution in restoring MAP and systolic blood pressure to normal values. Deltoid muscle PO2 reflects liver PO2 and thus may serve as an index of the adequacy of resuscitation in critical tissue beds. PMID- 12579048 TI - Enhanced expression of intranuclear NF-kappa B in primed polymorphonuclear leukocytes in systemic inflammatory response syndrome patients. AB - BACKGROUND: Nuclear factor-kappa B (NF-kappa B) plays a critical role in the cellular response to a variety of stimuli, and it regulates the production of various inflammatory cytokines, adhesion molecules, and enzymes. Polymorphonuclear leukocytes (PMNLs) play a central role in systemic inflammatory response after severe insult. The role of NF-kappa B in activation of PMNLs, however, has not been clear. We developed a simple flow cytometric method for quantifying expression of intranuclear NF-kappa B in PMNLs, and we used it to evaluate NF-kappa B activity in patients with systemic inflammatory response syndrome (SIRS). METHODS: Thirty patients who fulfilled the criteria for SIRS and 24 healthy volunteers were included as study subjects. Expression of intranuclear NF-kappa B with and without stimulation by lipopolysaccharide was quantified by our new method. Oxidative activity in PMNLs with and without formylmethionyl leucyl-phenylalanine stimulation was measured by flow cytometry. Levels of interleukin-6, interleukin-8, PMNL elastase, and nitric oxide metabolites in blood were also measured. RESULTS: Expression of intranuclear NF-kappa B in PMNLs both with and without LPS stimulation was significantly elevated in SIRS patients in comparison with that of healthy volunteers. PMNL oxidative activity was significantly elevated in SIRS patients. Positive correlation was observed between intranuclear NF-kappa B expression and PMNL oxidative activity, whereas no relation was observed between intranuclear NF-kappa B expression and serum concentrations of chemical mediators. CONCLUSION: Our new flow cytometric method proved useful for quantifying intranuclear NF-kappa B expression in PMNLs. In PMNLs from SIRS patients, intranuclear NF-kappa B expression and oxidative activity were significantly elevated with positive correlation, and enhanced expression of NF-kappa B may play an important role in PMNL activation in SIRS. PMID- 12579049 TI - Relationship between age and lower extremity fractures in frontal motor vehicle collisions. AB - BACKGROUND: Older adults (aged > or = 65 years) represent the single fastest growing segment of the United States population and will comprise one in five Americans during the third decade of this century. As this population segment rapidly expands, lower extremity fractures (LE Fx) and their associated disability will become a greater public health concern. The purpose of this study was to quantify the risk for LE Fx from motor vehicle collisions (MVCs) according to age. METHODS: The 1995 to 2000 National Automotive Sampling System data files were used. Study entry was limited to front-seat occupants involved in frontal MVCs. Risk ratios for LE Fx and age were adjusted for gender, driver versus passenger, seat belt use, airbag deployment, delta-V, intrusion, and vehicle type. RESULTS: Beginning in the fourth decade, there was a trend of higher relative risk for LE Fx with age that reached statistical significance in the seventh decade of life. CONCLUSION: This study documented an increased risk of LE Fx in older MVC occupants. Efforts to prevent these disabling injuries and to better protect occupants' lower extremities in MVCs should include improved vehicle design and reevaluation of the existing federal motor vehicle safety standards. PMID- 12579050 TI - Effect of airbag deployment on head injuries in severe passenger motor vehicle crashes in Ontario, Canada. AB - BACKGROUND: The purpose of this study was to identify and analyze factors contributing to both airbag deployment and resulting head injuries (HIs) and to quantify the effect of airbag deployment on head injuries, in terms of odds of head injury and severity, in severe motor vehicle collisions (MVCs). METHODS: Data were derived from severely injured (Injury Severity Score [ISS] > 12) drivers treated at Ontario's lead trauma hospitals (n = 1,272), and included all MVC driver deaths in the province (n = 665) from 1997-98. We conducted an epidemiologic description and a case-control study to compare drivers with and without HIs. Statistical analysis included Pearson's chi2, Wilcoxon rank-sum, and multiple logistic regression tests. RESULTS: Seventy-one percent of drivers were men, peaking in the 25- to 34-year age group. The most common impact involved multiple vehicles (62%) approaching each other. Overall, 59% of crashes had a frontal location of impact. HIs were significantly associated with a lower age (median, 36 vs. 43 years), seat belt use (53% vs. 59%), and airbag deployment (7% vs. 10%), with higher ISS (median, 34 vs. 22), ejection (20% vs. 10%), and mortality rate (44% vs. 35%). Airbag deployment was associated with higher age and seat belt use, and lower ISS, ejection, and deaths. Importantly, there were fewer HIs with the deployment of an airbag (64% vs. 73%) and a lower severity of HI. When logistic regression was used to control for the effects of possible confounders, airbag deployment was not statistically associated with one's odds of HI (odds ratio, 0.827; 95% confidence interval, 0.560-1.220), but ISS, age, and ejection were. CONCLUSION: Airbag deployment did not significantly lower a driver's odds of head injury in a severe MVC, but it did significantly lower the severity of head injury. This is a significant finding, given that 72% of our study population sustained a head injury and the importance of lowering the severity of these head injuries in terms of patients' ultimate outcome. The most important factor associated with head injuries was ejection, which nearly doubled a driver's odds of head injury (odds ratio, 1.759; 95% confidence interval, 1.201 2.577). This reinforces the supplementary protective effect of an airbag and that "buckling up" and keeping occupants in the vehicle is of primary importance in the prevention of head injuries. PMID- 12579051 TI - Motor vehicle crash-related mortality is associated with prehospital and hospital based resource availability. AB - BACKGROUND: To date, attempts to assess the relationship between motor vehicle collision (MVC)-related mortality and medical resources availability have largely been unsuccessful. METHODS: Information regarding sociodemographic characteristics, prehospital resources, and hospital-based resources for each county (n = 67) in the state of Alabama was obtained. MVC-related mortality rates (deaths per 1,000 collisions) by county were calculated and compared according to prehospital and hospital-based resource availability within each county after correcting for sociodemographic factors. RESULTS: Counties with 24-hour availability of a general surgeon, orthopedic surgeon, neurosurgeon, computed tomographic scanner, and operating room were shown to have decreased MVC-related mortality (relative risk [RR], 0.88). The same was true for those counties with hospitals classified as Level I-II (RR, 0.71) and Level III-IV (RR, 0.83) trauma centers compared with counties with no trauma centers. CONCLUSION: Appropriate, readily available hospital-based resources are associated with lower MVC-related mortality rates. This information may be useful in trauma system planning and development. PMID- 12579052 TI - Comparison of 10 different hemostatic dressings in an aortic injury. AB - BACKGROUND: Uncontrolled hemorrhage is the leading preventable cause of death on the battlefield. Similarly, hemorrhage accounts for 80% of all deaths within the first 48 hours of injury in civilian trauma patients. New methods of hemostasis are required to reduce hemorrhagic mortality. The purpose of this study was to compare nine hemostatic dressings for their efficacy in controlling bleeding from an otherwise fatal aortic injury in a pig model. Each hemostatic dressing was compared with the current standard U.S. Army field gauze dressing for a 1-hour period. METHODS: Fifty-nine anesthetized pigs were instrumented with catheters and splenectomized. Nine test dressings (n = 5 per group) and two control groups (gauze, n = 9; suture, n = 5) were applied to a 4.4-mm aortotomy through the spraying jet of blood, and direct pressure was held for 4 minutes and then released. Survival, blood loss, and other variables were measured over a 1-hour period. RESULTS: All animals with fibrin dressing and those receiving suture repair (five of five in both groups) survived the 1-hour observation period with minimal bleeding in the postocclusion period (< 37 mL). Those in the other dressing groups exsanguinated within 10 minutes, except for two animals in the gauze group surviving 1 hour. CONCLUSION: With one 4-minute application, a single fibrin dressing stopped bleeding from an aortotomy, which was equivalent to sutured repair. No other test group exhibited any evidence of significant hemostatic efficacy. PMID- 12579053 TI - Blunt cardiac injuries that require operative intervention: an unsuspected injury. PMID- 12579054 TI - Incidence of hollow viscus injury in blunt trauma: an analysis from 275,557 trauma admissions from the East multi-institutional trial. AB - BACKGROUND: Blunt hollow viscus injury (HVI) is uncommon. No sufficiently large series has studied the prevalence of these injuries in blunt trauma patients. This study defines the prevalence of blunt HVI, in addition to the associated morbidity and mortality rates for this diagnosis on the basis of a series of over 275,000 trauma admissions. METHODS: Patients with blunt small bowel injury (SBI) were identified from the registries of 95 trauma centers for a 2-year period (1998-1999). Each HVI patient (case) was matched by age and Injury Severity Score with a blunt trauma patient receiving an abdominal workup who did not have HVI (control). Patient level data were abstracted by individual chart review. Institution level data were collected on total numbers for trauma admission demographics and on total diagnostic examinations performed. RESULTS: From 275,557 trauma admissions, 227,972 blunt injury patients were identified. HVI was rare, with 2,632 patients identified from this group. Perforating small bowel injury accounted for less than 0.3% of blunt admissions. Mortality and morbidity were high for HVI. Controlling for injury severity, patients with HVI were usually at higher risk of death than non-HVI patients. CONCLUSION: HVI is a rare but deadly phenomenon. The high mortality rates reflect the severity of the HVI and associated injuries. HVI patients should be carefully monitored for related injuries and complications. PMID- 12579055 TI - Current diagnostic approaches lack sensitivity in the diagnosis of perforated blunt small bowel injury: analysis from 275,557 trauma admissions from the EAST multi-institutional HVI trial. AB - OBJECTIVES: Blunt SBI is infrequent and its diagnosis may be difficult, especially in the face of confounding variables. The purpose of this study was to evaluate methods for making the diagnosis of blunt SBI. METHODS: Patients with blunt small bowel injury (SBI) were identified from the registries of 95 trauma centers for a 2-year period (1998-1999). Patients with SBI (cases) were matched by age and Injury Severity Score with a blunt trauma patient receiving an abdominal workup who did not have SBI (controls). RESULTS: Logistic regression models were unable to differentiate SBI with perforation from SBI without perforation. Thirteen percent of patients with documented perforating SBI had normal abdominal computed tomographic scans preoperatively. CONCLUSION: Alone or in combination, current diagnostic approaches lack sensitivity in the diagnosis of perforated SBI. Improvements in diagnostic methods and approaches are needed to ensure the prompt diagnosis of this uncommon but potentially devastating injury. PMID- 12579056 TI - Endotracheal intubation in the field does not improve outcome in trauma patients who present without an acutely lethal traumatic brain injury. AB - OBJECTIVES: There is an absence of prospective data evaluating the impact of prehospital intubation in adult trauma patients. Our objectives were to determine the outcome of trauma patients intubated in the field who did not have an acutely lethal traumatic brain injury (death within 48 hours) compared with patients who were intubated immediately on arrival to the hospital. METHODS: Prospective data were collected on 191 consecutive patients admitted to the trauma center with a field Glasgow Coma Scale score < or = 8 and a head Abbreviated Injury Scale score > or = 3 who were either intubated in the field or intubated immediately at admission to the hospital. Patients who died within 48 hours of admission and transfers were excluded from the study. RESULTS: Of the 191 patients, 176 (92%) sustained blunt trauma and 25 (8%) were victims of penetrating trauma. Seventy eight (41%) of the 191 patients were intubated in the field and 113 (59%) were intubated immediately at admission. There was no significant difference in age, Glasgow Coma Scale score, head Abbreviated Injury Scale score, or Injury Severity Score between the two groups. Patients who were intubated in the field had a significantly higher morbidity (ventilator days, 14.7 vs. 10.4; hospital days, 20.2 vs. 16.7; and intensive care unit days, 15.2 vs. 11.7) compared with patients intubated on immediate arrival to the hospital and nearly double the mortality (23% vs. 12.4). Field-intubated patients had a 1.5 times greater risk of nosocomial pneumonia compared with hospital-intubated patients. CONCLUSION: Prehospital intubation is associated with a significant increase in morbidity and mortality in trauma patients with traumatic brain injury who are admitted to the hospital without an acutely lethal injury. A randomized, prospective study is warranted to confirm these results. PMID- 12579057 TI - Harbingers of poor outcome the day after severe brain injury: hypothermia, hypoxia, and hypoperfusion. AB - BACKGROUND: Traumatic brain injury (TBI) can be compounded by physiologic derangements that produce secondary brain injury. The purpose of this study is to elucidate the frequency with which physiologic factors that are associated with secondary brain injury occur in patients with severe closed head injuries and to determine the impact of these factors on outcome. METHODS: The records of 81 adult blunt trauma patients with Glasgow Coma Scale scores < or = 8 and transport times < 2 hours to a Level I trauma center were retrospectively reviewed searching for the following 11 secondary brain injury factors (SBIFs) in the first 24 hours postinjury: hypotension, hypoxia, hypercapnia, hypocapnia, hypothermia, hyperthermia, metabolic acidosis, seizures, coagulopathy, hyperglycemia, and intracranial hypertension. We recorded the worst SBIF during six time periods: hours 1, 2, 3, 4, 5 to 14, and 16 to 24. Occurrence of each SBIF was then correlated with outcome. RESULTS: Hypocapnia, hypotension, and acidosis occurred more frequently than other SBIFs (60-80%). Hypotension, hyperglycemia, and hypothermia were associated with increased mortality rate. Patients with episodes of hypocapnia, acidosis, and hypoxia had significantly longer intensive care unit length of stay (LOS). These three SBIFs and hyperglycemia related to longer hospital LOS as well. Hypotension and acidosis were associated with discharge to a rehabilitation facility rather than home. Finally, multivariate regression analysis revealed that hypotension, hypothermia, and Abbreviated Injury Scale score of the head were independently related to mortality, whereas other SBIFs, age, Injury Severity Score, and Glasgow Coma Scale score were not. Metabolic acidosis and hypoxia were related to longer intensive care unit and hospital LOS. CONCLUSION: Our early management of head injured patients stresses avoidance and correction of SBIFs at all costs. Nonetheless, SBIFs occur frequently in the first 24 hours after traumatic brain injury. Six of the 11 factors studied are associated with significantly worse outcomes. Hypotension and hypothermia are independently related to mortality. Because these SBIFs are potentially preventable, protocols could be developed to decrease their frequency. PMID- 12579058 TI - Influence of low tidal volumes on gas exchange in acute respiratory distress syndrome and the role of recruitment maneuvers. AB - BACKGROUND Use of a low tidal volume (V(T)) strategy in the treatment of acute respiratory distress syndrome can lead to a decrease in oxygenation. This study evaluated the safety and efficacy of a recruitment maneuver (RM) in this setting. METHODS: Twelve patients with acute respiratory distress syndrome were studied within 48 hours of diagnosis. Baseline gas exchange, hemodynamics, and respiratory mechanics were determined and patients were placed on a V(T) of 6 mL/kg. Measurements were repeated and an RM of 30 cm H20 for 40 seconds was performed. Measurements were repeated at 30 minutes and 2 hours post-RM. RESULTS: Decreasing V(T) resulted in a decrease in arterial oxygenation (from 91 +/- 9 mm Hg to 75 +/- 9 mm Hg, p < 0.01), an increase in shunt (from 19 +/- 3.7% to 23 +/- 5%, p < 0.01), and a decrease in lung compliance (from 37 mL/cm H2O to 33 mL/cm H2O, < 0.05). At 30 minutes post-RM, oxygenation improved to 99 +/- 16 mm Hg, shunt decreased to 17 +/- 3%, and lung compliance improved to 39 mL/cm H2O (p < 0.05). Two hours later, oxygenation fell (86 +/- 12 mm Hg), shunt increased (20 +/- 3%), and compliance diminished (36 mL/cm H2O). There were no hemodynamic or barotraumatic complications. CONCLUSION: An RM transiently improves gas exchange during low V(T)ventilation. RMs are well tolerated and no hemodynamic consequences were seen. PMID- 12579059 TI - A rationale for lung recruitment in acute respiratory distress syndrome. PMID- 12579060 TI - Detection of intra-abdominal injury using diagnostic peritoneal lavage after shotgun wound to the abdomen. AB - BACKGROUND: The utility of diagnostic peritoneal lavage (DPL) as a diagnostic tool specifically for shotgun wound to the abdomen (SGWA) is unknown. This prospective study was undertaken to determine the sensitivity, specificity, and accuracy of DPL for the detection of intra-abdominal injuries following SGWA. METHODS: DPL was performed on all patients sustaining SGWA who lacked a clear indication for laparotomy. Patients exceeding 10,000 red blood cells (RBC)/mm were taken for exploratory laparotomy. A prospective database was kept with information on wound location, DPL result, findings upon laparotomy and outcome. RESULTS: Thirty-two DPLs were performed at our urban Level I trauma center for SGWA. Of these, 8 patients had a positive DPL. Upon laparotomy, 7 patients were found to have intra-abdominal injuries, 6 of which required surgical intervention. One patient had no peritoneal penetration or intra-abdominal injury. Of the 24 patients that had a negative DPL, 1 subsequently developed indications for laparotomy and was found to have operative injuries. For predicting intra-abdominal injuries DPL has a sensitivity, specificity and accuracy of 87.5%, 95.8% and 93.8%, respectively. CONCLUSION: For patients presenting with SGWA who do not present with indications for immediate laparotomy, DPL is a reliable indicator of intra-abdominal injury and need for operative intervention. PMID- 12579061 TI - Initial management of massive oral bleeding after midfacial fracture. AB - BACKGROUND: This article reviews initial outcomes of the treatment of massive oral bleeding after midfacial fracture. METHODS: Massive bleeding was defined according to the criteria of Buchanan and Holtmann. The incidence of patients who met these criteria, hematocrit, the type of fracture, time from injury to initial management, source of bleeding, duration of management, and treatment strategy were recorded. RESULTS: We identified massive bleeding in the maxillofacial region in 5 of 521 patients (0.96%). All patients demonstrated profuse bleeding from the nose, and no active source of bleeding was evident in the oral cavity. Bleeding was controlled by nasal packing and temporary reduction in all patients, none of whom required artery ligation or embolization. CONCLUSION: Massive oral bleeding associated with midfacial fractures is frequently derived from the nasal cavity and associated structures. Although the nose may be the cause of the bleeding, the sinuses, skull base, and nasopharynx may also have active bleeding that has cleared through the nasal cavity and nasopharynx into the oral cavity. Control of this massive nasal bleeding during the early stage can therefore improve morbidity associated with severe exsanguination. PMID- 12579062 TI - Adverse drug events in trauma patients. AB - BACKGROUND: Adverse drug events (ADEs) are noxious and unintended results of drug therapy. ADEs have been shown to be a risk to hospitalized patients. The purpose of this study was to determine the rate and nature of ADEs in trauma patients and to characterize the population at risk. METHODS: An electronic medical record, a hospital wide computerized surveillance program, and a clinical pharmacist prospectively investigated ADEs in 4,320 trauma patients from 1996 through 1999. RESULTS: The rate of ADEs in trauma patients (98/4320, 2.3%) was twice that of non-trauma hospital patients (1,111/96,218, 1.2%, p < 0.001). Traumatized females had ADEs 1.5 times more often than traumatized males (2.7% versus 1.8%, p = 0.052). The medication class most often associated with ADEs was analgesics with 54% involving morphine and 20% involving meperidine. The most common ADEs were nausea, vomiting, and itching. Only one ADE was directly attributed to a medical error. CONCLUSIONS: Trauma patients are at double the risk for ADEs. Analgesics are particularly associated with ADEs and use should be carefully monitored. PMID- 12579063 TI - System compliance with out-of-hospital trauma triage criteria. AB - BACKGROUND: Emergency Medical Services (EMS) providers are the initial link to a trauma care system. Previous studies have demonstrated poor compliance with trauma triage by EMS personnel. We sought to determine the proportion of adult EMS cases within a large state meeting Trauma Triage Criteria (TTC) who are ultimately cared for in trauma centers. METHODS: Merged EMS and hospital discharge records for 1996 were examined. All adult acute trauma cases were included. Single-system burns and late effects of injury were excluded. RESULTS: Nine thousand one hundred seventy-four adult cases had at least one TTC, and 60.1% of these patients were transported to a non-trauma center (NTC) and 74.6% of cases with an Injury Severity Score > 15 and one TTC were taken to trauma centers. Analyzing two large urban counties, 58.2% and 27.0% of all TTC cases were still taken to NTC hospitals. CONCLUSION: A significant proportion of seriously injured patients meeting TTC were transported by EMS personnel to NTCs. PMID- 12579064 TI - Victims of domestic violence on the trauma service: unrecognized and underreported. AB - BACKGROUND: Domestic violence (DV) has received increased recognition as a significant mechanism of injury. To improve awareness about DV at our institution, an educational program was presented to the departments of surgery and emergency medicine. Pre and posttests were given and improvement in knowledge was demonstrated. In addition, a screening question for DV was added to the trauma history and physical (H & P) form. This study was done to determine the long-term efficacy of these efforts in increasing recognition of DV and referral to social services in patients admitted to the trauma service. Recognition of DV and appropriate referral should be increased after education and change in H & P form. METHODS: All patients admitted to the trauma service at a Level I trauma center over a 10 month period with the mechanism of injury "assault" were reviewed. DV was determined to be present, likely, unknown, or absent based on information from the prehospital report and medical records. The DV screen question was reviewed for use and accuracy. RESULTS: During the study period, 1,550 patients were admitted to the trauma service, with assault listed as the mechanism of injury for 217 (14%). DV was confirmed or likely in 27 patients (12.4% of the assaults). Of patients with confirmed or likely DV, only 7 received appropriate referrals, with 2 generated by the nursing staff. Of the confirmed and likely DV patients, 17 (63%) were sent home without investigation of safety and only 21% of all assault victims had any social services evaluation (usually to investigate funding or placement). The DV screen was used in only 12 patients. Reasons given for failure to complete the DV screen on the H & P included examiner discomfort in asking the question, and an environment judged to be inappropriate (resuscitation area in the emergency department). CONCLUSION: DV is unrecognized and underreported. Efforts to improve recognition and reporting of DV events need to be ongoing. Screening for DV is not effectively done as part of the initial evaluation. Assessment for DV may be more appropriate as part of the tertiary survey. PMID- 12579065 TI - Arthroscopically assisted osteosynthesis for tibial plateau fractures. AB - BACKGROUND: Options for management of tibial plateau fracture are numerous, and the outcome depends on several factors. With the inherent advantages of arthroscopy, the results of tibial plateau fracture have been greatly improved. METHODS: Thirty-one patients were treated with arthroscopically assisted reduction for tibial plateau fracture, with an average follow-up period of 3 years. Evaluations were performed with the Hospital for Special Surgery knee score clinically, and with the Ahlback scale for osteoarthritis radiographically. RESULTS: Eighty percent of cases were Schatzker type II, III, or IV, and over half (52%) of the patients had concomitant intra-articular lesions. Twenty-nine patients (93.5%) had satisfactory results according to the Hospital for Special Surgery knee score, and only one patient developed degenerative arthritis during the period of follow-up. CONCLUSION: The arthroscopically assisted reduction of tibial plateau fracture is a safe and promising procedure. The results are comparable to traditional methods of open reduction. PMID- 12579066 TI - Isolated adrenal gland injury from penetrating trauma. PMID- 12579067 TI - Possibilities, problems, and pitfalls in surgical management of severe liver trauma illustrated by the report of an exemplary case. PMID- 12579068 TI - Iatrogenic tracheal rupture: a novel approach to repair and a review of the literature. PMID- 12579069 TI - Fracture-dislocation of the glenohumeral joint in a 2-year-old child: case report. PMID- 12579070 TI - Locked pubic symphysis with ipsilateral fracture neck of a femur. PMID- 12579071 TI - The orthoplastic approach for management of the severely traumatized foot and ankle. PMID- 12579072 TI - Practice management guidelines for geriatric trauma: the EAST Practice Management Guidelines Work Group. PMID- 12579073 TI - Shotgun pellet embolus to the middle cerebral artery: case report. PMID- 12579074 TI - Retained thoracic missile: should the bullet be removed? PMID- 12579075 TI - Trauma in pregnancy: anesthetic management of the parturient with multiple gun shot wounds to the gravid uterus and fetal injury. PMID- 12579076 TI - [Allergy risk of transgenic foods, methods for prevention]. PMID- 12579077 TI - [The optimal dose of 5% imiquimod for superficial basal cell carcinomas] PMID- 12579078 TI - [Assessment of the injunction regarding genetic fingerprinting of sex offense perpetrators]. AB - OBJECTIVE: To assess the problems eventually created by fingerprinting for genetic identification of suspected sexual aggressors. METHOD: An opinion poll was conducted among 13 practitioners (8 forensic experts and 5 biologists) using a list of 20 questions. RESULTS: Ten practitioners agreed that there were ethical problems in fingerprinting, and 6 of the 13 physicians surveyed claimed that fingerprinting was distressful for the suspect. COMMENTS: The ethical problems most frequently encountered were the presence of a third party during fingerprinting, the practitioners' lack of information at to the test's future use, the suspect's refusal to undergo such a test and the suspect's lack of information on the subsequent computerized treatment of the fingerprint, to enhance a national genetic data base for example, and the length of time during which the fingerprint is stored. PMID- 12579079 TI - [Prehospital noninvasive ventilation. Study of importance and feasibility (7 cases)]. AB - INTRODUCTION: Non-invasive ventilation can make it possible to avoid endotracheal intubation in patients with acute respiratory failure. Its use in mobile intensive care unit requires assessment of its feasibility. OBJECTIVES: To evaluate the feasibility of noninvasive ventilation in mobile intensive care unit among patients presenting an acute respiratory failure of varied causes. To evaluate the specific problems and to propose adapted answers. METHODS: Seven patients presenting with acute respiratory failure were included in a prospective study conducted with the SAMU (mobile intensive care unit) in Bordeaux between July 01 and November 01, 2000. Non-invasive ventilation was applied, in the absence of contraindications, using a standard facial mask to help inspiration and with added positive expiration pressure. Efficacy was assessed on clinical and biological criteria. RESULTS: Non-invasive ventilation improved the respiratory state in most patients and avoided endotracheal intubation in 6 patients. DISCUSSION: This study made it possible to advance criteria that we feel are essential with the implementation of this method of respiratory assistance, and provided solutions to the various problems encountered (equipment, training, protocols and follow-up of patients). CONCLUSION: Despite the limits of our study we were able to demonstrate the feasibility of non invasive ventilation in prehospital setting. Future studies should be conducted on larger cohorts in order to specify the interest of non-invasive ventilation in such conditions. PMID- 12579081 TI - [Apropos of a rare tumor of the pleura: benign schwannoma]. PMID- 12579080 TI - [Isolated adenopathy in leishmaniasis due to Leishmania infantum]. AB - INTRODUCTION: Leishmaniasis is rarely manifested by isolated lymphadenitis in the mediterranean area, only 13 cases have been reported. OBSERVATION: An isolated cervical lymphadenitis was observed in a 37 year-old man living in the north of Tunisia. The fine needle aspiration of the enlarged lymph node found Leishmania in both direct examination and culture. Isoenzymatic typing identified Leishmania infantum MON-1. The evolution was slowly favourable without treatment. DISCUSSION: Despite the rare number of cases described, looking for leishmaniasis should be systematic when confronted with lymphadenitis in endemic areas. The therapeutic decision depends on the immunological status of the patients and the possibility of regular medical controls. PMID- 12579082 TI - [Hepatic colic revealing vesicular metastasis of melanoma]. PMID- 12579083 TI - [Bilateral conjunctival involvement revealing non-Hodgkin's lymphoma]. PMID- 12579084 TI - [Cutaneous tumors in organ transplant recipients]. AB - THE CONTEXT: With the improvement in transplant in surgery, organ storage techniques and immunosuppressive treatments, the number of organ transplant recipients is regularly increasing. To preserve the activity of the graft, immunosuppressive treatment is required for life in most cases, leading to prolonged immunodeficiency. These patients exhibit an increased risk of cancer, notably of tumors associated with viruses such as carcinomas, Kaposi's sarcoma and lymphomas. THE DIFFERENT CANCERS CONCERNED: Cutaneous cancers, notably squamous cell carcinomas, are the predominant cancers encountered in transplant recipients and affect around half of the patients within 20 years following the transplant. Squamous cell carcinomas tend to be multiple, may be life threatening, and appear to predict the onset of other cancers. Exposure to UV, immunosuppression, the relatively old age at the time of the transplant, light skinned phototype, genetic factors and papillomavirus infections are all factors of risk. Other photo-induced tumors, such as melanoma, some sarcomas or Merkel cell carcinomas also appear increased and of severe prognosis. MEASURES OF PREVENTION: The prevalence of these tumors could be reduced by educating the patients on strict sun-protection measures following the transplant. The factors enhancing Kaposi's sarcoma associated with the HHV8 virus remain to be defined. The possibility of transmission of the infection through the graft must lead to the systematic search for this virus in the donors. PMID- 12579085 TI - [Blood doping and cardiovascular consequences]. AB - THE REASONS FOR BLOOD DOPING: The aim of doping the hematological functions is to increase the quantity of oxygen supplied to the muscles in activity. THE DOPING METHODS: Self blood transfusion was one of the first methods used. The incriminated substances are blood substitutes, non-specific and specific hematopoietic growth factors such as erythropoietin and granulocyte macrophage colony stimulating factor, and other substances such as perfluorocarbon and RSR13. DELETERIOUS CONSEQUENCES: The major risk is that of a hyperviscosity syndrome, responsible for thrombosis and decreased cardiac output. IN PRACTICE: All physicians are concerned, since they may be confronted with the complications of doping in their daily practice. They should also know the list of prohibited substances, in order to avoid any errors in prescription. PMID- 12579086 TI - [Telithromycin, first ketolide to diversify the management of community-acquired respiratory tract infections. Importance in emergency care]. PMID- 12579087 TI - [Hormone replacement therapy in menopause and cardiovascular diseases]. PMID- 12579089 TI - Multicenter bridge to transplantation with the HeartMate assist device: evaluation from another perspective. PMID- 12579090 TI - Chronic and adjustable pulmonary artery banding. AB - OBJECTIVE: Banding of the pulmonary artery might be required to prevent pulmonary vascular damage in patients with increased pulmonary artery flow and to retrain the left ventricle in preparation for an arterial switch operation in patients with congenitally corrected transposition of the great arteries. Readjustment of the pulmonary artery band might be required in the postoperative period. In this study we aimed to test the feasibility of a novel device for bidirectionally adjustable pulmonary artery constriction. METHODS: A hydraulic main pulmonary artery occluder was implanted in lambs and gradually inflated to create right ventricular pressure overload at a systemic (aortic) level. During the following period (up to 12 weeks), this pressure overload was monitored by measuring aortic and right ventricular pressures by means of implanted subcutaneous reservoirs. If required to maintain the right ventricular pressure overload at a systemic level in the growing animals, the occluder was deflated through a third subcutaneous reservoir. RESULTS: After the banding period (average of 64 +/- 8 days), the main pulmonary artery cuff could still be adjusted, and the animals showed no clinical signs of heart failure. Histologic analysis of the pulmonary artery showed extensive fibrosis, a giant cell response around the device, and small areas of tissue necrosis; complete transmural necrosis was not detected. CONCLUSIONS: This device allows adjustment of the pulmonary artery cuff in a precise manner over a prolonged period of time without surgical reintervention. Potentially, the device might have applications for clinical use in children with congenital heart disease. PMID- 12579091 TI - Surgical management of scimitar syndrome: an alternative approach. AB - OBJECTIVE: The scimitar syndrome is a congenital anomaly that consists in part of total or partial anomalous venous drainage of the right lung to the inferior vena cava. Surgical approaches to the scimitar syndrome have varied according to the anatomic and pathologic features presented in each case. The aim of this study was to present an alternative approach to the surgical correction of scimitar syndrome. METHODS: Nine patients with the scimitar syndrome were operated on between 1990 and 2000. They comprised 1 male and 8 female patients (mean age 11.5 +/- 17.6 years). All patients had symptoms, with recurrent pneumonia or respiratory tract infections and pulmonary/systemic flow ratios greater than 1.5:1.0. None of the patients had pulmonary hypertension or an atrial septal defect. All patients underwent repair of the anomalous scimitar vein by direct reimplantation into the left atrium without cardiopulmonary bypass. Two patients underwent concomitant resection of a right lower lobe sequestration. Follow-up was complete in all cases. RESULTS: There were no operative or late deaths, and no patients have required reoperation. At the time of follow-up (mean 55 +/- 46 months), echocardiography demonstrated a patent anastomosis in all patients without any evidence of restenosis. CONCLUSION: This clinical experience indicates that an alternative surgical approach to scimitar syndrome is direct anastomosis of the scimitar vein to the posterior aspect of the left atrium using a right thoracotomy without cardiopulmonary bypass. This procedure is safe and effective and obviates the need for long intra-atrial baffles and the use of the extracorporeal circuit. PMID- 12579092 TI - Vascular endothelial growth factor and basic fibroblast growth factor expression in esophageal adenocarcinoma and Barrett esophagus. AB - OBJECTIVE: This study was undertaken to investigate the role of the angiogenic factors vascular endothelial growth factor and basic fibroblast growth factor in the development and progression of Barrett esophagus and adenocarcinomas of the esophagus and gastroesophageal junction. METHODS: Vascular endothelial growth factor and basic fibroblast growth factor messenger RNA expression levels, relative to the control gene encoding beta-actin, were measured by using a quantitative reverse transcription-polymerase chain reaction method (ABI 7700 Sequence Detector system) in specimens of Barrett intestinal metaplasia (n = 16), dysplasia (n = 11), adenocarcinoma (n = l 5), and matching normal squamous esophageal tissues (n = 35). Vascular endothelial growth factor and basic fibroblast growth factor protein expression and CD31(+) microvessel density were assessed by means of immunohistochemistry in 25 tissue sections that included representative areas for each of these Barrett stages. RESULTS: Expression levels were significantly increased in adenocarcinoma compared with in either normal squamous mucosa (P <.0001 for both genes) or intestinal metaplasia (vascular endothelial growth factor, P =.002; basic fibroblast growth factor, P <.0001). Vascular endothelial growth factor levels were also significantly higher in cancer tissues compared with dysplasia tissues (P =.024, Mann-Whitney U test). Basic fibroblast growth factor expression was also significantly increased in Barrett dysplastic mucosa compared with in intestinal metaplasia or normal esophageal mucosa. Microvessel density was generally higher in adenocarcinoma compared with in preneoplastic Barrett tissues. The pattern of vascular endothelial growth factor and basic fibroblast growth factor protein expression was similar to the messenger RNA expression pattern, with the exception that mucin-containing goblet cells stained intensely for vascular endothelial growth factor and only weak vascular endothelial growth factor staining was present in some adenocarcinomas. CONCLUSIONS: Vascular endothelial growth factor and basic fibroblast growth factor messenger RNA expression levels are significantly upregulated in esophageal and gastroesophageal junction adenocarcinomas, suggesting a role for these angiogenic factors in the development of these cancers. Vascular endothelial growth factor and basic fibroblast growth factor messenger RNA expression levels are also increased in some Barrett esophagus tissues, with this increase occurring at an earlier stage for basic fibroblast growth factor than for vascular endothelial growth factor. Basic fibroblast growth factor protein expression pattern is similar to the messenger RNA expression pattern, but unlike the messenger RNA findings, vascular endothelial growth factor protein expression is strongest in goblet cells. PMID- 12579093 TI - A randomized trial comparing induction chemotherapy followed by surgery with surgery alone for patients with stage IIIA N2 non-small cell lung cancer (JCOG 9209). AB - OBJECTIVE: We performed a prospective randomized trial in patients with potentially resectable stage IIIA N2 non-small cell lung cancer to confirm the efficacy of induction chemotherapy before surgical resection. METHODS: Patients with stage IIIA N2 non-small cell lung cancer, all with histologically or cytologically confirmed metastases to the ipsilateral mediastinal lymph nodes, were randomly assigned to receive either three cycles of induction chemotherapy (cisplatin at 80 mg/m(2) on 1 day and vindesine at 3 mg/m(2) on 2 days) followed by surgery or surgery alone. RESULTS: This trial was prematurely terminated because the accrual rate was too slow, which lowered the study's statistical power considerably. From June 1993 through April 1998, a total of 62 patients were enrolled, and 31 patients were assigned to each treatment group. The objective clinical response rate of induction chemotherapy was 28%. Complete resection was achieved in 20 patients in the induction chemotherapy group (65%) and 24 in the surgery alone group (77%). Median follow-up was 6.2 years. Median overall survivals were 17 months for the induction group and 16 months for the surgery alone group. The estimated 1-, 3-, and 5-year survivals, respectively, were 68% (95% confidence interval 51%-85%), 23% (95% confidence interval 8%-38%), and 10% (95% confidence interval 0%-20%) for the induction chemotherapy group and 65% (95% confidence interval 48%-82%), 26% (95% confidence interval 11%-41%), and 22% (95% confidence interval 7%-37%) for the surgery alone group. There was no statistically significant difference in survival between the groups (P =.5274). Treatment-related death was not observed in either group. CONCLUSION: This randomized trial to compare induction chemotherapy (cisplatin and vindesine) followed by surgery with surgery alone for patients with stage IIIA N2 non-small cell lung cancer did not demonstrate a survival difference between the groups, although this may have been because the statistical power was limited. PMID- 12579094 TI - The role of proinflammatory cytokines in lung ischemia-reperfusion injury. AB - OBJECTIVE: Proinflammatory cytokines are known to play roles in ischemia reperfusion injury of the heart, kidney, small bowel, skin, and liver. Little is known about their roles in ischemia-reperfusion injury of the lung. This study was undertaken to define the role of 2 proinflammatory cytokines, tumor necrosis factor alpha and interleukin 1beta, in ischemia-reperfusion injury of the lung. METHODS: Left lungs of male rats were rendered ischemic for 90 minutes and reperfused for up to 4 hours. Treated animals received anti-tumor necrosis factor alpha or anti-interleukin 1beta antibody before reperfusion. Increased vascular permeability in the lung was measured by using iodine 125-labeled bovine serum albumin. Neutrophil sequestration in the lung parenchyma was determined on the basis of activity. Bronchoalveolar lavage was performed to measure cell counts. Separate tissue samples were processed for histology, cytokine protein, and messenger RNA content by using Western blotting and the ribonuclease protection assay. RESULTS: Animals receiving anti-tumor necrosis factor alpha and anti interleukin 1beta demonstrated reduced injury compared with that seen in positive control animals (vascular permeability of 48.7% and 29.4% lower, respectively; P <.001). Vascular injury was reduced by 71% when antibodies to tumor necrosis factor alpha and interleukin 1beta were administered together. Lung neutrophil accumulation was markedly reduced among animals receiving anti-tumor necrosis factor alpha and anti-interleukin 1beta (myeloperoxidase content of 30.9% and 38.5% lower, respectively; P <.04) and combination blockade afforded even greater protection (52.4% decrease, P <.01). Bronchoalveolar lavage leukocyte content was also reduced by treatment with anti-tumor necrosis factor alpha, anti-interleukin 1beta, and combination treatment. Reductions in permeability, myeloperoxidase, and bronchoalveolar lavage leukocyte content also resulted in a decrease in a histologic injury. Finally, anti-tumor necrosis factor alpha and anti-interleukin 1beta treatment resulted in decreased messenger RNA expression for a number of early response and regulatory cytokines. CONCLUSION: Tumor necrosis factor alpha and interleukin 1beta help regulate the development of lung ischemia-reperfusion injury. They appear to promote injury by altering expression of proinflammatory and anti-inflammatory cytokines and influencing tissue neutrophil recruitment. PMID- 12579095 TI - Endoscopic mitral valve repair: feasible, reproducible, and durable. AB - OBJECTIVE: We sought to document the feasibility, safety, and effectiveness of performing mitral valve repair using a totally endoscopic approach. METHODS: Between February 1997 and October 1, 2001, 187 patients underwent totally endoscopic mitral valve repair at our institution. The mean age was 60.7 +/- 13.1 years, and 62% were male. Median preoperative functional class and degree of mitral regurgitation were II and 4, respectively. Data collection included an institutional protocol assessing procedure-related pain, cosmesis, and functional recovery. Statistical analysis included Kaplan-Meier and Cox regression methods. Mean follow-up was 19 +/- 15.2 months and was 100% complete. RESULTS: Associated atrial procedures were performed in 9.1% (n = 17) of the patients. Two patients required intraoperative conversion to sternotomy. Thoracoscopic re-evaluation for suspected bleeding (n = 19) was part of our aggressive postoperative management. One patient required sternotomy for control of bleeding. Hospital mortality included 1 (0.5%) patient and was not technology related. There were 1 early and 6 late reoperations, 4 of which were due to endocarditis. No risk factors for repair failure could be detected. Freedom from mitral valve reoperation at 4 years was 93.3% +/- 2.6%. The median degree of mitral regurgitation at follow-up was 0. Ninety-three percent of the patients were highly satisfied with either no or mild postoperative pain, and 98.4% believed they had an aesthetically pleasing scar. CONCLUSIONS: Totally endoscopic mitral valve repair can be done safely with excellent results and a high degree of patient satisfaction. It is now our exclusive approach for isolated atrioventricular valve disease. PMID- 12579096 TI - Acute volume reduction with aortic valve replacement immediately improves ventricular mechanics in patients with aortic regurgitation. AB - OBJECTIVES: Few data have been available regarding the immediate response in ventricular mechanics to acute volume reduction caused by aortic valve replacement for aortic regurgitation. METHODS: We studied 9 patients in the operating room immediately before and after the institution of cardiopulmonary bypass. Left ventricular pressure and cross-sectional area (a surrogate of left ventricular volume) were measured with a catheter-tip manometer and a transesophageal echocardiographic system equipped with automated border-detection technology. Left ventricular pressure-area loops were constructed, and the caval occlusion method was used to obtain the slope of the end-systolic pressure-area relationship and the end-systolic area associated with 100 mm Hg. From the steady state beats, stroke area was obtained by subtracting the minimum area from the maximum area. Effective arterial elastance, a measure of ventricular afterload, was calculated from end-systolic pressure, and stroke area as follows: effective arterial elastance equals end-systolic pressure divided by stroke area. RESULTS: Reductions in maximum area (21.0 +/- 8.5 to 16.0 +/- 6.8 cm(2) [SD])and minimum area (15.3 +/- 8.4 to 12.0 +/- 6.1 m(2)) shifted the baseline pressure-area loops to the left. The slope of the end-systolic pressure-area relationship (11.6 +/- 4.8 to 16.0 +/- 7.5 mm Hg/cm(2)) and afterload (effective arterial elastance, 17.9 +/- 11.6 to 26.3 +/- 16.4 mm Hg/cm(2)) were increased, and the end-systolic area associated with 100 mm Hg was reduced (18.3 +/- 10.0 to 13.7 +/- 5.8 cm(2)). CONCLUSION: Correction of volume overload reduced preload (minimum area), shifted the end-systolic pressure-area relationship to the left (decreased end-systolic area), and improved ventricular contractility (increased slope of the end systolic pressure-area relationship). The result indicated that acute volume reduction favorably influenced ventricular mechanical parameters immediately after the operation. PMID- 12579097 TI - "Our complication rates are lower than theirs": statistical critique of heart valve comparisons. AB - OBJECTIVES: From the widely differing complication rates published for every heart valve, is it possible to determine a true rate for each valve and to compare the rates of two different valves? We investigated this question for the two most popular bileaflet valves. METHODS: Aortic valve data were abstracted from 14 St Jude Medical (St Jude Medical Inc, Minneapolis, Minn; 33,125 patient y) and 11 Carbomedics (Sulzer Carbomedics Inc, Austin, Tex; 19,141 patient-y) series, and mitral valve data were abstracted from 11 St Jude Medical (21,553 patient-y) and 8 Carbomedics (8368 patient-y) series. Regression analysis was used to accommodate heterogeneity among rates with the same valve model, to estimate hazard ratios and 95% confidence intervals for the valve model effect, and to incorporate other series-level risk factors. RESULTS: Most of the complication rates with both valve models exhibited significant heterogeneity. For thromboembolism and bleeding, the relative risks for valve model were not significantly different from unity. Valve thrombosis rates exhibited less heterogeneity: the Carbomedics valve had a lower rate in the aortic position (hazard ratio 0.2, 95% confidence interval 0.1-0.6) and a higher rate in the mitral position (hazard ratio 1.9, 95% confidence interval 1.0-3.8). CONCLUSIONS: Unlike simple weighted averages of valve complication rates, regression methods can incorporate heterogeneity related to center effects and allow for inclusion of other risk factors. Thromboembolism and bleeding rates were not significantly different with St Jude Medical and Carbomedics valves. Valve thrombosis appeared to differ between the two valves, but the absolute differences in rates were small. Because of the variability among rates with the same valve, statistical comparisons must be interpreted cautiously. PMID- 12579098 TI - Vacuum-assisted closure as a treatment modality for infections after cardiac surgery. AB - OBJECTIVE: Wound infections after cardiac surgery carry high morbidity and mortality. A plethora of management strategies have been used to treat such infections. We assessed the impact of vacuum-assisted closure on the management of sternal wound infections in terms of wound healing, duration of vacuum assisted closure, and cost of treatment. METHODS: Between November 1998 and June 2001, a total of 27 mediastinal infections were managed with vacuum-assisted closure. Group A (n = 14) had vacuum-assisted closure as the final treatment modality, whereas in group B (n = 13) vacuum-assisted closure was followed by either a myocutaneous flap (n = 8) or primary (n = 5) wound closure. The choice of additional treatment modality was based on wound size. RESULTS: In group A, 4 patients died and a satisfactorily healed scar was achieved in 64% of cases. Median durations of vacuum-assisted closure and hospital stay in group A were 13.5 days (interquartile range 8.8-32.2 days) and 20 days (interquartile range 16.7-25.2 days), respectively. Mortality was 7.7% in group B, with a treatment failure rate of 15%. Median duration of vacuum-assisted closure in group B was 8 days (interquartile range 5.5-18 days), and median hospital stay was 29 days (interquartile range 25.8-38.2 days). During the year before institution of vacuum-assisted closure, poststernotomy infection (n = 13) was managed with rewiring and closed irrigation system. Treatment during this year failed in 30.7% of cases (n = 4/13), and mortality was also 30.7%. The total cost (hospitalization and treatment) per patient for vacuum-assisted closure was 16,400 dollars, compared with 20,000 dollars for the closed irrigation system treatment. CONCLUSION: Vacuum-assisted closure, used alone or before other surgical treatment strategies, is an acceptable treatment modality for infections in cardiac surgery with reasonable morbidity, mortality, and cost. PMID- 12579099 TI - Mechanisms of Gal(alpha)1-3Gal(beta)1-4GlcNAc-R (alphaGal) expression on porcine valve endothelial cells. AB - OBJECTIVE: We have previously reported that porcine valve endothelium does not express immunodetectable levels of the carbohydrate Gal(alpha)1-3Galbeta1-4GlcNAc R (known as alphaGal), suggesting that fresh porcine valve may be immunoprivileged. In this study, we further investigated the mechanisms of alphaGal expression on porcine valve endothelial cells. METHODS: Primary cultures of porcine valvular endothelial cells were established and compared with porcine aortic endothelial cells and human vein endothelial cells. Immunoblotting, reverse transcriptase-polymerase chain reaction, and flow cytometry were used to compare the expression of alphaGal at both the protein and messenger RNA levels. RESULTS: Porcine valvular endothelial cells grew rapidly on a gelatin substrate. Similar to our previous in vivo results, valve endothelial cells expressed alphaGal much less intensely than did aortic endothelial cells. Porcine aortic endothelial cells expressed an isolectin B4 (isolectin B4 lectin Bandeiraea simplicifolia) immunodetectable band at 135 kd that was not visible on porcine valve endothelial cells or on human vein endothelial cells. Reverse transcriptase polymerase chain reaction documented three transcripts of the alphaGal gene that were identically expressed on porcine valve and aortic endothelial cells. Furthermore, flow cytometry showed an almost identical surface profile between porcine aortic and valve endothelial cells, in contrast with human vein endothelial cells. CONCLUSIONS: Cultures of primary valve endothelial cells were established and exhibited similar phenotypic patterns in vitro to those we have previously documented in vivo. RNA and flow cytometric analyses documented no difference between the RNA expression and surface protein profile for alphaGal, although whole-cell extracts demonstrated an immunodetectable band on Western blotting that was present on aortic endothelial cells but not on valve endothelial cells. These findings clarify the mechanism of expression of alpha1,3galactosyltransferase gene expression in valve endothelial cells, suggesting that delayed rejection of fresh porcine cardiac valves may occur. PMID- 12579100 TI - Tachycardia-induced cardiomyopathy in the ovine heart: mitral annular dynamic three-dimensional geometry. AB - BACKGROUND: Ring annuloplasty has been used to correct annular dilatation and mitral regurgitation in dilated cardiomyopathy, but little is known about the dynamic precise 3-dimensional geometry of the mitral annulus in this condition. METHODS: Nine sheep had radiopaque markers sewn to the mitral annulus, creating 8 distinct segments beginning at the posterior commissure (segments 1-4, septal mitral annulus; segments 5-8, lateral mitral annulus). Biplane videofluoroscopy and transesophageal echocardiography were performed before and after rapid pacing (180-230 min(-1) for 15 +/- 6 days) sufficient to develop tachycardia-induced cardiomyopathy and mitral regurgitation. Mitral annular segment contraction was defined as the percentage difference between maximum and minimum lengths. Mitral annular area and mitral annular septal-lateral and commissure-commissure diameters and 3-dimensional shape were determined from marker coordinates. RESULTS: With tachycardia-induced cardiomyopathy, end-diastolic mitral annular area, septal-lateral diameter, and commissure-commissure diameter increased by 36% +/- 14%, 25% +/- 12%, and 9% +/- 5%, respectively (P <.01), whereas mitral regurgitation increased from 0.3 +/- 0.2 to 2.2 +/- 0.9 (P <.0001). All annular segments dilated at end-diastole with tachycardia-induced cardiomyopathy, except the segment between the midseptal annulus and the left fibrous trigone. Annular segment contraction was significantly decreased with tachycardia-induced cardiomyopathy in the lateral, but not in the septal, regions. Three-dimensional reconstruction of annular shape revealed a saddle shape of the annulus at baseline; this shape was also measured with tachycardia-induced cardiomyopathy, but there was some flattening of the septal annulus. CONCLUSIONS: With tachycardia-induced cardiomyopathy, the mitral annulus dilated substantially, being more in the septal-lateral than in the commissure-commissure dimension. Greater annular segmental dilatation and decreased contraction occurred in the lateral annulus. The saddle shape of the annulus was retained but flattened. PMID- 12579101 TI - Heparin versus danaparoid in off-pump coronary bypass grafting: results of a prospective randomized clinical trial. AB - OBJECTIVE: The incidence of heparin-induced thrombocytopenia is increasing, and the thrombin inhibitor danaparoid could be a useful alternative. The objective of the present study was to compare danaparoid and heparin in patients undergoing off-pump coronary artery bypass grafting. METHODS: In a prospective, randomized, double-blind clinical trial comparing heparin (bolus of 1 mg/kg) with danaparoid (bolus of 40 U/kg), 71 patients underwent off-pump coronary artery bypass grafting with one of the study drugs. The amount of blood lost, the number of homologous blood products transfused, the troponin T levels, and the amount of anti-Xa activity were monitored. RESULTS: Thirty-four patients underwent 2.6 +/- 0.7 bypasses with danaparoid, and 37 patients underwent 2.5 +/- 0.9 grafts with heparin (P =.8). Postoperative blood losses averaged 1394 +/- 1033 mL in patients receiving danaparoid and 1130 +/- 868 mL in patients receiving heparin (P =.2). The number of homologous blood products transfused averaged 3.6 +/- 7 units in patients receiving danaparoid and 1.9 +/- 4.4 units in patients receiving heparin (P =.2). The number of patients requiring homologous blood transfusion was higher in patients receiving danaparoid (18/34 [53%]) than in patients receiving heparin (10/37 [27%], P =.03). Serum anti-Xa activity averaged 1.6 +/- 0.6 U/mL in patients receiving danaparoid and 1.9 +/- 0.8 U/mL in patients receiving heparin 30 minutes after injection of the drugs (P =.1) and 0.3 +/- 0.1 and 0.04 +/- 0.08 U/mL, respectively, 12 hours after coronary artery bypass grafting (P =.001). Troponin serum levels were similar 48 hours after coronary artery bypass grafting (0.5 +/- 0.6 and 0.4 +/- 0.6 microg/L, respectively). CONCLUSION: Although off pump coronary artery bypass grafting with danaparoid versus heparin increases the number of patients exposed to homologous blood transfusion (relative risk, 2; 95% confidence limits, 1-4), off-pump coronary artery bypass grafting with danaparoid is a valuable alternative to heparin in patients with thrombocytopenia requiring surgical intervention. PMID- 12579102 TI - Is vitamin C superior to diltiazem for radial artery vasodilation in patients awaiting coronary artery bypass grafting? AB - OBJECTIVES: We aimed to measure the vasodilating effects of vitamin C on the radial arteries of healthy subjects and to assess whether vitamin C is superior in this regard to diltiazem, a commonly used vasodilator in coronary artery bypass using radial conduits. METHODS: In a case-control study (study 1) oral single-dose vitamin C (2 g) was given to 15 healthy nonsmokers and 15 matched otherwise healthy smokers. In a randomized double-blind study (study 2) oral single-dose vitamin C (2 g, n = 15) and diltiazem (180 mg, n = 15) were compared in preoperative patients with coronary artery disease. We examined the dilation of the radial artery with high-resolution ultrasonography and measurement of the lumen surface and color Doppler images of the nondominant radial artery just before and 2 hours after drug administration. RESULTS: In study 1 both smokers and nonsmokers showed a significant increase in the lumen surface at 2 hours compared with at baseline (P <.001 and P =.013, respectively). The increase was larger in smokers (median, 37.5% vs 14.3%; P =.004). In study 2 both groups showed statistically significant increases in the lumen surface at 2 hours compared with at baseline (P <.001 and P =.008 for vitamin C and diltiazem, respectively). Vitamin C achieved a larger increase than diltiazem (median, 33.3% vs 18.2%; P =.016). In multivariate modeling the increase in lumen surface was independently predicted by use of vitamin C over diltiazem (+21.2%, P =.007), diabetes mellitus (+14.5%, P =.085), increased cholesterol (+26.2%, P =.001), and smoking history (+20.8%, P =.017). CONCLUSIONS: Vitamin C is a potent acute vasodilator in both smokers and nonsmokers and is superior to diltiazem in preoperative coronary patients who need protection from vasospasm of the radial conduit. PMID- 12579104 TI - Intravenous magnesium sulfate prophylaxis for atrial fibrillation after coronary artery bypass surgery. AB - OBJECTIVE: Atrial fibrillation is a rhythm disorder commonly seen early after coronary artery bypass grafting, and it increases morbidity. METHODS: To investigate the effectiveness of magnesium sulfate in the prophylaxis of atrial fibrillation, we conducted a prospective, randomized, placebo-controlled clinical study on 200 consecutive patients in whom we performed elective and initial coronary artery bypass grafting operations. In each group 50% of patients underwent beating-heart operations. In the treatment group 100 patients (76 men and 24 women; mean age, 57.63 +/- 9.68 years) received 24.34 mEq (3 g) of magnesium sulfate in 100 mL of saline solution that was administered over 2 hours (50 mL/h) preoperatively, perioperatively, and at postoperative days 0, 1, 2, and 3. In the control group 100 patients (74 men and 26 women; mean age, 59.96 +/- 9.29 years) received only 100 mL of saline solution according to the same administration schedule as the treatment group. RESULTS: Atrial fibrillation developed in 15 patients from the treatment group and in 16 patients from the control group. The arrhythmia developed after 37.87 +/- 12.76 and 45.26 +/- 15.27 hours in the treatment and control groups, respectively. Although a significant relationship was found between low magnesium sulfate levels and increased incidence of atrial fibrillation (P <.05), when the incidence of postoperative atrial fibrillation is concerned, no significant difference was found between the 2 groups (P >.05). Also, no significant difference was found between operations with cardiopulmonary bypass and beating-heart operations in terms of atrial fibrillation incidence (P >.05). However, atrial fibrillation extended the duration of hospital stay in both groups (P <.05). CONCLUSION: Our findings indicate that magnesium sulfate infusion alone is not sufficient for the prophylaxis of atrial fibrillation. PMID- 12579103 TI - Coagulation and fibrinolytic markers in a two-month follow-up of coronary bypass surgery. AB - OBJECTIVES: The alterations of the coagulation-fibrinolytic profile immediately and up to few days after cardiac surgery have been widely documented. However, less information is available on whether these alterations persist for prolonged periods of time after the operation. In this study we have evaluated the coagulation-fibrinolytic profile of patients who underwent coronary artery surgery with cardiopulmonary bypass during a 2-month follow-up period. METHODS: Twenty-six patients (age range, 50-75 years) were studied. Blood samples were collected before the intervention and at different time points postoperatively up to 2 months after the operation. Measurement of selected coagulation-fibrinolytic variables was carried out in plasma from 16 patients. Evaluation of tissue factor activity determined as procoagulant activity was performed in peripheral blood mononuclear leukocytes obtained from 10 patients. RESULTS: Antigenic levels of clottable fibrinogen, prothrombin fragment F1.2, D-dimer, and thrombin antithrombin complex were significantly increased during the first week after the intervention compared with preoperative values. Prothrombin fragment F1.2 levels returned to normal within 15 days, fibrinogen levels normalized within 30 days, and thrombin-antithrombin complex levels normalized at 45 days, whereas D-dimer values were still significantly higher 60 days postoperatively respective to baseline values. There was a trend toward an increased procoagulant activity from peripheral blood mononuclear leukocytes 4 days after the operation, whereas no changes of factor VII measured either as antigen or in its coagulant and activated forms were recorded throughout the study. CONCLUSIONS: A marked activation of the coagulation-fibrinolytic system occurs after cardiopulmonary bypass and lasts for at least 2 months thereafter. This finding suggests that these alterations might account for the increased thrombotic risk of these patients during the postoperative period. PMID- 12579105 TI - Autosynchronized systolic unloading during left ventricular assist with a centrifugal pump. AB - OBJECTIVES: The purpose of this study was to investigate how the inflow cannulation site of the left ventricular assist system with a centrifugal pump would influence cardiac function on failing heart models. METHODS: In 10 sheep, a left ventricular assist system was instituted by an outflow cannula in the descending aorta, two inflow cannulas in the left atrium and the left ventricle, and connecting those cannulas to a magnetically suspended centrifugal pump. A conductance catheter and a tipped micromanometer for monitoring the pressure volume loop were also inserted into the left ventricle. Myocardial oxygen consumption was directly measured. Heart failure was induced by injection of microspheres into the left main coronary artery. The assist rate was varied from 0% to 100% at each inflow cannulation site. RESULTS: The pump flow with left ventricular cannulation increased during the systolic phase and decreased during the diastolic phase, whereas it was constant with left atrial cannulation. Ejection fraction with left atrial cannulation decreased as the assist rate increased, whereas that with left ventricular cannulation was maintained up to 75% assist. The external work with left atrial cannulation decreased gradually as the assist rate increased, whereas the external work with left ventricular cannulation did not decrease until the assist rate reached 75%. The myocardial oxygen consumption in both cannulations decreased proportionally as the assist rate increased; they were significantly less with left ventricular cannulation at the 100% assist rate than with left atrial cannulation. CONCLUSION: Left ventricular cannulation during left ventricular assistance maintains ejection fraction and effectively reduces oxygen consumption. PMID- 12579106 TI - Long-term improvement of cardiac function in rats after infarction by transplantation of embryonic stem cells. AB - OBJECTIVE: This study was designed to investigate the feasibility of and potential functional improvement with embryonic stem cell transplantation in rats 32 weeks after myocardial infarction. METHODS: Before cell transplantation, cultured embryonic stem cells were transfected with the complementary DNA of green fluorescent protein to identify engrafted cells in myocardium. Myocardial infarction was induced by ligation of the left coronary artery. Either 3 x 10(5) mouse embryonic stem cells or an equivalent volume of cell-free medium was injected into injured myocardium within 20 minutes after induction of myocardial infarction. RESULTS: Embryonic stem cell transplantation significantly increased the survival rate in rats undergoing myocardial infarction during the experimental period of 32 weeks. Hemodynamic and echocardiographic data showed that embryonic stem cell transplantation significantly improved ventricular function relative to the myocardial infarction plus medium control group. Tissue positive for green fluorescent protein was found in the injured myocardium with cell transplantation. The proportion of myocardium positively immunostained by antibodies against alpha-myosin heavy chain and cardiac troponin I was greater in the infarcted area with embryonic stem cell transplantation than in the injured myocardium with medium injection. Single green fluorescent protein-positive cells with a rod shape and clear striations were observed in cardiomyocytes isolated from infarcted hearts with embryonic stem cell transplantation. In addition, the number of blood vessels in injured myocardium was greater in the cell transplanted myocardial infarction group than in the medium-injected myocardial infarction group. CONCLUSIONS: Engrafted embryonic stem cells differentiated into cardiomyocytes in injured myocardium, caused an angiogenetic effect, and subsequently improved cardiac function during the 32-week observation period. PMID- 12579107 TI - Survival and death-promoting events after transient spinal cord ischemia in rabbits: induction of Akt and caspase3 in motor neurons. AB - OBJECTIVE: The mechanism of spinal cord injury has been thought to be related to the vulnerability of spinal motor neuron cells to ischemia. However, the mechanisms of such vulnerability are not fully understood. We previously reported that spinal motor neurons might be lost as a result of programmed cell death and investigated a possible mechanism of neuronal death by means of immunohistochemical analysis for CPP32 (caspase3) and serine-threonine kinase (Akt). METHODS: We used a rabbit spinal cord ischemia model with use of a balloon catheter. The spinal cord was removed at 8 hours or 1, 2, or 7 days after 15 minutes of transient ischemia, and histologic changes were studied with hematoxylin and eosin staining. Western blot analysis for Akt and caspase3, temporal profiles of Akt and caspase3 immunoreactivity, and double-label fluorescence immunocytochemical studies were performed. RESULTS: The majority of motor neurons were preserved until 2 days but were selectively lost at 7 days of reperfusion. Western blot analysis revealed no immunoreactivity for Akt and caspase3 in the sham-operated spinal cords. However, such immunoreactivity became apparent at 8 hours after transient ischemia, decreased at 1 day, and returned to the baseline level at 2 days. A double-label fluorescence immunocytochemical study revealed that both Akt and caspase3 were positive at 8 hours of reperfusion in the same motor neurons, which eventually die. CONCLUSION: These results suggests that transient spinal cord ischemia activates both cell death and survival pathways after ischemia. The activation of Akt protein at the early stage of reperfusion might be one of the factors responsible for the delay in neuronal death after spinal cord ischemia. PMID- 12579108 TI - Effectiveness of the Cobra aortic catheter for dual-temperature management during adult cardiac surgery. AB - OBJECTIVES: In animals the Cardeon Cobra catheter (Cardeon Corp, Cupertino, Calif) allows independent control of aortic arch and descending aortic temperatures and profoundly reduces cerebral embolization during bypass. This investigation describes the first clinical use of the device during adult cardiac surgery. The purpose of the study was to confirm that the Cobra catheter delivers adequate cerebral and systemic perfusion while providing simultaneous cerebral hypothermia and systemic normothermia during cardiopulmonary bypass. METHODS: In a prospective multicenter study the Cobra aortic catheter was placed in 20 adults undergoing cardiopulmonary bypass. Arch and corporeal temperatures, bypass flows, and arterial blood pressures were recorded intraoperatively. Jugular bulb and mixed venous oxygen saturation was used to assess the adequacy of cerebral and systemic perfusion. RESULTS: Surgeons at 3 institutions placed the Cobra catheter in patients undergoing coronary artery bypass grafting (n = 13), valve (n = 3), and combined valve-bypass (n = 4) operations. Mean total bypass flows of 2.1 +/- 0.2 L x min(-1) x m(-2) maintained mean arterial pressures in arch and descending aortic circulations of greater than 55 mm Hg. A mean differential of 4.3 degrees C between arch and descending aortic temperatures was established before crossclamp application, and a mean maximum temperature differential of 7 degrees C was established during bypass. A 2.4 degrees C temperature differential was maintained at crossclamp removal. Cerebral and systemic venous oxygen saturation remained greater than 65% during bypass. CONCLUSIONS: The Cobra device met all expectations for an arterial cannula with adequate perfusion to the arch and corporeal circulations. Dual perfusion with the Cobra catheter allows for independent temperature control during cardiopulmonary bypass with simultaneous cerebral hypothermia and systemic normothermia. PMID- 12579109 TI - Briefly occlusive coronary anastomosis with tissue adhesive. AB - OBJECTIVE: We assessed the feasibility of a facilitated, briefly occlusive, sutureless coronary anastomosis technique in which side-to-side preglued (octylcyanoacrylate adhesive) bounded walls were opened by a conventional arteriotomy. METHODS: In low-flow (prothrombotic milieu, .05). CONCLUSIONS: The compared methodologies showed similar locations of the ACL insertions, assuring accurate preoperative planning with the CAS system. However, the CAS system requires adjustment to each individual knee anatomy. PMID- 12579149 TI - A review of the function and biomechanics of the meniscofemoral ligaments. AB - PURPOSE: The goal of this study was to review current knowledge on the anatomy, biomechanics, and functions of the meniscofemoral ligaments. TYPE OF STUDY: Literature review. METHODS: A systematic search of the literature spanning the past 2 centuries was performed. This revealed several anatomic and biomechanical studies, which were analyzed for the presence, incidence, function, and biomechanics of the meniscofemoral ligaments. RESULTS: An analysis of 16 anatomic studies revealed that from 1,022 cadaveric knees examined, 931 (91.1%) had at least 1 meniscofemoral ligament; an anterior meniscofemoral ligament was present in 390 (48.2%) of specimens, and a posterior meniscofemoral ligament was present in 569 (70.4%). The 2 ligaments coexisted in 257 knees (31.8%). This high incidence might imply a functional role for these structures. Early theories on the function of these ligaments focused on their role as secondary restraints supplementing the posterior cruciate ligament. More recent hypotheses have concentrated on a role in guiding the motion of the lateral meniscus. CONCLUSIONS: The contribution of the meniscofemoral ligaments in the prognosis and management of posterior cruciate ligament and meniscal injuries remains undetermined. An examination of the structure, properties, and function of the meniscofemoral ligaments reveals that more biomechanical and imaging research is required, together with clinical observations, on the consequences of rupture of these ligaments. PMID- 12579150 TI - Surgical site infections after arthroscopy: Outbreak investigation and case control study. AB - PURPOSE: The goal of this study was to determine the causes of increased post arthroscopy surgical site infections (SSIs) and to define risk factors for infection. TYPE OF STUDY: Outbreak investigation and case control study at a university-affiliated community hospital from 1994 to 1996, with surveillance through 1999. METHODS: Demographic, clinical, and microbiological data were collected on 27 post-arthroscopy SSIs from 1994 through 1999. Risk factors for SSI were identified by case-control analysis and presented as odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Initial investigation revealed an increased annual rate of post-arthroscopy SSIs in 1995 (1.3%). Infection control deficiencies were identified, and feedback was provided to surgeons and staff. Instrument sterilization was standardized, flash sterilization prohibited, and preoperative shaving discouraged. Case-control analysis of 10 cases (from 1994 to 1996) found a statistically significant increase in risk of SSI with intra articular corticosteroid joint injection (OR, 9.33; 95% CI, 1.6 to 64.9); other risk factors did not reach statistical significance. SSI rates dropped after feedback and education (0.34% in 1996). Continued surveillance revealed 2 smaller outbreaks, in December 1997 (1997 rate, 1.13%) and September 1998 (1998 rate, 1.09%). Case-control analysis of the 17 cases occurring in 1997 through 1999 was also performed. The 1997 outbreak appeared to be related to preoperative razor shaving (P =.003), which was then prohibited by hospital policy. One scrub nurse was also associated with 75% of these cases, which were culture-positive for coagulase-negative Staphylococcus. The cases in the 1998 outbreak shared prolonged procedure duration and conversion to arthrotomy. Of 27 cases, 24 required repeat hospitalization and repeat surgery, at an average excess cost of $9,154.84 per case. All received prolonged courses of intravenous or oral antibiotics. CONCLUSIONS: Post-arthroscopy SSIs are associated with significant morbidity and cost. Although small numbers make finding statistical significance difficult in case-control studies, infection control and CDC-recommended interventions can lower SSI rates. Careful definitions, ongoing surveillance, and long-term follow-up are helpful in reporting results of infection control interventions. PMID- 12579151 TI - Healing of osteochondral grafts in an ovine model under the influence of bFGF. AB - PURPOSE: The well-recognized limitations in cartilage healing have lead to the development of a number of resurfacing techniques for defects of joint surfaces. Autologous grafting has developed into 1 of the preferred methods of treatment for focal osteochondral lesions, although basic research on this topic remains sparse. TYPE OF STUDY: In an animal study, questions regarding the healing of osteochondral transplants under the influence of basic fibroblast growth factor (bFGF) were addressed. METHODS: Two osteochondral autologous transplantations (OAT), 1 in the weight-bearing surface of each femoral condyle, were performed on the ovine knee using a standard operative protocol. One of the grafts was bathed in phosphate buffered sulfate (PBS) containing 50 microgram of recombinant human bFGF (rh-bFGF) directly before implantation. Two groups consisting of 10 sheep each were evaluated after 3 and 6 months, respectively. RESULTS: During the evaluation of the specimens, neither osteophytes nor synovial changes were observed. Macroscopically, the superficial border of the transplanted osteochondral plug could easily be outlined at both time periods, even though the cartilage interface of the rh-bFGF specimens was less demarcated. Radiographically, a solid osteointegration of the graft could already be documented at 3 months in the control group. In contrast, integration of the chondral surface of the OAT was not seen macroscopically or microscopically at any point. A firm physical interdigitation of the reconstructed joint surface could not be demonstrated in either of the 2 groups. The augmentation with rh bFGF stimulated the osseous ingrowth, but was not able to improve chondral healing. CONCLUSIONS: The lack of integration of the cartilaginous portion of the transplanted plugs into the reconstructed joint surface, even following augmentation with bFGF, leads to questions regarding the long-term survival of the joint itself. PMID- 12579152 TI - Mechanical testing of absorbable suture anchors. AB - PURPOSE: Absorbable suture anchors offer great advantages but are made of mechanically weak material. The weakest link in the fixation of soft tissue to bone may therefore be the anchor itself. In this study, several commercially available anchors were mechanically tested in vitro. TYPE OF STUDY: Biomechanical bench study. METHODS: Twelve absorbable suture anchor models were implanted into an artificial test bone according to the recommended technique. Testing temperature was 37 degrees C +/- 1 degrees C. The anchors were loaded with an Instron testing machine with the suture material (USP No. 2, Ethibond, Ethicon, Somerville, NJ) in line with the anchor axis, with and without previous abrasion of the suture at the eyelet. Tensile load at failure and failure mode were recorded. To test creep behavior, a permanent load of 100 N was applied to the anchors, and time to failure was recorded. Suture anchor weight and crystallinity were analyzed. RESULTS: Mean failure load on tensile testing using a cross-head speed of 60 mm/min ranged from 124 to 244 N. Failure modes were eyelet failure in 5 cases, suture failure in 6 cases, and anchor pullout in 1 case. In creep testing, eyelet failure occurred in 8 anchor models after a mean duration of 0.5 to 99 hours; 3 anchor models remained intact after 300 hours, and 1 anchor model failed by pullout of the test sample. Crystallinity ranged from 0% (amorphous) to 57.2%; anchor weight ranged from 0.036 to 0.161 g. Mechanical properties did not correlate with crystallinity but with anchor weight. Abrasion of the suture material at the eyelet had little effect on failure load. CONCLUSIONS: At 37 degrees C, structural failure (breaking) of absorbable suture anchors may occur if loaded to the mechanical limit. Absorbable anchors are particularly sensitive to static, long-term loading. PMID- 12579153 TI - A review of ski injuries resulting in combined injury to the anterior cruciate ligament and medial collateral ligaments. AB - Alpine skiing is a global winter recreational sport with 15 million participants in the United States alone, and an overall injury rate of 2.5 per 1,000 ski person-days. Isolated injury to the anterior cruciate ligament (ACL) or the medial collateral ligament (MCL) is common among ski injuries; however, combined injury to these structures is rare. Controversy in the management of ACL instability following alpine ski injury is diminishing with improvements in the techniques of intra-articular cruciate reconstruction. However, the management of the combined ACL-MCL injury remains something of an enigma. Evidence exists to support both surgical and nonsurgical management strategies for the medial structures, but little consensus exists for the timing of the repair. This paper highlights the mechanisms of ski injuries that can result in combined injury to the ACL and MCL. The anatomy and biomechanics of the medial complex as it relates both to stability and operative repair are reviewed, and literature on the techniques and indications used for MCL repair in the setting of a combined injury is presented. On the basis of this review, we believe that an injury to the MCL does not need to be repaired if the ACL is reconstructed after a combined injury. PMID- 12579154 TI - Anterior cruciate ligament reconstruction: A cynical view from the British Isles on the indications for surgery. PMID- 12579155 TI - Arthroscopic treatment of osteochondritis dissecans of the capitellum: Report of 5 female athletes. AB - The management of osteochondritis dissecans of the capitellum of the adolescent elbow is still controversial. We report on 5 cases of female high-level athletes aged from 10 to 19 years (4 gymnasts, 1 waterpolo player). All these athletes had a symptomatic osteochondritis dissecans of the capitellum, which was treated arthroscopically in all cases. Follow-up time averaged 5 months (1 to 6.5 months). During the arthroscopy, loose osteochondral fragments of the capitellum and radial head were removed, and the defect was debrided. Thorough evaluation of the anterior and posterior joint including the olecranon fossa was performed. One of the 5 patients had a loose body requiring arthroscopic removal. Within 6 months after surgery, all except 1 elbow, the elbow with a loose body, regained maximum range of motion. Two patients returned to a high level of gymnastics and 1 was considering return. The short-term results of this treatment suggest that arthroscopic debridement of the loose osteochondral fragments provides a good result. PMID- 12579156 TI - Bipartite patella fracture. AB - Bipartite patella fracture is an uncommon injury that has rarely been described in the literature. It can be quite debilitating in the competitive athlete and is often overlooked by the treating physician. A bone scan can be helpful in confirming the diagnosis, and appropriate treatment often results in a successful outcome. PMID- 12579157 TI - Arthroscopic resection of volar ganglion of the wrist: A new technique. AB - We evaluated a new arthroscopic technique for treating volar ganglion of the wrist. Arthroscopic resection of volar wrist ganglion was performed under local anesthesia to the portal sites. Arthroscopic instruments were introduced dorsally, and the intra-articular lesion at the interval between the radioscaphocapitate (RSC) and long radiolunate (LRL) ligaments was debrided under direct view. Mucinous content of the ganglion was evacuated using suction, but the ganglion sac was not removed. The technique was successful in 5 cases between August 1997 and November 1998, with an average surgery time of 55 minutes. No complications or recurrence occurred at a mean follow-up of 16.4 months. This is the first report of arthroscopic decompression of volar wrist ganglions and provides new insight into the etiology of these lesions. PMID- 12579158 TI - Osteochondritis dissecans of the capitellum: Arthroscopic-assisted treatment of large, full-thickness defects in young patients. AB - Methods of treatment for mild juvenile osteochondritis dissecans (OCD) of the capitellum include activity modification and periods of immobilization, followed by therapy and range of motion exercises. Surgical treatment may be indicated if there are persistent symptoms, a loose body, the child is approaching skeletal maturity, or if magnetic resonance imaging reveals a fracture through the articular cartilage. We describe a technique of arthrotomy, using an arthroscope as a visual aid, for the treatment of large, full-thickness OCD lesions of the capitellum in young patients. The arthroscopic camera allows the surgeon to view difficult-to-see areas, and photographs can be taken throughout the procedure for documentation. PMID- 12579160 TI - The prevalence of ocular disease in chronic lymphocytic leukaemia. PMID- 12579159 TI - Meeting the challenge of glaucoma after paediatric cataract surgery. PMID- 12579161 TI - Is review of enriched populations the way forward for glaucoma case detection? PMID- 12579162 TI - 'Non-medical' prescribing in glaucoma. PMID- 12579163 TI - The British Ophthalmological Surveillance Unit: an evaluation of the first 3 years. AB - We report an evaluation of the British Ophthalmological Surveillance Unit (BOSU), a nationwide active surveillance scheme to assist with the case ascertainment of rare eye conditions. The evaluation assessed participation rates, study applications and research outputs. In addition, through an anonymous postal survey we ascertained the opinions of ophthalmologists regarding the usefulness, levels of feedback, barriers to participation and levels of case ascertainment. Over the first 3 years, the 4-month mean participation rate has improved from 58% to 71%. Ten studies have used this system for case ascertainment. To date three journal publications and 10 conference presentations have reported findings from these studies. It was observed that 582/870 (68%) questionnaires were returned by ophthalmologists, of whom 95% considered BOSU as very or quite useful. In all, 71% reported overall feedback to be sufficient. However, 34% requested greater feedback from research groups. Reported barriers to effective participation were 'having to remember the patient's identity' (52%) and paying for return postage of cards (22%). However, 72% of respondents did not consider the work involved in reporting a case to be prohibitive. Self-reported levels of case ascertainment by ophthalmologists for completed studies ranged between 72% and 95%. This population-based surveillance system provides an effective method for prospective case identification and subsequent data collection. It enables the study of sufficiently representative samples to allow meaningful epidemiological analysis and avoid bias. Its success relies upon the high level of support that it currently receives from ophthalmologists. PMID- 12579164 TI - Phacoemulsification and intraocular lens implantation following pars plana vitrectomy: a prospective study. AB - AIMS: To examine the intraoperative characteristics of phacoemulsification in eyes that had undergone pars plana vitrectomy and to determine complication rates and visual outcomes. METHODS: A prospective study of 45 consecutive cataract operations on vitrectomised eyes performed over a 9-month period by a single surgeon. Data were collected on preoperative characteristics, intraoperative observations and complications, postoperative complications, and visual acuities. RESULTS: Zonular laxity with abnormal deepening of the anterior chamber and mydriasis was noted in six eyes that had undergone extensive vitreous removal for the treatment of retinal detachment or proliferative diabetic retinopathy, but not in eyes that had had limited 'core vitrectomy' such as for macular hole or epiretinal membrane. In two eyes, the initial deepening of the anterior chamber was accompanied by paradoxical shallowing with miosis (the 'infusion deviation syndrome'). Sight-threatening complications were rare. Snellen visual acuity improved in 84.6% of eyes previously treated for 'macular hole, 85.7% of eyes treated for 'macula-on' retinal detachment, 66.7% of eyes treated for 'macula off' retinal detachment, and 57.1% of eyes treated for diabetic retinopathy. CONCLUSIONS: Surgeons need to be aware of additional challenges arising from zonular instability in eyes that have had extensive vitreous removal. A lower infusion bottle height may guard against sudden changes in anterior chamber depth. The use of a corneal tunnel avoids difficulties associated with an unusually deep anterior chamber. Visual acuity is less likely to improve in patients previously treated for diabetic retinopathy or for macula-off retinal detachment. PMID- 12579165 TI - Community refinement of glaucoma referrals. AB - AIM: To describe a Manchester-based glaucoma referral refinement scheme designed to reduce the number of false-positive referrals to the hospital eye service. To report on the first years results of this scheme and its financial costs to the NHS. METHODS: Patients with suspected glaucoma, instead of being referred to their GP and then on to the hospital eye service, were referred to a group of specially trained community optometrists working to an agreed set of referral criteria. Those patients who did not meet the referral criteria were returned to the referring optometrist, while those who met the referral criteria were referred directly to Manchester Royal Eye Hospital. The patient's GP was informed in all cases. RESULTS: The number of suspect glaucoma cases referred to the Manchester Royal Eye Hospital was reduced by 40%. This figure is close to the percentage of false-positive referrals measured at Manchester Royal Eye Hospital prior to the onset of this study. The information accompanying referral has been improved and the scheme produces a small financial cost saving to the NHS of approximately 17 pounds sterling per patient. CONCLUSION: Community refinement of suspect glaucoma offers some important benefits over the current referral pathway. PMID- 12579166 TI - The prevalence of ocular disease in chronic lymphocytic leukaemia. AB - AIM: Previous surveys of ocular disease in leukaemia patients have shown the retina, choroid, and orbit as the most commonly involved sites. Depending on the type of leukaemia and the study design, the prevalence of ocular disease in leukaemia varies from 30 to 90%. Although chronic lymphocytic leukaemia (CLL) is the most common leukaemia in Western countries, the prevalence of ocular disease in CLL is not known. The aim of this prospective study was to estimate the prevalence of ocular disease in CLL. PATIENTS AND METHODS: All CLL patients attending either a teaching or district general hospital haematology clinic were invited to undergo a full eye examination. The clinical stage of the CLL (Binet) at the time of the eye examination and the most recent full blood count (FBC) indices were recorded for each patient. RESULTS: Over 6 months, 25 patients with an average age of 65 were recruited. A total of 18 patients had Binet stage A disease, three had stage B, and four stage C. The mean FBC indices at the time of examination were haemoglobin 13.5 g/dl, white cell count 37.4 x 10(9)/l, and platelets 172 x 10(9)/l. Only three patients (12%) had ocular disease that was likely to be a secondary complication of CLL. These secondary complications included bilateral posterior subcapsular cataract following radiotherapy, unilateral acute retinal necrosis, and unilateral conjunctival vascular anomalies. The presence of ocular complications was not related to the CLL stage or to the current FBC parameters. DISCUSSION: This study demonstrates that ocular involvement in CLL is uncommon, a reflection of the indolent course of CLL as compared to other leukaemias. Although sight-threatening ocular complications have been documented in this and other studies, the low prevalence of complications and the lack of association with disease parameters make it impossible to identify a 'high-risk' group. Routine screening of CLL patients for ocular complications is not justified. PMID- 12579167 TI - Association of tumour necrosis factor alpha -308 gene polymorphism with primary open-angle glaucoma in Chinese. AB - PURPOSE: Genetic factors are known to play a role in the aetiology of glaucoma, and in particular the role of the immune system is highly suspected. In this study, we evaluated the association between tumour necrosis factor alpha -308 (TNF alpha -308) and primary open-angle glaucoma (POAG). METHODS: A total of sixty POAG patients and 103 healthy volunteers as control group were enrolled in this case-controlled study. Furthermore, we used polymerase chain reaction based analysis to resolve the TNF alpha -308 polymorphism. Statistical analysis for the relative risk of TNF alpha -308 polymorphism was compared by the chi(2) test. RESULTS: There were significant differences in the distribution of the polymorphism between the POAG patients and the control subjects (P = 0.00016; P < 0.05) and it was found that the A(-308) allele occurred more frequently in POAG patients (odds ratio: 2.72; 95% confidence interval: 1.66-4.45). CONCLUSION: The results of our study concluded that the distribution of TNF alpha -308 was significantly higher in the POAG patients than in the control group. Therefore, the A(-308) allele appears to be associated with POAG and, therefore, could be used as a genetic marker for disease mapping. POAG is a complex disease, and a single gene could not be responsible. Understanding the role of genetic polymorphisms, like TNF alpha, could be a prediction of the disease and useful for developing new treatments for POAG. PMID- 12579168 TI - Psychosocial distress associated with disfiguring eye conditions. AB - PURPOSE OF STUDY: Visible disfigurement is associated with a range of psychosocial difficulties; however, the problems encountered by patients with eye conditions are under-researched. This study aimed to establish the extent and type of psychosocial distress in this patient population, and to explore the extent to which patients' needs are currently met. METHOD: Standardised measures (anxiety, depression, appearance-related distress, and quality of life) and a semistructured interview (exploring individual experiences and satisfaction with care) were administered to 153 consecutive attenders at outpatient eye clinics in three different hospitals. Responses were analysed using descriptive statistics, multiple regression, and cluster analyses. RESULTS: Between 10 and 49% of the patient sample scored unfavourably on standardised measures in comparison with normative values. Cluster analyses revealed that more distressed patients typically exhibited higher levels of anxiety, depression, social anxiety, self consciousness, and social avoidance. Quality of life scores were also less favourable. Distressed patients felt less well informed, less involved in their treatment, and less well supported in the clinic setting. Variability within the sample was high; however, the effects of the cause and duration of the condition were not significantly related to distress. Clinic staff identified a variety of constraints to meeting patients' need, including patient numbers, the lack of appropriate training, and the lack of referral possibilities. CONCLUSIONS: A considerable proportion of patients with disfiguring eye conditions experience high levels of psychosocial distress. Several options exist for ways in which patients' needs might more effectively be met within the constraints currently facing care providers. PMID- 12579169 TI - The efficacy and safety of the timolol/dorzolamide fixed combination vs latanoprost in exfoliation glaucoma. AB - PURPOSE: To evaluate the safety and efficacy of the timolol/dorzolamide fixed combination vs latanoprost 0.005% in exfoliation glaucoma patients. METHODS: We randomized in an observer-masked fashion 65 newly diagnosed exfoliation glaucoma patients to either the timolol/dorzolamide twice daily or latanoprost daily treatment for 2 months and then crossed these over to the other treatment. RESULTS: A total of fifty-four patients completed the study. After 2 months of chronic dosing, the morning intraocular pressure (IOP) (10:00) was reduced from a baseline of 31.2+/-6.5 mmHg to 18.1+/-3.0 with the fixed combination and to 18.9+/-4.1 mmHg with latanoprost (P = 0.21). Six patients were discontinued early from both treatment periods owing to inadequate IOP control and two others were discontinued from latanoprost treatment only. The fixed combination showed a significantly greater incidence of taste perversion (P < 0.001) and stinging upon instillation (P = 0.036), while latanoprost showed a trend for increased conjunctival injection (P = 0.056). However, five patients demonstrated either bradycardia or asthmatic symptoms with initiation of the fixed combination therapy. One patient on latanoprost complained of dizziness. Patient preference was generally given to latanoprost (63 vs 20.3%) mainly because of its once daily dosing (P < 0001). CONCLUSIONS: This study suggests that both latanoprost and the timolol/dorzolamide fixed combination are efficacious in the treatment of newly diagnosed exfoliation glaucoma. PMID- 12579170 TI - Nurse prescribing in glaucoma. AB - AIMS/PURPOSE: To investigate a nurse-led assessment clinic of new referrals of possible glaucoma. To determine the potential for nurse prescribing. To review the background literature about nurse prescribing. METHODS: Audit of the outcomes of patients attending the nurse-led glaucoma assessment clinic during two defined periods (169 patients). An audit of all patients started on timolol or latanoprost treatment by a nurse following a protocol. RESULTS: A total of 46 patients were commenced on treatment at the clinic, 31 on timolol, 14 on latanoprost, and one on brimonidine. Four of these had the treatment stopped at the review clinic in order to reassess the diagnosis. Four patients in the timolol group developed side effects requiring a change in medication, but these could not have been predicted from their past medical history. Nine patients had treatment changed or added to because the intraocular pressure was felt to be inadequately controlled. During the two 3-month audits, a further 11 patients were commenced on treatment for glaucoma at the review clinic. CONCLUSIONS: Initial data from this clinic suggest that nurses possess the diagnostic skills necessary to prescribe for new glaucoma patients. The legal and administrative frame works are developing for more nurses to be able to prescribe. With the newer prostaglandin treatments for glaucoma being available, nurses may usefully and safely be able to prescribe first-line treatments for glaucoma. PMID- 12579171 TI - Trabeculectomy with or without mitomycin-C for paediatric glaucoma in aphakia and pseudophakia following congenital cataract surgery. AB - PURPOSE: To evaluate the safety and efficacy of trabeculectomy with or without mitomycin-C (MMC) in the management of glaucoma in aphakia and pseudophakia following congenital cataract surgery. PATIENTS AND METHODS: All patients of glaucoma with aphakia or pseudophakia who underwent trabeculectomy with or without MMC from January 1989 to April 2000 were included. The medical records of 19 consecutive patients (23 eyes) were reviewed. Data collected from a retrospective chart review were analysed. Outcome measures were evaluated using Kaplan-Meier survival analysis. Pre- and postoperative intraocular pressures (IOPs), visual acuities, success rate, bleb characteristics, surgical failure and complications were the main outcome measures. Successful IOP control was defined as an IOP between 6 and 21 mmHg, without antiglaucoma medications, without further antiglaucoma surgery and without any sight-threatening complication. RESULTS: The mean age of patients was 8.8+/-5.5 years at the time of trabeculectomy with MMC compared to 11.0+/-12.4 years for trabeculectomy without MMC. Eight patients underwent trabeculectomy with MMC and 11 patients underwent trabeculectomy without MMC. There was no statistically significant difference between the two groups in terms of visual acuity, IOP, antiglaucoma medications, age at cataract surgery and at trabeculectomy. The IOP reduced from a preoperative level of 34.2+/-8.9 mmHg (range: 20-52) to a postoperative level of 18.4+/-12.2 mmHg (range: 2-60) with a mean follow-up of 24.2+/-17.9 months. The mean reduction in IOP in the MMC group was 15.5+/-17.3 and 16.3+/-13.8 mmHg in the other group (P = 0.967). Overall, complete success was achieved in 36.8%, qualified success in 21.1% and surgical failure in 42.1% of patients with a mean follow-up of 24.2+/-17.9 months. There was no difference in the success between the two groups at the last follow-up. One patient developed bleb-related endophthalmitis in both eyes following trabeculectomy with MMC. CONCLUSIONS: The success rate of trabeculectomy in glaucoma following congenital cataract surgery was 36.8% at the end of 3 years. The present study proves a poor success rate of trabeculectomy in a small series of aphakic Asian Indian patients even with the use of MMC. PMID- 12579172 TI - Ethnic differences in the demand incidence of retinal detachments in two districts in the West Midlands. AB - AIM: To evaluate the epidemiological characteristics of rhegmatogenous retinal detachments (RRDs) in two defined populations in the West Midlands of England. METHODS: A 5-year retrospective study of patients who underwent surgical repair of their RRD between April 1994 and March 1999. Statistical analysis was done by the Statistical Program for Social Sciences (SPSS) version 10.0. RESULTS: The total demand incidence of RRD was 11.3 and 6.3 per 100,000 in Wolverhampton and Walsall, respectively. The Asian subgroup had a low demand incidence of 4.6 and 2.0 per 100,000 in each area, respectively. In Wolverhampton the highest age specific demand incidence was in the 70 to 79-year age group at 29.1 per 100,000, whereas in Walsall it was 98.6 per 100,000 in the 85+ age group. Nontraumatic phakic detachments had the highest demand incidence of 9.7 per 100,000, whereas nontraumatic pseudophakic and aphakic R/D were much lower at 1.2 and 0.3 per 100,000, respectively. There was a 1 : 1.5 female-to-male ratio, with a mean age of 57.8 years (95% CI 54.7-61.0) in females and 55.5 years (95% CI 53.1-57.9) for males. A total of 41.6% (142) of patients presented initially to the Wolverhampton Eye Infirmary Accident and Emergency Department (A&E). CONCLUSION: This is the first UK-based study. The demand incidence in Caucasians is similar to worldwide figures. Asians have a three times lower incidence of retinal detachments. The demand incidence increases with age. Females have a higher mean age than males. Males (89.5%) were more likely to suffer from traumatic detachments. Younger patients were more likely to present to the optometrist initially. Less than half of the patients will present initially to the eye A&E. PMID- 12579173 TI - Virtual reality simulator for vitreoretinal surgery. AB - AIM: To develop computer simulation of steps in vitreoretinal surgery using virtual reality technology. MATERIAL AND METHODS: A workstation with three dimensional position tracking stylus was attached to a Pentium II desktop PC with a graphic accelerator. Computer algorithms were developed using Open GL and Microsoft Visual C++ languages to control the interaction and update the visual feedback tracking the instruments. Soft tissue computer modelling was carried out to mimic the removal of a vitreous opacity. Lens touch with the instruments was also detected. Mathematical modelling to allow for lens distortion was taken into account. RESULTS: A virtual reality computer model has been developed that can simulate initial steps of vitreoretinal surgery. Soft tissue modelling of the vitreous opacity and its removal by the vitrector was successfully simulated. The movements of the active and passive instruments in the dummy eye corresponded to the movements on the computer screen. On evaluation of the system, there was a minimal but discernable time lag between the stylus movement and the visual feedback. There was no tactile feedback when the lens touch was simulated. No further complex vitreoretinal surgery simulation was possible at this stage. PMID- 12579174 TI - Effectiveness of oxygenation and suction in cataract surgery: is suction of CO2 enriched air under the drape during cataract surgery necessary? AB - PURPOSE: To investigate the effectiveness of aspiration of expired air by a suction system on peripheral oxygen saturation (SpO(2)) and end tidal carbon dioxide (EtCO(2)) during cataract surgery. MATERIALS AND METHODS: In total, 160 pre-medicated patients aged between 40 and 70 years (ASA I-III, classification of patients according to the American Society of Anesthesiologists) scheduled for cataract surgery under retrobulbar or peribulbar block were examined in a randomised, single-blind manner. The patients were sedated with 3 mg midazolam i.v. 15 min before operation and were monitored with an anaesthesia monitor. Heart rate (HR), non-invasive mean arterial pressure (MAP), SpO(2) and EtCO(2) were continuously measured using a standardised monitor. The first group (non suction group, n = 80) received 4 L min(-1) O(2) with nasal cannule while the second group (suction group, n = 80) received 4 L min(-1) O(2) with nasal cannule, and the expired air was aspired with a Y-piece suction system. EtCO(2) was measured with the line of the sampling tube in the anaesthesia monitor. Respiratory rate (RR) was counted for a period of 1 min at each measurement time with thoracic excursions. The results were evaluated by unpaired t-test and analysis of variance. RESULTS: Severe reduction of SpO(2) and raising of EtCO(2) were observed significantly in the first group during the operation. RR, HR and MAP increased due to hypoxaemia. In the second group, SpO(2) was stabilised at high levels and EtCO(2) did not increase. RR, HR and MAP levels remained within the normal limits. Differences between the two groups were statistically significant (P < 0.05). CONCLUSION: During cataract surgery with local anaesthesia, SpO(2) decreases and RR, HR and MAP increase because of reinspiration of expired air under the drape. Insufflation of O(2) and aspiration of expired air with a suction system have prevented severe reduction of SpO(2) and raising of EtCO(2). It was suggested that O(2) delivery and use of an aspiration system decreased the risk of hypoxaemia significantly in the patients undergoing the cataract surgery. PMID- 12579175 TI - Correlation of lipid layer thickness measurements with fluorescein tear film break-up time and Schirmer's test. AB - PURPOSE: This study correlates measurement of lipid layer thickness (LLT) with two frequently used dry eye tests, fluorescein break-up time (FBUT) and Schirmer's test with anaesthesia (STA). METHODS: Subjects (n = 44 eyes) with symptoms of dry eye and positive results for dry eye with either FBUT or STA or both were selected. Quantification of LLT was performed by the observation of colour interference patterns in zones of specular reflection using a custom designed instrument. RESULTS: All correlations among pairs of tests were strong and exhibited a significance of P < 0.000: STA with FBUT, Pearson's correlation 0.653; STA with LLT, 0.764; FBUT with LLT, 0.751. When LLT was high, ie > or = 120 nm, which occurred in 14 eyes, STA was also elevated in those eyes and FBUT was high in 13 of the 14 eyes. When LLT was low, ie < or = 60, which occurred in 22 eyes, STA was below normal in 14 of the 22 eyes, and FBUT was below normal in 15 of the 22 eyes. These clinical observations paralleled the statistical findings computed from the entire data set. CONCLUSIONS: The correlations demonstrated in this study support the premise (1) that measurement of LLT is a reliable test for the diagnosis of dry eye, and (2) that aqueous deficiency and lipid deficiency, as they apply to dry eye disorders, are not mutually exclusive. PMID- 12579176 TI - Choroidal blood flow and arterial blood pressure. AB - PURPOSE: Untreated hypertension is associated with ocular complications and is a risk factor for the development and progression of vascular ocular pathologies. We set out to investigate the association between systemic blood pressure and choroidal blood flow. METHODS: All subjects were male non-smokers, who did not receive any medication and had normal or slightly elevated blood pressure (systolic blood pressure < or = 160 mmHg; diastolic blood pressure < or = 100 mmHg). The association between systemic blood pressure and fundus pulsation amplitude, a measure of pulsatile choroidal blood flow, was investigated in 318 volunteers. In addition, the association between systemic blood pressure and blood flow velocities in the posterior ciliary arteries supplying the choroid was investigated in these subjects. RESULTS: Ocular fundus pulsation amplitude (r = 0.252; P < 0.001) and mean flow velocity in the posterior ciliary arteries (r = 0.346, P < 0.001) were significantly associated with mean arterial pressure. The correlation of ocular haemodynamic variables with systolic and diastolic blood pressure was in the same range. CONCLUSIONS: Our data indicate a small, but significant increase in choroidal blood flow with increasing blood pressure. PMID- 12579177 TI - Sight and comfort: complex procedures in end-stage Stevens-Johnson syndrome. AB - BACKGROUND: We describe our complex surgical techniques in the management of a patient with end-stage ocular surface disease from Stevens-Johnson syndrome. METHODS AND RESULTS: Her severe discomfort due to absolute ocular dryness in the right eye was successfully treated with submandibular gland autotransplantation. Impending loss of the left eye due to repeated perforation and infection was prevented with a penetrating corneal graft covered by a new modification of a Cutler-Beard-type full-thickness lower lid skin advancement-flap. Visual rehabilitation was achieved by means of osteo-odonto-keratoprosthesis. CONCLUSION: The procedures described allow the preservation of eyes doomed for enucleation, relief from severe discomfort and rehabilitation from corneal blindness. Due to their complex nature they require the collaborative subspecialist surgical skills of opthalmologists and maxillofacial surgeons. PMID- 12579178 TI - Giant cell arteritis with a normal ESR and CRP. PMID- 12579179 TI - The role of an ophthalmologist in suspected non-accidental injury. PMID- 12579180 TI - Familial myelinated retinal nerve fibres. PMID- 12579181 TI - Post-chemotherapy premacular subhyaloid haemorrhage. PMID- 12579182 TI - Uveitis in a patient with common variable immunodeficiency. PMID- 12579183 TI - Mucucoele of the maxillary sinus and the eye. PMID- 12579184 TI - Aeromonas sobria corneal ulcer. PMID- 12579185 TI - Orbital cellulitis after peribulbar anaesthesia for cataract surgery. PMID- 12579186 TI - Self-inflicted anterior scleritis. PMID- 12579187 TI - Sanichlor-induced atopic dermatitis and asthma in ophthalmologists. PMID- 12579188 TI - Central retinal artery occlusion following staphylococcal orbital cellulitis. PMID- 12579190 TI - Exercise-related visual loss in patients with glaucoma. PMID- 12579192 TI - Nd:YAG vitreolysis as a treatment for vitreous floaters. PMID- 12579195 TI - The thrombomodulin-protein C system is essential for the maintenance of pregnancy. AB - Disruption of the mouse gene encoding the blood coagulation inhibitor thrombomodulin (Thbd) leads to embryonic lethality caused by an unknown defect in the placenta. We show that the abortion of thrombomodulin-deficient embryos is caused by tissue factor-initiated activation of the blood coagulation cascade at the feto-maternal interface. Activated coagulation factors induce cell death and growth inhibition of placental trophoblast cells by two distinct mechanisms. The death of giant trophoblast cells is caused by conversion of the thrombin substrate fibrinogen to fibrin and subsequent formation of fibrin degradation products. In contrast, the growth arrest of trophoblast cells is not mediated by fibrin, but is a likely result of engagement of protease-activated receptors (PAR)-2 and PAR-4 by coagulation factors. These findings show a new function for the thrombomodulin-protein C system in controlling the growth and survival of trophoblast cells in the placenta. This function is essential for the maintenance of pregnancy. PMID- 12579196 TI - Eradication of systemic B-cell tumors by genetically targeted human T lymphocytes co-stimulated by CD80 and interleukin-15. AB - The genetic transfer of antigen receptors provides a means to rapidly generate autologous tumor-reactive T lymphocytes. However, recognition of tumor antigens by cytotoxic T cells is only one step towards effective cancer immunotherapy. Other crucial biological prerequisites must be fulfilled to expand tumor-reactive T cells that retain a functional phenotype, including in vivo cytolytic activity and the ability to travel to tumor sites without prematurely succumbing to apoptosis. We show that these requirements are met by expanding peripheral blood T cells genetically targeted to the CD19 antigen in the presence of CD80 and interleukin-15 (IL-15). T cells expanded in the presence of IL-15 uniquely persist in tumor-bearing severe combined immunodeficiency (SCID)-Beige mice and eradicate disseminated intramedullary tumors. Their anti-tumor activity is further enhanced by in vivo co-stimulation. In addition, transduced T cells from patients with chronic lymphocytic leukemia (CLL) effectively lyse autologous tumor cells. These findings strongly support the clinical feasibility of this therapeutic strategy. PMID- 12579197 TI - RNA interference targeting Fas protects mice from fulminant hepatitis. AB - RNA interference (RNAi) is a powerful tool to silence gene expression post transcriptionally. However, its potential to treat or prevent disease remains unproven. Fas-mediated apoptosis is implicated in a broad spectrum of liver diseases, where inhibiting hepatocyte death is life-saving. We investigated the in vivo silencing effect of small interfering RNA (siRNA) duplexes targeting the gene Fas (also known as Tnfrsf6), encoding the Fas receptor, to protect mice from liver failure and fibrosis in two models of autoimmune hepatitis. Intravenous injection of Fas siRNA specifically reduced Fas mRNA levels and expression of Fas protein in mouse hepatocytes, and the effects persisted without diminution for 10 days. Hepatocytes isolated from mice treated with Fas siRNA were resistant to apoptosis when exposed to Fas-specific antibody or co-cultured with concanavalin A (ConA)-stimulated hepatic mononuclear cells. Treatment with Fas siRNA 2 days before ConA challenge abrogated hepatocyte necrosis and inflammatory infiltration and markedly reduced serum concentrations of transaminases. Administering Fas siRNA beginning one week after initiating weekly ConA injections protected mice from liver fibrosis. In a more fulminant hepatitis induced by injecting agonistic Fas-specific antibody, 82% of mice treated with siRNA that effectively silenced Fas survived for 10 days of observation, whereas all control mice died within 3 days. Silencing Fas expression with RNAi holds therapeutic promise to prevent liver injury by protecting hepatocytes from cytotoxicity. PMID- 12579198 TI - Prevention of virus transmission to macaque monkeys by a vaginally applied monoclonal antibody to HIV-1 gp120. AB - A topical microbicide reduces the probability of virus transmission when applied to the vagina or rectum of a person at risk of sexually acquiring HIV-1 infection. An effective microbicide could significantly reduce the global spread of HIV-1, particularly if women were able to use it covertly to protect themselves. A microbicide could target the incoming virus and either permanently inactivate it or reduce its infectivity, or it could block receptors on susceptible cells near the sites of transmission. We describe here how vaginal administration of the broadly neutralizing human monoclonal antibody b12 can protect macaques from simian-human immunodeficiency virus (SHIV) infection through the vagina. Only 3 of 12 animals receiving 5 mg b12 vaginally in either saline or a gel and then challenged vaginally (up to 2 h later) with SHIV-162P4 became infected. In contrast, infection occurred in 12 of 13 animals given various control agents under similar conditions. Lower amounts of b12 were less effective, suggesting that protection was dose dependent. These observations support the concept that viral entry inhibitors can help prevent the sexual transmission of HIV-1 to humans. PMID- 12579199 TI - NTP Toxicology and Carcinogenesis Studies of Phenolphthalein (CAS No. 77-09-8) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Phenolphthalein is used as a laboratory reagent and acid-base indicator and in over-the-counter laxative preparations. The National Cancer Institute nominated phenolphthalein for study because of its widespread use as a component in numerous laxative preparations and the lack of adequate testing for carcinogenicity in experimental animals. Male and female F344/N rats and B6C3F1 mice were exposed to phenolphthalein (98% to 99% pure) in feed for 14 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and mouse peripheral blood. 14 DAY STUDY IN RATS: Groups of five male and five female F344/N rats were given 0, 6,250, 12,500, 25,000, 50,000, or 100,000 ppm phenolphthalein in feed for 14 days. All rats survived to the end of the study. The final mean body weights of all exposed groups of rats were similar to those of the controls. No chemical related gross or microscopic lesions were observed. 14-DAY STUDY IN MICE: Groups of five male and five female B6C3F1 mice were given 0, 6,250, 12,500, 25,000, 50,000, or 100,000 ppm phenolphthalein in feed for 14 days. All mice survived to the end of the study. The final mean body weights of all exposed groups of mice were similar to those of the controls. No chemical-related gross or microscopic lesions were observed. 13-WEEK STUDY IN RATS: Groups of 10 male and 9 or 10 female F344/N rats were given 0, 3,000, 6,000, 12,000, 25,000, or 50,000 ppm phenolphthalein (equivalent to average daily doses of approximately 200, 400, 800, 1,600, or 3,500 mg phenolphthalein/kg body weight to males and 200, 400, 800, 1,700, or 3,600 mg/kg to females) in feed for 13 weeks. Additional groups of 10 male and 10 female rats designated for clinical pathology evaluations were also given 0, 3,000, 6,000, 12,000, 25,000, or 50,000 ppm phenolphthalein in feed until day 21. All core study rats survived to the end of the study. The final mean body weight of the 50,000 ppm females and the body weight gains of the 25,000 and 50,000 ppm females were significantly lower than those of the controls. The final mean body weights and mean body weight gains of all other exposed groups were similar to those of the controls. There was no cathartic action or any other clinical finding attributed to exposure to phenolphthalein. The few differences in the hematology and clinical chemistry parameters were sporadic and were not considered to be chemical related. The percentage of motile sperm in the 12,000 ppm males was significantly greater than that in the controls, but no other significant differences in sperm morphology or vaginal cytology between exposed and control groups were observed. Absolute and relative liver weights of 25,000 and 50,000 ppm males were significantly greater than those of the controls. No chemical-related gross or microscopic lesions were observed. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were given 0, 3,000, 6,000, 12,000, 25,000, or 50,000 ppm phenolphthalein (equivalent to average daily doses of approximately 500, 1,000, 2,000, 4,100, or 9,000 mg phenolphthalein/kg body weight to males and 600, 1,200, 2,400, 5,000, or 10,500 mg/kg to females) in feed for 13 weeks. All mice survived until the end of the study. The final mean body weights and mean body weight gains of all exposed groups were similar to those of the controls. There was no cathartic action or any other clinical finding attributed to exposure to phenolphthalein. The absolute right cauda weight of the 12,000 ppm males and the absolute right epididymis weights of 12,000, 25,000, and 50,000 ppm males were significantly less than those of the controls. The percentages of abnormal sperm in 12,000, 25,000, and 50,000 ppm males were significantly greater than that in the control group, and the sperm concentrations in 12,000 and 50,000 ppm males were significantly less than that of the control group. The absolute and relative right testis weights of males exposed to 6,000 ppm or greater and the absolute right testis weight of 3,000 ppm mof 3,000 ppm males were significantly less than those of the controls. The incidences of hypoplasia of the bone marrow in males and females exposed to 12,000 ppm or greater were significantly greater than those in the controls. The incidences of hematopoiesis of the spleen in 25,000 and 50,000 ppm males were significantly greater than that in the controls. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were given 0, 12,000, 25,000, or 50,000 ppm phenolphthalein (equivalent to average daily doses of approximately 500, 1,000, or 2,000 mg phenolphthalein/kg body weight to males and 500, 1,000, or 2,500 mg/kg to females) in feed for 2 years. Survival, Body Weights, and Clinical Findings: Survival of exposed males and females was similar to that of the controls. The mean body weights of exposed males were less than those of the controls through most of the second year of the study, and the mean body weights of exposed females were less than those of the controls from about week 16 until the end of the study. Clinical findings attributed to phenolphthalein exposure included thin appearance and ruffled fur in all exposed groups of males. Determinations of Total Phenolphthalein in Plasma: The mean plasma concentrations of total phenolphthalein (free and conjugated) after 2 years of exposure varied little with time of day. Plasma concentrations of total phenolphthalein were approximately the same between exposure groups and between males and females. Pathology Findings: The incidences of benign pheochromocytoma of the adrenal medulla in all exposed groups of males were significantly greater than those in the controls and occurred with a significant positive trend. The incidences of benign pheochromocytoma in 12,000 ppm females and of benign or malignant pheochromocytoma (combined) in 12,000 and 25,000 ppm females were significantly greater than those in the controls. The numbers of exposed males with bilateral benign pheochromocytomas exceeded the number of controls with these neoplasms. The incidences of malignant pheochromocytomas in exposed rats were similar to those in the controls. The incidences of focal hyperplasia of the adrenal medulla in the 12,000 and 50,000 ppm males were significantly greater than in the controls. The incidences of renal tubule adenoma in 50,000 ppm male rats and of renal tubule adenoma or carcinoma (combined) in 12,000 and 50,000 ppm male rats were significantly greater than those in the controls. Although the increased incidences were predominantly of renal tubule adenoma, four carcinomas were observed in exposed males (0 ppm, 0/50; 12,000 ppm, 1/50; 25,000 ppm, 1/50; 50,000 ppm, 2/50). The incidences of renal tubule neoplasms in exposed groups of females were similar to those in the controls. The findings from an extended evaluation (step section) of the kidneys of female rats were similar to those from the standard evaluation. The incidences of nephropathy in all exposed groups of females were significantly greater than in the controls, and the severity of nephropathy in all exposed groups of males and in 25,000 and 50,000 ppm females was significantly greater than in the controls. The incidences of diffuse hyperplasia of the parathyroid gland (0/41, 16/48, 14/49, 14/46), fibrous osteodystrophy of the bone (0/50, 17/50, 14/50, 12/50), and mineralization (0/50, 11/50, 5/50, 5/49) and degeneration (0/50, 11/50, 5/50, 4/49) of the glandular stomach in exposed groups of males were generally significantly greater than those in the controls. The incidences of hyperplasia of the thyroid gland C-cells (13/50, 3/50, 9/49, 4/49) in 12,000 and 50,000 ppm males were significantly less than in the controls. These lesions are commonly observed in male rats with more advanced nephropathy and are considered to be associated with a calcium/phosphorus imbalance created by compromised functional capacity of the kidney. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were given 0, 3,000, 6,000, or 12,000 ppm phenolphthalein (equivalent to average daily doses of approximately 300, 600, or 1,200 mg phenolphthalein/kg body weight to males and 400, 800, or 1,500 mg/kg to females) in feed for 2 years. Survival, Body Weights, and Clinical Findings: Survival of the 12,000 ppm females was significantly lower than that of the controls; survival of all other exposed groups of mice was similar to that of the controls. The mean body weights of 12,000 ppm males were slightly less than those of the controls beginning at week 93 of the study, and the mean body weights of the 3,000, 6,000, and 12,000 ppm females were less than those of the controls during most of the second year of the study. In exposed mice, there were no clinical findings related to phenolphthalein exposure. Determinations of Total Phenolphthalein in Plasma: The mean plasma concentrations of total phenolphthalein (free and conjugated) after 2 years of exposure varied little with time of day. Plasma concentrations of total phenolphthalein were approximately the same between exposure groups and between males and females. Pathology Findings: The incidences of histiocytic sarcoma in 6,000 and 12,000 ppm males and females were significantly greater than those in the controls and occurred with a significant positive trend. In this study, histiocytic sarcoma was consistently observed in the liver with several other sites (e.g., spleen, lung, bone marrow, and various lymph nodes) involved less frequently. The incidences of all types of malignant lymphoma and of lymphoma of thymic origin in all exposed groups of females were significantly greater than those in the controls and occurred with significant positive trends, while the incidences of all types of malignant lymphoma in all exposed groups of males were similar to that in the controls. The incidences of lymphoma of thymic origin were increased in exposed groups of males, but were significantly increased only in the 6,000 ppm group. The incidences of atypical hyperplasia of the thymus in 6,000 and 12,000 ppm males and in all exposed groups of females were significantly greater than those in the controls. The incidences of benign sex cord stromal tumors of the ovary in all exposed groups of females were significantly greater than in the controls. The incidences of hyperplasia of the ovary in 3,000 and 12,000 ppm females were significantly greater than in the controls. The incidences of germinal epithelial degeneration of the testis in all exposed groups of males were significantly greater than that in the controls. There were increased incidences of myelofibrosis of the bone marrow in 12,000 ppm males (0 ppm, 3/50; 3,000 ppm, 8/50; 6,000 ppm, 8/50; 12,000 ppm, 19/49) and an increased severity but not incidence of this lesion in exposed females. There were also increased incidences of pigmentation of minimal to mild severity in the bone marrow of 6,000 and 12,000 ppm males (0/50, 2/50, 5/50, 16/49) and females (2/50, 3/50, 11/50, 11/50). Also, the incidences of hematopoietic cell proliferation in the red pulp of the spleen (10/50, 22/50, 28/50, 21/49) in all exposed groups of males were significantly greater than that in the controls, and the severity of this lesion increased with increasing exposure concentration. The incidences of hepatocellular adenoma in all exposed groups of males and females and of hepatocellular adenoma or carcinoma (combined) in 6,000 and 12,000 ppm males and all exposed groups of females were significantly less than those in the controls, and these lesions occurred with significant negative trends. Multiple hepatocellular adenomas were observed more frequently in the control groups than in the exposed groups. The incidences of clear cell and eosinophilic foci in all exposed groups of males and of mixed cell foci in 12,000 ppm males were significantly less than those in the controls. The incidences of eosinophilic foci in exposed groups of females were significantly less than that in the controls. GENETIC TOXICOLOGY: Phenolphthalein, tested in two laboratories, was not mutagenic in any of four strains of Salmonella typhimurium with or without S9 metabolic activation enzymes, and no induction of sister chromatid exchanges was observed in cultured Chinese hamster ovary cells treated with phenolphthalein with or without S9. However, significant increases in chromosomal aberrations were observed after treatment of cultured Chinese hamster ovary cells with phenolphthalein in the presence of S9, and the frequencies of micronucleated erythrocytes were increased in peripheral blood samples from male and female mice administered phenolphthalein in feed for 13 weeks. CONCLUSIONS: Under the conditions of these 2-year feed studies, there was clear evidence of carcinogenic activity of phenolphthalein in male F344/N rats based on markedly increased incidences of benign pheochromocytomas of the adrenal medulla and of renal tubule adenomas and adenomas or carcinomas (combined). There was some evidence of carcinogenic activity of phenolphthalein in female F344/N rats based on the increased incidences of benign pheochromocytomas of the adrenal medulla in the 12,000 ppm group and of benign or malignant pheochromocytomas (combined) in the 12,000 and 25,000 ppm groups. There was clear evidence of carcinogenic activity of phenolphthalein in male B6C3F1 mice based on increased incidences of histiocytic sarcomas and of malignant lymphomas of thymic origin. There was clear evidence of carcinogenic activity of phenolphthalein in female B6C3F1 mice based on increased incidences of histiocytic sarcomas, malignant lymphomas of all types, lymphomas of thymic origin, and benign sex-cord stromal tumors of the ovary. Exposure of rats to phenolphthalein in feed for 2 years resulted in increased incidences of focal hyperplasia of the adrenal medulla in males and in increased incidences and/or severity of nephropathy of the kidney in males and females. Exposure of mice to phenolphthalein in feed for 2 years resulted in increased incidences of atypical hyperplasia of the thymus in males and females, degeneration of the germinal epithelium of the testis in males, and ovarian hyperplasia in females. Exposure of mice to phenolphthalein in feed for 2 years resulted in decreased incidences of hepatocellular neoplasms and nonneoplastic lesions in males and females. Synonyms: 3,3-Bis(4-hydroxyphenyl)-1(3H) isobenzofuranone; 3,3-bis( p-hydroxyphenyl)phthalide; a-p -hydroxyphenyl)-a- (4 oxo-2,5-cyclohexadien-1-ylidene)- o-toluic acid Trade names: Agoral®, Alophen®, Colax®, Correctol®, Dialose®, Doxidan®, Espotabs®, Evac-U-Gen®, Evac-U-Lax®, Ex-Lax®, Feen-A-Mint®, FemiLax®, Kondremul®, LaxCaps®, Lax-Pills®, Medilax®, Modane®, Phenolax®, Prulet® PMID- 12579200 TI - NTP Toxicology and Carcinogenesis Studies of Technical Grade Sodium Xylenesulfonate (CAS No. 1300-72-7) in F344/N Rats and B6C3F1 Mice (Dermal Studies). AB - Sodium xylenesulfonate is used as a hydrotrope, an organic compound that increases the ability of water to dissolve other molecules. Sodium xylenesulfonate is a component in a variety of widely used shampoos and liquid household detergents where it can constitute up to 10% of the total solution. Because of its widespread use, the potential for human exposure to sodium xylenesulfonate is great. Male and female F344/N rats and B6C3F1 mice were administered sodium xylenesulfonate in water or 50% ethanol dermally for 17 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, L5178Y mouse lymphoma cells, and cultured Chinese hamster ovary cells. 17-DAY STUDY IN RATS: Groups of five male and five female rats were administered 300 mL of 0, 5, 15, 44, 133, or 400 mg/mL sodium xylenesulfonate in distilled water by dermal application 5 days per week for 17 days. All rats survived to the end of the study. Final mean body weights and body weight gains of dosed rats were similar to those of the control groups. Dermal applications of 300 mL of 5, 15, 44, 133, and 400 mg/mL delivered average daily doses of approximately 10, 30, 90, 260, and 800 mg sodium xylenesulfonate/kg body weight to males and 13, 40, 120, 330, and 1,030 mg/kg to females. Clinical findings generally involved the skin of dosed animals and included tan or brown skin discoloration and crusty white deposits (presumed to be dried chemical) at the site of application. Neither of these observations were considered significant findings. The relative liver weights of 133 and 400 mg/mL male and female rats were significantly greater than those of the control groups, but the absolute liver weights were not increased and the biological significance of the relative differences in liver weight was unclear. In males and females, the few lesions observed grossly and microscopically were generally attributed to repeated clipping and were not considered related to chemical administration. 17-DAY STUDY IN MICE: Groups of five male and five female mice were administered 100 mL of 0, 5, 15, 44, 133, or 400 mg/mL sodium xylenesulfonate in distilled water by dermal application 5 days per week for 17 days. All mice survived to the end of the study. Final mean body weights and body weight gains of dosed mice were similar to those of the controls. Dermal applications of 5, 15, 44, 133, and 400 mg/mL delivered average daily doses of approximately, 20, 60, 190, 540, and 1,600 mg sodium xylenesulfonate/kg body weight to males and 26, 80, 220, 680, and 2,000 mg/kg to females. Clinical findings included crusty white deposits (presumed to be dried chemical) at the site of application in two 133 mg/mL males and in all 400 mg/mL males and females. The absolute and relative liver weights of 15 and 44 mg/mL males and 400 mg/mL males and females were significantly greater than those of the control groups, but the biological significance of these differences was unclear. The few skin lesions observed grossly and microscopically in males and females were generally attributed to repeated clipping and were not considered related to chemical administration. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were administered 300 mL of 0, 5, 15, 44, 133, or 400 mg/mL sodium xylenesulfonate in 50% ethanol by dermal application for 14 weeks. For special hematology and clinical pathology studies, additional groups of 10 male and 10 female rats were administered 0, 5, 15, 44, 133, or 400 mg/mL sodium xylenesulfonate in 50% ethanol by dermal application for 14 weeks. All rats survived to the end of the study. Final mean body weights and body weight gains of dosed male and female rats were similar to those of the control groups. Dermal applications of 5, 15, 44, 133, and 400 mg/mL delivered average daily doses of approximately 6, 20, 60, 170, and 500 mg sodium xylenesulfonate/kg body weight to males and 10, 30, 90, 260, and 800 mg/kg to females. The only notable clinical finding was brown discoloration of the skin at the site of application in dosed animals. Hemaation in dosed animals. Hematology and clinical chemistry parameters of dosed groups of males and females were significantly different from those of the controls in several instances, but these differences were sporadic and did not demonstrate a treatment relationship. The absolute and relative liver weights of males receiving 44, 133, or 400 mg/mL were significantly less than those of the control group, but the biological significance of these differences was unclear, and there were no treatment-related histopathologic effects in the liver. There were no significant differences in liver weights in female rats. Minimal hyperplasia of the epidermis at the site of application occurred in both male and female rats in the control group as well as most dosed groups. The incidence of epidermal hyperplasia in 400 mg/mL males was possibly chemical related. 14-Week Study in Mice: Groups of 10 male and 10 female mice were administered 100 mL of 0, 5, 15, 44, 133, or 400 mg/mL sodium xylenesulfonate in 50% ethanol by dermal application for 14 weeks. There were no chemical related deaths. The mean body weight gain of the 400 mg/mL males was significantly greater than that of the control group. Dermal applications of 5, 15, 44, 133, and 400 mg/mL delivered average daily doses of approximately 17, 40, 140, 440, and 1,300 mg sodium xylenesulfonate/kg body weight to males and 20, 60, 170, 530, and 1,630 mg/kg to females. There were no clinical findings related to sodium xylenesulfonate administration. Epidermal hyperplasia occurred in one 44 mg/mL female, two 133 mg/mL males, five 400 mg/mL males, and four 400 mg/kg females. Hyperplasia of the epidermis in 400 mg/mL males and females was probably related to chemical administration. Chronic inflammation of the skin occurred primarily in the control groups of males and females. These lesions consisted of mononuclear inflammatory cells in the dermis. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were dermally administered 0, 60, 120, or 240 mg sodium xylenesulfonate/kg body weight in 50% ethanol for 104 weeks. Survival, Body Weights, and Clinical Findings: Survival of dosed males and females was similar to that of the control groups. Mean body weights of dosed males and females were similar to those of the controls throughout the study. In male groups, there were no clinical findings considered treatment related. In females, clinical findings were limited to irritation at the site of application in one control female, four 120 mg/kg females, and two 240 mg/kg females. Pathology Findings: There were no neoplasms at any site (including the skin) that were considered treatment related.Low incidences of hyperplasia of the epidermis at the site of application occurred in males in the 60, 120, and 240 mg/kg groups. Low incidences of hyperplasia of the epidermis at the site of application also occurred in females in the 120 and 240 mg/kg groups, and they occurred with a significant positive trend. Low incidences of hyperplasia of the sebaceous gland occurred in control and 60 mg/kg males and in control, 120 mg/kg, and 240 mg/kg females. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were dermally administered 0, 182, 364, or 727 mg sodium xylenesulfonate/kg body weight in 50% ethanol for 104 to 105 weeks. Survival, Body Weights, and Clinical Findings: Survival of dosed males and females was similar to that of the control groups. Mean body weights of dosed males and females were generally similar to those of the controls throughout the study; however, the mean body weights of 727 mg/kg females were greater than those of the control group from week 85 to week 97. With the exception of irritation at the site of application in one 364 mg/kg female, there were no clinical findings related to sodium xylenesulfonate administration. Pathology Findings: There were no neoplasms at any site (including the skin) that were considered treatment related.Hyperplasia of the epidermis occurred in control,364 mg/kg, and 727 mg/kg males and in control and dosed females. In male mice, the incidences occurred with a significant positive trend. Focal ulceration occurred in one 727 mg/kg male and in one female in each dose group. In males and females from control and dosed groups, the incidences of hepatocellular adenoma, hepatocellular carcinoma, and hepato- cellular adenoma or carcinoma (combined) were generally higher than those expected by spontaneous occurrence. The incidences of hepatocellular neoplasms in some groups of males and females exceeded the NTP historical control range. Male mice had a pattern of nonneoplastic liver lesions along with silver stained positive helical organisms within the liver which suggests an infection with Helicobacter hepaticus. The findings in this study of sodium xylenesulfonate were not considered to have been significantly impacted by the infection with H. hepaticus or its associated hepatitis. GENETIC TOXICOLOGY: Sodium xylenesulfonate was not mutagenic in Salmonella typhimurium strain TA98, TA100, TA1535, or TA1537 with or without induced liver S9. Equivocal results were obtained in a mutation assay with mouse lymphoma cells in the presence of induced S9; no evidence of mutagenicity was noted without S9 in this assay. In cytogenetic tests with sodium xylenesulfonate in cultured Chinese hamster ovary cells, significant increases in sister chromatid exchanges were observed in the absence of S9 only, and no increases in chromosomal aberrations were observed with or without S9. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of sodium xylenesulfonate in male or female F344/N rats administered 60, 120, or 240 mg/kg or in male or female B6C3F1 mice administered 182, 364, or 727 mg/kg. Increased incidences of epidermal hyperplasia in female rats and male mice may have been related to exposure to sodium xylenesulfonate. Synonyms: Benzenesulfonic acid, dimethyl-, sodium salt; xylenesulfonic acid, sodium salt; sodium dimethylbenzenesulfonate; xylenesulfonic acid, sodium salt Trade names: Conco SXS; Cyclophil; SXS 30; Eletesol SX 30; Naxonate; Naxonate G; Richonate SXS; Stepanate SXS; Stepanate X; SXS 40; Ultrawet 40SX PMID- 12579201 TI - NTP Toxicology and Carcinogenesis Studies of Isobutyraldehyde (CAS No. 78-84-2) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Isobutyraldehyde, a branched alkyl aldehyde, is used as a chemical intermediate and flavoring agent. It was nominated by the National Cancer Institute for toxicity and carcinogenicity studies by the NTP. Reasons for nomination and selection of isobutyraldehyde for study included its high potential for human exposure as suggested by its high production volume, its use as a chemical intermediate and food flavoring agent, suspicion of carcinogenicity due to an increased incidence of cancer at an aldehyde manufacturing plant where workers were exposed to a variety of aldehydes, its structural relationship to formaldehyde (a nasal carcinogen in rats), and the lack of toxicity and carcinogenicity studies on isobutyraldehyde in animals. Although human exposure occurs orally, dermally, or via inhalation, the inhalation route of exposure was selected for these animal studies because of the instability of isobutyraldehyde in water and feed. Male and female F344/N rats and B6C3F1 mice were exposed to isobutyraldehyde (approximately 99% pure) by inhalation for 13 weeks or 2 years. Genetic toxicology studies were conducted in vitro in Salmonella typhimurium, L5178Y mouse lymphoma cells, and cultured Chinese hamster ovary cells; in vivo tests were conducted in Drosophila melanogaster germ cells and bone marrow cells of rats and mice. 13-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were exposed to 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm isobutyraldehyde by inhalation, 6 hours per day, 5 days a week, for 13 weeks. All rats exposed to 8,000 ppm died before the end of the study. Three male rats and six female rats in the 4,000 ppm groups and one female in the 500 ppm group died before the end of the study. The final mean body weight of male rats in the 4,000 ppm group and the body weight gains of 4,000 ppm males and females were significantly less than those of the chamber controls. Clinical findings in rats exposed to 4,000 or 8,000 ppm included abnormal respiratory sounds, decreased activity, nasal discharge, prostration, and slowed respiration. A minimal mature neutrophilia, evidenced by increased segmented neutrophil numbers, occurred in exposed groups of male and female rats. Exposure to isobutyraldehyde resulted in minimal increases in alanine aminotransferase activity in all groups of male and female rats. Spermatozoal motility in 500 and 1,000 ppm males was significantly reduced and females exposed to 4,000 ppm differed significantly from the chamber control females in the relative time spent in the estrous stages. No gross lesions were observed at necropsy that could be associated with isobutyraldehyde exposure. In the 8,000 ppm groups, severe necrosis of the epithelium, and occasionally of the entire mucosa, of the nasal turbinates accompanied by an acute inflammatory reaction was observed. Increased incidences of squamous metaplasia and mild acute inflammation occurred in male and female rats exposed to 4,000 ppm. Minimal to mild degeneration of the olfactory epithelium was observed in all male rats in the 2,000 and 4,000 ppm groups. Male rats exposed to 4,000 or 8,000 ppm and females exposed to 4,000 ppm had mild osteodystrophy of the turbinate bone. The incidences of necrosis/degeneration of the larynx and trachea were increased in male rats in the 8,000 ppm group. The incidences of mild to moderate lymphoid depletion of the spleen and thymus and lymphoid necrosis of the thymus were significantly increased in male and female rats exposed to 8,000 ppm. 13-WEEK STUDY IN MICE: Ten male and 10 female B6C3F1 mice were exposed to 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm isobutyraldehyde by inhalation, 6 hours per day, 5 days per week, for 13 weeks. One male in the chamber control group, one male in the 1,000 ppm group, nine males and all females in the 4,000 ppm groups, and all males and females in the 8,000 ppm groups died before the end of the study. The final mean body weight and body weight gain of female mice in the 1,000 ppm group were significantly less than those of the chamber controls. Clinical findal findings included decreased activity, tremors, prostration, and slower and labored respiration. The absolute and relative kidney weights of males in the 1,000 and 2,000 ppm groups were significantly increased. There were no gross lesions observed at necropsy that could be associated with isobutyraldehyde exposure. Histopathologically, the nasal cavity and lymphopoietic tissues were considered target organs, with changes similar, but not identical, to those observed in rats. Increased incidences of nonneoplastic lesions of the nasal cavity were observed in male and female mice exposed to 1,000 ppm or greater. These lesions included necrosis, inflammation, hyperplasia, and squamous metaplasia of the epithelium; serous and suppurative exudate within the nasal passages; olfactory epithelial degeneration; and osteodystrophy of the turbinate bone. Mild to moderate lymphoid depletion and/or lymphoid necrosis were observed in the thymus of male and female mice exposed to 8,000 ppm. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to 0, 500, 1,000, or 2,000 ppm isobutyraldehyde by inhalation, 6 hours per day, 5 days per week, for 105 weeks Survival, Body Weights, and Clinical Findings No differences in survival rates between exposed and chamber control rats were found. The mean body weights of male and female rats were generally similar to those of the chamber controls throughout the study. Pathology Findings No increase in neoplasm incidences that could be attributed to exposure to isobutyraldehyde was observed in male or female rats. Nonneoplastic lesions related to isobutyraldehyde exposure were limited to the nose and consisted of squamous metaplasia of the respiratory epithelium, degeneration of the olfactory epithelium, and suppurative inflammation. Incidences of minimal to mild squamous metaplasia in 1,000 and 2,000 ppm males and females and in 500 ppm females were significantly greater than those in the chamber controls. Another lesion associated with isobutyraldehyde exposure was minimal to mild degeneration of the olfactory epithelium in 2,000 ppm males and females. The incidences of suppurative inflammation (rhinitis) in male and female rats exposed to 2,000 ppm were increased compared to the chamber controls. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were exposed to 0, 500, 1,000, or 2,000 ppm isobutyraldehyde by inhalation, 6 hours per day, 5 days per week, for 105 weeks. Survival, Body Weights, and Clinical Findings There was an exposure-related decrease in survival of male mice, and the survival of males exposed to 2,000 ppm was marginally lower than that of the chamber controls. The mean body weights of female mice exposed to 1,000 or 2,000 ppm were lower than those of the chamber controls during the second year of the study. Pathology Findings No neoplasms that could be attributed to iso butyraldehyde exposure were observed in mice. Non neoplastic lesions related to isobutyraldehyde exposure were limited to the nose. The incidences of olfactory epithelial degeneration in 1,000 and 2,000 ppm males and females were significantly greater than in the chamber controls. GENETIC TOXICOLOGY: Isobutyraldehyde is mutagenic in vitro and in vivo, with the strongest responses observed in mammalian cell assays that measured chromosomal damage. Results of an initial mutagenicity test in S. typhimurium were negative; a second test, con ducted with different strains and varying concentrations of induced S9 activation enzymes, gave equivocal results. Strongly positive responses were obtained in the mouse lymphoma assay for mutation induction in L5178Y cells without S9 and in cytogenetic tests for induction of sister chromatid exchanges and chromosomal aberrations in cultured Chinese hamster ovary cells. Sister chromatid exchanges were significantly increased with and without S9, but induction of chromosomal aberrations was noted unequivocally only in the absence of S9. No induction of sex-linked recessive lethal mutations was observed in germ cells of male D. melanogaster administered isobutyraldehyde by feeding or by injection. In vivo, isobutyraldehyde was demonstrated to induce chromosomal aberrations in bone marrow cells of male mice, but no increases in micronuclei were observed in bone marrow cells of mice or rats after administration of isobutyraldehyde. All these in vivo cytogenetic studies used doses that reached lethality CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was no evidence of carcinogenic activity of isobutyraldehyde in male or female F344/N rats or male or female B6C3F1 mice exposed to 500, 1,000, or 2,000 ppm isobutyraldehyde. In male and female rats, exposure to isobutyraldehyde induced squamous metaplasia and suppurative inflammation of the nasal respiratory epithelium and degeneration of the nasal olfactory epithelium. In male and female mice, exposure to isobutyraldehyde caused degeneration of the nasal olfactory epithelium. Synonyms: Dimethylacetaldehyde; 2-formylpropane; isobutanal; isobutylcarboxaldehyde; isobutyral; isobutyric aldehyde; isobutyrylaldehyde; isopropylformaldehyde; 2-methylpropanal; 2-methyl-1-propanal; a methylpropionaldehyde; 2-methylpropionaldehyde; valine aldehyde PMID- 12579202 TI - NTP Toxicology and Carcinogenesis Studies of Cobalt Sulfate Heptahydrate (CAS No. 10026-24-1) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Cobalt sulfate is used in the electroplating and electro chemical industries. It is also used as a coloring agent for ceramics and as a drying agent in inks, paints, varnishes, and linoleum. Cobalt sulfate may be added to animal feed as a mineral supplement and has been used as a top dressing on pasture lands. Cobalt sulfate was nominated by the National Cancer Institute for study based on a lack of information on the toxicity of soluble salts. Male and female F344/N rats and B6C3F1 mice were exposed to cobalt sulfate heptahydrate (approximately 99% pure) by inhalation for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium. The results of prechronic inhalation toxicity studies were reported previously (Bucher et al., 1990; NTP, 1991). 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed to aerosols containing 0, 0.3, 1.0, or 3.0 mg/m3 cobalt sulfate heptahydrate 6 hours per day, 5 days per week, for 105 weeks. Survival and Body Weights Survival of exposed males and females was similar to that of the chamber controls. Mean body weights of exposed male and female rats were similar to those of the chamber controls throughout the study. Pathology Findings The incidences and severities of proteinosis, alveolar epithelial metaplasia, granulomatous alveolar inflammation, and interstitial fibrosis were markedly greater in all exposed groups of male and female rats than in the chamber controls. The incidences of alveolar epithelial hyperplasia in all groups of exposed males and in females exposed to 3.0 mg/m3 were significantly greater than those in the chamber control groups, as were the incidences of squamous metaplasia in 1.0 mg/m3 females and atypical alveolar epithelial hyperplasia in 3.0 mg/m3 females. In 3.0 mg/m3 males, the combined incidence of alveolar/ bronchiolar neoplasms (adenoma and/or carcinoma) was significantly greater than in the chamber controls. In female rats exposed to 1.0 or 3.0 mg/m3, the incidences of alveolar/bronchiolar neoplasms were significantly greater than those in the chamber control group and exceeded the NTP historical control ranges. A squamous cell carcinoma was observed in one 1.0 mg/m3 and one 3.0 mg/m3 female. The incidences of benign, complex, or malignant pheochromocytoma (combined) in 1.0 mg/m3 males and in 3.0 mg/m3 females were significantly greater than those in the chamber controls and exceeded the historical control ranges. Hyperplasia of the lateral wall of the nose, atrophy of the olfactory epithelium, and squamous metaplasia of the epiglottis were observed in all exposed groups of males and females, and the severities of these lesions increased with increasing exposure concentration. The incidences of squamous metaplasia of the lateral wall of the nose and metaplasia of the olfactory epithelium were increased in 3.0 mg/m3 males and females. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed to aerosols containing 0, 0.3, 1.0, or 3.0 mg/m3 cobalt sulfate heptahydrate 6 hours per day, 5 days per week, for 105 weeks. Survival and Body Weights Survival of exposed males and females was similar to that of the chamber controls. Mean body weights of 3.0 mg/m3 male mice were less than those of the chamber controls from week 96 until the end of the study. The mean body weights of all exposed groups of female mice were generally greater than those of the chamber controls from week 20 until the end of the study. Pathology Findings The incidences of diffuse histiocytic cell infiltration in 3.0 mg/m3 males and of focal histiocytic cell infil tration in 3.0 mg/m3 females were significantly greater than those in the chamber controls. The incidences of alveolar/bronchiolar neoplasms in 3.0 mg/m3 males and females were significantly greater than those in the chamber control groups. The combined incidences of alveolar/bronchiolar adenoma or carcinoma and the incidences of alveolar/bronchiolar carcinoma in 3.0 mg/m3 males and females and the incidence of alveolar/bronchiolar adenoma in 3.0 mg/m3 females exceeded the NTP historical crical control ranges for inhalation studies. The incidences of atrophy of the olfactory epithelium in 1.0 and 3.0 mg/m3 males and females and hyper plasia of the olfactory epithelium in 3.0 mg/m3 males and females were significantly greater than in the chamber controls. Squamous metaplasia of the larynx was observed in all exposed groups of males and females. Male mice had a pattern of nonneoplastic liver lesions along with silver-staining helical organisms within the liver, characteristic of an infection with Helico bacter hepaticus. In NTP studies with H. hepaticus- associated hepatitis, increased incidences of hemangiosarcoma were seen in the liver of male mice. In this study of cobalt sulfate heptahydrate, incidences of hemangiosarcoma were increased in exposed groups of male mice. Because of the above association, interpretation of the increased incidences of hemangiosarcoma in the livers of male mice was confounded. Incidences of lesions at other sites in this study of cobalt sulfate heptahydrate were not considered to have been significantly impacted by the infection with H. hepaticus or its associated hepatitis. GENETIC TOXICOLOGY: Cobalt sulfate heptahydrate was mutagenic in S. typhimurium strain TA100 with and without liver S9 metabolic activation enzymes; no mutagenic activity was detected in strain TA98 or TA1535, with or without S9. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was some evidence of carcinogenic activity of cobalt sulfate heptahydrate in male F344/N rats based on increased incidences of alveolar/bronchiolar neoplasms. Marginal increases in incidences of pheochromocytomas of the adrenal medulla may have been related to exposure to cobalt sulfate heptahydrate. There was clear evidence of carcinogenic activity in female F344/N rats based on increased incidences of alveolar/bronchiolar neo plasms and pheochromocytomas of the adrenal medulla in groups exposed to cobalt sulfate heptahydrate. There was clear evidence of carcinogenic activity of cobalt sulfate heptahydrate in male and female B6C3F1 mice based on increased incidences of alveolar/bronchiolar neoplasms. Exposure to cobalt sulfate heptahydrate caused a spectrum of inflammatory, fibrotic, and proliferative lesions in the respiratory tract of male and female rats and mice. Synonyms: Bieberite; cobalt(II) sulfate (1:1) heptahydrate; cobalt monosulfate heptahydrate; cobalt(II) sulphate heptahydrate; sulfuric acid, cobalt(2+) salt (1:1) heptahydrate PMID- 12579203 TI - NTP Toxicology and Carcinogenesis Studies of Pyridine (CAS No. 110-86-1) in F344/N Rats, Wistar Rats, and B6C3F1 Mice (Drinking Water Studies). AB - Pyridine is used as a denaturant in alcohol and anti freeze mixtures, as a solvent for paint, rubber, and polycarbonate resins, and as an intermediate in the manufacture of insecticides, herbicides, and fungicides. It is used in the production of piperidine, an intermediate in the manufacture of rubber and mepiquat chloride, and as an intermediate and solvent in the preparation of vitamins and drugs, dyes, textile water repellants, and flavoring agents in food. Pyridine was nominated for study because of its large production volume and its use in a variety of food, medical, and industrial products. Male and female F344/N rats, male Wistar rats, and male and female B6C3F1 mice were exposed to pyridine (approximately 99% pure) in drinking water for 13 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, L5178Y mouse lymphoma cells, cultured Chinese hamster ovary cells, Drosophila melanogaster, and mouse bone marrow cells. 13-WEEK STUDY IN F344/N RATS: Groups of 10 male and 10 female F344/N rats were exposed to pyridine in drinking water at concentrations of 0, 50, 100, 250, 500, or 1,000 ppm (equivalent to average daily doses of 5, 10, 25, 55, or 90 mg pyridine/kg body weight). Two females exposed to 1,000 ppm died during week 1. Final mean body weights of 1,000 ppm males and females and 500 ppm females were significantly less than controls. Water consumption by female rats exposed to 1,000 ppm was less than that by controls. At study termination, evidence of anemia persisted in the 500 and 1,000 ppm males and all exposed groups of females. There was evidence of hepatocellular injury and/or altered hepatic function demonstrated by increased serum alanine aminotransferase and sorbitol dehydrogenase activities and bile acid concentrations in 500 and 1,000 ppm rats. The estrous cycle length of 1,000 ppm females was significantly longer than that of the controls. Liver weights of males and females exposed to 250 ppm or greater were significantly greater than controls. In the liver, the incidences of centrilobular degeneration, hypertrophy, chronic inflammation, and pigmentation were generally increased in 500 and 1,000 ppm males and females relative to controls. In the kidney, the incidences of granular casts and hyaline degeneration (hyaline droplets) were significantly increased in 1,000 ppm males and slightly increased in 500 ppm males; these lesions are consistent with 2u-globulin nephropathy. Additionally, there were increased incidences and/or severities of protein casts, chronic inflammation, mineralization, and regeneration primarily in 500 and 1,000 ppm males. 13-WEEK STUDY IN MALE WISTAR RATS: Groups of 10 male Wistar rats were exposed to pyridine in drinking water at concentrations of 0, 50, 100, 250, 500, or 1,000 ppm (equivalent to average daily doses of 5, 10, 30, 60, or 100 mg/kg). One male rat exposed to 500 ppm died during week 1. Final mean body weights of rats exposed to 250, 500, or 1,000 ppm were significantly less than those of the controls. Water consumption by rats exposed to 1,000 ppm was lower than that by controls. There was evidence of hepatocellular injury and/or altered hepatic function in the 500 and 1,000 ppm groups, similar to that observed in the 13-week study in F344/N rats. Incidences of centrilobular degeneration, hypertrophy, chronic inflammation, and pigmentation in the liver of rats exposed to 500 or 1,000 ppm were significantly increased relative to controls. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were exposed to pyridine in drinking water at concentrations of 0, 50, 100, 250, 500, or 1,000 ppm (equivalent to average daily doses of 10, 20, 50, 85, or 160 mg/kg for males and 10, 20, 60, 100, or 190 mg/kg for females). One female mouse exposed to 250 ppm died during week 2. Final mean body weights of female mice exposed to 1,000 ppm were significantly less than those of controls. Water consumption by exposed female mice was lower than that by controls at week 1 but generally slightly higher than controls at week 13. Sperm motirm motility in exposed male mice was significantly decreased relative to controls. Liver weights were significantly increased relative to controls in males exposed to 100 ppm or greater and in 250 and 500 ppm females. No chemical-related lesions were observed in male or female mice. 2-YEAR STUDY IN F344/N RATS: Groups of 50 male and 50 female F344/N rats were exposed to pyridine in drinking water at concentrations of 0, 100, 200, or 400 ppm (equivalent to average daily doses of 7, 14, or 33 mg/kg) for 104 (males) or 105 (females) weeks. Survival, Body Weights, and Water Consumption Survival of exposed males and females was similar to that of controls. Mean body weights of 400 ppm males and females were generally less than those of the controls throughout the study, and those of 200 ppm males and females were less during the second year of the study. Water consumption by males and females exposed to 200 or 400 ppm was generally greater than that by controls. Pathology Findings Incidences of renal tubule adenoma and renal tubule adenoma or carcinoma (combined) in male rats exposed to 400 ppm were significantly increased compared to controls and exceeded the historical control ranges. The findings from an extended evaluation (step section) of the kidneys did not reveal additional carcinomas, but additional adenomas were observed in each group of males. In the standard evaluation, an increased incidence of renal tubule hyperplasia was observed in 400 ppm males compared to controls. Incidences of mononuclear cell leukemia in female rats were significantly increased in the 200 and 400 ppm groups, and the incidence in the 400 ppm group exceeded the historical control range. Exposure concentration-related nonneoplastic liver lesions were observed in males and females, and the incidences were generally increased in groups exposed to 400 ppm. These included centrilobular cytomegaly, cytoplasmic vacuolization, periportal fibrosis, fibrosis, centrilobular degeneration and necrosis, and pigmentation. Bile duct hyperplasia occurred more often in exposed females than in controls. 2-YEAR STUDY IN MALE WISTAR RATS: Groups of 50 male Wistar rats were exposed to pyridine in drinking water at concentrations of 0, 100, 200, or 400 ppm (equivalent to average daily doses of 8, 17, or 36 mg/kg) for 104 weeks. Survival, Body Weights, and Water Consumption Survival of rats exposed to 200 or 400 ppm was significantly less than that of the controls. Mean body weights of rats exposed to 100, 200, or 400 ppm were significantly less than controls. Water consumption was similar by control and exposed rats. Pathology Findings The incidence of testicular interstitial cell adenoma in rats exposed to 400 ppm was significantly increased compared to controls. Incidences of interstitial cell hyperplasia were observed in control and exposed groups and were slightly, but not significantly, increased in rats exposed to 200 or 400 ppm. Severity of nephropathy was marked in all groups, and additional evidence of kidney disease, including mineralization in the glandular stomach, parathyroid gland hyperplasia, and fibrous osteodystrophy, was observed in 100 and 200 ppm rats. The incidences of hepatic centrilobular degeneration and necrosis, fibrosis, periportal fibrosis, and/or pigmentation were increased in one or more exposed groups. 2-YEAR STUDY IN MICE: Groups of 50 male B6C3F1 mice were exposed to pyridine in drinking water at concentrations of 0, 250, 500, or 1,000 ppm (equivalent to average daily doses of 35, 65, or 110 mg/kg) for 104 weeks, and groups of 50 female B6C3F1 mice were exposed to pyridine in drinking water at concentrations of 0, 125, 250, or 500 ppm (equivalent to average daily doses of 15, 35, or 70 mg/kg) for 105 weeks. Survival, Body Weights, and Water Consumption Survival of exposed males and females was similar to that of the controls. Mean body weights of 250 and 500 ppm females were less than controls. Water consumption by males exposed to 250 or 500 ppm was generally greater than that by controls during the last year of the study; male mice exposed to 1,000 ppm consumed less water than controls throughout the study. Water consumption by exposed females was generally lower than that by controls during the first year of the study, but greater than controls during the second year. Pathology Findings Hepatocellular neoplasms, including hepatoblastomas, in exposed male and female mice were clearly related to pyridine exposure. Additionally, many mice had multiple hepatocellular neoplasms. The incidences of hepatocellular neoplasms in exposed males and females generally exceeded the historical control ranges for drinking water studies. Neoplasms from control mice, 1,000 ppm males, and 500 ppm females were negative when stained for p53 protein. GENETIC TOXICOLOGY: Pyridine was not mutagenic in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 or in L5178Y mouse lymphoma cells, with or without S9 metabolic activation, and it did not induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells, with or without S9. Pyridine was tested for induction of sex-linked recessive lethal mutations in adult male Drosophila melanogaster, and mixed results were obtained. In one experiment, administration by injection gave negative results, but feeding produced an equivocal response. A second experiment generated negative results by injection and feeding. A third experiment showed significant increases in sex-linked recessive lethal mutations in flies treated with pyridine by injection but not by feeding. Overall, results of the sex-linked recessive lethal mutations test in Drosophila melanogaster were considered negative by feeding and equivocal by injection. Results of a single reciprocal translocation test in male Drosophila melanogaster were negative. No induction of chromosomal aberrations or micronuclei was noted in bone marrow cells of male mice administered pyridine via intraperitoneal injection. CONCLUSIONS: Under the conditions of these 2-year drinking water studies, there was some evidence of carcinogenic activity of pyridine in male F344/N rats based on increased incidences of renal tubule neoplasms. There was equivocal evidence of carcinogenic activity of pyridine in female F344/N rats based on increased incidences of mononuclear cell leukemia. There was equivocal evidence of carcinogenic activity in male Wistar rats based on an increased incidence of interstitial cell adenoma of the testis. There was clear evidence of carcinogenic activity of pyridine in male and female B6C3F1 mice based on increased incidences of malignant hepatocellular neoplasms. In F344/N rats, exposure to pyridine resulted in increased incidences of centrilobular cytomegaly and degeneration, cytoplasmic vacuolization, and pigmentation in the liver of males and females; periportal fibrosis, fibrosis, and centrilobular necrosis in the liver of males; and bile duct hyperplasia in females. In male Wistar rats, pyridine exposure resulted in increased incidences of centrilobular degeneration and necrosis, fibrosis, periportal fibrosis, and pigmentation in the liver, and, secondary to kidney disease, mineralization in the glandular stomach and parathyroid gland hyperplasia. Synonyms: Azabenzene, azine PMID- 12579204 TI - NTP Toxicology and Carcinogenesis Studies of AZT (CAS No. 30516-87-1) and AZT/alpha-Interferon A/D B6C3F1 Mice (Gavage Studies). AB - 3'-Azido-3'-deoxythymidine (AZT) is the most widely used and evaluated chemotherapeutic agent for the treatment of persons with acquired immune deficiency syndrome (AIDS) and persons seropositive for human immunodeficiency virus (HIV). The National Cancer Institute nominated AZT for toxicity and carcinogenicity studies because of the impending large-scale use of AZT in the treatment of adult patients with AIDS or AIDS-related complex. alpha-Interferon A/D, which displays antiviral activity in mice, is a hybrid molecule composed of the N-terminal portion of human alpha-interferon A and the C-terminal portion of human alpha-interferon D. AZT and alpha-interferon A/D combination studies were conducted because in vitro studies of AZT and alpha-interferon have demonstrated that the combination is more effective in blocking HIV infection than either agent alone. Male and female B6C3F1 mice received AZT (approximately 98% pure) in 0.5% aqueous methylcellulose by gavage for 14 weeks or 2 years. In addition, male and female B6C3F1 mice received alpha-interferon A or alpha-interferon A/D by subcutaneous injection for 2 years, and male and female B6C3F1 mice received AZT in 0.5%% aqueous methylcellulose by gavage in combination with alpha-interferon A/D by subcutaneous injection for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, mouse bone marrow erythrocytes, and mouse peripheral blood erythrocytes. 14-WEEK AZT STUDY: Groups of 10 male and 10 female mice received AZT in 0.5% methylcellulose by gavage at doses of 0, 50, 100, 200, 800, or 2,000 mg/kg daily for 14 weeks. Additional groups of 10 male and 10 female mice received AZT in 0.5% methylcellulose by gavage at doses of 0, 100, 800, or 2,000 mg/kg daily for 14 weeks and then were held without treatment for an additional 4 weeks before necropsy. One female receiving 100 mg/kg and two females receiving 200 mg/kg died during week 1 as a result of gavage trauma; one female receiving 2,000 mg/kg also died prior to the end of the 14-week dosing period. One female receiving 2,000 mg/kg in the recovery study also died from gavage trauma during week 1. The final mean body weights of dosed mice were similar to those of the vehicle control groups at the end of the dosing period and at the end of the recovery period. Female mice receiving 200, 800, or 2,000 mg/kg gained less weight than the vehicle controls during the 14-week dosing period. Exposure to AZT was toxic to the bone marrow, resulting in significant changes in the peripheral blood (decreased hematocrit values, erythrocyte counts, and hemoglobin concentrations, and increased mean cell volume and mean cell hemoglobin) and bone marrow (erythroid hypoplasia) characteristic of a dose- and time-dependent, minimal to moderate, poorly regenerative macrocytic anemia. At the end of the 4-week recovery period, the hematology parameters had returned to normal, indicating that the hematotoxicity was reversible. 2-YEAR STUDIES: AZT Groups of 95 male and 95 female mice received AZT in 0.5% methylcellulose by gavage at daily doses of 0, 30, 60, or 120 mg/kg body weight, administered as two equal doses at least 6 hours apart, 5 days per week for 105 weeks. Each group of 95 animals was composed of a core group of 50 animals for evaluation of carcinogenic response, a group of 30 animals for evaluation of hematology and bone marrow cellularity, and a group of 15 animals from which blood was drawn for determination of plasma AZT concentrations at week 54. alpha-Interferon A/D and AZT/alpha-Interferon A/D Studies Groups of 80 male and 80 female mice received AZT in 0.5% aqueous methylcellulose by gavage at daily doses of 0, 30, 60, or 120 mg/kg body weight, given in two equal doses, 5 days per week for 105 weeks. Those groups receiving AZT also received sub-cutaneous injections of 500 or 5,000 U alpha-interferon A/D three times per week for 105 weeks. Additional groups of 80 male and 80 female mice received subcutaneous injections of the vehicle, 500 U alpha-interferon A/D, 5,000 Uutaneous injections of the vehicle, 500 U α-interferon A/D, 5,000 U α-interferon A/D, or 5,000 U α-interferon A, three times per week for 105 weeks. Each group of 80 animals was composed of a core group of 50 animals for evaluation of carcinogenic response and a group of 30 animals for evaluation of hematology and bone marrow cellularity. Because of the large number of animals involved, the 2-year studies were started in four phases and, for clarity, are presented as follows: the AZT study, the α-interferon A/D study, the AZT/500 U α-interferon A/D study, and the AZT/5,000 U α-interferon A/D study. Design of the 2-year AZT, AZT/α-Interferon A/D, and α Interferon A/D Studies AZT Dose AZT Study AZT/500 U α-Interferon A/D Study AZT/5,000 U α-Interferon A/D Study 500 or 5,000 U α-Interferon A/D or 5,000 U α-Interferon A Study Vehicle Control 95 male and 95 female micea 80 male and 80 female miceb 80 male and 80 female miceb 80 male and 80 female miceb 30 mg/kg AZT 95 male and 95 female mice 80 male and 80 female mice 80 male and 80 female mice none 60 mg/kg AZT 95 male and 95 female mice 80 male and 80 female mice 80 male and 80 female mice none 120 mg/kg AZT 95 male and 95 female mice 80 male and 80 female mice 80 male and 80 female mice none aFor the AZT study, there were 95 male and 95 female mice; these were divided into 50 males and 50 females in the core groups, 30 males and 30 females in the clinical pathology groups (hematology and bone marrow analyses only), and 15 males and 15 females for plasma AZT concentration determinations. bFor the α-interferon A/D study and the AZT/α-interferon A/D studies, there were 80 male and 80 female mice for each study; these were divided into 50 males and 50 females in the core groups and 30 males and 30 females in the clinical pathology groups (hematology and bone marrow analyses only). Survival and Body Weights Survival and mean body weights of mice exposed to AZT, α-interferon A, α-interferon A/D, or AZT plus α-interferon A/D were generally similar to those of the vehicle control groups. Hematology and Bone Marrow Analyses All groups of male and female mice receiving AZT exhibited changes in peripheral blood and bone marrow characteristic of a dose- and time-dependent, minimal to mild, macrocytic, nonresponsive anemia. In females, these changes were evident throughout the study. In males, the macrocytic anemia had resolved by week 80 in the 30 mg/kg group; at study termination erythrocyte macrocytosis was present only in males receiving 60 or 120 mg/kg AZT or AZT plus α-interferon A/D. There were no treatment-related alterations in hematology or bone marrow parameters in groups that received only α-interferon A or A/D. Pathology Findings Incidences of squamous cell carcinoma and squamous cell papilloma or carcinoma (combined) of the vagina occurred with a positive trend and were significantly increased in groups of female mice receiving 60 or 120 mg/kg AZT alone or in combination with α-interferon A/D. Epithelial hyperplasia was observed in all dosed groups of females, and the incidence was significantly increased in the 120 mg/kg AZT group. Three renal tubule adenomas and one renal tubule carcinoma were observed in male mice receiving 120 mg/kg AZT; the combined incidence in this group exceeded the range in historical controls. A renal tubule adenoma was observed in one male receiving 60 mg AZT/kg and 500 U α-interferon A/D; how ever, none were observed in other groups. Evaluation of step sections revealed a few more renal tubule hyperplasias but no additional neoplasms. The incidence of harderian gland adenoma was increased in male mice receiving 120 mg/kg AZT and exceeded the range in historical controls. Harderian gland neoplasms were observed in other groups but did not follow a treatment-related pattern. Overall Incidences of Vaginal Neoplasms and Hyperplasia of the Vaginal Epithelium in Female Mice in the 2-Year Gavage Studies of AZT and AZT/α-Interferon A/Da Vehicle Control 30 mg AZT/kg 60 mg AZT/kg 120 mg AZT/kg AZT alone 2/197 (1%)b 1/197 0/49 (0%) 3/49 5/45 (11%%) 4/45 11/49 (22%%) 11/49 500 U α Interferon A/D 0/49 (0%%) 0/49 0/44 (0%) 4/44 5/48 (10%) 8/48 6/48 (13%) 12/48 5,000 U α-Interferon A/D 1/50 (2%) 1/50 1/48 (2%) 4/48 5/48 (10%) 8/48 4/50 (8%) 15/50 aData are presented as number of vaginal neoplasms/number of animals microscopically examined (first line) and number of vaginal hyperplasias/number of animals microscopically examined (second line) bCombined incidences of controls from the AZT alone study and the AZT/α-interferon A/D studies; incidences in the vehicle control group from the AZT alone study are 0/50 (0%%) (neoplasms) and 0/50 (hyperplasia) Overall Incidence of Harderian Gland Neoplasms in Male Mice in the 2-Year Gavage Studies of AZT and AZT/α-Interferon A/Da Vehicle Control 30 mg AZT/kg 60 mg AZT/kg 120 mg AZT/kg AZT alone 13/200 (6%%)b 5/50 (10%%) 2/50 (4%) 10/50 (20%%) 500 U α-Interferon A/D 3/50 (6%) 3/50 (6%) 1/50 (2%%) 4/50 (8%%) 5,000 U α-Interferon A/D 3/50 (6%) 9/50 (18%%) 4/50 (8%%) 4/50 (8%) aData are presented as number of harderian gland neoplasms/number of animals necropsied bCombined incidences of controls from the AZT alone study and the AZT/α-interferon A/D studies; incidence in the vehicle control group from the AZT alone study is 3/50 (6%) Male mice had a pattern of nonneoplastic liver lesions along with silver-staining helical organisms within the liver consistent with an infection with Helicobacter hepaticus. An organism compatible with H. hepaticus was confirmed by polymerase chain reaction restriction fragment length polymorphism-based assays. Detection of dose-related differences in neoplasm incidences in these studies was not considered to have been significantly impacted by the infection with H. hepaticus or its associated hepatitis. GENETIC TOXICOLOGY: AZT is mutagenic in vitro and in vivo. It induced gene mutations in Salmonella typhimurium strain TA102, with and without S9; no increases in mutations were noted in the other tested strains of S. typhimurium. AZT induced sister chromatid exchanges, but not chromosomal aberrations, in cultured Chinese hamster ovary cells, with and without S9. In vivo studies with male mice administered AZT by gavage showed highly significant increases in micronucleated erythrocytes in bone marrow and peripheral blood after exposure periods that ranged from 72 hours to 14 weeks. CONCLUSIONS: Under the conditions of these 2-year gavage studies there was equivocal evidence of carcinogenic activity of AZT in male mice based on increased incidences of renal tubule and harderian gland neoplasms in groups receiving AZT alone. There was clear evidence of carcinogenic activity of AZT in female mice based on increased incidences of squamous cell neoplasms of the vagina in groups that received AZT alone or in combination with α-interferon A/D. Hematotoxicity occurred in all groups that received AZT. Treatment with AZT alone and AZT in combination with α interferon A/D resulted in increased incidences of epithelial hyperplasia of the vagina in all dosed groups of females. Synonyms: AZT; 3'-azido-2',3' dideoxythymidine; azidodeoxythymidine; azidothymidine; 3'-azidothymidine; 3' deoxy-3'-azidothymidine; 3'-deoxy-(8CI) (9CI); BW A509U; Compound S; ZDV; zidovudine Trade name: Retrovir® PMID- 12579205 TI - NTP Toxicology and Carcinogenesis Studies of Oxazepam (CAS No. 604-75-1) in F344/N Rats (Feed Studies). AB - Oxazepam and related benzodiazepine drugs are used in the treatment of anxiety. All benzodiazepines currently in use share a number of effects, including sedation, hypnosis, decreased anxiety, muscle relaxation, amnesia, and anticonvulsant activity. Oxazepam and four other benzodiazepines (chlordiazepoxide, chlorazepate, diazepam, and flurazepam) were nominated for study by the Food and Drug Administration (FDA) and by the NIEHS based on their widespread use, use by pregnant women, and the lack of adequate rodent carcinogenicity studies. Oxazepam was evaluated in 14-week and 2-year studies by the NTP, and Technical Report No. 443 contains the results of the studies performed with the Swiss-Webster and B6C3F1 strains of mice. Studies with rats were not initiated at the same time as the mouse studies because adequate carcinogenicity studies of oxazepam with the Sprague-Dawley rat strain had been submitted to the FDA. Subsequently, because of the marked neoplastic responses found in the two mouse strains, the NTP initiated 2-year studies of oxazepam with the F344/N rat. Groups of male and female F344/N rats were exposed to oxazepam (greater than 99% pure) in feed for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and cultured Chinese hamster ovary cells, and mouse peripheral blood samples were analyzed for the frequency of micronucleated normochromatic erythrocytes. 2-YEAR STUDY: Groups of 50 male and 50 female F344/N rats were fed diets containing 0, 625, 2,500, or 5,000 ppm oxazepam for up to 105 weeks. A stop-exposure group of 50 males and 50 females received 10,000 ppm oxazepam in feed for 26 weeks, after which animals received undosed feed for the remainder of the 2-year study. The continuous-exposure concentrations resulted in average daily doses of 25, 100, or 250 mg oxazepam/kg body weight to males and 25, 110, or 220 mg/kg to females. Stop- exposure males and females received an average daily dose of 630 mg/kg during the exposure period. Survival, Body Weights, and Clinical Findings: All 5,000 ppm continuous-exposure and 10,000 ppm stop-exposure males died before the end of the study. Survival of 2,500 ppm continuous-exposure males and females was significantly less than that of the controls. The mean body weight gains of 2,500 and 5,000 ppm males and females were less than those of the controls throughout the study. The mean body weights of 10,000 ppm stop-exposure males were generally less than those of the controls throughout the study; those of 10,000 ppm stop-exposure females were less than those of the controls during the exposure portion of the study but increased steadily after the cessation of dosing at week 27. Feed consumption by exposed groups was similar to that by the controls after week 1 of the study. Treatment related eye/nasal discharge, hyperactivity when handled, and/or ataxia were observed in exposed male and female rats on or about day 2 of exposure but were no longer apparent after day 7. Plasma Oxazepam Determinations: Plasma oxazepam concentrations were measured at the end of the study. The concentrations ranged from approximately 0.5 (625 ppm males) to 2.8 &mgr;g/mL (5,000 ppm females). Pathology Findings: In the standard histopathologic evaluation, the incidence of renal tubule adenoma was slightly increased in male rats exposed to 2,500 ppm and was at the upper limit of the historical control range for this neoplasm in 2 year NTP feed studies. In an extended evaluation (step section) of the kidneys of male rats, the incidences of renal tubule adenoma occurred with a positive trend in exposed groups. In standard and step sections (combined), male rats exposed to 2,500 or 5,000 ppm showed a significant increase in the incidences of renal tubule adenoma and hyperplasia. In addition, the incidences of renal tubule adenoma and hyperplasia were significantly increased in the 10,000 ppm stop exposure group. The incidences of nephropathy in continuously exposed female rats were significantly greater than in the controls, and the severity of nephropathy increased wised with increasing exposure concentration in males. The incidences of epithelial hyperplasia and chronic inflammation of the forestomach in males exposed to 2,500 and 5,000 ppm and of ulcers in 2,500 ppm males were significantly greater than in the controls. Incidences of mineralization of the glandular stomach in 5,000 ppm and 10,000 ppm (stop-exposure) males and of erosion of the duodenum in 5,000 ppm males were significantly greater than in the controls. Female rats exposed to 2,500 ppm had greater incidences of epithelial hyperplasia, chronic inflammation, and ulcers of the forestomach and of erosion in the glandular stomach. Centrilobular hepatocyte hypertrophy occurred more frequently in 2,500 and 5,000 ppm males and females than in the controls. GENETIC TOXICOLOGY: Oxazepam was not mutagenic in any of several strains of S. typhimurium, nor did it induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells. These in vitro tests were performed with and without S9 metabolic activation. Results from an in vivo mouse peripheral blood micronucleus test performed on B6C3F1 mice used in a 14-week study were also negative. CONCLUSIONS: In summary, under the conditions of these 2-year dosed-feed studies, there was equivocal evidence of carcinogenic activity in male F344/N rats, based on small increases in the incidences of renal tubule adenomas in exposed groups also exhibiting significantly enhanced nephropathy. There was no evidence of carcinogenic activity of oxazepam in female F344/N rats exposed to feed containing 625, 2,500, or 5,000 ppm for 2 years or 10,000 ppm for 6 months. Administration of oxazepam to rats resulted in nonneoplastic lesions in the forestomach, glandular stomach, and small intestine as well as centrilobular hypertrophy of hepatocytes in the liver. In addition, nephropathy was increased in incidence in female rats and was markedly increased in severity in male rats, resulting in early mortality at the higher exposure concentrations. Synonym: 7 Chloro-1,3-dihydro-3-hydroxy-5-phenyl-2H-1,4-benzodiazepin-2-one Trade Names: Tazepam, Wy-3498, Serax PMID- 12579206 TI - NTP Toxicology and Carcinogenesis Studies of Chloroprene (CAS No. 126-99-8) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Chloroprene is used almost exclusively in the manufacture of neoprene (polychloroprene). Chloroprene was chosen for study because it is a high-volume production chemical with limited information on its carcinogenic potential and because it is the 2-chloro analogue of 1,3-butadiene, a potent, multi-species, multi-organ carcinogen. Male and female F344/N rats and B6C3F1 mice were exposed to chloroprene (greater than 96% pure) by inhalation for 16 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, Drosophila melanogaster, and B6C3F1 mice (bone marrow cells and peripheral blood erythrocytes). 16-Day Study in Rats: Groups of 10 male and 10 female F344/N rats were exposed to 0, 32, 80, 200, or 500 ppm chloroprene by inhalation, 6 hours per day, 5 days per week, for 16 days. Three 500 ppm males died on day 2 or 3 of the study. Mean body weight gains of 200 ppm males and females and 500 ppm females were significantly less than those of the chamber control groups. On the first day of exposure, rats exposed to 500 ppm were hypoactive and unsteady and had rapid shallow breathing. These effects were also observed to some degree in animals exposed to 200 ppm. After the second day of exposure, the effects in these groups worsened, and hemorrhage from the nose was observed. A normocytic, normochromic, responsive anemia; thrombocytopenia; and increases in serum activities of alanine aminotransferase, glutamate dehydrogenase, and sorbitol dehydrogenase occurred on day 4 in 200 ppm females and 500 ppm males. Kidney weights of 80 and 500 ppm females were significantly greater than those of the chamber control group, as were the liver weights of 200 and 500 ppm females. The incidences of minimal to mild olfactory epithelial degeneration of the nose in all exposed groups of males and females were significantly greater than those in the chamber control groups. The incidence of squamous metaplasia of the respiratory epithelium was significantly increased in 500 ppm males. The incidences of centrilobular to random hepatocellular necrosis in 500 ppm males and 200 ppm females were significantly greater than those in the chamber control groups. 16-Day Study in Mice: Groups of 10 male and 10 female B6C3F1 mice were exposed to 0, 12, 32, 80, or 200 ppm chloroprene by inhalation, 6 hours per day, 5 days per week, for 16 days. All males and females exposed to 200 ppm died on day 2 or day 3 of the study. Mean body weight gains of males exposed to 32 or 80 ppm were significantly less than that of the chamber control group. Mice exposed to 200 ppm exhibited narcosis during exposure and were hypoactive with reduced body tone after the first day of exposure. In general, hematology and clinical chemistry parameters measured for exposed males and females were similar to those of the chamber control groups. Thymus weights of 80 ppm males and females were significantly less than those of the chamber control groups. Liver weights of 80 ppm females were significantly greater than those of the chamber control groups. Increased incidences of multifocal random hepatocellular necrosis occurred in males and females exposed to 200 ppm. Hypertrophy of the myocardium, foci of hemorrhage, and mucosal erosion were observed in three males and three females exposed to 200 ppm. Squamous epithelial hyperplasia of the forestomach was observed in two males and two females exposed to 80 ppm. Thymic necrosis, characterized by karyorrhexis of thymic lymphocytes, was observed in all males and females in the 200 ppm groups. 13-Week Study in Rats: Groups of 10 male and 10 female F344/N rats were exposed to chloroprene at concentrations of 0, 5, 12, 32, 80, or 200 ppm by inhalation, 6 hours per day, 5 days per week, for 13 weeks. One male exposed to 200 ppm died during the study. The final mean body weights and body weight gains of all exposed groups of males and females were similar to those of the chamber control groups. Clinical findings in 200 ppm males included red or clear discharge around the nose and eye region. At week 13, a norm a normocytic, normochromic, and non-responsive anemia occurred in 200 ppm males and females. A thrombocytopenia occurred in 200 ppm males and females on day 2 and in 80 and 200 ppm females on day 22. However, at week 13, platelet counts rebounded and were minimally increased in 200 ppm males and females. On day 2, a minimal to mild increase in activated partial thromboplastin time and prothrombin time occurred in 200 ppm males and females. The 200 ppm males and females also had increased activities of serum alanine aminotransferase, glutamate dehydrogenase, and sorbitol dehydrogenase on day 22; these increases were transient, and by week 13 the serum activities of these enzymes were similar to those of the chamber controls. An alkaline phosphatase enzymeuria occurred in 200 ppm females on day 22; at week 13, an alkaline phosphatase enzymeuria oc-curred in 32, 80, and 200 ppm males and 200 ppm females. At week 13, a proteinuria occurred in 200 ppm males. Liver nonprotein sulfhydryl concentrations in male rats immediately following 1 day or 12 weeks of exposure to 200 ppm and in females exposed to 200 ppm for 12 weeks were significantly less than those of the chamber control groups. Kidney weights of 200 ppm males and females and 80 ppm females were significantly greater than those of the chamber control groups. Sperm motility of 200 ppm males was significantly less than that of the controls. In neurobehavioral assessments, horizontal activity was increased in male rats exposed to 32 ppm or greater and total activity was increased in 32 and 200 ppm males. Increased incidences of minimal to mild olfactory epithelial degeneration and respiratory metaplasia occurred in males and females exposed to 80 or 200 ppm. The incidence of olfactory epithelial degeneration in 32 ppm females was also significantly greater than that in the chamber control group. The incidence of hepatocellular necrosis in 200 ppm females was significantly greater than that in the chamber control group. Scattered chronic inflammation also occurred in the liver of male and female rats in the 200 ppm groups; the incidence in 200 ppm females was significantly greater than that in the chamber control group. The incidences of hemosiderin pigmentation were significantly increased in males and females exposed to 200 ppm. 13-Week Study in Mice: Groups of 10 male and 10 female B6C3F1 mice were exposed to chloroprene at concentrations of 0, 5, 12, 32, or 80 ppm by inhalation, 6 hours per day, 5 days per week, for 13 weeks. All male and female mice survived to the end of the study. The final mean body weight and body weight gain of males exposed to 80 ppm were significantly less than those of the chamber control group. Hematocrit concentrations of females exposed to 32 or 80 ppm and erythrocyte counts of 80 ppm females were significantly less than those of the chamber control group. Platelet counts of 32 and 80 ppm females were also greater than that of the chamber control group. Increased incidences of squamous epithelial hyperplasia of the forestomach occurred in males and females exposed to 80 ppm. 2-Year Study in Rats: Groups of 50 male and 50 female F344/N rats were exposed to chloroprene at concentrations of 0, 12.8, 32, or 80 ppm by inhalation, 6 hours per day, 5 days per week, for 2 years. Survival, Body Weights, and Clinical Findings: Survival of males exposed to 32 or 80 ppm was significantly less than that of the chamber control group. Mean body weights of males exposed to 80 ppm were less than those of the chamber controls after week 93. Masses of the torso were observed during the study in exposed female groups, and these clinical findings correlated with mammary gland fibroadenomas observed at necropsy. Pathology Findings: The incidences of squamous cell papilloma and squamous cell papilloma or squamous cell carcinoma (combined) of the oral cavity in male rats exposed to 32 ppm and male and female rats exposed to 80 ppm were significantly greater than those in the chamber controls and exceeded the historical control ranges. The incidences of thyroid gland follicular cell adenoma or carcinoma (combined) in males exposed to 32 or 80 ppm were significantly greater than that in the chamber control group and exceeded the historical control range. Although the incidences of follicular cell adenoma and follicular cell adenoma or carcinoma (combined) in 80 ppm females were not significantly greater than those in the chamber controls, they did exceed the historical control range for these neoplasms. The incidences of alveolar epithelial hyperplasia of the lung were significantly greater in all exposed groups of males and females than in the chamber control groups. The incidences of alveolar/bronchiolar carcinoma and alveolar/bronchiolar adenoma or carcinoma (combined) in 80 ppm males were slightly greater than those in the chamber control group. Although these neoplasm incidences were not significant, they exceeded the historical control range. The incidence of alveolar/bronchiolar adenoma, although not significant, was greater in 80 ppm females than in the chamber control group. The incidences of multiple fibroadenoma of the mammary gland in all exposed groups of females were greater than that in the chamber control group. The incidences of fibroadenoma (including multiple fibroadenoma) in 32 and 80 ppm females were significantly greater than that in the chamber controls. The incidences of fibroadenoma in the chamber control group and in all exposed groups of females exceeded the historical control range. The severity of nephropathy in exposed groups of male and female rats was slightly greater than in the chamber controls. Renal tubule adenoma and hyperplasia were observed in males and females. Additional kidney sections from male and female control and exposed rats were examined to provide a clearer indication of the potential effects of chloroprene on the kidney. The combined single- and step-section incidences of renal tubule hyperplasia in 32 and 80 ppm males and 80 ppm females and the incidences of adenoma and adenoma or carcinoma (combined) in all exposed males were significantly greater than those in the chamber controls. A slight increase in the incidence of transitional epithelium carcinoma of the urinary bladder was observed in 80 ppm females. In addition, one 32 ppm male had a transitional epithelium carcinoma and one 80 ppm male had a transitional cell papilloma. These findings are noteworthy because no urinary bladder neoplasms have been observed in chamber control male or female F344/N rats. In the nose, the incidences of atrophy, basal cell hyperplasia, metaplasia, and necrosis of the olfactory epithelium in 32 and 80 ppm males and females and of atrophy and necrosis in 12.8 ppm males were significantly greater than those in the chamber control groups. The incidences of chronic inflammation were significantly increased in males exposed to 12.8 or 32 ppm and in males and females exposed to 80 ppm. The incidences of fibrosis and adenomatous hyperplasia in 80 ppm males and females were significantly greater than those in the chamber controls. Generally, lesions in the nasal cavity were mild to moderate in severity. 2-Year Study in Mice: Groups of 50 male and 50 female B6C3F1 mice were exposed to chloroprene at concentrations of 0, 12.8, 32, or 80 ppm by inhalation, 6 hours per day, 5 days per week, for 2 years. Survival, Body Weights, and Clinical Findings: Survival of males exposed to 32 or 80 ppm and of all exposed female groups was significantly less than that of the chamber controls. The mean body weights of 80 ppm females were significantly less than those of the chamber control group after week 75. Clinical findings included masses of the head, which correlated with harderian gland adenoma and/or carcinoma in 32 ppm males and 80 ppm males and fe-males. Dorsal and lateral torso masses of female mice correlated with mammary gland neoplasms in 32 and 80 ppm females and subcutaneous sarcomas in 12.8, 32, and 80 ppm females. Pathology Findings: The incidences of alveolar/bronchiolar neoplasms in the lungs of all groups of exposed males and females were significantly greater than those in the chamber control groups and generally exceeded the historical control ranges. The incidences of multiple alveolar/bronchiolar adenoma and alveolar/bronchiolar carcinoma were increased in all exposed groups of males and females. The incidences of bronchiolar hyperplasia in all exposed groups of males and females were significantly greater than those in the chamber control groups. Male mice had a pattern of nonneoplastic liver lesions along with silver-staining helical organisms within the liver consistent with infection with Helicobacter hepaticus. An organism compatible with H. hepaticus was confirmed with a polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP)-based assay. In NTP studies with H. hepaticus-associated hepatitis, increased incidences of hemangiosarcoma have been seen in the livers of male mice. Therefore, hemangiosarcomas of the liver were excluded from the analyses of circulatory (endothelial) neoplasms in males in this study. Even with this exclusion, the combined occurrence of hemangioma or hemangiosarcoma at other sites was significantly increased at all chloroprene exposure concentrations in males and in 32 ppm females. Incidences of neoplasms at other sites in this study of chloroprene were not considered to have been significantly impacted by the infection with H. hepaticus or its associated hepatitis. The incidences of harderian gland adenoma and harderian gland adenoma or carcinoma (combined) in males exposed to 32 or 80 ppm and females exposed to 80 ppm were significantly greater than those in the chamber controls. The incidences of harderian gland adenoma or carcinoma (combined) in 32 ppm males and 80 ppm males and females exceeded the historical control ranges. The incidences of mammary gland carcinoma and adenoacanthoma or carcinoma (combined) in 80 ppm females were significantly greater than those in the chamber control group. The incidences of mammary gland carcinoma and of adenoacanthoma in 32 and 80 ppm females exceeded the historical control ranges. Multiple mammary gland carcinomas occurred in exposed females. The incidences of hepatocellular carcinoma in all exposed female groups and hepatocellular adenoma or carcinoma (combined) in 32 and 80 ppm females were significantly greater than those in the chamber controls; in the 80 ppm group, the incidence exceeded the historical control ranges for carcinoma and adenoma or carcinoma (combined). The incidence of eosinophilic foci in 80 ppm females was also significantly greater than that in the chamber controls. The incidences of sarcoma of the skin were significantly greater in all exposed groups of females than in the chamber controls. The incidences of sarcoma of the mesentery were also increased in all exposed groups of females. The incidence of squamous cell papilloma in 80 ppm females was greater than that in the chamber controls; the difference was not significant, but the incidence exceeded the historical control range. Males also showed a positive trend in the incidence of squamous cell papilloma of the forestomach. In males and females exposed to 80 ppm, the incidences of hyperplasia of the forestomach epithelium were significantly greater than those in the chamber controls. Carcinomas of the Zymbal's gland were seen in three 80 ppm females, and two carcinomas metastasized to the lung. Zymbal's gland carcinomas have not been reported in control female mice in the NTP historical database. The incidence of renal tubule adenoma in 80 ppm males was greater than that in the chamber controls. Though this difference was not significant, the incidence of this rare neoplasm exceeded the historical control range. The incidences of renal tubule hyperplasia in males exposed to 32 or 80 ppm were significantly greater than that in the chamber controls. Additional sections of kidney were examined from control and exposed males to verify these findings. The combined single- and step-section incidence of renal tubule adenoma in 80 ppm males and the combined incidences of renal tubule hyperplasia in all groups of exposed male mice were greater than those in the chamber controls. The incidences of olfactory epithelial atrophy, adenomatous hyperplasia, and metaplasia in 80 ppm males and females were significantly greater than those in the chamber controls. The incidences of hematopoietic proliferation of the spleen in 32 and 80 ppm males and in all groups of exposed females were significantly greater than those in the chamber controls. Genetic Toxicology: Chloroprene was not mutagenic in any of the tests performed by the NTP. No induction of mutations was noted in any of four strains of S. typhimurium in the presence or the absence of S9 metabolic activation enzymes, and no induction of sex-linked recessive lethal mutations was observed in germ cells of male D. melanogaster treated with chloroprene via feeding or injection. In male mice exposed to chloroprene by inhalation for 12 days over a 16-day period, no induction of chromosomal aberrations, sister chromatid exchanges, or micronucleated erythrocytes in bone marrow or peripheral blood occurred. Results of a second micronucleus assay in male and female mice after 13 weeks of exposure to chloroprene via inhalation were also negative. Conclusion: Under the conditions of these 2-year inhalation studies, there was clear evidence of carcinogenic activity of chloroprene in male F344/N rats based on increased incidences of neoplasms of the oral cavity; increased incidences of neoplasms of the thyroid gland, lung, and kidney were also attributed to chloroprene exposure. There was clear evidence of carcinogenic activity of chloroprene in female F344/N rats based on increased incidences of neoplasms of the oral cavity; increased incidences of neoplasms of the thyroid gland, mammary gland, and kidney were also attributed to exposure to chloroprene. Low incidences of urinary bladder neoplasms in male and female rats and lung neoplasms in female rats may also have been related to exposure to chloroprene. There was clear evidence of carcinogenic activity of chloroprene in male B6C3F1 mice based on increased incidences of neoplasms of the lung, circulatory system (hemangiomas and hemangiosarcomas), and harderian gland; increased incidences of neoplasms of the forestomach and kidney were also attributed to exposure to chloroprene. There was clear evidence of carcinogenic activity of chloroprene in female B6C3F1 mice based on increased incidences of neoplasms of the lung, circulatory system (hemangiomas and hemangiosarcomas), harderian gland, mammary gland, liver, skin, and mesentery; increased incidences of neoplasms of the forestomach and Zymbal's gland were also attributed to exposure to chloroprene. Exposure of male and female rats to chloroprene was associated with increased incidences of alveolar epithelial hyperplasia in the lung; nephropathy; and several nonneoplastic effects in the nose including olfactory epithelial atrophy, fibrosis, adenomatous hyperplasia, basal cell hyperplasia, chronic inflammation, respiratory metaplasia, and necrosis. Exposure of male and female mice to chloroprene was associated with increased incidences of bronchiolar hyperplasia and histiocytic cell infiltration in the lung; epithelial hyperplasia in the forestomach; renal tubule hyperplasia (males only); several effects in the nose including olfactory epithelial atrophy, respiratory metaplasia, and adenomatous hyperplasia; and hematopoietic cell proliferation in the spleen. Synonyms: Chlorobutadiene, 2-chlorobuta-1,3-diene, 2 chloro-1,3-butadiene, -chloroprene PMID- 12579207 TI - NTP Toxicology and Carcinogenesis Studies of Ethylbenzene (CAS No. 100-41-4) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Ethylbenzene is mainly used in the manufacture of styrene. Ethylbenzene is also a major component of mixed xylenes used as solvents in agricultural and home insecticide sprays, rubber and chemical manufacturing, and household degreasers, paints, adhesives, and rust preventives. Ethylbenzene is also used as an antiknock agent in aviation and motor fuels. Ethylbenzene was nominated for study by the National Institute for Occupational Safety and Health (NIOSH) and the Occupational Safety and Health Administration (OSHA) because of its potential for widespread human exposure and because of its structural similarity to benzene and toluene. Male and female F344/N rats and B6C3F1 mice were exposed to ethylbenzene (greater than 99% pure) by inhalation for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, mouse lymphoma cells, cultured Chinese hamster ovary cells, and mouse peripheral blood erythrocytes. In previously reported 13-week toxicity studies in which F344/N rats and B6C3F1 mice were exposed to ethylbenzene by whole body inhalation exposure, no histopathologic changes were observed (NTP, 1992). 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to 0, 75, 250, or 750 ppm ethylbenzene by inhalation, 6 hours per day, 5 days per week, for 104 weeks. Survival, Body Weights, and Clinical Findings Survival of male rats in the 750 ppm group was significantly less than that of the chamber controls. Mean body weights of 250 and 750 ppm males were generally less than those of the chamber controls beginning at week 20. Mean body weights of exposed groups of females were generally less than those of chamber controls during the second year of the study. Pathology Findings In male rats exposed to 750 ppm, the incidences of renal tubule adenoma and adenoma or carcinoma (combined) were significantly greater than the chamber control incidences. In addition, the incidence of renal tubule hyperplasia in 750 ppm males was significantly greater than that in the chamber controls. The findings from an extended evaluation (step section) of the kidneys showed a significant increase in the incidences of renal tubule adenoma and hyperplasia in 750 ppm males and females; the incidence of renal tubule adenoma or carcinoma (combined) was significantly increased in 750 ppm males. The severities of nephropathy in 750 ppm male and all exposed female rats were significantly increased relative to the chamber controls. The incidence of interstitial cell adenoma in the testis of 750 ppm males was significantly greater than that in the chamber control group and slightly exceeded the historical control range for inhalation studies. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were exposed to 0, 75, 250, or 750 ppm ethylbenzene by inhalation, 6 hours per day, 5 days per week, for 103 weeks. Survival, Body Weights, and Clinical Findings Survival of exposed groups of male and female mice was similar to that of the chamber controls. Mean body weights of female mice exposed to 75 ppm were greater than those of the chamber controls from week 72 until the end of the study. Pathology Findings In 750 ppm males, the incidences of alveolar/ bronchiolar adenoma and alveolar/bronchiolar adenoma or carcinoma (combined) were significantly greater than those in the chamber control group but were within the NTP historical control ranges. The incidence of alveolar epithelial metaplasia in 750 ppm males was significantly greater than that in the chamber controls. In 750 ppm females, the incidences of hepatocellular adenoma and hepatocellular adenoma or carcinoma (combined) were significantly greater than those in the chamber control group but were within the historical control ranges. The incidence of eosinophilic foci in 750 ppm females was significantly increased compared to that in the chamber controls. There was a spectrum of nonneoplastic liver changes related to ethylbenzene exposure in male mice, including syncytial alteration of hepatocytes, hepatocellular hypertrophy, and hepatocyte necrosis. rosis. The incidences of hyperplasia of the pituitary gland pars distalis in 250 and 750 ppm females and the incidences of thyroid gland follicular cell hyperplasia in 750 ppm males and females were significantly increased compared to those in the chamber control groups. GENETIC TOXICOLOGY: Ethylbenzene gave little indication of mutagenicity, in vitro or in vivo. No induction of mutations was noted in Salmonella typhimurium strain TA97, TA98, TA100, or TA1535 with or without S9 metabolic activation, and no increases in sister chromatid exchanges or chromosomal aberrations were observed in cultured Chinese hamster ovary cells treated with ethylbenzene, with or without S9. In the mouse lymphoma assay, a significant mutagenic response was noted in the absence of S9, but only at the highest nonlethal dose tested and with accompanying cytotoxicity; the test was not performed with S9. No increases in the frequency of micronucleated erythrocytes were observed in vivo in peripheral blood samples from male and female mice exposed to ethylbenzene for 13 weeks. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was clear evidence of carcinogenic activity of ethylbenzene in male F344/N rats based on increased incidences of renal tubule neoplasms. The incidences of testicular adenoma were also increased. There was some evidence of carcinogenic activity of ethylbenzene in female F344/N rats based on increased incidences of renal tubule adenomas. There was some evidence of carcinogenic activity of ethylbenzene in male B6C3F1 mice based on increased incidences of alveolar/bronchiolar neoplasms. There was some evidence of carcinogenic activity of ethylbenzene in female B6C3F1 mice based on increased incidences of hepatocellular neoplasms. Exposure of male and female rats to ethylbenzene resulted in increased incidences of renal tubule hyperplasia and increased severities of nephropathy. Exposure of male mice to ethylbenzene resulted in increased incidences of alveolar epithelial metaplasia, syncytial alteration of hepatocytes, hepatocellular hypertrophy, hepatocyte necrosis, and thyroid gland follicular cell hyperplasia. In female mice, ethylbenzene exposure resulted in increased incidences of eosinophilic foci of the liver, pituitary gland pars distalis hyperplasia, and thyroid gland follicular cell hyperplasia. Synonyms: EB; ethylbenzol; phenylethane PMID- 12579209 TI - Microsatellite analysis of chromosome 3p region in sporadic renal cell carcinomas. AB - The etiology and progression of renal carcinomas (RCC) is still poorly understood. RCC have been classified into several pathological entities. The most frequent type, clear cell carcinoma, accounts for about 80% of sporadic RCC and shows several chromosome abnormalities documented both by conventional cytogenetics, loss of eterozygosity (LOH) and replication error (RER) studies. In 10 clear cell type sporadic RCC we evaluated LOH and RER using a set of 10 microsatellite markers covering the chromosome 3p region, which has been suggested for interstitial deletions. Electrophoresis was performed by automated sequencer ABI Prism 377 and data were analyzed with Genescan and Genotyper 2.5 softwares. We revealed allelic loss in 48,7% of informative microsatellites and a single case of RER. We found the highest LOH frequency in 3p25-26 region where maps Von Hippel-Lindau (VHL) oncosuppressor gene. In addition, DNA hypermethylation, an alternative mechanism of VHL gene silencing, was evaluated by methylation-specific PCR. However hypermethylation status was not detected in any of our tumor samples. PMID- 12579208 TI - New molecular methods for classification, diagnosis and therapy prediction of hematological malignancies. AB - Normal functions of the cell are based on the precise regulation of various genes. If this strict regulation and the hierarchy of genes becomes upset due to flaws in this system, the result will be cellular dysfunction which eventually may lead to carcinogenic transformation. Two basic challenges of the classification of cancers are the discovery of new molecular markers characteristic to defined disease groups and the classification of already diagnosed or new cases into existing groups. This precise classification may open the door to tailored treatment or project the expected outcome of the disease. Today there is unlimited access available to the databases containing sequences and localization of the genes within the confines of Human Genome project. It provides significant help for the discovery of chromosome abnormalities and systematic analysis of gene expression patterns. This is important not only to understand normal functions of the cells, but it also contributes to the identification of new genes that are characteristic to given disease groups as markers and that are potential drug targets. Until the second half of the twentieth century the study of the function and regulation of genes was based on step-by-step investigation of individual genes. Regarding the fact, that the genomes of an increasing number of organisms have become known in whole or in part, numerous new techniques have been developed that facilitated the systematic analysis of gene functions. The aim of this study is to summarize the new, molecular based possibilities for classification, diagnosis and prognosis of hematological malignancies, as well as to summarize the main results of these areas. PMID- 12579210 TI - Determination of optimal conditions for analysis of p53 status in leukemic cells using functional analysis of separated alleles in yeast. AB - Tumor suppressor p53 is transcription factor that participates in control of many cellular functions. Somatic mutations of the p53 gene are frequently detected in human cancers. Several methods can be used for identification of p53 mutations, including FASAY - functional analysis of separated alleles in yeast. FASAY distinguishes yeast colonies expressing functional p53 protein from colonies producing a dysfunctional p53 protein simply on the basis of color. The validity of the method depends on a low background level. There are several sources of background as PCR-induced point mutations, low quality of RNA and alternative splicing of intron 9 affecting the p53 carboxy-terminus. In the present work we show that FASAY can be successfully used for analysis of mRNA isolated from blood samples that were collected and stored for 24 hours at 0 degrees C without undesired increase of background. We also measured fidelity of several commonly used DNA polymerases and determined the most suitable kinds of Pfu DNA polymerases for FASAY. Reaction conditions described in this report allow routine analysis of p53 status in leukemic cells using FASAY. PMID- 12579211 TI - The expression of c-erbB-1 and c-erbB-2 in Iranian patients with gastric carcinoma. AB - To assess the significance of epidermal growth factor receptor family members, the overexpression of c-erbB-1 and c-erbB-2 was retrospectively investigated in 146 southern Iranian gastric cancer patients. Indirect immunostaining was used to evaluate the expression of these two receptors in formalin-fixed paraffin embedded tissue samples. c-ErbB-1 expression was observed in 47 (32.2%) and c erbB-2 expression was observed in 24 (16.4%) of tumors. Significant positive correlations were observed between c-erbB-1 expression and tumor size, local invasion, lymph node involvement and tumor stage. There was also a negative correlation between c-erbB-2 expression and tumor stage. These results may suggest the contribution of c-erbB-1 molecule in progression of gastric carcinomas in southern Iranian patients. Moreover, the relatively high percentage of c-erbB-2 positive tumors may provide a useful target for the immunotherapy of these cancers. PMID- 12579212 TI - Considerations in the treatment of the node-negative (N0) neck in glottic carcinomas. AB - Treatment of lymph node negative (N0) glottic carcinoma has raised numerous controversy for decades. Prevention is one of the oldest axioms in medicine. On the other hand, overtreatment can cause unnecessary harm to patients. This retrospective study was performed in 206 patients having glottic cancers with clinically node-negative (N0) necks. The aim of this assessment is to deal with the diagnosis, predictive factors and surgical therapy of occult metastases of squamous cell cancers originating from the glottic region. The examinations were performed in three phases. Preoperative clinical, histological - and in selective cases - imaging were carried out to separate high-risk patients. Intraoperative cases of open surgery after U-shaped skin preparation up to the hyoid bone with direct inspection of jugular lymph node chain (JLNCh) where the neck was staged. The enlarged suspicious nodes were submitted for immediate frozen section. The types of neck dissection were based on the size, shape, number and histological diagnosis of regional nodes. The postoperative additional management was decided according to the results of definitive pathological findings from serial sections of the dissected specimen. Endolaryngeal LASER surgery was carried out in 87 patients based on clinical, histological and imaging criteria. In the course of two years follow-up 2 occult metastases became clinically apparent. At 119 cases open surgery were performed. In 51 patients we could not see enlarged lymph nodes (N< 2 mm) with direct examination, and thus the JLNCh remained intact. In 68 patients elective neck dissections (END) were carried out. In cases of extracapsular spread (ECS) and/or multiple nodal involvements additional radiotherapy was given. PMID- 12579214 TI - Bilateral renal cell carcinoma in a horseshoe kidney. AB - We report a case of bilateral renal cell carcinoma in a horseshoe kidney. To the best of our knowledge this is the second reported case in the international literature. We performed different radiological examinations preoperatively to identify of blood supply, because correct preoperative location of vessels is mandatory. PMID- 12579213 TI - General features of gastric carcinomas and comparison of HSP70 and NK cell immunoreactivity with prognostic factors. AB - During the period of 1996-1998 ninety-four gastrectomy specimens with gastric carcinoma referred to Erciyes University, Medical Faculty, Department of Pathology, were examined histopathologically, histochemically and immunohistochemically. General characteristics of gastric carcinomas and prognostic factors were studied. According the Lauren classification, of the 94 cases of gastric carcinomas, 56 were intestinal type, 21 were diffuse type and 17 were mixed type carcinoma. The association rates of Helicobacter pylori, chronic atrophic gastritis and intestinal metaplasia with gastric carcinomas were high. There was strong immunorectivity with HSP70 in 62,5% of the intestinal type carcinomas. This ratios were lower in diffuse and mixed type carcinomas (p<0.05). The more tumor size and invasion depth increased, the more HSP70 immunoreactivity was obtained (p<0.05). HSP70 immunorectivity was considerably higher in the patients having lymph node metastasis and vascular invasion (p<0.05). It was found that the NK cell number was low in the tumor but higher around the tumor in early gastric carcinomas, compared with advanced carcinomas (p>0.05). In the tumors larger than 10 cm with vascular invasion, NK cell number was lower around the tumor (p>0.05). Defining prognostic factors of gastric carcinomas is of importance to clinicians. It is thought that HSP70 immunoreactivity, besides invasion depth, lymph node metastasis, vascular invasion, tumor size and inflammatory reaction against the tumor, is important in prognosis and associated with advanced stage. PMID- 12579215 TI - Atypical histiocytic infiltration simulating diffuse-type carcinoma in a gastric ulcer due to non-steroidal anti-inflammatory drugs. AB - A 83-year old man treated with naproxen during two years was admitted because of hypovolemia and peritoneal irritation. A panendoscopic study was performed and an ulcer localized at the large curvature of the stomach was disclosed. In the gastrectomy specimen the ulcer showed necrosis, edema, fibrosis, chronic inflammatory infiltrate with lymphocytes and plasma cells. Additionally, atypical cells with irregular and hyperchromatic nuclei or vacuolated cytoplasm were seen in the lamina propia and infiltrating the muscular layers; isolated signet-ring like cells were also seen. Histochemical study with periodic acid-Schiff, mucicarmin, and colloidal stains revealed mucosubstances in these cells. A poorly differentiated carcinoma was initially diagnosed. However, the immunohistochemical study were positive for histiocytic markers (CD-68, S-100 protein) and negative for epithelial markers (cytokeratin; and epithelial membrane antigen). The positivity of mucus stains in the histiocytes could be explained in this case by phagocytosis of mucous substances released from broken hyperplastic glands in the vicinity of the ulcer. To our knowledge, atypical histiocytic infiltration in gastric ulcers has not been previously described; thus, it should be included in the group of gastric carcinoma mimicks. PMID- 12579216 TI - Carcinosarcoma of the stomach. AB - In the gastrointestinal tract, carcinosarcomas are most frequently seen in the esophagus. Carcinosarcoma in the stomach is a rare tumor. We report a carcinosarcoma of the antrum of stomach. The tumor was polypoid and exophytic in appearance and located in the antrum. Immunohistochemical studies showed positivity for cytokeratin, epithelial membrane antigen and cytoplasmic carcinoembryonic antigen in the epithelial component. Positive staining with vimentin, desmin and focal smooth muscle actin and negative staining with chromogranin were observed in spindle cells. Nuclear positive staining was observed with p53 and Ki-67 in both glandular and spindle atypical cells. PMID- 12579217 TI - Ciliated hepatic foregut cyst. AB - Ciliated hepatic foregut cyst is a rare, benign, most often solitary and unilocular, rarely multilocular cyst made up of a ciliated pseudostratified columnar epithelium, a subepitheial connective tissue layer, a smooth muscle layer and an outer fibrous capsule. The lesion is usually found incidentally by ultrasonography, during surgical exploration or autopsy. Recent publications characterizes of its fine needle aspiration biopsy features. The lesion is mostly asymptomatic, however one case caused portal vein compression and another which showed malignant trasformation through squamous metaplasia which warns to examine these lesions cautiously. As the lesion is extremely rare it is difficult to estimate its prevalence and its nature, so every single case presentation could be important. PMID- 12579218 TI - Pulmonary lymphomatoid granulomatosis evolving to large cell lymphoma in the skin. AB - Lymphomatoid granulomatosis is an angiodestructive, angioinvasive lymphoproliferative disorder. It involves most frequently lungs, central nervous system and skin. Recent studies indicate that lymphomatoid granulomatosis is an Epstein-Barr virus associated B cell disorder with a background of reactive T lymphocytes. In a 49 year old woman presenting with fever, malaise and pulmonary masses the diagnosis of lymphomatoid granulomatosis was established histologically by open lung biopsy. Following the initial diagnosis the patient was found to have gastric and skin involvement. The skin lesion was diagnosed as diffuse large B-cell lymphoma. PMID- 12579219 TI - Clear cell odontogenic carcinoma: a diagnostic dilemma. AB - A 26-year-old man presented with a swelling in the right side of face and CT scan revealed a destructive tumor in the right maxilla. Tumor recurred within 5 years of its excision and histopathological examination revealed a clear cell odontogenic carcinoma. The rarity of this tumor, occurrence in maxilla and young age of the patient are some of the rare features which need documentation. The importance of its diagnosis and various differential diagnoses are discussed. PMID- 12579256 TI - Role of telomerase in radiocurability (review). AB - Telomerase is a ribonucleoprotein complex that elongates telomeres by adding hexameric (TTAGGG) repeats to the telomeric ends of the chromosomes, thus compensating for the continued erosion of telomeres. Telomerase activity is present in unicellular organisms and germ cells, both places where it is expected to play a role in indefinite cycling and protection from shortening of the telomeres. One phenotypic manifestation that is virtually pathognomonic of several cancer cells is the telomerase activity. Telomerase activity is enhanced in several cell types after treatment with ionizing radiation (IR). Whether there is a direct correlation between the levels of telomerase activity and IR response for tumor cell kill is yet to be addressed in detail. In this review, information is summarized on telomerase activity as a measure for monitoring the radiocurability of tumors. As tumor growth is partly due to deregulated cell cycling, insights into telomerase activity through the cell cycle may prove helpful in designing therapeutic agents either for telomerase activation in cells where its expression can overcome senescence and prolong the life of post-mitotic cells, or inhibition of telomerase where it is essential for proliferation and thus can provide a therapeutic advantage to kill the tumor cells. PMID- 12579257 TI - Reduction of dihydrodiol dehydrogenase expression in resected hepatocellular carcinoma. AB - Dihydrodiol dehydrogenase (DDH) is one of the major enzymes catabolizing polycyclic aromatic hydrocarbons in the liver. Although four DDH isoforms have been detected in the normal liver, only DDH1 and DDH2 have been detected in cancer cells of lung and esophagus. Moreover, the available information about hepatic pathophysiological regulation of DDH expression is limited. Therefore we addressed the question of DDH expression in patients with liver disorders, in particular, patients with hepatocellular carcinoma (HCC). Expression of DDH1/2 was determined by immunohistochemistry, immunoblotting and reverse transcription polymerase chain reaction (RT-PCR) in 52 patients with resected HCC. DDH1/2 expression was detected in 31 (59.6%) of 52 pathological sections. Frequency of DDH1/2 expression was significantly higher in patients with tumor size >2 cm, and in those who had early local recurrence. In addition to the tumor size and frequency of local recurrence, our results further indicated that expression of DDH1/2 was correlated with those of cyclooxygenase 2 (COX-2), interleukin-6 (IL 6), microsomal epoxide hydrolase (mEpH) and soluble epoxide hydrolase (sEpH) in HCC patients. Interestingly, the expression of DDH1/2 was found inversely correlated with that of glutathione S-transferase (GST) and NADPH p450 reductase (NPR). In conclusion, these results indicate that DDH expression was significantly decreased in about 40% of HCC patients. However, in the bordering non-neoplastic region of liver DDH1/2 expression increased, and the increased DDH1/2 expression correlated with tumor size and the disease progression. PMID- 12579258 TI - Comparative analysis of K-ras point mutation, telomerase activity, and p53 overexpression in pancreatic tumours. AB - K-ras point mutation, p53 over-expression, and telomerase activity have been proposed as molecular markers for clinical diagnosis of pancreatic carcinoma. To evaluate the clinical usefulness of these markers, we performed comparative analysis in 61 resected pancreatic samples including 15 intraductal papillary mucinous tumours (IPMTs), 4 mucinous cystic tumours, 37 ductal adenocarcinomas, and five chronic pancreatitis samples. K-ras point mutation, telomerase activity, and p53 overexpression were analyzed using mutant allele specific amplification, the telomeric repeat amplification protocol, and immunohistochemical staining, respectively. In malignant tumours, K-ras mutation, telomerase activity, and p53 overexpression were detectable in 76, 91, and 46%, respectively, while in benign tumours, these alterations were detectable in 38, 0, and 0%, respectively. Among 15 IPMTs, K-ras mutation was detectable in 4 (80%) of 5 IPMT-adenomas, 4 (80%) of 5 IPMT-carcinomas and 2 (66%) of 3 papillary-mucinous carcinomas, which are invasive carcinomas derived from IPMTs. Telomerase activity was not detectable in IPMT-adenomas, but was detected in all 5 IPMT-carcinomas and 3 papillary-mucinous carcinomas. p53 overexpression was not detected in IPMTs, but was detected in 2 (66%) of 3 papillary-mucinous carcinomas, indicating that telomerase is likely to be activated concomitant with carcinogenesis. These results suggest that telomerase activity is the most useful as a differential diagnostic marker between malignant and benign pancreatic tumours. PMID- 12579259 TI - Expression of bcl-2 in oral squamous cell carcinoma: an immunohistochemical study of 90 cases with clinico-pathological correlations. AB - Apoptosis is a genetically determined process playing an active role in tissue size regulation, morphogenesis and removing damaged cells that could be potentially dangerous for their host. Several agents involved in apoptosis regulation, such as the bcl-2 family components, act as oncogenes and are involved in oral carcinogenesis. Aim of this study is to explore bcl-2 immunoreactivity in oral cancers and to assess its potential clinico-pathological implications. Ninety oral squamous cell carcinoma and 10 normal mucosal formalin fixed, paraffin-embedded samples were analysed for bcl-2 expression by immunohistochemistry. Normal oral mucosa showed a cytoplasmic pattern of bcl-2 immunoreactivity in the basal cell layers. Seventy-four cases of carcinoma (83%) showed no immunoreactivity, at variance with 16 cases (17%) manifesting consistent cytoplasmic positivity. Overall, the peripheral cells of differentiating epithelial tumour islands were intensely stained, with decreasing immunoreactivity toward the centre of the neoplastic nests. Fully keratinised tumour cells showed inconspicuous or absent bcl-2 immunoreactivity. No statistically significant correlations could be demonstrated between bcl-2 immunoreactivity and the sex of the patients, tumour size and with the occurrence of lymph node metastases. Though a direct correlation was found between bcl-2 immunoreactivity and increasing tumour stage, this did not reach statistical significance. Furthermore, G1 and G3 tumours displayed higher percentages of bcl 2-positive cells in comparison with G2 neoplasms and the different distribution of bcl-2 immunoreactivity in G2 and G3 was statistically significant (p<0.05). Finally, patients with absent or low (scores 0 and 1) bcl-2 immunoreactive tumours manifested poorer overall survival rates in comparison with patients with moderate or high (scores 2 and 3) bcl-2 immunoreactive tumours but the difference was not statistically significant. In normal oral mucosa bcl-2 protein is selectively present in the basal cell layers and possibly participates in the control of the terminal keratinocytes differentiation. The study of bcl-2 immunoreactivity possibly may be useful for better characterising and predicting the prognosis of oral SCC but cooperative studies are needed to assess its applications in the clinical practice. PMID- 12579260 TI - VBP-1 is necessary for morphogenesis in Caenorhabditis elegans. AB - VBP-1 has been isolated as a novel protein binding to the von Hippel-Lindau (VHL) tumor suppressor gene product. The fundamental function of the gene was examined in the nematode, Caenorhabditis elegans (C. elegans). The C. elegans VBP-1 gene was expressed in adults only in the gonads and was also expressed in early stage embryos. Double-stranded RNA interference (RNAi) against VBP-1 resulted in an arrest of embryogenesis at morula stages. This suggests that the VBP-1 product is necessary for morphogenesis. PMID- 12579261 TI - Cyanidin 3-O-beta-D-glucoside isolated from skin of black Glycine max and other anthocyanins isolated from skin of red grape induce apoptosis in human lymphoid leukemia Molt 4B cells. AB - Cyanidin 3-O-beta-D-glucoside (CG) was purified from black bean seed coat and other anthocyanins were prepared from red grape skin. These anthocyanins were identified by Mass, and 1H- and 13C-NMR. The effects of four anthocyanins on cell viability in human leukemia Molt 4B cells were investigated. The anthocyanins displayed strong growth inhibitory effects against human leukemia Molt 4B cells. Morphological changes showing apoptotic bodies were observed in the Molt 4B cells treated with these anthocyanins. The fragmentations by these anthocyanins of DNA to oligonucleosomal-sized fragments, that is a characteristic of apoptosis, were observed to be concentration-dependent. N-acetyl-L-cysteine, an antioxidant, suppressed the fragmentation of DNA by these anthocyanins. These findings suggest that growth inhibition of Molt 4B cells by these anthocyanins result from the induction of apoptosis by these compounds and that active oxygen is involved in the induction of apoptosis in the Molt 4B cells. PMID- 12579262 TI - Papillary renal cell carcinoma in a young adult: serial changes in diagnostic images. AB - In renal tumors, we often encounter cases in which accurate diagnosis before surgery is difficult. The serial changes in various diagnostic images were studied in a case of renal cell carcinoma in which definitive diagnosis could not be made by various imaging examinations and biopsies. We report on a case of renal cell carcinoma in a young adult. A 19-year old man complained of left flank pain and pyrexia. Computerized tomography (CT) scan and urography revealed a solid mass in the left kidney. Angiography revealed no abnormality, and CT scan after 3 months showed that the mass was decreasing in size, which suggested an inflammatory mass. CT scan and retorograde pyelography 12 months later showed that the tumor had enlarged and that much of the tumor had extended into the renal pelvis. Left nephrectomy was performed and histological examination revealed that the tumor was papillary renal cell carcinoma. When imaging results and clinical symptoms are atypical, pre-surgery diagnosis of renal cell carcinoma is extremely difficult. In such cases, we found that it was effective to observe the changes in various diagnostic images and to make an overall judgment based on these results. PMID- 12579263 TI - Expression of a novel splicing variant deleting exons 4 and 6 of the progesterone receptor gene is a rare event in breast cancer. AB - We identified a novel alternatively spliced isoform of PR mRNA in breast cancer tissues. The deleted transcript was characterized by an out-of-frame deletion of 437 bp, corresponding to the complete loss of exons 4 and 6 (PR delta4+6 ASV). PR delta4+6 ASV will result in a partial defect in the region of the ligand-binding domain of hormone receptors, suggesting that the conserved residues are missing from the core of the protein. In the limited number of samples studied, a novel PR delta4+6 mRNA was detected in 1 of 45 (2.2%) non-cancerous tissues of patients with breast cancer, in 5 of 45 (11.1%) cancerous tissues of patients with breast cancer. Loss of both exons 4 and 6 will be induced by incomplete splicing and/or repair mechanism. Further studies are necessary to establish the biological significance of this alternative splicing. The expressions of ASVs that induced the mimic PR transcripts need to be considered when designing strategies for regulation analysis of the PR gene. PMID- 12579265 TI - Sentinel lymph node navigation surgery for pancreatic head cancers. AB - Recently, sentinel lymph node (SN) concept has been validated for gastrointestinal and breast cancers. Our previous study has shown that the No. 13 posterior pancreaticoduodenal lymph node group constitutes the major regional drainage site from primary tumors in the pancreatic head, and that the status of these nodes predicts that of the No. 16 abdominal paraaortic lymph node group. Based on these results, we have developed SN navigation surgery for pancreatic cancer, in the search for more curable and less invasive surgery. In brief, 2% patent blue dye is injected into the peritumoral area. Approximately 5 min later, one or more blue-stained nodes within the area of the No. 13 lymph node group are identified and excised for intraoperative frozen section examination. The subsequent surgical decision-tree is as follows: i) if No. 13 SNs are negative, an extended No. 16 lymph node dissection is not performed to reduce morbidity, and ii) when cancer is found, the No. 16 lymph nodes are dissected completely. Since July 1997, nine of 21 patients scheduled to undergo an extended curative surgery underwent SN biopsy. SNs within the area of the No. 13 lymph node group were identified in 8 (89%) patients. An extended No. 16 lymph node dissection was avoided in 4 SN-negative patients. The overall 3-year survival rate of the 21 patients was 36%, and 4 patients (three SN-negative and one SN-positive patients) with stage IVa disease were alive 3 years after surgery. Three SN-negative patients underwent an extended curative pylorus-preserving pancreaticoduodenectomy (PpPD) with combined portal vein resection, but without an extended No. 16 dissection. In conclusion, SN biopsy and curative PpPD can increase curability, reduce morbidity, and provide long-term survival in patients with locally advanced pancreatic head cancer as an alternative to routine extended No. 16 lymph node dissection. PMID- 12579264 TI - Prognostic significance of CD8+ T cell and macrophage peritumoral infiltration in colorectal cancer. AB - The purpose of this study was to evaluate the prognostic significance of CD8+ T cell and macrophage peritumoral infiltration in patients with colorectal cancer. A total of 97 adenocarcinomas of the colon and rectum were examined. Immunohistochemical staining was performed by the standard avidin-biotin peroxidase complex method using antibodies to CD8 and CD68. Peritumoral infiltration by CD8+ T cells or macrophages was evaluated along the invasive margin of the cancer in each specimen. The area with the most abundant infiltration was selected, and the number of immunoreactive positive cells counted at x400 magnification. Patients were divided into two groups based on the degree of infiltration by each cell type: namely those with a high level of infiltration (more than the mean number of positive cells) and those with a low level of infiltration (less than the mean number of positive cells). Patients with a low level of macrophage infiltration had a significantly deeper depth of invasion than patients with a high level of macrophage infiltration (P=0.027). The percentage of patients with a high level of macrophage infiltration was significantly higher in vascular invasion-negative cases (46.7%) than in vascular invasion-positive cases (22.7%; P=0.045), and in lymph node metastasis-negative cases (52.9%) than in lymph node metastasis-positive cases (28.3%; P=0.014). Overall survival was significantly shorter for patients with a low level of CD8+ T cell infiltration than those with a high level of CD8+ T cell infiltration (P=0.01). The survival rate for patients with a high level of both CD8+ T cell and macrophage infiltration was 100%. In conclusion, both CD8+ T cell and macrophage peritumoral infiltration indicates anti-tumoral action in patients with colorectal cancer. PMID- 12579266 TI - S-phase kinase-associated protein 2 expression in laryngeal squamous cell carcinomas and its prognostic implications. AB - The F-box protein S-phase kinase-associated protein 2 (Skp2) positively regulates the G1-S transition by controlling the stability of several G1 regulators, such as the cell cycle inhibitor p27kip1. However, the clinical significance of Skp2 in patients with laryngeal squamous cell carcinoma (LSCC) remains unknown. In this study, a potential distribution of Skp2 in LSCC and its clinical implications was investigated by an immunohistochemical study. Overall, Skp2 overexpression was observed in 36.7% (37 of 102) patients and was significantly associated with lymph node metastasis (p=0.002) and was inversely associated with p27kip1 expression (p=0.026). Survival analysis using the Kaplan-Meier method showed that Skp2 overexpression was significantly associated with shorter disease free and overall survival (p=0.0051 and p=0.0002, respectively). When Skp2 expression and p27kip1 expression were combined, patients with both Skp2 overexpression and reduced expression of p27kip1 revealed poorest disease-free and overall survival as compared to the other cases (p=0.0017 and p<0.0001, respectively). Additionally, in early stage (I, II) cases, Skp2 expression was also revealed to possess a significant prognostic factor in overall survival (p=0.0234), but not in disease-free survival (p=0.2055). By multivariate analysis using the Cox proportional hazards model, tumor grade, tumor size, clinical stage and Skp2 expression were independent prognostic factors both in disease-free and overall survival. These findings indicated that Skp2 expression was closely associated with tumor progression and represented an independent marker for prognosis of LSCC. PMID- 12579267 TI - Immunohistochemical analysis of transforming growth factor beta in gallbladder cancer. AB - TGF-beta is highly expressed in various cancer cells, yet its mechanism suppressing the cell cycle fails and cell proliferation accelerates, resulting in carcinogenesis. However, there are only a very few reports on animal experiments or clinical specimens with regard to the TGF-beta in gallbladder cancer. We performed immunohistochemical analysis of TGF-beta expression with regard to cell proliferation, angiogenesis, and tumor cell infiltration in clinical specimens of gallbladder cancer. TGF-beta immunoreactivity was significantly higher in advanced cancer than in early cancer. With regard to Ki-67 labeling index, there was no significant difference between early cancer and advanced one. There was no statistically significant difference of the density of pre-existing blood vessels (CD34) between TGF-beta-positive group and negative one. The density of angiogenic vessels (CD105) was significantly greater in the TGF-beta-positive group than in the negative one. Tumor-associated macrophage infiltration was significantly higher in the TGF-beta-positive group than in the negative one. No statistically significant differences in cumulative survival rate were noted between patients in the TGF-beta-positive and TGF-beta-negative groups. In conclusion, our study revealed that in patients with gallbladder cancer, expression of TGF-beta increases according to cancer progression and strongly influences angiogenesis and macrophage infiltration, which contributes to tumor proliferation, but acts weakly on cancer cells by itself. PMID- 12579268 TI - Histone H4 acetylation and histone deacetylase 1 expression in esophageal squamous cell carcinoma. AB - The alterations of the chromatin structure by histone acetylases (HATs) and histone deacetylases (HDACs) are implicated in the regulation of gene transcription and also in the process of carcinogenesis. HDAC inhibitors have been shown to be potent inducers of growth arrest, differentiation and/or apoptotic cell death of transformed cells and, as a result, they are currently receiving considerable attention as antitumor agents. In this study, we examined the status of histone H4 acetylation and the level of HDAC1 expression in surgically resected specimens of human esophageal squamous cell carcinoma by immunohistochemistry. We herein demonstrate that histone H4 of esophageal carcinoma cells was significantly hyperacetylated in the early stage of cancer invasion and thereafter changed into a hypoacetylated state according to the degree of cancer progression. The cases in which HDAC1 was less expressed in esophageal carcinoma cells than in the normal mucosa significantly increased as the carcinoma invaded into the deeper layers of the esophageal wall. Furthermore, both the hyperacetylation of histone H4 and the high expression of HDAC1 were shown to topologically colocalize in the same tumor. These results suggested that a dynamic equilibrium between the HAT and HDAC activities is disrupted in esophageal carcinoma, thus implying that a certain interaction may exist between the hyperacetylation of histone H4 and the HDAC1 expression. PMID- 12579269 TI - Immunohistochemical analysis of cyclooxygenase-2 and vascular endothelial growth factor in pancreaticobiliary maljunction. AB - Recent studies have elucidated that cyclooxygenase (COX)-2 is strongly related to cancer progression or development by means of its anti-apoptotic effect, enhancement of angiogenesis or decrease of cell-to-cell adhesive activity. However, there is no report on the relationship between COX-2 expression and angiogenesis in pancreaticobiliary maljunction (PBM). We examined the correlation between the overexpression of COX-2 and vascular endothelial growth factor (VEGF) in 65 lesions from 30 patients with PBM immunohistochemically. The positive expression of COX-2 was found in 20% of regenerative epithelium, 11.1% of hyperplasia without atypia, 86.4% of hyperplasia with mild atypia, 75% of dysplasia, and 75% of cancerous lesions. VEGF was highly expressed in 80% of regenerative epithelium, 27.8% of hyperplasia without atypia, 86.4% of hyperplasia with mild atypia, 66.7% of dysplasia, and 75% of cancerous lesions. The positive rate of both COX-2 and VEFG expression was significantly higher in hyperplasia with atypia, dysplasia and cancerous lesions than that in hyperplasia without atypia. In addition, there was a statistically significant correlation between COX-2 and VEGF overexpression among all lesions. In 6 of 8 patients of various histological types, both COX-2 and VEGF were stained in almost exactly the same locations. In addition, there were no significant differences between the degree of inflammatory cell infiltration in the surrounding stroma and the expression of COX-2 and VEGF, respectively. These results demonstrated a strong relationship between COX-2 and VEGF overexpression in PBM. Therefore, chemoprevention via the suppression of angiogenesis by means of COX-2 inhibitor may be effective in PBM. PMID- 12579270 TI - Expression of matrix metalloproteinases in human transitional cell carcinoma of the urinary bladder. AB - Matrix metalloproteinases (MMPs) are associated with invasion and metastasis of several human malignant tumors. Especially so MMP-7, as it is mainly produced by the actual cancer cells. However, the expression of MMP-7 in transitional cell carcinoma (TCC) of the urinary bladder has not been previously investigated. We examined expressions of MMP-7 and MMP-2 in TCC, and compared them with clinicopathological characteristics in TCCs. Tumor samples from 20 patients with TCC of the urinary bladder who had surgery performed at the Osaka City University Medical School Hospital were immunohistochemically stained. Expression of MMP-2 was significantly stronger in advanced stage and high grade tumors than in low stage and low grade tumors. MMP-7 was also expressed in TCC, having a tendency to be more strongly expressed in high than in low grade tumors. However, there was no significant difference of MMP-7 expression between advanced and low stages. This increased expression of MMP-7 in high grade TCC of the urinary bladder may be associated with tumor invasion and metastasis. PMID- 12579271 TI - Identification of markers of possible prognostic value in 57 giant cell tumors of bone. AB - Giant cell tumor of bone (GCT) is a neoplasm characterized by the presence of large numbers of multinucleated osteoclast-like giant cells, together with mononuclear spindle-shaped cells. Although GCT can be considered a benign lesion, it may exhibit a high biological aggressiveness, which is often associated with enhanced osteolytic properties and development of lung metastasis. By selecting different groups of GCT patients, including patients without evidence of relapse after a median follow-up of 114 months and patients who recurred with lung metastasis, this study focused on the analysis of the expression at clinical onset and in the metastasis of a series of markers involved either in bone resorption modulation or in the metastatic process. By using immunohistochemistry, we analyzed the expression of interleukin-6 (IL-6), a cytokine stimulating bone resorption that has been demonstrated to be released by GCT cells. The expression of factors of the urokinase-type plasminogen activation system, including the urokinase-type plasminogen activator (u-PA) and the plasminogen activator inhibitor type 1 (PAI-1), which have been described to be frequently implicated in the process of degradation of the extracellular matrix during the metastatic process, were also analyzed. Finally, since the action of plasminogen activators is facilitated by the presence of specific receptors on cell surfaces, the analysis included also the u-PA receptor (u-PAR). Our results showed that all these proteins were variably either expressed or overexpressed both in primary tumors and lung metastasis. However, both the level of expression and the incidence of overexpression were higher in primary GCT that relapsed and in lung metastasis compared to primary tumors from disease-free patients, suggesting a possible association of these proteins with a higher biologic aggressiveness of GCT cells. The parallel analysis of a group of primary tumors and of their respective lung metastasis demonstrated that the enhanced expression of one or more of these proteins may confer a selective advantage to GCT cells in terms of systemic invasiveness. Therefore, the evaluation of the expression levels of these proteins at the time of diagnosis may be taken into consideration for a classification of GCT into categories characterized by a different risk to relapse. PMID- 12579273 TI - Effect of the number of uninvolved nodes on survival in early breast cancer. AB - A high number of uninvolved axillary nodes was found by some authors to be associated with poor survival in node-negative breast cancer. We searched for confirmation and extend the investigation to node-positive cases, using population data from the SEER Program. Patients selected were women aged 40-69 years, diagnosed 1988-1997, T1-T2 breast cancer, undergoing axillary dissection with 4-35 nodes examined. Survivals were estimated by the product-limit method and were computed on pooled data. Results in node-negative patients (n=37,519) showed a 5-year overall survival from 92% (95% confidence interval: 88-95%) with 4 uninvolved nodes, to 93% (87-98%) with 34 uninvolved nodes. In node-positive patients (n=16,978), the 5-year survival increased from 50% (44-56%) with 0 uninvolved nodes, to 91% (82-100%) with 30 uninvolved nodes. Survival graphs indicated an improvement or a plateau with higher number of uninvolved nodes. The graphs also suggested that the ratio of involved and uninvolved nodes might be correlated with survival. We conclude that there was no evidence of poor outcome associated with a high number of uninvolved nodes. The incidental finding that ratio-based characterization of node involvement might be a prognostic factor will be further investigated. PMID- 12579272 TI - Real-time RT-PCR of tyrosine hydroxylase to detect bone marrow involvement in advanced neuroblastoma. AB - Neuroblastoma (NB) is characterised by the secretion of catecholamines in approximately 95% of patients. Tyrosine hydroxylase is the rate-limiting enzyme in catecholamine biosynthesis pathway. Expression of the tyrosine hydroxylase gene (TH) is regulated in a tissue-specific manner during neonatal development and differentiation, therefore TH mRNA expression is a specific tumour marker for NB. Here we present a real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay using TaqMan technology for detection and quantification of TH mRNA in bone marrow (BM) NB patients. The degree of TH expression was derived from the ratio of the mRNAs of this gene and the reference gene, beta-actin. A ratio greater than 3x10(-2) was considered as positive for TH mRNA presence. Samples were also examined for TH mRNA by first and nested RT-PCR. Seventeen BM samples from 4 patients with disseminated NB (3 stage IV and 1 stage IVs) were evaluated at diagnosis and during treatment. We found a variable degree of TH expression ranging from 0.0344 to 26.3370 in 12/17 positive samples, while no TH mRNA (value lower than 3x10(-2)) was detected in 5/17 samples obtained after consolidation therapy. Our results show a moderate concordance between different qualitative RT-PCR methods and real-time RT-PCR. The real-time RT-PCR results seem to fit better with the natural short-term clinical follow-up of the evaluated patients, with respect to qualitative methods. Real-time TH RT-PCR could therefore be of clinical value for the assessment of a patient's prognosis by monitoring minimal residual disease (MRD). PMID- 12579274 TI - Prognostic value of Bak expression in oral tongue squamous cell carcinomas. AB - Bak is a pro-apoptotic member of the Bcl-2 family of genes that are involved in regulation of programmed cell death. By means of immunohistochemical methods, Bak expression was evaluated in formalin-fixed, paraffin-embedded diagnostic specimens from 83 patients with T1-T4 oral tongue squamous cell carcinomas (SCC). The fractions of tumor cells expressed Bak and their staining intensities were given an expression score. Bak expression was compared with our previous investigated apoptosis-related parameters such as apoptotic index (AI), Bax and Bcl-2 expression, and p53 accumulation. Bak expression correlated positively with Bcl-2 expression (p=0.0001). No significant correlation was found between Bak expression and Bax expression, AI, or p53 accumulation. Patients with Bak expression exceeding the mean value had poorer disease-specific survival when compared with those with values below the mean Bak expression (p=0.01). Cox proportional hazards regression analysis revealed that Bak expression was a significant, independent prognostic variable (p=0.0325). A two-parameter combination of Bak expression and Bax expression, AI, or p53 accumulation revealed an enhanced prognostic potential (p<0.0001) when compared with single parameters. We conclude that Bak expression, particularly in combination with Bax expression, as well as in combination with AI, or p53 accumulation, has prognostic value in tongue SCC. PMID- 12579275 TI - A water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi) mycelia suppresses azoxymethane-induction of colon cancers in male F344 rats. AB - The present study was designed to investigate the protective effect of a dietary water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi or Mannentake) mycelia (designated as MAK) on the induction and development of azoxymethane (AOM)-induced colon tumors in male F344/Du Crj rats. A total of 80 animals were divided into five groups at six weeks of age, groups 2, 3 and 4 being given weekly subcutaneous injections of AOM (15 mg/kg body weight) for the initial 3 weeks to induce colon tumors. Rats in group 1 and 5 were injected with the vehicle, 0.9% (w/v) saline, following the same schedule. Rats in groups 1, 2, 3, 4 and 5 were fed MF, MF, 1.25% MAK, 2.5% MAK and 2.5% MAK diets, respectively, starting 1 week before AOM treatment and throughout the six-month experimental period. There were no significant differences in number of ACF, total AC and AC per site among groups 2 to 4, but the tumor incidence was significantly lower, and tumor size was smaller in group 4 (AOM + 2.5% MAK) than in group 2 (AOM + MF). Additionally, beta-catenin positive tumor cell nuclei were significantly decreased in the MAK-fed rats (groups 3 and 4), which also demonstrated lowering of the PCNA labeling index and a shortened germinal region in the colon. The present results thus indicate that dietary MAK could act as a potent chemopreventive agent for colon carcinogenesis. PMID- 12579276 TI - Preoperative combination chemotherapy with S-1 and low dose cisplatin against highly advanced gastric carcinoma. AB - We performed preoperative chemotherapy with S-1 and low-dose CDDP (TSLD) to treat 9 patients of highly advanced gastric cancer. Five cases showed significant effect with severe fibrosis diagnosed as grade 1b-2 effect in histology, and all 9 cases showed at least grade 1a effect in either primary lesion or lymph node. While toxicity of less than grade 3 (gastrointestinal disorder and/or bone marrow suppression) were observed in 5 out of 9 cases (55.6%), no symptom was observed in 3 out of the 5 patients. We conclude that the TSLD is useful to control highly advanced gastric cancer. PMID- 12579277 TI - A new thin-type bronchoscope improves diagnostic accuracy of peripheral pulmonary carcinoma. AB - We investigated the XBF-4B40 (4B40) thin bronchoscope, which has a 4.0-mm outer diameter and a 2.0-mm biopsy channel, and studied its impact on the diagnosis of lung carcinoma. We analyzed 30 peripheral lung carcinoma cases by performing bronchoscopic examination using the 4B40 and the type 200 videoscope and recorded the most distal bronchus level reached. In addition, transbronchial aspiration biopsy, forceps biopsy, or brushing was performed, and the diagnosis accuracy between the two instruments was determined. The mean of the most distal bronchus reached by the 4B40 and the type 200 videoscope was the 4.5th and the 3rd, respectively. The 4B40 reached significantly further into the distal bronchus than the type 200 videoscope (p<0.0001). Accuracy for lesions detected in the right segment 1 or left segment 1+2 using the 4B40 and the type 200 videoscope was significantly different (p<0.0001) at 90.0 and 0%, respectively. The new type of bronchoscope, XBF-4B40, is useful for peripheral pulmonary lesion diagnosis with diagnosis accuracy improvements in the right segment 1 or the left segment 1+2. PMID- 12579278 TI - The preventive effect of antineoplaston AS2-1 on HCC recurrence. AB - Once hepatocellular carcinoma (HCC) develops, it repeats intrahepatic metastasis and has multicentric occurrence, which requires frequent treatment. We designed a phase II clinical trail to clarify whether antineoplaston AS2-1, a mixture of sodium salts of phenylacetylglutamine and phenylacetic acid at a ratio of 1:4, prolongs the recurrence-free interval of HCC patients who undergo frequent treatments for recurrence. Ten patients were enrolled in this trial, 2 in stage I, 6 in stage II, 1 in stage III, 1 in stage IV-B at initial diagnosis. Ten patients experienced 35 recurrence-free intervals. Recurrence-free intervals during antineoplaston AS2-1 administration were significantly longer than those without antineoplaston AS2-1 (16.19+/-15.916 versus 5.05+/-2.897 months: p<0.01). Patients who experienced recurrence-free intervals with and without antineoplaston AS2-1 showed longer intervals during antineoplaston AS2-1 administration than those before and after antineoplaston AS2-1 administration (14.47+/-13.821 versus 5.07+/-2.989 versus 5.02+/-3.009 months: p<0.05). Two patients in stage I showed longer recurrence-free intervals than those in more advanced stages. In conclusion, antineoplaston AS2-1 could not prevent recurrence of HCC but prolonged the recurrence-free interval between regional treatments and improved survival rate of these patients. PMID- 12579280 TI - Expression of tetraspanin adaptor proteins below defined threshold values is associated with in vitro invasiveness of mammary carcinoma cells. AB - Tetraspanins are transmembrane adaptor proteins involved in the regulation of various fundamental cellular processes. For a number of malignant diseases, the level of expression of members of the tetraspanin family was found to correlate with tumor cell invasiveness, ability to form metastases, and poor clinical outcome. We describe the exact quantification of mRNAs coding for the tetraspanins CD9, CD63, CD82 and CD151 expressed by mammary carcinoma-derived cell lines that were classified as invasive or non-invasive according to their ability to penetrate collagen-fibroblast gels in vitro. The mean of beta2 microglobulin-normalized expression of CD9 was about 10-fold higher than the mean calculated for CD63 and about 20-fold higher than expression of CD82 and CD151. Direct comparison of tetraspanin expression of invasive and non-invasive cell lines with the Mann-Whitney test revealed a significant correlation for CD63. Grouping of cell lines in relation to threshold values of expression resulted in significant correlations for CD63 (Fisher's exact test p=0.004) and CD151 (p=0.02) but not for CD82 (p=0.065) and CD9 (p=0.168). Expression of CD9, C63 and CD151 was found to be coupled whereas CD82 was expressed independently. This highly significant association points to common mechanisms of gene regulation for this subgroup of tetraspanins. We showed that on basis of absolute amounts of tetraspanin mRNAs, at least in vitro invasiveness is clearly predictable. Our results support the assumption that downregulation of tetraspanins in breast cancer cells is an important step of tumor progression to more malignant phenotypes and underline their important role as mediators in multimolecular membrane protein complexes regulating cell adhesion and migration. PMID- 12579279 TI - External beam radiotherapy in bone metastatic prostate cancer: impact on patients' pain relief and quality of life. AB - Bone metastases are a severe problem in oncology, since they usually are associated with pain. External beam radiation therapy (EBRT) has been, for many years, an important component of the treatment regimen to relieve pain. We have performed a clinical study to evaluate the relationship of response to EBRT in terms of pain relief and improvement in quality of life (QoL). We were also interested in the incidence of acute toxicity with EBRT. We have prospectively evaluated 75 patients (median age 68 years, range 64-79 years) with bone metastases from prostate cancer treated with EBRT, radiographically documented from June 1999 to September 2000. Additional therapies in these patients included: second-line hormonal therapy (HT) in 20 patients, chemotherapy (CT) in 25 patients, biphosphonates in 45 patients. For all patients a pain and narcotic evaluation was done before entering the study. Assessment of response was carried out by evaluating pain relief. QoL was measured by using the EORTC QLQ-C30 questionnaire. Toxicity analysis was based on the ROTG grading system. Survival was calculated by Kaplan-Meier estimate from the start of EBRT treatment until the last follow-up or death. A total of 61 out of 75 patients (81%) experienced some type of pain relief after treatment. The complete response rate was 23%. No significant difference in the response rates was found between the group treated with EBRT alone versus the groups treated with EBRT + CT or EBRT + HT. We noted a significant improvement in some of the scales of the considered EORTC-QLQC30 questionnaire. As expected all treatment-related complications were either grade 1-2 acute or subacute and transitory in nature. The group treated with EBRT + CT experienced slightly higher toxicity rates. There were no treatment-related fatalities. Forteen patients of 61 (23%) responders was alive at 12 months. Our results confirm the ability of EBRT to relieve bony pain in the majority of the cancer patients treated as measured by prospective analysis of pain scales prior to and after EBRT. Minimal side effects were experienced and QoL improved as shown by the results of the specific questionnaire. PMID- 12579281 TI - Increased serum midkine concentration as a possible tumor marker in patients with superficial esophageal cancer. AB - Midkine, a heparin-binding growth factor, is expressed in numerous cancer tissues and is reportedly elevated in patients with various neoplasms. The aim of this study was to evaluate the clinicopathological significance of serum midkine concentration (S-MK) in patients with superficial esophageal squamous cell carcinoma (SCC). Pretreatment S-MK was measured by enzyme-linked immunosorbent assay in 135 healthy controls, 16 patients with benign esophageal disease, and 60 patients with primary superficial esophageal squamous cell cancer (SESCC). All patients with SESCC underwent curative resection. The disease was staged according to TNM/UICC guidelines. Serum concentrations of carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC-Ag), and cytokeratin 19 fragment (CYFRA21-1) were also evaluated in the same populations. S-MK in patients with SESCC (388+/-411 pg/ml) was significantly higher than in benign esophageal disease or healthy controls (183+/-73 and 154+/-76 pg/ml, respectively). Using the mean + 2 standard deviations of healthy control S-MK (300 pg/ml) as the cut-off level, 50% of patients with esophageal SESCC were deemed positive. This S-MK positivity rate for detecting SESCC was significantly higher than for other tumor markers. Thus, S-MK may be useful as a tumor marker to detect SESCC. PMID- 12579282 TI - Thermoradiotherapy combined with adenoviral p53 gene therapy in head and neck squamous cell carcinoma cell lines. AB - We examined effects of recombinant p53-expressing adenovirus combined with thermoradiotherapy in 8 head and neck squamous cell carcinoma (SCC) cell lines to improve the outcomes of the treatment of advanced head and neck cancer. The p53 gene therapy did not improve the discrepancy between thermoradiosensitivities among the 8 SCC cell lines. However, p53 gene therapy improved the effects of thermoradiotherapy in all 8 cell lines, and there were significant differences in four situations of the HSC4 44 degrees C (p=0.032), SAS at 44 degrees C (p=0.029), the KB at 43 degrees C (p=0.025), and the Ca9-22 43 degrees C (p=0.020). In comparing the survival rates of thermoradiotherapy with those of thermotherapy and radiotherapy, thermoradiotherapy demonstrated actual survival rates less than theoretical survival rates based on the survival rates of thermotherapy multiplied by the survival rates of radiotherapy in almost all treatments of thermoradio-gene therapy of the 8 SCC cell lines. These results demonstrate that thermoradiotherapy combined with p53 gene therapy may be a useful tool in treating SCC cells. PMID- 12579283 TI - Promotion effects of hot water on N-nitrosomethylbenzylamine-induced esophageal tumorigenesis in F344 rats. AB - This study is to determine the effects of hot water on N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal tumorigenesis model. F344 rats received one treatment of hot water 1 ml/kg and NMBA 1 mg/kg, or a combination treatment of NMBA 1 mg/kg pus hot water 1 ml/kg, or/and EGCG (epigallocatechin-3-gallate) 10 mg/kg. The experiment was concluded at the 20th week. Our results showed that the number of tumors and incidence of carcinomas were significantly increased by hot water (65 degrees C) (p<0.05, p<0.03, respectively), as compared with the group which received NMBA injections only. EGCG treatment did not significantly reduce the number or the size of tumours as the temperature of added hot water increased. In addition, PGE2 production was induced by NMBA, and further significantly increased by added hot water (p<0.05). On the other hand, EGCG slightly decreased the elevated PGE2 production, however, this effect of EGCG was offset by hot water. Our study further confirmed that the drinking of hot beverages increased the risk of esophageal carcinogenesis, and the drinking hot tea will abolish the inhibitory effects of EGCG on this disease. PMID- 12579284 TI - Immunohistological study of cell cycle-related factors, oncogene expression, and cell proliferation in adenocarcinoma developed in Barrett's esophagus. AB - In addition to presenting clinicopathological findings in 3 patients with adenocarcinoma developed in Barrett's esophagus, we have investigated the expression of cell cycle-related factors, oncogenes and cell proliferation in normal squamous epithelium, specialized columnar epithelium (SCE) and adenocarcinoma in Barrett's esophagus, using immunohistological techniques. The expression of p21 in adenocarcinoma in Barrett's esophagus tended to be decreased in two mutated p53-strongly-positive patients and to be increased in one mutated p53-weakly-positive patient. Furthermore, mutated p53 was strongly expressed in the deep layer of the cancer, while p21 was expressed in the superficial layer of the cancer. Thus, mutated p53 was inversely correlated with p21 in adenocarcinoma in Barrett's esophagus. The mean positive cell rate (PR) of Ki-67 was 4% in normal squamous epithelium, 24.5% in the SCE, and 41.7% in the adenocarcinoma in Barrett's esophagus. The mean PR of proliferating cell nuclear antigen (PCNA) was 6% in normal squamous epithelium, 29.5% in the SCE, and 55% in the adenocarcinoma in Barrett's esophagus. Thus, the PR of Ki-67 and PCNA were clearly higher in the SCE in Barrett's esophagus than in normal squamous epithelium, indicating increased cell proliferation in the SCE in Barrett's esophagus. In conclusion, mutated p53 was inversely correlated with p21 in adenocarcinoma in Barrett's esophagus. p53 mutation and the expression of oncogenes such as c-erbB-2 and MDM2 were observed in the SCE in Barrett's esophagus, which showed higher cell proliferation than normal squamous epithelium, suggesting a high malignant potential of the SCE in Barrett's esophagus. We considered that it was important to carefully follow-up patients with Barrett's esophagus. PMID- 12579285 TI - Neoadjuvant chemotherapy with CDDP and 5-fluorouracil for gastric cancer with serosal invasion. AB - Many gastric cancer patients who recur peritoneally are initially diagnosed with serosal invasion. To clarify the usefulness of neoadjuvant chemotherapy with 5 fluorouracil (5-FU) +/- cisplatin (CDDP), neoadjuvant versus no preoperative chemotherapy for gastric cancer with preoperative serosal invasion was investigated. The patients were treated preoperatively with 5-FU 300 mg/m(2)/day for 2 weeks (F group; n=40), 5-FU 300 mg/m(2)/day for 2 weeks + CDDP 15 mg/m(2)/day for 2 days (FP group; n=80) or nothing (C group; n=100). A total of 78% of patients in C, 65.0% in F and 67.5% in FP group were classified as T3 or higher surgically. In patients without peritoneal metastasis, the positive peritoneal lavage cytology was 29.2% in C, 11.8% in F, and 12.2% in FP patients (p=0.0279). Serosal invasion was found histologically in 60.0% of C, 30.0% of F, and 33.8% of FP patients (p=0.001). There were no serious drug reactions and no increases in morbidity or mortality using either regimen. The 5-year survival rate was 47.0% in F and 50.9% in FP patients, but only 33.2% in C patients (p=0.0042). In conclusion, neoadjuvant chemotherapy with 5-FU +/- CDDP for gastric cancer patients with serosal invasion may reduce positive peritoneal cytology, eliminate cancer cells from the serosal surface, and improve prognosis. PMID- 12579286 TI - Clinicopathologic characteristics related to the high variability of coding mononucleotide repeat sequences in tumors with high-microsatellite instability. AB - Frameshift mutation at coding mononucleotide repeat sequences are common in tumors with microsatellite instability (MSI-H), but the incidences are different among the target genes. We analyzed the mutational profiles of 12 known target genes containing polydeoxyadenosine repeats in their coding sequences in 39 MSI-H colorectal carcinomas and 40 MSI-H gastric carcinomas by using polymerase chain reaction and sequencing, and compared the results with the clinicopathologic characteristics. Frameshift mutations of target genes in the MSI-H colorectal and gastric carcinomas are increased according to the length of the polydeoxyadenosine repeats in the target genes. The mean mutational rates of MSI H colorectal carcinomas and MSI-H gastric carcinomas were 2.03 and 1.95 in the 4 genes containing (A)10 repeats, 1.23 and 0.73 in the 4 genes with (A)9 repeats and 0.61 and 0.48 in the 4 genes containing (A)8 repeats, respectively (p<0.001). Among the evaluated clinicopathologic findings, intestinal type gastric carcinomas had more frameshift mutations than the diffuse type carcinomas (3.5 vs. 1.9, p=0.01). These findings suggest that mutational rates of the target genes in MSI-H tumors are diverse, and higher mutational rates are related to the length of mononucleotide repeat sequences of the target genes and histologic type of tumors. PMID- 12579287 TI - Co-expression of receptor for advanced glycation end products and the ligand amphoterin associates closely with metastasis of colorectal cancer. AB - The clinical relevance of the receptor for advanced glycation end products (RAGE) and amphoterin expression was examined in 119 non-diabetic patients with colorectal carcinoma. Expression of RAGE was examined by immunohistochemistry and that of amphoterin by in situ hybridization. RAGE expression was observed in 55% (64/119) of the cases. RAGE positivity in Dukes' B, C and D cases was 19, 81 and 100%, respectively (p<0.0001). Amphoterin was expressed in most cases regardless of tumor stage. Survival analysis of Dukes' B and C cases showed a significantly poorer prognosis for patients with co-expression of RAGE and amphoterin than for patients without co-expression. The results suggest that co-expression of RAGE and amphoterin is closely associated with invasion and metastasis of colorectal cancer. PMID- 12579288 TI - Epidemiology and pathogenesis of esophageal cancer: management and its controversial results (review). AB - Esophageal cancer including squamous cell and adenocarcinoma is considered to be a serious malignancy with respect to prognosis and a fatal outcome in the great majority of cases. Epidemiological data define a certain geographical distribution and relate pathogenesis and aetiology to several chemical substances as well as to Barrett's esophagus and reflux. Surgery remains the standard treatment for localized disease and offers approximately a 10% 5-year survival. Radiation therapy and chemotherapy have been exploited during the last decades not only in advanced cases but also in addition to surgery in a combined modality, in order to increase the cure rate. The controversial results of trials dealing with the management of esophageal cancer are critically reviewed. PMID- 12579289 TI - Pin1 is overexpressed in oral squamous cell carcinoma and its levels correlate with cyclin D1 overexpression. AB - Phosphorylation of serine or threonine residue preceding proline (Ser/Thr-Pro) is a key regulatory mechanism. The conformation of certain phosphorylated Ser/Thr Pro bonds is regulated specifically by the prolyl isomerase Pin1. Inhibition of Pin1 induces apoptosis and may contribute to neuronal death in Alzheimer's disease. It has been reported that Pin1 is strikingly overexpressed in a subset of human tumors. The differential display screen revealed that Pin1 increases the transcription of several target genes, including cyclin D1 and c-myc genes. Pin1 cooperates with Ras signaling in increasing the transcriptional activity of c-Jun towards cyclin D1. Pin1 also regulates turnover and subcellular localization of beta-catenin by inhibiting its interaction with adenomatous polyposis coli protein (APC). However, the analysis of Pin1 expression has not been demonstrated in human oral squamous cell carcinoma (OSCC). We examined the expression of Pin1 mRNA and protein in OSCC cell lines, and analyzed Pin1/cyclin D1/beta-catenin expression in OSCC clinical samples by immunohistochemical staining. We report that Pin1 is overexpressed in OSCC and its level correlates with cyclin D1 level. These results indicate that Pin1 is related to oncogenesis of OSCC. PMID- 12579290 TI - Determinants of the interval between diagnosis and treatment in patients with digestive tract cancer. AB - The main objective was to analyze the influence of social factors on the interval from diagnosis to treatment in a cohort of 217 patients with digestive tract cancer. From the clinical charts data were obtained for: date of the beginning of illness, dates of medical attendance, type and date of diagnostic tests, characteristics of the disease (symptoms, stage), and date of treatment. From the personal interview we gathered information lacking in the clinical charts such as socio-demographic factors, other underlying diseases, and variables related to life style. The information on hospital admission, diagnosis, and treatment was prospectively gathered, whereas data before hospitalization were retrospectively assessed. In statistical analysis the Kruskal-Wallis test and Cox regression were used. The predictors associated with a shorter diagnosis-treatment period were: low social class, lymphatic involvement at diagnosis, first visit at secondary/tertiary level of health care, no car availability, two or more symptoms at the beginning of disease, sex (male), age (less than 74 years), and diagnosis out of vacation periods. In conclusion, several socio-demographic variables (age, gender, social class, and car availability) influence the interval diagnosis-treatment. PMID- 12579291 TI - Resistance to cytotoxic chemotherapy-induced apoptosis in human prostate cancer cells is associated with intracellular clusterin expression. AB - We recently reported the powerful antiapoptotic activity of clusterin against various apoptotic stimuli in prostate cancer model systems; however, the precise mode of clusterin action in target cells remains largely unknown. In the present study, we therefore investigated whether intracellular or extracellular action of clusterin plays a crucial role in cytotoxic chemotherapy-induced apoptosis in androgen-independent human prostate cancer PC3 cells, which express a high level of clusterin. The sensitivity of PC3 cells to paclitaxel was increased by pretreatment with monoclonal antibody (mAb) to clusterin or antisense (AS) oligodeoxynucleotide (ODN) targeting the clusterin gene in a dose-dependent manner at up to 50 microg/ml or 1 microM, respectively. However, clusterin mAb failed to further enhance the sensitivity to paclitaxel of PC3 cells simultaneously treated with 1 microM AS clusterin ODN, whereas AS clusterin ODN further induced the apoptosis of cells treated with 50 microg/ml clusterin mAb. Moreover, the effects of clusterin mAb and AS clusterin ODN on PC3 cells were not reversed by additional treatment with exogenous recombinant clusterin protein. These findings suggest that the sensitivity of PC3 cells to paclitaxel-induced cytotoxicity may be regulated by the intracellular rather than extracellular level of clusterin. PMID- 12579292 TI - Evaluation of different markers in non-small cell lung cancer: prognostic value of clinical staging, tumour cell detection and tumour marker analysis for tumour progression and overall survival. AB - We compared the prognostic value of routine pathology, cytokeratin-positive (CK+) cells in the bone marrow (BM) and serum tumour markers (TM) in patients with non small cell lung cancer (NSCLC) at the time of diagnosis with regard to overall survival (OS) and time to progression (TTP). Eighty patients with NSCLC, staged as T2-4, N0-3, M0 (n=52), M1 (n=27), (Mx = 1) were evaluated. Treatment included chemo-radiotherapy with cisplatin/etoposide and subsequent radical surgical resection. There were 23 complete responders, 50 non-responders and 7 patients who died of non-lung cancer causes. The median follow-up was 12 months (range 1 44 months). Besides routine pathology for tissue and BM, CK+ BM cells were detected by immunocytochemistry (IC) and 4 different tumour markers as well as the shedded domain of the oncoprotein Her-2/neu and urokinase plasminogen activator uPA were determined by radio- or enzyme-immunoassay. Patients classified as stage IV and patients with metastases had a significantly lower TTP and OS. No significant correlation was demonstrated for grading, tumour size or number of involved lymph nodes. The tumour marker tissue polypeptide antigen (TPA) and Cyfra 21-1 were the only marker which significantly correlated with OS. Interestingly, routine pathology could not detect minimal residual BM involvement as IC was able to (p=0.0004) and the presence of even a few CK+ cells significantly correlated with reduced OS. Thus, we conclude that the detection of CK+ cells should be added to routine pathology and for tumour marker determination, studies should focus on Cyfra 21-1 and TPA. PMID- 12579293 TI - Glutathione S-transferase M1 associated with cancer occurrence in Korean HNPCC families carrying the hMLH1/hMSH2 mutation. AB - Hereditary non-polyposis colorectal cancer (HNPCC), an inherited cancer predisposition syndrome, has been associated with germline mutations in DNA mismatch repair (MMR) genes. However, because all mutation carriers of hMLH1/hMSH2 do not account for CRC susceptibility, modifying genes may play a role in the variation of disease expression. In this study, we determined the GSTM1 and GSTT1 genotypes in 104 family members representing 19 Korean HNPCCs carrying hMLH1/hMSH2 mutation, and investigated the influence of GSTM1 and GSTT1 geno-/phenotype status on both age at diagnosis of CRC and cancer occurrence. The overall frequency of the GSTM1 and GSTT1 geno-/phenotype in 55 non-carriers, compared with that in mutation carriers (n=49), was not significantly different, and no significant correlation was found between mean age at diagnosis and the allelomorphs encoding the GSTM1 or GSTT1 enzymes. However, a comparison of the allele frequencies of GSTM1 in affected (n=30) and unaffected (n=19) mutation carriers revealed a significant difference, as the null allele was more prevalent in individuals with cancer (p=0.03; odds ratio, 3.7; 95% confidence interval, 1.1 12.7). Our results suggest that the genotypes of GSTM1 are associated with cancer occurrence in Korean HNPCC family members carrying the hMLH1/hMSH2 mutation. However, a bias due to the small sample size of this study cannot be rule out. Although evidence that GST genotypes are associated with increased cancer risk has often been controversial, the genotyping of GSTM1 could have implications for genetic counseling and the management of MMR gene mutation carriers. PMID- 12579294 TI - Low frequency of human polyomavirus BKV and JCV DNA in urothelial carcinomas of the renal pelvis and renal cell carcinomas. AB - Human polyomaviruses BKV and JCV establish a persistent infection in the kidney and possess oncogenic potential in experimental animal models. Their possible involvement in the pathogenesis of human urogenital tumors has been suggested by several studies but is not conclusively resolved. For the present study, highly sensitive quantitative TaqMan PCR assays were developed to detect BKV and JCV DNA sequences in matched pairs of human tumors and surrounding normal tissue. Samples from 55 patients with renal pelvic urothelial carcinomas and from 83 patients with renal cell carcinomas of all histologic subtypes were investigated. Human polyomavirus DNA was detected in renal pelvis samples with a frequency of 16% (BKV) and 15% (JCV), and in kidney samples with a frequency of 1% (BKV) and 23% (JCV), respectively. However, viral sequences were not present more often in tumors than in normal tissue. There was no integration of the potentially oncogenic viral large T-antigen in the tumors, and the viral concentration of both BKV and JCV was not higher in DNA extracted from tumors compared to normal tissue. These data provide no evidence for a causative role of the human polyomaviruses BKV and JCV in the development of tumors of the renal pelvis and the kidney. PMID- 12579295 TI - Meizothrombin, an intermediate of prothrombin cleavage potently activates renal carcinoma cells by interaction with PAR-type thrombin receptors. AB - Recently, meizothrombin (MT), an intermediate enzyme in the prothrombin cleavage cascade has been shown to activate cells of a brain tumor cell line by interaction with PAR-1-type thrombin receptors with a potency comparable to that of thrombin. In this study, we investigated the effect of recombinant human MT (rMT) on calcium mobilization in primary cultures established from surgically resected human renal cell carcinomas. Meizothrombin induced very rapidly transient calcium mobilization in RCC cells comparable to that observed with thrombin. RCC cells stimulated with thrombin after rMT challenge were unable to elicit a new calcium response and vice versa. Therefore, rMT and thrombin seem to activate calcium signaling in primary RCC cultures by similar mechanisms including PAR-1- and PAR-3-type thrombin receptors as shown by using PAR-type specific antibodies. Our results demonstrate rMT as a potent activator of human RCC cells suggesting a function of not only thrombin but also of this catalytically active thrombin precursor enzyme in human renal cell carcinoma. PMID- 12579296 TI - Beta-sitosterol, a plant sterol, induces apoptosis and activates key caspases in MDA-MB-231 human breast cancer cells. AB - The objective of the present study was to evaluate the effect of beta-sitosterol, a plant sterol that induces apoptosis in breast cancer cells, on two pathways leading to apoptosis. These pathways are classified based on the localization of the initiated signal, extrinsic and intrinsic pathways. Extrinsic and intrinsic pathways are catalyzed by caspases 8 and 9, respectively, which leads to the activation of the executioner caspase 3. The results of the present study indicate that beta-sitosterol supplementation at 16 microM for 3 days to MDA-MB 231 cells induces 39% and 80% increases in the activities of caspases 8 and 9, respectively, compared to cholesterol supplemented cells or controls. There was also a 3-fold increase in the activity of caspase 3. Sterol treatment had no effect on the quantities of the enzymes. It is concluded that beta-sitosterol may induce apoptosis through the two pathways but was more pronounced on the intrinsic pathway. PMID- 12579297 TI - Epigenetic control of E-cadherin (CDH1) by CpG methylation in metastasising laryngeal cancer. AB - Intercellular adhesion is promoted by many-fold structures formed by interacting molecules. One prominent protein family, called cadherins, consists of calcium dependent proteins contributing to cell differentiation, migration and extracellular signal transduction. E-cadherin, regularly expressed in epithelial tissues, displays aberrant activity patterns in a variety of tumors. We have explored the mode of E-cadherin regulation in 98 biopsy samples from 76 patients with laryngeal carcinoma (80 primary tumors and 18 metastases). Transcriptional silencing of the gene (CDH1) achieved by promoter methylation was tested by a methylation-specific PCR. In primary tumors, CDH1 methylation was noted in 40%. However, 77% of the tested metastases showed CDH1 methylation, 23% remained unmethylated (p<0.01). These data suggest that the E-cadherin promoter is subjected to epigenetic control connected with biological aggressiveness of laryngeal cancer in advanced steps of neoplastic transformation. PMID- 12579298 TI - Ultrasonographic detection of regional lymph node metastases in patients with intermediate or thick malignant melanoma. AB - Careful monitoring of regional lymph nodes and early detection of metastases in malignant melanoma patients has an impact on their survival, since it may permit beneficial surgical therapy. Palpation is routinely used in clinical practice. The value of ultrasonography for routine follow-up of melanoma patients, still, is not generally accepted. The aim of our study was to assess the sensitivity and specificity of ultrasound and clinical examination respectively, in the detection of melanoma regional node metastases. Additionally, we evaluated whether early detection of metastases improved overall survival. One hundred and forty-eight melanoma patients with an intermediate or thick primary lesion were followed between January 1997 and May 2001. Clinical examination and concomitant regional lymph node ultrasonography were performed, every 3-4 months. If suspicious findings were identified, regional lymph node dissection was undertaken. Forty four from the initial 148 patients relapsed with regional lymph nodal metastases. In 11 patients (25%) palpation failed to reveal the disease and metastases were depicted only by ultrasonography. In only 1 patient ultrasonography was false negative. The sensitivity and specificity of palpation were 72.7 and 97% respectively, while those of ultrasonography were 97.7 (p<0.001) and 98% respectively. Ultrasonography was more sensitive in detecting lymph node metastases in the axilla (100%) and the groin (93.3%). When overall survival of patients presenting with local-regional recurrence was calculated--depending on the number of involved lymph nodes--a survival benefit (p<0.05) was found for patients with only one lymph node metastasis. In conclusion, ultrasonography is superior to clinical examination in the early detection of regional lymph node metastases from an intermediate or thick malignant melanoma and should be a part of those patients' surveillance. PMID- 12579300 TI - CD26: a novel treatment target for T-cell lymphoid malignancies? (Review). AB - CD26 is a surface glycoprotein with intrinsic dipeptidyl peptidase IV (DPPIV) enzyme activity with multiple biological roles, including being intricately involved in immunoregulation as a T-cell activation molecule and as a regulator of chemokine function. T-cell lymphoid malignancies represent a heterogeneous group of diseases that are generally aggressive and are for the most part resistant to current treatment modalities. Previous studies showed that CD26 is expressed on selected T-cell neoplasms, suggesting a potential role for CD26 in tumor development. We review herein recent classification schemes for T-cell lymphoid malignancies that take into account various facets of their clinical presentation. In addition, we discuss findings supporting the conclusion that CD26 has an essential role in human T-cell activation, as well as its ability to regulate the biological effects of selected chemokines through its DPPIV activity. Finally, we will present recent work from our laboratory that indicates a potential role for CD26 as a molecular target for novel treatment modalities for T-cell lymphoid malignancies. PMID- 12579299 TI - Regulation of cell cycle progression and apoptosis by the Ras/Raf/MEK/ERK pathway (Review). AB - The Ras/Raf/MEK/ERK signal transduction pathway regulates cell cycle progression and apoptosis in diverse types of cells. Mutations in this pathway are often observed in transformed cell lines and frequently linked with human cancers. The Ras/Raf/MEK/ERK pathway can induce events both associated with cell proliferation and cell cycle arrest. The particular course chosen may depend on the strength and the particular Raf gene activated by Ras. This pathway also is involved in maintaining cell survival by modulating the activity of apoptotic molecules including Bad and Bcl-2. This review will discuss the regulation of the Ras/Raf/MEK/ERK pathway and how it modulates cell cycle progression and cell survival. PMID- 12579301 TI - Medical hypothesis: hyperhomocysteinemia is a risk factor for estrogen-induced hormonal cancer. AB - A novel mechanistic hypothesis is proposed which suggests that hyperhomocysteinemia is a risk factor for the development of estrogen-induced hormonal cancer in humans. Mechanistically, hyperhomocysteinemia may exert its pathogenic effects largely through metabolic accumulation of intracellular S adenosyl-L-homocysteine, a strong non-competitive inhibitor of the catechol-O methyltransferase-mediated methylation metabolism of endogenous and exogenous catechol estrogens (mainly 2-hydroxyestradiol and 4-hydroxyestradiol). While a strong inhibition of the methylation metabolism of 2-hydroxyestradiol would decrease the formation of 2-methoxyestradiol (an antitumorigenic endogenous metabolite of 17beta-estradiol), an inhibition of the methylation of 4 hydroxyestradiol would lead to accumulation of this hormonally-active and strongly procarcinogenic catechol estrogen metabolite. Both of these effects resulting from inhibition of the methylation metabolism of catechol estrogens would facilitate the development of estrogen-induced hormonal cancer in the target organs. This hypothesis also predicts that adequate dietary intake of folate, vitamin B6, and vitamin B12 may reduce hyperhomocysteinemia-associated risk for hormonal cancer. Experimental studies are warranted to determine the relations of hyperhomocysteinemia with the altered circulating or tissue levels of 4-hydroxyestradiol and 2-methoxyestradiol and also with the altered risk for estrogen-induced hormonal cancer. PMID- 12579302 TI - Novel cell culture models for prevention of human breast cancer (Review). AB - Human breast cancer is a multifactorial, multistep disease wherein genetic, endocrine and dietary factors represent crucial regulators of initiation, promotion and progression. Preclinical investigations utilizing human breast carcinoma derived cell lines either in culture, or upon xenotransplantation, have provided valuable leads for molecular pathogenesis of cancer progression and also for novel therapeutic modalities. The mechanistic significance of genetic factors on early events of initiation/promotion, however, is dependent on extrapolation, and is therefore, equivocal. Human tissue derived explant culture/cell culture models utilizing non-involved target tissue at risk for carcinogenic transformation provide a novel approach that minimizes extrapolation for clinical relevance and thereby maximizes the translational impact. This report provides an overview of laboratory investigations focused on: i) development of the model, ii) optimization of mechanistic biomarker assays for carcinogenic transformation, and iii) validation of the model as a high throughput mechanistic screen for preclinical efficacy of natural phytochemicals. PMID- 12579303 TI - Atypical nuclear apoptosis downstream to caspase-3 activation in ara-C treated CCRF-CEM cells. AB - The necessity of internucleosomal DNA fragmentation for the complete execution of apoptosis is still controversial. While investigating the apoptotic pathway induced by 1-beta-D-arabinofuranosylcytosine (ara-C) in the human T-lymphoblastic leukemia CCRF-CEM (CEM) cells, we could easily retrieve high molecular weight (HMW) DNA fragments with a predominant size of 50 kb. However, under the same circumstances, internucleosomal DNA fragmentation characteristic of apoptosis was undetectable despite estimated heightened caspase-3 activity. Paradoxically, generation of low molecular weight DNA fragments was readily demonstrable by flow cytometric and immunohistochemical evidence in the absence of any detectable DNA ladder formation. These findings present a proof that, within certain contexts, small-sized DNA fragmentation occurring in apoptosis may not necessarily be of the ladder yielding internucleosomal integer multiples' pattern. Our data also add to the evidence that the machinery underlying HMW DNA fragmentation is distinct from that responsible for the internucleosomal one. PMID- 12579304 TI - Fluorescence confocal microscopy and image analysis of bladder cancer using 5 aminolevulinic acid. AB - 5-aminolevulinic acid mediated changes in tissue specific fluorescence were studied in bladder cancer. Bladders of normal patients and also patients diagnosed with cancer were instilled with 5-aminolevulinic acid and the resultant protoporphyrin IX mediated fluorescence intensity was imaged and quantified with confocal laser microscopy and fluorescence image analysis. Urothelial tumour cells were observed to fluoresce more intensely than normal urothelial cells. Submucosa and muscle tissues exhibited minimal fluorescence compared to urothelial cells of malignant origin and also normal urothelial cells. Degree of fluorescence intensity was in the order of malignant urothelium > normal urothelium > normal submucosa > normal muscles. Fluorescence intensity was also found to increase with duration of ALA instillation. Grade 3 malignant cells produced more fluorescence compared to grade 2 and grade 1. Similarly, T1 transitional cell carcinoma (TCC) showed increased fluorescence intensity than that of Ta TCC. Also, tumour blood vessels fluoresced more intensely compared to blood vessels found in normal bladder tissue. Tissue specific ALA mediated PpIX micro fluorescence can be used as a diagnostic technique for early detection of neoplasms and confocal laser microscopy and fluorescence image analysis are advantageous diagnostic tools for the photodynamic diagnosis of bladder neoplasms in vivo. PMID- 12579305 TI - Reduced expression of heterogeneous nuclear ribonucleoprotein B1 in adult T-cell lymphoma/leukemia. AB - It is considered that hnRNP B1 expresses similarly in the various types of tumor cells. Recently, we demonstrated high B1 expression in B-cell lymphoma and carcinoma. To evaluate the difference of B1 expression between B and T-cell lymphoma, we immunologically studied the B1 expression in 22 cases with nodal T cell lymphoma, comprising adult T-cell leukemia/lymphoma (ATLL; n=15) and angioimmunoblastic T-cell lymphoma (AILD; n=7), using an anti-hnRNP B1 monoclonal antibody, 2B2. In ATLL cases, scattered large transformed lymphoma cells demonstrated strong B1 expression, while the medium-sized lymphoma cells were negative. On the one hand, lymphoma cells in AILD diffusely expressed B1. The mean B1 expression rate in ATLL was 22%, which was significantly lower than that in AILDs (45%), B-cell lymphomas (44%), and metastatic carcinomas (53%) (p<0.01). Our result might suggest that process of hnRNP B1 expression in ATLL differs from those in other lymphoid neoplasms and carcinoma. PMID- 12579306 TI - Establishment of a novel human dedifferentiated liposarcoma cell line, FU-DDLS-1: conventional and molecular cytogenetic characterization. AB - A number of human cell lines derived from well-differentiated, myxoid/round cell, or pleomorphic liposarcoma have been described. To our knowledge, however, no human cell line established from dedifferentiated liposarcoma has been reported. In this study, we established a new human cell line, FU-DDLS-1, which originated from a dedifferentiated liposarcoma arising in the retroperitoneum of a 61-year old man. This cell line was characterized by immunocytochemistry, conventional banding analysis, fluorescence in situ hybridization with chromosome painting probe, and comparative genomic hybridization (CGH). FU-DDLS-1 cells were spindle or polygonal shaped and possessed oval nuclei and slender cytoplasmic processes. The cultured cells were successfully maintained in vitro for over 90 passages over more than 30 months. The histologic features of heterotransplanted tumors in severe combined immunodeficiency mice were essentially the same as those of the original nonlipogenic sarcoma resembling a malignant fibrous histiocytoma. Both in vitro and in vivo, the cells exhibited immunopositive reaction for mdm2 and p53 proteins. Cytogenetically, FU-DDLS-1 displayed a hypertetraploid karyotype with giant marker chromosomes composed partly of chromosome 12 material. In addition, CGH analysis demonstrated that DNA sequence copy number changes including a gain of 12q12-q21 detected in FU-DDLS-1 were essentially the same as those in the original sarcoma. The FU-DDLS-1 cell line, which exhibits the unique conventional and molecular cytogenetic characteristics of dedifferentiated liposarcoma, should be a particularly useful model for studying the molecular pathogenesis of human dedifferentiated liposarcoma. PMID- 12579307 TI - In situ detection of O6-methylguanine-DNA methyltransferase messenger RNA in paraffin-embedded human astrocytic tumor tissues by nested in situ RT-PCR is useful in predicting chemotherapy-resistance of tumors. AB - O6-methylguanine-DNA methyltransferase (MGMT) is an enzyme that interferes with chemotherapeutic effect of alkylating agents. We performed in situ detection of MGMT mRNA utilizing the nested RT-PCR method in tissue sections (nested in situ RT-PCR). We analyzed 34 samples of paraffin-embedded astrocytic tumor tissue sections with this method [3 astrocytomas, 14 anaplastic astrocytomas (AA), and 17 glioblastoma multiformes (GBM)]. Twenty-five cases (73.5% of all cases) were positive for MGMT either with our method or immunohistochemistry (IHC). Moreover, with our method >25% of the cells in the tumor tissue expressed MGMT in contrast to >4% with IHC among the population of MGMT positive cases. Our method was significantly more sensitive than IHC (p=0.0004). The present results suggest that potentially there is a greater population of MGMT positive cells in astrocytic tumor tissues than the one evaluated with IHC. These findings suggest that the >25% of the MGMT positive cells are involved in the interference with the chemotherapeutic effect of alkylating agents. The MGMT expressing cell population was markedly decreased in GBM compared with AA (26.1% vs 62.1%). The main reason for this marked decrease was that MGMT was expressed in only 9 of 17 cases of GBM in contrast to all AA cases that expressed MGMT. This result suggests that there are potentially two populations of GBM on the basis of MGMT expression, in which the negative population might be mainly composed of de novo GBM. Therefore, it is suggested that our method is practically useful to detect any drug resistance gene product with high sensitivity and would provide a chance to evaluate the chemotherapeutic effect of any agents in an individual patient based manner. PMID- 12579308 TI - Genome-wide cDNA microarray analysis of gene-expression profiles involved in ovarian endometriosis. AB - Using a cDNA microarray consisting of 23,040 genes, we analyzed gene-expression profiles of ovarian endometrial cysts from 23 patients in order to identify genes involved in endometriosis. By comparing expression patterns between endometriotic tissues and corresponding eutopic endometria, we identified 15 genes that were commonly up-regulated in the endometrial cysts during both proliferative and secretory phases of the menstrual cycle, 42 that were up-regulated only in the proliferative phase, and 40 that were up-regulated only in the secretory phase. The up-regulated elements included genes encoding some HLA antigens, complement factors, ribosomal proteins, and TGFBI. On the other hand, 337 genes were commonly down-regulated throughout the menstrual cycle, 144 only in the proliferative phase, and 835 only in the secretory phase. The down-regulated elements included the tumor suppressor TP53, genes related to apoptosis such as GADD34, GADD45A, GADD45B and PIG11, and the gene encoding OVGP1, a protein involved in maintenance of early pregnancy. Semi-quantitative RT-PCR experiments supported the results of our microarray analysis. These data should provide useful information for finding candidate genes whose products might serve as molecular targets for diagnosis or treatment of endometriosis. PMID- 12579309 TI - Methylation of promoter region in p27 gene plays a role in the development of lymphoid malignancies. AB - p27 belong to the cyclin-dependent kinase inhibitor family and plays an important role in regulation of cell cycle progression. Expression of these genes is epigenetically suppressed by methylation of their promoter regions. In this study, we examined protein expression and methylation status of the promoter region of p27 in 70 cases with non-Hodgkin's lymphoma and their relationship with proliferative activity of the tumor cells as revealed by Ki-67 index. There were 35 cases of B-cell, 11 of T-cell, 23 of NK/T-cell lymphoma, and 1 undetermined type. Immunohistochemically the loss of p27 protein expression was observed in 24 (69%) of 35 cases. Significant inverse correlation between p27 protein expression and Ki-67 index was found. Single strand conformation polymorphism (SSCP) followed by sequencing could not detect point mutations in any of the 68 cases. Bisulfite sequencing analysis showed that methylation of the promoter region of p27 were observed in 17 (25%) of 68 cases, in that several specific CpG sites around the start codon of p27 gene were preferentially methylated. Eight of 22 cases with loss of p27 protein expression showed methylation in CpG island of 5' region of p27 gene. These findings suggested that gene methylation plays a role in loss of expression of p27, which gives the cells proliferative advantages. PMID- 12579310 TI - Combined suicide gene and immunostimulatory gene therapy using AAV-mediated gene transfer to HPV-16 transformed mouse cell: decrease of oncogenicity and induction of protection. AB - To test the effects of combined transduction of a suicide gene and genes coding for various immunostimulatory factors on the oncogenicity and immunogenicity of TC-1 cells (HPV-16 transformed C57BL/6 mouse cells), several bicistronic recombinant adeno-associated viruses (rAAV) were constructed. Each of these constructs carried, and in infected cells expressed, the herpes simplex type 1 thymidine-kinase gene (HSV-TK) and the gene of one of the following immunostimulatory factors: human monocyte chemoattractant protein 1 (MCP-1), mouse B7.1 costimulatory molecule (B7.1), or mouse granulocyte-macrophage colony stimulating factor (GM-CSF). For control purposes, an rAAV carrying the HSV-TK gene and neomycin resistance gene (neo) and an rAAV containing the lacZ gene were used. All of these constructs proved functional both in mouse TC-1 and human 293T cells. For experiments in mice, TC-1 cells were infected in vitro with the AAV recombinants at an input multiplicity of 50 particles/cell; these cells were then administered to 5-week-old mice. As from day 5, half of the animals were given ganciclovir (GCV) (2.5 mg/day) for 10 days. With a single exception, none of the mice inoculated with cells treated with rAAV expressing HSV-TK + B7.1 or HSV-TK + MCP-1 developed tumour irrespective of GCV treatment. The tumour suppressive effect was less marked in animals inoculated with TC-1 cells infected with rAAV expressing HSV-TK + GM-CSF, and among these it was somewhat more pronounced in GCV-untreated animals. A clear antitumour effect of GCV treatment was only observed in mice inoculated with TC-1 cells transduced with rAAV expressing HSV TK but no immunostimulatory factor. Mice that remained tumour-free on day 54 were challenged with untreated TC-1 cells. The tumour resistance rates found were related not only to the immunostimulatory gene used for the transduction, but also to GCV treatment. The best protection was recorded in mice pre-inoculated with TC-1 cells transduced with either B7.1 or MCP-1-expressing rAAV and not given GCV. PMID- 12579311 TI - Apoptosis and tumor remission in liver tumor xenografts by 4-phenylbutyrate. AB - 4-phenylbutyrate (triButyrate trade mark, PB) a derivative of the short-chain fatty acid, butyrate, possesses anti-tumor activity in vitro in different tumor cell lines. Unlike most cytostatic compounds, PB possesses low toxicity. In order to evaluate possible clinical use of PB in cancer therapy, hepatocarcinoma (Hep3B) and hepatoblastoma (HepT1) cell lines, as well as xenografts derived from those in nude rats, were treated with PB in different dose (1-100 mM) and time regimens. Treatment with 10 mM of PB for 24 h (or 5 mM for 48 h) was shown to significantly inhibit Hep3B cell growth in vitro. The HepT1 cell line was more sensitive to PB treatment: already 1 mM of PB for 24 h significantly inhibited the growth of the cells. PB also resulted in regression of xenografts derived from these cell lines in vivo, when administrated by mini-pump with an intratumor catheter, yielding 20 micro mol of PB per cm3 of tumor volume per day. TUNEL assay and caspase-3 activity measurements suggested apoptosis to be the cell death mechanism in both cell lines and xenografts. Increased histones H3 and H4 acetylation was shown in both cells and xenografts, and the inhibition of histone deacetylase is proposed as the main trigger for the anti-tumor action of PB. Concomitant induction of p21Waf1/Cip1 expression was detected by RNase protection assay and Western blotting. Reduction in expression of alpha-fetoprotein was found both in Hep3B cells and xenografts, suggesting also a differentiation effect by PB. PMID- 12579312 TI - In-depth analysis of molecular alterations within normal and tumour tissue from an entire bronchial tree. AB - Using laser capture microdissection (LCM), fluorescent microsatellite analysis and immunohistochemical analysis, we have constructed a detailed topographical molecular map of the entire bronchial tree surrounding a primary bronchial squamous carcinoma in order to establish the relationship between the molecular damage within the airway and that in the tumour itself. Allelic imbalance was analysed using markers on chromosomes 3, 9, 13 and 17. In addition, immunohistochemical analysis for p53 and cyclin D1 expression was performed. Analysis revealed allelic imbalance at several loci at the tumour site but also in 83% of the histologically normal airway specimens of the upper and lower lobes. The fractional allele loss (FAL) value was statistically higher (0.75+/ 0.13) in the tumour site than in the distal site of the upper (0.42+/-0.09) and lower lobes (0.31+/-0.08). Immunohistochemical analysis revealed overexpression of p53 and cyclin D1 protein within histologically normal bronchial epithelium, thus confirming previous reports for their early involvement in lung tumour development. This is to date the largest in-depth study of allelic imbalance using LCM in a single individual. The patterns of allele-specific imbalance observed support a clonal or oligoclonal expansion model of outgrowths throughout the lung. The widespread incidence of genetic changes in the whole of lung most likely represents smoking-induced alterations and emphasize the complexity of the field cancerization concept. Our findings point to the need for in-depth studies of the whole bronchial tree tissue surrounding lung carcinomas, in order to identify the genetic changes that differentiate preneoplastic and neoplastic stages in lung carcinogenesis. PMID- 12579313 TI - Changes in mitochondrial membrane potential and mitochondrial mass occur independent of the activation of caspase-8 and caspase-3 during CD95-mediated apoptosis in peripheral blood T cells. AB - Although death receptor pathway and mitochondrial pathways of apoptosis are connected through Bid cleavage, such activation of mitochondrial pathway by death receptor signaling is observed in type II cells and not in type I cells (peripheral blood T cells). Furthermore, activation of mitochondria via Bid is associated with release of cytochrome c and caspase-9 activation. In this study we demonstrate that anti-CD95-induced apoptosis in T cells is associated with both depolarization of mitochondrial membrane potential (Delta(psi)m) and increase in mitochondrial mass and activation of caspase-8 and caspase-3 but without caspase-9 activation. Furthermore, changes in mitochondrial membrane potential and mitochondrial mass by anti-CD95 monoclonal antibodies were unaffected by inhibitors of caspase-8 and caspase-3, suggesting that anti-CD95 induced changes in Delta(psi)m and mitochondrial mass are independent of caspase 8 and caspase-3 activation. PMID- 12579314 TI - Promoter hypermethylation of multiple genes in astrocytic gliomas. AB - Promoter hypermethylation represents a primary mechanism in the inactivation of tumor suppressor genes during tumorigenesis. To determine the frequency and timing of hypermethylation during carcinogenesis of astrocytic tumors, we analysed promoter methylation status of ten tumor-associated genes (MGMT, GSTP1, DAPK, p14ARF, THBS1, TIMP-3, p73, p16INK4A, RB1 and TP53) in a series of 88 astrocytic gliomas, including 24 diffuse astrocytomas; 21 anaplastic astrocytomas, and 43 glioblastomas (33 primary and 10 secondary), as well as two non-neoplastic brain samples, by methylation-specific PCR. Aberrant CpG island methylation was detected in all ten genes analysed, and all but one sample displayed anomalies in at least one gene. The methylation index (number methylated genes/total genes analysed) was 0.3, 0.38, 0.33 and 0.29 for diffuse astrocytomas, anaplastic astrocytomas and secondary and primary glioblastomas, respectively. Some differences may be established regarding the methylation profiles of specific genes and tumor types: MGMT, THBS1, TIMP-3, and p16INK4A appear hypermethylated in low-grade tumors (at least in 45% of cases), whereas GSTP1, DAPK, and p14ARF are mostly changed in 15-50% of the higher grade forms versus <10% in low-grade tumors. Some variation also exists regarding the methylation values for p73 and RB1 (10-40% of cases) among all groups. TP53 presented hypermethylation rates <10% in all tumor subtypes. Our findings thus suggest that methylation represents a common mechanism that contributes to inactivating cancer-related genes in astrocytic neoplasms. This epigenetic change is, in general, an early event in the development of astrocytic neoplasms but this gene silencing mechanism may also appear as a late event involving some loci. PMID- 12579315 TI - Estradiol-induced vascular endothelial growth factor-A expression in breast tumor cells is biphasic and regulated by estrogen receptor-alpha dependent pathway. AB - Estrogens have been shown to regulate vascular endothelial growth factor-A (VEGF A) for physiological and patho-physiological functions. However, estrogen action on VEGF-A mRNA expression has not been completely elucidated. We have identified two phases of activation of VEGF-A mRNA transcription, one early and one late response, induced by 17beta-estradiol (17beta-E2) in ER+ MCF-7 breast tumor cells, depending upon the length of exposure. VEGF-A mRNA level was significantly higher than control in tumor cells after 2 h of 17beta-E2 exposure. Furthermore this induction was not inhibited by cycloheximide, indicating that it was a direct effect of estrogen. In contrast VEGF-A mRNA expression was back at basal level in MCF-7 cells exposed to 17beta-E2 for 6 h. However, expression levels were again significantly augmented after 24 h of exposure, and this induction was unaltered by cycloheximide indicating that de novo protein synthesis was not required and like early response, it was a direct effect of estrogen. The antiestrogen ICI 182,780 was a pure antagonist for the early response phase of VEGF-A mRNA induction, but it had partial but significant effect on the late response phase, further suggesting that both early and late phases were ER dependent. In human mammary epithelial cells (HMEC) lacking estrogen receptor (ER alpha) the early and late response phase of VEGF-A mRNA induction in response to 17beta-E2 was not found, but significant inductions were seen in the early and late phases when ER-alpha transfected HMEC were exposed to 17beta-E2 for 2 or 24 h. Taken together, these studies suggest that VEGF-A is an estrogen responsive gene and modulation of this gene expression by estrogen is biphasic and can be mediated through ER-alpha dependent pathway. PMID- 12579316 TI - High-resolution profiling of an 11 Mb segment of human chromosome 22 in sporadic schwannoma using array-CGH. AB - Previous low-resolution schwannoma studies have reported diverse frequencies (30 80%) of 22q deletions, involving the neurofibromatosis-2 tumor suppressor (NF2) gene. We constructed an array spanning 11 million base pairs of 22q encompassing the NF2 gene, with 100% coverage and an average resolution of 58 kb. Moreover, the 220 kb genomic sequence encompassing the NF2 gene was covered by 13 cosmids to further enhance the resolution of analysis. The rationale of this array-CGH study was to map and size 22q deletions around the NF2 gene in sporadic schwannoma using a reliable method with maximal resolution. We studied tumor and constitutional DNA from 47 patients and detected heterozygous deletions in 21 (45%) tumors, which could be classified into three profiles. The predominant profile (12/21) was a continuous deletion of the 11 Mb segment, consistent with monosomy 22. The second profile, comprising five schwannomas, was also in agreement with a continuous 11 Mb heterozygous deletion. However, these displayed a distinctly different level of deletion when compared to the first profile, suggesting a considerable amount of normal tissue in the tumor samples. This is the first report demonstrating the sensitivity of array-CGH to discriminate such samples. The third profile was composed of four cases displaying interstitial deletions of various sizes. Two of these did not encompass the NF2 locus, which further emphasize the importance of other loci in schwannoma development. This is the first high-resolution study performed on a large series of tumors, using an array continuously covering 1/3 of a human chromosome. Our findings warrant further studies of an extended tumor series on a full 22q genomic array, to better define additional, putative 22q-located loci important for schwannoma development. Our array also provides a new diagnostic tool for analysis of NF2 gene deletions in patients affected with neurofibromatosis-2. PMID- 12579317 TI - Differential effects of retinoic acid on the growth of isogenic metastatic and non-metastatic breast cancer cell lines and their association with distinct expression of retinoic acid receptor beta isoforms 2 and 4. AB - The human retinoic acid receptor beta (RARbeta) has three isoforms (beta1, beta2, and beta4), which play important, distinct roles in mediating the effects of retinoic acid on cell growth and apoptosis. Whereas RARbeta2 is a potent inhibitor of breast cancer cell proliferation, RARbeta4 can act as a dominant negative repressor of RARbeta2-mediated growth suppression. In this study we investigated the effects of all-trans-retinoic acid (ATRA) on two clones derived from the breast cancer cell line MDA-MB-435: a non-metastatic clone (NM-2C5) and a metastatic clone (M-4A4). ATRA treatment of the NM-2C5 cells resulted in growth inhibition and apoptosis, whereas the M-4A4 cells were resistant to ATRA. Analyses of the expression of RARbeta isoforms revealed that the sensitive NM-2C5 clone expressed only RARbeta2, whereas the resistant M-4A4 cells expressed both RARbeta2 and RARbeta4 mRNA and protein. ATRA treatment increased RARbeta2 mRNA level in NM-2C5 cells, whereas the same treatment of the M-4A4 cells resulted in an increase in RARbeta4 and a decrease in RARbeta2 mRNA. ATRA treatment of NM-2C5 cells increased the protein levels of the histone acetyl transferases p300 and CBP, suppressed the level of histone deacetylase and increased the level of acetylated histone H4. ATRA also decreased Bcl-2 and increased Bax and decreased VEGF. In contrast, the same treatment of the M-4A4 cells resulted in opposite effects. These results suggest that the effects of ATRA on the growth of the metastatic and non-metastatic breast cancer cell lines depend on the expression of RARbeta isoforms and that the expression of RARbeta4 may contribute to metastatic properties. PMID- 12579318 TI - Requirement of soluble factors produced by bone marrow stromal cells on the growth of novel established human myeloma cell line. AB - The growth of myeloma cells is believed to be mediated by functional interactions between tumor cells and the marrow environment involving the action of several cytokines. We report on the establishment and characterization of a new human myeloma cell line (TAB1) that can be long-term maintained in the presence of conditioned medium of bone marrow stromal cells (BMCM) and a BMCM independent variant, C2-2. Both cell lines have plasma cell morphology and express plasma cell antigens (CD38, PCA-1 and immunoglobulin kappa light chain). In the absence of BMCM, TAB1 cells undergoing apoptosis were observed. Among the adherent molecules tested, these cells expressed VLA-4, ICAM-1 and H-CAM, but not VLA-5, suggesting that these were mostly immature plasmacytes. Introduction with exogenous IL-6 and/or GM-CSF, which were detected in BMCM, partially supported the proliferation of TAB1 cells. Treatment with anti-IL-6 antibody partially inhibited the proliferation of TAB1 cells cultured with BMCM. These findings strongly suggest that TAB1 required at least two or more factors on their growth in vitro; IL-6 was one of the factors necessary for cell growth. Further studies are required to clarify the precise molecules which support TAB1 cell growth in combination with IL-6, however, TAB1 and its variant C2-2 cells may offer an attractive model to unravel novel molecular mechanisms involved in bone marrow stroma-dependent growth of myeloma cells. PMID- 12579319 TI - The usefulness of 99mTc-tetrofosmin SPECT in the detection of intrathoracic malignant lesions. AB - To evaluate the usefulness of 99mTc-tetrofosmin (TF) SPECT in the detection of intrathoracic malignant lesions, we studied 304 patients, 261 with malignant and 43 with benign lesions; 196 of the former had non-treated primary tumors, 193 lung cancer (LC) and 3 mesotheliomas, 11 had LC recurrences and 54 had metastases from different kinds of tumors. Twenty-nine patients with primary tumors were re checked after chemotherapy or surgery. In all patients, after 740 MBq TF injection, both planar and SPECT images were acquired and analysed qualitatively, SPECT images also semiquantitatively. Scintigraphy was always compared to CT. SPECT showed higher sensitivity, specificity and accuracy values than CT (98.1, 90.7 and 97 vs. 96.2, 81.4, and 93.9%, respectively), their combined use achieving 100% sensitivity and 98.8% accuracy. Planar did not give more information than SPECT, showing a significantly lower sensitivity (63.2%) and accuracy (67.4%). SPECT showed higher accuracy values than CT and planar (86.9 vs. 78.3 and 69.6%) in NSCLC mediastinal lymph node staging. Moreover, SPECT was concordant with CT in correctly evaluating the response to chemotherapy or surgery in all monitorized primary tumors cases, except in one in whom only SPECT detected residual tumor. The semiquantitative analysis added useful information in differentiating malignant from benign lesions and in monitoring the response to chemo-therapy. TF SPECT appears a highly accurate diagnostic method in the detection of intrathoracic malignant lesions, in lungs and pleura, as well as in NSCLC mediastinal lymph node staging and in monitoring treatment effectiveness, playing a complementary role to CT in selected cases. PMID- 12579320 TI - In vivo induction of dendritic cell-mediated cytotoxicity against allogeneic pancreatic carcinoma cells. AB - The objective of this study was to assess the toxicity and immunological response induced by autologous dendritic cells (DCs) pulsed with allogeneic tumor lysate in a pancreatic cancer patient. The lack of available tumor peptides in pancreatic cancer strongly supports the idea to use allogeneic tumor cells as a source of antigens. The patient suffering from a stage IV pancreatic cancer received 1-2x10(7) autologous monocyte-derived DCs in three-week intervals injected into a groin lymph node. Monocytes from peripheral blood were isolated by magnetic bead selection. For the first ten vaccinations DCs were loaded with autologous tumor cell lysate obtained during surgical exploration. After consumption of the autologous lysate, equal numbers of DCs were pulsed with lysate of the tumor cell line AsPc-1 and BxPc-3 for a further five vaccinations. Peripheral mononuclear cells (PMNCs) were harvested after the seventh and compared with PMNCs obtained after the fourteenth vaccination for immunological response. Delayed type hypersensitivity reactivity to DCs pulsed with autologous and allogeneic tumor lysate was also assessed. The patient received a total of fifteen vaccinations. There was no toxicity or evidence of autoimmunity observed. Delayed type hypersensitivity was found to be positive for the autologous as well as the allogeneic tumor lysate pulsed DCs. in vitro cytotoxicity assays demonstrated a dramatic increase of the PMNC killing capacity against the pancreatic cancer cell lines AsPc-1 and BxPc-3 after the fourteenth compared to the seventh vaccination. CT scans revealed a stable disease for six months. The administration of autologous DCs pulsed with allogeneic tumor lysate is non-toxic and suitable for inducing an immunological anti-tumor response. Even though this study was confined to a single patient, the data might open a door for novel immunotherapeutical strategies. PMID- 12579321 TI - Participation of fibroblasts in MMP-2 binding and activation on the surface of oral squamous cell carcinoma cells. AB - The effect of fibroblasts on the activation of matrix metalloproteinase-2 (MMP-2) in oral squamous cell carcinoma (SCC) cells was examined. The plasma membrane of SCC cells failed to bind and activate latent MMP-2. However, treatment of SCC cells with fibroblast-conditioned medium (fibroblast-CM) led to the enhancement of the binding and activation of latent MMP-2 on the cell surface. Moreover, fibroblasts induced the expression of membrane type 1 MMP (MT1-MMP) in SCC cells. MMP-2 activated on the cell surface bound to the surface of SCC cells via alphav integrins. These findings suggest that fibroblasts might facilitate the invasion of SCC cells by increasing the proteolytic activity on the surfaces of SCC cells. PMID- 12579322 TI - Gene expression profile of metastatic colon cancer cells resistant to cisplatin induced apoptosis. AB - The current chemotherapeutic modalities for advanced colorectal cancer are limited. DNA-platinating drugs such as cisplatin have poor efficacy against this malignancy. The aim of this study was to identify genes that render survival advantage after cisplatin treatment in metastatic colon cancer. Cell lines SW480 (primary colon cancer) and SW620 (metastatic lesion from the same patient) were obtained from ATCC. Apoptosis was measured by FACS analysis of cisplatin-treated (0.01-10 micro g/ml) and untreated cells. Simultaneous analysis of approximately 1200 cDNAs was performed by microarray technique on untreated and treated cells from lines. Microarray results were confirmed by RT-PCR. The SW620 cell line was more resistant to apoptosis induced by cisplatin. Western blot analysis revealed equal expression of pro-caspases 3, 8, and 9 in both cell lines. Microarray analysis identified 15 genes and 9 expressed sequence tags (ESTs) significantly altered both by cell type (metastatic vs. non-metastatic) and treatment vs. non treatment. Several of these transcripts are well-characterized genes including MCT, GAD67, P19, GSTM3, Cyclin D1, ATM, and CO-029 that have been implicated in various malignancies. In the present study, we have identified a set of genes responsible for apoptosis resistance following treatment with cisplatin in the late stages of carcinogenesis. Targeting these genes may increase chemotherapy effectiveness in advanced colon cancer and reduce toxicity in normal tissue. PMID- 12579323 TI - The small GTPase RhoA has greater expression in small cell lung carcinoma than in non-small cell lung carcinoma and contributes to their unique morphologies. AB - The two major forms of lung carcinoma, small cell lung carcinoma (SCLC) and non small cell lung carcinoma (NSCLC), are clinically distinct, and are also differentiated by morphology and behavior in culture. SCLC cells have a greater metastatic potential than NSCLC cells in vivo, and exhibit a unique spherical morphology in culture due to their inability to adhere and spread on the substratum. Because the small GTPase RhoA affects metastatic properties and regulates cell morphology, we examined whether differences in RhoA expression and activity contribute to the distinct SCLC and NSCLC phenotypes. We found that the expression and GTPgammaS-dependent activation of RhoA are generally greater in SCLC cell lines (SCC-9, NCI-H69, NCI-H146, and NCI-H345) than in NSCLC cell lines (NCI-H23, NCI-H157, NCI-H520, and NCI-H522). The effects of inhibiting Rho mediated signaling in these cells were investigated by transfecting the cells with cDNA coding for C3 exoenzyme, which ADP-ribosylates and inactivates Rho. Expression of C3 exoenzyme in SCLC cells induces cell-cell compaction, and causes NCI-H345 cells to adhere and spread on collagen IV. In contrast, expression of C3 exoenzyme in NSCLC cells does not induce detectable compaction, but induces cell spreading of NCI-H23 and NCI-H157 cells. Cell proliferation is diminished by Rho inactivation in the majority of the NSCLC cell lines, but not the SCLC cell lines. Expression of p21Cip1/WAF1 is also diminished by Rho inactivation in two of the SCLC cell lines, but is not significantly altered in the NSCLC lines. These results indicate that Rho-mediated signaling may regulate different events in SCLC and NSCLC cells, including adhesion of SCLC cells and proliferation of NSCLC cells. PMID- 12579324 TI - RNA polymerase II stalled on a DNA template during transcription elongation is ubiquitinated and the ubiquitination facilitates displacement of the elongation complex. AB - When mammalian cells are exposed to cisplatin or ultraviolet irradiation, the RNA polymerase II (RNAP II) large subunit becomes ubiquitinated and is subsequently degraded via the proteasomal pathway. Using a DNA template immobilized on magnetic beads in an in vitro transcription reaction, we showed that a pause of the elongating RNAP II complex caused by nucleotide starvation induced the ubiquitination of the stalled RNAP II. The ubiquitinated RNAP II dissociated from the ternary complex when transcription was allowed to resume. The dissociated (free) RNAP II remained ubiquitinated. The proteasome inhibitor MG132 increased the accumulation of ubiquitinated free RNAP II but did not affect the amount of ubiquitinated, template-bound RNAP II, indicating that the ubiquitinated RNAP II was displaced from the template and then degraded by the proteasomes. Our work shows that the elongation complex that was stalled at the template by nucleotide starvation is targeted by the ubiquitin-conjugating system and that ubiquitination facilitates displacement of the stalled RNAP II from the template. Our findings together with the findings by others that DNA damaging agents induced the ubiquitination in mammalian cells that are nucleotide excision repair competent, suggest that the RNAP II ubiquitination may have a role in the regulation of transcription-coupled DNA repair. PMID- 12579325 TI - Chemoimmunotherapy in mice carrying HPV16-associated, MHC class I+ and class I- tumours: Effects of CBM-4A potentiated with IL-2, IL-12, GM-CSF and genetically modified tumour vaccines. AB - The effectiveness of chemoimmunotherapy with ifosfamide derivative CBM-4A and recombinant IL-2, IL-12, GM-CSF, or genetically modified, cytokine-producing tumour vaccines was examined in mice carrying HPV16-associated, MHC class I+ (TC 1), and MHC class I- (MK16) tumours. Intraperitoneal treatment of TC-1 or MK16 tumour-bearing mice with CBM-4A produced a significant tumour-inhibitory effect. When the i.p. treatment of the MHC class I+ TC-1 tumour-bearing mice with CBM-4A was followed by peritumoral s.c. administration of IL-2, IL-12, or both cytokines, the growth of TC1 tumours was inhibited more vigorously than after the chemotherapy alone. In contrast, when the i.p. treatment ofEthe MHC class I- MK16 tumour-bearing mice with CBM-4A was followed by peritumoral s.c. administration of IL-2 or IL-12, the cytokine therapy had no potentiating effect. The only potentiating effect of the MK16 tumour immunotherapy was obtained when the i.p. CBM-4A pretreatment was followed by peritumoral s.c. administration of IL-2 plus IL-12. InEfurther experiments, the TC-1 and MK16 tumour-bearing mice were i.p. pretreated with CBM-4A and then injected s.c., peritumorally, with genetically modified, IL-2 or GM-CSF-producing MK16 tumour vaccines. Whereas both genetically modified tumour vaccines produced a substantial tumour-inhibitory effect in mice carrying TC-1 tumours, no effect of the vaccines was observed in mice carrying MK16 tumour inocula. The systemic effects of local cytokine treatment were examined in mice carrying s.c. MK16 neoplasms, which were pretreated i.p. with CBM-4A, and then injected peritumorally with IL-2 or GM-CSF. Peritumoral administration of GM-CSF had no antimetastatic effect, whereas peritumoral IL-2 administration produced substantial reduction of lung metastases. The systemic antimetastatic effect of IL-2 contrasted with the negligible effect of IL-2 on the s.c. MK16 tumour inoculum. Taken collectively, the results indicate that in mice carrying the MK16 (MHC class I-) tumour, the effects of the adjuvant cytokine therapy were substantially weaker than in mice carrying the TC-1 (MHC class I+) tumour inoculum. PMID- 12579326 TI - Functional form of the effect of the numbers of axillary nodes on survival in early breast cancer. AB - The change in survival in function of the numbers of involved and uninvolved axillary nodes in early breast cancer - i.e. the functional form - was investigated to search for prognostic cutoffs and to assess if ratio-based characterization of node involvement is a significant prognostic factor or not. Women aged 40-69, diagnosed in 1988-1997 with T1-T2 invasive breast carcinoma, who underwent axillary dissection, are selected from the SEER public database. The method determines the functional form by applying smoothed plots to the martingale residuals obtained from a proportional hazards model. The results on 55,267 selected patients find that the ratio of involved nodes on examined nodes, in a multivariate model that takes into account known prognostic factors (age, race, tumor size, topography, histology, grade, hormone receptors), is associated with a relative mortality hazard of 1.012 (95% confidence interval 1.010-1.014; relative increase of mortality of 1.2% for each 1% increase in the percentage of involved nodes). The functional form for the number of uninvolved nodes shows that the relative mortality hazard initially steeply decreases and then tends to level off beyond 5-10 uninvolved nodes. For the number of involved nodes, the relative mortality hazard continues to increase with each involved node without any obvious cutpoint. Even when the number of involved nodes is already large, each additional involved node increases the relative mortality hazard by at least 1.3%. PMID- 12579327 TI - Influence of the intramural innervation on the morphogenesis of the enteroendocrine cells and the genetic construct involved (review). AB - The influence of intramural intestinal innervation on the morphogenesis of the mucosa and in particular of the enteroendocrine cells (EECs) has been studied on male Wistar rats with morphometric, autoradiographic and immuno-histochemical methods in the duodenum, proximal and distal jejunum and ilium before and after myenteric ablation with benzalkonium chloride (BAC). Twenty-one days after denervation alterations were observed in the mucosal structure with thickening of the mucosa, increase in the proliferation rate and with changes in numerical and spatial distribution of D-cells, I-cells, N-cells, glucagon and glicentin i.r. L cells and 5-HT i.r. cells which myenteric ablation caused. Analysis of the genetic constructs involved in the alterations of EECs on the EECs provide evidence for the cAMP responsive elements as the main mediator. PMID- 12579328 TI - Spatial distribution of liposome encapsulated tin etiopurpurin dichloride (SnET2) in the canine prostate: implications for computer simulation of photodynamic therapy. AB - Photodynamic therapy (PDT) is a minimally invasive treatment that can be employed in many human diseases including prostate cancer. PDT for prostate cancer depends on the sequestration of a photosensitizing drug within the glandular tissue. The photosensitizer is subsequently activated by light (usually from a laser) and the active drug destroys tissue. Since prostate cancer is a multifocal disease, PDT must ablate the glandular prostate completely. This will depend on the precise placement of light sources in the prostate and delivery of a therapeutic light dose to the entire gland. Also, sources of light and their spatial distribution must be tailored to each individual patient. The uniform, therapeutic light distribution can be achieved by interstitial light irradiation. In this case, the light is delivered by diffusers placed within the substance of the prostate parallel to the urethra at a distance optimized to deliver adequate levels of light and to create the desired photodynamic effect. To help achieve the uniform light distribution throughout the prostate we have developed a computer program that can determine treatment effects. The program predicts the best set of parameters and the position of light diffusers in space, and displays them in graphical or in numerical form assuming a fixed attenuation coefficient. The two parameters of greatest importance in the computer simulation are attenuation coefficient and critical fluence. Both depend on the concentration of active drug within the prostate gland. It is necessary to know the nature of the spatial distribution of photosensitizer within the prostate to execute computer modeling of PDT with high precision. We found that the concentration of SnET2 is heterogeneous in nature, and is higher in the proximity of the glandular capsule. It is clear therefore that any future attempts of computerized modeling of this procedure must take into consideration the uneven sequestration of photosensitizer and the consequential asymmetrical necrosis of the prostate. PMID- 12579329 TI - Wilson disease protein ATP7B is localized in the late endosomes in a polarized human hepatocyte cell line. AB - Wilson disease is a genetic disorder characterized by the accumulation of copper in the body due to a defect of biliary copper excretion. The gene responsible for Wilson disease has been cloned, however, the precise localization of this gene product ATP7B, a copper-transporting ATPase, is still controversial. We examined the localization of ATP7B by expressing a chimeric protein, ATP7B-tagged with green fluorescent protein (GFP) (GFP-ATP7B), in HEK293, Hep3B and a highly polarized human hepatocyte line (OUMS29). Intracellular organelles were visualized by immunofluorescence microscopy. The effects of bathocuproine disulfonate, a copper chelator, and copper sulfate were examined. GFP-ATP7B colocalized with a late endosome marker, but not with endoplasmic reticulum, Golgi, or lysosome markers in a copper-depleting condition. Treatment with copper sulfate did not affect the localization of ATP7B. ATP7B is localized in the late endosomes in both copper-depleting and copper-loaded conditions. ATP7B seems to translocate copper from the cytosol into the late endosomes, and copper may be excreted to bile via lysosomes. We believe that the disturbed incorporation of copper into the late endosomes caused by mutated ATP7B is the main defect in Wilson disease. PMID- 12579331 TI - Epidermal growth factor receptor expression in human pancreatic cancer: Significance for liver metastasis. AB - Pancreatic cancer is a malignant tumor with an extremely poor prognosis. The mechanisms of the aggressive growth and metastasis are not yet extensively understood. Over-expression of epidermal growth factor receptor (EGFR) was suggested to be associated with malignant transformation of pancreatic cancer. We examined EGFR expression in 77 cases of invasive ductal adenocarcinoma of the pancreas, and analyzed the relation between the EGFR expression pattern and clinicopathological factors. EGFR immunoreactivity was detected in 41.6% (32/77) of human pancreatic cancers; i.e. diffuse expression in 32.5% (25/77) and focal expression in 9.1% (7/77). The EGFR expression was associated with gender (p<0.05), histological differentiation (p<0.05) and metastatic status of TNM classification (p<0.01). The observations suggested that EGFR expression plays important roles in metastasis, especially liver metastasis and recurrence of human pancreatic cancer. PMID- 12579330 TI - Role of apoptosis inhibition in various chondrocyte culture systems. AB - Apoptosis may limit the utility of cell-based therapies for articular cartilage disorders. We tested the hypothesis that chondrocyte apoptosis can be reduced by optimizing the conditions employed to expand chondrocyte numbers in culture. Chondrocyte apoptosis was examined in monolayer and suspension culture, in the presence of fetal bovine serum (FBS) or autologous serum, and for culture periods of 2 or 4 days. The effect of these variables was assessed by measuring cell viability, Annexin V labeling and mitochondrial membrane potential. After 2 days of culture, the greatest increase in viable cell number (3.7-fold) occurred in monolayer cultures with autologous serum. After 4 days of culture, the greatest increase in cell number (9.0-fold) occurred in monolayer cultures supplemented with FBS. By Annexin V staining, the proportion of cells undergoing apoptosis after 2 days was not affected by the type of serum used or by culture in monolayer versus suspension. After 4 days of culture, the proportion of apoptotic cells was significantly reduced (35% to 13%, p<0.02) in suspension cultures with autologous serum. Apoptosis assessed by loss of mitochondrial membrane potential was decreased in the presence of autologous serum. These data suggest that suspension culture with autologous serum is useful in simultaneously maintaining cell proliferation and minimizing apoptosis in cultured human articular chondrocytes. PMID- 12579332 TI - Successive increases in human cyclin A1 promoter activity during spermatogenesis in transgenic mice. AB - The human cyclin A1 gene is highly expressed in pachytene spermatocytes and is essential for spermatogenesis. To analyze mechanisms of cyclin A1 gene expression in vivo, we cloned a 1.3 kb fragment of the promoter upstream of the cDNA of enhanced green fluorescent protein (EGFP). Four lines of transgenic mice were generated that carried the transgene. Cyclin A1 promoter activity in the organs of the transgenic mice was analyzed using fluorescence microscopy and flow cytometry. Expression of EGFP was seen in male germ cells of all four murine lines. Spermatogonia at the basal membrane expressed low levels of EGFP, but bright green fluorescence was present in spermatocytes entering meiosis. Interestingly, a further sharp increase in EGFP expression was found in spermatocytes approximately at the stage of the first meiotic division. EGFP levels stayed high thereafter and EGFP was present in mature spermatozoa. A portion of c-kit expressing cells in the testis also expressed EGFP indicating cyclin A1 promoter activity in a subpopulation of spermatogonia. These data suggest that cyclin A1 is active not only in pachytene spermatocytes but also in earlier phases of spermatogenesis. PMID- 12579334 TI - Genetic susceptibility for emphysematous changes of the lung in Japanese. AB - Risk of chronic pulmonary emphysema from exposure to tobacco smoke varies widely from person to person, depending in part on the status of particular genes and acquired susceptibilities. Certain genes determine how cells activate and/or detoxify tobacco smoke products. We aimed to determine whether any genetic susceptibility exists in the development of emphysematous changes confirmed by chest computed tomography (CT). Genotypes of various enzymes involved in the activation or detoxification of tobacco smoke, epoxide hydrolase (EPHX1), cytochrome P450s (CYP1A1 and CYP2E1), glutathione S-transferases (GSTM1/P1/T1), and hemoxygenase-1 (HMOX1), were determined by PCR-based assays in a total of 235 heavy smokers (Brinkman index >/=400). They were divided into two groups according to the CT findings: 63 and 172 subjects with and without emphysematous changes, respectively. Although the allele frequency of any genetic polymorphism was not statistically different between the two groups, the frequency of the individuals having combination of the genotype representing very slow activity for epoxide hydrolase and at least one allele with large size of (GT)n repeats in the HMOX1 gene promoter region was higher in the subjects with emphysematous changes (p=0.03; odds ratio 2.8; 95% CI = 1.07-7.5) among the stratified individuals (age >/=51 years). These findings suggest that combination of several polymorphisms in the enzymes that activate or detoxify the tobacco smoke, such as EPHX1 and HMOX1, might be associated with its affects on the development of emphysematous changes of the lung. PMID- 12579333 TI - Effect of red wine and wine polyphenol resveratrol on endothelial function in hypercholesterolemic rabbits. AB - The effect of red wine and wine polyphenol resveratrol on endothelial function was investigated in experimental hypercholesterolemic rabbits. Endothelial function as measured by flow-mediated dilation (FMD) in the femoral artery was 19.28+/-2.81% in control animals fed a regular diet. In contrast, rabbits fed a high-cholesterol (1.5%) diet showed a reduced endothelial function, as revealed by a 25% reduction in the measured FMD. Intragastric feeding of resveratrol (3 mg/kg/day), red wine (4 ml/kg/day), dealcoholized red wine (4 ml/kg/day), for 12 weeks in hypercholesterolemic rabbits significantly mitigated the reduction in endothelial function, and resulted in FMD values of 14.52+/-0.60, 18.95+/-2.30, 17.58+/-1.43, and 18.80+/-3.94%, respectively. Measurement of plasma endothelin 1 (ET-1) and nitric oxide (NO) levels showed that feeding a high-cholesterol diet significantly increased plasma ET-1 levels (from 51.4+/-17.6 to 96.9+/-24.3 pg/ml), and decreased plasma NO concentration (from 104.6+/-18.5 to 67.7+/-16.1 pg/ml). With administration of resveratrol, red wine, or dealcoholized red wine, plasma ET-1 levels statistically decreased, in parallel with a significant elevation in NO levels. These results provide in vivo evidence suggesting that resveratrol and red wine improve endothelial function, which may be one of the mechanisms by which this red wine polyphenol exerts its alcohol-independent cardioprotective effects. PMID- 12579335 TI - T cell epitopes of type II collagen in HLA-DRB1*0101 or DRB1*0405-positive Japanese patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a T cell-mediated autoimmune disease, but target antigens (autoantigens) responsible for T cell activation remain unclear. Type II collagen (CII) is a candidate autoantigen that is largely confined to the articular cartilage. To investigate whether CII is an important antigen in patients with RA, we examined peripheral blood T cell reactivity to CII in HLA DRB1*0101 and DRB1*0405-positive RA patients. Reactivities to candidate T cell epitopes of CII were also examined. Peripheral blood T cell reactivity to CII and CII peptides (256-271, 429-442, 593-610, 1064-1081) were detected by measurement of IL-2, IFN-gamma, and IL-4 in culture supernatant of PBMC after in vitro antigen stimulation. Cytokine concentration was measured by ELISA. In DRB1*0101 positive patients, T cell reactivity to CII as detected by measurement of IL-2 production in culture supernatant, was present in 4 out of 9 patients. IL-2 production upon stimulation with CII 256-271 peptide was found in all of these 4 patients. In DRB1*0405-positive patients, high frequency of positive T cell response to CII was detected in 9 out of 11 patients. IFN-gamma production was also detected in 4 out of 6 patients producing IL-2 by stimulation with CII. T cell response to CII 256-271 and/or CII 1064-1081 was detected in these patients. In DRB1*0101-positive RA patients, CII 256-271 peptide might function as a T cell epitope, whereas either CII 256-271 or CII 1064-1081 peptide may be a major T cell epitope in DRB1*0405-positive RA patients. In DRB1*0405-positive RA patients, CII reactive T cells might play a crucial role in the development of RA through IFN-gamma production. PMID- 12579336 TI - Expression of copper-transporting P-type adenosine triphosphatase in human esophageal carcinoma. AB - A major obstacle in the treatment of esophageal carcinoma is the intrinsic/acquired resistance to cisplatin-based chemotherapy. Copper transporting P-type adenosine triphosphatase (ATP7B) has been reported to be associated with cisplatin resistance in vitro. However, the clinical significance of this transporter has not previously been addressed. Our goal was to investigate if ATP7B is expressed in esophageal carcinoma and whether its expression correlates with reduced responsiveness to cisplatin treatment. We retrospectively examined the expression of ATP7B in primary esophageal carcinoma and its association with chemotherapeutic effect. Tissues were surgically removed from 17 esophageal carcinoma patients. Twelve of them received cisplatin-based chemotherapy before surgery. We performed immunohistochemical analysis of ATP7B using a monoclonal antibody against ATP7B in 17 esophageal carcinomas. A variable degree of cytoplasmic staining of tumor cells was observed in 76.5% (13/17 cases) of the analyzed carcinomas. ATP7B expression was not observed in adjacent non neoplastic tissues. ATP7B positivity was not significant in gender, age, histopathological grading or TNM categories. Patients with ATP7B-positive tumors tended to have an inferior response to chemotherapy compared with the patients with ATP7B-negative tumors. These findings suggest that overexpression of ATP7B in esophageal carcinoma could be associated with unfavorable clinical outcome in patients treated with cisplatin-based chemotherapy. Therefore, ATP7B gene expression might be considered as a chemoresistance marker for cisplatin in the patients of esophageal carcinoma and provider of important information on the strategy against esophageal carcinoma. PMID- 12579337 TI - Telomerase activity, expression of Bcl-2 and cell cycle regulation in doxorubicin resistant gastric carcinoma cell lines. AB - As telomeres play a role in protecting DNA, there is the possibility that telomerase activity is involved with cellular response to DNA-damaging agents. This study was designed to investigate the association between telomerase and the doxorubicin altered cell cycle in drug resistant gastric carcinoma cell lines. Three doxorubicin resistant gastric carcinoma cell lines and their parent cell lines (SNU-1, SNU-16 and SNU-620) were incubated with doxorubicin at the final concentration induced resistance and ten times final concentration for 24 h. Telomerase activity and hTERT mRNA expression were lowered by doxorubicin treatment in parent cell lines, but in drug resistant cell lines, telomerase activity and hTERT mRNA expression were not repressed by doxorubicin treatment. Bcl-2 protein expression, which is known to regulate telomerase activity, did not change in doxorubicin resistant cell lines but decreased in parent cell lines by doxorubicin treatment. Cell cycle analysis revealed that the parent cell lines had an increased fraction of cells in G2/M phase after doxorubicin treatment and doxorubicin resistant cell lines had maintained fractions in G0/G1 phase. Doxorubicin treatment did not alter cyclin B or cdc2 protein level, which is known as the essential component of G2/M transition. G2/M arrest in the parent cell lines was associated with an increase in inhibitory phosphorylation of Tyr15 on cdc2. In summary, the parent cell lines showed G2/M arrest and a reduction of telomerase activity after doxorubicin treatment. In contrast, reduced telomerase activity, Bcl-2 expression and G2/M arrest after doxorubicin treatment did not appear in resistant cell lines. Therefore, relative resistance to doxorubicin may be related to high levels of bcl-2 or intact cell cycle and consequently high telomerase activity. PMID- 12579339 TI - Leukemia inhibitory factor regulates cell survival of normal human endometrial stromal cells. AB - Leukemia inhibitory factor (LIF) is produced locally in decidual tissues, but its direct effects on endometrial stromal cells have not been well characterized. In this study, we examined the direct effects of LIF on normal human endometrial stromal cells using an in vitro decidualization assay system with 8-Br-cAMP, a decidualization inducer. We found no effects of LIF on cell viability and prolactin secretion of unstimulated endometrial stromal cells. LIF dose dependently enhanced cell viability, but not prolactin secretion of 8-Br-cAMP stimulated cells. LIF dose-dependently enhanced the viability of stromal cells co stimulated with 8-Br-cAMP and LIF without any significant effect on PRL secretion from the cells. Further, the extracellular matrix did not affect these stimulatory effects of LIF on cell viability of 8-Br-cAMP-stimulated endometrial stromal cells. These results indicate that LIF enhances the cell viability of PRL non-secreting 8-Br-cAMP-stimulated stromal cells, and that the cell survival signals generated by LIF are independent of those generated by the extracellular matrix. LIF produced locally in decidual tissues may enhance cell viability in certain activated endometrial stromal cells in an autocrine or paracrine manner, and possibly protect against cell damage during embryo implantation and trophoblastic invasion. PMID- 12579338 TI - Mucosal permeability regulates receptor binding of luminal epidermal growth factor in the adult rat intestine. AB - Epidermal growth factor (EGF) stimulates repair in the damaged intestine, but its role in the normal intestine is not clear. Because EGF receptors are found on the basolateral surface but not the luminal surface, we hypothesized that mucosal permeability regulates EGF binding. Adult male rats were divided into 3 groups, one that was fed normal chow (the control), one that was starved for 4 days, and one that was given methotrexate (MTX) intragastrically (10 mg/kg/day for 3 days). The rats were sacrificed and everted sacks of the jejunum were made and incubated in EGF solution. Western blot analysis of mucosal homogenates showed that the amount of phosphotyrosyl EGF receptor in the starved and MTX-treated groups was, respectively, about 1.5 times and 2 times that in the control group. The mucosal permeability in the starved and MTX treated groups also increased and varied directly with the amount of phosphotyrosyl EGF receptor. These results suggest that in the adult rat intestine, luminal EGF may play a role only under tissue damage, where enhanced permeability permits the EGF to pass through the mucosa and bind to its receptor on the basolateral membrane. PMID- 12579340 TI - Gender dimorphism in neutrophil priming and activation following trauma hemorrhagic shock. AB - Although gender influences T-cell, macrophage and organ functions following trauma-hemorrhage and resuscitation (T-H), it remains unknown whether it also influences polymorphonuclear cell (PMN) activity under such conditions. To study this, proestrus female and male Sprague-Dawley rats underwent trauma-hemorrhage followed by fluid resuscitation. Circulating PMNs were assessed for superoxide (O2-) and elastase production and tissues were analyzed for myeloperoxidase (MPO) activity and TBARS (thiobarbituric acid reactive substances) as a marker of oxidative injury, at 2 and 24 h after resuscitation. PMA stimulated O2- production was not influenced by T-H or gender. In contrast, fMLP-stimulated O2- and LPS-stimulated elastase release by PMNs from male T-H rats was greater than that of females. A significant MPO activity and TBARS in tissues of both male and female rats was induced; however, MPO activity and TBARS levels were higher in males following T-H. Levels of the chemokine CINC-1 were elevated in the lungs of male, but not of proestrus females after T-H. Thus, decreased PMN priming and activation in proestrus females, compared to males, occurs following T-H resulting in decreased cellular injury and organ damage that is likely to contribute to improved outcome under those conditions. PMID- 12579341 TI - Percutaneous transluminal angioplasty against arteriosclerosis obliterans in dialysis patients. AB - The incidence of peripheral arteriosclerosis is on the increase in chronic hemodialysis patients. Recently, the intervention (IV) treatment is conducted to deal with this problem. IV was performed in 4 dialysis patients against the complication of arteriosclerosis obliterans (ASO) but the result was unsuccessful in 3 of them. These 3 failure cases were investigated to find the problems associated with percutaneous transluminal angioplasty (PTA). Cases 1, 2 and 3 had intermittent claudication while case 4 had gangrenous toes as the major complaint. The symptoms in these cases were attributable to 90-100% stenosis and occlusion of superficial femoral artery, bilateral iliac arteries, bilateral superficial femoral-popliteal artery, branch of right iliac artery and left iliac artery region, respectively. IV was successful in case 1 but failed in cases 2 and 4 because the catheter itself did not go through due to the severe stenosis of vessel or the procedure of forcefully dilating the vessel caused dispersion of minute thrombi. In case 3, acute myocardial infarction occurred at 10 h after successful IV, resulting in sudden death. In view of the extent of invasion, IV is a treatment method selected against ASO in dialysis patients. However, the method has a high risk of causing thrombus formation, vessel rupture and organ failure. In this regard, it is advisable to evaluate the systemic condition and conduct IV if the extent of stenosis is mild. PMID- 12579343 TI - Effect of cryopreservation on in vitro clonal growth of cordonal blood cells. AB - Cordonal blood (CB) is today recognized as a potentially important source of hematopoietic stem cells (SCs) for allogeneic transplantation, and to this task it would be of extreme importance to have the possibility of using cryopreserved CB units. Hence we investigated whether freezing and thawing alter the viability of CB hematopoietic cells. Mononuclear cells, recovered from fresh CB units by density-gradient centrifugation, were partly frozen and then thawed and partly immediately utilized for clonogenic tests. The cells were cultured in H4330 (C group) or H4434 (H4330 added with SC clonogenic growth factors) (C+ group) semisolid media added or not with serum-free Dulbecco modified Eagle medium (DMEM/sf). Cells seeded on H4330 were exposed to the conditioned supernatants from two human-embryo liver cell lines, which have been previously found to stimulate clonal growth of fresh CB hematopoietic cells. As expected, the number of colony-forming units (CFU) was higher in C+ than C group, and was not influenced by the addition of DMEM/sf. CFU number was higher in cryopreserved than fresh cells in both C and C+ culture groups. Conditioned supernatants from both cell lines stimulated clonal growth in both fresh and cryopreserved cell cultures. These findings indicate that cryopreservation and thawing do not alter the viability of CB SCs, but, on the contrary, improve their basal and cytokine stimulated clonal growth, probably by negatively selecting SCs among the mononuclear CB cell population. PMID- 12579342 TI - Bax cleavage implicates caspase-dependent H2O2-induced apoptosis of hepatocytes. AB - Oxidative stress plays an important role in the development of ischemia/reperfusion (I/R)-induced apoptosis of hepatocytes. We aimed to examine the involvement of caspases and calpains in H2O2-induced hepatic cell apoptosis. TUNEL-positive apoptotic cells appeared in parallel with poly(ADP-ribose) polymerase (PARP) cleavage and procaspase-3 proteolysis by H2O2 treatment in a dose-dependent manner (250-1,000 micro M). Bcl-xL and intact Bax expression levels decreased when H2O2 was >250 micro M. The cleaved form of Bax appeared prior to caspase-3 activation, increasing in a dose-dependent manner. A pan caspase inhibitor, Z-VAD-fmk, completely blocked H2O2-induced procaspase-3 proteolysis and PARP cleavage without changing Bax cleavage, but partially attenuated H2O2-induced apoptosis. Calpeptin, a calpain inhibitor, did not inhibit caspase-3 activation, Bax cleavage or apoptosis. Our results indicate that Bax cleavage is upstream signal of caspase-dependent apoptosis in hepatocytes exposed to H2O2, but not independent upon calpain. Molecular targeting of Bax cleavage may allow the development of strategies to prevent hepatic I/R injury. PMID- 12579344 TI - SNPs and haplotypes: genetic markers for disease and drug response (review). AB - The genetic make-up of an individual not only determines disease susceptibility but also response to drug treatment. It is because of this reason tremendous progress has recently been made in cataloging human sequence variations with an intention such that a high-density map will provide necessary tools to develop genetic-based diagnostic and therapeutic options. The availability of the human DNA sequence, its variation between individuals and the functional understanding of genetic determinants between individuals may enable pharmaceutical companies to discover safer and effective drugs. This will provide physicians with better tools to select the most appropriate treatment for each patient and avoid drugs that cause adverse effects. When a personalized medication becomes a more realistic and beneficial treatment, every newborn child in the neonatal unit will probably be genotyped as a routine procedure for improved treatment. This newly developed toxicogenomic or pharmacogenomic field is rapidly advancing and may change the practice of medicine in the future, providing personalized medicine for each individual. PMID- 12579345 TI - Identification and characterization of human SNAIL3 (SNAI3) gene in silico. AB - Human SNAIL1 (SNAI1) protein encoded by SNAI1/SNA gene represses transcription of E-cadherin/CDH1 gene. Human SNAIL2 (SNAI2) protein encoded by SNAI2/SLUG gene induces the first phase of epithelial-mesenchymal transition (EMT), including desmosome dissociation, cell spreading, and initiation of cell separation. Here, we have identified human SNAIL3 (SNAI3) gene using bioinformatics. Human SNAI3 gene, consisting of at least three exons, spans around the nucleotide position 320214-328221 of human reference genomic contig NT_010404.8 in the reverse orientation. SNAI3 gene, was located between KIAA0233 gene and CBFA2T3 gene in human chromosome 16q24.3, a region affected in breast cancer, gastric cancer, hepatocellular carcinoma, ovarian cancer, and therapy-related myeloid leukemia with t(16;21)(q24;q22) translocation. Human SNAI3 gene was found to encode 292 amino-acid polypeptide with the N-terminal SNAG domain and five zinc finger domains. N-terminal SNAG domain was identified in zinc finger proteins SNAI1, SNAI2, SNAI3, SCRATCH (SCRT1), GFI1, and GFI1B. ATP/GTP binding site was identified in SCRT1, GFI1 and GFI1B, but not in SNAI1, SNAI2 and SNAI3. Phylogenetic analysis of human zinc finger proteins with SNAG domain revealed that SNAI1, SNAI2 and SNAI3 were more closely related. These results clearly indicate that SNAI1, SNAI2 and SNAI3 constitute a subfamily among SNAG zinc finger proteins. Human SNAI3 mRNA was expressed in skin melanotic melanoma, lung epidermoid carcinoma, and germ cell tumor. Because SNAG zinc-finger proteins are transcriptional repressors implicated in carcinogenesis and embryogenesis, SNAI3 gene might be a potent target of pharmacogenomics in the field of oncology and regenerative medicine. PMID- 12579346 TI - Apoptosis of bladder cancer cells induced by short-term and low-dose Mitomycin-C: potential molecular mechanism and clinical implication. AB - Adjuvant chemotherapies have been used to prevent recurrence of bladder transitional cell carcinoma (TCC), but their efficacies are variable due to the side effects of anti-cancer agents and the drug resistant property of the target cells. To provide experimental evidence for improving clinical management of TCCs, two human TCC cell lines, EJ and BIU, were treated for 1 or 2 h by 50, 100, 150 and 200 micro g/ml, respectively and their growth and death patterns were elucidated in 12-h intervals. The status of Fas, FasL and caspase-3 in the two cell lines were analyzed with immunocytochemical staining and Western blot hybridization, and their potential link to MMC-induced cell death was investigated by treating the cells with anti-Fas antibody (150 ng/ml) and by incubating the cells with an inhibitor of caspase-3 related proteases Ac-DEVO-CHO (250 micro M) 1 h before 100 micro g/ml MMC treatment. The results demonstrated that the lower dose (100 micro g/ml) and short-term (1 h) MMC treatment could induce sufficient apoptosis in EJ and BIU populations within 48 or 60 h. Constitutive soluble and membrane Fas and FasL were found in both cell lines, and caspase-3 could be upregulated after MMC treatment. Anti-Fas antibody could commit the target cells to die of apoptosis, while Ac-DEVD-CHO inhibited MMC induced apoptosis. Our data thus suggest that MMC-induced apoptosis in EJ and BIU cells is mediated by Fas and upregulation and activation of caspase-3 is an essential element for the apoptotic process. Reduced dose and short-term MMC strategy would be of practical value either in determining the apoptotic susceptibility of individual TCC cases or in the clinical instillation of urothelial cancers. PMID- 12579347 TI - Stem cell transplantation for treatment of liver diseases: from biological foundations to clinical experience (review). AB - The insufficiency of liver functions remains one of the major causes of death in different regions of the world. This syndrome is an ultimate outcome of irreversible damage of liver parenchyma by a variety of stimuli such as environmental toxins and microbial metabolites. The acute and/or chronic inflammation caused by these agents leads to massive apoptosis of hepatocytes, highly specialized polyfunctional epithelial cells. A decrease in the number of hepatocytes below the critical level dictates the necessity of supplanting the exogenous viable cells. Such cells must fulfill two major requirements: i) they should be capable of proliferating in the recipient's liver, and ii) they should differentiate into mature (i.e, functionally competent) hepatocytes. Evidence is growing in support of the role of early pluripotent precursors (stem cells of various origin) as a unique source for transplantation. This review analyses the state-of-the-art in the fields of developmental biology, cytology, molecular physiology and biochemistry of fetal stem cells with the emphasis on their use in experimental models of liver insufficiency. These biological considerations validate the therapeutic manipulations with fetal cells in clinical trials. PMID- 12579348 TI - The effects of interferon-gamma and transforming growth factor-beta on adherence and survival of group B Streptococcus type III strains in ECV304 cells. AB - Group B streptococci (GBS) are an important cause of neonatal sepsis, pneumonia and meningitis. In some newborns, GBS sepsis may have a severe course, including septic shock with high mortality rate, whereas other newborns are colonized with GBS on their surfaces without any clinical signs of bacterial infections. Interferon (IFN)-gamma is produced in neonatal GBS sepsis, and transforming growth factor (TGF)-beta is also found in the uterus. The involvement of IFN gamma and TGF-beta in the earliest phase of infection might be a determinant of susceptibility and/or progression of infection in vivo. The aim of this study was to assess the effect of IFN-gamma and TGF-beta on adherence and intracellular viability in ECV304 cells of GBS serotype III isolated from cerebrospinal fluid (CSF) and vagina (strains 90356 and 80340, respectively). Interaction of GBS ECV304 cells showed that the CSF isolate exhibited a more efficient adherence mechanism than the vagina isolate (P<0.001). Intracellular viability was observed for the CSF 90356 isolate within 2 h incubation. Results suggest the expression of additional bacterial virulence factors that favor some GBS type III strains to cause invasive disease. Detection of genotypic virulence marker (162-kb) in the CSF 90356 isolate by PFGE emphasizes the high risk of invasive infection by some GBS-III strains. Treatment of ECV304 cells with IFN-gamma and/or TGF-beta increased adherence of both GBS strains (P<0.001). Intracellular survival of the CSF 90356 isolate was observed after 24 h incubation following treatment of ECV304 cells with IFN-gamma and TGF-beta. Our data suggest that both IFN-gamma and TGF-beta may favor virulence of GBS strains. Variation of IFN-gamma and TGF beta producing capacity of host cells of different individuals may influence development of invasive disease by GBS-III. PMID- 12579350 TI - Humane endpoints in the efficacy testing of swine erysipelas vaccines. AB - For licensing the efficacy of vaccines for veterinary use has to be demonstrated by well-controlled laboratory experiments in which vaccinated and untreated animals of the target species are challenged. Erysipelas challenge tests cause extreme suffering of the unprotected animals with high fever, apathy, large skin lesions, and even death. This paper describes a standardised procedure for the vaccination challenge test and gives due consideration to the welfare of the animals. By monitoring and using clinical signs observed during the test it is possible to minimise animal pain and distress, thus preventing unnecessary animal suffering. PMID- 12579349 TI - Albumin effects on drug absorption and metabolism in reconstructed epidermis and excised pig skin. AB - To replace animal experiments for the risk assessment in cutaneous absorption European and Non-European regulatory authorities urge to develop in vitro test systems. A variety of methods have been established which, however, vary in several respects, such as the nature of the barrier and used media. Since both affect drug uptake we have compared the influence of albumin (BSA) in the acceptor medium on the permeation and metabolism of highly lipophilic steroidal drugs, testosterone and prednicarbate (PC). Surprisingly, the addition of BSA to the acceptor medium slightly reduced the steroid permeation, especially when formulations of poor PC uptake were tested. Moreover, with slow drug permeation the metabolite pattern changed as compared to PC metabolites to be found in albumin-free acceptor medium. This was clearly less the case with PC incorporated into newly developed solid lipid nanoparticles accelerating PC uptake about fourfold. The penetration of testosterone was not influenced by BSA in the acceptor medium. Summarising, these results contribute essentially to the development of appropriate in vitro methods for testing of the cutaneous absorption of drugs, ingredients of cosmetics and for the risk assessment of xenobiotics, pesticides and biozides. PMID- 12579351 TI - [Totally artificial training model for coronary heart surgery: the renunciation of animal experiments?]. AB - AIM: Animal protection laws will lead to stricter and more selective criteria thus resulting in a decline of available animals. Yet to train cardiac surgical skills a totally artificial training model was developed. DESCRIPTION OF THE TRAINING MODEL: The model is based on differently hardened polyurethane. Cover is a 1:1 replica of the human thoracic wall. Disposable coronaries are integrated in the heart-model. Vessels and part of the ascending aorta can be rinsed. By means of a newly designed air-pump stroke volume, heart-rate and rhythm can be adjusted. EXPERIENCES: Set-up of the model is easy and quick. Accustomed instruments can be used. Handling of artificial tissue is nature-like. Degree of difficulty is dependent on stroke volume, heart rate, arrhythmia, vessel-size and vessel-quality. CONCLUSION: The phantom helps to achieve confidence in coronary revascularisation. It facilitates an accompanying training for the less-trained as well as the skilled surgeon. The nature-like characteristics will help to reduce animal experiments in future. PMID- 12579352 TI - Extraction of a monospecific Coombs-reagent from chicken eggs. AB - During the last ten years the extraction of specific antibodies (ab) from the yolk of eggs of immunised chickens is more and more accepted as an useful alternative to the immunisation of mammals. The subject of this work is the immunisation of chickens with human IgG and the extraction of specific anti human IgG ab from egg yolk in order to obtain monospecific Coombs reagent. 12 Leghorn hens (25 weeks old) were immunised with intact human IgG (INTACGLOBIN). The chickens were immunised with 100 microg IgG/animal once per week for a period of seven weeks. The highest titre was observed after the 5th immunisation, the following immunisations achieved no further titre increase. The IgY purification was performed according to the method of Akita and Nakai (1993). The resulting IgY preparation was tested for the presence of hetero-agglutinine by means of direct agglutination using human erythrocytes of all blood groups. Thereafter 58 blood donors were tested by means of direct or indirect Coombs-test using a reference reagent (DAKO) and a Coombs reagent isolated from chicken eggs (IgY antibodies). No differences have been found between the results obtained using both Coombs reagents. Presented results show that there is a possibility to produce Coombs-reagent in chickens. Advantages of this method are: 1) non invasive antibody sampling by egg collection instead of bleeding the animal (refinement of antibody production); 2) decreasing amount of animals necessary to produce high amounts of reagent; 3) IgY-preparation contains no hetero agglutinine in contrast to serum ab from mammals, therefore additional step in reagent production e.g. the absorption of hetero-agglutinins is not necessary. PMID- 12579353 TI - [Animal experimentation statistics in Germany: 2 million mark exceeded]. PMID- 12579354 TI - [The 'sewage unit fish' must further die]. PMID- 12579355 TI - Oxygen stress and adaptation of a semi-aquatic plant: rice ( Oryza sativa). AB - One of the major abiotic stresses that affects plant growth and development is anoxia or hypoxia. Rice is a semi-aquatic plant bestowed with the capability of overcoming oxygen limitation for a considerable period of time. For instance, it can withstand submergence stress either by inherent metabolic adaptations (resistant type), or by keeping its leaves above the water surface by continuously elongating the stem (avoiding type). In the former case, an interplay of several metabolic pathways engaged in anaerobic fermentation keeps the submerged plant alive for a certain period of time. In the latter type, also known as deepwater rice, continuous stem elongation brought about by a series of reactions in planta enables the shoot to remain above the water surface and thus maintain respiration and photosynthesis. However, the earliest event, i.e., sensing the oxygen level that brings about all the changes, has not been clearly understood. This paper intends to evaluate the metabolic adaptations of rice plants to oxygen constraints. PMID- 12579357 TI - Leaf anatomy of a rheophyte, Dendranthema yoshinaganthum (Asteraceae), and of hybrids between D. yoshinaganthumand a closely related non-rheophyte, D. indicum. AB - The rheophyte Dendranthema yoshinaganthum(Makino ex Kitam.) Kitam. is endemic to a region along the Naka River in Shikoku, Japan. Resembling other rheophytes that have adapted to environments along rivers, D. yoshinaganthumhas narrow leaves. In the native habitat of D. yoshinaganthumin Tokushima, Japan, natural hybridization with Dendranthema indicum (L.) Des Moul. has resulted in a species complex with conspicuous variation in leaf shape and, in particular, in leaf width. In this study, D. yoshinaganthum and hybrids with natural variation in leaf shape were cultivated under identical conditions and the anatomy of their leaves was examined. Variation in leaf width in the species complex was attributable, for the most part, to variation in the number of cells per leaf lamina and also, to some extent, to variation in cell size. This is the first report of natural variation in leaf width that is attributable to variation in both the size and the number of leaf cells. PMID- 12579356 TI - Hybridization between European and Asian dandelions ( Taraxacum section Ruderalia and section Mongolica)2. Natural hybrids in Japan detected by chloroplast DNA marker. AB - Natural hybridization in Taraxacum between native sexual diploids and introduced agamospermous triploids occurring in Japan was studied by means of chloroplast DNA (cpDNA) marker. We first determined the nucleotide sequences between trnT (UGU) and trnF (GAA) of cpDNA for 22 plants obtained from Japan and Europe. The sequences analyzed were about 1,574 base pairs long. Among all accessions, the total numbers of polymorphic characters were 56 nucleotide substitutions, three insertions/deletions (ins/dels), and one repeat number polymorphism of mononucleotide motif. Of these polymorphic characters, four nucleotides and one ins/del were applicable in the discrimination between Japanese and European taxa of dandelions. We selected the ins/del in an intergenic region between trnL (UAA) 3' exon and trnF (GAA) as a cpDNA marker. Using a newly developed cpDNA marker, 225 plants of putative Taraxacum officinale collected from 11 populations in Niigata City were investigated. Eighty-two percent of them showed a Japanese haplotype of cpDNA, and they were regarded as hybrids. Compared with the previous studies, it is likely that the prevalence of the hybrid plants is a general phenomena at least in urban areas in Japan. The validity of the cpDNA marker for screening Taraxacum hybrids is discussed. PMID- 12579358 TI - Pollen-pistil relationships and pollen size-number trade-off in species of the tribe Lycieae (Solanaceae). AB - Pollen volume may be involved in different associations with other floral traits. Particularly, the literature indicates that pollen volume can be implicated in a functional relationship with pistil length, and that it may be affected by a trade-off with pollen number because of the subdivision of limited resources. To assess these associations, pollen volume was subjected to correlation analyses with pollen number and pistil length in 20 Argentinean and Chilean taxa of the monophyletic tribe Lycieae. Depending on the mechanisms operating on pollen size and number, the variability of these traits may be different. Therefore, their coefficients of variation were compared. Pistil length and pollen volume showed a strong positive correlation. In contrast, pollen grain size and number were significantly correlated at neither inter- nor intraspecific levels. Results suggest that pollen size and pistil length may co-evolve. The central role of this interaction is discussed. The lack of a correlated variation in pollen size and number may be related to the similar constraints experienced by the species and/or because constraints are operating at the plant level and not at the species level. Lower variance in pollen size compared with pollen number denotes that pollen size may be the main trait subjected to natural selection. PMID- 12579359 TI - Demography and matrix analysis on a natural Pterocarya rhoifolia population developed along a mountain stream. AB - The population structures and dynamics of a dominant riparian canopy species, Pterocarya rhoifolia (Juglandaceae), were analyzed based on the census data collected for the 7 years from 1989 through 1996. A study plot was established over an environmental gradient that included the lower hill-slope, river terrace, and low-level floodplain of a riparian area within a 2.8-ha cool-temperate forest stand. Spatial analyses of the demographic data showed that there were significant differences in recruitment, growth, and death processes among the three habitats. The slope and terrace habitats were the severest for the pre reproductive stage and recruitment process, respectively. A topographically combined projection matrix was constructed for the life-history processes of the three subpopulations, which clearly revealed that the floodplain and terrace subpopulations constituted the mainland source populations, whereas the slope subpopulation was an island sink population. The whole Pterocarya population linked by seed flow showed an increase in population size (lambda=1.052). The elasticity analysis showed that the sum of the elasticity values was zero, 0.14, and 0.85 in the slope, terrace and floodplain subpopulations, respectively. This fact clearly indicates that the role of the floodplain subpopulation as a source population is six times as large as that of the terrace subpopulation. PMID- 12579360 TI - Bumblebee-pollination and temporal change of the calyx tube length in Clematis stans(Ranunculaceae). AB - Clematis stans is dioecious semi-arboreal, with pale purple-blue, nodding, tubulous flowers in a paniculate inflorescence. Both male and female flowers produce nectar from the base of the calyx tube during a flowering period of 3 or 4 days, and are pollinated by two bumblebee species, Bombus diversus and B. honshuensis,with different proboscis lengths. When the flowers open, four sepals constructing a calyx tube separate at the top and their respective tips gradually curl up, so that a tubular part shortens. Observations at two field sites showed that B. diversus (with a longer proboscis) most often visits the flowers with a longer calyx tube, and B. honshuensis (with a shorter proboscis) the flowers with a shorter calyx tube, i.e., later in the flowering period. By changing the calyx tube length, the flowers of C. stans accept the two bumblebee species with different proboscis length as pollinators and thus increase the chance of pollination for each flower. It was also found that the two bumblebee species prefer the male flowers to the female flowers, although the female flowers secrete more nectar as a reward than male flowers. This is likely because they visit the male flowers to collect pollen grains in addition to nectar. PMID- 12579361 TI - Ovules and seeds in Euphorbioideae (Euphorbiaceae): structure and systematic implications. AB - Ovule and seed structure in Euphorbioideae, one of the five euphorbiaceous subfamilies, is surveyed to evaluate its systematic implications on the basis of 79 species representing four of five tribes. All Euphorbioideae, like two other "uniovulate" subfamilies Acalyphoideae and Crotonoideae, but unlike most of two "biovulate" subfamilies Oldfieldioideae and Phyllanthoideae, consistently have a persistent and palisadal exotegmen composed of radially elongate, sclerotic, and pitted cells. Within Euphorbioideae, the tribe Stomatocalyceae (also with the palisadal exotegmen) is unusual in having vascular bundles in outer integument and clearly distinct from the remaining Euphorbioideae and the other "uniovulate" subfamilies. With the exclusion of Stomatocalyceae, Euphorbioideae are not anatomically divided into major groups such as a pseudanthial and a non pseudanthial clade, but instead have some remarkable diversity within a tribe, a subtribe, and even a genus in the three ovule and seed characters: (1) the thickness of the inner integument, (2) the thickness of the outer integument, and (3) the presence or absence of an aril. Groups of genera and species wrapped by different combinations of their characteristics, however, are not necessarily harmonized with tribal or subtribal classifications available. Anatomical similarities and dissimilarities presented in this paper, as well as relationships among taxa presented in the classifications available, will be critically evaluated in the light of results of ongoing molecular phylogenetic analyses. PMID- 12579362 TI - Cell division and cell enlargement in isolated Cucurbita cotyledons grown in darkness and in light. AB - The spatial and temporal patterns of post-embryonal cell growth and cell division were characterised in excised cotyledons of vegetable marrow ( Cucurbita pepo L. var. giromontia Alef.) incubated in water. The concurrent roles of these two processes in cotyledon growth were determined using paradermal sections of the first palisade layer of developing cotyledons. Tissue specificity was observed in the pattern of cell division. The daughter cells derived from an initial cell, which had already differentiated before imbibition of the seeds, were tightly packed in a cluster, which enabled us to monitor cell division during early cotyledon development. Heterogeneity of cell size was recognised during the process of cell proliferation in the cluster, suggesting that cell division is uncoupled from control of cell size. There was significantly more cell division in the marginal part of the cotyledons than in other parts, suggesting high activity of the marginal meristem. Light enhanced cell and cotyledon enlargement, but had no effect on the number of divisions. This study elucidated the cellular basis of post-germinative Cucurbita cotyledon morphogenesis and development. PMID- 12579363 TI - Phylogenetics of Miscanthus, Saccharum and related genera (Saccharinae, Andropogoneae, Poaceae) based on DNA sequences from ITS nuclear ribosomal DNA and plastid trnLintron and trnL-F intergenic spacers. AB - DNA sequences were used to assess the monophyly and inter-relationships of Miscanthus, Saccharumand related genera in the Saccharum complex. Three DNA regions were sequenced, including the trnLintron and the trnL-F intergenic spacer of the plastid genome and the ITS region of nuclear ribosomal DNA (nrDNA). Because it was more variable, the ITS region proved most suitable for phylogenetic reconstruction at this level, and the results indicate that Miscanthus s.l. and Saccharum s.l. are polyphyletic. A set of species from Saccharum section Ripidium(clade a) do not group closely with any members of Saccharum s.l. A number of Miscanthus species from eastern or south-eastern Asia represent a monophyletic group with a basic chromosome number of 19 (clade b), but the other species from Africa and the Himalayas are clearly excluded. There is support for a monophyletic Saccharum s.s. clade including S. officinarumand S. spontaneum that is sister to Miscanthus s.s(clade c). There is no evidence to support the division of some Saccharum s.l. into the genera currently known as Erianthus and Narenga. Saccharum contortum( =Erianthus contortus), S. narenga (= Narenga porphyrocoma) and Erianthus rockii, group more closely with Miscanthus fuscus, a species from the Himalayas and also with the African Miscanthus s.l. species (= Miscanthidium, clade d). PMID- 12579364 TI - Purification and properties of aminoaldehyde dehydrogenase from Avena sativa. AB - NAD-dependent aminoaldehyde dehydrogenase (AMADH, EC 1.2.1.-) from Avena shoots was purified by DEAE Sephacel, hydroxyapatite, 5'-AMP Sepharose 4B, Mono Q, and TSK-GEL column chromatographies to homogeneity by the criterion of native PAGE. SDS-PAGE yielded a single band at a molecular mass of 55 kDa. IEF studies showed a band with a p I value of 5.3. In contrast to AMADHs from other species, the TSK GEL chromatography showed that AvenaAMADH exists as a monomer in the native state. The purified enzyme catalyzed the oxidations of 3-aminopropionaldehyde (APAL), 4-aminobutyraldehyde (ABAL) N-(3-aminopropyl)-4-aminobutyraldehyde (APBAL), and 4-guanidinobutyraldehyde (GBAL), but not of betaine aldehyde or indoleacetaldehyde. The K(m) values for APAL, ABAL, and GBAL were 1.5x10(-6), 2.2x10(-6), and 1.3x10(-5) M, respectively. Although N-terminal amino acid sequence of Avena AMADH could not be determined due to a modification of the amino residue, the sequence of the fragment of AMADH cleaved by V8 protease showed greater similarity to the barley BADH than to the pea AMADH. PMID- 12579365 TI - A review of brood-site pollination mutualism: plants providing breeding sites for their pollinators. AB - In this paper, I review pollination systems in which plants provide breeding sites as a reward for pollination. I divide the pollinators into three groups based upon ovipositing sites and the larval food of insects. The first group consists of ovule parasites found in only five plant lineages, e.g., the fig wasps and yucca moths, pollination systems in which pollinator specificity is very high. The second group is pollen parasitism, primarily by thrips (Thysanoptera), but specificity of the pollinators is low. In the third group, pollinator larvae (Coleoptera and Diptera) develop in decomposed flowers and inflorescences of plants and these adaptations evolved repeatedly via different pathways in various plant taxa. Pollinator specificity varies, and shifts in pollinators may occur between related or unrelated insects. PMID- 12579366 TI - Regulation of the biosynthesis of plant hormones by cytochrome P450s. AB - Cytochrome P450 monooxygenases are a large group of heme-containing enzymes, most of which catalyze hydroxylation reactions. Since the discovery of cytochrome P450 in plants, more than 500 forms have been found, and they appear to be involved in the biosynthetic pathways of a large variety of primary and secondary metabolites. In particular, cytochrome P450s are involved in the biosynthesis of plant hormones, and play important roles in the regulation of plant growth and development. Recent genetic and functional analyses of cytochrome P450s in plants have significantly improved our understanding of not only the biosynthetic pathways themselves, but also of plant development from the perspective of hormonal control of morphogenesis. This review summarizes the present status of research on cytochrome P450s' roles in regulating the biosynthesis of plant hormones. PMID- 12579367 TI - Carbon and nitrogen contents of food bodies in three myrmecophytic species of Macaranga: implications for antiherbivore defense mechanisms. AB - In Macaranga myrmecophytes, differences in the production of the food bodies (FBs), on which symbiont ants feed, may relate to the intensity of antiherbivore defense by the ants. Interspecific comparisons among Macaranga species on such a mutualistic cost give important information on their strategies and evolution of antiherbivore defense. In this study, the carbon and nitrogen contents of FBs as well as the production rate of FBs were measured in three Macaranga species, M. winkleri, M. trachyphylla, and M. beccariana. There were significant differences in the production rates of FBs among species; the investment in FBs was greater in the Macaranga species in which ant defenses were more intensive. The carbon and nitrogen contents of FBs were significantly different among the three species, although they did not match the intensity of ant defense; the nitrogen content, especially, was greatest in the species of least intensive ant defense. It is suggested that Macaranga plants may have differentiated in the dependence on ant defense by controlling the total amount of nitrogen of FBs, not simply by nitrogen content. PMID- 12579368 TI - Differentiation in the status of self-incompatibility among Calibrachoa species (Solanaceae). AB - The overall status of self-incompatibility, as assessed by the rate of capsule set after self-pollination, was investigated in the genus Calibrachoa (Solanaceae). Thirty-two species were surveyed using a total of 655 individuals collected in 102 different native populations in Argentina, Brazil, Mexico, and Uruguay. The rate of capsule-set in 278 voucher specimens collected from the same native habitats was also measured to obtain additional information to assess the degree of self-(in)compatibility. Only one species, Calibrachoa parviflora, was self-compatible (SC, autogamous) and the other 31 species were found to be self incompatible (SI). A mixed population (SI and SC individuals in the same population) was not found. The differentiation of C. parviflora as an autogamous species is associated with a successful occupation of different (riparian) habitats within a larger range of geographic distribution compared to the rest of the species in the principally SI genus of Calibrachoa. PMID- 12579369 TI - Accelerated shoot overgrowth of rice mutant ao-1 is epistatic to gibberellin sensitive and -insensitive dwarf mutants. AB - A shoot overgrowth mutant of rice ( Oryza sativa L.), accelerated internode overgrowth-1 ( ao-1), is marked by accelerated longitudinal elongation of aerial parts and overgrowth of internodes at the vegetative stage. The physiological properties of ao-1 were similar to those of wild plants treated with a saturating level of exogenous gibberellins (GAs), except for the internode-overgrowth phenotype, which was not mimicked by GA-treated wild plants. The ao-1 mutant was less sensitive to a GA biosynthesis inhibitor, Uniconazole-P, than the wild type. Dwarf alleles of three loci, including two GA-sensitive and one GA-insensitive mutation, were introduced to produce double-mutants with ao-1, but the overgrowth phenotype was not suppressed in double-homozygous mutants. These results suggest that the overgrowth phenotype of ao-1 is caused by abolition of GA signaling rather than by GA overproduction. It is likely that a part of the shoot regulation system of ao-1 is saturated with the GA signal. As a possible model consistent with the results, we propose that AO-1 protein acts as a negative regulator in GA signal transduction. PMID- 12579370 TI - Genetic variation and population differentiation of Michelia formosana (Magnoliaceae) based on cpDNA variation and RAPD fingerprints: relevance to post Pleistocene recolonization. AB - We used sequence variation of the atpB- rbcL intergenic spacer of cpDNA and nested clade analysis to assess the phylogeographic pattern of Michelia formosana, a species restricted to Taiwan and the Ryukyus. In total, 31 haplotypes were identified and clustered into four major chlorotypes. Genetic composition of nearly all populations was heterogeneous and paraphyletic phylogenetically. Although the apportionment of cpDNA variation hardly revealed a geographic pattern due to the coancestry of dominant sequences, some chlorotypes were restrictedly distributed. According to the patterns of clade dispersion and displacement, a reconstructed minimum spanning network revealed that historical events of past fragmentation and range expansion, associated with glaciation, may have shaped the phylogeographic patterns of M. formosana. Four possible refugia were identified: the Iriomote and Ishigaki Islands (the southern Ryukyus), Wulai (northern Taiwan), and Nanjen (southern Taiwan), on the basis of the interior positions of their haplotypes in the network and the high level of nucleotide diversity. Given insufficient time for coalescence at the cpDNA locus since the late Pleistocene recolonization, lineage sorting led to low levels of genetic differentiation among populations. In contrast, hierarchical examination of the random amplified polymorphic DNA (RAPD) data scored from six populations across three geographical regions, using an analysis of molecular variance (AMOVA), indicated high genetic differentiation both among populations (Phi(ST) = 0.471) and among regions (Phi(CT) = 0.368). An unweighted pair group method with arithmetic mean (UPGMA) tree of the RAPD fingerprints revealed that populations of two offshore islands of eastern Taiwan ( M. formosana var. kotoensis) were clustered with geographically remote populations of the Ryukyus instead of those in southern Taiwan, suggesting some historical division due to geographic barriers of the central mountain range. In contrast to the paraphyly of the nearly neutral cpDNA alleles, differentiated RAPDs may have experienced diversifying selection. PMID- 12579371 TI - Molecular evidence for the taxonomic identity of Korean Adonis (Ranunculaceae). AB - Although Korean Adonis has been traditionally recognized as Adonis amurensis Regel and Radde with various infraspecific taxa described, its taxonomic identity is still in dispute. We investigated the genetic variation in 60 individuals from 12 populations in Korea to elucidate the taxonomic identity of the Korean Adonis complex. Random amplified polymorphic DNA (RAPD) analysis revealed that Korean Adonis comprises three species: A. amurensis, A. pseudoamurensis, and A. multiflora. Adonis amurensis is distributed in northern central inland regions of Korea and A. pseudoamurensis is found in southern parts of the Korean peninsula. Adonis multiflora grows only on Cheju Island, which is the southernmost part of Korea. Phylogenetic analysis of nuclear ribosomal internal transcribed spacer (ITS) sequences partially supported the presence of three Adonis taxa in Korea as detected by RAPD analysis. The Adonis population on Jangbong Island located in the West Sea, which was referred to as A. pseudoamurensison the basis of morphological examination, was separated from the other populations of A. pseudoamurensis. Otherwise, the molecular evidence is well congruent with the recent morphological study that proposes that Korean Adonis consists of these three species. PMID- 12579372 TI - Karyomorphology of Taiwanese Begonia (Begoniaceae): taxonomic implications. AB - The karyomorphology of all 14 species of Taiwanese Begonia was investigated to elucidate their chromosome features and chromosomal evolution. Among all species investigated, differences in chromosome features are found in: (1) chromosome number 2 n = 22, 26, 36, 38, 52, 60, 64, 82, and (2) frequencies of chromosomes with secondary, tertiary, and/or small constrictions of polyploids, ranging from 23% to 63%, which is higher than the expected value of about 9%. It is suggested that after polyploidization from the diploid species (i.e., 2 n = 22 and frequencies of chromosomes with secondary, tertiary, and/or small constrictions of polyploids of about 9%), chromosome translocations occurred, followed by a decrease in chromosome number, and subsequently stabilized genomes were formed in various species in Taiwan. The karyomorphological evidence also suggested that the chromosome morphology has evolved in parallel in the begonias belonging to different sections in Taiwan. The variation in chromosomal features is more complex than the variation in floral and fruit morphologies. Karyomorphological data also supports the recognition of five new species in Taiwan: Begonia bouffordii, B. chuyunshanensis, B. pinglinensis, B. tengchiana, and B. wutaiana. Based on detailed karyomorphological analyses, the taxonomic implications, speciation, and chromosomal evolution in Taiwanese Begoniaare discussed. PMID- 12579373 TI - MAP kinase cascades in elicitor signal transduction. AB - Protein kinases play important roles in elicitor signal transduction. In this article, I describe the current view of the role of mitogen-activated protein kinase (MAPK) cascades in elicitor signal transduction of plant cells based on our own research and recent developments in this field. In the past several years, it has become apparent that MAPK cascades play important roles in elicitor signal transduction in plants. Our early studies demonstrated the identification of p47 MAPK in tobacco as an elicitor-responsive protein kinase and possible involvement of p47 MAPK in elicitor signal transduction to induce defense responses, including defense gene expression and hypersensitive cell death. However, the molecular identity of p47 MAPK is still unclear. Recent important studies suggest that tobacco MAPK cascades that include SIPK, and/or WIPK, and NtMEK2, an upstream kinase for both SIPK and WIPK, have a crucial function in induction of defense responses and hypersensitive cell death. The orthologs of these protein kinases in Arabidopsis and alfalfa are also suggested to have similar functions. Furthermore, the identification of loss-of-function mutation in Arabidopsis reveals a negative regulatory role for putative MAPK cascades in plant defense mechanisms. PMID- 12579374 TI - Differential expression of genes influenced by changing salinity using RNA arbitrarily primed PCR in the archaeal halophile Haloferax volcanii. AB - Extreme halophiles belonging to the domain Archaea require a minimum of approximately 10% NaCl for growth. Many of these obligate halophiles will continue to grow even as NaCl concentrations approach saturation. The haloarchaeon Haloferax volcanii is a model organism in which to study the effects of changes in medium salinity on gene expression, as this organism grows over a wide range of NaCl concentrations, between 12% and 23%, with little effect on growth rate. An RNA arbitrarily primed PCR (RAP-PCR) approach has been applied to identify those genes that are differentially expressed in response to changing salinity. Differences in gene expression can be detected using this methodology, as each sample generates its own unique RNA fingerprint for each growth condition examined. RNA was prepared from H. volcanii cultures grown with two different NaCl concentrations in the medium, RAP-PCR was performed, and seven differentially expressed transcripts were identified. These fragments were cloned, sequenced, and subjected to transcript analysis to confirm their regulation. One of the sequences identified in this study displays homology to the eukaryotic Ser/Thr protein kinase Ire1p, a sensor of protein unfolding in yeast and mammalian cells. Evidence for serine phosphorylation in H. volcanii is also presented. PMID- 12579375 TI - Citrate synthase from Thermus aquaticus: a thermostable bacterial enzyme with a five-membered inter-subunit ionic network. AB - A bacterial thermostable citrate synthase has been analyzed to investigate the structural basis of its thermostability, and to compare such features with those previously identified in archaeal citrate synthases. The gene encoding the citrate synthase from Thermus aquaticus was identified from a gene library by screening with a PCR fragment amplified from genomic DNA using a primer based on the determined N-terminal amino acid sequence and a citrate synthase consensus primer. Apart from high sequence similarities with citrate synthase sequences within the Thermus/ Deinococcus group, the analyzed enzyme has highest similarities with the enzyme from the hyperthermophilic Archaeon Pyrococcus furiosus. The recombinant enzyme is a dimer with high specific activity. Compared to its thermoactivity (T(opt)at 80 degrees C), the thermal stability of the enzyme is high, as judged from its T(m) (101 degrees C), and from irreversible thermal inactivation assays. Molecular modeling of the structure revealed an inter-subunit ion-pair network, comparable in size to the network found in the citrate synthase from P. furiosus; these networks are discussed in relation to the high thermal stability of these bacterial and archaeal enzymes. PMID- 12579376 TI - GroEL from the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC 125: molecular characterization and gene cloning. AB - The heat shock response of the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC 125 (PhTAC 125) gives rise to the production of several inducible proteins. Among these, the protein corresponding to a 55-kDa band on SDS-PAGE was purified to homogeneity and identified as a GroEL-like protein. The gene coding for this protein (PhGroEL) was cloned and sequenced; the deduced amino acid sequence shows 82% sequence identity to GroEL from Escherichia coli (EcGroEL). The ORF found in the 5' upstream region codes for a homologue of the GroES from E. coli (PhGroES, 71% sequence identity to EcGroES). PhGroEL shows a chaperone activity and can use GroES from E. coli as a co-chaperone. PhGroEL melting temperature, 6 degrees C lower than that of EcGroEL, and equilibrium unfolding experiments in urea showed a less stable protein architecture for the psychrophilic GroEL. The data herein reported demonstrate that PhGroEL cold adaptation consists in a shift of the protein properties toward lower temperatures without increasing catalytic efficiency at low temperatures. Primary extension analysis depicted a complex organization of regulative elements for the operon containing the genes coding for PhgroES and PhgroEL (PhgroE), suggesting that a fine-tuning of transcription can also be involved in thermal adaptation of PhTAC 125. PMID- 12579377 TI - Salinisphaera shabanensis gen. nov., sp. nov., a novel, moderately halophilic bacterium from the brine-seawater interface of the Shaban Deep, Red Sea. AB - A novel, moderately halophilic bacterium was isolated from the brine-seawater interface of the Shaban Deep, northern Red Sea. A polyphasic approach was used for the taxonomic characterization of this isolate, with the phenotypic and phylogenetic data clearly showing the distinctiveness of this bacterium. Cells of isolate E1L3A were Gram-negative, monotrichous cocci that showed a remarkable physiological flexibility, as could be seen by the quite broad growth ranges for oxygen, temperature, NaCl, and, to a smaller degree, pH. In addition, it was able to grow from atmospheric pressure up to 15 MPa, making it a piezotolerant bacterium. Phylogenetically, strain E1L3A represents a new, deeply branching lineage within the gamma-Proteobacteria, as determined by 16S rRNA gene sequence analysis. No close relatives are known so far, with sequence similarity to other cultivated members of the gamma-Proteobacteria being lower than 88%. The creation of the new genus Salinisphaera and the new species Salinisphaera shabanensis (DSM 14853; JCM 11575) for this new and highly versatile microorganism is therefore proposed. PMID- 12579378 TI - Purification and characterization of Thermus thermophilus UvrD. AB - The DNA helicase UvrD (helicase II) protein plays an important role in nucleotide excision repair, mismatch repair, rolling circular plasmid replication, and in DNA replication. A homologue of the Escherichia coli uvrD gene was previously identified in Thermus thermophilus; however, to date, a UvrD helicase has not been purified and characterized from a thermophile. Here we report the purification and characterization of a UvrD protein from Thermus thermophilus HB8. The purified UvrD has a temperature range from 10 degrees to >65 degrees C, with an optimum of 50 degrees C, within the temperature limits of the assay. The enzyme had a requirement for divalent metal ions and nucleoside triphosphates which related to enzyme activity in the order ATP > dATP > dGTP > GTP >> CTP > dCTP >> UTP. A simple real-time helicase assay was developed that should facilitate detailed kinetic studies of the enzyme. Evaluation of helicase substrates using this assay showed that the enzyme was highly active on a double stranded DNA with 5' recessed ends in comparison with substrates with 3' recessed or blunt ends, and supports enzyme translocation in a 3'-5' direction relative to the strand bound by the enzyme. PMID- 12579379 TI - Purification and characterization of a new hyperthermostable, allosamidin insensitive and denaturation-resistant chitinase from the hyperthermophilic archaeon Thermococcus chitonophagus. AB - A new chitinase (1,4-beta-D-N-acetyl-glucosaminidase, EC 3.2.1.14) was detected and purified to homogeneity in its native form from the chitinolytic enzyme system of the extremely thermophilic archaeon Thermococcus chitonophagus. This is the first nonrecombinant chitinase purified and characterized from archaea and also constitutes the first case of a membrane-associated chitinase isolated from archaea. The enzyme is a monomer with an apparent molecular weight of 70 kDa [therefore named chitinase 70 (Chi70)] and pI of 5.9; it is hydrophobic and appears to be associated with the outer side of the cell membrane. Chi70 is optimally active at 70 degrees C and pH 7.0 and exhibits remarkable thermostability, maintaining 50% activity even after 1 h at 120 degrees C, and therefore the enzyme is the most thermostable chitinase so far isolated. The enzyme was not inhibited by allosamidin, the natural inhibitor of chitinolytic activity, and was also resistant to denaturation by urea and SDS. On the other hand, guanidine hydrochloride significantly reduced enzymatic activity, indicating that, apart from the hydrophobic interactions, ion pairs located on the surface of the protein could be playing an important role in maintaining the protein's fold and enzyme activity. Chi70 showed broad substrate specificity for several chitinous substrates and derivatives. The lowest K(m) and highest K(cat) values were found for pNP(NAG)(2) as substrate and were determined to be 0.14 mM and 23 min(-1), respectively. The hydrolysis pattern was similar for oligomers and polymers, with N, N'-diacetylchitobiose [(NAG)(2)] being the final, major hydrolysis product. Chi70 was classified as an endochitinase due to its ability to release chitobiose from colloidal chitin. Additionally, the enzyme presented considerable cellulolytic activity. Analysis of the NH(2)-terminal amino acid sequence showed no detectable homology with other known sequences, suggesting that Chi70 is a new protein. PMID- 12579380 TI - Halomonas glaciei sp. nov. isolated from fast ice of Adelie Land, Antarctica. AB - Eleven psychrophilic bacteria were isolated from a solid layer of fast ice in the middle of Pointe-Geologie Archipelago, Adelie Land, Antarctica. The 11 isolates based on the phenotypic characteristics, chemotaxonomic and phylogenetic analysis have been identified as members of the genus Halomonas. All the isolates at the 16S rDNA sequence level were identical, possessed the 15 conserved nucleotides of the family Halomonadaceae and four nucleotides of the genus Halomonas. Therefore, the 16S rDNA sequence of DD 39 was used for calculating the evolutionary distances and for phylogenetic analysis. It was observed that DD 39 formed a robust cluster with H. variabilis, from which it differed by 0.7%. Further DNA DNA hybridization studies indicated low DNA-DNA homology (15%) between H. variabilis and DD 39. Between the 11 Antarctic isolates the homology was >85%. In addition it was observed that DD 39 was different from H. variabilis in that it was psychrophilic, could tolerate only up to 15% sodium chloride, could not hydrolyse esculin, could not reduce nitrate, was urease negative, could not utilize glycerol as a carbon source, and was resistant to ampicillin and erythromycin and sensitive to nalidixic acid. In addition, it also exhibited distinct differences with respect to high content of C(16:1) and low levels of cyclo-C(17:0) and cyclo-C(19:0). DD 39 also differed from all the other reported species of Halomonas with respect to many phenotypic characteristics. It is proposed therefore that DD 39 should be placed in the genus Halomonas as a new species that is Halomonas glaciei. The type strain of H. glaciei is DD 39(T) (MTCC 4321; JCM 11692). PMID- 12579381 TI - A gene encoding a novel extremely thermostable 1,4-beta-xylanase isolated directly from an environmental DNA sample. AB - Small-subunit (SSU) rRNA genes (rDNA) were amplified by PCR from a hot pool environmental DNA sample using Bacteria- or Archaea-specific rDNA primers. Unique rDNA types were identified by restriction fragment length polymorphism (RFLP) analysis and representative sequences were determined. Family 10 glycoside hydrolase consensus PCR primers were used to explore the occurrence and diversity of xylanase genes in the hot pool environmental DNA sample. Partial sequences for three different xylanases were obtained and genomic walking PCR (GWPCR), in combination with nested primer pairs, was used to obtained a unique 1,741-bp nucleotide sequence. Analysis of this sequence identified a putative XynA protein encoded by the xynA open reading frame. The single module novel xylanase shared sequence similarity to the family 10 glycoside hydrolases. The purified recombinant enzyme, XynA expressed in E. coli exhibited optimum activity at 100 degrees C and pH 6.0, and was extremely thermostable at 90 degrees C. The enzyme showed high specificity toward different xylans and xylooligosaccharides. PMID- 12579382 TI - Isolation of fusion proteins containing SecY and SecE, components of the protein translocation complex from the halophilic archaeon Haloferax volcanii. AB - By exploiting the salt-insensitive interaction of the cellulose-binding domain (CBD) of the Clostridium thermocellum cellulosome with cellulose, purification of CBD-fused versions of SecY and SecE, components of the translocation apparatus of the halophilic archaeon Haloferax volcanii, was undertaken. Following transformation of Haloferax volcanii cells with CBD-SecY- or -SecE-encoding plasmids, cellulose-based purification led to the capture of stably expressed, membrane-bound 68 and 25 kDa proteins, respectively. Both fusion proteins were recognized by antibodies raised against the CBD. Thus, CBD-cellulose interactions can be employed as a salt-insensitive affinity purification system for the capture of complexes containing the Haloferax volcanii translocation apparatus components SecY and SecE. PMID- 12579383 TI - Multi-subunit assembly of the Pyrococcus furiosus small heat shock protein is essential for cellular protection at high temperature. AB - The hyperthermophilic archaeon, Pyrococcus furiosus, expresses a small, alpha crystallin-like protein in response to exposure to extreme temperatures, above 103 degrees C. The P. furiosus small heat shock protein (Pfu-sHSP) forms large oligomeric complexes. Based on the available crystal structures of the Methanocaldococcus jannaschii and wheat sHSPs, the protruding carboxy terminal domain is probably involved in subunit interactions. We constructed Pfu-sHSP mutants to analyze chaperone function and to study multi-subunit assembly. The results confirmed that the carboxy terminus of Pfu-sHSP is involved in inter dimer interactions, whereas the amino terminal deletion mutant still exhibited the wild-type assembly characteristics. The ability to form oligomeric complexes via the carboxy terminal domain was shown to be necessary for thermotolerance of Escherichia coli overexpressing Pfu-sHSP. The amino terminal domain was not required for inter-species thermotolerance. PMID- 12579384 TI - [Preemptive randomized, double-blind study with lornoxicam in gynecological surgery]. AB - AIM: Lornoxicam is a non opioid analgesic belonging to the oxicam group. The aim of this study was to determine whether lornoxicam has a preemptive analgesic effect. METHODS: This study was carried out in a randomized, double-blind fashion with 66 patients divided into three groups undergoing gynecological operations. Group I was administered 8 mg of lornoxicam i.v. preoperatively followed by an 8 mg bolus every 8 h for a total dose of 24 mg in the first 24 h. Group II was administered 8 mg of lornoxicam i.v. bolus before the end of the operation followed by 8 mg every 8 h for a total dose of 24 mg in the first 24 h. Group III was administered placebo before and after the operation and for the first 24 h. The effectiveness was assessed postoperatively using the visual analogue scale (at rest, on exertion) and by calculating the total analgesic consumption of morphine hydrochloride in the first 24 h following operation. Vital signs and side effects were documented. RESULTS: Groups I and II demonstrated significantly reduced pain scores compared to group III at various points in time. Group I also demonstrated a weakly significant reduction in analgesic consumption of morphine hydrochloride postoperatively compared to groups II and III. CONCLUSION: Lornoxicam administered preemptively appears to improve the quality of postoperative analgesia and lead to reduced consumption of opioid analgesics postoperatively in patients undergoing gynecological operations. PMID- 12579385 TI - [Prophylaxis of phantom pain: is regional analgesia ineffective?]. AB - AIM: A recently published randomized study failed to show a significant reduction of phantom limb pain after perioperative epidural analgesia EDA [9]. Since these findings were not supported by previous studies,we conducted an analysis of factors contributing to the results of phantom limb pain prophylaxis. We calculated the efficacy of perioperative EDA as "Number Needed to Treat" (NNT). METHODS: Included studies were retrieved by a medline-search from 1966 to 1999 and published articles on phantom limb pain prophylaxis. First,we analyzed the influence of patient data, type of intervention and study design on the results. Then,we stratified studies in comparable groups and outcome measures. The efficacy of perioperative EDA in phantom limb pain prophylaxis was calculated using NNT's. The outcome criterion in this analysis was "free of phantom limb pain < or = 3 on an analogue scale from 0 ( no pain) to 10 (worst pain) 12 months after amputation". RESULTS: Variations in preoperative pain, start and duration of regional analgesia and the definition of phantom limb pain were associated with different results. After stratifying the results by the above mentioned definition of phantom pain intensity, only time and duration of regional analgesia showed effects on the study results. Pre-, intra- and postoperative EDA was associated with a significant reduction of phantom limb pain 12 months after amputation, NNT = 5,8 (95%-CI 3,2-28,6). However, a reduction of phantom limb pain by postoperative EDA alone could not be confirmed on the basis of the analysed data. CONCLUSIONS: Perioperative EDA has been shown to be an effective prophylaxis of phantom limb pain. The most important differences between studies were the definition of phantom limb pain by intensity ratings. Thus, perioperative EDA does not completely abolish phantom limb pain, but increases the number of patients with a mild form of phantom pain. PMID- 12579386 TI - [Pain specialist in the assessment process]. AB - Pain specialists are often involved in assessing social medical aspects of chronic pain patients. A standardized examination in the assessment process is not published. Data and statements on the assessment process between 1993 and 1997 were recorded from 15 experts by questionnaire. Statements to 568 reports were made. The majority of performance impairments were caused by back pain (35.9%) and headache (19.3%). All examiners integrated a psychological assessment. Although the assessment processes were very different, all examiners fulfilled the minimal criteria of the presented process for assessment of chronic pain patients. Verifying the symptoms as well as capturing some aspects of aggravation proved to be problematic. The present assessment practice is discussed with regard to the unequal procedure. On basis of the received data and the own experience in examination the process of structured assessment of chronic pain patients is presented under medical and psychological aspects. It contains a recommendation of a standardized process with minimal criteria and an optional supplement of the standardized process with problematical patients (e.g.demonstration, aggravation, simulation). Minimal demands on a qualified assessment of chronic pain patients should contain the study of the history, the German Pain-Questionnaire, the analysis of all the painful complaints with weighting their importance, the examination and the basic registration of sensational, emotional, cognitive, behavioral and social aspects of the chronic pain syndrome. An appended glossary of the most usable psychological tests with comments on their validation criteria aims to provide a more standard multimodal assessment of chronic pain patients complaints and functionability. PMID- 12579387 TI - [Manual medicine in chronic pain -- evaluation of an inpatient medical concept]. AB - OBJECTIVE: The influence of an inpatient manual medical concept for patients suffering from chronic muscular skeletal pain syndromes on the quality of life and on target parameters of a biopsychological and social concept of pain (VAS, PDI, CES-D) was investigated. Furthermore, it was looked at the predictability of treatment outcome according to the Mainz Pain Stages. METHOD: 211 patients of the Clinic for Manual Medicine were included in the study. Selected parameters such as pain intensity (VAS), pain distribution, disability in daily life (PDI), quality of life (SF-36) and grade of depression (CES-D) were evaluated on admission, discharge and 6 weeks after treatment. The patients were divided into groups according to the Mainz Pain Stages and compared to patients on the hospital waiting list (control group). RESULTS: The patients were well distributed within the Mainz Pain Stages. It was shown that the manual therapy concept influenced different parameters dependent on the Main Pain Stages.For Stage 1 and 2 the deduction in pain intensity was greatest, while for stage 3 the reduction in disability (PDI) was the most significant outcome. CONCLUSION: The therapeutic target should depend on the chronicity of the pain syndrome. Early interdisciplinary intervention, especially after increase in pain distribution is necessary to minimize chronicity. The Mainz Pain Stages are useful for the prediction of treatment outcome but still need further investigation for this group of patients. PMID- 12579388 TI - [Standardization of invasive neuromodulatory procedures]. AB - The Working Group on Neuromodulation of the German Association for the Study of Pain, composed of representatives from various scientific specialty societies, met on December 9, 2000, March 24, 2001, October 5, 2001, and December 8, 2001. As a result of these discussions grounded in current knowledge, the following guidelines were formulated for the standardization of invasive techniques of neuromodulation intended to serve as a systematic aid in decision-making and to provide recommendations for practice-oriented methods. The guidelines were based on both the clinical and practical experience of the group participants (see information box on the next page) as well as on the current scientific literature and guidance from the consensus report of the European Federation of IASP Chapters (EFIC) [23]. The guidelines serve the purpose of orientation and have no effect on either assumption of liability or discharge from liability. The guidelines were conceived for use by physicians in private practice,doctors in hospitals,and nonmedical personnel concerned with the care of chronic pain patients. The Working Group consists of unsalaried volunteers. The participants received no honorarium and were only reimbursed for normal travel expenses in accordance with customary directives. The guidelines will be revised should new scientific results become available, at the latest in 2 years. The plan exists to further develop the guidelines to stages II and III (AWMF). The Steering Committee of the DGSS appraised the guidelines and authorized the guidelines before publication. PMID- 12579389 TI - [Prevalence and risk factors of psychological burden and mental disorders in patients with musculoskeletal diseases -- a review of empirical studies]. AB - A musculoskeletal disease frequently entails numerous physical, social and occupational limitations, e.g. reduction of physical capabilities, chronic pain, and loss of employment. A number of patients suffer from comorbid psychological strain or mental disorders, and it frequently remains unclear whether these psychological phenomena are unconnected to, a consequence of or even among the causes for the somatic disease. In order to diagnose, or possibly prevent, comorbid mental disorders, it is necessary to know risk factors which are associated with mental disorders. Studies published during the last twenty years are reviewed which examine sociodemographic, disease specific and psychosocial factors in connection with psychological strain or mental disorders among patients with musculoskeletal diseases. Due to the relatively small number of relevant studies (n = 12), the findings have to be interpreted carefully. PMID- 12579390 TI - [Correlation between intrathecal opioids and gynecomastia -- a case report]. AB - INTRODUCTION: The intrathecal application of opioids is promoted as a safe and cost-efficient method to treat chronic pain of nonmalignant origin. But the way of application could trigger the appearance of otherwise rare side-effects. One of those side-effects could be the alteration of androgen hormones. Can a long time-application of intrathecal opioids result in gynecomastia? METHODS: Case report of a 49 year old with chronic back and limb pain. Research for publications. RESULTS: The close link between the begin of intrathecal application of opioids and the onset of gynecomastia in this patient refers to a causal correlation. Historic publications and new findings from animal research are supporting our hypothesis. CONCLUSIONS: Further research is needed to support the clinical suspicion of a correlation between intrathecal opioids and gynecomastia. In case of a opioid-produced gynecomastia tests of the oestrogen testosterone-ratio should be performed. An early substitution of testosterone could prevent a full fledged gynecomastia. PMID- 12579391 TI - [Therapy and prophylaxis of facial neuralgias and other forms of facial pain syndromes -- revised recommendations of the German Society of Migraine and Headache]. AB - Trigeminal neuralgia and postherpetic neuralgia are the most relevant neuralgiform facial pain syndromes. Trigeminal neuralgia is characterized by lancinating intensive pain attacks of very short duration, triggered by external cues,whereas postherpetic neuralgia consists predominantly of long-lasting burning pain. Sodium channel blocking drugs are first choice in treatment of trigeminal neuralgia, operative procedures encompass microvascular decompression,thermocoagulation and percutaneous retrogasserian glycerol rhizotomy. In the acute stage postherpetic neuralgia is treated antivirally and analgesically, in the chronic stage by tricyclic antidepressive substances. Other pain syndromes described encompass the Tolosa-Hunt-syndrome, cervicogenic headache, craniomandibular dysfunction syndrome, atypical facial pain and rarer syndromes. Therapeutic recommendations are based on evidence based medicine criteria (EBM). PMID- 12579392 TI - Diabetic nephropathy: renal development gone awry? AB - Nephrogenesis is controlled by a sequence of inductive signals between different areas of the developing kidney. As these signals are being elucidated, it has become clear that many important developmental genes are re-expressed in the mature organ following injury, possibly as part of repair and regeneration. While this reuse of developmental pathways may contribute to healing and repair, it may alternatively result in scar formation if specific components of the pathways are missing, if the temporal correlation of various elements is faulty, or if an injurious stimulus persists. In the review we will use diabetic nephropathy as an example to illustrate this paradigm in renal disease. The pathogenesis of diabetic nephropathy is complex and characterized by altered expression of many genes, including growth factors, apoptotic regulators, cellular matrix components, and cytoskeletal proteins. Many of these factors also function during kidney development. The elucidation of the roles these genes play in nephrogenesis and of their array of molecular partners and modulators may ultimately shed light on the pathogenesis of disease (and indeed vice versa), and may even suggest new therapeutic strategies. PMID- 12579393 TI - Developmental expression of HSP-72 and ischemic tolerance of the immature kidney. AB - The resistance of the immature kidney to ischemic injury is well documented, but the mechanisms involved in this tolerance have been elusive. Previous studies have demonstrated that tubules obtained from immature rats exhibit a bigger stress response than mature tubules. Consequently, we evaluated the developmental expression of HSP-72 in the postnatal kidney and determined whether or not that pattern of expression was correlated with the previously known tolerance of the immature kidney to injury. A distinct pattern of HSP-72 expression with a peak abundance at postnatal day 10 (P10), with a subsequent decline toward values seen in mature rats, was found. Moreover, this stress protein is located predominantly in tubular segments, the site of ischemic injury. To determine if this constitutive, non-induced expression of HSP-72 in the immature rat could be protective of cellular integrity and renal function, both immature (P10) and mature (8 weeks) rats were subjected to 45 min of bilateral renal artery ischemia. The postischemic induction of HSP-72 in the P10 animals was robust and the peak expression 2 h after ischemia was even greater than that detected in mature animals. Thus, the constitutive enhanced expression of HSP-72 did not prohibit or mute the inducible response of this stress protein in the immature animals. Immature animals, when compared with mature rats, also experienced cytoprotection, demonstrated by decreased detachment of Na-/K-ATPase from the cytoskeleton and substantial protection of renal function determined by serum creatinine level. These findings suggest that the developmental expression of heat shock proteins may play a critical and fundamental role in the well-observed tolerance of immature tubules to ischemic or anoxic injury. PMID- 12579394 TI - Response to Shiga toxin 1 and 2 in a baboon model of hemolytic uremic syndrome. AB - Post-diarrheal (D+) hemolytic uremic syndrome (HUS) is caused by Shiga-toxin (Stx)-producing Escherichia coli. There is epidemiological, cell culture, and mouse model evidence that Stx2-producing E. coli are more likely to cause HUS than strains that produce only Stx1, but this hypothesis has not been tested in a primate model of HUS. We have developed a baboon model of Stx-mediated HUS that was employed to compare the clinical, cytokine, and histological response to equal amounts of the two Shiga toxins. Animals given IV Stx2 developed progressive thrombocytopenia, hemolytic anemia, and azotemia, and urinary interleukin-6 levels rose significantly. Glomerular thrombotic microangiopathy was found at necropsy. Animals given Stx1 showed no cytokine response and no clinical, laboratory, or histological signs of HUS. Our findings from the primate model corroborate previous epidemiological, cell culture, and mouse model observations, and suggest that enteric infection with Stx2-producing E. coli is more likely to cause HUS than infection with organisms that produce only Stx1. PMID- 12579395 TI - Urinary heat shock protein-72 excretion in clinical and experimental renal ischemia. AB - Renal ischemia not only causes injury but also induces repair mechanisms, such as the cellular induction of the 72-kilodalton heat shock protein HSP-72. The aim of this study was to determine whether HSP-72 is excreted in urine after ischemic renal injury. The first urine of six pediatric allograft recipients was examined for proteinuria and urinary HSP-72 excretion. Sprague-Dawley rats were treated with renal ischemia or hyperthermia and renal cortex and urinary HSP-72 levels were determined. HSP-72 was excreted in the first urine of renal allografts. In rats, renal HSP-72 was induced both by renal ischemia or hyperthermia. However, only renal ischemia resulted in urinary excretion of HSP-72. Urinary excretion of HSP-72 indicates an increased renal stress response and loss of tubular cell integrity after clinical and experimental renal ischemia. PMID- 12579397 TI - Confirmation of the ATP6B1 gene as responsible for distal renal tubular acidosis. AB - Primary distal renal tubular acidosis (dRTA) type I is a hereditary renal tubular disorder, which is characterized by impaired renal acid secretion resulting in metabolic acidosis. Clinical symptoms are nephrocalcinosis, nephrolithiasis, osteomalacia, and growth retardation. Biochemical alterations consist of hyperchloremic metabolic acidosis, hypokalemia with muscle weakness, hypercalciuria, and inappropriately raised urinary pH. Autosomal dominant and rare forms of recessive dRTA are known to be caused by mutations in the gene for the anion exchanger AE1. In order to identify a gene responsible for recessive dRTA, we performed a total genome scan with 303 polymorphic microsatellite markers in six consanguineous families with recessive dRTA from Turkey. In four of these there was an association with sensorineural deafness. The total genome scan yielded regions of homozygosity by descent in all six families on chromosomes 1, 2, and 10 as positional candidate region. In one of these regions the gene ATP6B1for the ss1 subunit of the vacuolar H(+)-ATPase is localized, which has recently been identified as causative for recessive dRTA with sensorineural deafness. Therefore, we conducted mutational analysis in 15 families and identified potential loss-of-function mutations in ATP6B1in 8. We thus confirmed that defects in this gene are responsible for recessive dRTA with sensorineural deafness. PMID- 12579396 TI - Comparison of mycophenolate mofetil and azathioprine in obstructive nephropathy. AB - The effect of mycophenolate mofetil (MMF) and azathioprine (AZA) in reducing renal interstitial fibrosis in rats with unilateral ureteral obstruction (UUO) was studied. Fifty-nine rats with surgically induced UUO received oral MMF (n=19), AZA (n=19), or no treatment (n=21). The obstructed kidneys were analyzed by histology and morphometry on days 21 and 60 post UUO. Fibronectin- and collagen-stained areas were significantly lower in both treatment groups when compared with the control group 21 days post surgery. Transforming growth factor (TGF)-beta expression was significantly lower in the MMF-treated group than in the AZA-treated (P<0.01) and the control (P<0.001) groups. There was no significant difference in TGF-beta expression between the AZA-treated and the control groups 21 days post surgery (P>0.05). No significant difference was found in TGF-beta and fibronectin expression between treated and untreated groups on the 60th day post surgery. However, collagen expression was significantly lower in both treated groups than in the untreated group on the 60th day (P<0.005). We observed that MMF is more effective in preventing fibrosis than AZA in the UUO model in the short term; however, there is a less significant anti-fibrotic effect of these drugs in long-term than in short-term obstruction. PMID- 12579399 TI - Early treatment of urinary infection prevents renal damage on cortical scintigraphy. AB - Diagnosis of urinary infection in young children is often delayed, which may result in renal damage. However, it remains to be clarified how soon the treatment should be started to prevent renal changes. The present study prospectively enrolled young children with diagnosis of their first febrile urinary infection who underwent technetium-99m dimercaptosuccinate renal cortical scintigraphy within 120 h of initiation of treatment. Patients with abnormal renoscintigraphy received antibiotics for 2 weeks and scintigraphy was repeated 1 year later. Twenty-two children were enrolled from July 1995 through March 2000. Acute-phase renoscintigraphy identified focal defects in 0 of the 14 children who were treated within 24 h of the disease, 1 of the 3 treated in 24-48 h, and 2 of the 5 treated in 48-72 h. Repeat renoscintigraphy showed disappearance of the focal defects in all 3 children. The present study has shown that early treatment within 24 h of onset of the fever due to urinary infection should deter renal changes. Fever for more than 24 h prior to diagnosis indicates a high risk for renal changes and needs an immediate effective treatment to avoid renal damage. PMID- 12579398 TI - Renin-angiotensin system polymorphisms and renal scarring. AB - The purpose of the study was to determine whether DNA polymorphisms at the renin angiotensin-aldosterone (RAS) genes were associated with evolution to renal scar formation and, consequently, with reflux nephropathy (RN) in patients with vesicoureteral reflux (VUR). Some authors have suggested that the DD genotype of the angiotensin-converting enzyme (ACE) gene would be an adverse renal prognosis factor. We recruited 246 patients (aged 3 months to 22 years) from four Spanish hospitals. These included 69 patients with VUR, 110 with RN (determined by absence/presence of renal scarring on dimercaptosuccinc acid scan), 27 with chronic renal failure due to RN, and 40 patients (control group) with urinary tract infection and normal findings on renal ultrasonography and voiding cystoureterogram. The ACE I/D, angiotensin II type 1 receptor AT1 A1166C, angiotensin II type 2 receptor A3123C AT2, and angiotensinogen AGT M235T polymorphisms were determined on the basis of polymerase chain reaction amplification. ACE serum levels were determined by spectrophotometric methods. We found no statistical differences in the distribution of RAS polymorphisms between the different groups. The ACE D allele was linked to higher ACE serum levels. We found no association between ACE I/D polymorphism and presence of hypertension, proteinuria, grade of VUR, or unilateral/bilateral VUR. Patients with the DD genotype had a lower incidence of febrile urinary tract infection as a first symptom of VUR/RN (P<0.05). We conclude that genetic polymorphisms of RAS components are not independent prognostic indicators of renal scarring in patients with VUR. PMID- 12579400 TI - Autosomal recessive polycystic kidney disease: outcomes from a single-center experience. AB - Autosomal recessive polycystic kidney disease (ARPKD) is a relatively common form of pediatric polycystic kidney disease with an incidence of 1:20,000 live births. Previous reports, primarily from populations of European origin, indicate that the clinical presentation and disease course are quite variable. Using a retrospective study design, we sought to determine whether the clinical course and outcome of our multi-ethnic patient cohort differs from the published literature. A 10-year (1990-2000) retrospective study was conducted in which we reviewed the clinical, histopathological, and imaging records of our 31 ARPKD patients. Patients were diagnosed between 0 and 14 years of age, with 17 (55%) presenting within the 1st month of life. The mean follow-up was 67 months and age at last follow-up ranged from 0.5 to 16 years. Of the 17 patients diagnosed as neonates, 11 (65%) had respiratory insufficiency complicated by pneumothoraces. Two died shortly after birth and 2 died within the 1st year of life due to respiratory failure. Among the 13 neonatal survivors, 7 (54%) developed progressive renal insufficiency, whereas 6 of 14 (43%) of those children who presented beyond 1 month of age developed renal insufficiency. Hypertension was present in 55% of our patients, with nearly all neonatal survivors requiring antihypertensive management. Evidence of portal hypertension was found in 10 (37%) of the 27 patients who survived the 1st year of life. In our multi-ethnic ARPKD cohort, the 1-year survival rate (87%) and the clinical variability are comparable to those previously reported. With the recent identification of the PKHD1 gene, characterization of disease-causing mutations should provide new insights into the molecular basis for this phenotypic variability. PMID- 12579401 TI - Erythrocyte deformability and microhematuria in children and adolescents. AB - The aim of this study was to investigate whether erythrocyte deformability is higher in otherwise healthy children and adolescents with asymptomatic isolated glomerular microhematuria and no clinical or laboratory signs indicating renal disorder. Erythrocyte deformability in 33 children and adolescents with unexplained asymptomatic isolated glomerular microhematuria (study group) was compared with erythrocyte deformability in 133 individuals without urinary tract disease and without microhematuria (control group), 26 patients with microhematuria and IgA nephropathy (IgA nephropathy group), and 31 children and adolescents with benign familial hematuria (benign familial hematuria group). The erythrocyte and buffer filtration time was measured, the erythrocyte-to-buffer filtration time ratio calculated, and, on the basis thereof, erythrocyte deformability was estimated as an inverse proportion of the erythrocyte-to-buffer filtration time ratio in all subjects. The erythrocyte-to-buffer filtration time ratio increased with age in all groups (P<0.001). In the study group, the mean erythrocyte-to-buffer filtration time ratio, corrected for age of subjects, was significantly lower than in the control (P=0.042), IgA nephropathy (P=0.004), and benign familial hematuria groups (P= 0.048). Erythrocytes with higher deformability probably pass more easily through the glomerular basement membrane that is assumed to be normal, in which case the mechanism of asymptomatic isolated glomerular microhematuria could be explained by higher erythrocyte deformability. PMID- 12579402 TI - Bone alterations in children with idiopathic hypercalciuria at the time of diagnosis. AB - Some children with idiopathic hypercalciuria (IH) develop bone alterations at some stage of the disease. The aims of this study were to evaluate bone mass in 88 children with IH (G1) at the time of diagnosis and to compare the findings with data for a control group of 29 normal children (G2). Kidney and bone metabolism markers were measured in both groups, and bone densitometry was performed. Serum alkaline phosphatase, intact parathyroid hormone, urinary calcium and uric acid were significantly higher in G1, whereas urinary volume and urinary citrate excretion were lower. The following densitometric parameters were significantly lower in G1: (1) lumbar spine (L(2)-L(4)) bone mineral density (BMD), bone mineral content (BMC), BMC corrected for height and for width of the vertebra, volumetric BMD (BMDvol), and Z score; (2) whole-body BMD; (3) femoral neck BMD. Lumbar spine BMDvol was reduced (osteopenia) in 35% of the patients compared with G2. N telopeptide, a urinary marker of bone resorption, was significantly higher in G1 than in G2, and was negatively correlated with lumbar spine BMD and BMDvol. Children with urinary lithiasis or idiopathic hyperuricosuria associated with IH showed no significant differences in bone metabolism compared with children without these associations. We conclude that (1) there is an altered bone metabolism in IH, with osteopenia already present at diagnosis in 35% of the patients; (2) N telopeptide is one of the most useful markers of bone alterations in IH, especially at an early stage of the disease; (3) investigation of bone metabolism is necessary in IH to prevent future serious consequences such as osteoporosis and bone fractures. PMID- 12579403 TI - Urinary mineral excretion in healthy Iranian children. AB - The purpose of this study was to determine normal reference values for urinary calcium/creatinine (Ca/Cr), phosphate/creatinine (P/Cr), magnesium/creatinine (Mg/Cr), sodium/creatinine (Na/Cr), potassium/creatinine (K/Cr), sodium/potassium (Na/K), calcium/sodium (Ca/Na), and uric acid/creatinine ratios in healthy Iranian children. Nine hundred and ninety children (515 boys, 475 girls) aged 1 month to 14 years were studied. Two non-fasting random urine specimens (1 week apart) from each subject and 24-h urine collections from 114 children were analyzed for Ca, P, Mg, uric acid, Na, K, and Cr. Urinary Ca/Cr, P/Cr, Mg/Cr, Na/Cr, K/Cr, Na/K, Ca/Na, and uric acid/creatinine ratios were determined from each sample. Non-fasting Ca/Cr, P/Cr, Na/Cr, K/Cr, Na/K, Ca/Na, and uric acid/creatinine ratios were not significantly different between the sexes (P>0.05). Urinary Mg/Cr ratios were higher in girls than boys (P<0.001). No significant relationships were found between urinary Ca/Cr and urinary Na/Cr, urinary Ca/Cr and urinary Na/K, and urinary Ca/Cr and urinary Ca/Na (P>0.05). The P/Cr values showed a gradual decrease with age (from mean+/-SD 0.962+/-0.172 mg/mg at 1 month of life to 0.318+/-0.124 mg/mg at 14 years) (P<0.05). The Ca/Cr ratio was highest between 6 months and 3 years (mean+/-SD=0.047+/-0.041 mg/mg). Following a moderate decrease it stabilized by the age of 7 years (mean+/ SD=0.038+/-0.044 mg/mg). Urinary ratios of Mg/Cr were significantly higher in children under 10 years (mean+/-SD=0.042+/-0.015 mg/mg) compared with the 11- to 14-year age group (mean+/-SD=0.031+/-0.001 mg/mg) (P<0.05). The uric acid/creatinine ratios decreased from 0.060+/-0.077 mg/mg in those less than 7 years to 0.041+/-0.033 in the 11- to 14-year group. Urinary Na/Cr ratios were significantly lower in younger age groups compared with the older age group (P<0.05). Urinary K/Cr ratio was highest in younger children, and then steadily decreased with age. There was no correlation between 24-h urinary Ca and Na excretion. The mean Ca/Na ratios significantly decreased with advancing age (P<0.05). The solute/creatinine ratios in the non-fasting urine samples correlated well with the 24-h solute excretion. We provide reference values for urinary Ca/Cr, P/Cr, Mg/Cr, Na/K, K/Cr, and uric acid/creatinine ratios in normal Iranian children. A child's age and ethnicity should be taken into consideration when assessing the urinary solute/creatinine ratios. PMID- 12579404 TI - Office and ambulatory blood pressure elevation in children with chronic renal failure. AB - Hypertension is a frequent complication in patients with chronic renal insufficiency (CRI) and is associated with target organ damage, including left ventricular hypertrophy (LVH). To better assess hypertension in pediatric patients with CRI, we performed 24-h ambulatory blood pressure monitoring (ABPM) and evaluated the relationship between ABPM parameters and LVH in 29 children, mean age 12.4+/-3.8 years. There was no significant difference in the frequency of hypertension comparing casual systolic blood pressure (SBP) (21%) with the mean 24-h (21%) or daytime mean SBP (21%). However, diastolic hypertension was detected more frequently using ABPM: 24% for 24-h diastolic blood pressure (DBP), 14% for daytime DBP, and 7% for casual DBP. Nighttime systolic hypertension as well as diastolic hypertension was detected in 12 (41%) children. Seventeen (59%) patients had attenuated dipping for SBP and 9 (31%) had abnormal DBP dipping. Lower SBP dipping was associated with lower glomerular filtration rate (r=0.44, P<0.05). LVH was found in 6 (21%) patients. LVM index was significantly correlated with 24-h SBP (r=0.43, P<0.05). Multiple regression analysis confirmed that higher 24-h SBP was the only independent predictor for increased LVM index (P=0.001). No significant relationship was found between LVM index and office blood pressure. These results confirm a high prevalence of blood pressure abnormalities using ABPM criteria in children with CRI and suggest that ABP may better predict end-organ damage in these patients than casual BP. PMID- 12579405 TI - Renin-angiotensin gene polymorphism in children with uremia and essential hypertension. AB - The M235T polymorphism of the angiotensinogen (ANG) gene, the I/D polymorphism of the angiotensin converting enzyme (ACE) gene, and the A1166C polymorphism of the angiotensin II type 1 receptor (AT1R) gene were identified in 70 patients with end-stage renal disease [20 pediatric ESRD, aged 14.9+/-3.1, years blood pressure (BP) 139+/-14/91+/-13 mmHg, 50 adult ESRD, aged 48.7+/-18.7 years, BP 149.1+/ 24/96.9+/-12 mmHg], 35 with juvenile essential hypertension (JEHT, aged 14.4+/ 2.7 years, 24-h mean BP 135.37+/-7.37/72.4+/-7.68 mmHg), 130 adult healthy normotensive controls (aged 34.9+/-8.1 years, BP 117.8+/-8.7/78.7+/-8.5 mmHg), and 20 pediatric controls (aged 13.2+/-1.2 years, BP 109+/-6.5/71+/-5.9 mmHg). The ACE gene polymorphism was determined by polymerase chain reaction and the ANG and AT1R gene polymorphisms by single-step LightCycler technology. The ACE gene distribution of the Hungarian controls did not differ from the results of the other Caucasian populations. In JEHT and pediatric ESRD patients, the MT genotype of ANG was more frequent than in controls (JEHT 80%, pediatric ESRD 74% versus controls 50%, P<0.02). The DD genotype of ACE was over-represented in pediatric ESRD compared with controls (ESRD 45% versus controls 22%, P<0.05). There was a non-significant increase in the CC genotype frequency of AT1R in adult patients with ESRD compared with controls. In conclusion, there was an increased frequency of the ACE DD genotype in pediatric ESRD, which could be a genetic risk factor for the development of ESRD. Furthermore, there was a significant increase in MT genotype frequency of ANG M235T polymorphism in pediatric ESRD and JEHT. The role of AT1R gene polymorphism needs further investigation. PMID- 12579406 TI - Hypertension in hypophosphatemic rickets--role of secondary hyperparathyroidism. AB - Hypertension has been anecdotally reported in children with familial hypophosphatemic rickets (XLH). To better identify and characterize the clinical and laboratory features of hypertensive XLH children, we reviewed the medical records of 41 XLH children, all treated with phosphate and vitamin D analogues. Eight children, who were originally normotensive, developed hypertension during the 2nd decade of life. At diagnosis of hypertension all had persistent secondary/tertiary hyperparathyroidism (HPTD), defined as high serum parathyroid hormone (PTH) for 12 months or longer. Seven had nephrocalcinosis (NC). Analysis of data showed that of 11 children with HPTD, 8 developed hypertension compared with 0 among 30 without HPTD (P<0.001). Of 40 children studied, 18 had NC that was significantly associated with both HPTD (P<0.01) and hypertension (P<0.025). At diagnosis of hypertension, serum calcium was elevated in 2. Plasma renin activity was high in 3 of 4 patients in whom it was measured. Doppler ultrasonography or renal scan was normal in the 5 children studied. Early echocardiography showed left ventricular hypertrophy in only 2 of 5 children studied. In 3 patients who underwent parathyroidectomy, hypertension persisted and 1 progressed to renal failure. Serum creatinine remained normal in all others. Successful treatment of hypertension consisted of beta-adrenergic blockers, angiotensin converting enzyme inhibitors, and Ca channel blockers as monotherapy or in combination. We conclude that hypertension in treated XLH children is closely associated with HPTD. Emphasis should therefore be placed on prevention of the development of HPTD as a complication of XLH treatment, and close monitoring for hypertension in those who do develop HPTD. PMID- 12579407 TI - Complications of peritoneal dialysis in children with Eagle-Barrett syndrome. AB - Eagle Barrett syndrome (EBS) is characterized by the triad of abdominal muscle deficiency, urinary tract abnormalities, and cryptorchidism. Approximately 25% of patients with EBS progress to end-stage renal disease. It is speculated that the abdominal muscular defects in EBS pose technical problems in achieving successful peritoneal dialysis (PD). In this retrospective analysis, we reviewed the medical records of EBS and non-EBS PD patients cared for at Rainbow Babies and Children's Hospital from 1985 to 2002; 5 EBS and 9 non-EBS patients were analyzed. PD duration, total complication rates, and catheter usage rates in the two groups were not significantly different. The two most frequent complications were peritonitis and catheter mechanical malfunction during 103 patient-months in EBS patients and 296 patient-months in non-EBS patients. Peritonitis occurred 1 episode every 20.6 patient-months and 14.8 patient-months in EBS and non-EBS patients, respectively. The time from PD initiation to onset of any complication, including first peritonitis, was not significantly different in the two groups. Although the age at PD initiation was significantly different between the groups, there was no correlation between age at onset of PD and complication rates or time to first complication. Despite their abdominal muscle defects, EBS patients do not have more-frequent PD complications. PMID- 12579409 TI - Twenty-one years of developmental nephrology: the kidney then and now. AB - Developmental renal physiology was guided for a long time by the notion that the "immature" kidney is characterized by glomerular-tubular imbalance, with glomerular preponderance. In accordance with this concept, the filtering capacity of the developing kidney exceeds the ability of the renal tubules to handle the filtrate, resulting in the urinary loss of substances such as amino acids and bicarbonate. Estimates of age-related changes in glomerular volume, based on measurements of glomerular diameter, and of proximal tubular volume, based on measurements of tubular length and tubular diameter, appeared to support this contention. The experimental approach that led to these conclusions was based on the assumptions that the increase in glomerular size is distributed evenly between the vascular and nonvascular elements and that tubular volume increases pari passu with the luminal surface area. Both assumptions were proved wrong. Moreover, micropuncture studies performed in guinea pigs and rats revealed that the proportionality between glomerular filtration and proximal reabsorption of fluid (i.e., glomerulo-tubular balance) is maintained throughout development. Subsequent studies showed that several transport mechanisms function quite adequately from the first days of extrauterine life. Sodium is avidly reabsorbed in distal nephron segments, under the stimulus of the high levels of aldosterone present during infancy. The increase in the secretion of potassium that would occur in the adult under these circumstances is mitigated by the low expression of potassium channels. In the case of phosphate, there is enhanced reabsorption, particularly at the level of the proximal tubule, due in part to a growth specific NaPi type II transporter. These different adaptive mechanisms converge towards the maintenance of a positive external balance for substances that are essential to growth. The emergence of cellular and molecular biology has also encouraged research in the field of renal morphology. The genes that control mesenchymal-epithelial interactions and the signaling factors that mediate their effect have been described. Progress is being made in the identification of genes involved in certain forms of renal malformations. Novel methods of investigation, such as DNA arrays, are likely to lead to an even fuller, dynamic portrayal of gene expression during nephrogenesis. Characterizing the functional correlates of these genes will require investigators who are not only proficient in molecular biology, but who are also masters of physiological methods. PMID- 12579410 TI - Proceedings of the Eighth International Workshop on Developmental Nephrology: genes, morphogenesis, and function. The sessions. PMID- 12579411 TI - Payment for organ donation: unacceptable or a possible solution? PMID- 12579412 TI - Acute renal failure in Kawasaki disease. PMID- 12579416 TI - Skin pigmentation, biogeographical ancestry and admixture mapping. AB - Ancestry informative markers (AIMs) are genetic loci showing alleles with large frequency differences between populations. AIMs can be used to estimate biogeographical ancestry at the level of the population, subgroup (e.g. cases and controls) and individual. Ancestry estimates at both the subgroup and individual level can be directly instructive regarding the genetics of the phenotypes that differ qualitatively or in frequency between populations. These estimates can provide a compelling foundation for the use of admixture mapping (AM) methods to identify the genes underlying these traits. We present details of a panel of 34 AIMs and demonstrate how such studies can proceed, by using skin pigmentation as a model phenotype. We have genotyped these markers in two population samples with primarily African ancestry, viz. African Americans from Washington D.C. and an African Caribbean sample from Britain, and in a sample of European Americans from Pennsylvania. In the two African population samples, we observed significant correlations between estimates of individual ancestry and skin pigmentation as measured by reflectometry (R(2)=0.21, P<0.0001 for the African-American sample and R(2)=0.16, P<0.0001 for the British African-Caribbean sample). These correlations confirm the validity of the ancestry estimates and also indicate the high level of population structure related to admixture, a level that characterizes these populations and that is detectable by using other tests to identify genetic structure. We have also applied two methods of admixture mapping to test for the effects of three candidate genes (TYR, OCA2, MC1R) on pigmentation. We show that TYR and OCA2 have measurable effects on skin pigmentation differences between the west African and west European parental populations. This work indicates that it is possible to estimate the individual ancestry of a person based on DNA analysis with a reasonable number of well defined genetic markers. The implications and applications of ancestry estimates in biomedical research are discussed. PMID- 12579417 TI - An Alu-mediated rearrangement as cause of exon skipping in Hunter disease. AB - Hunter syndrome (Mucopolysaccharidosis type II), a rare X-linked lysosomal storage disorder, results from deleterious mutations in the iduronate-2-sulfatase ( IDS) gene located on Xq27.3-q28. Partial or complete deletions and large rearrangements have been extensively reported in the IDS gene as the basis of Hunter disease. The present report, however, is the first report on a Hunter patient in which Alu-mediated recombinations are implicated. Our patient showed the skipping of exon 8 at the cDNA level, without any splice-junction defects at the genomic level, where a new large rearrangement was identified instead. This new mutant allele consisted of an extensive deletion of IDS sequence of about 3 kb, as well as an additional inserted sequence of 157 bp. Two different computer programs were necessary to elucidate the nature of the insert. NCBI-BLAST query detected a single match for 126 bp out of 157 of the fragment that aligned exactly with a specific chromosomal region, Xq25-27.1, where an AluSg sequence is adjacent to an L1. Instead, the Repeat Masker program identified only 83 bp out of 157 of the insert, which was confirmed as an AluS. The observed homology between the AluSc sequence in the IDS intron 8 and the inserted AluS element, as well as the closeness of 26 bp Alu core sequence, considered to be a recombination hotspot, made us hypothesise upon the fact that both an Alu retrotransposition and an Alu-mediated deletion underlie the disease-producing rearrangement. We, therefore, now propose a mechanism that led to the large genomic deletion causing the production of the aberrant mRNA splicing. PMID- 12579419 TI - Obtaining consent for research studies on incompetent subjects: the Quebec experience. PMID- 12579420 TI - Influence of tidal volume on left ventricular stroke volume variation measured by pulse contour analysis in mechanically ventilated patients. AB - OBJECTIVE: Real-time measurement of stroke volume variation by arterial pulse contour analysis (SVV) is useful in predicting volume responsiveness and monitoring volume therapy in mechanically ventilated patients. This study investigated the influence of the depth of tidal volume (V(t)) on SVV both during the state of fluid responsiveness and after fluid loading in mechanically ventilated patients. DESIGN AND SETTING: Prospective study in a university hospital, adult cardiac surgery intensive care unit. PATIENTS AND PARTICIPANTS: 20 hemodynamically stable patients immediately after cardiac surgery. INTERVENTIONS: Stepwise fluid loading using colloids until stroke volume index (SVI) did not increase by more than 10%. Before and after fluid loading V(t) was varied (5, 10, and 15 ml/kg body weight) in random order. MEASUREMENTS AND RESULTS: Pulse contour SVV was measured before and after volume loading at the respective V(t) values. Thirteen patients responded to fluid loading with an increase in SVI greater than 10%, which confirmed volume responsiveness at baseline measurements. These were included in further analysis. During volume responsiveness SVV at V(t) of 5 ml/kg (7+/-0.7%) and SVV at V(t) of 15 ml/kg (21+/-2.5%) differed significantly from that at V(t) of 10 ml/kg (15+/-2.1%). SVV was correlated significantly with the magnitude of V(t). After volume resuscitation SVV at the respective V(t) was significantly reduced; further, SVV at V(t) of 5 ml/kg(-1) (5.3+/-0.6%) and 15 ml/kg (16.2+/-2.0%) differed significantly from that at V(t) of 10 ml/kg (10.2+/-1.0%). SVV and depth of V(t) were significantly related. CONCLUSIONS: In addition to intravascular volume status SVV is affected by the depth of tidal volume under mechanical ventilation. This influence must be regarded when using SVV for functional preload monitoring. PMID- 12579421 TI - Fatal air embolism in an extremely low birth weight infant: can it be caused by intravenous injections during resuscitation? AB - OBJECTIVE: A preterm infant with a birth weight of 384 g who succumbed to vigorous resuscitation for sudden respiratory failure on the third day of life is presented. Postmortem examination revealed, apart from extensive pulmonary hemorrhage, a fatal amount of air mainly in the right ventricle. We believe that this air had been introduced via peripheral venous cannulas due to inadvertent and unavoidable air admixture to each injection volume administered with a syringe, and we develop an experimental model to confirm that fatal amounts of air can indeed accumulate with frequent change over of syringes within a short period of time. DESIGN: An empty 50-ml syringe was connected to a 15-cm-long connection line via a three-way tap. With a 1 ml syringe 100 doses of 0.5 ml aqua were injected into the connection line. The amount of air which had collected in the 50 ml syringe after the 100 injection cycles was measured. This process was repeated three times each by three of the authors and the average air volume introduced with 100 injections calculated for each investigator. RESULTS: The average amounts of air which had entered the closed system after 100 acts of syringe assembly and aqua administration were 1.84, 1.95, and 2.0 ml. This corresponds to an average volume of almost 0.02 ml per injection. PMID- 12579422 TI - Mapping of a gene responsible for the difference in amylopectin structure between japonica-type and indica-type rice varieties. AB - The present investigation revealed that the alk and gel(t) genes, which cause the differences between a japonica rice variety Nipponbare and an indica rice variety Kasalath in terms of the disintegration of endosperm starch granules in alkali solution and their gelatinisation in a 4 M urea solution, respectively, cosegregated in backcross inbred lines derived from a cross between the two varieties. The segregation pattern of the profile for amylopectin chain-length, which was distinguished by enrichment in short chains of DP<==11 and depletion in intermediate-size chains of 12<==DP<==24 in japonica as compared with indica, was exactly the same as those of the above physico-chemical properties of starch granules, and the gene was designated as acl(t). Gene-mapping analysis showed that the starch synthase IIa ( SSIIa) gene is located at the alk locus on chromosome 6 in the rice genome. These results lead us to the possibility that different alleles of the SSIIa gene are responsible for differences in amylopectin structure between the two varieties, in that SSIIa plays a distinct role in the elongation of short chains within clusters (A+B(1) chains) of amylopectin. It is proposed that the activity of SSIIa in japonica rice is reduced in amount or functional capacity relative to the activity of this enzyme in indica rice. This, in turn, would explain why starch from japonica rice has a lower gelatinisation temperature than starch from indica rice and is more susceptible to disintegration in alkali or urea. The evidence for this hypothesis is that the alk(t), gel(t), acl(t) and SSIIa genes all map to the same locus. PMID- 12579423 TI - DNA transfer by highly asymmetric somatic hybridisation in Medicago truncatula (+) Medicago rugosaand Medicago truncatula(+) Medicago scutellata. AB - A regenerable line of Medicago truncatula(Jemalong 2HA) as a recipient species, was fused with the sexually incompatible species Medicago scutellataor Medicago rugosa.The treatments described maintain the chromosome number of the recipient but enable the transfer of small amounts of DNA of the donor species, probably by intergenomic recombination. Without a chromosome number-change fusion products can readily regenerate to produce fertile plants; and potentially a library with a diverse array of new genetic material. The selection of fused cells is based on treatment of the recipient cells with iodoacetamide (IOA), a non-regenerable donor, gamma-irradiation of the donor, and regeneration on a medium favouring the recipient. DNA transfer was demonstrated by amplified fragment length polymorphism (AFLP), Southern hybridisation and changed morphology. PMID- 12579424 TI - Microsatellite polymorphism in natural populations of wild emmer wheat, Triticum dicoccoides, in Israel. AB - Diversity in 20 microsatellite loci of wild emmer wheat, Triticum dicoccoides, was examined in 15 populations (135 genotypes) representing a wide range of ecological conditions of soil, temperature, and water availability, in Israel and Turkey. An extensive amount of diversity at microsatellite loci was observed despite the predominantly selfing nature of this plant species. The 20 Gatersleben wheat microsatellites (GWM), representing 13 chromosomes of genomes A and B of wheat, revealed a total of 364 alleles, with an average of 18 alleles per GWM marker (range: 5-26). The proportion of polymorphic loci per population averaged 0.90 (range: 0.45- 1.00); genic diversity, He, averaged 0.50 (range 0.094- 0.736); and Shannon's information index averaged 0.84 (range 0.166-1.307). The coefficients of genetic distance between populations were high and averaged D=1.862 (range 0.876-3.320), an indication of sharp genetic divergence over short distances. Interpopulation genetic distances showed no association with geographic distance between the population sites of origin, which ruled out a simple isolation by distance model. Genetic dissimilarity values between genotypes were used to produce a dendrogram of the relationships among wild wheat populations by the unweighted pair-group method with arithmetic averages (UPGMA). The results showed that all the wild emmer wheat populations could be distinguished. Microsatellite analysis was found to be highly effective in distinguishing genotypes of T. dicoccoides, originating from diverse ecogeographical sites in Israel and Turkey, with 88% of the 135 genotypes correctly classified into sites of origin by discriminant analysis. Our present microsatellite results are non-random and in agreement with the previously obtained allozyme and RAPD patterns, although the genetic-diversity values obtained with microsatellites are much higher. Significant correlates of microsatellite markers with various climatic and soil factors suggest that, as in allozymes and RAPDs, natural selection causes adaptive microsatellite ecogeographical differentiation, not only in coding, but most importantly in non coding genomic regions. Hence, the concept of "junk DNA" needs to be replaced by at least partly regulatory DNA. The obtained results suggest that microsatellite markers are useful for the estimation of genetic diversity in natural populations of T. dicoccoidesand for the tagging of agronomically important traits derived from wild emmer wheat. PMID- 12579425 TI - Analysis of genetic similarity detected by AFLP and coefficient of parentage among genotypes of sugar cane ( Saccharum spp.). AB - Despite the economical importance of sugar cane, until the present-date no studies have been carried out to determine the correlation of the molecular-based genetic similarity (GS) and the coefficient of parentage ( f)-estimates generated for cultivars. A comprehensive knowledge of the amount of genetic diversity in parental cultivars, could improve the effectiveness of breeding programmes. In this study, amplified fragment length polymorphism (AFLP) and pedigree data were used to investigate the genetic relationship in a group of 79 cultivars (interspecific hybrids), used as parents in one of the Brazilian breeding programmes, and four species of Saccharum ( Saccharum sinense, Saccharum barberi and two of Saccharum officinarum). The objectives of this study were to assess the level of genetic similarity among the sugar-cane cultivars and to investigate the correlation between the AFLP-based GS and f, based on pedigree information. Twenty one primer combinations were used to obtain the AFLP molecular markers, generating a total of 2,331 bands, of which 1,121 were polymorphic, with a polymorphism rate, on average, of 50% per primer combination. GSs were determined using Jaccard's similarity coefficient, and a final dendrogram was constructed using an unweighted pair-group method using arithmetic average (UPGMA). AFLP based GS ranged from 0.28 to 0.89, with a mean of 0.47, whereas f ranged from 0 to 0.503, with a mean of 0.057. Cluster analysis using GS divided the genotypes into related subgroups suggesting that there is important genetic relationship among the cultivars. AFLP-based GS and f were significantly correlated ( r= 0.42, P< 0.001), thus the significance of this r value suggests that the AFLP data may help to more-accurately quantify the degree of relationship among sugar-cane cultivars. PMID- 12579426 TI - Study of the relationship between pre-harvest sprouting and grain color by quantitative trait loci analysis in a whitexred grain bread-wheat cross. AB - In many wheat ( Triticum aestivumL.) growing areas, pre-harvest sprouting (PHS) may cause important damage, and in particular, it has deleterious effects on bread-making quality. The relationship between PHS and grain color is well known and could be due either to the pleiotropic effect of genes controlling red-testa pigmentation ( R) or to linkage between these genes and other genes affecting PHS. In the present work, we have studied a population of 194 recombinant inbred lines from the cross between two cultivars, 'Renan' and 'Recital', in order to detect QTLs for both PHS resistance and grain color. The variety 'Renan' has red kernels and is resistant to PHS, while 'Recital' has white grain and is highly susceptible to PHS. A molecular-marker linkage map of this cross was constructed using SSRs, RFLPs and AFLPs. The population was evaluated over 2 years at Clermont-Ferrand (France). PHS was evaluated on mature spikes under controlled conditions and red-grain color was measured using a chromameter. Over the 2 years, we detected four QTLs for PHS, all of them being co-localized with QTLs for grain color. Three of them were located on the long arm of chromosomes 3 A, 3B and 3D, close to the loci where the genes R and taVp1 were previously mapped. For these three QTLs, the resistance to PHS is due to the allele of the variety 'Renan'. Another co-located QTL for PHS and grain color was detected on the short arm of chromosome 5 A. The resistance for PHS for this QTL is due to the allele of 'Recital'. PMID- 12579427 TI - Evidence for a prokaryotic insertion-sequence contamination in eukaryotic sequences registered in different databases. AB - An insertion-sequence of prokaryotic origin was detected in a genomic clone obtained from a Phaseolus vulgaris bacterial artificial chromosome (BAC) library. This BAC clone, characterized as part of a contig constructed near a virus resistance gene, exhibited restriction fragment length polymorphism with an overlapping clone of the contig. Restriction analysis of DNA obtained from individual colonies of the stock culture indicated the presence of a mixed population of wild-type and insertional mutants. Sequence analysis of both members of the population revealed the presence of IS 10R, an insertion-sequence from Escherichia coli. A BLAST search for IS 10-like sequences detected unexpected homologies with a large number of eukaryotic sequences from Homo sapiens, Arabidopsis thaliana, Drosophila melanogasterand Caenorhabditis elegans. Southern analysis of a random sample of BAC clones failed to detect IS 10 in the BAC DNA. However, prolonged sub-culturing of a set of 15 clones resulted in transposition into the BAC DNA. Eventually, all cultures acquired a 2.3-kb fragment that hybridized strongly with IS 10. Sequence analysis revealed the presence of a preferred site for transposition in the BAC vector. These results indicate that a large number, if not all, of the BAC libraries from different organisms are contaminated with IS 10R. The source of this element has been identified as the DH10B strain of E. coli used as the host for BAC libraries. PMID- 12579428 TI - A DNA marker closely linked to the vrs1 locus (row-type gene) indicates multiple origins of six-rowed cultivated barley ( Hordeum vulgare L.). AB - The origin of six-rowed cultivated barley was studied using a DNA marker cMWG699 closely linked to the vrs1 locus. Restriction patterns of the PCR-amplified product of the cMWG699 locus were examined in 280 cultivated ( Hordeum vulgare ssp. vulgare) and 183 wild ( H. vulgare ssp. spontaneum) barleys. Nucleotide sequences of the PCR products were also examined in selected accessions. Six rowed cultivated barleys were divided into two distinct groups, types I and II. Type I six-rowed cultivated barley was distributed widely while type II six-rowed cultivated barley was found only in the Mediterranean region. The type I sequence was also found in a wild barley accession from Turkmenistan whereas the type II sequence was also found in a two-rowed cultivated barley from North Africa and a wild barley from Morocco. These results suggested that the six-rowed type I and II barleys were derived from two-rowed type I and II barleys, respectively, by independent mutations at the vrs1 locus. PMID- 12579429 TI - Evaluating the potential of SSR flanking regions for examining taxonomic relationships in the Vitaceae. AB - Three EST-derived microsatellite loci from Vitis vinifera were amplified and sequenced across eight species of Vitaceae from four different genera. Phylogenetic analysis of the microsatellite's flanking regions produced informative results in congruence with previous studies. Generic relationships were respected and the data produced sufficient inter-specific variation to distinguish between Cayratia acris and Cayratia saponaria, two very closely related species. Overall, the sequence alignments showed that priming sites were conserved, whereas microsatellite repeats were present in most cases but structurally variable. The sequence data provided information on the evolutionary patterns of various microsatellite repeats and their correlation to evolutionary relationships among taxa. PMID- 12579430 TI - Quantitative trait loci associated with traits determining grain and stover yield in pearl millet under terminal drought-stress conditions. AB - Drought stress during the reproductive stage is one of the most important environmental factors reducing the grain yield and yield stability of pearl millet. A QTL mapping approach has been used in this study to understand the genetic and physiological basis of drought tolerance in pearl millet and to provide a more-targeted approach to improving the drought tolerance and yield of this crop in water-limited environments. The aim was to identify specific genomic regions associated with the enhanced tolerance of pearl millet to drought stress during the flowering and grain-filling stages. Testcrosses of a set of mapping population progenies, derived from a cross of two inbred pollinators that differed in their response to drought, were evaluated in a range of managed terminal drought-stress environments. A number of genomic regions were associated with drought tolerance in terms of both grain yield and its components. For example, a QTL associated with grain yield per se and for the drought tolerance of grain yield mapped on linkage group 2 and explained up to 23% of the phenotypic variation. Some of these QTLs were common across stress environments whereas others were specific to only a particular stress environment. All the QTLs that contributed to increased drought tolerance did so either through better than average maintenance (compared to non-stress environments) of harvest index, or harvest index and biomass productivity. It is concluded that there is considerable potential for marker-assisted backcross transfer of selected QTLs to the elite parent of the mapping population and for their general use in the improvement of pearl millet productivity in water-limited environments. PMID- 12579432 TI - The development of a genetic map for meadowfoam comprised of amplified fragment length polymorphisms. AB - Limnanthes alba Benth. (meadowfoam), a diploid ( x=5) winter annual, produces novel very long-chain seed oils (C(20) and C(22)) with less than 2% saturated fatty acids. The first genetic map of meadowfoam, a recently domesticated species, is described herein. Two phenotypically diverse inbred lines, OMF40-11 ( L. alba ssp. alba) and OMF64 ( L. alba ssp. versicolor), were screened for amplified fragment length polymorphisms (AFLPs) using 16 primer combinations. Twenty three percent of the AFLP bands (415 out of 1,801) were polymorphic between OMF40-11 and OMF64. One hundred (OMF40-11xOMF64)xOMF64 BC(1) progeny were genotyped for 107 polymorphic AFLP markers produced by nine AFLP primer combinations. One hundred and three AFLP loci amalgamated into five linkage groups with 14 to 28 loci per linkage group (four loci segregated independently). The map was 698.5-cM long with a mean interlocus spacing of 6.7 cM and no dense clustering of loci. The segregation ratios for 25 loci (23.2%) were significantly distorted. Twenty one of the distorted loci (84%) had an excess of L. alba ssp. versicolor (recurrent parent) alleles. The distorted loci, apart from one locus on linkage group 4, were distally clustered on both ends of linkage groups 1, 4 and 5. The development of the map was facilitated by the small chromosome number, an abundance of restriction site polymorphisms between the two subspecies (23%), and a high multiplex ratio of the AFLP markers (112 per primer combination). PMID- 12579431 TI - Molecular mapping of QTLs for Fusarium head blight resistance in spring wheat. I. Resistance to fungal spread (Type II resistance). AB - Fusarium head blight (FHB, scab) is a fungal disease of wheat and other small cereals that is found in both temperate and semi-tropical regions. FHB causes severe yield and quality losses, but the most-serious concern is the possible mycotoxin contamination of cereal food and feed. Breeding for FHB resistance by conventional selection is feasible, but tedious and expensive. This study was conducted to identify and map DNA markers associated with FHB resistance genes in wheat. A population of 364 F(1)-derived doubled-haploid (DH) lines from the cross 'CM-82036' (resistant)/'Remus' (susceptible) was evaluated for Type II resistance (spread within the spike) during 2 years under field conditions. Marker analysis was performed on 239 randomly chosen DH lines. Different marker types were applied, with an emphasis on AFLP and SSR markers. Analysis of variance, as well as simple and composite interval mapping, were applied. Three genomic regions were found significantly associated with FHB resistance. The most-prominent effect was detected on the short arm of chromosome 3B, explaining up to 60% of the phenotypic variance for Type II FHB resistance. A further QTL was located on chromosome 5A and a third one on 1B. The QTL regions on 3B and 5A were tagged with flanking SSR markers, the 1B QTL was found associated with the high molecular-weight glutenin locus. These results indicate that FHB resistance is under control of a few major QTLs operating together with unknown numbers of minor genes. Marker-assisted selection for these major QTLs involved in FHB resistance appears feasible and should accelerate the development of resistant and agronomically improved wheat cultivars. PMID- 12579433 TI - EST analysis in barley defines a unigene set comprising 4,000 genes. AB - We report the generation of 13,109 EST (Expressed Sequence Tag) sequences from barley as a first step towards the generation of a unigene set for this organism. Sequences were generated from three libraries encompassing 7,568 cDNA clones. Comparisons to nucleic acid and protein sequence databases enabled the assignment of putative functions to the mRNAs. The results of the searches against protein databases were parsed and built into a regularly updated database, available over the World Wide Web. The Stack_Pack clustering system has been applied to survey the level of redundancy, which was calculated to amount to 69%, thus we identified 4,000 different barley genes. To prove the usability of the results of the clustering process for further experiments, we subjected alignments with sequences similar to elongation factor 1 alpha to additional analysis. These sequences represented the largest group with identical putative functions (228 members) and clustering based on the analysis of 3; sequences subdivided the group into five different assemblies. Alignments of the consensus sequences facilitated the development of PCR assays suitable for genetic mapping of four of the different gene-family members, which reside on chromosomes 2H, 4H and 5H, thus demonstrating the suitability of the cluster-results as a basis for in-depth analyses of barley gene families. PMID- 12579434 TI - Evaluation of Hbr (MITE) markers for assessment of genetic relationships among maize ( Zea mays L.) inbred lines. AB - Recently, a new type of molecular marker has been developed that is based on the presence or absence of the miniature inverted repeat transposable element (MITE) family Heartbreaker ( Hbr) in the maize genome. These so-called Hbr markers have been shown to be stable, highly polymorphic, easily mapped, and evenly distributed throughout the maize genome. In this work, we used Hbr-derived markers for genetic characterization of a set of maize inbred lines belonging to Stiff Stalk (SS) and Non-Stiff Stalk (NSS) heterotic groups. In total, 111 markers were evaluated across 62 SS and NSS lines. Seventy six markers (68%) were shared between the two groups, and 25 of the common markers occurred at fairly low frequency ( 4 Tesla). FMRI-studies in healthy human volunteers and psychiatric patients focus on cerebral activity following acute drug administration (single challenge) and on adaptive effects of the CNS due to long- term medication. Their results provide insights into brain physiology and neuropharmacological mechanisms which are in turn relevant for preclinical pharmacological studies, responder analyses and for the investigation of pathogenetic models in psychiatric diseases. However, with these new opportunities, additional methodological considerations and limitations emerge. Besides the need of controlling motion artefacts, the influence of interfering psychological variables, an exact specification of the experimental design, a standardised analysis for data adjustment and technical limitations have to be considered. This article provides an overview of the underlying model of brain function, present applications, future possibilities and methodological limitations of fMRI for the understanding of human psychopharmacology. PMID- 12579472 TI - [Dynamic short-term psychotherapy between support and interpretation]. AB - Interpersonal approaches based on the classic concepts of psychoanalysis draw their significance not only from guiding therapists but also provide instruments for empirical psychotherapy research, particularly focussing on the therapeutic process itself. The Dusseldorf Short Term Psychotherapy Project uses several strategies of the SASB-CMP method to examine the interrelation among certain groups of disorders, different courses of treatment, and outcome in dynamic short term psychotherapy. In addition, this paper uses the concept of CMP-based therapeutic interventions to analyze focus-adherence and offers a system for the classification of other, not focus-related therapeutic interventions. For understanding the effects of dynamic short term psychotherapy, it is essential to consider the distinction between more specific approaches, e. g. interpretation, transference, and insight on the one hand, and more general concepts, e. g. emotional experience, absence of re-traumatization, direct or indirect support on the other hand. The 'principle of interpretation' needs a relatively structured patient's ego, while in other cases the basic (re-)organizing of the patient has to be the crucial goal of therapy. This leads to the assumption that successful outcome is achieved by different ways depending on the patients' differing structural levels. The 2 pilot cases of this study illustrate that based on a higher level of structural functioning, more CMP-specific interpretations in the here and now are used to obtain good results, whereas in a case of lower level the first aim is to strengthen the patient's ego functions and to scrutinize his CMP in the there and then. PMID- 12579473 TI - [Koro--discussing a transcultural psychopathological phenomenon]. AB - Koro describes a psychopathological phenomenon mainly found in Asia. It is the syndrome of an assumption of male genital shrinking. This idea is rarely described in Western culture. Inspired by a current case report--a patient, suffering from schizophrenia described Koro-like symptoms--we will discuss Koro on the basis of a literature review in its transcultural psychiatric context. PMID- 12579476 TI - AlloDerm tympanoplasty of tympanic membrane perforations. AB - PURPOSE: To study the effectiveness of AlloDerm (LifeCell Corporation, Branchburg, NJ) as a graft material in underlay tympanoplasty by comparison to autologous fascia in a chronic tympanic membrane perforation animal model. MATERIALS AND METHODS: Seventeen chinchillas underwent creation of bilateral chronic tympanic membrane perforations over a 6-week period. Twenty-two stable perforations were divided equally between the experimental AlloDerm and control fascia graft groups. The grafts were surgically placed through a postauricular tympanomeatal flap. The tympanic membranes were examined at 4 and 10 weeks and then harvested for histopathological analysis. Tympanoplasty operative times, perforation closure rates, and gross and histological analyses were compared between the AlloDerm and fascia grafts. RESULTS: A statistically significant difference in mean surgical time was recorded between the AlloDerm (47 minutes) and fascia (68 minutes) grafting procedures (t test, P =.001). Perforation closure was achieved in 90% of the AlloDerm and 100% of the fascia treated tympanic membranes. Gross and histopathologic inspections revealed no significant differences. Microscopically, AlloDerm and fascia grafts had similar inflammatory responses, but AlloDerm showed increased fibroblast infiltration and neovascularization. CONCLUSION: The avoidance of donor site morbidity, reduction of surgical time, and excellent gross and histologic outcomes in this animal model reveal that AlloDerm could be a safe, cost-effective alternative to autologous fascia. Further study would be necessary in human clinical trials. PMID- 12579474 TI - Six novel missense mutations in the LDL receptor-related protein 5 (LRP5) gene in different conditions with an increased bone density. AB - Bone is a dynamic tissue that is subject to the balanced processes of bone formation and bone resorption. Imbalance can give rise to skeletal pathologies with increased bone density. In recent years, several genes underlying such sclerosing bone disorders have been identified. The LDL receptor-related protein 5 (LRP5) gene has been shown to be involved in both osteoporosis-pseudoglioma syndrome and the high-bone-mass phenotype and turned out to be an important regulator of peak bone mass in vertebrates. We performed mutation analysis of the LRP5 gene in 10 families or isolated patients with different conditions with an increased bone density, including endosteal hyperostosis, Van Buchem disease, autosomal dominant osteosclerosis, and osteopetrosis type I. Direct sequencing of the LRP5 gene revealed 19 sequence variants. Thirteen of these were confirmed as polymorphisms, but six novel missense mutations (D111Y, G171R, A214T, A214V, A242T, and T253I) are most likely disease causing. Like the previously reported mutation (G171V) that causes the high-bone-mass phenotype, all mutations are located in the aminoterminal part of the gene, before the first epidermal growth factor-like domain. These results indicate that, despite the different diagnoses that can be made, conditions with an increased bone density affecting mainly the cortices of the long bones and the skull are often caused by mutations in the LRP5 gene. Functional analysis of the effects of the various mutations will be of interest, to evaluate whether all the mutations give rise to the same pathogenic mechanism. PMID- 12579475 TI - Results of phase I-II trial of concomitant hyperfractionated radiation and oral etoposide (VP-16) in patients with unresectable squamous cell carcinoma of the head and neck. AB - PURPOSE: The purpose of the current study was to investigate the efficacy of concomitant oral etoposide and hyperfractionated radiation for patients with unresectable head and neck squamous cell carcinoma. METHODS: A prospective nonrandomized phase I-II study was conducted using concomitant oral etoposide (50 mg/d for 13-27 days) and hyperfractionated radiotherapy (1.2 Gy twice daily to a total of 7440 rads) for patients with unresectable squamous cell carcinoma of the head and neck. Toxicity was graded according to the NCI common toxicity criteria. Patients were followed for a minimal period of 2 years. Endpoints for follow-up were recurrence or death. RESULTS: Seventeen patients were enrolled in the study. Grade III hematological toxicity occurred in 1 patient and moderate to severe mucositis occurred in all but 2 patients requiring a gastrostomy tube (n = 13) or intravenous fluids hydration (n = 2). One patient died of cardiac arrest unrelated to the treatment regimen. The overall response rates in patients that finished the protocol were 80% for the primary site and 100% for the neck. A complete response was observed in 47% at the primary site and 33% in the neck. Local control and disease-free survival (DFS) at an average follow-up of 3.7 years were 47% and 29%, respectively. CONCLUSIONS: Concomitant etoposide and hyperfractionated radiation is well tolerated and seems to be effective in the treatment of unresectable HNSCC with acceptable mucosal toxicity. PMID- 12579478 TI - Superiority of bilateral cochlear implantation over unilateral cochlear implantation in tone discrimination in chinese patients. AB - PURPOSE: The benefits of bilateral cochlear implants (CIs) versus unilateral CIs were evaluated by comparing the Cantonese lexical tone discrimination scores conducted in a quiet environment and against a background noise. MATERIALS AND METHODS: Four postlingually deafened Cantonese-speaking adults (2 men and 2 women) with bilateral CIs were included in this study. The subjects were their own control in the monaural hearing condition. Both the Cantonese lexical tone stimuli and the speech-weighted background noise were presented at 0(0) azimuth and at a distance of 1 m from the subject. The speech stimuli, which were maintained at 65 dB sound pressure level, were presented in both a quiet environment and against a background noise at signal to noise ratios (SNRs) of +15, +10, +5, 0, -5, -10, and -15. RESULTS: Against a background noise, the bilateral CIs required +5 dB SNR only to obtain significant scores in discriminating Cantonese lexical tones and to achieve discrimination scores that were comparable to the optimal discrimination scores obtained in quiet. No significant difference in the discrimination scores was observed between binaural and monaural hearing conditions when the tests were conducted in quiet. CONCLUSIONS: Our study showed that in the presence of background noise, bilateral CIs were better than unilateral CIs in discriminating Cantonese lexical tones. PMID- 12579477 TI - Mastoid obliteration by BMP-2/collagen composites: an experimental study using tissue engineering. AB - PURPOSE: Several materials have been used in the application of mastoid cavity obliteration during surgery for cholesteatoma; however, nothing has won universal acceptance. Through the advancement of tissue engineering, bone morphogenetic protein-2 (BMP-2)/collagen composites have been elucidated as inducers of heterogenic bone formation. This study was performed to investigate whether these composites are potentially obliteration materials for use in the mastoid cavity by using an animal experimental study. MATERIALS AND METHODS: The composites were implanted in the rat mastoid to investigate whether new bone would be tissue engineered in the mastoid and, if so, whether the newly formed bone was stable. The composites were examined histologically over a 24-week period. RESULTS: The composites implanted in the rat mastoid were able to tissue engineer new bone, and the newly formed bone was stable as assessed histologically, with almost normal bone structure, that was not resorbed during the 24-week period. Adverse immunological reactions were not found during our observation. CONCLUSIONS: Bone that was tissue engineered by the BMP-2/collagen composites was stable as assessed by histological examination and persisted in the rat mastoid. The present study shows that the composites have the potential to become real materials for use in mastoid obliteration. PMID- 12579479 TI - Second head and neck cancers and tobacco usage. AB - OBJECTIVES: To examine the relative incidence of second primary carcinomas in patients who continued smoking compared with those who had ceased smoking. METHODS: This is a retrospective study based on review of the Wake Forest University-Baptist Medical Center Tumor Registry for the years 1985 through 2000. Ninety-one patients who had had an index head and neck tumor and who developed a second independent head and neck primary tumor, were identified. These cases were grouped into synchronous (different sites within 6 months) and metachronous (different site after 6 months or same site after 3 years) second tumors and were examined with respect to smoking history-specifically whether smoking had continued or ceased after the diagnosis of the index tumor. RESULTS: Of the 91 patients identified with double head and neck tumors, 88 were tobacco users. Comprising the group of 54 patients with metachronous second primaries were 51 smokers-25 who had continued and 26 who had ceased tobacco use. Of the 26 patients who had quit smoking but had developed a second primary, 13 had stopped smoking even before the index primary had been diagnosed. The remaining 13 had stopped when the index primary was treated. CONCLUSIONS: A review of 91 patients with double head and neck primary tumors indicate no difference in the frequency of second tumors developing in a group of patients who continued to smoke after diagnosis of their index cancers relative to patients who stopped smoking. This finding suggests a critical cellular level of cumulative and persistent damage. Methods to reverse this genetic alteration are hypothesized to be potentially more significant than smoking cessation efforts in preventing subsequent head and neck cancers. PMID- 12579481 TI - Endoscopically placed expandable metal tracheal stents for the management of complicated tracheal stenosis. AB - BACKGROUND: Metal stents have been advocated to manage complicated tracheal stenosis. OBJECTIVE: The purpose of this investigation is to review the effectiveness of endoscopic placement of tracheal expandable metal stents for complicated tracheal stenosis. METHODS: The charts of 6 patients who have undergone placement of metal expandable stents between 1998 and 2000 were reviewed. RESULTS: Initially, all patients enjoyed immediate palliation of symptomatic tracheal stenosis. Eventually, 4 patients developed significant granulation tissue and/or recurrent stenosis, requiring intervention within 6 months after placement of the stent. One patient required the removal of the stent and placement of a T-tube silicone stent. CONCLUSIONS: Metal stents provide temporary palliation for tracheal stenosis. Metal stents, however, are associated with a high incidence of obstruction with granulation tissue. Their use should be limited to a select group of patients with a short life expectancy (because of other comorbidities) or patients who are not good candidates for reconstructive surgery and/or who refuse or cannot tolerate a tracheotomy. PMID- 12579480 TI - Laser surgery for the treatment of glottic carcinomas. AB - PURPOSE: The standard treatment for patients with early glottic carcinoma in Israel has been radiotherapy. In recent years, encouraging results with laryngo microscopic carbon dioxide laser surgery as a treatment for early glottic carcinoma has changed our treatment strategy. We conducted a retrospective study to investigate the results of carbon dioxide laser excisional technique for early glottic carcinoma (T1, T2). MATERIALS AND METHODS: Twenty-six had squamous cell carcinoma (SCC), (21 patients with T1 and 5 patients with T2 lesions), 3 had carcinoma in situ, (CIS) and 3 had verrucous carcinoma (VC). RESULTS: All patients were free of disease after salvage treatment at the most recent follow up. CONCLUSIONS: Careful patient selection with endoscopic staging and strict follow-up are essential to secure good results in the treatment of carbon dioxide laser for early laryngeal carcinoma. PMID- 12579482 TI - Postoperative radiation therapy for squamous cell carcinoma of the head and neck. AB - PURPOSE: To discuss the role of postoperative radiation therapy (RT) for patients with squamous cell carcinoma of the head and neck. RESULTS: Patients with adverse pathologic features have a high likelihood of local-regional recurrence and a decreased probability of survival after surgery alone. Postoperative RT reduces the risk of local-regional failure and probably improves survival. Patients who are at high risk for recurrence after surgery benefit from more aggressive dose fractionation schedules that may include altered fractionation to decrease the overall time from surgery to the completion of RT. Adjuvant chemotherapy also appears to improve the probability of cure in high risk patients. CONCLUSION: Patients who have a high likelihood of local-regional recurrence after surgery have improved disease control and survival after postoperative RT. PMID- 12579483 TI - Management of labyrinthine fistulae in chronic ear surgery. AB - The appropriate management of labyrinthine fistulae has been debated in the literature for years. After several recent cases of labyrinthine fistulae at our institution, a review of the published data regarding hearing outcome with fistula management was undertaken. Results of this critical review were presented at departmental grand rounds. The grand rounds presentation, data and discussion are presented to better illuminate the topic of labyrinthine fistula management. PMID- 12579484 TI - Marginal mandibulectomy: Oncologic and nononcologic outcome. AB - BACKGROUND AND OBJECTIVES: Marginal mandibulectomy (MM) has been shown to provide an oncologically sound technique with preservation of function and cosmesis. We reviewed our experience with MM and analyzed oncologic and nononcologic complications. PATIENTS AND METHODS: Retrospective review of patients, with clinical and/or radiological evidence of tumor attached to the mandible without cortical erosion, who underwent vertical, horizontal, or oblique marginal mandibulectomy. Data collection included demographics; tumor characteristics; clinical and radiologic relation to the mandible; surgical technique, with attention to neck dissection and facial artery ligation; radiation therapy; osteoradionecrosis (ORN); fractures; and recurrence. RESULTS: Twenty-five patients underwent MM. Primary tumors included oral cavity (22), oropharynx (1). and metastatic neck tumor abutting the inferior/lateral border of the mandible (2). Two patients had local recurrence. Both had close soft tissue margins. Local control rate was 92%. Of 11 patients who had postoperative radiation to the primary site, 4 (36.4%) developed ORN, resulting in fractures in the mandibulectomy site in 2 of them. Two other patients developed early postoperative fractures: 1 patient had had previous radiation, and 1 patient had neck dissections with bilateral facial artery ligation and periosteal stripping. This poor technique led to necrosis of the remaining anterior mandible even before starting radiotherapy. Salvage segmental mandibulectomy was required in both patients. CONCLUSIONS: MM provides an oncologically sound approach to tumors abutting the mandible. Careful attention to preservation of the remaining periosteum and facial arteries will prevent immediate postoperative complications. However, ORN is an important long-term complication that should be taken into account. PMID- 12579485 TI - Ameloblastic carcinoma: report of an aggressive case with multiple bony metastases. AB - Ameloblastic carcinoma is a rare primary tumor of the maxillofacial skeleton with a distinct predilection for the mandible. These lesions may initially show histologic features of ameloblastoma that dedifferentiate over time. Other ameloblastic carcinomas initially present with morphologic features suggestive of ameloblastoma with areas of epithelial dedifferentiation. We herein report a rare case of aggressive ameloblastic carcinoma in a 22-year-old white man who developed widespread bony metastases and expired 4 years after initial diagnosis. PMID- 12579486 TI - Staple-assisted laryngectomy for intractable aspiration. AB - Staple-assisted laryngectomy is a unique method previously reported by Lukyanchenko to prevent wound contamination by using a stapling device for suturing pharyngeal defects in total laryngectomy. We have modified his method and applied it to prevent a postoperative pharyngocutaneous fistula in the treatment of intractable aspiration. In contrast to laryngeal cancer patients, a combined use of an intraluminal light to guide the dissection and laryngofissure to pull the epiglottis can be used to facilitate the use of the stapling device. For most patients with intractable aspiration who have significant malnutrition and drug-resistant bacterial colonization of the pharynx, this method offers certain advantages. This report describes our successful experience with this method in the management of patients with intractable aspiration. PMID- 12579487 TI - Unusual primary and secondary facial blast injuries. AB - PURPOSE: To discuss unusual facial injuries resulting from a bomb blast. MATERIALS AND METHODS: In March 1997, a bomb consisting of a bag of nails was detonated in a coffee shop in Tel Aviv. Two of the wounded were brought to our level 1 Trauma Center with unique facial injuries. Computed tomography (CT) scan and CT angiogram were performed. RESULTS: The blast occurred to the immediate right of the victims who were sitting in an open cafe. Both had tympanic perforation. The first patient showed indirect damage to the facial nerve from a piece of shrapnel located anterior to the carotid artery and medial to the right mandibular angle. The second had a piece of shrapnel lodged in the parapharyngeal space that was initially missed and discovered only on reexamination 3 days later after the patient complained of pain in the temporomandibular joint; there was no facial nerve deficit. The port of entry was probably a small wound in the anterior wall of the external ear canal. CONCLUSIONS: The wounds are probably attributable to the spalling effect of the shrapnel passing through the parotid gland, which has mixed-density tissue. These cases show that nerves are susceptible to damage even in the absence of direct engagement and that the emergency room physician should be alert to even small skin imperfections in blast victims to avoid missing penetrating wounds. PMID- 12579488 TI - [Fibrillin-1 gene mutation in Chinese patients with Marfan syndrome and its gene diagnosis by haplotype analysis]. AB - OBJECTIVE: To analyze fibrillin-1 (FBN(1)) gene mutation in Chinese patients with Marfan syndrome(MFS) and to make a gene diagnosis by haplotype analysis for MFS. METHODS: Nine MFS families were analysed with single strand conformation polymorphism(SSCP) and DNA sequencing. With the use of four primers designed in the flanking sequences of each short-sequence tandem-repeat region in FBN(1) gene, the haplotype-segregation analysis for MFS(B) was performed. RESULTS: In MFS(A)II(1), PCR-SSCP detected SSCP band alterations in exon 25 of FBN(1) gene; direct sequencing showed a small 13bp deletion, the deleted sequence being gcctctgcaccca at base 3243-3456 of cDNA. This mutation caused a frame-shift which was never seen in any unaffected members of the family, and it was a heterozygous mutation; neither of them was identified in 100 chromosomes from 50 normal control individuals. Haplotype-segregation analysis suggested that the disease was passed from Subject I(2) to Subject II(2), Subject II(3), Subject II(5) with the same allele in MFS B family, the proband's daughter also inherited the allele. These data indicated that MFS(B) family was linked to FBN(1) gene, the proband's daughter was an asymptomatic patient. CONCLUSION: The combination of mutation analysis and haplotype analysis can provide more evidence for gene diagnosis. PMID- 12579489 TI - [Association of polymorphisms of vitamin D receptor gene start codon and 3'-end region with bone mineral density in postmenopausal women]. AB - OBJECTIVE: To determine whether vitamin D receptor(VDR) gene start codon polymorphisms and 3'-end region polymorphisms exerted a combined influence on bone mineral density(BMD) in Han postmenopausal women in Beijing area. METHODS: The VDR Fok I and 3'-end region genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism in 110 unrelated postmenopausal women. BMD was measured at the lumbar spine (L(2-4)), femoral neck(Neck), Ward's triangle(Ward's) and trochanter (Troch) using duel-energy X-ray absorptiometry. RESULTS: The frequencies distribution of Fok I, Apa I, Bsm I and Taq I alleles in this cohort all followed the Hardy-Weinberg equilibrium. No significant association of Fok I, Apa I or Taq I genotype with BMD in postmenopausal women was found when these polymorphisms were considered independently, except for Bsm I genotype. When a combined analysis of VDR gene Fok I and 3'-end region polymorphisms was carried out, cross-genotyping Fok I and Apa I polymorphisms was significantly associated with BMD at the L(2-4) (P<0.001), and cross-genotype of Fok I and Taq I was also significantly associated with BMD at the Neck and Troch sites (P<0.05). However, cross-genotyping Fok I and Bsm I polymorphisms was not significantly associated with BMD. Cross-genotyping Apa I and Bsm I or Taq I polymorphisms was not associated with BMD in postmenopausal women, either. CONCLUSION: Although Fok I polymorphisms of VDR gene were not significantly associated with BMD in postmenopausal women, VDR gene Fok I and 3'-region polymorphisms (Apa I and Taq I) had a combined effect on the BMD in postmenopausal women. PMID- 12579490 TI - [Frequencies distribution of GSTT1 and GSTM1 null genotypes in 24 Yunnan populations]. AB - OBJECTIVE: To investigate the frequencies distribution of GSTT1 and GSTM1 null genotypes in 24 Yunnan populations. METHODS: GSTT1 and GSTM1 genotypes were analyzed by PCR procedure. RESULTS: The range of frequencies for GSTT1 and GSTM1 null genotypes in the populations were 0.188-0.633 and 0.400-0.745, respectively, and in the districts were 0.286-0.583, 0.433-0.745 respectively. There was significant relationship between GSTT1 frequencies and populations. CONCLUSION: The frequencies of GSTT1 and GSTM1 null genotypes in 24 Yunnan populations were different, but they were almost the same in different districts of Yunnan. PMID- 12579491 TI - [Cloning and tissue expression analysis of up-regulated cDNA fragment in human gastric cancer]. AB - OBJECTIVE: To identify novel human gastric cancer-associated susceptibility gene for early diagnosis and treatment of gastric cancer. METHODS: A primer was designed for 3'-rapid amplification of cDNA end(RACE) and amplified fragments were cloned, then they were analyzed by sequencing. Compared with ESTs in Genbank, the EST fragment represented a novel gene. Combination of Northern blot and virtual Northern and multiple tissues Northern blot, expression of the cDNA in multiple normal and carcinoma tissues were analyzed. RESULTS: One of the important cDNA bands with poly(A) tail was cloned. This band was named W41. Sequence analysis showed that W41 consists of 533 bp. Basic local alignment search tool analysis revealed that W41 has low identity with any genes from GenBank. This sequence data was submitted to GenBank with accession No. AF 325202. Northern blot revealed that W41 presented higher expression in gastric cancer tissue than in normal tissue. Multiple tissue Northern blot revealed that W41 presented higher expression in multiple cancers than in normal tissues. Virtual Northern revealed that the cDNA presented higher expression in tumor series analysis of gene expression libraries than in normal. CONCLUSION: A novel human gastric cancer-associated cDNA fragment was identified. PMID- 12579492 TI - [Identification of a novel mutation IVS2-2A-->C of SEDL gene in a Chinese family with X-linked spondyloepiphyseal dysplasia tarda]. AB - OBJECTIVE: To identify the mutation of spondyloepiphyseal dysplasia tarda (SEDL) gene in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to make a discussion on the pathogenesis of SEDL at the molecular level. METHODS: In two patients, four exons comprising the SEDL open reading frame as well as their exon/intron boundaries were analyzed by bi-directional direct sequencing of PCR products. The sequencing results were compared against the normal sequences in GenBank to find the mutation. Then the mutation was identified in other members of the family. RESULTS: A nucleotide substitution of the splice acceptor in SEDL intron 2, IVS2 -2A-->C,was detected in two affected individuals (IV(15) V(3)) in the Chinese family with SEDL, but no sequence change occurring on exons 3-6 was detected. The transversion was also identified in four heterozygous carriers. The mutation was not found in two unaffected male individuals and fifteen normal controls. Furthermore, four potential carriers were identified in the family. CONCLUSION: The mutation IVS2 -2A-->C of SEDL gene was firstly determined in the world. The change of the splice acceptor in SEDL intron 2 may cause skipping of exon 3 which is responsible for the disease. Molecular diagnosis can be made by detecting the mutation. PMID- 12579493 TI - [Molecular characterization of a new mutation E122G of human ornithine transcarbamylase gene]. AB - OBJECTIVE: To determine the molecular basis of late onset ornithine transcarbamylase (OTC) deficiency in a Chinese family of Han nationality and the exon sequences of OTC gene of this patient. METHODS: Polymerase chain reaction single strand conformation polymorphism and direct sequencing were used to identify the mutation type. RESULTS: A missense mutation E122G in the conserved residue of exon 4 was identified which is unreported before. CONCLUSION: The E122G mutation in human OTC gene may cause late onset OTC deficiency. PMID- 12579494 TI - [Study on the association of cholesteryl ester transfer protein gene mutations with the susceptibility to coronary atherosclerotic heart disease]. AB - OBJECTIVE: To determine the frequencies of 4 mutations of cholesteryl ester transfer protein (CETP) gene in Chinese population and to investigate the association of the mutations with lipid metabolism and the susceptibility to coronary atherosclerotic heart disease (CHD). METHODS: The target fragments of CETP gene were amplified and analyzed by PCR-restriction fragment length polymorphism technique in 209 unrelated control individuals and 203 CHD patients. The test for Hardy-Weinberg equilibrium was performed using HWE program and statistical analysis was implemented in statistical package SPSS. RESULTS: IVS14A and 451Q mutant genes were not found in either control group or patient group. The frequencies of 405V mutant allele were 0.443 and 0.413 in controls and patients, respectively, while 442G mutant gene frequencies were 0.007 and 0.025, respectively. The observed allele frequencies of I405V and D442G mutation were in accord with Hardy-Weinberg equilibrium. The frequency of 442G mutant gene in patients was significantly higher than that in controls (P=0.043). Compared with the CHD patients without D442G mutation, the 442G heterozygous CHD patients exhibited a significant increase in plasma TC and LDL-C concentration (P=0.017; P=0.041). CONCLUSION: IVS14A and 451Q mutants of CETP gene were rare in Chinese population and 442G mutant gene was possibly one of the susceptibility factors to CHD in Chinese. PMID- 12579495 TI - [Susceptibility gene of systemic lupus erythematosus in 16q12 in Chinese cohort]. AB - OBJECTIVE: To localize susceptibility loci in Chinese systemic lupus erythematosus (SLE) cohort by linkage disequilibrium mapping the genomic interval in human chromosome 16 so as to understand whether the pathogenesis of human SLE is related to chromosome 16. METHODS: Five microsatellite markers in chromosome 16 spanning from 57.79 cM to 65.1 cM were used for fluorescence based genotyping in 157 SLE families. Evidence for linkage disequilibrium was assessed using the extended transmission disequilibrium test (ETDT) program and Genehunter software. Susceptibility gene was searched in the positive locus, and real-time PCR method was used to detect gene expression. RESULTS: With the ETDT, evidence for linkage disequilibrium of the marker D16S409 (P=0.0278) and D16S517 (P<0.0001) with SLE were found. It was observed that 271 bp allele of D16S517 was much in evidence for preferential transmission, while 277 bp allele was found to be transmitted less often than expected. Genehunter analysis is concordant with ETDT. By real time PCR, OAZ(OLF1/ EBF-associated zinc finger protein) gene which is located in the positive locus showed higher expression in SLE patients than in normal controls. CONCLUSION: Results reveal that 16q12 (58.46 cM) is associated with Chinese SLE. OAZ is a likely susceptibility gene in this locus for SLE. PMID- 12579496 TI - [The expression and imprinting status of insulin-like growth factor 2 gene in colorectal cancer]. AB - OBJECTIVE: To study the imprinting status and expression level of insulin-like growth factor 2 (IGF2) gene in colorectal cancer and to provide a clue for the mechanism of carcinogenesis of colorectal cancer. METHODS: The expression levels of IGF2 in the paired colorectal cancer and adjacent normal tissue were examined and compared by use of semi-quantitative reverse transcription-polymerase chain reaction. The imprinting status of IGF2 was detected by restriction fragment length polymorphism. The relationships between the expression level of IGF2, its imprinting status, and the carcinogenesis of colorectal cancer were analyzed. RESULTS: IGF2 was overexpressed in 82.4% (28/34) of colorectal cancer tissues which was significantly higher than those of the matched normal tissues (P<0.01, t=3.01). 87.5% (14/16) of colorectal cancer showed loss of imprinting(LOI), while 71.4%(10/14) of normal tissues also displayed LOI of IGF2. CONCLUSION: Overexpression of IGF2 was found to play an important role in carcinogenesis of colorectal cancer. LOI of IGF2 may be a prophase manifestation of colorectal cancer. PMID- 12579498 TI - [Association of coagulation factor V, VII gene polymorphisms with coronary heart disease]. AB - OBJECTIVE: To study the distribution of coagulation factor V(FV), VII(FVII) gene polymorphisms in Chinese Han population and the association of the polymorphisms with coronary heart disease(CHD). METHODS: Genotypes of FV FVII were typed in 234 CHD patients and 210 controls by polymerase chain reaction-restriction fragment length polymorphism. Selected coronary angiography was performed in 234 CHD patients. RESULTS: FVII allelic frequencies of R,Q and H7,H6 were 94.6%, 5.6%, 70.3%, 29.7% and 91.9%, 8.1%, 60.9%, 39.1% in CHD group and control group respectively. Genotype distribution was in accordance with Hardy-Weinberg equilibrium. There was no significant difference in frequencies of allele and genotype in R353Q or HVR4 polymorphisms between CHD group and control group. The distribution of allele and genotype in R353Q was of significant difference between non-myocardial infarction subgroup and myocardial infarction subgroup (chi2 = 4.711, P<0.05, OR=0.37,95% CI: 0.15-0.94). However, HVR4 polymorphism was not found to be of significant difference within two group (chi2 = 0.142, P>0.05). There was no FV Leiden mutation in all the CHD patients and normal controls. CONCLUSION: The Q allele of the R353Q polymorphism of the FVII gene may be a protective factor against myocardial infarction. PMID- 12579497 TI - [Association of genetic polymorphisms in the DNA repair gene XPD with risk of lung and esophageal cancer in a Chinese population in Beijing]. AB - OBJECTIVE: XPD polymorphisms at Asp312Asn and Lys751Gln sites have been shown to modulate DNA repair capacity. The authors therefore assessed the relationship between these XPD polymorphisms and susceptibility to lung and esophageal cancer in a Chinese population via a hospital-based, case-control study. METHODS: Genotypes were determined by PCR-restriction fragment length polymorphism approaches in 383 healthy controls, 351 patients with lung cancer, and 325 patients with esophageal squamous cell carcinoma (SCC). The adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated using multivariate logistic regression. RESULTS: Individuals carrying at least one 312Asn variant allele (Asp/Asn and Asn/Asn genotypes) were at an increased risk for lung SCC as compared with those with the Asp/Asp genotype (OR 1.80; 95% CI: 1.10-2.93; adjusted for age, sex and smoking), but this increased risk was not observed among patients with adenocarcinoma of the lung (adjusted OR: 1.07; 95% CI: 0.55 2.08). Furthermore, stratified analysis indicated a multiplicative interaction between tobacco smoking and the variant XPD 312Asn and 751Gln alleles on risk of lung SCC. The ORs of lung SCC for the variant XPD 312Asn and 751Gln alleles with smoking>or=29 pack/year were 12.44 (95% CI: 4.97-31.17) and 10.74 (95% CI: 4.51 25.57), respectively. No significant association between the Asp312Asn or Lys751Gln polymorphism and the risk of esophageal cancer was found. CONCLUSION: The above findings indicate that the Asp312Asn and Lys751Gln polymorphisms in the XPD locus are associated with the risk of lung SCC but not lung adenocarcinoma or esophageal SCC in this Chinese population. PMID- 12579499 TI - [Analysis on mutation of adrenoleukodystrophy gene in exon 1 and exon 5]. AB - OBJECTIVE: To elucidate the molecular mechanism of X-linked adrenoleukodystrophy(ALD) in Chinese. METHODS: Polymerase chain reaction in exon 1, exon 5 and their flanking sequences and direct DNA sequencing of ALD gene were performed in four patients, their mothers and twenty normal individuals as controls. RESULTS: A splice mutation was identified in the interface of exon 5 and intron 5 (1875 G-->A). This splice mutation in 5' end of intron 5 might lead to abnormal splice in exon 5 and exon 6 and bring about unstable and abnormal ALD protein; the lignoceryl CoA ligase could not transport very long chain fatty acids (VLCFA) into peroxisome and could not function normally; consequently, defective beta-oxidation of VLCFA in peroxisome could result in an accumulation of VLCFAS in the central nervous system, adrenal gland and blood. CONCLUSION: The splice mutation in 5' end of intron 5 leading to abnormal splice in exon 5 and exon 6 appears to be one of the causes of X-linked recessive adrenoleukodystrophy. PMID- 12579500 TI - [Y-chromosome-specific microsatellite variation in Li ethnic groups of Hainan Island, China]. AB - OBJECTIVE: The study was conducted to reveal the distribution of genetic polymorphism of four Y chromosome specific short tandem repeat (Y-specific STR) loci in Li ethnic groups in Hainan Island, China. METHODS: Four tetranucleotide STR loci were simultaneously amplified with fluorescently labeled primers, and genotypes were determined with an automated DNA sequencer. RESULTS: Among 230 unrelated males, the alleles at the four Y-specific STR loci were composed of some complex repeat structure. 4,5,4,5 alleles were observed in loci DYS3891, DYS390, DYS391, DYS393 respectively. A set of human allele ladders for the typing of the four Y-specific STRs was obtained in Li ethnic population. Gene diversity index (D) and haplotype diversity data were estimated for the four Y-STRs. CONCLUSION: The preliminary study indicates a reference population for detecting male migration events and should be useful in population genetics and forensic applications. PMID- 12579501 TI - [Screening and identification for cDNA of differentially expressed genes in human primary hepatocellular carcinoma]. AB - OBJECTIVE: Screening and identification of differentially expressed genes in human primary hepatocellular carcinoma(HCC). METHODS: The differentially expressed genes subtracted cDNA library of HCC constructed by suppression subtractive hybridization(SSH) technique was screened by colony in situ hybridization, then the positive clones were further screened with PCR amplification. The positive clones were sequenced and analyzed for homology in the Genbank databases with Basic Local Alignment Search Tool BLAST . The novel cDNA sequences were analyzed by Northern blot analysis. RESULTS: Thirteen positive clones were obtained, and 11 cDNA sequences were identified. Sequences of 11 cDNA showed that 6 cDNA were homologous with the genes published in Genbank and 5 cDNA were unknown genes. Northern blot indicated that 3 novel cDNA(>300 bp) were only expressed in HCC. CONCLUSION: The subtracted cDNA library constructed by SSH technique contains differentially expressed genes of HCC. Three novel cDNA sequences might be differentially expressed genes of HCC. Further screening the library and gaining the whole gene sequence may lay a foundation for identifying differentially expressed genes in HCC. PMID- 12579502 TI - [Application of homozygosity mapping to the fine mapping of the osteoporosis pseudoglioma syndrome locus]. AB - OBJECTIVE: To evaluate the role of homozygosity mapping in the fine mapping of the genes responsible for the rare autosomal recessive diseases. METHODS: Polymerase chain reaction-single sequence length polymorphism was used to genotype the family members from 8 families with osteoporosis-pseudoglioma syndrome(OPS) for 14 polymorphic loci within candidate region. The OPS candidate region was narrowed by searching for homozygous region in affected. RESULTS: The OPS candidate region was narrowed to a 1 cM interval between D11S1296 and D11S4136. CONCLUSION: Homozygosity mapping is a powerful method for mapping and narrowing the candidate region of the genes responsible for the rare autosomal recessive diseases. PMID- 12579503 TI - [An allelotype study of primary and corresponding recurrent glioblastoma multiforme]. AB - OBJECTIVE: To investigate molecular genetic alterations associated with primary and corresponding recurrent glioblastoma multiforme(GBM) and to identify which chromosomal regions of the whole genome may be involved in the recurrence of primary GBM. METHODS: A high-resolution allelotyping study of one patient's primary GBM and corresponding recurrent GBM was performed by PCR-based loss of heterozygosity(LOH) analysis with the use of 382 fluorescent dye-labeled polymorphic microsatellite markers covering all 22 autosomes. The mean genetic distance between two flanking markers is 10 cM. RESULTS: LOH at locus D9S157 on 9p21 and at loci D10S537, D10S185, D10S192, D10S597, D10S587, D10S217 on 10q21.3 26.3 was observed in the primary GBM. As for corresponding recurrent tumor, LOH was observed not only in expanded regions on 9p21 and 10q21.3-26.3 but also on multiple other chromosomal arms, including 1q, 7p,7q, 21q, 20p, 20q, 10p, 19p, 19q. CONCLUSION: Chromosome 9p and 10q may be involved in the development of this GBM. Although histopathological diagnoses of the primary and corresponding recurrent tumor are identical, the recurrence of GBM is characterized by an increased involvement of molecular genetic abnormalities and may be accompanied by inactivation of more tumor suppressor genes. PMID- 12579504 TI - [Detection of chromosome 8 anomalies in ovarian carcinoma by fluorescence in situ hybridization]. AB - OBJECTIVE: To detect the relationship between chromosomal anomalies and the pathogenesis, development and prognosis of ovarian carcinoma. METHODS: Thirty-six specimens of ovarian carcinoma (n=12), ovarian benign tumor (n=12), and normal ovary (n=12) were examined by fluorescence in situ hybridization (FISH). RESULTS: Twelve cases of mutations, including trisomy 8, monosomy 8 or tetraploid 8 chromosomal anomalies, were found in the group of ovarian carcinoma, making up 100% (12/12). Three cases of trisomy 8 chromosomal anomalies were found in the group of ovarian benign tumor, accounting for 25% (3/12). No anomaly was found in the normal group. There were significant differences between the three groups, P<0.001. CONCLUSION: The above anomalies of chromosome 8 are significantly associated with the pathogenesis and development of ovarian carcinoma. The anomalies may occur in the early stage of the carcinoma, and may be significantly associated with the pathological differentiation and clinical stage of the case. PMID- 12579505 TI - [Preliminary analysis on human telomerase reverse transcriptase gene expression in acute leukemia]. AB - OBJECTIVE: To explore the relationship between gene expression of human telomerase reverse transcriptase (hTERT) and its clinical characteristics in leukemia. METHODS: The protocol of RT-PCR was used to detect the hTERTmRNA expressing levels in peripheral blood samples from leukemic patients under primary treatment(n=42), in complete remission(n=21), with recurrent leukemia (n=4); and from normal subjects (n=5), respectively. RESULTS: The positive percentage of hTERTmRNA expression was 73.81% for the primary treatment cases, and 19.05% for the complete remission cases. All of the recurrent cases gave positive results. One of the normal controls presented low level of hTERTmRNA expression. The expressing level of hTERTmRNA in primary treatment cases was 0.64+/-0.21, in complete remission leukemia 0.31+/-0.16, in recurrent cases 0.84+/-0.09, and in normal controls 0.10. CONCLUSION: The activation of telomerase may be an essential factor in the development of leukemia and usually be the late event in its progression. As an indicator of leukemia cell, the detection of hTERT mRNA may be used in clinical analysis, disease monitoring and prognosis judgement. PMID- 12579506 TI - [Sex determination of human preimplantation embryo using nested polymerase chain reaction]. AB - OBJECTIVE: Using nested polymerase chain reaction (PCR) to perform preimplantation gender diagnosis. METHODS: One (or two) lymphocyte and blastomere (n=50/group) were collected and prepared under the following conditions: (1) water only (H(2)O); (2) freeze-thaw liquid nitrogen, then boiling; (3) potassium hydroxide/dithiotheriol, heated to 65 degree centigrade, followed by acid neutralization (KOH). Cells were analyzed by PCR using nested primers amplification with amelogenin gene. RESULTS: The amplification rate and allele dropout (ADO) rate for male lymphocytes by the three methods were 83%, 94%, 95% and 24%, 12%, 4%, respectively. Using two cells per reaction did not increase the amplification rate for the KOH method. CONCLUSION: The KOH method for DNA preparation is superior to the other methods evaluated. Dual blastomere biopsy and independent blastomere analysis may improve preimplantation diagnostic reliability. PMID- 12579507 TI - [Apolipoprotein E genotypes and carotid artery atherosclerosis in type 2 diabetes mellitus]. AB - OBJECTIVE: To investigate the relationship between carotid artery intima media thickness (IMT) and apolipoprotein (Apo) E gene polymorphisms in type 2 diabetes mellitus (DM2). METHODS: Two hundred and fifty-five DM2 patients without angiopathy and 107 healthy individuals were selected. PCR/allele-specific oligonucleotide probe was used to determine their apoE genotypes. RESULTS: The prevalence distribution of apoE genotypes and alleles in DM2 patients and that in controls were similar. The TC, LDL-C and Lp(a) concentrations in e4/4, e4/3 subgroups were significantly higher than those in e3/2, e2/2 subgroups (P<0.05). The average value of IMT in e4/4 e4/3 carriers (0.89 mm) was significantly greater than that in e3/2 e2/2 carriers (0.62 mm) (P<0.05). After adjustment for TC, LDL-C, TG, Lp(a), FBG, HbA1c, age, BMI, and smoking, ANCOVA showed that the average value of carotid IMT was significantly greater in subjects with e4/4 e4/3, compared with that in subjects with e3/2 e2/2(P=0.033). CONCLUSION: Apo e4 allele increases the risk for carotid artery atherosclerosis in the early stage of diabetic population. PMID- 12579508 TI - [Associations between six functional genes and schizophrenia]. AB - OBJECTIVE: To assess the associations between schizophrenia and six functional genes: dopamine D2 receptor gene (DRD2), dopamine D4 receptor gene (DRD4), 5 hydroxytryptamine 2A receptor gene (5-HT2A), 5-HT6 receptor gene (5-HT6), catechol-O-methyltransferase gene (COMT) and dopamine transporter gene (DAT1). METHODS: With the techniques of Amp-RFLP and Amp-FLP, association analysis was made between schizophrenia and the six genes in 67 schizophrenic patients from Chinese Han population. RESULTS: (1) Neither genotypes nor alleles of DRD2, 5 HT2A, 5-HT6 and COMT gene showed significant differences between patients and controls (P>0.05). (2) Six repeats (6R) in DRD4 gene, the allele of 480 bp and the genotype of 480/520 in DAT1 gene were found to be of significant differences between the two groups (P<0.05). (3) Only one negative association was observed between the 480 bp allele of DAT1 gene and schizophrenia (OR=0.441, 95% CI:0.202 0.963, Z=2.05, P<0.05). CONCLUSION: The 480 bp allele of DAT1 gene is negatively associated with schizophrenia in Chinese Han population, which stands for the dopamine hypothesis of schizophrenia. PMID- 12579509 TI - [Single nucleotide polymorphism loci of HIV-1 coreceptor CCR5 gene in Chinese Han people]. AB - OBJECTIVE: To investigate the single nucleotide polymorphism(SNP) loci of HIV-1 coreceptor CCR5 gene in Chinese Han people. METHODS: The coding region of CCR5 was amplified using 2 pairs of primers and the PCR products of all 42 healthy subjects were sequenced by 4 different primers. The results of sequencing were analyzed by DNAstar in search of SNP loci. RESULTS: Six SNP loci were discovered in the coding region of CCR5, among them four SNPs, i.e. 184A-->G, 503G-->T, 688G ->A and 999G-->T, cause amino acids changes and two SNPs are nonsense mutations. One cytosine deletion at the 894nt results in frame shift mutation and prematured termination. 184A-->G, 503G-->T and 999G-->T were found in Chinese Han people for the first time. The allelic frequencies of mutant 184G, 503T and 999T alleles were 1.1%, 21.1% and 10.0% in healthy Hans, respectively. The population distribution of G503T markedly deviated from Hardy-Weinberg equilibrium. CONCLUSION: The SNP loci in the coding region of CCR5 in Chinese Han people has its own characteristics, which is not consistent with those of Japanese and obviously different from those of Caucasian and African. PMID- 12579510 TI - [Expression of human anti-apoptotic gene survivin and its splice in normal human gastric tissue and gastric cancer]. AB - OBJECTIVE: To analyze the expression of human anti-apoptotic gene survivin (SVV) in normal human gastric tissues and gastric cancer. METHODS: SVV cDNA clone was obtained from human gastric cancer tissues by virtue of RT-PCR, using Dig-marked cRNA probe in situ hybridization to analyze its expression in normal human gastric tissues and gastric cancer. RESULTS: Two SVV cDNA clones, SVV-S4A and SVV S1B were obtained. The sequence of the former is identical to that of the well known SVV cDNA; however, in the sequence of the latter, the third exon was missed, i.e., there are only two exons in SVV-S1B. In situ hybridization showed that SVV-S4A is mainly expressed in gastric cancer tissues whereas SVV-S1B is mostly expressed in normal gastric tissues. CONCLUSION: There is difference between SVV-S4A and SVV-S1B in respect to their characteristics of expression in gastric cancer and normal gastric tissues. PMID- 12579511 TI - [Gene polymorphisms in Uighur patients with Abnormal Savda]. AB - OBJECTIVE: To investigate gene polymorphisms in Uighur patients with Abnormal Savda. METHODS: Polymorphisms of genomic fingerprints generated by arbitrarily primed PCR(AP-PCR) can be used for distinguishing between strains of almost any organism. The authors of this article applied the technique to the polymorphism analysis on the DNA of Uighur patients suffering from four kinds of Abnormal Savda (n=116), and on the DNA of a control group(n=50). RESULTS: From the patients, 42 fragments were obtained, among them 27(64.2%) fragments did not show gene polymorphisms. On the 220-344 bp regions, the authors detected 6 AP-PCR products showing individual specificity. CONCLUSION: The polymorphisms may serve as a molecular marker for gene diagnosis in the patients with Abnormal Savda. PMID- 12579512 TI - [Association between HLA-DRB1, DQB1 genes and pemphigus vulgaris in Chinese Hans]. AB - OBJECTIVE: To investigate the predisposing role of HLA-DRB1, DQB1 genes in pemphigus vulgaris (PV). METHODS: Polymerase chain reaction-specific sequence primers method was used to type HLA-DRB1, DQB1 subregion in the patients with PV of Han nationality from Jiangsu and Anhui provinces and matched control subjects. RESULTS: DR4, DRB1*14(*1401,*1404,*1405) gene frequencies in PV patients were significantly higher than those in controls (Pc<0.05 and Pc<0.01 respectively). DQB1*0302, DQB1*0503 gene frequencies were significantly higher in PV patients (Pc<0.05). Further typing of DR4 positive subjects revealed that the gene frequencies of DRB1*0406, *0403 were significantly increased in PV patients as compared with controls (Pc<0.05). The haplotype frequencies of HLA-DRB1*04, DQB1*0302 and HLA-DRB1*14, DQB1*0503 in PV patients were significantly higher than those in controls (P<0.05). CONCLUSION: The results suggest that the combination of HLA-DRB1*4, DQB1*0302 and HLA-DRB1*14, DQB1*0503 forms putative susceptible haplotypes for PV patients in Chinese Hans. PMID- 12579513 TI - [Genetic polymorphism of 3 STR loci of CSF1PO, TPOX and TH01 in Chinese Sibo population]. AB - OBJECTIVE: To obtain allele and genotype frequencies and related forensic data of CF1PO, TPOX and TH01 loci in Chinese Xinjiang Sibo population. METHODS: Genomic DNA from peripheral blood mononuclear cells of normal Chinese Xinjing Sibo population was used as template, and CSF1PO, TPOX and TH01 fragments were amplified by PCR. The PCR products were analyzed by 4% denaturing PAGE and detected using silver stain detection. RESULTS: Nine alleles were found at CSF1PO locus, eight alleles at TPOX locus and eight alleles at TH01 locus in Chinese Sibo population. All the 3 loci complied with Hardy-Weinberg equilibrium. The heterozygosities were 0.9426, 0.8361 and 0.8853, and the polymorphism information contents were 0.8298, 0.7213 and 0.7626 for CSF1PO, TPOX and TH01, respectively. CONCLUSION: The data on the alleles frequency of these 3 STR loci might be used for individual identification and paternity identification and for genetic researches in Chinese Sibo population. PMID- 12579514 TI - [Gene frequencies of 4 STR loci in Tibetan population of Yunnan province]. AB - OBJECTIVE: To investigate the gene frequencies of 4 STR loci in Tibetan population of Yunnan. METHODS: Multiple polymerase chain reaction (PCR), denaturing polyacrylamide gel electrophoresis and silver staining were used to detect D21S11, D8S1179, D16S539 and LPL loci. DNA samples collected from 105 unrelated Tibetan individuals in Yunnan province were analyzed. RESULTS: At D21S11, D8S1179, D16S539 and LPL loci, 13, 8, 7, 6 alleles and 33, 21, 16 and 9 genotypes were observed, respectively. The genotype distribution of the 4 STR was in accordance with the Hardy-Weinberg equilibrium. CONCLUSION: The high combined discrimination power and exclusion power of the four loci in Tibetan population make multi-PCR detection a valuable tool for forensic identity, genetics and anthropology. PMID- 12579515 TI - Glutamine and glutamate--their central role in cell metabolism and function. AB - Glucose is widely accepted as the primary nutrient for maintenance and promotion of cell function. However, we propose that the 5-carbon amino acids, glutamine and glutamate, should be considered to be equally important for maintenance and promotion of cell function. The functions of glutamine are many and include: substrate for protein synthesis, anabolic precursor for muscle growth, acid-base balance in the kidney, substrate for ureogenesis in the liver, substrate for hepatic and renal gluconeogenesis, an oxidative fuel for intestine and cells of the immune system, inter-organ nitrogen transport, precursor for neurotransmitter synthesis, precursor for nucleotide and nucleic acid synthesis and precursor for glutathione production. Many of these functions are connected to the formation of glutamate from glutamine. We propose that the unique properties regarding concentration and routes of metabolism of these amino acids allow them to be used for a diverse array of processes related to the specialized function of each of the glutamine utilizing cells. In this review we highlight the specialized aspects of glutamine/glutamate metabolism of different glutamine-utilizing cells and in each case relate key aspects of metabolism to cell function. PMID- 12579516 TI - Evidence that direct DNA uptake through cut shoots leads to genetic transformation of Solanum aviculare Forst. AB - The reporter genes GUS, NPTII and BAR, either separately or in combination, have been exploited to determine if DNA which can directly enter plants, circulate within the plant and enter nuclei, can also integrate into the genome in a manner which will permit gene expression. Feeding of either seed-derived or adventitious cut shoots of Solanum aviculare with the GUS gene followed by rooting of the shoots and growing on, resulted in all tissues of the plant showing GUS activity as detected cytochemically. Southern blot analysis of plants derived from the adventitious shoots confirmed the presence of the reporter gene in roots. Reporter gene expression was observed also in the F1 generation. If GUS and NPTII or GUS, NPTII and BAR were fed together, then in each case it was possible to have both expression and Southern blot confirmation of each of the genes. There was a relatively high rate of transformation of approximately 5% of the fed stems across all experiments conducted during the present study. PMID- 12579517 TI - Effects of steroid hormones on five functional parameters of Tetrahymena: evolutionary conclusions. AB - The unicellular Tetrahymena pyriformis was studied for chemotaxis, chemotactic selection, phagocytosis, growth and body shape changes in the presence of water soluble (beta-cyclodextrin-coupled) steroid hormones (testosterone, estradiol, progesterone, hydrocortisone and dexamethasone). Testosterone was chemoattractant over a wide range of concentrations, while progesterone and dexamethasone were active only at one concentration (10(-5) and 10(-6) mg ml(-1) respectively) and were either neutral or repellent at other concentrations. Hydrocortisone and estradiol were unambiguously chemorepellent. Chemotactic selection enhanced the effect of testosterone and estradiol, while in the case of hydrocortisone the action was reversed. The other parameters were mildly influenced by the steroid hormones. The results call attention to the fine molecular recognition capacity of Tetrahymena and to the possible rapid effects of steroid hormones at membrane receptors at a very low evolutionary eukaryotic level. PMID- 12579518 TI - Effect of cobalt on the oxidative status in heart and aorta of streptozotocin induced diabetic rats. AB - Effects of cobalt on the antioxidant status of control and streptozotocin diabetic rat heart and aorta were examined at the second, fourth and sixth week of treatment. Rats were divided into four groups: control, diabetic, control treated with cobalt chloride and diabetic treated with cobalt chloride. Diabetes was induced by tail vein injection of streptozotocin (STZ). Cobalt treatment groups were given 0.5 mM of CoCl(2) in drinking water. The rats in both groups were further subdivided into three groups of six rats each. Rats in these subgroups were studied at 2-week intervals up to 6 weeks. At the end of the experiment, all animals were sacrificed by decapitation, heart and aorta samples were removed for determination of thiobarbituric acid reactive substance (TBARS) level and superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities. It was found that lipid peroxidation levels and antioxidant enzyme activities were increased in the streptozotocin-induced diabetic rats at all times studied. Cobalt treatment of diabetic rats (0.5 mM in drinking water) resulted in attenuation of the increased levels of TBARS and antioxidant enzyme activities in heart and aorta. Thus, the effect of oral administration of cobalt at this dose during the early stage of experimental diabetes can be considered as a consequence of altered endogenous defence mechanisms in heart and aorta. PMID- 12579519 TI - Melatonin effect on renal oxidative stress under constant light exposure. AB - Recently, numerous studies have shown antioxidant actions of melatonin. Melatonin at both physiological and pharmacological levels stimulates glutathione peroxidase, glutathione reductase and superoxide dismutase activities in the brains of rats and chickens. This study was designed to evaluate the effect of melatonin on nephropathy and oxidative stress under constant light exposure. Nephropathy was induced by adriamycin administered in a single dose (25 mg kg(-1) b.w., i.p.). Melatonin was injected i.p. (1,000 microg kg(-1) b.w./day). Malondialdehyde, reduced glutathione, glutathione peroxidase, glutathione reductase, glutathione transferase, catalase and superoxide dismutase were determined in kidney. Urea, creatinine and total proteins in plasma and proteinuria were evaluated and melatonin was determined. Results show a decrease in melatonin levels. Similar effects occurred with the antioxidant enzyme activities and reduced glutathione. Likewise, adriamycin and constant light induced significant enhancement of malondialdehyde. All changes induced both by adriamycin and constant light were reverted to normal by melatonin administration. Constant light exposure was associated with an increase in oxidative stress and nephropathy induced by adriamycin. Treatment with melatonin decreased lipid peroxides, and permitted a recovery of reduced glutathione, scavenger enzyme activity and parameters of renal function. PMID- 12579520 TI - Cellular localization of integrin isoforms in phenylephrine-induced hypertrophic cardiac myocytes. AB - Cardiac hypertrophy is characterized by remodeling of the extracellular matrix (ECM). Integrins are cell-surface molecules that link the ECM to the cellular cytoskeleton where they play roles as signaling molecules and transducers of mechanical force. To clarify the possible roles of integrins in cardiac myocyte hypertrophy, we investigated the cellular localization and expression of ECM proteins and integrins in both normal cardiac myocytes and phenylephrine-induced hypertrophic myocytes. Addition of phenylephrine (PE) to cultured neonatal cardiac myocytes induced sarcomeric organization, increase in cell size, and synthesis of the hypertrophic marker, atrial natriuretic factor (ANF). In particular, fibronectin and collagen underwent dramatic localization changes during PE-induced cardiac hypertrophy. Significant changes were noted in the cellular localization of the respective collagen and fibronectin receptors, integrin alpha1 and alpha5, from diffuse to a sarcomeric banding pattern. Expression levels of integrins were also increased during hypertrophy. Treatment with okadaic acid (OA), an inhibitor of protein phosphatase 2A (PP2A), resulted in inhibition of hypertrophic response. These results suggest that dephosphorylation of integrin beta1 may be important in the induction of cardiac hypertrophy. PMID- 12579521 TI - Effects of erythropoietin and pentoxifylline on the oxidant and antioxidant systems in the experimental short bowel syndrome. AB - In this study, we investigated the effects of erythropoietin (Epo), and pentoxifylline (Ptx) on the oxidant and antioxidant systems in the experimental short bowel syndrome. Spraque-Dawley rats were divided into four groups and all animals underwent 75% small bowel resection. Group E was treated with 500 IU kg(- 1) Epo subcutaneously (s.c.), group P with 50 mg kg(- 1) day(- 1) s.c. Ptx and group E+P with 500 IU kg(- 1) s.c. Epo plus 50 mg kg(- 1) day(- 1) s.c. Ptx for a period of 28 days. In group C, which is the control group, no drug treatment was given. At the end of 28 days the experimented rats were killed and ileum samples excised for biochemical and histopathological testing. Malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels were determined in ileum homogenates. When compared to group C, the MDA and GSH-Px levels were significantly decreased (p < 0.05), but SOD activity was not changed (p > 0.05) in groups P and E+P, whereas both MDA and SOD and also GSH-Px activities were not changed significantly in group E (p > 0.05). The average villous length, crypt depth, muscular thickness and mucosal length were measured in all groups. The average crypt depth and mucosal length were statistically higher in the group P than group C (p < 0.001, p < 0.01, respectively). In addition, the crypt depth was statistically higher in both E and E+P groups as compared to group C (p < 0.001, p < 0.01, respectively). Therefore, our study indicates that Ptx may be more effective than Epo in reducing lipid peroxidation. Moreover, we considered that Ptx may give this protective effect by inhibiting the free oxygen radicals to a greater extent than developing the antioxidant capacity. PMID- 12579522 TI - Pathophysiological role of nitric oxide and adrenomedullin in autism. AB - Several studies indicate that nitric oxide (NO) is involved in the aetiopathogenesis of many neuropsychiatric disorders such as schizophrenia, bipolar disorder, depression, Alzheimer's disease, Hungtington disease and stroke. Although it has not been investigated yet, several recent studies proposed that NO may have a pathophysiological role in autism. Adrenomedullin (AM), a recently discovered 52-amino acid peptide hormone, induces vasorelaxation by activating adenylate cyclase and also by stimulating NO release. AM immune reactivity is present in the brain consistent with a role as a neurotransmitter. It has been stated that NO and AM do function in the regulation of many neurodevelopmental processes. We hypothesized that NO and AM activities have been affected in autistic patients and aimed to examine these molecules. Twenty-six autistic patients and 22 healthy control subjects were included in this study. AM and total nitrite (a metabolite of NO) levels have been measured in plasma. The mean values of plasma total nitrite and AM levels in the autistic group were significantly higher than control values, respectively (p < 0.001, p = 0.028). There is no correlation between total nitrite and AM levels (r = 0.11, p = 0.31). Certainly, this subject needs much further research investigating autistic patients in earlier periods of life and with subtypes of the disorder. PMID- 12579523 TI - ATP-induced calcium oscillations and change of P2Y subtypes with culture conditions in HeLa cells. AB - ATP, UTP, ADP and UDP induced intracellular Ca(2+) responses and oscillations in HeLa cells that sometimes lasted over 1 h. The response is due to the activation of P2Ys, G-protein coupled ATP receptors, because the oscillations persisted for several minutes even in Ca(2+)-free solution, and suramin and PPADS, antagonists of ATP receptors, partially inhibited the response. The potency of these nucleotides varied with the culture or cell conditions, i.e. UTP was generally most potent but in some cases UDP was more potent; responses to UDP were variable while those to ATP were constant. In addition, Ca(2+) responses to ATP and UDP were additive. These findings suggested the existence of two or more subtypes of P2Ys in HeLa cells. RT-PCR experiments revealed the existence of P2Y(2), P2Y(4) and P2Y(6). Recovery from starvation (culture in FBS-free medium overnight and re addition of FBS) increased the responses to UTP and UDP but not to ATP, suggesting that the number or activity of P2Y(6) and/or P2Y(4) receptors may increase with cell proliferation in HeLa cells. PMID- 12579524 TI - Effect of cigarette smoke on lipid peroxidation, antioxidant enzymes and NMDA receptor subunits 2A and 2B concentration in rat hippocampus. AB - The effect of cigarette smoke on lipid peroxidation and antioxidant enzymes such as catalase, superoxide dismutase, glutathione peroxidase and on the concentration of N-methyl-d-aspartate receptor (NMDAR) subunits 2A and 2B in the hippocampus of Sprague-Dawley rats exposed to cigarette smoke for 2h/day for a period of 4 weeks was determined. It was observed that NMDAR 2A and 2B concentrations in the hippocampus were enhanced in the case of animals exposed to cigarette smoke, whereas lipid peroxidation and antioxidant enzyme activities did not show any change as compared to control animals. The results of our study suggest that cigarette smoke induces NMDAR 2A and 2B expression in the hippocampus, and that this is not due to an increased lipid peroxidation, because cigarette smoke has no effect on lipid peroxidation and antioxidant enzyme activities in the hippocampus. PMID- 12579525 TI - Glutathione redox system, GSH-Px activity and lipid peroxidation (LPO) levels in tadpoles of R.r.ridibunda and B.viridis. AB - Total glutathione (t-GSH), reduced glutathione (GSH), glutathione disulphide (GSSG) levels, t-GSH/GSSG ratio, glutathione peroxidase (GSH-Px) activity and lipid peroxidation (LPO) levels were investigated during the development period of a predominantly aquatic amphibian R.r.ridibunda and a predominantly terrestrial amphibian B. viridis. While t-GSH and GSH showed a similar trend, GSSG concentration increased significantly (p<0.05) during the larval stages in R.r.ridibunda larvae. In contrast to R.r.ridibunda larvae, there was no significant (p>0.05) change between 1 and 5 weeks in the t-GSH and GSH concentrations of B. viridis. t-GSH and GSH concentrations of B. viridis larvae became sharply elevated after the fifth week, GSSG levels increased 3.25-fold during the metamorphosis. The t-GSH/GSSG ratio fluctuated and the lowest t GSH/GSSG ratios were observed at the third week for both species. GSH-Px activities for both species increased significantly (p<0.05) during the growing period. The highest GSH-Px activities in R.r.ridibunda and B.viridis were observed at the eighth week and they were 3.45 +/- 0.17 and 4.1 +/- 0.21 IU mg( 1), respectively. The membrane LPO levels in the R.r.ridibunda and B. viridis tadpoles significantly (p<0.001) decreased from 206 +/- 10.3 to 146 +/- 7.3 and from 198 +/- 9.9 to 23 +/- 1.15 nmol MDA g(-1) w.w., respectively. PMID- 12579526 TI - Human cutaneous melanoma expresses a significant phosphate-dependent glutaminase activity: a comparison with the surrounding skin of the same patient. AB - The protein content and the activity and type of phosphate-dependent glutaminase were determined in freshly pigmented lesions obtained from human melanoma and adjacent skin. Significant phosphate-dependent glutaminase activity was found in both the melanoma and non-pigmented adjacent skin areas. A comparison between the pigmented and adjacent skin areas suggests the occurrence of gradual metabolic changes that result in an increased protein content in the centre of the neoplasia. The presence of a kidney-type glutaminase (K(m) of 2-5 mm) indicates a high sensitivity of the melanoma to variations in glutamine plasma levels (0.6 to 1 mm). These data lead us to postulate that glutamine supply is an important factor for melanoma cell proliferation, being a source of nitrogen for DNA and RNA synthesis. The intense neovascularization observed in melanoma ensures the oxygen supply that is required for glutamine oxidation. These findings support the proposition that glutamine is an important fuel for melanoma. PMID- 12579527 TI - Effect of arginine, ornithine and citrulline supplementation upon performance and metabolism of trained rats. AB - During intense exercise there is an augmented production of ammonia and IMP in the exercised muscle that could be related to the establishment of peripheral fatigue. In order to prevent this accumulation, the urea cycle in the liver eliminates ammonia in the form of urea and the skeletal muscle buffers the increase of ammonia via transamination reactions. In the present study we evaluated the effect of arginine, citrulline and ornithine supplementation, intermediates of the urea cycle, on the performance of sedentary and swimming trained rats submitted to a single bout of exhaustive exercise. We also measured the glycogen content of the soleus and gastrocnemius muscles and of the liver, as well as the plasma concentrations of ammonia, urea, glutamine, glucose and lactate. The results indicate that arginine, citrulline and ornithine supplementation increased the flux of substrate through the reaction catalysed by glutamine synthetase, leading to increased glutamine production after an exhaustive bout of exercise, and of the mechanism involved in ammonia buffering. PMID- 12579528 TI - Determination of intracellular efficacies of azithromycin against Leishmania major infection in human neutrophils in vitro. AB - Azithromycin is one of a new class of antibiotics known as azalides. Azithromycin has high tissue affinity and this feature is thought to be due to the presence of two basic tertiary amine groups. Leishmania major, one of the causative agents of cutaneous leishmaniosis, is an obligate intracellular parasite. In this in vitro study, the potential anti-leishmanial effect of azithromycin upon intracellular forms namely the amastigote of L. major in mice peritoneal macrophages was investigated. L. major promastigotes were propagated in RPMI-1640 supplemented with 20% fetal calf serum in the log phase. The percentage of phagocytosis and microbiacidal activity of azithromycin on macrophages was assessed in the control and study groups by fluorescence microscopy, using acridine orange. Our results showed that at all the concentrations used (0.05, 0.1, 0.3, 0.6 microg ml(-1)) azithromycin had no inhibitory effect on the phagocytic capacity of mouse peritoneal macrophages. Although no significant difference was observed for leishmaniacidal activity between the study and the control groups at a concentration of 0.05 microg ml(-1) (p>0.05), a significant (p<0.05) increase in leishmaniacidal activity was detected at 0.1, 0.3 and 0.6 microg ml(-1). As a result, azithromycin does not provide any contribution to the phagocytosis of L. major promastigotes in macrophages in vitro, but it increases the intracellular killing rates of amastigotes. These results suggest that it has a potential anti leishmanial effect, and may provide a significant advantage in the treatment of the disease. PMID- 12579529 TI - Apoptosis and melanoma: molecular mechanisms. AB - Melanoma cells can undergo self-destruction via programmed cell death, i.e. apoptosis. In these tumours, the molecular components of apoptosis include positive (apoptotic) and negative (anti-apoptotic) regulators. The former include p53, Bid, Noxa, PUMA, Bax, TNF, TRAIL, Fas/FasL, PITSLRE, interferons, and c KIT/SCF. The latter include Bcl-2, Bcl-X(L), Mcl-1, NF-(K)B, survivin, livin, and ML-IAP. Alternatively, some molecules such as TRAF-2, c-Myc, endothelins, and integrins may have either pro- or anti-apoptotic effects. Some of these molecules are of potential therapeutic use, such as: (1) p53, which influences resistance to chemotherapy; (2) Mcl-1 and Bcl-X(L), which can override apoptosis; (3) TRAIL, which has selective fatal effects on tumour cells; (4) NF-(K)B, which when downregulated sensitizes cells to TRAIL and TNF; (5) the PITSLRE kinases, whose alteration appears to result in Fas resistance; (6) interferons, which sensitize cells to other factors; and (7) survivin and other IAPs that inhibit apoptosis. This review summarizes the state of current knowledge about the key molecular components and mechanisms of apoptosis in melanoma, discusses potential therapeutic ramifications, and provides directions for future research. PMID- 12579530 TI - Concomitant expression of p16INK4a and p14ARF in primary breast cancer and analysis of inactivation mechanisms. AB - The INK4a/ARF locus encodes two tumour suppressor proteins, p16INK4a and p14ARF, which act in the two main cell-cycle control pathways, p16-Rb and p14-p53 respectively. The present study examined the mRNA expression of these genes by reverse transcription-polymerase chain reaction (RT-PCR), and the inactivation mechanisms that alter these levels, in 100 primary breast carcinomas. Furthermore, the interdependence of these mechanisms was examined, since it has been reported that p14ARF is altered in most tumours in concordance with p16INK4a. The results show that promoter hypermethylation, tested by methylation specific PCR (MSP), was the major mechanism of inactivation of these genes and was present in 31 (31%) and 50 (50%) of the tumours that showed decreased p16INK4a and p14ARF expression, respectively. Hemizygous deletion was the second cause of down-regulation. Homozygous deletion was rare and mutation was absent. In most tumours overexpressing p16INK4a or p14ARF, no detectable inactivation mechanisms were observed. Finally, the results indicate that these proteins are often co-altered in primary breast tumours and that p16INK4a and p14ARF had non independent behaviour, since they were silenced or overexpressed concomitantly with a significant correlation (p < 0.05). PMID- 12579531 TI - Loss of types XV and XIX collagen precedes basement membrane invasion in ductal carcinoma of the female breast. AB - Ductal and lobular carcinomas comprise most malignancies of the female breast and the morbidity and mortality associated with breast cancer. During the progression from in situ to invasive stages, tumour cells penetrate the epithelial and vascular basement membranes (BM) to realize full metastatic potential. While the definition of these structures has primarily resulted from analysis of laminin and type IV collagen, characterization of newly discovered BM/BM zone (BMZ) proteins will further elucidate the interactions between tumour cells and the host stroma. We have studied the expression of two non-fibrillar BMZ collagens, the type XV proteoglycan and collagen XIX, in breast cancer where a linear, well formed BM becomes fragmented and even lost in the progression of epithelial malignancy. In the normal breast, types XV and XIX were found in all BMZ: epithelial, muscle, neural, endothelial, and fat. In in situ lesions, these two collagens, and particularly type XV, were often absent from the BM/BMZ displaying a continuous or just focally disrupted type IV/laminin staining pattern. In contrast, infiltrating ductal carcinomas showed only rare traces of laminin and collagen IV reactivity adjacent to the glands and tumour nests, and similarly there was little if any evidence of types XV and XIX collagen. All four molecules were, however, detected in the interstitium associated with some of the invasive carcinomas. The data suggest that types XV and XIX collagen are lost early in the development of invasive tumours, prior to penetration and eventual dissolution of the epithelial BM. Disappearance of these proteins from the BM/BMZ may signal remodelling of the extracellular matrix to promote tumour cell infiltration. PMID- 12579532 TI - Fluorescence lifetime imaging of unstained tissues: early results in human breast cancer. AB - Fluorescence lifetime imaging (FLIM) depends on the fluorescence decay differences between tissues to generate image contrast. In the present study FLIM has been applied to fixed (but unstained) breast cancer tissues to demonstrate the feasibility of this approach for histopathological assessment. As the FLIM method relies on natural autofluorescence, it may be possible to circumvent tissue processing altogether and so FLIM has the potential to be a powerful new method of in vivo tissue imaging via an endoscopic or per-operative approach in a variety of organs, as well as a research tool for in vivo animal models of disease. Unstained, alcohol-fixed tissue samples from 13 patients were stimulated by laser pulses at 415 nm. The temporal decay of the autofluorescence was imaged over a period of 2 ns after cessation of the pulse. The decay rate at each image pixel was calculated as the 'lifetime' factor tau. A tissue classification scheme was used to define regions in each image. The average lifetimes of different tissue regions were compared. A total of 167 tissue regions were measured. Within individual fields, stroma had a larger tau (slower decay) than epithelium (p < 0.001). Within individual patients (taking the mean tau of a given tissue type across all fields from each patient), there was a statistically significant difference between benign and malignancy-associated stroma (p < 0.05). Also, benign collagen had a longer tau than benign epithelium (p < 0.05). Multivariate analysis showed a significant difference between benign stroma, malignancy associated stroma, blood vessels, and malignant epithelium (p < 0.05). Statistically significant differences between benign and malignancy-associated stroma were obtained even with small patient numbers, indicating that lifetime based instruments can be developed for real-time diagnostic imaging with microscopic resolution. PMID- 12579533 TI - Increased expression of IgE-dependent histamine-releasing factor in endometriotic implants. AB - A complex network of cytokines mediates the immunoregulatory responses leading to endometriosis. Recent intensive studies suggest that monocyte and T cell chemoattractants contribute to the inflammatory environment of endometriotic implants. The relationship between the inflammation present during endometriosis and the development of endometriotic implants in the peritoneal cavity remains unclear. On the other hand, the association between endometriosis and 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD; dioxin) exposure has been discussed in recent years, and our previous results revealed that IgE-dependent histamine-releasing factor (HRF) is inducible by TCDD. The present study aimed to clarify the expression, localization, and function of HRF in endometriosis. Northern blot analysis demonstrated that HRF is overexpressed in endometriotic implants. RT-PCR with Southern blot analysis, however, showed that HRF overexpression was not always accompanied by CYP1A1 induction in endometriotic implants, suggesting that HRF is inducible in endometriosis without exposure to TCDD. HRF is also inducible by macrophage colony-stimulating factor (M-CSF). Immunohistochemistry showed CD68 positive macrophages in the stroma of endometriotic implants, adjacent to regions with prominent HRF accumulation. HRF-overexpressing cells exhibited high implantation efficiency in comparison to control cells when the cells were injected into the peritoneal cavities of nude mice. These results suggest that the accumulation of macrophages in endometriotic implants induces HRF; the overexpression of HRF accelerates the growth of endometriotic implants. PMID- 12579534 TI - Expression of haem oxygenase in cirrhotic rat liver. AB - The haem oxygenase (HO)/carbon monoxide (CO) system has been implicated as a modulator of hepatobiliary function. This study investigated HO expression in the process of cirrhosis development, as well as its relationship to nitric oxide synthase (NOS). Liver cirrhosis was induced in rats by chronic bile duct ligation (BDL). HO mRNA expression was evaluated by competitive RT-PCR, while protein expression was determined by immunoblotting and immunohistochemistry. In liver tissue where cirrhosis had fully developed, the expression levels of HO-1 were greatly enhanced at both mRNA and protein level compared with sham livers. Immunohistochemistry showed that HO-1 was induced in hepatocytes and enhanced in some of the Kupffer-like cells in BDL livers. In contrast, there was no difference between the sham and the BDL livers for the expression levels of HO-2. Interestingly, administration of the NOS inhibitor aminoguanidine (AG) or N(G) nitro-L-arginine methyl ester inhibited HO-1 expression. To study further the role of HO-1 in the development of liver cirrhosis, hepatocytes were isolated from the rats at different time points after BDL operation. HO-1 was expressed in hepatocytes at high levels during the early onset of cirrhosis but dropped slightly at a later stage of cirrhosis. Zinc protoporphyrin IX (ZnPP), an HO inhibitor, blocked HO-1 expression in hepatocytes from BDL cirrhotic rats, but enhanced the activity of inducible NOS (iNOS) in BDL hepatocytes. In conclusion, HO-1 was induced in the hepatocytes of rats undergoing cirrhosis, suggesting that HO-1 plays a role in the development of liver cirrhosis. Induction of HO-1 may be mediated partially by iNOS. However, once it is induced, HO-1 may be important in modulating iNOS activity, thus playing a protective role in liver cirrhosis. PMID- 12579535 TI - Early up-regulation of chemokine expression in fulminant hepatic failure. AB - CC-chemokines recruit and activate macrophages and T lymphocytes, the major components of inflammatory infiltrates in fulminant hepatic failure (FHF). To analyse the role of CC-chemokines in the pathogenesis of FHF, this study examined serum levels and intrahepatic expression of MCP-1, MIP-1alpha, MIP-1beta, and RANTES in the livers and sera of patients with FHF and controls by ELISA, immunohistochemistry, and competitive RT-PCR. Serum levels and intrahepatic expression of all chemokines studied in FHF exceeded the levels in chronic liver diseases and normal controls. Distinct patterns of expression of each chemokine were noted on Kupffer cells, sinusoidal endothelial cells, hepatocytes, lymphocytes, and bile ducts. Intrahepatic chemokine expression correlated closely with the extent of infiltration by macrophages and T lymphocytes (r = 0.65-0.95, p < 0.001). The functional relationship between intrahepatic chemokine release and infiltration was confirmed in chemotaxis assays by inhibiting chemotaxis induced by homogenates of liver tissue obtained from FHF patients with neutralizing MCP-1, MIP-1alpha, MIP-1beta, and RANTES antibodies. The time course of CC-chemokine release was studied in the concanavalin A and the galactosamine/LPS mouse models of FHF. In both models, intrahepatic chemokine up regulation occurred as an early event prior to hepatic infiltration and liver damage. The data indicate that an abundant intrahepatic release of CC-chemokines is an early and pivotal step in the pathogenesis of FHF. PMID- 12579536 TI - Determination of the molecular relationship between multiple tumour nodules in hepatocellular carcinoma differentiates multicentric origin from intrahepatic metastasis. AB - The purpose of this study was to determine the molecular relationship between multiple tumour nodules in hepatocellular carcinoma (HCC) within individual patients and to evaluate their clonality, which may bear prognostic significance. In 25 HCC nodules from 11 patients with multiple HCCs, the clonal relationships of the nodules within individual patients were determined using DNA fingerprinting with loss of heterozygosity (LOH) assay, comparative genomic hybridization (CGH), and hepatitis B virus (HBV) integration pattern. Both LOH assay and CGH indicated that in four (36%) of the 11 patients, the multiple HCCs had different clonalities and hence were of multicentric origin, whereas in the remaining seven (64%) patients, the multiple HCCs had similar clonal relationships and were intrahepatic metastases. In selected cases, the HBV integration pattern helped to confirm the clonality results. Gross appearance, size, location, and histology of the tumours could not accurately predict their clonal origin in every case. Assessment of DNA alterations allows precise determination of the clonality of multiple HCCs within one patient. Of these molecular methods, LOH analysis can be used to evaluate tumour clonality in most patients even before surgical resection, since this assay can be readily applied routinely to either liver biopsies or fine needle aspirates. PMID- 12579537 TI - Comparative molecular and histological grading of epithelial dysplasia of the oral cavity and the oropharynx. AB - Histological grading of epithelial dysplasia in the oral cavity and oropharynx is used to predict the risk for cancer and to determine the treatment strategy. This grading, however, is subjective and not well reproducible. Recent publications have shown that molecular markers are promising in cancer risk assessment. The aim of the present study was to compare classical histological and molecular grading and to relate these to the proliferation rate by quantitative assessment of Ki-67 staining. Forty-three samples were analysed from the margins of patients who had undergone resection of their squamous cell carcinoma in the oral cavity/oropharynx. Three experienced pathologists performed the histological grading. With the consensus score, 12 samples were classified as normal and 31 as dysplastic (21 mild, six moderate, and four severe). Loss of heterozygosity (LOH) was assessed in the same samples with 15 microsatellite markers at chromosomes 3p, 9p, 17p, 8p, 13q, and 18q, and was present in 28 of the 43 samples. Twenty four of the 28 cases (86%) with LOH were classified as dysplastic and four as normal. All ten samples with moderate and severe dysplasia and 14 of 21 samples with mild dysplasia contained LOH. In four of 12 biopsies classified as normal, LOH was found. A very striking and significant difference of the Ki-67 index was observed between LOH-positive and LOH-negative cases, 36.6 +/- 11.1% versus 19.4 +/- 2.8% positive cells, respectively. In mild dysplasia, 13 of 14 lesions containing LOH had a higher Ki-67 index than all seven lesions without LOH. Thus, in the oral cavity/oropharynx, LOH is more frequently found in the histologically higher-grade lesions (moderate dysplasia or worse) and in the lower grade lesions when a high proliferation rate is present. Assessment of proliferation with Ki-67 is a better surrogate for LOH than histological grading. PMID- 12579538 TI - Cross-reactivity of yeast antigens in human colon and peripheral leukocytes. AB - Elevation of the serum anti-Saccharomyces cerevisiae antibody (ASCA) level has been reported in patients with Crohn's disease. This study investigated the antigenic distribution of S. cerevisiae in human colon and peripheral leukocytes. ASCA was isolated from sera from patients with Crohn's disease using immuno affinity chromatography and then biotinylated and assayed immunohistologically and immunocytologically to determine the distribution of antigens recognized by ASCA in human colon and peripheral leukocytes. Immunoblot analysis of yeast extract and human peripheral leukocytes was performed. Immunohistological study using biotinylated ASCA revealed the presence of yeast-like particles in the granulation tissue of inflamed colonic mucosa. Biotinylated ASCA also stained lymphocytes and polymorphonuclear cells infiltrating inflamed intestine. Monocytes in epithelioid granulomas of colon with Crohn's disease were also stained. Polymorphonuclear leukocytes in peripheral blood were also stained with biotinylated ASCA. The antigens reactive to ASCA among heat-extracted, non-heat extracted yeast antigens, and human leukocyte extract differed. The findings of cross-reactivity of polymorphonuclear leukocytes with S. cerevisiae antigen and the presence of S. cerevisiae antigen in Crohn's disease granulomas suggest the possibility of involvement of S. cerevisiae in the pathogenesis of Crohn's disease. PMID- 12579539 TI - Markers for the development of early prostate cancer. AB - Biochemical and genetic changes precede histologically identifiable changes accompanying cell transformation often by months or years. De-expression of the extracellular matrix adhesive glycoprotein tenascin and the cell-to-cell adherent protein E-cadherin have been suggested as markers of early neoplastic change in prostate epithelial cells. Previous studies have been inconclusive, probably due to epitope masking. This study examined 2,378 biopsy cores from 289 prostates using a heat antigen retrieval protocol at low pH to improve the accuracy of detection. Tenascin and E-cadherin de-expression was correlated with purinergic receptor and telomerase-associated protein labelling, as well as prostate specific antigen (PSA) levels and Gleason scores. E-cadherin was a poor marker, as it was expressed in all lesions except carcinomas of the highest Gleason score. Tenascin was maximally expressed in the extracellular matrix and acinar basement membrane in normal and prostatic intraepithelial neoplasia tissue. In prostate cancer tissue, tenascin expression did not correlate with Gleason score but was significantly de-expressed as purinergic receptor and telomerase associated protein expression increased. Marked changes in tenascin, telomerase associated protein, and purinergic receptor expression were apparent before any histological abnormalities were visible by haematoxylin and eosin (H&E) stain, making these potential markers for early and developing prostate cancer. Moreover, the potential increased accuracy of diagnosis of underlying prostate cancer using purinergic receptor translocation (PRT) assessment suggests that PSA levels may be more accurate than has generally been supposed when apparent false negatives arising from H&E-based diagnoses are correctly categorized. PMID- 12579540 TI - Leydig cell micronodules are a common finding in testicular biopsies from men with impaired spermatogenesis and are associated with decreased testosterone/LH ratio. AB - To assess the biological significance of Leydig cell 'hyperplasia' in man, Leydig cell distribution, volume, and function were studied in patients with infertility or testicular cancer and in suddenly deceased controls. A total of 156 biopsies from 95 patients and 18 necropsies from 13 controls were examined using a semi quantitative stereological method. In patients, serum concentrations of testosterone, sex hormone binding globulin (SHBG), luteinizing hormone (LH), follicle stimulating hormone (FSH), oestradiol and inhibin-B were correlated with the findings on histological examination. Leydig cell clusters of more than 15 cells in a cross-section, for which we proposed the name 'micronodules', were frequently seen in testicles exhibiting Sertoli-cell-only syndrome (SCO), a mixed pattern of impaired spermatogenesis, or complete spermatogenesis in combination with elevated FSH. Median numbers of micronodules per 1.77 mm(2) (four fields of vision) in these three histological patterns were 6, 4, and 3.5, respectively. In contrast, micronodules were only occasionally observed in testicular biopsies from patients with complete spermatogenesis and normal gonadotrophin levels (median 1), and were rare in testes from controls (median = 0, p = 0.02). The proportion of testicular tissue occupied by Leydig cells increased with decreasing spermatogenic capacity. In contrast, the total volume of Leydig cells per testis was roughly comparable irrespective of the histological pattern, with the exception of testes with bilateral micronodules, which had significantly increased Leydig cell volume compared to those without micronodules. The number of micronodules correlated positively to LH (r = 0.577, p < 0.01) and FSH (r = 0.595, p < 0.01) and the presence of micronodules was most pronounced in the hyperstimulated testes, as reflected by an increased LH/testosterone ratio. In conclusion, Leydig cell micronodules were more frequent in biopsies with impaired spermatogenesis and associated with decreased ratios of testicular hormones to gonadotrophins. The presence of micronodules thus seems to be a histological marker of testicular failure in man. PMID- 12579541 TI - Alpha-actin isoform distribution in normal and failing human heart: a morphological, morphometric, and biochemical study. AB - We investigated the distribution of alpha-skeletal, alpha-cardiac, and alpha smooth muscle actin isoforms in human heart during development, hypertrophy, and failure. At 20 weeks of fetal life, alpha-skeletal actin was localized in a small proportion of subendocardial and papillary muscle cardiomyocytes. At this gestation time, diffuse alpha-cardiac actin staining was observed, associated with focal expression of alpha-smooth muscle actin. In normal adult subjects, alpha-skeletal actin positive cardiomyocytes were distributed in a transmural gradient with the highest proportion located subendocardially. In myocardial hypertrophy and cardiomyopathies, the amount of alpha-skeletal actin was increased and diffuse staining was seen in all layers of ventricular myocardium, with the exception of idiopathic dilated cardiomyopathies. Cardiomyocytes were negative for alpha-smooth muscle actin in all pathological situations studied. As expected, fibroblasts in post-infarct scars expressed alpha-smooth muscle actin and transforming growth factor-beta1 but, surprisingly, were negative for these proteins in interstitial fibrosis. Our results demonstrate that increased expression of alpha-skeletal actin in the diseased human heart is associated with increased myocyte stretch, increased wall stress, and pressure overload, but not with idiopathic dilated cardiomyopathies. They also suggest that fibrotic changes develop with different mechanisms in scars versus interstitial fibrosis. PMID- 12579542 TI - The pancreatitis-associated protein induces lung inflammation in the rat through activation of TNFalpha expression in hepatocytes. AB - The pancreatitis-associated protein (PAP) is a pancreatic stress protein overexpressed during acute pancreatitis, a disease often accompanied by lung inflammation. We investigated whether PAP was involved in the occurrence of this remote complication of pancreatitis and whether the liver might be implicated in the process. PAP was injected into the vena cava of rats (40 or 400 micro g/kg body weight). For comparison, pancreatitis was induced in rats by intraductal administration of sodium taurocholate. Three hours later, parameters of inflammation and mRNA concentrations of TNFalpha, P-selectin, heat shock protein (HSP)-70, and extracellular superoxide dismutase (EC-SOD) were monitored in lung and liver. Significant increases in P-selectin expression, neutrophil infiltration, and oxidative stress revealed that PAP treatment induced lung inflammation in rats and exacerbated inflammation in animals with pancreatitis. Plasma TNFalpha level was increased and TNFalpha mRNA was strongly overexpressed in liver, with concomitant activation of NF-kappaB; in situ hybridization revealed that TNFalpha overexpression was mainly located to hepatocytes. Lung inflammation induced by PAP could be prevented by injection of anti-TNFalpha antibodies. It was concluded that, during pancreatitis, PAP released by the pancreas could mediate lung inflammation through induction of hepatic TNFalpha expression and subsequent increase in circulating TNFalpha. PMID- 12579544 TI - Self-help and minimal-contact therapies for anxiety disorders: Is human contact necessary for therapeutic efficacy? AB - Self-help materials, brief therapies, and treatments involving minimal therapist contact have all been proposed as effective and low-cost interventions for anxiety disorders. However, research also suggests that the therapeutic alliance is a central predictor of therapy outcome. Interestingly, amounts of therapist contact within and across "self-help" interventions vary greatly. It is therefore unclear how much therapist contact is necessary for a positive anxiety disorder treatment outcome. The present article reviews the literature on anxiety disorder treatments using self-help, self-administered, and decreased therapist-contact interventions. Treatment studies are grouped together by anxiety diagnosis as well as amount of therapist contact. It is concluded that self-administered treatments are most effective for motivated clients seeking treatment for simple phobias. Predominantly self-help therapies are efficacious for panic disorder and mixed anxiety samples. On the other hand, minimal-contact therapies have demonstrated efficacy for the greatest variety of anxiety diagnoses. PMID- 12579545 TI - Self-administered treatments for depression: a review. AB - Although there are numerous self-help books for depression, relatively few have been empirically tested. However, those that have been used in clinical trials have fared well, with an average effect size roughly equivalent to the average effect size obtained in psychotherapy studies. Computer-based treatments are being developed and appear promising as an alternative to bibliotherapy for those interested in self-administered treatments. This article provides a summary of the depression bibliotherapy literature and discusses several remaining questions such as effectiveness versus efficacy, practice applications, ethics, and future research. PMID- 12579546 TI - A meta-analysis of the effectiveness of bibliotherapy for alcohol problems. AB - There has been increased interest in the use of brief interventions and the delivery of alcohol treatment services through nonspecialist health care settings. One possible resource for reaching untreated individuals is "bibliotherapy," the provision of self-help materials to motivate and guide the process of changing drinking behavior. Research on the effectiveness of self-help materials for problem drinkers has been done for three decades. This report summarizes a meta-analytic review of 22 studies evaluating the effectiveness of such self-help materials. Each study was rated on 12 methodological criteria, and effect sizes of bibliotherapy were computed. The methodological quality of studies was generally high relative to other treatment-outcome studies. Modest support was found for the efficacy of self-help materials in decreasing at-risk and harmful drinking. The weighted mean pre/post-effect size for bibliotherapy was .80 with self-referred individuals seeking help for drinking problems, and .65 for individuals identified through health screening. Between-group comparisons of bibliotherapy with no-intervention controls appear to have a small to medium effect, with a weighted mean effect size of .31 with self-referred drinkers; effect size was more variable in opportunistic interventions based on health screening. Finally, between-group comparisons of effects on drinking of bibliotherapy versus more extensive interventions yielded effect size values near zero. These findings provide support for the cost-effective use of bibliotherapy with problem drinkers seeking such help to reduce their consumption, and to a lesser extent with drinkers who are identified through screening as at risk. PMID- 12579547 TI - Self-administered treatment for smoking cessation. AB - Self-administered treatment for smoking cessation has the potential to reach a broad spectrum of the population of smokers. This article focuses on self administration of behavioral and pharmacological treatments for smoking cessation. Evidence for the effectiveness of written manuals to self-administer behavioral treatment is mixed. There is no evidence that self-help manuals alone are effective. However, they do increase quit rates when combined with personalized adjuncts such as written feedback and outreach telephone counseling. Efficacy trials of first-line pharmacotherapies (nicotine gum, nicotine patch, and bupropion) result in doubling of cessation rates compared to placebo. It is difficult to evaluate the effectiveness of pharmacotherapies when self administered under real-world conditions. The general consensus is that they improve quit rates, although poor compliance and early discontinuation reduce their effectiveness. Areas for further research include randomized trials of the use of new technologies (e.g., hand-held computers and the Internet) to disseminate self-administered treatments as well as improved surveillance of the use of self-administered treatment in population-based health surveys. PMID- 12579549 TI - Self-administered treatment in stepped-care models of depression treatment. AB - Stepped behavioral health care models have begun to receive increased attention. Self-administered treatments deserve consideration as an element in these models for some disorders and for some consumers. Features suggesting inclusion include low cost, wide availability, and evidence-based status. We present a stepped-care model for depression inclusive of a self-administered treatment component. We also discuss cautions such as depression severity and consumer preference. Evaluation of the efficacy and cost effectiveness of this approach to depression treatment is necessary. PMID- 12579548 TI - Self-administered psychosocial treatments for children and families. AB - Self-administered psychosocial treatments for child health problems have the potential to circumvent barriers to traditional models of care. They are convenient and inexpensive to families and, for some types of problems, may be as effective as therapist-based care. In a review of past research on child- and parent-facilitated self-administered treatments, it was found that a stronger evidence base exists in support of some formats (manual- and multimedia-based treatments) than for others (inspirational literature and support groups). The practical, ethical, and legal issues associated with self-administered treatments are discussed as well as avenues for future research. How psychologists and health care systems respond to the opportunities associated with self administered treatments for children will likely affect the face of the profession and the health of children in the future. PMID- 12579550 TI - Development of a Controlled Drinking Self-Efficacy Scale and appraising its relation to alcohol dependence. AB - There is no specific self-efficacy measure that has been developed primarily for problem drinkers seeking a moderation drinking goal. In this article, we report the factor structure of a 20-item Controlled Drinking Self-Efficacy Scale (CDSES; Sitharthan et al., 1996; Sitharthan et al., 1997). The results indicate that the CDSES is highly reliable, and the factor analysis using the full sample identified four factors: negative affect, positive mood/social context, frequency of drinking, and consumption quantity. A similar factor structure was obtained for the subsample of men. In contrast, only three factors emerged in the analysis of data on female participants. Compared to women, men had low self-efficacy to control their drinking in situations relating to positive mood/social context, and subjects with high alcohol dependence had low self-efficacy for situations relating to negative affect, social situations, and drinking less frequently. The CDSES can be a useful measure in treatment programs providing a moderation drinking goal. PMID- 12579551 TI - Who plays the client? Collaborating with theater departments to enhance clinical psychology role-play training exercises. AB - This article offers a description of a student-active method of teaching clinical psychology applications, such as interviewing and psychotherapy, that involves collaboration between psychology and theater departments. Clinical psychology instructors recruit theater students to play the client roles in live dyads with the instructors or other students. These dyads may take place during or outside of class and may be videotaped for use in other educational settings. Practical and logistical issues are discussed, and empirical data are presented to demonstrate the effectiveness of this method. PMID- 12579552 TI - The role of activation in the effect of total sleep deprivation on depressed mood. AB - The present investigation analyzes if the effect of Total Sleep Deprivation (TSD) on depressive mood in normal subjects is related to participants' activation levels. Reaction time (RT), subjective sleepiness, and depressive mood were assessed among 48 normal subjects who were subjected to 60 h of TSD. Subjects were divided into groups based on his/her depressive mood changes following TSD: subjects with worse response (n = 11), subjects with better response (n = 12), and intermediate group (n = 25). We sought to determine if RT and subjective sleepiness were related systematically to depressive mood changes following TSD. TSD produces a general reduction in activation (i.e., a monotonic increase in RT and subjective sleepiness) modulated by circadian oscillations. Baseline activation (RT) was confirmed partially as a predictor of participants' response to TSD. Subjects with high levels of activation (lower RTs at the baseline) experienced a better response (greater reductions in depressive mood) following TSD. There was no relationship between pre-treatment subjective sleepiness levels and the TSD effect on depressive mood. The results partially support the role of the activation level as a differential moderator of TSD effects on depressive mood in normal subjects. PMID- 12579553 TI - Clinical presentations in combat veterans diagnosed with posttraumatic stress disorder. AB - This article investigated subtypes of symptom patterns among male combat veterans diagnosed with posttraumatic stress disorder (PTSD) through a cluster analysis of their Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Graham, Ben Porath, Tellegen, Dahlstrom, & Kaemmer, 2001) clinical and validity scales. Participants were 126 veterans seeking outpatient treatment for combat-related PTSD at a Veterans Affairs Medical Center. Two well-fitting MMPI-2 cluster solutions (a four-cluster solution and a three-cluster solution) were evaluated with several statistical methods. A four-cluster solution was determined to best fit the data. Follow-up analyses demonstrated between-cluster differences on MMPI 2 "fake bad" scales and content scales, the Beck Depression Inventory (BDI; Beck, Ward, Mendelson, Mock, & Erbaugh, 1961), Dissociative Experiences Scale (DES; Bernstein & Putnam, 1986), Mississippi Combat PTSD scale (M-PTSD; Keane, Caddall, & Taylor, 1988), and Clinician-Administered PTSD Scale (CAPS-1; Blake et al., 1990). Clusters also were different in disability-seeking status, employment status, and income. Implications for research and clinical practice using the MMPI-2 with combat veterans presenting with PTSD are briefly addressed. PMID- 12579554 TI - Suppression of the inflammatory response from adherent cells on phospholipid polymers. AB - The expression of interleukin-1beta (IL-1beta) messenger RNA (mRNA) in macrophage like cells cultured on phospholipid polymers was evaluated to determine the extent of the inflammatory response. As phospholipid polymers, poly(2 methacryloyloxyethyl phosphorylcholine(MPC)-co-n-butyl methacrylate(BMA)s (PMBs) were synthesized. Poly(ethylene terephthalate) (PET), poly(2-hydroxyethyl methacrylate) (PHEMA), and segmented poly(ether urethane) (Tecoflex 60) were used as reference biomedical polymers. The protein adsorption onto the polymer surfaces from a cell culture medium was determined. The amount of the total protein adsorbed onto the PMBs was lower than that adsorbed onto the reference polymers, and the amount of adsorbed protein decreased with an increase in the MPC units in the PMBs. Human premyelocytic leukemia cell line (HL-60) was used, and the expression of IL-1beta mRNA was investigated with the reverse transcription polymerase chain reaction (RT-PCR) method. When HL-60 cells were cultured on PMBs, the expression of IL-1beta mRNA in the cells was much less than that on the reference polymers. In particular, the expression of IL-1beta mRNA in HL-60 cells cultured on the PMBs containing more than 10 mol % MPC units was not detected. This corresponded to the reduced amount of adsorbed proteins on the PMB surfaces. These results suggest that the PMBs effectively suppressed the activation and inflammatory response of adherent macrophagelike cells. PMID- 12579555 TI - Bone formation in CaP-coated and noncoated titanium fiber mesh. AB - The osteogenic activity of calcium phosphate (CaP)-coated and noncoated porous titanium (Ti) fiber mesh loaded with cultured syngeneic osteogenic cells after prolonged in situ culturing was compared in a syngeneic rat ectopic assay model. Rat bone marrow (RBM) cells were loaded onto the CaP-coated and noncoated Ti scaffolds using either a droplet or a suspension loading method. After loading, the RBM cells were cultured for 8 days in vitro. Thereafter, implants were subcutaneously placed in 39 syngeneic rats. The rats were euthanized and the implants retrieved at 2, 4, and 8 weeks postoperatively. Further, in the 8 week group fluorochrome bone markers were injected at 2, 4, and 6 weeks. Histological analysis demonstrated that only the CaP-coated meshes supported bone formation. The amount of newly formed bone varied between single and multiple spheres to filling a significant part of the mesh porosity. In the newly formed bone, osteocytes embedded in a mineralized matrix could be observed clearly. On the other hand, in the noncoated titanium implants, abundant deposition of calcium containing material was seen. This deposit lacked a bonelike tissue organization. Further analysis revealed that the cell-loading method did not influence the final amount of bone formation. In CaP-coated implants the accumulation sequence of the fluorochrome markers showed that bone formation started on the mesh fibers. In conclusion, our results prove that the combination of a thin CaP coating, Ti-mesh, and RBM cells can indeed generate ectopic bone formation after prolonged in vitro culturing. No effect of the loading method was observed on the final amount of bone. PMID- 12579556 TI - Crosslinking of biological tissues using genipin and/or carbodiimide. AB - The study was to investigate the crosslinking characteristics, mechanical properties, and resistance against enzymatic degradation of biological tissues after fixation with genipin (a naturally occurring crosslinking agent) and/or carbodiimide. Fresh tissue was used as a control. It was found that both genipin and carbodiimide are effective crosslinking agents for tissue fixation and genipin crosslinking is comparatively slower than carbodiimide crosslinking. Additionally, tissue fixation in genipin and/or carbodiimide may produce distinct crosslinking structures. Carbodiimide may form intrahelical and interhelical crosslinks within or between tropocollagen molecules, whereas genipin may further introduce intermicrofibrillar crosslinks between adjacent collagen microfibrils. The stability (denaturation temperature and resistance against enzymatic degradation) of the fixed tissue is mainly determined by its intrahelical and interhelical crosslinks. In contrast, intermicrofibrillar crosslinks significantly affect the mechanical properties (tissue shrinkage during fixation, tensile strength, strain at break, and ruptured pattern) of the fixed tissue. Moreover, the degree of enzymatic degradation of the fixed tissue may be influenced by three factors: the availability, to the enzyme, of recognizable cleavage sites, the degree of crosslinking, and the extent of helical integrity of tropocollagen molecules in tissue. PMID- 12579557 TI - Neural differentiation of midbrain cells on various protein-immobilized polyethylene films. AB - The effect of surface modification of polyethylene (PE) film on differentiation of midbrain (MB) cells obtained from rat embryos was determined by their micromass culture system. When cultured on untreated PE film, cell differentiation was suppressed to approximately two-thirds of that observed in a control culture dish. On the contrary, type I collagen-immobilized PE film increased differentiated foci of the MB cells more than did the untreated PE film. RGDS (Arg-Gly-Asp-Ser) peptide immobilization onto PE film resulted in almost the same differentiation activity as the collagen immobilized PE film. Bovine serum albumin (BSA) immobilization onto PE film enhance the differentiation activity more than did the untreated PE film, but not up to the levels of collagen- and RGDS-immobilized PE. The number of differentiated foci of the MB cells on untreated PE film were increased by the addition of the condition medium prepared from the collagen-immobilized PE film. However, the number of foci was not increased by the addition of other condition media obtained from control dish, untreated, BSA-, and RGDS-immobilized PE. On the other hand, none of these condition media enhanced a differentiation of the neuronal cell line of PC12 cells, suggesting that some factors effectively differentiate midbrain cells, composed of neuronal epithelial and mesenchymal cells, but not the PC12 cells secreted in the condition media prepared from collagen-immobilized PE. In addition, it is probable that neural growth factor was not secreted in these condition media, which could not induce the differentiation of PC12 cells. PMID- 12579558 TI - Water-borne, in situ crosslinked biomaterials from phase-segregated precursors. AB - A novel process for the preparation of water-borne biomaterials for hard tissue repair from injectable precursors is described, where the precursors form crosslinked materials in situ under physiological conditions. The precursors react by means of a Michael-type addition reaction that makes use of addition donors such as pentaerythritol tetrakis 3'-mercaptopropionate (QT) and addition acceptors such as poly(ethylene glycol) diacrylate 570 MW (PEGDA), pentaerythritol triacrylate (TA), and poly(propylene glycol) diacrylate 900 MW (PPODA). These crosslinked materials (at 75 wt% solid), prepared from water dispersions or reverse emulsions, showed ultimate strengths in compression of 1.8 +/- 0.2 and 6.7 +/- 0.5 MPa and ultimate deformations of 35 +/- 2+/- and 37 +/- 2%, respectively. Scanning electron microscopy (SEM) shows that the morphology of the precursors templated the morphology of the final materials. The current study indicates that it is possible to obtain injectable high-modulus materials that have appropriate mechanical properties and gelation kinetics for tissue augmentation and stabilization applications such as mechanical stabilization of the intervertebral disc annulus. PMID- 12579559 TI - Morphological characteristics of total joint arthroplasty-derived ultra-high molecular weight polyethylene (UHMWPE) wear debris that provoke inflammation in a murine model of inflammation. AB - It is recognized that the chronic inflammation in peri-prosthetic tissue that contributes to implant failure frequently is provoked by the presence of wear debris. Some wear debris is inevitable because of the nature of the prosthesis, but not all patients develop severe inflammatory responses. The precise factors that mediate the severity of tissue inflammation to wear debris has yet to be fully defined. Because wear debris retrieved from peri-prosthetic tissue consists of a heterogeneous mixture of materials with various sizes and shapes, this study evaluated the influence of two major physical aspects of ultra-high molecular weight polyethylene (UHMWPE) wear debris (shape and surface texture) using a model of tissue inflammation. UHMWPE debris particulates recovered from 50 peri prosthetic tissue samples were examined by scanning electron microscopy and categorized into four groups based upon aspect ratio and surface texture of the material. The four groups were defined as: 1) smooth and globular, 2) smooth and fibular, 3) rough and globular, and 4) rough and fibular. Histological analysis and ELISA assays were conducted to evaluate variations in cellular responses and cytokine production between the groups. The strongest expression of tumor necrosis factor alpha and interleukin-1 beta was found in tissues exposed to UHMWPE debris with both a rough surface texture and fibular shape, and this response was significantly elevated over debris particles with a smooth surface texture and globular shape. The data suggest that both shape and texture influence the severity of specific inflammatory responses and that rough debris surface texture exerts a marked effect on adverse tissue responses when combined with particles that have a sharp, elongated shape. PMID- 12579560 TI - Three-dimensional, bioactive, biodegradable, polymer-bioactive glass composite scaffolds with improved mechanical properties support collagen synthesis and mineralization of human osteoblast-like cells in vitro. AB - In the past decade, tissue engineering-based bone grafting has emerged as a viable alternative to biological and synthetic grafts. The biomaterial component is a critical determinant of the ultimate success of the tissue-engineered graft. Because no single existing material possesses all the necessary properties required in an ideal bone graft, our approach has been to develop a three dimensional (3-D), porous composite of polylactide-co-glycolide (PLAGA) and 45S5 bioactive glass (BG) that is biodegradable, bioactive, and suitable as a scaffold for bone tissue engineering (PLAGA-BG composite). The objectives of this study were to examine the mechanical properties of a PLAGA-BG matrix, to evaluate the response of human osteoblast-like cells to the PLAGA-BG composite, and to evaluate the ability of the composite to form a surface calcium phosphate layer in vitro. Structural and mechanical properties of PLAGA-BG were measured, and the formation of a surface calcium phosphate layer was evaluated by surface analysis methods. The growth and differentiation of human osteoblast-like cells on PLAGA BG were also examined. A hypothesis was that the combination of PLAGA with BG would result in a biocompatible and bioactive composite, capable of supporting osteoblast adhesion, growth and differentiation, with mechanical properties superior to PLAGA alone. The addition of bioactive glass granules to the PLAGA matrix resulted in a structure with higher compressive modulus than PLAGA alone. Moreover, the PLAGA-BA composite was found to be a bioactive material, as it formed surface calcium phosphate deposits in a simulated body fluid (SBF), and in the presence of cells and serum proteins. The composite supported osteoblast-like morphology, stained positively for alkaline phosphatase, and supported higher levels of Type I collagen synthesis than tissue culture polystyrene controls. We have successfully developed a degradable, porous, polymer bioactive glass composite possessing improved mechanical properties and osteointegrative potential compared to degradable polymers of poly(lactic acid-glycolic acid) alone. Future work will focus on the optimization of the composite scaffold for bone tissue-engineering applications and the evaluation of the 3-D composite in an in vivo model. PMID- 12579561 TI - Mixed-ligand modification of polyamidoamine dendrimers to develop an effective scaffold for maintenance of hepatocyte spheroids. AB - Compared with a monolayer culture, hepatocyte spheroids are known to maintain liver function for long periods. We found that hepatocytes formed spheroids when cultured on polyamidoamine dendrimers modified with fructose. Because galactose is a ligand for the asialoglycoprotein receptor on the hepatocyte cytoplasmic membrane, it was chosen as another ligand for modification in order to maintain adhesion of spheroids for long periods. Simultaneous modification of dendrimers with fructose and galactose had a marked effect on the time length of spheroid adhesion. Suppression of apoptosis and necrosis was observed in hepatocyte spheroids cultured on a dendrimer modified with fructose and galactose (F/G dendrimer). Moreover, the hepatocyte spheroids cultured on the F/G dendrimer had higher activities of liver-specific functions, such as urea synthesis and albumin gene expression, than did those cultured on single-ligand-modified dendrimers. The expression of heat shock protein (HSP) genes was examined to evaluate the stress response of cells to scaffolds. The hepatocytes cultured on the F/G dendrimer had very low expression levels of both HSP60 and HSP70 mRNAs. Thus immobilization of mixed-ligand-modified dendrimers could generate a suitable surface for hepatocyte spheroid formation. These dendrimers could be a powerful tool for generating custom-made scaffolds for cells other than hepatocytes by selecting the ligands suitable for each cell type. PMID- 12579562 TI - Modification of calcium-phosphate coatings on titanium by recombinant amelogenin. AB - Amelogenin proteins, the principal components of the developing dental enamel extracellular matrix, have been postulated to facilitate the elongated and oriented growth of the carbonated apatite crystals during enamel formation. We previously reported that amelogenin caused modulation of apatite crystals nucleated on a bioactive glass (Bioglass(R)) in vitro. Here, the effects of amelogenin on the growth morphology of calcium-phosphate crystals nucleated on a titanium surface were investigated in order to gain a better understanding of the role of amelogenins during enamel biomineralization and to explore their potential application in the design and development of novel biomaterials. The dose-dependent effects of a recombinant mouse amelogenin (rM179) were found to be different from those of bovine serum albumin, which significantly inhibited apatite crystal growth and caused the octacalcium phosphate (OCP) crystals to change from a plate-like shape to a curved shape, indicating a general inhibitory effect. The effects of rM179 on the crystal growth of OCP at 12.5-100 microg/mL and of apatite at 50 microg/mL were insignificant while the apatite crystals were remarkably elongated along their c-axes upon the use of 100 microg/mL of rM179. The unique modulation of the calcium-phosphate coatings on titanium by rM179 supports the view that amelogenins have a great potential for applications designed to develop novel biomimetic materials. PMID- 12579563 TI - Tissue response to a newly developed calcium phosphate cement containing succinic acid and carboxymethyl-chitin. AB - We developed a new calcium phosphate cement containing succinic acid and carboxymethyl-chitin in the liquid component. In this study, the biocompatibility and osteoconductivity of this new cement were investigated. After mixing, cement in putty form was implanted immediately between the periosteum and parietal bone and in the subcutaneous tissues of rats. In control cement, distilled water was used instead of the liquid component. In addition to histological evaluations, analyses with X-ray diffraction and Fourier transform infrared were performed for the subcutaneously implanted cements. Histological examination showed slight inflammation around the new cement on the bone and in the subcutaneous tissue at 1 week after surgery. At 2 weeks, the cement was partially bound to the parietal bone. The extent of the surface of the new cement directly in contact with the bone increased with time, and most of the undersurface of the new cement bound to the host parietal bone by 8 weeks. Analysis by X-ray diffraction showed that the new cement in the subcutaneous tissue was transformed into hydroxyapatite by 8 weeks. These results indicate that this new calcium phosphate cement is useful as a bone substitute material. PMID- 12579564 TI - Distribution patterns of the membrane glycoprotein CD44 during the foreign-body reaction to a degradable biomaterial in rats and mice. AB - Although biomaterials have been used in the clinical setting for a long time, little is known of the molecular mechanisms underlying the foreign-body reaction (FBR). A good understanding of these mechanisms is requisite for the controlled regulation of the FBR needed to prevent adverse tissue reactions and thus to improve the function of the biomaterial. Macrophages are essential in the inflammatory reaction in, as well as around, the implants, and they also are believed to initiate most of the adverse responses. Typically, during the FBR macrophages become activated and fuse into multinucleated giant cells (MnGCs). CD44, an integral membrane glycoprotein expressed on a broad spectrum of cell types, is involved in MnGC formation in vitro and in inflammation processes in general. In vivo it is not known whether CD44 is part of a specific protein machinery that enables macrophage fusion or whether it has additional functions in the FBR. In the present in vivo study, CD44 expression patterns were followed in rats and mice during the FBR to a degradable collagen type I biomaterial. We found that CD44 is upregulated on all migrating cells and on newly formed blood vessels at the onset of the FBR and that MnGCs, up to week 15 postimplantation, expressed CD44. Although no evidence was found that CD44 participates in macrophage fusion leading to multinucleation, it nevertheless may be an interesting target molecule for modulating the FBR in vivo, possibly by affecting cell activation, cell migration towards the biomaterial, vascularization, and MnGC formation. PMID- 12579565 TI - Bond strength, compositional, and structural properties of hydroxyapatite coating on Ti, ZrO2-coated Ti, and TPS-coated Ti substrate. AB - The effect of titanium plasma-sprayed (TPS) and zirconia (ZrO(2))-coated titanium (Ti) substrates on the adhesive, compositional, and structural properties of plasma-sprayed hydroxyapatite (HA) coatings were evaluated. X-ray diffraction, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy, energy dispersive spectroscopy, surface roughness, and adhesive strength were used to characterize the coatings. Apatite-type and alpha-tricalcium phosphate phases were observed for all HA coatings. A structural change due to the absence of a 960 cm(-1) peak during FTIR analysis was observed for all HA coatings. The coating surfaces appeared rough and melted, with surface roughness correlating to the size of the starting powder. No significant difference in the Ca/P ratio of HA on Ti and TPS-coated Ti substrates was observed. However, the Ca/P ratio of HA on ZrO(2)-coated Ti substrate was significantly increased. Interfaces between all coatings and substrates were observed to be dense and tightly bound, except for HA coatings on TPS-coated Ti substrate interface. However, an intermediate TPS or ZrO(2) layer between the HA and Ti substrate resulted in a lower adhesive strength as compared to HA on Ti substrate. PMID- 12579566 TI - Cell-based bonding of articular cartilage: An extended study. AB - This study evaluated the biomechanical characteristics of newly formed cartilaginous tissue synthesized from isolated chondrocytes and seeded onto devitalized cartilage in an extended study in vivo. Cartilage from porcine articular joints was cut into regular discs and devitalized by multiple freeze thaw cycles. Articular chondrocytes were enzymatically isolated and incubated in suspension culture in the presence of devitalized cartilage discs for 21 days. This procedure allowed the isolated chondrocytes to adhere to the devitalized matrix surfaces. Chondrocyte-matrix constructs were assembled with fibrin glue and implanted in dorsal subcutaneous pockets in nude mice for up to 8 months. Histological evaluation and biomechanical testing were performed to quantify the integration of cartilage pieces and the mechanical properties of the constructs over time. Histological analysis indicated that chondrocytes grown on devitalized cartilage discs produced new matrix that bonded and integrated individual cartilage elements with mechanically functional tissue. Biomechanical testing demonstrated a time dependent increase in tensile strength, failure strain, failure energy, and tensile modulus to values 5-30% of normal articular cartilage by 8 months in vivo. The values recorded at 4 months were not statistically different from those collected at the latest time point, indicating that the limits of the biomechanical property values were reached after four months from implantation. PMID- 12579567 TI - RGTA modulates the healing pattern of a defect in a monolayer of osteoblastic cells by acting on both proliferation and migration. AB - A family of heparan-like polymers, RGTAs, was shown to promote repair of various tissues. Like heparin and heparan-sulfates, RGTAs potentiate in vitro the biological activities of heparin-binding growth factors (HBGFs) and protect them against proteolytic degradation. It was postulated that RGTAs stimulate bone healing by interacting with HBGFs released in the wound site and, subsequently, by promoting the proliferation of cells implicated in this process. In a previous report, we examined how RGTA can modulate the proliferation of MC3T3-E1 osteoblastic cells. To further complete this study and to support this hypothesis, we developed an in vitro model of bone repair and examined the effects of RGTA alone or in association with FGF2, BMP-2, and TGF-beta1 which are representative of HBGFs known to stimulate bone repair. The model consisted of a 6-mm reproducible defect created on a MC3T3-E1 cell monolayer. In the presence of the different products added to the medium, the process of wound repair was measured through the filling of the acellular defect. We show that in 8 days, RGTA slightly inhibits repair alone compared to the control (2% FBS), that it inhibits the mitogenic effect of FGF2, and that it amplifies the inhibitory effect of BMP-2 and TGF-beta1. Repair was realized by an association of cell migration and cell proliferation mechanisms. To determine the part played by each process, DNA synthesis was evaluated for cell proliferation using an immunodetection technique [to measure incorporation of 5-bromo-2-deoxyuridine (BrdU)], coupled with a computer-assisted image analysis. The results show that the presence of RGTA (1) amplified the number of labeled nuclei compared to the control, (2) added to FGF2 or TGF-beta1, it reduced the number of labeled nuclei compared to FGF2 or TGF-beta1 alone, and (3) in the presence of BMP-2, it amplified the number of labeled nuclei compared to BMP-2 alone. Proper interpretation of these data requires a better understanding of the mechanism of action of RGTA on bone healing. PMID- 12579568 TI - Improvement of the biocompatibility of alginate/poly-L-lysine/alginate microcapsules by the use of epimerized alginate as a coating. AB - Alginate/poly-L-lysine(PLL)/alginate capsules are used widely for the microencapsulation of cells. Alginate consists of guluronic acid and mannuronic acid, the ratio and sequence of which affect the properties of the alginate. Using C5-epimerases, mannuronic acid can be converted to guluronic acid in the alginate polymer. Such an enzyme, AlgE4, was used to convert blocks of mannuronic acid (M-blocks) to blocks of alternating sequence (MG-blocks). The aims of this study were 1) to investigate whether the use of epimerized alginate as a coating could improve the biocompatibility of alginate/PLL/alginate capsules and 2) to study the biocompatibility of simple alginate beads prepared with epimerized alginate. Four different capsules, two of which contained epimerized alginate, were investigated after implantation in C57BL/6 mice for 1 week. The biocompatibility of alginate/PLL/alginate capsules, as measured by retrieval rates of the capsules and DNA contents and glucose oxidation rates of the cellular overgrowth, was improved when an epimerized coating alginate was used. There were, however, no statistically significant differences in the biocompatibility of simple alginate beads made from epimerized alginate when compared with non-epimerized alginate beads. In general, such beads produced without a PLL coating swelled to a higher extent than the conventional alginate/PLL/alginate capsules. In conclusion, the use of an epimerized coating on alginate-PLL-alginate can improve the biocompatibility of such capsules but still cannot completely eliminate the detrimental effects of PLL on the biocompatibility of the capsules. PMID- 12579569 TI - Microcapsules made by enzymatically tailored alginate. AB - Alginate is widely used for encapsulation of cells. Alginate is a linear block copolymer consisting of mannuronic acid (M) and guluronic acid (G). It has been shown that enzymes known as C-5 epimerases convert M to G in the polymer chain, giving rise to novel alginates with tailored properties. One of these enzymes, AlgE4, converts M blocks into blocks of strictly alternating M and G. In this study we investigated how alginate epimerized by AlgE4 affected capsule properties such as stability and permeability. Inhomogeneous calcium-alginate gel beads were made with original and AlgE4-epimerized alginates of different origin. The epimerized alginates formed initially smaller alginate gels that showed increased resistance to osmotic swelling compared with the original nonmodified alginate samples. The permeability, measured as diffusion of immunoglobulin (Ig) G into Ca/Ba-alginate gel beads, was reduced by epimerization and further reduced by addition of poly-L-lysine (PLL). The osmotic stability of alginate-poly-D lysine(PDL)-alginate capsules was enhanced by the use of epimerized alginate; indeed, stable capsules with low permeability to tumor necrosis factor (TNF) could be made with low PDL exposures. Finally, alginate with an alternating structure interacted more strongly with the alginate-PLL capsule than did alginate with a high content of M blocks or G blocks or than an alginate consisting mainly of M. PMID- 12579570 TI - Controlled release of fibroblast growth factors and heparin from photocrosslinked chitosan hydrogels and subsequent effect on in vivo vascularization. AB - Application of ultraviolet (UV) irradiation to a photocrosslinkable chitosan (Az CH-LA) aqueous solution resulted within 10 s in an insoluble, flexible hydrogel. A low molecular weight acidic molecule like trypan blue and various high molecular weight molecules such as bovine serum albumin (BSA), heparin and protamine were all retained within the hydrogel, while a low molecular weight basic molecule like toluidine blue was rapidly released from the hydrogel. In the present work, we examined the retaining capability of the chitosan hydrogel for growth factors and controlled release of growth factors from the chitosan hydrogel in vitro and in vivo. Fibroblast growth factor-1 (FGF-1), fibroblast growth factor-2 (FGF-2), vascular endothelial growth factor(165) (VEGF(165)), heparin-binding epidermal growth factor (HB-EGF) in phosphate buffered saline (PBS) were mixed with Az-CH-LA aqueous solution to form growth factor incorporated chitosan hydrogels. About 10-25% of the growth factor was released from a growth factor-incorporated chitosan hydrogel into PBS within the first day, after which no further substantial release took place. The growth factors interacted with Az-CH-LA molecules poly-ion complexation, and probably were unable to be released after the first day under the in vitro nondegradation conditions of the hydrogel. Although the FGF-1, FGF-2, and VEGF(165)-incorporated chitosan hydrogels on a culture plate significantly stimulated HUVEC growth, the stimulating activity of the growth factor-incorporated chitosan hydrogel was completely cancelled out by washing the hydrogel with PBS solution for 3 days or more. The stimulating activity on the HUVEC growth were however highly recovered by treating the washed growth factor-incorporated chitosan hydrogel during 7 days with chitinase and chitosanase to partly degrade the hydrogel, strongly suggesting that the growth factors within the hydrogel retained their biologically active forms. The chitosan hydrogel (100 microl) when implanted into the back of a mouse was biodegraded in about 10-14 days. When FGF-1- and FGF-2 incorporated chitosan hydrogels were subcutaneously implanted into the back of a mouse, significant neovascularization was induced near the implanted site of the FGF-1- and FGF-2-incorporated chitosan hydrogels. Furthermore, addition of heparin with either FGF-1 or FGF-2 into the hydrogel resulted in a significantly enhanced and prolonged vascularization effect. These results indicate that the controlled release of biologically active FGF-1 and FGF-2 with heparin is caused by biodegradation of the chitosan hydrogel, and subsequent induction of vascularization. PMID- 12579571 TI - Biosynthetic response of passaged chondrocytes in a type II collagen scaffold to mechanical compression. AB - To investigate the potential utility of mechanical loading in articular cartilage tissue engineering, porous type II collagen scaffolds seeded with adult canine passaged chondrocytes were subjected to static and dynamic compressions of varying magnitudes (0-50% static strain) and durations (1-24 h), and at different times during culture (2-30 days postseeding). The effects of mechanical compression on the biosynthetic activity of the chondrocytes were evaluated by measuring the amount of (3)H-proline-labeled proteins and (35)S-sulfate-labeled proteoglycans that accumulated in the cell-scaffold construct and was released to the medium during the loading period. Similar to published results on loading of articular cartilage explants, static compression decreased protein and proteoglycan biosynthesis in a time- and dose-dependent manner (each p < 0.005), and selected dynamic compression protocols were able to increase rates of biosynthesis (p < 0.05). The main difference between the results seen for this tissue engineering system and cartilage explants was in the amount of newly synthesized matrix molecules that accumulated within the construct under dynamic loading, with less accumulating in the type II collagen scaffold. In summary, the general biosynthetic response of passaged chondrocytes in the porous type II collagen scaffolds is similar to that seen for chondrocytes in their native environment. Future work needs to be directed to modifications of the cell-seeded construct to allow for the capture of the newly synthesized matrix molecules by the scaffold. PMID- 12579572 TI - Numerical simulations on fatigue destruction of ultra-high molecular weight polyethylene using discrete element analyses. AB - Ultra-high molecular weight polyethylene (UHMWPE) is a heterogeneous material composed of a networked substructure of grain boundary and grain aggregation. A new numerical model based on the discrete element method (DEM) was proposed to examine microscopic defect formation and propagation in UHMWPE. Numerical simulations were carried out using this model under two types of loading condition: unidirectional repetitive compression (simple loading) and bidirectional repetitive compression (switched loading). Subsurface defects were initiated and propagated in the vicinity of grain boundaries under both loading conditions. The defect propagation behavior was especially sensitive to grain boundary allocation under switched loading. An increase in defects was more rapid under switched loading than under simple loading. These numerical results showed qualitatively good agreement with experimental ones. It is suggested that the newly developed numerical method based on the DEM is a promising method to investigate fatigue behavior of a heterogeneous material such as UHMWPE under complicated loading conditions. PMID- 12579574 TI - Issue dedicated to the memory of Peter A. Kollman. PMID- 12579576 TI - Assessment of the molecular dynamics structure of DNA in solution based on calculated and observed NMR NOESY volumes and dihedral angles from scalar coupling constants. AB - To assess the accuracy of the molecular dynamics (MD) models of nucleic acids, a detailed comparison between MD-calculated and NMR-observed indices of the dynamical structure of DNA in solution has been carried out. The specific focus of our comparison is the oligonucleotide duplex, d(CGCGAATTCGCG)(2), for which considerable structural data have been obtained from crystallography and NMR spectroscopy. An MD model for the structure of d(CGCGAATTCGCG)(2) in solution, based on the AMBER force field, has been extended with a 14 ns trajectory. New NMR data for this sequence have been obtained in order to allow a detailed and critical comparison between the calculated and observed parameters. Observable two-dimensional (2D) nuclear Overhauser effect spectroscopy (NOESY) volumes and scalar coupling constants were back-calculated from the MD trajectory and compared with the corresponding NMR data. The comparison of these results indicate that the MD model is in generally good agreement with the NMR data, and shows closer accord with experiment than back-calculations based on the crystal structure of d(CGCGAATTCGCG)(2) or the canonical A or B forms of the sequence. The NMR parameters are not particularly sensitive to the known deficiency in the AMBER MD model, which is a tendency toward undertwisting of the double helix when the parm.94 force field is used. The MD results are also compared with a new determination of the solution structure of d(CGCGAATTCGCG)(2) using NMR dipolar coupling data. PMID- 12579577 TI - Free energy calculations for theophylline binding to an RNA aptamer: Comparison of MM-PBSA and thermodynamic integration methods. AB - We have applied the molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) method (J. Srinivasan, T. E. Cheatham, P. Cieplak, P. A. Kollman, and D. A. Case, Journal of the American Chemical Society, 1998, Vol. 120, pp. 9401-9409) to study the interaction of an RNA aptamer with theophylline and its analogs. The MM-PBSA free energy analysis provides a reasonable absolute binding free energy for the RNA aptamer-theophylline complex formation. Energetic analysis reveals that the van der Waals interaction and the nonpolar contribution to solvation provide the basis for the favorable absolute free energy of complex. This trend is similar to other protein-ligand interactions studied previously. The MM-PBSA method also ranks the relative binding energies of five theophylline analogs approximately correctly, but not as well as the more conventional thermodynamic integration calculations, which were carried out to convert theophylline into its analogs. The comparison of MM-PBSA with TI suggests that the MM-PBSA method has some difficulties with the first-solvation-shell energetics. PMID- 12579573 TI - Porous polymer scaffolds surface-modified with arginine-glycine-aspartic acid enhance bone cell attachment and differentiation in vitro. AB - This study was designed to determine if the surface modification of porous poly(lactic acid) (PLA) scaffolds would enhance osteogenic precursor cell (OPC) attachment, growth, and differentiation. A covalently grafted amino group ( NH(2)), poly(L-lysine) (PLL), and the peptide arginine-glycine-aspartic acid (RGD) were selected for the evaluation. The hypothesis was that surface modification would have a positive impact on cell-substratum interactions. The experiment was performed by OPC cells being placed on PLA films and scaffolds modified with NH(2), PLL, or RGD in tissue culture media. OPC attachment to PLA films was assessed after 24 h of incubation. The growth and differentiation of the adherent OPCs on porous PLA scaffolds were assessed after 14 and 28 days for alkaline phosphatase (APase) activity and calcium levels, both of which increase as OPCs differentiate into mature bone cells. All assays were accomplished in triplicate, and data were tested with post hoc orthogonal contrasts (i.e., Fisher's least significant difference) at p < or = 0.05. The PLA film surface modified with RGD showed better OPC cell attachment than the other films. The cells on the PLA scaffolds surface-modified with RGD also exhibited an increase in APase activity and calcium levels in comparison with those on other scaffolds. This difference was apparent at both time intervals and was especially evident in the tissue culture media containing an osteogenic supplement. The results of this study indicate that modifying the surface of PLA polymer scaffolds with RGD enhances bone cell attachment and differentiation and may improve their ability to regenerate bone tissue more efficiently in wound models. PMID- 12579578 TI - Computing the transition state populations in simple protein models. AB - We describe the master equation method for computing the kinetics of protein folding. We illustrate the method using a simple Go model. Presently most models of two-state fast-folding protein folding kinetics invoke the classical idea of a transition state to explain why there is a single exponential decay in time. However, if proteins fold via funnel-shaped energy landscapes, as predicted by many theoretical studies, then it raises the question of what is the transition state. Is it a specific structure, or a small ensemble of structures, as is expected from classical transition state theory? Or is it more like the denatured states of proteins, a very broad ensemble? The answer that is usually obtained depends on the assumptions made about the transition state. The present method is a rigorous way to find transition states, without assumptions or approximations, even for very nonclassical shapes of energy landscapes. We illustrate the method here, showing how the transition states in two-state protein folding can be very broad ensembles. PMID- 12579579 TI - The relaxed complex method: Accommodating receptor flexibility for drug design with an improved scoring scheme. AB - An extension of the new computational methodology for drug design, the "relaxed complex" method (J.-H. Lin, A. L. Perryman, J. R. Schames, and J. A. McCammon, Journal of the American Chemical Society, 2002, vol. 24, pp. 5632-5633), which accommodates receptor flexibility, is described. This relaxed complex method recognizes that ligand may bind to conformations that occur only rarely in the dynamics of the receptor. We have shown that the ligand-enzyme binding modes are very sensitive to the enzyme conformations, and our approach is capable of finding the best ligand enzyme complexes. Rapid docking serves as an efficient initial filtering method to screen a myriad of docking modes to a limited set, and it is then followed by more accurate scoring with the MM/PBSA (Molecular Mechanics/Poisson Boltzmann Surface Area) approach to find the best ligand receptor complexes. The MM/PBSA scorings consistently indicate that the calculated binding modes that are most similar to those observed in the x-ray crystallographic complexes are the ones with the lowest free energies. PMID- 12579580 TI - Breaking non-native hydrophobic clusters is the rate-limiting step in the folding of an alanine-based peptide. AB - The formation mechanism of an alanine-based peptide has been studied by all-atom molecular dynamics simulations with a recently developed all-atom point-charge force field and the Generalize Born continuum solvent model at an effective salt concentration of 0.2M. Thirty-two simulations were conducted. Each simulation was performed for 100 ns. A surprisingly complex folding process was observed. The development of the helical content can be divided into three phases with time constants of 0.06-0.08, 1.4-2.3, and 12-13 ns, respectively. Helices initiate extreme rapidly in the first phase similar to that estimated from explicit solvent simulations. Hydrophobic collapse also takes place in this phase. A folding intermediate state develops in the second phase and is unfolded to allow the peptide to reach the transition state in the third phase. The folding intermediate states are characterized by the two-turn short helices and the transition states are helix-turn-helix motifs-both of which are stabilized by hydrophobic clusters. The equilibrium helical content, calculated by both the main-chain Phi-Psi torsion angles and the main-chain hydrogen bonds, is 64-66%, which is in remarkable agreement with experiments. After corrected for the solvent viscosity effect, an extrapolated folding time of 16-20 ns is obtained that is in qualitative agreement with experiments. Contrary to the prevailing opinion, neither initiation nor growth of the helix is the rate-limiting step. Instead, the rate-limiting step for this peptide is breaking the non-native hydrophobic clusters in order to reach the transition state. The implication to the folding mechanisms of proteins is also discussed. PMID- 12579581 TI - Gaussian docking functions. AB - A shape-based Gaussian docking function is constructed which uses Gaussian functions to represent the shapes of individual atoms. A set of 20 trypsin ligand protein complexes are drawn from the Protein Data Bank (PDB), the ligands are separated from the proteins, and then are docked back into the active sites using numerical optimization of this function. It is found that by employing this docking function, quasi-Newton optimization is capable of moving ligands great distances [on average 7 A root mean square distance (RMSD)] to locate the correctly docked structure. It is also found that a ligand drawn from one PDB file can be docked into a trypsin structure drawn from any of the trypsin PDB files. This implies that this scoring function is not limited to more accurate x ray structures, as is the case for many of the conventional docking methods, but could be extended to homology models. PMID- 12579582 TI - Atomistic protein folding simulations on the submillisecond time scale using worldwide distributed computing. AB - Atomistic simulations of protein folding have the potential to be a great complement to experimental studies, but have been severely limited by the time scales accessible with current computer hardware and algorithms. By employing a worldwide distributed computing network of tens of thousands of PCs and algorithms designed to efficiently utilize this new many-processor, highly heterogeneous, loosely coupled distributed computing paradigm, we have been able to simulate hundreds of microseconds of atomistic molecular dynamics. This has allowed us to directly simulate the folding mechanism and to accurately predict the folding rate of several fast-folding proteins and polymers, including a nonbiological helix, polypeptide alpha-helices, a beta-hairpin, and a three-helix bundle protein from the villin headpiece. Our results demonstrate that one can reach the time scales needed to simulate fast folding using distributed computing, and that potential sets used to describe interatomic interactions are sufficiently accurate to reach the folded state with experimentally validated rates, at least for small proteins. PMID- 12579583 TI - Modeling HIV-1 integrase complexes based on their hydrodynamic properties. AB - We present a model structure of a candidate tetramer for HIV-1 integrase. The model was built in three steps using data from fluorescence anisotropy, structures of the individual integrase domains, cross-linking data, and other biochemical data. First, the structure of the full-length integrase monomer was modeled using the individual domain structures and the hydrodynamic properties of the full-length protein that were recently measured by fluorescence depolarization. We calculated the rotational correlation times for different arrangements of three integrase domains, revealing that only structures with close proximity among the domains satisfied the experimental data. The orientations of the domains were constrained by iterative tests against the data on cross-linking and footprinting in integrase-DNA complexes. Second, the structure of an integrase dimer was obtained by joining the model monomers in accordance with the available dimeric crystal structures of the catalytic core. The hydrodynamic properties of the dimer were in agreement with the experimental values. Third, the active sites of the two model dimers were placed in agreement with the spacing between the sites of integration on target DNA as well as the integrase-DNA cross-linking data, resulting in twofold symmetry of a tetrameric complex. The model is consistent with the experimental data indicating that the F185K substitution, which is found in the model at a tetramerization interface, selectively disrupts correct complex formation in vitro and HIV replication in vivo. Our model of the integrase tetramer bound to DNA may help to design anti integrase inhibitors. PMID- 12579584 TI - The molecular basis of the temperature- and pH-induced conformational transitions in elastin-based peptides. AB - Elastin undergoes an inverse temperature transition and collapses at high temperatures in both simulation and experiment. We investigated a pH-dependent modification of this transition by simulating a glutamic acid (Glu)-substituted elastin at varying pHs and temperatures. The Glu-substituted peptide collapsed at higher temperature than the unsubstituted elastin when Glu was charged. The charge effects could be reversed by neutralization of the Glu carboxyl groups at low pH, and in that case the peptide collapsed at a lower temperature. The collapse was accompanied by the formation of beta-turns and short distorted beta sheets. Formation of contacts between hydrophobic side chains drives the collapse at high temperature, but interactions between water and polar groups (Glu and main chain) can attenuate this effect at high pH. The overall competition and balance of the polar and nonpolar groups determined the conformational states of the peptide. Water hydration contributed to the conformational transition, and the peptide and its hydration shell must be considered. Structurally, waters near polar residues mainly formed hydrogen bonds with the protein atoms, while waters around the hydrophobic side chains tended to be parallel to the peptide groups to maximize water-water interactions. PMID- 12579585 TI - Simulation of the different biological activities of diethylstilbestrol (DES) on estrogen receptor alpha and estrogen-related receptor gamma. AB - We describe the application of the molecular dynamics (MD) and molecular mechanics-generalized Born/surface area (MM-GB/SA) approaches to the simulation of the different biological activity of diethylstilbestrol (DES) on two highly homologous nuclear receptors-estrogen receptor alpha (ER-alpha) and estrogen related receptor gamma (ERR-gamma). DES exerts an agonistic effect against ER alpha and an antagonistic effect against ERR-gamma. Using the x-ray crystal structures of ER-alpha in the canonical agonist bound form (PDB code: 3ERD) and antagonist bound form (PDB code: 3ERT), ERR-gamma homology models have been constructed for the receptor in two different conformations. MM-GB/SA binding free energy calculations of DES in the ER-alpha and ERR-gamma structures suggest that DES exhibits a greater free energy of binding in the agonist bound conformation of ER-alpha, while the antagonist bound conformation is preferred for ERR-gamma. Further dissection of the free energy contributions coupled with calculation of the ligand binding pocket volume suggests that the van der Waals interactions for DES within the smaller binding pocket volume of ERR-gamma are less favorable and this is the main factor for DES antagonism in ERR-gamma. This approach has potential general applicability to the prediction of the biological activity of nuclear receptor ligands. PMID- 12579586 TI - 2002 Exercise and Physical Activity Conference, St Louis, Missouri: exercise and arthritis "we know a little bit about a lot of things em leader ". PMID- 12579587 TI - Use of complementary and alternative medicine by pediatric rheumatology patients. AB - OBJECTIVE: To assess how often the patients in The Hospital for Sick Children's rheumatology clinic use complementary and alternative medicine (CAM), and the type of CAM used. METHODS: Cross-sectional descriptive survey study in a tertiary care referral center. RESULTS: We found that 90 (64%) of 141 respondents used at least 1 form of CAM; 45 (50%) of these 90 used more than 1 form. Duration of illness was positively associated with the use of CAM; the difference in the mean length of illness between users (4.12 +/- 3.60 years) and nonusers (2.82 +/- 3.23 years) was 1.3 years (P = 0.04). Of the 41 respondents with more than 1 illness, 32 used CAM, compared with 57 of 96 who had only 1 illness (P = 0.036). CONCLUSION: Health care providers must be aware of their patients' possible use of CAM, especially those with more than 1 illness. PMID- 12579588 TI - Disease activity score for children with juvenile dermatomyositis: reliability and validity evidence. AB - OBJECTIVE: To assess the reliability and validity of the Disease Activity Score (DAS), an instrument used to evaluate children with juvenile dermatomyositis (JDM). METHODS: Psychometric study of internal consistency, reliability, rater agreement, and the relationship with measures of muscle strength and disability was conducted. RESULTS: The DAS ratings are internally consistent (reliability = 0.89) and describe a wide range of disease activity. The pediatric rheumatologists in this study agree on the presence of most of the disease indicators. Their disagreements tend to cancel each other, resulting in highly correlated (r = 0.79) overall measures across raters. Estimates of muscle weakness using the DAS and ratings of muscle strength obtained independently from therapists are highly related (r = -0.77), but estimates of disease activity and disability are weakly related (r = 0.20). CONCLUSION: The DAS exhibits evidence of good reliability and validity. The combination of skin and muscle strength assessments makes this easily administered instrument a useful addition in the evaluation of children with JDM. PMID- 12579589 TI - Evidence for aerobic insufficiency in women with systemic Lupus erythematosus. AB - OBJECTIVE: To determine if fatigue is associated with diminished aerobic capacity in women with systemic lupus erythematosus (SLE). METHODS: Eighteen women (age 35 +/- 9 years) with mild SLE (Systemic Lupus Activity Measure = 3.1 +/- 2.1) and 16 healthy but sedentary controls (age 38 +/- 8 years) completed peak treadmill exercise tests to determine aerobic capacity and Fatigue Severity Scales to quantify the severity of fatigue. Measures of oxygen consumption (VO(2)) were recorded during the treadmill tests. RESULTS: Peak VO(2) was lower in patients with SLE (19.2 +/- 4.4 ml/kg/minute) as compared with controls (27.4 +/- 4.7 ml/kg/minute) and expected values (30.7 +/- 3.1 ml/kg/minute; P < 0.0006 versus controls and P < 0.0001 versus expected). Functional aerobic impairment was observed in 14 of the 18 patients with SLE. In patients with SLE, ventilatory threshold, a marker for the onset of lactic acidemia, was observed at the lowest energy requirement for instrumental activities of daily living. Peak VO(2) in the patients with SLE was similar to the highest energy requirements for instrumental activities of daily living, leaving little or no reserve for more intense occupational and recreational activities. Peak VO(2) was significantly higher (P < 0.0001) than the activity of daily living requirements in controls, providing a substantial energy reserve. Fatigue severity score (FSS) was 5.0 +/- 1.4 in patients with SLE, with 14 of the 18 patients having scores above 4.0, a score indicating that fatigue severity limited physical activity. Of the 14, 12 had functional aerobic impairment. An FSS of greater than 4.0 was not observed in controls (mean = 2.5 +/- 0.7). CONCLUSION: In women with SLE, aerobic capacity was diminished to levels that were insufficient for engaging in activities of daily living and below those expected to result from physiologic deconditioning. This functional aerobic impairment was strongly correlated with the perception of severe, activity-limiting fatigue. PMID- 12579590 TI - Quality of well-being in older people with osteoarthritis. AB - OBJECTIVE: To examine the sensitivity of the Quality of Well-Being Scale (QWB) as a measure of health-related quality of life (HRQOL) in people with osteoarthritis (OA). METHODS: The QWB was administered, along with the Arthritis Impact Measurement Scale (AIMS) and other health measures. Health care utilization data were also obtained. RESULTS: People with OA had a mean QWB score of 0.643. The QWB scores were significantly correlated with total AIMS scores, self-rated health status, health care costs, depression scores, and most AIMS subscales. In addition, changes in QWB scores after 1 year were significantly correlated with changes in total AIMS scores and some AIMS subscales. CONCLUSION: The QWB appears to be a useful and sensitive generic, utility-based measure of HRQOL in people with OA. PMID- 12579591 TI - Isometric muscle force measurement for clinicians treating patients with osteoarthritis of the knee. AB - OBJECTIVE: To evaluate the discriminant validity and, for clinicians, the test retest reliability of isometric force in patients with knee osteoarthritis (OA). METHODS: Mean isometric muscle strength data collected for 113 patients with knee OA were compared with published normative data for 131 asymptomatic subjects. RESULTS: Patients with knee OA, ages 60-79 years, could attain only 40-53% of the knee extensor force and 35-46% of the knee flexor force generated by their age- and sex-matched peers. One-week test-retest intraclass correlations ranged from 0.79 to 0.95. Random measurement error demonstrated a favorable "signal-to-noise" ratio. CONCLUSION: The similar decrease in knee extensor and knee flexor strength demonstrated by patients with knee OA compared with their age- and sex-matched asymptomatic peers suggests that strategies should be directed at both muscle groups to optimally limit joint damage. Isometric muscle force measurement can be a useful assessment tool for clinicians. PMID- 12579592 TI - Preoperative predictors of locomotor ability two months after total knee arthroplasty for severe osteoarthritis. AB - OBJECTIVE: To identify preoperative predictors of locomotor ability 2 months after total knee arthroplasty (TKA). METHODS: Sixty-five participants scheduled for a first TKA were recruited. The dependent variable was the distance covered during the 6-minute gait test 2 months after TKA. The independent variables, measured before surgery, were grouped into 3 categories: 1). personal; 2). organic system, and 3). capability variables. Multiple regression analyses were conducted to determine the best predictors of the dependent variable. Modified cross-validation of the model combining predictors from the three categories was obtained with 10 random samples derived from the original cohort by resampling with replacement. RESULTS: Predictors were identified in the 3 categories: 1). gender, number of comorbidities, body mass index, 2). knee pain, flexion and strength, lower limb mechanical power, and 3). preoperative 6-minute gait distance. When these variables were modeled together only the preoperative 6 minute gait distance and knee pain and flexion remained significant (adjusted R(2) = 0.66). CONCLUSION: Using variables easily measured before surgery, it is possible to predict with good accuracy locomotor ability 2 months after TKA. PMID- 12579593 TI - Use of the Trust in Physician Scale in patients with rheumatic disease: psychometric properties and correlates of trust in the rheumatologist. AB - OBJECTIVES: To assess the psychometric properties of the Trust in Physician Scale and to identify variables associated with patients' trust in their rheumatologist. METHODS: Analyses of self reported data from 713 patients with rheumatoid arthritis, osteoarthritis, or fibromyalgia. Study variables included the Trust in Physician Scale, a decision-making question, a medical skepticism measure, and demographic and health-related measures. Internal consistency and construct validity were assessed using correlational analyses and factor analysis. A regression analysis was conducted to identify factors associated with trust in the rheumatologist. RESULTS: Internal consistency of the scale was high (Cronbach's alpha = 0.87). Scale items also loaded on a single factor. Construct validity was supported by inverse correlations between higher trust scores and both skepticism and independent decision making. Decreased trust was associated with older age, minority status, higher education, diagnosis of fibromyalgia or osteoarthritis, and poorer health. CONCLUSION: The Trust in Physician Scale is appropriate for patients with rheumatic disease. Several patient characteristics appear to be associated with lower trust in the rheumatologist. PMID- 12579594 TI - The assessment of rheumatoid arthritis and the acceptability of self-report questionnaires in clinical practice. AB - OBJECTIVE: To assess the acceptability of self-report questionnaires (SRQ) in clinical practice and to understand the value that rheumatologists give to various assessment methods in rheumatoid arthritis. METHODS: Rheumatologists who completed a training course in the use of SRQ in clinical trials and clinical practice used the SRQ in their practices. Six months later 221 rheumatologists completed a survey regarding their experiences in assessing rheumatoid arthritis and in the use of SRQ. RESULTS: Prior to the start of the program, 18% of rheumatologists used self-report questionnaires, 6 months later, 48% were using SRQ in their practices. Rheumatologists who did not use questionnaires placed less value on all assessment methods, and particularly on questionnaire assessments of function and pain. They also were more likely to report that questionnaires were difficult to use, not accepted by staff, were too long, and that they had limited staff. Rheumatologists who used the questionnaires reported none of these difficulties and were satisfied with the benefits provide by SRQ. When assessment measures were ranked, rheumatologist ranked ACR 20, radiography, and erythrocyte sedimentation rate/c-reactive protein as the least important in both clinical trials and in clinical practice, and they rated swollen and tender joint counts followed by SRQ as the most useful assessment tools. CONCLUSION: SRQ are well received by rheumatologists, and following a training program almost 50% continued to use SRQ in their practices. Those who used questionnaires were generally more positive about assessments and had little difficulty in the technical aspects of administration, scoring, and interpretation. PMID- 12579595 TI - Costs and predictors of costs in rheumatoid arthritis: a prevalence-based study. AB - OBJECTIVE: To analyze the annual cost of rheumatoid arthritis (RA) and its predictive factors. METHODS: Data were obtained from a 12-month retrospective cohort of 201 RA patients, randomly selected from a rheumatology registry, through a structured interview and records of the Central Information System of the hospital. Results were divided into direct, indirect, and total costs in 2001 US dollars. A sensitivity analysis was performed. Multiple linear regression models for the different types of costs were carried out. RESULTS: The total cost was US dollars 2.2 million per year, with a cost attributable to RA of US dollars 2.07 million per year. The average cost per patient was US dollars 10419 per year (ranging from US dollars 7914 per patient per year in the best scenario to US dollars 12922 per patient per year in the worst case). Direct costs represent nearly 70% of total costs. We found an average increment in total costs of US dollars 11184 per year per unit of Health Assessment Questionnaire (HAQ) score (P < 0.0001) and an average annual increment of US dollars 621 per year of disease (P < 0.0001). After adjustment, the HAQ score, inability to perform housework tasks, and being permanently disabled for work were the only predictors of high costs. CONCLUSION: Our data show a remarkable economic impact of RA over society and link the costs of the disease to its consequences in terms of functional disability, work disability, and housework disability. PMID- 12579596 TI - Effects of dynamic strength training on physical function, Valpar 9 work sample test, and working capacity in patients with recent-onset rheumatoid arthritis. AB - OBJECTIVE: To study the impact of 24 months of strength training on the physical function of patients with early rheumatoid arthritis (RA). METHODS: Seventy patients were assigned to either the strength training (experimental) group (n = 35) or the control group (n = 35). Patients in the experimental group performed strength training for 24 months, and control patients were instructed to perform range of motion exercises. Maximal strength of the knee extensors, trunk flexors, and extensors, as well as grip strength were recorded with dynamometers. Disease activity was assessed by the erythrocyte sedimentation rate and Ritchie's articular index, joint damage was determined by the Larsen x-ray index, and functional capacity was assessed using the Valpar 9 test and the Stanford Health Assessment Questionnaire (HAQ). The employment status of each patient was recorded. RESULTS: In the experimental group, strength training led to significant increases (19-59%) in maximal strength of the trained muscles. Such increases in the control group varied from 1% to 31%. There was a clear training effect on muscular strength in favor of the experimental group, but significant improvements in the HAQ indices as well as in the Valpar 9 test were seen also in control patients. Results of the Valpar 9 and the HAQ were statistically significantly better in patients who remained gainfully employed compared with patients who retired preterm during followup. However, compared with patients who remained in the work force, patients who retired were older, and their work was physically more demanding. CONCLUSION: As expected, strength training led to increased muscle strength, but this increase did not correlate with improved physical function as assessed by the Valpar 9 work sample test. The increased muscle performance did not prevent a substantial proportion of patients from retiring preterm. The 2 items from the Valpar 9 test that were applied were not sensitive enough to differentiate the patients according to their working status. PMID- 12579597 TI - Development of a new instrument for rheumatoid arthritis: the Cedars-Sinai Health Related Quality of Life instrument (CSHQ-RA). AB - OBJECTIVE: To update and complement existing instruments, we developed a multidimensional disease-specific instrument, intended to reflect the impact of rheumatoid arthritis (RA) with modern treatment options on patient's Health Related Quality of Life (HRQOL). METHODS: Items were developed from a systematic review of published HRQOL measures and transcripts of RA patient focus groups. Items were refined by an expert panel and administered to 350 patients for psychometric testing. RESULTS: The systematic review identified 228 potential items, and the focus group transcripts identified 96 additional items. Expert review and pilot testing resulted in an initial 58-item instrument. Twenty-six items were excluded due to floor/ceiling effects, poor response rates, or high item-item correlations. Factor analysis identified a 5-factor structure (eigenvalues >or=1). Multi-trait scaling performed on both completed surveys confirmed the 5 sub-scale structure (Cronbach's > 0.87). CONCLUSION: The CSHQ-RA consists of 33 items that address 5 HRQOL domains, each with high internal consistency. Additional testing will assess the instrument's validity and responsiveness. PMID- 12579598 TI - Autoantibodies against B23, a nucleolar phosphoprotein, occur in scleroderma and are associated with pulmonary hypertension. AB - OBJECTIVE: To determine whether the abundant nucleolar phosphoprotein B23 is a target of autoantibodies in scleroderma, and to examine the clinical phenotype associated with these antibodies. METHODS: Ninety-two randomly selected scleroderma sera were screened by enzyme-linked immunosorbent assay against recombinant human B23. Demographic, clinical, and serologic parameters associated with B23 autoantibody status were examined. RESULTS: We demonstrated that autoantibodies against B23 occur in approximately 11% of sera obtained from patients with scleroderma. B23 seropositivity was related to pulmonary hypertension, antifibrillarin antibody, anti-RNP antibody, and decreased lung capacity. In multivariate analysis, B23 autoantibodies remained strongly associated with moderate-to-severe pulmonary hypertension and antifibrillarin antibodies. CONCLUSION: These data unite B23 with the group of nucleolar autoantigens targeted in scleroderma and thus focus attention on changes in the nucleolus that render its components immunogenic in this disease. The demonstration that antibodies to B23 are associated with an increased prevalence of pulmonary hypertension points to anti-B23 antibodies as a possible marker of a specific phenotype in scleroderma. PMID- 12579599 TI - Treatment of polyarteritis nodosa and microscopic polyangiitis with poor prognosis factors: a prospective trial comparing glucocorticoids and six or twelve cyclophosphamide pulses in sixty-five patients. AB - OBJECTIVE: Because the optimal cyclophosphamide (CY) treatment duration for severe polyarteritis nodosa (PAN) without virus infection and microscopic polyangiitis (MPA) has not been established, we conducted a trial to compare the effectiveness of 6 versus 12 CY pulses given in combination with corticosteroids (CS). METHODS: Sixty-five (18 PAN, 47 MPA) previously untreated patients were randomized to receive 12 (n = 34) or 6 (n = 31) CY pulses combined with CS. PAN and MPA were histologically proven or met ACR criteria. All patients presented >or=1 factor of severity according to the five factor score (FFS). CY pulses were administered every 2 weeks for 1 month, then every 4 weeks. The end point of the study was the number of events (relapses and/or deaths) occurring in each group, analyzed according to an intention-to-treat strategy. The outcome was evaluated by Cox proportional hazards analysis. RESULTS: The baseline characteristics were similar for both groups. The mean (+/- SD) followup was 32 +/- 21 months. Survival analysis showed a significantly lower relapse probability (P = 0.02; hazards ratio [HR] = 0.34) and higher event-free survival (P = 0.02, HR = 0.44) for the 12 CY-pulse group while the mortality rates were not significantly different (P = 0.47). CONCLUSION: These results suggest that 6 CY pulses are less effective than 12 CY pulses to treat severe PAN and MPA, particularly with respect to the risk of relapses. PMID- 12579600 TI - The costs of arthritis. PMID- 12579601 TI - Disability models: implications for arthritis exercise and physical activity interventions. PMID- 12579603 TI - Major public health benefits of physical activity. PMID- 12579604 TI - Physical activity levels among the general US adult population and in adults with and without arthritis. PMID- 12579605 TI - Correlates of physical activity: who's active and who's not? PMID- 12579607 TI - Moses: did he inherit the spondylarthritis of the Pharaohs? PMID- 12579609 TI - Comment on the "fibromyalgia" label: more cons than pros. PMID- 12579610 TI - Risk factors for positive minor salivary gland biopsy findings in Sjogren's syndrome and dry mouth patients: something new? PMID- 12579611 TI - Enthesitis of spondylarthritis can masquerade as Osgood-Schlatter disease by radiographic findings. PMID- 12579614 TI - Introduction to this issue: disability, public policy, and technology. PMID- 12579615 TI - The promise of the internet for disability: a study of on-line services and web site accessibility at Centers for Independent Living. AB - The Internet provides individuals with disabilities numerous tools to live independently. In the convenience of the home, a person can access an abundance of information, an electronic community, updates on the latest disability advocacy news, education through distance-learning classes, and on-line shopping for books, clothes, assistive technology, and a host of other consumer goods. Centers for Independent Living (CILs) are consumer-run, non-profit grassroots disability service organizations at the forefront of the disability rights movement. Providing services to individuals across the range of disabilities, CILs have begun to use the Internet as a complement to their traditional service delivery methods. This article examines the emerging trend of independent living services on the web. The investigation examines 200 CIL Internet sites across the United States during the period of April to August 2001. Information is collected and analyzed about how CILs are using the Internet to provide their services and programs. In addition, the article examines the technological accessibility of their web sites. Implications of the findings for CILs, consumers with disabilities, and disability policy are examined. PMID- 12579616 TI - Electronic doors to education: study of high school website accessibility in Iowa. AB - The Americans with Disabilities Act (ADA), and Sections 504 and 508 of the Rehabilitation Act, prohibit discrimination against people with disabilities in all aspects of daily life, including education, work, and access to places of public accommodations. Increasingly, these antidiscrimination laws are used by persons with disabilities to ensure equal access to e-commerce, and to private and public Internet websites. To help assess the impact of the anti discrimination mandate for educational communities, this study examined 157 website home pages of Iowa public high schools (52% of high schools in Iowa) in terms of their electronic accessibility for persons with disabilities. We predicted that accessibility problems would limit students and others in obtaining information from the web pages as well as limiting ability to navigate to other web pages. Findings show that although many web pages examined included information in accessible formats, none of the home pages met World Wide Web Consortium (W3C) standards for accessibility. The most frequent accessibility problem was lack of alternative text (ALT tags) for graphics. Technical sophistication built into pages was found to reduce accessibility. Implications are discussed for schools and educational institutions, and for laws, policies, and procedures on website accessibility. PMID- 12579617 TI - Technology for independence: a community-based resource center. AB - Despite the prominence of the disability civil rights model--with its values of inclusion and empowerment--the majority of social and policy research conducted to date has not sufficiently included the perspective of persons with disabilities in the research process and as uniquely qualified researchers themselves. This article describes a new project, "Technology for Independence: A Community-Based Resource Center" (CBRC). Over a five-year period, the CBRC will attempt to enhance community and consumer-directed disability organizations to design, implement, and disseminate research that promotes access to and use of assistive technology (AT). The CBRC will use strategies such as leadership training, participatory action research, technical assistance, web-assisted training, and annual symposia. A primary goal of the CBRC is to increase the capacity of community organizations to conduct research on AT that is scientifically rigorous and relevant to disability services, policy, and law. PMID- 12579618 TI - The verdict on jury trials for juveniles: the effects of defendant's age on trial outcomes. AB - With the progression to more adult-like policies and procedures for youth in the justice system, the right to a jury trial has been extended to young offenders. These youth would not be tried by a jury of their peers, however, but by a jury of adults. The concern is that adult jurors may hold negative attitudes about youth that might influence their decision making in a case involving a young defendant. Two studies examined whether and under what conditions defendant's age affects jurors' decisions about the guilt and sentencing of an accused. In study 1, data were gathered from two samples of jury eligible adults: one university sample and one public sample. Mock jurors read written transcripts of a trial involving a defendant who was presented as either 13, 17, or 25 years of age. Results indicated that the defendant's age had no effect on mock jurors' verdict or their ratings of defendant guilt. However, younger defendants were granted shorter sentences than the adult defendants. In study 2, mock jurors read the same trial presented in study 1 but were asked to deliberate about the case and render group verdicts. These group verdicts did not differ significantly by defendant's age. Age-related themes that emerged from group deliberations were identified, and results indicated that age tended to be used as a mitigating factor in favor of youth rather than against them. These findings are discussed in terms of their implications for youth justice policy and practice. PMID- 12579619 TI - Erotomanic stalking in evolutionary perspective. AB - Erotomania, the delusion of being loved by another person, comprises marked sex differences concerning prevalence rates and behavior. Whereas traditional psychiatry has considered erotomania to be almost entirely restricted to women, recent studies have revealed that criminal offenses associated with the condition occur much more frequently in men. The main hypothesis of this article is that these findings may be explained in terms of evolutionary theory. Erotomania, accordingly, may be viewed as a pathological variant of a specific sexual strategy that evolved under selection pressures of the human environment of evolutionary adaptedness. The overt behavior is related to the pursuit of long term mating, its potentially beneficial effect on inclusive fitness of the individual, and disparate strategies of the sexes to ensure sexual fidelity of the potential partner. Therefore, the evolutionary approach provides a plausible explanation as to why forensically relevant erotomania prevails in men. The pathological process of delusional misinterpretation of perceived signals from the social environment itself may result from poor reality testing due to a failure of social meta-cognition. The evolutionary perspective may provide additional insights into the nature of sex-specific behaviors and may improve our understanding of forensically relevant behaviors. PMID- 12579620 TI - Predicting criminal justice outcomes with the Psychopathy Checklist and Lifestyle Criminality Screening Form: a meta-analytic comparison. AB - Studies that have used either the Psychopathy Checklist--Revised (PCL-R) or the Lifestyle Criminality Screening Form (LCSF) to predict criminal justice outcomes (disciplinary adjustment and recidivism) were subjected to meta-analysis and compared. Analogous results were obtained with the PCL-R and LCSF even though the PCL-R takes 2-3 hours and the LCSF 10 minutes to complete. It is concluded that pending further evaluation in which these assessment measures are directly compared, the cost-effectiveness of the LCSF makes it an attractive alternative to the PCL-R in situations where risk of future offender disciplinary maladjustment or recidivism is of principal concern. PMID- 12579621 TI - Assessment of competency for execution: professional guidelines and an evaluation checklist. AB - The issue of whether mental health professionals should be involved in conducting evaluations of competency for execution is a topic that has elicited controversy and heated debate. This article picks up at a point beyond the controversy and addresses issues of professionalism and the objective assessment of competency for execution. Specifically, this article identifies professional standards for conducting competence for execution (CFE) evaluations, describes current practices in this area, and provides an interview checklist that can be used as an evaluation guide by involved professionals. PMID- 12579622 TI - The emerging debate over the shape of informed consent: can the doctrine bear the weight? AB - Informed consent has traditionally focused on treatment-related issues. However, since the mid-1990s, courts have debated whether informed consent should be stretched to accommodate other concerns. For example, some courts have considered whether economic limitations on treatment availability must be made known to a patient as part of the informed consent process. Other courts have considered whether characteristics of the treatment provider (e.g., experience with a particular procedure) should be disclosed as part of informed consent. Consideration of these issues turns in large part on whether the information in question would be considered "material" to a decision to accept or reject treatment. This article discusses these developments, and suggests that expanding informed consent beyond treatment-related issues in some circumstances may erode trust in the clinical relationship, thus undermining one of the central values of the informed consent doctrine. It concludes with some suggestions on how research could inform this debate. PMID- 12579623 TI - Urodynamics. PMID- 12579624 TI - Frequency-volume chart: the minimum number of days required to obtain reliable results. AB - AIMS: There is wide variation in the number of days necessary to maintain a diary and still furnish reliable data on which to base a sound clinical assessment. Estimates range from 1 day to 2 weeks, 7 days probably being the criterion standard. The goal of this retrospective study was to evaluate how much the 7-day period could be shortened without compromising the reliability of data. METHODS: Various lengths of frequency-volume (FV) charts (from 1 day to 6 days) were compared with the standard 7-day charts on 14 FV parameters. RESULTS: Overall results show that a 4-day dairy is nearly identical to the 7-day chart (most r > or = 0.95). Results of the 1-, 2-, and 3-day charts were frequently different statistically from the 7-day chart, whereas comparison of the 4-day chart with the 7-day chart showed no statistically significant differences. In addition, results of 4-day FV charts from a new control cohort showed no significant differences from the 7-day charts of the main cohort. CONCLUSIONS: In conclusion, our study indicates that the 4-day chart is as reliable as the 7-day chart. This reduction in the length of time, although easier for the patients, does not compromise the diagnostic value of the FV charts. PMID- 12579625 TI - Measuring health-related quality of life in women with urogenital dysfunction: the urogenital distress inventory and incontinence impact questionnaire revisited. AB - AIMS: Symptoms of urogenital dysfunction are known to negatively affect health related quality of life in women. To assess effectiveness of treatment, it is currently recommended to include measurements of quality of life in outcome analysis. One of the questionnaires that is commonly used is the combination of the Urogenital Distress Inventory (UDI) and Incontinence Impact Questionnaire (IIQ). Unfortunately, the validity of the UDI and IIQ has only been tested in highly selected subgroups of female patients. Therefore, it is unclear whether this questionnaire is suitable for use in populations with different characteristics. METHODS: We analyzed the scale construction and validity of the UDI and IIQ in a random sample of 2,042 women, aged 20-70 years old and a clinical sample of 196 women. RESULTS: Our results show that the UDI can be divided into five subscales, namely discomfort/pain, urinary incontinence, overactive bladder, genital prolapse, and obstructive micturition. The internal consistency (Cronbach's alpha) ranged between 0.74 and 0.82. In addition to the original four subscales of the IIQ (mobility, physical, social, and emotional functioning), we identified a fifth subscale with four items about embarrassment. Internal consistency of these subscales ranged between 0.83 and 0.93. In addition to the internal consistency, we tested the criterion and construct validity of these new subscale division. CONCLUSIONS: We found these subscales to be reliable and of clinical use. It is recommended to use the revised UDI and IIQ in outcome analysis of treatments for urogenital symptoms in women. PMID- 12579626 TI - Overactive bladder symptoms: do we need urodynamics? AB - AIMS: The aim of our study was to determine whether the urodynamic diagnosis is useful in the management of women with symptoms of an overactive bladder (OAB). METHODS: Women with lower urinary tract symptoms, attending a tertiary referral urogynaecology clinic were studied. All women were fully evaluated, with history, urinary symptoms questionnaire, frequency-volume chart, vaginal examination, and videocystourethrography. Women with symptoms consistent with an overactive bladder (urinary frequency, urgency, and/or urge incontinence) were selected. Women with neurological disorders were excluded. Finally, urinary symptoms and urodynamic diagnosis were correlated. All terms and definitions are in accordance with the International Continence Society [Abrams et al., 1988, Scand J Urol Nephrol 114(Suppl):5-19.]. RESULTS: A total of 4,500 women 22-73 years of age were studied. Only 843 women (18.7%) could be classified as having an OAB. Of these, 457 women (54.2%) had urodynamically proven detrusor instability, whereas 386 women (45.8%) had a stable urodynamic trace. Sixty-eight (8.1%) of the women studied had postvoid residual greater than 100 mL. Of the 4,500 women studied, 1,641 (36.5%) had detrusor instability on laboratory urodynamics. Only 27.5% of these women (457 of 1,641) had OAB symptoms. CONCLUSIONS: Symptomatic diagnosis of OAB does not correlate with a urodynamic diagnosis of detrusor instability. The diagnosis of overactive bladder based on urinary symptoms underdiagnoses the condition of detrusor instability in a population of women suffering from lower urinary tract symptoms. Therefore, symptomatic diagnosis of OAB alone is not recommended. Our study suggested that urodynamic evaluation is mandatory in the management of the women with symptoms of an overactive bladder. PMID- 12579627 TI - Prevalence and correlation of urinary incontinence and overactive bladder in Taiwanese women. AB - AIMS: We randomly sampled a community-based, healthy population to evaluate the prevalence and correlation of urinary incontinence and overactive bladder. We also assessed the influences of these conditions on quality of life and associated risk factors. METHODS: Of the 1,581 women sampled (2.92% of registered female residents aged 20 years and older), 1,253 (79.1%) women were successfully interviewed by using the Bristol Female Urinary Tract Symptoms Questionnaire (r=0.87, P<0.05) and the Questionnaire of Impact index regarding the impact on quality of life (r=0.91, P<0.05). RESULTS: A total of 53.7% of the women sampled suffered from urinary incontinence and related symptoms. The prevalence of stress urinary incontinence, overactive bladder, and mixed incontinence, mutually exclusive of each, was 18.0%, 18.6%, and 17.1%, respectively, from the patients' perceptions. Judging by the criteria of the International Continence Society, the prevalence of the above three conditions was 4.3%, 2.4%, and 1.8%, respectively. Of the women who perceived storage symptoms, 21.1% experienced frequency, 12.6% had urgency, 25.5% had nocturia, and 9.1% had urge incontinence. The occurrence of stress urinary incontinence increased with age up to 65 years old (25% in 50 65-year-old cohort), and the number of overactive bladder conditions significantly increased in the elderly women (over 65 years old, 39.3%). Approximately two thirds of the incontinent women had restricted their social activities (due to worrying about wetting or leakage and no toilet facilities available), and approximately 19% of the incontinent women had an affected sexual life. However, only 27.1% of the women with urinary incontinence and related symptoms in this study had reported seeking medical services to solve these problems. CONCLUSIONS: The prevalence of urinary incontinence and overactive bladder in Taiwanese women is similar to that of Western women. PMID- 12579628 TI - Quest for standardisation of electrical sensory testing in the lower urinary tract: the influence of technique related factors on bladder electrical thresholds. AB - AIMS: The aim of this study is to determine the influence of technique-related factors on bladder electrical thresholds in order to establish more standardisation in electrical sensory testing in the lower urinary tract (LUT). METHODS: In 51 young healthy volunteers, electrical thresholds were determined in the bladder by using different techniques described in the literature. Within an individual, thresholds were repeatedly determined in the bladder while varying the position of the stimulating electrodes and the intravesical volume. RESULTS: The height of bladder electrical thresholds is influenced by the position of the stimulating electrodes and the volume of the intravesical fluid at the time of the examination. By filling the bladder, the sensitivity of the bladder toward electrical stimulation can be increased. CONCLUSIONS: Although the techniques to test electrical sensitivity have already proven to be a valuable tool in diagnosing the presence of neuropathy in the LUT, its full scope can only be evaluated after standardisation. Therefore, it is proposed that the position of the stimulating electrodes and the bladder volume at the time of the examination should be outlined in studies and standardised. PMID- 12579629 TI - Motor evoked potentials from the striated urethral sphincter: a comparison of concentric needle and surface electrodes. AB - AIMS: The aim of this study is to compare two different surface electrodes (intravaginal and intraurethral) with the concentric needle as a criterion standard and concomitant testing of the levator ani as a potential contaminator. METHODS: Thirty healthy women with a mean age of 52 years. Bland-Altman plots were used to compare recorded latencies from the various electrodes and muscles. RESULTS: There was no significant difference between the latencies recorded with the various electrodes from either muscle, but the limits of agreement were wide. However, the concentric needle electrodes proved more reliable with a higher rate of reproducible responses. CONCLUSIONS: We did not encounter major difficulties in using the concentric needle electrode for recordings in the external urethral sphincter, and this electrode was also more reliable than either surface electrode. Therefore, we advocate the use of concentric needle electrodes in future studies. PMID- 12579630 TI - Conditional stimulation of the dorsal penile/clitoral nerve may increase cystometric capacity in patients with spinal cord injury. AB - AIMS: To investigate the feasibility of conditional short duration electrical stimulation of the penile/clitoral nerve as treatment for detrusor hyperreflexia, the present study was initiated. METHODS: Ten patients with spinal cord injury, 4 women and 6 men, with lesions at different levels above the sacral micturition center had a standard cystometry performed. During a subsequent cystometry, conditional short duration electrical stimulation of the penile/clitoral nerve was performed as treatment for one or more detrusor hyperreflexic contractions. RESULTS: In all patients, at least one contraction (mean, 7.8; range, 1-16 contractions) was inhibited by the stimulations. The mean cystometric capacity was increased significantly by conditional electrical stimulation, from 210 mL in the control cystometries to 349 mL in the stimulation cystometries (P=0.016). The maximal detrusor pressure during the first contraction in the control cystometries was mean 51 cm H(2)O, whereas the maximal pressure of the first contraction in the stimulation cystometries was reduced to mean 33 cm H(2)O (P=0.045). CONCLUSIONS: The authors conclude that repeated conditional short duration electrical stimulation significantly increased cystometric capacity in patients with spinal cord injury. The increase was caused mainly by an inhibition of detrusor contractions. The need for a reliable technique for chronic bladder activity monitoring is emphasized, as it is a prerequisite for clinical application of this treatment modality. PMID- 12579631 TI - Does digital rectal examination affect uroflowmetry measurements? AB - AIMS: To determine whether digital rectal examination (DRE) would affect uroflowmetry parameters and whether these findings were not due to the circaidan changes. METHODS: A total of 79 male patients (59 in study and 20 in control group) were included in this study. First uroflowmetry was done at 10:00 am just before the genitourinary physical examination in the study group. Second and third uroflowmetry was performed at the 6th and 24th hour after the first uroflowmetry. The same uroflowmetry tests were done in a control group without DRE. Voided volume and maximum and average flow rate parameters of uroflowmetry were interpreted in our study. The relationship among first, second, and third uroflowmetry parameters were determined in the study and control groups. RESULTS: There were statistically no difference between the mean first, or 6th- or 24th hour uroflowmetry parameters of the study and control groups (P > 0.05). CONCLUSIONS: We could not find any effect of DRE on uroflowmetry parameters. Studies in a larger patient series would be appropriate for recommending DRE before or after uroflowmetry test. PMID- 12579632 TI - Incontinent ileovesicostomy in the management of neurogenic bladder dysfunction. AB - AIMS: To report outcome and urodynamic follow-up of incontinent ileovesicostomy in quadriplegic patients with neurogenic bladder. METHODS: Seven patients (five male, two female; mean age, 33.7 yr) with neurogenic bladder underwent ileovesicostomy for management of leakage or complications of chronic catheter drainage. Five had chronic indwelling catheters: three suprapubic and two urethral. Preoperatively, all had upper tract evaluation and videourodynamics. All seven patients had detrusor hyperreflexia. Preoperative detrusor leak point pressures averaged 42.7 cm H(2)O. Two females had intrinsic sphincteric deficiency from prolonged Foley catheter drainage. Ileovesicostomy involves isolation of a 15-20-cm segment of terminal ileum. The proximal 6-8 cm of this segment is opened on the antimesenteric border. The dome of the bladder is opened widely in a transverse manner and the proximal portion of the bowel is sutured onto the bladder. The distal portion of the ileum remains tubularized and becomes the stoma. RESULTS: There were no intraoperative complications. Operative time averaged 159 minutes. Associated procedures included removal of bladder calculus (n = 1), pubovaginal sling (n = 2), and Marshall Marchetti Krantz suspension (n = 1). Mean blood loss was <200 cc in six patients. Mean hospital stay was 8 days. Complications in two patients included: fascial stenosis requiring stoma revision (n = 1), wound infection (n = 1), and postoperative ileus (n = 1). Mean follow-up was 37.4 months. Postoperatively, mean detrusor leak point pressures were 16.7 cm H(2)O (P = 0.0061). Patient satisfaction is high with only one complaint of occasional difficulty fitting the appliance. CONCLUSIONS: Ileovesicostomy is an effective method of urinary drainage in quadriplegic patients. Detrusor leak point pressures were lowered, and upper tracts were preserved. No long-term complications were encountered. PMID- 12579633 TI - Effects of ATP-sensitive K+ channel openers and tolterodine on involuntary bladder contractions in a pig model of partial bladder outlet obstruction. AB - AIMS: To compare in vivo the efficacy, potency, and bladder-vascular selectivity of ATP-sensitive potassium channel openers (KCOs), YM934 and (-)-cromakalim to a muscarinic antagonist, tolterodine in a novel partial outlet obstructed pig model. METHODS: Partially obstructed female Landrace pigs were implanted with telemetry transmitters to allow the continuous measurement of intravesical, abdominal and arterial pressures. A subcutaneous port catheter was used to adjust bladder volume. Bladder and arterial pressure were simultaneously monitored under isoflurane anesthesia before and after increasing i.v. doses of test compounds. RESULTS: Under anesthesia, voiding was completely inhibited, but spontaneous, nonvoiding bladder contractions were observed with mean amplitude of 16 +/- 1 cm H(2)O, duration of 35 +/- 2 seconds, and intercontraction interval of 43 +/- 4 seconds (n = 25). YM934 and (-)-cromakalim both caused dose-dependent decreases in bladder contraction area under the curve (AUC) with effective doses to inhibit AUC by 35% of 3.6 and 14.9 nmol/kg, i.v., respectively. However, concomitant reductions in mean arterial pressure of 12 and 13% were also observed. Tolterodine did not inhibit spontaneous bladder contractions at doses up to 100 nmol/kg, i.v. corresponding to plasma concentrations up to 41 ng/mL. CONCLUSIONS: The superior efficacy of KCOs to inhibit spontaneous bladder contractions relative to tolterodine support the hypothesis that KCOs may provide an alternate therapeutic mechanism to treat symptoms of overactive bladder if bladder-vascular selectivity can be sufficiently improved. The minimally invasive model described herein appears useful in the preclinical evaluation of potential therapeutics targeted to treat the overactive bladder. PMID- 12579635 TI - Supraspinal N-methyl-D-aspartate receptor inhibition influences the micturition reflex and function of the upper urinary tract of anesthetized and conscious rats. AB - AIMS: To examine the effects of N-methyl-D-aspartate (NMDA) receptor antagonists on the frequency/volume (F/V) characteristics of micturition of conscious and anesthetized rats in relation to the mechanisms of renal urine output and pyelo ureteral transport function of the upper urinary tract. METHODS: Micturition F/V characteristics of 24 conscious female Sprague-Dawley rats, each weighting 160 180 g, were evaluated in a metabolic chamber. Control values of (F/V) were first obtained after administration of a 5-mL loading dose of saline solution and compared with intraperitoneal (i.p.) injection of 0.3-6.0 mg/kg of LY274614 (LY) and 0.3-40 mg/kg of dextromethorphan (DEX). Upper urinary tract studies were performed on two groups of urethane anesthetized rats (1.2 g/kg, s.c.). A group of 17 rats was used for intravenous (i.v.) injection of 1-30 mg/kg of LY and 1-10 mg/kg of DEX. In a second group of 12 rats, 0.2 mg/kg of LY and DEX were administered intracerebroventricularly (i.c.v.), by placing a catheter into the third ventricle. In anesthetized rats, continuous cystometrograms (CMG) were done while perfusing the renal pelvis with indigo carmine, and measuring pelvic and arterial pressure and as well as visualizing the transport of urine within the upper urinary tract by using videomicroscopic imaging. RESULTS: In conscious rats, the i.p. injection of LY and DEX produced a significant and dose-dependent increase in functional bladder capacity and a considerable increase in diuresis. In anesthetized rats, i.v. and i.c.v. administration of LY and DEX affected the micturition reflex, reflected as a significant increase in bladder capacity and preleakage pressure. Videomicroscopic imaging shows dilation of the ureter and a dilution of the dyed urine after the i.v. and i.c.v. injection of LY and DEX. The frequency of ureteral peristalsis and renal pelvic contractions were significantly decreased after i.v. and i.c.v. injection of LY as well as DEX. CONCLUSIONS: The present study shows that the NMDA receptor antagonists LY and DEX given systemically (i.v. and i.p.) or intracerebroventricularly influence the micturition reflex and modulate pyelo-ureteral motility as well as the rate of urine production. From the present study, it is suggested that NMDA receptors are directly involved in regulating pyelo-ureteral peristalsis as well as in the coordination of urine transport between upper and lower urinary tract. PMID- 12579634 TI - Focal hypoxia of the obstructed rabbit bladder wall correlates with intermediate decompensation. AB - AIMS: We showed that partial obstruction of the rabbit bladder outlet caused decreases in detrusor blood flow that were directly proportional to the level of decompensation present. Bladder decompensation is characterized by decreases in detrusor contractility, mitochondrial function, and sarco/endoplasmic reticulum calcium ATPase (SERCA) activity in obstructed rabbits. The current study was designed to create bladder decompensation and to relate its characteristic dysfunctions to the presence or absence of hypoxia in the obstructed rabbit bladder wall. Tissue hypoxia was visualized immunohistochemically after administration of a hypoxia probe in vivo. METHODS: Twelve New Zealand White rabbits were separated into two groups. The rabbits in group 1 received sham operations; the rabbits in group 2 received partial outlet obstructions by standard methods. Four weeks after surgery, each rabbit received an intraperitoneal injection of aqueous Hypoxyprobe-1, which forms protein adducts in cells having O(2) concentrations less than 14 microM. Two hours after injection, the rabbit was anesthetized and the bladder exposed through a midline incision. One full-thickness bladder strip was cut and immediately placed in fixative for immunohistochemical recognition and visualization of Hypoxyprobe-1 protein adducts. The remaining bladder was then excised, and three additional strips were cut for contractility studies. The remainder of the bladder was frozen for biochemical and slot-blot analyses. RESULTS: Bladder weight was increased fourfold after obstruction, and significant contractile and biochemical dysfunctions were observed that indicated an intermediate level of decompensation. Immunohistochemical visualization revealed focal areas of moderate to severe hypoxia in the detrusor smooth muscle (SM) and subserosal regions of these bladders. No hypoxia was observed in the obstructed bladder mucosa, consistent with the absence of biochemical dysfunction in this compartment, or in unobstructed bladders. Slot-blot analyses confirmed the presence of significant Hypoxyprobe-1-protein adducts in the detrusor of the obstructed bladder, whereas none were present in the control bladder detrusors. CONCLUSIONS: Partial outlet obstruction of rabbit bladders resulted in focal areas of moderate to severe hypoxia in the detrusor SM and subserosal regions concomitant with increased bladder mass, decreased contractile function, and selective metabolic dysfunctions of the SM consistent with an intermediate stage of decompensation. The metabolic characteristics of the normoxic mucosa were normal a were those of unobstructed bladders. PMID- 12579636 TI - Changes in renal hemodynamics and urodynamics in rats with chronic hyperoxaluria and after acute oxalate infusion: role of free radicals. AB - AIMS: The aim of this study was to evaluate possible changes in renal hemodynamic and urodynamic parameters in rats with chronic hyperoxaluria and after acute oxalate challenge. We also evaluated the possible association between free radical (FR) production, hyperoxaluria, and calcium oxalate (CaOx) calculi formation. METHODS: Chronic hyperoxaluria was induced by adding 0.75% ethylene glycol (EG) to the drinking water of male Wistar rats. After 7, 21, and 42 days of treatment, urinary biochemistry, oxalate levels, and lipid peroxides were measured. Kidney calculi were examined by polarizing microscopy. In the second part of the experiments, 1, 10, 20, and 30 mg kg(-1) hr(-1) oxalate was infused, by means of an intrarenal arterial catheter (IRA), into normal rats sequentially. Superoxide dismutase (SOD) infusion by means of IRA, in addition to oxalate, was also performed to check its influence on the altered renal function after oxalate infusion. In both the acute and chronic groups, renal blood flow (RBF), cortical microvascular blood flow (CMVBF), glomerular filtration rate (GFR), urine flow (UV), and urinary sodium excretion (U(Na)V) were measured, and chemiluminescence (CL) was examined in the renal venous blood. RESULTS: Levels of urinary lipid peroxides and enzymuria had increased since day 7, and increased the size of numbers of CaOx crystals in the kidney were noted beginning on day 21, but elevated CL was detectable only on day 7 after 0.75% EG treatment. Decreased UV and U(Na)V were noted in the 42-day EG group, although the 24-hr creatinine clearance values were normal in all experimental groups. On the other hand, RBF, GFR, and CMVBF were attenuated with elevated FR when the oxalate concentration was higher than 10 mg kg(-1) hr(-1) in the acute oxalate infusion group. With SOD pretreatment, the decreased RBF, GFR, and CMVBF could be reversed at 10 mg kg(-1) hr(-1) of oxalate, and be partially reversed at 20. FR also could be reduced significantly at 10 and 20 mg kg(-1) hr(-1) of oxalate. CONCLUSIONS: Decreased urine flow and sodium excretion were the main renal functions affected by chronic hyperoxaluria. However, that only the 42-day EG group had a decreased tubular function cannot be fully explained by the persistent tubular enzymuria and increased lipid peroxides that began on day 7 after EG treatment. With acute oxalate infusion, the major insult to renal function was renal hemodynamics. Pretreated SOD could reverse the attenuated hemodynamics and reduce the elevated FR partly, which suggested that FR is responsible for oxalate toxicity. PMID- 12579637 TI - Comment on "Physiologic bladder evacuation with selective sacral root stimulation: sinusoidal signal and organ-specific frequency," Neurourol urodyn 2002;21:80-91. PMID- 12579638 TI - The preliminary study of interferon-gamma gene transfection to human Tenon's capsule fibroblasts in vitro. AB - PURPOSE: To investigate the results of the interferon-gamma (IFN-gamma) gene transfer and transient expression in human Tenon's capsule fibroblast in vitro in order to find a way to gene therapy in vivo. METHOD: Using LipofectAMINE, IFN gamma gene was transferred in human Tenon's capsule fibroblasts with plasmid pcDNA3 IFN-gamma. Its mRNA transcription and protein expression were determined by RT-PCR and flow cytometry assay respectively. RESULT: The human Tenon's capsule fibroblasts transferred the IFN-gamma gene can express the IFN-gamma in transcription and protein level transiently. CONCLUSION: IFN-gamma gene can be transferred successfully and expressed expressed efficiently in human tenon's capsule fibroblast in vitro. PMID- 12579639 TI - Spatio-temporate expression of P53, Bcl-2 and Fas in the retina of RD, Rds and C3H mice. AB - PURPOSE: To investigate the expression status of p53, Fas and bcl-2 in the development of retinal degeneration in C3H, rd and rds mice. METHODS: Expression of p53, Fas and bcl-2 in the retina of rd, rds and C3H mice as well as in normal C3B mice in different periods was examined by immunohistochemical technique. RESULTS: The expression of Fas and p53 was not detected in normal C3B mice. Fas and p53 were expressed in the ganglionic layer in the early stage and then in the inner nuclear layer, while the scale and intense increased in the ganglionic layer. There is no difference of bcl-2 expression between the normal mice and mice with retinal degeneration. CONCLUSION: Fas and p53 may be involved in the retinal cell death in rd, rds, and C3B, but bcl-2 may not. It is unknown why p53 and Fas appeared first in ganglion layer but not in the outer nuclear layer where retinal cells death was noticed in early stage. PMID- 12579640 TI - Study on stereometric parameters of optic nerve head of normal, big-cupped disk and glaucomatous eyes using Heidelberg retina tomograph. AB - PURPOSE: To investigate the difference of stereometric parameters of optic nerve head between the normal subjects and patients with big-cupped disk and primary open angle glaucoma(POAG). METHODS: Twenty-two cases (44 eyes) of normal subjects, 17 cases (34 eyes) of patients with big-cupped disk and 19 cases (37 eyes) of patients with POAG underwent Heidelberg Retina Tomograph (HRT) examination to get topography images and stereometric parameters of optic nerve head. RESULTS: The stereometric parameters of optic nerve head of the normal, patients with big-cupped disk and POAG were 1) disk area (mm2): 1.995 +/- 0.501, 2.407 +/- 0.661 and 2.248 +/- 0.498; 2) cup area (mm2): 0.573 +/- 0.264, 1.095 +/ 0.673 and 1.340 +/- 0.516; 3) cup/disk ratio: 0.257 +/- 0.095, 0.428 +/- 0.176 and 0.589 +/- 0.195; 4) rim area (mm2): 1.461 +/- 0.328, 1.312 +/- 0.418 and 0.905 +/- 0.409; 5) cup volume (mm3): 0.108 +/- 0.073, 0.347 +/- 0.346 and 0.550 +/- 0.394; 6) rim volume (mm3): 0.421 +/- 0.111, 0.378 +/- 0.225 and 0.224 +/- 0.189; 7) mean cup depth (mm): 0.225 +/- 0.070, 0.327 +/- 0.105 and 0.424 +/- 0.148; 8) maxim cup depth (mm): 0.630 +/- 0.178, 0.732 +/- 0.170 and 0.871 +/- 0.245; 9) cup shape measure (mm): -0.1867 +/- 0.09, -0.108 +/- 0.068 and -0.02 +/ 0.164; 10) mean RNFL thickness (mm): 0.290 +/- 0.053, 0.236 +/- 0.131 and 0.174 +/- 0.134; 11) RNFL cross sectional area (mm2): 1.419 +/- 0.332, 1.263 +/- 0.606 and 0.920 +/- 0.687. Besides, there was no difference of disk area between big cupped disk group and POAG group, no difference of rim area, rim volume and RNFL thickness between normal group and big cupped disk group, statistics analysis showed that there was significant difference of other stereometric parameters between the normal group and big cupped disk group or big cupped disk group and POAG group. CONCLUSIONS: There is certain difference of optic nerve head between the normal subjects and big-cupped disk and patients with POAG. PMID- 12579641 TI - A comparative evaluation of Humphrey perimetry and the multi-channel pattern visual evoked potentials. AB - PURPOSES: To compare the multi-channel pattern visual evoked potentials to Humphrey perimetry in the assessment of central visual function in primary open angle glaucoma. METHODS: The multi-channel checkerboard reversal PVEPs waves to full-field and half-field stimulus of 25 normal persons and 74 patients with primary open angle glaucoma were recorded and analyzed, All patients were examined using Humphrey Field Analyzer. The area of visual field corresponding to the area of retina stimulated during multi-channel PVEPs testing were analysed, straight-line correlation and regression analyses of the various multi-channel PVEPs parameters and the total dB losses were performed. RESULTS: The multi channel PVEPs demonstrated a low detection rate compared with Humphrey perimetry in the early glaucoma, absolute latency and field loss were correlated in the late stage of glaucoma, and absolute amplitude and field loss were not correlated. CONCLUSIONS: In relation to signalling "early" loss the multi-channel PVEPs was inferior to Humphrey perimetry, in late loss of primary open angle glaucoma, multi-channel PVEPs can provide a valuable, objective complement to Humphrey perimetry. PMID- 12579642 TI - Measurement of episcleral venous pressure in normal Chinese people. AB - PURPOSE: To measure episcleral venous pressure (EVP) in normal Chinese people. METHODS: EVP was measured in 110 eyes of 74 subjects distributed in seven age groups from 10 to 80 years using episcleral venomanometer (EV-310). The differences in EVP values were analyzed with age and either eye. RESULTS: EVP values of the 110 eyes appeared as normal distribution. The value mean +/- SD was (9.11 +/- 1.22) mmHg, and the 95% normal value range was 6.73 mmHg -11.50 mmHg. No statistically significant difference of EVP was found in the seven age groups. And there was no statistically significant difference of EVP between the right and left eyes. CONCLUSIONS: The value mean +/- SD of EVP in Chinese normal eyes in 9.11 mmHg +/- 1.22 mmHg. The normal value of EVP does not vary with age and either eye. PMID- 12579643 TI - The changes of TGF-alpha, TGF-beta 1 and basic FGF messenger RNA expression in rabbit cornea after photorefractive keratectomy. AB - OBJECTIVE: To study the mechanism of haze formation and investigate the expression changes of transforming growth factor-alpha (TGF-alpha), transforming growth factor-beta 1(TGF-beta 1) and basic fibroblast growth factor (bFGF) mRNA in corneal epithelium and stroma after photorefractive keratectomy (PRK). METHODS: Sixteen white rabbits were randomly divided into 4 groups, and PRK was performed on each eye of 12 rabbits. The haze formation was examined under a slit lamp microscope at the 1st, 2nd and 3rd month after PRK, and the expressions of TGF-alpha, TGF-beta 1 and bFGF mRNA were detected with in situ hybridization. RESULTS: The corneal haze formed at the 1st month after PRK. The most prominent haze formation was observed at the 2nd month, and declined gradually at the 3rd month after ablation. TGF-alpha mRNA expression was presented on the normal corneal epithelium and not on the corneal stroma. TGF-beta 1 and bGFG mRNA were expressed by both corneal epithelium and stroma. The capacities for cornea tissue expression of three growth factors mRNA increased after PRK, and the peaks appeared on the 1st, 2nd month. The extent for expressions of three growth factors related proportionally to the haze formation. CONCLUSION: Three growth factors took part in promoting corneal wound healing after PRK, and might contribute to corneal haze formation and development. PMID- 12579645 TI - Lens epithelial cell proliferation and cell density in human age-related cataract. AB - PURPOSE: To discuss the potential effect of the lens epithelial cell proliferation in age-related cataract. METHODS: In vitro cell proliferation was assayed by MTT method to evaluate the lens epithelial cell density, index, and proliferation capacity in normal lens and all kinds of age-related cataract. Capsulotomy specimens from all kinds of patients who underwent cataract phacoemulsification extraction surgery were compared with the lens epithelial specimens from non-cataract lenses of Eye Bank eyes. RESULTS: Lens epithelial cell density of central anterior capsule (LECD) in female normal lens was higher than that in male, LECD in nuclear cataract (> NIII) was higher than that in normal lens, but in the mature cortical cataract, LECD was lower. Mitotic index of three kinds of age-related cataracts in vivo had no statistical difference, neither did cell proliferation capacity of cultivated cells in vitro. CONCLUSION: The individual difference of lens epithelial cell density and proliferation capacity in vivo may be an important underlying cause for senile cataract in the cellular level, especially for nuclear cataract. PMID- 12579644 TI - Clinical observation of 126 cases of sutureless phacoemulsification with PMMA intraocular lens implantation. AB - OBJECTIVE: To observe the safety and effect of sutureless phacoemulsification with PMMA intraocular lens (IOL) implantation. METHODS: One-hundred and twenty six cases (126 eyes) of sutureless phacoemulsification with PMMA intraocular lens (IOL) implantation were retrospectived and evaluated. The surgeries were performed through a limbal tunnel incision. PMMA IOLs with 5.5 mm or 6.0 mm in diameter were implanted. RESULTS: The follow-up was 3-20 months. Visual acuity of 0.5 or better was obtained in 120 cases (95.2%), and that of 1.0 or better was obtained in 89 cases (70.6%). Intra-operative complications included posterior capsule rupture in 9 cases (7.1%) and iris injury in 7 cases (5.6%). Postoperative complications included early corneal edema in 96 cases (76.2%), transient intraocular pressure elevation in 5 cases (4.3%), remnant of cortex in 1 case and leakage of incision in 1 case. All were properly managed with good results. CONCLUSION: Sutureless phacoemulsification with PMMA IOL implantation is safe even in grass-root hospitals where cataracts are harder. PMID- 12579646 TI - The effect of liposome encapsulated daunorubicin on rabbit eyes after extracapsular lens extraction. AB - PURPOSE: To investigate the effect of liposome encapsulated daunorubicin (DNR) on rabbit eyes when it was used in prevention of posterior capsule opacification (PCO). METHODS: The liposome encapsulated DNR was prepared by modified freeze thawing method. Each eye was injected with 0.1 ml liposomes (0.2 mg/ml and 20 micrograms/ml DNR) into the capsular bag during the extracapsular lens extraction (ECLE) in 10 rabbit eyes respectively. The phosphate buffer solution (PBS) was injected as control. Besides biomicroscope observation and histology examination of all eyes, the concentration of DNA in aqueous humor was also determined by high performance liquid chromatography (HPLC). RESULTS: The morphology of liposome encapsulated DNR were similar to the blank liposome with round or ellipse shape. The encapsulated effeciency of liposome encapsulated DNR was 45.1%. The inflammatory response was much more severe both in 0.2 mg/ml and 20 micrograms/ml DNR group than the control after liposome injection. All eyes in DNR group were showed severe corneal edema and opacity and lasted for 4 to 8 weeks. Histopathological study revealed that lens epithelial proliferation occured by 4 weeks and Soemmering's ring developed by 8 weeks in control; However, in DNR group eye only a few LECs remained in the bag and there were obvious corneal edema and eosinophiles infiltration. Corneal endothelial cells were severely lost. CONCLUSION: The results suggest that liposome encapsulated DNR could inhibit metaplasia and proliferation of remnant lens epithelial cells after ECLE in rabbit eyes, but the severe toxicity must be avoided by further study. PMID- 12579647 TI - Growth inhibition, induction of apoptosis by green tea constituent (-) epigallocatechin-3-gallate in cultured rabbit lens epithelial cells. AB - PURPOSE: To evaluate effect of green tea extract (-)-Epigallocatechin-3-gallate (EGCG) in cultured rabbit lens epithelial cells in order to pave a new way to postcapsular opacity (PCO) prevention. METHODS: Cell survival rate was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) coloimetric assay. Cell apoptosis was detected by electron microscopy, Hochest 33258 stain and flow cytometer. DNA fragment was detected using agarose gel electrophoresis. RESULT: Proliferation of the cultured rabbit lens epithelia cells was inhibited by EGCG in a dose and time dependent manner. Morphologic study showed that the cells became shrunk, round shaped with their nuclei condensed and broken. Apoptotic bodies were also seen under electron microscope and in Hochest 33258 stain assay 24 hours after EGCG was added to the medium. DNA ladders were shown in agarose gel electrophoresis. In flow cytometry assay, apoptosis peak was also evident. CONCLUSION: Green Tea Constituent(-) Epigallocatechin-3-gallate could inhibit cultured rabbit lens epithelial cells proliferation by inducing their apoptosis in the concentration used by us, which indicates that it is possible to prevent PCO by using herb extract. PMID- 12579648 TI - Detection of glutamate and gamma-aminobutyric acid in vitreous of patients with proliferative diabetic retinopathy. AB - PURPOSE: To study the levels of glutamate and gamma-aminobutyric acid (GABA) in vitreous of patients with proliferative diabetic retinopathy (PDR) and to investigate their roles in retinal ischemia. METHOD: Vitreous samples were collected from 25 patients (27 eyes) with PDR and 14 patients (14 eyes) with idiopathic macular hole. Glutamate and GABA detection were performed by high performance liquid chromatography (HPLC). RESULTS: Patients with PDR had significantly higher concentrations of glutamate and GABA than the control group. The glutamate level has a significantly positive correlation with GABA level. CONCLUSION: Detection of glutamate and GABA in vitreous provides biochemical support for the mechanism and treatment of ischemic retinal damage in patients with PDR. PMID- 12579649 TI - Vitro culture and immunohistochemical identification of astrocytes of infantile optic nerve. AB - OBJECTIVE: To culture astrocytes of optic nerve and to establish the cell lines for further study of healing process after optic nerve trauma. METHODS: Optic nerve astrocytes of infantile and adults with sudden death were cultured by tissue inoculation or tissue digestion with 0.25% Trypsin and 0.06% EDTA. The second and fourth passage cells were stained with HE and anti-GFAP, S-100 protein, Vimentin, and CD34 antibodies. RESULTS: The trypsinized astrocytes of infantile optic nerve reached confluence in 7 days, but the astrocytes of adults weren't successfully cultured. The cultured cells were in polygonal shape with processes; the cytoplasm was abundant and pink; the cells had light-blue nuclei. These cells were positive in GFAP, S-100 protein and vimentin staining, and negative in CD34 staining. CONCLUSIONS: The results showed that astrocytes of infantile optic nerve can be successfully cultured and trypsinization is a better method than tissue inoculation. The culture of infantile astrocytes is easier than that of adult astrocytes. Immunohistochemistry were used to determine the source and type of those cultured cells. PMID- 12579650 TI - A clinical analysis of idiopathic orbital inflammatory pseudotumor. AB - PURPOSE: To observe the clinical findings and response to treatment in patients with a diagnosis of idiopathic orbital inflammatory pseudotumor. METHODS: 209 idiopathic orbital inflammatory pseudotumor cases seen between Jan 1, 1978 and Dec 31, 1999 in our hospital were evaluated retrospectively. RESULTS: Of the 209 cases, 118 were male and 91 were female; there were 90 in the right eye, 81 left eye and 38 both eyes. Patients age ranged from 4 to 80 years (mean 44.4). Proptosis (66%), palpable mass (65%), swollen eyelid (55%), increased orbital pressure (55%) and motility restriction (48%) were the five most common presenting signs. According to radiologic and surgical findings, focal mass within orbit was the most frequent subtype (43%), followed by lacrimal inflammatory pseudotumor (32%), diffuse orbital inflammation (10%), myositis (8%). Perineuritis (2%), periscleritis (2%), acute inflammation (2%) and eyelid pseudotumor (1%) were rare clinical findings. The response to treatment (with a mean follow-up of 1.5 years) showed that the cure rate was 40% and the effective rate 97% after combined management of surgical resection, systemic steroid and local low dose radiotherapy. CONCLUSIONS: Although recurrence of IOIP is common, the success rate of treatment for this group of patients is high. PMID- 12579651 TI - Contrast of the effect of alomide and sodium cromoglycate in the treatment of allergic eye diseases. AB - PURPOSE: To observe the clinical effect of alomide and sodium cromoglycate eyedrops in the treatment of atopic conjunctivitis, vernal conjunctivitis and giant papillary conjunctivitis. METHODS: Randomized control study was used. Alomide and sodium cromoglycate eyedrops used to treat two groups of patients for two weeks. The effects of the two groups were compared. RESULTS: With respect of comprehensive therapeutic effect, the effectiveness rate was 83.6% in alomide group, and 63.3% in sodium cromoglycate group, which was not significantly different statistically. With respect of itching alleviation, alomide is more effective than sodium cromoglycate. No adverse effects were found in both groups. CONCLUSION: Our study suggests that alomide is a safe and effective drug in the treatment of allergic conjunctivitis. PMID- 12579652 TI - The killing effect of photodynamic therapy using benzoporphyrin derivative on retinoblastoma cell line in vitro. AB - OBJECTIVE: To evaluate the killing effect of photodynamic therapy (PDT) using new photosensitizer benzoporphyrin derivative complexed with human low density lipoprotein on retinoblastoma (RB) cell line in vitro. METHODS: The experiment of photodynamic killing effect on RB cell line in vitro was performed by using benzoporphyrin derivative (BPD or Verteporfin) and monochromatic light at the wavelength around 690 nm. Seven BPD concentrations (2,500 ng/ml, 1,250 ng/ml, 625 ng/ml, 312.5 ng/ml, 156.25 ng/ml, 78.125 ng/ml, 39.0625 ng/ml) and three energy densities (1.2 J/cm2, 2.4 J/cm2 and 3.6 J/cm2) were applied. The damage of the tumor cells was evaluated by MTT assay 24 hours after PDT. The changes of the ultrastructure of RB cells were observed under eleceronic microscope 4 hours after PDT. RESULTS: There was a significant dose-response relationship between tumor cell damage and BPD concentration in the medium under light irradiation at the energy density of 1.2 J/cm2. At each BPD concentration, the inhibition rate increased with the rise of energy density. RB tumor cell necrosis was found widely under electronic microscope. CONCLUSIONS: This study suggested that RB cells are very sensitive to the PDT induced by BPD in vitro. RB tumor cell were directly killed by photodynamic effect induced by BPD. PMID- 12579653 TI - Microvascular changes in the retrobulbar optic nerve in idiopathic intracranial hypertension. AB - PURPOSE: To examine the microvascular changes in the retrobulbar optic nerve in idiopathic intracranial hypertension (PTC). METHODS: Both optic nerves from a 29 year-old man with a two year history of PTC were examined histologically and morphometrically. A semi-automated image analysis system and paraphenylenediamine (PPD) stain were employed to resolve sufficiently the microvascular images for counts and measurement. RESULTS: There were 150 vessels distributed in the optic nerves which revealed the following: The average lumen of the vessels in outer sectors were larger than those of the inner sector vessels (168.17 microns 2 vs. 46.99 microns 2; p = 0.0338; OD; and 251.96 microns 2 vs. 130.02 microns 2; p = 0.029; OS) while in the normal control optic nerve the outer and inner area lumens were reversed in size-differential, but this did not show a statistical difference. The thickness of the PTC optic nerve vessel walls in the outer sectors was also greater than that of the walls in the inner sectors (4.95 microns vs. 2.67 microns; p = 0.013; OD and 5.25 microns vs. 3.34 microns; p = 0.019; OS); the same measurements in the normal optic nerve showed a reversed ratio, which was opposite that of the experimental group and but not statistically different. CONCLUSION: This pattern of microvascular changes is consistent with the selective axonal loss in the peripheral area of each optic nerve as much more severe than that in the inner sectors. PMID- 12579654 TI - Immunocytochemical study of cells in the vitreous of proliferative vitreoretretinopathy. AB - PURPOSE: To identify the cellular components of vitreous samples obtained during vitrectomy for proliferative vitreoretinopathy (PVR). METHODS: With the use of three intermediate filament (IF) proteins, vimentin, glial fibrillary acidic protein (GFAP), and cytokeratin (CK), cytocentrifuge slides of 14 fresh vitreous aspirates were detected with immunohistochemical technique. RESULTS: All the specimens contained epithelial-like proliferative cells with or without pigment and some membrane-like pieces. Immunocytochemical staining showed that 76.0-90.0% cells stained for CK, 17.4-29.6% cells expressed GFAP, and 80.1-91.0% cells were positive for vimentin. CONCLUSIONS: Majority of cells in the vitreous samples originated from retinal pigment epithelial cells (RPE) and glial cells in PVR. Expression of IF proteins may be determined by tissue of origin and local microenvironment. PMID- 12579655 TI - Elevated vascular endothelial growth factor levels in the vitreous of patients with proliferative diabetic retinopathy. AB - OBJECTIVE: To detect the levels of vascular endothelial growth factor (VEGF) in the vitreous of patients with proliferative diabetic retinopathy (PDR) and to investigate the possible role of VEGF in the development of neovascularization in PDR. METHODS: Undiluted vitreous samples and fasting venous blood samples were obtained from 27 patients with PDR and 14 subjects with idiopathic macular hole who underwent pars plana vitrectomy. The concentration of VEGF was determined by quantitative enzyme-linked immunosorbent assay (ELISA). RESULTS: The level of vitreous VEGF in patients with PDR (median 0.41 ng/ml, range 0.09-11.56 ng/ml) was significantly elevated when compared with that in control subjects (median 0.017 ng/ml, range 0.008-0.04 ng/ml) (P < 0.001). The median of PDR patients' serum VEGF concentration was 0.19 ng/ml (0.090.46 ng/ml) which was far lower than vitreous VEGF concentration (P < 0.05). Vitreous VEGF concentration was higher in PDR patients with retinal detachment than that in patient without retinal detachment (P < 0.05). CONCLUSIONS: Vitreous levels of VEGF were remarkably elevated in eyes with PDR, and they were not influenced by its serum concentrations. VEGF played an important role in neovascularization of PDR. PMID- 12579656 TI - A rapid method of preparing the plasma membrane enriched fraction of the bovine retinal pigment epithelial cells. AB - PURPOSE: To devise a rapid method of isolating the plasma membrane enriched fraction (PMEF) of the bovine retinal pigment epithelial (RPE) cells with Percoll centrifugation medium. METHODS: The bovine RPE was homogenised with a tight fit Dounce homogeniser and centrifuged in a 16.7% Percoll gradient for 20 minutes. The RPE particulate fractions were characterised in terms of their protein concentrations, Na/K-ATPase and bicarbonate stimulated ATPase activities. RESULTS: The total protein recovery was 88.7% of the RPE homogenate. The nucleus layer was identified at the first band. The mitochondrial fraction was at the second layer according to its bicarbonate stimulated ATPase activity. The 3rd and 4th bands were enriched with plasma membranes and their Na/K-ATPase activities were 31.5 and 34.6 mumol/mg/h respectively. The Na/K-ATPase activities were about six times that of the RPE homogenate. CONCLUSIONS: A rapid method of isolating the bovine RPE PMEF has been devised which involved a single centrifugation procedure in a Percoll gradient. PMID- 12579657 TI - Effects of Tryptergium Wilfordii Polyglyco-sidium on the tumour necrosis factor levels in aqueous humor after intraocular lens implantation. AB - PURPOSE: Effects of Tryptergium Wilfordii Polyglyco-sidium (TWP) on the Tumour Necrosis Factor (TNF) levels in aqueous humor after intraocular lens (IOL) implantation were studied in rabbits. METHODS: Twenty-seven pigmented rabbits were divided into three groups. In the first group, the IOL were placed in the capsular bag after lens extraction. In the second group, rabbits received TWP therapy after IOL implantation. And in the third group, rabbit received prednisone therapy after IOL implantation. Aqueous humor samples were aspirated on the 1st, 3rd, 7th, and 14th postoperative day. A modified double antibodies indirect sandwich ELISA was used to detect for the presence of TNF. The data were closely studied by means of analysis of variance with SAS software. RESULTS: It was found that TNF level in aqueous humor reached its maximum on the 7th postoperative day in the group with IOL implantation. It was also found that the TNF levels in aqueous humor on the 7th and 14th postoperative day were significantly lower (P < 0.05) in the TWP-treated group with IOL implantation than those in the non-treatment group and the steroid-treated group. CONCLUSIONS: The current study offers evidence that TWP is more effective than the steroid in reducing the TNF level in aqueous humor after IOL implantation. PMID- 12579658 TI - Distribution of human lens crystallins and their sulphydryl contents of different age in two-dimension electrophoresis. AB - PURPOSE: To analyze water-soluble (WS) human lens proteins of fetus, adult and age-related cataract by two-dimensional IEF/SDS-PAGE electrophoresis. METHODS: DACM [N-(7-Dimethylamino-4-methyl-3-coumarinyl) maleimide] was used to determine the lens proteins sulphydryl (SH) content. RESULT: Protein SH contents in WS lens proteins have no significant difference among fetus, adult and age-related cataract lens. This is different from the relative published results obtained in lens proteins of animal cataract model using similar SH detecting methods. CONCLUSIONS: IEF/SDS-PAGE electrophoresis demonstrated that there were much more fragmentation of crystallins during lens development and cataractogenic process. It is suggested that this phenomenon is likely to be due to further conformational changes in the fragmented cyrstallins during aging and cataractogenic process. PMID- 12579659 TI - Clinical study of diclofenac sodium eyedrops used before and after cataract operation. AB - PURPOSE: To determine the effect of 0.1% diclofenac sodium (DS) eyedrops made in China on preventing surgically induced miosis and inflammation. METHODS: Seventy cases of cataract inpatients were randomly divided into two groups. DS eyedrops and normal saline as placebo were applied respectively 3 h, 1 h, 0.5 h before operation and once every morning for 7 days after operation. The pupil diameter was measured five times at different stages during the extracapsular cataract extraction. The eyepain, photophobia, conjunctival injection, KP, aqueous flare and light reaction of pupil were observed once a day after operation for a week. RESULTS: 37 eyes in DS group and 33 eyes in placebo group were included in the study. 32 of 37 eyes in DS group (86.5%) maintained mydriasis in all stages of operation and 33 eyes (89.2%) did not show any obvious sign of inflammation. There was statistically significant difference between the two groups. No serious side effects occurred in the DS group. CONCLUSIONS: The DS eyedrops can prevent surgically induced miosis. Its antiinflammatory effect was significantly better than that of placebo drops. PMID- 12579660 TI - Preliminary observation of one-site combined trabeculectomy and phacoemulsification in the management of cataract patients with drug-controllable glaucoma. AB - PURPOSE: To evaluate the effect of one-site combined phacoemulsification and trabeculectomy in the management of cataract patients with drug-controllable glaucoma. METHODS: Twenty-four cases (25 eyes) of cataract patients with glaucoma whose intraocular pressure could be controlled by drugs were observed. All had undergone one-site combined phacoemulsification and trabeculectomy. Intraocular pressure and visual acuity were observed. RESULTS: The follow-up was 1-21 months. Postoperative visual acuity was 0.4-0.6 in 9 eyes, 0.7-0.9 in 7 eyes and 1.0 or better in 9 eyes. Postoperative intraocular pressure was 2.17 +/- 0.764 kPa (16.28 +/- 5.73 mmHg), 1.83 +/- 0.90 kPa (13.74 +/- 6.77 mmHg) lower than that before surgery (t = 11.89, P < 0.01). Only one eye needed one drug to control intraocular pressure after surgery. CONCLUSIONS: One-site combined phacoemulsification and trabeculectomy is a safe, effective and convenient technique for the management of cataract patients with drug-controllable glaucoma. PMID- 12579661 TI - Loss of anterior chamber-associated immune deviation during normal pregnancy in primates. AB - PURPOSE: We recently found that loss of anterior chamber-associated immune deviation was associated with normal pregnancy in rabbits. The purpose of this study is to further investigate whether the same events occurred in nonhuman primates. METHODS: Mid-pregnant cynomolgus monkeys were randomly selected. Bovine serum albumin (BSA) was inoculated in anterior chamber of eyes of nonpregnant and mid-pregnant monkeys that were subsequently immunized with BSA in adjuvant. Delayed-type hypersensitivity (DTH) was assessed by skin challenge. RESULTS: Non pregnant monkeys of intracameral BSA proved able to acquire antigen-specific suppression of delayed-type hypersensitivity. By contrast, inoculation of BSA to anterior chamber of pregnant monkeys abolished the DTH-suppression effect. CONCLUSIONS: This is the first demonstration in primates that loss of anterior chamber-associated immune deviation occurred during normal pregnancy. The fluctuations of systemic hormone levels during normal pregnancy might influence local immunoregulation within the eye. PMID- 12579662 TI - The preliminary study of glucocorticoid receptor gene in Chinese patients with glucocorticoid-induced glaucoma. AB - BACKGROUND: To study the glucocorticoid receptor (GR) and the associated gene regulation in the pathogenesis of glucocorticoid-induced glaucoma (GIG) in Chinese patients. METHODS: The trabecular cells of normal individuals and patients with GIG were cultured in vitro. By using polymerase chain reaction (PCR), gene fragments on GR DNA binding sites of trabecular cells were amplified. The product was detected by gel electrophoresis. RESULTS: The trabecular cells were cultured successfully in normal individuals and patients with GIG in vitro. A single PCR product was obtained in both two groups with the same size of 545 base pairs. CONCLUSION: There is not any difference in gene on the GR DNA binding sites between normal individuals and patients with GIG. The results suggest the difference in mRNA or other functional genes. PMID- 12579663 TI - An experimental study on homoharringtonine liposome and glaucoma filtration surgery. AB - PURPOSE: To observe the inhibitory action of homoharringtonine liposome during the healing process of wounds in the filtering sites. METHODS: Posterior sclerectomies were performed in 14 rabbits. Postoperatively one eye of each rabbit received subconjunctival injections of HH liposome and fellow eye received saline injection in a randomized masked fashion. RESULTS: Fourteen days after operation the IOP of experimental eyes reduced significantly (P < 0.01) as compared with the controlled eyes, and the number of remaining filtering blebs increased noticeably (P < 0.05). Pathohistological examination revealed that the number of fibroblasts per square micron in the filtering sites and the thickness of the scars in the center of the filtering sites of the experimental eyes were less than those of the controlled eyes. No serious ocular toxic and side effects were found. CONCLUSION: This experiment suggest that homoharringtonine liposome can markedly inhibit the scar formation of filtering sites after glaucoma filtering surgery, promote the formation of filtering blebs, and provide an experimental evidence for its clinical use in future. PMID- 12579664 TI - Effect of liposome-encapsulated total alkaloid of harmaline on rabbit lens epithelial cells: experimental study on the prevention of posterior capsule opacification. AB - PURPOSE: To investigate whether liposome encapsulated total alkaloid of Harmaline (TAH) as a therapeutic agent is beneficial to prevention of posterior capsular opacification (PCO). METHODS: Liposome-encapsulated TAH was prepared by modified freeze-thawing method. 0.1 ml of liposome-encapsulated TAH (0.2 mg/ml) was injected into the capsular bag during extracapsular lens extraction (ECLE) of each eye in total 10 rabbit eyes. Blank liposome or balance salt solution (BSS) was used as control. Slit-lamp examination and histopathological examination was used to evaluated capsule opacification. Intraocular pressure (IOP), density and morphology of corneal endothelia cells, the amplitude and latency of b wave of ERG were measured. RESULTS: The inflammatory response was mild both in TAH treated and the control group. PCO formation occurred in the control group 2 weeks postoperatively, but the posterior capsule was clear in TAH treated eyes. 4 weeks and 8 weeks after operation, PCO occurred both in TAH treated and control eyes. However, it was milder in the TAH treated eyes. IOP remained at the normal level in all eyes. There was no difference in the density of corneal endothelial cells, and the amplitude and latency of b wave of ERG between TAH treated and control eyes. Histopathological study revealed that lens epithelial proliferation occurred 4 weeks and the Soemmerings ring developed 8 weeks postoperatively in the control eyes. However, only mild vacuolization and pkynotic changes of lens epithelial cells were found in TAH treated eyes. Transmmission electronic microscopy demonstrated that there were mild cytoplasm vacuolization and mitochondria swelling of lens epithelial cells in TAH treated eyes. CONCLUSION: The results suggest that liposome encapsulated TAH can inhibit metaplasia and proliferation of lens epithelial cells in the rabbit eye without obvious toxicity to the eye tissue. It may be used as a potential agent to prevent the development of PCO. PMID- 12579665 TI - A study on the compliance of the patients with ocular fundus diseases. AB - PURPOSE: To investigate the compliance of the patients with ocular fundus diseases with recommendation for follow-up examination after laser treatment, and the underlying reasons for non-compliance. METHODS: 53 patients with ocular fundus disease were asked to fill in a questionnaire which includes the socio demographic characteristics, fear of the fundus disease, conception of the laser treatment and the motivation for staying healthy. Variables were compared for the compliers group and the non-compliers group by chi-square test. RESULT: Of the 53 subjects, 35 were classified as non-compliers and 18 were classified as compliers. There was no statistically significant difference between the two groups on all selected socio-demographic factors, conception of the laser treatment and the motivation for staying healthy. Significant difference was found between the two groups on two of those items concerning the fear of the diseases (P < 0.05). About half of the participants showed the lack of knowledge about the laser treatment. CONCLUSION: The present study revealed the poor compliance of the patients with fundus disease. Patient's health beliefs influenced their compliance status. PMID- 12579666 TI - [Preliminary experimental study on commitment differentiation of embryonic stem cells induced by corneal limbal stroma in vitro]. AB - PURPOSE: To investigate the possibility of commitment differentiation of embryonic stem cells (ES cells) induced by corneal limbal stroma. METHODS: Superficial corneal limbal stroma and lentis epithelium from New Zealand white rabbit were used for feeder layer for ES-D3 cells culture to induce the ES cells differentiation into corneal limbal stem cells. The differentiated ES cells were determined by light microscope and electronic microscope. RESULTS: 1. The proliferation of differentiating ES cells which cultured on corneal surface stroma layer around the corneal limbus was slow. The appearance of induced ES cells were uniform cells with much larger size than undifferentiated ES cells, and the nucleoli become long and thin under electronic microscope, but the size of ES cells cultured on deep stroma maintain as small as undifferentiated ES cells. The ES cells cultivated on surface stroma layer around the corneal limbus appeared microvilli 2 weeks after grafting into subcutaneous of athymic mice. 2. Lentis epithelium can only delay but not induce the ES cells differentiation to corned limbal stem celluloid. CONCLUSION: These results suggest that the superficial corneal limbal stroma has a potential ability to induce ES cells commitment differentiation. The tertiary embryo induction may not occur in vitro. PMID- 12579667 TI - [Expression of vimentin in the lenses of vimentin transgenic mice]. AB - PURPOSE: To observe the expression of vimentin in the lenses of vimentin transgenic mice and investigate the role of vimentin in the formation of cataract. METHODS: The 12.7 kb chicken vimentin genes were microinjected into the male pronuclei of ICR mice. The injected embryos were transferred to the pseudopregnant recipient females and then generated several transgenic mice. Twenty mice developed cataract by cross-breeding. Expression of vimentin in 4 cataractous lenses and 4 normal lenses were detected by ABC immunohistochemical method. Usig SDS-PAGE and Western blot technique, we seperated and measured vimentin in the normal and two cataract lenses, in which one was part opacity and the other was complete opacity. RESULTS: Besides in the epithelial cells, vimentin was overexpressed in the cells in anterior cortex and equatorial region in the lenses of transgenic mice. By SDS-PAGE and Western blot, 57 KD urea soluble protein bands were seperated in normal and cataract lenses and were reacted with vimentin antiserum specifically. In normal lens, partly and completely opaque lenses respectively, the values of average grey density of these bands were 15.56, 34.59 and 33.06 and the percentages of vimentin in urea soluble fraction were 7.26%, 9.05% and 9.17%, according to the sum of the grey level values of each band. CONCLUSION: Over and ectopic expression of vimentin in the Lenses of transgenic mice inhibits the lens cell differentiation and is linked to cataract formation. PMID- 12579668 TI - [Genotype-phenotype correlation in Chinese patients with retinitis pigmentosa due to rhodopsin mutation]. AB - PURPOSE: To investigate the genotype-phenotype correlation in Chinese patients with retinitis pigmentosa caused by rhodopsin gene mutation. METHODS: On the basis of the onset of symptoms, degree of morphological changes and progression of visual disability in three (3/83) patients with identified mutations, the correlation of the phenotype with the corresponding mutations was assessed. RESULTS: There was a certain degree of allele-specificity. Severe form of retinitis pigmentosa was found in patients with mutation in the cytoplasmic domain and mild form of retinitis pigmentosa in patients with mutation in the intradiscal domain. CONCLUSION: Although there is a certain relation between the mutant rhodopsin and ocular manifestation, we need to accumulate more materials before relating a rhodopsin mutation to a specific phenotype. PMID- 12579669 TI - [Establishment of a chemotherapeutic agents-induced apoptosis model of retinoblastoma]. AB - OBJECTIVE: To determine the apoptotic effects of different chemotherapeutic agents on the retinoblastoma(RB) cell line HXO-RB44 and to establish a drug induced apoptosis model of RB in vitro as the basis for further research on the mechanism of drug-induced apoptosis and spontaneous regression of RB. METHODS: Twelve chemotherapeutic agents, including vincristine, cytarabin, methotrexate, cyclophosphamide, thiotepa, mitoxantrone, aclanomycin, pirarubicin, cisplatin, carboplatin, mitomycin, etoposide, of different concentration (10(-9), 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol.L-1) were employed into HXO-RB44 cell line respectively for 24 hours, then apoptotic effects on it were decided by observing the DNA ladders on agarose gel electrophoresis. After that, one chemotherapeutic agent with the most evident DNA ladders was applied into HXO-RB44 cell line for 4, 8, 16, 24, 48 and 72 hours respectively, and the DNA ladders on agarose gel were also surveyed. Apoptotic cells were identified with transmission electron microscopy. RESULTS: Typical DNA ladders were shown on agarose gel after HXO-RB44 cell line were exposed to 10(-6)-10(-5) mol.L-1 vincristine and 10(-5) mol.L-1 aclanomycin for 24 hours and the former were much clearer. No DNA ladders did emerge when the RB cells were treated by the rest ten chemotherapeutic agents. The DNA ladders began to appear when dealt with 10(-5) mol.L-1 vincristine for 8 hours, to be most obvious for 24 hours, and abated for 48 hours then disappeared for 72 hours. Amount of apoptotic RB cells were observed by transmission electron microscopy. CONCLUSION: Both vincristine and aclanomycin have the effect of inducing apoptosis on HXO-RB44 cell line, but that of vincristine is more forceful, which is time and concentration dependent. So vincritine is an ideal agent for establishing an apoptosis model of HXO-RB44 cell line. PMID- 12579670 TI - [The effect of erycibele alkaloid on intracellular Ca2+ mobilation of cultured human ciliary muscle cells]. AB - PURPOSE: To study the effect of erycibele alkaloid on cultured human ciliary muscle intracellular Ca2+ movement. METHODS: Intracellular Ca2+ was studied by Fura-2 probe and muscarinic antangonist was applied. RESULTS: Erycibele alkaloid had no any possible disturbulance on the examination system. The average resting[Ca2+]i was 79.3 +/- 28.7 nmol/Lol/L (n = 24). Erycibele alkaloid caused biphase [Ca2+]i elevation. Non-selective muscarinic antagonist Atropine (1 nmol/L) could completely block the 1 mumol/L erycibele alkaloid induced [Ca2+]i elevation; M1 antagonist Pirenzipine could not block the 1 mumol/L erycibele alkaloid induced [Ca2+]i elevation in low concentration (0.03 nmol/L) but in higher concentration (1 nmol/L). M3 antagonist 4-diphenyl acetoxy N-methy piperidine methiodide could completely block the 1 mumol/L erycibele alkaloid induced [Ca2+]i elevation in low and high concentration. CONCLUSION: The effect of erycibele alkaloid on increasing cultured human ciliary muscle cell intracellular Ca2+ is mediated by M3 receptor subtype. PMID- 12579671 TI - [The preliminary study of amniotic membrane on TGF-beta 1 expression after photorefractive keratectomy]. AB - OBJECTIVE: To investigate the effect of amniotic membrane on TGF-beta 1 expression after photorefractive keratectomy(PRK). METHODS: PRK was performed bilaterally on 6 New Zealand white rabbits. One eye was randomly tranplanted with preserved human amniotic membrane, and the other eye served as the control. The expression of TGF-beta 1 mRNA was detected with in situ hybridization at the 4th weeks after PRK. RESULTS: TGF-beta 1 mRNA expression was present in the corneal epithelium and stroma at the 4th weeks after PRK. The cornea in amniotic membrane transplanted group had also TGF-beta 1 mRNA expression, and the expression extent was less than that in control group (P < 0.05). CONCLUSION: Amniotic membrane can reduce the expression of TGF-beta 1 mRNA in cornea after PRK. PMID- 12579672 TI - [Observation of the endothelial healing in rabbit corneal alkali wounds by alizarin red S-trypan blue staining method]. AB - PURPOSE: To study the healing of the corneal endothelial cells in rabbit corneal alkali wounds. METHODS: 26 New Zealand white rabbits were used. A alkali wound was created with a round filter paper, 8 mm in diameter, which was soaked in 0.5 N NaOH for 5 seconds and was placed centrally on the cornea for 60 seconds. The rabbits were killed in 20 minutes; 1, 2, 6, 12 h; 1, 2, 3, 5, 7, 10, 14, 30 days after injury. The corneal endothelium was examined in the light microscope using a lizarin red S-trypan blue staining method. RESULTS: Twenty minutes after the wound, the corneal endothelial cells were destroyed. 72 hours following the wound, the endothelial cells surrounding the burns became larger, irregular and started to migrate onto the denuded descemet's membrane. The endothelial cells covered the burn with the spindle-shaped cells 7 days after wound. CONCLUSION: The endothelial damage is healed by the division and migration of remaining endothelial cells at the margin of the wound, which has some characters of fibroblast. PMID- 12579673 TI - [The role of spleen in induction and maintenance of anterior chamber-associated immune deviation in different species of animals]. AB - PURPOSE: The importance of the spleen to the induction of anterior chamber associated immune deviation (ACAID) in mice has been known for some time. Removal of the spleen within the first 4 days of injecting antigen into anterior chamber can abolish immune deviation and result in immunity. The purpose of this study is to explore the role of spleen in ACAID in other different species of animals. METHODS: After splenectomy and sham-splenectomy, bovine serum albumin (BSA) as a soluble antigen was inoculated into the anterior chamber (AC) of different animals (rat, rabbit and monkey) respectively. Recipient animals were immunized with BSA and complete Freund's adjuvant. Delayed-type hypersensitivity (DTH) was assessed by skin challenge. The maintenance time of deviant immune response was evaluated in the fixed interval. RESULTS: All the animals with receiving intraocular antigen inoculation, both in splenectomized group and in sham splenectomized group, did not display DTH reaction. In addition, the maintenance time of immune deviation in both groups after AC inoculation of antigen was not significantly different. In contrast, all the animals without receiving intraocular antigen inoculation, both in splenectomized group and in sham-sple nectomized group, displayed positive DTH reaction. CONCLUSIONS: The spleen may not play an important role in the induction and maintenance of ACAID in rat, rabbit and monkey. These data suggest that as the evolution progresses, the function of spleen may be replaced by other lymphatic organs. PMID- 12579674 TI - [The T cell activation from peripheral blood of patients with herpetic keratitis following in vitro activation]. AB - OBJECTIVE: To evaluate the T cells activation from peripherial blood(PB) of patients with herpes simplex keratitis following in vitro activation. METHODS: The flow cytometry was used to investigate the expression of HLA-DR, CD25, CD69, CD71 on T cell from PB, the PB in culture, the PB cultured with phytohemagglutinin(PHA) or phorbol 12, 13-dibutyrate(PDB) or PDB and ionophore (IONO), in 7 cases with herpetic stromal keratitis (HSK) and 4 cases with herpetic ulcer keratitis(HUK). 6 normal people were set as control. RESULTS: The percentages of CD3+DR71+ was higher, and the CD3+CD71+ was lower from PB of patients with HSK than those from normal people. After the PB was cultured for 72 hours, the percentages of CD8+CD71+ in lymphocytes from HSK and CD8+DR+, CD8+CD69+, CD8+CD71+ in lymphocytes from HUK were lower than those from nomal people. And the CD8+CD71+ cells from HUK also was lower than those from HSK. When the PB was cultured with PHA for 72 hours, the percentages of CD3+CD71+, CD8+CD69+, CD8+CD71+ from HSK and HUK were lower than those from normal people. The percentages of CD8+CD25+ in lymphocytes from HUK also was lower than those from normal, and the CD8+CD69+ cells in lymphocytes was lower than those from HSK. The percentages of CD3+CD71+ in PB cultured with PDB for 72 hours, CD3+CD69+ in PB cultured with PDB and IONO for 72 hours from HSK were lower than those from normal people. CONCLUSION: The ability of T cells activation decreases in cases with HSK following in vitro activation. The lymphocytes in patients with HUK shows much lower activation than those cases with HUK. PMID- 12579675 TI - [Activated T lymphocytes in epiretinal membranes from eyes of patients with proliferative diabetic retinopathy]. AB - PURPOSE: To investigate the potential contribution of immune-mediated processes to the development of proliferative diabetic retinopathy(PDR), an immunohistochemical study was undertaken to characterize the infiltrating immune cells in epiretinal membrances from the eyes of patients with PDR. METHODS: A total of 15 PDR epiretinal membrane specimens obtained surgically from pars plans vitrectomy were studied by using a panel of monoclonal antibodies against T lymphocytes (CD4), interleukin-2(IL-2) and interleukin-2 recpetors(IL-2R). RESULTS: Twelve of 15 specimens (80%) contained CD4-positive cells. IL-2 was found in 12 of 15 samples (80%), of which 11 also contained CD4-positive cells, and IL-2R was detected in 10 of 15 membranes(67%), of which 9 contained CD4 positive cells and released IL-2. Most of the IL-2R-positive membranes were from type I diabetic patients, 40% of them are younger than 40 years. CONCLUSION: Our study demonstrated the involvement of activated immune cells and release of lymphokine(s) in more than half of the diabetic epiretinal membranes tested and revealed that the processes of immune responses and the biological effects of lymphokines(s) may play an important part in the development of epiretinal membranes of PDR, especially in young-onset and type I diabetes. PMID- 12579676 TI - [Diagnostic value of B-scan in exudative age-related macular degeneration]. AB - PURPOSE: To explore the image features of exudative age-related macular degeneration (AMD) with B-scan. METHODS: Using NIDEK US-3300 model B-scan to observe 31 patients (54 eyes) with AMD. RESULTS: The membranous arc echo was found at the posterior polar and there was a shuttle absence of echo between the echo band and the wall of eyeball in the period of retinal pigment epithelium detachment. During the exudation and hemorrhage absorbed not comletely, mixed echo was seen. A small parenchyma body with protrusion was seen with stronger echo in the late phase when the exudation and hemorrhage were replaced by the scar tissues, it was like a convex lens or a triangle and the top pointed to the globe behind, but it was flat towards the vitreous. CONCLUSION: The B-scan examination is valuable in diagnosis, treatment and follow-up for exudative AMD. PMID- 12579677 TI - [The use of ultrasound biomicroscope in the diagnosis of anterior segment intraocular foreign bodies]. AB - PURPOSE: To evaluate the use of Ultrasound Biomicroscope(UBM) in the diagnosis of anterior segment intraocular foreign bodies(IOFBs). METHODS: Besides the rountine x-ray photography and B-scan ultrasound examination, 9 cases with difinite IOFBs injury history were examined by UBM. RESULTS: Nine cases of IOFBs were shown in UBM image, among them 4 cases were invisible in x-ray photography and B-scan ultrasound examination. The IOFBs located in anterior chamber(1 case), in posterior chamber (2 cases), in pars plana (5 cases) and in iris root(1 case). The size of IOFBs varied from 0.1 mm x 0.25 mm to 1 mm x 3 mm. The IOFBs were extracted by combined intraocular surgery or direct magnetize extraction. CONCLUSION: Although most IOFBs could be localized by X-ray and B-scan ultrasound, some located in the posterior chamber or ciliary body are hard to detect, especially when they are nonmetal and extremely small. The high resolution image of the anterior segment of the eye created by UBM can help to localize the IOFBs and show the relation between IOFBs and surrounding tissue, which is important in the diagnosis and the choice of surgery method. PMID- 12579678 TI - [Pathogenesis and clinical classification of the malignant glaucoma]. AB - PURPOSE: To study the pathogenesis of the malignant glaucoma and the basis for the clinical classification of malignant glaucoma. METHODS: Twelve eyes with malignant glaucoma were examed by ultrasound biomicroscopy (UBM), type A and type B ocular ultrasounographer and other clinical ophthalmic tests and diagnostic therapy. The ocular anatomic structures of all 12 eyes were compared with those of the fellow eyes. RESULTS: Ciliary block were confirmed in four of the twelve eyes (33.3%) with malignant glaucoma while the narrow and small anterior segment were found in the fellow eyes by UBM. Iris lens disphragm anterior displacement were found in the other eight of the twelve eyes (76.6%) and their ciliary bodies were deformed passively. The ciliary bodies and anterior segment of the fellow eyes of these eight eyes were anatomically normal. Diagnostic surgery was performed in seven of the twelve eyes and six cases with fluid existing in the vitreous cavity were identified. CONCLUSION: According to our study, we suggest to classify malignant glaucoma into two tyges. One is ciliary block, which caused by ciliary block and concerned with the narrow and small structure of the anterior segment of eye. The other is lens iris block, which caused by iris lens diaphragm anterior displacement. Both types have the same pathophysiologic characteristics, such as, shallow anterior chamber, posterior chamber disappearance and aqueous humor misdirection to the vitreous cavity. But their pathogengesis may not be completely same. PMID- 12579679 TI - [A preliminary report of a new technique: non-penetrating trabecular surgery]. AB - OBJECTIVE: To evaluate the clinical effect of nonpenetrating trabecular surgery with sodium hyaluronate gel implant in Primary Open-Angle Glaucoma(POAG). METHODS: Twenty-four eyes of 22 patients with medically uncontrolled POAG were underwent nonpenetrating trabecular surgery with hyaluronate gel implant. The procedure was performed involing excising a deep-sclera tissue that flush with innerwall of Schlemm' s canal without opening the anterior chamber, then placing a 3 mm x 4.5 mm x 0.5 mm or 3.5 mm x 3.5 mm x 3.5 mm sodium hyaluronate gel implant under the scleral flap respectively. The postoperative Intraocular Pressure (IOP), Inflammation and fitering bleb were analysised. All of patients were undertaken ultrasound biomicroscopy (UBM) to evaluate the surgery site on postoperative 1-3 months. Mean follow-up was 3.9 +/- 2.2 months. RESULTS: The IOP decreased from a mean preoperative value of 24.4 +/- 9.1 mmHg to a mean postoperative value of 15.1 +/- 3.7 mmHg (P < 0.01). The number of anti glaucomatous medications by topically or systemically was reduced from preoperative sorts of 2.8 +/- 1.4 to postoperative sorts of 0.7 +/- 1.0 (P < 0.01). The visual acuity remained stable, 6 eyes had IOP elevated and 2 eyes had occured hyphema. The complications such as flat chamber, inflammation and detachment of choroid were not observed. The UBM showed that the hyaluronate gel implant wasn't degraded and a transparent liquid space was formed under the scleral flap in all of patients on postoperatively 1-3 month. The filtering blebs were formed in some patients. CONCLUSION: Nonpenetrating trabecular surgery with hyaluronate gel implant can effectively lower IOP and reduce the number of anti glaucomatous medications. No severe complications occured. With a simple postoperative nursing, it can be performed in clinic. It is proved to be a promising and new special technique to treat glaucoma. PMID- 12579680 TI - [Expanded polytetrafluoroethylene with different pore diameter for keratoprosthesis cell ingrowth and corneal metabolism]. AB - PURPOSE: (1). To study the relationship between the different pore diameter of the expanded polytetrafluoroethylene, (ePTFE) and the rate of cellular ingrowth in cornea; (2). To evaluate the influence of implant on corneal metabolism. METHODS: Two kinds of ePTFE disk with 45 microns and 28 microns pore diameter were implanted after a free-hand intralamellar dissection. The eyes were examined by slit-lamp biomicroscopy up to 7 months postoperatively. The corneas were removed and examined by light and transmission electron microscopy. RESULTS: Stromal fibroblasts penetrate, proliferate, and synthesize collagen in the pore of these two kinds of ePTFE, while the rate of cellular ingrowth and collagen deposition was greater in the ePTFE with 45 microns pore than 28 microns pore. Lipid deposits occurred in 3 eyes with 45 microns pore and in 5 eyes with 28 microns pore (P > 0.05). Dicks remained in corneas for 7 months without any extrusion in all eyes. CONCLUSION: Stromal fibroblasts can penetrate, proliferate, and synthesize collagen in the pore of both two kind of ePTFE, while cells penetrate greatly in the larger pore. Dicks interfere corneal metabolism in some degree. The ePTFE with 45 microns pore diameter, 200 mm thick, 84% porosity is a suitable material for the skirt of the keratoprosthesis. PMID- 12579681 TI - [Clinical study on keratoplasty with corneal cryopreservation]. AB - PURPOSE: To evaluate the clinical application value of the corneal cryopreservation in keratonosus. METHODS: 32 patients with keratonosus treated by corneal transplantation were clinically observed. RESULTS: In 32 keratonosus cases, 59.37% (19 cases) of the grafts remained transparant; in 24 corneal ulcer preforation cases, 45.83% (11 cases) of the grafts were transparent, all eyeballs(100%) were remained. CONCLUSION: Corneal cryopreservation can be a practical, effective method in the treatment of keratonosus in district hospitals. PMID- 12579682 TI - [The improvement on related condition of corneal cryopreservation and roles of cryopreserved corneas in emergent keratoplasty]. AB - PURPOSE: To study the cryopreserved corneal endothelium viability on the condition that a series of related factors on long-term corneal cryopreservation were modified. The frozen corneas were used for emergent penetrating keratoplasty on poor condition patients. To evaluate the role of cryopreserved corneas on clinical application, especially on emergent keratoplasty. METHODS: Residual peripheral corneas were stained with Trypan blue and Alizarin red in order to assess endothelium survival rate after central grafts were punched. Specular microscope was used to measure corneal thickness and count endothelium density. The transparent rate of grafts, infective control rate, retinal restorative rate and the recovery degree of visual acurity were emphasized respectively. RESULTS: Endothelium survival rate of grafts was 82.1% averagely. Postoperative endothelium density was 1,642 cells/mm2 and graft thickness was 0.59 mm averagely. The epithelial defects healed after 2-5 days of the operation and edematous grafts were reclear after 2-4 weeks of the operation in all grafts, except for chemical burn and thermal burn. The transparent rate was 87.5%. Infective controll rate was 90.63%. Retinal restorative rate was 55.56%. Some useful visual acurity can be obtained when the infection was under controlled and the retinal was reattached. CONCLUSION: Modified long-term corneal cryopreservation is effective in maintaining endothelium viability and integrity. Furthermore, it could provide donors anytime because of it's unlimited preservative duration. Our study emphasizes especially the role of cryopreserved corneas in emergent keratoplasty. PMID- 12579683 TI - [Recent development of endoscope system in ophthalmology]. AB - The new ophthalmic laser microendoscope are widely used in a variety of clinical settings up-to-date. In this paper, we will describe the technical characteristics, recent development and prospectie evaluation of the microendoscope system. PMID- 12579684 TI - [Unfolded intraocular lens implantation through a 6 mm sutureless incision]. AB - OBJECTIVE: To evaluate the therapeuitic effects of phacoemusification and unfolded iutraoculavr lens (IOL) implantation through a 6 mm sutureless incision. METHODS: A 6 mm sclera tunnel incision was made at 12 o'clock tangent of the posterior limbus and unfolded polymethylmethacrylate(PMMA) IOL implantation was operated in 50 cases (55 eyes); 53 eyes were finished without suture after phacoemusification and 2 eyes were done with extra capsule cataract extration(ECCE) because of the broken posterior capsular. RESULTS: Postoperatively, the visual acuity was 0.6 or more in 92.4% of the eyes and 1.0 or more in 50.9% of the eyes at one week; 0.6 or more in 98.1% of the eyes and 1.0 or more in 67.9% of the eyes at one month. The mean postoperative astigmatism was 0.63 +/- 0.36 D at one week and 0.61 +/- 0.33 D at one month. There was no statistical difference between the mean preoperative astigmatism(0.60 +/- 0.48 D) and postoperative astigmatism at one week and one month (P > 0.05). CONCLUSION: The technique of phacoemusification and PMMA IOL implantation through a 6 mm sutureless incision can reduce early postoperative astigmatism, shorten the period of visual rehabilitation. PMID- 12579685 TI - [Subtenon's infiltration anesthesia for intraocular surgery]. AB - PURPOSE: To study the efficiency of Subtenon's anesthesia for intraocular surgery. METHODS: Subtenon's infiltration anesthesia was performed in the following ocular surgeries: Extracapsular cataract extraction combined with intraocular lens(IOL) implantation (14 eyes), cataract aspiration with IOL implantation(5 eyes), iridectomy(2 eyes), trabeculetomy(3 eyes), and removal of silicon band after scleral buckle for retinal detachment (1 eye). After superficial anesthesia with 0.5% Dicaine solution and incision of limbal conjunctira were performed, a 22 gauge blunt connected with a 5 ml syringe containing a mixture of 2% lidocain and 0.75% of Bupivacaine was introduced posteriorly along with the surface of the sclera towards both 10:00 and 2:00 o'clock direction, under Tehon's capsule at tabout 10-15 mm away from the limbus. A slow injection of 1.5-2.0 ml of the anesthesia was made at the equator of the globe. RESULTS: According to the standard made by Pang chenjiu, the efficiency of anesthesia was as follows: Grade I: 20 eyes(80%), Grade II: 4 eyes (16%), Grade III: 1 eye (4%). CONCLUSION: It is an effective, safe and simple anesthetic method for intraocular surgery especially for those with single eye, glaucoma eye in absolute stage and old patient with insufficient heart function. PMID- 12579686 TI - [Study on bacterial strains in conjunctival sac before and after intraocular surgery]. AB - PURPOSE: To investigate the changes of bacterial strains in the conjunctival sac before and after intraocular surgery, as well as clinical effects of preoperative antibiotic agents. METHODS: One hundred and thirty-three eyes with intraocular surgery included 68 eyes with cataract surgery, 39 eyes with glaucoma surgery, 20 eyes with retinal detachment surgery and 6 eyes with vitretomy. Bacterial culture and pharmacologic sensitivity test were performed on the conjunctival smear from the 133 eyes with intraocular surgery before and after the operation and the anterior chamber aspirate from the 68 eyes with cataract surgery after the operation. RESULTS: At the time of admission, 61 eyes(45.8%) of 133 eyes with intraocular surgery were culture-positive conjunctival smears with the following identified: Staphlococcus aureus: 23 eyes; Staphlococcus epidermidis: 25 eyes; Gram-positive rods: 13 eyes. 8 eyes(6%) had culture-positive conjunctival smears before disinfection, (Staphlococcus aureus: 3 eyes; Staphlococcus epidermidis: 5 eyes). One eye(1.5%) had culture-positive Staphylococcus epidermidis in anterior chamber aspirate of 68 eyes with cataract surgery at conclusion of surgery. Also, 4 eyes(3%) were culture-positive conjunctival smears after surgery. All the bacterial strains of the group were sensitive to Tobramycin, Gentamycin, Cephradinum(Velosel), Vancomycinum and Rifampin, but they were resistance to Tetracyclinum, Chloromycetin, Erythromycin and Ofloxacin. Eye Science 1999; 15: 267-269. CONCLUSION: In order to prevent bacterial endophthalmitis after intraocular surgery, installation of an useful antibiotic eye drop before the surgery is important. PMID- 12579687 TI - [Application of the treatment combined Western and traditional Chinese medicine in idiopathic uveo-encephalitis]. AB - PURPOSE: TO study the factors, which influence idiopathic uveo-encephalitis(IUE) in the occurrence, diagnosis, treatment and prognosis. METHODS: Analysing retrospectively 33 cases of IUE(66 eyes) treated with Combined Western and Traditional Chinese Medicine(CWTCM). The etiology, diagnosis and prognosis were also be discussed. RESULTS: Of the 33 cases, there were 21 cases having the history of eating Xin, Hot, fried food or certain special protein. 15 cases were misdiagnosed. With the therapy of CWTCM, 15 cases recovered completely, 12 cases improved in visual acuity and 6 cases showed no improvement or deterioration. CONCLUSION: Xin, Hot, fried food and certain special protein may interfere with the human immunity, which may be the cause of the IUE in certain patients. Fundus fluorescein angiography is helpful in early diagnosis, the method of CWTCM is a efficient treatment, which values for further investigation. PMID- 12579688 TI - Differentiation of embryonic stem cells into neurons and retina-like structure in nude mice. AB - PURPOSE: To investigate the intraocular growth and biological characteristics of mice embryonic stem cells in nude mice. METHODS: Murine embryonic stem cells (D3 cell line) were cultured and maintained in an undifferentiated state in vitro, then transplanted into the anterior chamber of nude mice. Mophological and immunohistochemical examinations were implemented. RESULTS: Two to three days after transplantation, yellow-white floating granules, sheets and masses were seen inside the anterior chamber and vitreous cavity, and enlarged gradually. 14 20 days later, the mice were executed. Morphological examination showed that there were undifferentiated cells and some round or polygonal differentiated cells in anterior chamber and vitreous cavity. The morphology of these differentiated cells were similar to that of the retina. The cells were highly positive in NSE staining. CONCLUSION: The transplanted embryonic stem cells could grow in the eyes of nude mice with tendency to differentiate into neurons and retina-like structure. PMID- 12579689 TI - Differential gene expression in retina of myopic chicken eyes using mRNA differential display. AB - PURPOSE: To study differentially expressed genes in retina of experimental myopic chicken. METHODS: Experimental myopia in chicken was induced by form-deprivation. The mRNA in chicken retina was analyzed by using differential display. RESULTS: Experimental myopia was successfully induced in chicken through form-deprivation. Differentially expressed gene fragments were detected in retina of chicken with myopic development and recovery as compared with normal controls. CONCLUSION: The differential display of mRNA may be a useful way in cloning myopic-related genes. PMID- 12579690 TI - The study of influencing factors on the growth characters of Sprague-Dawley rat retinal neurons in vitro. AB - PURPOSE: To investigate the influencing factors in culturing Sprague-Dawley (S-D) rats retinal neurons in order to lay foundation for further experimental research. MATERIALS AND METHODS: Retinal cells were plated on plastic plates and coverslips coated with poly-l-lysine or ethylene imine polymer for primary culture. The cultured cells were divided into following groups: 1. Culture medium changed every 2 to 3 days vs changed only once; 2. Cytosine arabinoside (Ara-C) added to the culture medium vs not added. The cells were observed and pictured under inverted phase contrast microscope. The cells were identified through immunocytochemistry. RESULTS: The immunofluorescence showed that most of the cultured cells were neurons, among them were a few retinal ganglion cells. In the cultured group of which substrata coated with poly-l-lysine and culture medium added with Ara-c, the neurons intended to aggregate into clusters with relatively straight neurites. In the group of which substrata coated with ethylene imine polymer and medium added with Ara-c, the neurons grew dispersively with bent neurites. Both of them survived for 2 to 3 weeks. The cells which plated in the medium not added with Ara-c did not aggregate into clusters and survived longer than 4 weeks. In the group of which medium changed several times, the survival time of neurons was shorter than that in the medium changed only once. CONCLUSIONS: The retinal neurons plated on the substrata coated with ethylene imine polymer are easy to observe because of its dispersive growth. It is not favorable for the growth of the neurons by changing culture medium many times. Ara-c may possibly have side effect on the growth of retinal neurons. PMID- 12579691 TI - Epidermal growth factor and its receptors family on the ocular surface. PMID- 12579692 TI - Corrective change of retinal thickness measured by optical coherence tomography and histologic studies. AB - PURPOSE: To evaluate the correlation of retinal thickness between optical coherence tomography (OCT) images and histologic slides. METHODS: Retinal thickness was measured in 16 rabbit retinal histologic slides. The same eyes had been previously measured by OCT for the comparison of results between two methods. Retinal thickness of each OCT image section was measured using both the manually assisted (requiring localization of reflectivity peaks by observer) and automated modes of the computer software. RESULTS: Retinal thickness measured by OCT demonstrated a high degree of correlation with retinal histologic study. The automated method (Cc = 0.66, P < 0.01) was less reliable than the manually assisted one (Cc = 0.84, P < 0.001). The former had an error in 95% confidence interval, ranged between -0.71 and 11.09 microns. The latter had a less error, ranged from -2.99 to 5.13 microns. CONCLUSION: OCT can quantitatively measure the retinal thickness. However, automatical identification of the reflective boundaries by computer may result in errors in some cases. To measure the retinal thickness by manually assisted mode can increase the accuracy. PMID- 12579693 TI - Characteristics of indocyanine green angiography in exudative age-related macular degeneration. AB - OBJECTIVE: To observe the characteristics of indocyanine green angiography (ICGA) in exudative age-related macular degeneration (AMD). METHODS: Thirty-one cases (39 eyes) were diagnosed as exudative AMD by ocular examination, fundus color photography, fundus fluorescein angiography (FFA) and ICGA. Their ages ranged from 50 to 82 years and visual acuities from FC/30 cm to 0.7. We analyzed and compared the characteristics of ICGA and FFA in these patients. RESULTS: Of 26 eyes with occult choroidal neovascularization (CNV) by FFA, 15 (57.7%) had classic CNV by ICGA. Of 4 eyes with serous retinal pigment epithelial detachment (PED) without CNV by FFA, 1 had serous PED with classic CNV by ICGA. The hyperfluorescence of the scar staining was detecfed by ICGA. CONCLUSION: ICGA adds clinically useful information and is an important adjunct in the evaluation, classification, and laser treatment of patients with occult CNV in AMD. PMID- 12579694 TI - New management of malignant glaucoma by phacoemulsification with posterior chamber foldable intraocular lens implantation. AB - OBJECTIVE: To investigate the feasibility of phacoemulsification with posterior chamber foldable intraocular lens implantation in the management of malignant glaucoma. PATIENTS AND METHODS: Fourteen patients with malignant glaucoma diagnosed in the Department of Glaucoma were enrolled in the study. 12 patients developed malignant glaucoma after filtration surgery. 1 developed after peripheral iridectomy. 1 patient developed malignant glaucoma without any clear cause. Cataract phacoemulsification was performed. Posterior chamber intraocular foldable lens was implanted in 10 cases of patients. RESULTS: All 14 patients were cured with normal intraocular pressure, normal anterior chamber depth and increased visual acuity. CONCLUSION: Phacoemulsification with posterior chamber foldable intraocular lens implantation is a good alternative in treating malignant glaucoma. PMID- 12579695 TI - Fresh amniotic membrane transplantation for conjunctival surface reconstruction. AB - PURPOSE: To determine whether fresh human amniotic membrane can be used to reconstruct the conjunctival detect created during symblepharon lysis. METHODS: Forty-two eyes of 39 consecutive patients with eye burns and Stevens-Johnson syndrome were randomized to accept fresh or preserved human amniotic membrane transplantation (AMT) during the period of severe scarring. Impression cytology was performed in 12 eyes with normal tear secretion which received fresh AMT. RESULTS: During a mean follow-up of 11 months (range, 6 to 18 months), thirty five patients (37 eyes) showed successful ocular surface reconstruction and resolution of motility restriction while four patients (2 eyes with fresh AMT, 3 eye with preserved AMT) with minimal recurrence of symblepharon. There was no significant difference statistically between two groups (Chi-square test). Amniotic epithelial cells can survive about three months after being transplanted onto ocular surfaces with normal tear secretion. CONCLUSION: Both fresh and preserved human amniotic membrane can be considered an ideal alternative substrate for conjunctival surface reconstruction during removal of severe symblepharon. PMID- 12579696 TI - Corneal topographical changes following strabismus surgery. AB - PURPOSE: To study corneal topographical changes after strabismus surgery. METHODS: Computer-aided corneal topography was used in 43 strabismus patients (45 eyes) one or two days prior to and six or seven days after strabismus surgery. The spherical and cylindrical equivalents were calculated based on the simulated keratometry. RESULTS: After the surgery, only the changes at 3 mm in the inferior quadrant were statistically significant. The changes at 3 mm in the rest quadrants and the changes at 7 mm were not significant. Significant changes in spherical equivalent were found post-operatively. Neither the horizontal nor the vertical meridional equivalent showed significant changes after surgery. CONCLUSIONS: The results of corneal topographical changes following strabismus surgery in our preliminary study indicated the little effect of strabismus surgery on corneal curvature and corneal astigmatism. PMID- 12579697 TI - Silicone oil tamponade combined with lensectomy preserving anterior lens capsule. AB - PURPOSE: To investigate the therapeutic effect and indication of silicone oil tamponade combined with lensectomy preserving anterior lens capsule. METHODS: Silicone oil tamponade combined with lensectomy preserving anterior lens capsule was performed in 33 cases (33 eyes) of high myopia, proliferative vitroretinopathy (PVR) D grade and giant retinal tear (GRT). 10 cases were only eye and 11 cases had harder nucleus of lens. The surgical methods included: 1. smashing lens nucleus and lensectomy preserving complete anterior capsule; 2. vitrectomy and membrane peeling; 3. usage of liquid perfluorocarbin or retinotomy for drainage; 4. silicone oil tamponade; 5. postoperative Nd; YAG laser for anterior capsulectomy. RESULTS: Follow-up time was 6 months or more in 29 cases. Total retinal reattachment was achieved in 22 cases, macular retinal reattachment in 5 cases. The visual acuity was 20/800 to 20/200 in 13 cases, 20/100 to 20/50 in 12 cases. Visual acuity was significantly improved in GRT group (P < 0.05). Complications included unexpected anterior capsule break intraoperatively, anterior capsule opacity, silicone oil emulsification and liquid perfluorocarbin remaining postoperatively. CONCLUSIONS: Silicone oil tamponade combined with lensectomy preserving anterior capsule was safe and can reduce the operative complications. The indications included: 1. GRT complicated with high myopia; 2. advanced PVR cases complicated with high myopia in which silicone oil must be used; 3. severe lens opacity cases in which silicone oil must be used; 4. complex retinal detachment of only eye. PMID- 12579699 TI - Modification of isolation and culture of human retinal pigment epithelial cells. AB - PURPOSE: To modify the isolation of human retinal pigment epithelial (RPE) cells and to increase the purification and production of cultured RPE cells. METHODS: The human eyecups were fixed on a rubber holder. After digestion by trypsin, RPE cells were collected, then cultured and identified by morphology, immunohistochemistry and electron microscopy. RESULTS: The cultured RPE cells grew actively in the early stage with transparent nucleus and abundant melanin particles in cytoplasm. These cells were positive in DOPA oxidase reaction and in anti-pancytokeratin antibody staining. Cellular microvilli and tight junctions could be seen through transmission electron microscopy. CONCLUSION: We developed a rubber holder to fix the eyecup. Using this holder, more and purer cultured RPE cells can be obtained. These cultured RPE cells are similar to those in vivo in morphology and immunohistochemical staining. PMID- 12579698 TI - Clinical investigation of radiation retinopathy fundus and fluorescein angiographic features. AB - PURPOSE: To investigate the fundus and fluorescein angiographic features in the patients with radiation retinopathy. CLINICAL MATERIALS: Color fundus photography and/or fluorescein angiography from 13 patients with nasopharyngeal carcinomas received external beam radiation were retrospectively analyzed. RESULTS: In this study, 26 damaged eyes of 13 patients developed some degree of radiation retinopathy. The earliest and most common finding was macular microvascular changes (microaneurysms and/or telangiectasia), which was observed in 100% (26/26) of the eyes. Intraretinal hemorrhages, macular capillary nonperfusion, and mascular edema were noted in 84%, 50%, and 42% of the eyes, respectively. CONCLUSIONS: Radiation retinopathy is common after external beam radiation of nasopharyngeal carcinomas. The prominent changes include macular microvascular changes, intraretinal hemorrhages and macular capillary nonperfusion. PMID- 12579700 TI - Rapid diagnosis of chlamydial conjunctivitis in laboratory. AB - PURPOSE: To evaluate the techniques of rapid and accurate diagnosis of chlamydial conjunctivitis. METHODS: Total 100 conjunctivitis patients (200 eyes) were studied. Twenty-two of 100 cases were diagnosed as chlamydial conjunctivitis. The infected epithelia were scraped from tarsal conjunctiva of both eyes and stained separately with Giemsa (100 cases) and immunofluorescence (anti-chlamydial antigen monoclonal antibody, 100 cases). RESULT: In immunofluorescent staining, 38 cases were seen positive staining and 62 were negative. In Giemsa staining, 29 were positive, and 71 were negative. In 22 cases with clinical diagnosis of chlamydial conjunctivitis, 13 cases were confirmed, and 9 were excluded by immunofluorescent staining. Technically, immunofluorescent and Giemsa stain takes 45 and 40 minutes, respectively. CONCLUSION: Comparing to Giemsa stain, 38 of 100 scraping specimens were positive (38%) by immunofluorescent staining, 29 of 100 per cent were positive by Giemsa staining. Giemsa staining takes 5 minutes less than immunofluorescent staining (40 versus 45 minutes), however, the positive staining in immunofluorescent staining is much easier to be recognized than Giemsa staining. PMID- 12579701 TI - [Effect of N-methyl-D-aspartate and dexamethasone on apoptosis of rat cultured retinal ganglion cells]. AB - PURPOSE: To investigate the effect of N-methyl-D-aspartate(NMDA) and Dexamethasone on cultured rat retinal ganglion cells (RGCS). METHODS: RGCs were obtained from 1-3 days old SD rats. In two groups, exposure of cocultured ganglion cells to N-methyl-D-aspartate (20-500 mumol/L) lasted for 24 hours. Apoptotic cells were identified by Hochest 33258 in one group; In another group, 0.4% Trypan blue dye was added. Cells excluding the dye were counted, and the survival rate of cells was determined by the ratio of the excluding cells in the experiment over in the control; Cocultured RGCs and purified RGCs were exposed to Dexamethasone(1 x 10(-4), 1 x 10(-5) and 1 x 10(-6) mol/L). After 24 hours, apoptotic cells were identified by Hochest33258. RESULT: Cocultured RGCs showed distinct morphological appearance of apoptotic cells when they were exposed to NMDA. The survival rate of cells was dose-related to the concentration of NMDA. Cocultured RGCs didn't show typical apoptotic appearance at 24 hours after exposure to Dexamethasone. But purified RGCs did so, even in the control group. CONCLUSION: NMDA has the effect of inducing cocultured RGCs apoptosis. Dexamethasone had no this effect on RGCs. Without additional neurotrophic factors, purified RGCs may become apoptotic cells when cultured in vitro for 24 hours. PMID- 12579703 TI - [The experimental investigation of epithelial healing in rabbit central corneal alkali wounds]. AB - PURPOSE: To observe the morphologic changes of epithelium in the alkali wounds at the central corneal of rabbits. METHODS: 38 rabbits weighing 2.5 kg were used. A alkali wound of the corneas was performed in the right eye of 38 New Zealand white rabbits with round filter paper, 8 mm in diameter, which were soaked in 0.5 mol/L NaOH for 5 seconds and then were placed centrally on the cornea for 60 seconds. The corneal burns were examined and photographed after the staining of 2% fluoresent staining with a slit lamp every day. The excised corneas were examined in the light microscope, electron microscope. RESULTS: The epithelial cells had rapidly disappeared in the burned area. The epithelial cells regrowth started at 8 hours following the burn, and 1-2 layers epithelial cells covered the burned area by 24 hours. The epithelial defects occured again on the 4th day. The desmosome and microvilli of the regenerating epithelium had a reduced number. There was no basement membrane between the regenerating epithelium and the strom. There were increased intercellular spaces between the regenerating epithalial cells. CONCLUSION: The epithelial healing were achieved through the division and migration of epithelial cells at the margin of the wound in rabbit central corneal alkali wound. The decrease of the junctions and the increase of intercellular spaces between the regenerating epithalial cells may be responsible for the second epithelial defects. PMID- 12579702 TI - [A quantitative study of the effect of daunoblastina on argyrophilic nucleolar organiser region-associated proteins in cultured human retinal pigment epithelial cells]. AB - PURPOSE: To investigate the effect of daunoblastina on argyrophilic nucleolar organiser region-associated proteins (AgNORs) in cultured human retinal pigment epithelium (RPE) cells. METHODS: The culture medium of secondary cultured human RPE cells were replaced with a daunoblastina-containing one at a dose of 180 micrograms/L, and the cells were incubated for 12 hr at 37 degrees C. After that, the medium was withdrawn, fresh medium was added and incubation was carried out for an additional 24 hr. AgNORs were studied by using a silver staining technique and image analysis system. RESULTS: The area of intranuclear AgNORs (micron 2/nucleus) in the control and the daunoblastina-treated group were 41.61 +/- 4.52 and 13.66 +/- 0.65(P < 0.01), and the area density of intranuclear AgNORs(AgNORs/nucleus area ratio)(%) in the two groups were 20.37 +/- 1.97 and 13.06 +/- 2.44(P < 0.01) respectively. CONCLUSION: Inhibiting nucleolar organiser region transcriptional activity may be partly responsible for the cytotoxicity of daunoblastina on the cultured human RPE cells. PMID- 12579704 TI - [Soluble IL-2 receptor levels in patients with Graves' ophthalmopathy]. AB - OBJECTIVE: Soluble interleukin-2 receptor (sIL-2R) levels in sera with Graves' ophthmopathy was investigated for its relationship to the clinical manifestations and therapeutic effects. METHODS: sIL-2R levels were measured in sera of 41 patients with Graves' ophthalmopathy which had not yet received specific treatment for their ophthalmopathy and euthyroid during the entire study period and 49 normal subjects. RESULTS: In untreated patients, the mean levels of sIL-2R were significantly higher than those in the normal subjects (P < 0.001). There was no difference in the levels of sIL-2R between patients with hyperthyriod Graves' ophthalmopathy and those with enthyroid Graves' ophthalmopathy. The levels of sIL-2R in patients with response for corticosteoids treatment were significantly higher than that in patients with non-response for corticosteoids treatment (P < 0.001), but there were no differences between the each class present in the NOSPECS classification of ophthalmopathy. 12 patients with response for corticosreoids had lower sIL-2R levels After 3-month treatment than pertreatment. CONCLUSION: These results suggested that elevated sIL-2R levels in patients with Graves' ophthalmopathy may be derived from ophthalmopathy itself, e.g. intra-orbital activated lymphocytes, and serve as a useful parameter to evaluate clinical activity score and predict value of corticosteoids treatment outcome. PMID- 12579705 TI - [Analyses of central serous chorioretinopathy from indocyanine green angiography]. AB - PURPOSE: To compare and analyse the characters of Indocyanine Green Angiography (ICGA) and Fundus Fluorescein Angiography (FFA) in Central Serous Chorioretinopathy (CSCR) and explore its pathological significance. METHOD: 35 cases of CSCR were examed with Heidelberg Retina Angiography. Simultaneous images of ICGA and FFA were analysed. RESULTS: Among the 35 cases, 29(83%) revealed more lesions in ICGA than in FFA. The lesions appeared in FFA were all associated with ICGA changes. CONCLUSION: The increasing of choroid hyperpermeability is an early change, which causes the overlying RPE dysfunction and structural damage. Some points related to the laser and medical treatment to CSCR were also discussed. PMID- 12579706 TI - [To evaluate the examinations both colour Doppler flow image and CT for orbital pseudotumors]. AB - PURPOSE: To explore the values of colour doppler flow image(CDFI) technique and computer tomographic(CT) scan in diagnosising orbital pseudotumors. METHODS: CDFI and CT were used for examining 67 cases of orbital pseudotumors demonstrated with surgery and pathaology. The results were treated with self-match design Chi square test. RESULTS: The diagnosis validities for 29 mass-type cases with CDFI and CT were 44.8% and 82.8%, respectively, with significant difference (chi 2 = 5.88, P < 0.05). The validities for 25 diffuse-type cases were 84.0% and 56.0%, respectively, with significant difference. (chi 2 = 4.0, P < 0.05). The validities for 19 mixture-type cases were 63.2% and 78.9%, respectively, with no significant difference (chi 2 = 1.63, P > 0.05). The diagnosis validities for all 67 cases were 63.0% and 72.7%, respectively, with no significant difference (chi 2 = 1.16, P > 0.05). CONCLUSION: The diagnosis validities with both imaging methods were not different for diagnosis of orbital pseudotumors. But the values for diagnosing various pseudotumors were different. The validity with CDFI was superior to CT in diffuse type; CT was superior to CDFI in mass type; while the validity with CDFI and CT were not significant different in mixture type. PMID- 12579707 TI - [Corneal limbal stem cell autograft and conjunctiva transplantation for treatment of pterygium]. AB - PURPOSE: To observe the therapeutic effects of corneal limbal stem cell autograft and conjunctiva transplantation for the treatment of pterygium. METHODS: Corneal limbal stem cell autograft and conjunctiva transplantation were performed on 38 cases (43 eyes) with newly developed or recurrent pterygium. The post-operative follow-up period ranged from 3 to 30 months (mean, 10 months). RESULTS: Of 38 cases (43 eyes), there were 34 cases (38 eyes) with stable epithelial healing and recovery of corneal transparency with no abnormal proliferation of pterygium-like tissue. 4 cases (5 eyes) were lost during the follow-up. CONCLUSION: To provide new stem cell source for injured limbus with limbal epithelial autograft transplantation is a reasonable therapeutic method for treatment of pterygium. PMID- 12579708 TI - [Cyclosporine A and thiotepa in prevention of postoperative recurrence of pterygium]. AB - PURPOSE: To evaluate the efficiency of Cyclosporine A and Thiotepa to decrease the postoperative recurrence rate of pterygium. METHODS: 50 patients (50 eyes) with pterygium were treated with bare sclera resection and divided into two groups. One group used 1% cyclosporine A eye drops, the other group used 0.05% thiotepa eye drops. RESULTS: The average follow-up time is 10 months. The recurrence rate of cyclosporine group and thiotepa group is 5% and 10%. CONCLUSION: Cyclosporine A and Thiotepa can inhibit the hyperplasia of pterygium, and decrease the postoperative recurrence rate. PMID- 12579709 TI - [Research progress of pathogenesis of pigmentary glaucoma]. PMID- 12579710 TI - [Calf trabecular cell culture in vitro, morphological and cytoskeletal ultrastructures and kinetics investigation]. AB - OBJECTIVE: To establish the method of culturing calf trabecular cells (CTCs) in vitro, to understand the morphology and function of CTCs, to probe into the effect of resistance of aqueous outflow in the pathogenesis and mechanism about primary open angle glaucoma(POAG). To direct the clinical using of drugs and to open up new antiglaucomatous medicine by pharmacological studies of CTCs. METHODS: Trabecular meshwork was collected from twenty eyeballs of calf donors after slaughter. The tiusse was primarily cultured and cells were subcultured. The growing characteritics and morphological features of cultured primary and passaged cells were observed by light and electron microscopes. Cell kinetics of the third and tenth passaged cells were analysed using autoradiography and flow cytometry. The influence of the antiglaucomatous drugs 0.25 mg.ml-1 epinephrine (EPI) and 0.025 mg.ml-1 dipivalyl epinephrine (DPE) on cell kinetics of the third passaged cells was studied. RESULTS: The growing characteritics and morphological features of cultured CTCs were as same as those of human trabecular cells. Growing types of CTCs included most of epitheial cell and few of fibroblast. The amount of cellular microfilaments was reduced, DNA synthesis time(Ts) and cell cycle time(Tc) were obviously prolonged with passaged increasing. Antiglaucomatous drugs-EPI (0.25 mg.ml-1) and DPE (0.025 mg.ml-1) made microfilaments dissolving, Ts and Tc obviously prolonging. CONCLUSION: Establishing the method of culturing CTCs in vitro and understanding their morphology, function and pharmacological effects provided an important information for studying human trabecular cells and probing into the effect of resistance of aqueous outflow in the pathogenesis and mechanism about POAG. These studies indicated that antiglaucomatous drugs-EPI (0.25 mg.ml-1) and DPE (0.025 mg.ml-1) influenced obviously microfilaments and cell kinetics of the third passaged cells and suggested that it is not to be ignored that 1% EPI and 0.1% DPE may make CTCs' microfilaments dissolving and may inhibit CTCs' division and proliferation when they are used in clinical therapy. PMID- 12579711 TI - [Intraobserver reproducibility study of parameters for measurement of position and height of ciliary process by ultrasound biomicroscopy]. AB - OBJECTIVE: To search for suitable measurement parameters for the study of the position and height of cilliary process by ultrasound biomicroscopy (UBM), to test the intraobserver measurement reproducibility of the new parameters we designed as well as the old ones. METHODS: Five parameters: TCPD, A angle, B angle, theta 4 angle and T value, are measured in 21 pieces of UBM image by one observer on five separate occasions. The intraobserver reproducibility is assessed by calculating the coefficient of variation (CV) and the difference among the parameters is tested by grouped T-test. RESULTS: Among 21 images, parameter A angle and T value have no image with CV > 10%, TCPD and B angle have 2 respectively and theta 4 angle has 5. As to average variation level, A angle is lower than TCPD, theta 4 angle is higher than TCPD, both differences of which are significant, and B angle and TCPD are about the same level, the difference between them is not significant. CONCLUSION: Among all the parameters reflecting the position of ciliary process in UBM image, A angle has the best intraobserver reliability, TCPD and B angle are less reliable, theta 4 angle has the worst reliability. The parameter reflecting the height of ciliary process, T value has very good intraobserver reliability. PMID- 12579712 TI - [Implantation of the molteno tube for refractory glaucoma following penetrating keratoplasty]. AB - PURPOSE: To assess the surgical effectiveness of the molteno tube implantation in glaucoma secondary to penetrating keratoplasty. METHODS: 9 eyes in critical condition with glaucoma secondary to corneal graft surgery, whose intraocular pressures (IOP) have previously failed to respond to glaucoma medication, were undergone Molteno tube implantation. RESULTS: IOPs ranged from 0.8-2.8 kPa (1 kPa = 7.5 mmHg) were obtained in 7 eyes, the success rate accounted for 7/9, 2 eyes still required installation of beta-blocker, the graft rejection occurred in one eye and was remedied with medical treatment, the graft remained half-transparent in one eye. Visual acuity improved in 4 eyes, stayed the same in 4 eyes and was worse in one eye. The best corrected visual acuity over 0.05 was recorded in 4 eyes. CONCLUSION: Moltemo tube implantation is a rational and effective method for preserving a certain level of visual acuity in patients with glaucoma following penetrating keratoplasty. PMID- 12579713 TI - [A comparison on postoperative shallow anterior chambers with different kinds of glaucoma surgery]. AB - PURPOSE: Comparing the rate of postoperative shallow anterior chambers with three kinds of filtration surgery to find the most effective way of reducing the complication. METHODS: The 120 cases (154 eyes) with primary angle-closure glaucoma or primary open-angle glaucoma were enrolled in this study. The patients were divided into three groups, group 1 (12 cases, 14 eyes) treated by deep sclecatony and group two (66 cases, 82 eyes) treated by trobeculecotomy and group three (42 cases, 58 eyes) treated by trobeculecotomy combining with scleral flaps tightly sutured with releasable sutures. RESULTS: The rate of shallow anterior chambers was following: group one 21.4%, group two 19.5%, group three 5.1%. The rate of this complication between group 1 and group 2 was not different in statistics (P > 0.05), but group 3 was significantly reduced in comparing with that of the other two groups (P < 0.05). CONCLUSION: The results indicate that this kind of suture may decrease the occurrence of the complication of shallow anterior chamber in filtration surgery and it is simple to operate and the curative result is reliable. PMID- 12579714 TI - [Result of phacoemulsification and intraocular lens implantation combined with trabeculectomy in the treatment of glaucoma with cataract]. AB - OBJECTIVE: The study was designed to investigate the application of Phacoemulsification and Intraocular Lens Implantation combined with Trabeculectomy (Phaco & IOL & T) for glaucoma with cataract. METHODS: "Phaco & IOL & T" was performed on 45 eyes of 45 patients in comparing with the treatment of Extra Capsule Cataract Extration and Intraocular Lens Implantation combined with Trabeculectomy (ECCE & IOL & T) on 40 eyes of 40 patients. The average period of follow-up was six months. RESULTS: The average Intra Ocular Pressure (IOP) in "Phaco & IOL & T" group was 2.08 kPa, astigmatism was 0.84 +/- 0.63 D and 84% of patients had the visual acuity > or = 1.0. The average IOP in "ECCE & IOL & T" group was 2.78 kPa, astigmatism was 1.54 +/- 0.67 D and 67.5% of patients had the visual acuity > or = 1.0 after three months of the operations. The difference was significant (P < 0.05). There was no severe complications in "Phaco & IOL & T" group while there was 9 cases with flat filtration pad in "ECCE & IOL & T" group. CONCLUSION: The method of "Phaco & IOL & T" is more efficient and safe than that of "ECCE & IOL & T". It should be the first consideration in the management of glaucoma with cataract. PMID- 12579716 TI - [Diathermic high-frequency capsulorhexis in cataract surgery]. AB - PURPOSE: To evaluate the efficiency of diathermic high-frequency capsulorhexis in cataract surgery. METHODS: After injection of viscoelastic material into the anterior chamber, a continuous circular capsulotomy (CCC) was performed using a diathermic high-frequency capsulorhexis machine in 34 eyes of 30 cataract patients during extracapsular cataract extraction (ECCE) combined with intraocular lens (IOL) implantation. RESULTS: Successfully CCC was made in 31 eyes. A small peripheral radial capsular tear was detected in 3 eyes. No postoperative corneal edema and other complications were found in all eyes. The postoperative corrected visual acuity > or = 0.5 was accounted for 91.2%. CONCLUSION: Diathermic high-frequency capsulorhexis is a simple, safe and effective method for ECCE (or Phacoemulsification) combined with IOL implantation. It is especially indicated for those eyes with hypermature cataract, traumatic cataract and children with congenital cataract. PMID- 12579715 TI - [The postoperative complications and management of excimer laser in situ keratomilleusis]. AB - OBJECTIVE: To evaluate the safety of excimer laser in situ keratomilleusis (LASLK) METHODS: The postoperative complications in 699 eyes of LASIK were observed and statistically analyzed. RESULTS: The haze, halo, growth of corneal epithelium under valve, the foreign body under valve, the fold of corneal valve, infection, decentralization, irregular astigmatism, valvular damage, low contrast sensibility, undercorrection and over correction, the psychological dysopia, glaucoma, fluctuating visual acuity and the loss of the best corrected visual acuity were found. CONCLUSION: LASIK has it's advantages of accurate manipulation, extensive range of correction and rapid recovery, but some of the complications resulting from this operation are so difficult to treat that careful consideration should be given before using this technique. PMID- 12579717 TI - [Management of infectious endophthalmitis following phacoemulsification]. AB - PURPOSE: To discuss the cause and management of infective endophthalmitis following phacoemulsification. METHODS: Anterior chamber irrigation with anterior chamber drug injection, postoperative systemic and topical antibiotic therapy as well as the use of topical mydriatic agent were applied to 5 of 3,850 eyes which showed endophthalmitis following phacoemulsification. RESULT: All the 5 eyes were saved from infective endophthalmitis. 1 to 5 eyes underwent vitrectomy, 4 of 5 eyes underwent anterior chamber irrigation (wash out). 4 of 5 eyes had satisfactory visual outcome. One of them had poor visual outcome. CONCLUSION: Preoperative contamination of the anterior chamber is the most common source for pathogenic agents responsible for endophthalmitis following phacoemulsification. Prompt anterior chamber irrigation (washout) with anterior chamber drug injection is one of the effective treatment. PMID- 12579718 TI - [A new anaesthetic method of intraocular surgery: anaesthesia at infrafascia bulbi]. AB - OBJECTIVE: To probe the advantage, indications and its mechanism of anaesthesia at intraocular operation with infrafaseia bulbi anaesthetic (IBA) technique. METHODS: 456 eyes were operated with the anaesthesia in infrafascia bulbi, including 77 eyes for phacoemulsification intraocular lens implantation, 346 eyes for modern surgical extracapsular cataract extraction and intraocular lens implantation, 33 eyes for trabeculectomy. 2 ml of mixture of 2% Lidocaine and 0.75% Bupivacaine were injected at infratemporal near bulbi conjunctiva of limbus through the blunt silicone canal, which entered the fascia bulbi via a small incision and reached the equator of the eyeball. When the injection of the mixture filled up the fascia bulbi the surgery could be performed. RESULTS: IBA applied in 456 eyes had shown the same effect as posterior fascia bulbar and peripheral fascia bulbar anaesthesia. IBA is a safe method. CONCLUSION: It is avoided the mechanical injure to optic nerve, artery, vein, muscle and wall of eyeball with IBA using blunt silicone injecting canal. It is proved in our clinical practice that this is a saft and efficient local anaesthetic method. PMID- 12579719 TI - [Analyses and management of 468 cases of eye trauma]. PMID- 12579720 TI - Preliminary study on in vitro induced differentiation of embryonic stem cells into neurons. AB - PURPOSE: To study preliminarily in vitro induced differentiation of embryonic stem cells into neurons for further investigation of an alternative for the treatment of glaumatous neuropathy. MATERIALS AND METHODS: Supernatant of cultured Buffalo rat liver cells (buffalorat liver cell-conditioned medium, BRL CM) was used for culturing embryonic stem cells (ES-D3 cell line). Morphological features of undifferentiated ES cells were studied by HE staining and electron microscopy. Based on the methods used by Bain et al, we modified the methods and used retinoic acid (RA) as an inducer to differentiate ES-D3 cells and cytosine arabinoside (Ara-C) as inhibitor of proliferative cells. The growth of the cells was observed under phase contrast microscope. RESULTS: ES-D3 cells cultured by BRL-CM grew in aggregates and remained undifferentiated. Electromicroscopy showed large nucleus and a large amount of mitochondria in undifferentiated ES cells and many processes on the surfaces. In the first day after the adding of retinoic acid, some neuron-like cells with one, two or more processes were present. In the second day after adding RA and the first day after the plus of 10 microns Ara-C, a large amount of neuron-like cells appeared, with the formation of neuron-like networks. CONCLUSIONS: Combined use of RA and Ara-C can induce ES cells to differentiate into neuron-like cells. Our present preliminary study might provide insights into an alternative for the treatment of glaucomatous neuropathy by the transplantation of embryonic stem cells. PMID- 12579721 TI - Quantitative study of basic fibroblast growth factor in vitreous with proliferative vitreoretinopathy. AB - OBJECTIVE: To quantitatively study basic fibroblast growth factor (bFGF) in the vitreous of proliferative vitreoretinopathy (PVR) in order to understand the role of bFGF in the development of PVR. METHOD: High sensitive sandwich enzyme immunoassay technique (ELISA) was used to measure bFGF level in vitreous of normal eyes, the eyes of PVR-C or PVR-D grade, eyes of vitreous hemorrhage and the serum levels of bFGF in PVR-D patients. RESULTS: The levels of bFGF in the vitreous were: median 5.20 ng/L, quartile 15.47 ng/L in 20 normal eyes; median 3.12 ng/L, quartile 10.48 ng/L in 35 PVR-C eyes; median 46.56 ng/L, quartile 113.96 ng/L in 26 PVR-D eyes; median 1.40 ng/L, quartile 6.25 ng/L in 25 vitreous hemorrhage eyes. The vitreous bFGF level in PVR-D group was significantly higher than that in the normal group, PVR-C group and vitreous hemorrhage group (P < 0.01). The mean of serum-bFGF level was 18.33 +/- 3.39 ng/L. The vitreous bFGF level of PVR-D group was significantly higher than serum-bFGF level (P < 0.01). And the vitreous-bFGF level in PVR-D group was significantly higher in larger retinal tear subgroup. CONCLUSION: The results suggested that bFGF is involved in the development of PVR. PMID- 12579722 TI - Regulation effect of vascular endothelial growth factor on human fetal choroid vascularization. AB - PURPOSE: To investigate the spatial and temporal regulation effect of vascular endothelial growth factor (VEGF) on human fetal choroid vascularization. METHODS: The eyeballs of 54 human fetuses from the 9th week to the 40th week due to accidental abortion were studied by immunohistochemically staining for the expression of VEGF and proliferation cell nuclear antigen (PCNA). RESULTS: (1) The distribution of VEGF expression in the retinal pigment epithelium (RPE) decreased with the increase of age, the peak of which was between the 9th and 14th week. (2) PCNA immunoreactivity was localized within choriocapillaris endothelium. The expression level decreased alone with fetus age. In this period the choriocapillaris endothelium kept proliferation, differentiation, canalization and remodelled to form the choroid vessels. (3) Statistically significant correlations were shown between the expression of VEGF in the PRE and that of PCNA in choriocapillaris endothelium (r = 0.933, P < 0.01). CONCLUSION: VEGF expression in RPE was positively involved in modulating human fetal choroid vascularization. PMID- 12579723 TI - Gene rearrangement analysis of orbital lymphoid infiltrating disorders. AB - PURPOSE: To determine whether the use of polymerase chain reaction for B-cell gene rearrangement in patients with orbital lymphoid infiltrate disorders could be useful in the diagnosis of lymphoma, especially, in differentiating benign lesion from malignant one. METHODS: In addition to clinical, pathological, and immunohistochemical evaluations, 48 cases of orbital lymphoid infiltrate disorders were examined for immunoglobulin heavy (IgH) gene rearrangement by means of PCR to amplify the FR3 region with formalin-fixed and paraffin-embedded tissues. RESULTS: Gene rearrangement in the third frame-work of the IgH region was detected in specimens obtained from 15 cases of malignant lymphoma, 4 of reactive lymphoid hyperplasia and 3 of orbital pseudotumor. All of these patients showed a discrete band (100 bp) which reflected monoclonal proliferation of B lymphocytes. 5 cases of malignant lymphoma, 6 of reactive lymphoid hyperplasia and 15 of orbital pseudotumor did not show a discrete band on PCR. CONCLUSIONS: The FR3 region gene rearrangement of Ig heavy in patients with orbital lymphoid infiltrate disorders may be an additional diagnostic tool in differentiating benign from malignant lymphoid diseases and in offering a useful adjunct for diagnosis in difficult or unclear cases. It is a reliable and practical method of gene diagnosis in orbital lymphoid infiltrate disorders and helps to identify the molecular mechanism of malignant lymphoma. PMID- 12579724 TI - New management of angle-closure glaucoma by phacoemulsification with foldable posterior chamber intraocular lens implantation. AB - OBJECTIVE: To investigate the management of angle-closure glaucoma by phacoemulsification with foldable posterior chamber intraocular lens (PC-IOL) implantation. DESIGN: Retrospective, noncontrolled interventional case series. PARTICIPANTS: In 36 eyes with angle-closure glaucoma (ACG), there were 18 eyes with primary acute angle-closure glaucoma (PACG), 14 eyes with primary chronic angle-closure glaucoma (PCCG), 3 eyes with secondary acute angle-closure glaucoma (SACG) and 1 eye with secondary chronic angle-closure glaucoma (SCCG). INTERVENTION: Phacoemulsification with posterior chamber intraocular lens implantation. MAIN OUTCOME MEASURES: Postoperative visual acuity, IOP, axial anterior chamber depth. RESULTS: After a mean postoperative follow-up time of 8.81 +/- 7.45 months, intraocular pressure was reduced from a preoperative mean of 23.81 +/- 17.84 mmHg to a postoperative mean of 12.54 +/- 4.73 mmHg (P = 0.001). Mean anterior chamber depth was 1.75 +/- 0.48 mm preoperatively and 2.29 +/- 0.38 mm postoperatively (P = 0.000). Best spectacle-corrected visual acuity in 36 eyes ranged from 0.01 to 0.7 (20/200 to 20/30) postoperatively, which was better than preoperative VA ranging from hand movement to 0.4 (20/50) (P = 0.000). CONCLUSION: Phacoemulsification with posterior chamber foldable intraocular lens implantation can be a good alternative in treating angle-closure glaucoma. PMID- 12579725 TI - Retrograde labeling of adult rat retinal ganglion cells with the flurogold. AB - PURPOSE: To study the densities and distribution of retinal ganglion cells(RGC) in adult rat retinae with flurogold(FG) labeling retogradely. METHODS: FG was injected to the superior colliculi(SC) and dorsal lateral geniculate nuclei (dLGN) in adult rats and the retinae were examined by fluorescence microscopy at various periods of time. RESULTS: FG-labelled RGC were observed in the retina as early as 3 days after application of FG. The labelled cells gradually increased in density, reached 95% of the maximal number on days 7 and the maximal number on days 30. The density of labelled cells was higher in the posterior pole than in the peripheral area. The fluorescence intensity in labelled cells maintained up to 60 days. CONCLUSION: The FG retrograde labeling method is reliable and effective for quantity of RGC. PMID- 12579726 TI - The effects of triptolide on HLA antigens expression of corneal epithelial cells induced by interferon-gamma in vitro. AB - OBJECTIVE: To observe the effects of immunosuppressants triptolide (TL) and cyclosporine A (CSA) on HLA antigens expression induced by interferon-gamma(INF gamma) in vitro. METHOD: By using an indirect immunofluorescent method and analysing with ACAS-570, the abnormal HLA antigen expression of cultured corneal epithelial cells was induced by INF-gamma. After incubation with one of the immunosuppressants (CSA, TL) for 72 hrs, the amount of HLA-A BC and HLA-DR antigens was measured. RESULT: There was no significant difference (P > 0.05) between the group with CSA and the positive control group without CSA. In contrast to CSA, TL dramatically inhibited INF-gamma induced expression of HLA antigens of corneal epithelial cells (P < 0.001), compared with the control group without TL. CONCLUSION: TL had direct inhibition on the expression of HLA-ABC and HLA-DR antigens induced by INF-gamma in vitro, while CSA had no obvious inhibition. PMID- 12579727 TI - A study of toxic effect of mitomycin C on cultured bovine trabecular meshwork cells. AB - PURPOSE: To explore the toxicity of Mitomycin C (MMC) on trabecular meshwork cells. METHODS: Bovine trabecular meshwork cells were cultured in vitro and exposed to MMC of different concentrations. The cellular morphology, ultrastructure, mortality and phagocytosis was studied with light microscopy, transmission electron microscopy and methods of Wright's stain, etc. RESULTS: It was found that the toxic effect of MMC on the cells was in a dose-dependent mode. 1 x 10(-2) and 1 x 10(-3) mg/ml of MMC caused a large part of cells dead, 1 x 10( 4) and 1 x 10(-5) mg/ml of the drug had remarkable killing effect on the cells. 1 x 10(-6) mg/ml of MMC had still a mild toxicity, while 1 x 10(-7) mg/ml of MMC had not any influence on cellular morphology, mortality, and phagocytosis, etc. The safe concentration on bovine trabecular meshwork cells was 1 x 10(-7) mg/ml and the LD50 was between 1 x 10(-3) and 1 x 10(-4) mg/ml. CONCLUSIONS: Refering to previous data, we conclude that conventional clinical-application of MMC might do harm to trabecular meshwork cells. PMID- 12579728 TI - Bifunctional effect of human IFN-gamma on cultured human fibroblasts from Tenon's capsule. AB - PURPOSE: To study the effect of human IFN-gamma on in vitro cultured human fibroblasts from Tenon's capsule. MATERIALS AND METHODS: The effect of different concentrations of human IFN-gamma and mitomycin-C (MMC), 5-fluorouracil (5-Fu) on cultured human Tenon's capsule fibroblasts (HTCF) was measured using a MTT [3-(4, 5-dimethylthiazo-2-yl)]-2, 5-diphenyltetrazolium bromide; Thiazolyl blue) colorimetric assay. The results were analyzed using ANOVA of the statistical package for social sciences (SPSS) 9.0 version. The difference was considered to be significant if P < 0.05. RESULTS: The effects of MMC and 5-Fu on the growth of HTCF were negative, while the effects of IFN-gamma on the growth of HTCF were both negative (10(2)-10(4) units/ml in two experiments) and positive (10(6), 10(5), 10 units/ml in two experiments). The inhibition rate of MMC ranged from 5.73% to 46.9%, which was similar to the inhibition rate of 5-Fu ranged from 12.49% to 38.92% (P = 0.351). The inhibition rate of IFN-gamma in two experiments was smaller than MMC and 5-Fu (P < 0.05). CONCLUSION: IFN-gamma has bifunctional effect (both enhancement and inhibition) on proliferation of cultured HTCF. The antiproliferative effect of IFN-gamma was weaker than MMC and 5-Fu. Further study has to be carried out to document the inhibition of scar formation of filtration bleb by IFN-gamma and the molecular mechanisms of its bifunctional effect on HTCF proliferation. PMID- 12579729 TI - The study on the expression of keratin proteins in pterygial epithelium. AB - PURPOSE: To evaluate the expression patterns of several keratin proteins in pterygial epithelium. METHODS: The proteins of K8, K10, K14, K16 and AE3 in the epithelium of 8 pterygia and 4 normal conjunctiva were detected using immunofluorescent staining. RESULTS: In pterygial epithelium, the staining patterns of 5 keratin proteins were different with normal conjunctiva. No staining of K8, K10 and K16 was detected in normal conjunctival epithelium, but in pterygial epithelium, K8 and K16 were present in full thickness and K10 was present in superficial cells. In normal conjunctiva, only base layer of epithelium showed positive staining with antibody of K14 and AE3 proteins, but they were present in the full thickness in pterygial epithelium and the antibody of AE3 protein stained the superficial cells heavily comparing with base cells. CONCLUSION: The abnormal expression of keratin proteins (K8, K10, K14, K16 and AE3) in pterygia indicates that the epithelial cells in pterygia are in the state of hyperproliferation and keratinization, which may be the cause of the abnormal tear function in patients with pterygia. PMID- 12579731 TI - Microsurgical treatment of Mooren's ulcer. AB - OBJECTIVE: To evaluate effect of microsurgery of lamellar keratoplasty (LKP) on Mooren's corneal ulcer. METHODS: The effect, postoperative astigmatism, postoperative vision, ulcer recurrence of postoperation, and surgical complications of two groups of consecutive Mooren's ulcer inpatients treated respectively by non-microsurgery of LKP and microsurgery of LKP were analyzed. RESULTS: There were significant differences of the effect, postoperative astigmatism, postoperative vision, ulcer recurrence of postoperation, and surgical complications between the two groups. The effect and postoperative vision of the microsurgery-treated group were better than those of the non microsurgery-treated group. The postoperative astigmatism, ulcer recurrence of postoperation, and surgical complications of the microsurgery-treated group were less than those of the nonmicrosurgery-treated group. CONCLUSION: Microsurgery of LKP of Mooren's corneal ulcer can greatly improve the ulcer healing and postoperative vision, and reduce the surgical complication and the ulcer recurrence of postoperation. PMID- 12579730 TI - The analysis of pedigree GZ (Guangzhou). 1 with primary open angle glaucoma. AB - PURPOSE: To analyze the hereditary modality of primary open-angle glaucoma in China. METHODS: The genetic form of Pedigree GZ. 1 was analyzed using Mandalian hereditary rules. RESULTS: Pedigree GZ. 1 had following characteristics: 1) The pedigree had four generations, and there existed POAG patients in each generation; 2) Each patient had a parent with POAG. If the parents didn't suffer from the disease, their children would not. 3) The incidence of POAG in the relatives of the patients was 1/2. In addition, The age of onset, intraocular pressure, fundus and prognosis was different from each other in the patients. CONCLUSIONS: 1) Pedigree GZ. 1 is inherited as an autosomal dominant trait. 2) There exists individual differences of clinical manifestations in POAG patients. PMID- 12579732 TI - High frequency color Doppler image of choroidal detachment. AB - PURPOSE: To study the Color Doppler Image (CDI) characteristics of choroidal detachment and the applied value of CDI. METHODS: Seventy-two cases (74 eyes) of choroidal detachment were studied retrospectively. RESULTS: The typical ultragraph of chroridal detachment displayed one or several smooth hemispherical or lobuler circular thick bands, with convex side toward vitreous cavity. Most of the choroidal detachments were located before the equator, a few of them were beyond the equator. CDI displayed blood flow singnal in the band. Pulse Doppler showed the frequency spectrum features of retinal detachment band were similar to those of central retinal vessels, whereas the frequency spectum features of choroidal detachment bend resembled those of ciliary artery in some cases of retinal detachment (RD) accompanied by choroidal detachment. CONCLUSION: CDI could make a correct and precise diagnosis of choroidal detachment. PMID- 12579733 TI - [Expression of bcl-2 gene in human trabecular cells induced by dexamethasone]. AB - PURPOSE: To investigate the influence of dexamethasone on expression of bcl-2 gene in human trabecular cells. METHODS: The trabecular specimens from human eyes were cultured in vitro and the cultured cells of the third generation on slides were immersed in culture solution containing 10(-7) M dexamethasone for 6 hours, 12 hours and 24 hours respectively. The expression of bcl-2 gene was studied with LSAB. RESULTS: The positive cells of the bcl-2 protein could be identified after dexamethasone effected for 6 hours and the positive cell increased with time going on. CONCLUSION: The expression of bcl-2 gene can be induced by dexamethasone, it may be related with glaucomatic pathogenesis. PMID- 12579734 TI - [Human trabeculer cells in culture and cell identification]. AB - PURPOSE: To establish the method of culturing human trabecular cells(HTCs) in vitro and to determine identifyng main points concerning cultured HTCs. METHODS: Trabecular meshwork, corneal and scleral tissue were collected from sixty-eight eyeballs of thirty-four human donors less than six years of age within 24 hours after death. The tissues were primarily cultured and the cells subcultured. Cultured cells were observed by light and electron microscopes. Fibronectin(FN), laminin(LN) and IV collagen (IV C) in extracellular matrix(ECM) of the cultured cells were immunohistocytochemically stained by S-P method. RESULTS: HTCs easily grew in the event of young donors and short-time drawing materials. Trabecular meshwork should be accurately separated. Techniques in primary culture, and time and proportion in subculture should be suitabled. Experimental studies should select the third to fifth passages. The growing characteristics, morphological features of cultured HTCs under light and electron microscopes differed from those of corneal and scleral cells adjoining them. Ultramicrostucture distinguished HTCs from corneal interstitial cells. ECM stained by S-P method distinguished HTCs from scleral fibroblast. CONCLUSIONS: These studies suggest that it is not difficult to culture HTCs in general culturing condition as long as the main points about culturing them are known-well. The identification of cultured HTCs must be combined with three aspects: the growing characteristics and morphological features of cultured cells under light and electron microscopes, and the immunohistochemical stained peculiarities of FN, LN and IV C in ECM of cultured cells. To establish the method of culturing HTCs in vitro and to determine identifyng main points concerning cultured HTCs become an essential condition and an important guarantee in order to investigate the pathogenesis and mechanism on primary open angle glaucoma(POAG) deeply. PMID- 12579735 TI - [Interferon-alpha 2b inhibits proliferation of human tenon capsule fibroblasts]. AB - PURPOSE: To evaluate the potential effect of interferon-alpha 2b(IFN alpha-2b) as antifibrotic treatment after glaucoma filtering surgery. METHODS: The MTT method was used to study the inhibitory effect of IFN alpha-2b on human Tenon capsule fibroblasts in vitro. The antimetabolites including 5-fluorouracil (5-FU), homoharringtonine(H.H) and mitomycin C(MMC) were also assessed. RESULTS: The IC50 values of IFN alpha-2b, 5-FU, H.H and MMC on fibroblasts at 48 hours were 60.578 mg/L, 5.447 mg/L, 0.531 mg/L, 1.329 mg/L respectively. CONCLUSION: The effect of IFN alpha-2b to inhibit proliferation of fibroblast was less than that of antimetabolites. Further investigation of other aspects of suppress(suppress) wound healing as a possible antifibrotic agent should be deserved. PMID- 12579736 TI - [The significance of combination of electrophysiology and automated perimetry tests on the early diagnosis of primary open angle glaucoma]. AB - OBJECTIVE: The study was designed to analyze the relationship among the three electrophysiological tests and visual fields in primary open angle glaucoma (POAG) in order to find some more sensitive and specific parameters in earlier and early diagnosis of patients with POAG. METHODS: Thirty-six(70 eyes) patients with POAG, 8(12 eyes) glaucoma suspects and 30 (60 eyes) normal subjects underwent pattern electroretinogram (PERG), oscillatory potentials (OPs), pattern visual evoked potentials (PVEP) and Humphery automated perimetry tests respectively. RESULTS: The abnormal rate of PERG was the highest among the three electrophysiological tests in the glaucoma suspects and those of PERG and automated perimetry results were higher than others after early stage. In the correlation analysis of all objects, the amplitude of PERG had negative correlated with IOP and visual indices, but no correlation with C/D and VA. OPs had negative correlated with C/D, and no correlation with visual fields indices, intraocular pressure (IOP) and visual acuity (VA). The latency of PVEP had positive correlation with visual fields indices and C/D, but no correlation with IOP. CONCLUSION: Statistical result showed that different parameters could be selected according to different appearance of POAG. In the earlier stage before visual field defects could be found, PERG would be a more sensitive indicator if IOP were high; once C/D was abnormal, OPs and PVEP would be the better signs even though IOP was normal. PERG and Humphery automated perimetry were very useful indicators after early stage. It was significant in combination of many visual function tests on earlier or early diagnosis of POAG. PMID- 12579737 TI - [Study on the dynamic examination of anterior chamber angle by ultrasound biomicroscopy]. AB - PURPOSE: To study the factors that influence ultrasound biomicroscopy(UBM) examination on anterior chamber angle, and explore a dynamic differentiating method. METHOD: To compare the UBM morphologic characters of the anterior chamber angle of the high-risk chamber angle and adhered angle groups under different illumination. RESULT: Under routine working condition, some eyes of the high-risk group (9/12) displayed bombing iris with extremely narrowly-opened angle; but 3/12 cases and the control group showed bombing iris and closed angle. Under bright circumstances, all angles of the high-risk group showed widely-opened, but the control group remained closed. CONCLUSION: Different illumination influences the UBM examination of the anterior chamber angle significantly, we should differentiate the angle morphologic characters under different illumination if it seems to be closed. Factors that affect the UBM examination were also discussed. PMID- 12579738 TI - [A quantitative measurement of retinal nerve fiber layer thickness by optical coherence tomography in normal Chinese people]. AB - PURPOSE: To measure retinal nerve fiber layer(RNFL) thickness in normal Chinese people for the use of early diagnosis in glaucomatous patients. METHODS: Optical Coherence Tomography(OCT) was used in 111 eyes of 77 Subjects to create cylindrical cross sections of the RNFL around the optic nerve head with scan diameter of 3.46 mm. The differences in RNFL thickness were analyzed with age, sex, either eye. RESULTS: We found a decrease in RNFL thickness with aging especially in the superior quadrant(P < 0.05), but there weren't significant difference between right and left eyes, neither the sex(P > 0.05). The Mean RNFL thickness of different ages(mean +/- s) was as following: the superior quadrant 128.00 +/- 10.31-149.00 +/- 13.17, the inferior quadrant 132.95 +/- 9.54-142.33 +/- 10.60, the overall mean RNFL thickness 109.00 +/- 4.72-114.28 +/- 7.98 microns. CONCLUSION: The preliminary study showed that neither sex nor either eye were associated with significant differences in RNFL thickness. It is necessary to define the normative value by age group on a larger scale. PMID- 12579740 TI - [Retinal light damage: I. The influences of light intensity and exposure duration at moderate and low intensities of cyclic light]. AB - PURPOSE: To study the influences of moderate- and low intensity of cyclic light exposure on the light damage of rat retina and its relationship with light intensity and exposure duration. METHODS: Twenty five 8-10 weeks old of female Sprague-Dawley rats were divided into 5 groups randomly. One group were as controls reared in dark-room, others were exposured to 12 hour light: 12 hour dark cyclic white xenon light from 3 to 28 days. The light intensities were 90 115 Lux(100 Lux), 400-650 Lux(500 Lux), 800-1,150 Lux(1,000 Lux), 1,400-1,650 Lux(1,500 Lux) separately. After various duration of exposure, the retina were evaluated with light microscopy, electron microscopy and morphometric study. RESULTS: Except for the intensity of 100Lux group, all of the other exposure intensities can induce the light damage of rat retina. The photoreceptor cells were affected most early. The higher of light intensity and the longer of exposure duration, the severer of the photoreceptor cell loss. In the severely damaged eye, RPE were also damaged. CONCLUSIONS: For the light damage of rat retina which is induced by moderate- or low intensity of cyclic light, the damage degree was both affected by light intensity and exposure duration. PMID- 12579739 TI - [Analysis of rhodopsin and peripherin/RDS genes in Chinese patients with retinitis pigmentosa]. AB - PURPOSE: To disclose the mutation of rhodopsin and peripherin/RDS genes among Chinese patients with retinitis pigmentosa as there was no identified mutation through sequencing reported in Chinese. METHODS: Genomic DNA was prepared from the peripheral lymphocytes. Gene fragments of the rhodopsin and peripherin/RDS genes were amplified by the polymerase chain reaction. The PCR products were analyzed by Heteroduplex-SSCP technique. PCR samples with aberrant migrational bands were identified through direct sequencing or cloning sequencing. RESULTS: Three different mutations in the rhodopsin gene were found in 3 of the 83 patients with retinitis pigmentosa(Va1104Phe, Lys311Glu, Pro347Leu). Two of the three mutations have not been reported before. One of the two (heterozygous, Va1104Phe) was found in an isolated patient and the other (homozygous, Lys311Glu) in a family with autosomal recessive retinitis pigmentosa. Mutation of peripherin/RDS gene was not found in the 83 patients. CONCLUSION: Mutation in the rhodopsin gene is the common cause in Chinese patients with retinitis pigmentosa, either autosomal dominant, recessive or sporadic. PMID- 12579742 TI - [An experimental study on limbal stem cells autograft transplantation with cryopreservation]. AB - PURPOSE: The viability of limbal stem cells with cryopreservation and the treatment effect with cryopreserved limbal autograft transplantation for ocular surface disorders (OSDs) of stem cells deficiency in rabbits. METHODS: Limbal grafts included 2 mm of peripheral cornea and 2 mm of conjunctiva and OSDs models of corneal stem cells deficiency were created in right eyes of 12 rabbits. The grafts were cryopreserved in -196 degrees C liquid nitrogen by program-controlled Bio-Freezer. The limbal autograft transplantations for ocular surface reconstruction were performed after cryopreserved 60 days and 30 days. RESULTS: The stem cells deficiency result in corneal epithelium delay healing. Seven of 12 eyes were healed post-operative 22-26 days with a mean time of 24 +/- 1 days and four eyes with recurrent erosion. The swelling and neovascularization appearing with a mean time of 5 +/- 1 days were discovered. The cryopreserved limbal autograft transplantation can increase corneal epithelial healing with a mean time of 10 +/- 2 days. It was observed that corneal stromal swelling and neovascularization were markedly decreased, and neovascularization of two eyes were disappeared. CONCLUSIONS: Cryopreservation in liquid nitrogen can preserve the viability of limbal stem cells, and provide viable limbal grafts to treat OSDs in clinical patients. PMID- 12579743 TI - [The penetrating keratoplasty for the pseudophakic bullous keratopathy]. AB - PURPOSE: To investigate the clinical effectiveness of penetrating keratoplasty for pseudophakic bullous keratopathy and the indication for keeping or removing intraocular lens. METHODS: 13 cases(13 eyes) of pseudophakic bullous keratopathy were treated by penetrating keratoplasty. RESULTS: The intraocular lens were kept in 11 eyes and were removed in 2 eyes during operation, The average follow-up time was 16 months. All corneal grafts remain transparent, Visual acuity of most cases were improved, The best-corrected visual acuity of 5 eyes was 0.2 or better. CONCLUSION: The penetrating keratoplasty is an effective treatment for the pseudophakic bullous keratopathy. It should be kept if intraocular lens is in normal position and located in lens capsule, and it is helpful for safety of the surgery and improvement of visual acuity. PMID- 12579741 TI - [An investigation on chromosome aberration of SO-Rb50 cloned cell strains]. AB - PURPOSE: To compare the differences of chromosome aberration among 3 cloning cells (MC2, MC3 and MC4) of SO-Rb50 cell line. METHOD: G-banding and karyotype analysis were performed on the 11th passage cells of MC2, MC3 and MC4 cell strains. RESULTS: Both numerical abnormal and structural aberrations of chromosomes could be observed in these 3 cloned cell strains. Diploid cells had the preponderance over the aneuploid cells. However, some cells showed pseudiploid karyotypes. Several kinds of structural chromosome aberrations were observed. The chromosome aberrations in the same passage of different cell strains were different. Aberration of chromosome 13 was rare and the aberration feature were different in 3 cloned cell strains. Five marker chromosomes were identified. M1, t(1;1) qterp35::q24-ter could be found in all cell strains. Other markers were seen in different cell strains respectively. In our experiment, two marker chromosome in chromosome 2, M4 and M5, were found in SO-Rb50 cell line for the first time. CONCLUSION: Chromosome aberrations were different with different cloned cell strains. Combined with formerly study in SO-Rb50 cell line, we found the chromosome aberration had dynamic changes during the long-term culture in vitro. We suggest that aberration of chromosome 13 is not the only cause of Rb; aberration of chromosome 1, a common event in some neoplasias as well as in SO Rb50, plays a meaningful role in the immortalization of this cell line; other aberrations observed in our experiment could be considered as evidences of genomic instability of the cell strains and presumably enhance the growth potential of the cells in which they develop. PMID- 12579744 TI - [Morphological observation of the endogenous endophthalmitis of cryptococcus neoformans]. AB - PURPOSE: To observe the pathologic change of the endogenous endophthalmitis of cryptococcus neoformans. METHODS: Light and electron microscope were used. RESULT: The choroid was focally tumor-like by an inflammatory cells infiltrate, which consisted of multinucleated gicant, epithelioid cells and lymphocytes. Special stains (Grocott's methnamine silver, PAS, alcian blue) and electron microscope disclosed the budding form of cryptococcus neoformans. CONCLUSION: The morphologic examinations are only an effectivel method of diagrosing the endogenous endophthalmitis of cryptococcus neoformans. PMID- 12579745 TI - [Preparation of liposome encapsulated daunorbicine and determination of daunorubicine of aqueous humor in rabbit eyes]. AB - PURPOSE: To investigate the method of determination the concentration of daunorubicine(DNR) and liposome encapsulated daunorubicine(LDNR) of aqueous humor in rabbit eyes. METHODS: After extracapsular lens extraction, 0.1 ml of 1 mg/ml DNA and LDNR were injected into the anterior chamber. At the 12, 24, 48 hours and 1 week after operation, the aqueous humor were aspirated to determine the concentration of DNR by high performance liquid chromatography(HPLC). RESULTS: At the 12, 24, 48 hours after operation, DNR were determined in all eyes, but at one week after operation, the DNR could only be detected in LDNR group. The concentration of DNR in LDNR group were higher than DNR group. PMID- 12579746 TI - [Observation of the effect of pediatric secondary posterior-chamber intraocular lens implantation]. AB - PURPOSE: To evaluate the effects of secondary posterior chamber lens implantation for aphakia in children. METHODS: Thirty-six consecutive pediatric aphakic eyes were retrospected. All eyes were secondarily implanted posterior-chamber lens using different operative techniques according to the condition of lens capsule remained after dissecting the posterior adhesion, forming extent space in posterior chamber. Follow-up ranged from 1 to 20 months. The effect of corrected visual acuity was observed. RESULTS: Postoperative nated visual acuity was equal or better than corrected visual acuity preoperatively in 22 eyes(61.1%). Postoperative corrected visual acuity was better than that before surgery in 32 eyes (88.8%) and was equal to that before surgery in 4 eyes(11.1%). CONCLUSIONS: Secondary posterior chamber IOLs implantation was an effective surgery for aphakic eyes in children, but attention should be paid to anti-inflammatory treatment and amblyopia treatment. PMID- 12579747 TI - [A preliminary report of the effect of levodopa and carbidopa for childhood amblyopia]. AB - PURPOSE: To explore the short-term effect of levodopa and carbidopa in 36 children (4.5-14 years) with refractory amblyopia. METHODS: 36 cases of incurable childhood amblyopia treated with occlussion with visual acuity stably for 6 months were retreated by levodopa and carbidopa 1.5 mg/kg, 0.38 mg/kg respectively for 3 months. Visual acuity, fusional and stereoscopic function, retinal light sensitivity and scotomas were observed at 1 and 3 months after tablet taken. Medicine side effects were observed. RESULTS: At 3 months, the visual acuity of 88.89% eyes were improved(average 2.27 +/- 1.26 line), fusional and stereoscopic function were restored in 10 and 6 cases respectively. Retinal light sensitivity significantly increased, scotomas disappeared in 19 eyes and contracted in 6 eyes. The basic cure rate was 42.86% Less side effect were found. CONCLUSION: Average dose levels(1.5/0.38 mg/kg) of levodopa and carbidopa were an effective and secuity method to improve visual function of refractory childhood amblyopia. PMID- 12579748 TI - [The clinical application of topical aneathesia in phacoemulsification]. AB - PURPOSE: To evaluate the effects of topical aneathesia in phacoemulsification. METHODS: 82 patients(91 eyes) with cataract in phacoemulsification were divided into two groups, topical aneathesia was used on the group of 40 patients(45 eyes); Regular local aneathesia was used on the other group of 42 patients(46 eyes). RESULTS: The preparatory time prior to operation in the topical aneathesia group was shorter than that in the regular local aneathesia group(P < 0.05). Eye sight recovery of topical aneathesia groups was quicker than that of the other group (P < 0.01). There was no distinct difference of operation time and pain killing effects between the 2 groups(P > 0.05). COUCLUSIONS: Topical aneathesia, safe, effective and easy to control, can be popularized in phacoemulsification. PMID- 12579749 TI - [Clinical research of trabeculectomy on outpatients]. AB - PURPOSE: To investigate the feasibility of Trabeculectomy on outpatients. METHODS: Postspectively compared and discussed the surgery effect and complications of trabeculectomy on 38 eyes of 27 outpatients with those on 38 eyes of 30 inpatients. All the surgeries were performed by the same surgeon in the same period. RESULT: The IOP of outpatients' group after one week, one month, three months or more are respectively 1.4800 +/- 0.5977 kPa, 1.8379 +/- 0.5517 kPa, 1.9650 +/- 0.4879 kPa, while the inpatients' group are 1.3468 +/- 0.5994 kPa, 1.7737 +/- 0.5177 kPa, 2.0144 +/- 0.5447 kPa. t-test has shown that the differences of the IOP after the surgery, the surgery effect and complications of trabeculectomy between the outpatients and the inpatients had no significance. CONCLUSION: Trabeculectomy on outpatients in hospital with satisfied facility would be effective, safe and feasible. PMID- 12579750 TI - [Clinical analysis of 2,088 cases of ophthalmic consultation in a general hospital]. PMID- 12579751 TI - [Clinical observation of intrachamberal anesthesia]. AB - PURPOSE: To observe the effect in operation and influence after operation of intrachamberal anesthesia. METHODS: 2 Percent lidocaine used for intravenous injection was diluted by Bss-plus to be 0.5 percent solution. Intraocular surgery of 63 cases(76 eyes) were performed intrachamberal anesthesia by anterior chamber puncture. RESULT: All of the operations were done well, the patients didn't feel painful. Complications were not found by following up ten months. CONCLUSION: This kind of anesthesial technique was regarded to be simple, safe, and efficient. It was worthy of searching in anterior intraocular surgery. PMID- 12579752 TI - [High throughput screening method of nitric oxide synthase inhibitors and enhancers]. AB - AIM: In order to discover new inhibitors and enhancers of nitric oxide synthase (NOS), an in vitro assay to determine NOS activity was established for high throughput screening. METHODS: The activity of NOS was detected based on the change of nicotinamide-adenine dinucleotide phosphate (NADPH) concentration in the reaction system by the fluorescence density. The enzyme was prepared from bovine brain by gradient centrifugation. The reaction performed in black 96 well micro-plate with a final volume of 90 microL. Every factor which would affect the results such as the concentration of NADPH, L-arginine (L-Arg, used as substrate) and enzyme protein was optimized in different conditions. At last, 5,600 samples (compounds and extracts) were screened by the method. RESULTS: The test signal (fluorescence density) in the reaction system was influenced by many different factors such as temperature and concentration of substrates. The ideal system contains protein 1.50 mg.mL-1, L-Arg 1 mmol.L-1, NADPH 0.1 mmol.L-1 at 37 degrees C. In this method, there were about 2% samples which emit fluorescence, and about 0.5% samples which quench the fluorescence. So these samples were deleted from the sample library. The effects of these samples on activity of NOS were distributed in a normal manner. About 2% samples had potential effects on the NOS activity (including inhibitors and enhancers). CONCLUSION: The method can be performed by high throughput screening and gives the stable data, not only for inhibitors, but also for enhancers of NOS activity. PMID- 12579753 TI - [Inhibitory effect and kinetic analysis of sodium quercetin-7,4'-disulphate on recombinant human protein kinase CK2 holoenzyme]. AB - AIM: To study the direct effect and kinetics of sodium quercetin-7,4'-disulphate (SQDS) on recombinant human protein kinase CK2 holoenzyme. METHODS: The recombinant human CK2 holoenzyme activity was assayed by detecting incorporation of 32P of [gamma-32P] ATP into the substrate in various conditions. RESULTS: The recombinant human CK2 was a second messenger (Ca2+, cAMP and cGMP) independent protein kinase. The characterization and function of the reconstituted holoenzyme were consistent with those of native CK2. SQDS was shown to strongly inhibit the holoenzyme activity of recombinant human protein kinase CK2 with an IC50 of 4.4 mumol.L-1, which was more effective than DRB and A3, known CK2 special inhibitors. Kinetic studies of SQDS on recombinant human CK2 showed: the inhibition was competitive with ATP and noncompetitive with casein. CONCLUSION: SQDS is a potent inhibitor of protein kinase CK2. This study provide experimental basis for the development of more effective inhibitors of CK2 and for clinical application of SQDS in the future. PMID- 12579754 TI - [Stereoselectivity in O-demethylation of trans tramadol in rat liver microsomes in vitro]. AB - AIM: To study the stereoselectivity in O-demethylation of trans tramadol. METHODS: With or without quinine and quinidine as inhibitors, rat liver microsomes were incubated in vitro with the enantiomers or the racemate of trans tramadol. The concentrations of the enantiomers of trans tramadol and O demethyltramadol in the incubates were determined by high performance capillary electrophoresis. The O-demethylation processes were assayed by using the enzyme kinetic analysis method. RESULTS: After incubation, the concentrations of (-)-O demethyltramadol were higher than those of (+)-enantiomer in all rat liver microsomal incubates. Enzyme kinetic analysis showed that the Km of the formation of the enantiomers of O-demethyltramadol were similar; The Vmax and Clint of the formation of (-)-O-demethyltramadol were significantly higher than those of the formation of (+)-enantiomer. When the racemate of trans tramadol was used as the substrate, there was interaction between the two enantiomers. The Km of the formation of the enantiomers of O-demethyltramadol increased, the Vmax of the formation of (+)-O-demethyltramadol decreased, the Vmax of the formation of (-)-O demethyltramadol increased slightly. The O-demethylation of the enantiomers of trans tramadol was shown to be inhibited competitively by quinine and quinidine. The Ki of quinine and quinidine were 1.6 and 10.8 mumol.L-1 to the formation of ( )-O-demethyltramadol, 0.8 and 3.4 mumol.L-1 to the formation of (+)-O demethyltramadol, respectively. Furthermore, quinine and quinidine were found to have stereoselective inhibition on the formation of O-demethyltramadol, both mainly inhibited the formation of (+)-O-demethyltramadol. CONCLUSION: The O demethylation of trans tramadol was found to be stereoselective in rat liver microsomes in vitro, preferentially metabolized (-)-enantiomer. The stereoselectivity could be influenced by the interaction between the two enantiomers and the enzyme selective inhibitors. PMID- 12579755 TI - [Inhibitory effect of melatonin on morphine withdrawal syndromes and the content of NO in plasma and brain tissue in morphine dependent mice]. AB - AIM: To observe the effect of melatonin (MT) on morphine withdrawal syndromes and determine the content of NO in plasma and brain tissue in morphine dependent mice. METHODS: A physical dependent model in mice was established by subcutaneous injection of morphine. MT (15 mg.kg-1, qd x 3) was given by intragastric infusion (ig) for three days. Withdrawal syndromes were induced by intraperitoneal injection of naloxon (5 mg.kg-1). The intensity of withdrawal syndromes was evaluated according to the jumping latency, the jumping times and the body weight loss. The content of NO was detected with Griess method. RESULTS: The jumping latency of morphine withdrawal reaction was prolonged and the jumping times were reduced obviously by ig MT. The increased NO content in plasma and brain tissue in morphine dependent mice was reduced by ig MT. CONCLUSION: The physical withdrawal syndromes and the content of NO in plasma and brain tissue in morphine dependent mice are inhibited by MT. PMID- 12579756 TI - [Effect of osthol on memory impairment of mice in AlCl3-induced acute senile model]. AB - AIM: To study the protective effect and mechanism of osthol on learning and memory impairment of mice with acute senile model induced by AlCl3. METHODS: After s.c. AlCl3 60 mg.kg-1 for 7 d and i.p. osthol 15 and 7.5 mg.kg-1 for 12 d, using step-through test and step-down test, the effect of osthol on learning and memory was observed and the glutathione peroxidase (GSH-PX) activities in blood and superoxide dismutase (SOD) activities in plasma and cerebrum were measured. RESULTS: Osthol 15 and 7.5 mg.kg-1 significantly improved the capability of memory and enhanced the activities of GSH-PX and SOD in AlCl3 treated mice. CONCLUSION: Osthol shows protective effect on brain memory impairment of mice in acute senile model induced by AlCl3. Perhaps the mechanism is involved in enhancing the activities of GSH-PX and SOD, clearing away the free radical, protecting the brain neuron from the harm of lipoperoxide. PMID- 12579757 TI - [Synthesis and anti alpha-adrenoceptor activity in vitro of 4-((4-((2 substitutedphenoxy)ethyl)-1-piperazinyl) methyl)- 1,2-dihydro-quinoline-2-one compounds]. AB - AIM: To study the synthesis and anti alpha-adrenoceptor activity of 1,2-dihydro quinoline-2-one compounds. METHODS: Acylation, bromination and cyclization, and substitute reactions were used in the synthesis of the title compounds IV. A proposed mechanism was showed to explain the unusual compounds 5 and 6 in the route C. The inhibition activity of the six target compounds were tested. RESULTS: Twelve new compounds were synthesized (II1-6 and IV1-6). Among them, six new compounds (IV1-6) are the title compounds. Structure of the title compounds were determined by IR, 1HNMR, MS and HRMS. CONCLUSION: Compounds IV3, IV4 and IV6 showed inhibitiion activity, and were worth further studying. PMID- 12579758 TI - [Two flavonoids from Lagopsis supina]. AB - AIM: To study the chemical constituents of Lagopsis supina. METHODS: Compounds were isolated by column chromatography of silica gel and Sephadex LH-20, and the structures were determined by spectral analysis. RESULTS: Two compounds were isolated and elucidated as apigenin-7-O-(6"-(E)-p-coumaroyl)-beta-D galactopyranoside (I) and apigenin-7-O-(3",6"-di-(E)-p-coumaroyl)-beta-D galactopyranoside (II). CONCLUSION: I and II are new compounds. PMID- 12579759 TI - [Chemical studies on SC3, a polysaccharide from Salvia chinensis]. AB - AIM: To study the chemical structure of SC3, an acidic polysaccharide from Salvia chinesis. METHODS: Based on chemical (including sugar composition analysis, methylation analysis, uronic acid reduction and partial acid hydrolysis) and spectral analysis (IR, NMR, ESI-MS), the structural characterization of SC3 was investigated. RESULTS: SC3 was composed Rha, Ara, Gal and GalA, with its mean molecular weight of 7.7 x 10(4). By means of methylation analysis, partial acid hydrolysis, NMR and ESI-MS spectrum, the linkages and sequence information of SC3 were obtained. CONCLUSION: SC3 is an complicated acidic polysaccharide, obtained for the first time from the plant. PMID- 12579760 TI - [A new bibenzyl derivative from Bletilla striata]. AB - AIM: To study the chemical constituents of Bletilla striata. METHODS: Various column chromatographies with silica gel and Sephadex LH-20 were employed for the isolation and purification. The structures of the compounds were elucidated on the basis of spectral analyses and chemical methods. RESULTS: Three compounds were isolated from the roots of Bletilla striata (Thunb.) Reichb. f. and identified as 5-hydroxy-4-(p-hydroxybenzyl)-3'-3-dimethoxybibenzyl (I), schizandrin (II), 4,4'-dimethoxy-(1,1'-biphenanthrene)-2,2',7,7'-tetrol (III). CONCLUSION: Compound I is a new bibenzyl derivative and II was isolated from this plant for the first time. PMID- 12579761 TI - [Studies on flavoniod constituents of Hedysarum multijugum]. AB - AIM: To study the chemical constituents from Hedysarum multijugum. METHODS: The compounds were separated by chromatography methods, their structures were identified by spectral analysis. RESULTS: Six compounds were isolated and identified as beta-sitosterol (1), 7-hydroxy-4'-methoxy isoflavone (2), betulic acid (3), 1,7-dihydroxy-3, 9-dimethoxy pterocarpene (4), 5, 7-dihydroxy-8-C prenyl-4'-methoxy isoflavone (5) and 5, 7-dihydroxy-4'-methoxy isoflavone (6). CONCLUSION: Compound 4 is a new compound and the others were obtained from the plant for the first time. PMID- 12579762 TI - [Studies on the chemical constituents of Coeloglossum viride (L.) Hartm. var. bracteatum (Willd.) Richter]. AB - AIM: To study the chemical constituents of the rhizomes of Coeloglossum viride (L.) Hartm. var. bracteatum (Willd.) Richter. METHODS: The compounds were mainly isolated with normal phase and reverse phase column chromatographic methods and HPLC. Their structures were elucidated on the basis of physical constants and spectral analysis (UV, IR, EI-MS, positive and negative FAB-MS, APCI-MS, 1HNMR, 13CNMR, DEPT, 1H-1H COSY, HMQC and HMBC). RESULTS: Eight compounds were obtained from the ethanolic extract of the rhizomes of this plant, dactylorhin B (I), loroglossin (II), dactylorhin A (III), militarine (IV), coelovirin A (V), gastrodin (VI), thymidine (VII) and quercetin-3,7-di-O-beta-D-glucopyranoside (VIII). CONCLUSION: All the compounds were obtained from this plant and genus Coeloglossum for the first time. V named coelovirin A, is a new compound. PMID- 12579763 TI - [Analysis of the response factors of different aminoglycoside antibiotics detected by evaporative light-scattering detector]. AB - AIM: To analyze if the response factors of different aminoglycoside antibiotics detected by evaporative light-scattering detector (ELSD) are the same. If they are, then ELSD can be applied to the quality analysis of this class of antibiotics. METHODS: The response factors of five different aminoglycosides (amikacin, sisomicin, netilmicin, etimicin and vertilmicin) detected by ELSD were determined by using a Diamonsil C18 column (150 mm x 4.6 mm, 5 microns) as analytical column and 0.2 mol.L-1 trifluoroacetic acid-methanol (94:6) as mobile phase at a flow rate of 0.6 mL.min-1, the temperature of the drift tube was set at 110 degrees C, and the flow of carrier gas at 2.80 L.min-1. Detector responses (A) and the amount of injection of each substance (m) were fitted to the logarithmic regression: logA = blogm + loga. RESULTS: The linear regression equation obtained were amikacin: Y = 1.46X + 5.07, gamma = 0.9997; sisomicin: Y = 1.51X + 5.03, gamma = 0.9997; netilmicin: Y = 1.52X + 4.88, gamma = 1.000; etimicin: Y = 1.46X + 4.85, gamma = 0.9999; vertilmicin: Y = 1.41X + 4.90, gamma = 0.9998. The differences between them were negligible. CONCLUSION: Different aminoglycosides can give the same responses with ELSD detection. So, the HPLC ELSD methods can be applied to the analysis of impurities, the control of the ratio of multi-components drug and the determination of new substances by using another substance as reference, etc. PMID- 12579764 TI - [Determination of irisquinone by single sweep oscillopolarography]. AB - AIM: To propose a polarographic method for the determination of irisquinone. METHODS: A reduction wave of irisquinone was recorded by single sweep oscillopolarography. RESULTS: In 8.0 x 10(-3) mol.L-1 Na2B4O7-1.6 x 10(-2) mol.L 1 KH2PO4 (pH 7.7) supporting electrolyte, a redution wave of irisquinone with peak potential -1.23 V (vs SCE) achieved high sensitivity. The 2nd-order derivative peak current of the reduction wave was proportional to irisquinone concentration in the range of 1.5 x 10(-7)-5.2 x 10(-6) mol.L-1 (gamma = 0.9992, n = 9). The detection limit was 6.0 x 10(-8) mol.L-1. Relative standard deviation (RSD) was 0.87% by performing 13 independent measurements on 2.0 x 10(-6) mol.L-1 irisquinone. CONCLUSION: The proposed method was sensitive, simple, rapid, and can be applied to the determination of irisquinone in raw medicine and capsule. PMID- 12579765 TI - [Pharmacokinetics and relative bioavailability of probucol inclusion complex capsule in healthy dogs]. AB - AIM: To study the pharmacokinetics and relative bioavailability of probucol inclusion complex capsule. METHODS: Following oral administration of a single dose of 250 mg of conventional tablet (formulation A, purchased from the market) and probucol inclusion complex capsule (formulation B, a new formulation for preclinical trial) to each of 6 healthy dogs in a randomized crossover design, the plasma levels of the active drug at different time points were determined by HPLC and the plasma concentration-time profiles of formulation A and B were obtained. The pharmacokinetic parameters as well as relative bioavailability were analyzed. RESULTS: The concentration-time curves of formulation A and formulation B were found to fit a two-compartment open model. The Tmax values of formulation A and formulation B were (9.3 +/- 2.1) h and (9.3 +/- 2.1) h, the Cmax values were (1.5 +/- 1.0) microgram.mL-1 and (2.3 +/- 0.9) microgram.mL-1 and the AUC0 240 values were (85 +/- 56) microgram.h.mL-1 and (134 +/- 55) microgram.h.mL-1, respectively. The relative bioavailability of formulation B was found to be (198 +/- 90)% compared with formulation A. The results of variance analysis and two one-side t-test showed that there was significant difference between the two formulations in the AUC0-240. CONCLUSION: The high bioavailability by the inclusion of formulation B is attributed to the improvement of its water solubility by the inclusion process and this is supposed to be a key factor for improving drug bioavailability. PMID- 12579766 TI - [Capillary zone electrophoresis for determination of thymulin in human plasma]. AB - AIM: To establish a capillary zone electrophoresis (CZE) method for determination of thymulin in human blood plasma. Thymulin is a nonapeptide produced by the thymic epithelium. METHODS: The uncoated capillary used was 75 microns ID x 57 cm (50 cm effective length). The experimental conditions were as follows: the running buffer was phosphate-borate buffer containing 50 mmol.L-1 Na2B4O7-10 mmol.L-1 Na2HPO4 (pH 9.10); running voltage was 12 kV; operated temperature was 25 degrees C; running time was 13 minutes, detective wavelength was 200 nm. RESULTS: The linear range of the thymulin standard curve was 1-10 micrograms.mL 1, gamma = 0.9990, the average recovery was 70.97% with RSD of 3.32%, the RSD of reproducibility was 3.05%. CONCLUSION: The method of CZE for determination of thymulin in human plasma is simple and reliable. PMID- 12579767 TI - [Separation of bile acids by capillary zone electrophoresis]. AB - AIM: To develop a method for separating the major bile acids by capillary zone electrophoresis (CZE). METHODS: The effect of different separations, such as the compose, pH and the concentration of buffer, on the electro-osmotic flow (EOF), the migration time and resolution of 8 bile acids in this system were studied. The general trends in migration time could be correlated to the pH and concentration of the buffer. The effect of organic reagent on EOF and migration time were also investigated. By addition of methanol, the EOF went smaller than before, and better resolution was achieved. The experimental results showed that optimum separation was achieved under the following condition: buffer composition of 126 mmol.L-1 disodium tetraborate, 43 mmol.L-1 disodium hydrogenphosphate, 18% methanol; temperature 30 degrees C; voltage 30 kV; total length of capillary 570 mm and 500 mm from injection end; ultraviolet detection at 200 nm; pressure injection 5 kPa for 8 s. RESULTS: Eight kinds of bile acid had been separated by CZE with only one injection. The method was used to analyse the contents of bile acids from different kinds of bear biles, the recovery was 89%-107%. CONCLUSION: This method is simple and rapid, and can be used to determine the content of bile acids in bear biles. The calibration curve showed good linearity for eight bile acids in the concentration range of 4-60 mg.mL-1 (gamma > 0.9954). The total time for seperation and determination was within 25 min. PMID- 12579768 TI - [Studies on pulsatile release tablets of diltiazem hydrochloride in explosion way]. AB - AIM: To investigate the preparation of pulsatile release tablets, the release of the drug in vitro and the pharmacokinetics in vivo. METHODS: Diltiazem hydrochloride (DIL) was used as model drug. The pulsatile release tablets were prepared by film-coated method using ethylcellulose and Eudragit L. The effect of formulation on pulsatile release of diltiazem hydrochloride was investigated under release rate test. The mechanism of pulsatile release of drug was proved by the test of water-uptake. The pharmacokinetic and bioavailability study in eight human subjects was performed by HPLC method. RESULTS: The release of diltiazem hydrochloride effected by the formulation of the core tablets and the composition and thickness of the coating film. In vitro, the delayed-release time T10 was 4.4 h, the maximum release time Trm was 8.0 h and the pulsed-release time Trm-10 was 3.6 h. In vivo, the delayed-release time Tlag was 4.9 h, the peak time was 8.0 h and the pulsed-release time was 3.1 h. The relative bioavailability was 105%. CONCLUSION: The release of drug from pulsatile release tablets of diltiazem hydrochloride was shown to be in pulsed way both in vitro and in vivo. PMID- 12579769 TI - [In vitro drug release profiles and mucoadhesive property of bioadhesive microspheres of metronidazole]. AB - AIM: To prepare bioadhesive microspheres of metronidazole (Metro) with prolonging resident time in the stomach and sustaining drug release. METHODS: The microspheres were prepared by a drying-in-liquid method. The appearance, particle size and drug release in vitro were examined. The factors influencing bioadhesive property and drug release, such as ethyl cellulose (EC)/carbopol 934P (CP) ratio, particle size and Metro content were investigated. RESULTS: The average diameter of the Metro-EC-CP microspheres was 559.9 microns. The release profiles of metronidazole were shown to fit to first-order equations well. With the increase of CP content in the Metro-EC-CP microspheres, the microspheres showed better mucoadhesion and faster drug release. The drug release rate decreased with the increase of particle size and the decrease of Metro content. CONCLUSION: The Metro-EC-CP microspheres have a sound mucoadhsive property and sustained drug release when the ratio of EC and CP was 17:3 and Metro content was 25%. The drug release was shown to last for 8 h in 0.1 mol.L-1 hydrochloric acid. PMID- 12579770 TI - [Tyrosine kinase receptor-mediated signal transduction and cancer treatment]. PMID- 12579771 TI - [Progress in studies on SOD mimics]. PMID- 12579772 TI - [The effect of transforming growth factor-beta 1 on the secretion of collagen type IV in cultured human trabecular cells]. AB - PURPOSE: To investigate the effect of transforming growth factor-beta 1 (TGF-beta 1) on the expression of collagen type IV in cultured human trabecular cells. METHODS: On the basis of successful culture of human trabecular cells, we detected the content of collagen type IV in the media and covers of trabecular cells, treated with TGF-beta 1 (1 ng/ml), by using enzyme-linked immunosorbent assay(ELISA) and immunohistochemical techniques respectively. RESULTS: ELISA method demonstrated that, in the first four days, there was statiscally significant difference between TGF-beta 1 treated and control groups with the collagen type IV content of 187.50 +/- 29.01 pg/ml and 93.75 +/- 20.50 pg/ml respectively, then in the second four days, there was no statistically significant difference between experimental group of 128.75 +/- 52.02 pg/ml and control group of 91.25 +/- 21.36 pg/ml. Neither statistical difference was found between two treated groups, nor was between two control groups. Immunohistochemical techniques showed, the positive staining of collagen type IV was stronger in cell covers of experimental group treated with TGF-beta 1 for 12 days than that of control group. CONCLUSION: TGF-beta 1 can increase the expression of collagen type IV in cultured trabecular cells, it may be among causes of abnormal accumulation of extracellular matrices in trabecular meshwork of eyes with primary open angle glaucoma. PMID- 12579773 TI - [The cultivation, freezing-state preservation and resuscitation of human iris pigment epithelial cells in vitro]. AB - PURPOSE: To establish the culture of human iris pigment epithelial cells in vitro and study the freezing-state preservation and resusciation. METHODS: The iris pigment epithelial specimens from human eyes were cultured in vitro. Based on the cell-frozen principle, the cultured cells were collected, then frozen in liquid nitrogen and resuscitation. RESULTS: Cultured iris pigment epithelial cells were obtained. Under the inverted light microscope, primary cells appeared multigonal and arranged in monolayer, there were abundant pigment granules in the cytoplasm and the nuclei each of which contained 1 or 2 nuceoli wre relatively transparent. Under the transmission electron microscope, there were plenty of microvilli at the cell membrane and desmosomes present in the intercellular space. 6 groups of cells were frozen. The resuscitation exeriment was carried out for 4 times, every times being successful. All of the resuscitated rates were more than 90%. CONCLUSION: The human iris pigment epithelial cells were cultured in vitro successfully, they can also be frozen and resuscitated, which will be useful in studies of pathogenesis for some eye disease. PMID- 12579774 TI - [The histological changes after preserved human amniotic membrane transplantation for conjunctival reconstruction of rabbits eyes]. AB - PURPOSE: To investigate the histological changes after preserved human amnotic membrane transplantation of rabbit eyes. METHODS: The conjunctiva of 12 New Zealand albino rabbits' right eyes were excised partly. Then the preserved human amniotic membrane were transplanted on the sclera. Eyes were enucleated at 1 week, 3 weeks, 5 weeks and 12 weeks (n = 3 for each) after surgery, and they were examined histologically with staining of HE, PAS, and transmission electronic microscopy. RESULTS: At 1 week after transplantation, the human amniotic membrane firmly attached on the sclera, with the dead epithelial cells remaining on its surface. At 3 weeks, conjunctival epithelization was completed on the human amniotic membrane. Abundant PAS-positive goblet cells were recognized. The amnion began resolve. At 12 weeks, the amnion dissolved. On transmission electronic microscopy, the cell polarity is prominent. The epithelial cells contained a large number of mitochondria and endoplasmic reticulum. Microvilli were pronounced on the apical cell membrane. There are many intracellular desmosomes and hemidesmosomes. CONCLUSION: Preserved human amniotic membrane is a valuable conjunctival replacement material. After transplantation, the histology of new tissue is as same as normal conjunctive. PMID- 12579775 TI - [Effects of epidermal growth factor on wound healing of penetrating keratoplasty in rabbits]. AB - OBJECTIVE: To observe the effects of epidermal growth factor (EGF) on wound healing of the penetrating keratoplasty(PKP) in rabbits. METHODS: Twelve rabbits (24 eyes) were stratified randomly divided into 6 groups and underwent penetrating keratic autoplasy. Liquid scintillation counter was used to measure the incorporation rate of 3H-TdR. Pressure-detector was used to survey the intensity of keratic wound healing. AgNORs staining was used to count the fibroblast cells at keratic wound. HE staining, VG staining, AgNORs staining and transmission electron microscopy were used to observe the morphological changes of keratic wound. RESULTS: 1 The intensity of EGF group on 8 days, 14 days and 21 days after PKP was significantly higher than that of the control group(P < 0.01). 2. The incorporation rate of 3H-TdR in wound healing the EGF group on 14 days and 21 days was obviously higher than that of the control group(P = 0.01). 3. The number of fibroblast cells of EGF were more than that of the control group(P < 0.01) in 8 days. CONCLUSION: EGF can promote the wound healing in rabbits after PKP. PMID- 12579776 TI - [Protective effects of brain-derived neurotrophic factor on injured retinal ganglion cells of rats]. AB - PURPOSE: To investigate in vivo the survival of retinal ganglion cells(RGC) after optic nerve crush with intraocular injection of Brain-derived neurotrophic factor (BDNF). METHODS: Sprague-Dawly(SD) rats were divided into normal control, crush control, medium treatment and BDNF treatment group, and every group had 20 eyes. RGC of adult rats were labeled with fluorogold. Seven days later, the optic nerve was intraorbitally crushed and BDNF was injected into the vitreous. Five, 7, 14, 21 and 28 days after crush, the numbers of RGC were counted. RESULTS: The densities of RGC began to decrease 7 days after injury, the number of RGC was 70.2% and 40.5% of normal controls on days 14 and 28, respectively. In the group with BDNF injection, RGC densities decreased on days 7, but RGC densities were much higher than that of controls on 7, 14, 21, and 28 days after injury (P < 0.01). CONCLUSION: BDNF administered intraocularly at the time of crush can protect RGC from injury and cell death. PMID- 12579777 TI - [mRNA expression of keratoepithelin in cornea after photorefractive keratectomy in rabbits]. AB - OBJECTIVE: To investigate the roles of keratoepithelin(KE) during healing after photorefractive keratectomy (PRK). METHODS: 20 of 24 rabbits underwent bilateral 193 nm excimer laser photorefractive keratectomy to correct 10 diopters of myopia, and the other 4 rabbits as normal group. The mRNA levels of KE in corneas of the rabbits were determined by the in situ hybridization technique with a Digoxigenin labled probe. RESULTS: In normal group, there was no mRNA expression of KE in cornea. on 7, 14 days after operation, mRNA expression of KE was observed in epithelium and stroma of operation group, while on 28 days mRNA expression of KE was observed only in epithelium. CONCLUSION: KE was involved in the synthesis of extracellular matrix during healing after PRK and may play an important role in the development of haze by maintaining proper spacing among collagen fibrils. PMID- 12579778 TI - [Effect of amniotic membrane on expressions of TGF-beta 1, collagens I, III and fibronectin in rabbit corneal healing after photorefractive keratectomy]. AB - OBJECTIVE: To observed the effect of amniotic membrane transplantation (AMT) on expressions of transforming growth factor-beta 1(TGF-beta 1), collagens I, III and fibronectin (FN) in rabbit corneal healing after photorefractive keratectomy (PRK), and investigate the anti-scarring formation mechanism of AMT. METHODS: Ten rabbits underwent bilateral PRK to correct 8 diopters of myopia. One eye was randomly transplanted with preserved human amniotic membrane, and the other eye served as the control. The expressions of TGF-beta 1, collagens I, III and FN were studied by immunohistochemistry at 4 weeks after operation. RESULTS: The expression of TGF-beta 1 in corneal epithelium and keratocytes and collegans in anterior stroma was significantly less in AMT group than in the control group. The expressions of collagens III and FN in anterior stroma of ablation area were also significantly less in AMT group than in the control group. The staining of collagen I had no significant difference between the AMT group and the control group. CONCLUSION: The expressions of TGF-beta 1, collagens III and FN can be suppressed by amniotic membrane after PRK. These data suggest that part of the anti-scarring effect of AMT may be mediated through the suppression of TGF-beta 1, collagens III and FN expression. PMID- 12579779 TI - [The muscarinic receptor subtype study of the action on lowering IOP of cholinomimetic drugs]. AB - PURPOSE: Clarify the relationship between lowering intraocular pressure (IOP) action of cholinomimetic drugs and muscarinic receptor subtypes. METHODS: Measuring rabbit IOP with gas driving ophthalmotonometer, measuring size of pupil with pupil rule and measuring contraction of rabbit iris by iris experiment in vitro. Figuring out pD2 of cholinomimetic drugs and pA2 of the antagonists of muscarinic receptor subtypes. RESULTS: The order of effect on lowering IOP and miosis of cholinomimetic drugs is Erycibele Alkaloid > Aceclidine > Pilocarpine and the order of effect on constracting iris is Erycibele Alkaloid > Pilocarpine > Aceclidine. The sequence of antagonistic effect of subtype antagonists is 4 DAMP(M3) > Pirenzepine(M1) > Gallamine(M2). CONCLUSION: Effect cholinomimetics on lowing IOP and miosis is primary mediated by M3 receptor, next by M1 receptor and has little relation with M2 receptor. PMID- 12579780 TI - [The study on superoxide dismutase and trace element in patients with senile cataract]. AB - PURPOSE: To investigate the relation of senile cataract with superoxide dismutase(SOD) and trace elements. METHODS: Superoxide dismutase of red blood cells and trace elements(Cu, Zn, Ca, Fe) of serum in 61 patients with senile cataract and 121 normal persons were investigated. RESULTS: The activity of superoxide dismutase in patients with senile cataract decrease dramatically in comparison with normal control, and there was no statistical difference about trace elements of Cu, Zn, Ca between senile cataract and control group. But Fe element in patients is higher than that of the normal group. CONCLUSION: The results support the hypothesis that the formation of senile cataract is capable of antioxidative scavenger system of lens decreases with age. PMID- 12579781 TI - [Simultaneously corneal laceration repair, traumatic cataract removal and intraocular lens implantation]. AB - OBJECTIVE: The study aimed to evaluate the outcome of patients who undrwent simultaneously corneal laceration repair, cataract extraction and posterior chamber lens implantation. METHODS: Traumatic cataract extraction and posterior chamber intraocular lens (PC IOL) implantation were performed in 36 eyes of 36 cases with corneal laceration. YAG laser was the postoperative procedure for embrane before the lens and after cataract. Penetrating keratoplasty was performed for central cicatrice of cornea. All the patients have been followed up for 3-12 months. RESULTS: The postoperative visual acuity s.c. was 0.1-0.3 in 8 cases(22.8%), 0.4-0.5 in 14 cases (40%) 0.6-0.8 in 13 cases (37.1%) on following up. There were no serious complication attributable to PC IOL implantation. CONCLUSION: As long as the range of surgical indication was known well, PC IOL implantation can get good clinical effects at the time of primary repair. PMID- 12579782 TI - [Phacoemulsification in white cataract]. AB - OBJECTIVE: To evaluate the value and technique of Phacoemulsification in white cataract. METHODS: Ninty eyes of senile cataract were divided into two groups by the density of the nuclei. Different operative technique was done in two groups. RESULTS: The success rate of the continuous curvilinear capsulorhexis was 88.9%. Seventy five eyes have got the visual acuity over 0.5 on the first day after the operation. There was significant difference between two groups. The complications of operation were mild corneal edema and posterior capsule tear. CONCLUSIONS: It's difficult to complete the phacoemulsification of white cataract. The surgeons are required the skillful technique and experience. However, phacoemulsification is valuable in white cataract. PMID- 12579783 TI - [The influence of experimental scotoma on multifocal electroretinogram]. AB - PURPOSE: To detect the sensitivity and the repeatability of multifocal electroretinogram (mfERG). METHOD: The effects of experimental absolute and relative scotomas on mfERG with 103 hexagons were observed on 8 normal eyes. The scotomas were produced by simultaneous and multi located masks on the hexagons of the screen in 6 different locations with black paper, and the neutral filters of 0.3 and 0.6 log unit. The repeatability was evaluated by analysis the data from 5 unmasked hexagons in 4 successive tests. RESULTS: When the hexagons were 1/2 or wholly masked with the black paper and the two kinds of the filters, the amplitudes of N1 and P1 waves in these areas were significantly decreased as with the reduced transferability of light. When the hexagon was 1/3 masked, there were no significant differences between the different kinds of mask and the unmasked control. There were no significant changes in the datas from the selected unmasked hexagons in the 4 successive recordings. CONCLUSION: mfERG is an unique sensitive visual electrophysiological test with high sensitivity and good repeatability. When 103 hexagons stimulating pattern is used, it could detect the scotoma larger than 1/3 of each hexagon. PMID- 12579784 TI - [The Ahmed glaucoma valve implantation for refractory glaucoma]. AB - OBJECTIVE: To evaluate the efficacy of the Ahmed Glaucoma Valve implant in the treatment of refractory glaucoma. METHODS: 28 cases(29 eyes) of refractory glacucoma were treated by the Ahmed glaucoma Valve implant, including 10 cases (10 eyes) of neovascula glaucoma, 5 cases (6 eyes) of juvenile glaucoma and 13 cases (13 eyes) of other causes glaucoma. RESULTS: Post-operative follow-up time was 3-12 months. The intraocular pressure (IOP) was lowered from preoperative 40.9 +/- 7.32 mmHg (1 mmHg = 0.133 kPa) to postoperative 20.63 +/- 2.75 mmHg in Neovascular glaucoma; from preoperative 41 +/- 5.66 to postoperative 18.00 +/- 2.12 mmHg in glaucoma after IOL implant combined with vitrectomy operation; from preoperative 33.65 +/- 5.28 to postoperative 16.45 +/- 1.48 mmHg in juvenile glaucoma; from preoperative 31.43 +/- 4.13 to postoperative 18.13 +/- 1.25 mmHg in the failure conventional filtering surgery of glaucoma. The postoperative complicatons were flat anterior chamber, hyphema, anterior iritis and so on. CONCLUSION: The Ahmed glaucoma valve implant is a useful method in lowing IOP of refractory glaucoma. PMID- 12579785 TI - [Clinic analysis of retinopathy in gestosis patients]. AB - PURPOSE: To explore the relationship between degree of retinopathy and degree of gestosis, hypertension, and fetal death rate by examing eye fundus of 115 gestosis patients. METHOD: We compared the degree of retinopathy with the degree of gestosis, the degree of hypertension, and the fetal death rate respectively. RESULTS: The degree of retinopathy is postively related with the degree of gestosis, the degree of hypertension and the fetal death rate. CONCLUSION: Eye fundus examination in gestosis patients is an significant guideline for clinical treatment. PMID- 12579786 TI - [Application of cellulose acetate membrane with the technique of impression cytology combined with immunohistochemical staining for detection of dry eye]. AB - PURPOSE: To evaluate the conjunctival cells by impression cytology with cellulose acetate membrane. METHODS: The expression of the TGF-beta 1 in conjunctival epithelium of 24 patients with dry eye were studied by impression cytology combined with immunohistochemical staining and observed by microscope. RESULT: The cellulose acetate membrane is very transparent under the microscope. Cell's membrane by TGF-beta 1 staining is clear, cytoplasm brown and nuclei blue. CONCLUSION: The method of impression cytology with cellulose acetate membrane is simple and has no injury to the ocular surface. Combined with immunohistochemistry staining, it is suitable to observe the conjunctival cells by this technique which has an important value for the examination of the ocular surface disease. PMID- 12579787 TI - [Clinical analysis of evisceration with hydroxyapatite implant]. AB - PURPOSE: To evaluate the clinical effect and safety of evisceration with hydroxyapatite (HA) orbital implant. METHODS: A retrospective analysis of all eviscerations with HA implant performed between January 1998 and May 2000 in Zhongshan Ophthalmic Center was completed. Patient records were reviewed for demographic data, surgical indication, sphere size, clinical outcome, and complications. The average follow-up interval was 11.6 months (2-24 months). RESULTS: Primary evisceration with HA implantation was performed in total 78 cases. All patients underwent successful surgery without major complications. There was no case with those complications, including implant extrusion, exposure and implant dislocation, in the series examination. No patient has symphathetic ophthalmia during follow-up interval. CONCLUSION: In strict selected cases with surgical indication of evisceration, primary evisceration with HA implantation is a safe and effective method for treating patients with a blind, painful or atrophic eye. PMID- 12579788 TI - [Ocular axial length and refractive changes in pediatric pseudophakia]. AB - PURPOSE: To evaluate the ocular axial length and refractive changes that occured in children who had cataract extraction with intraocular lens implantation and to investigate the factors that influenced the pseudophakic refraction. METHODS: A review of 12 eyes in 10 children (mean age 7.16 years, range 3.83 to 10.16 years) who had cataract extraction with intraocular lens implantation was undertaken. Patients were followed for an average of 35.8 months. The preoperative and the last postoperative axial length and corneal curvature (K readings) in both operated and unoperated eyes were measured. The distance from the vertex of the cornea to the anterior vertex of intraocular lens (AVpc) was measured in the final follw-up. The initial (in two weeks) and the last refractive status of pseudophakia were examined. Overall age at surgery averaged 7.16 years (range 3.83 to 10.16 years), with followed up of 35.8 months. RESULTS: In the 12 operated eyes, the mean axial growth was 0.39 mm, whereas in the other eight unoperated eyes it was 0.66 mm. Though the axial length of both eyes increased significantly after surgery (P < 0.05), there was no significant difference in the postoperative increase of axial length between the two groups(P > 0.05). In K readings, there was no significant difference between the operated and unoperated eyes before surgery and in the last follow-up respectively. There was no significant difference between the preoperation and the last follow-up in the operated and the unoperated eyes respectively. The average calculated AVpc (AVpc2) was 4.66 30 mm, but the average objective AVpc (AVpc1) measured with ultrasonic biometry was 3.83 mm. The average difference between them was 0.8320 mm and was significant (P < 0.01). According to SRK formula, using preoperative and the last postoperative axial length and corneal curvature to predict refractive changes, the myopic shift was -1.53 D, but the average objective refractive difference between the last and initial examination after surgery was 3.86 D. The average difference between the predicted and actual postoperative refractive changes was significant (P < 0.05). CONCLUSION: Our study suggests that there may be no effect on ocular growth in children (3 to 10 years) followed cataract extraction and intraocular lens implantation. Increasing of axial length and moving forward of intraocular lens induced by complication may result in myopic shift in pseudophakia of children. Slightly undercorrected eyes with intraocular lens in children after cataract extraction will gradually move to emmetropia or moderate myopia in adulthood. PMID- 12579789 TI - [Multiple factors analysis of myopic regression after photorefractive keratectomy]. AB - PURPOSE: To investigate myopic regression after excimer laser photorefractive keratectomy(PRK) in patients with myopia. METHODS: PRK was performed to correct myopia errors from 1.00 D to 16.00 D diopters on 214 myopia eyes(121 patients). All eyes were divided into two groups: group I--1.00 D(-)-6.00 D (124 eyes), group II--6.25 D(-)-16.00 D(90 eyes). All the patients were followed up for two years. RESULTS: A Significant positive correlation was found between the regression and the extents of attempted correction. The rate of regression in mild to moderate myopia group was 9.7% while in highter group it was 27.8% (P < 0.005). The rate of regression of older age group in lighter myopia group was highter than that in younger case (P < 0.005). Relatively severe haze under the corneal epithelium often accompanies the regression. CONCLUSION: The main factors which caused the myopia regression after PRK was the extent of attempted correction, patients' age and corneal haze. PMID- 12579790 TI - [The analysis of the axises of corneal astigmatism and total astigmatism in astigmatic patients]. AB - PURPOSE: The axial amount and the relationship between the axises of corneal astigmatism and total astigmatism were statistically analyzed in ametropic patients, for the purpose of selecting the refractive correction, the base curve of contact lens, the axis of astigmatic contact lens, and providing the reference of operational amount of ecimer laser keratoplasty. METHODS: The refractive state and the corneal curvature of ametropic patients were measured and consecutvely repeated 3 times in the same eye at the same state with the same autorefractometer in 107 cases. The detected axises of total astigmatism and corneal astigmatism were statistically analyzed through the computerized recording. RESULTS: The mean angle between the axis of corneal astigmatism and the axis of total astigmatism was 5.73 degrees +/- 3.97 degrees in myopic patients, 3.13 degrees +/- 2.49 degrees in mixture astigmatism patients and 6.35 degrees +/- 4.20 degrees in hyperopic patients. CONCLUSION: The angle between the axis of corneal astigmatism and the axis of total astigmatism in the refractive state was mainly caused by the difference of the axises of surface astigmatism of lens and cornea. PMID- 12579791 TI - [Effect of praeruptorum caumarin on cardiac mass, myocardial [Ca2+]i and Na+, K(+)-ATPase, Ca2+, Mg(2+)-ATPase activity in renovascular hypertensive rats]. AB - AIM: To investigate the preventive and reversional effect of praeruptorum caumarin compound on left ventricular hypertrophy in renovascular hypertensive rats (RHR) and its mechanism. METHODS: The two-kidney-one-clip (2K1C) RHR model was used. The blood pressure, wet weight of the left ventricle, surface area of myocardial cells, resting [Ca2+]i level and Na+, K(+)-ATPase, Ca2+, Mg(2+)-ATPase activity of myocardial membrane and mitochondria were measured. RESULTS: Praeruptorum caumarin 30 mg.kg-1.d-1 was given ig for 9 weeks from the 6th or 9th week after operation in the preventive or regressive group. The blood pressure, left ventricle wet weight and area of myocardial cells of the preventive and regressive group were significantly reduced than that of the LVH group. The resting [Ca2+]i of the both praeruptorum caumarin treated groups (121 +/- 13, 133 +/- 9 nmol.L-1) were lower than that of the LVH group (158 +/- 7 nmol.L-1). The KCl-induced [Ca2+]i elevation was decreased more significantly in preventive and regressive group than that of the hypertrophic myocytes. The activity of Na+, K(+)-ATPase and Ca2+, Mg(2+)-ATPase increased by 40% and 93% in the preventive group, 28.4% and 48.8% in regressive group than that of the LVH group. CONCLUSION: Praeruptorum caumarin was shown to prevent and reverse hypertrophy of LVH by lowering [Ca2+]i and increasing the ATPase activity. PMID- 12579792 TI - [Synergistic antitumor effect of sodium caffeate and mitomycin]. AB - AIM: To study the synergistic antitumor effect of sodium caffeate (sodium 3,4 dihydroxycinnamate, CA-Na) and mitomycin C (MMC). METHODS: MTT assay, Western Blotting analysis and flow cytometry were used to determine the effects of MMC alone and in combination with CA-Na on tumor cell proliferation, cell cycle progression and the expression of apoptosis-involved proteins Bcl-2 and caspase 3. Intracellular Ca2+ level and mitochondria membrane potential were also assayed. Inhibition of tumor growth was evaluated with hepatoma 22 transplanted s.c. in mice. RESULTS: Synergistic inhibition of hepatoma BEL-7402 cell proliferation was achieved by the combination of MMC and CA-Na. The combination of MMC and CA-Na caused more remarkable cell cycle perturbation in BEL-7402 cells than MMC alone. Combination of CA-Na and MMC synergistically inhibited Bcl-2 expression and enhanced caspase-3 expression of the cells. Intracellular Ca2+ level was significantly increased when the cells were treated with MMC combined with CA-Na. Mitochondria membrane potential was markedly decreased when treated with the combination. At doses of 0.5, 1.0 and 1.5 mg.kg-1, i.p., x 10, MMC inhibited the growth of hepatoma 22 by 35.9%, 62.7% and 79.1%, respectively. In combination with CA-Na, the inhibition rates of MMC increased to 62.4%, 70.5% and 88.0%, respectively. CONCLUSION: The combination of CA-Na and MMC showed synergistic effect against tumor cell proliferation in vitro and tumor growth in vivo. PMID- 12579793 TI - [Effects of benazepril on apoptosis in the kidney of diabetic rats]. AB - AIM: To investigate the effects of angiotensin converting enzyme inhibitor (ACEI) on apoptosis and the expression of Fas and Fas-L in the kidney of diabetic rats. METHODS: Uninephrectomized Spraque-Dawley rats were used to induce diabetes by intraperitoneal injection of streptozotocin (65 mg.kg-1). Benazepril (10 mg.kg-1) was given daily by gavage from the next day of the induction to diabetes for 12 weeks. Apoptosis was evaluated by means of terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL). Flow cytometry and immunohistochemistry were used to detect the expression of Fas and Fas-L. RESULTS: Compared with those in the kidneys of control group, apoptotic cells were more in number and the expression of Fas and Fas-L was higher in the diabetic kidneys (P < 0.05). The number of apoptotic cells and the level of expression of Fas and Fas-L were reduced by benazepril treatment (P < 0.05). CONCLUSION: Angiotensin converting enzyme inhibitor benazepril showed some renal protective effect on diabetic nephropathy, partly through inhibiting apoptosis by down-regulating Fas and Fas-L expression. PMID- 12579794 TI - [The protective effect of bicyclol on ischemia-reperfusion induced kidney injury in rats]. AB - AIM: To investigate the protective effect of bicyclol on kidney injury induced by ischemia-reperfusion in rats. METHODS: Bicyclol was orally administered to rats at doses of 50 and 200 mg.kg-1 before ischemia-reperfusion injury to evaluate the influence of bicyclol on the formation of MDA and BUN in serum, the content of GSH and the activity of GST in kidney, as well as kidney mitochondria membrane fluidity in ischaemia-reperfusion rats. RESULTS: Bicyclol given orally at doses of 50 and 200 mg.kg-1 was shown to significantly decrease the increment of MDA and BUN in serum and protect the GSH depletion in kidney. Bicyclol was also shown to induce kidney GST and ameliorate the decrease of mitochondria membrane fluidity. The protective effects of bicyclol on kidney injury induced by ischemia reperfusion are dose-dependent. CONCLUSION: The protective action of bicyclol on kidney injury induced by ischemia-reperfusion may be attributed to its induction of kidney GSH and the GST, stabilization on mitochondria membrane and the inhibition on lipid peroxidation. PMID- 12579795 TI - [Effects of donepezil on the delayed rectifier-like potassium current in pyramidal neurons of rat hippocampus and neocortex]. AB - AIM: To investigat the effects of donepezil on delayed rectifier-like potassium currents (IK) in rat hippocampus and neocortex. METHODS: Whole cell configuration of the patch-clamp techniques were used to characterize IK in acutely isolated rat hippocampal and neocortical pyramidal neurons. RESULTS: The slowly inactivating outward currents (IK) were recorded in all cells under investigation. Donepezil in micromolar concentrations were shown to supress the IK of all cells in a dose-dependent and voltage-dependent manner. The steady state activation curves of IK were characterized by half-activation potentials of -15.5 mV in hippocampal and -4.1 mV in neocortical pyramidal neurons and were changed to -26.2 mV and -18.6 mV, respectively, after perfusion with donepezil (10 mumol.L-1). CONCLUSION: At concentrations as low as 1 mumol.L-1, donepezil was found to block the IK in a voltage-dependent manner in hippocampus and neocortex. This effect may be synergistic with the anticholinesterase activity of donepezil to increase its therapeutic effectiveness. PMID- 12579796 TI - [Potentiation of radiosensitivity by staurosporine associated with abrogation of G2 phase arrest]. AB - AIM: To investigate the radiosensitizing effect and mechanism of action of staurosporine (STP) in human colon carcinoma HT-29 and breast cancer MCF-7/ADR cells. METHODS: The effect of STP on the cytotoxicity of X-ray was determined by clonogenic assay. The effect of STP on cell cycle arrest induced by X irradiation was studied in two cell lines by using flow cytometry, Western Blotting was performed to indicate the changes of cyclin B1 and cdc2 protein levels. RESULTS: STP sensitized the two cell lines to X-ray by clonogenic assay. STP potentiated the cytotoxicity of X-ray by 2.10- and 2.09-fold in HT-29 and MCF-7/ADR cells. Flow cytometry assay showed that exposure of HT-29 and MCF-7/ADR cells to X-ray caused cells arrest in G2 phase. The percentage of arrest G2 phase cells were 56% and 52.7%, respectively. The addition of STP after irradiation resulted in a dose dependent reduction of G2 phase arrest induced by X-ray. Furthermore, the results showed that STP blocked decrease of cyclin B1 expression induced by X-ray, while mitotic index measurement indicated that X-ray-irradiated cells treated with STP entered mitosis. The data suggested that the potentiation of cytotoxicity of X ray by STP is associated with the suppression of cyclin B1 expression, which result in the abrogation of G2 arrest, before the cells entered into M phase, they had not enough time to repair. CONCLUSION: STP is a potent G2 checkpoint abrogator and markedly enhanced the cytotoxicity of X irradiation in the p53 mutant cancer cells. PMID- 12579797 TI - [Pharmacokinetic study of recombinant human acidic fibroblast growth factor in rabbits by skin external use]. AB - AIM: To investigate the pharmacokinetic characteristics of recombinant human acidic fibroblast growth factor (rhaFGF) after external use in rabbits. METHODS: 125I-rhaFGF 180 U.cm-2 was daubed to normal skin and scathed skin in rabbits. The radioactivity and paper chromatography were used to determine the 125I concentrations and distribution in plasma and organs at different times. RESULTS: The plasma concentration of 125I-rhaFGF increased rapidily, and reach peak plasma level (73.03 pg.mL-1) thirty minutes after administration. Then the concentration of 125I-rhaFGF decreased quickly after thirty minutes, and approached to zero after three hours. Highest radioactivity accumulated in the skin, followed by kidney, lowest in the brain 96 h after administration. CONCLUSION: rhaFGF can not be absorbed from the normal skin, whereas a small amount of rhaFGF can be absorbed through scathed skin. The t1/2 of rhaFGF in plasma was very short. Cumulative effect of rhaFGF was not observed. Absorbed rhaFGF showed high affinity to skin, and can be distributed to skin far from the site of administration. PMID- 12579798 TI - [Synthesis of new N3S pseudo-peptide complexes and biodistribution in mice]. AB - AIM: To explore the synthetic methods of polypeptides containing new heart of kidney imaging agents. METHODS AND RESULTS: Five new target chelators--2-N-(2'-s triphenylmethylacetyl) amino-(N'-2"-N",N"-diethylethylamine) phenylpropamide (MPNE), 2-N-(2'-s-triphenylmethyl acetyl) amino-(N'-2"-N",N"-dimethylethylamine) phenylpropamide (MPNM), 2-N-(2's-triphenylmethylacetyl) amino-3-methyl-(N'-2" N",N"-dimethylethylamine) butyramide (MVNM), 2-N-(2'-s-triphenyl methylacetyl) amino-3-methyl-(N'-2"-N",N"-diethylethylamine) butyramide (MVNE), 2-N-(2'-s triphenylmethylacetyl) amino-(N'-acetylglycine) phenylpropamide (MPG2)--were synthesized through five steps with mercaptoacetic acid as primitive materials, all of which were identified on the basis of spectroscopic data, such as IR, 1HNMR, MS or elementary analysis. The protection of the mercapto group was improved and the relatively new reaction condition of active ester with amino acid is developed. All the chelators were labeled with Technetium-99m and their biological activities in mice given in ID values was tested to explore new heart imaging agents, where ID is the percentage injected dose per organ. The ID was determined by in vivo biodistribution study. Tc-99m complexes 0.1 mL was injected into the laterial tail vein of 3 anaesthetised rats. At 2, 5, 10, 30, 60 min post injection, rats were sacrificed by decapitation, bled from the neck and dissected. Organs were removed at dissection. The radioactivities in various organs were determined in an automatic twin crystal gamma counter. CONCLUSION: The bio-distribution results in mice indicate that 99Tcm-MVNM have higher heart uptake (ID = 8.40%/g, 2 min post-injection) and quicker blood clearance (ID = 4.3%/g, 60 min post-injection); 99Tcm-MPNE and 99Tcm-MPNM also have fairly high heart uptake and quick blood clearance; 99Tcm-MPG2 has better kidney accumulation and higher activity ratios of kidney to blood (about 4). PMID- 12579799 TI - [Study of the interactions between diorganotin (IV) complexes of 1,3-dimethyl-4 acetyl-5-pyrazolone and mononucleotides and DNA]. AB - AIM: The interactions between diorganotin (IV) complexes of 1,3-dimethyl-4-acetyl 5-pyrazolone (HL1) and mono-nucleotides together with DNA near physiological condition were investigated. METHODS: The mode of action of the diorganotin (IV) complexes with mononucleotides and DNA under different conditions and different times were investigated by high resolution NMR technology and UV spectra. RESULTS: The interaction of [(L1)2SnEt2] with AMP was shown to result in significant change of chemical shift of H(8), H(2) and 31P of AMP. Hyperchromic effect of DNA could be observed due to the interaction of; [(L1)2SnEt2] with DNA, while interaction of [(L1)2SnMe2] with AMP and DNA could only cause obvious change of chemical shift of 31P and lead to hypochromic effect of DNA. CONCLUSION: The results indicate that [(L1)2SnEt2] can selectively bind to the N1 atom of the base and the phosphate oxygen atom of AMP and may further destroy the helical structure of DNA, while the dimethyltin (IV) compound of 1,3-dimethyl-4 acetyl-5-pyrazolone [(L1)2SnMe2] merely binds to the the phosphate oxygen atom of AMP and causes the contraction of DNA helical structure. PMID- 12579801 TI - [Studies on the chemistry of two new compounds of Hemistepta lyrata]. AB - AIM: To study the chemical constituents of Hemistepta lyrata. METHODS: To separate compounds with various chromatography technology and to elucidate their structures by chemical and spectral analysis. RESULTS: Two compounds were isolated from Hemistepta lyrata and their structures were determined as hemiceramide (I), hemisterpene ether (II). CONCLUSION: Compounds I and II are new compounds. PMID- 12579800 TI - [Studies on the chemical constituents of Lycianthes biflora]. AB - AIM: To study the chemical constituents of Lycianthes biflora. METHODS: Column chromatography was used to separate the chemical constituents. IR, MS, 1HNMR, 13CNMR and 2D-NMR technique were used to determine the structures of the isolated constituents. RESULTS: Five compounds were isolated from this plant. Their structures were identified to be bifloride A (1), N-trans-cinnamoyltyramine (2), liquiritigenin (3), N-trans-p-coumaroyloctopamine (4), 1-O-beta-D-glucopyranosyl 2-N-2'-hydroxypalmitoyl-sphinga-4- trans-8-trans-dienine (5). CONCLUSION: Compounds 1 and 2 are new compounds, the others were isolated from this plant for the first time. Compound 2 showed inhibitory effects on P-388. PMID- 12579802 TI - [Analysis of the diterpenoids in the extract of Pteris semipinnata L by HPLC-APCI MS]. AB - AIM: To establish an accurate and reliable method for quantitative analysis of the diterpenoids in Pteris semipinnata L. METHODS: A quadruple mass spectrometer coupled with atmospheric pressure chemical ionization interface was employed as a detector for HPLC. As to MS detector, selective ion monitoring (SIM) scan mode was used. For ent-11 alpha-hydroxy-15-oxo-kaur-16-en-19-olic acid (5F) and ent-11 alpha-hydroxy-15-oxo-kaur-16(R) methyl-19-olic acid (4F), the majority of the diterpenoids in Pteris semipinnata L, the [M-H]-1 ion were observed, and the [M H2O-H]-1 ion could be observed from the collision-induced dissociation spectua. [M-H]-1 was selected as the SIM ion in quantification, the mobile phase and the MS conditions were optimized. The mobile phase of HPLC was 30% CH3CN-70% 2 mmol.L 1 NH4Ac, analytical column was Diamonsil ODS (4.6 mm x 150 mm), flow rate 1.0 mL.min-1, inject volume 5 microL. The area of ion flow peak were used for quantitative determination. As an example of its application, this method was used to determine the content of 5F as an antitumor diterpenoid in Pteris semipinnata L. RESULTS: The content of 5F accounted 1.18 mg.g-1 in Pteris semipinnata L sample. For 5F, RT is about 4.3 min, the standard curve showed good linearity over the range of 0.05-2.5 micrograms, gamma = 0.9998 (n = 5); the recovery was 97.8% (n = 5); the limit of detection was 0.4 ng (inject 5 microL). CONCLUSION: This method is highly sensitive, accurate and fast, which can be applied to study the antitumor drug of diterpenoids in Pteris semipinnata L and to establish the raw herb standard. PMID- 12579803 TI - [Studies on NaNO2-induced DNA mutation by convolution spectrometry]. AB - AIM: To study the sodium nitrite induced DNA mutation by convolution spectrometry (CS). METHODS: The spectra of sodium nitrite-induced mutative calf thymus DNA was compared with ego criteria based on Spectra of the primary DNA within the wavelength range from 200 to 340 nm. Distilled water served as blank and normal saline served as negative control. Any difference was quantitatively expressed by differential value (delta) of convolution spectra. Near-infrared spectroscopy was employed as the reference method. RESULTS: The differential value was positively correlated with the increasing time and concentration of sodium nitrite. delta values increased to 1.37%, 2.41% and 5.44% respectively within 2-hour's reaction between calf thymus DNA and 0.5, 0.05 and 0.005 microgram.mL-1 sodium nitrite, while on the contrary, changes could be hardly observed on the corresponding UV absorption spectra. The results were also confirmed by their corresponding near infrared spectra. CONCLUSION: The delta values can be used to represent the compound's strength of mutagenesis. Every convolution procedure takes less than one minute, so CS provides a fast, simple and inexpensive alternative method to determine chemical or medicinal DNA mutation. PMID- 12579804 TI - [Chiral separation and preparation of three new antagonists of alpha 1 adrenoceptors by chiral mobile phase HPLC]. AB - AIM: To establish new methods for the chiral separation and preparation of three new drugs, alfuzosin, terazosin and doxazosin. METHODS: By optimizing factors which affect the chiral separation, modifier of solvent, chiral additive, pH of mobile phase, modifier of organic base and stationary phase, the optimum condition for chiral separation were selected. The preparation of enantiomers was carried out on semi-preparative reverse phase column (7.8 mm x 250 mm C4 5 microns). Acetonitrile-water modified by the addition of carboxymethyl-beta cyclodextrin (2%-5%, w/v) was applied as chiral mobile phase. RESULTS: The enantiomers of three new drugs were base-line-separated and milligram-scale samples of enantiomer were obtained. CONCLUSION: The newly established method can be used in research and development of the enantiomers of three new drugs. PMID- 12579805 TI - [Electrochemical behavior of dopamine at the poly (isonicotinic acid) modified glassy carbon electrode]. AB - AIM: To study the determination of dopamine (DA) in the presence of ascorbic acid (AA) using poly (isonicotinic acid) film modified electrode. METHODS: The cyclic voltammetry and differential pulse voltammetry were used to study the electrochemical behavior of DA at the poly (isonicotinic acid) film modified electrode. RESULTS: The poly (isonicotinic acid) film modified electrode showed an electrocatalytic effect on DA, and shifted the oxidation of AA to negative potential. The difference between the oxidation potentials of DA and AA was 204 mV, thus, AA did not interfere with the determination of DA. The linear range between the anodic currents and DA concentration was: 1.0 x 10(-7)-2.0 x 10(-5) and 2.0 x 10(-5)-1.0 x 10(-4) mol.L-1. The detection limit was 8.0 x 10(-9) mol.L 1. CONCLUSION: The useful life period of the modified electrode is three weeks at least. The modified electrode can be used to the determination of DA in the sample. PMID- 12579806 TI - [RP-HPLC determination of diphenytriazol in rat liver microsomal incubates and its application in in vitro metabolism]. AB - AIM: To establish a RP-HPLC method for determination of diphenytriazol (DL111-IT) in rat hepatic microsomes. METHODS: DL111-IT in rat hepatic microsomal incubates was extracted with chloroform, using diazepam as internal standard. The determination was performed on a Lichrospher ODS-C18 reversed column (25 cm x 0.46 cm ID) with mobile phase of methanol-pH 7.5 phosphate buffer (70:30) at a flow-rate of 1.0 mL.min-1. A UVVIS detector was operated at 235 nm. RESULTS: The assaywas linear from 1.01-101.0 micrograms.mL-1 for DL111-IT. The limit of detection was 0.15 microgram.mL-1 (signal-to-noise ratio 3) and the limit of quantification was 1.01 micrograms.mL-1(RSD < 10%, n = 4). The method afforded average recoveries of (100.3 +/- 1.9)% (n = 5), and intra-day and inter-day RSD were less than 5.0%(n = 5). The method allowed study of the in vitro phase I metabolism of DL111-IT in rat liver microsomal incubates. The microsomes induced by beta-naphthoflavone showed high enzymatic activity for DL111-IT phase I metabolism. CONCLUSION: The method is simple, accurate and can be used to study the metabolism of DL111-IT in rat hepatic microsomes. PMID- 12579807 TI - [Impurity analysis and their structure determination of gatifloxacin]. AB - AIM: To analyse the impurities of gatifloxacin. METHODS: The impurity of gatifloxacin were analysized and determinated by RP-HPLC/electrospray ionization mass spectrometry with a Zorbax SB-C18(4.6 mm x 150 mm ID, 5 microns). The mobile phase was 3% acetic acid/acetonitrile-3% acetic acid/water (15:85). The two compounds were synthesized: 1-cyclopropyl-6-fluoro-1, 4-dihydro-8-methoxy-7-(1 piperazinyl)-4-oxo-3-quinolinecarboxylic acid (DMP) and 1-cyclopropyl-6-fluoro-1, 4-dihydro-8-hydro-7-(3-methy-1-piperazinyl)-4-oxo-3-quinolinecarboxylic acid (DMO). Their liquid chromatogram, UV, MS were compared with those of the impurity of gatifloxacin. RESULTS: The mass of the impurity was 14 less than that of gatifloxacin. It means the impurity was CH2 less than gatifloxacin. The tR (HPLC), UV and MS of DMP were the same as those of the impurity of gatifloxacin. CONCLUSION: Based on the tR (HPLC), UV and MS, the impurity of gatifloxacin is confirmed as DMP. PMID- 12579808 TI - [Preparation of long circulating mitoxantrone liposomes and its pharmacokinetics]. AB - AIM: To prepare the mitoxantrone long circulating liposomes (LCL) and to observe the residence behavior of the mitoxantrone LCL in rabbits. METHODS: The long circulating mitoxantrone liposomes were prepared by ethanol injection combined with the ammonium sulphate gradients method. Amphipathic polyethylene glycol distearoyl phosphatidylethanolamine (PEG-DSPE) was added to modify the membrane of the liposomes. The column-switching system of RP-HPLC was utilized to determine mitoxantrone concentration in rabbit plasma. RESULTS: The mean diameter of the long circulating mitoxantrone liposomes was 60 nm, with the entrapping efficiency of 93.6%. With the same dosage (2 mg.kg-1) i.v. in rabbits, the mean residence time (MRT) of the long circulating mitoxantrone liposomes was 9.8 h, while that of the normal liposomes was 3.6 h, and the AUC of the former is 6.4 fold greater than of the latter. It showed that the long circulating mitoxantrone liposomes prolonged the resident time of the drug in the blood circulating system and they reduced the uptake by the reticuloendothelial system, simultaneously. CONCLUSION: Liposomes with high entrapping efficiency and small particle size could be prepared by ethanol injection combined with the ammonium sulphate gradients method, and the liposomes modified by PEG-DSPE could raise the AUC and prolonged the resident time of the drug in the blood circulating system. PMID- 12579809 TI - [Effect of solution viscosity on polymer precorneal residence time evaluated by in vitro method]. AB - AIM: To evaluate how solution viscosity affects the precorneal residence of five water-soluble polymers with different properties. METHODS: Captive bubble technique was used, with the consecutive change of contact angle interpreted as an indication of desorption process, to study the residence of those polymers in vitro on freshly enucleated rabbit eyes under physiological conditions. RESULTS: Carbopol and sodium hyaluronate (HA), which adsorbed to isolated ocular surface more than 15 min, showed the optimum precorneal retentive capabilities. When the solution viscosity increased from 12 mPa.s to 50 mPa.s, the residence time of carbopol and HA were prolonged 10 min and 7 min, respectively, but that of sodium carboxymethylcellulose was not affected. CONCLUSION: The result suggested that higher viscosity is beneficial to improve the ocular residence time of bio adhesive polymers. PMID- 12579810 TI - [Preparation and characterization of prednisolone-poly (hydroxybutyrate-co hydroxyvalerate) nanoparticles]. AB - AIM: To optimize the preparation of sustained release prednisolone-poly (hydroxybutyrate-cohydroxyvalerate) (PNS-PHBV) nanospheres (NP) using the novel biodegradable materials PHBV as the carriers and PNS as a model drug. METHODS: PNS-PHBV nanospheres were prepared by ultrasonic-emulsion technique. The diameter, its distribution and Zeta potential on the surface of particles were measured by means of Zetasizer. RESULTS: The diameter of NP is in the range of 50 250 nm. The drug loading of NP increases but incorporation efficiency and Zeta potential dramatically decrease with increasing ratio of the feeding quantities of drug to those of carriers. The drug release behavior in vitro appeared to have biphasic characteristics with initial burst effect. The more burst effect, the less the diameters of nanoparticles. The longest release time was up to 32 h. CONCLUSION: The technology of preparation is reasonable and PNS-PHBV nanoparticle showed significant sustained release. PMID- 12579811 TI - [The anticancer effect of artesunate and its mechanism]. AB - AIM: To study the anticancer effect of artesunate and its mechanism. METHODS: To observe the effect of artesunate on the growth of solid tumor and the expression of PCNA, Bcl-2 and Bax genes in mice bearing H22 solid hepatic carcinoma. RESULTS: After administration of artesunate (ig, 300 mg.kg-1.d-1 x 7 d), growth of solid tumor was obviously inhibited, the tumor inhibitory rates were 49.1%, 48.7% and 46.6% and 46.6% in 3 experiments. The apoptosis of liver cancer cells were increased. Immunohistochemical staining showed that the number of Bcl-2 protein positive cells were decreased, but the number of Bax protein positive cells were increased. The PCNA positive cells were significantly lower than those in the control group. CONCLUSION: Artesunate showed obvious anticancer activity on H22 hepatic carcinoma bearing mice and undergo apoptosis of liver cancer cells. The mechanism of anticancer effect of artesunate may be related to down regulation of the expression of PCNA and Bcl-2 genes and up-regulation of the expression of Bax gene. PMID- 12579812 TI - Advance of immunoaffinity capillary electrophoresis. PMID- 12579813 TI - [Application of liquid chromatography-spectroscopy coupled techniques to the chemical screening of natural products]. PMID- 12579814 TI - [Application of genomics and biochip technology in research and development of chinese materia medica]. PMID- 12579815 TI - [mRNA expressions of voltage-dependent potassium channels in the brain of scopolamine-induced memory impaired rats]. AB - AIM: To study mRNA expression difference of voltage-dependent potassium channels in the brain of scopolamine-induced memory impaired rats. METHODS: Memory impairments induced in rats by scopolamine (1 mg.kg-1) were assessed in the Morris water maze test. After rats were injected intraperitoneally with scopolamine for 6 days, the mRNA expression level of five voltage-dependent potassium channels, Kv1.4, Kv1.5, Kv2.1, Kv4.2 and Kv4.3 were detected in the rat cortex and hippocampus by RT-PCR. RESULTS: Scopolamine (1 mg.kg-1) was shown to significantly induce memory impairment in rats. The mRNA levels of Kv4.2 were decreased by 28.8% and 33.9% in the cortex and hippocampus, respectively. The mRNA levels of Kv1.4 and Kv2.1 were increased in the hippocampus by 111.7% and 64.3%, respectively. There were no differences in the brain mRNA levels of other voltage-dependent potassium channels in scopolamine-induced memory impaired rat. CONCLUSION: The mRNA expression levels of voltage-dependent potassium channels changed significantly in the brain of scopolamine-induced memory impaired rats. PMID- 12579816 TI - [Experimental studies on the anti-thrombosis effect of 3,4-oxo-isopropylidene shikimic acid]. AB - AIM: To study the effect of 3,4-oxo-isopropylidene-shikimic acid (ISA) against arteriovenous shunt and middle cerebral artery thrombosis (MCAT) in rats. METHODS: Arteriovenous shunt model was adopted to measure thrombus weight; The neurologic deficit (ND) and the infarct size (IS) of the middle cerebral thrombosis (MCAT) model induced by FeCl3 were observed; The effect of ISA on platelet aggregation rate of rat and rabbit by giving ISA in vivo and in vitro was studied. RESULTS: ISA 25, 50, 100 and 200 mg.kg-1 ig was shown to reduced the weight of thrombus in arteriovenous shunt model; ISA 50, 100 and 200 mg.kg-1 ig for 2 times in 24 hours, attenuated the ND of rats subjected to MCAT; ISA 100 and 200 mg.kg-1 reduced IS of rats after MCAT by 27.8% and 31.6%, respectively; ISA 50, 100 and 200 mg.kg-1 ig inhibited platelet aggregation of normal rats; ISA 10( 3)-10(-5) mol.L-1, inhibited rabbit platelet aggregation in vitro. CONCLUSION: ISA inhibited thrombosis by anti-platelet-aggregation. PMID- 12579817 TI - [Possible mechanisms of the protective effect of ginsenoside Rg1 on apoptosis in substantia nigra neurons]. AB - AIM: To investigate the role of ginsenoside Rg1 in preventing against 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced apoptosis of the substantia nigra neurons in the mouse model of Parkinson's disease (PD). METHODS: C57B1 male mice were given MPTP i.p. in the PD model group. Different doses of ginsenoside Rg1 (2.5, 5.0 and 10.0 mg.kg-1) were given i.p. 3 days prior to MPTP in the pretreatment group. Nissl staining, tyrosine hydroxylase (TH) immunostaining and TdT-mediated duTP nick-end labeling (TUNEL) staining were used to observe the damage of nigral neurons. The method of immunostaining was used to detect the caspase-3 activity, expression of inducible nitric oxide synthase (iNOS) and neuronal nitric oxide synthase (nNOS). RESULTS: Pretreatment with ginsenoside Rg1 was shown to prevent the loss of Nissl staining neurons and TH-positive neurons, and decrease the percent of TUNEL-positive. Simultaneously, Rg1 was found to reduce caspase-3 activity and the expression of iNOS. CONCLUSION: Ginsenoside Rg1 showed protective effect on MPTP-induced apoptosis in the mouse nigral neurons and this effect may be attributable to reducing the expression of iNOS and inhibiting the activation of caspase-3. PMID- 12579818 TI - [Studies on the inhibition of polyhydroxylated aromatic compounds against HIV-1 integrase]. AB - AIM: Three major enzymes of HIV-1, reverse transcriptase (RT), protease (PR), and integrase (IN), are important targets for anti-HIV drugs. Nine RT and five PR inhibitors have been effectively used in treatment of AIDS patients. In order to find active integrase inhibitors, twenty polyhydroxylated aromatic compounds were tested. METHODS: ELISA method was used to test the integrase activity. The synthesized donor substrate oligonucleotide representing the HIV-1 U5LTR was immobilized onto Covalink polystyrene microtiter plates, and a synthesized biotinlated 20 bp oligonucleotide was used as the target substrate. The products were detected and quantified by a colorimetric avidin-linked alkaline phosphatase reporter system. RESULTS: Compound NQ-2 was found to inhibit HIV-1 integrase with the IC50 of 78.5 mumol.L-1 by ELISA method. Its novel analogue NQ-3 was found to be 2 fold more potent on HIV intrgrase than NQ-2, IC50 was 37.2 mumol.L-1. The IC50s of NQ-2 and NQ-3 to inhibit the 3'-pro + assembly activity of integrase were 96.94 mumol.L-1 and 8.48 mumol.L-1; to inhibit assembly activity were 168 and 6.9 mumol.L-1 and to inhibit strand-transfer activity were 49.8 and 1.1 mumol.L-1, respectively. Compound NQ-2 mostly inhibited the strand transfer activity of HIV-1 integrase. Compound NQ-3 inhibited both the assembly and strand transfer with high activities. CONCLUSION: Naphthoquinone compound NQ-3 was found to be a novel HIV integrase inhibitor which warrants further study. Uncoupled ELISA HIV integrase assay is shown to be useful to screen HIV-1 integrase inhibitors. PMID- 12579819 TI - [Three dimensional quantitative structure-activity relationship of a series of benzocylohepatpyridine farnesyltransferase inhibitors]. AB - AIM: To build a three dimensional structure model that correlates the biological activities and the structures of a series of 1-(8-chloro-6,11-dihydro-5H benzo[5,6]cyclohepta-[1,2-]pyridin-11-yl) piperazines farnesyl protein transferase (FPTase) inhibitors. METHODS AND RESULTS: Mutation in the ras oncogene takes place in many human cancers, involving 30%-50% of colon and 90% of pancreatic cancer. Ras proteins function as central switches for signals given by growth factors that direct cell growth and cell differentiation. The dependence of the transforming activity of Ras on the farnesylation has led to intense search for FPTase inhibitors that may have therapeutic pontetial as anticancer agents. This paper is to build a three dimensional structural model that correlates the biological activities and the structures of a series of FPTase inhibitors. The investigated sixty-nine inhibitors contain six types of structures, the optimal conformations of which were studied using system search. A three dimensional quantitative structure-activity relationship (3D-QSAR) model was constructed using the method of comparative molecular field analysis (CoMFA). The resulting cross-validation R2 is 0.581, non-cross-validation R2 0.968, SE 0.148, F 198.7. The predicted activities of 10 inhibitors using this 3D-QSAR model are comparable to the experimental activities, indicating that the 3D-QSAR model has ability to predict activities of new inhibitors and offers an approach to design new FPTase inhibitors. CONCLUSION: The information of CoMFA model offers an approach to designing new FPTase inhibitors. PMID- 12579820 TI - [Chemical composition of fructus Liquidambaris--lulutong]. AB - AIM: To study the chemical composition in the fruits of Liquidambar formana Hance. METHODS: Various column chromatographic techniques were used to separate and purify the constituents. Their physical and chemical properties and spectral data were measured for structural elucidation. RESULTS: Eleven compounds were isolated and identified as beta-sitosterol (1), 3-oxo-11 alpha, 12 alpha epoxyleanan-28, 13 beta-olide (2), 3-oxo-12 alpha-hydroxy-oleanan-28, 13 beta olide (3), 3 alpha-acetyloxy-25-hydroxyolean-12-en-28-oic acid (4), oleanolic acid (5), ursolic acid (6), daucosterol (7), betulonic acid (8), gallic acid (9), nonacosane (10) and n-triacontanoic acid (11). CONCLUSION: Among the isolated constituents, compound 4 is new compound, compound 3 is firstly isolated from the natural product and compound 5, 6, 7, 9, 10 and 11 are isolated from LuLuTong for the first time. PMID- 12579821 TI - [Isolation and identification of steroidal saponins in total saponin from Dioscorea nipponica Makino]. AB - AIM: To investigate the water-soluble steroidal saponins in total saponin from Dioscorea nipponica Makino and look for new active compounds. METHODS: The compounds were isolated with silica gel, PTLC and HPLC, and their structures were elucidated by acid hydrolysis, physical and chemical data and spectral analysis (IR, NMR, MS, HMQC, HMBC) as well as chemical correlations. RESULTS: The two steroidal saponins (water-insoluble saponin and water-soluble saponin) were isolated from the total saponin of Dioscorea nipponica Makino. The structures were elucidated as diosgenin 3-O-[alpha-L-rhamnopy ranosyl (1-->2)-[beta-D glucopyranosyl(1-->3)]]-beta-D-glucopyranoside (I), diosgenin 3-O-[alpha-L rhamnopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl (1-->4)]-beta-D-glucopyranoside (II). CONCLUSION: Compound II is a new steroidal saponin and firstly isolated from Dioscorea nipponica Makino. It was named as dioscin Dc. PMID- 12579822 TI - [Chemical constituents from marine fungus Penicillium thomii]. AB - AIM: To investigate the bioactive constituents from the mycelium of Penicillium thomii. Which isolated from Anemone collected in Qingdao beach. METHODS: The constituents were separated by using various chromatography and the structures were identified on the basis of extensive spectral analysis. RESULTS: Five compounds, namely penicillixanthone A (I), p-methylbenzolic acid (II), 1-O hexadecanoyl-2-O-(9-octadecenoyl)-3-O-(9, 12-octadecadienoyl) glycerol (III), 5 alpha, 8 alpha-epidioxy-24 zeta-methylcholesta-6, 22-dien-3 beta-ol (IV) and 1, 6, 8-trihydroxyl-3-methyl-9, 10-anthracenedione (V), were isolated from the mycelium of Penicillium thomii. CONCLUSION: Penicillixanthone A is a new compound, while the others are isolated from Penicillium thomii for the first time. PMID- 12579823 TI - [Approach to the crystalline characteristics of ceftezole sodium]. AB - AIM: To study the crystalline characteristics of ceftezole sodium. METHODS: Ceftezole sodium crystals were obtained from different solvents. X-ray diffraction, DSC, TGA, etc were used to analyze the crytals. RESULTS: Ceftezole sodium crystal was easily obtained in isopropanol-water mixture. It consists of ceftizole sodium monohydrate, which consists of type I and type II two different crystal forms. Powder X-ray diffraction patterns showed differences between type I and the type II crystal forms. Peaks at 8 degrees and 18 degrees in diffractograms of the type I, but at 9 degrees and 18.6 degrees in the type II could be observed. Water molecules in different crystal forms had different combining condition. They lost during 35-117 degrees C in the type I form, but lost during 110-160 degrees C in the type II form. CONCLUSION: Structure of ceftizole sodium monohydrate crystal obtained in different circumstance could be some vary, which influence upon the thermal stability of the compound. The type I crystal form is more stable than the type II. PMID- 12579824 TI - Determination of flavonoids in Hypericum perforatum by HPLC analysis. AB - AIM: To establish an HPLC method for simultaneous determination of four flavonoids in Hypericum perforatum L., ruitn, hyperin, avicularin and quercetin. METHODS: C18 column was used, the chromatography was carried out with a linear gradient program. The mobile phase was A: H2O (pH 3.0-3.5 adjusted with phosphoric acid) and B: acetonitrile, at flow rate of 1.0 mL.min-1, peaks were detected at 254 nm. RESULTS: The linear range of rutin was 1.376-8.256 micrograms.mL-1 (gamma = 0.9999), hyperin 3.160-18.960 micrograms.mL-1 (gamma = 0.9996), avicularin 0.968-5.808 micrograms.mL-1 (gamma = 0.9998) and quercetin 0.776-4.656 micrograms.mL-1 (gamma = 0.9993). The average recovery of rutin was 97.8%, RSD 4.8% (n = 3), hyperin 100.7%, RSD 3.7% (n = 3), avicularin 97.3%, RSD 0.8% (n = 3) and quercetin 100.5%, RSD 4.4% (n = 3). All of RSDs of precision were less than 2% (n = 5), reproducibilities less than 3%. CONCLUSION: The method is simple, effective and feasible. It can be used to determine flavonodis in Hypericum perforatum. PMID- 12579825 TI - [Study on the metabolites of guanfu base A hydrochloride in rat urine by high performance liquid chromatograph-mass spectrum]. AB - AIM: To study the metabolites of guanfu base A hydrochloride (GFA) in rat urine. METHODS: Rat urine was collected after i.v. injection of GFA. Phase I metabolites were identified by HPLC/MS and by comparison with authentic standards. Phase II conjugates were treated with either glucuronidase or sulfatase in presence or absence of glucuronidase specific inhibitor D-saccharic acid beta-1,4-lactone. The aglycones were identified by HPLC/MS. RESULTS: Phase I metabolites guanfu base I (GFI) and guanfu alcohol-amine (AA) were separated and identified in rat urine by comparison with authentic standards. Phase II conjugates, for which no authentic standards were available, GFA glucuronide and sulfate conjugates, GFI glucuronide and sulfate were separated and tentatively identified by hydrolysis with glucuronidase or sulfatase, the aglycones, GFA and GFI, were identified in rat urine. CONCLUSION: After i.v. injection of GFA, GFA is metabolized into GFI, AA, GFA glucuronide and sulfate conjugates, GFI glucuronide and sulfate conjugates in rat urine. The polarity of the metabolites is increased, and the effectiveness of them is lower than the parent drug. PMID- 12579826 TI - [The metabolic transformation of (-)-securinine]. AB - AIM: To study the in vitro and in vivo metabolism of (-)-securinine. METHODS: The metabolic transformation of (-)-securinine was studied by using phenobarbital induced rat liver microsomal incubate containing the NADPH-generating system in vitro and the constitution of the system was optimized. A reversed phase HPLC method was established to analyze the parent drug and its metabolites. The major metabolites were isolated and purified by liquid-liquid extraction, preparative TLC and HPLC, and their structures were elucidated as 6-hydroxyl securinine, 6 carbonyl securinine, 5 beta-hydroxyl securinine and 5 alpha-hydroxyl securinine by 1HNMR, 13CNMR and MS spectral analysis. An HPLC method was developed to analyze securinine and its metabolites in biofluids (bile, urine) of rat. The bile, urine and their enzymatic hydrolyzed samples of the rat i.p. administrated with (-)-securinine were determined by using this method. RESULTS: Four main metabolites of (-)-securinine in rat hepatic microsome incubation were obtained and their structures were elucidated. Metabolites from in vitro study were confirmed in biofluids (bile, urine) which were collected from rats given securinine i.p. It was suggested that 6-hydroxyl securinine was excreted in conjugated form as well by analyzing enzymatic hydrolyzed bile. CONCLUSION: The main metabolic pathway of (-)-securinine in vitro and in vivo is basically elucidated. PMID- 12579827 TI - [Relativities between lattice changes and the function of dissolution improvement of poorly soluble drug silymarin based upon PEG 6,000 solid dispersion system]. AB - AIM: To investigate the lattice mechanisms involved in the increased dissolution effect of polyethylene glycol (PEG 6,000) dispersion system on poorly soluble drug silymarin (SILY). METHODS: Fusion method was used to prepare the solid dispersions of SILY with PEG 6,000. Evaluation of the improvement of dissolution was performed with dissolution studies in vitro. X-ray powder diffraction combined with diffraction peak pattern-fitting procedure were applied to quantitatively analyze the changes of lattice parameters. The interaction of SILY and PEG 6,000 was also determined with Fourier transform-infrared (FT-IR) spectroscopy. RESULTS: The dissolution rate of SILY was considerably increased when formulated in solid dispersion of PEG 6,000 as compared to pure SILY. The datum from the X-ray diffraction showed the changes in the lattic spacings and particular diffraction peaks (position and the intensity) of PEG 6,000 and SILY. These could explain the increased rate of SILY released from solid dispersion system. The information of FT-IR spectroscopy showed the absence of well-defined drug-polymer interaction. CONCLUSION: The dissolution improvement of poorly soluble SILY from solid dispersion of PEG 6,000 can be illuminated by the changes of the lattice parameters of PEG 6,000 and the drug. PMID- 12579828 TI - [Study on pharmaceutical characterization and pharmacokinetics of daunorubicin long-circulating liposomes in rat]. AB - AIM: To study the pharmaceutical characterization and pharmacokinetics of long circulating liposomes containing daunorubicin. METHODS: The morphology of daunorubicin long-circulating liposome was surveyed under the transmission electron microscope. The size of daunorubicin long-circulating liposomes was determined by laser scatter method. The entrapment efficiency and accelerative experiment stability of the daunorubicin long-circulating liposomes were examined. Visible spectrophotometry and the HPLC method were established for determination of the daunorubicin in the long-circulating liposomes. The percent release of daunorubicin from long-circulating liposomes in HBS (pH 7.5) and rat serum at 37 degrees C were examined. The pharmacokinetics in rats were studied. RESULTS: The high entrapment efficiency (> 85%) and stabilized long-circulating liposomes could be achieved. The drug was slowly released from the daunorubicin long-circulating liposomes. The drug released from liposomes was less than 10% in 24 h. The T1/2 alpha and AUC of long-circulating liposome were higher than those in injections. CONCLUSION: The long-circulating liposomes prepared by us have high encapsulation efficiency and the pharmaceutical characterization showed good stability, they can be used for clinical purpose. PMID- 12579829 TI - [DNA profiling of Pogostemon cablin chemotypes differing in essential oil composition]. AB - AIM: To provide molecular evidence for quality evaluation and GAP production of Pogostemon cablin (Blanco) Benth. cultivated in different regions in Guangdong and Hainan provinces, China, by comparing two sequences (1.2 kb of plastid matK gene and 1.8 kb of nuclear 18S rRNA gene) and two chemotypes (Pogostone-type and Patchouliol-type in essential oil composition). METHODS: PCR direct sequencing was applied to detemine the matK and 18S rRNA sequences for six samples of Pogostemon cablin from different localities. RESULTS: The matK sequences of six samples of Pogostemon cablin from different regions of cultivation are 1,245 bp in length, which coding 415 amino acids of protein (maturase), and 18S rRNA sequences are 1,803-1,805 bp in size. Based on multiple sequence alignment, there are 47 variable sites in the matK sequence of these six samples, 17 in the 18S rRNA sequence. The cluster tree reconstructed by UPGMA method shows that the sequence divergence both in matK and 18S rRNA genes among six samples of Pogostemon cablin was well correlative with their regions of cultivation and intraspecific chemotypes of essential oil composition. CONCLUSION: Combining with chemical and biogeographical data, DNA sequencing can become a powerful tool in the key technique-species identification of quality evaluation and GAP production of Pogostemon cablin. PMID- 12579830 TI - [Advances in the research on nicotinic acetylcholine receptors agonists]. PMID- 12579831 TI - [Application of microdialysis in pharmacokinetic and drug metabolism studies]. PMID- 12579832 TI - [Establishment of immune insulin resistance model in the rats by i.v. injection of BCG]. AB - AIM: To observe the stability of BCG-induced insulin resistance model. METHODS: The glucose tolerance, serum glucose, FFA, insulin, triglycerides, cholesterol, TNF-alpha and ALT level were measured. The change of GDR was measured by euglycemic clamp in model rats after given i.v. BCG 2, 4 and 8 weeks. RESULTS: After 2, 4 and 8 weeks, the GIR and glucose tolerance of the animals deceased significantly. After 2, 4 and 8 weeks, BCG infusion resulted in a pronounced reduction in glucose tolerance and insulin-stimulated glucose disposal rate [GDR = GDR: (29 +/- 6) vs (13 +/- 7) mg.kg-1.min-1 2 weeks; (29 +/- 6) vs (11 +/- 7) mg.kg-1.min-1 4 weeks and (23 +/- 3) vs (16 +/- 3) mg.kg-1.min-1 8 weeks, respectively, P < 0.01]. BCG infusion resulted in a pronounced increase in the weights of the liver [(6.2 +/- 0.9) vs (8.2 +/- 1.3) g, P < 0.05] and spleens [(0.51 +/- 0.11) vs (1.4 +/- 0.4) g, P < 0.01]. The histo-pathological results showed that BCG infusion resulted severe inflammation in the livers and spleens and the ratio of beta/alpha in pancreas increased. The serum levels of triglyceride, FFA and glucose were unchanged, but the level of serum TNF-alpha [543 +/- 60) vs (759 +/- 137) pg.mL-1, P < 0.05] and insulin [(31 +/- 5) vs (36 +/- 5) mu.L-1, P > 0.05] increased. CONCLUSION: This novel model of immune insulin resistance is completely and constantly established. PMID- 12579833 TI - [Effect of recombinant prothymosin alpha on secretion of IFN-gamma, IFN-alpha and TNF-alpha in vitro]. AB - AIM: To study the effect of prothymosin alpha (Pro T alpha) as a fusion protein on secretion of IFN-gamma, IFN-alpha and TNF-alpha in vitro. METHODS: The in vitro study was carried out on the culture of splenocytes, splenic and peritoneal macrophages isolated from Balb/c mice. Splenocytes were incubated with various concentrations of Pro T alpha (1 x 10(-7)-1 x 10(-10) mol.L-1) with or without Con A (5 micrograms.mL-1) for 72 h. Splenic and peritoneal macrophages were respectively treated with Pro T alpha (1 x 10(-7)-1 x 10(-10) mol.L-1) in the presence of LPS (10 micrograms.mL-1) for 24 h. Then IFN-gamma, IFN-alpha and TNF alpha levels in the supernatant were detected by ELISA. RESULTS: Pro T alpha (1 x 10(-7) mol.L-1) was found to obviously increase IFN-gamma level (P < 0.05) in the supernatant of splenocytes compared with the control group. Moreover, Pro T alpha (1 x 10(-7) mol.L-1) significantly induced the secretion of IFN-alpha (P < 0.01) and TNF-alpha (P < 0.01) in splenic and peritoneal macrophages. CONCLUSION: In vitro, Pro T alpha could increase the secretion of IFN-gamma, IFN-alpha and TNF alpha. PMID- 12579834 TI - [The protective effects of melatonin on global cerebral ischemia-reperfusion injury in gerbils]. AB - AIM: To investigate the effects of melatonin (MT) on histology and behavioral tests during global cerebral ischemia-reperfusion in gerbils. METHODS: Global cerebral ischemia was induced by occluding the bilateral common carotid arteries for 10 min in gerbils. Three doses of MT were administrated intraperitoneally 30 min prior to the onset of ischemia. Locomotor activity was measured by using the open field method 3 and 7 days after the ischemic episode. T maze test was carried out 4, 5 and 6 days after ischemia to assess the working memory of gerbils. Neuronal damage was assessed in CA1 pyramidal layer of gerbil hippocampus and evaluated 7 days after ischemia. RESULTS: MT significantly reversed the locomotor activity increases, ameliorated learning and working memory deficit, and reduced the extent of CA1 hippocampal pyramidal cells injury after transient global cerebral ischemia in the Mongolian gerbil. CONCLUSION: MT provides significantly protective effect against both histological and behavioral consequences of global cerebral ischemia-reperfusion injury in gerbils. PMID- 12579835 TI - [FAK antisense oligodeoxynucleotides inhibit vascular smooth muscle cell migration and adhesion mediated by FAK-ERK1/2 signaling pathway]. AB - AIM: To study the effects of FAK-ERK1/2 signaling pathway and FAK antisense oligodeoxynucleotides (ODNs) on vascular smooth muscle cell (SMC) migration and adhesion stimulated by fibronectin (FN). METHODS: Migration and adhesion of cultured SMCs were stimulated by different concentrations of FN, FAK, ERK1/2. And their phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense ODNs were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation, SMCs migration and adhesion were also measured by modifing Boyden Chamber and morphological enumeration, respectively. RESULTS: FAK were expressed when SMCs adhesion and migration were successfully simulated by FN (5, 10, 20, 40, 60 micrograms.mL-1), high contents of FAK and ERK1/2 phosphorylation were detected by 20 micrograms.mL-1 FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid. FAK and ERK1/2 phosphorylation were inhibited magnificently after FAK antisense ODNs transfection. Cell migration stimulated by FN 10, 20, 40 and 60 micrograms.mL-1 were reduced by 23.26%, 21.63%, 19.31% and 17.88% respectively (P < 0.05). SMCs adhesive spreading in 5-60 micrograms.mL-1 FN groups were reduced by 17.89% 27.67% (P < 0.05). CONCLUSION: FAK-ERK1/2 mediated signal transduction play important roles in SMCs migration and adhesion stimulated by extracellular matrix. The process can be inhibited by FAK antisense ODNs effectively. PMID- 12579836 TI - [Protective effect of tripchlorolide on dopaminergic neurons in partially lesioned rat model of Parkinson's disease]. AB - AIM: To study whether the immunosuppressant tripchlorolide (T4) exerts neuroprotective effect on dopaminergic neurons. METHODS: A rat model of Parkinson's disease (PD) was set up by transection of the medial forebrain bundle (MFB) with a wire knife. The rotational behavior, HPLC-ECD, tyrosine hydroxylase (TH) immunocytochemistry, ELISA methods were used to evaluate the influence on the dopaminergic neurons following T4 treatment. RESULTS: T4 treatment was shown to effectively attenuate the rotational behavior challenged by amphetamine (2.5 mg.kg-1, i.p.) in the PD rats. T4 markedly prevented the decrease of dopamine content in the striatum and the loss of dopaminergic neurons in the substantia nigra pars compacta. T4 was found to suppress the abnormal increase of TNF-alpha and IL-2 level in brain tissues of PD rats after MFB transection. CONCLUSION: The evidence that the immunosuppressive Chinese herb extract T4 possesses neuroprotective activities on the dopaminergic neurons in PD rats was presented. The underlying mechanism of T4 may be relevant to its immunosuppressive activity. PMID- 12579837 TI - Synthesis of 1-(3-phthalimido-2-oxobutyl)-4-substituted- phenylpiperazines and their anti-HIV reverse transcriptase activity. AB - AIM: Synthesis of 1-(3-phthalimido-2-oxobutyl)-4-substituted- phenylpiperazines (5-15). METHODS: The starting material nitrogen mustard hydrochloride (16), reacted with the corresponding substituted anilines to afford piperazine hydrochlorides (17-27), which were then coupled with 1-bromo-3-phthalimidobutan-2 one (4) to give the target compounds. RESULTS: Eleven target compounds (5-15) were synthesized, which were characterized by 1HNMR, IR and elemental analysis. CONCLUSION: Anti-HIV-1 RT using HIV reverse transcriptase P-66 protein test showed that compounds 11, 14, 10 and 13 possessed inhibitory effects against HIV 1 reverse transcriptase (RT), with IC50 29.80, 35.20, 43.77 and 63.76 mumol.L-1, respectively. PMID- 12579838 TI - [Studies on the chemical constituents of Securidaca inappendiculata]. AB - AIM: To investigate the chemical constituents of Securidaca inappendiculata. METHODS: Compounds were isolated by silica gel column chromatography and Medium Pressure Liquid Chromatography, respectively. Structures of the compounds were elucidated by chemical evidence and spectral (UV, IR, MS, 1HNMR and 13CNMR) analysis. RESULTS: Eight compounds (1-8) were isolated and identified. Seven of them (2-7) are known compounds: oleanolic acid, p-hydroxytruxinic acid, p coumaric acid, ethyl p-methoxycinnamate, uridine, beta-sitosterol and daucosterol. One (1) is a new hemiterpenoic acid glycoside, named securiterpenoside. CONCLUSION: A new hemiterpenoic acid glycoside, named securiterpenoside, was isolated from Securidaca inappendiculata Hassk. Seven known compounds were isolated from genus Securidaca for the first time. PMID- 12579839 TI - [Studies on the chemical constituents of Pseudotsuga sinensis]. AB - AIM: To study the chemical constituents of Pseudotsuga sinensis Dode (Pinaceae). METHODS: To separate the constituents of P. sinensis by using various kinds of chromatography and identify their structures on the basis of spectral analysis. RESULTS: Six compounds were isolated from P. sinensis. Their structures were established as 5,7,4'-trihydroxy-6-methylflavanone (poriol, I), 3,5,7,3',4'- pentahydroxyflavonone (quercetin, II), 5,7,3',5'-tetrahydroxy-6- methylflavanone (III), 5,7,3',5'-tetrahydroxyflavanone (IV), 3,5,7,3',5'-pentahydroxyflavanone (V) and 5-hydroxy-6-methylchromone-7-O-beta-D-glucopyranoside (VI) based on the analysis of spectral data of IR, UV, MS, 1D and 2D-NMR. CONCLUSION: Compounds III and VI are new compounds. All of six compounds were isolated from this plant for the first time. PMID- 12579840 TI - [Solubility behavior of four diastereomeric salts and two amino acids in near critical CO2]. AB - AIM: To establish a suitable condition for extraction of phenylalanine (Phe), 5 hydroxytryptophan (5-OH-Trp) and four diastereomeric salts, (1R,2S)-ephedrine- (2S,3S)-tartaric acid, (1R,2S)-ephedrine-(2R,3R)-tartaric acid, (1S,2S) pseudoephedrine-(2S,3S)-tartaric acid, (1S,2S)-pseudoephedrine- (2R,3R)-tartaric acid in supercritical fluid extraction and to assess the solubilities of Phe, 5 OH-Trp and the four diastereomeric salts in CO2. METHODS: Single-pass method and HPCE. RESULTS: The solubilities of Phe, 5-OH-Trp and the four diastereomeric salts in CO2 were determined over temperature and pressure ranges of 25-50 degrees C and 6.32-34.03 MPa respectively. The experimental results showed that the solubilities of Phe, 5-OH-Trp and the four diastereomeric salts do not increase with density of CO2. There existed a maximum in the critical region of CO2. CONCLUSION: The dramatically high solubilities in the pressure of 6.32-7.78 MPa show a critical behavior, which can be explained by critical characteristic through thermodynamics analysis. The results suggest that the separation of Phe, 5-OH-Trp and the four diastereomeric salts is more efficient in critical region of CO2. PMID- 12579841 TI - [Determination of cyclovirobuxine D by RP-HPLC with precolumn fluorescence derivatization]. AB - AIM: To establish a RP-HPLC method for determination of cyclovirobuxine D. METHODS: Cyclovirobuxine D reacted with a derivative reagent 1-naphthyl isocyanate in chloroform to form fluorescence derivatives, stopped the reaction by adding the mobile phase and then directly injected the solution into the chromatograph to seperate it by RP-HPLC. The analysis was carried out on C18 column, the mobile phase is methanol-water (85:15), the excitation wavelength was set at 305 nm, emission at wavelength 385 nm, and the flow rate was 1 mL.min-1. The effect of several factors including the reaction medium, temperature, time and amount of 1-naphthyl isocyanate on the yield of the derivatization was also investigated systematically. RESULTS: A simple and rapid RP-HPLC method for the simultaneous isolation and analysis of cyclovirobuxine D and its related substances was developed, and the absence of interference between the derivative peak responses of cyclovirobuxine D and its related substances were verified by UV diode array detecter and MS. The linearity was obtained from 0.75 microgram.mL 1 to 2.5 micrograms.mL-1 of cyclovirobuxine D derivatives with a correlation coefficient of 0.9991. The detection limit of cyclovirobuxine D derivative was 1 ng.mL-1, the repeatability of derivatization was good with relative standard derivation no more than 1.2% and derivative was stable within 48 h. The method described conforms to the validation of China Pharmacopiea compendial methods used for pharmaceutical products in general. CONCLUSION: The established method is proved to be reliable quantitative method for the quality control of cyclovirobuxine D. PMID- 12579842 TI - [Determination of wogonin in rat plasma by liquid chromatography-tandem mass spectrometry]. AB - AIM: To develop a sensitive and rapid LC/MS/MS method for the determination of wogonin, an active flavonoid shown to have an inhibitory effect on the proliferation of a carcinoma cell line, in rat plasma after an oral administration. METHODS: Wogonin and daidzein (internal standard) were extracted from plasma directly with n-hexane-diethyl ether (1:4). After liquid-liquid extraction, the analytes of interest were separated on a Diamonsil C18 column. The mobile phase was acetonitrile-water-formic acid (80:20:1) with a flow rate of 0.8 mL.min-1. A Finnigan TSQ (triple stage quadruple) tandem mass spectrometer equipped with an atmospheric pressure chemical ionization (APCI) source was used as detector and was operated in positive ion mode. Selected reaction monitoring (SRM) was used and transitions selected for quantitation were: m/z 284.8-->269.5 for wogonin and m/z 254.7-->198.5 for daidzein. The mass spectrometric conditions were as follows: The temperatures of the vaporizer and heated capillary were 450 degrees C and 250 degrees C, respectively. The corona discharge current was 4.00 microA. Nitrogen was used as the sheath and auxiliary gas, whose settings were 0.6 MPa and 3 mL.min-1, respectively. Argon was used as the collision gas at a pressure of 1.4 Pa. The collision energy of 35 V was chosen for both wogonin and daidzein. RESULTS: The calibration curve was linear over the concentration range of 0.25 to 20.0 ng.mL-1. The limit of quantitation was 0.25 ng.mL-1. Within-day and between-day precision expressed by relative standard deviation (RSD) was 2.2% to 13.1% and 5.9% to 7.3%, respectively, and the accuracy expressed by RE was 0.3% to 1.3%. CONCLUSION: This method proved to be specific, accurate and sensitive enough to be applied to the pharmacokinetic studies of wogonin in rats after a single dose of 5 mg.kg-1 by oral administration. PMID- 12579843 TI - [Pharmacokinetics of crocetin in rats]. AB - AIM: To develop an HPLC method for the determination of crocetin in rat plasma and study the pharmacokinetics in rats. METHODS: Hypersil C18 column (5 microns, 4.6 mm x 200 mm) was used at column temperature 30 degrees C. The mobile phase consisted of methanol-water-acetic acid (75:24.5:0.5) at the flow rate of 1.0 mL.min-1. The UV detection wave length was 423 nm. RESULTS: The calibration curve was linear (gamma = 0.9996) in the range from 0.49 microgram.mL-1 to 7.87 micrograms.mL-1 for crocetin. The mean recovery was 105.2%. The lowest detectable concentration of crocetin was 0.14 microgram.mL-1 (S/N = 3). The RSDs of within day and between-day were all less than 5%. The plasma crocetin was steady. The HPLC method of determination of crocetin in the plasma was established. After single dose of 50 mg.kg-1 ig in 10 rats, the main pharmacokinetic parameters were estimated as follows: T1/2 alpha (30 +/- 6) min, Tmax(65 +/- 16) min, Cmax(5.0 +/ 1.0) microgram.mL-1, AUC0-T(845 +/- 109) microgram.min.mL-1, Vd(5.0 +/- 0.8) L.kg-1. Crocetin was shown to be absorbed into the blood through the gastrointestinal tract. CONCLUSION: This method is quick, precise and reliable. Crocetin was shown to be quickly absorbed in rats. PMID- 12579844 TI - [Interaction between insulin with liposome]. AB - AIM: To study the characteristics of the interaction between insulin and liposome. METHODS: The interaction between insulin and liposome was studied by fluorescence spectra and microcalorimetry methods. The sample of insulin liposome interaction after separation by supper-centrifugalization or gel filtration was determined by fluorescence and HPLC. RESULTS: The results indicate that there was only little increase in fluorescence intensity and no blue shift of peak in fluorescence spectrum. Fluorescence quenching experiments with NaI and acrylamide as quenchers showed that the KSVs (the slope of Strm-Volmer equation) of insulin were more similar to that with added liposome, indicating low interaction between insulin with liposome. The microcalorimetric results indicate that the heat released during the mixture of liposome with insulin, was 1.98 kcal.mol-1, suggesting that the reaction belongs to weak reaction. The quantity of insulin in the insulin-liposome mixture sample after separation by ultracentrifuge or by Sephadex G-75 determined by HPLC, the combination percent was only 0.2%, indicating low interaction between insulin and liposome. CONCLUSION: The interaction between insulin and liposome was weak. PMID- 12579845 TI - [Study on preparation and oral efficacy of insulin-loaded poly(lactic-co-glycolic acid) nanoparticles]. AB - AIM: To investigate the possibility of poly(lactic-co-glycolic acid) as a carrier for the delivery of macromolecular. METHODS: Insulin-loaded poly (lactic-co glycolic acid) nanoparticles (INS-PLGA-NPs) was prepared by a double-emulsion solvent evaporation method. The size distribution was examined by photo correlation spectrometry. The entrapment efficiency was determined by HPLC and important factors that affected the entrapment efficiency were investigated. The loading mechanism of different size nanoparticles was assayed by radioimmunoassay (RIA). INS-PLGA-NPs release behavior in vitro was carried out under sink condition. After oral administration of the nanoparticles to alloxan-induced diabetic rats, its glucose level was determined by glucose oxidize method and the oral pharmacological bioavailability in contrast to s.c. of insulin solution was calculated according to the area over the curve. RESULTS: The INS-PLGA-NPs was prepared with poloxamer 188 as a emulsifier, the mean diameter was 149.6 nm and the polydispersity index was decreased to 0.09. While the entrapment efficiency was increased to 42.8%. Most of the insulin loaded was adsorbed on the surface of the nanoparticles. The release behavior in vitro showed an initial burst effect followed by a slower rate stage. After oral administration of 10 u.kg-1 INS-PLGA NPs, the plasma glucose level decreased significantly after 4 h (P < 0.05), 10 h later the glucose level decreased to the lowest (52.4% +/- 10.2%, P < 0.01) and the relative pharmacological bioavailability is (10.3 +/- 0.8)%. CONCLUSION: PLGA NPs might be used as a new oral carrier for protein drug delivery systems in the future. PMID- 12579846 TI - [Pulmonary delivery of insulin lipid suspension]. AB - AIM: To investigate the relative bioavailability of pulmonary-delivered insulin lipid suspension (INS-LIP-SP) in normal Wistar rats. METHODS: INS-LIP-SP were prepared by two different methods and then delivered to the rat lung using an intratracheal instillation method. Blood glucose levels and INS concentrations in serum were determined by glucose oxidase method and radioimmunoassay method, respectively. The relative pharmacological bioavailability (f%) and relative bioavailability (F%) of INS-LIP-SP were calculated from the area above the curve (AAC) and the area under the curve (AUC) compared with subcutaneous injection of INS solution. RESULTS: The mean particle diameter, span of dispersity and entrapment efficiency of INS-LIP-SP prepared by a membrane-formed with sonication method and a reversed phase evaporation method were 1.91 microns, 0.94 and 16.45% and 2.08 microns, 1.28 and 39.51%, respectively. The values of f% and F% of both INS-LIP-SP were up to 37% and 32%, separately, and there was a statistically significant difference between INS-LIP-SP and INS solution. However, there was no significant difference between the two INS-LIP-SP and the physical mixture of INS solution and blank liposomes. CONCLUSION: The results showed that INS-LIP-SP could achieve higher bioavailability following pulmonary delivery to rats. PMID- 12579847 TI - [A study on genetic variation between wild and cultivated populations of Paeonia lactiflora Pall]. AB - AIM: To discuss the genetic differentiation between wild and cultivated populations of Paeonia lactiflora Pall., and to find the reason for forming the genuineness of Radix Paeoniae rubra (Chishao) and Radix Paeoniae alba (Baishao). METHODS: Forty three representative samples of P. lactiflora from 11 localities were analyzed by RAPD method with 21 random primers. According to RAPD results, the genetic diversity and genetic differentiation of P. lactiflora were detected by the percentage of polymorphic sites (PPB), the coefficient of gene differentiation (GST) and analysis of molecular variance (AMOVA). RESULTS: (1) At species level, the PPB of P. lactiflora was 85.26%, the Nei's gene diversity (Ht) was 0.166. The PPB in wild population (WP) was 77.61%, which was more than that (54.96%) in cultivated population for medicine (MP), and that (61.76%) in cultivated population for ornament (OP). (2) AMOVA showed that 29.50% of the genetic diversity resulted from differentiation among populations. Pairwise Phist distance (0.3632) between WP and MP was furthest, while that (0.0973) between MP and OP was closest, indicating population differentiation was significant (P < 0.001). (3) In general, cluster analysis revealed that the samples of P. lactiflora were divided in wild and cultivated groups (except for 39). In WP, individuals of Duolun were separated from those of other localities. In MP, the clusters of samples corresponded well with their own habitats. CONCLUSION: In addition to environmental factor, genetic differentiation should be the main cause for the genuineness of "Chishao" and "Baishao". Because of over collection and worse habitat, P. lactiflora in Duolun, whose root is the famous Chinese Geo herbal--"Duolun Chishao", is progresively rare. So, it should become the endangered germplasm resource to protect. PMID- 12579848 TI - [Application of LC-MS in the analysis of constituents of crude plant extracts]. PMID- 12579849 TI - [Problems and solutions in pegylation of protein and peptide drugs]. PMID- 12579850 TI - [Effect of ginsenoside Rg1 on learning and memory impairment induced by beta amyloid peptide(25-35) and its mechanism of action]. AB - AIM: To study the effect of ginsenoside Rg1 on the learning and memory impairment in mice induced by aggregated beta-AP(25-35). METHODS: Mice were administered Rg1(5, 10 mg.kg-1, i.p.) for 10 d and control mice received daily i.p. injections of saline after the intracerebroventricular injection of aggregated beta-AP(25 35). After the final treatment, passive avoidance and performance in the Morris water maze (MWM) were assessed. and the activity of cortical and hippocampal ChAT and AchE were detected after the final behavior test. RESULTS: Ginsenoside Rg1 (5, 10 mg.kg-1, i.p.) significantly ameliorated the learning and memory impairment induced by beta-AP(25-35). Rg1 (5, 10 mg.kg-1) decreased the latencies and swim distances of mice to reach a hidden platform and improved the corresponding changes in search strategies occurred in the Morris water maze, and Rg1 (10 mg.kg-1, i.p.), increased step-through latencies also. Biochemical analysis showed that Rg1 (5, 10 mg.kg-1, i.p.), prevented the cortical and hippocampal ChAT activity decline induced by beta-AP(25-35), and showed inhibition of the activity of AchE, although beta-AP(25-35) showed no effect on the cortical and hippocampal AchE activity. CONCLUSION: These data showed that ginsenoside Rg1 significantly improved the learning and memory impairment induced by beta-AP(25-35), and this effect could be attributed to its inhibition of AchE and increase of ChAT activity. PMID- 12579851 TI - [Effect of melatonin on release of beta-endorphin, norepinephrine and 5 hydroxytryptamine in rat brain]. AB - AIM: In order to explore the mechanism of action of melatonin to induce analgesia, the present study was undertaken to observe the effects of melatonin on the release of beta-endorphin (beta-Ep), norepinephrine (NE) and 5 hydroxytryptamine in rat brain. METHODS: With the measurement of pain threshold, push-pull perfusion technique and radioimmunoassay were used to determine the immunoreactive beta-Ep content in the perfusate from the third ventricle of rat brain. The contents of 3-methoxy-4-hydroxyphenyl glycol (MHPG) and 5 hydroxyindole acetic acid in the microdialysate from rat brain were measured by techniques of in vivo microdialysis and high performance liquid chromatography with electrochemical detection. RESULTS: The immunoreactive beta-Ep content in the perfusate from the third ventricle of rat brain was increased significantly (P < 0.05) following an intraperitoneal administration of 110 mg.kg-1 of melatonin, with the increase of pain threshold. The MHPG and 5-HIAA contents in the microdialysate from the periaqueductal gray (PAG) or the hypothalamus were not changed after the administration of melatonin. CONCLUSION: Melatonin may promote the release of beta-Ep in brain, which may be one of the mechanisms of the analgesic action of melatonin. The analgesic action of melatonin may not be related to the release of NE and 5-HT in the PAG or the hypothalamus. PMID- 12579852 TI - [Inhibition of Na+/Ca2+ exchange by tetrapeptide FMRFa in intact rat ventricular myocytes]. AB - AIM: To study the effects of Phe-Met-Arg-Phe-NH2(FMRFa) on Na+/Ca2+ exchange and its specificity for Na+/Ca2+ exchange in rat ventricular myocytes. METHODS: Na+/Ca2+ exchange current and other currents of ion channels were measured using whole cell voltage clamp techniques. RESULTS: A dose-related inhibition of tetrapeptide FMRFa on Na+/Ca2+ exchange was observed in rat ventricular myocytes. Inward and outward INa+/Ca2+ were inhibited by 60.1% and 56.5%, respectively, at highest concentration (100 mumol.L-1) and its IC50 were 20 mumol.L-1 and 34 mumol.L-1 in inward and outward INa+/Ca2+, respectively. Inward and outward INa+/Ca2+ were inhibited 38.7% and 34.9%, respectively, at FMRFa 5 mumol.L-1. FMRFa 5 mumol.L-1 and 20 mumol.L-1 did not affect L-type calcium current, sodium current, transient outward current and inward rectifier potassium current. CONCLUSION: These data indicate that FMRFa is a specific inhibitor of Na+/Ca2+ exchange in intact rat ventricular myocytes. PMID- 12579853 TI - [Inhibitory effect of boanmycin on the growth of colon carcinoma 26 and hepatic metastasis in mice]. AB - AIM: To investigate the therapeutic effect of boanmycin (BAM, bleomycin A6) on colon carcinoma 26 and its hepatic metastasis in mice. METHODS: A series of models including subcutaneous transplantation, orthotopic transplantation in cecum subserosa, intra-hepatic transplantation of tumor, and intrasplenic transplantation of tumor accompanied with hepatic metastases were employed. In addition, LEICA Q 500IW image analysis system was used to determine the area of metastatic lesions in the liver in histopathological sections. RESULTS: BAM at 5 mg.kg-1 and 2.5 mg.kg-1 inhibited the growth of subcutaneous tumors by 78.7% and 61.9%, and the orthotopic tumors by 99.4% and 90.0%; and the intra-hepatic tumors by 86.9% and 75.7%, respectively. Determined by the numbers of metastatic nodules, BAM at 10 mg.kg-1 and 5 mg.kg-1 inhibited hepatic metastases from intra splenic transplantation by 97.6% and 56.8%; and evaluated by image analysis of metastatic lesions, by 100% and 63.3%, respectively. CONCLUSION: Boanmycin is highly effective in a panel of models including the subcutaneous, orthotopic, intra-hepatic transplanted tumors, and hepatic metastases of murine colon carcinoma 26. PMID- 12579854 TI - [Inhibitory effects of D-polymannuronic sulfate on proliferation of rat vascular smooth muscle cells and its related mechanisms of action in vitro]. AB - AIM: To investigate the inhibitory effects of D-polymannuronic sulfate (DPS) on the proliferation of rat vascular smooth muscle cells (VSMC) induced by basic fibroblast growth factor (bFGF) or interleukin-1 (IL-1) and its related mechanisms. METHODS: Rat aortic smooth muscle cells pretreated with DPS in concentrations ranging from 0.001 microgram.mL-1 up to 100 micrograms.mL-1 were incubated at 37 degrees C for 24 h, followed by addition of bFGF (50 ng.mL-1) or IL-1 (50 U.mL-1) for another 24 h. The effects of DPS on the proliferation of VSMC were evaluated by MTT assays. VSMC were pretreated with DPS in concentrations ranging from 0.001 microgram.mL-1 up to 1 microgram.mL-1, followed by addition of L-NAME (0.1 microgram.mL-1) or bFGF (50 ng.mL-1) for 24 h. Supernatant nitric oxide (NO) was determined with NO assay kit, while supernatant angiotensin II (Ang II) and endothelin-1 (ET-1) were measured by radioimmunoassay. RESULTS: DPS exerted antiproliferative effects at concentrations ranging from 0.01 microgram.mL-1 to 10 micrograms.mL-1, and its maximal effect was observed at the concentration of 1 microgram.mL-1. Also, the suppressing actions of DPS on the proliferation of VSMC were diminished by increasing the concentrations of bFGF or IL-1. Furthermore, DPS increased NO synthesis and decreased Ang II and ET-1 contents released from VSMC in a concentration-dependent manner. CONCLUSION: DPS afforded the antiproliferative effects on bFGF- or IL-1-treated VSMC and its underlying mechanisms were associated with enhancement of NO synthesis and decrement of Ang II and ET-1 production/release in vitro. PMID- 12579855 TI - [Effect of KB-R7943 on Na(+)-Ca2+ exchange current in guinea pig ventricular myocytes]. AB - AIM: To study whether KB-R7943 has selective inhibitory effect on the inward and outward Na(+)-Ca2+ exchange current (INa-Ca) in guinea pig ventricular myocytes. METHODS: Through setting up the model of intracellular Na(+)-overload during myocardial ischemia and reperfusion, the current-voltage relationship of INa-Ca was recorded using whole-cell patch clamp technique under bi-directional ionic conditions. RESULTS: Currents were elicited by a declining ramp pulse depolarized immediately from holding potential of -40 mV to +60 mV, then repolarized to -100 mV at a speed of 80 mV.s-1 and returned to the holding potential under bi directional ionic conditions, while the [Na+] was 25 mmol.L-1 in the pipette solution. The currents increased time-dependently and voltage-dependently which reached from (2.51 +/- 0.15) pA.pF-1 to (5.94 +/- 0.13) pA.pF-1 at mV and from ( 1.92 +/- 0.13) pA.pF-1 to (-3.17 +/- 0.16) pA.pF-1 at -80 mV (n = 12) after 3 min and there is no significant run-down of the current. KB-R7943 10(-6) mol.L-1 was found to decrease the current to (4.62 +/- 0.05) pA.pF-1 by 29.4% at mV and to ( 2.30 +/- 0.18) pA.pF-1 by 22.1% at -80 mV (n = 5) after 5 min. While 10(-5) mol.L 1 KB-R7943 was shown to decrease the current to (3.13 +/- 0.03) pA.pF-1 by 61.7% at mV and to (-1.62 +/- 0.03) pA.pF-1 by 56.9% at -80 mV (n = 7). CONCLUSION: KB R7943 can block INa-Ca in guinea pig ventricular myocytes. But, it did not show selective inhibition effect on inward and outward currents. PMID- 12579856 TI - [Synthesis and anticancer activity of 3-[(3'-methyl-4'-(substituted phenyl)-1',3' butadienyl] indole derivatives]. AB - AIM: To study the synthesis and anticancer activity of 3-[(3'-methyl-4' (substituted phenyl)-1',3'-butadienyl] indole derivatives. METHODS: Electrophilic substitution, aldol-condensation, selective-reduction, phase-transfer Wittig reaction and hydrolysis reaction were used in the synthesis of the title compounds. RESULTS: Eleven compounds of 3-[(3'-methyl-4'-(substituted phenyl) 1',3'-butadienyl] indole were synthesized. They are new compounds. Compound 8 showed different inhibitory effects on HL-60, HCT-8 and Bel7402 cell lines in vitro, and its inhibitory rate of antiinflammation was 100% at 10(-5) mol.L-1 concentration. CONCLUSION: Compound 8 showed high anticancer and antiinflammatory activities, and is worth further studying. PMID- 12579857 TI - [Studies on the chemical constituents of Eclipta prostrata (L)]. AB - AIM: To study the chemical constituents of Eclipta prostrata (L). METHODS: The constituents of E. prostrata were systematically separated with the Bohlmann method and percolation and hot extraction methods, and various chromatographies. The structures were elucidated by chemical and spectroscopic means. RESULTS: Ten compounds were isolated from the Eclipta prostrata. Their structures were determined as wedelolactone (1), demethylwedelolactone (2), isodemethylwedelolactone (3), alpha-formylterthienyl (4), strychnolactone (5), beta-sitosterol (6), nonacosanol (7), stearic acid (8), lacceroic acid (9), 3,4 dihydoxy benzoic acid (10). Fourteen ocmpounds, including hydrocarbons and its esters were identified by GC-MS from the least polar fractions. CONCLUSION: Compound 3 is a new coumestan named isodemethylwedelolactone. Compounds 2-10 and compounds characterized by GC-MS analysis were obtained for the first time from Eclipta prostrata. PMID- 12579858 TI - [Chemical constituents from the root of Rubus chroosepalus]. AB - AIM: To investigate the chemical constituents of Rubus chroosepalus Focke. METHODS: The methanol extracts were suspended in H2O and then extracted with EtOAc. Column chromatography was used for separation and purification, while spectral analysis was used for identification. RESULTS: Seven compounds were isolated and identified as 2 alpha, 3 beta-dihydroxy-urs-12, 19-dien-23, 28-oic acid (I), 2 alpha, 3 beta, 23-trihydroxy-urs-12,18-dien-28-oic acid (IIa), 2 alpha, 3 beta, 23-trihydroxy-urs-12,19-dien-28-oic acid (IIb), 2 alpha, 3 alpha dihydroxy-urs-12,18-dien-28-oic acid (IIIa), 2 alpha, 3 alpha-dihydroxy-urs-12,19 dien-28-oic acid (IIIb), and the acetonide of IIIa and IIIb (IVa and IVb). CONCLUSION: I was found to be a new pentacyclic triterpenoid acid. PMID- 12579859 TI - [Isolation and identification of steroidal saponins from Dioscorea panthaica Prain et Burkill]. AB - AIM: To investigate the chemical constituents of the rhizomes of Dioscorea panthaica Prain et Burkill and look for new active compounds. METHODS: The compounds were isolated with silica gel and HPLC, and their structures were elucidated on the basis of chemical evidences and spectral analysis (IR, ESI-MS, 1HNMR, 13CNMR, DEPT, 1H-1H COSY, HMQC, HMBC, NOESY). RESULTS: Three steroidal saponins were isolated from the rhizomes of Dioscorea panthaica. They were elucidated as 26-O-beta-D-glucopyranosyl-3 beta, 26-diol-23(S)-methoxy-25(R) delta 5,20(22)-diene-furosta-3-O-[alpha-L-rhamnopyranosyl-(1-->2)- O-alpha-L rhamnopyranosyl-(1-->4)]-beta-D-glucopyranoside (I), pseudoprotodioscin (II), 26 O-beta-D-glucopyranosyl-3 beta, 26-diol-25 (R)-delta 5,20(22)-diene-furosta-3-O alpha-L-rhamnopyranosyl- (1-->2)-O-beta-D-glucopyranoside (III). CONCLUSION: Compound I is a new steroidal saponin and was named as dioscoreside C. Compound II and III were obtained from this genus for the first time. PMID- 12579860 TI - [High performance liquid chromatography/electrospray ionization mass spectrometric characterization of recombinant L-asparaginase II]. AB - AIM: To characterize the primary structure of recombinant L-asparaginase II product. METHODS: The molecular weight of the protein was measured by pneumatically-assisted electrospray ionization mass spectrometry with flow injection mode. Subsequently, tryptic peptide mapping was performed by high performance liquid chromatography on a C8 column with tandem UV and MS detection. An easy-to-use and simple denaturation process with trichloroacetic acid was conducted prior to tryptic digest so as to release the digest resistance from the protein structure. The amino acid sequences of the tryptic peptides were elucidated based on their in-source collision-induced dissociation spectra. RESULTS: The measured molecular mass was different from the theoretical value. Three amino acid variations were unambiguously detected along the peptide backbone derived from the gene-encoding sequence. CONCLUSION: This paper revealed that LC/ESI/MS had provided a promising and robust technique in primary structure analysis and quality control of DNA-derived recombinant protein pharmaceuticals. PMID- 12579861 TI - [Determination of amlodipine in human plasma by liquid chromatography-tandem mass spectrometry]. AB - AIM: To develop a sensitive and specific LC/MS/MS method for determination of amlodipine in human plasma. METHODS: Amlodipine and internal standard 4' hydroxypropafenone were extracted from plasma using liquid-liquid extraction, then separated on a Zorbax C8 column. The mobile phase consisted of acetonitrile water-formic acid (75:35:1), at a flow-rate of 0.4 mL.min-1. A Finnigan TSQ tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode. Selected reaction monitoring (SRM) using the precursor-->product ion combinations of m/z 409-->238 and m/z 358 ->116 was used to quantify amlodipine and internal standard, respectively. RESULTS: The linear calibration curves were obtained in the concentration range of 0.4-16.0 ng.mL-1. The limit of quantification was 0.4 ng.mL-1. Each plasma sample was chromatographed within 3.7 min. The method was successfully used in several pharmacokinetic studies for amlodipine. More than 1,500 plasma samples were assayed within two weeks. CONCLUSION: The method is proved to be suitable for clinical investigation of amlodipine pharmacokinetics, which offers advantages of specificity, speed, and greater sensitivity over the previously reported methods. PMID- 12579862 TI - [SFE-CGC determination of tanshinone in Salvia miltiorrhiza Bunge]. AB - AIM: To analyze tanshinone IIA in Salvia miltiorrhiza Bunge by supercritical fluid extraction (SFE) coupled with capillary gas chromatography (CGC). METHODS: The solution pressure was predicted by solubility parameter theory, the extraction conditions were confirmed by orthogonal design in which temperature, volume of modifier and dynamic extracting volume had been studied in three levels. RESULTS: This method is quick, convenient, precise and accurate, added sample recovery is 95.3%, RSD = 4.27%. CONCLUSION: Compared with the method of ultrasonic extraction, significantly better qualitative and quantitative result could be obtained by SFE-CGC and expend less organic solvent then make less pollution. PMID- 12579864 TI - [Study on the release mechanism of fenoprofen calcium from hydrophillic sustained release matrix]. AB - AIM: To study the release mechanism of fenoprofen calcium (FC) from hydroxypropylmethylcellulose (HPMC) matrices. METHODS: The release of FC and the erosion properties of hydrophillic matrices containing HPMC was examined at different paddle speed. The release mechanism of FC was further confirmed by evaluating the n value in Peppas equation. RESULTS: The results indicate that the release of FC and the erosion of matrices exhibit zero order kinetic equation, and it exhibits line relationship between them. CONCLUSION: In the first 40 min, FC mainly released by diffusion and erosion from HPMC matrix, while it was controlled by the rate of tablet erosion after 50 min. PMID- 12579863 TI - [Application of triangular phase diagrams to investigate the formulation conditions of pharmaceutical microemulsions]. AB - AIM: To establish innovational triangular phase diagrams for preparation of pharmaceutical microemulsion (ME) with low toxicity as a result of reducing the amount of the emulsifiers. METHODS: Lecithin and polyoxyethylene octyl phenyl ether (OP) were used as emulsifiers (E), absolute alcohol and n-octyl alcohol as assistant emulsifiers (A), ethyl oleate and olive oil as the oil phase (O). For the innovational triangular phase diagrams, lecithin-ethyl oleate-water and absolute alcohol (optimized ratio W/A = 0.6) system, and OP(olive oil and n-octyl alcohol (O/A = 0.34)-water system were used for preparation of the ME. While for the traditional triangular phase diagrams, OP and n-octyl alcohol (E/A = Km = 1.5)-olive oil-water system, lecithin and absolute alcohol (E/A = Km = 1.12) ethyl oleate-water system were used. Then, the corresponding amounts of the emulsifiers used in both methods were compared. RESULTS: In the ME of lecithin systems by the innovational and the traditional methods, the amounts of emulsifier are 6%-28% and 28%-29.6%, respectively. In the ME of OP systems by the innovational and the traditional methods, the amounts of emulsifier are 23%-40% and 27.6%-49.8%, respectively. CONCLUSION: The method of innovational triangular phase diagram for preparation of ME is superior to the traditional method in finding out the lower amounts of emulsifiers for ME, and is a feasible method with guiding significance for preparing ME. PMID- 12579865 TI - [A comparison of the its sequences of the Tibetan medicine "zang yin chen"- Swertia mussotti and its adulterant species]. AB - AIM: To identify the medicine and search for new medicine resource at the molecular level, the ITS of "Zang Yin Chen"--Swertia mussotti and its adulterant species were sequenced. METHODS: The double-stranded DNA was amplified using PCR systems 9,600 kits and sequenced on an ABI 377 automated sequencer from both directions. RESULTS: The ITS sequences of S. mussotti in different populations showed no variation. It has the unique ITS sequence and shows distinct difference from its adulterant species. In the phylogenetic tree based on the ITS data of S. mussotti and all vicarious species constructed by Paup, S. franchetiana and S. mussotti clustered together with high bootstrap support. CONCLUSION: ITS sequences can be used for the molecular authentication between the S. mussotti and its adulterant species. S. franchetiana can be regarded as a new medicine resource of "Zhang Yin Chen". PMID- 12579866 TI - [Pharmacokinetics of meloxicam in healthy Chinese volunteers]. AB - AIM: To assess the pharmacokinetic profile of single doses of meloxicam in healthy Chinese volunteers. METHODS: The plasma concentrations of meloxicam after an oral dose of 15 mg to twenty healthy male volunteers were analyzed by means of a validated HPLC method. The pharmacokinetic parameters were subjected to Shapiro Wilk test to determine whether these data were fitted to a normal distribution. RESULTS: The twenty volunteers can be classified into extensive metabolizers and poor metabolizers according to pharmacokinetic parameters. The main parameters in the two groups obtained were as follows: T 1/2 were 21 +/- 4 and 38 +/- 9 h, AUC0 infinity were 49 +/- 10 and 110 +/- 8 micrograms.h.mL-1, respectively. Even the AUC data in extensive metabolizers were 1.7 times as that reported in White volunteers following the same doses of meloxicam. CONCLUSION: There were significant individual differences in the pharmacokinetics of meloxicam in Chinese volunteers, which may be due to the genetic polymorphism of CYP2C9. PMID- 12579867 TI - [Research development of the interaction of small molecules with nucleic acids]. PMID- 12579868 TI - [Effect of stilbene polymer (Gn-3) on experimental liver injuries in mice]. AB - AIM: To study the protective effect of Gn-3 (a stilbene polymer isolated from Gnetum parvifolium) against liver injury induced by CCl4, N-acetyl-p-aminophenol (APAP) and Bacillus Calmette-Guerin (BCG) plus bacterial lipopolysaccharide (LPS) in mice. METHODS: The experimental model of liver injury were induced by 0.1% CCl4 i.p. (10 mL.kg-1.d-1 for 3d), APAP i.p. (150 mg.kg-1) or BCG (5 mg) plus LPS (7.5 micrograms) in mice. The levels of ALT in serum, MDA and GSH in liver tissues were detected. The histopathologic changes were observed by light microscope. RESULTS: Gn-3 was shown to markedly reduce the elevated serum ALT levels, liver tissue MDA and improve the histopathological changes in all the three experimental liver injury models. No effect of Gn-3 was observed on the liver GSH level in liver injury mice. CONCLUSION: Gn-3 was found to inhibit the development of liver injury caused by CCl4, APAP, or BCG plus LPS. This means that Gn-3 has liver protective effects. PMID- 12579869 TI - [Cardiac-hemodynamic effects of M3 receptor agonist on rat and rabbit hearts]. AB - AIM: To study the activation of choline on M3-R in heart and observe the hemodynamic changes of rat and rabbit. METHODS: A cardiac catheter was inserted into the left ventricular cavity via the right carotid artery, then the HR, LVSP, LVEDP, and +/- dp/dt were measured using a polygraph system. RESULTS: Choline was shown to decrease the hemodynamic assessments, such as HR, + dp/dt, LVSP and LVEDP. while the M3-R antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine methiodide) showed little effect on these assessments. It was found to reverse the hemodynamic effects of choline. CONCLUSION: M3 receptor agonist can produce negative inotropic and chronotropic effects on the heart of rat and rabbit. PMID- 12579870 TI - [Allele specific amplification for CYP2D6 gene related to intermediate metabolizer in Chinese subjects]. AB - AIM: To establish an allele specific PCR amplification (ASA-PCR) for determination of the genotype of CYP2D6* 10B polymorphism in Chinese subjects. METHODS: CYP2D6* 10B alleles of 65 healthy Chinese subjects were analyzed by a two-step PCR assay and the correlation of genotype and phenotype was studied. RESULTS: There were 20 CYP2D6* 10B heterozygous genotypes subjects (wt/m) in 35 very extensive metabolizers (VEMs), which consisted the major part of VEM subjects (57%). Meanwhile, 20 subjects consisting 69% of 29 intermediate metabolizers were CYP2D6* 10B homozygous mutant genotypes (m/m). The poor metabolizer was also m/m. The metabolic ratio of CYP2D6* 10B m/m subjects were larger than wt/m and wild type, the values were -1.49 +/- 0.54, -2.20 +/- 0.49 and -2.47 +/- 0.61 (P < 0.01). CONCLUSION: PCR-ASA was shown to be a rapid and specific method. It can be used to study the genetic polymorphism, especially CYP2D6 intermediate metabolism. PMID- 12579871 TI - [Effects of ginkgolide B on arachidonic acid metabolizing enzymes and level of intracellular calcium in rat polymorphonuclear leukocytes]. AB - AIM: To investigate the effects of ginkgolide B on arachidonic acid (AA) metabolizing enzymes and the level of intracellular calcium in rat polymorphonuclear leukocytes. METHODS: Intracellular free calcium was quantitated by Fura-2 fluorescence technique. Phospholipase A2 (PLA2) activity was determined by incorporating 3H-arachidonic acid in leukocytes. 5-Lipoxygenase (5-LO) activity was evaluated by RP-HPLC. RESULTS: In comparison with control, ginkgolide B at final concentration of 0.1-10 mumol.L-1 inhibited A23187 induced AA release by 10.9%-22.2%; at final concentration of 0.1-50 mumol.L-1, ginkgolide B inhibited LTB4 and 5-HETE synthesis stimulated by PAF by 29.4%-88.8% and 26.2% 89.3% respectively. At the final concentration of 0.1-100 mumol.L-1, ginkgolide B decreased the rise of intracellular calcium level induced by pletelet activating factor (PAF) and N-formyl-methionine-leucine-phenglalanine (fMLP) by 13.9%-51.4% and 2.2%-36.6%, respectively. CONCLUSION: Ginkgolide B was found to significantly inhibit PLA2 and 5-LO activities, as well as the increase of the intracellular calcium induced by PAF. PMID- 12579872 TI - [Poteintation of vincristine-induced apoptosis by tetrandrine, neferine and dauricine in the human mammary MCF-7 multidrug-resistant cells]. AB - AIM: To investigate the poteintation of vincristine-induecd apoptosis by tetrandrine, neferine and dauricine isolated from Chinese medicinal plants in the human mammary MCF-7 multidrug resistant cells. METHODS: The apoptotic cells were detected by fluorescent staining of a combination of Hoechst 33342 and propidium iodide (PI), flow cytometry and agarose electrophoresis. RESULTS: The apoptotic cells induced by vincristine alone accounted for about 10% of all the cancer cells, while the percentage of apoptotic cells induced by a combination of vincristine with tetrandrine, neferine, or dauricine was found to be significantly higher than that by vincristine alone, and their reversal effects were positively correlated with the drug concentration and the exposure time. In addition, tetrandrine was shown to be the most potent in the reversal efficacy among the three compounds to be tested for apoptosis in vitro. CONCLUSION: Tetrandrine, neferine and dauricine showed obvious potenitiation of vincristine induced apoptosis in the human mammary MCF-7 multidrug-resistant cells. PMID- 12579873 TI - [Synthesis and biological activity of 2-[(substituted phenyl)vinyl] indole derivatives]. AB - AIM: To synthesize derivatives of 2-[substituted phenyl)vinyl] indole and find compounds with biological activities by screening in vitro. METHODS: Twenty-one compounds of 2-[(substituted phenyl) vinyl] indole were synthesized by reduction, oxidation and Witting reaction. MS, 1HNMR and elemental analysis were used to determine the structures of the new compounds. RESULTS: These compounds are new ones. CONCLUSION: Six compounds (3, 9, 11, 13, 18 and 20) showed effects on some different receptors, such as alpha 2, D2 and H1, and is worth further studying. PMID- 12579874 TI - [Synthesis and activities of 4-deoxy-4 beta-arylmehtylene sulfonylamido-4' demethylpodophyllotoxins]. AB - AIM: To find new anticancer drug based on the structure of podophyllotoxin. METHODS AND RESULTS: 4-Deoxy-4-beta-arylmethylenesulfonylamido-4'- demethylpodophyllotoxins were (1-15) synthesized and their antitumour activities against KB cells and L1210 leukemia cells were tested. CONCLUSION: These compounds are new compounds. Among them, compounds 2, 3, 8, 9, 11 and 12 showed antitumour activities. When halides exist on the aryl ring of substituted sulfonyl group, the corresponding compound has relatively higher activities. The stereo factor is important for the activities. PMID- 12579875 TI - [Studies on the glycoconjugates and glycans from Lycium barbarum L in inhibiting low density lipoprotein (LDL) peroxidation]. AB - AIM: To determine the effects of glycoconjugates and their glycans from Lycium barbarum L. on inhibiting low density lipoprotein (LDL) peroxidation. METHODS: Using Cu(2+)-induced oxidation as a model, the oxidative production of thiobarbituric acid-reactive substances (TBARS) and the LDL electrophoresis migration on agarose gel were measured. RESULTS: The effects of glycoconjugates and their glycans from Lycium barbarum L. on inhibiting LDL peroxidation were different, among them, glycoconjugate LbGp5 showed the best effect on inhibiting LDL peroxidation. CONCLUSION: The glycoconjugates can inhibit LDL peroxidatin while their glycans showed no effects on inhibiting LDL peroxidation. PMID- 12579876 TI - [Physico-chemical properties and biological activities of a glycoconjugate SPPA-1 from Spirulina platensis]. AB - AIM: To isolate polysaccharide from Spirulina platensis and determine its sugar position, molecular weight and biological activities. METHODS: Sephadex G-75 and CM-Sephadex C-50 were used. The sugar position was analyzed by gas chromatography, the molecular weight was determined by GPC. The homogeneity of this glycoconjugate was determined by HPLC and CE. IR and NMR spectra were used to determine the glycosidic linkage. RESULTS: SPPA-1 was a homogeneous glycoconjugate, its carbohydrate chain was composed of alpha-glucopyranan, carbohydrate content is 91.70%, Nitrogen content was 0.96%, the molecular weight was 69.00 x 10(4), SPPA-1 could eliminate O2-. radicals. CONCLUSION: SPPA-1 was an antioxidative glycoconjugate from Spirulina platensis. PMID- 12579877 TI - [Study of sesquiterpene alkaloids from Tripterygium wilfordii Hook.f]. AB - AIM: To study the chemical composition of Tripterygium wilfordii Hook.f. METHODS: Column chromatography was used to separate the chemical constituents. UV, IR, MS, HRMS, 1HNMR, 13CNMR (COM and OFR), 1H-1H COSY, 1H-13C COSY, 2D-NOESY and 1H-13C COLOC were used to determine the structures of the isolated constituents. RESULTS: Two sesquiterpene alkaloids were isolated and their structures were elucidated as euonine and wilfordconine on the basis of spectral evidence. CONCLUSION: Wilfordconine, a new sesquiterpene alkaloid, was shown to be immunosuppressive. PMID- 12579878 TI - [Studies on the chemical constituents of Uncaria yunanensis Hsia. C.C]. AB - AIM: To research the chemical constituents from dried roots of Uncaria yunanensis Hsia. C. C. METHODS: Modern chromatography was used to isolate chemical components. Their structure were identified by spectral analysis. RESULTS: Seven compounds were isolated and identified as 3 beta, 6 beta, 19 alpha-trihydroxyurs 12-en-28 oic acid (I), 23-nor-24-esomethylene-3 beta, 6 beta-19 alpha trihydroxyurs-12-en-28 oic acid (II), 3-oxo-6 beta, 19 alpha-dihydroxyurs-12-en 28 oic acid (III), oleanic acid (IV), 5,7,3',4'-tetrahydroxy-flavan-3-ol (V), beta-yohimbine (VI) and diangoutengjian I (VII). CONCLUSION: All of the above compounds were isolated for the first time from the root of this plant. Among them, compound VII is a new one. PMID- 12579879 TI - [Determination of pravastatin in rat liver by RP-HPLC]. AB - AIM: To establish a reverse-phase high performance liquid chromatographic method for the determination of pravastatin in rat liver. METHODS: An aliquot of 5 g liver homogenates, spiked with triamcinolone acetonide (internal standard), was extracted by solid-phase extraction with Bond Elut C18 columns. Chromatography was performed using a C18 reverse-phase column with mobile phase of Na2HPO4 buffer sdution (0.035 mmol.L-1, pH 3.0)-acetonitrile (155:42). RESULTS: The linear equation was Y = 0.1843X-4.238 x 10(-3) (gamma = 0.9934) in the range of 0.05-10 micrograms.g-1 liver. The limit of detection for pravastatin was 13 ng.mL 1 (signal-to-noise ratio of 3), and the limit of quantification for pravastatin in liver homogenate was 50 ng.g-1 liver (RSD < 20%). The average extraction recovery of pravastatin from liver at different concentrations was 80.8%, and the average inter-day precision was 11%. This procedure was applied to assay pravastatin in rat liver which were collected from Lewis rats at different times after administration of pravastatin (ig 20 mg.kg-1). CONCLUSION: The method was sensitive and is feasible for the pharmacokinetic and distribution study of pravastatin. PMID- 12579880 TI - [Electrochemical behavior of ofloxacin at Pt/GC ion implantation modified electrode and its application]. AB - AIM: To study the electrochemical behavior of ofloxacin at Pt/GC ion implantation modified electrode. METHODS: With Pt/GC ion implantation modified electrode as working electrode, the behavior of ofloxacin was studied by voltammetry in 0.40 mol.L-1 KCl solution. RESULTS: A sensitive reductive peak of ofloxacin was obtained by linear sweep voltammetry. The peak potential was -1.35 V (vs SCE). The peak current was proportional to the concentration of ofloxacin over the range of 1.0 x 10(-6)-3.0 x 10(-5) mol.L-1 with the detection limit of 5.0 x 10( 7) mol.L-1. The behavior of reduction wave was studied and applied to determination of ofloxacin in tablets. CONCLUSION: The reduction process was irreversible. The element composition, atomicity form and depth of distribution at the surface of Pt/GC electrode were determined by Auger electron spectroscopy (AES), X-ray photoelectron spectroscopy (XPS) and scanning electron microscope (SEM). The catalysis behavior and reaction mechanism at Pt/GC modified electrode was also studied. PMID- 12579881 TI - [HPCE determination of trimebutine maleate in rat plasma and its pharmacokinetics]. AB - AIM: To develop a method for the determination of trimebutine maleate in rat plasma by using high performance capillary electrophoresis. The method was employed to pharmacokinetic analysis of trimebutine maleate. METHODS: Plasma samples were deproteinized with acetonitrile (containing ephedrine hydrochloride as internal standard) and the supernatant was dried under N2 stream at 50 degrees C. The residue was dissolved with methanol-water (1:1) and injected into the capillary by siphon. The electrophoresis was performed in uncoated fused-silica capillary and the voltage was 10 kV. The running buffer was 0.03 mol.L-1 NaH2PO4(pH 6.0). The eluate was detected at 214 nm by UV detection. RESULTS: The recovery for trimebutine maleate in rat plasma was 72.8%-87.9%. The calibration curve in plasma was linear over the range 5-200 micrograms.L-1. The limit of quantitation was 5 micrograms.L-1. The intraday relative standard deviation (n = 6) and the interday relative standard deviation (n = 18) were less than 14%. The highest concentration in plasma was observed at 30 min after ig trimebutine maleate to rats. The pharmacokinetic results were AUC0-infinity = 8 micrograms.min.mL-1, T1/2(Ke) = 173 min and Ke = 5.6 x 10(-3) min-1. CONCLUSION: The method is accurate, sensitive and suitable for pharmacokinetic study of trimebutine maleate. PMID- 12579882 TI - [Pharmacokinetics and bioavailability of clinafloxacin in rats]. AB - AIM: To study the pharmacokinetics and bioavailability of clinafloxacin in rats. METHODS: The drug concentration was determined by HPLC. The main pharmacokinetic parameters were obtained by 3P87 program. An RP-C18 was used as the stationary phase. The mobile phase was a mixture of acetonitrile-0.05 mol.L-1 citric acid triethylamine (pH 2.5) (20:80). The flow rate was 1.0 mL.min-1. The UV absorbance detector was set at 300 nm. RESULTS: A good linearity was obtained from 0.03-20 micrograms.mL-1 of clinafloxacin in rat plasma with gamma = 0.9998. The plasma concentration--time curve of clinafloxacin conformed to one compartment open model. After ig administration of 50 mg.kg-1 and 100 mg.kg-1 dose of clinafloxacin in six rats, mean Cmax and AUC values increased in proportion to dose. Mean T1/2 appeared to be independent of dose. Mean AUC was 65 +/- 6 and 27 +/- 4 micrograms.h.mL-1 respectively after i.v. and ig administration of 100 mg.kg-1 dose. The extent of bioavailability (F) of clinafloxacin was 42%. CONCLUSION: The results of the pharmacokinetic study of clinafloxacin showed that it exhibited first order kinetic characteristics and the bioavailability is low. PMID- 12579883 TI - [Analysis of the metabolite of 7-(4-chlorbenzyl)-7,8,13,13a-tetrahydroberberine in rabbit]. AB - AIM: To explore the biotransformation of compound 7-(4-chlorbenzyl)-7,8,13, 13a tetrahydroberberine in the rabbit. METHODS: Analyze the rabbit bile sample with HPLC, LC/MS and LC/NMR. RESULTS: A metabolite and unchanged 7-(4-chlorbenzyl) 7,8,13,13a-tetrahydroberberine were found in the rabit bile, the metabolite was characterized and its structure was elucidated. CONCLUSION: Compound 7-(4 chlorbenzyl)-7,8,13,13a-tetrahydroberberine is metabolized by demethylation at 10 OCH3 position. PMID- 12579884 TI - [Transdermal microparticle delivery by a supersonic-Helios gun system]. AB - AIM: To investigate the effect of particle size and high speed flow of helium gas on the systemic absorption of indomethacin using a needle-less injection system. METHODS: Poly-L-lactic acid microspheres containing indomethacin was prepared by the o/w solvent evaporation technique. After anesthetizing the male hairless rat, microspheres filled in the tube cartridge was accelerated by a stream of helium gas at various velocity in the Helios gun system, and then was introduced to the abdominal skin. RESULTS: Introduction of indomethacin to the hairless rat skin was proportionally increased with enhancing the helium pressure (supersonic flow). Bioavailability and Cmax were also dependent on the helium pressure. CONCLUSION: This method can be used to deliver the powered drug and/or microparticulate systems into the skin tissues and the systemic circulation. PMID- 12579885 TI - [Predicting skin permeability of drugs with theoretical parameters]. AB - AIM: To predict skin permeability of drugs with theoretical parameters. METHODS: The semiempirical self-consistent field molecular calculation AM1 method is utilized to obtain the structural parameters of drug molecules. Stepwise multiple regression analysis or BP network is then utilized to establish the correlation between skin permeability of drugs and their structural parameters. RESULTS: The calculated human skin permeability coefficients (kp) of 22 model drugs in vitro or the R values (R = absorbed/unabsorbed) of 17 drugs in vivo are in good agreement with their observed values. CONCLUSION: Theoretical parameters can be used to predict skin permeability of drugs. PMID- 12579886 TI - [Protective effect of ONO-1078, a leukotriene antagonist, on focal cerebral ischemia in mice]. AB - AIM: To determine whether ONO-1078 [pranlukast, 4-oxo-8-[p-(4-phenylbutyloxy) benzoyl-amino]-2-(tetrazol-5-yl)-4H-1-benzopyran hemihydrate], a potent leukotriene antagonist, has protective effect on focal cerebral ischemia in mice. METHODS: Focal cerebral ischemia was induced by permanent middle cerebral artery (MCA) occlusion in mice. ONO-1078 (0.01, 0.05, 0.10 mg.kg-1), dexamethasone (0.5 mg.kg-1), nimodipine (0.2 mg.kg-1) or saline (control) were injected i.p. once daily for 3 days, and 30 min before MCA occlusion. Twenty-four hours after cerebral ischemia, the neurological scores were evaluated, infarct volumes and areas of the right and left cerebral hemispheres were measured by computer imaging analysis. RESULTS: ONO-1078, dexamethasone and nimodipine reduced the neurological scores. ONO-1078 and dexamethasone reduced the ratio of right/left hemisphere area, indicating inhibition of brain edema, while nimodipine showed no effect. ONO-1078 dose-dependently reduced infarct size, and dexamethasone and nimodipine showed the same effect. CONCLUSION: ONO-1078 showed protective effect on focal cerebral ischemia. This may represent a novel approach to the treatment of acute cerebral ischemia. PMID- 12579887 TI - [Conjugation of mitoxantrone-loaded nanospheres and anti-C-erbB-2 monoclonal antibodies]. AB - AIM: To improve the treatment efficacy of anti-tumor drug mitoxantrone, the conjugation of mitoxantrone-loaded nanospheres and anti-C-erbB-2 monoclonal antibodies were prepared. METHODS: Mitoxantrone-loaded nanospheres were prepared with emulsion-heating solidification technique. A heterobifunctional reagent, N succinimidyl 3-(2-pyridyldithio) propionate (SPDP), was used as the crosslinker of mitoxantrone-loaded nanospheres and anti-C-erbB-2 monoclonal antibodies; pharmaceutical properties of immunonanocapsuls were studied; the conjugates of nanospheres and monoclonal antibodies was confirmed with immunological methods such as slide agglutination test, fluorescent immunossay and rosset formation test, fluorescent staining and scanning electron microscope. RESULTS: Mitoxantrone-loaded nanospheres were spherical, with smooth surface and median diameter of 0.665 micron. When stored at 3-5, 20-25 and 37 degrees C, RH 75% for three months, the appearance, morphology, size distribution, drug loading and in vitro release characteristics showed no significant change and the stability was satisfactory. The size analysis demonstrated that there was no obvious increase in the particle size of nanoparticles after conjugation. Immunological tests indicate highly selective binding of antibody-targeted nanospheres to C-erbB-2 overexpressing cells SK-BR-3. CONCLUSION: The conjugation of mitoxantrone-loaded nanospheres and anti-C-erbB-2 monoclonal antibodies can keep the activity of anti C-erbB-2 and increase the therapeutic efficacy of anti-mammary cancer drugs. PMID- 12579888 TI - [Natural anti-tumor compounds with activity of promoting microtubule assembly]. PMID- 12579889 TI - [Effect of puerarin on learning-memory behavior and synaptic structure of hippocampus in the aging mice induced by D-galactose]. AB - AIM: To study the effects of puerarin on learning-memory behavior of aging mice induced by D-galactose and its possible mechanism of action. METHODS: The aging mice were induced by s.c. 0.12 g.kg-1 D-galactose for 6 weeks. The aging mice were treated with three doses of puerarin once a day for 5 weeks. The spontaneous behavior and the learning memory behavior were tested in the aging mice using open field and Y-maze at the next day after the last treatment. The structure of Gray I synaptic interface in the CA3 area of the hippocampus was quantitatively analyzed by electronic microscope and computer image processing appliance. RESULTS: Compared with the D-galactose control group, puerarin (60 mg.kg-1) was shown to increase significantly the spontaneous behavior and explorative response in the open field, and improve remarkablely the learning and memory ability of the aging mice induced by D-galactose. Meanwhile, the thickness of post-synaptic density (PSD) was increased, and the width of the synaptic cleft in the hippocampus CA3 area was shortened. CONCLUSION: Puerarin showed an improvement effect against the memory impairment in the modelling aging-mice induced by D galactose. A pathological alteration of synaptic interface structure in hippocampus of the mice may be involved in the effect. PMID- 12579890 TI - [Effects of praeruptorin C on cell hypertrophy, intracellular [Ca2+]i, nitric oxide and signal transduction in isolated hypertrophied rat smooth muscle cells induced by angiotensin II]. AB - AIM: To investigate the effects of praeruptorin C (Pra-C) on smooth muscle cell (SMC) hypertrophy, intracellular calcium ([Ca2+]i), nitric oxide (NO) content and influence on cellular signal transduction in isolated cultured rat smooth muscle cell (SMC). METHODS: Hypertrophied smooth muscle cells (HSMCs) were induced by angiotensin II (Ang II), cell area was measured under inverted microscope. Nitric oxide (NO) concentration was measured using Griess method. [Ca2+]i was measured using Fura-2/AM. The responses to [Ca2+]i elevation stimulated by KCl (60 mmol.L 1 or norepinephrine (10 mumol.L-1) were observed by incubation with phorbol 12 myristate 13-acetate (PMA), staurosporine (ST), the agonist and inhibitor of protein kinase C (PKC), and pertussis toxin (PTX), the sensitive toxin of Gi. RESULTS: The cell area of SMCs were decreased by 39.01% (P < 0.001) and NO content of SMCs were significantly increased in Pra-C + Ang II group. In presence of 60 mmol.L-1 KCl or 10 mumol.L-1 NE, [Ca2+]i of SMCs in Pra-C + Ang II group was significantly decreased than that of Ang II group (P < 0.001) and closed to the normal group. Incubation of SMCs with PMA, ST and PTX, [Ca2+]i of SMCs in Ang II group was increased by PMA and decreased by ST and PTX, but that of Pra-C + Ang II group was similar to the normal group. CONCLUSION: These findings suggest that Pra-C can reduce vascular hypertrophy in isolated rat HSMCs, and this is associated with improvement of SMCs [Ca2+]i level, NO content and cellular signal transdution of PKC and Gi. PMID- 12579891 TI - [Study on vasorelaxations of oxyphenamone and its mechanism]. AB - AIM: To study the vasorelaxation action of oxyphenamone (Oxy) and its mechanism. METHODS: The contractile response of isolated rabbit renal, femoral and mesentery artery preparations was determined. RESULTS: Oxy was shown to inhibit the contractile force of renal, femoral and mesentery arteries induced by phenylephrine in a concentration dependent manner. The vasorelaxation produced by Oxy was not attenuated by removal of the endothelium. Oxy (10(-6)-10(-4) mol.L-1) relaxed the contractions induced by KCl 30 mmol.L-1 as well as KCl 80 mmol.L-1, but the contraction curve of KCl 80 mmol.L-1 was shifted significantly to the right. Oxy in lower concentration (10(-6) and 5 x 10(-6) mol.L-1) increased the contractions induced by Ang II, and in middle concentration (10(-5) mol.L-1) it did not affect the contractions induced by Ang II. Whereas in higher concentration (5 x 10(-5) mol.L-1) it obviously inhibited the contractions induced by Ang II. CONCLUSION: Oxy showed significant vasorelaxation to various vascular preparations, and its vasorelaxation action is endothelium independent. The mechanism of its vasorelaxations seems to be related with Ca2+ activated K+ channel (Kca channel) and Ca2+ channel in vascular smooth muscle cells but its true mechanism needs further study. PMID- 12579892 TI - [Effects of clenbuterol on nitrogen metabolism and G6PDH activity of rat hepatocyte]. AB - AIM: To study the effects of beta 2-adrenergic receptor-selective agonist clenbuterol on nitrogen metabolism and glucose-6-phosphate dehydrogenase activity of rat hepatocyte and its pharmacological mechanism. METHODS: Biochemical methods were used to study the influence of clenbuterol on urea-nitrogen concentration of hepatocyte culture medium, 3H-leucine incorporation into hepatocyte, insulin-like growth factor I (IGF-I) production and glucose-6-phosphate dehydrogenase (G6PDH) activity of rat hepatocyte. RESULTS: The results showed that urea-nitrogen production by cultured rat hepatocytes was markedly affected with clenbuterol treatment (1 x 10(-6) mol.L-1), urea-nitrogen concentration of culture medium was decreased by 25.51% (P < 0.05) compared with control. The inhibitory effect of hepatocyte urea-nitrogen production of clenbuterol was blocked by propranolol, a beta-adrenoreceptor antagonist (1 x 10(-6) mol.L-1), but hepatocyte urea-nitrogen level was not affected with propranolol treatment only (P > 0.05). The content of 3H-leucine incorporation in rat hepatocyte was significantly increased by 23.35% (P < 0.05) with clenbuterol-treatment (1 x 10(-6) mol.L-1), and the enhanced effect of 3H-leucine incorporation into hepatocyte was antagonized by propranolol (1 x 10(-6) mol.L-1. The level of 3H-leucine incorporation of rat hepatocyte was not influenced by propranolol alone. IGF-I production of rat hepatocyte might be affected by clenbuterol. IGF-I concentration of culture medium was increased by 39.46% with clenbuterol (1 x 10(-6) mol.L-1), but no significant difference was found compared with the control (P > 0.05). Moreover, G6PDH activity of rat hepatocyte was significantly decreased by 43.36% (P < 0.05) with clenbuterol treatment (1 x 10(-6) mol.L-1), and the declined effect of clenbuterol was antagonized by propranolol. G6PDH activity of rat hepatocyte was not affected on condition that propranolol was administered alone (P > 0.05). CONCLUSION: It is suggested that clenbuterol may regulate nitrogen and fat metabolism by means of increasing nitrogen retention and protein synthesis, and decreasing G6PDH activity of rat hepatocyte for pharmacological effects. PMID- 12579893 TI - [Effects of clonidine on myocardial Gs alpha mRNA expression after scalds in rats]. AB - AIM: To explore the role of clonidine (Clo) on myocardial Gs alpha mRNA expression after scalds in rats. METHODS: A 30% skin-full-thickness scald was produced by immersing rats in 95 degrees C water for 10 s. The myocardial Gs alpha mRNA expression level, cyclic AMP content and adenylyl cyclase (AC) activity were determined with dot blotting hybridization, in situ hybridization, radioimmunoassay and indirect method. RESULTS: Three hours after scalds, the myocardial Gs alpha mRNA was significantly decreased to (61 +/- 20)% of the control group (P < 0.01). AC activity and cAMP content were also decreased. Clo (0.3, 1.0 and 3.0 mg.kg-1, i.p.) was shown to increase myocardial Gs alpha mRNA expression level (P < 0.01 or P < 0.05) after scalds to (131 +/- 28)%, (142 +/- 51)% and (139 +/- 48)% of the scald group, respectively, which were correlated with the Clo dose (gamma = 0.597, P < 0.05). Clo 1.0 mg.kg-1 and 3.0 mg.kg-1 (i.p.) promoted AC activity and increased cAMP content, but Clo 0.3 and 0.1 mg.kg 1 showed no significant effect (P > 0.05). Selective I1-imidazoline receptor antagonist efaroxan (Efa) (10, 5 mg.kg-1, i.p.) was found to partially reverse the effect of Clo, while Efa 2.5 mg.kg-1 showed no significantly influence. The reduced quantity of Gs alpha mRNA expression level correlated well with the Efa dose (gamma = 0.900, P < 0.05). The change of AC and cAMP was similar to Gs alpha mRNA. CONCLUSION: Clo increased the myocardial Gs alpha mRNA expression, AC activity and cAMP content after scalds in rats. PMID- 12579894 TI - [Induction of CuZn-SOD mRNA expression and activity by PGMS in rat liver]. AB - AIM: To study the effect of propylene glycol mannate sulfate (PGMS) on induction of CuZn-SOD. METHODS: Wistar rats were given PGMS p.o. at different doses (0, 18.9, 37.8 and 75.6 mg.kg-1.d) for ten days. Then the rats were sacrificed and the total RNA was extracted from the livers. The total RNA samples were loaded on a 1% agarose gel to detect the quality of total RNA. RT-PCR was applied to study the expression of CuZn-SOD mRNA in rat livers. The amplified products were detected by the 1.5% agarose gel electrophoresis. Simultaneously, the CuZn-SOD activities in rat liver were determined by nitrite method. RESULTS: The total RNA extracted from rat livers was integrated without being decomposed by RNase. The level of CuZn-SOD mRNA of the high-dosage group (75.6 mg.kg-1.d) was higher than that of the control group (0 mg.kg-1.d) (P < 0.01); the CuZn-SOD activities of the high-dosage group were significantly higher than those of the control group (P < 0.001) and the CuZn-SOD activities of the middle- (37.8 mg.kg-1.d) and low dosage groups (18.9 mg.kg-1.d) were higher than those of the control group (P < 0.01). CONCLUSION: PGMS can increase the CuZn-SOD activities as well as CuZn-SOD on mRNA level. Therefore, it is possible for PGMS to counteract Atherosclerosis (AS) by inducing the expression of CuZn-SOD. PMID- 12579895 TI - [Synthesis and anti-inflammatory analgesic activities of phenylfuroxan-coupled diclofenac]. AB - AIM: To search for new derivatives of diclofenac (DC) having higher potency than the parent drug and lacking its undesirable effects. METHODS: Coupling DC with NO donor 3-hydroxymethyl-4-phenylfuroxan and its isomer through esterification, evaluating anti-inflammatory and analgesic activities, observing side effects in the rat gastrointestinal (GI) tract and assessing NO releasing ability both in vitro and in vivo. RESULTS: Fifteen new compounds including nine target ones (II1 9) were synthesized, and their structures were determined by IR, 1HNMR, MS and elemental analysis. Compounds II3 and II9 showed anti-inflammatory activity comparable to DC. Compound II2 showed stronger anti-inflammatory and analgesic activities and less GI side effect than DC, and released NO in vivo. CONCLUSION: Compound II2 is worthy to be intensively studied. PMID- 12579896 TI - [Anti-gastric ulcer sesquiterpene lactone glycosides from Crepis napifera]. AB - AIM: The anti-gastric ulcer constituents from the roots of Crepis napifera (Franch) Babc (Compositae) were studied. METHODS: Solvent partition, Si gel and Rp-18 column chromatography, crystallization and spectral methods were used to extract, isolate and identify two compounds. The activity of compound 1 was tested on the rat stomach by determining the effect on aspirin-induced gastric lesions and on histamine-stimulated gastric acid secretion. RESULTS: Two sesquiterpene lactone glycosides, taraxinic acid-1'-O-beta-D-glucopyranoside (1) and 11,13-dihydro-taraxinic acid-1'-O-beta-D-glucopyranoside (2) were obtained. Compound 1 at the dose of 80 mg.kg-1 p.o. inhibited significantly the development of aspirin-induced gastric lesions in the rat and at an i.v. dose of 70 mg.kg-1 did not affect histamine-stimulated gastric acid secretion in the lumen-perfused rat stomach. CONCLUSION: Compound 1 is the active component of the plant which protects gastric mucosa and exhibits anti-gastric ulcer action. PMID- 12579897 TI - [Chemical constituents of Notoseris rhombiformis]. AB - AIM: To investigate the chemical constituents from the whole plant of the endemic plant Notoseris rhombiformis Shih. METHODS: Column chromatography was used for separation and purification, while spectral analysis was used for structure elucidation. RESULTS: Eight compounds were isolated from the whole plant of this Chinese endemic plant and their structures were elucidated as notoserolide A (1), notoserolide D (2), austricin (3), jacquilenin (4), 3 beta, 14-dihydroxy-11 beta, 13-dihydrocostunolide (5), p-hydroxyphenylacetic acid (6), luteolin-7-O-beta-D glucopyranoside (7) and daucosterol (8), respectively. CONCLUSION: All compounds were isolated from this plant for the first time. Among them, 2 is a new compound. PMID- 12579898 TI - [P53 gene mutation detection by bioluminometry assay]. AB - AIM: To develop a simple, fast and inexpensive approach as well as an instrument for detection of gene mutation. METHODS: Pyrosequencing based on bioluminometry assay was employed to detect gene mutation. Pyrosequencing is a method of sequencing by synthesis step-by-step using four enzymes, DNA-polymerase, ATP sulfurylase, luciferase and apyrase. The signal was produced by detecting pyrophosphate released during a dNTP incorporation. For mutation detection, a DNA fragment was amplified by PCR at first, followed by a single-stranded DNA preparation. In the second step, a short primer was annealed to the position just before the mutation point. Finally, specific dNTPs were added in terms of the template sequence. The mutation species can be readily determined by the sequence. RESULTS: A new instrument was developed for gene mutation detection by pyrosequencing. To iteratively inject small amount of each dNTP for the sequencing reaction, capillaries were used to connect dNTP reservoirs and the reaction chamber. Each dNTP was delivered by adding a gas pressure on the top of a corresponding dNTP reservoir, by which 0.2 microL of dNTP can be exactly added each time. It was theoretically proved that undesired liquid seep through the capillary did not affect the sequencing reactions in pyrosequencing. In addition, the three possible variants (wildtype, mutant and heterozygote) of a mutant point Cys275Ser in P53 gene exon 8 were determined by pyrosequencing using the instrument. A simple method was also described for rapidly distinguishing the type of a variant. CONCLUSION: The developed method is very simple, and the corresponding instrument is inexpensive and easy to operate, which can be used to detect many types of mutation. PMID- 12579899 TI - [Detection of crystal polymorphs of nateglinide by DSC]. AB - AIM: To establish the differential scanning calorimetric (DSC) methodology for controlling the crystal-type B form of nateglinide. METHODS: Accurately weighed pure dried (P2O5 as desiccant for 4 h at 80 degrees C in vacuum) fine powder of crystal-type B and H of nateglinide were measured dQ/dT by DSC at heating rate of 10 degrees C. min-1 and temperature between 100 degrees C and 200 degrees C to calculate the enthalpy delta HB and delta HH. Accurately weight a series of uniform mixtures of crystal-type B and H of dried fine powder of nateglinide in different proportions. The enthalpy of the mixtures is measured by DSC as above to calculate the enthalpy (sigma delta H). Using B% as X, sigma delta H as Y, the regression equation was obtained. According to this equation, the unknown composition of mixed crystal was evaluated by the sigma delta H values. The method was used to control the limitation of crystal-type B of nateglinide by the sigma delta H value of mixture of known composition as reference. RESULTS: The results measured from different laboratories showed that the repeatability was 0.61% and the recoveries were 86.2%-127% when the amounts of crystal-type B were between 0-15%. CONCLUSION: This method can be used to evaluate the crystal-type B composition of nateglinide. PMID- 12579900 TI - [Study on the chiral separation of securinine by high-performance capillary electrophoresis and its stereoselective metabolism in rat]. AB - AIM: To establish a high-performance capillary electrophoresis (HPCE) chiral separation method for d-securinine and l-securinine, and use this method to investigate the stereoselective metabolism process of d- and l-securinine in Wistar rats. METHODS: The electrophoretic condition and parameters were investigated and the optimized conditions were as following: the electrophoretic medium was 40 mmol.L-1 Tris-H3PO4 buffer (pH adjusted to 6.0 with H3PO4) containing 32 mmol.L-1 HP-beta-CD as chiral selector. Determination was carried out with a UV detector at 254 nm. The separations were performed at 16 degrees C with a positive voltage of 15 kV. Samples were injected into the capillary by pressure for 6 s. The biological samples (urine, bile, plasma and feces) of rats were alkalized and extracted with ethyl acetate. RESULTS: The experimental results showed that the concentration of HP-beta-CD, the concentration of the running buffer and the pH value of the buffer were the main important factors which effected the resolution. d-Securinine and l-securinine were separated at baseline level under the determination conditions. The determination was not interfered by endogenous components and metabolites. After i.p. administration, the rats excreted more d-securinine than l-securinine through bile, urine and feces. The metabolism process in rats was stereoselective. CONCLUSION: This method is simple, reliable and suitable for studying the stereoselective metabolism of securinine in rats. PMID- 12579901 TI - [In vitro and in vivo study of two kinds of long-circulating solid lipid nanoparticles containing paclitaxel]. AB - AIM: To prepare long-circulating solid lipid nanoparticles containing paclitaxel with stearic acid, and investigate the in vitro and in vivo characterization of nanoparticles. METHODS: The method of "emulsion evaporation-solidification at low temperature" was used to prepare the stearic acid solid lipid nanoparticles containing paclitaxel. Its morphology was examined by transmission electron microscope. The HPLC method for determination of paclitaxel in nanoparticles or serum samples was established. The release of paclitaxel in vitro and the pharmacokinetics after i.v. bolus injection to mice were studied. RESULTS: The mean diameter of Brij78-SLN and F68-SLN is (103.5 +/- 29.2) nm and (220 +/- 98) nm, respectively. The nanoparticles release paclitaxel slowly and linearly, within 24 h, Brij78-SLN and F68-SLN release 8% and 20% of total drug, respectively. Long-circulation nanoparticles was found to stay in the blood circulation, with T 1/2 beta 10.1 h of F68-SLN, and T 1/2 beta 4.88 h of Brij78 SLN more than one commercialized paclitaxel injection, T 1/2 beta 1.3 h. CONCLUSION: Stearic acid might be a new drug carrier material in the future. PMID- 12579902 TI - [Studies on the preparation of camptothecin niosomes]. AB - AIM: To study the non-ion surfactant vehicle (niosome) entrapped-camptothecin. METHODS: The niosome loaded with camptothecin was prepared from Span and cholesterol using aqueous dispersion of film. The vehicles were visualised by transmission electron microscopy and sized by laser particle analyzer on a Malvern Mastersizer. An HPLC analysis method of the camptothecin was established by fluorescence detection. The entrapment efficiency of the niosomes containing camptothecin was determinated after the ultracentrifugation of the niosome. The antitumor activities of the vehicles on S180 sarcoma in mouse were studied. RESULTS: The given niosomes were the suspension finely dispersed in aqueous solution. They were spherical vehicles with the single lamellar bilayers similar to phospholipid vehicles. The mean sizes of the vehicles were (565 +/- 6) nm. The recovery of the HPLC analysis method was 100.3% with 0.4% RSD. The entrapment efficiency of the camptothecin encapsulated by the niosome was 61%. The inhibition (%) of the niosome loaded with camptothecin on S180 sarcoma in mouse were 76.1% (P < 0.05). After the given dose the weight of the mouse of the niosome groups were 92.7% (P > 0.05) and 134.7% of blank control groups and compatothecin solution groups, respectively. CONCLUSION: The camptothecin niosomes were spherical in shape and similar to phospholipid vehicles with singlelamellar bilayers. Their size distributions were narrow. Their entrapment efficiency were higher. Its antitumor activity was better than camptothecin. PMID- 12579903 TI - [matK and its nucleotide sequencing of crude drug chuanxiong and phylogenetic relationship between their species from China and Japan]. AB - AIM: To provide more molecular evidences for species relationship between Chuanxiong (Ligusticum chuanxiong Hort.) from China and Japanese Chuanxiong (Senkyu in Japanese) (Cnidium officinale Makino). METHODS: To sequence such two genes as internal transcribed spacer (ITS) from nuclear rDNA and maturase for lysine (matK) in tRNA(lys) (UUU) intron from chloroplast DNA of both Ligusticum chuanxiong and Cnidium officinale using PCR direct sequencing and to analyze the sequence variation of two genes between these two species. RESULTS: The matK gene sequence of Ligusticum chuanxiong and Cnidium officinale is 1268 bp in length, coding 422 amino acids of maturase protein. ITS gene sequence 699 bp, consisting of 54 bp of 18S rRNA-3', 215 bp of ITS1, 162 bp of 5.8S rRNA, 222 bp of ITS2, 46 bp of 26S rRNA-5'. Multiple sequence alignment shows that the sequence of two genes between dried crude drug and fresh voucher material of Ligusticum chuanxiong and Cnidium officinale, there is 1 variable site (T-->C) in matK (upstream at 595 nt) and ITS (ITS1 at 54 nt) between Ligusticum chuanxiong and Cnidium officinale. CONCLUSION: Based on homology analysis of two genes plastid matK and nuclear ITS, the origin of Chuanxiong from China and Japan ought to be identical, the scientific name Cnidium officinale of Japanese Chuanxiong should be changed to Ligusticum chuanxiong. PMID- 12579904 TI - [Spectral characteristics, biogenesis and biological activities of oligostilbenes]. PMID- 12579905 TI - [Research development on the enantiomeric separation of drugs using non cyclodextrin capillary electrophoresis]. PMID- 12579906 TI - [Shunt tube implantation combining amniotic membrane transplantation and implantation of Molteno implant for glaucoma after penetrating keratoplasty]. AB - PURPOSE: To investigate the therapeutic effect of aqueous humor shunt implants with amniotic membrane transplantation on intractable glaucoma. METHODS: Glaucoma was induced in rabbits by the injection of alpha-chymotrypsin into the posterior chamber of the eyes. The rabbits were divided into four groups. Group A: control group, Group B: single shunt tube group, Group C: shunt tube with amniotic membrane transplantation group, Group D: shunt tube with amniotic supporter and amniotic membrane transplantation group. The intraocular pressure(IOP), histology and filterable ability of the tissue around the tubes were studied. The therapeutic effect of the three methods for the glaucoma was compared. From 1998 to 1999, 42 eyes of 41 patients with uncontrolled glacoma after penetrating keratoplasty were randomly assigned into two groups. One group (12 eyes) underwent implantation of shunt tube combining transplantation of amniotic membrane. The other group underwent implantation of a single plate Molteno implant. Clinical records were reviewed to ascertain postoperative IOP, visual acuities, number of medications. RESULTS: The IOP elevated after the operation and reached at the peak on the third day for all groups and then dropped slowly. The IOP was 33.34 +/- 5.54 mmHg (1 mmHg = 0.133 kPa) for Group A and 27.88 +/- 8.86 mmHg for Group B three months after the operation. There was no statistical difference between the two groups (P = 0.274). The IOP was 22.33 +/- 3.73 mmHg for Group C and there was statistical significant difference between Group C and Group A (P = 0.02) and no difference between Group C and Group B (P = 0.113). The IOP was 15.74 +/- 2.94 mmHg for Group D and there was statistical significant difference between Group D and Group A (P = 0.001) and Group B (P = 0.036). There was no difference between Group D and Group C (P = 0.09). The study of horseradish peroxidase penetrability indicated that there was peroxidase in the tissues around the tube with amniotic membrane transplantation and no peroxidase for simple shunt tube. The fibrous tissue near the tube was denser in simple shunt tube group than that in containing amnion groups. In clinic, the basic data existed no statistical difference between the two groups before surgery. The IOP was 54.42 +/- 9.65 mmHg in shunt tube with amnion group and 43.28 +/- 10.57 mmHg in simple plate Molteno implant group (P = 0.535) before operation. There was significant relativity of the visual acuity before and after the operation in two groups (r = 0.916, P = 0.002 and r = 0.962, P = 0.000). Most patients of the two groups had to use one or two anti glaucoma drugs. The overall success rates were 58.3% for shunt tube with amnion group and 66.7% for Molteno implant group (P = 0.73) within the follow-up period. Kaplan-Meier success analysis indicated one year cumulative success rates of 52.25% for shunt tube with amnion group and of 65.3% for Molteno implant group (P = 0.42). CONCLUSION: Amniotic membrane can inhibit proliferation of the scar around the shunt tube. Shunt tube implantation conbining amniotic membrane transplantation can significantly lowered the IOP of glaucoma. Combined using amnion supporter can expend shunt area and significantly reduce IOP of glaucoma. The effect of shunt tube implantation combining amniotic membrane transplantation has the similar result of single plate Molteno implant for the glaucoma after penetrating keratoplasty. PMID- 12579907 TI - [Effect of amniotic membrane on filtering bleb after trabeculectomy in rabbit eyes]. AB - OBJECTIVE: To investigate the effects of amniotic membrane on filtering blebs after trabeculectomy. METHODS: Twelve rabbits underwent bilateral trabeculectomies. One eye was randomly sutured preserved human amniotic membrane between the sclera and the scleral flap and the other eye served as the control. The filtering blebs and filtering function were observed at the 1st, 2nd and 3rd week after the operation. The scar development situation between bleb and episclera was observed under the light microscope. RESULTS: The blebs in amniotic membrane group were eminence, effuse and had a good filtering function at the 1st, 2nd and 3rd week after the operation. The blebs in control group were cicatrized and had a bad filtering function. Histological observation showed no scar development, less fibroblast cell and more inflammatory cells infiltration at the site of trabeculectomy in amniotic membrane group. On the contrary, the control group showed more fibroplasia and fibroblast hyperplasia. CONCLUSION: Amniotic membrane can reduce the scar, improve filtering function. PMID- 12579908 TI - [Clinical observation on the effect of 4% pilocarpine gel used one dose per day in the treatment of glaucoma]. AB - PURPOSE: To compare the efficacy and safety of 4% pilocarpine gel(PG) and 1% pilocarpine solution(PS). METHODS: Twenty cases(40 eyes) with ocular hypertension or primary open angle glaucoma(POAG) in early stage were treated by 4% PG(once per night) and PS(4 times per day). We compared the side-effects and the changes in the intraocular pressure (IOP), pupil size and diopter of 2 groups after medication. Self-control was chosen. RESULTS: The effect of 4% PG on IOP can last 20 hours. The IOP decreasing range of the PG group was from 9.3% to 13.8%, while the PS group was from 12.4% to 16.2%. The decreasing range at 10 AM for four weeks of the PG group was from 11.3% to 17.6% while the PS group was from 17.4% to 19.7%. There is no statistically significant difference between two groups. There are more obvious changes in the pupil size and diopter in the PS group and less complain of blurred vision in the PG group. CONCLUSION: The IOP decreasing effect of 4% PG is comparable with that of 1% PS, but the 4% PG is better than 1% PS as concerning the patient compliance and the side effects. So the 4% PG can be used as a substitute for the 1% PS in the clinic. PMID- 12579909 TI - [The contrast study of preoperative flash visual evoked potential test and the postoperative resumed visual acuity in senile cataract patients]. AB - PURPOSE: To study the using value of flash visual evoked potentials (FVEP) in predicting the resumed visual acuities in senile cataract patients. METHODS: Forty-three eyes with senile cataract had been conducted extracapsular cataract extraction combined with posterior chamber intraocular lens implantation. The contrast study had been carried out in this research between preoperative FVEP, light projection and color discrimination tests with the postoperative regained visual acuities. RESULTS: There were significant correlations between the regained visual acuities and the amplitudes of FVEP (X1), the latencies of P1 wave (X2) as well as color discriminations (P < 0.01). The coefficients were 0.7277, -0.5678 and 0.4302 in sequence. There was no correlation between the resumed visual acuities and the light projection test before operation. The formula of linear regression between the visual acuities (Y) and the indexes of FVEP was Y = 0.2255 +0.047X1 -0.0026X2. CONCLUSION: FVEP is an effective method to evaluate the visual function before the cataract operation. It may predict the resumed visual acuity. PMID- 12579910 TI - [The clinical application of different brightness and different saturation D-15 tests]. AB - PURPOSE: To compare the clinical application of different brightness and different saturation D-15 tests. METHODS: Eighteen normal subjects (30 eyes), 19 cases (38 eyes) of congenital color vision defects and 36 cases (59 eyes) of eye diseases were tested with Panel D-15 test, Hann's double D-15 test, middle and low saturation CAS-PI (Psychological Institute, Chinese Academy of Sciences) D-15 tests. RESULTS: A few of the normal eyes made minor errors in the low saturation D-15 tests. All of the protanopes and deuteranopes could be detected correctly. Protanomalias and deuteranomalias showed normal arrange or some type of abnormal arranges in all of the 5 sets of D-15 tests. In sick eyes, the abnormal rates were the highest in the low saturation D-15 tests and the lowest in Panel D-15 test. CONCLUSION: The five sets of different brightness and different saturation D-15 tests had similar efficiencies of detecting congenital color vision defects. The abnormal rates of CAS-PI(4/5)D-15 tests and the CAS-PI(2/5)D-15 tests were similar to those of Panel D-15 test and Hann's Double D-15 test. Both tests can be used in the clinical setting. PMID- 12579911 TI - [Quantitative measurement of macular thickness in normal subjects by optical coherence tomography]. AB - OBJECTIVE: To measure the thickness of retinal neurosensory layer, retinal pigment epithelium (RPE) and choriocapillaris in normal subjects by optical coherence tomography (OCT). METHODS: Sixty normal subjects (120 eyes) were examined by OCT with 4 mm scanning length horizontally and vertically through foveola. The mean thickness of retinal neurosensory layer, RPE and choriocapillaris thickness in foveola, 175 microns and 750 microns to foveola, were calculated and compared. RESULTS: The mean thickness of retinal neurosensory layer is 146.34 +/- 8.58 microns in foveola; 155.58 +/- 10.20 microns in 175 microns to foveola and 252.98 +/- 14.07 microns in 750 microns to foveola. The mean thickness of RPE and choriocapillaris is 106.26 +/- 10.58 microns in foveola; 104.98 +/- 7.24 microns in 175 microns to foveola and 106.84 +/- 9.95 microns in 750 microns to foveola. The results show that there are no significant in mean thickness of retinal neurosensory layer in 175 microns to foveola in statistics, but there are differences in 750 microns to foveola between nasal and temporal or superior, inferior and nasal, temporal. The differences in mean RPE and choriocapillaris thickness of all sites are not significant. Correlative analysis showed that there is no correlation between retinal neurosensory layer, RPE and choriocapillaris thickness and age. CONCLUSION: OCT can accurately measure the thickness of retina in normal eyes and provide a subjective and Quantitative method to diagnose macular diseases. PMID- 12579912 TI - [The recent progress on the role of alpha-crystallin as a molecular chaperone in cataractogenesis]. AB - The alpha-crystallin is a major structural protein within the lens and consists of two types of subunit, alpha A and alpha B. These subunits propose a model for the quaternary structure of alpha-crystallin. It has been known for many years that its main function is to serve as a structural and refractive elements in lens. Until 1992, Horwitz suggested that alpha-crystallin could act in a chaperone-like manner. Many studies have showed that alpha-crystallin can protect enzymes and other crystallins against both chemically- and thermally-induced inactivation or aggregation, which may play an important role in maintaining transparency of lens. A polypeptide sequence of alpha A-crystallin or extension of alpha B-crystallin in C-terminal-region and hydrophobic N-terminal-region are all essential for chaperone function. A decrease in chaperone activity from nuclear crystallin has been showed in age-dependent fashion. Post-translational modifications occurring to alpha-crystallin with increasing age and during cataract formation may decrease the chaperone activity of alpha-crystallin to different extent, resulting in an increase in other crystallins aggregation and enzymes inactivation. Therefore, these aggregates will increase scattering of light and lead to lens opacification. Development of protective agents for molecular chaperone activity of alpha-crystallin, in conjunction with knowledge of decreasing post-translational modifications, could potentially lead to a new field for the research on pathogenesis and clinical treatment of cataract. PMID- 12579913 TI - [Phacoemulsification in eyes with Fuchs' heterochromic iridocyclitis]. AB - PURPOSE: To evaluate the effect and safety of phacoemulsification with intraocular lens (IOL) implantation in eyes with Fuchs' heterochromic iridocyclitis. METHODS: We reviewed the 13 patients (14 eyes) with heterochromic iridocyclitis who had phacoemulsification and IOL implantation. Intraoperative and postoperative complication was observed. RESULTS: Intraoperative complication included miosis and hemorrhage in anterior chamber. Postoperative complication were glaucoma and hyphemia. Best corrected postoperative visual acuity was 1.0 or better in 65% of eyes. CONCLUSION: Eyes with heterochromic iridocyclitis tolerated well to the phacoemulsification with IOL implantation. PMID- 12579914 TI - [Observation of anterior chamber angle after phacoemulsification and extra capsule cataract extraction and intraocular lens implantation]. AB - OBJECTIVE: To observe and compare the difference of anterior chamber angle after Phacoemulsification and Intraocular Lens Implantation (Phaco & IOL) through a tunnel incision and Extra Capsule Cataract Extraction and Intraocular Lens Implantation (ECCE & IOL). METHODS: The changes of internal incision, the peripheral iris and the fixation of lens intracapsular of 36 eyes were individually observed after the operation. RESULTS: Desemet's membrane detachments occurred in three eyes (8.35%) in the group of Phaco & IOL and four eyes (11.1%) in the group of ECCE & IOL. The difference was not significant. (P > 0.05) Peripheral anterior synechiae occured in three eyes (8.35%) of Phaco & IOL group and in 17 eyes (47.27%) of ECCE & IOL group. The difference was significant (P < 0.05). Intracapsular lens account for 31 eyes (86.1%) of Phaco & IOL group and ten eyes (22.5%) in ECCE & IOL group. The difference was statistically significant. (P < 0.05) CONCLUSION: Phaco & IOL can decrease the incidence of peripheral iris anterior synechiae and increase the chance of intracapsular placement of IOL. PMID- 12579915 TI - [The clinical analysis of 260 combined surgery of glaucoma and cataract]. AB - PURPOSE: To evaluate the efficacy of the combined surgery of glaucoma and cataract. METHODS: We reviewed 260 consecutive cases (279 eyes) who underwent the combined procedure of glaucoma and cataract. The patients were divided into three groups: group 1: extracapsular cataract extraction combined with trabeculectomy; group 2: extracapsular cataract extraction and intraocular lens implantation combined with trabeculectomy; group 3: cataract phacoemulsification and intraocular lens implantation combined with trabeculectomy; We analyzed and compared the postoperative visual acuity (VA), intraocular pressure(IOP) and complications among three groups. RESULTS: 16.2%, 32.4% and 43.5% of patients in three groups achieved 0.3 or better VA. The VA improved rate were 46.6%, 70.4% and 78.3% respectively. The mean postoperative IOP of three groups were 12.72 +/- 6.66 mmHg(1 mmHg = 0.133 kPa), 13.01 +/- 5.26 mmHg, 12.63 +/- 4.66 mmHg(compared with preoperative IOP, P < 0.01). The complications of group 3 were much less than those of the other two groups. CONCLUSION: The combined surgery can control the IOP effectively; The triple procedure can improve the VA rapidly and the phacoemulsification technique can decrease the complications. PMID- 12579916 TI - [Surgical techniques to deal with lens nucleus' fragment of posterior capsular rupture during the phacoemulsification]. AB - OBJECTIVE: To investigate some surgical techniques in dealing with a impending dropped nucleus when posterior capsular rupture during the phacoemulsification. METHODS: The postoperative results of 18 cases (18 eyes) whose posterior capsular of lens ruptured with a impending dropped nucleus during the phacoemulsification were studied retrospectively. Three surgical techniques to prevent lens nucleus' fragments dropping into the vitreous cavity were applied. They are consecutive phacoemulsification with lower flowing rate and lower irrigation, injection of viscoelastic substance with high elasticity and anterior segment vitrectomy with no irrigation. The follow-up was 12-24 months (average, 18.2 months). RESULTS: Except for a special case, no lens nucleus' fragments dropped into the vitreous cavity. Posterior chamber intraocular lens(IOL) was implanted primarily in all cases (18 eyes). Best corrected visual acuity was above 0.5 in 15 eyes (83.3%). The main complications included corneal edema and cystoid macular edema. CONCLUSION: With safety, effectiveness and less complications, these three surgical techniques are suitable to deal with different types of impending dropped lens nucleus' fragments during the phacoemulsification. It is advisable to apply these techniques in clinical practice. PMID- 12579917 TI - [Vitrectomy combined with intraocular lens implantation for 103 selected cases of complicated ocular trauma]. AB - PURPOSE: To observe the visual outcome of vitrectomy combined with intraocular lens (IOL) implantation in one operation for patients with complicated ocular trauma. METHODS: Clinical records were retrospectively reviewed in 103 patients with complicated ocular trauma (104 eyes) who underwent vitrectomy combined with IOL implantation in one operation. RESULTS: The series of cases included 35 patients with intraocular foreign body (IOFB), 37 with penetrating injury, 9 (10 eyes) with explosive injury but without IOFB, and 22 with blunt trauma. Preoperatively, visual acuity was no-light-perception to counting fingers in 88 eyes (87.1%) of 100 patients (101 eyes). Visual acuity improved in 94 eyes (93.1%) including 74 eyes (73.3%) with 0.1-1.5 postoperatively. Retinal detachment occurred in one eye during follow-up and cured with pars plana vitrectomy. CONCLUSION: Vitrectomy combined with IOL implantation in one operation can be a safe, fast and effective approach for visual recovery in selected patients with complicated ocular trauma. PMID- 12579918 TI - [Clinical and histopathological studies of melanocytoma of the optic disc]. AB - OBJECTIVE: To examine the clinical features that may be useful in determining the malignant transformation of a melanocytoma of the optic disc. METHODS: The clinical documentation of 10 cases, including visual acuity, visual field, fundus findings and fluorescein angiogram were analyzed retrospectively. Three of them undergone histopathologic examinations. RESULTS: A benign melanocytoma may have some degree of decreased visual acuity, progressive nerve fiber bundle defect, certain amount of pigment dispersion into the vitreous, slight macular exudation and extension of the tumor tissue into the peripapillary area of the retina. CONCLUSION: The progression of the associated clinical symptoms and signs appears to be not necessarily indicative of a malignant transformation of the benign disease unless they are associated with rapid enlargement of the tumor mass as well as marked pigmentary changes of the tumor surface. PMID- 12579919 TI - [Radiological study of orbital foreign body]. AB - OBJECTIVE: To study the factors influencing of foreign body. METHODS: 527 cases of orbital foreign body was analysed and the relationship between foreign body and projection position or projection angle was retrospectively reviewed. RESULTS: Foreign body in 13 cases was found in the lateral radiogram(No foreign body in the posteroanterior radiogram). In 12 cases foreign body was found in the Belot's radiogram(No foreign body in P. A. and lateral radiognam). In ten cases foreign body was found in the Vogts radiogram (No foreign body in P. A., lateral, Belot's radiogram). In the first radiogram, one case was found to have foreign body in the Lateral but P. A. In the second radiogram, just the reverse of above. CONCLUSION: The image of foreign body was influenced by projection position and projection angle between foreign body and X-ray. The patients must take Belot's or Vogt's radiogram except routine P. A. and Lateral radiogram. Sometimes the patients should be taken more radiogram after moving the eye ball direction in order to find the small and flact foreign body. PMID- 12579920 TI - [Advancement in the study of embryonic stem cells and the emerging potential promising future of ocular tissue engineering]. AB - The advancement of the study in embryonic stem cells and tissue engineering makes it promising to effectively solve the intractable problems in the present ophthalmic practice. The problems include: no effective measures to regain the visual function of the patients with late stage glaucoma or retinal diseases, shortage of appropriate biocompatible donor tissues in treating ocular surface diseases. PMID- 12579921 TI - [Evaluation of polymerase chain reaction for detection of chlamydia trachomatis in eye swabs]. AB - OBJECTIVE: To investigate the clinical value of Polymerase Chain Reaction (PCR) in the diagnosis of trachoma. METHODS: Direct amplication of 16S rRNA gene which is chlamydia-specific by PCR was used to detect chlamydia trachomatis (CT). RESULTS: Of 93 subjects who conformed to the diagnosis criteria of trachoma, 69 (74.2%) was PCR-Positive; Of 178 subjects who did not comform to the criteia, 6(3.4%) was PCR-positive. PCR had a sensitivity of 74.2% and a speciticity of 96.7% for the clinical diagnosis of trachoma. The difference between trachoma group and nontrachoma group is significant(P < 0.001). CONCLUSION: The PCR may be a good method in the diagnosis of ocular chlamydia trachomatis infection because of its excellent sensitivity and specificity. PMID- 12579922 TI - [Morphological changes of cultured rabbit retinal cells after injury]. AB - PURPOSE: To investigate the morphological changes of cultured rabbit retinal cells after injury. METHODS: Mechanical scratch was done in nenatol rabbit retinal cell culture and in Schwann Cell(SC)/retinal cell colculture. The morphological changes of different sorts of cells after the injury were observed under the phase contrast microscope for 24 h. RESULTS: About 30 min after injury, many glial cells moved to the blank region and proliferated to form gliosis. Axons of retinal neurons began to regenerate 6 h post injury. The direction of regrowth was influenced by the environment. In the coculture, regeneration of axons was discerned only 3 h after injury. CONCLUSION: Axons of retinal neurons had strong capability of regeneration; Gliosis was not the absolute obstacle of regeneration; SC played a role in promoting the axon regeneration. PMID- 12579923 TI - [Culture and identification of human vascular endothelial cells]. AB - PURPOSE: To investigate the culture technigue of human vascular endothelial cells in vitro and improve the success rate of culture. METHODS: Human vascular endothelial cells were isolated from umbilical cords by "Irrigative digestion" technique with 0.2% collagenase III and were digested by 0.0625% tripsin when the cells confluent. The cells were identified by morphologic and immunohistochemical assays. RESULTS: The cultured human umbilical vein endothelial cells attached completely after 24 hours in vitro, reached confluence in 3-5 days with the typical appearance of "stone of pave road" and were passaged on time; Immunohistochemically these cells were stained positively by CD34 monoclonal antibody. CONCLUSION: "Irrigative digestion" technique was a reliable method to obtain human umbilical vascular endothelial cells with high success rate. It can be widely used to the culture and researches of human vascular endothelial cells. PMID- 12579924 TI - [The culture and cryopreservation of human conjunctival epithelium in vitro]. AB - PURPOSES: To detect the best method of culture and preservation of human conjunctival epithelia. METHODS: Human conjunctival epithelia were cultured by tissue inoculation, mechanical separation or enzyme digestion with 0.25% trypsin. The cultured cells were identified through their morphology, growing features and immunohistochemical staining. The third and fourth passage confluent cells were frozen in liquid nitrogen, and resuscitated 30 days later. RESULTS: Cells from tissue digested with 0.25% Trypsin grew much better than those from tissue inoculation. No cell outgrew in mechanical separating group. The cultured cells spread along the flask in polygonal shape and positive in pan-Keratin staining. Ninty percent of the cryopreserved cells were successfully resuscitated. CONCLUSION: Tissue digestion with trypsin is the best approach to culture human conjunctival epithelium, and the cultured cells can be cryopreserved in liquid nitrogen. PMID- 12579925 TI - [A histochemical and immunohistochemical study of cornea and adjacent bulbar conjunctiva in Mooren's ulcer]. AB - OBJECTIVE: To investigate immunopathologic changes of cornea and adjacent bulbar conjunctiva in Mooren's ulcer, and roles of macrophage, T lymphocyte, and VCAM-1 in the pathogeny of Mooren's ulcer. METHODS: The specimens of the cornea lesions and their adjacent bulbar conjunctiva taken from 18 active Mooren's ulcer cases who received resection of conjunctiva and lamellar keratoplasty surgery were studied with histochemical and labelled streptavidin biotin (LSAB) immunohistochemical techniques to investigate the expressions of CD68, CD3, CD4, CD8, CD1a, VCAM-1, HSP70, and TNF in the affected cornea and adjacent conjunctiva. RESULTS: Under the histochemical examination, infiltration of neutrophils, plasma cells, lymphocytes, and eosinophils were found in the affected cornea and conjunctiva. Degeneration and necrosis of corneal epithelium, Bowman's layer, collageneous fiber in stromal layer, and edema of conjunctiva were shown. Infiltration of a large number of macrophages and T lymphocytes in the affected cornea and adjacent conjunctiva were shown in LSAB immunohistochemical staining. The main infiltrating T lymphocytes were the CD4 positive cells. The CD4/CD8 ratio was significantly higher than that of normal contrals (P < 0.01). VCAM-1 was strongly expressed in the affected corneal and conjunctival epithelium. There were no significant differences for expressions of CD1a, HSP70, and TNF between the pathological specimens and normal contrals. CONCLUSION: The large number of infiltrating macrophages are responsible for the immunopathological damage caused by T lymphocyte infiltration, aberrant production of cytokikens and aberrant expression of HLA-DR antigen of Mooren's ulcer. There is an aberrant expression of VCAM-1 in the affected corneal and conjunctival epithelium. The role of VCAM-1 in the immunopathogenesis of Mooren's ulcer needs to be studied. PMID- 12579926 TI - [Analysis of refractive state after excimer laser photorefractive keratectomy in myopia]. AB - OBJECTIVE: To evaluate refractive state after excimer laser photorefractive keratectomy (PRK) in patients with myopia. METHODS: 234 cases(391 eyes) followed up for two years were divided into two groups, group I: mild and moderate myopia(< or = -6.00 D), 228 eyes; group II: High myopia(> -6.00 D), 163 eyes. The refractive state of patients at three, six, twelve and twenty-four months postoperatively were monitored. RESULTS: At two years after photorefractive keratectomy (PRK), refractive regression (> -0.5 D), overcorrection(> 1.00 D), undercorrection(> 1.00 D), postoperative residual astigmatism postoperative astigmatism and hyperopia shift in group I were 1.7%, 4.4%, 5.7%, 12.8%, 7.4% and 7.5% respectively, while in high myopia group were 15.9%, 4.9%, 23.3%, 29.7%, 31.1% and 6.7% respectively. CONCLUSION: The results prove that photorefractive keratectomy is an effective, precise, stable and safe method for correction of mild and moderate myopia, but insufficient for high myopia. PMID- 12579927 TI - [Clinical investigation of 0.3% levofloxacin eyedrops on the treatment of cases with acute bacterial conjunctivitis and bacterial keratitis]. AB - PURPOSE: To investigate the effects of 0.3% levofloxacin eyedrops on the treatment of acute bacterial conjunctivitis and bacterial keratitis. METHODS: The patients were randomly divided into treated group and control group. They were given 0.3% levofloxacin and 0.3% ofloxacin eyedrops respectively. RESULTS: From April to December in 1999, 132 cases were enrolled in the study. It showed that there was no significant difference between two drops in the cure and effective rates, but the mean cure day of keratitis in levofloxacin group was significantly shorter than that in ofloxacin group. No serious side effects occurred. CONCLUSION: Levofloxacin eyedrops was effective and safe antibiotics in treating bacterial conjunctivitis and keratitis. PMID- 12579928 TI - [Result of orthokeratology for treatment of young people with myopia]. AB - PURPOSE: To evaluate the effectiveness of orthokeratology (Ortho-K) for treatment of myopia in youngths. METHODS: 110 eyes of 56 young peoples with myopia received Ortho-K were studied. The patients were divided into 3 groups according to preoperative diopters. No. I: -1.00(-)-3.00 D, No. II: -3.25(-)-6.00 D, NO. III: 6.25(-)-7.50 D. The uncorrected visual acuities, residual diopters and corneal refractive powers at various time of three months after the operation were statistically analyzed and compared with that of preoperation. Correlation analysis and linear regression analysis were performed between the corneal refractive reduction (X) and clinical refractive reduction (Y) after 3 months of the operation. RESULTS: In 110 eyes, the uncorrected visual acuities in the first day, first week, first month, second month and third month after operation were significantly improved than that of the preoperation (P < 0.01). The mean residual diopters were significantly reduced than that of preoperation (P < 0.01). The mean refractive powers of cornea were significantly decreased than that of the preoperative (P < 0.01). There was significant correlation between the corneal refractive reduction and clinical refractive reduction. (r = 0.3181, P < 0.001). CONCLUSION: Orthokeratology is a safe and effective therapeutic method for treatment of myopia in youngths. The long term effect of Orthokeratology need further observation. PMID- 12579929 TI - [Chronopharmacology of tramadol in mice]. AB - AIM: To investigate the influence of dosing time on acute toxicity, action and pharmacokinetics of tramadol in mice to find the circadian rhythms of the drug. METHODS: Tramadol was given (i.p.) to mice at 08:00, 12:00, 16:00, 20:00, 24:00, 04:00 and the variations of their acute mortality, responses to hot plate and tail-pressing stimulation and pharmacokinetics were investigated. RESULTS: Significant circadian rhythm was found for the acute toxicity of tramadol in mice and the highest mortality was found when the drug was administered in the middle dark phase. The rhythm of drug toxicity coincided nicely with the rhythm of plasma drug level. Tramadol effect was demonstrated to vary circadianly and the highest effect occurred in the late dark phase. Dosing-time dependent kinetics of tramadol was also found in mice. CONCLUSION: Significant circadian rhythm of the acute toxicity, action and pharmacokinetics was found in mice when tramadol was given at different times of the day. PMID- 12579930 TI - [Effect and mechanism of securinine on synaptic transmission in the dentate gyrus of anesthetized rats]. AB - AIM: To study the effects of (+) and (-) securinine on the basic synaptic transmission and long-term potentiation (LTP) induced by high-frequency stimulation in the dentate gyrus of anesthetized rats and its possible mechanism. METHODS: The population spike (PS) was recorded by electrophysiological techniques. RESULTS: The results showed that at the concentration of 2 x 10(-9) mol.L-1, (+), (-) securinine showed no effect on either the basal PS or LTP. At high cocentrations (2 x 10(-8), 2 x 10(-7) mol.L-1), (+), (-) securinine not only potentiated the basal PS but also enhanced the magnitude of LTP. (+), (-) Securinine showed similar effects; and the effects were completely inhibited by the inhibitory amino acid GABA. CONCLUSION: These results suggest that the effects of (+), (-) securinine on induction and maintenance of LTP may be related to its blockade of GABA receptor. PMID- 12579931 TI - Synergistic effects of geldanamycin and antitumor drugs. AB - AIM: To study the effect of geldanamycin (GDM) on cell-cycle of human hepatoma BEL-7402 cells and the antitumor activity of cisplatin and mitomycin C in combination with GDM in vitro and in vivo. METHODS: MTT assay was used to determine the growth inhibition of hepatoma BEL-7402 cells. Cell cycle was analyzed by flow cytometry. Transplantable murine hepatoma 22 model was used to evaluate the antitumor activity of drugs in vivo. RESULTS: The IC50 value of GDM for hepatoma BEL-7402 cells by MTT assay was found to be 0.28 mumol.L-1. At concentrations of 0.1, 1.0, and 10 mumol.L-1, GDM reduced the proportion of S phase and induced G2/M arrest in BEL-7402 cells. At relatively low cytotoxic concentration, 0.1 or 0.2 mumol.L-1, GDM markedly potentiated the cytotoxicity of a series of chemotherapeutic agents including cisplatin, mitomycin C, adriamycin and cytarabine against BEL-7402 cells. The inhibition of tumor growth by cisplatin and mitomycin C was also enhanced in transplantable hepatoma 22-bearing mice when these agents were administered in combination with GDM 0.38 mg.kg-1. The synergistic effects were very significant with CDI < 0.7. CONCLUSION: These results suggest that GDM, as a biochemical modulator targeting Hsp90 function, may be potentially useful in cancer chemotherapy. PMID- 12579932 TI - [Studies on the anti-tumorpromotion activities of dehydroepiandrosterone and its mechanism of action]. AB - AIM: To investigate the anti-tumorpromoting activity of dehydroepiandrosterone (DHEA) and its mechanism of action. METHODS: Using croton oil-induced ear edema model and applying confocal laser scanning microscopy, flow cytometry, immuno fluorescent techniques to investigate the inhibitory effect of DHEA on tumor promotion. RESULTS: DHEA 25 mg.kg-1 was shown to inhibit croton oil induced ear edema in mice by 51%. DHEA at dose of 40 mg.kg-1 and 10 mg.kg-1 exhibited inhibitory effects on croton oil-induced ornithine decarboxylase (ODC) activity by 64% and 53%, respectively. These results revealed that DHEA can block the change of cell cycle and the percentage of S phase was decreased to 17% at concentration of 10(-7) mol.L-1. The increase of[Ca2+]i and pH as well as PKC activation induced by TPA stimulation were significantly inhibited by DHEA pretreatment. CONCLUSION: The present experiments demonstrate that DHEA appears to be an attractive candidate as an anti-tumorpromotion agent for tumor chemoprevention. The mechanism of its action might be related to its inhibitory effects on ODC activity and Ca(2+)-DG-PKC signal pathway. PMID- 12579933 TI - [Synthesis and antibacterial activity of 3-hydroxy-6-O-methylerythromycin-9-O substituted oxime derivatives]. AB - AIM: To study the antibacterial activity against erythromycin-resistant organisms of 3-hydroxy-6-O-methylerythromycin-9-O-substituted oxime derivatives, a new route of synthesis with 6 steps was designed. METHODS: The starting material, erythromycin A (1), was reacted with NH2OH.HCI to give 2, which reacted with BzBr to give 3. Selective methylation of C-6 hydroxy group using iodomethane afforded 4, which was hydrolyzed with loss of the 3-cladinosyl to give 5. Compound 5 was reduced by H2 to provide 6, which was treated with substituted benzyl chlorides to provide 7 and 8. RESULTS: Four unreported compounds (5-8) were synthesized. The antibacterial activity of the new compounds were tested in vitro against both erythromycin-susceptible and erythromycin-resistant organisms. The compounds 5 (MIC = 1 microgram.mL-1) and 6 (MIC = 1 microgram.mL-1) showed significant activity against Staphylococcus epidermidis 26,069 compared with erythromycin (MIC = 4 micrograms.mL-1). Compounds 5 (MIC = 16, 4 micrograms.mL-1), 7 (MIC = 32, 64 micrograms.mL-1) and 8 (MIC = 64, 32 micrograms.mL-1) showed better activity against Streptococcus pneumoniae 64 and Staphylococcus aureus 9525 than erythromycin (MIC > 128, 128 micrograms.mL-1). CONCLUSION: 3-hydroxy-6-O methylerythromycin-9-O-substituted oxime derivatives have stronger antibacterial activity against some erythromycin-resistant organisms than erythromycin A. PMID- 12579934 TI - ["Target secondary structural motif" in the action of antisense oligodeoxynucleotides]. AB - AIM: To optimize the antisense drug design based on the methods of secondary structure prediction of target mRNA by computer and the quantitative structure activity relationship analysis. METHODS: The secondary structures of mRNA were predicted by the software RNAstructure, then the antisense phosphorothioate oligodeoxynucleotides (AS-ODN) were designed against the secondary structural elements. The in vitro anti-tumor bioactivity of AS-ODN was evaluated by A549 lung carcinoma cell line. The multiple regression was performed with the computer program SPSS. RESULTS: AS-ODN with high bioactivity concentrated on some local secondary structural motifs that were composed of several secondary structural elements, designated here as "target secondary structural motif" (target motif). The target motifs were relatively stable in the whole mRNA structures, but there were one or more unstable secondary structural elements (free energy > 0) such as internal loops, knots, and hairpins, especially the bulge loops in the motif. Bioactivities of AS-ODN targeting different "target motifs" were statistically different (P < 0.01) while AS-ODN against the same "target motif" were with similar effects. CONCLUSION: The concept of the "target motif" was helpful for optimizing the antisense drug design and might be useful for discovering the local function of mRNA and designing oligonucleotide probes and primers. PMID- 12579935 TI - [Chemical constituents of Diuranthera inarticulata]. AB - AIM: To investigate the chemical constituents from the roots of Diuranthera inarticulata Wang et K. Y. Lang. METHODS: The compounds were isolated with silica gel column chromatography and their structures were identified on the basis of spectral data and TLC with authentic samples. RESULTS: From the methanolic extracts of the roots of Diuranthera inarticulata, ten compounds were obtained and nine of them were identified as chloromaloside A (III), diuranthoside A (IV), diuranthoside B (V), diuranthoside F (VI), diuranthoside G (VII), 2 alpha, 3 alpha, 19 alpha-trihydroxy-urs-12-en-28-oic acid (VIII), 3,6,7,3',4'-pentamethoxy flavone (IX), gitogenin (X) and daucosterol (XI). III-VI showed inhibition of growth with the yeast, Candida albicans. III, IV and VI also showed inhibition of growth with fungus, Aspergillus niger, in the test of antibacterial activity. CONCLUSION: All of these compounds were isolated from this plant for the first time. Among them, VI and VII are new steroidal saponins. PMID- 12579936 TI - [A new triterpene from the fruiting bodies of Ganoderma lucidum]. AB - AIM: To study the chemical constituents of the fruiting bodies of Ganoderma lucidum. METHODS: Individual constituents, isolated and repeatedly purified on silica gel column, were identified by physicochemical constants and structurally elucidated by spectral methods. RESULTS: From the alcohol extract, compound 2 was obtained and identified as 3 beta,7 beta-dihydroxy-4,4,14 alpha-trimethyl-11,15 dioxo-5 alpha-chol-8-en-24-oic acid. In addition, two known compounds, lucidenic acid A (1) and C (3) were obtained. CONCLUSION: Compound 2 is a new triterpene compound. PMID- 12579937 TI - [Physico-chemical properties and activity of glycoconjugate LbGp2 from Lycium barbarum L]. AB - AIM: To isolate and purify a glycoconjugate (LbGp2) from the fruit of Lycium barbarum L. and study its immunoactivity and antioxidative activity. METHODS: By means of gel permeation chromatography, LbGp2 was purified. Based on HPLC, CE, GC, SEC and component analysis and so on, its physico-chemical properties were studied. RESULTS: Molecular weights of LbGp2 was 68.2 ku and its carbohydrate content was up to 90.7%. Component analysis showed that it composed of Ara and Gal in a molar ratio of 3:4, and 18 kinds of amino acids. The immunologic function and bioactivity of Lbp2 has been studied preliminarily. Lbp2 was shown to increase rate of phagocyticaction and phagocytic index, promote lymphocyte translation and accelerate the production of serum hemolysin. LbGp2 has distinct effect of antioxidation and the superoxide anion produced by DMSO-NaOH system was scavenged effectively. CONCLUSION: LbGp2 was shown to be a kind of homogeneous glycoconjugate with good immunoactivity and antioxidative activity. PMID- 12579938 TI - [Metabolism of ginsenoside Rb1 and panaxadiol saponins by fungi]. AB - AIM: To study the metabolic process of ginsenoside Rb1 (G-Rb1) and panaxadiol saponins (PDS) by fungi. METHODS: Ten strains of fungi were incubated with G-Rb1 and PDS at a certain temperature with shaking. A portion was taken out at different time and mixed up with butanol. The butanol extract was analysed by thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and electrospray ionazition mass spectrometry (ESI-MS). RESULTS: It was found that there were ginsenoside-Rd (G-Rd), ginsenoside-F2 (G-F2), compound K (CK) and 20(S) protopanaxadiol (Ppd) metabolites beside the prodrug G-Rb1 induced by fungi (No. 1, 2, 3, 6, 8, 9). CONCLUSION: The six strains of fungi have different degrees of ability to metabolize G-Rb1 and PDS. The possible metabolic process could be as follows: G-Rb1 (or PDS)-->G-Rd-->G-F2-->CK-->Ppd. PMID- 12579939 TI - [Analysis and structure identification of trace constituent in the total ginkgolide by using LC/DAD/ESI/MS]. AB - AIM: To identify the trace constituents in the total ginkgolide from leaves of Ginkgo biloba. METHODS: The extract was analyzed by using LC/DAD/ESI/MS combination technology, both UV and mass spectrums of each compound were recorded simultaneously. UV, IR, MS and NMR spectra were employed for structural identification. RESULTS: With higher sensitivity of the MS detector, three trace constituents were found as well as the known compounds ginkgolide A (GA), ginkgolide B (GB), ginkgolide C (GC) and bilobalide (BD). CONCLUSION: Two compounds (tR 57.8 and 56.7 min) were identified as 1, 10-dihydroxy-3, 14 didehydroginkgolide and 10-hydroxy-3, 14-didehydroginkgolide, named as ginkgolide K and ginkgolide L, respectively, according to the characteristics of their UV, MS, IR and NMR spectra. It is the first time of finding the ginkgolide compounds containing double bond. PMID- 12579940 TI - [Separation and quantitative analysis of ginkgolic acids from Ginkgo biloba leaves by reverse phase argentation high performance liquid chromatography]. AB - AIM: To develop a reverse phase argentation high performance liquid chromatographic (RP-AHPLC) method for the separation and determination of ginkgolic acids. METHODS: Liquid chromatography-electrospray ionization mass spectrometry (LC/ESI-MS) was applied to identify ginkgolic acids from Ginkgo biloba leaves and four ginkgolic acids of the samples were separated and quantified by RP-AHPLC. Leaves were extracted with ethanol and analytes were extracted with hexane after addition of acid/salt solution and adsorbent to matrix solution. Ginkgolic acids were separated and determined within 30 minutes by RP-AHPLC under optimum chromatographic conditions. Methanol and 5% aqueous acetic acid (90:10) containing 0.03 mol.L-1 silver ion was used as mobile phase, column temperature was selected at 30 degrees C, flow rate was 1.0 mL.min-1, UV detection wavelength was at 310 nm. The spectra analysis and purity identification of chromatographic peaks of ginkgolic acids were further confirmed by means of diode array detection. RESULTS: Four ginkgolic acids were baseline separated from each other and from other interfering components. The average recovery and relative standard deviation of the method were 97.3% and 1.6%, respectively. CONCLUSION: RP-AHPLC was an excellent method for separation of homologous with different carbon atom numbers and double bond. The method is useful for the quality control of extract of Ginkgo biloba leaves. PMID- 12579941 TI - [Terbium sensitized chemiluminescence of enoxacin]. AB - AIM: To determine the enoxacin by means of terbium sensitized chemiluminescence in pharmaceutical preparations as well as urine samples. METHODS: Through six-way injection valve Ce4+, H2SO3, Tb3+ and enoxacin (ENX) standard solutions were injected into the flow system in a certain order, the chemiluminescence signal was detected by weak luminescence analyzer. RESULTS: The chemiluminescence intensity was linearly related to the concentration of ENX in the range of 2.0 x 10(-9)-6.0 x 10(-8) mol.L-1 with a detection limit of 4.0 x 10(-10) mol.L-1. Relative standard deviation was less than 1.6% (n = 11). CONCLUSION: It provides a sensitive rapid, simple and accuracy measurement of enoxacin for the pharmacokinetic studies. PMID- 12579942 TI - [Studies on the pharmacokinetics and relative bioavailability of salbutamol aerosol in healthy volunteers]. AB - AIM: To study the pharmacokinetics and relative bioavailability of salbutamol aerosol in healthy volunteers. METHODS: An HPLC method for the determination of salbutamol in human plasma was improved. Ten healthy male subjects were enrolled. A randomized, two-way crossover, open design was adopted. After the subjects inhaled or orally administered salbutamol, fourteen blood samples were taken at predetermined time. The concentrations of salbutamol in plasma were determined by HPLC, and then assessed with PCNONLIN software to obtain the pharmacokinetic parameters and relative bioavailability of aerosol versus water solution. RESULTS: The standard curve was linear over the range 0.2-20 ng.mL-1. The intra- and interassay RSDs were 7.01% and 2.10% at 0.4 ng.mL-1, 2.18% and 5.25% at 4.0 ng.mL-1 and 4.61% and 4.85% at 15.0 ng.mL-1. The recoveries were between 90% and 110%. The pharmacokinetics of salbutamol aersol was described well with a two compartment model, and the parameters for salbutamol inhaled and orally administered were assessed as follows: Tmax were (0.22 +/- 0.07) h and (1.8 +/- 0.6) h, Cmax were (3.4 +/- 1.1) ng.mL-1 and (3.9 +/- 1.4) ng.mL-1, T1/2 beta were (4.5 +/- 1.5) h and (4.6 +/- 1.1) h, respectively. The AUC0-20 min (inhal) was 7.94 times as high as the AUC0-20 min (p.o.). There were significant differences between Tmax, AUC, K12, K21, alpha and T1/2 alpha (P < 0.05). The relative bioavailability of salbutamol aerosol was 57.23% compared with its oral solution. CONCLUSION: The assay was sensitive, specific, accurate and precise. The absorption process of salbutamol aerosol in human was significantly different from that of the oral solution. It was demonstrated that AUC0-20 min reflected the lung availability of salbutamol inhaler. PMID- 12579943 TI - [Permeation of buspirone hydrochloride across animal buccal mucosa and its mechanism]. AB - AIM: To study the permeation of busprione hydrochloride across animal buccal mucosa and its permeation mechanism. METHODS: The in vitro penetration model was designed to investigate drug buccal mucosa permeability, and the influence factors (different animal oral mucosa, such as rabbit, guinea pig, pig, sheep and cow, drug concentrations and solution pH values) were measured. RESULTS: The permeability of rabbit buccal mucosa for busprione hydrochloride was the largest. The permeation rate was increased with solution pH and directly to the drug concentration. CONCLUSION: The transport way of busprione hydrochloride was passive diffusion and the transport channel was through cells. PMID- 12579944 TI - [Study on the enzyme dependant colon targeting prodrug--dexamethasone succinate dextran and its tablets]. AB - AIM: To develop an orally administered colon targeting drug delivery system dexamethasone succinate dextran (DSD) tablets. METHODS: Dexamethasone succinate dextran was synthesized in an anhydrous environment. Using 4-dimethyl aminopyridine and 1,1'-carbonyldiimidazole as the catalyzer. The chemical structure was identified by UV, IR, NMR and MS. The contents of dexamethasone in various samples were determined by HPLC. RESULTS: Dexamethasone was distributed mainly in plasma and gastric contents after the oral administration of common tablets. In contrast, after oral administration of DSD tablets, the recovery of dexamethasone in plasma and gastric contents decreased significantly, while the percentage of dexamethasone in cecum and colon increased obviously. CONCLUSION: The experimental results showed the good colon targeting property of DSD prodrug compared with free dexamethasone. PMID- 12579946 TI - [Advances of using the nasal route for drug delivery to the brain]. PMID- 12579945 TI - [Study on allele-specific diagnostic PCR of the traditional Chinese medicines of the deers]. AB - AIM: To develop a convenient and accurate method of DNA molecular marker for the identification of traditional Chinese medicines made of deers, consisting of pilose antler, penis and testis, tendon and foetus. METHODS: Based on the analysis of DNA sequence of mitochondrial Cyt b gene from original animals of both genuine crude drugs, Cervus nippon and Cervus elaphus, and adulterants, a pair of allele-specific primers named as ILu01-L and ILu01-H were designed for distinguishing geniune crude drugs of deers from their adulterants. RESULTS: The results of diagnostic PCR annealing at 64 degrees C for original animals showed that a 365 bp fragment was only amplified from DNA templates of Cervus nippon and Cervus elaphus. For the identification of medicinal materials total of 43 samples from 6 packages were tested under the same reaction conditions except for DNA templates extracted from these crude drugs. Only 9 samples mentioned above was shown to generate positive amplificon. The result indicate that of 8 samples from 1 package of pilose antler and only 1 sample of deer tendon was genuine crude drug. After that, 3 amplified fragments selected randomly were performed with sequencing analysis with the purpose of verifying the results from diagnostic PCR. Data from sequencing confirmed the reliability of diagnostic PCR identification. CONCLUSION: The diagnostic primers designed in the present study were highly specific for Cervus nippon and Cervus elaphus, and they could be used for the authentication of traditional Chinese medicines made from the deer. The quality of the crude drugs of the deer in the current market is a problem and more effective quality control for these traditional Chinese medicines is urgently needed. PMID- 12579947 TI - [Inhibition of human low-density lipoprotein oxidation by salvianolic acid-A]. AB - AIM: Oxidized low-density lipoprotein (LDL) is involved in the development of atherosclerosis. Oxidative modulation of serum LDL is related to oxygen free radicals. Antioxidants have beneficial effects on oxidative modulation of LDL and development of atherosclerosis. Salvia miltriorhiza (Danshen) preparations have been widely used in the treatment of cardio-cerebral vascular diseases in China. Salvianolic acid A (Sal-A), one of the components of Salvia miltriorhiza, was shown to have strong antioxidative activity. The aim of this investigation was to evaluate the effect of Sal-A on human LDL oxidative modulation mediated by copper ions. METHODS: Oxidation of human LDL was performed in pH 7.4 phosphate-buffered saline with 10 mumol.L-1 CuSO4 at 37 degrees C water for 20 h. The content of malondialdehyde (MDA), lipofuscin and vitamin E in LDL as well as the rate of electrophoretic mobility (REM) of LDL were measured. The generation of free radicals during LDL oxidation was detected by low level-chemiluminescence (LL CL). The chelation of Cu2+ by Sal-A was detected by UV-spectrum scanning. RESULTS: Sal-A (10(-6) to 10(-4) mol.L-1) was shown to markedly reduce the production of MDA and lipofuscin as well as the consumption of vitamin E during LDL oxidation. Sal-A (10(-4) mol.L-1) was also shown to inhibit the increase of REM of LDL caused by oxidative modification. In addition, the spectrum of LL-CL showed that Sal-A (10(-6) to 10(-5) mol.L-1) decreased the generation of free radicals during LDL oxidation in a dose dependent manner. The differential UV spectrum of Sal-A in the presence of Cu2+ indicated that Sal-A could chelate copper ions. CONCLUSION: Sal-A has inhibitory effect on Cu2+ mediated human LDL oxidation through chelating Cu2+ and scavenging free radicals. PMID- 12579948 TI - [Expression of vascular endothelial growth factor in U937 foam cells and the inhibitory effect of drugs]. AB - AIM: To study the expression of vascular endothelial growth factor (VEGF) in U937 foam cells and the inhibitory effect of salvianolic acid B and Ginkgo biloba extract in vitro. METHODS: U937 cells were incubated with 80 mg.L-1 oxidized low density lipoprotein (OX-LDL) for 48 h and a macrophage-derived foam cell model was established. The VEGF concentration in the media was determined by ELISA; the VEGF protein expression in cells was measured with immunohistochemistry; the VEGF mRNA level in cells was measured by in situ hybridization; the positive ratio detected by a morphometrical analysis system was used as the amount of the VEGF protein expression and the mRNA level. RESULTS: After U937 cells were incubated with OX-LDL, VEGF expression level increased greatly both in the cells and in the media. Salvianolic acid B and Ginkgo biloba extract were shown to remarkably inhibit the increase of VEGF. After treated with 10 micrograms/L-1 salvianolic acid B and Ginkgo biloba extract, the VEGF protein concentration in the media and positive ratio in the cells decreased compared with foam cells. After treated with 10 micrograms.L-1 salvianolic acid B and 100 micrograms.L-1 Ginkgo biloba extract, the VEGF mRNA level decreased measured by in situ hybridization. CONCLUSION: A high VEGF expression level was determined in U937 foam cells. Salvianolic acid B and Ginkgo biloba extract were found to inhibit VEGF expression significantly in U937 foam cells in vitro. PMID- 12579949 TI - [Inhibitory effects of ginkgolide B on proliferation of bovine aortic smooth muscle cells]. AB - AIM: To investigate the effects of ginkgolide B (GB) on proliferation of bovine aortic smooth muscle cells (SMC) and its related mechanisms. METHODS: After pretreating with GB or the mixture of ginkgolide A and B (GA:GB) at 37 degrees C for 0.5 h, the VSMC were treated with or without angiotensin II (Ang II) for 24 h. The proliferation of SMC was evaluated by 3H-thymidine incorporation and the cell cycle phase was measured by flow cytometry. Cytotoxicity was reflected by MTT and lactate dehydrogenase (LDH) activity of the supernatant. RESULTS: Whether or not treated with Ang II, GB and GA:GB were shown to suppress SMC proliferation in concentration-dependent fashion at concentrations ranging from 10(-9) mol.L-1 to 10(-5) mol.L-1. The inhibitory effects appeared to be related to a G1-->S block in cell cycle traverse. CONCLUSION: The suppression of SMC proliferation by GB might not only be contributed by blockage of the PAF receptor activity. PMID- 12579950 TI - [1 alpha, 25-dihydroxyvitamin D3 and its analogues modulate the phagocytosis of human monocyte-derived dendritic cells]. AB - AIM: To investigate the role of 1 alpha, 25-dihydroxyvitamin D3 (calcitriol) and its analogues tacalcitol and 24, 25(OH)2D3 on the phagocytosis of human monocyte derived dendritic cells (MoDC). METHODS: MoDC were generated in vitro by differentiating monocytes in the presence of GM-CSF and IL-4 for 5 days. Expression of mannose receptor (MR) and Fc gamma receptors (Fc gamma Rs) by MoDC was analysed by flow cytometry. Zymosan ingestion was measured to assess the phagocytosis of MoDC. RESULTS: MoDC expressed high level of MR and Fc gamma Rs and showed the capacity of zymosan ingestion. Calcitriol and tacalcitol but no 24, 25(OH)2D3 not only upregulated the expression of MR and Fc gamma Rs on MoDC but also correspondingly enhanced their phagocytosis by increasing zymoasan ingestion. Furthermore, the upregulatory role occurred in the early stage of MoDC differentiation and was irreversible. The upregulatory role of calcitriol was dose dependent. CONCLUSION: Calcitriol and its analogue tacalcitol may play an important role in dendritic cell binding and capturing foreign antigens at the initiation of immune response. PMID- 12579951 TI - [Protective effects of tacrine and donepezil against staurosporine-induced apoptotic death]. AB - AIM: To study whether tacrine and donepezil can prevent cell apoptosis induced by staurosporine in NG108-15 and Hela cell lines. METHODS: MTT assay was used to examine if staurosporine impairs cell metabolism. Phase-contrast and fluorescence microscope were used to examine cell morphological changes. DNA was isolated and electrophoretically separated on 1% agarose gel to observe if there were DNA fragments. Western blot was made to analyse protein levels of anti-apoptotic Bcl 2 and proapoptotic Bax. RESULTS: NG108-15 cells treated with 0.1 mumol.L-1 staurosporine for 12-24 hours exhibited marked cell death and DNA fragmentation. Pre-treatment with 0.1 mmol.L-1 tacrine provided approximately 40% protective effect and resulted in obvious inhibition or delay of DNA fragmentation. Moreover, NG108-15 cells treated with tacrine became elongated and polarized, and showed longer processes than control cells. Pretreatment with 0.1 mmol.L-1 tacrine significantly increased the expression of Bcl-2 protein level and delayed the staurosporine-induced increase of Bax protein expression. However, donepezil did not show any protective effect on the cell impairment induced by staurosporine in NG108-15 cells. In Hela cells 0.1 mumol.L-1 staurosporine also induced significant cell injury, but pretreatment with tacrine and donepezil did not provide any obvious protective effect against this cell damage. CONCLUSION: Donepezil did not provide obvious protective effect against apoptosis, and protective effects of tacrine might not be mediated through AChE inhibition. Protective effects of tacrine against staurosporine-induced injury might be selective to different cells. PMID- 12579952 TI - Inhibitory effect of meloxicam on human polymorphonuclear leukocyte adhesion to human synovial cell. AB - AIM: To investigate the effect of meloxicam on human polymorphonuclear leukocyte (PMN) adhesion to human synovial cell (HSC), and to explore its mechanism. METHODS: MTT colorimetry was used to determine the adhesion effect of PMN to HSC. Cell-ELISA and RT-PCR methods were used to determine the expression of ICAM-1 and VCAM-1. Nuclear transcription factor-kappa B (NF-kappa B) was measured by electrophoretic mobility shift assay (EMSA) method. RESULTS: Meloxicam was found to effectively inhibit TNF-alpha (50 u.mL-1 for 12 h) and IL-1 beta (50 u.mL-1 for 12 h)-induced adhesion of PMN to HSC (IC50 3.38 x 10(-7) mol.L-1 and 3.56 x 10(-6) mol.L-1, respectively) in a concentration-dependent manner. ICAM-1 protein and mRNA expression induced by TNF-alpha (50 u.mL-1) were inhibited by meloxicam at 1 x 10(-6)-1 x 10(-5) mol.L-1. The activation of NF-kappa B was also inhibited by meloxicam at 1 x 10(-6)-1 x 10(-5) mol.L-1. CONCLUSION: These results suggest that meloxicam inhibit TNF-alpha stimulated PMN-HSC adhesion and expression of ICAM-1 by suppressing the activity of NF-kappa B. PMID- 12579953 TI - Experimental treatment of complications in alloxan diabetic rats with alpha glucosidase inhibitor from the Chinese medicinal herb ramulus mori. AB - AIM: To assess the effects of the alpha-glucosidase inhibitor Sangzhi (Ramulus mori, SZ) on the relief of diabetic symptoms of hyperglycemia and the prevention of its late complications in alloxan diabetic rats with high-calorie chow. METHODS: The aqueous extract of Sangzhi was given orally to alloxan diabetic rats for 15 days. The hyperglycemic symptoms were observed. The blood glucose, lipid levels and the nephrotic representations were measured. RESULTS: When alloxan diabetic rats on high-calorie chow were treated with SZ, the hyperglycemic symptoms were improved, the blood lipid levels were improved, the ratio of kidney over body weight and the blood N-acetyl-beta-D-glucosaminidase (NAG) activity were lowered. The degree of renal pathological changes was significantly reduced. CONCLUSION: SZ may be useful for treating diabetes and its complications. PMID- 12579954 TI - [Synthesis of 4-, 6- or 7-substituted phenyliminomethylenecoumarins and their anticancer activities]. AB - AIM: A series of substituted phenyliminomethylenecoumarins derivatives was designed in order to find compounds possessing anticancer activities. METHODS: Title compounds (1a-b, 2a-b and 3a-q) were synthesized and screened by several anticancer models in vitro. RESULTS: Twenty-one new compounds (1a-b, 2a-b and 3a q) were synthesized and screened. Structures of the new compunds were determined by MS, HNMR and elemental analysis. Twelve compounds (3c, 3d, 3e, 3f, 3g, 3h, 3j, 3k, 3m, 3o, 3p, 3q) showed inhibitory effects on HCT-8, KB and Bel7402 cell lines in vitro. CONCLUSION: Some compounds had certain anticancer activities and were worth further studying. PMID- 12579955 TI - [Studies on the triterpenoid constituents from the aerial part of Cimicifuga foetida L]. AB - AIM: To look for new active constituents from the aerial part of Cimicifuga foetida L. METHODS: Various column chromatographic techniques were used for the isolation and purification of the ingredients. The structure were elucidated on the basis of spectral evidences and chemical reaction. RESULTS: Five compounds were obtained and identified as 23-O-acetylshengmanol-3-O-alpha-L arabinopyranoside (1), 23-O-acetylshengmanol-3-O-beta-D-xylopyranoside (2), 25 anhydrocimigenol-3-O-beta-D-xylopyranoside (3), cimigenol-3-O-alpha-L arabinopyranoside (4), cimigenol-3-O-beta-D-xylopyranoside (5). CONCLUSION: Compound 1 is a new compound, and compounds 2 and 4 were isolated from this plant for the first time. PMID- 12579956 TI - [Studies on water-soluble constituents of Echinacea prupurea]. AB - AIM: To study the water-soluble constituents of Echinacea purpurea. METHODS: The compounds were isolated by chromatography method and their structures were identified on the basis of spectral analyses. RESULTS: Five compounds were identified as 2-(4-hydroxylphenyl)-ethyl-O-alpha-L-rhamnopy-ranoasyl-(1-->6)-beta D- glucopyranoside, named echipuroside A 1); (6S, 9R)-6-hydroxy-3-one-alpha inonol-9-O-beta-D-glucopyranoside 2), ampelopsisionoside 3); phenylmethyl-6-O beta-D-xylopyranosyl(1-->6)-beta-D-glucopyranoside 4) and phyenylmethyl-beta-D glucopyranoside 5). CONCLUSION: Compound 1 is a new compound, the others were isolated from this plant for the first time. PMID- 12579957 TI - [Studies on chemical constituents of Uvaria macrophylia]. AB - AIM: To study the chemical constituents from Uvaria macrophylla Roxb. (Annonaceae). METHODS: Various chromatography techniques were used to separate and purify the constituents. Their structures were elucidated by UV, IR, MS, 1HNMR, 13CNMR, 1H-1H COSY, HMQC and HMBC spectral analysis. RESULTS: Seven compounds have been isolated from the CHCl3 extract of the roots of the U. macrophylla. They were identified as macrophyllin (1), onysilin (2), taraxerol (3), 3,5-dimethoxy benzyl benzoic acid ester (4), benzoic acid (5), beta sitosterol (6) and daucosterol (7). CONCLUSION: Compound 1 is a new compound. Compounds 2-7 were obtained from this plant for the first time. PMID- 12579958 TI - [Isolation and structure of wilfordlonging from Tripterygium wilfordii Hook. f]. AB - AIM: To study the chemical composition of Tripterygirm wilfordii Hook. f. METHODS: Colunm chromatography was used to separate the chemical constituents. UV, IR, MS, HRMS, 1HNMR, 13CNMR (COM and OFR), 1H-1H COSY, 1H-13C COSY, NOESY and COLOC spectra were used to determine the structures of the isolated constituents. RESULTS: Two sesquiterpene alkaloids were isolated and their structures were elucidated as wilforgine and wilfordlongine on the basis of spectral evidence. CONCLUSION: Wilfordclonine is a new sesquiterpene alkaloid. PMID- 12579959 TI - [HPLC determination of insulin and its related substances in insulin powder for inhalation]. AB - AIM: To determine insulin and its related substances in insulin powder for inhalation by reversed phase high performance liquid chromatography method. METHODS: The initial mobile phase was solution A (0.1% trifluoroacetic acid acetonitril 70:30) and changed to solution B (0.1% trifluoroacetic acid acetonitril 60:40) in 30 minutes. The flow rate was 2.0 mL.min-1, the column temperature was 30 degrees C, the wave length was 280 nm, the injected volume was 20 microL. RESULTS: Insulin was well separated from other peaks induced in different conditions. There was good linear relationship between the amount of insulin and its peak area, the RSD was 1.1%, the insulin solution for determination was stable in 12 hours, and the quantity detected was near the added. CONCLUSION: The method is simple and accurate to detect insulin and its related substances in insulin and its preparations. PMID- 12579960 TI - [Determination of ofloxacin in human fallopian tube, uterus and serum by high performance liquid chromatography]. AB - AIM: To establish a method for determineation of the concentration of ofloxacin in human fallopian tube, uterus and serum. METHODS: The separation was performed on a Spherisob C18 column (Hypersil, 250 mm x 4.6 mm ID, 5 microns) with a mobile phase of acetonitrile-0.01 moL.L-1 potassium dihydrogen phosphate-0.5 mol.L-1 tetrabutylammonium bromide (9:91:4, pH 2.5). The flow rate was 1.0 mL.min-1 and detection was at 294 nm. The samples were homogenated or ground to powder after freezing with liquid nitrogen. 1% triton-100 and certain volume of ethylacetate isopropanol (10:1) were added, shaken and centrifuged. Then the entire organic layer was transferred to a tube and vacuum dried. The residue was reconstituted in the mobile phase for HPLC. RESULTS: There was a linear relationship between the peak area ratio and the ofloxacin concentration over the range of 0.2-8.0 micrograms.mL-1. The limits of detection was 40 ng.mL-1. Using this method to determine the ofloxacin concentrations in relevant organs as well as in the plasma of patients of the Department of Gynecology, and achieved satisfactary results. CONCLUSION: The method can be applied to assay the ofloxacin concentration in human tissues. Ofloxacin was well distributed in woman fallopian tube, uterus and serum after single oral administration. PMID- 12579961 TI - [Identification of estazolam, alprazolam and triazolam in human urine by LC/MSn]. AB - AIM: To investigate the fragmentation behavior of triazolobenzodiazepines and to develop a specific, sensitive and rapid LC/MSn assay for simultaneous determination of estazolam, alprazolam and triazolam in human urine. METHODS: After oral administration of a single 4 mg dose of the drugs to each of three healthy volunteers, urine samples were purified by solid-phase extraction, and then injected into an ODS column (150 mm x 4.6 mm) with a mobile phase of methanol-water (8:2) for LC/MSn analysis. The structures of estazolam, alprazolam and triazolam in human urine were identified by direct comparison of the observed mass spectra and the chromatographic retention time with those of the reference substance. The mass spectrometer (Finnigan LCQ) was operated in positive mode and in two scan modes including SIM and full scan MS/MS mode. The obtained mass spectra was analyzed assisted with the software Mass Frontier 1.0 for their fragmentation pathways. RESULTS: The full scan MS/MS spectra of each compound gave characteristic fragment ions of [M + H - N2]+ and [M + H - Cl]+. The detection limit was below 0.5 ng.mL-1 for estazolam, alprazolam and triazolam in human urine. CONCLUSION: The method is useful in forensic and clinical toxicology in which unequivocal identification of eatazolam, alprazolam and triazolam is desired. PMID- 12579962 TI - [Isolation and identification of a major metabolite of SFZ-47 in the rabbit urine]. AB - AIM: To isolate and identify a glucuronide metabolite of SFZ-47 [3H-1,2-dihydro-2 (4-methyl-phenylamino)methyl-1-pyrrolizinone], which is difficult to synthesize because it undergoes hydrolysis and intramolecular acyl migration at physiological pH, in rabbit urine. METHODS: Two rabbits were ig 200 mg doses of SFZ-47. Urine was collected for 24 h, adjusted to pH 4.0 with acetic acid and lyophilized. The residues were reconstituted in 25 mL methanol and centrifuged at 5,000 r.min-1 for 10 min. The supernatant was filtered (0.45 micron) and then isolated with semi-preparative reversed phase HPLC. The eluent collected from individual peaks was evaporated by rotary evaporation and freeze-drying. Compounds were then identified with electrospray ion trap mass spectrometry and 1HNMR spectroscopy. RESULTS: The 1HNMR and ESI-MSn results indicate that the metabolite is the 1-O-acyl beta-D-glucuronide conjugate of 4-(3H-1,2-dihydro-1 pyrrolizinone-2-methylamino) benzoic acid. CONCLUSION: This method was shown to be rapid and simple and gave excellent resolution from endogenous constituents in urine, and it is suitable for preparation of the glucuronide metabolites of SFZ 47 and its analogues. PMID- 12579963 TI - [Study on water of crystallization in tazobactam]. AB - AIM: To investigate whether tazobctam has crystallization water. METHODS: In order to establish a method for water content determination in tazobctam, many methods including Karl Fischer titration, weight loss on drying, TGA, DSC and X ray diffraction experiment of an orthorhombic form were studied. RESULTS: The water content of the same batch tazobactam determined by different mthods was different. CONCLUSION: The results showed that one mole tazobatam contains half mole water of crystallization and the strength of attraction between tazobatam molecule and water molecule is rather strong. PMID- 12579964 TI - [Effect of light and heat on the stability of furacilin aqueous solution]. AB - AIM: To study the effect of both light and heat on the stability of furacilin aqueous solution and the probability of substituting for isothermal accelerated tests by nonisothermal accelerated tests upon exposure to light at high temperatures. METHODS: The isothermal and nonisothermal accelerated tests were employed. The accelerated tests were proceeded in the dark and exposed to light at high temperature. Tungsten, ultraviolet and fluorescent lamps were employed in exposure tests. RESULTS: The degradation of furacilin aqueous solution in isothermal heating experiments or the exposure experiments to light at high temperatures obeys zero-order kinetics. The total degradation rate constant k caused by both light and heat can be divided into two parts: k = kdark + klight, where kdark and klight are the degradation rate constant caused by heat and light, respectively. The klight can be expressed as klight = Alight.exp( Ea,light/RT).E, where E is the illuminance of light; Alight and Ea,light are both experimental constants. The parameters obtained in nonisothermal accelerated tests were comparable to those obtained in classic isothermal accelerated tests. CONCLUSION: Nonisothermal accelerated tests may substitute for isothermal accelerated tests during the study of the effects of both light and heat on the stability of drugs, in order to save time, labor and drugs. PMID- 12579965 TI - [Optimization of hybridization efficiency in cDNA chip technology]. AB - AIM: There are many factors affecting hybridization in cDNA chip: DNA concentration immobilized on glass surface, spotting solution dissolving DNA, the concentration and purity of fluorescence-labeled probe, hybridization solution, hybridization temperature, hybridization time, Cy3- and Cy5-labeled probes, etc. In order to improve hybridization efficiency, tests were designed to optimize these factors. METHODS: Factors are changed one by one. Optimal values are selected by comparing those of different groups. RESULTS AND CONCLUSION: The efficient hybridization condition is as follows: 0.50 microgram.microL-1 DNA fragments resolved in 1 x Micro Spotting Solution (ArrayIt) hybridize with purified fluorescent probe in 0.5% SDS/10 x SSC at 42 degrees C. PMID- 12579966 TI - [NMR-based screening of protein inhibitors in drug discovery]. PMID- 12579967 TI - [Effect of bicyclol on acetaminophen-induced hepatotoxicity: energetic metabolism and mitochondrial injury in acetaminophen-intoxicated mice]. AB - AIM: To investigate the mechanism of the protective effect of bicyclol on acetaminophen-induced hepatotoxicity in mice. METHODS: 31P-MRS spectra in vivo were determined by using surface coil technique. The membrane fluidity of mitochondria and the activity of mitochondrial ATPase were also determined by spectrofluorophogometry and spectrophotometry methods. RESULTS: The hepatotoxicity of acetaminophen is related to the lipid peroxidation and covalent binding to macromolecules, which leads to damage of mitochondrial function. Our results showed that the decrease of ATP/Pi and the elevation of PME/ATP in acetaminophen-intoxicated mice were significantly inhibited by two doses of bicyclol (100, 200 mg.kg-1) pretreatment, which indicate that bicyclol has significant protective effect on the decrease of liver ATP content induced by acetaminophen. Acetaminophen significantly inhibited the activity of mitochondrial ATPase by its cytotoxic metabolite NAPQI [N-acetyl-p-benzoquinone], which has the potential to react with sulfhydryl groups or through sulfhydryl group oxidation. Our results showed that the reduction of mitochondrial fluidity as well as the inhibitory effect of mitochondrial ATPase induced by acetaminophen were also reduced by bicyclol. CONCLUSION: The effect of bicyclol on acetaminophen-induced liver injury maybe partly due to its protective effects on hepatic energy metabolism and mitochondria function. PMID- 12579968 TI - [Effects of leflunomide on immunological liver injury]. AB - AIM: To study the effect of leflunomide (Lef) on immunological liver injury in mice. METHODS: Immunological liver injury was induced by tail vein injection of BCG + LPS (BCG 2.5 mg, 1 d, LPS 10 micrograms, 10 d). The ALT, AST, NO level in plasma and MDA, GSHpx in liver homogenate were assayed by spectroscopy. The serum content of TNF-alpha was determined by ELISA. IL-1, IL-2 and ConA-induced splenocyte proliferation response were determined by methods of 3H-infiltrated cell proliferation. RESULTS: Immunological liver injury induced by BCG + LPS was successfully duplicated. Lef (4, 12, 36 mg.kg-1) was found to significantly decrease the serum transaminase (ALT, AST) activity and MDA content in liver homogenate, and improved reduced GSHpx level of liver homogenate. Furthermore, Lef (4, 12, 36 mg.kg-1) significantly reduced TNF-alpha and NO level in serum, and IL-1 production by PM psi. Moreover, the decreased IL-2 production and ConA induced splenocyte proliferation response were further inhibited. CONCLUSION: Lef showed significant protective action on immunological liver injury in mice. PMID- 12579969 TI - [Effects of intracerebroventricular administration of endomorphin-1 on blood pressure in anesthetized rats]. AB - AIM: To observe the effects of intracerebroventricular administration of endomorphin-1 on blood pressure in anesthetized rats and to assess its mechanism. METHODS: Variations of mean arterial pressure (MAP) were observed after icv injection of endomorphine-1 in rats. The effects of i.v. or icv of various blockers, naloxone, phentolamine, propranolol, atropine and N omega-nitro-L arginine, on the variation of MAP caused by endomorphin-1 were observed. RESULTS: Endomorphin-1 was shown to decrease MAP. The decrease in MAP was blocked by naloxone. Pretreatments with icv phentolamine and propranolol showed no effect on the vasodepression induced by icv endomorphin-1. However, pretreatment with icv atropine (25 micrograms.kg-1) attenuated the vasodepression. Pretreatments with i.v. phentolamine, propranolol and N omega-nitro-L-arginine showed no effect on the vasodepression induced by icv endomorphin-1. But, pretreatment with i.v. atropine (50 micrograms.kg-1) and bilateral vagotomy attenuated the vasodepression. CONCLUSION: Intracerebroventricular administration of endomorphin 1 produces vasodepressor response in anesthetized rats, which is mediated by opioid receptor. The vasodepression is associated with the central M cholinoceptor and the excitation of the vagus. PMID- 12579970 TI - [Inhibitory effects of benzoisoselenothiazolidone sulfonamide derivatives on cyclooxygenase]. AB - AIM: To study the inhibitory effects of benzisoselenothiazolidone sulfonamide derivatives on cyclooxygenase. METHODS: 6-Keto-PGF1 alpha and PGE2 were assayed by radioimmunoassay (RIA) method; mRNA expression of COX-1 and COX-2 were assayed by reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: Compound A [N-4-(4-methoxyphenyl-aminosulfonyl)-benziososelenothiazolidone] and B [N-4-(4-fluorophenyl-aminosulfonyl)-benzoisoselenothiazolidone] were two benzoisoselenothiazolidone sulfonamide derivatives, which can inhibit COX activity with IC50 of 1.5 x 10(-8) mol.L-1 and 5.0 x 10(-8) mol.L-1 for COX-2, as well as IC50 of 1.5 x 10(-5) mol.L-1 and 2.8 x 10(-5) mol.L-1 for COX-1. The ratio of IC50 COX-1/IC50 COX-2 of compound A and B are 1,000 and 560, respectively. They both can inhibit COX-2 mRNA expression in cultured rat peritoneal macrophages stimulated with LPS (1 microgram.mL-1), and have no effect on COX-1 mRNA expressions. CONCLUSION: Compound A and B, two benzoisoselenothiazolidone sulfonamide derivatives, both are selective inhibitory agents of COX-2, and possess inhibitory effects on 5-lipoxygenase and cyclooxygenase. PMID- 12579971 TI - [Synthesis and determination for enantiomeric purity of 6-fluoro-L-DOPA]. AB - AIM: To study the synthesis and determination for enantiomeric purity of 6-fluoro L-3, 4-dihydroxyphenylalanine (6-fluoro-L-DOPA, 6-FDOPA). METHODS: 2-(2-Fluoro-4, 5-dimethoxybenzyl)-N-(diphenylmethylene) glycine tert-butyl ester (8), a new compound, was synthesized from the starting material nitroveratraldehyde via the nucleophilic displacement, reductive iodination, and chiral catalytic phase transfer alkylation, and 6-FDOPA was prepared from hydrolysis of 8. The enantiomeric purity of 6-FDOPA was determined by HPLC method using a chiral mobil phase and reversed-phase C18 column. RESULTS: The total time of synthesis was less than 90 min, the overall chemical yield from potassium fluoride was about 33%, and the enantiomeric purity was above 95%. CONCLUSION: Large scale production of 6-FDOPA and automatic synthesis of 6-[18F]fluoro-L-DOPA with excellent chemical and entiomeric purity are available. The practical technique was provided for the radiochemical synthesis and entiomeric purity of 6 [18F]fluoro-L-DOPA. PMID- 12579972 TI - [Studies on the chemical constituents of Evodia rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang]. AB - AIM: To study the chemical constituents of the unriped fruits from Evodia rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang. METHODS: Various chromatographic techniques were used to separate and purify the constituents. Their structures were elucidated on the physico-chemical properties and spectral data. RESULTS: A lot of compounds were isolated of the unriped fruits from Evodia rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang. This paper reports mainly compounds identified as shihulimonin A, emodin, physcion, chrysophanol and limonin. CONCLUSION: Shihulimonin A is a new compound. Emodin, physcion and chrysophanol were isolated from the Evodia for the first time. PMID- 12579973 TI - [Study on chemical constituents of Hemistepta lyrata Bunge]. AB - AIM: To separate and identify the chemical constituents of the whole plant of Hemistepta lyrata Bunge. METHODS: Compounds wese separated with chromatography and their chemical strucrures were elucidated with UV, IR, EIMS, FABMS, ESIMS, 1HNMR, 13CNMR HMQC and HMBC spectral methods. RESULTS: Five compounds were isolated from the plant. They were identified as apigenin I, apigenin-7-O-beta-D lutinoside II, acacetin-7-O-beta-D-lutinoside III, astragalin IV, hemislienoside V. CONCLUSION: These compounds were isolated from this plant for the first time, and compound V is a new one. PMID- 12579974 TI - [Studies on the chemical constituents from Tribulus terrestris]. AB - AIM: To investigate the chemical constituents of the fruit of Tribulus terrestris J.. METHODS: Various chromatographic techniques were used to separate the chemical constituents. ESIMS, IR, 1HNMR, 13CNMR and HMBC were used to determine the structures of the isolated constituents. RESULTS: Two new compounds were isolated from the fruits of Tribulus terrestris J. and were identified as neohecogenin-3-O-beta-D-glucopyranosyl (1-->2)-beta-D-glucopyranosyl (1-->4)-beta D-galactopyranoside (I); neohecogenin-3-O-beta-D-glucopyranosyl (1-->4)-beta-D galactopyranoside (II). CONCLUSION: Compounds I and II are new steroidal saponins. PMID- 12579975 TI - [Chemical constituents from the leaves of crataegus pinnatifida Bge. var. major N.E.Br]. AB - AIM: To study the chemical constituents in the leaves of Crataegus pinnatifida Bge. var. major N.E.Br.. METHODS: Compounds were isolated by using porous resin, silica gel, polyamide chromatographic techniques etc. Their structures were elucidated by chemical and spectroscopic methods. RESULTS: Eight compounds were identified as pinnatifin I (1), quercetin (2), 3-O-beta-D-glucopyranosyl quercetin (3), 3-O-beta-D-galacopyranosyl quercetin (4), 3-O-beta-D glucopyranosyl (6-->1)-alpha-L-rhamnosyl quercetin (5), 3-O-beta-D galacopyranosyl (6-->1)-alpha-L-rhamnosyl quercetin (6), kaempferol (7), 7-O alpha-L-rhamnosyl-3-O-beta-D- glucopyranosyl kaempferol (8). CONCLUSION: Compound 1 is a new compound. Compound 8 was isolated from this plant for the first time. PMID- 12579976 TI - [LC-MS analysis of related substances in ampicillin and amoxicillin]. AB - AIM: To analyze the related substances in ampicillin and amoxicillin and determine the related substances in products. METHODS: By using HPLC, electrospray ionzation and selective ion monitoring mass spectrum method, accelerated samples were analyzed for determination the related substances in ampicillin and amoxicillin. RESULTS: Thirteen related substances in ampicillin and 9 related substances in amoxicillin were determined. CONCLUSION: The results are useful to quality control and stability study for ampicillin and amoxicillin. PMID- 12579977 TI - [Determination of bambuterol in human plasma by liquid chromatography electrospray tandem mass spectrometry: application to pharmacokinetic study]. AB - AIM: To develop a sensitive, specific and accurate method for quantifying bambuterol in human plasma and to study pharmacokinetics of bambuterol in male healthy Chinese. METHODS: Plasma samples were prepared based on a simple liquid liquid extraction. The extracted samples were analyzed on liquid chromatography using a Zorbax SB C18 column interfaced with a triple quadrupole tandem mass spectrometer and detected by use of selected reaction monitoring mode. RESULTS: The linear calibration curves were obtained in the concentration range of 0.05 4.0 ng.mL-1. The limit of quantification was 0.05 ng.mL-1. The intra- and inter run precision was measured to be below 7%. The inter-run accuracy was less than 8% for the analyte. After an oral administration of 10 mg bambuterol hydrochloride to 18 healthy Chinese volunteers the main pharmacokinetic parameters of bambuterol were as follows: Tmax was (2.3 +/- 1.3) h; Cmax was (3.95 +/- 2.20) ng.mL-1; T1/2 was (11.4 +/- 6.1) h and AUC0-t was (26.85 +/- 11.77) ng.h.mL-1. CONCLUSION: The method is shown to be accurate, robust and convenient, and suitable for pharmacokinetic studies of bambuterol. It was found that there was marked inter-individual difference in the pharmacokinetics of bambuterol in Chinese volunteers after a single oral dose, which may be attributed to the difference of activity of cholinesterase, an enzyme catalyzing bambuterol metabolism. PMID- 12579978 TI - [Intravitreally injectable poly (D, L-Lactide) microspheres containing dexamethasone acetate for sustained release]. AB - AIM: To prepare and evaluate dexamethasone acetate-loaded poly (d, l-lactide) microspheres for sustained release in vitro and their therapeutic effect on proliferative vitreoretinopathy in vivo. METHODS: The microspheres were prepared by a solvent evaporation/extraction technique. Properties such as geometric mean diameter, span, drug loading, rate of entrapment and release characteristics were evaluated. Differential scanning calorimetry and X-ray powder diffractometry were used to identify the physical phase of dexamethasone acetate in poly (d, l lactide) matrix, and safety in vivo was examined. Effect of drug-loaded microspheres on suppression of experimental proliferative vitreoretinopathy was studied in albino rabbit eyes after an intravitreal injection of macrophages. RESULTS: The geometric mean diameter and span of the microspheres were 62.9 microns and 0.92, respectively. Drug loading and rate of entrapment of dexamethasone acetate in microspheres were 17.5% and 86.5%, respectively. Differential scanning calorimetry and X-ray powder diffractometry showed that dexamethasone acetate dispersed uniformly as molecules in poly (d, l-lactide) matrix. Ninety percent of the dexamethasone acetate was released in vitro from dexamethasone acetate after 48 days, while the same amount released from microspheres took 90 days. Storage under 4 degrees C in refrigerator or 25 degrees C in a dessicator at a relative humidity of 75% for 90 days had little effect on the properties of the microspheres. Intravitreal injection of microspheres showed a sustained release and continuous action of dexamethasone acetate for 84 days. After injection of activated macrophages, the groups of blank microspheres and dexamethasone acetate showed proliferation to different extent, while the base of the eye remained clear and no obvious proliferation was observed for the group of microspheres. A significant difference exists between these three groups. CONCLUSION: Poly (d, l-lactide) microspheres containing dexamethasone acetate is a potentially promising delivery system for the suppression of proliferative vitreoretinopathy. PMID- 12579979 TI - [Study on brain targeting 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine pharmacosomes]. AB - AIM: To investigate the specific brain drug targeting of 5-fluoro-2'-deoxyuridine (FUdR) by synthesis of 3', 5'-dioctanoyl-5-fluoro-2'-deoxyuridine (DO-FUdR) and incorporation into DO-FUdR pharmacosomes (DO-FUdR-PS). METHODS: DO-FUdR-PS was prepared by thin-layer ultrasonication technique. In vitro drug release was studied in pH 7.4 phosphate-buffered saline containing 0.3% pancreatic enzyme at 37 degrees C using bulk-equilirium reverse dialysis bag technique. The concentration of FUdR in various organs were determined by reversed phase HPLC after i.v. administration of DO-FUdR-PS and FUdR. RESULTS: The mean particle size of DO-FUdR-PS was 76 nm with drug loading of 29.02% and entrapment efficiency of 96.62%. The in vitro drug release kinetics could be characterized by bioexponential equation. Compared with FUdR injection, DO-FUdR-PS showed high concentration in tested organs. The brain AUC ratio of DO-FUdR-PS to FUdR was 10.97. CONCLUSION: DO-FUdR-PS showed a good targeting efficiency in vivo. PS can improve the ability of drug to cross blood-brain barrier and is a promising drug targeting system for the treatment of central nervous system disorders. PMID- 12579980 TI - [rDNA its sequencing of Herba Dendrobii (Huangcao)]. AB - AIM: To study the genetic diversity of ITS sequences of Herba Dendrobii (Huangcao) and analyze the utility of ITS sequences in molecular authentication of Herba Dendrobii (Huangcao) and phylogenetic of Dendrobium. METHODS: The ITS gene fragment was amplified using a pair of primers. The PCR products were purified and sequenced by the methods of Sanger Dideoxy. RESULTS: The DNA sequence of 228-233 bp ITS 1, 242-247 bp ITS 2 gene fragments and 5.8S rDNA were obtained from 14 samples of Dendrobium. The interspecific substitution varies from 11.79% to 31.58% at ITS 1 and 10.29% to 25.30% at ITS 2. The intraspecific substitution of D. nobile is 0.87% at ITS 1 and without difference at ITS 2. The substitution between Dendrobium and outgroup varies from 23.56% to 36.89% at ITS 1 and 26.52% to 33.31% at ITS 2. The phylogenetic tree based on ITS 1 and ITS 2 data was set up. CONCLUSION: The ITS 1 and ITS 2 gene fragments were highly conservative at intraspecific level in Dendrebium, while they were less conservative at interspecific level in D. nobile. They were least conservative between Dendrobium and outgroup. Hence, the sequence of this fragment is a good molecular marker for authentication of the Huangcao. But, further study is necessary for the phylogenetic of Dendrobium. PMID- 12579981 TI - [Synthesis of benzodihydropyran derivatives and evaluation of their preliminary biological activities on bone and vascular tissues]. AB - AIM: To screen optimal drugs against postmenopausal osteoporosis with cardiovascular protective activities. METHODS: A series of benzodihydropyran derivatives were designed and synthesized in view of comprehensive observations of raloxifene and ipriflavone. The antiosteoporosis activities of compounds a-e (10(-7) mol.L-1) on the proliferation of human osteoblast cell HOS TE85 were studied. The cardiovascular protective activities were evaluated by observing their effects on proliferation of human vascular endothelium cell ECV-304 and their protective effects on ECV-304 damaged by H2O2. RESULTS: Their structures were determined by spectrums. Compounds a, b and c (10(-7) mol.L-1) were shown to significantly help proliferation of HOS TE85. In addition, b, d and e (10(-8) mol.L-1) helped proliferation of ECV-304 significantly. Compounds b and c (10(-6) mol.L-1) showed strong protective activity on ECV-304 damaged by H2O2. Compounds b and c shifted the KCl dose-response curves to the right and decreased the maximal response. CONCLUSION: Compounds b and c showed some bone and vascular protective activities which benefit postmenopausal and cardiovascular diseases. PMID- 12579982 TI - [Study on the structural property of polysacchrides 139A--a kind of IL-1R antagonists from microorganism]. AB - AIM: To study the structural property of the bioactive component from the fermentation broth of Streptomyces 139 which can inhibit IL-1R. METHODS: Polysaccharide 139A is extracted and purified by means of macroreticular resin Diaion HP-20, cation exchange and DEAE Sephadex A-25 chromatography. The sugar compositions were analyzed with GC, the linkage positions of 139A were determined by methylation and GC-MS. RESULTS: The bioactive component which can inhibit IL 1R is a kind of polysaccharides. CONCLUSION: The components of 139A repeat units are conjectured. PMID- 12579983 TI - [RP-HPLC method for determination of protopine in plasma and pharmacokinetics in rats]. AB - AIM: To develop a reversed phase high performance liquid chromatographic method (RP-HPLC) for determination of protopine (Pro) in rat plasma and to investigate the pharmacokinetics of Pro in rats. METHODS: The column was packed with 5 microns C18. The mobile phase (pH 5.6) was a mixture of methanol-water-10% acetic acid (80:20:2). After twice extracted with ether under basic condition, and reextracted with 0.02 mol.L-1 sulfuric acid, protopine in the plasma samples was isolated well. The content of protopine in the plasma sample was measured by UV detector at 285 nm. RESULTS: The lowest limit of detection was 50 ng.mL-1. The intraday and interday precisions were 1.5%-3.0% and 2.1%-6.2%, respectively. The mean recovery was 80.6%-97.6%. A good linear relationship between the peak height and the concentration of protopine in rat plasma was observed. The pharmacokinetics of protopine had been investigated in rats after intravenous administration 10 mg.kg-1. The concentration-time curve of protopine in rat was confirmed to two-compartment open model. The T1/2 alpha, T1/2 beta, Ke, CL, Vd were 0.05 h, 1.85 h, 1.52 h, 6.41 L.h-1 and 17.27 L, respectively. CONCLUSION: This method is suitable for studies on pharmacokinetics of protopine. PMID- 12579984 TI - [Studies on the stability of ligustilide with solvent effect]. AB - AIM: To study the stability of ligustilide with solvent effect and try to disclose the relationship between its stability and solvent effect, so as to develop a useful method for its preservation. METHODS: The model of PCM solvent effect was used to carry out quantum chemistry calculation. The GC/MS spectrum was used to study the stability of ligustilide in solvent at room temperature. RESULTS: Ligustilide is more stable in cyclohexane and chloroform than in air. The isomerization rate of ligustilide decreased remarkably to 1.6% and 6.7% in cyclohexane and chloroform compared to 58% in air. CONCLUSION: The stability of ligustilide is greatly improved with the solvent effect. Keeping ligustilide in proper organic solvent is helpful to prevent its isomerization. PMID- 12579985 TI - [Advances in the new anti-HIV drugs acting on chemokine receptor]. PMID- 12579986 TI - Recommendations for the reporting of tissues removed as part of the surgical treatment of common malignancies of the eye and its adnexa. PMID- 12579987 TI - Recommendations for the reporting of lymphoid neoplasms. PMID- 12579988 TI - Nodular lymphocyte predominant Hodgkin lymphoma. An immunophenotypic reappraisal based on a single-institution experience. AB - In our experience, certain commonly cited immunophenotypic features of nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) are not encountered in day-to-day practice. We reviewed 60 cases of NLPHL (18 women, 42 men; median age, 34 years) to discern immunophenotypic features from a large, single-institution cohort. All cases contained lymphocytic and histiocytic (L&H) cells. These cells expressed CD20 in 98% (59/60), CD79a (usually faint) in 87% (27/31), CD30 in 7% (4/59), epithelial membrane antigen in 21% (12/56), bcl-2 in 5% (2/41), and bcl-6 in 83% (30/36) of cases. CD10 was negative in all 36 cases studied; 100% of cases (55/55) demonstrated CD3+ rosettes. Although CD57+ T cells were common within the background infiltrate, CD57+ rosettes were seen in only 48% of cases (15/31) and were rare when encountered. Based on these patterns, we conclude that bcl-2 and bcl-6 may be useful additions to the immunophenotypic analysis of NLPHL, but that the diagnostic usefulness of epithelial membrane antigen and CD57 rosettes may have been overemphasized in previous reports. PMID- 12579989 TI - Comparison of fascin expression in anaplastic large cell lymphoma and Hodgkin disease. AB - Diagnostic difficulties sometimes arise in distinguishing anaplastic large cell lymphoma (ALCL) from Hodgkin disease (HD), especially the syncytial variant. Study of the biologic features of diagnostic Reed-Sternberg cells in HD, in search of specific markers for Reed-Sternberg cells, has suggested fascin as a relatively specific and sensitive marker. We studied the frequency of fascin expression in 30 ALCLs and 34 cases of classic HD, including 17 cases of the syncytial variant. Staining with CD30 and anaplastic lymphoma kinase (ALK)-1 also was performed in all cases. All ALCL and HD cases showed membranous and Golgi zone CD30 positivity. Fascin stained all HD cases but also stained 67% (20/30) of the ALCLs in a cytoplasmic pattern. Fascin positivity was observed in 59% (10/17) of T-cell ALCLs and 77% (10/13) of null-cell ALCLs; ALK-1-positive ALCLs, regardless of origin, were usually fascin-positive (91% [10/11]). In conclusion, fascin shows strong positivity in all cases of classic HD but also is positive in the majority of ALCLs, including ALK-1-positive and ALK-1-negative cases. Positive staining for fascin is not useful for distinguishing ALCL from HD. In some cases, fascin negativity may help rule out classic HD. PMID- 12579990 TI - Immunophenotypic analysis of anaplastic large cell lymphoma by flow cytometry. AB - We studied the antigen expression profiles of 19 anaplastic large cell lymphoma (ALCL) cases by multiparameter flow cytometry. The neoplastic cells expressed CD45, HLA-DR, and CD30 in all cases. At least 1 T cell-associated antigen was expressed in each case (CD2, 12/17 [71%]; CD4, 12/19 [63%]; CD3, 6/19 [32%]; CD7, 6/19 [32%]; CD5, 5/19 [26%]; CD8, 4/19 [21%]). CD25 was expressed in 14 (88%) of 16 cases. CD13 was expressed unexpectedly in 8 (47%) of 17 cases. One CD13+ ALCL also was positive for CD33 and 2 others for CD15, CD19, CD20, CD22, CD14, and CD36 were not expressed. Anaplastic lymphoma kinase protein was detected in about 33% (3/9) of ALCLs examined by flow cytometric immunophenotyping (FCI); expression was validated by immunohistochemical analysis. Of 19 ALCL cases, 12 were diagnosed solely based on FCI findings in conjunction with morphologic evaluation of body fluid (1 case), fine-needle aspirate (3 cases), or excisional biopsy specimen (8 cases). The diagnoses of the remaining 7 cases were suggested strongly by FCI and confirmed by immunohistochemical analysis. FCI is useful to aid in diagnosis of ALCL, particularly along with fine-needle aspiration evaluation. ALCL with aberrant expression of myeloid antigens should not be mistaken for extramedullary myeloid tumor. PMID- 12579991 TI - Minimal residual disease detection in hairy cell leukemia. Comparison of flow cytometric immunophenotyping with clonal analysis using consensus primer polymerase chain reaction for the heavy chain gene. AB - We studied 86 specimens from 24 patients with hairy cell leukemia (HCL) to determine the sensitivity of routine flow cytometry (FC) and consensus primer PCR (cpPCR) in this disease. FC was more sensitive, detecting HCL in 48 (56%) of 86 specimens, while clonal B-cell populations were detected by cpPCR in only 23 (27%) of 86 specimens. FC and cpPCR were both more sensitive than morphologic examination. A positive cpPCR result is associated with higher tumor cell numbers than a negative cpPCR result, as determined by FC (P = .0017). We determined cutoff values for number of tumor cells at which cpPCR is consistently positive. At 6.8 tumor cells per microliter, cpPCR would be expected to be positive in at least 90% of the samples. FC was adequate in 86 cases (100%), while cpPCR was adequate in 74 cases (86%). FC is superior to cpPCR for detecting minimal residual HCL. It is more sensitive and more specific and permits quantitation of tumor cell number. PMID- 12579992 TI - B-cell lymphomas with coexpression of CD5 and CD10. AB - Coexpression of CD5 and CD10 is highly unusual in B-cell lymphomas and may pose a diagnostic challenge. We report 42 cases of B-cell lymphoma with simultaneous expression of CD5 and CD10. They made up approximately 0.4% of all B-cell lymphomas seen during the study period and included the following cases: large B cell lymphoma (LBCL), 14 (33%); follicular lymphoma (FL), 10 (24%); mantle cell lymphoma (MCL), 9 (21%); chronic lymphocytic leukemia, 4 (10%); acute precursor B cell lymphoblastic leukemia/lymphoma, 2 (5%); and other low-grade B-cell lymphomas, 3 (7%). All MCLs had overexpression of bcl-1 or the t(11;14) and were CD43+. All FLs had typical histomorphologic features and were bcl-2+ and bcl-6+ but CD43-. Of 14 LBCLs, 5 were histologically high-grade. Six (43%) of 14 patients with LBCL died within 10 months of diagnosis of CD5+CD10+ lymphoma (median survival, 4 months), including all 3 patients with stage IV disease and 2 of 5 with histologically high-grade lymphoma. Our findings indicate that coexpression of CD5 and CD10 is rare but occurs in diverse subtypes of B-cell lymphoma. Investigation of bcl-1, bcl-6, and CD43 and morphologic evaluation may resolve the potential confusion in diagnosis and lead to the recognition of the correct lymphoma subtype. PMID- 12579993 TI - European concerted action on anticoagulation. Minimum numbers of lyophilized plasma samples for ISI calibration of CoaguChek and TAS point-of-care whole blood prothrombin time monitors. AB - International sensitivity index (ISI) calibration of whole blood prothrombin time (PT) monitors is too complex. We previously simplified the method by using European Concerted Action on Anticoagulation (ECAA) lyophilized plasma samples with the TAS PT-NC (Bayer AG, Leverkusen, Germany) and the CoaguChek Mini (Roche Diagnostics, Mannheim, Germany) whole blood PT monitoring systems. The TAS PT-NC required a correction derived from the line of equivalence. Monte Carlo bootstrap analysis of reducing numbers of test samples was performed with both systems. Plasma samples from patients receiving coumarin (coumarin samples), healthy subjects (normal samples), and plasma samples artificially depleted of coagulation factors were used. With the TAS PT-NC, 20 coumarin samples or 20 artificially depleted samples with 7 normal samples gave reliable ISI and international normalized ratio and satisfactory precision. With the CoaguChek Mini, 30 coumarin and 10 normal samples were required. Simplification of ISI calibration of the 2 monitoring systems is possible using fewer ECAA lyophilized plasma samples than the 80 required according to the World Health Organization guidelines for conventional PT systems and previously recommended for fresh plasma samples tested on the same 2 monitoring systems. PMID- 12579994 TI - International sensitivity index calibration of the near-patient testing prothrombin time monitor, ProTime. AB - Near-patient testing devices (monitors) capable of measuring prothrombin time on an unmeasured drop of blood would be suitable alternatives to centralized laboratory monitoring of patients receiving oral anticoagulants. The essential prerequisite for the use of these monitors is their conformity to the international sensitivity index (ISI) calibration model recommended by the World Health Organization. We report on the ISI calibration of the ProTime monitor (International Technidyne, Edison, NJ) designed and approved for patient self testing. According to our results, this monitor can be calibrated by adopting the model already used for other monitors. The apparent ISI was close to unity. Overall, the international normalized ratio values displayed by the monitor agreed with those measured with the international reference preparation for thromboplastin. Confirmation of these results in a large multicenter study is warranted. PMID- 12579995 TI - Atypical ductal hyperplasia diagnosis by directional vacuum-assisted stereotactic biopsy of breast microcalcifications. Considerations for surgical excision. AB - In 824 patients who underwent directional vacuum-assisted biopsies (DVABs) of breast microcalcifications, 61 (7.4%) showed atypical ductal hyperplasia (ADH). The 42 who subsequently underwent excision were the subjects of this study. Cases were evaluated for the mammographic characteristics of the lesion, the percentage of lesion removed according to mammography, and histologic findings (including number of large ducts and/or terminal duct-lobular units involved with ADH) in DVAB specimens. Pathologic findings in the surgical specimens in the area of the biopsy site also were recorded. In the DVAB specimens, ADH was confined to an average of 1.5 large ducts or lobular units and was associated with microcalcifications in all of the patients. Surgical specimens showed ADH in 15 cases, no residual lesion in 24 cases, and ductal carcinoma in situ in 3 cases. We found that microcalcifications that contain ADH in less than 3 lobules or ducts and/or that are removed completely by DVAB do not reveal higher-risk lesions on excision; thus, removal is unnecessary. When assessing microcalcifications with ADH, clinicians should consider the percentage of microcalcifications removed by DVAB and the extent of lobular involvement to better assess the need for excision. PMID- 12579996 TI - Stellate-cell lipidosis in liver biopsy specimens. Recognition and significance. AB - Hepatic stellate-cell lipidosis due to hypervitaminosis A can lead to cirrhosis, which can be averted by restricting vitamin A intake. Other causes, including the use of synthetic retinoids, have been postulated. We studied the frequency and etiology of stellate-cell lipidosis in patients undergoing liver biopsy for reasons other than vitamin A abuse. Fourteen cases (1.1%) were identified retrospectively among 1,235 nontransplant liver biopsy specimens examined from January 1995 through December 1999. Diagnostic criteria included the following: lipid-laden cells in the space of Disse; small, dark, crescent-shaped nuclei with inconspicuous nucleoli; and wispy cytoplasmic strands separating fat droplets. Patient details, reason for biopsy, and medication use were studied. Reasons for biopsy included hepatitis C (10 cases), abnormal liver enzyme levels (2 cases), methotrexate use (1 case), and alcohol abuse (1 case). Hypervitaminosis A was not suspected clinically in the 5 patients who used oral vitamin A or 3 who used topical tretinoin (Retin-A). In 6 patients, no cause of stellate-cell lipidosis was discerned. Stellate-cell lipidosis should be reported to alert clinicians to a potentially preventable form of liver injury. PMID- 12579997 TI - Expression of calretinin and the alpha-subunit of inhibin in granular cell tumors. AB - Granular cell tumors (GCTs) typically express S-100 protein, which has been used as a marker in differential diagnosis. Calretinin, a calcium-binding protein related structurally to S-100, and inhibin, a polypeptide hormone secreted primarily by ovarian granulosa cells and testicular Sertoli cells and functioning as an inhibitor for pituitary follicle-stimulating hormone secretion, are potentially useful but not well-evaluated markers for GCTs. We studied 43 cases of GCT with antibodies to calretinin, the inhibin alpha-subunit, and S-100 protein. All tumors were positive for inhibin alpha-subunit and S-100 protein, with 50% or more cells showing moderate to strong staining. Forty tumors (93%) were positive for calretinin, ranging from focal weak to diffuse strong staining. Enhanced staining for calretinin in the tumor cells adjacent to hyperplastic squamous epithelium was observed in 9 of 13 cases showing pseudoepitheliomatous hyperplasia. Calretinin and the inhibin alpha-subunit are useful markers for GCTs. The expression of calretinin, a primarily neuronal protein, in GCTs further supports its neural differentiation or derivation. The elevated calretinin expression in the tumor cells adjacent to the hyperplastic squamous epithelium suggests a role for calretinin in the tumor cells-squamous epithelium interaction. PMID- 12579998 TI - Comparative genomic hybridization. Synchronous occurrence of focal nodular hyperplasia and hepatocellular carcinoma in the same liver is not based on common chromosomal aberrations. AB - Occasionally hepatocellular carcinomas (HCCs) occur synchronously with or within focal nodular hyperplasias (FNHs), raising the question of a putative causal relationship. In the present study, we used comparative genomic hybridization to investigate the occurrence of genomic aberrations in FNHs, which might lead to hepatocarcinogenesis. Tissue samples from FNHs and nonlesional liver tissue were obtained from 7 women. None of the patients had a chronic diffuse liver disease. A synchronous HCC not spatially related to FNH was present in 1 patient. Two patients had received oral contraceptives. Genomic aberrations were found in only 1 FNH. No aberration was found in the FNH occurring synchronously with HCC, but the HCC included gains at chromosomes 1q, 5, 12, and 19q and losses at 4p, 7q22 q35, 9p, 17p, 21q, and 22q. No aberrations were found in nonneoplastic liver tissues. Our findings support the notion that FNH is not a preneoplastic lesion for the occurrence of HCC in humans and that the synchronous occurrence of FNH and HCC is coincidental in our case. PMID- 12579999 TI - Quantitative analysis of serum free light chains. A new marker for the diagnostic evaluation of primary systemic amyloidosis. AB - Primary systemic amyloidosis is a plasma cell dyscrasia characterized by the accumulation of excess free immunoglobulin light chains (FLCs) as amyloid. One of the diagnostic features of amyloidosis is the presence of circulating monoclonal FLCs in the serum and urine of the patients. The FLC usually is present in small amounts, and immunofixation is required for detection. A nephelometric method for quantitating FLCs in serum has been described using antibodies that recognize only FLC not bound to heavy chain. We describe a retrospective study using this quantitative FLC method for assessing monoclonal FLCs in 95 patients with amyloidosis. The sensitivity of nephelometric serum FLC measurements is particularly useful in patients with negative immunofixation results for serum, urine, or both. In addition, the FLC assay can be used for follow-up of patients with amyloidosis who have undergone stem cell transplantation. PMID- 12580000 TI - Direct and indirect immunofluorescence as a diagnostic adjunct in the interpretation of nonneoplastic medical lung disease. AB - Fresh open lung biopsy material from 57 patients was incubated with fluoresceinated complement and immunoglobulin antisera. An indirect immunofluorescent assay using neonatal lung as substrate was conducted as well. Direct immunofluorescent patterns could be categorized into interalveolar septal capillary deposition, large vessel wall localization, alveolar basement membrane localization, or a pauci-immune immunofluorescence pattern. With respect to the septal capillary pattern, endothelial cell decoration was seen with scleroderma, mixed connective tissue disease, anti-Ro-associated lupus erythematosus, dermatomyositis, humoral allograft rejection, and patients with isolated pulmonary fibrosis in whom autoantibodies were established, including antiphospholipid antibodies. A similar pattern of endothelial cell staining was seen in these cases via the indirect assay. Granular mural septal capillary deposition was seen in the aforesaid settings along with rheumatoid factor positive rheumatoid arthritis, type II cryofibrinogenemia, and mixed cryoglobulinemia and, in some cases, light microscopically corresponded to a neutrophilic capillaritis. Isolated vascular IgA corresponded with rheumatoid arthritis corresponding to IgA-specific antiendothelial cell antibodies, celiac disease-associated pulmonary hemorrhage, Schonlein-Henoch purpura and with IgA antiphospholipid antibodies. Alveolar wall deposition was seen with anti glomerular basement membrane disease. PMID- 12580001 TI - Effects of sterilizing gamma irradiation on bloodspot newborn screening tests and whole blood cyclosporine and tacrolimus measurements. AB - Sterilizing irradiation of the US mail has been proposed as a method to prevent delivery of viable anthrax spores. Because newborn screening samples (bloodspots) and cyclosporine and tacrolimus specimens (whole blood) are delivered routinely through the mail, we studied whether sterilizing gamma irradiation could affect these test results. Specimens were exposed to 18 kGy gamma irradiation (100 hours x 18,000 rad/h), a "kill dose" for Bacillus pumilus spore strips. Irradiation had no significant effect on whole blood cyclosporine or tacrolimus results, but it had a degradative effect on bloodspot phenylalanine, hemoglobins, biotinidase, galactose-1-phosphate uridyltransferase, thyroxine, and thyrotropin. Such irradiation potentially could cause false-negative results for the detection of phenylketonuria and likely would lead to an increase in secondary testing for hemoglobin variants, but it is unlikely to lead to false-negative or false positive results for the remaining newborn screening tests. These experiments cannot rule out possible greater effects by larger doses or other types of irradiation. PMID- 12580003 TI - [The psychiatric care is charged with value conflicts. Unclear dividing-line between the voluntary and mandatory care]. PMID- 12580002 TI - Effect of Asian and Siberian ginseng on serum digoxin measurement by five digoxin immunoassays. Significant variation in digoxin-like immunoreactivity among commercial ginsengs. AB - Asian and Siberian ginsengs contain glycosides with structural similarities to digoxin. We studied potential interference of ginseng in 5 digoxin immunoassays in 3 Asian (2 liquid extracts, 1 capsule) and 3 Siberian ginseng preparations (1 liquid extract, 2 capsules). With the fluorescence polarization immunoassay (FPIA), we observed apparent digoxin activity in 1 Asian liquid preparation and in the liquid extract and 1 capsule form of Siberian ginseng. In mice fed ginseng, we observed digoxin activities in the serum (Asian, 0.48-0.68 ng/mL [0.6 0.9 nmol/L]; Siberian, 0.20-0.47 ng/mL [0.3-0.6 nmol/L]), indicating that such interferences also occur in vivo. Serum pools prepared from samples from patients receiving digoxin and then supplemented with Asian or Siberian ginseng showed falsely increased digoxin values using the FPIA (e.g., for Asian ginseng, 1.54 ng/mL [2.0 nmol/L] vs control value, 1.10 ng/mL [1.4 nmol/L]) and falsely decreased values using the microparticle enzyme immunoassay (MEIA; 0.73 ng/mL [0.9 nmol/L] vs control value, 1.04 ng/mL [1.3 nmol/L]). Digoxin-like immunoreactive substances (DLISs) showed synergistic effects with ginsengs in interfering with the FPIA and MEIA for digoxin. No interference was observed with 3 other digoxin assays, even in the presence of elevated DLISs. PMID- 12580004 TI - [Experiences from a new surgical unit. New approach in prolapse surgery has eliminated the waiting list]. AB - A unit specializing in the surgical repair of vaginal vault prolapse was formed. The objective was to treat all kinds of defects in local anaesthesia with a hospital stay not exceeding 24 hours. Almost 800 women have been operated upon in the manner outlined in less than 26 months time. All but nine patients were dismissed as scheduled. Morbidity was low. PMID- 12580005 TI - [Shortages of population studies according a survey. Questions concerning skin allergy and hypersensitivity should be coordinated and unified]. PMID- 12580006 TI - [Depleted uranium a cancer risk that disappeared. Leukemia alarm regarding Balkan veterans came to nothing]. AB - After alarming reports in the international press in January 2001, about leukemia in war veterans returning from the Balkans after possible exposure to depleted uranium, a follow-up was conducted of the Swedish personnel that had served in the Balkans. Questionnaires, analysis of uranium in urine, and coordination with The National Board of Health and Welfare's cancer register showed no correlation between service in the Balkans and cancer or other illnesses. Several did however experience anxiety, insomnia and fatigue that may have been caused by the stressful environment and/or the anxiety arising from the depleted uranium debate. To lower the risk for unjustified anxiety and to be better prepared for the physical environment, the Swedish Armed Forces are working on better risk analysis before mission as well as increased health examinations both before and after mission. PMID- 12580007 TI - [Parvovirus B19 infection--a risk factor during pregnancy. Asymptomatic course of disease makes the prevention and treatment more difficult]. PMID- 12580008 TI - ["Contract" prevents school youth from initiating tobacco use. Percentage of smoking/snuff-taking 13-16-year-olds cut in half over a six-year period]. AB - A contract has been used as an aid to prevent tobacco use among pupils in certain schools in Sweden. A local variant of this contract ("Antidrogkortet") ("Drug resistance card") has been used in the community of Habo in 6th to 9th classes in the nine-year compulsory school since 1995. By signing the contract the pupil promises not to use tobacco during the present school year. The contract also has a function of a lottery ticket where the winnings are distributed twice each term. An evaluation has been made by means of a questionnaire to the pupils. A third of the pupils who answered the questionnaire stated that the contract has helped them not to use tobacco. Since 1995 the number of smokers in 7th to 9th classes has decreased from 16% to 7%, and the number of users of moist snuff has decreased from 8% to 4%. PMID- 12580009 TI - [Too many methods are used for measurement of health-related quality of life]. PMID- 12580010 TI - [Is there a right of being crazy? The most severely mentally ill persons are failed by the society and a compliant compulsory care legislation]. PMID- 12580011 TI - [Henri Michaux wrote and painted "to keep his demons in check"]. PMID- 12580012 TI - [The responsibility is put on physicians in spite of a glamorous rhetoric around priorities and care guarantees]. PMID- 12580013 TI - [A reply to Per Gunnar Holmgren: Is the placement of advertising in the Dagens Medicin random or controlled?]. PMID- 12580014 TI - [A clever idiot?]. PMID- 12580015 TI - [Shoot the messenger!--or the art of changing the subject of conversation]. PMID- 12580016 TI - [Why this arrogance, Josef Milerad?]. PMID- 12580017 TI - [The study grant system is not connected to reality]. PMID- 12580018 TI - Peripheral nerve and transgene cells transplantation in the treatment of experimental neuropathy of SD rats. AB - PURPOSE: To observe whether optic nerve can regenerate by peripheral nerve transplantation and GDNF gene transfer. METHODS: Optic nerve was transected about 2 mm posterior to the eye to establish experimental models of optic neuropathy. Sciatic nerve was sutured to the sheath of injury optic nerve by a 10-0 nylon, pcDNA3-GDNF was smeared to the joint and injected to the vitreous cavity. The regenerated retinal ganglion cells axons were observed by HRP. The retina and optic nerve were observed by pathohistological methods. RESULTS: Some retinal ganglion cells with regenerated axons could be seen a week after transplantation with large cell bodies. The cells could be identified by HRP. The regenerated nerve fibers grew to the transplanted sciatic nerve. Then the number of retinal ganglion cells with regenerated axons increased with time going. Only a few regenerated retinal ganglion cells could be seen two months after transplantation. The large retinal ganglion cells were seen to have plenty of cytoplasms and be in an active state under electron microscope. Some nerve fibers could be seen in the joint. Sciatic nerve transplantation combined with GDNF gene therapy could increase the amount of retinal ganglion cells with regenerated axons compared with those without GDNF transgene. CONCLUSIONS: Optic nerve could regenerate by peripheral nerve transplantation and GDNF gene transfer. PMID- 12580019 TI - The saturation characteristics of glucose transport in bovine retinal pigment epithelium. AB - PURPOSE: To study the saturation characteristics of the glucose transport across the bovine retinal pigment epithelium(RPE). METHODS: The bovine RPE preparations were munted with a modified Ussing chamber. The L-[3H]-glucose and 3-O-methyl-D [14C]-glucose fluxes across the RPE from the choroid to retina were studied at different glucose concentrations. RESULTS: The glucose transport was found to be stereospecific, with 3-O-methyl-D-glucose (MDG) being transported about three times faster than L-glucose. The glucose transport showed typical saturation characteristics in Michaelis-Menten fashion. The Vmax and the Km of corrected MDG were 2452 nmol cm-2h-1 and 30.8 mM respectively. It was shown that the glucose transport system was saturated at 61.6 mM. CONCLUSIONS: The saturation characteristics of the corrected MDG flux suggested that the capacity of glucose transport through the bovine RPE is immense. PMID- 12580020 TI - Color vision defects with variation in the exon 5 of red and green pigment genes. AB - PURPOSE: To investigate correlation of variation in the exon 5 of red and green pigment genes with color vision defects. METHODS: Exon 5 of the red and green pigment genes in 11 protans, 19 deutans and 38 normal controls were analyzed by heteroduplux-SSCP analysis. RESULTS: In all 11 protans and 8 of the 19 deutans, defects of the red or green pigment gene could be identified. The C polymorphism (A/C at codon 283) in green pigment gene was present in 8 of 44 trichromats and 5 of 24 dichromats. Specific electrophoretic bands were found in 2 normal controls and a deutan. CONCLUSIONS: Variation in the exon 5 of the red and green pigment genes is the most common cause for color vision defects. Heteroduplex-SSCP analysis is a suitable way in screening specific variation in visual pigment genes. PMID- 12580021 TI - The study on culture and ultrastructure of glaucomatous trabecular cells in vitro. AB - PURPOSE: To establish the culture system of human glaucomatous trabecular cells in vitro and study their ultrastructures. METHODS: The trabecular specimens from trabeculectomy were cultured in vitro and passaged 3 times, then identified. Moreover, the glaucomatous cells were observed with electron microscope while compared with the normal ones. RESULTS: Cultured human glaucomatous trabecular cells were obtained. The ultrastructure of the cells showed the decrease in vilious project, coated vesicle and lysosomal inclusion. CONCLUSION: The establishment of human glaucomatous trabecular cells culture in vitro made the culture system more perfect. The morphologic changes might be related to the abnormal functions of human trabecular meshwork cells. PMID- 12580022 TI - Hemodynamics of ophthalmic artery and central retinal artery and correlation with other factors in patients with primary open angle glaucoma. AB - OBJECTIVE: To investigate the hemodynamics of ophthalmic artery (OA) and central retinal artery (CRA) in patients with primary open angle glaucoma(POAG) and analyse the correlation between hemodynamics of OA and CRA and other factors. METHODS: The hemodynamics of OA and CRA in normal persons (102 eyes) and glaucomatous patients (102 eyes) were measured by Colour Doppler Image (CDI). The linear correlation analysis was made between OA and CRA about each of the following parameters: the peak systolic flow velocity (Vmax), the end diastolic velocity (Vmin). The multiple stepwise regression analysis was taken to investigate the correlation between each of the following hemodynamics of OA and CRA: Vmax and Vmin in 74 eyes with POAG and each of the following related factors in hemorrheology: whole blood apparent viscosity at low, medium and high shear rate, plasma viscosity and hematocrit. The multiple linear regression was applied to investigate the correlation between each of the following hemodynamics of OA and CRA: Vmax and Vmin in 69 eyes with POAG and each of the following blood vessel filling time: the arm-retinal artery (A-AT) and retinal artery-venous (A VT) of the fundus fluorescein angiography(FFA). RESULTS: The Vmax, Vmin and time averaged maximum velocity (Vmean) of OA and CRA in patients with POAG were lower than those in normal persons, but the resistance index (RI) of OA and CRA was higher than those in normal. There was a positive correlation between OA and CRA in Vmin, but a negative correlation between Vmin of OA and plasma viscosity. The whole blood apparent viscosity at low shear rate was clodely related to Vmax and Vmin of CRA. Also there was a negative correlation between Vmax of CRA and A-AT of the FFA. CONCLUSION: There was abnormal hemodynamics, which is influenced by blood viscosity, of OA and CRA in patients with POAG. Abnormal hemodynamics can affect the blood supply to optic disc and retina in POAG. PMID- 12580023 TI - Characterization of glucocorticoid receptor on lymphocytes in Chinese patients with glucocorticoid-induced glaucoma. AB - PURPOSE: We studied the pathogenesis of glucocorticoid-induced glucoma (GIG) through characterization of glucocorticoid receptor (GR) on lymphocytes in Chinese patients with GIG. METHODS: By radioligand receptor binding followed by Scatchard analysis, the specific binding sites were characterized and quantitated for glucocorticoid receptors on peripheral blood lymphocytes obtained from patients with GIG and the control group. RESULTS: The binding sites we detected were as follows: 12.7 +/- 1.47 x 10(3) receptors per cell with a KD of 3.02 +/- 0.62 nmol/L in patients with GIG, 7.26 +/- 0.45 x 10(3) receptors per cell with a KD of 3.03 +/- 0.56 nmol/L in the control group. The statistical difference of receptors per cell is significant between two groups (p < 0.05), patients with GIG having more GR binding sites, while the difference of Kd is not significant (p > 0.05). CONCLUSION: The preliminary findings suggest that patients with GIG are more sensitive to glucocorticoid and the increase of binding sites of GR may be the receptor and molecular basis of the pathogenesis of GIG. PMID- 12580024 TI - Prejunctional 5-HT receptors enhance cholinergic neurosecretion in the rabbit and human iris-ciliary body. AB - PURPOSE: To characterize prejunctional 5-HT heteroreceptors which modulate 3Hacetylcholine (3H-ACh release) in isolated rabbit and human iris-ciliary bodies (ICB3). METHODS: ICB tissue segments were incubated with 3H-choline, superfused and electrically stimulated four times (S1, S2, S3, S4) at 3-10 Hz for 1 min to elicit 3H-ACh. secretion. Test agents (5-HT agonists and antagonists) were added before S2, S3 and S4 and their effects determined by the stimulation ratio (Sx/S1) of evoked 3H-ACh. release. 3H-ACh in superfusate fractions was fractionated and quantified by ion exchange chromatography. RESULTS: In rabbit ICBs, evoked 3H-ACh. release was enhanced in a concentration-dependent manner by 5-HT (10(-9)-10(-5) M, EC50 = 5.8 x 10(-8) M). The maximum effect of 5-HT (10(-6) M) corresponded to a 45.14 +/- 7.40%) (n = 6) increase in 3H-ACh release. Higher concentrations of 5-HT (> 10(-4) M) induced desensitization. The response to 5-HT (10(-6) M) in the presence of the 5-HT3/4 antagonist tropisetron (10(-9) M), 5 HT1 antagonist methiothepin ((10(-7) M), equated to 89.96% and 88.78% respectively of control value; in the presence of non-selective 5-HT antagonist mianserin, 5-HT4 antagonist SDZ-205557 (10(-7) M) was -15.20% and 32.84% of control. The 5-HT response was mimicked by the selective 5-HT4 agonist 5 methoxytryptamine (10(-9)-10(-4) M, EC50 = 7 x 10(-8) M), whereas the 5-HT3 agonist M-CPBG[(1-m-chlorophenyl)-biguanide] and phenylbiguanide, the 5-HT1 agonist 5-carboxamidotryptamine and the 5-HT2A agonist alpha-methyl-5-HT were ineffective. The selective 5-HT1A agonist (+)-8-OH-DPAT (10(-8)-10(-5) M) had no significant effect, but at concentration of 10(-4) M, inhibited evoked 3H-Ach. release. Similar results were obtained in the human ICB. The evoked 3H-Ach. release was enhanced in a concentration-dependent manner by 5-HT (10(-9)-10(-5) M, EC50 = 3.36 x 10(-8) M) or 5-MOT (10(-8)-10(-5) M, EC50 = 6.59 x 10(-7) M), The selective 5-HT4 antagonist, GR113808A (10(-8) M) inhibited the 5-HT-inducing increase of the cholinergic response, producing parallel right shits of the concentration-response curves to 5-HT. The selective 5-HT3 antagonist ondersetrone (5 x 10(-7) M) and tropisetron (10(-9) M) did not affect 5-HT inducing 3H-Ach release. CONCLUSIONS: Our results indicate that cholinergic terminals in the rabbit and human ICB contain facilitatory 5-HT heteroreceptors that belong to the 5-HT4 subtype. Their role in modulation of intraocular pressure remains to be elucidated. PMID- 12580025 TI - Mycobacterium chelonei keratitis: report of a case and review of the literature. AB - BACKGROUND: Mycobacterium (M) chelonei keratitis is a rare opportunistic eye infection that can cause significant morbidity when not being treated properly. The first case was documented by Gangadharam et al in 1978 and since then, a total of 49 cases were reported in the literature. One alarming fact is that more than 50% of cases were found in the Chinese population and mostly reported in recent years. The key to successful management of M. chelonei keratitis is early diagnosis by high index of suspicion. In order to alert ophthalmologists of this condition, we report a typical case of M. chelonei keratitis and review the literature of all the reported cases with special reference to its risk factors, treatments and outcome. METHODS: The cases reported in the literature and a case of our own were reviewed and analyzed. RESULTS: Our case was a 42-year-old gentleman who developed M. chelonei keratitis following pterygium surgery. He had typical clinical features of irregular infiltrates with radiating projections, indistinct fluffy lesion margins, satellite lesions and associated epithelial defect. Penetrating keratoplasty was performed after failed medical treatments. He recovered fully with a best corrected visual acuity (BCVA) of 20/30 at 24 months after the corneal transplant. A total of 49 cases were reported in the literature. The major risk factor was corneal injury, including surgical trauma. Corneal foreign bodies (24 cases, 48%) were found to be highly correlated, especially metallic foreign bodies (16 cases, 32%). Diagnosis was usually delayed for weeks or months and medical treatment alone often failed. Amikacin is usually the treatment of choice but its efficacy is just sub-optimal. Multi-resistance to the commonly used board spectrum antibiotics is not uncommon. The drug sensitivity test against atypical mycobacterium is technically difficult to perform and in vitro results are well known to be poorly related to clinical response. Combined extirpative keratectomy and topical antibiotics had been tried and was shown to be effective. Penetrating keratoplasty appeared to be a good definitive treatment for drug-resistance and advanced cases. CONCLUSION: M. chelonei keratitis is a rare opportunistic infection. The major risk factor is eye injury, with foreign bodies or surgical trauma. Diagnosis is often missed and delayed as a result of its scarcity and variable presentations. High index of clinical suspicion with early diagnosis and prompt combined medical and surgical intervention seem to be the best measure to decrease ocular morbidity. Good alertness and knowledge of this condition would help our patients in the Far East as the literature review has shown a recent trend of increase in frequency and more than 50% of the reported cases come from the Chinese patients. PMID- 12580026 TI - A study of clinical characteristics and treatment of Mooren's ulcer. AB - OBJECTIVE: To investigate the clinical characteristics and to compare the effects of several therapies of Mooren's corneal ulcer. METHODS: 550 consecutive cases of Mooren's corneal ulcer inpatients were analysed, including the age, sex, laterality of the eye, ulcer location, perforative rate, cure rate of surgeries, recurrent rate, and the effects of conjunctiva excision, lamellar keratoplasty (LKP), LKP and plus cyclosporin A eye drop. RESULTS: The average age of onset of the corneal ulcer was 48.4 years old, the ratio of the male to the female patients was 1:0.74, the bilateral disease was 30% of the total cases, 31.5% of the bilateral disease occurred in the younger group, and 68.5% of the bilateral ulcer occurred in the older group, ulcers located at the limbus of the palpebral fissure were 70% of the total cases, perforative rate was 13.3%, 43.2% of the perforation occurred in the younger group, and 56.8% of the perforation occurred in the older group, recurrent rate of the post-operation was 25.6%, the cure rate of the first procedure of LKP and plus cyclosporin A eye drop was 73.7%, the final cure rate was 95.6%, and the preservative rate of the corneal anatomical structure of the surgeries was 99.7%. CONCLUSION: The study primarily provided the clinical characteristics of Mooren's corneal ulcer patients of China. The available data do not support the present concept of the classification of the two clinical types. The bilateral ulcer may be the malignant type of Mooren's ulcer. LKP and plus cyclosporin A eye drop was an effective treatment of Mooren's corneal ulcer. PMID- 12580028 TI - Analysis of oscillatory potentials of flash electroretinogram in frequency domain and time domain in retinal vein occlusion. AB - PURPOSE: To analyse the parameters of frequency domain and time domain of the OPs in branch retinal vein occlusion (BRVO) and central retinal vein occlusion (CRVO). METHODS: OPs from 9 eyes of BRVO, 9 eyes of CRVO and 40 normal control eyes were tested. The frequency spectra of the OPs were derived through Fast Fourier Transform (FFT). The mean frequency spectra, the total power, the maximum magnitude, the maximum power and the dominant frequency in frequency domain of the OPs from the eyes of BRVO, CRVO and the normal control group were compared with each other. The OPs waveforms were reconstructed with the OPs frequency spectra data by counter-FFT. In time domain, the latencies, the amplitudes and the sum of the amplitudes of the four wavelets of OPs were measured and compared in the above subjects. RESULTS: There were no statistically significant differences in the magnitudes of frequency spectra, the total power, the maximum magnitude and the maximum power in the frequency range from 110 to 250 Hz between BRVO and normal subjects. However, in BRVO, the dominant frequency decreased significantly, the latencies of the first to the third OPs wavelet prolonged significantly, the amplitudes of the first and the second OPs wavelet and the summed amplitude of the four OPs wavelets attenuated significantly. Comparing the results of CRVO with those of normal subjects, there were statistically significant decrease in the magnitudes of frequency spectra from 150 to 180 Hz, there were statistically significant decrease in the total power, the maximum magnitude, the maximum power and the dominant frequency in the frequency range from 110 to 250 Hz, the latencies of the first to the fourth OPs wavelet prolonged significantly, the amplitudes of the first to the third OPs wavelet and the summed amplitude of the four OPs wavelets attenuated significantly. CONCLUSION: Some parameters of OPs in frequency domain and time domain are abnormal in branch retinal vein occlusion and in central retinal vein occlusion, which is more marked in the latter. PMID- 12580027 TI - Studies on human fetal lens crystallins under oxidative stress and protective effects of tea polyphenols. AB - PURPOSE: To investigate the oxidative modification of water-soluble crystallins of human fetal lens with H2O2 and fourteen metal ions with or without EDTA. Tea polyphenols (TP) was added to above solutions in order to testing their antioxidative abilities. METHODS: The experiments were performed at 37 degrees C with final concentration of 2.5 mg/ml protein, 0.1 mM metal ions, 0.3 mM EDTA and 1.0 mM H2O2. Then the TP was added to the solution with CuSO4 and H2O2, after 5 or 24 hours, the crystallins were analysed with SDS-PAGE and IEF. RESULTS: There were marked oxidative modifications of lens protein in H2O2 and copper without EDTA. In SDS-PAGE patterns, we found an increase in those species above of bands higher than 30 kD and some diffuse bands from 30 to 17 kD after 5 hours. In IEF patterns, there were a general increase on acidity with loss of the more basic species. When the TP was added, there was not any difference with control group. CONCLUSION: The results indicate that exposure of water-soluble protein to H2O2 and copper leads to covalent crosslinking and cleavage of polypeptides. After 24 hours, the development of the oxidative modifications of crystallins continues, comparison with the catalytic strength, copper ions stronger than the iron ions. On other hand, this work reported that the anti-oxidative action of TP is strong. PMID- 12580029 TI - Unilateral acute idiopathic maculopathy. AB - PURPOSE: To discuss the fluorescein angiographic features and diagnosis as well as differential diagnosis. METHODS: In our hospital, we have found three cases of UAIM, one of them has eccentric maculopathy, another has papillitis, and the third associates with pregnancy. Each has characteristic clinical findings. We have made the diagnosis base on ophthalmoscopy and fluorescein angiography (FA). RESULTS: Three patients with UAIM have been experienced sudden unilateral decreased visual acuity. The significant changes on fundus are retinal pigment epithelium (RPE) thickening with neurosensory retinal detachment with in the macular area. FA showed early patchy hypofluorescence and hyperfluorescence at the level of the RPE. 'Bull's eye appearance may occur in the macular area after the acute stage of the disease. The clinical findings are similar to what the foreign report described. But there were no previous reports in China. CONCLUSION: We can establish the diagnosis on the base of clinical and fundus features associated with fluorescein angiographic findings. However, the etiology and pathology could not be identified. It required further research. PMID- 12580030 TI - [The preliminary experiment of gene transfer into cultured rat retinal neural cells]. AB - PURPOSE: To investigate the possibility of gene transfer into retina neural cells. METHODS: pcDNA3-LacZ and pcDNA3-GDNF were transferred into rat retinal neural cells with cationic lipofectin. The gene transfer and expression of GDNF were detected by means of RT-PCR and SDS-PAGE. RESULTS: pcDNA3-LacZ and pcDNA3 GDNF could be transferred into retinal neural cells and the expressive product of GDNF gene, a 32 KD protein band, was detected. CONCLUSION: The preliminary results suggest that LacZ gene could be a marker to identify gene transfer into the target tissue and cells; GDNF gene could be transferred into cultured retinal neural cells of SD rats, and express 32 KD protein. PMID- 12580031 TI - [The expression of human leukocyte antigen from peripheral blood of patients with herpetic keratitis]. AB - PURPOSE: To understand the correlation between the expression of human leukocyte antigen from peripheral blood and herpetic keratitis so as to find the pathogenesis of HSK. METHODS: The expression of HLA-ABC, HLA-DR, HLA-DP, HLA-DQ from peripheral blood of 32 cases with herpes simplex keratitis (the stromatic and ulcer herpetic keratitis are 16 cases respectively), comparing with the normal people, were investigated by flow cytometry. RESULTS: The expression rate of HLA-ABC from peripheral blood of patients with herpetic keratitis (both the stromatic and ulcer groups) was lower, and the expression of HLA-DR and HLA-DP was higher than normal people. No difference was found between the stromatic and the ulcer herpetic keratitis in the expression of HLA-ABC, HLA-DR, HLA-DP, HLA DQ. CONCLUSION: The immune model in patients with herpetic keratitis is different from normal people. The individual with this kind of immune model can not destroy the infected virus effectively, also show autoimmunization to cornea. PMID- 12580032 TI - [Serum soluble interleukin-2 receptor levels in uveitis]. AB - PURPOSE: To investigate the relationship between serum SIL-2R levels and uveitis. METHODS: SIL-2R levels were measured in sera of 64 patients with active uveitis by ELISA assay, 61 healthy subjects as control. RESULTS: Serum SIL-2R levels in the patients with active uveitis were significantly higher than that in the controls, but there were no significant difference among anterior uveitis, intermediate uveitis, Behcet disease and Vogt-Koyanagi-Harada syndrome. After therapy the serum SIL-2R levels were obviously decreased. CONCLUSION: SIL-2R Levels in serum correlate with activity of uveitis. PMID- 12580033 TI - [The observation of proliferative cell nuclear antigen and AgNORs in ocular adnexal lymphoid hyperplasia]. AB - PURPOSE: To investigate the expression of proliferative cell nuclear antigen and Nuclear Organizer Region in ocular adnexal lymphoid hyperplasia and its histopathologic classification. METHOD: Using anti-proliferative cell nuclear antigen monoclonal antibody (PC10) and AgNORs method, the proliferative cell in 34 cases of ocular adnexal lymphoid neoplasms (benign lymphoid lesion, BLL, 14 cases; atypical lymphoid hyperplasias, AH, 9 cases; malignant lymphomas, ML, 11 cases) were detected in formalin fixed and paraffin-embeded tissue. RESULTS: A significant difference between BLL and AH, BLL and ML was observed (PCNA P < 0.001, P < 0.001; AgNORs P < 0.01, P < 0.001); similar result was observed between AH and ML in PCNA index (P < 0.01). But there was not a significant difference in AgNORs count (P > 0.05). The linear regression coefficient between PCNA index and AgNORs number was excellent (0.8693). CONCLUSION: The two methods may be helpful to distinguish ocular adnexal lymphoid reactive hyperplasia and malignant lymphoma. PMID- 12580034 TI - [Study on human eye ciliary muscule cell culture and biologic characteristics]. AB - PURPOSE: We cultured human ciliary muscle [HCM] cells to study their growth, ultrastructure, immunohistochemistry and functional characters. METHODS: HCM cells from 10 young donor eyes were cultured with collagenase IV digestion procedures in vitro, the cells were identified by eletronmicroscope and immunohistochemistry assay, their function were studied by single-cell contraction assay. RESULTS: The cells were passed and grew in Hill-Valley pattern after conflunet; abundant filaments were presented under electronmicroscope. In Desmin protein immunohistochemistry study, the cultured cells were stained positive; in tissue sections, HCM cells stained positive, vascular smooth muscle stained positive weakly, but fibroblast cells and endothelial cells stained negative. 10(-3) M Carbachol could induce the cultured cells contract, this effect was antagonized by 10(-3) M Atropine. CONCLUSION: We successfully cultured HCM cells, which were able to contract. PMID- 12580035 TI - [The study on the culture, freezing preservation and resuscitation of human trabecular cells in vitro]. AB - PURPOSE: To establish the culture of human trabecular cells in vitro and study the freezing preservation and resuscitation. METHODS: The trabecular specimens from human eyes were cultured in vitro and passaged 3 times, then the cultured cells were frozen and preserved. The frozen cells were resuscitated after one week, two weeks, one month and two months. RESULTS: Cultured human trabecular cells were obtained. The frozen cells were resuscitated successfully. All of the resuscitated rates were more than 90%. CONCLUSION: The human trabecular cells were cultured in vitro successfully, they can also be frozen and resuscitated, which will be helpful to the establishment of cDNA library and the screening of associated glaucomatous genes. PMID- 12580036 TI - [Precollagen III in the vitreous of eyes with proliferative vitreoretinopathy]. AB - PURPOSE: To detect precollagen III (PC III) levels in serum and vitreous samples from patients with proliferative vitreoretinopathy (PVR). METHODS: PC III levels were measured by radioimmunoassay in 5 cadaveric vitreous samples from normal subjects, 20 normal human serum samples, 29 vitreous and serum samples from patients with rhegmatogenous retinal detachment complicated with PVR. The relationship between PC III levels and PVR history, surgery, epiretinal membrane, severity of PVR were analyzed with Logistic regression analysis. RESULTS: The mean PC III levels in serum samples of PVR patients and control subjects were 83.76 +/- 18.52 and 85.02 +/- 17.50 micrograms/L respectively (P > 0.05). PC III was not detected (< 40 micrograms/L) in the cadaveric vitreous. In vitreous aspirates from 29 eyes with PVR, it was not detected in 12 eyes, but higher than 40 micrograms/L in 17 eyes with the mean level of 268.69 +/- 176.07 micrograms/L. (P < 0.05) The high levels correlated with duration and severity of PVR. As onset time and PVR grade increasing, the probability of PC III concentration higher than 40 micrograms/L was 5.2655 and 2.7978 times the concentration lower than 40 micrograms/L respectively. CONCLUSION: PC III can be detected in vitreous aspirates from PVR eyes. The PC III levels in the vitreous was an event of local responses and were correlated with severity and history of PVR. This suggests that PC III and collagen III be involved in the development of PVR. PMID- 12580037 TI - [The establishment, freezing-state preservation and resuscitation of retinoblastoma SO-Rb70 cell line]. AB - PURPOSE: To establish a new cell line of human retinoblastoma (Rb), frozen and resuscitated to provide a Rb cell reservoir for clinical and experimental research. METHODS: The screening cells were washed and blown directly without centrifugation and then inoculated. Based on the cell-frozen principle, the Rb cell line was collected and established, then frozen in liquid nitrogen and resuscitated. RESULTS: SO-Rb70 cell line was established successfully. 8 groups of cells were frozen. The resuscitation experiment was carried out for 6 times, 4 times being successful. CONCLUSION: The preserved human Rb cells in liquid nitrogen grew normally after resuscitation with the maintenance of previous bioactivities and characteristics. This research helped to provide abundant material for further study of Rb. PMID- 12580038 TI - [The real-time observation of anterior segment of primary congenital glaucoma]. AB - PURPOSE: To study the characteristics of primary congenital glaucoma in ultrasound biomicroscopy and discuss the pathogenic mechanism of primary congenital glaucoma. METHODS: Ten eyes of 6 congenital glaucoma patients and 7 eyes of 7 normal persons were examined using ultrasound biomicroscope (UBM). RESULTS: UBM can show the dysplasia of anterior angle, iris, ciliary body and scleral spur of primary congenital glaucoma. The base of iris is thin, the ciliary body is small, and the anatomic characteristic is not clear. The thickness of iris and size of ciliary body of primary congenital glaucoma are significantly smaller than those of normal control group. CONCLUSION: The results suggest UBM can inspect some part of eye where the normal examinations can not reach, e.g., ciliary body, the posterior surface of iris, etc. The characteristics of congenital glaucoma in UBM include thin iris, small ciliary body and unclear scleral spur. That suggests congenital glaucoma not only has dysplasia of anterior angle, but also has dysplasia of iris, ciliary body and scleral spur. The dysplasia of iris and ciliary body may play a relative important role in the pathogenic mechanism of congenital glaucoma. PMID- 12580039 TI - [Effect of AHMED valve implantation surgery for the treatment of neovascular glaucoma]. AB - PURPOSE: To evaluate the efficacy of the valve implantation for the treatment of neovascular glaucoma. METHODS: 15 cases (15 eyes) neovascular glaucoma were treated with AHMED valve implantation surgery, and followed up for 6 to 42 months. RESULTS: The IOP of the cases is lowered from 6.01 +/- 1.28 kPa (1 kPa = 7.5 mmHg) preoperatively to 1.96 +/- 0.68 kPa postoperatively. The success rate was 80.0% (The criteria of success is 0.80 kPa < or = IOP < or = 2.80 kPa and without severe complication that influence the visual acuity). The common postoperative complications included hyphema, shallow anterior chamber. CONCLUSION: The AHMED valve implantation surgery can lower the IOP, increase the operative success rate and preserve the visual function effectively. It is regarded to be an effective, safe method to treat neovascular glaucoma. PMID- 12580040 TI - [Management of very complicated retinal detachment]. AB - PURPOSE: To evaluate the management for a group of complicated retinal detachment in very severe conditions including(A) rhegmatogenous retinal detachment associated with severe posterior and anterior PVR, (B) giant retinal tear (GRT) retinal detachment with severe PVR, (C) severe perforating eye injury with tractional retinal detachment. METHODS: A total number of 31 eyes from 31 patients was included, in which 14 eyes were in group A, 8 eyes were group B and 9 eyes group C. Procedures including scleral buckling, encircling, cryotherapy, total vitrectomy, epiretinal membrane peeling/excision, relaxing retinectomy, use of perfluorocarbon liquid and silicone oil were applied to all patients. Patients were followed up for three to 16 months. RESULTS: Retinas of 21 out of 31 eyes(67.7%) were completely reattached and 5 eyes (16.1%) were partially reattached, 5 eyes were failed(16.1%). Postoperative vision was significantly improved in 25 eyes. The visual acuity of 19 eyes was better than 0.02, while before operation only two eyes were better than 0.02. PVR and GRT patients had better results. CONCLUSION: The results showed that a big part of very complicated retinal detachment which was called as hopeless cases could still be successfully treated with comprehensive procedures, more than half of them may have useful vision which may obviously improve their life quality. PMID- 12580041 TI - [The application of retinotomy and retinectomy for complicated retinal detachment]. AB - OBJECTIVE: To investigate the techniques, therapeutic effects, indications, complications and control of retinotomy and retinectomy for complicated retinal detachment. METHODS: Based on the preliminary report of retinotomy and retinectomy, 36 eyes of 36 patients of complicated retinal detachment were operated on and followed-up for 3-12 months. The operative technique included buckling, vitrectomy, peeling, retinotomy and retinectomy, intraocular tamponade and laser treatment. RESULTS: Retina was reattached in 32 eyes in operation. Retinal detachment was recurrent in 8 eyes in followed-up in which retina was reattached again in 3 eyes by second operation. The last successful rate was 75% (in 27 eyes). The visual acuity was worse than funger conter in 33 eyes and worse than 0.05 in 3 eyes before operation. The visual acuity was better than 0.05 in 20 eyes (55.5%) after operation. The complications included bleeding, retinotomy expanding, complicated cataract, hypotony and so on. CONCLUSION: Retinotomy and retinectomy can improve the curative effect of complicated retinal detachment. But the complications of these operation were severe. The indications should be well controlled. PMID- 12580042 TI - [Vitrectomy for the treatment of retinal detachment uncomplicated by advanced proliferative vistreoretinopathy]. AB - PURPOSE: To determine the role of vitrectomy be used in retinal detachment uncomplicated by proliferative vitreoretinopathy (PVR). METHODS: We studied 54 cases who had undergone vitrectomy for retinal detachment uncomplicated by severe proliferative vitreoretinopathy. The indications for vitrectomy fell into three main groups: 1) retineal detachment of PVR-B (n = 15), 2) Retinal breaks at the posterior of PVR-C1 or C2 (n = 11), 3) retinal view was poor (n = 28). RESULTS: The success rate with one procedure was 74% and with further surgery retinal reattachment was achieved in 96%. After surgery, the visual acuity was improved in 43 eyes (79.6%) unchanged in 9 eyes (16.7%), and worse in 2 eyes (3.7%). CONCLUSION: Vitrectomy is an effective method for treatment of selected cases of retinal detachment uncomplicated by proliferative vitreoretinopathy. PMID- 12580043 TI - [The relation of diabetic retinopathy with blood-fat and lipid peroxidation]. AB - PURPOSE: To observe and inquire into the relation of diabetic retinopathy with lipid metabolism, and lipid peroxidation. METHODS: The contents of triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), superoxide dismutase (SOD) and lipid peroxide (LPO) of serum were examined and determined both in thirty patients with diabetes in early stage suffering from retinopathy and twenty four healthy people. RESULTS: TG, TC, LDL-C and LPO in patients group were obviously higher than those in the control group, while HDL-C and SOD were lower than those in healthy control group. CONCLUSION: The disorder of lipid metabolism and the growing intensity of lipid peroxidation caused by diabetes may be one of the dangerous factors to bring about the retinopathy. PMID- 12580044 TI - [B-scan ultrasonic imaging of morning glory syndrome]. PMID- 12580045 TI - [Observation of complications at the operative and early postoperative stages of phacoemulsification]. AB - PURPOSE: The study was designed to investigate the reason and management of the complications at the initial stage of studying the technique of phacoemulsification. METHODS: Observing the complications at the operative and early postoperative stages of phacoemulsification used in situ nuclear fracture technique or in iridial plain fracture technique on 233 eyes of 212 patients with cataract. RESULTS: During the operations the rate of vitreous loss was 8.6% (20 eyes); iridial damage was 5.6% (13 eyes); hemorrhage in the anterior chamber was 0.4% (1 eye). After the operations the rate of continued corneal edema was 1.3% (3 eyes); posterior capsular opacity was 3.0% (7 eyes); inflammation was 0.9% (2 eyes); discentral location of intraocular lens was 0.4% (1 eye); leakage of wound was 0.9% (2 eyes); hemorrhage in the anterior chamber was 0.4% (1 eye). CONCLUSION: Vitreous loss and iridial damage were the frequent complications at the initial stage of studying the technique of phacoemusification. The rate of the complications was lower in situ nuclear fracture technique than in iridial plain fracture technique. PMID- 12580046 TI - [The application of antisense technology in ophthalmology]. AB - The research of gene regulation become more and more important with the advancement of the technology of molecular biology and gene study. Antisense molecules can regulate and modify gene expression. By using antisense technology, antisense RNA or artificial antisense oligonucleotides can be made to inhibit or block the transcription and translation of genes. These molecules are highly specific in gene regulation. The application of antisense technology in ophthalmology is introduced briefly in the review. This may be helpful to the studies in ophthalmic research field. PMID- 12580047 TI - [Optical coherence tomography and its application in ophthalmology]. AB - Optical coherence tomography (OCT) is a new imaging modality that provides cross sectional tomographs of ocular structure in vivo. OCT system utilizes propagation of light through, and the time-of-flight delay of light reflected back from biological tissue to produce false-color images of microstructure. OCT has superior spatial resolution (10 microns) than other optical imaging instruments, and maybe achieved at arbitrary penetration depth in transparent tissue with a noncontact, noninvasive measurement. Such features make OCT useful for the early diagnosis, differential diagnosis and more sensitive monitoring of a variety of ocular abnormalities including glaucoma, retinal and anterior eye diseases. PMID- 12580048 TI - [Effects of endomorphins and their analogs on cardiovascular system]. AB - AIM: To analyze the effects of endomorphins (EMs) and their analogs ([D-Pro2]EM 1, [D-Ala2]EM-1, [D-Pro2]EM-2 and [D-Ala2]EM-2) on the cardiovascular system of anestetized rats and to study its mechanism. METHODS: Responses to EMs and their analogs were investigated in the systemic vascular bed of rats and the mesenteric microarteria of Bufo gargarizans. Responses to EMs were investigated on the hindquarters of the rat vascular bed under constant-flow conditions and on the isolated rabbit thoracic aorta strips. RESULTS: The EMs and their analogs showed dose-related (10(-9)-10(-6) mol.L-1, i.v.) and naloxone-sensitive (2 mg.kg-1, i.v.) hypotension in mean arterial pressure of rats with similar duration and vasodilatation in mesenteric microarteria of Bufa gargarizans. The sequence of potencies was EMs > [D-Pro2] EM-2 > [D-Ala2]EM-2 > [D-Ala2]EM-1 > [D-Pro2]EM-1. EMs were shown not to relax the isolated rabbit thoracic aorta strips without endothelium. EMs, however, relaxed them with endothelium significantly. This action was blocked by Nx (10(-5) mol.L-1) and L-NNA (10(-4) mol.L-1). CONCLUSION: The significant hypotensive activity of EMs and their analogs is mainly associated with their vasodilatation, which is related to the release of NO from vascular endothelium, and their potency is not completely related to their affinity for mu-opiate receptor. PMID- 12580049 TI - [Effect of lidamycin on the expression of genes involved in invasion regulation in HCT-8 human colon cancer cells]. AB - AIM: To study the effect of lidamycin on the expression of genes involved in invasion regulation in HCT-8 human colon cancer cells. METHODS: HCT-8 human colon cancer cells were treated with lidamycin (10 nmol.L-1) for 8 h. The effect of lidamycin on the expression of genes were detected by cDNA arrays. Northern blot and RT-PCR. RESULTS: Hybridization of the entire cDNA populations to Atlas Arrays showed that lidamycin down-regulated the expression level of MMP-9 and up regulated the expression level of TIMP-1. These changes were confirmed by Northern blot and RT-PCR. CONCLUSION: The results indicate that lidamycin may exhibit its anti-invasive activity by inhibitting the production of type IV collagenase whilst enhancing the production of tissue inhibitor metalloproteinase. PMID- 12580050 TI - [Effects of 4-aminopyridine on calcium currents and sodium currents in guinea pig ventricular myocytes]. AB - AIM: To investigate the effect of 4-aminopyridine (4-AP) on ion channels of myocytes. METHODS: L-type calcium channel and sodium channel currents were recorded in guinea pig single ventricular myocyte using whole-cell patch-clamp techniques. RESULTS: 4-AP, 0.1, 0.5 and 1.0 mmol.L-1 were shown to inhibit L-type calcium channel currents (ICa, L) and sodium channel currents (INa) concentration dependently. The percentage of inhibition were (11.6 +/- 1.7)%, (37.5 +/- 8.3)% and (54.5 +/- 6.9)% (P < 0.01) respectively for ICa, L, and (22.1 +/- 14.3)% (P < 0.05), (39.4 +/- 8.8)% and (62.3 +/- 6.8)% (P < 0.01) respectively for INa. 4-AP 0.5 mmol.L-1 shifted the I-V curves of ICa, L and INa upwardly. CONCLUSION: 4-AP blocked L-type calcium channel and sodium channels in guinea-pig ventricular myocytes concentration-dependently. PMID- 12580051 TI - [Effects of artesunate on progestrone estrogen content and decidua in rats]. AB - AIM: To observe the effect of artesunate on content of progesterone, estrogen and decidua of pregnant rats and study the efficacy and mechanism of artesunate for termination of early pregnancy. METHODS: Serum content of progesterone, estrogen and TNF-alpha were measured with RIA. The effects of artesunate on the ovary, decidua and fetus of pregnant rats were studied using histochemistry techniques. Decidual cells were estimated using cell culture. RESULTS: Artesunate 40 mg.kg-1 s.c. on day 6-10 of gestation significantly decreased the concentration of serum progesterone in early pregnant rats; decidual cells and fetus of treated groups were found to be degenerated at d 11. Artesunate was shown to directly damage the decidual cells. Cultured human decidual cells were exposed to artesunate for 48 h, the LC50 was found to be 25 +/- 3 mL.L-1. CONCLUSION: The damage of artesunate on decidua and placenta may be the mechanisms of its contragestational action. PMID- 12580052 TI - [5,6-diaryl-2,3-dihydro-1-pyrrolizinone derivatives synthesis and antiiflammatory and analgesic activities]. AB - AIM: To search for more potent and less toxic antiinflammatory and analgesic activity compounds. METHODS: A series of 5,6-diaryl-2,3-dihydro-1-pyrrolizinone derivatives were designed and synthesized based on the structures of diarylheterocyclic COX-2 selective inhibitors. Their structures were determined on the basis of spectal data (IR, MS and 1HNMR). Their antiinflammatory and analgesic activities in vivo were tested by xylene-induced mouse ear edema model and acetic acid-induced mouse writhing model p.o. dose of 200 mg.kg-1. RESULTS: Seventeen new compounds (1-17) were synthesized. Many of these compounds showed antiinflammatory and analgesic activities. CONCLUSION: Compound 3, 8, 11, 14 and 15 showed antiinflammatory activities more potent than ibuprofen. Compound 9, 10 and 11 showed analgesic activities comparable to ibuprofen. These compounds are regarded to be promising to develop new potent drugs. PMID- 12580053 TI - [Study on 3D-QSAR of PPAR gamma agonists with thiazolidinedione and arylketo-acid moieties]. AB - AIM: To build a model of two series of PPAR gamma agonists--thiazolidinedione and aryketo-acid derivatives using 3D-QSAR method, and to reveal the structural features affecting the binding activity to PPAR gamma, which relates to antihyperglycemic and antihyperlipidemic activity and has a potential application to the treatment of type II diabetes. METHODS AND RESULTS: 48 agonists with selective activity for PPAR gamma were analyzed using CoMFA. Based upon the active conformation of rosiglitazone (BRL) extracted from its complex with PPAR gamma all agonists were aligned. The model from CoMFA showed a high ability to explain and predict the activity of PPAR gamma agonists with cross-validation correlation coefficient R2 = 0.656, that of non-cross-validation R2 = 0.982, F10,37 = 201.1, and SE = 0.115. CONCLUSION: The CoMFA contour map indicates that the steric fields mainly contribute to the binding effect, and especially a bulk group in the arylketo-acid series favors in the increase of affinity for PPAR gamma, as compared to the thiazolidinedione. PMID- 12580054 TI - [Synthesis of substituted 4-styrylcoumarin and their antitumor activities]. AB - AIM: A series of 4-styrylcoumarin derivatives had been designed and synthesized in order to find compounds of antitumor activities by screening. METHODS: Title compounds (1-20) were synthesized by Phase-Transfer Wittig-Horner reaction, and screened by several antitumor models in vitro. Their structures were determined by 1HNMR, MS and elemental analysis. RESULTS: Twenty compounds (1-20) are new compounds. Compound 18 had effects on KB cell lines in vitro. CONCLUSION: It was seen that compound 18 had certain antitumor activities, and it was worth further studying. PMID- 12580055 TI - [Synthesis and biological activities of new 5-HT2A selective ligands--N substituted-piperidinyl-4-phenylthioether and sulfone derivatives]. AB - AIM: A series of 4-piperidinylthioether and sulfone derivatives of 4-[1-hydroxy-1 (2,3-dimethoxyphenyl) methyl]-N-2-(4-fluorophenylethyl) piperidine (MDL 100907) were synthesized in order to find new 5-HT2A selective ligands. METHODS: Title compounds 2a-2c were synthesized from 2, 3-dimethoxythiophenol and tested for their affinities to 5-HT2A, 5-HT2C, 5-HT6 and 5-HT7 receptors and some other nervous transmitter receptors in vitro. RESULTS: Compounds 2a-2c are new compounds. The results of the binding assay demonstrated that they have relatively high selectivity for 5-HT2A receptor in vitro. CONCLUSION: Some sulfur containing analogues of MDL 100907 showed selective affinity to 5-HT2A receptor and are worth further study. PMID- 12580056 TI - [Studies on the chemical components of Clematis chinensis]. AB - AIM: To study the chemical components of the root of Clematis chinensis. METHODS: Various chromatographic techniques were used to separate and purify the components. Their structures were determined on the basis of spectral and chemical evidence. RESULTS: Two compounds were isolated and elucidated as: clemaphenol A (I) and dihydro-4-hydroxy-5-hyroxymethy-2(3H)- furanone (II). CONCLUSION: Compound I and II are new compounds. PMID- 12580057 TI - [Studies on the chemical constituents of the seeds from Artabostrys hexapetalus (Annonaceae)]. AB - AIM: To study the chemical constituents of the seeds from Artabotrys hexapetalus (L.f.) Bhandari (Annonaceae). METHODS: Various chromatographic techniques were used to separate and purify the constituents. Their structures were elucidated on the physico-chemical properties and spectral data. RESULTS: Eight compounds were isolated from the seeds of A. hexapetalus. They were identified as four neolignans: isoamericanin A (1), isoamericanol A (2), americanin B (3) and artabotrycinol (4), a semiterpenoid: (R)-artabotriol (5) and others: palmitic acid (6), beta-sitosterol (7) and daucosterol (8). CONCLUSION: Artabotrycinol (4) and (R)-artabotriol (5) are new compounds. Three other neolignans were isolated from this plant for the first time. PMID- 12580058 TI - [Cycloartane triterpenes and glycosides from Cimicifuga acerina]. AB - AIM: To investigate the chemical constituents from the rhizomes of Cimicifuga acerina (Sieb. et Zucc.) Tanaka. METHODS: Column chromatgraphy (including silica gel and ODS) together with HPLC was used to separate the chemical constituents whose structures were determined by FAB-MS, NMR (1D and 2D) and hydrolysis methods. RESULTS: Five cycloartane triterpenoids were isolated and identified as: (22R)-22-hydroxycimigenol (I), (22R)-22-hydroxy-24-O-acetylhydroshengmanol 3-O beta-D-xylopyranoside (II), dahurinol (III), 24-epi-24-O-acetyl-7,8 didehydroshengmanol 3-O-beta-D-xylopyranoside (IV), 25-O-acetyl-7,8 didehydrocimigenol 3-O-beta-D-xylopyranoside (V). CONCLUSION: Compound I is a new natural product, compound II is a new compound and compounds IV and V were isolated from this plant for the first time. PMID- 12580059 TI - [Analysis of tannins in Fructus Chebulae and its confusion varieties by HPCE]. AB - AIM: To analyze the hydrolyzable tannins-chebulinic acid (I) and chebulagic acid(II) in Fructus Chebulae and its confusion varieties by using high performance capillary electrophoresis (HPCE) method. METHODS: Using a capillary (375 microns OD x 50 microns ID; 81.5 cm x 61.5 cm) and a power supply set at 24 kV, with phosphate-borate buffer containing 20 mmol.L-1 Na2HPO4-60 mmol.L-1 boric acid and a UV detector at 280 nm, sample solution was loaded in decompression mode at the positive end of the capillary, the loading time was 5 s. RESULTS: The linear ranges of I and II were 0.0842-0.842 and 0.842 and 0.0940-0.940 mg.mL-1 respectively, the correlation coefficient were 0.9966 and 0.9957, the average recoveries were 95.6% (RSD = 4.0%, n = 5) and 95.0% (RSD = 4.4%, n = 5), the RSDs (n = 5) of measurement precision test were 2.2% and 1.7%, the RSDs (n = 6) of reproduction test were 5.4% and 4.0% respectively. The contents of I and II were obviously interrelated with the variety and characteristics of Fructus Chebulae, the contents of I and II in the confusion varieties of Fructus Chebulae were very low. CONCLUSION: It is suitable to use I and II as the criterion in quality evaluation of Fructus Chebulae, and the HPCE method is effective for quality evaluation of the crude Fructus Chebulae. PMID- 12580060 TI - [Adsorptive voltammetric behavior of omeprazole and its application]. AB - AIM: To study the electrochemical behavior of omeprazole (OPZ) and a new method for the determination of the compound was established by adsorptive stripping voltammetry. METHODS: Linear-sweep polarography and cyclic voltammetry were used. RESULTS: In a supporting electrolyte containing 0.1 mol.L-1 NH3-NH4Cl, a reduction peak of OPZ was observed by linear-sweep voltammetry at Hg electrode. The peak showed a potential of -1.04 V (vs Ag/AgCl at 100 mV.s-1) and adsorptive characteristics. The adsorbed species is most probably the neutral molecular of OPZ. The saturated adsorption of OPZ at Hg electrode is 9.55 x 10(-11) mol.cm-2 and every OPZ molecule occupies and area of 1.74 nm2. On the surface of the hanging mercury drop electrode, the adsorption of OPZ obeys Frumkin adsorption isotherm. The adsorption coefficient beta is 1.32 x 10(6), the Gibbs energy of adsorption delta G theta is -34.93 kJ.mol-1, the number of electrons transferred n is 4, the kinetics parameter alpha n alpha is 0.78, the rate constant of the surface electrode reaction ks is 0.18 s-1, the diffusion coefficient D of OPZ in solution is 1.03 x 10(-5) cm2.s-1. The detection limit of the adsorptive stripping voltammetry of OPZ is 2.0 x 10(-9) mol.L-1 under optimized condition. CONCLUSION: This indicates an irreversible process with adsorption. PMID- 12580061 TI - [Determination of imperialine and imperialine-3 beta-D-glucoside in bulbs of Fritillaria pallidiflora by HPLC-ELSD]. AB - AIM: To establish an HPLC (high performance liquid chromatography)-ELSD (evaporative light scattering detector) method for the quantification of chromophore-lacking imperialine (I) and imperialine-3 beta-D-glucoside (II) in bulbs of Fritillaria pallidiflora. METHODS: By optimizing the chromatographic conditions of HPLC and the parameters of ELSD to test the methodology. Column: Kromasil C-18 (5 microns, 200 mm x 4.6 mm), mobile phase: acetonitrile-water triethylamine (71:29:0.03), flow rate: 1.0 mL.min-1, drift tube temperature: 88 degrees C, gas flow-rate: 2.38 L.min-1. RESULTS: The calibration curves were linear in the range of 60.72-161.92 micrograms.mL-1 for I and 48.72-194.88 micrograms.mL-1 for II. The precisions (RSD) of intra-day and inter-day were less than 1% for I and II. The recoveries of the amounts of 24.28, 30.35 and 36.42 micrograms for I were 97.1%, 96.6% and 96.8%, and 24.20, 30.24 and 36.29 micrograms for II were 95.8%, 96.9% and 96.0%. The minimal detectable concentrations of I and II were 2.43 and 1.53 micrograms.mL-1 respectively. CONCLUSION: The established method is simple, rapid, accurate and sensitive, and can be utilized to determine the contents of imperialine and imperialine-3 beta-D glucoside in crude drug Yibeimu (derived from the bulbs of Fritillaria pallidiflora). PMID- 12580062 TI - [Studies on lung targeted niosomes of carboplatin]. AB - AIM: To improve the treatment efficacy and reduce the side effect of carboplatin (CBP), the antitumor drug was incorporated into niosomes (NS). METHODS: Lung targeted niosomes of CBP (CBP-NS) were prepared by the method of hand shaking. The ultraviolet absorption spectrophotometric method was used for the determination of CBP. The method of dynamic dialysis was used for in vitro release of CBP from CBP-NS. The S-180 lung neoplasms models were established by i.v. cancer cells in mice. The number of pulmonary nodules was examined for evaluating the therapeutic efficacy. RESULTS: The data showed that the mean diameter of CBP-NS was 3.72 microns, with a span of 0.66. The entrapment ratio of CBP in CBP-NS was 29.2% and the CBP-NS were stable for three months stored at 3 degrees C-5 degrees C, 15 degrees C-25 degrees C or 37 degrees C (relative humidity 75%). The release profile in vitro could be described by a biexpotential equation. The calculated values of the three targeting parameters indicated that CBP-NS showed good targeting efficiency. The results of therapeutic trials showed that the antitumor effects were significantly increased by injection of CBP-NS compared with CBP in the treatment of mice with lung carcinoma. CONCLUSION: The results indicated that CBP-NS have good targeting efficiency in vivo, and the biodegradable CBP-NS may decrease the side effects of carboplatin and improve its therapeutic efficacy. PMID- 12580063 TI - [Effect of 4-aminopyridine on taurine-regulated release of amino acid from rat cortical synaptosomes]. AB - AIM: To elucidate the mechanism of taurine-regulated amino acid release from synaptosomes. METHODS: Endogenous aspartate, glutamate and GABA release from cortical synaptosomes were measured by high performance liquid chromatography using stepwise elution system, Glutamate release was monitored by continuous fluorometry. RESULTS: 4-Aminopyridine (3.0 x 10(-2) mol.L-1) counteracted the taurine-induced inhibition of glutamate overflow (P < 0.05), while aspartate and GABA release was not affected. Nimodipine (10(-5) mol.L-1) combined with 4 aminopyridine was shown to decrease glutamate release (P < 0.05). CONCLUSION: Taurine may regulate glutamate release through presynaptic L-type calcium channel and also act on Asp- and GABA-nereve terminal to regulate Asp and GABA release in rat cortex. PMID- 12580064 TI - [Synthesis and antibacterial activities of 3,6-disubstituted-5,6-dihydrogen-1,2,4 s-triazolo-[3,4-b]-1,3,4-thiadiazoles]. AB - AIM: To synthesize a series of new fused heterocyclic compounds containing 1,2,4 triazole and 1,3,4-thiadiazole rings. METHODS AND RESULTS: Nine new fused heterocyclic compounds have been synthesized by the reaction of 3-p nitrophenoxymethyl-4-amino-5-mercapto-1, 2, 4-triazole with aromatic aldehydes in the presence of acid. The reaction conditions for the synthesis have been investigated. The structures of the synthesized compounds were confirmed by elemental analysis, IR, 1HNMR and MS. All of the compounds were screened for their biological activities. CONCLUSION: The results show that some compounds (2c, 2d, 2e, 2f) showed strong antibacterial activities. PMID- 12580065 TI - [Structural elucidation of a new perylenequinone]. AB - AIM: To study the chemical constituents from the mycelia of Hypomyces sp. METHODS: Silica gel column chromatography was employed for the isolation and purification. The structure of compound 1 was elucidated on the basis of spectral analysis. RESULTS AND CONCLUSION: A new perylenequinone, named hypomycin B 1, was isolated from the mycelia of Hypomyces sp. PMID- 12580066 TI - [The development of electrospray ionization mass spectrometry (ESI-MS) in studies of non-covalent protein complexes]. PMID- 12580067 TI - Dacryology in China. AB - PURPOSE: International Society of Dacryology held the IV International Congress in Stockholm on June 1996. I was warmly invited as a guest of honor beforehand to deliver a lecture on a special topic of "Dacryology in China" there, so that the world might understand the dacryology in China, and thus the international growth of dacryology as a discipline would be promoted. METHODS: In China tear lab research and clinical research of LG are nearly empty except a few papers published. We had studied 3 items on LS for 28 years before 1979, and put our attention mainly on dacryology since 1979 by 3 stages: (1) collecting literatures (1979-81); (2) lab research (1981-84) and (3) clinical research, put DE as the key point by 4 steps: DE diagnosis (1984-86); DE treatment (1987-91); DE Condition Judgement (1989-92) & DE cause & mechanism (1991-95). RESULTS: In the past 44 years, esp. from 1979 on, in a planned way, we have systematically researched the dacryology with main results below: created new word "Leiyexue" in Chinese & its English words, dacryology & lacrimology by adding-ology to dacrya & lacrima; made LS consisted of reflex LG & LP, and basic LG besides; found out the normal & abnormal data of tear and serum LF & its iron bindingcapacity; MDH, LDH & isoenzymes; beta 2 m; MDA & antioxidants; trace elements, prostaglandins, intramuscular gentamicin passing the blood-tear barrier; bacteria, fungi, anaerobe, virus & parasite; Ig, T cell & subsets, Langerhan cell, HLA antigen; in cidence (KCS, 0.3-3%, trancomatous DE 0.003%); the parallel correlations between DE graded by rb, BUT & Slt; and in SS, lymphocytes infiltration in LG; Lab G & conjunctiva; LF (< 100 mg%) & beta 2 m (> 1.7 mg%); and testosterone receptors in LG; negated presence of tear Beta-lysim; set up a series of models for researching & a complete set of measures to diagnose, grade, judge and treat DE, deviced new operations. AT & prescription for DE; proved SS being autoimmune & EB virus, its one cause. CONCLUSIONS: This relatively, overall & systematical research of dacryology already have been widely applied to practice with over 100 papers published. It indicates that LS & its disease as a scientific discipline under the term of dacryology to grow up is entirely possible and extremely needful. PMID- 12580068 TI - An experimental study of the tumour necrosis factor level in aqueous humor after transscleral fixation of intraocular lens. AB - PURPOSE: The tumour necrosis factor (TNF) level in aqueous humor after transscleral fixation of intraocular lens (IOL) implantation in rabbits and discuss the effect of TNF on postoperative anterior ocular inflammation. METHODS: Twenty-seven pigmented rabbits were divided into three groups. Group 1: transscleral fixation of posterior chamber (PC) IOL implantation; Group 2: Lens of rabbits was removed without IOL implantation; Group 3: the control group, without surgical intervention. On the 1st, 3rd, 7th and 14th postoperative days, aqueous humor samples were obtained. An modified double antibodies indirect sandwich ELISA was used to detected for the presence of TNF. The data were analyzed by using analysis of variance of SAS software. RESULTS: It was found that TNF level in aqueous humor was increased after transscleral fixation of IOL implantation. TNF level reached its maximum on the 14th postoperative day in the IOL implanted group. TNF level on 1st, 3rd, 7th and 14th days postoperatively was significantly the highest (p < 0.05) in IOL implanted group when compared with the other two groups. CONCLUSIONS: There is something physically related to the IOL itself which induces the secretion of TNF or this simply related to mechanical factors related to the presence of the IOL itself, and the manipulation of the iris during lens implantation stimulates TNF secretion. The increase of TNF level may play critical role in anterior ocular inflammation response after IOL implantation. PMID- 12580069 TI - Determination of intraocular lens tilt and decentration using simple and rapid method. AB - PURPOSE: To determine the tilt and decentration of implanted intraocular lens (IOL) in vivo one year postoperatively. METHODS: A retrospective study was performed on 67 eyes by the method introduced by Guyton using the third and fourth Purkinje imagesm through undilated pupil. RESULTS: The average decentration of implanted IOL was 0.68 mm, and the average tilt was 6.03 degrees. The result was similar to the histopathological study in postmortem eyes and the measurement in vivo. Linear-regression analysis showed there was no relationship between the tilt and decentration and the residual astigmatism, axial length, IOL power, etc. CONCLUSION: The result is similar to the other methods. The method using the third and fourth Purkinje images is simple and easy to perform. It can be used for determining the IOL position through undilated pupil and does not require special apparatus. PMID- 12580070 TI - Effect of cataract surgery on ocular axial length elongation in young children. AB - PURPOSE: To evaluate the influence of cataract surgery on eyes of children between 1-5 years old. MATERIALS AND METHODS: Forty-seven cases of pediatric cataract, including 31 cases of bilateral congenital cataract and 16 cases of unilateral posttraumatic cataract, were retrospected. Of the 31 cases of congenital cataract, 19 cases (group 1) had undergone cataract extraction with intraocular lens (IOL) implantation, 12 (group 2) had undergone cataract extraction without IOL implantation. Each of them had their two eyes operated separately at an interval of 1 year or longer. Of the 16 cases of posttraumatic cataract, 6 (group 3) had undergone cataract extraction with IOL implantation and 10 (group 4) had undergone cataract extraction without IOL implantation. Mean age at the time of interested surgery was 2.9 years (range from 1.1 to 5.1 years). Ocular axial length of both eyes measured by A-scan ultrasonography before and after the surgery of the earlier operated eye in the congenital groups and the operated eye in the post-traumatic groups was recorded. The elongation of axial length of the earlier operated eye in the congenital groups and the operated eyes in the posttraumatic groups of each case were compared with that of the fellow eye statistically with paired t-test. RESULTS: The mean follow-up was 2.6 years (ranged from 1.0 to 6.4 years). In all the 4 groups, the elongation of axial length between the earlier operated eye and later operated eye in the congenital groups and between the operated and unoperated eyes in the posttraumatic groups of each case was not significantly different (p > 0.05). CONCLUSION: Cataract surgery, either extraction with or without IOL implantation, did not retard axial elongation in children above 1 year old. PMID- 12580071 TI - The multi-channel VEP topographies in patients with late-stage glaucoma. AB - PURPOSE: To study the clinical application of multi-channel VEP topographies in patients with late-stage glaucoma. METHODS: The multi-channel checkerboard reversal VEP waves of 25 normal persons and 15 patients with late-stage glaucoma were recorded and analyzed, All patients were examined using Humphrey Field Analyzer. The VEP topographies of multi-channel VEP waves were shown by computer processing system. RESULTS: In normal subjects, the topographies showed symmetric distribution to full-field pattern stimulation. In all patients with late-stage glaucoma, even whose visual field was severely damaged, multi-channel VEP could be recorded, All channels showed simple 'NPN' wave form, N1, N2 waves were not conspicuous, even both were lost, all channels showed the reduced amplitudes of P1 waves, and longer latency. The multi-channel VEP topographies showed irregular distribution, Some showed amplitude asymmetry, other limited or widespread, lateralized. CONCLUSIONS: Despite the severely damaged visual field, the patients with late-stage glaucoma showed special multi-channel VEPs topographies, Multi channel VEP topographies should be useful in following up patients suffering from late-stage glaucoma and studying visual processing in human brain. PMID- 12580072 TI - Electroretinographic oscillatory potentials in different aged rats: an analysis in the domains of time and frequency. AB - OBJECTIVES: Electroretinographic Oscillatory Potentials (Ops) are considered to be the optimal parameter to study the retinal circulation disturbances the present we compared parameters of OPs between different aged rats under dark adaptation. METHODS: The scotopic ERG were performed in anaesthetized rats aged 3, 12, 24 months. According a window of collective power spectrum field of the OPs was isolated by using the FFT and IFFT of the spectrum analysis program. The parameters in the time and frequency domains were calculated. RESULTS: Results showed that the amplitudes of wavelets and the summed amplitudes of OPs declined with the increase of age. A reduction of the total power and dominated power of OPs was also found after the FFT. But implicit time of each OP and dominant frequency did not change obviously. CONCLUSIONS: The present study indicated that change of OPs of rat might be related to senile degeneration of the retina and circulative deficiency in retina in elder rats. PMID- 12580073 TI - The study of local electrophysiology in macular diseases. AB - PURPOSES: To observe the features and clinical utility values of local electroretinogram (LERG) and local visual evoked potentials (LVEP) in macular diseases. METHODS: The spot stimuli were obtained by changing the shutters with different sizes of hole at the posterior of Ganzfeld ball. The recording electrode for LERG was made with the filament of platinum as blepharophthalmostat like. The LERG and LVEP were simultaneously recorded in macular diseases. RESULTS: The exudative AMD, Stargardt's disease, central serous chorioretinopathy and idiopathic macular hole had seriously decreased amplitudes of LERG and higher abnormal rates, comparing with atrophic AMD and idiopathic preretinal membranes in the region of 2.5 degrees and 5 degrees at macula. The abnormal rates in these diseases were higher for LERG than for LVEP. CONCLUSION: The LERG can objectively measure the variation of macular function in maculopathies. The sensitivities of LVEP are lower than those of LERG for macular diseases. PMID- 12580074 TI - The immunological studies on proliferative intraocular disorders: investigation of activated lymphocytes, macrophages and HLA-DR. AB - PURPOSE: To investigate whether activated B lymphocytes (CD23), activated T lymphocytes (CD25), macrophages (CD68) and human leucocyte antigen class II antigen (HLA-DR) were existed in epiretinal membranes (ERMs) and subretinal membranes (SRMs) of proliferative intraocular disorders (PID). METHODS: Twenty specimens of ERMs from rhegmatogenous retinal detachment with proliferative vitreoretinopathy (PVR), traumatic PVR and secondary traction retinal detachment, and two specimens of SRMs from rhegmatogenous retinal detachment with PVR and traumatic PRV were studied using immunohistochemical staining. RESULTS: CD68 and HLA-DR were found in all specimens, CD23 and CD25 in 4 cases of ERMs and in 1 case of SRMs, respectively. CONCLUSIONS: 1. The ERMs and SRMs of different etiology shared a common basis of inflammation and immunopathology. 2. There would be secondary cellular and humoral immunity in the ERMs and the SRMs of PID. PMID- 12580075 TI - Inhibition effect of cyclosporin A on human retinal pigment epithelial cells in vitro. AB - PURPOSE: To select effective drugs against cellular proliferation in the vitreous. METHODS: Cyclosporin A (0.125 mg/l-4.0 mg/l) was added to cultures of human retinal pigment epithelial (RPE) cells with or without macrophage conditioned medium (MCM). Proliferation rate of the cells was measured with (3H) thymidine incorporation and liquid scintillation techniques on days 3 and 5. RESULTS: Cyclosporin A at a dosages of 0.125 mg/l had a slight inhibition on human RPE cells proliferation (-3.8%-4.1%, P > 0.05). Cyclosporin A at a dose ranging from 0.25 mg/l to 4.0 mg/l inhibited cellular proliferation effectively and in a dose-dependent manner (8.7%-95.1%, P < 0.05 r = 0.94, P < 0.001) with its ID50 of 1.49 mg/l. In the culture of RPE with MCM, the inhibition on day 5 was more effective than that on day 3. CONCLUSION: Cyclosporin A had an effective inhibition on human RPE cell proliferation and it may be of potential use clinically. PMID- 12580076 TI - Cytotoxicity of contact lens care solution to human corneal epithelium in vitro. AB - PURPOSE: To investigate the cytotoxicity of soft contact lens multi-purpose care solutions which are now in common use in China. METHODS: The cell culture method was used. Cytotoxicity was indicated by significant increases in the number of dead cells relative to controls. RESULTS: Cells were exposed to soft contact lens care solutions for 15 min. They were irregular in shape and variable in size. The intercellular space increased and variable in size. The intercellular space increased and the cells became scrunken. With the time of exposure elongated, damage of cells became more severe. CONCLUSIONS: Four kinds of soft contact lens multi-purpose care solutions may have harmful effects on the culture of human corneal epithelial cells. Soaked lenses should be rinsed with saline before being placed in the eyes in order to reduce the potential toxicity of contact lens care solutions. PMID- 12580077 TI - An analysis of 1,001 blinding patients with corneal disease in 1960-1989. AB - PURPOSE: To investigate the condition and change of corneal blindness (CB) in the past 30 years. METHODS: 1,001 blinding patients of corneal disease were clinically analysed from 1960 to 1989, including etiology, sex, age, occupation, difference between the urban and rural areas etc. RESULTS: The male predominated. Infection stood the first cause of blinding corneal diseases, followed by trauma, malnutrition and the others. HSV-1 keratitis was the most frequent infection in 1980s. The highest incidence of CB was 20-40 year-old. The number of CB caused by occupational trauma was more than that by ordinary trauma in 1960s, but from 1970s, the latter was more than the former. The number of CB in rural area was more than that in the urban in 1960s, but from 1970s, there was no significant difference between the two areas. The percentage of bilateral CB was gradually decreased. CONCLUSION: We should try to find more effective strategies to prevent and treat CB caused by HSV-1 keratitis, pay special attention to preschool blindness caused by perforation injuries and avoid infection after corneal injuries of foreign bodies. PMID- 12580078 TI - Rieger syndrome. AB - PURPOSE: To discuss the clinic features of Rieger syndrome, the reasons of making wrong diagnosis, the way of treatment, and the research progress of its molecular characterization and gene mapping of this syndrome. METHODS: Two cases of Rieger syndrome which affected a patient and his daughter were studied. Multiple clinical examinations including photography of anterior segment, gonioscopy and fundus, Humphrey perimetry, A-scan ultrasonography, multiple tonometry in a day and chromosome examination were performed. Most importantly, ultrasonic biomicroscope (UBM) was first used to show the abnormalities of anterior segment in this syndrome. RESULTS: Gonioscopic examination revealed many mesoderm tissues remained and some parts of the iris adhered to cornea. In addition to cornea, iris and chamber angle, UBM showed that there was also hypoplasia of ciliary body. The result of the chromosome examination indicated normal. CONCLUSIONS: Rieger syndrome is an autosomal-dominated disorder with mesoderm dysgenesis. Recent researches have confirmed that a locus for this syndrome maps to 4q25. Besides hypoplasia of cornea, iris and chamber angle, its ocular phenotype maybe include dysgenesis of ciliary body which is probably one of the reasons of secondary glaucoma. PMID- 12580079 TI - [Inhibitory effects of indomethacin and meloxicam on NF-kappa B in mouse peritoneal macrophages]. AB - AIM: To study the inhibitory effects of indomethacin and meloxicam on NF-kappa B from lipopolysaccharide (LPS) stimulated peritoneal macrophages of mice. METHODS: NF-kappa B was measured with the method of electrophoretic mobility shift assay (EMSA). RESULTS: After induction by LPS at the concentrations of 1 and 3 micrograms.mL-1, the NF-kappa B content of the mouse peritoneal macrophages increased markedly. Indomethacin and meloxicam, at the concentrations of 10(-7) 10(-5) mol.L-1, decreased the activation of NF-kappa B at the concentrations of 1 and 3 micrograms.mL-1 in activated mouse peritoneal macrophages induced with LPS at the concentrations of 1 and 3 micrograms.mL-1. CONCLUSION: The inhibitory effects of indomethacin and meloxicam on NF-kappa B activation may be one of their mechanisms of antiinflammatory actions. PMID- 12580080 TI - [Effects of praeruptorin C on vascular hypertrophy, [Ca2+]i, collagen content and NO in renovascular and spontaneously hypertensive rats]. AB - AIM: To study the effects of praeruptorin C (pra-C), a pure constituent isolated from "Qian-Hu", the roots of Peucedanum praeruptorum Dunn. (Umbelliferae), on vascular hypertrophy, collagen content, transient [Ca2+]i, NO and vascular response of the thoracic aorta of renovascular and spontaneously hypertensive rats (RHR, SHR). METHODS: RHR and SHR were given pra-C 20 mg.kg-1.d-1 for 9 weeks, ig. Blood pressure of both rats were measured using tail cuff manometry. Under inverted microscopy the length and width of the smooth muscle cells were measured by using computer software MICC (Dongnan University). [Ca2+]i of smooth muscle cell (SMCs) was measured with Fura-2/AM. By measuring the specific aminoacid hydroxyproline content, the collagen content was obtained. By using Griess reagent, the NO in the smooth muscle cells (SMCs) was measured. RESULTS: The intermedia of the thoracic aorta in RHR was enlarged than that of the normal and pra-C groups. The size (length x width) of the SMCs of thoracic aorta from RHR increased 73.4 microns vs nomal 34.5 microns and pra-C 34 microns. The collagen content of thoracic aorta was 39% +/- 6.8% dry weight in RHR, they were 26.5% +/- 3% dry weight in normal and 25.6% +/- 1.1% dry weight in pra-C, RHR vs pra-C. The resting [Ca2+]i of single cell of SMCs was (62 +/- 6) nmol.L-1. In Hanks solution containing CaCl2 1 mmol.L-1, the resting [Ca2+]i of SMCs was (150 +/- 8) nmol.L-1 in normal. (226 +/- 11) nmol.L-1 in RHR. In presence of KCl 60 mmol.L-1, NE 10 mumol.L-1, ANG II 100 nmol.L-1 and ATP 30 mumol.L-1 the [Ca2+]i of SMCs were increased by 128%; 132%; 233% and 152% in RHR, respectively. The pra C group was similar to the normal group. The resting [Ca2+]i of SMCs was (71 +/- 6) nmol.L-1 in control of SHR, in Hanks solution containing CaCl2 1 mmol.L-1. The resting [Ca2+]i of SMCs was (160 +/- 8) nmol.L-1 in normal, and (362 +/- 18) nmol.L-1 in SHR. In presence KCl 60 mmol.L-1 and NE 10 mumol.L-1 the [Ca2+]i of SMCs were increased by 235% and 200% in SHR, respectively. Pra-C group was similar to normal group. NO of SMCs was decreased 76% in SHR, pra-C group was nearly normal. The pra-C improved vascular responses of the thoracic aorta of RHR. CONCLUSION: These results indicate that pra-C improved the vascular hypertrophy by decreasing the size of SMCs cells, collagen content. SMCs [Ca2+]i and increasing NO production. PMID- 12580081 TI - [Pharmacokinetic studies of 20(R)-ginsenoside RG3 in human volunteers]. AB - AIM: To study the pharmacokinetics of 20 (R)-ginsenoside Rg3 (GRg3) in human. METHODS: High-performance liquid chromatography-ultraviolet detection method was used in this study. RESULTS: The pharmacokinetics of GRg3 in 14 healthy volunteers were investigated. After a single oral dose of 3.2 mg.kg-1 in 8 male volunteers, the plasma concentration-time course fitted well to a two-compartment open model, with the following pharmacokinetic parameters: Tmax(0.66 +/- 0.10) h, Cmax(16 +/- 6) ng.mL-1, T1/2 alpha(0.46 +/- 0.12) h, T1/2 beta(4.9 +/- 1.1) h, T1/2(Ka)(0.28 +/- 0.04) h, AUC0-infinity(77 +/- 26) ng.mL-1.h. No kinetic analysis was made after an oral dose of 0.8 mg.kg-1 GRg3 in 6 other volunteers because of the low concentration, but a good correlation between Cmax and dosage of the two groups was found. CONCLUSION: The absorption of GRg3 was rapid in man, and its elimination was rapid after oral administration of ginsenoside Rg3. The pharmacokinetic results shows that it exhibited first order kinetic characteristics. PMID- 12580082 TI - [Characterization of cell death induced by anticancer antibiotic lidamycin in human hepatoma BEL-7402 cells]. AB - AIM: To study the features of cell death induced by the anticancer antibiotic lidamycin (LDM) in human hepatoma BEL-7402 cells. METHODS: Chromatin condensation was observed by co-staining with fluorescent dyes, hoechst 33342 and propidium iodide. "G1 sub-peak" was detected by flow cytometry and DNA ladder was observed using agarose gel electrophoresis. The caspase-3, 6 activities were measured with kits specific for them. RESULTS: Typical apoptotic chromatin condensations appeared when the BEL-7402 cells were treated with the conventional antitumor agent mitomycin C30 mumol.L-1 for 12 h. However, an abnormal type of chromatin condensation occurred when the cells were treated with LDM 1 mumol.L-1 for 6 h, which was characterized with keeping the completeness of nuclear membrane and not forming apoptotic bodies. The DNA ladder patterns were observed using agarose gel electrophoresis. The "G1 sub-peak" occurred only in the cells treated with LDM for 24 h, though chromatin condensation was earlier detected in treatment with LDM for 6 h. The caspase-3, 6 activities were increased about 5 and 4 folds, after the cells were treated with LDM 1 mumol.L-1 for 6 h, as did mitomycin C. The time of initiating chromatin condensation was earlier than that of the high peak activities of caspase-6. CONCLUSION: The characterization of cell death induced by lidamycin in the human hepatoma BEL-7402 cells differs from typical apoptosis. The results make it helpful to explain the molecular mechanism of the highly potent cytotoxicities of lidamycin toward tumor cells. PMID- 12580083 TI - [Synthesis of triphenylethylene with aliphatic cyclic moiety and its antagonism on estrogen receptor]. AB - AIM: In order to improve the biological activity and reduce the side effects and toxicity, a series of novel estrogen receptor antagonists were designed. METHODS: The key triphenylethylene intermediates were obtained by the McMurry reaction. The target compounds were prepared by etherification. The binding affinities of the target compounds for the estrogen receptor in rat uterine cytosol were measured by a competitive binding assay and their estrogen agonistic/antagonistic properties were investigated in the 3-day uterine weight assay in the immature rats. RESULTS: Thirty-five new compounds have been synthesized and their geometric configuration were determined by X-ray crystallography and 1HNMR spectral data. CONCLUSION: All of the test compounds showed affinity for the estrogen receptor (IC50 < 10(-6) mol.L-1), especially compound 35 with IC50 1.07 x 10(-8) mol.L-1. Some compounds are antagonists, inhibiting uterus growth; others are agonists, promoting uterus growth. Compounds 14 and 27 are superior antagonists to tamoxifen. PMID- 12580084 TI - [Synthesis and anti-tumor activities of 1,4-bis[3-(amino-dithiocarboxy)propionyl] piperazine derivatives]. AB - AIM: To synthesize piperazine derivatives and screen anti-tumor compounds with higher activity and lower toxicity. METHODS: Selecting 1,4-bis(3 bromopropionyl)piperazine as leading compound, a series of 1,4-bis[3-(amino dithiocarboxy)propionyl] piperazine derivatives (4a-j) were synthesized through the use of aminodithiocarboxylate. All the synthetic compounds (4a-j) were tested for their anti-tumor activity against eight kinds of tumor cells. RESULTS: Compounds (4a-j) are new compounds, among them, compounds 4c, 4d and 4e showed anti-tumor activity against HL-60. The inhibition of compounds 4c, 4d and 4e against HL-60 are 44%, 90% and 70% respectively, at the concentration of 10 mumol.L-1. However, the inhibition of the other kinds of anti-tumor cells are not distinctive. CONCLUSION: These results suggest that this may be one of the effective routes to improve the anti-tumor activity and reduce the toxicity of 1,4-bis(3-bromopropionyl)piperazine. PMID- 12580085 TI - [Synthesis, preliminary antigastrelcosis activity and the protective effect on acute liver injury of zinc chlorin E4]. AB - AIM: To study the synthesis of zinc chlorin e4(1), its experimental antigastrelcosis activity as well as the protection against acute liver injuries. METHODS: Chlorin e6(3) was prepared through acidic and alkaline oxidative degradation using silkworm excrement crude chlorophyll extracts as starting material. Compound 1 was synthesized via Zn(OAc)2 complex action with Chlorin e4(2) which was prepared by refluxing 3 in pyridine. Gastric ulcers were induced by abdominal injection of 0.2% indomethacin at 20 mg.kg-1 in rats. The ulcer indexes and ulcer numbers in gastric mucosa were determined. Acute liver injuries were induced by abdominal injection of 0.3% thioacetamide (TAA) or 0.3% CCl4 at 20 mg.kg-1 in mice, and activities of SGPT in mice were determined. RESULTS: Compound 1 is previously unknown. Compared with control group, abdominal administration of 1 at 100 mg.kg-1 reduced significantly the gastric ulcer index (P < 0.001) and the number of ulcer (P < 0.001) induced by indomethacin in rats. Abdominal administration of 1 at 100 mg.kg-1 x 3 exhibited marked inhibitory effects on elevated activities of SGPT induced by TAA (P < 0.02) or CCl4(P < 0.01) in mice. CONCLUSION: These results show that 1 has significant protective effect against indomethacin-induced gastric lesion in rats and TAA or CCl4 induced acute liver injuries in mice. It is suggested that 1 may be a promising new drug candidate for antigastrelcosis and liver injury protection. PMID- 12580086 TI - [Studies on the polysaccharide H3 of Cuscuta chinensis Lam]. AB - AIM: To study the structural characterization of an acidic polysaccharide, H3, isolated from the seeds of Cuscuta chinensis Lam. METHODS: Both chemical (such as sugar component analysis, methylation analysis, reduction of uronic acid, partial acidic hydrolysis, etc.) and spectral analysis (1HNMR, 13CNMR, IR, etc.) were used to investigate the structural characterization of H3. RESULTS: The molecular weight of H3 was estimated to be more than 1.0 x 10(6). The analytical results of sugar components indicated that H3 composed of Rha, Ara, Gal, and Gal A. Methylation analysis, partial acidic hydrolysis and 1H, 13CNMR further identified the linkages and sequence of the residues of H3. CONCLUSION: These results suggest that H3 is a highly branched heteropolysaccharide with structure elucidation for the first time. PMID- 12580087 TI - [Studies on the active polysaccharides from Lycium barbarum L]. AB - AIM: To investigate the structures and immunomodulation activity of four homogeneous polysaccharides: LBP 1a-1, LBP 1a-2, LBP 3a-1 and LBP 3a-2 isolated from Lycium barbarum L. brought from Zhongning County, Ningxia Province. METHODS: Their molecular weights, sugar component (constituents) and their linkages were determined by gel permeation chromatography, acid hydrolysis, periodate oxidation and NMR spectrum. The activity of immunomodulation was evaluated with splenocyte proliferation by [3H]-TDR incorperation, in vitro. RESULTS: Four polysaccharides with molecular weights 11.5 x 10(4), 9.4 x 10(4), 10.3 x 10(4) and 8.2 x 10(4), were shown to enhance splenocyte proliferation induced by ConA. LBP 1a-1 and LBP 1a-2 were alpha-(1-->6)-D-glucans. LBP 3a-1 and LBP 3a-2 were found to be a-(1- >4)-D-polygalacturonans. CONCLUSION: The four polysaccharides were first isolated from this plant. Polysaccharides with main chain of alpha-(1-->4)-D polygalacturonans showed stronger immunomodulation activity. PMID- 12580088 TI - [Cochinchinenin--a new chalcone dimer from the Chinese dragon blood]. AB - AIM: To study the active constituents of Dracaena cochinchinensis (Lour.) S.C. Chen. in the commercial dragon blood. METHODS: Various column chromatographies with Sephadex L-20 gel, MCI gel and silica gel were employed for the isolation and purification. The structures of compounds were elucidated by spectral analysis. RESULTS: Nine chalcones were isolated from the commercial dragon's blood which was made of D. cochinchinensis (Lour.) S.C. Chen.. By means of spectral data, they were identified as 1-[5-(2,4,4'-trihydroxydihydrochalconyl)] 1- (p-hydroxyphenyl)-3-(2-methoxy-4-hydroxy-phenyl)-propane (1), 2' methoxysocotrin-5'-ol (2), socotrin-4'-ol (3), 2-methoxy-4, 4' dihydroxydihydrochalcone (4), 2, 4, 4'-trihydroxy-dihydrochalcone (5), 2, 4, 4' trihydroxy-6-methoxydihydrochalcone (6), 2', 4', 4-trihydroxychalcone (7), 2 methoxy-4, 4'-dihydroxychalcone (8) and 2'-methoxy-4', 4-dihydroxychalcone (9). CONCLUSION: Compound 1 is a new chalcone dimer and named as cochinchinenin. Compounds 2-9 were isolated from D. cochinchinensis (Lour.) S.C. Chen. for the first time. PMID- 12580089 TI - [Study on the phase I metabolites of phenoprolamine hydrochloride in rat bile by LC/DAD/MSD]. AB - AIM: To study the phase I metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. METHODS: DDPH was administered i.p. to bile duct-cannulated rats. Bile samples were collected before administration and up to 12 h after administration. After being treated with beta-glucuronidase, the bile samples were purified and enriched with C-18 SPE columns, and then were analyzed by LC/DAD/MSD. The samples containing synthesized reference standards of DDPH metabolite 1-(2, 6 dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethylamino)-propane (M1), 1-(2, 6 dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxy-phenylethylamino)-propane (M2), 1-(2,6 dimethyl-4-hydroxyphenoxy)-2-(3,4- methoxyphenylethylamino)-propane (M3), 1-(2, 6 dimethyl-3-hydroxyphenoxy)-2-(3-hydroxy-4- methoxyphenylethylamino)-propane (M4), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2- (3-hydroxy-4-methoxyphenylethylamino) propane (M5) and 1-(2,6-dimethyl-4-hydroxyphenoxy)-2-(3-methoxy-4- hydroxyphenylethylamino)-propane (M6) were analyzed by LC/DAD/MSD under identical conditions. RESULTS: The retention times, UV spectra, molecular weights and production spectra (obtained by collision-induced dissociation) of the apparent ions of peak A, B, C, D, E and F in the total ion chromatogram of DDPH treated rat bile sample were consistent with those of M1, M2, M3, M5, M4 and M6, respectively. CONCLUSION: M1, M2, M3, M4, M5 and M6 were identified as the phase I metabolites of DDPH in the rat. PMID- 12580090 TI - [Study on photodegradation kinetics of lomefloxacin hydrochloride aqueous solution]. AB - AIM: To investigate the photodegradation kinetic characteristics of lomefloxacin hydrochloride aqueous solution. METHODS: The HPLC method was used to determine the photodegradation kinetic parameters of lomefloxacin hydrochloride aqueous solution (10 micrograms.mL-1) under various pH buffer solutions, various ionic strength of different ions and various dielectric constant conditions. RESULTS: With comparative regression analysis of linear fit, the photodegradation kinetic order of lomefloxacin hydrochloride aqueous solution was determined as n = 1. The lomefloxacin hydrochloride aqueous solution is most unstable at pH 5.08-9.40 and is relatively stable at pH 2.02-5.08 and 9.40-11.10. The higher the ionic strength in lomefloxacin hydrochloride aqueous solution, the higher the photodegradation kinetic rate constant is. The chloride ion has important effect on photodegradation in aqueous solution of lomefloxacin and results in producing chlorine derivative as the highest photodegradation product. As the dielectric constant of solution increased, the photodegradation kinetic rate constant was also increased. CONCLUSION: It was found that the photodegradation of lomefloxacin followed apparent first-order kinetics. The photodegradation kinetic rate was affected by pH remarkably and showed positive correlation with ionic strength and dielectric constant. PMID- 12580091 TI - [Analysis of the essential oil of Schisandra chinensis (Turcz.) Baill. with GC/MS]. AB - AIM: To detect chemical components of the essential oil of Schisandra chinensis (Turcz.) Baill.. METHODS: The essential oil was analyzed with GC/MS, heuristic evolving latent projections(HELP) resolution and overall volume integration method. RESULTS: HELP method along with the data from GC/MS can be used to conduct the peak purity examination and resolution of overlapping peaks to obtain pure chromatogram and MS spectrum of each component. Fifty six components were separated and 49 of them were qualitatively and quantitatively analyzed. All the components represent about 98.27% of the total content. CONCLUSION: The resolved pure chromatogram and MS spectrum can greatly enhance the reliability of similar searches in the commercial MS database and thus contribute to the accuracy of the qualitative and quantitative analysis of the essential oil. PMID- 12580092 TI - [Tetrandrine loaded sustained-release microcapsules for lung targeting]. AB - AIM: To prepare lung targeted tetrandrine (TET) loaded sustained-release drug delivery system by microencapsulation, decrease the toxicity and enhance the therapeutic function of anti-pulmonary hypertension of TET. METHODS: Albumin microcapsules were produced by spray drying-thermal denaturation, a new technique. Some characterization of the prepared microcapsules was evaluated. Distribution of the microcapsules and their anti-pulmonary hypertension effect in vivo were investigated. RESULTS: The spherical microcapsules showed a drug loading of 37.88%. Compared to the original drug, the rate of TET released from the positively charged microcapsules in vitro was significantly decreased and fitted well by Higuchi equation. The TET concentrations in mouse lungs of TET microcapsules were significantly higher than those of TET injection, and the mean retained time of TET in lungs was prolonged from 157.1 h to 223.6 h after microencapsulation. The in vitro--in vivo correlation was established and confirmed (P < 0.001). CONCLUSION: The new spray drying-thermal denaturation method allows the preparation of drug loaded albumin microcapsules with desired results. The prepared microcapsules were found to have the potential function of delivering TET to pulmonary artery via i.v., with low toxicity and high efficacy. PMID- 12580093 TI - [Biotransformation of (+)- and (-)-clausenamide in rats]. AB - AIM: To study the metabolic pathway of chiral clausenamide in the rat and understand its stereoselectivity. METHODS: The urine, feces and blood of rat were gathered after the drug was administered, the known metabolites were analyzed by HPLC-DAD and one unknown metabolite was elucidated by using LC-MS analysis. Metabolic stereoselectivity was determined by comparing the metabolic results of (+)- and (-)-clausenamide. RESULTS: Six known metabolites were determined and one unknown metabolite was elucidated as N-demethylclausenamide. The metabolic stereoselectivity was shown distinctly. CONCLUSION: Chiral clausenamide was mainly metabolized by hydroxylation in liver and the biotransformation exhibited pronounced substrate stereoselectivity. PMID- 12580094 TI - [Identification of Chinese crude drug snake gallbladder by DNA molecular marker]. AB - AIM: It is difficult to identify the Chinese crude drug snake gallbladder accurately by morphological and microscopical characteristics or chemical components only. In order to solve the problem, the technique based on DNA molecular marker was introduced into the authentication of snake gallbladder. METHODS: DNA templates were extracted from the membrane or the bile of snake gallbladder, and also from the muscle of the original animal Elaphe schrenckii. About 400 bp DNA fragments of 12S rRNA gene were amplified from the templates and sequenced subsequently. RESULTS: Enough amounts of DNA templates could be extracted from a bit of membrane or bile of snake gallbladder. The sequence of amplicons from the membrane, bile and muscle of the same individual were identical completely. CONCLUSION: The technique of DNA molecular marker could be used for the authentication of snake gallbladder and bile. The results indicate that the technique could be used for the identification of crude drugs from other animal secretion. DNA sequence analysis also demonstrated that the origins of commercial snake gallbladder were complicated and more efficient quality control was necessary for supervising the crude drug in the market. PMID- 12580095 TI - [Study on the essential oil components in Thymus quinquecostatus Celak]. AB - AIM: To analyse the essential oil components of Thymus quinquecostatus Celak. METHODS: Gas chromatography/mass spectrometry were used. RESULTS: A total of 78 components were separated. 34 of them were identified, which accounted for 95.93% of the total peak area. The main constituents are o-cymene, carvacrol, caryophyllene, 2-isopropyl-1-methoxy-4-methylbenzene and gamma-terpene. CONCLUSION: Thymus quinquecostatus Celak has perfume and medical development value. PMID- 12580096 TI - [Progress in oral osmotic pump tablets for poorly water soluble drugs]. PMID- 12580097 TI - [Influences of melatonin on the growth of HELA cells]. AB - AIM: To investigate the influence of melatonin (MT) on growth of HeLa cells in vitro. METHODS: The antiproliferation activities of MT were evaluated on HeLa cells by means of trypan blue dye exclusion and MTT vital staining. The morphological changes of MT on HeLa cells were observed under transmission electron microscope. Cell division cycle influenced by MT was assessed by a flow cytometer. RESULTS: MT produced a certain inhibition on HeLa cells at the concentration of 2 mmol.L-1 and prolonged the time of double amount (TD). The fraction of cells inhibited was 61.0%. The IC50 of HeLa cells exposed to MT for 96 h was 2.039 mmol.L-1. The flow cytometric analyses showed that exposure to MT for 72 h reduced the number of HeLa cells in phase S. Under electronic microscope, the HeLa cells exposed to MT for 72 h displayed morphological changes of necrosis, apoptosis, more heterochromosome and less somatic chromosome. CONCLUSION: MT showed certain influence on HeLa cells. Its mechanism may probably be attributable to reduction of the number of cells in phase S. PMID- 12580098 TI - [Transportation of the enantiomers of trans tramadol and O-demethyltramadol across blood-brain barrier]. AB - AIM: To investigate the transportation of the enantiomers of trans tramadol (trans T) and its active metabolite, O-demethyltramadol (M1) across blood-brain barrier. METHODS: Rats were sacrificed by femoral artery bleeding 1 h after i.p. administration of trans T hydrochloride, 16.7 mg.kg-1 or 50.0 mg.kg-1. Blood, cerebrospinal fluid and cerebral cortex were taken out. The enantiomers of trans T and M1 were analyzed by high performance capillary electrophoresis (HPCE). RESULTS: Among the three tissues, the concentration of each enantiomer of trans T and M1 was the highest in the cerebral cortex, and the lowest in the cerebrospinal fluid. In the serum, the concentration of (+)-trans T was higher than that of (-)-trans T, and the concentrations of the enantiomers of M1 were similar. In the cerebrospinal fluid and cerebral cortex, the concentration of (+) trans T was higher than that of (-)-trans T, and the concentrations of (+)-M1 was lower than that of (-)-M1. CONCLUSION: The transportation across blood-brain barrier of the enantiomers of trans T and M1 was stereoselective. In the brain tissues, the concentrations of (+)-trans T and (-)-M1 were higher than those of their enantiomers. PMID- 12580099 TI - [Antifibrotic effects of genistein and quercetin in vitro]. AB - AIM: To study the antifibrotic effects of genistein (GE) and quercetin (QU) on rat hepatic stellate HSC-T6 cell proliferation stimulated with platelet-derived growth factor (PDGF), collagen synthesis and type I procollagen messenger RNA (mRNA) expression stimulated with transforming growth factor beta 1 (TGF beta 1). METHODS: Cell proliferation was measured by crystal violet staining assay. Collagen synthesis was determined by 3H-proline incorporation assay. Type I procollagen mRNA level was determined by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: GE (25-70 mumol.L-1) and QU (6.25-50 mumol.L-1) concentration-dependently attenuated PDGF-drive HSC-T6 cell proliferative activity. TGF beta 1-stimulated collagen synthesis was also reduced. This was associated with a decrease of type I procollagen mRNA, indicating an effect at a pretranslational level. CONCLUSION: GE and QU may have therapeutic potential against liver fibrosis by regulating PDGF and TGF beta 1 actions. PMID- 12580100 TI - [Effects of opioids on Ca2+/calmodulin dependent protein kinase signal pathway in NG108-15 cells]. AB - AIM: To observe the change of Ca2+/calmodulin dependent protein kinase II (CaMK II) signal pathway in opioid dependent NG108-15 cells. METHODS: NG108-15 cells were used as an in vitro model system. Competitive protein binding assay and radioimmunoassay were used to examine the intracellular cAMP accumulation. Calmodulin activity was assayed by PDE method. CaMK II activity was assayed by gamma-32 P incorporation of syntide-2. RESULTS: DPDPE long-term treatment increased calmodulin activity and CaMK II activity in both cytoplasm and nucleus of NG108-15 cells. Specific calmodulin antagonist W-7 was found to significantly inhibit the elevation of calmodulin and CaMK II activity which resulted from DPDPE long-term treatment, and CaMK II inhibitor KN-62 also inhibited elevation of CaMK II activity by DPDPE long-term treatment. When naloxone was added to NG108-15 cells which were long-term treated by DPDPE, calmodulin and CaMK II activity increased, indicating that naloxone withdrawal can increase Ca2+/CaMK II pathway activity. CONCLUSION: The results indicate that Ca2+/CaMK II pathway was involved in the mechanisms of opioids dependence when DPDPE was long-term administered to NG108-15 cells. PMID- 12580101 TI - [Studies on the synthesis and bioactivity of alpha-alkylaminobenzyl-phosphonic acids]. AB - AIM: To search for some substituted benzyl phosphonic acids as leading compounds with inhibiting effect on osteoclast formation. METHODS: Target compounds were prepared from aromatic aldehydes, primary amine and phosphorous acid using tetramethylenesulfone as solvent via Arbuzov type reaction. The effect on inhibiting the formation of osteoclast-like cells (OLC) of related compounds was studied by incubating the extract of rat femur marrow. RESULTS: Ten compounds of alpha-alkylaminobenzyl phosphonic acids have been synthesized and identified by MS or 1HNMR analysis. Three (2, 8 and 9) of them were found to have notable effect on the inhibition of OLC formation (P < 0.01). CONCLUSION: Among the present substituted benzyl phosphonic acids, the increased aromaticity and hydrophobicity (such as compound 9) can remarkably enhance the ability to inhibit OLC formation. PMID- 12580102 TI - [Triterpenoid saponins from Eclipta prostrata L]. AB - AIM: To study the triterpenoid saponins in the Chinese traditional medicine Eclipta prostrata L.. METHODS: Column chromatography with silica gel and HPLC were employed for the isolation and purification. The molecular structures were determined on the basis of spectral analysis (IR, MS, 1HNMR, 13CNMR, HMQC and HMBC). RESULTS: Two new triterpenoid saponins, named eclalbasaponins XI (4) and XII (5), were obtained and their structures were elucidated as 3-O-[beta-D glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-16 alpha-ethoxy-olean-12-ene-28-oic acid-28-O-beta-D-glucopyranoside and 3-O-[(2-O-sulfuryl-beta-D-glucopyranosyl) (1 ->2)-beta-D-glucopyranosyl]-echinocystic acid-28-O-beta-D-glucopyranoside, respectively, along with three known saponins, eclalbasaponins II (1), I (2) and III (3). CONCLUSION: Compounds 4 and 5 are new compounds, 1 and 5 induced morphological deformation of Pyricularia oryzae mycelia. PMID- 12580103 TI - [Studies on chemical structure of polysaccharide from fruit body of Coriolus versicolor]. AB - AIM: To study the structure of polysaccharides from Coriolus versicolor. METHODS: By chromatography a homogeneous polysaccharide B-1-3 was obtained. The structure of B-1-3 was elucidated using the method of total acid hydrolysis, methylation, and 1D and 2D NMR. RESULTS: B-1-3 was found to be mainly composed of Glc by total acid hydrolysis. Its molecular weight was estimated to be 3.16 x 10(5) by HPLC method. Methylation analysis and 1H and 13CNMR indicated the main chain of B-1-3 consisted of beta-D-1,4-Glc and beta-D-1,3-Glc, and branch chains were situated, beta-D-1,3,6-Glc and beta-D-1,4,6-Glc. 1H and 13CNMR also showed that beta-D-Glc 1-->3-beta-D-Glc-beta-D-Glc-1-->4-beta-D-Glc and beta-D-Glc-1-->4-beta-D-Glc-beta D-Glc-1-->4-beta-D-Glc fragments existed in B-1-3. CONCLUSION: B-1-3 is a glucan which main chain was composed of beta-D-1,4-Glc and beta-D-1,3-Glc beta-D-1,6 Glc. PMID- 12580104 TI - New biflavanones and bioactive compounds from Stellera chamaejasme L. AB - AIM: To study the chemical constituents of the root of Stellera chamaejasme L. METHODS: Various column chromatographies on silica gel and RP-18 were employed for isolation and purification. Structures of compounds were elucidated by spectral analysis. RESULTS: Eight lignans and three biflavonoids possessing a C 3/C-3" linkage were isolated. They are ruixianglangdusu A (1) and B (2), 4',4'",5,5",7,7"-hexahydroxy-3,3"-biflavone (3), (+)-kusunokinin (4), lirioresinol-B (5), magnolenin C (6), (-)-pinoresinol monomethyl ether (7), (-) pinoresinol (8), (+)-matairesinol (9), isohinokinin (10) and (-)-eudesmin (11). CONCLUSION: Compounds 1 and 2 are new biflavanones, 1 is enantiomeric to known chamaejasmenin C, 4, 6, 8, 9, 10 and 11 were isolated from this plant for the first time, and 7 was isolated from natural resources for the first time. In vitro bioassays showed that 3 and 8 exhibited antibacterial activity, and 1, 2, 9 and 11 exhibited immunomodulatory activity. PMID- 12580105 TI - [Selective isolation of anethole from volatile oil of Foeniculum vulgare Mill by inclusion crystalline with chela-shape host]. AB - AIM: To isolate the components from the volatile oil of Foeniculum vulgare Mill. METHODS: According to the function of molecular recognition of supramolecular chemistry, chela shape molecule, trans-1, 2-biphenyl-1, 2-acenaphthendiol was used as host molecule and the volatile oil of Foeniculum vulgare Mill as guest molecule. Trans-1, 2-biphenyl-1, 2-acenaphthendiol can recognize the components that endowed with interactional complementarity and form inclusion compound as crystals. RESULTS: The anethole in the volatile oil was selectively included as trans-1,2-biphenyl-1,2-acenaphthendiol which was obtained in pure state from the inclusion compound by Kugelrohr vacuum technology. The formation of inclusion compound was confirmed by means of IR and powder XRD. The structure of the selectively isolated component was elucidated as trans-anethole by means of IR, 1HMMR and MS. CONCLUSION: The experimental results showed that the method is simple, rapid and selective for isolation anethole from volatile oil of Foeniculum vulgare Mill. PMID- 12580106 TI - [Application of FAM networks in recommendation of mobile phase for RP-HPLC]. AB - AIM: To establish expert system to recommend solvent strength in RP-HPLC by using FAM (fuzzy associate memorizer) networks. METHODS: Symbolic rules reflecting the quantitative relationship among molecular structure, capacity factors and solvent strength in RP-HPLC were extracted from trained radial basis function networks. Then FAM networks consisting of these rules were built to recommend mobile phase strength for RP-HPLC. RESULTS: The validation results of the system by monocomponent and mutilcomponent drug samples were satisfied. CONCLUSION: The mobile phase recommendation system for RP-HPLC analysis of drugs showed reliable performance. PMID- 12580107 TI - [Flow injection biamperometric analysis of isoniazid]. AB - AIM: To establish a simple, rapid, and accurate electrochemical method for on line determination of isoniazid. METHODS: Based on the flow injection biamperometry for irreversible couple system, and using two preanodized platinum electrodes with the applied potential difference of 0 V, the biamperometric method for the determination of isoniazid has been proposed by coupling the catalytic oxidation of isoniazid and the reduction of platinum oxide. RESULTS: Common excipients, inorganic ions, amino acids, vitamins and proteins do not interfere with the determination. Linear relationship between current and the concentration of isoniazid is obtained in the range of 1.0 x 10(-6)-1.0 x 10(-4) mol.L-1 (gamma = 0.998, n = 11). The RSD of 1.8% was obtained for 8 successive determinations of 1.0 x 10(-5) mol.L-1 isoniazid. CONCLUSION: The proposed method has been shown to be sensitive, selective, rapid (120 samples.h-1), and suitable for the on-line direct determination of isoniazid. PMID- 12580109 TI - [Rapid analysis of terbutaline by combined solid phase extraction/liquid chromatography tandem mass spectrometry]. AB - AIM: To develop a liquid chromatography-electrospray ionization tandem mass spectrometry method for rapid analysis of terbutaline at level of 50 pg.mL-1 in human plasma. METHODS: Samples containing terbutaline and salbutamol (internal standard, IS) were extracted using C18 solid-phase extraction cartridges, followed by liquid chromatographic separation and mass spectrometric detection. The mobile phase consisted of acetonitrile-water-formic acid (20:80:1), at a flow rate of 0.4 mL.min-1. Selected reaction monitoring with mass transitions m/z 226- >151 and m/z 240-->148 were used for terbutaline and IS, respectively. RESULTS: The chromatographic analysis time for each sample was approximately 3.8 min. The assay was linear from 0.05 to 8.0 ng.mL-1. The between-run precision and accuracy of the quality controls (QCs, 0.1, 0.4 and 4.0 ng.mL-1) were characterized by relative standard deviation (RSD) of 2.5% to 7.1% and relative errors of -3.1% to 5.7%, respectively. The within-run precision of QCs was characterized by RSD of 4.2% to 6.6%. This method was applied to the analysis of samples taken up to 60 h after oral administration of 10 mg bambuterol in healthy volunteers. CONCLUSION: The method is shown to be suitable for clinical investigation of terbutaline pharmacokinetics, which offers advantages of specificity, speed, and greater sensitivity over previously reported methods. PMID- 12580108 TI - [Determination of annonaceous acetogenins in annonaceae plants by HPLC]. AB - AIM: To develop a method for analysis of antitumor annonaceous acetogenins in Annonaceae plants by HPLC. METHODS: Squamostatin-B (1), squamocin (2) and annonin VI (3) were used as standard substances. Chromatography column was a Rp-18; the mobile phase was methanol-water (90:10); the flow rate was 1.0 mL.min-1 and the detecting wavelength was 220 nm. RESULTS: A linear range was obtained from 2.3 to 13.8 micrograms with a good correlation. The recoveries of (1), (2) and (3) were 100.3%, 100.3% and 100.0%, respectively. CONCLUSION: This method was developed for the analysis of acetogenins by HPLC for the first time. The method is rapid, accurate and suitable for the analysis of the antitumor acetogenins in Annonaceae plants. PMID- 12580110 TI - [Application of an artificial neural network in the design of sustained-release dosage forms]. AB - AIM: To use the artificial neural network (ANN) in Matlab 5.1 tool-boxes to predict the formulations of sustained-release tablets. METHODS: The solubilities of nine drugs and various ratios of HPMC: Dextrin for 63 tablet formulations were used as the ANN model input, and in vitro accumulation released at 6 sampling times were used as output. RESULTS: The ANN model was constructed by selecting the optimal number of iterations (25) and model structure in which there are one hidden layer and five hidden layer nodes. The optimized ANN model was used for prediction of formulation based on desired target in vitro dissolution-time profiles. ANN predicted profiles based on ANN predicted formulations were closely similar to the target profiles. CONCLUSION: The ANN could be used for predicting the dissolution profiles of sustained release dosage form and for the design of optimal formulation. PMID- 12580111 TI - [Long-term immunogenicity and immunologic priming of animals to pulsed controlled release system for tetanus toxoid vaccine delivery]. AB - AIM: To evaluate the immunogenicity of tetanus toxoid (TT) encapsulated in biodegradable polymer microspheres composed of polylactide (PLA). METHODS: The antibody levels elicited by microsphere formulations in mice for 1 year were examined, the anamnestic responses to a low dose booster 1 year after priming were also examined. RESULTS: Microsphere formulations made from PLA were characterized by pulse controlled-release models, differing in polymer molecular weight, protein loading and particle size of the microspheres. Microsphere formulations elicited significantly higher IgG antibody levels than a single injection of soluble TT. The antibody levels elicited by microsphere formulations were similar to those elicited by three doses of soluble TT. CONCLUSION: A single dose tetanus toxoid based on pulsed release from biodegradable and biocompatible polymer microspheres has been developed. The formulations showed great benefits and pharmaceutical application. PMID- 12580115 TI - [Progress in studies on antidiabetic agents]. PMID- 12580112 TI - [Relative bioavailability of effervescent tablet of potassium chloride in healthy volunteers]. AB - AIM: To probe the approach by which the pharmacokinetics and relative bioavailability of endogenous medicinal substances can be studied. METHODS: A randomized three-crossover study was performed in 18 healthy male volunteers. In two of the three study periods, a single 2 g dose of either effervescent tablet or common tablet of potassium chloride was administered; whereas in one of three periods no drug treatment was given to allow the nondrug-related (endogenous) potassium in urine to be determined. In each period the urine samples were collected at the following intervals: 0-2, 2-4, 4-6, 6-8, 8-10, 10-12, 12-24, 24 48 h after dose. Urine potassium was determined and the cumulative urine potassium-time data were fitted to a one-compartment model with first-order absorption. Bioavailability was represented by cumulative amount of potassium excreted in urine during 48 hours after drug administration and the bioequivalence of the two formulations was evaluated by analysis of variance and two one-sided t-test. RESULTS: The pharmacokinetic parameters were as follows: effervescent tablet T1/2 ke = (6 +/- 5) h, T1/2 ka = (0.08 +/- 0.08) h, ku = (0.09 +/- 0.04) h-1, Xmax/f = (18 +/- 8) mmol; common tablet T1/2 ke = (8 +/- 5) h, T1/2 ka = (0.11 +/- 0.11) h, ku = (0.07 +/- 0.04) h-1, Xmax/f = (18 +/- 8) mmol. Relative bioavailability of effervescent tablet was 97.5% +/- 15.2% compared with common tablet. CONCLUSION: The two formulations were of bioequivalence. The methods used in this study might be applicable to other similar studies involving endogenous medicinal substances. PMID- 12580114 TI - [Effect of pH on the permeability of timolol maleate across isolated rabbit cornea]. AB - AIM: To study the effect of pH on the transcorneal permeability of timolol maleate (TM). METHODS: The apparent distribution coefficients of TM were determined. The permeability of TM across isolated rabbit cornea was measured using in vitro method at various pH. RESULTS: The partition coefficient and pKa of TM were 63.63 and 9.17 respectively. At neutral pH, the apparent permeation coefficient of TM was 1.43 x 10(-5) cm.s-1. When the pH varied from 6.65 to 9.20, the cumulative amount for timolol transcorneal penetration increased 1.3 times and the lag time decreased more than 19-folds. The calculated permeability coefficients of ionized and un-ionized timolol were 1.29 x 10(-5) cm.s-1 and 4.22 x 10(-5) cm.s-1, respectively. CONCLUSION: Timolol penetrated corneal membrane mainly as free base by intracellular pathway, and corneal epithelium was the rate limiting barrier. PMID- 12580113 TI - [Population pharmacokinetic model for gentamycin and its predictive value]. AB - AIM: To construct a population pharmacokinetic model to describe gentamycin concentrations in serum in newborn infants and to validate the predictive ability of this model. METHODS: Data used in this study were obtained from 30 neonates with 80 serum samples. A one-compartment open model was used to describe the kinetics of gentamycin after intravenous infusion. Following Sheiner's idea of population pharmacokinetics, a programs to estimate population parameter and individual parameter of gentamycin was made. The target function minimality was obtained from Monte Carlo algorithm. The predictive ability of the developed model was evaluated by computing precision and accuracy of serum concentration predicted using the parameter estimates. RESULTS: Fitted population pharmacokinetic parameters (mean +/- standard deviation) were as follows: Ke: 0.220 +/- 0.022 h-1, Vd: 0.51 +/- 0.06 L.kg-1, Cl: 112 +/- 10 mL.h-1.kg-1. For the population analysis sample and the predictive sample, predicted and observed concentrations were all close with correlation coefficient 0.920 and 0.946, respectively. Mean prediction error (ME) and root mean squared error (RMSE) were 0.001 mg.L-1 and 0.84 mg.L-1 for the predictive sample, respectively. CONCLUSION: Observed gentamycin serum concentrations were explained very well by this model. We propose the use of this population pharmacokinetic model to optimize gentamycin clinical therapies in our institution and others with similar patient population characteristics. PMID- 12580116 TI - [Advances in antitumor activity of the hsp90 inhibitor geldanamycin]. PMID- 12580117 TI - Ideology and trust: a reply to Bloche. PMID- 12580119 TI - [Current status of the treatment of chronic heart failure--based on EBM]. PMID- 12580120 TI - [Current status and management of chronic kidney failure]. PMID- 12580118 TI - [The current status and future prospect of vaccines--with special reference to respiratory tract infections]. PMID- 12580122 TI - [Clinical application of hematopoietic factors]. PMID- 12580121 TI - [Fluid balance of patients undergoing dialysis]. PMID- 12580124 TI - [Physiopathology and treatment of alcoholic liver dysfunctions]. PMID- 12580123 TI - [Respiratory tract diseases and pulmonary surfactant protein molecules]. PMID- 12580125 TI - [Physiopathology and molecular mechanism involving systemic lupus erythematosus]. PMID- 12580126 TI - [Septicemia, the current concept]. PMID- 12580127 TI - [Steroid therapy and osteonecrosis]. PMID- 12580128 TI - [Visualization of human brain activities--progress in the non-invasive methods to detect cerebral functions]. PMID- 12580129 TI - [Abnormal bone calcium metabolism]. PMID- 12580130 TI - [Forefront of the treatment of severe, acute pancreatitis]. PMID- 12580131 TI - [Standard treatment of malignant lymphoma]. PMID- 12580132 TI - [Clinical description and guidelines concerning COPD]. PMID- 12580133 TI - [Ambulatory blood pressure monitoring (ABPM) and management of hypertension]. PMID- 12580134 TI - [Physiopathology and drug therapy of type 2 diabetes mellitus]. PMID- 12580135 TI - Liaison lecturers have a fundamental role. PMID- 12580136 TI - Nursing rivalry is not good practice. PMID- 12580137 TI - Problems regarding skin care of children. PMID- 12580144 TI - Maurice Hofnung. From bacterial genetics to immunology: the route of an enlightened scientist. Proceedings of a meeting. March 10-12, 2002, Paris, France. PMID- 12580145 TI - Proceedings of the International Meeting on Aicardi-Goutieres Syndrome. Pavia, Italy, 28-29 May 2001. PMID- 12580147 TI - Tracking health care costs: hospital spending spurs double-digit increase in 2001. AB - This Data Bulletin is based on data from the Milliman USA Health Cost Index ($0 deductible), which is designed to reflect claims increases faced by private insurers; the Kaiser Family Foundation/Health Research and Educational Trust survey of employer-based health plans for 1999- 2002; the KPMG survey of employer based plans for 1991-98; and the U.S. Bureau of Labor Statistics Employment, Hours and Earnings series to track payroll costs and Hospital Producer Price Index to track hospital prices. PMID- 12580148 TI - Aging plays limited role in health care cost trends. AB - This Data Bulletin analyzes changes in an aging index that measures the extent to which average health care spending per person under age 65 changes from year to year due solely to the aging of the U.S. population, while holding all other cost drivers constant. PMID- 12580146 TI - Some communities make progress in reducing children's uninsurance. AB - This Data Bulletin presents findings from the Household Survey, a nationally representative telephone survey of the civilian, noninstitutionalized population, as well as 12 randomly selected communities conducted as part of the Community Tracking Study. The 1996-1997 survey includes nearly 33,000 families and 60,000 individuals, and the 1998-1999 survey includes approximately 32,000 families and 59,000 individuals. PMID- 12580149 TI - Adherence to antiretroviral therapy. PMID- 12580150 TI - Third European Stanley Foundation Conference on Bipolar Disorder. 12-14 September 2002. Freiburg, Germany. Proceedings and abstracts. PMID- 12580151 TI - Non-invasive ventilation in acute exacerbations of chronic obstructive pulmonary disease: a new gold standard? PMID- 12580152 TI - Neuromuscular blockade studies of critically ill patients. PMID- 12580153 TI - Antifungal prophylaxis for intensive care unit patients: let's fine tune it. PMID- 12580154 TI - Management and outcome of pregnancies complicated with adnexal masses. AB - Our purpose was to evaluate the pathologic features and outcome of pregnancies that were complicated with adnexal masses and were managed surgically. A review of patients who had persistent adnexal masses during pregnancy and needed surgical removal of tumours was performed from January 1998 to April 2001. There were 14 cases of persistent adnexal masses identified among 2000 deliveries. There were 13 patients who had surgical interventions: nine (69.2%) had surgery during ongoing pregnancy (at mean gestational age 17+/-3.7 weeks), two (15.3%) with caesarean section, one (7.6%) after evacuation of missed abortion and one (7.5%) after delivery. Out of 13, ten (76.9%) were benign [mature cystic teratoma, six (46.9%); serous cyst adenoma, two (15.3%); mucinous cyst adenoma, one (7.6%); paratubal cyst, one (7.6%)] and three (23%) were malignant (one immature teratoma, one papillary cyst adenocarcinoma and one krukenberg tumour]. Both patients operated on after 24 weeks had pre-term delivery. The worst outcome in the form of PPROM and preterm delivery at 28 weeks occurred in a patient who underwent emergency surgery. The incidence of malignancy was four- to fivefold greater in our series than reported in the literature. Ultrasound was unable to distinguish malignant cases. Pregnancy outcome was poorer if surgical intervention was done after >24 weeks and that, too, was done as emergency surgery. PMID- 12580155 TI - Harm reduction, public health, and human rights: smokers have a right to be informed of significant harm reduction options. AB - Public health policy needs to be assessed for effects on human rights as well as public health. Although promoting harm reduction products to cigarette smokers might lead to greater total public health harm, if the products become too popular, human rights issues also need to be considered. Avoiding, or objecting to, the fair presentation of information on effective harm reduction products to smokers to allow them to make an informed choice to reduce health risk can represent a violation of a human right - the right to information. The necessary conditions are not met for protecting public health by restricting information on certain risk reduction products. As examples, based on current evidence, smokers have a right to information on snus (Swedish moist snuff) and medicinal nicotine as harm reduction options that would reduce substantially the risk of death to individuals. Smokers also have a right to truthful information about lower-tar cigarettes that have been erroneously promoted as risk reducing. PMID- 12580156 TI - [Protease inhibitors in COPD treatment]. PMID- 12580158 TI - Tobacco harm reduction: promise and perils. AB - With the tobacco industry developing and test marketing a wide array of modified cigarettes and novel nicotine-delivery products, the era of tobacco harm reduction is upon us. Like today's new technologies, two previous generations of cigarette innovation-filtered cigarettes in the 1950s and low tar and nicotine cigarettes in the late 1960s and early 1970s were introduced to offer smokers an ostensibly less hazardous means of smoking, and therefore an alternative to quitting. Both innovations maintained cigarette sales and consequently may well have increased the morbidity and mortality toll of smoking. Will a new generation of harm reduction products improve the public's health, or will the experience of the past half-century be repeated? This paper examines the concept of tobacco harm reduction and describes the variety of methods employed in pursuit of it. Through an examination of the experience with filters and low tar and nicotine cigarettes, and an explicit consideration of today's issues and challenges, the paper focuses attention on the essential dimensions of the contemporary harm reduction debate: how science can establish whether novel products or methods will reduce risks to health for individual smokers, or at least exposures likely to influence risks; how a determination can be made as to the likely population impacts of the introduction and marketing of novel products; how health professionals and consumers can learn the potential and limits of harm reduction; and what role for governmental regulation is possible and desirable. PMID- 12580159 TI - [IgE--dependent food allergy]. PMID- 12580161 TI - SNM Highlights as history: 1978. PMID- 12580160 TI - NCI to consider adding medical x-rays to human carcinogen list. PMID- 12580162 TI - [Atopic dermatitis: pathogenesis and treatment--international principle]. PMID- 12580163 TI - [Pathogenesis of allergic rhinitis]. PMID- 12580164 TI - [New beta2-agonists in asthma and COPD]. PMID- 12580165 TI - Comments on "Evidence-based medicine in the therapy of the acute respiratory distress syndrome" by Kopp, Kuhlen, Max, Rossaint. Intensive Care Med 2002; 28:244-255. PMID- 12580166 TI - Radiation cancer analysis and low dose risk assessment: new developments and perspectives. Proceeding of the 20th LH Gray Conference. February 17-21, 2002. Ede, The Netherlands. PMID- 12580167 TI - Persistent primary peritonitis due to group A streptococcus and E. coli. PMID- 12580168 TI - Extravascular lung water and intravascular volume monitoring. PMID- 12580169 TI - Proceedings of the Contrast Media Research Symposium. Capri, Italy. October 14 18, 2001. PMID- 12580170 TI - Tracking health care costs: hospital care key cost driver in 2000. AB - This Data Bulletin is based on data from the Milliman USA Health Cost Index ($0 deductible), which is designed to reflect claims increases faced by private insurers; the Kaiser Family Foundation/Health Research and Educational Trust survey of employer-based health plans for 1999-2001; the KPMG survey of employer based plans for 1991-98; the U.S. Bureau of Labor Statistics Employment, Hours and Earnings series to track payroll costs; and Center for Studying Health System Change 2000-01 site visits (see www.hschange.org). PMID- 12580171 TI - Recycling efficiencies of C, H, O, N, S, and P elements in a Biological Life Support System based on microorganisms and higher plants. AB - MELiSSA is a microorganism based artificial ecosystem conceived as a tool for understanding the behavior of ecosystems and developing the technology for future Manned Space Missions. MELiSSA is composed of four compartments colonized by the microorganisms required by the function of this ecosystem : breakdown of waste produced by men, regeneration of atmosphere and biosynthesis of edible biomass. This paper reports the mass balance description of a Biological Life Support System composed of the MELiSSA loop and of a Higher Plant Compartment working in parallel with the photosynthetic Spirulina compartment producing edible biomass. The recycling efficiencies of the system are determined and compared for various working conditions of the MELiSSA loop with or without the HPC. PMID- 12580172 TI - The "C.E.B.A.S. MINI-MODULE": a self-sustaining closed aquatic ecosystem for spaceflight experimentation. AB - The C.E.B.A.S. MINI-MODULE is the miniaturized space flight version of the Closed Equilibrated Biological Aquatic System (C.E.B.A.S.). It fits into a large middeck locker tray and is scheduled to be flown in the STS 85 and in the NEUROLAB missions. Its volume is about 9 liters and it consists of two animal tanks, a plant cultivator, and a bacteria filter in a monolithic design. An external sensor unit is connected to a data acquisition/control unit. The system integrates its own biological life support. The CO2 exhaled by the consumers (fishes, snails, microorganisms) is assimilated by water plants (Ceratophyllum demersum) which provide them with oxygen. The products of biomass degradation and excretion (mainly ammonia ions) are converted by bacteria into nitrite and nitrate. The latter is taken up by the plants as a nitrogen source together with other ions like phosphate. The plants convert light energy into chemical energy and their illumination is regulated via the oxygen concentration in the water by the control unit. In ground laboratory tests the system exhibited biological stability up to three month. The buffer capacity of the biological filter system is high enough to eliminate the degradation products of about one half of the dead animal biomass as shown in a "crash test". A test series using the laboratory model of the flight hardware demonstrated the biological stability and technical reliability with mission-identical loading and test duration. A comprehensive biological research program is established for the C.E.B.A.S. MINI MODULE in which five German and three U.S.-American universities as well as the Russian Academy of Sciences are involved. PMID- 12580173 TI - Pigment composition and concentrations within the plant (Ceratophyllum demersum L.) component of the STS-89 C.E.B.A.S. Mini-Module spaceflight experiment. AB - The Closed Equilibrated Biological Aquatic System (C.E.B.A.S.) Mini-Module, a Space Shuttle middeck locker payload which supports a variety of aquatic inhabitants (fish, snails, plants and bacteria) in an enclosed 8.6 L chamber, was tested for its biological stability in microgravity. The aquatic plant, Ceratophyllum demersum L., was critical for the vitality and functioning of this artificial mini-ecosystem. Its photosynthetic pigment concentrations were of interest due to their light harvesting and protective functions. "Post-flight" chlorophyll and carotenoid concentrations within Ceratophyllum apical segments were directly related to the quantities of light received in the experiments, with microgravity exposure (STS-89) failing to account for any significant deviation from ground control studies. PMID- 12580174 TI - [Analgesia during birth. Tales from 1001 nights over the way back to the Garden of Eden]. PMID- 12580175 TI - [Professionally directed emergency medicine?]. PMID- 12580176 TI - [Emergency medicine -- quo vadis?!]. PMID- 12580178 TI - Layer-by-layer deposition and ordering of low-molecular-weight dye molecules for second-order nonlinear optics. PMID- 12580177 TI - Early development of fern gametophytes in microgravity. AB - Dormant spores of the fern Ceratopteris richardii were flown on Shuttle mission STS-93 to evaluate the effects of micro-g on their development and on their pattern of gene expression. Prior to flight the spores were sterilized and sown into one of two environments: (1) Microscope slides in a video-microscopy module; and (2) Petri dishes. All spores were then stored in darkness until use. Spore germination was initiated on orbit after exposure to light. For the spores on microscope slides, cell level changes were recorded through the clear spore coat of the spores by video microscopy. After their exposure to light, spores in petri dishes were frozen in orbit at four different time points during which on earth gravity fixes the polarity of their development. Spores were then stored frozen in Biological Research in Canister units until recovery on earth. The RNAs from these cells and from 1-g control cells were extracted and analyzed on earth after flight to assay changes in gene expression. Video microscopy results revealed that the germinated spores developed normally in microgravity, although the polarity of their development, which is guided by gravity on earth, was random in space. Differential Display-PCR analyses of RNA extracted from space-flown cells showed that there was about a 5% change in the pattern of gene expression between cells developing in micro-g compared to those developing on earth. PMID- 12580180 TI - [Transient neurologic symptoms following spinal anesthesia. Reply]. PMID- 12580179 TI - Plant growth during the Greenhouse II experiment on the Mir orbital station. AB - We carried out three experiments with Super Dwarf wheat in the Bulgarian/Russian growth chamber Svet (0.1 m2 growing area) on the Space Station Mir. This paper mostly describes the first of these NASA-supported trials, began on Aug. 13, 1995. Plants were sampled five times and harvested on Nov. 9 after 90 days. Equipment failures led to low irradiance (3, then 4 of 6 lamp sets failed), instances of high temperatures (ca. 37 degrees C), and sometimes excessive substrate moisture. Although plants grew for the 90 d, no wheat heads were produced. Considering the low light levels, plants were surprisingly green, but of course biomass production was low. Plants were highly disoriented (low light, mirror walls?). Fixed and dried samples and the root module were returned on the U.S. Shuttle Atlantis on Nov. 20, 1995. Samples of the substrate, a nutrient charged zeolite called Balkanine, were taken from the root module, carefully examined for roots, weighed, dried, and reweighed. The Svet control unit and the light bank were shipped to Moscow. An experiment validation test (EVT) of plant growth and experimental procedures, carried out in Moscow, was highly successful. Equipment built in Utah to measure CO2, H2O vapor, irradiance, air and leaf (IR) temperature, O2, pressure, and substrate moisture worked well in the EVT and in space. After this manuscript was first prepared, plants were grown in Mir with a new light bank and controller for 123 d in late 1996 and 39 days in 1996/1997. Plants grew exceptionally well with higher biomass production than in any previous space experiment, but the ca. 280 wheat heads that were produced in 1996 contained no seeds. Ethylene in the cabin atmosphere was responsible. PMID- 12580181 TI - [Combined spinal and epidural anesthesia (CSE) during childbirth]. PMID- 12580182 TI - Induced abnormality in Mir- and Earth grown Super Dwarf wheat. AB - Super-dwarf wheat grown on the Mir space station using the Svet "Greenhouse" exhibited morphological, metabolic and reproductive abnormalities compared with Earth-grown wheat. Of prominent importance were the abnormalities associated with reproductive ontogeny and the total absence of seed formation on Mir. Changes in the apical meristem associated with transition from the vegetative phase to floral initiation and development of the reproductive spike were all typical of 'Super-Dwarf' wheat up to the point of anthesis. Observation of ruptured anthers from the Mir-grown plants revealed what appeared to be normally developed pollen. These pollen gains, however, contained only one nuclei, while normal viable pollen is tri-nucleate. A potentially important difference in the flight experiment, compared with ground reference studies, was the presence of a high level of atmospheric ethylene (1,200 ppb). Ground studies conducted by exposing 'Super-Dwarf' wheat to ethylene just prior to anthesis resulted in manifestation of the same abnormalities observed in the space flight samples. PMID- 12580183 TI - Statement on privacy and confidentiality. PMID- 12580184 TI - Slowing Parkinson's disease progression: recent dopamine agonist trials. AB - In recent clinical trials, chronic treatment of patients with PD with pramipexole or ropinirole was associated with a slower decline of imaged striatal dopaminergic signal, compared to levodopa monotherapy. Although this could reflect slowed progression of PD, equally plausible is a pharmacologic effect on proteins that interact with the imaging radioligands. To date, there is no compelling evidence favoring dopamine agonists over levodopa; either is an appropriate choice for initial treatment of PD. PMID- 12580185 TI - Initial agonist treatment of Parkinson disease: a critique. AB - The evidence supporting initial dopamine agonist treatment of PD is reviewed. The two rationales for initial agonist treatment are reduced frequency of motor complications and possible relative neuroprotection by dopamine agonists. The basic science supporting these rationales is equivocal. The clinical evidence for advantages of initial agonist treatment is incomplete. More data are required to determine the optimal initial treatment for PD. PMID- 12580186 TI - The thrust plate prosthesis in patients with aseptic osteonecrosis of the femoral head. AB - BACKGROUND: The thrust plate prosthesis (TPP) is an implant with a metaphyseal fixation at the proximal femur that transmits the load forces of the hip onto the femoral neck. The osseous incorporation of the TPP and the adaptation of the bone to this force transmission depend on the bone quality, which is reduced to minor vitality and stability in patients with osteonecrosis of the femoral head. Depending on the etiology of the femoral head necrosis, the TPP might lead to early failures. METHODS: In a prospective study, 63 patients with 72 cementless TPP due to femoral head osteonecrosis were examined. A clinical and radiological evaluation was performed preoperatively, 3 and 6 months postoperatively, and every year thereafter. The average follow-up period was 4.8+/-1.3 years with a minimum of 3 years. The pathogenesis of femoral head necrosis included alcoholism (n=19), subsequent to renal transplantation (n=11), during cortisone therapy of other dyscrasia (n=9), preceding a polychemotherapy (n=4), diabetes (n=3), sickle cell anemia (n=1), and idiopathic osteonecrosis (n=25). RESULTS: The Harris Hip Score increased continuously from 50.0 points beyond 79.8 points after 3 months to 86.8 points within the 1st year, and subsequently remained stable at this level. Revision was necessary in six cases (8.3%). Of these, three had an aseptic loosening of the implant: 2 cases with renal transplantation and 1 of alcoholism with an extension of the necrotic area to the seating of the TPP. The other three patients showed septic implant loosenings: 2 cases with renal transplantation and 1 of alcoholism. Radiolucent lines were found in 9 cases (12.5%), mostly in zones 1 and 2 underneath the TPP. Of these, 1 with an idiopathic osteonecrosis was assessed to be radiologically loosened. The overall failure rate was 9.7%, with a proportion of 36.4% in patients with renal transplantation. Excluding this specific patient group, the failure rate was 4.9%. CONCLUSIONS: Femoral head necrosis following renal transplantation and extension of the necrotic area into the femoral neck are contraindications for TPP. Excluding these patients, the TPP shows comparable mid-term results to cementless stemmed prostheses and supplies advantages especially for younger patients, because of its metaphyseal, bone preserving fixation. However, evaluation of the clinical impact of the TPP in comparison with other cementless femoral stem systems requires long-term examinations in the future. PMID- 12580187 TI - High temperature effect on microflora of radish root-inhabited zone and nutrient solutions for radish growth. AB - The effect of high temperatures (35 and 45 degrees C) on microflora of the root zone of radish plants grown in phytotron was evaluated by the response of microorganisms from 9 indicator groups. Phytotron air temperature elevated to 35 degrees C for 20 hours caused no significant changes in qualitative and quantitative composition of the root microflora in experimental plants. By the end of the experiment, the species diversity of microflora had changed. The amount of phytopathogenic microorganisms decreased which can be interpreted as more stable co-existence of microflora with plants. The numbers of microbes from other indicator groups was in dynamic equilibrium. The plants' condition did not deteriorate either. Exposure to the temperature of 45 degrees C for 7 hours have been found to change the numbers and species diversity in the radish root zone microflora. The microorganisms were observed to increase their total numbers at the expense of certain indicator groups. Bacteria increased spore forms at the stage of spores. Colon bacillus bacteria of increased their numbers by the end of experiment by an order. By the end of experiment the roots of experiment plants had microscopic fungi from Mucor, Aspergillus, Trichoderma, Cladosporium genera. The observed changes in the microbial complex seem to be associated with the changes of root emissions and general deterioration of the plants' condition. It is suggested that the response of the microorganisms can be indicative of the condition of plants under investigation. PMID- 12580188 TI - Short-term and long-term effects of low total pressure on gas exchange rates of spinach. AB - In this study, spinach plants were grown under atmospheric and low pressure conditions with constant O2 and CO2 partial pressures, and the effects of low total pressure on gas exchange rates were investigated. CO2 assimilation and transpiration rates of spinach grown under atmospheric pressure increased after short-term exposure to low total pressure due to the enhancement of leaf conductance. However, gas exchange rates of plants grown at 25 kPa total pressure were not greater than those grown at atmospheric pressure. Stomatal pore length and width were significantly smaller in leaves grown at low total pressure. This result suggested that gas exchange rates of plants grown under low total pressure were not stimulated even with the enhancement of gas diffusion because the stomatal size and stomatal aperture decreased. PMID- 12580189 TI - A new relative referencing method for crop monitoring using chlorophyll fluorescence. AB - The measurement of plant chlorophyll fluorescence has been used for many years as a method to monitor a plant's health status. These types of methods have been mostly relegated to the laboratory. The newly developed Relative Referencing Method allows for the measurement of chlorophyll fluorescence under artificial lighting conditions. The fluorescence signal can be determined by first taking a reference signal measurement, then a second measurement with an additional fluorescence excitation source. The first signal can then be subtracted from the second and the plant's chlorophyll fluorescence due to the second lighting source can be determined. With this simple approach, a photosynthesizing plant can be monitored to detect signs of water stress. Using this approach experiments on tomato plants have shown that it was possible to detect water stress, while the plants were continuously illuminated by fluorescent lamps. This method is a promising tool for the remote monitoring of crops grown in a CELSS-type application. PMID- 12580190 TI - Biodegradation pathway of an anionic surfactant (Igepon TC-42) during recycling waste water through plant hydroponics for advanced life support during long duration space missions. AB - The degradation of an anionic surfactant (Igepon TC-42) was investigated as part of an integrated study of direct recycling of human hygiene water through hydroponic plant growth systems. Several chemical approaches were developed to characterize the degradation of Igepon and to measure the accumulation of intermediates such as fatty acids and methyl taurine. Igepon was rapidly degraded as indicated by the reduction of methylene blue active substances (MBAS) and component fatty acids. The Igepon degradation rate continued to increase over a period of several weeks following repeated daily exposure to 18 micrograms/l Igepon. The accumulation of free fatty acids and methyl taurine was also observed during decomposition of Igepon. The concentration of methyl taurine was below detection limit (0.2 nmol/ml) during the slow phase of Igepon degradation, and increased to 1-2 nmol/ml during the phase of rapid degradation. These findings support a degradation pathway involving initial hydrolysis of amide to release fatty acids and methyl taurine, and subsequent degradation of these intermediates. PMID- 12580191 TI - Performance of the CELSS Antarctic Analog Project (CAAP) crop production system. AB - Regenerative life support systems potentially offer a level of self-sufficiency and a decrease in logistics and associated costs in support of space exploration and habitation missions. Current state-of-the-art in plant-based, regenerative life support requires resources in excess of allocation proposed for candidate mission scenarios. Feasibility thresholds have been identified for candidate exploration missions. The goal of this paper is to review recent advances in performance achieved in the CELSS Antarctic Analog Project (CAAP) in light of the likely resource constraints. A prototype CAAP crop production chamber has been constructed and operated at the Ames Research Center. The chamber includes a number of unique hardware and software components focused on attempts to increase production efficiency, increase energy efficiency, and control the flow of energy and mass through the system. Both single crop, batch production and continuous cultivation of mixed crops production studies have been completed. The crop productivity as well as engineering performance of the chamber are described. For each scenario, energy required and partitioned for lighting, cooling, pumping, fans, etc. is quantified. Crop production and the resulting lighting efficiency and energy conversion efficiencies are presented. In the mixed-crop scenario, with 27 different crops under cultivation, 17 m2 of crop area provided a mean of 515 g edible biomass per day (85% of the approximate 620 g required for one person). Enhanced engineering and crop production performance achieved with the CAAP chamber, compared with current state-of-the-art, places plant-based life support systems at the threshold of feasibility. PMID- 12580192 TI - Third International Conference on Oxygen/Nitrogen Radicals: cell injury and disease. Morgantown, West Virginia, June 1-5, 2002. PMID- 12580193 TI - Would Healing: oxygen and emerging therapeutics. Columbus, Ohio, September 12-15, 2002. PMID- 12580195 TI - Copy cost questions require closer look. PMID- 12580194 TI - Initial closed operation of the CELSS Test Facility Engineering Development Unit. AB - As part of the NASA Advanced Life Support Flight Program, a Controlled Ecological Life Support System (CELSS) Test Facility Engineering Development Unit has been constructed and is undergoing initial operational testing at NASA Ames Research Center. The Engineering Development Unit (EDU) is a tightly closed, stringently controlled, ground-based testbed which provides a broad range of environmental conditions under which a variety of CELSS higher plant crops can be grown. Although the EDU was developed primarily to provide near-term engineering data and a realistic determination of the subsystem and system requirements necessary for the fabrication of a comparable flight unit, the EDU has also provided a means to evaluate plant crop productivity and physiology under controlled conditions. This paper describes the initial closed operational testing of the EDU, with emphasis on the hardware performance capabilities. Measured performance data during a 28-day closed operation period are compared with the specified functional requirements, and an example of inferring crop growth parameters from the test data is presented. Plans for future science and technology testing are also discussed. PMID- 12580197 TI - Perfecting the communication process. PMID- 12580196 TI - "Dispensing death, desiring death" by Annette Street and David Kissane: a commentary. PMID- 12580198 TI - Of euphemisms and euthanasia: the language games of the Nazi doctors and some implications for the modern euthanasia movement. AB - Euphemisms are place-holders for important concepts. They may disguise a practice which one might abhor if it were given another name. In Nazi Germany during World War II, euphemisms were used to desensitize physicians and society to the horrors of a program of euthanasia. This article examines some of the euphemisms used by the Nazi physicians to redefine medicalized killing, compares the Nazi language games with those of contemporary proponents of medicalized killing, and concludes that the consistent application of euphemisms for medicalized killing significantly weakens arguments against assisted killing. PMID- 12580199 TI - Kevorkian, Martha Wichorek and us: a personal account. AB - In this short article, the authors describe their attempt to do suicide prevention with a patient that ultimately died as the result of a physician assisted suicide. Autopsy revealed no sign of physical disease but the patient's letters indicate a preoccupation with independence as the definition of life, and conviction that people who lose independence are no longer alive. PMID- 12580200 TI - Michigan versus Kevorkian. AB - Our personal reflections on the Michigan versus Kevorkian trial highlight the following issues: 1) the switch from physician-assisted suicide to euthanasia, 2) the television showing of the death, 3) the dropping of the prosecution of the charge of physician-assisted suicide, 4) Kevorkian serving as his own defense attorney, trying to argue that ALS was a secondary cause of Thomas Youk's death, 5) Kevorkian's attempt to employ a logical syllogism to demonstrate that euthanasia need not be murder, 6) Kevorkian's initial reference to the civil rights tradition but sudden change to the medical analogy of Nazi medicine: a final solution, 7) the insistence of Kevorkian on "all or nothing" sentencing, 8) the irony of Kevorkian being finally convicted by a prosecutor who was elected on a platform of not prosecuting Kevorkian, 9) Kevorkian hiring a lawyer after the verdict is in, and 10) Kevorkian's threat to starve himself to death if sent to prison. PMID- 12580201 TI - Physician, hate thyself: comments on the Kevorkian tapes. PMID- 12580202 TI - Summary. PMID- 12580203 TI - How bioterrorism became an HIM issue. PMID- 12580204 TI - A second generation technique for evaluating accuracy and reliability of second generation pulse oximeters. PMID- 12580205 TI - Masimo signal extraction pulse oximetry. AB - OBJECTIVE: To describe a new pulse oximetry technology and measurement paradigm developed by Masimo Corporation. INTRODUCTION: Patient motion, poor tissue perfusion, excessive ambient light, and electrosurgical unit interference reduce conventional pulse oximeter (CPO) measurement integrity. Patient motion frequently generates erroneous pulse oximetry values for saturation and pulse rate. Motion-induced measurement error is due in part to widespread implementation of a theoretical pulse oximetry model which assumes that arterial blood is the only light-absorbing pulsatile component in the optical path. METHODS: Masimo Signal Extraction Technology (SET) pulse oximetry begins with conventional red and infrared photoplethysmographic signals, and then employs a constellation of advanced techniques including radiofrequency and light-shielded optical sensors, digital signal processing, and adaptive filtration, to measure SpO2 accurately during challenging clinical conditions. In contrast to CPO which calculates O2 saturation from the ratio of transmitted pulsatile red and infrared light, Masimo SET pulse oximetry uses a new conceptual model of light absorption for pulse oximetry and employs the discrete saturation transform (DST) to isolate individual "saturation components" in the optical pathway. Typically, when the tissue under analysis is stationary, only the single saturation component produced by pulsatile arterial blood is present. In contrast, during patient motion, movement of non-arterial components (for example, venous blood) can be identified as additional saturation components (with a lower O2 saturation). When conditions of the Masimo model are met, the saturation component corresponding to the highest O2 saturation is reported by the instrument as SpO2. CONCLUSION: The technological strategies implemented in Masimo SET pulse oximetry effectively permit continuous monitoring of SpO2 during challenging clinical conditions of motion and poor tissue perfusion. PMID- 12580207 TI - Echocardiography: a brief description. PMID- 12580206 TI - Specific elements of a new hemodynamics display improves the performance of anesthesiologists. AB - OBJECTIVE: We tested the hypothesis that a monitoring display proposed by Blike et al. improves the performance of anesthesiologists. We measured the performance of anesthesiologists using the new display and compared it to their performance with a traditional display. We studied three different displays on how they affected recognition and differentiation of five etiologies of shock-anaphylaxis, bradycardia, hypovolemia, ischemia and pulmonary embolus. METHODS: The participants monitored heart rate, systemic arterial and pulmonary blood pressure, central venous pressure, and cardiac output during five shock states and five non-shock states. The resulting 10 data sets made up ten decision screens, which we presented randomly on a computer monitor to the subjects in one of three different formats (a Single Sensor Single Indicator (SSSI) Numeric display; an Object display; and an Object Minus Shapes display). Subjects used soft-buttons on a computer touch-screen monitor to: a) advance to the next display; b) differentiate a non-shock state from a shock state; and, c) select the etiology of shock state represented by the display (Figure 4). The internal clock and memory of the computer made the collection of data automatic. RESULTS: The subjects recognized a problem more rapidly with the help of a graphical "pointer on a reference scale" in both Object displays, but their accuracy had not improved in comparison to the SSSI Numeric display. The shape of the Object display improved performance of etiology determination compared to the Object Minus Shapes display and SSSI Numeric display. Testing (10 trials) was completed in less than 45 minutes. CONCLUSIONS: The new display with "emergent features" can improve the diagnostic performance of clinicians. PMID- 12580208 TI - A patient with a ventricular wall motion abnormality. The first in a series of patients demonstrating the clinical value of transesophageal echocardiography. PMID- 12580209 TI - Sonic vibrational analysis provides continuous measurement of arterial properties. AB - OBJECTIVE: We describe a new technology for measuring artery mechanical properties, called Sonic Vibrational Analysis (SVA). We utilize SVA to study the changes in radial artery smooth muscle tone caused by intravenous infusion of vasoactive agents. METHODS: Six healthy volunteers were monitored with a radial intra-arterial catheter and an SVA sensor during progressively increasing doses of nitroglycerin (NTG), phenylephrine, sodium nitroprusside (SNP), dobutamine, and nicardipine. In SVA, the propagation velocity of an audio-frequency vibration is measured over a short segment of the radial artery. The measurement has sufficient temporal resolution to track the continuous changes in arterial properties that occur due to the natural blood pressure pulse. RESULTS: Coupled with the measurement of radial blood pressure, SVA allowed determination of the physiological/mechanical state of the artery within a single cardiac cycle. NTG, SNP, and phenylephrine caused significant changes in both blood pressure and the physiological state of the radial artery. Nicardipine and dobutamine altered blood pressure without change in the state of the radial artery. CONCLUSIONS: The current results are consistent with previous studies of the effects of vasoactive agents on the radial artery. SVA is non-invasive, continuous, localized to a well defined section of artery, and suitable for the collection of large volumes of time-resolved data in a laboratory or clinical setting. PMID- 12580210 TI - Determining the artifact sensitivity of recent pulse oximeters during laboratory benchmarking. AB - This study aimed to investigate and compare the performance of the algorithms contained in the newest generation of pulse oximeters (Masimo SET in IVY2000, Nellcor Oxismart N-3000, Agilent M3 rev. B) against a traditional pulse oximeter (Agilent CMS rel. A.0). The benchmark was performed in an efficient and reproducible way in the laboratory environment using patient signal recordings complemented by a two-hand volunteer motion test. A novel method of creating artifact/reference signal pairs from a clinical database, the noise-mix composition (NMC), was developed. The new method enabled the simulation of critical clinical situations in a more realistic way than the usual two hand volunteer studies. An advantage of the laboratory tests over live clinical studies was that a continuous saturation reference was available, allowing accurate on-going determination of the SpO2 error. A new quantitative performance measure, the non-performance index (NPI), was developed and applied to the benchmark results. It covers the 3 performance aspects of a pulse oximeter: (1) SpO2 accuracy, (2) pulse rate accuracy and (3) drop out times. These factors were weighted according to clinical importance determined by a survey. During the restricted conditions of steady state and forced motion test on healthy volunteers Masimo/Ivy's pulse oximeter performed best with a 2.6 fold improvement over the conventional technology. Clear improvements were also found for Agilent's M3 (1.6 fold) and Nellcor's N-3000 (1.6 fold). In contrast, the clinically oriented NMC study yielded the best performance improvement--as measured in NPI numbers--for Agilent's M3 rev. B (1.6 fold) and due to more frequent SpO2 errors only 1.5 for Masimo and 1.3 for N-3000. A large difference was found for the dropout rate: the lowest was achieved by Masimo (3.0% of total time), the largest by Nellcor N-3000 (24.1% of total time), a factor which was rated high by clinicians. Very pronounced improvements (between 2.3 and 3.4 fold) on all of the newer devices were found for the pulse rate. The NMC turned out to be a very useful tool for generating a standard signal set for algorithm development and benchmarking purposes that eliminates repetitive clinical testing in early stages. The applicability of its results needs confirmation by clinical live studies. PMID- 12580211 TI - Does monitoring of post-tetanic count prevent alarms of airway pressure or visible muscle activity during intratracheal jet ventilation? A prospective study with five different neuromuscular blocking agents. AB - OBJECTIVE: Profound neuromuscular block (NMB) quantified by post-tetanic count (PTC) may prevent all muscle activity during anesthesia. We studied whether monitoring of PTC prevents airway pressure alarms or visible movements of the vocal cords and the abdomen during endolaryngeal procedures (ELPs). METHODS: In this prospective, double blind, study 50 healthy (ASA 1-3) patients scheduled for ELPs were randomized into five groups: atracurium, mivacurium, rocuronium, vecuronium and succinylcholine. During alfentanil-propofol anaesthesia, profound NMB was controlled by monitoring the PTC (target level PTC 0-2, 50 Hz tetanic stimulation) of the adductor pollicis muscle. The muscle relaxants were administered using bolus dosing in all groups but in the succinylcholine group. The early signs of recovery of NMB to be observed were: 1) airway pressure alarms, 2) movements of vocal cords on the laryngeal video monitor and 3) movements of the abdomen. The inference was based on 90% confidence interval tests. RESULTS: During 50 ELP:s following signs of early recovery of NMB were recognized: 2 alarms of airway pressure, 16 laryngeal movements and 11 movements of the abdomen. The proportion of airway pressure alarms was significantly lower than proportion of all detectable movements (95% confidence interval analysis). Twelve of the movements were recorded at PTC zero level. The signs of early recovery of NMB were detected in all groups. CONCLUSIONS: PTC-monitoring following 50 Hz stimulation does not ensure total inactivity of muscles during alfentanil-propofol anesthesia, regardless which relaxant has been chosen. During ELPs, simultaneous observation of the vocal cords and the abdomen is more sensitive in detecting early recovery of NMB compared to our method of airway pressure monitoring. PMID- 12580212 TI - The effect of single twitch and train-of-four stimulation on twitch forces during stable neuromuscular block. AB - OBJECTIVE: We investigated whether the response to a single twitch (ST) stimulus or the first response (T1) to a train-of-four (TOF; 4 stimuli at 2 Hz) stimulus following a stimulus interval of 10 s (i.e., the time between two consecutive ST or TOF stimuli) is influenced by the preceding stimulus in the presence of a stable 50% neuromuscular block. In addition, we determined whether ST and TOF stimulation yield different results under these circumstances. METHODS: Twitch forces were measured in both tibialis anterior muscles of six cats. In the presence of a stable 50% neuromuscular block the stimulation pattern (ST or TOF) or stimulus interval (3.3, 10 or 30 s) was varied every 30 min. A linear mixed model was used for statistical analysis. RESULTS: ST forces with a stimulus interval of 3.3 s were 10.3% (95% CI: 7.3-13.3%) smaller than those with a stimulus interval of 10 s. For T1 forces this effect was 15.2% (95% CI: 12 18.4%). There was no significant difference between twitch forces with stimulus intervals of 30 and 10 s. For a stimulus interval of 3.3 s the ST forces exceeded the T1 forces by 7.6% (95% CI: 4.4-10.8%); no significant differences were found between the ST and T1 forces for stimulus intervals of 10 and 30 s. CONCLUSIONS: The ST or T1 force during stimulation with a stimulus interval of 10 s or more during a stable 50% neuromuscular block in the tibialis anterior muscle of the cat is not affected by the preceding stimulus. In addition, ST and T1 forces do not differ when employing a stimulus interval of 10 s or more under these circumstances. Our results thus indicate that the known differences between ST and T1 forces after a bolus injection of a muscle relaxant can not be explained by differences in acetylcholine release when the stimulus interval exceeds 10 s. PMID- 12580213 TI - Monitoring of isoflurane and desflurane breakdown: interfering gases and infrared detection. AB - OBJECTIVE: The reaction of isoflurane, enflurane or desflurane with dried CO2 absorbents produces carbon monixide (CO), a highly toxic gas which cannot be detected by gas monitors typically available in the operating room. Trifluoromethane (CHF3) is produced along with CO when this reaction occurs with isoflurane and desflurane, and can be detected by gas monitors. This study will determine the ability of a modified SAM module (Smart Anesthesia Multigas Module, GE/Marquette Medical Systems, Milwaukee, WI) to identify the presence of CHF3, and provide a clinically useful indirect warning of CO production. METHODS: Isoflurane (1.5%) and desflurane (7.5%) were reacted under clinical conditions with desiccated absorbents resulting in CO production. CO and CHF3 concentrations were measured using gas chromatography. The CHF3 concentrations measured by a modified SAM monitor were compared with the measurements obtained by gas chromatography. Alarm limits set on the SAM monitor were used to warn of the presence of CHF3. RESULTS: A concentration of 0.25% CHF3, as measured by the SAM monitor, corresponds to an average CO concentration of 780 ppm for isoflurane and 1700 ppm for desflurane. Lowering the threshold to 0.05% CHF3 would result in an average CO concentration of 155 ppm CO for isoflurane and 345 ppm CO for desflurane. CONCLUSIONS: We have shown that the SAM module is capable of measuring CHF3 due to anesthetic breakdown. With appropriate changes in the display programming and reference cell spectra the monitor would be able to provide an early warning of CO exposure, although the amount of CO would not be reported. PMID- 12580214 TI - A proposed new set of alarm sounds which satisfy standards and rationale to encode source information. PMID- 12580215 TI - Arterial-line monitoring system simulation. AB - OBJECTIVE: We sought to improve the realism of our patient simulation environment by developing a simulation of the arterial-line monitoring system. Properties of the system we wished to depict were: electro-mechanical delay between ECG and radial artery pressure, beat to beat amplitude variability and respiratory variation, realistic looking pulse pressure in hypertensive and hypotensive states, a functional link to the stopcock and transducer flush, and filtering characteristics of the measurement system. METHODS: A standard clinical pressure transducer and stopcock were modified to provide data about their state to a personal computer. A software program was written to modify the arterial pressure waveform from a patient simulator according to the pressure transducer and stopcock state as well as user settings to produce a new waveform. RESULTS: All of the desired improvements in the realism of the arterial waveform were implemented. CONCLUSIONS: The realism of scenarios using the patient simulator is enhanced by having the arterial-line monitoring system more accurately simulated. PMID- 12580216 TI - Atelectasis formation during anesthesia: causes and measures to prevent it. AB - Pulmonary gas exchange is regularly impaired during general anaesthesia with mechanical ventilation. This results in decreased oxygenation of blood. A major cause is collapse of lung tissue (atelectasis), which can be demonstrated by computed tomography but not by conventional chest x-ray. Collapsed lung tissue is present in 90% of all subjects, both during spontaneous breathing and after muscle paralysis, and whether intravenous or inhalational anaesthetics are used. There is a correlation between the amount of atelectasis and pulmonary shunt. Shunt does not increase with age. In obese patients, larger atelectatic areas are present than in lean ones. Finally, patients with chronic obstructive lung disease may show less or even no atelectasis. There are different procedures that can be used in order to prevent atelectasis or to reopen collapsed lung tissue. The application of positive end-expiratory pressure (PEEP) has been tested in several studies. On the average, arterial oxygenation does not improve markedly, and atelectasis may persist. Further, reopened lung units re-collapse rapidly after discontinuation of PEEP. Inflation of the lungs to an airway pressure of 40 cm H2O, maintained for 7-8 seconds (recruitment or "vital capacity" manoeuvre), re-expands all previously collapsed lung tissue. During induction of anaesthesia, the use of a gas mixture, that includes a poorly absorbed gas such as nitrogen, may prevent the early formation of atelectasis. During ongoing anaesthesia, pulmonary collapse reappears slowly if a low fraction of oxygen in nitrogen is used for the ventilation of the lungs after a previous VC-manoeuvre. On the other hand, ventilation of the lungs with pure oxygen results in a rapid reappearance of atelectasis. Thus, ventilation during anaesthesia should be done if possible with a moderate fraction of inspired oxygen (FIO2, e.g. 0.3-0.4). Alternatively, if the lungs are ventilated with a high inspiratory fraction of oxygen, the use of PEEP may be considered. In summary, atelectasis is present in most humans during anaesthesia and is a major cause of impaired oxygenation. Avoiding high fractions of oxygen in inspired gas during induction and maintenance of anaesthesia may prevent formation of atelectasis. Finally, intermittent "vital capacity"-manoeuvres together with PEEP reduces the amount of atelectasis and pulmonary shunt. PMID- 12580217 TI - Discriminating between the effect of shunt and reduced VA/Q on arterial oxygen saturation is particularly useful in clinical practice. AB - There is an extensive literature on methods for discriminating between an increased shunt and a reduced ratio of ventilation to perfusion. In this review we prefer the terms "VA/Q" and "reduced or low VA/Q" rather than "V/Q inequality" to refer to the effects on arterial oxygenation of reducing V/Q below 0.8 to about 0.1. Almost without exception the conventional methods for measuring shunt and reduced VA/Q are invasive as well as technically complex. For most clinicians who are dealing with a hypoxemic patient the relevance of these entities is not so obvious as to justify the time and difficulty in either understanding or measuring them. However this review shows that, while an increased shunt and a decreased VA/Q both reduce arterial oxygen saturation (SaO2) at a particular inspired oxygen concentration (PIO2), the effect of shunt and reduced VA/Q have important clinical differences on the relationship between PIO2 and SaO2. The review also outlines a simple non-invasive method for measuring shunt and reduced VA/Q which illustrates the value of discriminating between them in clinical practice. PMID- 12580218 TI - Non-steady state monitoring by respiratory gas exchange. AB - Traditionally, the study of CO2 and O2 kinetics in the body has been mostly confined to equilibrium conditions. However, the peri-anesthesia period and the critical care arena often involve conditions of non-steady state. The detection and explanation of CO2 kinetics during non-steady state pathophysiology have required the development of new methodologies, including the CO2 expirogram, average alveolar expired PCO2, and CO2 volume exhaled per breath. Several clinically relevant examples of non-steady state CO2 kinetics perturbations are examined, including abrupt decrease in cardiac output, application of positive end-expiratory pressure during mechanical ventilation, and occurrence of pulmonary embolism. The lesser known area of non-steady state O2 kinetics is introduced, including the measurement of pulmonary O2 uptake per breath. Future directions include the study of the respiratory quotient per breath, where the anaerobic threshold during anesthesia is identified by increasing respiratory quotient. PMID- 12580219 TI - Partial CO2 rebreathing indirect Fick technique for non-invasive measurement of cardiac output. AB - OBJECTIVE: Evaluation in animals of a non-invasive and continuous cardiac output monitoring system based on partial carbon-dioxide (CO2) rebreathing indirect Fick technique. METHODS: We have developed a non-invasive cardiac output (NICO) monitoring system, based on the partial rebreathing method. The partial rebreathing technique employs a differential form of the Fick equation for calculating cardiac output (QT) using non-invasive measurements. Changes in CO2 elimination (deltaVCO2) and partial pressure of end-tidal CO2 (deltaPETCO2) in response to a brief period of partial rebreathing are used to measure pulmonary capillary blood flow (Q(PCBF)). A non-invasive estimate of anatomic and intrapulmonary shunt fraction (Q(S)/Q(T)), based on oxygen saturation from pulse oximetry (SpO2) and inspired oxygen concentration (FIO2), is added to compute total cardiac output [Q(T) = Q(PCBF)/(1 - Q(S)/Q(T))]. The performance of the NICO was compared with iced 5% dextrose bolus thermodilution cardiac output (TDco) measurements in 6 dogs. Cardiac output was varied using dobutamine, and halothane, and by clamping of the inferior vena cava. Two hundred and forty-six (n = 246) paired measurements of TDco and NICO over a range of cardiac outputs (TDco range = 0.60-8.87 l/min) were compared using Bland-Altman analysis and weighted correlation coefficient. RESULTS: The Bland-Altman technique yielded a NICO precision of +/- 0.70 l/min (13.8%) with a mean bias of -0.07 l/min (-1.4%) compared to TDco. The weighted correlation coefficient between TDco and NICO values was: r = 0.93 (n = 246). CONCLUSION: The partial CO2 rebreathing technique for measurement of cardiac output is non-invasive, automated, and based on the well accepted Fick principle. The limits of agreement between NICO and TDco is within the recommended value for NICO to be a clinically acceptable method for cardiac output measurement. The results of this canine study show that NICO performed as well, and in some cases better, than other currently available non invasive cardiac output techniques over a wide range of cardiac outputs. PMID- 12580220 TI - Pulmonary air embolism. AB - Pulmonary air embolism is a well-known consequence of surgery, trauma, diving, and aviation. This article reviews the physiological effects, means of detection and methods of prevention and treatment of pulmonary air embolism. The primary physiological effects are elevated pulmonary artery pressures, increased ventilation-perfusion inhomogeneity, and right ventricular failure. The degree of physiological impairment depends on the volume of gas entrained, the rate of entrainment, the type of gas entrained, and the position of the patient when the embolism occurs. Transesophageal echocardiography is the most sensitive method of detection, but it is invasive. Precordial Doppler ultrasound is almost as sensitive and poses no risk to the patient. End-tidal carbon dioxide monitoring is used on all patients and is a moderately sensitive method of detection, which is useful during surgeries that have a low incidence of air embolism. For high risk procedures, precordial Doppler ultrasound and a multi-orifice right heart catheter should be used to detect and treat pulmonary air embolism. Prevention measures include volume expansion, careful positioning, positive end-expiratory pressure, military anti-shock trousers, and jugular venous compression. Treatment of pulmonary air embolism includes flooding the surgical site with saline, controlling sites of air entry, repositioning the patient with the surgical site below the right atrium, aspiration of air from a central venous catheter, cessation of inhaled nitrous oxide, and resuscitation with oxygen, intravenous fluids, and inotropic agents. Some hypotheses on the effects of air in the pulmonary vasculature and investigational treatment options are discussed. PMID- 12580221 TI - Monitoring alveolar epithelial function in acute lung injury. AB - OBJECTIVE: Identification of humoral markers of acute lung injury may lead to insights into pathologic mechanisms. In addition, specific markers may be useful for predicting development of acute respiratory distress syndrome (ARDS) or for assessing prognosis. Ultimately, studies of lung injury markers may help define interventions that prevent or moderate ARDS. The alveolar epithelium is important both for the integrity of the blood-gas barrier and for repair of the barrier after lung injury. This article reviews markers that derive from or relate to the alveolar epithelium and that might be used for monitoring alveolar epithelial function in acute lung injury. Surfactant apoproteins may be important markers of injury or for prognosis. Levels of surfactant apoprotein A (SP-A) fall 50-75% in patients with severe lung injury compared to normal patients. Serum levels of SP A in patients dying of acute respiratory distress syndrome are double serum levels of survivors. PMID- 12580223 TI - Noninvasive ventilation for acute respiratory failure. AB - Noninvasive ventilation refers to any form of ventilatory support applied without the use of an endotracheal tube. It offers the potential to provide primary treatment for acute respiratory failure while avoiding complications associated with mechanical ventilation with endotracheal intubation. Noninvasive ventilation has been most commonly studied in hypercapnic respiratory failure. A review of randomized, controlled studies shows mixed results and methodologic limitations affect the interpretation of current evidence. Patient selection is clearly the most important issue in considering noninvasive ventilation for acute respiratory failure. Unfortunately, patients who benefit from noninvasive ventilation represent only a minority of the total group with any one disease, and thus it is difficult to make broad conclusions concerning applicability of this treatment modality. Future studies are needed to focus on determining the specific patient populations who will benefit the most, evaluating the optimal ventilatory mode and mask for providing noninvasive ventilation, and clarifying its impact on clinical outcomes. PMID- 12580222 TI - Inhaled nitric oxide and pulmonary vasoreactivity. AB - Inhaled nitric oxide is a ubiquitous molecule which is produced endogenously and is also found in air pollution and in cigarette smoke. After describing the chemistry of NO, we review its history from the first description in 1980 to the current clinical indications. The biosynthesis of NO, its effects on pulmonary vasoreactivity, and the administration of inhaled NO will be described. The indications, uses, and side effects of inhaled NO are discussed with an emphasis on withdrawal of NO therapy, specifically the "rebound" phenomenon. Possible drug interactions are listed. Inhaled nitric oxide is here to stay, and future studies will provide more information on its therapeutic dose, duration and potential toxicity. PMID- 12580224 TI - Monitoring ventilator weaning--predictors of success. AB - Improving the prediction of successful ventilator weaning and extubation is a goal that all Intensivists and perioperative physicians strive for. The successful wean and extubation of ventilated patients decreases hospital length of stay and associated costs, but more importantly it also reduces patient morbidity and mortality. This review evaluates traditional and novel indices used in the assessment for ventilatory wean readiness. Novel equipment such as the Bicore pulmonary monitor and the CO2 SMO Plus are now available on the market to assess and monitor ventilator weaning and may offer some value in this process. We also review the non-respiratory factors affecting weaning and the role of the bedside nurse and respiratory therapist. Resolution of the pulmonary compromise and an understanding of respiratory physiology, used in conjunction with monitored indices of weaning parameters in a consistent fashion will continue to improve our success rates of ventilator weaning and extubation. PMID- 12580225 TI - So you want to be a pulmonary mechanic: a clinical guide. AB - This review provides a practical clinical guide to the measurement of pulmonary mechanics. Although these measurements are now commonly available in a variety of clinical settings, there is considerable confusion regarding their interpretation and significance. A basic understanding of the principles involved will help prevent the misuse of this important information. PMID- 12580226 TI - Assessment and monitoring of flow limitation and other parameters from flow/volume loops. AB - Flow/volume (F/V) spirometry is routinely used for assessing the type and severity of lung disease. Forced vital capacity (FVC) and timed vital capacity (FEV1) provide the best estimates of airflow obstruction in patients with asthma, chronic obstructive pulmonary disease (COPD) and emphysema. Computerized spirometers are now available for early home recognition of asthma exacerbation in high risk patients with severe persistent disease, and for recognition of either infection or rejection in lung transplant patients. Patients with severe COPD may exhibit expiratory flow limitation (EFL) on tidal volume (VT) expiratory F/V (VTF/V) curves, either with or without applying negative expiratory pressure (NEP). EFL results in dynamic hyperinflation and persistently raised alveolar pressure or intrinsic PEEP (PEEPi). Hyperinflation and raised PEEPi greatly enhance dyspnea with exertion through the added work of the threshold load needed to overcome raised pleural pressure. Esophageal (pleural) pressure monitoring may be added to VTF/V loops for assessing the severity of PEEPi: 1) to optimize assisted ventilation by mask or via endotracheal tube with high inspiratory flow rates to lower I:E ratio, and 2) to assess the efficacy of either pressure support ventilation (PSV) or low level extrinsic PEEP in reducing the threshold load of PEEPi. Intraoperative tidal volume F/V loops can also be used to document the efficacy of emphysema lung volume reduction surgery (LVRS) via disappearance of EFL. Finally, the mechanism of ventilatory constraint can be identified with the use of exercise tidal volume F/V loops referenced to maximum F/V loops and static lung volumes. Patients with severe COPD show inspiratory F/V loops approaching 95% of total lung capacity, and flow limitation over the entire expiratory F/V curve during light levels of exercise. Surprisingly, patients with a history of congestive heart failure may lower lung volume towards residual volume during exercise, thereby reducing airway diameter and inducing expiratory flow limitation. PMID- 12580227 TI - Non-invasive imaging of regional lung function using x-ray computed tomography. AB - The use of imaging technologies has progressed beyond the depiction of anatomic abnormalities to providing non-invasive regional structure and functional information in intact subjects. These data are particularly valuable in studies of the lung, since lung disease is heterogeneous and significant loss of function may occur before it is detectable by traditional whole lung measurements such as oxygenation, compliance, or spirometry. While many imaging modalities are available, X-ray computed tomography (CT) is emerging as the preferred method for imaging the lung because of its widespread availability, resolution, high signal/noise ratio for lung tissue, and speed. Utilizing the quantitative density and dimensional information available from conventional CT images, it is possible to measure whole and regional lung volumes, distribution of lung aeration and recruitment behavior under various clinical conditions and interventions, and important regional mechanical properties. In addition, using the radiodense gas xenon (Xe) as a contrast agent, regional ventilation or gas transport may also be obtained. This communication will review recent advances in CT based techniques for the measurement of regional lung function. PMID- 12580228 TI - HRCT imaging of airway responsiveness: effects of anesthetics. AB - The noninvasive imaging method, high resolution computed tomography (HRCT), has been developed in animal models and applied to humans with obstructive lung disease for assessing regional and individual airway responsiveness. The ability to directly view airway responses during provocations such as tracheal intubation in an asthmatic could greatly enhance our understanding and treatment of airway hyperresponsiveness. HRCT uses increased kilovoltage peak (kVp) and milliamperage (mAs) settings, thin slices, high spacial frequency reconstruction algorithms, and small fields of view to resolve structures as small as 200 microm. Therefore, airways as small as 1-2 mm in diameter can be viewed and measured. HRCT is a more sensitive technique for resolving airway caliber changes than clinical or research methods of pulmonary function tests. HRCT allows direct in vivo measurement of airway responsiveness to pharmacological and physiological stress that induces bronchoconstriction or bronchodilation. Using HRCT, we are able to measure airway dilation at baseline airway tone with inhalation anesthetics, differentiate the bronchodilating properties of inhalational agents in airways with tone, assess bronchodilating agents commonly used as premedications prior to anesthesia, and measure airway heterogeneity at baseline tone and their response to a variety of stimuli. This ability of HRCT to measure airway caliber and response heterogeneity in vivo noninvasively will dramatically improve our understanding of pulmonary physiology in general and the effects of anesthetics on the airways specifically. PMID- 12580230 TI - Teaching respiratory physiology: clinical correlation with a human patient simulator. AB - In recent years students have increasingly objected to laboratory exercises involving animal subjects. We have replaced the valuable animal experiments with demonstrations using a full-scale human patient simulator. In small groups first year medical students observe realistic clinical situations such as opioid induced hypoventilation, pneumothorax, and pulmonary edema. Students obtain information through physical examination, arterial blood gas analysis and chest radiography. They practice interventions such as providing supplemental oxygen and mask ventilation, monitor the results, and develop a basic differential diagnosis and treatment plan. We utilize the clinical context to review fundamental concepts of respiratory physiology including the alveolar air equation and oxyhemoglobin dissociation curve. The students give these laboratory exercises uniformly superior evaluations. PMID- 12580229 TI - Effects of high thoracic epidural anesthesia and local anesthetics on bronchial hyperreactivity. AB - Bronchial hyperreactivity can cause life threatening bronchospasm after airway irritation. Therefore, endotracheal intubation is avoided in asthmatics when feasible. High thoracic epidural anesthesia can be used to avoid endotracheal intubation and offers less postoperative pulmonary complications when compared to systemic postoperative analgesia. However, there are concerns that it might also cause impaired ventilation by extended motor blockade, increased airway resistance, and increased bronchial reactivity because of pulmonary sympathicolysis. Nevertheless, high thoracic epidural anesthesia causes only a slight decrease in vital capacity and neither an increase in airway resistance nor increased bronchial reactivity. In fact, it causes a decrease in bronchial reactivity in patients with bronchial hyperreactivity mostly due to the systemic effect of the local anesthetic. The attenuation of bronchial hyperreactivity can be shown as a dose dependent effect of lidocaine and bupivacaine. The intravenous effect of lidocaine is comparable to the effect of a moderate dose of salbutamol and leads to an additive effect when both drugs are used in combination. Overall, high thoracic epidural anesthesia can be used safely in patients with bronchial hyperreactivity and intravenous administration of lidocaine (1.5-2.0 mg x kg(-1)) can be used as a prophylactic treatment prior to airway instrumentation. PMID- 12580231 TI - Comparison of non-invasive blood pressure measurements on the arm and calf during cesarean delivery. AB - OBJECTIVE: Shivering may occur in 75% of women undergoing spinal anesthesia for cesarean delivery and may render an automated noninvasive blood pressure (ANIBP) device incapable of determining blood pressurc (BP). When patients shiver under spinal anesthesia, the lower extremities do not exhibit the same involuntary muscle movements as do the upper extremities. This study was undertaken to determine if a correlation exists between ANIBP measurements in the arm and calf of women undergoing cesarean delivery under spinal anesthesia. METHODS: We enrolled 73 women in this blinded, prospective study. Simultaneous arm and calf BP were measured with an ANIBP and differences between the two were determined. RESULTS: We found significant differences between the average difference in systolic and in diastolic BP, no significant difference between the average mean BP, and a tendency for the systolic BP to be higher and the diastolic BP to be lower in the calf than in the arm; however, there was a large degree of variability among patients. CONCLUSION: We conclude that there is a poor correlation between the BP measured by an ANIBP on the calf and one on the arm. In the parturient undergoing cesarean section, lower extremity BP as measured by an ANIBP does not correlate with the arm ANIBP and should not be used to assure fetal well being. PMID- 12580232 TI - Accuracy of deadspace free ventilatory measurements for lung function testing in ventilated newborns: a simulation study. AB - OBJECTIVE: A deadspace free method based on simultaneous ventilatory measurements in the inspiratory and expiratory limb of the ventilator circuit was compared to the conventional endotracheal method where the flow is measured between ETT and Y Piece. The aim of our study was to find out how the arrangement of this setup affects the measuring accuracy of 1) the ventilatory and 2) the lung mechanical parameters by means of a computer simulation. METHOD: The system consisting of ventilator tubes and lung was described in state space and the flow signals of endotracheal method, of deadspace free method and the pressure at the Y-piece were simulated in the time domain. To investigate the influence of the position of the pneumotachographs (PNTs) in the ventilator circuit on measuring accuracy, the distance d0 of the PNT from the Y-piece was varied between 0 and 900 mm. The respiratory compliance C, resistance R and inertance I were calculated by least square method using the simulated flow and pressure signals of both methods. RESULTS: Compared to the endotracheal method, with increasing d0, the tidal volume measured with the deadspace free method rose linearly, depending on the ratio between the compliance of the ventilator tubes to the respiratory compliance. The differences of C and R for both methods were acceptable (< 10%) if the distance between each PNT and the y-piece didn't exceed 200 mm and the shorter do the higher the measuring accuracy. The inertance could not be measured by this method with satisfactory accuracy if d0 was higher than 100 mm. IN CONCLUSION: The dead space free method can be used for accurate ventilatory measurements during mechanical ventilation. However, for lung mechanic measurements in very low birth weight infants the position of the PNTs must be as short as possible. PMID- 12580233 TI - Influence of tidal volume and thoraco-abdominal separation on the respiratory induced variation of the photoplethysmogram. AB - OBJECTIVE: The present study was aimed at determining the relative influences of tidal volume and thoraco-abdominal separation (relative thoracic and abdominal contribution to the tidal volume) on the respiratory induced intensity variation (RIIV) of the photoplethysmographic signal. The effects were studied in two body positions. METHODS: Respiratory inductive plethysmography was used for quantifying thoraco-abdominal separation and for assessing tidal volumes. 10 subjects were trained to perform widely varying degrees of thoraco-abdominal separation at different tidal volumes. The relationship between the RIIV signal peak-to-peak value (measured at the forearm), and the tidal volume and thoraco abdominal separation was investigated in two body positions with the use of multiple linear regression. RESULTS: Larger tidal volume and more thoracic contribution to respiration were found to increase the RIIV peak-to-peak value (p < 0.0005). In the supine position, the tidal volume influence was stronger than that of thoraco-abdominal separation, and in the sitting position, the opposite was seen. CONCLUSIONS: The effects on the RIIV signal following changes in thoraco-abdominal separation and tidal volume are of the same order of magnitude. In the supine position, the influence of thoracic versus abdominal contribution to the tidal volume is not as significant as in the sitting position. Photoplethysmography is a promising technique for combined monitoring of several respiratory parameters, including tidal volume. In situations where the relative thoracic and abdominal contributions are likely to vary, the tidal volume information becomes less reliable. PMID- 12580234 TI - Hierarchical rule-based monitoring and fuzzy logic control for neuromuscular block. AB - OBJECTIVE: The important task for anaesthetists is to provide an adequate degree of neuromuscular block during surgical operations, so that it should not be difficult to antagonize at the end of surgery. Therefore, this study examined the application of a simple technique (i.e., fuzzy logic) to an almost ideal muscle relaxant (i.e., rocuronium) at general anaesthesia in order to control the system more easily, efficiently, intelligently and safely during an operation. METHODS: The characteristics of neuromuscular blockade induced by rocuronium were studied in 10 ASA I or II adult patients anaesthetized with inhalational (i.e., isoflurane) anaesthesia. A Datex Relaxograph was used to monitor neuromuscular block. And, ulnar nerve was stimulated supramaximally with repeated train-of-four via surface electrodes at the wrist. Initially a notebook personal computer was linked to a Datex Relaxograph to monitor electromyogram (EMG) signals which had been pruned by a three-level hierarchical structure of filters in order to design a controller for administering muscle relaxants. Furthermore, a four-level hierarchical fuzzy logic controller using the fuzzy logic and rule of thumb concept has been incorporated into the system. The Student's test was used to compare the variance between the groups. p < 0.05 was considered significant. RESULTS: The system achieved stable control of muscle relaxation with a mean T1% error of -0.19 (SD 0.66) % accommodating a range in mean infusion rate (MIR) of 0.21-0.49 mg x kg(-1) x h(-1). When these results were compared with our previous ones using the same hierarchical structure applied to mivacurium, less variation in the T1% error (p < 0.05) but the same variation in infusion rate were observed. The controller activity of these two drugs showed no significant difference (p > 0.5). However, the consistent medium coefficient variance (CV) of the MIR of both rocuronium (i.e., 36.13 (SD 9.35) %) and mivacurium (i.e., 34.03 (SD 10.76) %) indicated a good controller activity. CONCLUSIONS: The results showed that a hierarchical rule-based monitoring and fuzzy logic control architecture can provide stable control of neuromuscular block despite the considerable individual variation in neuromuscular block required among patients. Also, there was less variation in T1% error compared with that of previous study on mivacurium. Meanwhile, the consistent medium CV of the MIR of both rocuronium and mivacurium indicated a good controller activity which is able to withstand noise, diathermy effect, artifacts and surgical disturbances. PMID- 12580235 TI - Bispectral index (BIS) and burst suppression: revealing a part of the BIS algorithm. AB - OBJECTIVE: The bispectral index (BIS) is a complex EEG parameter which integrates several disparate descriptors of the EEG into a single variable. One of the subparameters incorporated in the BIS is the suppression ratio, quantifying the percentage of suppression during burst suppression pattern. The exact algorithm used to synthetize the information to the BIS value is unpublished and still unknown. This study provides insight into the integration of the suppression ratio into the BIS algorithm. METHODS: EEG data of 10 healthy volunteers during propofol infusion were analyzed. Propofol concentrations were ramped up to 4 predetermined concentrations (1, 2, 3, 4, 6, 8, 9, or 12 microg/ml) using a computer controlled infusion pump (STANPUMP). EEG recordings were performed with an Aspect A-1000 EEG monitor (Version 3.22). The relationship of the processed EEG variables bispectral index and suppression ratio, calculated by the Aspect A 1000 monitor, was analyzed. RESULTS: Up to 40% suppression ratio the average BIS values remained constant regardless of suppression ratios (r = 0.13). Beyond a suppression ratio of 40%, BIS and suppression ratio were invariably linearly correlated (r = -1). At a suppression ratio > or = 40% the BIS value could be calculated as BIS = 50 - suppression ratio/2. CONCLUSIONS: Suppression ratio values > 40% are linearly correlated with BIS values from 30 to 0. An increasing anesthetic drug effect resulting in an increase of the duration of suppression to a suppression ratio up to 40% is not adequately reflected by the BIS value. PMID- 12580237 TI - Genetic correlations between the maternal genetic effect on chick weight and the direct genetic effects on egg composition traits in a White Leghorn line. AB - Selection can be a useful way to alter yolk proportion and thereby egg dry matter which, owing to its economic importance, is a trait of substantial importance for the egg-processing industry. However, the egg is primarily the chamber of embryonic development. The main purpose of this study was to estimate genetic correlations between the maternal effect on chick weight at hatching and the direct effect on different egg composition traits, in particular, yolk proportion. Additionally, genetic parameters were estimated for egg composition traits. To create a data set suitable for estimation of genetic parameters, a three-round selection experiment was set up. Birds were selected based on their predicted breeding values for the genetic maternal effect on chick weight and the direct genetic effect on yolk proportion according to the theory of elliptical selection. Genetic parameters were estimated using a multiple trait animal model and restricted maximum likelihood. The maternal heritability for chick weight was 0.5, whereas the direct heritability was dose to 0. The genetic correlations between the maternal effect on chick weight and the direct effect on yolk proportion, yolk weight, albumen weight, albumen dry matter concentration, and egg weight were 0.14, 0.76, 0.93, 0.14, and 0.99, respectively. The heritabilities for yolk proportion, yolk weight, albumen weight, albumen dry matter concentration, and egg weight were 0.33, 0.43, 0.57, 0.38, and 0.60, respectively. We conclude that breeding ought to be a useful way to increase egg dry matter with no expected unfavorable correlated effects on chick weight. PMID- 12580236 TI - Neonatal monitoring after maternal fentanyl analgesia in labor. AB - OBJECTIVE: To characterize different methods of monitoring neonatal effects associated with maternal opioid analgesia. Special focus was on the static-charge sensitive bed (SCSB), which could potentially serve as a non-invasive neonatal monitor. METHODS: 12 healthy, term newborns from normal pregnancies were included in this prospective, randomized, controlled study. Maternal labor analgesia was either intravenous fentanyl (n = 5) or paracervical bupivacaine blockade (n = 7). Neonatal recording from delivery to the age of 12 hours included continuous SCSB monitoring with ECG and oximeter for sleep states, respiration, oxygenation, heart rate, and body movements. In addition, umbilical blood pH, Apgar, Amiel Tison's Neurologic and Adaptive Capacity Scoring (NACS), skin cyanosis scoring, blood pressure, rectal and skin temperatures, and skin blood flow measurements were performed. RESULTS: The study was interrupted, because one baby in the fentanyl group had a significant decrease in oxyhemoglobin saturation (SpO2) to 59%. This was considcred to be residual effect of fentanyl and was treated with naloxone. SpO2 was generally lower in the fentanyl group. Epochs with SpO2 < 90% were more frequent in the fentanyl group, especially during active sleep (mean +/ SD 11.9 +/- 10.7% vs. 2.0 +/- 1.7% of epochs, p = 0.034). Mean heart rate values were lower in the fentanyl group (121.1 +/- 6.4 vs. 132.6 +/- 6.8 beats per minute, p = 0.02), and this difference was seen during wake and all sleep states. Maximum heart rate values were lower in the fentanyl group, too. The opiate group had less quiet sleep than controls (9.6 +/- 2.8% vs. 18.3 +/- 8.3%, p = 0.05). NACS after birth was lower in the fentanyl group (median [range] 15 [13-26] vs. 22 [20-25], p = 0.004). CONCLUSIONS: Several differences were seen between the fentanyl and the control group babies. The SCSB method proved sensitive enough to find neonatal effects of maternal analgesia. Together with ECG and SpO2 monitoring, SCSB gives plentiful information on neonatal well-being in a non invasive way. Results of this study emphasize the importance of neonatal monitoring after maternal opiate use in labor. PMID- 12580238 TI - Mapping quantitative trait loci associated with resistance to coccidiosis and growth. AB - To map QTL associated with disease resistance to avian coccidiosis and growth, two commercial broiler lines with different degrees of resistance to the disease were crossed to generate an F1 generation that was intercrossed to produce 314 F2 generation offspring. The F2 offspring were inoculated with sporulated oocysts of Eimeria maxima. Five disease-associated phenotypes were measured after the infection. Intertrait comparisons revealed that oocyst shedding was a good parameter for evaluating disease resistance or susceptibility. One hundred nineteen microsatellite markers, covering 80% of the chicken genome with an average marker interval of 25 cM, were used for genotyping of F1 parents and F2 offspring. Statistical analysis based on the data of four families revealed a locus on chromosome 1 associated with oocyst shedding (logarithm of odds = 3.46). The genetic mechanism of this locus appeared additive. The genomic scan also identified three potential growth QTL on chromosomes 1, 6, and 8. These results provide the foundation for further investigation to validate the QTL. PMID- 12580239 TI - Expression of turkey transcription factors and acyl-coenzyme oxidase in different tissues and genetic populations. AB - Several transcription factors are involved in regulating lipid metabolism in various animal tissues. Peroxisome proliferator-activated receptor (PPAR) gamma and PPARalpha regulate lipogenesis and fatty acid oxidation. Gene fragments for PPARgamma, PPARalpha, and acyl-coenzyme A oxidase (ACO) have been cloned in turkeys, and the sequences of these genes were highly homologous to those of chickens, pigs, and humans. Data showed that turkey PPARgamma and PPARalpha were highly homologous (>97%) with that of the chicken, indicating the genetic relationship between those two species was close. The tissue distribution and genetic effect on mRNA concentrations of PPARgamma, PPARalpha, and ACO in two genetic populations of turkeys [randombred control (RBC2) and growth selected (F line)] was determined. The PPARgamma mRNA was highly expressed in adipose tissue in both populations, but there was no difference between the two populations. The PPARalpha mRNA concentration was high in the liver with less expression in adipose tissue and skeletal muscle. The PPARalpha mRNA concentration was similar between the two genetic populations. The ACO mRNA was expressed in adipose tissue, skeletal muscle, and liver with no difference between the genetic populations. The data suggest that both turkey liver and adipose tissue have considerable capability for fatty acid oxidation and synthesis. Long-term selection for increased 16-wk BW in the F line had no effect on the expression of PPARgamma, PPARalpha, and ACO. PMID- 12580240 TI - Genetic characterization of stress responsiveness in Japanese quail. 1. Analyses of line effects and combining abilities by diallel crosses. AB - The inheritance of stress responsiveness (line effects and combining abilities) was phenotypically studied in progeny from diallel crosses of randombred (RB) quail and quail selected for exaggerated (high stress, HS) or reduced (low stress, LS) plasma corticosterone (CS) response to brief immobilization. The three genotypes were crossed in a 3 x 3 factorial arrangement of treatments that allowed all possible crosses between RB, LS, and HS males with RB, LS, and HS females. The nine crosses produced 479 progeny. At 28 d of age, quail of each cross were stressed by immobilization, and blood was sampled. Plasma CS was used to estimate stress responsiveness in the progeny of each cross. Estimates of the following genetic effects were made: average post-immobilization plasma CS response within each cross, general combining ability (GCA), and specific combining ability (SCA). Differences (P < 0.05) in plasma CS response to immobilization between crosses involving the LS and HS lines (HS crosses > LS crosses) were commonly observed. There were no significant differences in the means of plasma CS responses between males and females within each of the crosses except for the marginal (P = 0.07) sex difference (male > female) found within the LL cross (LS male with LS female). The GCA estimates were -1.23, -2.89, and 5.08 for the RB, LS, and HS quail, respectively. The SCA was not significant for any diallel mating except HH (P < 0.007). Line effects on plasma CS response were different from zero for the LS and HS lines. PMID- 12580241 TI - Genetic characterization of stress responsiveness in Japanese quail. 2. Analyses of maternal effects, additive sex linkage effects, heterosis, and heritability by diallel crosses. AB - Diallel crosses were used to investigate the genetic inheritance of stress responsiveness through examination of population effects in progeny of randombred (RB) quail and quail selected for reduced (low stress, LS) or exaggerated (high stress, HS) plasma corticosterone (CS) response to brief immobilization. The three genotypes were crossed in a 3 x 3 factorial arrangement of treatments that allowed all possible crosses between RB, LS, and HS males with RB, LS, and HS females. The nine crosses produced 479 progeny that were, at 28 d of age, stressed by immobilization, and a sample of blood was collected. The following quantitative genetic parameters were estimated for plasma CS: heritability, heterosis, maternal effects, sex-linkage effects, and heterosis due to the sex chromosomes. Genotypic-phenotypic correlations within RB quail and quail of the two selected lines were also obtained. Maternal effects were different (P < 0.05) from zero for the plasma CS responses of all three genotypes. Additive sex linkage effects on plasma CS responses were variable and of much less importance than maternal effects. The correlations between plasma CS response and genotype were: 0.22 for RB quail and 0.37 and 0.55 for quail of the LS and HS lines, respectively. Heterosis effects were low and most likely due to either an increase in homozygozity of genes on the sex chromosomes or an increase of maternal effects within reciprocal crosses. Heritability, estimated for the post immobilization plasma CS response, was 0.05 for RB quail and 0.14 and 0.30 for quail of the LS and HS lines, respectively. PMID- 12580242 TI - Evaluating the performance and acceptance of teleconference instruction versus traditional teaching methods for undergraduate and graduate students. AB - Institutions are currently seeking alternative ways to deliver a full-line of course materials without acquiring additional staffing. Hence, faculty is charged with creating alternative ways to deliver or offer course content to students. The purpose of this study was to evaluate undergraduate and graduate performance and perception of teleconferencing versus traditional blackboard lectures. In the undergraduate course, we discovered that students performed equally as well on exams and provided favorable reviews of the course; however, the acceptance of this new format is lacking given the enrollment and number of students dropping, e.g., 30 to 40% reduction in the course before semester's end. On the other hand, students taking the graduate course appear to accept the technology well with consistent enrollments and achievement in course content. In summary, using teleconferencing as a way to teach students may be better suited for graduate students when compared to undergraduates. PMID- 12580243 TI - Recursive prediction of broiler growth response to feed intake by using a time variant parameter estimation method. AB - The objective of this study was to explore whether time-variant parameter estimation procedures allow modeling and predicting the dynamic growth response of broiler chickens to feed intake in real time. A recursive linear model was used that estimated the model parameters every 24 h based on a fixed number of actual and past measurements (i.e., time window). Based on 48 datasets, it was concluded that the mean relative prediction error (MRPE) of the recursive linear modeling approach had a minimum for a window size of 5 d. Weight of the birds could be predicted during the growth process 3 to 7 d ahead with a mean relative prediction error of 5% or less. In comparison with the prediction results of three static empirical growth models (one linear and two nonlinear models), the recursive modeling technique had a similar accuracy to the nonlinear empirical models (MRPE of 1.4% to 2.3% vs. 1.1% to 2.8%), but it was less accurate for larger prediction horizons (2 to 7 d). The compact recursive linear model was more accurate than the static linear growth model for prediction horizons of one up to 4 d, depending on the feeding strategy. Since such recursive modeling approach allows the prediction of broiler growth without any prior knowledge of the system and takes into account the time-variant (nonlinear) nature of the growth process based on only a small window of measured information, it is suitable for real-time integration in process management. PMID- 12580244 TI - Improvements in the performance of commercial broilers in the USA: analysis for the years 1997 to 2001. AB - The performance of commercial broilers reported by broiler production units in the USA from 1997 to 2001 has been evaluated. There was a linear decrease in calorie conversion (CC) and number of days to produce a 2.27 kg bird (DAYS) but no change in percentage mortality. By contrast, there was a linear increase in final bird weight (FBW) during this period. There was a linear increase in the percentage of production units rearing birds to more than 2.5 kg and a decrease in those rearing birds to less than 2.0 kg or 2.0 to 2.5 kg. There were no differences in CC or DAYS during the first half of the year (January to June), but these traits showed a significant increase in July followed by a steep decline during August, September, and October. FBW was similar from January to June but showed a significant decrease in July. No consistent variation in monthly mortality was noted. It is concluded that improvements in the productivity of broilers have been achieved from 1997 to 2001 but that more attention should be given to the problem of broiler management during the summer months. PMID- 12580245 TI - Cloning of cDNA encoding the nuclear form of chicken sterol response element binding protein-2 (SREBP-2), chromosomal localization, and tissue expression of chicken SREBP-1 and -2 genes. AB - Sterol regulatory element binding protein-1 and -2 (SREBP-1 and -2) are key transcription factors involved in the biosynthesis of cholesterol and fatty adds. The SREBP have mainly been studied in rodents in which lipogenesis is regulated in both liver and adipose tissue. There is, however, a paucity of information on birds, in which lipogenesis occurs essentially in the liver as in humans. As a prelude to the investigation of the role of SREBP in lipid metabolism regulation in chicken, we sequenced the cDNA, encoding the mature nuclear form of chicken SREBP-2 protein, mapped SREBP-1 and -2 genes and studied their tissue expressions. The predicted chicken SREBP-2 amino acid sequence shows a 77 to 79% identity with human, mouse, and hamster homologues, with a nearly perfect conservation in all the important functional motifs, basic, helix-loop-helix, and leucine zipper (bHLH-Zip) region as well as cleavage sites. As in the human genome, SREBP-1 and SREBP-2 chicken genes are located on two separate chromosomes, respectively microchromosome 14 and macrochromosome 1. Tissue expression data show that SREBP-1 and SREBP-2 are expressed in a wide variety of tissues in chicken. However, unlike SREBP-2, SREBP-1 is expressed preferentially in the liver and uropygial gland, suggesting an important role of SREBP-1 in the regulation of lipogenesis in avian species. PMID- 12580246 TI - Enhanced mucosal immunity against Eimeria acervulina in broilers fed a Lactobacillus-based probiotic. AB - The effect of feeding a Lactobacillus-based probiotic on intestinal intraepithelial lymphocyte (IEL) subpopulations and subsequent protection against coccidiosis was investigated in broiler chickens. Day-old male broilers were fed standard rations without control (CONT) or with a commercial probiotic (PROB) Primalac. Differences in IEL subpopulations were assessed by flow cytometry at 21 d postprobiotic treatment. At 25 d of age, a group of randomly selected birds from each diet was inoculated orally with 10,000 (per bird) sporulated oocysts of Eimeria acervulina and kept on the same diets. Fecal material, sera, and intestinal washes were collected 10 d postchallenge with E. acervulina. Birds on the PROB diet had more IEL expressing the surface markers CD3, CD4, CD8, and alphabetaTCR than those of the CONT diet. The probiotic-fed chickens produced less oocysts (P < 0.0001) compared to the untreated, control group (368 x 10(6) in CONT vs. 89 x 10(6) in PROB). The interferon-gamma levels in both serum and intestinal secretions were not significantly different between the two groups. However, CONT group showed higher antibody levels against a recombinant coccidial antigen in the intestinal secretions than the PROB group. No significant difference was found in serum antibody levels against the same antigen. These results dearly indicate that the probiotic bacteria impacted the local immune response as characterized by altered IEL subpopulations and increased the birds' resistance to E. acervulina as reflected by reduced oocyst shedding. PMID- 12580247 TI - Detection of Salmonella enteritidis-specific immunoglobulin A antibodies in crop samples from chickens infected with Salmonella enteritidis. AB - The crop (ingluvies), an organ for food storage in most avian species when the proventriculus is full, is located at the base of the esophagus. Little is known about any immunological capacity in the crop, and the current study was conducted to determine whether any antibodies to SE could be found in crop flushes taken from White Leghorn hens following infection with this organism. Surprisingly, an exceptionally strong IgA anti-SE response could be detected in the crops of hens 17 d postchallenge, and a comparison at Day 22 of crop vs. intestinal IgA anti-SE responses showed a good correlation between anti-SE antibody levels in the two regions. Histologic examination of crop tissues revealed development of lymphoid aggregates in the crop walls following challenge with SE. These results indicate that the crop may serve a role in immune protection in addition to its capacity as a food storage organ. PMID- 12580248 TI - The effect of different levels of nonphytate phosphorus with and without phytase on the performance of four strains of laying hens. AB - A 4 x 7 factorial experiment was conducted to determine the effect of different nonphytate phosphorus (NPP) regimens with and without phytase on performance of four strains of laying hens. The strains used were Babcock B300, DeKalb Delta White, Hy-Line W36, and ISA-White. The birds of T1 (Treatment 1), control diet, were fed a diet with 0.45% NPP for the entire experiment (20 to 63 wk of age). The birds of T2 were fed a NPP regimen of 0.25% for Phase 1 (20 to 35 wk of age), 0.2% for Phase 2 (36 to 51 wk of age), and 0.15% for Phase 3 (52 to 63 wk of age). The birds of T3 and T4 were fed a NPP regimen similar to T2 plus 150 or 300 units phytase/kg diet, respectively. The birds of T5 were fed a NPP regimen of 0.2, 0.1, and 0.1% for Phases 1, 2, and 3, respectively. The birds on T6 and T7 were fed a NPP regimen similar to T5, with 150 and 300 units phytase/kg diet, respectively. The criteria for evaluating the effect of dietary treatments were production performance, shell quality, bone ash, and total P excretion. Production traits were drastically reduced and mortality was drastically increased when the birds of T5 were fed a diet with 0.1% NPP during Phase 2 (36 to 51 wk of age). Increasing the NPP level of this treatment from 0.1 to 0.45% during Phase 3 restored the production traits almost to the control level during this phase of the experiment. The NPP regimen of 0.2-0.1-0.1% plus either level of phytase (T6 and T7) restored most of the traits for the entire experiment to the control level (T1), except egg production (EP) and the nonprolapse mortality. Bone ash also remained significantly lower than the control group with this NPP regimen plus the higher level of phytase (T7). Production traits were reduced, and nonprolapse mortality was increased due to the use of a NPP regimen of 0.25 0.2-0.15% without phytase (T2) for the entire experiment. The NPP regimen of 0.25 0.2-0.15% plus the lower level of phytase (T3) restored all the traits except EP to the control level for the entire experiment. The only treatment that maintained performance of all the strains comparable to their controls for the entire experiment was a NPP regimen of 0.25-0.2-0.15% plus the higher level of phytase (T4). Total P excretion of birds fed this NPP regimen was reduced by 55.6% as compared to the control group. The data generally indicated that the higher level of phytase was more effective than the lower level in restoring the performance of birds fed the low-P diets to the control level. Numerous interactions existed among strain x diet for various traits throughout the experiment, indicating the NPP requirement for maintaining production performance may be different among strains. PMID- 12580249 TI - Effects of phytase supplementation on the performance, egg quality, and phosphorous excretion of laying hens fed different levels of dietary calcium and nonphytate phosphorous. AB - An experiment employing a factorial arrangement of two levels (3.0 and 4.0%) of Ca, two levels (0.15 and 0.25%) of nonphytate phosphorus (NPP), and two levels (0 and 300 U/kg diet) of microbial phytase was carried out with 960 ISA-brown layers from 21 to 41 wk of age. There was a significant interaction between NPP level and phytase for egg production. High NPP level and phytase supplementation increased egg production only in the second 10-wk period (31 to 41 wk). High NPP and low Ca increased feed intake, and a significant interaction between levels of NPP and Ca was observed in the first 10 wk. High NPP improved feed efficiency only in the second 10-wk period. Low NPP improved egg specific gravity and eggshell thickness but decreased Haugh units in the first 10-wk period; high NPP decreased the percentage of broken and soft-shell eggs in the second period. Low Ca decreased egg specific gravity, eggshell strength, and eggshell thickness in both periods and increased Haugh units in the second 10-wk period. Phytase supplementation decreased the percentage of broken and soft-shell eggs. High NPP increased fiber availability but decreased Ca availability. High Ca decreased Ca availability, whereas phytase increased availability of dry matter, fiber, and P. High NPP increased retention of P and Fe but also increased excretion of P. High Ca decreased retention of Zn and Fe. Phytase supplementation increased P retention, resulting in decrease of P excretion. In conclusion, supplementation of microbial phytase at a level of 300 U per kg diet of laying hens can improve egg production, decrease broken and soft egg production rate, and P excretion. The effects of phytase supplementation are significantly modified by the level of Ca and NPP. PMID- 12580250 TI - Interactions of various supplies of isoleucine, valine, leucine and tryptophan on the performance of laying hens. AB - The present study was undertaken to investigate the interactions among the supplies of isoleucine, leucine, valine, and tryptophan in laying hens. A three factor trial was conducted with laying hens in which the dietary concentrations of isoleucine (5.7, 8.0, and 11.5 g/kg), valine and leucine (6.3 and 7.2 g/kg and 10.1 and 11.5 g/kg, respectively), and tryptophan (1.5 and 2.4 g/kg) were varied. At the lowest concentration of valine + leucine, an increase in dietary isoleucine concentration led to a dose-dependent reduction in feed consumption, daily egg mass, and body weight gain and an increase of the isoleucine concentration in plasma. At a high dietary concentration of valine + leucine, excess dietary isoleucine concentration caused only a weak depression of performance parameters; the isoleucine concentration in plasma was independent of the dietary isoleucine concentration. Increasing the dietary tryptophan concentration did not influence the effect of an excessive dietary isoleucine concentration on performance parameters. Increasing the tryptophan concentration from 1.5 to 2.4 g/kg diet did, however, lead to a significant increase in feed consumption, irrespective of the supply of isoleucine, valine, and leucine. In conclusion, our study demonstrates that the supply of valine + leucine influenced the effects of excess dietary isoleucine in laying hens, whereas the supply with tryptophan did not. PMID- 12580251 TI - Hypoglycemia and reduced feed intake in broiler chickens treated with metformin. AB - The bi-guanide metformin is used to treat noninsulin dependent diabetes in obese patients. In addition to having antihyperglycemic effects, metformin is also anorectic and reduces BW. These studies were performed to determine if metformin possesses similar properties in chickens. Metformin-HCl was administered to 14 day-old broiler chickens at either 300 or 600 mg/kg per day in the drinking water for 10 d while monitoring BW and feed intake. No changes in water intake were observed, while feed intake and daily gains were only significantly reduced by the 600 mg/kg dose. After oral administration of a single dose of 300 mg/kg metformin-HCl, feed intake was significantly reduced by 4 h and remained suppressed for greater than 24 h relative to controls. Plasma hormones and metabolites (glucose, lactate, insulin, glucagon, uric acid, nonesterified fatty acid, and triglycerides) were monitored at 1, 2, 3, 6, and 24 h posttreatment. Significant and acute decreases in blood glucose, insulin, and triglycerides were observed at 3 h posttreatment as compared to controls. Opposing acute increases in glucagon and NEFA levels were also observed at 3 h followed by an increase in uric add 6 h posttreatment. These observations suggest that metformin induces metabolic changes in birds, similar to that observed in mammals and may act in a common manner. Metformin-HCl may be useful in glucose metabolism studies by inducing hypoglycemia, a condition rarely observed in birds. PMID- 12580252 TI - Dietary conjugated linoleic acid alters hepatic lipid content and fatty acid composition in broiler chickens. AB - We examined the effect of dietary conjugated linoleic acid (CLA) on growth performance and liver composition in broiler chickens. Day-old male broiler chicks were assigned to receive a diet supplemented with corn oil (5%; n = 48) or CLA (5%; n = 48) for 21 d. Broilers fed CLA weighed less and grew at slower rates than broilers fed corn oil. Feed intake and feed conversion were higher for the corn-oil group than for the CLA dietary group. Hepatic lipid and triacylglycerol concentrations were significantly reduced by dietary CLA. The proportions of saturated fatty acids (SFA) in liver lipids increased, whereas those of monounsaturated fatty acids (MUFA) decreased with CLA supplementation. Although the total concentration of polyunsaturated fatty acids (PUFA) did not change with dietary treatment, the concentration of linoleic acid as a percentage of total methylated fatty acids decreased, and that of linolenic add increased in broilers fed CLA. The concentration of CLA isomers in liver lipids increased substantially with CLA feeding. The relative proportion of the c9,t11 CLA isomer in hepatic lipids was much higher than that of the t10,c12 or t9,t11 CLA isomers. These studies provide evidence that feeding CLA to broilers results in substantial reduction in liver fat accumulation and promotes CLA incorporation into hepatic lipid pools. PMID- 12580253 TI - Fat and carbohydrate use in posthatch chicks. AB - Metabolism of carbohydrate and fat was examined in the hatching chick by introduction of 14C labeled oleic acid, triolein, or glucose to the gastrointestinal tract or to the yolk sac. Label in plasma and exhaled air samples were determined over a 12-h period. The activity-time curves could be fitted to double exponential equations, and rate constants for appearance and disappearance of label were calculated. Following injection to the yolk, oleic acid and triolein label showed rapid appearance in the circulation and rapid utilization. The rate of appearance of oleic acid and triolein in the circulation from the gastrointestinal tract was high at hatch and did not change with age. However, the rate constant for disappearance from the plasma increased with age and after 6 d posthatch disappearance curves fluctuated with little decline over 12 h. Following the introduction of labeled lipids more label was found in acylglycerides with time after injection but age had no effect. Lipids introduced via the yolk sac were present in higher proportions in fractions corresponding to low-density lipoprotein (LDL) than lipids introduced via the gastrointestinal tract. Glucose appearance in the circulation from the gastrointestinal tract was low at hatch and increased more than two-fold by 3 d posthatch. The rate constant for disappearance of glucose from the circulation did not change with age. It appears that lipoproteins synthesized maternally or in yolk transport lipids at hatch, but their concentration decreases posthatch, and this could reduce utilization of circulating lipids. Uptake of glucose increases after hatch, thus, allowing glucose to become a major source of energy as the yolk lipids are depleted. PMID- 12580254 TI - Free diet selection by broilers as influenced by dietary macronutrient ratio and corticosterone supplementation. 1. Diet selection, organ weights, and plasma metabolites. AB - Male broiler chickens (aged 21 d) were allowed to chose freely for 14 d between three diets in which only one specific macronutrient (protein, lipid, or carbohydrate) was isocalorically substituted for one other macronutrient, but otherwise (nearly) isocaloric and composed of the same ingredients. The three diets were low protein (LowCP; 15.81% CP; 6.56% lipid; 50.78% carbohydrate), low lipid (LowL; 19.63% CP; 3.01% lipid; 51.12% carbohydrate), and low carbohydrate (LowCHO; 19.50% CP; 7.72% lipid; 44.00% carbohydrate). The chickens either received 0, 30, or 45 mg of corticosterone (CORT) per kg diet. As a percentage of their total intake, unsupplemented chickens consumed 24.0, 71.4, and 4.6% of the LowCP, LowL, and LowCHO diets, respectively, giving a total CP, L, and CHO intake of 282, 61, and 765 g, respectively. The addition of CORT significantly changed the diet selection, as compared to the unsupplemented chickens, CORT chickens consumed a greater percentage from the LowCP (35%), less from the LowL (55%), and again more from the Low-CHO (10%) diet. On the other hand, total feed consumption, macronutrient, and ME intake were not altered significantly by CORT supplementation, probably because of the close similarity of the diets. Corticosterone-supplemented chickens manifested hyperglycemia, hyperlipidemia, and uric acidemia suggesting insulin resistance, increased lipogenesis and protein catabolism, respectively. The elevated plasma creatine kinase (CK) activities of CORT chickens are also suggestive for decreased muscle cell membrane stability. Furthermore, CORT chickens were characterized by increased proportional weights of liver, abdominal fat pad, proventriculus, and gizzard, whereas an involution of spleen and bursa was observed. In conclusion, the present results suggest that high circulating levels of CORT as in the case of stress results in metabolic alterations, which in turn, affects diet preference as a compensatory mechanism to adapt energy and nutrient metabolism. PMID- 12580255 TI - Influence of enzymes on performance and digestive parameters of broilers fed rye based diets. AB - An experiment was conducted to study the influence of enzyme supplementation (ES) to rye-based diets on rate of food passage through the digestive tract, viscosity of jejunum content, volatile fatty acid concentration in ceca, and performance of broilers. There were seven treatments; six diets arranged factorially with three varieties of rye (Petkus, Prima, and Saratov V) and two levels of ES (0 or 500 ppm of an enzyme complex containing 858 IU of beta-glucanase and 864 IU of xylanase/g) and an additional control diet based on corn. Each treatment was replicated seven times (12 chicks caged together), and the trial lasted 25 d. Rye feeding increased intestinal viscosity and impaired bird performance at 25 d (P < 0.001). Among rye diets the greatest feed intake and weight gain were obtained with Petkus variety, which also produced the lowest intestinal viscosity. Enzyme addition reduced the time needed to recover 1% (0.78 vs. 0.98 h; P < 0.05) and 50% (4.2 vs. 6.5 h; P < 0.01) of the marker in feces and reduced the mean retention time of marker in the gastrointestinal tract (17.1 vs. 18.8 h; P < 0.05). Also, ES reduced intestinal viscosity (P < 0.001) and improved feed intake, daily gain, and feed conversion of birds from 4 to 25 d (P < 0.01) but did not modify volatile fatty acid concentration in ceca. We concluded that ES added to rye diets decreased intestinal viscosity and accelerated digestive transit, improving productive performance of broilers. PMID- 12580256 TI - Nutritional evaluation of dried tomato seeds. AB - Two samples of tomato seeds, a by-product of the tomato canning industry were evaluated to determine proximate analysis, amino acid content, and digestibility, TMEn, and protein efficiency ratio. Tomato seeds were also used to replace corn and soybean meal (SBM) in a chick diet on an equal true amino acid digestibility and TMEn basis. Tomato seeds were found to contain 8.5% moisture, 25% CP, 20.0% fat, 3.1% ash, 35.1% total dietary fiber, 0.12% Ca, 0.58% P, and 3,204 kcal/kg of TMEn. The total amounts of methionine, cystine, and lysine in the tomato seeds were 0.39, 0.40, and 1.34%, respectively, and their true digestibility coefficients, determined in cecectomized roosters, were 75, 70, and 54%, respectively. The protein efficiency ratio (weight gain per unit of protein intake) value when fed to chicks at 9% CP was 2.5 compared to 3.6 for SBM (P < or = 0.05). When corn-SBM diets were formulated on an equal true amino acid digestibility and TMEn basis, up to 15% tomato seeds could replace corn and SBM without any adverse affects on chick weight gain, feed intake, or gain:feed ratio from 8 to 21 d posthatch. Tomato seeds at any level in the diet did not significantly affect skin pigmentation. Although the protein quality of tomato seeds may not be as high as SBM, tomato seeds do contain substantial amounts of digestible amino acids and TMEn. When formulating diets on a true digestible amino acid and TMEn basis, tomato seeds can be supplemented into chick rations at up to 15% without any adverse affects on growth performance. PMID- 12580257 TI - Performance of two strains of laying hens fed ground and whole barley with and without access to insoluble grit. AB - The live performance from 19 to 43 wk of age of two strains of commercial White Leghorn hens fed two levels of whole barley (0 or 60%) and insoluble grit (0 or 4 g/bird per wk) was compared. The 0 and 60% whole barley diets differed only in feed form and were formulated to the same nutrient specifications. No dilution of nutrients or ingredients occurred. The 0% whole barley diet was fed in mash form. The 60% whole barley diet was fed as whole grain and mash concentrate blended into a complete diet and fed in the same feed trough. Feeding whole barley reduced egg production, feed efficiency, and egg specific gravity and increased feed intake, egg weight, and body weight gain. Access to insoluble grit had no effect on any of the production variables measured. The two strains of hens responded similarly to whole barley but differed in feed intake, feed efficiency, egg weight, egg specific gravity, and body weight gain. Feeding whole barley combined with a mash concentrate depressed hen performance compared to birds fed a similar diet in mash form. Strain of hen and access to insoluble grit did not alter the response to feeding whole barley. PMID- 12580258 TI - Immune function in turkey breeder hens during the short day prelighting period and renewal of photosensitivity for egg production. AB - Photorefractoriness (PR) in the turkey breeder hen is characterized by a lack of responsiveness to photoperiods that previously induced or maintained egg production. The consequence of PR is spontaneous regression of ovarian function and cessation of lay. Photosensitivity (PS) may be regained by giving at least 8 wk of short photoperiod (8L:16D) (light restriction). Following the transition from PR to PS, the birds may be photostimulated with long photoperiods, which allows for the recrudescence of ovarian function and normal egg production. Although the return of reproductive viability is the parameter for determining the successful recycle of ovarian function, there are no known reports of the physiological costs of this transition on immune function in the turkey breeder hen. We conducted an experiment to determine the immune responsiveness at various stages of recycle in the turkey breeder hen. Fifty photorefractory birds were selected and distributed equally among five treatment groups (time points). All birds were given an 8-wk period of light restriction (8L:16D) followed by a 12-wk period of photostimulation (16L:8D). The cellular (cutaneous basophil hypersensitivity CBH) and humoral (antibody titer) immune responses were determined in each treatment group (sequential time points): prelight restriction, 2-wk light restriction, 7-wk light restriction, 2-wk photostimulation, and 12-wk photostimulation. After 2-wk light restriction, there was a reduction in the cellular (64.1%) and humoral (59.5%) immune responses from that of the PR hens at the start. After 7-wk light restriction, the humoral responses increased (33.5%) as compared to the 2-wk light restriction time point Upon photostimulation, both the cellular (23.3%) and humoral (52.4%) immune responses were reduced at 2 wk of photostimulation as compared to the prior 7-wk light restriction time point. Finally, there was a rise in cellular (45.7%) and humoral (72.3%) immune responses after 12 wk of photostimulation as compared to the prior 2-wk photostimulation time point. We concluded that recycling of PR turkey hens was associated with altered cellular and humoral immune responses characterized by initial decline then recovery in both the light restriction and the postphotostimulation periods. PMID- 12580259 TI - Tissue distribution of prolactin receptor mRNA during late stage embryogenesis of the chick. AB - Serum prolactin increases during late embryogenesis. In order to elucidate the function of prolactin at this period, tissue distribution of prolactin receptor mRNA was examined by RNase protection assay. The mRNA was detected strongly in the kidney, intestine, and allantoic membrane; weakly detected in the brain; but not detected in the liver. The expression levels of the prolactin receptor mRNA in the kidney, intestine, and allantoic membrane were retained at constant levels during later stages of embryogenesis (Days 17 and 19) and posthatch periods (2 and 28 d after hatching). These results suggest that prolactin is mainly involved in the osmoregulation during the later stage of embryogenesis and that the expression of prolactin receptor mRNA in the kidney, intestine, and allantoic membrane is regulated by a serum prolactin-independent manner. PMID- 12580260 TI - The effect of electrolyzed oxidative water applied using electrostatic spraying on pathogenic and indicator bacteria on the surface of eggs. AB - Research was conducted to compare the effectiveness of electrolyzed oxidative (EO) water applied using an electrostatic spraying system (ESS) for killing populations of bacteria that are of concern to the poultry industry. Populations of pathogenic bacteria (Salmonella typhimurium, Staphylococcus aureus, and Listeria monocytogenes), and the indicator bacterium Escherichia coli were applied to eggs and allowed to attach for 1 h. EO water completely eliminated all Salmonella typhimurium on 3, 7, 1, and 8 out of 15 eggs in Repetitions (Rep) 1, 2, 3, and 4, respectively, even when very high inoculations were used. EO water completely eliminated all Staphylococcus aureus on 12, 11, 12, and 11 out of 15 eggs in Rep 1, 2, 3, and 4, respectively. EO water completely eliminated all Listeria monocytogenes on 8, 13, 12, and 14 out of 15 eggs in Reps 1, 2, 3, and 4, respectively. EO water completely eliminated all Escherichia coli on 9, 11, 15, and 11 out of 15 eggs in Reps 1, 2, 3, and 4, respectively. Even when very high concentrations of bacteria were inoculated onto eggs (many times higher than would be encountered in industrial situations), EO water was found to be effective when used in conjunction with electrostatic spraying for eliminating pathogenic and indicator populations of bacteria from hatching eggs. PMID- 12580261 TI - Effects of stunning and decapitation on broiler activity during bleeding, blood loss, carcass, and breast meat quality. AB - Four experiments were conducted to determine the effects of electrical stunning and decapitation on bird activity as well as carcass and meat quality. In Experiment 1, broilers were subjected to one of four stunning and killing methods: no stun and neck cut, stun and neck cut, no stun and decapitation, and stun and decapitation. Birds were scored for severity of physical activity on a scale of 1 to 4 with 1 being no activity and 4 being severe wing flapping and muscular contractions. Carcasses were also scored for red wing tips and broken bones. In Experiments 2 to 4, all birds were stunned prior to neck cut or decapitation. Carcasses were scored as described in Experiment 1 as well as measurements of blood loss, feather removal, and breast meat pH, color, cook loss, and tenderness. Based on carcass activity in Experiment 1, decapitation following stunning was similar to a conventional stun and unilateral neck cut, except there was almost no late activity (after 60 s) observed in the decapitated birds. Decapitation following stunning did not result in any consistent carcass quality defects compared to conventional killing in the four experiments. No differences were found in 24-h lightness values, yellowness, cook yield, tenderness, and ultimate pH between conventionally killed and decapitated birds. Blood loss and breast meat redness were inconsistent. These results indicate that high frequency stunning and decapitation may be an acceptable alternative to conventional slaughter based on carcass and meat quality and by ensuring an irreversible loss of consciousness. PMID- 12580262 TI - Effect of broiler age, feed withdrawal, and transportation on levels of coliforms, Campylobacter, Escherichia coli and Salmonella on carcasses before and after immersion chilling. AB - A study was conducted to determine effects of bird age at slaughter, feed withdrawal, and transportation on levels of coliforms, Campylobacter, Escherichia coli, and Salmonella on carcasses before and after immersion chilling. Broilers were processed at 42, 49, and 56 d of age after a 12-h feed withdrawal period or a 0-h feed withdrawal period (full fed). At each age, broilers were processed from two commercial farms previously identified as Campylobacter positive. One week before slaughter, broilers were gavaged with nalidixic acid-resistant Salmonella. During bleeding, cotton plugs were inserted into the cloaca of each carcass. Whole-carcass rinses (WCR) were performed before and after immersion chilling with 20 ppm sodium hypochlorite, and rinses were analyzed for coliforms, Campylobacter, E. coli and Salmonella. Log10 counts for coliforms, Campylobacter, and E. coli were (P < 0.05) affected by bird age at slaughter. Feed withdrawal (FW) affected only Campylobacter on carcasses of older broilers (56 d of age). Chilling with sodium hypochlorite resulted in log10 reductions of 1.2, 1.3, 1.4, and 0.5 for coliforms, Campylobacter, E. coli, and Salmonella, respectively. Under the conditions of this experiment, it appears that contamination on the exterior of birds entering the processing facility is critical to carcass bacterial counts. Moreover, carcass bacterial counts did not vary when microbial counts of broilers were comparable. FW may increase prechill carcass counts for E. coli and Campylobacter, but it appears to have no effect on postchill carcass counts when sodium hypochlorite is used in the chilling operation. PMID- 12580263 TI - The efficacy of conventional rhegmatogenous retinal detachment surgery in the pediatric population. PMID- 12580264 TI - Retinoblastoma. PMID- 12580265 TI - Ten-year surgical experience with childhood glaucoma. PMID- 12580266 TI - Magnetic resonance imaging in Duane's retraction syndrome. AB - PURPOSE: The aim of this study was to visualize the subarachnoid portion of the nervus abducens by magnetic resonance imaging and to analyze whether aplasia of the nervus abducens is an etiologic factor in Duane's retraction syndrome. METHODS: We performed thin-sectioned magnetic resonance imaging across the brainstem level in 8 cases (11 eyes) that were clinically diagnosed as Duane's retraction syndrome. The same test was applied to 8 healthy control subjects to verify the accuracy of this technique. RESULTS: The nervus abducens on the affected side could not be observed in 6 (54.5%) of 11 eyes (8 cases) that were clinically diagnosed as having Duane's retraction syndrome. The nervus abducens was observed in 15 (94%) of 16 eyes that were screened as the control group. CONCLUSIONS: The results showed that aplasia of the nervus abducens, although an important etiologic factor, is not the only factor responsible for the diagnosis of Duane's retraction syndrome. PMID- 12580267 TI - Comparison of anti-emetic effects of ondansetron and low-dose droperidol in pediatric strabismus surgery. AB - BACKGROUND AND OBJECTIVE: Strabismus surgery is associated with a high incidence of postoperative nausea and vomiting. A dose of 150 microg/kg(-1) of ondansetron has been found to be effective in reducing nausea and vomiting in pediatric patients. However, droperidol (20 to 75 microg/kg(-1)) has shown variable success with various side effects. This randomized, double-blind, placebo-controlled clinical trial was conducted to evaluate the efficacy and safety of 25 microg/kg( 1) of droperidol and compare it with that of 150 microg/kg(-1) of ondansetron in pediatric strabismus surgery. PATIENTS AND METHODS: After institutional approval and parental informed consent, 120 children aged 1 to 15 years were prospectively randomized into one of three groups. Group 1 received normal 'saline (placebo), group 2 received 25 microg/kg(-1) of droperidol, and group 3 received 150 microg/kg(-1) of ondansetron after induction of anesthesia. Children were assessed postoperatively for the incidence and severity of vomiting, recovery from anesthesia, and analgesic requirement for 24 hours. RESULTS: There was a significant (P < .05) reduction in the incidence of postoperative nausea and vomiting in the ondansetron (37.5%) and droperidol (30%) groups compared with the placebo (62.5%) group. The vomiting was also significantly less severe in these groups. CONCLUSION: Doses of 25 microg/kg(-1) of droperidol and 150 microg/kg(-1) of ondansetron administered at induction of anesthesia are equally effective in reducing the incidence and severity of postoperative nausea and vomiting in children undergoing strabismus surgery. PMID- 12580268 TI - Subtenon injection of botulinum toxin for treatment of traumatic sixth nerve palsy. AB - PURPOSE: Subtenon injection of botulinum toxin may produce results similar to intramuscular injection of the medial rectus muscle for the treatment of acute traumatic sixth nerve palsy. This study was designed to evaluate the clinical efficacy of subtenon injection and to compare our results with those in previously published reports. METHODS: During 3 years at a single institution, 13 patients with traumatic sixth nerve palsy of less than 6 months' duration were treated with subtenon injection of botulinum toxin. The deviation angles before and after injection were recorded. A distance esotropia of less than 10 prism diopters (PD) in the primary position or absence of diplopia at 3 months was defined as recovery. RESULTS: Of the 13 patients treated, 11 (84.5%) had unilateral palsy and 2 (15.4%) had bilateral palsy. The average pre-injection deviation was 39.5 PD of esotropia, and the average post-injection deviation was 17.0 PD. Seven patients experienced recovery and regained binocular single vision; the overall recovery rate was 53.8% (unilateral, 63.6%; bilateral, 0%). Six patients did not recover and subsequently underwent strabismus surgery. CONCLUSION: Patients with traumatic sixth nerve palsy treated with subtenon injection of botulinum toxin showed higher recovery rates than did most patients treated with conservative measures in published reports. The result of subtenon injection of botulinum toxin without electromyography (EMG) guidance was comparable to that obtained using EMG-guided intramuscular injection of botulinum toxin. Patients with unilateral palsy demonstrated a better recovery rate than did patients with bilateral palsy. PMID- 12580269 TI - Surgical repair of a blocked pupil in Rieger's anomaly. PMID- 12580271 TI - Kinematic magnetic resonance imaging in Y pattern exodeviations. PMID- 12580270 TI - Can the visual acuity of infants be predicted from a measurement of contrast sensitivity? PMID- 12580272 TI - Bilateral eccentric fixation with exotropia in a patient with bilateral healed central chorioretinitis. PMID- 12580273 TI - Suprasellar dermoid cyst presenting as acquired exotropia. PMID- 12580274 TI - An ophthalmic screening protocol for nail-patella syndrome. PMID- 12580275 TI - Central retinal artery occlusion associated with leukemic optic neuropathy. PMID- 12580276 TI - Optic nerve glioma in a patient with von Hippel-Lindau syndrome. PMID- 12580277 TI - Agrosuppression: a bioassay for the hypersensitive response suited to high throughput screening. AB - We describe a novel method, agrosuppression, that addresses the need for an assay of the hypersensitive response (HR) in intact plants that is rapid and adapted to high-throughput functional screening of plant and pathogen genes. The agrosuppression assay is based on inoculation of intact plants with a mixture of Agrobacterium tumefaciens strains carrying (i) a binary plasmid with one or more candidate HR-inducing genes and (ii) a tumor-inducing (oncogenic) T-DNA. In the absence of HR induction, tumor formation is initiated, resulting in a typical crown gall phenotype. However, upon induction of the HR, tumor formation by the oncogenic T-DNA is suppressed, resulting in a phenotype that can be readily scored. We tested and optimized agrosuppression in Nicotiana benthamiana using the inf1 elicitin gene from the oomycete pathogen Phytophthora infestans, which specifically induces the HR in Nicotiana spp., and the gene-for-gene pair Avr9/Cf 9 from the fungal pathogen Cladosporium fulvum and Lycopersicon pimpinellifolium (currant tomato), respectively. Agrosuppression protocols that can be rapidly performed using simple mechanical wounding of petioles of intact N. benthamiana plants were developed and appeared particularly adapted to intensive high throughput screening. This assay promises to greatly facilitate the cloning of novel plant R genes and pathogen Avr genes and to accelerate functional analyses and structure-function studies of these genes. PMID- 12580278 TI - Candidate defense genes from rice, barley, and maize and their association with qualitative and quantitative resistance in rice. AB - Candidate genes involved in both recognition (resistance gene analogs [RGAs]) and general plant defense (putative defense response [DR]) were used as molecular markers to test for association with resistance in rice to blast, bacterial blight (BB), sheath blight, and brown plant-hopper (BPH). The 118 marker loci were either polymerase chain reaction-based RGA markers or restriction fragment length polymorphism (RFLP) markers that included RGAs or putative DR genes from rice, barley, and maize. The markers were placed on an existing RFLP map generated from a mapping population of 116 doubled haploid (DH) lines derived from a cross between an improved indica rice cultivar, IR64, and a traditional japonica cultivar, Azucena. Most of the RGAs and DR genes detected a single locus with variable copy number and mapped on different chromosomes. Clusters of RGAs were observed, most notably on chromosome 11 where many known blast and BB resistance genes and quantitative trait loci (QTL) for blast, BB, sheath blight, and BPH were located. Major resistance genes and QTL for blast and BB resistance located on different chromosomes were associated with several candidate genes. Six putative QTL for BB were located on chromosomes 2, 3, 5, 7, and 8 and nine QTL for BPH resistance were located to chromosomes 3, 4, 6, 11, and 12. The alleles of QTL for BPH resistance were mostly from IR64 and each explained between 11.3 and 20.6% of the phenotypic variance. The alleles for BB resistance were only from the Azucena parent and each explained at least 8.4% of the variation. Several candidate RGA and DR gene markers were associated with QTL from the pathogens and pest. Several RGAs were mapped to BB QTL. Dihydrofolate reductase thymidylate synthase co-localized with two BPH QTL associated with plant response to feeding and also to blast QTL. Blast QTL also were associated with aldose reductase, oxalate oxidase, JAMyb (a jasmonic acid-induced Myb transcription factor), and peroxidase markers. The frame map provides reference points to select candidate genes for cosegregation analysis using other mapping populations, isogenic lines, and mutants. PMID- 12580279 TI - Localization of a potyvirus and the viral genome-linked protein in wild potato leaves at an early stage of systemic infection. AB - The upper noninoculated 'sink' leaves of the wild potato species, Solanum commersonii, were studied for distribution of Potato virus A (PVA) at an early stage of systemic infection. Viral RNA was detected by in situ hybridization, and five viral proteins were localized using immunohistochemical staining in leaf sections. Initial systemic infection foci were found at the vicinity of major and minor veins. In these infection foci, the viral coat protein, cylindrical inclusion protein, and helper component-proteinase colocalized with viral RNA in parenchyma and mesophyll cells, but none of these were detected in companion cells (CC). In contrast, VPg, which is the N-proximal half of the NIa protein (separated from the C-terminal proteinase domain, NIapro, by an autocatalytic cleavage) and acts as a viral genome-linked protein, was detected in CC in the infection foci, but only at an early stage of virus unloading. Outside the infection foci, conspicuous signals for VPg were readily and exclusively detected in CC of many veins in all vein classes in the absence of signals for NIapro, other viral proteins, and viral RNA. Taken together, our data indicate that both major and minor veins may unload PVA in the sink leaves of potato. The data suggest that VPg is translocated from inoculated source leaves to the sink leaves, where it accumulates in CC at an early stage of systemic infection. These findings suggest that VPg may be a 'phloem protein' that specifically acts in CC in the sink leaves to facilitate virus unloading. PMID- 12580280 TI - Differential induction of symptoms in Arabidopsis by P6 of Cauliflower mosaic virus. AB - The gene VI protein (P6) of Cauliflower mosaic virus (CaMV) functions as a virulence factor in crucifers by eliciting chlorotic symptoms in infected plants. The ability to induce chlorosis has been associated previously with P6 through gene-swapping experiments between strains and through the development of transgenic plants that express P6. The primary role that has been identified for P6 in the CaMV infection cycle is to modify the host translation machinery to facilitate the translation of the polycistronic CaMV 35S RNA. This function for P6 has been designated as the translational transactivator (TAV) function. In the present study, we have characterized an unusual variant of P6, derived from CaMV strain D4, that does not induce chlorosis upon transformation into Arabidopsis thaliana. The level of D4 P6 produced in transgenic Arabidopsis line D4-2 was comparable to the amount found in transgenic plants homozygous for W260 and CM1841 P6, two versions of P6 that induce strong chlorotic symptoms and stunting in Arabidopsis. A complementation assay proved that P6 expressed in the D4-2 line was functional, as it could support the systemic infection of a CM1841 mutant that contained a lethal frame-shift mutation within gene VI. This complementation assay allowed us to separately assess the contribution of CM1841 gene VI to symptom development versus the contribution of other CM1841 genes. Furthermore, a previous study had shown that the TAV activity of D4 P6 was comparable to that of W260 P6. That comparative analysis of TAV function, coupled with the characterization of the D4-2 transgenic line in the present paper, indicates that the TAV function of P6 may play only a minor role in the development of chlorotic symptoms. PMID- 12580281 TI - A pseudomonas syringae pv. tomato DC3000 Hrp (Type III secretion) deletion mutant expressing the Hrp system of bean pathogen P. syringae pv. syringae 61 retains normal host specificity for tomato. AB - The plant pathogenic species Pseudomonas syringae is divided into numerous pathovars based on host specificity. For example, P. syringae pv. tomato DC3000 is pathogenic on tomato and Arabidopsis, whereas P. syringae pv. syringae 61 is pathogenic on bean. The ability of P. syringae strains to elicit the hypersensitive response (HR) in non-hosts or be pathogenic (or parasitic) in hosts is dependent on the Hrp (type III secretion) system and effector proteins this system is thought to inject into plant cells. To test the role of the Hrp system in determining host range, the hrp/hrc gene cluster (hrpK through hrpR) was deleted from DC3000 and complemented in trans with the orthologous cluster from strain 61. Mutant CUCPB5114 expressing the bean pathogen Hrp system on plasmid pCPP2071 retained the ability of wild-type DC3000 to elicit the HR in bean, to grow and cause bacterial speck in tomato, and to elicit a cultivar specific (gene-for-gene) HR in tomato plants carrying the Pto resistance gene. However, the symptoms produced in compatible tomato plants involved markedly reduced chlorosis, and CUCPB5114(pCPP2071) did not grow or produce symptoms in Arabidopsis Col-0 although it was weakly virulent in NahG Arabidopsis. A hypersensitive-like collapse was produced by CUCPB5114(pCPP2071) in Arabidopsis Col-0 at 1 x 10(7) CFU/ml, but only if the bacteria also expressed AvrB, which is recognized by the RPM1 resistance gene in Col-0 and confers incompatibility. These observations support the concept that the P. syringae effector proteins, rather than secretion system components, are the primary determinants of host range at both the species and cultivar levels of host specificity. PMID- 12580282 TI - Infection-blocking genes of a symbiotic Rhizobium leguminosarum strain that are involved in temperature-dependent protein secretion. AB - Rhizobium leguminosarum strain RBL5523 is able to form nodules on pea, but these nodules are ineffective for nitrogen fixation. The impairment in nitrogen fixation appears to be caused by a defective infection of the host plant and is host specific for pea. A Tn5 mutant of this strain, RBL5787, is able to form effective nodules on pea. We have sequenced a 33-kb region around the phage transductable Tn5 insertion. The Tn5 insertion was localized to the 10th gene of a putative operon of 14 genes that was called the imp (impaired in nitrogen fixation) locus. Several highly similar gene clusters of unknown function are present in Pseudomonas aeruginosa, Vibrio cholerae, Edwardsiella ictaluri, and several other animal pathogens. Homology studies indicate that several genes of the imp locus are involved in protein phosphorylation, either as a kinase or dephosphorylase, or contain a phosphoprotein-binding module called a forkhead associated domain. Other proteins show similarity to proteins involved in type III protein secretion. Two dimensional gel electrophoretic analysis of the secreted proteins in the supernatant fluid of cultures of RBL5523 and RBL5787 showed the absence in the mutant strain of at least four proteins with molecular masses of approximately 27 kDa and pIs between 5.5 and 6.5. The production of these proteins in the wild-type strain is temperature dependent. Sequencing of two of these proteins revealed that their first 20 amino acids are identical. This sequence showed homology to that of secreted ribose binding proteins (RbsB) from Bacilus subtilis and V. cholerae. Based on this protein sequence, the corresponding gene encoding a close homologue of RbsB was cloned that contains a N-terminal signal sequence that is recognized by type I secretion systems. Inoculation of RBL5787 on pea plants in the presence of supernatant of RBL5523 caused a reduced ability of RBL5787 to nodulate pea and fix nitrogen. Boiling of this supernatant before inoculation restored the formation of effective nodules to the original values, indicating that secreted proteins are indeed responsible for the impaired phenotype. These data suggest that the imp locus is involved in the secretion to the environment of proteins, including periplasmic RbsB protein, that cause blocking of infection specifically in pea plants. PMID- 12580284 TI - Intergeneric transfer of chromosomal and conjugative plasmid genes between Ralstonia solanacearum and Acinetobacter sp. BD413. AB - Conjugative transfer of a broad-host range plasmid and transformation-mediated transfer of chromosomal genes were found to occur at significant frequencies between Ralstonia solanacearum and Acinetobacter sp. in planta. These intergeneric gene transfers are related to the conditions provided by the infected plant, including the extensive multiplication of these two bacteria in planta and the development of a competence state in Acinetobacter sp. Although interkingdom DNA transfer from nuclear transgenic plants to these bacteria was not detectable, plants infected by pathogens (e.g., Ralstonia solanacearum) and co-colonized by soil saprophyte bacteria (e.g., Acinetobacter sp.) can be considered as potential "hot spots" for gene transfer, even between phylogenetically remote organisms. PMID- 12580283 TI - BacS: an abundant bacteroid protein in Rhizobium etli whose expression ex planta requires nifA. AB - Rhizobium etli CFN42 bacteroids from bean nodules possessed an abundant 16-kDa protein (BacS) that was found in the membrane pellet after cell disruption. This protein was not detected in bacteria cultured in tryptone-yeast extract. In minimal media, it was produced at low oxygen concentration but not in a mutant whose nifA was disrupted. N-terminal sequencing of the protein led to isolation of a bacS DNA fragment. DNA hybridization and nucleotide sequencing revealed three copies of the bacS gene, all residing on the main symbiotic plasmid of strain CFN42. A stretch of 304 nucleotides, exactly conserved upstream of all three bacS open reading frames, had very close matches with the NifA and sigma 54 consensus binding sequences. The only bacS homology in the genetic sequence databases was to three hypothetical proteins of unknown function, all from rhizobial species. Mutation and genetic complementation indicated that each of the bacS genes gives rise to a BacS polypeptide. Mutants disrupted or deleted in all three genes did not produce the BacS polypeptide but were Nod+ and Fix+ on Phaseolus vulgaris. PMID- 12580285 TI - Salicylic acid inhibits indeterminate-type nodulation but not determinate-type nodulation. AB - LCOs (lipochitin oligosaccharides, Nod factors) produced by the rhizobial symbiote of Vicia sativa subsp. nigra (vetch, an indeterminate-type nodulating plant) are mitogenic when carrying an 18:4 acyl chain but not when carrying an 18:1 acyl chain. This suggests that the 18:4 acyl chain specifically contributes to signaling in indeterminate-type nodulation. In a working hypothesis, we speculated that the 18:4 acyl chain is involved in oxylipin signaling comparable to, for example, signaling by derivatives of the 18:3 fatty acid linolenic acid (the octadecanoid pathway). Because salicylic acid (SA) is known to interfere with oxylipin signaling, we tested whether nodulation of vetch could be affected by addition of 10(-4) M SA. This concentration completely blocked nodulation of vetch by Rhizobium leguminosarum bv. viciae and inhibited the mitogenic effect of 18:4 LCOs but did not affect LCO-induced root-hair deformation. SA did not act systemically, and only biologically active SA derivatives were capable of inhibiting nodule formation. SA also inhibited R. leguminosarum bv. viciae association with vetch roots. In contrast, addition of SA to Lotus japonicus (a determinate-type nodulating plant responding to 18:1 LCOs) did not inhibit nodulation by Mesorhizobium loti. Other indeterminate-type nodulating plants showed the same inhibiting response toward SA, whereas SA did not inhibit the nodulation of other determinate-type nodulating plants. SA may be a useful tool for studying fundamental differences between signal transduction pathways of indeterminate- and determinate-type nodulating plants. PMID- 12580286 TI - E. P. Pope Memorial Award to Dr. Gavin Lee Meerdink. PMID- 12580287 TI - Application of the Neospora caninum IgG avidity ELISA in assessment of chronic reproductive losses after an outbreak of neosporosis in a herd of beef cattle. AB - Point-source infections are most likely the cause for Neospora caninum-induced abortion outbreaks in cattle, whereas an increased annual abortion rate may be a consequence of vertical transmission. The aims of the present study were to examine the reproductive effects of neosporosis in a beef herd for 3 years, after a point-source outbreak and to use IgG avidity serology to examine the chronicity of infections and patterns of transmission. During the study, 76-78% of animals were seropositive for N. caninum. The pregnancy rate varied from 88% to 94%, without any reduction in the pregnancy rate of seropositive cows compared with seronegative cows. The annual abortion rate was 2.5-5.5%, and all but 1 abortion occurred in seropositive dams. The efficiency of vertical transmission was estimated to be 85%. Several calves, born to seronegative dams, were seropositive at 6-13 months of age, indicating a 22% mean annual rate of horizontal transmission. The mean avidity in seropositive cows increased from 30 during the initial outbreak to 74 after 3 years. The mode of IgG avidity was 21-40 during the initial abortion outbreak, 41-60 after 1 year, and 61-80 after 2 and 3 years. The results reveal high annual rates of both vertical and horizontal transmission of N. caninum in a herd of beef cows and provide further validation on the ability of the N. caninum IgG avidity ELISA to accurately assess the chronicity of infection. PMID- 12580288 TI - Comparison of a serum indirect fluorescent antibody test with two Western blot tests for the diagnosis of equine protozoal myeloencephalitis. AB - A serum indirect fluorescent antibody test (IFAT) was compared with a Western blot (WB) and a modified Western blot (mWB) for diagnosis of equine protozoal myeloencephalitis (EPM). Using receiver-operating characteristic (ROC) analysis, the area under the curve of the IFAT was greater than the areaunder the curves of the WB and the mWB (P = 0.025 and P = 0.044, respectively). There was no statistically significant difference between the areas under the curves of the WBs (P > 0.05). On the basis of an arbitrarily chosen cut-off titer for a positive test result of 1:80 for the IFAT and interpreting weak positive WB results as positive test results, the sensitivities and 95% confidence intervals (CI) of all 3 tests were identical and equal to 88.9% (51.8-99.7%). The specificities and 95% CIs of the IFAT, WB, and mWB test were 100% (91-100%), 87.2% (72.6-95.7%), and 69.2% (52.4-83%), respectively. The overall accuracy of the IFAT was shown to be better than that of the WBs and, therefore, the test has potential for use in the diagnosis of EPM caused by Sarcocystis neurona. PMID- 12580289 TI - Comparison of the 5.8S rRNA gene and internal transcribed spacer regions of trichomonadid protozoa recovered from the bovine preputial cavity. AB - Sequence analysis of the 5.8S rRNA gene and the internal transcribed spacer regions (ITSRs) was used to compare trichomonadid protozoa (n = 39) of varying morphologies isolated from the bovine preputial cavity. A multiple sequence alignment was performed with bovine isolate sequences and other trichomonadid protozoa sequences available in GenBank. As a group, Tritrichomonasfoetus isolates (n = 7) had nearly complete homology. A similarity matrix showed low homology between the T. foetus isolates and other trichomonads recovered from cattle (<70%). Two clusters of trichomonads other than T. foetus were identified. Eighteen isolates comprised 1 group. These isolates shared >99% homology among themselves and with Pentatrichomonas hominis. The other non-T. foetus cluster (n = 14) did not exhibit a high degree of homology (<87%) with other bovine isolates or any of the trichomonad sequences available in GenBank. The sequence homology among isolates in that cluster was >99%, except for 1 isolate that varied from the others in both ITSRs (approximately 2% dissimilarity). Sequence analysis of the 5.8S rRNA gene and ITSRs was useful for comparing trichomonadid protozoa isolated from the bovine preputial cavity and demonstrated that 2 distinct types of trichomonads constituted the non-T. foetus isolates recovered from the bovine preputial cavity. PMID- 12580290 TI - An estimated prevalence of Johne's disease in a subpopulation of Alabama beef cattle. AB - The objective of this study was to estimate the overall prevalence of animals that were infected with Mycobacterium avium ssp. paratuberculosis in a subpopulation of Alabama beef cattle. This was determined using a commercial enzyme-linked immunosorbent assay (ELISA) for the detection of M. avium ssp. paratuberculosis-specific antibodies in serum. Serum was collected from 79 herds that were participating in the Alabama Brucellosis Certification program. A total of 2,073 beef cattle were randomly tested by selecting 30 animals per herd in herds greater than 30 and selecting all animals in herds 30 and less for testing. It has been estimated that the commercial ELISA test used has a 60% sensitivity and a 97% specificity. Of the 79 herds tested, 29 herds were seronegative, 24 herds had 1-2 positive animals, and 26 herds had 3 or more seropositive animals. The average number of infected animals per positive herd was 3.3. In addition, a calculated minimum of 53.5% of the herds were identified as Johne's positive herds with a 95% confidence level. Of the total number of animals tested, 8.0% (166/2,073) of them were positive by the ELISA. After adjustments for test sensitivity and specificity and the proportion of animals sampled per herd, the true prevalence was calculated to be 8.75%. These data suggest that approximately 50% of the herds are infected with M. avium ssp. Paratuberculosis, and the overall prevalence of infection in Alabama beef cattle is approximately 8%, which correlates with other previously published regional estimates. PMID- 12580291 TI - Antimicrobial susceptibility of Riemerella anatipestifer isolated from ducks and the efficacy of ceftiofur treatment. AB - The in vitro susceptibilities of 50 field isolates of Riemerella anatipestifer from ducks to ceftiofur and 16 other commonly used antimicrobials were determined. The MIC90 values (MIC refers to minimum inhibitory concentrations) for the antimicrobials used in this study are as follows: penicillin was 16 microg/ml; ceftiofur was 32 microg/ml; cephalothin, chloramphenicol, flumequine, and kanamycin were 64 microg/ml; nalidixic acid, nitrofurantoin, and sulfamethoxazole were 128 microg/ml; amikacin, ampicillin, gentamicin, lincomycin, spectinomycin, streptomycin, tetracycline, and trimethoprim were > or = 256 microg/ml. The therapeutic efficacy of ceftiofur against a highly lethal experimental R. anatipestifer infection in ducks was also evaluated. All experimental ducks were infected through the infraorbital sinus with 1 ml of 9 x 10(9) CFU of R. anatipestifer. Ceftiofur (0, 0.25, 0.5, 1, and 2 mg/kg) was injected subcutaneously 5 hours after infection. A single dose of 2 mg/kg resulted in 73% survival as compared with 10% survival in the infected, but untreated controls. PMID- 12580292 TI - Isolation of H3N2 swine influenza virus in South Korea. AB - Swine influenza is a significant respiratory disease causing occasional reproductive problems in naive swine herds. Although different subtypes of swine influenza virus (SIV) have been implicated in clinical outbreaks of swine influenza in Asian countries, no virus isolation has been made to identify SIV of subtypes other than the H1N1 subtype in the Korean swine population. In December 1998, an outbreak of acute respiratory disease was identified in a commercial swine farm located in the Kyunggi province of South Korea. A causative agent, which agglutinated rooster red blood cells, was detected from the lungs of 3 piglets from the index herd and was determined to be type A influenza virus using a commercial influenza virus typing kit. Hemagglutination activity (HA) of the isolates was completely inhibited by a swine antiserum against a recent US H3N2 SIV isolate (A/Sw/IA/41305/1998) but not by H1N1 swine antiserum (A/Sw/IA/1979). Reverse transcription-polymerase chain reaction (RT-PCR) revealed all 3 isolates were H3 SIV subtypes. Sequence analysis of hemagglutinin gene PCR products supported the belief that the Korean H3 SIV isolates were genetically similar to the known mammalian H3 influenza viruses. This is the first report on a clinical outbreak of swine influenza caused by the H3N2 virus in Korea. PMID- 12580293 TI - Detection of classical swine fever virus in boar semen by reverse transcription polymerase chain reaction. AB - A seminested reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of classical swine fever virus (CSFV) in semen. Five boars were inoculated intranasally with CSFV isolate propagated in PK15 cells. Two boars inoculated with the supernatant of noninfected PK15 cells were kept as controls. Semen and serum samples were collected twice weekly for 63 days postinoculation (dpi). Samples were tested for the presence of antibodies to CSFV by an enzyme-linked immunosorbent assay and for the presence of CSFV nucleic acid by seminested RT-PCR. Antibodies to CSFV could be detected as early as 7 dpi in 1 boar, and all 5 infected boars were found positive by 14 dpi. CSFV from boar semen was infrequently identified by virus isolation compared with seminested RT PCR. CSFV nucleic acid was detected in semen by seminested RT-PCR as early as 7 dpi in 3 infected boars and persistently thereafter in all 5 infected boars until 63 dpi. When separated fractions of CSFV-contaminated semen were analyzed by the seminested RT-PCR, the CSFV nucleic acid was detected mainly in seminal fluid and occasionally in nonsperm cells. CSFV antigen was also detected in nonsperm cells from semen smear by immunohistochemistry. Thus, infection via semen, specially through CSFV-infected seminal fluid, seems to be a major route of transmission of CSFV. PMID- 12580294 TI - Thin layer chromatography convulsant screen extended by gas chromatography-mass spectrometry. AB - Acute onset convulsive disorders in the canine may result from exposure to a variety of toxicants including strychnine, insecticides, metaldehyde, zinc phosphide, methylxanthines, drugs of abuse, bromethalin, and the tremorgenic mycotoxins (roquefortine and penitrem A). Although several of the above can be identified in a single gas chromatography-mass spectrometry (GC-MS) screen most have to be determined by separate tests. This report describes a modification of the strychnine extraction procedure, which allows thin layer chromatographic (TLC) identification of strychnine, bromethalin, roquefortine, and penitrem A in suspect baits, stomach contents or vomitus, and extends the identification to a wide variety of drugs, pesticides, and environmental contaminants by GC-MS. Samples were mixed with base, extracted into CH2Cl2 and the organic fraction back extracted with acid. The organic fraction (neutrals) was purified by gel permeation chromatography (GPC) and analyzed by TLC to determine penitrem A and bromethalin. The acidic aqueous fraction was adjusted to pH > 9 and extracted into CH2Cl2. The resulting CH2Cl2 layer (bases) was then analyzed by TLC to determine strychnine and roquefortine. The organic basic and neutral fractions were recombined with a late eluting GPC fraction and analyzed by GC-MS. Of 312 samples analyzed by TLC from 1995 to 2001, 35 were positive for strychnine alone, 58 were positive for both roquefortine and penitrem A, 4 were positive for roquefortine alone, and 1 was positive for bromethalin. None of the samples were positive for penitrem A alone. Samples negative by TLC were analyzed by the GC-MS extended procedure since mid-1999, and 14 have shown positive for a wide variety of compounds with convulsant activity. PMID- 12580295 TI - Caprine herpesvirus-2-associated malignant catarrhal fever in white-tailed deer (Odocoileus virginianus). AB - A subacute disease presenting primarily as alopecia and weight loss occurred in 2 white-tailed deer (Odocoileus virginianus) on farms in Minnesota and in Texas. A presumptive diagnosis of malignant catarrhal fever (MCF) was made on the basis of histological lesions. Antibody against an epitope conserved among the MCF group viruses was detected in the serum of both deer. DNA samples from the deer were subjected to a variety of PCR amplifications. Alignment of the amplified sequences from the diseased animals revealed that they were 100% identical to each other and to the same DNA fragment from the newly recognized member of the MCF virus group endemic in domestic goats (Capra hircus), provisionally named caprine herpesvirus 2 (CpHV-2). A seroprevalence survey from one of the deer farms showed a high rate of subclincal infection in the deer population. This study provides further confirmation that CpHV-2 is a pathogen, at least for deer, and emphasizes the risk of loss from MCF when mixing cervids with goats. PMID- 12580296 TI - Identification and characterization of fowlpox virus strains using monoclonal antibodies. AB - The use of 2 monoclonal antibodies (MAbs), P1D9 and P2D4, which recognize different fowlpox virus (FPV) antigens, for the identification and characterization of FPV strains was evaluated. Initially, the MAbs were used in conjunction with a dot blot assay that enabled FPV to be differentiated from the avian herpesvirus, infectious laryngotracheitis virus. Confirmation of the specificity of these MAbs was provided by the demonstration that only FPV antigens were recognized by a combination of both antibodies when used for immunoblotting proteins contained in various avipoxviruses. Later, an antigenic characterization of 11 FPV field isolates, 6 FPV vaccine strains, and 3 pigeonpox virus vaccines was performed by Western blotting with the individual MAbs. Whereas MAb P2D4 consistently recognized a protein with an apparent molecular weight of 60 kD, there was variability in the size of the antigen that was immunoreactive with the other MAb. For example, MAb P1D9 recognized an antigen of apparent molecular weight of 46 kD in all vaccine strains except 2 of FPV origin. In these exceptions, either only a 39-kD or both a 42- and 46-kD protein were immunoreactive. As for the field isolates, a 39-kD antigen was recognized in 8 of them, whereas a 42-kD antigen was detected in the remaining 3. Therefore, the more extensive immunoblotting technique may facilitate FPV strain differentiation, whereas routine diagnosis of fowlpox could be accomplished by using the MAb-based dot blot assay. PMID- 12580297 TI - Dermal malignant melanoma in a horse with multifocal pancytokeratin expression. AB - The current report describes a malignant melanoma in the dermis of a 13-year-old bay Thoroughbred mare. Microscopic examination revealed that tumor cells were arranged in cords and packets within an abundant collagenous stroma containing scattered myxomatous foci. Tumor cells stained positively for S-100, neuron specific enolase, and vimentin and some contained melanin granules. Some clusters of tumor cells were also positive for pancytokeratin. Expression of epithelial cell markers has been described in small numbers of human melanomas but has not been reported previously in equine melanomas. PMID- 12580298 TI - Encephalitozoon cuniculi placentitis and abortion in a quarterhorse mare. AB - Encephalitozoon cuniculi is a microsporidial parasite, which has rarely been reported to cause placentitis in animals. A late-term aborted fetus and placenta from a Quarterhorse were presented to the Livestock Disease Diagnostic Center, University of Kentucky, for diagnostic examination. There was a necrotizing placentitis, with distension of many chorionic epithelial cells by intracytoplasmic vacuoles containing 1-2-microm-diameter, elongated, gram positive organisms. The organisms were identified as E. cuniculi by electron microscopy and by polymerase chain reaction using primers to microsporidial ribosomal DNA. Joints of the fetus were swollen, with gross and microscopic lesions of synovitis; however, E. cuniculi DNA was not detected. PMID- 12580299 TI - Malignant granulosa-theca cell tumor in a two-year-old Miniature Horse. AB - A 2-year-old female Miniature Horse that presented with a history of progressive weight loss, depression, and diarrhea was diagnosed at necropsy with a highly malignant abdominal neoplasm involving the left ovary, kidneys, adrenal glands, intestines, and various abdominal and thoracic lymph nodes. Microscopic examination of these masses revealed large pleomorphic cells that stained positive for vimentin and inhibin and negative for epithelial membrane antigen and placental alkaline phosphatase. Ultrastructural examination of the cells revealed a high nucleocytoplasmic ratio and indented euchromatic nuclei with large nucleoli. Based on the gross, microscopic, immunohistochemical, and ultrastructural features, the neoplasm was identified as a malignant granulosa theca cell tumor, a rare neoplasm in young horses. PMID- 12580300 TI - Chronic hyperplastic and neoplastic cutaneous lesions (Marjolin's ulcer) in hot brand sites in adult beef cattle. AB - Linear exophytic cutaneous lesions (brand keratomas) are a chronic sequel to hot iron branding in a small proportion of beef cattle in the western United States. Rarely, brand keratomas progress to form large ulcerated masses. Samples of chronically thickened skin were collected from 8 adult cattle with hot-brand lesions and from 2 cattle with ulcerated masses at brand sites. Cutaneous thickening was attributable to abrupt transition from normal haired skin to regular epidermal hyperplasia with marked orthokeratotic hyperkeratosis, acanthosis, hypopigmentation, and loss of adnexae. Epithelial atypia was absent. Normal dermal collagen was replaced by mature granulation tissue containing islands of dense hyalinized collagen. Two cows, aged 5 and 13 years, developed large, slow-growing squamous cell carcinomas at brand sites. Malignancy in branded skin is a rare complication of hot-iron branding in cattle and may arise because of malignant transformation of brand keratomas. PMID- 12580301 TI - Papular dermatitis induced in guinea pigs by the biting midge Culicoides sonorensis (Diptera: Ceratopogonidae). AB - Histological, ultrastructural, and virological examinations were performed on abdominal skin from guinea pigs after a blood meal by colony-bred biting midges, Culicoides sonorensis. Small, superficial, cutaneous, crateriform ulcers with necrosis of superficial dermis developed at feeding sites and healed within 24-48 hours. Animals developed nonpruritic erythematous papules 5 days after feeding that persisted until the study ended at 12 days after feeding. Papules corresponded histologically to foci of epidermal hyperplasia and superficial interstitial dermatitis with intraepidermal micropustules and scattered intraepidermal polykaryons. The principal ultrastructural changes were spongiosis in germinal epithelium and neutrophilic-histiocytic exocytosis. No viral agents or broken mouthparts were identified in lesions. The dermatitis may represent a host reaction to persisting insect salivary secretion and should be considered as an additional consequence of blood feeding in future studies involving biting midges. PMID- 12580302 TI - Detection of bovine leukemia virus in blood and milk by nested and real-time polymerase chain reactions. AB - Concerns about retroviruses in livestock and products derived from them have necessitated the development of tests to detect the bovine leukemia virus (BLV) in blood and milk from cattle. Dairy cattle (n = 101) from 5 different geographical areas were used for this study. A nested polymerase chain reaction (PCR) identified 98% of BLV seropositive cattle (n = 80) from blood and 65% from milk, whereas real-time PCR detected 94% of BLV seropositive cattle from blood and 59% from milk. Bovine leukemia virus was also detected by PCR in approximately 10% of seronegative cattle (n = 21), most likely because of early detection before seroconversion. PMID- 12580303 TI - Malignant ovarian teratoma in a red-eared slider (Trachemys scripta elegans). AB - A 7-year-old intact female red-eared slider (Trachemys scripta elegans) presented with anorexia of 2-3-weeks duration. Coelomic radiographs demonstrated 3 shelled eggs in the right cavity. Hormone therapy failed to induce oviposition. Exploratory laparotomy revealed 3 calcified eggs in the right oviduct and a large mass obliterating the left ovary. The large, multilobulate mass had scattered, firm, white nodules and weighed 450 g. Histopathology revealed a malignant teratoma composed of immature embryonaland extraembryonal elements derived from all 3 germ layers with multifocal epithelial anaplasia. This is the first report of a teratoma in chelonian gonadal tissue. Teratomas and reptilian gonadal neoplasia are reviewed. PMID- 12580305 TI - Biology and pathology of the Weaver mutant mouse. PMID- 12580304 TI - Introduction. Dopamine and Parkinson's disease. PMID- 12580306 TI - Histochemical properties of intrastriatal mesencephalic grafts. PMID- 12580307 TI - Structural correlates of process outgrowth and circuit reconstruction. PMID- 12580308 TI - Neurochemical indices of functional restoration. PMID- 12580309 TI - Behavioral recovery of functional responses. PMID- 12580310 TI - Directions for future research. PMID- 12580317 TI - Tumor necrosis factor-alpha -308 gene polymorphism is associated with synthetic hemodialysis graft failure. AB - BACKGROUND: Progressive venous stenosis mediated, in part, by inflammatory cytokines is a major cause of synthetic hemodialysis graft failure. A tumor necrosis factor-alpha (TNF-alpha) gene polymorphism (G to A, position -308) has been shown to increase plasma cytokine levels and severity of diseases with an underlying inflammatory component. METHODS: We genotyped 67 patients with synthetic polytetrafluoroethylene (PTFE) grafts and examined the association of the high-(AA or GA) and low- (GG) production TNF-alpha-08 genotypes with the rate of graft failures/thrombosis and graft survival. RESULTS: Hemodialysis patients with the high-production TNF-alpha genotypes had a significantly increased rate of PTFE graft failure at 90 days (37.2% versus 14%) and 1 year (62.8% versus 34.4%) after graft placement compared with patients with the low-production genotype (respectively). Hemodialysis patients with the high-production TNF-alpha genotypes had significantly lower cumulative PTFE graft survival at 1 year (29.4% +/- 11.1% versus 71.2 +/- 6.8%) and 2 years (22.1% +/- 10.5% versus 48.2 +/- 8.1%) compared with patients with the low-production genotype (respectively). Patients with the A allele had approximately twice the mean thrombosis rate compared with those who had the low-production TNF-alpha genotype (3.3 +/- 0.8 versus 1.7 +/- 0.4 thromboses/patient/year, respectively; mean +/- SEM, p < .05). CONCLUSIONS: These data suggest that the TNF-alpha -308 A allele is associated with increased PTFE graft thrombosis and failure in hemodialysis patients. PMID- 12580318 TI - Effects of off-pump coronary artery bypass grafting on patient outcome. AB - BACKGROUND: Cardiopulmonary bypass (CPB) is associated with postoperative myocardial stunning, hypothermia, formation of microemboli, and systemic inflammatory response syndrome, all of which may prolong recovery from coronary artery bypass grafting (CABG) surgery. This study sought to compare outcomes in patients undergoing CABG off pump versus on pump. METHODS: Outcomes, including mortality and several morbidities, were reviewed in 1,623 on-pump patients and 683 off-pump patients. Morbidities assessed included postoperative bleeding, incidence of multiorgan dysfunction, and neurologic complications. Chi-square and t-test analysis were used to determine statistical significance. RESULTS: Mortality was 42% lower in the off-pump group than the on-pump group. Both critical care and total hospital length of stay were significantly shorter in the off-pump group. The incidence of postoperative bleeding requiring transfusion or a return to the operating room was reduced by 29% in the off-pump group and the incidence of multiorgan dysfunction was reduced by 31%. The off-pump patients also presented a significantly lower incidence of cerebral vascular accidents and seizures than on-pump patients. CONCLUSIONS: We conclude that there is an association between improved patient outcome and off-pump CABG surgery. The outcomes of this study show a statistically significant decrease in mortality, critical care length of stay, total hospital stay, incidence of bleeding requiring transfusion or return to the operating room, amount of blood transfused, incidence of multiorgan dysfunction, cerebral vascular accidents, and seizures in off-pump patients when compared with on-pump patients. Such results support the use of myocardial revascularization off pump as an alternative to CABG surgery on pump. CABG surgery off pump may allow a better postoperative clinical course in patients who are candidates for the procedure. PMID- 12580319 TI - Overexpression of the complementary DNA for human glutamine:fructose-6-phosphate amidotransferase in mesangial cells enhances glucose-induced fibronectin synthesis and transcription factor cyclic adenosine monophosphate-responsive element binding phosphorylation. AB - Hyperglycemia-induced alterations in mesangial cell function and extracellular matrix protein (ECM) accumulation are seen in diabetic glomerulopathy. The hexosamine biosynthesis pathway (HBP) is implicated in mediating several metabolic effects of high glucose (HG) in cells. This pathway converts fructose-6 phosphate to glucosamine (GlcN)-6-phosphate by the rate-limiting enzyme glutamine:fructose-6-phosphate amidotransferase (GFA). We have previously shown that metabolism of glucose through the HBP regulates the effects of glucose on ECM (fibronectin) synthesis and transcription factor (cyclic adenosine monophosphate-responsive element binding [CREB]) phosphorylation in SV-40 transformed rat kidney mesangial cells. UDP-N-acetyl-GlcN is the end product of the HBP and serves as a precursor for O-linked serine/threonine glycosylation of cytoplasmic and nuclear proteins. Here we show that culturing mesangial cells in HG and GlcN increases the level of O-N-acetylglucosamine in several cytoplasmic and nuclear proteins. Inhibition of O-glycosylation by benzyl-2-acetamido-2-deoxy alpha-D-galactopyranoside blocks both HG and GlcN-induced fibronectin synthesis and CREB phosphorylation. To further support the hypothesis that the HBP mediates HG-induced ECM synthesis, a complementary deoxyribonucleic acid (DNA) for human GFA was stably expressed in mesangial cells. Mesangial and GFA-overexpressing cells were cultured in 5 to 25 mM glucose for 48 hours. GFA-overexpressing cells were more sensitive to glucose as they demonstrated increases in fibronectin and CREB phosphorylation at lower glucose concentrations than seen In control cells. In addition, the response to 25 mM glucose for both proteins was increased in GFA when compared with controls. There is no difference in DNA synthesis and cellular adenosine triphosphate levels between the two cell lines. These results suggest that the HBP is a glucose sensor and mediator of the effects of hyperglycemia in the diabetic mesangium. PMID- 12580320 TI - Utility of hepatic phosphorus-31 magnetic resonance spectroscopy in a rat model of acute liver failure. AB - The ability to document the extent of hepatic injury and predict the outcome of fulminant hepatic failure would be helpful in identifying those patients who might benefit from liver transplantation. The aim of the present study was to determine whether in vivo phosphorus-31 magnetic resonance spectroscopy (31P MRS) accurately assesses the severity of liver damage and is of prognostic value in a D-galactosamine (D-galN)-induced model of acute liver failure. Adult male Sprague Dawley rats (n = 36) received an intraperitoneal dose of D-galN (1.0 g/kg), and MRS examinations were performed at peak (48 hours) and in subsequent experiments, just prior to peak (30 hours) hepatic injury. Rats not exposed to D-galN served as controls. The concentration of hepatic phosphorylated metabolites decreased in proportion to the severity of liver injury at 48 hours. Significant correlations were detected between hepatic adenosine triphosphate (ATP) and serum aspartate aminotransferase, bilirubin, and percentage of hepatocyte necrosis identified histologically (r = -.91, -.74, and -.92, respectively; p < .001). Prior to peak hepatic injury (30 hours), 31P MRS was able to predict with 100% accuracy those rats that would survive (ATP > 2.3 mM) and those that would not (ATP < 1.5 mM). When an intermediate cutoff value of 2.0 mM was selected, ATP levels were able to correctly predict survival and death with 80% and 60% accuracy, respectively. These findings indicate that hepatic ATP levels as measured by 31P MRS provide a noninvasive indication of the severity of liver damage and serve as a useful prognostic indicator of outcome in this model of acute liver failure. PMID- 12580321 TI - Effect of desirable fasting triglycerides on the postprandial response to dietary fat. AB - BACKGROUND: The National Cholesterol Education Program (NCEP) recently revised the "desirable" fasting triglyceride (TG) to < 150 mg/dL, and levels exceeding 200 mg/dL are defined as "high." METHODS: To evaluate the postprandial response to dietary fat, 50 studies were conducted in nonobese, normocholesterolemic subjects. Following an overnight fast, subjects consumed an oral fat load (70 g/m2), and postprandial triglyceride (ppTG) measurements were assessed at 2, 4, 6, and 8 hours. Subjects were divided by fasting TG cutpoints of 100 and 150 mg/dL. RESULTS: The prevalence of ppTG samples exceeding 200 mg/dL was significantly lower with fasting TG < 100 mg/dL (n = 116) compared with TG 100 to 150 mg/dL (n = 56) (8% versus 25%; p = .004, chi-square analysis). In addition, fasting TG < 100 mg/dL (n = 29) was associated with a reduced mean 4-hour peak ppTG level compared with fasting TG > 100 mg/dL (n = 21) (125 mg/dL versus 249.8 mg/dL; p < .0001). Multiple linear regression analysis identified fasting TG as the most important determinant of the postprandial response after adjustment for other covariates (p = .0005). CONCLUSIONS: Because ppTG-rich lipoproteins contribute to coronary heart disease risk, fasting TG < 100 mg/dL may be a more desirable cutpoint than fasting TG < 150 mg/dL in coronary heart disease risk factor assessment. PMID- 12580322 TI - Psychoanalysis in the culture. PMID- 12580323 TI - The theory of psychoanalysis: vicissitudes of its evolution. PMID- 12580324 TI - Transference as defense. PMID- 12580325 TI - Freud's encounter with cocaine. PMID- 12580326 TI - Magic and the aesthetic illusion. AB - The aesthetic illusion is the subjective experience that the content of a work of art is reality. It has an intrinsic relation to magic, an intrapsychic maneuver oriented toward modification and control of the extraspyschic world, principally through ego functioning. Magic is ontogenetically and culturally archaic, expresses the omnipotence inherent in primary narcissism, and operates according to the logic of the primary process. Magic is a constituent of all ego functioning, usually latent in later development. It may persist as an archaic feature or may be evoked regressively in global or circumscribed ways. It causes a general disinhibition of instincts and impulses attended by a sense of confidence, exhiliration, and exuberance. The aesthetic illusion is a combination of illusions: (1) that the daydream embodied by the work of art is the beholder's own, the artist being ignored, and (2) that the artistically described protagonist is a real person with a real "world." The first illusion arises through the beholder's emotional-instinctual gratification from his or her own fantasy-memory constellations; the second comes about because the beholder, by taking the protagonist as proxy, mobilizes the subjective experience of the imaginary protagonist's "reality." The first illusion is necessary for the second to take place; the second establishes the aesthetic illusion proper. Both illusions are instances of magic. Accordingly, the aesthetic illusion is accompanied by a heady experience of excitement and euphoria. The relation among the aesthetic illusion, magic, and enthusiasm is illustrated by an analytic case, J. D. Salinger's "The Laughing Man," Woody Allen's Play It Again, Sam, Don Quixote, and the medieval Cult of the Saints. PMID- 12580327 TI - Slow magic: psychoanalysis and "the disenchantment of the world". AB - Freud's work can be situated in terms of the debate between Enlightenment and anti-Enlightenment thought. The attempts of both sides to claim Freud for their position have merit, but they miss the crucial point: namely, that the tension between its Enlightenment and anti-Enlightenment tendencies is what gives Freud's thinking much of its vitality and depth. The task that faces the interpreter is therefore to elucidate that tension and assess the alignment of forces between the two strands in his thought. An examination of the concept of magic in Freud's theory provides an opportunity to pursue this interpretive task. Although the Enlightenment position he often seems to embrace advocates the complete elimination of magic, many "magical" elements remain in his theory and clinical practice. Nor should this situation be deplored, for the ambition to completely exorcise "enchantment" from human experience is one of the misguided excesses of the Enlightenment. The question of an appropriate fate for magic in psychoanalysis is discussed in relation to the vicissitudes of the transference. Finally, the science versus hermenueutics debate is examined in the light of these considerations in an attempt to specify the unique nature of the psychoanalytic experience. PMID- 12580328 TI - Guards at the gate: race, resistance, and psychic reality. AB - Racial difference and similarity in the treatment relationship highlight the question of what is real and apparently psychological in the different racial experiences of patients and therapists. The relationship between racial material and resistance is discussed, as well as the constancy and fluidity of race as intrapsychic content and the distinction psychoanalysts draw between social and psychic reality. Race in the mind, it is argued, is determined by both internal and external factors, and both are important in the analysis of racial material in the transference. PMID- 12580329 TI - Andrea del Sarto rehabilitated: a psychoanalytic emendation. AB - Ernest Jones's "The Influence of Andrea del Sarto's Wife on His Art" (1913) is an early example of psychoanalysis applied to the study of a prominent painter. Greatly influenced by Freud's Leonardo da Vinci and a Memory of His Childhood, Jones gave excessive credence to Vasari's highly prejudicial account of the life of del Sarto, on which the study relied heavily. Jones attempted to account psychologically for the circumstance that del Sarto, though highly skilled and "faultless," was not the equal of the three preeminent masters of the Italian High Renaissance: Leonardo, Michelangelo, and Raphael. Jones's uncritical acceptance of the Vasari biography encouraged him to view Sarto's assumed deficiency as the result of excessive attachment to his wife, a pathological uxoriousness. A contemporary psychoanalytic perspective, with its emphasis on the emotive response of the analyst, requires us to pay attention to the evocative nature of the work of the artist, an approach Jones neglected. In an examination of several paintings, the artist's sensitivity to the position of the spectator is explored, as is the interest in involving the viewer spatially and emotionally. An appreciation for the viewer's position is consistent with a capacity for using projected internal objects for creative purposes. The presence of this capacity suggests a revised view of del Sarto's contribution to art and of his relationship with his wife. PMID- 12580330 TI - Tennessee Williams: the uses of declarative memory in The Glass Menagerie. AB - Tennessee Williams called his first great work, The Glass Menagerie, his "memory play." The situation in which Williams found himself when he began writing the play is explored, as are the ways in which he used the declarative memory of his protagonist, Tom Wingfield, to express and deal with his own painful conflicts. Williams's use of stage directions, lighting, and music to evoke memory and render it three-dimensional is described. Through a close study of The Glass Menagerie, the many uses of memory for the purposes of wish fulfillment, conflict resolution, and resilience are examined. PMID- 12580331 TI - James Joyce and Molly Bloom: reflections on their relationship. AB - The relationship between James Joyce and his memorable creation, Molly Bloom, is explored in relation to Joyce's remarkable creativity and various factors that may have contributed to it. A character forged primarily out of Joyce's perceptions of his wife Nora and memories of his mother, Molly also contains aspects of Joyce's warded-off and wished-for self-representation. A focus on both biographical and dynamic contributions to the creation of Molly helps to illuminate aspects of Joyce's psychology. PMID- 12580332 TI - Flaubert and Madame Bovary: an intimate courtship. AB - The process by which Gustave Flaubert created the character of Emma Bovary is examined, as are various of the author's sources for the heroine and their transformation in the course of composing the novel. Certain aspects of the authors psychic makeup, including his bisexuality, are discussed in this light, as are Flaubert's early traumatic losses and their influence on his way of working. Finally, it is suggested that writing had multiple functions for the author and that the creation of Emma Bovary served as a partial solution to unmet needs. PMID- 12580334 TI - Cavell, Friedman, and objectivity. PMID- 12580333 TI - Catching the ball: the role of play in psychoanalytic treatment. PMID- 12580335 TI - Gap junctions and neuronal injury: protectants or executioners? AB - The authors review concepts and recent experimental observations that relate gap junctional communication to the pathophysiology of neuronal injury, specifically ischemic or traumatic damage. The role played by this type of direct intercellular communication during the progression of the injuries can be conceived to be either detrimental or beneficial, depending on the arguments employed. The data indicate that, far from being a simple matter of judgment, the contribution of gap junctions to cell injury is a complicated phenomenon that depends on the specific insult and network in which it operates. PMID- 12580336 TI - Cytokines: powerful regulators of glial cell activation. AB - It is now clear that cytokines function as powerful regulators of glial cell function in the central nervous system (CNS), either inhibiting or promoting their contribution to CNS pathology. Although these interactions are complex, the availability of animals with targeted deletions of these genes and/or their receptors, as well as transgenic mice in which cytokine expression has been targeted to specific cell types, and the availability of purified populations of glia that can be studied in vitro, has provided a wealth of interesting and frequently surprising data relevant to this activity. A particular feature of many of these studies is that it is the nature of the receptor that is expressed, rather than the cytokine itself, that regulates the functional properties of these cytokines. Because cytokine receptors are themselves modulated by cytokines, it becomes evident that the effects of these cytokines may change dramatically depending upon the cytokine milieu present in the immediate environment. An additional exciting aspect of these studies is the previously underappreciated role of these factors in repair to the CNS. In this review, we focus on current information that has helped to define the role of cytokines in regulating glial cell function as it relates to the properties of microglia and astrocytes. PMID- 12580337 TI - Muscarinic and nicotinic cholinergic mechanisms in the mesostriatal dopamine systems. AB - The striatum and its dense dopaminergic innervation originating in the midbrain, primarily from the substantia nigra pars compacta and the ventral tegmental area, compose the mesostriatal dopamine (DA) systems. The nigrostriatal system is involved mainly in motor coordination and in disorders such as Tourette's syndrome, Huntington's disease, and Parkinson's disease. The dopaminergic projections from the ventral tegmental area to the striatum participate more in the processes that shape behaviors leading to reward, and addictive drugs act upon this mesolimbic system. The midbrain DA areas receive cholinergic innervation from the pedunculopontine tegmentum and the laterodorsal pontine tegmentum, whereas the striatum receives dense cholinergic innervation from local interneurons. The various neurons of the mesostriatal systems express multiple types of muscarinic and nicotinic acetylcholine receptors as well as DA receptors. Especially in the striatum, the dense mingling of dopaminergic and cholinergic constituents enables potent interactions. Evidence indicates that cholinergic and dopaminergic systems work together to produce the coordinated functioning of the striatum. Loss of that cooperative activity contributes to the dysfunction underlying Parkinson's disease. PMID- 12580338 TI - Spreading acidification and depression in the cerebellar cortex. AB - Optical imaging of activity-dependent pH changes using neutral red has revealed a novel form of propagated activity in the cerebellar cortex: spreading acidification and depression (SAD). Evoked by surface stimulation, SAD is characterized by a propagation geometry that reflects the parasagittal architecture of the cerebellum, high speed of propagation across several folia, and a transient depression of the molecular layer circuitry. The properties of SAD differentiate it from other forms of propagating activity in the nervous system including spreading depression and Ca++ waves. Involving several factors, SAD is hypothesized to be a regenerative process that requires a functioning parallel fibers-Purkinje cell circuit, glutamatergic neurotransmission, and is initiated by increased neuronal excitability. Three possible neuronal and glia substrates in the cerebellar cortex could account for the propagation geometry of SAD. Recently, the authors demonstrated that blocking voltage-gated Kv1.1 potassium channels plays a major role in the generation of SAD. This observation has lead to the hypothesis that the episodic and transient disruption in cerebellar function that characterizes episodic ataxia type 1, a Kv1.1 channelopathy, is due to SAD occurring in the cerebellar cortex. PMID- 12580339 TI - Two-pore-Domain (KCNK) potassium channels: dynamic roles in neuronal function. AB - Leak K+ currents contribute to the resting membrane potential and are important for modulation of neuronal excitability. Within the past few years, an entire family of genes has been described whose members form leak K+ channels, insofar as they generate potassium-selective currents with little voltage- and time dependence. They are often referred to as "two-pore-domain" channels because of their predicted topology, which includes two pore-forming regions in each subunit. These channels are modulated by a host of different endogenous and clinical compounds such as neurotransmitters and anesthetics, and by physicochemical factors such as temperature, pH, oxygen tension, and osmolarity. They also are subject to long-term regulation by changes in gene expression. In this review, the authors describe multiple roles that modulation of leak K+ channels play in CNS function and discuss evidence that members of the two-pore domain family are molecular substrates for these processes. PMID- 12580340 TI - The neurobiology of Rett syndrome. AB - Rett syndrome is a neuropsychiatric disorder with onset in early childhood. Loss of-function mutations of the X-linked gene encoding methyl-CpG binding protein 2 (MECP2) are responsible for more than 80% of Rett cases. Despite these recent advances in molecular genetics, little is known about the neurobiology of Rett syndrome and the role of MeCP2 protein in the nervous system. The molecular functions of the MeCP2 protein were primarily studied in nonneuronal cell lines and in vitro systems; MeCP2 binds primarily, but not exclusively, to methylated DNA, and it is thought to regulate gene expression, chromatin composition, and chromosomal architecture. In the brain, MeCP2 appears to be expressed ubiquitously by neurons. There is increasing evidence that the protein is important for maintenance of neuronal chromatin during late development and in adulthood. Levels of MeCP2 expression increase during the course of neuronal differentiation and remain at high levels in the adult brain. Furthermore, genetic studies in mice demonstrated that the selective deletion of the Mecp2 gene in neurons results several weeks later in a Rett-like phenotype, including a reduction in brain weight and neuronal dystrophy. Rett syndrome is not accompanied by a neurodegenerative process and thus perhaps may be amenable to therapeutic intervention at the time of symptom expression. PMID- 12580341 TI - Plasticity of cortical projections after stroke. AB - Ischemic stroke produces cell death and disability, and a process of repair and partial recovery. Plasticity within cortical connections after stroke leads to partial recovery of function after the initial injury. Physiologically, cortical connections after stroke become hyperexcitable and more susceptible to the induction of LTP Stroke produces changes in the distribution and laterality of sensory, motor, and language representations within the brain that correlate with functional recovery. Anatomically, ischemic lesions induce axonal sprouting within local, intracortical projections and long distance, interhemispheric projections. This postischemic axonal sprouting establishes substantially new patterns of cortical connections with de-afferented or partially damaged brain areas. Axonal sprouting after ischemic lesions is induced by a transient pattern of synchronous, low-frequency neuronal activity in a network of cortical areas connected to the infarct. This pattern of neuronal activity that induces axonal sprouting in the adult after ischemic lesions resembles that seen in the developing brain during axonal elongation and synaptogenesis. Thus, stroke induces a process of remapping and reconnection within the adult brain through changes in neuronal activity that may involve a reactivation of developmental programs in areas connected to the infarct. PMID- 12580342 TI - Dystonia: lessons from brain mapping. AB - Functional neuroimaging with positron emission tomography, single photon emission computer tomography, magnetic resonance imaging, and magnetoencephalography have provided powerful tools to elucidate anatomo-functional impairment underlying movement disorders such as dystonic movements. They have revealed that presymptomatic cerebral abnormalities may be a common feature in dystonia whatever the clinical status. Techniques using specific markers have recently focused on the type of receptors that may be dysactivated and on the kind of neurotransmitter that may be dysregulated in dystonia. PMID- 12580343 TI - The neurobiology of depression: perspectives from animal and human sleep studies. AB - This article reviews human and animal studies in the neurobiology of depression. The etiology of the illness, associated neurotransmitter dysregulation, sex steroids, the role of stress, and sleep regulation are discussed. It is suggested that the genesis of depression is related to homeostatic maladaptation that is sexually dimorphic. The authors propose that depressed females are hyperresponsive to stress, whereas depressed males are hyporesponsive to stress. This divergence reflects the exaggeration of naturally occurring differences between males and females, which are most obvious under challenge conditions. The authors conclude that future work in this area should fully evaluate sexual dimorphism, neural plasticity, critical periods, and individual differences in vulnerability. PMID- 12580344 TI - Chronic plantar fasciitis. PMID- 12580345 TI - Management of peroneal nerve injuries associated with knee dislocations. AB - Peroneal nerve palsy after knee dislocation is a serious problem, and neurolysis at the time of knee reconstruction does not always result in return of peroneal nerve function. In light of the current state of microneural surgery, and the potential to reconstruct lengthy nerve defects, this review addresses the appropriate evaluation of peroneal nerve palsy and provides recommendations regarding surgical treatment. PMID- 12580346 TI - Physical and monetary costs associated with autogenous bone graft harvesting. AB - Although autogenous iliac bone is frequently used for bone graft, many well documented complications are associated with this procedure-including chronic pain; nerve, arterial, and ureteral injury; herniation of abdominal contents; sacroiliac joint instability; pelvic fractures; hematoma; and infection. An understanding of the morbidities associated with bone graft harvesting and of the strategies for avoiding them is imperative for surgeons using this grafting source. In addition, although synthetic grafting materials are considered relatively expensive compared with autogenous sources, the majority of physicians are unaware of the actual direct and indirect costs associated with autogenous bone graft harvesting. Contemporary allograft and synthetic grafting composites are being developed to optimize and surpass the native qualities of autogenous sources (ie, osteogenesis, osteoinductivity, osteoconductivity). Careful comparison of the cost of these alternative sources with the physical and monetary costs of autogenous bone graft will undoubtedly make allograft, recombinant, synthetic graft composites the logical choice in the very near future. PMID- 12580347 TI - Varus collapse of comminuted distal femur fractures after open reduction and internal fixation with a lateral condylar buttress plate. AB - Twenty-six comminuted distal femur fractures treated with a lateral condylar buttress plate were followed up until fracture union or implant revision (mean follow-up, 26 months). Mean postoperative angle (medial distal femoral angle immediately after surgery) was 96 degrees, and mean final angle (angle at fracture healing or implant revision) was 91 degrees (P = .06). Distal femoral angle decreased 5 degrees or less in 15 patients, 6 degrees to 10 degrees in 8 patients, and more than 10 degrees in 3 patients. Final distal femoral angle was 90 degrees or less in 13 of the 26 fractures: 38% of these 13 fractures required reconstruction for nonunion, malunion, or knee arthrosis. The lateral condylar buttress plate, which is not a fixed-angle device, allowed more than 5 degrees of varus collapse to occur in 42% of the comminuted distal femur fractures. PMID- 12580348 TI - Intraosseous metacarpal involvement of giant cell tumor of the tendon sheath: report of 2 cases. AB - Intraosseous phalangeal growth of giant cell tumor of the tendon sheath (GCTTS) is an uncommon manifestation of this tumor, but it is well known because of its potentially aggressive local growth. This article details the findings of 2 cases of previously unreported intraosseous metacarpal involvement of GCTTS. Differential diagnosis of a metacarpal lesion with decreased radiodensity on radiographs should include GCTTS. PMID- 12580349 TI - Plantar ganglion cyst associated with stress fracture of the third metatarsal. AB - Ganglion cysts of the foot and ankle occur relatively infrequently. Metatarsal stress fractures occur in a variety of athletes who subject their lower extremities to repetitive loading. In this article, we report the case of a professional football player with a plantar forefoot ganglion cyst associated with a stress fracture of the third metatarsal. After the cyst resolved with aspiration, the stress fracture healed with conservative, nonsurgical treatment. PMID- 12580350 TI - Bilateral carpometacarpal dislocations of the thumb. AB - In this article, we present a case of traumatic bilateral trapeziometacarpal joint dislocations treated successfully with closed reduction and immobilization. We also review the literature, describe a treatment protocol for management of trapeziometacarpal dislocations, and recommend conservative treatment for patients who have such injuries and who meet certain criteria. Conservative treatment includes immediate diagnosis and stabilization after acute reduction and then use of immobilization to maintain stabilization. Failure to maintain closed reduction, acute instability, significant swelling, or delayed presentation warrants fixation, possibly with percutaneous pinning, and then immobilization in a thumb spica cast. Loss of reduction after pinning, or failure to achieve anatomic reduction, requires open reduction, possibly with acute ligamentous reconstruction. PMID- 12580351 TI - Intramedullary fixation of diaphyseal femoral fractures in elderly patients: analysis of outcomes and complications. AB - The efficacy of intramedullary fixation for diaphyseal femoral fractures in young patients has been well documented. There is a paucity of data, however, on the efficacy of intramedullary techniques used in the ever growing elderly population. The purpose of this study was to analyze the outcomes and rate of perioperative complications associated with using intramedullary fixation to treat diaphyseal femur fractures in patients older than 65 years. We retrospectively reviewed the medical records and radiographs of 21 patients (17 men, 4 women) with isolated diaphyseal femoral fractures treated with anterograde intramedullary nailing at our institution. All fractures united. Ten surgical complications occurred in 9 (43%) of the 21 patients, and 7 medical complications occurred in 4 patients (19%); overall, 17 medical or surgical complications occurred in 11 patients (52%). Although more than half of the patients had an intraoperative or postoperative complication, intramedullary fixation is an effective method for achieving union in elderly patients with diaphyseal femoral fractures. PMID- 12580352 TI - Use of a vertical transarticular pin for stabilization of severe ankle fractures. AB - Transarticular pin fixation has been used successfully to stabilize severe ankle fractures. This technique is most commonly used as provisional fixation until internal fixation is appropriate. In addition, transarticular pin fixation can be a supplement in cases involving persistent tibiotalar instability after internal fixation and can provide sole definitive fixation of arthritic and osteoporotic ankles. In this article, we describe the surgical technique, report results, and review when transarticular pin fixation may be appropriate. We believe that all orthopedic surgeons should know this technique--especially those treating cases of complex orthopedic trauma. PMID- 12580353 TI - A 7-year-old boy with left sternoclavicular swelling. AB - This case is presented to illustrate the imaging and clinical findings of a condition of interest to orthopaedic surgeons. The initial findings are noted on this page. The clinical and imaging diagnoses are presented on the following pages. PMID- 12580354 TI - "There is nothing wrong with my teeth." What is informed consent for persons with cognitive impairment? PMID- 12580355 TI - Experience and assessment of pain in individuals with cognitive impairments. AB - The authors review the literature on pain experience and pain assessment in people with cognitive impairments, focusing on individuals with dementia and mental retardation. The impact of cognitive impairments on pain sensation is not well understood, although some observations have been published. For example, research suggests that pain experience can be influenced by neuropathological processes in the brain and memory impairments. Reporting of pain decreases as cognitive impairment increases. In addition, poor verbal skills lead to difficulties in communicating pain. Pain assessment depends primarily on one's ability to describe the dimensions of pain. Individuals with limited ability to report pain can use pain assessment methods that rely on simple cognitive tasks. For individuals who have no ability to report pain, an outside observer must describe the discomfort experienced by interpreting the patient's body language. The authors conclude that further research is needed to develop valid and reliable assessment methods for people with cognitive impairments. PMID- 12580356 TI - Inter-rater reliability of an oral assessment guide for elderly patients residing in a rehabilitation ward. AB - The aim of this study was to test the inter-rater reliability of a revised oral assessment guide (ROAG) for patients residing in a geriatric rehabilitation ward. A consecutive sample of 140 patients was recruited for the study. Oral assessments were performed for 133 newly admitted patients by one registered nurse (RN) during a period of six months. A dental hygienist (DH) carried out 103 oral assessments during the same half-year. For 66 patients, the RN and the DH performed independent assessments. There was an agreement between the RN and the DH in the majority of the independent assessments, except for tongue and teeth/dentures. The percentage agreement exceeded 80 percent. Inter-rater agreement measured by Cohen's Kappa coefficient ranged from moderate to very good and percentage agreement had a range of 58 to 91 percent. The agreement was highest in assessment of voice and swallowing (91%). Assessments of teeth and dentures seemed to be most difficult for the RN to evaluate. ROAG was found to be a clinically useful assessment tool. Additional education and training is needed to improve the reliability of the oral assessments and should include continuous support from a dental hygienist as well as a pictorial manual on how to use the ROAG. PMID- 12580357 TI - The effect of polyol-combinant saliva stimulants on S. mutans levels in plaque and saliva of patients with mental retardation. AB - The effect of chewable saliva-stimulants on Streptococcus mutans levels in dental plaque and paraffin-stimulated whole saliva among participants who were mentally disabled was investigated. Over 64-days, 98 participants chewed one of four saliva-stimulating tablets five times/day. The tablets contained one of the following: xylitol (X) or sorbitol (S), or 1:1 mixtures of xylitol and erythritol (XE) or sorbitol and erythritol (SE). Consumption of xylitol and sorbitol in Groups X and S was 5.4 grams/day/ subject, and of each polyol in Groups XE and SE, consumption was 2.7 g/day/subject. Interproximal dental plaque and stimulated whole saliva were sampled at baseline, at Day 36, and Day 64. There was a statistically significant reduction of S. mutans in plaque and saliva counts in Groups X and XE. The percentage of S. mutans in total streptococci increased significantly in dental plaque in Group S but decreased in the other groups. The results suggest that xylitol-containing saliva stimulants may be more effective than sorbitol-containing products in controlling some caries-associated parameters in people who are mentally disabled. Also a relationship may exist between the pentitol-type xylitol and S. mutans, and erythritol may exert a specific biochemical effect on this organism, although further studies are needed. PMID- 12580359 TI - Use of dental implants in patients with Down syndrome: a case report. AB - Down syndrome is caused by trisomy of the 21st chromosome and is associated with well-described physical and systemic problems. Most people with Down syndrome have some degree of mental retardation as well as malformation of head and neck. Oral structures that are commonly affected include the tongue (macroglossia), abnormalities in the number and shape of teeth, and poor quality (osteoporotic like) of alveolar bone and jaw. These oral malformations as well as a tendency toward poor cooperation in the dental office contribute to the belief among dentists that people with Down syndrome are not good candidates for oral rehabilitation with dental implants. This article describes the use of dental implants in the oral rehabilitation of a 16-year-old boy with Down syndrome. Although more experience is needed before dental implants can be considered a suitable option during oral rehabilitation in people with Down syndrome, this case report shows a promising beginning. PMID- 12580358 TI - Oral disorders in institution-dwelling elderly adults: a graphic representation. AB - This study was conducted to assess the oral health status and dental care needs of elderly adults living in long-term care facilities. Dentists examined 601 elderly adults, living in one of six extended-or intermediate-care facilities, between September 1999 and May 2000. Data from 532 of the subjects were analyzed with descriptive statistics and zero-order inverse polynomials. Using a computer program, the authors compiled summaries of oral health data on individuals and institutional levels. This study suggests that there are numerous unmet dental needs among elderly adults who live in institutions. The CODE index used to assess the oral health of these residents offers a systematic portrayal of oral disorders in terms of severity. As the authors demonstrated, this index can be readily analyzed using zero-order inverse polynomials to summarize collected data into a graphic description, which can be helpful in managing and administrating oral health care interventions in long-term care facilities. PMID- 12580360 TI - Hematologic disorders of children in developing countries. AB - This article outlines a few of the hematologic problems that are particular to developing countries, particularly those of the tropics. Because of globalization and the increasing movement of populations, hematologists in wealthier countries must be aware of the general patterns of hematologic change in the important infectious diseases that are common in developing countries. Their manifestations are protean, and any of these diseases, malaria in particular, may present in ways that are atypical from the standard textbook descriptions. In short, the handling of hematologic disorders in developing or tropical countries is no longer confined to the physicians who work in these countries; these diseases are now part of the work of every hematologist. PMID- 12580361 TI - Thalassemia: current approach to an old disease. AB - This article discusses the approach for recognition, diagnosis, and management of the thalassemias, and reviews new prospects of therapy, focusing mostly on the beta-thalassemias--the more severe and clinically important type, beta thalassemia major is typically treated with regular transfusion and chelation therapy. New strategies for specific therapy including monitoring of iron induced organ damage, fetal hemoglobin augmentation as an alternative for transfusions, bone marrow transplantation offer hope for prevention of complications and better care of the beta-thalassemias. PMID- 12580362 TI - Sickle cell disease. AB - Sickle cell disease is a serious and life threatening disease that affects approximately 1 in 600 African-Americans. Since its first description in 1910 by Herrick, our understanding and treatment of this disease has grown tremendously. Most pediatricians will encounter one or more children with sickle cell disease and should have a thorough understanding of the disease, its complications, and treatment. This article summarizes present day knowledge about the complications and potential treatments for patients with sickle cell disease. PMID- 12580363 TI - Transfusion medicine for the pediatrician. AB - In the next decade, many of the methodologies and research reviewed in this article will become clinical practice, making the transfusion of blood products safer and more universally available than they are today. NAT will be standard and will surely be performed on each unit of product, PCR testing for pathogens will evolve, and the pathophysiology and immunology of transfusion-related events such as TRALI and immunomodulation will be elucidated. New methods of preservation and early detection of contamination will extend the life of blood products. Red blood cell antigens may be attenuated, making safe products available to more patients. Clinical vigilance at the bedside and in the blood bank will remain key areas for transfusion safety. As I have told many a resident and patient, blood is not saline; there are and will remain risks inherent in this commonly used medical therapy. PMID- 12580364 TI - Approach to the bleeding child. AB - Because bruising and bleeding are normal events of childhood, the pediatrician must be able to determine whether a child's symptoms are normal or perhaps indicative of a defective hemostasis. A thorough medical history and physical examination should enable the clinician to choose those patients warranting further evaluation. Rather than referral to a hematologist at that point in time, pediatricians should be quite capable of performing the initial laboratory evaluation and making the correct diagnosis in a majority of cases. PMID- 12580365 TI - Pediatric thrombosis. AB - Although thrombosis is less frequent in children than in adults, it represents a significant source of morbidity and mortality. Multiple factors. both genetic and acquired. contribute to the development of thrombosis in chiidren. Thrombosis in a child warrants investigation of potential underlying prothrombotic conditions. The risk of thrombosis in children with heterozygous deficiencies is not clearly defined, but it appears that children who are heterozygous for more than one risk factor or who have a combination of inherited and acquired defects are at higher risk for thrombosis. Treatment of thrombosis primarily involves a rapidly acting anticoaguiant such as heparin or LMWH to prevent extension, and long-term anticoagulation with warfarin may be instituted to prevent recurrence. Thrombolytic therapy with recombinant tissue plasminogen activator also appears to be safe and effective in children. Prospective and multicenter studies are still needed to clarify the contribution of specific prothrombotic disorders to childhood TE so that evidence-based treatment recommendations can be made. PMID- 12580366 TI - Childhood lead poisoning. AB - Although the average blood lead levels of Americans have markedly declined, a significant number of children remain at risk. This article discusses the mechanisms of lead poisoning and the screening of children for lead poisoning, and the treatment of symptomatic and asymptomatic children. PMID- 12580367 TI - Pediatric oncology: current views and outcomes. AB - The extraordinary explosion of molecular and cellular insights may provide potentially exploitable opportunities to meet the challenge of curing and ultimately preventing all cancer in children. This article describes the various approaches in developing molecular therapy targeted at common childhood cancers. PMID- 12580368 TI - The pediatrician and medical care of the child with cancer. AB - The child's pediatrician is an important resource for families regarding a host of issues such as infections, complications and side effects of chemotherapy, school issues, and psychosocial stressors that are common in families of children with life-threatening illness. This article provides guidance for caring for children with malignancies in the primary care setting. PMID- 12580370 TI - Malignant abdominal masses in children: quick guide to evaluation and diagnosis. AB - A palpable mass in the abdomen of a child is a serious finding. In this article the authors present their single-institution experience of how these malignancies present and their distribution by age and diagnosis. The most common abdominal malignancies diagnosed in the pediatric population include neuroblastoma, Wilms' tumor, hepatoblastoma, lymphoma, and germ cell tumors. This article provides the busy general pediatrician with some guidelines of how to proceed after discovering a suspiciousmass. PMID- 12580369 TI - Emerging cancer-targeted therapies. AB - There is reason to believe that the unfolding revolution in molecular biology and translational research will allow selective targeting of tumor cells, and radically change the way general practitioners and pediatric oncologists treat and follow children with cancer. This article highlights some of the most promising approaches being tested in the field. By learning about the underlying biology, the remaining hurdles, the projected timeline, and the possible impact of new therapies on the practice of pediatric oncology, health care professionals and patients should be better prepared for the future of pediatric oncology. PMID- 12580371 TI - Genetic predisposition and screening in pediatric cancer. AB - Pediatricians are often the health care providers who first detect the signs and symptoms of childhood cancer. Although pediatric malignancies are rare diseases, early diagnosis is an important factor leading to high cure rates of many types of cancers including retinoblastomara, Wilms' tumor, hepatoblastoma, rhabdomyosarcoma. thyroid carcinoma, and other solid tumors. A number of familial cancer syndromes present with childhood cancers that can be recognized or diagnosed by pediatricians. The genetic origins of several syndromes have been elucidated. Genetic testing is not yet available for all of these inherited cancers. A frequently updated list of genetic tests is available at www.genetests.org. The ordering and interpreting of genetic tests, however, is often best done by trained genetic counselors. The pediatrician will play a vital on-going role in following the at-risk child. In many of syndromes discussed, the cost effectiveness of the tests as well as that of any potential intervention needs further study. The role of the subtle genetic polymorphisms in pediatric tumorigenesis. many more of which will undoubtedly be described in the coming years, has not yet been translated into defined needs for interventions. Perhaps in the future it will be possible to understand the additive effect of multiple genetic polymorphisms and to determine genetic profiles of high cancer risk. Until suitable interventions are established, however, the study of genetic variability and cancer will await practical significance. Undoubtedly other major important cancer genes are yet to be discovered and characterized. An additional challenge is the counseling and management of children and adults who have a strong family history of cancer yet who do not have a recognizable syndrome. The role of the primary pediatrician is to recognize the major cancer genetic syndromes, to make appropriate referrals for genetic counseling and testing when indicated, and to ensure that adequate screening tests are being done. PMID- 12580372 TI - Molecular genetic diagnosis of pediatric cancer: current and emerging methods. AB - A wide array of diagnostic tests are available to evaluate molecular abnormalities in pediatric cancer. Classic cytogenetics, FISH, flow cytometry, PCR, and Southern blot analysis are in widespread use throughout pediatric hospitals. Examples of the application of these methods in pediatric cancer diagnosis are reviewed. Newer methods such as CGH, SKY, gene expression microarrays and proteomic methods are under active investigation andwill almost certainly lead to significant advances in our ability to diagnose and treat pediatric cancer. PMID- 12580373 TI - New approaches to hematopoietic cell transplantation in oncology. AB - The use of high-dose chemotherapy followed by autologous HCT and the use of allogeneic HCT in children and adolescents with high-risk ALL, AML, and NBL has successfully improved outcomes. For other diseases, however, the role of HCT in treatment remains a subject of further research. The availability of HCT was significantly expanded by developing alternative graft sources that currently include BM, peripheral blood, and UCB from autologous and allogeneic related or unrelated donors. Progress in autologous HCT has been achieved by the identification of more effective and less toxic preparative regimens and by ex vivo purging of stem cell products. In allogeneic HCT, graft-versus-leukemia or graft-versus-tumor effects are being exploited increasingly to lower relapse rates. In addition, immunomodulation to promote tolerance, as well as allogeneic antitumor reactions have been achieved by antibody therapy, cytokine therapy, or cell-based immunotherapy. Future improvements are likely, as evidenced by promising preliminary results in the development of stem cell collection techniques, in vitro stem cell expansion, and purging techniques of stem cell grafts. The development of less intensive or nonmyeloablative preparative regimens may further reduce regimen-related morbidity and mortality Specific immunotherapy may facilitate tolerance induction in mismatched allogeneic HCT and support allogeneic HCT in the setting of donor-host HLA disparity. Ultimately, advances in cytokine therapy, tumor-specific vaccines, and gene therapy may decrease or even eradicate recurrence of the malignant disease after HCT. PMID- 12580374 TI - Viral proteinases--possible targets of antiviral drugs. AB - Viral infections represent various types of human, veterinary and plant diseases with a significant economic, ethic and demographic impact. Over the years a significant effort has been made to develop various means of prevention and therapy of viral diseases. Proteinases play an important role in the process of virus replication as well as in the pathophysiology of many viral diseases. The aim of this review is to assess the prospects of the application of proteinase inhibitors in antiviral therapy and to characterize viral proteinases of various classes. Six Human immunodeficiency virus (HIV) proteinase inhibitors have been approved for therapeutic use and can serve as examples of prospective application ofproteinase inhibitors to antiviral therapy. PMID- 12580375 TI - Application of cDNA microarray technique to detection of gene expression in host cells infected with viruses. AB - cDNA microarray technique was used to monitor changes in mRNA levels in cells after Hantaan virus (HTNV) infection. The values of the ratio of medians for HTNV and Japanese encephalitis virus (JEV) at the early stage of infection were compared and found similar, suggesting that the same or similar genes are associated with the early events of infection with either virus. The reproducibility of values of the "ratio of medians" for HTNV was examined. We found that applying cluster analysis to the gene expression data groups efficiently together genes with the same function. Therefore, in analyzing the effects of viral infection on host cells by the cDNA microarray technique, clustering data appear to be necessary for gaining biological meaning from a dump of gene expression profiles obtained from virus-infected cells. PMID- 12580376 TI - Polyclonal antibodies to a recombinant coat protein of Potato virus A. AB - Specific mouse antibodies against a recombinant coat protein (CP) of Potato virus A (PVA) were produced. The PVA CP gene was cloned in an expression vector pMPM4omega. After expression in Escherichia coli the presence of the expressed CP was proved by Western blot analysis using polyclonal and monoclonal antibodies (MAbs). The expressed CP was purified by centrifugation in CsCl density gradient or on a sucrose cushion. The production of virus-like particles (VLPs) was proved by electron microscopy. The purified CP was used for preparation of a mouse antiserum which had a titer of 1:1024 in ELISA and reacted specifically in Western blot analysis and indirect plate-trapped antigen ELISA (PTA-ELISA). PMID- 12580377 TI - Complete genomes of two Human hepatitis A virus isolates from China: analysis and comparison with other isolates. AB - Complete sequences of the genomes of two wild type (wt) Human hepatitis A virus (HHAV) isolates, LU38 and LY6 from China were determined and compared with those of wt HHAV isolates AH1, AH2, AH3, FH1, FH2, FH3, GBM, HM175, LA and MBB. The genomes of both LU38 and LY6 consisting of 7477 nucleotides (nts) contained a 5' non-translated region (5'-NTR, 733 nts), an open reading frame (ORF, 6681 nts), and a 3'-NTR (63 nts) followed by a poly(A)-tail. It encoded a polyprotein of 2227 amino acids (aa) Sequence comparison showed that LU38 shared the highest identities of 98.1% for nt (140 differences) and 99.2% for as (17 differences) with AH1, and the lowest identities of 91.4% for nt (741 differences) with HM175 and 98.1% for aa (43 differences) with GBM. LY6 shared the highest identities of 97.4% for nt (196 differences) and 98.7% for aa (28 differences) with H1 and the lowest identities of 91.2% for nt (642 differences) with HM175 and 97.7% for as (51 differences) with GBM. The subgenotyping revealed that the LU38 and LY6 isolates are of IA subgenotype. The phylogenetic analysis showed that LU38 is closest to AH1 and the LY6 to FH3, suggesting that the epidemiological link of hepatitis A (HA) had developed in China and Japan. PMID- 12580379 TI - Molecular characterization of Hepatitis C virus (HCV) core region in HCV-infected Thai blood donors. AB - In order to investigate the distribution of Hepatitis C virus (HCV) genotypes in Thailand, we performed phylogenetic analysis based on the virus core region and in this way we identified and reliably distinguished HCV genotypes 1-6 as well as subtypes. Among 100 plasma samples randomly selected from blood donors positive for antibodies to HCV (anti-HCV) 90 (90%) were found positive for HCV RNA and 77 of them were subjected to nucleotide sequencing. The following types and subtypes were identified in this group: 1a in 16 samples (20.8%), 1b in 14 samples (18.2%), 3a in 29 samples (37.7%), 3b in 5 samples (6.5%), and 6a in 13 samples (16.9%). Although this study allowed identification and characterization of HCV among blood donors, more extensive studies are needed to explore the HCV distribution in other population groups and in other geographical regions and to exploit the virus core-based characterization of HCV for evaluation of treatment and clinical outcome and epidemiological purposes. PMID- 12580378 TI - Upstream AUGs modulate prion protein translation in vitro. AB - The level of expression of the host PrP gene (PRNP) has been shown to affect the progression to a disease, transmissible spongiform encephalopathy (TSE). In order to define sequences that are responsible for translation ofPRNPmRNA we have investigated a region comprising its 5'-leader sequence. Most remarkable, it consists of an almost identical Kozak mRNA sequence and two AUG initiation codons which seem to modulate translation of the prion protein mRNA in vitro. Although transcriptional regulation of the prion protein PRNP gene had been expected to dominate the translational modulation, our observations point to a translational regulation of the mouse prion protein synthesis controlled by ribosomal entry and usage of AUG codons. PMID- 12580380 TI - Comparison of six Australian isolates of Bovine herpes virus 2 based on UL24 gene after a passage in MDBK cells. AB - A region of the UL24 gene of six Australian field isolates of Bovine herpesvirus 2 (BHV-2) was sequenced after a passage in Madin-Darby bovine kidney (MDBK) cells by polymerase chain reaction (PCR). While the PCR product covered the first half of the UL24 gene, a particular interest was focused on the 274-297 nucleotide (nt) region in which a two nt deletion had previously been detected in the BHM-1 strain of BHV-2. Most isolates tested did not generate any defective UL24 genes during the passage. However, a third of the UL24 genes of BHM-1 strain contained the two nts deletion, but only when a high multiplicity of infection (MOI) was used. Also in the isolate 554 at least a half of the UL24 genes were found to be altered independently of the MOT used. These UL24 genes had an insertion of four nts within the 274-297 nt region. The predicted truncation of the UL24 protein of both viruses occurred at the same stop codon. The region of the gene in which these mutations of the UL24 gene occurred is common to all herpesviruses. PMID- 12580381 TI - Hepatitis C virus phylogeny: a useful clinical tool. AB - In order to type 45 recent isolates of Hepatitis C virus (HCV) originating from four different geographic regions of the world, we performed phylogenetic analysis of a 192 nucleotides (nts) long sequence from the 5'non-coding region (5'-NCR) of the virus genome and compared them with 55 HCV isolates/strains of known type. The results of this study showed that phylogenetic studies can assign an HCV isolate to the correct type in 100% and to the correct subtype in 98%. A comparison ofthis method with other methods using commercial kits revealed that it is appropriate for clinical use and is cost effective. PMID- 12580382 TI - Effect of human cytomegalovirus and glucose on adhesion molecules expression in cultured human endothelial cells. AB - The aim this study was to investigate the effect of glucose on the induction of adhesion molecules by Human cytomegalovirus (HCMV) in endothelial cells in vitro. Primary cultures of human umbilical vein endothelial cells (HUVECs) pretreated with 16.5 mmol/l glucose for 24 hrs were infected with a HCMV strain with tropism for endothelial cells. Expression of adhesion nmolecules (ICAM-1, VCAM-1 and ELAM 1) was measured by flow cytometry. While high concentrations of glucoseperse activated the expression of all three adhesion molecules tested, HCMV induced the expression of ICAM-1 only. Moreover, it potentiated the expression of ICAM-1 in glucose-pretreated HUVECs, while it did not affect at all or slightly suppressed the glucose-activated expression of VCAM-1 and ELAM-1. The modulatory effect of glucose and HCMV on the expression of adhesion molecules in endothelial cells may be applied in increased vulnerability to patients with diabetes mellitus or atherosclerosis. PMID- 12580383 TI - Inhibition of Japanese encephalitis virus maturation and transport in PS cells to cell surface by brefeldin A. AB - A brefeldin A (BFA) treatment of porcine stable kidney (PS) cells resulted in inhibition of Japanese encephalitis virus (JEV) maturation and its transport to the cell surface. Intrestingly, the antigenicity of the virus, in contrast, remained unaffected as no difference in epitope presentation/expression was observed in BFA-treated and control (untreated) infected cells even though in the former cells a loss of hemagglutinating (HA) activity was recorded. Thus it seems that the BFA treatment did not affect the glycoprotein E (gpE) synthesis and folding essentially required for the epitope presentation/expression in cells. PMID- 12580384 TI - Using a population-based health information system to study child health. AB - OBJECTIVE: This paper describes the population-based analyses of measures of child health status used throughout this supplement. METHODS: The articles in this supplement examine health-related data for children 0 to 19 years. Most analyses cover the period from April 1, 1994 to March 31, 1999. Administrative and survey data were used to assess child health and well-being. For regional comparisons, data were broken down by subregions of Manitoba, called Regional Health Authorities (RHAs), and neighbourhoods of Winnipeg, called Winnipeg Community Areas (Winnipeg CAs). The premature mortality rate (PMR) was used as a proxy of the overall health of the population. All graphs comparing rates among RHAs and Winnipeg CAs rank these subregions in the same order, from lowest to highest PMR. Income was operationalized by dividing the province's population into urban and rural quintiles based upon household income. Other aspects of methodology are discussed. RESULTS: Results are presented in the articles that follow this one. CONCLUSION: The relationships between key child health indicators and geographic and socioeconomic factors for Manitoba children are discussed in the articles following this one. PMID- 12580385 TI - Embedding child health within a framework of regional health: population health status and sociodemographic indicators. AB - OBJECTIVE: The description of regional variation in children's health requires regional population-based context. But what is the best way to measure the health of a region's population? METHODS: The use of two indicators is described--one a health status measure and the other a measure of socioeconomic wellbeing. It is well known that the population's premature mortality rate (PMR), the age/sex adjusted rate of death before age 75 years, is highly related to overall health status of an area's residents. Socioeconomic characteristics of an area's residents are also indicative (and likely causative) of health status differences. RESULTS: The Socioeconomic Factor Index (SEFI) was developed at the Manitoba Centre for Health Policy, using a Principal Components Analysis of census data. PMR and SEFI are highly correlated (Spearman's correlation coefficient r = 0.85, p < 0.0001). CONCLUSION: PMR can be used as a surrogate measure for both the health status and socioeconomic well-being of regional populations in Manitoba. PMID- 12580386 TI - A matter of life and death for Manitoba's children: an overview of birth rates and mortality rates. AB - OBJECTIVE: To determine the fertility and child mortality rates for Manitoba. METHODS: Fertility and mortality rates were derived from the Population Health Research Data Repository and Vital Statistics, for 1994 through 1998. Data are presented by 12 Regional Health Authorities (RHAs), 12 Winnipeg Community Areas (CAs) and by income quintile. Each indicator is correlated with PMR (the age- and sex-adjusted premature mortality rate, i.e., death before age 75) and SEFI (Socioeconomic Factor Index, a standardized composite index), both considered proxies for overall health and socioeconomic well-being of populations. RESULTS: Manitoba's total fertility rate was 1.77 children per woman, ranging from 1.62 to 3.15 by RHA, and 1.21 to 2.30 by Winnipeg CA. Manitoba's infant mortality rate was 6.6/1000 (or 5.5/1000 excluding < 500 g or < 20 weeks gestation), ranging from 4.5 to 10.2 by RHA (4.2 to 9.8 exclusive), and 3.7 to 8.4 by Winnipeg CA (2.7 to 6.7). There was a gradient of infant mortality by income quintile (p < 0.001), with double the rate comparing lowest to highest. Child mortality rates varied geographically and by gender, with northern children at greatest risk. Injury was the leading cause of death (52% for ages 1 through 9, 75% for ages 15 to 19). CONCLUSION: Fertility rates, as well as infant and child mortality rates, were positively associated with PMR and SEFI, with substantial geographical variation. PMID- 12580387 TI - Preventable feto-infant mortality: application of a conceptual framework for perinatal health surveillance to Manitoba perinatal outcomes. AB - BACKGROUND: Perinatal health surveillance systems have lacked conceptual frameworks to translate data into information for policy and program planning. This paper demonstrates the application of a conceptual framework in the analysis of feto-infant mortality data in the province of Manitoba. METHODS: Fetal and infant deaths were categorized according to a two-dimensional framework of birthweight and age-at-death, and grouped into four broad categories of contributors to perinatal health: Maternal Health, Maternal Care, Newborn Care, and Infant Care. Birth Weight Proportionate Mortality Rates (BWPMR) were calculated for each of the four categories, and preventable "excess" feto-infant mortality was estimated through comparisons to a benchmark sub-population. RESULTS: Between 1985 and 1998, feto-infant mortality declined from 12.3 to 9.8 deaths per 1000 births in Manitoba. Much of this decline occurred in the Newborn Care category; there were only slight declines in deaths attributed to Maternal Health and Infant Care factors. Comparison of the feto-infant mortality rate to the benchmark rate revealed an excess of 3.46 deaths per 1000 births, an "opportunity gap" of 33%. Substantial regional variations in feto-infant mortality rates were observed. SUMMARY: Application of this conceptual framework provided useful information to aid in policy and program planning. As the greatest excess feto-infant mortality was observed in the Maternal Health and Infant Care categories, attention to the broader determinants of health which influence these categories will be required. PMID- 12580388 TI - Being born in Manitoba: a look at perinatal health issues. AB - OBJECTIVE: The Manitoba Centre for Health Policy was commissioned by Manitoba's provincial health department to examine the health of newborns born 1994 through 1998, using three indicators: preterm birth (< 37 weeks gestation), birthweight, and type of infant feeding. METHODS: Data were derived from the Population Health Research Data Repository and the National Longitudinal Survey of Children and Youth 1996. Variation by 12 Regional Health Authorities (RHAs) and by 12 Winnipeg Community Areas (CAs) was examined, as well as associations with the population's health and socioeconomic well-being. RESULTS: Manitoba's preterm birth rate was 6.7% of live births, from 5.3% to 7.4% by RHA, and 5.7% to 8.0% by Winnipeg CA. Manitoba's low birthweight rate (< 2500 g) was 5.3%, from 2.7% to 5.7% by RHA, and 4.4% to 7.2% by Winnipeg CA. The lower the income, the greater the likelihood of low birthweight (p < 0.05). Manitoba's breastfeeding initiation rate was 78%, from 64% to 87% by RHA, and 66% to 90% by Winnipeg CA. The lower the income and the poorer the health status of the population, the lower the breastfeeding rate (p < 0.001). Of those initiating breastfeeding, 42% breastfed for at least six months. CONCLUSION: Factors affecting child health in Manitoba could be addressed through systematic programs both during pregnancy and during the postpartum period, including support for nutritional counselling, promotion of breastfeeding, smoking cessation programs, and social policy decisions designed to overcome disparities within low-income groups and populations with poorer health status. PMID- 12580389 TI - Factors affecting adolescent reproductive health in Manitoba. AB - OBJECTIVES: To report teen pregnancy and sexually transmitted infections (STI) rates among Manitoba adolescents, and associated factors including rates of sexual intercourse and contraceptive use. METHODS: Teen pregnancy rates in females aged 15 to 19 for the fiscal years 1994/95 through 1998/99 were derived from the Population Health Research Data Repository and reported by geographical areas and income quintiles. Premature mortality rate (PMR) and the Socioeconomic Factor Index (SEFI) measured the overall health and socioeconomic well-being of regional populations. Data on sexual activity and contraceptive use were derived from the 1996 National Population Health Survey for males and females ages 15 through 19 years. RESULTS: The teen pregnancy rate for Manitoba was 63.2/1000, varying by geography and inversely correlated with income, PMR, and SEFI. 39% (95% CI 33-45) of teens reported sexual intercourse, with higher rates in urban areas (46%, 95 % CI 35-57) and the North (48%, 95% CI 36-60) compared to South Rural (30%, 95% CI 25-34), and in low-income families (68%, 95% CI 53-83) compared with middle/high (33%, 95% CI 26-40). For sexually active females, 42% (95% CI 28-57) used the birth control pill, with higher rates in low-income families (70%, 95% CI 50-90) compared to middle/high income (31%, 95% CI 14-48). Condom use (at last sexual intercourse) was reported by 82% (95% CI 72-92) of adolescents, with trends (though not statistically significant) to lower use in low-income families and the North. CONCLUSION: Reliance on the pill for contraception, combined with low rates of condom use, are public health concerns for adolescents where STI and unintended pregnancy rates are high. PMID- 12580390 TI - Assessing health status in Manitoba children: acute and chronic conditions. AB - BACKGROUND: Numerous child health status measures have been developed, ranging from assessments of physical and mental health to activity continuums. Our objective was to report the regional distribution of physical morbidity among children in Manitoba. METHODS: Using Manitoba's population-based prescription and health care data for 1998/99, the prevalence of children with lower respiratory tract infections, four chronic conditions (asthma, cardiovascular disease, Type 1 diabetes mellitus and seizure disorders) and physical disabilities, including spina bifida and cerebral palsy, was determined for 12 Regional Health Authorities and 12 Winnipeg Community Areas, ranked by a measure of population healthiness, the premature mortality rate (PMR). Prescription rates were also reported by neighbourhood income quintile, derived from census data. RESULTS: Hospitalization for lower respiratory tract infection was highest in infants (6%) and increased with successive decreases in neighbourhood income or in the population healthiness of a region. On the basis of a physician diagnosis or prescription drug for asthma, 10% of school-age children had asthma. Asthma treatment rates in northern Manitoba were substantially lower than in Winnipeg. Treatment rates for cardiovascular conditions, Type I diabetes and seizure disorders approached 1% in adolescents and there were no regional differences in the distribution of these conditions. The prevalence of physical disability was highest in northern Manitoba. CONCLUSION: A minority of Manitoba children suffer from chronic and serious acute health problems in childhood, but the burden of illness is not evenly distributed among children. PMID- 12580391 TI - Childhood injury rates in Manitoba: socioeconomic influences. AB - BACKGROUND: Injury is the leading cause of death among Canadian children between 1 and 19 years, and accounts for one sixth of all hospitalizations of children between 0 and 19 years. We examined the causes of injury in Manitoba children, and the relationship between injury rates and region of residence, premature mortality rate (PMR), and income. METHODS: Regional differences in injury death and hospitalization rates, and causes of injury were derived from the Population Health Research Data Repository. The relationship between injury rates and area income levels was assessed and correlations between regional premature mortality rates (PMR) and injury rates were calculated. RESULTS: Motor vehicle crashes were the leading cause of injury mortality. Falls were the leading cause of injury hospitalization. Regional differences were substantial. Rural-urban differences in injury rates were pronounced; northern Manitoba's rates were very high compared to the rest of the province. Regional PMR values correlated significantly with injury mortality and hospitalization rates. Both types of injury rates correlated significantly with income; higher injury rates were associated with lower income levels. CONCLUSION: Injuries are not random events, but are related to social factors. PMID- 12580392 TI - Health service utilization by Manitoba children. AB - OBJECTIVE: To compare physician and hospital utilization rates by children across subregions of Manitoba. METHODS: 1998/99 data for physician visits and hospitalizations for children aged 0 to 19 were extracted from the Population Health Research Data Repository. Rates of utilization were compared across 12 regions (RHAs) within Manitoba, and 12 community areas within Winnipeg. Rates were also compared across premature mortality rates (PMR) and area income levels. RESULTS: Substantial regional variation was found for utilization rates. The hospitalization rate for children from the three northern RHAs (highest PMRs) (114/1000) was almost four times the Winnipeg rate (30/1000) and almost double the rate for the rural south RHAs (lowest PMRs) (59/1000). The variation among regions in physician visits ranged from under 2 visits in 2 of the northern RHAs to almost 4 visits in urban areas. However, the low visit rates in rural RHAs are offset somewhat by greater use of nurses. Hospitalizations and physician visits were also related to area income level. CONCLUSION: Findings are discussed in terms of health care need. PMID- 12580393 TI - Prescription medications in Manitoba children: are there regional differences? AB - BACKGROUND: Population-based studies of pharmaceutical use in children provide information on disease prevalence, physician practice and adherence to treatment. We undertook an evaluation of regional differences in prescription drug use by Manitoba children. METHODS: Using Manitoba's population-based prescription data for 1998/99, the prevalence of children receiving prescriptions for antibiotics, analgesics, iron supplements, and four classes of psychotropic drugs was reported for Regional Health Authorities and Winnipeg Community Areas, ranked by a measure of population healthiness, the premature mortality rate (PMR). Prevalence rates were also reported by census-based neighbourhood income areas. RESULTS: 60% of children received at least one prescription in 1998/99. Antibiotics, antiasthmatics, analgesics, antidepressants, and psychostimulants were the most commonly dispensed drugs. Prescription use of antibiotics, iron supplements, analgesics, antidepressants, antipsychotics and anxiolytics was highest in low income, urban neighbourhoods. Few associations between a region's PMR and prescription utilization were observed, but children living in regions with the least healthy populations were more likely to use antibiotics, non-steroidal anti inflammatory drugs and anxiolytics. Psychostimulant use was unrelated to neighbourhood income, but highest rates were documented in some of the healthiest Winnipeg neighbourhoods. CONCLUSION: We documented regional variation in prescription use which may be related to differences in health, physician practice or child use. PMID- 12580394 TI - Community resources and determinants of the future health of Manitobans. AB - BACKGROUND: Life history studies in health show that some of the key determinants of health inequalities lie in biological and social experiences at the earliest times of life. The objectives of this research were to describe the regional distribution of childhood determinants of adult health, such as school achievement, and the environments which contribute to their development. METHODS: Using Manitoba data from the National Population Health Survey, the National Longitudinal Survey on Children and Youth, the Department of Education, Training and Youth, the Department of Family Services and Housing, the Library Association website and the Agriculture and Food website, the regional distribution of Grade 3 standards test scores and neighbourhood resources such as child care services, libraries, sports participation and food costs were determined for 12 Regional Health Authorities and 12 Winnipeg Community Areas, ranked by a measure of population healthiness, the premature mortality rate. Findings were also reported by income level and larger geographic regions. RESULTS: Children living in neighbourhoods with less healthy populations were more likely to have poorer school performance, as indicated by Grade 3 math standards test scores. They were also more likely to change schools, less likely to participate in sports, and had decreased access to affordable food and licenced day care. They had similar access to library books as children living in more healthy neighbourhoods, although book lending rates were not measured. CONCLUSION: We documented regional variation in the availability of resources to support healthy childhood development. PMID- 12580395 TI - The virtual classroom: a summary of child health indicators. AB - OBJECTIVE: To provide an overview of child health indicators and health care utilization patterns in Manitoba by comparing child health outcomes for different income groups: a) children from two different community areas of Winnipeg (Fort Garry and Point Douglas), and b) adolescents from two different parts of Manitoba (the North, and Winnipeg). METHODS: Various child health indicators derived from population-based administrative data and national surveys are reported in the articles within this supplement. Childhood morbidity and mortality, health care utilization patterns, pharmaceutical use, and regional demographic information discussed in the research articles in this supplement are summarized here using comparisons of outcomes in "virtual classrooms" of 100 children. RESULTS: Large gradients were observed in the comparison of the virtual classrooms of 100 high school students, including the following numbers of adolescents: females on birth control pills (Winnipeg 11, North 18); injury hospitalization annually (Winnipeg 1; North 4). Gradients are observed for some child outcomes of the virtual classroom of 100, but not for others. Examples include the following numbers of children: preterm at birth (FG 7, PD 7); breastfed at birth (FG 90, PD 66); hospitalized for lower respiratory tract infection in first year (FG 3, PD 8); parents having no high school (FG 11; PD 41). CONCLUSION: Throughout Manitoba, child and adolescent health indicators and determinants of health show gradients by income as well as by geographical regions. PMID- 12580396 TI - The role of tumor oxygenation in vascular and clinical response to angiogenesis inhibition. PMID- 12580397 TI - Comparison of fluctuations of oxygen tension in FSA, 9L, and R3230AC tumors in rats. PMID- 12580398 TI - Effect of carbogen-breathing on redox status of the RIF-1 tumor. PMID- 12580399 TI - Evaluation of microregional variations in tumor hypoxia following the administration of endostatin. PMID- 12580400 TI - Effect of hydralazine on blood flow, oxygenation, and interstitial fluid pressure in subcutaneous tumors. PMID- 12580401 TI - Oxygenation of cervix cancers: impact of clinical and pathological parameters. PMID- 12580402 TI - Quantitative spatial analysis of hypoxia and vascular perfusion in tumor sections. PMID- 12580403 TI - Tumor oxygenation and its relevance to tumor physiology and treatment. PMID- 12580404 TI - Mechanisms underlying hypoxia development in tumors. PMID- 12580405 TI - Lack of association between tumor hypoxia, GLUT-1 expression and glucose uptake in experimental sarcomas. PMID- 12580406 TI - The impact of tissue microenvironment on treatment simulation. PMID- 12580407 TI - Evidence for and against hypoxia as the primary cause of tumor aggressiveness. PMID- 12580408 TI - Characterization of human Gc protein-derived macrophage activation factor (GcMAF) and its functional role in macrophage tumoricidal activity. PMID- 12580409 TI - Effect of hemoglobin solutions as hemodiluents on capillary oxygen tension. PMID- 12580410 TI - Different efficacy in vitro of hemoglobin based oxygen carriers and red cells. PMID- 12580411 TI - A novel blood transfusant candidate: intact human erythrocytes containing hemoglobin exclusively nitrosylated in the alpha-subunits. PMID- 12580412 TI - The contribution of hemoglobin to oxygen transport in the microcrustacean Daphnia magna. A conceptual approach. PMID- 12580413 TI - Modifications of red blood cell shape and glycoproteins membrane content in septic patients. PMID- 12580414 TI - Fiber optic immunosensors for cardiovascular disease diagnosis: quantification of Protein C, Factor V Leiden, and cardiac Troponin T in plasma. PMID- 12580415 TI - Soft gel chromatography column analysis and design for the production of high molecular weight blood factors: model molecule Protein C. PMID- 12580416 TI - Development of an economic miniantibody production process for the purification of Protein C (anti-coagulant/anti-thrombotic). PMID- 12580417 TI - Nitric oxide synthesis in brain is stimulated by oxygen. PMID- 12580418 TI - Oxygen indirectly regulates nitric oxide availability. PMID- 12580419 TI - Modeling the regulation of oxygen consumption by nitric oxide. PMID- 12580420 TI - Oxygen sensor design: analysis and correction of problems associated with zero current, stability and structure. AB - Membrane-covered polarographic sensors were designed by Leland Clark several decades ago. The membrane covering was a brilliant innovation because it provided an isolation of electrochemical elements from the external environment. These sensors have found a wide range of applications in many aspects of medicine, research and industry but they do not perform well at low oxygen levels because of high and unpredictable zero-current and drift characteristics. The key to overall improved zero-current and stability was a highly improved cathode seal design. Rather than heat-sealing wire into glass microtubing, a high temperature molding procedure was employed. Other improvements involved substantial modifications to the conventional methods of fixing the membrane in place, eliminating all crevices at the periphery of the membrane. This guards against potential contamination, entrapment of gas bubbles and carry over. The present sensor design can operate in gases and liquids with zero current less than 0.02% oxygen. PMID- 12580421 TI - Computer simulation of oxygen microelectrode measurements in tissues. PMID- 12580422 TI - Physiological oxygen measurements using oxygen electrodes. PMID- 12580423 TI - Accuracy of a polarographic electrode at high oxygen concentrations. PMID- 12580424 TI - Recessed oxygen electrodes: getting more than PO2. PMID- 12580425 TI - Oxygen distributions in tissue measured by phosphorescence quenching. PMID- 12580426 TI - Adapting and validating a NIRS device to measure in utero fetal sheep cerebral oxygenation. AB - NIRS has provided us with a reliable and sensitive method to monitor in utero fetal cerebral oxygenation in the surgically-recovered pregnant sheep. Our preliminary results indicate that monitoring fetal cerebral oxygenation may be important to understanding how maternal drug exposure can affect the fetal brain. PMID- 12580427 TI - Detection of the fast neuronal signal on the motor cortex using functional frequency domain near infrared spectroscopy. AB - Using non-invasive near infrared spectroscopy fast changes in the range of ms in the optical properties of neurons during brain activity have been described. Since the signal is small, the system to detect it has to be highly noise optimized. We used a frequency-domain tissue oximeter, whose laser diodes were modulated at 110 MHz and the amplitude (AC), mean intensity (DC) and phase (phi) of the modulated optical signal was measured at 96 Hz sample rate. In two volunteers, 36 and 37 years old, the probe consisting of 4 crossed source detector pairs was placed above the motor cortex (C3 position), contralateral to the hand performing the tapping exercise. The tapping frequency was set at 2.5 times the heart rate of the subject to avoid the influence of harmonics on the signal. An electronic device recorded the tapping movement. Control-data were obtained from a solid medium of approximately the same optical properties as the human head. To reduce physiological noise the arterial pulsatility was removed using an adaptive filter, the data was detrended by a high pass filter and a cross correlation function between the optical data and the tapping signal was calculated. The instrumental noise of the control data was very low (AC mean 0.0015% +/- SD 0.00092%, DC 0.00037% +/- 0.00023% and phi 0.00083 degrees +/- 0.00042 degrees). On the head the noise level was AC 0.0042% +/- 0.0031%, DC 0.0021% +/- 0.0012% and phi 0.0020 degrees +/- 0.0017 degrees. In 14 DC, 5 AC and 0 phi out of 30 locations a fast signal was detected, which was higher (p<0.001) than the noise level. This signal disappeared during non-tapping periods. With the signal to noise ratio that we have achieved single subject measurements become feasible. PMID- 12580428 TI - Probing rat brain oxygenation with near-infrared spectroscopy (NIRS) and magnetic resonance imaging (MRI). PMID- 12580429 TI - Variation in blood oxygenation and cerebral pO2 in a mouse model measured by EPR spectrometry. PMID- 12580430 TI - Functional oxygen-17 magnetic resonance imaging and localized spectroscopy. AB - Functional oxygen-17 magnetic resonance (MR) imaging and localized spectroscopy is defined as the ensemble of MR measurements aiming at in vivo, noninvasive characterization of oxygen transport and utilization. After a brief description of the present status of in vivo 17O-MR, preliminary results are reported on oxygen delivery and consumption in cell suspensions of Saccharomyces cerevisiae. It is shown that parallel 31P-MR at high magnetic fields has an important corroborative value. PMID- 12580431 TI - Development of a retractor for monitoring the oxygenation state of brain tissue. PMID- 12580432 TI - Mapping of hemodynamics on the human calf with near infrared spectroscopy and the influence of the adipose tissue thickness. AB - We investigated the influence of the adipose tissue thickness (ATT) on near infrared spectroscopy (NIRS) measurements of the absorption coefficient (mu a), the reduced scattering coefficient (mu s') and changes in concentrations of oxyhemoglobin ([O2Hb]) and deoxyhemoglobin ([HHb]). We used a frequency domain spectrometer and a special probe to generate maps of these parameters on the human calf during venous occlusion. For ATT below 6 mm mu a remained constant, whereas for ATT between 6 and 14 mm mu a decreased quickly and became almost constant again for ATT larger than 14 mm. Mu s' was not significantly altered by the ATT but the values showed a high variability between subjects. We found significantly different changes in both the [O2Hb] and the [HHb] between the proximal and distal locations of measurement. Although ATT influences the recovery of the optical properties of the underlying tissue, these differences depending on the location cannot be sufficiently explained by the ATT for the following reasons. The ATT varied little within one subject (mean difference 0.88 +/- 1.80 mm). The inter-subject variability was 5 times higher. For a given ATT within one subject we observed different values for changes in [O2Hb] and [HHb] depending on the measurement location. Moreover for a smaller ATT the difference between the values of delta[O2Hb] and delta[HHb] proximal versus distal were more distinct. The thinner the overlying tissue (ATT) the higher is the proportion of muscle tissue in the probed tissue volume. Therefore these differences are most likely coming from the muscle tissue rather than the ATT. This indicates that although the ATT has an evident influence on the measurement of optical parameters and hemodynamics and should therefore be recognized when performing NIRS measurements, other factors will have to be considered as well. PMID- 12580433 TI - EPR imaging of vascular changes in oxygen in response to carbogen breathing. PMID- 12580434 TI - Molecular imaging by PET. PMID- 12580435 TI - Frequency domain imaging of oxygen tension in the mouse retina. Preliminary instrumentation development. PMID- 12580436 TI - Oxygen delivery to the retina and related visual pathology. Brief review. PMID- 12580438 TI - Effect of blood flow on PvO2-VO2 relation in contracting in situ skeletal muscle. PMID- 12580437 TI - Activation of the visual cortex in newborn infants under natural sleep using multichannel near-infrared spectroscopy. PMID- 12580439 TI - Oxygen transport and intracellular bioenergetics on stimulated cat skeletal muscle. AB - A unique multiparameter recording of skeletal muscle bioenergetics, biochemistry and biomechanics has permitted determination of novel relationships among hemodynamics, cellular high-energy metabolites and mitochondrial bioenergetics in feline skeletal muscle. The study utilizes 31P NMR, NIR, and NADH fluorescence spectrophotometry, biochemical assays and muscle performance. Seven cats were anesthetized and mechanically ventilated. Calf muscles were stimulated through sciatic nerve electrical stimulation and tension was monitored by a strain gauge connected to the Achilles tendon. We stimulated the muscle to produce several workloads up to Vmax. We also changed FiO2 from normoxia to hypoxia for each %Vmax. From these results, the most sensitive indicators of cellular hypoxia leading to a reduction in muscle performance can be determined. Hemoglobin deoxygenation generally does not correlate with cellular hypoxia, although when the HbO2 drops below 30% saturation there is an increased incidence of cellular hypoxia. The [ADP], which is known to regulate mitochondrial function, has a close relation to the work, not to the hypoxia. On the other hand, the mitochondrial NADH does respond to cellular PO2. The degree of oxidation (NADH decrease) due to the ATP flux shifts with oxygen availability in mild to moderate hypoxia (at FiO2 down to 9%). As cellular hypoxia causes decreases in muscle performance (moderate to severe hypoxia), NADH is being reduced rather than oxidized with increasing workloads. PMID- 12580440 TI - Muscle oxygenation by near infrared spectroscopy and lactate thresholds in endurance trained and recreationally active cyclists. PMID- 12580441 TI - Cardiac output, oxygen consumption and muscle oxygen delivery in submaximal exercise. Normal and low O2 states. AB - Cardiac output (Q) changes linearly with oxygen consumption (VO2) in normal subjects undertaking submaximal exercise (Q = A + B x VO2 where A is the y intercept and B the slope). If (hypothesis 1) the increase in cardiac output above the resting state represents the blood flow to exercising muscle (qm) and the increase in VO2 represents the oxygen consumption of exercising muscle (VO2m) then, where CaO2 is the arterial oxygen content, oxygen extraction, Em = 1/(B x CaO2). Secondly, exercising muscle venous oxygen content, CvO2m = CaO2 - 1/B. Limiting the hypothesis just to the calculation of VO2m (hypothesis 2) allows calculation of qm if CaO2 and CvO2m are available. From Koskolov et al. (Am. J. Physiol.: Heart and Circ. Physiol. 273, H1787-H1793, 1997), exercising muscle blood flow (qm) is equal to the increment in cardiac output when CaO2 is normal but exceeds it when CaO2 is low. Muscle Oxygen extraction (Em) is found to be 68% in submaximal exercise. Hence, muscle oxygen delivery (DaO2m) for a given metabolic rate is sustained in low O2 states (at 1.48 ml DaO2m per ml VO2m), confirmed by analysis of Roach et al. (Am. J. Physiol.: Heart and Circ. Physiol. 276, H438-H445, 1999). PMID- 12580443 TI - Diffuse optical measurement of hemoglobin and cerebral blood flow in rat brain during hypercapnia, hypoxia and cardiac arrest. AB - We have demonstrated the ability to concurrently measure relative changes in cerebral blood flow, hemoglobin concentration, and hemoglobin oxygenation with a single non-contact, non-invasive instrument. Our measurements from rat hypercapnia, hypoxia and cardiac arrest models are in reasonable agreement with the literature, and offer the possibility for further growth and quantification. The optical techniques used in this study are attractive also because they enable experimenters to measure vascular response of deep tissues. The new instrument and concept may also be applicable to human studies especially in infants and neonates permitting noninvasive monitoring of cerebral hemodynamics and oxygen (see [4] and [2] for examples of NIR spectroscopy). PMID- 12580442 TI - Non-invasive PET and SPECT imaging of tissue hypoxia using isotopically labeled 2 nitroimidazoles. AB - The measurement of pathologically low levels of tissue pO2 is an important diagnostic goal for determining the prognosis of many clinically important diseases including cardiovascular insufficiency, stroke and cancer. A class of bioreductively activated drugs, typified by the 2-nitroimidazoles, has excellent potential for application to this goal. Such drugs bind to cells at a rate which is maximal under conditions of severe hypoxia (e.g. less than 0.05% oxygen) and is inhibited, with Michaelis-Menten kinetics, as a function of increasing oxygen concentration. A number of detection possibilities exist for the drug adducts, including invasive assays which can measure drug adducts in tissue sections at cell-to-cell resolution. Use of such agents in non-invasive assays is important and, to this end, a number of drugs have been conjugated with radioactive isotopes suitable for detection by Nuclear Medicine techniques. In contrast with the invasive assays, resolution and contrast is much more limited with the non invasive assays. Thus, there are many factors contributing to the balance of pros and cons for the non-invasive vs. invasive use of 2-nitroimidazole drugs as hypoxia detectors. These factors will be summarized in this review, with emphasis on compounds suitable for clinical use. PET (positron emission tomography) imaging with 18F-labeled EF5 (a drug in current clinical trials using invasive assays) will be described. PMID- 12580444 TI - Multiorgan monitoring of hemodynamic and mitochondrial responses to anoxia and cardiac arrest in the rat. PMID- 12580445 TI - Importance of glomus cell plasma membrane and mitochondrial membrane potentials during acute hypoxia signaling in the rat carotid body. PMID- 12580446 TI - Estimation of hydroxyl radicals based on the salicylate trapping method in hippocampus of stroke-prone spontaneously hypertensive rats (SHRSP) during transient ischemia and recirculation. PMID- 12580447 TI - Effect of hypoxia and ischemia on expression of selected genes in brain of newborn piglets. PMID- 12580448 TI - Brain oxygenation during cardiopulmonary bypass and circulatory arrest. AB - Quantitative measurements of oxygen distribution in the microcirculation of the brain cortex of newborn piglets were made during different modes of cardiopulmonary bypass. Three groups of animals, anesthetized and mechanically ventilated, were studied. The first group of animals were maintained on normothermic cardiopulmonary bypass (CPB) at a flow of 100 ml/kg/min, while the second and third groups underwent low flow hypothermic cardiopulmonary bypass (40 ml/kg/min at 18 degrees C) (LFCPB) and deep hypothermic (18 degrees C) circulatory arrest (DHCA), respectively. After bypass, the piglets were monitored for a two hours post-bypass recovery period. CPB caused a decrease in the cortical oxygen from 62 +/- 3 mm Hg to 32 +/- 7 mm Hg at the beginning of bypass and to 36 +/- 5 mm Hg at the end of bypass. During the recovery period, cortical oxygenation steadily decreased, reaching 29 +/- 8 mm Hg at the end of the experiment. With initiation of LFCPB, cortical oxygen decreased to 22 +/- 7 mm Hg. Upon rewarming cortical oxygen increased to 37 +/- 5 mm Hg and then decreased again to about 30 mm Hg at the end of two hours of post-bypass recovery. Similar changes in cortical oxygenation were observed during DHCA. In DHCA cortical oxygen decreased to 19 +/- 4 mm Hg and during rewarming and recovery increased to 35 +/- 6 mm Hg. In conclusion, it has been shown that in newborn piglets recovering from CPB, LFCPB and DHCA, when the blood pressure remained above 55 mm Hg and therefore total blood flow should be well maintained, oxygen pressure in the microvasculature is significantly lower than for pre-bypass. It is suggested that the decreased oxygenation is due to increased heterogeneity in resistance in the microcirculatory units, resulting in broadened distribution of flow rates and oxygen levels. PMID- 12580449 TI - Expression of angiopoietin-1 and -2 in the rat brain during chronic hypoxia and de-adaptation. PMID- 12580450 TI - Hypoxia-inducible factor-1alpha accumulation in the rat brain in response to hypoxia and ischemia is attenuated during aging. PMID- 12580451 TI - Lung ischemia: endothelial cell signaling by reactive oxygen species. A progress report. AB - These studies using both intact lung and reconstituted cell systems have shown that pulmonary endothelial cells respond rapidly (within several seconds) to the acute cessation of perfusate flow, i.e., ischemia. These effects represent a response to the loss of shear stress and are unrelated to changes in cellular oxygenation. The immediate response is partial depolarization of the endothelial cell membrane followed by activation of endothelial NADPH oxidase and the extracellular generation of superoxide anion. Dismutation of superoxide to H2O2 generates a cell signaling molecule that results in the activation of protein kinases and transcription factors which in turn lead to NO generation and activation of endothelial cell division. The presumed physiological role of this signal cascade is the generation of a vasodilator (NO) and the formation of new capillaries in the effort to restore blood flow. PMID- 12580452 TI - Near infrared spectroscopy (NIRS) measurements during global cerebral ischemia in sheep. PMID- 12580453 TI - Simulations of capillary network oxygen transport during transient ischemia in the presence and absence of tissue myoglobin. PMID- 12580454 TI - Cerebral oxygenation during chronic obstructive pulmonary disease. PMID- 12580455 TI - Physiology of functional activation. PMID- 12580456 TI - Nonlinear analysis of heart rate variability in infants with apparent life threatening events. AB - Some studies have shown that neonates having experienced an Apparent Life Threatening Event (ALTE) have an increased risk of dying of the Sudden Infant Death Syndrome (SIDS). In this study the heart rate variability in neonates has been analyzed through a study of the long range correlations and scale invariance (self-similarity) in their cardiac rhythm. The goal is to see whether it is possible to distinguish normal infants from infants with ALTEs using the previously mentioned nonlinear measures of the heart rate variability. Twelve ALTE infants are included in this study. To determine the degree to which these neonates suffer from ALTEs two examinations are performed: a polysomnography and home monitoring. Using the nonlinear measures obtained from the nonlinear analysis of the heart rate variability, the group suffering most from ALTEs can clearly be separated from the other neonates. PMID- 12580457 TI - The effect of nicotinamide & hyperoxic gases on blood glucose. PMID- 12580458 TI - Measurement of tissue oxygenation index during the first three days in premature born infants. AB - No normal values of tissue oxygenation index (TOI) of the brain are known regarding premature born infants. We measured TOI, a measure for the cerebral hemoglobin oxygen saturation, on the head of 15 preterm infants with a median postmenstrual age of 28 weeks (interquartile range (IQR) between 26-29 weeks) with spatially resolved spectroscopy (NIRO 300, Hamamatsu) during the first three days of life. Infants with intra-ventricular hemorrhage or periventricular leucomalacia before the first measurement, as shown by ultrasound, were excluded. The first measurement was done within the first 6 hours of life, the second and third measurement at, respectively, 24 and 48 hours after this first measurement. The mean TOI was calculated if saturation did not change by more than 5% for at least 30 minutes. Other parameters measured were PaO2, PCO2, pH, mean arterial blood pressure, heart rate, hemoglobin, glycemia and peripheral oxygen saturation. There was a significant increase of TOI after 24 (p < 0.05) and 48 (p < 0.001) hours. The median TOI on the first day was 57% (95% CI: 54-65.7), 66.1% on the second day (95%CI: 61.9-82.3%) and 76.1% on the third day (95%CI 67.8 80.1%). No correlation was found between TOI and peripheral oxygen saturation, blood pressure, PaO2, PaCO2 and hemoglobin concentration after multiple regression analysis. TOI increases in the first three days in premature born babies. The increase of TOI is not due to an increase of oxygenation or mean arterial blood pressure. In our opinion, it reflects the increase in cerebral blood flow during the first three days. PMID- 12580459 TI - Simultaneous measurement of cerebral tissue oxygenation over the adult frontal and motor cortex during rest and functional activation. PMID- 12580460 TI - Dysfunctional microvascular conducted response in irradiated normal tissue. PMID- 12580461 TI - Oxygenation and blood concentration changes in human subject prefrontal activation by anagram solutions. AB - Continued studies in cognitive function in problem solving of graded difficulties has now proceeded through several thousand tests of high school students in a six week period last summer (2000). The optical imager used 9 sources and 23 detectors operated in time multiplex to give an image of the prefrontal region in several seconds. Thus, the image can be acquired in the state prior to activation, during and following activation. The images are acquired at two wavelengths, 750 and 850 nm, from which the changes of blood concentration (blood pooling) and relative oxygenation/deoxygenation state (metabolic activity) can be obtained and with suitable calibrations converted into micromolar changes of total Hb concentration and fractional oxygenation, respectively. The imager has a wearable pad applied to the forehead and the electronics now presents the running average of the two quantities mentioned above in the 16 voxels which cover the prefrontal region in 1 sec. Thus the protocol involves a scanning through anagrams of graded difficulty which are known to include the range of maximal activation (>3 solutions in the 30 sec test interval or are too easy (>10), or too difficult. Anagrams are presented in a sequence of three, four, five, etc. letter anagrams to the maximal level of difficulty and then back down again to the starting point. Each anagram is displayed for approximately one minute. Computer scanning of the results gives histograms of the several hundred tests per individual in the "training" interval (for three weeks) and in the post training interval about 400 tests, each for 4 levels of difficulty (3 weeks). Usually two or three complete trials per day were achieved with the result that the group of 7 students produced over 2600 test results in 2000. The histogram displays are under study to determine: a) the most fruitful voxels, b) the fraction of the total tests that appeared in those voxels, and c) the maximum signal level observed in the histogram display in units of micromolar hemoglobin. The product of these two was taken as the Figure of Merit and their displays of the 16 voxels gave patterns for trained and untrained responses. The preliminary conclusions of this study were: a) that the training effect was very large, pretraining exhibited a chaotic voxel distribution for all difficulty levels while trained students gave a higher output and activated only one or two of the 16 voxels. If the tests were too easy, too hard, i.e., an ability/difficulty mismatch, frustration or disattention gave similar chaotic patterns (7); b) a match between difficulty and ability activated only a few voxels in similar locations. Since attention and success measures appear to be of interest in school room studies, a wearable imager with local LCD display can be supplied to the Senior class of the local high school for those individuals who have already had last Summer's training in anagrams. A wearable NIR measurement of metabolic activation and blood flow may be a useful educational aid. In summary, the optical method focuses attention on cognition activated oxygenation of prefrontal cortex. PMID- 12580462 TI - Quantitation of the concordance between cerebral intravascular oxygenation and mean arterial blood pressure for the detection of impaired autoregulation. AB - Since some important forms of brain injury in premature infants are caused in considerable part by disturbances in cerebral blood flow (CBF), it is important to be able to detect whether the cerebrovascular autoregulation, the mechanism by which CBF is maintained constant despite alterations in mean arterial blood pressure (MAP), is working properly. A recent study suggested that concordant changes in MAP and cerebral intravascular oxygenation (HbD), measured non invasively by near-infrared spectroscopy (NIRS) as the difference between the concentration changes of oxygenated hemoglobin (HbO2) and deoxygenated hemoglobin (Hb), reflect impaired cerebrovascular autoregulation. Consequently, premature infants with impaired cerebrovascular autoregulation could be identified by simultaneous, continuous measurements of HbD and MAP. From several premature babies, MAP, HbD and arterial oxygen saturation (SaO2) were measured simultaneously at the University Hospital Leuven, Belgium. The concordance between MAP and HbD was quantitated using three different measures, among which a newly developed measure that looks for similarity of the dynamics between signals. Some preliminary results obtained from the measured data are given. PMID- 12580463 TI - Effects of fluid percussion injury on rat brain hemodynamics, ionic, electrical activity, and energy metabolism in vivo. PMID- 12580464 TI - Changes in the basic experimental parameters of capillary gas chromatography in the 20th century. AB - Studies of qualitative changes in capillary gas chromatography are of significant practical and scientific interest. This paper analyzes the evolution of the most important experimental chromatographic parameters over the last three decades and is based on the use of a new approach to scientometrical research that is referred to as applied scientometry. One essential feature of this approach is that it looks at the entire contents of each paper rather than only taking account its title, abstract. and references (as is typical for conventional scientometry). In this paper, we monitor how the most important chromatographic parameters, such as column length and diameter, layer thickness, stationary liquid phases, separation temperature mode. etc., have been evolving over the period 1970-2000. We used data from the following journals: Chromatographia, Journal of Chromatography, and Journal of High Resolution Chromatography and Chromatography Communications. PMID- 12580465 TI - Minimum in the temperature dependence of the Kovats retention indices of nitroalkanes and alkanenitriles on an apolar phase. AB - The Kovats retention indices (I) of 1-nitroalkanes and alkanenitriles were determined on polydimethylsiloxane and Innowax (polyethylene glycol) columns in a wide temperature range. The temperature dependence of the retention indices exhibits a definite minimum for the early members of the homologous series. The position of the minimum shifts to lower temperatures with increasing carbon atom number of the solute. The thermodynamic explanation of an extreme in the I vs. T function is the higher solvation heat capacities of nitroalkanes and alkanenitriles relative to those of the reference n-alkanes, owing to the deviation from the ideal state in the solution. A novel equation was derived which describes the minimum in the I vs. T function, too. PMID- 12580466 TI - Quantitative accuracy in the gas chromatographic analysis of solvent mixtures. AB - Quantitative accuracy is of great importance in the analysis of bulk mixtures of solvents, particularly when the analysis is related to quality control of very large product volumes like in solvent recovery plants. Serious errors can be made if the effects of density differences between the pure solvents and volume contractions are not properly addressed. In earlier work, the use of an iterative process for correcting such errors has been suggested. However, in the case of volume contractions and mixtures of several solvents, this procedure is difficult to apply. In the present paper, we describe a simple procedure where calibration curves based on mass concentration are utilized. The densities of calibration mixtures of known compositions are determined with a density meter, in order to provide for correction factors caused by volume contractions. Model experiments with mixtures of water, ethanol, acetone and methanol showed a significant improvement in quantitative accuracy. when the suggested calibration strategy was applied. PMID- 12580467 TI - Comprehensive two-dimensional gas chromatography: metrics, potentials, limits. AB - Metrics for evaluation of separation performance of comprehensive two-dimensional gas chromatography (GCxGC) and for quantitative comparison of that performance with similar performance of its 1D (one-dimensional) counterparts are described. The performance improvement can be expressed via reduction in the saturation of a chromatogram or-in the case of the uniform distribution of peaks along the second dimension--via the peak capacity gain due to GCxGC. An order of magnitude peak capacity gain due to the comprehensive GCxGC is possible under optimal conditions. Optimal parameters of the second dimension column as well as the optimal operational conditions for that column and for the modulator in a comprehensive GCxGC are also presented. PMID- 12580468 TI - Determination of siloxane-water partition coefficients by capillary extraction high-resolution gas chromatography study of aromatic solvents. AB - Partition coefficients of benzene, toluene, ethylbenzene and xylenes (BTEX), between crosslinked polydimethylsiloxane and water, were determined at room temperature by capillary extraction (a form of in-tube solid-phase microextraction, SPME) coupled to open tubular gas chromatography (in-tube SPME high-resolution GC). A series of 7-9 repetitive extractions, performed on a 1-ml volume of diluted aqueous BTEX sample by the double-syringe squeeze method, gave exponential regression curves which fit very well with those predicted by partition theory. From the equations of the curves of relative FID response vs. extraction number, experimental Kd were easily calculated and the results compared with literature values. The whole measurement requires about 1 h from the start of the experiment to the final calculation of all BTEX partition coefficients. In-tube SPME resulted in a fast, clean, efficient, and cheaper alternative than the classic 1-cm, externally coated, SPME fiber-holder technique. PMID- 12580469 TI - Image background removal in comprehensive two-dimensional gas chromatography. AB - This paper describes a new technique for removing the background level from digital images produced in comprehensive two-dimensional gas chromatography (GCxGC). Background removal is an important first step in the larger problem of quantitative analysis. The approach estimates the background level across the chromatographic image based on structural and statistical properties of GCxGC data. Then, the background level is subtracted from the image, producing a chromatogram in which the peaks rise above a near-zero mean background. After the background level is removed, further analysis is required to determine the quantitative relationship between the peaks and chemicals in the sample. The algorithm is demonstrated experimentally to be effective at determining and removing the background level from GCxGC images. The algorithm has several parametric controls and is incorporated into an interactive program with graphical interface for rapid and accurate detection of GCxGC peaks. PMID- 12580470 TI - Carrier gas as a new factor influencing the selectivity of the gas-stationary liquid phase chromatographic system. AB - This paper generalizes studies on the influence of carrier gas on relative and absolute retention values. This line of research is also of importance due to the fact that, in the opinion of many chromatographers, the role of the carrier gas is limited only to transporting analyzed compounds along the column. However, even under conditions of the conventional capillary gas-liquid chromatography (i.e. at column pressures under 5 atm) carrier gas (its nature and pressure) significantly influences retention and separation of the analyzed compounds. First, carrier gas (N2 and CO2, for example) dramatically affects relative retention values. For this reason, one should use limit values of alpha(ij) (0) = lim alpha(ij)(P(av)) and I(i)(0) = lim Ii(Pav) I(0) = limI(i) (Pav) with Pav-->0 as chromatographic constants, rather than traditional relative retention values alpha(ij)(P(av)) and I(i)(P(av)). Second, the average pressure Pav of the carrier gas in a column and the nature of the carrier gas influence the selectivity of the gas-stationary liquid phase chromatographic system. Third, wishing to maximize the role of the carrier gas as a factor that improves separation of analyzed compounds, we should design a special gas chromatograph that would allow work with pressures in the column up to 30-50 atm. PMID- 12580471 TI - Needle-type concentrator and its application to the determination of pollutants. AB - A new method for impurity concentration in gas and liquid media using a needle type microconcentrator was developed. The main advantage of this method using microconcentrators is its efficiency and simplicity. Our work was stimulated by Pawliszyn's solid-phase microconcentration method. PMID- 12580472 TI - New high-performance cryofocalizer injector for in-tube solid-phase microextraction and headspace capillary gas chromatographic applications. AB - The construction of a high-efficiency but cheap injector for volatile and very volatile compounds is shown. The device focuses the compounds in a fused-silica (FS) transfer capillary with the aid of liquid nitrogen. A 6.2 mm O.D. glass tube liner (ca. 25 cm x 1.5 mm I.D.) is inserted in the heated (-200 degrees C) injector of the gas chromatograph in place of the standard glass liner, and extends further externally through a liquid nitrogen container made with styrofoam-like material. Inside this glass tube, the FS transfer line passing through the oven door is connected like a pre-column to the analytical high resolution GC column. It can move fast between the heated and the cooled zone (<- >, deltaL = 13 cm), and when this movement starts, crvofocused analytes are injected "at once" resulting in symmetrical and sharp injection bands with "zero" carryover. The performance of this device is demonstrated by its application to in-tube solid-phase microextraction and to spice volatiles analysis. PMID- 12580473 TI - Symmetrical (double-speed) static coating for high-resolution gas chromatography column preparation. AB - "Breakthrough" during static coating could be avoided using a new coating strategy: while filling the capillary support with coating solution, a short mercury plug was placed at the middle of the support length. Mercury was frozen by a cold source and vacuum was applied to both open ends. In this way static coating became symmetric and so evaporation rate was doubled, on a time base, and more than doubled on a coated length base. Coating time at room temperature varied from approximately 20 min for 3-m columns to approximately 3 h for 15-m columns. PMID- 12580474 TI - Capillary extractors for "negligible depletion" sampling of benzene, toluene, ethylbenzene and xylenes by in-tube solid-phase microextraction. AB - The suitability of "capillary extractors" is demonstrated for the "negligible depletion" extraction of benzene, toluene, ethylbenzene and xylenes in a clean water matrix. Extraction set-up and major extractor parameters (length, internal diameter, and film thickness) are chosen to allow rugged analysis by GC with flame ionization detection. With the selected negligible extraction conditions, the efficiency for every consecutive extraction is about 2-3% of the dissolved amount. PMID- 12580475 TI - In-tube solid-phase microextraction sampler for long-term storage. AB - Capillary extractors are proposed as samplers-preconcentrators to overcome losses of volatile organic compounds found using "classical" solid-phase microextraction fiber-holder samplers. A set of equal-size extractors was used to extract in-tube an aqueous solution of benzene, toluene, ethylbenzene and xylenes (BTEX) at 146 ppb. After storage for 6-30 days at 0-4 degrees C (or -15 degrees C) GC analyses were carried out to study BTEX recovery. Results demonstrated that sample preservation was very good: recovery was higher than 98% after 6 days, and more than 95% after 30 days. Capillary extractors, due to their high performance in preserving sample integrity, represent a real breakthrough for on-site sampling of volatile compounds by solid-phase pre-concentration techniques. PMID- 12580476 TI - Semi-automated hollow-fibre membrane extraction, a novel enrichment technique for the determination of biologically active compounds in water samples. AB - An automated hollow fibre membrane extraction technique was developed for the GC MS determination of pharmaceutical and endocrine disrupting compounds in water samples. Enrichment was carried out inside a porous polypropylene hollow fibre membrane, which separated the aqueous and organic phases and regulated the transfer of analytes. n-Octanol placed inside the hollow fibre was used as the acceptor solution. A water-solvent ratio of about 300:1 was used to concentrate the analytes. After 1 hour's extraction of the water sample under magnetic stirring, 1 microl of the n-octanol phase was automatically injected from the hollow fibre into the GC-MS. Development work included examining the influence of different sample matrices, volumes, extraction times and extraction solvents. The detection limits, linearity and standard deviations of the method were determined using drugs such as ibuprofen, phenazone and carbamazepine as well as the endocrine disrupting compounds. technical nonylphenols, bisphenol A, 17alpha ethinylestradiol and tonalide by way of example. PMID- 12580477 TI - Automated hollow fiber-protected dynamic liquid-phase microextraction of pesticides for gas chromatography-mass spectrometric analysis. AB - Dynamic liquid-phase microextraction (LPME) controlled by a programmable syringe pump was evaluated for extracting pesticides in water prior to GC-MS analysis. A conventional microsyringe with a 1.3-cm length of hollow fiber attached to its needle was connected to a syringe pump to perform the extraction. The microsyringe was used as both the microextraction device as well as the sample introduction device for GC-MS analysis. The attached hollow fiber served as the "holder" and protector" of 3 microl of organic solvent. The solvent was repeatedly withdrawn into and discharged from the hollow fiber by the syringe pump. Pesticides were extracted from 4-ml water samples into the organic solvent impregnated in the hollow fiber. The effects of organic solvents, plunger movement pattern, agitation and extraction time were investigated. Good repeatabilities of extraction performance were obtained, with the RSD values ranging from 3.0% (alachlor) to 9.8% (4-chlorophenol) for the 14 pesticides; most RSD values were under 5.0%. The method provided a 490-fold preconcentration of the target pesticides. The limits of detection were in the range of 0.01-5.1 microg/l (S/N = 3) in the GC-MS selected ion monitoring mode. In addition, sample clean-up was achieved during LPME because of the selectivity of the hollow fiber, which prevented undesirable large molecules from being extracted. A slurry sample (mixture of 40 mg soil/ml of water) containing seven pesticides was extracted using this method which also gave good linearity and precision (most RSDs <7.0%, n = 3). PMID- 12580478 TI - Comparison of solid-phase microextraction and purge-and-trap methods for the analysis of the volatile fraction of butter. AB - The volatile fraction of butter stored at three different temperatures was investigated to monitor quality during commercial shelf-life (90 days). Two different extraction techniques were compared: dynamic headspace (purge-and trap), and static headspace (solid-phase microextraction, SPME). As expected, the dynamic extraction provided a generally higher amount of volatile compounds than that obtained by SPME, but, with reference to individual compounds, SPME seemed to provide better extraction for volatiles having a higher molecular mass. Despite the different performances, both methods were able to detect volatiles useful for evaluating changes during storage. PMID- 12580479 TI - Monitoring and fast detection of mycotoxin-producing fungi based on headspace solid-phase microextraction and headspace sorptive extraction of the volatile metabolites. AB - Solid phase microextraction in combination with capillary GC-MS was used as monitoring technique for the collection and detection of the fungal volatile metabolite (+)-aristolochene by sporulated surface cultures of Penicillium roqueforti. A comparison was made between different toxigenic and nontoxigenic strains of P. roqueforti. Different growth conditions and media, such as malt extract agar, potato dextrose agar and sabouraud dextrose agar were compared. Whereas toxigenic strains produced large amounts of (+)-aristolochene, beta elemene, valencene and germacrene A, nontoxigenic P. roqueforti strains showed a remarkably different headspace profile, in which ethyl-2-hexenoate, E-beta caryophyllene, aromadendrene and beta-patchoulene were the predominant volatiles, apart from other sesquiterpene hydrocarbons present at lower concentrations. Stir bar sorptive extraction, was also applied in the headspace sampling mode, i.e. headspace sorptive extraction (HSSE) for the enrichment of fungal volatiles from sporulated surface cultures to differentiate between toxigenic and nontoxigenic fungi. Hence, it can be concluded that headspace analysis of volatile fungal metabolites by SPME and HSSE in combination with capillary GC-MS is a suitable monitoring technique for the fast detection of mycotoxin producing fungi. PMID- 12580481 TI - Development of a gas chromatography-mass spectrometry method for the determination of household insecticides in indoor air. AB - This work presents a GC-MS method for the determination of 17 household insecticides and acaricides in indoor air. Air samples were collected with a sampling train which consisted of a glass fibre filter and two polyurethane foam plugs, followed by a high-volume air pump. Filters and plugs were analysed separately. The overall recoveries ranged from 85 to 109% (4-11% RSD). Minimum method detection limits between 0.1 and 5 ng/m3 were determined. PMID- 12580480 TI - Optimization of a microwave-assisted extraction method for the analysis of polychlorinated biphenyls in ash samples. AB - An alternative method for the extraction of polychlorinated biphenyls (PCBs) in ash samples, which is less time and solvent consuming than Soxhlet extraction, is presented. A study was carried out to evaluate the possibilities of microwave assisted extraction (MAE) to determine exactly which parameters affect the efficiency of the process, since direct extrapolation of extraction conditions for PCBs in other solid matrices, failed when applied to coplanar congeners in ash samples. Influence of the organic solvent on the yield of the extraction was first evaluated using two ash samples with different percentages of carbon. Once the extraction solvent was fixed, the effects of solvent volume, extraction temperature and extraction time were investigated using an experimental design. It was found that the volume of organic solvent played a more important role in the extraction efficiency than the other factors. In the optimal conditions microwave extractions were performed at 110 degrees C. for 10 min and using 30 ml of toluene. Recoveries higher than 80% were obtained for all the highly chlorinated congeners. including coplanar species, in a spiked ash sample containing a relatively high concentration of carbon. The proposed method was also applied to the determination of PCBs in a reference material of urban dust. Recoveries were similar to those obtained for spiked ash samples. PMID- 12580482 TI - Fast supercritical fluid extraction and high-resolution gas chromatography with electron-capture and flame photometric detection for multiresidue screening of organochlorine and organophosphorus pesticides in Brazil's medicinal plants. AB - A description is given of a rapid and environmentally friendly method to determine organochlorine and organophosphorus pesticide multiresidues--malathion, methidathion, fenitrothion, fenthion, parathion-ethyl, parathion-methyl, lindane, hexachlorobenzene, chlorothalonil, tetradifon, alpha-endosulfan, beta-endosulfan and dieldrin-in Passiflora alata Dryander and pasiflora edulis Sims. f. flavicarpa Deg. leaves by supercritical fluid extraction and high-resolution gas chromatography with electron-capture and flame photometric detection (HRGC ECD/FPD). The mild extraction conditions [pure CO2; 100 bar (1 bar = 10(5) Pa) and 40 degrees C (p = 0.62 g/ml); 5 min static+10 min dynamic extraction time; ODS trap and elution with 1 ml n-hexane at 2 ml/min] allow for direct analysis by HRGC-ECD/FPD with no prior cleaning procedure. The method provides. in accordance with the validation criteria of the European Pharmacopoeia, analytical results that are similar or even better than the official procedures, and is simpler, faster and cheaper. Mean recoveries of 69.8-107.1% were obtained, with 1.4-14.7% reproducibility (RSD). The method was applied to analyse commercial samples of Passiflora L. from Brazil. Twenty-three percent of the samples showed the presence of the organochlorine or organophosphorus pesticide residue investigated. PMID- 12580483 TI - Determination of triazines in soil by microwave-assisted extraction followed by solid-phase microextraction and gas chromatography-mass spectrometry. AB - A method for determining triazine herbicides in soil samples that combines microwave-assisted extraction with solid-phase microextraction is described. Water containing 1% methanol was employed as extractant. The parameters of solid phase microextraction and microwave-assisted extraction were investigated. In solid-phase microextraction, particular attention was paid to the negative effect of salt on fiber stability. Our experiments showed that this effect could be effectively reduced by simply washing the fiber with deionized water. The selected triazines could be efficiently extracted by the aqueous extractant at 105 degrees C for 3 min, with 80% output of maximum power (1,200 W). The extraction procedure provided good precision (<7%) and recoveries (76.1-87.2%). The limits of detection were in the range 2-4 microg/kg. Compared with conventional liquid extraction, microwave-assisted extraction-solid-phase microextraction was more efficient, accurate and faster, and used a very small amount of organic solvent (only 250 microL methanol). The extraction of aged spiked soil samples indicated that, although the recoveries were lower than those of freshly spiked samples, they were nevertheless satisfactory for the quantitative analysis of real-world samples. PMID- 12580484 TI - On-fiber photodegradation after solid-phase microextraction of p,p'-DDT and two of its major photoproducts, p,p'-DDE and p,p'-DDD. AB - The potential of performing photochemical studies in solid phase microextraction (SPME) fibers, "photo-SPME", to study the photodegradation of p,p'-DDT and two of its major degradation products, p,p'-DDE and p,p'-DDD, is shown. Analyses were carried out by gas chromatography mass spectroscopy detection. DDT was extracted from aqueous solutions using five different commercial coatings. The fibers were then exposed to UV light emitted by a low-pressure mercury lamp. After 30 min of irradiation, the degradation of DDT only occurred in polydimethylsiloxane fibers. The on-fiber degradation kinetics of p,p'-DDT was studied from 2 to 60 min. A large number of photoproducts were generated and their kinetic behavior was studied. In order to clarify the possible photoreaction pathways for DDT, individual water solutions containing p,p'-DDD or p.p'-DDE were prepared and photo-SPME was performed for each compound at different irradiation times. On the basis of the photoproducts identified, some photodegradation pathways are proposed. Finally, aqueous photodegradation studies followed by SPME were performed and compared to the photo-SPME. This work will show the enormous potential of photo-SPME to perform photodegradation studies. PMID- 12580485 TI - Lichens as polycyclic aromatic hydrocarbon bioaccumulators used in atmospheric pollution studies. AB - The aim of this work has been to determine the possibility of using lichens as polycyclic aromatic hydrocarbon (PAH) bioaccumulators for the evaluation of atmospheric pollution in the city of Rieti. A lichen sample collected in a remote unpolluted area was divided into 18 sub-samples. These were collocated in nine different stations in the city of Rieti and every 5 months the PAH concentrations were determined. Only phenanthrene, anthracene, fluoranthene, pyrene and chrysene were found at concentrations over the limit of quantification. In the meantime a continuous increase of their concentrations was observed. PMID- 12580486 TI - Trace analysis of benzene, toluene, ethylbenzene and xylene isomers in environmental samples by low-pressure gas chromatography-ion trap mass spectrometry. AB - A rapid determination of benzene, toluene, ethylbenzene and the three xylene isomers (BTEX), including a nearly baseline separation of the xylene isomers in environmental samples within 1 min has been carried out using low-pressure gas chromatography-ion trap mass spectrometry (LP-GC-IT-MS). In order to evaluate the different parameters which may influence the performance of LP-GC-IT-MS, different column and mass spectral parameters were varied. Comparing LP-GC-IT-MS with the conventional equivalent, we obtained excellent detection limits as well as a good RSD of 8-13% in ition to a much shorter analysis time. In order to evaluate LP-GC-IT-MS for use in environmental samples, we determined BTEX in air. PMID- 12580487 TI - Determination of total and polycyclic aromatic hydrocarbons in aviation jet fuel. AB - The aviation jet fuel widely used in turbine engine aircraft is manufactured from straight-run kerosene. The combustion quality of jet fuel is largely related to the hydrocarbon composition of the fuel itself; paraffins have better burning properties than aromatic compounds, especially naphthalenes and light polycyclic aromatic hydrocarbons (PAHs), which are characterised as soot and smoke producers. For this reason the burning quality of fuel is generally measured as smoke fermation. This evaluation is carried out with UV spectrophotometric determination of total naphthalene hydrocarbons and a chromatographic analysis to determine the total aromatic compounds. These methods can be considered insufficient to evaluate the human health impact of these compounds due to their inability to measure trace (ppm) amounts of each aromatic hyrcarbon and each PAH in accordance with limitations imposed because of their toxicological properties. In this paper two analytical methods are presented. Both are based on a gas chromatographic technique with a mass detector operating in be selected ion monitoring mode. The first method was able to determine more than 60 aromatic hydrocarbons in a fuel sample in a 35-min chromatographic run, while the second was able to carry out the analysis of more than 30 PAHs in a 40-min chromatographic run. The linearity and sensitivity of the methods in measuring these analytes at trace levels are described. PMID- 12580488 TI - Determination of butyltin compounds in surface sediments from the Sao Paulo State coast (Brazil) by gas chromatography-pulsed flame photometric detection. AB - Occurrence and the effects of butyltin compounds (BTs) have been studied for some years, mainly in countries of the Northern Hemisphere. Due to widespread use of tributyltin compounds (TBTs) and considering their deleterious effects, it is necessary to conduct studies on its occurrence, especially in the marine environment because of its excessive use in coatings of ship hulls to prevent fouling. Moreover, it was important to extend the evaluation to areas where there is no current information about their occurrence. The present work reports the occurrence of BTs in marine sediments of Sao Paulo state, Brazil. Commercial and leisure harbor sampling sites were selected because these areas are potentially exposed to BTs from antifouling paints used on ship hulls. Analytical conditions for organotin analysis in marine sediments were optimized for GC with pulsed flame photometric detection. Detection limits ranged from 8A to 66.3 ng g(-1) using a 610-nm filter, and the linearity range was 20-500 ng g(-1). Concentration levels of BTs were highest in Santos harbor (360 ng g(-1) TBT in average) and Guaruja marina (670 ng g(-1) TBT in average), which seems to be related to intensive boat traffic. Lower levels of BTs were observed in Cananeia, where only fishing boats are present (50 ng g(-1) TBT in average). PMID- 12580489 TI - Solid-phase extraction-thin-layer chromatography-gas chromatography method for the detection of hazelnut oil in olive oils by determination of esterified sterols. AB - The sterol composition of extra virgin olive oil is very characteristic and, thus, has become a helpful tool to detect adulterations with other vegetable oils. Special attention has been addressed to the separate determination of the free and esterified sterol fractions, since both have different compositions and can thus provide more precise information about the actual origin of the olive oil. In the case of admixtures with small amounts of hazelnut oil, this approach can be extremely useful, because the similarity between the fatty acid compositions of both oils hampers the detection of the fraud. A hyphenated chromatographic method was developed for a sensitive and precise determination of esterified sterols in olive oils. The oil was subjected to silica solid-phase extraction (SPE) fractionation, cold saponification of the collected fraction and purification on silica TLC. The sterol band was then injected into an SPB-5 (30 m x 0.25 mm I.D., 0.25 microM film thickness) and the ratio [% campesterol x (% 7 stigmastenol)2]/(% 7-avenasterol) was calculated. The method was tested on extra virgin olive oil; good sterol recoveries and repeatability were obtained. The results were compared with another method. which has a different sample preparation sequence (silica column chromatography, hot saponification and silica TLC). Similar results were achieved with both methods; however, the SPE-cold saponification-TLC-capillary GC was faster, required less solvent and prevented sterol decomposition. The SPE-method was applied to an admixture with 10% of hazelnut oil and to a screening of 11 oils (husk oil, virgin and refined olive oils) from different Mediterranean countries. PMID- 12580490 TI - Analysis of volatiles of malt whisky by solid-phase microextraction and stir bar sorptive extraction. AB - Blended Scotch whisky was analysed by solid-phase microextraction (SPME) and stir bar sorptive extraction (SBSE) to study the composition of the volatiles. For SPME analysis, three different fibres were compared, poly(dimethylsiloxane) (PDMS) (100 microm). poly(acrylate) (PA) (85 microm) and divinylbenzene-Carboxen on poly(dimethylsiloxane) (DVB-CAR-(PDMS) (50/30 microm). It was found that the PDMS and DVB-CAR-PDMS fibres showed a higher enrichment capacity than PA as well as a better reproducibility. The influence of sampling time, temperature and salt addition on the enrichment of volatiles as well as the difference between liquid and headspace SPME were studied. An optimum SPME method was developed. Finally a more recent sample preparation technique, namely SBSE was evaluated to extract whisky volatiles. PMID- 12580491 TI - Flavour analysis of Greek white wine by solid-phase microextraction-capillary gas chromatography-mass spectrometry. AB - Solid-phase microextraction (SPME) was optimised for the qualitative determination of the volatile flavour compounds responsible for the aroma of Greek Boutari wine. Several factors influencing the equilibrium of the aroma compounds between the sample and the SPME fiber were taken into account, including the extraction time, the extraction temperature, the sampling mode (headspace and direct immersion or liquid SPME), and the presence of salt. Four different SPME fibers were used in this study. namely poly(dimethylsiloxane) (PDMS), poly(acrylate), carbowax-divinylbenzene and divinylbenzene-carboxen on poly(dimethylsiloxane). The best results were obtained using the PDMS fiber during headspace extraction at 25 degrees C for 30 min after saturating the samples with salt. The optimised SPME method was then applied to investigate the qualitative aroma composition of three other Greek wines, namely Zitsa, Limnos and Filoni. PMID- 12580492 TI - Direct quantitation of volatile organic compounds in packaging materials by headspace solid-phase microextraction-gas chromatography-mass spectrometry. AB - The quantification of volatile organic compounds (VOCs) in flexible multilayer packaging materials using headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) was studied. The analytes imclude 22 compounds such as aldehydes. ketones, carboxylic acids and hydrocarbons formed by thermooxidative degradation of polyethylene during the extrusion coating process in the manufacture of the packaging, and many of them are involved in the unpleasant and undesirable odour of these materials. External standard calibration using a solution of the analytes in an appropriate solvent was the first approach studied. Aqueous solutions of the analytes provided low reproducibility and the reduction of aldehydes to alcohols under the HS-SPME conditions. Hexadecane was chosen as the solvent since its polarity is similar to that of polyethylene and its volatility is lower than that of the analytes. However, hexadecane should be added to the sample before the analysis as it modifies the absorption capacity of the fibre. A 75-microm Carboxen poly(dimethylsiloxane) fibre was used to extract the VOCs from the headspace above the packaging in a 15-ml sealed vial at 100 degrees C after 5 min of preincubation. The influence of the extraction time on the amount extracted was studied for a standard solution of the analytes in hexadecane, together with the influence of the volume of the standard solution and the amount of the sample placed in the vial. Standard addition and multiple HS-SPME were also studied as calibration methods and the results obtained in the quantitative analysis of a packaging material were compared. PMID- 12580493 TI - Enflurane as an internal standard in monitoring halogenated volatile anaesthetics by headspace gas chromatography-mass spectrometry. AB - Recently. we proposed the use of a run-only headspace-GC-MS method for the biological monitoring of ppb concentrations of unmodified volatile anaesthetics (isoflurane, sevoflurane and halothane, plus nitrous oxide) in post-shift urine of operating theatre personnel. The adoption of enflurane (a volatile anaesthetic no longer used in clinical practice) as a poper and viable internal standard improves intra-day and inter-day accuracy in halide quantitation, providing a GC MS reference method useful in the practice of biomonitoring of exposure of operating theatre personnel to modern volatile anaesthetics (isoflurane. sevoflurane, halothane). PMID- 12580494 TI - Determination of acidic drugs in sewage water by gas chromatography-mass spectrometry as tert.-butyldimethylsilyl derivatives. AB - A procedure is described for the determination of five acidic non-steroidal anti inflammatory pharmaceuticals (ibuprofen, naproxen, ketoprofen, tolfenamic acid and diclofenac) in sewage water. The analytical method involves the concentration of water samples using a solid-phase extraction polymeric sorbent, functionalized with N-vinylpyrrolidone. Analytes were eluted with ethyl acetate. derivatized using N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide (MTBSTFA) and analyzed by GC-MS. Influence of time, temperature and volume of MTBSTFA in the yield of the derivatization step were studied in detail using a factorial central composite design. Quantification limits of the analytical procedure for 500 ml of sewage water ranged from 20 to 50 ng/l. Recoveries from 90 to 115% were found for sewage water samples spiked with the studied compounds at the low ng/ml level. Results obtained for real samples show the presence of ibuprofen and naproxen in both influent and effluent of a sewage water treatment plant. PMID- 12580495 TI - Gas chromatography with mass spectrometric, atomic emission and Fourier transform infrared spectroscopic detection as complementary analytical techniques for the identification of unknown impurities in pharmaceutical analysis. AB - An example of the complementary use of GC-MS. GC-AED and GC-FT-IR is described for efficient structure elucidation of an unknown impurity in pharmaceutical analysis. None of the analytical techniques could solve the structure of the unknown impurity alone; identification was, however, straightforward by combining the available spectroscopic information. GC-MS provided information about structural fragments and molecular mass of the unknown compound. GC-AED was used for confirmation of the occurrence of the individual elements in the structure and to enable calculation of the empirical formula. GC-FT-IR gave valuable information regarding functional groups in the molecule. PMID- 12580496 TI - Relaxed eddy accumulation, a new technique for measuring emission and deposition fluxes of volatile organic compounds by capillary gas chromatography and mass spectrometry. AB - The possibility afforded by a new relaxed eddy accumulation system in the determination of emission and deposition fluxes of volatile organic compounds (VOCs) by chromatographic techniques will be presented. The system, especially designed to limit sampling artifacts, uses adsorption traps filled with solid sorbents as reservoirs for VOC collection. Enriched compounds were analyzed by capillary GC and positive identification and quantification of eluted compounds was Achieved by mass-spectrometric detection. The method has been used to quantify the emission and deposition of both biogenic and anthropogenic VOCs over a Mediterranean forest ecosystem located in Central Italy. For the first time, both daily and seasonal trends of anthropogenic and biogenic VOCs will be reported. The consistency of monoterpene fluxes with predictions based on the knowledge of VOC emission from the dominant vegetation species will be tested. Data have been used to develop a novel algorithm to predict the seasonality of biogenic emission from the forest ecosystem. PMID- 12580497 TI - Headspace gas chromatography-mass spectrometry of volatile compounds in murici (Byrsonima crassifolia l. Rich). AB - Northern and Northeastern Brazil have a natural diversity of fruits, many of which are considered exotic, presenting different flavors and aromas. The enormous diversity of fruits represents a promising area for research on aromas. There is also a great potential for the manufacture of juices, desserts or other processed products. Murici is a typical fruit from these regions presenting a different flavor, reminiscent of that of cheese. This fruit is consumed mainly as juice, ice cream or as liquor, greatly appreciated by the local population. Headspace volatile compounds of three lots of the fruit from Ceara (Fortaleza) were collected by suction on Porapak Q for 2 h and desorbed with 300 microl of acetone. The isolated volatile compounds were separated by high resolution GC. Forty-six volatile compounds were detected, of which 41 were identified by gas chromatography-mass spectrometry and Kovats indices. The most abundant compounds were ethanol (28.3%) and ethyl hexanoate (25.1%). Butanoic acid (5.1%), hexanoic acid (5.1%) and methyl butyrate (2.8%) were also detected in the headspace of the fruit and confirm its unusual cheese aroma. PMID- 12580498 TI - Analysis of pyridines in mainstream cigarette smoke. AB - A new technique has been developed for the quantitative analysis of pyridines in mainstream cigarette smoke using a GC-MS technique. For analysis, 10 cigarettes are smoked using conditions based on US Federal Trade Commission recommendations. The smoke is collected in a water trap and analyzed using a GC-MS technique. A standard or a fast GC separation can be applied for the analysis. The standard separation was followed by MS detection using selected ion monitoring (SIM) acquisition on a quadrupole instrument. The fast GC was followed by MS detection with total ion acquisition on a time-of-flight instrument. The levels of pyridine depend on the type of cigarette: for a full flavor cigarette pyridine is as high as 18.0 microg/cigarette (cig.). and for an ultra light cigarette is about 3.0 microg/cig. Substituted pyridines vary between 5.0 microg/cig. to 0.1 microg/cig. for a full flavor cigarette, and between 0.2 microg/cig. and a few ng/cig. for an ultra light cigarette. The reproducibility of the technique is very good, with less than 7-8% RSD in both separation procedures for most of the analyzed compounds. PMID- 12580499 TI - Determination of the disproportionation products of limonene used for the catalytic hydrogenation of castor oil. AB - The direct determination by GC-MS analysis of the products from the limonene disproportionation obtained from the catalytic hydrogenation of castor oil is not possible since a transesterification of the oil is required before injection. Therefore, a method for the determination of those products is proposed here: a solid-phase microextraction (SPME) followed by GC-MS. With the optimization of SPME it was possible to isolate a great number of products of the disproportionation of limonene used as hydrogen donor in the catalytic hydrogenation of castor oil by Pd/C. This system also proved to be appropriate to monitor castor oil hydrogenation by following and identifying the limonene disproportionated products. PMID- 12580500 TI - Chiral separation of gamma-butyrolactone derivatives by gas chromatography on 2,3 di-O-methyl-6-O-tert.-butyldimethylsilyl-beta-cyclodextrin. AB - The chiral GC separation of 2-alkyl-2-keto-gamma-butyrolactone derivatives and their alcohol analogs using 2,3-di-O-methyl-6-O-tert.-butyldimethylsilyl-beta cyclodextrin (DIMETBCD) as chiral selector was discussed. The results, supported by the ketone preliminary molecular modelling calculations, suggest that the chiral recognition for DIMETBCD depends more on the geometry than on the polarity of the alkyl substituents on the butyrolactones. Hydrogen bonds and alkyl group steric effects should be an important function of the alcohol chiral recognition for DIMETBCD. Comparison of the retention times of the alcohol derivatives, in achiral and chiral stationary phases, suggests a specific structural effect for the cyclodextrin selctor. PMID- 12580501 TI - Analysis of the oxidation products of cis- and trans-octadecenoate methyl esters by capillary gas chromatography-ion-trap mass spectrometry. I. Epoxide and dimeric compounds. AB - The oxidative behavior of methyl oleate (MeOl) and methyl elaidate (MeEl) was compared by hyphenated chromatographic techniques. MeOl and MeEl were separately oxidized (200 degrees C/30 min) and subjected to solid-phase extraction, in order to isolate the low polarity compounds. The two isomeric 9,10-epoxystearic methyl esters formed in both MeOl and MeEl at different threolerythro ratios (2.3 and 6.2%, respectively). The dimeric products produced in the thermoxidized MeOl and MeEl (1.4 and 1.6%, respectively), showed similar gas chromatographic characteristics and mass spectra, suggesting similar molecular structures and formation mechanisms. A positional and probably configurational mixture of symmetric and asymmetric dehydrodimers was detected, whereas the occurrence of MeEl or MeOl dimeric ethers is to be confirmed. PMID- 12580502 TI - Quantitative structure-retention relationships of phenolic compounds without Hammett's equations. AB - Retention times of phenolic compounds in a given pH eluent in reversed-phase liquid chromatography were predicted from dissociation constants derived from atomic partial charges and log P-values calculated by a computational chemical method. The precision of the calculation of atomic partial charges by AMI and PM3 methods of MOPAC was evaluated. The atomic partial charges obtained by AMI were the more acceptable. The atomic partial charges obtained from the hydrogen of the hydroxyl group included an ortho-effect, therefore an ortho-effect was added to the predicted values. The precision of predicted retention factors obtained using predicted pKa values was similar to that using reference pKa values. PMID- 12580503 TI - Monolithic silica column for in-tube solid-phase microextraction coupled to high performance liquid chromatography. AB - In-tube solid-phase microextraction (SPME) has successfully been coupled to capillary LC, and further an automated in-tube SPME system has been developed using a commercially available HPLC auto-sampler. However, an open tubular capillary column with a thick film of polymer (stationary phase) is unfavorable because the ratio of the surface area of coating layer contacted with sample solution to the volume of the capillary column is insufficient for mass transfer. A highly efficient SPME column is. therefore, required. We introduced a C18 bonded monolithic capillary column that was used for in-tube SPME. The column consisted of continuous porous silica having a double-pore structure. Both the through-pore and the meso-pore were optimized for in-tube SPME, and the optimized capillary column was connected to an HPLC injection valve for characterization. The results demonstrated that the pre-concentration efficiency is excellent compared with the conventional in-tube SPME. The novel method for both introduction and concentration of the samples was effective. satisfactory and suitable for use in the SPME medium. PMID- 12580504 TI - Ion chromatography on reversed-phase materials coated with mixed cationic and nonionic surfactants. AB - Columns suitable for use in anion chromatography can be prepared by coating a packed reversed-phase HPLC column (C18 silica or polystyrene particles) with a cationic surfactant. The efficiency is improved dramatically by first coating the column with a nonionic surfactant and then subsequently with the cationic surfactant. The thickness of the first coated layer as well as the chemical structure of the surfactant have a major effect on the column performance. Actual separations are included to demonstrate the convenience and practical use of the coated columns. Using this approach, columns with 12,900 theoretical plates for the 15-cm column (or 86,000 plates/m) were produced, giving well shaped peaks with an average asymmetry factor of 1.09. The coated layers were found to be stable, giving retention times with an average relative standard deviation of 1.6% for 12 consecutive runs. PMID- 12580505 TI - Preliminary characterization of anhydrous ethanol used in Brazil as automotive fuel. AB - This work reports preliminary studies on the characterisation of anhydrous ethanol (AEA) used as an automotive fuel mixed with gasoline in Porto Alegre (South Brazil). Pre-concentration of the impurities contained in 1000 ml of AEA was carried on through solid-phase extraction using XAD4 resin. The main compounds in the extract were identified by means of spectral data from the library of the equipment. The concentrate was then fractionated using a preparative liquid chromatographic column filled with activated silica gel and the elution procedure was carried out with, n-hexane, n-hexane-benzene (1:1, v/v) and dichloromethane. Prior to analysis by GC-MS, each fraction was reduced to 1 ml with a gentle stream of nitrogen. Saturated linear hydrocarbons and aromatic hydrocarbons eluted in the first fraction and oxygenated compounds such as aldehydes. ketones and alcohols, eluted in the second one. were the main compounds detected in the sample. PMID- 12580506 TI - Improvement in steroid screening for doping control with special emphasis on stanozolol. AB - The Medical Commission of the International Olympic Committee forbids the use of anabolic androgenic steroids and beta2-agonists to improve athletic performance. In this work we have selected examples of anabolic androgenic compounds and their metabolites to evaluate the GC-MS analysis of some trimethylsilyl derivatives. The aim is to set the best GC conditions to improve the detection within the whole range of analyte elution temperatures. The initial column temperature was changed to 105 or 140 degrees C followed by 40 degrees C min(-1) to 200 degrees C and then 15 degrees C min(-1) to 300 degrees C. Using 140 degrees C as the initial oven temperature it was possible to obtain narrower initial analyte distributions for the compounds that elutes at the beginning of the chromatogram as clenbuterol, mabuterol, epimethylenediol and norandrosterone, without loss of derivatized metabolites signal. Later. eluting analytes, such as the stanozolol metabolites, furazabol and oxandrolone were not affected. Temperatures below 140 degrees C. resulted in partial derivatization for some analytes mainly stanozolol related structures. Therefore evaluation of derivatization conditions as occurring in three steps, the vial, vaporization chamber and capillary column, was thoroughly assessed. The new program temperature improves the signal-to-noise ratio for some compounds and shows adequate resolution for endogenous compounds. Some of the difficult key separations necessary for doping control enforcement were also obtained with the proposed method. PMID- 12580507 TI - Ginkgo biloba extract preserves pyruvate and enhances ascorbate in the cortex of gerbils during focal cerebral ischemia. A microdialysis-liquid chromatography study. AB - The aim of this study was to evaluate dynamic changes in energy-related metabolites in the cortex of gerbils subjected to focal cerebral ischemia after pretreatment with Ginkgo biloba extract. Focal cerebral ischemia was induced by occlusion of the right common carotid artery and the right middle cerebral artery for 60 min in anesthetized gerbils. A microdialysis probe was inserted into the cortex to monitor extracellular lactate. pyruvate and ascorbate during ischemia and reperfusion. The present study demonstrated a dynamic decrease in pyruvate (25% of baseline) and increases in lactate (160% of baseline) and asorbate (300% of baseline) and a 5-fold increase in the lactate:pyruvate (L:P) ratio during cerebral ischemia in the control group. However. pyruvate levels were preserved and ascorbate levels were enhanced with a chronic pretreatment of Ginkgo biloba extract for 8 days (i.p., 100 mg kg(-1) day(-1)). Preservation of pyruvate and enhancement of ascorbate observed in this study may be associated with the neuroprotective effects of Ginkgo biloba extract. PMID- 12580508 TI - Determination of E-2-nonenal by high-performance liquid chromatography with UV detection assay for the evaluation of beer ageing. AB - The analysis of E-2-nonenal is of considerable interest for the brewery industry as this compound is claimed to be responsible for a paper/cardboard unpleasant flavour. Usually, the presence of E-2-nonenal can be noticed in aged beers at levels higher than 0.1 microg/l. In this work, an analytical method was developed to determine E-2-nonenal in beer involving steam distillation of beer followed by an extraction/concentration step using solid-phase extraction and determination of E-2-nonenal by HPLC with UV detection. Fastness and simplicity are the main advantages of the proposed method, when compared with other existing methodologies for the determination of E-2-nonenal in beer. Using the developed conditions, the interference of E-2-nonenal formed by degradation of its precursors during steam distillation is almost negligible. The presence of sulphur dioxide at legal levels does not interfere with the assay. The method was used in a comparative study of fresh and either naturally or forced aged beers. A much larger chromatographic peak was found near the peak of E-2-nonenal that correlates well with the peak of E-2-nonenal. Identification of the corresponding compound is currently under investigation, considering its future application on the evaluation of beer ageing. PMID- 12580509 TI - Coupled liquid chromatography-gas chromatography for the rapid analysis of gamma oryzanol in rice lipids. AB - An approach based on on-line coupled liquid chromatography-gas chromatography (LC GC) was developed for the rapid analysis of gamma-oryzanol in rice. Total lipids were extracted from rice and subjected to LC-GC without any prior purification. gamma-Oryzanol was pre-separated by HPLC from rice lipids and transferred on-line to GC analysis in order to separate its major constituents. 24 methylenecycloartanyl ferulate, cycloartenyl ferulate, campesteryl ferulate, beta sitosteryl ferulate and campestanyl ferulate. The identities of the compounds were confirmed by off-line GC-MS analysis. Total gamma-oryzanol content could be quantified by HPLC-UV detection and the distribution of gamma-oryzanol constituents could be determined by on-line coupled GC analysis. The proposed methodology paves the way for high-throughput investigations providing information on natural variations in gamma-oryzanol content and its composition in different rice varieties. PMID- 12580510 TI - Evaluation of chromatographic versus electrophoretic behaviour of a series of therapeutical peptide hormones. AB - In this work, models describing the effect of pH on chromatographic and electrophoretic behaviour for a series of polyprotic therapeutic peptide hormones were compared. taking into account the species in solution and the activity coefficients. The usefulness of the proposed equations is twofold, they permit the determination of the acidity constants in water and in the hydroorganic mobile phases used in liquid chromatography (LC) and capillary electrophoresis (CE) and can also be used for the selection of the optimum pH for the separation of mixtures of the modelled compounds. The proposed relationships allow an important reduction of the experimental data needed for the development of new separation methods. The accuracy of the proposed equations is verified by modelling the chromatographic and electrophoretic behaviour of a series of polyprotic therapeutic peptide hormones. By calculating the values of predicted resolutions, selection of the optimum pH to perform LC or CE separations of their mixtures becomes a rapid and simple process. PMID- 12580511 TI - Liquid-liquid and solid-phase extractions of phenols from virgin olive oil and their separation by chromatographic and electrophoretic methods. AB - The high oxidative stability of virgin olive oil is related to its high monounsaturated/polyunsaturated ratio and to the presence of antioxidant compounds, such as tocopherols and phenols. In this paper, the isolation of phenolic compounds from virgin olive oil, by different methods, was tested and discussed. Particularly liquid-liquid and solid-phase extraction methods were compared, assaying, for the latter, three stationary phases (C8, C18 and Diol) and several elution mixtures. Quantification of phenolic and o-diphenolic substances in the extracts was performed by the traditional Folin-Ciocalteau method and the sodium molybdate reaction, respectively. Furthermore, the quantification of phenolic compounds in the extracts and in a standard mixture was carried out both with diode array and mass spectrometric detection and capillary zone electrophoresis. PMID- 12580512 TI - On-line sample preconcentration in micellar electrokinetic chromatography by sweeping with anionic-zwitterionic mixed micelles. AB - On-line preconcentration by sweeping in micellar electrokinetic chromatography using mixed micelles of sodium dodecyl sulfate (SDS)-SB-12 is presented. Because of their large micelle radius, they permit increased partitioning of hydrophobic analytes into the core. In addition, they also possess lower negative surface charge relative to pure SDS micelles so anionic analytes can be retained better due to decreased electrostatic repulsion. As the efficiency of sweeping is predicated on the magnitude of retention factors, these advantages translated to better focusing. As much as a 370-fold improvement in detector response, in terms of peak height, was obtained for some neutral steroids, while about a 360-fold improvement was obtained for some phenol derivatives, which were previously not amenable to sweeping by pure SDS micelles. PMID- 12580513 TI - 1,3-dialkylimidazolium-based room-temperature ionic liquids as background electrolyte and coating material in aqueous capillary electrophoresis. AB - The 1-ethyl-3-methylimidazolium (EMIM) cation was found to have constant mobility of 4.5 x 10(-4) cm2 V(-1) s(-1) over the pH range of 3 to 11. The electroosmotic flow of bare silica capillary was reversed by the covalently bonded room temperature ionic liquid (RTIL) coating. With run buffer of 5 mM EMIM (pH 8.5), NH4+ in human urine was separated from the K+ matrix and was detected to be 0.37 +/- 0.012%. K+, Na+, Li+, Ca2+, Mg2+ and Ba2+ were baseline separated in RTIL coated capillary with run buffer of 10 mM EMIMOH-acetic acid at pH 5, and the concentration of the above ions in a red wine were detected to be 907, 27.9, 0, 71.0, 83.4 and 31.1 microg/ml, respectively. The RTIL-coated capillary showed stable electroosmotic flow for at least 80 h in the run buffer. PMID- 12580514 TI - Voltage-controlled separation of proteins by electromobility focusing in a dialysis hollow fiber. AB - Electromobility focusing (EMF) is a relatively new protein separation technique that utilizes an electric field gradient and a hydrodynamic flow. Proteins are focused in order of electrophoretic mobility at points where their electrophoretic migration velocities balance the hydrodynamic flow velocity. Steady state bands are formed along the separation channel when equilibrium is reached. Further separation and detection can be easily achieved by changing the electric field profile. In this paper. we describe an EMF system with on-line UV absorption detection in which the electric field gradient was formed using a dialysis hollow fiber. Protein focusing and preconcentration were performed with this system. Voltage-controlled separation was demonstrated using bovine serum albumin and myoglobin as model proteins. The limitations of the current method are discussed, and possible solutions are proposed. PMID- 12580515 TI - Determination of free L- and D-alanine in hydrolysed protein fertilisers by capillary electrophoresis. AB - of racemisation of hydrolysed protein fertilisers (HPFs) using an The objective of this study was to determine the degree inexpensive and easy to handle analytical method for qualitative control of the products. Using a polyacrylamide coated capillary and a run buffer containing 0.1 M Tris-borate+2.5 mM EDTA Na2+0.1% sodium dodecylsulfate+10 mM beta-cyclodextrin a quantitative separation of D- and L-alanine (Ala) was made from an not treated HPF sample derivatised with dansyl chlorine by capillary electrophoresis. The D-Ala:[D-Ala+L-Ala] ratio, called degree of racemisation (RD), was calculated. The analysis of ten commercial HPFs has shown that more than 60% of HPFs have an RD > or = 40%. while only one product has shown an RD <5%. These results showed that most of the HPFs on the market are obtained with strong hydrolytic processes and high contents of D-amino acids are probably less effective as plant nutrients or even potentially dangerous to plants. PMID- 12580516 TI - Separation of conjugated trienoic fatty acid isomers by capillary electrophoresis. AB - A method for direct resolution of conjugated trienoic fatty acid isomers by capillary electrophoresis has been developed. To obtain complete separation a dual cyclodextrin system was used. This contained heptakis-(6-sulfo)-beta cyclodextrin (charged). Beta-cyclodextrin (uncharged) and sodium dodecylsulfate. Under optimized conditions, all seven isomers were well separated. On average, separation efficiency was 2.9 x 10(5) plates/m. PMID- 12580517 TI - Characterization of polyethylene glycols and polypropylene glycols by capillary zone electrophoresis and micellar electrokinetic chromatography. AB - Methods based on capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) have been developed and optimized for the separation of polyethylene glycols (PEGs) and polypropylene glycols (PPGs). To provide for charge and detectability both types of polymeric compounds were derivatized with phthalic anhydride (PhAH) or 1,2,4-benzenetricarboxylic anhydride (BTA) before the separation. Derivatization with BTA yielded more complex electropherograms, due to the occurrence of different isomeric reaction products for every PEG or PPG species. Effective mobilities of the PhAH derivatives were related to the number of monomer units in the polymers in a straightforward way. The CZE method could also be used to determine the monomer-number distribution of random and block PEG-PPG copolymers. For an MEKC analysis the PEGs and PPGs were derivatized with phenyl isocyanate. Oligomers of PEGs could be separated up to molecular masses of 5,000, while for the more hydrophobic PPGs oligomeric separation was only accomplished for molecular masses of up to 1,500. Due to a strongly different separation mechanism for the PEG and PPG derivatives in the MEKC system. a complete group separation of the two types of polymer molecules could be obtained. PMID- 12580518 TI - Separation of opiate alkaloids by electrokinetic chromatography with sulfated cyclodextrin as a pseudo-stationary phase. AB - The separation of six related opiate alkaloids (morphine, thebaine, 10 hydroxythebaine, codeine, oripavine and laudanine) was studied using sulfated cyclodextrin (s-CD) as a cation-exchange pseudo-stationary phase. Cation-exchange interactions between the cationic analytes and the anionic s-CD (7-11 mol of sulfate groups per mole CD) were found to bethe predominant mechanism, allowing the separations to be performed at low pH where the opiates are protonated and exhibit very similar mobilities. The concentrations of the s-CD and the competing ion (Na+ or Mg2+) in the electrolyte were used to govern the extent of the ion exchange interactions. Interactions with the sulfated-cyclodextrin differed for each analyte, with oripavine exhibiting the strongest interaction and 10-thebaine and laudanine showing the weakest interactions. Despite the very similar structures of the analytes, these differences resulted in significant changes in separation selectivity. The separation was modelled using a migration equation derived from first principles and based on ion-exchange interactions between the s-CD and the opiates. Constants within the model were obtained by non-linear regression using a small subset of experimentally determined migration times. These constants related to the ion-exchange affinities of the s-CD for the various opiates. When the model was used to predict migration times under other experimental conditions, a very good correlation was obtained between observed and predicted mobilities (r2=0.996). Optimisation of the system was performed using the normalised resolution product and minimum resolution criteria and this process provided two optimised separations, each exhibiting a different separation selectivity. PMID- 12580519 TI - Determination of nitrosamines in preserved sausages by solid-phase extraction micellar electrokinetic chromatography. AB - A new methodology for extraction, pre-concentration and analysis of nitrosamines in meat derived products was developed. The samples (canned sausages) were distilled in vacuum steam and the nitrosamines were pre-concentrated by solid phase extraction with active carbon. Then, micellar electrokinetic chromatography was used for the separation and determination of the different nitrosamines contained in a real sample and gas chromatography with mass spectrometry detection was used as the confirmation technique. The method allowed the determination of nitrosamine compounds at trace levels with relative standard deviation ranging from 4.0 to 22%. PMID- 12580520 TI - Determination of 8-hydroxy-2'-deoxyguanosine in untreated urine by capillary electrophoresis with UV detection. AB - A capillary electrophoresis method with UV detection was developed for the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in untreated urine samples. The calibration graph for 8-OHdG in urine is linear in the concentration range 10 500 mg/l. and the detection limit is 5 mg/l (17 microM). 8-OHdG was determined in urine from oncological patients treated by radiation therapy. Its concentrations relative to creatinine were found to be in the range 10-47 microg 8-OHdG/l mg creatinine (4-19 micromol 8-OHdG/mmol creatinine). The overall time of the analysis of a urine sample was less than 15 min. PMID- 12580521 TI - Hyperlayer hollow-fiber flow field-flow fractionation of cells. AB - Interest in low-cost, analytical-scale, highly efficient and sensitive separation methods for cells, among which bacteria, is increasing. Particle separation in hollow-fiber flow field-flow fractionation (HF FlFFF) has been recently improved by the optimization of the HF FIFFF channel design. The intrinsic simplicity and low cost of this HF FlFFF channel allows for its disposable usage. which is particularly appealing for analytical bio-applications. Here, for the first time, we present a feasibility study on high-performance, hyperlayer HF FIFFF of micrometer-sized bacteria (Escherichia coli) and of different types of cells (human red blood cells, wine-making yeast from Saccharomyces cerevisiae). Fractionation performance is shown to be at least comparable to that obtained with conventional, flat-channel hyperlayer FIFFF of cells, at superior size-based selectivity and reduced analysis time. PMID- 12580522 TI - Adduct formation in electrospray ionization mass spectrometry II. Benzoic acid derivatives. AB - This work serves as a follow-up to Part I of experiments designed to determine the underlying principles in the formation of pseudomolecular, or adduct, ions during electrospray ionization. Aromatic acids were studied by flow injection analysis in the negative ionization mode of electrospray ionization mass spectrometry. Part I dealt with common acidic anti-inflammatory pharmaceuticals. such as ibuprofen and related analogues. Part II deals with functionally less complex molecules, namely benzoic acid (BA) and substituted benzoic acids. Halide substituted molecules are investigated to deduce the effect of electron withdrawing substituents (bromo-, chloro-, and fluoro-) and ring position (ortho , meta- and para-) on the response of a traditional deprotonated molecular ion ([M-H]-) and a sodium-bridged dimer ion ([2M-2 H+Na]-). Amino-substituted benzoic acids are also analyzed in order to study the effect of an additional ionizable group on the molecule, and para-tert.-butyl-BA is analyzed to study the effect of increased hydrophobicity, as they relate to the formation of pseudomolecular ions. This study shows that solution character [octanol-water partition coefficient (or log P) and pKa] of the model compounds controls the relative efficiency of formation of [M-H]- and [2M-2H+Na]- ions. However the relative gas phase character (gas phase basicity and proton affinity) also has a significant effect on the formation of the sodium-bridged dimer ion. For the halide substituted species, placement of the electron-withdrawing atom at the meta position gives the greatest enhancement in sensitivity. Observations also show that as the structural complexity of the model compound increases, predictions relating analyte acidity to sodium-bridged dimer ion formation give way to a stronger dependence between log P values and ionization efficiency. Supporting this hypothesis is the nearly ten-fold enhancement in signal for tert.-butyl BA relative to BA. due to the greater hydrophobicity, and consequently, increased surface activity in an electrosprayed droplet of the analyte molecule. PMID- 12580523 TI - A study of human skin mechanical properties by means of Cutometer. AB - Cutometer is a commercially available non-invasive suction skin elasticity meter. The present study discusses some aspects of the biological informativeness and interpretation of the results obtained in studying skin mechanical properties with cutometer. We analyze the results of previously published studies on the mechanical properties of healthy and diseased skins and their changes after external influences. Studying human skin using cutometer gives objective and biologically meaningful information about the mechanical properties of healthy and diseased human skin. PMID- 12580524 TI - A study on the basic epidemiological parameters of viral hepatitis a in the region of Plovdiv, Bulgaria. AB - INTRODUCTION: Infection with hepatitis A virus is of great importance to public health. The disease has a worldwide occurrence due to inadequate sanitation and the variable seroprevalence rates in the community. OBJECTIVE: The present report investigates the epidemiological features of hepatitis A in the region of Plovdiv and outlines the most important epidemiological features of the disease. MATERIALS AND METHODS: In 1999 (a year with a lower hepatitis A prevalence) the basic epidemiological characteristics of the disease were studied: overall morbidity and mortality rates, age, sexual and seasonal distribution, and prevalence in rural and urban areas. The study was carried out in a cohort of 288 patients with hepatitis A admitted to the Clinic of Infectious Disease of the Medical University in Plovdiv. All patients were anti HAV IgM positive. Routine clinical, laboratory, epidemiological and statistical methods were used. Laboratory and serological tests were performed at the Center for Disease Control, Plovdiv and at the University Clinical Laboratory. RESULTS: Of 423 patients with acute viral hepatitis 288 (68.09%) had hepatitis A. In 1999 the morbidity rate for the region of Plovdiv was 49.55 per hundred thousand (intermediate). Disease prevalence was the highest in preschool and early school age as well as in the age groups 20-29 (84.67 per hundred thousand) and 30-39 (71.59 per hundred thousand). Urban residents accounted for 79.85% of the cases. The male/female ratio was 1:1. 82.25% of the preschool children did not attend day-care centers. History of contact with sources of infection was elicited from 17.01% of the patients. Disease rates peaked in October through December (maximum in October). CONCLUSIONS: Accumulated data could contribute to more effective disease control. PMID- 12580525 TI - Amaryl (glimepiride) in patients with type 2 diabetes mellitus. AB - The purpose of the present study was to investigate the effect of the first third generation sulphonylurea drug glimepiride (Amaryl, Aventis) in the treatment of patients with type 2 diabetes mellitus in an open 6-month clinical trial. The study included 19 patients with type 2 diabetes mellitus (7 men and 12 women, aged 53.6 +/- 2.43 years, mean duration of diabetes 7.79 +/- 1.45 years). The body mass index (BMI) of the patients was x = 30.157 +/- 1.63 which is at the borderline between overweight and obesity. The patients started at a baseline dosage of 1 mg which was then it was gradually adjusted according to the blood sugar level. The dosage of the drug varied between 1 and 6 mg (mean daily dosage 2.03 mg). The metabolic control parameters that were calculated included fasting and 2-hour postprandial blood sugar concentration, total cholesterol, serum triglycerides, HbA1c, and microproteinuria. They were measured at baseline, at 3 and 6 months. The results showed that the fasting blood glucose decreased significantly (P<0.05 at 3 months and P<0.001 at 6 months). Statistically significant lower postprandial glycemia was also observed in the patients (the decrease was not significant at 3 months but highly significant at 6 months, P<0.01). The overall evaluation was based on the values of HbA1c--they were statistically significantly lower at 6 months (P<0.01) which suggests the steady improving tendency of the metabolic control in type 2 diabetes patients treated with Amaryl (glimepiride). The improvement of the metabolic control was also manifested by the lower serum triglycerides levels (P<0.05) and the BMI remaining nearly without change. It is concluded that Amaryl (glimepiride) is an efficacious oral sulphonylurea preparation which can be used as an appropriate substitute of the other beta cell stimulators. Glimepiride once daily provides a good compliance of patients which reduces to minimum the skipped doses. It is associated with a reduced risk of hypoglycemia and causes no weight gain. PMID- 12580526 TI - Effects of Aronia melanocarpa juice as part of the dietary regimen in patients with diabetes mellitus. AB - The low calorie juice Aronia melanocarpa (sugar free, with artificial sweeteners) could be a valuable adjunct to the complex therapy of patients with diabetes mellitus. In this study no increased blood glucose levels were established 60 min. following ingestion of 200 ml Aronia juice. On the contrary, lower fasting blood glucose concentrations were measured in 16 patients with insulin dependent diabetes and in 25 patients with non-insulin dependent diabetes (25 women and 16 men, 3 to 62 years of age, median age 38.8 +/- 4.7) with duration of the disease from 1 month to 13 years. Serial blood glucose measurements showed: 14.23 +/- 1.32 mmol/l at baseline and 11.4 +/- 0.89 mmol/l blood glucose level after 60 min., the difference being statistically significant (p<0.05). The ingestion of 200 ml Aronia juice combined with a standard breakfast produced similar results (the basal concentration of glucose was 13.43 +/- 1.12 mmol/l; it decreased to 11.94 +/- 1.02 mmol/l at 60 min., the difference did not reach statistical significance. The daily intake of 200 ml Aronia juice over a period of 3 months was effective in lowering fasting blood glucose levels from 13.28 +/- 4.55 mmol/l to 9.10 +/- 3.05 mmol/l (p<0.001) in 21 patients with non-insulin dependent diabetes--13 women and 8 men aged from 42 to 62 (median age 53.6 +/- 3.65) with disease duration from 6 to 17 years. Aronia had a beneficial effect on HbA1c, total cholesterol and lipid levels. They dropped from 9.39 +/- 2.16% to 7.49 +/- 1.33% (p<0.001), from 6.45 +/- 1.59 mmol/l to 5.05 +/- 0.96 mmol/l (p<0.001) and from 2.92 +/- 2.15 mmol/l to 1.7 +/- 1.07 mmol/l (p<0.001), respectively. Results were compared with those obtained in 23 patients with non-insulin dependent diabetes (15 women and 8 men aged from 48 to 67 years, median age 54.9 +/- 3.34) with disease duration from 6 to 17 years. The above mentioned parameters remained unchanged in these patients. Accumulated data illustrated the hypoglycemic potential of Aronia juice. The precise mechanism of its action is unknown but its beneficial effects and good taste make it a valuable adjunct to the dietary treatment of patients with diabetes mellitus. PMID- 12580527 TI - Immunoreactivity for ferritin in Leydig cells of human testis. AB - The aim of the present study was to demonstrate the immunocytochemical localization of ferritin in human Leydig cells. Testes from patients orchidectomized for carcinoma of the prostate were used. The immunoreactivity for ferritin was visualized in the Leydig cells by amplification immunocytochemical technique. The Sertoli cells and some of the germ cells show moderate immunoreactivity for the examined antigen. Our result represent an immunocytochemical verification for presence of ferritin in the human Leydig cells (also in other cellular components) and suggest the role of this factor in the local auto- and/or paracrine control of the testicular functions. PMID- 12580528 TI - Immunolocalization of neurotrophic factors and their receptors in the Leydig cells of rat during postnatal development. AB - The aim of the present study was to localize the neurotrophic factors glial cell line-derived neurotrophic factor (GDNF), nerve growth factor receptor (NGFr) and low sensitive receptor for NGF (p75LNGFR) in rat testes during the postnatal development. Rat testes obtained at different stages of the postnatal development -day 5, 10, 15, 20, 24 and 27 after birth were used. Amplification immunocytochemical technique, which includes a combination of the peroxidase antiperoxidase method (PAP) and the avidin-biotin peroxidase complex (ABC) methods, was applied. Immunoreactivity for the tested antigens was established in the Leydig cell with characteristic fluctuation in the intensity of the immune reaction at the different stages of the postnatal development. Positive immunostaining was seen in the Sertoli and some of the germ cells (spermatocytes and spermatids). The results obtained show that neurotrophic factors play a role in the processes of postnatal differentiation of the Leydig cells and in the regulation of their functional activity. PMID- 12580529 TI - Cytochemical study of in situ differentiation of white adipose cells in rat. AB - AIM: The aim of the present study was to follow up the histochemical and enzyme histochemical characteristics of differentiating white adipose cells of rat in situ. METHOD: Nine rat fetuses aged 15-21 day of gestation were used. Fragments from the subcutaneous tissue of lower limb were frozen at -18 degrees C. On consecutive cryostat sections Sudan III-hematoxylin staining and enzyme histochemical reactions for NADH2-cytochrome C-reductase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase and lipoprotein lipase were performed. RESULTS: The onset of the rat adipocyte differentiation was detected at days 15 16 of gestation. It was seen as appearance and accumulation of lipid droplets, rounding of the cells and displacement of the nuclei to the cell periphery. Gradually, with the advance of gestational age the number of differentiating adipose cells increased and lipid packages, composed of mature-like unilocular adipocytes and multilocular adipocytes that had not yet completed their differentiation, were formed. The differentiating embryonal adipose cells expressed positive enzyme-histochemical reactions for NADH2-cytochrome C reductase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase and lipoprotein lipase. CONCLUSION: The described histochemical and enzyme histochemical characteristics could be used as markers for distinguishing the earliest embryonal adipose cells in rat, which is impossible with the classic histological techniques. At the same time they might be accepted as the primary morphological criteria of adipocyte differentiation in situ. PMID- 12580531 TI - An in vitro study of the released potassium and chloride ions from polycarboxylate cement containing 5% KCl. AB - AIM: To find with a laboratory study in vitro the concentrations of released potassium (K+) and chloride (Cl-) ions extracted from polycarboxylate cement containing 5% KCl. MATERIAL AND METHODS: Model calibrated blocks of polycarboxylate cement containing 5% KCl were prepared for the study. Every block contained 6.40 mg potassium ions and 5.80 mg chloride ions. The extraction of the ions was performed in stationary conditions with extragent Aqua redestillata at different time periods: 20 minutes from the mixing of the cement, 24 h, 7 days/168 h/, 14 days/336 h/, 30 days/720 h/ and 90 days/2160 h/ at temperature 36 +/- 0.5 degrees C and pH--7. The concentrations of the released K- and Cl- ions in the extract (in mg/10 cm3), as well as their degree of extraction (in percentage) were measured using emission flame photometry and spectrophotometry. The data are analyzed statistically using descriptive statistics, statistical significance being defined at p<0.05. RESULTS: The concentrations of released potassium and chloride ions increased with time and reached their maximum on the 14th day (336 h). The differences with their respective baseline values are statistically significant (P<0.001). The degree of extraction of the active ions is highest on the 14th day (336 h) - (93.12% for the K-ions and 93.10% for the CL ions). CONCLUSION: The active supplement of 5% KCL in the polycarboxylate cement increases the rate of extraction of the ions, as the concentrations of the released K- and Cl-ions increase with time and reach maximum values on the 14th day (336 h). PMID- 12580530 TI - Dexamethasone and oxytocin effects on rat Leydig cells--morphological and enzyme histochemical characteristics. AB - In the present study the fine structure of rat Leydig cells was examined by electron microscopy. Oxytocin and dexamethasone induced changes in the activities of 3beta hydroxysteroid dehydrogenase, NADH2 cytochrome-C-reductase and glucose-6 phosphate dehydrogenase in these cells were studied in an in vivo experiment. Two groups of male Wistar rats were used to test the effects of oxytocin--rats from group I received a single injection of oxytocin; rats from group II were given a 10-day course of oxytocin. Pregnant female rats were injected with dexamethasone on day 17 and 18 post conception. The testes of 19 and 20-day old embryos were removed. It was established that both short and long term courses of oxytocin increased the activities of the above-mentioned enzymes. On the contrary, prenatal administration of dexamethasone decreased enzymatic activity in Leydig cells. Electron microscopy revealed clusters of fetal Leydig cells. Our results indicated the role of oxytocin in the local regulation of steroidogenesis and the importance of glucocorticoids in the differentiation and activity of Leydig cells. PMID- 12580532 TI - Qualities of the restorations of glass-ionomer cement of caries and non-caries lesions of the dental necks after a two-year observation period. AB - AIM: The aim of the study was to follow up the quality change of the restorations with the glass-ionomer cement "Vitremer" 3M. MATERIAL AND METHODS: A total of 97 restorations were performed in the region of the dental necks. The diagnoses were as follows: caries cervicalis--55; caries radicis dentis--9; usura cervicalis--9; erosio dentis--24. The assessment of the restorations was performed immediately after fixing and two years later according to the criteria of G Ryge. RESULTS: The results showed good adhesion of the restorative material to the dental tissues, absence of secondary caries and changes in the marginal adaptation and considerable darkening of the colour. CONCLUSION: The glass-ionomer cement "Vitremer" 3M is suitable restorative material for the cervical region, especially in individuals exhibiting high caries activity. PMID- 12580533 TI - Mechanism of cartilage destruction in osteoarthritis. AB - Osteoarthritis (OA) is one of the most common diseases among the elderly. For many years, OA was considered a normal result of the aging process with few treatment options. Remarkable progress in understanding OA cartilage has been achieved in recent years. The application of technical advances to clinical studies of chondrocytes and cartilage tissue metabolism will provide important new insights concerning the pathophysiology of OA and identify new therapeutic strategies to regulate and inhibit the degenerative process of OA. However, many problems remain unresolved. In this review we try to focus on recent advances in the field of cartilage metabolism and molecular markers to facilitate the determination of a patients prognosis and the need for cartilage protective treatment. PMID- 12580534 TI - Functional activation of peroxisome proliferator-activated receptor alpha (PPARalpha) by environmental chemicals in relation to their toxicities. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) may work in the processes of both physiological and toxicological response to various endogenous or exogenous substances. The literature on the study of functional activation of PPARalpha by environmental chemicals in relation to their toxicities were reviewed. Environmental chemicals that were found to induce peroxisomes (peroxisome proliferators) and to activate the function of PPARalpha included plasticizers, herbicides, and organic solvents that have carboxyl groups in their parent substances or their metabolites. Several studies have showed species differences in the constitutive expression of PPARalpha and activation of PPARalpha, which may result in species differences in the induction of transcription of the genes encoding several peroxisomal enzymes. Although much information has supported the view that PPARalpha is primarily involved in the hepatic carcinogenicity of peroxisome proliferators, conflicting evidence exists. Most of the peroxisome proliferators have been shown to induce reproductive and developmental disorders, which might, in part. be associated with the functional activation of PPARalpha. Few epidemiological studies on the effect of peroxisome proliferators on humans have been conducted. The effect of perfluorooctanoic acid on humans was evaluated from the aspect of lipid metabolism in one study, which concluded that there was no effect. PMID- 12580535 TI - Simple exponential regression model to describe the relation between minute ventilation and oxygen uptake during incremental exercise. AB - The physiological significance of an exponential regression model between minute ventilation (VE) and oxygen uptake (VO2) during incremental exercise was examined. Thirty-eight subjects, including 12 patients with chronic heart failure, participated in cardiopulmonary exercise testing on a bicycle ergometer. The equation VE = a e(bVO2), where a and b are parameters, was used to describe the relation between VE and VO2 during incremental exercise. Arterialized blood gas analysis was measured before and during exercise. The correlation coefficient of the regression model was high (r = 0.97 +/- 0.02). Parameter a negatively correlated with the arterial partial pressure of carbon dioxide during exercise (r = -0.44, p < 0.01), and positively correlated with peak VO2 (r = 0.47, p < 0.01). Parameter b negatively correlated with peak VO2 (r = -0.86, p < 0.01) and positively correlated with the dead space to tidal volume ratio (r = 0.68, p < 0.01). The regression model, as well as parameters a and b, is physiologically useful in expressing metabolic response to exercise. This model, a specific solution to the differential equation dVE/dVO2 = bVE, implies that the more a subject breathes, the greater is the increment in ventilation needed to meet a further increment of metabolic demand. PMID- 12580536 TI - Total hip arthroplasty for a patient with angel-shaped phalango-epiphyseal dysplasia (ASPED). A case report. AB - Angel-shaped phalango-epiphyseal dysplasia is characterized by the angel shape of the middle phalanx and severe coxarthrosis in adult life. This osteochondrodysplasia shows a further variety of heterogeneous multiple epiphyseal dysplasias. It also shows a late and dysplastic development of the femoral head that leads to osteoarthrotic changes with severe hip pain and gait disturbance. In this report, we show a 35-year-old female with Angel-shaped phalango-epiphyseal dysplasia that was treated by bilateral total hip arthroplasty. She has suffered from her coxalgia since she was 27 and since the age of 30 has been able to walk with the aid of crutches. The radiographs of her bilateral hip showed severe osteochondrotic changes with a progressive disappearance of the joint space. Total hip arthroplasties were performed to treat the osteoarthrosis on the bilateral hip joint due to Angel-shaped phalango epiphyseal dysplasia. One year after surgery, she is pain-free, and able to walk with a cane. Based on this case, we propose that total hip arthroplasty should be considered one of the treatments for the coxopathy in patients with Angel-shaped phalangoepiphyseal dysplasia. PMID- 12580537 TI - Effects of moderate exercise on metabolic responses and respiratory exchange ratio (RER). AB - To counteract insulin resistance, it is necessary to increase the utilization rate of fatty acids in blood and adipose tissue. The aim of the present study was to determine the relation between metabolic responses and exercise duration from changes in the respiratory exchange ratio (RER). The mean RER during 30 minutes of moderate exercise (mean pulse 115 beats/min) was 0.89 +/- 0.02, indicating no major change. Significant changes were observed in the levels of plasma glucose (PG), plasma free fatty acid (FFA), and plasma immunoreactive insulin (IRI) before and after exercise, demonstrating a decrease in PG and IRI, and an increase in FFA levels. However, the RER value indicated that carbohydrate was the dominant metabolic substrate; therefore, prolonged or repetitive brief and mild to moderate exercise is necessary to increase the utilization of fatty acids. PMID- 12580538 TI - Loneliness as expressed by schizophrenic patients in the early remission phase. AB - We examine the clinical and psychotherapeutic significance of the "emotion of depression," particularly "loneliness," focusing on the postpsychotic depression in one phase of the early remission phase and prolonged early remission phase according to the so-called "remission process theory" of schizophrenia (Nakai). We first present details of two cases representative among 30 patients who discussed "loneliness" with their chief therapist. Then, in the Discussion, we classify the expressions of "loneliness" into four modes, taking "loneliness as isolation open to the other." In this phase, the patient strongly seeks a "deeply significant other" as a "partner for even a little protection against loneliness," and the therapist often assumes this role. This role is extremely important to the patient's passage through the early remission phase. We stress that the presence of this "partner for even a little protection against loneliness" is of great significance for the problems of subsequent progress or stagnation in the remission process, and the issue of ongoing deterioration as well. PMID- 12580539 TI - Statistical method in a comparative study in which the standard treatment is superior to others. AB - The statistical method in a comparative study in which the standard treatment is theoretically or practically superior to the others has been investigated for a matched-pairs design. We derived this statistical method from one based on the maximum likelihood and score methods. Here we have shown that the score test statistic is algebraically the same as the statistic from the maximum likelihood method. As an example of our method's applications, we have considered a study on the detection of nodules on chest X-ray images displayed on a CRT with low luminance. PMID- 12580540 TI - Upregulation of group 1 CD1 antigen presenting molecules in guinea pigs with experimental autoimmune encephalomyelitis: an immunohistochemical study. AB - In humans, group 1 CD1 glycoproteins present foreign and self lipid and glycolipid antigens to T-cells. Homologues of these molecules are not found in mice or rats but are present in guinea pigs (GPs). We examined CD1 and MHC class II expression in the central nervous system (CNS) of GPs sensitized for experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. In normal GPs and the uninflamed CNS, low-level MHC class II (MHC II) immunoreactivity occurred on vascular elements, meningeal macrophages and parenchymal microglial cells, whereas immunoreactivity for CD1 was absent. In the inflamed CNS, the majority of infiltrating cells were MHC II+ and microglia showed increased expression. CD1 immunoreactivity was detected on astrocytes and subsets of inflammatory cells Including B cells and macrophages. Minimal CD1 and MHC II co-expression was noted on inflammatory cells or glia. We conclude that group 1 CD1 molecules are strongly upregulated in the inflamed CNS on subsets of cells distinct from the majority of MHC II bearing cells. The expression of CD1 proteins in such lesions broadens the potential repertoire of antigens recognized at these sites and highlights the value of the GP as a model for studies of the relevance of CD1 molecules in host defense and autoimmune diseases. PMID- 12580541 TI - Neuropathology with clinical correlations of sporadic amyotrophic lateral sclerosis: 102 autopsy cases examined between 1962 and 2000. AB - Sporadic amyotrophic lateral sclerosis (ALS) is a fatal neurological disorder affecting adults. We studied the neuropathology and clinical correlations in 102 autopsy cases of ALS. The age at onset of the disease was significantly higher for the bulbaronset form (30 cases) than for the limb-onset form (72 cases). Dementia was confirmed in 7 cases. These 102 cases were divided into 4 pathological subgroups: typical ALS (59 cases), lower-motor-predominant ALS (23 cases), ALS with temporal lesions (18 cases), and ALS with pallido-nigro-luysian degeneration (2 cases). The age at onset was significantly higher for lower-motor predominant ALS and ALS with temporal lesions than for typical ALS. In the lower motor neurons, Bunina bodies were detected in 88 cases, whereas ubiquitin immunoreactive skein and/or spherical inclusions were detected in all 102 cases. Of the 100 available cases, 50 and 16 also showed ubiquitin-immunoreactive inclusions in the neostriatal and temporal small neurons, respectively. Ubiquitin immunoreactive dystrophic neurites were also observed in the neostriatum in 3 of the 50 cases with neostriatal inclusions, and in the temporal cortex in 4 of the 16 cases with temporal inclusions. There was a significant association between the bulbar-onset form, temporal lesions, neostriatal inclusions and temporal inclusions, and between dementia, temporal lesions and temporal inclusions. Neostriatal and temporal dystrophic neurites were associated with dementia and bulbar-onset form through temporal lesions and temporal inclusions. The present findings may be helpful for designing further studies on the mechanisms underlying the development of ALS. PMID- 12580542 TI - Inflammation in the central nervous system: the role for dendritic cells. AB - Dendritic cells (DCs) are a subclass of antigen-presenting cells critical in the initiation and regulation of adaptive immunity against pathogens and tumors, as well as in the triggering of autoimmunity. Recent studies have provided important knowledge regarding distribution of DCs in the central nervous system (CNS) and their role in intrathecal immune responses. DCs are present in normal meninges, choroid plexus, and cerebrospinal fluid, but absent from the normal brain parenchyma. Inflammation is accompanied by recruitment and/or development of DCs in the affected brain tissue. DCs present in different compartments of the CNS are likely to play a role in the defence against CNS infections, and also may contribute to relapses/chronicity of CNS inflammation and to break-down of tolerance to CNS autoantigens. CNS DCs can therefore be viewed as a future therapeutic target in chronic inflammatory diseases such as multiple sclerosis. PMID- 12580543 TI - Green fluorescent protein immunohistochemistry as a novel experimental tool for the detection of glioma cell invasion in vivo. AB - In vivo animal models of primary brain tumors are necessary to advance knowledge related to the complex interactions between glioma cells and the adjacent brain. A cardinal feature of glioma growth, and a major reason why neurosurgical and adjunctive therapies ultimately fail in most patients is their invasive properties. We have adapted a previously described animal model developed by one of us to give better histological detail while preserving the identification of single infiltrating glioma cells. GL261 glioma cells were first transfected with the plasmid encoding green fluorescent protein (GFP) and then implanted into the brains of syngeneic C57BL/6 mice. Identification of GFP-positive tumor cells in paraffin sections of the brains of tumor-bearing animals utilized an antibody for conventional immunoperoxidase immunohistochemistry. This method is a more powerful technique compared with the prior use of frozen sections and fluorescence microscopy to identify GFP-tagged tumor cells. We find that this new method provides improved morphology and proves to be a sensitive and reliable system for detection of invading glioma cells. Using this methodology with other advanced technologies (eg, laser capture microdissection) holds out the promise of helping to elucidate the molecular mechanisms of glioma cell infiltration and invasion into the surrounding brain. PMID- 12580544 TI - Lymphoid chemokines CCL19 and CCL21 are expressed in the central nervous system during experimental autoimmune encephalomyelitis: implications for the maintenance of chronic neuroinflammation. AB - The simultaneous presence of dendritic, T- and B-cells in the central nervous system (CNS) of mice with experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, suggests that interactions among these cell types might be instrumental in the local induction and maintenance of autoimmune reactions. In this study, we explored the possibility that such aberrant leukocyte recruitment in the CNS could be sustained by "lymphoid" chemokines which orchestrate dendritic cell and lymphocyte homing to lymphoid organs. Transcripts for CCL19 and CCL21 and their common receptor CCR7 were induced in the CNS of mice undergoing relapsing-remitting and chronic-relapsing EAE. While CCL21 immunoreactivity was confined to the endothelium of some inflamed blood vessels, CCL19 was expressed by many infiltrating leukocytes and some astrocytes and microglia in the CNS parenchyma. CCR7+ cells accumulated in inflammatory lesions during EAE progression, when abundant infiltration of the CNS by mature dendritic cells, B-cells and cells expressing naive T-cell markers also occurred. These findings suggest that CCL19 and CCL21 produced in the EAE-affected CNS may be critical for the homing of antigen presenting cells and lymphocytes, resulting in continuous local antigenic stimulation and maintenance of chronic neuroinflammation. PMID- 12580545 TI - Targeted molecular therapy of GBM. AB - Major advances in molecular biology, cellular biology and genomics have substantially improved our understanding of cancer. Now, these advances are being translated into therapy. Targeted therapy directed at specific molecular alterations is already creating a shift in the treatment of cancer patients. Glioblastoma (GBM), the most common brain cancer of adults, is highly suited for this new approach. GBMs commonly overexpress the oncogenes EGFR and PDGFR, and contain mutations and deletions of tumor suppressor genes PTEN and TP53. Some of these alterations lead to activation of the P13K/Akt and Ras/MAPK pathways, which provide targets for therapy. In this paper, we review the ways in which molecular therapies are being applied to GBM patients, and describe the tools of these approaches: pathway inhibitors, monoclonal antibodies and oncolytic viruses. We describe strategies to: i) target EGFR, its ligand-independent variant EGFRvIII, and PDGFR on the cell surface, ii) inhibit constitutively activate RAS/MAPK and PI3K/Akt signaling pathways, iii) target TP53 mutant tumors, and iv) block GBM angiogenesis and invasion. These new approaches are likely to revolutionize the treatment of GBM patients. They will also present new challenges and opportunities for neuropathology. PMID- 12580546 TI - Phosphorylated protein kinases associated with neuronal and glial tau deposits in argyrophilic grain disease. AB - Tau phosphorylation was examined in argyrophilic grain disease (AGD) by using the phosphospecific tau antibodies Thr181, Ser202, Ser214, Ser 396 and Ser422, and antibodies to non-phosphorylated and phosphorylated mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinases (ERK), stress-activated kinase (SAPK), c-Jun N-terminal kinase (JNK), p38 kinase (p-38), alpha calcium/calmodulin-dependent kinase II (alphaCaM kinase II), and glycogen synthase kinase-3 (GSK-3), all of which regulate phosphorylation at specific sites of tau. This is the first study in which the role of protein kinases in tau phosphorylation has been examined in AGD. Hyperphosphorylated tau accumulated in grains and pre-tangles in the hippocampus, dentate gyrus, entorhinal and trans entorhinal cortices, and amygdala in all cases. Ballooned neurons in the amygdala, entorhinal, insular and cingulate cortex, and claustrum contained alphaB-crystallyn and phosphorylated neurofilament epitopes. Some astrocytes and scattered oligodendrocytes containing coiled bodies were recognized with anti-tau antibodies. A few tangles were observed in the entorhinal cortex and hippocampus corresponding to Alzheimer's disease (AD) stages I-III of Braak and Braak. None of the present cases was associated with progressive supranuclear palsy or with alpha-synuclein pathology. Two bands of phospho-tau of 64 and 68 kDa were observed in Western blots of sarkosyl-insoluble fractions enriched with abnormal filaments in AGD, a pattern that contrasts with the 4-band pattern obtained in AD. No modifications in the expression of non-phosphorylated MEK-1, ERK2 and GSK 3alpha/beta, as revealed by immunohistochemistry, were seen in AGD, but sarkosyl insoluble fractions were particularly enriched in JNK-1 and alphaCaM kinase II. Increased expression of the phosphorylated (P) forms of MAPK/ERK, SAPK/JNK, p38 and GSK-3beta was found in grains and tau-containing cells in AGD. MAPK/ERK-P immunoreactivity was observed in pre-tangles and, diffusely, in the cytoplasm of ballooned neurons, but not in grains. Strong SAPK/JNK-P and P38-P, and moderate GSK-3b-P immunoreactivities selectively occured in grains, in neurons with pre tangles and in the peripheral region of the cytoplasm of ballooned neurons. MAPK/ERK-P, SAPK/JNK-P, p38-P and GSK-3beta-P were expressed in tau-containing astrocytes and in oligodendrocytes with coiled bodies. Western blots revealed kinase expression in sarkosyl-insoluble fractions but none of the phospho-kinase antibodies recognized hyper-phosphorylated tau protein. These findings indicate complex, specific profiles of tau phosphorylation and concomitant activation of precise kinases that have the capacity to phosphorylate tau at specific sites in AGD. These kinases co-localize abnormal tau in selected structures and cells, including neurons with pre-tangles, ballooned neurons, astrocytes and oligodendrocytes. Most of these kinases are involved in cell death and cell survival in certain experimental paradigms. However, double-labeling studies with the method of in situ end-labeling of nuclear DNA fragmentation and cleaved (active) caspase-3 immunohistochemistry show no expression of apoptosis and death markers in cells bearing phosphorylated kinases. PMID- 12580547 TI - The neuropathology of HIV infection in the era of Highly Active AntiRetroviral Therapy (HAART). PMID- 12580549 TI - HIV-1 persistence, viral reservoir, and the central nervous system in the HAART era. AB - HAART therapy has led to a significant reduction of general and neurological morbidity, and mortality among HIV-1 infected patients. It can also decrease HIV 1 RNA titres in plasma and CSF towards undetectable level. However, the initial hope of achieving total eradication of the virus from the body has vanished. Even in patients who do not develop viral resistance or treatment intolerance, two kinds of viral persistence have been demonstrated both in lymphoid and central nervous system. The first one is a smoldering infection that persists, despite prolonged and apparently efficient HAART, in monocytes, tissue macrophages and most probably microglia. The second one is an integration of proviral DNA in the genome of subpopulations of CD4 lymphocytes of patients receiving efficient HAART. A similar viral integration in astrocytes and less likely in resting microglia is suggested by several studies, although it has yet to be demonstrated conclusively. PMID- 12580548 TI - Astrocyte activation and apoptosis: their roles in the neuropathology of HIV infection. AB - Astrogliosis is a common neuropathological finding in the brains of HIV infected individuals; both activation and apoptosis of astrocytes are seen. This review aims to discuss the Fas pathway in the context of proliferation and apoptosis of astrocytes during HIV infection, and as a result of astrogliosis, the dysregulation of astrocyte-neuron networks. The presence of molecules reflecting astrocyte activation, which are derived from the solubilization of receptor/ligand from the surface of proliferating astrocytes, in the cerebrospinal fluid may be used to evaluate the degree of brain cell activation during HAART therapy. A better understanding of the molecular pathway(s) leading to increase activation and apoptosis of astrocytes, in parallel with studies conducted to unravel the molecules involved in T-cell apoptosis during HIV infection, may lead to the development of new therapeutic strategies for controlling HIV replication and tissue damage. PMID- 12580550 TI - Assessing the efficacy of highly active antiretroviral therapy in the brain. AB - The devastating effects of HIV infection have been documented for the last 2 decades. Since the 1980s over 60 million people have been infected and at present 40 million people globally are living with HIV. HIV infects the central nervous system (CNS) early in the disease process. Indeed, numerous studies document the presence of HIV within the cerebrospinal fluid (CSF). Direct infection of the brain by HIV ultimately results in HIV associated dementia (HAD), which (prior to the advent of antiretroviral therapy) affected 20% of patients. An increasing number of drugs have been developed to treat this infection and delay the development of AIDS. Current treatment is aimed at inhibiting viral replication, and thus, lowering the viral load. However a subsequent increase in viral load can occur as patients become resistant to drug therapy. In the era of HAART, the incidence of HAD has been reduced, whereas the prevalence rate is increasing as people with HIV survive longer. However, in a study of initial AIDS defining illnesses, the proportion with HIV related dementia did not decline following introduction of HAART. In a separate study, no decrease was found in the incidence of dementia per se, although there was a decrease in the incidence of all AIDS-defining illnesses during this time period. It is evident from most studies that since the introduction of HAART, its effect on HAD is not entirely clear, although the majority of findings indicate that it is beneficial. Here we will outline the issues relevant to preventing HAD by HAART. PMID- 12580551 TI - July 2002: 66-year-old female with a one-year history of progressive left proptosis. AB - The July 2002 Case of the Month (COM). This 66-year-old Caucasian female presented with gradually increasing protrusion of her left eye over a one-year period. She complained of increased tearing and foreign body sensation. The physical examination revealed a visual acuity of 20/20, normal color testing, full vision field with motility of her left eye limited in lateral gaze. Pupils were round, symmetric, with no afferent pupillary defect noted. On external examination, her left eye was grossly proptotic with resistance to retropulsion. She had 4 mm proptosis of the left eye. Computed tomography and MR imaging demonstrated a left retro-orbital mass with gadolinium enhancement and focal remodeling of orbital bones. She underwent surgical resection of tumor with a diagnosis of solitary fibrous tumor, and postoperatively she was symptom free. The histopathological differential diagnoses of spindle cell neoplasms of the orbit are discussed. Five months after surgery, no evidence of tumor recurrence was seen on neuroimaging and her vision was 20/20. PMID- 12580552 TI - August 2002: 21-year-old male with cystic intracerebral tumor. AB - The August 2002 COM. A 21-year-old male presented with a single episode of generalized tonic clonic seizures. Radiology revealed a cystic tumor with mural nodule suggestive of a pilocytic astrocytoma. However, histopathological examination and electron microscopy revealed features of an intracerebral schwannoma. Therefore, although rare, in an intracerebral cystic lesion with mural nodule, the possibility of an intracerebral schwannoma should be entertained. This is important because this is a benign tumor with favourable response to resection. PMID- 12580553 TI - September 2002: 24-year-old female with a 6-month history of seizures. AB - The September 2002 COM. A 24-year-old female presented with a history of 3 generalized seizures, the first of which had occurred 6 months before admission. Her neurological examination was normal, but upon admission her MRI showed a small cystic lesion in the left parieto-occipital region. The lesion was hyper intense on T-2 weighted images and did not show contrast enhancment. At surgery, the tumor was found to be deep to the cortex and was a cyst with amber fluid surrounded by gliotic brain. Microscopically, the tumor was well-demarcated from the surrounding tissues, which showed reactive changes, including Rosenthal fibers. The tumor was composed of GFAP-positive glial cells, which were arranged in a pseudopapillary fashion around blood vessels. In between, the tumor cells were positive for neuronal markers. The diagnosis was papillary glioneuronal tumor (PGNT), a relatively recently described lesion that may be a variant of ganglioglioma. The current literature on PGNTs is reviewed. PMID- 12580554 TI - Implications of diabetes mellitus in urology. AB - The incidence of DM is rapidly growing among Americans. DM will rival cancer and heart disease in terms of cost and suffering. The National Institute of Health is tripling the research dollars that are spent on diabetic-related research in an attempt to combat this disease. Urologists are on the front line in the diagnosis and treatment of the complications of DM. The complications of DM that we reviewed in this article, diabetic cystopathy and diabetic ED, can occur in the early stage of DM and often progress in a silent fashion. More awareness and interest are needed to improve our understanding of diabetic complications in urology. Exciting new approaches in the treatment of diabetic cystopathy and ED are being investigated. PMID- 12580555 TI - Urological manifestations of vascular disease. AB - We have detailed several of the urological manifestations of vascular disease. With the aging of the North American population, urologists will encounter the urological complications of vascular disease with ever-increasing frequency. PMID- 12580556 TI - Neurological disorders cerebrovascular disease and parkinsonism. AB - To store and expel urine at appropriate intervals, the lower urinary tract requires extensive input and control from the peripheral autonomic, somatic, and central nervous systems. Neurological disorders, such as cerebrovascular disease and Parkinson's disease, often cause functional disturbances of the lower urinary tract. PMID- 12580557 TI - Urologic manifestations of hematologic disease sickle cell, leukemia, and thromboembolic disease. AB - Advances in medicine are allowing patients with hematologic disease to live longer and healthier lives than ever before. As these patients age, however, manifestations of their disease processes may develop as complications in other organ systems. We discussed the major genitourinary complications of sickle cell anemia, leukemia, and thromboembolic disease. These range from the benign inability to concentrate urine that is seen in sickle cell disease to renal infarction that results from nephrotic syndrome. Our ability to treat and prevent these complications will improve as our understanding of these disease processes and their pathophysiology grows. Additionally, it is important for urologists to understand the underlying pathophysiology of hematologic disease to best serve the patients. For example, it may be the urologist who makes the diagnosis of ovarian vein thrombosis in a pregnant woman with right lower quadrant pain and fever. This diagnosis, with the proper treatment of antibiotics and anticoagulation, could prevent the potential development of septic thrombophlebitis. Urologists will increasingly be called upon to deal with the manifestations of these complex diseases as these patients are living longer. It is our duty to educate ourselves about these disease processes so that we can make the best clinical decisions for our patients. PMID- 12580558 TI - Urologic manifestations of nonurologic diseases gastrointestinal disorders. AB - A variety of common, gastrointestinal diseases result in significant genitourinary tract pathology. In general, knowledge of these associated disease processes permit rapid and accurate diagnosis and treatment. The underlying thread is the recognition of one pathophysiological process to explain patterns of a single disease. PMID- 12580559 TI - Endocrine causes of impotence (nondiabetes). AB - The endocrine system has a major role in erections in normal men and it can also be a cause of significant morbidity. The relationship between serum testosterone measurement and erectile function is complex. Androgen treatment should certainly be considered in patients without prostate cancer but with a clinical picture that suggests a relevant contribution of hypogonadism to the ED. Other, nondiabetic, endocrine abnormalities may need to be considered in the management or the patient with ED. PMID- 12580560 TI - Endocrine causes of male infertility. AB - Although endocrinopathies are not often seen in infertile men, these disorders are clinically significant; they often have potentially serious medical significance, regardless of fertility issues. Correction of these disorders represents a possible way to restore normal fertility for the male partner. Male fertility is critically dependent upon a normal hormonal milieu. The hypothalamic pituitary-gonadal axis is quite sensitive to disruption by endocrine disorders and other generalized medical disorders. Thus, male infertility is occasionally the presenting sign for significant underlying medical disease; it is important to properly evaluate these patients. PMID- 12580561 TI - Urologic manifestations of nonurologic disease urolithiasis. AB - Patients with nephrolithiasis may have coexistent diseases that play a causative role in stone formation. A stone event may be the initial manifestation of the disorder and the urologist may play a major diagnostic role. Regulation or correction of the disorder may eradicate or dramatically attenuate stone activity. In some patients, metabolic abnormalities persist, however, and the therapeutic strategies that were reviewed in this manuscript may need to be implemented. Stone disease may also be induced by drugs that are used to treat a number of disease processes. When this occurs, the drug will usually need to be discontinued and alternative therapy should be instituted. PMID- 12580562 TI - AIDS and the urologist. AB - The management of HIV infection has dramatically altered the natural history of the disease. Prevention of opportunistic infections and the development of HAART regimens altered the manifestations and conditions that urologists are being asked to evaluate and manage in this patient population. PMID- 12580563 TI - Genitourinary manifestations of tuberculosis. AB - By the 1980s, the availability of antituberculosis chemotherapy reduced the incidence and prevalence of tuberculosis. Changing patterns of population emigration and the development of large pools of immune-compromised individuals reversed the downward trend of tuberculosis. The incidence of genitourinary tuberculosis has remained constant. The manifestations of GU TB can be variable and cause a variety of clinical patterns that mimic other diseases. Adrenal insufficiency, renal disease, obstructive uropathy, and chronic cystitis are not uncommon with TB. The patient with TB may have genital disease that simulates STD or scrotal tumors. Infertility can be caused by GU tuberculosis. Awareness of environmental factors and patient history should alert the urologist to the wide array of clinical findings in the genitourinary system that can be caused by tuberculosis. PMID- 12580564 TI - Urologic complications of nonurologic medications. AB - A physician must be aware of common drug side effects and interactions before prescribing a certain agent. In addition to the drugs that we, as urologists, prescribe, we must also be aware of the urologic side effects of drugs that are commonly prescribed by nonurologists. The mechanisms of the pharmacologic causes for voiding dysfunction, erectile and sexual dysfunction, infertility, and urolithiasis are often mutifactorial and incompletely understood. The recognition and association of a particular drug's potential side effects may save valuable time and money involved in the workup of a patient with a new urologic complaint. It is incumbent on the practicing urologist to be able to recognize the common, and sometimes subtle, urologic complications of medications that are used for nonurologic conditions. PMID- 12580565 TI - Genetic disorders and renal cell carcinoma. AB - The study of hereditary RCC syndromes continues to provide significant insight into the pathways that are involved in renal cell tumorigenesis. The clinician should maintain a high level of suspicion for genetic disorders when patients present with early-onset or mult-focal RCC. Recognition of familial syndromes will facilitate the institution of parenchymal sparing measures, as well as appropriate screening and intervention for associated nonrenal manifestations. PMID- 12580566 TI - Genetic disorders and infertility. AB - Many aspects of male fertility are influenced by genetics. Over 150 genes have been shown to be associated with infertility in mouse models, although translation of these findings to human male infertility has been slow. Nevertheless, it is likely that a significant number of these gene deletions may be associated with human infertility. There is much that we do not understand about the molecular basis of human male infertility; patients should be advised of this caveat. Genetic disorders in humans can lead to impaired spermatogenesis, defective sperm function, and defects in delivery of sperm. It is critical for the urologist who evaluates and treats infertile couples to have a working knowledge of these disorders. An understanding of the genetic basis of male infertility allows for the appropriate counseling of patients about treatment options and risks to their potential offspring. PMID- 12580567 TI - Genetic diseases in adults. AB - Genetic diseases that do not primarily affect the genitourinary tract may have urologic manifestations. These urologic manifestations range from benign and malignant renal disease to infertility. Thus, the practicing urologist may be involved in the care of these patients and should have knowledge of these diseases. Continued improvements in the diagnosis and treatment of these genetic diseases will likely result in improved survival and will increase the number of patients who may develop urologic manifestations of these diseases. PMID- 12580568 TI - Genetic diseases pediatrics. AB - The field of genetics will continue to grow because of advances in molecular biology. These advances will lead to further understanding of genetic conditions, which will enhance the urologist's knowledge of the urologic manifestations of these conditions. This will enable improved parental counseling and patient care. PMID- 12580569 TI - Stress-like responses to common procedures in individually and group-housed female rats. AB - The objective of this study was to assess the cardiovascular function and behavior of female Sprague-Dawley rats housed individually or with one or three cage mates under resting conditions and when subjected to common husbandry and experimental procedures and potentially stressful olfactory stimuli. Heart rate (HR) and mean arterial blood pressure (MAP) were assessed continuously by using radiotelemetry and are reported for the following periods: for 1 hour each day prior to any human interaction; for 12 h each day during the dark phase of the 12:12-h light: dark photoperiod; and for 2 h before and 3 h after acute husbandry and experimental procedures. Home-cage behaviors (sleeping, awake but not moving, moving, rearing, and grooming) were scored once each minute for 15 min before and 45 min after the acute procedures. Mean resting HR values in the mornings prior to human contact were significantly (P < 0.05) lower in rats housed four per cage than animals housed alone or with one cage mate, whereas MAP during this period was lowest in rats housed two per cage. Nocturnal HRs were highest in rats housed two per cage, whereas nocturnal MAP did not differ significantly between housing groups. When rats were subjected to acute husbandry and experimental procedures, HRs increased 80 to 180 beats per min (bpm) above a baseline of 300 to 325 bpm and were significantly (P < 0.05) increased for periods of 30 to 90 min after the procedures. MAP showed increases that were proportionately the same as those in HR. Group housing often, but not always, reduced these cardiovascular responses. Procedure-induced arousal behaviors occurred in all housing groups after the acute husbandry and experimental procedures, but the occurrence of these behaviors was less frequent and of shorter duration in group-housed rats than rats housed alone. In light of these results, we conclude that under resting conditions group housed rats were somewhat less stressed than were rats housed alone. Further, we conclude that common procedures induce significant stress-like responses in female rats, and the magnitude and duration of these responses are reduced by group housing. PMID- 12580570 TI - Are "by-stander" female Sprague-Dawley rats affected by experimental procedures? AB - The objective of this study was to test the hypotheses that female rats are stressed by being in the same room as animals subjected to common husbandry and experimental procedures and that the level of stress is affected by housing density. Two commonly used indices of stress, heart rate (HR) and mean arterial blood pressure (MAP), were determined by using radiotelemetry for 2 h before and 3 h after rats witnessed the following procedures: decapitation, simulated decapitation, cage change, simulated cage change, restraint and subcutaneous injection, removal of rats to another room for injection, restraint and tail-vein injection, handling and weighing, and handling and vaginal lavage. In addition, home cage behaviors (sleeping, awake, moving, rearing, and grooming) were scored once each minute for 15 min before and 45 min after the procedures. Witnessing decapitation of six other rats induced small, but significant, increases in HR above undisturbed baseline values in animals housed alone, whereas responses in animals housed with one or three cage mates were slightly greater than those of rats housed alone. Witnessing a routine cage change induced significant increases in HR in rats which were equal to or greater than those induced by witnessing decapitations; however, housing density had little effect on the responses to cage change. HR did not significantly increase above baseline values in rats witnessing restraint and a subcutaneous or tail-vein injection of other rats or when witnessing other rats being handled and weighed. However, rats housed alone showed significant increases in HR when witnessing a vaginal lavage of other rats. Active behaviors (moving, rearing, grooming) in the home cage were significantly altered only in rats housed alone and then only when witnessing a cage change or a tail-vein injection. Considering primarily increased HR, we conclude that female Sprague-Dawley rats may be marginally stressed when present in the same room in which decapitation is being performed, but similar stress like responses are induced by common husbandry and experimental procedures. Finally, group housing often, but not always, reduces the stress-like responses that can occur in female by-stander rats. PMID- 12580571 TI - Evaluation of postoperative analgesia in a rat model of incisional pain. AB - Effective postoperative analgesia is essential for improving patient well-being and decreasing morbidity. Historical recommendations of postoperative analgesics have been based on their effectiveness in attenuating a nociceptive response in animals that have not undergone a surgical procedure, potentially leading to over or underestimation of postoperative analgesia requirements. This study was designed to evaluate the efficacy of four analgesics in a model of postsurgical pain, which involves surgical incision of the plantar aspect of the hindpaw in halothane-anesthetized rats. The hindpaw was selected as the injury site because it permits quantitative assessment of mechanical sensitivity, which increases as a consequence of tissue damage. As the primary endpoints for postoperative recovery, mechanical sensitivity and weight gain were determined for 5 days. Analgesic regimens included buprenorphine (0.025, 0.05, and 0.1 mg/kg subcutaneously [s.c.]; 1 ml/kg), fentanyl (0.01 and 0.1 mg/kg intraperitoneally [i.p.]; 1 ml/kg), flunixin meglumine (1.1 and 2.5 mg/kg, s.c.; 1 ml/kg) and acetaminophen (100 and 300 mg/kg orally; approximately 3 & 10 ml/kg). Drugs were administered once daily on days 0, 1, and 2 postoperatively. Buprenorphine, fentanyl, and flunixin all significantly decreased mechanical sensitivity, but buprenorphine provided the highest degree of analgesia during the postoperative treatment period. However, rats treated with buprenorphine exhibited heightened mechanical sensitivity once treatment was discontinued on day 2. Moreover, buprenorphine also compromised weight gain as compared to that of vehicle-treated animals. These findings suggest that potent nonsteroidal anti-inflammatory agents, such as flunixin, may be useful alternatives to opioid-based agents for the control of acute postoperative pain associated with a minor surgical procedure and highlight the importance of assessing the risk-benefit ratio when selecting analgesics and dosing regimens. PMID- 12580572 TI - The effects of confinement environment on reproductive efficiency in the baboon. AB - Historically, to supply animals for medical research, both captive-bred baboons and imported wild-caught animals have been available. Now that imported animals are difficult to obtain, it is important to maximize domestic production. To this end, it is necessary to determine the optimum housing conditions (i.e., environmentally dependent factors) under which baboons have the greatest reproductive efficiency. At our institution, the Oklahoma University Health Sciences Center Primate Facility, we recently moved the majority of our baboon breeding colony into a large indoor-outdoor facility (El Reno) from a small, indoor facility (Annex). Fortuitously, this move allowed a direct comparison of baboon reproductive efficiency between the two radically different environments. The environment at the Annex is exclusively indoor and possesses limited but adequate living space (per Guide for the Care and Use of Laboratory Animals recommendations), whereas the El Reno environment is indoor-outdoor and naturalistic with living space exceeding Guide recommendations. Although we expected animals at the El Reno facility to exhibit somewhat increased reproductive efficiency, the magnitude of the increase was surprising: the mean number of days post-partum to first estrus was 165 for animals housed in the Annex, but 69 for those at the El Reno facility. In addition, the mean number of days from first estrus to conception was 61 for baboons in the Annex compared with 47 for those at El Reno, and the mean number of days from conception to next conception was 403 for animals in the Annex but 296 for those at El Reno. These results demonstrate that a change in housing environment can dramatically increase baboon breeding efficiency. PMID- 12580573 TI - Development of a quantitative method for evaluation of the electroencephalogram of rats by using radiotelemetry. AB - The objective of this study was to develop a method for recording the electroencephalogram (EEG) of rats by using radiotelemetry followed by automated analysis of the raw tracing by fast Fourier transformation (FFT). We surgically implanted four 4- to 5-month-old Wistar (Crl:(WI)BR) rats each with two epidural electrodes and an attached radiotransmitter (Physiotel Implant, model TA11CTA, Data Sciences International, Inc., St. Paul, Minn.). Raw EEG signals were routed to a digital storage oscilloscope (DataSys 7200, Gould Instrument Systems, Valley View, Ohio) with FFT and averaging capabilities. FFTs of raw signals were generated at baseline (predose) and after intraperitoneal dosing of the rats with atropine (30 min postdose; 6 mg/kg), caffeine (90 and 150 min postdose; 30 mg/kg), ketamine (15 and 30 min postdose; 50 mg/kg), and pentobarbarbital (60 and 90 min postdose; 40 mg/kg). For atropine, caffeine, and pentobarbital, the overall direction of the change in power across the spectrum (0 to 40 Hz) was the same as that reported previously. Relative peaks in power were consistent with those previous reports for atropine at < or =5 Hz and caffeine at approximately 7.5 Hz. Ketamine caused a shift in peak power from 5 to 10 Hz to < 5 Hz, as shown by other investigators. FFT analysis of EEG signals captured by radiotelemetry may provide a meaningful method for studying the CNS effect of novel compounds. PMID- 12580574 TI - Aspiration method to collect epithelial cells from mouse, rat, and monkey oviducts. AB - When oviduct epithelial cells are collected by using enzymatic methods that involve protease such as trypsin, other cells unintentionally are collected as well, especially from small laboratory animals such as mice and rats, thus contaminating the collected sample. We therefore developed a simple nonenzymatic method that involves using a glass micropipette to aspirate the oviduct epithelial cells from the oviduct. This aspiration method easily removed oviduct epithelial cells from mice, rats, and cynomolgus monkeys. Culture of the collected oviduct epithelial cells confirmed that the cells could adhere and grow on a petri dish. Although this aspiration technique was developed for use in small animals, our results show that it can be applied to cynomolgus monkeys. The advantages of this technique are its simplicity and its success in collecting oviduct epithelial cells from oviducts. PMID- 12580575 TI - Coinfection of laboratory rats with Mycoplasma pulmonis and Chlamydia pneumoniae. AB - Routine examinations of conventional outbred Wistar rats in our laboratory showed increased serum levels of alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and urea. Electron microscopy and specific reactions showed C. pneumoniae and M. pulmonis in lung, liver, spleen, heart, and kidney sections. We could not exclude the fact that other infectious microorganisms detected through routine health surveillance affected the Wistar rat colony; however, we have not identified any of those microorganisms by electron microscopy of the organs listed. Natural coinfection of C. pneumoniae and M. pulmonis can occur in laboratory rats and is associated with histopathological and functional compromise of many organs. Further studies comparing different conventional animals and specific pathogen-free animals are necessary to better understand the present findings and to define whether coinfection influences the results of experimental studies with rats. PMID- 12580576 TI - Open-field assessment of spontaneous activity. PMID- 12580577 TI - Uniaxial alignment of the columnar super-structure of a hexa (alkyl) hexa-peri hexabenzocoronene on untreated glass by simple solution processing. AB - Uniaxially aligned, thin films of a discotic columnar thermotropic liquid crystalline material can be prepared by a simple solution zone-casting method, without the need for modified surfaces or traditional alignment techniques. Atomic force microscopy together with X-ray diffraction reveals single crystalline-like order over several square centimeters, far exceeding the requirements for application of such films in organic molecular electronic devices such as field-effect transistors. PMID- 12580578 TI - Activity of Candida rugosa lipase immobilized on gamma-Fe2O3 magnetic nanoparticles. AB - We report the stability and enzymatic activity of Candida rugosa Lipase (E.C.3.1.1.3) immobilized on gamma-Fe2O3 magnetic nanoparticles. The immobilization strategies were either reacting the enzyme amine group with a nanoparticle surface acetyl, or amine groups. In the former, the enzyme was attached through a C=N bond, while in the latter it was connected using glutaraldehyde. AFM images show an average particle size of 20 +/- 10 nm after deconvolution. The enzymatic activity of the immobilized lipase was determined by following the ester cleavage of p-nitrophenol butyrate. The covalently immobilized enzyme was stabile and reactive over 30 days. PMID- 12580579 TI - Nanoscale soldering of metal nanoparticles for construction of higher-order structures. AB - We demonstrated that platinum nanoparticles dispersed in pure water were "soldered" by gold into higher-order structures such as "nanowebs". In practice, gold nanoparticles in water containing the platinum nanoparticles were melted by irradiation of a pulsed 532-nm laser, which excites selectively the surface plasmon band of the gold nanoparticles, and the melted gold nanoparticles solder the platinum nanoparticles together into the nanowebs. Optical absorption spectroscopy and electron microscopy together with electron probe microanalysis were employed to observe the structure of the nanowebs and to elucidate their formation dynamics. PMID- 12580580 TI - A planar tetracoordinate carbon and unusual bonding in an organodimetallic propynylidene complex arising from double C-H activation of an allene ligand. AB - Reduction of the organoditantalum allene complex (eta-C5Me4R)2Ta2(mu-X)X3(mu eta1,eta3-C3H4) (R = Me (Cp*), Et; X = Cl, Br) with sodium amalgam leads to the propynylidene complex (eta-C5Me4R)2Ta2(mu-H)2X2(mu-HCCCH) by a formal double 1,3 C-H activation of the allene ligand. The solid-state molecular structure contains a planar HCCCH ligand bridging, in parallel coordination mode, the two tantalum atoms, with the HCCCH and Ta atoms coplanar. Key structural features are a Ta-Ta distance of 2.8817(7) A, propynylidene C-C-C angle of 153.7(13) degrees , C-C distance of 1.370(8) A, Ta-C(central) distance of 2.194(9) A, and Ta-C(terminal) distance of 1.970(9) A. Molecular orbital calculations on the complex at the RHF/SBK(d) and B3LYP/LanL2dz levels of theory demonstrate that the propynylidene ligand is best viewed formally as an allenediylidene(4-) ligand bonded to two d0 tantalum atoms via two Ta=C(terminal) double bonds and an unusual three-center, two-electron bridge bond involving both tantalum atoms and a lone pair on the planar, tetracoordinate central carbon. There is no net Ta-Ta bonding based on the orbital analysis. PMID- 12580581 TI - Determination of the enantiomeric composition of guest molecules by chemometric analysis of the UV-visible spectra of cyclodextrin guest-host complexes. AB - Multivariate statistical techniques were applied to the UV spectra of a series of solutions at pH 12 containing a fixed concentration (30 mM) of beta-cyclodextrin (beta-CD) and a fixed concentration (15 mM) of 2-phenylglycine (phi-Gly) with various known enantiomeric compositions. Multivariate correlation of the spectral data for the solutions containing the phi-Gly/beta-CD guest-host complexes with the known enantiomeric composition of the phi-Gly samples was accomplished by partial-least-squares regression. When the multivariate model was used to predict the enantiomeric purity of a test set of samples over the mol fraction range of 0.5-0.9 R-phi-Gly, the average magnitude of the relative errors in the mol fraction determination of enantiomeric composition was 3%. A plot of the enantiomeric composition predicted by the model versus the known enantiomeric composition of the calibration set gave a straight line with a correlation coefficient of 0.955, a slope of 1.05, and an offset of 5.61 x 10-4. PMID- 12580582 TI - Catalytic enantioselective addition of dialkylzinc to N diphenylphosphinoylimines. A practical synthesis of alpha-chiral amines. AB - The enantioselective addition of dialkylzinc reagents to N diphenylphosphinoylimines derived from aryl-, furyl-, and cyclopropylaldehydes is efficiently catalyzed by a copper(II) triflate/(R,R)-MeDUPHOS complex. The yields are high (51-98%), and the enantiomeric excesses vary from 85 to 96%. This route provides a practical route to alpha-chiral amines. PMID- 12580583 TI - Synthesis and characterization of the monomeric imides Ar'MNAr' ' (M = Ga or In; Ar' or Ar' ' = terphenyl ligands) with two-coordinate gallium and indium. AB - Reaction of Ar'MMAr' (M = Ga or In) with N3Ar' ' (Ar' = C6H3-2,6-Dipp2, Dipp = C6H3-2,6-Pri2, Ar' ' = C6H3-2,6(Xyl-4-But)2) afforded the first monomeric imides of heavier group 13 elements with two-coordinate metals. Planar, trans-bent structures with short M-N bond distances were observed, which are consistent with lone pair character at both M and N and a bond order less than the formally expected triple one. PMID- 12580584 TI - Palladium(0)-catalyzed amination, Stille coupling, and Suzuki coupling of electron-deficient aryl fluorides. AB - The amination of 2-fluoronitrobenzene was Pd(0) catalyzed at 65 degrees C in DMF, and the effectiveness of the catalysis was ligand-dependent. Among the five catalyst systems investigated, Pd(PPh3)4 was the most effective catalyst. The control experiments revealed that Pd(OAc)2 or PPh3 was not responsible for the catalysis. 4-Fluoro-3-nitro-benzonitrile and 4-fluoro-3-nitro-benzaldehyde also underwent Stille coupling and Suzuki coupling in the presence of Pd(PPh3)4, and the reactions afforded the coupling products in 28-86% yields. The control experiments showed no sign of reaction in the absence of palladium. These results were in agreement with the oxidative addition/reductive elimination pathway, where the oxidative addition could conceivably proceed via the SNAr mechanism. PMID- 12580585 TI - A ruthenium-catalyzed reaction of aromatic ketones with arylboronates: a new method for the arylation of aromatic compounds via C-H bond cleavage. AB - The ruthenium-catalyzed reaction of aromatic ketones with arylboronic acid esters (arylboronates) gave the ortho arylation product. For this coupling reaction, a RuH2(CO)(PPh3)3 complex exhibited the highest catalytic activity among the complexes screened. Several aromatic ketones, for example, acetophenones, acetonaphthone, alpha-tetralone, and benzosuberone, can be used in this coupling reaction. A variety of arylboronates containing electron-donating (OMe and NMe2) and -withdrawing (F and CF3) groups were found to react with aromatic ketones to give the corresponding aylation products. The corresponding arylboronic acids could be used in this coupling reaction, but the yields were slightly lower, as compared to those of the reaction using the corresponding arylboronates. PMID- 12580586 TI - Facile synthesis of 2,5-diarylthiazoles via palladium-catalyzed tandem C-H substitutions. Design of tunable light emission and liquid crystalline characteristics. AB - A mild and regioselective C-H substitution reaction of thiazole by the catalysis of the palladium/copper system is found to proceed in the presence of tetrabutylammonium fluoride under mild conditions. Applying the new reaction, we synthesized various 2,5-diarylthiazoles that exhibit light-emitting and liquid crystalline characteristics in good yields. The color of light emission is tunable by the introduction of the substituent. Differently substituted 2,5 diarylthiazoles are found to exhibit a stable liquid crystal phase in a wide temperature range. PMID- 12580587 TI - A method for the generation of glycoprotein mimetics. AB - A general method for preparing glycoprotein mimetics with defined glycan structure using the Z domain protein as an example is reported. An unnatural amino acid containing the keto group was site-specifically incorporated into a target protein, Z domain, in response to the amber nonsense codon with high translational fidelity and efficiency. An aminooxy saccharide derivative was then selectively coupled to this genetically encoded keto group. The resulting saccharide core was elaborated to a glycan complex with glycosyltransferases. Alternatively, aminooxy analogues of more complicated glycans were prepared and directly attached to the keto group. Homogeneous glycoprotein mimetics thus prepared should prove useful for the study of carbohydrate effects on glycoprotein structure and function. This method may also lead to the production of glycoprotein therapeutics from Escherichia coli. PMID- 12580588 TI - Tetrathiomolybdate causes formation of hepatic copper-molybdenum clusters in an animal model of Wilson's disease. AB - Wilson's disease is an autosomal recessive human illness in which large quantities of copper accumulate in various organs, including the brain and the liver. If left untreated, it results in hepatitis, neurological complications, and death. Long-Evans Cinnamon (LEC) rats have a homologous mutation to Wilson's disease and thus provide an animal model. Liver lysosomes from tetrathiomolybdate treated LEC rats were isolated and analyzed by Cu and Mo K-edge X-ray absorption spectroscopy. The lysosomes contained a Cu-Mo-S cluster in which the Mo is coordinated by four sulfurs at 2.24 A with approximately three copper neighbors at 2.70 A. Each Cu is coordinated to 3-4 sulfurs at 2.28 A with approximately one Mo neighbor at 2.70 A. These results indicate the formation of a biologically novel molybdenum-copper-sulfur cluster. PMID- 12580589 TI - Isotope effects on the picosecond time-resolved emission spectroscopy of tris(2,2'-bipyridine)ruthenium (II). AB - The excited-state properties of the transition metal complexes tris(2,2' bipyridine) ruthenium(II) and tris(4,7-diphenyl-1,10-phenanthroline) ruthenium(II) are examined using picosecond time-resolved luminescence spectroscopy. For both complexes, direct observation of a short-lived high-energy emission with a lifetime of less than 4 ps is reported. Upon deuteriation of the complexes the lifetime of the high-energy emission shows a marked increase with a biexponential decay (20 and approximately 300 ps components). Examination by time resolved excited-state resonance Raman shows that for the perprotio complexes features attributable to the 3MLCT excited state are formed within 4 ps, while for the perdeuterio a rise time of approximately 20 ps is observed in the 3MLCT features. The results indicate that the emission in both cases may be 1MLCT in origin and are discussed with respect to heterogeneous electron transfer. PMID- 12580590 TI - Bis(((S)-binaphthoxy)(isopropoxy)titanium) oxide as a mu-oxo-type chiral Lewis acid: application to catalytic asymmetric allylation of aldehydes. AB - A new, chiral bis-Ti(IV) oxide of type 1 was successfully designed and can be utilized for strong activation of aldehyde carbonyls, thereby allowing a new catalytic enantioselective allylation of aldehydes with allyltributyltin. The chiral bis-Ti(IV) catalyst (S,S)-1 can be readily prepared either by treatment of bis(triisopropoxy)titanium oxide with (S)-binaphthol or by reaction of ((S) binaphthoxy)isopropoxytitanium chloride with silver(I) oxide. Reaction of hydrocinnamaldehyde with allyltributyltin (1.1 equiv) under the influence of in situ generated chiral bis-Ti(IV) oxide (S,S)-1 (10 mol %) in CH2Cl2 at 0 degrees C for 4 h afforded 1-phenyl-5-hexen-3-ol in 84% yield with 99% ee. The present asymmetric allylation using nonracemic bis-Ti(IV) oxide 1 with partially resolved (S)-binaphthol exhibits a positive nonlinear effect in correlating the enantiopurity of allylation product with the ee of (S)-binaphthol. This asymmetric approach provides a very useful way for obtaining high reactivity and selectivity by the simple introduction of the M-O-M unit in the design of chiral Lewis acid catalysts. PMID- 12580591 TI - Direct observation of insertion events at rac-(C2H4(1-indenyl)2)Zr(MeB(C6F5)3) polymeryl intermediates: distinction between continuous and intermittent propagation modes. AB - The propagating species for 1-hexene, propene, and ethene polymerization as catalyzed by [rac-(C2H4(1-indenyl)2)Zr(CH3)][CH3B(C6F5)3] has been intercepted at T < -40 degrees C and characterized by NMR methods. Observation of the propagating species permits direct monitoring of initiation, propagation, and termination processes by NMR. Detailed examination of alkene incorporation into Zr-polymeryl species using 13C-labeled 1-alkenes supports a continuous insertion mechanism whereby the anion re-coordinates after each alkene insertion. PMID- 12580592 TI - Total synthesis of (+)-phomactin a using a B-alkyl Suzuki macrocyclization. AB - A total synthesis of (+)-phomactin A is described using a B-alkyl Suzuki macrocyclization to incorporate the isolated trisubstituted olefin. This macrocyclization was accomplished with the sensitive hydrated furan ring in place. (R)-(+)-pulegone was used to establish the highly substituted cyclohexene core of the molecule. PMID- 12580593 TI - Theoretical study of halophilic reactions of stable silylenes with chloro- and bromocarbons. AB - A theoretical study of the mechanism of the reaction of stable silylenes with halocarbons has been carried out using the B3LYP density functional method. The main findings are as follows: (1) Lewis acid-base complexes formed between silylenes and halocarbons do not play a role in silylene insertion chemistry into halocarbons; therefore, the acid-base complex mechanism proposed by West et al. (J. Am. Chem. Soc. 2002, 124, 4186) is not appropriate to describe the disilane formation reaction. (2) The disilane formation reactions follow the energetically favorable general reaction pathway (X = halogen): (i) Y2Si: + HCX3 --> TS1 --> Y2XSi-CHX2. (ii) Y2Si: + Y2XSi-CHX2 --> TS2 --> Y2XSi-SiY2CHX2. (3) The observed preference of stable silylenes to undergo C-X bond insertion rather than C-H bond has been investigated. The theoretical findings suggest that this preference is a result of the thermodynamic factor. (4) Stable silylenes prefer to insert into a C-Br rather than a C-Cl bond because the energy barrier to insertion is lower, and the reaction is more exothermic. PMID- 12580594 TI - Microscopic hydration of the sodium chloride ion pair. AB - Hydration of ion pairs is an essential process in various physicochemical phenomena occurring in solutions. Isolated clusters of an ion pair solvated with finite number of waters have been considered as a model system for the critical evaluation of microscopic interactions involved in the process, and theoretical studies have contributed exclusively to the subject up to now. Here we report the first experimental characterization of structure and internal dynamics of hydrated ion pairs, NaCl-(H2O)n (n = 1-3). The measurements of their rotational spectra have proven that the clusters have cyclic forms, in which Na+ and Cl- ions are strongly interacted with the O and H atoms of the solvent molecules, respectively. The Na-Cl distance shows a pronounced increase with the successive addition of water molecules. The separation for n = 3 approaches the value predicted for the contact ion-pair state in aqueous solution by recent molecular dynamics simulations. PMID- 12580595 TI - delta-[Ru(acac)(2)L] (L = a mesogenic derivative of bpy) as a novel chiral dopant for nematic liquid crystals with large helical twisting power. AB - The helical twisting power of a chiral ruthenium complex Delta-[Ru(acac)2L], in which L (mesogenic ligand) and acac denote 5,5'-(4-octylphenyloxycarbonyl)-2,2' bipyridyl and acetylacetonate, respectively, was determined to be betaM = -71 and -1.8 x 102 mum-1 in room-temperature nematics ZLI-1132 and MBBA, respectively. PMID- 12580596 TI - "The great escape" from antiaromaticity: reduction of strained pyrenes. AB - The two-electron reduction of strained pyrenes ([7](2,7)pyrenophanes) with lithium metal leads to the formation of a new sigma-bond as a means to "escape" strained antiaromaticity. PMID- 12580597 TI - Potentials of mean force between ionizable amino acid side chains in water. AB - Potentials of mean force (PMF) between all possible ionizable amino acid side chain pairs in various protonation states were calculated using explicit solvent molecular dynamics simulations with umbrella sampling and the weighted histogram analysis method. The side chains were constrained in various orientations inside a spherical cluster of 200 water molecules. Beglov and Roux's Spherical Solvent Boundary Potential was used to account for the solvent outside this sphere. This approach was first validated by calculating PMFs between monatomic ions (K(+), Na(+), Cl(-)) and comparing them to results from the literature and results obtained using Ewald summation. The strongest interaction (-4.5 kcal/mol) was found for the coaxial Arg(+).Glu(-) pair. Many like-charged side chains display a remarkable lack of repulsion, and occasionally a weak attraction. The PMFs are compared to effective energy curves obtained with common implicit solvation models, namely Generalized Born (GB), EEF1, and uniform dielectric of 80. Overall, the EEF1 curves are too attractive, whereas the GB curves in most cases match the minima of the PMF curves quite well. The uniform dielectric model, despite some fortuitous successes, is grossly inadequate. PMID- 12580598 TI - HADDOCK: a protein-protein docking approach based on biochemical or biophysical information. AB - The structure determination of protein-protein complexes is a rather tedious and lengthy process, by both NMR and X-ray crystallography. Several methods based on docking to study protein complexes have also been well developed over the past few years. Most of these approaches are not driven by experimental data but are based on a combination of energetics and shape complementarity. Here, we present an approach called HADDOCK (High Ambiguity Driven protein-protein Docking) that makes use of biochemical and/or biophysical interaction data such as chemical shift perturbation data resulting from NMR titration experiments or mutagenesis data. This information is introduced as Ambiguous Interaction Restraints (AIRs) to drive the docking process. An AIR is defined as an ambiguous distance between all residues shown to be involved in the interaction. The accuracy of our approach is demonstrated with three molecular complexes. For two of these complexes, for which both the complex and the free protein structures have been solved, NMR titration data were available. Mutagenesis data were used in the last example. In all cases, the best structures generated by HADDOCK, that is, the structures with the lowest intermolecular energies, were the closest to the published structure of the respective complexes (within 2.0 A backbone RMSD). PMID- 12580599 TI - Spectroscopic characterization and assignment of reduction potentials in the tetraheme cytochrome C554 from Nitrosomonas europaea. AB - The tetraheme cytochrome c(554) (cyt c(554)) from Nitrosomonas europaea is an essential electron transfer component in the biological oxidation of ammonia. The protein contains one 5-coordinate heme and three bis-His coordinated hemes in a 3D arrangement common to a newly characterized class of multiheme proteins. The ligand binding, electrochemical properties, and heme-heme interactions are investigated with Mossbauer and X- and Q-band (parallel/perpendicular mode) EPR spectroscopy. The results indicate that the 5-coordinate heme will not bind the common heme ligands, CN(-), F(-), CO, and NO in a wide pH range. Thus, cyt c(554) functions only in electron transfer. Analysis of a series of electrochemically poised and chemically reduced samples allows assignment of reduction potentials for heme 1 through 4 of +47, +47, -147, and -276 mV, respectively. The spectroscopic results indicate that the hemes are weakly exchange-coupled (J approximately -0.5 cm(-)(1)) in two separate pairs and in accordance with the structure: hemes 2/4 (high-spin/low-spin), hemes 1/3 (low-spin/low-spin). There is no evidence of exchange coupling between the pairs. A comparison of the reduction potentials between homologous hemes of cyt c(554) and other members of this new class of multiheme proteins is discussed. Heme 1 has a unique axial N(delta)-His coordination which contributes to a higher potential relative to the homologous hemes of hydroxylamine oxidoreductase (HAO) and the split-Soret cytochrome. Heme 2 is 300 mV more positive than heme 4 of HAO, which is attributed to hydroxide coordination to heme 4 of HAO. PMID- 12580600 TI - Side chain dynamics in unfolded protein states: an NMR based 2H spin relaxation study of delta131delta. AB - NMR relaxation data on disordered proteins can provide insight into both structural and dynamic properties of these molecules. Because of chemical shift degeneracy in correlation spectra, detailed site-specific analyses of side chain dynamics have not been possible. Here, we present new experiments for the measurement of side chain dynamics in methyl-containing residues in unfolded protein states. The pulse schemes are similar to recently proposed methods for measuring deuterium spin relaxation rates in (13)CH(2)D methyl groups in folded proteins.(1) However, because resolution in (1)H-(13)C correlation maps of unfolded proteins is limiting, relaxation data are recorded as a series of (1)H (15)N spectra. The methodology is illustrated with an application to the study of side chain dynamics in delta131delta, a large disordered fragment of staphylococcal nuclease containing residues 1-3 and 13-140 of the wide-type protein. A good correlation between the order parameters of the symmetry axes of the methyl groups and the backbone (1)H-(15)N bond vectors of the same residue is observed. Simulations establish that such a correlation is only possible if the unfolded state is comprised of an ensemble of structures which are not equiprobable. A motional model, which combines wobbling-in-a-cone and Gaussian axial fluctuations, is proposed to estimate chi(1) torsion angle fluctuations, sigma(chi)()1, of Val and Thr residues on the basis of the backbone and side chain order parameters. Values of sigma(chi)()1 are approximately 10 degrees larger than what has previously been observed in folded proteins. Of interest, the value of sigma(chi)()1 for Val 104 is considerably smaller than for other Val or Thr residues, suggesting that it may be part of a hydrophobic cluster. Notably large (15)N transverse relaxation rates are observed in this region. To our knowledge, this is the first time that side chain dynamics in an unfolded state have been studied in detail by NMR. PMID- 12580601 TI - Molecular dynamics simulations and thermodynamics analysis of DNA-drug complexes. Minor groove binding between 4',6-diamidino-2-phenylindole and DNA duplexes in solution. AB - An extended set of nanosecond-scale molecular dynamics simulations of DNA duplex sequences in explicit solvent interacting with the minor groove binding drug 4',6 diamidino-2-phenylindole (DAPI) are investigated for four different and sequence specific binding modes. Force fields for DAPI have been parametrized to properly reflect its internal nonplanarity. Sequences investigated include the binding modes observed experimentally, that is, AATT in d(CGCGAATTCGCG)(2) and ATTG in d(GGCCAATTGG)(2) and alternative shifted binding modes ATTC and AATT, respectively. In each case, stable MD simulations are obtained, well reproducing specific hydration patterns seen in the experiments. In contrast to the 2.4 A d(CGCGAATTCGCG)(2) crystal structure, the DAPI is nonplanar, consistent with its gas-phase geometry and the higher resolution crystal structure. The simulations also suggest that the DAPI molecule is able to adopt different conformational substates accompanied by specific hydration patterns that include long-residing waters. The MM_PBSA technology for estimating relative free energies was utilized. The most consistent free energy results were obtained with an approach that uses a single trajectory of the DNA-DAPI complex to estimate all free energy terms. It is demonstrated that explicit inclusion of a subset of bound water molecules shifts the calculated relative binding free energies in favor of both crystallographically observed binding modes, underlining the importance of structured hydration. PMID- 12580602 TI - Controlled assembly of carbon nanotubes by designed amphiphilic Peptide helices. AB - Carbon nanotubes have properties potentially useful in diverse electrical and mechanical nanoscale devices and for making strong, light materials. However, carbon nanotubes are difficult to solubilize and organize into architectures necessary for many applications. In the present paper, we describe an amphiphilic alpha-helical peptide specifically designed not only to coat and solubilize carbon nanotubes, but also to control the assembly of the peptide-coated nanotubes into macromolecular structures through peptide-peptide interactions between adjacent peptide-wrapped nanotubes. The data presented herein show that the peptide folds into an amphiphilic alpha-helix in the presence of carbon nanotubes and disperses them in aqueous solution by noncovalent interactions with the nanotube surface. Electron microscopy and polarized Raman studies reveal that the peptide-coated nanotubes assemble into fibers with the nanotubes aligned along the fiber axis. Most importantly, the size and morphology of the fibers can be controlled by manipulating solution conditions that affect peptide-peptide interactions. PMID- 12580603 TI - ZP4, an improved neuronal Zn2+ sensor of the Zinpyr family. AB - A second-generation fluorescent sensor for Zn(2+) from the Zinpyr family, ZP4, has been synthesized and characterized. ZP4 (Zinpyr-4, 9-(o-carboxyphenyl)-2 chloro-5-[2-(bis(2-pyridylmethyl)aminomethyl)-N-methylaniline]-6-hydroxy-3 xanthanone) is prepared via a convergent synthetic strategy developed from previous studies with these compounds. ZP4, like its predecessors, has excitation and emission wavelengths in the visible range ( approximately 500 nm), a dissociation constant (K(d)) for Zn(2+) of less than 1 nM and a high quantum yields (Phi = approximately 0.4), making it well suited for biological applications. A 5-fold fluorescent enhancement is observed under simulated physiological conditions corresponding to the binding of the Zn(2+) cation to the sensor, which inhibits a photoinduced electron transfer (PET) quenching pathway. The metal-binding stereochemistry of ZP4 was evaluated through the synthesis and X-ray structural characterization of [M(BPAMP)(H(2)O)(n)](+) complexes, where BPAMP is [2-(bis(2-pyridylmethyl)aminomethyl)-N-methylaniline]-phenol and M = Mn(2+), Zn(2+) (n = 1) or Cu(2+) (n = 0). PMID- 12580604 TI - Preparation and characterization of polypseudorotaxanes based on block-selected inclusion complexation between poly(propylene oxide)-poly(ethylene oxide) poly(propylene oxide) triblock copolymers and alpha-cyclodextrin. AB - A series of new polypseudorotaxanes were synthesized in high yields when the middle poly(ethylene oxide) (PEO) block of poly(propylene oxide)-poly(ethylene oxide)-poly(propylene oxide) (PPO-PEO-PPO) triblock copolymers was selectively recognized and included by alpha-cyclodextrin (alpha-CD) to form crystalline inclusion complexes (ICs), although the middle PEO block was flanked by two thicker PPO blocks, and a PPO chain had been previously thought to be impenetrable to alpha-CD. X-ray diffraction studies demonstrated that the IC domains of the polypseudorotaxanes assumed a channel-type structure similar to the necklace-like ICs formed by alpha-CD and PEO homopolymers. Solid-state CP/MAS (13)C NMR studies showed that the alpha-CD molecules in the polypseudorotaxanes adopted a symmetrical conformation due to the formation of ICs. The compositions and stoichiometry of the polypseudorotaxanes were studied using (1)H NMR, and a 2:1 (ethylene oxide unit to alpha-CD) stoichiometry was found for all polypseudorotaxanes although the PPO-PEO-PPO triblock copolymers had different compositions and block lengths, suggesting that only the PEO block was closely included by alpha-CD molecules, whereas the PPO blocks were uncovered. The hypothesis was further supported by the differential scanning calorimetry (DSC) studies of the polypseudorotaxanes. The glass transitions of the PPO blocks in the polypseudorotaxanes were clearly observed because they were uncovered by alpha-CD and remained amorphous, whereas the glass-transition temperatures increased, because the molecular motion of the PPO blocks was restricted by the hard crystalline phases of the IC domains formed by alpha-CD and the PEO blocks. The thermogravimetric analysis (TGA) revealed that the polypseudorotaxanes had better thermal stability than their free components due to the inclusion complexation. Finally, the kinetics of the threading process of alpha-CD onto the copolymers was also studied. The findings reported in this article suggested interesting possibilities in designing other cyclodextrin ICs and polypseudorotaxanes with block structures. PMID- 12580605 TI - Isolating benzenium ion salts. AB - When partnered with carborane anions, arenium ions are remarkably stable. Previously investigated only at subambient temperatures in highly superacidic media, protonated benzene is readily isolated as a crystalline salt, thermally stable to >150 degrees C. Salts of the type [H(arene)][carborane] have been prepared by protonating benzene, toluene, m-xylene, mesitylene, and hexamethylbenzene with the carborane superacid H(CB(11)HR(5)X(6)) (R = H, Me; X = Cl, Br). They have been characterized by elemental analysis, X-ray crystallography, NMR and IR methods. Solid-state (13)C NMR spectra are similar to those observed earlier in solution, indicating that lattice interactions are comparable to solution solvation effects. The acidic proton(s) of the arenium cations interact weakly with the halide substituents of the anion via ion pairing. This is reflected in the dependence of the C-H stretching frequency on the basicity of the carborane anion. Bond lengths in the arenium ions are consistent with predominant cyclohexadienyl cation character, but charge distribution within the cation is less well represented by this resonance form. Structural and vibrational comparison to theory is made for the benzenium ion (C(6)H(7)(+)) with density functional theory at B3LYP/6-31G and B3P86/6 311+G(d,p) levels. The stability of these salts elevates arenium ions from the status of transients (Wheland intermediates) to reagents. They have been used to bracket the solution-phase basicity of C(60) between that of mesitylene and xylene. PMID- 12580606 TI - Layer-by-layer self-assembled chitosan/poly(thiophene-3-acetic acid) and organophosphorus hydrolase multilayers. AB - The aim of this study is to immobilize an enzyme, namely, organophosphorus hydrolase (OPH), and to detect the presence of paraoxon, which is an organophosphorus compound, using the layer-by-layer (LbL) deposition technique. To lift the OPH from the solid substrate, a pair of polyelectrolytes (positively charged chitosan (CS) and negatively charged poly(thiophene-3-acetic acid) (PTAA)) were combined. These species were made charged by altering the pH of the solutions. LbL involved alternate adsorption of the oppositely charged polyions from dilute aqueous solutions onto a hydrophilic quartz slide. This polyion cushion was held together by the electrostatic attraction between CS and PTAA. The growing process was monitored by fluorescence spectroscopy. OPH was then adsorbed onto the five-bilayer CS/PTAA system. This five-bilayer macromolecular structure compared to the solid substrate rendered stability to the enzyme by giving functional integrity in addition to the ability to react with paraoxon solutions. The ultimate goal is to use such a system to detect the presence of organophosphorus compounds with speed and sensitivity using the absorption and fluorescence detection methodologies. PMID- 12580607 TI - Electronic degeneracies in symmetric (Jahn-Teller) and nonsymmetric aliphatic radical cations: global topology of sigma-bonded molecules. AB - The ground-state potential surfaces of five aliphatic radical cations are investigated using a spin-pairing model. It is shown that the ground-state surface of an n-atomic system supports several stationary points (minima and transition states, including second-order ones). In addition, there are numerous nuclear configurations at which the ground state is electronically degenerate. The electronic degeneracies due to interactions between atoms bound to the same atom are either 2-fold (conical intersections) or 3-fold degenerate but not of a higher dimension. Each 3-fold degeneracy is accompanied by an even number of conical intersections (four or two). A systematic procedure for locating all of these nuclear configurations (that are in fact 3n - 8 or 3n - 11 dimensional hypersurfaces) is described. The model allows for the qualitative determination of the structure and charge distribution of the system at all of the stationary points and electronic degeneracies. Quantum chemical calculations confirm the predictions of the model, which is used to direct and facilitate the calculations. PMID- 12580608 TI - A new Horner-Wadsworth-Emmons type coupling reaction between nonstabilized beta hydroxy phosphonates and aldehydes or ketones. AB - Treatment of nonstabilized beta-hydroxy phosphonic acid mono methyl esters with diisopropyl carbodiimide at ambient temperature leads to clean stereospecific elimination. The phosphonic acid mono alkyl esters are accessible by the selective partial saponification of dimethyl or diethyl alkyl phosphonates with NaOH or MgBr(2). Isolated yields over both hydrolysis and elimination steps average 55-75%. PMID- 12580609 TI - Kemp elimination in membrane mimetic reaction media: probing catalytic properties of catanionic vesicles formed from double-tailed amphiphiles. AB - The rate-determining deprotonation of 5-nitrobenzisoxazole (Kemp elimination) by hydroxide is efficiently catalyzed by vesicles formed from dimethyldioctadecylammonium chloride (C(18)()C(18)()(+)()). Gradual addition of sodium didecyl phosphate (C(10)()C(10)()(-)()) leads to the formation of catanionic vesicles, which were characterized by cryo-electron microscopy, and their main phase transition temperatures (DSC) and zeta-potentials. Increasing percentages of C(10)()C(10)()(-)() in the vesicular bilayers decrease the catalysis of the Kemp elimination. A detailed kinetic analysis, supported by consideration of substrate binding site polarities and counterion binding percentages, suggest that the catalytic effects of C(18)()C(18)()(+)()/C(10)()C(10)()(-)() catanionic vesicles are primarily determined by the binding of catalytically active hydroxide ions to the vesicular surface area. The formation of neutral microdomains between 10 and 30 mol % of C(10)()C(10)()(-)() in the bilayer, as revealed by DSC, is not apparent from the catalytic effects found for these vesicles. Interestingly, the catalytic effects observed for 50 mol % C(10)()C(10)()(-)() in the catanionic vesicles indicate an asymmetric distribution of C(18)()C(18)()(+)() and C(10)()C(10)()(-)() over the bilayer leaflets. The overall kinetic results illustrate the highly complex mix of factors which determines catalytic effects on reactions occurring in biological cell membranes. PMID- 12580610 TI - Group 6 heteroatom- and non-heteroatom-stabilized carbene complexes. beta,beta'- and alpha,beta,beta'-annulation reactions of cyclic enamines. AB - Cyclization reactions of group 6 Fischer carbene complexes with cyclopentanone and cyclohexanone enamines are described. Enamine 3a undergoes thermal alpha,beta,beta'-annulation with alkenylcarbene complexes 1 and 2 (THF, 60 degrees C), affording semibullvalenes 5. The metalate intermediates 6, resulting from beta,beta'-annulation of the enamines 3a and 4a, were quantitatively formed by running the reaction in hexane at room temperature. Acid-promoted demetalation of 6 afforded endo-2-bicyclo[3.2.1]octen-8-ones 7 and endo/exo-2 bicyclo[3.3.1]nonen-9-ones 8 (endo/exo = 5:1). Using (S)-methoxymethylpyrrolidine derived enamines 3b and 4b,c allowed highly enantioenriched cycloadducts endo-(+) 7 as well as endo-(-)-8 and exo-(-)-8 to be accessed. The non-heteroatom stabilized carbene complex 10 was formed from complex 6 by Me(3)SiOTf-promoted elimination of the methoxy group, characterized by (13)C NMR, and transformed into the organic compounds 7, 7-d, and 11 as well as into bicyclo[3.2.1]octan-2,8 diones 14 and cycloheptanones 15. On the basis of this sequence, enantioenriched cycloheptanones (+)-15 were efficiently prepared in one pot from carbene complexes 2 and enamine 3b (51-55% yield, 91-96% ee). Extension of this work to simple Fischer carbene complexes 16 allowed an appropriate way to generate the nonstabilized pentacarbonyl[(phenyl(alkyl)carbene]tungsten complex 17 to be designed, for which the thermal and chemical behavior leading to compounds 18-21 is described. PMID- 12580611 TI - Total synthesis of (+)-4,5-deoxyneodolabelline. AB - The first total synthesis of the marine dolabellane diterpene (+)-4,5 deoxyneodolabelline (1) has been accomplished. The highly efficient approach is characterized by the convergent assembly of dihydropyrans 2ab and cyclopentylsilanes 3ab. Allylic silane 3a was prepared from 2-methyl-2 cyclopentenone via a copper-catalyzed 1,4-addition followed by diastereoselective alkylation of the resulting enolate. A chemical resolution of racemic cyclopentanone 8 was effected by (S)-CBS-catalyzed borohydride reduction. Direct hydroxymethylation of the enantioenriched ketone 8 to form allylic alcohol 14 was achieved by a Stille palladium-catalyzed cross-coupling from the cyclopentenyl triflate 13. Treatment of the corresponding allylic phosphate 15 with trimethylsilylcopper reagent provided for displacement with allylic transposition yielding the exocyclic allylsilane 3a with excellent diastereoselectivity. Dihydropyrans 2a and 2b were prepared from optically pure acyclic acetals via ring-closing metathesis. Coupling of 3a and 2a or 2b via the carbon-Ferrier protocol gave trans-2,6-disubstituted dihydropyrans 30 and 35 with complete stereoselectivity. Vanadium-based pinacol coupling reactions were explored for closure of the medium-sized carbocycle to yield syn-diol 33. X-ray diffraction studies of the monobenzoate 34 have provided unambiguous stereochemical assignments. Substantial ring strain accounted for the lack of alkene products typical of reductive elimination using TiCl(3) and zinc-copper couple (McMurry) conditions leading to 37. Finally, the natural product 1 was obtained via Swern oxidation of the diol 37. PMID- 12580613 TI - Ordered polyelectrolyte multilayers. Rules governing layering in organic binary multilayers. AB - We study the growth and internal structure of polyelectrolyte multilayers obtained by combining three polyanions with nine polycations of the ionene family, of systematically varied chemical architecture. We find that, contrary to a generally held belief, ordered organic multilayers are by no way exceptional, provided one of the polyelectrolytes bears groups which induce structure in water, such as long hydrophobic segments or mesogenic groups. However, this condition is not sufficient, as order will or will not emerge in the multilayer depending on the specific pairing of the polyelectrolytes. The results support the notion that layering in the multilayer results from some degree of prestructuring of a water-swollen layer adsorbed during each step of deposition. These findings pave the way to new possible uses of polyelectrolyte multilayers, for example, for applications requiring preferential alignment or strong confinement of specific functional groups. PMID- 12580612 TI - Proton inventory study of the base-catalyzed hydrolysis of formamide. Consideration of the nucleophilic and general base mechanisms. AB - An NMR study of the rates of hydroxide-promoted hydrolysis of formamide in aqueous media of varying mole fraction D(2)O (n) was performed at [LO(-)] = 1.42 M, T = 25 degrees C, to shed light on whether the mechanism involves a nucleophilic attack of HO(-) on the C=O or HO(-) acting as a general base to remove a proton from an attacking water. The solvent deuterium kinetic isotope effect under these conditions is inverse, k(OH)/k(OD) = 0.77 +/- 0.02 or k(OD)/k(OH) = 1.30 +/- 0.03. Proton inventory analysis of the k(n)() versus n data was undertaken through NLLSQ fits to equations representing four possible mechanisms encompassing nucleophilic and general base ones with waters of solvation on the attacking hydroxide, and with or without waters of solvation on the developing amide hydrate oxyanion. Both nucleophilic and general base mechanisms can be accommodated, but there are restraints on each that are discussed. The preferred mechanism is a nucleophilic one proceeding through a transition state having two solvating waters remaining on the attacking hydroxide and three additional waters attached to the developing amide hydrate oxyanion. PMID- 12580614 TI - Proton-coupled O-O activation on a redox platform bearing a hydrogen-bonding scaffold. AB - Porphyrin architectures bearing a hydrogen-bonding scaffold have been synthesized. The H-bond pendant allows proton-coupled electron transfer (PCET) to be utilized as a vehicle for effecting catalytic O-O bond activation chemistry. Suzuki cross-coupling reactions provide a modular synthetic strategy for the attachment of porphyrins to a rigid xanthene or dibenzofuran pillar bearing the H bond pendant. The resulting HPX (hanging porphyrin xanthene) and HPD (hanging porphyrin dibenzofuran) systems permit both the orientation and acid-base properties of the hanging H-bonding group to be controlled. Comparative reactivity studies for the catalase-like disproportionation of hydrogen peroxide and the epoxidation of olefins by the HPX and HPD platforms with acid and ester hanging groups reveal that the introduction of a proton-transfer network, properly oriented to a redox-active platform, can orchestrate catalytic O-O bond activation. For the catalase and epoxidation reaction types, a marked reactivity enhancement is observed for the xanthene-bridged platform appended with a pendant carboxylic acid group, establishing that this approach can yield superior catalysts to analogues that do not control both proton and electron inventories. PMID- 12580615 TI - Total synthesis of the ramoplanin A2 and ramoplanose aglycon. AB - Full details of a convergent total synthesis of the ramoplanin A2 and ramoplanose aglycon are disclosed. Three key subunits composed of residues 3-9 (heptapeptide 15), pentadepsipeptide 26 (residues 1, 2 and 15-17), and pentapeptide 34 (residues 10-14) were prepared, sequentially coupled, and cyclized to provide the 49-membered depsipeptide core of the aglycon. Key to the preparation of the pentadepsipeptide 26 incorporating the backbone ester was the asymmetric synthesis of an orthogonally protected l-threo-beta-hydroxyasparagine and the development of effective and near-racemization free conditions for esterification of its hindered alcohol (EDCI, DMAP, 0 degrees C). The coupling sites were chosen to maximize the convergency of the synthesis including that of the three subunits, to prevent late stage racemization of carboxylate-activated phenylglycine-derived residues, and to enlist beta-sheet preorganization of an acyclic macrocyclization substrate for 49-membered ring closure. By altering the order of final couplings, two macrocyclization sites, Phe(9)-d-Orn(10) and Gly(14)-Leu(15), were examined. Macrocyclization at the highly successful Phe(9) d-Orn(10) site (89%) may benefit from both beta-sheet preorganization as well as closure at a d-amine terminus within the confines of a beta-turn at the end of the H-bonded antiparallel beta-strands. A more modest, but acceptable macrocyclization reaction at the Gly(14)-Leu(15) site (40-50%) found at the other end of the H-bonded antiparallel beta-strands within a small flexible loop may also benefit from preorganization of the cyclization substrate, is conducted on a substrate incapable of competitive racemization, and accommodates the convergent preparation of analogues bearing depsipeptide modifications. Deliberate late stage incorporation of the subunit bearing the labile depsipeptide ester and a final stage Asn(1) side-chain introduction provides future access to analogues of the aglycons which themselves are equally potent or more potent than the natural products in antimicrobial assays. PMID- 12580616 TI - Conformational effects on optical rotation. 3-substituted 1-butenes. AB - A calculation of the optical rotation of (R)-(-)-3-chloro-1-butene found a remarkably large dependence on the C=C-C-C torsional angle. At tau = 0 degrees, [alpha](D) = +244 degrees, whereas at tau = 180 degrees, [alpha](D) = -526 degrees. The effect of conformation on the optical rotation was confirmed by a study of the temperature dependence of the rotation. An analysis of the data gave the difference in free energy between the low- and high-energy conformers as 1315 cal/mol and gave the optical rotation of the low-energy conformer and the average of the rotations of the higher energy forms. Although a large effect was found, the observed rotations are a factor of 2.6 smaller than the calculated values, independent of both conformation and wavelength from 589 to 365 nm. The effect of replacing Cl with F, CN, and CCH was examined theoretically. The effects of substituents are remarkably small despite large changes in the calculated electronic transition energies. PMID- 12580617 TI - Synthesis, structure, and magnetic properties of the fullerene-based ferromagnets Eu3C70 and Eu9C70. AB - Intercalation of C(70) with europium affords two kinds of magnetic compounds, a canted antiferromagnet Eu(x)C(70) (x approximately 3) and a ferromagnet Eu(x)C(70) (x approximately 9) with transition temperatures (T(C)) of 5 and 38 K, respectively. The Curie constants in the paramagnetic phase and the saturation moment in the ferromagnetic phase are both understood by the full moment of Eu(2+) for both systems. The structure of Eu(3)(-)(delta)C(70) (delta approximately 0.27) is pseudo-monoclinic, derived by a simple deformation of the parent face-centered cubic (fcc) structure. Eu(9)(-)(delta)C(70) (delta approximately 0.2) forms an fcc structure, in which cuboctahedral clustering of Eu(2+) ions is observed in the enhanced size octahedral holes. The observed T(C) of the Eu(9)(-)(delta)C(70) ferromagnet is comparable to or larger than those of simple binary Eu-based ferromagnets, such as Eu chalcogenides or carbides, despite the low atomic ratio of Eu in the chemical formulas. This can be understood by the short Eu(2+)-Eu(2+) distances and high coordination numbers permitted by the multiple occupation by Eu(2+) ions of the expanded octahedral interstitial sites in higher fullerene-based solids. PMID- 12580618 TI - Identification of active sites in gold-catalyzed hydrogenation of acrolein. AB - The active sites of supported gold catalysts, favoring the adsorption of C=O groups of acrolein and subsequent reaction to allyl alcohol, have been identified as edges of gold nanoparticles. After our recent finding that this reaction preferentially occurs on single crystalline particles rather than multiply twinned ones, this paper reports on a new approach to distinguish different features of the gold particle morphology. Elucidation of the active site issue cannot be simply done by varying the size of gold particles, since the effects of faceting and multiply twinned particles may interfere. Therefore, modification of the gold particle surface by indium has been used to vary the active site characteristics of a suitable catalyst, and a selective decoration of gold particle faces has been observed, leaving edges free. This is in contradiction to theoretical predictions, suggesting a preferred occupation of the low-coordinated edges of the gold particles. On the bimetallic catalyst, the desired allyl alcohol is the main product (selectivity 63%; temperature 593 K, total pressure p(total) = 2 MPa). From the experimentally proven correlation between surface structure and catalytic behavior, the edges of single crystalline gold particles have been identified as active sites for the preferred C=O hydrogenation. PMID- 12580619 TI - MIL-50, an open-framework GaPO with a periodic pattern of small water ponds and dry rubidium atoms: a combined XRD, NMR, and computational study. AB - A new fluorinated gallium phosphate, MIL-50, has been synthesized under mild hydrothermal conditions using 1,6-diaminohexane. The chemical formula of MIL-50 is Rb(2)Ga(9)(PO(4))(8)(HPO(4))(OH)F(6).2N(2)C(6)H(18).7H(2)O. The structure is a network of hexameric units of Ga(3)(PO(4))(3)F(2) and Ga(3)(PO(4))(2)(HPO(4))F(3) via corner sharing. It creates a three-dimensional open-framework delimiting 6- and 18-ring channels running along the c axis. The diprotonated 1,6-diaminohexane and water molecules are trapped within the 18-ring pores, whereas the rubidium cations reside in the 6-ring ones. A double quantum (31)P NMR experiment and partial charge calculations indicate that water molecules are present under the form of periodic small clusters, lowering the multiplicity of one phosphorus site, P3. Though water hops within the clusters, the motion leaves the water pattern periodic. Rubidium is so tightly embedded into the framework that water moving in the large 18-ring channels does not reach it, leaving it therefore dry. The crystal framework may be ascribed to the orthorhombic space group Cmc2(1) (n degrees 36), a = 32.1510(2), b = 17.2290(3), c = 10.2120(1) A. The periodic water pattern has a different symmetry than that of the framework. A method has been devised to superpose the two sublattices that coexist in the same unit cell in order to have full occupancy of each site and to perform Madelung summations. This original method is of general interest for most zeolitic materials exhibiting a different symmetry for the framework and the template sublattices. PMID- 12580620 TI - Density functional and reduction potential calculations of Fe4S4 clusters. AB - Density functional theory geometry optimizations and reduction potential calculations are reported for all five known oxidation states of [Fe(4)S(4)(SCH(3))(4)](n)()(-) (n = 0, 1, 2, 3, 4) clusters that form the active sites of iron-sulfur proteins. The geometry-optimized structures tend to be slightly expanded relative to experiment, with the best comparison found in the [Fe(4)S(4)(SCH(3))(4)](2)(-) model cluster, having bond lengths 0.03 A longer on average than experimentally observed. Environmental effects are modeled with a continuum dielectric, allowing the solvent contribution to the reduction potential to be calculated. The calculated protein plus solvent effects on the reduction potentials of seven proteins (including high potential iron proteins, ferredoxins, the iron protein of nitrogenase, and the "X", "A", and "B" centers of photosystem I) are also examined. A good correlation between predicted and measured absolute reduction potentials for each oxidation state of the cluster is found, both for relative potentials within a given oxidation state and for the absolute potentials for all known couples. These calculations suggest that the number of amide dipole and hydrogen bonding interactions with the Fe(4)S(4) clusters play a key role in modulating the accessible redox couple. For the [Fe(4)S(4)](0) (all-ferrous) system, the experimentally observed S = 4 state is calculated to lie lowest in energy, and the predicted geometry and electronic properties for this state correlate well with the EXAFS and Mossbauer data. Cluster geometries are also predicted for the [Fe(4)S(4)](4+) (all-ferric) system, and the calculated reduction potential for the [Fe(4)S(4)(SCH(3))(4)](1)( )(/0) redox couple is in good agreement with that estimated for experimental model clusters containing alkylthiolate ligands. PMID- 12580621 TI - Investigation of Zr-C, Zr-N, and potential agostic interactions in an organozirconium complex by experimental electron density analysis. AB - The crystal structure and electron density (ED) distribution of an imine coupling product with an open zirconocene, Zr(2,4-C(7)H(11))[(i Pr)NCHPhCH(2)CMe=CHCMe=CH(2)] (C(7)H(11) = dimethylpentadienyl), have been derived from accurate synchrotron X-ray diffraction measurements. The molecular structure reveals asymmetric coordination of Zr by the pentadienyl (2,4 C(7)H(11)) ligand ( = 2.56(6) A), the butadiene fragment ( = 2.43(5) A), and the amide nitrogen atom (Zr-N = 2.0312(5) A) of the second ligand. The study of the ED and its topological analysis affords new insight into the bonding and electronic structure of the title zirconium complex. The interactions between the metal center and the ligands are represented by a Zr-N bond path and one Zr-C bond path with each of the pentadienyl and butadiene moieties, contrary to the usually depicted global metal-ligand bonding. The butadiene and pentadienyl groups exhibit a polarization of the corresponding pi-like ED in the C-Zr directions, indicating that the whole conjugated systems are nonetheless involved. The 4d atomic orbitals of Zr exhibit unusual populations according to ligand field considerations, which reveal a high degree of sigma-donation from the conjugated pi systems of both ligands. As deduced from numerical integration over the topologically defined atomic basins, the Zr to ligand charge transfer is 1.48 e to the C(17)NH(24) ligand and 0.68 e to dimethylpentadienyl. Topological analysis of a short intramolecular Zr.(C,H) contact provides no indication of the presence of agostic interactions, despite a small Zr-N-C angle of 102.87(4) degrees. Thus, no bond path and BCP (bond critical point) of the ED are found in the Zr.(C,H) region, which would have provided evidence for such direct interactions, nor is there any evidence for charge accumulation between the Zr and H atoms, or for lengthening of the C-H bond involved in the putative interaction. These characteristics, similar to those in other distorted situations, may be common for other electron-deficient d(0) complexes. PMID- 12580622 TI - Molecular dynamics simulations of end-to-end contact formation in hydrocarbon chains in water and aqueous urea solution. AB - We probe the urea-denaturation mechanism using molecular dynamics simulations of an elementary "folding" event, namely, the formation of end-to-end contact in the linear hydrocarbon chain (HC) CH(3)(CH(2))(18)CH(3). Electrostatic effects are examined using a model HC in which one end of the chain is positively charged (+0.2e) and the other contains a negative charge (-0.2e). For these systems multiple transitions between "folded" (conformations in which the chain ends are in contact) and "unfolded" (end-to-end contact is broken) can be observed during 4 ns molecular dynamics simulations. In water and 6 M aqueous urea solution HC and the charged HC fluctuate between collapsed globular conformations and a set of expanded structures. The collapsed conformation adopted by the HC in water is slightly destablized in 6 M urea. In contrast, the end-to-end contact is disrupted in the charged HC only in aqueous urea solution. Despite the presence of a large hydrophobic patch, on length scales on the order of approximately 8-10 A "denaturation" (transition to the expanded unfolded state) occurs by a direct interaction of urea with charges on the chain ends. The contiguous patch of hydrophobic moieties leads to "mild dewetting", which becomes more pronounced in the charged HC in 6 M aqueous urea solution. Our simulations establish that the urea denaturation mechanism is most likely electrostatic in origin. PMID- 12580623 TI - General rules for predicting where a catalytic reaction should occur on metal surfaces: a density functional theory study of C-H and C-O bond breaking/making on flat, stepped, and kinked metal surfaces. AB - To predict where a catalytic reaction should occur is a fundamental issue scientifically. Technologically, it is also important because it can facilitate the catalyst's design. However, to date, the understanding of this issue is rather limited. In this work, two types of reactions, CH(4) <--> CH(3) + H and CO <--> C + O on two transition metal surfaces, were chosen as model systems aiming to address in general where a catalytic reaction should occur. The dissociations of CH(4) --> CH(3) + H and CO --> C + O and their reverse reactions on flat, stepped, and kinked Rh and Pd surfaces were studied in detail. We find the following: First, for the CH(4) <--> Ch(3) + H reaction, the dissociation barrier is reduced by approximately 0.3 eV on steps and kinks as compared to that on flat surfaces. On the other hand, there is essentially no difference in barrier for the association reaction of CH(3) + H on the flat surfaces and the defects. Second, for the CO <--> C + O reaction, the dissociation barrier decreases dramatically (more than 0.8 eV on Rh and Pd) on steps and kinks as compared to that on flat surfaces. In contrast to the CH(3) + H reaction, the C + O association reaction also preferentially occurs on steps and kinks. We also present a detailed analysis of the reaction barriers in which each barrier is decomposed quantitatively into a local electronic effect and a geometrical effect. Our DFT calculations show that surface defects such as steps and kinks can largely facilitate bond breaking, while whether the surface defects could promote bond formation depends on the individual reaction as well as the particular metal. The physical origin of these trends is identified and discussed. On the basis of our results, we arrive at some simple rules with respect to where a reaction should occur: (i) defects such as steps are always favored for dissociation reactions as compared to flat surfaces; and (ii) the reaction site of the association reactions is largely related to the magnitude of the bonding competition effect, which is determined by the reactant and metal valency. Reactions with high valency reactants are more likely to occur on defects (more structure-sensitive), as compared to reactions with low valency reactants. Moreover, the reactions on late transition metals are more likely to proceed on defects than those on the early transition metals. PMID- 12580624 TI - Ab initio molecular orbital study on the G-selectivity of GGG triplet in copper(I)-mediated one-electron oxidation. AB - The G-selectivity for Cu(I)-mediated one-electron oxidation of 5'-TG(1)G(2)G(3) 3' and 5'-CG(1)G(2)G(3)-3' has been examined by ab initio molecular orbital calculations. It was confirmed that G(1) is selectively damaged by Cu(I) ion for both 5'-TG(1)G(2)G(3)-3' and 5'-CG(1)G(2)G(3)-3', being good agreement with experimental results. The Cu(I)-mediated G(1)-selectivity is primarily due to the stability of the Cu(I)-coordinated complex, [-XG(1)G(2)G(3)-, Cu(I)(H(2)O)(3)](+). The Cu(I) ion coordinates selectively to N7 of G(2) of 5' G(1)G(2)G(3)-3' rather than N7 of G(1). The G(2)-selective coordination induces the G(1)-selective trap of a hole that is created by one-electron oxidation and migrates to GGG triplet. Therefore, the radical cation of G(1) is selectively created in both 5'-TG(1)G(2)G(3)-3' and 5'-CG(1)G(2)G(3)-3', giving the G(1) selective damage of 5'-G(1)G(2)G(3)-3'. PMID- 12580625 TI - Enantiomeric excesses and electronic chirality measure. AB - We present an approach to measure the amount of chirality in the electronic wave function and apply it to the study of a set of asymmetric aminohydroxylation reactions. The correlation coefficient between the chirality measure and the enantiomeric excesses is larger than 0.9 and suggests that this phenomenological approach can be a valuable tool to investigate and, perhaps in the future, design asymmetric synthesis. PMID- 12580626 TI - Probing the dynamic guest-host interactions in sol-gel films using single molecule spectroscopy. AB - Organic dyes usually exhibit enhanced photostability when trapped inside sol-gel silicates. The enhanced photostability is attributed to the reduction of intramolecular motions that facilitate photodegradation. We report the simultaneous detection of mobility and photostability of sol-gel encapsulated didodecyl-3,3,3',3'-tetramethylindocarbocyanine (DiI) using single molecule spectroscopy. Fluorescence from DiI was resolved into parallel and perpendicular polarization components and separately detected. On the basis of the calculated fluorescence polarization, single DiI molecules were classified into "tumbling" and "fixed". Out of 212 molecules investigated, 52% were found to be fixed. For the first time, the mobility of a guest molecule in sol-gel silicate can be directly correlated with its own photostability. Both tumbling and fixed molecules have shown to exhibit nonuniform photostability, indicative of the very heterogeneous guest-host interactions within each subgroup. The survival lifetimes for the majority of the tumbling and fixed molecules were found to be 4.3 and 13.1 s, respectively, demonstrating unequivocally that fixed molecules exhibit a higher photostability than tumbling molecules. These results are in accordance with a recent study on rhodamine B encapsulated in dried sol-gel silicates. PMID- 12580627 TI - Physical and kinetic analysis of the cooperative role of metal ions in catalysis of phosphodiester cleavage by a dinuclear Zn(II) complex. AB - A dinuclear metal ion complex Zn(2)()(L2O) and its mononuclear analogue Zn(L1OH) were synthesized and studied as catalysts of the cleavage of the phosphate diester 2-hydroxypropyl-4-nitrophenyl phosphate (HPNP). X-ray crystal structure data, potentiometric titrations, and (1)H NMR spectra obtained over a wide range of pH values provide strong evidence that the alcohol linker in the complex Zn(2)()(L2O) is ionized below pH 6.0, while the alcohol group in the complex Zn(L1OH) remains protonated even at high pH. The ionizations observed at high pH correspond to the formation of the monohydroxo complexes, Zn(2)(L2O)(OH) and Zn(L1OH)(OH), with pK(a)'s of 8.0 and 9.2, respectively. The pH-rate profiles of second-order rate constants for metal-ion complex-catalyzed cleavage of HPNP are reported. These show downward curvature centered at the pK(a)'s for the respective zinc-bound waters, and limiting second-order rate constants at high pH of k(c) = 0.71 M(-)(1) s(-)(1) for Zn(2)()(L2O) and 0.061 M(-)(1) s(-)(1) for Zn(L1OH). The larger catalytic activity of Zn(2)()(L2O) compared with Zn(L1OH) is due to the cooperative role of the metal ions in facilitating the formation of the ionized zinc-bound water at close to neutral pH and in providing additional stabilization of the rate-limiting transition state for phosphodiester cleavage. Zn(2)()(L2O) complex (1 M) at pH 7.6 stabilizes the transition state for the uncatalyzed reaction by 9.3 kcal/mol. Assuming that the dissociation constant determined for a diethyl phosphate inhibitor is similar to that for substrate, then ca. 2.4 kcal/mol of these stabilizing interactions are expressed in the ground-state Michaelis complex, while the bulk of these interactions are only expressed as the reaction approaches the transition state for phosphodiester cleavage. PMID- 12580628 TI - Experimental study of the heterogeneous interaction of SO3 and H2O: formation of condensed phase molecular sulfuric acid hydrates. AB - The interaction of SO(3) and H(2)O at low temperatures upon an inert surface has been studied with infrared spectroscopy and compared to the predictions of recent computational studies. At low temperatures and low water partial pressures, amorphous deposits of molecular H(2)SO(4) complexed with variable amounts of H(2)O in a ratio of between 1:1 and 2:1 are formed. Upon annealing, this material ejects water and converts first to a 1:1 H(2)SO(4).H(2)O complex and subsequently to anhydrous H(2)SO(4). Adding water to the amorphous molecular hydrate results in the formation of a new species, which on the basis of its thermal behavior and by comparison to theoretical predictions can be attributed to a molecular polymer with a repeat unit of (H(2)SO(4).(H(2)O)(2))(n)(). Implications of these observations for the initial stages of the formation of sulfate aerosol in the atmosphere and their surface reactivity are discussed. PMID- 12580629 TI - Heterogeneous electron-transfer kinetics for ruthenium and ferrocene redox moieties through alkanethiol monolayers on gold. AB - The standard heterogeneous electron-transfer rate constants between substrate gold electrodes and either ferrocene or pentaaminepyridine ruthenium redox couples attached to the electrode surface by various lengths of an alkanethiol bridge as a constituent of a mixed self-assembled monolayer were measured as a function of temperature. The ferrocene was either directly attached to the alkanethiol bridge or attached through an ester (CO(2)) linkage. For long bridge lengths (containing more than 11 methylene groups) the rate constants were measured using either chronoamperometry or cyclic voltammetry; for the shorter bridges, the indirect laser induced temperature jump technique was employed to measure the rate constants. Analysis of the distance (bridge length) dependence of the preexponential factors obtained from an Arrhenius analysis of the rate constant versus temperature data demonstrates a clear limiting behavior at a surprisingly small value of this preexponential factor (much lower than would be expected on the basis of aqueous solvent dynamics). This limit is independent of both the identity of the redox couple and the nature of the linkage of the couple to the bridge, and it is definitely different (smaller) from the limit derived from an equivalent analysis of the rate constant (versus temperature) data for the interfacial electron-transfer reaction through oligophenylenevinylene bridges between gold electrodes and ferrocene. There are a number of possible explanations for this behavior including, for example, the possible effects of bridge conformational flexibility upon the electron-transfer kinetics. Nevertheless, conventional ideas regarding electronic coupling through alkane bridges and solvent dynamics are insufficient to explain the results reported here. PMID- 12580635 TI - Laser-induced material ejection from model molecular solids and liquids: mechanisms, implications, and applications. PMID- 12580634 TI - Computer simulations of laser ablation of molecular substrates. PMID- 12580636 TI - The desorption process in MALDI. PMID- 12580637 TI - Ion formation in MALDI: the cluster ionization mechanism. PMID- 12580638 TI - MALDI ionization: the role of in-plume processes. PMID- 12580640 TI - Photomechanical processes and effects in ablation. PMID- 12580639 TI - Chemical and spectroscopic aspects of polymer ablation: special features and novel directions. PMID- 12580641 TI - Models for laser ablation of polymers. PMID- 12580642 TI - Laser deposition of polymer and biomaterial films. PMID- 12580643 TI - Mechanisms of pulsed laser ablation of biological tissues. PMID- 12580644 TI - The science and politics of medicines control. AB - Drug development and regulation are often presented as purely matters of technical science. In this paper it is argued that, in principle, toxicology, clinical pharmacology and pharmacovigilance in drug testing and regulation are necessarily a combination of science and politics. This has important implications for how one attempts to make progress in drug regulation, such as in interpreting technical evidence and in the setting of regulatory standards with which evidence should be evaluated. In practice, drug testing and regulation are shown to be hybrids of science and politics. Moreover, drawing on existing empirical evidence, it is suggested that this mixture currently, and for some time, has had the wrong ingredients for optimal drug safety and public health outcomes. For example, too often the balance of the scientific doubts about drug safety are weighed to the interests of manufacturers rather than to those of patients and public health, while some scientific standards with which drug safety is to be interpreted are being reshaped in ways that give insufficient priority to the protection of public health. Finally, it is proposed that: drug regulation should include comparative efficacy testing; regulatory agencies should conduct some key tests, charging the costs to industry and without duplication; and the regulatory system should be less secretive and more accountable to public scrutiny. Greater efforts should be made to eliminate experts' conflicts of interest within the regulatory process. PMID- 12580645 TI - Methods and pitfalls in searching drug safety databases utilising the Medical Dictionary for Regulatory Activities (MedDRA). AB - The Medical Dictionary for Regulatory Activities (MedDRA) is a unified standard terminology for recording and reporting adverse drug event data. Its introduction is widely seen as a significant improvement on the previous situation, where a multitude of terminologies of widely varying scope and quality were in use. However, there are some complexities that may cause difficulties, and these will form the focus for this paper. Two methods of searching MedDRA-coded databases are described: searching based on term selection from all of MedDRA and searching based on terms in the safety database. There are several potential traps for the unwary in safety searches. There may be multiple locations of relevant terms within a system organ class (SOC) and lack of recognition of appropriate group terms; the user may think that group terms are more inclusive than is the case. MedDRA may distribute terms relevant to one medical condition across several primary SOCs. If the database supports the MedDRA model, it is possible to perform multiaxial searching: while this may help find terms that might have been missed, it is still necessary to consider the entire contents of the SOCs to find all relevant terms and there are many instances of incomplete secondary linkages. It is important to adjust for multiaxiality if data are presented using primary and secondary locations. Other sources for errors in searching are non-intuitive placement and the selection of terms as preferred terms (PTs) that may not be widely recognised. Some MedDRA rules could also result in errors in data retrieval if the individual is unaware of these: in particular, the lack of multiaxial linkages for the Investigations SOC, Social circumstances SOC and Surgical and medical procedures SOC and the requirement that a PT may only be present under one High Level Term (HLT) and one High Level Group Term (HLGT) within any single SOC. Special Search Categories (collections of PTs assembled from various SOCs by searching all of MedDRA) are limited by the small number available and by lack of clarity about criteria applied in their construction. Difficulties in database searching may be addressed by suitable user training and experience, and by central reporting of detected deficiencies in MedDRA. Other remedies may include regulatory guidance on implementation and use of MedDRA. Further systematic review of MedDRA is needed and generation of standardised searches that may be used 'off the shelf' will help, particularly where the same search is performed repeatedly on multiple data sets. Until these enhancements are widely available, MedDRA users should take great care when searching a safety database to ensure that cases are not inadvertently missed. PMID- 12580646 TI - Quantitative methods in pharmacovigilance: focus on signal detection. AB - Pharmacovigilance serves to detect previously unrecognised adverse events associated with the use of medicines. The simplest method for detecting signals of such events is crude inspection of lists of spontaneously reported drug-event combinations. Quantitative and automated numerator-based methods such as Bayesian data mining can supplement or supplant these methods. The theoretical basis and limitations of these methods should be understood by drug safety professionals, and automated methods should not be automatically accepted. Published evaluations of these techniques are mainly limited to large regulatory databases, and performance characteristics may differ in smaller safety databases of drug developers. Head-to-head comparisons of the major techniques have not been published. Regardless of previous statistical training, pharmacovigilance practitioners should understand how these methods work. The mathematical basis of these techniques should not obscure the numerous confounders and biases inherent in the data. This article seeks to make automated signal detection methods transparent to drug safety professionals of various backgrounds. This is accomplished by first providing a brief overview of the evolution of signal detection followed by a series of sections devoted to the methods with the greatest utilisation and evidentiary support: proportional reporting rations, the Bayesian Confidence Propagation Neural Network and empirical Bayes screening. Sophisticated yet intuitive explanations are provided for each method, supported by figures in which the underlying statistical concepts are explored. Finally the strengths, limitations, pitfalls and outstanding unresolved issues are discussed. Pharmacovigilance specialists should not be intimidated by the mathematics. Understanding the theoretical basis of these methods should enhance the effective assessment and possible implementation of these techniques by drug safety professionals. PMID- 12580647 TI - Safety of celecoxib in individuals allergic to sulfonamide: a pilot study. AB - OBJECTIVE: To evaluate cross reactivity between sulfonamide antimicrobials and celecoxib in patients with histories of allergies to sulfonamide antimicrobials. METHODS: Immunocompetent patients with a history of sulfonamide antimicrobial allergy who were being considered for therapy with celecoxib were prospectively enrolled. Sulfamethoxazole and trimethoprim skin prick and intradermal testing and/or an in vitro lymphocyte toxicity assay were performed. If skin testing was negative, an oral challenge with sulfamethoxazole and trimethoprim was performed. Oral challenges with celecoxib were administered to all patients. RESULTS: Twenty eight immunocompetent patients (26 female; mean age 60 years) were evaluated. History of sulfonamide antimicrobial allergy included urticaria (n = 7), cutaneous eruptions (n = 9), and other (n = 12). Four of the 28 patients who were skin prick tested were positive to sulfamethoxazole and two of the ten patients who underwent in vitro testing were positive to sulfamethoxazole. All 28 patients were administered celecoxib and tolerated the medication. Phone call follow up in 25 patients disclosed that 15 patients continued to take celecoxib, while five patients did not take celecoxib following the oral challenge, and five discontinued celecoxib due to adverse effects, lack of drug efficacy or physician preference. CONCLUSIONS: Confusion exists regarding the potential for cross reactivity between sulfonamide antimicrobials and other sulfonamide-containing compounds. The six sulfonamide-allergic patients tolerated celecoxib uneventfully. This pilot study supports the hypothesis that the potential for cross-reactivity between celecoxib and sulfonamide antimicrobials appears to be low. However, further investigations are required to confirm this. PMID- 12580649 TI - The National Football League combine: a reliable predictor of draft status? AB - The performance of 326 collegiate football players attending the 2000 National Football League combine was studied to determine whether draft status could be predicted from performance measurements. The combine measured height and weight along with 9 performance tests: 225-lb bench press test, 10-yd dash, 20-yd dash, 40-yd dash, 20-yd proagility shuttle, 60-yd shuttle, 3-cone drill, broad jump, and vertical jump. Prediction equations were generated for 7 position categories with varying degrees of accuracy-running backs (RBs), r(2) = 1.00; wide receivers (WRs), r(2) = 1.00; offensive linemen, r(2) = 0.70; defensive linemen, r(2) = 0.59; defensive backs (DBs), r(2) = 1.00; linebackers, r(2) = 0.22; and quarterbacks, r(2) = 0.84. The successes of the prediction equations are related to the ability of the individual tests to assess the necessary skills for each position. This study concludes that the combine can be used to accurately predict draft status of RBs, WRs, and DBs. The equations can also be used as a good to fair estimate for other positions. PMID- 12580648 TI - Utilisation and safety of low molecular weight heparins: prospective observational study in medical inpatients. AB - AIMS: Low molecular weight heparins (LMWHs) are widely used as curative or preventive treatments of thromboembolic diseases. The aim of our study was to: investigate the pattern of prescription of LMWHs in different departments of French teaching hospitals; andestimate the incidence of adverse drug reactions (ADRs) induced by LMWHs and associated risk factors for the occurrence of bleeding events. METHODS: This prospective study was performed in two teaching hospitals in Toulouse (south-western France) in March 1999 in different medical wards. All patients receiving a prescription for a LMWH were included in the survey. All data were prospectively recorded in each ward. RESULTS: A total of 334 patients were included. Sex ratio (male/female) was 1.25 and mean age was 72.5 +/- 16.3 years (extremes:18-101). 450 prescriptions for LMWHs were collected (1.34 prescription per patient) and involved mainly enoxaparin (61%), which was more frequently used than tinzaparin in patients over 75 years old (71.7 vs 28.3%; p < 0.0001). Ninety-nine patients received a LMWH for curative treatment (corresponding to 127 prescriptions of which 99 were for enoxaparin and 28 were for tinzaparin [p < 0.0001]). Indications included therapy for deep venous thrombosis, pulmonary embolism, acute coronary syndrome, unstable angina pectoris, non-Q-wave myocardial infarction. Serious renal insufficiency was significantly more frequent in patients from the geriatrics department (p < 0.00001). Enoxaparin was prescribed more frequently in patients with serious or moderate renal insufficiency than tinzaparin (72 vs 61%, p < 0.05). The incidence of LMWHs-induced ADRs was 10.5% occurring in 22 cases during preventive treatment of deep venous thrombosis and in 13 cases during curative therapy. ADRs were classified as 'serious' in 11 cases (31.4%). Reported ADRs were bleeding events (n = 15), thrombocytosis (n = 13), thrombopenia (n = 4) and hepatic cytolysis (n = 1). The mean delay for the occurrence of bleeding effects was 8.0 +/- 9.1 days (range 1-40). Multivariate analysis of the influence of several criteria on the occurrence of haemorrhagic effects showed that the decrease of creatinine clearance (10 ml/min) was associated with an increased haemorrhagic risk (relative risk [RR] = 1.34, 95% CI 1.12-1.65; p < 0.05). Moreover, the risk of adverse bleeding effects increased for patients with a creatinine clearance <20 ml/min (RR = 2.8; 95% CI 1.00-7.8). CONCLUSION: Our data firstly show a different pattern of LMWHs prescription in different clinical wards. Secondly, the risk of bleeding ADRs in patients treated by LMWHs increases significantly with renal function impairment for the two LMWH preparations studied. More pharmacoepidemiological studies are necessary in patients with several risk factors, particularly in elderly people who often have renal impairment, in order to determine the optimal pattern use of each LMWH. PMID- 12580650 TI - Effects of recovery beverages on glycogen restoration and endurance exercise performance. AB - The restorative capacities of a high carbohydrate-protein (CHO-PRO) beverage containing electrolytes and a traditional 6% carbohydrate-electrolyte sports beverage (SB) were assessed after glycogen-depleting exercise. Postexercise ingestion of the CHO-PRO beverage, in comparison with the SB, resulted in a 55% greater time to exhaustion during a subsequent exercise bout at 85% maximum oxygen consumption (VO(2)max). The greater recovery after the intake of the CHO PRO beverage could be because of a greater rate of muscle glycogen storage. Therefore, a second study was designed to investigate the effects of after exercise CHO-PRO and SB supplements on muscle glycogen restoration. Eight endurance-trained cyclists (VO(2)max = 62.1 +/- 2.2 ml.kg(-1) body wt.min(-1)) performed 2 trials consisting of a 2-hour glycogen-depletion ride at 65-75% VO(2)max. Carbohydrate-protein (355 ml; approximately 0.8 g carbohydrate (CHO).kg(-1) body wt and approximately 0.2 g protein.kg(-1) body wt) or SB (355 ml; approximately 0.3 g CHO.kg(-1) body wt) was provided immediately and 2 hours after exercise. Trials were randomized and separated by 7-15 days. Ingestion of the CHO-PRO beverage resulted in a 17% greater plasma glucose response, a 92% greater insulin response, and a 128% greater storage of muscle glycogen (159 +/- 18 and 69 +/- 32 micromol.g(-1) dry weight for CHO-PRO and SB, respectively) compared with the SB (p < 0.05). These findings indicate that the rate of recovery is coupled with the rate of muscle glycogen replenishment and suggest that recovery supplements should be consumed to optimize muscle glycogen synthesis as well as fluid replacement. PMID- 12580651 TI - The effect of liquid carbohydrate ingestion on repeated maximal effort exercise in competitive cyclists. AB - We investigated the effects of carbohydrate ingestion during recovery from high intensity exercise on subsequent high-intensity exercise in trained cyclists. Aerobic power was determined, and the competitive cyclists (N = 7) were familiarized with the 100-kJ test protocol (100 KJ-TEST). The subjects performed a first 100 KJ-TEST (RIDE-1), ingested 0.7 g.(kg body mass)(-1) of Gatorlode (CHO) or placebo (PLC), rested for 60 minutes, and then performed a second 100 KJ TEST (RIDE-2). Blood samples taken before (PRE-1) and after (POST-1) RIDE-1 and before (PRE-2) and after (POST-2) RIDE-2 were analyzed for plasma glucose ([glucose]), lactate, and nonesterified fatty acids ([NEFA]). No significant differences (p > 0.05) were observed between treatments in time to complete RIDE 1 (CHO = 270.3 +/- 29.0 seconds; PLC = 269.9 +/- 33.0 seconds) and RIDE-2 (CHO = 271.7 +/- 26.6 seconds; PLC = 275.3 +/- 30.6 seconds). Plasma [glucose] significantly decreased during the 60-minute recovery for PLC. There was an interaction effect for [NEFA] during recovery, with [NEFA] increasing for PLC and decreasing for CHO. Carbohydrate ingestion after maximal exercise does not appear to influence subsequent short-duration maximal effort exercise in competitive cyclists but does alter plasma [glucose] and [NEFA] relative to a PLC condition. PMID- 12580652 TI - Creatine supplementation and its effect on musculotendinous stiffness and performance. AB - Anecdotal reports suggesting that creatine (Cr) supplementation may cause side effects, such as an increased incidence of muscle strains or tears, require scientific examination. In this study, it was hypothesized that the rapid fluid retention and "dry matter growth" evident after Cr supplementation may cause an increase in musculotendinous stiffness. Intuitively, an increase in musculotendinous stiffness would increase the chance of injury during exercise. Twenty men were randomly allocated to a control or an experimental group and were examined for musculotendinous stiffness of the triceps surae and for numerous performance indices before and after Cr ingestion. The Cr group achieved a significant increase in body mass (79.7 +/- 10.8 kg vs. 80.9 +/- 10.7 kg), counter movement jump height (40.2 +/- 4.8 cm vs. 42.7 +/- 5.9 cm), and 20-cm drop jump height (32.3 +/- 3.3 cm vs. 35.1 +/- 4.8 cm) after supplementation. No increase was found for musculotendinous stiffness at any assessment load. There were no significant changes in any variables within the control group. These findings have both performance- and injury-related implications. Primarily, anecdotal evidence suggesting that Cr supplementation causes muscular strain injuries is not supported by this study. In addition, the increase in jump performance is indicative of performance enhancement in activities requiring maximal power output. PMID- 12580653 TI - The effect of beta-hydroxy beta-methylbutyrate on muscular strength and body composition in collegiate football players. AB - This study assesses the effects of daily beta-hydroxy beta-methylbutyrate (HMB) supplementation on muscular strength (bench press, squats, and power cleans) and body composition (body weight and body fat) among collegiate football players undergoing a strenuous exercise program. Subjects were collegiate football players (n = 35) training under the supervision of certified strength coaches averaging 20 hours of weekly exercise. In the first supplementation period, 16 of the 35 subjects were supplemented with 3 g of HMB per day for 4 weeks; the other 19 received a placebo followed by a 1-week washout period and then a second supplementation period in a randomized double-blind crossover, placebo design. There were no significant changes (p > 0.05) in muscular strength, including bench press, squats, and power cleans, among the subjects. There were also no significant changes (p > 0.05) in body composition, including body fat and body weight. Very little clinical evidence exists for supplementing HMB in athletic populations. PMID- 12580654 TI - Growth and anabolic hormones, leptin, and neuromuscular performance in moderately trained prepubescent athletes and untrained boys. AB - We investigated hormonal regulators of growth and development, leptin levels, body composition, neuromuscular performance, and the associations among them in trained prepubertal athletes (experimental group [EG]) and an untrained control group (CG). Informed consent was obtained from the children and their parents. Their maturation stage was evaluated according to Tanner's criteria. There were no differences between EG and CG in physical characteristics, body mass index (BMI), lean body mass, testosterone (T), sex hormone-binding globulin, free androgen index, growth hormone (GH), hand grip strength, and jumping performance. Leptin levels and percent fat of the EG were significantly lower than those of the CG (p < 0.05-0.005). Leptin levels were significantly correlated to body fat and BMI for both the EG and the CG (r = 0.51-0.79). There is little evidence that leptin has a positive effect on growth and anabolic factors. Sex hormone-binding globulin and GH may explain the variation of leptin in athletes with low T (R(2) = 0.43) and in CG (R(2) = 0.35), respectively. Leptin seems to be a permissive factor for the onset of puberty, and the training background needs an optimal biological maturation to produce significant differences in muscle and power performance. PMID- 12580655 TI - Effects of ribose supplementation on repeated sprint performance in men. AB - This study used a randomized, placebo-controlled, crossover design to evaluate the effects of oral ribose supplementation on short-term anaerobic performance. After familiarization, subjects performed 2 bouts of repeated cycle sprint exercise (six 10-second sprints with 60-second rest periods between sprints) in a single day. After the second exercise, bout subjects ingested 32 g of ribose or cellulose (4 x 8-g doses) during the next 36 hours. After supplementation, subjects returned to the laboratory to perform a single bout of cycle sprinting (as described above). After a 5-day washout period, subjects repeated the protocol, receiving the opposite supplement treatment. Ribose supplementation lead to statistically significant increases in mean power and peak power in sprint 2 (10.9 and 6.6%, respectively) and higher (although not significant) absolute values in sprints 1, 3, and 4. In conclusion, ribose supplementation did not show reproducible increases in performance across all 6 sprints. Therefore, within the framework of this investigation, it appears that ribose supplementation does not have a consistent or substantial effect on anaerobic cycle sprinting. PMID- 12580656 TI - The effects of ibuprofen on delayed muscle soreness and muscular performance after eccentric exercise. AB - The purpose of this study was to examine the effects of ibuprofen on delayed onset muscle soreness (DOMS), indirect markers of muscle damage and muscular performance. Nineteen subjects (their mean [+/- SD] age, height, and weight was 24.6 +/- 3.9 years, 176.2 +/- 11.1 cm, 77.3 +/- 18.7 kg) performed the eccentric leg curl exercise to induce muscle soreness in the hamstrings. Nine subjects took an ibuprofen pill of 400 mg every 8 hours within a period of 48 hours, whereas 10 subjects received a placebo randomly (double blind). White blood cells (WBCs) and creatine kinase (CK) were measured at pre-exercise, 4-6, 24, and 48 hours after exercise and maximal strength (1 repetition maximum). Vertical jump performance and knee flexion range of motion (ROM) were measured at pre-exercise, 24 and 48 hours after exercise. Muscle soreness increased (p < 0.05) in both groups after 24 and 48 hours, although the ibuprofen group yielded a significantly lower value (p < 0.05) after 24 hours. The WBC levels were significantly (p < 0.05) increased 4-6 hours postexercise in both groups with no significant difference (p > 0.05) between the 2 groups. The CK values increased (p < 0.05) in the placebo group at 24 and 48 hours postexercise, whereas no significant differences (p > 0.05) were observed in the ibuprofen group. The CK values of the ibuprofen group were lower (p < 0.05) after 48 hours compared with the placebo group. Maximal strength, vertical jump performance, and knee ROM decreased significantly (p < 0.05) after exercise and at 24 and 48 hours postexercise in both the placebo and the ibuprofen groups with no differences being observed (p > 0.05) between the 2 groups. The results of this study reveal that intake of ibuprofen can decrease muscle soreness induced after eccentric exercise but cannot assist in restoring muscle function. PMID- 12580657 TI - Improvement in running economy after 6 weeks of plyometric training. AB - This study determined whether a 6-week regimen of plyometric training would improve running economy (i.e., the oxygen cost of submaximal running). Eighteen regular but not highly trained distance runners (age = 29 +/- 7 [mean +/- SD] years) were randomly assigned to experimental and control groups. All subjects continued regular running training for 6 weeks; experimental subjects also did plyometric training. Dependent variables measured before and after the 6-week period were economy of running on a level treadmill at 3 velocities (women: 2.23, 2.68, and 3.13 m.s(-1); men: 2.68, 3.13, and 3.58 m.s(-1)),VO(2)max, and indirect indicators of ability of muscles of lower limbs to store and return elastic energy. The last were measurements during jumping tests on an inclined (20 degrees ) sled: maximal jump height with and without countermovement and efficiencies of series of 40 submaximal countermovement and static jumps. The plyometric training improved economy (p < 0.05). Averaged values (m.ml(-1).kg( 1)) for the 3 running speeds were: (a). experimental subjects-5.14 +/- 0.39 pretraining, 5.26 +/- 0.39 posttraining; and (b). control subjects-5.10 +/- 0.36 pretraining, 5.06 +/- 0.36 posttraining. The VO(2)max did not change with training. Plyometric training did not result in changes in jump height or efficiency variables that would have indicated improved ability to store and return elastic energy. We conclude that 6 weeks of plyometric training improves running economy in regular but not highly trained distance runners; the mechanism must still be determined. PMID- 12580658 TI - Acute effects of plyometric exercise on maximum squat performance in male athletes. AB - This study examines the acute effects of plyometric exercise on 1 repetition maximum (RM) squat performance in trained male athletes. Twelve men (mean age +/- SD: 20.5 +/- 1.4 years) volunteered to participate in 3 testing sessions separated by at least 6 days of rest. During each testing session the 1RM was assessed on back squat exercise. Before all 3 trials subjects warmed up on a stationary cycle for 5 minutes and performed static stretching. Subjects then performed 5 submaximal sets of 1-8 repetitions before attempting a 1RM lift. Subjects rested for at least 4 minutes between 1RM trials. During the first testing session (T1) subjects performed a series of sets with increasing load until their 1RM was determined. During the second and third testing sessions subjects performed in counterbalanced order either 3 double-leg tuck jumps (TJ) or 2 depth jumps (DJ) 30 seconds before each 1RM attempt. The average 1RM lifts after T1 and testing sessions with TJ or DJ were 139.6 +/- 29.3 kg, 140.5 +/- 25.6 kg, and 144.5 +/- 30.2 kg, respectively (T1 < DJ; p < 0.05). These data suggest that DJ performed before 1RM testing may enhance squat performance in trained male athletes. PMID- 12580659 TI - Effect of elastic-cord towing on the kinematics of the acceleration phase of sprinting. AB - We studied the specificity of elastic-cord towing by measuring selected kinematics of the acceleration phase of sprinting. Nine collegiate sprinters ran two 20-m maximal sprints (MSs) and towed sprints (TSs) that were recorded on high speed video (180 Hz). Sagittal plane kinematics of a 4-segment model of the right side of the body were digitized for a complete stride at the 15-m point for the fastest trial. Significant (p < 0.001) differences were observed for horizontal velocity of the center of mass (CoM), stride length (SL), and horizontal distance from the CoM of the foot to the CoM of the body. There was no significant difference in stride rate between the MS and TS conditions. Omega-squared analysis showed that elastic-cord towing accounted for most of the variance in acute changes in horizontal velocity (73%), SL (68%), and horizontal position of the CoM at foot contact (64%). Elastic-cord tow training resulted in significant acute changes in sprint kinematics in the acceleration phase of an MS that do not appear to be sprint specific. More research is needed on the specificity of TS training and long-term effects on sprinting performance. PMID- 12580660 TI - Comparison of metabolic and heart rate responses to super slow vs. traditional resistance training. AB - In order to compare the cardiovascular and energy expenditure demands of "Super Slow" (SST) and traditional (TT) resistance training 7 resistance-trained young men (24.3 +/- 3.8 years) had energy expenditure (using indirect calorimetry) and heart rate evaluated during and for 15 minutes after a workout on separate days. Blood lactate levels were also evaluated before and after each intervention. Resting energy expenditure was evaluated in a fasted state using a ventilated canopy prior to any exercise stimulus and 21 to 22 hours after the SST and TT. VO(2) and average heart rate were both significantly higher during the TT than during the SST. The net VO(2) was also significantly higher during the 15 minutes recovery; however, average heart rate was not significantly different between the 2 groups. Total net energy expenditure from oxidative processes was 45% higher for the TT intervention (TT = 155 +/- 28 kcal, and SST = 107 +/- 20 kcal). The significant postexercise lactate difference was almost 2 times greater following the TT than after the SST (TT = 7.9 +/- 1.7 mmol.L(-1).min(-1), and SST = 4.0 +/- 2.0 mmol.L(-1).min(-1)). Finally, adding the estimated energy expenditure of the blood lactate to the net energy expenditure from the VO(2) produced a significant difference that is over 48% greater for the TT intervention (TT = 172 +/- 29 kcal.min(-1), and SST = 116 +/- 22 kcal.min(-1)). No significant repeated measures analysis main effect was found for either resting energy expenditure or respiratory exchange ratio. The metabolic and cardiovascular stimuli were low with SST. Traditional resistance training increases energy expenditure more than SST does and thus may be more beneficial for body weight control. PMID- 12580661 TI - A comparison of linear and daily undulating periodized programs with equated volume and intensity for local muscular endurance. AB - The purpose of this study was to compare linear periodization (LP), daily undulating periodization (DUP), and reverse linear periodization (RLP) for gains in local muscular endurance and strength. Sixty subjects (30 men, 30 women) were randomly assigned to LP, DUP, or RLP groups. Maximal repetitions at 50% of the subject's body weight were recorded for leg extensions as a pretest, midtest, and posttest. Training involved 3 sets (leg extensions) 2 days per week. The LP group performed sets of 25 repetition maximum (RM), 20RM, and 15RM changing every 5 weeks. The RLP group progressed in reverse order (15RM, 20RM, 25RM), changing every 5 weeks. The DUP group adjusted training variables between each workout (25RM, 20RM, 15RM repeated for the 15 weeks). Volume and intensity were equated for each training program. No significant differences were measured in endurance gains between groups (RLP = 73%, LP = 56%, DUP = 55%; p = 0.58). But effect sizes (ES) demonstrated that the RLP treatment (ES = 0.27) was more effective than the LP treatment (control) and the DUP treatment (ES = -0.02) at increasing muscular endurance. Therefore, it was concluded that making gradual increases in volume and gradual decreases in intensity was the most effective program for increasing muscular endurance. PMID- 12580662 TI - The effect of short-term isokinetic training on force and rate of velocity development. AB - This study determines the effects of short-term isokinetic training on rate of velocity development (RVD) and force. Three groups were pre- and posttested for knee extension RVD and force at 1.04 (slow) and 4.18 rad.s(-1) (fast) on a Kin Com dynamometer. The slow and fast groups completed 2 days of velocity-specific training, whereas the control group did not train. Four-way analysis of variance results demonstrated significant (p < 0.05) decreases in RVD between pre- and posttests for the slow group at the slow velocity (RVD-1.25 +/- 0.04 degrees vs. 1.08 +/- 0.03 degrees ) and for the fast group at the fast velocity (RVD-14.24 +/ 0.33 degrees vs. 13.59 +/- 0.29 degrees ). Force exhibited no significant differences between testing days for any group. These results demonstrate that short-term isokinetic training results in velocity-specific RVD improvements. These acute RVD improvements may serve to offset strength deficits in power environments on the basis of the mutable relationship between force and velocity. PMID- 12580663 TI - Effects of different set configurations on barbell velocity and displacement during a clean pull. AB - The effects of 3 types of set configurations (cluster, traditional, and undulating) on barbell kinematics were investigated in the present study. Thirteen men (track and field = 8; Olympic weightlifters = 5) (mean +/- SEM age, 23.4 +/- 1.1 years; height, 181.3 +/- 2.1 cm; body mass, 89.8 +/- 4.2 kg) performed 1 set of 5 repetitions in a cluster, traditional, and undulating fashion at 90 and 120% of their 1 repetition maximum (1RM) power clean (119.0 +/- 4.3 kg). All data were collected at 50 Hz and analyzed with a V-Scope Weightlifting Analysis System. Peak velocity (PV) and peak displacement (PD) were analyzed for each repetition and averaged for each set type. Results indicated that a significantly (p < 0.016) higher PV occurred during the cluster set when compared with the traditional sets at both intensities. PD was significantly higher than traditional sets at the 120% intensity. The present study suggests set configuration can affect PV and PD during clean pulls. PMID- 12580664 TI - Cardiovascular response to punching tempo. AB - Eighteen trained volunteers (12 men and 6 women: age = 22.0 +/- 2.8 years, height = 170.79 +/- 7.67 cm, weight = 71.54 +/- 12.63 kg) participated in 2-minute, randomized fitness boxing trials, wearing 0.34-kg punching gloves, at various tempos (60, 72, 84, 96, 108, and 120 b.min(-1)). During each trial, oxygen uptake (VO(2)), heart rate (HR), and ventilation (VE) were measured continuously. A rating of perceived exertion (RPE) was attained at the conclusion of each trial. Subjects were able to attain VO(2) values ranging from 26.83 to 29.75 ml.kg( 1).min(-1), which correspond to 67.7-72.5% of VO(2)max. The HR responses yielded results ranging from 167.4 to 182.2 b.min(-1), or 85 to 93% of HRmax. No significant difference (p > 0.05) was seen with VO(2) between trials, although a significant difference (p < 0.05) was observed with HR, VE, and RPE. It appears that boxing speed is associated with increased VE, HR response, and perceived effort but not with VO(2). Energy expenditure values ranged from 9.8 to 11.2 kcal.min(-1) for the boxing trials. These results suggest that fitness boxing programs compare favorably with other exercise modalities in cardiovascular response and caloric expenditure. PMID- 12580665 TI - Strength changes during an in-season resistance-training program for football. AB - The purpose of this study was to examine the effects of both intensity and volume of training during a 2 d.wk(-1) in-season resistance-training program (RTP) for American football players. Fifty-three National Collegiate Athletic Association Division III football players were tested in the 1 repetition maximum (1RM) bench press and 1RM squat on the first day of summer training camp (PRE) and during the final week of the regular season (POST). Subjects were required to perform 3 sets of 6-8 repetitions per exercise. Significant strength improvements in squat were observed from PRE (155.0 +/- 31.8 kg) to POST (163.3 +/- 30.0 kg), whereas no PRE to POST changes in bench press were seen (124.7 +/- 21.0 kg vs.123.9 +/- 18.6 kg, respectively). Training volume and training compliance were not related to strength improvement. Further analysis showed that athletes training at >or=80% of their PRE 1RM had significantly greater strength improvements than athletes training at <80% of their PRE 1RM, for both bench press and squat. Strength improvements can be seen in American football players, during an in-season RTP, as long as exercise intensity is >or=80% of the 1RM. PMID- 12580666 TI - The influence of volume of exercise on early adaptations to strength training. AB - The purpose of this investigation was to compare the effects of single-set strength training and 3-set strength training during the early phase of adaptation in 18 untrained male subjects (age, 20-30 years). After initial testing, subjects were randomly assigned to either the 3L-1U group (n = 8), which trained 3 sets in leg exercises and 1 set in upper-body exercises, or the 1L-3U group (n = 10), which trained 1 set in leg exercises and 3 sets in upper-body exercises. Testing was conducted at the beginning and at the end of the study and consisted of 2 maximal isometric tests (knee extension and bench press) and 6 maximal dynamic tests (1 repetition maximum [1RM] tests). Subjects trained 3 days per week for 6 weeks. After warm-up, subjects performed 3 leg exercises and 4 upper-body exercises. In both groups, each set consisted of 7 repetitions (reps) with the load supposed to induce muscular failure after the seventh rep (7RM load). After 6 weeks of training, 1RM performance in all training exercises was significantly increased (10-26%, p < 0.01) in both groups. The relative increase in 1RM load in the 3 leg exercises was significantly greater in the 3L-1U group than in the 1L-3U group (21% vs. 14%, p = 0.01). However, the relative increase in 1RM load in the 3 upper-body exercises was similar in the 3L-1U group (16%) and the 1L-3U group (14%). These results show a superior adaptation to 3-set strength training, compared with 1-set strength training, in leg exercises but not in upper-body exercises during the early phase of adaptation. PMID- 12580667 TI - The initial effects of low-volume strength training on balance in untrained older men and women. AB - Evidence indicates that leg weakness in older adults is associated with decreased control of balance. The gender-specific implications of strength training on control of balance in older men and women remains unknown. This study examined the initial adaptations to 12 weeks of low-volume, single-set-to-failure strength training and its effect on quadriceps strength and control of multidirectional balance in previously untrained older men (n = 11) and women (n = 11) 59-83 years of age. Leg strength increased 23-30% (p < 0.001) across genders; however, the effect on balance varied between genders. No significant changes were noted in the women, whereas 37% (p < 0.014) more sway in the medial-lateral direction was noted in the men, with no change in the anterior-posterior direction. These results demonstrate that this training protocol may not be effective for improving balance and may lead to worsening of balance in older men. PMID- 12580668 TI - Effects of strength training on submaximal and maximal endurance performance capacity in middle-aged and older men. AB - Effects of a 16-week progressive strength-training program on blood lactate accumulation (LA), maximal workload (W(max)) attained during progressive cycling exercise, maximum half-squat (1RM(HS)), muscle cross-sectional area of the quadriceps femoris muscle group (CSA(QF)), and serum hormone concentrations were examined in 11 middle-aged (46 year old [M46]) and 11 older (64 year old [M64]) men. During the 16 weeks of training, significant increases were observed in 1RM(HS) in M46 and M64 (41-45%; p < 0.001). The muscle CSA(QF) increased (13-11%; p > 0.01) for both groups. The first 8 weeks of training led to significant increases in W(max) (6-11%; p < 0.001) and decreases in submaximal (LA) in both groups, but no further training-induced changes were observed during the subsequent 8 weeks of training. Statistically significant relationships were observed in M64 and in the combined group M46 + M64 between the training-induced changes observed in W(max) and serum testosterone-cortisol and free-testosterone cortisol ratios, whereas in M46 the respective correlations values did not reach statistically significant levels. These data indicate that strength training results in a significant improvement in maximal and submaximal endurance during the first 8 weeks of strength training in both age groups, related in part to the intensity and the volume of resistance training used and to the training status of the subjects. The relationships found in this study between various indices of cycling testing and serum hormone concentrations after strength training suggest that maximal incremental cycling might be used as an additional test to detect anabolic-catabolic responses to prolonged strength training in middle-aged and older men. PMID- 12580669 TI - Power and maximum strength relationships during performance of dynamic and static weighted jumps. AB - The purpose of this study was to investigate the relationship of the 1 repetition maximum (1RM) squat to power output during countermovement and static weighted vertical squat jumps. The training experience of subjects (N = 22, 87.0 +/- 15.3 kg, 14.1 +/- 7.1% fat, 22.2 +/- 3.8 years) ranged from 7 weeks to 15+ years. Based on the 1RM squat, subjects were further divided into the 5 strongest and 5 weakest subjects (p 0.70) but not for SQ (r < 0.70). No bodily structural dimension variable had a significant correlation with 1RM strength (r < 0.70). The results of this study indicate that 1RM SQ, BP, and DL may be predicted with an acceptable degree of accuracy in elite male high-school power lifter subjects. PMID- 12580674 TI - Physiological profiles and performance predictors of a women's NCAA rowing team. AB - We described the physiological profiles of rowers (N = 16; age = 20.1 +/- 1.4 years, weight = 78.6 +/- 9.5 kg, height = 177.5 +/- 3.1 cm) of the top 2 varsity boats on an NCAA women's crew and determined whether physiological measures predict boat assignment as determined by the head coach. Eight participants were members of the top varsity boat (1V) and 8 competed at a lower level (2V). Expired gases were collected while subjects completed the U.S. National Team VO(2)max (3-minute stages) and 2 kilometer (2K) time trial rowing ergometer protocols. Heart rates (HR) and blood lactates were measured before, during, and after each test. The VO(2)max and blood lactate at stage 2 of the VO(2)max test were used to predict boat assignment. Average (+/-SD) VO(2)max was 3.86 +/- 0.40 L.min(-1). The 2K times averaged 453.0 +/- 10.5 seconds. Subjects used approximately 96% of VO(2)max and 98% of HR(max) during the 2K time trials. Neither VO(2)max nor submaximal lactate were related to boat assignment. The VO(2) values during the 2K trial indicated that rowing economy differed among athletes. Results of physiological measures should help the coach individualize workouts of top performers. PMID- 12580675 TI - Psychophysics in functional strength and power training: review and implementation framework. AB - Functional training is considered to be training that attempts to mimic the specific physiological demands of real-life activities. Most approaches to functional training, though, omit important factors that contribute to physiological and neuromotor adaptations. Cognitive factors related to sports influence physiological performance, and subsequently, physiological and neuromotor adaptations. We present a rationale and a theoretical framework by which to create effective functional training methods that incorporate cognitive factors. This framework draws upon recent developments and strong empirical evidence in the areas of dynamic systems theory, perceptual skills training, and motor learning/control. Emphasized within rigorous physical training are practice related techniques and motor-learning strategies. In particular, mental effort, attention, and intention manipulated in a discovery-learning paradigm provide a framework for functional strength and power training. This framework is suggested to help maximize sport-specific physiological adaptations, and subsequently, sports performance. PMID- 12580676 TI - Carbohydrate supplementation and resistance training. AB - There is a growing body of evidence suggesting that the performance of resistance training exercises can elicit a significant glycogenolytic effect that potentially could result in performance decrements. These decrements may result in less than optimal physiological adaptations to training. Currently some scientific evidence suggests that carbohydrate supplementation prior to and during high-volume resistance training results in the maintenance of muscle glycogen concentration, which potentially could result in the maintenance or increase of performance during a training bout. Some researchers suggest that ingesting carbohydrate supplements prior to and during resistance training may improve resistance-training performance. Additionally, the ingestion of carbohydrates following resistance exercise enhances the resynthesis of muscle glycogen, which may result in a faster time of recovery from resistance training, thus possibly allowing for a greater training volume. On the basis of the current scientific literature, it may be advisable for athletes who are performing high volume resistance training to ingest carbohydrate supplements before, during, and immediately after resistance training. PMID- 12580677 TI - Treatment and prevention of delayed onset muscle soreness. AB - Eccentric exercise continues to receive attention as a productive means of exercise. Coupled with this has been the heightened study of the damage that occurs in early stages of exposure to eccentric exercise. This is commonly referred to as delayed onset muscle soreness (DOMS). To date, a sound and consistent treatment for DOMS has not been established. Although multiple practices exist for the treatment of DOMS, few have scientific support. Suggested treatments for DOMS are numerous and include pharmaceuticals, herbal remedies, stretching, massage, nutritional supplements, and many more. DOMS is particularly prevalent in resistance training; hence, this article may be of particular interest to the coach, trainer, or physical therapist to aid in selection of efficient treatments. First, we briefly review eccentric exercise and its characteristics and then proceed to a scientific and systematic overview and evaluation of treatments for DOMS. We have classified treatments into 3 sections, namely, pharmacological, conventional rehabilitation approaches, and a third section that collectively evaluates multiple additional practiced treatments. Literature that addresses most directly the question regarding the effectiveness of a particular treatment has been selected. The reader will note that selected treatments such as anti-inflammatory drugs and antioxidants appear to have a potential in the treatment of DOMS. Other conventional approaches, such as massage, ultrasound, and stretching appear less promising. PMID- 12580678 TI - School-based health centers: a long road to travel. PMID- 12580679 TI - Use of psychotropic medications in young, preschool children: primum non nocere. PMID- 12580680 TI - Burden of asthma in inner-city elementary schoolchildren: do school-based health centers make a difference? AB - BACKGROUND: About 1400 school-based health centers (SBHCs) provide care to 1.1 million children. However, it is unknown if access to on-site services is associated with a better outcome. OBJECTIVE: To compare outcomes including hospitalizations, emergency department visits, and school absenteeism in elementary schoolchildren with asthma who were grouped according to their enrollment at schools that have or do not have SBHCs. SETTING: Six elementary schools in The Bronx, NY (4 schools with and 2 without SBHCs). PARTICIPANTS: Nine hundred forty-nine inner-city schoolchildren with asthma. DESIGN: To collect baseline data for a longitudinal study, we surveyed parents to identify children with asthma, and to obtain information about symptoms and the use of health services in the last 12 months. Participating schools provided absenteeism information. RESULTS: Of 6433 parents surveyed, 74% completed the questionnaires. The prevalence of asthma was 19.9% and the morbidity was high-during the previous year, 46.2% had been treated for asthma in an emergency department; 12.6% had been hospitalized. Emergency department use was not associated with SBHCs. However, in univariate and multivariate analyses, the rate of hospitalization was higher among children enrolled at schools without an SBHC (rate ratio, 1.5; 95% confidence interval, 1.1-1.9). In addition, schoolchildren with asthma enrolled in the schools without an SBHC missed more days of school than those enrolled in schools with an SBHC (mean [SD], 21.3 [15.4] vs 18.2 [13.0], respectively; P =.02). CONCLUSION: Access to SBHCs was associated with a reduction in the rate of hospitalization and a gain of 3 days of school for schoolchildren who have asthma. PMID- 12580681 TI - Drug-exposed infants and developmental outcome: effects of a home intervention and ongoing maternal drug use. AB - OBJECTIVE: To evaluate the effects of a home intervention and ongoing maternal drug use on the developmental outcome of drug-exposed infants. DESIGN: Longitudinal randomized cohort study of a home intervention with substance abusing mothers and their infants. Mother-infant dyads were randomly assigned to a control or intervention group at 2 weeks' post partum. Control families received brief monthly tracking visits. Intervention families received weekly home visits from 0 to 6 months and biweekly visits from 6 to 18 months by trained lay visitors. PARTICIPANTS: One hundred eight low-income, inner-city, drug exposed children (control, 54; intervention, 54) who underwent developmental testing at 6, 12, and 18 months post partum and who remained with their biological mothers through 18 months. MAIN OUTCOME MEASURES: Infant scores from the Bayley Scales of Infant Development (BSID) at 6, 12, and 18 months post partum. Maternal report of drug use during the pregnancy and ongoing drug use through 18 months post partum was assessed. RESULTS: In the repeated-measures analyses, intervention infants had significantly higher BSID Mental Developmental Index (MDI) and Psychomotor Developmental Index scores than control infants. Ongoing maternal cocaine and/or heroin use was associated with lower MDI scores. Finally, MDI scores decreased significantly in both groups. CONCLUSIONS: Ongoing maternal drug use is associated with worse developmental outcomes among a group of drug-exposed infants. A home intervention led to higher BSID scores among drug exposed infants. However, BSID MDI scores decreased during the first 18 months post partum among inner-city, low-socioeconomic-status infants in the present study. PMID- 12580682 TI - Quantitative ultrasound of the tibia and radius in prepubertal and early-pubertal female athletes. AB - BACKGROUND: Physical exercise during childhood has been shown to enhance bone mineral density, thus reducing the risk of osteoporosis. OBJECTIVE: To examine bone properties, as measured by quantitative ultrasound, in prepubertal and early pubertal female athletes engaged in impact and nonimpact sports. DESIGN: Survey. SETTING: General community. PARTICIPANTS: Twenty-five acrobatic gymnasts, 21 swimmers, and 21 control subjects. Athletes had been training for at least 1(1/2) years. MAIN OUTCOME MEASURE: Bone speed of sound (bilateral) at the distal radius and the midtibia. RESULTS: Gymnasts were significantly shorter and lighter than swimmers and control girls (P<.001) but had a body mass index similar to that of swimmers. Adiposity was lower in athletes than in controls. Speed of sound did not correlate with measures of body size. Higher mean +/- SD radial speed of sound values (nondominant side) were observed in gymnasts (3764 +/- 104 m/s; P =.045) than in swimmers and control girls (3732 +/- 99 and 3721 +/- 83 m/s, respectively). Mean +/- SD tibial speed of sound values (nondominant side) were similar in gymnasts and swimmers (3629 +/- 87 and 3619 +/- 78 m/s, respectively) and higher in the athletic groups than in the control group (3516 +/- 127 m/s; P<.001). In all 3 groups, no differences were observed between dominant and nondominant sides in the radii or tibias. CONCLUSIONS: Physical exercise, impact and nonimpact, is related to enhanced bone properties, as measured by quantitative ultrasound. Longitudinal studies using various modes of bone evaluation are necessary to determine the long-term effect of various types of exercise on bone properties. PMID- 12580683 TI - Maternal antibiotics and decreased periventricular leukomalacia in very low-birth weight infants. AB - OBJECTIVE: To investigate the effect of maternal antibiotics, given in the predelivery period, on neonatal outcomes. DESIGN: Retrospective cohort study. SETTING: A single level 3 neonatal intensive care unit. PATIENTS: All infants with birth weights 1500 g or less cared for from July 1994 to July 2000 (n = 834) were included in the study. Mothers were classified as receiving antibiotics if they received any parenteral antibiotics in the predelivery period. Infants whose mothers received antibiotics were compared with infants whose mothers received no antibiotics. MAIN OUTCOME MEASURES: The main outcome variables studied included intraventricular hemorrhage (IVH), cystic periventricular leukomalacia (PVL), sepsis, and mortality. RESULTS: Of 834 mothers, 374 (45%) received antibiotics prior to delivery. On univariate analysis, there were no differences in the relative risk (RR) of mortality (1.26; 95% confidence interval [CI], 0.86-1.79) or grades 3 to 4 IVH (RR, 1.39; 95% CI, 0.82-1.90) between the antibiotics and no antibiotics groups. Infants born to mothers receiving antibiotics had an increased risk of culture-proven sepsis (RR, 1.4; 95% CI, 1.02-1.64) and a decreased risk of cystic PVL (RR, 0.26; 95% CI, 0.09-0.79) compared with infants whose mothers did not receive antibiotics. After controlling for confounding variables, maternal antibiotics were not associated with a decrease in the risk of mortality (adjusted risk [AR], 1.0; 95% CI, 0.5-2.1), grades 3 to 4 IVH (AR, 1.0; 95% CI, 0.5-1.9), or sepsis (AR, 0.9; 95% CI, 0.7-1.4). However, the use of maternal antibiotics was associated with a decreased risk of developing cystic PVL (AR, 0.09; 95% CI, 0.02-0.5). CONCLUSIONS: In our population of very low birth-weight infants, maternal antibiotics were associated with a decreased risk of cystic PVL. Maternal antibiotics do not change the risk of mortality, sepsis, or severe IVH. PMID- 12580684 TI - Use of psychotropic agents in preschool children: associated symptoms, diagnoses, and health care services in a health maintenance organization. AB - BACKGROUND: Recent pharmacoepidemiological reports have contributed to concerns about frequent and perhaps indiscriminate psychopharmacotherapy for very young children. OBJECTIVE: To examine the diagnoses, symptoms, and health care services associated with preschool children receiving psychotropic medication. DESIGN: Population-based pharmacoepidemiological analysis of electronic medical records, paper medical and mental health charts, and pharmacy records from 1997 and 1998. SETTING: A large Pacific Northwest health maintenance organization. PARTICIPANTS: Preschool children receiving psychotropic medication (psychostimulants, antidepressants, neuroleptics, or alpha(2)-adrenergic agonists). MEASURES: Physician-reported mental health diagnoses and related symptoms, functional impairment, family and participant characteristics, and the types and level of associated medical and mental health services. RESULTS: Of 743 preschool children who clinicians identified as having behavioral or emotional problems, 120 (16%) received psychotropic medication; 57 children (48%) were prescribed a stimulant medication only, and 60 (50%) received a diagnosis of attention deficit/hyperactivity disorder. Most children had substantial psychosocial risk factors, including parents with psychiatric or substance abuse problems (71%; n = 85), documented histories of abuse (29%; n = 35), and out-of-home placement (31%; n = 37). Four of 5 children or families (83%; n = 99) received psychosocial services in addition to pharmacotherapy. On average, children received psychotropic medication at least 6 months after initial identification of a behavioral or mental health problem. CONCLUSIONS: Despite commentary by the popular media about widespread psychopharmacotherapy for very young children, such treatment was only infrequently received in this health plan. Most children receiving psychopharmacotherapy had substantial additional risk factors and were receiving psychosocial services for mental health or behavioral management. PMID- 12580685 TI - Urine testing for drugs of abuse: a survey of suburban parent-adolescent dyads. AB - BACKGROUND: The American Academy of Pediatrics is opposed to involuntary diagnostic testing for drugs of abuse. OBJECTIVE: To gather data about attitudes of parents and their teenagers about involuntary drug testing on parental request. DESIGN: Adolescents and their accompanying parents separately answered a printed survey in the offices of their private pediatrician. The survey posed 2 hypothetical questions about urine testing: (1) Do parents have the right to ask a teenager's physician to order a urine test for drugs of abuse without the teenager's knowledge-if the teenager has falling school grades, an uncooperative attitude, and major untruthfulness? (2) In such a case, should the teenager's physician obtain a urine test for drugs on parental request only, without the teenager's consent? RESULTS: A total of 393 paired evaluable surveys were collected: 77.6% from Virginia and 22.4% from Ohio. There were no significant differences in answers between the 2 study sites. Of the students, 85.8% had either an A or a B grade point average. Current marijuana use was unusually low in our teenaged respondents. Of the parents surveyed, 81.7% would want a physician to be able to perform a urine test for drugs of abuse for a problematic teenager without the young person's consent. The answers to the 2 questions about urine drug tests had poor kappa coefficients of agreement between teenagers and parents (0.04 and 0.09, respectively). Reanalysis, using the variables of age, grade point average, and frequency of marijuana smoking, showed little difference in agreement scores. CONCLUSIONS: In the 2 suburban pediatric practices surveyed, parental opinions and expectations were at variance with the American Academy of Pediatrics policy statement on nonconsensual urine drug testing in the presence of clinical problems. Pediatricians need to be conscious of this clinical-ethical dilemma, become familiar with the American Academy of Pediatrics policy on drug testing, and develop their own position and expertise in this area. The dyad method (parent-teenager survey) is novel and improved the methodology of our study. We surveyed middle-class suburban adolescents while previous studies of adolescents surveyed inner-city populations. PMID- 12580686 TI - Parental decision making and the choice of communication modality for the child who is deaf. AB - BACKGROUND: Early intervention decisions for a deaf or hard of hearing child are difficult to make, partly because of the lack of definitive proof of the superiority of any particular communication approach. OBJECTIVE: To compare the relative importance of the child's hearing loss and parental attitudes, beliefs, values, and aspirations in the decision process. METHODS: Eighty-three parents were surveyed about decision factors that may have affected their choice of communication modality, including resource availability, attitudes and beliefs about hearing loss, values, trade-offs, and goals. Parental preference ratings on hypothetical outcomes were also collected in 4 domains: communication, academic performance, social functioning, and emotional well-being. RESULTS: The child's extent of hearing loss was the most influential decision factor (P<.001). Beyond the extent of hearing loss, logistic regression further showed that parental cognitive-attitudinal factors were important in the inclination to favor an oral approach-if they believed that deafness can and should be corrected and if they desired the child to be able to speak (P =.03 and.04, respectively). Technology that aims at improving the child's ability to speak (eg, cochlear implants) had no significant impact on the decision to choose oral only training. CONCLUSIONS: Professionals who work with deaf children and their parents should recognize the presence of many relevant issues beyond the extent of the child's hearing loss. Interventions may be most effective if aimed at balancing parental beliefs and aspirations and audiologic considerations. PMID- 12580687 TI - Infrequent parental monitoring predicts sexually transmitted infections among low income African American female adolescents. AB - OBJECTIVE: To prospectively determine (using an 18-month follow-up period) the association between African American female adolescents' perceptions of parental monitoring and their acquisition of biologically confirmed infection with Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis. DESIGN: A prospective cohort study of 217 African American female adolescents enrolled in the control arm of a randomized trial of a human immunodeficiency virus prevention intervention program. SETTING AND PARTICIPANTS: A volunteer sample of adolescents (aged 14-18 years) recruited from low-income neighborhoods characterized by high rates of unemployment, substance abuse, violence, and sexually transmitted diseases. MAIN OUTCOME MEASURES: Adolescents provided 2 self collected vaginal swab specimens. One was tested for C. trachomatis and N. gonorrhoeae DNA with ligase chain reaction. The other was used to inoculate culture medium for T. vaginalis. Identical assay procedures were repeated at the 6-month, 12-month, and 18-month follow-up intervals. RESULTS: Adjusted odds ratios indicated that adolescents who perceived infrequent parental monitoring at baseline were 1.8 (95% confidence interval, 1.01-3.21) and 2.4 (95% confidence interval, 1.22-4.87) times more likely to acquire chlamydia or trichomoniasis, respectively, compared with their counterparts who perceived greater levels of monitoring. Similarly, adolescents who perceived infrequent parental monitoring were 2.1 (95% confidence interval, 1.16-3.74) times more likely to test positive for a sexually transmitted infection during the course of the 18-month follow-up period. CONCLUSIONS: Adolescents' perceptions of their parental-monitoring levels predicted subsequent acquisition of biologically confirmed chlamydia and trichomoniasis infections. These findings suggest that expanded efforts leading toward effective clinic- and community-based sexually transmitted infection intervention programs involving parents may be warranted. PMID- 12580688 TI - Pediatric "body packing". AB - BACKGROUND: Recent events in the United States have led to increased security at national borders, resulting in an unexpected increase in drug seizures. In response, drug smugglers may begin using children as couriers, including using them as "body packers." OBJECTIVE: To look at the occurrence of body packing, the concealing of contraband within the human body, which is well documented in adults, in the pediatric literature. PATIENT REPORTS: Two cases of pediatric body packing, in boys aged 16 years and 12 years. Patient 1, a 16-year-old boy, presented with findings consistent with opioid intoxication after arriving in the United States on a transcontinental flight. His mental status improved after he received naloxone hydrochloride, and he subsequently confessed to body packing heroin. He was treated with a naloxone infusion and aggressive gastrointestinal decontamination. He ultimately passed 53 packets of heroin, one of which had ruptured. He recovered uneventfully. Patient 2, a 12-year-old boy, presented to the emergency department with rectal bleeding. He had recently arrived in the United States from Europe, and he confessed to body packing heroin. He was treated with whole-bowel irrigation and activated charcoal, and he subsequently passed 84 packets. He also recovered uneventfully. CONCLUSIONS: We report the first 2 cases of body packing in the pediatric literature and review the diagnosis and management of this clinical entity. Pediatricians should be aware that body packing, regrettably, is not confined to the adult population. PMID- 12580689 TI - A randomized controlled trial of the middle and junior high school D.A.R.E. and D.A.R.E. Plus programs. AB - OBJECTIVE: To evaluate the effect of the middle and junior high school Drug Abuse Resistance Education (D.A.R.E.) and D.A.R.E. Plus programs on drug use and violence. DESIGN: Randomized controlled trial of 24 schools, with 3 conditions: D.A.R.E. only, D.A.R.E. Plus, and delayed program control. SETTING: Schools and neighborhoods, primarily in Minneapolis-St Paul. PARTICIPANTS: All seventh-grade students in 24 schools in the academic year 1999-2000 (N = 6237 at baseline, 67.3% were white, and there was 84.0% retention at final follow-up). INTERVENTIONS: The middle and junior high school D.A.R.E. curriculum in the 16 schools that received D.A.R.E. only and D.A.R.E. Plus. In the 8 schoolts that received D.A.R.E. Plus, additional components included a peer-led parental involvement classroom program called "On the VERGE," youth-led extracurricular activities, community adult action teams, and postcard mailings to parents. The interventions were implemented during 2 school years, when the cohort was in the seventh and eighth grades. MAIN OUTCOME MEASURES: Self-reported tobacco, alcohol, and marijuana use; multidrug use; violence; and victimization, assessed at the beginning and end of seventh grade and at the end of eighth grade. Growth curve analytic methods were used to assess changes over time by condition. RESULTS: There were no significant differences between D.A.R.E. only and the controls; significant differences among boys between D.A.R.E. Plus and controls for tobacco, alcohol, and multidrug use and victimization; significant differences among boys between D.A.R.E. Plus and D.A.R.E. only in tobacco use and violence; and no significant behavioral differences among girls. CONCLUSION: D.A.R.E. Plus significantly enhanced the effectiveness of the D.A.R.E. curriculum among boys and was more effective than the delayed program controls, underscoring the potential for multiyear, multicomponent prevention programs and demonstrating sex differences in response to intervention programs. PMID- 12580690 TI - Frequency and intensity of activity of third-grade children in physical education. AB - BACKGROUND: Health recommendations are for preadolescent children to have daily school physical education (PE) classes that engage children in moderate to vigorous physical activity at least 50% of class time. OBJECTIVE: To observe activity of children in PE classes in third grades across 10 different sites. DESIGN: Observational study. SETTING: Six hundred eighty-four elementary schools in 10 sites. SUBJECTS: A total of 814 children (414 boys, 400 girls; mean age, 9.0 years) enrolled in the National Institute of Child Health and Human Development Study of Early Child Care and Youth Development. METHODS: Each child was observed during 1 scheduled PE class. MAIN OUTCOME MEASURE: The SOFIT (System for Observing Fitness Instruction Time) observation method, a validated, heart rate observation system, yields levels of activity the child is engaged in as well as the lesson context, type of teacher, and location of the PE class. RESULTS: Children averaged 2.1 PE lessons per week, of 33 minutes each. Only 5.9% of children had daily PE. Children accrued 4.8 very active and 11.9 minutes of moderate to vigorous physical activity per PE lesson, 15.0% and 37.0% of lesson time, respectively. Lesson length and number of minutes per week were similar for boys and girls; however, boys spent proportionately more PE time in very active and moderate to vigorous activity. This resulted in boys having a higher energy expenditure rate than girls. CONCLUSION: Children observed in this study received 25 min/wk of moderate to vigorous activity in school PE, falling far short of national recommendations. PMID- 12580691 TI - Time spent by primary care practices on pediatric influenza vaccination visits: implications for universal influenza vaccination. AB - OBJECTIVE: To measure the time currently spent by primary care practice personnel, and the examination room occupancy time for childhood influenza vaccination visits, to assess the practicality of annual influenza vaccination of all preschool children. SETTING: Seven primary care practices serving one fourth of the children living in Rochester, NY. PATIENTS: Ninety-two children seen for influenza vaccination visits in the 2000-2001 vaccination season. METHODS: Using a standardized protocol, practice staff measured the time spent on check-in, nurse or physician examination, and the actual influenza vaccination process. Waiting and "hands-on" times were determined, as well as total visit and room occupancy times. Nonparametric tests and multivariable models were used to analyze the time spent for components of the visits and to compare time spent by different age groups and practice types (suburban or urban). RESULTS: The median duration of the influenza vaccination visit was 14 minutes (25th to 75th percentiles range, 9-25 minutes) across the 7 practices, with visits to urban practices being longer (22 minutes) than visits to suburban practices (9 minutes). Eighty percent of patient time involved waiting, primarily in examination rooms. The major components of influenza vaccination visits included waiting room time (4 minutes in suburban practices vs 8 minutes in urban practices; P<.01), and time in the examination room (5 minutes vs 14 minutes, respectively; P<.001), during which only 1 to 2 minutes (for both suburban and urban practices) were for hands-on vaccinations. Only 5% of visits were examined by a physician or nurse practitioner. Visit times did not vary by age. CONCLUSIONS: Although the personnel time for influenza vaccination visits was short, there was substantial patient waiting and long occupancy of examination rooms. If universal influenza vaccination is to be efficiently managed in primary care practices, it may be necessary to implement "vaccination clinics" or sessions in which large numbers of children are scheduled for influenza vaccinations at times when adequate rooms and dedicated nursing staff are available. PMID- 12580692 TI - Child care and common communicable illnesses in children aged 37 to 54 months. AB - OBJECTIVE: To examine the relationship between experience in child care and communicable illnesses (gastrointestinal tract infection, upper respiratory tract infection, and otitis media) in children aged 37 months to 54 months with particular focus on the effect of entry into child care after age 3 years. DESIGN: Health, child care, and family data were obtained from more than 1100 participants in the National Institute of Child Health and Human Development Study of Early Child Care, a 10-site prospective study that began at birth. Longitudinal logistic regression analyses were performed using each type of communicable illness as the outcome variable, with family, child, and child care variables as predictors in the model. RESULTS: For children aged 37 to 54 months, rates of upper respiratory tract illness, gastrointestinal tract illness, and ear infections were higher in those enrolled in child care arrangements with more than 6 children. During this period, children with experience in large-group care prior to age 3 years were less likely to be ill than children who entered child care for the first time after age 3 years. Even so, their rates were still higher than for those in small-group care or who were cared for at home between the ages of 37 and 54 months. Reported rates of respiratory tract and gastrointestinal tract illnesses were higher for European American than African American children. Number of hours per week in child care was not a factor. CONCLUSION: Children in child care arrangements with more than 6 other children experience more bouts of upper respiratory tract illness between the ages of 37 and 54 months. PMID- 12580693 TI - Fever, hepatosplenomegaly, and pancytopenia in a 5-month-old infant. PMID- 12580694 TI - Board preparation strategies. PMID- 12580695 TI - Senior resident autonomy in a pediatric hospitalist system. PMID- 12580696 TI - Palliative program considerations. PMID- 12580698 TI - Apolipoprotein E, Alzheimer disease, and African Americans. PMID- 12580699 TI - Hashimoto encephalopathy: syndrome or myth? AB - BACKGROUND: Hashimoto encephalopathy has been described as a syndrome of encephalopathy and high serum antithyroid antibody concentrations that is responsive to glucocorticoid therapy, but these could be chance associations. OBJECTIVE: To study a patient with Hashimoto encephalopathy and to review the literature to determine whether Hashimoto encephalopathy is an identifiable syndrome. DATA SOURCES AND EXTRACTION: We searched the MEDLINE database to June 2002 for "Hashimoto" or "autoimmune thyroiditis" and "encephalopathy" and examined all identified articles and articles referenced therein, including all languages. We included all patients with noninfectious encephalopathy (clouding of consciousness and impaired cognitive function) and high serum antithyroid antibody concentrations. We excluded patients if they did not meet these inclusion criteria or if their symptoms could be explained by another neurologic disorder. We recorded clinical features and the results of imaging, electroencephalographic, thyroid function, and cerebrospinal fluid studies. DATA SYNTHESIS: We identified 85 patients (69 women and 16 men; mean age, 44 years) with encephalopathy and high serum antithyroid antibody concentrations. Among these patients, 23 (27%) had strokelike signs, 56 (66%) had seizures, 32 (38%) had psychosis, 66 (78%) had a high cerebrospinal fluid protein concentration, and 80 (98%) of 82 had abnormal electroencephalographic findings. Thyroid function varied from overt hypothyroidism to overt hyperthyroidism; the most common abnormality was subclinical hypothyroidism (30 patients [35%]). Among patients treated with glucocorticoids, 66 (96%) improved. CONCLUSIONS: The combination of encephalopathy, high serum antithyroid antibody concentrations, and responsiveness to glucocorticoid therapy seems unlikely to be due to chance. However, there is no evidence of a pathogenic role for the antibodies, which are probably markers of some other autoimmune disorder affecting the brain. PMID- 12580700 TI - The reach of neurology. AB - Neurologists experienced in the interpretation of disease in terms of disordered action of the nervous system should be well suited to extend their field of interest to the more complex disorders of human behavior, including hysteria, delirium, ill-defined pain syndromes, unexplained fatigue, disorders of thought, atypical depression, and delusions. To illustrate the potential of neurology in approaching the more complex disorders of behavior, several examples from clinical neurology are presented in which phenomena calling for inquiry and analysis in neurological terms are described. The categories are temporal lobe epilepsy, delirium, drug toxicity, disease processes of the cerebrum, obscure pain, dyslexia, and hysteria. Inquiry into complex disorders of behavior is inseparable from the broad subject of normal mental activity, the neural organization subserving all human thought, emotion, and action. Because of this close association, the comment on hysteria includes an introduction to the important question of whether we humans possess a free will to choose our course of behavior. PMID- 12580701 TI - Recurrent corticocortical interactions in neural disease. PMID- 12580702 TI - Incidence of Alzheimer disease in a biracial urban community: relation to apolipoprotein E allele status. AB - CONTEXT: Few studies compare Alzheimer disease (AD) incidence among black and white subjects. OBJECTIVE: To estimate incidence and the effect of the apolipoprotein E (APOE) epsilon4 allele in these races. DESIGN: Population-based study of disease incidence using a random, stratified sample. SETTING: A geographically defined community of 3 adjacent neighborhoods in Chicago, Ill. PARTICIPANTS: A total of 6158 persons (78.7% overall; 80.5% of the black subjects and 74.6% of the white subjects) participated; 4.1 years later, persons initially free of AD were sampled for clinical evaluation for disease incidence (overall 842 persons [74.8%] participated; 67.6% of the black subjects and 81.9% of the white subjects). INTERVENTIONS: None. MAIN OUTCOME MEASURE: Incident, clinically diagnosed AD. RESULTS: The effect of the APOE epsilon4 allele on the risk of AD differed strongly for black and white subjects. Among white subjects, the presence of the APOE epsilon4 allele was associated with a 2.73-fold (95% confidence interval [CI], 1.40-5.32) increase in risk while among black subjects there was no increase in risk (odds ratio, 1.02; 95% confidence interval, 0.39 2.68). Black race was associated with a nonsignificantly increased risk of AD with an odds ratio of 1.84 (95% CI, 0.73-4.66) if APOE and its interaction with race are considered, and an odds ratio of 1.28 (95% CI, 0.54-2.98) if they are not. The incidence of AD was 1.45% (95% CI, 0.89%-2.01%) per year among persons 65 to 74 years old, 4.73% (95% CI, 3.83%-5.64%) among those 75 to 84 years old, and 9.11% (95% CI, 7.36%-10.9%) among those 85 years and older. CONCLUSION: Apolipoprotein E epsilon4 led to increased risk of AD among white subjects but not black subjects. PMID- 12580703 TI - Dietary fats and the risk of incident Alzheimer disease. AB - BACKGROUND: Few studies have investigated the effects of dietary fats on the development of Alzheimer disease. We examined the associations between intake of specific types of fat and incident Alzheimer disease in a biracial community study. METHODS: We performed clinical evaluations on a stratified random sample of 815 community residents aged 65 years and older who were unaffected by Alzheimer disease at baseline and who completed a food-frequency questionnaire a mean of 2.3 years before clinical evaluation. RESULTS: After a mean follow-up of 3.9 years, 131 persons developed Alzheimer disease. Intakes of saturated fat and trans-unsaturated fat were positively associated with risk of Alzheimer disease, whereas intakes of omega-6 polyunsaturated fat and monounsaturated fat were inversely associated. Persons in the upper fifth of saturated-fat intake had 2.2 times the risk of incident Alzheimer disease compared with persons in the lowest fifth in a multivariable model adjusted for age, sex, race, education, and apolipoprotein E epsilon4 allele status (95% confidence interval, 1.1-4.7). Risk also increased with consumption of trans-unsaturated fats, beginning with the second fifth of intake (relative risk, 2.4 compared with the lowest fifth; 95% confidence interval, 1.1-5.3). We observed linear inverse associations between Alzheimer disease and vegetable fat (P =.002), and, after further adjustment for other types of fat, marginally significant associations with intake of omega-6 polyunsaturated fat (P =.10 for trend) and monounsaturated fat (P =.10 for trend). Intakes of total fat, animal fat, and dietary cholesterol were not associated with Alzheimer disease. CONCLUSION: High intake of unsaturated, unhydrogenated fats may be protective against Alzheimer disease, whereas intake of saturated or trans-unsaturated (hydrogenated) fats may increase risk. PMID- 12580704 TI - Antioxidant vitamin intake and risk of Alzheimer disease. AB - BACKGROUND: The generation of oxygen free radicals is involved in the pathogenesis of Alzheimer disease (AD). OBJECTIVE: To determine whether the intake of antioxidant vitamins decreases the risk of AD. METHODS: We investigated the relationship between AD and the intake of carotenes, vitamin C, and vitamin E in 980 elderly subjects in the Washington Heights-Inwood Columbia Aging Project who were free of dementia at baseline and were followed for a mean time of 4 years. Semiquantitative food frequency questionnaires were administered between baseline and the first follow-up visit. Cox proportional hazards regression models were conducted with quartiles of each vitamin intake as the exposure of interest and incident AD as the outcome, adjusted for age, level of education, sex, APOE epsilon4 status, ethnicity, and smoking. RESULTS: There were 242 incident cases of AD in 4,023 person-years of follow-up (6 per 100 person-years). Intake of carotenes and vitamin C, or vitamin E in supplemental or dietary (nonsupplemental) form or in both forms, was not related to a decreased risk of AD. Trend tests for the association between quartiles of total intake of vitamins C and E also were not significant. CONCLUSION: Neither dietary, supplemental, nor total intake of carotenes and vitamins C and E was associated with a decreased risk of AD in this study. PMID- 12580705 TI - Estrogen levels do not correlate with improvement in cognition. AB - OBJECTIVE: To investigate whether an association exists between estradiol and estrone levels and measures of cognitive functioning in women with Alzheimer disease (AD) treated with conjugated equine estrogen (Premarin; Wyeth-Ayerst, Philadelphia, Pa). METHODS: We studied 120 postmenopausal women who underwent hysterectomy and who had AD treated with Premarin for 1 year. Plasma estradiol and estrone levels were determined at multiple points during the 1-year treatment trial. The change from baseline level at 2 and 12 months was associated with the change score on 7 different assessments of cognitive functioning. RESULTS: At baseline, estradiol levels were low and there were no associations between the estradiol level and the 7 neuropsychological measures. A similar pattern was observed for estrone treatment. During treatment with 0.625 mg/d of Premarin, estradiol levels increased about 4-fold; while receiving 1.25 mg/d of Premarin, estradiol levels increased about 8-fold. A similar pattern was seen with estrone treatment. For both estradiol and estrone levels, there were no significant associations between the change in plasma level and the change in neuropsychological test scores at either 2 or 12 months. CONCLUSION: Although Premarin elevated estradiol and estrone levels, there was no association between hormone levels and cognitive functioning after either 2 or 12 months of treatment. PMID- 12580706 TI - Higher estrogen levels are not associated with larger hippocampi and better memory performance. AB - BACKGROUND: Estrogens may prevent cognitive decline and Alzheimer disease. Animal study findings have shown beneficial effects of estrogen on the brain, particularly on the hippocampus, a structure related to memory performance and early Alzheimer disease. OBJECTIVE: To investigate whether higher levels of endogenous estradiol in older women and men are associated with larger hippocampal volumes on magnetic resonance imaging and better memory performance. DESIGN AND SETTING: Cross-sectional analysis within the Rotterdam Scan Study, a population-based study in the Netherlands of elderly subjects who do not have dementia. PARTICIPANTS: Two hundred ten women and 202 men, aged 60 to 90 years, with plasma levels of total estradiol and, in part, 162 women and 149 men also with levels of bioavailable and free estradiol. MAIN OUTCOME MEASURE: Hippocampal volumes on magnetic resonance imaging and memory performance (delayed recall). RESULTS: Women with higher total estradiol levels had smaller hippocampal volumes and poorer memory performance -0.29 mL (95% confidence interval, -0.57 to -0.00) and -0.4 (95% confidence interval, -1.3 to 0.5) fewer words in delayed recall testing for the highest tertile compared with the lowest tertile. Similar inverse associations were found among bioavailable and free estradiol levels, hippocampal volumes, and memory. In men, no association was observed between estradiol levels and hippocampal volume, but a trend was found for higher levels of total estradiol to be associated with poorer memory performance. CONCLUSION: Our data do not support the hypothesis that higher levels of endogenous estradiol in older women and men are associated with larger hippocampal volumes and better memory performance. PMID- 12580707 TI - Low blood pressure and risk of dementia in the Kungsholmen project: a 6-year follow-up study. AB - BACKGROUND: Previous studies have reported a higher prevalence of dementia in persons with low blood pressure. OBJECTIVE: To examine whether low blood pressure is prospectively associated with the occurrence of Alzheimer disease and dementia in elderly people. SUBJECTS AND METHODS: A community-based, dementia-free cohort (n = 1270) aged 75 to 101 years was longitudinally examined twice within 6 years to detect incident dementia using the Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition criteria. Cox proportional hazards models were used to analyze blood pressure in association with dementia after adjustment for several potential confounders. RESULTS: During the 6-year period, 339 subjects were diagnosed with dementia, including 256 persons with Alzheimer disease. Subjects with very high systolic pressure (>180 vs 141-180 mm Hg) had an adjusted relative risk of 1.5 (95% confidence interval [CI], 1.0-2.3; P =.07) for Alzheimer disease, and 1.6 (95% CI, 1.1-2.2) for dementia. Low systolic pressure (90 mm Hg) was not associated with dementia incidence, whereas extremely low diastolic pressure ( or = 100 microg/L), mild (50-99 microg/L) or moderate-severe (< 50 microg/L), and classified according to sex, school year and presence of goitre. RESULTS: 76% (439/577) of students had abnormal urinary iodine values, with 27% (156/577) having values consistent with moderate-severe deficiency. The median urinary iodine excretion for the total group was 70 microg/L, with values for school years 5-12 ranging from 62 microg/L (Year 12) to 76 microg/L (Year 9). The median urinary iodine value in girls was lower than that in boys (64 microg/L v 82 microg/L), and girls had significantly lower urinary iodine values overall (P < 0.002). There was no association between goitre grade and moderate-severe (< 50 microg/L; P = 0.39) or mild (50-99 microg/L; P = 0.07) urinary iodine deficiency. CONCLUSIONS: We found mild iodine deficiency in a cohort of schoolchildren in Melbourne. Our results support other data showing mild iodine deficiency in Sydney and Tasmania and the argument for a national study of iodine nutrition. PMID- 12580742 TI - Preparedness for hospital practice among graduates of a problem-based, graduate entry medical program. AB - OBJECTIVE: To compare preparedness for hospital practice between graduates from a problem-based, graduate-entry medical program and those from other programs (undergraduate problem-based and traditional). DESIGN: Survey of graduates (by mailed questionnaire) and organisers of clinical training (by semistructured interview); results were compared with published results of surveys of graduates from other programs. SETTING AND PARTICIPANTS: All graduates of the first intake of the University of Sydney graduate-entry medical program were surveyed at the end of their first intern year (2001), along with the director of clinical training or intern manager at each of the New South Wales hospitals that employed the graduates. MAIN OUTCOME MEASURES: Graduates' self-reported level of preparedness in the eight domains of the Preparation for Hospital Practice Questionnaire; and organisers' opinions of their strengths and weaknesses. RESULTS: 76 of 108 graduates from the graduate-entry program (70%) and organisers of clinical training at all 17 hospitals participated. Graduates from the program felt more prepared than did those from other programs in five of the eight domains assessed (interpersonal skills, confidence, collaboration, holistic care, and self-directed learning) and no less prepared in any domain. Organisers rated the graduates highly, especially in clinical competence, confidence, communication and professional skills. Opinions of interns' knowledge of basic sciences conflicted, with strengths and weaknesses mentioned with equal frequency. CONCLUSION: Graduates from the graduate-entry, problem-based program are at least as well prepared for their intern year as graduates from traditional and undergraduate problem-based programs. PMID- 12580743 TI - Medical students and clinical ethics. PMID- 12580744 TI - A week at the morgue: a personal odyssey. PMID- 12580745 TI - "Death talk": debating euthanasia and physician-assisted suicide in Australia. AB - Imprecise language and deliberate confusion of important ethical and legal concepts are clouding our understanding of controversial end-of-life issues. This could affect our decision about whether or not to legalise euthanasia. PMID- 12580746 TI - Childhood tetanus in Australia: ethical issues for a should-be-forgotten preventable disease. AB - Refusal of a parent to have a child vaccinated against tetanus raised ethical issues for the treating clinicians. The clinicians felt their duty to the child was compromised, but recognised that our society leaves the authority for such decisions with the parents. As there was no reason, other than different beliefs about vaccination, to doubt the parent's care for the child, the clinicians limited their response to providing strong recommendations in favour of vaccination. Other issues raised by this case include community protection, and the costs to the community of treating a vaccine-preventable disease. PMID- 12580747 TI - Medical history and medical practice: persistent myths about the foreskin. AB - Although many 19th-century misconceptions about the foreskin have been dispelled since it was shown that infantile phimosis was not an abnormality, the ideas that ritual or religious circumcision arose as a hygiene measure, and that circumcision makes no difference to sexual response, have persisted. The first idea should be dismissed as a myth and the second has been seriously questioned by modern research. PMID- 12580748 TI - Does a combined program of dietary modification and physical activity or the use of metformin reduce the conversion from impaired glucose tolerance to type 2 diabetes? PMID- 12580749 TI - Statistical methods in clinical trials. PMID- 12580750 TI - National ethics committee urgently needed. PMID- 12580751 TI - Inappropriate use of hospital emergency departments. PMID- 12580752 TI - Epidemiological modelling (including economic modelling) and its role in preventive drug therapy. PMID- 12580753 TI - Screening mammography and mortality. PMID- 12580754 TI - The Australian Health Care Agreements 2003-2008: reform or false dawn? PMID- 12580755 TI - Pertussis: adults as a source in healthcare settings. PMID- 12580756 TI - Boundaries of medicine. PMID- 12580757 TI - Comparison of the effect of oestradiol, tamoxifen and raloxifene on nerve growth factor-alpha expression in specific neonatal mouse uterine cell types using laser capture microdissection. AB - Oral dosing of CD-1 mice on days 2-5 after birth with tamoxifen but not raloxifene disrupts the development of the myometrium, resulting in adult uterine adenomyosis. Using laser capture microdissection and RT-PCR we have investigated nerve growth factor (NGF) and cognate receptor expression in uterine cells of 6 day-old pups that may be important in early developmental changes that give rise to adenomyosis. NGF down-regulation is known to occur during terminal myogenic differentiation. NGF was found exclusively in endometrial luminal epithelium of controls. It was up-regulated 18-fold in the luminal epithelium following dosing with tamoxifen but not raloxifene. Western blotting for NGF protein in the whole uterus showed a 25-fold increase after tamoxifen treatment. Expression of the low affinity p75 neutrophin receptor (p75(NTR)) was twofold higher in the myometrium compared with luminal epithelium or stroma. This was not altered following tamoxifen treatment. There was no detectable expression of high affinity tyrosine kinase receptor (trkA(NGFR)). This study shows luminal epithelial cells of the endometrium primarily form NGF. This suggests that NGF normally regulates the differentiation of the mesenchyme into uterine myocytes through paracrine mechanisms and that an early disturbance of this process plays a key role in the subsequent development of adenomyosis. PMID- 12580758 TI - Putative functional characteristics of human estrogen receptor-beta isoforms. AB - Estrogen receptors (ERalpha and ERbeta) are clearly multifaceted in terms of structure and function. Several relatively abundant ERbeta isoforms have been identified, which can be differentially expressed in various tIssues. In order to provide insight into the possible role of the ERbeta family in breast tIssue a study of the putative functions of the human (h) ERbeta1, hERbeta2 and hERbeta5 isoforms was undertaken. Only hERbeta1 was found to bind ligand, which induced conformational changes as determined by protease digestion assays. All ERbeta isoforms could bind to and bend DNA although the relative efficiency with which they bound DNA differed with hERalpha>hERbeta1>hERbeta2>>hERbeta5. All ERbeta isoforms inhibited ERalpha transcriptional activity on an estrogen-response element (ERE)-reporter gene. The relative activities were hERbeta1>hERbeta2>hERbeta5; however, only hERbeta1 had transcriptional activity of its own. Both LY117018-hERalpha and LY117018-hERbeta1 complexes alone could activate transcription on a TGF-beta3-CAT gene. Although hERbeta2 and hERbeta5 had no activity alone, they inhibited ERalpha but not hERbeta1 transcriptional activity of transforming growth factor (TGF)-beta3-CAT. In marked contrast to activity on an ERE-CAT reporter gene, hERbeta1 did not modulate ERalpha transcriptional activity on a TGF-beta3-CAT reporter gene. These data support promoter-specific differential activities of hERbeta isoforms with respect to models of ERalpha regulated gene expression, and suggest that they may have a role in differentially modulating estrogen action. PMID- 12580759 TI - Alternative splicing to exon 11 of human prolactin receptor gene results in multiple isoforms including a secreted prolactin-binding protein. AB - Endocrine and autocrine prolactin (PRL) exerts effects on normal breast and breast cancer cells, and high serum PRL is a poor prognostic factor for colorectal cancer. Here we tested the hypothesis that short isoforms of the PRL receptor (PRLR) in human tissue regulate the actions of PRL in cancer. Using 3' RACE we isolated five splice variants of the human PRLR (hPRLR), three of which encode the complete extracellular binding domain. Two of these isoforms, short form 1a (SF1a) and short form 1b (SF1b), possess unique intracellular domains encoded by splicing to exon 11 from exons 10 and 9 respectively. A third novel isoform (delta7/11) reflects alternative splicing from exon 7 to exon 11 and encodes a secreted soluble PRL-binding protein. Additional splice variants of SF1b and delta7/11 that lacked exon 4 (delta4-SF1b and delta4-delta7/11) were also identified. Functional analyses indicated that hPRLR-SF1b is a strong dominant-negative to the differentiative function of the PRLR long form while hPRLR-SF1a is a weaker dominant-negative. Differential abundance of SF1a, SF1b and delta7/11 expression was detected in normal breast, colon, placenta, kidney, liver, ovary and pancreas, and breast and colon tumors. Taken together, these data indicate the presence of multiple isoforms of the hPRLR that may function to modulate the endocrine and autocrine effects of PRL in normal human tissue and cancer. PMID- 12580760 TI - 1,25-Dihydroxyvitamin D3 inhibits thyroid hormone-induced osteocalcin expression in mouse osteoblast-like cells via a thyroid hormone response element. AB - Thyroid hormones are important regulators of bone development and metabolism. We have demonstrated that tri-iodothyronine (T3) increased and 1,25-dihydroxyvitamin D3 (1,25D3) attenuated the T3-stimulated expression of osteocalcin (OCN) in the osteoblast-like cell line MC3T3-E1. By means of transfection of promoter-reporter gene constructs we investigated the basal and the regulated transcription of this gene by both hormones. We found that a 0.67 kbp and a 1.3 kbp fragment of the mouse OCN OG2 promoter containing two Runx2 binding sites were significantly more active than a smaller fragment containing only one Runx2 binding site. The longer promoter fragments showed a higher reporter gene expression when the transfected cells were treated with 10(-7) M T3. This expression was attenuated by 1,25D3 dose-dependently. These fragments contain a sequence homologue to the recently identified binding site for the 1,25D3 receptor (VDR) in the rat OCN promoter. Deletion of a part of the promoter containing this VDR response element-like sequence (VDRE) resulted in a higher basal expression but abrogated the regulation by T3 and 1,25D3. Electrophoretic mobility shift assays revealed that the deleted sequence was able to bind both in vitro-translated chicken thyroid hormone receptor (TR) and proteins from nuclear extracts that reacted with an antiserum against TR. From these data we conclude that the VDRE-like sequence of the OG2 promoter contains a thyroid hormone response element. PMID- 12580761 TI - Interleukin-1alpha stimulates steroidogenic acute regulatory protein expression via p38 MAP kinase in immature rat Leydig cells. AB - We have investigated the involvement of the steroidogenic acute regulatory (StAR) protein in interleukin-1alpha (IL-1alpha)-induced steroidogenesis in immature (40 day-old) and adult Leydig cells in vitro. Further, IL-1alpha-mediated signaling pathway(s) controlling StAR expression in immature Leydig cells were also studied. IL-1alpha stimulated both androgen production and StAR protein expression in a dose- and time-dependent manner in immature but not adult Leydig cells. These effects of IL-1alpha were prevented by pretreatment of the cells with the specific inhibitors of the p38 MAP kinase, SB203580 and PD169316, suggesting that this kinase is an important part of IL-1alpha signaling in the immature Leydig cell. The present results suggest that IL-1alpha, which is constitutively produced by the rat testis from postnatal day 25, is an important paracrine regulator of postnatal Leydig cell maturation. Regulation of StAR protein expression is one of the possible mechanisms by which IL-1alpha contributes to the differentiation of immature Leydig cells into adult cells. PMID- 12580762 TI - Transient expression pattern of prolactin in Sparus aurata. AB - Using reverse transcription-polymerase chain reaction and in situ hybridization, the expression of the prolactin (PRL) gene was determined during development in gilthead sea bream (Sparus aurata) for the first time. The mRNA for PRL was detected from the second day of the larval stage onwards. This transcript was also located in the adenohypophysial cells, starting at four days post-hatching and was found to be pituitary-specific. Moreover, the possible involvement of PRL in asynchronous growth in the cultivation of gilthead sea bream was also examined. No differences in the distribution of PRL cells were observed in the three sizes of juvenile gilthead sea bream studied. These results suggest that the transcription of PRL is involved in the early development stages of sea bream and that the asynchronous growth-related changes are not due to distinct distribution of PRL cells. PMID- 12580763 TI - Beyond lysis: how complement influences cell fate. AB - Complement is a central component of the innate immune system involved in protection against pathogens. For many years, complement has been known to cause death of targets, either indirectly by attracting and activating phagocytes or directly by formation of a membrane pore, the membrane attack complex. More recently, it has been recognized that complement may cause other 'non-classical' effects that may not directly be aimed at killing of pathogens. Products of complement activation collaborate with the adaptive immune system to enhance responses to antigens. The membrane attack complex of complement, apart from lysing cells, can also trigger diverse events in target cells that include cell activation, proliferation, resistance to subsequent complement attack and either resistance to, or induction of, apoptosis. Various complement products play important roles in signalling for clearance by phagocytes of apoptotic self cells. Here we review some of these non-classical activities of complement and stress the roles that they may play in maintaining the integrity of the organism. PMID- 12580764 TI - Preservation of rat skeletal muscle function during storage for 16 h at 4 degrees C is not improved by pre-storage perfusion. AB - The aim of this study was to investigate if the preservation of isolated skeletal muscles for 16 h at 4 degrees C could be improved by pre-storage perfusion (PSP). Two rat muscle models were used: the soleus (SOL) and a posterior strip of the cutaneous trunci (CT). The effects of a 10 min PSP (at 25 degrees C) with University of Wisconsin solution (UW) or HTK-Bretschneider solution (HTK) on muscle function were analysed. The perfusion model was validated by the demonstration that the SOL and CT could be perfused with donor blood, UW and HTK at a flow rate of 0.2 ml x min(-1) x g(-1) muscle for 10 min without any immediate adverse effects on muscle weight, function (maximum tetanus tension) and cytoarchitecture (multivariate analysis of variance, P >0.05; n =6). For each muscle type and for each solution, six perfused and six non-perfused muscles were stored for 16 h at 4 degrees C. In the perfused groups, the storage and perfusion solution were matched. For both muscle types, the function (maximum tetanus tension), weight and cytoarchitecture of pre-storage perfused muscles was not preserved any better than that of non-perfused muscles, irrespective of the solution used (multivariate analysis of variance, P >0.05). We conclude that PSP for 10 min with UW and HTK does not improve the preservation of function of rat skeletal muscles during storage for 16 h at 4 degrees C. PMID- 12580765 TI - Effects of incremental levels of continuous positive airway pressure on cerebral blood flow velocity in healthy adult humans. AB - Sleep disordered breathing is common in patients with cerebrovascular disease, and could exacerbate the cerebral damage in acute stroke. Data about the effects of continuous positive airway pressure (CPAP) upon cerebral perfusion are conflicting. We investigated whether increasing levels of CPAP may affect cerebral haemodynamics, assessed by transcranial Doppler (TCD) in normal humans. A group of 25 healthy young volunteers were evaluated before (CPAP0-pre), during (CPAP5, CPAP10 and CPAP15, denoting CPAP at 5, 10 and 15 cmH(2)O respectively) and after (CPAP0-post) application of incremental levels of CPAP delivered through a mouthpiece. The mean cerebral blood flow velocity (CBFV) and the pulsatility index (PI; an indirect measure of cerebrovascular resistance) in the middle cerebral artery were measured with TCD. Respiratory rate, heart rate, end tidal carbon dioxide pressure (PETCO(2)), transcutaneous haemoglobin oxygen saturation (SpO(2)), mean arterial blood pressure and anxiety score were also recorded. Compared with CPAP0-pre, CBFV was significantly decreased as higher levels of CPAP were applied (P <0.0001). CPAP15 increased PI (P <0.05), ETCO(2) was reduced by CPAP10 and CPAP15 (P <0.0001), and anxiety score and SpO(2) increased at all levels of CPAP (P <0.05). Heart rate, respiratory rate and mean arterial pressure did not change. The decrease in CBFV was correlated with the fall in P ETCO(2) (CPAP15) and the increase in PI (CPAP10, CPAP15) (P <0.05). In conclusion, even low levels of CPAP delivered through a mouthpiece in awake, young volunteers led to a decrease in CBFV, measured by TCD. This fall in CBFV was associated with hypocapnia and with an increase in both cerebrovascular resistance and anxiety due to breathing against positive pressure. As the negative consequences of a fall in CBFV may outweigh the therapeutic effects of CPAP in the post-stroke setting, further studies of the cerebrovascular effects of CPAP with different interfaces in elderly patients with and without stroke are needed before intervention trials can be performed safely. PMID- 12580766 TI - Hormone-induced modifications of the chromatin structure surrounding upstream regulatory regions conserved between the mouse and rabbit whey acidic protein genes. AB - The upstream regulatory regions of the mouse and rabbit whey acidic protein (WAP) genes have been used extensively to target the efficient expression of foreign genes into the mammary gland of transgenic animals. Therefore both regions have been studied to elucidate fully the mechanisms controlling WAP gene expression. Three DNase I-hypersensitive sites (HSS0, HSS1 and HSS2) have been described upstream of the rabbit WAP gene in the lactating mammary gland and correspond to important regulatory regions. These sites are surrounded by variable chromatin structures during mammary-gland development. In the present study, we describe the upstream sequence of the mouse WAP gene. Analysis of genomic sequences shows that the mouse WAP gene is situated between two widely expressed genes (Cpr2 and Ramp3). We show that the hypersensitive sites found upstream of the rabbit WAP gene are also detected in the mouse WAP gene. Further, they encompass functional signal transducer and activator of transcription 5-binding sites, as has been observed in the rabbit. A new hypersensitive site (HSS3), not specific to the mammary gland, was mapped 8 kb upstream of the rabbit WAP gene. Unlike the three HSSs described above, HSS3 is also detected in the liver, but similar to HSS1, it does not depend on lactogenic hormone treatments during cell culture. The region surrounding HSS3 encompasses a potential matrix attachment region, which is also conserved upstream of the mouse WAP gene and contains a functional transcription factor Ets-1 (E26 transformation-specific-1)-binding site. Finally, we demonstrate for the first time that variations in the chromatin structure are dependent on prolactin alone. PMID- 12580771 TI - Delivery of a growth factor fusion protein having collagen-binding activity to wound tissues. AB - Recently, we established a collagen-binding growth factor consisting of epidermal growth factor and the fibronectin collagen-binding domain (FNCBD-EGF). FNCBD-EGF is a biologically active fusion protein that could stably bind to collagen materials, and exert its growth factor activity even after collagen binding. In this study, we investigated the concept that FNCBD moiety with high collagen affinity may enhance the effective local concentration of EGF at the site of administration in the following tissues: skin wounds, catheter-injured arteries, and hind limb muscles. In an animal model of impaired wound healing, application of FNCBD-EGF in combination with collagen gel induced granulation tissue formation in the wounds due to its sustained retention. In the injured artery, infused FNCBD-EGF remained bound to collagen exposed on the injured tissues even after blood circulation was restored. Injection of the fusion protein into the hind limbs revealed that our delivery system was effective for direct administration to muscular tissue. PMID- 12580772 TI - Growth of human endothelial cells on different concentrations of Gly-Arg-Gly-Asp grafted chitosan surface. AB - To improve the adhesion and growth of endothelial cells on chitosan, different concentrations of cell adhesive peptide, Gly-Arg-Gly-Asp (GRGD), were photochemically grafted to its surface. After different concentrations of GRGD SANPAH (N-Succinimidyl-6-4'-azido-2'-nitrophenylamino]-hexanoate) solutions were formed, they were grafted to chitosan surfaces by performing surface adsorption of the peptide solutions and subsequent ultraviolet (UV) irradiation to induce photoreaction. The grafted efficiency for GRGD peptide to chitosan surface was higher than 80%, as semiquantitatively analyzed by a high performance liquid chromatography (HPLC). FTIR spectra and electron spectroscopy for chemical analysis (ESCA) also confirmed that GRGD was successfully grafted to form chitosan-GRGD surfaces. For example, the composition fraction of nitrogen (N1s) for chitosan-GRGD surface grafted at 0.05 M GRGD was 6.8% instead of 3.2% for chitosan only. Human umbilical vein endothelial cells (HUVECs) adhered well and grew on different GRGD grafted concentrations of chitosan-GRGD surfaces after 36 hrs of incubation, but not on the chitosan surface. Furthermore, viability for the growth of HUVECs was increased with an increase in initial concentration of GRGD grafted chitosan-GRGD surfaces analyzed by MTT assay. In conclusion, an increased concentration of GRGD grafted on the chitosan-GRGD surface leads to more HUVECs adhered and grown on the surface. PMID- 12580773 TI - Collagen-hydroxyapatite microspheres as carriers for bone morphogenic protein-4. AB - The purpose of the current study is to evaluate the carrier capability of collagen-hydroxyapatite microspheres to the bone morphogenic proteins (BMP). After anesthesia, a bone defect (6.0 mm in diameter and 10.0 mm in depth) was created at the distal femoral condyles of New Zealand white rabbits. These 10.0 mm3 defects were then completely filled with the implant materials. After 2, 4, 6, and 8 weeks, the animals were sacrificed and histological evaluations were performed. The results showed that when the defects were left untreated, there was no evidence of bone formation during the eight-week experimental period. In the group treated with collagen-hydroxyapatite microspheres without BMP-4, the defect was filled with fibrous tissue and inflammatory cells, while active bone formation with mature marrow tissue formation was evident in the defect treated with collagen-hydroxyapatite microspheres containing BMP-4. Collagen hydroxyapatite microspheres were expected to be replaced by the regenerated bone structure as the bone reconstruction and bone remodelling process occurred. It was apparent that bone regeneration was influenced by the addition of BMP-4. Collagen-hydroxyapatite microspheres were good carriers for bone morphogenic proteins. PMID- 12580774 TI - Investigation of materials for blood-immersed bearings in a microaxial blood pump. AB - Rotary blood pumps are gaining popularity among cardiothoracic surgeons. This article presents an in vitro investigation for choosing a suitable mechanical bearing system in a medium-long term microaxial pump. Different metallic, polymeric, and ceramic components are introduced. Polymers displayed mechanical insufficiency for the application, whereas certain ceramics displayed an inconsistent pattern of failure. We are in search of a compromise in properties that would favor a durable material combination. PMID- 12580775 TI - Safety and efficacy of off-pump coronary artery bypass grafting in chronic dialysis patients. AB - Off-pump coronary artery bypass grafting (CABG) has been recently revived, because cardiopulmonary bypass (CPB) appears to worsen the multiple organ dysfunction after conventional CABG. To evaluate the safety and efficacy of the off-pump CABG in chronic dialysis patients, we compared the perioperative morbidity and mortality between 15 dialysis patients who underwent off-pump CABG at our center over the past 8 years with that of a concurrent group of 19 patients who underwent conventional CABG. Patients were selected for off-pump CABG only when complete revascularization was technically feasible. We found that off-pump CABG is as safe and effective as conventional CABG in selected dialysis patients. It might even be beneficial, because it is associated with less hematocrit drop and blood product use, a lower catabolic rate, and fewer dialysis requirements after surgery. However, the impact of off-pump technique on the long term clinical outcome and resource utilization in renal patients requires further investigation. PMID- 12580776 TI - Improved therapeutic efficacy in cardiomyocyte transplantation for myocardial infarction with release system of basic fibroblast growth factor. AB - Several studies have demonstrated that cell transplantation was effective for the therapy of myocardial infarction. However, little care has been taken for the blood supply indispensable to cell transplantation. This study is an investigation to evaluate the feasibility of in advance angiogenesis by gelatin microspheres incorporating basic fibroblast growth factor (bFGF) for the transplantation therapy of cardiomyocytes with an ischemic cardiomyopathy model. Rats with myocardial infarction received the intramuscular injection of culture medium (Control), or that containing fetal cardiomyocytes (TX) or gelatin microspheres incorporating bFGF (FGF), and gelatin microspheres incorporating bFGF plus fetal cardiomyocytes 1 week later (FGF-TX). The left ventricle (LV) function of rat hearts was assessed by echocardiography and cardiac catheterization 4 weeks later. The LV maximum time-varying elastance was significantly higher in the FGF-TX group than in other groups. The combination of bFGF-induced angiogenesis and cardiomyocyte transplantation is a promising procedure to improve the LV function in rats with myocardial infarction. PMID- 12580777 TI - Development of an implantable high-energy and compact battery system for artificial heart. AB - The purpose of this study was to demonstrate the feasibility of the use of an implantable, high-energy, and compact battery system for an undulation pump total artificial heart (UPTAH). The implantable battery system tested consists of six lithium-ion batteries in series, a charge unit, and a charge/discharge control unit. A lithium-ion battery is currently the best energy-storage device because it has more energy density, a better life cycle, and a smaller temperature rise than those of other secondary batteries. The performance of the implantable battery system was evaluated in an in vitro experiment using an electric load that simulated the UPTAH. Also, sufficiently reliable operation of a system for supplying energy to a UPTAH consisting of a transcutaneous energy transmission system (TETS) and an implantable battery system was confirmed in three experiments using goats. The results of the in vitro and in vivo experiments showed that the implantable battery system supplied sufficient current to the UPTAH for maintenance of physiological conditions in the goat with maximum rise in temperature to less than 43 degrees C. PMID- 12580778 TI - In vivo evaluation of a MPC polymer coated continuous flow left ventricular assist system. AB - The aim of this study was the evaluation of the thrombogenicity and the biocompatibility of the SunMedical EVAHEART left ventricular assist system (LVAS) coated with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer compared to a diamond-like carbon (DLC) coating. Four calves were implanted with the MPC polymer-coated LVAS. Eight calves were implanted with DLC coated LVAS. The thrombogenicity and biocompatibility of the pumps were evaluated. At explant, 60.0 +/- 37.2% (5-85%) of the pump surface area was still coated with MPC polymer after the duration of 45.0 +/- 32.0 days. In 1 out of 4 MPC and 2 out of 8 DLC coated pumps, there was a very small amount of thrombus around the seal ring; otherwise the blood contacting surfaces were free of thrombus. Major organs were normal except for a few lesions in kidneys from both groups. The MPC polymer coated EVAHEART LVAS seems to have low thrombogenicity and high biocompatibility similar to the DLC coated system. The current study demonstrated that the MPC polymer coating shows great promise for being used as an antithrombogenic substrate for the LVAS due to its ease of application, significant cost benefit, and reduction in anticoagulation therapy in acute postoperative period. PMID- 12580779 TI - Development of disposable self-regulating blood pumps and automatically controlled portable extracorporeal membrane oxygenation systems for neonatal extracorporeal membrane oxygenation. AB - For safer and simpler neonatal ECMO management, the authors have been developing an innovative, automatically-controlled ECMO machine using self-regulating blood pumps. The newest model is an air-driven, tube-type blood pump. A pair of blood pumps are placed in parallel and actuated alternately with compressed air. The pump flow is automatically regulated in accordance with hemodynamic changes of the body. The need for a venous reservoir is eliminated because the circuit does not generate excessive negative pressure when there are shortages of draining blood. Therefore, the priming volume of the circuit is only 85 ml. This ECMO apparatus has three driving modes, one of which is a "delay" mode. This enables the pump flow to easily be varied from 10 to 500 ml/min under automatic control, and it can be used when the pump flow is decreased for weaning. This newest ECMO apparatus was clinically used in 3 neonates with severe lung hypoplasia and persistent fetal circulation. The ECMO duration was 139 h to 168 h, and the maximum ECMO flow 71.3 to 109.0 ml/min/kg. Thanks to the self-regulation, intensive observation of the circuit was not necessary during ECMO. Damage to blood cells was less significant, and the use of blood products was only minimal (15.7 ml/kg/day) because a large volume of blood to stabilize the pump performance was not necessary. In conclusion, the automatically-controlled ECMO apparatus worked well without complications by changing the ECMO flow automatically in accordance with hemodynamic changes. This automatic ECMO system required fewer personnel expenditures and was also favorable in terms of hematological findings. PMID- 12580781 TI - Putative in silico mapping of DNA sequences to livestock genome maps using SSLP flanking sequences. AB - In this study, an in silico approach was developed to identify homologies existing between livestock microsatellite flanking sequences and GenBank nucleotide sequences. Initially, 1955 bovine, 1570 porcine and 1121 chicken microsatellites were downloaded and the flanking sequences were compared with the nr and dbEST databases of GenBank. A total of 74 bovine, 44 porcine and 37 chicken microsatellite flanking sequences passed our criteria and had at least one significant match to human genomic sequence, genes/expressed sequence tags (ESTs) or both. GenBank annotation and BLAT searches of the UCSC human genome assembly revealed that 38 bovine, 13 porcine and 17 chicken microsatellite flanking sequences were highly similar to known human genes. Map locations were available for 67 bovine, 44 porcine and 21 chicken microsatellite flanking sequences, providing useful links in the comparative maps of humans and livestock. In support of our approach, 112 alignments with both microsatellite and match mapping information were located in the expected chromosomal regions based on previously reported syntenic relationships. The development of this in silico mapping approach has significantly increased the number of genes and EST sequences anchored to the bovine, porcine and chicken genome maps and the number of links in various human-livestock comparative maps. PMID- 12580780 TI - Polymorphisms in MHC-DRA and -DRB alleles of water buffalo (Bubalus bubalis) reveal different features from cattle DR alleles. AB - Seventy-five individuals of Bubalus bubalis belonging to four different breeds, three of river buffalo and one of swamp buffalo, were studied for polymorphism in MHC DRB (Bubu-DRB) and DRA (Bubu-DRA) loci. Eight alleles of Bubu-DRB were found, and all alleles in the swamp type were shared with the three river breeds. All alleles sampled from the breed of European origin (Mediterranean) were present in breeds sampled in Brazil, thus variability of this locus may have been preserved to a great extent in the more recently founded Brazilian population. Bubu-DRB alleles contained higher proportions of synonymous vs. non-synonymous substitutions in the non-peptide-binding sites (PBS) region, in contrast to the pattern of variation found in BoLA-DRB3, the orthologous locus in cattle. This indicated that either the first domain exon (exon 2) of Bubu-DRB has not undergone as much recombination and/or gene conversion as in cattle alleles, or Bubu-DRB may be more ancient than BoLA-DRB3 alleles. Phylogenetic analysis of DRB alleles from Bubalus, Syncerus c. caffer, the Cape buffalo, and domestic cattle demonstrated transspecies polymorphism. Water buffalo contained two alleles of DRA that differed from each other in two amino acid positions, including one in the PBS (alpha22) that was also shared with Anoa depressicornis, the anoa. Discovery of variation in DRA was surprising as the first domain of DRA is a highly conserved polypeptide in mammals in general and especially in ruminants, where no other substitution in PBS was seen. PMID- 12580782 TI - Polymorphism at the ovine beta3-adrenergic receptor locus: associations with birth weight, growth rate, carcass composition and cold survival. AB - The beta3-adrenergic receptors (ADRB3s) are predominantly found on the surface of adipocytes and are the major mediators of the lipolytic and thermogenic effects of high catecholamine concentrations. Polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis of part of the ovine beta3 adrenergic receptor gene (ADRB3) intron was used to screen 12 large Merino half sib families for sequence variation. Six different alleles that segregated in a Mendelian fashion were observed. The genetic basis for the allelic differences were identified by sequencing the ADRB3 (coding and non-coding regions) from animals that were homozygous for each of the alleles. Five sire lines (two Merino x Merino, two Merino x Coopworth, one Dorset Down x Coopworth) provided phenotypic and genotypic data used to ascertain the effects of allelic variation at the ADRB3 locus on birth weight, weaning weight, growth rate (up until weaning), carcass composition at 63 days post-weaning and cold survival. Statistical analyses within each half-sib family showed that in some sire lines (S13, S15, and S17) the inheritance of a particular allele was associated with increased birth weights and/or increased growth rates up until weaning. The inheritance of a particular sire allele was associated with fatter carcasses in sire line S16. Chi-squared analysis revealed the association of the E allele with cold survival and the D allele with cold-related mortality in sire line S14. Such associations support the hypothesis that ADRB3s are involved in energy homeostasis. With more research, the variation detected at the ADRB3 locus may assist in the genetic selection for desirable animal production traits. PMID- 12580783 TI - Genetic diversity and relationships of indigenous Kenyan camel (Camelus dromedarius) populations: implications for their classification. AB - The genetic diversity and relationships amongst the dromedary (Camelus dromedarius) populations are poorly documented. Four recognized Kenyan dromedary breeds (Somali, Turkana, Rendille, Gabbra) and dromedary from Pakistan and the Arabian Peninsula (Saudi Arabia, United Arab Emirates) were studied using 14 microsatellite loci. Phylogenetic analysis showed that Kenyan dromedaries are distinct from Arabian and Pakistani populations. Expected heterozygosity and allelic diversity values indicate that Kenyan dromedaries are less diverse than non-Kenyan populations. With the exception of the Somali population, the Kenyan dromedaries are poorly differentiated (average FST=0.009), with only one to two loci separating the Gabbra, Rendille and Turkana populations studied (P < 0.05). Individual assignments were performed using the maximum likelihood method. A correct breed assignment of only 39-48% was observed for the Kenyan dromedaries, using an allocation stringency of a log of the odds ratio >2. Our results do not support the present classification of the indigenous Kenyan dromedary into four distinct breeds based on socio-geographical criteria. Instead, our results point to just two separate genetic entities, the Somali and a group including the Gabbra, Rendille and Turkana populations. PMID- 12580784 TI - Genetic variation in caribou and reindeer (Rangifer tarandus). AB - Genetic variation at seven microsatellite DNA loci was quantified in 19 herds of wild caribou and domestic reindeer (Rangifer tarandus) from North America, Scandinavia and Russia. There is an average of 2.0-6.6 alleles per locus and observed individual heterozygosity of 0.33-0.50 in most herds. A herd on Svalbard Island, Scandinavia, is an exception, with relatively few alleles and low heterozygosity. The Central Arctic, Western Arctic and Porcupine River caribou herds in Alaska have similar allele frequencies and comprise one breeding population. Domestic reindeer in Alaska originated from transplants from Siberia, Russia, more than 100 years ago. Reindeer in Alaska and Siberia have different allele frequencies at several loci, but a relatively low level of genetic differentiation. Wild caribou and domestic reindeer in Alaska have significantly different allele frequencies at the seven loci, indicating that gene flow between reindeer and caribou in Alaska has been limited. PMID- 12580786 TI - Analysis of the 5' region of the canine PAX3 gene and exclusion as a candidate for Dalmatian deafness. AB - The causative mutation in a gene related to hearing loss in Dalmatians has been elusive. Because of its role in melanocyte migration and differentiation as integral component of the inner ear, we hypothesized that the canine PAX3 (paired box homeotic gene 3) gene could be a candidate for Dalmatian deafness. Therefore, we isolated the canine PAX3 gene and searched for causative mutations within the coding region of important regulatory domains of PAX3. However, no mutations were identified when comparing the DNA sequences of healthy and affected dogs. These results were confirmed by a two-point linkage analysis in 203 Dalmatians transmitting deafness. Our data clearly show that the canine PAX3 gene can be excluded as candidate for Dalmatian deafness. PMID- 12580785 TI - Comparative mapping of a region on chromosome 10 containing QTL for reproduction in swine. AB - Several quantitative trait loci (QTL) for important reproductive traits (age of puberty, ovulation rate, nipple number and plasma FSH) have been identified on the long arm of porcine chromosome 10. Bi-directional chromosome painting has shown that this region is homologous to human chromosome 10p. Because few microsatellite or type I markers have been placed on SSC10, we wanted to increase the density of known ESTs mapped in this region of the porcine genome. Genes were chosen for their position on human chromosome 10, sequence availability from the TIGR pig gene indices, and their potential as a candidate gene. The PCR primers were designed to amplify across introns or 3'-UTR to maximize single nucleotide polymorphism (SNP) discovery. Parents of the mapping population (one sire and seven dams) were amplified and sequenced to find informative markers. The SNPs were genotyped using primer extension and mass spectrometry. These amplification products were also used to probe a BAC library (RPCI-44, Roswell Park Cancer Institute) for positive clones and screened for microsatellites. Six genes from human chromosome 10p (AKR1C2, PRKCQ, ITIH2, ATP5C1, PIP5K2A and GAD2) were mapped in the MARC swine mapping population. Gene order was conserved within these markers from centromere to telomere of porcine chromosome 10q, as compared with human chromosome 10p. Four of these genes (PIP5K2A, ITIH2, GAD2 and AKR1C2), which map under QTL, are potential candidate genes. Identification of porcine homologues near important QTL and development of a comparative map for this chromosome will allow further fine- mapping and positional cloning of candidate genes affecting reproductive traits. PMID- 12580787 TI - X/XY/XYY mosaicism as a cause of subfertility in boars: a single case study. AB - Sex chromosome abnormalities are common in mammals and humans and are often associated with subfertility. In this study a boar with normal sperm parameters was indicated to have reduced prolificacy from figures obtained for return rate, farrowing rate and total number of piglets born. G-banded cytogenetic analysis of peripheral blood identified an abnormal mosaic sex chromosome constitution 39,XYY[74]/38,XY[23]/37,X[3]. Cytogenetic analysis of fibroblasts confirmed this mosaic karyotype with similar percentages of cell lines observed 39,XYY[76]/38,XY[19]/37,X[5]. External genitalia revealed a poorly developed scrotum with the right testicle being smaller than the left. To the best of our knowledge this is the first time that this chromosome constitution has been reported in the pig. It is of particular interest that this karyotype is associated with reduced boar fertility, which could lead to potential economic losses if such a boar were selected for breeding purposes. PMID- 12580788 TI - Genetic relationship between Mongolian and Norwegian horses? AB - Human populations of Central Asian origin have contributed genetic material to northern European populations. It is likely that migrating humans carried livestock to ensure food and ease transportation. Thus, eastern genes could also have dispersed to northern European livestock populations. Using microsatellite data, we here report that the essentially different genetic distances DA and (deltamu)2 and their corresponding phylogenetic trees show close associations between the Mongolian native horse and northern European horse breeds. The genetic distances between the northern European breeds and Standardbred/Thoroughbred, representing a southern-derived source of horses, were notably larger. We suggest that contribution of genetic material from eastern horses to northern European populations is likely to have occurred. PMID- 12580789 TI - Cloning, characterization and chromosomal localization of the Sus scrofa SLC31A1 gene. AB - Copper is an essential element necessary for normal function of numerous enzymes in all living organisms. Uptake of copper into the cell is thought to occur through the membrane protein, SLC31A1 (CTR1), which has been described in a variety of species including yeast, human and mouse. In this study, we present cloning, gene structure, chromosomal localization and expression pattern of the Sus scrofa SLC31A1 gene, which encodes a 189 amino acid protein. The (SSC) SLC31A1 gene is organized in four exons and spans an approximately 2.3 kb genomic region. We have localized the gene to chromosome 1q28-q2.13 using a somatic cell hybrid panel. This region shows conservation of synteny with human chromosome 9, where the human SLC31A1 (CTR1) gene has been localized. Expression studies suggest that SLC31A1 mRNA is transcribed in all tissues examined. PMID- 12580790 TI - Physical mapping of the brain and ovarian aromatase genes in the Nile Tilapia, Oreochromis niloticus, by fluorescence in situ hybridization. AB - Cytochrome P450-aromatase enzyme (CYP19), which catalyses the conversion of androgens to oestrogens, is critical in ovarian differentiation and hence in the sex differentiation pathways of non-mammalian vertebrates. As in other fish species, distinct ovarian and brain aromatase genes have been identified in the Nile Tilapia, Oreochromis niloticus. Here we demonstrate by in situ hybridization that the two aromatase genes of this species are present on different chromosomes and that neither are located on the sex chromosomes. Hence, the aromatase genes are not the primary sex determination genes in O. niloticus. PMID- 12580791 TI - Characterization and linkage map assignments for 61 new horse microsatellite loci (AHT49-109). PMID- 12580792 TI - The canine HMGA1 gene maps to CFA 23. PMID- 12580793 TI - Contribution of a new set of canine microsatellites to the knowledge of the canine genetic map. PMID- 12580794 TI - Eighty-three previously unreported equine microsatellite loci. PMID- 12580795 TI - KITLG maps to canine chromosome 15 and is excluded as a candidate gene for merle in dogs. PMID- 12580796 TI - Physical mapping of the rod cGMP-phosphodiesterase -subunit (PDE6G) gene to pig chromosome 12. PMID- 12580799 TI - Natural killer cells in the airways: Do they matter? PMID- 12580800 TI - Atopic eczema/dermatitis syndrome - a genetically complex disease. New advances in discovering the genetic contribution. PMID- 12580801 TI - Dustborne and airborne fungal propagules represent a different spectrum of fungi with differing relations to home characteristics. AB - BACKGROUND: Exposure to fungi is often assessed by culturing floor dust or air samples. Our objective was to evaluate the relationships between dustborne and airborne fungi and to identify factors that modify these relationships. METHODS: From November 1994 to September 1996 sequential duplicate 45-l air samples were collected in bedrooms of 496 homes in the Boston area, using a Burkard culture plate sampler. After air sampling, bedroom floors were sampled with a vacuum cleaner that was modified to collect dust in a cellulose extraction thimble. Dust was sieved, and the fine dust was dilution-plated onto DG-18 media. RESULTS: Concentrations of total culturable fungi per gram of bedroom-floor dust were correlated weakly, but significantly, with those of indoor air (r = 0.13, P < 0.05). Concentrations of some individual taxa in the dust and indoor air were also weakly associated. Adjusting for the concentrations of fungi in outdoor air, dustborne fungal concentrations were positively associated with those in indoor air for the taxa Cladosporium and Penicillium, but not for total fungi. The indoor air fungal levels were often predicted by different covariates to those predicting fungal levels in dust. The type of housing (house or apartment) and the presence of carpeting were often predictive factors for dust fungi. In contrast, outdoor fungal levels were often predictive of the indoor air fungal levels. CONCLUSIONS: Because our data do not indicate a strong overall relationship between culturable fungi in dust and indoor air, the results from these two methods (dust and air sampling) likely represent different types of potential fungal exposures to residents. It may be essential to collect both air and dust samples, as well as information on housing characteristics, as indicators for fungal exposure. PMID- 12580802 TI - Pets in the home and the development of pet allergy in adulthood. The Copenhagen Allergy Study. AB - BACKGROUND: The aim of the study was to investigate the association between exposure to cat and dog in the home and the development (incidence) of IgE sensitization to cat and dog. METHODS: Participants in a population-based study of 15-69-year-olds in 1990 were invited to a follow-up in 1998. Serum IgE antibodies against common inhalant allergens was assessed in 734 subjects (participation rate 69.0%) on two occasions 8 years apart. Information about current or previous keeping of cats and dogs in the home was obtained in a questionnaire at baseline. RESULTS: A cat in the home currently was significantly associated with the development of IgE sentisization to cat (adjusted odds ratio 8.4, 95% confidence interval (CI) 1.7-42.7). Moreover, an atopic predisposition in terms of IgE sensitization to allergens other than cat at baseline was an independent risk factor for the development of IgE sensitization to cat. A dog in the home was not significantly associated with the development of IgE sensitization to dog. CONCLUSIONS: In this adult population, exposure to a cat in the home increased the risk of developing IgE sensitization to cat. More prospective data are needed on this issue. PMID- 12580803 TI - Medical Indoor Environment Counselor (MIEC): role in compliance with advice on mite allergen avoidance and on mite allergen exposure. AB - BACKGROUND: In order to improve patient compliance in allergen avoidance, a new occupational activity was created: Medical Indoor Environment Counselor (MIEC). The aim of this study was to assess the impact of an MIEC on compliance with advice for mite allergen reduction in patients sensitized and exposed to mite allergens, and on mite allergen levels. METHODS: The study included 378 patients from four centers (Marseilles, Montpellier, Paris, Strasbourg) in a randomized prospective study. Patients in group A received advice from doctors only, and those in group B from doctors and MIEC. Mite allergen levels were measured on mattresses, mattress bases, and floors. Compliance and mite allergen levels were assessed 5 months later. RESULTS: Compliance for changing the mattress bases, removing or treating the carpets, washing duvets, pillows and stuffed animals were significantly higher in group B than in group A. A significant decrease in mite allergen was obtained for group B in mattress bases (83.8-22.9 micro g/g) and in carpets (15.8-6.3 micro g/g), but not in group A. CONCLUSION: Our results suggest that the home visit by the MIEC 1) increased the compliance to mite reduction methods advised; 2) induced a significant difference in mite reduction levels on mattress bases and on carpets; 3) avoided nonestablished avoidance advice. PMID- 12580804 TI - Isolation and characterization of Tha p 1, a major allergen from the pine processionary caterpillar Thaumetopoea pityocampa. AB - BACKGROUND: Pine processionary caterpillars induce dermatitis by a toxic irritative mechanism. The existence of true allergic reactions to allergens from these caterpillars has been recently demonstrated by positive immediate skin prick tests and specific IgE determination by immunoblotting using crude larval extracts. The aim of this work was to purify allergens from the crude larval extract in order to characterize IgE-binding proteins from these caterpillars. METHODS: Allergens were separated by ethanol gradient fractionation and reversed phase HPLC. The N-terminal amino acid sequence of a selected allergen was obtained after SDS-PAGE and transfer. The clinical relevance of this allergen was measured using sera from patients allergic to caterpillar. RESULTS: An allergen with a molecular weight close to 15 kDa was purified. It was recognized by 9 out of 11 allergic patients (82%). Its N-terminal amino acid sequence had no homologies to any other protein already described in data bases. For this reason, no information about its biological function could be obtained. CONCLUSIONS: This 15-kDa IgE-binding protein is a major caterpillar allergen and shows no homologies to other insect allergens already described. PMID- 12580805 TI - A comparison of different allergometric tests, skin prick test, Pharmacia UniCAP and ADVIA Centaur, for diagnosis of allergic diseases in children. AB - BACKGROUND: The diagnosis of allergic disease is performed by skin prick tests (SPT) or through the demonstration of specific IgE in a blood sample via an in vitro test. The measurement of IgE concentration against allergens provides critical information in clinical allergy. Standardized and reproducible methods contribute to the quality of diagnosis and treatment of allergic disease. METHODS: In this study we evaluated the performance of a new specific IgE method, developed by ALK-Abello for Bayer Diagnostics to run on their ADVIA Centaur immunoassay system. One hundred and fifty-one children with allergic diseases (both food and inhalant allergies) were tested for specific IgE (sIgE) via SPT and in vitro tests (UniCAP system, Pharmacia, and ADVIA Centaur immunoassay system, Bayer Diagnostics) and the test results were correlated with the clinical data. RESULTS: Statistical analysis revealed no significant difference between the two in vitro tests compared with clinical history. The sensitivities and specificities are similar, but the UniCAP system method has higher sensitivity. In the children with cow's milk allergy, the UniCAP system has sensitivity of 91% and specificity of 70%; the ADVIA Centaur immunoassay has sensitivity of 82% and specificity of 74%. In hen's egg allergy, UniCAP system has 94% sensitivity and 64% specificity, and the ADVIA Centaur system has 88% sensitivity and 52% specificity. In inhalant allergies, the two methods show statistically similar performances for both grass pollen allergies (UniCAP sensitivity 100%, specificity 73%; ADVIA Centaur sensitivity 95%, specificity 79%) and in the dust mites allergies (UniCAP sensitivity 91%, specificity 62%; ADVIA Centaur sensitivity 86%, specificity 64%). In cat allergies, the systems showed equivalent results (UniCAP sensitivity 100%, specificity 71%; ADVIA Centaur sensitivity 100%, specificity 70%). Using the UniCAP system, the geometric mean of sIgE values in children with clinical allergy is significantly higher than in sensitized ones. The ADVIA Centaur system shows a similar trend with the exclusion of cow's milk and Dermatophagoides farinae allergens. With this last method the mean value of sIgE is higher in sensitized than in symptomatic children. CONCLUSION: The new ADVIA Centaur method compares favorably with the results obtained on the UniCAP system. If other studies continue to confirm this data, then the advantages are numerous: the use of only a small quantity of serum (25 micro l per allergen), rapid turnaround time, minimal hands-on time, and no interference from IgG. PMID- 12580806 TI - Relationship of skin-prick reactivity to aeroallergens and hyperresponsiveness to challenges with methacholine and adenosine monophosphate. AB - BACKGROUND: Clarification of the relationship between atopy and bronchial hyperresponsiveness (BHR), both key features of asthma, is critical to our understanding of the disease. We therefore investigated the putative relationship between skin-prick reactivity to aeroallergens and BHR to direct and indirect stimuli. METHODS: We performed a retrospective analysis of data from 332 patients presenting with a diagnosis of asthma. Patients were characterized by skin prick tests (SPT), spirometry and bronchial challenge with methacholine and adenosine monophosphate (AMP). RESULTS: For patients who had BHR to methacholine but not AMP, the presence of atopy was associated with a lower PD20 (the provocative dose of methacholine producing a fall in FEV1 of 20%), amounting to a geometric mean (95% confidence interval (CI)) of 2.3-fold (1.4-4.0) difference. Furthermore, the number of skin-prick positive (SPP) responses was related to methacholine reactivity: 0-1 SPP, PD20 = 69.9 micro g; 2-4 SPP, PD20 = 47.8 micro g; 5-8 SPP, PD20 = 35.6 micro g. There was a 2.0- fold (1.1-3.6) difference between the groups with a low (0-1 SPP) and high (5-8 SPP) degree of skin-prick reactivity. A similar pattern was seen when data were analyzed including only perennial allergens. Spirometry was unrelated to the degree of skin-prick reactivity. DISCUSSION: These results suggest that skin-prick reactivity to aeroallergens is associated with BHR to methacholine. PMID- 12580807 TI - Skin test reactivity to natural and recombinant Blomia and Dermatophagoides spp. allergens among mite allergic patients in the UK. AB - BACKGROUND: Many asthmatics in tropical and subtropical areas have positive skin prick tests to both Dermatophagoides spp. and to the mite Blomia tropicalis. This may be due to recognition by IgE of cross-reactive allergens between the different mite species or because of sensitization to species-specific allergens. A 14-kDa Blomia tropicalis allergen, Blo t 5, has been cloned and shows 40% sequence homology with Der p 5. The aim of this study was to investigate reactivity to B. tropicalis in patients known to be sensitized to D. pteronyssinus and to assess allergenic activity and cross-reactivity of recombinant (r) Group 5 allergens amongst these patients, who live in the UK and who are not exposed to B. tropicalis in their homes. METHODS: Patients (n = 19) with asthma and/or rhinitis were selected based on clinical history and a positive skin prick test to D. pteronyssinus extract and were compared with non allergic skin test negative controls (n = 10). IgE antibody responses to Blomia tropicalis, Dermatophagoides pteronyssinus, rDer p 5 and rBlo t 5 were compared by quantitative intradermal skin testing using serial 10-fold dilutions of each allergen. End point titre was the highest dilution giving an 8 x 8 mm wheal at 15 min. IgE antibodies to Blomia tropicalis, Dermatophagoides pteronyssinus, rDer p 5 and rBlo t 5 were measured using RAST, CAP and RIA, respectively. RESULTS: All 19 patients had positive skin tests to D. pteronyssinus at concentrations of 0.001 to 1 AU/ml and 10 were skin test positive to rDer p 5 at concentrations of 10-4 to 5 micro g/ml. Positive intradermal tests to Blomia tropicalis were seen in 12/19 patients at concentrations of 0.002 to 2 micro g/ml. However none of the patients had positive skin tests to rBlo t 5. Non-allergic controls were all skin test negative at the highest concentration of each allergen tested. All subjects had quantifiable specific IgE to D. pteronyssinus, but only two had IgE to B. tropicalis. IgE to Der p 5 was found in six patients, but no patients had IgE to Blo t 5. CONCLUSIONS: This study of patients naturally exposed to D. pteronyssinus but not to Blomia tropicalis, provides evidence for IgE mediated cross-reactivity between allergens produced by both mite species. The results suggest that the Group 5 allergens of D. pteronyssinus and B. tropicalis are species-specific. PMID- 12580808 TI - Latex hypersensitivity: relationship with positive prick test and patch test responses among hairdressers. AB - BACKGROUND: Natural rubber latex is a frequent cause of IgE-mediated allergy in hairdressers; but a non-IgE-mediated allergy to latex proteins can also occur. Sixty-one hairdressers, reporting latex glove-related symptoms, were enrolled in the study. METHODS: They were evaluated for latex allergy by assessment of serum specific IgE and skin prick tests. Patients were patch tested to a rubber additive series and to latex. Proportions were compared by one-tailed Fisher's exact test; mean values of continuous variables were compared by Student's t test. RESULTS: A diagnosis of IgE-mediated latex allergy was made in 11 (18%) hairdressers; patch tests with rubber-related antigens were positive in 24 (39.3%) subjects. The most positive patch tests were those with para phenylenediamine (33%). Two hairdressers (3.3%) showed delayed reactions to the latex sample. Hairdressers with IgE-mediated latex allergy were older and presented a personal history of atopy and local hives after exposure to latex; they were more likely to have a positive response to other allergens on prick or patch testing. CONCLUSIONS: Our study demonstrates a certain number of false positive and false-negative results when reported symptoms of possible latex sensitivity were compared to prick and CAP tests to latex, indicating the problems of relying solely on self-report. PMID- 12580809 TI - In vitro kinetics of allergen- and microbe-induced IL-4 and IFN-gamma mRNA expression in PBMC of pollen-allergic patients. AB - BACKGROUND: According to a hypothesis allergens induce Th2 responses in allergic patients, and microbes induce Th1 responses. We studied the kinetics of in vitro allergen-, tuberculin (PPD)- and tetanus toxin (TT)-induced IFN-gamma and IL-4 mRNA expression in peripheral blood mononuclear cell (PBMC) cultures of pollen allergic patients and healthy controls. METHODS: PBMC of 10 birch or timothy pollen-allergic patients and of 13 healthy controls were stimulated in vitro with allergen (birch or timothy), PPD or TT. Pellets and supernatants were collected at 24, 48, 72 and 96 h after stimulation. IFN-gamma and IL-4 production was measured by enzyme linked immunosorbent assay and mRNA expression using RT-PCR and time-resolved fluorometry. RESULTS: Allergen induced IFN-gamma production and mRNA expression in PBMC more in allergic patients than in healthy controls. Also allergen induced IL-4 mRNA expression more in allergic patients than in healthy controls. PPD induced IFN-gamma mRNA expression both in allergic patients and healthy controls, whereas IFN-gamma production was induced only in healthy controls and IL-4 was not induced at all. TT induced IFN-gamma mRNA expression in both groups, IFN-gamma production in allergic patients, and IL-4 mRNA expression in both allergic patients and healthy controls. CONCLUSIONS: In vitro stimulation with allergen induced both IFN-gamma and IL-4 mRNA expression of PBMC in allergic patients. These observations challenge the clearcut division of microbe-specific Th1 and allergen-specific Th2 responses in peripheral blood. PMID- 12580810 TI - Decreased intercellular adhesion molecule-1 (CD54) and L-selectin (CD62L) expression on peripheral blood natural killer cells in asthmatic children with acute exacerbation. AB - BACKGROUND: The capacity of inflammatory cells to adhere involves an array of adhesion molecules, and is critical to the inflammatory responses seen in childhood asthma. We aimed to determine the changes of intercellular adhesion molecule-1 (ICAM-1) and L-selectin expressed on peripheral blood (PB) T lymphocytes and natural killer (NK) cells in asthmatic children with acute exacerbation and after prednisolone therapy. METHODS: Flow cytometric analysis was performed to determine the expression of ICAM-1 (CD54) and L-selectin (CD62L) on T (CD3+) cells and NK (CD3-/CD56+) cells of PB from children with allergic asthma with acute exacerbation and in a stable condition after prednisolone therapy. Atopic subjects without asthma and age-matched controls were also included for comparison. RESULTS: Percentages of PB non-CD3, CD56+ NK cells, but not CD3+ T cells, increased in asthmatic children with acute exacerbation, compared to those assessed in a stable condition after a course of prednisolone. However, significant decrease of ICAM-1 (P = 0.01) and L-selectin (P = 0.01) expression on PB NK cells, but not on T cells, were found in children with acute asthma compared to those in a stable condition. NK cells in children with acute asthma showed minimal expression of CD69 and CD25. CONCLUSIONS: Results suggests that either NK cells expressing ICAM-1 and L-selectin selectively migrated into inflamed lung tissues, or subsets of NK cells not expressing ICAM-1/L-selectin were expanded during acute exacerbation of childhood asthma. PMID- 12580811 TI - HLA-DR-dependent increased mannan-induced lymphoproliferative response in atopic eczema dermatitis syndrome. AB - BACKGROUND: In yeast-sensitive atopic eczema dermatitis syndrome (AEDS), yeast mannan induces highly elevated specific IgE levels and lymphoproliferative responses. In healthy individuals the involvement of both human leukocyte antigen (HLA)-dependent T-cell activation and non-HLA-dependent activation, e.g. by crosslinking of the cell surface mannose receptors, has been suggested. In the present study the HLA dependence and the role of crosslinking in the lymphoproliferative response to mannan in AEDS has been analyzed. METHODS: Twenty patients with AEDS and 12 controls with no history of allergic diseases were included in the study. Mannan from Candida albicans was prepared according to the Cetavlon method. Following isolation using Ficoll-Hypaque, peripheral blood mononuclear cells (PBMC) were incubated with the mannan preparation in the absence and presence of different concentrations of neutralizing anti-HLA antibodies and alpha-methylmannoside for 6 days and proliferative responses were measured by 3H-thymidine incorporation and scintilloradiography. RESULTS: In AEDS patients with elevated mannan-specific serum IgE, the C. albicans mannan induced lymphoproliferation. Mannan-induced lymphoproliferative responses could be inhibited, dose-dependently, by neutralizing anti-HLA-DR, but not anti-HLA-DQ antibodies in AEDS patients and healthy controls. The addition of alpha methylmannoside, that blocks binding to mannose receptors, inhibited lymphoproliferative responses in a dose-dependent way by 50% only in healthy controls, but not in AEDS patients. Levels of inhibition of the proliferation by alpha-methylmannoside correlated inversely with the yeast- and mannan-specific IgE levels. CONCLUSIONS: These results show that in healthy subjects yeast mannan activates lymphocytes both in an HLA-DR-dependent manner and as a result of direct crosslinking of the cell surface. However, in AEDS the elevated lymphoproliferative response is HLA-DR-dependent, although only a slight proportion of this response results from direct crosslinking. PMID- 12580812 TI - The upper and lower airway responses to nasal challenge with house-dust mite Blomia tropicalis. AB - BACKGROUND: The house dust mite Blomia tropicalis (B. tropicalis) was found to be the most prevalent domestic mite in Singapore. However, its pathogenicity in allergic airway diseases remains to be investigated. METHODS: Twenty adults with persistent allergic rhinitis (PAR) were studied. Five had a history of asthma, and all were asymptomatic except one who was under treatment with low-dose inhaled corticosteroid. Nasal challenge was carried out by nasal spray with phosphate-buffered saline (PBS) and with increasing concentrations of crude B. tropicalis extracts (0.6, 6.0 and 60 micro g/ml) at 15 min intervals. Subjective symptom scores and absolute number of sneezes were recorded together with objective measurements of spirometry (forced expiratory volume in 1 s, FEV1) and acoustic rhinomanometry (volume of the nasal cavity). These were performed at baseline, 5 min after each incremental challenge, and 30 min, 1 h, 3 h, 5 h and 7 h after the last challenge. Meanwhile, concentrations of mediators in nasal secretions (tryptase, leukotriene C4 (LTC4) and eosinophil cationic protein (ECP)) were measured in nasal aspirate samples at similar time intervals. An identical (control) challenge procedure with PBS alone was repeated in seven patients after a washout period of at least 2 weeks. RESULTS: Significant increases in the subjective and objective nasal symptoms, together with a significant increase of tryptase and LTC4 concentrations in nasal secretion, were found 5 min after each challenge with B. tropicalis, but not with PBS. There was no definitive pattern of the late-phase nasal response in either subjective symptoms or objective measurements. Three patients (3/5) with a history of asthma showed a fall in FEV1 readings (33%, 22% and 11% from baseline, respectively) at 7 h post challenge with concomitant mild wheezing in the night. CONCLUSIONS: Our study demonstrates direct evidence of allergic nasal response to B. tropicalis in sensitized adults. It shows that nasal provocation may also provoke concomitant asthmatic symptoms during the late-phase reaction, especially in people with a history of asthma. PMID- 12580813 TI - House-dust mite allergens in Italy and Spain. PMID- 12580814 TI - Delayed skin reactions to metamizol. PMID- 12580815 TI - Occupational asthma to grain pests. PMID- 12580816 TI - Vascular rings can give cough. PMID- 12580817 TI - Skin reactions to pine processionary caterpillar. PMID- 12580818 TI - Gentamicin-induced anaphylaxis. PMID- 12580819 TI - Severe hypoproteinemia in infant with AD. PMID- 12580821 TI - Inequality and inequity in use of mental health services. PMID- 12580822 TI - Addiction, stigma and movies. AB - OBJECTIVE: To identify common character stereotypes of alcohol and other drug users as portrayed in motion pictures. METHOD: A selective review of a number of movies prominently portraying alcohol and other drug use and misuse. RESULTS: The great majority of popular films portray alcohol and drug use whether as a routinized background, routinized foreground or exceptional foreground. Four main stereotypes of alcohol and other drug users appear to be prevalent - the tragic hero, the demonized user, the rebellious free spirit and the comedic user. A number of movies are selected which portray alcohol and other drug use as a prominent theme. CONCLUSION: Movies, as a medium for mass communication, have a powerful influence on the public and perpetuate popular mythologies regarding alcohol and other drug use. PMID- 12580823 TI - Socio-economic differences in psychiatric in-patient care. AB - OBJECTIVE: We seek to investigate socio-economic differences in psychiatric in patient care regarding admission, treatment and outcome. METHOD: This study is undertaken on a comprehensive and exhaustive psychiatric case register of all psychiatric in-patient care carried out in Belgium in 1997 and 1998 (n=144 754). RESULTS: Lower socio-economic groups were more likely to be compulsorily admitted, to be cared for in a non-teaching or psychiatric hospital, to be admitted in a hospital with unexpectedly long average length of stay and to be admitted to a ward with a more severe case-mix. They were less likely to receive antidepressants and psychotherapies. The improvements in functioning and in symptoms were also less favourable for these groups. The lowest group had a higher risk of dying in the hospital. CONCLUSION: Psychiatric in-patient care is associated with moderate socio-economic differences in access, treatment and outcome. Further research is needed to clarify the causes of such disparities. PMID- 12580824 TI - Does educational background explain inequalities in care service use for mental health problems in the Dutch general population? AB - OBJECTIVE: To investigate whether (1) education predicts the use of care services for mental health problems, independently of mental disorder and functional impairment and (2) education modifies the association between mental disorder and service use. METHOD: Predictors of service use were recorded at baseline, and service use itself at 12-month follow-up, in a representative sample (N=7076) of the Dutch population, using the Composite International Diagnostic Interview. RESULTS: People with more education were less likely to use primary care but more likely to use mental health care. The effects on both types of care remained significant after adjustment for mental disorder and functional impairment. Lower education tended to strengthen the association between mood or anxiety disorder and primary care use. CONCLUSION: Further research on inequalities in service use will benefit from additional explanatory analyses and from the inclusion of sociopsychological variables, like cost-benefit considerations in decisions to use services. PMID- 12580825 TI - What patient characteristics make clinicians recommend brief treatment? AB - OBJECTIVE: Assessing self-rated items that might have an impact on clinicians recommending brief treatment (BT) over unlimited or long-term treatment (ULT). METHOD: On the basis of patient self-report data we compared patients referred by clinicians to BT (n=71) with those referred to ULT (n=145). RESULTS: The final multiple logistic regression model indicates that the chance of being allocated to BT increases with: more satisfaction with support, higher self-esteem, primary education or less, and high desire for support as an intervention. With regard to desire to confess in treatment, low and high scores make the chance of being allocated to BT lower. This is also the case for daily hassles. Finally, some specific target complaints, in particular anxiety, lower the chance of being allocated to BT. CONCLUSION: Using data about patient's complaints and symptoms, stress and support, personality and coping, and request for type of intervention, we built a regression-model that classified 80% of the patients correctly with regard to allocation to BT or ULT. PMID- 12580826 TI - Gender and depression: a study of severity and symptomatology of depressive disorders (ICD-10) in general practice. AB - OBJECTIVE: Gender differences in severity and symptomatology of depression in a large and representative sample of depressive patients from general practice were examined. METHOD: During a 2-year study period, 31 Danish general practitioners consecutively assessed patients, aged >18, for depression. A total of 1033 patients (692 female, 341 male) fulfilled ICD-10 criteria for depressive disorders. Gender differences were examined with regard to severity and symptomatology of depression. RESULTS: The severity of depression was similar for men and women with the following distribution: 22% mild, 56% moderate and 23% severe depressive episodes. The mean number of symptoms presented was 6.6 of 10 for both genders. There were no gender differences in prevalence of single depressive symptoms. CONCLUSION: No gender differences in the severity or symptomatology of depression were found in a highly representative sample of patients with depressive disorders. PMID- 12580827 TI - Affective disorder and 'psychopathy' in a sample of younger male delinquents. AB - OBJECTIVE: The authors' goal was to determine the prevalence of affective disorder, a history of traumatic events and the prevalence of post-traumatic stress disorder (PTSD) diagnosis in their relationship to 'psychopathy' in a sample of younger male delinquents admitted to a correctional institution. METHOD: As part of routine mental health screening, the SCID-1 and Psychopathy Checklist (PCL-R) were administered to 102 inmates aged 17-27 years. RESULTS: Affective disorder was found in 28% and 29% had a history of suicidal attempts. The PCL-R-score was significantly correlated with the number of prior threatening events, but PTSD could not be diagnosed in the 'psychopathic' group. The prevalence of affective disorder was significantly higher in the 'non psychopathic' group. CONCLUSION: The inverse relationship between threatening events and the PTSD diagnosis seems to indicate other coping modalities than those found in 'non-psychopaths'. That finding is discussed against the background of psychophysiological data and psychosocial learning models. PMID- 12580828 TI - Stressful life events and affective illness. AB - OBJECTIVE: To investigate the association between the occurrence of affective illnesses and the number/type of experienced negative stressful life events in a twin material. A case-control study with an unrelated twin as control to the case and a co-twin control study were both undertaken with the same material. METHOD: Postal questionnaire responses were used for confirming diagnosis and to inventory stressful life events. Risk ratios (RR) for groups of events were calculated using conditional logistic regression. RESULTS: A dose-response relationship was observed for the association between stressful life events and affective illness. The RRs for specific groups of exposures were higher in the co twin control study and higher for dizygotic twin pairs than for monozygotic twin pairs. CONCLUSION: Individuals with a history of affective illness select themselves into high-risk environments, in part due to their genetic propensity to the disease. Thus, the association represents a classic genotype-environment correlation. PMID- 12580829 TI - Evaluation of the psychometric characteristics of the Spanish version of the Hospital Anxiety and Depression Scale. AB - OBJECTIVE: To assess the psychometric properties of the Spanish version of the Hospital Anxiety and Depression Scale(HADS). METHOD: We administered HADS to 685 participants (256 controls and 429 patients with five different diagnoses). The reliability of the instrument was assessed by a test-retest study. Construct validity studies were carried out through item-subscale correlation and factor analysis for the whole group and by each of the five different diagnoses. Three instruments were used as external criteria to assess concurrent validity. RESULTS: HADS test-retest reliability presented correlation coefficients above 0.85. The internal consistency was high, with a Cronbach's alpha of 0.86 (anxiety) and 0.86 (depression). Factor analysis showed a clear two-factor structure for all groups. The results showed high concurrent validity with the Beck Depression Inventory and State-Trait Anxiety Inventory and with the mental domains of the Short-Form Health Survey. CONCLUSION: The Spanish version of the HADS demonstrated good reliability and validity when used in medical patients. PMID- 12580830 TI - The European Schizophrenia Outpatient Health Outcomes (SOHO) study: rationale, methods and recruitment. AB - OBJECTIVE: The objective of the European Schizophrenia Outpatient Health Outcomes (SOHO) study is to understand the comparative costs and outcomes of antipsychotic drug treatment, with specific focus on olanzapine. The study will also provide a large database for research into the treatment and outcome of schizophrenia. The role of observational studies in the assessment of the effectiveness of antipsychotic agents is reviewed, and the rationale, design and recruitment issues surrounding the SOHO study are presented. METHOD: SOHO is a 3-year, prospective, observational study of the health outcomes associated with antipsychotic treatment in Europe. RESULTS: Over 10 000 patients have been recruited from 10 countries. Baseline evaluation included measures of clinical status, social functioning, quality of life, service use and pharmacological treatment. Patients will be followed for 3 years. CONCLUSION: The SOHO study will complement randomized controlled trial findings on the treatment of schizophrenia and will address relevant clinical and policy research questions. PMID- 12580831 TI - Alterations of rCBF and mitochondrial dysfunction in major depressive disorder: a case report. AB - OBJECTIVE: A mitochondrial disease might be considered when depressive disorder is associated with diabetes mellitus or other symptoms commonly found in mitochondrial disease. Scattered regional cerebral blood flow (rCBF) decreases and increases have been reported in depressive and mitochondrial disorders. A 61 year-old male patient with early adult onset of depressive disorder and a slowly developing multiorgan syndrome including diabetes mellitus was investigated. METHOD: 99mTc-HMPAO rCBF SPECT and muscle biopsy to assess mitochondrial functions were performed in the patient. RESULTS: Alterations of rCBF were found in the patient, with the most pronounced decreases in the left dorsolateral frontal and inferior parietal lobes, and the most pronounced increases in the bilateral superior parietal lobes. Muscle biopsy revealed myopathy and decrease of mitochondrial adenosine triphosphate production rates (MAPRs). CONCLUSION: The MAPRs decreases support the suspicion of mitochondrial dysfunction in the patient. A subgroup of depressed patients may have mitochondrial dysfunctions. PMID- 12580832 TI - An update on the controversies of tocolytic therapy for the prevention of preterm birth. AB - Preterm birth is the major cause of perinatal mortality and morbidity in the developed world. Where there are no contraindications to their use, tocolytics can improve neonatal survival rates by approximately 3% per day between 23 and 27 weeks gestation with a concomitant reduction in morbidity. The ultimate aim of tocolytic therapy is to prolong pregnancy until growth and maturation is complete, but even short-term delay may enable the administration of antepartum glucocorticoids to reduce hyaline membrane disease or to arrange transfer to a center with neonatal intensive care facilities. Both of these have been shown to reduce neonatal mortality and morbidity. Until recently, none of the currently used tocolytics, whether licensed or unlicensed, were developed specifically for the inhibition of preterm labor and consequently, they exhibit various potentially serious side-effects. As a result of the recent licensing of the oxytocin antagonist, atosiban, developed for the treatment of preterm labor and due to its high utero-specificity, obstetricians have experienced an advance in their options for the management of spontaneous preterm labor. PMID- 12580834 TI - Twin pregnancy outcome and chorionicity. AB - BACKGROUND: Based on research of relatively poor quality, it is generally believed that dichorionic twins have lower perinatal mortality than monochorionic twins. We assessed the relationship between the pattern of perinatal loss in twin pregnancy and chorionicity. METHODS: A cohort study of 238 consecutive sets of twin pregnancies registered in our antenatal service over a 2-year period (1996 98) had chorionicity determined by ultrasound at 12-16 weeks' gestation. Follow up included scanning at 20 weeks' gestation for anomaly, and at 2-3-weekly intervals from 24 weeks' gestation onwards for growth and well being. Hazard ratios were computed for comparing the risk of death according to chorionicity. The perinatal loss patterns were analyzed according to gestational age at demize and that at delivery using survival analysis. RESULTS: Overall, 17/238 (7.1%) twin pregnancies suffered mortality: 14/190 (7.3%) amongst the dichorionic and 3/48 (6.2%) amongst the monochorionic twins. The hazard ratio for mortality was 0.89 (95% confidence interval 0.27-2.97, p = 0.85), considering gestational age at demize as the outcome. For gestational age at delivery as the outcome, the hazard ratio for mortality was 0.93 (95% confidence interval 0.27-3.15, p = 0.91). Survival analysis showed that amongst the dichorionic twins the hazard of death continued to rise throughout gestation. In contrast, the hazard of death for the monochorionic twins rose gradually to a maximum at 28 weeks' gestation and was then constant. CONCLUSION: Chorionicity did not affect the overall fetal loss rate amongst the twin pregnancies in our setting. There were differences in patterns of loss according to chorionicity, which require further investigation. PMID- 12580833 TI - An evaluation of midwives' counseling of pregnant women in fear of childbirth. AB - BACKGROUND: The aim of this clinical evaluation was to study birth experience, post-traumatic stress symptoms, and satisfaction with care in new mothers who had consulted specially trained midwives because of a fear of childbirth during pregnancy. METHODS: Sixty-two women were eligible for the study, of whom 53 (85%) participated at 1-14 months postpartum. For comparison, a group of 53 women were matched for parity and mode of delivery. All 106 women completed two self-rating scales, the Wijma Delivery Experience Questionnaire (W-DEQ) and the Impact of Event Scale (IES), and answered several open questions about their opinion of the antenatal preparation given. RESULTS: Those women who had been treated for fear of childbirth reported a rather more frightening experience of delivery, and more frequent symptoms of post-traumatic stress related to delivery than did the women in the comparison group. Nevertheless, satisfaction with care was manifest in the study group. CONCLUSIONS: Women who seek help for fear of childbirth are a vulnerable group. Because the counseling received by the women in this study did not accord them the same positive experience of childbirth as the average parturient at the unit, more effective forms of treatment may be necessary. However, as most of the women were very satisfied with their care and with the outcome, one may assume that the care given had improved their situation to some degree. PMID- 12580835 TI - Elevated maternal mid-trimester chorionic gonadotropin > or =4 MoM is associated with fetal cerebral blood flow redistribution. AB - BACKGROUND: Elevated mid-trimester human chorionic gonadotropin (hCG) is associated with adverse maternal and perinatal outcome. The aims of the study were to evaluate the association between elevated hCG, fetal pathological arterial waveforms and maternal and perinatal complications. METHODS: Pulsatility indices (PI) of middle cerebral artery (MCA) and umbilical artery (UA) were determined prospectively in 121 consecutive patients with abnormal maternal serum hCG (> 2.5 MoM). Each patient had four US scans during pregnancy. Patients with known structural or chromosomal anomalies were excluded. RESULTS: Of 121 women with hCG > 2.5 MoM, 36/121(29.6%) had hCG between 2.5 and 3.0 MoM, 35/121(28.9%) had hCG between 3.0 and 3.5 MoM, 21/121(17.3%) had hCG of 3.5-4.0 MoM, 17/121(14.1%) had hCG levels between 4.0 and 4.5 MoM, and 12/121(9.9%) had hCG > 4.5 MoM. Middle cerebral artery PI was significantly lower in women with hCG > 4.0 MoM between 28 and 36 weeks' gestation, but not between 18 and 27 weeks' gestation. No differences of MCA PI were found when the cut-off point of hCG was 3.5. Women with hCG levels > 4.0 MoM had a significantly higher rate of preterm deliveries, cesarean sections, higher rate of Apgar scores < 7 and a significantly lower mean birth weight in comparison with women with hCG < 4.0 MoM. The prevalence of PIH and preeclampsia and perinatal death were found to be higher among patients with hCG levels > 4.0 MoM, although not significantly. No differences were found at hCG levels less than 4.0 MoM. CONCLUSIONS: In pregnancies with mid-trimester hCG > 4.0 MoM, redistribution of cerebral blood flow is expressed after 28 weeks' gestation. These pregnancies have higher rates of maternal and neonatal complications as compared to pregnancies with lower hCG levels. PMID- 12580836 TI - Cord blood thyroid-stimulating hormone level in twin pregnancy. AB - BACKGROUND: Cord blood thyroid-stimulating hormone level is elevated among neonates who undergo more perinatal stress. The aim of this study was to investigate the effect of twin pregnancy on the cord blood thyroid-stimulating hormone level. METHODS: The study group consisted of 24 910 singleton and 543 twin neonates over a 4-year period. The effect of twin pregnancy on the cord blood thyroid-stimulating level was evaluated. Linear regression analysis was used to control for the effect of mode of delivery, birth weight, and infant sex. RESULTS: The median cord blood thyroid-stimulating hormone levels (interquartile range) in the singletons and in the twins were 5.8mIU/l (4.2-8.6) and 5.6mIU/l (4.3-7.5), respectively. Linear regression analysis revealed no significant difference between singleton and twin cord blood thyroid-stimulating hormone levels (p = 0.23). Cord thyroid-stimulating hormone levels tended to be higher in second-born twins (p = 0.08) and monochorionic twins (p = 0.097) compared with singleton neonates. Twins with more than 20% weight discordance were associated with a significantly higher cord blood thyroid-stimulating hormone level (p = 0.035). CONCLUSION: Cord blood thyroid-stimulating hormone level is elevated in some subgroups of twins who are at higher risk of adverse perinatal outcomes. However, the overall effect of multiple pregnancy on the cord blood thyroid stimulating hormone level is small. Cord blood thyroid-stimulating hormone screening for congenital hypothyroidism is also valid in twin pregnancies. PMID- 12580837 TI - Relationship between umbilical cord morphology and nuchal cord entanglement. AB - OBJECTIVE: To determine whether tightening of nuchal cord entanglement is more likely to occur where normal cord coiling is absent and there is deficient Wharton's jelly. The study was carried out in the delivery suite of a university teaching hospital, using a prospective matched controlled-pairs survey. METHODS: Midwives identified index cases with nuchal cord entanglement at the time of delivery. The next delivery of equivalent gestation (+/- 1 week) without cord entanglement was taken as the control. Only singleton pregnancies with cephalic presentation undergoing spontaneous labor were studied. Placental insertion, nuchal entanglement (tight or loose), length, presence of knots, umbilical coiling index (UCI), and the amount of Wharton's jelly were recorded. RESULTS: Longer cords were more frequent amongst male infants and were predisposed towards entanglement. There were no significant differences in the amount of Wharton's jelly, or in the UCI between tight and loose nuchal cord groups. The only measurement that differed significantly was the total cord length, with shorter cords predisposed towards tightening. CONCLUSION: Whilst nuchal cord entanglement amongst male babies is frequently associated with long and straight umbilical cords, tightening is more likely to occur if the cord is of short or normal length. Neither low UCI nor low percentage of Wharton's jelly are predisposed towards tightening of cord entanglement. PMID- 12580838 TI - Trends in teenage fertility, abortion, and pregnancy rates in Iceland compared with other Nordic countries, 1976-99. AB - BACKGROUND: Iceland is often considered very similar to the other Nordic countries. The purpose of this study was to explore trends in teenage fertility, abortion, and pregnancy rates in Iceland, compare these trends with corresponding rates in Denmark, Finland, Norway, and Sweden during the period 1976-99, and to evaluate similarities and dissimilarities. METHODS: The study is based on data about fertility, abortion, and pregnancy rates obtained from the Icelandic and Nordic national population and abortion registers for the age group 15-19 years years. RESULTS: Teenage fertility and pregnancy rates in the five Nordic countries declined over the study period by 57-67% and 31-50%, respectively, and in Iceland they remained significantly higher than in the Nordic countries. In 1999 almost every other teenage pregnancy in Iceland (45.9/1000) resulted in a childbirth (24.4/1000). Regional fertility rates were highest in the countryside. While the abortion rate has been declining in the four Nordic countries by 20 41%, they have concurrently been rising in Iceland by 133% (9.4/1000 in 1976-80, 21.9/1000 in 1996-99) and are presently higher than in the other Nordic countries. Regionally, abortion rates in Iceland were highest in the Capital area. CONCLUSIONS: The teenage pregnancy rate is higher in Iceland than in the other Nordic countries. This may be explained by cultural norms in Iceland's society regarding childbearing, early initiation of sexual intercourse, more limited sex education, and less effective delivery and use of contraceptive methods. There is a need to promote sexual and reproductive health to young people in Iceland by combining diverse preventive approaches. PMID- 12580839 TI - The use of clomiphene citrate/human menopausal gonadotrophins in conjunction with GnRH antagonist in an IVF/ICSI program is not a cost effective protocol. AB - OBJECTIVE: To evaluate the cost effectiveness of a clomiphene citrate (CC)/human menopausal gonadotropin (hMG)/GnRH antagonist protocol versus a long-acting GnRH agonist/hMG protocol. PARTICIPANTS AND METHODS: One hundred eighty nine couples having their first trial of ICSI for male factor infertility were divided into two groups. Group I (no = 33) received CC 100-150 mg/day for five days starting from day 2 of the cycle and 150 IU of hMG/day on days 6-10. GnRH antagonist (Centrorelix) 0.25 mg/day was started when the leading follicle reached 16 mm in the absence of an LH surge. Group II (no = 156) received 0.1 mg Deacapeptyl/day as our standard long protocol. RESULTS: Clinical pregnancy was observed in 8 out of the 33 cases in group I (24%) while in group II, 92 out of 156 achieved clinical pregnancy (59%), the difference was statistically significant (P = 0.019). The cost of medications/cycle was estimated to be 1110+/-492 E.P in group I, while it was 1928+/-456 E.P. in group II. However, the total cost per pregnancy was 19653 EP in group I and 10047 EP in group II. CONCLUSION: The use of the clomid/hMG/antagonist protocol is not a cost effective strategy and should not be recommended in IVF-ICSI cycles. PMID- 12580840 TI - Cytogenetic studies in patients with reproductive failure. AB - BACKGROUND: Cytogenetic studies in patients with reproductive failure AIM: To investigate the contribution of chromosomal abnormalities in sub fertility and in couples with repeated abortions. METHODS: Hundred and 13 couples who had at least two or more spontaneous abortions and 65 women and 63 men with infertility were analyzed cytogenetically. RESULTS: Major chromosomal rearrangements were found in 8% and minor variants in 6% in the study population. Major chromosomal aberrations were judged to explain 4.9% of recurrent abortions and 13% of infertility. Chromosomal abnormalities in infertile men occurred in 5% and in infertile women in 21.5%. The chromosomal abnormalities were structural (57%), numerical (18%) or mosaics (25%). CONCLUSIONS: Chromosomal aberrations in recurrent abortions are mostly structural ones and those in female infertility mosaicism of sex chromosomes. Turner's syndrome, Turner variants and XY females are detected as a cause of female infertility. The structural and numerical aberrations of either sex or autosomal chromosomes were found in infertile men. PMID- 12580841 TI - Preoperative transcervical analgesia for laparoscopic sterilization with Filshie clips. A double blind randomized trial. AB - OBJECTIVE: To evaluate the use of preoperative transcervical analgesia for reducing postoperative pain from laparoscopic sterilization with Filshie clips. DESIGN: A double blind, randomized, placebo-controlled trial. SETTING: Gynecologic day-care clinic of Vejle Hospital, Vejle, Denmark. SAMPLE: Seventy eight healthy women were attending the public hospital requesting sterilization. METHODS: Participants received bupivacaine 0.25% 20 mL or placebo normal saline 20 mL injected into the uterus, though the fallopian tubes and into the pelvis before tubal occlusion with Filshie clips. MAIN OUTCOME MEASURES: Post-operative visual analog pain score and request for analgesia were recorded. RESULTS: At 30 min after clip application there was a non-significant reduction in pain-score on VAS. However, at 1, 2, 3 and 4 h median pain score on VAS was similar in both groups. Patients request for postoperative analgesia was similar in the two groups. CONCLUSIONS: Preoperative transcervical perfusion of the uterus, fallopian tubes and into the pelvis did not reduce postoperative pain or request for analgesics after laparoscopic sterilization with Filshie clips. PMID- 12580842 TI - Relationship between polycystic ovaries, body mass index and insulin resistance. AB - BACKGROUND: To investigate insulin levels in lean and overweight women with and without polycystic ovaries. An observational, cross-sectional study at The Northem General Hospital, Sheffield, UK. METHODS: Sixty-eight women born at Jessop Hospital, Sheffield, between 1952 and 1953 were divided into four groups according to the status of their polycystic ovaries and body mass index: either > or approximately 25. Therefore, this was an unselected sample, unlike previous studies that have recruited from endocrine clinics or similar. Subjects underwent pelvic ultrasonography to visualize their ovaries in order to diagnose or exclude polycystic ovaries. They all underwent a short insulin tolerance test. RESULTS: Women with a body mass index > 25 and with polycystic ovaries were the most insulin resistant. Women with a body mass index of < or = ?25 and with normal ovaries were the most insulin sensitive. Women with a body mass index < or = ?25 and polycystic ovaries were the more resistant than those with a body mass index > 25 and with normal ovaries. CONCLUSION: Obesity increases insulin resistance, and the presence of polycystic ovaries increases insulin resistance. The presence of polycystic ovaries appears to have a stronger influence than obesity on insulin resistance. This is the first study to demonstrate these relationships using unselected volunteers. PMID- 12580843 TI - Endometrial ablation with NovaSure GEA, a pilot study. AB - OBJECTIVE: A prospective, single-arm, multicenter, controlled, observational pilot study to assess the safety and efficacy of the NovaSure Global Endometrial Ablation (NovaSure GEA) system in women with severe menorrhagia secondary to dysfunctional uterine bleeding (DUB). SETTING: University tertiary referring center and central hospital. PATIENTS: Forty-six premenopausal women aged 41.3 years with menorrhagia secondary to DUB unresponsive to medical therapy, who had completed childbearing, with non-distorted uterine cavities were included in the study. METHODS: Bleeding score was recorded before, 6 and 12 months after treatment. All patients received the treatment under intravenous sedation with para-cervical block and were treated with Nova Sure endometrial ablation for mean 94 s at the day surgery unit. RESULTS: No serious intraoperative complications were observed. Treatment time averaged 94 s. Forty-six patients have completed 6 months of follow-up and 45 patients completed the 12-months follow-up. 50% of the patients were amenorrheic at 6 months follow-up and 58% had complete cessation of uterine bleeding (amenorrhea) at 12 months follow-up. CONCLUSIONS: Preliminary results indicates that the NovaSure GEA System is an effective treatment for DUB, achieving 58% amenorrhea after 12 months. PMID- 12580844 TI - Laser acupuncture and low-calorie diet during visceral obesity therapy after menopause. AB - BACKGROUND: To compare the efficiency of low-calorie diet and low-calorie diet applied together with laser acupuncture in the therapy of visceral obesity in postmenopausal women. METHODS: The study population consisted of 74 postmenopausal females with visceral obesity who were divided into two groups according to an employed 6-month slimming procedure. In the first group (n = 36) a low-calorie diet was applied, while women in the second group (n = 38) were on the same kind of diet, having additionally one cycle of laser acupuncture procedure at the same time. At baseline and at the end of the study, body weight, body mass index and waist-to-hip ratio were determined in all women. RESULTS: After 6 trial months both groups exhibited a statistically significant drop in body weight, body mass index and waist-to-hip ratio. The mean reduction of body weight, body mass index and waist-to-hip ratio was significantly higher in the second group of women (laser acupuncture plus low-calorie diet). CONCLUSIONS: (1) Our results testify that the combination of a low-calorie diet and laser acupuncture is characterized by a higher efficacy than a low-calorie diet alone in lowering body weight, body mass index and waist-to-hip-ratio. (2) Laser acupuncture is an additional useful healing method in the therapy of visceral postmenopausal obesity. PMID- 12580845 TI - Leiomyosarcoma of the uterus: a clinicopathologic study of 21 cases. AB - BACKGROUND: The purpose of the present study was to identify the prognostic factors of and to determine the most appropriate mode of treatment for uterine leiomyosarcoma. METHODS: We reviewed the hospital records, including surgical notes and pathologic reports, of 21 patients with uterine leiomyosarcoma treated at Chang Gung Memorial Hospital, Kaohsiung, Taiwan, between 1987 and 1997. Univariate analysis was performed using the log-rank test. Cox regression was used to identify independent prognostic factors. RESULTS: The mean follow-up time was 30 months, and the 5-year disease-free survival rate was 55%. Evaluating the correlation between clinicopathologic parameters and survival, early stage (p = 0.0002), tumor cells without necrosis (p = 0.0026), low-grade tumor (p = 0.015), absence of vascular space involvement (p = 0.006), and tumor without atypia (p = 0.016) were associated with good prognosis. However, in a multivariate analysis using the Cox model, only advanced stage (p = 0.032) and tumor necrosis (p = 0.032) were found to be independent poor prognostic factors. The 5-year disease free survival was only 21% in patients with the presence of any one or both of these two factors. Five of 11 patients in this group had received aggressive adjuvant therapy after surgery, and none of them survived more than 19 months. CONCLUSIONS: We found that patients with advanced stage or presence of tumor necrosis had an extremely poor prognosis. Adjuvant therapy seemed to play a limited role, and provided no survival benefit. Treatment for these patients should be palliative until effective therapeutic modalities prove otherwise. PMID- 12580846 TI - Quality improvement project in cervical cancer screening: practical measures for monitoring laboratory performance. AB - BACKGROUND: We conducted a quality improvement project in a cervical cancer screening programme in Helsinki in order to see if detection of precancerous lesions could be influenced by external (participation rate) and internal (laboratory praxis) quality measures. METHODS: In order to increase the participation rate, a second personal invitation to Pap-test was mailed to nonparticipants of the first call. In order to improve the quality of screening, the cytotechnicians monitored their performance longitudinally by recording the number of slides reviewed per day, the pick-up rate of abnormal smears, the report of the consulting cytopathologist, and the number of histologically verified lesions detected from the cases that they had screened. Regular sessions were held to compare the histological findings with the cytological findings of all cases referred for colposcopy. No pressure was applied on the cytotechnicians to ensure that they felt comfortable with their daily workload. RESULTS: A total of 110 000 smears were screened for cervical cancer at the Helsinki City Hospital during 1996-99. Initially, the overall participation rate increased from 62% to 71%. The number of histologically confirmed precancerous lesions (CIN 1-3) more than doubled and their detection rate increased from 0.32% to 0.72%. CONCLUSIONS: Continuous education and feedback from daily work performance were important, yet rather inexpensive means in increasing laboratory performance. Additional measures are needed to further increase the participation rate. Impact of the quality measures on cancer incidence needs to be assessed later on. PMID- 12580847 TI - Cesarean section and reoperative mitral valve replacement for thrombosis of a mechanical valve in a 32-week parturient. PMID- 12580848 TI - Lipogranuloma peritonealis caused by spontaneous rupture of a benign cystic ovarian teratoma. PMID- 12580849 TI - Domestic violence in late pregnancy and splenic injury: delayed presentation. PMID- 12580850 TI - Successful balloon tamponade of post cesarean hysterectomy pelvic bleeding: a case report. PMID- 12580851 TI - Is sleep disorder treatment appropriate for premenstrual syndrome? PMID- 12580855 TI - Increased hepatocyte growth factor level in cerebrospinal fluid in Alzheimer's disease. AB - BACKGROUND & OBJECTIVE: Hepatocyte growth factor (HGF), also known as the scatter factor, is a potent mitogen for mature hepatocytes, and also has multifunctional effects on some cells in various organs. Recently, we have found expression and localization of HGF in white matter astrocytes in human brain tissues. Furthermore, immunohistochemistry using anti-HGF antibody revealed more intense immunolabeling in Alzheimer's disease (AD) than control brains. The aim of the study is to investigate the level of HGF in cerebrospinal fluid (CSF) from patients with AD. MATERIAL AND METHODS: We examined the level of HGF in CSF from 34 AD and 15 age-matched disease control patients by highly sensitive enzyme linked immunoabsorbent assay (ELISA) system. RESULTS: Consistent with the immunohistochemical data, a significantly higher concentration of HGF in AD CSF was found as compared with controls. A significant correlation was also seen between CSF HGF levels and white matter high-signal foci determined on brain magnetic resonance imaging (MRI) in AD patients. CONCLUSION: These results indicate that CSF HGF levels correspond with the white matter damage in AD brain. PMID- 12580856 TI - The ideal characteristics of antiepileptic therapy: an overview of old and new AEDs. AB - New and improved anti-epileptic drugs (AEDs) have made the concept of choice, according to the individual prognosis and probable response to specific regimens, increasingly feasible. Inter-individual variability in syndrome severity and complexity make individualization necessary. We propose three categories of disorder control according to the individual objectives of the patient: (1) seizure control, (2) epilepsy control and ultimately, (3) "epilepsy cure"; the latter remaining a largely idealistic target today. An AED is likely to be successful if it exhibits "optimal" characteristics, such as drug efficacy, tolerability, pharmacokinetics, interactions and cost-effectiveness. This review discusses the "optimal" characteristics of add-on AEDs, which, in addition to seizure control, will contribute to the achievement of epilepsy control and therefore address the currently unmet clinical needs of epilepsy treatment. PMID- 12580857 TI - Microalbuminuria and hyperthermia independently predict long-term mortality in acute ischemic stroke patients. AB - OBJECTIVES: To investigate the association between microalbuminuria (MA) and hyperthermia in acute ischemic stroke and to evaluate their significance as the predictors of long-term mortality after stroke. MATERIAL AND METHODS: We assessed neurologic deficit, urinary albumin excretion and body temperature in 60 patients admitted within 24 h after the onset of their first ischemic stroke. Outcome was assessed by 90-day and 1-year mortality. RESULTS: MA was found in 46.7% of patients. Hyperthermia was found in 18.3% patients on Day 1 and in 25% patients on Day 2. The correlation between albuminuria on Day 2 and the body temperature on Days 1 and 2 was found (r = 0.45, and r = 0.30, respectively; both P < 0.05). The mortality was significantly higher in the group of patients with both MA and hyperthermia on Day 2 (73% vs 10% after 90 days; P < 0.0001 and 73% vs 18% after 1 year, P < 0.005). In the logistic regression analysis, albuminuria (P = 0.017), hyperthermia on Day 1 (P = 0.028) and neurologic deficit on admission (P = 0.044) independently predicted 1-year mortality after ischemic stroke. CONCLUSION: Daily urinary albumin excretion correlates with the body temperature in acute stroke patients, but the predictive power of both these variables is independent of that association. PMID- 12580858 TI - Does carpal canal stenosis predict outcome in women with carpal tunnel syndrome? AB - OBJECTIVE: We investigated the role of carpal canal stenosis as a predictor of outcome in patients who underwent surgical treatment for carpal tunnel syndrome (CTS). METHODS: We performed magnetic resonance imaging (MRI) of the wrist in 31 female patients with clinically and neurophysiologically confirmed CTS. With a computerized analyser we quantitatively calculated the focal narrowest point of the tunnel. Patient's assessment of CTS-related symptoms were obtained by using a visual analogue scale before, and 6 months after treatment. RESULTS: Seventeen (56%) patients improved in all symptoms after treatment. The focal narrowest point of the tunnel was identified at its distal third in all patients, at 8 mm from the outlet. The median area of the narrowest point in those who improved clinically (n=17) was 238.9 mm2 compared with 269.8 mm2 in others (n=14), P=0.046. CONCLUSION: Identification of carpal canal stenosis may be important in selecting candidates for treatment in symptomatic CTS. PMID- 12580859 TI - Effect of apolipoprotein E genotype on in-hospital mortality following intracerebral haemorrhage. AB - OBJECTIVE: To determine the relationship between the apolipoprotein E (APOE) epsilon4 allele and in-hospital mortality from intracerebral haemorrhage (ICH). MATERIAL AND METHODS: Patients admitted to two acute stroke units with ICH were prospectively evaluated and APOE genotyped. In-hospital survival was recorded in 176 patients. RESULTS: There were 85 men and 91 women, mean age 68 years. Fifty two (30%) of the 176 patients died in hospital. After adjusting for sex, age, hospital, and race, increased age (P = 0.009) and the presence of the APOEepsilon4 allele (P = 0.026) significantly reduced in-hospital survival. CONCLUSION: The APOEepsilon4 allele in this population may be associated with poor survival following ICH. PMID- 12580860 TI - Coagulopathy and outcome in patients with chronic subdural haematoma. AB - OBJECTIVE: The coincidence of coagulatopathy and chronic subdural haematoma (CSH) requires correction of coagulation to facilitate surgery. We investigated the correlation between coagulopathy and outcome in CSH patients. MATERIAL AND METHODS: We analysed past medical history, surgical treatment and coagulation parameters of 114 patients. RESULTS: Coagulation disorders were found in 42%. Preoperative treatment with prothrombin complex concentrate was necessary in 14%. A significant difference (P < 0.05) of the preoperative level of platelets was found between recurrent CSH and non-recurrent group. Totally, we had to perform re-operations in 17.5%. Eighty-one patients presented with Glasgow coma scale (GCS) > or = 13. After surgery GCS was > or = 13 in n = 92. There was an improvement of GCS in 46 cases, 61 patients maintained GCS score levels. Outcome was significantly worse in the alcoholic group (P < 0.001), and in the recurrent group (P < 0.05). In patients with substitution of coagulation factors, outcome was worse in the group with post-operative substitution only (P < 0.05). CONCLUSION: In CSH, the coagulation parameters and a subtle correction of coagulation are of special interest, regarding the worse outcome in patients with recurrent CSH and in those requiring post-operative substitution. PMID- 12580861 TI - VNS in patients with previous unsuccessful resective epilepsy surgery: antiepileptic and psychotropic effects. AB - OBJECTIVES: To assess the efficacy of vagus nerve stimulation (VNS) in patients with medically and surgically intractable complex partial seizures (CPS). PATIENTS AND METHODS: Sixteen patients with previous temporal [15] and frontal [one] resections were treated with VNS between 1994 and 1999 at King's College Hospital, London, UK. Post-operative video-electroencephalogram telemetry had shown that CPS started from the operated side in 12 patients, contralaterally in three and bilaterally independently in one. RESULTS: Three patients (18.75%) had 50% or more reduction in seizure frequency, but one showed severe worsening of epilepsy, which remitted upon VNS discontinuation. The antiepileptic effect of VNS was not different with respect to the type of operation (anterior temporal lobectomy vs amygdalohippocampectomy), the side of operation, or the side of seizure onset. We observed psychotropic effects in two patients with post-ictal psychosis, in two others with depression, and in a child with severe behavioral disorder. CONCLUSIONS: VNS may have a rather limited antiepileptic role to play in patients with persistent seizures following epilepsy surgery, but may independently possess useful antipsychotic and mood-stabilizing properties. PMID- 12580862 TI - HHV-6 DNAaemia in patients with multiple sclerosis in Kuwait. AB - OBJECTIVE: To study the presence of active human herpesvirus type-6 (HHV-6) infection indicated by the presence of HHV-6 DNA in serum (DNAaemia) in patients suffering from multiple sclerosis (MS) in Kuwait. MATERIAL AND METHODS: Sera from 24 patients with MS (18 relapsing-remitting, six secondary progressive disease), control sera from 13 patients suffering from other neurological diseases and sera from 20 healthy volunteers were examined for the presence of HHV-6 DNA by nested polymerase chain reaction (PCR) test. RESULTS: None of the MS patients, nor patient controls were positive for HHV-6 DNAaemia while, one of the normal healthy controls was positive for HHV-6 DNAaemia. CONCLUSION: HHV-6 DNAaemia, indicating active HHV-6 infection could not be demonstrated in a sample of Kuwaiti patients suffering from active clinically definite MS. There is no evidence to incriminate this virus in the pathogenesis of MS in Kuwait. PMID- 12580863 TI - Leisure time and social activity after a newly diagnosed unprovoked epileptic seizure in adult age. A population-based case-referent study. AB - PURPOSE: To investigate leisure time activity and factors that influence it in an adult cohort having newly diagnosed unprovoked epileptic seizures. PATIENTS AND METHODS: A cohort of 62 patients > or =17 years with a newly diagnosed unprovoked epileptic seizure during 1985-87 and 93 sex- and age-matched referents were followed up regarding leisure time activity, marital status and driver's license for 10 years till 1996. Data were questionnaire derived. RESULTS: Patients became significantly less physically active, traveled abroad less often and were generally less active during their leisure time than referents. No other significant differences were observed. CONCLUSION: After a newly diagnosed unprovoked epileptic seizure in adult age, most leisure time activities, marital status and holding of driver's license are not affected. Patients become physically less active than referents. The role of seizures for this difference is unclear and other medical and social factors should be considered. PMID- 12580864 TI - The expression of adhesion molecules in muscle biopsies: the LFA-1/VLA-4 ratio in polymyositis. AB - OBJECTIVES: The expression of three pairs of adhesion receptors and ligands was examined in 22 consecutive muscle biopsies showing morphological signs of inflammation. MATERIAL AND METHODS: The following groups were studied: patients with polymyositis (PM) (n=7), patients with myositis that did not fulfil criteria for PM, i.e. suspected PM (n=5), patients with other diseases, with no clinical signs of inflammatory myopathy (n=6), and a small group of non-PM inflammatory myopathies (n=4). The endothelial expression of ICAM-1, VCAM-1 and E-selectin was evaluated, as was the cellular expression of LFA-1, VLA-4 and SLex. In addition, the expression of MHC class I and II was studied. RESULTS: The ratio between the number of cells expressing LFA-1 and VLA-4 showed significant differences between the groups, with the lowest values in PM. CONCLUSION: The LFA-1/VLA-4 ratio should be suitable for diagnostic purposes. Our findings also indicate that the VLA-4/VCAM-1 system is important for chronic T cell inflammation in muscle, in line with findings in other "hidden" organs like joints and the central nervous system. PMID- 12580866 TI - Case ascertainment in stroke studies: the risk of selection bias. AB - OBJECTIVES: The purpose was to compare the completeness of case ascertainment in two stroke registers, one local population-based, the other a national quality register (Riks-Stroke), and to examine if patient characteristics could be affected by selection bias. MATERIAL AND METHODS: By the way of linking and matching computer files, the completeness of case ascertainment was evaluated. RESULTS: In the local stroke incidence study 377 patients were included. Of them, 63% were reported to the hospital-based national quality register. The case fatality was lower in the national register. A larger proportion of the patients in the national register appeared to have been treated in a stroke unit and undergone rehabilitation, and computerized tomography seemed to have been performed in a larger proportion. CONCLUSIONS: Because of selection bias, outcome data get skewed when case ascertainment does not embrace all stroke cases. A community-based stroke register is the golden standard when measuring stroke incidence. PMID- 12580865 TI - Status epilepticus as a manifestation of hepatic encephalopathy. AB - OBJECTIVES: Seizures have been described as a rare manifestation of hepatic encephalopathy. MATERIAL AND METHODS: We present a 54-year-old female, with 6 year history of decompensated, hepatitis B liver cirrhosis, admitted with generalized seizures. She reported a history of recurrent episodes of hepatic encephalopathy, spontaneous bacterial peritonitis, tense ascites and variceal hemorrhage. Neurologic examination revealed a comatose patient, without papilledema. Laboratory examinations were suitable with cirrhosis and mild renal failure. Blood gas examination revealed severe metabolic acidosis and hypoxemia. Plasma NH3+ levels upon admission were twice normal. Brain computed tomography and magnetic resonance imaging were normal. Electroencephalogram showed diffuse sharp waves, consistent with hepatic encephalopathy, grades III-IV. RESULTS: Status epilepticus was refractory to continuous antiepileptic treatment. However, it was resolved after 24-h therapy with lactulose. Blood NH3 levels were simultaneously normalized with clinical improvement. CONCLUSIONS: We consider the status epilepticus of our patient to be a rare manifestation of hepatic encephalopathy. PMID- 12580867 TI - Platelet serotonin transporter in stroke patients. AB - OBJECTIVES: Post-stroke depression can be treated with serotonin transport inhibitors suggesting a role for the serotonin system in these patients. The number of platelet serotonin transporters in stroke patients and in control subjects have been measured in this study. MATERIAL AND METHODS: Newly admitted stroke patients who did develop or who did not develop a post-stroke depression, non-acute patients who previously had had a stroke and control subjects were compared. The number of platelet serotonin transporters was analysed by ligand binding methodology. RESULTS: The number of platelet serotonin transporters was low shortly after a stroke compared with normal subjects; no difference was found between the stroke patients who developed a post-stroke depression and those who did not. CONCLUSION: A low number of platelet serotonin transporters may be a non specific state marker for a condition as acute stroke. PMID- 12580868 TI - Multiple sclerosis, interferon beta and clinical thyroid dysfunction. AB - The objective of this study was to investigate frequency and presentation of clinical thyroid dysfunction in patients treated with interferon beta (IFN-beta). We have collected the cases of clinical thyroid dysfunction in 700 consecutive patients receiving IFN-beta for multiple sclerosis (MS). Five patients (four women, one man) treated with IFN-beta1b developed hyperthyroidism. Three of them have secondary progressive MS, and two have relapsing-remitting MS. It was necessary to stop IFN-beta in three cases; these patients still require carbimazole after several months. In the two other cases, hyperthyroidism disappeared spontaneously. Two patients (one man and one woman) treated with IFN beta1a developed hypothyroidism. One of them required l-thyroxine. Lastly, an increased thyroid volume without modification of thyroid hormones plasma levels was discovered in a patient receiving IFN-beta1a. Among patients treated with IFN beta, clinical thyroid dysfunction is much rarer than laboratory thyroid dysfunction. However, this side-effect is sometimes severe. PMID- 12580869 TI - Antimigraine drug consumption in Spain (1990-2000). AB - OBJECTIVES: New marketed antimigraine drugs (triptans) are promising in the management of migraine pain. The impact of these new drugs on the overall consumption of the antimigraine drugs merits an analysis. MATERIAL AND METHODS: The package units of antimigraine drugs, sold and reimbursed by the Spanish National Health System, are used for estimations. Data are presented as defined daily doses per 10,000 inhabitants/day. RESULTS: During the 1990-2000 period, antimigraine drug utilization as a whole decreased by more than 25%; triptan use increased while ergot alkaloids' declined. CONCLUSIONS: The pattern of antimigraine drug utilization in Spain has changed noticeably in accordance with recent recommendations. PMID- 12580870 TI - Severe amphethamine-induced bruxism: treatment with botulinum toxin. AB - While chewing and grinding movements have been observed in amphetamine addicts, recognition and management of this problem have rarely been highlighted. Botulinum toxin (BTX) has previously been demonstrated to be effective for bruxism associated with movement disorders, such as cranial-cervical dystonia. However, there is little information on its use in tardive bruxism. Here we report an amphetamine addict who presented with medically intractable bruxism, and discuss its pathophysiology and successful treatment with BTX. PMID- 12580871 TI - Developments in radiotherapy and adjuvant chemotherapy for head and neck cancer. PMID- 12580872 TI - Hair cell regeneration in the inner ear: a review. AB - Hair cell regeneration has been shown to occur in the inner ear of mammals. Specifically, it has been demonstrated in the vestibular system and not the organ of Corti. Recent evidence suggests that the degree of the regenerative response may be augmented pharmacologically. This review discusses the field of hair cell regeneration in fish, amphibians, birds and mammals, and the relationship of regeneration to functional recovery PMID- 12580873 TI - A comparative study of the optical characteristics of commonly used sinoscopes: do you know where you are looking? AB - Endoscopic ethmoidectomy is a delicate operation on the thin, honey-comb-like laminae of the ethmoidal sinuses. A high degree of precision is required to minimize disruption of the natural sinus mucociliary clearance system and to reduce the risk of orbital and skull base complications. It is therefore important that surgeons are aware of variation in the optical characteristics of the rod-lens telescopes produced by different manufacturers. A simple but precision endoscope goniometer was designed and manufactured. This was used to carry out a comparative study of the sinus telescopes in common use. The results showed significant variation in the view angle of the four brands of telescopes (Karl Storz, Olympus, Richards Smith & Nephew, and Richard Wolf) tested. Differences in the position of the horizon in the visual field, and different degrees of visuospatial distortion were evident. We also noted and measured the blind angle of telescopes, which, by virtue of a high degree of angulation, cannot view the horizon. PMID- 12580874 TI - A comparison of physician and patient perception of the problems of habitual snoring. AB - Assumptions are often made by doctors about the nature of the impact of uncomplicated snoring. The aim of this study was to provide a comprehensive view of the problems experienced by snoring patients, and to compare these with the perceptions of otolaryngologists. Part 1: 121 snorers and 419 otolaryngologists completed open-ended problems lists; Part 2: 56 snorers and 63 matched controls completed a mood questionnaire and the Golombok Rust Inventory of Marital State (GRIMS.) There was a clear agreement between snorers and otolaryngologists that sleep-related difficulties comprise the most frequent category of problems that may result from snoring. Snorers report a significantly greater number of physical problems and significantly fewer relationship problems than perceived by otolaryngologists. GRIMS scores were similar in patients and controls. Snorers, however, were significantly more depressed. Doctors appear to address snoring from the twin perspectives of damaged relationships and possible sleep apnoea. Their patients are more concerned about disturbing the sleep of others, and are subject to low mood. The snorers' 25 different responses are now being used as the basis for a Snoring Symptom Index. PMID- 12580875 TI - Sublingual packing as an useful adjunct to submandibular gland excision. AB - We describe a simple preoperative procedure that we have found of great benefit in submandibular gland excision. PMID- 12580876 TI - Laser Doppler flux-metry in laryngeal squamous cell carcinoma. AB - Tumour angiogenesis has recently attracted a great deal of attention as a critical part of oncogenesis and a necessary prerequisite for a malignant phenotype. Novel antiangiogenic therapy for solid tumours including laryngeal cancer is entering clinical trials. Quantifying microvessel density is considered the gold standard for measuring baseline angiogenesis and indeed 'the response to intervention'. We hypothesize that laser Doppler flux-metry could provide a non invasive reliable method of quantifying blood flux within tumours. The aims were to determine whether a laser Doppler flux meter could be used as a reliable and reproducible method of estimating blood flux in the human larynx and to establish baseline Doppler flux recordings for the human larynx. The method used was a validation study in patients with laryngeal squamous cell cancer and normal controls. Statistical analysis was performed using SPSS software. We have demonstrated good reproducibility of laser Doppler measurements in human laryngeal mucosa (correlation coefficient 0.956 @P = 0.01). We have also derived arbitrary means of laser Doppler flux-metry in normal laryngeal mucosa and in squamous cell carcinoma of the larynx. Comparisons between normal and tumour laser Doppler flux-metry (LDF) readings showed no significant difference. We suggest that Laser Doppler flux-metry is a potentially useful tool with which to study blood flow in the larynx and propose arbitrary LDF levels for the normal and diseased human larynx. PMID- 12580877 TI - Endoscopic repair of nasal septal perforation with acellular human dermal allograft and an inferior turbinate flap. AB - Nasal septal perforations present a distinct challenge to the otolaryngologist and a significant cause of symptoms to affected patients. Many surgical techniques for the repair of septal perforations have been described. Connective tissue autografts are commonly used as interpositional grafts between the septal flaps. Recently acellular human dermal allograft has been used with success. In total, 17 patients with symptomatic anterior nasal septal perforations that had failed conservative treatment underwent a closed endoscopic repair of their perforations with acellular human dermal allograft (alloderm) and an anteriorly based inferior turbinate flap; 13 patients had a successful closure of the perforation, two patients, despite initial success, re-perforated as a result of persistent crust picking and, in two patients, the graft failed. With appropriate patient selection and stringent postoperative care the authors consider this technique offers a good surgical outcome for the closure of septal perforations. PMID- 12580878 TI - Assessment of a cytology grading system for head and neck masses. AB - A grading system for Fine Needle Aspiration Biopsy Cytology (FNAC) has been in routine use in Peterborough since 1994 for reporting on fine needle aspiration biopsies of neck lumps. The records of all 276 patients who had neck lump fine needle aspiration biopsies performed between 1994 and 1999 were reviewed retrospectively. The 193 patients who had definitive histology available were included in the study of whom 154 (80.2%) were considered adequate for grading. A learning curve for adequacy of specimens obtained is demonstrated, with adequacy rate increasing with increasing experience or seniority of the sampler. The grading system produced an overall accuracy rate of 73.3% with a predictive value for malignancy of 88.6% and a predictive value for benign conditions of 67.3%. Individual grades were assessed for predictive value of benign or malignant disease. Grade 5 proved completely reliable, with a predictive value for malignancy of 100%. The shortcomings of the other grades and the difficulties in diagnosing lymphoma by FNAC are discussed. PMID- 12580879 TI - Influence of prednisolone on the secretion of mucin from the HT29-MTX cell line. AB - Glucocorticoids have been used in the treatment of otitis media with effusion with promising but inconsistent results. The HT29-MTX cell line is a completely differentiated and almost exclusively mucus-secreting goblet cell line. To assess the potential of steroids in suppressing mucin secretion, we have studied the response of this cell culture to prednisolone. Confluent cell cultures were trypsinized, subcultured in six-well plates and incubated with five doses of prednisolone from 10-3 M to 10-11 M and over a varying time course from 6 to 36 h. Analysis was performed using a monoclonal mouse antibody to human gastric mucin by dot-blot ELISA. Prednisolone caused a consistent reduction in mucin production from this cell line. Increasing concentrations of prednisolone resulted in increasing suppression of MUC5AC secretion. There is a dose-dependent suppression of mucin secretion by prednisolone, with a maximum effect of 21% over control seen at the highest steroid concentration used. PMID- 12580880 TI - Can the Sino-nasal Outcome Test (SNOT-22) be used as a reliable outcome measure for successful septal surgery? AB - Septoplasty and submucous resection are common procedures in the UK. This study looks prospectively at 40 patients undergoing surgery at two hospitals. A pre- and postoperative assessment (3 months) was made using the Sino-nasal Outcome Test. This test was originally designed for rhinosinusitis but our study suggests that it is a useful tool in nasal septal surgery, in that it combines both nasal specific and general health questions, which can be analysed individually or together. Improvements in nasal obstruction (75%), facial pain (33%) and catarrh (10%) were noted. PMID- 12580881 TI - The admission of patients with peri-tonsillar abscess to a general ENT ward and its influence on secondary post-tonsillectomy haemorrhage. AB - Patients with peri-tonsillar abscess require hospitalization. The aim of this prospective study was to determine whether the admission of patients with peri tonsillar abscess to the same ward as those undergoing tonsillectomy influences the incidence of secondary post-tonsillectomy haemorrhage. We included all adult patients undergoing in-patient tonsillectomy (n = 183) and all patients with proven peri-tonsillar abscess (n = 46) over a 1-year period. Both patient groups were nursed in the same general otolaryngology ward. The operation dates of those patients who developed secondary post-tonsillectomy haemorrhage were matched to the dates when patients with peri-tonsillar abscess were admitted. Out of the 71 patients who had tonsillectomy on days when patients with peri-tonsillar abscess were admitted, 10 developed secondary haemorrhage. Out of the 112 patients who had tonsillectomy on days when no one in the ward had peri-tonsillar abscess, 16 developed secondary haemorrhage. No difference was demonstrated (chi2 = 0.0014, P > 0.05). In this study, admission of patients with peri-tonsillar abscess to a general otolaryngology ward does not influence the incidence of secondary post tonsillectomy haemorrhage in patients undergoing elective surgery. PMID- 12580882 TI - Effect of nitric oxide donation on mucin production in vitro. AB - Otitis media with effusion (OME) is characterized by the accumulation of a viscous fluid rich in mucins in the middle ear cleft. There is increasing evidence that this fluid is the result of an inflammatory reaction and that nitric oxide (NO) is an important mediator in this reaction. The goblet cell line HT29-MTX produces principally MUC5AC, an important mucin in middle ear effusions, and thus is a good model for the study of mucus-secreting epithelia. Confluent cell cultures were trypsinized, subcultured and incubated with isosorbide dinitrate (ISDN), a NO donor, for 0.5, 1 and 2 h at a concentration of 1 mm and in concentrations of 0.01, 0.1, 0.5, 1 and 2 mm for 1 h. Experiments were performed four times. Mucin production was detected by a slot blot ELISA assay, using a monoclonal mouse antibody to human MUC5AC mucin. Statistical significance was tested using a one-way analysis of variance. NO donation by ISDN caused a consistent rise in mucin production above control. Maximal mucin production of 35% above control occurred at 1 h with 1 mm ISDN. Mucin production increased from 12% above control with 0.1 mm ISDN dinitrate to 45% above baseline with 2 mm ISDN. NO donation by ISDN results in an increase in mucus production, which is both dose and time related. This adds further evidence to an inflammatory model for mucus secretion in OME. PMID- 12580883 TI - Local and systemic eosinophilia in patients undergoing endoscopic sinus surgery for chronic rhinosinusitis with and without polyposis. AB - Eosinophils play a key role in the pathophysiology of sinonasal polyposis; their role in chronic rhinosinusitis without polyposis is less clear. To investigate this further, we biopsied diseased sinonasal tissue from 116 patients undergoing endoscopic sinus surgery and normal nasal mucosa from 24 controls. The patients were grouped, according to the nasal endoscopic appearance of their disease, into four clinical groups: chronic rhinosinusitis with no polyps, grade 1 polyps, grade 2 polyps, and grade 3 polyps. We also measured the peripheral blood eosinophil count. Histological analysis of the inflammatory cell content of the biopsies was carried out. The percentage of eosinophils in tissue from each of the clinical groups was significantly higher than that from the controls. There was no significant difference between the percentage of eosinophils in any of the clinical groups, but a trend. Peripheral eosinophil count increased with increasing severity of nasal disease. PMID- 12580885 TI - The future of Clinical and Experimental Ophthalmology. PMID- 12580886 TI - Cataract blindness: performance, priorities, politics. PMID- 12580887 TI - Diabetic retinopathy: yet another reason for a comprehensive eye-care programme for Australian Aborigines and Torres Strait Islanders. PMID- 12580888 TI - Current trends in the treatment of ocular melanoma by radiotherapy. AB - The Collaborative Ocular Melanoma Study (COMS) has recently confirmed once and for all that it is safe to attempt to preserve an eye with a posterior uveal melanoma by demonstrating no survival advantage of enucleation over plaque radiotherapy. While COMS has been under way, we have set out in London to define the selection criteria for conservative therapy versus enucleation for the various categories of melanoma in terms of size, location within the eye, and presence or absence of retinal detachment. The evolution of this approach has culminated in an overall ocular survival rate of 94% in 597 patients following radiation therapy combined with a mean loss of visual acuity of only 2.4 Snellen lines in the eyes preserved. PMID- 12580889 TI - Citation analysis and journal impact factors in ophthalmology and vision science journals. AB - Citation analysis has evolved over the last 50 years as one parameter for assessing the quality of research published in scientific, technology and social science journals. This is based on the assumption that influential research is widely cited by other scientists and clinicians. With the advent of the Internet, Journal Citation Reports from the Institute for Scientific Information (ISI-JCR) have become widely available to individuals and institutions. In an increasingly competitive research environ-ment, aspects of citation analysis have been suggested as simple proxy, objective measures to evaluate the research quality of a journal, published articles, research institutions and even individual researchers. This review article provides an overview of citation analysis, including definitions, uses of these reports, and related controversies and potential abuses. As it has become the most commonly used indicator, there is a particular focus on the use of the Journal Impact Factor (JIF). This is a widely quoted measure indicating the frequency with which the average article published in a journal of interest will be quoted within a specified time frame that therefore allows approximate comparisons of journals within a particular field of interest. Given the relative paucity of information in this area, emphasis is placed on citation analysis within ophthalmology, in particular in regard to the 43 ophthal-mology, vision science and optometry journals that are listed in the ISI-JCR 2001 reports. PMID- 12580890 TI - Outcome and number of cataract surgeries in India: policy issues for blindness control. AB - PURPOSE: To assess what impact attention to quality of cataract surgery and postoperative follow up can have on cataract blindness in India, and to estimate the number of surgeries needed to eliminate cataract blindness in India. METHODS: In a population-based sample of 5268 persons in two rural areas in the state of Andhra Pradesh, India, data on the visual outcome of cataract surgery were obtained on 129 eyes of 106 persons operated previously. These were compared with the visual outcome of cataract surgery performed during 1999 in 2394 eyes of 2213 persons at two new rural eye centres set up in these areas that pay particular attention to the quality of eye care. Blindness in an eye was defined as presenting distance visual acuity less than 6/60, and in a person as this acuity level in the better eye. These data were extrapolated to India. RESULTS: In the population-based sample, of the 129 operated eyes, 51 (39.5%) were blind after surgery, which included 41 (31.8%) from cataract surgery-related causes. Of the 106 persons in the population sample who had had cataract surgery in one or both eyes, 26 (24.5%) were still blind. In contrast, for the cataract surgery at the two rural eye centres paying attention to quality, 3.1% of the eyes and 1.8% of the persons were blind soon after surgery, but these figures could increase 2.6 times over the lifetime of these persons. The 3.5 million cataract surgeries in India in 2000 are estimated to result in 0.32 million persons having blindness averted over their lifetime. To eliminate cataract blindness in India, an estimated 9 million good-quality cataract surgeries are needed every year during 2001-2005, increasing to over 14 million surgeries needed every year during 2016 2020 on persons most likely to go blind from cataract. CONCLUSIONS: The number of persons in whom blindness is being averted due to cataract surgery in India is currently a very small fraction of the number blind from cataract. If adequate attention is given to the quality of eye care, for the current number of cataract surgeries in India it should be possible to increase by threefold the number in whom blindness can be averted with cataract surgery. In addition to improving quality, the annual number of cataract surgeries has to be at least threefold the current number during 2006-2010 if cataract blindness in India is to be eliminated. PMID- 12580891 TI - Prevalence of diabetic retinopathy in an Aboriginal Australian population: results from the Katherine Region Diabetic Retinopathy Study (KRDRS). Report no. 1. AB - BACKGROUND: The Katherine Region Diabetic Retinopathy Study (KRDRS) was carried out in the Lower Top End of the Northern Territory of Australia between 1993 and 1996 as part of the Northern Territory Eye Health Program. It investigated diabetic eye conditions and its determinants in the Aboriginal Australian population of the region. METHODS: The study provided cross-sectional data about diabetic retinopathy in the Aboriginal diabetic population of the Katherine region in 1993 and in 1996. A total of 234 people with diabetes were examined in 1993 and 243 in 1996. RESULTS: The prevalence of retinopathy was 18% in 1993 and 21% in 1996. The respective findings for maculopathy were 13% and 10%, for clinically significant macula oedema 8% and 6%, for proliferative stage subject retinopathy 0.9 and 1.3% and for vision-threatening retinopathy 8.5% and 6.7%. In 1993 the prevalence of diabetic retinopathy was 18% in the four major centres compared with 16% in the smaller communities. The findings for 1996 were 25% and 13%, respectively. In 1993 the prevalence of vision-threatening retinopathy was 8% in major centres compared with 7% in other communities. The findings for 1996 were 7% and 8%, respectively. Sex was not significantly related to the prevalence of retinopathy. CONCLUSION: Diabetic retinopathy is as much a problem in Aboriginal communities as in the wider Australian population but presents a unique set of challenges for health services and for clinicians. PMID- 12580893 TI - Pneumococcal keratitis: a clinical profile. AB - AIM: To study the clinical features of pneumococcal keratitis and response to ciprofloxacin therapy. METHODS: A retrospective analysis was undertaken of 58 patients with culture-proven pneumococcal keratitis seen over a period of 2 years. RESULTS: Pneumococcal keratitis accounted for 33.3% of bacterial keratitis. Most cases presented with non-severe keratitis (77.5%). Co-existing sac pathology was more frequent in pneumococcal ulcers as compared to non pneumococcal bacterial ulcers (50%vs 9%, P < 0.001). Characteristic clinical features enabling an accurate clinical diagnosis were found in 27.5% and lanceolate diplococci on Gram's stain were identified in 76% of cases. In vitro testing showed a high susceptibility to cephazolin and ciprofloxacin. All patients received ciprofloxacin as first-line therapy. Eighty per cent responded well with complete healing of the ulcer. A second drug was required in 8.5%. CONCLUSION: Ciprofloxacin therapy can be effective in the treatment of pneumococcal keratitis. PMID- 12580892 TI - Penetration of oral and topical ciprofloxacin into the aqueous humour. AB - AIMS: To investigate the penetration of ciprofloxacin via different modes of administration into the aqueous humour using capillary zone electrophoresis and to determine its prophylactic role in ophthalmic surgery. METHODS: A prospective study was conducted of 84 patients, divided into two groups, undergoing routine cataract surgery and intraocular lens implantation. Forty patients received 750 mg ciprofloxacin orally the evening before and on the morning of surgery 12 h apart. Forty-four patients received one drop of 0.3% topical ciprofloxacin 90 and 30 min prior to surgery. At the commencement of surgery 0.1-0.2 mL of aqueous fluid was sampled from the anterior chamber and assayed for ciprofloxacin concentration using capillary-zone electrophoresis. RESULTS: The concentration of ciprofloxacin in the group receiving topical doses was less than 0.1 micro g/mL, which is well below the minimum inhibitory concentration for cipro-floxacin inhibiting 90% (MIC90) of strains of Staphylococcus epidermidis (0.4 micro g/mL). The mean concentration of ciprofloxacin in the aqueous humour of the oral group was 0.26 micro g/mL with a standard deviation of 0.12 micro g/mL (range 0.09-0.67 micro g/mL) with only 12.5% achieving levels higher than MIC90 for S. epidermidis. CONCLUSION: The topical ciprofloxacin 0.3% as given in this study achieved poor aqueous humour concentration. Oral ciprofloxacin given in this regime should not on its own be considered adequate prophylaxis in intraocular surgery. Capillary zone electrophoresis can be used as an alternative tool to the existing high-pressure liquid chromatography methods for analysing ciprofloxacin level in the aqueous humour. PMID- 12580894 TI - Contact lens fitting for post-radial-keratotomy residual myopia. AB - PURPOSE: To formulate guidelines for contact lens fitting following radial keratotomy and to achieve a relationship between contact lens fitting parameters and refractive and keratometric parameters. METHODS: Thirty-five eyes of 30 patients who had post-radial-keratotomy residual myopia were evaluated for contact lens fitting. The base curve was selected by 'hit and trial' method until a proper fit was achieved, following which an over-refraction was done and final power of the contact lens obtained. RESULTS: The mean contact lens base curve was 43.04 +/- 1.62 D, which was steeper than the postoperative keratometry by 2.35 +/ 0.74 D. The base curve and the postoperative keratometry were correlated by regression analysis (r = 0.895, P < 0.001). The mean overall diameter of the contact lenses was 9.64 +/- 0.08 mm. The mean contact lens power was 5.81 +/- 3.07 D, which was more myopic than the residual myopia by 2.79 +/- 1.67 D. Regression analysis showed definite correlation between the contact lens power and postoperative myopia (r = 0.854, P < 0.001). CONCLUSION: For correction of post-radial-keratotomy residual myopia, contact lens fitting parameters can be determined by a regression formula using keratometric and refractive values. PMID- 12580895 TI - Randomized clinical trial of the efficacy and safety of tropicamide and phenylephrine in preoperative mydriasis for phacoemulsification. AB - PURPOSE: To compare the mydriatic effect and safety between different concentrations of tropicamide and phenyle-phrine in preoperative mydriasis for phaco-emulsification. METHODS: Two hundred and seventeen consecutive eyes in the same number of Chinese patients undergoing phaco-emulsification under local or topical anaesthesia in a university-based eye hospital were analyzed. Patients were randomized into two groups by cluster randomization, each group receiving a different preoperative mydriatic regimen. Regimen A consisted of tropicamide 1.0% with phenylephrine 2.5%, and Regimen B consisted of tropicamide 0.5% with phenylephrine 0.5%. The main outcome measures were horizontal pupillary diameter, systolic, diastolic and pulse pressure and pulse rate. RESULTS: The group who received Regimen A attained a mean horizontal pupillary diameter of 7.00 +/- 1.06 mm. Their pupils were significantly larger than those receiving Regimen B (6.61 +/- 1.03 mm, P = 0.007). No untoward cardiovascular effects were noted in either groups. CONCLUSION: Regimen A attained better preoperative mydriasis for phacoemulsification than Regimen B. Both regimens were safe with regard to their cardiovascular effects. The combination of tropicamide 1.0% and phenylephrine 2.5% is recommended as preoperative mydriatic for phacoemulsification in Chinese patients who have darkly pigmented irides. PMID- 12580896 TI - Questioning the need for routine bone marrow aspiration and lumbar puncture in patients with retinoblastoma. AB - PURPOSE: This study assesses the value of routinely investigating children with retinoblastoma with bone marrow aspiration and lumbar puncture, staging investigations not without risk and trauma to the patient, emotional stress on parents and financial cost to the community. METHODS: Medical files and specimens were obtained and examined for patients with retinoblastoma presenting to the Royal Alexandra Hospital for Children, Camperdown and the Children's Hospital at Westmead, Sydney, from 1975 to 2001. RESULTS: In total, 123 patients presented; 62 (50.4%) were boys and 61 (49.6%) were girls. Of these 123 patients, 74 (60.2%) had unilateral disease, 46 (37.4%) involving the left eye and 28 (22.8%) involving the right eye. There were 47 (38.2%) patients with bilateral disease, and two (1.6%) with trilateral disease. Mean age of presentation was 17.9 months (23.1 months for unilateral subjects; 10.3 months for bi-lateral subjects; 3.5 months for trilateral subjects). There were 13 (10.6%) with a positive family history. Of 74 unilateral subjects, 70 (94.6%) required enucleation and four (5.4%) were salvaged. Of 47 bilateral subjects, 13 (27.7%) did not require enucleation, 27 (57.4%) required unilateral enucleation and seven (14.9%) required bilateral enucleation. Both trilateral subjects died. Of all 123 subjects, 112 (91.1%) had bone marrow aspiration and lumbar puncture performed during initial assessment, and none showed evidence of malignancy. CONCLUSIONS: Given the small but significant risks associated with these procedures, the results of this study cannot support bone marrow aspiration and lumbar puncture as routine investigations in all patients presenting with retinoblastoma, suggesting a more limited usage of these investigations is warranted. PMID- 12580898 TI - Preventative effect of deferoxamine on degenerative changes in the optic nerve in experimental retrobulbar haematoma. AB - PURPOSE: Changes in the optic nerve due to the breakdown of blood elements and the effect of deferoxamine on these changes were evaluated. METHODS: The study group consisted of 12 rabbits. Three rabbits were used as control. In Group I, three rabbits had bilateral retrobulbar haematoma induced. In Group II, three rabbits had bilateral retrobulbar haematoma induced and were given systemic deferoxamine. In Group III, three rabbits were only given deferoxamine. At day 21, all the rabbits underwent bilateral enucleation and specimens from the orbital fat tissue were removed for light microscopic examination. The optic nerves were examined by light and electron microscopy. RESULTS: Light microscopic examination of the optic nerves did not reveal significant differences among the groups. Electron microscopy revealed ultrastructural changes in Group I. There were no significant pathological findings observed in Groups II or III. There was abundant iron pigment in the orbital fat tissue of Group I, but only a small amount in Group II. CONCLUSION: Systemic deferoxamine treatment is effective in preventing degenerative changes in retrobulbar haema-toma. PMID- 12580897 TI - Vascular endothelial growth factor-B and retinal vascular development in the mouse. AB - PURPOSE: Vascular endothelial growth factor-A (VEGF-A) is crucial to retinal vascular growth, both normal and pathological. VEGF-B, recently characterized, is reported to be expressed in retinal tissues, but the importance of VEGF-B to retinal vascular development remained unknown. The aim of this study was to analyse retinal vascular growth in the Vegfb-/- knockout mouse. METHODS: Retinal vascular growth was measured in Vegfb-/- knockout mice raised under normal conditions, and Vegfb-/- knockout mice with an oxygen-induced proliferative retinopathy. Wild type Vegfb+/+ mice served as controls. Vessels were perfused with ink and retinal flatmounts secondarily labelled with FITC-lectin (BS-1, Griffonia simplicifolia). Area and diameter of retinal growth and retinal vascular growth were recorded over days 0-20, and capillary density and mean diameter recorded from day 17 pups. RESULTS: A variety of techniques confirmed that Vegfb+/+ mice expressed VEGF-B and that VEGF-B expression was absent in Vegfb-/- mice. Vegfb-/- mice raised in room air showed no significant differences from Vegfb+/+ controls. No differences were found in oxygen-induced retinopathy between Vegfb-/- and Vegfb+/+ pups in either the extent of the initial oxygen induced ablation, or in the regrowth of retinal vessels or vitreal (neovascular) sprouts; vitreal sprouts are important markers of the abnormal proliferative response, and are maximally expressed on day 17 in this model of oxygen-induced retinopathy. CONCLUSIONS: These results indicate that a lack of VEGF-B does not significantly affect development of the retinal vasculature under normal conditions, nor does it appear to affect the proliferative retinal responses seen in oxygen-induced retinopathy. PMID- 12580899 TI - Effects of intracameral lidocaine on ocular tissues. AB - PURPOSE: To investigate the ultrastructural changes in iris and corneal tissue induced by intracameral 1% lidocaine infusion applied during lens extraction in a rabbit model. METHODS: The study was conducted using New Zealand rabbits. Eight rabbits received 0.2 mL 1% lidocaine hydrochloride intracamerally and lens extraction was performed, keeping the posterior capsule intact. After lens extraction, cornea and iris tissue samples were obtained for electron microscopy. Eight eyes were included as a control group. RESULTS: Electron microscopy revealed morphological abnor-malities in both cornea and iris of the lidocaine injected eyes, different from the control group. Cytoplasmic vacuolization, phagosomes and residual bodies were observed in epithelial cells. Corneal fibroblasts contained fluid-filled vacuoles, which could be due to the influx of water into the cells as a result of corneal endothelial damage. Mitochondrial swelling and residual bodies were also seen in the cytoplasm of fibroblasts. Blood vessels in the iris contained fluid material composed of fibrin and proteinaceous material and many vacuoles showed vascular endothelial injury. CONCLUSION: Even a short period of exposure of intra-cameral lidocaine to the ocular tissues can induce histo-logical changes that may result in functional defects. PMID- 12580900 TI - Pathogenesis of orbital cholesterol granuloma. AB - Two cases are presented of orbital cholesterol granuloma associated with gradual proptosis arising in men aged 35 and 41 years. Computed tomography demonstrated osteo-lytic masses in the frontal bone at the lacrimal fossa. Curettage revealed a characteristic histology of foreign body reactions surrounding cholesterol clefts. In both cases abnormal bone, more consistent with fibrous dysplasia than reactive change, was found at the periphery. The finding of abnormal bone associated with orbital cholesterol granulomas suggests that a pre-existing bone anomaly may be present in a subset of these cases. PMID- 12580901 TI - Assessing the cornea by in vivo confocal microscopy. PMID- 12580903 TI - Learning from discrepancies: CD14 polymorphisms, atopy and the endotoxin switch. PMID- 12580904 TI - Alcoholic drink consumption: a role in the development of allergic disease? PMID- 12580905 TI - B lymphocytes in allergic alveolitis. PMID- 12580906 TI - Uneasy breather: the implications of dust mite allergens. PMID- 12580907 TI - Evaluation of the CD14 C-159 T polymorphism in the German Multicenter Allergy Study cohort. AB - BACKGROUND: Multiple genetic studies have shown linkage of atopy-related phenotypes to chromosome 5q31. In this region several candidate genes for atopy are localized such as the Th2 cytokines IL-4, IL-5 and IL-13, but also CD14, a receptor for LPS. Recently, a functional CD14 promoter polymorphism was related to total and specific IgE responsiveness. OBJECTIVE: The aim of our study was to evaluate the role of this single nucleotide polymorphism (SNP) in a large German birth cohort. METHODS: Atopy-related phenotypes were longitudinally carefully evaluated in over 800 children from birth to the age of 10 years. Yearly visits included standardized interviews, physical examinations and determination of total and specific IgE antibodies. Pulmonary function tests and histamine provocations were performed at the age of seven. Eight-hundred and seventy-two children of the Multicenter Allergy Study (MAS) cohort were genotyped using melting curve and restriction digest analyses. RESULTS: CD14-159 allele frequencies were consistent with previous reports, however, no association of the SNP with asthma, atopic dermatitis, allergic rhinitis, total or specific IgE levels could be observed. CONCLUSION: The CD14-159 SNP might not play a major role in the development of atopy in German children. PMID- 12580909 TI - Familial and environmental determinants for wheezing and asthma in a case-control study of school children in Palestine. AB - BACKGROUND: Our prevalence study on Palestinian school children aged 6-12 years showed lower rates for asthma and asthma symptoms than economically developed and industrialized countries. Reasons for such differences are largely unknown, and could possibly be related to different environmental and lifestyle factors. OBJECTIVE: To investigate familial, early life exposures and indoor environmental determinants for asthma in children in Palestine. METHODS: From the population of our previous study, a group of 273 children with wheeze in the past 12 months (of whom 99 children had physician-diagnosed asthma) were matched with an equal number of non-wheezing controls. This case-control study involved a parental questionnaire; skin prick testing (SPT) with mixed house dust mites, cat and dog dander, mixed grass, mixed trees pollen, Alternaria tenuis, olive tree and cockroach extracts; and serum for total and specific IgE for the same eight allergens. RESULTS: Paternal asthma and maternal hayfever significantly tripled the risk for their children to have wheezing. Previous diagnoses of bronchial allergy, bronchitis, pneumonia, or whooping cough, and positive SPT for house dust mites and cockroaches were significantly more likely among wheezing and asthmatic children than controls. Specific IgE levels for house dust mites and cat allergens showed significantly higher risk for reported wheezing. After adjustment for several environmental and sociodemographic factors using multivariate logistic regression analysis, paternal asthma, maternal hayfever, damp houses, cat and cockroach SPT positivity proved to be strong predictors for wheezing symptoms. CONCLUSION: Our study confirmed that familial 'atopic' diseases are significant predictors of childhood asthma in Palestinian children. Moreover, indoor environment such as presence of cats and domestic moulds also appear to play a role. Our findings are consistent with studies in Canada, New Zealand, Estonia and Sweden, and show promise to explore further gene-environment interaction in the genesis of asthma. PMID- 12580908 TI - Sibling effect on atopy in children of patients with asthma. AB - BACKGROUND: Multiple population studies have shown the presence of a sibling effect on atopic disease. However, it is unclear if the sibling effect is also of importance in subjects who are genetically at high risk for the development of atopy. OBJECTIVE: To study the presence of a sibling effect on markers of atopy (serum total IgE, specific IgE, skin tests) and asthma (bronchial hyper responsiveness to histamine) in families ascertained through a parent with asthma. METHODS: First-degree offspring in 200 asthma families were studied (n = 541). Mixed effects regression models were used to account for the dependence of the observations within a family, and to adjust for possible confounding variables. RESULTS: Multiple regression analysis showed that having older siblings was inversely related to atopy, defined as >/= 2, >/= 3, >/= 4, or >/= 5 skin tests (P = 0.07-0.009). In addition, family size (number of siblings) had a significant protective effect on the presence of specific IgE to common aeroallergens (P = 0.03). Exposure to cigarette smoke in the first 3 years of life significantly increased the risk of having specific IgE to common aeroallergens (P = 0.04). No sibling effect was detected for serum total IgE or bronchial hyper-responsiveness to histamine. CONCLUSIONS: This study shows a protective sibling effect on the presence and severity of atopy but not on bronchial hyper-responsiveness in children who are genetically at risk. The identification of the sibling effect in high-risk families stresses the need to understand the basis of this effect, in order to design future prevention programmes. PMID- 12580910 TI - The relationship between exhaled nitric oxide and allergic sensitization in a random sample of school children. AB - BACKGROUND: Exhaled nitric oxide (NO) has been proposed as novel a non-invasive marker of airway inflammation. OBJECTIVE: The level of exhaled NO was determined in a random sample of school children (7-12 years old) with the aim of investigating the relationship between exhaled NO and sensitization to common allergens. RESULTS: In the 450 children tested by skin prick tests (SPT), the prevalence of sensitization was 29.5% (overall), 21.9% (sensitization to indoor allergens), and 15.0% (sensitization to outdoor allergens). Regression analysis showed that levels of exhaled nitric oxide were closely associated with various measures of sensitization to aeroallergens. Sensitization to indoor allergens was associated with higher levels of exhaled NO (eNO) than sensitization to outdoor allergens when assessed by IgE but not when assessed by SPT. Children with reported wheeze in the past 12 months had much stronger associations between sensitization and eNO than children without wheeze. CONCLUSION: We conclude that allergic sensitization is strongly associated with increased levels of exhaled NO, especially in children with wheeze. PMID- 12580911 TI - The relationship of alcohol consumption to total immunoglobulin E and the development of immunoglobulin E sensitization: the Copenhagen Allergy Study. AB - BACKGROUND: Several studies in patient populations have reported a positive association between alcohol consumption and serum total IgE. Furthermore, we have previously reported a positive association between alcohol consumption and the prevalence of skin prick test (SPT positivity) to inhalant allergens in a population-based cross-sectional study. OBJECTIVE: To investigate the relationship of alcohol consumption to levels of serum total IgE and the development of IgE sensitization to inhalant allergens. METHODS: In 1990, self reported consumption of alcohol, serum total IgE, SPT positivity and specific IgE positivity to inhalant allergens were assessed in 1112 subjects, aged 15-69 years, participating in a population-based cross-sectional study in Copenhagen, Denmark. In 1998, they were invited to a follow-up and 734 were re-examined (participation rate 69.0%). Adjustment for potential confounders was performed by using multivariable regression analyses. RESULTS: In non-atopic (specific IgE negative) subjects there was a positive association between alcohol consumption and the concentration of total IgE (P = 0.001). During the follow-up period, 45 and 33 subjects developed SPT positivity and specific IgE positivity, respectively. There was no significant association between alcohol consumption and the development of SPT positivity or specific IgE positivity. However, the risk of developing SPT positivity tended to increase with increasing consumption of alcohol (P = 0.055). CONCLUSIONS: This epidemiological study confirms that alcohol consumption has an influence on levels of serum total IgE. A significant association between alcohol consumption and the development of IgE sensitization was not established. However, there seems to be a lack epidemiological data on this issue. PMID- 12580912 TI - Association of alcohol consumption with total serum immunoglobulin E levels and allergic sensitization in an adult population-based survey. AB - BACKGROUND: Chronic alcoholism is associated with increased total serum IgE levels. OBJECTIVE: The study aimed to investigate the relationship between alcohol intake and both total serum IgE levels and allergic sensitization in a general adult population. MATERIALS AND METHODS: A total of 720 subjects was randomly selected (stratified by age) from the population older than 18 years of A-Estrada (Spain) and invited to participate in the study. From 697 eligible subjects, 469 (67%, median age 54 years, range 18 to 92 years, 44% males, 75% of cases from a rural environment) agreed to participate. A battery of 13 skin prick tests to common aeroallergens was performed in all subjects. Cases with at least one positive test (n = 121, 26%) were considered to have allergic sensitization. The most frequent sensitisers were mites and pollens (24% and 10% of subjects, respectively). Total serum IgE was measured in 465 subjects (99%). Alcohol consumption was registered as the number of standard (approximately 10 g) drinking units habitually consumed per week. A total of 244 subjects (52%) were alcohol consumers (median intake, 14 units/week, range 1 to 147 units/week). Abstainers (n = 225, 48%) constituted the reference category. RESULTS: Alcohol consumption of more than 14 units/week was associated with an increase in serum IgE levels after adjusting for age, gender, allergic sensitization and smoking (P = 0.02). Alcohol consumption was not significantly associated with either overall allergic sensitization or mite sensitization after adjusting for age, gender and smoking. However, alcohol consumption of more than 14 units/week was associated with an increased prevalence of pollen sensitization (adjusted OR 3.15, 95% CI 1.19 to 8.34, P = 0.02). CONCLUSION: Alcohol consumption above a certain threshold is associated with an increase in total serum IgE levels. Alcohol consumption may also be associated with an increased prevalence of pollen sensitization. PMID- 12580913 TI - Long-lasting effect of sublingual immunotherapy in children with asthma due to house dust mite: a 10-year prospective study. AB - BACKGROUND: Subcutaneous immunotherapy for respiratory allergy has shown a long lasting efficacy after its discontinuation, whereas this evidence is still lacking for sublingual immunotherapy, despite the fact that it is widely used. OBJECTIVE: We aimed to evaluate whether a long-lasting effect of SLIT occurs, in a prospective parallel group controlled study. METHODS: Sixty children (mean age 8.5 years) suffering from allergic asthma/rhinitis due to mites were subdivided into two matched groups: 35 underwent a 4- to 5-year course of SLIT with standardized extract and 25 received only drug therapy. The patients were evaluated at three time points (baseline, end of SLIT and 4 to 5 years after SLIT discontinuation) regarding presence of asthma, use of anti-asthma drugs, skin prick tests and specific IgE. RESULTS: We found that in the SLIT group there was a significant difference vs. baseline for the presence of asthma (P 100 mg/kg/day) compared to the 11 patients with secondary hyperparathyroidism. However, variation of all parameters was great with considerable overlap. Very high S-PTH levels > or = 42 pmol/l were observed in those who later developed tertiary hyperparathyroidism. CONCLUSIONS: Prolonged very high dose oral Pi treatment is a major risk factor for the development of tertiary hyperparathyroidism in X-linked hypophosphatemic rickets. PMID- 12580930 TI - Diagnosis of growth hormone deficiency after pituitary surgery: the combined acipimox/GH-releasing hormone test. AB - OBJECTIVE: Reduction of plasma free fatty acids leads to enhanced GH response after stimulation by GH-releasing hormone (GHRH). We studied the clinical usefulness of combined administration of acipimox and GHRH for the diagnosis of GH deficiency. DESIGN: We evaluated 35 patients [mean age 53.0 years; mean body mass index (BMI) 26.7 kg/m2] after pituitary surgery. We compared GH responses after acipimox and GHRH with the GH response during an insulin tolerance test (ITT) and, in a subgroup of 12 patients, with the GHRH/arginine test. The acipimox/GHRH test was additionally performed in 21 control subjects (mean age 53.8 years; mean BMI 24.7 kg/m2). RESULTS: In the patients, the mean (+/- SEM) peak GH was almost four-fold higher after acipimox/GHRH (6.94 +/- 1.07 microg/l, range 0.46-23.1; P < 0.001) and after GHRH/arginine (8.32 +/- 1.23 microg/l, range 1.1-49.2; P < 0.001) than after ITT (1.84 +/- 0.46 microg/l, range 0.01 11.9). According to the ITT, 29 patients were severely GH deficient (peak GH < 3.0 microg/l). Peak GH levels after acipimox/GHRH in controls ranged from 7.5 to 78.4 microg/l (mean 29.3 +/- 3.5). GH peak values during the acipimox/GHRH test were significantly correlated with values from the ITT (r = 0.63, P < 0.01) and GHRH/arginine test (r = 0.87, P < 0.001). Areas under the curve were also correlated. According to generally accepted cut-off peak GH levels for the ITT and GHRH/arginine test, a GH peak exceeding 11.2 micro g/l excludes severe GH deficiency after acipimox/GHRH. Our control data indicate that the cut-off level is lower at older age. CONCLUSIONS: The acipimox/GHRH test leads to GH responses similar to those of the GHRH/arginine test, and to higher peak GH values if compared with the ITT. The acipimox/GHRH test is a potential additional tool to detect GH deficiency in patients with pituitary disease, in particular in patients with a perturbation of fatty acid metabolism. PMID- 12580932 TI - Reversal of acromegalic cardiomyopathy in young but not in middle-aged patients after 12 months of treatment with the depot long-acting somatostatin analogue octreotide. AB - BACKGROUND: Cardiovascular disease is the most frequent cause of death of patients with acromegaly. AIM: To investigate whether young patients with a presumed short disease duration are more likely to reverse the acromegalic cardiomyopathy than older patients with longer disease duration. DESIGN: An open prospective design. PATIENTS: Ten young (aged < 40 years), and 12 middle-aged (41 59 years) patients with active acromegaly well controlled after 12 months of treatment with the depot formulation of octreotide (OCT-LAR); 22 sex- and age matched healthy subjects as controls. METHODS: Left ventricular (LV) mass (LVM) by echocardiography and performance by equilibrium radionuclide angiography were measured before and after 12 months of OCT-LAR treatment. RESULTS: At study entry, none of the controls and 14 patients (63.4%) of whom six were young (chi2 = 17.7; P < 0.0001) had LV hypertrophy (LVH); none of the controls and four patients of whom one was young had insufficient LV ejection fraction (LVEF) at rest (< 50%); one control and 13 patients (59.1%) of whom five were young (chi2 = 12.7; P < 0.0001) had inadequate LVEF at peak exercise (deltaLVEF; < 5% increase of baseline). After 12 months, no change in haemodynamic and diastolic parameters was observed in both groups, except for a significant decrease in heart rate at peak exercise in young patients (P < 0.0001). The LVM index decreased significantly in both young (124.4 +/- 5.8 vs. 103.4 +/- 3.9 g/m2; P = 0.01) and middle-aged patients (140.9 +/- 7.9 vs. 117.8 +/- 6.6 g/m2; P = 0.03). LVH disappeared in 10 of 14 patients (71.4%): all six young and four of eight middle aged patients (50%). LVEF at rest and at peak exercise increased significantly in both groups but deltaLVEF increased significantly only in young patients (1.5 +/- 2.9 vs. 13.7 +/- 5.2%, P = 0.04); it normalized in nine of 13 patients (69.2%), four of five young (80%) and four of eight middle-aged patients (50%). Exercise capacity (82.5 +/- 5.3 vs. 110.0 +/- 5.5 W, P = 0.005) and duration (7.3 +/- 0.3 vs. 9.9 +/- 0.4 min, P = 0.0003) also increased only in young patients. CONCLUSIONS: The acromegalic cardiomyopathy is reversed in most young patients with short disease duration and achieving disease control after OCT-LAR treatment for 12 months, indicating that early diagnosis and effective treatment are essential. PMID- 12580933 TI - Endocrine profiles and semen quality in spinal cord injured men. AB - PURPOSE: To evaluate the hypothalamic-pituitary-testis (HPT) axis, endocrine profiles and semen quality in men with spinal cord injury (SCI). MATERIALS AND METHODS: Fifty-five men with SCI were studied. Serum levels of FSH, LH, testosterone, oestradiol and prolactin (PRL) were determined; the LH-releasing hormone (LHRH) stimulation test and a semen analysis were performed, and testicular volumes were measured. Thirty-six age-matched healthy male volunteers and 34 noninjured infertile men served as controls. RESULTS: Eight SCI subjects had low basal LH, four had low basal FSH, and 16 had decreased basal serum levels of LH and FSH. Of subjects with lower serum levels of gonadotrophins (LH and/or FSH), nine had low serum testosterone and seven had hyperprolactinaemia. Serum levels of oestradiol were similar for all groups. There were increased LH and FSH responses to LHRH in SCI subjects compared to normal controls, but this difference was only statistically significant in SCI subjects with lower than normal serum levels of LH and/or FSH. There was no significant difference in testis volume between SCI subjects and controls. The mean semen volume in SCI subjects was lower than from controls, but the difference was not statistically significant. Sperm motility and percent normal sperm morphology were lower in SCI compared to normal controls but not to infertile control subjects. In total, 51% and 86% of SCI subjects had at least one hormonal or axis abnormality, respectively. CONCLUSION: We conclude that hypogonadotropism in SCI subjects is likely to be secondary to altered neural or hormonal pathways between the hypothalamus and the pituitary gland, and that these endocrine abnormalities may be the mechanisms contributing to impairment of semen quality. PMID- 12580934 TI - Hyperprolactinaemia in hypothyroidism: clinical significance and impact of TSH normalization. AB - OBJECTIVES: Menstrual irregularities in hypothyroidism have been reported to occur less frequently than previously described. We therefore studied the influence of serum PRL in patients with newly diagnosed subclinical and overt hypothyroidism and in hyperprolactinaemic patients treated with T4 to distinguish the impact of hypothyroidism from that of confounding drugs on hyperprolactinaemia and menstrual irregularities. PATIENTS AND METHODS: PRL was determined in 1003 consecutive hypothyroid patients (TSH > 4.0 mU/l) at referral, and after TSH normalization in 84 (8%) initially hyperprolactinaemic (female, > 480 mU/l; male, > 432 mU/l) subjects. Medical history (psychotropic drugs and oestrogens) and menstrual patterns were assessed at referral and after 8 +/- 5 (mean +/- SD) months of T4 therapy. Pituitary magnetic resonance imaging (MRI) was offered to patients with persistently elevated PRL. RESULTS: Menstrual disturbancies (oligomenorrhoea/secondary amenorrhoea, O/A) were not more common (P = NS) in hyper- than in normoprolactinaemic women (26% and 16%, respectively). We observed no galactorrhoea and no correlation between PRL and TSH or O/A except in pregnant or lactating women (N = 11). Oestrogens or antidepressants (including selective serotonin reuptake inhibitors) did not cause hyperprolactinaemia but antipsychotic drugs did. PRL decreased with T4 therapy (P < 0.01) in patients not using confounding drugs (from 720 +/- 288 to 360 +/- 192 mU/l) but menstrual irregularities persisted. PRL remained unchanged in patients receiving antipsychotic treatment. PRL was also unchanged in patients with pituitary abnormalities (seven micro-, one macroadenoma). CONCLUSIONS: Hyperprolactinaemia was not an important feature in patients with newly diagnosed hypothyroidism. Neuroleptic drugs may cause persisting hyperprolactinaemia after TSH normalization. In addition, menstrual disturbancies do not relate to hyperprolactinaemia in hypothyroidism. PMID- 12580935 TI - Facial disfigurement: is it in the eye of the beholder? A study in patients with Graves' ophthalmopathy. AB - OBJECTIVES: The importance of facial disfigurement in many diseases necessitates a reliable and valid measure of disfigurement severity for clinical studies. The hypothesis is that a universal concept of disfigurement exists and can be measured in a reliable way. The objectives of this study were to investigate if persons, in particular patients and physicians, can agree on facial disfigurement severity; and to determine the relative contribution of predefined clinical characteristics of patients with Graves' ophthalmopathy (GO) to the overall rating of facial disfigurement severity. DESIGN: A panel study was carried out in four different panels, each consisting of four members. PATIENTS: We randomly selected 100 slide pairs of GO patients from four available study populations, involving mild, moderate and severe GO patients (mean age 49 years, 76% female) who were treated with either radiotherapy, sham-irradiation, prednisone or orbital decompression. MEASUREMENTS: All panel members individually scored the disfigurement severity of 100 GO patients shown on standardized slides on a Visual Analog Scale. In total, 1600 ratings were collected. We calculated within- and between-panel agreement of disfigurement severity and identified determinants of disfigurement. RESULTS: Agreement within a panel varied from 0.65 to 0.79 and was highest within the panel of ophthalmologists. Between-panel agreement was 0.67 and was highest between ophthalmologists and laypersons. Compared with the global average, patients overrated and endocrinologists underrated disfigurement severity. Female panellists rated the patients, on average, more disfigured than male panellists. Important determinants of disfigurement were eyelid retraction, severe eyelid swelling and proptosis. Their relative importance was consistent across panels and in contrast to current measures of GO severity. CONCLUSION: Facial disfigurement severity can be measured in a reliable way using panels of panellists. Except for some systematic differences between panellists, facial disfigurement does not seem to be in the eye of the beholder. PMID- 12580936 TI - Two Italian kindreds with familial hypocalciuric hypercalcaemia caused by loss-of function mutations in the calcium-sensing receptor (CaR) gene: functional characterization of a novel CaR missense mutation. AB - OBJECTIVES: Description of two unrelated Italian kindreds with familial hypocalciuric hypercalcaemia (FHH), an autosomal dominant disease mostly caused by heterozygous inactivating mutations of the Ca2+ sensing receptor (CaR). PATIENTS AND DESIGN: We studied 11 members of the two families. Genomic DNA was isolated from peripheral blood leucocytes in all family members and in 50 unrelated Italian controls. Total serum and ionized calcium, PTH, creatinine, phosphate, magnesium, and urinary calcium clearance to creatinine clearance ratio were measured. Direct sequencing of the entire coding region of the CaR was performed in the probands. Functional studies were performed in COS-7 cells transiently expressing the mutated CaR. RESULTS: In the proband of family A direct sequencing revealed a novel heterozygous Y218C missense mutation in exon 4. The same mutation was identified in the affected but not in the unaffected family members or in any of the 50 unrelated Italian controls. Transient expression of the Y218C CaR in COS-7 cells revealed a blunted Ca2+-evoked accumulation of inositol trisphosphates, indicating that the Y218C is a loss-of function mutation. Cotransfection experiments showed that the mutant receptor had no impact on the function of the wild-type receptor, suggesting that a reduced expression of the normal CaR, rather than a dominant-negative effect, accounted for the functional impairment. In the proband of family B an already described heterozygous P55L missense mutation in exon 2 of the CaR gene was found. The same mutation was identified in the affected family members. CONCLUSIONS: We described two familial hypocalciuric hypercalcaemia kindreds with loss-of-function mutations of the Ca2+ receptor gene and identified a novel heterozygous mutation (Y218C) characterized by a blunted response to Ca2+ stimulation compared to the wild-type receptor and no interference with the function of the wild-type Ca2+ receptor. PMID- 12580937 TI - Increased serum reverse triiodothyronine levels at diagnosis of hepatocellular carcinoma in patients with compensated HCV-related liver cirrhosis. AB - OBJECTIVE: The aim of this study was to investigate changes in thyroid hormone metabolism in relation to the development of hepatocellular carcinoma (HCC) in patients with HCV-related liver cirrhosis. MATERIALS AND METHODS: The study group (Group A) comprised 31 patients (25 M, 6 F; median age 62.1 years, range 54.0 81.5 years) affected by HCV-related liver cirrhosis with superimposed HCC. Acute and chronic systemic disease, other than cirrhosis, inducing 'euthyroid sick syndrome' was excluded in all patients. Serum TSH, FT4, FT3, rT3, and thyroxine binding globulin (TBG) levels were retrospectively evaluated in frozen aliquots drawn at the time of tumour diagnosis and every 6 months for 3-7 years before HCC diagnosis. The control group (Group B) comprised 29 patients affected by HCV related liver cirrhosis without HCC, matched for sex, age and grade of liver dysfunction. RESULTS: At the time of HCC diagnosis, all patients in Group A were euthyroid with serum TSH, FT4, FT3 and TBG values not significantly different from those of cirrhotic patients of Group B. However, at diagnosis Group A patients had serum rT3 values that were significantly higher than those in Group B (35.0 ng/dl, range 12.0-162.0 vs. 19.0 ng/dl, range 10.0-51.0; Group A vs. Group B; P < 0.001). Serum rT3 values above the normal range were found in 12 patients in Group A (38.7%) but in only one of the patients from Group B (3.4%) (chi2 10.2; P = 0.001). The serum rT3 levels were not significantly correlated to the Child grade of liver cirrhosis (rho 0.1; P = 0.5). The intrasubject analysis demonstrated that a significant increase in serum rT3 levels occurred at the time of HCC diagnosis but serum FT4, FT3 and TSH values did not change significantly. A receiver operating curve (ROC) demonstrated that a 6-monthly increase in serum rT3 levels of at least +22.5% identified patients with HCC with a diagnostic accuracy of 81.7%. CONCLUSIONS: Our study has demonstrated that development of hepatocellular carcinoma is accompanied by a significant increase in serum rT3 levels in patients with low-grade HCV-related liver cirrhosis who had no other illness causing the 'euthyroid sick syndrome'. PMID- 12580938 TI - Interactions between serum leptin, the insulin-like growth factor-I system, and sex, age, anthropometric and body composition variables in a healthy population randomly selected. AB - OBJECTIVE: Leptin secretion is influenced by many factors and the GH/IGF axis plays an important role in the regulation of body composition, but the physiological interactions between leptin and the IGF-I system remain unknown. In this study we investigated the relationship between leptin, the IGF-I system, and sex, age, anthropometric and body composition variables in a group of healthy adults randomly selected. DESIGN: A cross-sectional study. PATIENTS: The study included 268 subjects, representative of the whole population of the city of L'Hospitalet de Llobregat in sex and age distribution: 134 men aged 41.4 years, range 15-70 years; and 134 women, aged 40.7 years, range 15-70 years. MEASUREMENTS: Body mass index (BMI) was calculated, and body composition was determined by using a bioelectrical impedance analyser. Serum leptin concentrations were determined by using a radioimmunoassay (RIA). Serum total IGF I concentrations, after acid-ethanol extraction, were also measured by RIA. Serum free IGF-I concentrations were determined by an enzymoimmunometric assay. Serum IGFBP3 concentrations were determined by RIA. Plasma basal TSH concentrations were determined by a specific electrochemiluminescence assay. RESULTS: In men the BMI was similar in all decades and waist/hip ratio increased in the last three decades. Fat-free mass decreased by decade. We observed an increase in leptin in the fourth decade with a decrease in IGF-I, free IGF-I and IGFBP3 throughout the decades. Basal TSH showed an increase in the last two decades. In women, BMI, waist/hip ratio and fat mass increased significantly in the last decades. Leptin concentrations increased in the last decades and total IGF-I, free IGF-I and IGFBP3 decreased by decade without changes in basal TSH concentration. In men, there was a positive correlation between leptin and BMI, waist/hip ratio, total body water, fat-free mass and fat mass, and these anthropometric and body composition variables showed a negative correlation with free IGF-I and IGFBP3, without any correlation with total IGF-I. In women, there was a positive correlation between leptin and BMI, waist/hip ratio, total body water, fat-free mass, and fat mass, which showed a negative correlation with total IGF-I and IGFBP3, without any correlation with free IGF-I. In men, total IGF-I was negatively correlated with waist/hip ratio without any correlation with the other variables and free IGF-I was negatively correlated with BMI and waist/hip ratio, and IGFBP3 did not show any correlation. In women, total IGF-I, free IGF-I and IGFBP3 were negatively correlated with BMI, waist/hip ratio and fat mass. The multiple linear regression analysis produced a model that explained 60.5% of leptin variability in men and 40% in women. Notably, only age, BMI, fat mass and waist/hip ratio brought an independent significant contribution to leptin variability. The final model also explained 28.2% and 60.4% of total IGF-I variability and 17.2% and 27.4% of free IGF-I variability in men and women, respectively. Age and leptin contributed to free IGF-I variability in men, and age and fat mass were significantly and independently associated with total IGF-I in women. CONCLUSIONS: In this well-characterized population of controls randomly selected without chronic disease or drug administration and with biochemically confirmed euthyroidism, we found that both men and women had a significant correlation between leptin levels and the IGF-I system, and anthropometric and body composition variables, but that leptin did not regulate the IGF-I system, and that the IGF-I system did not regulate leptin synthesis and secretion. PMID- 12580939 TI - Both fasting-induced leptin reduction and GH increase are blunted in Cushing's syndrome and in simple obesity. AB - BACKGROUND: Simple obesity and Cushing's syndrome (CS) are two clinical models of leptin hypersecretion coupled with GH hyposecretion. Fasting inhibits leptin while stimulating GH secretion in normal human subjects. OBJECTIVES: To clarify the effect of fasting on leptin and GH secretion in obesity and CS. PATIENTS AND PROTOCOL: We studied six women with CS [age 17-66 years; body mass index (BMI) 28.6 kg/m2], seven women with visceral obesity (OB; 20-41 years; BMI 42.9 kg/m2) and seven normal women (NS; 25-31 years; BMI 19.3 kg/m2). The effects of 36-h fasting on 8-h diurnal mean leptin, GH, insulin and glucose concentrations (mLEPTc, mGHc, mINSc and mGLUc) as well as on the IGF/IGFBP system were studied. RESULTS: Before fasting, mLEPTc in OB and in CS were similar and both were higher (P < 0.01) than in NS. OB and CS showed similar mGHc, which were lower (P < 0.05) than in NS. Fasting induced a reduction in mLEPTc that was significant in NS and CS (P < 0.04) but not in OB. The mLEPTc in OB and CS after fasting remained higher (P < 0.05) than in NS. After fasting, OB and CS showed no increase in mGHc, although this clearly increased (P < 0.02) in NS. IGF-I but not IGFBP-3 levels decreased in all groups (P < 0.05). Fasting reduced mINSc and mGLUc while increasing IGFBP-1 in all groups. After fasting, mINSc in OB and CS remained higher (P < 0.03) than in NS. CONCLUSIONS: Short-term fasting has less inhibitory effect on leptin and no stimulatory effect on GH secretion in patients with Cushing's syndrome as well as simple obesity. After fasting, insulin levels in hypercortisolaemic and also in obese patients remained higher than in normal subjects suggesting that hyperinsulinism could play a role in the altered response of leptin and GH to starvation in these conditions. PMID- 12580940 TI - Do growth hormone (GH) serial sampling, insulin-like growth factor-I (IGF-I) or auxological measurements have an advantage over GH stimulation testing in predicting the linear growth response to GH therapy? AB - OBJECTIVE: To compare the relative utility of GH secretion via pharmacological stimulation, overnight serial sampling, IGF-I levels and auxological variables as predictors of change in height standard deviation score (deltaHt SDS) during GH treatment. DESIGN: A multicentre observational study. PATIENTS: Prepubertal children (n = 825) with idiopathic growth failure who were subsequently treated with GH were divided into two groups, based on their maximum GH response to pharmacological stimulation testing: (1) idiopathic GH deficiency (IGHD), defined by a maximum GH response < 10 microg/l (n = 300); and (2) idiopathic short stature (ISS), with a maximum GH response > or = 10 microg/l (n = 525) (GH conversion factor: 3 IU = 1 mg). MEASUREMENTS: Overnight spontaneous GH secretion was measured in all patients. The following characteristics of spontaneous GH secretion were studied: maximum or peak GH, mean peak GH, number of GH peaks, pooled GH, mean GH, and approximate entropy of GH secretion. RESULTS: Although children with IGHD had lower indices of spontaneous GH secretion, there were no differences between IGHD and ISS groups in baseline Ht SDS, growth rate or IGF-I level. The dose and duration of GH therapy were similar. There was no statistically significant difference in the mean (+/- SD) change in Ht SDS (deltaHt SDS) in the two groups (IGHD 1.3 +/- 0.9 and ISS 1.2 +/- 0.8). Measures of spontaneous secretion, such as peak GH, mean of GH peaks, mean area under GH peaks, and mean GH, as well as IGF-I concentrations, were all statistically significantly correlated with deltaHt SDS in IGHD children (P < 0.0001). A significant correlation was also observed for pooled GH (P = 0.002) and approximate entropy (P = 0.01). Children with the most severe ISS (Ht SDS < 3.33) demonstrated a more disorganized pattern of GH secretion compared to children who were not as short (Ht SDS -2.33 to -1.64), as indicated by a higher approximate entropy (0.673 +/- 0.193 vs. 0.607 +/- 0.161, P < 0.004). This increased disorder in GH secretion was accompanied by lower IGF-I levels (104 +/- 99 microg/l vs. 137 +/- 74 microg/l, P < 0.001), even though pooled GH concentrations were indistinguishable between the two groups (2.2 +/- 1.3 microg/l vs. 2.0 +/- 1.0 microg/l). Children with IGHD demonstrated lower approximate entropy than did those with ISS (0.551 +/- 0.235 vs. 0.631 +/- 0.182, P < 0.0001). Duration of GH treatment, height deficit and genetic potential (midparental Ht SDS) were the most important variables influencing deltaHt SDS in children receiving GH therapy. Maximum stimulated GH, IGF-I and indices of spontaneous GH secretion also correlated with deltaHt SDS, but their relative importance varied among diagnostic groups. CONCLUSIONS: Patients with GH deficiency demonstrate a reduced capacity for GH secretion, while those with idiopathic short stature exhibit a more disorderly and less functional secretory pattern. Although effective in predicting a response to GH treatment in patients with severe GH deficiency, overnight serial sampling is less practical than other methods currently available. In addition, serial sampling was less useful as a predictor of growth response to exogenous GH in patients with idiopathic short stature. PMID- 12580941 TI - The insulin-like growth factor-I generation test: peripheral responsiveness to growth hormone is not decreased with ageing. AB - OBJECTIVE: Ageing is accompanied by a reduction in GH secretion, and a decrease in circulating IGF-I. Few data are available on whether the responsiveness of IGF I to GH stimulation changes with age. SUBJECTS AND METHODS: Therefore we carried out multiple IGF-I generation tests in 26 healthy volunteers (16 male) of normal body mass index (BMI); nine aged 20-40 years, six aged 41-60 years, and 11 aged > 61 years. Each subject received three single doses of GH: 0.8, 2.0 and 21 IU in random order at least 4 weeks apart. Serum samples were taken 0, 18, 24, 48, 72 and 120 h following each dose of GH. RESULTS: Basal serum levels of IGF-I (P < 0.0001) and IGFBP-3 (P < 0.01) declined with age, but serum acid-labile subunit (ALS) levels did not (P = 0.2). Peak IGF-I levels (P < 0.01 for 0.8 IU and P < 0.05 for the 2 IU dose) and area under curve (AUC) IGF-I (P < 0.01 for the 0.8 IU and 2.0 IU doses of GH and P < 0.05 for the 21 IU dose) after GH administration continued to demonstrate a significant trend towards lower values with increasing age. However, the increment in IGF-I, IGFBP-3 and ALS in response to GH did not decline with age. Indeed, the increment in IGF-I after 2 IU of GH, judged by the increase from basal to peak levels, increased with advancing age (P = 0.05), and a positive relationship was seen between the increment in the area under the IGF I curve following the 21 IU dose of GH and age (P < 0.02). CONCLUSION: These data illustrate that although activity of the GH/IGF-I axis declines with age, peripheral responsiveness to GH is not attenuated. This suggests that a decrease in GH responsiveness does not contribute to the age-related fall in circulating GH-dependent peptides. Thus, for those embarking on trials of GH therapy or GH secretagogues in the elderly, the capacity to generate IGF-I will not limit potential efficacy. Furthermore, the dose of GH replacement required for patients with organic GH deficiency is likely to be lower in the elderly compared with young adults. PMID- 12580942 TI - Different effectiveness of cyproterone acetate doses in treatment of acne. PMID- 12580943 TI - Molecular analysis of the 21-hydroxylase gene. PMID- 12580944 TI - In the belly of the beast: subversion of macrophage proinflammatory signalling cascades during Toxoplasma gondii infection. AB - Macrophages (MO) are used as the intracellular niche by several bacterial and protozoan microorganisms. Such microbial pathogens adopt diverse strategies to avoid MO microbicidal effects. Recent insights into the Toxoplasma gondii-MO interaction reveal novel ways that intracellular parasites subvert MO function. In contrast to some microbial pathogens, Toxoplasma infection is not silent but induces rapid activation of transcription factors such as STAT-1 and NFkappaB. However, the parasite blocks nuclear translocation of both factors, and MO cannot produce IL-12 or TNF-alpha when subsequently triggered with lipopolysaccharide. The nuclear import blockade is lifted 24 h after infection, but cells remain actively suppressed in TNF-alpha production. Nevertheless, IL-12 synthesis is initiated at this later time point. Toxoplasma gondii-induced production of this cytokine occurs through both MyD88- and CCR5-dependent pathways. The balance of cytokine subversion and stimulation during infection probably results from the parasite's need to simultaneously avoid immune elimination and trigger immunity to prevent host death. PMID- 12580945 TI - Verotoxin 1 binding to intestinal crypt epithelial cells results in localization to lysosomes and abrogation of toxicity. AB - Verotoxins (VTs) are important virulence factors of enterohaemorrhagic Escherichia coli (EHEC), a group of bacteria associated with severe disease sequelae in humans. The potent cytotoxic activity of VTs is important in pathogenicity, resulting in the death of cells expressing receptor Gb3 (globotriaosylceramide). EHEC, particularly serotype O157:H7, frequently colonize reservoir hosts (such as cattle) in the absence of disease, however, the basis to avirulence in this host has been unclear. The objective of this study was assessment of interaction between VT and intestinal epithelium, which represents the major interface between the host and enteric organisms. Bovine intestinal epithelial cells expressed Gb3 in vitro in primary cell cultures, localizing specifically to proliferating crypt cells in corroboration with in situ immunohistological observations on intestinal mucosa. Expression of receptor by these cells contrasts with the absence of Gb3 on human intestinal epithelium in vivo. Despite receptor expression, VT exhibited no cytotoxic activity against bovine epithelial cells. Sub-cellular localization of VT indicated that this toxin was excluded from endoplasmic reticulum but localized to lysosomes, corresponding with abrogation of cytotoxicity. VT intracellular trafficking was unaffected by treatment of primary cell cultures with methyl-beta-cyclodextrin, indicating that Gb3 in these cells is not associated with lipid rafts but is randomly distributed in the membrane. The combination of Gb3 isoform, membrane distribution and VT trafficking correlate with observations of other receptor positive cells that resist verocytotoxicity. These studies demonstrate that intestinal epithelium is an important determinant in VT interaction with major implications for the differential consequences of EHEC infection in reservoir hosts and humans. PMID- 12580946 TI - Neisserial PorB is translocated to the mitochondria of HeLa cells infected with Neisseria meningitidis and protects cells from apoptosis. AB - We have previously shown that purified meningococcal porin PorB associates with mitochondria and prevents apoptosis of B cells, Jurkat cells and HeLa cells (Massari et al., 2000, Proc Natl Acad Sci USA 97: 9070-9075). This work examines if intact meningococci have a similar effect as purified porins. It was first determined that intact live meningococci do not induce apoptosis of HeLa cells and do not perturb mitochondrial physiology. This latter consideration is important as Neisserial porins affect the susceptibility of cells to apoptosis by preventing mitochondrial depolarization and cytochrome c release, events involved in the apoptosis cascade. Purified PorB or PorB from live bacteria were found to translocate into and interact with mitochondria. It was then determined whether treatment of HeLa cells with meningococci could prevent staurosporine-mediated apoptosis due to an effect of PorB on the mitochondrial parameters. Incubation of HeLa cells with live meningococci prevented staurosporine-induced apoptosis, as ascertained by measurements of mitochondrial potential, translocation of mitochondrial cytochrome c to the cytosol, caspases activation, and nuclear DNA degradation. These data are consistent with our previous findings that purified PorB associates with mitochondria and prevents apoptosis, and demonstrates that the mechanism by which whole meningococci protects cells from apoptosis is a result of direct interaction of neisserial porin with mitochondria. PMID- 12580947 TI - Interaction of Actinobacillus actinomycetemcomitans outer membrane vesicles with HL60 cells does not require leukotoxin. AB - Outer membrane derived vesicles (MVs) secreted by Actinobacillus actinomycetemcomitans JP2 contain a membranolytic leukotoxin and are toxic to human HL60 cells. To determine how MVs interact with human target cells, HL60 cells were incubated with vesicles, reacted with anti-vesicle antibodies and a FITC-labelled reporter, and visualized by confocal scanning laser microscopy. Target cells rapidly became reactive with anti-vesicle antibodies upon exposure to vesicles. Confocal microscopy showed that labelling occurred primarily in the cytoplasmic membrane and that very little internal fluorescence was observed. The cytoplasmic membrane of HL60 cells was also strongly labelled after exposure to MVs that contained the fluorescent phospholipid, SP-DiOC18. In contrast, incubation of cells with free SP-DiOC18 resulted primarily in the labelling of internal structures of HL60 cells. These results suggest that A. actinomycetemcomitans MVs associate with, or are incorporated into the cytoplasmic membrane of HL60 cells. The leukotoxin is a membranolytic cytotoxin and cells exposed to MVs were lysed by vesicle-associated toxin in a time and dose-dependent manner. However, cells became reactive with anti-vesicle antibodies when MVs were added in the presence of inhibitors of leukotoxin mediated lysis or when sublytic doses of MVs were analysed. In addition, MVs produced by an isogenic leukotoxin-deficient strain of A. actinomycetemcomitans JP2 were non-toxic but rapidly interacted with HL60 cells. These results suggest that A. actinomycetemcomitans MVs can deliver leukotoxin to HL60 cells but that the association of vesicles with the cytoplasmic membrane occurs independently of the leukotoxin polypeptide. PMID- 12580948 TI - Bordetella type III secretion induces caspase 1-independent necrosis. AB - The Bordetella bronchiseptica type III (TIII) secretion system induces cytotoxicity in infected macrophages and epithelial cells. In this report we characterize the cell death phenotype and compare it to the TIII-dependent cytotoxicity induced by Yersinia enterocolitica and Shigella flexneri. Bordetella bronchiseptica strain RB58 was able to induce cell death in J774A.1 macrophages with the same efficiency as Shigella and Yersinia, but only B. bronchiseptica was able to kill epithelial cells in a TIII-dependent manner. Primary macrophages from caspase 1-/- mice were susceptible to RB58-mediated killing, suggesting that unlike Shigella and Salmonella, caspase 1 does not mediate cell death. RB58 induced cytotoxicity was not inhibited by addition of the pan-caspase inhibitor zVAD, and Western blot analyses of RB58-infected HeLa cells indicated that neither caspase 3 nor 7 was cleaved and PARP remained in its full-length active form. Morphologically the RB58-infected HeLa cells resembled necrotic rather than apoptotic cells, exhibiting cytoplasmic swelling and extensive membrane blebbing in the absence of nuclear changes. The addition of exogenous glycine, which has been shown to prevent necrotic cell death by blocking non-specific ion fluxes across the plasma membrane, blocked RB58-induced cytotoxicity. Addition of cyclosporin A which prevents the opening of the mitochondrial permeability pore, had no effect on RB58-infected cells. We conclude that the B. bronchiseptica TIII secretion system induces a mode of cell death consistent with necrosis that is distinct from that of Yersinia and Shigella. PMID- 12580949 TI - May-Hegglin anomaly. PMID- 12580950 TI - Thrombocytopenia and abdominal mass. PMID- 12580952 TI - Human herpesvirus 6: relevance of infection in the immunocompromised host. PMID- 12580951 TI - Pyrimidine 5' nucleotidase deficiency. PMID- 12580953 TI - A nitric oxide synthase inhibitor, N(G)-nitro-l-arginine-methyl-ester, exerts potent antiangiogenic effects on plasmacytoma in a newly established multiple myeloma severe combined immunodeficient mouse model. AB - Angiogenesis is one of critical factors in sustaining the growth, invasion and metastasis of certain solid tumours and haematological malignancies such as multiple myeloma (MM). In the present study, we examined the anticancer potential of an inhibitor of nitric oxide synthase (NOS), NG-nitro-l-arginine methyl ester (L-NAME), in a novel severe combined immunodeficient mouse model (KHM mouse) that harbours the highly sanguineous plasmacytoma cell line KHM-4, derived from a patient with highly chemoresistant MM. KHM mice were intraperitoneally administered with either L-NAME, doxorubicin, melphalan, or paclitaxel. A significant reduction in tumour sizes was noted in L-NAME-administered KHM mice while no significant reduction was observed in melphalan- or doxorubicin administered mice. A profound decrease in angiogenesis was observed in tumour tissues from L-NAME- and paclitaxel-administered KHM mice. A marked decrease in human vascular endothelial cell growth factor (VEGF) levels was identified in tumour tissues from L-NAME-administered KHM mice, strongly suggesting that L-NAME suppressed VEGF production by tumour cells through its inhibition of NOS in tumour cells, resulting in reduced neovasculization in mice leading to the regression of tumour sizes. The present data represent the first observations that certain NOS inhibitors potentially serve as experimental agents for the treatment of chemoresistant MM and plasmacytoma. PMID- 12580954 TI - Peripheral blood stem cell contamination evaluated by a highly sensitive molecular method fails to predict outcome of autotransplanted multiple myeloma patients. AB - To evaluate the clinical impact of minimal residual disease in multiple myeloma, apheretic products from 51 autotransplanted patients were tested by fluorescent (GeneScan) polymerase chain reaction (PCR). Sixty-nine per cent of harvests were contaminated when evaluated for IgH rearrangement. Forty-six patients responded to transplant, with 52.9% achieving complete response (CR). The clinical response of patients was significantly influenced by the number of re-infused CD34+ cells. Positive PCR results of re-infused harvests were not significantly related to patient outcome. Median overall survival (OS) was 33 months, and a significant advantage for patients transplanted by 12 months from diagnosis was observed. Moreover, OS was longer for patients receiving PCR-negative stem cells, with 72% of patients surviving to 70 months in the group receiving PCR-negative harvests vs 48% in the group transplanted with contaminated precursors (not statistically significant). Ex vivo purging caused a reduction of contamination of up to 3 logs; nevertheless, 80% of purged harvests remained PCR-positive and the purging procedure did not alter response or survival rates. Thus, the failure of a predictive role for this highly sensitive molecular method could be explained by the assumption that in vivo persisting malignant cells are the true source of relapse in MM. PMID- 12580955 TI - Mobilization of CD34+ cells in elderly patients (>/= 70 years) with multiple myeloma: influence of age, prior therapy, platelet count and mobilization regimen. AB - The mobilization of peripheral blood stem cells was studied in 984 multiple myeloma patients, including 106 patients aged >/= 70 years. Increasing age correlated inversely with CD34+ yield (P < 0.0001), but also with >/= 12 months of prior standard chemotherapy (P = 0.0001), < 200 x 10(9)/l platelets (P = 0.0006) premobilization and mobilization with growth factors only (P = 0.0001). After controlling for these age covariates, multivariate analysis identified /= 200 x 10(9)/l premobilization as favourable variables (both P < 0.0001), while increasing patient age remained an unfavourable factor (P = 0.0009). With both favourable variables, 85% of elderly patients collected >/= 4 x 10(6)/kg CD34+ cells in a median of one collection. The effect of age was incremental with no age threshold showing acceleration in the decline of CD34+ yield. Chemotherapy significantly increased CD34+ yield compared with growth factors only. However, the subgroup of patients with > 12 months prior therapy and premobilization platelet count < 200 x 10(9)/l mobilized as many CD34+ cells with granulocyte colony-stimulating factor (G-CSF) alone as with chemotherapy and haematopoietic growth factors. Increasing patient age had no effect on post-transplant neutrophil recovery, but significantly delayed platelet recovery (>/= 50 x 10(9)/l) if < 2 x 10(6)/kg CD34+ cells were infused, but this effect was eliminated completely with infusion of >/= 4 x 10(6)/kg CD34+ cells. Increasing age adversely affected CD34+ yield even with limited premobilization therapy, indicating that early collection is important in elderly patients. PMID- 12580956 TI - Follicular lymphoma lacking the t(14;18)(q32;q21): identification of two disease subtypes. AB - The clinical and pathological features, including karyotype data and BCL2 protein expression pattern, of follicular lymphoma without a t(14;18)(q32;q21) have not been well defined. We have identified and conducted a detailed analysis of 50 cases with follicular lymphoma who lack the t(14;18). Fluorescent in situ hybridization (FISH) analysis was used to exclude cases with a cryptic IGH/BCL2 rearrangement. BCL2 protein expression level was assessed by immunohistochemistry. The karyotypes were assessed for recurrent sites of structural rearrangement, duplications and deletions on a band-by-band basis, and compared with a large cohort of cases with t(14;18). A distinct pattern of chromosomal alterations was identified in the cases without t(14;18). BCL2 protein overexpression was detected in 33% of 49 tested cases. In this minority, the karyotypes frequently showed increased copies of chromosome 18. The majority of cases (67%) did not show BCL2 overexpression and were characterized prominently by the presence of t(3;14)(q27;q32), implying a role for BCL6. Follicular lymphomas that lack a t(14;18) were segregated into two subgroups with distinct cytogenetic, phenotypic and possibly clinical features: one with BCL2 protein overexpression not related to an IGH/BCL2 rearrangement and a second without BCL2 overexpression. Objective identification of BCL2 expression level as well as BCL2 and BCL6 status by cytogenetic or FISH analysis has potential clinical utility and may yield insights into alternative genetic mechanisms associated with B-cell lymphomas with a follicular growth pattern. PMID- 12580957 TI - Abnormal expression of apoptosis-related genes in haematological malignancies: overexpression of MYC is poor prognostic sign in mantle cell lymphoma. AB - The expression of apoptosis-related genes BCL2, BAX, BCL2L1, BCL2A1, MCL1, DAPK1 and MYC was studied by quantitative real-time polymerase chain reaction on total RNA samples from patients with acute lymphoblastic leukaemia (ALL, n = 16), acute myeloid leukaemia (AML, n = 27), chronic myeloid leukaemia (CML, n = 12), mantle cell lymphoma (MCL, n = 19) and chronic lymphoid leukaemia (CLL, n = 32). BCL2, BAX, BCL2A1, MCL1, DAPK1 and MYC were overexpressed in all patient groups. BCL2L1 was underexpressed in CLL and CML, but not in AML, ALL and MCL. MCL1 levels were significantly higher in CD13 and CD33-positive ALL, and in CD56-positive AML samples. BCL2, BCL2L1, BCL2A1 and MCL1 were overexpressed and DAPK1 was underexpressed in CLL samples with a 11q23 deletion. MYC overexpression was significantly associated with shorter overall survival in MCL (P < 0.01). AML patients with a normal karyotype showed a higher frequency of BCL2A1 overexpression (P < 0.001) than those with an abnormal karyotype. PMID- 12580958 TI - Expression of co-stimulatory molecules on acute myeloid leukaemia blasts may effect duration of first remission. AB - Many solid tumours have been shown to lack expression of either of the immune co stimulatory molecules CD80 (B7.1) or CD86 (B7.2), which is thought to be one of the ways in which tumours may escape immune recognition. We have examined the surface expression of CD80, CD86, human leucocyte antigen (HLA) class I and II, CD11a, CD54, and CD58 on the blast cells from patients with acute myeloid leukaemia (AML) at presentation. CD80 was only rarely expressed on AML blasts and, in those leukaemic cells expressing CD80, the level of expression was low. In contrast, expression of CD86 was detected on the AML blasts in more than half of the samples tested and, in some cases, the level of expression was equivalent to that of mature monocytes and activated B lymphocytes. The percentage of leukaemic blasts expressing CD86 was higher in the M4 and M5 French-American British (FAB) types, and expression of CD11a and HLA class II was higher in the M4 FAB type. There was no difference in expression of CD80, CD54, CD58, or HLA Class I between different FAB subgroups. There was no significant difference in duration of first remission with expression of CD80, CD86, CD11a, CD54 or HLA class II. However, when expression of CD80 and CD86 were considered together, a significantly longer duration of remission was found. We suggest that these molecules may play a role in immunosurveillance, resulting in prolonged remission in some patients treated for AML. PMID- 12580959 TI - Plasma interleukin 8 level predicts for survival in chronic lymphocytic leukaemia. AB - The malignant B cells of patients with chronic lymphocytic leukaemia (CLL) constitutively express interleukin 8 (IL-8) and IL-8 receptors. Ex vivo culture with exogenous IL-8 enhances IL-8 expression and prolongs leukaemia cell survival, partly through increased bcl-2 expression. IL-8 may function as an autocrine growth and apoptosis resistance factor in CLL. Therefore, we evaluated the prognostic relevance of plasma IL-8 levels in 151 CLL patients [median age 61 years (range, 32-84 years), median plasma IL-8 level 18.9 pg/ml (9.1-89.1 pg/ml)]. All Rai stages were represented; advanced stage was associated with significantly higher plasma IL-8 levels (P < 0.0001, Kruskal-Wallis). Also, plasma IL-8 level was correlated with serum beta2-microglobulin (beta2-M) (R = 0.24, P = 0.0081), haemoglobin (R = -0.39, P < 0.0001) and platelet count (R = 0.23, P = 0.0049) by Spearman's rank correlation. Univariate analysis using Cox proportional hazards models identified elevated IL-8 and beta2-M as significant prognostic factors with relative risks of 7.43 (P = 9.1 x 10(-9)) and 16.40 (P = 5.9 x 10(-10)) respectively. High levels of IL-8 were associated with shorter survival independent of beta2-M level. Using recursive-partitioning procedures, an IL-8 cut-off point of 26.2 pg/ml segregated a group of CLL patients with significantly shorter survival (median 9.3 months) (P < 0.0001). In conclusion, plasma IL-8 level in CLL patients correlates with other prognostic factors, such as Rai stage and beta2-M, and is associated with increased risk of death in CLL patients. The role of IL-8 inhibitors in the treatment of patients with CLL should be explored. PMID- 12580960 TI - Overexpression of manganese superoxide dismutase promotes survival in cell lines after doxorubicin treatment. AB - Overexpression of manganese superoxide dismutase (MnSOD) has been postulated as one possible mechanism of protection from oxidative damage and free radicals. Doxorubicin treatment induces oxygen free radicals, leading to cytotoxicity and myelosuppression. The present study was performed to determine whether over expression of MnSOD may play a role in resistance to doxorubicin. Retroviral constructs having the human MNSOD gene in the sense orientation and the neomycin phosphotransferase gene (NEOR) as a selectable marker were transduced into the human melanoma cell line A375 and the human histiocytic lymphoma cell line U937. Stably transduced A375 and U937 cells were subjected to 10-100 ng/ml doxorubicin for 24 h and compared with doxorubicin-treated A375 and U937 cells transduced with vector only. A colony forming assay was used to determine cell viability in semi-solid medium. Results demonstrated that wild-type A375 and U937 cells display low levels of endogenous MnSOD mRNA and protein, and are sensitive to doxorubicin treatment. In contrast, A375 and U937 cells transduced with the MNSOD gene consistently demonstrate increased colony formation in the presence of increasing concentrations of doxorubicin. MnSOD-transduced A375 and U937 cells also demonstrate increased MnSOD mRNA and protein levels when compared with wild type or those cells transduced with vector only. These results indicate that overexpression of MnSOD can enhance resistance to doxorubicin treatment. PMID- 12580961 TI - Mutational analysis of class III receptor tyrosine kinases (C-KIT, C-FMS, FLT3) in idiopathic myelofibrosis. AB - Genomic DNA from patients with idiopathic myelofibrosis (IMF) was screened by polymerase chain reaction (PCR) and conformation sensitive gel electrophoresis (CSGE) for mutations in the C-KIT gene (60 patients), as well as the C-FMS and FLT3 genes (40 patients). Intronic primers were used to amplify the entire coding region of both the C-KIT and C-FMS genes, and selected regions of the FLT3 gene. CSGE and direct DNA sequencing detected all previously reported as well as several novel polymorphisms in each of the genes. A novel c-fms exon 9 mutation (Gly413Ser) was detected in two patients. Its functional significance remains to be determined. The c-kit mutation Asp52Asn, previously described in two of six IMF patients in Japan, was not detected in this study. In addition, the reported c-fms mutations involving codons 301 and 969 were not identified. Therefore, in contrast to acute myeloid leukaemia, mutations in RTKs class III do not appear to play a significant pathogenetic role in idiopathic myelofibrosis. PMID- 12580962 TI - Detection of human herpesvirus-8 in peripheral blood mononuclear cells from adult Japanese patients with multicentric Castleman's disease. AB - Human herpesvirus-8 (HHV-8) encodes viral homologues of cellular genes, including viral interleukin 6 (vIL-6), which induces endogenous human IL-6 (hIL-6) secretion. Unregulated overproduction of hIL-6 in lymph nodes (LN) is thought to be responsible for the systemic manifestations of multicentric Castleman's disease (MCD). In the present study, we assessed the presence of HHV-8 and HHV-8 encoded viral homologues in LN and peripheral blood mononuclear cells (PBMC) from adult Japanese patients with MCD. HHV-8 DNA was amplified by nested polymerase chain reaction (PCR) and was detected in LN from 13 out of 16 MCD patients (81%). HHV-8 DNA was also detected in PBMC from six out of seven patients (86%) whose LN were positive for HHV-8 DNA. Because mRNA could not be successfully extracted from LN sections that were either formalin-fixed or embedded in paraffin, we examined the expression of mRNA for HHV-8-encoded viral homologues, such as vIL 6, vBCL-2, vCyclin-D and viral G-protein-coupled receptor (vGPCR) by nested reverse transcription (RT)-PCR in PBMC from 10 MCD patients. However, mRNA of these HHV-8-encoded viral homologues was not detected in any patients tested. Although our results do not indicate a role for HHV-8-encoded viral homologues in the pathogenesis of MCD, they do suggest that HHV-8 infection may be associated with MCD in adult Japanese patients. PMID- 12580963 TI - Expansion of CD8+ T cells that express low levels of the B cell-specific molecule CD20 in patients with multiple myeloma. AB - An expanded cytotoxic/memory T-cell subpopulation expressing low levels of the B cell-specific CD20 molecule was found in peripheral blood and bone marrow of patients with multiple myeloma at the time of diagnosis, but returned to normal levels following treatment. CD3+CD20dim cells were also increased in monoclonal gammopathy of unknown significance albeit at lower levels. Lower CD3+CD20dim cell numbers at baseline may be associated with lack of response to treatment and a poor outcome. Because expansion of these T cells may be related to disease status, further studies should investigate their potentially unique function in plasma cell dyscrasias. PMID- 12580964 TI - Absence of SCL mutations in myeloid malignancies. AB - Transcription factors play a central role in the pathogenesis of acute leukaemia. Genes encoding transcription factors are frequently perturbed by chromosome rearrangements. More recently acquired point mutations have been described in a number of important genes for normal haematopoiesis. The stem cell leukaemia (SCL) gene encodes a basic helix-loop-helix transcription factor and is upregulated by chromosome rearrangements in T-acute lymphoblastic leukaemia. SCL function is essential for the development of both primitive and definitive haematopoiesis. We searched for SCL mutations in myeloid leukaemias and chronic myeloproliferative disorders. Our data demonstrated that SCL point mutations were rare in acute myeloid leukaemia and myeloproliferative disorders. PMID- 12580965 TI - E2A-PBX1 fusion in adult acute lymphoblastic leukaemia: biological and clinical features. AB - Molecular and cytogenetic studies performed in 305 adult acute lymphoblastic leukaemia (ALL) patients enrolled in the gimema (Gruppo Italiano Malattie EMatologiche dell'Adulto) multicentric protocols identified an E2A-PBX1 fusion and/or t(1;19) in 10 patients (3.3%). All had common ALL, were mostly CyIg+ and were CD34/CD13/CD33-. Nine patients achieved a complete remission (CR); five patients showed a haematological relapse after 7 months (median). Four patients are alive in first CR with a median follow-up of 29 months; three patients are molecularly negative. This abnormality is frequently associated with early treatment failure. E2A-PBX1+ adult ALL should be considered for intensified treatment strategies and monitoring of minimal residual disease. PMID- 12580966 TI - Common clonal T-cell origin in a patient with T-prolymphocytic leukaemia and associated cutaneous T-cell lymphomas. AB - An unusual course was observed in a patient with indolent T-prolymphocytic leukaemia (T-PLL) who subsequently developed mycosis fungoides (Mf), lymphomatoid papulosis (LyP) and cutaneous CD30+ anaplastic large cell lymphoma (ALCL). Polymerase chain reaction analysis demonstrated identical monoclonal T-cell receptor-beta and -gamma gene rearrangements in all the different clinical entities. Furthermore, cytogenetic studies revealed the same aberrant clone with trisomy of chromosome 8 in T-PLL and ALCL cells. This unique observation suggests that in T-PLL, the leukaemic cells might undergo secondary transformation, subsequently resulting in different phenotypes of cutaneous T-cell lymphoma. PMID- 12580967 TI - Re-entry of tumour B cells into the cycle of somatic mutation and isotype switching in follicular lymphoma. AB - Sequencing of the immunoglobulin (Ig) gene transcripts of the tumour B cells in lymph node (LN) and bone marrow (BM) from a follicular lymphoma (FL) patient associated with multiple myeloma identified two dominant clones. One of the clones, present in both LN and BM, had somatic mutations and extensive clonal diversity. Among the diversified clones, two dominant populations of identical sequences (group I and II) were present. Group II was a descendant population of group I and had nine more somatic mutations. Group I contained micro-, delta-, gamma- and alpha-expressing clones. Group II clones contained mainly micro- and delta-expressing clones. These findings showed that somatic mutation and isotype switching occurred repeatedly in this patient. PMID- 12580968 TI - Cell surface CD43 determination improves diagnostic precision in late B-cell diseases. AB - Panels of immunological markers are useful in refining diagnosis in view of certain variability between B-cell leukaemias. A statistical multivariate approach was used on 100 B leukaemias (preliminary sample) to explore the potential value of the combination of CD43, and the classical markers CD5, CD23, CD79b, FMC7, CD22 and surface immunoglobulin to differentiate chronic lymphoid leukaemia (CLL) from lymphoma (non-CLL). CD43 was highly effective (P < 0.00001) and its inclusion in the panels improved the accuracy of discrimination in a 'control' sample of 74 B leukaemias to 98.6%. Inclusion of CD43 facilitates the diagnosis of B-lymphoproliferative disorders and improves their classification. PMID- 12580969 TI - Imatinib mesylate (STI571) treatment in patients with chronic-phase chronic myelogenous leukaemia previously submitted to autologous stem cell transplantation. AB - Imatinib mesylate (STI571) is a highly effective and well-tolerated treatment for patients with chronic-phase chronic myeloid leukaemia (CML), but information on its efficacy and tolerance in intensively pretreated patients is scarce. Thirty three chronic-phase CML patients who were resistant or intolerant to interferon (IFN) and had been previously submitted to autologous stem cell transplantation were treated with imatinib for a median of 14 months (range: 6-19 months). Seven patients were in haematological response (HR) at the start of treatment; the remaining 26 attained a HR at a median of 3 weeks (range: 1-4 weeks). Major cytogenetic response rates at 3, 6 and 12 months were 42%, 45% and 55%, respectively, including 21%, 24% and 33% complete responses. Grade 3-4 neutropenia, thrombocytopenia and anaemia developed in 33%, 27% and 12% of patients respectively. Non-haematological toxicity included superficial oedema (21% of patients), gastrointestinal symptoms (18%), muscle cramps (15%), skin rash and liver enzyme increase (3% each). These results were not significantly different from those in 65 chronic-phase CML patients, resistant or intolerant to interferon without a previous ASCT, who were included in the same protocol. Imatinib mesylate is effective and safe in chronic-phase CML patients with a previous history of intensive treatment. PMID- 12580970 TI - In vivo imaging studies of the effect of recipient conditioning, donor cell phenotype and antigen disparity on homing of haematopoietic cells to the bone marrow. AB - Homing of transplanted bone marrow cells (BMC) to the host bone marrow (BM) is the first step of engraftment towards durable multilineage haematopoietic reconstitution. We used an in vivo assay to track PKH-labelled cells in the BM of mice after transplantation, using fluorescence microscopy through an optical window placed over the distal femoral epiphysis. Within hours after intravenous injection, the cells moved in and out the femur, and were mobile within the marrow space. One hour after injection of whole BMC into non-conditioned syngeneic and allogeneic recipients, the homing efficiencies (HE) were 1.23 +/- 0.14% and 0.12 +/- 0.02% respectively (P < 0.001). Irrespective of antigen disparity, the number of PKH-labelled cells in the femur decreased by 30% and 50% after 1 and 3 d respectively (P < 0.001). Similar HE of naive and irradiated cells suggested that the majority of cells (> 80%) were quiescent in the BM during the first 3 d. HE were twofold higher in busulphan-myeloablated recipients (P < 0.001 vs non-conditioned), and allogeneic transplantation resulted in 84 +/- 9% donor chimaerism at 4 weeks. The HE of lin- cells was 16-fold higher than that of lin+ cells (P < 0.001), and the subset of lin- SCA-1+ cells was 4.6-fold higher in the BM-homed cell population (P < 0.001 vs lin- cells). Approximately 1,500 of the BM-homed cells rescued 62-71% secondary syngeneic and allogeneic myeloablated recipients. Strikingly, the HE could be predicted during the first 3 d after transplantation by correcting the measurements performed in vivo for the enrichment of progenitors in donor inoculum, donor-recipient antigen disparity and myeloablative conditioning. PMID- 12580971 TI - The Japanese cord blood bank network experience with cord blood transplantation from unrelated donors for haematological malignancies: an evaluation of graft versus-host disease prophylaxis. AB - Cryopreserved umbilical cord blood (CB) from unrelated donors can restore haematopoiesis after myeloablative therapy in patients with haematological malignancy. We investigated the clinical outcomes of CB transplantation (CBT) with special emphasis on graft-versus-host disease (GVHD) prophylaxis. Patients with haematological malignancies (n = 216) received intensive chemotherapy or immunosuppressive therapy, followed by transplantation of cryopreserved CB cells from unrelated donors. The clinical outcomes, i.e. haematological reconstitution, the incidence of acute or chronic GVHD, relapse and event-free survival (EFS), were evaluated. The estimated probability of neutrophil recovery was 88.2%. The median follow-up for the survivors was 557 d (range 21-1492 d). The overall and EFS rates were 32.6% and 25.5%, respectively, 3.5 years after transplantation. Multivariate analysis using Cox's proportional hazards model showed that high risk disease status at CBT and single-drug GVHD prophylaxis were associated with worse 2-year EFS rates [P = 0.0013, relative risk (RR) 1.90, 95% confidence interval (CI) 1.28-2.81 and P = 0.0007, RR 1.91, 95% CI 1.31-2.79 respectively). Age at CBT had no significant influence on EFS. Cryopreserved CB from unrelated donors can restore haematopoiesis in patients with haematological malignancy. Although the incidence is low, the prophylaxis for acute GVHD is an important factor for survival of CBT from unrelated donors. A high rate of suitable donors was found, with a probability of 1 to every 18 CB units, when compared with human leucocyte antigen matching at other haematopoietic stem cell banks. PMID- 12580972 TI - Complete remission of tumour with interleukin 2 therapy in a patient with non Hodgkin's lymphoma post allogeneic bone marrow transplant associated with polyclonal T-cell bone marrow lymphocytosis. AB - A patient with low-grade non-Hodgkin's lymphoma (NHL) who relapsed shortly after an allogeneic bone marrow transplant (BMT) is reported. The patient was treated with interleukin 2 (IL-2), which resulted in a flare-up of graft-versus-host disease followed by disease control, with disappearance of peripheral lymphadenopathy. Sequential bone marrow testing showed the disappearance of bone marrow involvement with disease but occurrence of T-cell aggregates post IL-2 that were identified as polyclonal by molecular methods. The patient remains in complete remission 37 months following allogeneic BMT. PMID- 12580973 TI - Oral anticoagulants in the elderly. AB - Over a 1-year period, 34,998 prothrombin time tests were performed on 2,379 patients aged 40-89 years with a recommended target International Normalized Ratio (INR) value of 2.5. At least one INR value of >/= 5 was found in 507/2,379 patients (21.3%). The elderly demonstrated higher maximum and lower minimum INR values, lower warfarin doses, and an increased number of tests performed. After adjustment for these factors, the risk of INR values >/= 5 increased with age by 15% every 10 years (95% CI 4-28%). We conclude that age is a risk factor for more unstable prothrombin time results. PMID- 12580974 TI - Paradoxical association between the 316 Trp to Ser beta 2-glycoprotein I (Beta2GPI) polymorphism and anti-Beta2GPI antibodies. AB - We report a woman with an obstetric history of a stillbirth at 28 weeks, associated with hypertension and severe intrauterine growth restriction and a miscarriage at 9 weeks. She was persistently positive for immunolgobulin G (IgG) anticardiolipin antibodies and IgG anti-Beta-2-glycoprotein I (anti-Beta2GPI) antibodies. She has delivered three healthy babies when managed antenatally with aspirin and low-molecular-weight heparin prophylaxis. Genotyping revealed that she was homozygous for the 316 Trp to Ser Beta2GPI polymorphism. Studies examining the binding of her plasma Beta2GPI to purified cardiolipin showed markedly reduced binding in comparison with Beta2GPI in pooled normal plasma. PMID- 12580975 TI - L-arginine levels are diminished in adult acute vaso-occlusive sickle cell crisis in the emergency department. AB - Paediatric studies have demonstrated that l-arginine (l-arg), the precursor to nitric oxide, is diminished in vaso-occlusive crisis (VOC). This study aimed to determine whether l-arginine levels are altered in adult VOC in the emergency department. Plasma l-arg and nitric oxide metabolite (NOx) levels were obtained in adult VOC patients presenting to the emergency department. Fifty patients had significantly low plasma l-arg (29.78 micromol/l +/- 11.21, P < 0.05 vs steady state control = 41.16 micromol/l +/- 5.04) and significantly low plasma NOx (12.33 micromol/l +/- 10.28, P < 0.05 vs steady-state control = 25.2 +/- 2.6 micro mol/l). Neither l-arg nor NOx levels could predict VOC clinical course. PMID- 12580976 TI - Hemolink, an o-raffinose cross-linked haemoglobin-based oxygen carrier, does not affect activation and function of human platelets in whole blood in vitro. AB - Haemoglobin-based oxygen carriers (HBOCs) are anticipated to be safe and efficient alternatives to RBC transfusions. Haemoglobin (Hb) raffimer (Hemolink; Hemosol, Toronto, ON, Canada) is polymerized human Hb, cross-linked with o raffinose. As administration of cell-free Hb may affect blood cells and tissues, this study was focused on evaluating effects of Hb raffimer on human platelets in whole blood in vitro. Citrated blood from healthy donors was incubated with Hb raffimer to achieve raffimer concentrations of 2-50 vol percentage (2-50 g/l). Platelet activation, phosphatidylserine exposure and microparticle generation were measured by flow cytometry. Aperture closure time on collagen/ADP- and collagen/epinephrine-coated membranes was determined by a platelet function analyser (PFA-100). We found that addition of Hb raffimer to blood samples up to 50 vol % did not affect human platelets as measured by various markers of platelet activation (CD42b, CD41, PAC-1, CD62, CD63), procoagulant activity (annexin V) and microparticle formation; differences between Hb raffimer- and lactated Ringer's-diluted blood were not significant. Similarly, no adverse effect of Hb raffimer on closure time was observed at concentrations up to 50 vol %, in comparison with Ringer's solution. These data indicate that exposure of human blood to high concentrations of Hb raffimer in vitro did not cause platelet activation nor affect platelet function. PMID- 12580977 TI - GATA-1 transcription factor is mutated in CMK megakaryoblastic cell line. PMID- 12580978 TI - The optimal management of polycythaemia vera. PMID- 12580982 TI - Pharmacokinetics, food intake requirements and tolerability of once-daily combinations of nelfinavir and low-dose ritonavir in healthy volunteers. AB - AIMS: This study was performed to evaluate the steady-state pharmacokinetics, food intake requirements and short-term tolerability of once-daily combinations of nelfinavir and low-dose ritonavir. METHODS: Twenty-seven healthy volunteers were randomized over three groups to receive a once-daily regimen of nelfinavir/ritonavir 2,000/200 mg (group 1), 2,000/400 mg (group 2) or 2,500/200 mg (group 3) with food for 14 days. Pharmacokinetic parameters for nelfinavir and its active metabolite M8 were assessed on study days 15 and 16, after administration of the regimens with a full (610 kcal) or light (271 kcal) breakfast, respectively. RESULTS: Pharmacokinetic data were evaluable for eight volunteers in group 1, eight in group 2 and four in group 3. Administration of nelfinavir/ritonavir with a full breakfast resulted in geometric mean (GM) nelfinavir AUC(24h) values of 76.8, 51.3, and 61.9 h*mg/l in group 1, 2 and 3, respectively. GM 24-h Cmin concentrations of nelfinavir were 0.76 mg l(-1), 0.43 mg l(-1) and 0.47 mg l(-1), respectively. Co-administration of ritonavir increased M8 concentrations more than nelfinavir concentrations, resulting in GM AUC(24h) and Cmin values for nelfinavir plus M8 that were higher than or comparable to reference values for the approved regimen of nelfinavir (1,250 mg BID without ritonavir). In the 2,000/200 mg group, seven out of eight subjects had a Cmin value of nelfinavir plus M8 above a threshold of 1.0 mg l-1. Administration of the combinations with a light breakfast resulted in significant decreases in the AUC(24h) and Cmin of nelfinavir and nelfinavir plus M8, compared with intake with a full breakfast. For the Cmin of nelfinavir plus M8, the GM ratio (light/full breakfast) was 0.76 (90% confidence interval 0.67-0.86, participants from all groups combined). Short-term tolerability was satisfactory, apart from a higher than expected incidence of mild rash (12%). CONCLUSIONS: Administration of nelfinavir in a once-daily regimen appears feasible. A nelfinavir/ritonavir 2,000/200 mg combination appears appropriate for further evaluation. Once-daily nelfinavir/ritonavir should be taken with a meal containing at least 600 kcal. PMID- 12580983 TI - Comparative study of analgesic efficacy and morphine-sparing effect of intramuscular dexketoprofen trometamol with ketoprofen or placebo after major orthopaedic surgery. AB - AIMS: Multimodal analgesia is thought to produce balanced and effective postoperative pain control. A combined therapy with nonsteroidal anti inflammatory drugs (NSAIDs) and opiates could result in synergistic analgesia by acting through different mechanisms. Currently there are very few parenterally administered NSAIDs suitable for the immediate postoperative period. Therefore, this study was undertaken to assess the analgesic efficacy, relative potency, and safety of parenteral dexketoprofen trometamol following major orthopaedic surgery. METHODS: One hundred and seventy-two patients elected for prosthetic surgery, were randomized to receive two intramuscular injections (12 hourly) of either dexketoprofen 50 mg, ketoprofen 100 mg or placebo in a double-blind fashion. Postoperatively, the patient's pain was stabilized, then they were connected to a patient- controlled analgesia system (PCA) of morphine for 24 h (1 mg with 5 min lockout). RESULTS: The mean cumulative amount of morphine (CAM) used was of 39 mg in the dexketoprofen group and 45 mg in the ketoprofen group vs 64 mg in the placebo group. (Reduction in morphine use was approximately one third between the active compounds compared with placebo (adjusted mean difference of -25 mg between dexketoprofen and placebo and -23 mg between ketoprofen and placebo. These differences were statistically significant: P /= 65 years) modified the risk of having minor GI events. During treatment or within 1 month of stopping, 110 serious GI events were reported (including 76 upper GI bleeds/peptic ulcers, one perforated colon), 101 thromboembolic events, three reports of acute renal failure, one each of Stevens-Johnson syndrome, severe anaphylaxis and angio-oedema. CONCLUSIONS: Doctors should continue to prescribe NSAIDs including COX-2 selective inhibitors with caution. PMID- 12580990 TI - The effect of ketoconazole on the in vivo intestinal permeability of fexofenadine using a regional perfusion technique. AB - AIMS: To investigate whether the drug-drug interaction between fexofenadine and ketoconazole is localized to efflux transport proteins of the small intestine, and to determine and classify the effective jejunal permeability (Peff) of fexofenadine according to the Biopharmaceutics Classification System (BCS). METHODS: Two separate jejunal perfusion experiments were performed using the Loc I-Gut technique in eight healthy volunteers. During treatment 1 (T1), we investigated the acute effect of ketoconazole on the Peff and plasma pharmacokinetics of fexofenadine. In treatment 2 (T2) we examined the effect of oral pretreatment with ketoconazole (200 mg daily for 5 days) on the same absorption parameters. Each experiment was divided into two periods of 100 min and the jejunal segment was perfused with 93 micro m fexofenadine during both periods. In period 2 of each treatment, fexofenadine was coadministered with 94 micro m ketoconazole. The concentrations of fexofenadine in intestinal perfusate and plasma were measured by liquid chromatography with mass detection. RESULTS: During T1, the mean (+/- s.d.) Peff of fexofenadine was low according to the BCS (0.11 +/- 0.11 and 0.04 +/- 0.13 x 10(-4) cm s(-1) in periods 1 and 2, respectively), and the coadministration of ketoconazole in period 2 had no significant acute effect on Peff (95% confidence interval (CI) on the difference 0.37, 0.51). After pretreatment with ketoconazole (T2), the jejunal Peff of fexofenadine increased to 0.29 +/- 0.47 and 0.22 +/- 0.31 x 10-4 cm s(-1) in both periods 1 and 2, respectively, but the change was not statistically significant when compared with T1 (95% CI on the difference -0.62, 0.27 for T1 0-100 min vs T2 0-100 min; -0.54, 0.34 for T1 0-100 min vs T2 100-200 min). Fexofenadine plasma AUC from 0-100 mg showed no significant difference after pretreatment with ketoconazole (55 +/- 101 and 51 +/- 33 micro g ml(-1) min(-1) respectively; 95% CI on the difference -108, 115). Total plasma AUC (0-720 min) was 318 +/- 426 and 426 +/- 232 ng ml(-1) min in T1 and T2, respectively (95% CI on the difference 622, 405). CONCLUSIONS: No significant effect of acute coadministration or pretreatment with ketoconazole on the in vivo intestinal absorption of fexofenadine was detected in this study. PMID- 12580991 TI - Interaction of ibuprofen and probenecid with drug metabolizing enzyme phenotyping procedures using caffeine as the probe drug. AB - AIMS: To examine the suspected inhibitory potential of the over-the-counter (OTC) drug ibuprofen on N-acetyltransferase 2 (NAT2) in vitro and in vivo and the possible implications for phenotyping procedures using caffeine as probe drug. METHODS: We first studied the inhibitory effect of ibuprofen on NAT2 in vitro, using human liver cytosol and sulfamethazine as substrate. In vivo 15 fast and 15 slow acetylating healthy volunteers were treated with a single dose of ibuprofen (800 mg) orally and phenotyped for NAT2, CYP1A2, and xanthine oxidase (XO) with caffeine as probe drug before and during drug treatment. Because of unexpected in vivo results with ibuprofen this study was repeated in 20 healthy volunteers with probenecid, a model substrate of renal organic anion transport (OAT). For phenotyping tests a urine sample was collected 6 h after caffeine (200 mg) intake. The caffeine metabolites acetyl-6-formylamino-3-methyluracil (AFMU), 1 methylxanthine (1MX), 1-methyluric acid (1MU), and 1,7-dimethyluric acid (17MU) were quantified by HPLC, and the corresponding metabolic ratios for CYP1A2, NAT2, and XO were then calculated. Genotyping for NAT2 was performed with standard PCR RFLP methods. RESULTS: In vitro, with human liver cytosol an inhibition by ibuprofen of the acetylation of sulfamethazine with Ki values between 2.2 and 3.1 mm was observed. Surprisingly, in vivo a significant (P < 0.001) increase of the acetyl-6-formylamino-3-methyluracil/1-methylxanthine (AFMU/1MX) urinary ratio from 0.97 +/- 0.16 to 1.08 +/- 0.18 (95% CI on the difference 0.049, 0.170) was found, indicating an apparent elevation of NAT2 activity. In contrast, no change was observed for the ratios used for XO and CYP1A2. Because an induction of NAT2 could be excluded, an interaction of ibuprofen with the tubular secretion of some of the caffeine metabolites was assumed. To prove this assumption, the in vivo study was repeated with probenecid, a model substrate of the renal OAT system. Again, a prominent elevation of the AFMU/1MX ratio from 0.97 +/- 0.21 to 1.53 +/- 0.35 was found (P < 0.002; 95% CI on the difference 0.237, 0.876), but also the XO ratio 1MU/1MX was significantly (P < 0.0001) increased from 1.34 +/- 0.09 to 2.24 +/- 0.14 (95% CI on difference 0.735, 1.059) due to a reduction of 1MX excretion. CONCLUSIONS: Substrates of OAT interact with renal excretion of caffeine metabolites and may falsify NAT2 and XO phenotyping results. Other phenotyping procedures, which are based on urinary metabolic ratios, should also be validated in this respect, especially in patients under polymedication. PMID- 12580992 TI - Effect of short-term administration of garlic supplements on single-dose ritonavir pharmacokinetics in healthy volunteers. AB - AIMS: To evaluate the effect of acute dosing of garlic supplements on the single dose pharmacokinetics of ritonavir. METHODS: Ten healthy volunteers (five male, five female) were equally randomized in a crossover design to receive 400 mg of a single dose of ritonavir within 10 min after eating breakfast either alone or with 10 mg of Natural Source Odourless Garlic. They received a total of eight doses of garlic extract (2 x 5 mg capsules) taken twice daily for 4 days. Ritonavir and the seventh garlic dose were administered simultaneously. RESULTS: Coadministration of garlic nonsignificantly decreased area under the plasma concentration-time curve (AUC(0, infinity )) by -17% (90% confidence interval (CI), -31% to 0%; range -46% to 68%) and peak plasma concentration of ritonavir by -1% (90% CI, -25% to 31%; range -51% to 136%). CONCLUSIONS: Acute dosing of the garlic capsules over 4 days did not significantly alter the single-dose pharmacokinetics of ritonavir in healthy volunteers. Given the complex effects of both ritonavir and garlic on drug metabolism, the results of our study should not be extrapolated to steady-state conditions, where the possibility of an interaction still needs to be evaluated. PMID- 12580993 TI - Alterations in cyclosporin A pharmacokinetics and metabolism during treatment with St John's wort in renal transplant patients. AB - AIM: This study investigated the effects of St John's wort extract (SJW) on the pharmacokinetics and metabolism of the immunosuppressant cyclosporin A (CSA). METHODS: In an open-label study, 11 renal transplant patients received 600 mg SJW extract daily for 14 days in addition to their regular regimen of CSA. Blood concentrations of CSA and its metabolites AM1, AM1C, AM9, AM19, and AM4N were measured by HPLC. RESULTS: After 2 weeks of SJW coadministration, dose-corrected AUC0-12, Cmax and Ctrough values for CSA decreased significantly by 46%[geometric mean ratio baseline/SJW (95% CI): 1.83 (1.63-2.05)], 42%[1.72 (1.42-2.08)], and 41%[1.70 (1.17-2.47)], respectively. CSA doses were increased from a median of 2.7 mg day(-1) kg(-1) at baseline to 4.2 mg day(-1) kg(-1) at day 15, with the first dose adjustment required only 3 days after initiation of SJW treatment. Additionally, the metabolite pattern of CSA was substantially altered during SJW treatment. Whereas dose-corrected AUC values for AM1, AM1c and AM4N significantly decreased by 59%, 61%, and 23% compared with baseline, AUC values for AM9 and AM19 were unchanged. Following the increase in CSA dose, observed AUC and Cmax values for AM9, AM19, and AM4N increased by 20-51% and 43-90%, respectively. CONCLUSION: Administration of SJW extract to patients receiving CSA treatment resulted in a rapid and significant reduction of plasma CSA concentrations. Additionally, the substantial alterations in CSA metabolite kinetics observed may affect the toxicity profile of the drug. PMID- 12580995 TI - A population-based study of maternal use of amoxicillin and pregnancy outcome in Denmark. AB - AIMS: Amoxicillin is a widely used penicillin, but data on its safety in pregnancy are limited. We examined the association between amoxicillin exposure during pregnancy and birth weight, preterm delivery, congenital malformations, perinatal death, and spontaneous abortion. METHODS: We identified all primiparous women with a live birth, or a stillbirth after the 28th gestational week, from 1 January 1991-31 December 2000 in the County of North Jutland, Denmark. Data on prescriptions for amoxicillin and outcome were obtained from population-based registries. Using a follow-up and a case-control design, we compared pregnancy outcomes between women who had been prescribed amoxicillin during pregnancy and those who had not, adjusting for available potentially confounding factors. RESULTS: We identified 401 primiparous women who redeemed a prescription for amoxicillin during their pregnancy. The control group consisted of 10 237 primiparous women who did not redeem any prescriptions from 3 months before pregnancy until the end of pregnancy. The adjusted mean birth weight of children born to amoxicillin-exposed mothers was 57 g [95% confidence interval (CI) 9, 105] higher than that of children born to controls. Odds ratios among amoxicillin exposed relative to controls were: low birth weight 0.63 (95% CI 0.26, 1.53), preterm delivery 0.77 (95% CI 0.49, 1.21), congenital malformation 1.16 (95% CI 0.54, 2.50), and spontaneous abortion 0.89 (95% CI 0.66, 1.18). We did not observe any cases of perinatal death in the amoxicillin-exposed women. CONCLUSIONS: We did not find any increased risk of adverse pregnancy outcome associated with amoxicillin exposure during pregnancy, but additional studies are warranted. PMID- 12580994 TI - Low-dose atorvastatin therapy does not augment endothelial function in active hypercholesterolaemic males. AB - AIMS: As statin therapy has been demonstrated to augment endothelial function in sedentary hypercholesterolaemia (HC), we aimed to investigate the effects of atorvastatin therapy on endothelial function in physically active, HC men. METHOD AND RESULTS: Eleven physically active, HC males were recruited. Endothelial function [forearm blood flow response to brachial artery infusion of acetylcholine (Ach)] was assessed twice in each subject following atorvastatin or no therapy in a randomized crossover design. In addition, endothelial function was compared with an active, normolipidaemic control group (C). Atorvastatin therapy reduced total and LDL cholesterol, but had no effect on basal blood flow or endothelial function (peak ACh mean difference +/- standard error 0.75 +/- 1.75 ml min(-1) per 100 ml tissue) [95% confidence interval (CI) -3.1, 4.6]. In addition, there was no difference in endothelial function between the HC and C groups (-1.14 +/- 2.60 ml min(-1) per 100 ml tissue; CI -6.53, 4.25). CONCLUSION: Statin therapy in HC patients with normal endothelial function does not augment endothelial function. PMID- 12580996 TI - Robotic radical retropubic prostatectomy. PMID- 12580997 TI - Penile prostheses: are they still relevant? PMID- 12580998 TI - Opportunities and outcomes in early prostate cancer: a commentary on policy, patient perspectives, trials and strategies. PMID- 12580999 TI - How is shared-care growing up? PMID- 12581000 TI - Incidence of physician-diagnosed interstitial cystitis in Olmsted County: a community-based study. AB - OBJECTIVE: To obtain community-based information about the incidence of interstitial cystitis, a chronic disabling condition of the bladder where knowledge is limited because there are no definitive diagnostic criteria. PATIENTS AND METHODS: All residents of Olmsted County, MN, USA who had received a physician-assigned diagnosis of interstitial cystitis between 1976 and 1996 were identified through the resources of the Rochester Epidemiology Project. The clinical findings at diagnosis and during the follow-up were ascertained from the community medical records for each study subject. RESULTS: In all, 16 women and four men received a diagnosis of interstitial cystitis during the study period. The overall age- and sex-adjusted (95% confidence interval) incidence rate was 1.1 (0.6-1.5) per 100 000 population. The age-adjusted incidence rates were 1.6 per 100 000 in women and 0.6 per 100 000 in men (P = 0.04). The median (range) age at initial diagnosis was 44.5 (27-76) years in women and 71.5 (23-79) years in men (P = 0.26). The median number of episodes of care-seeking for symptoms before the diagnosis was one for women and 4.5 for men (P = 0.03). The median duration from the onset of symptoms until the first diagnosis was 0.06 and 2.2 years in women and men, respectively (P = 0.2). CONCLUSIONS: These findings suggest that the incidence of interstitial cystitis in the community is extremely low. Although the gender difference may be real, the trend toward a later diagnosis in men than in women suggests a potential for missed diagnosis in men. This might explain some of the gender difference in the incidence of interstitial cystitis in men and women. PMID- 12581001 TI - The efficacy of an abbreviated model of the International Prostate Symptom Score in evaluating benign prostatic hyperplasia. AB - OBJECTIVES: To investigate the acceptability to patients in Turkey of the International Prostate Symptom Score (IPSS) for evaluating benign prostatic hyperplasia (BPH), and to devise a possible abbreviated model, saving time when administered by a physician. PATIENTS AND METHODS: The IPSS questionnaire was initially self-administered in 200 consecutive patients aged> 50 years and with lower urinary tract symptoms. Patients were instructed only to mark questions which they clearly understood. The IPSS was then administered to 500 consecutive patients by the same physician. Subsequently, each of the seven questions was separately correlated with the IPSS and quality-of-life (QoL) score. The first five questions with the highest correlation coefficient for both the total IPSS and QoL score were identified, and every possible combination of these questions produced and correlated. RESULTS: In the first group, 29% of the patients did not complete the form and 44% could not mark at least one item; the effect of educational level was significant (P < 0.05). In the second group a combination of the first three items had a Pearson's correlation of 0.90 with the total IPSS; similarly, four questions combined (2, 3, 6 and 7) also correlated well (r = 0.92). Both combinations took significantly less time to administer (P < 0.05). CONCLUSION: The abbreviated form of the IPSS consisting of three or four items may be used to assess symptoms in patients with BPH, especially in underdeveloped countries. PMID- 12581002 TI - Productivity, vitality and utility in a group of healthy professionally active individuals with nocturia. AB - OBJECTIVES: To assess the effect of nocturia on productivity, vitality and utility in a selected group of professionally active individuals with nocturia, compared with matched controls, and investigate the effect of symptom severity, to test the hypothesis that lack of sleep caused by frequent sleep interruptions could reduce an individuals' daytime energy and activity levels. SUBJECTS AND METHODS: Subjects (203) were recruited in Sweden through advertisements, and their suitability for the study assessed in a structured interview. Controls (80) matched for age and gender were randomly selected from a market research panel and given the same interview. Both groups completed a productivity questionnaire, a generic quality-of-life questionnaire with a specific domain for vitality and a utility instrument. RESULTS: The study group with nocturia had a significantly lower level of vitality and utility, and greater impairment of work and activity, than the control group. Women were more affected than men. Symptom severity correlated with all three measures. CONCLUSIONS: In an otherwise healthy and professionally active group of individuals, waking at night to void significantly diminishes their overall well-being, vitality and productivity, leading to a significant level of indirect and intangible costs. PMID- 12581003 TI - Urinary symptoms, quality of life and sexual function in patients with benign prostatic hypertrophy before and after prostatectomy: a prospective study. AB - OBJECTIVE: To evaluate urinary symptoms, sexual dysfunction and quality of life in patients with benign prostatic hypertrophy (BPH) before and after open prostatectomy, using the International Prostate Symptom Score (IPSS), the International Continence Society (ICS)-"BPH" (ICS-male, ICS-sex and ICS-QoL) and International Index of Erectile Function (IIEF) questionnaires. PATIENTS AND METHODS: Sixty men with BPH (mean age 68 years) underwent a digital rectal examination, transurethral ultrasonography, measurement of total prostatic specific antigen serum level and uroflowmetry. Their urinary symptoms, sexual function and quality of life were fully evaluated using the IPSS, ICS-"BPH" and IIEF before and 6 months after suprapubic prostatectomy. The body mass index (BMI) was also calculated for each patient. Univariate analysis was used to examine the relationship between symptom scores and age, tobacco use, alcohol intake and BMI. RESULTS: In a univariate analysis, age was the most important prognostic factor for both urinary and sexual symptoms. Prostatectomy resulted in a significant improvement in obstructive (mean 9.68 to 3.38) and irritative symptom (6.70 to 3.06), and quality-of-life scores (3.41 to 1.34). ICS-male scores were both significantly decreased, the mean voiding score from 13.72 to 10.28 and the incontinence score from 10.43 to 7.81. There was also a significant decrease in the ICS-QoL symptom score (from 9.20 to 7.27). Comparative results between IIEF and ICS-sex showed no improvement in sexual scores after open surgery, but there was a significant increase in sexual desire and overall satisfaction (both P = 0.035). CONCLUSIONS: The combined use of the IPSS, ICS "BPH" and IIEF allows an evaluation of the relationship between age, prostatic symptoms and sexual dysfunction. Age may be considered both a direct and indirect (BPH-related) prognostic factor for sexual activity. Suprapubic prostatectomy resolves obstructive symptoms, and maintains sexual desire, overall sexual satisfaction and an improvement in quality of life. However, irritative symptoms, erection, orgasm and sexual intercourse satisfaction are not significantly altered. PMID- 12581004 TI - Quality control in urodynamics: a review of urodynamic traces from one centre. AB - OBJECTIVE: To investigate quality control in our unit and to enable other units to compare their results, as experience from central reviews of urodynamic traces for multicentre trials has suggested that poor quality control is common. PATIENTS AND METHODS: All consecutive male urodynamic tests conducted over 1 year were reviewed. A list of criteria to assess the quality of the records was devised, based upon International Continence Society guidelines on "good urodynamic practice", and on other sources. Eligible traces were analysed for aspects of quality control, e.g. baseline pressures and coughs to test pressure transmission. The data were analysed to establish how often quality criteria were met, and identify areas for improvement. RESULTS: In 100 eligible traces, the baseline detrusor pressure was 0-10 cmH2O in 86, and - 5 to +10 cmH2O in 94%. Baseline intravesical and abdominal pressure were 30-50 cmH2O in 68% and 73% of cases, respectively. Coughs were present before filling in 94%, during filling in 95%, before voiding in 72% and after voiding in 87% of cases. The cough-test frequency was sufficient in 30% of traces. In 11 the intravesical pressure line fell out during voiding. CONCLUSION: Most of the traces assessed met the quality criteria defined, but significant defects were not uncommon. Some of the problems identified suggest areas of urodynamic technique which should be studied in more detail. We intend to modify our quality control practices, and hope to show an improvement on re-audit. We hope that other urodynamic departments will be encouraged to review their practice, and we aim to improve our results. PMID- 12581005 TI - Knowledge of female bladder care among medical staff, nurses and midwives: results of a questionnaire survey. AB - OBJECTIVE: To determine knowledge about bladder care among junior medical staff and allied health professionals, and to examine any difference in knowledge among the subgroups. SUBJECTS AND METHODS: Using a single-questionnaire survey in a large obstetrics and gynaecology Hospital Trust in an inner city setting, midwives, nurses and medical staff (not consultants) were asked eight questions about different aspects of female bladder physiology and care. The differences in responses among the professional groups were assessed. RESULTS: In all, 120 completed questionnaires were returned (32 doctors, 40 nurses, and 48 midwives). Knowledge was similar on urethral length, bladder capacity, daily fluid intake, ideal size of catheter, catheter balloon size, and the maximum time of use of a short-term catheter. Correct responses for normal daily fluid intake, duration of both short and long-term catheterization were given by fewer than half the respondents. There were significant differences among the professional groups in the correct response rate for long-term catheter duration (P = 0.031) and the normal time interval between voids (P = 0.038). CONCLUSION: There were significant differences in the knowledge of all subgroups about bladder care, and poor levels of knowledge in several areas. This is a potentially serious problem for women at risk of bladder complications after gynaecological surgery and childbirth. All staff involved in managing women after childbirth and gynaecological surgery should have formal training in bladder care, to optimize patient care. PMID- 12581006 TI - Transurethral electrovaporization of the prostate: is it any better than conventional transurethral resection of the prostate? AB - OBJECTIVE: To evaluate transurethral electrovaporization of the prostate (TUVP), compared with transurethral resection of the prostate (TURP), as a treatment for men with symptomatic benign prostatic enlargement (BPE). PATIENTS AND METHODS: In all, 235 men with symptomatic BPE in four hospitals in the South-East of England were randomized to TUVP (115) and TURP (120). Patients were assessed using the International Prostate Symptom Score (IPSS), the Short Form-36 (SF-36), EuroQol and sexual function questionnaire, uroflowmetry, ultrasonographic measurement of residual urine volume, pressure-flow urodynamics and transrectal ultrasonography. RESULTS: There was no statistically significant difference in the objective and subjective outcome after TURP and TUVP. The latter was associated with a lower transfusion rate than TURP but this did not result in an overall reduction in complications. There was no difference in the length of hospital stay. Overall, the two operations produced equivalent results and equivalent complication rates. CONCLUSION: TUVP is an effective treatment for symptomatic BPE, with results equivalent to TURP. TUVP has not led to the expected reduction in early postoperative morbidity or shorter hospital stays. PMID- 12581007 TI - Substitution urethroplasty for anterior urethral strictures: a critical appraisal of various techniques. AB - OBJECTIVE: To retrospectively compare the outcome of various techniques of substitution urethroplasty. PATIENTS AND METHODS: Between 1989 and 2000, 109 patients (mean age 39.5 years) underwent substitution urethroplasty for recurrent anterior urethral strictures. Between 1989 and 1995 the procedure was by ventral placement of free grafts (bladder mucosa, buccal mucosa, penile skin) or penile skin flaps. From 1995 onwards the flaps and grafts (buccal mucosa) were applied either ventrally or dorsally. Stricture recurrence and the complications associated with each technique were compared. RESULTS: Ventral onlay repairs were associated with a higher incidence of complications than dorsal repairs, e.g. postvoid dribbling (39% vs 23%, P = 0.01), ejaculatory dysfunction (20% vs 5%, P = 0.03) and flap/graft pseudo-diverticulum or out-pouching (26% vs 2.6%, P = 0.01). Superficial penile skin necrosis was significantly more common with the use of penile skin flaps than with free grafts. There was no significant difference in stricture recurrence, erectile dysfunction and residual penile deformity among the various techniques. CONCLUSIONS: Dorsal free graft/flap onlay urethroplasty gives better results than ventrally placed free grafts/flaps. Dorsal onlay buccal mucosal urethroplasty is a versatile procedure and associated with fewer complications than other substitution methods. PMID- 12581008 TI - Site-specific positive margins at radical prostatectomy: assessing cancer-control benefits of wide excision of the neurovascular bundle on a side with cancer on biopsy. AB - OBJECTIVE: To determine the potential risk of biopsy-selected nerve-sparing surgery based on the findings of site-specific extracapsular extension (ECE) and positive surgical margins (PSMs) in the area of the neurovascular bundle in radical prostatectomy specimens. PATIENTS AND METHODS: Controlling for surgical technique and pathological interpretation, 221 consecutive patients had their neurovascular bundles removed on the side with a positive biopsy. The surgical specimens were reviewed for ECE and PSM status, specifically in the area of the neurovascular bundle, from apex to base. RESULTS: Of the 221 patients, 38% had ECE and 43 (20%) had a PSM in the area of the neurovascular bundle. This equates to a ratio of 51% for PSM/ECE. An additional 42 men (18%) had ECE with negative margins, but would have been at potential risk for PSMs if the neurovascular bundle had been preserved. CONCLUSION: Preserving the neurovascular bundle on the side with a positive biopsy could result in a significantly greater incidence of PSM than with wide excision. Optimizing cancer control may require excision of the neurovascular bundle on a side known to have cancer on biopsy. In future site specific analyses, the PSM/ECE ratio could be used as a marker comparing cancer control outcomes from studies with differing technical approaches and indications for nerve-sparing surgery. PMID- 12581009 TI - Transrectal colour Doppler ultrasonography for quantifying angiogenesis in prostate cancer. AB - OBJECTIVE: To assess the correlation between angiogenesis and Doppler signal intensity using transrectal colour Doppler ultrasonography (CDUS) in patients with prostate cancer. PATIENTS AND METHODS: The study comprised 56 patients who underwent radical prostatectomy and had untreated tumours with a volume of> 0.1 mL in the peripheral zone. CDUS images were recorded on videotape before surgery. The Doppler signal intensity in tumours was evaluated using the colour pixel intensity (PI). Microvessel density (MVD) and vascular endothelial growth factor (VEGF) immunoreactivity were determined in the prostatectomy specimens. Microvessels were identified by immunohistochemical staining of endothelial cells for CD31. RESULTS: The PI in the tumour correlated with MVD (P < 0.001) and increased with higher levels of VEGF immunoreactivity (P = 0.004). There was no correlation between Gleason score and MVD or PI in the tumour. CONCLUSION: Blood flow assessed by CDUS may reflect the state of angiogenesis in prostate cancer. CDUS may be a useful technique for predicting tumour progression or prognosis, and may be useful for monitoring the effects of anti-angiogenic agents in the future. PMID- 12581010 TI - The clinical implications of the difference between castration, gonadotrophin releasing-hormone (GnRH) antagonists and agonist treatment on the morphology and expression of GnRH receptors in the rat ventral prostate. AB - OBJECTIVE: To examine the effects of castration and continuous gonadotrophin releasing-hormone (GnRH) agonist and antagonist therapy on the expression of GnRH receptors type I (GnRH-RI) in rat ventral prostate (VP) and pituitary tissue, and to compare the effects on prostate morphology. MATERIAL AND METHODS: Mature Sprague-Dawley rats were assigned to four treatment groups: group 1, vehicle only; group 2, GnRH agonist goserelin (100 microg/rat/day); group 3, GnRH antagonist cetrorelix (100 microg/rat/day); and group 4, orchidectomy. After 4 weeks the body weights were recorded and VP and pituitary tissue analysed for GnRH-RI expression using a competitive reverse transcriptase-polymerase chain reaction and immunohistochemistry. RESULTS: GnRH antagonist and orchidectomy decreased testosterone secretion and VP volume similarly, but the effects were not identical. The involution of the glandular lumina was more pronounced after orchidectomy while the antagonist therapy was more effective in suppressing epithelial cell proliferation. In the VP, GnRH-RI mRNA levels were increased after GnRH analogue therapy, but were unaffected by orchidectomy. In the pituitary, GnRH-RI mRNA expression was higher in response to orchidectomy and decreased after GnRH analogue treatment. CONCLUSION: Treatment with a GnRH antagonist for 4 weeks is more effective than an agonist in suppressing testosterone and inducing VP involution. The GnRH antagonist was more effective in suppressing VP epithelial cell proliferation than was castration, suggesting that it induces reduced proliferation by interfering with effects of locally produced GnRH. These results suggest that different regulatory mechanisms may operate in the rat VP than in the pituitary to control GnRH-RI mRNA expression. PMID- 12581011 TI - RhoA is associated with invasion and lymph node metastasis in upper urinary tract cancer. AB - OBJECTIVE: To assess the roles of RhoA small GTPase (RhoA) in upper urinary tract cancer by analysing the mRNA and protein levels of RhoA. PATIENTS AND METHODS: The mRNA and protein levels of RhoA in matched sets of tumour, non-tumour and metastatic lymph node tissues of surgical specimens were analysed in 47 consecutive patients with renal pelvic/ureteric cancer, using the polymerase chain reaction after reverse transcription and Western blotting. The relationship between mRNA and protein levels of RhoA in tumour tissues and the clinicopathological features of the patients was also assessed. RESULTS: The mRNA levels of RhoA and RhoA protein were greater in tumour and metastatic lymph node tissues than in non-tumour tissues (all P < 0.001). The expression levels of RhoA mRNA and protein levels in primary tumours was related to poorly differentiated grade (both P < 0.05) and muscle invasion (P < 0.01 and < 0.001, respectively). Kaplan-Meier plots of survival in patients with low or high RhoA showed that high mRNA and protein levels were associated with a shorter disease-free (P < 0.01) and overall survival (P < 0.001). Multivariate analysis using the Cox proportional hazards model showed that a high RhoA protein level was an independent prognostic factor, second to stage, in disease-free and overall survival (both P < 0.05). CONCLUSIONS: These findings suggest that RhoA is involved in the invasion and metastasis of upper urinary tract cancer, indicating that RhoA may be a useful prognostic factor in this disease. PMID- 12581013 TI - The urologist's guide to genital piercing. PMID- 12581012 TI - Standardization of the renogram technique for investigating the dilated upper urinary tract and assessing the results of surgery. AB - OBJECTIVE: To recommend a standard technique for the renographic evaluation of the dilated upper urinary tract, and for evaluating the results of surgery. METHODS: This review is an update of the previously recommended consensus report of the 9th International Symposium of Radionuclides in Nephrourology. RESULTS: State-of-the-art recommendations are detailed for standardizing common radionuclide investigations. CONCLUSIONS: Where open, laparoscopic or endourological techniques for treating pelvi-ureteric junction obstruction are under study, results can only be compared by standardizing the investigational techniques. The guidelines produced here are recommended for this purpose. PMID- 12581014 TI - Serum leptin levels in patients with premature ejaculation before and after citalopram treatment. AB - OBJECTIVE: To evaluate serum leptin levels (an adipocyte hormone involved in the suppression of appetite) in patients with premature ejaculation before and after treatment with citalopram, a selective serotonin reuptake inhibitor, with the hypothesis that leptin levels might become normal during this treatment. PATIENTS AND METHODS: The inhibitory effect of serotonin on libido, ejaculation and orgasm is well documented. Although there is no direct evidence of an association involving brain pathways which are related to sexual behaviour, there is an interaction between leptinergic and serotonergic systems. In a previous study serum leptin levels were high in patients with premature ejaculation. The present study comprised 30 patients with premature ejaculation according to the Diagnostic and Statistical Manual of Mental Disorders Third Revised Version. Fifteen patients (group I) were randomly assigned to 8 weeks of citalopram treatment and the remainder (15, group II) received no therapy. The patients were asked to determine the average intravaginal ejaculation latency time, and their fasting serum leptin levels were measured at baseline and after 8 weeks. RESULTS: There was no significant difference in the mean intravaginal ejaculation latency time between the groups at baseline; it increased after 8 weeks of treatment with citalopram in group I, to a mean (sd) of 209 (72.1) s, but not in group II. No difference was detected in leptin levels between the groups at baseline, but at 8 weeks they were lower in group I. CONCLUSION: As hypothesized, leptin levels decreased in patients with premature ejaculation after treatment with citalopram, and this decrease seemed to be linked to the therapeutic effect. Further experimental studies are needed. PMID- 12581015 TI - Estimation of glomerular filtration rate and bladder capacity: the effect of maturation, ageing, gender and size. AB - OBJECTIVE: To show that glomerular filtration rate (GFR) and bladder capacity (BC) develop isometrically, and to provide graphs for estimating GFR and BC for specific individuals of any age, sex, height and weight, for cystometric purposes. MATERIALS AND METHODS: GFR and BC data were collected from published reports; graphs relating GFR and BC to age, sex, height and weight were obtained by using a computerized curve-fitting technique that minimizes the 1.1 power of the absolute error. RESULTS: The plots show the GFR and BC for individuals of 10th, 50th and 90th percentile height and weight as a function of age and sex for different physiological conditions. GFR increases up to age 20 years and thereafter declines, whereas GFR per unit body surface area reaches a maximum at 3.25 years old and declines thereafter. CONCLUSIONS: The clinical management strategy to preserve or enhance renal function in paediatric and adult nephrological disease should incorporate the present data on development, growth, ageing and deterioration of function. These data should be used when interpreting cystometrograms and evaluating compliance. The graphs are useful for clinically estimating GFR and BC, especially when estimating infusion rate and BC for individual patients. BC and GFR develop isometrically with a proportionality constant of 4.56 min, except from birth to 1.5 years of age. Accordingly, individuals with healthy urinary systems, irrespective of age, sex and size, in the same physiological conditions have, on average, the same time to reach BC, ranging from 7 h at a normal 1% GFR to 41 min at the maximum diuresis of 10% GFR. PMID- 12581016 TI - A posterior-based omega-shaped flap vaginoplasty in girls with congenital adrenal hyperplasia caused by 21-hydroxylase deficiency. AB - OBJECTIVE: To describe a technical modification for constructing a vagina in girls with congenital adrenal hyperplasia caused by 21-hydroxylase deficiency, using a narrower skin base but rich subcutaneous tissue, aiming to obtain both longer and larger vaginal segments with better cosmesis of the external genitalia. PATIENTS AND METHODS: From August 1997 to February 2001, 28 girls (aged 5 months to 17 years) had a neovagina constructed using a posterior-based omega-shaped flap. Twenty-six patients had a low vagina entering into the urogenital sinus and two had a high vagina that entered the urogenital sinus. In those with a high vagina the flap procedure was combined with the Passerini Glazel technique. All the patients were scheduled for vaginal "calibrations" during the first year after surgery and, according to the result, would then undergo vaginal dilatation. RESULTS: Six children were re-operated; five had plastic surgery to correct genital folds that had regained a scrotal aspect, whereas one with a high vagina developed a urethral stricture, with urinary dribbling and infection, and had the urethra reconstructed. These six children are currently well. No hormone therapy was given to one child for 1 year who is scheduled for further surgery for a re-virilized clitoris. Two patients were lost to follow-up. Up to the last visit, 19 girls had not developed a vaginal stricture and the cosmesis of their external genitalia was deemed good. CONCLUSION: The posterior-based omega-shaped flap enabled both the construction of wider vaginal segments with a low risk of developing stenosis in those with a low vagina, and increased vaginal dimension when associated with the Passerini Glazel technique for those with a high vagina. However, despite good cosmesis of the external genitalia, the follow-up is too short to confirm whether this technique will meet all the expectations. PMID- 12581017 TI - Glanular reconstruction and preputioplasty repair for distal hypospadias: a unique day-case method to avoid urethral stenting and preserve the prepuce. AB - OBJECTIVE: To report the early outcome of >200 boys who underwent a glanular reconstruction and preputioplasty (GRAP) repair for distal hypospadias, to assess the functional and anatomical outcome of the penis at least 10 years after surgery, and to compare these data with otherwise "normal" aged-matched controls. PATIENTS AND METHODS: The GRAP repair is a novel method for the day-case reconstruction of distal hypospadias, and uniquely allows the anatomical reconstruction of the prepuce as part of penile reconstruction for hypospadias. We retrospectively reviewed the notes of 205 consecutive cases of distal hypospadias who had a GRAP repair carried out by one surgeon. Of these, there were 63 consecutive patients who had been repaired using the GRAP > or = 10 years earlier. In these patients a questionnaire was used to assess their perception of the appearance of their penis, the urinary stream, any problems with the foreskin and their recollection of surgery. Similar questionnaire data, except for the latter, were obtained from aged-matched controls for comparison. RESULTS: Of the 205 patients, 183 (89%) were day cases; the remaining 22 remained in hospital overnight for medical or social reasons. Sixteen (7%) developed a urethral fistula, the incidence of which correlated positively with the length of the repair. Four children (2%) were circumcised, in two for phimosis resulting in a spraying stream, in a third for dribbling after voiding, and the fourth for a cosmetically unsatisfactory foreskin. Questionnaire responses were received from 45 boys (71%), while 18 were untraceable. The mean (sd) age at surgery and at survey was 3.25 (2.6) and 14.26 (2.8) years, respectively. Forty-two (95%) children felt their penis was normal or only slightly different in appearance, two (5%) felt it looked fairly different and only one mostly avoided communal situations because of this. On voiding three boys (7%) sprayed most of the time while the remainder either never or sometimes sprayed. One youth sometimes had to sit to void because of difficulties in directing the stream; the rest (98%) always stood to void. There were no further circumcisions. These results were not significantly different from those of the age-matched controls. Most children had no (68%) or only slight (30%) recollection of hypospadias surgery. CONCLUSION: The complication rate and patient satisfaction with GRAP is comparable with those of other techniques. GRAP is a simple day-case procedure with few complications, thus avoiding an overnight hospital admission for most patients. Importantly, the prepuce can be preserved and refashioned to give a good cosmetic result, with no phimosis, which is increasingly important as circumcision becomes less acceptable to both the general public and the medical profession. PMID- 12581018 TI - Chemosensitization of human prostate cancer using antisense agents targeting the type 1 insulin-like growth factor receptor. AB - OBJECTIVE: To assess the effect of the downregulation of type 1 insulin-like growth factor receptor (IGF1R) on the chemosensitivity of prostate cancer cells. IGF1R is overexpressed by prostate cancer compared with benign prostatic epithelium and IGF1R expression commonly persists in androgen-independent metastatic disease at levels comparable to those in the primary. MATERIALS AND METHODS: Human androgen-independent DU145 prostate cancer cells were transfected with IGF1R antisense oligonucleotides or antisense RNA. Transfected cultures were treated with cisplatin, mitoxantrone, paclitaxel or vehicle control, and survival measured using a clonogenic assay. RESULTS: Both antisense strategies suppressed IGF1R protein levels to 30-50% of those in control cultures. This was associated with 1.5-2-fold enhancement of sensitivity to cisplatin, mitoxantrone and paclitaxel, and an increase in cisplatin-induced apoptosis. CONCLUSION: This approach has potential for development as a clinical treatment for advanced prostate cancer and other chemoresistant tumours. PMID- 12581019 TI - MUC1 expression, splice variant and short form transcription (MUC1/Z, MUC1/Y) in prostate cell lines and tissue. AB - OBJECTIVE: To detect the expression and transcription pattern of MUC1 in benign and malignant disease, and in two widely studied cell lines, and to investigate the glycosylation of MUC-1 in bone metastasis of prostate cancer, as mucins have been implicated in the progression and behaviour of several cancers. MATERIALS AND METHODS: RNA extracted from cell lines (DU145 and PC3), five samples of BPH and five samples of prostate cancer was reverse transcribed before amplification of MUC1-specific sequences by polymerase chain reaction. Paraffin-embedded sections were stained for glycosylated MUC1 and MUC1 core epitopes by HMFG1 and B27.29 antibodies, respectively. Steroid-treated cell lines were analysed by fluorescence-activated cell sorting, using the same antibodies. RESULTS: MUC1, in an under-glycosylated form, was widely expressed in the prostate and in metastatic lesions. MUC1/Z and MUC1/Y RNA were differentially expressed in BPH and prostate cancer, with no detectable expression of splice variant mRNA. This is in contrast to prostate cancer cell line cells (PC3 and DU145), which express splice variant mRNA. CONCLUSIONS: BPH, prostate cancer and metastatic prostate cancer all express high levels of under-glycosylated MUC1. This may explain the inability of previous studies to detect MUC1 in prostate tissue, as the antibody used was specific for a carbohydrate epitope which is not expressed on the under glycosylated MUC1. PMID- 12581022 TI - Comparison of percutaneous with transurethral cystolithotripsy in patients with large prostates and large vesical calculi undergoing simultaneous transurethral prostatectomy. PMID- 12581020 TI - In vivo evaluation of the potency and bladder-vascular selectivity of the ATP sensitive potassium channel openers (-)-cromakalim, ZD6169 and WAY-133537 in rats. AB - OBJECTIVE: To compare in vivo the potency and bladder-vascular selectivity of ATP sensitive potassium channel openers (KCOs) (-)-cromakalim, WAY-133537 and ZD6169 and a muscarinic antagonist, tolterodine in rats. MATERIALS AND METHODS: Bladder and arterial pressures were monitored simultaneously, before and after increasing intravenous doses of compounds, in each of two urethane-anaesthetized rat bladder hyperactivity models: spontaneous non-voiding myogenic contractions secondary to partial outlet obstruction and volume-induced neurogenic contractions. RESULTS: ( )-Cromakalim, WAY-133537 and ZD6169 caused a dose-dependent suppression of spontaneous contractions in the obstructed model, with a 50% inhibition of the contraction area under the curve at doses of 0.06, 0.14 and 2.4 micro mol/kg (intravenous), respectively. Corresponding decreases in mean arterial pressure at these effective doses were 24%, 15% and 15%, respectively. The KCO potency rank order was the same and their relative potency highly comparable in the neurogenic model. There was complete inhibition of spontaneous contractions in obstructed rats at doses corresponding to approximately 50% inhibition of the neurogenic contractions. While tolterodine caused a dose-dependent inhibition of contractions in the neurogenic model, it was ineffective at inhibiting non voiding contractions in obstructed rats. CONCLUSIONS: All KCOs tested caused significant decreases in arterial pressure at doses effective on the bladder in the model of obstructive instability, suggesting a lack of bladder-vascular selectivity. Similar KCO potency in both assays suggests no appreciable changes in KATP channel function as a result of partial outlet obstruction. PMID- 12581023 TI - Instillation of povidone iodine to treat and prevent lymphocele after renal transplantation. PMID- 12581024 TI - Multifocal transitional cell carcinoma in a patient with hereditary nonpolyposis colon cancer. PMID- 12581025 TI - Adenocarcinoma of the prostate immunostained for carbohydrate antigen 125 and carcinoembryonic antigen. PMID- 12581027 TI - Is microscopic haematuria a urological emergency? PMID- 12581028 TI - Penile fracture in Kermanshah, Iran: the long-term results of surgical treatment. PMID- 12581029 TI - End-to-end urethroplasty: long-term results. PMID- 12581030 TI - Nocturia in relation to sleep, somatic diseases and medical treatment in the elderly. PMID- 12581034 TI - Satisfaction with team midwifery care for low- and high-risk women: a randomized controlled trial. AB - BACKGROUND: In 1996 a new model of maternity care characterized by continuity of midwifery care from early pregnancy through the postpartum period was implemented for women attending Monash Medical Centre, a tertiary level obstetric service, in Melbourne, Australia. This study's purpose was to assess the impact of this model on women's views and experiences of care during the antenatal, intrapartum, and postpartum periods compared with views of women receiving standard maternity care. METHODS: One thousand low- and high-risk women who booked at the antenatal clinic and met the eligibility criteria were randomly allocated to continuity of midwifery care from a group of seven midwives in collaboration with medical staff, or to standard care from a variety of midwives and medical staff. Women's views of care were measured by means of a postal questionnaire at four months after the birth. RESULTS: Team midwifery care was associated with increased satisfaction with antenatal, intrapartum, and some aspects of postpartum care. The differences were most obvious for antenatal care. CONCLUSIONS: Continuity of midwifery care is realistically achievable in a tertiary obstetric referral service and is associated with increased satisfaction. PMID- 12581035 TI - Swedish women's interest in home birth and in-hospital birth center care. AB - BACKGROUND: In Sweden, few alternatives to a hospital birth are available, and little is known about consumer interest in alternative birth care. The aim of this study was to determine women's interest in home birth and in-hospital birth center care in Sweden, and to describe the characteristics of these women. METHODS: All Swedish-speaking women booked for antenatal care during 3 weeks during 1 year were invited to participate in the study. Three questionnaires, completed after the first booking visit in early pregnancy, at 2 months, and 1 year after the birth, asked about the women's interest in two alternative birth options and a wide range of possible explanatory variables. RESULTS: Consent to participate in the study was given by 3283 women (71% of all women eligible). The rates of response to the three questionnaires were 94, 88, and 88 percent, respectively. One percent of participants consistently expressed an interest in home birth on all three occasions, and 8 percent expressed an interest in birth center care. A regression analysis showed five factors that were associated with an interest in home birth: a wish to have the baby's siblings (OR 20.2; 95% CI 6.2-66.5) and a female friend (OR 15.2; 95% CI 6.2-37.4) present at the birth, not wanting pharmacological pain relief during labor and birth (OR 4.7; 95% CI 1.4-15.3), low level of education (OR 4.5; 95% CI 1.8-11.4), and dissatisfaction with medical aspects of intrapartum care (OR 3.6; 95% CI 1.4-9.2). An interest in birth center care was associated with experience of being in control during labor and birth (OR 8.3; 95% CI 3.2-21.6), not wanting pharmacological pain relief (OR 2.3; 95% CI 1.3-4.1), and a preference to have a known midwife at the birth (OR 2.2; 95% CI 1.6-2.9). CONCLUSION: If Swedish women were offered free choice of place of birth, the home birth rate would be 10 times higher, and the 20 largest hospitals would need to have a birth center. Women interested in alternative models of care view childbirth as a social and natural event, and their needs should be considered. PMID- 12581036 TI - Satisfaction and use of prenatal care: their relationship among African-American women in a large managed care organization. AB - BACKGROUND: Although many more mothers of almost all ethnic groups began prenatal care in the first trimester during the last decade, a significant number of low income and minority women still fail to obtain adequate care in the United States a failure that may be related to their dissatisfaction with the prenatal care experience. This study sought to examine the relationship between satisfaction with care and subsequent prenatal care utilization among African-American women using prospective methods. METHODS: A sample of 125 Medicaid and 275 non-Medicaid African-American adult women seeking care through a large Midwest managed care organization were interviewed before or at 28 weeks' gestation at one of two prenatal care sites. Women were interviewed about personal characteristics, prenatal care experience, and ratings of care (satisfaction). Information about subsequent use of prenatal care was obtained through retrospective medical record review after delivery. Univariate and multivariable analyses examining the relationship between women's satisfaction and prenatal care use were conducted using a dichotomous measure of satisfaction and a continuous measure of utilization. RESULTS: Women were highly satisfied with prenatal care, with an overall mean satisfaction score of 80.3. Non-Medicaid women were significantly (p < 0.05) less satisfied with their prenatal care (mean score, 79.1) than Medicaid women (mean score, 82.8), and the latter had significantly fewer visits on average than the former subsequent to the interview. Analyses showed no significant difference in subsequent utilization according to whether a woman had a high versus low level of satisfaction at the prenatal care interview. CONCLUSIONS: This study challenges the assumption that improving a woman's satisfaction with care will lead to an increase in the adequacy of her prenatal care utilization. Since this study was limited to African-American women and is the first prospective study of women's satisfaction with care and prenatal care utilization, the negative findings do not yet settle this area of inquiry. Monitoring women's satisfaction with prenatal care in both managed care and fee for-service settings and working to improve those aspects of care associated with decreased satisfaction is warranted. PMID- 12581037 TI - Parent visiting and participation in infant caregiving activities in a neonatal unit. AB - BACKGROUND: Active parent involvement in caring for their infants in the neonatal care unit is thought to improve parent-infant attachment and to moderate the psychological stress for parents, but few recent studies have examined parent visiting patterns and participation in infant caregiving. The study purposes were to describe the frequency and duration of parent visiting and participation in infant caregiving activities, and to identify parent and infant factors associated with parental participation. METHODS: Parental visiting frequency, duration, and participation in social, cleaning, and feeding activities with their infant (n=110) were recorded on 12 days during a 3-month period in a tertiary neonatal unit. RESULTS: Mothers visited more frequently (85% vs 45% of possible days) and for longer than fathers, and visited less frequently if the infant had other siblings, if the infant was over age 1 month, or if fathers made fewer visits. Fathers visited less frequently if the infant was over age 7 days and more frequently if the mothers visited more frequently. All mothers and most (96%) fathers carried out social activities, such as talking, stroking or holding, during their visits. Over 75 percent of mothers engaged in infant cleaning and feeding activities during visits in contrast with less than 20 percent of fathers. Mothers' participation in infant feeding was best predicted by the duration of their visit and their participation in infant cleaning. Fathers' participation in infant feeding was only related to their participation in infant cleaning. CONCLUSIONS: Significant differences were found in this neonatal unit between mothers' and fathers' visiting patterns and infant caregiving activities. Neonatal unit staff should consider factors that may influence parental visiting and explore strategies to improve parental involvement in caregiving. PMID- 12581038 TI - Prevalence and predictors of women's experience of psychological trauma during childbirth. AB - BACKGROUND: The increased acceptance of the prevalence of trauma in human experience as well as its psychological consequences has led to revisions of diagnostic criteria for the disorder. The three purposes of this study were to examine the rates at which women experienced psychological trauma in childbirth, to explore possible causal factors, and to examine possible factors in the development of the disorder. METHODS: One hundred and three women from childbirth education classes in the Atlanta metropolitan area completed a survey in late pregnancy and a follow-up interview approximately 4 weeks after the birth. RESULTS: The childbirth experience was reported as traumatic by 34 percent of participants. Two women (1.9%) developed all the symptoms needed to diagnose posttraumatic stress disorder, and 31 women (30.1%) were partially symptomatic. Regression analysis showed that antecedent factors (e.g., history of sexual trauma and social support) and event characteristics (e.g., pain in first stage of labor, feelings of powerlessness, expectations, medical intervention, and interaction with medical personnel) were significant predictors of perceptions of the childbirth as traumatic. The pain experienced during the birth, levels of social support, self-efficacy, internal locus of control, trait anxiety, and coping were significant predictors of the development of posttraumatic stress disorder symptoms after the birth. CONCLUSIONS: These findings suggest several intervention points for health care practitioners, including careful prenatal screening of past trauma history, social support, and expectations about the birth; improved communication and pain management during the birth; and opportunities to discuss the birth postpartum. PMID- 12581039 TI - "Vanished twin": an exploration of women's experiences. AB - BACKGROUND: Evidence suggests that in identified twin pregnancies 30 percent become a "vanished twin," that is, when two gestation sacs are identified at early ultrasound but subsequently only one fetus is seen. This study explored the thoughts, feelings, and hospital experiences of women who have a vanished twin. METHOD: The study was conducted in a large teaching hospital in northwest England from April to June 2001. The experiences of 9 women were explored through 4 focus groups and an individual interview. Nulliparous and multiparous groups were separated for homogeneity. The study's philosophical approach was based on constructivism, and the thematic analysis used a feminist interpretation to search for patterns of meanings from the women's words. RESULTS: The women wanted to be cared for in a humanistic manner, and they felt that acknowledgement of the vanished twin by the professional was important. The findings highlighted the need for better information giving to improve communication between caregivers and women. Multiparas were more prepared than nulliparas to ask for the attention that they wanted. CONCLUSIONS: Despite the small number of study participants, results suggest that maternity caregivers should be sensitive to the feelings and individual nature of a woman who has experienced a vanished twin. Humanistic approaches to communication and improved methods of information based on the development of an information sheet are advised. PMID- 12581040 TI - Sheila Kitzinger's letter from Europe: the clock, the bed, the chair, the pool. PMID- 12581041 TI - Home birth versus hospital birth: questioning the quality of the evidence on safety. PMID- 12581043 TI - Term Breech Trial. PMID- 12581045 TI - Term Breech Trial. PMID- 12581046 TI - Role of supervision in a radiology practice. PMID- 12581047 TI - Frontal recess air cells: spectrum of CT appearances. AB - The CT appearances of frontal recess air cells is reviewed. PMID- 12581048 TI - MRI in ankle tuberculosis: Review of 14 cases. AB - We reviewed the MR imaging features of ankle tuberculosis and determined the role of MR in its diagnosis. A retrospective analysis of 14 cases of ankle tuberculosis imaged with MRI was performed. Plain radiographs were also reviewed where available, and the imaging characteristics were noted. We also reviewed the medical records in order to assess the impact of the imaging findings on management of these patients. Magnetic resonance imaging is extremely helpful for detection, mapping the extent and resolution of the disease. It can identify cases, enables early institution of antituberculous chemotherapy and might obviate the need for surgery. PMID- 12581049 TI - Pancreatic cancer: preliminary experience with sodium iodide fluorodeoxyglucose positron emission tomography in Australia. AB - Previous studies of fluorodeoxyglucose positron emission tomography (FDG-PET) in pancreatic cancer have used Bismuth Germinate detector systems. This preliminary Australian study aims to confirm the accuracy of FDG-PET in pancreatic cancer using a dedicated sodium iodide (NaI) PET system. Fifteen consecutive patients underwent FDG-PET using a GE QUEST dedicated NaI PET scanner. The indications were the characterization of a pancreatic mass seen on CT or ultrasonographic imaging (nine cases), diagnosis or exclusion of recurrent disease following surgery and adjuvant therapy (four cases) and presurgical staging of primary pancreatic cancer (two cases). The final diagnosis was determined from histology or, when no histology was available, by radiological and clinical follow up. The FDG-PET accurately characterized eight out of nine pancreatic masses (seven were true negative, one was true positive and one was false positive). Of the four cases performed to determine recurrent disease, three were accurately diagnosed (two true negatives and one true positive). In the fourth case, PET accurately detected a liver metastasis but did not detect the local recurrence. Results in the two cases where PET was performed for preoperative staging comprised one true positive and one false negative. Sodium iodide FDG-PET is useful in the diagnosis of pancreatic cancer, particularly in the presence of a previously detected mass. PMID- 12581050 TI - Renal colic: a prospective evaluation of non-enhanced spiral CT versus intravenous pyelography. AB - The aim of this study was to compare non-enhanced spiral CT (NECT) and intravenous pyelography (IVP) in patients with suspected acute renal colic. Two hundred patients presenting to the Emergency Department with suspected acute renal colic were randomized into groups undergoing NECT or IVP. The main outcome measures were diagnostic utility, incidence of alternative diagnoses, requirement for further imaging, length of hospital stay, urological intervention rates, radiation dosage and costs. Non-enhanced spiral CT was better than IVP in making a definitive diagnosis of ureteric calculus or of recent calculus passage (65/102 or 66% vs 42/98 or 41%; P = 0.003). Calculi were missed in two patients in the IVP group. Two patients in each group had alternative diagnoses by initial imaging. There was no difference in the length of hospital stay or intervention rate. More plain X-rays during admission and more IVPs during follow up were performed in the NECT group. Effective radiation dosages were 2.97 mSv (IVP) and up to 5 mSv (NECT). Non-enhanced spiral CT provided greater diagnostic utility in this randomized comparison but no difference in measured outcomes. The incidence of alternative diagnoses was low, probably due to patient selection. Financial costs for each modality are comparable in a public tertiary hospital. Radiation dosages are higher for NECT and, for this reason, it might be appropriate to consider limiting NECT use to patients who have do not have classical symptoms of renal colic, to older patients and those with a contraindication to the administration of intravenous contrast media. PMID- 12581051 TI - Solitary hemispheric demyelination in acute disseminated encephalomyelitis: clinicoradiological correlation. AB - We retrospectively studied 13 patients with solitary hemispheric demyelination in acute disseminated encephalo--myelitis (solitary-ADEM) to look for specific MRI features. Thirteen patients were subjected to routine MRI with varying initial clinical diagnosis, including demyelination, neoplasm, encephalitis and infarct. The provisional MRI diagnosis was based on lesion morphology, size, location, mass effect, effect on adjacent sulci, cisterns and gyral shape. On long repetition time (TR)/long echo time (TE) spin echo sequences, the lesions were heterogeneously hyperintense, and on short TR/short TE spin echo sequences, heterogeneously hypointense. Later, the provisional MRI diagnosis was corroborated with clinical parameters such as multimode-evoked potentials, electro--physiological studies and analysis of cerebrospinal fluid (CSF) in order to establish a definitive diagnosis. The diagnosis was established in all except one patient with tumour-like clinical and MRI features. The combination of MRI features and clinical parameters can establish a definitive diagnosis in the majority of cases, and this avoids a biopsy. PMID- 12581052 TI - Can virtual simulation of breast tangential portals accurately predict lung and heart volumes? AB - A treatment portal or simulator image has traditionally been used to demonstrate the lung and heart coverage of the breast tangential portal. In many cases, these images were acquired as a planning session on the linear accelerator. The patients were also CT scanned to assess the lung/heart volume and to determine the surgical site depth for the electron-boost energy. A study using 50 consecutive patients was performed comparing the digitally reconstructed radiograph (DRR) from the virtual simulation with treatment portal images. Modification to the patient's arm position is required when performing the planning CT scans due to the aperture size of the CT scanner. Virtual simulation was used to assess the potential variation of lung and heart measurements. The average difference in lung volume between the DRR and portal image was less than 2 mm, with a range of 0-5 mm. Arm position did not have a significant impact on field deviation; however, great care was taken to minimize any changes in arm position. The modification of the arm position for CT scanning did not lead to significant variations between the DRRs and portal images. The Advantage Sim software has proven capable of producing good quality DRR images, providing a realistic representation of the lung and heart volume included in the treatment portal. PMID- 12581053 TI - Why a breast boost should remain a controversial aspect of routine breast conservation management in Australia and New Zealand in 2002. AB - Two randomized trials have recently shown a statistically significant improvement in local control when a boost is employed in the conservative treatment of early breast cancer. However, unresolved issues of cost effectiveness, potentially increased toxicity and the inability to automatically generalize these results to Australian and New Zealand practice remain. In view of these unresolved controversies, the St George and Wollongong hospitals breast boost trial (SGW trial) will continue to recruit. PMID- 12581054 TI - Oncology service correspondence: do we communicate? AB - The aim of this study is to assess the content of letters sent from a hospital oncology service regarding the initial assessment of new referrals and the requirements of hospital specialists and General Practitioners (GPs) regarding oncology service correspondence. The content of letters regarding the initial consultation of 204 consecutive new oncology patients was analysed. General Practitioners and referring specialists were sent a 13-point survey to gauge their preferences for the information contained in oncology correspondence pertaining to the initial assessment of an oncology patient. Seventy-two percent of the patients had a letter written following their initial oncology assessment. The GP received a copy of 81% of the letters (58% of the study sample). The diagnosis was recorded in all our letters, and the proposed treatment plan was addressed in 84% of our letters. Both GPs and specialists required information on examination and investigation findings, diagnosis, treatment options, proposed management plan, and what the patient was told. The GPs required further information on current medication, likely side-effects of the proposed management, and clarification of when to contact the oncologist. The majority of the respondents were in favour of a structured letter. PMID- 12581055 TI - MRI in kernicterus. AB - Kernicterus (bilirubin encephalopathy) is a disease entity with a very high rate of mortality in neonates. The children who survive are left with neurological deficits such as choreoathetosis, sensorineural deafness and mental retardation. Magnetic resonance imaging of the brain in this condition has specific findings which aid in the accurate diagnosis of the condition, along with clinical and biochemical criteria. Reports have shown involvement of the globus pallidus, putamen, and less commonly the thalamus. More specific involvement may be isolated lesions of the globus pallidus. Magnetic resonance imaging has shown a good sensitivity in this condition. PMID- 12581056 TI - Coexistent gallbladder carcinoma in Mirizzi syndrome. AB - A 52-year-old woman presented with right upper quadrant pain and obstructive jaundice. Computed tomographic scan showed Mirizzi syndrome type 1 and a thickened and calcified gallbladder wall, raising the possibility of coexistent gallbladder carcinoma that was later confirmed on histology post-cholecystectomy. PMID- 12581057 TI - Murray Valley encephalitis: case report and review of neuroradiological features. AB - We report on a child with diffuse symmetrical thalamic enlargement and signal increase on MRI, representing changes caused by Murray Valley encephalitis (MVE). Very little has previously been reported on the neuroradiological findings of MVE, also known as Australian encephalitis. It is endemic to tropical North Australia, particularly Western Australia and the Northern Territory, but can occur in other parts of Australia. The last epidemic was in south-eastern Australia in 1974. Australian encephalitis is the second most serious acute viral encephalitis to be encountered in Australia. Clinicians need to be aware of MVE in this era of ever-increasing travel. Our aim is to highlight these finding and further define the neuroradiological features. PMID- 12581058 TI - Thrombin injection for repair of pseudoaneurysms: a case for caution. AB - Treatment of pseudoaneurysms by the injection of thrombin have been reported in the literature with a success rate approaching 100%. Complications have been reported with its use. We report a case where thrombin was used to thrombose a superficial temporal artery pseudoaneurysm leading to the development of seizure and ischaemia of the scalp. We advise that thrombin should be not be used to thrombose pseudoaneurysms of arteries supplying critical areas. PMID- 12581059 TI - Detection of aortoenteric fistula with helical CT. AB - We describe a case of a patient with symptoms suggestive of secondary aortoenteric fistula (AEF). Extravasation of contrast medium from the aorta into the lumen of the small bowel was definitive evidence of AEF. The role of abdominal CT performed without oral contrast is discussed. PMID- 12581061 TI - Abnormal uptake of Technetium-99 m hydroxymethylene diphosphonate in the liver. AB - Diffuse liver uptake on bone scintigraphy is a rare occurrence with only a few reports in the literature. A case of diffusely increased uptake in a patient with acute hepatitis is presented. PMID- 12581060 TI - Imaging of primary leiomyosarcoma of the seminal vesicle. AB - Leiomyosarcoma of the seminal vesicle is exceedingly rare. The ultrasound and CT findings in a recent case are described. These consist of a mass causing enlargement of the right seminal vesicle with involvement of the urinary bladder and prostate. The prostate gland was displaced inferiorly and to the left. Surgical resection and subsequent histology confirmed the diagnosis. The role of imaging in retrovesical masses is also discussed. PMID- 12581062 TI - Intracranial Tc99m MDP uptake in Sturge-Weber syndrome. AB - A case of intracranial extraosseous 99mTc MDP uptake is presented, which was found on CT to be due to dystrophic gyral calcification characteristic of Sturge Weber syndrome (SWS). The imaging characteristics of SWS are described. The possibility of extraosseous MDP uptake should be considered when unusual or atypical 'hot spots' are seen on bone scanning. PMID- 12581063 TI - Mesenchymal hamartoma of the chest wall in infants: report of three cases and literature review. AB - Mesenchymal hamartoma of the chest wall in infants is a rare abnormality. We present a report of three cases and a brief review of the literature, emphasizing the role of the radiologist in diagnosis and management. PMID- 12581064 TI - Midgut volvulus in an adult. AB - We report a case of duodenal malrotation that presented in adulthood as midgut volvulus. On colour Doppler ultrasound, findings of a clockwise rotation of the superior mesenteric vein around the superior mesenteric artery (whirlpool sign) and a distinctive 'whirl'-like pattern resulting from an encircling of loops of the bowel around the SMA on CT is diagnostic of midgut volvulus. PMID- 12581065 TI - MRI in greater trochanter pain syndrome. AB - The greater trochanter pain syndrome refers to pain on the lateral aspect of the hip joint. This is frequently attributed to trochanteric bursitis and distension of the subgluteal bursae. Associated tears of the tendons of gluteus medius and minimus have been described and may result from repetitive frictional trauma to these tendons and their associated bursae secondary to impingement beneath the tensor fascia lata. Occasionally tendinous damage may result from acute local direct trauma or a hyperadductive strain injury. We describe MRI in two patients with chronic lateral hip pain. PMID- 12581067 TI - MRI of partial anomalous pulmonary venous return (scimitar syndrome). AB - We report a case of anomalous pulmonary venous drainage into the inferior vena cava (scimitar syndrome). Cine MRI and 3-D contrast-enhanced MR angiography provides an non-invasive diagnostic technique in the evaluation of anomalous pulmonary venous return. PMID- 12581066 TI - Primary parenchymal cerebral cystic haemangiopericytoma: a 5-year follow up of disease progression. AB - Haemangiopericytoma (HPC) is a rare tumour of the central nervous system. Previously, HPCs were thought to originate from meninges and ventricular walls. Currently, they are accepted as distinctive mesenchymal neoplasms unrelated to meningiomas. Imaging appearances, clinical progression and haemorrhage into a cystic tumour is documented in an 18-year-old man where the final diagnosis, with histopathological verification, proved to be a cystic HPC. This interesting and illustrative case is discussed with the relevant literature. PMID- 12581068 TI - Aneurysms of the inferior vena cava. AB - Two cases of saccular aneurysms of the infrarenal inferior vena cava (IVC) associated with retrohepatic IVC obstruction are described. Ultrasonographic, computerized tomographic and inferior venacavography findings in these cases are presented. PMID- 12581069 TI - Re: Angiography-related transient cortical blindness in patients with adult polycystic kidney disease. PMID- 12581076 TI - Methods of evaluating hair growth. AB - For decades, scientists and clinicians have examined methods of measuring scalp hair growth. With the development of drugs that stem or even reverse the miniaturization of androgenetic alopecia, there has been a greater need for reliable, economical and minimally invasive means of measuring hair growth and, specifically, response to therapy. We review the various methods of measurement described to date, their limitations and value to the clinician. In our opinion, the potential of computer-assisted technology in this field is yet to be maximized and the currently available tools are less than ideal. The most valuable means of measurement at the present time are global photography and phototrichogram-based techniques (with digital image analysis) such as the 'TrichoScan'. Subjective scoring systems are also of value in the overall assessment of response to therapy and these are under-utilized and merit further refinement. PMID- 12581077 TI - Imaging the skin. AB - Since the discovery of X-rays, the use of imaging technology has continued to play an important role in medicine. Technological advancements have led to the development of various imaging modalities, most of which have been used to image organs deep within the human body. More recently, attention has focused on the application of imaging technology for evaluation of the skin. A variety of techniques are currently being used to examine the skin and these include specialized photography, surface microscopy, ultrasound, laser Doppler perfusion imaging, confocal microscopy, and magnetic resonance imaging. These modalities can provide information that can assist in the management of skin problems. Although many of these techniques are still undergoing research, they are showing promise as useful clinical tools in dermatology. PMID- 12581078 TI - Paediatric atopic dermatitis in Perth general practice. AB - Ninety-seven Perth general practitioners completed a self-administered postal questionnaire that aimed to examine their caseload and management practices for childhood atopic dermatitis (AD). General practitioners saw a median of two new cases and three follow-up consultations per month for childhood AD, and referred a median of 10% of cases to a specialist, usually a dermatologist. Most (77%) recommended emollients for all patients, but only 21% specifically reported advising their use immediately after bathing. Sixty-one percent would use topical corticosteroids in all or most patients, but cream preparations were more commonly used (58%) than ointments (40%). Atrophy was rated as a common or very common side-effect of topical corticosteroid therapy by 23% of general practitioners. Twenty-six percent reported using oral corticosteroids in children with AD. Dietary changes would be recommended in at least a few AD patients by 79% of general practitioners, and 31% would recommend a change from cow's milk to soy in the absence of a history of dietary triggers. We conclude that general practitioners appeared generally well informed about AD management. However, dermatologists, through targeted education, may be in a position to help general practitioners further improve outcomes for these patients. PMID- 12581079 TI - Solar keratoses: analysis in a dermatological practice in Australia. AB - Two hundred consecutive patients with solar keratoses (SK) seen in a private dermatology practice had on average 61.9 SK compared with eight reported in the general population. Non-melanoma skin cancer was present in 41% of patients and 17% had squamous cell carcinoma (SCC). The ratio of basal cell carcinoma to SCC in the cohort was 1.7:1. The commonest site of SK was the upper limbs but the greatest density of lesions was on the face, particularly the nose. Squamous cell carcinomas were most commonly found on the upper and lower limbs. Basal cell carcinomas were most common on the head and neck. PMID- 12581080 TI - Topical diclofenac in hyaluronan gel for the treatment of solar keratoses. AB - This randomized, double-blind, placebo-controlled study assessed the efficacy and safety of a topical gel containing 3% diclofenac in 2.5% hyaluronan in 150 patients with solar keratoses (SK). The active treatment was compared with the vehicle only, hyaluronan gel, as placebo over a 12-week period. Patients in both groups applied the active treatment or placebo to a targeted area of skin (0.25 g b.d.). At 12 weeks the mean lesion-count reduction in the targeted area was not significantly different between treatments. However, at post-termination follow up (16 weeks), there was a highly significant decrease in the number of lesions, 6.2 +/- 7.5 standard deviations (SD) (56.1% reduction) in the active treatment group compared with 2.4 +/- 4.3 SD (23.6% reduction) in the placebo group (P < 0.001). Other efficacy measures (complete lesion resolution, >50% lesion reduction) were also significantly different (P < 0.01) between treatments at 16 weeks. In conclusion, topical 3% diclofenac in 2.5% hyaluronan gel was effective and well tolerated in this study, suggesting a role for this therapy in the treatment of SK. PMID- 12581081 TI - Eosinophilic pustular folliculitis (Ofuji's disease) in Singapore: a review of 23 adult cases. AB - The case records of 23 patients with classic eosinophilic pustular folliculitis (EPF), or Ofuji's disease, seen at the National Skin Centre in Singapore, from 1990 to 2001 were reviewed. All patients had clinical and histopathological findings consistent with EPF. There were eight men and 15 women (ratio 1:1.6). The mean age at presentation was 35 years. There was a marked predilection for Chinese patients (87%), with a racial distribution of 20:2:1 of Chinese, Malay and Indian patients, respectively. The most frequent site of occurrence was the face, particularly over both cheeks. The majority of patients (90%) treated with oral indomethacin had a good response within 2-4 weeks. Relapses were frequent in 82.6% of patients and maintenance with indomethacin or ketoprofen was beneficial. Eosinophilic pustular folliculitis is a rare but important disease entity presenting with recurrent indurated erythematous papulopustules and plaques on the face. Increased awareness of this condition is important as it can mimic many other conditions presenting as red plaques on the face. PMID- 12581082 TI - Scurvy: a cutaneous clinical diagnosis. AB - Two cases of scurvy diagnosed following presentation with a purpuric rash are presented. A 44-year-old man developed scurvy as a result of poor dietary intake of vitamin C. This occurred because of a number of factors. including poor dentition, diarrhoea, depression and benzodiazepine/narcotic dependence. A 69 year-old man with acute myeloid leukaemic transformation of myelodysplastic syndrome developed mucositis, nausea, vomiting and diarrhoea as complications of chemotherapy. This led to poor dietary intake and consequently scurvy. Both cases demonstrated specific and diagnostic cutaneous manifestations of scurvy, particularly perifollicular purpura, ecchymoses and coiled corkscrew hairs. The diagnosis was supported by specific diet history. Ascorbic acid tolerance test was used as a simple laboratory method to confirm the clinical diagnosis. PMID- 12581083 TI - Birt-Hogg-Dube syndrome. AB - We present three members of a Queensland family with clinical and histopathological features consistent with Birt-Hogg-Dube syndrome. Two of the three family members were able to be screened for associated disorders. The mother of the family was found to have a solitary colonic polyp, a large ovarian cyst and two chorioretinal scars. No associated disorders were found on investigation of one of the two affected sons. PMID- 12581084 TI - Warfarin-induced skin necrosis associated with acquired protein C deficiency. AB - A 36-year-old woman developed skin necrosis of the inner thighs following the re introduction of warfarin after a laparoscopic cholecystectomy. She had a history of liver disease and cardiomyopathy and was on warfarin for 10 years. Warfarin induced skin necrosis secondary to protein C deficiency was diagnosed. Although warfarin was ceased immediately, the prothrombin time measurements remained prolonged and warfarin levels remained therapeutic. Our patient, who had attached great significance to warfarin therapy, had continued the ingestion of warfarin despite our advice. She required three surgical debridements. Protein C levels, as measured 1 year later, were within normal limits, confirming the transient nature of the acquired deficiency during the acute event. This is the second reported case of warfarin necrosis associated with acquired protein C deficiency. PMID- 12581085 TI - Androgenetic alopecia in a postmenopausal woman as a result of ovarian hyperthecosis. AB - A 65-year-old woman presented with an 8-year history of progressive frontotemporal alopecia and hirsutism. She had elevated serum levels of testosterone, androstenedione and estradiol. Ultrasound and computed tomography imaging suggested a right ovarian mass, while bilateral ovarian venous sampling demonstrated increased testosterone levels originating from both ovarian veins. Histology obtained following bilateral oophorectomy demonstrated bilateral ovarian hyperthecosis. Six months after surgery, the patient remains well with no progression of the alopecia. Ovarian hyperthecosis is a rare cause of androgenetic alopecia in postmenopausal women. The role of hyperthecosis and its relationship to androgenetic alopecia is reviewed. PMID- 12581086 TI - O'Brien's actinic granuloma in association with prolonged doxycycline phototoxicity. AB - O'Brien's actinic granuloma is clinically characterized by annular papules and plaques on sun-exposed areas of skin. These lesions often occur insidiously on a background of severe solar elastosis; however, an acute variant following sunburn has been reported in the literature. We present two cases of acute actinic granuloma precipitated by episodes of sunburn occurring on a background of prolonged doxycycline phototoxicity. Biopsies from both patients showed a histiocytic infiltrate with multinucleate giant cells engulfing elastotic material, with a reduction of elastin towards the centre of the papule. Marked resolution of the lesions was noted after 8 weeks of treatment with betamethasone dipropionate 0.05% ointment in optimized vehicle together with adequate photoprotection in the form of broad-spectrum sunscreens. PMID- 12581087 TI - Twenty-nail dystrophy in a girl with incontinentia pigmenti. AB - A 9-year-old girl with incontinentia pigmenti (IP) diagnosed soon after birth subsequently developed 20-nail dystrophy at the age of 3 years. This persists unchanged 6 years later, and she has no other evidence of any other cutaneous disease, nor a family history of IP or nail dystrophy. Twenty-nail dystrophy has been associated with a number of diseases including eczema, lichen planus, alopecia areata and psoriasis and a number of other congenital associations, but the association with IP appears to be unique. PMID- 12581088 TI - Ulcerated juvenile xanthogranuloma of the scalp. AB - A 7-month-old girl presented with a single rapidly enlarging thickly crusted lesion on her frontal scalp. Histology was consistent with a juvenile xanthogranuloma (JXG) with ulceration and surface crust formation. Reports of ulcerating, crusted lesions of JXG are rare and the few reports have been confined to giant forms or those involving mucosal sites. This unusual presentation delayed diagnosis. Our report highlights the great variability in clinical presentation of JXG and is a reminder to consider this diagnosis in the differential of any lesion developing in an infant, particularly on the head, neck and upper trunk. PMID- 12581089 TI - Mondor's disease and electrocution. PMID- 12581090 TI - Narrow-band ultraviolet B phototherapy for acquired perforating dermatosis. PMID- 12581091 TI - Epidemiology and pathogenesis of scleroderma. AB - Scleroderma is a heterogenous connective tissue disorder characterized by fibrosis of the skin, with or without internal organ involvement. The aetiology of scleroderma may involve both environmental and genetic factors. Abnormalities involving the immune system, vascular tissue and extracellular matrix have been demonstrated. Recent research has focused on microchimerism as a risk factor for the development of scleroderma. This article reviews the epidemiology and pathogenesis of this disorder. PMID- 12581097 TI - Unfulfilled expectations and negative interactions: nonsupport in the relationships of women caregivers. AB - BACKGROUND: Most research has viewed social support as unconditionally positive, but negative interactions such as conflict and dissatisfaction with support are present in caregivers' relationships and have a negative impact on their health. PURPOSE: The purpose of this study was to describe women's experience of nonsupport and identify the types of nonsupport present in their relationships with family and friends while caring for a family member with dementia. ETHICS: The study received ethical clearance from the university ethics review committee. METHODS: The theoretical perspective of symbolic interaction guided this longitudinal qualitative research. Included in the study were 25 in-depth interviews with a group of eight women selected from a larger study of family caregiving because they described nonsupport in their relationships with family and friends. FINDINGS: The women experienced two types of problems in realizing support. Unmet expectations for support was the most pervasive difficulty and included unfulfilled or missing offers of assistance, unmet expectations for social interaction, mismatched aid, or incompetence on the part of the potential helper. Negative interactions, particularly with kin, took several forms: disparaging comments that belittled a caregiver's experience, conflict between the caregiver and others in their appraisal of the care recipient's health status, criticism of the caregiver's decisions in caring for the care recipient, and spillover from longstanding conflict and issues in the family history. LIMITATIONS: This study was limited to women caring for an adult relative with dementia and included a relatively homogeneous sample of English speaking, North American women of European descent. CONCLUSIONS: This study highlighted the utility of identity theory to explain the impact of unmet expectations for support and recognized disparaging comments as a form of nonsupportive interactions. PMID- 12581098 TI - Can burdened caregivers be effective facilitators of elder care-recipient health care? AB - BACKGROUND: Caregiving by informal family caregivers of dependent older people in the community may be affected by burden and by the personal and social resources available to the caregiver. Given the increase in the dependent older population, study of factors affecting informal caregiving is necessary. AIMS: To examine caregiver resources, burden and competence as predictors of health-care facilitation on behalf of older patients. DESIGN: Cross-sectional data were collected by an interview schedule from 240 randomly sampled spousal and filial caregivers in Jerusalem. Study variables included caregiver background variables, general self-concept and feelings of caregiver competence, informal and formal social support, burden and levels of health-care facilitation. Path analysis was performed to clarify the direct and indirect predictors of health-care facilitation. RESULTS: Caregiver facilitation of health-care was positively related both to the presence of personal and social resources and to burden levels. The results suggest that quality caregiving can coexist with burden, provided that ample caregiver resources are present. The most important resources were caregiver sense of competence and support from the professional health-care provider. DISCUSSION: Caregiving burden is not readily reducible, given the chronic nature of older people's health problems. However, caregiver resources can be bolstered, particularly by health professionals. Nurses, who are orientated to holistic family centred care, are especially well-suited for this important intervention. PMID- 12581099 TI - What areas of cancer care do Norwegian nurses experience as problems? AB - BACKGROUND: As the number of people diagnosed with cancer increases and the treatment of cancer changes and improves, the challenges and problems nurses experience in clinical practice are also changing. AIMS: To describe the problems that nurses in cancer care face in their day-to-day practice. Such information can be used to improve the quality of nursing care. DESIGN: A questionnaire was distributed to half of the members of the Norwegian Society of Nurses in Cancer Care (n = 464). Instrument. The survey questionnaire had previously been used in a Canadian study. The nurses were asked to consider 80 different areas relevant to cancer nursing and to indicate the extent to which each posed a problem for them in clinical practice. The sex, age and educational level of the nurses were also recorded, together with the work setting and number of years they had been employed in nursing. RESULTS: The response rate was 43% (199 of 464). The area the nurses experienced as being the greatest problem in their clinical practice was patients' anxiety. Problems connected with nutrition, the development of the cancer itself and grieving, and symptoms like nausea and vomiting and fatigue were also frequently rated as problems. The results also show some significant correlations between the rating of problems and variables such as work setting, years in practice and educational level. CONCLUSIONS: The present study shows that nurses in cancer care mainly experience psychological issues as problems in their clinical practice, which confirms previous research. The impact of work setting, years in clinical practice and education was surprisingly small. As the majority of nurses today will meet cancer patients during their professional careers, further research is needed in this area. PMID- 12581100 TI - Strategies for resolving aboriginal adolescent pregnancy in eastern Taiwan. AB - BACKGROUND: Adolescent pregnancy has become a main issue in the health care system for aborigines in eastern Taiwan. Using aboriginal nurses to provide information on sexual behaviour may have potential as a means of promoting healthy sexual practices among aborigines. Aim. To explore aboriginal nurses' perspectives on strategies for resolving the high prevalence rate of Aboriginal adolescent motherhood in eastern Taiwan. DESIGN AND METHODS: A qualitative research design was employed and intensive individual interviews and focus groups were conducted among a convenience sample of aboriginal nursing staff in eastern Taiwan. Content analysis was used to analyse the data. RESULTS: Nine strategies were identified. They can be divided into family, school and society aspects. The family aspect included 'enhancing parents' sex education' and 'enhancing the understanding of older people about sex education in villages'. The school aspect included 'enhancing sex education in school', 'offering interrelationship courses for adolescents', 'offering gender issue courses for adolescents' and 'enhancing school teachers' sex education training'. The social aspect included 'advocating the sense of family in church', 'advocating social norms in church' and 'discussing appropriate sexual behaviour in adolescent church fellowships'. CONCLUSIONS: Health care providers can design intervention programmes to reinforce the ability of family, school and society to deliver sex education. In addition, training and cooperating with Aboriginal nurses to execute these programmes may also decrease teenage pregnancy rates in the future. PMID- 12581101 TI - Traditional postpartum practices among Thai women. AB - BACKGROUND: Culture is often related to notions of well-being, illness, healing and health that inform individuals in their day-to-day activities. The postpartum period is noted for traditional practices related to rest, healing and the consumption of food and drinks, but a contemporary view of these practices is needed. AIMS: To gain an understanding of the traditional practices that Thai women follow in relation to postpartum care and the rationales underpinning such practices. METHODS: This descriptive study surveyed 500 Thai women living in Ubon Ratchathani, Thailand and attending their first postpartum hospital clinic appointment. A self-completion questionnaire was specially developed and pilot tested, and then administered to women attending the clinic. DATA ANALYSIS: Descriptive statistics were used in relation to the incidence of particular behaviours. Chi-square analyses were conducted to determine relationships between demographic characteristics and traditional practices. RESULTS: The majority of Thai women adhered to traditional postpartum practices related to the notion of regaining 'heat'. These included 'lying by fire', food restrictions, taking hot baths and consuming hot drinks. Other activities involved not exposing the body to heat loss by keeping covered, not shampooing the hair, avoiding the wind and sexual abstinence. Younger, less educated, primiparous women were more likely to report traditional practices. Mothers and mothers-in-law were most influential in recommending these behaviours. CONCLUSIONS: Traditional postpartum practices are still dominant in contemporary Thai culture and are perpetuated by close female family relatives. Health professionals need to be aware of clients' culture and consider the extent to which professional care complements the mothers' traditional beliefs. Nurses need to educate women about the benefits of contemporary postpartum care and to provide strategies to help them to integrate their beliefs and the practices recommended in contemporary health care practice. PMID- 12581103 TI - The Delphi technique: myths and realities. AB - AIMS: To outline the key concepts and principles of the Delphi technique. METHODS: Reference is made to a selection of studies that illustrate a variety of methodological interpretations. Drawing on Heshusius's concept of 'goodness criteria', particular emphasis is given to the question of scientific merit and means of evaluation. CONCLUSIONS: Although the technique should be used with caution, it appears to be an established method of harnessing the opinions of an often diverse group of experts on practice-related problems. PMID- 12581102 TI - Parents' use of nonpharmacological methods to alleviate children's postoperative pain at home. AB - STUDY BACKGROUND: Nonpharmacological methods are stated to be effective in alleviating children's postoperative pain when used as an adjuvant to analgesics. However, little is known about how these methods are used by parents at home. PURPOSE OF THE STUDY: The purpose of this study was to describe parents' use of nonpharmacological methods at home in 1-6-year-old children's pain alleviation after minor day surgery. METHODS: Mothers (n = 201) and fathers (n = 114) whose child had undergone day surgery in 10 Finnish hospitals between October 2000 and September 2001 filled in a questionnaire including a Visual Analogue Scale, Parents' Postoperative Pain Measure and a subscale consisting of 25 items measuring parents' use of several nonpharmacological pain alleviation methods with their children at home after day surgery. RESULTS: The most frequently used nonpharmacological pain alleviation methods were holding the child on the parent's lap, comforting the child and spending more time with them. Differences were found in mothers' and fathers' use of these methods. In addition, several methods were used more with girls than with boys. Significant relationships were found between parents' use of nonpharmacological pain alleviation methods and children's pain intensity and pain behaviour. CONCLUSIONS: Parents used several nonpharmacological pain alleviation methods with their children. However, their implementation seemed to be gender-related and limited to methods that were familiar to the parents from everyday life. The findings can be used in advising parents to use nonpharmacological pain alleviation methods at home after surgical procedures. Further research using experimental designs is needed into the effectiveness of these methods. PMID- 12581104 TI - Putting the pieces together: teaching undergraduate research from a theoretical perspective. AB - PROBLEM/PURPOSE: Baccalaureate graduates are expected to utilize research across a wide variety of practice settings. While the literature reports a variety of teaching approaches, few studies examine baccalaureate students' comprehension of research content. Teaching techniques that focus on a conceptual or theoretical approach may foster research comprehension. Therefore, the purpose of this paper is to evaluate teaching/learning outcomes of an undergraduate nursing research course designed from a conceptual or theoretical approach. PROCEDURE/FINDINGS: Two classes of senior baccalaureate nursing students (n = 47) at a private institution, whose curriculum was based on the Roy adaptation model, were surveyed in 1990 and 1991 at the end of their undergraduate research course. The survey tool consisted of seven three-point Likert scale questions, four open ended questions, and one unstructured comment. Findings showed that 72% strongly agreed that they would continue to read nursing articles in their practice field, 57% disagreed that they were intimidated by research language, and 55% agreed that they trusted their ability to use and utilize nursing research in practice. The most helpful learning activity was the research critique (34%) followed by group work (28%). The support of the teacher and Instructor's use of own research examples was also seen as most helpful (36%), while abstract cards (8%) were least helpful. Nonetheless, 23% requested more group activities, 13% wanted more class examples, and 11% asked for more time to comprehend definitions. IMPLICATIONS: Students who approached research from the perspective of a nursing conceptual framework indicated that they put the pieces of the research puzzle together by working in groups, being supported by the Instructor, and learning from a variety of teaching methods. PMID- 12581105 TI - Swedish nursing students' attitudes to and awareness of research and development within nursing. AB - BACKGROUND: Nurses' attitudes towards research have been illuminated in many studies. However, there is still a need for more knowledge about the attitudes and awareness of undergraduate nursing students. AIMS: To investigate Swedish undergraduate nursing students' attitudes towards and awareness of research and development within nursing, and to illuminate factors that may have an impact on their attitudes and awareness. METHODS: A questionnaire consisting of three parts: demographic data, attitude scale, and research awareness was used. Parametric and nonparametric statistics were used. Two hundred and one students participated in the study and the response rate was 77%. Cronbach's alpha test on the seven factors was between 0.52 and 0.80. RESULTS: Most of the students had positive attitudes towards nursing research. More than half (60%) of the students expected to make use of nursing research sometime in the future and about 25% had never read any of the scientific nursing journals. The majority (77%) of the students were acquainted with at least one Swedish researcher, but few had attended a doctoral dissertation. Students with an interest in some particular development or research area in nursing were significantly more positive and expected to make use of nursing research more frequently. Other variables that were found to have some impact upon attitudes were gender, age and additional academic studies. CONCLUSIONS: Nursing students had an overall positive attitude towards nursing research and their interest in a particular development or research area was the most important variable for their attitudes and expected use of nursing research in the future. Therefore, it would seem to be important to encourage the students' interest in specific development/research areas of nursing during their education. In spite of the nursing students' positive attitudes to nursing research, the study demonstrates the theory-practice gap is still present to some degree. PMID- 12581106 TI - Nurses who left the British NHS for private complementary medical practice: why did they leave? Would they return? AB - BACKGROUND: Although the successful retention of National Health Service (NHS) nursing staff is a policy concern, evidence suggests that substantial numbers of registered nurses continue to leave the profession, and some to own and run small caring-related business. At the same time, private complementary medicine has undergone a rapid expansion in recent years, and initial impressionistic evidence suggests that many therapists are former nurses. AIMS: The paper investigates the motivations and experiences of nurses who have either left nursing altogether, or substantially reduced their involvement, and who now practise complementary medicine in the private sector. It also investigates the issue of whether they would consider returning to practise complementary medicine within the NHS. METHODS: A combined questionnaire (n = 63) and semi-structured interview (n = 11) survey was used to collect detailed attitudinal data from respondents. The analytical approach to the interview data used a form of constant comparative analysis which was derived from the principles of grounded theory. In particular, the interviews were developed and refined from each other as they progressed. RESULTS: The majority of therapists operate independently, either from their own homes or from small rented premises. They were originally motivated by a disillusionment with the health service and particular aspects of their former jobs. However, equally they were attracted by the therapies themselves, the conceptual paradigms which underpin them and the different experiences of practice. Despite stating a disillusionment with the NHS, most respondents expressed a cautious willingness to potentially re-enter the NHS to practise complementary medicine. CONCLUSIONS: Many practising complementary therapists are registered nurses who may be willing to re-enter the formal health service. It is therefore argued that a greater synergy between orthodox and complementary medicine may be one way of addressing shortages of nursing labour, and in particular this form of drop-out, whilst meeting evolving consumer health care demands. PMID- 12581109 TI - Who owns the child in hospital? A preliminary discussion. AB - AIM: To 'own' a person is considered an infringement of human rights, but we suggest that concepts of ownership influence interactions between parents and staff when a child is admitted to hospital. This paper aims to stimulate debate and contains an explanation of the exploration of the literature for research and discussion of ownership of the child. METHOD: A wide variety of library indexes, databases and populist media were examined although it was impossible to examine all literature which may have contained references to this topic, and, apart from databases which contained abstracts in English, we could not include literature written in any language other than English, Swedish, and Icelandic. FINDINGS: We found no research that examines how concepts of ownership of a child affects communication between health professionals and parents and, ultimately, the delivery of health care. This paper begins discussion on the issues. DISCUSSION: Historical literature shows that ownership of humans has been a part of many cultures, and parents were once considered to own their children. Ownership of another has legal connotations, for instance in guardianship struggles of children during marriage breakup and in ethical debates over surrogacy and products of assisted conception. Within health care, it becomes a contentious issue in transplantation of body parts, in discourse on autonomy and informed consent, and for religious groups who refuse blood transfusions. In health care, models such as family centred care and partnership in care depend on positive communication between parents and staff. If a hospital staff member feels that he/she owns a child for whom he/she is caring, then conflict between the staff member and the parents over who has the 'best interests of the child' at heart is possible. CONCLUSION: We encourage debate about concepts of who owns the hospitalized child - the parents or the staff? Should it be argued at all? Is the whole concept of ownership of another, be it adult or child, the ethical antithesis to modern beliefs about human rights? Comment on this issue is invited. PMID- 12581110 TI - The art of developmental care in the NICU: a concept analysis. AB - INTRODUCTION: The survival rate of infants born prematurely and hospitalized in Neonatal Intensive Care Units (NICUs) has increased due to improvement of the technology in obstetrics and neona tology that allows saving preterm infants at earlier gestational ages. Preterm infants are hospitalized in a care environment which is quite different from the maternal womb. Therefore the application of developmental care in NICUs is critical to promote the development of preterm infants while they are hospitalized. AIM: This concept analysis contributes to the body of knowledge in developmental care and clarifies its understanding as well as its involvement in neonatal practice and research. METHODS: Using the method of concept analysis of Walker and Avant, this article identifies the definitions and uses of the concept of developmental care, exposes its main attributes and introduces a model case representing the acknowledged attributes. The antecedents of the concept and consequences of the application of developmental care are discussed. CONCLUSION: This concept analysis provides guidelines for nurses and health professionals to establish a 'developmental care' environment which is conducive for preterm infants. Moreover, it suggests conceptual and operational foundations of developmental care for clinicians and researchers in the neonatal field. PMID- 12581111 TI - Predictors of maternal competence by first-time mothers when the child is 8 months old. AB - BACKGROUND: Maternal role attainment is largely a cognitive and social process that is influenced by the cultural and family context and the mother's and child's traits and characteristics. Feedback from a social network is important to validate competent performance. AIM: To ascertain factors that contributed to the maternal competence of first-time mothers when the child was 8 months old. METHODS: Data collection was by structured questionnaires. The sample consisted of 248 first-time mothers who completed the questionnaires when their child was 8 months old. The data were analysed using stepwise regression analysis. RESULTS: According to the multivariate analysis, nine predictors remained in the model to explain maternal competence. The more balanced the mother's state of mind, the better she felt about succeeding in child care, and the more easy going the child was, the greater was the mother's competence. The more the mother felt isolation from society, the less was her maternal competence. Younger and healthier mothers, and those who readily accepted their child showed better competence. Also, if the mother still breast-fed her child, the more concrete support she received from the support network, the better was her maternal competence. CONCLUSION: The findings suggest important factors that contribute to the maternal competence of first-time mothers include mothers' resources, ease of caring for the child, breast-feeding and social support received from those close to them. PMID- 12581112 TI - Health beliefs, perceived self-efficacy, and breast self-examination among Thai migrants in Brisbane. AB - BACKGROUND: Women in Thailand have a relatively low risk of developing breast cancer; however, death rates from breast cancer are increasing. Rates in many migrant groups are also known to be on the increase. Little is known about breast cancer screening, particularly breast self-examination (BSE), among Thai migrant women in other countries. In Australia, non-English-speaking-background migrants are known to be low users of preventive health services. AIMS: To investigate, using the health belief model (HBM) and self-efficacy as a theoretical framework, the use of BSE in a recent migrant group, Thai women in Australia, and to identify sociodemographic variables that influence the women's regular use of BSE. METHODS: In 1998, a cross-sectional study was conducted among 145 Thai women in Brisbane recruited through a snowball-sampling method, which used personal contacts and key persons within the Thai community. The study was approved by the University Human Ethics Review Committee. Data were collected through designed closed-ended questions. RESULTS: Only 25% of the women performed BSE regularly. HBM indices were strongly associated with BSE. Beliefs in high personal susceptibility to breast cancer strongly increased the likelihood of BSE. After adjusting for potential confounding factors, cues or triggers to undertake BSE and self-efficacy, or the ability to do BSE were found to be important determinants of regular BSE. Study limitations, including data collection methods, are discussed. CONCLUSION: A low percentage of women practised BSE regularly. The HBM is a useful framework for identifying factors influencing the use of BSE. Strategies that increase the confidence of women to undertake preventive health behaviour or increase self-efficacy are likely to increase their regular screening for breast cancer. PMID- 12581113 TI - Cancer patients' decision-making regarding treatment and nursing care. AB - BACKGROUND: Patient participation in decision-making has produced many debates among health care professionals. The research evidence concerning patient participation in decision-making is not clear, and shows conflicting results. AIMS: To identify to what extent cancer patients participate in decision-making and to what extent background characteristics, information obtained and relationships with staff explain cancer patients' participation in decision making. DESIGN/METHOD: A structured questionnaire based on earlier research and qualitative interviews was designed. The questionnaire was completed by 273 cancer patients who were inpatients or outpatients in haematology, oncology clinics and other clinics at two university hospitals in Finland. The questionnaire consisted of five areas: (1) demographic data; (2) mood; (3) information obtained; (4) relationships with staff; and (5) decision-making. The scale used was ordinal. The data were analysed using descriptive statistics and nonparametric tests such as Mann-Whitney U-test, Kruskall-Wallis, and Wilcoxon matched-pairs signed-rank tests. RESULTS: The majority of the respondents perceived that they had participated in making treatment decisions at least to some extent, together with the physician. In decision-making about nursing care, respondents felt that they had participated most in decisions about personal hygiene, rest and sleep. Respondents' physical condition, marital status, age and time since diagnosis, as well as information obtained and relationships with staff, were associated with participation in decision-making. CONCLUSION: This study provided evidence that some cancer patients participated in decision-making and felt participation to be important. Staff played a crucial role in patient participation in decision-making. In future, staff should work to improve cancer patients' opportunities to participate in decision-making in order that those who wish to be involved can do so. PMID- 12581114 TI - Acute pain teams and the management of postoperative pain: a systematic review and meta-analysis. AB - BACKGROUND: The introduction of acute pain teams (APTs) in every hospital performing surgery in the UK has been recommended in order to reduce postoperative pain. However, recent evidence suggests that many APTs are under resourced. Purchasers may be more prepared to invest in these services if they are persuaded that they result in measurable improvements in patient outcomes. AIM: A systematic review of the literature and meta-analysis were performed to determine the effectiveness of APTs in improving the quality of analgesia and other postoperative outcomes of adult patients undergoing surgery. METHODS: A broad search strategy using the terms 'pain team' and 'pain service' was adapted for a variety of databases. Key journals were hand-searched and reference lists of selected reports were reviewed. Subject experts and study authors were contacted. Studies describing the impact of the APT/acute pain service (APS) on postoperative pain relief, other postoperative outcomes or the processes of postoperative pain were included. Study quality was assessed using a multidimensional instrument. A broad qualitative overview of the included studies was conducted. Continuous outcome data for pain in the first 24 hours postoperatively (in one case worst pain at 24-48 hours) were pooled. RESULTS: Fifteen studies were included in the review. There were considerable differences in study design and quality, the nature of the APT and the outcomes measured. Of the nine studies measuring pain, it was possible to present data as Standardized Mean Differences for only four studies. Quantitative synthesis indicates a statistically significant overall estimate of effect using a fixed effects model only. LIMITATIONS: Only published studies in English were included. Study inclusion decisions and data extraction were performed by one reviewer only. CONCLUSION: There is insufficient robust research to assess the impact of APTs on postoperative outcomes of adult patients or on the processes of postoperative pain relief. PMID- 12581115 TI - Patient injury and physical restraint devices: a systematic review. AB - OBJECTIVE: To investigate physical restraint-related injuries. Areas of interest were the prevalence of injury, types of injuries, risk of sustaining an injury and specific restraint devices associated with injury. DEFINITIONS: Injury in the context of this review was considered to be either direct injury, such as lacerations and strangulation, or indirect injury considered to be an adverse outcome such as increased mortality rates or duration of hospitalization. METHOD: A comprehensive search was undertaken that involved all major databases and the reference list of all relevant papers. To be included in the review studies had to involve people in acute or residential care settings and report data related to injury caused by restraint devices. A number of different types of research designs were included in the review. The findings of studies were pooled using odds ratio and narrative discussion. RESULTS: The search identified 11 papers reporting the findings of 12 observational studies. These studies were supplemented with the findings of a number of other types of studies that reported restraint-related data. The review highlights the potential danger of using physical restraint in acute and residential health care facilities. Observational studies suggest that physical restraint may increase the risk of death, falls, serious injury and increased duration of hospitalization. However, there is little information to enable the magnitude of the problem to be determined. DISCUSSIONS: Many of the findings highlight the urgent need for further investigation into the use of physical restraint in health care facilities. Further research should investigate the magnitude of the problem and specific restraint devices associated with injury. However, given the limited nature of the evidence, this association should be investigated further using rigorous research methods. PMID- 12581116 TI - Portfolios and assessment of competence: a review of the literature. AB - BACKGROUND: The literature review presented here was conducted as part of an English National Board for Nursing, Midwifery and Health Visiting funded project to evaluate the use of portfolios in the assessment of learning and competence. Initial consideration of the topic revealed the need to clarify the terminology and approaches used to assess competence using portfolios, and therefore the literature review was conducted to inform the study. AIMS: To clarify definitions, theoretical bases and approaches to competence and the use of portfolios in the assessment of learning and competence in nursing education. METHODS: A comprehensive literature review was conducted using the CINAHL and MEDLINE databases and the keywords competenc*, portfolios and nursing. Articles published in the period 1989-2001 in English were obtained and their reference lists scrutinized to identify additional references. Twenty articles were found using a combination of the keywords competenc* and portfolios, and 52 using the combination portfolios and nurse education. Articles were included in the review if they focused on the use of portfolios in nursing, and those concerned with professional or transitional portfolios were excluded. This article will analyse definitions of and approaches to competence and its measurement and to portfolios and their use as discussed in the articles identified. RESULTS: Three approaches to competence were identified, each with its appropriate forms of assessment. With regard to portfolios, a number of definitions were again found, but there was a consensus that the theoretical basis of their use is theories of adult learning. A number of reasons for and advantages and disadvantages of their use were found, as well as varying ideas about what a portfolios should consist of and how it should be assessed. CONCLUSION: A holistic approach to competence seems to be compatible with the use of portfolios to assess competence in nursing students, but the concept and its implementation is still evolving. A variety of assessment methods are needed for assessment and portfolios appear to have the potential to integrate these. Reflection is an essential component of a portfolio, as are the student-teacher relationship and explicit guidelines for constructing the portfolio. Issues of rigour in assessment of portfolios need to be addressed, but the assessor's professional judgement will inevitably enter into this assessment. PMID- 12581117 TI - Supporting carers of people diagnosed with schizophrenia: evaluating change in nursing practice following training. AB - BACKGROUND: United Kingdom legislation and clinical standards for schizophrenia challenge nurses to re-examine the support that they provide to carers. Nurses are in a key position to provide this support but may lack the necessary skills to do so. The training programme evaluated in the present study aimed to address this problem. STUDY AIM: To evaluate change in clinical practice brought about by post-registration training for mental health nurses in supporting carers of people diagnosed with schizophrenia. DESIGN/METHODS: The study was undertaken in collaboration between the Universities of Dundee and Glasgow, and Tayside National Health Service (NHS) Trust (Scotland). Respondents were nine nurses who completed training and then delivered a planned programme of support to carers. Data on nursing practice were gathered through semi-structured interviews with nurses before training and after providing support. Following the support intervention, carers also commented on the nurses' practice. FINDINGS: Eight of the nine nurses reported changes in practice in five key areas: They built collaborative relationships with carers, developed a carer focused approach to their practice, acknowledged and supported the carer role, and made progress in identifying carer needs and accessing resources to meet these needs. Nurses experienced difficulties supporting carers who had mental health problems or previous negative experiences of services. Those who lacked community experience also found it difficult to adjust to working in a community setting. Although clinical supervision helped them to work through these difficulties, they remain largely unresolved. CONCLUSIONS: Findings from this study indicate that appropriate training may enable nurses to improve the support provided to carers of people diagnosed with schizophrenia. This study represents an important stage in determining the nature of support offered to carers by nurses. While developed to help nurses to meet clinical standards set for schizophrenia in the UK, findings may have clinical significance for nurses in other countries. PMID- 12581118 TI - In the field: notes on observation in qualitative research. AB - BACKGROUND: Observation is used in research in two ways - structured and unstructured. Which of these methods to choose depends on the research question but will be defined predominantly by the paradigm underlying each study. Positivistic research generally uses structured observation and interpretist/naturalistic paradigms use unstructured observation. This paper is concerned with unstructured observation. AIM: To discuss the importance of unstructured observation as a research method and to examine critically the problems associated with both access and field notes. APPLICATION TO NURSING: Although unstructured observation has been used in nursing research, it is less favoured than the interview. Furthermore, although some of the dilemmas concerned with observation such as the role of the researcher, covert research and informed consent have received attention in the nursing literature, other aspects remain relatively unexplored. In particular little has been written concerning access to the field and field notes. Both of these aspects have practical and theoretical considerations that require the attention of the researcher. This omission may partly explain nurses' apparent reluctance to use unstructured observation in their studies. This paper seeks to redress this situation. PMID- 12581119 TI - Testing the law: Swiss nurses go to court to fight for equal pay. PMID- 12581120 TI - Building on the work of our colleagues. PMID- 12581121 TI - Asking nurses to share their authentic experiences: a conversation with Dr Jane Robinson [interviewed by Jan Harrington]. PMID- 12581122 TI - Poverty - what can nurses do? PMID- 12581123 TI - Reflections on the Judgement Axis, ICNP beta. International Classification of Nursing Practice. AB - This article presents questions, reflections and proposed resolutions regarding the Judgement Axis of the International Classification of Nursing Practice, Beta Version (ICNP beta). The Judgement Axis, in its present format, consists of an alphabetical listing of terms that are meant to reflect the clinical opinion of a nurse relating to the state of a nursing phenomenon appearing in Axis A of the ICNP beta. A review of literature indicates that judgement can be more extensive. Not considered in the development of the Axis is the fact that judgement can be clinical and actuarial. Actuarial judgement is predictive and based on statistics. Clinical judgement is patient-centred, creative and intuitive. Both are essential in defining nursing judgement. The list presents 49 modifiers in nominal form. Each of these is subordinated, in alphanumeric descending order, by qualifiers that do not conform to the concept of lexical hierarchy. We propose that the 49 terms which appear in the present listing as unique ordinates should be clustered into major categories with synonyms listed as subordinates of the major cluster headings. The qualifiers of degree, which are presently identified as subordinates in Axis B, might be presented in matrix format, providing a scale of degree of the modifier for the nurse. PMID- 12581124 TI - The role of nurses in the human immunodeficiency virus/acquired immune deficiency syndrome policy process in Botswana. AB - In Botswana, there is dearth of literature on the role of nursing in health-care policy and resource allocation and yet nurses constitute the majority (85%) of health manpower. The health-care delivery system depends mostly on nurses for service provision. There were two main purposes of this study: first, to gather descriptive data from major key players (with particular emphasis on nurses) concerning knowledge of the policy process and resource allocation for management and care of clients with human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) in Botswana; and, second, to identify nurse characteristics (e.g. position, education, experience, job category) associated with motivation to influence health-care policy in HIV/AIDS management and care in Botswana. A policy process conceptual framework was used to guide data collection and analysis. A case-study research method was used to conduct in depth interviews from a purposive sample of 19 policy makers, and a survey questionnaire was used to collect data from a purposive sample of 95 registered nurses from six study sites in Botswana. The study findings indicate minimal participation of nurses in health-care policy process and resource allocation. The demographic variable of position was a predictor of the involvement of nurses in policy and in budgetary decisions. Both survey and interview data indicated that this minimal participation of nurses in the policy process resulted in implementation problems, thus compromising service provision. Implications of the findings for the nursing profession, nursing practice and policy, which address the importance of nurses' involvement, are discussed. PMID- 12581125 TI - Culture-derived strategies of a paediatric home-care nursing specialty team. AB - This is an ethnographic study of one paediatric home-care nursing specialty team that cares for children and their families affected by perinatal human immunodeficiency virus (HIV) disease. Observations made by the investigator as part of a larger study suggested that the intricacy and breadth of nursing knowledge and actions that were actually used far exceeded what the standard documentation recorded. Spradley's Developmental Research Sequence, combining methods of participant observation and interviewing, was used to answer the question: What nursing strategies are used in managing the health needs of HIV exposed infants in a home-care programme? Thematic analyses revealed that the visit process had four essential elements, as perceived and lived by the nurses: the pursuit; the connection; the work; and the disengagement. This full scope of nursing strategies for assisting and empowering these families remains largely invisible, limited to verbal transmission to family caregivers and among nurses. It needs to be documented, claimed and further studied by nursing. PMID- 12581126 TI - Self-rated performance of quality specialists in Korea. AB - This study was conducted to explore the self-rated performance level of quality specialists in Korea in order to provide information to direct the future needs for education and role orientation. The formal identification and appointment of quality specialists in health-care organizations in Korea began in the mid-1990s. The majority of quality specialists, appointed by their hospital, assumed positions without undergoing the prerequisite training or education. Extensive variation in performance has been recognized. The sample (n = 29) in this study included all quality specialists at hospitals in Seoul and Kyung-Gi province. Their performance was rated using a postal questionnaire consisting of a set of activities in five domains (policy development, data collection and analysis, communication and report, education and support, and administration). The response rate was 100%. The respondents had an average of 25.5 months of experience as quality specialists. Self-rated performance of quality-related activities demonstrated wide variation, independent of personal, departmental, or hospital-specific characteristics. The respondents rated their performance highly in the area of communication and reporting. The performance of administrative activities was rated lowest. Perceived differences between desired and actual performance were statistically significant in all activities of each domain, and were largest in policy development, and next in education and support. Self-rated performance among the five domains demonstrated positive, significant correlations. Hence, the findings imply the necessity of a formal education for quality specialists to accelerate balanced growth in all domains of quality activities. PMID- 12581127 TI - Culture, values and racism: application to nursing. AB - The prevention and management of cancer have been identified as health priorities in most countries. In order to achieve this, many oncology nurses need to face the additional challenge of caring for patients from different cultures. This article progresses beyond previous literature, which generally adopts a solely multicultural approach, and explores the concepts of culture, values and racism with inter-related concepts such as self-awareness, cultural encounter, skills and cultural knowledge and their relationship to cancer nursing practice. The case is then argued for enhancing patient care through the application of these concepts to nursing practice. PMID- 12581128 TI - Endometrial Cancer. PMID- 12581129 TI - Breast Cancer. PMID- 12581131 TI - Miscellaneous. PMID- 12581130 TI - Genetic Testing. PMID- 12581132 TI - Introduction. PMID- 12581134 TI - Death receptors and their role in dermatology, with particular focus on tumor necrosis factor-related apoptosis-inducing ligand receptors. PMID- 12581135 TI - Kaposi's sarcoma in renal transplant patients: experience at the Cruces Hospital in Bilbao. AB - BACKGROUND: Kaposi's sarcoma (KS) in renal transplant recipients (RTRs) probably arises from a complex interplay of multiple factors. OBJECTIVE: In order to analyze the prevalence of KS in patients transplanted at the Cruces Hospital in Bilbao, together with their clinical features, treatment, and etiologic factors, we performed a study using the registry of RTRs in our center. METHODS: The records of 1,230 kidney transplant patients at the Cruces Hospital between 1979 and 1998 were reviewed. Immunosuppressive therapy was reduced once a diagnosis of KS was made. A nested polymerase chain reaction was used to detect human herpesvirus 8 (HHV-8) DNA in the biopsy tissue. The DNA was extracted from fresh tissue (n = 2) or from formalin-fixed, paraffin-embedded specimens (n = 5). RESULTS: Six cases of KS were diagnosed. All patients with cutaneous KS improved with a reduction in immunosuppressive drugs. HHV-8 was detected in 100% (2/2) of the frozen biopsies and 20% (1/5) of the formalin-fixed samples investigated. CONCLUSIONS: Our experience indicates that a continuous state of immunodeficiency is important for the development of KS in RTRs. The association, previously described between HHV-8 and transplant-associated KS, also exists in the studied population. PMID- 12581136 TI - Antimicrobial resistance of Staphylococcus aureus isolated from impetigo patients between 1994 and 2000. AB - BACKGROUND: The incidence of strains of Staphylococcus aureus resistant to the antimicrobial agents used in treating impetigo has been increasing. AIM: To determine the antimicrobial susceptibility of S. aureus in impetigo. METHODS: We measured the antimicrobial susceptibility of Staphylococcus aureus isolated from impetigo patients between 1994 and 2000. RESULTS: The MIC50 of gentamicin was always higher than that of other antimicrobial agents until 1999. In isolates obtained since 1996, the MIC90 of gentamicin was over 12.5 micro g/mL, which is markedly higher than that found for other skin infections (folliculitis, furuncles, paronychia, phlegmone, secondary infection of eczema, dermatitis, ulcer and decubitus). There were no strains of S. aureus resistant to vancomycin and fusidic acid. After 2000, we could find only one strain resistant to minocycline and ofloxacin. CONCLUSION: Clindamycin has shown excellent activity against most S. aureus isolates between 1994 and 2000. The incidence of methicillin-resistant S. aureus was always below 20%. PMID- 12581137 TI - Febrile ulceronecrotic pityriasis lichenoides et varioliformis acuta with atypical cells. PMID- 12581138 TI - Stop female genital mutilation. A perspective: religious rites vs. cultural norms. PMID- 12581140 TI - Scleromyxedema revisited. AB - Scleromyxedema is a rare disease characterized by papular mucinous deposits, dermal fibroblast proliferation, and a monoclonal paraproteinemia. Its chronic course tends to be complicated by gastrointestinal, muscular, pulmonary, and neurologic disorders. We review the literature to identify the common systemic manifestations and suggest a clinical approach to this disorder. This should include: identification of the characteristic lesions, a skin biopsy, and serum protein electrophoresis. A thyroid evaluation may help differentiate this condition from others. An early neurologic, gastrointestinal, and cardiovascular evaluation should be considered. PMID- 12581141 TI - Large cell acanthoma. AB - The large cell acanthoma presents as a slightly scaly tan macule on photodamaged skin. Clinically, it may be difficult to differentiate from a lentigo senilis, pigmented actinic keratosis, or a flat and pigmented seborrheic keratosis. We have studied 19 cases of large cell acanthoma. Large cell acanthomas were identified histologically as having epidermal keratinocytes with nuclei roughly twice the size of adjacent epidermal or adnexal keratinocytes, and as having minimal nuclear pleomorphism. Histologic findings were compared with actinic keratosis and lentigo senilis. Melanocyte density and cellular proliferation were compared using HMB-45 staining of melanocytes and proliferating cell nuclear antigen (PCNA) staining of epidermal keratinocytes. Lentigo senilis and large cell acanthoma both showed increased numbers of melanocytes, as identified by HMB 45 staining. Actinic keratosis shows a statistically increased proliferation rate, as identified by PCNA staining. On the basis of clinical, histologic, and immunohistochemical staining similarities, we believe that large cell acanthoma should be considered as a reaction pattern, possibly related to lentigo senilis. PMID- 12581142 TI - Molecular insights into the hair follicle and its pathology: a review of recent developments. AB - In the last few years by means of the elucidation of the human genome and the acquisition of powerful investigative tools we have begun to understand the molecular basis of hair follicle growth control. In this article I will describe some of the salient recent contributions to the field and review the implications these findings have had on our understanding of mechanisms in dermatology and dermatopathology. PMID- 12581143 TI - Felix Pinkus: dermatologist, dermatopathologist, and artist. PMID- 12581144 TI - Assessment of histologic criteria in the diagnosis of mycosis fungoides. AB - BACKGROUND: The histologic diagnosis of early mycosis fungoides (MF) can be difficult to establish in many instances because the subtle changes observed in patches of MF are also present in many inflammatory dermatoses. METHODS: To assess the frequency and significance of many of these histologic parameters, we retrospectively reviewed 50 slides from patients with documented MF in patch, plaque, and tumor stages. The diagnosis of MF was unequivocally established either by the progression of patients to advanced stages of the disease or by indubitable histologic findings. In the second phase of the study, we compared the histologic parameters observed in 24 patch stage MF patients with those in 24 non-MF patients. The non-MF group were patients whose pathologic pattern was suspicious for MF, but who definitely did not have MF on clinical grounds. The two groups were matched by histologic pattern. Two different observers evaluated the slides and the intensities of 32 histologic parameters were graded on a four point scale to minimize the subjective variability in the histologic reports. RESULTS: On univariate analysis, the following parameters achieved significance in distinguishing MF from non-MF: Pautrier's microabscesses, haloed lymphocytes, disproportionate epidermotropism, epidermal lymphocytes larger than dermal lymphocytes, hyperconvoluted lymphocytes in the epidermis and dermis, absence of dyskeratosis, and papillary dermal fibrosis. None of these features proved to have additional discriminating power on multivariate analysis. CONCLUSIONS: The efficacy of single histologic features in the diagnosis of early MF is generally poor and, to discriminate MF from its inflammatory simulators, a combination of cytologic and architectural features must be used. PMID- 12581145 TI - Treatment of mycosis fungoides/Sezary syndrome: the University of California, San Francisco (UCSF) approach. PMID- 12581146 TI - Value of histopathology in vitiligo. AB - BACKGROUND: Histopathologic studies of vitiligo have been rather limited in number, thus the microscopic features of this common disorder are not very well known. SUBJECTS AND METHODS: Skin specimens from 20 human vitiligo patients and skin specimens from five equine vitiligo patients were studied by light and transmission electron microscopy. RESULTS AND DISCUSSION: Absence of melanocytes, increased number of Langerhans' cells, epidermal vacuolization, thickening of the basement membrane, T-cell inflammatory infiltrate, and neural alterations were noted in the vitiligo lesions. These results may explain the development of depigmentation, but do not yet fully explain why and how this occurs. PMID- 12581147 TI - Wells' syndrome: a clinical and histopathologic review of seven cases. AB - Wells' syndrome, or eosinophilic cellulitis, is characterized clinically by an acute dermatitis resembling cellulitis, which evolves into violaceous plaques that resolve spontaneously without scarring. The histopathologic features are dynamic, starting with dermal edema and infiltration of eosinophils, the development of "flame figures," and finishing with the appearance of phagocytic histiocytes. We present the clinical and histopathologic features of seven cases of eosinophilic cellulitis. PMID- 12581148 TI - Comparative evaluation of different hair removal lasers in skin types IV, V, and VI. AB - BACKGROUND: Lasers permit treatment of unwanted excess hair with less discomfort than other methods of epilation. Many lasers with different parameters are now available from which the dermatologist can choose. Improved clinical results are made possible by the high specificity and selectivity of the laser systems to pigmented hair because of the use of an appropriate wavelength with the proper pulse and duration. OBJECTIVES: We aimed to compare the results of treatment of skin types IV, V, and VI using three different laser systems. METHODS: One hundred female patients were compared using different laser systems: 35 patients underwent epilation using a Nd-Yag laser, 33 patients using an Alexandrite laser, and 32 patients using a Diode laser. RESULTS: Follow up 12 months after the multiple treatments (three to six sessions) showed an insignificant difference between these three groups (35-40%). CONCLUSION: Our findings indicate that all three laser systems tested can be used for dark skin; however, one should select a system that minimizes side-effects, primarily hypo- and hyper-pigmentation, especially when used for skin types IV, V, and VI. PMID- 12581149 TI - Apocrine carcinoma of the vulva in a band-like arrangement with inflammatory and telangiectatic metastasis via local lymphatic channels. AB - BACKGROUND: Primary adenocarcinomas of the vulva have been classified as sweat gland carcinomas, extramammary Paget's disease, and primary breast carcinomas of the vulva. They share some common histopathologic features. METHODS: We describe a 72-year-old Japanese woman with apocrine carcinoma of the vulva and local lymphatic metastasis. RESULTS: The patient presented with a bruise on her inguinal area. Physical examination revealed a 4 cm x 7 cm, dark-red, irregularly elevated tumor on the left labium majora. Dome-shaped, flesh-colored, small papulovesicles were scattered on the abdomen, accompanied by erythema and induration. The lesion showed a band-like arrangement. General examination revealed multiple bone metastases, particularly in the spine. Microscopic examination revealed a moderately differentiated adenocarcinoma with signet ring cells. A few pagetoid clear cells were present in the hypertrophic epidermis. The peripheral papulovesicles demonstrated the same histopathologic view as in inflammatory and telangiectatic, metastatic breast carcinoma. Tumor cells were positive for various ductal and glandular markers. Estrogen and progesterone receptors were not expressed. Ultrastructural findings suggested differentiation towards apocrine or mammary glands because of the presence of an apocrine process and electron-dense mucous granules. The patient died in spite of combination chemotherapy and irradiation therapy. CONCLUSIONS: We report a rare case of apocrine carcinoma of the vulva in a band-like arrangement with local lymphatic metastasis which showed the clinical and histopathologic characteristics of inflammatory and telangiectatic carcinoma. PMID- 12581150 TI - Familial anetoderma. AB - A 31-year-old Caucasian male presented with a history of erythematous, saccular outpouchings of the skin on his back, shoulders, and upper extremities (Fig. 1). The patient reported that his mother and aunt had a similar skin disorder, which initially began with inflammation, but healed leaving lax skin in its wake. He did not recall the name of the skin condition. Physical examination revealed large confluent zones of sac-like protrusions of erythematous skin on the back and shoulders. Histopathologic examination of the excisional biopsy revealed a relatively unremarkable epidermis. Perivascular lymphocytes were present in small numbers in the papillary dermis. Adnexal structures and deep dermis remained intact. The acid-orcein-Giemsa stain highlighted the absence of elastic fibers within the mid and lower reticular dermis (Fig. 2a,b), consistent with anetoderma. PMID- 12581151 TI - A call to action for early melanoma detection. PMID- 12581152 TI - Isolation and expression of a novel mitochondrial septin that interacts with CRMP/CRAM in the developing neurones. AB - BACKGROUND: Collapsin response mediator proteins (CRMPs) and CRAM belong to the unc-33 gene family which is implicated in axon guidance and outgrowth during neural development. However, their exact roles remain largely unknown. To understand the molecular basis of CRMP/CRAM function, we have undertaken to identify CRMP/CRAM interacting proteins. RESULTS: We have identified a novel mitochondrial septin (M-septin) as one of the CRMP/CRAM interacting proteins from the developing rat brain. M-septin is a major, alternatively spliced variant of the H5 gene in developing mouse brain and its expression is up-regulated during the neuronal differentiation of embryonal carcinoma P19 cells. In COS-7 cells, M septin is specifically localized to mitochondria whereas H5 is diffusely distributed to the perinuclear cytoplasm and plasma membranes. In contrast to H5, M-septin induces the mitochondrial translocation of CRAM but not CRMP2. Finally, M-septin is found to be transiently translocated to mitochondria before the induction of the neurites and then dissociates from the mitochondria after neurite extension in P19 cells. CONCLUSIONS: Our results suggest that M-septin has a role which is distinct from H5, and together with CRMP/CRAM, may play an important role in the neuronal differentiation and axon guidance through the control of mitochondrial function. PMID- 12581153 TI - Suppression of NF-kappaB-dependent gene expression by a hexamethylene bisacetamide-inducible protein HEXIM1 in human vascular smooth muscle cells. AB - BACKGROUND: Neointima formation is a characteristic feature of atherosclerosis and post-angioplasty restenosis, in which various soluble factors and mechanical injury stimulate signalling pathways in vascular smooth muscle cells (VSMC), promoting their migration and proliferation, and the eventual formation of the neointima. The transcription factor NF-kappaB has been shown to play a pivotal role in this process. Hexamethylene bisacetamide, an inhibitor of VSMC proliferation, induces the mRNA expression of HEXIM1 (hexamethylene bisacetamide inducible protein 1). However, the protein expression and function of HEXIM1 remain unknown. RESULTS: In the present study, we demonstrated that HEXIM1 localizes in the cytoplasm and nucleus, and its nuclear expression is restricted to discrete speckled areas. Treatment of VSMC with hexamethylene bisacetamide up regulated HEXIM1 expression, not only in mRNA but also protein levels. Moreover, HEXIM1 is shown to suppress the transcriptional activity of NF-kappaB via its C terminal leucine-rich domain. A glutathione-S-transferase pull down assay indicated that HEXIM1 interacts with the p65 subunit of NF-kappaB. In VSMC, treatment with hexamethylene bisacetamide resulted in a down-modulation of the transcription of NF-kappaB target genes. CONCLUSION: We may therefore conclude that HEXIM1 plays an inhibitory role in NF-kappaB-dependent gene expression in VSMC and is the candidate of a novel therapeutic target for inhibition of VSMC proliferation. PMID- 12581154 TI - Simultaneous acceleration of the cell cycle and suppression of apoptosis by splice variant delta-6 of the candidate tumour suppressor LUCA-15/RBM5. AB - BACKGROUND: The short arm of chromosome 3 is thought to include one or more tumour suppressor genes (TSGs), since carcinoma of various tissues display deletions in this region. Many genes mapping to this region have recently been identified, including the LUCA-15/RBM5 gene. RESULTS: In this study we report the cloning from human bone marrow library of a splice variant of LUCA-15 which lacks exon 6, resulting in a frameshift and producing a truncated protein of 150 amino acids instead of 815 amino acids. This variant is widely expressed at a low level in normal tissues and is expressed at increased levels in T-leukaemic cell lines. Over-expression of this splice variant after electroporation both shortened the cell cycle and inhibited CD95-mediated apoptosis in CEM-C7 T-cells. In marked contrast, over-expression of the full length LUCA-15/RBM5 suppressed cell proliferation both by inducing apoptosis and by extending the G1 phase of the cell cycle. CONCLUSION: These results, taken together with previous observations from ourselves and others, suggest that LUCA-15 is involved in the control of both apoptosis and the cell cycle. Since oncogenesis often relies on separate changes in molecules regulating apoptosis on the one hand, and proliferation, on the other, the discovery of a candidate tumour suppressor gene which affects both processes simultaneously is likely to be of major significance. PMID- 12581156 TI - Mitogen-activated protein kinase p38 defines the common senescence-signalling pathway. AB - BACKGROUND: Cellular senescence is a state of irreversible growth arrest shown by normal cells, and has been most extensively studied in replicative senescence caused by telomere shortening. Several conditions, including oncogenic Ras over expression and inappropriate culture conditions, also induce senescence without telomere shortening. However, it remains unclear how a common set of senescence phenotypes is indistinguishably induced in various types of senescence. RESULTS: We demonstrate that p38 mitogen-activated protein kinase (MAPK) plays important causative roles in senescent cells following telomere shortening, Ras-Raf activation, oxidative stress or inappropriate culture conditions. By monitoring the kinetics of p38 activation, we suggest that p38 is activated not directly by the initial stimuli, but in response to unidentified cellular conditions caused by these stimuli. Importantly, this p38-activating condition appears to be defined quantitatively as a sum of continuous and low-level stresses, and remains even after the initial stimuli are withdrawn, which may explain the well-known irreversible nature of cellular senescence. We also show that papilloma virus E7 abolishes the p38-induced growth arrest but not other senescence-associated phenotypes, indicating the differential role of pRb in the downstream of p38. CONCLUSION: These results indicate that p38 comprises the senescence-executing pathway in response to diverse stimuli. PMID- 12581155 TI - Synaptic localization of SAPAP1, a synaptic membrane-associated protein. AB - BACKGROUND: SAPAP1 was originally identified as a protein interacting with the guanylate kinase domain of PSD-95. SAPAP1 also interacts with various proteins, including neurofilaments, synaptic scaffolding molecule (S-SCAM), nArgBP2, dynein light chain and Shank through different regions. RESULTS: We expressed various regions of SAPAP1 in hippocampal neurones. The synaptic targeting of SAPAP1 was mediated by the N-terminal region and did not depend on the interaction with PSD 95 or S-SCAM. SAPAP1 was not involved in the synaptic localization of PSD-95 or S SCAM, but affected that of Shank. The synaptic targeting of SAPAP1 was not suppressed by blocking NMDA or AMPA receptors. Fluorescent recovery after a photobleaching study revealed that SAPAP1 was immobile at synapses. CONCLUSION: SAPAP1 is a component of the static core of PSD, and its dynamics are different from those of the other PSD components, PSD-95, S-SCAM and BEGAIN. PMID- 12581157 TI - CDK- and Cdc45-dependent priming of the MCM complex on chromatin during S-phase in Xenopus egg extracts: possible activation of MCM helicase by association with Cdc45. AB - BACKGROUND: MCM and Cdc45 are required for the initiation and elongation stages of eukaryotic DNA replication. Recent studies show that a purified Mcm4/6/7 complex has DNA helicase activity. However, the biochemical function of the MCM complex and Cdc45 bound to chromatin has not been elucidated. RESULTS: We have examined the biochemical properties of MCM proteins bound to chromatin fractions using Xenopus egg extracts. Immunoprecipitation of MCM proteins extracted under denaturing conditions reveals that all six subunits of MCM and Cdc45 form a tight complex following the initiation of DNA replication, and that both CDK activity and Cdc45 are essential for the complex formation. Chromatin immunoprecipitation of MCM proteins and Cdc45 shows that a complex containing MCM and Cdc45 has a DNA helicase activity which is dependent on CDK activity and Cdc45 in the extracts. Furthermore, both the complex and the helicase activity are resistant to treatment with phosphatase and high salt. CONCLUSIONS: Following the initiation of DNA replication, a tight MCM-Cdc45 complex is formed on chromatin and its formation is closely correlated with the DNA helicase activity of chromatin immunoprecipitates containing MCM and Cdc45. We propose that the tight MCM-Cdc45 complex functions as a replicative DNA helicase in vivo. PMID- 12581158 TI - Overproduction of a conserved domain of fission yeast and mammalian translation initiation factor eIF4G causes aberrant cell morphology and results in disruption of the localization of F-actin and the organization of microtubules. AB - BACKGROUND: The recruitment of mRNA for translation involves the assembly at the 5'cap of a complex of three initiation factors: the cap binding protein eIF4E, the ATP-dependent RNA helicase eIF4A and the scaffold protein eIF4G. eIF4G mediates the binding of this mRNA-protein complex to the 43S ribosomal preinitiation complex. There is growing recognition that the components of the translational apparatus interact functionally with cytoskeletal components. Here we report specific effects of the over-expression of human and fission yeast eIF4G domains on cell morphology in Schizosaccharomyces pombe. RESULTS: A single gene encoding fission yeast eIF4G was identified and demonstrated to be essential. We have over-expressed fragments corresponding to the conserved functional domains of eIF4G. At expression levels that did not disrupt rates of overall translation or protein accumulation, a fragment of S. pombe eIF4G, 4G NOB, corresponding to the minimal region of human eIF4G required to support cap independent mRNA recruitment, was found to impair cell proliferation in fission yeast. This resulted from defects in cytokinesis, and was associated with the disruption of both microtubules and actin microfilaments. The over-expressed fragment was itself localized to the cell ends, the nuclear periphery and the septum. CONCLUSIONS: This is the first demonstration of a link between a translation initiation factor and mechanisms controlling cell morphology. The data suggest a direct or indirect interaction between the functional domains of eIF4G and cellular structures involved in cytokinesis. PMID- 12581159 TI - Instability of sensory histidine kinase mRNAs in Escherichia coli. AB - BACKGROUND: Regulating mRNA stability is one of the essential mechanisms in gene expression. In order to identify genes from Escherichia coli whole genome whose expression is effectively modulated during the process of mRNA decay, we previously performed differential display-PCR as the first step. In the screening, it was suggested that two mRNAs from the histidine kinase genes, narX and yojN, in a two-component signal transduction system, were extremely unstable. In this study we analysed the stability of sensory kinase mRNAs, e.g. arcB, barA, rcsC, narQ, narX and evgS mRNA. RESULTS: The cellular level of the histidine kinase mRNAs was very low and the mRNAs were rapidly degraded in wild-type cells cultured at 37 degrees C in LB medium. Additional experiments using RNase E deficient cells indicated that the mRNAs existed abundantly and expressed a prolonged half-life in the cells. Monocistronic transcripts of the cognate response regulator genes, arcA, rcsB, narP and narL have a half-life of 1.5-3.4 min. CONCLUSIONS: mRNAs of the six histidine kinase genes in E. coli are synthesized efficiently, but rapidly degraded in wild-type cells. PMID- 12581160 TI - MDG1/ERdj4, an ER-resident DnaJ family member, suppresses cell death induced by ER stress. AB - BACKGROUND: Alterations in homeostasis after various cellular stresses, which prevent protein folding and cause an accumulation of misfolding or malfolding proteins in the endoplasmic reticulum (ER), have the potential to induce cellular damage, and are therefore a type of 'ER stress.' To understand the molecular events or cascades underlying the ER stress response regulated by gene transcription and mediated by stress transducers, it is crucial to identify the molecules induced during ER stress and to analyse the roles of these genes. RESULTS: We identified MDG1/ERdj4, a member of the DnaJ protein family, as an inducible gene during ER stress. MDG1/ERdj4 contains the J domain, which is essential for interacting with Hsp70s, at the N-terminal portion and just at the back of the transmembrane domain. Its trypsin digestion and glycosylation of a chimeric protein composed of MDG1/ERdj4 fused with the extracellular domain of the amyloid precursor protein at its C-terminus, showed that its C-terminal portion containing the J domain could be orientated to the ER lumen. Over expression of it inhibited the cell death induced by ER stress. In contrast, its mutants with the J domain deleted showed no protective effects against cell death. CONCLUSIONS: MDG1/ERdj4 may play roles in stabilizing GRP78/BiP binding to unfolded substrate proteins in a J domain-dependent manner and prevent the accumulation of unfolded proteins in the ER, consequently protecting cells from ER stress. PMID- 12581161 TI - Dopamine gating of forebrain neural ensembles. AB - Dopamine may exert different actions depending on a number of factors. A common view is that D1 receptors may be responsible for excitatory actions whereas D2 receptors are involved in inhibitory actions. However, this position cannot be reconciled with several findings indicating otherwise. The role of dopamine on forebrain neural ensembles may be better understood in the light of functional states of the system. Pyramidal cortical neurons and striatal medium spiny neurons alternate between two membrane potential states ('up' and 'down') that could shape dopamine actions. It is proposed that D1 receptors can act as state stabilizers by sustaining up states and thereby facilitating plasticity mechanisms by providing postsynaptic depolarization and increasing NMDA function. In this way, dopamine can sustain activity in depolarized units. This action is accompanied by a decrease in cell firing (perhaps mediated by D2 receptors), which renders the cells responsive only to strong stimuli. The result would be a net increase in signal-to-noise ratio in a selected assembly of neurons. PMID- 12581162 TI - Odorant specificity of three oscillations and the DC signal in the turtle olfactory bulb. AB - The odour-induced population response in the in vivo turtle (Terepene sp.) olfactory bulb consists of three oscillatory components (rostral, middle and caudal) that ride on top of a DC signal. In an initial step to determine the functional role of these four signals, we compared the signals elicited by different odorants. Most experiments compared isoamyl acetate and cineole, odorants which have very different maps of input to olfactory bulb glomeruli in the turtle and a different perceptual quality for humans. We found substantial differences in the response to the two odours in the rise-time of the DC signal and in the latency of the middle oscillation. The rate of rise for cineole was twice as fast as that for isoamyl acetate. Similarly, the latency for the middle oscillation was about twice as long for isoamyl acetate as it was for cineole. On the other hand, a number of characteristics of the signals were not substantially different for the two odorants. These included the latency of the rostral and caudal oscillation, the frequency and envelope of all three oscillations and their locations and spatial extents. A smaller number of experiments were carried out with hexanone and hexanal; the oscillations elicited by these odorants did not appear to be different from those elicited by isoamyl acetate and cineole. Qualitative differences between the oscillations in the turtle and those in two invertebrate phyla suggest that different odour processing strategies may be used. PMID- 12581163 TI - Endogenous and exogenous dopamine presynaptically inhibits glutamatergic reticulospinal transmission via an action of D2-receptors on N-type Ca2+ channels. AB - In this study, the effects of exogenously applied and endogenously released dopamine (DA), a powerful modulator of the lamprey locomotor network, are examined on excitatory glutamatergic synaptic transmission between reticulospinal axons and spinal neurons. Bath application of DA (1-50 micro m) reduced the amplitude of monosynaptic reticulospinal-evoked glutamatergic excitatory postsynaptic potentials (EPSPs). The effect of DA was blocked by the D2-receptor antagonist eticlopride, and mimicked by the selective D2-receptor agonist 2,10,11 trihydroxy-N-propyl-noraporphine hydrobromide (TNPA). Bath application of the DA reuptake blocker bupropion, which increases the extracellular level of dopamine, also reduced the monosynaptic EPSP amplitude. This effect was also blocked by the D2-receptor antagonist eticlopride. To investigate if the action of DA was exerted at the presynaptic level, the reticulospinal axon action potentials were prolonged by administering K+ channel antagonists while blocking l-type Ca2+ channels. A remaining Ca2+ component, mainly dependent on N and P/Q channels, was depressed by DA. When DA (25-50 micro m) was applied in the presence of omega conotoxin GVIA, a toxin specific for N-type Ca2+ channels, it failed to affect the monosynaptic EPSP amplitude. DA did not affect the response to extracellularly ejected d-glutamate, the postsynaptic membrane potential, or the electrical component of the EPSPs. DA thus acts at the presynaptic level to modulate reticulospinal transmission. PMID- 12581164 TI - The effects of nitric oxide on magnocellular neurons could involve multiple indirect cyclic GMP-dependent pathways. AB - Nitric oxide (NO) is known to regulate the release of arginine-vasopressin (AVP) and oxytocin (OT) by the paraventricular nucleus (PVN) and the supraoptic nucleus (SON). The aim of the current study was to identify in these nuclei the NO producing neurons and the NO-receptive cells in mice. The determination of NO synthesizing neurons was performed by double immunohistochemistry for the neuronal form of NO synthase (NOS), and AVP or OT. Besides, we visualized the NO receptive cells by detecting cyclic GMP (cGMP), the major second messenger for NO, by immunohistochemistry on hypothalamus slices. Neuronal NOS was exclusively colocalized with OT in the PVN and the SON, suggesting that NO is mainly synthesized by oxytocinergic neurons in mice. By contrast, cGMP was not observed in magnocellular neurons, but in GABA-, tyrosine hydroxylase- and glutamate positive fibers, as well as in GFAP-stained cells. The cGMP-immunostaining was abolished by incubating brain slices with a NOS inhibitor (L-NAME). Consequently, we provide the first evidence that NO could regulate the release of AVP and OT indirectly by modulating the activity of the main afferents to magnocellular neurons rather than by acting directly on magnocellular neurons. Moreover, both the NADPH-diaphorase activity and the mean intensity of cGMP-immunofluorescence were increased in monoamine oxidase A knock-out mice (Tg8) compared to control mice (C3H) in both nuclei. This suggests that monoamines could enhance the production of NO, contributing by this way to the fine regulation of AVP and OT release and synthesis. PMID- 12581165 TI - A radialization factor in normal cortical plate restores disorganized radial glia and disrupted migration in a model of cortical dysplasia. AB - Treatment of pregnant ferrets on embryonic day 24 (E24) with the antimitotic methylazoxy methanol (MAM) leads to a specific constellation of effects in newborn kits, which include a very thin and poorly laminated neocortex, disruption of radial glial cell morphology with early differentiation into astrocytes, and abnormal positioning of Cajal-Retzius cells. We suggest that MAM treatment on E24 results in this model of cortical dysplasia by eliminating a population of cells that produce a factor capable of maintaining radial glia in their normal morphology. The abnormal radial glia, either alone or in combination with other abnormal features, are likely to prevent proper migration into the cortical plate. To test the possibility that normal cortex can provide the missing substance that influences radial glia, slices of E24 MAM-treated cortex were removed at postnatal day 0 (P0) and cultured adjacent to explants of P0 normal cortical plate. By labelling a small number of cells with injections of fluorescent dextrans into the cultured slices, we found that abnormal radial glia in MAM treated slices cocultured adjacent to normal cortical plate were restored toward normal, in comparison to E24 MAM treated slices cultured alone and in other control conditions. We also found that abnormally positioned Cajal-Retzius cells move into the marginal zone and that neurons are able to migrate into the cortical plate more effectively in the coculture condition. These data indicate that normal cortical plate of ferrets contains a factor causing radial glia to maintain their elongated morphology; the improved position of radial glia encourages repositioning of Cajal-Retzius cells and improved neuronal migration into the cortical plate. PMID- 12581166 TI - Differential galanin receptor-1 and galanin expression by 5-HT neurons in dorsal raphe nucleus of rat and mouse: evidence for species-dependent modulation of serotonin transmission. AB - Galanin and galanin receptors are widely expressed by neurons in rat brain that either synthesize/release and/or are responsive to, classical transmitters such as gamma-aminobutyric acid, acetylcholine, noradrenaline, histamine, dopamine and serotonin (5-hydroxytryptamine, 5-HT). The dorsal raphe nucleus (DRN) contains approximately 50% of the 5-HT neurons in the rat brain and a high percentage of these cells coexpress galanin and are responsive to exogenous galanin in vitro. However, the precise identity of the galanin receptor(s) present on these 5-HT neurons has not been previously established. Thus, the current study used a polyclonal antibody for the galanin receptor-1 (GalR1) to examine the possible expression of this receptor within the DRN of the rat and for comparative purposes also in the mouse. In the rat, intense GalR1-immunoreactivity (IR) was detected in a substantial population of 5-HT-immunoreactive neurons in the DRN, with prominent receptor immunostaining associated with soma and proximal dendrites. GalR1-IR was also observed in many cells within the adjacent median raphe nucleus. In mouse DRN, neurons exhibited similar levels and distribution of 5-HT-IR to that in the rat, but GalR1-IR was undetectable. Consistent with this, galanin and GalR1 mRNA were also undetectable in mouse DRN by in situ hybridization histochemistry, despite the detection of GalR1 mRNA (and GalR1-IR) in adjacent cells in the periaqueductal grey and other midbrain areas. 5-HT neuron activity in the DRN is primarily regulated via 5-HT1A autoreceptors, via inhibition of adenylate cyclase and activation of inward-rectifying K+ channels. Notably, the GalR1 receptor subtype signals via identical mechanisms and our findings establish that galanin modulates 5-HT neuron activity in the DRN of the rat via GalR1 (auto)receptors. However, these studies also identify important species differences in the relationship between midbrain galanin and 5-HT systems, which should prompt further investigations in relation to comparative human neurochemistry and which have implications for studies of animal models of relevant neurological conditions such as stress, anxiety and depression. PMID- 12581167 TI - Rat strain differences in peripheral and central serotonin transporter protein expression and function. AB - Female Fischer 344 (F344) rats have been shown to display increased serotonin transporter (5-HTT) gene expression in the dorsal raphe, compared to female Lewis (LEW) rats. Herein, we explored, by means of synaptosomal preparations and in vivo microdialysis, whether central, but also peripheral, 5-HTT protein expression/function differ between strains. Midbrain and hippocampal [3H]paroxetine binding at the 5-HTT and hippocampal [3H]serotonin (5-HT) reuptake were increased in male and female F344 rats, compared to their LEW counterparts, these strain differences being observed both in rats of commercial origin and in homebred rats. Moreover, in homebred rats, it was found that these strain differences extended to blood platelet 5-HTT protein expression and function. Saturation studies of midbrain and hippocampal [3H]paroxetine binding at the 5 HTT, and hippocampal and blood platelet [3H]5-HT reuptake, also revealed significant strain differences in Bmax and Vmax values. Although F344 and LEW rats differ in the activity of the hypothalamo-pituitary-adrenal (HPA) axis, manipulations of that axis revealed that the strain differences in hippocampal [3H]paroxetine binding at 5-HTTs and [3H]5-HT reuptake were not accounted for by corticosteroids. Hippocampal extracellular 5-HT levels were reduced in F344 rats, compared to LEW rats, with the relative, but not the absolute, increase in extracellular 5-HT elicited by the local administration of citalopram being larger in F344 rats. Because the aforementioned strain differences did not lie in the coding sequences of the 5-HTT gene, our results open the promising hypothesis that F344 and LEW strains model functional polymorphisms in the promoter region of the human 5-HTT gene. PMID- 12581168 TI - AMPA/kainate and NMDA-like glutamate receptors at the chromatophore neuromuscular junction of the squid: role in synaptic transmission and skin patterning. AB - Glutamate receptor types were examined at the chromatophore synapses of the squids Alloteuthis subulata and Loligo vulgaris, where nerve-induced muscle contraction causes chromatophore expansion. Immunoblotting with antibody raised against a squid AMPA receptor (sGluR) demonstrated that AMPA/kainate receptors are present in squid skin. Application of l-glutamate evoked chromatophore muscle contractions in both ventral and dorsal skins, while NMDA was only active on a subpopulation of dorsal chromatophores. In dorsal skin, neurotransmission was partly blocked by either AMPA/kainate receptor antagonists (CNQX and DNQX) or NMDA receptor antagonists (AP-5 and MK-801) or completely blocked by simultaneous application of both classes of antagonists. In isolated muscle fibres, ionophoretic application of l-glutamate evoked fast inward CNQX- and DNQX sensitive currents with reversal potentials around +14 mV and a high conductance to Na+. In fibres from dorsal skin only, a slower outward glutamate-sensitive current appeared at positive holding potentials. At negative potentials, currents were potentiated by glycine or by removing external Mg2+ and were blocked by AP-5 and MK-801. Glutamate caused a fast, followed by a slow, transient increase in cytoplasmic Ca2+. The slow component was increased in amplitude and duration by glycine or by lowering external Mg2+ and decreased by AP-5 and MK-801. In cells from ventral skin, no 'NMDA-like responses' were detected. Thus, while AMPA/kainate receptors mediated fast excitatory synaptic transmission and rapid colour change over the whole skin, activation of both AMPA/kainate and NMDA-like receptors in a subpopulation of dorsal chromatophores prolonged the postsynaptically evoked Ca2+ elevation causing temporally extended colour displays with behavioural significance. PMID- 12581169 TI - Disabled-1 mRNA and protein expression in developing human cortex. AB - Disabled-1 (Dab1) forms part of the Reelin-Dab1 signalling pathway that controls neuronal positioning during brain development; Dab1 deficiency gives rise to a reeler-like inversion of cortical layers. To establish a timetable of Dab1 expression in developing human brain, Dab1 mRNA and protein expression were studied in prenatal human cortex. The earliest Dab1 signal was detected at 7 gestational weeks (GW), the stage of transition from preplate to cortical plate, suggesting a role of the Reelin-Dab1 signalling pathway in preplate partition. From 12 to 20 GW, the period of maximum cortical migration, Dab1 expression was prominent in the upper tiers of the cortical plate, to decline after midgestation. Radially orientated apical dendrites of Dab1-expressing neurons indicated a predominant pyramidal phenotype. Pyramidal cells in hippocampus and entorhinal cortex displayed a more protracted time of Dab1 expression compared to neocortex. In addition, at later stages (18-25 GW), Dab1 was also expressed in large neurons scattered throughout intermediate zone and subplate. From 14 to 22 GW, particularly high levels of Dab1 mRNA and protein were observed in cells of the ventricular/subventricular zone displaying the morphology of radial glia. The partial colocalization of vimentin and Dab1 in cells of the ventricular zone supported a radial glia phenotype. The concentration of Dab1 protein in ventricular endfeet and initial portions of radial processes of ventricular-zone cells points to a possible involvement of Dab1 in neurogenesis. Furthermore, a subset of Cajal-Retzius cells in the marginal zone colocalized Dab1 and Reelin, and may thus represent a novel target of the Reelin-Dab1 signalling pathway. PMID- 12581170 TI - Modulation of spontaneous and evoked EPSCs and IPSCs in optic lobe neurons of cuttlefish Sepia officinalis by the neuropeptide FMRF-amide. AB - The effects of the neuropeptide FMRFa on spontaneous excitatory postsynaptic currents (sEPSCs) and inhibitory postsynaptic currents (sIPSCs), as well as on evoked EPSCs and IPSCs, in two types of neurons within the central optic lobe of cuttlefish were examined using the whole-cell voltage-clamp technique. FMRFa (1 10 micro m) did not affect cell membrane resting potentials, but reversibly reduced both the frequency and amplitude of sEPSCs in neurons within the medulla region of the optic lobe while increasing the frequency and amplitude of their sIPSCs. For centrifugal neurons in the inner granule cell layer of the optic lobe, FMRFa (1-10 micro m) decreased both the frequency and amplitude of sEPSCs. In the presence of tetrodotoxin (0.5 micro m), neither the interevent interval, nor amplitude distributions of the miniature EPSCs or the miniature IPSCs, were affected by FMRFa, implying a presynaptic action of FMRFa on the optic lobe neurons. Bath application of the neuropeptide also abolished or reduced in amplitude the evoked EPSCs and increased the amplitude of evoked IPSCs in optic lobe neurons, showing that FMRFa induced similar effects on evoked as on spontaneous postsynaptic currents. These results demonstrate the complex range of modulatory effects FMRFa can have within central nervous system circuits. PMID- 12581171 TI - Neurons with distinctive firing patterns, morphology and distribution in laminae V-VII of the neonatal rat lumbar spinal cord. AB - It is generally accepted that neurons in the ventral spinal grey matter, a substantial proportion of which can be regarded as constituents of the spinal motor apparatus, receive and integrate synaptic inputs arising from various peripheral, spinal and supraspinal sources. Thus, a profound knowledge concerning the integrative properties of interneurons in the spinal ventral grey matter appears to be essential for a fair understanding of operational principles of spinal motor neural assemblies. Using the whole cell patch clamp configuration in a correlative physiological and morphological experimental approach, here we demonstrate that the intrinsic membrane properties of neurons vary widely in laminae V-VII of the ventral grey matter of the neonatal rat lumbar spinal cord. Based on their firing patterns in response to depolarizing current steps, we have classified the recorded neurons into four categories: 'phasic', 'repetitive', 'single' and 'slow'. Neurons with firing properties characteristic of the 'phasic', 'repetitive' and 'single' cells have previously been reported also in the superficial and deep spinal dorsal horn, but this is the first account in the literature in which 'slow' neurons have been recovered and described in the spinal cord. The physiological heterogeneity in conjunction with the morphological correlation and distribution of neurons argues that different components of motor neural assemblies in the spinal ventral grey matter possess different signal processing characteristics. PMID- 12581172 TI - Effects of insulin-like growth factor I on climbing fibre synapse elimination during cerebellar development. AB - Functional neural circuit formation includes the process by which redundant synaptic connections formed earlier during development are subsequently eliminated. We report that insulin-like growth factor I (IGF-I) is a candidate factor that influences the developmental transition from multiple to mono innervation of cerebellar Purkinje cells (PCs) by climbing fibres (CFs). Continuous local application of exogenous IGF-I to the mouse cerebellum by means of ethylene-vinyl acetate copolymer (Elvax) significantly increased the degree of multiple CF innervation, when the IGF-I containing Elvax was implanted at postnatal day 8 (P8). In contrast, the IGF-I application starting at P12 had no effect on CF innervation. Conversely, continuous local application of antisera against IGF-I and its receptor significantly decreased the degree of multiple CF innervation when the application started at P8. We found that chronic treatment of exogenous IGF-I from P8 significantly enhanced the CF-mediated excitatory postsynaptic currents (CF-EPSCs). This effect was manifest for the smaller CF EPSCs but not for the largest CF-EPSC of the multiple-innervated PCs. Conversely, chronic application of antisera from P8 caused attenuation of the largest CF EPSCs. Other parameters for basic synaptic functions and cerebellar morphology were largely normal after the IGF-I or antisera treatment. These results suggest that IGF-I enhances the strength of developing CF synapses and may promote their survival, whereas the shortage of IGF-I impairs the development of CF synapses and, as a result, may facilitate their elimination. Thus, IGF-I is a potentially important factor among various signalling molecules that can influence CF synapse elimination during cerebellar development. PMID- 12581173 TI - Presynaptic inhibition of Schaffer collateral synapses by stimulation of hippocampal cholinergic afferent fibres. AB - It has been known for decades that muscarinic agonists presynaptically inhibit Schaffer collateral synapses contacting hippocampal CA1 pyramidal neurons. However, a demonstration of the inhibition of Schaffer collateral synapses induced by acetylcholine released by cholinergic hippocampal afferents is lacking. We present original results showing that electrical stimulation at the stratum oriens/alveus with brief stimulus trains inhibited excitatory postsynaptic currents evoked by stimulation of Schaffer collaterals in CA1 pyramidal neurons of rat hippocampal slices. The increased paired-pulse facilitation and the changes in the variance of excitatory postsynaptic current amplitude that paralleled the inhibition suggest that it was mediated presynaptically. The effects of oriens/alveus stimulation were inhibited by atropine, and blocking nicotinic receptors with methyllycaconitine was ineffective, suggesting that the inhibition was mediated via the activation of presynaptic muscarinic receptors. The results provide a novel demonstration of the presynaptic inhibition of glutamatergic neurotransmission by cholinergic fibres in the hippocampus, implying that afferent cholinergic fibres regulate the strength of excitatory synaptic transmission. PMID- 12581174 TI - Prefrontal and agranular cingulate projections to the dorsal premotor areas F2 and F7 in the macaque monkey. AB - The superior sector of Brodmann area 6 (dorsal premotor cortex, PMd) of the macaque monkey consists of a rostral and a caudal architectonic area referred to as F7 and F2, respectively. The aim of this study was to define the origin of prefrontal and agranular cingulate afferents to F7 and F2, in the light of functional and hodological evidence showing that these areas do not appear to be functionally homogeneous. Different sectors of F7 and F2 were injected with neural tracers in seven monkeys and the retrograde labelling was qualitatively and quantitatively analysed. The dorsorostral part of F7 (supplementary eye field, F7-SEF) was found to be a target of strong afferents from the frontal eye field (FEF), from the dorsolateral prefrontal regions located dorsally (DLPFd) and ventrally (DLPFv) to the principal sulcus and from cingulate areas 24a, 24b and 24c. In contrast, the remaining part of F7 (F7-non SEF) is only a target of the strong afferents from DLPFd. Finally, the ventrorostral part of F2 (F2vr), but not the F2 sector located around the superior precentral dimple (F2d), receives a minor, but significant, input from DLPFd and a relatively strong input from the cingulate gyrus (areas 24a and 24b) and area 24d. Present data provide strong hodological support in favour of the idea that areas F7 and F2 are formed by two functionally distinct sectors. PMID- 12581175 TI - Differential routing of coexisting neuropeptides in vasopressin neurons. AB - The functional implications of intraneuronal coexistence of different neuropeptides depend on their respective targeting to release sites. In the rat hypothalamic magnocellular neurons, we investigated a possible differential routing of the coexpressed galanin and vasopressin. The respective location of proteins and messengers was assessed with double immunogold and in situ hybridization combining confocal and electron microscope analysis. The various populations of labelled granules were quantitatively compared in three subcellular compartments: perikarya, local processes and posthypophyseal nerve endings. Three subpopulations of granules were detected in all three compartments, but their respective amount showed significant differences. Galanin alone was immunolocalized in some secretory granules, vasopressin alone in others, and both peptides in a third subpopulation of granules. The major part of the granules containing vasopressin, either alone or in association with galanin, is found in neurohypophyseal nerve endings. In contrast, galanin single-labelled granules represent the most abundant population in dendritic processes, while double-labelled granules are more numerous in perikarya. This indicates a preferential distribution of the two peptides in the different compartments of magnocellular neurons. Furthermore, galanin and vasopressin messenger RNAs were detected at different domains of the endoplasmic reticulum, suggesting that translation might also occur at different locations, thus leading to partial segregation of galanin and vasopressin cargoes between two populations of secretory granules. The present study provides, for the first time in mammals, evidence suggesting that galanin and vasopressin are only partly copackaged and undergo a preferential targeting toward dendrites or neurohypophysis, suggesting different functions, autocrine/paracrine and endocrine, respectively. PMID- 12581176 TI - Behavioural sensitization and enhanced dopamine response in the nucleus accumbens after intravenous cocaine self-administration in mice. AB - The behavioural effects of cocaine are enhanced in animals with a prior history of repeated cocaine administration. This phenomenon, referred to as sensitization, is also associated with an increase in cocaine-evoked extracellular dopamine levels in the nucleus accumbens. Behavioural and neurochemical sensitization has been demonstrated in rats with a prior history of cocaine self-administration and in those that had received experimenter administered cocaine. Although it is clear that the repeated non-contingent administration also results in behavioural sensitization in the mouse, the issue of whether behavioural and neurochemical sensitization also occur in this species following intravenous cocaine self-administration has not been assessed. The present study used the technique of in vivo microdialysis in conjunction with operant self-administration to characterize cocaine-evoked locomotor activity and dopamine levels in the nucleus accumbens in mice with a prior history of intravenous cocaine self-administration or those that had received yoked infusions of cocaine. Mice that had received contingent or non-contingent infusions of cocaine exhibited an enhanced behavioural response to cocaine and increased cocaine-evoked dopamine levels in the nucleus accumbens. There was no difference between groups in the magnitude of this effect. Prior exposure to cocaine did not modify baseline dopamine levels in the nucleus accumbens. These data demonstrate that mice with previous cocaine self-administration experience show an enhanced behavioural and dopamine response to cocaine in the nucleus accumbens. Furthermore, control over cocaine infusion does not significantly alter the magnitude of the sensitized behavioural and presynaptic dopamine responses observed in response to a challenge dose of cocaine. PMID- 12581177 TI - Intrathecally applied flurbiprofen produces an endocannabinoid-dependent antinociception in the rat formalin test. AB - It is generally accepted that the phospholipase-A2-cyclooxygenase-prostanoids cascade mediates spinal sensitization and hyperalgesia. However, some observations are not in line with this hypothesis. The aim of the present work was to investigate whether different components of this cascade exhibit nociceptive or antinociceptive effects in the rat formalin test. Intrathecal (i.th.) injection of prostaglandin E2 (PGE2) induced a dose-dependent antinociceptive effect on the formalin-induced nociception. Furthermore, thimerosal, which inhibits the reacylation of arachidonic acid thereby enhancing arachidonic acid levels, had an antinociceptive effect rather than the expected pronociceptive effect when given i.th. While the phospholipase A2 inhibitor methyl arachidonyl fluorophosphonate (MAFP; i.th.) had a significant antinociceptive effect, its analogue palmitoyl trifluoromethyl ketone (PTFMK; i.th.) had no significant effect on the formalin-induced nociception. However, MAFP, but not PTFMK, showed a cannabinoid CB1 agonistic effect as shown by the inhibition of electrically evoked contractions of the vas deferens isolated from CB1 wild-type mice but not of that from CB1 knockout mice. The antinociceptive effect of MAFP was completely reversed by the CB1 receptor antagonist AM-251 (i.th.), thus attributing such effect to its CB1 agonistic effect. Moreover, the antinociceptive effect of the cyclooxygenase inhibitor, flurbiprofen (i.th.) was reversed by the co-administration of AM-251, but not by PGE2. Finally. the combination of phenylmethylsulfonyl fluoride (PMSF; intraperitoneal), which inhibits the degradation of anandamide through the inhibition of fatty acid amidohydrolase, with thimerosal (i.th.) produced a profound CB1-dependent antinociception. The present results show that endocannabinoids play a major role in mediating flurbiprofen-induced antinociception at the spinal level. PMID- 12581178 TI - Morphine withdrawal-induced abnormalities in the VTA: confocal laser scanning microscopy. AB - Morphine withdrawal is characterized by functional alterations at the level of the ventrotegmental area. We investigated the effects of chronic morphine administration and withdrawal on the morphological properties of immuno-labelled tyrosine hydroxylase-positive neurons of the rat ventrotegmental area with a confocal laser scanning microscope. Morphological evaluation revealed a reduction in the area and perimeter of tyrosine hydroxylase-positive somata in morphine withdrawn rats. Conversely, the number of cells per field was found to have increased in the naloxone group. Collectively, the present results indicate that withdrawal from a chronic morphine treatment, and not chronic morphine per se, modifies cellular morphology of tyrosine hydroxylase-positive, presumably dopamine-containing, neurons of the rat VTA. This is consistent with the idea that withdrawal from morphine alters functioning of the mesolimbic dopamine system and provides a direct morphological correlate for the functional abnormalities typical of morphine withdrawal. PMID- 12581179 TI - Modulatory action of acetylcholine on striatal neurons: microiontophoretic study in awake, unrestrained rats. AB - Cholinergic interneurons innervate virtually all medium spiny striatal cells, but the relevance of this input in regulating the activity and afferent responsiveness of these cells remains unclear. Studies in anaesthetized animals and slice preparations have shown that iontophoretic acetylcholine (ACh) either weakly excites or inhibits striatal neurons. These differential responses may reflect cholinergic receptor heterogeneity but may be also related to the different activity states of recorded units and different afferent inputs specific in each preparation. Single-unit recording was combined with iontophoresis in awake, unrestrained rats to examine the effects of ACh and selective muscarinic (oxotremorine M or Oxo-M) and nicotinic agonists (nicotine or NIC) on dorsal and ventral striatal neurons. These effects were tested on naturally silent, spontaneously active and glutamate-stimulated units. We found that iontophoretic ACh primarily inhibited spontaneously active and glutamate stimulated units; the direction of the ACh response, however, was dependent on the firing rate. The effects of ACh were generally mimicked by Oxo-M and, surprisingly, by NIC, which is known to excite units in most central structures, including striatal neurons in anaesthetized preparation. Given that NIC receptors are absent on striatal cells but located primarily on dopamine terminals, we assessed the effects of NIC after complete blockade of dopamine receptors induced by systemic administration of a mixture of D1 and D2 antagonists. During dopamine receptor blockade the number of NIC-induced inhibitions dramatically decreased and NIC had mainly excitatory effects on striatal neurons. Thus, our data suggest that under physiologically relevant conditions ACh acts as a state-dependent neuromodulator, and its action involves not only postsynaptic but also presynaptic cholinoreceptors located on dopamine- and glutamate-containing terminals. PMID- 12581180 TI - Induction of bilateral plasticity in sensory cortical maps by small unilateral cortical infarcts in rats. AB - Behavioural impairments caused by brain lesions show a considerable, though often incomplete, recovery. It is hypothesized that cortical and subcortical plasticity of sensory representations contribute to this recovery. In the hindpaw representation of somatosensory cortex of adult rats we investigated the effects of focal unilateral cortical lesions on remote areas. Cortical lesions with a diameter of approximately 2 mm were induced in the parietal cortex by photothrombosis with the photosensitive dye Rose Bengal. Subsequently, animals were kept in standard cages for 7 days. On day seven, animals were anaesthetized and cutaneous receptive fields in the cortical hindpaw representations ipsi- and contralateral to the lesion were constructed from extracellular recordings of neurons in layer IV using glass microelectrodes. Receptive fields in the lesioned animals were compared to receptive fields measured in nonlesioned animals serving as controls. Quantitative analysis of receptive fields revealed a significant increase in size in the lesioned animals. This doubling in receptive field size was observed equally in the hemispheres ipsi- and contralateral to the lesion. The results indicate that the functional consequences of restricted cortical lesions are not limited to the area surrounding the lesion, but affect the cortical maps on the contralateral, nonlesioned hemisphere. PMID- 12581181 TI - Fos imaging reveals ageing-related changes in hippocampal response to radial maze discrimination testing in mice. AB - A two-stage radial arm maze (RAM) task has been designed recently to demonstrate a specific age-related memory deficit in mice. It highlights the contrast between normal and deficient memory expression in a spatial discrimination problem depending on how to-be discriminated arms were presented to the animal. This specific deficit has been interpreted as a preferential loss in a relational/declarative form of memory, thereby implying an underlying hippocampal dysfunction. To test this hypothesis, neuronal activation measured by Fos immunostaining was compared in aged (21-23 months) and adult (4-6 months) mice trained in the aforementioned task and killed after a retention session consisting in age-insensitive probe trials, performed 6 days later (6-day RAM). Two comparison conditions were included: (i) repeated locomotor training on a treadmill (TM); (ii) the same RAM training, except for the use of a longer (30 days instead of six) retention interval (30-day RAM). Although all RAM groups displayed similar levels of performance at the end of the experiment, immediately before the mice were killed, significant between-group differences in brain activation were observed. In adult mice, 6-day RAM testing was associated with greater septal and hippocampal (CA1, CA3, DG) Fos expression than the TM condition. Lengthening the retention interval from 6 days to 30 days resulted in a significant decrease in RAM testing-induced Fos expression in most of the septo hippocampal regions. With respect to adult mice, aged mice displayed reduced Fos expression (except for DG) and a lack of interrelationships between levels of Fos produced in each of the SH regions, in the 6-day RAM testing condition. Conversely, there was no effect of ageing on Fos expression associated with either TM training or 30-day RAM testing. These results are interpreted as reflecting age- (or time-) related alterations in recruiting of brain structures that underlie a relational/declarative form of memory expression. PMID- 12581183 TI - Impairments in visual discrimination after perirhinal cortex lesions: testing 'declarative' vs. 'perceptual-mnemonic' views of perirhinal cortex function. AB - Two experiments tested the predictions of 'declarative' vs. 'perceptual-mnemonic' views of perirhinal cortex function. The former view predicts that perirhinal cortex lesions should impair rapidly learned, but not more slowly learned, visual discriminations, whereas the latter view predicts that impairments should be related not to speed of learning but to perceptual factors. It was found that monkeys with perirhinal cortex lesions were impaired in the acquisition and performance of slowly learned, perceptually difficult greyscale picture discriminations, but were not impaired in the acquisition of rapidly learned, perceptually easier discriminations. In addition, these same monkeys were not impaired in the acquisition or performance of difficult colour or size discriminations, indicating that the observed pattern of impairments was not due to ceiling effects or difficulty per se. These findings, taken together, are consistent with the 'perceptual-mnemonic' view that the perirhinal cortex is involved in both perception and memory, but are not consistent with the 'declarative' view that the perirhinal cortex is important exclusively for declarative memory, having little or no role in perception. Moreover, the results are consistent with the more specific proposal that the perirhinal cortex contributes to the solution of complex visual discriminations with a high degree of 'feature ambiguity', a property of visual discrimination problems that can emerge when features of an object are rewarded when part of one object, but not when part of another. These and other recent findings suggest the need for a revision of prevailing views regarding the neural organization of perception and memory. PMID- 12581184 TI - Time course of striatal DeltaFosB-like immunoreactivity and prodynorphin mRNA levels after discontinuation of chronic dopaminomimetic treatment. AB - DeltaFosB-like proteins are particularly stable transcription factors that accumulate in the brain in response to chronic perturbations. In this study we have compared the time-course of striatal FosB/DeltaFosB-like immunoreactivity and prodynorphin mRNA expression after discontinuation of chronic cocaine treatment to intact rats and chronic L-DOPA treatment to unilaterally 6 hydroxydopamine (6-OHDA) lesioned rats. The animals were killed between 3 h and 16 days after the last drug injection. In both treatment paradigms, the drug induced FosB/DeltaFosB immunoreactivity remained significantly elevated in the caudate putamen even at the longest withdrawal period examined. The concomitant upregulation of prodynorphin mRNA, a target of DeltaFosB, paralleled the time course of DeltaFosB-like immunoreactivity in the 6-OHDA-lesion/L-DOPA model, but was more transient in animals treated with cocaine. These results suggest that DeltaFosB-like proteins have exceptional in vivo stability. In the dopamine denervated striatum, these proteins may exert sustained effects on the expression of their target genes long after discontinuation of L-DOPA pharmacotherapy. PMID- 12581182 TI - Sex differences in anxiety, sensorimotor gating and expression of the alpha4 subunit of the GABAA receptor in the amygdala after progesterone withdrawal. AB - In a progesterone withdrawal (PWD) model of premenstrual anxiety, we have previously demonstrated that increased hippocampal expression of the alpha4 subunit of the GABAA receptor (GABAA-R) is closely associated with higher anxiety levels in the elevated plus maze. However, several studies indicate that sex differences in regulation of the GABAA-R in specific brain regions may be an important factor in the observed gender differences in mood disorders. Thus, we investigated possible sex differences in GABAA-R subunit expression and anxiety during PWD. To this end, we utilized the acoustic startle response (ASR) to assess anxiety levels in male and female rats undergoing PWD as the ASR is also applicable to the assessment of human anxiety responses. We also investigated GABAA-R alpha4 subunit expression in the amygdala, as the amygdala directly regulates the primary startle circuit. Female rats exhibited a greater ASR during PWD than controls, indicating higher levels of anxiety and arousal. In contrast, male rats undergoing PWD did not demonstrate an increased ASR. The sex differences in the ASR were paralleled by sex differences in the expression of the GABAA-R alpha4 subunit in the amygdala such that alpha4 subunit expression was up-regulated in females during PWD whereas alpha4 levels in males undergoing PWD were not altered relative to controls. These findings might have implications regarding gender differences in human mood disorders and the aetiology of premenstrual anxiety. PMID- 12581185 TI - Cognitive control processes during an anticipated switch of task. AB - For successful negotiation of our environment, humans must be readily able to switch from one task to another. This ability relies on 'executive control' processes and despite extensive efforts to detail the nature of these processes, there is little consensus as to how the brain achieves this critical function. Behavioural studies show that as subjects are given more time to prepare to switch task, performance improves; yet even with the longest preparation intervals, there remains an ineradicable performance cost on switch trials. As such, some elements of the switching process must wait until the stimulus to be acted upon has actually been presented. Here, using the methods of high-density mapping of brain potentials, we show that early visual processes are substantially different on switch trials than on later trials. Our data show that while there is clearly a degree of preparatory processing that occurs prior to a predictable switch of task, some elements of switching are only achieved after the switch stimulus has been presented. Our findings are discussed in the context of a new model of executive control processes that suggests that preparing to switch task may not be a separate (control) process per se, but rather, the beginning of a competition between the potentially relevant tasks, a competition that is ultimately resolved during the switch trial. PMID- 12581186 TI - Anti-HPA-1a-mediated platelet phagocytosis by monocytes in vitro and its inhibition by Fc gamma receptor (FcgammaR) reactive reagents. AB - The study was undertaken to delineate mechanisms of platelet destruction by phagocytosis during fetal/neonatal alloimmune thrombocytopenia (FAIT/NAIT) because of maternal antibodies against human platelet antigen 1a (HPA-1a). By employing a platelet phagocytosis assay based on the ORPEGEN flow cytometric bacterial phagocytosis test, we measured monocyte ingestion of platelets mediated by anti-HPA-1a antibodies. Moreover, we tested, as potential therapeutic agents, FcgammaR reactive reagents, for their inhibition of this process. Four of six anti-HPA-1a sera tested mediated phagocytosis of HPA-1a-positive platelets in a concentration-dependent manner. Monocyte ingestion of platelets was almost completely inhibited by cytochalasin D. No anti-HPA-1a-mediated phagocytosis was observed with anti-HPA-1a-negative platelets. The humanised anti-FcgammaRI monoclonal antibody H22 at concentrations 1-100 microg/ml, completely inhibited anti-HPA-1a-mediated phagocytosis as did similar concentrations of ivIg. By contrast, a mouse monoclonal anti-FcgammaRII (IV.3, Fab) at 10 microg/ml caused little or no suppression of platelet phagocytosis mediated by two anti-HPA-1 sera. Furthermore, the addition of anti-FcgammaRII (10 microg/ml) to sub-optimal concentrations of H22 did not significantly increase the inhibitory effect of the latter compound. Monomeric IgG (0.1-10 microg/ml) failed to suppress anti-HPA-1 mediated platelet ingestion by the phagocytes, as did anti-FcgammaRIII. To our knowledge this is a rare example of an assay that measures platelet phagocytosis in vitro. The results suggest that FcgammaRI plays a major role in anti-HPA-1a mediated platelet phagocytosis by monocytes while FcgammaRIIa, is of little or minor importance only. Moreover, the findings indicate the use of H22 as an alternative to interavenous Ig (ivIg) in the management of FAIT/NAIT. PMID- 12581187 TI - Distribution and frequency of beta-thalassemia mutations in northwestern and central Greece. AB - OBJECTIVES: Beta-Thalassemia is a common autosomal recessive disorder resulting from over 200 different mutations of the beta-globin genes. The spectrum of beta thalassemia mutations in Greece has been previously described in the population of the capital city of Athens, or in beta-thalassemia patients having transfusion therapy. The aim of the present study was to identify the distribution of the most common beta-thalassemia mutations in the population of northwestern and central Greece. METHODS: The data for this study were derived from a total of 1,130 unrelated subjects including 46 beta-thalassemia major, three beta thalassemia intermedia and 1,081 carriers identified in our antenatal screening program. beta-Thalassemia mutations were identified by ARMS, DGGE and Reverse Dot Blot. RESULTS: The most common mutation, IVS-I-110, is followed, in order of frequency, by the mutations Cd-39, IVS-I-1, IVS-II-1, Cd-6, IVS-I-6, IVS-I-5, IVS II-745, Cd-5 and 44 bp del. IVS-I-110 and Cd-39 frequencies are similar with those found in other Balkan countries. Significant differences in regional distribution were observed. The results showed a clear drift of the distribution of the most frequent IVS-I-110 mutation in the south-north (29.4, 40.0, 44.6 and 61.7%) and the east-west axis (31.8 and 44.6%). CONCLUSIONS: Population screening and prenatal diagnosis are significantly facilitated by these data. Furthermore, the detailed distribution tables of beta-thalassemia mutations are essential for counseling and extraction of genetic diversity estimates for population genetic studies in other inherited disorders. PMID- 12581188 TI - Beta-globin gene cluster haplotypes and HbF levels are not the only modulators of sickle cell disease in Lebanon. AB - Sickle cell disease (SCD) is an inherited autosomal recessive disorder of the beta-globin chain. Despite the fact that all subjects with SCD have the same single base pair mutation, the severity of the clinical and hematological manifestations is extremely variable. This study examined for the first time in Lebanon the correlation between the clinical manifestation of SCD and the beta globin gene haplotypes. The haplotypes of 50 patients diagnosed with SCD were determined using polymerase chain reaction amplification of fragments containing nine polymorphic restriction sites around and within the epsilon-Ggamma-Agamma psibeta-delta-beta-globin gene complex. Most reported haplotypes were found in our population with the Benin haplotype as the most prevalent one. When the patients were divided according to their HbF levels into three groups (Group A: HbF < 5%, Group B: HbF between 5 and 15%, and Group C: HbF > 15%), surprisingly, the highest levels of HbF were associated with the most severe clinical cases. Our findings suggest that fetal hemoglobin levels are important but not the only parameters that affect the severity of the disease. In addition, the high levels of HbF in patients with CAR haplotypes did not seem to ameliorate the severity of symptoms, suggesting that genetic factors other than haplotypes are the major determinants of increased HbF levels in Lebanon. PMID- 12581189 TI - Flow cytometric measurement of reactive oxygen species production by normal and thalassaemic red blood cells. AB - Reactive oxygen species (ROS) contribute to the pathogenesis of several hereditary disorders of red blood cells (RBCs), including thalassaemia. We report here on a modified flow cytometric method for measuring ROS in normal and thalassaemic RBCs. RBCs were incubated with 0.4 mM 2',7'-dichlorofluorescin diacetate (DCFH-DA), then washed and further incubated either with or without 2 mM H2O2. Flow cytometric analysis showed that RBC fluorescence increased with time; it increased faster and reached higher intensity (by 10-30-fold) in H2O2 stimulated RBCs as compared to unstimulated RBCs. In both cases, the antioxidant N-acetyl-l-cysteine reduced fluorescence, confirming previous reports that DCFH fluorescence is mediated by ROS. While the fluorescence of unstimulated RBCs increased with time, probably because of exposure to atmospheric oxygen, in H2O2 stimulated RBCs fluorescence decreased after 30 min. The latter effect is most likely related to H2O2 decomposition by catalase as both sodium azide, an antimetabolite that inhibits catalase and low temperature increased the fluorescence of stimulated RBCs. Washing had a similar effect, suggesting that maintenance of the oxidised DCF requires a constant supply of ROS. We next studied RBCs of beta-thalassaemic patients. The results demonstrated a significantly higher ROS generation by stimulated and unstimulated thalassaemic RBCs compared to their normal counterparts. These results suggest that flow cytometry can be useful for measuring the ROS status of RBCs in various diseases and for studying chemical agents as antioxidants. PMID- 12581190 TI - Low levels of mannose-binding lectin do not affect occurrence of severe infections or duration of fever in acute myeloid leukaemia during remission induction therapy. AB - PURPOSE: To estimate the clinical significance of low serum concentrations of mannose-binding lectin (MBL) in patients with acute myeloid leukaemia (AML) during initial cancer chemotherapy. PATIENTS AND METHODS: 80 consecutive, newly diagnosed, and unselected AML patients (age 18-77 yr) undergoing remission induction chemotherapy. The patients were examined for 28 d. MAIN FINDINGS: Low levels of serum MBL (<1,000 microg/L) were found in 16/80 patients at diagnosis. This frequency is similar to what is found in the general population. In the remaining 64 patients, MBL concentrations were significantly higher than in controls and showed only a slight rise during the period of antineoplastic chemotherapy with its associated infectious complications. Low levels of MBL did not affect overall survival or morbidity in terms of incidence or duration of fever, or occurrence of septicaemia or pneumonia. Long-term survival was likewise independent of MBL concentration. CONCLUSION: MBL levels have no discernible influence on the occurrence or course of infections in AML patients during the initial hospitalisation. The predominant immunodeficiency during this phase is the profound granulocytopenia, which also compromises important effector functions of MBL. The finding in most AML patients of elevated MBL concentrations on admission is most likely because of the role of MBL as an acute phase reactant. PMID- 12581191 TI - Initial exposed phosphatidylserine levels correlate with cellular response to cytotoxic drugs. AB - Phosphatidylserine's (PS) membranal distribution is associated with an expanding variety of biological processes. We studied the relevance of preliminarily exposed membranal PS levels to cellular effects of cytotoxic agents. PBL of normal controls (n = 18) and patients with doxorubicin-treated breast carcinoma (n = 27) or 5'-fluorouracil-treated colorectal cancer (n = 32) were assayed before and after drug infusion. Membranal expression levels of PS, adhesion molecules (CD18, CD11a-c, CD63) and Fas-R of leukocyte subtypes were assessed by flow cytometer. Statistical analysis was implemented. Our results demonstrate external expression of PS on all leukocyte subpopulations despite non-apoptotic light scatter characteristics. Several distinct features were observed of which the more prominent were: leukocyte subtypes each display characteristic PS levels; cancer patients' PBL display higher preliminary PS levels than normal controls in all cell groups; and existence of negative correlations between initial membranal PS levels and drug-induced changes in its expression. Our findings underscore the complex involvement of PS in PBL apoptosis and possibly drug resistance. PMID- 12581192 TI - Diaminofluorene stain detects erythroid differentiation in immature haemopoietic cells treated with EPO, IL-3, SCF, TGFbeta1, MIP-1alpha and IFNgamma. AB - We have combined in vitro clonogenic culture and a highly sensitive stain for haemoglobin to compare the influence of EPO, IL-3, SCF, TGFbeta1, MIP-1alpha and IFNgamma, to directly stimulate cells in the progenitor compartment to develop towards the erythroid lineage. Three cell lines were chosen, as they exist developmentally arrested in the progenitor compartment, yet in a pliant state of maturation. HEL (erythroleukaemia) and K562 (CML-derived) cell lines, may, under appropriate stimuli, develop erythroid characters, whilst the third, U937 (as control cell line), may be stimulated by DMSO to differentiate to myeloid cells. After in vitro semi-solid methylcellulose culture with these cytokines, resulting colonies were stained with 2,7-diaminofluorene (DAF), which sensitively stains haemoglobin blue. Haemoglobin production was low in HEL and K562 cells and absent in U937. Cytokine analysis showed varying levels of influence depending on the starting level of cell line maturation. EPO and TGFbeta1 maximally stimulated haemoglobin production in the HEL and K562 cell lines. This differential cytokine stimulation analysis combined with sensitive DAF haemoglobin detection could be applied in the study of many erythropoiesis-deficient patients or primitive erythropoiesis. PMID- 12581193 TI - A case of multicentric Castleman's disease demonstrating severe eosinophilia and enhanced production of interleukin-5. AB - INTRODUCTION: Castleman's disease (CD), idiopathic lymph-node hyperplasia, is a heterogeneous disease of unknown origin. Although the pathophysiology is yet to be elucidated, interleukin (IL)-6 produced by swollen lymph nodes has been reported to play a crucial role in CD. CASE REPORT: This report presents a case of a 37-yr-old man with CD showing marked elevation of IL-6, hypereosinophilia, and also IL-5 elevation. To date, IL-5 has not been reported to influence CD. However, because in this case the serum concentration of IL-5 produced from swollen lymph nodes paralleled the general symptoms, IL-5, in addition to IL-6, may have played an important role in the disease. DISCUSSION: Although in CD the serum IL-5 level is generally within the normal range, mild eosinophilia is a common complication of the disease. This case indicates that IL-5 can also influence eosinophilia. Taken together, this suggests that there may be a more general association between local production of IL-5 and the pathology of CD. PMID- 12581194 TI - Simultaneous development of lymphoplasmacytic lymphoma and diffuse large B-cell lymphoma--analyses of the clonal relatedness by sequencing CDR3 in immunoglobulin heavy chain genes. AB - A 75-yr-old male simultaneously having lymphoplasmacytic lymphoma (LPL) and diffuse large B-cell lymphoma (DLBCL) is presented. He had manifested with lumbago, high-grade fever, and confusion. Physical examination on admission showed disorientation and severe back pain. There were neither lymphadenopathy nor hepatosplenomegaly. Routine laboratory tests showed moderate pancytopenia, hypercalcemia (serum calcium, 15.9 mg/dL), IgM lambda-type monoclonal gammopathy (IgG, 405 mg/dL; IgA, 42 mg/dL; and IgM, 2023 mg/dL), and lambda-type Bence-Jones protein in the urine (0.8 g/d). Bone marrow biopsy showed the clusters of surface lambda-positive small-sized mature-appearing lymphoplasmacytoid cells. Bone survey and computed tomographic scan showed multiple osteolytic lesions and a tumor involving the third lumbar spine (L3). An open biopsy of the L3 tumor showed diffuse proliferation of CD20- and lambda-positive large cells. We thus diagnosed the patient as simultaneously having LPL and DLBCL. Although the combination chemotherapy was at least partially effective, he died of bacteremia and organ failure after three courses of chemotherapy. To clarify the clonal relatedness between LPL and DLBCL, we analyzed the sequences of the complementarity-determining region 3 in immunoglobulin heavy-chain genes. The data showed that LPL and DLBCL in the present patient originated from two independent clones. PMID- 12581195 TI - Rebound hepatosplenomegaly in type 1 Gaucher disease. AB - A 19-year-old male patient with type 1 Gaucher disease was put on regular biweekly infusions of alglucerase. After 1 yr of treatment, hepatic and splenic volumes decreased from 38 and 45 mL/kg to 31 and 34 mL/kg, respectively. In addition, hemoglobin concentration, platelet count and white cell count increased, acid phosphatase level decreased, and the patient gained weight and energy. Despite improvement, the patient refused enzyme replacement therapy (ERT) because of muscle rigidity, chest pain, trembling and anxiety, which he attributed to enzyme substitution. Two and 4.5 year after cessation of therapy, hepatic and splenic volumes increased to 36 and 53 mL/kg and to 53 and 110 mL/kg, respectively. The patient developed non-tractable hematuria because of compression and dislocation of the left kidney by the enlarged spleen, which necessitated splenectomy. This report suggests that cessation of ERT in Gaucher disease may result in severe and complicated rebound visceromegaly. PMID- 12581196 TI - Emerging strategies in mass-spectrometry based proteomics. PMID- 12581197 TI - Affinity purification-mass spectrometry. Powerful tools for the characterization of protein complexes. AB - Multi-protein complexes are emerging as important entities of biological activity inside cells that serve to create functional diversity by contextual combination of gene products and, at the same time, organize the large number of different proteins into functional units. Many a time, when studying protein complexes rather than individual proteins, the biological insight gained has been fundamental, particularly in cases in which proteins with no previous functional annotation could be placed into a functional context derived from their 'molecular environment'. In this minireview, we summarize the current state of the art for the retrieval of multiprotein complexes by affinity purification and their analysis by mass spectrometry. The advances in technology made over the past few years now enable the study of protein complexes on a proteomic scale and it can be anticipated that the knowledge gathered from such projects will fuel drug target discovery and validation pipelines and that the technology is also going to prove valuable in the emerging field of systems biology. PMID- 12581198 TI - Deciphering metabolic networks. AB - All higher organisms divide major biochemical steps into different cellular compartments and often use tissue-specific division of metabolism for the same purpose. Such spatial resolution is accompanied with temporal changes of metabolite synthesis in response to environmental stimuli or developmental needs. Although analyses of primary and secondary gene products, i.e. transcripts, proteins, and metabolites, regularly do not cope with this spatial and temporal resolution, these gene products are often observed to be highly coregulated forming complex networks. Methods to study such networks are reviewed with respect to data acquisition, network statistics, and biochemical interpretation. PMID- 12581199 TI - Proteome analysis at the level of subcellular structures. AB - The targeting of proteins to particular subcellular sites is an important principle of the functional organization of cells at the molecular level. In turn, knowledge about the subcellular localization of a protein is a characteristic that may provide a hint as to the function of the protein. The combination of classic biochemical fractionation techniques for the enrichment of particular subcellular structures with the large-scale identification of proteins by mass spectrometry and bioinformatics provides a powerful strategy that interfaces cell biology and proteomics, and thus is termed 'subcellular proteomics'. In addition to its exceptional power for the identification of previously unknown gene products, the analysis of proteins at the subcellular level is the basis for monitoring important aspects of dynamic changes in the proteome such as protein transloction. This review summarizes data from recent subcellular proteomics studies with an emphasis on the type of data that can retrieved from such studies depending on the design of the analytical strategy. PMID- 12581200 TI - A simple protocol to study blue copper proteins by NMR. AB - In the case of oxidized plastocyanin from Synechocystis sp. PCC6803, an NMR approach based on classical two and three dimensional experiments for sequential assignment leaves unobserved 14 out of 98 amino acids. A protocol which simply makes use of tailored versions of 2D HSQC and 3D CBCA(CO)NH and CBCANH leads to the identification of nine of the above 14 residues. The proposed protocol differs from previous approaches in that it does not involve the use of unconventional experiments designed specifically for paramagnetic systems, and does not exploit the occurrence of a corresponding diamagnetic species in chemical exchange with the blue copper form. This protocol is expected to extend the popularity of NMR in the structural studies of copper (II) proteins, allowing researchers to increase the amount of information available via NMR on the neighborhood of a paramagnetic center without requiring a specific expertise in the field. The resulting 3D spectra are standard spectra that can be handled by any standard software for protein NMR data analysis. PMID- 12581201 TI - Structural diversity in lipopolysaccharide expression in nontypeable Haemophilus influenzae. Identification of L-glycerol-D-manno-heptose in the outer-core region in three clinical isolates. AB - Structural elucidation of the lipopolysaccharide (LPS) from three nontypeable Haemophilus influenzae clinical isolates, 1209, 1207 and 1233 was achieved using NMR spectroscopy and ESI-MS on O-deacylated LPS and core oligosaccharide (OS) material as well as ESI-MS(n) on permethylated dephosphorylated OS. It was found that the organisms expressed a tremendous heterogeneous glycoform mixture resulting from the variable length of the OS chains attached to the common structural element of H. influenzae, L-alpha-D-Hepp-(1-->2)-[PEtn-->6]-L-alpha-D Hepp-(1-->3)-[beta-D-Glcp-(1-->4)]-L-alpha-D-Hepp-(1-->5)-[PPEtn-->4]-alpha-Kdop (2-->6)-Lipid A. Notably, the O-6 position of the beta-D-Glcp residue could either be occupied by PCho or L-glycero-D-manno-heptose (L,D-Hep), which is a location for L,D-Hep that has not been seen previously in H. influenzae LPS. The outer-core L,D-Hep residue was further chain elongated at the O-6 position by the structural element beta-D-GalpNAc-(1-->3)-alpha-D-Galp-(1-->4)-beta-D-Galp, or sequentially truncated versions thereof. The distal heptose residue in the inner core was found to be chain elongated at O-2 by the globotetraose unit, beta-D GalpNAc-(1-->3)-alpha-D-Galp-(1-->4)-beta-D-Galp-(1-->4)-beta-D-Glcp, or sequentially truncated versions thereof. Investigation of LPS from an lpsA mutant of isolate 1233 and a lic1 mutant of isolate 1209 was also performed, which aside from confirming the functions of the gene products, simplified elucidation of the OS extending from the proximal heptose (the lpsA mutant), and showed that the organism exclusively expresses LPS glycoforms comprising the outer-core l,d-Hep residue when PCho is not expressed (the lic1 mutant). PMID- 12581202 TI - Cytotoxic activity of nucleoside diphosphate kinase secreted from Mycobacterium tuberculosis. AB - Pathogenicity of Mycobacterium tuberculosis is closely related to its ability to survive and replicate in the hostile environment of macrophages. For some pathogenic bacteria, secretion of ATP-utilizing enzymes into the extracellular environment aids in pathogen survival via P2Z receptor-mediated, ATP-induced death of infected macrophages. A component of these enzymes is nucleoside diphosphate kinase (Ndk). The ndk gene was cloned from M. tuberculosis H37Rv and expressed in Escherichia coli. Ndk was secreted into the culture medium by M. tuberculosis, as determined by enzymatic activity and Western blotting. Purified Ndk enhanced ATP-induced macrophage cell death, as assayed by the release of [14C]adenine. A catalytic mutant of Ndk failed to enhance ATP-induced macrophage cell death, and periodate-oxidized ATP (oATP), an irreversible inhibitor of P2Z receptor, blocked ATP/Ndk-induced cell death. Purified Ndk was also found to be autophosphorylated with broad specificity for all nucleotides. Conversion of His117-->Gln, which is part of the nucleotide-binding site, abolished autophosphorylation. Purified Ndk also showed GTPase activity. Collectively, these results indicate that secreted Ndk of M. tuberculosis acts as a cytotoxic factor for macrophages, which may help in dissemination of the bacilli and evasion of the immune system. PMID- 12581203 TI - Relation between domain evolution, specificity, and taxonomy of the alpha-amylase family members containing a C-terminal starch-binding domain. AB - The alpha-amylase family (glycoside hydrolase family 13; GH 13) contains enzymes with approximately 30 specificities. Six types of enzyme from the family can possess a C-terminal starch-binding domain (SBD): alpha-amylase, maltotetraohydrolase, maltopentaohydrolase, maltogenic alpha-amylase, acarviose transferase, and cyclodextrin glucanotransferase (CGTase). Such enzymes are multidomain proteins and those that contain an SBD consist of four or five domains, the former enzymes being mainly hydrolases and the latter mainly transglycosidases. The individual domains are labelled A [the catalytic (beta/alpha)8-barrel], B, C, D and E (SBD), but D is lacking from the four-domain enzymes. Evolutionary trees were constructed for domains A, B, C and E and compared with the 'complete-sequence tree'. The trees for domains A and B and the complete-sequence tree were very similar and contain two main groups of enzymes, an amylase group and a CGTase group. The tree for domain C changed substantially, the separation between the amylase and CGTase groups being shortened, and a new border line being suggested to include the Klebsiella and Nostoc CGTases (both four-domain proteins) with the four-domain amylases. In the 'SBD tree' the border between hydrolases (mainly alpha-amylases) and transglycosidases (principally CGTases) was not readily defined, because maltogenic alpha-amylase, acarviose transferase, and the archaeal CGTase clustered together at a distance from the main CGTase cluster. Moreover the four-domain CGTases were rooted in the amylase group, reflecting sequence relationships for the SBD. It appears that with respect to the SBD, evolution in GH 13 shows a transition in the segment of the proteins C-terminal to the catalytic (beta/alpha)8-barrel(domain A). PMID- 12581204 TI - Lep d 2 polymorphisms in wild and cultured Lepidoglyphus destructor mites. AB - We have previously cloned, expressed and characterized two variants of the major allergen Lep d 2 from cultured Lepidoglyphus destructor mites. These variants, Lep d 2.0101 and Lep d 2.0201, differ at 13 amino acid positions. In this study we investigated Lep d 2 sequence diversity between wild and cultured mites. PCR, Southern blot and DNA sequence analysis revealed the presence of two different Lep d 2 genes, one with and one without an intron. In addition, two new variants of Lep d 2, Lep d 2.0102 and Lep d 2.0202, were found at different frequencies in wild and cultured mites. When we expressed the Lep d 2 variants and compared their IgE binding properties by ELISA inhibition, we found that Lep d 2.0102 was a more potent inhibitor than Lep d 2.0101, and to a lesser extent Lep d 2.0202 was more potent than Lep d 2.0201. Long-term cultures of peripheral blood mononuclear cells were used to assess the ability of the expressed Lep d 2 variants to induce cytokine release. Although cells from different individuals released different amounts of interferon-gamma and interleukin-5, no consistent cytokine release pattern could be linked to any specific Lep d 2 variant. In conclusion, we show that both cultured and wild Lepidoglyphus destructor mites contain the same pattern of polymorphism. Furthermore, this Lep d 2 sequence diversity seems not to have any significant impact on the allergens IgE binding or its ability to induce T cell cytokine release. PMID- 12581205 TI - Co-incorporation of A beta 40 and A beta 42 to form mixed pre-fibrillar aggregates. AB - Senile plaques, the invariable hallmark and likely proximal cause of Alzheimer's disease (AD), are structured depositions of the 40- and 42-residue forms of the A beta peptide. Conversely, diffuse plaques, which are not associated with neurodegeneration, consist mainly of unstructured A beta 42. We have investigated the interaction between A beta 40 and A beta 42 through an assay, which involves labeling both variants with an environment-sensitive fluorophore. We have monitored association of A beta without fibrillar seeds, which allows investigation of molecular species preceding fibrils. Immediately upon mixture, A beta 40 and A beta 42 associate into mixed aggregates, in which the peptides are unstructured and relatively accessible to water. When left to incubate for an extended period, larger, more tightly packed aggregates, which show secondary structure, replace the small, unstructured aggregates formed earlier. Our results show that in vitro the two A beta variants coassemble early in the fibrillogenesis pathway. The ease of formation for mixed and homogeneous aggregates is similar. A change in the local A beta variant ratio can therefore have a significant impact on A beta aggregation; indeed such a change has been reported in some types of familial AD. PMID- 12581206 TI - Functionally distinct dopamine and octopamine transporters in the CNS of the cabbage looper moth. AB - A cDNA was cloned from the cabbage looper Trichoplusia ni based on similarity to other cloned dopamine transporters (DATs). The total nucleotide sequence is 3.8 kb in length and contains an open reading frame for a protein of 612 amino acids. The predicted moth DAT protein (TrnDAT) has greatest amino acid sequence identity with Drosophila melanogasterDAT (73%) and Caenorhabditis elegansDAT (51%). TrnDAT shares only 45% amino acid sequence identity with an octopamine transporter (TrnOAT) cloned recently from this moth. The functional properties of TrnDAT and TrnOAT were compared through transient heterologous expression in Sf9 cells. Both transporters have similar transport affinities for DA (Km 2.43 and 2.16 micro m, respectively). However, the competitive substrates octopamine and tyramine are more potent blockers of [3H]dopamine (DA) uptake by TrnOAT than by TrnDAT. D Amphetamine is a strong inhibitor and l-norepinephrine a weak inhibitor of both transporters. TrnDAT-mediated DA uptake is approximately 100-fold more sensitive to selective blockers of vertebrate transporters of dopamine and norepinephrine, such as nisoxetine, nomifensine and dibenzazepine antidepressants, than TrnOAT mediated DA uptake. TrnOAT is 10-fold less sensitive to cocaine than TrnDAT. None of the 15 monoamine uptake blockers tested was TrnOAT-selective. In situ hybridization shows that TrnDAT and TrnOAT transcripts are expressed by different sets of neurons in caterpillar brain and ventral nerve cord. These results show that the caterpillar CNS contains both a phenolamine transporter and a catecholamine transporter whereas in the three invertebrates whose genomes have been completely sequenced only a dopamine-selective transporter is found. PMID- 12581207 TI - Purification and characterization of three isoforms of chrysophsin, a novel antimicrobial peptide in the gills of the red sea bream, Chrysophrys major. AB - We report here the isolation of three isoforms of a novel C-terminally amidated peptide from the gills of red sea bream, Chrysophrys (Pagrus) major. Peptide sequences were determined by a combination of Edman degradation, MS and HPLC analysis of native and synthetic peptides. Three peptides, named chrysophsin-1, chrysophsin-2, and chrysophsin-3, consist of 25, 25, and 20 amino acids, respectively, and are highly cationic, containing an unusual C-terminal RRRH sequence. The alpha-helical structures of the three chrysophsin peptides were predicted from their secondary structures and were confirmed by CD spectroscopy. The synthetic peptides displayed broad-spectrum bactericidal activity against Gram-negative and Gram-positive bacteria including Escherichia coli, Bacillus subtilis, and fish and crustacean pathogens. The three peptides were also hemolytic. Immunohistochemical analysis showed that chrysophsins were localized in certain epithelial cells lining the surface of secondary lamellae and eosinophilic granule cell-like cells at the base of the secondary lamellae in red sea bream gills. Their broad ranging bactericidal activities, combined with their localization in certain cells and eosinophilic granule cell-like cells in the gills, suggest that chrysophsins play a significant role in the innate defense system of red sea bream gills. PMID- 12581208 TI - Expression and characterization of cyanobacterium heme oxygenase, a key enzyme in the phycobilin synthesis. Properties of the heme complex of recombinant active enzyme. AB - An efficient bacterial expression system of cyanobacterium Synechocystis sp. PCC 6803 heme oxygenase gene, ho-1, has been constructed, using a synthetic gene. A soluble protein was expressed at high levels and was highly purified, for the first time. The protein binds equimolar free hemin to catabolize the bound hemin to ferric-biliverdin IX alpha in the presence of oxygen and reducing equivalents, showing the heme oxygenase activity. During the reaction, verdoheme intermediate is formed with the evolution of carbon monoxide. Though both ascorbate and NADPH cytochrome P450 reductase serve as an electron donor, the heme catabolism assisted by ascorbate is considerably slow and the reaction with NADPH-cytochrome P450 reductase is greatly retarded after the oxy-heme complex formation. The optical absorption spectra of the heme-enzyme complexes are similar to those of the known heme oxygenase complexes but have some distinct features, exhibiting the Soret band slightly blue-shifted and relatively strong CT bands of the high spin component in the ferric form spectrum. The heme-enzyme complex shows the acid-base transition, where two alkaline species are generated. EPR of the nitrosyl heme complex has established the nitrogenous proximal ligand, presumably histidine 17 and the obtained EPR parameters are discriminated from those of the rat heme oxygenase-1 complex. The spectroscopic characters as well as the catabolic activities strongly suggest that, in spite of very high conservation of the primary structure, the heme pocket structure of Synechocystis heme oxygenase isoform-1 is different from that of rat heme oxygenase isoform-1, rather resembling that of bacterial heme oxygenase, H mu O. PMID- 12581209 TI - Beta 3-adrenoceptor knockout in C57BL/6J mice depresses the occurrence of brown adipocytes in white fat. AB - White and brown adipocytes are usually located in distinct depots; however, in response to cold, brown adipocytes appear in white fat. This response is mediated by beta-adrenoceptors but there is a controversy about the subtype(s) involved. In the present study, we exposed to cold beta 3-adrenoceptor knockout mice (beta 3KO) on a C57BL/6J genetic background and measured in white adipose tissue the density of multilocular cells and the expression of the brown adipocyte marker uncoupling protein-1 (UCP1). In brown fat of beta 3KO mice, UCP1 expression levels were normal at 24 degrees C as well as after a 10-day cold exposure. Strikingly, under both conditions, in the white fat of beta 3KO mice the levels of UCP1 mRNA and protein as well as the density of multilocular cells were decreased. These results indicate that beta 3-adrenoceptors play a major role in the appearance of brown adipocytes in white fat and suggest that the brown adipocytes present in white fat differ from those in brown fat. PMID- 12581210 TI - Complex alternative splicing of the hKLK3 gene coding for the tumor marker PSA (prostate-specific-antigen). AB - PSA (prostate-specific antigen), the most useful serum marker for prostate cancer, is encoded by the hKLK3 gene and is present in the serum as a mixture of several molecular species. This work was performed to identify the hKLK3 transcripts in order to determine how many proteins resembling PSA are synthesized from the hKLK3 gene and secreted in blood. Combined Northern blotting, molecular cloning and database searching showed that the hKLK3 gene produces at least 15 transcripts ranging in size from 0.7 to 6.1 kb. Polysomal distribution analysis revealed that the transcripts shorter than 3.1 kb are efficiently translated in prostate cell line. A total of 12 hKLK3 transcripts have been completely or partially cloned. They result from alternative splicing or/and alternative polyadenylation involving complex regulation. They code for eight proteins: PSA, a truncated form of PSA (PSA-Tr), five PSA variants (PSA RPs) and one protein (PSA-LM) unrelated to PSA. Using a specific antibody, we detected the PSA-RP2 variant in prostate tissue. All the variants share the same signal peptide and could contribute to the diversity of hKLK3 proteins in prostate fluid and blood. PMID- 12581211 TI - Structure, mRNA expression and linkage mapping of the brain-type fatty acid binding protein gene (FABP7) from zebrafish (Danio rerio). AB - The brain fatty acid-binding protein (B-FABP) is involved in brain development and adult neurogenesis. We have determined the sequence of the gene encoding the B-FABP in zebrafish. The zebrafish B-FABP gene spans 2370 bp and contains four exons interrupted by three introns. The coding sequence of zebrafish B-FABP gene is identical to its cDNA sequence and the coding capacity of each exon is the same as that for the human and mouse B-FABP genes. A 1249 bp sequence 5' upstream of exon 1 of the zebrafish B-FABP gene was cloned and sequenced. Several brain development/growth-associated transcription factor binding elements, including POU-domain binding elements and the proposed lipogenic-associated transcription factor NF-Y elements, were found within the 5' region of the B-FABP gene. RT-PCR analysis using mRNA extracted from different tissues of adult zebrafish demonstrated that the zebrafish B-FABP mRNA was predominant in brain with lower levels in liver, testis and intestine, but not in ovary, skin, heart, kidney and muscle. Quantitative RT-PCR revealed a similar tissue-specific distribution for zebrafish B-FABP mRNA except that very low levels of B-FABP mRNA, normalized to beta-actin mRNA, were detected in the heart and muscle RNA, but not in liver RNA. Zebrafish B-FABP mRNA was detected by RT-PCR in embryos beyond 12 h postfertilization, suggesting a correlation of zebrafish B-FABP mRNA expression with early brain development. Radiation hybrid mapping assigned the zebrafish B FABP gene to linkage group 17. Conserved syntenies of the zebrafish B-FABP gene and the human and mouse orthologous B-FABP genes were observed by comparative genomic analysis. PMID- 12581212 TI - Insights into the design of a hybrid system between Anabaena ferredoxin-NADP+ reductase and bovine adrenodoxin. AB - The opportunity to design enzymatic systems is becoming more feasible due to detailed knowledge of the structure of many proteins. As a first step, investigations have aimed to redesign already existing systems, so that they can perform a function different from the one for which they were synthesized. We have investigated the interaction of electron transfer proteins from different systems in order to check the possibility of heterologous reconstitution among members of different chains. Here, it is shown that ferredoxin-NADP+ reductase from Anabaena and adrenodoxin from bovine adrenal glands are able to form optimal complexes for thermodynamically favoured electron transfer reactions. Thus, electron transfer from ferredoxin-NADP+ reductase to adrenodoxin seems to proceed through the formation of at least two different complexes, whereas electron transfer from adrenodoxin to ferredoxin-NADP+ reductase does not take place due because it is a thermodynamically nonfavoured process. Moreover, by using a truncated adrenodoxin form (with decreased reduction potential as compared with the wild-type) ferredoxin-NADP+ reductase is reduced. Finally, these reactions have also been studied using several ferredoxin-NADP+ reductase mutants at positions crucial for interaction with its physiological partner, ferredoxin. The effects observed in their reactions with adrenodoxin do not correlate with those reported for their reactions with ferredoxin. In summary, our data indicate that although electron transfer can be achieved in this hybrid system, the electron transfer processes observed are much slower than within the physiological partners, pointing to a low specificity in the interaction surfaces of the proteins in the hybrid complexes. PMID- 12581213 TI - Characterization of acetyl-CoA/propionyl-CoA carboxylase in Metallosphaera sedula. Carboxylating enzyme in the 3-hydroxypropionate cycle for autotrophic carbon fixation. AB - Autotrophic Archaea of the family Sulfolobaceae (Crenarchaeota) use a modified 3 hydroxypropionate cycle for carbon dioxide assimilation. In this cycle the ATP dependent carboxylations of acetyl-CoA and propionyl-CoA to malonyl-CoA and methylmalonyl-CoA, respectively, represent the key CO2 fixation reactions. These reactions were studied in the thermophilic and acidophilic Metallosphaera sedula and are shown to be catalyzed by one single large enzyme, which acts equally well on acetyl-CoA and propionyl-CoA. The carboxylase was purified and characterized and the genes were cloned and sequenced. In contrast to the carboxylase of most other organisms, acetyl-CoA/propionyl-CoA carboxylase from M. sedula is active at 75 degrees C and is isolated as a stabile functional protein complex of 560 +/- 50 kDa. The enzyme consists of two large subunits of 57 kDa each representing biotin carboxylase (alpha) and carboxytransferase (gamma), respectively, and a small 18.6 kDa biotin carrier protein (beta). These subunits probably form an (alpha beta gamma)4 holoenzyme. It has a catalytic number of 28 s-1 at 65 degrees C and at the optimal pH of 7.5. The apparent Km values were 0.06 mm for acetyl CoA, 0.07 mm for propionyl-CoA, 0.04 mm for ATP and 0.3 mm for bicarbonate. Acetyl-CoA/propionyl-CoA carboxylase is considered the main CO2 fixation enzyme of autotrophic members of Sulfolobaceae and the sequenced genomes of these Archaea contain the respective genes. Due to its stability the archaeal carboxylase may prove an ideal subject for further structural studies. PMID- 12581214 TI - Unusual fluorescence of W168 in Plasmodium falciparum triosephosphate isomerase, probed by single-tryptophan mutants. AB - Plasmodium falciparum triosephosphate isomerase (PfTIM) contains two tryptophan residues, W11 and W168. One is positioned in the interior of the protein, and the other is located on the active-site loop 6. Two single-tryptophan mutants, W11F and W168F, were constructed to evaluate the contributions of each chromophore to the fluorescence of the wild-type (wt) protein and to probe the utility of the residues as spectroscopic reporters. A comparative analysis of the fluorescence spectra of PfTIMwt and the two mutant proteins revealed that W168 possesses an unusual, blue-shifted emission (321 nm) and exhibits significant red-edge excitation shift of fluorescence. In contrast, W11 emits at 332 nm, displays no excitation dependence of fluorescence, and behaves like a normal buried chromophore. W168 has a much shorter mean lifetime (2.7 ns) than W11 (4.6 ns). The anomalous fluorescence properties of W168 are abolished on unfolding of the protein in guanidinium chloride (GdmCl) or at low pH. Analysis of the tryptophan environment using a 1.1-A crystal structure established that W168 is rigidly held by a complex network of polar interactions including a strong hydrogen bond from Y164 to the indole NH group. The environment is almost completely polar, suggesting that electrostatic effects determine the unusually low emission wavelength of W168. To our knowledge this is a unique observation of a blue shifted emission from a tryptophan in a polar environment in the protein. The wild-type and mutant proteins show similar levels of enzymatic activity and secondary and tertiary structure. However, the W11F mutation appreciably destabilizes the protein to unfolding by urea and GdmCl. The fluorescence of W168 is shown to be extremely sensitive to binding of the inhibitor, 2-phosphoglycolic acid. PMID- 12581215 TI - Mapping of chorismate mutase and prephenate dehydrogenase domains in the Escherichia coli T-protein. AB - The Escherichia coli bifunctional T-protein transforms chorismic acid to p hydroxyphenylpyruvic acid in the l-tyrosine biosynthetic pathway. The 373 amino acid T-protein is a homodimer that exhibits chorismate mutase (CM) and prephenate dehydrogenase (PDH) activities, both of which are feedback-inhibited by tyrosine. Fifteen genes coding for the T-protein and various fragments thereof were constructed and successfully expressed in order to characterize the CM, PDH and regulatory domains. Residues 1-88 constituted a functional CM domain, which was also dimeric. Both the PDH and the feedback-inhibition activities were localized in residues 94-373, but could not be separated into discrete domains. The activities of cloned CM and PDH domains were comparatively low, suggesting some cooperative interactions in the native state. Activity data further indicate that the PDH domain, in which NAD, prephenate and tyrosine binding sites were present, was more unstable than the CM domain. PMID- 12581216 TI - A comparative analysis of the time-dependent antiproliferative effects of daunorubicin and WP631. AB - Jurkat T lymphocytes were treated with daunorubicin and WP631, a daunorubicin based DNA binding agent, in experiments aimed to analyze cellular uptake of these drugs and their effect on cell viability. WP631 was taken up more slowly than daunorubicin, but laser confocal microscopy and spectrofluorometric quantification showed that the drug accumulated in the cells. Despite the slow uptake rate, the antiproliferative capacity of WP631 (measured as IC50 after a 72 h continuous treatment) was greater than that of daunorubicin. The propensities of daunorubicin and WP631 to promote apoptosis were compared. Our results indicate that the major effect of WP631 was a G2/M arrest followed, after about 72 h of treatment, by polyploidy and mitotic (reproductive) death. In contrast, daunorubicin induced a rapid response with classic features of apoptosis. PMID- 12581217 TI - Purification and cDNA cloning of a cellulase from abalone Haliotis discus hannai. AB - A cellulase [endo-beta-1,4-D-glucanase (EC 3.2.1.4)] was isolated from the hepatopancreas of abalone Haliotis discus hannai by successive chromatographies on TOYOPEARL CM-650M, hydroxyapatite and Sephacryl S-200 HR. The molecular mass of the cellulase was estimated to be 66 000 Da by SDS/PAGE, thus the enzyme was named HdEG66. The hydrolytic activity of HdEG66 toward carboxymethylcellulose showed optimal temperature and pH at 38 degrees C and 6.3, respectively. cDNAs encoding HdEG66 were amplified by the polymerase chain reaction from an abalone hepatopancreas cDNA library with primers synthesized on the basis of partial amino-acid sequences of HdEG66. By overlapping the nucleotide sequences of the cDNAs, a sequence of 1898 bp in total was determined. The coding region of 1785 bp located at nucleotide position 56-1840 gave an amino-acid sequence of 594 residues including the initiation methionine. The N-terminal region of 14 residues in the deduced sequence was regarded as the signal peptide as it was absent in HdEG66 protein and showed high similarity to the consensus sequence for signal peptides of eukaryote secretory proteins. Thus, matured HdEG66 was thought to consist of 579 residues. The C-terminal region of 453 residues in HdEG66, i.e. approximately the C-terminal three quarters of the protein, showed 42-44% identity to the catalytic domains of glycoside hydrolase family 9 (GHF9) cellulases from arthropods and Thermomonospora fusca. While the N-terminal first quarter of HdEG66 showed 27% identity to the carbohydrate-binding module (CBM) of a Cellulomonas fimi cellulase, CenA. Thus, the HdEG66 was regarded as the GHF9 cellulase possessing a family II CBM in the N-terminal region. By genomic PCR using specific primers to the 3'-terminal coding sequences of HdEG66-cDNA, a DNA of 2186 bp including three introns was amplified. This strongly suggests that the origin of HdEG66 is not from symbiotic bacteria but abalone itself. PMID- 12581219 TI - Of cabbages and kings: some considerations on classifications, diagnostic schemes, semiology, and concepts. PMID- 12581224 TI - Effects of topiramate in two models of genetically determined generalized epilepsy, the GAERS and the Audiogenic Wistar AS. AB - PURPOSE: The antiepileptic effects of topiramate (TPM) were assessed in two models of genetically determined generalized epilepsy. The model of nonconvulsive epilepsy used was a model of absence seizures, the GAERS (Genetic Absence Epilepsy Rat from Strasbourg); and the model of convulsive seizures was an audiogenic rat model, the Wistar Audiogenic Sensitive (AS) rat. METHODS: GAERS were equipped with four cortical electrodes over the frontoparietal cortex, and the duration of spike-and-wave discharges (SWDs) on the EEG was recorded for periods of 20 to 120 or 300 min. In Wistar AS, the occurrence of, latency to, and duration of one or two wild running episodes and tonic seizures were recorded. RESULTS: In the 16 GAERS studied, TPM (10, 30, and 60 mg/kg) dose-dependently reduced the expression of SWD that almost totally disappeared at the two highest doses between 40 and 120 min. SWD duration returned to control levels by 180 and 280 min after the injection of 30 and 60 mg/kg TPM, respectively. In Wistar AS, 10 mg/kg TPM induced the occurrence of a second running episode not present in control rats, indicative of a decrease in sensitivity of the rats to the stimulus and increased by 330% the latency to the tonic seizure that still occurred in the eight rats studied. At 30 and 60 mg/kg, the latency to wild running increased by 140%; the second running episode was suppressed in six and seven rats, respectively, whereas the tonic seizure occurred only in one of the eight rats studied at these two doses. CONCLUSIONS: These results support the broad spectrum of antiepileptic activity of TPM, confirming its efficacy in primary generalized seizures of both tonic-clonic and of the absence type. PMID- 12581225 TI - Ih blockers have a potential of antiepileptic effects. AB - PURPOSE: The h current (Ih) is an inwardly mixed cationic conductance activated by membrane hyperpolarization and distributed predominantly in the apical dendrites of hippocampal pyramidal neurons. To verify a hypothesis that an anomalous hyperpolarization generates an abnormal excitation by way of Ih channels, we examined the effects of Ih blockers (CsCl and ZD7288) on electrically induced paroxysmal discharges (PADs). METHODS: Fifty-three adult male rabbits were used. We measured the PAD threshold elicited by stimulation to the apical dendritic layer of the hippocampal CA1 region before and after injecting 50 microl of each Ih blocker or saline extracellularly into the same region. RESULTS: In Ih blocker injection groups (n = 26), we obtained a significant increase in PAD threshold (1 mM CsCl: 163%, p < 0.01; 10 mM CsCl: 265%, p < 0.01; 100 mM CsCl: 199%, p < 0.01; 100 microM ZD7288: 192%, p < 0.05; 1 mM ZD7288: 246%, p < 0.05). Conversely, we did not obtain the increase in PAD threshold in a saline injection group (n = 10, 107%). The magnitude as well as duration of the effect had a tendency to depend on concentration of Ih blockers, although a saturated or declining tendency was observed with the 100 mM CsCl injection. CONCLUSIONS: We concluded that Ih channels might contribute to hippocampal epileptiform discharges in vivo. Our hypothesis for epileptogenesis demonstrated in the present experiment offers an idea to develop a new type of antiepileptic drug based on Ih blockers for the treatment of epileptic disorders refractory to current medications. PMID- 12581226 TI - Sexual dysfunction and sudden death in epileptic male EL mice: inheritance and prevention with the ketogenic diet. AB - PURPOSE: The EL mouse is an animal model for multifactorial idiopathic epilepsy. Although EL mice have been studied extensively for >45 years, the etiology of male sudden death and its relation to seizures have not been defined. Here we investigated the cause of EL male sudden death and its relation to epilepsy. METHODS: For histopathologic analysis, the terminally ill EL mice (n = 15) were killed, and the tissues were fixed. Blood chemical composition was compared between the terminally ill EL (n = 9) and the healthy age-matched EL (n = 17) and DDY (n = 11) males. To determine the effect of the ketogenic diet (KD) on sudden male death, young male EL mice (P30) were randomly separated into two groups that were fed ad libitum with either Agway lab chow (control n = 38) or with the KD (treated, n = 39) for 5 months. The genetic predisposition to sudden death was analyzed in the backcross generation (n = 106) of a cross between EL and the nonepileptic ABP strains. RESULTS: Sudden death coincided with the onset of seizures (70-80 days) and affected 94% of male EL mice by age 300 days. Urethral plugs were observed histologically in 13 of 15 longitudinally sectioned penises. Concentrations of blood urea nitrogen, creatinine, phosphorus, and calcium in the terminally ill mice were significantly elevated when compared with those of healthy animals. None of the mice treated with the KD experienced sudden death, whereas 15 (39%) of the untreated control mice died by age 5 months. The sudden death in male EL mice was inherited as an autosomal recessive sex-limited lethal trait. CONCLUSIONS: The cause of sudden death in male EL mice arises from abnormal ejaculation, which produces a urethral plug with consequent urinary retention and acute severe uremia. The coincident onset of seizures and sudden death in EL males suggests that a sexual dysfunction is associated with epilepsy in this model. PMID- 12581227 TI - Exploration of a putative susceptibility locus for idiopathic generalized epilepsy on chromosome 8p12. AB - PURPOSE: A recent genome-wide scan revealed a major susceptibility locus for idiopathic generalized epilepsies (IGEs) in the chromosomal region 8p12 in 32 IGE families without members with juvenile myoclonic epilepsy (JME). This study explored the presence of an IGE locus in the chromosomal region 8p12. METHODS: Our study included 176 multiplex families of probands with common IGE syndromes. Parametric and nonparametric multipoint linkage analyses were carried out between the IGE trait and six microsatellite polymorphisms encompassing the putative susceptibility locus. To explore the associated phenotype-genotype relation, two distinct subgroups of families were selected by the presence (n = 64) or absence (n = 112) of a family member with JME. To adjust the phenotypic spectrum toward adolescent-onset IGEs, a third subgroup of 28 families without JME was chosen through an IGE proband with seizure onset at age 10-20 years. RESULTS: Parametric and nonparametric multipoint linkage analyses provided no evidence for linkage between IGE and markers encompassing the putative IGE locus in the chromosomal region 8p12. Furthermore, we found no hint of linkage along the candidate region in any of the three family subgroups. CONCLUSIONS: We failed to provide evidence for a major IGE locus in the chromosomal region 8p12. On the contrary, these parametric linkage results provide strong evidence against linkage across the candidate region under a broad range of genetic models. If there is a susceptibility locus for IGE in the chromosomal region 8p12, then the size of the effect or the proportion of linked families is too small to detect linkage in the investigated family sample. PMID- 12581228 TI - Interlaboratory variability in the quantification of new generation antiepileptic drugs based on external quality assessment data. AB - PURPOSE: To assess interlaboratory variability in the determination of serum levels of new antiepileptic drugs (AEDs). METHODS: Lyophilised serum samples containing clinically relevant concentrations of felbamate (FBM), gabapentin (GBP), lamotrigine (LTG), the monohydroxy derivative of oxcarbazepine (OCBZ; MHD), tiagabine (TGB), topiramate (TPM), and vigabatrin (VGB) were distributed monthly among 70 laboratories participating in the international Heathcontrol External Quality Assessment Scheme (EQAS). Assay results returned over a 15-month period were evaluated for precision and accuracy. RESULTS: The most frequently measured compound was LTG (65), followed by MHD (39), GBP (19), TPM (18), VGB (15), FBM (16), and TGB (8). High-performance liquid chromatography was the most commonly used assay technique for all drugs except for TPM, for which two thirds of laboratories used a commercial immunoassay. For all assay methods combined, precision was <11% for MHD, FBM, TPM, and LTG, close to 15% for GBP and VGB, and as high as 54% for TGB (p < 0.001). Mean accuracy values were <10% for all drugs other than TGB, for which measured values were on average 13.9% higher than spiked values, with a high variability around the mean (45%). No differences in precision and accuracy were found between methods, except for TPM, for which gas chromatography showed poorer accuracy compared with immunoassay and gas chromatography-mass spectrometry. CONCLUSIONS: With the notable exception of TGB, interlaboratory variability in the determination of new AEDs was comparable to that reported with older-generation agents. Poor assay performance is related more to individual operators than to the intrinsic characteristics of the method applied. Participation in an EQAS scheme is recommended to ensure adequate control of assay variability in therapeutic drug monitoring. PMID- 12581229 TI - Talampanel, a new antiepileptic drug: single- and multiple-dose pharmacokinetics and initial 1-week experience in patients with chronic intractable epilepsy. AB - PURPOSE: Talampanel (LY300164), a potent and selective alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionate (AMPA)-receptor antagonist, is a potential new antiepileptic drug (AED). This study examines the single- and multiple-dose pharmacokinetics, safety, and tolerability of talampanel in patients with intractable epilepsy and assesses the potential for pharmacokinetic interaction. METHODS: Eleven of 14 patients entered into the study completed. Fourteen patients were evaluated for safety, 13 patients were used in the single-dose, and 11 patients in the multiple-dose pharmacokinetic analysis. Each patient initially received a single 35-mg dose of talampanel followed by the measurement of pharmacokinetic profiles. A 21-day t.i.d. dosing regimen was then determined for each patient based on his or her initial pharmacokinetic profile. Adverse events were recorded by patients or their carers. RESULTS: After oral ingestion, talampanel was rapidly absorbed, with maximal plasma concentrations achieved within 1-3 h. Talampanel concentrations in patients taking enzyme-inducing AEDs were 50% lower than those seen in healthy volunteers. Mean talampanel t1/2 values were 3.0 h compared with 4.2 h in healthy volunteers. After multiple-dose and steady-state, talampanel t1/2 values were increased to 5.6 h Talampanel and valproic acid (VPA) appear to inhibit each other's metabolism mutually. Talampanel had no effect on plasma concentrations of other AEDs. Multiple-dose talampanel administration was associated with nonlinear pharmacokinetics. No serious adverse events were reported; the most frequently reported being dizziness, ataxia, drowsiness, and headaches CONCLUSIONS: Talampanel dosing strategies may be reliant on concomitant AED medication, as enzyme-inducing AEDs enhance, whereas VPA inhibits its metabolism. Talampanel was well tolerated, although adverse events occurred at lower doses compared with those in healthy subjects, probably because of the additive effect of concomitant AEDs. PMID- 12581230 TI - Brain diffusion after single seizures. AB - PURPOSE: Diffusion-weighted magnetic resonance imaging (DWI) after focal status epilepticus has demonstrated focal alterations of the apparent diffusion coefficient (ADC) in the epileptogenic zone. We hypothesized that localized dynamic alterations of brain diffusion during the immediate postictal state will be detectable by serial DWI and correlate with the epileptogenic zone. METHODS: Nine adult patients (four men, five women) with medically intractable epilepsy were prospectively examined with a total of 25 DWI scans taken 2-210 min after a seizure. RESULTS: The interictal ADC was significantly (p < 0.05) elevated in the ictogenic hippocampus in all patients with temporal lobe epilepsy. The following postictal changes of the ADC were seen: (a) decreases by maximally 25-31%, which were most pronounced in the epileptogenic zone (n = 2); (b) generalized ADC changes after generalized seizures (n = 1) or prolonged complex partial seizures (n = 2); (c) no major changes after short-lived seizures or if the time to first DWI scan was >15 min or both (n = 3); and (d) widespread bilateral ADC increases after a flumazenil-induced seizure (n = 1). CONCLUSIONS: ADC changes seen during serial postictal DWI are complex and appear to reflect origin and spread of the preceding seizure. A delineation of the epileptogenic zone appears to be possible only in complex-partial seizures of >60 s duration that do not secondarily generalize. PMID- 12581231 TI - Psychogenic, nonepileptic seizures associated with video-EEG-verified sleep. AB - PURPOSE: Nonepileptic seizures (NES) are expressions of a psychiatric disease state, usually conversion disorder, that mimic epileptic seizures (ES) but are not associated with the neurophysiologic changes of epilepsy. Conversion has not been demonstrated to emerge from the sleeping state. Emergence out of sleep is usually considered a virtual exclusion criterion for NES, signifying the presence instead of ES. We sought to test this hypothesis. METHODS: We retrospectively reviewed the video-EEG of all patients admitted to our epilepsy unit over a 3 year period, who were suspected of manifesting NES. We examined the relation between NES and the patients' sleep/wake state in this cohort. Two epileptologists blindly reviewed an intermixture of cases suspected to represent NES emerging out of sleep, together with control cases. Classification of each case was made independently. RESULTS: We found that in a small minority of patients (<1%), NES began either while the patient was sleeping, or within several seconds of arousal, well before a plan to simulate a seizure could likely have been formulated in the wakeful state. CONCLUSIONS: In some cases, NES are not the product of the awake mind, but rather represent a psychiatric condition that can be manifest in sleep. PMID- 12581232 TI - Epileptogenicity of focal malformations due to abnormal cortical development: direct electrocorticographic-histopathologic correlations. AB - PURPOSE: Malformations due to abnormal cortical development (MCDs) are common pathologic substrates of medically intractable epilepsy. The in situ epileptogenicity of these lesions as well as its relation to histopathologic changes remains unknown. The purpose of this study was to correlate the cellular patterns of MCDs with the expression of focal cortical epileptogenicity as assessed by direct extraoperative electrocorticographic (ECoG) recordings by using subdural grids. METHODS: Fifteen patients with drug-resistant focal epilepsy due to pathologically confirmed MCD who underwent subdural electrode placement for extraoperative seizure localization and cortical mapping between 1997 and 2000 were included in the study. Areas of interictal spiking and ictal onset patterns were identified and separated during surgery for further pathologic characterization (cellular and architectural). Three pathologic groups were identified: type I; architectural disorganization with/without giant neurons, type IIA; architectural disorganization with dysmorphic neurons, and type IIB; architectural disorganization, dysmorphic neurons, and balloon cells (BCs). The focal histopathologic subtypes of MCDs in cortical tissue resected were then retrospectively correlated with in situ extraoperative ECoG patterns. RESULTS: Cortical areas with histopathologic subtype IIA showed significantly higher numbers of slow repetitive spike pattern in comparison with histopathologic type I (p = 0.007) and normal pathology (p = 0.002). The ictal onset came mainly from cortical areas with histopathologic type IIA (nine of 15 patients). None of the seizures originated from neocortical areas that showed BC containing MCD (type IIB). CONCLUSIONS: This study shows that areas containing BCs are less epileptogenic than are closely located dysplastic regions. These results suggest a possible protective effect of BCs or a severe disruption in the neuronal networks in BCs containing dysplastic lesions. Further studies are needed to elucidate the nature and the potential role(s) of balloon cells in MCD induced epileptogenicity. PMID- 12581233 TI - Prognostic implications of seizure recurrence in the first year after anterior temporal lobectomy. AB - PURPOSE: Seizure recurrence after anterior temporal lobectomy (ATL) incites concerns of whether seizures will eventually be successfully controlled. Our study evaluated the prognostic significance of seizure recurrence in the first year after ATL. METHODS: The postoperative courses of 175 consecutive patients who had undergone ATL and had > or =2 years of follow-up were studied. Recurrence was considered early if the first seizure occurred within 7 days after ATL and late if it occurred >7 days after ATL. Recurrent seizures were considered provoked when precipitating factors were present, such as interruption of antiepileptic drug (AED) intake. Subsequent outcome was determined at terminal follow-up. RESULTS: Percentage of excellent outcome was comparable between patients whose initial recurrent seizures were auras or simple partial seizures and patients without seizure recurrence in the first year (86.7 vs. 93.1%; p > or = 0.05). However, percentage of excellent outcome was less when the initial recurrent seizure was complex partial, either with or without secondary generalization (44.8%; p < or = 0.01). Outcome was not different between early and late seizure recurrence (excellent in 41.7 vs. 55.7%; p > or = 0.05). Nonetheless, patients with either early or late seizure recurrence were less likely to have excellent outcome than were patients with no seizure recurrence in the first year (p < or = 0.001). Percentage of excellent outcome was best when patients were seizure free in the first year (93.1%), intermediate when initial recurrent seizure was provoked (72.0%), and worst when unprovoked (27.8%) (p < or = 0.001). CONCLUSIONS: In the first postoperative year, the type of initial recurrent seizure, whether aura or complex partial and whether provoked or unprovoked, is associated with long-term prognosis in seizure control after ATL. The timing of the initial seizure recurrence is not as important. PMID- 12581234 TI - A study of 43 patients with panayiotopoulos syndrome, a common and benign childhood seizure susceptibility. AB - PURPOSE: To determine prevalence, clinical, EEG features, and prognosis of Panayiotopoulos syndrome and to examine the proposition that clinical manifestations are more important than EEG findings. METHODS: We analyzed retrospectively the clinical and EEG records of 1,340 children with one or more focal seizures seen in the last 18 years, supplemented with a prospective study from 1998. Panayiotopoulos syndrome was defined by clinical criteria, mainly ictal emesis, irrespective of EEG findings. RESULTS: We analyzed 43 of 90 patients with Panayiotopoulos syndrome who were seizure free >2 years. Girls predominated. Mean age at first seizure was 5 years. Seizures consisted mainly of autonomic manifestations; ictal emesis was often the first symptom, culminating in vomiting in 86%. Of nonautonomic manifestations, lateral eye deviation was the most common; visual symptoms were exceptional. Impairment of consciousness ensued in all seizures, half of which ended with hemi or generalized convulsions. Nearly 46.5% of cases had at least one seizure >30 min, constituting autonomic status epilepticus. Seizures during sleep (84%) were more common than those in wakefulness. EEG showed occipital spikes in 29 patients. Of the other 14 cases, five had extraoccipital abnormalities or brief generalized discharges, and nine had normal awake and sleep EEG. Prognosis was excellent. All 43 children have been free of seizures for > or =2 years, 53% having a single seizure, and 47%, an average two to three seizures. CONCLUSIONS: Panayiotopoulos syndrome is common and needs wider recognition. EEG shows occipital or extraoccipital abnormalities, is normal in one third of patients, and does not determine clinical manifestations or prognosis, which is excellent despite the high prevalence of lengthy seizures. PMID- 12581235 TI - A detailed analysis of symptomatic posterior cortex seizure semiology in children younger than seven years. AB - PURPOSE: To analyze the semiology of seizure onset and evolution in young children with posterior cortex epilepsy (PCE), compare this with adult reports, and assess age-related differences. METHODS: We videotaped and analyzed 110 seizures from 18 patients with PCE, aged 3-81 months. All had a good prognosis after posterior epileptogenic zone removal. Ictal events were categorized by behavioral, consciousness, autonomic, and sensory features, as well as motor patterns, which included myoclonic, tonic, clonic, unclassified motor seizures, and epileptic spasm. A time-scaled data sheet was developed to record each epileptic event as onset, very early, early, or late manifestation. RESULTS: Patients had a high seizure frequency with < or =100 attacks/day; one third of them showed a cluster tendency. The mean duration of seizures was 67 s. The most common seizure components were motor manifestations (with myoclonic and tonic seizures), but psychomotor (automotor), hypomotor attacks, and isolated auras also were frequently observed. Clinical seizure spread was frequent; auras and visual sensory signs were difficult to record in this age. Typical phenomena during seizures included behavioral changes, ictal vocalization, smile, flush, head nod, oculomotor features, and late-appearing oral automatisms, whereas hypermotor and secondarily generalized tonic-clonic seizures were not seen. CONCLUSIONS: Our results suggest that PCE in infants and young children is very heterogeneous but shows important age-related features. Compared with adults, children with PCE have shorter but more frequent seizures; they rarely report aura or visual sensory signs, only sporadically develop hypermotor and secondarily generalized tonic-clonic seizures, whereas ictal smile, flush, head nod, and behavioral change are typical features at this age. Because of frequent subtle ictal phenomena, long-term video-EEG monitoring is a useful diagnostic tool with infants and young children with PCE. PMID- 12581236 TI - Behavioral problems in children with newly diagnosed idiopathic or cryptogenic epilepsy attending normal schools are in majority not persistent. AB - PURPOSE: To investigate relevant adults' perceptions of behavioral problems in 66 children with newly diagnosed "epilepsy only" and in 63 healthy gender-matched classmates. METHODS: Parents' and teachers' perceptions of the children's behavioral problems were quantified by using the Total Problem score of the Child Behavior Checklist (CBCL) and the Teacher's Report Form (TRF), after correction for epilepsy-related item ambiguity. Questionnaires were filled out immediately after diagnosis and 3 and 12 months later. Relations with demographic and educational variables, school attitudes, and interview-derived prior adversities were analyzed. RESULTS: As a group, children with cryptogenic rather than those with idiopathic epilepsy have more behavioral problems than do healthy classmates. Family troubles and long-standing behavioral and learning problems are associated with more behavioral problems. The child's adaptation to the adversity of epilepsy onset is important. No adverse effect of antiepileptic drug (AED) treatment was found. Although the percentages of patients with clinically relevant (mean of controls + 2 SD) behavioral problems are consistently 25% (parents) and 22% (teachers), at each assessment, different children contribute to these percentages. In not a single child did parents and teachers agree on the presence of clinically relevant behavioral problems. CONCLUSIONS: (a) Behavioral problems are common in "epilepsy only," but are not persistent. (b) Agreement between parent's and teacher's perceptions of behavior is low. (c) Behavioral problems are perceived to occur already in the earliest stage of the disease. PMID- 12581237 TI - A multicenter study on the prevalence of psychiatric disorders among new referrals for epilepsy in Japan. AB - PURPOSE: To investigate the prevalence rate and risk factors of psychiatric disorders among new referrals for epilepsy, a multicenter study was conducted by using the International League Against Epilepsy (ILAE) criteria for epilepsy and the ICD-10 criteria for psychiatric disorders. METHODS: From April 2000 to March 2001, 398 patients with epilepsy, who were referred to nine neuropsychiatric outpatient clinics specialized for epilepsy in the Tokyo metropolitan area, were evaluated by using a newly developed five-axis classification scheme. RESULTS: Forty-two percent of the subjects showed a psychiatric disorder. Twenty-four percent of the total showed psychiatric disorders, including neurotic disorders in 8%, psychotic disorders in 7%, and affective disorders in 1%. In addition, 23% of the total showed mental retardation, and 18% showed personality disorders. A logistic regression analysis revealed that the three risk factors for a psychiatric disorder were mental retardation, temporal lobe epilepsy (as opposed to other subtypes), and a high seizure frequency. CONCLUSIONS: The presence of mental retardation was the primary risk factor for developing a psychiatric disorder, especially a schizophrenia-spectrum disorder. The type of epilepsy alone is not a strong predictor of psychiatric illness, and intractable temporal lobe epilepsy with a high seizure frequency is accountable for the link between the epilepsy and the psychiatric illness. PMID- 12581238 TI - Cross-cultural differences in levels of knowledge about epilepsy. AB - PURPOSE: To study how much people with epilepsy in Europe know and understand about their condition and how this might affect their lives. METHODS: Clinical, demographic, psychosocial details and information assessing knowledge were collected by using self-completion questionnaires mailed to members of epilepsy support groups. RESULTS: Data were collected from 6,156 people with epilepsy from ten European countries. There were significant between-country differences in all variables considered. Overall levels of knowledge were acceptable when measured by the epilepsy knowledge questionnaire (EKQ, medical items). However, there were some gaps in knowledge, particularly in issues relating to medication and cause of epilepsy. CONCLUSIONS: This is the largest study of its kind to date. Results clearly highlighted that levels of knowledge differed significantly between countries. Overall, people with epilepsy are reasonably well informed about epilepsy, although some gaps in knowledge were evident. PMID- 12581239 TI - Ictal urinary urge: further evidence for lateralization to the nondominant hemisphere. AB - PURPOSE: To determine the lateralizing value of ictal urinary urgency. METHODS: A retrospective database search was performed for patients with ictal urinary urgency admitted to the Epilepsy Monitoring Unit at the Cleveland Clinic between 1994 and 2001. RESULTS: Six patients were identified; intracarotid amytal test demonstrated left hemispheric speech dominance in five cases. The sixth patient continued to speak during right temporal seizures. EEG and imaging data supported right temporal or frontotemporal epilepsy in all six cases. Two patients were seizure free after focal right hemispheric resection. CONCLUSIONS: Ictal urinary urge appears to be a lateralizing sign for nondominant temporal lobe epilepsy. PMID- 12581240 TI - Bilateral Rasmussen encephalitis: postmortem documentation in a five-year-old. AB - A case of Rasmussen encephalitis with bilateral involvement and onset at age 2 years is reviewed. Rasmussen encephalitis is a rare progressive disease that causes intractable seizures, cognitive decline, and inflammatory changes in the brain. The neurologic involvement is characteristically unilateral. Bilateral involvement in this case was suspected within 5 months of presentation and was confirmed by bilateral frontal lobe biopsies. The severity and progression of the disease in this case was remarkable and resulted in the patient's death by age 5 years, despite numerous attempted therapeutic interventions. Autopsy findings confirmed the diagnosis of Rasmussen encephalitis with bilateral involvement. PMID- 12581241 TI - Mesial temporal sclerosis in acute childhood leukemias. AB - PURPOSE: Although seizures are relatively common in acute childhood leukemias, evolution into epilepsy is rare. METHODS: We describe three patients with acute leukemias who received chemotherapy. One patient also received cranial irradiation. RESULTS: All three developed recurrent complex partial seizures after initiation of chemotherapy. Initial neuroimaging performed in two patients was normal. Subsequent neuroimaging in all three revealed mesial temporal sclerosis. CONCLUSIONS: The association of mesial temporal sclerosis in acute childhood leukemias has not been previously described and may be secondary to antileukemic treatment and recurrent seizures. PMID- 12581242 TI - Attention deficits are not characteristic of school children with newly diagnosed idiopathic or cryptogenic epilepsy. PMID- 12581246 TI - Low prevalence of incidentally discovered and early-stage esophageal cancers in a 30-year autopsy study. AB - Autopsy studies provide information that may guide future patient management. This study analyzed autopsy findings in patients with esophageal cancers, with emphasis on the prevalence of incidentally diagnosed esophageal cancer, histologic subtypes, early-stage lesions, and any associated pathology. Autopsies detected 346 patients (306 men; 40 women) with esophageal carcinomas during a 30 year period, constituting an overall prevalence of 3.4%. Out of these patients, 30 (8.7%) were incidentally discovered at autopsy. Squamous cell cancers were found in 336 (97.1%) patients, small cell cancers in nine (2.7%), and adenocarcinomas in one (0.3%). Stage distributions were stage I in seven patients (2%), stage II in 49 (14%), stage III in 121 (35%), and stage IV in 169 (49%). Isolated dysplasia or carcinoma-in-situ were not found. Comparing with symptomatic patients, patients with incidental cancers were older, had higher frequency of small cell carcinomas, and lower T-stage. The prevalence of incidentally diagnosed esophageal carcinomas and early-stage carcinomas were low. Unusual histologic subtypes may be found. PMID- 12581247 TI - Cervical lymphadenectomy is beneficial for patients with carcinoma of the upper and mid-thoracic esophagus. AB - The role of cervical lymphadenectomy for thoracic esophageal cancer is controversial. This study evaluated the impact of cervical lymphadenectomy on the cervical lymph node metastasis (LNM) and survival rates of patients with esophageal cancer. We analyzed 199 patients who received radical esophagectomy with three-field lymphadenectomy. The overall 5-year survival rate was 49.4%. Cervical LNM was found in 36 (18.1%) out of the 199 patients. The 5-year survival rates of the patients with cervical LNM from upper and mid-esophageal cancers were 71.4% and 35.9%, respectively. However, none of the patients with cervical LNM from lower esophageal cancer survived more than 4 years after esophagectomy. The overall survival of patients with five or more metastatic nodes (5.9%) was significantly worse than that of patients with less than five positive nodes (45.5%). Cervical lymphadenectomy is beneficial for patients with carcinoma of the upper and mid-thoracic esophagus, and with less than five positive nodes. PMID- 12581248 TI - Preoperative chemoradiotherapy in cancer of the thoracic esophagus. AB - Surgery with or without adjuvant radiotherapy (RT) is the standard treatment of esophageal cancer. Preoperative radio- and chemotherapy (CT) have been introduced to improve prognosis. We report a phase II prospective non-randomized trial of preoperative RT (42 Gy/25) plus CT (cisplatin 20 mg/mq/day plus 5-fluorouracil 600 mg/mq/day, 1-5 weeks) for the treatment of thoracic esophageal cancer. From 1993, 50 patients were enrolled (40 men and 10 women, mean age 57 years, range 30 75 years). Squamous cell carcinoma accounted for 90% of cases; 10% were adenocarcinoma. Downstaging of the disease was obtained in 77.3% of cases; there were 13 (29.5%) complete responses (CR) and 21 (47.7%) partial responses (PR). Median survival was 28 and 25 months, respectively, for CR and partial response (PR) plus stable disease (SD) and progressive disease (PD) (P = 0.05). Progressive-free median survival was 22 and 17 months, respectively, for CR and PR + SD + PD (P = 0.08). Multimodal treatment of esophageal cancer showed promising results, although not significant, in terms of survival and disease progression for patients achieving a complete pathologic response. PMID- 12581250 TI - Prevalence of intestinal metaplasia according to the length of the specialized columnar epithelium lining the distal esophagus in patients with gastroesophageal reflux. AB - The diagnosis of Barrett's esophagus is based on the presence of intestinal metaplasia (IM) at the distal esophagus. The aim of this study was to determine the prevalence of IM in patients with symptoms of gastroesophageal reflux in whom endoscopically a segment of distal esophagus was covered by columnar epithelium (CE). In a prospective, descriptive and transversal study, 492 patients (33%) from 1480 patients with gastroesophageal reflux, in whom endoscopic evaluation demonstrated the presence of a short-segment CE measuring less than 3 cm or a long-segment CE measuring more than 3 cm, were evaluated. From each patient, several biopsy specimens were taken, which were stained with hematoxylin-eosin and Alcian blue pH 2.5. Out of 492 cases, 421 patients (86%) presented with a short-segment CE and 71 patients (14%) had a long-segment CE. Among these 71 cases, 38 had a 3-6 cm-length CE, 21 patients had a 6.1-10 cm-length CE and 12 patients had CE more than 10.1 cm in length. Endoscopic short-segment CE was six times more frequent than long-segment CE. The prevalence of IM was 35% among patients with short-segment CE and increased progressively according to the length of CE, being 100% in patients with > 10 cm in length. Therefore, true short-segment BE was three times more frequent during endoscopic studies than long-segment BE. Dysplasia in the metaplastic epithelium also increased parallel to the length of the CE. True BE (presence of IM at the columnar epithelium lining the distal esophagus), was present in 13.6% of all patients with symptoms of gastroesophageal reflux submitted to endoscopic evaluation. Short-segment BE is three times more frequent than long-segment BE, and endoscopic and bioptic evaluation is fundamental in all cases with gastroesophageal reflux who exhibit some segment of the distal esophagus lined by columnar epithelium, even if it is > or = 1 cm long. PMID- 12581249 TI - Loss of Bcl-2 expression in Barrett's dysplasia and adenocarcinoma is associated with tumor progression and worse survival but not with response to neoadjuvant chemoradiation. AB - Esophageal adenocarcinoma arising on a background of Barrett's esophagus is increasing in incidence. A molecular understanding of both the progression of Barrett's esophagus and the factors determining the response of adenocarcinoma to neoadjuvant therapy is required, and this study focused on the role of proteins regulated by the bcl-2 family of genes, which are important regulators of programmed cell death (apoptosis). In total, 48 patients (36 men, 12 women) with Barrett's adenocarcinoma were studied. All patients received preoperative chemoradiotherapy followed by surgery. Bcl-2, bax and bcl-x protein expression were detected by standard avidin-biotin peroxidase method. Bcl-2, bax and bcl-x expression were detected in 84%, 80%, and 76%, respectively, of normal squamous mucosa. An increasing degree of dysplasia in Barrett's mucosa both before and after chemoradiotherapy was significantly associated with a reduction of bcl-2 expression (P = 0.03 and 0.009, respectively). Bcl-2 expression was significantly associated with tumor differentiation (P = 0.03) and a trend towards earlier T stage (P = 0.08), but not with nodal status. Pre-therapeutic bcl-2, bax and bcl-x protein expression (27%, 75%, and 87.5%, respectively) were not associated with tumor response or resistance to therapy. Bcl-2-positive patients had a significantly improved survival compared with bcl-2-negative tumors. A significant reduction of bcl-2 expression is associated with the progression of Barrett's mucosa to adenocarcinoma. Bcl-2 expression was associated with improved survival. Preoperative chemoradiotherapy induces expression of bax and bcl-x protein. The pretreatment expression of bcl-2 and related proteins did not predict response or resistance to neoadjuvant chemoradiotherapy, suggesting that regulators of apoptosis alone do not determine the response of Barrett's adenocarcinoma to neoadjuvant therapy. PMID- 12581251 TI - Intestinal metaplasia in the distal esophagus and correlation with symptoms of gastroesophageal reflux disease. AB - Barrett's esophagus is a metaplastic condition that occurs in patients with gastroesophageal reflux disease (GERD) and its importance lies in its potential to develop adenocarcinoma of the esophagus. The diagnosis of Barrett's esophagus is based on finding of intestinal metaplasia of at least 3 cm of the distal esophagus. The diagnosis of intestinal metaplasia of less than 3 cm of the distal esophagus is controversial, regarding implications with GERD, adenocarcinoma, and Helicobacter pylori. The aims of the study were to determine the prevalence of intestinal metaplasia in the distal esophagus in patients with short segments of esophageal columnar-appearing mucosa (less than 3 cm), diagnosed endoscopically, in two groups of patients, with and without symptoms of GERD. In total, 97 patients were examined, with endoscopic finding of esophageal columnar-appearing mucosa less than 3 cm. From the total, 52 patients had symptoms of GERD and 45 patients were without these symptoms. These patients were subjected to distal esophageal biopsies obtained immediately below the epithelial transition. The biopsies were stained with hematoxylin-eosin and alcian blue at pH 2.5. Urease test for H. pylori detection in two fragments of gastric antrum was carried out. The presence of intestinal metaplasia in the distal esophagus was diagnosed in 16 (30.8%) patients in the GERD group and 12 (26.7%) patients without GERD symptoms. No statistical differences were observed (P = 0.82; 95% CI: 0.61-2.17). The variables sex, mean age and positivity for H. pylori did not show statistical differences. This study diagnosed high prevalence of intestinal metaplasia in the distal esophagus with columnar-appearing mucosa, less than 3 cm, with no statistical differences in the two groups studied with and without GERD symptoms. PMID- 12581252 TI - Intrasphincteric botulinum toxin injection in the treatment of chagasic achalasia. AB - According to the WHO, 16-18 million people in Central and South America are infected by Trypanosoma cruzi. Chagasic achalasia affects between 7.1% and 10.6% of the population. The aim of this study was to evaluate the effects of Botox injections in the clinical response and esophageal function of patients with dysphagia due to chagasic achalasia. In total, 24 symptomatic patients with chagasic achalasia were randomly chosen to receive Botulinum Toxin (BT) or saline injected by endoscopy in the lower esophageal sphincter (LES). Patients were monitored with a clinical score of dysphagia and an objective assessment (esophagograms, scintillography, manometry, and nutritional assessment) for a period of 6 months. Clinical improvement of dysphagia was statistically significant (P < 0.001) in patients receiving BT when compared with the placebo. There was no significant difference in the placebo group regarding clinical score, LES basal pressure and esophageal emptying time. Esophageal emptying time in the toxin group was significantly lower than in the placebo (P=0.04) after 90 days. There were non-significant increases in esophageal emptying of 25.36% and 17.39%, respectively, at 90 and 180 days, in the BT group (P=0.266). Gender, age, and baseline LES pressure did not influence the response to BT. Our data strongly suggests that intrasphincteric injection of BT in LES is clinically effective in the treatment of chagasic achalasia. PMID- 12581254 TI - Isolated choroidal metastasis from primary adenocarcinoma of the distal esophagus: a case report. AB - We present a case of a 41-year-old male patient who, 1 year after transhiatal esophagectomy and transmediastinal gastroplasty for an adenocarcinoma of the distal esophagus, presented with an isolated metastasis in the choroids of the left eye. Because of pains caused by secondary glaucoma, enucleation of the left eyeball had to be performed as the treatment of choice. At 1 year after surgery of the eye, the secondaries in both adrenal glands were revealed. Despite the applied chemotherapy, the patient died with signs of generalized disease 3 years after the initial surgery. PMID- 12581253 TI - Isolated esophageal atresia and perinatal risk factors. AB - Esophageal atresia (EA) is a relatively rare alimentary tract congenital anomaly. We studied perinatal risk factors of 90 live birth cases of isolated EA with (40 cases) or without (50 cases) tracheal fistula. We found sex ratios of 1.43 for total EA cases and 1.5 for both subgroups of EA with or without tracheal fistula. Evaluation of parity showed statistically significant values for low parity and for high multiparity for EA with and without tracheal fistula. Birth weight evaluation revealed statistically significant values for both subgroups. We did not find statistical significance both for maternal age and gestational age. The etiology of esophageal atresia is complex and heterogeneous and within each subgroup, apparently different etiologies may exist, resulting in differences in epidemiologic characteristics. Moreover, some causes linked to genetics and gene environment interaction may be involved. We think that parity and low birth weight can be considered as risk factors for EA, and an accurate evaluation of reproductive history can be useful for the provision of genetic and perinatal counseling. PMID- 12581255 TI - Spontaneous expulsion of a mediastinal lymph node in carcinoma of the esophagus. AB - A case of spontaneous expulsion of a mediastinal lymph node, which developed during the follow up of a patient with carcinoma of the esophagus is presented. To the best of our knowledge, no such instance of natural extrusion of mediastinal lymph node has been reported in the literature. PMID- 12581258 TI - Medical communication and diabetes. PMID- 12581256 TI - Lichen planus esophagitis: report of three patients treated with oral tacrolimus or intraesophageal corticosteroid injections or both. AB - Clinically significant involvement of the esophagus is uncommon in patients who have lichen planus, a common disorder of squamous epithelium. In three patients who had oral, cutaneous, and esophageal lichen planus, endoscopic intralesional esophageal injection of corticosteroids (in all three patients) and oral tacrolimus (FH506) (in two patients) resulted in improvement in dysphagia, a less frequent need for dilation, and improvement in esophageal inflammation. PMID- 12581259 TI - The pathogenesis and management of painful diabetic neuropathy: a review. AB - Painful diabetic neuropathy has always been a challenging complication of diabetes mellitus. Emerging theories suggest that early dysaesthesia associated with painful neuropathy may act as a marker for the development of the 'at risk' foot, allowing preventative clinical strategies to be undertaken. The mechanisms of neuropathic pain are complex. The authors' intentions are to help members of the diabetes care team better understand and appreciate the diverse symptoms reported by patients. The various treatments available for painful neuropathy are discussed in detail. Robust comparative studies on such treatments are, however, unavailable and the authors have designed a logical approach to management based on best current evidence and their own clinical experience. PMID- 12581260 TI - Factor analysis of the metabolic syndrome among elderly Koreans--the South-west Seoul Study. AB - AIMS: To examine the relationship between the components of the metabolic syndrome and to explore whether insulin resistance unifies the clustering of components of the metabolic syndrome among urban elderly Koreans using exploratory factor analysis. METHODS: We included 1314 non-diabetic subjects over the age of 60 years, selected from a cross-sectional study, which was conducted in 1999 in Seoul, Korea. Factor analysis was carried out using the principle components analysis with Varimax orthogonal rotation of the components of the metabolic syndrome. RESULTS: We found four major factors of cardiovascular disease risk variables in our study subjects. Impaired glucose tolerance, dyslipidaemia, hypertension and obesity aggregated as the major domain. Obesity and dyslipidaemia variables were closely related and loaded on the same factor. However, hypertension was not linked closely with other factors of the metabolic syndrome. CONCLUSIONS: Insulin resistance is not the only contributor to the metabolic syndrome among urban elderly Koreans. Although the components of the metabolic syndrome were closely related, the finding of more than one factor suggests that more than one pathophysiological mechanism underlies full expression of the metabolic syndrome among elderly Koreans. PMID- 12581261 TI - Foot complications in Type 2 diabetes: an Australian population-based study. AB - AIMS: To determine the prevalence and risk factors for neuropathy and peripheral vascular disease (PVD) in the Australian diabetic population and identify those at high risk of foot ulceration. METHODS: The Australian Diabetes Obesity and Lifestyle study included 11 247 adults aged >or= 25 years in 42 randomly selected areas of Australia. Neuropathy and PVD were assessed in participants identified as having diabetes (based on self report and oral glucose tolerance test), impaired fasting glucose, impaired glucose tolerance and in a random sample with normal glucose tolerance (total n = 2436). RESULTS: The prevalence of peripheral neuropathy was 13.1% in those with known diabetes (KDM) and 7.1% in those with newly diagnosed (NDM). The prevalence of PVD was 13.9% in KDM and 6.9% in NDM. Of those with diabetes, 19.6% were at risk of foot ulceration. Independent risk factors for peripheral neuropathy were diabetes duration (odds ratio (95% CI) 1.73 (1.33-2.28) per 10 years), height (1.42 (1.08-1.88) per 10 cm), age (2.57 (1.94-3.40) per 10 years) and uric acid (1.59 (1.21-2.09) per 0.1 mmol/l). Risk factors for PVD were diabetes duration (1.64 (1.25-2.16) per 10 years), age (2.45 (1.86-3.22) per 10 years), smoking (2.07 (1.00-4.28)), uric acid (1.03 (1.00 1.06) per 0.1 mmol/l) and urinary albumin/creatinine ratio (1.11 (1.01-1.21) per 1 mg/mmol). CONCLUSIONS: The prevalence of neuropathy and PVD was lower in this population than has been reported in other populations. This may reflect differences in sampling methods between community and hospital-based populations. Nevertheless, a substantial proportion of the diabetic population had risk factors for foot ulceration. PMID- 12581262 TI - A pilot study of the beneficial effects of amantadine in the treatment of painful diabetic peripheral neuropathy. AB - BACKGROUND: Current symptomatic treatments for painful peripheral neuropathy in diabetes have variable efficacy in individual patients. Amongst other chemical transmitters involved in pain reception, the N-methyl-D-aspartate (NMDA) subtype of excitatory amino acid receptor is involved in nociception. Amantadine was recently shown to act as a non-competitive antagonist of NMDA and may be effective in the treatment of neuropathic pain in patients with cancer. We have looked at the benefit of amantadine infusion in diabetic patients with painful peripheral neuropathy. METHODS: Seventeen patients with diabetes (nine men) completed this double-blind randomized crossover placebo-controlled trial of intravenous amantadine. The average age was 58.4 (sd 11) years, with duration of diabetes of 21.1 (8.7) years and duration of painful peripheral neuropathy symptoms of 29.1 (24) months. All analgesics except paracetamol were stopped for 4 weeks prior to the study. Infusions were carried out on a weekly basis with amantadine being administered intravenously as a single 200-mg infusion. The Neuropathy Symptom Score (NSS), together with visual analogue scales, were used to assess current pain intensity (VAS-P) pre-therapy and 1 week later VAS-P was repeated together with a visual analogue scale used to assess relief in pain (VAS R) and the Physicians Global Evaluation (PGE) score used to assess response to therapy. RESULTS: Pre-therapy, the NSS was 6.8 (6.3-7.4) at baseline, remaining unchanged at 6.6 (5.8-7.4) after placebo (P = 0.33), but fell to 4.6 (3.4-5.8) after amantadine (P = 0.003 vs. baseline and P = 0.02 vs. placebo). The baseline perception of pain was scored as 7.8 cm (7.3-8.3), with no difference following placebo, at 8.2 cm (7.7-8.6) (P = 0.34), but following amantadine it fell to 6.2 cm (4.9-7.8) (P = 0.01 compared with pre-therapy, P = 0.003 compared with placebo). The perception of relief from pain following placebo was only 0.2 (-0.2 to +0.6) but following amantadine was 10-fold better at 1.9 (0.8-3.1) (P = 0.016). The PGE assessment of pain relief was -0.3 (-0.5 to 0) for placebo and following amantadine was 0.8 (0.1-1.5) (P = 0.006). CONCLUSIONS: Our study has shown that intravenous amantadine is beneficial in reducing the pain of painful peripheral neuropathy, with an effect sustained for at least 1 week after an infusion. PMID- 12581263 TI - Impairment of cerebral autoregulation in diabetic patients with cardiovascular autonomic neuropathy and orthostatic hypotension. AB - AIMS: Impaired cerebrovascular reactivity and autoregulation has been previously reported in patients with diabetes mellitus. However, the contribution of cardiovascular diabetic autonomic neuropathy and orthostatic hypotension to the pathogenesis of such disturbances is not known. The purpose of this study was to evaluate cerebral blood flow velocity in response to standing in patients with diabetes and cardiovascular autonomic neuropathy with or without orthostatic hypotension. METHODS: We studied 27 patients with diabetes--eight had cardiovascular autonomic neuropathy and orthostatic hypotension (age 46.4 +/- 13.5 years, diabetes duration 25.0 +/- 11.0 years), seven had autonomic neuropathy without hypotension (age 47.3 +/- 12.7 years, diabetes duration 26.4 +/- 12.1 years), and 12 had no evidence of autonomic neuropathy (age 44.1 +/- 13.8 years, diabetes duration 17.1 +/- 10.2 years)-and 12 control subjects (age 42.6 +/- 9.7 years). Flow velocity was recorded in the right middle cerebral artery using transcranial Doppler sonography in the supine position and after active standing. RESULTS: Cerebral flow velocity in the supine position was not different between the groups studied. Active standing resulted in a significant drop of mean and diastolic flow velocities in autonomic neuropathy patients with orthostatic hypotension, while there were no such changes in the other groups. The relative changes in mean flow velocity 1 min after standing up were -22.7 +/- 16.25% in patients with neuropathy and orthostatic hypotension, +0.02 +/- 9.8% in those with neuropathy without hypotension, -2.8 +/- 14.05% in patients without neuropathy, and -9.2 +/- 15.1% in controls. CONCLUSIONS: Patients with diabetes and cardiovascular autonomic neuropathy with orthostatic hypotension show instability in cerebral blood flow upon active standing, which suggests impaired cerebral autoregulation. PMID- 12581264 TI - Improvements in insulin sensitivity and beta-cell function (HOMA) with weight loss in the severely obese. Homeostatic model assessment. AB - AIMS: To examine the effect of weight loss on insulin sensitivity and beta-cell function in severely obese subjects of varying glycaemic control. PATIENTS AND METHODS: Subjects were 254 (F:M 209:45) patients having adjustable gastric banding for severe obesity, with paired biochemical data from before operation and at 1-year follow up. The homeostatic model assessment method was used to calculate insulin sensitivity (HOMA%S) and beta-cell function (HOMA%B). Subjects were grouped by diabetic status and by pre-weight loss HbA1c. RESULTS: Initial mean (sd) weight and body mass index were 128 (26) kg and 46.2 (7.7) kg/m2, respectively, and at 1-year were 101 (22) kg and 36.4 (6.7) kg/m2. The percentage of excess weight lost (%EWL) was 44.3 (14)%. HOMA%S improved from 37.5 (16)% presurgery to 62 (25)% (P < 0.001). %EWL was the only predictor of HOMA%S improvement (r = 0.28, P < 0.001). Subjects with normal fasting glucose, impaired fasting glucose and Type 2 diabetes had a fall, no change and increase in HOMA%B, respectively. The improvement in HOMA%B in subjects with diabetes (n = 39) was inversely related to the time with diabetes (r = -0.36, P = 0.02). In non diabetic subjects the HOMA%S-HOMA%B relationship was favourably altered with weight loss, so that for any given HOMA%S there was an increase in HOMA%B (f = 11.8, P = 0.001). This improvement in HOMA%B was positively related to %EWL (r = 0.25, P = 0.019). DISCUSSION: There are beneficial changes in both insulin sensitivity and beta-cell function with weight loss. Modern laparoscopic obesity surgery may have an important early role in the management of Type 2 diabetes in obese subjects. PMID- 12581265 TI - Psychosocial factors and diabetes-related outcomes following diagnosis of Type 1 diabetes in adults: the Edinburgh Prospective Diabetes Study. AB - AIMS: To examine prospectively the relationships between psychosocial variables and diabetes-related outcomes in adults with newly diagnosed Type 1 diabetes. METHODS: A total of 84 adults (48 male) with a median (range) age of 30.8 (17-51) years with newly diagnosed Type 1 diabetes were recruited for the study. Shortly after initial diagnosis each participant's personality, cognitive ability, and recent psychiatric distress were assessed. At 4 months (n = 69) and at 12 months (n = 66) after diagnosis diabetes-related outcomes were measured, including each respondent's knowledge of diabetes, satisfaction with diabetes treatment and diabetes-related quality of life. Glycated haemoglobin (HbA1c) was recorded at each clinic attendance. RESULTS: Social class (Spearman's correlation r = -0.30 and -0.28, respectively, P < 0.05) and scores on the National Adult Reading Test (r = 0.38 and 0.36, respectively, P < 0.01) were consistently associated with knowledge of diabetes at 4 months and at 12 months after diagnosis. Hierarchical regression revealed that alcohol consumption recorded at diagnosis and knowledge of diabetes at 4 months were independent predictors of glycaemic control at 12 months (adjusted r2 = 0.16). Total scores on the Diabetes Treatment Satisfaction Questionnaire (DTSQ) at 12 months were significantly predicted by age at diagnosis (adjusted r2 = 0.08). High neuroticism at diagnosis was consistently associated with poorer self-reported diabetes quality of life at 4 months and at 12 months after diagnosis (rs between -0.30 and -0.39, P < 0.05). CONCLUSIONS: Long-standing psychosocial factors have a significant influence on self-reported outcomes during the 12 months following diagnosis of Type 1 diabetes but may not be reliable predictors of glycaemic control. Further follow-up is necessary to determine the longer-term predictors of objective (e.g. glycaemic control) and subjective (e.g. quality of life) indicators of coping in people with diabetes. PMID- 12581266 TI - Do plasminogen activator inhibitor (PAI-1) or tissue plasminogen activator PAI-1 complexes predict complications in Type 1 diabetes: the Pittsburgh Epidemiology of Diabetes Complications Study. AB - AIMS: To examine the predictive power of plasminogen activator inhibitor-1 (PAI 1) and the complexes it forms with tissue plasminogen activator (tPA-PAI-1) for the two major Type 1 diabetes (T1D) complications (coronary artery disease (CAD) and overt nephropathy) in the context of standard risk factors. METHODS: Observational prospective study of 454 participants with childhood onset (< 17 years) T1D, aged 18+ years at baseline. PAI-1 and tPA-PAI-1 were determined using ELISA methodology. Follow-up (6 years) was limited to 382 individuals for CAD and 294 individuals for overt nephropathy, after excluding baseline cases. Total, HDL and LDL-cholesterol, triglycerides, HbA1, blood pressure, body mass index (BMI), waist-hip ratio (WHR), leucocyte count, Beck depression score and fibrinogen were also examined. RESULTS: The 56 incident cases of CAD had marginally lower PAI-1 and higher tPA-PAI-1 levels compared with those free of CAD. However, marginally higher PAI-1 and significantly higher tPA-PAI-1 (P = 0.04) levels were seen in those who developed nephropathy. After controlling for age, both PAI-1 and tPA PAI-1 showed significant negative correlations with HDL-cholesterol, and positive correlations with triglycerides, WHR, HbA1 and fibrinogen. tPA-PAI-1 was also positively correlated with total and LDL-cholesterol. In multivariate analyses, neither PAI-1 nor tPA-PAI-1 was an independent predictor of CAD or overt nephropathy. CONCLUSIONS: These results suggest little association between PAI-1 and later CAD in patients with T1D. However, tPA-PAI-1 complexes may be involved in the pathogenesis of overt nephropathy. PMID- 12581267 TI - Comparison of patients' and healthcare professionals' beliefs about and attitudes towards Type 2 diabetes. AB - AIMS: To compare patients' and healthcare professionals' beliefs and attitudes towards diabetes. METHODS: One hundred and four healthcare professionals (nurses, dieticians, general practitioners) and 100 of their patients with Type 2 diabetes completed the Diabetes Attitude Scale, 3rd version (DAS-3). Healthcare professionals also answered questions about their provision of diabetes care. RESULTS: Healthcare professionals viewed Type 2 diabetes as more serious than their patients. Most of the healthcare professionals considered diabetes harder to treat than other chronic conditions and felt that they did not have adequate time and resources to treat their diabetes patients effectively. CONCLUSIONS: This study suggests that an important part of forming collaborative alliances with patients is to recognize the distinction between practitioner and patient perspectives in Type 2 diabetes. PMID- 12581268 TI - Screening for Type 2 diabetes: is it worthwhile? Views of general practitioners and practice nurses. AB - AIMS: Screening for Type 2 diabetes does not meet all the criteria for screening, yet is increasingly proposed. The views of health care professionals towards screening for Type 2 diabetes are not known. We set out to explore the attitudes and beliefs held by general practitioners (GPs) and practice nurses towards screening for Type 2 diabetes. METHODS: Semistructured interviews with 10 GPs and nine practice nurses in eight general practices in North-east England. Data collection and analysis proceeded in an iterative manner in accordance with grounded theory. RESULTS: Practitioners who perceive themselves to be most aware of the evidence base surrounding screening for Type 2 diabetes are least likely to support its introduction. Those who support screening for Type 2 diabetes hold a 'common sense' viewpoint that earlier detection must be better and believe that patients demand and respect early diagnosis. Practitioners acknowledge a lack of knowledge and skills to promote lifestyle changes. This deficiency is combined with workload issues, contributing towards a medicalized view of diabetes screening. CONCLUSIONS: The belief that screening for Type 2 diabetes is worthwhile is based not on evidence for the effectiveness of screening, but rather on a complex interaction of factors including perceptions of patient desires and an interplay of previous experience and evidence from other sources. Increased resources would be most likely to promote screening in primary care. PMID- 12581269 TI - Methicillin-resistant Staphylococcus aureus in the diabetic foot clinic: a worsening problem. AB - AIMS: To determine if there has been a change in the prevalence of pathogenic organisms in foot ulcers in diabetic patients in 2001 compared with our previous study in 1998. METHODS: A retrospective analysis of wound swabs taken from infected foot ulcers in diabetic patients attending the outpatient clinic in the Manchester Foot Hospital over a twelve-month period. A total of 63 patients with positive wound swabs were identified. RESULTS: Gram-positive aerobic bacteria still predominate (84.2%) and the commonest single isolate remains Staphylococcus aureus (79.0%) which is higher than we previously reported. MRSA was isolated in 30.2% of the patients which is almost double the proportion of MRSA-affected patients three years ago. This did not appear to be related to prior antibiotic usage. There was no increase in hospitalisation because of MRSA infection. CONCLUSIONS: The problem of MRSA continues to increase despite the precautions taken to prevent MRSA spread. There is a need for a multi-centre study looking into the prevalence of MRSA in diabetic foot ulcer and how this can be reduced in the diabetic foot clinic. PMID- 12581270 TI - The relationship between birth weight and maternal glycated haemoglobin (HbA1c) concentration in pregnancies complicated by Type 1 diabetes. AB - AIM: To examine the relationships between maternal HbA1c concentration at different time points and birth weight in pregnancies complicated by pre-existing Type 1 diabetes. METHODS: A comprehensive audit dataset was collected prospectively on all deliveries in Scotland to women with pre-existing Type 1 diabetes occurring between 1 April 1998 and 31 March 1999. Data items included HbA1c concentrations prior to conception and in each of the three trimesters of pregnancy, and birth weight. Relationships between standardized birth weight and HbA1c concentrations at each of the four time points were examined using correlation analysis. RESULTS: Standardized birth weight (Z scores) could be calculated for 203 of 208 singleton liveborn infants. HbA1c concentrations, standardized to correct for assay differences among hospitals, at different time points were available for between 134 (pre-pregnancy) and 192 (third trimester) cases. Standardized birth weight, relative to a reference population, showed a unimodal distribution, shifted to the right (mean, +1.57 sd). There was a significant negative correlation between pre-pregnancy HbA1c and birth weight (Spearman's R, -0.208; P = 0.016). There were no statistically significant correlations for other time points. CONCLUSIONS: Standardized birth weight scores of the infants of diabetic mothers are higher than those of a reference population. There is no simple relationship between maternal glycaemic status and birth weight, but there appears to be a paradoxical inverse relationship between pre-pregnancy glycaemic control and standardized birth weight. PMID- 12581271 TI - Tumour necrosis factor-alpha promoter gene polymorphism at - 308 (genotype AA) in Chinese subjects with Type 2 diabetes. PMID- 12581272 TI - Insulin-treated diabetes and driving in the UK. PMID- 12581273 TI - Continued rising trend of childhood Type 1 diabetes mellitus in Devon and Cornwall, England. PMID- 12581274 TI - High prevalence of diabetes and impaired fasting glycaemia in urban Nepal. PMID- 12581276 TI - The local lymph node assay: past, present and future. AB - The local lymph node assay (LLNA) was developed originally as a method for the identification of chemicals that have the potential to cause skin sensitization and allergic contact dermatitis. The assay is based on an understanding that the acquisition of contact sensitization is associated with, and dependent upon, the stimulation by chemical allergens of lymphocyte proliferative responses in skin draining lymph nodes. Those chemicals that provoke a defined level of lymph node cell (LNC) proliferation (a 3-fold or greater increase compared with concurrent vehicle controls) are classified as skin sensitizers. Following its original inception and development, the LLNA was the subject of both national and international interlaboratory collaborative trials, and of very detailed comparisons with other test methods and with human skin sensitization data. The assay has now been validated fully as a stand-alone test for the purposes of hazard identification. In recent years, there has been a growing interest also in the use of the LLNA to assess the potency of contact allergens and in risk assessment. There is reason to believe that the extent of skin sensitization achieved is associated with the vigour of LNC proliferation induced in draining nodes. Given this relationship, the relative potency of skin sensitizing chemicals is measured in the LLNA by derivation of an EC3 value, this being the concentration of chemical required to provoke a 3-fold increase in the proliferation of LNC compared with controls. Experience to date indicates that relative potency as determined using this approach correlates closely with what is known of the activity of skin sensitizing chemicals in humans. In this article, we review the development, evaluation and validation of the LLNA for the purposes of hazard identification, and the more recent application of the method for evaluation of potency in the context of risk assessment. In addition, we consider what new applications and modifications are currently being investigated. PMID- 12581277 TI - Occupational irritant contact dermatitis from synthetic mineral fibres according to Finnish statistics. AB - Synthetic mineral fibres (i.e. man-made vitreous fibres, MMVF) are classified into glass filament, mineral wool (glass wool, rock wool and slag wool), refractory ceramic fibres, and fibres for special purposes. This paper analyses the data on occupational irritant contact dermatitis (ICD) caused by MMVF during 1990-99 in Finland according to the Finnish Register of Occupational Diseases (FROD). A total of 63 cases from MMVF were reported. 56 were diagnosed as ICD, and 2 as allergic contact dermatitis, both from rock wool. 53 out of 63 cases were due to mineral wool or glass filaments; half of the cases, according to a rough estimate, were due to mineral wool and a half due to glass filaments used in lamination work. Carpenters, building workers and insulation workers have the highest risk of ICD from mineral wool. 4 cases in carpenters, 4 in building workers and 2 in insulation workers were reported from MMVF. For every 100 000 employed workers, only 1.6 cases of ICD in carpenters, 2.7 in building workers and 9.1 in insulation workers were annually due to MMVF, respectively. Mineral wool used in construction work, insulation, etc., cannot be considered to be a common cause of occupationally induced ICD. However, information on harmful skin effects of MMVF is useful to exposed persons in the prevention of the effects. PMID- 12581278 TI - Audit of Finn Chamber patch test preparation. AB - Reproducibility of patch test reactions is an important aspect of patch testing. Thin layer rapid use epicutaneous tests (TRUE Test) are preloaded with allergen, whereas Finn Chambers are empty discs that have allergen applied from a syringe. The amount of allergen applied to each Finn Chamber is therefore potentially variable depending on technique. This may increase the risk of non reproducibility of reactions. We have performed an audit of patch test preparation in our department looking at weight variation of both individual Finn Chambers and 3 completed panels before and after allergen application. We found that the variance in weight was not significant for 2 of the 3 panels and single Finn Chambers with allergen in petrolatum. The variance in weight was significant for single Finn Chambers with allergen in solution and the third panel, which contains an allergen in solution. The clinical significance of these results is unclear. PMID- 12581279 TI - Cross-reactivity patterns to budesonide. AB - Allergic contact dermatitis from topical corticosteroids is not uncommon. Budesonide has been included in the European standard series as a marker for corticosteroid allergy, though little is known of its cross-reactivity with other corticosteroids. Twelve patients previously positive to budesonide on patch testing were given further patch and intradermal tests to a range of corticosteroids. Six patients previously negative to budesonide on patch testing were used as a control group. Budesonide cross-reacts with hydrocortisone-21 sodium phosphate and triamcinolone acetonide. Patients positive to budesonide should therefore avoid hydrocortisone and triamcinolone acetonide. Patch testing, unfortunately, is an inaccurate method of determining cross-reactivity patterns among corticosteroids. PMID- 12581280 TI - A clinical and patch test study of adult widespread eczema. AB - The purpose of this study was to investigate the role of contact hypersensitivity (CHS) in adult widespread eczema (WE) and to analyse the aetiology of WE. 108 consecutive adult WE patients were patch tested. 352 patients with suspected localized allergic contact dermatitis (ACD) tested in the same period served as a control. The average age of the patients with WE was significantly greater than the control group (47.6 +/- 15.0 v 39.2 +/- 14.9, p < 0.01, Student's t-test). More male patients were found in the WE group (42.6% vs 25.0%, p < 0.01, chi2 test). 71 (65.7%) of the WE patients were patch test positive. ACD was diagnosed in 29 out of 108 (26.9% of the WE patients and 40.8% of the PT positive patients) WE patients. ACD was also suspected in another 42 patients (39.8%), who had at least 1 positive PT result but the relevance of PT was hard to determine. In the remaining 37 patch test negative patients, 2 cases of food allergy were diagnosed by clinical findings and open food challenge test. 1 case of atopic dermatitis was diagnosed. 34 patients (31.5%) were diagnosed as unclassified endogenous eczema. Most of the WE patients with ACD (15/29, 51.7%) were ACD from widespread contact with hair dye during bathing. Most of the WE patients with hair dye ACD were male (male : female = 2.75). The total positive rates of PT in the WE group were no different to the control (65.7% vs 61.5%, p > 0.05, chi2-test). No significant difference was found for the positive rates of common contact allergens. The rate of atopy was no different between the 2 groups either (6.5% v 5.1%, p > 0.01, chi2-test). These results indicated that contact sensitization was very common in adult WE. Neglected widespread contact with contact allergens, especially hair dye, plays a very important role in our adult WE. The roles of age and sex in WE need to be studied further. PMID- 12581281 TI - Principles and methodology for identification of fragrance allergens in consumer products. AB - Fragrances contain several hundreds of different chemicals, a few major and many minor, which are responsible for the complexity of the odour. Fragrances are a major cause of allergic contact dermatitis. As a diagnostic tool, the current fragrance mix is very useful though not ideal. A 50-year-old woman presented with a pruriginous, erythematous eruption, characterized by papules, vesicles, exudation and crusting over the neck and chest. With the suspicion of fragrance allergy, patch testing was performed. Initially, the only positive reaction observed was with her own eau de toilette named Woman. The TRUE Test fragrance mix patch test was negative. Chemical fractionation of Woman perfume concentrate was combined with a sequenced patch testing procedure and with structure-activity relationship studies. Ingredients supplied by the manufacturer were also included in the study. Benzophenone-2, Lyral, alpha-hexyl cinnamic aldehyde and alpha damascone were found to be responsible for the patient's contact allergy to the commercial product. These substances contain chemical structural alerts giving them antigenic ability. The common use of new chemicals to manufacture fragrances, and the increased number of patients sensitive to them but with negative fragrance mix reactions, makes it necessary to identify new potential fragrance sensitizers in commercial products. PMID- 12581282 TI - Evaluation of metal allergies in patients with coronary stents. AB - In-stent restenosis is a complication which impairs the success of coronary stenting. Recently, it was supposed that a delayed hypersensitivity reaction to nickel and molybdenum might be one of the triggering factors in in-stent restenosis. We have analyzed the data collected in our centre with respect to this hypothesis. Altogether, 34 patients were investigated (24 male, 10 female). Patch tests were performed with the standard series of the German Contact Dermatitis Research Group and a metal series containing the metal components of 316 L stainless steel. A positive patch test reaction to nickel was observed in 4 (11.8%) patients. None of the patients showed sensitization to the other metals. Retrospective analysis was performed in 20 patients: 2 of these patients had a positive patch test reaction to nickel, one of whom had an in-stent restenosis, and the other not. Restenoses were predominantly observed in patients with negative patch test reactions to nickel (6/18 patients). Out of the patients who were investigated prospectively only one showed sensitization to nickel. Restenosis was observed in 2 patients: neither patient had nickel allergy. Although it still cannot be excluded that metal allergy may play a role in the restenosis process in coronary stenting, there is at present little evidence for it. PMID- 12581283 TI - Role of different valence states of chromium in the elicitation of allergic contact dermatitis. AB - Chromium, a known contact sensitizer, is present at high levels in certain detergent bars (40-50 p.p.m.) sold in India. A concern was whether chromium present in such detergent bars could sensitize users or their use lead to the elicitation of allergic contact dermatitis. Chromium present in detergent bars was analyzed for its valence state. It was found to be trivalent [Cr (III)] and not hexavalent [Cr (VI)]. An elicitation test on normal volunteers, as well as on volunteers who were sensitive to chromium, was carried out to determine the elicitation potential of chromium (III) in the bars. Of the 48 volunteers who completed the study, 30 did not show any positive response to either potassium dichromate, the positive control, or the detergent bar containing chromium (III). 18 volunteers who were identified as chromium-sensitive showed a positive response to potassium dichromate but not to the detergent bar containing trivalent chromium. This study clearly showed that the detergent bar with 40-50 p.p.m. of trivalent chromium did not elicit any allergic contact dermatitis even in individuals known to be chromium-sensitive. Hexavalent chromium (potassium dichromate) did however, elicit allergic contact dermatitis in these subjects. PMID- 12581284 TI - Positive patch test in hypersensitivity to lamotrigine. PMID- 12581285 TI - Biochemist's occupational allergic contact dermatitis from iodoacetamide and acrylamide. PMID- 12581286 TI - Contact allergy to topical corticosteroids 1995-2001. PMID- 12581287 TI - 'Canyoning hand': a new recreational hand dermatitis. PMID- 12581289 TI - Clinical and social changes in severely mentally ill individuals admitted to an outpatient psychosis team: an 18-month follow-up study. AB - AIM: The study investigated clinical and social changes during an 18-month follow up period in a group (n = 76) of schizophrenic outpatients admitted to a newly implemented outpatient psychosis team. Changes related to level of contact with the psychosis team were also examined as well as aspects of the content of the treatment interventions and work situation from a staff perspective. METHODS: Structured face-to-face interviews with the patients were performed at baseline and after 18 months. The Camberwell Assessment of Need instrument, the Lancashire Quality of Life Profile and the Interview Schedule for Social Interaction were used on both interview occasions along with Global Assessment of Functioning Scale and Brief Psychiatric Rating Scale. Thematic open-ended questions were used in staff interviews. RESULTS: Psychiatric symptoms, number of needs and number of met needs decreased, and perceived quality of life improved for the total sample during the follow-up period. Patients only in contact with a psychiatrist in the psychosis team improved more in symptoms and spent fewer days in hospital during follow-up time compared with those who had combined psychiatric and supportive contacts, and were also more satisfied with their medication. Patients with a combined contact deteriorated in psychosocial functioning compared with the group only in contact with a psychiatrist. Some of the elements in treatment interventions and work situation as well as hindrances in providing community based care adapted to the patients' needs were identified. CONCLUSIONS: Community based psychiatric services, to a larger extent, need to embrace evidence-based interventions and to perform regular, structured and comprehensive need assessments in order to ensure the effectiveness of interventions. Attention should be paid to staff motivation and education as well as to providing practical guidelines, supervision and support. PMID- 12581290 TI - Validation of the Parents' Postoperative Pain Measure in Finnish children aged 1 6 years. AB - The Canadian Parents' Postoperative Pain Measure (PPPM) is a behavioural pain measure for children. The purpose of this study was to test validity and reliability of this measure in 1-6-year-old Finnish children at home after minor day surgery. Children (n = 85) and their parents from four hospitals were included in the sample. The parents, mostly mothers (93%), filled in at home a questionnaire including PPPM during the day of operation, and the first and second postoperative days. Parents (n = 85) rated the presence or absence of behavioural changes from a checklist and also the child's worst pain during each day (using a Visual Analogue Scale, VAS). The response rate was 58. Principal component factor analysis, testing of correlation coefficient, cross-tabulations and Cronbach's alpha were used to test the instrument. The findings showed that construct validity of the measure was satisfactory. The PPPM was successful in discriminating between children who had no/low pain and children who had a clinically significant pain measured by VAS (convergence validity). Scores on the PPPM decreased from days 0 to 1 and from days 1 to 2 (predictive validity). Internal consistency of the measure and correlations with the pain scores on VAS were high on all days following surgery (equivalence). Sensitivity was satisfactory only when specificity was weaker. The findings of this study can be utilized in the development of children's postoperative pain assessment at home. However, further studies are needed to test the instrument in larger samples. PMID- 12581291 TI - Validity and reliability of the Nursing Minimum Data Set for the Netherlands (NMDSN). AB - The Nursing Minimum Data Set for the Netherlands (NMDSN) describes nursing care based on nursing phenomena, interventions and outcomes. The validity and reliability of its data collection has not been tested yet. PURPOSE: To report about the discriminative validity and the interrater reliability of the NMDSN. DESIGN: Data were collected in an intensive care ward, in a nursing home and in a residential home. The unit of measurement and analysis is the 'patient day'. The analysis for validity consisted of ridits calculations, and their graphical representations. Interrater reliability was measured by percentage agreement and Cohen's kappa. RESULTS: Graphs illustrate the differences on most nursing phenomena and interventions as expected beforehand. The percentage agreements for the residential home vary from 60.4 to 100%, and the kappa statistics from -0.09 to 0.85, indicating a poor to almost perfect interrater reliability. CONCLUSION: Intensive care patients and patients in the nursing home have more problems and need more nursing interventions compared with general hospital patients, while the patients in the residential home have lesser of both. This illustrates the discriminative validity of the NMDSN. The kappa values for various NMDSN variables are sufficient. A similar test in the general hospital is recommended. PMID- 12581292 TI - Maternal serum screening for Down syndrome--opinions on acceptance from Swedish women. AB - BACKGROUND: Different screening procedures are becoming an important part of health care. Information about screening and its consequences can be difficult to both impart and understand. This study examined women's theoretical acceptance of a new screening procedure, before its introduction. METHODS: A group of women (n = 823), who had made an informed choice about the form of foetal diagnosis they wished for, were also asked, in a questionnaire, about their opinion on the acceptability of serum screening for Down syndrome. The main purpose of the this was to ascertain their feelings when making a choice. RESULTS: Replies were received from 80.4%. The answers indicated that serum screening would be acceptable to 86% of the women who had chosen a second trimester ultrasound examination. Of the women who had chosen amniocentesis, 51% would consider the test acceptable as a first alternative. CONCLUSIONS: The ease with which the women were able to make their choice of prenatal diagnosis had bearing on their degree of acceptance of serum screening for Down syndrome. PMID- 12581293 TI - Breast cancer patients' experiences of lymphoedema. AB - The aim of this study was to explore employed women's experiences of light or moderate arm lymphoedema following breast cancer treatment in order to gain a deeper understanding of this phenomenon. Twelve women took part in a semistructured interview. A qualitative method with a phenomenological approach was applied to analyse data. In order to integrate the experiences in the everyday life of the women, a critical incident method was used. The findings indicate that there are many different practical and psychosocial problems related to arm lymphoedema. Three main themes were common to all the women. These themes were: (i) Attitudes from people in their surroundings, including reactions to the problem from other people and reactions from the women on the attitudes of other people. (ii) Discovery and understanding of oedema as a chronic disease and its treatment. (iii) Coping, including both problem-focused and emotion-focused strategies. The problems integrated in daily life were of low frequency but of considerable importance to the women. In conclusion, it is of great importance that health care professionals should be aware of and have knowledge about these problems. The women's needs for expressing their experiences of arm lymphoedema may be encouraged at the time of discovery and then regularly as long as the women seek care. Efforts may be made to strengthen the women's coping skills, eventually in a multidisciplinary approach. The interaction skills of health care professionals are probably of great importance in strengthening the resources of the women leading to a positive outcome. PMID- 12581294 TI - Patients' conceptions of coronary heart disease--a phenomenographic analysis. AB - Readjustment after an event of coronary heart disease (CHD) is defined to comprise cognitive, instrumental and affective components. The cognitive dimension refers to understanding of the disease. Twenty-three patients (<60 years) with CHD were interviewed about the nature of their disease and encouraged to use their own words. The study was conducted 1 year after the event of myocardial infarction (MI) and some patients had also been revascularized. The interviews were transcribed in extenso and analysed according to the phenomenographic approach. The main finding was the great variation of conceptions revealed. Six different conceptions were found concerning CHD. Some patients comprehended MI by involving (A) blood and vessels, (B) either blood or vessel or referred to (C) risk factors/symptoms. Angina pectoris was expressed as (A) insufficient heart capacity, (B) atherosclerosis/contracted vessel or as (C) symptoms. Several patients found it difficult to expand their answers and some expressed misconceptions about the course of events. Patients' pre-existing knowledge and their way of reasoning about central phenomena related to their disease should be taken into consideration in patient education and is also applicable in individual encounters with patients. PMID- 12581295 TI - Care dependency: testing the German version of the Care Dependency Scale in nursing homes and on geriatric wards. AB - The purpose of this study was the psychometric evaluation of the German version of the dependency scale in nursing homes and on geriatric wards. The 15-item scale was originally developed in the Netherlands for assessing the care dependency of demented and mentally handicapped patients. Data of 81 nursing home residents and of 115 geriatric patients were collected. Residents and patients were assessed several time by professionals and nonprofessionals. Reliability was determined by Cronbach's alpha, showing very good results with values of 0.94 and 0.98. Inter-rater and intrarater reliability show moderate to substantial Kappa values. Criterion validity was examined by comparing the data of the scale with the German nursing personal regulation and the German statutory insurance. The results show that the scores of the Care Dependency Scale correlate to the scores of the two classifications. In general, the German version of the scale can be recommended for use in nursing homes and on geriatric wards. PMID- 12581296 TI - Danish research-active clinical nurses overcome barriers in research utilization. AB - AIM: The aim of this study was to examine whether there was a difference between clinical nurses who were research-active, and clinical nurses who were nonresearch-active in utilization of research. A further aim was to identify the most significant barriers faced by a group of Danish clinical nurses in their use of research. BACKGROUND: Discrepancy between the improved quality of research results and the lack of implementing them was the starting point for a series of studies which showed the types of barriers clinical nurses found especially cumbersome when applying the research results of other researchers. This study investigates whether the clinical nurses' own engagement in research had any impact on their perception of research utilization. METHODOLOGY: The study had an exploratory and descriptive design. Seventy-nine Danish clinical nurses participated and semi-structured interviewing was used as the research method. FINDINGS: There was a statistically significant difference between the research active and nonresearch-active nurses on various variables. The study showed that, to a larger extent, research-active nurses used evidence-based knowledge and were generally more internationally orientated. Furthermore, two important barriers for research utilization were identified by all 79 clinical nurses included in the study, i.e. 90% of the nurses explained that the quantity of research results was overwhelming, and 75% of them found that they were unable to evaluate the quality of the research. CONCLUSIONS: Clinical nurses, who were research-active themselves, experienced more success in overcoming some of the barriers, which existed in applying research to practice. The research potential found amongst clinical nurses in Denmark needed to be further supported through training and guidance in research methodology, establishing introductory stipends and part time research positions. By doing so, some of the barriers affecting research utilization and the so-called theory-practice gap might be reduced. Further empirical studies, based on Giddens theory of sociological reflectivity, might see clinical nurses in a more serious light and simultaneously perceive them as producers of knowledge. PMID- 12581297 TI - Health promotion in the workplace: the perspective of unskilled workers in a hospital setting. AB - It is important to explore the potential for health promotion from the perspective of participants in a particular situation. This study focuses on experiences of well-being and the potential for health promotion among unskilled workers in the kitchen and laundry room in an acute care hospital in Iceland. These experiences are analysed in light of the material and structural factors that shape working conditions. The research method used was grounded theory. The data were formal and informal interviews, and participant observation. The findings revealed considerable differences between the two work groups. At both sites strain on the body, caused by excessive noise, heavy lifting and uncomfortable working arrangements, were identified. Similarly, the work was often experienced as monotonous and demanding. Human relations were identified as a key factor in ameliorating the conditions that led to experiences of strain by the workers at both work sites. This was however where the two work places differed. In the laundry room, the workers felt respected and acknowledged by their coworkers and managers. Belonging to the work group was important and they felt listened to by the supervisor. Very little cohesiveness was observed among the workers in the kitchen. The managers seemed to be absent from the floor, workers were divided into work groups that sometimes showed antagonism towards each other and few channels were available for complaints and suggestions for improvement. The workers in the laundry room, on the other hand, related positive experiences of work. They felt that steps were taken by managers to improve working conditions to the extent possible and participated in those attempts by bringing up suggestions, implementing change and developing their own ways of coping with the work. This study gives important directions for health promotion in the workplace, emphasizing human relations at work, rather than individual health behaviour away from the work setting. PMID- 12581298 TI - Ambiguity leads to uncertainty: ambiguous demands to blood donors. AB - Blood donors must respond to standard control questions and comply to certain demands before donating blood, to exclude possible recent contamination. This study investigates whether donors had adequate knowledge about 'the immunological window period', and whether the standard questionnaire in use was understandable and clear. It was found that nearly 40% of the donors had inadequate knowledge about 'the immunological window period' and that 10-40% of the donors would respond in various ways to different interpretations of the formulations of the questionnaire, indicating that the questionnaire was ambiguous. It was concluded that these factors could increase the risk of donating on wrong conditions. Demands and control questions posed to donors must be void of ambiguous formulations and take proper account of actual human sexual behaviours and life styles. Finally, it is suggested that blood banks must offer practical solutions (such as a 'fake donation') for temporarily rejected donors in order to respect their private integrity. PMID- 12581299 TI - Improvements in pain relief, handling time and pressure ulcers through internal audits of hip fracture patients. AB - The aim of this project was to improve the outcome of hip fracture patients by optimizing preoperative pain relief, diminishing the time from admission to operation and reducing the occurrence of pressure ulcers. A retrospective study of all medical records of hip fracture patients from the last 4 months in 1998 was compared with prospective registrations during the same period in 1999 and 2000 after the introduction of quality improvements. The number of patients who waited for more than 1 hour to get pain relief was almost halved after improvements. In 1998, close to half of the patients had to wait more than 24 hours for an operation. After attention was given to quality improvements, 36% of the patients in 1999 and 34% of the patients in 2000 had to wait more than 24 hours. In addition, 18% of the patients in 1999 and 24% of the patients in 2000 vs. 11% in 1998 were operated on within 12 had to wait more than 24 hours. Pressure ulcers were considerably reduced. In total, 19% of the patients in 1998, 8% in 1999 and 4.5% in 2000 had pressure ulcers at discharge from the hospital. The outcome for hip fracture patients was improved through attention to quality improvements with all staff involved and focused on these patients. PMID- 12581300 TI - Meaning of dependency on care as narrated by nurses. AB - This article is part of an ongoing study aimed at illuminating the meaning of dependency on care. The aim of this particular study is to illuminate the meaning of the phenomenon 'dependency on care' as narrated by nurses. Interviews with 20 nurses were conducted, tape-recorded and transcribed verbatim. A phenomenological hermeneutic approach was used to interpret the data. The meaning of dependency on care is revealed as laborious for both patients and nurses. The nurses' text discloses that for patients, dependency on care is a miserable loss of self determination and self-worth. Nurses appear to be moved by dependents' difficult life and want to do whatever they can for the dependent. Dependency on care is a burdensome responsibility for nurses. For nurses dependency on care is revealed as evoking feelings of guilt and insufficiency and of being constantly concerned and worried. It is to struggle to be patient and understanding, having the arduous task of putting oneself into the dependent patients' shoes in order to cope with the demanding task of balancing between helping and not helping too much. The interpretation discloses, however, that nurses distinguish between easy and trying dependency on care. The understanding indicates that when they meet 'good' patients, nurses reappraise the situation, transform it into something new and meaningful, that is dependency on care is disclosed as 'easy to take'. When coming across'difficult' patients dependency on care is disclosed as trying. It seems that within the unique nurse-patient relationship patients may participate in such a way as to facilitate or inhibit nurses in finding meaning in the immediate caregiving situation. Nurses' evaluations of what constitutes a 'good' or a 'difficult' patient, respectively, seem to play an important role in distinguishing between dependency on care that is easy and that which is trying. The result is viewed through an existential paradigm and there is also reflection about what nurses perceive as valuable qualities in patients which might influence the quality of care. PMID- 12581301 TI - Overexpression of Bax inhibitor suppresses the fungal elicitor-induced cell death in rice (Oryza sativa L) cells. AB - Treatment of suspension-cultured cells of rice (Oryza sativa L.) with cell wall extract of rice blast fungus (Magnaporthe grisea) elicits a rapid generation of H2O2, alkalinization of culture medium, and eventual cell death. To elucidate genes involved in these processes, we exploited SAGE (Serial Analysis of Gene Expression) technique for the molecular analysis of cell death in suspension cultured cells treated with the elicitor. Among the downregulated genes in the elicitor-treated cells, a BI-1 gene coding for Bax inhibitor was identified. Transgenic rice cells overexpressing Arabidopsis BI-1 gene showed sustainable cell survival when challenged with M. grisea elicitor. Thus, the plant Bax inhibitor plays a functional role in regulating cell death in the rice cell culture system. PMID- 12581302 TI - Touch modulates gravity sensing to regulate the growth of primary roots of Arabidopsis thaliana. AB - Plants must sense and respond to diverse stimuli to optimize the architecture of their root system for water and nutrient scavenging and anchorage. We have therefore analyzed how information from two of these stimuli, touch and gravity, are integrated to direct root growth. In Arabidopsis thaliana, touch stimulation provided by a glass barrier placed across the direction of growth caused the root to form a step-like growth habit with bends forming in the central and later the distal elongation zones. This response led to the main root axis growing parallel to, but not touching the obstacle, whilst the root cap maintained contact with the barrier. Removal of the graviperceptive columella cells of the root cap using laser ablation reduced the bending response of the distal elongation zone. Similarly, although the roots of the gravisensing impaired pgm1-1 mutant grew along the barrier at the same average angle as wild-type, this angle became more variable with time. These observations imply a constant gravitropic re-setting of the root tip response to touch stimulation from the barrier. In wild-type plants, transient touch stimulation of root cap cells, but not other regions of the root, inhibited both subsequent gravitropic growth and amyloplast sedimentation in the columella. Taken together, these results suggest that the cells of the root cap sense touch stimuli and their subsequent signaling acts on the columella cells to modulate their graviresponse. This interaction of touch and gravity signaling would then direct root growth to avoid obstacles in the soil while generally maintaining downward growth. PMID- 12581303 TI - Cell wall pectic (1-->4)-beta-d-galactan marks the acceleration of cell elongation in the Arabidopsis seedling root meristem. AB - Here we demonstrate that the pectic rhamnogalacturonan-I-associated LM5 (1-->4) beta-d-galactan epitope occurs in a restricted manner at the root surface of intact Arabidopsis seedlings. The root surface occurrence of (1-->4)-beta-d galactan marks the transition zone at or near the onset of rapid cell elongation and the epitope is similarly restricted in occurrence in epidermal, cortical and endodermal cell walls. The extent of surface (1-->4)-beta-d-galactan occurrence is reduced in response to genetic mutations (stp-1, ctr-1) and hormone applications that reduce root cell elongation. In contrast, the application of the arabinogalactan-protein (AGP) binding beta-glucosyl Yariv reagent (betaGlcY) that disrupts cell elongation results in the persistence of (1-->4)-beta-d galactan at the root surface and in epidermal, cortical and endodermal cell walls. This latter observation indicates that modulation of pectic (1-->4)-beta-d galactan may be an event downstream of AGP function during cell expansion in the Arabidopsis seedling root. PMID- 12581304 TI - Alteration of GCN5 levels in maize reveals dynamic responses to manipulating histone acetylation. AB - The role played by histone acetyltransferase (HAT), GCN5, in transcriptional co activation has been analysed in detail in yeast and mammals. Here, we present the cloning and expression pattern of Zmgcn5, the maize homologue. The enzymatic activity of the recombinant ZmGCN5 was analysed with histone and nucleosome substrates. In situ hybridisation of developing maize kernels using Zmgcn5 as probe shows that the transcript is concentrated in rapidly dividing cells. To investigate the role of ZmGCN5 in the transcription of specific plant genes, direct protein-protein interactions were tested. A cDNA clone encoding a putative interacting partner in GCN5-adapter complexes, ZmADA2, was isolated and the interaction between ZmGCN5 and ZmADA2 was confirmed by a GST-spin down experiment. Co-immunoprecipitation of the plant transcriptional activator Opaque 2 and ZmADA2 in nuclear extracts suggests ADA2/GCN5-containing complexes to mediate transcriptional activation by binding of this bZIP factor. For a more general analysis of the effects of histone acetylation on plant gene expression, 2500 ESTs spotted on filters were hybridised with cDNA probes derived either from maize cell lines treated with Trichostatin A (TSA), or from a transgenic line expressing the ZmGCN5 antisense transcript. Several sequences showing marked changes in abundance were confirmed by RNA blot analysis. Inhibition of histone deacetylation with TSA is accompanied by a decrease in the abundance of ZmGCN5 acetylase protein, but by increases in mRNAs for histones H2A, H2B, H3 and H4. The elevated histone mRNA levels were not reflected in increasing histone protein concentrations, suggesting hyperacetylated histones arising from TSA treatment may be preferentially degraded and substituted by de novo synthesised histones. The ZmGCN5 antisense material showed suppression of the endogenous ZmGCN5 transcript and the profiling analysis revealed increased mRNA levels for H2A, H2B and H4. Furthermore, in the antisense line, a reduction in the amount of the RPD3 type HD1B-I histone deacetylase protein was observed. A model for linked regulation of histone acetylation and histone mRNA transcription is discussed. PMID- 12581305 TI - Histone modifications in Arabidopsis- high methylation of H3 lysine 9 is dispensable for constitutive heterochromatin. AB - N-terminal modifications of nucleosomal core histones are involved in gene regulation, DNA repair and recombination as well as in chromatin modeling. The degree of individual histone modifications may vary between specific chromatin domains and throughout the cell cycle. We have studied the nuclear patterns of histone H3 and H4 acetylation and of H3 methylation in Arabidopsis. A replication linked increase of acetylation only occurred at H4 lysine 16 (not for lysines 5 and 12) and at H3 lysine 18. The last was not observed in other plants. Strong methylation at H3 lysine 4 was restricted to euchromatin, while strong methylation at H3 lysine 9 occurred preferentially in heterochromatic chromocenters of Arabidopsis nuclei. Chromocenter appearance, DNA methylation and histone modification patterns were similar in nuclei of wild-type and kryptonite mutant (which lacks H3 lysine 9-specific histone methyltransferase), except that methylation at H3 lysine 9 in heterochromatic chromocenters was reduced to the same low level as in euchromatin. Thus, a high level of H3methylK9 is apparently not necessary to maintain chromocenter structure and does not prevent methylation of H3 lysine 4 within Arabidopsis chromocenters. PMID- 12581306 TI - A role for HvGAMYB in anther development. AB - HvGAMYB is a transcription factor that was first identified in barley aleurone cells and shown to be upregulated by gibberellin (GA). Using RNA and immunoblot analysis we have shown HvGAMYB is also strongly expressed in barley anthers. Transgenic barley expressing a HvGAMYB:GFP fusion gene have been created and GAMYB expression in anthers analysed. GFP expression was clearly visible during early anther development in the nuclei of the epidermis, endothecium, middle layer and tapetum. Expression in the epidermis and endothecium persists until just prior to anther dehiscence, expression in the other two cell layers is visible until they are compressed and broken down as the microspores develop. Further evidence of a role for HvGAMYB in anther development was provided by the creation of transgenic barley over-expressing the HvGAMYB gene. Associated with the increase in HvGAMYB levels was a progressive decrease in anther size, particularly a decrease in anther length. Anthers also became increasingly lighter in colour. Anthers with fourfold more HvGAMYB protein than non-transgenic controls failed to dehisce and were male sterile, anthers with approximately three to fourfold endogenous GAMYB protein levels were smaller and paler but still shed normally. To investigate the hormonal regulation of HvGAMYB expression in anthers, HvGAMYB and SLN1 protein levels in anthers were analysed following application of GA3. As in cereal aleurone, HvGAMYB levels were found to increase and SLN1 levels decrease following GA3 application suggesting a similar GA signalling pathway to that in aleurone exists in anthers. PMID- 12581307 TI - A novel ER-derived compartment, the ER body, selectively accumulates a beta glucosidase with an ER-retention signal in Arabidopsis. AB - The ER body is a novel compartment that is derived from endoplasmic reticulum (ER) in Arabidopsis. In contrast to whole seedlings which have a wide distribution of the ER bodies, rosette leaves have no ER bodies. Recently, we reported that wound stress induces the formation of many ER bodies in rosette leaves, suggesting that the ER body plays a role in the defense system of plants. ER bodies were visualized in transgenic plants (GFP-h) expressing green fluorescent protein (GFP) with an ER-retention signal, HDEL. These were concentrated in a 1000-g pellet (P1) of GFP-h plants. We isolated an Arabidopsis mutant, nai1, in which fluorescent ER bodies were hardly detected in whole plants. We found that a 65-kDa protein was specifically accumulated in the P1 fraction of GFP-h plants, but not in the P1 fraction of nai1 plants. N-terminal peptide sequencing revealed that the 65-kDa protein was a beta-glucosidase, PYK10, with an ER-retention signal, KDEL. Immunocytochemistry showed that PYK10 was localized in the ER bodies. Compared with the accumulation of GFP-HDEL, which was associated with both cisternal ER and ER bodies, the accumulation of PYK10 was much more specific to ER bodies. PYK10 was one of the major proteins in cotyledons, hypocotyls and roots of Arabidopsis seedlings, while PYK10 was not detected in rosette leaves that have no ER bodies. These findings indicated that PYK10 is the main component of ER bodies. It is possible that PYK10 produces defense compounds when plants are damaged by insects or wounding. PMID- 12581308 TI - ADP-glucose pyrophosphorylase from potato tuber: site-directed mutagenesis of homologous aspartic acid residues in the small and large subunits. AB - Asp142 in the homotetrameric ADP-glucose pyrophosphorylase (ADP-Glc PPase) enzyme from Escherichia coli was demonstrated to be involved in catalysis of this enzyme [Frueauf, J.B., Ballicora, M.A. and Preiss J. (2001) J. Biol. Chem., 276, 46319 46325]. The residue is highly conserved throughout the family of ADP-Glc PPases, as well as throughout the super-family of sugar-nucleotide pyrophosphorylases. In the heterotetrameric ADP-Glc PPase from potato (Solanum tuberosum L.) tuber, the homologous residue is present in both the small (Asp145) and the large (Asp160) subunits. It has been proposed that the small subunit of plant ADP-Glc PPases is catalytic, while the large subunit is modulatory; however, no catalytic residues have been identified. To investigate the function of these conserved Asp residues in the ADP-Glc PPase from potato tuber, we used site-directed mutagenesis to introduce either an Asn or a Glu. Kinetic analysis in the direction of synthesis or pyrophosphorolysis of ADP-Glc showed a significant decrease (more than four orders of magnitude) in the specific activity of the SD145NLwt, SD145NLD160N, and SD145NLD160E mutants, while the effect was smaller (approximately two orders of magnitude) with the SD145ELwt, SD145ELD160N, and SD145ELD160E mutants. By contrast, mutation of the large subunit alone did not affect the specific activity but did alter the apparent affinity for the activator 3 phosphoglycerate, showing two types of apparent roles for this residue in the different subunits. These results show that mutation of Asp160 of the large subunit does not affect catalysis, thus the large subunit is not catalytic, and that the negative charge of Asp145 in the small subunit is necessary for enzyme catalysis. PMID- 12581309 TI - The OsTB1 gene negatively regulates lateral branching in rice. AB - Although the shoot apical meristem (SAM) is ultimately responsible for post embryonic development in higher plants, lateral meristems also play an important role in determining the final morphology of the above-ground part. Axillary buds developing at the axils of leaves produce additional shoot systems, lateral branches. The rice TB1 gene (OsTB1) was first identified based on its sequence similarity with maize TEOSINTE BRANCHED 1 (TB1), which is involved in lateral branching in maize. Both genes encode putative transcription factors carrying a basic helix-loop-helix type of DNA-binding motif, named the TCP domain. The genetic locus of OsTB1 suggested that OsTB1 is a real counterpart of maize TB1. Transgenic rice plants overexpressing OsTB1 exhibited markedly reduced lateral branching without the propagation of axillary buds being affected. We also demonstrated that a rice strain carrying a classical morphological marker mutation, fine culm 1 (fc1), contain the loss-of-function mutation of OsTB1 and exhibits enhanced lateral branching. Expression of OsTB1, as examined with a putative promoter-glucuronidase (GUS) gene fusion, was observed throughout the axillary bud, as well as the basal part of the shoot apical meristem, vascular tissues in the pith and the lamina joint. Taking these data together, we concluded that OsTB1 functions as a negative regulator for lateral branching in rice, presumably through expression in axillary buds. PMID- 12581310 TI - Mutations in a gene for plastid ribosomal protein S6-like protein reveal a novel developmental process required for the correct organization of lateral root meristem in Arabidopsis. AB - Arabidopsis rfc3 mutants have previously been isolated with an altered fatty acid composition of membrane lipids. In this study, rfc3 was found to have a sucrose conditional defect in the patterning of distal elements in the lateral root meristem. By utilizing this feature, a sucrose-sensitive process important for lateral root development was localized to the growing portion of rfc3 primary root. Because lateral root formation occurs at a later stage, this finding suggests the existence of an RFC3-dependent, non-primordium autonomous signal playing a role in the organization of lateral root meristem. Map-based cloning of RFC3 gene revealed that it encodes a plastid-localized ribosomal protein S6-like protein and provides a potential link between control of plastid gene expression and LR development. PMID- 12581312 TI - An abscisic acid-sensitive checkpoint in lateral root development of Arabidopsis. AB - Lateral root (LR) formation displays considerable plasticity in response to developmental and environmental signals. The mechanism whereby plants incorporate diverse regulatory signals into the developmental programme of LRs remains to be elucidated. Current concepts of lateral root regulation focus on the role of auxin. In this study, we show that another plant hormone, abscisic acid (ABA), also plays a critical role in the regulation of this post-embryonic developmental event. In the presence of exogenous ABA, LR development is inhibited. This occurs at a specific developmental stage, i.e. immediately after the emergence of the LR primordium (LRP) from the parent root and prior to the activation of the LR meristem, and is reversible. Interestingly, this inhibition requires 10-fold less ABA than the inhibition of seed germination and is only slightly reduced in characterised abi mutants, suggesting that it may involve novel ABA signalling mechanisms. We also present several lines of evidence to support the conclusion that the ABA-induced lateral root inhibition is mediated by an auxin-independent pathway. First, the inhibition could not be rescued by either exogenous auxin application or elevated auxin synthesis. Secondly, a mutation in the ALF3 gene, which is believed to encode an important component in the auxin-dependent regulatory pathway for the post-emergence LR development, does not affect the sensitivity of LRs to ABA. Thirdly, ABA and the alf3-1 mutation do not act at the same developmental point. To summarise, these results demonstrate a novel ABA sensitive, auxin-independent checkpoint for lateral root development in Arabidopsis at the post-emergence stage. In addition, we also present data indicating that regulation of this developmental checkpoint may require novel ABA signalling mechanisms and that ABA suppresses auxin response in the LRPs. PMID- 12581311 TI - Structure and expression of the rice class-I type histone deacetylase genes OsHDAC1-3: OsHDAC1 overexpression in transgenic plants leads to increased growth rate and altered architecture. AB - Histone deacetylases (HDACs) modulate chromatin structure and transcription. HDACs have been studied as negative regulators in eukaryotic transcription. We isolated the rice OsHDAC1-3 genes for class I-type histone deacetylases, which are related to the RPD3 family. The OsHDAC1 gene encoded a protein of approximately 57 kDa that shared 73.5, 72.7, 79.9, and 57.1% amino acid sequence identity with the OsHDAC2, OsHDAC3, maize RPD3, and human HDAC1 proteins, respectively. Genomic structures and Southern blot analyses revealed that OsHDAC1 3 contained seven, six, and seven exons, respectively, and constituted a class I type family in the rice genome. OsHDAC1 was expressed at similar levels in the leaves, roots, and callus cells, whereas OsHDAC2 and 3 were expressed in the roots and callus cells, but not in the leaves, exhibiting distinct tissue specificity. To explore the role of histone deacetylases in transgenic plants, we inserted the OsHDAC1 cDNA fragment into the expression vector Ai::OsHDAC1 under the control of the ABA-inducible promoter Ai, and transformed the construct into rice. Levels of mRNA, protein, and HDAC activity were significantly increased in Ai::OsHDAC1 callus cells. The amount of tetra-acetylated H4 in the transgenic cells was greatly reduced, and the reduction was abolished upon treatment with trichostatin A. These results demonstrate that OsHDAC1 overexpression in transgenic cells both yields enzymatically active HDAC complexes and induces changes in histone acetylation in vivo. The overexpression leads to a range of novel phenotypes, involving increased growth rate and altered plant architecture, suggesting that OsHDAC1 functions in the genome-wide programming of gene expression. PMID- 12581313 TI - Selection by phage display of a variant mustard trypsin inhibitor toxic against aphids. AB - The mustard trypsin inhibitor, MTI-2, is a potent inhibitor of trypsin with no activity towards chymotrypsin. MTI-2 is toxic for lepidopteran insects, but has low activity against aphids. In an attempt to improve the activity of the inhibitor towards aphids, a library of inhibitor variants was constructed and cloned into the pRlac3 phagemid vector. The library of 9.3 x 107 independent colonies was created by randomisation of a stretch of five consecutive codons in the reactive site. Repeated selection rounds against bovine trypsin and chymotrypsin allowed the identification of novel, MTI-2 derived, antitrypsin and antichymotrypsin inhibitors. Chy8, the selected variant with highest affinity for bovine chymotrypsin (Ki = 32 nm versus >1000 nm for the wild-type) represents the strongest known recombinant chymotrypsin inhibitor of the MTI-2 family. It is highly toxic to nymphs of the aphid Acyrthosiphon pisum, and moderately toxic to nymphs of Aphis gossypii and Myzus persicae. The LC50 of 73 microg ml-1 towards A. pisum is the lowest value known among chymotrypsin inhibitors. The aphicidal activity of Chy8 was improved eightfold compared to the wild-type inhibitor. This demonstrates, for the first time, that bovine chymotrypsin provides a useful template to select engineered proteins highly toxic against these aphids. The selected gene will allow the development of transgenic crops that are protected against sucking insect pests. PMID- 12581314 TI - The tomato mutant spr1 is defective in systemin perception and the production of a systemic wound signal for defense gene expression. AB - Wound-induced systemic expression of defensive proteinase inhibitor (PI) genes in tomato plants requires the action of systemin and its precursor protein prosystemin. Although it is well established that systemin induces PI expression through the octadecanoid pathway for jasmonic acid (JA) biosynthesis, relatively little is known about how systemin and JA interact to promote long-distance signaling between damaged and undamaged leaves. Here, this question was addressed by characterizing a systemin-insensitive mutant (spr1) that was previously identified as a suppressor of prosystemin-mediated responses. In contrast to JA biosynthetic or JA signaling mutants that lack both local and systemic PI expression in response to wounding, spr1 plants were deficient mainly in the systemic response. Consistent with this phenotype, spr1 plants exhibited normal PI induction in response to oligosaccharide signals that are thought to play a role in the local wound response. Moreover, spr1 abolished JA accumulation in response to exogenous systemin, and reduced JA accumulation in wounded leaves to approximately 57% of wild-type (WT) levels. Analysis of reciprocal grafts between spr1 and WT plants showed that spr1 impedes systemic PI expression by blocking the production of the long-distance wound signal in damaged leaves, rather than inhibiting the recognition of that signal in systemic undamaged leaves. These experiments suggest that Spr1 is involved in a signaling step that couples systemin perception to activation of the octadecanoid pathway, and that systemin acts at or near the site of wounding (i.e. in rootstock tissues) to increase JA synthesis to a level that is required for the systemic response. It was also demonstrated that spr1 plants are not affected in the local or systemic expression of a subset of rapidly induced wound-response genes, indicating the existence of a systemin-independent pathway for wound signaling. PMID- 12581316 TI - An Arabidopsis mutant defective in UV-B light-mediated responses. AB - In plants, low fluences of ultraviolet-B radiation (UV-B; 280-320 nm) trigger the activation of protection mechanisms and various photomorphogenic responses. We established a novel screen to isolate Arabidopsis T-DNA mutants (uli, UV-B light insensitive) with reduced sensitivity towards UV-B light-mediated inhibition of hypocotyl growth. One of the mutants, uli3, shows also a reduced sensitivity of UV-B-induced gene expression events and was, therefore, chosen for further investigation. The ULI3 gene encodes an 80-kDa protein with potential domains for heme- and diacylglycerol-binding. In transiently transfected protoplasts, the ULI3:GFP fusion protein is localised in the cytoplasm but also adjacent to membranes. In etiolated Arabidopsis seedlings irradiated by UV-B, ULI3 mRNA expression is strongly up-regulated. This is caused by elevated transcription as demonstrated using stable transformants where a GUS-reporter was driven by the ULI3-promoter. ULI3 is preferentially expressed in the outer cell layers in leaves, stems and flowers, but not in roots. There is evidence that ULI3 represents a specific component involved in UV-B-mediated signal transduction in higher plants. PMID- 12581315 TI - Allene oxide cyclase dependence of the wound response and vascular bundle specific generation of jasmonates in tomato - amplification in wound signalling. AB - The allene oxide cyclase (AOC)-catalyzed step in jasmonate (JA) biosynthesis is important in the wound response of tomato. As shown by treatments with systemin and its inactive analog, and by analysis of 35S::prosysteminsense and 35S::prosysteminantisense plants, the AOC seems to be activated by systemin (and JA) leading to elevated formation of JA. Data are presented on the local wound response following activation of AOC and generation of JA, both in vascular bundles. The tissue-specific occurrence of AOC protein and generation of JA is kept upon wounding or other stresses, but is compromised in 35S::AOCsense plants, whereas 35S::AOCantisense plants exhibited residual AOC expression, a less than 10% rise in JA, and no detectable expression of wound response genes. The (i). activation of systemin-dependent AOC and JA biosynthesis occurring only upon substrate generation, (ii). the tissue-specific occurrence of AOC in vascular bundles, where the prosystemin gene is expressed, and (iii). the tissue-specific generation of JA suggest an amplification in the wound response of tomato leaves allowing local and rapid defense responses. PMID- 12581319 TI - Human herpesvirus 8: Is it time for routine surveillance in pediatric solid organ transplant recipients to prevent the development of Kaposi's sarcoma? PMID- 12581320 TI - Cytotoxic T lymphocyte monitoring in steroid-free immunosuppression. PMID- 12581321 TI - Indications and outcome of treatment of hyperlipidemia in pediatric allograft recipients. PMID- 12581322 TI - Human herpesvirus-6 and -7 infections in transplantation. AB - Human herpesvirus-6 (HHV-6) and human herpesvirus-7 (HHV-7) are ubiquitous in the human population and cause exanthem subitum, a benign disease seen in infancy. The viruses remain latent in the body after primary infection, and reactivate in immunocompromised patients. HHV-6 infection occurs in nearly 50% of all bone marrow and in 20-30% of solid-organ transplant recipients, 2-3 weeks following the procedure. It has been suggested that the viral infection and activation result in clinical symptoms, including fever, skin rash, pneumonia, bone marrow suppression, encephalitis, and rejection. In order to understand the viral infection in greater detail, several studies investigating the route of viral transmission and diagnostic procedures have been carried out. In contrast to studies of HHV-6 infection in organ-transplant recipients, the number of studies examining HHV-7 infection in these patients is limited. According to several recent studies, HHV-7 may act as a cofactor for cytomegalovirus disease in organ transplant recipients. PMID- 12581323 TI - Glomerular filtration rate as a putative 'surrogate end-point' for renal transplant clinical trials in children. AB - Only with prospective randomized controlled trials is it possible to evaluate the several immunosuppressive regimens available to renal allograft recipients. Commonly used surrogate markers of clinical outcome, such as patient and graft survival, are constantly improving. Current immunosuppressive protocols have improved 1-yr graft survival to over 90%. The small differences in graft survival among the various immunosuppressive regimes require large patient cohorts in order to establish statistical significance. Such studies are often difficult to conduct in a timely manner, particularly in children. This necessitates the search for better surrogate markers sensitive enough to detect differences in smaller cohorts and in a shorter period of time. While the degree of fibrosis in transplant biopsies might well predict long-term graft survival, protocol biopsies are expensive, invasive, and unpopular among clinicians. In native kidneys, glomerular filtration rate (GFR) closely correlates with disease progression and interstitial fibrosis and appears to be well positioned as a less invasive surrogate marker for long-term outcome. Nonetheless, the ideal marker for GFR remains obscure. Serum creatinine has several major drawbacks, making it a poor predictor of GFR. This review discusses the several methods used to estimate or measure GFR with emphasis on 125I-iothalamate clearance and serum cystatin C (cys-C). Of all the serum markers, cys-C is the most reliable and the most promising. However, cys-C and other endogenous markers cannot replace the diagnostic sensitivity and reliability of radiolabeled markers of GFR such as 125I-iothalamate in renal transplant clinical trials. Unfortunately, clearance of most radiolabeled markers of GFR including 125I-iothalamate remain costly and time consuming. PMID- 12581325 TI - Megakaryocyte chimerism after allogeneic stem cell transplantation in children. AB - The aim of this study was to investigate the extent and the clinical implications of mixed chimerism in megakaryocytes after stem cell transplantation (SCT). Polymerase chain reaction analyzing allele length polymorphisms was used to determine the origin of immunomagnetically isolated megakaryocytes and leukocyte subpopulations after SCT in 13 children. Eleven were unselected consecutive cases while two were included due to known leukocyte mixed chimerism. Recipient DNA was detected in the megakaryocytes in six out of the 11 cases at levels between 1 and 100%. Coinciding mixed chimerism in the leukocyte populations was detected in two of the 11 cases. Of the two selected cases with known leukocyte mixed chimerism, two boys with aplastic anemia and Wiskott-Aldrich syndrome had 1-5 and 70% recipient megakaryocytes, respectively. Although the four relapses or deaths, within the 13 months of observation, were restricted to patients with multilineage or isolated megakaryocyte (n = 1) mixed chimerism, it was not possible to link any other apparent clinical problems, except a prolonged thrombocytopenia in one case, to the mixed chimerism in this limited study group. PMID- 12581324 TI - Neoral monitoring 2 hours post-dose and the pediatric transplant patient. AB - Cyclosporin A therapy has evolved greatly over the past 25 years of clinical experience. Sophisticated studies of CsA pharmacokinetics and pharmacodynamics have led to a better understanding of the relationship between dose response and biological effect. It has become apparent that achieving target drug exposure is necessary for optimal clinical outcomes. Monitoring dose response has become a key aspect of immunosuppressive management. This review presents the information available supporting cyclosporin drug concentration drawn two hours post dose (C 2) in children who have been transplanted as the best single indicator of CsA exposure. Further studies evaluating the clinical benefit of achieving C-2 targets in children are indicated. PMID- 12581326 TI - Atorvastatin treatment for hyperlipidemia in pediatric renal transplant recipients. AB - The objective of this prospective study was to determine the prevalence of hyperlipidemia in our pediatric renal transplant patients and to treat those with persistently elevated cholesterol and/or low-density lipoprotein (LDL) levels. All patients with a functioning renal allograft for greater than 6 months were studied (n = 18). Patients with cholesterol and/or LDL levels greater than the 95th percentile (n = 9) were commenced on an HMG-CoA reductase inhibitor, Atorvastatin and monitoring was performed for efficacy and adverse effects. Total serum cholesterol was elevated in 11 of 18 (61%) and triglyceride (TG) was elevated in 12 of 18 (67%) patients. Atorvastatin treatment was effective with a mean percentage reduction of total cholesterol of 41 +/- 10% (p < 0.01 vs. before treatment), LDL 57 +/- 7% (p < 0.01 vs. before treatment) and TG 44 +/-25% (p = 0.05 vs. before treatment). No adverse effects on allograft function or cyclosporin levels were experienced. Hyperlipidemia is a common problem and Atorvastatin is a safe and effective treatment in pediatric renal transplant recipients. PMID- 12581327 TI - Cardiac troponin I: a marker of acute heart rejection in infant and child heart recipients? AB - Acute rejection of the donor heart is a major cause of mortality in infant heart transplant recipients. The early diagnosis of acute cardiac rejection (ACR) is crucial. Non-invasive methods have shown poor sensitivity in detecting rejection when compared to endomyocardial biopsies (EMB). We assessed troponin I as a new marker to diagnose cardiac rejection. Serum cardiac troponin I (cTNI) levels were retrospectively analysed in 25 heart transplant patients (ages, 2 wk to 13 yr; mean age, 3 months) presenting 36 acute rejections. In early post-operative rejection and initially elevated cTNI levels, rejection was associated with a second increase of serum cTNI concentrations in 21% of the patients (p = 0.15). If cTNI levels were in normal range before ACR an elevation was monitored in 59% of the rejection periods (p < 0.05). In 25% of the cases (n = 9) cTNI levels remained in normal range during the rejection episode (<0.6 ng/mL), in 22% (n = 8) cTNI levels did not exceed pathological values from 0.6 to 1.5 ng/mL and in 53% (n = 19) the measured levels went beyond 1.5 ng/mL. Maximum concentrations of cTNI were measured mostly 12 d from the moment rejection was suspected (day 1) in patients (median day 3). However, cTNI levels were elevated for 2-43 d after ACR was diagnosed (median 10 d). Twenty per cent of the patients with grade 3 rejection (ISHLT) and 75% of the patients with grade 4 rejection had a corresponding elevated cTNI level (p = 0.013). No false-positive elevations of cTNI were documented. The present data demonstrate that cTNI is a not a sensitive but a specific marker of ACR in children. PMID- 12581328 TI - Serial detection of Epstein-Barr virus DNA in sera and peripheral blood leukocyte samples of pediatric renal allograft recipients with persistent mononucleosis like symptoms defines patients at risk to develop post-transplant lymphoproliferative disease. AB - We tested blood samples of 25 pediatric renal transplant recipients for Epstein Barr virus (EBV) DNA load by quantitative polymerase chain reaction (PCR). Eleven of these transplant recipients showed clinical persistent mononucleosis-like symptoms years after transplantation (Tx). A quantitation of EBV DNA by PCR in peripheral blood lymphocyte (PBL) and serum samples revealed variable EBV DNA titers. The majority of EBV PCR results in samples of the 14 asymptomatic transplant recipients was repeatedly below detection limit. In contrast, patients with mononucleosis-like symptoms showed persistent EBV genome titers over a period of 6 months, ranging from 75 to 18 750 copies/10 000 PBL and from 680 to 335 000 copies/mL serum, respectively. One child suffering from this mononucleosis-like condition developed an EBV-associated Burkitt-like lymphoma 29 months after Tx. Whereas clinical and histological investigations did not indicate a post-transplant lymphoproliferative disorder (PTLD) until tumor detection, EBV titers in PBL and serum had been high for at least 8 months. We propose that pediatric transplant recipients who show both, recurrent mononucleosis-like symptoms and a sustained high EBV genome load, are at increased risk for severe EBV-related post-transplant complications. PMID- 12581329 TI - Increased expression of cytotoxic effector molecules: different interpretations for steroid-based and steroid-free immunosuppression. AB - Cytotoxic T lymphocyte (CTL) effector molecules have been studied as markers of acute rejection in renal allograft recipients on steroid-based immunosuppression. We hypothesized that basal CTL gene expression may vary with time post transplantation as well as with different immunosuppression protocols (steroid based or steroid-free). Variations in CTL gene expression may thus impact on the ability to predict acute allograft rejection. We used the non-invasive method of quantitative competitive-reverse transcription-polymerase chain reaction (QC-RT PCR) to quantify the amounts of CTL effector molecules (granulysin, GL; perforin, P; granzyme B, GB) in serial peripheral blood lymphocyte (PBL) samples from steroid-free and steroid-based adult and pediatric renal allograft recipients. Patients on both protocols were clinically monitored by protocol biopsies at 1, 3, 6, and 12 months post-transplantation and for graft function at 1 yr post transplantation in a separate clinical study. Steroid-free patients with stable graft function showed an increase in GL, P, and GB gene expression over time post transplantation with the increase being seen largely by the first post-transplant month. A further increase in GL expression was noted at the end of the first post transplant year in the absence of acute rejection, whereas GB and P levels were unchanged. At comparative time-points post-transplantation, CTL genes were found to be higher in steroid-free patients with stable graft function, compared to steroid-based recipients with either clinically stable graft function or acute rejection. This study suggests that levels of CTL gene expression, although important in a steroid-based regimen to monitor the risk of acute rejection, may not be similarly applied in patients on steroid-free immunosuppression. The early increase in levels seen in steroid-free patients appears to correlate with the total absence of steroids. As steroid-free patients seem to have a lower incidence of acute rejection and better long-term graft function at 1 yr, the early increase in CTL genes in the absence of acute rejection may suggest an early adaptive immune activation response, promoting early graft acceptance in this protocol. PMID- 12581330 TI - Pediatric cardiac transplant: results using a steroid-free maintenance regimen. AB - We report on survival, rejection, lymphoma and renal function following cardiac transplant using a steroid-free maintenance immunosuppressive regimen. We have performed 73 cardiac transplants in 71 children under 16 yr of age in the last 12 yr. There were eight perioperative and four late deaths giving actuarial survival of 88, 88, 85 and 70% at 1, 2, 5 and 10 yr, respectively. A total of 11 (15.3%) children had one episode of rejection (grade 3) in the first 6 months; one died and one was re-transplanted because of rejection. There was only one episode of late rejection (8 yr post-transplant) because of low drug levels in a patient with lymphoma and sepsis. This patient did not survive. Three other children (5.6%) also developed lymphoma and recovered but one died subsequently of graft failure. Four children have developed severe renal failure (glomerular filtration rate GFR <30 mL/min/m2). Two have not survived and one is expected to commence dialysis soon. The remainder have mild to moderate renal impairment. We report excellent survival and low rejection rates without use of long-term steroids. However the doses of cyclosporin used have had a significant effect on renal function in many cases. PMID- 12581331 TI - The pediatric risk of mortality score in infants and children with fulminant liver failure. AB - The pediatric risk of mortality (PRISM) score as a severity scoring system has never been assessed in infants and children with fulminant liver failure (FLF). A retrospective case study of 109 infants and children admitted in a 22-bed pediatric and neonatal intensive care unit of a tertiary university hospital, National Referral Center for Pediatric Liver Transplantation, from March 1986 to August 1997 was carried out. PRISM score was not significantly different within etiologic FLF categories, or between infants and children. However, PRISM score (mean +/- SD) showed significant difference (p = 0.001) between the 27 patients who spontaneously recovered with supportive care (8.8 +/- 5.0) and 82 patients who underwent emergency liver transplantation (ELT) or those who died before (14.9 +/- 7.7). PRISM score-based probability of mortality was underestimated when compared with observed mortality. A death probability higher than 20% had a 24% sensitivity and 95% specificity for severe outcome. Reciever operating characteristic curve for PRISM score showed elevated discriminative power (Az = 0.91) for discerning children with severe outcome from those who spontaneously recovered with supportive care. A PRISM score more than 10 showed an odds ratio of 2.69 for predicting severe outcome (95% CI: 1.11-6.55; p = 0.038). In conclusion, the PRISM score is an accurate means of severity assessment in pediatric FLF. However, PRISM score-based mortality was of low predictive value. PMID- 12581332 TI - Renal transplantation: long-term adaptation and the children's own reflections. AB - Twenty children, 9-19 yr of age, having undergone renal transplantation, were interviewed (according to a semi-structured inquiry manual) with the aim of investigating their emotional and psychosocial adaptation. The children reported about a rather unaffected life considering the circumstances. Few of them described a negative identity in the form of feeling sick or different, and their experienced limitations appeared to be moderate. Anxiety about the future was more salient, but nevertheless the children seemed to have found a good way of living with this anxiety. When comments were made on the relationship to the donor, they were of a positive character; the relationship had been improved by the donation. Only one child - who had received a kidney from a deceased person - described fantasies of the alien kidney of a markedly unpleasant character. The psychological adaptation of the children frequently seemed to imply the use of potentially dysfunctional defense mechanisms like denial and avoidance. PMID- 12581333 TI - Clostridium difficile colitis associated with inflammatory pseudotumor in a liver transplant recipient. AB - The aim of this report is to describe a rare complication of clostridium difficile (CD) disease, the occurrence of an inflammatory pseudotumor that caused intestinal obstruction in a liver transplant recipient. A 9-month-old girl underwent liver transplantation for biliary atresia. She was given tacrolimus as primary immunosuppressive therapy. Three months after liver transplantation, she presented with febrile protracted bloody diarrhea and failure to thrive. A diagnosis of post-transplant lymphoproliferative disease associated with Epstein Barr virus infection was initially made on histological examination of duodenal biopsies. Tacrolimus was discontinued. Despite treatment with anti-CD20 monoclonal antibodies the child's condition deteriorated and she presented with intestinal occlusion. A mass at the ascending colon was seen on the computed tomography scan mimicking lymphoma and the child underwent surgical laparotomy. Histological examination of the mass showed typical pathological lesions of inflammatory pseudotumor and CD pseudomembranous colitis. Diagnosis of CD disease was confirmed upon the identification of CD toxin A in stools. She was successfully treated by metronidazole and gamma-globulin perfusions. Delayed diagnosis and anti-CD20 monoclonal antibodies therapy (associated with hypogammaglobulinemia) possibly played a major role in the severity of CD pseudomembranous colitis and in the occurrence of an inflammatory pseudotumor. PMID- 12581334 TI - Normal and variant arterial supply of the liver and gallbladder. AB - The vascular supply of the liver and gallbladder is characterized by a striking degree of polymorphism. The occurrence of variants that differ from the usual pattern is both surprisingly common and unpredictable. Knowledge of this variable anatomy is particularly important during surgical interventions in which preoperative imaging studies are not available, such as in organ procurements. In this report we consider various anatomical arterial configurations, and describe an unusual variant. PMID- 12581335 TI - Guidelines for the pathoanatomical examination of the lower brain stem in ingestive and swallowing disorders and its application to a dysphagic spinocerebellar ataxia type 3 patient. AB - Despite the fact that considerable progress has been made in the last 20 years regarding the three-phase process of ingestion and the lower brain stem nuclei involved in it, no comprehensive descriptions of the ingestion-related lower brain stem nuclei are available for neuropathologists confronted with ingestive malfunctions. Here, we propose guidelines for the pathoanatomical investigation of these nuclei based on current knowledge with respect to ingestion and the nuclei responsible for this process. The application of these guidelines is described by drawing upon the example of the lower brain stem of a male patient with spinocerebellar ataxia type 3, also known as Machado-Joseph disease, who displayed malfunctions during the preparatory phase of ingestion, as well as lingual and pharyngeal phases of swallowing. By way of the representative application of the recommended investigation procedure to 100 microm serial sections through the patient's brain stem stained for lipofuscin pigment and Nissl material, we observed neuronal loss together with astrogliosis in nearly all of the ingestion-related lower brain stem nuclei (motor, principal and spinal trigeminal nuclei; facial nucleus; parvocellular reticular nucleus; ambiguous nucleus, motor nucleus of the dorsal glossopharyngeal and vagal area; gelatinous, medial, parvocellular and pigmented solitary nuclei; hypoglossal nucleus). In view of their known functional role in the three-phase process of ingestion, damage to these nuclei not only offers an explanation of the patient's malfunctions related to the preparatory phase of ingestion and lingual and pharyngeal phases of swallowing, but also suggests that the patient may have suffered from additional esophageal phase swallowing malfunctions not mentioned in his medical records. PMID- 12581336 TI - Dural haemorrhage in non-traumatic infant deaths: does it explain the bleeding in 'shaken baby syndrome'? AB - A histological review of dura mater taken from a post-mortem series of 50 paediatric cases aged up to 5 months revealed fresh bleeding in the dura in 36/50, the bleeding ranging from small perivascular haemorrhages to extensive haemorrhage which had ruptured onto the surface of the dura. Severe hypoxia had been documented clinically in 27 of the 36 cases (75%). In a similar review of three infants presenting with classical 'shaken baby syndrome', intradural haemorrhage was also found, in addition to subdural bleeding, and we believe that our findings may have relevance to the pathogenesis of some infantile subdural haemorrhage. Recent work has shown that, in a proportion of infants with fatal head injury, there is little traumatic brain damage and that the significant finding is craniocervical injury, which causes respiratory abnormalities, severe global hypoxia and brain swelling, with raised intracranial pressure. We propose that, in such infants, a combination of severe hypoxia, brain swelling and raised central venous pressure causes blood to leak from intracranial veins into the subdural space, and that the cause of the subdural bleeding in some cases of infant head injury is therefore not traumatic rupture of bridging veins, but a phenomenon of immaturity. Hypoxia with brain swelling would also account for retinal haemorrhages, and so provide a unified hypothesis for the clinical and neuropathological findings in cases of infant head injury, without impact or considerable force being necessary. PMID- 12581338 TI - Tau load is associated with apolipoprotein E genotype and the amount of amyloid beta protein, Abeta40, in sporadic and familial Alzheimer's disease. AB - The total amount of hyperphosphorylated tau protein (p-tau load), present as neurofibrillary tangles (NFTs), neuropil threads or plaque neurites, was quantified in the frontal cortex of 109 cases of sporadic Alzheimer's disease (AD) and 35 cases of familial AD due to missense mutations in the presenilin-1, presenilin-2 and amyloid precursor protein genes. p-tau load was inversely correlated with age at onset of illness in both sporadic and familial AD but not with duration of disease. There was no difference in p-tau load between cases of familial AD and others with sporadic AD, matching the familial cases for apolipoprotein E (APO E) genotype. However, p-tau was greater in cases of familial and sporadic AD in the presence of APO E epsilon4 allele and increased with gene dose. Conversely, p-tau load tended to be lower when epsilon2 allele was present. In sporadic AD, tau load was highly significantly correlated with amyloid beta40 (Abeta40), but not Abeta42(43), load. These data indicate that the burden of pathological tau deposited in the brain in both familial and sporadic AD is favoured in the presence of APO E epsilon4 allele and also related to the amount of Abeta40, this also being higher when epsilon4 allele is present. Abeta40 plaques are rich in microglial cells and it is possible that p-tau pathology in AD is triggered by reaction of microglial cells to the presence of Abeta40 and not this peptide directly. PMID- 12581337 TI - Tau phosphorylation and kinase activation in familial tauopathy linked to deln296 mutation. AB - Tau phosphorylation has been examined by immunohistochemistry in the brain of a patient affected with familial tauopathy with progressive supranuclear palsy-like phenotype linked to the delN296 mutation in the tau gene. Phospho-specific tau antibodies Thr181, Ser202, Ser214, Ser396 and Ser422, and antibodies to glycogen synthase kinase-3alpha/beta (GSK-3alpha/beta) and to phosphorylated (P) mitogen activated protein kinase/extracellular signal-regulated kinases (MAPK/ERK), stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), p38 kinase (p38) and GSK-3betaSer9 have been used to gain understanding of the identification of phosphorylation sites, as well as of the specific kinases that regulate tau phosphorylation at those specific sites, in a familial tauopathy. The neuropathological examination disclosed atrophy of the right precentral gyrus and the brainstem. Neurone loss and gliosis were observed in the substantia nigra, several nuclei of the brainstem and diencephalon. Hyper-phosphorylated tau accumulated in neurones with neurofibrillary tangles and in neurones with pretangles in the substantia nigra, locus ceruleus, peri-aqueductal grey matter, reticular formation, motor nuclei of the brainstem, and thalamus, amygdala and hippocampus. tau-immunoreactive astrocytes and, particularly, oligodendrocytes with coiled bodies were widespread in the brainstem, diencephalons, cerebral white matter and cerebral cortex. Increased expression of MAPK/ERK-P, SAPK/JNK-P, p-38-P and GSK-3beta-P was observed in select subpopulations of neurones with neurofibrillary tangles and in neurones with pretangles. MAPK/ERK-P, SAPK/JNK-P, p38-P and GSK-3beta-P were also expressed in tau-containing astrocytes and in oligodendrocytes with coiled bodies. These findings show, for the first time, activation of precise kinases that regulate tau phosphorylation at specific sites in familial tauopathy. PMID- 12581339 TI - Mitochondrial dysfunction in myofibrillar myopathy. AB - 'Myofibrillar myopathy' defines a myopathic condition with focal myofibrillar destruction and accumulation of degraded myofibrillar elements. Despite the fact that a number of mutations in different genes as well as cytotoxic agents lead to the disease, abnormal accumulation of desmin is a typical, common feature. Pathological changes of mitochondrial morphology and function have been observed in animal models with intermediate filament pathology. Therefore, in the present study we tested for mitochondrial pathology in skeletal muscle of five patients with the pathohistological diagnosis of myofibrillar myopathy. Screening for large-scale mtDNA deletions and the frequent MERRF (myoclonic epilepsy; ragged red fibres) and MELAS (mitochondrial encephalomyopathy; lactic acidosis; stroke) point mutations was negative in all patients. Histologically, all muscle biopsies showed nonspecific abnormalities of the oxidative/mitochondrial enzyme stainings (histochemistry for reduced nicotinamide adenine dinucleotide, succinic dehydrogenase, cytochrome c oxidase), only one of them had ragged red fibres and a significant number of cytochrome c oxidase-negative fibres. Upon biochemical investigation, four of our patients showed pathologically low respiratory chain complex I activities. Only one of our patients had a pathologically low complex IV activity, while the measurements of the others were within low normal range. The single patient with pathological values for both complex I and IV was the one with the clear histological hallmarks (ragged red and cytochrome c oxidase negative fibres) of mitochondrial pathology. She also was the only patient with clinical signs hinting at a mitochondrial disorder. Together with data from observations in desmin- and plectin-deficient mice, our results support the view that desmin intermediate filament pathology in these cases is closely linked to mitochondrial dysfunction in skeletal muscle. PMID- 12581340 TI - Transcription factors c-Jun/activator protein-1 and nuclear factor-kappa B in oxidative stress response in mitochondrial diseases. AB - Mitochondrial dysfunction leads to oxygen free radical (ROS) generation with consequent oxidative stress and cellular damage. Recently, activation of the cellular antioxidant system and apoptosis were demonstrated in skeletal muscle fibres from patients with mitochondrial diseases, but the underlying mechanisms remain unknown. Hydrogen peroxide, a by-product of ROS generation, is a chemical inducer of gene expression able to activate apoptosis and to promote the antioxidant response through the activation of nuclear factor-kappa B (NF-kappaB) and activator protein-1 (AP-1) transcription factor. Using immunohistochemistry and confocal microscopy, we evaluated the expression of NF-kappaB and AP-1 in muscle biopsies from patients with mitochondrial disease. In addition, we examined the expression of factors involved in their activation, such as NF kappaB inducing kinase (NIK) and phosphorylated Jun-N-terminal kinase (p-JNK). Most fibres with respiratory chain dysfunction displayed nuclear staining for activated c-Jun/AP-1, but not for NF-kappaB. The same fibres reacted for p-JNK. Only some ragged red fibres immunoreacted for NIK. These data suggest that AP-1 is involved in the oxidative stress response in muscle fibres from patients with mitochondrial disease. PMID- 12581341 TI - Involvement of precerebellar nuclei in multiple system atrophy. AB - In this semiquantitative study based on 26 post-mortem cases, we describe the involvement of precerebellar nuclei in multiple system atrophy (MSA), a progressive degenerative disorder of the human central nervous system characterized by abnormal, argyrophilic and alpha-synuclein immunopositive intracellular inclusions within selectively vulnerable oligodendrocytes and nerve cells. The Campbell-Switzer silver-pyridine technique with alpha-synuclein immunoreactions using 100-microm thick sections is recommended over more conventional methods, thereby permitting visualization of the pertinent lesions in greater detail and facilitating post-mortem diagnosis of MSA specimens. Affected oligodendrocytes occur in specific fibre tracts and grey matters, with most pathology being observed in projections from the precerebellar nuclei to the cerebellum (ponto-cerebellar, olivo-cerebellar, reticulo-cerebellar tracts) and in descending/ascending fibre tracts of the motor system (cortico-pontine, cortico-bulbar, cortico-spinal, spino-reticular, spino-olivary, spino-cerebellar tracts). Three types of abnormal intraneuronal aggregations occur: (i) a loosely woven network within the cell nucleus; (ii) a latticework accumulating in peripheral portions of the cell body; and (iii) irregularly outlined patches of compact, intensely argyrophilic material usually located within deposits of lipofuscin granules. Counter-staining for the presence of extraneuronal lipofuscin can aid neuropathologists in the recognition of lost existent neurones in MSA. Neurones with inclusion bodies occur in the inferior olivary nuclear complex, lateral reticular nucleus, external cuneate nucleus, conterminal nucleus, interfascicular nucleus, nucleus of Roller, dorsal paramedian reticular nucleus, subventricular nucleus, arcuate nucleus, pontobulbar body and pontine grey. The lateral reticular nucleus and accessory nuclei of the inferior olive sustain the most damage and reveal prominent neuronal loss, followed by the pontobulbar body and arcuate nucleus. The uniformly bilateral damage and, in some cases, even obliteration of the nuclei studied, supply additional evidence for the pathoanatomical substrata of the cerebellar dysfunction that reportedly emerges in the clinical course of MSA. PMID- 12581342 TI - An atypical form of sporadic panencephalopathic Creutzfeldt-Jakob disease in Japan. PMID- 12581343 TI - Apolipoprotein E genotype in senile dementia with tangles differs from Alzheimer's disease. PMID- 12581346 TI - Microarray expression profiling: capturing a genome-wide portrait of the transcriptome. AB - The bacterial transcriptome is a dynamic entity that reflects the organism's immediate, ongoing and genome-wide response to its environment. Microarray expression profiling provides a comprehensive portrait of the transcriptional world enabling us to view the organism as a 'system' that is more than the sum of its parts. The vigilance of microorganisms to environmental change, the alacrity of the transcriptional response, the short half-life of bacterial mRNA and the genome-scale nature of the investigation collectively explain the power of this method. These same features pose the most significant experimental design and execution issues which, unless surmounted, predictably generate a distorted image of the transcriptome. Conversely, the expression profile of a properly conceived and conducted microarray experiment can be used for hypothesis testing: disclosure of the metabolic and biosynthetic pathways that underlie adaptation of the organism to chang-ing conditions of growth; the identification of co ordinately regulated genes; the regulatory circuits and signal transduction systems that mediate the adaptive response; and temporal features of developmental programmes. The study of bacterial pathogenesis by microarray expression profiling poses special challenges and opportunities. Although the technical hurdles are many, obtaining expression profiles of an organism growing in tissue will probably reveal strategies for growth and survival in the host's microenvironment. Identifying these colonization strategies and their cognate expression patterns involves a 'deconstruction' process that combines bioinformatics analysis and in vitro DNA array experimentation. PMID- 12581345 TI - Differential analysis of DNA microarray gene expression data. AB - Here, we review briefly the sources of experimental and biological variance that affect the interpretation of high-dimensional DNA microarray experiments. We discuss methods using a regularized t-test based on a Bayesian statistical framework that allow the identification of differentially regulated genes with a higher level of confidence than a simple t-test when only a few experimental replicates are available. We also describe a computational method for calculating the global false-positive and false-negative levels inherent in a DNA microarray data set. This method provides a probability of differential expression for each gene based on experiment-wide false-positive and -negative levels driven by experimental error and biological variance. PMID- 12581347 TI - VirD4-independent transformation by CloDF13 evidences an unknown factor required for the genetic colonization of plants via Agrobacterium. AB - Agrobacterium uses a mechanism similar to conjugation for trans-kingdom transfer of its oncogenic T-DNA. A defined VirB/VirD4 Type IV secretion system is responsible for such a genetic transfer. In addition, certain virulence proteins as VirE2 can be mobilized into host cells by the same apparatus. VirE2 is essential to achieve plant but not yeast transformation. We found that the limited host range plasmid CloDF13 can be recruited by the virulence apparatus of Agrobacterium for transfer to eukaryotic hosts. As expected the VirB transport complex was required for such trans-kingdom DNA transfer. However, unexpectedly, the coupling factor VirD4 turned out to be necessary for transfer to plants but not for transport into yeast. The CloDF13 encoded coupling factor (Mob) was essential for transfer to both plants and yeast though. This is interpreted by the different specificities of Mob and VirD4. Hence, Mob being required for the transport of the CloDF13 transferred DNA (to both plants and yeast) and VirD4 being required for transport of virulence proteins such as VirE2. Nevertheless, the presence of the VirE2 protein in the host plant was not sufficient to restore the deficiency for VirD4 in the transforming bacteria. We propose that Mob functions encoded by the plasmid CloDF13 are sufficient for DNA mobilization to eukaryotic cells but that the VirD4-mediated pathway is essential to achieve DNA nuclear establishment specifically in plants. This suggests that other Agrobacterium virulence proteins besides VirE2 are translocated and essential for plant transformation. PMID- 12581348 TI - Architecture of a protein central to iron homeostasis: crystal structure and spectroscopic analysis of the ferric uptake regulator. AB - Iron is an essential element for almost all organisms, although an overload of this element results in toxicity because of the formation of hydroxyl radicals. Consequently, most living entities have developed sophisticated mechanisms to control their intracellular iron concentration. In many bacteria, including the opportunistic pathogen Pseudomonas aeruginosa, this task is performed by the ferric uptake regulator (Fur). Fur controls a wide variety of basic physiological processes including iron uptake systems and the expression of exotoxin A. Here, we present the first crystal structure of Fur from P. aeruginosa in complex with Zn2+ determined at a resolution of 1.8 A. Furthermore, X-ray absorption spectroscopic measurements and microPIXE analysis were performed in order to characterize the distinct zinc and iron binding sites in solution. The combination of these complementary techniques enables us to present a model for the activation and DNA binding of the Fur protein. PMID- 12581349 TI - A plasmid-encoded regulator couples the synthesis of toxins and surface structures in Bacillus anthracis. AB - Transcription of the major Bacillus anthracis virulence genes is triggered by CO2, a signal believed to reflect the host environment. A 180 kb plasmid, pXO1, carries the anthrax toxin genes and the genes responsible for their regulation, pagR and atxA; the latter encodes a major trans-activator. It has long been known that pXO1 genes have major effects on the physiology of B. anthracis, probably through regulatory cross-talk between plasmid and chromosomal genes. Accordingly, we found that the chromosomal S-layer genes, sap and eag, are regulated by pXO1 genes so that only eag is significantly expressed in the presence of CO2. This effect results from the product of pagR acting as the most downstream element of a signalling cascade initiated by AtxA. In vitro evidence showed that PagR is a transcription factor that controls the S-layer genes by direct binding on their promoter regions. This work provides evidence that AtxA is a master regulator that co-ordinates the response to host signals by orchestrating positive and negative controls over genes located on all genetic elements. PMID- 12581350 TI - The Caulobacter crescentus polar organelle development protein PodJ is differentially localized and is required for polar targeting of the PleC development regulator. AB - Regulation of polar development and cell division in Caulobacter crescentus relies on the dynamic localization of several proteins to cell poles at specific stages of the cell cycle. The polar organelle development protein, PodJ, is required for the synthesis of the adhesive holdfast and pili. Here we show the cell cycle localization of PodJ and describe a novel role for this protein in controlling the dynamic localization of the developmental regulator PleC. In swarmer cells, a short form of PodJ is localized at the flagellated pole. Upon differentiation of the swarmer cell into a stalked cell, full length PodJ is synthesized and localizes to the pole opposite the stalk. In late predivisional cells, full length PodJ is processed into a short form which remains localized at the flagellar pole after cell division and is degraded during swarmer to stalked cell differentiation. Polar localization of the developmental regulator PleC requires the presence of PodJ. In contrast, the polar localization of PodJ is not dependent on the presence of PleC. These results indicate that PodJ is an important determinant for the localization of a major regulator of cell differentiation. Thus, PodJ acts directly or indirectly to target PleC to the incipient swarmer pole, to establish the cellular asymmetry that leads to the synthesis of holdfasts and pili at their proper subcellular location. PMID- 12581351 TI - A screen in Saccharomyces cerevisiae identified CaMCM1, an essential gene in Candida albicans crucial for morphogenesis. AB - Morphogenesis in Saccharomyces cerevisiae and the pathogenic yeast Candida albicans is governed in part by the same molecular circuits. In S. cerevisiae, FLO11/MUC1 expression has been shown to be modulated by multiple signalling pathways required for pseudohyphal development. We have established a screen in S. cerevisiae to identify regulators of fungal development in C. albicans based on FLO11::lacZ expression as a reporter. This screen identified both known components of the mitogen-activated protein kinase (MAPK) cascade and the cAMP cascade that are important for hyphal development in C. albicans, as well as genes not yet known to be involved in morphogenesis. The Candida homologue of MCM1 is one of the novel factors identified in this screen as being important for morphogenesis. CaMcm1p levels do not vary significantly in different cell types and respond to an autoregulatory feedback mechanism, arguing that CaMcm1p activity is regulated by post-translational modifications. Both overexpression and repression of this essential gene led to the induction of hyphae. Moreover, we found that the expression of HWP1, a hyphae-specific gene, was induced by repression of CaMCM1. The changes in morphology and HWP1 expression were not the result of a change in expression levels of NRG1 or TUP1, known repressors of hyphal development. Thus, CaMcm1p is a component of a hitherto unknown regulatory mechanism of hyphal growth. PMID- 12581352 TI - The modular structure of Escherichia coli threonyl-tRNA synthetase as both an enzyme and a regulator of gene expression. AB - In addition to its role in tRNA aminoacylation, Escherichia coli threonyl-tRNA synthetase is a regulatory protein which binds a site, called the operator, located in the leader of its own mRNA and inhibits translational initiation by competing with ribosome binding. This work shows that the two essential steps of regulation, operator recognition and inhibition of ribosome binding, are performed by different domains of the protein. The catalytic and the C-terminal domain of the protein are involved in binding the two anticodon arm-like structures in the operator whereas the N-terminal domain of the enzyme is responsible for the competition with the ribosome. This is the first demonstration of a modular structure for a translational repressor and is reminiscent of that of transcriptional regulators. The mimicry between the operator and tRNA, suspected on the basis of previous experiments, is further supported by the fact that identical regions of the synthetase recognize both the operator and the tRNA anticodon arm. Based on these results, and recent structural data, we have constructed a computer-derived molecular model for the operator-threonyl-tRNA synthetase complex, which sheds light on several essential aspects of the regulatory mechanism. PMID- 12581353 TI - AREA directly mediates nitrogen regulation of gibberellin biosynthesis in Gibberella fujikuroi, but its activity is not affected by NMR. AB - AREA (NIT2) is a general transcription factor involved in derepression of numerous genes responsible for nitrogen utilization in Gibberella fujikuroi and many other fungi. We have previously shown that the deletion of areA-GF resulted in mutants with significantly reduced gibberellin (GA) production. Here we demonstrate that the expression level of six of the seven GA biosynthesis genes is drastically reduced in mutants lacking areA. Furthermore, we show that, despite the fact that GAs are nitrogen-free diterpenoid compounds, which are not obviously involved in nitrogen metabolism, AREA binds directly to the promoters of the six N-regulated genes. The binding of AREA was analysed in more detail using the promoter of one of the GA-biosynthesis genes encoding the ent-kaurene oxidase (P450-4). Deletion/mutation analysis of the P450-4 promoter fused to the Escherichia coli uidA gene, which encodes beta-glucuronidase, allowed the in vivo identification of functional GATA motifs. We have also analysed the nmr gene of G. fujikuroi (nmr-GF) which has high similarity to the Neurospora crassa nmr-1 and Aspergillus nidulans nmrA genes, both involved in nitrogen metabolite repression. In contrast to our expectation, deletion of nmr-GF did not result in significant derepression of the GA biosynthesis genes in the presence of ammonium, glutamine or glutamate. Overexpression of the nmr-GF gene fused to the strong promoter of the G. fujikuroi glutamine synthetase (gs) gene revealed only a very slight repression of the nitrate reductase (niaD) gene, resulting in weak resistance to chlorate. Surprisingly, this effect was only observed in the presence of high amounts of glutamate; cultivation on ammonium failed to induce any resistance to chlorate. Despite the limited effect of gene replacement and overexpression of nmr-GF on the nitrogen metabolism of G. fujikuroi itself, the gene fully restored nitrogen metabolite repression in A. nidulans and N. crassa nmr mutants. Therefore, we postulate that, in contrast to A. nidulans and N. crassa, NMR does not function independently as the main modulator of AREA in G. fujikuroi. PMID- 12581354 TI - Identification of a novel Gsp-related pathway required for secretion of the manganese-oxidizing factor of Pseudomonas putida strain GB-1. AB - The manganese-oxidizing factor of Pseudomonas putida strain GB-1 is associated with the outer membrane. One of the systems of protein transport across the outer membrane is the general secretory pathway (Gsp). The gsp genes are called xcp in Pseudomonas species. In a previous study, it was shown that mutation of the prepilin peptidase XcpA and of a homologue of the pseudopilin XcpT inhibited transport of the factor. In the present study, we describe the genomic region flanking the xcpT homologue (designated xcmT1). We show that xcmT1 is part of a two-gene operon that includes an xcpS homologue (designated xcmS). No other xcp like genes are present in the regions flanking the xcmT1/xcmS cluster. We also characterized the site of transposon insertion of another transport mutant of P. putida GB-1. This insertion appeared to be located in a gene (designated xcmX) possibly encoding another pseudopilin-related protein. This xcmX is clustered with two other xcpT-related genes (designated xcmT2 and xcmT3) on one side and homologues of three csg genes (designated csmE, csmF and csmG) on the other side. The csg genes are involved in production of aggregative fibres in Escherichia coli and Salmonella typhimurium. A search for XcmX homologues revealed that the recently published genome of Ralstonia solanacearum and the unannotated genome of P. putida KT2440 contain comparable gene clusters with xcmX and xcp homologues that are different from the well-described 'regular'xcp/gsp clusters. They do contain xcpR and xcpQ homologues but, for example, homologues of xcpP, Y and Z are lacking. The results suggest a novel Xcp-related system for the transport of manganese-oxidizing enzymes to the cell surface. PMID- 12581355 TI - Copper-mediated reversal of defective laccase in a Deltavph1 avirulent mutant of Cryptococcus neoformans. AB - Previous studies have shown that a Deltavph1 Cryptococcus neoformans mutant defective in vesicular acidification lacked several important virulence factors including a copper-containing laccase and was avirulent in a mouse model. In the present studies, we characterized laccase transcription and protein production to obtain insights into the mechanism of the vph1 mutation in this pathogen. Although transcription and protein expression were somewhat reduced, laccase protein was found to be successfully translated and correctly targeted to the cell wall in the Deltavph1 mutant as shown by Western blot and immuno-electron microscopy, despite a complete lack of laccase activity. Laccase activity was substantially restored in metabolically active Deltavph1 cells at 30 degrees C by addition of 100 micro M copper sulphate. This restoration by copper was found to occur through both transcriptional and post-translational mechanisms. Laccase transcriptional induction by copper was found to be dependent on enhancer region II within the 5'-untranslated region of CNLAC1. Copper was also found to restore partial activity to Deltavph1 cells at 0 degrees C, suggesting that cell wall laccase was expressed in the mutant as an apo-enzyme. Apo-laccase restoration by copper was found to be facilitated by an acidic environment, consistent with a role for the vacuolar (H+)-ATPase proton pump in copper assembly of laccase in C. neoformans. PMID- 12581357 TI - Ssn6, an important factor of morphological conversion and virulence in Candida albicans. AB - Candida albicans, the major fungal pathogen in humans, undergoes morphological conversion from yeasts to filamentous growth forms depending upon various environmental conditions. Here, we have identified a C. albicans gene, namely SSN6, encoding a putative global transcriptional co-repressor that is highly homologous to the Saccharomyces cerevisiae Ssn6. The isolated C. albicans SSN6 complemented the pleiotropic phenotypes of S. cerevisiae ssn6 mutation, and its expression levels declined significantly in response to a strong true hyphal inducer, serum. The mutant lacking C. albicans Ssn6 displayed a stubby pseudohyphal growth pattern, derepressed filament-specific genes in response to elevated temperature 37 degrees C and failed to develop true hyphae, extensive filamentation and virulence. Such morphological defects of ssn6/ssn6 mutant were not rescued by overexpression of Tup1, Cph1 or Efg1. Moreover, epistatic analysis showed that, as far as cell morphology was concerned, Ssn6 was epistatic to Tup1 at the higher temperature but that, at the lower temperature, the ssn6/ssn6 tup1/tup1 double mutant grew in a stubby form of pseudohyphae distinct from the phenotypes of either single mutant. Furthermore, overexpression of SSN6 in C. albicans led to enhanced filamentous growth and attenuated virulence. These findings suggest that Ssn6 may function as an activator as well as a repressor of filamentous growth and be a target for candidacidal drugs, as its excess or deficiency resulted in impaired virulence. PMID- 12581356 TI - Versatility of inner membrane protein biogenesis in Escherichia coli. AB - To further our understanding of inner membrane protein (IMP) biogenesis in Escherichia coli, we have accomplished the widest in vivo IMP assembly screen so far. The biogenesis of a set of model IMPs covering most IMP structures possible has been studied in a variety of signal recognition particle (SRP), Sec and YidC mutant strains. We show that the assembly of the complete set of model IMPs is assisted (i.e. requires the aid of proteinaceous factors), and that the requirements for assembly of the model IMPs into the inner membrane differ significantly from each other. This indicates that IMP assembly is much more versatile than previously thought. PMID- 12581358 TI - Transcriptional organization and regulation of the Escherichia coli K30 group 1 capsule biosynthesis (cps) gene cluster. AB - Escherichia coli group 1 capsules are important virulence determinants, yet little is known about the transcriptional organization or regulation of their biosynthetic (cps) operons. Transcription of the prototype serotype K30 cluster is modulated by the JUMPStart-RfaH antitermination mechanism, with the cps promoter being localized to a region immediately upstream of the JUMPStart sequence. A putative stem-loop structure located within the K30 cps cluster separates conserved genes with products that are required for surface expression of capsule from serotype-specific genes encoding enzymes for polymer repeat-unit synthesis and polymerization. This putative stem-loop structure significantly reduces transcription in a termination-probe vector and may contribute to differential expression of the cps genes. Previous work indicated that increased amounts of group 1 capsular polysaccharide synthesis resulted from the overexpression of the Rcs (regulator of capsule synthesis) proteins. However, neither overexpression of the transcriptional activator RcsB nor an rcsB::aadA chromosomal insertion altered the level of transcription measured by cps::lacZ fusions. In the group 1 strains examined, an RcsAB box was found immediately upstream of galF, a gene involved in the production of sugar nucleotide precursors. Overexpression of RcsB was found to result in a threefold increase in transcription of a galF::lacZ chromosomal fusion. Moreover, overexpression of GalF gave rise to a two- to threefold increase in cell-free as well as cell associated capsule, without affecting cps::lacZ activity. These results indicate that transcription of the E. coli group 1 capsule cluster itself is not regulated by the Rcs system and may, in fact, be constitutive. However, the Rcs system can potentially influence levels of capsular polysaccharide production by increasing galF transcription and influencing the available pool of biosynthetic precursors. PMID- 12581359 TI - Comparative genomics reveal novel heat shock regulatory mechanisms in Staphylococcus aureus and other Gram-positive bacteria. AB - Multiple regulatory mechanisms for coping with stress co-exist in low G+C Gram positive bacteria. Among these, the HrcA and CtsR repressors control distinct regulons in the model organism, Bacillus subtilis. We recently identified an orthologue of the CtsR regulator of stress response in the major pathogen, Staphylococcus aureus. Sequence analysis of the S. aureus genome revealed the presence of potential CtsR operator sites not only upstream from genes encoding subunits of the Clp ATP-dependent protease, as in B. subtilis, but also, unexpectedly, within the promoter regions of the dnaK and groESL operons known to be specifically controlled by HrcA. The tandem arrangement of the CtsR and HrcA operators suggests a novel mode of dual heat shock regulation by these two repressors. The S. aureus ctsR and hrcA genes were cloned under the control of the PxylA xylose-inducible promoter and used to demonstrate dual regulation of the dnaK and groESL operons by both CtsR and HrcA, using B. subtilis as a heterologous host. Direct binding by both repressors was shown in vitro by gel mobility shift and DNase I footprinting experiments using purified S. aureus CtsR and HrcA proteins. DeltactsR, DeltahrcA and DeltactsRDeltahrcA mutants of S. aureus were constructed, indicating that the two repressors are not redundant but, instead, act together synergistically to maintain low basal levels of expression of the dnaK and groESL operons in the absence of stress. This novel regulatory mode appears to be specific to Staphylococci. PMID- 12581360 TI - Interaction of the sensor module of Mycobacterium tuberculosis H37Rv KdpD with members of the Lpr family. AB - The genetic and biochemical mechanisms by which Mycobacterium tuberculosis senses and responds to the complex environment that it encounters during infection and persistence within the host remain unknown. In a number of bacterial species, the Kdp signal transduction pathway appears to be the primary response to environmental osmotic stress, which is primarily mediated by K+ concentration in bacteria. We show that kdp encodes for components of a mycobacterial signalling pathway by demonstrating the K+ dependence of kdpFABC expression in both M. tuberculosis H37Rv and Mycobacterium smegmatis. To identify proteins of M. tuberculosis that participate in this signalling pathway, we used the N-terminal sensing module of the histidine kinase KdpD as bait in a yeast two-hybrid screen. We show that the sensing domain of KdpD interacts specifically with two membrane lipoproteins, LprJ (Rv1690) and LprF (Rv1368). Overexpression of lprF and lprJ alleles in mycobacterial kdpF-lacZ reporter strains enabled us to identify alleles that modulate kdpFABC expression. By exploiting the yeast three-hybrid system, we have found that the histidine kinase domain of KdpD forms ternary complexes with LprF and LprJ and the sensing module of KdpD. Our results establish a role for membrane proteins in the Kdp signalling pathway and suggest that LprF and LprJ function as accessory or ligand-binding proteins that communicate directly with the sensing domain of KdpD to modulate kdp expression. PMID- 12581361 TI - PriA supports two distinct pathways for replication restart in UV-irradiated Escherichia coli cells. AB - The Escherichia coli PriA protein loads the DnaB replicative helicase at branched DNA structures independently of the replication initiator protein, DnaA, and thereby facilitates assembly of functional replisomes at sites removed from oriC. It is therefore a critical factor in the rescue of replication forks stalled at DNA lesions. It is also a DNA helicase. We describe insertions near the 3' end of priA that interfere with PriA activity. These insertions and the previously described priA300 encoding helicase-defective PriA K230R are shown to be effective suppressors of the DNA repair defect in recG strains, but substantially reduce the ability of ruv mutants to survive DNA damage. The data presented suggest that PriA helicase in conjunction with RecG can promote direct rescue of stalled forks independently of the recombinational pathway promoted by the combined activities of the RuvABC, RecBCD and RecA proteins, which requires only the primosome assembly activity of PriA to load DnaB at D loops. In cells lacking the helicase activity of PriA, we propose that stalled forks can be redirected to the recombination pathway via a Holliday junction intermediate common to both pathways, thus explaining the resistance of these cells to DNA damage. PMID- 12581362 TI - Identification and characterization of genomic loci unique to the Brazilian purpuric fever clonal group of H. influenzae biogroup aegyptius: functionality explored using meningococcal homology. AB - Brazilian purpuric fever (BPF) is a fulminant septicaemic infection of young children, caused by a clonal group of strains of Haemophilus influenzae biogroup aegyptius (Hae), an organism previously solely associated with conjunctivitis. Their special capacity to invade from the initial site of conjunctival infection is unexplained. A polymerase chain reaction (PCR)-amplified subtractive hybridization technique was used to identify genes specific to the BPF clonal group. A copy of bacteriophage HP1 and 46 further chromosomal loci were identified in the BPF but not in the conjunctivitis strain of Hae. Sixteen were characterized further, and one - encoding an analogue of the Legionella pneumophila epithelial cell entry-enhancing protein EnhC - was investigated in depth. Two genes, bpf001 and bpf002, unique to the BPF clonal group were identified between homologues of HI1276 and HI1277 in a complex locus close to H. influenzae genetic island 1, recently identified in pathogenic H. influenzae type b. Bpf001 encodes a protein homologous to EnhC and to the previously uncharacterized product of the meningococcal gene NMB0419. Functional studies of bpf001 proving intractable, NMB0419 was chosen as a surrogate for investigation and shown to modulate bacterial interaction with monolayers of human respiratory epithelial cells, promoting invasion, the first stage (for Hae) in the pathogenesis of BPF. PMID- 12581363 TI - Identification of a protein, YneA, responsible for cell division suppression during the SOS response in Bacillus subtilis. AB - A knock-out mutant of the dinR gene that encodes the SOS regulon repressor in Bacillus subtilis was constructed. The yneA, yneB and ynzC genes transcribed divergently from the dinR gene were strongly induced in mutant cells. Northern hybridization analyses revealed that these genes collectively form an operon and belong to the SOS regulon. The simultaneous deletion of dinR and yneA suppressed the filamentous phenotype of the dinR mutant. Furthermore, although yneA is suppressed in the wild-type cell in the absence of SOS induction, artificial expression of the YneA protein using an IPTG-inducible promoter resulted in cell elongation. Disruption of yneA significantly reduced cell elongation after the induction of the SOS response by mitomycin C in dinR+ cells. These results indicate that the YneA protein is responsible for cell division suppression during the SOS response in B. subtilis. Localization of the FtsZ protein to the cell division site was reduced in dinR-disrupted or yneA-expressing cells, further suggesting that the YneA protein suppresses cell division through the suppression of FtsZ ring formation. Interestingly, the B. subtilis YneA protein is structurally and phylogenetically unrelated to its functional counterpart in Escherichia coli, SulA. PMID- 12581364 TI - The dual roles of AlgG in C-5-epimerization and secretion of alginate polymers in Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa strains causing chronic pulmonary infections in cystic fibrosis patients produce high levels of alginate, an exopolysaccharide that confers a mucoid phenotype. Alginate is a linear polymer of d-mannuronate (M) and variable amounts of its C-5-epimer, l-guluronate (G). AlgG is a periplasmic C-5 epimerase that converts poly d-mannuronate to the mixed M+G sequence of alginate. To understand better the role and mechanism of AlgG activity, a mutant was constructed in the mucoid strain FRD1 with a defined non-polar deletion of algG. Instead of producing poly mannuronate, the algG deletion mutant secreted dialysable uronic acids, as does a mutant lacking the periplasmic protein AlgK. High levels of unsaturated ends and the nuclear magnetic resonance spectroscopy pattern revealed that the small, secreted uronic acids were the products of extensive polymer digestion by AlgL, a periplasmic alginate lyase co-expressed with AlgG and AlgK. Thus, AlgG is bifunctional with (i) epimerase activity and (ii) a role in protecting alginate from degradation by AlgL during transport through the periplasm. AlgK appears to share the second role. AlgG and AlgK may be part of a periplasmic protein complex, or scaffold, that guides alginate polymers to the outer membrane secretin (AlgE). To characterize the epimerase activity of AlgG further, the algG4 allele of poly mannuronate-producing FRD462 was shown to encode a protein lacking only the epimerase function. The sequence of algG4 has a Ser-272 to Asn substitution in a serine-threonine-rich and conserved region of AlgG, which revealed a critical residue for C-5-epimerase activity. PMID- 12581365 TI - Osmoregulation in Lactococcus lactis: BusR, a transcriptional repressor of the glycine betaine uptake system BusA. AB - The busA (opuA) locus of Lactococcus lactis encodes a glycine betaine uptake system. Transcription of busA is osmotically inducible and its induction after an osmotic stress is reduced in the presence of glycine betaine. Using a genetic screen in CLG802, an Escherichia coli strain carrying a lacZ transcriptional fusion expressed under the control of the busA promoter, we isolated a genomic fragment from the L. lactis subsp. cremoris strain MG1363, which represses transcription from busAp. The cloned locus responsible for this repression was identified as a gene present upstream from the busA operon, encoding a putative DNA binding protein. This gene was named busR. Electrophoretic mobility shift and footprinting experiments showed that BusR is able to bind a site that overlaps the busA promoter. Overexpression of busR in L. lactis reduced expression of busA. Its disruption led to increased and essentially constitutive transcription of busA at low osmolarity. Therefore, BusR is a major actor of the osmotic regulation of busA in L. lactis. PMID- 12581366 TI - Activation mechanisms of the HAC1-mediated unfolded protein response in filamentous fungi. AB - The unfolded protein response (UPR) is a regulatory pathway activating genes involved in multiple functions related to folding, quality control and transport of secreted proteins. Characterization of the hac1/hacA genes encoding the UPR transcription factors from the filamentous fungi Trichoderma reesei and Aspergillus nidulans is described in this article. The corresponding gene in Saccharomyces cerevisiae is activated through a non-spliceosomal intron-splicing reaction. The T. reesei hac1 and A. nidulans hacA mRNAs undergo an analogous splicing reaction of a 20-nt-long intron during UPR induction. This splicing changes the reading frame of the mRNA and thus could bring in an activation domain to the HACI/HACA proteins. In addition to the non-spliceosomal splicing, the hac1/A mRNAs of the filamentous fungi are truncated at the 5'-flanking region upon UPR induction. An upstream open reading frame is omitted from the mRNAs due to the truncation, and evidence is presented showing that the truncated T. reesei hac1 mRNA is translated more efficiently than a full-length mRNA. This paper reports a novel combination of two different regulatory mechanisms of a transcription factor gene, both operational at the mRNA level. PMID- 12581369 TI - The novel method for isolating chitinolytic bacteria and its application in screening for hyperchitinase producing mutant of Alcaligenes xylosoxydans. AB - AIMS: To develop a novel, rapid and effective screening method for chitinase producing bacteria. METHODS AND RESULTS: A simple and rapid technique for screening of potential chitinolytic bacteria has been developed using the chitin binding dye calcofluor white M2R in chitin agar. Microorganisms possessing high chitinolytic potential gave a clear zone under ultraviolet light after 24-48 h of incubation. This method was successfully applied for isolating the hyperchitinase mutant of Alcaligenes xylosoxydans. The mutant Alc. xylosoxydans EMS 33 was found to produce 3.4 times more chitinase than the wild type. CONCLUSIONS: In this study, the screening method for chitinase producing bacteria has been developed and it was applied to screen chitinase-overproducing mutant of Alc. xylosoxydans. SIGNIFICANCE AND IMPACT OF THE STUDY: The novel screening method for chitinase producer is more sensitive, rapid, user-friendly and reliable, which can also be used for screening of recombinants having chitinase gene. PMID- 12581367 TI - Inorganic phosphate is sensed by specific phosphate carriers and acts in concert with glucose as a nutrient signal for activation of the protein kinase A pathway in the yeast Saccharomyces cerevisiae. AB - Yeast cells starved for inorganic phosphate on a glucose-containing medium arrest growth and enter the resting phase G0. We show that re-addition of phosphate rapidly affects well known protein kinase A targets: trehalase activation, trehalose mobilization, loss of heat resistance, repression of STRE-controlled genes and induction of ribosomal protein genes. Phosphate-induced activation of trehalase is independent of protein synthesis and of an increase in ATP. It is dependent on the presence of glucose, which can be detected independently by the G-protein coupled receptor Gpr1 and by the glucose-phosphorylation dependent system. Addition of phosphate does not trigger a cAMP signal. Despite this, lowering of protein kinase A activity by mutations in the TPK genes strongly reduces trehalase activation. Inactivation of phosphate transport by deletion of PHO84 abolishes phosphate signalling at standard concentrations, arguing against the existence of a transport-independent receptor. The non-metabolizable phosphate analogue arsenate also triggered signalling. Constitutive expression of the Pho84, Pho87, Pho89, Pho90 and Pho91 phosphate carriers indicated pronounced differences in their transport and signalling capacities in phosphate-starved cells. Pho90 and Pho91 sustained highest phosphate transport but did not sustain trehalase activation. Pho84 sustained both transport and rapid signalling, whereas Pho87 was poor in transport but positive for signalling. Pho89 displayed very low phosphate transport and was negative for signalling. Although the results confirmed that rapid signalling is independent of growth recovery, long term mobilization of trehalose was much better correlated with growth recovery than with trehalase activation. These results demonstrate that phosphate acts as a nutrient signal for activation of the protein kinase A pathway in yeast in a glucose-dependent way and they indicate that the Pho84 and Pho87 carriers act as specific phosphate sensors for rapid phosphate signalling. PMID- 12581370 TI - Induction of intestinal IgA and IgG antibodies preventing adhesion of verotoxin producing Escherichia coli to Caco-2 cells by oral immunization with liposomes. AB - AIMS: To examine the efficacy of liposome oral administration to induce systemic and mucosal immune responses against verotoxin-producing Escherichia coli (VTEC) and the effect of the induced antibodies on the binding of the bacteria to Caco-2 cells. METHODS AND RESULTS: Mice were immunized orally with VTEC antigen and monophosphoryl lipid A (MPL)-containing liposomes composed of dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylserine and cholesterol (1 : 1 : 2, molar ratio) (PS-liposome). After immunization, significant IgA and IgG responses to VTEC were induced in both serum and the intestinal lavage fluid in all mice tested. Furthermore, anti-VTEC IgA and IgG antibodies in the lavage fluid effectively inhibited the adhesion of VTEC to Caco-2 cells. CONCLUSIONS: Oral immunization with liposome-associated E. coli O157:H7 antigen can induce significant systemic and mucosal antibody responses against the bacterial antigen and antibodies produced in the intestinal tract, thus functioning as inhibitors for preventing VTEC infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Oral PS liposome vaccines containing MPL have the potential usefulness for the induction of a protective mucosal immune response against intestinal diseases. PMID- 12581371 TI - Pulsed electric fields cause sublethal injury in Escherichia coli. AB - AIMS: The objective was to investigate the occurrence of sublethal injury in Escherichia coli by pulsed electric fields (PEF) at different pH values. METHODS AND RESULTS: The occurrence of sublethal injury in PEF-treated E. coli cells depended on the pH of the treatment medium. Whereas a slight sublethal injury was detected at pH 7, 99.95% of survivors were injured when cells were treated at pH 4 for 400 micros at 19 kV. The PEF-injured cells were progressively inactivated by a subsequent holding at pH 4. CONCLUSIONS: PEF cause sublethal injury in E. coli. The measurement of sublethal injury using a selective medium plating technique allowed prediction of the number of cells that would be inactivated by subsequent storage in acidic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This work could be useful for improving food preservation by PEF technology and contributes to the knowledge of the mechanism of microbial inactivation by PEF. PMID- 12581372 TI - Digoxigenin-labelled peptide nucleic acid to detect lactobacilli PCR amplicons immobilized on membranes from denaturing gradient gel electrophoresis. AB - AIMS: To develop a digoxigenin (DIG)-labeled peptide nucleic acid (PNA) probe for the detection of Lactobacillus-related genera amongst eubacterial amplicons obtained from vaginal samples using denaturing gradient gel electrophoresis (DGGE) blots. METHODS AND RESULTS: Part of the 16S rRNA gene sequence was used as a target for the PNA probe. After confirming probe specificity using chromosomal DNA from species and isolates that have been detected in the urogenital tract, it was successfully used to detect lactobacilli amplicons generated using eubacterial-specific 16S rRNA gene-targeted primers from vaginal tract samples immobilized on membranes from DGGE. CONCLUSIONS: The Lactobacillus-specific PNA probe could distinguish between DNA fragments from lactobacilli in a DGGE gel from other bacterial species, including those that migrated to a similar position. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of the DIG-labelled PNA probe on blots of eubacterial PCR products from DGGE gels can be used to specifically detect lactobacilli in complex vaginal samples. PMID- 12581373 TI - A comparison of thiosulphate-citrate-bile salts-sucrose (TCBS) agar and thiosulphate-chloride-iodide (TCI) agar for the isolation of Vibrio species from estuarine environments. AB - AIMS: The purpose of this study was to compare a recently described medium, thiosulphate-chloride-iodide (TCI), for the isolation of estuarine vibrios with thiosulphate-citrate-bile salts-sucrose (TCBS). METHODS: A total of 492 colonies which developed on these media from estuarine water samples taken monthly over a 10-month period were examined. RESULTS: A much larger number of colonies developed on TCBS than TCI, and minimal taxonomic criteria indicated that a higher percentage (61%) of TCBS colonies could be identified as Vibrio spp. when compared with TCI (46%). SIGNIFICANCE: This study suggests that TCBS is a superior medium when compared with TCI for the isolation of Vibrio spp. from estuarine waters. Because of the public health risk presented by V. vulnificus, V. parahaemolyticus, V. cholerae and other vibrios, the selection of the most appropriate medium for their isolation is extremely important. PMID- 12581374 TI - Detection of Salmonella by indicator agar media and PCR as affected by alfalfa seed homogenates and native bacteria. AB - AIMS: To investigate and prevent the undesirable effect of native bacteria and alfalfa seed homogenates on detection of Salmonella in alfalfa seeds by indicator agar media and polymerase chain reaction (PCR). METHODS AND RESULTS: The relative sensitivity of five indicator agar media, including modified semisolid RV (MSRV), xylose-lysine-Tergitol 4 (XLT4), Hektoen enteric agar (HEA), brilliant green agar (BGA) and bismuth sulphite agar (BSA), for detection of Salmonella in the presence of a large number of native bacteria from alfalfa seeds was examined. The detection limit as measured by the ratio between the numbers of native bacteria and Salmonella was estimated to be 10(6) to 1 for MSRV and 10(3) to 1 for XLT4, HEA, BGA or BSA. Presence of alfalfa seed homogenates markedly reduced the sensitivity of Salmonella detection by PCR. The minimal number of Salmonella detectable by PCR was determined to be 1-10 and 100-1000 CFU in the absence and presence of seed homogenate, respectively. Application of anti-Salmonella immunomagnetic beads permitted detection of 2-5 CFU of heat-injured cells in 25 g of seeds within 24 h by PCR. CONCLUSIONS: The MSRV medium is more sensitive than other indicator agars for detecting a small number of motile Salmonella in samples containing a large number of native bacteria. Application of immunomagnetic beads eliminates the PCR-inhibitory activity of seed homogenates and improves the detection of Salmonella in inoculated seeds. SIGNIFICANCE AND IMPACT: The results generated from this study will aid the seed distributors, sprout growers and public health officials to identify and recall the Salmonella contaminated seed lots to be used for sprout production. PMID- 12581375 TI - Detection of the low-germination-rate resting oospores of Pythium myriotylum from soil by PCR. AB - AIMS: To establish a sensitive and specific polymerase chain reaction (PCR)-based method for detecting Pythium myriotylum in soils. METHODS AND RESULTS: Oospores of P. myriotylum were separated from large soil particles by flotation in sucrose solution. The thick-walled oospores were disrupted by vortex with sea sand and its DNA was extracted by the Cetyl trimethyl Ammonium Bromide (CTAB) method. The recovered DNA was verified by PCR amplification of a 150-bp target sequence of P. myriotylum. Samples of 10 g of soil were assayed; thus, the detection limit by PCR-based method was 10 oospores per gram soil. The method was successfully applied for the detection of P. myriotylum in soils collected in March, prior to planting of ginger crops. CONCLUSIONS: A PCR-based method for detecting P. myriotylum from soil was achieved. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR method has allowed us to monitor the presence of P. myriotylum in soil prior planting season as a way of reducing or eliminating disease. PMID- 12581377 TI - Growth and reduction of microorganisms in sediments collected from a greywater treatment system. AB - AIMS: To study the effects of competitive microbiota, temperature and nutrient availability on Salmonella, Enterococcus, Campylobacter spores of sulphite reducing anaerobes and bacteriophages MS2 and phiX174 in sediments from a greywater treatment system. METHODS AND RESULTS: Standard culture methods were used. Bacteria died off rapidly under normal conditions (20 degrees C, competitive microbiota) but remained stable or grew in the other conditions studied. When the sediments became nutrient depleted after 2 weeks, a log-linear die-off was observed for Salmonella, which was higher at 20 degrees C than at 4 degrees C. Bacteriophage decay was shown to be log-linear from day 0, with T90 values ranging from 9 (phiX174, 20 degrees C) to 55 days (phiX174, 4 degrees C). The MS2 phage had a significantly higher decay rate in tyndallized sediments (T90 = 17 days) than in original sediments (T90 = 47 days) (P < 0.001), with temperature not shown to affect the decay rate. Spores of sulphite-reducing anaerobes were not significantly reduced during the study period (35 days). Campylobacter died-off rapidly or entered a viable but non-culturable state and subsequently results were not provided. CONCLUSIONS: Competition was the most important factor to suppress pathogenic bacterial growth in an eutrophic environment. When nutrient depleted conditions prevailed, temperature was more important and log-linear decay of microorganisms could be observed. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings suggest that the normally occurring microbiota will suppress pathogenic bacterial growth in nutrient rich sediments. With lower nutrient status, temperature is the more important factor in reducing pathogens. PMID- 12581376 TI - Antibacterial activity of selected plant essential oils against Escherichia coli O157:H7. AB - AIMS: To quantify the antibacterial properties of five essential oils (EO) on a non-toxigenic strain of Escherichia coli O157:H7 in the presence and absence of a stabilizer and an emulsifier and at three different temperatures. METHODS AND RESULTS: Five EOs known to exhibit antibacterial properties were screened by disc diffusion assay and the most active were selected for further study in microdilution colorimetric assays. Oregano (Origanum vulgare) and thyme (Thymus vulgaris; light and red varieties) EO had the strongest bacteriostatic and bactericidal properties, followed by bay (Pimenta racemosa) and clove bud (Eugenia caryophyllata synonym: Syzygium aromaticum) EO. Oregano oil was colicidal at 625 microl l(-1) at 10, 20 and 37 degrees C. The addition of 0.05% (w/v) agar as stabilizer reinforced the antibacterial properties, particularly at 10 degrees C, whereas 0.25% (w/v) lecithin reduced antibacterial activity. Scanning electron micrographs showed extensive morphological changes to treated cells. CONCLUSIONS: Oregano and thyme EO possess significant in vitro colicidal and colistatic properties, which are exhibited in a broad temperature range and substantially improved by the addition of agar as stabilizer. Bay and clove bud EO are less active. Lecithin diminished antibacterial properties. The bactericidal concentration of oregano EO irreversibly damaged E. coli O157:H7 cells within 1 min. SIGNIFICANCE AND IMPACT OF THE STUDY: Oregano and light thyme EO, particularly when enhanced by agar stabilizer, may be effective in reducing the number or preventing the growth of E. coli O157:H7 in foods. PMID- 12581379 TI - Use of CAS-agar plate modified to study the effect of different variables on the siderophore production by Aspergillus. AB - AIMS: To evaluate the suitability of chrome azurol S (CAS) agar plate assay as a quantitative methodology for siderophore production. METHODS AND RESULTS: Aspergillus species (A. flavus, A. niger, A. tamarii) were inoculated in the CAS agar plates and the siderophores production was determined and expressed as CAS reaction rate (mm per day). All the species showed positive CAS reaction with different rates depending on culture conditions and A. flavus showed the highest CAS-reaction rate. The siderophore production in solid medium expressed as CAS reaction rate was correlated with siderophore production in liquid medium. CONCLUSIONS: The use of CAS-agar plate assay was modified and the evaluation of CAS reaction in mm per day made it possible to study and quantify the effect of several variables on the siderophore production by Aspergillus fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: We describe the CAS-agar plate assay as a quantitative methodology, which make it possible to select and evaluate the siderophore production by several microorganisms (fungi and bacteria) according to different culture conditions. PMID- 12581378 TI - Biodegradation of the synthetic pyrethroid cypermethrin in used sheep dip. AB - AIMS: To investigate the breakdown of cypermethrin synthetic pyrethroid (SP) insecticide-based used sheep dip (USD), with its indigenous microbial community and two previously isolated SP-degrading microorganisms. METHODS AND RESULTS: Cultures of USD (50 ml) containing 250 ml l(-1) cypermethrin were inoculated with the SP-degrading organisms and incubated at 25 degrees C with agitation at 80 rev min(-1) for 14 days. The viable cell counts and concentration of cypermethrin were monitored. A non-stimulated control was also carried out. The previously isolated bacteria were the most effective at degrading cypermethrin, leaving approximately two-thirds the concentration of SP as was found in the control. The non-stimulated cultures showed negligible breakdown of SP over the experimental period. CONCLUSIONS: The previously isolated SP-degrading bacteria could have a use in the treatment of SP USD. SIGNIFICANCE AND IMPACT OF STUDY: In situ treatment of SP-based USDs to detoxify the active ingredient before disposal could be very useful in helping to deal with agricultural pesticide waste. Such an approach, or by ex situ treatment would be more preferable to current methods, such as those of incineration and disposal to land. PMID- 12581380 TI - Production of gamma-linolenic acid by disrupted mycelia of Mortierella isabellina. AB - AIMS: To optimize the production of linolenic acid by disrupted mycelia of Mortierella isabellina. METHODS AND RESULTS: Effects of incubation conditions such as incubation time, pH of reaction mixture, concentration of Mg2+ or malate and incubation temperature on production of linolenic acid were studied. The production of gamma-linolenic acid reached 224 mg g-1 dry cells when the reaction mixture was composed of 1.0 g (dry mycelial mass) of disrupted mycelia of M. isabellina, 50 ml (50 mmol l(-1)) potassium phosphate buffer supplemented with 0.312 mmol l(-1) of Mg2+ and 10 mmol l(-1) of malate, pH 7.0 and incubated at 5 degrees C for 1 day. CONCLUSIONS: Incubation temperature, concentration of Mg2+ and malate showed major effects on the increased linolenic acid production. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights conditions for increasing gamma-linolenic acid production by cell-free mycelia of M. isabellina and an insight into rapidly gaining high production of polyunsaturated fatty acids. PMID- 12581381 TI - Identification of a novel insertion sequence in vanB2-containing Enterococcus faecium. AB - AIMS: The characterization of a novel insertion sequence (IS) in vanB2-containing Enterococcus faecium was conducted. METHODS AND RESULTS: Direct PCR amplification of ORFC region of Tn5382 from DNA extracted from vanB2-containing E. faecium, and sequence analysis were performed. A novel IS was identified. It is 1418 bp in length and contains one putative open reading frame that is similar to transposase. There exists inverted terminal repeats of 12 bp, but direct repeats are not present. According to high similarity to putative transposases of IS3 members, such as, IS150, IS861, IS1077 and IS911, we designated it ISEnfa3. SIGNIFICANCE AND IMPACT OF STUDY: Since ISEnfa3 was detected in all vanB2 containing strains examined so far, it could be used as a tool for epidemiological study. PMID- 12581382 TI - Cell death regulation in oral squamous cell carcinoma: methodological considerations and clinical significance. AB - In the last three decades, more work has been done on apoptosis and its role in the pathogenesis of many diseases including cancer. In almost all instances of cancer, dysregulation of cell death (apoptosis) and cell proliferation have been found to play a major role in tumourigenesis. A lot of progress has been made on understanding the molecular basis of apoptosis and its regulatory mechanisms. This review focuses on current knowledge on the regulation of apoptosis in oral squamous cell carcinoma, current methodologies and methodological consideration in estimation of cell death in tissue sections and the clinical significance of apoptosis related molecules in progression of oral squamous cell carcinoma. PMID- 12581383 TI - Transforming growth factor beta1 (TGFbeta1) expression in head and neck squamous cell carcinoma patients as related to prognosis. AB - BACKGROUND: Transforming growth factor beta1 (TGFbeta1) is a negative growth regulator in keratinocytes, and in vitro studies lead to the concept that loss of TGFbeta1 responsiveness is a critical step in epithelial carcinogenesis. OBJECTIVE: To investigate the prognostic relevance of TGFbeta1 expression in head and neck squamous cell carcinoma (HNSCC). MATERIALS AND METHODS: TGFbeta1 distribution was determined by immunohistochemistry in oral cavity/oropharynx (n = 79), larynx (n = 36) and hypopharynx (n = 25) tumors and in matched normal adjacent mucosa. TGFbeta-type I and II receptors were determined in 20 cases of differentiated oral cavity/hypopharynx tumors. Cases were considered positive if displaying reactivity in >10% of the cells. RESULTS: TGFbeta1-positive expression was found in 47.2% of larynx, 36.7% of oral cavity/oropharynx and in 24% of the hypopharynx tumors. Reactivity in >60% of the cells was displayed only by 11.4% of HNSCC. All normal controls were positive. TGFbeta1-positive expression did not correlate with clinico pathological parameters. An association with differentiation was verified only in oral cavity/oropharynx tumors (P or =1:400 was 100% (50/50), but its specificity only 55% (15/27) and the positive predictive value was 80% (50/62). Using a DAT titre > or =1:6400, the sensitivity was 84% (42/50), the specificity 85% (23/27) and the positive predictive value 91% (42/46). All four patients with DAT titre > or =1:6400 but negative splenic aspirate were also rK39 positive; we consider these are probably 'true' cases of kala-azar, i.e. false negative aspirates, rather than false DAT and rK39 seropositives. There were no false negative DATs (DAT titre < or =1:400 and aspirate positive), but there were four false negative rK39 tests (rK39 negative and aspirate positive). The rK39 dipstick is a good screening test for kala-azar; but further development is required before it can replace the DAT as a diagnostic test in endemic areas of the Sudan. PMID- 12581444 TI - The impact of adjustment for parity and mid-upper-arm circumference on sensitivity of symphysis-fundus height measurements to predict SGA foetuses in Mozambique. AB - BACKGROUND: With a new symphysis-fundus height (SFH) growth chart, based on Mozambican women with ultrasound-dated singleton pregnancies, the aim was to examine the possibility to enhance sensitivity of predicting small for gestational age (SGA) newborns by attempts to adjust the chart for parity and for mid-upper-arm circumference (MUAC). METHODS: Two antenatal clinics were chosen in the suburban area of Maputo City. A cohort of 904 consecutively recruited antenatal clients was followed until birth. Gestational age was determined by ultrasound at enrolment. The growth of the SFH was measured every 2-3 weeks. Women with multiple pregnancy or with gestational age >21 weeks at enrolment were excluded. Attempts were made to adjust SFH measurements for parity and MUAC by developing a mathematical model to increase sensitivity of the SFH method to predict a foetus being SGA. RESULTS: Parous women had on average 0.5-1 cm higher SFH readings than nulliparous women. Women with a body mass index (BMI) <19 and women with BMI >27 had approximately 1 cm lower and 1 cm higher readings, respectively, compared with women of normal BMI. There was a significant correlation between BMI and MUAC (r = 0.621; P < 0.001). The usefulness of SFH measurements to predict SGA newborns was analysed. The sensitivity was 49%, the specificity was 66%, the positive predictive value was 14% and the negative predictive value, 93%. By using the correlation between BMI and MUAC we tried to find a simple and useful method to improve the sensitivity of SFH to detect SGA foetuses. By reducing the SFH measurement by 1 cm for women with MUAC >29 and by 1 cm for multiparous women the sensitivity raised to 65% at the expense of reducing the specificity to 51%. Using a linear function of BMI, MUAC and parity to adjust the SFH measurement for each individual woman, it was possible to get a sensitivity of 70% with a corresponding specificity of 56%. CONCLUSION: By using BMI, MUAC and parity, it might be possible to improve the sensitivity of the SFH growth chart in predicting newborn being SGA but mostly at the expense of specificity. PMID- 12581445 TI - Longitudinal studies in South Indian villages on Japanese encephalitis virus infection in mosquitoes and seroconversion in goats. AB - Japanese encephalitis (JE) is endemic in Cuddalore district, Tamil Nadu, where Culex tritaeniorhynchus Giles was the major vector. We screened 45 100 adult female Cx. tritaeniorhynchus (902 pools) by enzyme-linked immunosorbent assay and isolated and confirmed JE virus (JEV) by using an insect bioassay system. We had 69 isolates of which 62 (90%) were identified as JEV. The average vector abundance per man hour for Cx. tritaeniorhynchus was 324.5 per month for the period June 1998-May 2000. The average minimum infection rate (MIR) per month in Cx. tritaeniorhynchus was 1.4 (range 0.0-5.6). Every year, a new batch of goats, 20 in the first year and 31 in the second year, born during the non-JE transmission period (January-June), aged <6 months and negative for haemagglutination inhibition (HI) antibodies were procured and placed in the villages as sentinels. Fortnightly, blood specimens were collected from these goats and tested for JE antibodies by HI test. Seroconversions (SCs) were recorded in 14 goats (70%) in the first year and 23 goats (74%) in the second year. JE HI antibody titres in goats were low (1:10-1:80) and these levels declined to undetectable levels in about 4 weeks following SCs. The time sequence of events indicated that four of five peaks of MIR in mosquitoes were followed 1 3 months later by peaks in the proportion of seroconverted goats. We suggest the screening of goats and cattle as a more feasible tool to stratify areas according to JE infection risk to the human population through the regular health system rather than screening mosquitoes using monoclonal antibodies, which is possible only in specialized laboratories. PMID- 12581446 TI - Prescribing practices of rural primary health care physicians in Uzbekistan. AB - Doctors prescribe medications for therapeutic indications and to meet patient expectations. Understanding the pattern of prescribing is a necessary precursor for any intervention and for improving prescribing practices. Using the WHO standard methodology, we investigated the prescribing practices of doctors in rural primary health care (PHC) clinics in the Ferghana region of Uzbekistan. Doctors in these clinics may have over-prescribed, prescribing 2.9 drugs per patient per encounter. Fifty-seven per cent of these were for injectable drugs, and 57% for antibiotics. Most prescriptions were for name brand (62%) rather than generic drugs. PMID- 12581447 TI - Human fascioliasis problem in a high-altitude area of Peru. PMID- 12581448 TI - An ABC for West Nile virus. PMID- 12581449 TI - Survey of autologous blood transfusion activity in England (2001). AB - Better Blood Transfusion' (Department of Health (1998) HSC 1998/224) stressed the need for National Health Service Trusts to explore the feasibility of autologous blood transfusions and raise patient awareness of these techniques. The development of an appropriate strategy to promote autologous transfusion requires an understanding of current use, along with constraints to increasing practice. Past surveys have shown unchanging attitudes to autologous transfusion (Torrella et al. (2001) Transfusion Medicine, 11, 15-19), but detailed information regarding the extent of autologous transfusion in England and North Wales was not available. This survey, carried out in 2001, considered autologous transfusion under four headings: predeposit autologous donation, acute normovolaemic haemodilution, intra-operative cell salvage and postoperative cell salvage. Questionnaires were returned from 265/310 (85.4%) hospitals served by the National Blood Service. Sixty per cent of respondents performed at least one autologous technique, although practice was limited to a few clinicians within each hospital. Predeposit donation was the most widespread technique but involved very few patients. Intra- and postoperative cell salvage techniques were practised in fewer hospitals but on many more patients. The main constraints to increasing use of autologous techniques were perceived to be logistical, but removal of constraints may only result in small increases in individual practice. More clinicians need to be encouraged to utilize appropriate, evidence-based autologous techniques. PMID- 12581450 TI - Transfusion medicine in India: a survey of current practice. AB - The new Indian National Blood Policy intends to improve the provision of easily accessible safe blood and blood components, available according to need. There is a requirement for information on patterns of blood transfusion in India, to help understand the potential for changes in transfusion practice and the organization of blood services, and to help ensure optimal use of this valuable and limited human resource. This study involved a questionnaire survey of blood transfusion practice with reference to Indian National AIDS Control Organisation clinical guidelines at selected blood centres in four study areas (Delhi, Bangalore, Nasik District and Imphal). Information on 1062 transfusion episodes was analysed. Adult recipients accounted for 87% of transfusions, and amongst the age group of 25-34, 73% of transfusions were to women. Anaemia was listed as a reason for 60% of transfusions, surgery for 42%, acute haemorrhage for 26% and pregnancy for 16%. Seventy-four per cent of adult transfusions were inappropriate when assessed against criteria derived from government transfusion guidelines. Possible common proximate causes for inappropriate transfusions include unnecessary transfusion for iron-deficiency anaemia and transfusion as a first choice for volume replacement. Options to promote good transfusion practice in India should be appraised. PMID- 12581451 TI - Quantification of feto-maternal haemorrhage (FMH) by flow cytometry: anti-fetal haemoglobin labelling potentially underestimates massive FMH in comparison to labelling with anti-D. AB - Many centres now routinely use flow cytometry to quantify feto-maternal haemorrhage (FMH). However, which flow cytometric method is the most accurate in quantifying FMH is currently unknown. An audit of clinical results in which FMH had been estimated by both directly conjugated monoclonal anti-D and anti-fetal haemoglobin (HbF) labelling suggested that the anti-HbF labelling method may underestimate massive FMH in comparison to labelling with anti-D. Subsequent to this audit, 46 samples of adult D-negative blood were spiked with varying amounts of D-positive cord blood (0.05-10% fetal cells per sample), and the number of fetal cells present was quantified by both labelling methods. The percentage of fetal cells detected by anti-D was not significantly different to the estimated percentage of fetal cells added to each sample (P = 0.636). However, anti-HbF labelling significantly underestimated the percentage of fetal cells present (P = 0.0001). In comparison to anti-D, the percentage of fetal cells detected by anti HbF was also significantly lower (P < 0.0001). The difference in fetal cell detection between anti-D and anti-HbF labelling was only apparent in the spiked samples containing > or =1% fetal cells per sample. In samples containing < or =0.6% fetal cells, no significant difference in the detection of fetal cells between anti-D and anti-HbF labelling was observed (P = 0.11). To allow adequate immunoprophylaxis in D-negative mothers with massive FMH, we recommend that anti D labelling should be used in the routine flow cytometric estimation of FMH. PMID- 12581452 TI - A preliminary comparison of flow cytometry and tube agglutination assays in detecting red blood cell-associated C3d. AB - This study compared flow cytometric analysis with tube agglutination assays for the detection of red blood cell (RBC)-associated complement and immunoglobulins (Igs). RBCs from 20 patients with reactive tube direct antiglobulin tests (DATs) were evaluated by flow cytometry with anti-C3d, anti-IgG, anti-IgM and anti-IgA. Serial samples were also tested from a patient at risk of passenger lymphocyte haemolysis. Results of flow cytometry and tube assays for anti-IgG were as follows: 12 of 20 samples reactive in both; six of 20 nonreactive in both; two of 20 discordant with a reactive tube and a nonreactive flow cytometry assay. Anti C3d results showed nine of 20 reactive in both and 11 of 20 discordant with a nonreactive tube and a reactive flow cytometry assay. In the IgM flow cytometry assay, three samples were reactive with anti-IgM. Samples from a group A woman who was transplanted with stem cells from a group B donor showed that on days 3 through 6 post-transplant, the flow cytometry assays for anti-IgG and/or anti-C3d were reactive, whilst the tube assays were nonreactive. In conclusion, flow cytometric analysis is more sensitive than the tube assay for the detection of RBC-associated C3d. Further studies are needed to determine the correlation of C3d levels with clinical sequelae. PMID- 12581454 TI - Alloimmune neonatal neutropenia linked to anti-HNA-4a. AB - This is a novel case report of alloimmune neonatal neutropenia (ANN) linked to the neutrophil antibody anti-HNA-4a (MART). Since its discovery, the HNA-4a antigen has never been associated with any clinical neutropenia. A first-born neonate with respiratory distress was found to be severely neutropenic, because of ANN. The broad reactivity of the antibody together with its capture by CD11b and CD18 in monoclonal antibody immobilization of granulocyte antigen test suggested HNA-4a specificity. DNA sequencing confirmed that the father is HNA-4a positive and that the mother is HNA-4a-negative, supporting the diagnosis of ANN linked to MART. PMID- 12581453 TI - A new murine monoclonal antibody against Kx protein. AB - Mice immunized with a synthetic peptide located on an intracellular segment of the polytopic Kx protein (37 kDa) from human red blood cells (RBCs) produced a monoclonal antibody called C7B8. As expected, this antibody did not agglutinate common RBCs but reacted with permeabilized cells in flow cytometry. C7B8 recognizes the Kx protein on Western blots. Cross-reactivity of C7B8 with human calpain of human muscle extracts was demonstrated by Western blot analysis. This cross-reactivity precludes the use of C7B8 for Kx tissue distribution studies, but immobilized C7B8 was a convenient tool for purification of the Kell-Kx complex from RBC membrane extract by immunochromatography. PMID- 12581455 TI - Anti-Ce complicating two consecutive pregnancies with increasing severity of haemolytic disease of the newborn. AB - The Ce antigen is expressed on red cells of individuals with Rh haplotypes, R1 or CDe and r' or Cde. However, anti-Ce is rare, and only two cases of severe haemolytic disease of the newborn (HDN) caused by this antibody have been described (Malde et al., 2000; Wagner et al., 2000). We describe a woman who was found to have anti-Ce in her second pregnancy, resulting in a neonate with HDN that required exchange transfusion. She subsequently had a twin pregnancy, where both the twins were affected by severe haemolytic disease (HD) of the fetus because of anti-Ce and required repeated fetal transfusions, followed by exchange transfusions after birth. This is the first reported case of HD caused by anti-Ce requiring fetal transfusions. PMID- 12581456 TI - Dieulafoy's lesion of duodenum: a case report. AB - BACKGROUND: Dieulafoy's lesion is an uncommon but important cause of recurrent upper gastrointestinal bleeding. Extragastric location of Dieulafoy's lesion is rare. We report two cases of Dieulafoy's lesion of the duodenum and discuss the management of this extremely uncommon entity. CASE PRESENTATION: Two cases of massive upper gastro-intestinal bleeding in young adults due to Dieulafoy's lesion of the duodenum are reported. Endoscopic diagnosis was possible in both cases. Hemostasis was achieved successfully by endoscopic adrenaline injection. The endoscopic appearance, pitfalls in the diagnosis and management of this rare lesion are discussed. CONCLUSIONS: Endoscopic diagnosis of extragastric Dieulafoy's lesion can be difficult because of the small size and obscure location of the lesion. Increased awareness and careful and early endoscopic evaluation following the bleeding episode are the key to accurate diagnosis. Adrenaline injection is one of the important endoscopic modalities for control of bleeding. PMID- 12581457 TI - Development and evaluation of a quality score for abstracts. AB - BACKGROUND: The evaluation of abstracts for scientific meetings has been shown to suffer from poor inter observer reliability. A measure was developed to assess the formal quality of abstract submissions in a standardized way. METHODS: Item selection was based on scoring systems for full reports, taking into account published guidelines for structured abstracts. Interrater agreement was examined using a random sample of submissions to the American Gastroenterological Association, stratified for research type (n = 100, 1992-1995). For construct validity, the association of formal quality with acceptance for presentation was examined. A questionnaire to expert reviewers evaluated sensibility items, such as ease of use and comprehensiveness. RESULTS: The index comprised 19 items. The summary quality scores showed good interrater agreement (intra class coefficient 0.60 - 0.81). Good abstract quality was associated with abstract acceptance for presentation at the meeting. The instrument was found to be acceptable by expert reviewers. CONCLUSION: A quality index was developed for the evaluation of scientific meeting abstracts which was shown to be reliable, valid and useful. PMID- 12581460 TI - Insights on food and nutrition in the Federated States of Micronesia: a review of the literature. AB - BACKGROUND: Nutrition-related disorders, including vitamin A deficiency (VAD) and chronic diseases, are serious problems in the Federated States of Micronesia (FSM). Many suggest that these disorders are new problems related to dietary and lifestyle changes. In the past four decades, imported foods, such as white rice, flour, sugar, refined foods and fatty meats, have increasingly replaced local foods in the diet. AIM: A literature review was conducted to understand underlying issues related to dietary change and obtain insights for nutrition research and interventions. METHOD: Published and unpublished papers from different disciplines were reviewed and collated for information on food and nutrition in FSM. Topics covered were historical background, local foods, infant and child feeding, dietary assessment, and nutritional status. Particular focus was on information and data relating to VAD, the primary topic that led to the review of the literature. CONCLUSIONS: FSM, a tropical country of abundant agricultural resources, has suffered a great loss in production and consumption of local foods. Inconsistent external and internal government policies and food aid programmes have contributed to the problem. Further research on the nutrient content of local foods and factors affecting production, acquisition and consumption is needed, as well as a broad, well-planned, intersectoral intervention aimed at dietary improvement for all age groups in the population. PMID- 12581461 TI - Relative efficiency of government and non-government organisations in implementing a nutrition intervention programme--a case study from Bangladesh. AB - OBJECTIVE: The Bangladesh Integrated Nutrition Programme (BINP) experimented with two models of delivery: the first model uses the Government of Bangladesh's (GOB) own management structure and the second uses the non-government organisations (NGOs) working in the local community. This study compares the relative efficiency of GOB and NGO management in the provision of nutrition services. DESIGN: A detailed costing survey was carried out to estimate the cost of delivering nutrition services from the Community Nutrition Centres (CNCs). The number of individuals enrolled, the number actually participating in the programme and person-days of service delivered were used as effectiveness measures. SETTING: Thirty-five CNCs were randomly selected from five BINP areas, of which 21 were in GOB-run areas and 14 in NGO-run areas. RESULTS: The cost of providing nutrition services per enrolee was US dollars 24.43 for GOB-run CNCs and US dollars 29.78 for NGO-run CNCs. CONCLUSIONS: Contrary to the widely held view, the analysis implies that the NGO facilities are not more efficient in the delivery of nutrition services when cost per person-days of service delivered is considered. The food cost component of BINP is so high that, irrespective of the delivery mode, policy makers should examine carefully the components of BINP in order to find the most cost-effective mix of services. PMID- 12581462 TI - Increased intake of fruit and vegetables: estimation of impact in terms of life expectancy and healthcare costs. AB - OBJECTIVES: There is strong evidence that a high consumption of fruit and vegetables reduces the risk of developing many cancers. This study examined the economic consequences for the healthcare sector if people followed the recommendations and increased their intake of fruit and vegetables. DESIGN: A life table was used to describe a base case population with respect to life expectancy, cancer incidence and healthcare costs. Relative risks of cancer for a high versus a low intake of fruit and vegetables were obtained from the literature and were used to simulate populations with a higher intake of fruit and vegetables. The empirical data consist of a 20% sample of the Danish population that was followed from 1993 to 1997. Civil registration numbers were used to link various computerised registers, in order to describe each individual in the sample in terms of morbidity, mortality and healthcare costs. RESULTS: The average daily intake of fruit and vegetables was assumed to be approximately 250 g for the general Danish population. Simulated intakes of 400 g and 500 g increased the life expectancy by 0.8 and 1.3 years, respectively. In addition, it was estimated that 19% and 32% of the cancer incidence could be prevented. The aggregate healthcare costs remained stable, as the resources saved due to a lower cancer incidence were offset by healthcare costs imposed by the fact that healthy people live longer and require more healthcare. However, the variations across age groups and health sectors were substantial. DISCUSSION: The study adopted a healthcare sector perspective. Only costs from hospitalisation and primary care were included in the calculations. The costs of changing people's dietary habits, i.e. education, information and promotion as well as other costs that would be relevant from a societal perspective, have not been taken into account. Furthermore, the transition from one level of intake to another is not the focus of the analysis, although it might take decades to observe the full effect of the dietary changes. CONCLUSION: Empirical evidence suggests that a considerable fraction of all cancer incidences can be prevented by a higher intake of fruit and vegetables. That may result in improved public health (gain in life years) at no additional cost to the healthcare sector. PMID- 12581463 TI - Achievement of dietary fatty acid intakes in long-term controlled intervention studies: approach and methodology. AB - OBJECTIVE: To describe the calculations and approaches used to design experimental diets of differing saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA) compositions for use in a long-term dietary intervention study, and to evaluate the degree to which the dietary targets were met. DESIGN, SETTING AND SUBJECTS: Fifty-one students living in a university hall of residence consumed a reference (SFA) diet for 8 weeks followed by either a moderate MUFA (MM) diet or a high MUFA (HM) diet for 16 weeks. The three diets were designed to differ only in their proportions of SFA and MUFA, while keeping total fat, polyunsaturated fatty acids (PUFA), trans-fatty acids, and the ratio of palmitic to stearic acid, and n-6 to n-3 PUFA, unchanged. RESULTS: Using habitual diet records and a standardised database for food fatty acid compositions, a sequential process of theoretical fat substitutions enabled suitable fat sources for use in the three diets to be identified, and experimental margarines for baking, spreading and the manufacture of snack foods to be designed. The dietary intervention was largely successful in achieving the fatty acid targets of the three diets, although unintended differences between the original target and the analysed fatty acid composition of the experimental margarines resulted in a lower than anticipated MUFA intake on the HM diet, and a lower ratio of palmitic to stearic acid compared with the reference or MM diet. CONCLUSIONS: This study has revealed important theoretical considerations that should be taken into account when designing diets of specific fatty acid composition, as well as practical issues of implementation. PMID- 12581464 TI - How well do children aged 5-7 years recall food eaten at school lunch? AB - OBJECTIVE: This study aimed to determine the accuracy with which children aged 5 to 7 years were able to report the food eaten at a school lunch. SUBJECTS/SETTING: Two hundred and three children (103 boys, 100 girls) aged 5-7 years were recruited from three primary schools in Oxford. DESIGN: Trained investigators made observational records of the school dinner and packed lunch intakes of four or five children per session. Children were interviewed within two hours of finishing the lunchtime meal and asked to provide a free recall of their meal. When the child had completed the recall, non-directive prompts were used to assess if the child was able to remember anything else. Foods recalled were classified as matches (recalled food agreed with observation), omissions (failed to report a food observed) or phantoms (recalled food was not observed). RESULTS: The percentage of accurate recall was significantly higher (P<0.01) in children eating packed lunch (mean 70 +/- 29%) than in children consuming school dinners (mean 58 +/- 27% ). This difference may have been due to increased familiarity of foods in packed lunches. Leftovers were not readily reported in this age group. Prompts and cues enhanced recall by all children. CONCLUSIONS: This study indicated that there was a wide range in the ability of children aged 5-7 years to recall intake from a packed lunch and/or school dinner. This dietary assessment method is unlikely to be suitable at an individual level. Investigators using dietary recall to estimate food intake in children aged 5-7 years need to be aware of the limitations of this method. PMID- 12581465 TI - Associates of obesity and weight dissatisfaction among Finnish adolescents. AB - OBJECTIVE: We investigated behavioural and socio-economic factors associated with obesity and weight dissatisfaction among Finnish adolescents. DESIGN: A total of 60,252 Finnish adolescents aged 14 to 16 years filled in a questionnaire about their health, health behaviour and socio-economic background. Food choices were obtained by using a short food-frequency questionnaire. Obesity was defined as a weight at least 120% of the sex- and height-specific mean weight for subjects. RESULTS: Of girls and boys, 54% and 66%, respectively, were satisfied with their weight. Among dissatisfied normal-weight adolescents, 81% of girls but only 48% of boys thought they were overweight. Of obese boys, 25% were satisfied with their weight. For both genders, obesity and weight dissatisfaction were associated with economic problems in the family. In girls, an association was also found with poor school performance, low educational level of parents and not having evening meals at home; and in boys, with physical inactivity and not eating school lunch. Smoking was more common among girls who were dissatisfied with their weight. Differences in food choices were small between different weight and weight satisfaction categories. CONCLUSIONS: Having normal weight and being satisfied with that weight are favourable for an adolescent. Obesity and weight dissatisfaction are associated mostly with disadvantageous health behaviours and low socio-economic status. Health behaviour seems to be associated more with weight satisfaction than with actual weight. PMID- 12581466 TI - Effect of a fruit and vegetable subscription in Danish schools. AB - OBJECTIVE: To measure the effect of a school fruit and vegetable subscription on children's intake of fruit and vegetables after 5 weeks of intervention. SETTING: Seven primary schools in Denmark. DESIGN AND METHODS: Intervention schools (n=4) were offered a fruit and vegetable subscription comprising one piece per day. Control schools situated in another municipality were not offered the subscription. Intake of fruit and vegetables was measured at baseline and 5 weeks after the start of the subscription. Two methods were used for dietary assessment: a pre-coded 24-hour recall form including total food intake and a food-frequency questionnaire (FFQ) including only fruit and vegetables. SUBJECTS: Children aged 6-10 years (n=804 from intervention schools and n=689 from control schools). Response rate in the dietary assessment was 31%. RESULTS: At intervention schools 45% of the children enrolled in the subscription. After 5 weeks of intervention, both subscribers and non-subscribers had increased their intake of fruit by 0.4 (P=0.019) and 0.3 (P=0.008) pieces per school day, respectively, but no change was observed in vegetable intake. Total intake increased only for non-subscribers by 0.4 piece/school day (P=0.008) mainly due to the consistent increase in fruit intake. No change in intake was measured at control schools. Only the 24-hour recall questionnaire was sensitive enough to pick up the changes of the subscription, whereas the FFQ was not. CONCLUSION: Five weeks with the subscription affected both subscribers and non-subscribers to increase intake of fruit. This may indicate that the subscription had an additional effect of stimulating parents of non-subscribers to supply their children with fruit. The results stress the importance of evaluating the effect of this type of programme, and the carefulness needed in designing the evaluation study. PMID- 12581467 TI - Size makes a difference. AB - OBJECTIVE: To elucidate status and trends in portion size of foods rich in fat and/or added sugars during the past decades, and to bring portion size into perspective in its role in obesity and dietary guidelines in Denmark. DATA SOURCES: Information about portion sizes of low-fat and full-fat food items was obtained from a 4-day weighed food record (Study 1). Trends in portion sizes of commercial foods were examined by gathering information from major food manufacturers and fast food chains (Study 2). Data on intakes and sales of sugar sweetened soft drinks and confectionery were obtained through nation-wide dietary surveys and official sales statistics (Study 3). RESULTS: Study 1: Subjects ate and drank significantly more when they chose low-fat food and meal items (milk used as a drink, sauce and sliced cold meat), compared with their counterparts who chose food and meal items with a higher fat content. As a result, almost the same amounts of energy and fat were consumed both ways, with the exception of sliced cold meat (energy and fat) and milk (fat). Study 2: Portion sizes of commercial energy-dense foods and beverages, and fast food meals rich in fat and/or added sugars, seem to have increased over time, and in particular in the last 10 years. Study 3: The development in portion sizes of commercial foods has been paralleled by a sharp increase of more than 50% in the sales of sugar sweetened soft drinks and confectionery like sweets, chocolate and ice creams since the 1970s. CONCLUSIONS: Larger portion sizes of foods low in fat and commercial energy-dense foods and beverages could be important factors in maintaining a high energy intake, causing over-consumption and enhancing the prevalence of obesity in the population. In light of this development, portion size ought to take central place in dietary guidelines and public campaigns. PMID- 12581468 TI - New Zealanders' attitudes to milk: implications for public health. AB - OBJECTIVE: To identify consumer attitudes and beliefs about (liquid) milk that may be barriers to consumption. DESIGN: Two random-quota telephone surveys conducted in Auckland one year apart. Respondents were questioned about their usual milk intake and their attitudes to milk. The questionnaire included attitude items that reflected the main themes of consumer interest in milk. SETTING: New Zealand. SUBJECTS: Seven hundred and thirteen respondents in the baseline survey and a separate sample of 719 respondents in the follow-up survey. RESULTS: At least one-third of the respondents consumed less than a glass (250 ml) of milk a day. Non-consumption was highest in young women (15%). People's concerns about milk related to what was important in their lives; what threatens them physically and emotionally. Women held more positive attitudes but they were concerned about the fat content of milk. Men were less aware of milk's nutritional benefits and as a result were less appreciative of its value. CONCLUSIONS: There is an opportunity to develop public health initiatives to address the barriers to drinking milk. Industry-health alliances may be an effective means to provide positive nutrition messages about milk and to engage the support of health professionals. PMID- 12581469 TI - Calibration of the dietary questionnaire for the Canadian Study of Diet, Lifestyle and Health cohort. AB - OBJECTIVE: For proper interpretation of results from epidemiological studies that use food-frequency questionnaires (FFQs), it is necessary to know the relationship between reported intakes from the FFQ and true usual intake. In this paper, we report a calibration study conducted to investigate the performance of the FFQ used in a cohort study, the Canadian Study of Diet, Lifestyle and Health. METHODS: Over a 1-year period, 151 men and 159 women completed a full set of questionnaires including a self-administered baseline FFQ, three 24-hour diet recalls administered by telephone, and a second FFQ self-administered subsequently. The association between the nutrient estimates derived from the FFQs and the diet recalls was evaluated by calculating deattenuated Pearson's correlation coefficients. RESULTS: FFQs estimated mean daily nutrient intakes higher than the diet recalls. When the log-transformed and energy-adjusted nutrient intakes from the average of three 24-hour recalls were compared against the baseline FFQ, the following deattenuated correlations were obtained in men and women, respectively: total energy 0.44 and 0.32, total fat 0.64 and 0.68, saturated fat 0.68 and 0.70, dietary fibre 0.65 and 0.44, vitamin E 0.32 and 0.37, vitamin C 0.40 and 0.37, beta-carotene 0.34 and 0.29, alcohol 0.74 and 0.67, caffeine 0.81 and 0.76, with a median correlation of 0.49 and 0.53. Correlations between the second FFQ and diet recalls were similar. The correlations between the two FFQs as a test of reliability had a median value 0.64 for men and 0.63 for women for selected nutrients. CONCLUSIONS: The study suggests that the FFQ method gives acceptable levels of nutrients or food component estimates, as assessed by this calibration study against diet recalls, when limited to energy-adjusted and deattenuated values. PMID- 12581470 TI - Complex correlation structure in consumption rates of major food groups: implications for the assessment of radiation exposure. AB - OBJECTIVE: An extensive dataset on individual food consumption was analysed in order to study all pairwise correlations between the consumption rates of 11 major food groups. Additionally, the project aimed to examine and quantify the accuracy of a recently proposed estimator of total food consumption to be used for the estimation of radiation exposure by food. Such an inquiry seems justified, because the proposed estimator implicitly presumes an essentially positive correlation structure in food consumption rates. DESIGN: Statistical analysis using representative data gathered in Germany in a nation-wide food consumption survey. SETTING: Germany. SUBJECTS: Individuals aged between 4 and 94 years namely 10,901 males and 12,308 females. RESULTS: The consumption rates of 11 major food categories showed several significantly positive, but also a number of significantly negative, correlations. Negative associations between cereal and potato products persisted consistently over all age groups, independent of sex. Other significantly negative correlations were limited to certain age groups. Reflecting these negative correlations, a subsequent analysis of relative ranks of consumption revealed that no person in the sample had the highest consumption rates in all food groups simultaneously. Based on representative samples, overestimations of 34 to 53% were obtained if--as recently suggested in the context of radiation exposure prediction--the 95th percentiles of total food consumption were determined as sums of the corresponding percentiles of the food groups. CONCLUSIONS: The complex correlation structure of food group consumption rates, as identified in this study, bears important implications for various health-related issues. Ignoring them could lead to overly conservative estimations of radiation exposure due to food ingestion or to confounding effects in epidemiological studies on nutritional risk factors of diseases. The results also indicate that a distinction into different dietary patterns might be useful in characterising different consumption habits. PMID- 12581471 TI - Urinary iodine excretion in pregnant women residing in areas with adequate iodine intake. AB - OBJECTIVE: Pregnancy is accompanied by profound alterations in thyroid economy and relative iodine deficiency. The aim of this study was to evaluate urinary iodine excretion of pregnant women in cities with adequate and more than adequate iodine intake. METHODS: A cross-sectional study was performed on schoolchildren and pregnant women in four cities in the Islamic Republic of Iran. Urinary iodine excretion was measured for 438 schoolchildren and 403 pregnant women. In addition, in Isfahan City, thyroid volume was measured by sonography for 30 pregnant women in each trimester of pregnancy and for 90 non-pregnant women who also had urinary iodine measurement. RESULTS: Median urinary iodine of schoolchildren was 31.2, 25.0, 20.2 and 19.3 microg/dl in Rasht, Isfahan, Ilam and Tehran, respectively. Corresponding values for pregnant women were 33.8, 21.2, 19.0 and 18.6 microg/dl. The percentage of pregnant women with urinary iodine below 20 microg/dl was 16, 45, 55 and 54, and below 10 microg/dl was 1, 7, 7 and 13, in Rasht, Isfahan, Ilam and Tehran, respectively. In Isfahan, urinary iodine was significantly decreased in the third trimester of pregnancy, compared with controls. Mean thyroid volume was 7.8 +/- 3.1 ml and 7.8 +/- 2.8 ml in pregnant and non-pregnant women, respectively. CONCLUSION: Recommended values for dietary iodine through universal salt iodisation may not be adequate for pregnant women, and the specific problem of iodine and pregnancy should be considered further in the light of the latest recommendations. PMID- 12581472 TI - Department of Health Research Initiative on Nutrition Phase 2: overview of findings and recommendations. AB - OBJECTIVE: The purpose of this overview is to synthesise and summarise the findings and recommendations of all 14 projects funded between 1997 and 2002 under Phase 2 of the Department of Health's Policy Research Programme Nutrition Initiative. This is aimed at end users, including policy makers, practitioners, researchers and research funding bodies. DESIGN: The main findings and recommendations for policy and practice, as well as for future research, contained in the peer-reviewed final report of each project are summarised under headings identified as being useful for policy groups and practitioners: Maintenance of a healthy weight; Functional outcomes related to different markers of iron status; Nutritional aspects of bone health in humans; and Dietary interventions. RESULTS: The overview draws together the key findings and recommendations for current and future policy and practice from the second phase of the research programme and identifies research gaps. CONCLUSIONS: The findings and recommendations of the Department of Health's Nutrition Research Initiative have contributed to the scientific evidence base for policy development, policy evaluation, and will inform practitioners as well as researchers and research funding bodies. PMID- 12581474 TI - The effectiveness of constraint induced movement therapy in two young children with hemiplegia. AB - Constraint induced movement therapy (CIMT) for hemiplegia involves constraining use of the unaffected limb while providing intensive shaping and practice of movements in the hemiplegic limb. The technique had been shown to be highly effective in improving upper limb function in adults following stroke, but there is only a limited literature on the use of this intervention in children. This paper provides a brief overview of the theory and background of this procedure, and reviews the literature on use of the technique in children. It then provides detailed case reports for two hemiplegic children, ages 19 and 38 months, each of whom underwent a trial of CIMT. Both children made significant gains in upper arm function that were reflected in a variety of domains, including aspects of everyday functional limb use. Gains persisted to variable degrees and some unexpected new gains were noted following cessation of CIMT. Practical challenges for the children, parents, and therapists in implementing this intensive but promising intervention are also discussed. PMID- 12581475 TI - Re-admissions to inpatient paediatric pulmonary rehabilitation. AB - OBJECTIVE: To describe re-admission rates, identify reasons for re-admission and examine characteristics of children requiring re-admission to inpatient pulmonary rehabilitation. METHODOLOGY: Retrospective record review of infants and toddlers (less than three years of age) requiring oxygen or ventilator support discharged from an inpatient paediatric pulmonary rehabilitation programme between 1992 and 1999. RESULTS: Forty-one initial admissions resulted in 45 readmissions with a mean re-admission rate of 1.1 (SD = 1.41) re-admissions per child. Children with re-admissions (n = 22, 54%) required significantly more ventilator support (p = 0.001) and nursing care (p = 0.001) and were transferred to acute care more frequently (p = 0.002) than children without re-admissions. One-half of the children re-admitted to inpatient pulmonary rehabilitation were re-admitted two or more times. CONCLUSIONS: Based on this cohort of children, dependence on supplemental oxygen and/or mechanical ventilation and medical complexity may be indicators that children will require re-admission to rehabilitation following a transfer back to acute care. Further examination of re-admission rates and reasons and children's clinical characteristics may have predictive value and provide practice improvement opportunities. PMID- 12581476 TI - Benefits of a virtual play rehabilitation environment for children with cerebral palsy on perceptions of self-efficacy: a pilot study. AB - This paper presents the results of a clinical trial of a virtual reality play based intervention. The results of three single case studies are presented. The virtual reality intervention used the Mandala Gesture Xtreme technology. It was applied to three school-aged children with cerebral palsy. A pre-test-post-test design was used. The relevant outcome of interest was self-efficacy as measured with the Canadian Occupational Performance Measure (COPM). Group scores on the COPM indicated clinically significant changes in self-efficacy for all children. Two of the participants demonstrated the greatest changes in both perceived performance abilities and satisfaction with performance with respect to task specific domain areas. Qualitative comments from the participants revealed a high degree of motivation, interest, pleasure, and opportunity for engagement in play, activities not previously engaged in. Overall, this pilot study suggests the viability of a virtual reality play-based intervention as part of the rehabilitation process for children with cerebral palsy. These results form the basis of a larger scale randomized clinical trial. PMID- 12581477 TI - The assessment of parent needs following paediatric traumatic brain injury. AB - The paediatric version of the Family Needs Questionnaire was administered to parents of children with traumatic brain injuries (TBI) a year or more post injury. Parents of diabetic children (DIAB) and parents of orthopaedically (ORTHO) injured children were included as control groups. Parents rated items on their importance and how well these needs have been met. The ORTHO group rated significantly fewer items as important relative to the TBI and DIAB groups (who endorsed a similar number of items). Of those items rated as important, TBI parents' needs were more likely to remain unmet relative to both the DIAB and the ORTHO groups: out of 28 items rated as important, TBI parents report that 19 needs are still unmet. DIAB and ORTHO parents more consistently reported their needs as met. Needs most often rated as unmet for TBI parents include the needs for health/medical information, professional support, community support networks, and the need to be involved in their child's care. These results present important findings for TBI rehabilitation professionals. Future studies need to investigate whether TBI parents' needs are unmet because of the lack of community resources, or if available resources do not adequately target the needs of the TBI parents. PMID- 12581478 TI - Aerobic capacity in late adolescents infected with HIV and controls. AB - OBJECTIVES: The objective of this study was to determine if aerobic capacity was diminished in late adolescents infected with HIV compared to controls matched for age, gender, and physical activity level. STUDY DESIGN: This study was a quasi experimental cross sectional analysis. Subjects (11 female, four male) were 15 late adolescents with HIV (18 +/- 0.03 years) (CD4: 499.2 +/- 37.5 cells/mm(3), viral load: 22043 +/- 9976.6 copies/ml, haematocrit: 36.4 +/- 1.2) and 15 age, gender, and activity level matched controls (18 +/- 0.03) who underwent maximal treadmill exercise testing, while oxygen consumption, carbon dioxide production, ventilation, and ECG data were simultaneously recorded via open circuit spirometry and electrocardiography. RESULTS: Peak oxygen consumption (p < 0.003), peak treadmill stage (p < 0.003), treadmill duration (p < 0.004), and oxygen pulse (p < 0.009) were lower in those infected with HIV compared to controls. Functional aerobic impairment was observed in the late adolescents infected with HIV, pointing toward pathological limitations of the oxidative metabolic pathway. CONCLUSION: This study has demonstrated that aerobic capacity was reduced substantially in late adolescents infected with HIV, below that observed in controls. The findings suggest that this decrease in oxidative capacity was due to mechanisms other than physiologic deconditioning. PMID- 12581479 TI - Microbiology and management of post-surgical wounds infection in children. AB - The recent increased recovery of anaerobic bacteria from children has led to greater appreciation of their role in paediatric infections at all body sites, including post-surgical wounds (PSW). In studies that employed adequate method for recovery of aerobic and anaerobic bacteria polymicrobial, aerobic and anaerobes were isolated from over half of the patients with PSW. The wounds studied were those that developed following these surgical procedures: head and neck surgery for malignancies, post-thoracotomy, spinal fusion and gastrostomy tube insertion. Staphylococcus aureus and aerobic gram-negative bacilli were found at all sites. However, a correlation was generally found between the site of the wounds and microbial flora recovered from the wound. Organisms that resided in the mucous membranes close to the surgical site predominated in the wound next to these areas. Enteric Gram-negative rods, Group D enterococcus and Bacteroides fragilis group predominated in wounds relating to the gut flora, while Streptococcus spp., pigmented Prevotella and Porphyromonas spp. and Fusobacterium spp. were most frequently recovered in wounds proximal to the oral area. Management of PSW should include administration of antimicrobials effective against the polymicrobial bacterial flora adjacent to the anatomic site of the wound. PMID- 12581481 TI - Detection of DNA strand breakage in a marine amphipod by agarose gel electrophoresis: exposure to X-rays and copper. AB - This article describes the leading steps to develop an assay of DNA damage for the marine amphipod Gammarus locusta, using agarose gel electrophoresis (AGE). To test the sensitivity and feasibility of the AGE technique, X-ray assays were performed with naked DNA and with live amphipods. These positive controls demonstrated the effectiveness of the AGE technique to not only discriminate distinct levels of DNA strand breakage in a dose-dependent manner, but also to identify and quantify the type of strand breakage induced. It was also shown that it is possible to detect DNA damage using whole-body DNA extracts from amphipods. To explore the potential of this technique for use in ecotoxicological studies with amphipods, a 96-h waterborne-copper toxicity test was performed. Copper induced DNA strand breakage was first observed after 24 h of exposure, and was recorded again at 96 h, at a copper concentration of 20 microg l(-1). The absence of strand breakage after 48 h of exposure is discussed in the light of the underlying mechanisms of copper toxicity and DNA repair. These studies demonstrated the feasibility of including DNA damage as a biomarker in ecotoxicological studies with amphipods. Information gained from the use of this biomarker would help with the interpretation of chronic toxicity tests and would contribute to our understanding of the impact of genotoxic insult in marine invertebrates, particularly crustaceans. PMID- 12581480 TI - Polymorphism of metallothionein genes in the Pacific oyster Crassostrea gigas as a biomarker of response to metal exposure. AB - Quantification of metallothioneins (MTs) is classically associated with a cellular response to heavy metal contamination and is used in the monitoring of disturbed ecosystems. Despite the characterization of several MT genes in marine bivalves, only a few genetic studies have used MT genes as potential biomarkers of pollution. The aim of this study was to assess whether MT gene polymorphism could be used to monitor exposure of the Pacific oyster Crassostrea gigas to heavy metals and to develop specific genetic markers for population genetic studies in relation to environmental stress. The polymorphism of two exons of the C. gigas MT gene CgMT1 were studied using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) in both field populations exposed to various metals concentrations and in experimentally exposed populations. High frequencies of two SSCP types in exons 2 and 3 of the CgMT1 gene have found to be significantly associated with tolerance to metals in experimental and field oyster populations. The use of MT1 gene polymorphism in C. gigas as in the present study should therefore be of high ecological relevance. In conclusion, the analysis of the types in these two CgMT1 gene exons, which can confer a greater tolerance to heavy metals, can constitute a good biomarker of effect of the presence of heavy metals in ecosystems. PMID- 12581482 TI - Changes in serum pneumoproteins caused by short-term exposures to nitrogen trichloride in indoor chlorinated swimming pools. AB - Nitrogen trichloride (NCl(3)) is an irritant gas released in the air of indoor pools sanitized with chlorine-based disinfectants. In the present study we investigated the effects of NCl(3) on the pulmonary epithelium of pool attendees by measuring the leakage into serum of three lung-specific proteins (pneumoproteins): the alveolar surfactant-associated proteins A and B (SP-A and SP-B) and the bronchiolar 16 kDa Clara cell protein (CC16). These pneumoproteins were measured in the serum of 29 recreational swimmers (16 children and 13 adults) before and after attending a chlorinated pool with a mean NCl(3) concentration of 490 microg m(-3). Pneumoprotein changes in serum were also studied in 14 trained swimmers performing an intensive 45 min standardized swimming session in a chlorinated pool (mean NCl(3) concentration of 355 microg m(-3)) and for the purposes of comparison in a non-chlorinated pool sanitized by the copper/silver method. Serum CC16 was not increased in recreational swimmers, but in trained swimmers serum levels of this protein peaked immediately after strenuous exercise, both in the copper/silver pool and in the chlorinated pool. This acute increase in airway permeability is probably the consequence of the mechanical stress on the epithelial barrier caused by overinflation and/or hyperventilation during intense exercise. Serum levels of SP-A and SP-B were unaffected by strenuous exercise in the copper/silver pool. The two proteins were, however, significantly increased in a time-dependent manner in recreational and trained swimmers attending the chlorinated pool. The intravascular leakage of SP-A and SP-B was already statistically significant after only 1 h of exposure to pool air without exercising and remained elevated for 12 h after. These changes were not associated with decrements in lung function. The ability of NCl(3) to acutely disrupt the lung epithelium barrier was confirmed in mice using serum CC16 and plasma proteins in bronchoalveolar lavage fluid as permeability markers. The significance of these permeability changes induced by NCl(3) in the deep lung is presently unknown. In view of the increasing and widespread human exposure to this gas not only in indoor pools but also in a variety of other situations, these findings warrant further study. PMID- 12581483 TI - Metallothionein concentration in the mussel Mytilus galloprovincialis as a biomarker of response to metal contamination: validation in the field. AB - Mussels were translocated from a shell-fish breeding area (Sete, on the French Mediterranean coast) to sites exposed to trace element inputs in April 2000. They were recovered 3 months later. Whole soft tissues from all of the sites (n = 97) were analysed for arsenic, cadmium, chromium, copper, mercury, nickel, lead and zinc. Metallothioneins (MTs) were also measured in the digestive gland and in the remaining tissues (allowing calculation of whole soft tissue concentrations) at 22 of the 97 sites. MT concentrations in the digestive gland and the whole soft tissues were strongly correlated. The condition index varied with food availability at different sites. This did not influenced MT concentrations in the whole soft tissues, whereas the condition index was negatively correlated to trace element concentrations. A model is proposed to minimize this influence of condition. Metal concentrations adjusted using this model showed significant correlations with MT levels for those metals (cadmium, copper, nickel and zinc) that are known to bind to this protein, with the exception of mercury. Even in moderately contaminated sites, measurement of the MT level in the soft tissues of mussels was generally able to discriminate between different levels of contamination, allowing the use of a simplified procedure compared with dissection of the digestive gland. It is recommended to avoid translocation and sampling during the reproductive period, which is well documented for commercial species such as Mytilus sp. PMID- 12581487 TI - Ingested type I interferon: a potential treatment for autoimmunity. AB - We have proposed a unifying hypothesis of the etiopathogenesis of autoimmunity that defines autoimmunity as a type I interferon (IFN) immunodeficiency syndrome. We have examined toxicity and potential efficacy in three phase I (type 1 diabetes, rheumatoid arthritis, multiple sclerosis) and one phase II clinical trials in multiple sclerosis (MS). In a phase I open-label trial in type 1 diabetes, ingested IFN-alpha preserved residual beta cell function in recent onset patients. In a second phase I trial, treatment of rheumatoid arthritis (RA) with ingested IFN-alpha reduced the secretion of interleukin-1 (IL-1), a proinflammatory cytokine. In a third phase I trial in MS, there was a significant decrease in peripheral blood mononuclear cell (PBMC) IL-2 and IFN-gamma production after ingesting IFN-alpha. In a phase II randomized, placebo controlled, double-blind trial in MS, 10,000 IU ingested IFN-alpha significantly decreased gadolinium enhancements compared with the placebo group at month 5. Tumor necrosis factor-alpha (TNF-alpha) and IFN-gamma cytokine secretion in the 10,000 IU group at month 5 showed a significant decrease that corresponded with the effect of ingested IFN-alpha on decreasing gadolinium enhancements. Ingested IFN-alpha was not toxic in any of these clinical trials. These studies suggest that ingested IFN-alpha may have a potential role in the treatment of autoimmunity. PMID- 12581488 TI - IFN-alpha2b increases interleukin-10 expression in primary activated human CD8+ T cells. AB - Interleukin-10 (IL-10) is a multifunctional cytokine with diverse effects on most hematopoietic cell types. It appears the principal function of IL-10 is to limit and ultimately terminate inflammatory response. We demonstrate here that interferon-alpha2b (IFN-alpha) increases the expression of IL-10 in activated primary CD8(+) T cells. Optimal induction of mRNA expression and protein synthesis was observed when IFN-alpha was added to cells activated by the combination of anti-CD3 monoclonal antibody (mAb) and IL-2. Maximal stimulation of IL-10 protein production was observed after prolonged incubation periods (48 72 h). No effects were observed on the production of IL-4, whereas IFN-gamma was produced with a faster kinetics than an untreated control. Our data indicate that IFN-alpha promotes the development of a CD8(+) T cell population with enhanced anti-inflammatory activity, which may play a critical role in the regulation of a proper immune response. PMID- 12581489 TI - Increased IFN-gamma protein in bronchoalveolar lavage fluid of anti-IP-10 antibody-treated F344 rats following Sendai viral infection. AB - A previous study showed that virus-resistant F344 rats had higher levels of interferon-gamma (IFN-gamma)-inducible protein 10 (IP-10) than did virus susceptible BN rats early after Sendai viral infection. The initial goal of this study was to determine if an early high expression of IP-10 in F344 rats contributes to their resistance to virus-induced airway injury. Infected F344 rats were treated with anti-IP-10 rabbit serum or normal rabbit serum. Results indicated that blocking of IP-10 protein did not significantly change the resistance of F344 rats. However, we observed that neutralization of IP-10 increased IFN-gamma protein in bronchoalveolar lavage (BAL) fluid of F344 rats 7 days after inoculation compared with rats that received normal rabbit serum. The pulmonary IFN-gamma mRNA abundance remained comparable. This effect was not caused by fluctuation of the viral titer in the lung. This interesting phenomenon suggests that expression of IFN-gamma protein can be modulated by treatment with anti-IP-10 antibody at the posttranscriptional or translational level in this model. PMID- 12581490 TI - A comparison of the biologic activity of two recombinant IFN-beta preparations used in the treatment of relapsing-remitting multiple sclerosis. AB - Recent clinical trials with interferon-beta (IFN-beta) in relapsing-remitting multiple sclerosis (RRMS) have clearly demonstrated that the IFN-beta dosing regimen affects the clinical efficacy, thereby highlighting the importance of determining the relative biologic activities of the IFN-beta products currently available. Although studies have been published that examine the biologic activities of the two structurally different forms of recombinant IFN-beta, IFN beta1a (Rebif), Serono, Geneva, Switzerland) and IFN-beta1b (Betaseron)/Betaferon), Berlex [Montville, NJ]/Schering [Berlin, Germany]), there have been few direct comparative studies. Therefore, to obtain a more accurate estimate of the relative biologic activities of Rebif and Betaseron, this study examined the antiviral activities of these two products within the same assay system and against the same natural human IFN-beta standard. Whereas the manufacturers' information suggests that the bioactivity of Betaseron is only about 8.7-fold less than that of Rebif, the results of the present direct, comparative study show that Rebif has an antiviral activity 14 times greater than that of Betaseron. This may have important clinical implications, because on the basis of the results reported here, Rebif at 44 microg t.i.w. is approximately double the maximal licensed weekly dose for Betaseron. PMID- 12581484 TI - Autometallography and metallothionein immunohistochemistry in hepatocytes of turbot (Scophthalmus maximus L.) after exposure to cadmium and depuration treatment. AB - In this study, autometallography and immunohistochemistry were used to localize and quantify cadmium and metallothionein (MT) levels, respectively, in cellular compartments of turbot liver on exposure to cadmium for 7 days and further depuration treatment for 14 days. Metals weakly bound to proteins (i.e. MTs) in hepatocyte lysosomes were visualized as black silver deposits (BSDs) using a light microscope. With the aid of a newly developed immunohistochemical procedure, MTs were localized and semi-quantified in both the cytosolic and the lysosomal compartments of hepatocytes. The BSD extent in the lysosomes of hepatocytes increased significantly as a result of cadmium exposure. This response was evidenced after 1 h. Further, a progressive increase in the volume density of BSDs occurred up to the seventh day. Total MT immunohistochemical levels increased at a lower rate, starting after 1 day of cadmium exposure. BSD extent values recovered after depuration, whilst MT levels remain unchanged. It is possible that the detoxification rate of metals via lysosomes was diminished, whilst MT levels remained unchanged, at least after 14 days of depuration. It can be concluded that autometallography and MT immunohistochemistry are good tools for clarifying metal and metal-MT trafficking routes in hepatocytes, and also that BSD extent and MT immunohistochemical levels in the lysosomes and cytosol of fish hepatocytes can be considered to be useful biomarkers of metal exposure. PMID- 12581485 TI - Toxicity of cypermethrin: hsp70 as a biomarker of response in transgenic Drosophila. AB - Heat shock protein induction is often associated with a cellular response to a harmful environment or to adverse life conditions. The main aims of our study were (1) to evaluate the cytotoxic potential of cypermethrin; and (2) to investigate the suitability of stress-induced heat shock protein Hsp70 as a biomarker for environmental pollutants in transgenic Drosophila melanogaster (Hsp70-lacZ)Bg(9). Different concentrations of cypermethrin (0.002, 0.2, 0.5 and 50.0 p.p.m.) were mixed with food. Third instar larvae of transgenic Drosophila melanogaster were allowed to feed on these mixtures for different time intervals (2, 4, 6, 12, 24 and 48 h). Following feeding, hsp70 induction and tissue damage were evaluated. In the highest concentration treatment group (50 p.p.m.), 100% larval mortality was recorded after 12 h exposure. Hsp70 was found to be induced even at the lowest concentration (0.002 p.p.m.) of the insecticide, while tissue damage was observed in the larvae exposed for 48 h. While an insignificant decline in hsp70 expression was observed in the larvae exposed to cypermethrin at a dietary concentration of 0.002 p.p.m. after 48 h compared with those exposed for 24 h, in the next two higher concentrations of the toxicant, a similar but significant decline in hsp70 expression was evident in the exposed larvae after 48 h. The present study reveals the cytotoxic potential of cypermethrin and further proposes that hsp70 induction in transgenic Drosophila melanogaster could be used as a sensitive biomarker in risk assessment. PMID- 12581491 TI - Thrombopoietin production in human hepatic cell cultures (HepG2) is resistant to IFN-alpha, IFN-beta, and IFN-gamma treatment. AB - Thrombocytopenia is an important complication of interferon (IFN) therapy for chronic viral hepatitis. To study whether IFN interferes with hepatic thrombopoietin (TPO) synthesis, we used the human hepatoma cell line HepG2. Our results show that IFN-alpha, IFN-beta, or IFN-gamma did not impair TPO mRNA expression, as determined by quantitative RT-PCR, even when high IFN doses (up to 5000 U/ml) or long-term incubations (up to 14 days) were applied. Neither was the rate of secretion of immunoreactive TPO reduced on IFN treatment. These findings support the concept that IFNs primarily mediate effects on megakaryocytic cells and platelets rather than on TPO-producing hepatocytes. PMID- 12581493 TI - Immune-specific immunoglobulin G-mediated enhancement of human immunodeficiency virus-induced IFN-alpha production. AB - Interferon-alpha (IFN-alpha) is synthesized as an integral part of innate immunity to viral infection. We previously provided preliminary evidence that antibody-containing serum from HIV-infected individuals enhanced HIV-induced production of IFN-alpha. Subsequently, preparations of pooled human immunoglobulin G (IgG) have also been shown to enhance poliovirus (PV)-induced IFN-alpha production. The current work establishes IgG as the serum mediator that enhances induction of IFN-alpha by HIV. Our studies also establish the ability of sera from individual subjects to enhance PV-induced IFN-alpha production. HIV induced IFN-alpha production was enhanced maximally by >4000-fold and by an average of 25-fold. Sera from 74 people enhanced PV- induced IFN-alpha from undetectable levels to an average of 615 units (range 7-4679 units). The ability of individual sera to enhance IFN-alpha production by HIV and PV persisted undiminished in patients with AIDS. IgG-mediated enhancement of IFN-alpha production was similar to that induced by IgG and PV and was blocked by IgG Fc fragments. Demonstration of the selective enhancement of HIV-induced IFN-alpha production by IgG from HIV-seropositive individuals provides further evidence for the existence of antigen-specific upregulation of a critical component of innate antiviral immunity by the adaptive Th2 immune response. PMID- 12581492 TI - Impaired macrophage responses may contribute to exacerbation of blood-stage Plasmodium chabaudi chabaudi malaria in interleukin-12-deficient mice. AB - Aiming to clarify the role of endogenous interleukin-12 (IL-12) in protective immunity against blood stages of Plasmodium chabaudi chabaudi (AS), we evaluated the course of infection in IL-12p40 gene knockout (IL-12p40KO) and wild-type (WT) C57BL/6 mice, focusing (1) on the ability of T cells to develop adequate type 1 responses and (2) on the potentiality of macrophages to respond to parasites, interferon-gamma (IFN-gamma), or both. We observed that IL-12p40KO mice develop significantly higher parasitemias during the acute infection, although mice from both groups clear the parasites within a month and similarly eliminate a secondary challenge. Thus, fully protective immunity to P. c. chabaudi can be generated in the absence of IL-12. However, this cytokine may promote parasite control during the early phase of infection. The increased acute parasitemia of IL-12p40KO mice was associated with both impaired IFN-gamma and nitric oxide (NO) response by spleen cells. Because stimulation with recombinant IFN-gamma (rIFN gamma) failed to improve the NO response in IL-12p40KO macrophages, we investigated whether these cells have an intrinsic defect. Analysis of peritoneal macrophages revealed that IL-12p40KO cells produce higher levels of transforming growth factor-beta1 (TGF-beta1) compared with WT cells and respond to infected erythrocytes or rIFN-gamma by releasing little NO. Moreover, IL-12p40KO macrophages had a severely impaired ability to internalize opsonized infected erythrocytes, suggesting that the low effector profile assumed by these cells may compromise antibody-mediated immunity. Taken together, our results support the idea that the absence of IL-12p40 not only affects IFN-gamma production but also has deep consequences in macrophage effector functions that may contribute to exacerbation of the early phase of P. c. chabaudi malaria. PMID- 12581494 TI - Effects of IFN-gamma and interleukin-1beta on major histocompatibility complex antigen and intercellular adhesion molecule-1 expression by 9L gliosarcoma: relevance to its cytolysis by alloreactive cytotoxic T lymphocytes. AB - To enhance the efficacy of cellular immunotherapy for gliomas, we tested the concept of using proinflammatory cytokine treatment with interferon-gamma (IFN gamma) or interleukin-1beta (IL-1beta) or both to render glioma cells more susceptible to cytolysis by alloreactive cytotoxic T lymphocytes (aCTL). The cytokines, separately or in combination, were able to upregulate major histocompatibility complex (MHC) class I antigen or intercellular adhesion molecule-1 (ICAM-1) on Fischer rat 9L gliosarcoma cells. 9L cells were incubated in vitro for 24, 48, or 72 h with varying concentrations of rat IFN-gamma (0-2000 U/ml) or recombinant human IL-1 (rHUIL-1) (0-1000 U/ml) or both. By 48 h, IFN gamma (500 U/ml) maximally induced the percentage of positive expressing cells and the relative antigen density of MHC class I and ICAM-1 on 9L cells, whereas IL-1 induced only ICAM-1 expression. Simultaneous incubation of IL-1 with IFN gamma did not further affect the induction of class I on 9L cells more than that achieved with IFN-gamma alone. 9L cells with upregulated MHC class I and ICAM-1 expression were more sensitive to lysis by aCTL in in vitro cytotoxicity assays, regardless of whether the precursor aCTL came from naive or from 9L-immunized rats. Furthermore, inhibition of 9L cytotoxicity in assays that included blocking antibodies to MHC class I or to ICAM-1 revealed that T cell receptor (TCR) interactions with MHC class I and that ICAM-1 interactions with lymphocyte function-associated-1 (LFA-1) antigen account for a portion of the glioma lysis by aCTL. PMID- 12581495 TI - Mechanisms of immunotherapeutic intervention by anti-CD154 (CD40L) antibody in high-risk corneal transplantation. AB - This study aimed to determine the effects of anti-CD154 on T cell cytokine profiles and ocular chemokine gene expression after high-risk corneal transplantation and to specifically determine if CD154 blockade is associated with a switch from a Th1 to a Th2 alloimmune response. Mice were used as recipients of syngeneic or multiple minor H or MHC antigen-mismatched corneal grafts. Recipient beds were neovascularized (high-risk). Hosts were randomized to receive either anti-CD154 antibody or control immunoglobulin (Ig) perioperatively. Two weeks after corneal transplantation, allospecific delayed type hypersensitivity (DTH) was evaluated. Frequencies of interferon-gamma (IFN gamma)-, interleukin-2 (IL-2)-, IL-4-, and IL-5-secreting T cells in the hosts were measured by enzyme-linked immunospot (ELISPOT) assay. Ocular chemokine gene expression in anti-CD154-treated and control hamster Ig-treated groups was determined using a multiprobe ribonuclease protection assay (RPA). Leukocyte infiltration of corneal grafts was evaluated microscopically. Anti-CD154-treated mice did not exhibit allospecific DTH. The frequencies of Th1 cytokine-producing but not Th2 cytokine-producing T cells were significantly reduced in anti-CD154 treated hosts. Postoperative mRNA levels of RANTES and macrophage inflammatory protein-1beta (MIP-1beta) in anti-CD154-treated eyes were substantially suppressed compared with hamster Ig-treated controls. Leukocyte infiltration was profoundly suppressed in grafts of anti-CD154-treated hosts. These data demonstrate that blockade of the CD40-CD154 costimulatory pathway after corneal transplantation inhibits Th1-mediated responses but does not induce a switch to a Th2-specific response. In addition, anti-CD154 therapy suppresses ocular chemokine gene expression and leukocytic infiltration into allografts. PMID- 12581496 TI - IFN-gamma induction by SCG, 1,3-beta-D-glucan from Sparassis crispa, in DBA/2 mice in vitro. AB - Sparassis crispa Fr. in an edible mushroom recently cultivable in Japan. A branched beta-glucan from S. crispa (SCG) is a major 6-branched 1,3-beta-D-glucan showing antitumor activity. In this study, we examined interferon-gamma (IFN gamma) induction by SCG from splenocytes in DBA/2 mice in vitro. In the splenocytes derived from almost all inbred strains of mice except for DBA/1 and DBA/2 mice, IFN-gamma production was not induced by SCG. The breeder and genders of DBA/2 mice showed no influence on IFN-gamma induction by SCG. On the other hand, the magnitude of IFN-gamma induction was lower in young mice than in their older counterparts. IFN-gamma was induced by SCG in adherent splenocytes, but IFN gamma production was most significantly increased by SCG in instances involving coexistence of adherent and nonadherent splenocytes. In fact, inhibition of cell cell contact reduced IFN-gamma induction by SCG. In addition, interleukin-12 p70 (IL-12p70) was induced by SCG in DBA/2 mice. It was suggested that soluble factors and cell-cell contact mediate synergistic effects on SCG-induced IFN gamma production. PMID- 12581497 TI - H(1)-Receptor activation triggers the endogenous nitric oxide signalling system in the rat submandibular gland. AB - BACKGROUND: Histamine is released from mast cells by immunologic and non immunologic stimuli during salivary gland inflammation, regulating salivary secretion. The receptor-secretory mechanism has not been studied in detail. AIMS: The studies reported were directed toward elucidating signal transduction/second messenger pathways within the rat submandibular gland associated with 2 thiazolylethylamine (ThEA)-induced H(1)-receptor responses. MATERIALS AND METHODS: To assess the H(1) receptor subtype expression in the rat submandibular gland, a radioligand binding assay was performed. The study also included inositolphosphates and cyclic GMP accumulation, protein kinase C and nitric oxide synthase activities, and amylase release. RESULTS: The histamine H(1) receptor subtype is expressed on the rat submandibular gland with high-affinity binding sites. The ThEA effect was associated with activation of phosphoinositide specific phospholipase C, translocation of protein kinase C, stimulation of nitric oxide synthase activity and increased production of cyclic GMP. ThEA stimulation of nitric oxide synthase and cyclic GMP was blunted by agents able to interfere with calcium movilization, while a protein kinase C inhibitor was able to stimulate ThEA action. On the other hand, ThEA stimulation evoked amylase release via the H1 receptor but was not followed by the L-arginine/nitric oxide pathway activation. CONCLUSIONS: These results suggest that, apart from the effect of ThEA on amylase release, it also appears to be a vasoactive chemical mediator that triggers vasodilatation, modulating the course of inflammation. PMID- 12581498 TI - Tumour necrosis factor-alpha, interleukin-2 soluble receptor and different inflammatory parameters in patients with rheumatoid arthritis. AB - BACKGROUND AND AIMS: Although the participation of cytokines in the pathogenesis of rheumatoid arthritis (RA) seems to be unequivocal, their relationship with current serum markers of this disease is not clear. The present study analyses whether there is any correlation between the levels of tumour necrosis factor alpha (TNF-alpha), interleukin-2 soluble receptor (sIL-2R) and the concentrations of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and beta(2) microglobulin in a group of 21 patients with RA, all rheumatoid factor positive. METHODS: The levels of TNF-alpha and sIL-2R were analysed in association with other parameters of inflammation (ESR, CRP and beta(2)-microglobulin). RESULTS: In comparison with the control group, RA patients presented high median levels of both cytokines, TNF-alpha (6.4 pg/ml) and sIL-2R (56 pmol/L), as well as of ESR (34 mm/h), CRP (0.9 mg/dl) and beta(2)-microglobulin (1.6 mg/dl) (p < 0.01). However, only ESR levels in the RA group significantly differ from the control group (p < 0.01). No correlation was found between the inflammatory parameters. CONCLUSIONS: These results suggested that TNF-alpha and slL-2R levels are up regulated in RA patients but did not significantly differ from the control group. Due to the chronic course of this disease, other inflammatory markers must be identified in order to provide early therapeutic strategies to these patients. PMID- 12581499 TI - Cell adhesion molecules involved in the leukocyte recruitment induced by venom of the snake Bothrops jararaca. AB - It has been shown that Bothrops jararaca venom (BjV) induces a significant leukocyte accumulation, mainly neutrophils, at the local of tissue damage. Therefore, the role of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1), LECAM-1, CD18, leukocyte function-associated antigen-1 (LFA-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1) on the BjV-induced neutrophil accumulation and the correlation with release of LTB4, TXA2, tumor necrosis factor-alpha, interleukin (IL)-1 and IL-6 have been investigated. Anti mouse LECAM-1, LFA-1, ICAM-1 and PECAM-1 monoclonal antibody injection resulted in a reduction of 42%, 80%, 66% and 67%, respectively, of neutrophil accumulation induced by BjV (250 microg/kg, intraperitoneal) injection in male mice compared with isotype-matched control injected animals. The anti-mouse CD18 monoclonal antibody had no significant effect on venom-induced neutrophil accumulation. Concentrations of LTB(4), TXA(2), IL-6 and TNF-alpha were significant increased in the peritoneal exudates of animals injected with venom, whereas no increment in IL-1 was detected. This results suggest that ICAM-1, LECAM-1, LFA-1 and PECAM 1, but not CD18, adhesion molecules are involved in the recruitment of neutrophils into the inflammatory site induced by BjV. This is the first in vivo evidence that snake venom is able to up-regulate the expression of adhesion molecules by both leukocytes and endothelial cells. This venom effect may be indirect, probably through the release of the inflammatory mediators evidenced in the present study. PMID- 12581500 TI - Gene expression of cell surface antigens in the early phase of murine influenza pneumonia determined by a cDNA expression array technique. AB - BACKGROUND: Influenza virus is a worldwide health problem with significant economic consequences. To study the gene expression pattern induced by influenza virus infection, it is useful to reveal the pathogenesis of influenza virus infection; but this has not been well examined, especially in vivo study. AIMS: To assess the influence of influenza virus infection on gene expression in mice, mRNA levels in the lung and tracheal tissue 48 h after infection were investigated by cDNA array analysis. METHODS: Four-week-old outbred, specific pathogen free strain, ICR female mice were infected by intra-nasal inoculation of a virus solution under ether anesthesia. The mice were sacrificed 48 h after infection and the tracheas and lungs were removed. To determine gene expression, the membrane-based microtechnique with an Atlas cDNA expression array (mouse 1.2 array II) was performed in accordance with the manual provided. RESULTS AND CONCLUSIONS: We focused on the expression of 46 mRNAs for cell surface antigens. Of these 46 mRNAs that we examined, four (CD1d2 antigen, CD39 antigen-like 1, CD39 antigen-like 3, CD68 antigen) were up-regulated and one (CD36 antigen) was down-regulated. Although further studies are required, these data suggest that these molecules play an important role in influenza virus infection, especially the phase before specific immunity. PMID- 12581501 TI - Pro-inflammatory cytokines in Turkish children with protein-energy malnutrition. AB - BACKGROUND: Protein-energy malnutrition (PEM) results from food insufficiency as well as from poor social and economic conditions. Development of PEM is due to insufficient nutrition. Children with PEM lose their resistance to infections because of a disordered immune system. It has been reported that the changes occurring in mediators referred to as cytokines in the immune system may be indicators of the disorders associated with PEM. AIMS: To determine the concentrations of pro-inflammatory cytokines in children with PEM, and to find out whether there was an association with the clinical presentation of PEM. METHODS: The levels of serum total protein, albumin, tumour necrosis factor alpha, and interleukin-6 were measured in 25 patients with PEM and in 18 healthy children as a control group. PEM was divided into two groups as kwashiorkor and marasmus. The kwashiorkor group consisted of 15 children and the marasmus group consisted of 10 children. RESULTS: Levels of serum total protein and albumin of the kwashiorkor group were significantly lower than both the marasmus group and controls (p < 0.05). In view of tumour necrosis factor-alpha levels, there was no difference between groups (p > 0.05). While levels of interleukin-6 in both the marasmus group and the kwashiorkor group were significantly higher compared with controls (p < 0.05), there was no significant difference between the groups of marasmus and kwashiorkor (p > 0.05). CONCLUSIONS: It was observed that the inflammatory response had increased in children with malnutrition. PMID- 12581502 TI - Control of particle size by coagulation of novel condensation aerosols in reservoir chambers. AB - The coagulation growth behavior of capillary aerosol generator (CAG) condensation aerosols was investigated in a series of reservoir chambers. Aerosols consisted of a condensed system of 0.7% w/w benzil (model drug) in propylene glycol (vehicle). These were generated into 250-, 500-, 1,000-, and 2,000-mL reservoirs in both flowing air-stream and static air experiments. Changes in drug and total aerosol particle size were measured by a MOUDI cascade impactor. In both series of experiments the CAG aerosols grew in size. Growth in flowing air-stream experiments was attributed to the amount of accumulation aerosols experienced in reservoirs during sampling and increased with increasing reservoir volume. Mean (SD) MMAD's for the total mass distribution measured for the 250- and 2,000-mL reservoirs were 0.70 (0.02) and 0.87 (0.03) microm, respectively. For the benzil mass distribution, they were 0.64 (0.02) and 0.87 (0.06) microm, respectively. Growth in static air experiments was dependent on the volume aerosol boluses were restricted to and increased with decreasing reservoir volume. Mean (SD) initial MMAD's for the benzil mass distribution for the 250- and 2,000-mL reservoirs were 1.44 (0.03) and 1.24 (0.08) microm, respectively. Holding aerosols for up to 60 sec further increased their size. Mean (SD) MMAD's for benzil after holding for 60 sec in these reservoirs were 2.28 (0.04) and 1.67 (0.09) microm, respectively. The coagulation behavior and therefore particle size of CAG aerosols may be modified and controlled by reservoir chambers for drug targeting within the respiratory tract. PMID- 12581503 TI - Cascade impactor data and the lognormal distribution: nonlinear regression for a better fit. AB - At present, cascade impactors are the instruments of choice for measuring the particle size distribution of aerosol present in the complex discharge from pharmaceutical inhalers. The distribution of drug captured in the cascade impactor may be most usefully represented by the lognormal distribution. Only two parameters must be extracted from the analysis of cascade impactor data in order to describe the distribution. These two parameters are the mass median aerodynamic diameter (MMAD) and the geometric standard deviation (GSD). A cumulative version of the lognormal curve or more frequently, a linearized version of the cumulative curve called a "log probability plot," is used as a surrogate for the lognormal curve. The probability plot has great appeal since a lognormal distribution yields a straight line on log probability paper. One may easily determine the apparent MMAD and GSD from this linear plot. However, when one plots a lognormal curve, using the MMAD and GSD derived from a log probability plot, over a histogram constructed from cascade impactor data, an obvious mismatch is frequently seen. In order to derive parameters that more truly reflect the impactor data, a computer program, which uses nonlinear regression to derive an MMAD and GSD for the lognormal curve, has been written. It is presented here. PMID- 12581504 TI - In vitro effect of a holding chamber on the mouth-throat deposition of QVAR (hydrofluoroalkane-beclomethasone dipropionate). AB - Experimental work has been conducted on the effect of an add-on holding chamber (Aerochamber) on the characteristics of deposition in a mouth-throat model using 100-microg hydrofluoroalkane-beclomethazone dipropionate (QVAR) metered dose inhalers at inhalation flow rates of 28.3, 60, and 90 L/min. A filter or cascade impactor downstream of the mouth-throat collected aerosol not depositing. The results emphasize the important well documented effect of a valved holding chamber (VHC), in reducing drug deposition in the mouth-throat. This reduction is largest (24% of nominal dose) at the lowest flow rate tested, becoming insignificant at 60 L/min. Total amount of drug delivered distal to the mouth throat increases with flow rate both with and without a holding chamber, increasing from 42% to 69% of the nominal dose without a VHC as the inspiratory flow rate increases from 28.3 to 90 L/min. The effect of the holding chamber on post mouth-throat delivery was small, reaching significance only at the highest flow rate (90 L/min), where an increase by 8% of the nominal dose was observed. No significant effect on MMAD of beclomethasone-dipropionate occurred when the holding chamber was used. An argument based on the interaction between induced turbulence and particle inertia is used to shed light on the above observations. PMID- 12581505 TI - Respiratory tract deposition efficiencies: evaluation of effects from smoke released in the Cerro Grande forest fire. AB - Forest-fire smoke inhaled by humans can cause various health effects. This smoke contains toxic chemicals and naturally occurring radionuclides. In northern New Mexico, a large wildfire occurred in May 2000. Known as the Cerro Grande Fire, it devastated the town of Los Alamos and damaged Los Alamos National Laboratory (LANL). Residents were concerned about the possible dissemination of radionuclides from LANL via smoke from the fire. To evaluate potential health effects of inhaling radionuclides contained in the smoke from the Cerro Grande Fire, it was first necessary to evaluate how much smoke would deposit in the human respiratory tract. The purpose of this study was to evaluate respiratory tract deposition efficiencies of airborne forest-fire smoke for persons of different ages exposed while inside their homes. Potential non-radiological health effects of a forest fire are reviewed. The deposition efficiencies presented can be used to evaluate in-home smoke deposition in the respiratory tract and expected radionuclide intake related to forest fires. The impact of smoke exposure on firemen fighting a forest fire is quantitatively discussed and compared. They primarily inhaled forest-fire smoke while outdoors where the smoke concentration was much higher than inside. Radionuclides released at the LANL site via the Cerro Grande Fire were restricted to naturally occurring radionuclides from burning trees and vegetation. Radiation doses from inhaled airborne radionuclides to individuals inside and outside the Los Alamos area were likely very small. PMID- 12581506 TI - Local versus total systemic bioavailability of beclomethasone dipropionate CFC and HFA metered dose inhaler formulations. AB - For inhaled formulations, the balance between desired local effects and undesired systemic activity can be expressed by L/T, where L represents bioavailability of drug from the lungs and T represents total systemic bioavailability. L/T is most useful when comparing formulations of the same inhaled substance. A high L/T is desirable as this implies efficient drug delivery to the target site, and minimization of unwanted activity from non-targeted drug delivery. The objective of this publication is to compare L/T for CFC and HFA inhaler formulations of beclomethasone dipropionate (BDP). Predictions of the L/T comparison were tested with clinical trials. From five deposition and pharmacokinetic studies, L/T ratios for CFC-BDP and HFA-BDP were calculated as 0.21 and 0.92, respectively. These ratios predicted two differences for the therapeutic use of these products: (1) a smaller dose of HFA-BDP than CFC-BDP may be required for efficacy; and (2) a smaller number of adverse events may be observed for the HFA-BDP product, when delivered at the equivalent dose, compared to the CFC comparitor. A dose-response study confirmed that less than half the dose of HFA-BDP is needed to give the same efficacy as CFC-BDP. Two safety studies that measured adrenal suppression demonstrated less suppression with HFA-BDP than with a comparable efficacious dose of CFC-BDP. It is concluded that L/T is a useful parameter that incorporates the systemic contributions of lung deposition and pharmacokinetics. It is recommended that this parameter be considered whenever deposition and pharmacokinetic data for two formulations of the same inhaled substance are compared. PMID- 12581507 TI - Effects on lung function, symptoms, and bronchial hyperreactivity of low-dose inhaled beclomethasone dipropionate given with HFA-134a or CFC propellant. AB - The aim of this study was to compare the efficacy of BDP 200 microg bid via metered dose inhaler, using HFA-134a (Chiesi Farmaceutici S.p.A., Parma, Italy) versus CFC (Becotide, Allen & Hanburys, U.K.) as a propellant. 172 adult patients (86 in each group) with stable mild persistent asthma who completed a 7-day run in period were randomized to receive a 6-week treatment in a double-blind, double dummy, parallel-group design; 164 patients completed the study. Morning and evening PEFR, use of rescue salbutamol, number of day-time and night-time asthma attacks, number of night-time awakenings and clinical symptoms were recorded daily on a diary card. Pulmonary function tests (FEV(1), FVC, PEFR, and MEF(50)) were measured at the clinic before and after the 1-week run-in period, and after 3 and 6 weeks of treatment. A challenge test with inhaled methacholine was completed at baseline and at the end of the treatment period to assess potential bronchial hyper-reactivity in a subgroup of subjects (n = 65; 34 HFA, 31 CFC). In accordance with asthma of mild severity (FEV(1) predicted over 90% in both groups), a small improvement in lung function compared to baseline was seen for both treatments, significantly for FEV(1) in BDP HFA and MEF(50) in both groups. The two formulations of BDP had similar efficacy for the primary outcome variable morning PEFR (ITT population mean difference 5.8 L/min; C.I. -4.9 to +16.5) as well as for the secondary outcomes of evening PEFR and clinic FEV(1). There were small improvements in methacholine PD(20) and PC20 in both groups, with no significant difference between treatments. A total of 22 and 19 drug-related adverse events were reported in the BDP HFA and CFC groups, respectively; most events were of seasonal nature or were local effects due to the use of inhaled corticosteroids. It can be concluded that the newly developed formulation of BDP given via HFA-134a seems to provide similar asthma control, compared with the same low daily dose of the active drug delivered via CFC. Further studies are needed using higher doses in moderate to severe asthma to confirm these preliminary findings. PMID- 12581508 TI - Allergen challenge and deposition of nedocromil sodium in asthma. AB - We examined whether the acute protective effect of nedocromil sodium aerosol could be enhanced by increasing the deposition uniformity of the drug in the lungs of adult patients with allergic asthma. Ten patients with mild-to-moderate asthma were challenged with the same doses of allergen on two occasions in a randomized manner. Thirty minutes before these challenges, patients inhaled 4 mg nedocromil sodium, admixed with the radioisotope (99m)technetium. Radiolabeled drug was inhaled during slow (25.4 +/- 4.6 L/min) and faster (58.0 +/- 7.3 L/min) inhalations from a 700 ml holding chamber. Percent changes in FEV(1) at the same top dose of allergen on the two treatment visits were compared. Lung deposition fraction (LDF) and indices of distribution uniformity, quantified from gamma camera images, were also compared. Acute protection against allergen challenge was similar and complete after slow or faster inspiration of nedocromil sodium. Mean (+/- SD) allergen-induced changes in FEV(1) were -1.05 +/- 2.78% and -0.39 +/- 2.80%, respectively, compared to -26.30 +/- 8.49% on a screening challenge (no drug). Mean LDF was also similar on the two visits, averaging 16.4 +/- 4.6% and 16.1 +/- 7.2% of administered drug, respectively. Distribution of nedocromil sodium was most uniform after slow inspiration, but increased uniformity was not related to enhanced protection. Complete protection against acute bronchoconstriction induced by inhaled allergen can be obtained with 4 mg of nedocromil sodium aerosol, inhaled from a large volume holding chamber, 30 min before the exposure, and at inspiratory flow rates between approximately 20-60 L/min. Protection does not appear to be enhanced by increased uniformity of drug distribution within the lungs. PMID- 12581509 TI - Performance of large- and small-volume valved holding chambers with a new combination long-term bronchodilator/anti-inflammatory formulation delivered by pressurized metered dose inhaler. AB - The treatment of both the bronchoconstriction and inflammatory aspects of asthma simultaneously by a single pressurized metered dose inhaler (pMDI) represents a significant advance in convenience to the patient. However, a valved holding chamber (VHC) may still be needed to reduce the coarse component of the dose that is likely to deposit in the oropharyngeal region, and a small sized device may offer significant advantages to the patient from the standpoint of compliance with therapy. VHCs representing small (adult AeroChamber Plus with mouthpiece, 149-mL) and large (Volumatic, 750-mL) devices have been compared in an in vitro evaluation with Seretide/Advair (hydro-fluoro alkane [HFA]-formulated fluticasone propionate [FP = 125 microg/dose] and salmeterol xinafoate [SX = 25 microg/dose]) by Andersen Mark-II eight-stage impactor operated at 28.3 L/min following compendial methodology. Fine particle fraction, based on the size range from 1.1 to 4.7 microm aerodynamic diameter, from either large or small VHCs with either component (69-79%) was similar [p > or = 0.08], and significantly greater than that from the pMDI alone (approximately 40%) [p < 0.001]. Fine particle dose emitted by the VHCs for SX (8.2 +/- 0.8 microg for the AeroChamber Plus and 7.7 +/- 0.5 microg for the Volumatic) were comparable, and also similar to the fine particle dose delivered by the pMDI when used without a VHC (7.6 +/- 0.6 microg). Fine particle doses for the FP component delivered by the two VHCs (46.4 +/- 3.4 microg for the AeroChamber Plus and 46.3 +/- 2.7 microg for the Volumatic) were equivalent, but were slightly greater than the corresponding fine particle dose from the pMDI alone (39.1 +/- 2.6 microg). However, this difference (approximately 20%) is close to the limit of resolution based on intermeasurement variability and is unlikely to have clinical significance, given the interpatient variability seen with inhaled drug therapy. It is therefore concluded that either of these VHCs has equivalent in vitro performance with this combination formulation in terms of the portion of the dose emitted from the pMDI that is likely to reach the receptors in the lungs. PMID- 12581510 TI - In vitro aerosol performance and dose uniformity between the Foradile Aerolizer and the Oxis Turbuhaler. PMID- 12581511 TI - HIV type 1 infection alters cytokine mRNA expression in thymus. AB - We and others have previously shown that HIV infection of human thymus/liver implants in severe combined immunodeficient (SCID-hu Thy/Liv) mice results in a loss of CD4(+) thymocytes and a decreased recovery of human myeloid and erythroid colony-forming activity. Furthermore, our previous studies have shown that this decrease in colony-forming potential is due to indirect effects, rather than to massive loss of CD34(+) hematopoietic progenitor cells, suggesting that HIV infection might alter expression of hematopoietic cytokines. Herein we have investigated potential HIV-1-induced perturbations of hematolymphoid microenvironments by determining the effect of HIV-1 infection on levels of cytokine mRNAs in human stroma and thymocytes, using the reverse transcription polymerase chain reaction (RT-PCR). The levels of interleukin 6 (IL-6), interferon gamma (IFN-gamma), and IL-2 RNAs increased and macrophage inflammatory protein 1beta (MIP-1beta) RNA decreased significantly in infected thymocytes. IL 6 RNA levels in stroma also increased somewhat with infection; however, expression of stromal cell-derived factor 1 (SDF-1) by stromal elements was not affected. IL-4 RNA levels were unaffected by infection in both stroma and thymocytes. Antiretroviral drug treatment of the infected animals, which results in restoration of colony-forming potential, tends to restore the cytokine mRNA levels in HIV-1-infected implants to those of mock-infected implants. These results indicate that HIV-1 infection can greatly distort the cytokine profiles in Thy/Liv implants, and suggest that cytokine perturbation could be involved in virus-induced inhibition of hematopoiesis. PMID- 12581513 TI - HIV-mediated gammadelta T cell depletion is specific for Vgamma2+ cells expressing the Jgamma1.2 segment. AB - Circulating Vgamma2/Vdelta2(+) T cells, normally constituting 3-6% of all CD3(+) T cells in blood, are severely depleted after HIV infection. The mechanism(s) for Vgamma2/Vdelta2(+) T cell depletion are unknown, partly because these cells are CD4(-) and resistant to HIV infection. To determine whether this cell depletion was general for all Vgamma2(+) cells or specific for an individual subset, we analyzed the Vgamma2 repertoire and found consistent differences between HIV(+) and uninfected control samples. The change in Vgamma2 repertoire was the result of preferentially depleting only those Vgamma2 cells that express the Jgamma1.2 segment. The specific loss of Vgamma2-Jgamma1.2(+) cells was polyclonal, as the Vgamma subset retained normal diversity even after HIV infection, and loss occurred without significant changes in the paired chain (Vdelta2) repertoire, or in the alternate Vdelta1 chain repertoire. Specific depletion of Vgamma2 Jgamma1.2/Vdelta2 T cells is the first evidence of a common, T cell receptor dependent cell loss in HIV disease and it provides a clear example of bystander cell depletion. Vgamma2-Jgamma1.2/Vdelta2 T cells mediate potent responses to microbial pathogens including HIV, and loss of this subset is an important aspect of AIDS pathogenesis. PMID- 12581512 TI - Follow-up of antiretroviral treatment in liver transplant recipients with primary and chronic HIV type 1 infection. AB - The prognosis of HIV-1-infected patients has dramatically improved but progression to liver failure occurs now frequently in subjects coinfected with hepatitis C virus (HCV). This has raised the issue of organ transplantation, but the knowledge about the effect of concomitant antiretroviral and immunosuppressive therapy is limited. The objective of the study was to describe viral and immunological events in antiretroviral-treated orthotopic liver transplant (OLT) recipients with primary (PHI) or chronic HIV-1 infection. Three HIV-1-infected patients with liver cirrhosis due to chronic HCV infection underwent OLT. A fourth patient developed PHI at OLT. Immunosuppressive drugs and combination antiretroviral therapy were given. The effects on HIV-1 load, viral diversity and divergence, and CD4(+) T cell counts, were studied. One patient died after 3 months. Three subjects were alive after 9 months, 14 months, and 3 years, respectively. In the PHI patient, viral load decreased during the second week of illness despite immunosuppression. During the third week the viremia increased until antiretroviral treatment was initiated. In all four patients, the HIV-1 replication was effectively inhibited during follow-up by the treatment, as determined by undetectable plasma viremia, lack of viral sequence changes, and increase in CD4(+) T cells. The pattern of viral dynamics may suggest that the innate immunity causes the earliest decline of viral load in PHI patients. A lack of adaptive immunity may thereafter lead to an increase in viremia in heavily immunosuppressed individuals. However, a specific HIV-1 immunity is not necessary to efficiently inhibit the viral replication when potent antiretroviral therapy is given in liver transplant recipients with primary or chronic HIV-1 infection. PMID- 12581514 TI - Expression patterns of phenotypic markers on lymphocytes from human immunodeficiency virus type 2-infected baboons. AB - The development of AIDS in HIV-1-infected humans is associated with profound changes in the expression patterns of lymphocyte phenotypic markers associated with increased immune activation and with decreased recall immune responses. In assessing these immunologic changes in an animal model, we characterized the expression patterns of immune activation markers on lymphocyte subsets during the acute, chronic, and end stages of HIV-2 infection in baboons. Using flow cytometry, we identified 21 human-specific monoclonal antibodies that were cross reactive with baboon lymphocytes; however, expression of only 2 of these markers was altered significantly after HIV-2 infection. We found an increase in baboon class II antigen (as measured by anti-HLA-DR) in the CD4(+) T cell subset within 8 weeks of infection (p = 0.045). Moreover, after 1 year of infection, CD11b was downregulated on CD8(+) T lymphocytes (p = 0.027). This downregulation of CD11b was consistently observed in all of the groups of baboons that were chronically infected with three different HIV-2 isolates. In addition, we found substantial downregulation of the interleukin 2 receptor (CD25) and upregulation of class II antigen on CD8(+) lymphocytes in a baboon with an AIDS-like disease. These and other phenotypic markers of immune activation may facilitate characterization of the immunopathogenesis of AIDS in nonhuman primate animal models. PMID- 12581517 TI - Circadian clocks: a tale of two feedback loops. AB - Circadian clocks in a wide range of organisms are thought to consist of two interdependent transcriptional feedback loops. In Drosophila, the first loop has been well characterized and controls rhythmic period expression. In this issue of Cell, define a role for a transcriptional activator and a repressor in the second feedback loop. PMID- 12581516 TI - Bitter-sweet solution in taste transduction. AB - A contentious issue in taste research might have come to a close. Zhang et al., in this issue of Cell, provide broad support for the notion that the recognition of sweet, umami, and bitter tastes use the same signaling molecules. Moreover, they show that individual taste cells are dedicated to the transduction of only one of these three taste qualities. PMID- 12581518 TI - Better chemistry for better survival, through regulation. AB - In this issue of Cell, report on new aspects of the regulation of yeast ribonucleotide reductase, the mechanism by which dNTP levels are increased following DNA damage, and the consequences of the metabolic changes. PMID- 12581519 TI - Coaxing the LDL receptor family into the fold. AB - Low-density-lipoprotein (LDL) receptor family members control diverse developmental and physiological pathways. In this issue of Cell, both Culi and Mann and Hsieh et al. report on Boca/MESD, a highly conserved chaperone required for transport of LDLR family proteins to the cell surface. Together with recent insights into the atomic structure of the LDL receptor, they shed new light on the synthesis and trafficking of this important class of multifunctional receptors. PMID- 12581520 TI - Coding of sweet, bitter, and umami tastes: different receptor cells sharing similar signaling pathways. AB - Mammals can taste a wide repertoire of chemosensory stimuli. Two unrelated families of receptors (T1Rs and T2Rs) mediate responses to sweet, amino acids, and bitter compounds. Here, we demonstrate that knockouts of TRPM5, a taste TRP ion channel, or PLCbeta2, a phospholipase C selectively expressed in taste tissue, abolish sweet, amino acid, and bitter taste reception, but do not impact sour or salty tastes. Therefore, despite relying on different receptors, sweet, amino acid, and bitter transduction converge on common signaling molecules. Using PLCbeta2 taste-blind animals, we then examined a fundamental question in taste perception: how taste modalities are encoded at the cellular level. Mice engineered to rescue PLCbeta2 function exclusively in bitter-receptor expressing cells respond normally to bitter tastants but do not taste sweet or amino acid stimuli. Thus, bitter is encoded independently of sweet and amino acids, and taste receptor cells are not broadly tuned across these modalities. PMID- 12581521 TI - TGF-beta signaling activates steroid hormone receptor expression during neuronal remodeling in the Drosophila brain. AB - Metamorphosis of the Drosophila brain involves pruning of many larval-specific dendrites and axons followed by outgrowth of adult-specific processes. From a genetic mosaic screen, we recovered two independent mutations that block neuronal remodeling in the mushroom bodies (MBs). These phenotypically indistinguishable mutations affect Baboon function, a Drosophila TGF-beta/activin type I receptor, and dSmad2, its downstream transcriptional effector. We also show that Punt and Wit, two type II receptors, act redundantly in this process. In addition, knocking out dActivin around the mid-third instar stage interferes with remodeling. Binding of the insect steroid hormone ecdysone to distinct ecdysone receptor isoforms induces different metamorphic responses in various larval tissues. Interestingly, expression of the ecdysone receptor B1 isoform (EcR-B1) is reduced in activin pathway mutants, and restoring EcR-B1 expression significantly rescues remodeling defects. We conclude that the Drosophila Activin signaling pathway mediates neuronal remodeling in part by regulating EcR-B1 expression. PMID- 12581522 TI - The fragile X syndrome protein FMRP associates with BC1 RNA and regulates the translation of specific mRNAs at synapses. AB - The Fragile X syndrome, which results from the absence of functional FMRP protein, is the most common heritable form of mental retardation. Here, we show that FMRP acts as a translational repressor of specific mRNAs at synapses. Interestingly, FMRP associates not only with these target mRNAs, but also with the dendritic, non-translatable RNA BC1. Blocking of BC1 inhibits the interaction of FMRP with its target mRNAs. Furthermore, BC1 binds directly to FMRP and can also associate, in the absence of any protein, with the mRNAs regulated by FMRP. This suggests a mechanism where BC1 could determine the specificity of FMRP function by linking the regulated mRNAs and FMRP. Thus, when FMRP is not present, loss of translational repression of specific mRNAs at synapses could result in synaptic dysfunction phenotype of Fragile X patients. PMID- 12581523 TI - vrille, Pdp1, and dClock form a second feedback loop in the Drosophila circadian clock. AB - The Drosophila circadian clock consists of two interlocked transcriptional feedback loops. In one loop, dCLOCK/CYCLE activates period expression, and PERIOD protein then inhibits dCLOCK/CYCLE activity. dClock is also rhythmically transcribed, but its regulators are unknown. vrille (vri) and Par Domain Protein 1 (Pdp1) encode related transcription factors whose expression is directly activated by dCLOCK/CYCLE. We show here that VRI and PDP1 proteins feed back and directly regulate dClock expression. Repression of dClock by VRI is separated from activation by PDP1 since VRI levels peak 3-6 hours before PDP1. Rhythmic vri transcription is required for molecular rhythms, and here we show that the clock stops in a Pdp1 null mutant, identifying Pdp1 as an essential clock gene. Thus, VRI and PDP1, together with dClock itself, comprise a second feedback loop in the Drosophila clock that gives rhythmic expression of dClock, and probably of other genes, to generate accurate circadian rhythms. PMID- 12581524 TI - Boca, an endoplasmic reticulum protein required for wingless signaling and trafficking of LDL receptor family members in Drosophila. AB - The maturation of cell surface receptors through the secretory pathway often requires chaperones that aid in protein folding and trafficking from one organelle to another. Here we describe boca, an evolutionarily conserved gene in Drosophila melanogaster, which encodes an endoplasmic reticulum protein that is specifically required for the intracellular trafficking of members of the low density lipoprotein family of receptors (LDLRs). Two LDLRs in flies, Arrow, which is required for Wingless signal transduction, and Yolkless, which is required for yolk protein uptake during oogenesis, both require boca function. Consequently, boca is an essential component of the Wingless pathway but is more generally required for the activities of multiple LDL receptor family members. PMID- 12581525 TI - Mesd encodes an LRP5/6 chaperone essential for specification of mouse embryonic polarity. AB - Specification of embryonic polarity and pattern formation in multicellular organisms requires inductive signals from neighboring cells. One approach toward understanding these interactions is to study mutations that disrupt development. Here, we demonstrate that mesd, a gene identified in the mesoderm development (mesd) deletion interval on mouse chromosome 7, is essential for specification of embryonic polarity and mesoderm induction. MESD functions in the endoplasmic reticulum as a specific chaperone for LRP5 and LRP6, which in conjunction with Frizzled, are coreceptors for canonical WNT signal transduction. Disruption of embryonic polarity and mesoderm differentiation in mesd-deficient embryos likely results from a primary defect in WNT signaling. However, phenotypic differences between mesd-deficient and wnt3(-)(/)(-) embryos suggest that MESD may function on related members of the low-density lipoprotein receptor (LDLR) family, whose members mediate diverse cellular processes ranging from cargo transport to signaling. PMID- 12581526 TI - Initiation of RPS2-specified disease resistance in Arabidopsis is coupled to the AvrRpt2-directed elimination of RIN4. AB - Plants have evolved a sophisticated innate immune system to recognize invading pathogens and to induce a set of host defense mechanisms resulting in disease resistance. Pathogen recognition is often mediated by plant disease resistance (R) proteins that respond specifically to one or a few pathogen-derived molecules. This specificity has led to suggestions of a receptor-ligand mode of R protein function. Delivery of the bacterial effector protein AvrRpt2 by Pseudomonas syringae specifically induces disease resistance in Arabidopsis plants expressing the RPS2 R protein. We demonstrate that RPS2 physically interacts with Arabidopsis RIN4 and that AvrRpt2 causes the elimination of RIN4 during activation of the RPS2 pathway. AvrRpt2-mediated RIN4 elimination also occurs in the rps2, ndr1, and Atrar1 mutant backgrounds, demonstrating that this activity can be achieved independent of an RPS2-mediated signaling pathway. Therefore, we suggest that RPS2 initiates signaling based upon perception of RIN4 disappearance rather than direct recognition of AvrRpt2. PMID- 12581527 TI - Arabidopsis RIN4 is a target of the type III virulence effector AvrRpt2 and modulates RPS2-mediated resistance. AB - Type III pili deliver effector proteins (virulence factors) from bacterial pathogens to host cells. Plants express disease resistance (R) proteins that respond specifically to a particular type III effector by activating immune responses. We demonstrated previously that two unrelated type III effectors from Pseudomonas syringae target and modify the Arabidopsis RIN4 protein. Here, we show that AvrRpt2, a third, unrelated type III effector, also targets RIN4 and induces its posttranscriptional disappearance. This effect is independent of the presence of RPS2, the Arabidopsis R protein that senses AvrRpt2. RIN4 overexpression inhibits multiple phenotypes associated with AvrRpt2 function. Conversely, disruption of RIN4 results in RPS2-dependent lethality. RPS2 and RIN4 physically associate in the plant. We suggest that RIN4 is the target of the AvrRpt2 virulence function, and that perturbation of RIN4 activates RPS2. Thus, RIN4 is a point of convergence for the activity of at least three unrelated P. syringae type III effectors. PMID- 12581528 TI - Survival of DNA damage in yeast directly depends on increased dNTP levels allowed by relaxed feedback inhibition of ribonucleotide reductase. AB - In eukaryotes, DNA damage elicits a multifaceted response that includes cell cycle arrest, transcriptional activation of DNA repair genes, and, in multicellular organisms, apoptosis. We demonstrate that in Saccharomyces cerevisiae, DNA damage leads to a 6- to 8-fold increase in dNTP levels. This increase is conferred by an unusual, relaxed dATP feedback inhibition of ribonucleotide reductase (RNR). Complete elimination of dATP feedback inhibition by mutation of the allosteric activity site in RNR results in 1.6-2 times higher dNTP pools under normal growth conditions, and the pools increase an additional 11- to 17-fold during DNA damage. The increase in dNTP pools dramatically improves survival following DNA damage, but at the same time leads to higher mutation rates. We propose that increased survival and mutation rates result from more efficient translesion DNA synthesis at elevated dNTP concentrations. PMID- 12581529 TI - Nitroxide radicals protect against DNA damage in rat epithelial cells induced by nitric oxide, nitroxyl anion and peroxynitrite. AB - In order to gain more knowledge on the antioxidant role of nitroxide radicals, in this study we investigate their possible protective action against DNA damage induced by nitric oxide (NO) and reactive nitrogen oxide species deriving from it, namely nitroxyl anion (NO(-)) and peroxynitrite (ONOO(-)). Rat trachea epithelial cells were exposed under aerobic conditions to (1) NO generated by 150 microM S-nitrosoglutathione monoethyl ester (GSNO-MEE), (2) NO(-) generated by 200 microM Angeli's salt (Na(2)N(2)O(3)) (3) ONOO(-) generated by 1mM SIN-1 (3 morpholino-sydnonimine) and (4) 100 microM synthesized ONOO(-), in the absence and presence of 5 microM of two indolinonic nitroxides synthesized by us and the piperidine nitroxide TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl). DNA damage was assessed using the comet assay-a rapid and sensitive, single-cell gel electrophoresis technique used to detect primary DNA damage in individual cells. The parameter tail moment, used as an index of DNA damage, showed that in all cases the nitroxides remarkably inhibited DNA strand breaks induced by the different nitrogen oxide species. All three nitroxides protect to the same extent, except in the case of synthesized peroxynitrite where the aromatic nitroxides 1 and 2 are more efficient than TEMPO. These findings are consistent with the antioxidant character of nitroxide compounds and give additional information on the potential implications for their use as therapeutic agents. PMID- 12581530 TI - In vitro genotoxicity of lead acetate: induction of single and double DNA strand breaks and DNA-protein cross-links. AB - Lead is present in the natural and occupational environment and is reported to interact with DNA, but the mechanism of this interaction is not fully understood. Using the alkaline comet assay we showed that lead acetate at 1-100 microM induced DNA damage in isolated human lymphocytes measured the change in the comet tail length. At 1 and 10 microM we observed an increase in the tail length, whereas at 100 microM a decrease was seen. The former effect could follow from the induction of DNA strand breaks and/or alkali-labile sites (ALS), the latter from the formation of DNA-DNA and/or DNA-protein cross-links. No difference was observed between tail length for the alkaline and pH 12.1 versions of the assay, which indicates that strand breaks and not ALS are responsible for the tail length increase induced by lead. The neutral version of the test revealed that lead acetate induced DNA double-strand breaks at all concentrations tested. The presence of spin traps, 5,5-dimethyl-pyrroline N-oxide (DMPO) and N-tert-butyl alpha-phenylnitrone (PBN) did not influence the level of DNA damage induced by lead. Post-treatment of the lead-damaged DNA (at 100 microM treatment concentration) by endonuclease III (Endo III) and formamidopyrimidine-DNA glycosylase (Fpg), enzymes recognizing oxidized DNA bases, as well as 3 methyladenine-DNA glycosylase II, an enzyme recognizing alkylated bases, gave rise to a significant increase in the extent of DNA damage. Proteinase K caused an increase in comet tail length, suggesting that lead acetate might cross-link DNA with nuclear proteins. Vitamin A, E, C, calcium chloride and zinc chloride acted synergistically on DNA damage evoked by lead. The results obtained suggest that lead acetate may induce single-strand breaks (SSB) and double-strand breaks (DSB) in DNA as well as DNA-protein cross-links. The participation of free radicals in DNA-damaging potential of lead is not important and it concerns other reactive species than could be trapped by DMPO or PBN. PMID- 12581531 TI - Spontaneous mutations in the Helicobacter pylori rpsL gene. AB - Several studies have revealed that the Helicobacter pylori genome differs markedly from strain to strain, perhaps as a result of mutations arising during persistent infection and/or related to the observed variation in virulence. The development of a detection system for mutations in H. pylori genes might therefore help us to develop a better understanding of its mutability, and in this way help us to develop plans for investigating the relationship between its genomic variability and the pathogenesis of various gastric and duodenal diseases associated with the long-term H. pylori infections. We have therefore begun a study of H. pylori mutability using the endogenous rpsL gene as a marker. Spontaneous mutant frequencies were measured and compared among H. pylori strains, after incubation on plates containing 50 microg/ml of streptomycin for 10 days as a selection procedure. The rpsL gene of each streptomycin-resistant (Str(r)) mutant was amplified by polymerase-chain-reaction (PCR) and sequenced. All of the mutations we characterized were localized at codons 43 or 88 of the rpsL gene and were base transitions from A to G, replacing lysine with arginine. This is in contrast to the spontaneous Str(r) mutants isolated from Escherichia coli, which resulted from either A to G transitions at lysine codons 42 and 87, or A to T or C transversions at lysine codon 42. The spontaneous mutant frequencies of the rpsL gene in H. pylori were of the order of 10(-9), and there were significant differences in spontaneous mutant frequencies among the strains tested. This mutation detection system might be of value in screening clinical isolates for H. pylori mutator phenotypes. PMID- 12581532 TI - Cytogenetic effects in workers occupationally exposed to tobacco dust. AB - Tobacco dust mainly contains nitrosamines, which are readily absorbed by the body tissues like skin, respiratory epithelium, and mucous membrane of mouth, nose and intestines. Exposure to tobacco dust is known to affect the respiratory tracts in humans. In the present study, cytogenetic effects of exposure to tobacco dust are evaluated in 154 male tobacco factory workers and 138 age and sex matched controls by analysing chromosomal aberrations in their peripheral blood lymphocytes. The workers were in the age group of 20-55 years and were employed in the tobacco processing factory for 1-32 years. Heparinised blood samples were collected from workers and control subjects and lymphocyte cultures were carried out by using standard technique. Slides were prepared and 150 metaphases were screened for each sample for various structural and numerical types of abnormalities. A statistically significant increase was observed in the frequencies of chromosomal aberrations in non-smoking and smoking exposed groups when compared to the respective controls. An increase in the frequencies of chromosomal aberrations was also observed with increase in years of service in the exposed subjects. PMID- 12581533 TI - Polycyclic aromatic hydrocarbon-DNA adducts in human sperm as a marker of DNA damage and infertility. AB - Severe DNA damage, which might prevent egg fertilization or the development of the embryo, could be a cause of infertility. In order to assess whether polycyclic aromatic hydrocarbon (PAH)-DNA adducts are an early marker of sperm genotoxicity and infertility, we studied 205 men consecutively recruited from 1 January to 30 May 2001 through the Infertility Clinic of the University of Milan (Italy), with morphological abnormalities in the sperm. No known causes of infertility were present in their female partners. Sperm were collected after 3-5 days of abstinence, fixed on polylysine slides, and frozen at -20 degrees C. PAH DNA adducts were measured by immunofluorescence using a polyclonal antiserum. A questionnaire was filled out at the time of the visit, with demographic information, smoking and drinking habits, and occupational history. Data on PAH DNA adducts were available for 182 men. The mean age of the subjects was 35.5+/ 5.0 years; 38.6% of them were current smokers. PAH-DNA adducts were negatively correlated with the percentage of physiologic forms (r=-0.18; P=0.016) and with abnormalities of the neck of the sperm cell (r=-0.21; P>/=0.009), while they were positively correlated with morphological abnormalities of the head (r=0.30; P>0.0001). Occupational exposure to PAH, but not smoking, was significantly associated with higher levels of PAH-DNA adducts. A significant negative association was observed between daily alcohol consumption and PAH-DNA adducts in sperm (P=0.01). PAH-DNA adducts levels were significantly higher in infertile versus fertile men (P=0.04). These results suggest a role for DNA damage in infertility. PMID- 12581534 TI - Mutagenic and cytotoxic effectiveness of diisopropyl xanthogen polysulphide in human lymphocyte cultures. AB - The mutagenic and cytotoxic effectiveness of the new rubber vulcanisation accelerator diisopropyl xanthogen polysulphide (Robac AS 100) was tested in human lymphocyte cultures of four healthy probands. The concentrations of Robac AS 100 were 0.57, 5.7 and 57.0 microg/ml. Higher concentrations showed too high cytotoxicity to be evaluable. Without external activation, incubation time with Robac AS 100 was 21 h. In the presence of rat liver microsomes from aroclor induced rats (2mg microsomal protein/ml), incubation of the test compound was 2h. Mutagenicity testing was performed by analysis of micronuclei (MN), structural chromosome aberrations (CAs) and sister chromatid exchanges (SCEs). The MN-rate was determined using the cytochalasin B (cyt B) block method. For evaluation of cytotoxicity, mitotic index (MI) and nuclear division index (NDI) were determined. The validity of the test methods was ascertained by positive controls: mitomycin C (MMC) and bleomycin (BLM) were used in experiments without exogenous activation and cyclophosphamide (CP) in experiments with exogenous activation. The presence of rat liver microsomes increased the mutagenic effect of Robac AS 100 in the SCE- and MN-test. But only the highest Robac AS 100 concentration (57.0 microg/ml) showed significantly increased mutagenic activity in all tests. However, cytotoxicity at this concentration was already substantial. Therefore, we consider the evidence for mutagenicity of Robac AS 100 as limited. PMID- 12581535 TI - Modulation of mutagenicity of various mutagens by lignin derivatives. AB - The effect of lignin on cytotoxicity, mutagenicity and SOS response induced by 4 nitroquinoline-N-oxide (4NQO), 3-(5-nitro-2-furyl)acrylic acid (5NFAA), 2 nitrofluorene (2NF) as well as hydrogen peroxide was investigated in bacterial assay systems, i.e. the Ames test with Salmonella typhimurium TA98, TA100, TA102 and the SOS chromotest with Escherichia coli PQ37. Lignin preparations obtained from beech wood significantly decreased the mutagenicity induced by 4NQO, 2NF and H(2)O(2). In the case of mutagenicity induced by 5NFAA the effect was lower. Antimutagenic properties of lignin samples tested were shown also by SOS chromotest where lignin inhibited the ability of both 4NQO and H(2)O(2) to induce the SOS response. Derivatives of lignin including those from soft and hard wood, as well as from annual plants differ in their efficiency to inhibit the induction of the SOS response. The modified lignins isolated from beech and spruce wood exhibit a high level of protection. Lignins from annual plants-corn cobs and straw-only marginally evoked an antimutagenic response, but their effect was increased by hydrothermic treatment of both annual plants. The results obtained indicate the prospective utilization of lignin preparations as additive in chemo prevention. The antimutagenic effect of lignin samples varies with the method of isolation and modification, as well as with the genetic origin of the lignin. PMID- 12581536 TI - Association of hepatitis virus infection, alcohol consumption and plasma vitamin A levels with urinary 8-hydroxydeoxyguanosine in chemical workers. AB - Urinary 8-hydroxydeoxyguanosine (8-OHdG) DNA adduct has been used as a biomarker in epidemiological studies. However, the determinants for urinary 8-OHdG have not been clearly identified. We tested urinary 8-OHdG levels in 205 male workers who had been exposed to vinyl chloride monomer (VCM). Epidemiological information was obtained by an interviewer-administered questionnaire. Hepatitis B surface antigen (HBsAg) and anti-hepatitis C antibody (anti-HCV) were also determined by immunoassay. Plasma antioxidants including Vitamins A and E, alpha- and beta carotenes were assayed by high performance liquid chromatography. Median of urinary 8-OHdG level was 9.8 ng/mg creatinine (range, 1.4-60.1). Multiple linear regression analysis showed that alcohol drinkers had higher urinary 8-OHdG than those who did not, but there was no dose-response between the amount of alcohol consumption and urinary 8-OHdG. Workers with positive HBsAg, anti-HCV and elevated plasma Vitamin A level were independently associated with higher levels of urinary 8-OHdG, whereas age, smoking, body mass index, plasma alpha- and beta carotenes, Vitamin E levels, or VCM exposure did not show such an association. The results suggest that active inflammation of hepatitis B and C, alcohol consumption and higher Vitamin A level can induce oxidative stress. Thus, we conclude that potential determinants need to be considered in epidemiological studies when urinary 8-OHdG is used as a biomarker. PMID- 12581537 TI - DNA damage induced by indirect and direct acting mutagens in catalase-deficient transgenic tobacco. Cellular and acellular Comet assays. AB - We have measured the level of DNA damage induced by treating roots (cellular Comet assay) and isolated root nuclei (acellular Comet assay) of catalase deficient (CAT1AS) and wild-type (SR1) tobacco with the promutagen o phenylenediamine (o-PDA) and the direct acting genotoxic agents hydrogen peroxide and ethyl methanesulphonate (EMS). The roots of CAT1AS have about 60% less catalase activity compared to the roots of SR1. The promutagen o-PDA applied on tobacco roots induced significantly higher levels of DNA damage in the CAT1AS transgenic line than in SR1, while after application of o-PDA on isolated root nuclei, no DNA damage could be detected. In the catalase-deficient line CAT1AS about six-fold lower concentrations of H(2)O(2) are sufficient to induce the same levels of DNA damage as in SR1. By contrast, after treatment of isolated root nuclei with H(2)O(2) no difference in the induced levels of DNA damage was observed between CAT1AS and SR1. The DNA damaging effect of EMS was not affected by the presence of catalase in the tobacco roots and the levels of DNA damage measured by the cellular and acellular assay were similar. Comparing the effects of genotoxic agents in both the cellular and acellular Comet assays may help to elucidate their mechanism of action. Differences in both systems may reveal the participation of scavengers and of repair and metabolic enzymes on the activity of the genotoxic agent and the role of the cell wall in preventing the agent from reacting with nuclear DNA. PMID- 12581538 TI - Adaptive response to alkylating agents in the Drosophila sex-linked recessive lethal assay. AB - The adaptive response to alkylating agents was studied in Drosophila assays under various treatment procedures. Pre-treatment of males as well as treatment of females with low doses of EMS (0.05-0.1 mM) did not affect sex-linked recessive lethal (SLRL) rates induced by high doses of this mutagen (10 mM, various feeding duration) in mature sperm cells. Pre-treatment of males with a low dose of MMS (0.1 mM) enhanced mutagenesis induced by the high dose of EMS (10 mM) at different stages of spermatogenesis, the observed effects exceeding the additive action of both mutagens. On the contrary, larval pre-treatment with the adaptive dose of EMS (0.05 mM) resulted in resistance of their germ cells to higher doses of EMS (1 mM). Specifically, offspring production increased while dominant lethality in F(1) as well SLRL frequency in F(2) was significantly reduced as compared with the effects of larval exposure to the challenge dose. Under the conditions tested, the adaptive response of germ cells to alkylating agents was demonstrated in larvae, but not in adult flies. PMID- 12581540 TI - The dose-response relationship at very low doses of acrylamide is linear in the flow cytometer-based mouse micronucleus assay. AB - Acrylamide (AA) is genotoxic and has been classified as a probable human carcinogen. Human exposure to AA may be high by the consumption of starch-based food that has been treated at high temperature, e.g. potato chips and crisps. For risk assessment, extrapolation to the expected low doses to humans will be more reliable when data from low experimental doses can be used. We have registered the effects of a series of low doses in the sensitive flow cytometer-based micronucleus assay in mice, paying special attention to deviations from the expected linear dose-response function. Two experiments were performed with CBA mice, injected i.p. with different doses of AA. In one experiment the effects of 22 doses (two mice per dose) ranging from 0 to 100 mg/kg b.w. were studied. In the second experiment seven doses (five mice per dose) ranging from 0 to 30 mg/kg b.w. were used. In both experiments, a clear increase of the frequency of micronucleated erythrocytes was seen, already at the lowest doses used. The dose response function was found to be linear with a tendency to have a steeper rise at the lowest doses. The low DNA content of the micronuclei indicated an absence of whole chromosomes, i.e. no aneugenic effect of AA. PMID- 12581539 TI - Sister chromatid exchanges in lymphocytes of nuclear medicine physicians. AB - OBJECTIVE: The aim of this study was to assess whether occupational exposure to chronic, low doses of Iodine 131 (I-131) and Technetium 99m (Tc-99m) may lead to genotoxicity. Medical personnel occupied in nuclear medicine departments are occupationally exposed to low doses of I-131 and Tc-99m. The determination of the frequency of sister chromatid exchanges (SCEs) and of cells with a high frequency of SCEs (HFC) is considered to be a sensitive indicator for detecting genotoxic potential of mutagenic and carcinogenic agents. Therefore, we examined peripheral lymphocytes from nuclear medicine physicians for the presence of both SCE and HFC. METHODS: Sixteen exposed nuclear medicine physicians (non-smokers) were compared to 16 physicians (non-smokers) who had not been exposed to chemical or physical mutagens in their usual working environment at the same hospital. RESULTS: A statistically significant difference was found between SCE frequencies and HFC percentages measured in lymphocytes from the exposed and control groups. CONCLUSIONS: The present observation on the effect of chronic low doses of I-131 and Tc-99m indicates the possibility of genotoxic implications of this type of occupational exposure. Hence, the personnel who work in nuclear medicine departments should carefully apply the radiation protection procedures and should minimize, as low as possible, radiation exposure to avoid possible genotoxic effects. PMID- 12581541 TI - The seasonality of rhinovirus infections and its implications for clinical recognition. AB - BACKGROUND: Rhinoviruses are the most common cause of acute respiratory infections. Isolation of rhinoviruses occurs in a distinct and consistent seasonal pattern that can be used to help determine whether an acute respiratory illness is caused by a rhinovirus. OBJECTIVE: This article reviews information on the seasonality of rhinovirus infection derived from early and recent studies of rhinovirus occurrence and treatment. METHODS: PubMed was searched from 1965 to the present to identify all potentially relevant papers. The search terms used were rhinovirus and seasonality. A total of 1998 papers were screened. RESULTS: Rhinoviruses comprise more than three quarters of viruses circulating in early autumn. In some years and perhaps some geographic areas, spring is an even more important time for rhinovirus transmission. Although overall rates of respiratory illness are lower in summer, rhinoviruses are the most frequently isolated virus at this time of year. Other viral agents, including influenza viruses and respiratory syncytial virus (particularly with parainfluenza virus), predominate in the winter. Thus, for most of the year, rhinoviruses are the cause of the majority of acute viral respiratory infections. CONCLUSION: Understanding the seasonal incidence of rhinovirus infection may help determine how best to employ currently available antirhinoviral agents in patients presenting with symptoms of an acute viral respiratory infection. PMID- 12581542 TI - A comparison of the mechanisms of action of interferon beta and glatiramer acetate in the treatment of multiple sclerosis. AB - BACKGROUND: The development of immunomodulatory agents has represented a major advance in the treatment of multiple sclerosis (MS). To date, immunomodulatory agents approved for the treatment of relapsing MS in the United States include 3 forms of recombinant interferon (IFN) beta (2 formulations of IFN beta-1a and 1 of IFN beta-1b) and synthetic glatiramer acetate (GA). Recognition of how these agents work to regulate the immune system may lead to a better understanding of disease mechanisms, as well as to development of more effective therapies or combinations of therapy. OBJECTIVE: This article reviews the potential mechanisms of action of IFN beta products and GA in the context of their regulatory effects on autoimmune components that may be of importance in MS. METHODS: MEDLINE and Current Contents/Clinical Medicine were searched for articles published in English from 1993 to the present using the search terms interferon beta, glatiramer acetate, and multiple sclerosis. RESULTS: IFN beta products affect the disease process in MS through multiple potential mechanisms of action, including antiviral, antiproliferative, and anti-inflammatory effects. The mechanisms of action of GA are less clear, but may involve immune regulation induced by a gradual shift of T-cell phenotype from proinflammatory (type 1 T-helper cells) to anti-inflammatory (type 2 T-helper cells) and interference with antigen presentation. CONCLUSION: Understanding the mechanisms of action of IFN beta products and GA provides important insights into the disease processes involved in MS. PMID- 12581543 TI - Fenofibrate in the treatment of dyslipidemia: a review of the data as they relate to the new suprabioavailable tablet formulation. AB - BACKGROUND: The fibric acid derivative fenofibrate is indicated as an adjunct to dietary modification in adults with primary hypercholesterolemia or mixed dyslipidemia (types IIa and IIb hyperlipidemia, Fredrickson classification) to reduce levels of low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triglycerides (TG), and apolipoprotein (apo) B, and to increase levels of high-density lipoprotein cholesterol (HDL-C) and apo A. It is also indicated as adjunctive therapy to diet for the treatment of hypertriglyceridemia (types IV and V hyperlipidemia). Initially approved in the United States in a micronized capsule formulation, fenofibrate is now available in a new "suprabioavailable" tablet formulation that has increased bioavailability, achieving equivalent plasma concentrations at lower doses. The 67- and 200-mg micronized capsules can be considered equivalent to the 54- and 160-mg suprabioavailable tablets, respectively. OBJECTIVE: This paper reviews the pharmacologic properties, clinical usefulness, and safety profile of fenofibrate in the management of dyslipidemias. METHODS: Recent studies, abstracts, reviews, and consensus statements published in the English-language literature were identified through searches of MEDLINE (1966-January 2002), International Pharmaceutical Abstracts (1970-January 2002), and PharmaProjects (1990-January 2002) using the search terms fenofibrate, fibrates, hyperlipidemia, hypertriglyceridemia, and dyslipidemia. RESULTS: Fenofibrate is well absorbed after oral administration, with peak plasma levels attained in 6 to 8 hours. The absolute bioavailability of fenofibrate cannot be determined due to its being virtually insoluble in aqueous media suitable for injection; however, after oral administration of a single dose of radiolabeled fenofibrate, approximately 60% of the dose appeared in urine, primarily as fenofibric acid and its glucuronated conjugate, and approximately 25% was excreted in the feces. The apparent volume of distribution is 0.89 L/kg in healthy volunteers, and protein binding is approximately 99% in healthy and hyperlipidemic patients. Neither fenofibrate nor fenofibric acid appears to undergo significant oxidative metabolism in vivo. Fenofibric acid has a half-life of 20 hours. Fenofibrate is effective in lowering TG levels and increasing HDL-C levels. Its LDL-C-lowering effect is greater than that of gemfibrozil. Adverse effects of fenofibrate appear to be similar to those of other fibrates, including gastrointestinal symptoms, cholelithiasis, hepatitis, myositis, and rash. Fenofibrate therapy has been associated with increases in serum aminotransferase levels, and clinical monitoring of these markers of liver function should be performed regularly. CONCLUSIONS: Fenofibrate is effective in reducing levels of TG, TC, and LDL-C, and increasing levels of HDL-C in patients with dyslipidemias. Its efficacy and tolerability in the treatment of hypertriglyceridemia and combined hyperlipidemia have been demonstrated in numerous clinical trials. Its use is accompanied by a low incidence of adverse effects and laboratory abnormalities. Fenofibrate protects against coronary heart disease not only through its effects on lipid parameters but also by producing alterations in LDL structure and, possibly, alterations in the various hemostatic parameters. Its uricosuric property may prove to be a useful adjunctive attribute. PMID- 12581544 TI - A 12-month, multicenter, prospective, open-label trial of radiographic analysis of disease progression in osteoarthritis of the knee or hip in patients receiving celecoxib. AB - BACKGROUND: Studies have suggested that nonspecific nonsteroidal anti inflammatory drugs may inhibit matrix biosynthesis by articular cartilage, thereby accelerating the progression of osteoarthritis (OA). OBJECTIVE: The objective of this analysis was to determine whether 1-year treatment with the cyclooxygenase-2-specific inhibitor celecoxib at up to twice the recommended and maximally effective dose for OA had any deleterious effects on OA progression by assessing radiographic changes in knee or hip joint morphology in patients with OA. METHODS: In a 12-month, multicenter, prospective, open-label trial, patients with OA of the knee or hip or rheumatoid arthritis received celecoxib at doses ranging from that recommended for the treatment of OA (200 mg/d) to twice the recommended daily dosage (400 mg/d). Available radiographs showing baseline and end-of-treatment status were analyzed using semiquantitative measures of index joint morphology in patients with mild to moderate OA. The morphologic scores were then subjected to mean change and shift-table analysis to determine the extent and rate of disease progression. RESULTS: A total of 2,327 patients (796 with OA of the knee, 1,531 with OA of the hip) were included. A subset of 344 patients (160 with OA of the knee, 184 with OA of the hip) had radiographs from both before and after 12 months' celecoxib treatment. One hundred forty-seven and 158 pairs of knee and hip radiographs, respectively, were available for analysis. These revealed that, with the exception of significant hip joint-space narrowing (P = 0.029), no evidence of disease progression with long-term celecoxib treatment could be detected. The observed increase in hip joint-space narrowing was small (0.14 units/y) (95% CI, 0.08-0.20), was observed prior to celecoxib exposure (by mean change or shift-table analysis), and was not dose related. CONCLUSION: These results are consistent with the hypothesis that long-term therapy with celecoxib does not accelerate progression of OA of the knee or hip. PMID- 12581545 TI - Prospective, noninterventional, uncontrolled, open-chart, pharmacoepidemiologic study of prescribing patterns for lipid-lowering drugs at a tertiary care teaching hospital in North India. AB - BACKGROUND: The guidelines for management of dyslipidemia released by the US National Cholesterol Education Program (NCEP) have been questioned for their relevance in the South Asian Indian populations because these populations are reported to have significantly different lipoprotein parameters and atherogenic risk factors than Western populations. OBJECTIVE: The aim of this study was to determine current prescribing patterns for lipid-lowering drugs (LLDs) adopted by physicians in North India. METHODS: This prospective, noninterventional, uncontrolled, open-chart, pharmacoepidemiologic study was conducted from June 2000 to August 2000 at a tertiary care hospital in North India and included 200 dyslipidemic patients. The pattern of prescribing LLDs was recorded, along with the serum levels of lipid parameters-total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and lipoprotein(a) (Lp[a])-at the time of initiating LLD therapy and compared with the 1993 NCEP-II therapeutic guidelines for dyslipidemia management. RESULTS: The mean (SD) levels of lipid parameters in the study population were as follows: TC, 223.2 (21.5) mg/dL; TG, 258.4 (61.3) mg/dL; LDL C, 131.6 (26.5) mg/dL; HDL-C, 39.8 (8.9) mg/dL; and Lp(a), 44.8 (26.8) mg/dL. The LLDs prescribed were fibrates (53.5%) and statins (46.5%). Forty percent of patients prescribed LLDs did not meet the NCEP-II criteria for initiation of LLD therapy. CONCLUSIONS: Considerable differences in prescribing patterns of LLDs were observed compared with the then-prevalent NCEP-II guidelines. However, due to the abnormally high serum Lp(a) levels present in the average dyslipidemia profile in South Asian Indian populations, this pattern was in accordance with the specific recommendations made for these populations, as well as with the 2001 NCEP-III guidelines. PMID- 12581546 TI - A randomized, double-blind, single-dose, crossover clinical trial of the onset and duration of protection from exercise-induced bronchoconstriction by formoterol and albuterol. AB - BACKGROUND: Inhaled short-acting beta(2)-adrenoceptor agonists are the most commonly used treatment for the prevention of exercise-induced bronchoconstriction (EIB). Formoterol, a long-acting beta(2)-adrenoceptor agonist, has been demonstrated to provide protection from EIB, although the onset and duration of this protection have not been defined. OBJECTIVE: The purpose of this study was to determine the onset and duration of the protective effect of a single dose of inhaled formoterol powder against EIB, comparing them with the effect of a single dose of placebo and albuterol administered via metered-dose inhaler (MDI). METHODS: In this double-dummy, 4-way crossover study, patients received single doses of formoterol (12 and 24 microg) via a powder inhaler, albuterol by MDI (180 microg), and placebo. Exercise challenge tests (ECTs) were conducted at 15 minutes and at 4, 8, and 12 hours postdose. Pulmonary function studies (forced expiratory volume in 1 second [FEV(1)] and peak expiratory flow rate) were performed before and after each exercise challenge. RESULTS: Twenty adolescent and adult patients (mean age, 23.8 years; range, 13-41 years; 9 male, 11 female) with asthma were enrolled in the study, and 17 completed all 4 treatment sequences. Compared with placebo, both doses of formoterol produced significantly greater inhibition of FEV(1) decreases at all time points (P < 0.01). There were no significant differences in efficacy measures between the 2 formoterol doses throughout the study. The exercise-induced decrease in FEV(1) after albuterol treatment was significantly reduced compared with placebo only at 15 minutes after dosing (P < 0.05). Formoterol and albuterol exhibited a similar rapid onset of action (<15 minutes), but formoterol continued to protect patients against EIB for at least 12 hours (P < 0.01), whereas albuterol was no longer clinically effective by the 4-hour ECT. CONCLUSIONS: Formoterol and albuterol, given as single-dose inhalations, both provided protection from EIB within 15 minutes in this group of patients. The bronchoprotection afforded by formoterol lasted up to 12 hours, whereas that of albuterol was no longer significant by 4 hours. PMID- 12581548 TI - An open-label, randomized, multicenter, comparative study of the efficacy and safety of 7 days of treatment with clarithromycin extended-release tablets versus clarithromycin immediate-release tablets for the treatment of patients with acute bacterial exacerbation of chronic bronchitis. AB - OBJECTIVE: The aim of this study was to compare the efficacy and safety of clarithromycin extended-release (ER) tablets and immediate-release (IR) tablets. METHODS: This was a Phase III, open-label, randomized, multicenter, comparative study in ambulatory patients with a diagnosis of acute exacerbation of chronic bronchitis (AECB). Eligible patients were randomized 1:1 to receive either 1 clarithromycin ER 500-mg tablet QD for 7 days or 1 clarithromycin IR 250-mg tablet BID for 7 days. Clinical and bacteriologic responses were assessed within 48 hours after the last dose of study drug and at a test-of-cure visit 21 +/- 2 days posttreatment. RESULTS: Of 233 patients randomized, 162 (86/117 [73.5%] in the ER group and 76/115 [66.1%] in the IR group) completed the study protocol. Compliance did not differ significantly between the treatment groups; however, significantly fewer patients in the ER group reported missing doses of study medication than in the IR group (7/118 [5.9%] vs 16/115 [13.9%]; P = 0.04). The clinical cure rates for the clarithromycin ER and IR groups were 81.0% (68/84) and 82.1% (64/78) and the clinical success (clinical cure plus clinical improvement) rates were 94.0% [79/84] and 89.7% [70/78], respectively. There were insufficient data for analysis of bacteriologic efficacy. However, bacteria were eradicated or presumed eradicated in 71.4% (10/14) and 79.2% (19/24) of patients in the ER and IR groups, respectively. The number of adverse events (AEs) considered to be possibly or probably related to study drug (23.4% [52/222] of patients receiving clarithromycin ER and 24.4% [43/176] of patients receiving clarithromycin IR) was similar between groups, as was the severity of these events (94.2% [49/52] in the ER group classified as mild or moderate vs 93.0% [40/43] in the IR group). Overall, the most commonly reported AEs were diarrhea, nausea, abdominal pain, headache, and taste disturbance. CONCLUSION: Clarithromycin ER 500-mg tablets QD for 7 days were as effective and well tolerated as clarithromycin IR 250-mg tablets BID for 7 days in treating adults with AECB. PMID- 12581547 TI - Comparison of once-daily extended-release ciprofloxacin and conventional twice daily ciprofloxacin for the treatment of uncomplicated urinary tract infection in women. AB - BACKGROUND: Trimethoprim/sulfamethoxazole (TMP/SMX) is currently the first choice for empiric therapy of acute uncomplicated urinary tract infection (UTI) in women. In areas where resistance to TMP/SMX is known to be high, ciprofloxacin and other fluoroquinolones are recommended as first-line choices for the empiric therapy of UTI. OBJECTIVE: This study compared the efficacy and safety profile of once-daily extended-release ciprofloxacin 500 mg (referred to hereafter as ciprofloxacin QD) with those of conventional ciprofloxacin 250 mg BID, each administered orally for 3 days, in the treatment of uncomplicated UTI in women. METHODS: In this multicenter, prospective, randomized, double-blind, double dummy, Phase III trial, adult women with clinical signs and symptoms of acute uncomplicated UTI, pyuria, and a positive pretherapy urine culture (>/=10(5) colony-forming units/mL) received ciprofloxacin QD or ciprofloxacin BID. Bacteriologic and clinical outcomes were assessed at the test-of-cure visit (4-11 days after completion of therapy) and the late follow-up visit (25-50 days after completion of therapy). RESULTS: The intent-to-treat population consisted of 891 patients (444 ciprofloxacin QD, 447 ciprofloxacin BID); 422 patients were evaluable for efficacy (199 ciprofloxacin QD, 223 ciprofloxacin BID). At the test of-cure visit, bacteriologic eradication was achieved in 94.5% (188/199) of the ciprofloxacin QD group and 93.7% (209/223) of the ciprofloxacin BID group (95% CI, -3.5 to 5.1). Clinical cure was achieved in 95.5% (189/198) of the ciprofloxacin QD group and 92.7% (204/220) of the ciprofloxacin BID group (95% CI, -1.6 to 7.1). Bacteriologic and clinical outcomes at the late follow-up visit were consistent with the test-of-cure findings. The rate of eradication of Escherichia coli, the most prevalent organism, was >97% in each treatment group. Rates of drug-related adverse events were similar with the once- and twice-daily ciprofloxacin regimens (10% and 9%, respectively). CONCLUSION: Extended-release ciprofloxacin 500 mg given once daily for 3 days was as effective and well tolerated as conventional ciprofloxacin 250 mg given twice daily for 3 days in the treatment of acute uncomplicated UTI in women. PMID- 12581550 TI - An examination of the effect of cytochrome P450 drug interactions of hydroxymethylglutaryl-coenzyme A reductase inhibitors on health care utilization: a Canadian population-based study. AB - BACKGROUND: Cytochrome P450-related drug interactions can lead to adverse effects that may affect health care resource utilization. OBJECTIVE: The purpose of this study was to quantify the impact of drug interactions involving hydroxymethylglutaryl-coenzyme A reductase inhibitors (statins) on health care resource utilization. METHODS: Using the Manitoba Health Research database, we identified patients who had used statins between January 1, 1995, and March 31, 1998. New statin users (NSUs) were those who received a first prescription for a statin after April 30, 1995; old statin users (OSUs) were those who had a statin prescription before January 1, 1995. The number of hospitalizations, physician visits, and prescriptions, and their associated costs to the Manitoba health care system were calculated. Statin interacters were defined as users with >1 prescription for an interacting drug while receiving a statin. Interacting drugs were classified into 2 groups: group A included drugs whose levels increased as a result of the statin prescription; drugs in group B increased statin levels. The Wilcoxon rank-sum test was used to analyze differences by statin on health care resource use. RESULTS: A total of 28,705 statin users (18, 181 NSUs and 10,524 OSUs) were identified. During the study period, 24,496 (85.3%) individuals took 1 statin, 3751 (13.1%) took 2 statins, and 458 (1.6%) took 3 to 5 statins. The most common coadministered group A interacting drugs were diclofenac (5.8%), amitriptyline (4.9%), warfarin (4.5%), and ibuprofen (1.8%). The most common group B interacting drugs were erythromycin (8.2%), omeprazole (5.5%), cimetidine (3.6%), and clarithromycin (3.5%). Statin interacters consumed significantly more health care resources than did noninteracters for both incident and prevalent analyses (P < 0.001). In the prevalent analysis (NSUs + OSUs), pravastatin users taking interacting drugs had significantly fewer hospitalizations (mean, 1.3), fewer physician visits (mean, 24.2), and lower health care costs (mean, 5,526 dollars) compared with prevalent users of lovastatin (1.7, 28.0, and 6,925 dollars, respectively) and fewer physician visits than simvastatin users (25.6, P < 0.001). In the incident analysis, pravastatin users had significantly less physician visits (20.8 vs 23.5, P < 0.001) and lower health care costs (4,739 dollars vs 6,323 dollars, P < 0.001) than lovastatin users. CONCLUSION: Pravastatin was associated with fewer hospitalizations, physician visits, and overall health care resource utilization in prevalent users than lovastatin, possibly due to a lack of drug interaction effects. PMID- 12581551 TI - Validation of the short form of the Spanish Hypertension Quality of Life Questionnaire (MINICHAL). AB - BACKGROUND: The finding that low health-related quality of life (HRQoL) can be a risk factor for future cardiovascular events in patients with hypertension emphasizes the need for disease-specific HRQoL assessment tools in this population. OBJECTIVE: This study evaluated the feasibility and psychometric properties of the short form of the Spanish Hypertension Quality of Life Questionnaire (MINICHAL). METHODS: An observational, prospective, multicenter study was carried out among patients with hypertension, stratified by sex and disease stage. The feasibility and validity of the MINICHAL were evaluated at baseline. Two follow-up visits were conducted at 1 and 6 months after the initial visit, during which the test-retest reliability and responsiveness to change of the MINICHAL were assessed, respectively. RESULTS: A total of 736 consecutive patients with hypertension were included. Mean (SD) administration time for the MINICHAL was 7.2 (6.5) minutes. In the validation process, factor analysis showed 2 dimensions: "State of mind" (StM) and "Somatic manifestations" (SM). The response level was high, with 94.3% of patients answering all items. Patients with a higher disease stage (ie, stage II/III hypertension) had a worse HRQoL than those with a lower disease stage (P < 0.01). Internal consistency was 0.87 for StM and 0.75 for SM. Test-retest reliability was 0.82 and 0.75 for the StM and SM dimensions, respectively. Among patients who reported experiencing a positive change in their overall health status 6 months after treatment initiation or treatment change, responsiveness to change ranged from 0.52 (better) to 1.14 (much better). CONCLUSIONS: The MINICHAL questionnaire could be administered in a short time with a high response level and was found to have acceptable psychometric properties. The MINICHAL is a potentially useful tool for the evaluation of HRQoL among patients with hypertension in the routine clinical setting. PMID- 12581552 TI - Assessment of the mean-value of 17-hydroxypregnenolone in the umbilical blood of newborns by the exploratory analysis of biochemical data. AB - The main aim of data analysis in biochemical metrology is the extraction of relevant information from biochemical data measurements. A system of extended exploratory data analysis (EDA) based on the concept of graphical tools for sample data summarization and exploration is proposed and the original EDA algorithm in S-Plus is available on the Internet at http://www.trilobyte.cz/EDA. To check basic assumptions about biochemical and medical data is to examine the independence of sample elements, sample normality and homogeneity. The exact assessment of the mean-value and the variance of steroid levels in controls is necessary for the correct assessment of the samples from patients. Data examination procedures are illustrated by a determination of the mean-value of 17 hydroxypregnenolone in the umbilical blood of newborns. For an asymmetric, strongly skewed sample distribution corrupted with outliers the best estimate of location seems to be the median. The Box-Cox transformation improves a sample symmetry. The proposed procedure gives reliable estimates of a mean-value for an asymmetric distribution of 17-hydroxypregnenolone when the arithmetic mean can not be used. PMID- 12581553 TI - Improved prediction-based ovarian follicle detection from a sequence of ultrasound images. AB - A new algorithm is presented for ovarian follicle recognition from a sequence of ultrasound images. The basic version of the prediction-based algorithm is upgraded by means of two improvements. The negative influence brought by the gross measurement errors is suppressed, and the locality of the treated process is considered. The basis for both improvements is the Kalman filter. The proposed algorithm is a combination of three mutually dependent Kalman filters: a global one whose parameters are then modified by two additional ones, firstly detecting the gross measurement errors and secondly, regarding the recognised contour of the object. The obtained results show that the follicles recognised using the final prediction algorithm are about 2% more compact and about 6% more accurate, on average, when compared to the values obtained using the basic prediction-based algorithm. PMID- 12581554 TI - Pre-operative planning and gait analysis of total hip replacement following hip fusion. AB - A computer-based pre- and post-operative analysis of total hip replacement (THR) is presented. The analysis was performed by using and innovative combination of X ray measurements and gait analysis. The several important biomechanical factors that affect effective THR are analysed in a test clinical case of conversion of hip fusion into THR. Geometrical positioning of the artificial joint should restore correct motion and adequate leverage of the surrounding muscles in order to exert the appropriate functional joint moments. Hip joint centre positioning pre- and post-operation was evaluated using a computer-based X-ray measurement tool. This enabled the calculation of the medio-lateral and proximal-distal off set of the joint centre, the lever arms of the abductor muscles and the limb length discrepancy. Pre- and post-operative function was also evaluated using gait analysis, including joint kinematics, kinetics and electromyography. The whole analysis suggests that a good restoration of joint motion, a regression of pelvic compensation and a phasic activity of gluteus medius occur only when a good positioning of the prosthetic hip joint is obtained. This study proved a fundamental role played by these two computer-based tools in the support of the clinical decision making process. PMID- 12581555 TI - New method for simultaneous anatomical and functional studies of articular joints and its application to the human knee. AB - In this paper a new methodology is described for the acquisition and computer elaboration of joint anatomy and motion data and the study of their correlation. The method uses a particular commercial electrogoniometer never used before for biomechanic investigations and software developed by the author for numerical interpolations and interactive display of the anatomical structures during joint motion. The original data acquisition protocol and computer elaboration software are described in detail, the theoretical and experimental accuracy of the method is reported and the original features and potential benefits of this approach in the field of joint biomechanics are discussed. Finally an experimental application for geometrical and functional analysis of the knee is presented as a practical example. PMID- 12581556 TI - An integrated research tool for X-ray imaging simulation. AB - This paper presents a software simulation package of the entire X-ray projection radiography process including beam generation, absorber structure and composition, irradiation set up, radiation transport through the absorbing medium, image formation and dose calculation. Phantoms are created as composite objects from geometrical or voxelized primitives and can be subjected to simulated irradiation process. The acquired projection images represent the two dimensional spatial distribution of the energy absorbed in the detector and are formed at any geometry, taking into account energy spectrum, beam geometry and detector response. This software tool is the evolution of a previously presented system, with new functionalities, user interface and an expanded range of applications. This has been achieved mainly by the use of combinatorial geometry for phantom design and the implementation of a Monte Carlo code for the simulation of the radiation interaction at the absorber and the detector. PMID- 12581557 TI - Delivery of medical multimedia contents through the TCP/IP network using RealSystem. AB - We developed a low cost, user-friendly multimedia delivery system, to provide medical lectures saved as multimedia contents to persons engaged in medicine. This system was created using the RealSystem package with the TCP/IP network. Users can review lectures and medical meeting presentations with video and audio through the Internet, whenever convenient. Each medical source of video and slide has been clearly displayed on a screen. Members of medical associations or medical students can easily review the most interesting parts of these files. This system is being used efficiently in distance learning and aids the diffusion of the latest information and technology to busy physicians and medical students. PMID- 12581558 TI - Solutions to the Behrens-Fisher problem. AB - When testing the equality of the means from two independent normally distributed populations given that the variances of the two populations are unknown but assumed equal the classical Student's two sample t-test is recommended. If the underlying population distributions are normal with unequal and unknown variances, either Welch's t-statistic or Satterthwaite's Approximate F test is suggested. However, Welch's procedure is non-robust under most non-normal distributions. There is a variable tolerance level around the strict assumptions of data independence, homogeneity of variances, identically and normal distributions. Few textbooks offer alternatives when one or more of the underlying assumptions are not defensible. We have developed an executable FORTRAN code for producing the statistics suggested by Cressie and Whitford, Yuen and Dixon, and Yuen. An executable FORTRAN is available from the author on request (e-mail only). PMID- 12581559 TI - Token swap test revisited. AB - The token swap test measures the association between row and column variables of a 2 x 2 table in sample misclassification space, and makes no assumptions about repeated, random sampling from a source population. Despite its conceptual usefulness, the token swap test is not implemented by standard statistical software packages. Here the author describes 'tokenSwaps', a Mathematica program that performs a token swap test. The 'tokenSwaps' program also performs a one tailed Fisher exact test, allowing results of the two methods to be compared. The program uses recursive functional programming and local rewrite rules to achieve substantial coding economy. Examples of the operation of the program are given, and its limitations are discussed. PMID- 12581560 TI - Development of computerized Kana Pick-out Test for the neuropsychological examination. AB - The Kana Pick-out Test, which was developed in Japan and done with paper and pencil, is said to be suitable for inspecting higher-order brain function and to be a good method for screening persons with mild or slight dementia. We have developed a computerized version of the Kana Pick-out Test, which runs on a stand alone computer, intended to be utilized for mass screening and self administration. The program was developed with Microsoft Visual Basic 6.0 and runs under the Windows operating system on any IBM PC compatible computer. In this study, all subjects could use the system by interacting with the computer and it was found that the system seemed to have the capability of detecting cognitive status equal to the paper-based Kana Pick-out Test. Besides this, we developed a network-based Kana Pick-out game software which was intended to attract user's notice. The game program was written in JAVA language and runs on a web-browser supporting JAVA on any operating system. The program, a so called applet, is located on our web site (http://environ.med.tottori-u.ac.jp) and anyone can use the applet by accessing our homepage. PMID- 12581561 TI - A global approach to tobacco policy. AB - The battle to control lung cancer has been an outcome of policies established over the years, largely since the 1960s. Early policies focussed on abolition of tobacco promotion, help in cessation of smoking, health warnings and labeling with tar and nicotine yields, public education, reduction of tar and nicotine yields, increases in tax and prevention of sales to minors. Over time the advent of nicotine replacement therapy, earmarking of tax for health purposes and counter-advertising became part of this policy. Over time the low tar policy delivered some but not all the benefits expected and the reasons for this are described. Modern approaches now feature regulation of the product with a reduction in the amounts of carcinogens and toxins in the smoke, a new measurement system for nicotine, a discussion of the options for nicotine policy and reduction of exposure to secondhand smoke. PMID- 12581562 TI - Is it more important to quit smoking than which chemotherapy is used? AB - Many patients continue to smoke after their diagnosis of cancer. It is a critically important time to work with the patient to have them stop smoking. Many oncologists may underestimate the potential interactions of smoking with their anti-tumor therapy. The detrimental possibilities are examined as they relate to surgery, radiation and most importantly, to chemotherapy. Oncologists must become more active in working with their continuing smokers in order to aid in their response to interventions. Documentation of smoking status is critical to report in future studies that have evaluated therapeutic options. PMID- 12581563 TI - Gelatinolytic activity of matrix metalloproteinase in lung cancer studied using film in situ zymography stamp method. AB - In this study, we investigated activity of matrix metalloproteinase (MMP) of lung cancer by newly developed film in situ zymography (FIZ) stamp method, which allows visual localization of gelatinolytic activity within the cut surface of a tumor. We performed FIZ stamp method and conventional gelatin zymography in 39 resected specimen of lung cancer. The degree of gelatinolytic activity was scored (FIZ score) and correlated with the clinicopathological factors of the tumor. FIZ score of normal lung was very low. Lung cancer tissue had consistently higher FIZ score than the matched normal lung tissue. There were statistically significant differences in the FIZ score according to the pathological stage (P = 0.0015), nodal status (P = 0.0007) and lymphatic invasion (P = 0.0004). Direct correlation was observed between the FIZ score and MMP-2 activity (rho = 0.568, P = 0.0030) as quantitated using conventional gelatin zymography. MMP-2 may play an important role in the lymphatic invasion of lung cancer. FIZ stamp method may be a simple and useful diagnostic aid for the presence of cancer cells in the resected specimen. PMID- 12581564 TI - Neuroendocrine and cytokeratin serum markers as prognostic determinants of small cell lung cancer. AB - This retrospective study aimed at determining the prognostic significance of neuroendocrine markers chromogranin A (CgA), pro-gastrin releasing peptide (ProGRP) and neuron-specific enolase (NSE), together with the cytokeratin 19 marker CYFRA 21-1 in small cell lung cancer (SCLC). A total of 148 histologically proven and previously untreated SCLC patients were included. Among them 118 patients received a cisplatin-etoposide combination or cisplatin-etoposide cyclophosphamide-4'-epidoxorubicin combination. All tumour markers were tested using immunoradiometric assays except for ProGRP which was tested using an enzyme linked immunosorbent assay. The thresholds for marker serum titrations were 53 pg/ml, 65, 17, and 3.6 ng/ml for ProGRP, CgA, NSE and CYFRA 21-1 respectively. Univariate analysis showed that patients affected by one of the following characteristics proved to have a significant shorter survival in comparison with the opposite status of each variable: age over 63 years, extensive-stage, serum LDH level higher than 600 U/l, serum NSE level higher than 17 ng/ml, serum CgA level higher than 65 ng/ml and serum CYFRA 21-1 level higher than 3.6 ng/ml. In addition, there was a trend towards a statistical significance for a high serum alkaline phosphatase level and a performance status equal to or worse than two. The following variables were independent determinants of a poor outcome: a poor performance status (hazard ratio [95% confidence interval]: 1.51 [1.02-2.22]), a high CgA level (HR: 1.61 [1.06-2.45]), a high CYFRA 21-1 level (HR: 2.10 [1.40 3.14]) and an age older than 63 years (HR: 1.68 [1.14-2.48]). When the multivariate analysis was restricted to patients receiving a cisplatin-etoposide based chemotherapy, the same variables were prognostic determinants with nearly similar hazard ratios. In conclusion, aside classical variables such as age and performance status, high serum CYFRA 21-1 and high serum CgA level in SCLC are both prognostic determinants of prognosis, in particular in patients receiving conventional chemotherapy consisting of cisplatin and etoposide-based combinations. PMID- 12581565 TI - Bcl-2 and bax expression in advanced non-small cell lung cancer: lack of correlation with chemotherapy response or survival in patients treated with docetaxel plus vinorelbine. AB - Surgical series of non-small cell lung cancer (NSCLC) pathologic samples have shown that the expression of the proteins bcl-2 and bax, which regulate cell death, may have prognostic implications. Laboratory data also suggests that these proteins may impact chemotherapy response. In order to determine the rate of bcl 2 and bax expression in advanced NSCLC and assess the impact on chemotherapy response and patient survival, we performed immunohistochemistry on biopsy samples from patients enrolled in a phase I/II trial of vinorelbine plus docetaxel. We chose to study the pathology of patients in this specific trial because both docetaxel and vinorelbine phosphorylate bcl-2 and we hypothesized that this mechanism may affect clinical outcome. The goal of this study was to determine the feasibility of this analysis, and to observe any differences in response rate or survival based on bcl-2 or bax staining results. Unstained slides from paraffin blocks were obtained for 31 patients (55%) on the phase I/II trial. The patient characteristics for this subgroup did not differ significantly from the entire cohort of patients on the trials. Bcl-2 staining was positive in 5/31 samples (16%, 95% CI 3-29%) and bax was positive in 19/28 samples (68%, 95% CI 51-85%). Bcl-2 and bax staining did not correlate with response (p = 0.65 and 1.00 respectively, Fisher's exact test), or survival (by Kaplan-Meier curves). In conclusion, bcl-2 and bax expression was similar in this population with advanced NSCLC to previously reported results for early stage disease, but did not predict response to vinorelbine plus docetaxel in this series. PMID- 12581566 TI - Monocyte-derived microparticles may be a sign of vascular complication in patients with lung cancer. AB - We measured and compared the levels of plasma monocyte-derived microparticles (MDMP) and platelet activation markers [plasma platelet-derived microparticles (PDMP), CD62P binding to platelets; plt-CD62P, CD63 binding to platelets; plt CD63], to develop a better understanding of their potential contribution to vascular complications of lung cancer. The concentrations of MDMP and PDMP in lung cancer patients were significantly higher (P < 0.01) than those in normal subjects. Levels of plt-CD62P and plt-CD63 were significantly higher (P < 0.001 for each) in lung cancer patients than in controls. Levels of sE-selectin were also higher in lung cancer patients than in control subjects. MDMP correlated positively with plt-CD62P, plt-CD63, and PDMP with its relation to PDMP being particularly significant. The number of MDMPs and PDMPs are patients who are non small cell lung cancer were significantly higher than that in small cell lung cancer patients. In addition, levels of sE-selectin were higher in non-small cell lung cancer than in small cell lung cancer patients. These findings suggest that elevated MDMPs may be a sign of vascular complication in lung cancer patients, particularly those who suffer from non-small cell lung cancer. PMID- 12581567 TI - The value of [18F]fluoro-2-deoxy-D-glucose positron emission tomography in the selection of patients with stage IIIA-N2 non-small cell lung cancer for combined modality treatment. AB - Combined modality treatment (CMT) for patients with stage IIIA-N2 non-small cell lung cancer (NSCLC) is at present studied extensively. To select patients with truly stage IIIA-N2 disease, however, proves to be difficult with current diagnostic tests. Distant metastases may become clinically overt during induction chemotherapy (IC) or shortly after, revealing the inaccuracies of current staging algorithms. A prospective study with [18F]fluoro-2-deoxy-D-glucose positron emission tomography (FDG PET) in IIIA-N2 NSCLC patients was performed to assess its value in the selection of this patient group. Fifty-seven patients received a whole body FDG PET scan as part of an ongoing response monitoring trial. Results were compared with conventional staging. In 32/57 (56%) PET suggested upstaging, which was confirmed in 17/57 (30%) with a median follow-up of 16 (range 2-49) months. These results show that using the conventional staging algorithm a substantial group of patients was understaged. FDG PET improves the selection of patients suitable for CMT. PMID- 12581568 TI - Correlation between lymph node micrometastasis and histologic classification of small lung adenocarcinomas, in considering the indication of limited surgery. AB - The validity of limited surgery for lung carcinomas have not been clearly established. Several reports suggest that not only tumor size but also the histological findings may be of importance in this role. By conducting immunohistochemical examinations of lymph node micrometastases from small lung adenocarcinomas, we considered the suitability of limited surgery for especially localized bronchioloalveolar carcinoma (LBAC) without active fibroblastic proliferation (Noguchi's histologic classification types A and B). In this study, we enrolled 54 patients with lung adenocarcinoma smaller than 2 cm in diameter. Lymph node involvement was demonstrated in 11 (20.4%) patients. Lymph node micrometastasis was present in 11 (25.6%) of the 43 patients at pN0 disease. However, 13 patients with LBAC without active fibroblastic proliferation (types A and B) had neither lymph node involvement nor micrometastasis. Our results indicate that limited pulmonary resection may be acceptable procedure for LBAC without active fibroblastic proliferation. PMID- 12581569 TI - Serum anti-p53 autoantibodies in pleural malignant mesothelioma, lung cancer and non-neoplastic lung diseases. AB - Alterations of the p53 gene may lead to the production of detectable autoantibodies (p53-Abs) in cancer patients. In order to evaluate the association of p53-Abs with pleuropulmonary diseases, four groups of subjects were analyzed by ELISA for serum p53-Abs, in the framework of a molecular epidemiologic study. Two of 30 pleural malignant mesothelioma patients (MM; 6.7%) and 8/48 lung cancer patients (LC; 16.7%) were seropositive, while all 51 healthy controls (HC) were negative. Two of 55 (3.6%) at-risk controls (RC) with non-malignant respiratory diseases were positive and were not subsequently diagnosed any cancer. The difference was statistically significant between LC and RC or HC (P = 0.01), but not between MM and any other group. No correlation was found with age, sex, cancer stage or histology, cigarette smoking or occupational exposure. A longer survival (not significant) was shown in seropositive LC but not in MM. p53 expression in tumor tissue was also evaluated in a subgroup of MM. In conclusion, the presence of detectable p53-Abs in serum was associated in a statistically significant proportion of cases with LC but only occasionally with MM. The longer survival among positive LC patients and the presence of two seropositive among patients with non-neoplastic respiratory diseases should be further investigated. PMID- 12581570 TI - Concomitant weekly docetaxel, cisplatin and radiation therapy in locally advanced non-small cell lung cancer: a dose finding study. AB - The optimal dose of weekly docetaxel in combination with cisplatin and concomitant thoracic radiation therapy (XRT) in patients with locally advanced non-small cell lung cancer (NSCLC) is not well defined. The purpose of this study was to define the maximal tolerated dose (MTD) of docetaxel in this combination. Eligible patients had unresectable stage IIIA or IIIB NSCLC without pleural effusion. Treatment consisted of cisplatin 25 mg/m(2) plus docetaxel weekly and concomitant standard XRT for a total of 60 Gy at 200 cGy/fraction/day 5 times weekly for 6 weeks. The starting dose of docetaxel in the first cohort was 15 mg/m(2)/week. This dose was escalated by 5 mg/m(2) per cohort of 3 patients. No intrapatient dose escalation was allowed. The doses of cisplatin and XRT were not escalated. A total of 23 patients were enrolled, and 19 patients were evaluable for analysis. The first cohort (docetaxel 15 mg/m(2)/week) completed treatment without any Grade 3 or 4 toxicities. The second cohort (docetaxel 20 mg/m(2)/week) was expanded to 6 patients because of Grade 3 cough observed in 1 patient. One of 5 patients experienced Grade 3 esophagitis at the docetaxel 25 mg/m(2)/week dose level. Dose limiting toxicity consisting of Grade 3 esophagitis was reached in 4 of 5 patients receiving docetaxel at 30 mg/m(2)/week. This study determined the MTD of weekly docetaxel to be 25 mg/m(2) when combined with cisplatin 25 mg/m(2) and radiation therapy for locally advanced NSCLC. Further evaluation of this regimen in a phase II trial is underway. PMID- 12581572 TI - A phase II study of bryostatin-1 and paclitaxel in patients with advanced non small cell lung cancer. AB - BACKGROUND: Bryostatin-1 is a macrocyclic lactone, which exhibits pleiotropic biological effects via protein kinase C and has shown preclinical synergy with paclitaxel for enhanced tumor cell apoptosis. PATIENTS AND METHODS: Patients had stage IIIB (pleural effusion)/IV non-small cell lung cancer, measurable disease, performance status 0-2 Eastern Cooperative Oncology Group, adequate organ function, and no prior chemotherapy. Patients received dexamethasone premedication followed by paclitaxel at a dose of 90 mg/m(2) on days 1, 8, and 15 along with bryostatin-1 50 microg/m(2) on days 2, 9, and 16 every 28 days until disease progression. Correlative assays measuring serum levels of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and T-lymphocyte numbers were performed based on a previous study showing cytokine induction in vivo by bryostatin-1. Fifteen patients were enrolled. RESULTS: Thirty cycles of the bryostatin-1 and paclitaxel were delivered with a median of 2 per patient (range 1-4). Myalgia was the predominant non-hematologic toxicity encountered as 3 patients developed grade 4 and 1 patient developed grade 3 myalgia. Four patients were removed from the study during cycle 1 for rapid disease progression or myalgia. Eleven patients could be evaluated for response. Five patients had stable disease, two had a mixed response, and four had progressive disease. Ten patients received second-line chemotherapy after leaving the study. Median survival was 31 weeks (95% confidence interval: 5.4-49.3). Correlative data showed a trend towards decreased plasma IL-6 and TNF-alpha after each cycle of therapy presumably due to the dexamethasone premedication and/or paclitaxel. CONCLUSIONS: This drug combination showed no significant clinical response and was associated with reproducible toxicity. The predominance of myalgia in the absence of elevated serum cytokines suggests a non-inflammatory etiology of this toxicity. PMID- 12581571 TI - Gemcitabine and cisplatin versus vinorelbine and cisplatin versus ifosfamide+gemcitabine followed by vinorelbine and cisplatin versus vinorelbine and cisplatin followed by ifosfamide and gemcitabine in stage IIIB-IV non small cell lung carcinoma: a prospective randomized phase III trial of the Gruppo Oncologico Italia Meridionale. AB - PURPOSE: we carried out a phase III randomized trial to compare vinorelbine cisplatin regimen to gemcitabine-cisplatin regimen, and to a sequential administration of gemcitabine-ifosfamide followed by vinorelbine-cisplatin or the opposite sequence of vinorelbine-cisplatin followed by ifosfamide-gemcitabine according to the 'worst drug rule' hypothesis in patients with locally advanced unresectable stage IIIB or metastatic stage IV non-small cell lung cancer. The primary endpoint was survival parameters, while secondary endpoints included analysis of response rates and toxicity. PATIENTS AND METHODS: patients were randomized to receive: (a) gemcitabine 1000 mg/m(2) on days 1, 8 and 15 plus ifosfamide 1500 mg/m(2) on days 8-12 with mesna uroprotection (GI regimen) followed by vinorelbine 25 mg/m(2) on days 1 and 8 plus cisplatin 100 mg/m(2) on day 1 (GI --> VC regimen); (b) the opposite sequence (VC --> GI); (c) vinorelbine plus cisplatin as above described (VC regimen); or (d) gemcitabine 1400 mg/m(2) on days 1 and 8 plus cisplatin 100 mg/m(2) on day 8 (GC regimen). All regimens were given every 4 weeks. All patients were chemotherapy naive and had a ECOG PS 0-2. RESULTS: 400 patients were enrolled into the trial. Interim analysis after inclusion of 243 patients showed that ORR were 19% in the GI --> VC arm, 32% in the inverse sequence arm (CV --> GI), 42% in the VC arm, and 30% in the GC arm. The VC arm was statistically superior over the GI --> VC arm (p = 0.0074), but not over the other regimens. Median TTP was 3.1 months in the GI --> VC arm versus 5.0 months in the VC --> GI arm (p = 0.014). For these reasons the GI --> VC and VC --> GI arm were closed since the 'worst drug rule' hypothesis was rejected. Accrual in the VC and GC arms continued up to 140 and 138 patients respectively. Final ORR were 44% for the VC regimen (4 CR), and 34% for the GC regimen (1 CR). This difference was statistically significant (p = 0.032). OS was 9.0 and 8.2 months, respectively, with no statistically significant difference. The 1-year survival rate was 24 and 20%, respectively for VC and GC regimens. As expected the incidence of phlebitis was higher in the VC arm, while thrombocytopenia, flu-like syndrome and asthenia were more frequent in the GC arm. CONCLUSIONS: the results of this trial indicate that the combination of vinorelbine and cisplatin and that of gemcitabine and cisplatin are equivalent in terms of median TTP and OS, although the vinorelbine-cisplatin regimen is associated with a higher ORR. Both regimens may be considered as reference treatments for future studies. Moreover, our data reject the 'worst drug rule' hypothesis of sequential treatments in NSCCL at least with the combination used in this study. PMID- 12581573 TI - Phase 2 study of cryptophycin 52 (LY355703) in patients previously treated with platinum based chemotherapy for advanced non-small cell lung cancer. AB - BACKGROUND: Cryptophycin 52 is a novel antitubulin drug with in vitro and in vivo activity in non-small cell lung cancer. Based upon promising Phase 1 data, a multicenter trial was performed to evaluate the drug in previously treated non small cell lung cancer (NSCLC). METHODS: Patients with Stage IIIb (pleural effusion) or Stage IV NSCLC and performance status 0-1 with adequate organ function who had received at least one and no more than two prior chemotherapy regimens (one of which must have contained a platinum agent) were eligible. Cryptophycin 52 was administered at a dose of 1.5 mg/m(2) day 1 and 8 every 3 weeks. Patients were reassessed every two cycles. RESULTS: Twenty-six patients were enrolled of whom 25 are evaluable for toxicity and response. There were no responders, toxicity was predominantly neurologic in the form of peripheral neuropathy and constipation. After the first 12 patients were enrolled, the dose was lowered to 1.125 mg/m(2) day 1 and 8. Toxicity was substantially reduced with this maneuver. Median survival was 4.1 months. The median number of cycles was two, however ten patients received four or more courses of therapy. CONCLUSION: Cryptophycin 52 failed to produce measurable responses utilizing this schedule. In 40% of patients there was evidence of disease stabilization. Toxicity at 1.5 mg/m(2) was unacceptable. Since activity and toxicity may be dose and schedule dependent, other schedules of cryptophycin 52 should be considered. PMID- 12581574 TI - Three-week schedule of irinotecan and cisplatin in advanced non-small cell lung cancer: a multicentre phase II study. AB - A phase II multicentre study of a 3-week schedule of irinotecan (CPT-11) and cisplatin providing the highest recommended dose intensity of both agents in combination, was conducted in patients with advanced non-small cell lung cancer (NSCLC). Seventy-four stage IIIB (not suitable for radiotherapy) or stage IV NSCLC patients were enrolled to receive CPT-11 200 mg/m(2) i.v. and cisplatin 80 mg/m(2) i.v. on day 1 every 3 weeks. Relative dose-intensities for CPT-11 and cisplatin were 92 and 95%, respectively. No complete responses were observed. Twenty-five patients out of 73 obtained a partial response (34.2%). Partial responses were confirmed in 18 patients (24.7%: 95% CI, 15.3-36.1%). Median survival overall was 8.2 months, 9.7 months for patients with baseline performance status (PS) 0 and 1, and 4 months for patients with PS 2. The 1-year survival rate was 31%. Major clinical toxicities were grade 3 and 4 delayed diarrhoea (29% of patients) and febrile neutropenia (14% of patients). In conclusion, the present once-every-3-week schedule of CPT-11 and cisplatin is feasible and active in PS 0-1 advanced NSCLC patients, but results do not seem superior to those reported with other schedules. PMID- 12581575 TI - Phase II study of docetaxel and ifosfamide combination chemotherapy in non-small cell lung cancer patients failing previous chemotherapy with or without paclitaxel. AB - Docetaxel has shown activity in the treatment of non-small-cell lung cancer (NSCLC) that has failed previous chemotherapy. Ifosfamide is an active alkylating agent used in the first-line treatment of NSCLC. We conducted a phase II study of docetaxel and ifosfamide chemotherapy in two groups (one with and one without previous paclitaxel treatment) of NSCLC patients who had failed previous chemotherapy, to assess the response and toxicity of this combination chemotherapy. Fifty patients were enrolled from June 2000 to December 2001, including 26 patients treated with paclitaxel-containing agents and 24 patients who had never been treated with paclitaxel. Treatment consisted of docetaxel 60 mg/m(2) and ifosfamide 3 g/m(2) intravenous infusion on day 1 of every 3 weeks. Two hundred and thirty-eight cycles of treatment were given, with a median of 5 cycles (range, 1-8 cycles). All patients were evaluable for toxicity profile and response rate. The major toxicity was myelosuppression. Grade 3 or 4 neutropenia occurred in 40 patients (80%) during treatment. Febrile neutropenia occurred in 7 patients (14%). Grade 3 anemia occurred in 2 patients. The majority of patients needed a decrease in the treatment dose due to grade 4 or febrile neutropenia. Interstitial pneumonitis occurred in 3 patients, leading to the death of two. Other toxicities were few and mild in severity. After two cycles of treatment, 5 patients (10%) had a partial response (95% confidence interval 1.7-18.3%), including 2 patients previously treated with paclitaxel and 3 who had not received this treatment. More patients who had been previously treated with paclitaxel suffered from progressive disease than among those who had never been treated with paclitaxel (P = 0.049). The median time to disease progression was 5 months and the median survival was 8.2 months. Median survival was 7.6 and 8.7 months, respectively, in patients with and without previous paclitaxel treatment (P = 0.56). Median survival was 8.7 and 7.6 months in patients receiving docetaxel and ifosfamide as second- and third-line chemotherapy, respectively (P = 0.327). In conclusion, docetaxel and ifosfamide salvage chemotherapy produces a relatively low response rate, low dose intensity, and higher proportion of severe neutropenia in NSCLC. Physicians should be alert to the potential of interstitial pneumonitis. Nevertheless, median survival was of a reasonable duration. PMID- 12581576 TI - A phase II multicenter study of combined topotecan and gemcitabine as first line chemotherapy for advanced non-small cell lung cancer. AB - This trial was designed to determine the 1-year survival rate, efficacy, and safety, produced by topotecan and gemcitabine as first line chemotherapy in advanced non-small cell lung cancer (ANSCLC). Fifty-three patients were enrolled; 51 received treatment. Topotecan 1 mg/m(2), days 1-5 and gemcitabine 1 g/m(2) days 1 and 15 were administered IV, each drug over 30 min; cycles consisted of 28 days. Treatment continued until progressive disease or intolerable toxicity. Nineteen patients (36%) had Eastern Cooperative Oncology Group criteria performance status (ECOG PS) = 0, 34 (64%) PS = 1. Median age was 64 years; 37 patients (70%) were male. HISTOLOGY: adenocarcinoma (42%), squamous cell carcinoma (28%), large cell (19%), and unclassified (11%). Among 47 evaluable patients, eight (17%) had partial response, 11 (23%) had stable disease. One-year survival was 39% and median survival was 7.6 months (range, < 1-19.6). Grade 3 and 4 toxicities included neutropenia (53%), anemia (18%), thrombocytopenia (12%), asthenia (8%), and gastrointestinal disorders (8%); three patients (6%) experienced neutropenic fever. There were no treatment-related deaths. The combination topotecan/gemcitabine produced a 1-year survival similar to previous platinum-based regimens, when used as first line chemotherapy for ANSCLC. The toxicity profile was acceptable. PMID- 12581577 TI - Phase I/II trial of vinorelbine and divided-dose carboplatin in advanced non small cell lung cancer. AB - Vinorelbine administered in a doublet with cisplatin has become a standard treatment in patients with advanced non-small cell lung cancer (NSCLC). However, carboplatin appears to provide comparable efficacy with a better nonhematologic safety profile than cisplatin. Herein we report the results of a phase I/II trial of weekly vinorelbine and divided-dose carboplatin in patients with stage IIIB/IV NSCLC, Eastern Cooperative Oncology Group performance status < or = 2, and adequate bone marrow. Patients received vinorelbine starting at 20 mg/m(2) (to 25 mg/m(2)) and carboplatin area under the curve (AUC) 2.5 in divided-doses, both given on Days 1 and 8 every 21-day cycle for up to 6 cycles or until disease progression. Dose-limiting toxicity was defined for Cycles 1 and 2. Tumor response and toxicity were assessed using standard criteria. Twenty-one patients with a mean age of 67 years (range, 43-79) and stage IIIB/IV (8/13) disease were enrolled. All but 1 patient were chemotherapy-nai;ve; the majority (n = 20) had good performance status (< or = 1). Seventy-nine courses (median, 4) were administered. The vinorelbine/carboplatin doublet was well tolerated, with 7 courses interrupted or delayed because of toxicity. Toxicities were generally mild and evenly divided between hematologic (i.e., neutropenia) and nonhematologic (i.e., fatigue). No growth factor support was required for hematologic toxicity. There was only one case of grade 2 alopecia, and no cases of > or = grade 2 neurotoxicity. There were 5 (24%) partial responses, and 9 (43%) patients had stable disease. Weekly vinorelbine 25 mg/m(2) and divided-dose carboplatin AUC 2.5 is a well tolerated regimen with activity in advanced NSCLC patients. Further evaluation of this regimen in combination with novel targeted biologic therapy is warranted. PMID- 12581578 TI - Endobronchial carcinoid tumor combined with pulmonary non-tuberculous mycobacterial infection: report of two cases. AB - We report here two cases of endobronchial carcinoid tumor complicated with pulmonary infection with non-tuberculous mycobacteria (NTM). Case 1 was an 81 year-old woman with the left lower lobe atelectasis. Bronchoscopy showed complete obstruction of the left basal bronchus by a tumor and a sleeve lower lobectomy with mediastinal lymph node dissection was performed. Pathological examination showed typical carcinoid located in the left basal bronchus and many caseous granulomas containing mycobacteria in the lung parenchyma distal to the bronchus. Bacterial examinations of sputum and gastric juice after the operation showed a growth of Mycobacterium kansasii. Case 2 was a 50-year-old woman with the atelectasis of the left upper division. Bronchoscopy showed complete obstruction of the left upper division bronchus by a tumor and a left upper lobectomy with mediastinal lymph node dissection was performed. Pathological examination showed typical carcinoid located in the left upper division bronchus and many caseous granulomas in the lung parenchyma distal to the bronchus. The Ziehl-Neelsen stain showed many mycobacteria in these granulomas and they were identified as Mycobacterium avium by PCR analysis. Although NTM are not well recognized as possible pathogens of pulmonary infection related to bronchial obstruction by endobronchial carcinoma, our experiences rouse a caution to consider NTM as potential pathogens. We also discuss the possible mechanisms responsible for the specific relationship between carcinoid tumor and TNM. PMID- 12581579 TI - Early lung cancer action project pathology protocol. PMID- 12581580 TI - Assessment of antibody avidity in aborting cattle by a somatic Neospora caninum tachyzoite antigen IgG avidity ELISA. AB - A Neospora caninum IgG avidity ELISA was carried out on the basis of a somatic N. caninum tachyzoite antigen. The test was validated using experimentally infected calves, where a clear maturation of the IgG avidity over time could be demonstrated. At a maximum of 82 days after infection (d.p.i.), all animals showed antibody avidities ranging above 35%, and respective sera were thus defined as highly avid. Sera of 103 naturally infected seropositive cows with abortion (N. caninum association was provided by a N. caninum PCR-positivity of the fetus in 40 cases) and 139 seropositive animals without abortion history were concurrently examined. Significantly lower avidities were observed in aborting cows when compared to animals without abortion problems (P<0.01). While the avidity of sera collected before abortion remained practically constant until abortion, a significant increase of avidity could be observed in samples collected weeks to months after abortion (P<0.01). The avidities of non-aborting animals from farms with or without abortion problems did not differ significantly with time and were mainly located in the high avidity area. These data indicate that low avidities are not necessarily linked to recent N. caninum infection but can also be an indicator for increased abortion risk in cattle. PMID- 12581581 TI - Sequence comparison of Sarcocystis neurona surface antigen from multiple isolates. AB - Sarcocystis neurona is responsible for equine protozoal myeloencephalitis (EPM) and is phylogenetically closely related to Toxoplasma gondii and Neospora spp. There has been major progress in evaluating immunodominant and surface proteins of T. gondii and Neospora spp.; however, there is a paucity of data on the proteins or genes from S. neurona and S. neurona-like parasites. In this study, using reverse transcription-PCR (RT-PCR) and restriction fragment length polymorphism (RFLP) methods, the surface antigen gene 1 (SAG1) from several isolates was characterized by sequence analysis and evaluated for molecular diagnostic identification. AluI restriction digestion of PCR amplicons of the SAG1 gene verified the sequencing results. The putative SAG1 gene from isolates of S. neurona or S. neurona-like parasites varied from 73 to 100% sequence similarity, whereas the SAG1 gene from isolates of Neospora spp. varied from 96 to 98% sequence similarity. PMID- 12581582 TI - Evaluation of a combined immunomagnetic separation/flow cytometry technique for epidemiological investigations of Cryptosporidium in domestic and Australian native animals. AB - A combined immunomagnetic separation (IMS) and flow cytometry (FC) technique was developed for the sensitive detection of Cryptosporidium in faecal samples. The IMS/FC technique was found to be approximately 50-fold more sensitive than formol ether concentration, which is commonly used for Cryptosporidium epidemiological investigations. Of 31 faecal samples from captive animals 16 were found to contain Cryptosporidium oocysts when analysed using the IMS/FC compared to four when using formol-ether concentration (FEC). In a wild population of eastern grey kangaroos Macropus giganteus 66.3% of infected animals were shedding <500oocysts/gfaeces when analysed using IMS/FC. This is below the detection limit for the FEC method. The dispersal of Cryptosporidium in host populations is aggregated, with many individuals shedding low numbers of oocysts and few individuals shedding numbers of oocysts sufficiently high to be detected by FEC. This research demonstrates that the prevalence and oocyst shedding intensity of Cryptosporidium in animal populations will be significantly underestimated using standard detection methods. PMID- 12581583 TI - Inter- and intra-strain variation and PCR detection of the internal transcribed spacer 1 (ITS-1) sequences of Australian isolates of Eimeria species from chickens. AB - The objective of this study was to confirm the presence of seven species of Eimeria involved in chicken coccidiosis in Australia by comparing internal transcribed spacer 1 (ITS-1) sequences, ITS-1 polymerase chain reaction (PCR) methods and to apply phylogenetic analysis to assess evolutionary relationships of Australian isolates. Twenty-two distinct ITS-1 regions of 15 Australian Eimeria isolates were sequenced, and analysed using maximum parsimony, distance and maximum likelihood methods. Poor bootstrap support, resulting from high ITS-1 sequence heterogeneity between all species groups, resulted in polychotomy of the Eimeria species in all three trees generated by these analyses. Percentage identity analyses revealed two distant ITS-1 lineages in both E. mitis and E. maxima at the same levels that separate the two species E. tenella and E. necatrix. One E. maxima lineage consisted of Australian isolates, the other American isolates, with one European sequence (originating from the same isolate) in each lineage. One Australian E. praecox sequence was only distantly related (33% variation) to three E. praecox sequences from Australian and European isolates. Short and long ITS-1 variants were isolated from both E. tenella (cloned line) and E. necatrix isolates with deletions (106 and 73 bp, respectively) in the short variants within the 3' region of the ITS-1 sequence. ITS-1 sequences of strains of both E. brunetti and E. acervulina species varied the least. Apart from E. maxima, all of the ITS-1 sequences of the six remaining individual species clustered to the exclusion of other species in all phylogenetic trees. Published ITS-1 tests for E. necatrix, E. acervulina, E. brunetti and E. tenella, combined with three new tests for E. mitis, E. praecox and Australian E. maxima amplified all respective Australian isolates specifically in a nested format using conserved ITS-1 PCR products as template to improve the sensitivity. All PCR tests were confirmed against a collection of 24 Australian chicken Eimeria isolates and contaminating species were detected in some instances. In conclusion, once the genetic variation between species and strains is determined, the ITS-1 is a good target for the development of species specific assays, but the ITS-1 sequences alone do not seem suitable for the confirmation of phylogenetic inferences for these species. This study reports the first attempt at the analysis of the phylogeny and sequence comparison of the Eimeria species involved in chicken coccidiosis in Australia. PMID- 12581584 TI - Fasciola hepatica: effects on blood constituents and liver minerals in fallow deer (Dama dama). AB - Haematological and serum biochemical constituents as well as the liver mineral concentrations can be affected by Fasciola hepatica. The present study compares the blood and liver constituents in 19 fallow deer with fascioliosis and in 43 without it. Blood samples, gastro-intestinal tract, lungs and liver were taken from 62 male and female fallow deer that were shot, aged 6 months and over, in enclosures, between 1999 and 2001 in Slovenia. Samples were analysed and the means of various parameters of blood and liver were determined. Significant differences in blood were found in the mean cell volume, total serum proteins, sodium, and calcium; and in liver and calcium and copper were higher in animals without fascioliosis. Mean cell haemoglobin concentration, eosinophils, basophils, lymphocytes, monocytes, alanine aminotransferase, urea, glucose and copper in blood were significantly higher in animals with fascioliosis. It has been concluded that some of the values in fallow deer like in other domestic ruminants are the sensitive indicators of liver cell damage in fascioliosis. PMID- 12581585 TI - Preparation of Ehrlichia ruminantium challenge material for quantifiable and reproducible challenge in mice and sheep. AB - The causative agent of heartwater, Ehrlichia ruminantium, is a tick-transmitted pathogen that infects bovine endothelial cells. Due to the obligate intracellular nature of this organism obtaining pure material in sufficient quantities for challenge studies is difficult. A murine model is frequently used to study potential vaccine candidates but giving reproducible challenges in this model for heartwater has always been problematic. We have therefore performed a series of experiments to optimize the parameters governing the reproducibility of challenge material. Two cryoprotectants were compared for the preparation of challenge material, buffered lactose peptone (BLP) and sucrose-potassium-glutamate (SPG). In addition two sources of virulent E. ruminantium were used, infected bovine endothelial cultures and infected mouse spleen homogenates. We also examined practical parameters affecting the reproducibility of challenge experiments: the time it takes to deliver the challenge material, the length of time a mouse remains immune to E. ruminantium challenge, and the effect of a given challenge dose. Finally, we performed a pilot study to determine whether mice could be used to titrate challenge material to be used for experiments in sheep. We found that: (a) E. ruminantium-infected mouse spleen homogenate provides more reproducible challenges than tissue culture material; (b) SPG is a better cryoprotectant than BLP; (c) challenge material should be used within 20min of thawing; (d) it is not essential to use syngeneic material for murine challenge experiments; (e) Balb/c mice are more sensitive to E. ruminantium challenge than C57BL/6J mice; (f) mice immunized by infection and treatment for use as positive immune controls should be challenged within 3 months of immunization; and (g) mice should be challenged with a dose not exceeding 10 LD(50)s. PMID- 12581586 TI - Factors related to cattle infestation level and resistance to acaricides in Boophilus microplus tick populations in New Caledonia. AB - Boophilus microplus, even in the absence of babesiosis, is a major disease of the cattle in New Caledonia where the particular farming system associates continental European breeds and a tropical climate tempered by the Pacific Ocean. In order to have a better understanding of the factors involved in cattle tick infestation, to decrease the possible wastage and use of chemicals and to increase the lifetime of the acaricides, the veterinary authorities investigated the conditions of the chemical treatments. A survey among 148 cattle farms of the whole of New Caledonia was carried out and factors that explain the development of tick resistance and cattle infestation have been determined. From this survey, three typologies for the main characteristics of the farms have been set up, the technical practices of the farmers and the tick control measures applied by the farmers, respectively. Some variables are significantly associated with the tick resistance to deltamethrin but their contribution to the explanation model is always moderate: farms in the south, with a positive resistance gradient from east to west, absence of bush fire and membership to a cattle farmers organization. The more the farmers have intensified their breeding-male castration, weaning, heifer separation, drenching, etc.-and pasture-high stock rate, mowing, extra feeding of the cows, many paddocks, etc.-techniques, the higher was the probability for the ticks in their farm to be resistant to deltamethrin. The technical details of the acaricide treatment had a low contribution to the explanation model. However, the use of a spray generated more resistance than a dip. Furthermore, there is a negative resistance gradient when the farmers increased the treatment interval average. Considering infestation, none of the variables from the three typologies were associated with the two infestation variables (1: semi-engorged tick females and 2: other ticks) at the herd level. However, the seven studied variables-the three typologies, breed, age, body condition score and breeding status-affected significantly the two infestation variables at the cow level, but their predictive ability remained very low (R(2)<3.5%). This result-individual effect more important than herd effect on the infestation-is confirmed by the importance of the variance of the intra-farm factors (99%) when compared with inter-farm factors (1%). Cows of Charolais breed, in poor body condition, old, pregnant or lactating, and those of the farms with irrational and high pressure control of ticks are the most infested. PMID- 12581587 TI - Assay of fipronil efficacy to prevent canine monocytic ehrlichiosis in endemic areas. AB - Our objective was to evaluate the efficacy of fipronil for the prevention of Ehrlichia canis transmission to dogs by Rhipicephalus sanguineus in two endemic areas situated in Africa (Dakar and Djibouti). We carried out controlled trials in kennels for 1 year on 248 dogs, mainly police dogs and military working dogs. Eight groups were studied in a multi-centre study. Fifty five fipronil treated dogs were located in two separated kennels (G3, 37 dogs in Djibouti and G8, 18 dogs in Dakar). G1 (66 dogs) and G2 (60 dogs) were untreated control groups located in Djibouti, whereas G4 (32 dogs), G5 (13 dogs), G6 (18 dogs) and G7 (4 dogs) were the control groups located in Dakar. The epidemiological status of each group is known. G1 and G2 dogs were not kept in kennels, whereas G3, G4, G5, G6, G7, G8 dogs were housed in equivalent kennels. Tick infestation, clinical status and Ehrlichia seroprevalence were assessed during 1 year (duration of the study). Dog treated with fipronil showed neither canine monocytic ehrlichiosis (CME) nor tick infestations. In all groups of untreated control animals, R. sanguineus tick infestations were frequent, particularly in kennels (G5, G6 and G7) as well as morbidity and mortality due to CME. E. canis infection rates were low for fipronil treated animals: 2.7% (1/37) for G3 and 5.5% (1/18) for G8 group. Among control animals, seroprevalence was maximum (100%) in dogs kept in kennels (G5, G6 and G7 groups) and high among native dogs in Djibouti (G1 group): 69.7% (46/66) and in Dakar (G4 group): 50% (16/32). Dogs belonging to expatriate citizens (G2 group) were less likely to be infected: 21.7% (13/60). The comparison of serological results among French army dogs and French citizen dogs that were introduced in Djibouti for an average of 10 months shows a statistically significant (P<0.001) difference. Among fipronil treated animals (G3 group), 2 dogs out of 55 seroconverted (3.6%) compared to 13 out of 60 dogs (21.7%) in the control G2 group. The results of our study indicate the preventative efficacy of a fipronil monthly treatment to avoid CME in endemic areas. Epidemiological data concerning animals that live in the same endemic areas are an example of the serious consequences (in terms of mortality and morbidity) that are related to the absence of efficient methods for tick-control. In order to protect dogs that are in transit in endemic areas against tick transmitted diseases, the use of an adapted acaricide product is recommended. PMID- 12581588 TI - PCR amplification and sequencing of ITS1 rDNA of Culicoides arakawae. AB - The first internal transcribed spacer (ITS1) of nuclear ribosomal DNA from Culicoides arakawae was amplified by PCR, cloned and sequenced. The wDNAsis software was used to analyze the ITS1 sequences of C. arakawae and other nine species of Culicoides, which were obtained from GenBank and EMBL databases. For all species, the lengths of the ITS1 were 316-469 bp, and the G+C contents were 26.79-34.53%. Based on the lengths of the ITS1 sequences, the 10 Culicoides species could be divided into two groups. The first group consisted of C. arakawae, C. albicans, C. cubitalis, C. pulicaris and C. punctatus, and the second group comprised C. impunctatus, C. nubeculosus, C. variipennis, C. grisescens and C. imicola. The lengths for the first group were 316-347 bp and the second group were 457-469 bp. C. arakawae belonged to the first group by its ITS1 sequence length. Sequence analysis revealed that C. arakawae was genetically more similar to the first group than it was to the second group, consistent with results based on sequence length. The alignment of ITS1 (the alignment length was 500 bp including the gaps) sequences showed that there was a highly conserved region, which was between 288 and 388 bp, except for a few insertions and substitutions. These findings have important implications for the molecular identification of C. arakawae, for studying its molecular genetics and epidemiology, and for studying the molecular systematics of Culicoides. PMID- 12581589 TI - Structure of the community of the Strongylidae nematodes in the dorsal colon of Equus caballus from Rio de Janeiro state--Brazil. AB - The structure of the community of Strongylidae nematodes in the dorsal colon of naturally infected horses from the metropolitan region of Rio de Janeiro State was evaluated in 33 samples. Twenty-three species were found: Cylicostephanus longibursatus, Cylicostephanus goldi, Cylicocyclus nassatus and Cyathostomum tetracanthum were the central ones; Cylicostephanus minutus, Cylicostephanus calicatus, Cylicocyclus leptostomus, Cylicodontophorus bicoronatus and Parapoteriostomum euproctus were secondary, and there were 14 satellite species. The community was considered stable, with positive associations and correlations, no significant negative associations and correlations, suggesting little competition among species. PMID- 12581590 TI - Th1 response in BALB/c mice immunized with Dirofilaria immitis soluble antigens: a possible role for Wolbachia? AB - The immune response to filarial infection has been shown to be of both the Th1 and Th2 types. Studies aimed at developing immunization strategies against Dirofilaria immitis infection in dogs have shown that protection against larval challenge is of the Th2 type and that several proteins are recognized by immunized or infected animals. The bacterial endosymbiont Wolbachia, harbored by many filarial species including D. immitis, has recently been shown to interact with the host immune system. Specific antibodies to the Wolbachia recombinant surface protein (WSPr) have been observed in cats infected with D. immitis. In this work the authors have determined cytokine production and antibody response in BALB/c mice inoculated with soluble antigens from third stage larvae or from adult worms of D. immitis. Inoculated mice first produced IFN-gamma followed by a peak in IL-4. Specific antibodies to the Wolbachia protein WSPr were exclusively IgG2a, while antibodies against peptides derived from antigens of D. immitis were in the IgG1 and IgE subclasses. The cytokine response is thus similar to that reported for other filarial infection, where Th1 response shifts towards Th2. Antibody response indicates that Wolbachia may induce preferentially a Th1 response during filarial infection, while nematode antigens may be involved in Th2 response. There is thus an overall agreement with current opinions on the role of bacterial versus nematode molecules in driving the response towards the different directions. PMID- 12581591 TI - Intestinal migrations of Trichostrongylus retortaeformis (Trichostrongylina, Trichostrongylidae) in the rabbit. AB - Observations were made on histological sections of the stomach and small intestine of seven rabbits infected with Trichostrongylus retortaeformis and from one uninfected control rabbit. At 12h post-infection, larvae were found in the small intestine. At first, only a few larvae were observed entering the mucosa through capillaries of the stroma of villi; the majority of larvae remained in the intestinal lumen, within mucus of the crypts. We consider that the presence of the worms in the stroma is the result of a larval migration. From a phyletic point of view, this migration is interpreted as an ancestral memory of the pulmonary migration seen in the primitive Strongylida. PMID- 12581592 TI - The effect of birdsfoot trefoil (Lotus corniculatus) and chicory (Cichorium intybus) on parasite intensities and performance of lambs naturally infected with helminth parasites. AB - Conventionally, farmers rely upon the routine use of anthelmintics to control helminth parasites and their use has proved highly cost-effective. However, several factors, including the emergence of helminths resistant to pharmaceutical anthelmintics, are forcing farmers to seek alternative approaches to parasite control. Studies in New Zealand have shown that some alternative forages may reduce parasitic infestation in sheep. In the current study, it was found that under UK environmental conditions lambs with naturally acquired helminth infections grazing chicory (Cichorium intybus) and birdsfoot trefoil (Lotus corniculatus) had fewer helminth parasites than sheep grazing ryegrass/white clover (Lolium perenne/Trifolium repens). Twelve pure-bred Lleyn male lambs grazed replicated 0.5ha plots of birdsfoot trefoil, chicory or ryegrass/white clover for 5 weeks. Liveweight and faecal egg counts (FECs) were determined weekly and eight lambs per forage were slaughtered at the end of the trial to determine total helminth intensities. Lambs grazing birdsfoot trefoil had a lower FEC on day 7 (P<0.05) and fewer total adult helminths than those grazing the other forages on day 35 (P<0.01). Lambs grazing chicory did not have significantly lower FEC than lambs grazing other forages but these lambs were found to have fewer total adult abomasal helminths than lambs grazing ryegrass/white clover (P<0.001). As the performance of grazing lambs is inversely correlated with the intensity of helminth parasites, these alternative forages could be used to improve the liveweight gain of lambs produced in the UK. Overall, the results support the contention that alternative forages could have a positive role in the control of helminth parasites in sheep, subject to successful agronomic development and integration of these forages into whole farm systems. PMID- 12581593 TI - Ixodes hexagonus is the main candidate as vector of Theileria annae in northwest Spain. AB - Babesia canis and Babesia gibsoni have, until recently, been considered the only piroplasms that parasitise dogs. However, recent reports indicate that "small" Babesia infections in Spanish dogs are surprisingly frequent and molecular phylogenetic analysis indicates that the infecting agent is closely related to Babesia microti. Because the 18SrDNA sequence was not completely identical to that of B. microti, the new name "Theileria annae" was assigned to the canine agent. No information is available regarding the possible vector of the new piroplasm, T. annae. As part of an effort to identify the tick that may transmit T. annae in northwest Spain we asked veterinary surgeons practising in the region to collect and send to our laboratory ticks from dogs visiting their clinics. Seven hundred and twenty ticks collected from dogs of unknown clinical status during 1998 and 636 ticks collected between November 2001 and March 2002 from 38 dogs infected with T. annae and 131 uninfected dogs were identified. Results from the first study indicated that among the Ixodidae, Ixodes hexagonus clearly predominates over Ixodes ricinus (26.11% versus 6.67%). This observation was consistent with results of the second study, in which I. hexagonus was detected in all infected dogs and 71.8% of non-infected dogs and I. ricinus was not detected in either the infected or non-infected dogs. Results from the 2001-2002 study also indicate that the presence of Dermacentor reticulatus adult females is significantly less frequent among infected than non-infected dogs (OR=0.44; 95% CI: 0.21-0.92). On the other hand, I. hexagonus adult females and males are 6.75 and 4.24 times more likely to be detected among infected than non-infected dogs, respectively, with the association being, in both cases, statistically significant (95% CI: 1.97-23.12 and 1.92-9.36, respectively). I. hexagonus emerges as the main candidate as vector of T. annae because it feeds on dogs more frequently than other ticks and because B. microti is transmitted by Ixodes ticks, both in North America and Europe. In the absence of definitive confirmation of this hypothesis, our observations suggest that I. hexagonus might serve the same role as does Ixodes scapularis (=Ixodes dammini), the vector of B. microti in eastern North America. PMID- 12581594 TI - VETSTAT-the Danish system for surveillance of the veterinary use of drugs for production animals. AB - The Danish Ministry of Food, Agriculture and Fisheries funds a monitoring system based on drug usage information collected at the herd level: VETSTAT. VETSTAT is constructed as a relational database and data originates from three sources: pharmacies, veterinarians and feed mills. All administration of drugs for use in animal production is reported on a monthly basis. Pharmacies provided 95% of the total weight antimicrobial compounds used in Denmark in 2001. More than 80% of the antimicrobial compounds reported by pharmacies were sold on prescription to end-users (owners) and included information on animal species, age-group and diagnostic grouping; >90% of the total amount of antimicrobials sold on prescription was used for pigs. In 2001, sales of 96,500 kg of antimicrobials were reported. PMID- 12581595 TI - The need for built-in validation of surveillance data so that changes in diagnostic performance of post-mortem meat inspection can be detected. AB - Time trends in animal-disease surveillance often are evaluated on the basis of crude estimates of apparent prevalence. In addition to possible changes in the true prevalence of the condition, changes in apparent prevalence over time might reflect changes in sensitivity and/or specificity of the diagnostic classification used. To illustrate this, comparative post-mortem meat inspection data from four Danish slaughter plants sampled in 1993-1994 and 1997-1998 were used to obtain latent-class model estimates of the sensitivity and specificity of traditional and extended post-mortem meat inspection of visceral and parietal chronic pleuritis (CP), respectively.True prevalence of CP was estimated for each study period and slaughter plant by latent-class models. Estimated sensitivities of traditional post-mortem meat (TPM) inspection ranged from 28.8 to 61.4% (1993 1994) and 39.2 to 87.3% (1997-1998). An increase in sensitivity with time was seen for all slaughter plants. Estimated sensitivities of extended post-mortem meat (EPM) inspection ranged from 85.7 to 94.8% (1993-1994) and 73.8 to 93.0% (1997-1998). All estimated specificities were >93.3%. The possible association of the estimated true prevalence of CP with time (1993-1994 versus 1997-1998) was investigated with a logistic-regression model with random effects. A slight, but non-significant decrease in the odds of CP from 1994 to 1998 was found (odds ratio=0.9). In this and similar situations, one should consider conducting ongoing double-classification of samples of units followed by statistical estimation of true prevalences, sensitivities and specificities, so that decisions can be based on such estimates rather than on crude apparent prevalences. PMID- 12581596 TI - Factor analysis of minimum-inhibitory concentrations for Escherichia coli isolated from feedlot cattle to model relationships among antimicrobial resistance outcomes. AB - Factor analysis was used to assess relationships in the minimum-inhibitory concentration among 17 antimicrobials tested on isolates of Escherichia coli isolated from 360 faecal samples obtained from feedlot cattle. Six factors were extracted using maximum-likelihood factor analysis. The factors were interpretable antimicrobial groupings based on class of antimicrobial and previously described associations. New-generation cephalosporins, older generation beta-lactams, fluoroquinolones and aminoglycosides grouped separately as classes of antimicrobials on four of the six factors. One of the remaining factors was a grouping of antimicrobials that had been identified as being related in previous feedlot studies. The last factor was a grouping of three of the five antimicrobials that comprise the antimicrobials found in penta-resistant strains of Salmonella Typhimurium. The factor analysis described patterns in the MIC data that would not have been apparent if only antimicrobial-resistance data categorized as susceptible-resistance had been analysed. PMID- 12581598 TI - The foot-and-mouth disease epidemic in The Netherlands in 2001. AB - An outbreak of foot-and-mouth disease (FMD) in Great Britain was reported on 21 February 2001, followed by an outbreak of FMD in The Netherlands a month later. This Dutch index outbreak occurred on a mixed, veal-calf/dairy-goat farm in Oene, in the central part of The Netherlands. The most-likely route of infection was the import of Irish veal-calves to this Dutch herd via an FMD-contaminated staging point in France. With hindsight, more herds seemed to be infected by the time the index outbreak was confirmed. The regular EU control measures were implemented, in combination with pre-emptive culling of herds within 1km of each outbreak. Nevertheless, more outbreaks of FMD occurred. Most of the virus infections on those farms were "neighborhood infections". Because the situation seemed out of control locally and the destruction capacity became insufficient, it was decided to implement an emergency vaccination strategy for all biungulates in a large area around Oene to stop further spread of the virus. All susceptible animals on approximately 1800 farms in this area were vaccinated. All farms subsequently were depopulated, starting from 2 weeks after vaccination. In total, 26 outbreaks were detected (the last outbreak on 22 April 2001). In total, approximately 260,000 animals were killed. PMID- 12581597 TI - Validation of a Neospora caninum iscom ELISA without a gold standard. AB - Neospora caninum is an intracellular parasite which causes abortion in cattle worldwide. One problem in the validation of the different methods for demonstration of this parasite is the lack of an appropriate gold standard. To validate an immunostimulating complex (iscom) enzyme-linked immunoassay (ELISA) used to detect antibodies to N. caninum, sera from 244 cattle in five Swedish dairy herds infected with N. caninum were analysed. The sera also were analysed by a standard indirect-fluorescent antibody test (IFAT). The results obtained by the two tests were compared using the Gibbs sampler. Gibbs sampling is a latent class approach based on Bayesian statistics; neither test is assumed to be more correct in stating the true status of infection. The Gibbs sampler was run using both informative and non-informative prior probabilities. We also simulated different cut-offs in the iscom ELISA (providing data to inform selection of optimal cut-off values for different applications). The ELISA produced fewest incorrect test results over all at a cut-off value of 0.200. The sensitivity and specificity at this cut-off were 99 and 96%, respectively. The IFAT had a high specificity (99%) but a lower sensitivity (78%) than expected-confirming that the IFAT cannot be treated as a true gold standard. Sensitivity and specificity of the ELISA were presented in a two-graph receiver operating characteristic (TG ROC) plot. Any cut-off between 0.150 and 0.300 will have both sensitivity and specificity > or =95%. Optical densities of < or =0.150 and > or =0.550 (or > or =0.350) were suggested as limits to rule out and rule in infection, respectively. PMID- 12581599 TI - Prevalence of bovine tuberculosis in cattle in different farming systems in the eastern zone of Tanzania. AB - The prevalence of bovine tuberculosis (BTB) infection in cattle was investigated in pastoral and intensive production systems in the eastern zone of Tanzania. The prevalence of BTB infection as determined by single comparative intradermal tuberculin test (SCITT) was 1.3%, whereas the non-specific infection prevalence was 6%. In the pastoral sector, the prevalences were 1 and 7%; under intensive systems, they were 2 and 6% for BTB and non-specific infections, respectively. The prevalences were significantly higher in the intensive than pastoral production systems. However, the prevalence of BTB infection was higher in the small-scale (3%) than in other production systems (0.6-1.1%). Non-specific infections were lowest in the small-scale dairy sector (4%) than in other dairy production systems (6-11%). Out of 143 herds tested, 10 and 28% of herds in the intensive systems (small, medium and large farms) and 17 and 88% of the herds in the extensive system (pastoral farms) had both BTB and non-specific infections, respectively. PMID- 12581600 TI - Aqueous humor dynamics and the iris. AB - Solutes concentration in the aqueous humor of the eye is highest near the ciliary processes in the posterior chamber (p.c.) and lowest near the trabecular meshwork in the anterior chamber (a.c.). The high osmotic gradient across the semipermeable iris causes it to easily move forward at this location and occasionally occlude the iridocorneal angle. Vector analysis of the forces generated by miosis helps explain on osmotic grounds why it reduces the intraocular pressure, while at the same time the analysis elucidates the reason for the higher frequency of angle closure glaucoma when the anterior chamber is shallow. PMID- 12581601 TI - An effective veterinary model may offer therapeutic promise for human conditions: roles of cortisol and thyroid hormones. AB - For nearly three decades, the author has treated multiple serious diseases of cats and dogs by correcting an unrecognized endocrine-immune imbalance originating with a deficiency or defect of cortisol. The cortisol abnormality creates a domino effect on feedback loops involving the hypothalamus-pituitary adrenal axis. In this scenario estrogen becomes elevated, thyroid hormone becomes bound, and B and T cells become deregulated. Diseases with this aberration as a primary etiological component range from allergies and strange behavior to severe cases of autoimmunity and cancer. Successful treatment and control, even in critical cases, have been consistently achieved with a long-term physiologic (not pharmacologic) replacement with cortisone along with thyroid hormone (in dogs). The treatment represents a major healing modality for many seemingly unrelated chronic diseases of animals. In humans, this endocrine-immune dysfunction appears to exist and, as in veterinary medicine, has been overlooked by researchers and clinicians. Testing and treatment patterned after the animal model may offer significant clinical benefits for challenging human afflictions. PMID- 12581602 TI - Failure of blood-thymus barrier as a mechanism of tumor and trophoblast escape. AB - A major process through which the immune system becomes tolerant to self-proteins involves the deletion of self-reactive clones in the thymus, but clonal deletion is not single mechanisms of thymic tolerance. There is now much evidence that intrathymic antigen expression results in anergy induction of T helper type-1 (Th1) clones in the periphery. Blood-thymus barrier is most important structure for prevention of unwanted penetration of antigens into the thymus. Impermeability of the barrier restrain induction of acquired thymic tolerance on unwanted antigens like microbes and tumor cells. Nevertheless, one of most important mechanism of tumor and trophoblast escape is in anergy of Th1 cells and in Th2 cells domination. Many mechanisms are included in disarrangement of Th1/Th2 balance in pregnancy and tumor bearers, but one of possibility is in failure of blood-thymus barrier. Possible consequences of blood-thymus barrier failure are trophoblast-specific or tumor-specific antigens penetrate into the thymus, deletion or anergy of antigen-specific clones and acquired thymic tolerance induction. Blood-thymus barrier is variable structure in anatomical and functional sense so that in certain condition foreign antigens probably can permeate across the barrier. Probability that some factors like hormones, cytokines, prostaglandine and neuromediators can affect blood-thymus barrier permeability and contribute in mechanisms of trophoblast and tumor escape is real but relatively unexplored. PMID- 12581603 TI - Consanguinity as risk factor for cervical carcinoma. AB - Cervical carcinoma is caused by human papillomavirus (HPV). Among the risk factors for HPV infection are having multiple sex partners and sex partners who themselves had multiple sex partners. Women married to relatives are more likely to become infected with HPV and develop cervical carcinoma. Consanguineous spouses have a mild sexual aversion to each other that leads to sex avoidance and increases likelihood of satisfying sexual desire outside the marriage. Sexual aversion develops as a result of spouses growing together in early childhood, which triggers biological imprinting of Westermarck. Westermarck's effect prevents incest in nuclear family. In consanguineous family, it extends to spouses, producing mild sexual antipathy. Because of high prevalence of consanguinity worldwide, it is important to test this hypothesis. PMID- 12581604 TI - The effect of isoprenoid side chain length of ubiquinone on life span. AB - The isoprenoid side chain length of ubiquinone (Q) can have an effect on the life span of mammals. The short living mouse and rat have Q(9), while primates have Q(10) as the major form. Ubiquinones (Qs) having longer hydrophobic tail are likely more imbedded in the mitochondrial inner membrane than the ones having shorter tails. In case of short tail length, ubisemiquinone (Q(*-)) produced during electron transport can be more exposed to the aqueous phase on both sides of the membrane, generate more superoxide radical and damage the neighbouring macromolecules. Considering the inefficient subcellular distribution of exogenous Q, production of transgenic animals synthesizing Qs having longer than 10 isoprenoid units (Q(>10)) can increase their life span. PMID- 12581605 TI - Stress-related preeclampsia: an evolutionary maladaptation in exaggerated stress during pregnancy? AB - The authors hypothesize that preeclampsia is a stress-related disease and an evolutionary maladaptation of exaggerated stress during human pregnancy. Epidemiologic studies show that relative risk for preeclampsia is increased in many stressful situations. Many risk factors for preeclampsia are stress-related. Low-stress situations, on the contrary, are protective. Stress in pregnancy corroborates all physiopathologic theories for preeclampsia; it does not contradict them. Animals exposed to intense stress show many characteristics of preeclampsia, and some animal models for human preeclampsia have been proposed. The stress-alarm reaction is protective for survival in animals. But the evolutionary maladaptation of this intense stress could lead to preeclampsia in humans. PMID- 12581606 TI - Hypoxia-induced upregulation of the glycolytic enzyme glucose-6-phosphate isomerase perpetuates rheumatoid arthritis. AB - Intra-articular hypoxia in the inflamed rheumatoid joint is associated with increased cell proliferation, enhanced metabolism and compromised vascular perfusion. Recent clinical studies using direct measurements of hypoxia in rheumatoid joints have delineated up to 20% of soft tissue pO(2) readings as below 10mm Hg. Increased markers for glycolysis exist in rheumatoid synovial fluid and upregulation of tissue glycolytic enzymes occurs in a rat model of synovitis. Recent reports show arthritis is provoked by linked T and B cell lymphocyte recognition of the glycolytic enzyme glucose-6-phosphate isomerase (GPI). This suggests an unusual physiological feature of rheumatoid joints leads to autoimmune destruction. In this report I suggest that hypoxia, within the rheumatoid joint, leads to upregulation of the glycolytic enzyme GPI which in turn perpetuates rheumatoid arthritis. PMID- 12581607 TI - Minor thalassemia may be a risk factor for impulsiveness. AB - Frequent studies have demonstrated the relationship between low cholesterol levels and increased aggression as well as suicide. In primates, cholesterol reduction has been linked to increased behavioral acts of aggression. Patients with low cholesterol show hypersensitivity in the detection of anger and sadness. It is documented that minor thalassemics have lower serum cholesterol levels than the general population. Therefore, it could be postulated that minor thalassemics may be predisposed to impulsiveness due to the decrement of serum cholesterol levels. PMID- 12581608 TI - Does infection with hepatitis A virus provide protection against hepatitis E virus? AB - Hepatitis A virus (HAV) and hepatitis E virus (HEV) share some common epidemiologic features, but the dramatic age-related difference in their disease prevalences suggests that the immune response to HAV may provide some cross protection against HEV. PMID- 12581609 TI - Finger and toe ratios in humans and mice: implications for the aetiology of diseases influenced by HOX genes. AB - The differentiation of gonads, fingers, and toes is influenced by HOXA and HOXD genes. Therefore variation in the development of the gonads, and their fetal products such as testosterone, may be reflected in the morphology of the fingers and toes. One trait, the relative length of the second and fourth digits (2D:4D), shows sex differences (lower values in males) which are determined early, and 2D:4D has been found to correlate with fetal growth, sperm counts, family size, autism, myocardial infarction, and breast cancer. HOX genes are highly conserved in mammals and they influence the differentiation of all the fingers and toes. We suggest that (a) 2D:4D and other ratios of finger and toe length show sex differences throughout the mammals including humans and mice, (b) finger and toe ratios correlate with sex determination, the fetal production of sex steroids, and fetal programming of disease, and (c) HOX gene influences on sex determination, the morphogenesis of the urinogenital system, fertility, haematopoiesis, and breast cancer suggests that finger and toe ratios in humans and mice may correlate with many sex dependent diseases. PMID- 12581610 TI - Etiopathogenesis of pediatric Crohn's disease. Biologic pathways based on interactions between genetic and environmental factors. AB - The incidence and prevalence of Crohn's disease (CD) among children in developed countries is extremely high. Although, dietary factors have long been implicated, to date no single causative element has been identified. Risks or benefits for CD from dietary elements would depend on their appropriate metabolism within the gut by xenobiotic metabolising enzymes (XME). The metabolising capacity among individuals differs and is determined by the level and/or activity of the specific XME. As the latter is the function of the DNA variants that code them, we propose that certain individuals exposed to dietary elements are likely to be more or less susceptible for CD. We highlight this hypothesis by describing gene environment interaction (GxE) mechanisms that could determine susceptibility for CD. We suggest that investigating these mechanisms will be paramount for the appropriate identification of susceptible populations, such that preventive and/or therapeutic interventions could be adequately targeted. PMID- 12581611 TI - Diet quality and risk of neural tube defects. AB - Many studies have examined the impact of single nutrients on neural tube defect (NTD) risk, particularly folate. The impact of dietary patterns or nutrients in combination has received much less attention. This study examines the association of diet quality with NTD risk, using food frequency data from a population-based case-control study of NTDs (n=454 cases and 462 controls). The diet quality score was based on low (<10th percentile among controls) versus high (>90th percentile) values for intakes of iron, vitamins B(6) and A, calcium, folate, and percentage of kilocalories from fat and from sweets (range=0-14). Women with a low score (i.e., <4, or <10th percentile) had an elevated risk of an NTD-affected pregnancy (odds ratio 1.6, 95% CI 1.0-2.6). Stratified analyses suggested that the effect may be restricted to certain groups of women who may be at greater nutritional risk (i.e., women who did not take vitamin supplements or regularly consume breakfast cereals before pregnancy). PMID- 12581612 TI - Can stress provide protection to pancreatic beta-cells and prevent diabetes? AB - A wide variety of stresses are prevalent in the environment that could change the course and phenotypic expression of metabolic diseases. Amongst the various stresses the oxidative stress plays a pivotal role in the disease progression through free radical generation and may lead to various metabolic disorders such as non-insulin dependent diabetes mellitus, cardiovascular diseases, hypertension obesity, stroke, etc. Psychological stress has been implicated as a root cause of several psychosomatic disorders. Thus there cannot be a life without stress. Under such a scenario of stressful conditions further fueled by life style changes we propose to counteract any kind of stress by another milder form of stress that is likely to protect the cells from drastic effects of severe stress. Here we hypothesize that the beta-cells of islets of Langerhans can be protected from the diabetogenic insults and oxidative stress by inducing a protective stress response such as heat shock through dietary interventions. PMID- 12581613 TI - Immunoengineering: a credible mechanism for CAMPATH-1H action in bone marrow and organ transplantation and the implications for treatment of the immune dysfunction AIDS. AB - Immunoengineering is a term coined to represent the mostly future ability to use or target the immune system's natural components, with emphasis on the regulatory components, to up or down regulate the immune system's attack against specific proteins associated with an unwanted pathology or immune occurrence. It will constitute manipulating parts of the immune system, mostly those specific for the disease associated antigen(s) and generally of a regulatory nature, in various immunological locale or the whole body compartment, to achieve a disease free state for the patient. The number of practical applications awaiting the mastery of immune components as regulatory therapeutics is enormous and immunoengineering should provide treatments in a wide range of disease categories. HIV is a disease where this discipline could provide a quick cure, even eradication of the virus. A potential cheap solution to HIV infection, based on using immunoengineering and adaptable to the infrastructure problems of the Third World is highlighted in the following because of the health emergency that exists in the Third World. PMID- 12581614 TI - An editorial for the HIV infected: immune regulation. The implications for treatment of HIV infection and the potential role of T cell suppressor pathways instead of apopstasis, anergy or direct CD4 T cell deletion in AIDS pathogenesis. AB - The AIDS crises makes for an interesting study of the interactions of activists, researchers, caregivers, the press, politicians and other groups in society. In the popular press and elsewhere there has been a growing movement since the inception of the disease to place efforts against AIDS as the highest standard of our benevolence, proficiency and determination when confronting a disease. Because of the potential benefit in understanding the interactions of groups and how research progressed and failed during the AIDS crises, differing perspectives should be recorded on this matter. The person currently in most need of an objective viewpoint on AIDS research, including an outlook on the past, as well as the present and future, is the person who is infected with HIV. While many of the AIDS related bureaucracies, scientists and celebrities have achieved consecration in the fight against HIV, a question arises whether this praise is deserved and really due to their keeping the AIDS patient as the first matter of the heart and mind; or whether they have merely achieved what bureaucracies and celebrity are best at, making society and those they should serve, think that they cannot do with out them.The following editorial chronicles a scandalous intellectual failure of immunologists in the fight against HIV. It delineates potential areas of concern for the HIV infected patient in the present and future, which may be important directions in the fight against HIV, both for treatment to evolve to the ideal and for an economically viable treatment for the Third World. PMID- 12581615 TI - The neurochemical hypothesis of 'theory of mind'. AB - This paper aims to explore the neurochemical basis of the ability to represent one's own or other's mental states such as intentions, beliefs, wants and knowledge, an ability often referred to as 'theory of mind'. Based on neurochemical and psychopharmacological investigations in autism and schizophrenia, pathologies in which this ability is impaired, it is hypothesized that 'theory of mind' abilities are contingent on the integrity of the serotonergic and dopaminergic system. This hypothesis is discussed in light of the system's neurochemical properties and role in cognition. It is suggested that specific abnormalities to this system can account for differences in the profile of 'theory of mind' impairments that may exist among patients belonging to different pathologies. PMID- 12581616 TI - Nitroxyl radicals and malignant pigment melanoma. AB - Our studies showed that the spin-labeled (SL) compounds (free stable nitroxyl radical derivatives) accumulate predominantly in pigment melanoma and some of them possess high antimelanoma activity (100% curability of tumor bearing animals) and lower general toxicity. Taking into account our results we describe a proposed mechanism of the spin-labeled accumulation in melanoma, their antimelanoma activity and lower toxicity. PMID- 12581617 TI - Can we identify novel angiogenesis inhibitors from cultured embryonic cells? AB - The first vascular structures in embryos are formed by vasculogensis, the de novo formation of blood vessels from angioblasts. However, angiogenesis, the sprouting of capillaries from pre-existing blood vessels, are also involved in the development of embryonic vascular system in certain organs, such as the kidney or the brain. These facts led us to speculate that embryonic tissues could produce potent angiogenesis inhibitors as well to refine the primitive vascular development. In the present study, we found that two different types of cultured human embryonic cells, 293T and WI-38, produced soluble factors that completely inhibited angiogenic effects of vascular endothelial growth factor on microvascular endothelial cells. Since angiogenesis is related to various pathological states, including tumor growth and metastasis and diabetic retinopathy, to identify novel potent angiogenesis inhibitors from cultured embryonic cells by differential display techniques or DNA microarray technology might be a valuable strategy to develop a novel therapeutic approach. PMID- 12581618 TI - 7-oxo-DHEA and Raynaud's phenomenon. AB - Patients with Raynaud's phenomenon have abnormal digital vasoconstriction in response to cold. The pathogenesis remains unknown but may involve a local neurovascular defect leading to vasoconstriction. Diagnosis of primary Raynaud's phenomenon is based on typical symptomatology coupled with normal physical examination, normal laboratory studies and lack of observable pathology by nail fold capillaroscopy. Secondary Raynaud's phenomenon is known to occur associated with several connective tissue diseases, vascular injury due to repeated vibrational trauma, and other causes which produce demonstrable vascular and microcirculatory damage. Treatment of Raynaud's symptoms is conservative and aimed at prevention of attacks. Patients are advised to remain warm and, if possible, to live in warm climates. We suggest that an ergogenic (thermogenic) steroid, 7-oxo-DHEA (3-acetoxyandrost-5-ene-7,17-dione), which is available without prescription as the trademarked 7-keto DHEA, may be very helpful in prevention of primary Raynaud's attacks by increasing the basal metabolic rate and inhibiting vasospasm. PMID- 12581619 TI - Diaphragm pacing during prolonged mechanical ventilation of the lungs could prevent from respiratory muscle fatigue. AB - Respiratory muscle fatigue after prolonged mechanical ventilation of the lungs is a common occurrence. This is probably due to disuse atrophy. The standard technique used to overcome this problem is to gradually wean the patient from the respirator over the course of several days or weeks. HYPOTHESIS: based on experience with the rehabilitation of skeletal muscles during long-term immobilisation of the extremities following trauma or surgery and on experience with diaphragm pacing in quadriplegics, intermittent electrical or magenetic stimulation of the diaphragm while the lungs are still being mechanically ventilated may be beneficial. The result of such stimulation would be to preserve the dysfunctioning diaphragm from atrophy. CONSEQUENCES: dependence on the respirator, length of intubation time, risk of complications and length of stay in the ICU would be considerably reduced. PMID- 12581620 TI - Hypothesis that vagal reinervation of diaphragm could sensitise it to electrical stimulation. AB - The hypothesis proposed is that restoration of functional capacity of denervated diaphragm may be achieved by reinervating it with vagus nerve. Following trauma, carcinomatose infiltration, and/or large thoracic surgery and neck surgery, phrenic nerve is frequently injured. Reinervation even in the most favourable conditions would not follow and diaphragm would rest permanently denervated and paralysed. This results in unilateral or bilateral paralysis of diaphragm. In principle, intermittent electrical stimulation of the phrenic nerve or diaphragm could elicit regular diaphragm contractions and maintain satisfactory respiration. While this technique could be used in upper motor neurone injury, in lower motor neurone injury and denervated diaphragm, that imposes too high electrical resistance, direct diaphragm pacing is practically impossible. In these cases, long term artificial ventilation is often necessary. Nevertheless, those patients are at high risk to suffer from atelectasis and respiratory infections. We project a hypothesis that reinervation of denervated diaphragm by vagus nerve could re-establishes its sensitivity to intramuscular electrical stimulation and may allow stimulation of the diaphragm by implanted pace-maker electrodes. An appropriate electrical stimulation might then be possible and diaphragm pacing could replace prolonged artificial ventilation in those patients. Restoration of functional capacity of denervated diaphragm could open a perspective for long term diaphragm pacing in patients with irreversible phrenic nerve injury and diaphragm paralysis. PMID- 12581621 TI - Neuro-endocrine neoplasms as a result of gene desuppression rather than of a specific cell type of origin. AB - Neuroendocrine differentiation more closely approximates a process of acquisition of features that ambiguously recapitulate certain cellular biologic properties that transgress normal lines of differentiation as these are generally recognized. In fact, it might be more realistic to consider neuroendocrine tumors the result of a process of progressive desuppression via pathways of altered gene expression that would perhaps reflect basic and dynamic aspects of neoplastic transformation and progression in general. In a final analysis, the character and degree of differentiation of a neoplasm might actually often constitute an anomalous combination of features outside the range of normal differentiation pathways and more an expression of an assorted mixture of partially suppressed or desuppressed gene expression profiles as these affect neoplastic cell pathobiology. PMID- 12581622 TI - Are tau and alpha synuclein filamentous inclusions in neurons and glia analogous to accumulation of myosin/actin filaments in myofiber hypertrophy? AB - Degenerative disruption of neuronal networks would appear to constitute a pathogenetic series of mechanisms that would adequately account for the effects of neurodegeneration in its full complexity clinically and pathologically beyond any consideration of the neuron as the basic unit of involvement. In this sense, neurodegeneration as reflected in terms of neurofibrillary tangles and Lewy bodies might actually be a process of integral tau opathy and synucleinopathy that in probably multiple different ways in the individual patient contributes to a disruption of neuronal interactions as an integral neuronal network. In terms therefore of aggregation of neurofilaments or of synuclein filaments it might be valid to consider simple biophysical phenomena of inclusion body formation in neurons directly analogous to myosin/actin accumulation in myofibers in an overall common system of neuronal and myofiber biology and pathobiology. PMID- 12581623 TI - The halting arrival of left-handed Z-DNA. AB - Forty-nine years ago Watson and Crick proposed a double-stranded (ds-) model for DNA. This double helix has become an icon of molecular biology. Twenty-six years later, Rich accidently discovered Z-DNA, an exotic left-handed nucleic acid. For many years thereafter, this left-handed DNA was thought to be an artifact. DNA is no longer looked upon as a static molecule but rather an extremely dynamic structure in which different conformations are in equilibrium with each other. Many researchers have spent the last two decades characterizing this novel left handed DNA structure. Now many investigators are beginning to accept the possibility that this novel ds-DNA conformation may play a significant in vivo role within eukaryotic and prokaryotic cells. However, more research needs to be performed before it is absolutely accepted by all in the scientific community. PMID- 12581624 TI - The significance of the nitric oxide in electro-convulsive therapy: a proposed neurophysiological mechanism. AB - Electroconvulsive therapy (ECT) is used to treat patients with major depressive disorder, manic episodes and other serious mental disorders. Virtually every neurotransmitter system is affected in ECT. The significance of the nitric oxide (NO), which has an established role as a neurotransmitter, neuromodulator, and an intraneuronal second messenger, in ECT is still not clear. We described the involvement of NO in long-term potentiation, the N-methyl-D-aspartic acid (NMDA) receptor activity, regulation of cerebral blood flow, and the hypothalamic pituitary axis and propose that this involvement is critical in ECT's efficiency, treatment refractoriness, and neuropsychological sequelae by its influences on these systems. Nitric oxide's significant role in other pathophysiological mechanisms has led to current therapeutic protocols and may be applicable in this setting. PMID- 12581625 TI - Inertia of endocrine systems due to hormone binding to circulatory proteins. AB - It is often presumed that the main role of hormone binding to albumins and binding proteins (BPs) is to reduce oscillating levels of free hormone molecules and to transport steroid hormones. This paper is an attempt to define possible consequences of hormone molecules binding to carrier proteins in circulation. Binding to albumins and BPs prevents exact and quick control of hormone actions. Hormones without significant protein binding govern vital and fast acting regulatory mechanisms (blood glucose or calcium) in which any added inertia might be dangerous. In the presented model, the added inertia for a partially bound hormone (H) is defined as: H(bound)/H(free). Values, calculated from the reported data, range from 0.4 for GH to more than 2000 for T(4). In comparison to albumins, high-affinity BPs make more stable reserve that would cover periods of low or no hormone secretion. At the same time, hormone molecules are taken away from the blood level control and thus might be considered sequestrated. For hormones without protein binding, the well-perfused areas of the body, or the areas with increased capillary permeability, would be more exposed, making an uneven distribution among target tissues. For the hormone that binds blood proteins, places of secretion and tissue perfusion become unimportant, since the hormone is being liberated anywhere in the circulation (i.e., for strongly bound IGFs, IGF binding proteins do not just stabilize proinsulin actions of IGF-1, but also make all parts of body to be under the same exposure to liberated IGFs, an important feature to promote a symmetrical bone growth). Estrogens are known to stimulate liver secretion of different BPs. A possible explanation is that in the follicular phase there is a small initial mass of granulosa cells, and it takes time to saturate free estrogen carriers, before the normal free hormone level can be reached and FSH secretion inhibited. Less inert peptide inhibin might suppress FSH before free estrogens reach the required level. Without inhibin suppression, an increased FSH level with an increased number of growing follicles can be expected. Estrogens increased production of BPs augments inertia of the estrogen loop and possibly modulates the FSH/estrogen negative feedback. PMID- 12581626 TI - Human stem cells as targets for the aging and diseases of aging processes. AB - While many theories have been proposed for the aging process, and many debates on the matter of aging and the diseases of aging being either the result of the same or independent processes, most have not considered humans as a hierarchical system made up of cybernetically interacting levels of organization. To understand the aging process and the diseases of aging, one must view the human as the result of the total genomic DNA in the single fertilized egg that proliferates, differentiates and develops into an individual of about 100 trillion cells, organized by different cell types (pluri-potent stem cells, progenitor stem cells, terminally differentiated cells) into multiple tissue, organ and organ systems which interact with each other via endogenous factors and with exogenous factors. Our hypothesis is that both aging and diseases of aging are dependent of the normal functioning of the pluri-potent stem cell pool. Specifically, the concept involves the cybernetic feedback between the 'quantity' of the stem cell pool in each tissue niche with the 'quality' of the stem cells in the pool. The process of gap junctional inter-cellular communication (GJIC), which has been implicated in the evolution from the single cell organism to the multi-cellular organisms, requiring growth control, differentiation, apoptosis, adaptive response capability of differentiated cells and senescence, is speculated to be a shared mechanism in stem cell biology and in many chronic disease processes (teratogenesis; carcinogenesis, atherogenesis, diabetigenesis, etc.). Specifically, stem cells are assumed to be 'immortal' until induced to express their connexin genes and have functional GJIC, at which time they can differentiate and become 'mortal'. As long as the stem cells are communicating with their differentiated daughters via some extra-cellular soluble negative growth factor, the homeostatic control of their growth and differentiation is maintained for the organism. However, if the stem cell pool is depleted by any process, replacement of tissue due to wear and tear is diminished. The dependence of this tissue/organ to maintain homeostatic control of other organ systems then diminishes, leading to 'systems failure'. In addition, if the stem cells in the pool have been exposed to agents that prevent the normal terminal differentiation of that cell, but whereby these 'initiated' stem cells can be expanded in any tissue, clones of partially differentiated and non-functional appear in the tissue. This diminishes the efficacy of that tissue to function properly and, thereby, also contributes to 'system failure' by contributing to the breakdown of homeostatic organ system control. One clear example, that of carcinogenesis, illustrates this point. PMID- 12581627 TI - Role of androgens in mild cognitive impairment and possible interventions during andropause. AB - Mild cognitive impairment (MCI) is becoming fashionable as a diagnosis, representing a state of cognitive decline associated with negligible functional loss. MCI is important as it often precedes Alzheimer's disease (AD). Recognizing MCI may lead to preventive strategies that can delay the onset of AD. Many patients who transition into andropause report problems with their memory. There is strong evidence from basic sciences and epidemiological studies that both estrogens and androgens play a protective role in neurodegeneration. The evidence from small prospective clinical trials lends support to the role of hormones in improving cognitive function. The improvement in cognitive function with hormones is subtle and often not measurable on standard neuropsychological batteries. Patients have reported memory improvements in both declarative and procedural domains after being on hormonal replacement. Functional changes and vascular changes can be detected after hormonal replacement with more sophisticated imaging of the brain like positron emission tomography (PET) scans. We hypothesize androgens and perhaps selective androgen receptor modulators as future treatment options for MCI in aging males. PMID- 12581629 TI - Insertion of hydrophobic membrane proteins into the inner mitochondrial membrane- a guided tour. AB - Only a few mitochondrial proteins are encoded by the organellar genome. The majority of mitochondrial proteins are nuclear encoded and thus have to be transported into the organelle from the cytosol. Within the mitochondrion proteins have to be sorted into one of the four sub-compartments: the outer or inner membranes, the intermembrane space or the matrix. These processes are mediated by complex protein machineries within the different compartments that act alone or in concert with each other. The translocation machinery of the outer membrane is formed by a multi-subunit protein complex (TOM complex), that is built up by signal receptors and the general import pore (GIP). The inner membrane houses two multi-subunit protein complexes that each handles special subsets of mitochondrial proteins on their way to their final destination. According to their primary function these two complexes have been termed the pre sequence translocase (or TIM23 complex) and the protein insertion complex (or TIM22 complex). The identification of components of these complexes and the analysis of the molecular mechanisms underlying their function are currently an exciting and fast developing field of molecular cell biology. PMID- 12581628 TI - Mathematical analysis of a pharmacodynamic model without plasma concentrations to extend its applicability. AB - Using a mathematical approach, we analyzed the behavior of the PD model originally described by Bragg et al. The effect was dose-dependent modified until a maximum value (E(max)) was reached. Further increments in dose prolonged the E(max), but the recovery phase did not increase beyond a calculable asymptope. In the absence of plasma concentrations, it was impossible to distinguish the rate of plasma equilibration with the effect compartment (k(e0)) from the rate of drug elimination (k(e1)). Variations on the sigmoidicity affected both the onset and offset of drug effect. Sigmoidicity and the slowest rate constant had identical effects on the spontaneous reversion of the effect, as judged by the recovery index. The IR(50), the index of potency, merely shifted the dose-response relationship to the left or right. Changes in IR(50) were compensated for by making the same proportional changes in dose. PMID- 12581630 TI - Some motile properties of fast characean myosin. AB - We improved a motility assay system by using an affinity-purified antibody against the C-terminal globular domain of characean myosin. This improvement allowed us to study the sensitivity to ionic strength or the processivity of characean myosin. The sliding velocity of actin filaments on a characean myosin coated surface was unaffected by ionic strength. This property is unlike that of skeletal or smooth muscle myosin and suggests that the binding manner of characean myosin to actin is different from that in other muscle myosins. The sliding velocity decreased when the MgADP concentration was raised. The extent of inhibition by MgADP on the motile activity of characean myosin was almost the same as in skeletal muscle or cardiac myosin. The number of sliding filaments on the characean myosin-coated surface decreased drastically with a decrease in the motor density. The motor density required to produce a successful movement of actin filament was about 200 molecules/microm(2). These results suggest that the characean myosin is not a processive motor protein. PMID- 12581632 TI - Interaction of T4 AsiA with its target sites in the RNA polymerase sigma70 subunit leads to distinct and opposite effects on transcription. AB - Bacteriophage T4 AsiA is a homodimeric protein that orchestrates a switch from the host and early viral transcription to middle viral transcription by binding to the sigma(70) subunit of Escherichia coli RNA polymerase holoenzyme (Esigma(70)) and preventing promoter complex formation on most E.coli and early T4 promoters. In addition, Esigma(70)AsiA, but not Esigma(70), is a substrate of transcription activation by T4-encoded DNA-binding protein MotA, a co-activator of transcription from middle viral promoters. The molecular determinants of sigma(70)-AsiA interaction necessary for transcription inhibition reside in the sigma(70) conserved region 4.2, which recognizes the -35 promoter consensus element. The molecular determinants of sigma(70)-AsiA interaction necessary for MotA-dependent transcription activation have not been identified. Here, we show that in the absence of sigma(70) region 4.2, AsiA interacts with sigma(70) conserved region 4.1 and activates transcription in a MotA-independent manner. Further, we show that the AsiA dimer must dissociate to interact with either region 4.2 or region 4.1 of sigma(70). We propose that MotA may co-activate transcription by restricting AsiA binding to sigma(70) region 4.1. PMID- 12581633 TI - Destabilization of the HIV-1 complementary sequence of TAR by the nucleocapsid protein through activation of conformational fluctuations. AB - The nucleocapsid protein NCp7 of HIV-1 possesses nucleic acid chaperone properties that are critical for the two obligatory strand transfer reactions required for the synthesis of a complete proviral DNA by reverse transcriptase. The first DNA strand transfer relies on the destabilization by NCp7 of double stranded segments of the transactivation response region (TAR) sequence at the 3' end of the genomic RNA and the complementary sequence cTAR at the 3' terminus of minus strong-stop DNA, the early product of reverse transcription. In order to determine the dynamics of NCp7-mediated nucleic acid destabilization, we investigated by time-resolved fluorescence spectroscopy and two photon fluorescence correlation spectroscopy, the interaction of a doubly labeled cTAR sequence with NC(12-55) containing NCp7 CCHC zinc fingers and flanking basic amino acid residues. From the chemical rates and the activation energy associated with the conformational fluctuations observed in the absence of NC, it is concluded that such fluctuations are associated with the opening and closing of the double-stranded terminal segments of cTAR. The destabilizing activity of NC(12-55) occurs mainly through a major increase of the opening rate constant of cTAR. Moreover, NC appears to augment the number of pathways between the open and closed states of cTAR, suggesting that it initiates melting of base-pairs at different locations within the terminal segments of cTAR. This activity of NC on the dynamics of cTAR secondary structure is thought to be critical for the formation of the cTAR-TAR complex, which is essential for the specificity and extent of proviral DNA synthesis by reverse transcriptase. PMID- 12581634 TI - Cis-acting regulatory elements in the potato virus X 3' non-translated region differentially affect minus-strand and plus-strand RNA accumulation. AB - The 72nt 3' non-translated region (NTR) of potato virus X (PVX) RNA is identical in all sequenced PVX strains and contains sequences that are conserved among all potexviruses. Computer folding of the 3' NTR sequence predicted three stem-loop structures (SL1, SL2, and SL3 in the 3' to 5' direction), which generally were supported by solution structure analyses. The importance of these sequence and/or structural elements to PVX RNA accumulation was further analyzed by inoculation of Nicotiana tabacum (NT-1) protoplasts with PVX transcripts containing mutations in the 3' NTR. Analyses of RNA accumulation by S(1) nuclease protection indicated that multiple sequence elements throughout the 3' NTR were important for minus strand RNA accumulation. Formation of SL3 was required for accumulation of minus strand RNA, whereas SL1 and SL2 formation were less important. However, sequences within all of these predicted structures were required for minus-strand RNA accumulation, including a conserved hexanucleotide sequence element in the loop of SL3, and the CU nucleotide in a U-rich sequence within SL2. In contrast, 13 nucleotides that were predicted to reside in SL1 could be deleted without any significant reduction in minus or plus-strand RNA levels. Potential polyadenylation signals (near upstream elements; NUEs) in the 3' NTR of PVX RNA were more important for plus-strand RNA accumulation than for minus-strand RNA accumulation. In addition, one of these NUEs overlapped with other sequence required for optimal minus-strand RNA levels. These data indicate that the PVX 3' NTR contains multiple, overlapping elements that influence accumulation of both minus and plus-strand RNA. PMID- 12581631 TI - Transcriptional profiling of Kruppel-like factor 4 reveals a function in cell cycle regulation and epithelial differentiation. AB - Kruppel-like factor 4 (KLF4) is an epithelially enriched, zinc finger-containing transcription factor, the expression of which is associated with growth arrest. Constitutive expression of KLF4 inhibits G1/S transition of the cell cycle but the manner by which it accomplishes this effect is unclear. To better understand the biochemical function of KLF4, we identified its target genes using cDNA microarray analysis in an established human cell line containing inducible KLF4. RNA extracted from induced and control cells were hybridized differentially to microarray chips containing 9600 human cDNAs. In all, 84 genes with significantly increased expression and 107 genes with significantly reduced expression due to KLF4 induction were identified. The affected genes are sorted to several clusters on the basis of functional relatedness. A major cluster belongs to genes involved in cell-cycle control. Within this cluster, many up-regulated genes are inhibitors of the cell cycle and down-regulated genes are promoters of the cell cycle. Another up-regulated gene cluster includes nine keratin genes, of which seven are located in a specific region on chromosome 12. The results indicate that KLF4 is involved in the control of cell proliferation and does so by eliciting changes in expression of numerous cell-cycle regulatory genes in a concerted manner. Furthermore, KLF4 regulates expression of a group of epithelial specific keratin genes in a manner consistent with a potential locus control region function. PMID- 12581635 TI - A Cdc25A antagonizing K vitamin inhibits hepatocyte DNA synthesis in vitro and in vivo. AB - Thioalkyl containing K vitamin analogs have been shown to be potent inhibitors of hepatoma cell growth and antagonizers of protein tyrosine phosphatase activity. We now show that they inhibit the activity of specific protein tyrosine phosphatases (PTP) in cell-free conditions in vitro, particularly the dual specificity phosphatase Cdc25A. Using primary cultures of adult rat hepatocytes that are in G0/G1 phase until stimulated into DNA synthesis by epidermal growth factor, we found that 2-(2-mercaptoethanol)-3-methyl-1,4-naphthoquinone or Compound 5 (Cpd 5) inhibited hepatocyte DNA synthesis and PTP activity in cell culture and in vivo after a two-thirds partial hepatectomy. We found a selective inhibition of Cdc25A activity in vitro, using both synthetic substrates and authentic cellular substrate, immunoprecipitated phospho-Cdk4. Intact Cpd 5 treated cells had decreased cellular Cdc25A activity and increased tyrosine phosphorylation of Cdk4, resulting in decreased phosphorylation of retinoblastoma (Rb). Loss of Cdk4 activity was confirmed using Cdk4 immunoprecipitates from either Cpd 5-treated or untreated cells and measuring its kinase activity using GST-Rb as target. We found a similar order of activity for inhibition of growth and Cdc25A activity using several thiol-containing analogs. Cdc25A inhibitors may thus be useful for defining biochemical pathways involving protein tyrosine phosphorylation that mediate cell growth inhibition. PMID- 12581636 TI - Topological measurement of an A-tract bend angle: comparison of the bent and straightened states. AB - It is well established that an A-tract imparts curvature to the DNA double helix. Constructs of such A-tracts have been used as bend standards in a large number of both structural and functional studies, and A-tracts can confer significant activation in transcription. An accurate value for the bend angle induced by an A tract is centrally important to all such studies, but the estimates reported for the bend angle of an A-tract differ by greater than threefold. To address this problem, we have used the rotational variant method to measure the angle of DNA curvature conferred by a tract of six adenine bases (A6 tract). The original version of the method measured a protein-induced bend angle independent of external standards. It compared the effect of bent and straight forms of the sequence on the topology of a DNA plasmid in which the sequence is cloned as a series of tandem repeats. To adapt the approach to the measurement of an intrinsic bend, high temperature was used to generate the straightened reference state, with the required topological relaxation being performed by a hyperthermophile topoisomerase. Appropriate plasmids containing tandem repeats of A-tracts were constructed and topologically analyzed in this manner. The bend value measured at 4 degrees C was 26(+/-2), and decreased linearly to 17(+/-2) at 37 degrees C. The relationship to other estimates and the application of these values are discussed. PMID- 12581637 TI - Topological measurement of an A-tract bend angle: variation of duplex winding. AB - The rotational variant method of Lutter et al. was developed to measure the bend angle induced when a protein binds to DNA. To measure the intrinsic bend conferred by a sequence of six adenine bases (an A6 tract), the method was modified by relaxing at high temperature to remove the bend. We describe here an alternative approach that involves unwinding the duplex DNA between adjacent bends in plasmids containing tandemly repeated blocks of A-tracts. This method measures the topological difference contributed by adjacent bends when they are in two different rotational settings, and therefore does not require reference to a straight state. The interbend DNA was unwound by use of the intercalator chloroquine, or, alternatively, by raising the temperature in the relaxation reaction. The effect of this unwinding is to change the pitch of the superhelix of the tandem repeats from which the bend angle is measured. The result is a bend angle value that is consistent with that measured using the bend-straightening version of the method. This version offers several advantages that complement the conventional bent versus straight approach. PMID- 12581638 TI - Crystal structure of activated ModE reveals conformational changes involving both oxyanion and DNA-binding domains. AB - ModE is a bacterial transcriptional regulator that orchestrates many aspects of molybdenum metabolism by binding to specific DNA sequences in a molybdate dependent fashion. We present the crystal structure of Escherichia coli ModE in complex with molybdate, which was determined at 2.75A from a merohedrally twinned crystal (twin fraction approximately 0.30) with space group P4(3). We now have structures of ModE in both its "switched on" (ligand-bound) and "switched off" (apo) states. Comparison with the apo structure shows that ligand binding leads to extensive conformational changes not only in the molybdate-binding domain, but also in the DNA-binding domain. The most obvious difference is the loss of the pronounced asymmetry between the two chains of the ModE dimer, which had been a characteristic property of the apo structure. Another major change concerns the relative orientation of the two DNA-interacting winged helix-turn-helix motifs. Manual docking of an idealized DNA structure suggests that this conformational change should improve DNA binding of the activated molybdate-bound ModE. PMID- 12581639 TI - Contractions and expansions of CAG/CTG trinucleotide repeats occur during ectopic gene conversion in yeast, by a MUS81-independent mechanism. AB - CAG/CTG trinucleotide repeat tracts expand and contract at a high rate during gene conversion in Saccharomyces cerevisiae. In order to characterize the mechanism responsible for such rearrangements, we built an experimental system based on the use of the rare cutter endonuclease I-SceI, to study the fate of trinucleotide repeat tracts during meiotic or mitotic (allelic or ectopic) gene conversion. After double-strand break (DSB) induced meiotic recombination, (CAG)(98) and (CAG)(255) are rearranged in 5% and 52% of the gene conversions, respectively, with similar proportions of contractions and expansions. No evidence of a meiotic hot spot activity associated with trinucleotide repeats could be found. When gene conversion is induced by a DSB during mitotic growth of the cells, no rearrangement of the repeat tracts is detected when the donor sequence is allelic to the recipient site of the DSB. However, when the donor sequence is at an ectopic location, frequent contractions and expansions of the repeat tract are found. No crossing-over associated with gene conversion could be detected. Mutants for the MUS81 gene, involved in the resolution of recombination intermediates, show a frequency of rearrangements identical with that of the wild type strain. We concluded that trinucleotide repeat rearrangements occur frequently during ectopic but not during allelic recombination, by a mechanism that does not require crossover formation. PMID- 12581640 TI - Enzyme catalysis via control of activation entropy: site-directed mutagenesis of 6,7-dimethyl-8-ribityllumazine synthase. AB - 6,7-Dimethyl-8-ribityllumazine synthase (lumazine synthase) catalyses the penultimate step in the biosynthesis of riboflavin. In Bacillus subtilis, 60 lumazine synthase subunits form an icosahedral capsid enclosing a homotrimeric riboflavin synthase unit. The ribH gene specifying the lumazine synthase subunit can be expressed in high yield. All amino acid residues exposed at the surface of the active site cavity were modified by PCR assisted mutagenesis. Polar amino acid residues in direct contact with the enzyme substrates, 5-amino-6 ribitylamino-2,4(1H,3H)-pyrimidinedione and 3,4-dihydroxy-2-butanone 4-phosphate, could be replaced with relative impunity with regard to the catalytic properties. Only the replacement of Arg127, which forms a salt bridge with the phosphate group of 3,4-dihydroxy-2-butanone 4-phosphate, reduced the catalytic rate by more than one order of magnitude. Replacement of His88, which is believed to assist in proton transfer reactions, reduced the catalytic activity by about one order of magnitude. Surprisingly, the activation enthalpy deltaH of the lumazine synthase reaction exceeds that of the uncatalysed reaction. On the other hand, the free energy of activation deltaG of the uncatalysed reaction is characterised by a large entropic term (TdeltaS) of -37.8 kJmol(-1), whereas the entropy of activation (TdeltaS) of the enzyme-catalysed reaction is -6.7 kJmol(-1). This suggests that the rate enhancement by the enzyme is predominantly achieved by establishing a favourable topological relation of the two substrates, whereas acid/base catalysis may play a secondary role. PMID- 12581642 TI - Control of directionality in L5 integrase-mediated site-specific recombination. AB - Mycobacteriophage L5 is a temperate phage that forms lysogens in Mycobacterium smegmatis. These lysogens carry an integrated L5 prophage inserted at a specific chromosomal location and undergo subsequent excision during induction of lytic growth. Both the integrative and excisive site-specific recombination events are catalyzed by the phage-encoded tyrosine integrase (Int-L5) and require the host encoded protein, mIHF. The directionality of these recombination events is determined by a second phage-encoded protein, Excise, the product of gene 36 (Xis L5); integration occurs efficiently in the absence of Xis-L5 while excision is dependent upon it. We show here that Xis-L5 binds to attR DNA, introduces a DNA bend, and facilitates the formation of an intasome-R complex. This complex, which requires mIHF, Xis-L5 and Int-L5, readily recombines with a second intasome formed by Int-L5, mIHF and attL DNA (intasome-L) to generate the attP and attB products of excision. Xis-L5 also strongly inhibits Int-L5-mediated integrative recombination but does not prevent either the protein-DNA interactions that form the attP intasome (intasome-P) or the capture of attB, but acts later in the reaction presumably by preventing the formation of a recombinagenic synaptic intermediate. The mechanism of action of Xis-L5 appears to be purely architectural, influencing the assembly of protein-DNA structures solely through its DNA-binding and DNA-bending properties. PMID- 12581641 TI - Involvement of retinoid X receptor alpha in coenzyme Q metabolism. AB - The nuclear retinoid X receptor alpha (RXRalpha) is the heterodimer partner in several nuclear receptors, some of them regulating lipid biosynthesis. Since coenzyme Q (CoQ) levels are greatly modified in aging and a number of diseases, we have investigated the involvement of RXRalpha in the biosynthetic regulation of this lipid by using a hepatocyte-specific RXRalpha-deficient mouse strain (RXRalpha-def). In the receptor-deficient liver, the amount of CoQ decreased to half of the control, and it was demonstrated that this decrease was caused by a significantly lowered rate of biosynthesis. On the other hand, induction of CoQ was extensive in both control and RXRalpha-def liver using the peroxisomal inducer di(2-ethylhexyl)phthalate (DEHP). Since the RXRalpha deficiency was specific to liver, no change in CoQ content or biosynthesis was observed in kidney. The other mevalonate pathway lipids, cholesterol and dolichol, were unchanged in the RXRalpha-def liver. Upon treatment with DEHP, cholesterol decreased in the control but remained unchanged in the receptor-deficient mice. In control mice, cold exposure elevated CoQ levels by 60%, but this induction did not occur in the liver of RXRalpha-def mice. In contrast, PPARalpha-null mice, which lack induction upon treatment with peroxisomal inducers, respond to cold exposure and CoQ content is increased. The amount of cholesterol decreased in both control and RXRalpha-def liver upon cold treatment. The results demonstrate that RXRalpha is required for CoQ biosynthesis and for its induction upon cold treatment, but does not appear to be involved in the basic synthesis of cholesterol and dolichol. The receptor is not involved in the elevated CoQ biosynthesis during peroxisomal induction. PMID- 12581643 TI - Structure of beta-antithrombin and the effect of glycosylation on antithrombin's heparin affinity and activity. AB - Antithrombin is a member of the serpin family of protease inhibitors and the major inhibitor of the blood coagulation cascade. It is unique amongst the serpins in that it circulates in a conformation that is inactive against its target proteases. Activation of antithrombin is brought about by a conformational change initiated upon binding heparin or heparan sulphate. Two isoforms exist in the circulation, alpha-antithrombin and beta-antithrombin, which differ in the amount of glycosylation present on the polypeptide chain; beta-antithrombin lacks the carbohydrate present at Asn135 in alpha-antithrombin. Of the two forms, beta antithrombin has the higher affinity for heparin and thus functions as the major inhibitor in vivo even though it is the less abundant form. The reason for the differences in heparin affinity between the alpha and beta-forms have been shown to be due to the additional carbohydrate changing the rate of the conformational change. Here, we describe the most accurate structures of alpha-antithrombin and alpha-antithrombin+heparin pentasaccharide reported to date (2.6A and 2.9A resolution, respectively, both re-refinements using old data), and the structure of beta-antithrombin (2.6A resolution). The new structures have a remarkable degree of ordered carbohydrate and include parts of the antithrombin chain not modeled before. The structures have allowed a detailed comparison of the conformational differences between the three. They show that the structural basis of the lower affinity for heparin of alpha-antithrombin over beta-antithrombin is due to the conformational change that occurs upon heparin binding being sterically hindered by the presence of the additional bulky carbohydrate at Asn135. PMID- 12581644 TI - Two carbohydrate binding sites in the H(CC)-domain of tetanus neurotoxin are required for toxicity. AB - Tetanus neurotoxin binds via its carboxyl-terminal H(C)-fragment selectively to neurons mediated by complex gangliosides. We investigated the lactose and sialic acid binding pockets of four recently discovered potential binding sites employing site-directed mutagenesis. Substitution of residues in the lactose binding pocket drastically decreased the binding of the H(C)-fragment to immobilized gangliosides and to rat brain synaptosomes as well as the inhibitory action of recombinant full length tetanus neurotoxin on exocytosis at peripheral nerves. The conserved motif of S(1287)XWY(1290) em leader G(1300) assisted by N1219, D1222, and H1271 within the lactose binding site comprises a typical sugar binding pocket, as also present, for example, in cholera toxin. Replacement of the main residue of the sialic acid binding site, R1226, again caused a dramatic decline in binding affinity and neurotoxicity. Since the structural integrity of the H(C)-fragment mutants was verified by circular dichroism and fluorescence spectroscopy, these data provide the first biochemical evidence that two carbohydrate interaction sites participate in the binding and uptake process of tetanus neurotoxin. The simultaneous binding of one ganglioside molecule to each of the two binding sites was demonstrated by mass spectroscopy studies, whereas ganglioside-mediated linkage of native tetanus neurotoxin molecules was ruled out by size exclusion chromatography. Hence, a subsequent displacement of one ganglioside by a glycoprotein receptor is discussed. PMID- 12581645 TI - Structural basis for the interaction between the Tap/NXF1 UBA domain and FG nucleoporins at 1A resolution. AB - The mRNA nuclear export function of Tap/NXF1 requires interactions with nuclear pore proteins (nucleoporins) that contain characteristic Phe-Gly repeats based on FG, GLFG or FxFG cores separated by hydrophilic linkers. FG-nucleoporins bind the two most C-terminal domains of Tap, which have NTF2 and UBA folds, respectively. We used a combination of NMR and X-ray crystallography to define the interaction interface between Tap UBA and FxFG nucleoporins and show that it involves primarily the two aromatic rings of the FxFG core that bind in a hydrophobic surface depression centred on Tap Cys588. NMR evidence indicates that the same depression mediates the binding of GLFG nucleoporins, which we confirmed by demonstrating competition between the two classes of repeat for binding to Tap UBA. Moreover, modification of Cys588 reduced the binding of Tap UBA to both GLFG and FxFG nucleoporins as well as to nuclear envelopes. These data underscore the central role of the conserved FG-nucleoporin repeat cores in binding to Tap UBA and indicate that functional differences between different classes of nucleoporins depend more on their spatial distribution in nuclear pores than on their binding to different sites on Tap UBA. PMID- 12581646 TI - The Oligomeric states of Haloarcula marismortui malate dehydrogenase are modulated by solvent components as shown by crystallographic and biochemical studies. AB - The three-dimensional crystal structure of the (R207S, R292S) mutant of malate dehydrogenase from Haloarcula marismortui was solved at 1.95A resolution in order to determine the role of salt bridges and solvent ions in halophilic adaptation and quaternary structure stability. The mutations, located at the dimer-dimer interface, disrupt two inter-dimeric salt bridge clusters that are essential for wild-type tetramer stabilisation. Previous experiments in solution, performed on the double mutant, had shown a tetrameric structure in 4M NaCl, which dissociated into active dimers in 2M NaCl. In order to establish if the active dimeric form is a product of the mutation, or if it also exists in the wild-type protein, complementary studies were performed on the wild-type enzyme by analytical centrifugation and small angle neutron scattering experiments. They showed the existence of active dimers in NaF, KF, Na(2)SO(4), even in the absence of NADH, and in the presence of NADH at concentrations of NaCl below 0.3M. The crystal structure shows a tetramer that, in the absence of the salt bridge clusters, appears to be stabilized by a network of ordered water molecules and by Cl(-) binding at the dimer-dimer interface. The double mutant and wild-type dimer folds are essentially identical (the r.m.s. deviation between equivalent C(alpha) positions is 0.39A). Chloride ions are also observed at the monomer-monomer interfaces of the mutant, contributing to the stability of each dimer against low salt dissociation. Our results support the hypothesis that extensive binding of water and salt is an important feature of adaptation to a halophilic environment. PMID- 12581647 TI - Crystal structure of Stefin A in complex with cathepsin H: N-terminal residues of inhibitors can adapt to the active sites of endo- and exopeptidases. AB - Binding of cystatin-type inhibitors to papain-like exopeptidases cannot be explained by the stefin B-papain complex. The crystal structure of human stefin A bound to an aminopeptidase, porcine cathepsin H, has been determined in monoclinic and orthorhombic crystal forms at 2.8A and 2.4A resolutions, respectively. The asymmetric unit of each form contains four complexes. The structures are similar to the stefin B-papain complex, but with a few distinct differences. On binding, the N-terminal residues of stefin A adopt the form of a hook, which pushes away cathepsin H mini-chain residues and distorts the structure of the short four residue insertion (Lys155A-Asp155D) unique to cathepsin H. Comparison with the structure of isolated cathepsin H shows that the rims of the cathepsin H structure are slightly displaced (up to 1A) from their position in the free enzyme. Furthermore, comparison with the stefin B-papain complex showed that molecules of stefin A bind about 0.8A deeper into the active site cleft of cathepsin H than stefin B into papain. The approach of stefin A to cathepsin H induces structural changes along the interaction surface of both molecules, whereas no such changes were observed in the stefin B-papain complex. Carboxymethylation of papain seems to have prevented the formation of the genuine binding geometry between a papain-like enzyme and a cystatin-type inhibitor as we observe it in the structure presented here. PMID- 12581648 TI - Localization of the trigger factor binding site on the ribosomal 50S subunit. AB - In Escherichia coli, protein folding is undertaken by three distinct sets of chaperones, the DnaK-DnaJ and GroEL-GroES systems and the trigger factor (TF). TF has been proposed to be the first chaperone to interact with the nascent polypeptide chain as it emerges from the tunnel of the 70S ribosome and thus probably plays an important role in co-translational protein folding. We have made complexes with deuterated ribosomes (50S subunits and 70S ribosomes) and protated TF and determined the TF binding site on the respective complexes using the neutron scattering technique of spin-contrast variation. Our data suggest that the TF binds in the form of a homodimer. On both the 50S subunit and the 70S ribosome, the TF position is in proximity to the tunnel exit site, near ribosomal proteins L23 and L29, located on the back of the 50S subunit. The positions deviate from one another, such that the position on the 70S ribosome is located slightly further from the tunnel than that determined for the 50S subunit alone. Nevertheless, from both determined positions interaction between TF and a short nascent chain of 57 amino acid residues would be plausible, compatible with a role for TF participation in co-translational protein folding. PMID- 12581649 TI - Solution structure of switch Arc, a mutant with 3(10) helices replacing a wild type beta-ribbon. AB - Adjacent N11L and L12N mutations in the antiparallel beta-ribbon of Arc repressor result in dramatic changes in local structure in which each beta-strand is replaced by a right-handed helix. The full solution structure of this "switch" Arc mutant shows that irregular 3(10) helices compose the new secondary structure. This structural metamorphosis conserves the number of main-chain and side-chain to main-chain hydrogen bonds and the number of fully buried core residues. Apart from a slight widening of the interhelical angle between alpha helices A and B and changes in side-chain conformation of a few core residues in Arc, no large-scale structural adjustments in the remainder of the protein are necessary to accommodate the ribbon-to-helix change. Nevertheless, some changes in hydrogen-exchange rates are observed, even in regions that have very similar structures in the two proteins. The surface of switch Arc is packed poorly compared to wild-type, leading to approximately 1000A(2) of additional solvent accessible surface area, and the N termini of the 3(10) helices make unfavorable head-to-head electrostatic interactions. These structural features account for the positive m value and salt dependence of the ribbon-to-helix transition in Arc N11L, a variant that can adopt either the mutant or wild-type structures. The tertiary fold is capped in different ways in switch and wild-type Arc, showing how stepwise evolutionary transformations can arise through small changes in amino acid sequence. PMID- 12581650 TI - Solvation effects and driving forces for protein thermodynamic and kinetic cooperativity: how adequate is native-centric topological modeling? AB - What energetic and solvation effects underlie the remarkable two-state thermodynamics and folding/unfolding kinetics of small single-domain proteins? To address this question, we investigate the folding and unfolding of a hierarchy of continuum Langevin dynamics models of chymotrypsin inhibitor 2. We find that residue-based additive Go-like contact energies, although native-centric, are by themselves insufficient for protein-like calorimetric two-state cooperativity. Further native biases by local conformational preferences are necessary for protein-like thermodynamics. Kinetically, however, even models with both contact and local native-centric energies do not produce simple two-state chevron plots. Thus a model protein's thermodynamic cooperativity is not sufficient for simple two-state kinetics. The models tested appear to have increasing internal friction with increasing native stability, leading to chevron rollovers that typify kinetics that are commonly referred to as non-two-state. The free energy profiles of these models are found to be sensitive to the choice of native contacts and the presumed spatial ranges of the contact interactions. Motivated by explicit water considerations, we explore recent treatments of solvent granularity that incorporate desolvation free energy barriers into effective implicit-solvent intraprotein interactions. This additional feature reduces both folding and unfolding rates vis-a-vis that of the corresponding models without desolvation barriers, but the kinetics remain non-two-state. Taken together, our observations suggest that interaction mechanisms more intricate than simple Go-like constructs and pairwise additive solvation-like contributions are needed to rationalize some of the most basic generic protein properties. Therefore, as experimental constraints on protein chain models, requiring a consistent account of protein like thermodynamic and kinetic cooperativity can be more stringent and productive for some applications than simply requiring a model heteropolymer to fold to a target structure. PMID- 12581651 TI - Interplay among tertiary contacts, secondary structure formation and side-chain packing in the protein folding mechanism: all-atom representation study of protein L. AB - Experimental and theoretical results suggest that, since proteins are energetically minimally frustrated, the native fold, or topology, plays a primary role in determining the structure of the transition state ensemble and on-pathway intermediate states in protein folding. Although the central role of native state topology in determining the folding mechanism is thought to be a quite general result-at least for small two-state folding proteins-there are remarkable exceptions. Recent experimental findings have shown that topology alone cannot always determine the folding mechanism, and demonstrated that the balance between topology and energetics is very delicate. This balance seems to be particularly critical in proteins with a highly symmetrical native structure, such as proteins L and G, which have similar native structure topology but fold by different mechanisms. Simplified, C(alpha)-atom only protein models have shown not be sufficient to differentiate these mechanisms. An all-atom Go model provides a valuable intermediate model between structurally simplified protein representations and all-atom protein simulations with explicit/implicit solvent descriptions. We present here a detailed study of an all-atom Go-like representation of protein L, in close comparison with the experimental results and with the results obtained from a simple C(alpha)-atom representation of the same protein. We also perform simulations for protein G, where we obtain a folding mechanism in which the protein symmetry is broken exactly in the opposite way to protein L as has been observed experimentally. A detailed analysis for protein L also shows that the role of specific residues is correctly and quantitatively reproduced by the all-atom Go model over almost the entire protein. PMID- 12581652 TI - Functional sites in protein families uncovered via an objective and automated graph theoretic approach. AB - We report a method for detection of recurring side-chain patterns (DRESPAT) using an unbiased and automated graph theoretic approach. We first list all structural patterns as sub-graphs where the protein is represented as a graph. The patterns from proteins are compared pair-wise to detect patterns common to a protein pair based on content and geometry criteria. The recurring pattern is then detected using an automated search algorithm from the all-against-all pair-wise comparison data of proteins. Intra-protein pattern comparison data are used to enable detection of patterns recurring within a protein. A method has been proposed for empirical calculation of statistical significance of recurring pattern. The method was tested on 17 protein sets of varying size, composed of non-redundant representatives from SCOP superfamilies. Recurring patterns in serine proteases, cysteine proteases, lipases, cupredoxin, ferredoxin, ferritin, cytochrome c, aspartoyl proteases, peroxidases, phospholipase A2, endonuclease, SH3 domain, EF hand and lectins show additional residues conserved in the vicinity of the known functional sites. On the basis of the recurring patterns in ferritin, EF-hand and lectins, we could separate proteins or domains that are structurally similar yet different in metal ion-binding characteristics. In addition, novel recurring patterns were observed in glutathione-S-transferase, phospholipase A2 and ferredoxin with potential structural/functional roles. The results are discussed in relation to the known functional sites in each family. Between 2000 and 50,000 patterns were enumerated from each protein with between ten and 500 patterns detected as common to an evolutionarily related protein pair. Our results show that unbiased extraction of functional site pattern is not feasible from an evolutionarily related protein pair but is feasible from protein sets comprising five or more proteins. The DRESPAT method does not require a user-defined pattern, size or location of the pattern and therefore, has the potential to uncover new functional sites in protein families. PMID- 12581654 TI - Histone chaperones and nucleosome assembly. AB - Recent structures of the nucleosome core particle reveal details of histone histone and histone-DNA interactions. These structures have now set the stage for understanding chromatin assembly and dynamics during replication and transcription. Histone chaperones and chromatin remodeling complexes are important in both of these processes. The nucleosome and its protein core, the histone octamer, have twofold symmetry, which histone chaperones may use to bind core histones. Recent studies suggest that the nucleoplasmin pentamer may mediate histone storage, sperm chromatin decondensation and nucleosome assembly, by dimerizing to form a decamer. In this model, histone binding on the lateral surface of the chaperone involves stereospecific interactions and a shared twofold axis. PMID- 12581655 TI - DNA manipulators: caught in the act. AB - DNA is a dynamic molecule that undergoes constant changes in the cell through interactions with numerous proteins. Several classes of enzyme are specialized in promoting DNA rearrangements, including site-specific recombinases, DNA helicases, transposases and DNA topoisomerases. Recent structures of protein-DNA reaction intermediates trapped in various states of DNA remodeling, complemented by biochemical and biophysical functional studies, have enhanced our understanding of their respective mechanistic pathways. PMID- 12581656 TI - Damage repair DNA polymerases Y. AB - The newly found Y-family DNA polymerases are characterized by low fidelity replication using an undamaged template and the ability to carry out translesion DNA synthesis. The crystal structures of three Y-family polymerases, alone or complexed with DNA and nucleotide substrate, reveal a conventional right-hand like catalytic core consisting of finger, thumb and palm domains. The finger and thumb domains are unusually small resulting in an open and spacious active site, which can accommodate mismatched base pairs as well as various DNA lesions. Although devoid of a 3'-->5' exonuclease activity, the Y-family polymerases possess a unique "little finger" domain that facilitates DNA association, catalytic efficiency and interactions with auxiliary factors. Expression of Y family polymerases is often induced by DNA damage, and their recruitment to the replication fork is mediated by beta-clamp, clamp loader, single-strand-DNA binding protein and RecA in Escherichia coli, and by ubiquitin-modified proliferating cell nuclear antigen in yeast. PMID- 12581657 TI - Bacterial RNA polymerases: the wholo story. AB - Recent structural and biophysical results have provided unprecedented insights into the structure and function of the bacterial RNA polymerase holoenzyme as it goes through the steps of transcription initiation. Comparisons with structural analyses of evolutionarily unrelated RNA polymerases reveal unexpected general features of the initiation process. PMID- 12581658 TI - Eukaryotic transcriptional regulatory complexes: cooperativity from near and afar. AB - It is characteristic of eukaryotic transcription that a unique combination of multiple transcriptional regulatory proteins bound to promoter DNA specifically activate or repress downstream target genes; this is referred to as combinatorial gene regulation. Recently determined structures have revealed different modes of protein-protein interaction on the promoter DNA from near (e.g. the Runx1-CBFbeta DNA, NFAT-Fos-Jun-DNA, GABPalpha-GABPbeta-DNA, Ets-1-Pax-5-DNA and PU.1-IRF-4-DNA complexes) and afar with DNA looping (e.g. the c-Myb-C/EBPbeta-DNA complex), and their regulatory mechanisms. PMID- 12581659 TI - RNA-modifying enzymes. AB - A bewildering number of post-transcriptional modifications are introduced into cellular RNAs by enzymes that are often conserved among archaea, bacteria and eukaryotes. The modifications range from those with well-understood functions, such as tRNA aminoacylation, to widespread but more mysterious ones, such as pseudouridylation. Recent structure determinations have included two types of RNA nucleobase modifying enzyme: pseudouridine synthases and tRNA guanine transglycosylases. PMID- 12581660 TI - Eukaryotic translation initiation factors and regulators. AB - Significant progress has been made over the past several years on structural studies of the eukaryotic translation initiation factors that facilitate the assembly of a translation-competent ribosome at the initiation codon of an mRNA. These structural studies have revealed the repeated use of a set of common structural folds, highlighted the evolutionary conservation of the translation apparatus, and provided insight into the mechanism and regulation of cellular and viral protein synthesis. PMID- 12581661 TI - S-domain assembly of the signal recognition particle. AB - The signal recognition particle (SRP) is a phylogenetically conserved ribonucleoprotein that associates with ribosomes to mediate the targeting of membrane and secretory proteins to biological membranes. In higher eukaryotes, SRP biogenesis involves the sequential binding of SRP19 and SRP54 proteins to the S domain of 7S RNA. The recently determined crystal structures of SRP19 in complex with the S domain, and that of the ternary complex of SRP19, the S domain and the M domain of SRP54, provide insight into the molecular basis of S-domain assembly and SRP function. PMID- 12581663 TI - Early events in protein folding. AB - Recent advances have significantly increased the time and spectroscopic resolution of protein folding experiments. We can now study the timescale and nature of polypeptide collapse, and how this correlates with secondary and tertiary structure formation. Studies on ultrafast folding proteins and peptides provide experimental benchmarks on a timescale that overlaps directly with that of molecular dynamics simulations. This makes possible direct tests of both simulations and current models of protein folding. PMID- 12581664 TI - Protein folding: bringing theory and experiment closer together. AB - The ability to perform enzymatic function by harnessing random molecular motion into self-organized protein structures is one of the most fascinating results of evolution. A close interplay between theory and experiment is driving the progress in understanding the principles that determine the behaviour of proteins. New techniques that significantly increase the amount of information obtainable from experimental data have been recently proposed; it is now becoming possible to describe at atomic resolution the events that take place during the folding process. Successful predictions of these events are being reported at an increasing rate and general principles are being outlined. PMID- 12581665 TI - Single-molecule folding. AB - Recent developments in fluorescence and force spectroscopy enable us to go beyond the ensemble average and measure the behavior of individual biomacromolecules. These single-molecule approaches can directly resolve transient intermediate states and multiple reaction pathways, and thus are uniquely powerful in characterizing the complex dynamics of biological processes. Recent applications of these two techniques to the protein and RNA folding problems have led to exciting new results. PMID- 12581666 TI - Protein unfolding--an important process in vivo? AB - Protein unfolding is an important step in several cellular processes, most interestingly protein degradation by ATP-dependent proteases and protein translocation across some membranes. Unfolding can be catalyzed when the unfoldases change the unfolding pathway of substrate proteins by pulling at their polypeptide chains. The resistance of a protein to unraveling during these processes is not determined by the protein's stability against global unfolding, as measured by temperature or solvent denaturation in vitro. Instead, resistance to unfolding is determined by the local structure that the unfoldase encounters first as it follows the substrate's polypeptide chain from the targeting signal. As unfolding is a necessary step in protein degradation and translocation, the susceptibility to unfolding of substrate proteins contributes to the specificity of these important cellular processes. PMID- 12581667 TI - Evolutionary aspects of protein structure and folding. AB - The traditional reconstruction of molecular events of the past based on sequence conservation becomes very vague beyond one to two billion years ago. There are certain molecular features, however, such as polymer flexibility and loop closure, that are conserved merely because of their physical nature. This allows one to penetrate the earliest stages of protein evolution. PMID- 12581668 TI - Applications of calorimetric methods to drug discovery and the study of protein interactions. AB - Recent studies report the application of isothermal titration calorimetry and differential scanning calorimetry to the study of protein-ligand interactions, allosteric cooperativity and aspects of protein folding. New methods of data analysis compare alternative methods for determining ligand binding enthalpy and analyze potential sources of error in the experimental measurement of other thermodynamic parameters. Several reports examine issues concerning drug design and the correlation of thermodynamic and X-ray structural data. New instruments allow volumetric effects in biochemical systems to be evaluated calorimetrically and to substantially expand the throughput of differential scanning calorimetry measurements in drug discovery and other high-throughput applications. PMID- 12581669 TI - Ras-effector interactions: after one decade. AB - Ras effectors have convergently developed a common subdomain in their otherwise unrelated protein body for their interaction with Ras. Structural analysis revealed that the mode of interaction is highly similar for all Ras effectors, but is completely different from that of effectors of other subfamilies of small GTPases. Whereas the molecular mechanism of effector activation is still elusive, detailed knowledge about the thermodynamics and dynamics of the interaction with Ras has accumulated. PMID- 12581670 TI - Additivity-based prediction of equilibrium constants for some protein-protein associations. AB - For many protein families, such as serine proteinases or serine proteinase inhibitors, the family assignment predicts reactivity only in general terms. Both detailed specificity and quantitative reactivity are lacking. We believe that, for many such protein families, algorithms can be devised by defining the subset of n functionally important sequence positions, making the 19n possible single mutants and measuring their reactivity. Given the assumption that the contributions of the n positions are additive, the reactivities of the 20(n) variants can be predicted. This is illustrated by an almost complete algorithm for the Kazal family of protein inhibitors of serine proteinases. PMID- 12581674 TI - Atheromatous plaque location and arterial remodelling. PMID- 12581675 TI - The role of the endothelin system in myocardial infarction--new therapeutic targets? PMID- 12581676 TI - Pulmonary arterial hypertension and the vasoconstrictive factor: is there still a role for vasodilator testing? PMID- 12581677 TI - Manpower in cardiology II in western and central Europe (1999-2000). PMID- 12581678 TI - Trends in coronary events in Finland during 1983-1997. The FINAMI study. AB - AIMS: To analyse the trends in incidence, recurrence, case fatality, and treatments of acute coronary events in Finland during the 15-year period 1983-97. METHODS AND RESULTS: Population-based MI registration has been carried out in defined geographical areas, first as a part of the FINMONICA Project and then continued as the FINAMI register. During the study period, 6501 coronary heart disease (CHD) events were recorded among men and 1778 among women aged 35-64 years. The CHD mortality declined on average 6.4%/year (95% confidence interval 5.4, -7.4%) among men and 7.0%/year (-4.7, -9.3%) among women. The mortality from recurrent events declined even more steeply, 9.9%/year (-8.3, -11.4%) among men and 9.3%/year (-5.1, -13.4%) among women. The proportion of recurrent events of all CHD events also declined significantly in both sexes. Of all coronary deaths, 74% among men and 61% among women took place out-of-hospital. The decline in 28 day case fatality was 1.3%/year (-0.3, -2.3%) among men and 3.1%/year (-0.7, 5.5%) among women. CONCLUSIONS: The study period was characterized by a marked reduction in the occurrence of recurrent CHD events and a relatively modest reduction in the 28-day case fatality. The findings suggest that primary and secondary prevention have played the main roles in the decline in CHD mortality in Finland. PMID- 12581680 TI - Eccentric atherosclerotic plaques with positive remodelling have a pericardial distribution: a permissive role of epicardial fat? A three-dimensional intravascular ultrasound study of left anterior descending artery lesions. AB - AIMS: The transversal distribution of coronary atherosclerotic plaques (AP) (myocardial vs pericardial) affects vessel remodelling. The aim of this study was to define the impact of transversal lesion distribution on vessel remodelling in proximal and distal coronary segments using a 3D intravascular ultrasound (IVUS) reconstruction. METHODS: The study group included 70 lesions located in the left anterior descending artery within 5mm of the septal take-off, and imaged using 3D IVUS. The take-off of the septal branch was used to divide the plaque into a myocardial and pericardial surface. The IVUS index of vessel remodelling was calculated as: [narrowest external elastic membrane (EEM) site cross-sectional area (CSA)-reference EEM CSA)/reference EEM CSAx100]. The lesions with an intermediate vessel remodelling index (between -25% and +15%) were excluded from analysis. RESULTS: Of the 38 APs with a pericardial distribution, 34 (89%) showed positive remodelling (P<0.001). The distal lesions had a positive vessel remodelling index regardless of transversal plaque distribution. At multivariate analysis, pericardial distribution and the distal location of AP were the only independent variables predictive of positive remodelling. CONCLUSIONS: The transversal distribution of atherosclerotic plaque affects vessel remodelling in left anterior descending coronary lesions, probably because of an extravascular splinting effect. Distal lesions usually show positive remodelling regardless of transversal plaque distribution. PMID- 12581679 TI - Prevalence of left ventricular diastolic dysfunction in the community. Results from a Doppler echocardiographic-based survey of a population sample. AB - AIMS: The prevalence of left ventricular diastolic abnormalities in the general population is largely unclear. Thus, the aim of this study was, firstly, to identify abnormal diastolic function by echocardiography in an age-stratified population-based European sample (MONICA Augsburg, n=1274, 25 to 75 years, mean 51+/-14) and, secondly, to analyse clinical and anthropometric parameters associated with diastolic abnormalities. METHODS AND RESULTS: The overall prevalence of diastolic abnormalities, as defined by the European Study Group on Diastolic Heart Failure (i.e. age dependent isovolumic relaxation time (92-105 ms) and early (E-wave) and late (A-wave) left ventricular filling (E/A-ratio, 1 0.5)) was 11.1%. When only subjects treated with diuretics or with left atrial enlargement were considered (suggesting diastolic dysfunction) the prevalence was 3.1%. The prevalence of diastolic abnormalities varied according to age: from 2.8% in individuals aged 25-35 years to 15.8% among those older than 65 years (P<0.01). Significantly higher rates of diastolic abnormalities were observed in men as compared to women (13.8% vs 8.6%, P<0.01). Independent predictors of diastolic abnormalities were arterial hypertension, evidence of left ventricular (LV) hypertrophy, and coronary artery disease. Interestingly, in the absence of these predisposing conditions, diastolic abnormalities (4.3%) or diastolic dysfunction (1.1%) were rare, even in subjects older than 50 years of age (4.6%) and (1.2%), respectively. In addition to these factors, diastolic dysfunction was related to high body mass index, high body fat mass, and diabetes mellitus. CONCLUSION: The prevalences of diastolic abnormalities and diastolic dysfunction are higher than that of systolic dysfunction and are increased (despite age dependent diagnostic criteria) in the elderly. However, in the absence of risk factors for diastolic abnormalities or diastolic dysfunction, namely LV hypertrophy, arterial hypertension, coronary artery disease, obesity and diabetes the condition is rare even in elderly subjects. These data allow speculation on whether diastolic heart failure may be prevented by improved implementation of measures directed against predisposing conditions. PMID- 12581681 TI - Prognostic value of ST-segment resolution-when and what to measure. AB - AIMS: Analyses of ST-segment resolution during acute myocardial infarction has, during recent years, challenged coronary angiography as gold-standard for predicting myocardial reflow and future risk. We have previously reported that continuous ST-monitoring can be done accurately in the clinical setting. We now set out to compare the prognostic value of previously suggested cut-offs for ST segment resolution, and determine the times to measure these. METHODS AND RESULTS: We analysed 752 patients with ST-elevation infarction, from the second Assessment of Safety and Efficacy of a New Thrombolytic (ASSENT 2) and ASSENT PLUS studies, either with vectorcardiography or continuous 12-lead ST-monitoring. All analyses were made blindly by two independent observers. Times to 20, 30, 50 and 70% ST-segment resolution were examined in relation to 30-day mortality.The optimal cut-off for ST-segment resolution analyses was found to be 50%, measured at 60 min. We could hereby identify a large low-risk group, 40% of the population, with only 1.4% 30-day mortality. Furthermore, 88% of deaths were correctly predicted within 1h of observation and treatment. CONCLUSION: Continuous ST-monitoring of patients with acute myocardial infarction yields important prognostic information after 60 min of observation and should be used for very early-risk stratification in these patients. PMID- 12581682 TI - Relationship between transcardiac gradient of endothelin-1 and left ventricular remodelling in patients with first anterior myocardial infarction. AB - Aims To evaluate whether plasma endothelin-1 (ET-1) is extracted or produced through the heart in patients with acute myocardial infarction (AMI), and the relationship between transcardiac extraction of plasma ET-1 and left ventricular (LV) remodelling. Methods and results We measured the plasma level of ET-1 in the aortic root (Ao) and coronary sinus (CS) in 48 consecutive patients, who received successful revascularization and enalapril, for a first anterior AMI. In the acute phase the plasma ET-1 level was significantly higher both in the Ao and the CS compared to the control subjects. However, the plasma ET-1 level was significantly lower in the CS than in the Ao in the acute phase and after 1 month. There were significant correlations between transcardiac extraction of ET 1 in the acute phase and LV ejection fraction and LV end-diastolic volume index (LVEDVI) after 1 month. Stepwise multivariate analysis showed that maximal creatine phosphokinase and transcardiac extraction of plasma ET-1 during the acute phase were independently and positively correlated with the absolute change in LVEDVI after 1 month. Conclusions These results indicate that elevated circulating ET-1 is extracted through the heart in patients with a first anterior AMI and that the extracted ET-1 plays a significant role in modulating post infarct LV remodelling. PMID- 12581683 TI - Assessment of the vasodilator response in primary pulmonary hypertension. Comparing prostacyclin and iloprost administered by either infusion or inhalation. AB - AIMS: To directly compare the differential effects of oxygen, prostacyclin and iloprost (aerosolized and intravenous) in primary pulmonary hypertension. METHODS AND RESULTS: Twenty-one patients with severe primary pulmonary hypertension underwent right heart catheterization following oxygen inhalation, inhalation of aerosolized iloprost, intravenous prostacyclin or intravenous iloprost. The stability of the iloprost solution was tested for up to 4 weeks. Oxygen slightly decreased pulmonary vascular resistance. Intravenous prostacyclin (7.2+/-3.4 ng kg(-1) min(-1)) reduced pulmonary (1772+/-844 vs 1325+/-615 dyn s cm(-5), P<0.001) and systemic vascular resistance, and arterial and right atrial pressure, while cardiac output increased. Iloprost inhalation diminished pulmonary (1813+/-827 vs 1323+/-614 dyn s cm(-5), P<0.001) and systemic vascular resistance, and pulmonary artery (58+/-12 vs 50+/-12 mmHg,P<0.001) and right atrial pressure, while cardiac output increased. With intravenous iloprost (1.2+/ 0.5 ng kg(-1) min(-1), n=8) a decrease in pulmonary (2202+/-529 vs 1515+/-356 dyn s cm(-5), P<0.05) and systemic vascular resistance and right a trial pressure occurred while cardiac output increased. Iloprost solution remained stable for 33 days while losing <10% (4 degrees C) of its active drug concentration.Conclusions Intravenous iloprost and prostacyclin have very similar haemodynamic profiles. In contrast, only inhaled iloprost exerted selective pulmonary vasodilation, reducing pulmonary vascular resistance and pulmonary artery pressure without systemic vasodilation. The longer half-life and extended stability despite lower costs render iloprost an attractive alternative to chronic prostacyclin treatment in primary pulmonary hypertension. PMID- 12581684 TI - Novel management strategy for patients with suspected pulmonary embolism. AB - AIMS: A simple management strategy is required for patients with acute pulmonary embolism which allows a rapid and reliable diagnosis in order to start timely and appropriate treatment. METHODS AND RESULTS: Two hundred and four consecutive patients with suspected pulmonary embolism were managed according to a standardized protocol based on the clinical pretest probability and the initial haemodynamic presentation (shock index=heart rate divided by systolic blood pressure). Patients with a high pretest probability and a positive shock index (> or =1) (n=21) underwent urgent transthoracic echocardiography. Based on the presence or absence of right ventricular dysfunction, reperfusion treatment was initiated immediately. Patients with a negative shock index (<1) (n=183) underwent diagnostic evaluation including pretest probability, D-dimer, and spiral computed tomography (CT) as first-line tests. Echocardiography was performed only when a central pulmonary embolism was found in the spiral CT(n=33). According to our strategy, 98 patients met the diagnostic criteria of pulmonary embolism: 75 patients (all shock index <1) were treated with heparin alone, 16 (seven had a shock index > or =1) with thrombolysis, four (all shock index > or =1) with catheter fragmentation, and three (all shock index > or =1) with surgical embolectomy. The all-cause mortality rate at 30 days was 5%, and at 6 months 11%. Right ventricular dysfunction on baseline echocardiography was not associated with a higher mortality rate at 6 months (logrank 2.4, P=0.12). CONCLUSIONS: The novel management strategy for patients with suspected pulmonary embolism resulted in a rapid diagnosis and treatment with a low 30-day mortality. In patients with pulmonary embolism and a positive shock index, time-consuming imaging tests can be avoided to reduce the risk of sudden death and not to delay reperfusion therapy. PMID- 12581685 TI - An investigation into the relationship between salivary cortisol, stress, anxiety and depression. AB - This study examined the relationship between indices of self-reported emotional distress and absolute versus change in cortisol levels. Fifty-four women attending a diagnostic breast clinic completed scales measuring stress, anxiety and depression and provided five saliva samples over the course of a single day for the measurement of cortisol. No significant relationships were evident between absolute cortisol levels and the distress measures. Analysis of the change in cortisol levels revealed a non-linear interaction effect between stress and anxiety and time of day. There was a non-linear relation between time of day and cortisol levels, but the extent of the non-linearity was dependent upon levels of stress and anxiety, not depression. A relationship was apparent between indices of distress and change in cortisol levels, but not absolute levels of the hormone. PMID- 12581686 TI - Implicit memory bias and trait anxiety: a psychophysiological analysis. AB - The effect of threatening words and anxiety upon implicit memory performance was investigated. It was predicted that anxious individuals would show a bias to threat-related material. In addition, psychophysiological measures were obtained to assess the attentional and encoding processes that might underlie this cognitive bias. Forty participants were equally allocated to high and low trait anxious groups, according to pre-determined cut-offs. All participants were exposed to threat and non-threat words and following a filler task, were asked to complete primed and unprimed wordstems. Implicit memory performance was assessed in terms of accuracy and reaction time for completion. Heart rate and electrodermal responses were measured. Results demonstrated initial increased cardiac deceleration to threat stimuli, subsequent cardiac acceleration to non threat stimuli, and an implicit memory bias to non-threat material by all participants. These findings are discussed in relation to the 'vigilance avoidance' model of attention to threat stimuli. PMID- 12581687 TI - Attentional blink modulation in a reaction time task: performance feedback, warning stimulus modality, and task difficulty. AB - The present research investigated the effect of performance feedback on the modulation of the acoustic startle reflex in a Go/NoGo reaction time task. Experiment 1 (n = 120) crossed warning stimulus modality (acoustic, visual, and tactile) with the provision of feedback in a between subject design. Provision of performance feedback increased the number of errors committed and reduced reaction time, but did not affect blink modulation significantly. Attentional blink latency and magnitude modulation was larger during acoustic than during visual and larger during visual than during tactile warning stimuli. In comparison to control blinks, latency shortening was significant in all modality conditions whereas magnitude facilitation was not significant during tactile warning stimuli. Experiment 2 (n = 80) employed visual warning stimuli only and crossed the provision of feedback with task difficulty. Feedback and difficulty affected accuracy and reaction time. Whereas blink latency shortening was not affected, blink magnitude modulation was smallest in the Easy/No Feedback and the Difficult/Feedback conditions. PMID- 12581688 TI - Asymmetric interference in patterning discriminations: a case of modulated attention. AB - In sequential training positive patterning hinders negative patterning, but not vice versa. This asymmetry was attributed to interference of two different rules. In a Pavlovian skin conductance response conditioning experiment with humans, we further investigated this effect. Two groups performed sequential training of positive and negative patterning with a reduced number of trials per training schedule. In Group PNP, the sequence was positive patterning (A-, B-, AB+), negative patterning (C+, D+, CD-), followed by positive patterning (E-, F-, EF+). Group NPN consisted of negative patterning (A+, B+, AB-), positive patterning (C , D-, CD+), and negative patterning (E+, F+, EF-) again. As predicted by an attentional explanation we found no interference. Hence, the asymmetry observed earlier had been due to less attention to specific stimuli in consequence of an extensive use of the numerosity-rule. PMID- 12581689 TI - Sex role identity related to the ratio of second to fourth digit length in women. AB - Prenatal gonadal hormones have been implicated as important factors in the development of sex-role identity. The aim of the study reported here was to examine the relationship between adult sex-role preference and the second to fourth digit ratio (2D:4D ratio) in healthy women. There is evidence that the ratio of the length of second and fourth digits associates negatively with prenatal testosterone and positively with prenatal oestrogen. In this study the 2D:4D ratio was measured on a sample of 46 female university students. The subjects completed the form of the Bem Sex Role Inventory (BSRI). It was found that the lower 2D:4D ratios associated significantly with higher, masculinized bias scores in BSRI indicating that 2D:4D ratio predicts the female or male self reported sex-role identity in females. PMID- 12581692 TI - Pregnancy and the immune system: between tolerance and rejection. AB - Interactions between the conceptus and the mother are bi-directional: the feto placental tissues need nutrition and a suitable environment in homeostatic condition whereas the mother influenced by the placental factors adapts her metabolism and immune system. Many different mechanisms acting locally or at distance ensure tolerance of the semi-allogeneic graft by the maternal natural and adaptive immune defences. In front of this tolerance, mechanisms exist ensuring rejection of the conceptus by the mother (spontaneous abortion) through rupture of one or more tolerance mechanisms, notably in stress situations endangering the mother. Thus outcome of a pregnancy is dependent on efficiently working tolerance mechanisms, and rupture of such mechanisms can lead to rejection. The balance of influence leading either to tolerance or rejection is under control of internal (maternal and fetal) and external (environmental) factors. Rejection, if triggered, mainly occurs through immune-induced inflammation, tissue degradation and coagulation. PMID- 12581690 TI - The effects of pictures of emotional faces on tonic and phasic autonomic cardiac control in women and men. AB - The aim of the present study was to examine autonomic function in response to negatively and positively valenced pictures under different levels of conscious recognition. Heart period variability (HPV) and heart rate (HR) reactivity were studied in 53 males and females who were being shown pictures of angry and happy faces. The pictures, which were backwardly masked, were presented once every 30 s during a 5-min period and under three conditions (counterbalanced for type of facial expression): below the level of conscious recognition (17 ms), at an intermediate level (56 ms), and at a clearly recognizable level (2370 ms). Analyses of HR power spectrum (for 5 min in each condition) in the high frequency region (HF: 0.15-0.5 Hz) that reflects respiratory sinus arrhythmia, as well as analysis of phasic heart rate responses (7.5 s in 0.5 epochs following every picture presentation) were carried out. The main findings were that HF-power was higher, and cardiac midinterval acceleration lower, in response to angry as opposed to happy faces, a result obtained only for the men, however. No interaction effect between facial expression and the three exposure conditions was found, suggesting that the pictures induced emotional activation both subliminally and supraliminally. The results were discussed in terms of increased attention to aversive stimuli. PMID- 12581693 TI - Immunotoxicological consequences of perinatal chemical exposures: a plea for inclusion of immune parameters in reproduction studies. AB - The developing immune system may be especially vulnerable to the toxic effects of immunotoxicants. TCDD, a notorious immunotoxicant, has been shown to produce such effects in rodents as well as in man, and may be considered a prototype developmental immunotoxicant. A number of other chemicals have been identified that have been shown to affect the developing immune system. Yet, so far there are no immune parameters included in current OECD guidelines for developmental or reproductive toxicity testing. It is recommended that such tests are amended so that also the developing immune system is considered as a potential target of toxicity during developmental stages. PMID- 12581694 TI - Developmental immunotoxicity testing: a review. AB - There are currently no validated or widely accepted methods for evaluating the effects of a chemical on the developing immune system. Nonetheless because of concerns over children's health issues, specifically the possibility that the very young are uniquely susceptible to chemical perturbation, governmental regulators are beginning to ask for information about potential effects on the developing immune system. This paper will address the following three goals. First, an update of the regulatory pressures for developmental immunotoxicology will be presented from a U.S. perspective. Second, an update on the state of the science of developmental immunotoxicology will be presented with an emphasis on results from a recent collaboration between Dow, DuPont and Cornell University. Finally, the principle conclusions from a recent ILSI/HESI workshop will be summarized. At this workshop, it was generally acknowledged that there are a variety of techniques available for assessing immunosuppression in adult animal models. However, it was emphasized that there is uncertainty about how to apply these approaches to a developing animal, especially if the goal is to have some standard procedure(s) that could be applied for regulatory risk assessment. Ultimately, the primary conclusion from this workshop was that developmental immunotoxicology, as a science, is still in its infancy and is not ready to be applied in a risk assessment strategy. PMID- 12581695 TI - Reproductive toxicology studies and immunotherapeutics. AB - There are many ways in which immunotherapeutic agents could potentially cause developmental toxicity. According to the published data in humans and animals, however, this class of drugs does not appear to present any increased risk of reproductive toxicity by comparison with other therapeutic classes. The basic testing strategy outlined in the ICH guidelines is suitable for the preclinical reproductive toxicity assessment of these drugs (except vaccines). Particular consideration needs to be given to the choice of species when testing antibodies or human molecules. Immune-active drugs may have the potential to interfere with the development of the immune system. For this reason, it may be advisable to assess the integrity of the immune system in animals previously exposed to the test substance during development. Such tests can easily be appended to the study design of the pre- and post-natal study. In view of the lack of any provision for post-weaning exposure in the guidelines and the late maturation of the immune system, a separate post-natal study may be considered for drugs that are likely to be prescribed to children. PMID- 12581696 TI - Reproductive toxicity testing of vaccines. AB - Vaccines play a major role in the prevention of human birth defects by protecting the pregnant woman from teratogenic or otherwise harmful infections. Until now, it has not been common practice to perform preclinical developmental toxicity tests for new vaccines. Despite the excellent safety record of vaccines, increased attention is now being given to the feasibility of screening new vaccines for developmental hazards in animals before their use in humans. Contrary to previous assumptions, many vaccines are now given to potentially pregnant women. Any new components of the vaccine formulation (adjuvants, excipients, stabilisers, preservatives, etc.) could also be tested for influences on development, although based on past experience the risks are limited by the very low dosages used. The conferred immunity following vaccination lasts for several years. Therefore, the developing conceptus may theoretically be exposed to the induced antibodies and/or sensitised T-cells, even if the pregnant woman was last vaccinated during childhood (particularly if she encounters the antigen during pregnancy through exposure to infection). However, it should be kept in mind that viral or bacterial infections represent a higher risk for a pregnant woman than the potential adverse effects related to vaccination or the associated immune response. Non-clinical safety studies may be employed as an aid for hazard identification. In these studies interactions of the vaccine with the maternal immune system or with the developmental systems of the offspring are considered. Post-natal examinations are necessary to detect all possible manifestations of developmental toxicity, such as effects on the immune system. Species selection for the preclinical studies is based on immunogenicity to the vaccine and the relative timing and rate of transfer of maternal antibodies to the offspring. A single study design is proposed for the pre- and post-natal developmental assessments of vaccines in rodents and rabbits. PMID- 12581697 TI - Reproductive/developmental toxicity and immunotoxicity assessment in the nonhuman primate model. AB - Nonhuman primates are being used increasingly as a non-rodent animal model during preclinical toxicology and safety assessment on the basis of proven similarity and comparability between nonhuman primates and humans. The validity of the nonhuman primate models applies to many aspects of toxicological testing and holds particularly true for the evaluation of reproductive toxicology and developmental toxicology. More recently, the advent of humanized antibodies and vaccines imposed further demand on nonhuman primate models since many immunotherapeutics do not interact with rodent receptors but frequently only cross-react with primate tissue. In this paper we discuss the suitability of primate models for reproductive, developmental and immunotoxicology testing, and present our initial data on the development of lymphatic organs and immune system in a nonhuman primate model. PMID- 12581698 TI - Immunosuppressive drugs and cancer. AB - Among the many adverse effects induced by immunosuppressive drugs, cancers are a major cause of morbidity and mortality. This review is based on the most recent clinical data. Epidemiological studies and cancer registries have consistently shown an increased risk of malignancies in transplant patients although the calculated risk (4-500-fold increase) differs markedly between studies essentially because of differences in methodologies and selection of patients. Skin and lip cancers, lymphomas and Kaposi's sarcomas are the main types of cancer in these patients. A number of risk factors have been identified, such as latent viral infections, the treatment regimen and the level of immunosuppression. The increasing use of immunosuppressive drugs in nontransplant patients is useful to delineate more accurately the consequences of mild-to moderate immunosuppression. PMID- 12581699 TI - NK-cell activity in immunotoxicity drug evaluation. AB - NK-cell activity as a tool for detection of immunotoxic effects of new human drugs has gained further attention when the recent European note for guidance CPMP/SWP/1042/99 was adopted. The inclusion of NK-cell activity plus distribution of lymphocyte subsets were suggested as an alternative to the primary antibody response to a T-cell dependent antigen. Either of the two test alternatives should be included as a routine parameter in at least one repeated dose-toxicity study, rats or mice being the species of choice. The standard procedure for measuring NK-cell activity is the 51Cr-release assay. However, a new flow cytometric assay, adapted for rat peripheral blood, does not require dedicated groups of animals, offers the possibility of repeated testing, and shows at least as sensitive as the conventional 51Cr-release assay. PMID- 12581700 TI - Jewels among the junk. PMID- 12581701 TI - Encapsulated cell technology could prevent blindness. PMID- 12581702 TI - Asia bioinformatics: a new import-export industry. PMID- 12581705 TI - Products of yesteryear--bile beans. PMID- 12581707 TI - The use of combi chem, high-speed analog chemistry and HTS in drug discovery. PMID- 12581706 TI - Synthetic polymers in 21st century therapeutics: the way forward. PMID- 12581708 TI - Chemical genomics versus orthodox drug development. PMID- 12581709 TI - Smart high-throughput screening. PMID- 12581711 TI - Fulfilling the promise: drug discovery in the post-genomic era. AB - The genomic era has brought with it a basic change in experimentation, enabling researchers to look more comprehensively at biological systems. The sequencing of the human genome coupled with advances in automation and parallelization technologies have afforded a fundamental transformation in the drug target discovery paradigm, towards systematic whole genome and proteome analyses. In conjunction with novel proteomic techniques, genome-wide annotation of function in cellular models is possible. Overlaying data derived from whole genome sequence, expression and functional analysis will facilitate the identification of causal genes in disease and significantly streamline the target validation process. Moreover, several parallel technological advances in small molecule screening have resulted in the development of expeditious and powerful platforms for elucidating inhibitors of protein or pathway function. Conversely, high throughput and automated systems are currently being used to identify targets of orphan small molecules. The consolidation of these emerging functional genomics and drug discovery technologies promises to reap the fruits of the genomic revolution. PMID- 12581710 TI - A novel approach to identifying antiarrhythmic drug targets. AB - Sudden cardiac death, secondary to ventricular fibrillation (VF), remains the leading cause of death in the USA. Recent experimental and theoretical studies suggest that VF could be caused by spiral wave re-entry. The initiation and subsequent break-up of spiral waves has been linked to electrical alternans, a phenomenon produced in cardiac tissue that has a steeply sloped restitution relation. Agents that reduce the slope of the restitution relation have been shown to suppress alternans and, presumably by that mechanism, terminate VF. These results suggest that electrical restitution could be a promising new target for antiarrhythmic therapies. PMID- 12581712 TI - Advancing drug discovery through systems biology. AB - Pharmaceutical companies are facing an urgent need to both increase their lead compound and clinical candidate portfolios and satisfy market demands for continued innovation and revenue growth. Here, we outline an emerging approach that attempts to facilitate and alleviate many of the current drug discovery issues and problems. This is, in part, achieved through the systematic integration of technologies, which results in a superior output of data and information, thereby enhancing our understanding of biological function, chemico biological interactions and, ultimately, drug discovery. Systems biology is one new discipline that is positioned to significantly impact this process. PMID- 12581714 TI - Characterization and comparison of midgut proteases of Bacillus thuringiensis susceptible and resistant diamondback moth (Plutellidae: Lepidoptera). AB - The midgut proteases of the Bacillus thuringiensis resistant and susceptible populations of the diamondback moth, Plutella xylostella L. were characterized by using protease specific substrates and inhibitors. The midgut contained trypsin like proteases of molecular weights of 97, 32, 29.5, 27.5, and 25 kDa. Of these five proteases, 29.5 kDa trypsin-like protease was the most predominant in activation of protoxins of Cry1Aa and Cry1Ab. The activation of Cry1Ab protoxin by midgut protease was fast (T(1/2) of 23-24 min) even at a protoxin:protease ratio of 250:1. The protoxin activation appeared to be multi-step process, and at least seven intermediates were observed before formation of a stable toxin of about 57.4 kDa from protoxin of about 133 kDa. Activation of Cry1Aa was faster than that of Cry1Ab on incubation of protoxins with midgut proteases and bovine trypsin. The protoxin and toxin forms of Cry proteins did not differ in toxicity towards larvae of P. xylostella. The differences in susceptibility of two populations to B. thuringiensis Cry1Ab were not due to midgut proteolytic activity. Further, the proteolytic patterns of Cry1A protoxins were similar in the resistant as well as susceptible populations of P. xylostella. PMID- 12581715 TI - Attachment and germination of Entomophaga maimaiga conidia on host and non-host larval cuticle. AB - The lepidopteran-specific fungal pathogen Entomophaga maimaiga is highly virulent against Lymantria dispar (gypsy moth) larvae, and other members of the family Lymantriidae. Numerous species in the subfamily Cuculliinae (Family Noctuidae) are not susceptible to E. maimaiga due to the inability of this fungus to penetrate the larval cuticle. Conidial attachment and germination were compared among five cuculliine species and L. dispar using bioassays and scanning electron microscopy. Although conidia were showered evenly across larvae during bioassays, on L. dispar conidia were most abundant on segments, where they adhered well to the cuticle and germinated at high percentages. Conidia on cuculliine cuticles were predominantly found in large, loose aggregations in intersegmental areas. Few conidia on cuculliine cuticle germinated and scanning electron microscopy revealed a thick film of mucous enveloping conidia. We hypothesize that the conidia on cuculliines become coated by this film and were only loosely attached to the larval cuticle. No such film was seen on L. dispar larvae where individual conidia appeared well attached. On L. dispar larvae many conidia also adhered to setae. To determine if hydrophobicity affected the ability of E. maimaiga conidia to attach and germinate on a substrate, a goniometer was used to determine relative hydrophobicity of larval cuticles. L. dispar cuticle was more hydrophobic than cuculliine cuticle, suggesting that a high level of hydrophobicity could be a required characteristic for hosts. Cuticles from four cuculliine species and L. dispar were sequentially extracted using hexane, chloroform, and methanol. Conidia were showered onto glass slides coated with the different extracts and germination was quantified. Methanol extracts of cuculliine cuticle consistently decreased germination, compared to all extracts of L. dispar cuticle. For all L. dispar extracts, the majority of conidia produced germ tubes, which is a normal prerequisite for cuticular penetration. For the cuculliines, conidia exposed to hexane and chloroform extracts produced secondary conidia as did all controls, but the conidia exposed to cuculliine methanol extracts that germinated produced germ tubes. These studies demonstrated that a range of factors act in concert to prevent E. maimaiga infection of the cuculliine species investigated. PMID- 12581717 TI - Disintegration of the peritrophic membrane of silkworm larvae due to spindles of an entomopoxvirus. AB - Mode of action by which entomopoxvirus (EPV) spindles enhance nucleopolyhedrovirus (NPV) infection remains unclear. Spindles of Anomala cuprea entomopoxvirus (AcEPV), a coleoptran EPV, are known to enhance Bombyx mori NPV (BmNPV) infection in silkworm larvae. AcEPV spindles were orally administered to silkworm larvae with or without BmNPV polyhedra, and the peritrophic membranes (PMs) were observed using a binocular microscope. Soon after the larvae's access to spindles with or without the polyhedra had been terminated, some PMs disappeared wholly and some were observed in partial form. Some of the partial PMs observed were very fragile. The disintegration of the PM due to spindles also was observed by the histological sectioning of the midgut. However, a day after the larvae had terminated their access to the spindles, the PM regenerated partially or wholly. In contrast, the administration of AcEPV spheroids caused neither the disintegration of PMs nor the enhancement of BmNPV infection in silkworm larvae. These findings strongly suggest that the enhancement of NPV infection occurs due to that a greater number of NPV virions reaching the microvilli of midgut susceptible to NPV, since spindles lead to the disintegration of the PM as a barrier against NPV virions. PMID- 12581716 TI - Response of Penaeus indicus females at two different stages of ovarian development to a lethal infection with Vibrio penaeicida. AB - An association between vitellogenesis and the immune system was suggested in crustaceans from studies on plasma lipoproteins. The present research studies the effect of an experimentally induced bacterial infection on vitellogenesis in females of the shrimp Penaeus indicus, as a model for penaeid species. Pre vitellogenic and vitellogenic P. indicus females were experimentally infected with an extremely pathogenic bacterium, Vibrio penaeicida. The peak in mortality occurred earlier in pre-vitellogenic animals than in vitellogenic ones, although the final mortality level ( approximately 64-74%) 52h post-infection was nearly the same for the two groups. Twenty hours after infection, the total number of haemocytes was significantly reduced in vitellogenic females while there was no change in the pre-vitellogenic group. Protein synthesis in ovaries was not significantly affected by infection, at the two stages of ovarian development. No differences were found in mRNA levels of shrimp ovarian peritrophin protein (SOP), but preliminary results showed that mRNA expression of vitellin (VT) was reduced in a heavily infected vitellogenic female. The total amount of lipids in the haemolymph of vitellogenic females was almost twice higher than that of pre vitellogenic ones. However, there was no change in the total content of lipids, lipid classes and fatty acid distribution in haemolymph or hepatopancreas following infection. Although vitellogenic and pre-vitellogenic females probably respond differently to a lethal bacterial infection, physiological differences may be concealed by the rapid onset of mortality. PMID- 12581718 TI - Interaction of Leptomonas wallacei with the intestinal tract of its natural host Oncopeltus fasciatus (Hemiptera: Lygaeidae). AB - While investigating the distribution of Leptomonas wallacei in the intestine of the insect host Oncopeltus fasciatus, promastigotes and cyst-like forms of L. wallacei were observed only in the midgut ventricles V(3) and V(4) and the hindgut. In video-microscopy, once contact had occurred, the parasites remained attached to the midgut epithelium. Scanning electron microscopy revealed the adhesion of flagellates and cyst-like forms to the midgut wall and to the rectal pads of the hindgut. Using transmission electron microscopy, we observed that adhesion occurred mainly between the flagellum and the perimicrovillar membranes secreted by the midgut epithelium. No modifications were observed either in the parasite or in the epithelial cells. In the hindgut, adhesion to the superficial wax layer of the epithelial cells of the rectal pads was via flagellum. Host cell morphology appeared unaffected by L. wallacei. PMID- 12581719 TI - Improving baculovirus resistance to UV inactivation: increased virulence resulting from expression of a DNA repair enzyme. AB - The use of baculoviruses as biological control agents is hampered by their susceptibility to inactivation by ultraviolet (UV) light. In an attempt to reduce UV inactivation, an algal virus pyrimidine dimer-specific glycosylase, cv-PDG, was expressed in the baculovirus Autographa californica M nucleopolyhedrovirus (AcMNPV), and the infectivity of recombinant viruses expressing cv-PDG was measured after exposure to UV light. Expression of cv-PDG resulted in a 3-fold decrease in inactivation of budded virus by UV as measured by plaque assay in Spodoptera frugiperda Sf21 cells. However, occluded viruses expressing cv-PDG were not more resistant to UV inactivation than wild type AcMNPV when fed to either S. frugiperda or Trichoplusia ni neonate larvae. Surprisingly, however, viruses expressing cv-PDG showed a significant decrease in both the dose of occluded virus required for oral lethality and the time required for lethality compared to control virus, but these effects were only seen in S. frugiperda and not in T. ni larvae. PMID- 12581720 TI - Correlation between serovars of Bacillus thuringiensis and type I beta-exotoxin production. AB - beta-Exotoxin is a thermostable metabolite produced by some strains of Bacillus thuringiensis. Because of vertebrate toxicity, most commercial preparations of B. thuringiensis are prepared from isolates that do not produce beta-exotoxin. The aim of the present study was to find out the possible relationship between serovars of B. thuringiensis and beta-exotoxin production. A specific HPLC assay for type I beta-exotoxin has been used to detect this exotoxin in supernatants from final whole cultures of 100 strains belonging to four serovars of B. thuringiensis: thuringiensis, kurstaki, aizawai, and morrisoni. For each serovar, 25 strains randomly chosen from two Spanish collections were analyzed. Frequency of beta-exotoxin production was higher in B. thuringiensis serovar thuringiensis, whereas only two strains from serovar kurstaki showed beta-exotoxin production. None of the 25 strains belonging to serovars aizawai and morrisoni was found to produce this compound. Along with data from other studies, serovars can be classified as "common," "seldom," or "rare" beta-exotoxin producers. The serovar dependent beta-exotoxin production is discussed in relation to the evolutionary process of serovar differentiation, the plasmid compatibility and limited plasmid exchange between serovars, and with the serovar-dependent regulation of plasmid encoded genes. PMID- 12581721 TI - Characterization of cry1, cry2, and cry9 genes in Bacillus thuringiensis isolates from China. AB - Bacillus thuringiensis isolates from different ecological regions and sources of China were analyzed to study the distribution and diversity of cry genes and to detect the presence of novel cry genes. Strains containing cry1-type genes were the most abundant and represent 237 of the 310 B. thuringiensis isolates (76.5%). About 70 and 15.5% of the isolates contained a cry2 gene or cry9 gene, respectively, while 10.0% of the strains did not contain a cry1, cry2, or cry9 gene. Among the cry1 containing isolates, cry1A (67.7%), cry1I (60.6%), cry1C (43.9%), and cry1D (39.4%) genes were the most abundant. Forty-three different cry1 gene profiles were detected in this collection. Several cry1 genes were associated at a high frequency, such as the cry1C-cry1D and cry1A-cry1I gene combination. The cry1A and cry2 amplicons were digested with selected restriction enzymes to examine sequence diversity. Based on this RFLP analysis, one novel cry1A-type gene was observed. PMID- 12581722 TI - Survival of earthworms exposed to the slug-parasitic nematode Phasmarhabditis hermaphrodita. PMID- 12581723 TI - Hopelessness and risk behaviour among adolescents living in high-poverty inner city neighbourhoods. AB - Ethnographic literature on inner-city life argues that adolescents react to their uncertain futures by abandoning hope, leading them to engage in high levels of risk behaviour. However, few quantitative studies demonstrate this relationship. This study tests this relationship using a survey of 2468 inner-city adolescents, asking them questions about hopelessness, violent and aggressive behaviour, substance use, sexual behaviour, and accidental injury. Nearly 50% of males and 25% of females had moderate or severe feelings of hopelessness. Moreover, hopelessness predicted of each of the risk behaviours considered. These results suggest that effective prevention and intervention programmes aimed at inner-city adolescents should target hopelessness by promoting skills that allow them to overcome the limitations of hopelessness. PMID- 12581724 TI - The relationship of gender, gender identity, and coping strategies in late adolescents. AB - The relationships among gender, gender identity, and coping in late adolescents were examined. One hundred sixty-nine late adolescents completed measures assessing their masculinity and femininity, as well as their use of coping strategies. Females endorsed greater use of emotion-focused coping strategies than males. Late adolescents who were high in masculinity endorsed higher levels of problem-focused coping strategies than those who were low in masculinity. In contrast, late adolescents who were high in femininity endorsed higher levels of emotion-focused coping strategies than those who were low in femininity. Neither the gender nor the masculinity and femininity of late adolescents were predictive of the use of avoidant coping strategies. Overall, gender identity made an important and independent contribution to the endorsement of coping strategy use. These results emphasize the importance of assessing both gender and gender identity with regard to coping in late adolescents. PMID- 12581725 TI - Helpless attributions and depression in adolescents: the roles of anxiety, event valence, and demographics. AB - The relationship between helpless attributions and depression is well established. This study evaluated whether anxiety, event valence (positive or negative) and demographic variables (gender, age, socioeconomic status or race) influence this relationship. Results showed: (1) adolescents with anxiety and depression who were from lower socioeconomic backgrounds made less helpless attributions for negative events than did adolescents from higher socioeconomic backgrounds, (2) male adolescents with anxiety-only had helpless attribution styles that were similar to male adolescents with depression, but the same was not true for female adolescents, and (3) African-American adolescents showed less helpless attributions for negative events than did Caucasian adolescents. Results replicate past research showing that a helpless attribution style is associated with depression in adolescence, but suggest that this pattern may be influenced by demographic factors. Results also suggest that helpless attributions may be related to both anxiety and depression in males. PMID- 12581726 TI - Primary Mental Health Workers within Youth Offending Teams: a new service model. AB - Primary Mental Health Workers (PMHWs) have been deployed to address the mental health needs of young offenders referred to Youth Offending Teams (YOTs) in two UK areas. The mental health characteristics of 60 young people consecutively referred to these PMHWs, the assessment outcome and interventions offered, are described. In addition to the anticipated concerns about oppositional/aggressive behaviour, young people were referred for a range of mental health problems. There were high levels of emotional problems, self-harm, peer and family relationships difficulties, and school non-attendance. PMHWs offered a range of direct interventions, as well as consultation to YOT staff. The service findings indicate the usefulness of such an inter-agency model in strengthening the links between specialist CAMHS and YOTs, and providing an accessible, responsive and effective service to a needy group of young people. PMID- 12581727 TI - Shelters for houseless youth: a follow-up evaluation. AB - This study followed 345 Israeli youngsters who had been residents of two shelters for runaway and homeless youths, 6-12 weeks after their departure. Telephone interviews were conducted with the youngsters, their parents, and social workers in the community. A majority of the youngsters had either returned to their family homes, or had been placed out of home. Their residential stability was found to be low. Post-shelter place of residence was related to length of stay at the shelter, amount of contact with their family while at the shelter, and manner of departure. Our findings lead to a typology of shelter uses, and also raise questions about the extent to which shelters achieve their declared goals. PMID- 12581728 TI - Intergenerational value discrepancies in immigrant and host-national families and their impact on psychological adaptation. AB - This study investigated the relationship between intergenerational value discrepancies and psychological adaptation among adolescents with immigrant background (mean age=15.5 years) in Norway and Sweden. Results from two cohorts (parents and their children) of immigrant and host-national families in the two countries (n=574) indicated that immigrant adolescents on the whole neither differed from their host peers with respect to psychological adaptation nor on value discrepancies. Value discrepancies were weakly related to psychological adaptation. Results are discussed as to whether the findings may reflect a general phenomenon for all adolescents or specific to those with immigrant background. PMID- 12581729 TI - Visibility management in school and beyond: a qualitative study of gay, lesbian, bisexual youth. AB - Grounded theory was utilized to develop a theory of gay, lesbian, and bisexual (GLB) youths' interactions with and response to their environment. Data included in-depth interviews of 20 GLB youth. The central phenomenon that emerged from the qualitative data analysis was the strategy of visibility management. Visibility management refers to the dynamic, ongoing process by which GLB youth make careful, planned decisions about whether they will disclose their sexual orientation, and, if they decide to disclose, to whom and how they disclose, and how they continue to monitor the presentation of their sexual orientation in different environments. The process of visibility management is proposed as a key construct in GLB identity development. Implications are proposed for working with GLB youth and their families and for making schools safer for GLB youth. PMID- 12581730 TI - Training general practitioners in the identification and management of adolescent depression within the consultation: a feasibility study. AB - BACKGROUND: Depressive disorders are common in adolescent general practice attenders. METHOD: Adolescent attenders were screened/interviewed for depressive disorders, general practitioners (GPs) completed a checklist indicating recognition of psychopathology prior to and following GP training in the identification/management of adolescent depression. RESULTS: One hundred and thirty consecutive adolescent attenders were screened before and 184 after training. Ten GPs completed the training. Psychiatric interviews with 38 adolescents with high depressive scores prior to and 44 following training identified 10 (26%) and 21 (48%), respectively, as clinically depressed. Sensitivity of GP identification improved from 2/10 (20%) to 9/21 (43%) without loss of specificity; predictive validity from 2/6 (33%) to 9/12 (75%). Adolescents interviewed appreciated the intervention. CONCLUSIONS: Training GPs is feasible and may improve recognition of adolescent depression. PMID- 12581733 TI - Quantification of DNA adducts in individual cells by immunofluorescence: effects of variation in DNA conformation. AB - Previously reported detection of melphalan-DNA adducts by immunofluorescent staining indicated considerable intercell variation in fluorescence levels. Investigations were undertaken to determine whether this variation reflected actual intercell differences in adduct levels. Melphalan-treated CCRF-CEM leukaemia cells were analysed by the trapped-in-agarose DNA immunostaining (TARDIS) method using fluorescein immunofluorescence and Hoechst dye-DNA fluorescence. Increasing the time of DNA denaturation in alkali affected the staining intensity, in agreement with known adduct properties, but failed to reduce intercell heterogeneity. To test the hypothesis that heterogeneity resulted from variation in levels of DNA strand breaks, drug-treated cells were exposed to ionising radiation. An increase in level and reduction in heterogeneity of immunofluorescence were observed, optimal at 10 Gy. When samples were irradiated after lysis, 1 Gy was optimal. At the optimal doses, irradiation before or after lysis resulted in similar levels of DNA strand breaks. Our conclusions are as follows: (a) There was no major intercell variation in the number of adducts other than from variation in DNA content. (b) Detection of melphalan, and possibly other adducts, by immunofluorescence can be markedly influenced by the level of strand breaks present in the DNA. (c) Samples analysed for melphalan adducts by immunofluorescence should be irradiated to minimise errors due to this factor. PMID- 12581734 TI - Staurosporine-induced death of MCF-7 human breast cancer cells: a distinction between caspase-3-dependent steps of apoptosis and the critical lethal lesions. AB - To test the role of caspase 3 in apoptosis and in overall cell lethality caused by the protein kinase inhibitor staurosporine, we compared the responses of MCF 7c3 cells that express a stably transfected CASP-3 gene to parental MCF-7:WS8 cells transfected with vector alone and lacking procaspase-3 (MCF-7v). Cells were exposed to increasing doses (0.15-1 microM) of staurosporine for periods up to 19 h. Apoptosis was efficiently induced in MCF-7c3 cells, as demonstrated by cytochrome c release, processing of procaspase-3, procaspase-8, and Bid, increase in caspase-3-like DEVDase activity, cleavage of the enzyme poly(ADP-ribose) polymerase, DNA fragmentation, changes in nuclear morphology, and TUNEL assay and flow cytometry. For all of these measures except cytochrome c release, little or no activity was detected in MCF-7v cells, confirming that caspase-3 is essential for efficient induction of apoptosis by staurosporine, but not for mitochondrial steps that occur earlier in the pathway. MCF-7c3 cells were more sensitive to staurosporine than MCF-7v cells when assayed for loss of viability by reduction of a tetrazolium dye. However, the two cell lines were equally sensitive to killing by staurosporine when evaluated by a clonogenic assay. A similar distinction between apoptosis and loss of clonogenicity was observed for the cancer chemotherapeutic agent VP-16. These results support our previous conclusions with photodynamic therapy: (a) assessing overall reproductive death of cancer cells requires a proliferation-based assay, such as clonogenicity; and (b) the critical staurosporine-induced lethal event is independent of those mediated by caspase-3. PMID- 12581735 TI - Altered response of vascular smooth muscle cells to exogenous biochemical stimulation in two- and three-dimensional culture. AB - Removal of vascular smooth muscle cells (SMC) from their native environment alters the biochemical and mechanical signals responsible for maintaining normal cell function, causing a shift from a quiescent, contractile phenotype to a more proliferative, synthetic state. We examined the effect on SMC function of culture on two-dimensional (2D) substrates and in three-dimensional (3D) collagen Type I gels, including the effect of exogenous biochemical stimulation on gel compaction, cell proliferation, and expression of the contractile protein smooth muscle alpha-actin (SMA) in these systems. Embedding of SMC in 3D collagen matrices caused a marked decrease in both cell proliferation and expression of SMA. The presence of the extracellular matrix modulated cellular responses to platelet-derived growth factor BB, heparin, transforming growth factor-beta1, and endothelial cell-conditioned medium. Cell proliferation and SMA expression were shown to be inversely related, while gel compaction and SMA expression were not correlated. Taken together, these results show that SMC phenotype and function can be modulated using biochemical stimulation in vitro, but that the effects produced are dependent on the nature of the extracellular matrix. These findings have implications for the study of vascular biology in vitro, as well as for the development of engineered vascular tissues. PMID- 12581736 TI - ER stress induces caspase-8 activation, stimulating cytochrome c release and caspase-9 activation. AB - Excess ER stress induces caspase-12 activation and/or cytochrome c release, causing caspase-9 activation. Little is known about their relationship during ER stress-mediated cell death. Upon ER stress, P19 embryonal carcinoma (EC) cells showed activation of various caspases, including caspase-3, caspase-8, caspase-9, and caspase-12, and extensive DNA fragmentation. We examined the relationship between ER stress-mediated cytochrome c/caspase-9 and caspase-12 activation by using caspase-9- and caspase-8-deficient mouse embryonic fibroblasts and a P19 EC cell clone [P19-36/12 (-) cells] lacking expression of caspase-12. Caspase-9 and caspase-8 deficiency inhibited and delayed the onset of DNA fragmentation but did not inhibit caspase-12 processing induced by ER stress. P19-36/12 (-) cells underwent apoptosis upon ER stress, with cytochrome c release and caspase-8 and caspase-9 activation. The dominant negative form of FADD and z-VAD-fmk inhibited caspase-8, caspase-9, Bid processing, cytochrome c release, and DNA fragmentation induced by ER stress, suggesting that caspase-8 and caspase-9 are the main caspases involved in ER stress-mediated apoptosis of P19-36/12 (-) cells. Caspase 8 deficiency also inhibited the cytochrome c release induced by ER stress. Thus, in parallel with the caspase-12 activation, ER stress triggers caspase-8 activation, resulting in cytochrome c/caspase-9 activation via Bid processing. PMID- 12581737 TI - Analysis of the cellular heterogeneity in the basal layer of mouse ear epidermis: an approach from partial decomposition in vitro and retroviral cell marking in vivo. AB - In the thin epidermis, the existence of epidermal proliferation units was hypothesized. Each unit is supposed to be partitioned into each column of polygonal-shaped cornified plates, estimated to contain a central stem cell in its basal layer. We attempted to verify this hypothesis in vitro by analyzing the partially decomposed fragment of mouse ear epidermis and in vivo using retroviral cell marking. Partially decomposed fragments of the mouse ear epidermis, mostly composed of cytokeratin 14-expressing basal keratinocytes, formed multicellular colonies in vitro. They were composed of heterogeneously shaped cells, morphologically resembling the cells in each single cell-derived colony, including potential stem cells with great proliferative potency in vitro. The estimated frequency of the candidates of stem cells in the fragments was much lower than the prediction from the representative hypothesis. Retroviral cell marking with nuclear localizing LacZ protein in vivo suggested the existence of a large clonal cellular unit for epidermal renewal. From these in vitro and in vivo observations, we propose a new model for the epidermal proliferation unit. PMID- 12581738 TI - Proto-oncoprotein TLS/FUS is associated to the nuclear matrix and complexed with splicing factors PTB, SRm160, and SR proteins. AB - TLS/FUS is a nucleic acid-binding protein whose N-terminal half functions as a transcriptional activator domain in fusion oncoproteins found in human leukemias and liposarcomas. Previous reports have suggested a role for TLS/FUS in transcription and splicing processes. Here we report the association of TLS/FUS with the nuclear matrix and investigate its role in splicing. Splicing of two pre mRNAs was inhibited in a TLS/FUS-immunodepleted extract and could only be partly restored by addition of recombinant TLS/FUS or/and SR proteins, known interaction partners of TLS/FUS. The subsequent analysis of TLS/FUS immunoprecipitates revealed that, in addition to the SR proteins SC35 and SRp75, the splicing factor PTB (hnRNPI) and the splicing coactivator SRm160 are complexed with TLS/FUS, thus explaining the inability to restore splicing completely. Coimmunolocalization confirmed the nuclear matrix association and interaction of TLS/FUS with PTB, SR proteins, and SRm160. Our results suggest that the matrix protein TLS/FUS plays a role in spliceosome assembly. PMID- 12581739 TI - Mitotic activity of rat muscle satellite cells in response to serum stimulation: relation with cellular metabolism. AB - Cellular and molecular adaptations of satellite cells isolated from rat hindlimb muscles (n = 10) were investigated in response to serum stimulation. Flow cytometry analysis of the amounts of DNA and RNA indicated that 97.7 +/- 0.7% of satellite cells were in G0 at the end of the isolation procedure, whereas 93.2 +/ 2.0% of cells were cycling after serum exposure. The length of cell division was 34.0 +/- 2.8 h. Myoblast proliferation was asynchronous, suggesting the existence of heterogeneous cell populations in skeletal muscle. Myoblast proliferation was also accompanied by a significant increase in c-met expression, and major adaptations of energetic and proteolytic metabolisms, including an increase in the relative contribution of glycolytic metabolism for energy production, an increase in proteasome and matrix metalloproteinases 2 and 9 activities, and a decrease in plasminogen activator activities. Our data suggest that, along with molecular adaptations leading to cell cycle activation itself, adaptations in energetic and proteolytic metabolisms are crucial events involved in satellite cell activation and myoblast proliferation. PMID- 12581740 TI - Intracellular and extracellular cathepsin B facilitate invasion of MCF-10A neoT cells through reconstituted extracellular matrix in vitro. AB - Lysosomal cysteine proteinase cathepsin B is implicated in remodeling the extracellular matrix, a crucial step in the process of tumor cell invasion. In this study the contributions of intracellular and extracellular cathepsin B activities in the invasion of ras-transformed human breast epithelial cells, MCF 10A neoT, were assessed using specific cathepsin B neutralizing monoclonal antibody (Mab) 2A2, together with other general and specific cysteine proteinase inhibitors. We showed that the degradation of extracellular matrix by living MCF 10A neoT cells was predominantly intracellular, as imaged by confocal assays using quenched fluorescent substrate DQ-collagen IV. CA-074, a membrane impermeable cathepsin B-selective inhibitor and its membrane-permeable analogue CA-074Me showed similar inhibition of invasion at 10 microM, i.e., 24.9 and 27.0%, respectively. Neutralizing monoclonal antibody exhibited a significantly higher inhibitory effect, decreasing invasion at 0.5 microM by 42.7%. Tumor cells may internalize monoclonal antibody; therefore, 2A2 Mab could impair both the intracellular and the extracellular fractions of cathepsin B activity. However, both 2A2 Mab and cathepsin B-selective inhibitors were less potent than the general cysteine proteinase inhibitors chicken cystatin and E-64, indicating that other cysteine proteinases, presumably cathepsin L, are involved in invasion. Our results show that intracellular and extracellular cathepsin B activity contribute to in vitro invasion of MCF-10A neoT cells and suggest that inhibitors capable of impairing both fractions have a potential as new anticancer drugs. PMID- 12581741 TI - Keratinocyte growth factor promotes cell motility during alveolar epithelial repair in vitro. AB - Epithelia play a key role as protective barriers, and mechanisms of repair are crucial for restoring epithelial barrier integrity, especially in the lung. Cell spreading and migration are the first steps of reepithelialization. Keratinocyte growth factor (KGF) plays a key role in lung epithelial repair and protects against various injuries. We hypothesized that KGF may protect the lung not only by inducing proliferation but also by promoting epithelial repair via enhanced epithelial cell migration. In an in vitro wound-healing model, we found that KGF enhanced wound closure by 33%. KGF acted primarily by inducing lamellipodia emission (73.2 +/- 3.9% of KGF-treated cells had lamellipodia vs 61.3 +/- 3.4% of control cells) and increasing their relative surface area (59 +/- 2.7% with KGF vs 48 +/- 2.0% in controls). KGF reduced cytoskeleton stiffness as measured by magnetic twisting cytometry and increased cell motility (5.8 +/- 0.42 microm/h with KGF vs 3.7 +/- 0.41 microm/h in controls). KGF-increased cell motility was associated with increased fibronectin deposition during wound closure and with fibronectin reorganization into fibrils at the rear of the cells. Taken together, our findings strongly suggest that KGF may promote epithelial repair through several mechanisms involved in cell migration. PMID- 12581742 TI - Human endostatin-derived synthetic peptides possess potent antiangiogenic properties in vitro and in vivo. AB - Pharmacological control of the angiogenic process (i.e., the neovascularization necessary for the growth and progression of tumors and metastases) is considered to be one of the most promising approaches to antineoplastic therapy. Endostatin, a 20-kDa protein derived from collagen XVIII, is one of the first recently discovered endogeneous antiangiogenic substances, but its cell targets and mechanism(s) of action are still unknown. We thought it would be interesting to test whether shorter peptides derived from endostatin might preserve its antiangiogenic activity. Four synthetic peptides corresponding to the sequences 6 49 (I), 50-92 (II), 93-133 (III), and 134-178 (IV) of human endostatin were tested for their ability to inhibit endothelial cell proliferation, migration, and both in vitro and in vivo angiogenesis. Fragment I (and fragment IV in the tests performed) was found to be fully biologically active in all of the angiogenesis assays, and sometimes showed even greater potency and efficacy than full-length human endostatin itself. PMID- 12581743 TI - Negative regulation of the protection of eIF2alpha phosphorylation activity by a unique acidic domain present at the N-terminus of p67. AB - Eukaryotic initiation factor 2 (eIF2)-associated glycoprotein, p67, has protection of eIF2alpha phosphorylation (POEP) activity, and this activity requires lysine-rich domains I and II of p67. Another unique acidic residue-rich domain is also present at the N-terminus of p67. In this study we analyzed the role of this acidic residue-rich domain in POEP activity. Our data revealed that constitutive expression of a mutant form of p67 (D6/2) in mammalian cells resulted in increased POEP activity, and this activity was partially inhibited when second-site alanine substitutions at the conserved amino acids D251, D262, E364, and E459 were introduced in the D6/2 mutant. In contrast, a similar mutation at the conserved H331 position did not show any effect on POEP activity. Individual alanine substitutions at the above conserved amino acids in wild-type p67 did not show any significant effect on POEP activity except the E459 position where alanine substitution caused approximately 50% increase in POEP activity as compared to the wild type. Although, the levels of endogenous p67 and p67 deglycosylase did not correlate with the POEP activity, we found that the D6/2 mutant of p67 was glycosylated at a higher level in mammalian cells as compared to wild-type p67. The increased POEP activity of the D6/2 mutant also correlated with the higher rate of overall protein synthesis in mammalian cells constitutively expressing this mutant form of p67. Taken together, these data suggest that the acidic residue-rich domain present at the N-terminus of p67 may have a negative role in POEP activity. PMID- 12581744 TI - Inhibition of breast carcinoma and trophoblast cell invasiveness by vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen and angiogenic growth factor that enhances endothelial cell invasion through the extracellular matrix (ECM). While various cell types express VEGF receptors, little is known about the biological actions of VEGF on nonendothelial cells. Therefore, the main objective of the present study was to determine the effect of VEGF on the in vitro invasiveness and proliferation of human MDA-MB-231 breast carcinoma cells and human HTR-8/SVneo trophoblast cells. Reverse-transcriptase polymerase chain reaction analysis demonstrated the presence of transcripts encoding VEGF receptors (VEGFR) -1, -2, and -3 as well as neuropilins-1 and -2 in the trophoblast cells, and the presence of transcripts encoding VEGFR-2 and neuropilins-1 and -2 in the breast carcinoma cells. Both cell lines also expressed transcripts for VEGF-A, -B, -C and -D, as well as for placenta growth factor (PlGF). Although incubation with exogenous VEGF-A(165) or VEGF-A(121) did not affect the rate of proliferation of either the trophoblast or the breast carcinoma cells, incubation with these molecules reduced their ability to invade through reconstituted ECM (Matrigel). The effect of VEGF-A(165) on the invasiveness of both cell lines was inhibited by the inclusion of a neutralizing antibody to VEGF. Exogenous VEGF-A(165) also decreased the cell surface expression of the urokinase-type plasminogen activator (a molecule required for invasion) by the breast carcinoma and trophoblast cells. These results indicate that the biological actions of VEGF on certain cell types may differ from the effects of this molecule on vascular endothelial cells, and therefore are relevant to angiogenesis-based therapies. PMID- 12581745 TI - Evidence for cytoskeletal changes secondary to plasma membrane functional alterations in the in vitro cell response to Clostridium perfringens epsilon toxin. AB - To investigate the mode of action of Clostridium perfringens epsilon-toxin, MDCK cells were treated with purified toxin and incubated at 37 degrees C for up to 24h. Exposure to epsilon-toxin caused a time-dependent decrease in cell-cell and cell-substrate interactions. After 30min of treatment retraction of the cell body and the emission of filopodia were detectable in a number of cells. Longer exposure resulted in cell rounding and cell blebbing which reached a maximum after 5h of toxin treatment. A parallel modification in the cytoskeleton was also detected. Actin marginalization and the entanglement of microtubules and intermediate filaments were observed by fluorescence microscopy after 30min of toxin exposure. Functional alterations of the plasma membrane of MDCK cells were assessed by flow cytometry. After 10 or 30min of intoxication an increase in cell volume was detected, indicating an alteration in plasma membrane permeability. These findings provide evidence for cytoskeletal changes and plasma membrane functional alterations in the in vitro cell response to C. perfringens epsilon toxin. PMID- 12581746 TI - In vitro study of Listeria monocytogenes infection to murine primary and human transformed B cells. AB - Immunity to Listeria monocytogenes is largely mediated by T lymphocytes. Recently, B lymphocytes or their secreted products are implicated to provide immunity against L. monocytogenes infection. To understand whether L. monocytogenes can infect and kill B cells as a possible strategy to initiate an infection, we examined the effects of L. monocytogenes on a human B lymphoma (Ramos RA-1) and mouse primary B cells in vitro. L. monocytogenes infection resulted in significantly (p0.5 mm) intimal thickening experience cardiac events. We sought to contrast clinical characteristics of heart transplant recipients who have prognostically relevant, severe intimal proliferation and who experience cardiac events with those who remain event free. METHODS: We prospectively followed an inception cohort of 54 consecutive heart transplant recipients with severe intimal proliferation (intimal thickness >0.5mm) of the coronary arteries after index intravascular ultrasound examination to assess the development of cardiac events (sudden cardiac death, myocardial infarction) and/or the necessity for coronary revascularization with percutaneous techniques (angioplasty, atherectomy, stent implantation) or surgical bypass. RESULTS: Based on the occurrence of adverse cardiac events during the subsequent 24 months, we divided the study cohort into 2 groups: Group 1 (no event, n = 33) and Group 2 (cardiac event, n = 21). Both groups demonstrated similar intimal thickness at the index ultrasound (Group 1, 0.89 +/- 0.27 mm, vs Group 2, 0.94 +/- 0.36 mm; p = not significant). Those with cardiac events were more likely than those without events to have hyperlipidemia, to have greater exposure to cumulative and average daily prednisone, and to exhibit greater average biopsy rejection scores at follow-up. CONCLUSIONS: These observations underscore the importance of the quality and not merely the quantity of intimal proliferation in determining occurrence of morbid cardiac events and further emphasize the interaction of immunologic and non-immunologic factors in determining event vulnerability in cardiac allograft vasculopathy. PMID- 12581757 TI - Proposition: the benefit of cardiac transplantation in stable outpatients with heart failure should be tested in a randomized trial. AB - Recent data suggest that cardiac transplantation is associated with a survival benefit only in patients at high risk for dying of advanced heart failure without this procedure. To test the hypothesis that survival and quality of life advantages associated with cardiac transplantation exist in stable outpatients, a 3-stage approach is proposed: 1). to establish a database within the International Society for Heart and Lung Transplantation/United Network for Organ Sharing/Eurotransplant infrastructure that will provide an estimate of the survival benefit of heart transplantation in various heart failure risk strata by prospectively following cohorts of patients listed for heart transplantation; 2). to organize an international consensus conference that will define, based on the review of the Stage 1 data, the feasibility of a prospective randomized trial; and 3). pending consensus, to perform a clinical trial, perhaps with an augmented, randomized design that allocates cardiac transplantation to all patients at high risk for dying of heart failure while randomizing patients at low risk to either conventional treatment or cardiac transplantation. Generating such scientific evidence is important in light of today's donor organ crisis and the associated difficulties of equitable resource allocation. PMID- 12581759 TI - Coronary atherosclerosis in cardiac transplant patients treated with total lymphoid irradiation. AB - BACKGROUND: Multiple episodes of rejection following cardiac transplantation have been associated with an increased incidence of coronary atherosclerosis. Total lymphoid irradiation (TLI) has been shown to be a successful treatment for persistent allograft rejection, but its effect on coronary arterial disease has yet to be evaluated. METHODS: From 1987 to 1999, 40 patients required TLI for persistent or recurrent allograft rejection following heart transplantation. Each patient's (Group 1, n = 31) post-transplant coronary angiograms were examined and compared with those of a control group (Group 2, (n = 32) matched for time of transplantation. Degree of coronary stenosis was assessed on a 6-point scale. All patients received induction therapy (rabbit anti-thymocyte globulin or OKT3) and standard triple immunosuppressive therapy. TLI (80 cGy x 10 fractions) was used for the treatment of recurrent or persistent rejection on the basis of clinical indications. Actuarial survival, number and treatment of rejection episodes, and severity of coronary artery disease were compared in each group. RESULTS: Recipient gender, age, race and cytomegalovirus (CMV) status at time of transplant, along with donor gender, CMV status and graft ischemia time, were similar in both groups. Group 1 donor age was younger than that of Group 2 (22.2 +/- 11.2 vs 31.5 +/- 13.6 years, p = 0.004), and the indication for surgery in Group 1 patients was more likely to be ischemic heart disease (15 of 31 vs 6 of 32, p = 0.02). Mean follow-up was 5.7 +/- 3.5 years in Group 1 vs 6.9 +/- 3.8 in Group 2 (p = NS). Group 1 had more rejection episodes (4.4 +/- 2.2 vs 2.3 +/- 2.0, p = 0.0002) and more steroid treatments (9.78 +/- 4.0 g vs 5.14 +/- 4.7 g, p < 0.0001), but less coronary artery disease compared with Group 2 (p = 0.035). CONCLUSIONS: Despite multiple episodes of rejection, patients treated with TLI after cardiac transplant appear to develop less coronary atherosclerosis than appropriately matched controls. PMID- 12581760 TI - Prognosis of moderate coronary artery lesions in heart transplant patients. AB - BACKGROUND: Despite recent advances in our understanding of allograft vasculopathy, little is known about the evolution of moderate coronary lesions in heart transplant recipients. METHODS: We retrospectively analyzed 58 heart transplant patients undergoing annual coronary angiography who demonstrated a moderate lesion (>30% and <60% diameter stenosis) on any routine annual study. In an attempt to find criteria that could distinguish such patients who were at high risk of disease progression from those at low risk, we reviewed the clinical and biologic features and angiographic and clinical outcomes of patients with and without lesion progression at 1 year. RESULTS: Of the 58 patients who had an initially moderate coronary lesion, 28 (48%) showed progression of the lesion at angiography 1 year later (occlusion of the culprit vessel or progression to a severe lesion >60%) that required revascularization (angioplasty or bypass surgery). The 30 remaining patients showed no lesion progression. At the time of the first angiogram the only criterion which could predict lesion progression at 1 year was the presence of multi-vessel disease (p < 0.0001). Prognosis for these patients was poorer than in those with no disease progression, with a higher proportion of revascularization and sudden death (p < 0.001). Patients without lesion progression at 1 year had neither clinical events nor significant subsequent lesion progression during a mean follow-up of 6 years. CONCLUSIONS: The presence of a moderate coronary stenosis in heart transplant patients justifies a repeat angiogram 1 year later. The use of percutaneous coronary angioplasty in such patients has not been validated, but may be an option to delay or prevent progression to coronary occlusion. PMID- 12581761 TI - Dobutamine stress echocardiography predicts myocardial improvement in patients supported by left ventricular assist devices (LVADs): hemodynamic and histologic evidence of improvement before LVAD explantation. AB - BACKGROUND: Cardiac function may improve in patients with end-stage heart failure who receive long-term support (>30 days) with left ventricular assist devices (LVADs). Dobutamine stress echocardiography (DSE) has been used to quantitate myocardial recovery in patients with heart failure supported with LVADs. By recording the hemodynamic response with the use of DSE, we evaluated and applied the resulting data to patients receiving LVAD support. METHODS AND RESULTS: The study population included 16 patients who underwent LVAD implantation, regained functional capacity on full LVAD support, and tolerated decreased mechanical support with no worsening of dyspnea or fatigue. All 16 patients underwent dobutamine stress with increasing doses of dobutamine (from 5 to 40 mcg/kg/min). Hemodynamics and 2-dimensional (2-D) echocardiography was performed at each dose level. In addition, paired myocardial samples were obtained and analyzed histologically to determine myocyte size and collagen content. Dobutamine stress separated the study population into 2 groups: those who had favorable responses to dobutamine (9/16) and those who had unfavorable responses (i.e., experienced hemodynamic deterioration; 7/16). Favorable dobutamine responses were characterized by improved cardiac index, improved force-frequency relationship in the left ventricle (dP/dt), improved left ventricular ejection fraction, and decreased left ventricular end-diastolic dimension. All 9 favorable responders underwent LVAD explantation, and 6 survived for more than 12 months. In all patients studied, LVAD support resulted in decreased myocyte size (n = 14, 33.9 +/- 0.9 microm before vs 16.6 +/- 0.8 microm after support, p = 0.0001; normal, 5 15 microm) but resulted in no consistent changes in collagen content. CONCLUSIONS: Dobutamine stress echocardiography with hemodynamic assessment may be a useful tool in assessing physiologic improvement in myocardial function of patients with end-stage heart failure who receive LVAD support. It may help predict which patients can tolerate LVAD removal. Prospective analysis of cardiac function is now warranted to better define myocardial recovery in patients supported with LVADs. PMID- 12581762 TI - Outcome of children with end-stage congenital heart disease waiting for cardiac transplantation. AB - BACKGROUND: End-stage congenital heart disease (CHD) is a major indication for pediatric cardiac transplantation. The objective of the study was to evaluate pre transplant outcome of children with CHD. METHODS: The clinical profile and outcome of patients with CHD <20 years of age listed for transplantation (1993 to 1999) were reviewed and patients who died waiting (Group I) were compared with survivors to transplant (Group II). RESULTS: Mean age of the patients (n = 46) was 8.3 +/- 8 years. Primary indications for transplant were ventricular dysfunction in 36 (78%), failed Fontan in 8 (18%) and severe hypoxemia in 2 (4%) patients. Thirty-two patients were Status 1 (70%), 14 were Status 2 and 5 patients were de-listed. Twenty-nine of the 41 patients that remained listed survived to transplant, 12 (29%) died waiting. Causes of death were sepsis in 2 and severe heart failure (HF) in 10 patients. Eight patients died with multi system organ failure, including 3 on mechanical circulatory support. Mean time to death was 29 +/- 28 days and time to transplant was 94 +/- 176 days. Mean age at listing was younger in Group I (2.6 +/- 4 years) compared with Group II (9.1 +/- 7 years, p < 0.05). Mean HF duration was shorter in Group I (3.6 +/- 3.9 months) compared with Group II (25 +/- 33 months, p < 0.05). Fifty-day actuarial survival on the waiting list was lower in infants (38%) compared with older children (91%, p < 0.05). In contrast to the high mortality (71%) in infants with CHD, all infants with cardiomyopathy survived to transplant. CONCLUSIONS: Seventy-one percent of patients listed with CHD survived to transplant. Younger age at listing and rapid onset of HF were significant risk factors for pre-transplant mortality. PMID- 12581763 TI - Lung transplantation in children following treatment for malignancy. AB - BACKGROUND: End-stage lung disease is a rare complication of treatment for hematologic and solid tumors in children. When present, it is generally progressive, resulting in the patient being cured of cancer only to die of respiratory failure. Lung transplantation is believed by some to be of overly high risk in this population because of the pre-existing malignancy as well as the presumed compromised immune status. METHODS: Six children (Group 1), aged 5 to 17 years (mean 12.4 years), underwent lung transplantation at our institution because of either pulmonary fibrosis or bronchiolitis obliterans following treatment for acute leukemia (n = 5) or medulloblastoma (n = 1). All patients received chemotherapy, radiation and bone marrow transplantation in the course of treatment for their malignancies. The average forced expiratory volume in 1 second (FEV(1)) was 16% of the predicted value and forced vital capacity (FVC) was 28% of predicted. These results were compared with a similar group of 13 patients undergoing lung transplantation at our institution during the same time interval (Group 2). RESULTS: There were 2 deaths in Group 1, 1 early and 1 late, for an overall survival of 67% at a mean follow-up of 4.02 years. There were no early and 7 late deaths in Group 2 for an overall survival of 46% at a mean follow-up of 4.8 years. The acute rejection rate in the first post-transplant year was 0.2 episode/patient in Group 1 and 1.8 episode/patient in Group 2 (p <0.01). No patient in Group 1 has developed post-transplant lymphoproliferative disease or a relapse of their primary malignancy. Two patients in Group 1 developed unusual infections-Aspergillus and Mycobacterium chelonae. No such infections occurred in Group 2. CONCLUSIONS: Although this represents a small series, we believe that patients with respiratory failure following treatment of a prior malignancy are suitable candidates for lung transplantation. Although they may have some relative protection from acute rejection episodes by virtue of an already compromised immunologic status while receiving standard immunosuppression, an increased propensity for opportunistic infection may exist. PMID- 12581764 TI - Indirect fluorescent antibody testing of nasopharyngeal swabs for influenza diagnosis in lung transplant recipients. AB - BACKGROUND: Rapid and reliable diagnosis of respiratory viral infections (RVI) in lung transplant recipients is essential to direct therapy of acute graft dysfunction and identify epidemic trends. Traditional techniques of serology and viral culture are limited by the lack of antibody response and delay in diagnosis. METHODS: We examined the clinical utility of indirect fluorescent antibody (IFA) testing in adult lung transplant patients with suspected RVI, compared with serology and culture. Nasopharyngeal and throat swabs (NT) were obtained to sample epithelial cells, followed by application of monoclonal antibody to respiratory syncytial virus, adenovirus, parainfluenza 1-3 and influenza A and B. The Bartels Respiratory Viral Detection kit was used with IFA results available within 24 hours. RESULTS: Nine of 18 patients tested positive for RVI with influenza A (n = 8) and influenza B (n = 1) detected. The sensitivity of IFA (67%) was higher than that of cell culture (45%). With intensive supportive therapy, infection was self-limiting in bronchiolitis obliterans syndrome (BOS) Grade 0-2 patients. However, patients with BOS Grade 3 manifested an acute exacerbation of airflow obstruction, which proved to be irreversible. CONCLUSIONS: Lung transplant patients with "flu-like" symptoms should proceed to IFA testing of NT swab specimens for early diagnosis. Samples collected within 7 days of symptom onset have high sensitivity as compared with serology and viral culture techniques. PMID- 12581765 TI - Early experience with sirolimus in lung transplant recipients with chronic allograft rejection. AB - BACKGROUND: Chronic lung allograft rejection, commonly manifest as obliterative bronchiolitis (OB/BOS), hinders long-term survival after lung transplantation (LT). OB/BOS is traditionally treated with augmented immunosuppression and results in short-term stabilization in pulmonary function for most patients. However, peribronchiolar fibroproliferation and airway obstruction usually recur despite initial improvements seen with increases in immunosuppression. In this observational, uncontrolled study, the effect of sirolimus, a novel immunosuppressant with anti-proliferative activity, was assessed in LT patients with OB/BOS. METHODS: Between June 1999 to November 2000, LT recipients with newly diagnosed or progressive OB/BOS received sirolimus in combination with a calcineurin inhibitor (CI) and prednisone. Pulmonary function, laboratory data and adverse effects were monitored for the first 24 weeks of therapy. RESULTS: Sirolimus was utilized in 12 LT recipients with OB/BOS. After drug initiation, 58% of patients required a reduction in CI dose to maintain appropriate CI trough concentrations. Despite CI dose reduction, serum creatinine rose in 75% of patients. Unexpected adverse effects included anemia of chronic disease (100%), edema (50%) and malignancy (17%). For the group, the rate of change in FEV(1) and FEF(25%-75%) was unchanged with sirolimus, but individual responses varied. CONCLUSIONS: For the group, the decline in pulmonary function was not affected by the addition of sirolimus. However, among individuals with rapidly declining pulmonary mechanics, sirolimus resulted in stabilization or improvement in pulmonary function. Significant adverse effects resulted from combination sirolimus plus CI therapy. Until optimal dosing strategies and a more complete adverse effect profile are established, combination therapy should be utilized cautiously in these patients. PMID- 12581767 TI - NOS substrate during cardioplegic arrest and cold storage decreases stunning after heart transplantation in a rat model. AB - BACKGROUND: In this study, we evaluated how adding L-arginine to Centre de Resonance Magnetique Biologique et Medicale (CRMBM) solution affected myocardial performance during post-ischemic in vivo reperfusion. METHODS: Experiments were conducted using a modified Lewis-Lewis heterotopic heart transplantation model, with a total ischemic time of 3 hours followed by 1 or 24 hours of blood reperfusion. Heart grafts were arrested using intra-aortic injection of CRMBM solution, either supplemented or not supplemented with 2 mmol/liter L-arginine (n = 12 in each group). We measured systolic indexes and simultaneously performed phosphorus magnetic resonance spectroscopy ((31)P MRS). We quantified total endothelial nitric oxide synthase (eNOS) protein using the Western blot test of freeze-clamped hearts. RESULTS: Contractility during early reperfusion was significantly better in grafts arrested with CRMBM solution enriched with L arginine: mean rate pressure product, 11249 +/- 1548 vs 5637 +/- 1118 mm Hg/min (p = 0.05), and maximal first derivative of the pressure signal (dP/dt(max)), 1721 +/- 177 vs 1214 +/- 321 mm Hg/sec (p = 0.013). Conversely, during late reperfusion, contractility did not relate to the nature of the preservation solution. The presence of L-arginine in the CRMBM solution did not alter time related variations of high-energy phosphate ratios measured using in vivo (31)P MRS. The eNOS protein level decreased significantly during early compared with late reperfusion, with no effect caused by L-arginine. CONCLUSIONS: During early reperfusion, the limited myocardial stunning observed with CRMBM solution containing L-arginine does not relate to energy metabolism but to better preservation of the NO pathway. PMID- 12581766 TI - CD8(+) T cells induce graft vascular occlusion in a CD40 knockout donor/recipient combination. AB - BACKGROUND: It has recently been shown that treatment of animals with antibodies to CD154 (CD40L), allows for prolongation of cardiac allograft survival, but does not inhibit development of graft vasculopathy. CD8(+) T cells have been implicated in this effect. In this study we assess the role of CD40-CD154 interactions and CD40-independent CD8(+) T cells in the permanent and complete absence of CD40 by using donors and recipients genetically deficient in CD40. METHODS: Hearts from BALB/c CD40(-/-) donors were transplanted into C57BL/6 CD40( /-) recipients in the presence or absence of CD8(+) T-cell depletion. At Day 60, hearts were examined for vasculopathy using quantitative morphometry and numbers of infiltrating T cells were counted. The intragraft expression of interferon gamma (IFN-gamma), transforming growth factor-beta1 (TGF-beta1), interleukin-4 (IL-4), eotaxin and CCR3 was assessed using competitive reverse transcription polymerase chain reaction (RT-PCR). RESULTS: In the absence of CD8(+) T-cell depletion, the mean percent intimal occlusion was 28% (with 50% of vessels showing no intimal occlusion). This figure was reduced significantly to 12% and 80% of vessels showing no intimal occlusion in mice receiving anti-CD8 antibody. Depletion of CD8(+) T cells was associated with significantly reduced intragraft IFN-gamma, TGF-beta1 and CCR3 expression, whereas mRNA production of IL-4 and eotaxin was increased. CONCLUSION: Vascular intimal occlusion progresses in the complete absence of CD40-CD154 interactions, albeit to quite a small degree. The residual disease is significantly reduced by anti CD8(+) T-cell treatment, confirming the importance of CD40-CD154-independent CD8(+) T cells in the genesis of this disease. PMID- 12581768 TI - Creatine kinase and troponin after myocardial preservation using HTK solution (Custoidol) for clinical heart transplantation. AB - Myocardial preservation by Bretschneider histidine-tryptophan-ketoglutarate (HTK) solution (Custoidol) was studied in transplanted patients. Post-operative creatine kinase (CK), CK-MB and troponin I were higher in patients with an ischemic time of >or=4 hours; however, values were within the acceptable range for clinical heart transplantation. PMID- 12581769 TI - Risk factors and management of bleeding associated with transbronchial lung biopsy in lung transplant recipients. AB - The aim of this study was to assess specific risk factors associated with bleeding during transbronchial biopsy (TBBx) in lung transplant recipients. Risk factors analyzed included gender, type of transplant, acute rejection, bronchiolitis obliterans syndrome (BOS) status, infections, number of biopsies obtained per procedures, serum creatinine level and post-operative day since transplantation. The bronchoscope was not wedged to obtain TBBx and associated bleeding was managed using the "back-and-forth" technique. The severity of bleeding is shown to be independent of any specific risk factor and the back-and forth technique described herein can be safely employed in lung transplant recipients to manage bleeding associated with TBBx performed without wedging of the bronchoscope. PMID- 12581770 TI - Improved systemic ventricular function after carvedilol administration in a patient with congenitally corrected transposition of the great arteries. AB - Congenitally corrected transposition of the great arteries (CCTGA) is associated with shortened survival due, at least in part, to progressive systolic dysfunction of the systemic ventricle. We report a substantial improvement in systemic ventricular function with carvedilol in a 63-year-old man with CCTGA. PMID- 12581771 TI - Assessment of myocardial recovery in a patient with acute myocarditis supported with a left ventricular assist device: a case report. AB - Acute myocarditis may present with profound hemodynamic compromise; however, spontaneous resolution of the inflammatory process may occur in up to half of such patients. In patients with fulminant myocarditis, mechanical circulatory support may serve as a bridge to myocardial recovery. In this report we describe a 35-year-old man with acute myocarditis who required left ventricular assist device support as a bridge to recovery, and suggest a method for determining the suitability and timing of device explantation. A combination of echocardiography, right heart catheterization, exercise testing and serial endomyocardial biopsies was used to determine the resolution of myocarditis, recovery of myocardial function and timing for device explantation. Successful device explantation was performed after 37 days of device support. Further study is required to assess the role of ventricular assist devices in combination with immunosuppressive therapy in the management of fulminant myocarditis. PMID- 12581772 TI - Interstitial pneumonitis associated with sirolimus: a dilemma for lung transplantation. AB - Rapamycin/sirolimus (SR), trade named Rapammune (Wyeth-Ayerst, Sydney, Australia), is a potent immunosuppressive drug associated with myelosuppression, hypertension, hyperlipidemia, and infection. Rapamycin/sirolimus-induced pneumonitis has been described previously in renal transplant recipients, and this report describes a stable heart-lung transplant recipient who developed a pulmonary infiltrate that reversed after ceasing SR therapy. We believe that immunosuppression-induced pneumonitis in a lung allograft is a serious dilemma for lung transplant physicians PMID- 12581773 TI - Successful treatment of enterovirus-induced myocarditis with interferon-alpha. AB - No randomized, placebo-controlled studies have investigated interferon-alpha therapy in enterovirus-proven myocarditis. This report describes 2 patients with enterovirus-induced myocarditis (1 with associated Churg-Strauss syndrome) who at follow-up endomyocardial biopsy showed clinical and hemodynamic improvement and viral clearance (using polymerase chain reaction) after interferon-alpha therapy. PMID- 12581774 TI - Stability indicating methods for the determination of aceclofenac. AB - Five new selective, precise and accurate methods for the determination of aceclofenac in the presence of its degradation product; diclofenac are described. Method A utilizes third derivative spectrophotometry at 242 nm. Method B is RSD(1) spectrophotometric method based on the simultaneous use of the first derivative of ratio spectra and measurement at 245 nm. Method C is a pH-induced difference (deltaA) spectrophotometry using UV measurement at 273 nm. Method D is a spectrodensitometric one, which depends on the quantitative densitometric evaluation of thin layer chromatogram of aceclofenac at 275 nm. Method E is RP HPLC that depends on using methanol: water (60:40 v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 275 nm. Regression analysis of a beer's plot showed good correlation in the concentration ranges 5-40, 10-40, 15-50, 50-200, 1 50 microg/ml for methods A, B, C, D and E, respectively. These methods are suitable as stability indicating methods for the determination of aceclofenac in presence of its main degradation product, diclofenac. The proposed methods were applied for the analysis of the drug in its pharmaceutical formulation and the results obtained were compared with those obtained with the official B.P. method. PMID- 12581775 TI - Preparation and in vitro evaluation of modified release ketoprofen microsponges. AB - Microsponges containing ketoprofen and Eudragit RS 100 were prepared by quasi emulsion solvent diffusion method. The effects of different mixing speeds, drug polymer ratios, solvent-polymer ratios on the physical characteristics of the microsponges as well as the in vitro release rate of the drug from the microsponges were investigated. All the factors studied had an influence on the physical characteristics of the microsponges. In vitro dissolution results showed that the release rate of ketoprofen was modified in all formulations. PMID- 12581776 TI - Binding modes of noncompetitive AMPA antagonists: a computational approach. AB - The activity of functional AMPA receptors (AMPARs) is modulated by noncompetitive antagonists. So far, no information about the molecular mechanism of action and the localization of the binding pocket(s) is available. We speculated that the leucine/isoleucine/valine binding protein (LIVBP)-like domain of AMPAR, localized at the extracellular N-terminus of the receptor, might be involved in the binding of noncompetitive antagonists and we tested this hypothesis through a computational approach involving the comparison with NMDA and metabotropic glutamate receptors and the generation of a 3D homology model of the LIVBP-like domain of AMPAR. The results suggest that the interdomain cleft of the LIVBP-like domain of AMPAR may contain the noncompetitive antagonist binding pocket. PMID- 12581777 TI - Synthesis and anti-HIV activity of 2,3-diaryl-1,3-thiazolidin-4-ones. AB - Several 1,3-thiazolidin-4-ones bearing a 2,6-dihalophenyl group at C-2 and a variously substituted phenyl ring at N-3 have been synthesized and tested as anti HIV agents. The results of the in vitro tests showed that some of them proved to be effective inhibitors of HIV-1 replication. PMID- 12581778 TI - Synthesis of 3,3'-(1-piperidino)substituted methylene-bis-isoxazoles preventing stimulus-induced leukocytes activation. AB - Some 3,3'-(1-piperidino)substituted methylene-bis-isoxazoles were prepared via Mannich base and tested to verify their antiinflammatory-related activity. Human neutrophils stimulated with either PMA and f-MLP were used as the cellular model. The efficiency of eight differently substituted compounds (2-9) was established on their capacity to reduce the O(2)(-) production by activated human neutrophils. The rising hydrophobicity in the side-chain of methylene-bis isoxazoles leads to a distinction in the neutrophil response against the two stimuli, favoring the inhibition of the PMA elicited cell activation and leaving inaffected the f-MLP induced cell responses. Compounds 8 and 9 are particularly active and abolish almost completely the neutrophil activation in the presence of PMA stimulus. PMID- 12581779 TI - Synthesis and biological evaluation of pyridazino[4,3-b]indoles and indeno[1,2 c]pyridazines as new ligands of central and peripheral benzodiazepine receptors. AB - A large number of pyridazino[4,3-b]indoles and indeno[1,2-c]pyridazines were synthesised and tested to evaluate their binding affinities at both central (CBR) and peripheral (PBR) benzodiazepine receptors. Relatively good PBR binding affinities were found for ligands belonging to the 3-arylmethyloxy pyridazinoindole series, whereas only 2-aryl-indenopyridazines 7a, 8a and 10a display a weak binding affinity for CBR. To find out the main structural determinants affecting PBR affinity, a molecular modelling study based on the comparative analysis of the three-dimensional properties of four properly selected derivatives 24a, 3b, 18a and 10d, with those of highly active and selective PBR ligands, taken as reference, was performed. PMID- 12581780 TI - Sensitive spectrophotometric determination of amlodipine and felodipine using iron(III) and ferricyanide. AB - A simple, accurate, sensitive and economical procedure for the estimation of amlodipine besylate and felodipine, both in pure form and in formulations has been developed. The method is based on the reduction of iron(III) by the studied drugs in acid medium and subsequent interaction of iron(II) with ferricyanide to form Prussian blue. The product exhibits absorption maximum at 760 nm in both cases. Beer's law is obeyed in the concentration ranges 5.0-15.0 and 1.5-5.0 microg/ml, for amlodipine and felodipine, respectively. The molar absorptivities are 1.76 x 10(4) and 4.24 x 10(4) l/mol cm. The corresponding Sandell sensitivities are 23.18 and 9.06 ng/cm(2). The limits of detection as well as quantification are reported. Seven replicate analyses of solutions containing three different concentrations of each drug were carried out and the percent error and the RSD values have been reported. The proposed method was applied to the determination of these drugs in pharmaceutical formulations and the results demonstrate that the method is equally accurate and precise as the official methods as found from the t- and F-values. The reliability of the method was established by recovery studies using standard-addition technique. PMID- 12581781 TI - Synthesis and anti-HIV activity of new C2 symmetric derivatives designed as HIV-1 protease inhibitors. AB - The synthesis of several new anti-HIV-1 compounds is described. The new compounds contain a C(2) symmetry axis and a dihidroxyethylene moiety based on the D tartaric acid back bone. The synthesis of these compounds was achieved in 36-69% overall yields from D-tartaric acid. The protocol included: acetylation of hydroxyl groups, followed by diamide formation and deacetylation or reduction with LiAlH(4). The anti-HIV 1 activities of these substances were evaluated in PM 1 cells, using Indinavir as standard (IC(50) = 0.2 microM). Two amino alcohol derivatives showed good inhibitory activity against the virus, with IC(50) = 2.0 and 4 microM. PMID- 12581782 TI - Polarographic behavior and determination of finasteride. AB - The polarographic behavior of finasteride at the dropping mercury electrode (DME) was studied adopting direct current (DC(t)), alternating current (AC(t)) and differential-pulse polarography (DPP) modes. In Britton-Robinson buffer (BRb), finasteride exhibited cathodic waves over the pH range 6-12. At pH 10, a well defined cathodic wave was obtained. The latter could be characterized as being irreversible, diffusion-controlled and partially affected by adsorption phenomenon. The number of electrons involved in the reduction process was accomplished and a proposal of the electrode reaction was presented. The current concentration plots were rectilinear over the ranges 8-40 and 2-30 microg ml(-1) using DC(t) and DPP modes, respectively. The minimum delectability was 0.2 microg ml(-1) (5.4 x 10(-7) M), for the latter. The proposed method was successfully applied to the determination of finasteride in its commercial capsules and the results obtained were in good agreement with those given with a reference method. PMID- 12581784 TI - Making gerontology popular. PMID- 12581783 TI - A new spectrofluorimetric method for the determination of lisinopril in tablets. AB - An accurate and precise spectrofluorimetric method is presented for the determination of lisinopril based on the formation of a derivative formed with 7 chloro-4-nitrobenzofurazan. The derivatization reaction proceeds quantitatively at pH 8.5-9.0 and 60 degrees C in 70 min when the molar ratio of reagent to the drug is 170. After the extraction with ethyl acetate the fluorescence intensity of the derivative was measured at 528 nm with excitation at 465 nm. Calibration graph is rectilinear over the range of 50-1000 ng/ml with detection and determination limits of 20 and 50 ng/ml, respectively. The regression equation is I(f) = 0.198C-0.299 (r = 0.9999). The method was applied to the commercially available tablets and the results were statistically compared with those obtained by official HPLC method. PMID- 12581785 TI - Does overexpression of Cu,Zn-SOD extend life span in Drosophila melanogaster? AB - The argument is presented here that, while there exists considerable correlative evidence supporting the oxidative stress hypothesis of aging, key experiments to test this hypothesis using Drosophila transgenics overexpressing Cu,Zn-SOD fall short in providing validation. In particular, significant life span extensions tend to be observed when the backgrounds of the control flies are relatively short-lived, suggesting that bolstering antioxidative defenses is only effective in compromised genetic backgrounds. Other shortcomings of the transgenic approaches adopted to date will be summarized. PMID- 12581786 TI - Signal transduction in T lymphocytes and aging. AB - Subclasses of cells in different compartments of the immune system possesses all those attributes, that make them suitable though somewhat limited models for the investigation of cellular processes during aging. Blood samples provide relative easily high amount of cells belonging to the same subclass, all of them having complex cascade processes in their signal transduction mechanisms, therefore being excellent targets for such investigations. One such subclass comprises peripheral blood lymphocytes. The signal-transduction cascade across the plasma membrane of lymphocytes displays many of the general features enabling us to draw conclusions for other cellular signaling problems that may arise during aging in other cell types not directly related to the immune system. The advantage of this approach lies in the fact that sometimes it is extremely difficult to study signal transduction processes in certain cell types under physiological conditions. The simultaneous occurrence of physical, chemical and molecular biological regulation of the immune processes at cellular and network levels make them very good examples for focusing our interest also on similar processes in other systems and cells. The fast developing new measuring techniques and the rapidly accumulating experimental data make it relatively easy to provide interesting new aspects, and ideas in this field. Finally, the immune system itself has its great importance and after all, it has an obvious declination with aging, the immune-senescence. PMID- 12581787 TI - Effects of renin-angiotensin system blockade in the aging kidney. PMID- 12581788 TI - A novel putative collaborator of p19ARF. AB - We performed a yeast interactive screen in search of p19(ARF)-binding partners and have isolated a novel serine rich protein that is assigned to human chromosome 4q35 and mouse chromosome 8. The human and mouse proteins showed 84.2% homology. It is named CARF for its role as a putative Collaborator of ARF. CARF binds to both mouse and human ARF proteins. Its expression was detected in a variety of human tissues. The cDNA was expressed in bacteria and mammalian cells as a soluble and predominantly nucleoplasmic protein, respectively. CARF is a novel binding partner of ARF and might be involved in its p53-dependent or independent tumor suppressive functions. PMID- 12581789 TI - Influence of age and caloric restriction on liver glycolytic enzyme activities and metabolite concentrations in mice. AB - The influence of caloric restriction (CR) from 2 months of age on the activities of liver glycolytic enzymes and metabolite levels was studied in young and old mice. Livers were sampled 48 h after the last scheduled feeding time. Old mice on CR showed significant decreases in the activities of all the enzymes studied, except for aldolase, triosephosphate isomerase and phosphoglycerate mutase, which were unchanged. The metabolites glucose, glucose-6-phosphate, fructose-6 phosphate, pyruvate and lactate were lower while fructose-1,6-bisphosphate, glyceraldehyde-3-phosphate, dihydroxyacetone phosphate, 3-phosphoglycerate and phosphoenolpyruvate were increased in old CR. Young mice on CR also showed reduced enzyme activities, except for aldolase, triosephosphate isomerase and enolase which were unchanged when compared with young controls. The metabolites glucose, glucose-6-phosphate, fructose-6-phosphate and pyruvate were decreased when compared with young controls, while phosphoenolpyruvate was increased. Ketone bodies increased (65%) in old, but not young, CR mice while fructose-2,6 bisphosphate decreased in both young (22%) and old CR (28%) mice. The results indicate that decreased hepatic glucose levels in CR mice are associated with decreased enzyme activities but not a uniform decrease in metabolite levels. Increased ketone body levels indicate increased utilization of non-carbohydrate fuels while decreased fructose-2,6-bisphosphate level suggests its importance in the control of glycolysis in CR. PMID- 12581790 TI - Caloric restriction increases gluconeogenic and transaminase enzyme activities in mouse liver. AB - Long-term caloric restriction (CR) has been shown to extend maximum life span in laboratory rodents. We investigated the activities of gluconeogenic and transaminase enzymes in the livers of old and young mice fed either control or calorie-restricted diets. Livers were sampled 48 h after the last scheduled feeding time. Old mice on CR showed significant increases in the activities of pyruvate carboxylase, phosphoenolpyruvate carboxykinase, fructose-1,6 bisphosphatase and glucose-6-phosphatase when compared with controls, indicating increased gluconeogenesis. Increased activities of tyrosine, tryptophan, histidine, phenylalanine, alanine and aspartate transaminases, as well as of malate and glutamate dehydrogenases were also observed, while branched-chain amino acid transaminase was unchanged. Young mice on CR showed a significant increase only in the phosphoenolpyruvate carboxykinase activity in the gluconeogenic pathway, while transaminases were increased significantly, except for tryptophan and branched-chain amino acid transaminases. Glutamate dehydrogenase also showed increased activity but malate dehydrogenase was unchanged. Increases in the level of acetyl-CoA and [Acetyl-CoA]/[CoA] ratio were observed only in the old CR mice. Our results demonstrate increased gluconeogenic activity in CR mice and are consistent with a state of increased hepatic gluconeogenesis and protein turnover during CR. PMID- 12581791 TI - Accelerated aging of senescence accelerated mice R-1 demonstrated by flash visually evoked cortical potentials. AB - To determine the physiological alterations of visual functions induced by aging, the latency of the N40 peak of the flash visual evoked cortical potentials at several stimulus frequencies were analyzed from senescence accelerated mice (SAM). The senescence prone (P-8) and senescence resistant (R-1) SAM lines were studied. In both the P-8 and ICR (the standard outbred albino laboratory mouse also called CD-1) mice, the peak latency was not significantly different at 6 and 12 months of age. In contrast, there was a prolongation of the peak latency in the R-1 line at 12 months compared to that at 6 months. We conclude that there is an acceleration of the aging process in the R-1 line for visually evoked responses. Thus, the R-1 line might be an independent line suited for the study of aging effects on visual functions. PMID- 12581792 TI - Dietary vitamin E and physical exercise: I. Altered endurance capacity and plasma lipid profile in ageing rats. AB - The effect of vitamin E on the exercise performance and plasma lipid profile was studied in male Wistar rats of 4-(young adults), 8-(old adults), 12-(middle-age) and 22-months (old) of age. Animals were orally supplemented with vitamin E and allowed to swim for 30 min/day, 5 days/week and for a total period of 60 days. Swim velocity (S(v)), external work done (W(ext)) and endurance (E) capacity were the parameters that were used to assess the exercise performance of the trained rats that were either supplemented or non-supplemented with the dietary antioxidant. Plasma lipid profile analyses were in terms of low-density lipoprotein (LDL-C), high-density lipoprotein, (HDL-C) cholesterol and total cholesterol (C). Age-related decline in S(v) was noticeable in the 22-months old rats. However, the effect of vitamin E on the S(v) between the trained groups was not evident in any of the age groups. W(ext) increased linearly with age with no significant variations between the trainees. Trainee rats, when allowed to swim to exhaustion, showed a higher endurance capacity when supplemented with vitamin E. However, this capacity declined with age. There was a significant age associated elevation in plasma C with corresponding increase in LDL-C. Exercise training in conjunction with vitamin E supplementation was most effective in elevating HDL-C levels in all age groups. These changes were accompanied by significant reductions in cholesterol/HDL-C ratios in animals receiving vitamin E, sedentary or otherwise. Our data suggests that it may be important to consider vitamin E while attempting to derive the benefits of swim training, both in terms of favorably altering the plasma lipid profile as well as enhancing the endurance capacity of exercise trainees. Dietary supplementation by vitamin E could attenuate the early onset of fatigue in the old. PMID- 12581793 TI - Dietary vitamin E and physical exercise: II. Antioxidant status and lipofuscin like substances in aging rat heart. AB - The heart faces a high risk of free radical injury owing to a slow generation of antioxidant (AO) enzymes by its cells. A general decline in this system may be another reason for the development of age-related diseases. Although the correlation between aging and exercise has been studied extensively, these studies have produced conflicting data on the effects of vitamin E on the aging heart, when it is introduced as an intervening factor. To investigate these effects, we determined the activities of antioxidant enzymes (AOEs) such as superoxide dismutase (SOD) and catalase (CAT), lipid peroxidation (LP), lipofuscin (LF)-like autofluorescent substances and vitamin E content in the left and right ventricles (LV and RV) of the heart in male Wistar albino rats of 4 (young adults), 8-(old adults), 12-(middle-age) and 22-mos(old) of age. Animals were orally supplemented with vitamin E and allowed to swim for 30 min/day, 5 days/week and for a total period of 60 days. Exercise training in all the age groups except the old was effective in upregulating the SOD activity. Old trainees showed an increase in SOD activity when supplemented with vitamin E. In the 22-mo-olds, a remarkable decrease in CAT activity was seen. Exercise by itself upregulated the CAT as well as SOD activity in all age groups except the old wherein vitamin E was effective in increasing the activities of AOEs. Supplementation significantly reduced LP as evidenced by lowered malondialdehyde (MDA) and LF-like autofluorescent substances in the trained as well as sedentary rats. Tissue vitamin E content was low in the swim trainees that were not supplemented. This change, well emphasized in the trainee groups of 22-mo-old suggests the probable utilization of vitamin E in keeping free radicals at bay. Our results suggest that vitamin E can stand out as a significant tool in ameliorating the declining AO defense in the old rats. PMID- 12581794 TI - Preconditioning with heat shock further improved functional recovery in young adult but not in middle-aged rat hearts. AB - Ischemic preconditioning (PC) improves post-ischemic function, and heat shock (HS) mimics delayed PC in young animals. However, PC is not protective and the consequences of HS are not known in the aging hearts. This report examines the efficacy of HS and its synergy with PC in the middle-aged rat hearts. Hearts from 12- or 50-week-old rats were subjected to PC before 25 min ischemia followed by 30 min reperfusion 48 h after HS. HS induced HS proteins (HSP) in both age groups but that PC and HS translocated PKC-alpha and -delta only in young rats. The beneficial effects of HS and PC were additive and enhanced protein kinase C (PKC) translocation in young rats. However, neither HS alone nor in combination with PC conferred any functional advantage or accelerated PKC translocation in old rats. Similarly neither HS alone nor in combination with PC restore PC effects in old rats with impaired PKC activation, despite the induction of HSP, indicating that induction of HSP is insufficient for cytoprotection. PMID- 12581795 TI - Developmental and age-related alterations of calcium homeostasis in human fibroblasts. AB - Calcium homeostasis, in terms of both cytosolic calcium concentration ([Ca(2+)](i)) and capacitative calcium entry, has been investigated in human skin and lung fibroblasts at different developmental and ageing stages by employing the Fura-2 based measurements. [Ca(2+)](i) in foetal lung or skin cells were nearly similar. However, significant changes were observed between foetal and adult fibroblasts and interestingly in opposite directions depending on the tissue of origin. In particular, in adult lung cells [Ca(2+)](i) was more than three-fold higher compared to adult skin fibroblasts, a difference which may contribute to tissue-specific functions. Capacitative Ca(2+) entry, i.e. the transient [Ca(2+)](i) increase induced by re-addition of extracellular calcium after depletion of thapsigargin-sensitive calcium stores, was found to exhibit the same pattern of differences during foetal-to-adult transition (it is two-fold higher in adult lung cells than in adult skin cells). At variance, we found capacitative Ca(2+) entry to be significantly attenuated during in vivo or in vitro ageing of fibroblasts; while minor alterations of [Ca(2+)](i) were observed. These findings indicate that capacitative calcium entry rather than [Ca(2+)](i), is mainly affected during the ageing process. PMID- 12581797 TI - Correlation between accelerated presbycusis and decreased immune functions. AB - The aim of the current study is to analyze the relationship between presbycusis and the immune system, which is affected by pathogenic environments, and to devise a strategy for the prevention of presbycusis using the SAMP1 mouse, an animal model for accelerated senescence that shows both immunological dysfunction and hearing loss caused by the impairment of spiral ganglion cells in the cochlea. When these mice were bred in different pathogenic environments, we found that the development of age-related diseases such as presbycusis was delayed in the mice bred under clean conditions. Prednisolone administration showed no significant prevention of the development of presbycusis in the mice, suggesting that autoimmune mechanisms are not involved in the acceleration of presbycusis. It is conceivable that pathogen-induced infections impose a severe stress on the host, impairing the host's immune functions. A reduction in the number of pathogens may therefore prevent the acceleration of the aging process. These findings suggest that not only the gene backgrounds but also immune functions affect the development of presbycusis in SAMP1 mice. Further studies into the relationship between systemic immune functions and the neuro-generation system may provide additional information about the treatment for age-related diseases. PMID- 12581796 TI - Association between the HLA-DR alleles and longevity: a study in Sardinian population. AB - Human longevity may be correlated with optimal functioning of the immune system, suggesting that genetic determinants of longevity also resides in those polymorphisms for the immune system genes that regulate immune responses as histocompatibility (HLA) antigens. However, conflicting results have been obtained. Some well planned and designed association studies performed in Caucasians suggest that longevity is associated with positive selection of alleles (i.e. HLA-DR11) or haplotypes (i.e. HLA-B8,DR3) that confer resistance to infectious diseases, respectively, via peptide presentation or via antigen non specific control of immune response. Association studies are subjected to a number of possible confounding factors, the homogeneity of the population in term of geographical origin among others. Because of the lack of large-scale heterogeneity, the Sardinians represent a suitable population for association studies addressed to dissect the complex traits as longevity. Thus, we have evaluated, by the amplification refractory mutation system/polymerase chain reaction, HLA-DR frequencies in 120 centenarians (79 women and 41 men) and 86 controls (53 women and 33 men) from Sardinia, to validate, in this very homogeneous population, the associations between HLA alleles or haplotypes and longevity observed in other Caucasoid populations. No significant differences were obtained by analysing the differences between Centenarians and controls except for HLA-DRB1*15 that was increased in centenarians. However, the significance was not maintained by multiplying P values for the number of alleles under study. Thus, in Sardinian centenarians, we were not able to confirm the findings observed in the well planned and designed studies performed in other Caucasoid populations. Besides, HLA HFE gene polymorphisms have been recently demonstrated to be associated with longevity in the Sicilian population but not in Danish one. On the whole these findings clearly show that HLA/longevity associations are population-specific, being heavily affected by the population specific genetic and environmental history. So, in our opinion, HLA genes might be considered survival genes not longevity genes. PMID- 12581798 TI - Regional variability in mottled subclinical melanoderma in the elderly. AB - The density in melanin chromatophores becomes heterogeneous in the epidermis during the early events of photoaging. Little is known about the regional variability in the resulting mottled skin appearance on the sun-exposed parts of the body in the elderly. The relationship between these features and the dermal atrophy related to aging is also an area ripe for study. The aim of the present study was to objectively assess and compare such aspects of photoaging in older subjects. A computer-assisted video camera equipped with an internal ultraviolet emitting unit (Visioscan) was used in combination with image analysis to quantify the infraclinical mottling of the skin on the forehead, neck and dorsal forearm. A Densi Score device served to assess the dermal atrophy by rating the skin folding capacity. A mottled subclinical melanoderma was disclosed on the three evaluation sites in all subjects irrespective of their phototypes. Three main patterns were identified corresponding to regular perifollicular dots, streaky macules along shallow wrinkles and unevenly shaped macules in the interfollicular area. The extent in melanoderma was larger on the forehead than on the neck and forearm. The age after 60 years did not influence the severity of mottled melanoderma, but clearly altered the skin folding capacity. The greatest inter individual differences in mottled melanoderma were found in phototype II subjects. In conclusion, infraclinical mottled melanoderma is present in all individuals after 60 years of age. This type of epidermal photoaging does not appear to be influenced in older ages. By contrast, skin laxity due to dermal aging continues to progress beyond the age of 60 years. PMID- 12581799 TI - Genome size and longevity in fish. AB - The wide variety of genome sizes (measured as C-value) observed across taxa is not related to organismal complexity or number of coding genes. Partial answers to this C-value enigma have been found by establishing associations between C value and particular phenotypic characteristics. One such controversial association has been recently suggested between genome size and longevity in birds. In order to determine whether genome size is a general predictor of longevity, we have extended the analysis to the Actinoptergyian fish, a widely divergent group in terms of both longevity and genome size. We collected data on genome size, longevity and body mass for species covering fourteen orders of bony fish. Analysis of covariance using order as a cofactor shows a significant effect of genome size on longevity (corrected for body mass), with lifespan increasing as a function of genome size. Analysis of phylogenetically independent contrasts for orders with a large number of species with a well resolved phylogenetic relationship (Acipenseriformes, Cypriniformes, and Salmoniformes) found the same trend of longer lifespan with increases in genome size but the relationship was not significant. Our results consistently show an increase in lifespan for fish with larger genomes. PMID- 12581800 TI - The mitochondrial DNA A3243G mutation in Werner's syndrome. AB - OBJECTIVE: The contribution of the A3243G mutation in mitochondria DNA (mtDNA) to diabetes mellitus (DM) in Werner's syndrome (WS) was studied. PATIENTS AND METHOD: DNA samples from peripheral white blood cells (WBCs) originating from 24 Japanese WS patients aged 30-56 were used. For control, 239 subjects aged 15-95 were also used. The mtDNA was amplified using specific primers. After HaeIII digestion, the ratio of the A3243G mutation was compared. RESULTS: The ratio of the A3243G mutation is 0.45+/-0.13% in WS, which is statistically insignificant from those in the control groups at various age. The mutation types of WRN in genomic DNA did not affect the ratio of the A3243G mtDNA mutation. No significant difference was observed concerning to the ratios among the WS patients with and without DM, and also controls. Furthermore, no significant difference was observed in the ratios of A3243G mutation among controls from various age groups. CONCLUSION: The A3243G mutation in mtDNA does not accumulated in WBCs from WS. Mitochondria A3243G mutation may not contribute to the pathogenesis of DM observed in WS. PMID- 12581801 TI - The metabolic rate of Caenorhabditis elegans dauer larvae: comments on a recent paper by Houthoofd et al. PMID- 12581802 TI - C. elegans, a model for aging with high-throughput capacity. AB - The 1 mm long nematode Caenorhabditis elegans is one of the prime animal models to study the genetics of aging. Wild type animals under laboratory conditions live only for an average of 18 days, whereas mutations in about 50 genes have been identified that extend longevity up to sixfold. High-throughput analyses have been devised that allow large-scale analysis to identify additional genes, as well as compounds that affect life-span. PMID- 12581803 TI - Issues in the diagnosis of attention-deficit/hyperactivity disorder in children. AB - This paper provides a brief overview of the nature of attention deficit/hyperactivity disorder (ADHD) in children and the current criteria used in its clinical diagnosis. While the disorder continues to be viewed as one of inattention and/or hyperactive-impulsive behavior, theories of ADHD are beginning to focus more on poor inhibition and deficient executive functioning (self regulation) as being central to the disorder. Problems have been identified by research pertaining to the clinical diagnostic criteria outlined in the DSM-IV that, at present, remain unresolved. Clinicians should be aware of these problems and the adjustments that need to be made to them when dealing with special populations that were not represented in the field trials used to develop these criteria. PMID- 12581804 TI - West & West syndrome--a historical sketch about the eponymous doctor, his work and his family. AB - The British surgeon William James West has not left a tremendous literary or scientific work as many of his contemporaries did. For this reason only a little has been known about him and the fate of his family for decades, even though the eponym was created in the 1960s. Only in 1990 was a first biography published and later on supplemented. If his son had not suffered from the syndrome, which later on was named after him, he would not have published the first description of the West syndrome in The Lancet in 1841. Possibly we would be talking about Newnham's syndrome, because 8 years later he published a detailed report on this subject. There is, however, a second aspect concerning the pioneering activity of West, i.e. his advocating of ovariotomy in its early days. To judge the importance of this feat correctly, it is necessary to keep in mind that his former pupil and practice-partner Gorham in 1874 obviously tried to exaggerate the role of West in ovariotomy. Nevertheless, it is worth keeping the memory of William James West and his son James Edwin alive, as happened at the International Symposium on West Syndrome and Other Infantile Epileptic Encephalopathies at Tokyo, 9-11 February in 2001, and by other occasions. PMID- 12581805 TI - Neuron-specific enolase, nucleotides, nucleosides, purine bases, oxypurines and uric acid concentrations in cerebrospinal fluid of children with meningitis. AB - To determine the effects of meningitis on cerebral energy metabolism, cerebrospinal fluid concentrations of adenosine monophosphate, inosine monophosphate, inosine, adenosine, guanosine, adenine, guanine, hypoxanthine, xanthine and urate were determined by high-performance liquid chromatography, and neuron-specific enolase by an enzyme immunoassay method, in 100 children with meningitis (45 bacterial, 46 viral and nine tuberculous), aged between 1 month and 13 years, and in 160 age-matched controls. Compared with controls, patients with bacterial meningitis showed high concentrations of hypoxanthine, xanthine and urate; patients with viral meningitis showed high concentrations of inosine, guanosine, xanthine, urate and neuron-specific enolase; and patients with tuberculous meningitis showed very high concentrations of inosine, xanthine and urate. Xanthine and urate concentrations were significantly higher in patients with tuberculous meningitis than in patients with viral or bacterial meningitis. These results suggest that in the acute stage of bacterial, viral and tuberculous meningitis, neuronal energy metabolism may be altered. The measurement of cerebrospinal xanthine and uric acid concentrations may be useful for the early diagnosis of a tuberculous origin. PMID- 12581806 TI - Unilateral cortical necrosis following status epilepticus with hypoglycemia. AB - Isolated status epilepticus or severe hypoglycemia rarely causes irreversible focal neurologic deficits in children. We describe three children who presented with status epilepticus and prolonged hypoglycemia resulting in hemiplegia due to unilateral hemispheric damage. The non-vascular cortical topography of the lesions is consistent with selective neuronal necrosis, confirmed by histopathology in one patient. This suggests increased neuronal vulnerability to necrosis secondary to energy failure resulting from combination of hypoglycemia and status epilepticus. PMID- 12581807 TI - Effects of valproate and carbamazepine on serum levels of homocysteine, vitamin B12, and folic acid. AB - Homocysteine (HMC) is a sulfur containing amino acid, which plays a role in methionine metabolism. Folic acid (FA) and vitamin B12 (B12) are essential for remethylization of HMC to methionine. HMC level increases in the deficiency of these vitamins. Hyperhomocysteinemia causes vascular endothelial damage, which causes atherosclerosis. The aim of this study is to investigate the effect of valproate (VA) and carbamazepine (CBZ) on the serum levels of HMC, B12, and FA.Thirty-six children receiving CBZ and 30 children receiving VA for epilepsy for the last 1-year period and 29 healthy children as control were the population of this study. After 6 h of fasting serum HMC, B12, and FA levels were measured and results were compared statistically. Mean values of HMC, FA, and B12 levels in control group were 9.2+/-2.7 micromol/l, 9.0+/-2.0 ng/ml, and 342+/-162 pg/ml, in VA group 14.0+/-6.8 micromol/l, 7.3+/-2.9 ng/ml, and 368+/-159 pg/ml, in CBZ group 16.0+/-13.1 micromol/l, 7.5+/-3.3 ng/ml, and 285+/-158 pg/ml, respectively. Serum HMC levels were higher in VA and CBZ groups than control group (P<0.01 and P<0.05, respectively). Serum FA levels were lower in VA and CBZ groups compared to control group (P<0.05). Serum levels of B12 were not different between VA and control groups (P>0.05). In CBZ group serum B12 levels were lower than control group (P<0.05).FA may be added to the treatment protocol (if the patients take only CBZ, then B12 should also be added) for patients taking these antiepileptic drugs to decrease the degenerative effect of VA and CBZ on vascular endothelium. PMID- 12581808 TI - Molecular genetic study in Japanese patients with Alexander disease: a novel mutation, R79L. AB - Since the first report by Brenner et al. of mutations in the glial fibrillary acidic protein (GFAP) gene in patients with Alexander disease, several molecular genetic studies have been performed in different ethnic groups. We previously reported a Japanese patient with a mutation, R239C, which is identical to one commonly found in American patients. Here we have analyzed four additional Japanese patients by screening for known mutations or, if no known mutation was found, by sequencing of all exons of the GFAP gene. We detected three missense mutations; one was a novel mutation, R79L, and two were previously reported mutations, R239C and R79C. All of our patients were heterozygous for their mutations. Together with the novel mutation, R79L, four different nucleotide changes altering the R79 residue have been reported, implying that any alternation of this arginine residue can give the GFAP protein a dominant negative effect, leading to accumulation of GFAP as Rosenthal fibers. We conclude that molecular genetic analysis of the GFAP gene is feasible for antemortem diagnosis of Alexander disease in Japanese patients. PMID- 12581810 TI - Carbamazepine-induced abnormal pitch perception. AB - A 7-year-old boy began to complain that his pitch perception was decreased just after oral medication with carbamazepine was initiated for the treatment of epilepsy. When he played the piano, he felt as if he had played a musical note of almost a half pitch lower than he had. His pitch perception recovered soon after the cessation of carbamazepine. A 14-year-old girl noted a lowered pitch of music sounds while she played the piano just after the administration of carbamazepine for the treatment of epilepsy. Carbamazepine was withdrawn and the auditory symptoms disappeared. Both patients were musically trained. Reversible pitch perception abnormalities are a rare adverse effect of carbamazepine, however, the clinical features of the reported cases were similar; they were musically trained, young, female and Japanese. Although the mechanism remains unclear, we have to pay attention to this subtle adverse effect when we treat epileptic patients with carbamazepine. PMID- 12581809 TI - Bilateral posterior agyria-pachygyria and epilepsy. AB - We analyzed the electroclinical findings in two patients with bilateral posterior agyria-pachygyria. Both patients presented with mental retardation, mild motor deficit and epilepsy. The electroclinical findings were characterized by frequent tonic or atonic generalized seizures with occasionally simple or complex partial seizures. Interictal electroencephalography (EEG) showed occipital spikes and diffuse polyspike-wave paroxysms predominantly in the posterior region. Ictal EEG showed diffuse 10-11 Hz activity. Cerebral magnetic resonance imagings (MRIs) showed thickened cortex in the parieto-occipital lobes, bilaterally and symmetrically. The volume of underlying white matter appeared reduced, and the overlying subarachnoid spaces were enlarged. The occipital horns were dilated. These findings were compatible with agyria-pachygyria of the posterior portions of the brain. In conclusion, in patients with mental retardation, mild motor deficit and epilepsy characterized by tonic or atonic generalized seizures, interictal EEG with diffuse polyspike-wave paroxysms predominantly in posterior region, posterior focal epileptilorm abnormalities and ictal diffuse 10-11 Hz activity, bilateral parieto-occipital agyria-pachygyria should be considered as a possible etiology. Magnetic resonance image is the best neuroradiological study to identify this disorder of cortical development. PMID- 12581811 TI - Microcephaly with plate-like cortical calcification. AB - Congenital malformations of the central nervous system have varied presentations depending on the stage of embryological development when the insult occurred. We report a neonate with microcephaly associated with plate-like cortical calcification, probably secondary to an unidentified insult at around 11-12 weeks post-conceptional age causing an arrest of the proliferative phase. The associated vasculopathy probably caused diffuse cortical calcification. PMID- 12581812 TI - Congenital asymmetric crying facies and agenesis of corpus callosum. AB - Although association of congenital asymmetric crying facies (CACF) with major congenital anomalies of central nervous system (CNS) has been described, brain magnetic resonance imaging (MRI) studies have not been reported. Two children who had CACF associated with agenesis of corpus callosum (ACC) diagnosed by MRI are described. Neurofibromatosis type 1 (NF-1) was diagnosed in one case. Both patients had developmental delay. To the best of our knowledge, only one previous case with CACF associated with ACC has been reported, but our cases are the first cases reported with the characteristic findings of ACC on MRI. Although cafe-au lait spots have been described in previous cases, the coexistence of CACF and NF 1 has not previously been reported. Although these associations may be coincidental, clinicians should be aware of the potential link between these entities. Furthermore, these findings emphasize the importance of MRI studies for detecting brain anomalies in cases with CACF and suspected CNS involvement. PMID- 12581813 TI - Severe generalized dystonia induced by metoclopramide in a girl with methylmalonic acidemia. PMID- 12581814 TI - Anal protrusion of ventriculo-peritoneal shunt catheter in an infant. PMID- 12581815 TI - Somatic panic-attack equivalents in a community sample of Rwandan widows who survived the 1994 genocide. AB - The present study is the first to attempt to determine rates of panic attacks, especially 'somatically focused' panic attacks, panic disorder, symptoms of post traumatic stress disorder (PTSD), and depression levels in a population of Rwandans traumatized by the 1994 genocide. The following measures were utilized: the Rwandan Panic-Disorder Survey (RPDS); the Beck Depression Inventory (BDI); the Harvard Trauma Questionnaire (HTQ); and the PTSD Checklist (PCL). Forty of 100 Rwandan widows suffered somatically focused panic attacks during the previous 4 weeks. Thirty-five (87%) of those having panic attacks suffered panic disorder, making the rate of panic disorder for the entire sample 35%. Rwandan widows with panic attacks had greater psychopathology on all measures. Somatically focused panic-attack subtypes seem to constitute a key response to trauma in the Rwandan population. Future studies of traumatized non-Western populations should carefully assess not only somatoform disorder but also somatically focused panic attacks. PMID- 12581816 TI - Obsessive-compulsive behaviors in parents of multiplex autism families. AB - Parents of autistic probands with high and low rates of repetitive behaviors were compared for rates of obsessive-compulsive traits and disorder. The rate of repetitive behaviors was assessed using the Autism Diagnostic Interview-Revised (ADI-R) in 176 autistic probands from 57 multiplex families. Obsessive-compulsive disorder (OCD) in parents was determined by direct interview using a parental history questionnaire, with screening for obsessive-compulsive traits using the Yale-Brown Obsessive-Compulsive Scale checklist. Children who had high total scores on the repetitive behavior domain of the ADI-R were significantly more likely to have one or both parents with obsessive-compulsive traits or disorder compared with children who had low total scores on this domain. Children with high scores on D1/D2 of the ADI-R (narrow restricted interests and rituals) were significantly more likely to have one or both parents with OCD, especially fathers, than those with low D1/D2. The occurrence of obsessive-compulsive traits or disorder in parents of autistic children in multiplex families is significantly more likely if autistic children have a high occurrence of repetitive behaviors. Dichotomizing autistic probands by severity and type of repetitive behaviors (circumscribed interests and compulsive rituals) may yield more homogenous groups, which could be helpful in genetic linkage studies. PMID- 12581817 TI - Qualitative analysis of verbal fluency in depression. AB - The aim of this study was to analyze qualitative aspects of verbal fluency in depression. Phonemic and semantic output was scored for word clustering and switching between clusters in depressed patients and normal control subjects. Depressed patients (n=25) and normal control subjects (n=19) were administered both phonemic and semantic fluency tasks. All patients were also evaluated with executive card sorting tests. Patients with depression produced fewer words on the semantic fluency task than controls and showed normal performance on the phonemic fluency tasks. The deficit on semantic fluency of depressed patients was related to a reduced number of switches with normal cluster sizes. The number of switches in depression was associated with a reduced ability to shift mental set on card sorting tests, suggesting that verbal fluency impairment reflects general executive problems in depression. PMID- 12581818 TI - Verbal learning and memory in schizophrenic and Parkinson's disease patients. AB - The purpose of this study was to investigate the neurofunctional substrate of verbal learning and memory impairments in schizophrenic patients. In this pilot study, our aim was to compare the memory disturbance of schizophrenic patients to the subcortico-frontal memory profile of Parkinson's disease (PD) patients. The California Verbal Learning Test, a verbal episodic memory test, was administered to 60 subjects, 20 patients with schizophrenia, 20 patients with PD and 20 healthy control subjects. All subjects were aged between 50 and 70 years and all patients were in a stable phase. Like the Parkinson patients, the schizophrenic patients showed a major deficit of retrieval characterized by deficit of recalls but contrarily to PD patients, schizophrenic patients' encoding scores were altered. These impairments in episodic memory could suggest a dysfunction of the subcortico-frontal circuits in schizophrenic patients. However, they demonstrated an additional encoding deficit associated with probable frontal in situ alteration. PMID- 12581819 TI - Reduced levels of specific autobiographical memories in schizophrenia. AB - Autobiographical memory is intrinsically related to the self and personal identity. This study investigated whether both personal episodic memory and semantic memory are impaired in schizophrenia, a disease characterized by an abnormal personal identity. Personal episodic memory and personal semantic memory were investigated in 24 patients with schizophrenia and 24 normal subjects using an autobiographical fluency task and an autobiographical memory inquiry. Autobiographical memory scores and the proportion of specific memories were lower in patients with schizophrenia than in normal subjects. The deficit of personal episodic and semantic memory, as assessed by the autobiographical memory inquiry and the autobiographical fluency task, respectively, was most apparent after the onset of clinical symptoms. Schizophrenia is associated with an impairment of both personal episodic and semantic memory and with a reduction of specific autobiographical memories. Those impairments are consistent with the existence of an abnormal personal identity in patients with schizophrenia. PMID- 12581820 TI - Depressive episodes in stable schizophrenia: critical evaluation of the DSM-IV and ICD-10 diagnostic criteria. AB - Depressive episodes are a common and potentially severe occurrence in schizophrenia but are poorly recognised by psychiatrists. Coherent diagnostic criteria are necessary to improve diagnosis and treatment of these conditions. To evaluate the usefulness of the ICD-10 category of post-schizophrenic depression (PSD) and the DSM-IV category of postpsychotic depressive disorder of schizophrenia (PDDS), 80 clinically stable schizophrenic outpatients were evaluated with two independent measures of depression, a dimensional measure and a categorical measure. One rater applied the DSM-IV criteria for major depressive episodes (MDE), and the other applied the Calgary Depression Scale for Schizophrenia, the Positive and Negative Syndrome Scale, and the Extrapyramidal Symptoms Rating Scale. Thirteen patients (16.3%) met criteria for MDE. All of them met the DSM-IV PDDS research criteria, but only two patients matched the ICD 10 PSD criteria, which require that the episode occurred in the 12 months after the last psychotic episode. There was no significant difference in the incidence of depressive episodes within 12 months after an acute psychotic episode and outside this time period. The data suggest that depressive episodes in schizophrenia are not restricted to the first year following the psychotic episode. Useful criteria for depressive episodes in schizophrenia should avoid a temporal relation with the psychotic episode. PMID- 12581821 TI - Mood regulation in seasonal affective disorder patients and healthy controls studied in forced desynchrony. AB - In healthy subjects, both the duration of wakefulness and the circadian pacemaker have been demonstrated to be involved in the regulation of mood. Some features of affective disorders suggest that these two factors also play a role in the dysregulation of mood. In particular, disturbances of the circadian pacemaker have been proposed to be a pathogenetic factor in Seasonal Affective Disorder, winter type (SAD). This report presents a test of this proposition. To this end seven SAD patients and matched controls were subjected to a 120-h forced desynchrony protocol, in which they were exposed to six 20-h days. This protocol enables us to discriminate the extent to which the course of mood is determined by the imposed 20-h sleep-wake cycle from the influence of the circadian pacemaker on that course. Patients participated during a depressive episode, after recovery upon light therapy and in summer. Controls were studied in winter and in summer. Between SAD patients and controls no significant differences were observed in the period length nor in the timing of the endogenous circadian temperature minimum. In both groups, sleep-wake cycle- and pacemaker-related components were observed in the variations of mood, which were not significantly different between conditions. PMID- 12581822 TI - Measuring seasonality: psychometric properties of the Seasonal Pattern Assessment Questionnaire and the Inventory for Seasonal Variation. AB - There is a general consensus that seasonal changes in mood, behavior and vegetative functions vary in magnitude across the general population. Studying the dimension of seasonality requires reliable and valid measures. This study examined and compared the psychometric properties of the Seasonal Pattern Assessment Questionnaire (SPAQ) and the Inventory for Seasonal Variation (ISV) in 148 college students; 73 also completed a retest 2 months later. Results indicated good psychometric properties for both scales in terms of score distributions, test-retest reliability, internal consistency, factor structure and item-latent trait relationships using item response theory. Given the long history of its use and some small psychometric and simplicity advantages for the SPAQ, its continued use as a dimensional measure of seasonality is indicated. PMID- 12581823 TI - Elevated blood superoxide dismutase in neuroleptic-free schizophrenia: association with positive symptoms. AB - Blood levels of superoxide dismutase (SOD), measured by radioimmunometric assay, were compared in 68 patients with chronic schizophrenia and 50 normal control subjects. Psychopathology in the patients was assessed with the Brief Psychiatric Rating Scale (BPRS), the Scale for the Assessment of Positive Symptoms (SAPS), and the Scale for the Assessment of Negative Symptoms. Blood SOD levels were significantly elevated in schizophrenia compared with control values. SOD levels showed a positive relationship with the BRPS and the SAPS total score in patients. PMID- 12581824 TI - Stability over time of patients' attitudes toward ECT. AB - This study examined the stability of patients' attitudes toward electroconvulsive therapy (ECT). Surveys were administered to 64 study participants at 2 and 4 weeks post treatment. The survey responses were highly significantly correlated and not significantly different, which suggests that attitudes toward ECT are stable during this time. PMID- 12581825 TI - The effects of vigabatrin on type II spike wave discharges in rats. AB - The antiepileptic drug vigabatrin increases GABA concentrations by inhibiting GABA transaminase. The effects of vigabatrin on type II spike wave discharges (SWDs) in the electroencephalogram of ACI rats were studied in order to learn more about the effects of altering GABA concentration on SWDs. The incidence of type II SWDs increased after vigabatrin (60/h) as compared to saline treatment (3.7/h). This effect appeared with a halftime of 100 min. The duration of type II SWDs increased after vigabatrin (1.52 s) as compared to saline treatment (1.04 s), but the peak-frequency of the type II SWDs decreased after vigabatrin (5.6 Hz) as compared to saline treatment (7.5 Hz). Thus, vigabatrin alters the type II SWD morphology. These results are in agreement with predictions of Destexhe's theoretical model, modulating both GABA(A) and GABA(B) conductances. PMID- 12581826 TI - The ATP-mediated fast current of rat dorsal root ganglion neurons is a novel effector for GABA(B) receptor activation. AB - Because gamma-aminobutyric acid(B) (GABA(B)) agonists produce strong antinociception, the present study analyzed if GABA(B) receptors might operate through depression of P2X(3) receptors responsible for fast adenosine triphosphate (ATP) currents involved in transmitting pain. On rat dorsal root ganglion (DRG) nociceptive neurons, inward currents induced by ATP were inhibited after 2 s or 60 s GABA application and unaffected after 10 s application. SKF 97541 or baclofen, potent GABA(B) agonists, mimicked only the late inhibition of ATP currents. The effect of SKF-97541 or GABA was observed even after their transient application prior to ATP. The GABA(B) antagonist CGP-52432 blocked the action of SKF-97541, suggesting a GABA(B) receptor-mediated mechanism (the GABA(A) antagonist picrotoxin was ineffective). It is suggested that, on nociceptive DRG neurons, GABA produced slow inhibition of P2X(3) receptors via metabotropic GABA(B) receptors. PMID- 12581827 TI - Fibrin is a regulator of Schwann cell migration after sciatic nerve injury in mice. AB - Fibrin, derived from the blood protein fibrinogen, is deposited in the sciatic nerve after injury and retards functional regeneration. Since Schwann cell migration is critical for remyelination of injured nerves, we investigated the effects of fibrin in this process. In vivo experiments showed that fibrin co localizes with fibronectin deposition in the injured sciatic nerve. In vitro migration assays demonstrated that fibrin alone is not a permissive substrate for Schwann cell migration. Furthermore, migration assays of Schwann cells on mixed fibrin/fibronectin substrates showed that fibrin has a dose dependent inhibitory effect on Schwann cell migration on fibronectin. Our results show that fibrin, deposited in the sciatic nerve after injury, changes the composition of the extracellular matrix and inhibits Schwann cell migration. This negative effect of fibrin should be considered in the therapeutic application of biomaterials based on fibrin matrices. PMID- 12581828 TI - Topography of the auditory evoked potential in humans reflects differences between vowels embedded in pseudo-words. AB - To study the processing of vowels embedded in more complex linguistic structures, we compared cortical responses for pseudo-words. Auditory evoked potentials were recorded in 11 right-handed females using a passive oddball paradigm, with /pemu/ and /pomu/ as standard stimuli, differing only with respect to the first syllable. Topographic differences in the N100 were observed between the standards: /pemu/ had larger amplitudes than /pomu/ at more posterior electrode sites whereas a reverse pattern was found at more anterior positions along the midline. This topographic difference can be explained by different generators for the two stimuli. Different vowels and/or the initial formant transition possibly activate different neural populations in the auditory cortex, also when the vowels are embedded in pseudo-words. PMID- 12581830 TI - Deficient auditory processing in children with Asperger Syndrome, as indexed by event-related potentials. AB - Asperger Syndrome (AS) is characterized by normal language development but deficient understanding and use of the intonation and prosody of speech. While individuals with AS report difficulties in auditory perception, there are no studies addressing auditory processing at the sensory level. In this study, event related potentials (ERP) were recorded for syllables and tones in children with AS and in their control counterparts. Children with AS displayed abnormalities in transient sound-feature encoding, as indexed by the obligatory ERPs, and in sound discrimination, as indexed by the mismatch negativity. These deficits were more severe for the tone stimuli than for the syllables. These results indicate that auditory sensory processing is deficient in children with AS, and that these deficits might be implicated in the perceptual problems encountered by children with AS. PMID- 12581829 TI - Increased levels of tyrosine hydroxylase and glutamic acid decarboxylase in locus coeruleus neurons after rapid eye movement sleep deprivation in rats. AB - Norepinephrine, acetylcholine and GABA levels alter during rapid eye movement (REM) sleep and its deprivation. Increased synthesis of those neurotransmitters is necessary for their sustained release. Hence, in this study, the concentrations of tyrosine hydroxylase (TH), choline acetyl transferase (ChAT) and glutamic acid decarboxylase (GAD), the enzymes responsible for their synthesis, were immunohistochemically estimated within the neurons in locus coeruleus, laterodorsal tegmentum and pedunculopontine tegmentum and medial preoptic area in REM sleep deprived and control rats. It was observed that as compared to controls, deprivation increased TH and GAD significantly in the locus coeruleus only, while in other areas, they remained unchanged. The findings help explaining the mechanism of increase in neurotransmitter levels in the brain after REM sleep deprivation and their significance has been discussed. PMID- 12581831 TI - Matrix metalloproteinase-9 deficiency impairs host defense mechanisms against Streptococcus pneumoniae in a mouse model of bacterial meningitis. AB - Matrix metalloproteinase-9 (MMP-9) appears to contribute to blood-brain barrier damage and neuronal injury in bacterial meningitis. To further explore the function of MMP-9 in meningeal inflammation, we injected 10(4) colony forming units (CFU) of a Streptoccocus pneumoniae type 3 strain into the right forebrain of MMP-9 deficient mice (MMP-9(-/-), n=16) and wild-type controls (129 x B6, n=15). The clinical course of the disease, leukocyte recruitment into the subarachnoid space and bacterial titers in the brain did not differ. Yet, clearance of the bacteria from blood (log CFU/ml 4.7 [3.8/5.4] vs. 3.6 [3.0/4.0]; P=0.005) and spleen homogenates (log CFU/ml 5.3 [4.8/5.5] vs. 4.0 [2.8/4.7]; P=0.01) was reduced in MMP-9 deficient mice. A reduced systemic bacterial clearance of MMP-9(-/-) mice was confirmed in experimental S. pneumoniae peritonitis/sepsis. This implies a compromised systemic, but not intracerebral host response against S. pneumoniae in MMP-9 deficiency. PMID- 12581832 TI - Part and whole perceptual-motor practice of a polyrhythm. AB - An experiment is reported that investigated the effectiveness of receiving the perceptual experience of a bimanual, 2:3 polyrhythm during motor practice of the unimanual parts of the polyrhythm. Thirty-six participants were randomly assigned to one of three practice groups: One group practiced both parts of the 2:3 polyrhythm coincident with both pacing metronome tones (whole practice). Another group practiced each rhythm separately, hearing only the pacing tone for the corresponding rhythm (part practice). A third group also practiced each rhythm separately but heard pacing tones for both rhythms during practice (part/whole practice). Each group performed 25, 40 s learning trials for each rhythm; 900 ms intervals for the left hand, and 600 ms intervals for the right hand (a 2:3 polyrhythm). Transfer tests consisted of continuation tapping of the component rhythms, both unimanually and bimanually. Polyrhythmic structure, but not absolute timing stability, was facilitated when training was conducted in the presence of the whole perceptual experience of the task, even when part of the task was practiced unimanually. PMID- 12581833 TI - On-line feedback control of human visually guided slow ramp tracking: effects of spatial separation of visual cues. AB - Visual feedback control of tracking movements is dependent upon a visual comparison of the guiding target and moving limb positions but the human fovea greatly restricts the area of high acuity vision. The effect of vertically separating the target and movement cues in a slow movement task is investigated. Subjects track a slow constant velocity target in the horizontal plane with wrist flexion controlled cursor movements. The effects of changes in the vertical distance between the two cues upon tracking performance were observed. When both cursors were at the same level, tracking was most accurate but showed significant intermittency around 2 Hz in frequency. Increased separation of cues reduced significantly both accuracy and intermittency; tracking was smoother but less accurate. Thus, feedback control is dependent upon the efficiency of positional comparison and hence becomes less effective as the cue separation increases. These results also support previous studies suggesting each cue makes an equal contribution to visuomotor feedback control, each acting as a reference to the other. PMID- 12581834 TI - Evaluation of indices of skeletal muscle contraction in areas of referred hyperalgesia from an artificial ureteric stone in rats. AB - This study examined indices of skeletal muscle contraction in a rat model of referred muscle hyperalgesia from artificial ureteric calculosis [left oblique muscle (OE) for ipsilateral stone]. In specimens from the left versus right OE of stone-implanted female rats, a significant increase was found in membrane fluidity (P<0.01) and Ca(2+)-ATPase activity (P<0.0001) and a significant decrease in 3H-ryanodine binding (P<0.0001) and in I band length/sarcomere length ratio (contraction index) (P<0.01). The increase in Ca(2+)-ATPase activity was directly and significantly related to the number of rats' ureteral 'crises' (P<0.02). The results indicate a state of contraction in the hyperalgesic muscle, whose extent correlates to the algogenic activity of the ureteral stone. PMID- 12581835 TI - The effects of estrogen and raloxifene treatment on the antioxidant enzymes and nitrite-nitrate levels in brain cortex of ovariectomized rats. AB - Number of studies indicate that the female gonadal hormone estrogen protects women against several neurodegenerative diseases and cerebral ischemia via various mechanisms. The possible protective effects of estrogen are mediated mainly by three ways; the activation of steroid receptors and/or modulation of a neurotransmitter and/or direct antioxidative action. Therefore we aimed to investigate the effects of estradiol and raloxifene on levels of nitric oxide (NO) and antioxidant enzymes in brain cortex of ovariectomized female rats. Ten Sprague-Dawley rats were used as naive controls while 32 rats were ovariectomized at 120-140 days of age. Twelve weeks after ovariectomy: (1). Ovariectomized Placebo group (n=11), was given physiologic saline. (2). Estrogen group (n=10) was given Ethynyl estradiol, 0.1 mg/kg sc. (3). Raloxifene group (n=10) was given raloxifene, 1 mg/kg sc. At the end of the treatment period (8 weeks), rats were decapitated and cortex samples were dissected. Results showed that ovariectomy caused a decrease in total nitrite-nitrate levels. The NO levels of both the estrogen and the raloxifene group were higher than the placebo group. Catalase activities did not show any significant difference between the groups, while superoxide dismutase (SOD) activities were elevated via ovariectomy. Estradiol and Raloxifene treatment had no statistically significant effect on SOD activity. PMID- 12581836 TI - Chronic ethanol exposure differentially regulates NOS1 mRNA levels depending on rat brain area. AB - Several works have suggested a potential role for nitric oxide in alcohol-seeking behavior and we have recently shown that the specific blockade of the expression of the neuronal nitric oxide synthase (NOS1) decreases rat ethanol intake. Our previous results have also shown that chronic ethanol exposure has differential effect on the brain NOS activity depending on rat brain area. In the present study, we examine the effects of chronic administration of ethanol on the NOS1 mRNA levels measured with the competitive reverse transcriptase-polymerase chain reaction technique. Chronic administration of ethanol differentially regulated NOS1-mRNA levels depending on rat brain area. Chronic ethanol exposure had no effect on the NOS1-mRNA levels in frontal cortex, but decreased the NOS1-mRNA levels in hippocampus (P<0.01, 39% decrease) and induced a strong increase in striatum (P<0.01, 92% increase). These effects of ethanol were not affected by 7 nitro indazole (25 mg/kg, i.p. daily for 1 week) treatment. These data further support that NOS1 is regulated by chronic exposure to ethanol and that these effects are related to modifications of mRNA levels. PMID- 12581837 TI - ATP is released from nerve terminals and from activated muscle fibres on stimulation of the rat phrenic nerve. AB - Nerve stimulation increases the concentration of ATP in the synaptic cleft, which can act as a neurotransmitter or as a presynaptic neuromodulator. Using the luciferin-luciferase assay, we observed that the extracellular concentration of ATP increased by 11-26 nM over a basal concentration of 6 nM, in a frequency dependent manner (1-5 Hz), in the adult rat phrenic nerve-hemidiaphragm preparation. This ATP release depends on nerve activity since it was abolished by tetrodotoxin (1 microM) and is strictly dependent on the presence of extracellular calcium. However, more than half of this nerve-evoked release of ATP is derived from activated muscle fibres since the selective post-synaptic nicotinic receptor antagonist, alpha-bungarotoxin (1 microM), inhibited by over 60% the evoked release of ATP. The presently observed post-synaptic release of ATP together with the previously reported lack of post-synaptic effects of ATP and to the ability of ATP to act as a presynaptic modulator open the possibility that ATP may behave as a retrograde messenger at this neuromuscular junction. PMID- 12581838 TI - The volatile anesthetic isoflurane suppresses spontaneous calcium oscillations in vitro in rat hippocampal neurons by activation of adenosine A1 receptors. AB - Primary cultures of rat hippocampal neurons were loaded with the Ca(2+)-indicator fluo-3 and studied with a confocal laser microscope. In Mg(2+)-free medium the cultures showed spontaneous synchronized calcium oscillations. These oscillations derived from excitatory signal transmission by N-methyl-D-aspartate and (+/-) alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate receptors and were modulated by gamma-aminobutyric acid(A) receptors. The oscillations were dose-dependently depressed by adenosine (IC50=2 microM) or by 2-chloro-N6 cyclopentyladenosine a specific adenosine A1 receptor agonist (IC50=40 nM). These effects were reverted by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), a specific adenosine A1 receptor antagonist. The volatile anesthetic isoflurane also depressed these spontaneous calcium oscillations in a dose dependent manner (IC50=0.25 MAC, Minimum Alveolar Concentration). The isoflurane-induced inhibition was partly reversed in 29-38% of the neurons by DPCPX, indicating that the anesthetic activates this receptor possibly by increasing the extracellular concentration of adenosine. PMID- 12581839 TI - Immunohistochemical localization of the delta subspecies of protein kinase C in the trigeminal sensory system of Trimeresurus flavoviridis, an infrared-sensitive snake. AB - We examined the expression of the protein kinase C (PKC) delta subspecies in the trigeminal sensory system of the infrared-sensitive snake Trimeresurus flavoviridis. In the trigeminal ganglion (TG), diffuse low-intensity PKC delta immunoreactivity was found in TG neurons and fibers, while intense reactions were observed mainly in medium-sized neurons, which include most of the infrared sensitive neurons. In the brainstem, intense PKC delta immunoreactivity was present in the intermediate layer of the optic tectum of the midbrain and in the nucleus descendens lateralis n. trigemini of the medulla oblongata; these areas are related to the infrared sensory pathway. In the pit organ (the infrared receptor), PKC delta immunoreactivity was present in terminal nerve masses in the pit membrane. These findings suggest that the PKC delta subspecies is involved in the infrared sensory pathway in the trigeminal sensory system of the infrared sensitive snake. PMID- 12581840 TI - Heterotopias, cortical dysplasias and glioneural tumors participate in cognitive processing in patients with temporal lobe epilepsy. AB - Focal brain lesions such as cortical dysplasia and glioneural tumors generate epileptic activity and thus may be synaptically connected with normal cortex. To test this hypothesis, we compared event-related potentials recorded directly from the medial temporal lobe (MTL) and a dysplastic lesion in eight patients with intractable temporal lobe epilepsy. The P3 component, related to visual target detection, showed different peak latencies in four patients and a larger intralesional amplitude compared to established anterior-MTL-generators in two patients. Semantic processing was identified by the N400 component and showed a different latency in four patients and larger intralesional amplitudes in two patients. These results are compatible with the hypothesis that cortical lesions interact with synaptic pathways related to cognitive functions such as visual target detection, and verbal processing. PMID- 12581841 TI - Expression of the sodium channel transcripts Na(v)1.8 and Na(v)1.9 in injured dorsal root ganglion neurons of interferon-gamma or interferon-gamma receptor deficient mice. AB - Changes in the dorsal root ganglion (DRG) expression of the sodium channels Na(v)1.8 and Na(v)1.9 may contribute to injury-induced hyperexcitability and pain. Interferon (IFN)-gamma receptor -/- mice display a reduced pain-related behavior after nerve injury as compared to wild-type mice (NeuroReport 8 (1997) 1311). To elucidate a possible role for IFN-gamma in the regulation of sodium channels, we have studied the DRG mRNA expression of Na(v)1.8/Na(v)1.9 in IFN gamma- or IFN-gamma receptor-deficient mice. In both types of mice, nerve damage induced a downregulation of Na(v)1.8 as well as Na(v)1.9. The magnitude of this reduction was similar to that observed in wild-type animals. These results indicate that the downregulation of Na(v)1.8/Na(v)1.9 in damaged DRG neurons is not influenced by IFN-gamma. Thus, the reduced pain-related behavior of nerve injured IFN-gamma receptor null mice is not due to differential changes in the regulation of Na(v)1.8/Na(v)1.9 mRNA. PMID- 12581842 TI - Altered protein expression levels of Fas/CD95 and Fas ligand in differentially vulnerable brain areas in rats after global cerebral ischemia. AB - To assess the role of the apoptosis-inducing death receptor Fas/CD95 and Fas Ligand (FasL) after global cerebral ischemia, expression of these proteins was investigated in differentially, i.e. selectively vulnerable brain areas. Following experimentally induced cardiac arrest of 6 min duration, rats were resuscitated. After 3, 6, and 24 h of reperfusion, the thalamus and hippocampus of one hemisphere were analyzed for Fas/CD95 and FasL by immunoblotting and semiquantitative densitometry. Corresponding hemispheres were examined by immunohistochemistry. No significant changes in hippocampal Fas/CD95 expression were revealed in comparison to sham operated animals. In the thalamus, a significant reduction in Fas/CD95 expression was observed after 24 h of reperfusion. FasL expression in the hippocampus had declined after 3 and 6 h, as compared with control animals. In contrast, in the thalamus a significant induction of FasL expression was observed after 3 h. Immunohistochemistry revealed a predominantly neuronal expression of the two proteins. In light of the observed increased expression of FasL in the thalamus, such an induction may lead to significant activation of the Fas/CD95 signaling cascade. Our results suggest for the first time a possible role of the Fas/CD95-FasL system after global cerebral ischemia. PMID- 12581843 TI - A family-based association study of T1945C polymorphism in the proline dehydrogenase gene and schizophrenia in the Chinese population. AB - Previous studies have reported genetic linkage evidence for a candidate gene of schizophrenia on chromosome 22q11 but no genes in this region have been really confirmed to be involved in the etiology of schizophrenia so far. Very recently, the proline dehydrogenase gene (PRODH), located in the most centromeric part of the 22q11 microdeletion region, has been reported to be strongly associated with schizophrenia from three sets of independent samples and the most significant evidence for association was derived from a single nucleotide polymorphism PRODH*1945(T/C). We genotyped this polymorphism in 166 Chinese family trios with schizophrenia from East China. No evidence for preferential transmission of the PRODH*1945 alleles from parents to affected offsprings was found using either Transmission Disequilibrium Test (P=0.4) or Haplotype-based Haplotype Relative Risk analysis (P=0.35). Our results suggest that the 1945(T/C) polymorphism of the proline dehydrogenase gene is unlikely to play a major role in the susceptibility to schizophrenia in the Chinese population. PMID- 12581844 TI - Model of retinal surface area and neuron distribution in the avian eye. AB - Changes in retinal neuron distribution may reflect normal or pathological changes in retinal function. The quantitative study of retinal neurons provides a better understanding of anomalous mechanisms, such as those controlling ocular growth and refractive error development in experimental animals. We developed a method to facilitate the quantitative analysis of amacrine neuron populations in wholemount chick retinae, since the domestic chicken is used extensively as an animal model in myopia studies. This method involved automated cell counting from confocal microscopic images and mathematical estimation of total cell numbers based on image cell density and retinal surface areas. Cell densities and cell counts were obtained from immunohistochemically-labeled amacrine neuron populations, using derived formulae to calculate retinal surface area based on vitreous chamber depth, equatorial width, ora serrata diameter and scleral thickness. Normalized total cell counts in each eye were compared, rather than cell densities, since changes in eye growth can affect cell densities. We also compared neuron distribution in central versus peripheral portions of the retina. This is an alternative technique for retinal analysis that supplements traditional anatomical cell counting methods, allowing higher numbers of specimens to be rapidly analyzed. PMID- 12581845 TI - The establishment of a reliable cytotoxic system with SK-N-SH neuroblastoma cell culture. AB - A reliable in vitro cytotoxic system is essential in neurocytotoxic and neuroprotective research. The present study examined four cytotoxic insults with the SK-N-SH human neuroblastoma cell line. These were beta-amyloid protein (Abeta), 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), high density culture, and serum deprivation induced neuronal death. These insults induced significant reduction in cell numbers after 96 h culture, in a concentration dependent manner. Among all the insults, MPTP, serum deprivation, and high density culture induced apoptosis after 96 h, while Abeta presumably induced necrotic neuronal death since apoptosis was not detectable. The p38 MAP kinase inhibitor, SB203580 (1 microM), and the PKC inhibitor, chelerythrine (5 microM) successfully inhibited the loss in viability caused by Abeta and the high density culture, respectively. Other kinase inhibitors, including the non-specific protein kinase inhibitor, H7, the PKA inhibitor 14-22 Amide, the PKG inhibitor, KT5823, and the protein tyrosine kinase inhibitor, AG18 had no effect on any of the four cytotoxic models. This system allows the study of neuroprotection under conditions where the different pathways and mechanisms of the neurons can be considered within one cellular system, removing variations which may be due to different cell type studied. The present studies describe an effective model system for screening potential neuroprotective agents. PMID- 12581846 TI - Three-dimensional load analysis of indentation stimulators. AB - The properties of three types of indentation stimulators: filament indenters, a spring gauge indenter, and an electromechanical stimulator, were evaluated by actuating each device against a 3-direction load cell. The on-axis loads produced by filament indenters were independent of the degree of buckling. The off-axis loads produced by a filament increased with the degree of buckling of the filament, and had magnitudes that were up to 25% of the on-axis load. Repeated applications of a filament produced loads whose magnitudes were very consistent. Offsetting the handle of a filament indenter altered the magnitude and the direction of the off-axis load. Because these factors were poorly controlled in hand held trials, off-axis loads were more variable in hand held trials. The spring gauge stimulator was, when hand held, very susceptible to creating large off-axis loads. When the off-axis loads were large, the spring gauge indicator underestimated the true on-axis loads. The electromechanical stimulator produced loads with a very high degree of consistency and could be arranged so that it produced very small off-axis loads. PMID- 12581847 TI - Optimizing primer--probe design for fluorescent PCR. AB - TaqMan, a variation of fluorescent PCR, is a powerful tool for gene expression and polymorphism studies. Here we describe the design and evaluation of 27 new TaqMan primer-probe sets for rat genes that play a key role in neural signaling. These newly designed and synthesized probes were tested and then used for quantification of RNA isolated from rat brain. The usual length of common TaqMan probes is 25 bases or less. In these studies we constructed probes with lengths of 25-39 bases to span exon-exon junctions of nucleic acids to avoid the influence of DNA contamination upon the RNA quantification. The specific sequences at these positions required probes of these lengths to optimize hybridization. We found that the relocation of the quencher from the traditional 3' position to an internal one increases the sensitivity of probe up to 30 fold. Substitution of 6-carboxyfluorescein with Alexa Fluor 488 as fluorophore and TAMRA with non-fluorescent quencher dabcyl was also investigated. We also describe the evaluation of part of a newly designed set of 27 TaqMan primer probes for the measurement of differences in gene expression levels in samples from the caudate putamen region of rat brain after 'binge' paradigm cocaine administration. Cocaine-induced alterations in expression of c-fos and preprodynorphin mRNAs measured by TaqMan were confirmed by ribonuclease protection assay. PMID- 12581849 TI - Transfer of FGF-2 via HSV-1-based amplicon vectors promotes efficient formation of neurons from embryonic stem cells. AB - The manipulation of embryonic stem (ES) cells has a great potential to create cell populations useful for tissue replacement and regeneration of organs. The acquirement of specific cell fates and differentiation into specialized cell types may be obtained by the application of specific growth factors. Fibroblast growth factor 2 (FGF-2) has been used for the formation of ES cell-derived CNS stem cells. In order to achieve high efficiency of gene delivery into stem cells, the use of viral vectors is presently without alternative. The aim of this study was to examine, for the first time, the ability of herpes simplex virus type-1 (HSV-1)-based amplicon vectors to mediate gene transfer into mouse ES cells and neural progenitors derived from these cells. Amplicon vectors based on HSV-1 have been shown to be a versatile tool to introduce genes into a variety of cell types and to direct their physiological behavior. Here, we show that ES cells and their derivatives can be efficiently transduced and manipulated via these vectors. Amplicon-mediated transfer of FGF-2 into ES cell-derived CNS progenitors leads to the amplification and subsequent differentiation of these precursors into neurons. Therefore, amplicon vectors may now be used to study the effects of other potential factors influencing the cell fate or differentiation of ES cells and their derivatives. PMID- 12581848 TI - High-throughput confirmation of differential display PCR results using reverse Northern blotting. AB - Nylon filter arrays spotted with differential display PCR (DD-PCR) clones and hybridized with radiolabeled cRNA generated from the source RNA pool (reverse Northern blot) provide a high-throughput means to screen clones for artifacts. Reverse Northern blots also confirm differential gene expression in parallel and require modest quantities of the source RNA pool. We describe a strategy to screen multiple candidates from DD-PCR by high-throughput ligation and transformation, followed by reverse Northern blotting. Purification of re amplified DD-PCR clones and fabrication of nylon arrays was facilitated by a batch-processing protocol using the widely available Biomek laboratory robot and Bioworks scripts (available from the authors). A strategy to screen out DD-PCR product artifacts of an inappropriate size was also employed. Using these approaches, we identified several mRNAs that are differentially expressed in response to venlafaxine, fluoxetine or desipramine antidepressant treatment in rat C6 glioma cell lines and are candidates for full length clone isolation using 5'-RACE. Such an approach provides a rapid means to eliminate the high percentage of false positive clones from DD-PCR and enables independent confirmation of differential gene expression patterns generated by various experimental conditions. PMID- 12581850 TI - Visualization of the dendritic arbor of neurons in intact 500 microm thick brain slices. AB - Characterizing the structure and electrophysiological properties of single neurons is essential for understanding how individual cells contribute to the function of neuronal networks. Following intra-cellular recording from neurons in acute brain slices, the structure of the recorded cell has typically been examined by serial sectioning of the tissue slice and then reconstructing the neuron of interest; a labor-intensive and time-consuming process. Here, we have adapted a whole-mount immunohistochemical technique and used it to visualize the dendritic arbor of individual neurons in sections of adult CNS tissue up to 500 microm thick. Permeabilization of the slice and extensive washing allow histochemical reagents to penetrate and be washed from the section, producing limited background staining. Using this method, the cell within the slice can be sectioned optically and reconstructed using the optical sections. We present images of the dendritic trees of neurons in 500 microm thick slices of adult rat entorhinal cortex and hippocampus, labeled either immunohistochemically, or by biocytin injection following whole-cell patch clamp or sharp electrode recordings. The resolution obtained is sufficient to visualize dendritic spines deep within the section. The method is free from artifacts associated with cutting serial sections and is broadly applicable to tasks that require visualization of the fine structure of individual cells in thick slices of CNS tissue. PMID- 12581851 TI - Analysis of EEG records in an epileptic patient using wavelet transform. AB - About 1% of the people in the world suffer from epilepsy and 30% of epileptics are not helped by medication. Careful analyses of the electroencephalograph (EEG) records can provide valuable insight and improved understanding of the mechanisms causing epileptic disorders. Wavelet transform is particularly effective for representing various aspects of non-stationary signals such as trends, discontinuities, and repeated patterns where other signal processing approaches fail or are not as effective. In this research, discrete Daubechies and harmonic wavelets are investigated for analysis of epileptic EEG records. Wavelet transform is used to analyze and characterize epileptiform discharges in the form of 3-Hz spike and wave complex in patients with absence seizure. Through wavelet decomposition of the EEG records, transient features are accurately captured and localized in both time and frequency context. The capability of this mathematical microscope to analyze different scales of neural rhythms is shown to be a powerful tool for investigating small-scale oscillations of the brain signals. Wavelet analyses of EEGs obtained from a population of patients can potentially suggest the physiological processes undergoing in the brain in epilepsy onset. A better understanding of the dynamics of the human brain through EEG analysis can be obtained through further analysis of such EEG records. PMID- 12581852 TI - A murine model of subarachnoid hemorrhage-induced cerebral vasospasm. AB - Cerebral vasospasm remains a major cause of morbidity and mortality after subarachnoid hemorrhage (SAH). The availability of a mouse model of SAH that is simple, replicable and has low mortality would provide a powerful approach for understanding cellular and molecular mechanisms contributing to post-SAH pathologies. The present study characterizes a mouse model of experimental SAH, which produces consistent constriction of large cerebral arteries. Adult mice received injections of autologous blood into the cisterna magna, and the diameters of large intracranial vessels were measured 1 h to 7 days post-SAH. A diffuse blood clot was evident in both the anterior and posterior circulations after SAH. Vascular wall thickening, lumenal narrowing and corrugation of the internal elastic lamina were observed. Both acute (6-12 h) and delayed (1-3 days) phases of vasoconstriction occurred after SAH. Overall mortality was only 3%. A reproducible, low mortality model of SAH-induced cerebral vasospasm in mice is described. This mouse model should facilitate the delineation of cellular and molecular mechanisms of SAH-induced pathologies because of the widespread availability of various technologies for this species (e.g. genetically-altered animals and gene expression arrays). This model also represents a replicable and inexpensive approach for screening therapeutic candidates. PMID- 12581853 TI - A method utilizing differential culture and comparative RT-PCR for determining RNA expression in superior cervical ganglion neurones. AB - In order to assess the neuronal expression of caspase mRNA in primary cultures of rat superior cervical ganglion (SCG) neurones a method of differential cell purification and comparative RT-PCR was devised. SCG primary cultures generally contain variable percentages of non-neuronal contaminants, which influence RT-PCR results. We optimised a neuronal purification method, allowing the preparation of both highly purified neuronal cultures and mixed cultures, enriched in non neuronal contaminants. These two sets of cells were cultured in parallel and subsequently analysed by RT-PCR. The use of cell type specific oligonucleotides allowed evaluation of the relative distribution of neuronal (neurofilament) and non-neuronal transcripts in the two cultures. In parallel, specific oligonucleotides were used to detect the mRNA levels of caspase family members. The partition of neurofilament transcript between pure and mixed cultures was found to be statistically different from the partition of the non-neuronal markers. Therefore statistical difference from the partition of non-neuronal markers was taken as evidence for expression in neurones. We show that caspase-2, -3, -6, -7 and -9 transcripts are expressed in SCG neurones whereas caspase-1 is probably absent. Furthermore, none of these transcripts are upregulated during neuronal death induced by nerve growth factor withdrawal. This method could be applied to the analysis of other transcripts in SCG and other primary neuronal cultures containing significant percentages of contaminant cell types. PMID- 12581854 TI - A device for automated control of pipette internal pressure for patch-clamp recording. AB - Formation of a high-resistance seal between the tip of a glass recording pipette and the membrane of the recorded cell is the crucial step in patch clamping, or whole cell recording with patch pipettes. Formation of the seal, and subsequent rupture of the membrane for whole cell recording, requires a specific sequence of changes in pipette internal hydrostatic pressure. Generating this sequence of pressure changes adds to the complexity of setting up, gaining proficiency, and performing experiments. Automation of routine pipette pressure manipulations would simplify seal formation, and benefit productivity. Here we describe a device that automates control of patch pipette internal pressure. Solenoid valves sequentially operated by manual switching, or external electronic control, automatically provide the necessary sequence of connections to the pipette interior. This greatly simplifies the operations performed to obtain membrane seals and whole cell recordings and improves standardization and reproducibility in patch recording. PMID- 12581855 TI - Zyxin and paxillin proteins: focal adhesion plaque LIM domain proteins go nuclear. AB - Zyxin and paxillin are the prototypes of two related subfamilies of LIM domain proteins that are localized primarily at focal adhesion plaques. However, recent work has shown that zyxin/paxillin family proteins also shuttle through the nucleus. These proteins may enter the nucleus by association with other proteins, but are exported from the nucleus by means of intrinsic leucine-rich nuclear export sequences. Zyxin/paxillin proteins may regulate gene transcription by interaction with transcription factors. In some cases, misregulation of nuclear functions of zyxin/paxillin proteins appear to be associated with pathogenic effects. PMID- 12581856 TI - Rho GTPases and spermatogenesis. AB - Rho GTPases, such as Rho, Rac and Cdc42, are known to regulate many cellular processes including cell movement and cell adhesion. While the cellular events of germ cell movement are crucial to spermatogenesis since developing germ cells must migrate progressively from the basal to the adluminal compartment but remain attached to the seminiferous epithelium, the physiological significance of Rho GTPases in spermatogenesis remains largely unexplored. This paper reviews some recent findings on Rho GTPases in the field with emphasis on the studies in the testis, upon which future studies can be designed to delineate the role of Rho GTPases in spermatogenesis. PMID- 12581857 TI - Low-energy laser irradiation enhances de novo protein synthesis via its effects on translation-regulatory proteins in skeletal muscle myoblasts. AB - Low-energy laser irradiation (LELI) drives quiescent skeletal muscle satellite cells into the cell cycle and enhances their proliferation, thereby promoting skeletal muscle regeneration. Ongoing protein synthesis is a prerequisite for these processes. Here, we studied the signaling pathways involved in the LELI regulation of protein synthesis. High levels of labeled [35S]methionine incorporation were detected in LELI cells as early as 20 min after irradiation, suggesting translation of pre-existing mRNAs. Induced levels of protein synthesis were detected up until 8 h after LELI implying a role for LELI in de novo protein synthesis. Elevated levels of cyclin D1, associated with augmented phosphorylation of the eukaryotic initiation factor 4E (eIF4E) and its inhibitory binding protein PHAS-I, suggested the involvement of LELI in the initiation steps of protein translation. In the presence of the MEK inhibitor, PD98059, eIF4E phosphorylation was abolished and levels of cyclin D1 were dramatically reduced. The LELI-induced PHAS-I phosphorylation was abolished after preincubation with the PI3K inhibitor, Wortmannin. Concomitantly, LELI enhanced Akt phosphorylation, which was attenuated in the presence of Wortmannin. Taken together, these results suggest that LELI induces protein translation via the PI3K/Akt and Ras/Raf/ERK pathways. PMID- 12581858 TI - Characterisation of PDZ-GEFs, a family of guanine nucleotide exchange factors specific for Rap1 and Rap2. AB - PDZ-GEF1 (RA-GEF/nRapGEP/CNrasGEF) is a guanine nucleotide exchange factor (GEF) characterised by the presence of a PSD-95/DlgA/ZO-1 (PDZ) domain, a Ras association (RA) domain and a region related to a cyclic nucleotide binding domain (RCBD). These domains are in addition to a Ras exchange motif (REM) and GEF domain characteristic for GEFs for Ras-like small GTPases. PDZ-GEF1 efficiently exchanges nucleotides of both Rap1 and Rap2, but has also been implicated in mediating cAMP-induced Ras activation through binding of cAMP to the RCBD. Here we describe a new family member, PDZ-GEF2, of which we isolated two splice variants (PDZ-GEF2A and 2B). PDZ-GEF2 contains, in addition to the domains characteristic for PDZ-GEF1, a second, less conserved RCBD at the N terminus. PDZ-GEF2 is also specific for both Rap1 and Rap2. We further investigated the possibility that PDZ-GEF2, like PDZ-GEF1, is a cAMP-responsive GEF for Ras. However, in contrast to previous results, we did not find any effect of either PDZ-GEF1 or PDZ-GEF2 on Ras in the absence or presence of cAMP. Moreover, affinity measurements by isothermic calorimetry showed that the RCBD of PDZ-GEF1 does not bind cAMP with a physiologically relevant affinity. We conclude that both PDZ-GEF1 and 2 are specific for Rap1 and Rap2 and unresponsive to cAMP and various other nucleotides. PMID- 12581859 TI - Molecular properties of endogenous RFamide-related peptide-3 and its interaction with receptors. AB - Based on database searches of DNA sequences, we previously reported a gene encoding peptides possessing Arg-Phe-NH(2) (RFamide) at their C termini. This gene, RFamide-related peptide (RFRP), was expected to encode several different peptides (i.e., RFRP-1, -2, and -3). In the present study, we purified endogenous RFRP-3 from bovine hypothalamus, and demonstrated that it consisted of 28 amino acid residues. After constructing a sandwich enzyme immunoassay for RFRP-3, we analyzed the tissue distribution of endogenous RFRP-3 in rats and found its concentration to be highest in the hypothalamus. In binding assays, [125I] labeled RFRP-3 bound to OT7T022 with high affinity, but its binding affinity to HLWAR77 was low. On the other hand, [125I]-labeled neuropeptide FF (NPFF) bound to both OT7T022 and HLWAR77 with high affinity. By serial deletion in the N terminal portions of RFRP-3 and NPFF, we found that four C-terminal amino acid residues (i.e., PQRFamide), which were common between the two peptides, comprised a core sequence responsible for binding with the receptors, whereas three amino acid residues (i.e., PNL in RFRP-3 and LFQ in NPFF) added to the N terminus of PQRFamide played crucial roles in the agonistic activities of RFRP-3 and NPFF for OT7T022 and HLWAR77, respectively. PMID- 12581860 TI - gamma-Irradiation-induced DNA damage enhances NO production via NF-kappaB activation in RAW264.7 cells. AB - We investigated the mechanism of augmentation of nitric oxide (NO) production in the murine macrophage cell line RAW264.7 after gamma-irradiation. The cells treated with interferon-gamma (IFN-gamma) or lipopolysaccharide (LPS) showed enhanced NO production by gamma-irradiation in a dose-dependent manner, accompanying the induction of inducible nitric oxide synthase (iNOS) expression. Nuclear factor kappa B (NF-kappaB) activation was induced 1 h after gamma irradiation dose-dependently, which was detected by the degradation of I-kappaB. Inhibitors of I-kappaB degradation, MG132 and N(alpha)-p-tosyl-L-lysine chloromethyl ketone (TLCK), suppressed the further increase by gamma-irradiation in IFN-gamma-induced NO production, showing that gamma-irradiation induced NO production via NF-kappaB activation. Although NF-kappaB is known to be a redox sensitive transcription factor, the antioxidant agents N-acetyl-cysteine (NAC) and 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (trolox) showed no suppression and treatment with H(2)O(2) showed only slight enhancement of IFN gamma-induced NO production. The DNA damaging agents camptothecin and etoposide enhanced IFN-gamma-induced NO production and showed I-kappaB degradation, indicating that the increase in NO production was due to direct DNA damage. Furthermore, 3-aminobenzamide (3AB) and benzamide, inhibitors of poly (ADP ribose) polymerase (PARP) that are activated upon recognition of DNA strand breaks, suppressed the further increase by gamma-irradiation in IFN-gamma-induced NO production and the I-kappaB degradation by gamma-irradiation. We concluded that (1) the increase in NO production was due to direct DNA damage by gamma irradiation, and that (2) PARP activation through DNA damage induced NF-kappaB activation, leading to iNOS expression and NO production. PMID- 12581861 TI - Effect of ageing in the early biochemical signals elicited by PTH in intestinal cells. AB - In previous work, we have demonstrated that rPTH(1-34) increases cytoplasmic calcium concentration ([Ca(2+)](i)) in isolated rat enterocytes. In the present study, we have identified the sources of PTH-mediated increase in [Ca(2+)](I) and the implication of Ca(2+) on hormone early signals in enterocytes isolated from young (3-month-old) and aged (24-month-old) rats. In young enterocytes, PTH raised [Ca(2+)](i) in a dose-dependent manner (1 pM-100 nM). In cells from aged rats, hormone concentrations higher than physiological (>/=1 nM) were required to observe significant increases in [Ca(2+)](i). Phospholipase C (PLC) inhibitors blocked the initial acute elevation of the [Ca(2+)](i) biphasic response to PTH of young enterocytes while in old cells, no effects were observed. The voltage dependent calcium-channel blocker (VDCC), nitrendipine, suppressed PTH-dependent changes of the sustained [Ca(2+)](i) phase in young and aged animals. In this study, we analysed, for the first time, alterations in phosphatidylinositol 3 kinase (PI3K) activity and response to PTH in rat enterocytes with ageing. Basal PI3K activity was significantly modified by ageing. Acute treatment with 10(-8) M PTH increased enzyme activity, with a maximun at 2 min (+3-fold) in young rats and only elevated by less than 1-fold basal PI3K activity in aged animals. Hormone-induced tyrosine phosphorylation of p85alpha, the regulatory subunit of PI3K, as well as the phosphorylation on Thr(308) of its downstream effector Akt/PKB was evident in enterocytes from 3-month-old rats, whereas it was greatly reduced in the cells from 24-month-old animals. Intracellular Ca(2+) chelation (BAPTA-AM, 5 microM) affected the tyrosine phosphorylation of p85alpha and inhibited PTH-dependent PI3K activation by 75% in young rats and completely abolished the enzyme activity in aged animals, demonstrating that Ca(2+) is required for full activation of PI3K in enterocytes stimulated with PTH. The Thr phosphorylation of PI3K downeffector, Akt/PKB, was also fully dependent on Ca(2+). Taken together, these results suggest that PTH regulation of enterocyte [Ca(2+)](i) involves Ca(2+) mobilization from IP(3)-sensitive stores and the influx of the cation from the extracellular milieu, the former pathway being blunted during ageing. The data also indicates a positive role for intracellular calcium in one of the early signals of PTH in rat enterocytes, the activation of PI3K, and that hormone regulation of PI3K activity and Akt/PKB phosphorylation on Thr(308) is impaired with ageing. PMID- 12581862 TI - Caspase-mediated apoptosis in sponges: cloning and function of the phylogenetic oldest apoptotic proteases from Metazoa. AB - Sponges (phylum Porifera) represent the phylogenetically oldest metazoan phylum. These animals have complex cell adhesion and powerful immune systems which allow the formation of a distinct body plan. Consequently, an apoptotic machinery has to be predicted that allows sponges to eliminate unwanted cells accumulating during development. With the marine sponge Geodia cydonium, it is shown that allografts of these animals undergo apoptosis as demonstrated by apoptotic DNA fragmentation. Extracts from allografts contain an enzymic activity characteristic for caspases; as substrate to determine the cleavage activity, Ac DEVD-AMC was applied. cDNAs encoding predicted caspase-3-related proteins were isolated; they comprise the characteristic structure known from caspases of other metazoan phyla. The two cDNAs are assumed to originate from one gene by alternative splicing; the longer form comprises a caspase recruitment domain (CARD), whereas the shorter one is missing CARD. The expression of sponge caspase genes is up-regulated during allograft rejection. In vivo incubation experiments with Ac-DEVD-CHO (a caspase-3 inhibitor) showed a reduction of apoptotic DNA fragmentation, whereas Ac-LEHD-CHO (an inhibitor of caspase-9) caused no effect. It is concluded, that for the establishment of the metazoan body plan, both the adhesion molecules and the apoptotic molecules (described here) were essential prerequisites. PMID- 12581863 TI - Role of calcium in phototoxicity of 2-butylamino-2-demethoxy-hypocrellin A to human gastric cancer MGC-803 cells. AB - After incubation with 2-butylamino-2-demethoxy-hypocrellin A (2-BA-2-DMHA), photodynamically induced change in the cytoplasmic free calcium concentration ([Ca(2+)](i)) and its effect on cell damage were investigated in human gastric cancer (MGC-803). Fluorescence spectrophotometry measurement indicated that the photosensitization of MGC-803 by 2-BA-2-DMHA caused an increase in intracellular calcium [Ca(2+)](i), and this increase in [Ca(2+)](i) showed a dependence on the concentration of 2-BA-2-DMHA, light dose and extracellular [Ca(2+)](e). This phenomenon of intracellular calcium accumulation was further confirmed by using laser scanning confocal microscopy (LSCM). Furthermore, the results from MTT assay and flow cytometry analysis suggested that chelation of extracellular calcium by EGTA or intracellular calcium by BAPTA could inhibit photodynamically induced cell killing, while increase of [Ca(2+)](i) by thapsigargin (TG), a highly specific inhibitor of the Ca(2+)-ATPase, or by A23187, a calcium ionophore could enhance this action. Meanwhile, the nucleus morphology was also investigated by fluorescence microscopy. The results indicated that the increase in intracellular Ca(2+) concentration was responsible for 2-BA-2-DMHA photodynamically induced damage to MGC-803. PMID- 12581864 TI - Distinct regulation of glucose transport and GLUT1/GLUT3 transporters by glucose deprivation and IGF-I in chromaffin cells. AB - Effects of prolonged metabolic (glucose deprivation) and hormonal [insulin-like growth factor I (IGF-I)] challenge on regulation of glucose transporter (GLUT) expression, glucose transport rate and possible signaling pathways involved were studied in the neuroendocrine chromaffin cell. The results show that bovine chromaffin cells express both GLUT1 and GLUT3. Glucose deprivation and IGF-I activation led to an elevation of GLUT1 and GLUT3 mRNA, the strongest effect being that of IGF-I on GLUT3 mRNA. Both types of stimulus increased the GLUT1 protein content in a cycloheximide (CHX)-sensitive manner, and the glucose transport rate was elevated by 3- to 4-fold after 48 h under both experimental conditions. IGF-I-induced glucose uptake was totally suppressed by CHX. In contrast, only approximately 50% of transport activation in glucose-deprived cells was sensitive to the protein synthesis inhibitor. Specific inhibitors of mTOR/FRAP and p38 MAPK each partially blocked IGF-I-stimulated glucose transport, but had no effect on transport rate in glucose-deprived cells. The results are consistent with IGF-I-activated transport being completely dependent on new GLUT protein synthesis while the enhanced transport in glucose-deprived cells was partially achieved independent of new synthesis of proteins, suggesting a mechanism relying on preexisting transporters. PMID- 12581865 TI - Interactions between doxorubicin and the human iron regulatory system. AB - Anthracyclines are included in clinical treatments against various malignancies, but severe cardiotoxic side-effects and the development of resistance mechanisms limit their usefulness. Many aspects of the cellular response to anthracyclines remain debated. The status of the main regulator of iron homeostasis, namely the RNA-binding activity of iron regulatory proteins (IRPs), has been assessed herein for two types of human tumor cells and their derived doxorubicin-resistant sublines. IRPs were always fully activated in the latter, whereas only partial activation occurred in the former. Doxorubicin exposure reversibly inactivated IRP1 in small cell lung carcinoma (GLC(4)) and myelogenous leukemia (K562) cell lines, but was without effect in their derived doxorubicin-resistant sublines. In contrast, adding doxorubicin to cytosolic fractions of untreated cells or to purified IRPs led to the irreversible alteration of the RNA-binding activity of IRP1. In these different conditions, interaction between doxorubicin and the iron regulatory system disturbs iron metabolism, and cells having developed a resistance mechanism are tuned to maximize the iron supply. The results reported herein may lead the path toward a better therapeutic management of cancer patients receiving doxorubicin by discriminating between the antiproliferative and cardiotoxic properties of this anthracycline. PMID- 12581866 TI - PKC epsilon -mediated ERK1/2 activation involved in radiation-induced cell death in NIH3T3 cells. AB - Protein kinase C (PKC) isoforms play distinct roles in cellular functions. We have previously shown that ionizing radiation activates PKC isoforms (alpha, delta, epsilon, and zeta), however, isoform-specific sensitivities to radiation and its exact mechanisms in radiation mediated signal transduction are not fully understood. In this study, we showed that overexpression of PKC isoforms (alpha, delta, epsilon, and zeta) increased radiation-induced cell death in NIH3T3 cells and PKC epsilon overexpression was predominantly responsible. In addition, PKC epsilon overexpression increased ERK1/2 activation without altering other MAP kinases such as p38 MAPK or JNK. Co-transfection of dominant negative PKC epsilon (PKC epsilon -KR) blocked both PKC epsilon -mediated ERK1/2 activation and radiation-induced cell death, while catalytically active PKC epsilon construction augmented these phenomena. When the PKC epsilon overexpressed cells were pretreated with PD98059, MEK inhibitor, radiation-induced cell death was inhibited. Co-transfection of the cells with a mutant of ERK1 or -2 (ERK1-KR or ERK2-KR) also blocked these phenomena, and co-transfection with dominant negative Ras or Raf cDNA revealed that PKC epsilon -mediated ERK1/2 activation was Ras-Raf dependent. In conclusion, PKC epsilon -mediated ERK1/2 activation was responsible for the radiation-induced cell death. PMID- 12581867 TI - Tissue kallikrein is synthesized and secreted by human vascular endothelial cells. AB - The generation of kinins on the surface of vascular endothelium has been postulated in two pathways involving plasma kallikrein and tissue kallikrein; the former pathway has been well documented, but the latter is controversial. To clarify the presence of a kinin-generating system on endothelium, we examined whether human umbilical vein endothelial cells (HUVEC) synthesize and release tissue kallikrein in vitro. Kallikrein-like activity hydrolyzing a peptide Pro Phe-Arg-4-methyl-coumaryl-7-amide was detected in the culture medium of HUVEC and was inhibited by aprotinin but not by soybean trypsin inhibitor. Western blotting of HUVEC medium using anti-human tissue kallikrein antibodies demonstrated the release of tissue kallikrein from HUVEC, and the reverse transcription-polymerase chain reaction (RT-PCR) followed by Southern blotting revealed the expression of tissue kallikrein mRNA in HUVEC. HUVEC metabolically labeled with [35S]methionine released radioactive proteins corresponding to tissue kallikrein. RT-PCR also showed the expression of low-molecular-weight kininogen (L-kininogen) mRNA in HUVEC. The cGMP levels in HUVEC were significantly elevated by the incubation with angiotensin converting enzyme inhibitor, lisinopril, and the elevation was completely inhibited by aprotinin or bradykinin B2-receptor antagonist, FR172357. These results suggest that the endothelial cells continuously release an active form of tissue kallikrein which enables generation of kinins on the vascular endothelium. PMID- 12581868 TI - Tensile stress-dependent collagen XII and fibronectin production by fibroblasts requires separate pathways. AB - The intracellular mechanisms controlling mechano-dependent production of the two extracellular matrix proteins collagen XII and fibronectin were analyzed. Fibroblasts were cultured on either tensed (attached) or released (floating) collagen type-I gels, respectively. Collagen XII and fibronectin production was three- to fivefold higher under tensed than under released conditions. The general inhibitor of tyrosine phosphorylation, genistein (50 microM), and the MAP kinase inhibitor PD98059 (20 microM) selectively reduced collagen XII accumulation by tensed cultures. Addition of PD98059, but not genistein, downregulated tensile stress-induced tyrosine phosphorylation levels of ERK1/2 and focal adhesion kinase. Staurosporine as well as pretreatment with phorbol ester, which constitute means to downregulate classical and novel PKC activity, specifically blocked collagen XII but not fibronectin accumulation in tensed fibroblasts. ERK1/2 phosphorylation levels were not affected by staurosporine treatment. Chronic exposure to the protein kinase C inhibitors bisindolylmaleimide and calphostin C blocked increased production of both fibronectin and collagen XII from cells under tension. The data manifest that the mechano-dependent production of collagen XII and fibronectin requires separate pathways. The FAK-ERK1/2 pathway, a genistein-sensitive tyrosine kinase, and a distinct classical/novel PKC appear selectively required for increased production of collagen XII in cells under tensile stress, whereas fibronectin induction is regulated by a different PKC-dependent pathway. PMID- 12581869 TI - Cytochalasin D disruption of actin filaments in 3T3 cells produces an anti apoptotic response by activating gelatinase A extracellularly and initiating intracellular survival signals. AB - Disruption of actin filaments affects multiple cell functions including motility, signal transduction and cell division, ultimately culminating in cell death. Although this is the usual sequence of events, we have made the interesting observation that disruption of actin filaments by the potent toxin cytochalasin D (Cyto D) causes one cell type, mouse mesangial cells (MMC), to undergo apoptosis, while in another cell type (NIH 3T3), it has the opposite effect, resulting in production of survival signals. The purpose of this study was to investigate the molecular basis for these observed differences. In the present communication, we demonstrate that exposure to Cyto D induces the pro-apoptotic pathways, p38 and stress-activated protein kinase (SAPK)/jun amino-terminal kinase (JNK), in both cell types. However, in 3T3, but not MMC, the extracellular signal regulated kinase (ERK) 1/2 pathway is protected from inhibition following treatment with Cyto D-leading to phosphorylation of Bclxi/Bcl 2-associated death promoter (BAD). Inhibition of Cyto D-induced secretion and activation of gelatinase A in 3T3 cells reverses the production of survival signals by Cyto-D. To investigate this effect further we employed CS-1 cells, a well-characterized melanoma cell line that lacks integrin beta3, and also does not secrete gelatinase A. Co transfection of CS-1 cells with integrin beta3 and a gelatinase A transgene, which enables the cells to secrete constituitively active gelatinase A, enhances CS-1 cell survival signals. Together, our findings suggest that extracellularly activated gelatinase A, through interaction with integrin alphaVbeta3, elicits survival signals mediated through ERK 1/2 that override activation of p38 and SAPK/JNK stress pathways. PMID- 12581870 TI - Serglycin proteoglycan expression and synthesis in embryonic stem cells. AB - The serglycin proteoglycan is expressed in most hematopoietic cells and is packaged into secretory vesicles for constitutive or regulated secretion. We have now shown serglycin mRNA expression in undifferentiated murine embryonic stem (ES) cells and in embryoid bodies, and synthesis and secretion in undifferentiated ES cells. Serglycin was localized to ES cell cytoplasm by immunostaining. Serglycin mRNA is expressed in tal-1((-/-)) ES cells and embryoid bodies; tal-1((-/-)) mice cannot produce hematopoietic cells. Thus, ES serglycin expression is probably not associated with hematopoiesis. Serglycin expression was increased by treatment of ES cells with retinoic acid (RA) and dibutyryl cAMP (dbcAMP). The serglycin core protein obtained from control ES culture medium after chondroitinase digestion appears as a doublet. Only the lower Mr band is present in serglycin secreted from RA-treated and the higher Mr band in RA+dbcAMP treated cells, suggesting that core protein structure is affected by differentiation. PMID- 12581871 TI - Hypoxia up-regulates glyceraldehyde-3-phosphate dehydrogenase in mouse brain capillary endothelial cells: involvement of Na+/Ca2+ exchanger. AB - The molecular regulatory mechanisms and the characterization of glyceraldehyde-3 phosphate dehydrogenase (GAPDH) in hypoxia were studied in a mouse brain capillary endothelial cell line, MBEC4. Activation of GAPDH gene expression by hypoxia was suppressed by an intracellular Ca(2+) chelator and inhibited by a non selective cation channel blocker or a Na(+)/Ca(2+) exchanger (NCX) blocker. Sequencing of reverse transcription-PCR products demonstrated that MBEC4 expressed an mRNA encoding NCX3, which functions even under cellular ATP-depleted conditions, in addition to mRNAs encoding NCX1 and NCX2. The inhibition of Ca(2+)/calmodulin-dependent protein kinases or c-Jun/AP-1 activation caused a significant decrease in the activation of GAPDH mRNA by hypoxia. These results suggest that hypoxia stimulates Ca(2+) influx through non-selective cation channels and causes the reverse operation of the three NCX isoforms, and consequently, increased intracellular Ca(2+) up-regulates GAPDH gene expression through an AP-1-dependent pathway. Furthermore, subcellular fractionation experiments showed that hypoxia increased GAPDH proteins not only in the cytosolic fraction, but also in the nuclear and particulate fractions, in which GAPDH should play no roles in glycolysis. However, the GAPDH activity did not rise in proportion to the increase of GAPDH protein by hypoxia even in the cytosolic fraction. These results suggest that not all hypoxia-induced GAPDH molecules contribute to glycolysis. PMID- 12581872 TI - The novel retinoid AHPN/CD437 induces a rapid but incomplete apoptotic response in human myeloma cells. AB - The synthetic retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (AHPN/CD437) appears to possess an apoptotic activity superior to classical retinoids in vitro as in vivo. Numerous studies have shown that CD437 induced apoptosis is independent of its nuclear receptor activity, suggesting that CD437 might have a unique mechanism of action. The purpose of this study was to compare CD437- and all-trans retinoic acid (atRA)-induced cell death. CD437 provoked a rapid apoptotic phenotype immediately followed by secondary necrosis in RPMI 8226, U266 and L363 human myeloma cell lines. Nuclear apoptotic features were observed upon both CD437 and atRA treatments. In contrast, membrane blebbing and the subsequent formation of apoptotic bodies, a classical apoptotic event, was only observed upon atRA treatment. In addition, CD437, contrary to atRA, was unable to induce tissue transglutaminase (tTG), an intracellular enzyme involved in the formation of cross-linked protein polymers contributing to apoptotic morphological changes. Taken together, these data suggest that CD437 induces rapid but incomplete apoptotic phenotype in human myeloma cells. PMID- 12581873 TI - Marked induction of sterol 27-hydroxylase activity and mRNA levels during differentiation of human cultured monocytes into macrophages. AB - Sterol 27-hydroxylase has been suggested to be involved in an alternative pathway for the elimination of cholesterol from macrophages and early atherosclerotic lesions. We have previously shown that human lung macrophages as well as monocyte derived macrophages have a relatively high activity of sterol 27-hydroxylase (CYP27). This enzyme converts intracellular cholesterol into 27 hydroxycholesterol and cholestenoic acid that flux from cultured cells into the medium. It is shown here that human monocytes have very low CYP27 activity and CYP27 mRNA levels. During differentiation into macrophages, both CYP27 activity and CYP27 mRNA levels increase markedly after 4 days of culture in serum-free medium. Addition of macrophage-colony stimulating factor had no significant effect on the induction and addition of fetal calf serum had an inhibitory effect. Cholesterol synthesis was found to be a critical factor for the production of 27-oxygenated products by the macrophages cultured in serum-free medium. The increased capacity of the differentiated cells to eliminate intracellular cholesterol is of interest and supports the contention that CYP27 is an antiatherogenic factor. PMID- 12581874 TI - Corpus luteum derived copper, zinc-superoxide dismutase serves as a luteinizing hormone-release inhibiting factor in sheep. AB - In the present study, we report the purification and characterization of a polypeptide from the sheep corpus luteum of pregnancy with a potent luteinizing hormone-release inhibiting factor (LH-RIF) bioactivity that stained as a single band in SDS-PAGE with an apparent molecular mass of 16000 Da. The amino acid sequences obtained after sequence analysis of peptides derived from the trypsin digestion of LH-RIF were subjected to a protein data bank search and were found to be identical with regions of sheep copper, zinc-superoxide dismutase (Cu,Zn SOD). The measured mass of LH-RIF (15604.2+/-1.9 Da) was found to be similar to the theoretical mass of sheep Cu,Zn-SOD (15603.5 Da), with a disulfide bond and N acetylated alanine at the N-terminus. The inhibitory action of Cu,Zn-SOD on pulsatile LH secretion would suggest that this antioxidant may play an important role, either independently or in concert with some neurotransmitters, in the neuroendocrine regulation of sheep female reproductive function. PMID- 12581875 TI - Structure and juvenile hormone-mediated regulation of the HMG-CoA reductase gene from the Jeffrey pine beetle, Dendroctonus jeffreyi. AB - In several pine bark beetle species, juvenile hormone (JH) III regulated 3 hydroxy-3-methylglutaryl coenzyme A reductase (HMG-R) gene expression has an important role in monoterpenoid pheromone production in males. We investigated the structure and regulated expression of the HMG-R gene (HMG-R) in the Jeffrey pine beetle, Dendroctonus jeffreyi. cDNA and genomic sequences were recovered using a combination of library screening and PCR. The transcribed portion of the gene spans 9.8 kb and is interrupted by 13 introns. When compared to vertebrate HMG-Rs, the distribution of intron sites suggests a functional role for those in the 5' untranslated region and membrane anchor domains. Northern blots show that topically applied JH III stimulates HMG-R expression up to 30-fold in male D. jeffreyi, compared to untreated insects, in both a dose- and time-dependent manner. There was no increase in expression levels in similarly treated female insects. The expression pattern is consistent with the production of monoterpenoid pheromone components in male D. jeffreyi, and suggests the utility of the system as a new tool for studying the mechanism of JH action. PMID- 12581876 TI - Endotoxin decreases serum IGFBP-3 and liver IGFBP-3 mRNA: comparison between Lewis and Wistar rats. AB - We studied the IGFBP-3 response to endotoxin, in Wistar and Lewis rats. Compared to Wistar rats, Lewis rats have a reduced adrenal and IGF-I response to inflammatory stimuli. Rats received two injections of 1 mg/kg of lipopolysaccharide (LPS) and were killed 4 h after the second injection. LPS decreased serum concentrations of GH in Wistar (P<0.05), but not in Lewis rats. However, serum IGFBP-3 was decreased both in Wistar and in Lewis rats. Furthermore, LPS administration decreased IGFBP-3 gene expression in the liver in both rat strains (P<0.01). Lewis rats had lower serum IGFBP-3 than Wistar rats (P<0.01). This difference could be secondary to the increased IGFBP-3 proteolysis in serum observed in Lewis rats. These data indicate that acute inflammation inhibits serum concentrations of IGFBP-3 by decreasing its synthesis in the liver, rather than increasing its proteolysis. This effect seems to be GH and IGF I independent. PMID- 12581877 TI - Gonadotropin-releasing hormone-induced stimulation of the rat secretogranin II promoter involves activation of CREB. AB - To investigate the events involved in regulation of the secretogranin II (SgII) gene, luciferase reporter constructs were transfected into gonadotrope-derived, alphaT3-1 cells. DNA between -91 and -60 relative to the transcription start site was found to be required for GnRH induced SgII reporter gene activation. This region contains a consensus cAMP response element (CRE) and disruption of this CRE reduced GnRH responsiveness of the SgII promoter. CREB was shown to bind to the SgII CRE and transfection studies with a dominant-negative CREB mutant provided evidence that CREB is required for GnRH responsiveness of the SgII promoter. An expression vector for an inhibitor of the cAMP-dependent protein kinase was found to reduce the ability of cAMP or GnRH to activate the SgII luciferase reporter gene. These studies offer evidence that GnRH-induced activation of the SgII promoter in the alphaT3-1 cell line requires cAMP dependent protein kinase activity and a functional CRE within the 5'-flanking region of the gene. PMID- 12581878 TI - Differential regulation of tyrosine hydroxylase and estradiol receptor expression in the rainbow trout brain. AB - In numerous fish species, dopamine has been found to strongly inhibit gonadotropin release. Among the enzymes that regulate dopamine turnover, tyrosine hydroxylase (TH), the rate-limiting anabolic enzyme, could be a target for endocrine feedback regulation. Since dopamine turnover is stimulated by estradiol in rainbow trout, we have investigated the effect of estradiol on TH and estradiol receptor expression. In situ hybridization was used to quantify mRNA levels in the brain of ovariectomized female rainbow trout implanted or not with estradiol pellets. We demonstrated that preoptic TH and estradiol receptor mRNA levels are greatly decreased by gonadectomy during vitellogenesis. For TH expression, this effect was reversed in part by estradiol supplementation. We have also confirmed the existence of an inhibitory gonadal feedback on FSH secretion, mediated by estradiol. The stimulating effect of estradiol on TH expression found in this study could be a pathway involved in gonadal feedback on gonadotropin release. PMID- 12581879 TI - Triiodothyronine is a survival factor for developing oligodendrocytes. AB - Thyroid hormone plays an important role in oligodendrocyte development. The studies presented here suggest that thyroid hormone is required for oligodendrocyte survival during development. Oligodendrocyte precursor cells, astrocytes and microglia were cultured in a defined media. Oligodendrocyte precursor cell differentiation was induced by growth factor removal. Time course studies revealed that oligodendrocytes cultured in the presence or absence of triiodothyronine (T3) develop similarly during the first 3 days of development. Oligodendrocytes cultured in the absence of T3, however, die after developmental day 3. TdT-Mediated dUDP Nick End Labeling assay and Hoechst staining indicate that T3 rescues developing oligodendrocytes from death by apoptosis. Apoptosis is likely induced by the presence of the cytokines TNFalpha and IL-1beta. However, expression of these cytokines is not altered by thyroid hormone administration. Thus, thyroid hormone has been demonstrated to effect proliferation, myelin gene expression and now the survival of developing oligodendrocytes. PMID- 12581880 TI - Sequences required for the transition from monomeric to homodimeric forms of thyroid hormone receptor alpha and v-erbA. AB - Thyroid hormone receptor alpha (TRalpha) and the oncoprotein v-erbA (a mutated form of TRalpha incapable of binding T3) bind as heterodimers with retinoid X receptor (RXR) to DNA sequences with different orientations of AGGTCA half sites. v-erbA can also form homodimers, whereas, TRalpha1 homodimerizes poorly. Therefore, in order to obtain a better understanding for the distinct homodimerization properties between TRalpha1 and v-erbA, we created chimeras between these two receptors and tested their abilities to homodimerize on direct and everted repeats (DRs, ERs). We found that the enhanced homodimerization properties of v-erbA compared to TRalpha1 map to isoleucine at position 339 in conjunction with serine at position 351 and alanine at position 358. Our data indicate that the methyl group in isoleucine at position 339 plays an important role in v-erbA homodimerization, particularly on ER 6. Functional studies with I339V+S351P+A358T, a v-erbA mutant unable to homodimerize but still able to heterodimerize with RXR on ERs and DRs, indicate that v-erbA-RXR heterodimers mediate the dominant negative activity of v-erbA on DRs. However, the repressor activity of this mutant is weaker than that of the wild type v-erbA on ERs, suggesting that v-erbA homodimers rather than v-erbA-RXR heterodimers mediate the potent dominant negative activity of v-erbA on ERs. PMID- 12581882 TI - A Galphas protein-coupled membrane receptor, distinct from the classical oestrogen receptor, transduces rapid effects of oestradiol on [Ca2+]i in female rat distal colon. AB - We examined the hypothesis whether rapid non-genomic effects of oestradiol (E2) on [Ca(2+)](i) are mediated via a membrane-located oestrogen receptor (ER) and further elucidated the signalling pathways involved in rapid non-genomic effects of E2 on [Ca(2+)](i) in distal colonic crypts. Basal [Ca(2+)](i) was significantly increased, within minutes, in response to physiological concentrations of E2. Oestradiol linked to bovine serum albumin (E2-BSA), which renders the E2 membrane impermeable, rapidly increased [Ca(2+)](i) suggesting mediation by a membrane surface receptor. A classical ER is not involved however, as no inhibition of either the E2 or E2-BSA [Ca(2+)](i) response was seen in the presence of the classical ER antagonist ICI 182,780. Treatment with the Galphas inhibitor cholera toxin abolished both E2 and E2-BSA induced Ca(2+) increases. In contrast, treatment with pertussis toxin, an inhibitor of Galphai and Galphao, had no inhibitory effect. Following subsequent additions of E2 and E2-BSA, no further increases in [Ca(2+)](i) were observed, indicating receptor desensitisation. The E2-induced increase in [Ca(2+)](i) was completely abolished by the PKCdelta-specific inhibitor rottlerin, whereas Go6976, an inhibitor of Ca(2+)-sensitive PKC isoforms, was without inhibitory effect. The phospholipase A2 antagonist, quinacrine, and the COX1 inhibitor, indomethacin, abolished the E2 induced increase in [Ca(2+)](i). MAP kinase activation is not involved in rapid stimulatory effects of E2 on [Ca(2+)](i) as the specific inhibitor PD98059 did not inhibit the E2 response. These results demonstrate that rapid E2-induced stimulation of [Ca(2+)](i), in femal rat distal colonic crypts, occurs via a CTx sensitive Galphas-coupled membrane receptor distinct from the classical ER. PKCdelta and fatty acids are involved in the E2 signalling pathway. In contrast, PKCalpha and MAP kinase are not required. PMID- 12581881 TI - Inositol phosphate stimulation by LH requires the entire alpha Asn56 oligosaccharide. AB - Lentil lectin-bound, fucose-enriched hTSH was reported to stimulate both cAMP and inositol phosphate (IP) intracellular signalling pathways, whereas fucose depleted hTSH stimulated only the cAMP pathway. Gonadotropins activate the cAMP pathway and in several studies higher concentrations activate the IP pathway. Since only the 10% of alpha subunit Asn(56) oligosaccharides (Asn(52) in humans) are fucosylated, the higher glycoprotein hormone concentrations required for IP pathway activation might be related to the abundance of competent hormone isoforms. Lentil lectin-fractionated equine (e)LHalpha and eFSHalpha preparations were combined with a truncated, des(121-149)eLHbeta preparation. All four hybrid hormone preparations induced IP accumulation in porcine theca cells, suggesting that activation of the IP pathway was not dependent on fucosylation at alpha subunit Asn(56). However, the presence of Asn(56) carbohydrate was necessary for increased IP accumulation. Intact, rather than Asn(56)-deglycosylated eLH preparations provoked a biphasic steroidogenic response by rat testis Leydig cells, suggesting that Galpha(i) stimulation was also sensitive to loss of Asn(56) carbohydrate. While rat granulosa cells responded to human FSH preparations in a biphasic manner, a classical sigmoidal response was obtained to eFSH and Asn(56)-deglycosylated eFSH, suggesting that the equine preparations did not activate Galpha(i). Purified oLHalpha Asn(56) oligosaccharides inhibited FSH stimulated steroidogenesis in rat granulosa cell cultures indicating a direct role for carbohydrate in FSH action. The same carbohydrate preparation inhibited hCG-stimulated fluorescence energy transfer suggesting oligosaccharide involvement in activated LH receptor self-association. PMID- 12581883 TI - Endocrine disrupting chemicals: interference of thyroid hormone binding to transthyretins and to thyroid hormone receptors. AB - We examined the effects of industrial, medical and agricultural chemicals on 3,5,3'-L-[125I]triiodothyronine ([125I]T(3)) binding to transthyretins (TTRs) and thyroid hormone receptors (TRs). Among the chemicals investigated diethylstilbestrol (DES) was the most powerful inhibitor of [125I]T(3) binding to chicken and bullfrog TTR (cTTR and bTTR). Inhibition of [125I]T(3) binding was more apparent in cTTR than bTTR. Scatchard analysis revealed DES, pentachlorophenol and ioxynil as competitive inhibitors of [125I]T(3) binding to cTTR and bTTR. However, cTTR's affinity for the three chemicals was higher than its affinity for T(3). A miticide dicofol (10(-10)-10(-7) M) activated [125I]T(3) binding to bTTR up to 170%. However, at 4x10(-5) M it inhibited [125I]T(3) binding by 83%. Dicofol's biphasic effect upon [125I]T(3) binding was not detected in TTRs from other species. DES and pentachlorophenol, in the presence of plasma, increased cellular uptake of [125I]T(3) in vitro, by displacing [125I]T(3) from its plasma binding sites. These chemicals did not, however, affect the association of cTTR with chicken retinol-binding protein. All chemicals investigated had little or no influence on [125I]T(3) binding to chicken TR (cTR) and bullfrog TR (bTR). Several endocrine disrupting chemicals that were tested interfered with T(3) binding to TTR rather than to TR. Binding of the endocrine disrupting chemicals to TTR may weaken their intrinsic effects on target cells by depressing their free concentrations in plasma. However, this may affect TH homeostasis in vivo by altering the free concentrations of plasma THs. PMID- 12581884 TI - Molecular cloning and expression of equine calcitonin, calcitonin gene-related peptide-I, and calcitonin gene-related peptide-II. AB - In this study, we describe the cloning and tissue expression of equine calcitonin (CT), calcitonin-gene related peptide (CGRP)-I, and CGRP-II cDNA. We also describe a novel divergent form of CGRP (CGRP-I). Equine CT has greatest homology (>85%) to human, rat and mouse subgroups of calcitonins. Equine CGRP-I has low homology (<59%) to CGRPs of other species. The signal and N-terminal peptides for equine CT and CGRP-I were identical, indicating that these peptides are encoded by a gene equivalent to the human CALC-I gene. Equine CGRP-II has >80% homology to chicken, human, rat, ovine, swine, and bovine CGRPs. The homology between equine CGRP-I and CGRP-II is low (56%). The high homology of equine CGRP-II and the low homology of equine CGRP-I to CGRP in other species were unexpected findings. Northern blot analysis revealed that CT mRNA expression was restricted to the thyroid gland; however, RT-PCR revealed that CT mRNA expression was also present in the pituitary gland and in the liver. CGRP-I and CGRP-II mRNA expression was present in several regions of the nervous system and other tissues of neuroectodermal origin. An unexpected finding was CGRP-I expression in the kidney by both Northern analysis and by RT-PCR. Based on these results, CT gene expression in the horse was not restricted to the thyroid gland, and CT may be important in regulating pituitary cell function. CGRPs are widely expressed in tissues of the central and peripheral nervous system. Information from this study will be valuable to study the role of CT, CGRP-I, and CGRP-II in equine health and disease. PMID- 12581885 TI - A second pathway for modulating glucocorticoid receptor transactivation properties. AB - We recently reported that three factors (a cis-acting element and changing concentrations of receptor or coactivator TIF2) act at a common rate-limiting step to modulate the position of the dose-response curve and the partial agonist activity of glucocorticoid receptors (GRs). The ability of saturating levels of GR, and added inhibitors, to prevent the actions of the three modulators (cis acting element, GR, and TIF2) but not the currently investigated C-terminal fragment of E1A-13S (E1A-133C) indicates that E1A-133C alters GR properties via a second pathway that is downstream of the common step for the original three modulators. hSur2 binds to E1A-133C. We find that hSur2 modulates GR transactivation properties, thus suggesting that the effects of E1A-133C are due to the recruitment of hSur2. hSur2 also modifies GR activities in the presence of saturating GR concentrations, which is consistent with hSur2 acting downstream of the common step for the original three modulators. The H160Y mutation, which eliminates hSur2 binding to E1A, blocks most of the activity of E1A-133C. This suggests that the modulatory activity of E1A-133C is largely due to the binding of hSur2, which is a component of the Mediator complex. Collectively, these data support the existence of a new pathway for modulating GR transactivation processes, thereby increasing the number of cellular mechanisms that permit differential control of gene expression by endogenous levels of glucocorticoid hormones. PMID- 12581886 TI - Nongenomic estrogen action regulates tyrosine phosphatase activity and tuberin stability. AB - Estrogen action and tuberin function has been suggested to play a crucial role in the proliferation of lung smooth muscle-like cells and/or myofibroblasts in pulmonary lymphangioleiomyomatosis (LAM). Tuberin is a tumor suppressor phosphoprotein, which also regulates fluid phase endocytosis. Its activity, turnover and complex association with hamartin depends on its phosphorylation status. We have recently reported that nongenomic estrogen action regulates the phosphorylation status of several cytoplasmic proteins. Herein, we demonstrate that estrogen increases tyrosine phosphatase activity, which can be abrogated by antiestrogen ICI 182780 and tyrosine phosphatase inhibitor bpV(phen), but not by the protein synthesis inhibitor cyclohexamide. Furthermore, we show that estrogen transiently enhances the turnover of tuberin, which follows an inverse pattern to that observed for tyrosine phosphatase and endocytosis activity. We showed that tuberin phosphorylation protects it from degradation and induces its accumulation in female human lung fibroblasts and myofibroblasts. Our results suggest that nongenomic estrogen action induces tyrosine phosphatase activity that regulates stability of tyrosine phosphorylated proteins, including tuberin, which may play a crucial role in cellular specific functions such as endocytosis. PMID- 12581887 TI - Mechanisms of glucocorticoid-induced Leydig cell apoptosis. AB - The high levels of corticosterone (CORT) that are typically achieved during stress induce apoptotic death of Leydig cells. The intracellular mechanisms by which CORT acts on Leydig cells to induce apoptosis are unknown, and the present study tested for mediation by Fas ligand (FasL), a member of the tumor necrosis factor ligand family, in association with caspase activation. In addition, another apoptotic pathway involving in the participation of mitochondria was studied by evaluation of mitochondrial membrane potential (DeltaPsi) loss and generation of reactive oxygen species (ROS), which are early apoptotic events in many cell types. Rat Leydig cells were isolated from adrenalectomized rats on day 90 postpartum at 3, 6, 12, 24 and 48 h after the start of CORT administration (at a dose of 5 mg total/100 g body weight per day intraperitoneally in two daily injections starting 3 days after surgery). Both FasL and Fas receptor protein levels, analyzed by Western blot and fluorescent immunohistochemistry, increased at 6 h after the start of CORT administration, peaking at 24 h and declining thereafter. Leydig cell caspase-3 activity was analyzed in vitro. Low molecular weight DNA fragments that are characteristic of apoptosis were evident in Leydig cells by 12 h of exposure to 100 nM CORT in vitro, and the abundance of the fragments was more pronounced at 24 h. In the presence of a specific caspase inhibitor, Ac-DEVD-CHO, Leydig cell apoptosis was suppressed, corroborating the hypothesis that caspase-3 is involved in CORT-mediated cell death. Western blotting analysis revealed that procaspase-3 was present only at low levels in untreated control Leydig cells, and increased by 6 h of CORT administration. By 12 h, however, procaspase-3 was significantly reduced, and the cleaved, active caspase-3 forms appeared and increased through 24 h. These results indicated that FasL/Fas and caspase were implicated in CORT-mediated Leydig cell apoptosis. Decreased DeltaPsi and increased ROS generation were also measurable in Leydig cells for up to 2 days following CORT administration in vitro. These data indicate that activation of the Fas system, cleavage of procaspase-3, loss of DeltaPsi and increased ROS generation are all implicated in the process of CORT induced Leydig cell death. PMID- 12581888 TI - PTH-related protein modulates PC-3 prostate cancer cell adhesion and integrin subunit profile. AB - Parathyroid hormone-related protein (PTHrP), which has been localized in prostate cancer tissue and cell lines, plays a role in the development of bone metastases, a frequent complication in prostate cancer patients. Tumor cell adhesion to extracellular matrix (ECM) components is mediated via integrin subunits, and plays a major role in the invasion and metastasis of tumor cells. The present experiments examined the ability of PTHrP to influence adhesion of the human prostate cancer cell line PC-3 to several ECM proteins found in normal tissues. Clonal PC-3 cells induced to overexpress PTHrP by stable transfection with PTHrP complementary DNA showed significantly higher adhesion to collagen type 1, fibronectin, and laminin than control (empty vector-transfected) cells. PTHrP overexpressing cells also exhibited higher expression of the alpha1, alpha5, alpha6, and beta4 integrin subunits. These results suggest that PTHrP may play a role in prostate tumor invasion and metastasis by influencing cell adhesion to the ECM via upregulation of specific integrin subunits. PMID- 12581889 TI - Overexpression of adrenomedullin gene markedly inhibits proliferation of PC3 prostate cancer cells in vitro and in vivo. AB - The expression of the gene encoding adrenomedullin (AM), a multifunctional peptide hormone, in the prostate is localized to the epithelial cells. Prostate cancer cells are derived from prostatic epithelial cells. To elucidate the potential role of the AM gene in prostate cancer progression, we have stably transfected the PC3 human prostate cancer cell line with an AM gene expression vector. The AM-transfected PC3 sublines were studied along with parental and empty vector transfected PC3 cells as controls. The average level of AM in the conditioned media of AM-transfected cells was 0.959+/-0.113 nM, a physiologically relevant concentration. The ectopic expression of AM gene inhibited the proliferation of PC3 cells in culture dishes. In addition, anchorage-independent growth of the transfected sublines was virtually abolished in soft agar assays. Flow cytometry studies showed that overexpression of AM gene caused a very significant G(1)/G(0) cell cycle arrest. In vivo experiments demonstrated that AM gene expression markedly inhibited the growth of xenograft tumors in nude mice. Our in vivo and in vitro studies suggest that AM could strongly suppress the malignancy of prostate cancer cells, via autocrine and/or paracrine mechanisms. PMID- 12581890 TI - Human corticotropin-releasing factor type 1 receptor analysis with white blood cells mRNAs and DNA. AB - The human corticotropin-releasing factor receptor type 1 (hCRF-R1) functional transcript is mainly expressed in the anterior pituitary corticotrophs, a tissue usually not available for clinical investigation. Splice variants translated into defective isoforms of the receptor have been described in few peripheral tissues. The aim of this work was to determine whether peripheral white blood cells from healthy individuals, an accessible tissue for clinical investigation, were suitable for the analysis of the hCRF-R1 transcript and gene. We report that: (i) specific amplification of the hCRF-R1 transcript from peripheral white blood cells mRNAs is feasible; (ii) this transcript is similar to the functional transcript; (iii) the draft sequence of chromosome 17 and unrelated sequences allow direct sequencing of all 14 exons of the gene, adjacent splice sites and related branch points. In conclusion, these approaches would be suitable for studies in patients having isolated secondary glucocorticoids deficiency to implicate the hCRH-R1 in the etiology of the disease. PMID- 12581891 TI - Acute lymphoblastic leukemia characterized by t(8;14)(q11.2;q32). AB - The t(8;14)(q11.2;q32) is emerging as an uncommon, though recurrent cytogenetic finding. As of yet, too few cases of acute lymphoblastic leukemia (ALL) characterized by this translocation have been studied to determine its prognostic significance with confidence. We therefore report three new patients (two male children and one adult female) and present their hematologic, immunophenotypic, and clinical data. The clinical and laboratory characteristics of 26 other patients with t(8;14)(q11.2;q32) are summarized. The total number of patients now reported in the literature is 29 with a mean age of 14 years. Early relapse, that is, relapse within 6 months, does not appear to be a common feature of this group. The gender distribution is 19 males: 9 females (gender not reported in one case). Twenty-three t(8;14) patients show a pre-B immunophenotype and 24 of 24, on whom information is available, achieved complete remission after induction chemotherapy for B-ALL. Approximately one third of patients with t(8;14) have Down syndrome, 19 of 27 have additional acquired cytogenetic abnormalities, 5 of these have the t(9;22), and 4 show duplication of the abnormal chromosome 14, which is derived from the t(8;14). Hemoglobin and platelet counts are low at presentation in 10 of 10 and 8 of 9 patients, respectively, and the average white blood count is 38.9 x 10(9)/L. Of the 7 patients for whom IgH status has been determined, all show rearrangement of the IgH locus. Two of the present three patients are included in this group; their IgH rearrangement was demonstrated by fluorescence in situ hybridization with IgH break-apart probes. PMID- 12581892 TI - Pathogenetic and biologic significance of TP14ARF alterations in nonsmall cell lung carcinoma. AB - The INK4a/ARF locus on human chromosome band 9p21 carries two tumor suppressor genes, TP14ARF and TP16INK4a, and both are frequently inactivated in nonsmall cell lung carcinoma (NSCLC. TP14ARF and TP16INK4a play important roles in the TP53 and RB tumor suppressor pathways, respectively. To elucidate the genetic and epigenetic status of the TP14ARF and TP16INK4a genes in NSCLC, we comprehensively analyzed mutations, homozygous deletions, methylations in the CpG regions, and expression of the TP14ARF and TP16INK4a genes in 31 NSCLC cell lines. TP16INK4a (84%) was inactivated more frequently than TP14ARF (55%). Moreover, p16INK4a was inactivated in all 17 cell lines with TP14ARF inactivation. Three cell lines with base substitutions in exon 2 resulted in missense mutations of TP16INK4a but silent mutations of TP14ARF. There was a case of mutation in exon 1alpha unique to TP16INK4a, but not a mutation in exon 1beta unique to TP14ARF. The TP16INK4a gene was methylated in 6 cell lines, but the TP14ARF gene was not methylated in any cell line. Unlike a mutually exclusive relationship for inactivation between TP16INK4a and RB, TP14ARF and TP53 did not show such a relationship (P = 0.61, Fisher exact test). Thus, the present results indicate the TP16INK4a gene to be the primary target of INK4a/ARF locus alterations. Transient TP14ARF expression induced G1 arrest in the cells with wild-type TP53, but not in the cells with mutated TP53. Thus, the pathogenetic and biologic significance of TP14ARF inactivation is different between NSCLC cells with wild-type TP53 and those with mutated TP53. PMID- 12581893 TI - Cytogenetic study of non-Hodgkin lymphoma from South India. histologic and geographic correlations. AB - Cytogenetic analysis of fine needle aspiration cultures was performed on 189 patients with non-Hodgkin lymphoma from South India. Successful karyotyping was possible in 97 patients (51.3%). Burkitt lymphoma constituted 56% of the cases studied followed by diffuse type 20%, follicular 8.8%, lymphoblastic 6.6%, and unclassified 6.6%. Characteristic chromosomal translocations were t(8;14)(q24;q32) [32.2%], t(8;22)(q24;q11) [10%], t(2;8)(p12;q24) [2.2%], t(14;18)(q34;q21) [3.3%], and t(11;14)(q23;q32) [2.2%]. Notable geographical variation of some structural abnormalities was the finding in the present study such as, lower frequency of t(14;18) in follicular lymphomas and higher frequency of t(8;14) in Burkitt lymphomas when compared with the Western studies. PMID- 12581895 TI - Loss of heterozygosity in follicular and papillary thyroid carcinomas. AB - In this study we aimed at investigating the incidence and the role of 3p deletions, particularly at the 3p25 approximately pter region, in follicle cell derived thyroid neoplasms, by using loss of heterozygosity (LOH) analysis. We analyzed 12 follicular adenomas (FA), 13 follicular thyroid carcinomas (FTC), and 15 papillary thyroid carcinomas (PTC) with 11 microsatellite markers for chromosome 3. One additional marker on 3q25.2 was also investigated for assessment of deletion extent on 3q. Microsatellite instability was detected at one locus in 1 of 15 PTC (7%) and at four loci in 1 of 13 FTC (8%). Loss of heterozygosity was found in 8 of 12 cases of FTC (67%), in 6 of 15 cases of PTC (40%), and in 2 of 12 FA (17%). We identified three minimal common deleted regions (CDR) involving significant sites of LOH: two in FTC (a new terminal region, of approximately 8 cM distal to D3S1620 at 3p25.3 approximately pter and the D3S1573-D3S1595 region at 3p21.2 approximately p12) and one in PTC (D3S1304 D3S1263 region at 3p25.3 approximately p24.2). The newly identified 3p25.3 approximately pter CDR seems to be specific for FTC. Our results suggest the existence of at least three distinct regions on 3p that might harbor tumor suppressor genes involved in the carcinogenesis processes of FTC and PTC. PMID- 12581894 TI - Establishment of a new human cell line (EN) with TP53 mutation derived from endometrial carcinoma. AB - We present a new cell line, EN, established from an invasive endometrioid adenocarcinoma of the uterine corpus in 50-year-old patient. The cells show rapid growth in culture with a doubling time of 24.4 hours and high migration activity. Monolayer-cultured cells were polygonal in shape and showed a tendency to pile up without contact inhibition. Subcutaneous transplantation of the EN cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma, whereas no metastasis was observed. Cultured EN cells produced tissue polypeptide antigen. Genetic and molecular analyses revealed high telomerase activity and estrogen receptor beta but not alpha expression. Using the polymerase chain reaction-single strand conformation polymorphism technique, we have screened EN cells for TP53 mutation in exons 5-8. A mobility shift was observed in this cell line in exon 8. A nucleotide insertion (CGT-->CAGT) was detected at codon 273, which resulted in a creation of a stop codon at codon 308. This cell line thus appears to represent the development of a more malignant clone with divergent receptor function and growth behavior, and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium. PMID- 12581896 TI - Combined 24-color karyotyping and comparative genomic hybridization analysis indicates predominant rearrangements of early replicating chromosome regions in neuroblastoma. AB - Neuroblastoma is characterized by several distinct genetic alterations including MYCN amplification, chromosome 1p deletion and gain of chromosome 17. Although these alterations are thought to play a crucial role in oncogenesis, to date little is known about their underlying mechanisms. In order to more precisely document these genetic alterations, we have performed a combined study of 27 neuroblastoma cell lines using 24-color karyotyping (24-CK) and comparative genomic hybridization (CGH). 24-CK detected balanced translocations in 13 cases with recurrent involvement of chromosome 8. More importantly, 144 nonreciprocal translocations were observed in the 27 cell lines, with chromosome 1 as the most frequent recipient and chromosome 17 the most frequent donor. Each cell line exhibited at least one unbalanced translocation involving 17q, with 14 cell lines demonstrating more than one such translocation. Other recurrent alterations were amplification of the 2p24 chromosome region, which encodes the MYCN oncogene, losses of 1p, 3p and 11q, and gains of 1q and 7. In most cases, CGH profiles were directly linked to the presence of unbalanced translocations with gain of the donor fragment and loss of the replaced region on the recipient chromosome. Strikingly, over 60% of the chromosome breakpoints mapped to early replicating chromosome bands, which represent around 13% of the genome. Altogether these data suggest that neuroblastoma is characterized by rearrangements that predominantly involve chromosome fragments replicating early in the S-phase. PMID- 12581897 TI - Molecular cytogenetic characterization of four commonly used cell lines derived from Hodgkin lymphoma. AB - Hodgkin lymphoma (HL) is a malignant lymphoma composed of a minority of neoplastic cells-the Hodgkin and Reed-Sternberg (HRS) cells-and a majority of nonneoplastic inflammatory cells. The low proportion of tumor cells makes genetic studies of primary neoplasia difficult. Therefore, established HL-derived cell lines are commonly used as model systems. Here we have characterized the chromosomal composition of four such cell lines: L-540, DEV, HDLM-2, and CO. Using spectral karyotyping (SKY), reversed DAPI banding, and comparative genomic hybridization (CGH), the karyotypes were characterized and previously unidentified marker chromosomes were resolved. The karyotype for CO was incompatible with the original description but showed striking similarities with the T-ALL-derived cell line CCRF-CEM, suggesting that CO had been cross contaminated and overgrown prior to arrival at our laboratory. Multiple numerical and structural abnormalities were identified in DEV and HDLM-2, as well as in L 540. Refined composed karyotypes are suggested for the cell lines studied, to be used as references for further studies of lymphoma. PMID- 12581898 TI - Fluorescence in situ hybridization for detecting TP16 MTS1/CDK41 gene deletions in squamous cell carcinoma of the head and neck. AB - We have previously shown TP16 MTS1/CDK41 gene deletion in more than 50% of a cohort of squamous cell carcinoma of the head and neck (SCCHN) patients using polymerase chain reaction (PCR). We have performed fluorescence in situ hybridization (FISH) on paraffin-embedded SCCHN specimens from the same cohort to identify the deletion of TP16 MTS1/CDK41CDK41gene. Twenty normal and 19 SCCHN specimens were studied. An alpha-satellite DNA probe specific for chromosome 9 and a cosmid probe for the TP16 MTS1/CDK41CDK41gene were used. Of the 19 tumors examined by FISH, 6 had homozygous deletions, 7 were hemizygously deleted, and the remaining 6 showed no evidence of deletion of the TP16 MTS1/CDK41 gene. None of the normal specimens showed TP16 gene deletion. Data obtained from FISH highly correlated with the PCR results for the identification of TP16 MTS1/CDK41 gene deletions. Patients with deletion of the TP16 MTS1/CDK41 gene show a greater tendency toward the development of recurrence and metastasis. PMID- 12581899 TI - A novel tumorigenic human prostate epithelial cell line (M2205): molecular cytogenetic characterization demonstrates C-MYC amplification and jumping translocations. AB - The paucity of cell lines from early-stage prostate cancer tumors has hindered the recognition of genetic and cellular changes that are associated with the acquisition of tumorigenesis. We describe the chromosomal complement of a novel tumorigenic prostate epithelial cell subline, called M2205, that acquired only three new, consistent chromosomal changes (from those present in the SV40T antigen immortalized parental cell line, P69SV40TAg) when it attained tumor forming potential. The consistent changes, which were fully characterized using GTG-banding, CBG-banding, silver staining, fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH), and spectral karyotyping (SKY), involved segmental jumping translocations and resulted in gains in the copy number of genes located on the distal long arm of chromosome 8 (8q22 to 8q24.3), including c-myc. Furthermore, the jumping translocations also resulted in ribosomal genes being present in multiple, tandem copies next to the chromatin from 8q. Given the relatively small number of cytogenetic changes present, this subline provides a means for better understanding the cellular changes associated with the acquired chromosomal imbalances. Further studies of this subline could also provide insight as to the mechanism or mechanisms leading to the formation of jumping translocations, as well as potential position effects resulting from the relocation of ribosomal genes next to other cellular genes or oncogenes. PMID- 12581900 TI - Novel mutations of the APC gene in familial adenomatous polyposis in Greek patients. AB - Familial adenomatous polyposis (FAP), a premalignant clinical entity inherited as an autosomal dominant trait, is characterized by the development thousands of adenomatous polyps of the colorectum during the 2nd and 3rd decade of life. Approximately 80% of patients with FAP harbor truncating germline mutations in the adenomatous polyposis coli (APC) tumor suppressor gene. We tested 24 members of six Greek families. All patients had the FAP phenotype, and one patient had an extracolonic tumor (medulloblastoma). Our method for testing was the polymerase chain reaction (PCR) amplification from genomic DNA extracted from whole blood, followed by automated DNA sequencing. Two novel truncating mutations (2601delGA and R923X) and three already-known mutations (R876X, Q1045X, and D1822V) were found. Other polymorphisms were also found. We identified the inactivating APC mutation in 12 of 13 of our FAP patients. Our results suggest that PCR sequencing is a reliable method for screening the APC gene for germline mutations. PMID- 12581901 TI - Translocation (11;19)(q23;p13.3) associated with a novel t(5;16) (q13;q22) in a patient with acute myelocytic leukemia. AB - A novel association of t(11;19)(q23;p13) and t(5;16)(q13;q22) was detected by G banding and spectral karyotyping studies in an 18-year-old patient. While balanced t(11;19) has been often described in acute myelocytic leukemia (AML) French-American-British Cooperative Group subtypes M4 and M5, this patient was diagnosed with the variant AML-M4 with eosinophilia (AML-M4Eo), which is associated with abnormalities in 16q22 and has good prognosis. However, the patient relapsed after allogeneic transplant and died within 2 years of diagnosis, which suggests that the association of these two translocations correlates with a poor prognosis. This report expands the molecular basis of the variability in clinical outcomes and adds the novel t(5;16)(q13;q22) to the spectrum of chromosome 16q22 abnormalities in AML. PMID- 12581902 TI - The use of clustering software for the classification of comparative genomic hybridization data. an analysis of 109 malignant fibrous histiocytomas. AB - Malignant fibrous histiocytoma (MFH) is considered the most frequent soft-tissue sarcoma of late adult life. Nevertheless, the validity of this entity has been recurrently questioned by pathologists. Preliminary analyses by comparative genomic hybridization (CGH) of series of MFH have suggested that this tumor group is heterogeneous at the genomic level, and that at least two main genetic subgroups exist. We report an analysis by CGH of a large series of 109 MFH and on the use of clustering software for an objective classification of these tumors. We confirm our preliminary CGH results and demonstrate that two main clusters of tumors are present in the series analyzed. PMID- 12581903 TI - Pentasomy 8q in therapy-related myelodysplastic syndrome due to cyclophosphamide therapy for fibrosing alveolitis. AB - Trisomy 8/8q is a common cytogenetic event in myelocytic malignancies, ranging from myelodysplastic syndrome (MDS) to acute myelocytic leukemia (AML) to blastic transformation of chronic myelocytic leukemia. Isochromosome 8q results in the same gene dosage effect. Duplication of i(8q), resulting in pentasomy 8q, has been reported only in two cases of AML. A patient with fibrosing alveolitis on prolonged cyclophosphamide treatment developed therapy-related MDS. Karyotyping, FISH, and CGH analysis showed a duplicated i(8q) among other complex abnormalities. The clinical features of 11 cases of myelocytic leukemia with pentasomy and hexasomy 8/8q were summarized. Compared with trisomy and tetrasomy 8, significant features included reduced median survival (90 days), treatment refractoriness (even with transplantation), monocytic differentiation, trilineage dysplasia, and radiation or toxin exposure. Increasing copy numbers of chromosome 8/8q may therefore be a marker of advanced leukemic evolution, exposure to toxins, underlying myelodysplasia, and an overall poor prognosis. PMID- 12581904 TI - Waldenstrom macroglobulinemia with a novel der(8;17)(q10;q10). AB - We report the occurrence of an unbalanced whole-arm translocation of der(8;17)(q10;q10) in an 80-year-old female patient with Waldenstrom macroglobulinemia. To our knowledge, der(8;17)(q10;q10) has not been described in Waldenstrom macroglobulinemia. Although this cytogenetic abnormality has been reported in a number of solid tumors, a literature review suggests also a possible association with B-cell chronic lymphoproliferative disorders. PMID- 12581905 TI - Burkitt lymphoma following splenic marginal zone lymphoma. evidence for two independent B-cell clones. AB - We report the progression of splenic marginal zone lymphoma (SMZL) with circulating villous lymphocytes to Burkitt lymphoma with the presence of a t(8;14)(q24;q32) followed by a highly aggressive course. While the initial indolent lymphoma had an IgM lambda immunophenotype the Burkitt lymphoma was IgM kappa-positive. Immunoglobulin heavy chain gene (IGH) sequence analysis showed no identity between the two clones. We conclude that Burkitt lymphoma can occur in patients with SMZL, although not necessarily of identical clonal origin. PMID- 12581906 TI - Simultaneous detection of deletions of 9q and 22q in a subgroup of chronic myelocytic leukemia Philadelphia-positive patients by a novel probe. PMID- 12581930 TI - Exercise self-efficacy in older adults: social, affective, and behavioral influences. AB - A 6-month randomized controlled trial examined the effect of walking and stretching/toning activity on changes in self-efficacy to overcome barriers and engage in incremental periods of activity in older, formerly sedentary adults (N = 174, M age = 65.5 years). Additionally, we were interested in the extent to which social, affective, and behavioral influences contributed to self-efficacy at the end of the 6-month program. Multiple sample latent growth curve analyses revealed a nonsignificant curvilinear growth pattern for barriers efficacy with increases in efficacy occurring from baseline to 2 months and then declining at 4 and 6 months. In the case of efficacy related to continued activity participation, there was a significant growth pattern demonstrating declines in efficacy over the 4 time points. Structural modeling analyses revealed significant direct effects of physical activity, affect experienced during activity, and exercise social support on both types of self-efficacy. These relationships were not significantly different between modes of activity. The findings are discussed in terms of the need to target sources of efficacy information prior to program end and the implications that such an approach might have for long-term maintenance of physical activity in older adults. PMID- 12581931 TI - How do female and male cardiac patients and their spouses share responsibilities after discharge from the hospital? AB - This study explored sex differences in household and employment responsibilities among cardiac patients (N = 63; 46 men) and spouses during the 5 months following discharge from the hospital. Results showed that both patients and partners maintained traditional sex-typed activities. As patients or spouses, women tended to assume greater responsibility for domestic tasks such as laundry, cleaning, and cooking than their husbands. Men as patients or spouses tended to assume greater responsibility for household repair and maintenance tasks. Husbands also worked more for pay outside the home than did wives, except in couples where the male patient was high risk. Correlations for male patients indicated that reports of more cardiac symptoms were associated with assuming fewer responsibilities. In contrast, among women, the correlations between symptoms and activities were more complex and suggested that female patients might not be heeding signs of overexertion. PMID- 12581932 TI - The Concerns About Recurrence Scale (CARS): a systematic measure of women's fears about the possibility of breast cancer recurrence. AB - The Concerns About Recurrence Scale (CARS) systematically assesses the extent and nature of women's fears about the possibility of breast cancer recurrence. In this study with 169 breast cancer survivors, scores on the CARS indicate moderate levels of fear about recurrence overall and demonstrate a range in levels of fear. Findings further suggest that women's fears of recurrence center around the possibility of death, future treatment, and threats to health more than issues related to roles, femininity, sexuality, or body image. Younger women and women who have had chemotherapy demonstrate greater fears. There is little evidence that cancer stage, time since diagnosis, or type of surgery relate to fears. The CARS was found to be internally consistent, and there is preliminary evidence of its validity, although future research is needed. The measure will likely be a useful tool for researchers and clinicians seeking to understand women's fears about the possibility of breast cancer recurrence. PMID- 12581933 TI - Enhanced adrenocortical responses to stress in hypertension-prone men and women. AB - Hypertension risk may be associated with increased adrenocortical activity, but the extent to which this enhanced activation differs between men and women at rest and in response to psychological stress is not known. This study examined gender differences in adrenocortical responses to an extended public-speaking stressor in persons at high (resting systolic blood pressure > median; n = 21) or low risk (negative parental history and < or = median systolic blood pressure; n = 26). Salivary cortisol levels were assessed at rest and in response to public speaking in a repeated measure design on two test sessions held on separate days. Heart rate, systolic blood pressure, and diastolic blood pressure were obtained at 3-min intervals before, during, and after the task. High-risk participants showed greater cortisol, blood pressures, and heart rate responses to the public speaking stressor than the low-risk group (ps < .01). Men showed greater cortisol concentrations than women (p < .05), independent of hypertension risk status. Cardiovascular measures during the acute stressor predicted subsequent cortisol production, but only in the high-risk group. Results suggest that hypertension risk is associated with enhanced physiological reactivity across sympathetic and adrenocortical systems, supporting the possibility that this exaggerated reactivity may represent a marker of risk in hypertension-prone men and women. PMID- 12581934 TI - Defensiveness and 3-year blood pressure levels among young adults: the mediating effect of stress-reactivity. AB - Stress-reactivity has long been proposed to act as a possible mechanism linking psychological factors and increases in blood pressure (BP). This study used results from an intensive cardiovascular investigation to examine the mediating role of reactivity in the relationship between defensiveness and 3-year ambulatory BP levels. A total of 125 male and female participants (M = 29.1 years) completed a protocol including laboratory reactivity testing, 8- to 12-hr ambulatory BP monitoring, and standardized response style instruments. Further, participants returned 3 years later for an identical second test protocol. Results indicated that high- and low-defensiveness participants did not differ on Year 1 BP, but the high-defensiveness group showed higher BP during the reactivity tasks and on Year 3 ambulatory measures. Statistical mediation testing supported reactivity levels as an intervening mechanism between defensiveness and 3-year BP levels. PMID- 12581935 TI - Interleukin-1beta elevation during the postpartum period. AB - During the postpartum period, women frequently report increased fatigue, which, if severe, may interfere with maternal-child bonding, delay a new mother's return to her activities of daily living, and contribute to depression. Several studies have sought to determine psychosocial contributions to fatigue during the postpartum period, but few evaluate any physiological changes that may contribute to fatigue during this time. The following study was designed to test whether the potent, pro-inflammatory cytokine interleukin-1beta (IL-1beta), known to be a physiological mediator of fatigue in several clinical and experimental conditions, is elevated in women during the postpartum period and whether it might be related to symptoms of fatigue. Levels of fatigue and the urinary excretion of IL-1beta were measured in 26 women over 4 weeks postpartum. Correlations between fatigue and activation of the inflammatory response were investigated. Results demonstrated a significant elevation in IL-1beta during the postpartum period compared to control participants (p < .05) and a significant, although delayed, correlation between IL-1beta elevation and fatigue (p < .05). These results suggest that activation of the inflammatory response, as reflected by elevation in urinary IL-1beta, occurs in association with postpartum fatigue. Studies to explore further this association and to identify specific mechanisms of action are needed. PMID- 12581936 TI - Expectations of chemotherapy-related nausea: emotional and experiential predictors. AB - Despite literature indicating that expectations are important determinants of experiences of nonvolitional outcomes (e.g., pain, nausea), little research has explored their source. The dual process model suggests that concurrent experience of emotional distress should be a strong contributor, whereas social learning theory supports the position that expectations of nonvolitional outcomes should be based primarily on prior experience. These alternative hypotheses were tested in a sample of 80 breast cancer patients undergoing outpatient adjuvant chemotherapy, as such protocols provide a unique opportunity to study the impact of emotional distress and prior experience on patients' expectations of side effects. Bivariate analyses suggested that emotional distress contributed early in the course of treatment, but once prior experience of posttreatment nausea (PTN) was accounted for in the statistical model, distress no longer made significant contributions to patients' expectations of PTN, whereas prior experience did (p < .001). These findings suggest that experience of chemotherapy infusions is an important source of expectations for PTN, which may be representative of the source of expectations for nonvolitional outcomes in broader contexts. PMID- 12581937 TI - Perceived racism and blood pressure: a review of the literature and conceptual and methodological critique. AB - Racial disparities in health, including elevated rates of hypertension (HT) among Blacks, are widely recognized and a matter of serious concern. Researchers have hypothesized that social stress, and in particular exposure to racism, may account for some of the between-group differences in the prevalence of HT and a portion of the within-group variations in risk for HT. However, there have been surprisingly few empirical studies of the relationship between perceived racism and blood pressure (BP) or cardiovascular reactivity (CVR), a possible marker of mechanisms culminating in cardiovascular disease. This article reviews published literature investigating the relationship of perceived racism to HT-related variables, including self-reported history of HT, BP level, or CVR. Strengths and weaknesses of the existing research are discussed to permit the identification of research areas that may elucidate the biopsychosocial mechanisms potentially linking racism to HT. We hope to encourage investigators to invest in research on the health effects of racism because a sound and detailed knowledge base in this area is necessary to address racial disparities in health. PMID- 12581938 TI - Cognitive behavioral stress management effects on injury and illness among competitive athletes: a randomized clinical trial. AB - Cognitive behavioral stress management (CBSM) has previously been found to reduce fatigue, depression, and cortisol response to heavy exercise training among competitive collegiate athletes and to speed physical and psychological recovery from surgery. Our study assessed the efficacy of a CBSM program to reduce the frequency of injury and illness among collegiate athletes in a randomized, single blind, controlled clinical trial. Following assessment of baseline medical history, mood state, stress, cortisol, sleep, alcohol use, and exercise training, collegiate rowers were stratified by gender and competitive level and randomly assigned to either a control group or a CBSM group. Exercise training information and psychosocial assessments were repeated immediately following the intervention period, and health care providers who were blinded to participant assignment recorded the frequency of medical visits and the number of days injured or ill until the end of the season. Athletes randomly assigned to a CBSM group experienced significant reductions in the number of illness and injury days as compared to control group athletes. CBSM participants also had half the number of health service visits as did controls. The data suggest that a time-limited CBSM intervention designed specifically for an athlete population may be an effective prophylactic treatment to reduce the incidence of injury and illness among competitive collegiate athletes. PMID- 12581939 TI - Addressing health disparities in middle school students' nutrition and exercise. AB - Those with low income, especially women of African American and Hispanic heritage have the greatest risk of inactivity and obesity. A 4-session (Internet and video) intervention with healthy snack and gym labs was tested in 2 (gym lab in 1) urban low-middle-income middle schools to improve low fat diet and moderate and vigorous physical activity.1 The gym lab was particularly beneficial (p =.002). Fat in diet decreased with each Internet session in which students participated. Percentage of fat in food was reduced significantly p =.018 for Black, White, and Black/Native American girls in the intervention group. Interventions delivered through Internet and video may enable reduction of health disparities in students by encouraging those most at risk to consume 30% or less calories from fat and to engage in moderate and vigorous physical activity. PMID- 12581940 TI - Assessing the effectiveness of a walking program on physical function of residents living in an assisted living facility. AB - The purpose of this study was to assess the effectiveness of a resident-led walking program at an assisted living facility (ALF). Seventeen women (mean age 80, range 62-99) agreed to participate in a "walking club." Pre and post measurements included the Tinetti Performance-Oriented Assessment of Mobility Problems in Elderly Patients, the Functional Reach Test, and the Barthel Index to measure independence in activities of daily living (ADLs). Participants set their own goals for walking distance and frequency, with the assistance of the lead researcher if requested. Distances ranged from 75 ft to over 1 mile. At the end of the 9-week intervention, there was a significant increase in all of the pre test measurements. In addition, post-test assessment included individual interviews with participants about their experiences with the physical activity program. Four positive themes about the walking program emerged: (a) as a pleasurable activity, (b) as a way to manage current health problems, (c) as a way to continue life-long activity, and (d) because of perceived physical and psychological benefits from the activity. The participants planned to continue the walking program. The results of this pilot study suggest that a walking program can be instituted in an assisted living facility with minimal staff effort and significant benefit to residents. PMID- 12581941 TI - The use of music to promote sleep in older women. AB - Fifty-two women over the age of 70 participated in a study to investigate the use of an individualized music protocol to promote sleep onset and maintenance. They were recruited from the practices of physicians and nurse practitioners, and met the inclusion and exclusion criteria of the International Classification of Sleep Disorders (1990), and the Diagnostic and Statistical Manual of Mental Disorders (1994). Results indicated that the use of music decreased time to sleep onset and the number of nighttime awakenings. Consequently, it increased satisfaction with sleep. Nurses may wish to recommend the use of music at bedtime to older women with insomnia. PMID- 12581942 TI - Low-income pregnant mothers' experiences of a peer-professional social support intervention. AB - This article describes 7 low-income mothers' experiences of attending a motivational event that combined peer and professional support in a baby-shower luncheon. The event provided a group context that reinforced health-enhancing messages presented by nurses and other professional staff in home visits. Important themes that emerged in qualitative analysis include reduction of isolation and promotion of bonding; information sharing and modeling to others; and enhancement of self-esteem. A discussion of how community health nurses can use these strategies in caring for pregnant women is presented. PMID- 12581943 TI - Taking action in communities: women living with HIV/AIDS lead the way. AB - For the past 5 years, a successful collaboration of a medical center, a community based HIV/AIDS service organization, a university school of nursing, and women living with HIV/AIDS in an inner city community resulted in a series of educational programs1 for women living with HIV/AIDS, their family, friends and caregivers. These programs were intended to provide inner-city women who were living with HIV/AIDS with knowledge and new insights from the voices of their peers. Topics focused on their self-care and empowerment so that they could take control of their wellness and their health care while remaining in their community. The efforts made in launching the collaborative educational series created trusting relationships between academic, clinical, and community service agencies, professional caregivers, and the recipients of their care. The most significant contribution belonged to the women living with HIV/AIDS themselves: After the programming they turned their health and life experiences into "action" by planning a research project that will test the efficacy of an intergenerational HIV prevention program for adolescent women of color in their community in the future. PMID- 12581944 TI - Failure to maintain adherence to HAART in a cohort of French HIV-positive injecting drug users. AB - The study, carried out in the French MANIF 2000 cohort of HIV positive patients contaminated through injecting drug use, assessed the impact of patients' sociodemographic and psychological characteristics, behaviors toward drug abuse, and antiretroviral treatment characteristics on the maintenance of adherence to HAART (highly active antiretroviral therapies). A total of 96 patients (30 men and 66 women), who were initially adherent at their first visit after HAART prescription, were considered for analysis. Among these 96 patients, 22 (22.9%) experienced adherence failure defined as a self-reported, non-adherence behavior at any visit before the 18th month of treatment. Logistic regression showed that lack of a stable relationship, active drug injection, and depression were independently associated with adherence failure. Patients' counseling for facilitating maintenance of adherence to HAART over time should focus on prevention of drug use, provision of social support and consider the potential impact of difficulties with treatment on psychological well-being. PMID- 12581945 TI - Reliability and validity of the Chinese version of the Job Content Questionnaire in Taiwanese workers. AB - The reliability and validity of four selected scales--namely, job control, psychological demands, supervisor support, and coworker support--from the Chinese Version of the Job Content Questionnaire (C-JCL) were studied in 551 male and 648 female workers in Taiwan. Cronbach's alpha coefficients for job control, supervisor support, and coworker support were all above.80. Whereas it was.55 for psychological demands, indicating insufficient internal consistency for this subscale. Participants responded to questionnaire items relatively consistently over a 3-month period. Exploratory factor analyses disclosed 4 empirical factors, which corresponded closely with theoretical constructs of the JCL. As predicted by the model, lowest levels of job satisfaction were found in workers in "iso strain" category (i.e., high demands combined with low control and low social support). These findings indicated that the C-JCL is reliable and valid for assessing psychosocial work conditions among Taiwanese workers, although further improvement is needed for the psychological demands subscale. PMID- 12581946 TI - Psychometric assessment of Finnish versions of exercise-related measures of Transtheoretical Model constructs. AB - The Transtheoretical Model offers a promising approach for understanding exercise behavior. However, this model has received minimal cross-cultural attention, and measurement instruments are only available in English. This gap was addressed by assessing the psychometric properties of translated measures of each core construct from the model. Finnish (n = 168) and American (n = 169) college students completed language specific measures. Among the Finnish, American, and combined samples, the measures demonstrated acceptable reliability. Using regression analysis, 34% 18%,and 26% of the variance in the Finnish, American, and combined samples' exercise behavior, respectively, was explained by self efficacy and the behavioral processes of change, with no unique variance added by other core constructs or demographic factors. The translation of the Transtheoretical Model of behavior change scales/subscales into Finnish was successful. Within a cross-cultural sample, the translated measures demonstrated acceptable psychometric properties. PMID- 12581947 TI - Vital exhaustion in relation to lifestyle and lipid profile in healthy women. AB - "Vital exhaustion," characterized by fatigue, irritability, and demoralization, precedes new and recurrent coronary events. Biological mechanisms explaining this association are not fully understood. The objective was to investigate the relationship between vital exhaustion, lifestyle, and lipid profile. Vital exhaustion, smoking, body mass index (BMI), alcohol consumption, exercise capacity, and serum lipids were determined in 300 healthy women, aged 56.4 +/- 7.1 years. No statistically significant associations were found between vital exhaustion and lifestyle variables. Divided into quartiles, vital exhaustion was inversely related to high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A1 in a linear fashion after adjustment for age, BMI, exercise capacity, and alcohol consumption. A multivariate-adjusted vital exhaustion-score in the top quartile, as compared to one in the lowest, was associated with 12% lower HDL-C and 8% lower apolipoprotein A1 (p < .05). In conclusion, alterations in lipid metabolism may be a possible mediating mechanism between vital exhaustion and coronary heart disease. The impact of lifestyle variables was weak. PMID- 12581948 TI - Absence of an impact of emotional distress on the perception of intracardiac shock discharges. AB - The pain of transvenous cardioversion shocks in awake patients is an important clinical problem. It is unknown whether psychological factors modulate any observed variation in pain perception. Thirty-seven patients with chronic atrial fibrillation (AF; mean age 61.9 years, 29 men, 8 women) were consecutively included in the study. Pain perception of a low energy test shock (60V, 0.1J) during internal cardioversion was assessed immediately following the stimulus. Before treatment anxiety, depression, somatization were evaluated. The intracardiac shock was perceived as hypalgesic in 15 (41%) patients, as normalgesic in 10 (27%), and as hyperalgesic in 10 (32%) patients. Pain threshold was significantly lower in patients in which AF was accidentally diagnosed (p < 0.029). Age, sex, and the level of education were equally distributed over the three pain groups. The hyperalgesic pain threshold was not associated with anxiety, depression, or the patients' tendency to amplify benign bodily sensations. This study discloses a wide range of perceived pain intensity caused by a homogenous intracardiac stimulation. Anxiety and depression do not amplify perception of intracardiac shocks. Sensory gating mechanisms may play a more important role in the level of intracardiac shock perception than distressing affective factors. PMID- 12581949 TI - Biofeedback of baroreflex sensitivity in patients with mild essential hypertension. AB - An attempt was made to reduce blood pressure by increasing the baroreflex sensitivity (BRS) via biofeedback. Six patients with mild essential hypertension and 5 normotensive participants were studied during 8 biofeedback sessions. Each session consisted of 5 trials, 5 min each. The first and the last trials served as baselines of heart rate, blood pressure, respiration, and BRS. During the 3 middle trials the BRS was calculated online using the sequencing technique. The resulting value was used as a visual analogue feedback signal. Participants were asked to increase BRS. The mean BRS was 8.3 [ms/mmHg] for the hypertensive patients and 12.2 [ms/mmHg] for the normotensive participants. During biofeedback trials as well as across sessions neither the hypertensive nor the normotensive group showed a statistically significant increase of BRS, only heart rate variability increased significantly. Contrary to expectation blood pressure increased in both groups. One hypertensive subject made significant progress during the training by performing valsalva maneuvers. The data show that BRS is reduced in hypertensive subjects. The increase of the heart rate variability could be a sign of the activation of the baroreflex although the BRS itself did not increase. Despite the successful technical and organizational implementation of this biofeedback approach, it was not effective to systematically reduce blood pressure. A further development in the direction of guiding the patient to use the valsalva breathing pattern and/or a prolonged duration of the biofeedback training might be promising. PMID- 12581950 TI - Adaptive behavior in stressful situations in relation to postinfarction mortality results from prospective cohort study "Men Born in 1914" in Malmo, Sweden. AB - The serial Color Word Test, which is a semi-experimental way to differentiate adaptive behavior in stressful situations, was administered at baseline to men participating in the prospective cohort study "Men born in 1914". During follow up, from 1982-1983 until December 31, 1996, 133 men experienced a myocardial infarction. Four patterns of adaptive behavior in 2 separate dimensions, the Variability and the Regression, can be discerned during testing. These patterns were compared regarding outcome following the myocardial infarction. The Cumulative-dissociative pattern of the Regression dimension was univariately associated with mortality within 28 days (OR 5.75, CI 1.85-17.88, p = .003). Dissociative (OR 3.87, CI 1.21-12.42, p = .023) and Cumulative-dissociative (OR 5.46, CI 1.66-17.96, p = .005) patterns, of the same dimension, were independently associated with mortality within one year. Specific difficulties in adaptation to stressful situations were associated with increased risk of death following a myocardial infarction. In this male sample, these difficulties could be identified with the serial Color Word Test. PMID- 12581951 TI - Effects of prednisone on bone mineral density and biomechanical characteristics of the femora and lumbar vertebras in rats. AB - OBJECTIVE: To explore effects of prednisone on the bone mineral density (BMD) and biomechanics of the femora and lumbar vertebras in rats. METHODS: Twenty one 3 month-old male Sprague-Dawley rats weighing 226+/-12 g were randomly divided into basal control, age-matched and hormone groups. The rats in basal control group were killed at the beginning of the experiment without any treatment, and those in age-matched group were given oral normal saline (5 ml x kg(-1) x d(-1)) while the rats in hormone group received oral prednisone acetate (4.5 ml x kg(-1) x d( 1) twice a week) to establish osteoporotic models. The treatment for the latter 2 groups of rats lasted for 90 days, after which the BMD and mechanical measurements of the femurs and L5 vertebra were carried out by way of torsion, three-point bending and compression tests. The measurements were also conducted in the basal control group at the time indicated above. RESULTS: In hormone group, the total BMD of the femora and L5 vertebra was decreased by 14.64%(P<0.01), and the BMD in the right and left distal femoral segments and the vertebra decreased by 21.42% (P<0.01), 19.62% (P<0.05) and 23.48%(P<0.01), respectively, in comparison with the control group. In the meantime, the loads of three-point bending test in hormone group was decreased by 17.1%(P<0.05), whereas the rest parameters tended to decrease as compared with the control group. CONCLUSIONS: Chronic use of corticosteroid is more liable to cause bone mass loss in rat cancellous bone than in the cortical bone, and mechanical properties of the cortical and cancellous bone, especially those of the latter, will also decline, to give rise to easy bone fracture at the trabecular bone in osteoporotic conditions. PMID- 12581952 TI - [Construction of the non-resistant attenuated Salmonella typhimurium strain expressing Helicobacter pylori catalase]. AB - OBJECTIVE: To construct a non-resistant attenuated Salmonella typhimurium (S.typhimurium) strain capable of expressing Helicobacter pylori (Hp) catalase. METHODS: After PCR amplification, the gene fragment encoding Hp catalase was inserted into the expression vector pYA248 containing asd gene, and the recombinant vector was then introduced into the host S.typhimurium strain X4072 depleted of genes encoding adenylate cyclase (delta cya), cyclic adenosine monophosphate receptor protein (delta crp) and aspartate-beta-semialdehyde dehydrogenase (delta asd). Bridged enzyme-linked immunosorbent assay (ELISA) was employed to measure the antigenicity of the catalase expressed in the sonicate and culture supernatant. According to Meacock's method and with the assistance of the growth curve, the stability of the recombinant strain was evaluated. A half lethal oral dose test was conducted to evaluate the safety of recombinant strain. RESULTS: S.typhimurium X4072 (pYA248-CAT) with expected capacity was successfully constructed, and bridged ELISA demonstrated higher catalase levels in the culture supernatant than in the sonicate of the recombinant strain X4072 (pYA248-CAT). After the strain was passaged for 100 generations without selection pressure, all the randomly selected colony of the recombinant strain grew well with positive catalase antigenicity as identified by ELISA. The growth curve of the recombinant strain showed comparable growth status of the 2 strains X4072 (pYA248) and X4072 (pYA248-CAT). The survival rate of C57BL/6 mice was 100% 30 d after oral administration of 1.0x10(10) cfu X4072 (pYA248-CAT). CONCLUSION: Non-resistant S. typhimurium vaccine X4072 (pYA248- CAT) is constructed successfully, which is stable in vitro and safe as confirmed by animal experiment. This vaccine provides a new candidate for viable oral vaccine against Hp infection. PMID- 12581953 TI - [Establishment and growth regulation of rat hepatocyte colony in vitro]. AB - OBJECTIVE: To investigate the conditions essential for culturing hepatocytes colony in vitro, and to study the growth regulation mechanisms of hepatocyte colony. METHODS: Rat hepatocytes were isolated by two-step in situ preperfusion and collagenase circulatory perfusion. The effects of hepatopoietin (HPN), nicotinamide (NA) and dimethyl sulfoxide (DMSO) on DNA synthesis, mitotic activity, morphology (under inverted microscope) and utrastructure (under TEM) of the hepatocytes were investigated in chemically defined culture medium. RESULTS: The time course of DNA synthesis in cultured hepatocytes showed that 3H-TdR incorporation was dramatically enhanced by 10 mmol/L NA, presenting 2 peaks at 60 and 84 h respectively. The plateau of DNA synthesis was diminished in the presence of DMSO, but a peak occurred again at 132 h upon NA treatment. After cell culture in the presence of HPN, NA, and DMSO for 72 h, the hepatocytes presented sustained regular bipolar mitosis, with considerable mitotic activity at 168 h. The growth characteristics of hepatocyte colony, in addition to its potential for expansion, were captured by both light and electron microscopy on day 28 of cell culture. CONCLUSION: The reciprocal actions of NA and DMSO can control the proliferation of HPN-stimulated hepatocytes, which can be used for studying human hepatocyte metabolism, cytotoxicity, biotransformation and mutagenesis, and may provide experimental evidences for the treatment of liver failure and genetic liver diseases with in vitro hepatocyte clones. PMID- 12581954 TI - [Effect of liposome on permeability and drug retention of sodium fluorescein in rat skin: an experimental study]. AB - OBJECTIVE: To investigate the effect of liposome on the permeability and drug retention of sodium fluorescein(NaFl) in rat skin. METHODS: With an improved Franz diffusion cell and 0.125% NaFl liposome suspension as the model drug, in vitro measurement of percutaneous absorption and skin reservoir capacity for NaF1 was conducted using a fluorescence spectrophotometry at 4 and 12 h respectively after the diffusion experiment had started, and distribution of the fluorescence in rat skin was observed under fluorescence microscope at 4 h penetration experiment. NaFl solution of the same concentration as the model drugs served as the control for this experiment. RESULTS: In comparison with NaFl solution, liposomal NaFl suspension resulted in larger amount of NaFl retention in rat skin but smaller amount in the receiver cell in a four-hour Franz diffusion test, with also higher fluorescence intensity in the skin, especially the skin around the hair follicles. CONCLUSION: Local high drug concentrations can be achieved in the skin by liposomal suspension of water soluble drug for more effective treatment of skin diseases. PMID- 12581955 TI - [Tumor necrosis factor-alpha pretreatment for in vitro culture of mature dendritic cells]. AB - OBJECTIVE: To explore a new method for in vitro culture of mature dendritic cells (DCs) by utilizing the maturation-inducing effect of tumor necrosis factor alpha(TNF-alpha) on DC precursors. METHODS: Freshly isolated DC precursors (peripheral blood-derived monocytes) were initially cultivated for 4 h in RPMI 1640 supplemented with 10% fetal bovine serum and containing TNF-alpha (200 U/ml), followed by treatment with rhGM-CSF (800 U/ml) and interleukin (IL)-4 (500 U/ml) added into the medium. After 48 and 96 h, the 3 cytokines were simultaneously added to the medium for further culture. On day 8 of cell culture, the DCs were harvested and designated as T-DCs, which were subsequently subjected to examinations of flow cytometry analysis, assay by autologous and allogeneic mixed lymphocyte reaction (MLR) for estimating their maturation, and MTT assay for their specific killing activity. RESULTS: The T-DCs expressed specific surface markers (CD83,CD1a, CD54, CD11a, CD80, CD86, HLA-DR, CD83HLA-DR) of mature DCs, and stimulated strong T-cell proliferative responses in both autologous and autogeneic MLR. It also activated CD4+ T cells, which proliferated in response to autologous tumor, and specifically lysed tumor targets. CONCLUSION: The DCs thus cultured in vitro possess typical phenotype, and can present antigens to T cells, and this method provides an alternative in culturing mature and functionally competent DCs. PMID- 12581956 TI - [Screening of mimotope of Salmonella typhimurium lipopolysaccharide from phage displayed peptide]. AB - OBJECTIVE: To identify and characterize the mimotope of lipopolysaccharide (LPS) from cyclic 7-mer phage peptide library. METHODS: Cyclic 7-mer phage-displayed peptide library was screened using monoclonal antibody 2F4 (mAb 2F4) against Salmonella typhimurium LPS as the target, and the selected clones were tested by sandwich enzyme-linked immunosorbent assay (ELISA) and specific antigen inhibition ELISA. RESULTS: After 3 round of screening, 34 of the 38 selected clones were identified as positive for binding to mAb 2F4. Salmonella typhimurium LPS was capable of inhibiting the binding between the cones and mAb 2F4, with the 50% inhibitory concentration of all the positive clones within the range of 0.125 0.250 microg/ml. Sequence analysis was performed for 10 of the positive clones, whose amino acid sequences were subsequently deduced, and 7 of them had conservative amino acid of P-X-WAS-X-W with the mean hydrophobic amino acid content of 71.4% in all the sequences. CONCLUSION: The phage-displayed peptide is capable of simulating the epitope of Salmonella typhimurium LPS. PMID- 12581957 TI - [Clinical study of changes of urotensin II in patients with congestive heart failure]. AB - OBJECTIVE: To investigate the changes of plasma urotensinII (UII) levels in patients with congestive heart failure (CHF) of different severities and their clinical implications. METHODS: Plasma UII was determined by radioimmunoassay in 45 patients with CHF and 20 healthies control subjects. In all the subjects, the left ventricular fraction (LVEF) and the ratio of E/A were measured by echocardiography. RESULTS: The plasma UII level was significantly lowered in CHF patients in comparison with that in control subjects (1.41+/-1.09 pg/ml vs 4.35+/ 1.22 pg/ml, P=0.000). A significant correlation of plasma UII level with LVEF (r=0.540, P=0.000) and the E/A ratio was observed (r=0.539, P=0.000), and the severity of CHF was shown to be inversely correlated with plasma UII levels in the patients (r=-0.656, P=0.000), which was elevated after treatment. CONCLUSION: UII might play a role in the pathophysiological process of CHF, and its plasma level may serve as an indicator of the severity of CHF. PMID- 12581958 TI - [Two restriction fluorescence labeling methods for enhancing the signal-to-noise ratio of cDNA microarray hybridization]. AB - OBJECTIVE: To study the signal-to-noise ratio (SNR) of two restricted fluorescence labeling methods for examining gene expression profile by microarray hybridization. METHOD: Samples of Saccharomyces cerevisiae mRNA was labeled by traditional reverse transcription method and 2 restriction fluorescent labeling methods using respectively Cy-universal primer and extension incorporated Cy dNTP. The labeled samples were examined by the microarray, followed by washing and scanning under the same conditions. RESULTS: The two restriction labeling methods showed superior results with lowered background and enhanced SNR and sensitivity, and Cy-universal primer labeling presented the best results. CONCLUSION: SNR can be enhanced by the restriction labeling methods, which improve the applicability of microarray technology. PMID- 12581959 TI - [Establishment of transgenic mouse models carrying human polymeric immunoglobulin receptor gene]. AB - OBJECTIVE: To establish transgenic mouse models carrying human polymeric immunoglobulin receptor (pIgR) gene to render the mice prone to EBV infection in normal condition, thereby to facilitate the study of the role of Epstein-Barr virus (EBV) in the pathogenesis of nasopharyngeal carcinoma. METHOD: By means of microinjection, pIgR gene under the regulation by kerotinocyte-specific promoter ED-L2 was introduced into the pronuclei of mouse zygote, which was transferred into pseudo- pregnant female mice to induce nasopharynx-specific pIgR expression in the founder mice. RESULT: Six out of the 16 founder mice (37.5%) was tested by PCR to be pIgR-positive, while Southern blotting identified only 2 positive mice. CONCLUSION: Transgenic mice harboring pIgR gene can be used potentially as a new model for studying the pothogenesis of nasopharyngeal carcinoma by EBV. PMID- 12581960 TI - [In vitro biocompatibility and degradation of nacre/polylactic acid composite artificial bone]. AB - OBJECTIVE: To investigate the in vitro biocompatibility and degradation of nacre/polylactic acid composite artificial bone (NPCB) grafts. METHODS: Human osteoblast cells were cultured with NPCB discs for observation of the morphological features and cell proliferation by optical microscope and scanning electron microscope. The mass loss of NPCB discs and pH variations of the saline in which the discs were immersed were examined every 2 weeks in a course of 16 weeks in vitro. RESULT: Compared with the negative control group, NPCB showed no visible cytotoxicity and facilitated the growth and proliferation of the osteoblast cells. As the immersion prolonged, gradual decrease in the mass of NPCB was observed accompanied by regular pH alteration of the saline. CONCLUSION: NPCB possesses good in vitro biocompatibility and may be gradually degraded. PMID- 12581961 TI - [Nonlinear dynamic analysis of heart rate variability in patients with diabetic autonomic neuropathy]. AB - OBJECTIVE: To explore earlier detection methods for diabetic autonomic neuropathy (DAN) by heart rate variability (HRV) analysis. METHODS: Thirty-four diabetic patients (including 22 with explicit clinical DAN symptoms) were randomly selected for this study from the in-patient department of endocrinology. On the basis of Virtual Instrumental WorkBench- LabVIEW and using several nonlinear dynamic analysis measures, including Allan factor, lyapunov exponent, approximate entropy, fractal dimension, complexity, wavelet-transform standard deviation and nonlinear energy operator, the analysis of the HRV in these diabetic patients was performed in comparison with normal subjects. RESULTS: The nonlinear indices of both DAN patients and patients without obvious DAN were significantly different from those of the normal subjects, especially in terms of Lyapunov exponent, approximate entropy, and nonlinear energy operator. CONCLUSION: Nonlinear dynamic methods of HRV analysis can provide assistance in assessing the status and impairment of the autonomic system, and can be used to efficiently detect diabetic neuropathy in early stages. PMID- 12581962 TI - [In vitro culture of human hair follicles and sequential observation]. AB - OBJECTIVE: To obtain successful in vitro culture of human hair follicles, and observe their morphological changes in the middle and later period of culture. METHODS: Human hair follicles were isolated from the scalp resected from patients receiving surgical treatment of the wrinkles, and were cultured in DMEM supplemented with 10% calf serum for up to 30 d. The growth of the follicles, along with their morphological and histological changes, was observed sequentially. RESULTS: In vitro culture of human hair follicles was successfully obtained. The follicles showed catagen-like changes in the middle and later period of culture. CONCLUSION: Human hair follicles cultured in vitro tend to transform early into catagen, and this culture method has simplified preparation of routine paraffin section of hair follicles. PMID- 12581963 TI - [Morphological study on three-dimensional culture of rat muscle satellite cells under cyclic mechanical stretch]. AB - OBJECTIVE: To study the effect of mechanical stretch on the morphologic change of rat muscle satellite cell in the three-dimensional culture. METHODS: Based on the apparatus for three-dimensional cell cultures under a cyclic mechanical strain, a specific stretch pattern (10% elongation, 10 stretches/min for 10 min of each hour 48 h total) was applied to cell-scaffolding composites. RESULTS: Under the stretch pattern, the shape of satellite cells changed to oblate and spread to the direction of the stretch. Furthermore, the myotube was observed. On the contrary, the satellite cells spread to the all direction, and the formation of the myotube was not been found in the control group. CONCLUSION: Cyclic mechanical stretch is helpful for the formation of the ideal directivity and these results are compatible with a significant role for the stretch in tissue-engineered muscle construction. PMID- 12581964 TI - [Effect of antisense oligodeoxynucleotides of RNA component of human telomerase on cisplatin induced apoptosis in K562 cells]. AB - OBJECTIVE: To research whether the antisense oligodeoxynucleotide (ASODN) of RNA component of human telomerase (hTR) can increase the susceptibility of K562 cells to cisplatin. METHODS: K562 cells were treated with phosphorothoate ASODN complementary to the template region of hTR in vivo. The changing of telomerase activity was assayed by TRAP-PCR-ELISA and apoptosis by DNA gel electrophoresis, Annexin V, flow cytometry. RESULTS: There was a marked decrease in telomerase activity in ASODN treated cells as compared with that in control and sense oligodeoxynucleotide (SODN)-treated cells; but no difference between the latter two groups. Agarose gel electrophoresis of genomic DNA from K562 cells treated with ASODN and cisplatin combination for 72 h showed typical DNA ladder; neither did DNA ladder from K562 cells treated with SODN plus cisplatin nor cisplatin alone. Apoptosis rates of K562 cells treated with ASODN for 24 h and then with cisplatin for 72 h were significantly increased. There were statistically significant difference in the percentage of apoptotic cells between hTR ASODN plus cisplatin and SODN plus cisplatin or cisplatin alone group. CONCLUSION: ASODN complementary to the template region of hTR can significantly suppress the telomerase activity and enhance cisplatin-induced apoptosis of K562 cells in vivo. PMID- 12581965 TI - [The effect of bending and rotation on the lumbar facet joints under load-bearing conditions]. AB - OBJECTIVE: To evaluate the effect of bending and rotation on the lumbar facet joints under load-bearing conditions. METHODS: Eight lumbar-sacral spines (L1-S1) taken from fresh young adult cadavers were placed on the MTS system. The forces of the two facet joints of the L4-5 motion segments were measured using pressure sensitive films. The specimens were measured in erection, bending and rotation postures. RESULTS: In erection posture the L4-5 facet joints could support 31.16+/-7.15 N of 400 N which was axial compressing load. In bending 15 degree posture the forces of each side of the L4-5 facet joints were equal, and were found to have no marked effect on the force in erection. Yeat only opposite facet joints could endure force in rotation, but the force was 7 times that in erection. CONCLUSION: Both sides of lumbar facet joints could support load in bending position but only the opposite could endure load in rotation. PMID- 12581966 TI - [Effects of three different immunosuppressive drugs on SD rat islet cell viability]. AB - OBJECTIVE: To investigate the effect of immunosuppressive drugs on the viability of pancreatic islet cells cultured in vitro, and to explore their effects on the outcome of islet transplantation. METHODS: Pancreatic islet cells from SD rats were cultured in vitro, and the changes in their viability determined by MTT assay after incubation with immunosuppressive drugs at different concentrations. RESULTS: The viability of the islet cells was reduced after exposure to rapamycin at higher concentrations (>or= 1 ng/ml), but not at lower concentrations. Treatment with daclizumab and FTY720, at either lower or higher concentrations, did not result in significant reduction of the islet viability. CONCLUSIONS: Rapamycin may directly cause damage to islet cells, the degree of which is positively correlated to the concentration of rapamycin. No obvious toxicity of daclizumab and FTY720 was observed against the islet cells. PMID- 12581967 TI - [A new method for detecting serum beta-lipoprotein electrophoretogram]. AB - OBJECTIVE: To establish a new method for isolating serum beta-lipoprotein (betaLp) subfractions using second double gradient polyacrylamide gel electrophoresis (s-DGPAGE). METHODS: After the betaLp subfractions were isolated by mechanisms of molecular sieving and isoelectric focusing due to density and pH grade in the non-linear density gradient polyacrylamide gel, the gels were acquired with a gel imaging system, the electrophoretograms were quantitatively analyzed with optical density scanner. The isolation of serum betaLp was completed through first DGPAGE. RESULTS: s-DGPAGE separated the serum betaLp into four subfractions, designated respectively as betaLp1, betaLp2, betaLp3 and betaLp4, and their amounts in the betaLp and in the whole lipoproteins could be quantified. CONCLUSION: s-DGPAGE is a simple and practical method to show the heterogeneity of serum betaLp and the quantitative variation of its subfractions. PMID- 12581968 TI - [Association of platelet endothelial cell adhesion molecule-1 gene polymorphism with coronary heart disease]. AB - OBJECTIVE: To investigate the association of Leu125Val and Ser563Asn polymorphism of the gene encoding platelet endothelial cell adhesion molecule-1(PECAM-1) with coronary heart disease. METHODS: This study included 156 patients with the diagnoses of coronary heart disease (CHD) and coronary lesions derived from electrocardiography, myocardial enzyme analysis and coronary angiography as the CHD group, and another 75 in-patients admitted within the same period who showed no signs of CHD in the above examinations constituted the control group. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed to examine the missense polymorphism of PECAM-1gene in the position of Leu125Val and Ser563Asn. RESULTS: There were significant differences between CHD and control group in terms of the allele frequencies and genotype distributions of PECAM-1 gene, and the differences were especially conspicuous in the allele frequencies of 125Val and 563Asn (P<0.05) and genotype distributions of 125Val/Val and 563Asn/Asn. CONCLUSION: PECAM-1 gene polymorphism 1 may be a genetic risk factor for coronary heart disease. PMID- 12581969 TI - [Clinical value of 18F-fluoro-2-deoxy-D-glucose positron emission tomography in detecting recurrent nasopharyngeal carcinoma at the parapharyngeal space after radiotherapy]. AB - OBJECTIVE: To explore the significance of 18F-fluoro-2-deoxy-D-glucose positron emission tomography (18FDG-PET) in the diagnosis of the recurrence or fibrosis of nasopharyngeal carcinoma at the parapharyngeal space after radiotherapy. METHODS: Thirty-two patients having received radiotherapy for nasopharyngeal carcinoma at the parapharyngeal space underwent 18FDG-PET in parallel with CT and magnetic resonance imaging (MRI) examinations to detect the recurrence or fibrosis of the tumors after the treatment, when the latter two examinations had failed to yield definite results. After all the imaging examinations, CT-guided puncture biopsy was performed in each case to observe the sensitivity, specificity and accuracy of the examinations. RESULTS: The sensitivity, specificity and accuracy were 79.16%, 75.00%, and 78.13% respectively for CT examination, and were 83.33%, 87.50%, and 84.38% for MRI. 18FDG-PET had the best sensitivity, specificity and accuracy in detecting tumor recurrence, which were 95.83%, 87.50%, and 93.75% respectively. CONCLUSION: 18FDG-PET can be considered in cases of suspected recurrence of nasopharyngeal carcinoma at the parapharyngeal space after radiotherapy, and CT and MRI examinations either used alone or in combination may also reach definite diagnosis with the help of CT-guided biopsy. PMID- 12581970 TI - [Expression of tyrosine kinase receptor B in human follicle]. AB - OBJECTIVE: To study the expression of tyrosine kinase receptor B(TrkB) in human follicle and explore the significance of the expression. METHODS: Streptavidin biotin peroxidase complex immunocytochemistry was employed to determine the expression of TrkB on the oocytes, cumulus granulosa cells (CGCs) or mural granulosa cells (MGCs) obtained from women during controlled ovarian hyperstimulation for in vitro fertilization-embryo transfer (IVF-ET). Western blotting was used to analyze TrkB expression level in CGCs and MGCs. RESULTS: The results demonstrated for the first time the presence of TrkB on the surface of human ovary cells including MGCs, CGCs and the oocytes. Higher expression levels of TrkB were detected on the MGCs than on the CGCs. CONCLUSION: It is possible that TrkB tyrosine autophosphorylation in response to neurotrophins can take part in the regulation and modulation of steroidogenesis and oocyte maturation. PMID- 12581971 TI - [Diagnostic value of combined detection of TPS, NSE and CEA in lung cancer]. AB - OBJECTIVE: To study the diagnostic value of combined detection of 3 tumor markers, namely tissue polypeptide specific antigen (TPS), neuro-specific enolase (NSE) and carcinoembryonic antigen (CEA), in patients with lung cancer. METHODS: The serum levels of TPS, NSE and CEA were determined by enzyme-linked immumosorbent assay in 72 patients with lung cancer and 114 healthy adults. RESULTS: The levels of the 3 tumor markers in the patient group were significantly higher than those of the healthy control group (P<0.001). The sensitivity of TPS for lung cancer detection was 84.7% with specificity of 89.4%, which were 54.1% and 91.2% respectively for NSE, and 45.8% and 91.4% respectively for CEA. The combined detection of the 3 tumor markers for lung cancer diagnosis had a sensitivity of 100%. CONCLUSIONS: TPS, NSE and CEA employed separately are helpful in the diagnosis of lung cancer, and their combined detection may significantly improve the sensitivity for lung cancer detection, due to the obvious complementarity of the 3 markers. PMID- 12581972 TI - [Ropivacaine or its combination with fentanyl for postoperative patient controlled epidural analgesia]. AB - OBJECTIVE: To investigate the pain-relieving effects of ropivacaine or its combination with fentanyl in postoperative patient-controlled epidural analgesia. METHODS: Sixty patients (ASA class I to II) scheduled for elective upper abdominal surgery were randomly divided into 4 groups (15 in each group) to receive 2.0 mg/ml ropivacaine (group R), 1.25 mg/ml ropivacaine plus 1.0 microg/ml fentanyl (group RF1), 1.0 mg/ml ropivacaine plus 2.0 microg/ml fentanyl (group RF2), and 1.0 mg/ml ropivacaine plus 4.0 microg/ml fentanyl (group RF4) respectively for patient-controlled analgesia. The bolus injection was set at 3.0 ml, lockout interval at 20 min, and background epidural infusion at 5.0 ml/h. Before and at 0, 6, 12, 24 and 48 h after the surgeries, the data including mean arterial pressure (MAP), heart rate (HR), cumulative consumption volume (CCV) of analgesic solution and visual analog scale (VAS) scores were recorded, observed and the adverse effects such as vomiting, nausea, pruritus, as well as the time of first excretion of intestinal gas. RESULTS: Compared with group R, significantly smaller CCV occurred in groups RF2, RF4 (P<0.01), and RF1 (P<0.05) at each time points postoperatively. No significant differences were noted between the groups in VAS scores. CONCLUSION: The 4 preparations are comparable in terms of analgesic efficacy and adverse effects, but in groups RF2 and RF4, the smaller doses have sufficed the needs. PMID- 12581973 TI - [Value of serum progesterone for ectopic pregnancy in therapeutic effect monitoring]. AB - OBJECTIVE: To study the value of serum progesterone in selecting the candidate patients with ectopic pregnancy (EP) for methotrexate (MTX) treatment and in monitoring the effect of the treatment. METHODS: Thirty-seven EP patients who were given single-dose intramuscular injection with 50 mg/m2 MTX were divided into success and failure groups according to the effect of the treatment. The serum progesterone concentrations in these patients were measured and compared between the 2 groups, and the receiver-operator curves (ROC) were used to determine the critical serum progesterone levels for assessing the effect of MTX treatment. The time consumed respectively by serum progesterone and beta-human chorionic gonadotrophin (beta-hCG) decreasing to below normal level was also compared. RESULTS: The serum progesterone concentrations of the success group (7.93+/-2.02 ng/ml) were significantly lower than those of the failure group (14.53+/-1.72 ng/ml, P<0.05). The recommended critical level for assessing the effect of MTX treatment for EP patients was 11 ng/ml according to the ROC, and the time for serum progesterone decreasing to below normal level was significantly less than that for beta-hCG. CONCLUSION: Serum progesterone can be used as an index for selecting candidate EP patients for MTX treatment, and also as a good indicator for assessing the therapeutic effect after treatment. PMID- 12581974 TI - [Glucocortioid treatment for heroin-induced spongiform leucoencephalopathy: a clinical controlled study]. AB - OBJECTIVE: To evaluate the therapeutic effects of the glucocorticoid on heroin induced spongiform leucoencephalopathy. METHODS: Twenty cases of heroin-induced spongiform leucoencephalopathy were randomly divided into the control group and the treating group with equal number. In the control group, the treatment was constituted by oral administration of vitamin B and coenzyme Q10 in a course of 1 month. In glucocorticoid treatment group, glucocorticoid (20 mg/d) for 10 d were given in addition to vitamin B and coenzyme Q10, and the dose of the glucocorticoid was gradually decreased afterwards. General observation and statistical analysis of function scores were performed in both groups before and 1, 6, 12 months after the treatment respectively. RESULTS: No significant difference in function scores was observed between the 2 groups, while the results of observation before and after the treatment were significantly different (P<0.05). The most significant difference occurred when comparing the observations made 1 month and 6 months respectively after treatment (P<0.001). CONCLUSION: Glucocorticoid has no obvious therapeutic effect on heroin-induced spongiform leucoencephalopathy, and rapid clinical recovery occurs within the initial 6 months of the treatment. PMID- 12581975 TI - [Retrospective study of 10 cases of transplant renal artery stenosis]. AB - OBJECTIVE: To investigate the incidence and cause of transplant renal artery stenosis (TRAS) and explore its diagnosis and management. METHOD: A retrospective study was conducted in 10 TRAS cases confirmed by selective artery angiography between 2000 and 2001. RESULTS: Seven patients had a history of acute rejection episode, balloon angioplasty was per-formed in 9 patients, and stent implantation in 1 patient. Revascularization resulted in a decrease in arterial pressure and renal function improvement in all but 1 case. Restenosis occurred in 2 of the 9 patients receiving balloon angioplasty, but after stent implantation, their renal function was improved. Serious graft failure occurred in 1 case, resulting eventually in renal graft loss. CONCLUSIONS: TRAS is related to episodes of acute rejection. Doppler ultrasonography is useful in screening of TRAS, the definite diagnosis of which relies on artery angiography, and angioplasty constitutes an effective therapeutic modality. PMID- 12581976 TI - [Diagnosis and treatment of traumatic tracheobronchial ruptures: report of 17 cases]. AB - OBJECTIVE: To evaluate the diagnosis and treatment of tracheobronchial ruptures due to thoracic trauma. METHODS: The clinical data of 17 cases of bronchial rupture caused by chest trauma was analyzed retrospectively. The surgical approaches and post-trauma complications were described. RESULTS: End-to-end anastomosis was performed in 14 cases. Among the other 3 patients, one received lobectomy after bronchial repair, another had tracheal repair, and resection of the stenosed segment along with end-to-end anastomosis during thoracotomy was necessitated in the last case. Anastomotic stricture occurred early after the operation in 2 of all these cases, while 2 months postoperatively, all the patients were free from such strictures. CONCLUSION: Early diagnosis and operation in such cases can be life-saving, and may warrant early recovery of the pulmonary function. Accurate diagnoses of bronchial rupture can be obtained by bronchoscopy, with which the surgeons can easily locate the rupture during surgery. PMID- 12581977 TI - [Diagnosis and surgical treatment of primary cardiac tumors]. AB - OBJECTIVE: To study the clinical characteristics, diagnosis, and surgical treatment of primary cardiac tumors. METHOD: The clinical features and surgical results in 126 cases (1971-2000) of primary cardiac tumors were analyzed retrospectively. RESULTS: Among the 121 patients with benign tumors were treated surgically, 6 had recurrence of the pathologically confirmed intracardiac myxoma in a period of 1 to 7 years after surgery, and all were cured after reoperation. Follow-up of these patients showed satisfying long-term survival. In the other 5 malignant cases, 1 died during the perioperative period, and the others died of progressive heart failure, cardiac tamponade and cardiac rupture, respectively, within 10 months after the final diagnosis. CONCLUSION: The symptoms of primary cardiac tumor are diverse and complex, and echocardiography, CT, MRI are instrumental for a definite early diagnosis. In case of benign tumors, operation usually produces good results, but for malignant tumors characterized by rapid progression, earlier surgical removal in combination with other therapeutic modalities may improve the prognosis. PMID- 12581978 TI - [Imaging diagnosis of xanthogranulomatous pyelonephritis: report of 6 cases]. AB - OBJECTIVE: To explore the imaging diagnosis of xanthogranulomatous pyelonephritis (XGP), especially the value of angiography. METHODS: Combined with literature review, a retrospective analysis was conducted of the imaging data of 6 pathologically confirmed XGP cases as an attempt to define the preoperative diagnosis of the disease. RESULTS: Four cases of XGP were diagnosed preoperatively, and 2 were misdiagnosed as renal abscess. Renal arteriography revealed more characteristic changes, and the diagnosis of XGP relied on the clinical presentations and multiple imaging examinations. CONCLUSION: Renal arteriography is the principal approach in the diagnosis and differential diagnosis of XGP. PMID- 12581980 TI - [Early diagnosis and treatment of intracranial venous thrombosis]. AB - OBJECTIVE: To study the early diagnosis and treatment of intracranial venous thrombosis. METHOD: The etiology, clinical manifestations, presentations in cerebral CT and magnetic resonance imaging (MRI), electroencephalogram (EEG) features, and the results of cerebral spinal fluid and blood rheological examinations were analyzed in 8 cases of intracranial venous thrombosis. RESULTS: Thrombosis often arose during the puerperium and from infection, with the frequent presentations of intracranial hypertension and focal neurological defect. CT and MRI examinations were helpful in the diagnosis. CONCLUSION: For patients who present both intracranial hypertension and focal neurological defects, especially during the puerperium or with infection episodes, CT and MRI examinations should be performed to detect intracranial venous thrombosis. PMID- 12581979 TI - [Clinical analysis of 410 cases of drug eruption]. AB - An clinical analysis was conducted among a cohort of 410 patients drug eruption with treated in our department from January 1995 to December 2001. We found that the common drugs likely to lead to anaphylactic reactions included cephalosporins, ampicillin types, antipyretic analgesic types, rabies vaccine, sulfonamides, tetracyclines types, etc. The drug eruption mostly presents diverse clinical features resembling the rashes as seen in cases of scarlet fever, measles, urtica, or mucosal edema or ulceration. PMID- 12581981 TI - [All-trans retinoic acid for thromboembolic events in patients with M3 leukemia: a case report]. AB - A case of acute promyelocytic leukemia (APL) treated with all-trans retinoic acid (ATRA) is reported. After the diagnosis was established, the patient was given oral ATRA (30 mg/d) in combination with small dose of hydroxyurea, platelet concentrates, and fresh frozen plasma etc. From day 19 after ATRA administration, successive thromboembolic events occurred. In spite of the partial remission on day 32, the patient died of cardiopulmonary insufficiency. Our experience from this case suggests that more attention should be given to thromboembolic events during ATRA therapy, and the use of active anti-coagulant may prove beneficial when signs of hypercoagulation are present in APL patients. PMID- 12581982 TI - [Clinical analysis of 22 cases of Bartholin gland cyst or abscess treated with trocar puncture]. AB - This study was designed to observe the effects of trocar puncture in the treatment of Bartholin gland cyst or abscess. The cure rate of these 22 cases was 100%, and no relapse of the disease occurred during the follow-up lasting for 5 to 12 months. Trocar puncture proves to be effective in the treatment of bartholin gland cyst or abscess, especially when the size is less than 3.0 cm. PMID- 12581984 TI - Control of Three- and Four-Joint Arm Movement: Strategies for a Manipulator With Redundant Degrees of Freedom. AB - Control of arm movements when the number of joints exceeds the degrees of freedom necessary for the task requires a strategy for selecting specific arm configurations out of an infinite number of possibilities. This report reviews strategies used by human subjects to control the shoulder, elbow, and wrist (three degrees of freedom) while moving a pointer to positions in a horizontal plane (two degrees of freedom). Analysis of final arm configurations assumed when the pointer was at the target showed the following: (a) Final arm configurations were virtually independent of the configuration at the start of the pointing movement, (b) subjects avoided configurations subjectively felt to be uncomfortable (e.g., those with extreme flexion or extension of the wrist), and (c) the results could be simulated by assigning hypothetical cost functions to each joint and selecting the arm configuration that minimized the sum of the costs. The fitted cost functions qualitatively agreed with psychophysically determined comfort; they appeared to depend on joint angle and on muscular effort. Simple neural networks can learn implicit representations of these cost functions and use them to specify final arm configurations. The minimum cost principle can be extended to movements that use the fingers as a fourth movable segment. For this condition, however, experiments showed that final configurations of the arm depended upon initial configurations. Analysis of movement trajectories for arms with three degrees of freedom led to a control model in which the minimum cost principle is augmented by a mechanism that distributes required joint movements economically among the three joints and a mechanism that implements a degree of mass-spring control. PMID- 12581985 TI - Control of Trajectory Modifications in Target-Directed Reaching. AB - Human reaching movements to fixed and displaced visual targets were recorded and compared with simulated movements generated by using a two-joint arm model based on the equilibrium-point (EP) hypothesis (lambda model) of motor control (Feldman, 1986). The aim was to investigate the form of central control signals underlying these movements. According to this hypothesis, movements result from changes in control variables that shift the equilibrium position (EP) of the arm. At any time, muscle activations and forces will depend on the difference between the arm's EP and its actual position and on the limb's velocity. In this article, we suggest that the direction of EP shift in reaching is specified at the hand level, whereas the rate of EP shift may be specified at the hand or joint level. A common mechanism underlying reaching to fixed and displaced targets is proposed whereby the EP of the hand shifts in a straight line toward the present target. After the target is displaced, the direction of the hand EP shift is modified toward the second target. The results suggest that the rate of shift of the hand EP may be modified for movements in different parts of the work space. The model, with control signals that vary in a simple fashion over time, is able to generate the kinematic patterns observed empirically. PMID- 12581983 TI - [Frequent mistakes in English medical papers (I)]. AB - In this article, we attempt to grammatically classify the mistakes that frequently appear in the English medical papers written by Chinese, for the purpose of drawing attention on these avoidable mistakes from the authors to improve their medical paper writing in English. PMID- 12581986 TI - A Computational Model of the Simplest Motor Program. AB - A computational procedure (program) is defined to generate control signals for the motoneuron pools of agonist and antagonist muscles that will move a limb segment from one stationary position to another. The program accounts for the ability to move different distances with different inertial loads and for the influence of instructions concerning movement speed and accuracy. These motor commands allow the program to produce EMG patterns as well as force and kinematic trajectories that are consistent with much of the data found in the literature of these movements. The program is premised on the notion that kinematically defined tasks are accomplished by programming commands to the motoneuron pools, based on only a few cognitively recognized kinematic and dynamic features of the task. Most of the features found in EMG and kinematic patterns can be considered consequences of the program's algorithmic procedures rather than specifically planned features of those movements. PMID- 12581987 TI - The Cascade Neural Network Model and a Speed-Accuracy Trade-Off of Arm Movement. AB - We propose a hybrid neural network model of aimed arm movements that consists of a feedforward controller and a postural controller. The cascade neural network of Kawato, Maeda, Uno, and Suzuki (1990) was employed as a computational implementation of the feedforward controller. This network computes feedforward motor commands based on a minimum torque-change criterion. If the weighting parameter of the smoothness criterion is fixed and the number of relaxation iterations is rather small, the cascade model cannot calculate the exact torque, and the hand does not reach the desired target by using the feedforward control alone. Thus, one observes an error between the final position and the desired target location. By using a fixed weighting parameter value and a limited iteration number to simulate target-directed arm movements, we found that the cascade model generated a planning time-accuracy trade-off, and a quasi-power-law type of speed-accuracy trade-off. The model provides a candidate neural mechanism to explain the stochastic variability of the time course of the feedforward motor command. Our approach also accounts for several invariant features of multijoint arm trajectories, such as roughly straight hand paths and bell-shaped speed profiles. PMID- 12581988 TI - Models of Trajectory Formation and Temporal Interaction of Reach and Grasp. AB - Our goal was to create a principled account of a body of behavioral kinematic data on reaching and grasping. We show how to transform an optimality principle for overall hand transport into a feedback control law and then incorporate look ahead modules in the controller to compensate for delays in sensory feedback. This model describes the kinematics of hand transport under a variety of circumstances, including target perturbations. We then develop a model for the temporal coordination of reach and grasp. We provide an optimization principle for hand preshaping that trades off the costs of maintaining the hand in an open position and the cost of accelerating the change in grip size. This yields a control system for preshaping. We then show that a model that uses only expected duration for coordination, rather than kinematic or dynamic variables, can describe the kinematics of interaction of hand transport and preshape under a variety of circumstances, including perturbations of object position and object size. PMID- 12581989 TI - Servo Hypotheses for the Biological Control of Movement. AB - An analysis is made of equilibrium-point models for motor control, describing these models in the context of servo control mechanisms. We considered issues of speed and stiffness scaling that are incompatible with current formulations of the equilibrium-point models. A modification of the equilibrium-point models is proposed in which the central nervous system controls velocity as well as positions during the course of fast 1imb movements. Numerical simulations are presented that verify that such a servo control mechanism could successfully produce fast limb movements, as observed in human subjects PMID- 12581990 TI - Is the cerebellum a smith predictor? AB - The motor system may use internal predictive models of the motor apparatus to achieve better control than would be possible by negative feedback. Several theories have proposed that the cerebellum may form these predictive representations. In this article, we review these theories and try to unify them by reference to an engineering control model known as a Smith Predictor. We suggest that the cerebellum forms two types of internal model. One model is a forward predictive model of the motor apparatus (e.g., limb and muscle), providing a rapid prediction of the sensory consequences of each movement. The second model is of the time delays in the control loop (due to receptor and effector delays, axonal conductances, and cognitive processing delays). This model delays a copy of the rapid prediction so that it can be compared in temporal register with actual sensory feedback from the movement. The result of this comparison is used both to correct for errors in performance and as a training signal to learn the first model. We discuss evidence that the cerebellum could form both of these models and suggest that the cerebellum may hold at least two separate Smith Predictors. One, in the lateral cerebellum, would predict the movement outcome in visual, egocentric, or peripersonal coordinates. Another, in the intermediate cerebellum, would predict the consequences in motor coordinates. Generalization of the Smith Predictor theory is discussed in light of cerebellar involvement in nonmotor control systems, including autonomic functions and cognition. PMID- 12581991 TI - Knowledge Model for Selecting and Producing Reaching Movements. AB - This article presents a new model of reaching control. The aim of the model is to characterize the computations underlying the selection of coordinated motion patterns among the limb segments. When a spatial target is selected, stored postures are evaluated for the contributions they can make to the task, and a special weighted average (the gaussian average) is taken of the postures to find a single target posture. Movement to the target posture is achieved without explicit planning of the trajectory. Rather, the reaching motion is driven by error correction (reducing the discrepancy between the current and target posture) shaped by inertia. The model solves the degrees-of-freedom problem for reaching. It also allows joints to compensate automatically for reduced mobility of other joints and explains established effects of practice, speed-accuracy trade-off, and kinematics. The model can be extended to other tasks and motor subsystems because of the generality of its underlying concepts. PMID- 12581992 TI - Control of Equilibrium Position and Stiffness Through Postural Modules. AB - If muscles are viewed as spring-like torque generators, then the integral of torque with respect to joint angle is the potential energy of that muscle. An energy function for the musculoskeletal system can be defined by summing the energy contribution of each muscle and the potential energy stored in the limb. Any local minimum in this energy landscape is a possible equilibrium position for the limb. The gradient of this function with respect to joint angles is a torque field, and the task of postural control is to find a set of muscle activations to produce a desired field. We consider one technique by which this approximation may be achieved: A postural module is defined as a synergy of muscles that produce a class of torque functions that converge at a constant equilibrium position, but whose stiffness at this position varies as a function of activation of the postural module. For a single-joint system, we show that through control of two such modules it is possible to produce any stiffness at any desired equilibrium position. To extend this scheme to a multijoint system, we initially derive the mechanical constraints on the shape of the restoring force field when a multijoint limb is displaced from equilibrium. Next, we consider voluntary control of the force field when the human arm is displaced from equilibrium: Mussa-Ivaldi, Hogan, and Bizzi (1985) have suggested that subjects are unable to voluntarily change the shape and orientation of the field, although they can readily scale it. This suggests existence of a limitation on the independent recruitment of arm muscles. We show, through simulation, that the inability to voluntarily control the shape and orientation of the restoring force field can be attributed to an organization of postural modules that act as local stiffness controllers. We propose that through coactivation, postural modules coarsely encode the work space and serve as an intermediate control system in the motor control hierarchy. PMID- 12581993 TI - Are the Orientations of the Head and Arm Related During Pointing Movements? AB - The head, eye, and shoulder are each free to rotate around three mutually orthogonal axes. These three degrees of freedom allow a given gaze or pointing direction of the eye, head, or arm to be obtained in many different possible orientations. Unlike translations in three dimensions, three-dimensional (3-D) rotations are noncommutative. Therefore, the orientation of a rigid body following sequential rotations about two different axes depends on the order of the rotations. In this article, we demonstrate that only two degrees of freedom are used during orienting movements of the head and pointing movements of the arm. This provides a unique orientation of head and arm for each gaze or pointing direction despite the noncommutativity of three-dimensional rotations. This observation is in itself not new. We found, however, that (a) the two-dimensional lquot;rotation surface,rquot; which describes the orientation of the head for all gaze directions, is curved, unlike the analogous flat plane for the eye. (b) The rotation surface for the head is curved differently than that for the arm. This result argues against the hypothesis that the orientations of head and arm are directly coupled during pointing. It also implies that the orientation of the eye in space during gaze shifts of the eye and head is not uniquely determined for a given direction of gaze. This finding argues against a perceptual basis for the reduction of rotational degrees of freedom. PMID- 12581994 TI - Commentary: variability in workplace exposures and the design of efficient measurement and control strategies. AB - Kromhout et al.'s (1993) well-cited publication presented detailed information on statistical procedures to estimate the magnitude of exposure variability within and between workers, drawing from a large database on chemical exposures throughout industry. It convincingly demonstrated that the construct of homogeneous exposure groups often does not hold true and suggested ways to improve measurement strategies. The authors hit a rich vein of research, and many publications, not at least by the authors themselves, followed in the decade after publication. In recent years the principles of estimating the variation in exposure have been applied in new methods for optimization of sampling strategies, for compliance testing, for quantifying exposures in epidemiologic studies, and for identifying important sources of emissions and suggesting strategies for controlling exposures. Many occupational hygienists across the globe have adopted these new methods as powerful tools in their exposure assessment strategies. PMID- 12581995 TI - Comparison of measurement strategies for prospective occupational epidemiology. AB - In the context of a prospective assessment of exposure for epidemiology, our objective is to obtain an optimal group-based design of allocation of a fixed total number of measurements. Such a design has been described by Ashford [Ashford JR. (1958) The design of a long-term sampling programme to measure the hazard associated with an industrial environment. J R Statist Soc A; 121: 331 47]. As this strategy is not operational, we developed three series of strategies: the first based on simplifications of Ashford's strategy; the second based on a pilot study; and the third on an iterative assessment of the group specific standard deviation of exposure. These strategies are compared by simulating a day-to-day individual exposure in several industrial sites and the resulting health effect. Our criteria for comparing strategies are the mean squared error of the estimated exposure in each group weighted by the number of subjects and the mean squared error of the estimated linear regression coefficient in the dose-response relationship. Strategies relying on an iterative approach have been found to perform best whatever the circumstances, nearly as well as Ashford's optimal strategy. PMID- 12581996 TI - Excessive exposure to silica in the US construction industry. AB - Exposures to respirable dust and silica were investigated among 36 construction sites in the USA. Personal measurements (n = 151) were analyzed from 80 workers in four trades, namely bricklayers, painters (while abrasive blasting), operating engineers and laborers. Painters had the highest exposures (median values for respirable dust and silica: 13.5 and 1.28 mg/m(3), respectively), followed by laborers (2.46 and 0.350 mg/m(3)), bricklayers (2.13 and 3.20 mg/m(3)) and operating engineers (0.720 and 0.075 mg/m(3)). Mixed models were fitted to the log-transformed air levels to estimate the means and within- and between-worker variance components of the distributions in each trade. We refer to the likelihood that a typical worker from a given trade would be exposed, on average, above the occupational exposure limit (OEL) as the probability of overexposure. Given US OELs of 0.05 mg/m(3) for respirable silica and 3 mg/m(3) for respirable dust, we estimated probabilities of overexposure as between 64.5 and 100% for silica and between 8.2 and 89.2% for dust; in no instance could it be inferred with certainty that this probability was <10%. This indicates that silica exposures are grossly unacceptable in the US construction industry. While engineering and administrative interventions are needed to reduce overall air levels, the heterogeneous exposures among members of each trade suggest that controls should focus, in part, upon the individual sites, activities and equipment involved. The effects of current controls and workplace characteristics upon silica exposures were investigated among operating engineers and laborers. Silica exposures were significantly reduced by wet dust suppression (approximately 3-fold for laborers) and use of ventilated cabs (approximately 6 fold for operating engineers) and were significantly increased indoors (about 4 fold for laborers). It is concluded that urgent action is required to reduce silica exposures in the US construction industry. PMID- 12581997 TI - Estimating aerosol surface area from number and mass concentration measurements. AB - A number of toxicology studies have been published indicating that health effects associated with low-solubility inhaled particles may be more appropriately associated with particulate surface area than mass. While exposure data from the workplace is needed to further investigate the relevance of such an association, the means of measuring exposure to aerosol surface area are not readily available. A possible interim solution is to estimate surface area from measurements of particle number and mass concentration using readily available direct-reading instruments. By assuming a lognormal aerosol size distribution with a specific geometric standard deviation, number and mass concentration measurements may be used to estimate the surface area concentration associated with the distribution. Simulations have shown that surface area estimates made on unimodal lognormal aerosols will frequently lie within 100% of the actual value. Simulations using bimodal distributions indicate estimates of surface area vary from the actual value by less than an order of magnitude. Calculations based on experimental unimodal and bimodal data confirm these findings, with estimated surface area rarely being a factor of 4 greater than the actual value, and frequently being much closer than this. These findings indicate that estimating aerosol surface area exposure using readily available number and mass concentration direct-reading instruments may be suitable for providing initial data on the magnitude of surface area exposures with minimal additional effort. This would allow the accumulation of valuable exposure-response data prior to the development and implementation of more sophisticated instrumentation to more accurately estimate surface area exposure. PMID- 12581998 TI - Factors affecting the extent of dermal absorption of solvent vapours: a human volunteer study. AB - OBJECTIVES: We have previously reported that solvent vapours can be absorbed through the skin and that the extent varies markedly and depends on the chemical. For some chemicals, the extent of absorption is significant, e.g. for 1-methoxy-2 propanol dermal absorption accounts for up to 14% of the total absorbed dose after 8 h exposure at the OES. We have conducted a second study using 2 butoxyethanol to investigate the influence of temperature, humidity and clothing on the dermal absorption of vapours. As for the first study, the extent of dermal absorption was determined by biological monitoring to measure the resultant body burden of the chemical. METHODS: Four volunteers were exposed on nine occasions. For eight of these exposures they wore air-fed half-masks to supply clean air for the inhalation route. The 'baseline' conditions (one 'whole body' and one 'skin only' exposure) were 25 degrees C, 40% relative humidity with volunteers wearing shorts and T-shirt. For each subsequent exposure, a single parameter was changed: humidity (60%, 65%), temperature (20 degrees C, 30 degrees C) or clothing (minimal and overalls). Finally, a 'industrial scenario' was conducted where volunteers wore overalls over their shorts and T-shirts and environmental conditions reflected high temperature and high humidity (30 degrees C, 60%), such as might be encountered in a tank-cleaning operation or similar. RESULTS: Results show that 'baseline' dermal absorption of 2-butoxyethanol vapour was, on average, 11% of the total absorbed dose. Higher temperature (30 degrees C, mean 14%, P = 0.03) and greater humidity (65% RH, mean 13%, P = 0.1) increased dermal absorption. The wearing of whole-body overalls did not attenuate absorption (mean 10%). By combining several factors together in the 'industrial scenario', dermal absorption of vapours was significantly increased with a mean of 39% of the total absorbed dose. CONCLUSIONS: The work has shown that dermal absorption of vapours can be significant and that environmental conditions may affect the absorption. Some types of protective clothing may not be suitable to reduce absorption. The possibility of dermal absorption of vapours should be considered particularly for workers in high vapour concentration conditions where control of exposure relies on respiratory protection. PMID- 12581999 TI - On the inertial range of particles under the influence of local exhaust hoods. AB - This paper presents results from numerical simulations conducted to estimate the inertial range (stopping distance) of large aerosol particles ejected away from local exhaust hoods. Potential flow theory is used to specify the air velocity along the centerline of a flanged circular hood. A high-order Gear method is used to integrate the particle equations of motion with drag coefficients in the transitional range. The results allow for a relative comparison of hood performance based on the energy cost needed to stop the particle within a specified distance. Hood size is shown to be a critical factor, while the capture velocity generated by the hood at the point of particle ejection is of secondary importance. PMID- 12582000 TI - Partitioning theory for respiratory deposition of semivolatile aerosols. AB - The objective of this work is to model the deposition of semivolatile aerosols in the lungs based on gas-particle and tissue-air partitioning theory. Semivolatile compounds exist in air as both particles and gases simultaneously. Mass distributes between the two phases according to a gas-particle partitioning ratio, R(pg) = K(p)(TSP). Particle deposition in the lungs is a function of aerodynamic diameter, whereas gas deposition is a function of tissue solubility, which is related to the air-lung partitioning ratio, K(la). Therefore, deposition to the lungs will vary with R(pg) and K(la). These and other parameters determine a dimensionless deposition number, D, which indicates whether particles or gases are most responsible for deposition of semivolatile chemicals in the lung. The deposition number allows industrial hygienists to design effective air sampling strategies and control measures that will minimize risks associated with exposure to semivolatiles. Examples of deposition numbers for common semivolatile pollutants are provided, including alkanes, polycyclic aromatic hydrocarbons, pesticides and polychlorinated biphenyls. PMID- 12582001 TI - Comment on Sartorelli et Al. (1998): invalid calculation of permeability coefficients. PMID- 12582004 TI - Sodium loading changes urinary protein excretion: a proteomic analysis. AB - Plasma sodium concentration is maintained even when sodium intake is altered. Sodium homeostasis may involve changes in renal tubular protein expression that are reflected in the urine. We used proteomic analysis to investigate changes in urinary protein excretion in response to acute sodium loading. Rats were given deionized water followed by hypertonic (2.7%) saline for 28 h each. Urinary protein expression was determined during the final 4 h of each treatment. Acute sodium loading increased urinary sodium excretion (4.53 +/- 1.74 vs. 1.70 +/- 0.27 mmol/day, P = 0.029). Urinary proteins were separated by two-dimensional PAGE and visualized by Sypro ruby staining. Differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry followed by peptide mass fingerprinting. The abundance of a total of 45 protein components was changed after acute sodium loading. Neutral endopeptidase, solute carrier family 3, meprin 1alpha, diphor-1, chaperone heat shock protein 72, vacuolar H(+)-ATPase, ezrin, ezrin/radixin/moesin-binding protein, glutamine synthetase, guanine nucleotide-binding protein, Rho GDI-1, and chloride intracellular channel protein 1 were decreased, whereas albumin and alpha-2u globulin were increased. Some of these proteins have previously been shown to be associated with tubular transport. These data indicate that alterations in the excretion of several urinary proteins occur during acute sodium loading. PMID- 12582006 TI - Role of MAPK pathways in light chain-induced cytokine production in human proximal tubule cells. AB - We previously demonstrated that light chain (LC) endocytosis by human proximal tubule cells (PTCs) leads to production of cytokines through activation of NF kappaB. Here, we examined the role of MAPK pathways in these responses using four species of myeloma LCs (kappa(1), kappa(2), kappa(3), and lambda(1)) previously shown to induce cytokine production by PTCs. Among these, kappa(1)-LC, which yielded the strongest cytokine responses, was selected for detailed studies. Activation of MAPKs was probed by Western blot analysis for the active kinases, ERK 1/2, JNK 1/2, and p38 in kappa(1)-LC-exposed human PTCs. To evaluate the functional role of MAPKs in LC-induced cytokine responses, we tested the effects of U-0126, an ERK inhibitor; SP-600125, an inhibitor of JNK; SB-203580, a p38 inhibitor; and curcumin, a JNK-AP-1 inhibitor, all added to media before 4-h exposure to 1.5 mg/ml kappa(1)-LC. IL-6 and monocyte chemotactic protein-1 (MCP 1) were determined by ELISA. Both LC and human serum albumin (HSA) activated ERK, although the HSA effect was weaker. kappa(1)-LC stimulated all three MAPKs, although phosphorylation of ERK was more pronounced and sustained than others. Inhibitors of ERK, JNK, and p38 reduced LC-induced IL-6 and MCP-1 production. These findings suggest that activation of MAPKs plays a role in LC-induced cytokine responses in PTCs. Activation of MAPKs may be involved in cytokine responses induced by other proteins as well as LCs and may be pivotal in the pathophysiology of tubulointerstitial injury in proteinuric diseases. PMID- 12582005 TI - NO inhibits Na+-K+-2Cl- cotransport via a cytochrome P-450-dependent pathway in renal epithelial cells (MMDD1). AB - Nitric oxide (NO) exerts direct effects on nephron transport. We determined the effect of NO on Na(+)-K(+)-2Cl(-) cotransport in a cell line (MMDD1) with properties of macula densa. Na(+)-K(+)-2Cl(-) cotransport was measured as bumetanide-sensitive (86)Rb(+) uptake in the presence of ouabain. MMDD1 cells expressed mRNA for the neuronal isoform of nitric oxide synthase, as well as NKCC1 and NKCC2(B) isoforms of the Na(+)-K(+)-2Cl(-) cotransporter. Preincubation of cells with the NO donors sodium nitroprusside (SNP) or S-nitroso-N acetylpenicillamine (SNAP) caused concentration-dependent inhibition of Na(+) K(+)-2Cl(-) cotransport. Both apical and basolateral Na(+)-K(+)-2Cl(-) cotransport was inhibited by NO donors. SNP or SNAP had no significant effect on cellular levels of cGMP, cAMP, cytosolic calcium, or phosphorylation of ERK1 and ERK2. In contrast, the inhibitors of cytochrome P-450, 1-aminobenzotriazole (ABT; 10(-3) M) or ketoconazole (1.5 x 10(-5) M), completely reversed the inhibitory effect of SNAP on apical or basolateral Na(+)-K(+)-2Cl(-) cotransport [apical: control 1.18 +/- 0.15 vs. SNAP (10(-4) M) 0.41 +/- 0.05 pmol x mg(-1) x 5 min( 1); P < 0.001; SNAP (10(-4) M) + ABT 1.32 +/- 0.10 pmol x mg(-1) x 5 min(-1); P = not significant vs. control; n = 5]. The cytochrome P-450 epoxyeicosatrienoic acid (EET) metabolite 14,15-EET (5 x 10(-7) M) inhibited both apical and basolateral cotransport, whereas 8,9-EET and 11,12-EET had no significant effect. Although 20-hydroxyeicosatetraenoic acid inhibited apical cotransport, the inhibitor of omega-hydroxylase activity HET0016 did not reverse SNAP-mediated inhibition of apical cotransport. These data indicate that NO inhibits apical and basolateral Na(+)-K(+)-2Cl(-) cotransport in MMDD1 cells. The results suggest that the inhibitory pathway is independent of cGMP and might involve stimulation of a cytochrome P-450-dependent pathway. PMID- 12582007 TI - Renal function in NHE3-deficient mice with transgenic rescue of small intestinal absorptive defect. AB - The degree to which loss of the NHE3 Na(+)/H(+) exchanger in the kidney contributes to impaired Na(+)-fluid volume homeostasis in NHE3-deficient (Nhe3(-/ )) mice is unclear because of the coexisting intestinal absorptive defect. To more accurately assess the renal effects of NHE3 ablation, we developed a mouse with transgenic expression of rat NHE3 in the intestine and crossed it with Nhe3( /-) mice. Transgenic Nhe3(-/-) (tgNhe3(-/-)) mice tolerated dietary NaCl depletion better than nontransgenic knockouts and showed no evidence of renal salt wasting. Unlike nontransgenic Nhe3(-/-) mice, tgNhe3(-/-) mice tolerated a 5% NaCl diet. When fed a 5% NaCl diet, tgNhe3(-/-) mice had lower serum aldosterone than tgNhe3(-/-) mice on a 1% NaCl diet, indicating improved extracellular fluid volume status. Na(+)-loaded tgNhe3(-/-) mice had sharply increased urinary Na(+) excretion, reflective of increased absorption of Na(+) in the small intestine; nevertheless, they remained hypotensive, and renal studies showed a reduction in glomerular filtration rate (GFR) similar to that observed in nontransgenic Nhe3(-/-) mice. These data show that reduced GFR, rather than being secondary to systemic hypovolemia, is a major renal compensatory mechanism for the loss of NHE3 and indicate that loss of NHE3 in the kidney alters the set point for Na(+)-fluid volume homeostasis. PMID- 12582008 TI - Elevated plasma nonesterified fatty acids are associated with deterioration of acute insulin response in IGT but not NGT. AB - High concentrations of nonesterified fatty acids (NEFA) are a risk factor for developing type 2 diabetes in Pima Indians. In vitro and in vivo, chronic elevation of NEFA decreases glucose-stimulated insulin secretion. We hypothesized that high fasting plasma NEFA would increase the risk of type 2 diabetes by inducing a worsening of glucose-stimulated insulin secretion in Pima Indians. To test this hypothesis, fasting plasma NEFA concentrations, body composition, insulin action (M), acute insulin response (AIR, 25-g IVGTT), and glucose tolerance (75-g OGTT) were measured in 151 Pima Indians [107 normal glucose tolerant (NGT), 44 impaired glucose tolerant (IGT)] at the initial visit. These subjects, participants in ongoing studies of the pathogenesis of obesity and type 2 diabetes, had follow-up measurements of body composition, glucose tolerance, M, and AIR. In NGT individuals, cross-sectionally, high fasting plasma NEFA concentrations at the initial visit were negatively associated with AIR after adjustment for age, sex, percent body fat, and M (P = 0.03). Longitudinally, high fasting plasma NEFA concentrations at the initial visit were not associated with change in AIR. In individuals with IGT, cross-sectionally, high fasting plasma NEFA concentrations at the initial visit were not associated with AIR. Longitudinally, high fasting plasma NEFA concentrations at the initial visit were associated with a decrease in AIR before (P < 0.0001) and after adjustment for sex, age at follow-up, time of follow-up, change in percent body fat and insulin sensitivity, and AIR at the initial visit (P = 0.0006). In conclusion, findings in people with NGT indicate that fasting plasma NEFA concentrations are not a primary etiologic factor for beta-cell failure. However, in subjects who have progressed to a state of IGT, chronically elevated NEFA seem to have a deleterious effect on insulin-secretory capacity. PMID- 12582009 TI - Human placenta metabolizes fatty acids: implications for fetal fatty acid oxidation disorders and maternal liver diseases. AB - The role of fat metabolism during human pregnancy and in placental growth and function is poorly understood. Mitochondrial fatty acid oxidation disorders in an affected fetus are associated with maternal diseases of pregnancy, including preeclampsia, acute fatty liver of pregnancy, and the hemolysis, elevated liver enzymes, and low platelets syndrome called HELLP. We have investigated the developmental expression and activity of six fatty acid beta-oxidation enzymes at various gestational-age human placentas. Placental specimens exhibited abundant expression of all six enzymes, as assessed by immunohistochemical and immunoblot analyses, with greater staining in syncytiotrophoblasts compared with other placental cell types. beta-Oxidation enzyme activities in placental tissues were higher early in gestation and lower near term. Trophoblast cells in culture oxidized tritium-labeled palmitate and myristate in substantial amounts, indicating that the human placenta utilizes fatty acids as a significant metabolic fuel. Thus human placenta derives energy from fatty acid oxidation, providing a potential explanation for the association of fetal fatty acid oxidation disorders with maternal liver diseases in pregnancy. PMID- 12582010 TI - Measuring gluconeogenesis using a low dose of 2H2O: advantage of isotope fractionation during gas chromatography. AB - The contribution of gluconeogenesis to glucose production can be measured by enriching body water with (2)H(2)O to approximately 0.5% (2)H and determining the ratio of (2)H that is bound to carbon-5 vs. carbon-2 of blood glucose. This labeling ratio can be measured using gas chromatography-mass spectrometry after the corresponding glucose carbons are converted to formaldehyde and then to hexamethylenetetramine (HMT). We present a technique for integrating ion chromatograms that allows one to use only 0.05% (2)H in body water (i.e., 10 times less than the current dose). This technique takes advantage of the difference in gas chromatographic retention times of naturally labeled HMT and [(2)H]HMT. We discuss the advantage(s) of using a low dose of (2)H(2)O to quantify the contribution of gluconeogenesis. PMID- 12582011 TI - NPY ablation in C57BL/6 mice leads to mild obesity and to an impaired refeeding response to fasting. AB - Neuropeptide Y (NPY) is an orexigenic (appetite-stimulating) peptide that plays an important role in regulating energy balance. When administered directly into the central nervous system, animals exhibit an immediate increase in feeding behavior, and repetitive injections or chronic infusions lead to obesity. Surprisingly, initial studies of Npy(-/-) mice on a mixed genetic background did not reveal deficits in energy balance, with the exception of an attenuation in obesity seen in ob/ob mice in which the NPY gene was also deleted. Here, we show that, on a C57BL/6 background, NPY ablation is associated with an increase in body weight and adiposity and a significant defect in refeeding after a fast. This impaired refeeding response in Npy(-/-) mice resulted in a deficit in weight gain in these animals after 24 h of refeeding. These data indicate that genetic background must be taken into account when the biological role of NPY is evaluated. When examined on a C57BL/6 background, NPY is important for the normal refeeding response after starvation, and its absence promotes mild obesity. PMID- 12582012 TI - Effect of heart failure on the regulation of skeletal muscle protein synthesis, breakdown, and apoptosis. AB - Heart failure is often characterized by skeletal muscle atrophy. The mechanisms underlying muscle wasting, however, are not fully understood. We studied 30 Dahl salt-sensitive rats (10 male, 20 female) fed either a high-salt (HS; n = 15) or a low-salt (LS; n = 15) diet. This strain develops cardiac hypertrophy and failure when fed a HS diet. LS controls were matched to HS rats for gender and duration of diet. Body mass, food intake, and muscle mass and composition were measured. Skeletal muscle protein synthesis was measured by isotope dilution. An additional group of 27 rats (HS, n = 16; LS; n = 11) were assessed for expression of genes regulating protein breakdown and apoptosis. Gastrocnemius and plantaris muscles weighed less (16 and 22%, respectively) in HS than in LS rats (P < 0.01). No differences in soleus or tibialis anterior weights were found. Differences in muscle mass were abolished after data were expressed relative to body size, because HS rats tended (P = 0.094) to weigh less. Lower body mass in HS rats was related to a 16% reduction (P < 0.01) in food intake. No differences in muscle protein or DNA content, the protein-to-DNA ratio, or muscle protein synthesis were found. Finally, no differences in skeletal muscle gene expression were found to suggest increased protein breakdown or apoptosis in HS rats. Our results suggest that muscle wasting in this model of heart failure is not associated with alterations in skeletal muscle metabolism. Instead, muscle atrophy was related to reduced body weight secondary to decreased food intake. These findings argue against the notion that heart failure is characterized by a skeletal muscle myopathy that predisposes to atrophy. PMID- 12582013 TI - Differential activation of NF-kappa B and AP-1 in increased fibronectin synthesis in target organs of diabetic complications. AB - Increased extracellular matrix protein production leading to structural abnormalities is a characteristic feature of chronic diabetic complications. We previously showed that high glucose in endothelial cell culture leads to the upregulation of basement membrane protein fibronectin (FN) via an endothelin (ET) dependent pathway involving activation of NF-kappaB and activating protein-1 (AP 1). To delineate the mechanisms of basement membrane thickening, we used an animal model of chronic diabetes and evaluated ET-dependent activation of NF kappaB and AP-1 and subsequent upregulation of FN in three target organs of chronic diabetic complications. After 3 mo of diabetes, retina, renal cortex, and myocardium demonstrated increased FN mRNA and increased ET-1 mRNA expression. Increased FN expression was shown to be dependent on ET receptor-mediated signaling, as the increase was prevented by the dual ET receptor antagonist bosentan. NF-kappaB activation was most pronounced in the retina, followed by kidney and heart. AP-1 activation was also most pronounced in the retina but was similar in both kidney and heart. Bosentan treatment prevented NF-kappaB activation in the retina and heart and AP-1 activation in the retina and kidney. These data indicate that, although ETs are important in increased FN production due to diabetes, the mechanisms with respect to transcription factor activation may vary depending on the microenvironment of the organ. PMID- 12582014 TI - Regulation of type II deiodinase expression by EGF and glucocorticoid in HC11 mouse mammary epithelium. AB - Thyroid hormones are important for mammary gland growth and development. The iodothyronine deiodinases play a key role in thyroid hormone metabolism. We have showed that type II 5'-deiodinase (5'D2) activity and mRNA are present in the mouse mammary gland and that their levels are reduced in the lactating gland. To investigate the regulatory mechanism of mouse 5'D2 gene (mdio2) expression in mammary epithelium, we employed the HC11 cell line, which is derived from mouse mammary epithelial cells and retains the ability to express differentiated function. HC11 cells were treated with combinations of insulin, glucocorticoid (GC, dexamethasone), prolactin, and epidermal growth factor (EGF), and 5'D2 activity and the D2-to-GAPDH mRNA ratio were measured by (125)I(-) release from (125)I-labeled thyroxine and semiquantitative RT-PCR, respectively. EGF increased both 5'D2 activity and mRNA levels about twofold. GC reduced both 5'D2 activity and mRNA in a dose-dependent manner, and their levels were decreased to approximately one-tenth and one-fifth, respectively, of control levels. These data demonstrated that mdio2 expression in HC11 cells is upregulated by EGF mainly at the pretranslational level and downregulated by GC at both pre- and posttranslational levels. Furthermore, we showed that GC reduced the promoter activity of the 627- bp 5'-upstream region of the mdio2/luciferase chimeric reporter gene, suggesting that GC exerts its effect, at least in part, at the transcriptional level. PMID- 12582015 TI - Gastrointestinal cancer prevention in the United States: the road ahead. PMID- 12582016 TI - Nutritional predictors of insulin-like growth factor I and their relationships to cancer in men. AB - The insulin-like growth factor (IGF) axis may play opposing roles in health and disease. The age-related declines in growth hormone and IGF-I may be associated with potentially deleterious changes in body composition and functioning, but recent studies suggest that IGF-I levels may be related to risk of prostate, colorectal, premenopausal breast, and possibly other cancers. Thus, we studied dietary influences on plasma IGF-I and IGF-I:IGF-binding protein-3 ratio in 753 men in the Health Professionals Follow-Up Study who completed a food frequency questionnaire. In this generally well-nourished population of middle-aged to elderly men, plasma IGF-I and IGF-I:IGF-binding protein-3 molar ratio tended to increase with higher intake of protein and minerals, including potassium, zinc, magnesium, calcium, and phosphorus. Men with relatively high intakes of total protein (top quintile) and minerals (top quintile of the five minerals combined) had a 25% higher mean plasma level of IGF-I compared with those in the low quintiles simultaneously. The major sources of animal protein, including milk, fish, and poultry, but not red meat, as well as total vegetable protein, were associated with an increase in IGF-I levels. Energy intake was positively related to plasma IGF-I level but only in men with body mass index <25 kg/m(2). The age related decline in plasma IGF-I may be exacerbated by low intakes of protein and minerals. The potential role of these dietary factors on cancer risk through altering IGF-I levels requires study. PMID- 12582018 TI - Intention to pursue genetic testing for breast cancer among women due for screening mammography. AB - Because few studies have addressed the intention to pursue testing for breast cancer susceptibility among women in the general population, we examined whether women due for routine mammography would want such testing and what factors might impact on their decision to pursue testing. A questionnaire was mailed to women > or =50 years of age who had undergone a screening mammogram 12 to 14 months before the study. Univariate and multivariable analyses were conducted to identify factors associated with intention to pursue genetic testing. Approximately 41% of respondents probably or definitely intended to pursue testing. In univariate analysis, the intention to undergo testing was not significantly associated with age, education, marital status, potential effects on health or life insurance, or physician recommendation. Although significant in univariate analysis, family history of breast cancer and ethnicity were not significant in multivariable analysis. In both univariate and multivariable analysis, factors significantly associated with intention to undergo testing included awareness of genetic testing, cancer worry, and insurance coverage of testing cost. Intention also was associated with the respondent wanting to know whether she possessed the susceptibility gene, even if that knowledge would not impact on options for early detection or treatment. Given the relatively high level of interest in testing among women at average risk of breast cancer, these results may help health care professionals educate and counsel women regarding the appropriate use of genetic testing as well as breast cancer risk factors. PMID- 12582019 TI - Soluble epidermal growth factor receptor (sEGFR/sErbB1) as a potential risk, screening, and diagnostic serum biomarker of epithelial ovarian cancer. AB - Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancies in the United States, for which risk assessment, screening, and diagnostic tests are needed. We have shown previously that women with stage III/IV EOC have lower serum p110 sEGFR/sErbB1 (Soluble Epidermal Growth Factor Receptor) concentrations than healthy women. Here, we show that serum p110 sEGFR/sErbB1 is the product of a 3-kb EGFR/ERBB1 alternate transcript. We report that serum sEGFR concentrations in stage I/II and stage III/IV EOC patients are significantly lower than in healthy women, and that serum sEGFR concentrations are not associated with disease stage or tumor grade. Logistic regression models show that: (a) lower serum sEGFR concentrations are associated significantly with a greater risk of EOC; (b) the risk associated with lower serum sEGFR concentrations is reduced by older age or menopause; and (c) age- or menopausal status-specific cutoff values for sEGFR concentration are appropriate. Receiver operating characteristic curves indicate that: (a) serum sEGFR concentrations are more effective in discerning stage III/IV than stage I/II EOC cases from healthy women; and (b) sEGFR concentrations have an 89% probability of correctly discerning EOC patients from healthy women when accounting for effect modification by age. By maintaining a test specificity of approximately 95% across strata of age or menopausal status with appropriate cutoff values, we observe that sEGFR concentrations are most useful for detecting stage I/II (sensitivity: 64-67%) and stage III/IV (sensitivity: 75-81%) EOC in young, premenopausal women. We conclude that serum sEGFR concentrations warrant additional investigation in the risk assessment, early detection, and/or diagnosis of EOC. PMID- 12582017 TI - BRCA1 wild-type allele modifies risk of ovarian cancer in carriers of BRCA1 germ line mutations. AB - Strong inter- and intrafamilial variation of penetrance of breast and ovarian cancer is observed in BRCA1 mutation carriers. The wild-type copy of the BRCA1 gene is a plausible candidate as a cancer risk modifier given that the residual function corresponding to the intact BRCA1 allele may influence the process of tumor formation in BRCA1 carriers. Indeed, growing evidence is now becoming available on impaired reparation of double-strand DNA breaks in cells heterozygous for BRCA1 mutations, implying an enhanced mutability of BRCA1(+/-) cells. To determine whether certain variant forms of the wild-type BRCA1 allele are implicated in variation of the BRCA1-related cancer risk, their effect was studied in a panel of 591 women with BRCA1 germ-line mutations. We found that BRCA1 carriers with the wild-type BRCA1 copy bearing a common Gly1038 variant were at increased risk of ovarian cancer (hazards ratio, 1.50; 95% confidence interval, 1.03-2.19). The results of our study imply that a quite significant proportion of the interindividual variability in ovarian cancer penetrance in BRCA1 carriers may be explained by a common BRCA1 Gly1038 wild-type allele, given its high frequency (0.27). PMID- 12582020 TI - The efficacy of 9-cis-retinoic acid (aliretinoin) as a chemopreventive agent for cervical dysplasia: results of a randomized double-blind clinical trial. AB - 9-Cis-retinoic acid (aliretinoin) is a pan-retinoid receptor agonist and has been demonstrated in preclinical models to have potent chemoprevention effects. The purpose of this study was to determine the utility of using aliretinoin as a chemoprevention agent in cervical dysplasia. Patients with histological evidence of cervical intraepithelial neoplasia (CIN) 2/3 were randomized in a double-blind manner to receive high-dose aliretinoin (50 mg), low-dose of aliretinoin (25 mg), or placebo daily for 12 weeks. Compliance and side effects were monitored at various time points during therapy. At the completion of therapy, all of the patients underwent a loop procedure. Histology of pretreatment biopsies was compared with that of loop specimens. One-hundred and fourteen patients with CIN 2/3 were enrolled in the study. In the 112 patients evaluable for toxicity, headache was the most common clinical side effect and was experienced more frequently (74%) in the high-dose aliretinoin group. Eight patients withdrew from the study before completion of study medication because of unacceptable side effects. In the 104 patients evaluable for efficacy, there was no statistical difference in the rate of regression among the placebo (32%), the low-dose aliretinoin (32%), and the high-dose aliretinoin (36%) groups. (P = not significant; power 0.06). Aliretinoin at these dosages and this schedule does not appear to result in significant regression rates in CIN 2/3 patients when compared with placebo. Headache is encountered frequently and may thwart efforts to increase the dose or duration of aliretinoin in future cervical cancer chemoprevention studies. The rate of histological regression in biopsied CIN 2/3 patients is high even over a short time interval, and emphasizes the importance of having a placebo arm and an adequate sample size in cervical dysplasia chemoprevention studies. PMID- 12582021 TI - Association of the CYP17 gene polymorphism with the risk of prostate cancer: a meta-analysis. AB - A T-to-C polymorphism in the 5' promoter region of the CYP17 gene that encodes the cytochrome P450c17alpha has been implicated as a risk factor for prostate cancer, but individual studies have been inconclusive or controversial. Therefore we performed a meta-analysis of 10 studies (12 comparisons) with CYP17 genotyping on 2404 patients with prostate cancer and 2755 controls. Overall, the random effects odds ratio (OR) for the A2 (C) versus A1 (T) allele was 1.08 [95% confidence interval (CI), 0.95-1.22], with some between-study heterogeneity (P = 0.04). There was no suggestion of an overall effect either in recessive or dominant modeling of A2 effects, and the comparison of A2/A2 versus A1/A1 also showed no differential susceptibility to prostate cancer (OR, 1.15; 95% CI, 0.91 1.46). No effect of A2 was seen in subjects of European descent (7 comparisons, OR, 1.04; 95% CI, 0.92-1.18, no significant between-study heterogeneity) or Asian descent (2 comparisons, OR, 1.06; 95% CI, 0.66-1.71; P = 0.02 for heterogeneity), whereas A2 increased susceptibility to prostate cancer in subjects of African descent (3 comparisons, OR, 1.56; 95% CI, 1.07-2.28; no between-study heterogeneity). Smaller studies unilaterally showed more prominent genetic effects for A2 than larger studies (P = 0.038). The meta-analysis suggests that the CYP17 polymorphism is unlikely to increase considerably the risk of sporadic prostate cancer on a wide population basis, especially in subjects of European descent. Previously reported associations may reflect publication bias, although it is also possible that the polymorphism may be important in subjects of African descent. PMID- 12582022 TI - Androgen receptor (CAG)n and (GGC)n polymorphisms and breast cancer risk in a population-based case-control study of young women. AB - The androgen receptor (AR) is involved in the regulation of hormone-responsive genes and, as such, variation within the gene is hypothesized to play a role in breast cancer susceptibility. We therefore assessed the relationship between AR repeat variation and breast cancer in young women from the general population. Women diagnosed with breast cancer before age 45 years and age-matched controls, all participants in a population-based case-control study of breast cancer, were assessed for length variation in the (CAG)(n) and (GGC)(n) AR repeats within the AR gene. Results were generated from 524 cases and 461 controls. As per previous studies, (CAG)(n) repeat lengths of <22 were classified as short (S), and those of > or =22 were classified as long (L). For (GGC)(n) repeats, those < 17 were classified as short, and those > or = 17 were classified as long. Women with a cumulative (CAG)(n) repeat size of > or =43 showed a modest increase in risk for breast cancer [odds ratio (OR), 1.3; 95% confidence interval (CI), 1.0-1.7]. Women with a (GGC)(n) long (L) allele and those with a > or =33 cumulative repeat size had a decreased risk of breast cancer (OR, 0.7; 95% CI, 0.5-0.9). Among women homozygous for the (CAG)(n) short (S) allele and those with any (GGC)(n) L allele, an increased risk of breast cancer in relation to ever use of oral contraceptives [OCs; OR = 1.9 (95% CI, 1.0-3.6) and OR = 1.7 (95% CI, 0.9-3.5), respectively] was observed. An increased risk for OC use, however, was not observed among women with the CAG L or GGC S allele. This study, one of the first to examine both (CAG)(n) and (GGC)(n) in a population-based study for its relation to breast cancer risk, suggests a reduced risk in young women with (GGC)(n) repeat lengths of > or =17. In addition, these data suggest that AR repeat length may be partly responsible for the increased risk for early-onset breast cancer in women who use OCs, although these findings need replication in other populations. PMID- 12582023 TI - Enhancement of antibody detection in cancer using panel of recombinant tumor associated antigens. AB - Cancer sera contain antibodies which react with a unique group of autologous cellular antigens called tumor-associated antigens (TAAs). This study determines whether a mini-array of multiple TAAs would enhance antibody detection and be a useful approach to cancer detection and diagnosis. The mini-array of TAAs comprised full-length recombinant proteins expressed from cDNAs encoding c-myc, p53, cyclin B1, p62, Koc, IMP1, and survivin. Enzyme immunoassay was used to detect antibodies in 527 sera from six different types of cancer. Antibody frequency to any individual TAA was variable but rarely exceeded 15-20%. With the successive addition of TAAs to a final total of seven antigens, there was a stepwise increase of positive antibody reactions up to a range of 44-68%. Breast, lung, and prostate cancer patients showed separate and distinct profiles of reactivity, suggesting that uniquely constituted antigen mini-arrays might be developed to distinguish between some types of cancer. Distinct antibody profiles were not observed in gastric, colorectal, and hepatocellular carcinomas with this set of seven TAAs. Detection of autoantibodies in cancer can be enhanced by using a mini-array of several TAAs as target antigens. Additional studies in early cancer patients and high-risk individuals and the design of unique antigen panels for different cancers would help to determine whether multiple antigen mini arrays for the detection of autoantibodies might contribute a clinically useful noninvasive approach to cancer detection and diagnosis. PMID- 12582024 TI - Why are thyroid cancer rates so high in southeast asian women living in the United States? The bay area thyroid cancer study. AB - The purpose of this study is to understand why thyroid cancer incidence rates are higher among Southeast Asian (SA) women living in the United States than among other United States women. A multiethnic population-based, case-control study of thyroid cancer among women ages 20-74 was conducted in the San Francisco Bay Area. Cases diagnosed between 1992 and 1998 were identified through the area's population-based cancer registry. Controls were identified using random digit dialing and matched to cases on age and ethnicity. Asian women were classified as SA (n = 214) or Northern Asian (n = 196) based on self-reported ethnicity. Relative attributable risks, by age group (<50 and 50+), were calculated to assess what proportion of the difference in incidence rates between these populations could be attributed to the prevalence of specific thyroid cancer risk factors, assuming common relative risks across ethnic groups. Among younger women, a history of goiter or thyroid nodules and lower consumption of isoflavones from soy-based foods account for 66% of the difference in incidence between SA and Northern Asian women. Among older women, these factors, along with recent migration, accounted for 95% of the difference between these groups. When comparing SA with Caucasian women, goiter/nodules and lower consumption of carotenoids explained 67% of the difference in incidence in younger women, whereas goiter/nodules and socioeconomic variables explained 81% of the difference in incidence in older women. A greater prevalence of goiter and thyroid nodules accounts for a substantial portion of the higher thyroid cancer incidence rates among SA women. Dietary patterns also contribute to the rate differences. PMID- 12582025 TI - Chemopreventive effects of alpha-santalol on skin tumor development in CD-1 and SENCAR mice. AB - Studies from our laboratory have indicated skin cancer chemopreventive effectsof sandalwood oil in CD-1 mice. The purpose of this investigation was to study the skin cancer chemopreventive effects of alpha-santalol, a principal component of sandalwood oil in CD-1 and SENCAR mice. alpha-Santalol was isolated from sandalwood oil by distillation under vacuum and characterized by nuclear magnetic resonance and gas chromatography-mass spectrometry. Chemopreventive effects of alpha-santalol were determined during initiation and promotion phase in female CD 1 and SENCAR mice. Carcinogenesis was initiated with 7,12 dimethylbenz(a)anthracene and promoted with 12-O-tetradecanoylphorbol-13-acetate (TPA). The effects of alpha-santalol treatment on TPA-induced epidermal ornithine decarboxylase (ODC) activity and (3)H-thymidine incorporation in epidermal DNA of CD-1 and SENCAR mice were also investigated. alpha-Santalol treatment during promotion phase delayed the papilloma development by 2 weeks in both CD-1 and SENCAR strains of mice. alpha-Santalol treatment during promotion phase significantly (P < 0.05) decreased the papilloma incidence and multiplicity when compared with control and treatment during initiation phase during 20 weeks of promotion in both CD-1 and SENCAR strains of mice. alpha-Santalol treatment resulted in a significant (P < 0.05) inhibition in TPA-induced ODC activity and incorporation of (3)H-thymidine in DNA in the epidermis of both strains of mice. alpha-Santalol significantly prevents papilloma development during promotion phase of 7,12-dimethylbenz(a)anthracene-TPA carcinogenesis protocol in both CD-1 and SENCAR mice, possibly by inhibiting TPA-induced ODC activity and DNA synthesis. alpha-Santalol could be an effective chemopreventive agent for skin cancer. Additional experimental and clinical studies are needed to investigate the chemopreventive effect of alpha-santalol in skin cancer. PMID- 12582026 TI - Bulky DNA adducts and risk of cancer: a meta-analysis. AB - We present a meta-analysis to test the hypothesis that the presence of a high level of bulky DNA adducts in tissues is associated with an increased risk of cancer in humans. Seven articles were selected that matched the selection criteria, for a total of 691 cancer patients and 632 control subjects. In five studies the cases had lung cancer, in one oral cancer, and in one bladder cancer. Six studies measured adducts in WBCs and one in normal lung tissue around tumor tissue. Six were case-control investigations, and one was a case-control study on lung cancer nested within a cohort. Current smokers showed a statistically significant difference between cases and controls, with cases having 83% higher levels of adducts than controls (95% confidence interval, 0.44-1.22). Results were negative or contradictory in ex-smokers and nonsmokers. This observation was confirmed by sensitivity analyses. Publication bias does not seem to be a problem. Despite some methodological limitations, our meta-analysis shows that current smokers with high levels of adducts have an increased risk of lung and bladder cancers. This conclusion also suggests that similar (aromatic) compounds may be involved in the etiology of both types of cancer. PMID- 12582027 TI - Alcohol, folate, methionine, and risk of incident breast cancer in the American Cancer Society Cancer Prevention Study II Nutrition Cohort. AB - Recent studies suggest that the increased risk of breast cancer associated with alcohol consumption may be reduced by adequate folate intake. We examined this question among 66,561 postmenopausal women in the American Cancer Society Cancer Prevention Study II Nutrition Cohort. A total of 1,303 incident cases had accrued during the first 5 years of follow-up. Cox proportional hazards models and stratified analysis were used to examine the relationship between alcohol, dietary and total folate intake, multivitamin use, dietary methionine, and breast cancer. We observed an increasing risk of breast cancer with increasing alcohol consumption (P for trend = 0.01). In the highest category of consumption (15 or more grams of ethanol/day), the risk of breast cancer was 1.26 (95% confidence interval, 1.04-1.53) compared with nonusers. We observed this association with higher alcohol consumption for in situ, localized, and regional disease. We found no association between risk of breast cancer and dietary folate, total folate, multivitamin use, or methionine intake. Furthermore, we found no evidence of an interaction between levels of dietary folate (P for interaction = 0.10) or total folate (P for interaction = 0.61) and alcohol. Nor did we find evidence of an interaction between alcohol consumption and recent or long-term multivitamin use (P for interaction = 0.27). Our results are consistent with a positive association with alcohol but do not support an association with folate or methionine intake or an interaction between folate and alcohol intake on risk of breast cancer. PMID- 12582028 TI - The tumor suppressor gene 14-3-3 sigma is commonly methylated in normal and malignant lymphoid cells. AB - 14-3-3 sigma/Stratifin was first identified as an epithelial cell antigen (HME-1) exclusively expressed in epithelia. However, the functional role of sigma in cell proliferation and apoptosis would suggest that this protein could be relevant to the regulation of growth and differentiation of multiple cell types. Recent evidence demonstrates that sigma acts as a tumor suppressor gene that is inactivated by methylation of its 5' CpG islands in epithelial tumor cells. In normal epithelia, sigma is commonly unmethylated. The objective of this study was to determine the methylation status of sigma in lymphoid cells. We now demonstrate by methylation-specific PCR analysis that sigma is also methylated in normal and malignant lymphocytes. Such methylation, however, fails to completely silence its expression. Compared with the robust expression in epithelial cells, lymphocytes showed basal, but clearly evident, levels of sigma as determined by reverse transcription-PCR and Western blot. The finding of sigma 5' region methylation in lymphocytes has direct implications in the use of body fluids on methylation tests for noninvasive monitoring of occult epithelial tumor cells and suggests that sigma may not be an adequate biomarker for methylation-specific PCR analysis. PMID- 12582029 TI - No association between OGG1 Ser326Cys polymorphism and breast cancer risk. PMID- 12582030 TI - Association between the Met326Ile polymorphism of the p85alpha regulatory subunit of phosphatidylinositol 3-kinase and prostate cancer risk: a prospective study. PMID- 12582031 TI - No association between the p21 codon 31 serine-arginine polymorphism and lung cancer risk. PMID- 12582032 TI - Lack of association between CCND1 G870A polymorphism and risk of esophageal squamous cell carcinoma. PMID- 12582033 TI - Polio vaccination and risk of brain tumors in adults: no apparent association. PMID- 12582034 TI - No association between genetic polymorphisms of CYP1A1, GSTM1, GSTT1, GSTP1, NAT2, and nasopharyngeal carcinoma in Taiwan. PMID- 12582036 TI - Recent Developments in Cerebellar Research. November 15-18, 2001. St. Louis, Missouri, USA. Conference proceedings. PMID- 12582037 TI - Disorders in timing and force of finger opening in overarm throws made by cerebellar subjects. AB - Although there is agreement that an important sign of cerebellar dysfunction is disorder in timing of movement, it appears that authors who study different behaviors mean different things when they use the term "timing," and that the underlying mechanisms are likely to be different. For overarm throwing, skilled throwers can time ball release with a precision of less than 7 ms, whereas cerebellar subjects show a large variability of 50 ms or more in this timing. Furthermore, cerebellar patients show a larger variability in the amplitude of finger opening which could either reflect a disorder in force, or result indirectly from the increased variability in timing. To determine whether timing and force of finger opening were dependent variables, the time of ball release was plotted against the amplitude of finger opening. In control subjects these two parameters were related, with early (mistimed) throws having smaller finger amplitudes. However, in cerebellar subjects the increased variability in finger amplitude could not be accounted for by the increased variability in timing. Similarly, the increased timing windows could not be explained by disorder in force at the fingers. It is concluded that the abnormal finger opening that occurs in cerebellar patients when making overarm throws results from increased variability in both the timing and force of finger extension. Whether the increased variability in timing is a disorder in triggering finger opening at the right moment, or is due to a failure to combine finger opening appropriately with the hand trajectory, remains to be determined. PMID- 12582038 TI - Cerebellar limb ataxia: abnormal control of self-generated and external forces. AB - Our work has been focused on understanding the mechanism of movement abnormalities associated with cerebellar ataxia. The hypothesis tested is that the cerebellum acts to modulate muscle activity across multiple joints in anticipation of the mechanical interaction torques generated by one's own movement and by external forces. Individuals with cerebellar damage were studied in two sets of experiments. In the first experiment, we studied how cerebellar subjects' movement changed when interaction torques were present, and then reduced via mechanically constraining movement to a single joint. Consistent with the hypothesis, it was found that cerebellar endpoint errors were greatly improved when interaction torques were reduced. We also found that cerebellar deficits in the unconstrained condition were not explained by a general failure of torque timing or magnitude scaling. This supports the idea that the cerebellum plays a specific and important role in adjusting for the dynamics of one's own body movements. In the second experiment, we studied how well cerebellar subjects could adapt arm movements to external loads. Cerebellar subjects were tested as they adapted a catching movement to balls of different weight. It was found that they were slow or unable to adapt through practice and did not show evidence of storage of the adaptation. This suggests that the cerebellum is needed for rapid adaptations for loads in movement. Given these findings, we think that the cerebellum is important in anticipating and adjusting for the mechanical demands of movement though trial-and-error practice. PMID- 12582041 TI - The role of the oculomotor vermis in the control of saccadic eye movements. AB - The oculomotor vermis is a part of the posterior cerebellum, characterized by a low threshold (<10 micro A) for evoked saccades. It comprises vermal lobuli VIc and VIIA. Many Purkinje cells in this area show eye position or saccade-related responses or combinations of the two and usually lack responses to the presentation of visual targets, guiding the oculomotor behavior. The saccade related responses are directionally selective and show preferences for saccade amplitude or duration, which differ widely between cells. However, at the population level, these saccade-related Purkinje cells give a very precise account of the timing of the saccadic eye movement and, specifically, of the time it ends. This population signal might therefore contribute to determining the end of the saccadic eye movement. Furthermore, by changing the duration of the population response, the amplitude of the saccade could be changed. In other words, saccadic adaptation could be a consequence of changing a representation of time in the cerebellum. PMID- 12582039 TI - The nodulus and uvula: source of cerebellar control of spatial orientation of the angular vestibulo-ocular reflex. AB - The nodulus and rostral-ventral uvula of the vestibulo-cerebellum play a critical role in orienting eye velocity of the slow component of the angular vestibulo ocular reflex (aVOR) to gravito-inertial acceleration (GIA). This is done by altering the time constants of "velocity storage" in the vestibular system and by generating "cross-coupled" eye velocities that shift the eye velocity vector from along the body yaw axis to the yaw axis in a spatial frame. In this report, we show that eye velocity generated through the aVOR by constant velocity centrifugation in the monkey orients to the GIA in space, regardless of the position of the head with respect to the axis of rotation. We also show that, after removal of the nodulus and rostral-ventral uvula, the spatial orientation of eye velocity to the GIA is lost and that eye velocity is then purely driven by the semicircular canals in a body frame of reference. These findings are further confirmation that these regions of the vestibulo-cerebellum control spatial orientation of the aVOR. PMID- 12582042 TI - Role of the fastigial nucleus in controlling horizontal saccades during adaptation. AB - Saccade size was adapted in rhesus monkeys using surgical weakening of the muscles of one eye combined with monocular viewing. Neurons in the caudal fastigial nucleus were recorded during the adaptation. Neuronal discharges changed in a way that could be interpreted as causing the changes in saccade size given our current knowledge of the projections of these neurons to the saccadic burst generator in the brain stem. PMID- 12582043 TI - Cerebellar mechanisms in eyeblink conditioning. AB - A recent model of cerebellar learning in eyeblink conditioning predicts two sites of plasticity, the cerebellar cortex and cerebellar nuclei, which store information relating to timing and driving the movement, respectively. Consistent with this idea, lesions of the cortex or reversible "disconnections" of Purkinje cell output to the nuclei have been shown to disrupt response timing to produce short-latency conditioned eyeblinks. To better characterize potential cortical and nuclear plasticities, we analyzed the effects upon nictitating membrane (NM) and eyeblink conditioned responses (CRs) of different drugs administered to the cortex and to the nuclei. When either excitatory or inhibitory inputs to the cerebellar cortical lobule HVI were blocked by infusions of the AMPA receptor antagonist CNQX or the GABA-A receptor antagonists picrotoxin or SR95531, CRs were abolished. Similarly GABA-A receptor antagonists in the cerebellar nuclei abolished CRs. CR latencies were never shortened. However, blockade of AMPA/kainate receptor-mediated excitatory transmission to the nuclei had no effect upon CR frequencies or latencies. These results suggest that normal cortical and nuclear function is required for performance of NM and eyeblink CRs. We saw no evidence that CRs can be driven by AMPA/kainate receptor-mediated transmission from mossy fiber afferents to the cerebellar nuclei. So, although plasticity in the cerebellar nuclei is not ruled out, it is unlikely that a long term change in AMPA receptor-mediated transmission from mossy fiber inputs to the nuclei is an essential mechanism in eyeblink conditioning. Our findings indicate that a fully functional olivo-cortico-nuclear loop is required to express all characteristics of associatively conditioned responses. PMID- 12582044 TI - Role of the nuclei in eyeblink conditioning. AB - Evidence to date supports the strong conclusion that the cerebellum learns. Classical conditioning of the eyeblink response is critically dependent upon the cerebellum. The issue addressed here is whether cerebellar cortex or deep nuclei form the basic association. Learning occurs with large cerebellar cortical aspirations in rabbits and with a Purkinje-cell-deficient mutation in mice. The learned response is poorly timed, small in amplitude, and inconsistent in its occurrence. Learning nevertheless occurs. Lesions of the interpositus, on the other hand, prevent new learning and abolish previously learned conditioned responses. Small electrolytic lesions, kainic acid lesions, and temporary inactivation (cooling, muscimol, anisomycin) localize learning to the dorsolateral anterior interpositus nucleus. Learning-related unit activity-the signature of the engram-recorded throughout the brain depends on the interpositus. Electrical stimulation of interpositus afferents are needed for conditioning, and the conditioned interpositus has a lowered threshold. Finally, a recent anatomical study with electron microscopy shows synaptic changes in the excitatory inputs to the interpositus with conditioning. The interpositus is responsible for making the basic association between conditioned and unconditioned stimuli, which in turn allows ancillary learning to occur in cerebellar cortex, and possibly brainstem and forebrain. PMID- 12582045 TI - An experimental study of posterior interpositus involvement in the genesis and control of conditioned eyelid responses. AB - The participation of the rostral-dorsal part of the posterior interpositus nucleus in the genesis of classically-conditioned eyelid responses in alert cats was studied. Cats were prepared for the recording of left eyelid movements with the search-coil-in-a-magnetic-field technique and of the electromyographic activity of the orbicularis oculi muscle. A stimulating electrode was implanted in the contralateral red nucleus and a guide cannula in the ipsilateral interpositus. The cannula allowed unitary recordings, electrical microstimulation, drug injection, and push-pull perfusion of the selected interpositus site. Perfusate was used for amino acid analysis with an HPLC-EC method. Animals were conditioned with a delay (CS: tone, US: air puff) paradigm. Antidromically identified interpositus neurons fired during eyelid conditioned responses (CRs) with a variable delay to CR start. Some neurons (type A) increased and other (type B) decreased their firing during CR performance. Electrical microstimulation of posterior interpositus was able to increase CR amplitude when applied during CS presentation. Muscimol decreased CR amplitude in well-trained animals. The analysis of push-pull perfusate showed a significant increase in glutamate, glycine, and taurine, but not GABA levels, during conditioning in comparison with habituation and extinction periods. Results indicate an involvement of the rostral-dorsal part of the posterior interpositus nucleus in the genesis of eyelid conditioned responses, playing an enhancing or controlling role in CR performance, rather than being its initiator. PMID- 12582047 TI - The olivocerebellar system as a generator of temporal patterns. AB - The large number of diverse functions attributed to the cerebellum appears to be inconsistent with its simple, homogeneous and evolutionary preserved structure. A homogeneous structure that participates in a variety of functions implies that a common denominator underlies all of them. Since the concept of precise timing can be recognized in almost all cerebellar functions, it is likely, therefore, that the basic cerebellar circuit is capable of generating temporal patterns. Of the different mechanisms that can generate temporal patterns, two are suggested by the functional anatomy of the cerbellum: transmission lines or oscillators. Our recent experimental observations indicate that the olivary oscillatory property is more likely to serve this function. We propose that interactions between the cerebellum and the inferior olive endow the system with the ability to generate complex temporal patterns. These temporal patterns can be used for fine adjustment of motor output, sensory expectation, or shifting attentions. PMID- 12582048 TI - The organization of cerebellar cortical circuitry revisited: implications for function. AB - For more than 35 years there has been experimental evidence that parallel fiber activity does not generate the beams of activated Purkinje cells hypothesized on the basis of cortical anatomy and assumed by most theories of cerebellar cortical function. This paper first reviews the evidence for and against the parallel fiber beam hypothesis, and then discusses the findings of our recent experimental and model-based investigations intended to better understand parallel fiber effects on Purkinje cells. A principal conclusion of these studies is that the excitatory effects of parallel fibers on Purkinje cell dendrites are modulating and must be considered in the context of a balancing inhibitory influence provided by molecular layer interneurons to these same dendrites. It is proposed that this association of excitation and inhibition can account for the lack of beam-like effects on Purkinje cells. The paper concludes by considering the consequences of this new interpretation of cerebellar cortical circuitry for current theories of cerebellar function. PMID- 12582049 TI - Dynamic correlation of neuronal activity in rat cerebellar cortex modulated by behavior. AB - Sprague-Dawley rats (2-4 months old) were trained to perform a reaching-grasping task while their head was fixated and multielectrode recordings were performed in the ipsilateral cerebellar hemisphere. Multiunit (MU) activity was recorded with 2-3 electrodes in the Purkinje cell layer at various depths (1.5-4.5 mm) from the pial surface while the animal either performed the reaching-grasping task or was at rest. Recording sites were visually aligned along the transverse or the sagittal axis of Crus IIa. Excess correlations of MU spike activity were calculated at 10-ms time resolution using the joint peristimulus time histogram (JPSTH), to reveal the dynamics of cross-correlations corrected for nonstationarities in spike rates. Peak correlation amplitudes and areas were calculated separately for a time period preceding the movement and for a succeeding period during which the movement occurred. Correlations were compared across different paradigms: transversal versus sagittal alignment of recordings sites and behavior versus rest. No significant differences were found between transversally and sagittally aligned recording sites. Significant differences in peak correlation amplitude and/or peak area, however, were found both between premovement and movement time, as well as between both these time periods and periods while the animal was at rest. PMID- 12582050 TI - Temporal organization of activity in the cerebellar cortex: a manifesto for synchrony. AB - The issues of temporal coding and the temporal organization of activity have aroused a great deal of interest in sensory systems, cortex, thalamus, and hippocampus. Strangely, despite the important timing roles attributed to the cerebellum, little consideration has been given to the organization of activity within the cerebellar circuitry. In fact, there is evidence of a remarkable temporal patterning of activity in even the earliest cerebellar recordings. The evidence for the existence of high-frequency oscillations in the cerebellar cortex is reviewed and possible mechanisms are discussed; one involves the synchrony of parallel fiber inputs to Purkinje cells. It is shown how synchronous and oscillatory activity can enable extremely precise timing and also how they can maximize the information storage capacity of the cerebellar cortex. PMID- 12582051 TI - In defense of the cerebellum. AB - The very special intrinsic connectivity of the cerebellar cortex plays but a minor role in present-day theories of cerebellar function, and it is hardly used as a source of inspiration for experiments. It is argued here that a direct translation of structure into physiological relations inescapably leads to some propositions about cerebellar function that could be tested experimentally. PMID- 12582053 TI - Functional significance of climbing-fiber synchrony: a population coding and behavioral analysis. AB - Population coding and behavioral approaches were taken toward analyzing the functional significance of the climbing-fiber system. Analyses of neuronal interaction using the joint peristimulus time histogram showed that given a low rate of firing, the climbing-fiber system organizes itself to fire synchronously during movement--a feature that bears little or no relationship to the modulation in firing rate during movement or whether olivary neurons respond to a sensory stimulus. Moreover, the climbing-fiber system avoids synchrony during a passive sensory response but actively makes a transition into synchrony as a movement is initiated. Thus, from a functional viewpoint, the active feature of the climbing fiber system to organize into synchronously firing cell ensembles is uniquely motor. Analyses of behaving rats without an inferior olive revealed that the climbing-fiber system optimizes the timing of skilled movement by reducing reaction time and the interval between repetitive movements by 100 milliseconds. Finally, using classical delay eyeblink conditioning, it was found that the inferior olive is essential for learning about rapid sequences of events but not the same event sequence when given more slowly. It was concluded that the climbing-fiber system exerts its function through synchrony, which provides a 100 ms advantage in movement speed and the ability to learn about events that are rapidly presented in time. PMID- 12582054 TI - What do complex spikes signal about limb movements? AB - Deciphering the information or signals carried by the complex spike discharge of Purkinje cells has proven to be problematic, primarily because of low frequency discharge and lack of adequate analytical techniques. This problem is particularly acute for studies of limb movements. To this end the relationship of cerebellar Purkinje cell complex spike discharge to direction and speed were studied in a manual-tracking task. Two monkeys were trained to pursue track targets moving in one of eight directions and at one of four speeds. An analysis based on Poisson regression modeling fitted the complex spike counts during single movement trials to target direction and/or speed. Using single trial data, the Poisson modeling demonstrated that the complex spike discharge for a majority of the Purkinje cells was significantly fit to tracking direction and speed. A second analysis based on the directional distribution of position and speed errors and a Poisson regression model of complex spike discharge to tracking position and speed errors found little relationship to movement error. Comparison of the preferred direction of the complex spike discharge with that of the simple spike activity revealed a reciprocal relationship for many cells. Thus, the complex spike discharge signals both tracking direction and speed but not movement errors. Furthermore, treating complex spike counts as a Poisson process provides a powerful tool for analyzing these events in single trials, without the need for extensive averaging. PMID- 12582055 TI - Inhibitory control of olivary discharge. AB - The inferior olivary nucleus is the sole source of an entire afferent system to the cerebellum, the climbing-fiber system. Inferior olivary neurons are very sensitive to the appropriate sensory stimuli, such as light contact to the paw. Yet, when animals move about, olivary cells show little change in discharge rate. Apparently some mechanism prevents the cells from discharging to stimuli generated by the animal's own movement. The inferior olive receives a massive inhibitory input from small cells in the cerebellar and vestibular nuclei. This article reviews the results from several experiments that suggest that the inferior olive is specifically targeted by inhibitory inputs that prevent responses to stimuli resulting from self-produced movement. Oscarsson proposed that the inferior olive provides the cerebellum with information about errors of motor performance and about spinal reflexes. We argue that it is unlikely that the inferior olive provides information about movement errors, although the olive may signal the occurrence of sensory events that are likely to elicit reflex movements. Another popular theory of climbing-fiber action argues that the climbing fibers play a role in altering the strength of the parallel fiber Purkinje cell synapse. The cerebellum is important for the formation of classically conditioned responses, and input generated by the unconditioned stimulus does provide effective stimulation of olivary neurons. Although the olive does not generate the unconditioned response, it may provide the cerebellum with information necessary for the formation of conditioned responses. PMID- 12582056 TI - Complex spike activity in the flocculus signals more than the eye can see. AB - Modulation of the complex spike activity of Purkinje cells in the cerebellar flocculus can convey not only visual signals but also nonvisual signals. The nonvisual complex spike modulation, which is readily observed with vestibular stimulation of the awake rabbit in darkness, is approximately in-phase with the concomitant simple spike modulation. This nonreciprocal relationship contrasts to the reciprocal relationship found when the rabbit is afforded vision. PMID- 12582057 TI - Vestibularly evoked climbing-fiber responses modulate simple spikes in rabbit cerebellar Purkinje neurons. AB - The nodulus receives a primary vestibular afferent input from the ipsilateral labyrinth and a vestibularly related climbing-fiber input originating from the contralateral labyrinth. Previously we demonstrated that increased discharge of vestibularly evoked climbing-fiber responses (CFRs) in nodular Purkinje cells was correlated with decreased discharge of simple spikes (SSs). This left unresolved the question of whether vestibularly evoked antiphasic behavior of CFRs and SSs reflects a common neural mechanism or the activation of two separate parallel pathways. We answered this question using natural vestibular stimulation to modulate the discharge of uvula-nodular Purkinje cells recorded extracellularly in unilaterally labyrinthectomized, chloralose urethane-anesthetized rabbits. In such animals, vestibular primary afferents projecting to the uvula-nodulus as mossy fibers remained intact on the side contralateral to the unilateral labyrinthectomy. The discharge of CFRs recorded in ipsilateral nodular Purkinje cells was increased by ipsilateral roll-tilt while the discharge of SSs was increased by contralateral roll-tilt. These polarities were reversed for Purkinje cells recorded in the contralateral uvula-nodulus. The polarity of SS discharge recorded from Purkinje cells on both sides of the nodulus was opposite to that of the vestibular primary mossy-fiber afferents. SSs continued to respond to contralateral roll-tilt even when the primary vestibular afferent mossy-fiber pathway was destroyed by the unilateral labyrinthectomy. Although the discharge of SSs recorded in the contralateral uvula-nodulus was increased by contralateral roll-tilt, this modulation was reduced relative to that observed in Purkinje cells recorded in the ipsilateral uvula-nodulus. We conclude that vestibularly evoked CFRs caused the modulation of SS discharge. PMID- 12582059 TI - On the amazing olivocerebellar system. AB - Over the last four decades elegant sets of a single-cell studies, originating from various research groups, have contributed significantly to our understanding of olivary and cerebellar physiology. Nevertheless questions relating to the dynamic properties of olivocerebellar network, as a system, remain unsolved. We may be reaching the limits of what can be learned using the single-cell recordings. Further research on this subject may require study of the spatiotemporal activity profiles of ensemble neuronal activity. This paper summarizes results obtained using voltage-sensitive dye imaging in inferior olive slices, and the use of mathematical modeling to address such activity profiles. PMID- 12582060 TI - Historical review of the significance of the cerebellum and the role of Purkinje cells in motor learning. AB - Classic studies of the cerebellum before the middle of the twentieth century established the structural entity of the cerebellum and characterized its function as enabling animals and humans to carry out smooth and accurate movements, even at a high speed and without visual feedback. In the 1960s, neuronal circuit structures of the cerebellum were analyzed in detail, which promoted computational approaches toward the study of neuronal network principles of the cerebellum. In the 1970s and 1980s, vestibulo-ocular reflex adaptation, adaptive locomotion, eye blink conditioning, and learning in hand/arm movement were established as effective experimental paradigms for investigating neural mechanisms of cerebellar functions. In the 1980s, long-term depression (LTD) was discovered and considered as a memory process in the cerebellum; in the 1990s, complex signal transduction processes underlying LTD were revealed. It was also in the 1980s that computational approaches were advanced for modeling control system functions of the cerebellum. Currently, there are two alternative models proposed for VOR adaptation. In this decade, we envisage new developments toward the fusion of knowledge of the cerebellum at molecular and cellular levels and those in systems and computation. Studies of LTD will play a key role in pursuing this direction. PMID- 12582061 TI - Motor and nonmotor domains in the monkey dentate. AB - Our concepts about the organization and functions of the cerebellum have changed substantially in the last 10 years. In recent studies, we used transneuronal virus tracing techniques to demonstrate that the output of the cerebellum of primates projects via the thalamus not only to its classical motor target, the primary motor cortex, but also to "nonmotor" cortical areas in the prefrontal and posterior parietal cortex. We found that dentate neurons projecting to different cortical areas originated from localized regions of the nucleus which we termed "output channels." To compare the locations of the output channels projecting to different cortical targets, we have created an unfolded map of the dentate. This unfolded map revealed that dentate output channels were segregated into spatially separate "motor" and "nonmotor" domains. The output channels in the motor domain exclusively targeted primary motor and premotor areas of the cerebral cortex. These channels were localized in the dorsal portion of the dentate. The output channels in the nonmotor domain projected to prefrontal and posterior parietal cortical areas. The nonmotor domain was confined to the ventral portion of the dentate. In recent studies, we defined a unique molecular marker, monoclonal antibody 8B3, which appears to differentially "recognize" these two domains. Taken together, our results suggest that dentate output is organized according to the functional capabilities of its cortical targets. This organization provides the dentate nucleus with the anatomical substrate to influence not only the control of movement, but also cognitive, higher-order executive and visuospatial functions. PMID- 12582062 TI - The cerebellum and event timing. AB - Damage to the cerebellum disrupts performance on a range of tasks that require precise timing including the production of skilled movements, eyeblink conditioning, and perceptual tasks such as duration discrimination. We hypothesize that such tasks involve event timing, a form of representation in which the temporal goals are explicitly represented. For example, during finger tapping, the goal to produce evenly paced intervals invokes an explicit temporal representation of the time between successive contact points with the tapping surface. In contrast, timing can be an emergent property in other actions, reflecting temporal consistencies that arise through the control of other movement parameters. One example is continuous circle drawing, a task in which temporal consistency can be achieved by maintaining a constant angular velocity or minimizing higher-order derivatives (e.g., jerk). Temporal consistency on event and emergent timing tasks is not correlated and patients with cerebellar damage show no increase in temporal variability during continuous circle drawing. While the cerebellum likely contributes to performance of a wide range of skilled behaviors, it appears to be especially important when the tasks entail event timing. PMID- 12582063 TI - Role of the posterolateral cerebellum in language. AB - Historically, scientists have believed that the cerebellum controls only movement. However, recent evidence from neuroimaging and human lesion studies suggests that the right posterolateral cerebellar hemisphere is involved, independently of movement, in helping an individual to generate verbs for given nouns. We sought to elucidate the key factors contributing to the verb generation deficits of subjects with right posterolateral cerebellar damage and thus to better understand the specific contributions of the postero-lateral cerebellum to language. We compared the performance of subjects with focal left-sided posterolateral cerebellar lesions, those with focal right-sided posterolateral cerebellar lesions, and neurologically normal pilot control subjects on an antonym generation task, noun (category member) generation task, verb selection task, and lexical decision task. Preliminary results show that subjects with right cerebellar lesions are impaired relative to other subjects only on the antonym generation task. The results provide evidence that the right cerebellar language deficit is not due solely to deficits in "mental movement" coupled to a verb and that internal generation of a word seems to be a key factor in eliciting a deficit. In addition, a semantic processing demand may be necessary but insufficient to elicit a right cerebellar language deficit. The results support the theory that the right posterolateral cerebellar hemisphere assists the left cerebral hemisphere in helping an individual learn to generate specific types of spoken, two-word associations. The full nature of this process awaits further investigation. PMID- 12582064 TI - Role of the cerebellum in motor cognition. AB - Cerebellar data from five experiments using different groups of subjects performing the same motor learning task are presented. Positron emission tomography (PET) as well as functional magnetic resonance imaging (fMRI) was used to study changes in cerebellar activations as an effect of learning. Cerebellar brain activations obtained during the performance of a new motor task were compared to activations during the performance of the same task after as well as during practice. To account for changes in velocity and somatosensory processing as an effect of practice, two control conditions were included. Behavioral data showed that as an effect of practice performance speed as well as accuracy increased in all five experiments and groups. Neuroimaging data from adults as well as children showed differential changes in brain activations in different cerebellar areas. In all experiments an area in the left lateral cerebellum showed practice-related decreases, which were most likely related to a decrease in errors. In two experiments a highly significant correlation was found between the decrease in errors and the decrease in left cerebellar activation. An area in the right lateral cerebellum and one in the ipsilateral anterior vermis showed activations that seemed related to the level of capacity at which the subjects were performing and might refer to timing-related aspects of the task. PMID- 12582067 TI - The making of a complex spike: ionic composition and plasticity. AB - Climbing fiber (CF) activation evokes a large all-or-nothing electrical response in Purkinje cells (PCs), the complex spike. It has been suggested that the role of CFs (and thus complex spikes) is that of a "teacher" in simple learning paradigms such as associative eyeblink conditioning. An alternative hypothesis describes the olivocerebellar system as part of a timing device and denies a role of the CF input in learning. To date, neither of these hypotheses nor others can definitively be verified or discounted. Similarly, the complex spike evades a clear understanding when it comes to the cellular events underlying complex spike generation. What is known, however, is that complex spikes are associated with large dendritic calcium signals that are required for the induction of long-term depression (LTD) at the parallel fiber (PF)-PC synapse. PF-LTD is a form of long term synaptic plasticity that has been suggested to underlie certain forms of cerebellar motor learning. In contrast to the PF input, the CF input has been considered invariant. Our recent discovery of LTD at the CF input shows that complex spikes are less static than previously assumed. In addition to depression of CF-evoked excitatory postsynaptic currents, long-lasting, selective reduction of slow complex spike components could be observed after brief CF tetanization. To understand the functional implications of CF-LTD, it is crucial to know the types of currents constituting the specific complex spike components. Here we review the "anatomy" of the complex spike as well as our observations of activity dependent complex spike waveform modifications. In addition, we discuss which properties CF-LTD might add to the circuitry of the cerebellar cortex. PMID- 12582068 TI - Analysis of Cx36 knockout does not support tenet that olivary gap junctions are required for complex spike synchronization and normal motor performance. AB - Electrotonic coupling by gap junctions between neurons in the inferior olive has been claimed to underly complex spike (CS) synchrony of Purkinje cells in the cerebellar cortex and thereby to play a role in the coordination of movements. Here, we investigated the motor performance of mice that lack connexin36 (Cx36), which appears necessary for functional olivary gap junctions. Cx36 null-mutants are not ataxic, they show a normal performance on the accelerating rotorod, and they have a regular walking pattern. In addition, they show normal compensatory eye movements during sinusoidal visual and/or vestibular stimulation. To find out whether the normal motor performance in mutants reflects normal CS activity or some compensatory mechanism downstream of the cerebellar cortex, we determined the CS firing rate, climbing-fiber pause, and degree of CS synchrony. None of these parameters in the mutants differed from those in wildtype littermates. Finally, we investigated whether the role of coupling becomes apparent under challenging conditions, such as during application of the tremorgenic drug harmaline, which specifically turns olivary neurons into an oscillatory state at a high frequency. In both the mutants and wildtypes this application induced tremors of a similar duration with similar peak frequencies and amplitudes. Thus surprisingly, the present data does not support the notion that electrotonic coupling by gap junctions underlies synchronization of olivary spike activity and that these gap junctions are essential for normal motor performance. PMID- 12582069 TI - Roles of inhibitory interneurons in the cerebellar cortex. AB - The roles of inhibitory interneurons in the cerebellar cortex were investigated. First, Golgi cells were specifically eliminated in transgenic mice in which Golgi cells expressed human interleukin-2 receptor alpha subunit (IL2Ralpha). Injection of exotoxin coupled to anti-IL2Ralpha antibody in the cerebellum of the transgenic mouse eliminated Golgi cells and abolished GABA and synaptic inhibition in the granular layer. After elimination of Golgi cells, acute severe ataxia and subsequent mild motor discoordination were observed. In the latter chronic phase, NMDA receptor-mediated synaptic response was reduced in granule cells. Our findings indicate that elimination of GABAergic inhibition in the granular layer caused overexcitation of granule cells resulting in severe ataxia, and then NMDA receptors in granule cells were downregulated, compensating for the reduction of GABAergic inhibition and improving motor control. In the second part, we report on the regulation mechanism of synaptic plasticity at inhibitory synapses on Purkinje cells (PCs). Inhibitory synaptic transmission on a PC is potentiated after repetitive PC depolarization. This synaptic plasticity (rebound potentiation, RP) was suppressed when a presynaptic neuron was activated during the PC depolarization. This synaptic regulation is unique in the sense that the homosynaptic activity suppresses the induction of synaptic plasticity. The mechanism of how presynaptic activity suppresses RP was examined. GABA released from the presynaptic terminal activated not only GABA(A) receptor but also GABA(B) receptor. The latter was coupled to Gi/o proteins, which downregulated adenylyl cyclase reducing cAMP and inactivated cAMP-dependent protein kinase (PKA). Downregulation of PKA suppressed RP induction. PMID- 12582070 TI - Motor performance and motor learning in Lurcher mice. AB - In adult Lurcher mice virtually all cerebellar Purkinje cells have degenerated as a direct consequence of mutant gene action, providing a natural model for studying the effect of cerebellar cortical lesions on the generation of compensatory eye movements. Lurcher mice possess both optokinetic (OKR) and vestibular (VOR) compensatory reflexes. However, clear differences were observed in control of the OKR consisting of a large reduction in gain and a moderate increase in phase lag. Minor differences were also observed in the VOR in that gain and phase lead of the reflex were both increased in Lurcher animals. Subjecting Lurcher animals to eight days of visuovestibular training tested the assumption that increased VOR gain reflected an adaptive mechanism within remaining brainstem oculomotor pathways to compensate for the reduced OKR. Contrary to control animals, Lurcher animals were unable to modify either VOR or OKR in the course of training and therefore confirmed that an intact cerebellum is indispensable for the implementation of adaptive modifications to the oculomotor system. PMID- 12582071 TI - Trying to understand the cerebellum well enough to build one. AB - The development of an increasingly detailed computer simulation of the cerebellum is briefly described. Specific and relatively direct evaluation of the performance of this simulation is made possible by the straightforward way in which pavlovian eyelid conditioning engages the cerebellum. Inputs to the simulation are based on recordings of mossy fiber and climbing fiber responses to the stimuli used in eyelid conditioning, and the output of the simulation can be evaluated with respect to the extensively characterized behavioral properties of eyelid conditioning. Because construction of the simulation has been guided by a strong aversion to errors of commission, both failures and successes of the simulation have proven informative. The behavior of the simulation related to the inhibitory nucleo-olivary feedback connection and spontaneous activity of climbing fibers is described. A prediction of the simulation concerning extinction is confirmed by experiment. PMID- 12582072 TI - Cerebellar plasticity and the ocular following response. AB - We constructed a realistic simulation model to elucidate whether the characteristics of the cerebellar synaptic plasticity reported in vitro guide the acquisition and adaptation of the ocular following response (OFR). The model reconstructed the firing frequency of the inputs of granule cell axons (GCA), inhibitory cells (IC), and climbing fibers (CF) to cerebellar Purkinje cells for the OFR, to simulate the reported cerebellar plasticity, including long-term depression, long-term potentiation, and rebound potentiation. When the model used the same visual inputs as reported for monkeys, it successfully simulated the real characteristics of simple spikes in Purkinje cells of adult monkeys and adaptation of gain and direction. The success of our simulation relied on the temporal relationship of the synaptic weight changes when CF inputs preceded GCA and IC inputs, corresponding to the relationship reported by Chen and Thompson and reanalysis of the data of Karachot et al. The success of our simulation strongly suggests that acquisition and adaptation of the OFR arise from cerebellar plasticity. PMID- 12582073 TI - Modeling cerebellar flocculus and paraflocculus involvement in complex predictive smooth eye pursuit in monkeys. AB - The role of flocculus and paraflocculus neurons in the cerebellar control of predictive eye movements was examined using two modeling techniques. The first study characterized the dependence of individual Purkinje-cell firing patterns on oculomotor output, visual input, and response timing using multilinear regression techniques. Interestingly, no dependence on visual input was detected. Purkinje cell firing was explained by sensitivities to eye position and eye velocity alone. However, complex responses occurred when sensitivity vectors pointed in different directions. For example, some neurons showed a preference for circular pursuit in a particular rotation direction. Responses also tended to lead the eye during predictable pursuit and to lag during unpredictable, visually driven pursuit. This suggests that flocculus and paraflocculus neurons played a stronger role during predictive pursuit than visually driven pursuit. A second modeling study demonstrated how the flocculus/paraflocculus system might generate predictive pursuit. A biologically realistic neural network was simulated based on the known anatomy and physiology of this cerebellar system. It included mossy and climbing fibers with realistic responses, Purkinje cells acting on well characterized brain-stem circuits, and granule, Golgi, basket, and stellate cells with appropriate connections. The network was able to learn new pursuit trajectories based on long-term alterations in synaptic connectivity at parallel to-Purkinje synapses. Interestingly, this model was able to generate predictive pursuit without visual input based only on eye-motion input. Thus, both models provide complementary evidence for the generation of nonvisual predictive control by flocculus and paraflocculus neurons. PMID- 12582074 TI - Surface-based atlases of cerebellar cortex in the human, macaque, and mouse. AB - This study describes surface reconstructions and associated flat maps that represent the highly convoluted shape of cerebellar cortex in three species: human, macaque, and mouse. The reconstructions were based on high-resolution structural MRI data obtained from other laboratories. The surface areas determined for the fiducial reconstructions are about 600 cm(2) for the human, 60 cm(2) for the macaque, and 0.8 cm(2) for the mouse. As expected from the ribbon like pattern of cerebellar folding, the cerebellar flat maps are elongated along the axis parallel to the midline. However, the degree of elongation varies markedly across species. The macaque flat map is many times longer than its mean width, whereas the mouse flat map is only slightly elongated and the human map is intermediate in its aspect ratio. These cerebellar atlases, along with associated software for visualization and for mapping experimental data onto the atlas, are freely available to the neuroscience community (see http:/brainmap.wustl.edu). PMID- 12582075 TI - Plasticity of the vertical VOR: a system identification approach to localizing the adaptive sites. AB - The vestibulo-ocular reflex (VOR) stabilizes the visual image on the retina during head movement by counter-rotating the eyes in the head. The VOR is under adaptive control and has been extensively used to investigate sensorimotor transformations and motor learning. The cerebellar flocculus (FL) is intimately involved in VOR motor control and learning, because its sole output, the Purkinje cell firing pattern, modulates during visual-vestibular interaction paradigms that induce motor learning, and flocculectomy impairs the ability to modify the VOR. However, the role of the FL in VOR motor learning and the possibility of another neuronal site responsible for learning have been controversial. Currently, we performed single unit recordings of FL Purkinje cells during adaptation of the vertical (V) VOR in squirrel monkeys and used a system identification approach to localize the adaptable neuronal sites responsible for VVOR motor learning. We demonstrated that there are multiple adaptive sites: one upstream or in the FL, another in the non-FL pathway; change in the former site is in the wrong direction to cause the observed VVOR gain change, whereas that in the latter site is in the correct direction. The possibility that FL Purkinje cells convey an error signal to their target neurons that can be used to aid learning is discussed. PMID- 12582077 TI - The nonvisual complex spike signal in the flocculus responds to challenges to the vestibulo-ocular reflex gain. PMID- 12582078 TI - Role of caudal fastigial neurons during head-free gaze shifts in the monkey. PMID- 12582079 TI - Behavioral analysis of gain control for smooth pursuit eye movements. PMID- 12582080 TI - Coherence of subthreshold activity in coupled inferior olivary neurons. PMID- 12582081 TI - Comparing continuous and discrete movements with fMRI. PMID- 12582083 TI - What, indeed, do climbing fibers signal? Some cerebellar microcomplexes appear to monitor the activity in spinal nociceptive withdrawal reflex modules. PMID- 12582082 TI - Is the cerebellum important for podokinetic adaptation? PMID- 12582084 TI - Cell morphology and local circuitry of the central lobes of the mormyrid cerebellum. PMID- 12582085 TI - Evidence of a cerebrocerebellar network: an fMRI study of functional integration during visuospatiomotor sequence learning. PMID- 12582103 TI - Essential medicines twenty-five years on: closing the access gap. PMID- 12582105 TI - Transferring policies for treating sexually transmitted infections: what's wrong with global guidelines? AB - The paper uses a case study of the development of syndromic management for treating sexually transmitted infections (STIs) and subsequent policies recommending worldwide use of syndromic management guidelines. These treatment policies emerged in the late 1970s from researchers and public health physicians working in sub-Saharan Africa where they had to treat large numbers of STIs in difficult circumstances. Syndromic management was initially developed in specific local epidemiological and resource situations. By the late 1980s, the World Health Organization had adopted syndromic management as policy, and began to promote it globally in the form of algorithms and training guidelines. Dissemination was assisted by the context of the rapid spread of HIV/AIDS and the apparent effectiveness of syndromic management for treating STIs and slowing the transmission of HIV/AIDS. In the mid 1990s, international donors interested in HIV control and women's reproductive health took it up, and encouraged national programmes to adopt the new guidelines. Implementation, however, was a great deal more complex than anticipated, and was exacerbated by differences between three rather separate policy networks involved in the dissemination and execution of the global guidelines. The analysis focuses on two parts of the process of policy transfer: the organic development of scientific and medical consensus around a new policy for the treatment of STIs; and the formulation and subsequent dissemination of international policy guidelines. Using a political science approach, we analyze the transition from clinical tools to global guidelines, and the associated debates that accompanied their use. Finally, we comment on the way current global guidelines need to be adapted, given the growth in knowledge. PMID- 12582104 TI - Cost and cost-effectiveness of HIV/AIDS prevention strategies in developing countries: is there an evidence base? AB - Many donors and countries are striving to respond to the HIV/AIDS epidemic by implementing prevention programmes. However, the resources available for providing these activities relative to needs are limited. Hence, decision-makers must choose among various types of interventions. Cost information, both measures of cost and cost-effectiveness, serves as a critical input into the processes of setting priorities and allocating resources efficiently. This paper reviews the cost and cost-effectiveness evidence base of HIV/AIDS prevention programmes in low- and middle-income countries (LMICs). None of the studies found have complete cost data for a full range of HIV/AIDS prevention programmes in any one country. However, the range of studies highlight the relative emphasis of different types of HIV/AIDS prevention strategies by region, reflecting the various modes of transmission and hence, to a certain extent, the stage of the epidemic. The costing methods applied and results obtained in this review give rise to questions of reliability, validity and transparency. First, not all of the studies report the methods used to calculate the costs, and/or do not provide all the necessary data inputs such that recalculation of the results is possible. Secondly, methods that are documented vary widely, rendering different studies, even within the same country and programme setting, largely incomparable. Finally, even with consistent and replicable measurement, the results as presented are generally not comparable because of the lack of a common outcome measure. Therefore, the extent to which the available cost and cost-effectiveness evidence base on HIV/AIDS prevention strategies can provide guidance to decision makers is limited, and there is an urgent need for the generation of this knowledge for planning and decision-making. PMID- 12582106 TI - The SAZA study: implementing health financing reform in South Africa and Zambia. AB - This paper explores the policy-making process in the 1990s in two countries, South Africa and Zambia, in relation to health care financing reforms. While much of the analysis of health reform programmes has looked at design issues, assuming that a technically sound design is the primary requirement of effective policy change, this paper explores the political and bureaucratic realities shaping the pattern of policy change and its impacts. Through a case study approach, it provides a picture of the policy environment and processes in the two countries, specifically considering the extent to which technical analysts and technical knowledge were able to shape policy change. The two countries' experiences indicate the strong influence of political factors and actors over which health care financing policies were implemented, and which not, as well as over the details of policy design. Moments of political transition in both countries provided political leaders, specifically Ministers of Health, with windows of opportunity in which to introduce new policies. However, these transitions, and the changes in administrative structures introduced with them, also created environments that constrained the processes of reform design and implementation and limited the equity and sustainability gains achieved by the policies. Technical analysts, working either inside or outside government, had varying and often limited influence. In part, this reflected the limits of their own capacity as well as weaknesses in the way they were used in policy development. In addition, the analysts were constrained by the fact that their preferred policies often received only weak political support. Focusing almost exclusively on designing policy reforms, these analysts gave little attention to generating adequate support for the policy options they proposed. Finally, the country experiences showed that front-line health workers, middle level managers and the public had important influences over policy implementation and its impacts. The limited attention given to communicating policy changes to, or consulting with, these actors only heightened the potential for reforms to result in unanticipated and unwanted impacts. The strength of the paper lies in its 'thick description' of the policy process in each country, an empirical case study approach to policy that is under-represented in the literature. While such an approach allows only a cautious drawing of general conclusions, it suggests a number of ways in which to strengthen the implementation of financing policies in each country. PMID- 12582107 TI - A tale of two visions: the changing fortunes of Social Health Insurance in South Africa. AB - Over the last two decades there has been growing interest in the potential of Social Health Insurance (SHI) as a health care financing mechanism in low- and middle-income countries. SHI schemes exist in many countries of Latin America and have also been introduced across Asia in recent years. However, few countries in Africa have implemented SHI. Despite the sustained interest in SHI, there are growing concerns about whether it will achieve equity and financial sustainability objectives. It is clear that the design of SHI is of critical importance in this regard. South Africa has debated whether or not to implement SHI for nearly a decade and a half, and the design of SHI proposals has changed dramatically over that period. This paper considers lessons that can be drawn from the South African experience of developing and adapting SHI design, in conjunction with recent evidence from other low- and middle-income countries. The paper illustrates how SHI design may be affected by the trade-offs that are made to accommodate key actors' views, an accommodation that may even undermine the achievement of key objectives. A critical design requirement to promote equity and sustainability is a common contribution and risk pool across the SHI and any existing private insurers. In addition, given the complexity of SHI reforms, and the fact that SHI is usually only one component of a broader package of health sector reforms, the appropriate sequencing of implementation of the SHI and associated, supporting organizational and financing reforms is essential. As SHI evolves within a country, it is important to benchmark the changing nature of its design against pre-determined objectives in order to protect the final integrity of the policy. PMID- 12582108 TI - Health sector reform in the Occupied Palestinian Territories (OPT): targeting the forest or the trees? AB - Since the signing of the Oslo Peace Accords and the establishment of the Palestinian Authority in 1994, reform activities have targeted various spheres, including the health sector. Several international aid and UN organizations have been involved, as well as local and international non-governmental organizations, with considerable financial and technical investments. Although important achievements have been made, it is not evident that the quality of care has improved or that the most pressing health needs have been addressed, even before the second Palestinian Uprising that began in September 2000. The crisis of the Israeli re-invasion of Palestinian-controlled towns and villages since April 2002 and the attendant collapse of state structures and services have raised the problems to critical levels. This paper attempts to analyze some of the obstacles that have faced reform efforts. In our assessment, those include: ongoing conflict, frail Palestinian quasi-state structures and institutions, multiple and at times inappropriate donor policies and practices in the health sector, and a policy vacuum characterized by the absence of internal Palestinian debate on the type and direction of reform the country needs to take. In the face of all these considerations, it is important that reform efforts be flexible and consider realistically the political and economic contexts of the health system, rather than focus on mere narrow technical, managerial and financial solutions imported from the outside. PMID- 12582109 TI - Restructuring a ministry of health - an issue of structure and process: a case study from Uganda. AB - This paper analyzes the recently undertaken restructuring of the Ministry of Health in Uganda, which was carried out because the previous structure did not comply well with the decentralization framework or policy. The Ministry was put under pressure by higher political levels, supported by the donor community, to restructure. The paper describes the principles that were guiding this operation, and assesses whether these principles relate to the final outcome and whether the outcome was actually as expected. This paper also analyzes the reasons for the achieved, or not achieved, results and suggests the way forward. The outcome of the restructuring procedure in terms of change in number of employees was small. The foundation, in terms of a certain structure, has been laid for the Ministry of Health to function in its new role as a coach more than a player, but the work to establish a functioning process is far from complete. The issue is to make actors at various levels relate to each other, and thereby establish appropriate processes within the existing structure that will help the Ministry develop into a dynamic body. The importance of finding forms for interaction, involving face to-face relationships between the Ministry and service delivery level, is stressed. PMID- 12582110 TI - Improving government health services through contract management: a case from Cambodia. AB - Most government health facilities in Cambodia perform poorly, due to lack of funds, inadequate management and inefficient use of resources, but mostly due to poor motivation of staff. This paper describes contracting as a possible tool for Ministries of Health to improve health service delivery more rapidly than the more traditional reform approaches. In Cambodia, the Ministry of Health started an experiment with contracting in eight districts, covering 1 million people. Health care management in five districts was sub-contracted to private sector operators, and their results were compared with three control districts. Both internal and external reviews showed that after 3 years of implementation, the utilization of health services in the contracted districts improved significantly, in comparison with the control districts. There was adequate competition in awarding the contracts. A Ministry of Health Project Co-ordinating Unit measured the performance of the contractors, and contributed pro-actively. There was no evidence of rent-seeking practices by either the contracting agency or the contractors. This paper describes in more detail the successes and failures in one of the contracted districts, where HealthNet International applied the contracting approach. Despite significantly increased official user fees, constituting 16% of recurrent costs, the utilization of services was equally increased. Patients thought the fees were reasonable because they were still lower than the fees demanded if government health workers charged informally. They also thought that the services were of better quality than in the unregulated private sector. Another important result was that combining strict monitoring with performance-based incentives demonstrates a decrease in total family health expenditure of some 40% from US dollars 18 to US dollars 11 per capita per year. Innovative and decisive management proved to be essential, which is more likely to be achieved by a contracted manager than by regular government managers with life-long employment. This paper discusses how the contractor addressed the deeply rooted problems of informal private activities of government health workers. The NGO district management experimented with two management systems: first by individual contracts with health workers, and secondly by sub-contracting directly with the health centre chiefs and hospital directors. A reason for concern is that poli-pharmacy and excessive use of injectables continued. Also, the participation of the central level of the Ministry of Health was positive in the contracting process, but the role and participation of the provincial level of the Ministry was more tentative. PMID- 12582111 TI - Payment system reform for health care providers in Korea. AB - Since its introduction in 1977, the national health insurance programme in Korea has paid health care providers on a fee-for-service basis. Regulated fee-for service payment has resulted in an increased volume and intensity of medical care. It has also distorted the input mix of treatment because physicians have substituted more profitable and uninsured (no coverage) medical services for those with lower margins, as is evidenced by the sharp increase in the caesarean delivery rate. This paper examines two recent supply-side reforms in Korea: Diagnosis Related Group (DRG) and Resource-based Relative Value (RBRV). Since 1997, through a pilot programme covering a selected group of diseases for voluntarily participating health care institutions, the DRG-based prospective payment system has proven to be effective in containing cost with little negative effect on quality. RBRV-based payment was implemented in 2001, but led to an almost uniform increase in fees for physician services without a mechanism to control the volume and expenditure. Challenges and future issues in the reform of the payment system in Korea include the expansion of benefit coverage, quality monitoring and improvement, strategic plans to overcome the strong opposition of providers and the introduction of global budgeting. PMID- 12582112 TI - Fighting arsenic at the grassroots: experience of BRAC's community awareness initiative in Bangladesh. AB - The study evaluates the arsenic mitigation project of BRAC in raising awareness of arsenic poisoning in rural communities in Bangladesh. Data came from selected project villages in south-western Bangladesh. Comparison villages were also selected from the same region. A total of 1240 randomly selected adults were interviewed in May 2000. Findings reveal that the mitigation project played a significant, positive role in raising awareness of the safe water options, signs of arsenicosis, mode of transmission and the type of treatment. Testing tube-well water for arsenic created curiosity, innovation and interest in the community, and the water treatment plant became a symbol of the arsenic campaign. The study concludes that the behavioural change aspects of the arsenic mitigation project have the potential to significantly improve the level of understanding about arsenic contamination in the traditional communities. PMID- 12582113 TI - Factors associated with oral contraceptive discontinuation in rural Bangladesh. AB - Oral contraceptives (OCs) account for half of all modern contraceptive methods used in Bangladesh, however, discontinuation remains fairly high in OC use. This paper identifies factors associated with discontinuation of OC use, where discontinuation refers to cessation of OC use in the 6 months prior to the survey. The data for this study were drawn from a survey on OC compliance in rural Bangladesh. A total of 1600 OC users, current or past, aged 15 to 49 years were interviewed; of these, 36% discontinued OC use. Of the women who discontinued, 47% reported the experience of side-effects as the main reason for OC discontinuation. Multivariate analysis identified lack of fieldworker's visit as the strongest predictor of OC discontinuation; women who were not visited by fieldworkers had a four-fold risk of discontinuing OC use. Discontinuation of OC use decreased with increased duration of use and number of living children. OC discontinuation was associated with side-effect experiences, lack of husband's support in OC use and failure to purchase OCs. Of great concern is that about 70% of the women who were at risk of unintended pregnancy were not using any method of contraception following OC discontinuation. Husband's education was positively associated with the substitution of OCs with another contraceptive method. Effective OC use should be advocated through adequate counselling about how to take it correctly, the possibility of side-effects and their proper management and, more importantly, the possible alternative contraceptive method should OCs prove unsatisfactory or unsuitable. Better provider-client interactions along with improved access to the newly established community clinics could be instrumental in the continued and effective use of OCs. PMID- 12582114 TI - Social network analysis for health and social interventions among Kenyan scavenging street children. AB - Street children are a high priority for health policy and service planning in Kenya. Poverty, wars, famine and disease have resulted in street children having a persistent presence in African cities and towns. The Maastricht Social Network Analysis (MSNA) was implemented as the core instrument in a battery to measure the health status of the street children. Owing to the absence of census data of street children in Kenya and the difficulty in tracking this mobile population, we implemented a mixed snowball and convenience sampling design to recruit research subjects. Three hundred street and orphanage children, and 100 primary school children as a control group, were included in the study. The MSNA provided a social diagnosis that complements the clinical diagnosis of the health status of the sample. Only one main methodological question is presented: is the MSNA applicable to describe the personal social networks of (1). children and of (2). people living in a Kenyan culture? Qualitative field observations, key informant interviews and focus groups inform the adaptation of the MSNA instrument, and improve its face validity. A case series analysis is presented. The main result is that the street children population consists of distinct subgroups defined by the UNICEF classification as 'on' and 'of' the street and by gender. Street children networks have some notable deficiencies depending on the subgroup. Constant across the groups is the deficiency of service providers in their networks. The conclusion is that the MSNA is a suitable instrument for obtaining a social diagnosis and gathering other useful information that helps in understanding the social and health backgrounds, status and daily experiences of Kenyan scavenging street children. Applying the MSNA protocol was successful in the diagnosis and interpretation of the findings. PMID- 12582115 TI - Ten best readings in ... essential medicines. PMID- 12582117 TI - Fatal attraction: nonself recognition and heterokaryon incompatibility in filamentous fungi. PMID- 12582118 TI - Cdc24, the GDP-GTP exchange factor for Cdc42, is required for invasive hyphal growth of Candida albicans. AB - Candida albicans, the most common human fungal pathogen, is particularly problematic for immunocompromised individuals. The reversible transition of this fungal pathogen to a filamentous form that invades host tissue is important for its virulence. Although different signaling pathways such as a mitogen-activated protein kinase and a protein kinase A cascade are critical for this morphological transition, the function of polarity establishment proteins in this process has not been determined. We examined the role of four different polarity establishment proteins in C. albicans invasive growth and virulence by using strains in which one copy of each gene was deleted and the other copy expressed behind the regulatable promoter MET3. Strikingly, mutants with ectopic expression of either the Rho G-protein Cdc42 or its exchange factor Cdc24 are unable to form invasive hyphal filaments and germ tubes in response to serum or elevated temperature and yet grow normally as a budding yeast. Furthermore, these mutants are avirulent in a mouse model for systemic infection. This function of the Cdc42 GTPase module is not simply a general feature of polarity establishment proteins. Mutants with ectopic expression of the SH3 domain containing protein Bem1 or the Ras-like G-protein Bud1 can grow in an invasive fashion and are virulent in mice, albeit with reduced efficiency. These results indicate that a specific regulation of Cdc24/Cdc42 activity is required for invasive hyphal growth and suggest that these proteins are required for pathogenicity of C. albicans. PMID- 12582119 TI - Yap1 accumulates in the nucleus in response to carbon stress in Saccharomyces cerevisiae. AB - Yap1 is a transcription factor of the AP-1 family that is required for the adaptive response to oxidative stress in Saccharomyces cerevisiae. We recovered Yap1 in a two-hybrid screen for proteins that interact with the Sip2 subunit of the Snf1 protein kinase, which is required for the adaptation of cells to glucose limitation. Yap1 becomes enriched in the nucleus when cells are subjected to oxidative stress. We show that the localization of Yap1 is similarly sensitive to carbon stress. When glucose-grown cells were shifted to medium containing glycerol or no added carbon source, green fluorescent protein (GFP)-Yap1 accumulated in the nucleus. After adaptation to growth in glycerol, GFP-Yap1 was again primarily cytoplasmic. Nuclear accumulation was independent of respiration and of the Snf1, PKA, TOR, and Yak1 pathways, and the mechanism is distinct from that involved in the response to hydrogen peroxide. Addition of glutathione to the medium inhibited nuclear accumulation of GFP-Yap1 in response to carbon stress but did not affect the relocalization of Gal83 or Mig1. Other stresses such as increased temperature, acidic pH, and ionic stress did not cause nuclear enrichment of GFP-Yap1. These findings suggest a role for Yap1 in the response to carbon stress. PMID- 12582120 TI - Ssk1p response regulator binding surface on histidine-containing phosphotransfer protein Ypd1p. AB - Ypd1p, a histidine-containing phosphotransfer protein, plays an important role in a branched His-Asp phosphorelay signal transduction pathway that regulates cellular responses to hyperosmotic stress in Saccharomyces cerevisiae. Ypd1p is required for phosphoryl group transfer from the membrane-bound Sln1p sensor histidine kinase to two downstream response regulator proteins, Ssk1p and Skn7p. To investigate the molecular basis for interaction of Ypd1p with these response regulator domains, we used an approach that coupled alanine-scanning mutagenesis of surface-exposed residues in Ypd1p with a yeast two-hybrid interaction screen. Mutated residues that adversely affected the interaction of Ypd1p with the C terminal response regulator domain of Ssk1p were identified and found to cluster on or near the alphaA helix in Ypd1p. Our results, supported by analysis of a modeled complex, identify a binding site on Ypd1p for response regulators that is composed of a cluster of conserved hydrophobic residues surrounded by less conserved polar residues. We propose that molecular interactions involving Ypd1p are mediated primarily through hydrophobic contacts, whereas binding specificity and strength of interaction may be influenced by select polar side chain interactions. PMID- 12582121 TI - Expressed sequence tag analysis of the human pathogen Paracoccidioides brasiliensis yeast phase: identification of putative homologues of Candida albicans virulence and pathogenicity genes. AB - Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of the prevalent systemic mycosis in Latin America, paracoccidioidomycosis. We present here a survey of expressed genes in the yeast pathogenic phase of P. brasiliensis. We obtained 13,490 expressed sequence tags from both 5' and 3' ends. Clustering analysis yielded the partial sequences of 4,692 expressed genes that were functionally classified by similarity to known genes. We have identified several Candida albicans virulence and pathogenicity homologues in P. brasiliensis. Furthermore, we have analyzed the expression of some of these genes during the dimorphic yeast-mycelium-yeast transition by real-time quantitative reverse transcription-PCR. Clustering analysis of the mycelium-yeast transition revealed three groups: (i) RBT, hydrophobin, and isocitrate lyase; (ii) malate dehydrogenase, contigs Pb1067 and Pb1145, GPI, and alternative oxidase; and (iii) ubiquitin, delta-9-desaturase, HSP70, HSP82, and HSP104. The first two groups displayed high mRNA expression in the mycelial phase, whereas the third group showed higher mRNA expression in the yeast phase. Our results suggest the possible conservation of pathogenicity and virulence mechanisms among fungi, expand considerably gene identification in P. brasiliensis, and provide a broader basis for further progress in understanding its biological peculiarities. PMID- 12582122 TI - Cell biology of mating in Candida albicans. AB - It was recently demonstrated that strains homozygous for either of the mating type-like loci MTLa and MTLalpha of Candida albicans undergo white-opaque switching and that expression of the opaque-phase phenotype greatly enhances mating between strains. Exploiting the latter property to obtain high-frequency mating, we have characterized the cell biology of the mating process of C. albicans. Employing continuous videomicroscopy, computer-assisted three dimensional reconstruction of living cells, and fluorescence microscopy, we have monitored the mating-associated processes of conjugation, tube formation, fusion, budding, septum formation, and daughter cell development and the spatial and temporal dynamics of nuclear migration and division. From these observations, a model for the stages in C. albicans mating is formulated. The stages include shmooing, chemotropism of conjugation tubes, fusion of tubes and nuclear association, vacuole expansion and nuclear separation in the conjugation bridge, asynchronous nuclear division in the zygote, bud growth, nuclear migration into the daughter cell, septation, and daughter cell budding. Since there was no cytological indication of karyogamy, genetic experiments were performed to assess marker segregation. Recombination was not observed, suggesting that mating takes place in the absence of karyogamy between naturally occurring, homozygous a and alpha strains. This study provides the first description of the cell biology of the mating process of C. albicans. PMID- 12582123 TI - Constitutively active protein kinase A disrupts motility and chemotaxis in Dictyostelium discoideum. AB - The deletion of the gene for the regulatory subunit of protein kinase A (PKA) results in constitutively active PKA in the pkaR mutant. To investigate the role of PKA in the basic motile behavior and chemotaxis of Dictyostelium discoideum, pkaR mutant cells were subjected to computer-assisted two- and three-dimensional motion analysis. pkaR mutant cells crawled at only half the speed of wild-type cells in buffer, chemotaxed in spatial gradients of cyclic AMP (cAMP) but with reduced efficiency, were incapable of suppressing lateral pseudopods in the front of temporal waves of cAMP, a requirement for natural chemotaxis, did not exhibit the normal velocity surge in response to the front of a wave, and were incapable of chemotaxing toward an aggregation center in natural waves generated by wild type cells that made up the majority of cells in mixed cultures. Many of the behavioral defects appeared to be the result of the constitutively ovoid shape of the pkaR mutant cells, which forced the dominant pseudopod off the substratum and to the top of the cell body. The behavioral abnormalities that pkaR mutant cells shared with regA mutant cells are discussed by considering the pathway ERK2 perpendicular RegA perpendicular [cAMP] --> PKA, which emanates from the front of a wave. The results demonstrate that cells must suppress PKA activity in order to elongate along a substratum, suppress lateral-pseudopod formation, and crawl and chemotax efficiently. The results also implicate PKA activation in dismantling cell polarity at the peak and in the back of a natural cAMP wave. PMID- 12582124 TI - Glycosylphosphatidylinositol-dependent protein trafficking in bloodstream stage Trypanosoma brucei. AB - We have previously demonstrated that glycosylphosphatidylinositol (GPI) anchors strongly influence protein trafficking in the procyclic insect stage of Trypanosoma brucei (M. A. McDowell, D. A. Ransom, and J. D. Bangs, Biochem. J. 335:681-689, 1998), where GPI-minus variant surface glycoprotein (VSG) reporters have greatly reduced rates of endoplasmic reticulum (ER) exit but are ultimately secreted. We now demonstrate that GPI-dependent trafficking also occurs in pathogenic bloodstream trypanosomes. However, unlike in procyclic trypanosomes, truncated VSGs lacking C-terminal GPI-addition signals are not secreted but are mistargeted to the lysosome and degraded. Failure to export these reporters is not due to a deficiency in secretion of these cells since the N-terminal ATPase domain of the endogenous ER protein BiP is efficiently secreted from transgenic cell lines. Velocity sedimentation experiments indicate that GPI-minus VSG dimerizes similarly to wild-type VSG, suggesting that degradation is not due to ER quality control mechanisms. However, GPI-minus VSGs are fully protected from degradation by the cysteine protease inhibitor FMK024, a potent inhibitor of the major lysosomal protease trypanopain. Immunofluorescence of cells incubated with FMK024 demonstrates that GPI-minus VSG colocalizes with p67, a lysosomal marker. These data suggest that in the absence of a GPI anchor, VSG is mistargeted to the lysosome and subsequently degraded. Our findings indicate that GPI-dependent transport is a general feature of secretory trafficking in both stages of the life cycle. A working model is proposed in which GPI valence regulates progression in the secretory pathway of bloodstream stage trypanosomes. PMID- 12582125 TI - Targeting the variable surface of African trypanosomes with variant surface glycoprotein-specific, serum-stable RNA aptamers. AB - African trypanosomes cause sleeping sickness in humans and Nagana in cattle. The parasites multiply in the blood and escape the immune response of the infected host by antigenic variation. Antigenic variation is characterized by a periodic change of the parasite protein surface, which consists of a variant glycoprotein known as variant surface glycoprotein (VSG). Using a SELEX (systematic evolution of ligands by exponential enrichment) approach, we report the selection of small, serum-stable RNAs, so-called aptamers, that bind to VSGs with subnanomolar affinity. The RNAs are able to recognize different VSG variants and bind to the surface of live trypanosomes. Aptamers tethered to an antigenic side group are capable of directing antibodies to the surface of the parasite in vitro. In this manner, the RNAs might provide a new strategy for a therapeutic intervention to fight sleeping sickness. PMID- 12582126 TI - Selection on the genes of Euplotes crassus Tec1 and Tec2 transposons: evolutionary appearance of a programmed frameshift in a Tec2 gene encoding a tyrosine family site-specific recombinase. AB - The Tec1 and Tec2 transposons of the ciliate Euplotes crassus carry a gene for a tyrosine-type site-specific recombinase. The expression of the Tec2 gene apparently uses a programmed +1 frameshift. To test this hypothesis, we first examined whether this gene has evolved under purifying selection in Tec1 and Tec2. Each element carries three genes, and each has evolved under purifying selection for the function of its encoded protein, as evidenced by a dearth of nonsynonymous changes. This distortion of divergence is apparent in codons both 5' and 3' of the frameshift site. Thus, Tec2 transposons have diverged from each other while using a programmed +1 frameshift to produce recombinase, the function of which is under purifying selection. What might this function be? Tyrosine-type site-specific recombinases are extremely rare in eukaryotes, and Tec elements are the first known eukaryotic type II transposons to encode a site-specific recombinase. Tec elements also encode a widespread transposase. The Tec recombinase might function in transposition, resolve products of transposition (bacterial replicative transposons use recombinase or resolvase to separate joined replicons), or provide a function that benefits the ciliate host. Transposons in ciliated protozoa are removed from the macronucleus, and it has been proposed that the transposons provide this "excisase" activity. PMID- 12582127 TI - Tec3, a new developmentally eliminated DNA element in Euplotes crassus. AB - More than 100,000 interstitial segments of DNA (internal eliminated sequences [IESs]) are excised from the genome during the formation of a new macronucleus in Euplotes crassus. IESs include unique sequence DNA as well as two related families of transposable elements, Tec1 and Tec2. Here we describe a new class of E. crassus transposons, Tec3, which is present in 20 to 30 copies in the micronuclear genome. Tec3 elements have long inverted terminal repeats and contain a degenerate open reading frame encoding a tyrosine-type recombinase. One characterized copy of Tec3 (Tec3-1) is 4.48 kbp long, has 1.23-kbp inverted terminal repeats, and resides within the micronuclear copy of the ribosomal protein L29 gene (RPL29). The 23 bp at the extreme ends of this element are very similar to those in other E. crassus IESs and, like these other IESs, Tec3-1 is excised during the polytene chromosome stage of macronuclear development to generate a free circular form with an unusual junction structure. In contrast, a second cloned element, Tec3-2, is quite similar to Tec3-1 but lacks the terminal 258 bp of the inverted repeats, so that its ends do not resemble the other E. crassus IES termini. The Tec3-2 element appears to reside in a large segment of the micronuclear genome that is subject to developmental elimination. Models for the origins of these two types of Tec3 elements are presented, along with a discussion of how some members of this new transposon family may have come to be excised by the same machinery that removes other E. crassus IESs. PMID- 12582128 TI - Origin usage during euplotes ribosomal DNA amplification. AB - The macronuclear genome of the ciliate Euplotes is comprised of millions of small linear DNA molecules that have telomeres on each end. These molecules are generated during the sexual stage of the life cycle, when the new macronucleus is formed by a series of DNA processing events and multiple rounds of DNA amplification. We have used two-dimensional gels to compare the location of the replication origins used during vegetative growth and the two periods during macronuclear development when DNA amplification takes place. When we examined the pattern of ribosomal DNA (rDNA) replication intermediates, we observed almost identical Y arcs regardless of when in the Euplotes life cycle the DNA was isolated. No bubble or bubble-to-Y arcs could be detected. This indicates that replication of the macronuclear rDNA initiates at or near the telomere even when these molecules are being differentially amplified. Since replication rarely initiated from both ends of the rDNA, we examined the direction of replication fork movement to determine which end of the rDNA served as the origin. Fork movement gels indicated that replication initiated at the 5' end. As transcription also starts near the telomere at the 5' end, our findings suggest that the telomere and the promoter region cooperate to recruit Euplotes replication initiation complexes. PMID- 12582129 TI - Overproduction of polypeptides corresponding to the amino terminus of the F-box proteins Cdc4p and Met30p inhibits ubiquitin ligase activities of their SCF complexes. AB - Ubiquitin ligases direct the transfer of ubiquitin onto substrate proteins and thus target the substrate for proteasome-dependent degradation. SCF complexes are a family of ubiquitin ligases composed of a common core of components and a variable component called an F-box protein that defines substrate specificity. Distinct SCF complexes, defined by a particular F-box protein, target different substrate proteins for degradation. Although a few have been identified to be involved in important biological pathways, such as the cell division cycle and coordinating cellular responses to changes in environmental conditions, the role of the overwhelming majority of F-box proteins is not clear. Creating inhibitors that will block the in vivo activities of specific SCF ubiquitin ligases may provide identification of substrates of these uncharacterized F-box proteins. Using Saccharomyces cerevisiae as a model system, we demonstrate that overproduction of polypeptides corresponding to the amino terminus of the F-box proteins Cdc4p and Met30p results in specific inhibition of their SCF complexes. Analyses of mutant amino-terminal alleles demonstrate that the interaction of these polypeptides with their full-length counterparts is an important step in the inhibitory process. These results suggest a common means to inhibit specific SCF complexes in vivo. PMID- 12582130 TI - mRNAs encoding telomerase components and regulators are controlled by UPF genes in Saccharomyces cerevisiae. AB - Telomeres, the chromosome ends, are maintained by a balance of activities that erode and replace the terminal DNA sequences. Furthermore, telomere-proximal genes are often silenced in an epigenetic manner. In Saccharomyces cerevisiae, average telomere length and telomeric silencing are reduced by loss of function of UPF genes required in the nonsense-mediated mRNA decay (NMD) pathway. Because NMD controls the mRNA levels of several hundred wild-type genes, we tested the hypothesis that NMD affects the expression of genes important for telomere functions. In upf mutants, high-density oligonucleotide microarrays and Northern blots revealed that the levels of mRNAs were increased for genes encoding the telomerase catalytic subunit (Est2p), in vivo regulators of telomerase (Est1p, Est3p, Stn1p, and Ten1p), and proteins that affect telomeric chromatin structure (Sas2p and Orc5p). We investigated whether overexpressing these genes could mimic the telomere length and telomeric silencing phenotypes seen previously in upf mutant strains. Increased dosage of STN1, especially in combination with increased dosage of TEN1, resulted in reduced telomere length that was indistinguishable from that in upf mutants. Increased levels of STN1 together with EST2 resulted in reduced telomeric silencing like that of upf mutants. The half-life of STN1 mRNA was not altered in upf mutant strains, suggesting that an NMD-controlled transcription factor regulates the levels of STN1 mRNA. Together, these results suggest that NMD maintains the balance of gene products that control telomere length and telomeric silencing primarily by maintaining appropriate levels of STN1, TEN1, and EST2 mRNA. PMID- 12582131 TI - Glucose regulation of Saccharomyces cerevisiae cell cycle genes. AB - Nutrient-limited Saccharomyces cerevisiae cells rapidly resume proliferative growth when transferred into glucose medium. This is preceded by a rapid increase in CLN3, BCK2, and CDC28 mRNAs encoding cell cycle regulatory proteins that promote progress through Start. We have tested the ability of mutations in known glucose signaling pathways to block glucose induction of CLN3, BCK2, and CDC28. We find that loss of the Snf3 and Rgt2 glucose sensors does not block glucose induction, nor does deletion of HXK2, encoding the hexokinase isoenzyme involved in glucose repression signaling. Rapamycin blockade of the Tor nutrient sensing pathway does not block the glucose response. Addition of 2-deoxy glucose to the medium will not substitute for glucose. These results indicate that glucose metabolism generates the signal required for induction of CLN3, BCK2, and CDC28. In support of this conclusion, we find that addition of iodoacetate, an inhibitor of the glyceraldehyde-3-phosphate dehydrogenase step in yeast glycolysis, strongly downregulates the levels CLN3, BCK2, and CDC28 mRNAs. Furthermore, mutations in PFK1 and PFK2, which encode phosphofructokinase isoforms, inhibit glucose induction of CLN3, BCK2, and CDC28. These results indicate a link between the rate of glycolysis and the expression of genes that are critical for passage through G(1). PMID- 12582132 TI - Transcriptional regulation of xyn2 in Hypocrea jecorina. AB - The xylanase system of the filamentous fungus Hypocrea jecorina (Trichoderma reesei) consists of two specific xylanases, Xyn1 and Xyn2, which are simultaneously expressed during growth on xylan but respond differentially to low molecular-weight inducers. Using in vivo footprinting analysis of xylan-induced and noninduced mycelia, we detected two adjacent nucleotide sequences (5'-AGAA-3' on the noncoding strand and 5'-GGGTAAATTGG-3', referred to as the xylanase activating element [XAE], on the coding strand, respectively) to bind proteins. Among these, binding to the AGAA-box is only observed under noninduced conditions, whereas binding to XAE is constitutive. Electrophoretic mobility shift assay with heterologously expressed components of the H. jecorina Hap2/3/5 protein complex and the cellulase regulator Ace2 suggests that these two transactivators form the protein complex binding to XAE. H. jecorina transformants, containing correspondingly mutated versions of the xyn2 promoter fused to the Aspergillus niger goxA gene as a reporter, revealed that the elimination of protein binding to the AGAA-box resulted in a threefold increase in both basal and induced transcription, whereas elimination of Ace2 binding to its target in XAE completely eliminated transcription under both conditions. Destruction of the CCAAT-box by insertion of a point mutation prevents binding of the Hap2/3/5 complex in vitro and results in a slight increase in both basal and induced transcription. These data support a model of xyn2 regulation based on the interplay of Hap2/3/5, Ace2 and the AGAA-box binding repressor. PMID- 12582133 TI - Bgs3p, a putative 1,3-beta-glucan synthase subunit, is required for cell wall assembly in Schizosaccharomyces pombe. AB - beta-Glucans are the main components of the fungal cell wall. Fission yeast possesses a family of beta-glucan synthase-related genes. We describe here the cloning and characterization of bgs3(+), a new member of this family. bgs3(+) was cloned as a suppressor of a mutant hypersensitive to Echinocandin and Calcofluor White, drugs that interfere with cell wall biosynthesis. Disruption of the gene is lethal, and a decrease in Bgs3p levels leads to rounded cells with thicker walls, slightly reduces the amount of the beta-glucan, and raises the amount of alpha-glucan polymer. These cells finally died. bgs3(+) is expressed in vegetative cells grown in different conditions and during mating and germination and is not enhanced by stress situations. Consistent with the observed expression pattern, Bgs3-green fluorescence protein (GFP-Bgs3p) was found at the growing tips during interphase and at the septum prior to cytokinesis, always localized to growth areas. We also found GFP-Bgs3p in mating projections, during the early stages of zygote formation, and at the growing pole during ascospore germination. We conclude that Bgs3p localization is restricted to growth areas and that Bgs3p is a glucan synthase homologue required for cell wall biosynthesis and cell elongation in the fission yeast life cycle. PMID- 12582134 TI - Metabolic-flux profiling of the yeasts Saccharomyces cerevisiae and Pichia stipitis. AB - The so far largely uncharacterized central carbon metabolism of the yeast Pichia stipitis was explored in batch and glucose-limited chemostat cultures using metabolic-flux ratio analysis by nuclear magnetic resonance. The concomitantly characterized network of active metabolic pathways was compared to those identified in Saccharomyces cerevisiae, which led to the following conclusions. (i) There is a remarkably low use of the non-oxidative pentose phosphate (PP) pathway for glucose catabolism in S. cerevisiae when compared to P. stipitis batch cultures. (ii) Metabolism of P. stipitis batch cultures is fully respirative, which contrasts with the predominantly respiro-fermentative metabolic state of S. cerevisiae. (iii) Glucose catabolism in chemostat cultures of both yeasts is primarily oxidative. (iv) In both yeasts there is significant in vivo malic enzyme activity during growth on glucose. (v) The amino acid biosynthesis pathways are identical in both yeasts. The present investigation thus demonstrates the power of metabolic-flux ratio analysis for comparative profiling of central carbon metabolism in lower eukaryotes. Although not used for glucose catabolism in batch culture, we demonstrate that the PP pathway in S. cerevisiae has a generally high catabolic capacity by overexpressing the Escherichia coli transhydrogenase UdhA in phosphoglucose isomerase-deficient S. cerevisiae. PMID- 12582135 TI - TOK homologue in Neurospora crassa: first cloning and functional characterization of an ion channel in a filamentous fungus. AB - In contrast to animal and plant cells, very little is known of ion channel function in fungal physiology. The life cycle of most fungi depends on the "filamentous" polarized growth of hyphal cells; however, no ion channels have been cloned from filamentous fungi and comparatively few preliminary recordings of ion channel activity have been made. In an attempt to gain an insight into the role of ion channels in fungal hyphal physiology, a homolog of the yeast K(+) channel (ScTOK1) was cloned from the filamentous fungus, Neurospora crassa. The patch clamp technique was used to investigate the biophysical properties of the N. crassa K(+) channel (NcTOKA) after heterologous expression of NcTOKA in yeast. NcTOKA mediated mainly time-dependent outward whole-cell currents, and the reversal potential of these currents indicated that it conducted K(+) efflux. NcTOKA channel gating was sensitive to extracellular K(+) such that channel activation was dependent on the reversal potential for K(+). However, expression of NcTOKA was able to overcome the K(+) auxotrophy of a yeast mutant missing the K(+) uptake transporters TRK1 and TRK2, suggesting that NcTOKA also mediated K(+) influx. Consistent with this, close inspection of NcTOKA-mediated currents revealed small inward K(+) currents at potentials negative of E(K). NcTOKA single channel activity was characterized by rapid flickering between the open and closed states with a unitary conductance of 16 pS. NcTOKA was effectively blocked by extracellular Ca(2+), verapamil, quinine, and TEA(+) but was insensitive to Cs(+), 4-aminopyridine, and glibenclamide. The physiological significance of NcTOKA is discussed in the context of its biophysical properties. PMID- 12582136 TI - MsGC-beta3 forms active homodimers and inactive heterodimers with NO-sensitive soluble guanylyl cyclase subunits. AB - Soluble guanylyl cyclases are typically obligate heterodimers, composed of a single alpha and a single beta subunit. MsGC-beta3, identified in the tobacco hornworm Manduca sexta, was the first example of a soluble guanylyl cyclase that exhibited enzyme activity without the need for coexpression with additional subunits. Subsequent studies have revealed that the mammalian beta2 subunit also shares this property. Using a combination of gel filtration chromatography, coprecipitation and site-directed mutagenesis we show that, as predicted, MsGC beta3 forms active homodimers. We also demonstrate that MsGC-beta3 is capable of forming heterodimers with the nitric oxide (NO)-sensitive guanylyl cyclase subunits MsGC-alpha1 and MsGC-beta1. These heterodimers, however, show no enzyme activity and, like mammalian beta2 subunits, act in a dominant negative manner when combined with the NO-sensitive subunits to disrupt their activation by NO. In addition, we show that the unique C-terminal domain of MsGC-beta3 is not necessary for enzyme activity and might act as an auto-inhibitory domain. PMID- 12582137 TI - K(+) transport in Malpighian tubules of Tenebrio molitor L: a study of electrochemical gradients and basal K(+) uptake mechanisms. AB - Malpighian tubules of the mealworm Tenebrio molitor were isolated for intracellular measurement of basolateral (V(bl)) and, indirectly, apical (V(ap)) membrane potentials. In control Ringer (50 mmol l(-1) K(+), 140 mmol l(-1) Na(+)), V(bl) was 24 mV, cell negative, and V(ap) was 48 mV, cell negative with reference to the lumen. Ion substitution experiments involving K(+) and Na(+) indicated that both V(bl) and V(ap) were sensitive to the bathing K(+) concentration, with the change in V(ap) being 60-77% that of V(bl). A 10-fold drop in bath [K(+)] irreversibly decreased fluid secretion rates from 6.38+/-0.95 nl x min(-1) (mean +/- S.E.M.) to 1.48+/-0.52 nl x min(-1) (N=8). In the presence of 6 mmol l(-1) Ba(2+), a blocker of basal K(+) channels, fluid secretion rates reversibly decreased and the hyperpolarization of both V(bl) and V(ap) seen in 50 mmol l(-1) and 140 mmol l(-1) K(+) indicated a favourable electrochemical gradient for basal K(+) entry. In 5 mmol l(-1) K(+), Ba(2+) induced two different responses: V(bl) either hyperpolarized by approximately 10 mV or depolarised by approximately 14 mV, according to the electrochemical gradient for K(+), which was either inward or outward in low bath [K(+)]. Rubidium, a 'permeant' potassium substitute, caused a hyperpolarization of V(bl), indicating the specificity of K(+) channels found in Tenebrio tubule cells. Other possible K(+) uptake mechanisms located in the basolateral membrane were investigated. Blocking of the putative electroneutral Na(+)/K(+)/2Cl(-) cotransporter by 10 micromol l(-1) bumetanide reversibly decreased fluid secretion rates, with no detectable change in membrane potentials. Ouabain (1 mmol l(-1)), an Na(+)/K(+)-ATPase inhibitor, irreversibly decreased fluid secretion rates but had no effect on electrical potential differences either in the absence or presence of Ba(2+). The results implicate K(+) channels, the Na(+)/K(+)/2Cl(-) contransporter and the Na(+)/K(+) ATPase in basal K(+) and fluid transport of Tenebrio tubule cells. PMID- 12582138 TI - K(+) transport in Malpighian tubules of Tenebrio molitor L: is a K(ATP) channel involved? AB - The presence of ATP-regulated K(+) (K(ATP)) channels in Tenebrio molitor Malpighian tubules was investigated by examining the effect of glibenclamide on both fluid secretion and basolateral membrane potentials (V(bl)). Glibenclamide, a K(ATP) channel blocker, slowed fluid secretion of Tenebrio tubules. In low bath K(+) concentration (5 mmol l(-1)), glibenclamide either hyperpolarized or depolarized V(bl), resembling the effect seen with Ba(2+). Subsequent addition of 6 mmol l(-1) Ba(2+) caused a further hyper- or depolarization of V(bl). In control Ringer (50 mmol l(-1) KCl, 90 mmol l(-1) NaCl), glibenclamide had no visible effect on V(bl). The effect of ouabain was investigated in low bath [K(+)] in the presence of Ba(2+). V(bl) responded by a small but significant hyperpolarization from -51+/-4 mV to -56+/-4 mV (n=16, P<0.001) in response to 1 mmol l(-1) ouabain. Repeating the experiments in the presence of both glibenclamide and Ba(2+) resulted in a depolarization of V(bl) when ouabain was added. In low bath [K(+)] (high Na(+)), the Na(+)/K(+)-ATPase is expected to function at a high rate. In the presence of Ba(2+), replacing Na(+) by K(+) rapidly depolarized V(bl), but this was followed by a repolarization. Repeating the experiments in the presence of glibenclamide markedly reduced the depolarizing effect and abolished the repolarization, with a gradual decrease in the sensitivity of V(bl) to the surrounding [K(+)]. These results suggest the presence of K(ATP) channels in the basolateral membrane. Glibenclamide had no visible effect on V(bl) in high K(+) or in the absence of Ba(2+), indicating that other highly conductive K(+) channels may mask the effect on K(ATP) channels. This is the first demonstration of the presence of K(ATP) channels in an insect epithelium. PMID- 12582139 TI - Respiratory patterns and oxygen consumption in singing zebra finches. AB - Song production in birds is driven by temporally complex respiratory patterns. In zebra finches (Taeniopygia guttata), song consists of repetitions of a stereotyped sequence of distinct syllables (motif). Syllables correspond to distinct expiratory pulses, which alternate with short deep inspirations. We investigated the effect of the song motor pattern on respiration using a newly developed mask system to measure oxygen consumption while simultaneously monitoring subsyringeal air sac pressure. The metabolic cost of song is closely related to song duration (mean=85.7 microl O(2) g(-1) x min(-1) over pre-song levels) and confirms earlier estimates for this species. When motif duration is controlled for, there is only small interindividual variability in oxygen consumption per motif. The oxygen measurements were also used to evaluate various methods of estimating and reporting the metabolic cost. Up to 20s before song, respiratory activity and oxygen consumption increased. Shortly before and during the introductory notes of the song bout, respiration and oxygen consumption decreased markedly. In some individuals, significant hyperventilation occurred during song, causing almost complete apnea after the song. In three different birds, we measured tracheal airflow and air sac pressure during song. Birds with higher airflow during song relative to flow during quiet respiration had a more pronounced decrease in respiratory activity after the song bout. These results suggest that gas exchange continues in the lung during the song bout and that long expiratory pressure pulses of the song motif can lead to hyperventilation but that no oxygen debt resulted from song. This research allows a first assessment of respiratory constraints that may influence the evolution of song complexity. PMID- 12582140 TI - Putative involvement of crustacean hyperglycemic hormone isoforms in the neuroendocrine mediation of osmoregulation in the crayfish Astacus leptodactylus. AB - This study investigates the involvement of eyestalk neuroendocrine factors on osmoregulation in the crayfish Astacus leptodactylus maintained in freshwater. Eyestalk removal was followed by a significant decrease in hemolymph osmolality and Na(+) concentration and by a 50% increase in mass after one molting cycle. Several neurohormones have been isolated from the sinus gland through high performance liquid chromatography (HPLC), and different crustacean hyperglycemic hormone (CHH)-related peptides, including stereoisomers (L-CHH and D-Phe(3) CHH), have been identified by direct enzyme-linked immunosorbent assay (ELISA). A glucose quantification bioassay demonstrated a strong hyperglycemic activity following injection of the immunoreactive chromatographic fractions and showed that the D-Phe(3) CHH was the most efficient. Destalked crayfish were then injected with purified CHH HPLC fractions. The D-Phe(3) CHH fraction significantly increased the hemolymph osmolality and Na(+) content 24 h after injection. Two other CHH-related peptides caused a smaller increase in Na(+) concentration. No significant variation was observed in hemolymph Cl(-) concentration following injection of any of the CHH isoforms. These results constitute the first observation of the effects of a CHH isoform, specifically the D-Phe(3) CHH, on osmoregulatory parameters in a freshwater crustacean. The effects of eyestalk ablation and CHH injection on osmoregulation and the identification of different CHH-related peptides and isoforms in crustaceans are discussed. PMID- 12582141 TI - Electroreception in G carapo: detection of changes in waveform of the electrosensory signals. AB - Electric fish evaluate the near environment by detecting changes in their self generated electric organ discharge. To investigate impedance modulation of the self-generated electric field, this field was measured at the electrosensory fovea of Gymnotus carapo in the presence and absence of objects. Changes in local fields provoked by resistive objects were predicted by the change in total energy. Objects with capacitive impedance generated large variations in the relative importance of the different waveform components of the electric organ discharge. We tested the hypothesis that fish discriminate changes in waveform as well as increases in total energy using the novelty response, which is a behavioural response consisting of a transient acceleration of EOD frequency that can follow a change in object impedance. For resistive loads, the amplitude of novelty responses was well predicted by the increase in total energy. For complex loads, the amplitude of novelty responses was correlated not only with increases in total energy but also with waveform changes, consisting of reductions in the early slow negative wave and increases in the late sharp negative wave. The total energy and waveform effects appeared to be additive. These results indicate that G. carapo discriminates complex impedance based on an evaluation of different waveform parameters. PMID- 12582142 TI - Probability and amplitude of novelty responses as a function of the change in contrast of the reafferent image in G carapo. AB - Pulse electric fish evaluate successive electrosensory images generated by self emitted electric discharges, creating a neural representation of the physical world. Intervals between discharges (system resolution) are controlled by a pacemaker nucleus under the influence of reafferent signals. Novel sensory stimuli cause transient accelerations of the pacemaker rate (novelty responses). This study describes quantitatively the effect of changes in contrast of reafferent electrosensory signals on the amplitude and probability of novelty responses. We found that: (i). alterations of a single image in an otherwise homogeneous series cause a novelty response; (ii). the amplitude of the elicited novelty response is a linear function of the logarithm of the change in image contrast; (iii). the parameters of this function, threshold and proportionality constant, allowed us to evaluate the transference function between change in stimulus amplitude and the amplitude of the novelty response; (iv). both parameters are independent of the baseline contrast; (v). the proportionality constant increases with the moving average of the contrast of hundreds of previous images. These findings suggest that the electrosensory system (i). calculates the difference between each reafferent electrosensory image and a neural representation of the past electrosensory input ('template'); (ii). creates the comparison template in which the relative contribution of every image decreases with the incorporation of successive images. We conclude that contrast discrimination in the electrosensory system of G. carapo obeys the general principle of appreciating any instantaneous input by the input's departure from a moving average of past images. PMID- 12582143 TI - Myogenic cell cycle duration in Harpagifer species with sub-Antarctic and Antarctic distributions: evidence for cold compensation. AB - In teleosts, the proliferation of myogenic progenitor cells is required for muscle growth and nuclear turnover. We measured the cell cycle and S-phase duration of myogenic cells in the fast myotomal muscle of two closely related Harpagifer species by cumulative S-phase labelling with 5-bromo-2'-deoxyuridine (BrdU). Harpagifer antarcticus is a stenothermal species from the Antarctic peninsula (experiencing temperatures of -2 degrees C to +1 degrees C) and Harpagifer bispinis is a eurythermal species from the Beagle Channel, Tierra del Fuego (living at +4 degrees C in winter and up to 11 degrees C in summer). Specific growth rates in the adult stages studied were not significantly different from zero. Myogenic progenitor cells were identified using an antibody against c-met. Seventy-five percent of the c-met(+ve) cells were in a proliferative state in both species. Cell cycle time was 150 h at 5 degrees C and 81.3 h at 10 degrees C in H. bispinis (Q(10)=3.4). Cell cycle duration was 35% shorter in H. antarcticus at 0 degrees C (111 h) than in H. bispinis at 5 degrees C. The predicted cell cycle time for H. bispinis at 0 degrees C (based on the Q(10) relationship) was 277 h, which was more than double that measured for the Antarctic species at this temperature. The results obtained are compatible with an evolutionary adjustment of cell cycle time for function at low temperature in the Antarctic species. PMID- 12582144 TI - Active NaCl absorption across posterior gills of hyperosmoregulating Chasmagnathus granulatus. AB - Split lamellae of posterior gills of Chasmagnathus granulatus adapted to 2.5 per thousand salinity were mounted in a modified Ussing chamber. With NaCl-saline on both sides of the preparation a transepithelial voltage (V(te)) of 4.1+/-0.5 mV (outside positive) was measured. After voltage-clamping, the negative short circuit current (I(sc)) amounted to -142+/-21 micro A cm(-2) at a conductance (G(te)) of 44+/-5 mS cm(-2). Substitution of either chloride (by nitrate) or sodium (by choline) on both sides of split gill lamellae significantly reduced I(sc) (by 70-80%) and G(te) (by 30-50%). External CsCl (but not BaCl(2) or furosemide) inhibited the negative I(sc) without affecting G(te). Addition of ouabain, BaCl(2) or diphenylamine-2-carboxylate to the internal bath inhibited I(sc) at unchanged G(te). Internal acetazolamide did not affect I(sc) or G(te) across split gill lamellae. Unidirectional Na(+) influx across isolated and perfused posterior gills, however, was reduced by internal acetazolamide by approximately 20% at constant V(te). The results suggest that posterior gills of hyperosmoregulating C. granulatus display a high conductance epithelium that actively absorbs NaCl in a coupled way by an electrogenic mechanism similar to that seen in the thick ascending limb of Henle's loop and, to a minor degree, by an electroneutral mechanism, presumably via apical Na(+)/H(+)- and Cl(-)/HCO(3)( )-antiports. PMID- 12582145 TI - Echoes of bat-pollinated bell-shaped flowers: conspicuous for nectar-feeding bats? AB - Nectar-feeding glossophagine bats searching for flowers are guided by their echolocation system as well as olfactory cues in detecting and recognizing nectar sources. Therefore, chiropterophilous plants, which depend on these bats as pollinators, may be expected to have evolved acoustically conspicuous flowers that facilitate detection. As it is poorly understood how bats manage to find and recognize flowers acoustically, we investigated the echoes of some of the flowers pollinated by bats. Echoes of bell-shaped bat-pollinated flowers have characteristic features with respect to the echoes they reflect to a calling bat and differ from the echoes of leaves or other objects in their surroundings: the echoes are comparatively long and of complex spectral composition. Owing to the specific shape of the flowers, characteristic 'spectral directional patterns' result when the spectra of the echoes are plotted against the angle of sound incidence. We suggest that bats are able to recognize such flowers - and probably other objects as well - not only by a characteristic spectral composition of the echo but also by comparing sequential echoes, at the same time taking into account their exact calling position relative to the object. PMID- 12582146 TI - Jumping and kicking in bush crickets. AB - Bush crickets have long, thin hind legs but jump and kick rapidly. The mechanisms underlying these fast movements were analysed by correlating the activity of femoral muscles in a hind leg with the movements of the legs and body captured in high-speed images. A female Pholidoptera griseoaptera weighing 600 mg can jump a horizontal distance of 300 mm from a takeoff angle of 34 degrees and at a velocity of 2.1 m s(-1), gaining 1350 microJ of kinetic energy. The body is accelerated at up to 114 m s(-2), and the tibiae of the hind legs extend fully within 30 ms at maximal rotational velocities of 13500 deg. s(-1). Such performance requires a minimal power output of 40 mW. Ruddering movements of the hind legs may contribute to the stability of the body once the insect is airborne. During kicking, a hind tibia is extended completely within 10 ms with rotational velocities three times higher at 41800 deg. s(-1). Before a kick, high speed images show no distortions of the hind femoro-tibial joints or of the small semi-lunar groove in the distal femur. Both kicks and jumps can be generated without full flexion of the hind tibiae. Some kicks involve a brief, 40-90 ms, period of co-contraction between the extensor and flexor tibiae muscles, but others can be generated by contraction of the extensor without a preceding co contraction with the flexor. In the latter kicks, the initial flexion of the tibia is generated by a burst of flexor spikes, which then stop before spikes occur in the fast extensor tibiae (FETi) motor neuron. The rapid extension of the tibia can follow directly upon these spikes or can be delayed by as long as 40 ms. The velocity of tibial movement is positively correlated with the number of FETi spikes. The hind legs are 1.5 times longer than the body and more than four times longer than the front legs. The mechanical advantage of the hind leg flexor muscle over the extensor is greater at flexed joint angles and is enhanced by a pad of tissue on its tendon that slides over a protuberance in the ventral wall of the distal femur. The balance of forces in the extensor and flexor muscles, coupled with their changing lever ratio at different joint positions, appears to determine the point of tibial release and to enable rapid movements without an obligatory co-contraction of the two muscles. PMID- 12582147 TI - Acutely altered hemodynamics following venous obstruction in the early chick embryo. AB - In the venous clip model specific cardiac malformations are induced in the chick embryo by obstructing the right lateral vitelline vein with a microclip. Clipping alters venous return and intracardiac laminar blood flow patterns, with secondary effects on the mechanical load of the embryonic myocardium. We investigated the instantaneous effects of clipping the right lateral vitelline vein on hemodynamics in the stage-17 chick embryo. 32 chick embryos HH 17 were subdivided into venous clipped (N=16) and matched control embryos (N=16). Dorsal aortic blood flow velocity was measured with a 20 MHz pulsed Doppler meter. A time series of eight successive measurements per embryo was made starting just before clipping and ending 5h after clipping. Heart rate, peak systolic velocity, time averaged velocity, peak blood flow, mean blood flow, peak acceleration and stroke volume were determined. All hemodynamic parameters decreased acutely after venous clipping and only three out of seven parameters (heart rate, time-averaged velocity and mean blood flow) showed a recovery to baseline values during the 5h study period. We conclude that the experimental alteration of venous return has major acute effects on hemodynamics in the chick embryo. These effects may be responsible for the observed cardiac malformations after clipping. PMID- 12582148 TI - The Karman gait: novel body kinematics of rainbow trout swimming in a vortex street. AB - Most fishes commonly experience unsteady flows and hydrodynamic perturbations during their lifetime. In this study, we provide evidence that rainbow trout Oncorhynchus mykiss voluntarily alter their body kinematics when interacting with vortices present in the environment that are not self-generated. To demonstrate this, we measured axial swimming kinematics in response to changes in known hydrodynamic wake characteristics. We compared trout swimming in the Karman street behind different diameter cylinders (2.5 and 5 cm) at two flow speeds (2.5 and 4.5 L s(-1), where L is total body length) to trout swimming in the free stream and in the cylinder bow wake. Trout swimming behind cylinders adopt a distinctive, previously undescribed pattern of movement in order to hold station, which we term the Karman gait. During this gait, body amplitudes and curvatures are much larger than those of trout swimming at an equivalent flow velocity in the absence of a cylinder. Tail-beat frequency is not only lower than might be expected for a trout swimming in the reduced flow behind a cylinder, but also matches the vortex shedding frequency of the cylinder. Therefore, in addition to choosing to be in the slower flow velocity offered behind a cylinder (drafting), trout are also altering their body kinematics to synchronize with the shed vortices (tuning), using a mechanism that may not involve propulsive locomotion. This behavior is most distinctive when cylinder diameter is large relative to fish length. While tuning, trout have a longer body wavelength than the prescribed wake wavelength, indicating that only certain regions of the body may need to be oriented in a consistent manner to the oncoming vortices. Our results suggest that fish can capture energy from vortices generated by the environment to maintain station in downstream flow. Interestingly, trout swimming in front of a cylinder display lower tail-beat amplitudes and body wave speeds than trout subjected to any of the other treatments, implying that the bow wake may be the most energetically favorable region for a fish to hold station near a cylinder. PMID- 12582149 TI - Skiing across the Greenland icecap: divergent effects on limb muscle adaptations and substrate oxidation. AB - This study investigates the adaptive response of the lower limb muscles and substrate oxidation during submaximal arm or leg exercise after a crossing of the Greenland icecap on cross-country skies. Before and after the 42-day expedition, four male subjects performed cycle ergometer and arm-cranking exercise on two separate days. On each occasion, the subjects exercised at two submaximal loads (arm exercise, 45 W and 100 W; leg exercise, 100 W and 200 W). In addition, peak oxygen uptake ((VO(2max))) was determined for both leg and arm exercise. Before and after the crossing, a muscle biopsy was obtained from the vastus lateralis and the triceps brachii muscles prior to exercise (N=3). After the crossing, body mass decreased by 5.7+/-0.5 kg (in four of four subjects), whereas (VO(2max)) was unchanged in the arm (3.1+/-0.2 l min(-1)) and leg (4.0+/-0.1 l min(-1)). Before the crossing, respiratory exchange ratio (RER) values were 0.84+/-0.02 and 0.96+/ 0.02 during submaximal arm exercise and 0.82+/-0.02 and 0.91+/-0.01 during submaximal leg exercise at the low and high workloads, respectively. After the crossing, RER was lower (in three of four subjects) during arm exercise (0.74+/ 0.02 and 0.81+/-0.01) but was higher (in three of four subjects) during leg exercise (0.92+/-0.02 and 0.96+/-0.01) at the low and high workloads, respectively. Citrate synthase and beta-hydroxy-acyl-CoA-dehydrogenase activity was decreased by approximately 29% in vastus lateralis muscle and was unchanged in triceps brachii muscle. Fat oxidation during submaximal arm exercise was enhanced without a concomitant increase in the oxidative capacity of the triceps brachii muscle after the crossing. This contrasted with decreased fat oxidation during leg exercise, which occurred parallel to a decreased oxidative capacity in vastus lateralis muscle. Although the number of subjects is limited, these results imply that the adaptation pattern after long-term, prolonged, low intensity, whole body exercise may vary dramatically among muscles. PMID- 12582150 TI - Leg morphology and locomotion in birds: requirements for force and speed during ankle flexion. AB - Muscle force production and speed of movement of a bone are not only highly dependent on muscle properties but also on the biomechanical arrangements of the musculoskeletal systems. The aim of this study was to investigate whether the leverages of a leg flexion system alone could be used to trace adaptations to different locomotion patterns by different groups of birds. We focused on ankle flexion, and measured the length (tmt) of the tarsometatarsus, representing the out-force lever arm, and the distance (d) between the ankle joint and the tendon insertion of the flexor muscle of the tarsometatarsus, representing the in-force lever arm. By the use of residuals from regressions, tmt and d were made independent of body mass, and d independent of tmt, forming indices of the lever arms, d(index) and tmt(index). The investigation included 67 bird species divided into six groups according to differences in their hind limb movements and requirements of force and speed. These were birds that walk/run/hop (WH), climb (C) or hang (H), birds of prey (BOP), fast swimmers (FS) and slow swimmers (SS). Predictions for each group correlating their requirements for force and speed are made, based on biomechanical and ecological factors, and the lengths of the moment arms are calculated. The results show that the means for the groups could largely be separated from the norm (i.e. zero), and in many cases the predictions are fulfilled. d is significantly larger than average in species affected by strong forces, for example, gravity (BOP and C), but shorter in species affected only by drag (WH, FS and SS). No differences associated with drag due to differences in medium density were seen. Furthermore, the tarsometatarsus is longer than average only in the BOP species, and shorter in the SS species. Discriminant analysis reveals that using our predictions there is a 53.7% chance of placing a species in the correct group, compared with the 17% chance expected if the species are randomly placed in a group. PMID- 12582152 TI - Oligomerization of the human ARF tumor suppressor and its response to oxidative stress. AB - The tumor suppressor ARF plays an important role as an inhibitor of the Mdm2 mediated degradation of p53. Here we demonstrate that human ARF (p14ARF) can form homo-oligomers. The stability of the oligomers is favored by oxidizing agents in a reversible fashion and involves all three cysteine residues in p14ARF. Furthermore, the effect of p14ARF in clonogenic assays is moderately but reproducibly increased by the mutation of its cysteine residues. We also observed that altering the amino terminus of p14ARF resulted in the appearance of remarkably stable oligomers. This indicates that the amino terminus of p14ARF interferes with the ability of the protein to form multimeric complexes. These observations suggest that p14ARF activity may be linked to its oligomerization status and sensitive to the redox status of the cell. PMID- 12582153 TI - c-MIR, a human E3 ubiquitin ligase, is a functional homolog of herpesvirus proteins MIR1 and MIR2 and has similar activity. AB - Kaposi's sarcoma associated-herpes virus encodes two proteins, MIR (modulator of immune recognition) 1 and 2, which are involved in the evasion of host immunity. MIR1 and 2 have been shown to function as an E3 ubiquitin ligase for immune recognition-related molecules (e.g. major histocompatibility complex class I, B7 2, and ICAM-1) through the BKS (bovine herpesvirus 4, Kaposi's sarcoma associated herpes virus, and Swinepox virus) subclass of plant homeodomain (PHD) domain, termed the BKS-PHD domain. Here we show that the human genome also encodes a novel BKS-PHD domain-containing protein that functions as an E3 ubiquitin ligase and whose putative substrate is the B7-2 co-stimulatory molecule. This novel E3 ubiquitin ligase was designated as c-MIR (cellular MIR) based on its functional and structural similarity to MIR1 and 2. Forced expression of c-MIR induced specific down-regulation of B7-2 surface expression through ubiquitination, rapid endocytosis, and lysosomal degradation of the target molecule. This specific targeting was dependent upon the binding of c-MIR to B7-2. Replacing the BKS-PHD domain of MIR1 with the corresponding domain of c-MIR did not alter MIR1 function. The discovery of c-MIR, a novel E3 ubiquitin ligase, highlights the possibility that viral immune regulatory proteins originated in the host genome and presents unique functions of BKS-PHD domain-containing proteins in mammals. PMID- 12582154 TI - Assembly and secretion of very low density lipoproteins containing apolipoprotein B48 in transfected McA-RH7777 cells. Lack of evidence that palmitoylation of apolipoprotein B48 is required for lipoprotein secretion. AB - We examined the role of S-linked palmitoylation of human apolipoprotein (apo) B in the assembly and secretion of very low density lipoproteins using recombinant human apoB48. There are four free cysteine residues (Cys(1085), Cys(1396), Cys(1478), and Cys(1635)) within apoB48 that potentially can be palmitoylated. All four cysteine residues were substituted with serine by site-specific mutagenesis. The mutant protein was expressed in transfected rat hepatoma McA RH7777 cells. Metabolic labeling of the stably transfected cells with iodopalmitic acid analog showed that the mutant apoB48 lacked palmitoylation. The lack of palmitoylation had little impact on the ability of apoB48 to assemble and secrete very low density lipoproteins or high density lipoproteins. Immunocytochemistry experiments using confocal microscopy failed to reveal any major alterations in the intracellular distribution of the mutant apoB48 at steady state. Pulse-chase analysis combined with subcellular fractionation showed no apparent deficiency in the movement of the mutant apoB48 protein from the endoplasmic reticulum to cis/medial Golgi. However, the mutant apoB48 lacking palmitoylation showed retarded movement toward the distal Golgi and increased association (>2-fold) with the membranes of the secretory compartments. A marginal decrease (by 15-20%) in secretion efficiency as compared with that of wild type apoB48 was also observed. These results suggest that lack of palmitoylation may influence the partitioning of apoB48 between microsomal membranes and microsomal lumen, but it does not compromise the ability of apoB48 to assemble lipoproteins. PMID- 12582155 TI - Matrix assembly induction and cell migration and invasion inhibition by a 13 amino acid fibronectin peptide. AB - Fibronectin (FN) is an extracellular matrix (ECM) protein involved in tumor growth and metastasis. Five human FN cDNA segments encoding for FN fragments, all starting with the II1 repeat and ending with different C-terminal extensions, have been stably expressed in chick embryo fibroblasts (CEF). These FN cDNAs induce the formation of an organized ECM in CEF as long as they retain a sequence coding for a 13-amino acid stretch (FN13), with collagen binding activity, localized between type II2 and I7 repeats. An FN13 synthetic peptide induces in control CEF the assembly of an FN-ECM comparable with that observed in CEF expressing FN fragments. The activity of FN13 is specific for its amino acid sequence, although the cysteine present in the 6th position can be substituted with a polar serine without affecting the induction of a fibrillar FN-ECM. A less fibrillar matrix is induced by FN13-modified peptides in which the cysteine is methylated or substituted by a non-polar alanine. FN13 induces the assembly of an FN-ECM also in Rous sarcoma virus-transformed CEF lacking the ECM and in hepatoma (SK-Hep1) and fibrosarcoma (HT-1080) human cell lines. FN13 also promotes the adhesion of CEF and Rous sarcoma virus-CEF at levels comparable with those obtained with purified intact FN. Finally, FN13 inhibits the migratory and invasive properties of tumorigenic cells, whereas intact FN favors their migration. All FN13-modified peptides show similar effects, although with reduced efficiency. None of these activities is supported by a scrambled peptide. These data suggest a possible role of FN13 in tumor growth and metastasis inhibition and its possible use as anti-tumorigenic agent. PMID- 12582156 TI - Gene array and protein expression profiles suggest post-transcriptional regulation during CD8+ T cell differentiation. AB - Peripheral CD8(+) T cells circulate in a quiescent naive state until they are primed by specific antigen and differentiate into effector cells. In the effector state, CD8(+) T cells acquire cytolytic activity and produce increased levels of cytokines such as interferon-gamma. They also exhibit increased T cell receptor sensitivity, decreased CD28 dependence, and become inhibitable by CTLA-4 and other negative regulatory pathways. We hypothesized that one mechanism by which these two states are regulated is via differential expression of specific genes. To this end, basal gene expression profiles of naive and effector 2C TCR transgenic x RAG2(-/-) CD8(+) T cells were analyzed using Affymetrix arrays representing 11,000 genes. Of the 177 differentially expressed known genes, 68 were expressed at higher levels in effector cells, but 109 were more abundant in naive cells, supporting the notion that the naive state is not passive. Expression of genes related to metabolism, actin cytoskeletal dynamics, and effector function increased with priming, whereas expression of putative anti proliferative genes decreased. Semiquantitative reverse transcription-PCR was utilized as a secondary validation for selected transcripts, and Western blot analysis was used to examine protein expression for molecules of interest. Surprisingly, for 24 genes examined, 12 showed discordant protein versus mRNA expression. In summary, our study indicates that: 1) not only does the expression of some genes in naive CD8(+) T cells become up-regulated upon priming, but the expression of other genes is down-regulated as well and 2) the complexities of T cell differentiation include regulation at the post-transcriptional level. PMID- 12582157 TI - An essential function of tapasin in quality control of HLA-G molecules. AB - Tapasin plays an important role in the quality control of major histocompatibility complex (MHC) class I assembly, but its precise function in this process remains controversial. Whether tapasin participates in the assembly of HLA-G has not been studied. HLA-G, an MHC class Ib molecule that binds a more restricted set of peptides than class Ia molecules, is a particularly interesting molecule, because during assembly, it recycles between the endoplasmic reticulum (ER) and the cis-Golgi until it is loaded with a high affinity peptide. We have taken advantage of this unusual trafficking property of HLA-G and its requirement for high affinity peptides to demonstrate that a critical function of tapasin is to transform class I molecules into a high affinity, peptide-receptive form. In the absence of tapasin, HLA-G molecules cannot bind high affinity peptides, and an abundant supply of peptides cannot overcome the tapasin requirement for high affinity peptide loading. The addition of tapasin renders HLA-G molecules capable of loading high affinity peptides and of transporting to the surface, suggesting that tapasin is a prerequisite for the binding of high-affinity ligands. Interestingly, the "tapasin-dependent" HLA-G molecules are not empty in the absence of tapasin but are in fact associated with suboptimal peptides and continue to recycle between the ER and the cis-Golgi. Together with the finding that empty HLA-G heterodimers are strictly retained in the ER and degraded, our data suggest that MHC class I molecules bind any available peptides to avoid ER mediated degradation and that the peptides are in turn replaced by higher affinity peptides with the aid of tapasin. PMID- 12582158 TI - Loss of murine Na+/myo-inositol cotransporter leads to brain myo-inositol depletion and central apnea. AB - myo-Inositol (Ins) and its polyphosphoinositide derivatives that are important in membrane signaling have long been held to play a special role in brain metabolism. As polyphosphoinositides turn over rapidly and are exceptionally abundant in nervous tissue, high Ins levels in the range of 2-15 mm that have been observed in brain may be necessary to maintain the rates of phosphoinositide synthesis in diverse membrane locations within neurons. Cellular concentration gradients of this magnitude indicate a dependence on active Ins transport, especially at the time of growth and differentiation. The Na(+)/myo-inositol cotransporter (SMIT1 or SLC5A3) gene is highly expressed prenatally in the central nervous system and placenta. To gain more insight into brain Ins metabolism, while ascertaining the importance of SMIT1 as a transporter, we generated mice with a homozygous targeted deletion of this gene. Newborn SMIT1(-/ ) animals have no evidence of SMIT1 mRNA, a 92% reduction in the level of brain Ins, an 84% reduction in whole body Ins, and expire shortly after birth due to hypoventilation. Gross pathologic and light microscopic examinations of each organ, as well as the placenta, of embryonic day 18.5 fetuses at near term gestation were normal. Based on [(3)H]acetate incorporation into phospholipids of lung tissue explants, immunostaining of lung tissue for surfactant protein A, B, and C, and electron microscopic examination of alveolar cells, there was no evidence of abnormal pulmonary surfactant production by type 2 pneumocytes in lung. Although no histologic lesions were detected in the nervous system, electrophysiological studies of the brainstem pre-Botzinger respiratory control center demonstrated an abnormal rhythm discharge with periods of central apnea. The cause of death can be explained by the regulatory defect in brainstem control of ventilation. This model demonstrates the critical importance of SMIT1 in the developing nervous system. The high affinity SMIT1 transporter is responsible for the Ins concentration gradient in the murine fetal-placental unit. PMID- 12582159 TI - A new beta-catenin-dependent activation domain in T cell factor. AB - Transcription of the lymphoid enhancer factor-1 (LEF1) gene is aberrantly activated in sporadic colon cancer, whereas this gene is not expressed in the normal adult colon. We have shown previously that promoter 1 of the LEF1 gene is activated by T cell factor (TCF)-beta-catenin complexes in transient transfection assays, suggesting that LEF1 is a target of the Wnt pathway in colon cancer. To further explore the link between LEF1 expression and the Wnt pathway, we studied two response elements in the promoter. Surprisingly we found that the LEF1 promoter is selectively activated by specific isoforms of the LEF/TCF transcription factor family that contain an alternative C-terminal "E" tail. These isoforms, TCF-1E and TCF-4E, activate the promoter in a beta-catenin dependent manner. We show that a complete E-tail domain is necessary for full activity and delimits residues within two highly conserved peptide motifs within the tail that are required (KKCRARFG; WCXXCRRKKKC). These peptide motifs are not only conserved among the TCF family members but are also found in two newly identified DNA-binding proteins named papillomavirus binding factor and GLUT4 enhancer factor. This study thus identifies a new and unique set of motifs used by the Wnt pathway for target gene regulation. PMID- 12582160 TI - Decorin binds fibrinogen in a Zn2+-dependent interaction. AB - We have previously shown that decorin, a member of the small leucine-rich proteoglycan family of extracellular matrix proteoglycans/glycoproteins is a Zn(2+) metalloprotein at physiological Zn(2+) concentrations (Yang, V. W-C., LaBrenz, S. R., Rosenberg, L. C., McQuillan, D., and Hook, M. (1999) J. Biol. Chem. 274, 12454-12460). We now report that the decorin proteoglycan binds fibrinogen in the presence of Zn(2+). The fibrinogen-binding site is located in the N-terminal domain of the decorin core protein and a 45-amino acid peptide representing this domain binds to the fibrinogen D fragment with an apparent K(D) of 1.7 x 10(-6) m, as determined from fluorescence polarization data. Furthermore, we show that Zn(2+) promotes the self-association of decorin. The N terminal domain of the core protein also mediates this activity. The results of solid-phase binding assays and gel filtration chromatography suggest that the N terminal domain of decorin, when present at low micromolar concentrations, forms an oligomer in a Zn(2+)-dependent manner. Thus, Zn(2+) appears to play a pivotal role in the interactions and biological function of decorin. PMID- 12582161 TI - The UDP-glucuronosyltransferase 1A9 enzyme is a peroxisome proliferator-activated receptor alpha and gamma target gene. AB - Peroxisome proliferator-activated receptor (PPAR) alpha and gamma are ligand activated transcription factors belonging to the nuclear receptor family. PPAR alpha mediates the hypolipidemic action of the fibrates, whereas PPAR gamma is a receptor for the antidiabetic glitazones. In the present study, the UDP glucuronosyltransferase (UGT) 1A9 enzyme is identified as a PPAR alpha and PPAR gamma target gene. UGTs catalyze the glucuronidation reaction, which is a major pathway in the catabolism and elimination of numerous endo- and xenobiotics. Among the UGT1A family enzymes, UGT1A9 metabolizes endogenous compounds, including catecholestrogens, and xenobiotics, such as fibrates and to a lesser extent troglitazone. Treatment of human hepatocytes and macrophages and murine adipocytes with activators of PPAR alpha or PPAR gamma resulted in an enhanced UGT1A9 expression and activity. In addition, disruption of the PPAR alpha gene in mice completely abolished the PPAR alpha agonist-induced UGT1A9 mRNA and activity levels. A PPAR response element was identified in the promoter of UGT1A9 at positions -719 to -706 bp by transient transfection and electromobility shift assays. Considering the role of UGT1A9 in catecholestrogen metabolism, PPAR alpha and PPAR gamma activation may contribute to the protection against genotoxic catecholestrogens by stimulating their inactivation in glucuronide derivatives. Furthermore, since UGT1A9 is involved in the catabolism of fibrates, these results suggest that PPAR alpha and PPAR gamma may control the intracellular level of active fibrates. PMID- 12582162 TI - The p38 and MK2 kinase cascade phosphorylates tuberin, the tuberous sclerosis 2 gene product, and enhances its interaction with 14-3-3. AB - Tuberous sclerosis complex (TSC) is a genetic disease caused by mutations in either TSC1 or TSC2 tumor suppressor genes. TSC1 and TSC2 (also known as hamartin and tuberin, respectively) form a functional complex and negatively regulate cell growth by inhibiting protein synthesis. 14-3-3 binds to TSC2 and may inhibit TSC2 function. We have reported previously that phosphorylation of serine 1210 (Ser(1210)) in TSC2 is essential for 14-3-3 binding. Here we show that serum and anisomycin enhance the interaction between TSC2 and 14-3-3 by stimulating phosphorylation of Ser(1210). Activation of p38 MAP kinase (p38) is essential for the stimulating effect of serum and anisomycin although p38 is not directly responsible for the phosphorylation of Ser(1210) in TSC2. Both in vitro and in vivo experiments demonstrate that the p38-activated kinase MK2 (also known as MAPKAPK2) is directly responsible for the phosphorylation of Ser(1210). Our data show that anisomycin stimulates phosphorylation of Ser(1210) of TSC2 via the p38 MK2 kinase cascade. Phosphorylation of TSC2 by MK2 creates a 14-3-3 binding site and thus regulates the cellular function of the TSC2 tumor suppressor protein. PMID- 12582164 TI - In search of the energetic role of peptide hydrogen bonds. PMID- 12582163 TI - A single-nucleotide deletion leads to rapid degradation of TAP-1 mRNA in a melanoma cell line. AB - Both viruses and tumors evade cytotoxic T lymphocyte-mediated host immunity by down-regulation of antigen presentation machineries. This can be achieved by either down-regulation of transcription of antigen presentation genes or posttranslational inactivation of proteins involved in antigen presentation. In this study, a major histocompatibility complex (MHC) class I-deficient melanoma cell line, SK-MEL-19, was found deficient in the expression of the transporter associated with antigen processing (TAP)-1 mRNA even after IFN-gamma stimulation, despite its active transcription of the TAP-1 gene. This abnormality was caused by a single-nucleotide deletion at position +1489 of the TAP-1 gene and was corrected by cycloheximide, which inhibits RNA degradation. Using an inducible Tet-Off system, we demonstrated that deletion of the nucleotide resulted in a >2 fold decrease in the half-life of TAP-1 mRNA. However, the decrease of the half life of TAP-1 mRNA is not mediated by nonsense-mediated mRNA decay because deletions of two additional nucleotides in the region, which corrected the nonsense mutation, did not restore TAP-1 mRNA stability. To our knowledge, this is the first evidence that the degradation of mRNA of an antigen presentation gene is involved in HLA class I down-regulation in malignant cells. PMID- 12582165 TI - Development of an efficient "substrate-trapping" mutant of Src homology phosphotyrosine phosphatase 2 and identification of the epidermal growth factor receptor, Gab1, and three other proteins as target substrates. AB - Src homology containing phosphotyrosine phosphatase 2 (SHP2) is a positive effector of growth factor, cytokine, and integrin signaling. However, neither its physiological substrate nor its mechanism of action in tyrosine kinase signaling has been demonstrated. We reasoned that the identification of physiological substrates of SHP2 would be a stepping stone in elucidating its mechanism of action, and, thus, we constructed a potent trapping mutant of SHP2. Surprisingly, the frequently used Asp to Ala substitution did not give rise to a trapping mutant. However, we were able to develop an efficient trapping mutant of SHP2 by introducing Asp to Ala and Cys to Ser double mutations. The double mutant (DM) protein identified the epidermal growth factor receptor (EGFR), the Grb2 binder 1, and three other, as yet unidentified, phosphotyrosyl proteins as candidate physiological substrates. Given that substrate trapping occurred in intact cells and that the interaction was very specific, it is highly likely that EGFR and Gab1 represent physiological SHP2 substrates. Therefore, the DM protein would serve as an important tool in future SHP2 studies, including identification of p190, p150, and p90. PMID- 12582166 TI - Oncogenic potential of a dominant negative mutant of interferon regulatory factor 3. AB - Interferon regulatory factor 3 (IRF3) is activated in response to various environmental stresses including viral infection and DNA-damaging agents. However, the biological function of IRF3 in cell growth is not well understood. We demonstrated that IRF3 markedly inhibited growth and colony formation of cells. IRF3 blocked DNA synthesis and induced apoptosis. Based on this negative control of cell growth by IRF3, we examined whether functional loss of IRF3 may contribute to oncogenic transformation. IRF3 activity was specifically inhibited by expression of its dominant negative mutant. This mutant lacks a portion of the DNA binding domain like IRF3a, an alternative splice form of IRF3 in the cells. This dominant negative inhibition blocked expression of specific IRF3 target genes. Mutant IRF3 efficiently transformed NIH3T3 cells, as demonstrated by anchorage-independent growth in soft agar and tumorigenicity in nude mice. These results imply that IRF3 may function as a tumor suppressor and suggest a possible role for the relative levels of IRF3 and its dominant negative mutant in tumorigenesis. PMID- 12582167 TI - Ordered ATP hydrolysis in the gamma complex clamp loader AAA+ machine. AB - The gamma complex couples ATP hydrolysis to the loading of beta sliding clamps onto DNA for processive replication. The gamma complex structure shows that the clamp loader subunits are arranged as a circular heteropentamer. The three gamma motor subunits bind ATP, the delta wrench opens the beta ring, and the delta' stator modulates the delta-beta interaction. Neither delta nor delta' bind ATP. This report demonstrates that the delta' stator contributes a catalytic arginine for hydrolysis of ATP bound to the adjacent gamma(1) subunit. Thus, the delta' stator contributes to the motor function of the gamma trimer. Mutation of arginine 169 of gamma, which removes the catalytic arginines from only the gamma(2) and gamma(3) ATP sites, abolishes ATPase activity even though ATP site 1 is intact and all three sites are filled. This result implies that hydrolysis of the three ATP molecules occurs in a particular order, the reverse of ATP binding, where ATP in site 1 is not hydrolyzed until ATP in sites 2 and/or 3 is hydrolyzed. Implications of these results to clamp loaders of other systems are discussed. PMID- 12582168 TI - A new class of N-hydroxycinnamoyltransferases. Purification, cloning, and expression of a barley agmatine coumaroyltransferase (EC 2.3.1.64). AB - Agmatine coumaroyltransferase (ACT), which catalyzes the first step in the biosynthesis of antifungal hydroxycinnamoylagmatine derivatives, was purified to apparent homogeneity from 3-day-old etiolated barley (Hordeum vulgare L.) seedlings. The enzyme was highly specific for agmatine as acyl acceptor and had the highest specificity for p-coumaroyl-CoA among various acyl donors with a specific activity of 29.7 nanokatal x mg(-1) protein. Barley ACT was found to be a single polypeptide chain of 48 kDa with a pI of 5.20 as determined by isoelectric focusing. The 15 N-terminal amino acid residues were identified by micro-sequencing of the native protein and were used to clone a full-length barley ACT cDNA that predicted a protein of 439 amino acid residues. The sequence was devoid of N-terminal signal peptide, suggesting a cytosolic localization of barley ACT. Recombinant ACT produced and affinity-purified from Escherichia coli had a specific activity of 189 nanokatal x mg(-1) protein, thus confirming the identity of the purified native protein. A partial cDNA sequence for ACT was obtained from wheat that predicted a protein of 353 amino acid residues and had 95% sequence identity to barley ACT. Two motifs in the amino acid sequence reveal that barley ACT represents a new class of N-hydroxycinnamoyltransferases belonging to the transferase superfamily. The barley ACT is unique in producing the precursor of hordatine, a proven antifungal factor that may be directed toward Blumeria graminis. PMID- 12582169 TI - Identification of a surface charged residue in the S3-S4 linker of the pacemaker (HCN) channel that influences activation gating. AB - I(f), encoded by the hyperpolarization-activated cyclic nucleotide-modulated (HCN) channel family, is a key player in cardiac and neuronal pacing. Although HCN channels structurally resemble voltage-gated K(+) (Kv) channels, their structure-function correlation is much less clear. Here we probed the functional importance of the HCN1 S3-S4 linker by multiple substitutions of its residues. Neutralizing Glu(235), an acidic S3-S4 linker residue conserved in all hyperpolarization-activated channels, by Ala substitution produced a depolarizing activation shift (V(12) = -65.0 +/- 0.7 versus -70.6 +/- 0.7 mV for wild-type HCN1); the charge-reversed mutation E235R shifted activation even more positively (-56.2 +/- 0.5 mV). Increasing external Mg(2+) mimicked the progressive rightward shifts of E235A and E235R by gradually shifting activation (V(12) = 1 < 3 < 10 < 30 mm); Delta V(12) induced by 30 mm Mg(2+) was significantly attenuated for E235A (+7.9 +/- 1.2 versus +11.3 +/- 0.9 mV for wild-type HCN1) and E235R (+3.3 +/- 1.4 mV) channels, as if surface charges were already shielded. Consistent with an electrostatic role, the energetic changes associated with Delta V(12) resulting from various Glu(235) substitutions (i.e. Asp, Ala, Pro, His, Lys, and Arg) displayed a strong correlation with their charges (Delta Delta G = -2.1 +/- 0.3 kcal/mol/charge; r = 0.94). In contrast, D233E, D233A, D233G, and D233R did not alter activation gating. D233C (in C318S background) was also not externally accessible when probed with methanethiosulfonate ethylammonium (MTSEA). We conclude that the S3-S4 linker residue Glu(235) influences activation gating, probably by acting as a surface charge. PMID- 12582170 TI - Redox-regulated rotational coupling of receptor protein-tyrosine phosphatase alpha dimers. AB - Receptor protein-tyrosine phosphatase alpha (RPTP alpha) constitutively forms dimers in the membrane, and activity studies with forced dimer mutants of RPTP alpha revealed that rotational coupling of the dimer defines its activity. The hemagglutinin (HA) tag of wild type RPTP alpha and of constitutively dimeric, active RPTP alpha-F135C with a disulfide bond in the extracellular domain was not accessible for antibody, whereas the HA tag of constitutively dimeric, inactive RPTP alpha-P137C was. All three proteins were expressed on the plasma membrane to a similar extent, and the accessibility of their extracellular domains did not differ as determined by biotinylation studies. Dimerization was required for masking the HA tag, and we identified a region in the N terminus of RPTP alpha that was essential for the effect. Oxidative stress has been shown to induce a conformational change of the membrane distal PTP domain (RPTP alpha-D2). Here we report that H(2)O(2) treatment of cells induced a change in rotational coupling in RPTP alpha dimers as detected using accessibility of an HA tag in the extracellular domain as a read-out. The catalytic site Cys(723) in RPTP alpha-D2, which was required for the conformational change of RPTP alpha-D2 upon H(2)O(2) treatment, was essential for the H(2)O(2)-induced increase in accessibility. These results show for the first time that a conformational change in the intracellular domain of RPTP alpha led to a change in conformation of the extracellular domains, indicating that RPTPs have the capacity for inside-out signaling. PMID- 12582171 TI - A specific domain of Gialpha required for the transactivation of Gialpha by tubulin is implicated in the organization of cellular microtubules. AB - G(s)alpha, G(i)alpha(1), and G(q)alpha subunits bind tubulin with high affinity, whereas transducin (G(t)alpha) does not. The interaction between tubulin and Galpha, which also involves the direct transfer of GTP from tubulin to Galpha (transactivation), is not yet fully understood. This study, using chimeras of G(i)alpha and G(t)alpha, showed that the G(i)alpha (215-295) segment converted G(t)alpha to bind to tubulin and this chimera (chimera 1) could be transactivated by tubulin. Insertion of G(t)alpha (237-270) into chimera 1 to form chimera 2 resulted in a protein that, like G(t)alpha, did not bind tubulin. Thus, it was thought that the G(i)alpha (237-270) domain was essential to modulate the binding of G(i)alpha(1) to tubulin. Surprisingly, when domain (237-270) of G(i)alpha was replaced by G(t)alpha (237-270) to form chimera 3, the chimera bound to tubulin with a similar affinity (K(D) congruent with 120 nm) as wild-type G(i)alpha(1). However, even though chimera 3 displayed normal GTP binding, it was not transactivated by GTP-tubulin. Furthermore, when these chimeras were expressed in COS-1 cells, cellular processes in cells overexpressing G(i)alpha(1) or chimera 1 were more abundant and longer than those in native cells. Galpha was seen throughout the length of the process. Morphology of cells expressing chimera 2 was identical to controls. Consistent with the role of Chimera 3 as a "dominant negative" Galpha, cells transfected with chimera 3 had only few truncated processes. This study demonstrates that although G(i)alpha (237-270) is not obligatory for the binding of G(i)alpha to tubulin, it is crucial for the transactivation of Galpha by tubulin. These results also suggest that the transactivation of Galpha by tubulin may play an important role in modulating microtubule organization and cell morphology. PMID- 12582172 TI - A receptor-binding region in Escherichia coli alpha-haemolysin. AB - Escherichia coli alpha-hemolysin (HlyA) is a 107-kDa protein toxin with a wide range of mammalian target cells. Previous work has shown that glycophorin is a specific receptor for HlyA in red blood cells (Cortajarena, A. L., Goni, F. M., and Ostolaza, H. (2001) J. Biol. Chem. 276, 12513-12519). The present study was aimed at identifying the glycophorin-binding region in the toxin. Data in the literature pointed to a short amino acid sequence near the C terminus as a putative receptor-binding domain. Previous sequence analyses of several homologous toxins that belong, like HlyA, to the so-called RTX toxin family revealed a conserved region that corresponded to residues 914-936 of HlyA. We therefore prepared a deletion mutant lacking these residues (HlyA Delta 914-936) and found that its hemolytic activity was decreased by 10,000-fold with respect to the wild type. This deletion mutant was virtually unable to bind human and horse red blood cells or to bind pure glycophorin in an affinity column. The peptide Trp914-Arg936 had no lytic activity of its own, but it could bind glycophorin reconstituted in lipid vesicles. Moreover, the peptide Trp914-Arg936 protected red blood cells from hemolysis induced by wild type HlyA. It was concluded that amino acid residues 914-936 constitute a major receptor-binding region in alpha-hemolysin. PMID- 12582173 TI - Folding pathway mediated by an intramolecular chaperone. A functional peptide chaperone designed using sequence databases. AB - Catalytic domains of several prokaryotic and eukaryotic protease families require dedicated N-terminal propeptide domains or "intramolecular chaperones" to facilitate correct folding. Amino acid sequence analysis of these families establishes three important characteristics: (i) propeptides are almost always less conserved than their cognate catalytic domains, (ii) they contain a large number of charged amino acids, and (iii) propeptides within different protease families display insignificant sequence similarity. The implications of these findings are, however, unclear. In this study, we have used subtilisin as our model to redesign a peptide chaperone using information databases. Our goal was to establish the minimum sequence requirements for a functional subtilisin propeptide, because such information could facilitate subsequent design of tailor made chaperones. A decision-based computer algorithm that maintained conserved residues but varied all non-conserved residues from a multiple protein sequence alignment was developed and utilized to design a novel peptide sequence (ProD). Interestingly, despite a difference of 5 pH units between their isoelectric points and despite displaying only 16% sequence identity with the wild-type propeptide (ProWT), ProD chaperones folding and functions as a potent subtilisin inhibitor. The computed secondary structures and hydrophobic patterns within these two propeptides are similar. However, unlike ProWT, ProD adopts a well defined alpha-beta conformation as an isolated peptide and forms a stoichiometric complex with mature subtilisin. The CD spectra of this complex is similar to ProWT.subtilisin. Our results establish that despite low sequence identity and dramatically different charge distribution, both propeptides adopt similar structural scaffolds. Hence, conserved scaffolds and hydrophobic patterns, but not absolute charge, dictate propeptide function. PMID- 12582174 TI - Crystal structure and biochemical analysis of the MutS.ADP.beryllium fluoride complex suggests a conserved mechanism for ATP interactions in mismatch repair. AB - During mismatch repair ATP binding and hydrolysis activities by the MutS family proteins are important for both mismatch recognition and for transducing mismatch recognition signals to downstream repair factors. Despite intensive efforts, a MutS.ATP.DNA complex has eluded crystallographic analysis. Searching for ATP analogs that strongly bound to Thermus aquaticus (Taq) MutS, we found that ADP.beryllium fluoride (ABF), acted as a strong inhibitor of several MutS family ATPases. Furthermore, ABF promoted the formation of a ternary complex containing the Saccharomyces cerevisiae MSH2.MSH6 and MLH1.PMS1 proteins bound to mismatch DNA but did not promote dissociation of MSH2.MSH6 from mismatch DNA. Crystallographic analysis of the Taq MutS.DNA.ABF complex indicated that although this complex was very similar to that of MutS.DNA.ADP, both ADP.Mg(2+) moieties in the MutS. DNA.ADP structure were replaced by ABF. Furthermore, a disordered region near the ATP-binding pocket in the MutS B subunit became traceable, whereas the equivalent region in the A subunit that interacts with the mismatched nucleotide remained disordered. Finally, the DNA binding domains of MutS together with the mismatched DNA were shifted upon binding of ABF. We hypothesize that the presence of ABF is communicated between the two MutS subunits through the contact between the ordered loop and Domain III in addition to the intra-subunit helical lever arm that links the ATPase and DNA binding domains. PMID- 12582175 TI - Two subunits of glycosylphosphatidylinositol transamidase, GPI8 and PIG-T, form a functionally important intermolecular disulfide bridge. AB - Many eukaryotic proteins are tethered to the plasma membrane via glycosylphosphatidylinositol (GPI). GPI transamidase is localized in the endoplasmic reticulum and mediates post-translational transfer of preformed GPI to proteins bearing a carboxyl-terminal GPI attachment signal. Mammalian GPI transamidase is a multimeric complex consisting of at least five subunits. Here we report that two subunits of mammalian GPI transamidase, GPI8 and PIG-T, form a functionally important disulfide bond between conserved cysteine residues. GPI8 and PIG-T mutants in which relevant cysteines were replaced with serines were unable to fully restore the surface expression of GPI-anchored proteins upon transfection into their respective mutant cells. Microsomal membranes of these transfectants had markedly decreased activities in an in vitro transamidase assay. The formation of this disulfide bond is not essential but required for full transamidase activity. Antibodies against GPI8 and PIG-T revealed that endogenous as well as exogenous proteins formed a disulfide bond. Furthermore trypanosome GPI8 forms a similar intermolecular disulfide bond via its conserved cysteine residue, suggesting that the trypanosome GPI transamidase is also a multimeric complex likely containing the orthologue of PIG-T. We also demonstrate that an inactive human GPI transamidase complex that consists of non-functional GPI8 and four other components was co-purified with the proform of substrate proteins, indicating that these five components are sufficient to hold the substrate proteins. PMID- 12582176 TI - High-throughput immunoblotting. Ubiquitiin-like protein ISG15 modifies key regulators of signal transduction. AB - ISG15 is a ubiquitin-like protein that conjugates to numerous proteins in cells treated with interferon or lipopolysaccharide. Dysregulation of protein ISG15 modification (ISGylation) in mice leads to decreased life expectancy, brain cell injury, and hypersensitivity to interferon. Although ISG15 was identified more than two decades ago, the exact biochemical and physiological functions of ISG15 modification remain unknown, and the proteins targeted by ISG15 have not been identified. The major purpose of this work was to identify ISG15 targets among well characterized proteins that could be used as models for biological studies. We purified ISGylated proteins from human thymus by immunoaffinity chromatography and analyzed ISG15 conjugates by a high-throughput Western blot screen (PowerBlot). We found that three key regulators of signal transduction, phospholipase Cgamma1, Jak1, and ERK1 are modified by ISG15. In addition to that, we demonstrate that transcription factor Stat1, an immediate substrate of Jak1 kinase, is also ISGylated. Using whole cell protein extracts and phospholipase Cgamma1 as an example we demonstrate that ISG15 conjugates are not accumulated in cells treated with specific inhibitors of proteasomes. Our work suggests a role for ISG15 in the regulation of multiple signal transduction pathways and offers attractive models to further elucidate the biochemical function of ISGylation. PMID- 12582177 TI - RNase L mediates transient control of the interferon response through modulation of the double-stranded RNA-dependent protein kinase PKR. AB - The transient control of diverse biological responses that occurs in response to varied forms of stress is often a highly regulated process. During the interferon (IFN) response, translational repression due to phosphorylation of eukaryotic initiation factor 2alpha, eIF2alpha, by the double-stranded RNA-dependent protein kinase, PKR, constitutes a means of inhibiting viral replication. Here we show that the transient nature of the IFN response against acute viral infections is regulated, at least in part, by RNase L. During the IFN antiviral response in RNase L-null cells, PKR mRNA stability was enhanced, PKR induction was increased, and the phosphorylated form of eIF2alpha appeared with extended kinetics compared with similarly treated wild type cells. An enhanced IFN response in RNase L-null cells was also demonstrated by monitoring inhibition of viral protein synthesis. Furthermore, ectopic expression of RNase L from a plasmid vector prevented the IFN induction of PKR. These results suggest a role for RNase L in the transient control of the IFN response and possibly of other cytokine and stress responses. PMID- 12582178 TI - Mapping the collagen-binding site in the von Willebrand factor-A3 domain. AB - The multimeric glycoprotein von Willebrand factor (VWF) mediates platelet adhesion to collagen at sites of vascular damage. The binding site for collagen types I and III is located in the VWF-A3 domain. Recently, we showed that His(1023), located near the edge between the "front" and "bottom" faces of A3, is critical for collagen binding (Romijn, R. A., Bouma, B., Wuyster, W., Gros, P., Kroon, J., Sixma, J. J., and Huizinga, E. G. (2001) J. Biol. Chem. 276, 9985 9991). To map the binding site in detail, we introduced 22 point mutations in the front and bottom faces of A3. The mutants were expressed as multimeric VWF, and binding to collagen type III was evaluated in a solid-state binding assay and by surface plasmon resonance. Mutation of residues Asp(979), Ser(1020), and His(1023) nearly abolished collagen binding, whereas mutation of residues Ile(975), Thr(977), Val(997), and Glu(1001) reduced binding affinity about 10 fold. Together, these residues define a flat and rather hydrophobic collagen binding site located at the front face of the A3 domain. The collagen-binding site of VWF-A3 is distinctly different from that of the homologous integrin alpha(2) I domain, which has a hydrophilic binding site located at the top face of the domain. Based on the surface characteristics of the collagen-binding site of A3, we propose that it interacts with collagen sequences containing positively charged and hydrophobic residues. Docking of a collagen triple helix on the binding site suggests a range of possible engagements and predicts that at most eight consecutive residues in a collagen triple helix interact with A3. PMID- 12582179 TI - The mRNA-binding protein YB-1 (p50) prevents association of the eukaryotic initiation factor eIF4G with mRNA and inhibits protein synthesis at the initiation stage. AB - The cytoplasmic messenger ribonucleoprotein particles of mammalian somatic cells contain the protein YB-1, also called p50, as a major core component. YB-1 is multifunctional and involved in regulation of mRNA transcription and translation. Our previous studies demonstrated that YB-1 stimulates initiation of translation in vitro at a low YB-1/mRNA ratio, whereas an increase of YB-1 bound to mRNA resulted in inhibition of protein synthesis in vitro and in vivo. Here we show that YB-1-mediated translation inhibition in a rabbit reticulocyte cell-free system is followed by a decay of polysomes, which is not a result of mRNA degradation or its functional inactivation. The inhibition does not change the ribosome transit time, and therefore, it affects neither elongation nor termination of polypeptide chains and only occurs at the stage of initiation. YB 1 induces accumulation of mRNA in the form of free messenger ribonucleoprotein particles, i.e. it blocks mRNA association with the small ribosomal subunit. The accumulation is accompanied by eukaryotic initiation factor eIF4G dissociation from mRNA. The C-terminal domain of YB-1 is responsible for inhibition of translation as well as the disruption of mRNA interaction with eIF4G. PMID- 12582181 TI - Hepatitis C virus risk in the interventional radiology environment. PMID- 12582180 TI - An autocatalytic cleavage in the C terminus of the human MUC2 mucin occurs at the low pH of the late secretory pathway. AB - During purification of a recombinant MUC2 C terminus expressed in CHO-K1 cells, the protein was partly cleaved when buffers with a pH of 6.0 were used. When buffers with higher pH values were used, less cleavage was found. Disulfide bonds held the two fragments generated together as these were only observed after reduction. Edman sequencing of the C-terminal 110-kDa fragment revealed that the cleavage had occurred at an Asp-Pro bond, a site described previously to generate the so-called "link peptide" after disulfide bond reduction. In vitro studies on the conditions for cleavage showed that it occurred in a time-dependent manner at a pH below 6.0. Furthermore, the reaction was not enzyme-mediated as it occurred in pure preparations of the MUC2 C terminus and was not inhibited by protease inhibitors. When expressed in the mucin producing cell line LS 174T, the C terminus was cleaved to a higher extent compared with the CHO-K1 cells. Neutralizing the secretory pathway with either NH(4)Cl or bafilomycin A1 inhibited this cleavage. Altogether, our results suggest that the cleavage is an autocatalytic reaction that occurs in the acidic environment of the late secretory pathway. Furthermore, the cleavage produced a new, reactive C terminus that has the potential to attach the mucin to itself or other molecules. Because a pH below 6 can be reached in the late secretory pathway and on mucosal surfaces, the cleavage and possible cross-linking are likely to be of biological importance. PMID- 12582182 TI - Guidelines for the prevention of intravascular catheter-related infections: recommendations relevant to interventional radiology. PMID- 12582183 TI - Selection of stents for treating iliac arterial occlusive disease. AB - Intravascular stents play an increasingly important role in the treatment of iliac artery occlusive disease and their use has expanded the indications for percutaneous endoluminal therapies. The past several years have seen a sharp increase in the number of commercially available covered and uncovered stents. Knowledge of their design and mechanical properties is crucial for selecting the appropriate stent for a particular type of lesion. In this article, the indications for and results of iliac artery stent placement are reviewed and the various characteristics of the currently available stents that may influence operator choice for use in specific lesions are discussed. PMID- 12582185 TI - Potential exposure to hepatitis C virus through accidental blood contact in interventional radiology. AB - PURPOSE: To quantify the prevalence of accidental blood exposure (ABE) among interventional radiologists and contrast that with the prevalence of patients with hepatitis C virus (HCV) undergoing interventional radiology procedures. MATERIALS AND METHODS: A multicenter epidemiologic study was conducted in radiology wards in France. The risk of ABE to radiologists was assessed based on personal interviews that determined the frequency and type of ABE and the use of standard protective barriers. Patients who underwent invasive procedures underwent prospective sampling for HCV serologic analysis. HCV viremia was measured in patients who tested positive for HCV. RESULTS: Of the 77 radiologists who participated in 11 interventional radiology wards, 44% reported at least one incident of mucous membrane blood exposure and 52% reported at least one percutaneous injury since the beginning of their occupational activity. Compliance with standard precautions was poor, especially for the use of protective clothes and safety material. Overall, 91 of 944 treated patients (9.7%) tested positive for HCV during the study period, of whom 90.1% had positive viremia results, demonstrating a high potential for contamination through blood contacts. CONCLUSIONS: The probability of HCV transmission from contact with contaminated blood after percutaneous injury ranged from 0.013 to 0.030; the high frequency of accidental blood exposure and high percentage of patients with HCV could generate a risk of exposure to HCV for radiologists who perform invasive procedures with frequent blood contact. The need to reinforce compliance with standard hygiene precautions is becoming crucial for medical and technical personnel working in these wards. PMID- 12582186 TI - Percutaneous management of ischemic complications in patients with type-B aortic dissection. AB - PURPOSE: To review our experience with the use of percutaneous methods to manage the ischemic complications of type-B aortic dissection. MATERIALS AND METHODS: Retrospective review of our interventional radiology database identified 11 patients with acute type-B aortic dissection who underwent 13 endovascular procedures to attempt revascularization of 23 ischemic vascular territories (four mesenteric, 11 renal, eight lower extremities). Percutaneous interventions included balloon fenestration (four patients), aortic true lumen stent placement (three patients), and branch vessel stent placement (eight patients). RESULTS: Successful initial reperfusion of 21 of 23 vascular territories (91%) was achieved with use of percutaneous methods alone, with no 30-day mortality, in hospital mortality, or paraplegia. One additional patient underwent successful initial treatment with use of a combined interventional/surgical approach to reperfuse an ischemic limb. One patient developed acute thrombosis of a dissected renal artery during stent placement. Increased aortic true lumen collapse later resulted in further ischemia in two patients. During follow-up (mean, 16 mo), no patient has had evidence of false lumen enlargement or late recurrent ischemia. CONCLUSION: Endovascular methods offer a less-morbid nonsurgical treatment alternative for patients with acute complicated type-B aortic dissection. PMID- 12582187 TI - Endovascular treatment of complicated type-B aortic dissection with stent grafts:: midterm results. AB - PURPOSE: To report the midterm results of the endovascular treatment of complicated type-B aortic dissection with use of stent-grafts. MATERIALS AND METHODS: Ten patients with acute (n = 4) or chronic (n = 6) complicated type-B dissection were treated with custom-made stent-grafts. Indications for treatment included uncontrollable hypertension with dissection extension (n = 3), renal ischemia (n = 1), and false lumen aneurysm (n = 6). Stainless-steel Z-stents covered with polyester grafts were placed in the initial six patients. Nitinol stents covered with ultrathin polytetrafluoroethylene were used in the remaining four patients. The patients were followed-up with helical computed tomography for a maximum of 30 months (mean, 20 mo). RESULTS: There was one technical failure related to the access site. Early complications included deep venous thrombosis (n = 1) and embolic stroke (n = 1). Complete thrombosis of the thoracic false lumen was achieved in six patients and partial thrombosis was achieved in three. Aneurysms developed at the ends of the stainless-steel stents in two patients, requiring additional stent-graft placement. Despite successful remodeling of the thoracic aorta, three of four patients with distal reentry into the abdominal aorta experienced progressive abdominal aortic aneurysm (AAA). AAA rupture developed in two patients; one rupture was fatal and the other was treated with emergency surgery. CONCLUSIONS: Endovascular treatment of complicated type-B aortic dissection is technically feasible and effective. Closely monitoring the treated aorta is essential to detect early aneurysm formation at the ends of rigid stents. Despite adequate sealing of the tears in the thoracic aorta, dissection with distal reentry phenomenon into the abdominal aorta may evolve into AAA with late rupture. PMID- 12582188 TI - Early changes in abdominal aortic aneurysm diameter after endovascular repair. AB - PURPOSE: Endovascular repair of abdominal aortic aneurysm (AAA) is expected to alter the natural progression of diameter increase and rupture. The purpose of this study is to determine the rate of diameter change in AAA treated by endovascular repair. MATERIALS AND METHODS: Sixty-three patients underwent endovascular repair of AAA and 12-month median follow-up by computed tomographic (CT) angiography or magnetic resonance (MR) angiography. The maximum cross sectional outer diameters of aneurysms were measured with serial CT angiography and MR angiography. Immediate postrepair CT angiography and MR angiography were used for comparison to follow-up studies. Endoleak was also evaluated. RESULTS: The mean and median follow-up interval was 12 months (range, 7-21 mo). There was a significant decrease in maximum diameter at follow-up (6.0 cm vs 5.1 cm; P <.001). The mean annual decrease of AAA diameter was 8.4 mm. Endoleak occurred immediately after repair in 12 patients (19%). Endoleak was detected in four patients at follow-up examination (6%). Two patients with persistent endoleaks had a mean diameter increase of 2.1 mm per year. Ten patients (16%) with successfully treated endoleak had a mean decrease in diameter of 11 mm per year. There is a significant difference in mean annual diameter change between patients with treated endoleak and those with persistent endoleak (P <.05). There was no difference in mean annual rate of change between patients with no endoleak and those with treated endoleak (8.4 mm/y vs 11 mm/y; P = NS). Seventeen of 21 patients without an appreciable decrease in aneurysm diameter had no endoleak. CONCLUSIONS: Patients with resolved endoleak exhibit a similar shrinkage rate to patients who never had endoleak during imaging follow-up. There remains a group of patients without significant sac shrinkage after endovascular aneurysm repair (EVAR) yet have no endoleak on follow-up imaging (ie, endotension). It is still unclear whether these patients have received protection from AAA rupture from EVAR. PMID- 12582189 TI - Catheter-induced vasospasm in the treatment of acute lower gastrointestinal bleeding. AB - PURPOSE: To demonstrate results in managing lower gastrointestinal (GI) bleeding with the use of superselective catheterization and intentional induction of vasospasm of the bleeding vessel without the use of embolic agents or vasospasm inducing medications. MATERIALS AND METHODS: A retrospective review of 15 episodes of lower GI bleeding treated in the past 6 years by intentional catheter induced vasospasm (CIV) to achieve thrombosis of a bleeding source was conducted. Nine patients had angiographically proven inferior mesenteric artery bleeding and six had angiographically proven superior mesenteric artery bleeding. RESULTS: Bleeding was stopped initially in all patients after effective treatment of the feeding artery. Only one patient experienced a repeat episode of bleeding 2 days later, which required hemicolectomy. Two other patients who underwent adequate embolization underwent surgery at the discretion of the surgeon involved. The remainder were clinically observed and discharged after return of stable vital signs and hematocrit levels. None of the patients treated had clinically evident intestinal ischemia or infarction. There was one significant repeat incidence of bleeding 2 months after CIV that may have represented recurrent bleeding from the original site. CONCLUSION: CIV may be a safe and effective first-line method of embolizing known lower GI bleeding. Whether CIV is used as primary therapy or as the result of spasm incurred during superselective catheterization, the patient may be regarded as successfully treated and followed accordingly, thereby possibly avoiding acute surgical therapy. PMID- 12582190 TI - Renal artery stent placement: complications at a single tertiary care center. AB - PURPOSE: To describe complications after renal artery stent placement in patients with atherosclerotic renal artery stenosis (RAS) at a single tertiary care center and analyze the risk factors for these complications. MATERIALS AND METHODS: Medical records and angiograms of 171 patients who underwent a total of 179 consecutive percutaneous transluminal renal artery stent placement (PTRS) procedures for atherosclerotic RAS were retrospectively reviewed. Data on patient comorbidities, procedure indication, lesion location, serum creatinine level, and procedure-related complications were analyzed. The Wilcoxon rank-sum test was used to assess the association of continuous risk factors with complications, and the chi(2) test was used to assess the association of categoric risk factors with outcomes. RESULTS: The technical success rate for PTRS was 98%. Major complications included renal infarction in five patients (2.8%), permanently increased serum creatinine level in 10 patients (5.6%) that required hemodialysis in five (2.8%), blood transfusion in four patients (2.2%), surgical intervention for procedure-related complications in two patients (1.1%), and deep vein thrombosis in one patient (0.6%). Overall, major complications occurred in 15 procedures (8.4%). Death within 30 days after PTRS occurred in two patients (1.1%), but neither death was definitively caused by the procedure. No risk factors were found to be significantly (P <.05) associated with major complications. Patients with renal artery site or renal function complications tended to be older (P =.01) and have higher creatinine levels (P =.06). CONCLUSIONS: Renal artery stent placement allows treatment of advanced atheromatous RAS in patients with significant medical and surgical comorbidities. Complications are frequent but few have long-term significance. PMID- 12582191 TI - High-resolution MR imaging of human atherosclerotic femoral arteries in vivo: validation with intravascular ultrasound. AB - PURPOSE: To establish a magnetic resonance (MR) imaging protocol for noninvasive in-vivo analysis of atherosclerotic femoral artery segments in humans and to compare the results to those of intravascular ultrasonography (IVUS). MATERIALS AND METHODS: In seven patients with peripheral arterial occlusive disease, 20 femoral arterial segments per person were examined by high-resolution (HR) MR imaging and IVUS. Comparison was possible in 123 of 140 segments. MR imaging was performed at 1.5 T with use of a three-dimensional (3D) time-of-flight sequence with an in-plane resolution of 0.78 x 0.49 mm(2). 3D contrast-enhanced MR angiography was used for exact positioning of the HR MR imaging slices. IVUS (3.5 F, 40 MHz) was performed with use of a motorized pullback system. Parameters analyzed included cross-sectional lumen area (LA), vessel area (VA), and extent of vessel wall calcification. RESULTS: Agreement between IVUS and HR MR imaging was analyzed with use of the Bland-Altman method. The paired LA measurements were in close agreement: the Bland-Altman mean bias in LA was -0.4 mm with a precision of +/-5.1 mm (P =.062). As a result of dorsal echo extinction in IVUS, VA measurements were feasible in only 74 of 140 segments. VA measurements were moderately correlated (r = 0.74; P <.0001), and a 25% overestimation by HR MR imaging compared to IVUS was observed. Intra- and interobserver comparisons for LA and VA measured with HR MR imaging did not show significant differences. Vessel wall calcifications were classified with a sensitivity of 91%, a specificity of 93%, and an accuracy of 93%. CONCLUSIONS: The MR imaging protocol introduced in the present study permits precise assessment of LA and extent of calcification in peripheral arterial occlusive disease in vivo. HR MR imaging shows high concordance with IVUS and may have the potential for noninvasive therapy monitoring. PMID- 12582192 TI - Endothelial dysfunction after arterial thrombosis is ameliorated by L-arginine in combination with thrombolysis. AB - PURPOSE: To assess endothelial function after arterial thrombosis creation and after administration of a novel thrombolytic regimen in a new porcine model. MATERIALS AND METHODS: Untreated arteries that had undergone thrombosis for 90 minutes were compared to arteries treated with tissue plasminogen activator (tPA, 4 mg) and a combination of tPA and L-arginine (L-arg; 600 mmol/L). External iliac artery luminal diameter was measured with use of duplex ultrasonography. Endothelial-dependent relaxation (EDR) and endothelial-independent relaxation (EIR) were measured with use of acetylcholine chloride (ACh) and sodium nitroprusside (NTP), respectively. Endothelial integrity was confirmed by scanning electron microscopy (SEM). Nitric oxide (NO) levels were determined with use of a chemiluminescent assay of its nitrite/nitrate metabolites (NO(x)). RESULTS: After thrombosis, EDR was decreased (69% +/- 9.5; ACh = 15 micro g/min; n = 6). EDR remained unchanged after thrombolysis with tPA despite complete dissolution of thrombus (67% +/- 5.7; ACh = 15 micro g/min; n = 5). Thrombolysis with use of tPA coupled with L-arg resulted in an increase in EDR (95% +/- 4.9; ACh = 15 micro g/min; n = 5; P =.007). EIR was preserved in all groups, with uniform response to NTP. SEM analysis revealed intact endothelium in all groups. Local NO(x) levels were diminished after 90 minutes of thrombosis (49.3 micro mol/L vs 40.8 micro mol/L; P =.0002), but increased to 55.7 micro mol/L after thrombolysis with tPA and L-arg (P = NS). CONCLUSIONS: Thrombus induces arterial dysfunction acutely without altering endothelial integrity. This dysfunction is ameliorated through regional administration of L-arg in combination with standard thrombolytic therapy, which increases local NO levels. This model allows the in vivo study of thrombosis and alternative thrombolytic regimens. Regional enhancement of NO levels may prove to be an attractive adjunct in thrombolytic therapy. PMID- 12582193 TI - Radiofrequency ablation with and without balloon occlusion of the renal artery: an experimental study in porcine kidneys. AB - PURPOSE: To compare thermal lesion volumes and associated renal damage (other than thermal lesions) when radiofrequency (RF) ablation is performed with and without balloon occlusion of the renal artery in normal porcine kidneys. MATERIALS AND METHODS: RF ablation was performed in six pigs. An electrode with eight circumferential prongs arranged in a circle 20 mm in diameter was inserted into the lower pole of the kidney. In each animal, renal ablation was performed with balloon occlusion of the renal artery in one kidney and without balloon occlusion in the contralateral kidney. The volume of the thermal lesion and associated renal damage other than the thermal lesion were compared by angiography and histology between the groups of kidneys ablated with and without arterial occlusion. The volume of thermal lesion was measured by tracing the thermal lesions of sliced kidneys and multiplying each area by the slice thickness (5 mm). RESULTS: Renal ablation with arterial occlusion consistently created nearly spherical thermal lesions with a mean volume of 5,399 mm(3) +/- 902. RF ablation without arterial occlusion, conversely, created irregular, mushroom-shaped thermal lesions limited to the region around the prongs and shaft of the electrode, with a mean volume of 2,759 mm(3) +/- 1,069 (P <.001). Renal arteriography showed occlusion of interlobar arteries penetrating the thermal lesions and perfusion defects peripheral to the thermal lesions immediately after RF ablation. Perfusion defects were larger in kidneys ablated with arterial occlusion than in those ablated without arterial occlusion (1,499 cm(2) +/- 354 vs 499 cm(2) +/- 352; P <.001). CONCLUSIONS: Renal RF ablation with arterial occlusion consistently provides larger thermal lesions than ablation without arterial occlusion does, but it induces renal infarction peripheral to the thermal lesion. PMID- 12582194 TI - Use of internal coils for independent and direct MR imaging-guided endovascular device tracking. AB - PURPOSE: To test the hypotheses that a single internal guide wire coil (i) permits independent and direct depiction of guide wires and catheters and (ii) improves catheter-tracking accuracy and depiction compared to external receiver coils. MATERIALS AND METHODS: Standard 5-6-F angiographic catheters were filled with dilute 4% gadolinium chelate. A single 0.030-inch-diameter internal guide wire coil was placed inside the catheter. True fast imaging with steady-state precession was used to directly visualize the guide wire. Inversion recovery prepared fast low-angle shot technique was used to track catheters over a thick slice. In phantom experiments, we compared catheter signal-to-noise ratios (SNRs) with the internal coil and a phased-array surface coil with use of the Wilcoxon signed-rank test. Tip-tracking accuracy was assessed with use of linear regression. In pigs (n = 7), catheters and guide wires were independently tracked in real time. RESULTS: In phantoms, catheter SNR with the internal coil (12.0) was significantly greater than that with the surface coil (4.0; P =.001). Tip tracking accuracy was also improved with use of the internal coil (R(2) = 0.94 vs 0.50). In swine vasculature, catheters and guide wires could be directly and independently tracked at 1.7-2.0 frames per second. Catheters were clearly visualized with use of the internal coil, with a typical catheter background contrast-to-noise ratio of 6.6. Catheters were not visible with use of the external coil because of the small catheter size compared to the slice thickness. CONCLUSION: Internal guide wire coils permit independent and direct depiction of guide wires and catheters in vivo for MR imaging-guided endovascular interventions. They also improve catheter tracking accuracy and depiction compared to external coils. PMID- 12582195 TI - Pericardial tamponade complicating central venous interventions. AB - Cardiac tamponade is defined as an accumulation of fluid within the pericardial sac that is large enough to impair the diastolic filling of the ventricles. There are numerous causes, including cardiac disease, malignancy, and trauma (penetrating, blunt, and iatrogenic). With increasing complexity of percutaneous endovascular interventions, interventional radiologists must learn to recognize the preexisting and iatrogenic presence of cardiac tamponade during procedures and be familiar with its treatment. Two cases of pericardial tamponade complicating central venous interventions are described. In addition, the clinical condition is discussed and its diagnosis and treatment are reviewed. PMID- 12582196 TI - Percutaneous cholecystocholedochostomy for cholecystitis and cystic duct obstruction in gallbladder carcinoma. AB - Percutaneous cholecystocholedochostomy was performed in a patient with acute cholecystitis caused by cystic duct obstruction by gallbladder carcinoma, but removal of the percutaneous cholecystostomy catheter was unsuccessful because of continuing discharge. After creation of a cholecystocholedochostomy through the cholecystostomy tract with use of a transjugular liver access set and a 21-gauge needle, self-expandable metallic stents were placed in the narrowed common bile duct and the newly created tract between the gallbladder and the common hepatic duct. The external cholecystostomy catheter was successfully removed after the procedure. Jaundice occurred 70 days later as a result of tumor invasion above the segment with the stent, and an additional stent was placed. The patient died of diffuse metastasis 143 days after creation of the cholecystocholedochostomy without recurrence of cholecystitis. PMID- 12582197 TI - Placement of 10-millimeter Trufill platinum microcoils in the gastroduodenal artery for intraarterial implantation of a microcatheter-port system to treat liver tumors. PMID- 12582198 TI - Focus issue: the signaling skinny on obesity. PMID- 12582199 TI - A long search for Glut4 activation. AB - Insulin stimulates glucose transport in its target cells by translocation of the glucose transporter isoform 4 (Glut4) from an intracellular storage pool to the plasma membrane. A large body of evidence indicates that activity of Glut4 at the plasma membrane may vary. Recent findings suggest that p38 MAPK may be involved in regulation of the intrinsic activity of the transporter. PMID- 12582200 TI - A new role for SOCS in insulin action. Suppressor of cytokine signaling. AB - Suppressor of cytokine signaling (SOCS) family proteins were initially identified as inhibiting cytokine signaling pathways through a negative feedback loop involving the inhibition of Janus kinase activity. More recent data suggest that SOCS proteins may also modulate signaling mediated through receptor tyrosine kinases. Krebs and Hilton discuss research implicating SOCS-1 and SOCS-3 as inhibitors of insulin receptor-mediated and insulin-like growth factor-1 receptor mediated signaling pathways, as well as the increasing evidence that SOCS proteins may act in part by participating in a ubiquitin ligase complex to promote the degradation of target proteins. PMID- 12582201 TI - Leptin--a critical body weight signal and a "master" hormone? AB - Leptin, initially identified as a hormone produced by white fat that acted as a satiety signal, has since been found to be synthesized in various tissues and to subserve diverse functions, including regulation of blood glucose concentrations and blood vessel growth, and signaling to the reproductive and immune systems. Recent research suggests that leptin, acting through the sympathetic nervous system, may also regulate bone density. Trayhurn discusses this research in the context of the established bidirectional interaction between leptin and the sympathetic nervous system, and the need to elucidate a unifying theme with which to make sense of leptin's myriad, seemingly quite disparate, functions. PMID- 12582202 TI - Single cell imaging of PI3K activity and glucose transporter insertion into the plasma membrane by dual color evanescent wave microscopy. AB - Many signaling events involve the translocation of signaling molecules to or from the plasma membrane; however, suitable techniques to quantify the temporal relationships between such signaling events are lacking. Here, we describe an evanescent wave microscopy technique that allows parallel measurement of the recruitment and dissociation of cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) labeled proteins to and from the plasma membrane in individual living cells. The selective excitation of fluorescence in a zone less than 100 nm above a cover glass enables selective imaging within the plasma membrane of adherent cells, with markedly improved resolution, lower background, and minimal phototoxicity compared to confocal microscopy and other microscopy based assays. In the microscope design we have developed, the beams from helium cadmium (442 nm) and argon (514 nm) lasers are merged and focused through a dove prism at an angle that yields total internal reflection. In this configuration, evanescent wave-excited fluorescence at the glass-water interface can be detected with either high or low magnification, to allow for high-resolution imaging or the study of many cells in parallel. We applied this technique to make parallel measurements of the time-course of insulin-triggered activation of phosphatidylinositol 3-kinase (PI3K) and GLUT4 glucose transporter insertion into the plasma membrane of individual differentiated 3T3L1 adipocytes using a phosphatidylinositol-3,4,5-trisphosphate [PI(3,4,5)P(3)]-binding pleckstrin homology domain fused to CFP, and GLUT4 conjugated to YFP. The technique should have wide applicability to various cell types and diverse signaling processes. PMID- 12582203 TI - Alkaline phosphatase fusion proteins as affinity probes for protein localization studies. AB - The receptor affinity probe or receptor alkaline phosphatase (RAP) staining method uses soluble protein ectodomains fused to secreted placental alkaline phosphatase to locate ectodomain binding sites within cells or tissues. We have used this approach to identify expressing cells in tissue culture, in tissue sections, or in whole-mount embryos. The RAP method is especially useful in situations where a reliable monoclonal antibody is not available or if an orphan receptor is the focus of the study. The technique permits localization of both receptors and ligands and is readily quantifiable for cell-surface binding assays. Soluble ectodomain placental alkaline phosphatase fusion proteins are therefore highly sensitive reagents that permit the direct localization of available binding sites through simple chromogenic assays without purification, radioactive labeling, or secondary reagents. PMID- 12582204 TI - Determination of cellular H2O2 production. AB - The detection of radical oxygen species (ROS) is central to the understanding of their role in signal transduction. ROS detection is based on the radical dependent reduction of a compound with a measurable change in a chemical property. However, in vitro several of the compounds are reduced by several oxidants, which results in a lack of specificity when used to detect intracellular ROS. In contrast, by using methods that detect extracellular ROS, specificity can be corroborated by adding the appropriate competitor; for example, superoxide dismutase competes for superoxide, and catalase competes for H(2)O(2). In addition, because of the efficient activity of superoxide dismutases on superoxide and H(2)O(2) membrane permeability, determination of extracellular H(2)O(2) can detect all potential cellular sources of these ROS. In this protocol, extracellular H(2)O(2 )is measured as the limiting factor of peroxidase mediated oxidation of homovanillic acid into a fluorescent dimer. The specificity of this reaction for H(2)O(2) is demonstrated by the addition of catalase as an H(2)O(2) scavenger. Because the assay detects small changes in fluorescence, it is highly sensitive. The high sensitivity and the specificity of this assay make it well suited to measure ROS in nonphagocytic cells where the ROS levels are in the low micromolar range. To further increase sensitivity, H(2)O(2) measurements are performed over time to ensure the detection of maximum response and to minimize the variability in response arising from cellular heterogeneity, an attribute of primary cultures. PMID- 12582205 TI - Nicotine activates immature "silent" connections in the developing hippocampus. AB - In the hippocampus at birth, most glutamatergic synapses are immature and functionally "silent" either because the neurotransmitter is released in insufficient amount to activate low-affinity alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionate receptors or because the appropriate receptor system is missing or nonfunctional. Here we show that, in the newborn rat, a brief application of nicotine at immature Schaffer collateral-CA1 connections strongly enhances neurotransmitter release and converts presynaptically silent synapses into conductive ones. This effect is persistent and can be mimicked by endogenous acetylcholine released from cholinergic fibers. Thus, during a critical period of postnatal development, activation of nicotinic acetylcholine receptors contributes to the maturation of functional synaptic contacts and the wiring of adult hippocampal circuitry. PMID- 12582206 TI - Binding of small molecules to an adaptive protein-protein interface. AB - Understanding binding properties at protein-protein interfaces has been limited to structural and mutational analyses of natural binding partners or small peptides identified by phage display. Here, we present a high-resolution analysis of a nonpeptidyl small molecule, previously discovered by medicinal chemistry [Tilley, J. W., et al. (1997) J. Am. Chem. Soc. 119, 7589-7590], which binds to the cytokine IL-2. The small molecule binds to the same site that binds the IL-2 alpha receptor and buries into a groove not seen in the free structure of IL-2. Comparison of the bound and several free structures shows this site to be composed of two subsites: one is rigid, and the other is highly adaptive. Thermodynamic data suggest the energy barriers between these conformations are low. The subsites were dissected by using a site-directed screening method called tethering, in which small fragments were captured by disulfide interchange with cysteines introduced into IL-2 around these subsites. X-ray structures with the tethered fragments show that the subsite-binding interactions are similar to those observed with the original small molecule. Moreover, the adaptive subsite tethered many more compounds than did the rigid one. Thus, the adaptive nature of a protein-protein interface provides sites for small molecules to bind and underscores the challenge of applying structure-based design strategies that cannot accurately predict a dynamic protein surface. PMID- 12582207 TI - Desensitization, internalization, and signaling functions of beta-arrestins demonstrated by RNA interference. AB - Beta-arrestins bind to activated G protein-coupled receptor kinase-phosphorylated receptors, which leads to their desensitization with respect to G proteins, internalization via clathrin-coated pits, and signaling via a growing list of "scaffolded" pathways. To facilitate the discovery of novel adaptor and signaling roles of beta-arrestins, we have developed and validated a generally applicable interfering RNA approach for selectively suppressing beta-arrestins 1 or 2 expression by up to 95%. Beta-arrestin depletion in HEK293 cells leads to enhanced cAMP generation in response to beta(2)-adrenergic receptor stimulation, markedly reduced beta(2)-adrenergic receptor and angiotensin II receptor internalization and impaired activation of the MAP kinases ERK 1 and 2 by angiotensin II. This approach should allow discovery of novel signaling and regulatory roles for the beta-arrestins in many seven-membrane-spanning receptor systems. PMID- 12582208 TI - Patterns of global gene expression in rat skeletal muscle during unloading and low-intensity ambulatory activity. AB - Physical inactivity and unloading lead to diverse skeletal muscle alterations. Our goal was to identify the genes in skeletal muscle whose expression is most sensitive to periods of unloading/reduced physical activity and that may be involved in triggering initial responses before phenotypic changes are evident. The ability of short periods of physical activity/loading as an effective countermeasure against changes in gene expression mediated by inactivity was also tested. Affymetrix microarrays were used to compare mRNA levels in the soleus muscle under three experimental treatments (n = 20-29 rats each): 12-h hindlimb unloading (HU), 12-h HU followed by 4 h of intermittent low-intensity ambulatory and postural activity (4-h reloading), and control (with ambulatory and postural activity). Using a combination of criteria, we identified a small set of genes (approximately 1% of 8,738 genes on the array or 4% of significant expressed genes) with the most reproducible and largest responses to altered activity. Analysis revealed a coordinated regulation of transcription for a large number of key signaling proteins and transcription factors involved in protein synthesis/degradation and energy metabolism. Most (21 of 25) of the gene expression changes that were downregulated during HU returned at least to control levels during the reloading. In surprising contrast, 27 of 38 of the genes upregulated during HU remained significantly above control, but most showed trends toward reversal. This introduces a new concept that, in general, genes that are upregulated during unloading/inactivity will be more resistant to periodic reloading than those genes that are downregulated. This study reveals genes that are the most sensitive to loading/activity in rat skeletal muscle and indicates new targets that may initiate muscle alterations during inactivity. PMID- 12582209 TI - PDK4: A factor in fatness? PMID- 12582210 TI - Effects of growth hormone administration in human obesity. AB - OBJECTIVE: To summarize the reports in the literature regarding the effect of growth hormone (GH) treatment of obesity. RESEARCH METHODS AND PROCEDURES: Clinical trials of GH treatment of obese adults were reviewed and summarized. Specifically, information regarding the effects of GH on body fat and body fat distribution, glucose tolerance/insulin resistance, and adverse consequences of treatment were recorded. RESULTS: GH administered together with hypocaloric diets did not enhance fat loss or preserve lean tissue mass. No studies provided strong evidence for an independent beneficial effect of GH on visceral adiposity. In all but one study, glucose tolerance during GH treatment suffered relative to placebo. CONCLUSION: The bulk of studies indicate little or no beneficial effects of GH treatment of obesity despite the low serum GH concentrations associated with obesity. PMID- 12582211 TI - Reduced PDK4 expression associates with increased insulin sensitivity in postobese patients. AB - OBJECTIVE: The aim of this study was to verify whether changes in PDK4 mRNA expression in skeletal muscle in formerly obese subjects who underwent malabsorptive bariatric surgery [bilio-pancreatic diversion (BPD)] might be related to insulin sensitivity improvement, and if these possible modifications might correlate with a reduction of the intramyocytic lipid level. RESEARCH METHODS AND PROCEDURES: Six obese women (body mass index 46.6 +/- 8.2 kg/m(2)) were enrolled in the study. Body composition, euglycemic-hyperinsulinemic clamp and muscle biopsies for skeletal muscle lipid analysis, and semiquantitative reverse transcriptase-polymerase chain reaction were performed before and 3 years after BPD. RESULTS: The average weight loss observed after surgery was approximately 42%. Increased glucose uptake was accompanied by a significant decrease of PDK4 mRNA (R(2) = 0.71, p < 0.001). The amounts of intramyocytic triglycerides correlate directly with PDK4 mRNA (R(2) = 0.87, p = 0.005) and inversely with glucose uptake values (R(2) = 0.75, p < 0.001). DISCUSSION: Our results support the concept that a reduced tissue availability of fatty acids consequent to a massive lipid malabsorption influences glucose metabolism acting through the regulation of PDH complex. In fact, as shown in animals, a higher level of FFA availability is likely to induce overexpression of PDK4 also in humans. PMID- 12582212 TI - Effect of fat sources on satiety. AB - OBJECTIVE: There are inconsistent reports on the satiety value of different fatty acids. This study compared the appetitive effects of two fat sources rich in monounsaturated fatty acids (peanut oil and canola oil) with a source rich in saturated fatty acids (butter). RESEARCH METHODS AND PROCEDURES: After an overnight fast, lean participants completed a questionnaire eliciting information about hunger, fullness, desire to eat, and prospective consumption. They then consumed one of the preloads (muffins containing 40 g of each fat source or no fat) and 150 mL of water within 15 minutes. Questionnaires were completed again 30, 60, and 120 minutes after preload ingestion. Participants kept dietary records during the subsequent 24 hours. RESULTS: Canola and peanut oil muffins resulted in higher fullness, and butter, canola, and peanut oil muffins resulted in lower hunger ratings 30, 60, and 120 minutes after preload ingestion compared with the fat-free preload. No differences were observed among the fat-containing loads. Although energy intake 24 hours after consumption of the preloads was also comparable on days the three fat-containing loads were consumed, energy consumption after each study session was higher when the fat-free muffins were provided. However, total energy intake, including the calories provided by the preloads, was similar across treatments. DISCUSSION: These data do not support a differential satiety effect of fat sources rich in monounsaturated fatty acids relative to one rich in saturated fatty acids. PMID- 12582213 TI - DNA microarray analysis of genes differentially expressed in diet-induced (cafeteria) obese rats. AB - OBJECTIVE: To better understand the molecular basis of dietary obesity, we examined adipose tissue genes differentially expressed in an obesity model using DNA microarray analysis. RESEARCH METHODS AND PROCEDURES: We assessed the expression level of over 12,500 transcripts in epididymal fat pads from (cafeteria) obese and control rats with the aid of the array technology. RESULTS: Cafeteria (obese) rats weighed 50% more and had 2.5-fold higher levels of epididymal fat and elevated levels of circulating leptin. Adipose genes differentially expressed in obese and control rats were categorized into five groups: macronutrient metabolism, transcription factors, hormone receptor and signal transduction, redox and stress proteins, and cellular cytoskeleton. Interestingly, the expression levels of a number of genes involved in lipid metabolism such as glycerol-3-phosphate dehydrogenase, stearoyl coenzyme A desaturase, together with the transcription factors implicated in adipocyte differentiation (CAAT/enhancer binding protein-alpha and peroxisome proliferator activated receptor-gamma), were significantly increased in obese animals compared with control. The most up-regulated transcripts were the ob (49.2-fold change) and the fatty acid-binding protein genes (15.7- fold change). In contrast, genes related to redox and stress protein were generally down-regulated in obese animals compared with the control. DISCUSSION: Our study showed that in diet induced obesity, the expression levels of some important genes implicated in lipid metabolism were up-regulated, whereas those related to redox and stress protein were down-regulated in obese animals compared with control. This pattern of gene expression may occur in human obesity cases after high-fat intake. PMID- 12582214 TI - Emotional eating, alexithymia, and binge-eating disorder in obese women. AB - OBJECTIVE: To investigate the relationships between alexithymia and emotional eating in obese women with or without Binge Eating Disorder (BED). RESEARCH METHODS AND PROCEDURES: One hundred sixty-nine obese women completed self-report questionnaires, including the Beck Depression Inventory, the State Trait Anxiety Inventory, the Stress Perceived Scale, the Dutch Eating Behaviour Questionnaire, and the Toronto Alexithymia Scale. The presence of BED, screened using the Questionnaire of Eating and Weight Patterns, was confirmed by interview. RESULTS: Forty obese women were identified as having BED. BED subjects and non-BED subjects were comparable in age, body mass index, educational level, and socioeconomic class. According to the Dutch Eating Behaviour Questionnaire, BED subjects exhibited higher depression, anxiety, perceived stress, alexithymia scores, and emotional and external eating scores than non-BED subjects. Emotional eating and perceived stress emerged as significant predictors of BED. The relationships between alexithymia and emotional eating in obese subjects differed between the two groups according to the presence of BED. Alexithymia was the predictor of emotional eating in BED subjects, whereas perceived stress and depression were the predictors in non-BED subjects. DISCUSSION: This study pointed out different relationships among mood, alexithymia, and emotional eating in obese subjects with or without BED. Alexithymia was linked to emotional eating in BED. These data suggest the involvement of alexithymia in eating disorders among obese women. PMID- 12582215 TI - Birth weight, adult body composition, and subcutaneous fat distribution. AB - OBJECTIVES: To investigate if birth weight is related to both body mass index (BMI) and distribution of subcutaneous fat at adult age. RESEARCH METHODS AND PROCEDURES: A 9-year longitudinal study was performed in 229 subjects (192 women) with ages ranging from 27 to 36 years. Birth weight was retrieved by a questionnaire, and adult weight, height, skinfold thicknesses, and waist-to-hip ratio (WHR) were repeatedly measured at mean ages 27, 29, 31, and 36 years. BMI, sum of four skinfolds (S4S), the ratio between two truncal skinfolds and S4S (SS/S4S), and the ratio between WHR and the cross-sectional area of the left thigh were calculated with the available data. RESULTS: The adjusted model showed that in women, birth weight was significantly negatively related to adult S4S [beta = -5.211; (-9.768 to -0.654)], waist circumference [beta = -1.449; (-2.829 to -0.069)], and SS/S4S ratio [beta = -3.579; (-5.296 to -1.862)]. In men, a significant negative association was observed between birth weight and adult WHR [beta = -1.096; (-2.092 to -0.100)] only. Other relationships showed, although not significantly, the same negative trend, namely that lower birth weight is related to higher adult body fat mass (S4S) and a more truncal subcutaneous fat distribution (SS/S4S). No associations were found between birth weight and either adult BMI or the cross-sectional area of the thigh. DISCUSSION: Lower birth weight is, in both adult men and women, related to a higher adult subcutaneous fat mass and a more truncal distribution of subcutaneous fat, indicating a higher risk for obesity. PMID- 12582216 TI - Obesity and quality of life: mediating effects of pain and comorbidities. AB - OBJECTIVE: To estimate the association between body mass index (BMI) and health related quality of life (HRQL) and examine whether joint pain and obesity-related comorbidities mediate the BMI-HRQL association. RESEARCH METHODS AND PROCEDURES: Population-based survey data from the 1999 Behavioral Risk Factor Surveillance Survey. Adults (N = 155,989) were classified according to BMI as underweight (<18.5 kg/m(2)), desirable weight (18.5 to 24.9 kg/m(2)), overweight (25 to 29.9 kg/m(2)), obese class I (30 to 34.9 kg/m(2)), obese class II (35 to 39.9 kg/m(2)), and obese class III (>/=40 kg/m(2)). Data including general health status, unhealthy days in the past 30 caused by physical problems and mental problems, and total unhealthy days in the past 30 were collected. RESULTS: After adjusting for age, sex, race, smoking, education, and income, we observed J shaped associations between BMI and HRQL. Compared with desirable weight adults, underweight, overweight, and obesity classes I, II, and III adults [odds ratio (OR) = 1.57, 1.19, 1.95, 2.72, and 4.36, respectively] were significantly (p < 0.001) more likely to report fair/poor general health status. For unhealthy days caused by physical problems, the corresponding ORs were 1.51, 1.15, 1.66, 2.27, and 3.61 (p < 0.001). For unhealthy days caused by mental problems, the ORs were 1.35, 1.14 1.43, 1.57, and 2.25 (p < 0.001). For total unhealthy days, the corresponding ORs were 1.27, 1.09, 1.37, 1.73, and 2.46 (p < 0.01). Adding joint pain and obesity-related comorbidities into models attenuated BMI-HRQL associations. DISCUSSION: Associations between BMI and HRQL indices were J shaped. Joint pain and comorbidities may mediate BMI-HRQL associations. PMID- 12582217 TI - Volume estimates by imaging methods: model comparisons with visible woman as the reference. AB - OBJECTIVE: To compare the accuracy of four volume estimation models to actual tissue and organ volumes measured in the visible woman. METHODS: Actual volumes were calculated from 1-mm-thick visible woman images that were segmented for five major components including subcutaneous and visceral adipose tissue across the 1730 available slices. Four available models resolved to two equations: truncated cone/truncated pyramid vs. two-column/parallel trapezium. Between-slice interval and initial slice were systematically varied when deriving component volumes using the two equations in four regions. RESULTS: For each compartment and each between-slice interval, the means of the two-column model were always the same as the corresponding reference volumes, whereas those of the truncated cone model were smaller than the reference volumes. Similarly, the coefficient variation for the two-column model was always smaller than for the truncated cone model. DISCUSSION: The equation based on the parallel trapezium and the two-column models is more accurate in estimating tissue volumes than the corresponding equation for truncated pyramid and truncated cone models. This finding has important implications for the volume calculations of imaging-based body compartments such as adipose tissue. PMID- 12582218 TI - Relationship of neck circumference to cardiovascular risk factors. AB - OBJECTIVE: To determine a relationship between neck circumference (NC) and risk factors for coronary heart disease by evaluating the components of the metabolic syndrome. RESEARCH METHODS AND PROCEDURES: The study group included 561 subjects (231 men and 330 women) who had no known major medical conditions and were not receiving any medication therapy. The subjects were those who attended a family health clinic for any reason between 1998 and December 2001. Main indicators studied included NC, waist circumference, waist-to-hip ratio, body mass index, blood pressure, and lipoprotein, glucose, and uric acid levels. RESULTS: Pearson's correlation coefficients indicated a significant association between NC and body mass index (men, r = 0.71; women, r = 0.81; each, p < 0.0001), waist circumference (men, r = 0.75; women, r = 0.79; each, p < 0.0001), waist-to-hip ratio (men, r = 0.56; women, r = 0.63; each, p < 0.0001), total cholesterol (men, r = 0.50; women, r = 0.66; each, p < 0.0001), low-density lipoprotein-cholesterol (men, r = 0.42; women, r = 0.60; each, p < 0.0001), triglycerides (men, r = 0.48; women, r = 0.49; each, p < 0.0001), glucose (men, r = 0.21, p < 0.001; women, r = 0.44; p < 0.0001), uric acid (men, r = 0.50, p < 0.0001; women, r = 0.60, p < 0.001), and systolic (men, r = 0.53; women, r = 0.69; each, p < 0.0001), and diastolic (men, r = 0.55; women, r = 0.65; each, p < 0.0001) blood pressure. DISCUSSION: Higher NC is correlated positively with the factors of the metabolic syndrome; therefore, it is likely to increase the risk of coronary heart disease. PMID- 12582219 TI - Leptin response to glucocorticoid occurs at physiological doses and is abolished by fasting. AB - OBJECTIVE: To examine the effects of graded doses of hydrocortisone (HC) on leptin secretion, and determine the effect of fasting. RESEARCH METHODS AND PROCEDURES: This was a randomized, placebo-controlled, crossover study, with a 1 week "washout" period between interventions. Eight healthy subjects [age = 36 +/- 2.3 years (+/-SE), body mass index = 31.5 +/- 1.6 kg/m(2)] completed the dose response study in which an intravenous infusion of saline (placebo) or HC (30 or 100 mg) was administered for 24 hours. Four healthy subjects (age = 35.2 +/- 3.0 years, body mass index = 27.1 +/- 2.1 kg/m(2)) completed the fasting study, which entailed continuous infusion of saline, HC (300 mg/24 hours) in the fed state, or HC (300 mg/24 hours) with total caloric deprivation for 24 hours. Blood sampling was performed every 1 to 2 hours for measurement of leptin, cortisol, insulin, and glucose levels. RESULTS: Peak hyperleptinemia occurred after 16 hours of HC infusion; peak/baseline leptin levels were 129% (placebo), 140% (30 mg of HC for 24 hours, p = 0.05), and 185% (100 mg of HC for 24 hours, p < 0.01). During infusion of HC (300 mg/24 hours or placebo), the peak/baseline plasma leptin levels were 16.1 +/- 5.8/12.8 +/- 5.9 ng/mL (placebo with food, 126%), 14.6 +/- 6.0/12.5 +/- 6.5 ng/mL (HC fasting, 117%), and 32.5 +/- 12.5/12.0 +/- 8.4 ng/mL (HC with food, 271%, p < 0.001). DISCUSSION: Leptin secretory responses occur at physiological doses of HC, are obliterated by fasting, and thus may be of metabolic significance. PMID- 12582220 TI - Overweight children's barriers to and support for physical activity. AB - OBJECTIVE: As the epidemic of overweight increases among youth, research needs to examine factors that may influence children's participation in weight-related health behaviors. This study examined overweight children's perceived barriers to and support for physical activity compared with nonoverweight children. RESEARCH METHODS AND PROCEDURES: Barriers to and support for physical activity were examined among 84 overweight children attending a summer fitness camp or a university-based weight loss clinic. Barriers and support levels were then compared with those of 80 nonoverweight children of a similar age range. RESULTS: Body-related barriers were the most predominant barrier type among overweight youth, especially among overweight girls. Overweight children, particularly girls, reported significantly higher body-related, resource, and social barriers to physical activity compared with nonoverweight children and lower levels of adult support for physical activity. DISCUSSION: Overweight children may be particularly vulnerable to body-related barriers to physical activity, and reducing such barriers may serve as physical activity intervention points most relevant for overweight youth. Future interventions may also benefit from enhancing support for physical activity from adults and peers. PMID- 12582221 TI - Effects of a longitudinal training program on responses to exercise in overweight men. AB - OBJECTIVE: The aim of this study was to determine how training modifies metabolic responses and lipid oxidation in overweight young male subjects. RESEARCH METHODS AND PROCEDURES: Eleven overweight subjects were selected for a 4-month endurance training program. Before and after the training period, they cycled for 60 minutes at 50% of their VO(2)max after an overnight fast or 3 hours after eating a standardized meal. Various metabolic and endocrine parameters, and respiratory exchange ratio values were evaluated. RESULTS: Exercise-induced plasma norepinephrine concentration increases were similar before and after training in fasted or fed conditions. After food intake, exercise promoted a decrease in plasma glucose and a higher increase in epinephrine than in fasting conditions. The increase in epinephrine after the meal was more marked after training (264 +/ 32 vs. 195 +/- 35 pg/mL). Training lowered the resting plasma nonesterified fatty acids. During exercise, changes in glycerol were similar to those found before training. Lipid oxidation during exercise was higher in fasting than in fed conditions (15.5 +/- 1.4 vs. 22.3 +/- 1.7 g/h). Training did not significantly increase fat oxidation when exercise was performed in fed conditions, but it did in fasting conditions (18.6 +/- 1.4 vs. 27.2 +/- 1.8 g/h). DISCUSSION: Endurance training decreased plasma nonesterified fatty acids, cholesterol, and insulin concentrations. Training increased lipid oxidation during exercise, in fasting conditions, and not when exercise was performed after the meal. During exercise in overweight subjects, the fasting condition seems more suited to oxidizing fat and maintaining glucose homeostasis than a 3-hour wait after a standard meal. PMID- 12582222 TI - Insulin- and exercise-stimulated skeletal muscle blood flow and glucose uptake in obese men. AB - OBJECTIVE: Insulin resistance in obese subjects results in the impaired use of glucose by insulin-sensitive tissues, e.g., skeletal muscle. In the present study, we determined whether insulin resistance in obesity is associated with an impaired ability of exercise to stimulate muscle blood flow, oxygen delivery, or glucose uptake. RESEARCH METHODS AND PROCEDURES: Nine obese (body mass index = 36 +/- 2 kg/m(2)) and 11 age-matched nonobese men (body mass index = 22 +/- 1 kg/m(2)) performed one-legged isometric exercise during hyperinsulinemia. Rates of femoral muscle blood flow, oxygen consumption, and glucose uptake were measured simultaneously in both legs using [(15)O]H(2)O, [(15)O]O(2), [(18)F]fluoro-deoxy-glucose, and positron emission tomography. RESULTS: The obese subjects exhibited resistance to insulin stimulation of glucose uptake in resting muscle, regardless of whether glucose uptake was expressed per kilogram of femoral muscle mass (p = 0.001) or per the total mass of quadriceps femoris muscle. At similar workloads, oxygen consumption, blood flow, and glucose uptake were lower in the obese than the nonobese subjects when expressed per kilogram of muscle, but similar when expressed per quadriceps femoris muscle mass. DISCUSSION: We conclude that obesity is characterized by insulin resistance of glucose uptake in resting skeletal muscle regardless of how glucose uptake is expressed. When compared with nonobese individuals at similar absolute workloads and under identical hyperinsulinemic conditions, the ability of exercise to increase muscle oxygen uptake, blood flow, and glucose uptake per muscle mass is blunted in obese insulin-resistant subjects. However, these defects are compensated for by an increase in muscle mass. PMID- 12582223 TI - A genome-wide scan for body mass index among Nigerian families. AB - OBJECTIVE: Interest in mapping genetic variants that are associated with obesity remains high because of the increasing prevalence of obesity and its complications worldwide. Data on genetic determinants of obesity in African populations are rare. RESEARCH METHODS AND PROCEDURES: We have undertaken a genome-wide scan for body mass index (BMI) in 182 Nigerian families that included 769 individuals. RESULTS: The prevalence of obesity was only 5%, yet polygenic heritability for BMI was in the expected range (0.46 +/- 0.07). Tandem repeat markers (402) were typed across the genome with an average map density of 9 cM. Pedigree-based analysis using a variance components linkage model demonstrated evidence for linkage on chromosome 7 (near marker D7S817 at 7p14) with a logarithm of odds (LOD) score of 3.8 and on chromosome 11 (marker D11S2000 at 11q22) with an LOD score of 3.3. Weaker evidence for linkage was found on chromosomes 1 (1q21, LOD = 2.2) and 8 (8p22, LOD = 2.3). Several candidate genes, including neuropeptide Y, DRD2, APOA4, lamin A/C, and lipoprotein lipase, lie in or close to the chromosomal regions where strong linkage signals were found. DISCUSSION: The findings of this study suggest that, as in other populations with higher prevalences of obesity, positive linkage signals can be found on genome scans for obesity-related traits. Follow-up studies may be warranted to investigate these linkages, especially the one on chromosome 11, which has been reported in a population at the opposite end of the BMI distribution. PMID- 12582224 TI - Weight goals in a college-age population. AB - OBJECTIVE: Although a growing body of literature has found unrealistic weight loss goals to be common among older, primarily female, subjects, little is known about weight loss goals of younger adults. RESEARCH METHODS AND PROCEDURES: Three hundred seventy-nine college students had their height and weight taken and reported their "goal," "dream," "happy," "acceptable," and "disappointed" weights. A series of 2 (gender) x 2 (nonoverweight vs. overweight) ANOVAs were conducted with both absolute weight goals and percentage of weight loss needed to obtain those goals as dependent variables. RESULTS: When examined in terms of absolute weight goals, women generally had lower body mass index (BMI) goals than men, and nonoverweight participants had lower BMI goals than overweight participants. Surprisingly, most overweight participants would accept a weight loss that would still place them in the overweight BMI range. When examined in terms of percentage loss needed to reach those goals, only overweight women chose goal and dream weights that would require a loss greater than can be expected from nonsurgical weight-loss treatments, and all overweight participants chose happy and acceptable weights within 15% of current weight. DISCUSSION: Participants in this study had generally reasonable weight-loss goals, and even the most extreme weight loss goals were much more moderate than those found in previous studies. These results are surprising given the extreme social pressures for thinness facing young adults. Future studies should examine the variables that influence selection of goal weights and how goal weights affect actual dieting behavior. PMID- 12582225 TI - Increased adiposity in female rhesus monkeys exposed to androgen excess during early gestation. AB - OBJECTIVE: The aim of this study was to investigate the effects of early gestational androgen excess on adult body fat distribution in female rhesus monkeys. RESEARCH METHODS AND PROCEDURES: Six midreproductive-aged, adult female rhesus monkeys that were exposed to androgen excess started during the first one third of gestation were pair-matched to control females by age, body weight, and body mass index. Body composition was determined using somatometrics, DXA, and computed tomography. RESULTS: Total abdominal and intra-abdominal fat depots are increased in adult female rhesus monkeys exposed to prenatal androgen excess. DISCUSSION: Early gestational androgen excess in female rhesus monkeys causes a preferential accumulation of total abdominal and intra-abdominal fat during adulthood. Fat accumulation in these regions is independent of total body adiposity, occurring throughout the spectrum of body mass index in these animals. This study establishes alterations in abdominal adiposity as another consequence of prenatal androgen excess in female rhesus monkeys that may contribute to the impaired insulin secretion observed in these animals during adulthood. PMID- 12582226 TI - Prescribed "breaks" as a means to disrupt weight control efforts. AB - OBJECTIVE: Research on long-term maintenance of weight loss is hampered by the fact that relapse typically does not occur until after 6 months. We sought to determine whether relapses could be induced earlier by intentionally interrupting the momentum of weight loss during a treatment program and thus provide a model for weight maintenance research. RESEARCH METHODS AND PROCEDURES: Subjects (N = 142) were recruited at two centers and randomly assigned to a control group, a long break group (LB), or a short break group (SB). The control group received a standard behavioral weight loss program with 14 weekly sessions; the LB and SB groups received the same. However, the LB group took a 6-week break after lesson 7, and the SB group took a 2-week break after lessons 3, 6, and 9. Participants were instructed to stop all weight loss efforts during breaks. RESULTS: Participants seemed to take breaks as prescribed, with interruptions in self monitoring, dietary adherence, and self-weighing (but not in exercise). However, participants quickly resumed these behaviors after the break. Breaks produced a slowing of weight loss or slight regain, but weight losses during the breaks were not significantly different from the control group. Importantly, overall weight losses (0 to 5 months or 0 to 11 months) did not differ between conditions. DISCUSSION: This study was not successful in developing a method to experimentally produce weight loss relapses. However, the finding that prescribed breaks do not have adverse effects may have clinical application. PMID- 12582228 TI - Epicardial fat from echocardiography: a new method for visceral adipose tissue prediction. AB - OBJECTIVE: To validate transthoracic echocardiography as an easy and reliable imaging method for visceral adipose tissue (VAT) prediction. VAT is recognized as an important indicator of high cardiovascular and metabolic risk. Several methods are applied to estimate VAT, with different results. RESEARCH METHODS AND PROCEDURES: We selected 60 healthy subjects (29 women, 31 men, 49.5 +/- 16.2 years) with a wide range of body mass indexes. Each subject underwent transthoracic echocardiogram and magnetic resonance imaging (MRI) to measure epicardial fat thickness on the right ventricle. Measurements of epicardial adipose tissue thickness were obtained from the same echocardiographic and MRI views and points. MRI was also used to measure VAT cross-sectional areas at the level of L4 to L5. Anthropometric indexes were also measured. RESULTS: Subjects with predominant visceral fat accumulation showed higher epicardial adipose tissue thickness than subjects with predominant peripheral fat distribution: 9.97 +/- 2.88 vs. 4.34 +/- 1.98 (p = 0.005) and 7.19 +/- 2.74 vs. 3.43 +/- 1.64 (p = 0.004) in men and women, respectively. Simple linear regression analysis showed an excellent correlation between epicardial adipose tissue and waist circumference (r = 0.895, p = 0.01) and MRI abdominal VAT (r = 0.864, p = 0.01). Multiple regression analysis showed that epicardial adipose tissue thickness (r(2) = 0.442, p = 0.02) was the strongest independent variable correlated to MRI VAT. Bland test confirmed the good agreement between the two methods. DISCUSSION: Epicardial adipose tissue showed a strong correlation with anthropometric and imaging measurements of VAT. Hence, transthoracic echocardiography could be an easy and reliable imaging method for VAT prediction. PMID- 12582227 TI - DRF 2655: a unique molecule that reduces body weight and ameliorates metabolic abnormalities. AB - OBJECTIVE: Preclinical evaluation of DRF 2655, a peroxisome proliferator activated receptor alpha (PPARalpha) and PPARgamma agonist, as a body-weight lowering, hypolipidemic and euglycemic agent. RESEARCH METHODS AND PROCEDURES: DRF 2655 was studied in different genetic, normal, and hyperlipidemic animal models. HEK 293 cells were used to conduct the reporter-based transactivation of PPARalpha and PPARgamma. To understand the biochemical mechanism of lipid-, body weight-, and glucose-lowering effects, activities of key beta-oxidation and lipid catabolism enzymes and gluconeogenic enzymes were studied in db/db mice treated with DRF 2655. 3T3L1 cells were used for adipogenesis study, and HepG2 cells were used to study the effect of DRF 2655 on total cholesterol and triglyceride synthesis using [(14)C]acetate and [(3)H]glycerol. RESULTS: DRF 2655 showed concentration-dependent transactivation of PPARalpha and PPARgamma. In the 3T3L1 cell-differentiation study, DRF 2655 and rosiglitazone showed 369% and 471% increases, respectively, in triglyceride accumulation. DRF 2655 showed body weight lowering and euglycemic and hypolipidemic effects in various animal models. db/db mice treated with DRF 2655 showed 5- and 3.6-fold inhibition in phosphoenolpyruvate carboxykinase and glucose 6-phosphatase activity and 651% and 77% increases in the beta-oxidation enzymes carnitine palmitoyltransferase and carnitine acetyltransferase, respectively. HepG2 cells treated with DRF 2655 showed significant reduction in lipid synthesis. DISCUSSION: DRF 2655 showed excellent euglycemic and hypolipidemic activities in different animal models. An exciting finding is its body-weight lowering effect in these models, which might be mediated by the induction of target enzymes involved in hepatic lipid catabolism through PPARalpha activation. PMID- 12582229 TI - Treatment with cabergoline is associated with weight loss in patients with hyperprolactinemia. PMID- 12582231 TI - Molecular evolution of eukaryotic genomes: hemiascomycetous yeast spliceosomal introns. AB - As part of the exploratory sequencing program Genolevures, visual scrutinisation and bioinformatic tools were used to detect spliceosomal introns in seven hemiascomycetous yeast species. A total of 153 putative novel introns were identified. Introns are rare in yeast nuclear genes (<5% have an intron), mainly located at the 5' end of ORFs, and not highly conserved in sequence. They all share a clear non-random vocabulary: conserved splice sites and conserved nucleotide contexts around splice sites. Homologues of metazoan snRNAs and putative homologues of SR splicing factors were identified, confirming that the spliceosomal machinery is highly conserved in eukaryotes. Several introns' features were tested as possible markers for phylogenetic analysis. We found that intron sizes vary widely within each genome, and according to the phylogenetic position of the yeast species. The evolutionary origin of spliceosomal introns was examined by analysing the degree of conservation of intron positions in homologous yeast genes. Most introns appeared to exist in the last common ancestor of present day yeast species, and then to have been differentially lost during speciation. However, in some cases, it is difficult to exclude a possible sliding event affecting a pre-existing intron or a gain of a novel intron. Taken together, our results indicate that the origin of spliceosomal introns is complex within a given genome, and that present day introns may have resulted from a dynamic flux between intron conservation, intron loss and intron gain during the evolution of hemiascomycetous yeasts. PMID- 12582232 TI - A small nuclear RNA, hdm365, is the major processing product of the human mdm2 gene. AB - mdm2 encodes for an E3 ubiquitin ligase targeting constitutively expressed p53 for proteasomal degradation. Several protein isoforms have been described for human MDM2 (HDM2), some of which may correspond to splicing variants detectable by RT-PCR in many tumors. Upon cellular stress, p53 becomes resistant to MDM2 and, in a feedback loop, up-regulates mdm2 transcription. The physiological relevance of stress-induced mdm2 gene activity is not well understood. We describe a small nuclear RNA of 365 bases comprised of the first five hdm2 exons and lacking polyadenylation. hdm365 precedes full-length hdm2 RNA expression after induction by p53 and accumulates to significant levels in the nucleus, detectable at the site of hdm2 transcription and processing only. Considering a 10-fold lower stability and high steady-state levels of the novel RNA species, hdm365 appears to be the major processing product of hdm2 transcripts. hdm365 induction was observed after ectopic expression of p53 and after DNA damaging treatment of tumor cell lines, primary fibroblasts and lymphocytes, and was not related to apoptosis. Corresponding truncated transcripts were observed in hdm2 amplified cells. High stress-inducible expression levels, absence of a corresponding protein, and nuclear localisation of hdm365 suggest a novel RNA based function for hdm2. PMID- 12582233 TI - Characterization of Arabidopsis thaliana ortholog of the human breast cancer susceptibility gene 1: AtBRCA1, strongly induced by gamma rays. AB - hBRCA1 is involved in 20-45% of inherited breast cancer cases and is implicated in many mechanisms involved in response to DNA damage. To date, BRCA1 orthologs have been characterized in vertebrate genomes only. We have identified the ortholog of BRCA1 in Arabidopsis thaliana. AtBRCA1 is a 5.5 kb part of the locus At4g21070. The corresponding mRNA of 3.5 kb is composed of 14 exons and encodes a 941 amino acid protein (104 kDa). AtBRCA1, which has one N-terminal RING finger, two C-terminal BRCT and the p300/CBP interacting domain, shows a high similarity to hBRCA1 in these motifs and has the same characteristic molecular organization. We have also identified a putative ortholog in rice (OsBRCA1). With 941 and 968 amino acids, respectively, AtBRCA1 and OsBRCA1 are the shortest members of the BRCA1 family, and may represent a plant specificity. AtBRCA1 is expressed ubiquitously in plant tissues, at levels depending on organ type, with highest levels in flower buds and exponentially growing cell cultures. Increase of mRNA levels in all plant tissues 1 h after irradiation with the highest induction level of approximately 150 times for a 100 Gy dose is consistent with a putative role of AtBRCA1 in DNA repair and in cell-cycle control. PMID- 12582234 TI - Structural transition from antiparallel to parallel G-quadruplex of d(G4T4G4) induced by Ca2+. AB - Guanine quadruplex (G-quadruplex) structures are formed by guanine-rich oligonucleotides. Because of their in vivo and in vitro importance, numerous studies have been demonstrated that the structure and stability of the G quadruplex are dependent on the sequence of oligonucleotide and environmental conditions such as existing cations. Previously, we quantitatively investigated the divalent cation effects on the antiparallel G-quadruplex of d(G4T4G4), and found that Ca2+ induces a structural transition from the antiparallel to parallel G-quadruplex, and finally G-wire formation. In the present study, we report in detail the kinetic and thermodynamic analyses of the structural transition induced by Ca2+ using stopped-flow apparatus, circular dichroism, size-exclusion chromatography (SEC) and atomic force microscopy. The quantitative parameters showed that at least two Ca2+ ions were required for the transition. The kinetic parameters also indicated that d(G4T4G4) underwent the transition through multiple steps involving the Ca2+ binding, isomerization and oligomerization of d(G4T4G4). The parallel-stranded G-wire structure of d(G4T4G4), which is a well controlled alignment of numerous DNA strands with G-quartets, as the final product induced by Ca2+, was observed using SEC and atomic force microscopy. These results provide insight into the mechanism of the structural transition and G-wire formation and are useful for constructing a nanomaterial regulated by Ca2+. PMID- 12582235 TI - Gene conversion tracts in Saccharomyces cerevisiae can be extremely short and highly directional. AB - Gene conversion is a common outcome of double-strand break (DSB) repair in yeast. Prior studies revealed that DSB-induced gene conversion tracts are often short (<53 bp), unidirectional, and biased toward promoter-proximal (5') markers. In those studies, broken ends had short, non-homologous termini. For the present study we created plasmid x chromosome, chromosomal direct repeat and allelic recombination substrates in which donor alleles carried mutant HO sites (HOinc- not cleaved) at the same position as cleavable HO sites in recipient alleles. In these substrates, broken ends are almost completely homologous to donor alleles, differing only at the three HOinc mutations. These mutations serve as markers very close to, or within, the four-base overhang produced by HO nuclease. We identified extremely short tracts (<12 bp) and many tracts were highly directional, extending <2 bp on one side of the DSB. We thought that terminal homology would promote bidirectional tracts, but found instead that unidirectional tracts were more frequent. Interestingly, substrates with terminal homology displayed enhanced 3' conversion, and in several cases conversion bias was reversed toward 3' markers. These results are discussed in relation to factors that may influence tract length and directionality, including heteroduplex DNA formation, transcription, replication and mismatch repair. PMID- 12582237 TI - Beta-globin locus control region HS2 and HS3 interact structurally and functionally. AB - The overall structure of the DNase I hypersensitive sites (HSs) that comprise the beta-globin locus control region (LCR) is highly conserved among mammals, implying that the HSs have conserved functions. However, it is not well understood how the LCR HSs, either individually or collectively, activate transcription. We analyzed the interactions of HS2, HS3 and HS4 with the human epsilon- and beta-globin genes in chromatinized episomes in fetal/embryonic K562 cells. Only HS2 activates transcription of the epsilon-globin gene, while all three HSs activate the beta-globin gene. HS3 stimulates the beta-globin gene constitutively, but HS2 and HS4 transactivation requires expression of the transcription factor EKLF, which is not present in K562 cells but is required for beta-globin expression in vivo. To begin addressing how the individual HSs may interact with one another in a complex, we linked the beta-globin gene to both the HS2 and HS3. HS2 and HS3 together resulted in synergistic stimulation of beta globin transcription. Unexpectedly, mutated, inactive forms of HS2 impeded the activation of the beta-globin gene by HS3. Thus, there appear to be distinct interactions among the HSs and between the HSs and the globin genes. These preferential, non-exclusive interactions may underlie an important structural and functional cooperativity among the regulatory sequences of the beta-globin locus in vivo. PMID- 12582236 TI - Expression of tetanus toxin Fragment C in tobacco chloroplasts. AB - Fragment C (TetC) is a non-toxic 47 kDa polypeptide fragment of tetanus toxin that can be used as a subunit vaccine against tetanus. Expression of TetC in Escherichia coli and yeast was dependent on the availability of synthetic genes that were required to improve translation efficiency and stabilize the mRNA. To explore the feasibility of producing TetC in tobacco leaves, we attempted expression of both the bacterial high-AT (72.3% AT) and the synthetic higher-GC genes (52.5% AT) in tobacco chloroplasts. We report here that the bacterial high AT mRNA is stable in tobacco chloroplasts. Significant TetC accumulation was obtained from both genes, 25 and 10% of total soluble cellular protein, respectively, proving the versatility of plastids for expression of unmodified high-AT and high-GC genes. Mucosal immunization of mice with the plastid- produced TetC induced protective levels of TetC antibodies. Thus, expression of TetC in chloroplasts provides a potential route towards the development of a safe, plant-based tetanus vaccine for nasal and oral applications. PMID- 12582238 TI - Identification of high excision capacity for 5-hydroxymethyluracil mispaired with guanine in DNA of Escherichia coli MutM, Nei and Nth DNA glycosylases. AB - The oxidation and deamination of 5-methylcytosine (5mC) in DNA generates a base pair between 5-hydroxymethyluracil (5hmU) and guanine. 5hmU normally forms a base pair with adenine. Therefore, the conversion of 5mC to 5hmU is a potential pathway for the generation of 5mC to T transitions. Mammalian cells have high levels of activity of 5hmU-DNA glycosylase, which excises 5hmU from DNA. However, glycosylases that similarly excise 5hmU have not been observed in yeast or Escherichia coli. Recently, we found that E.coli MutM, Nei and Nth have DNA glycosylase activity for 5-formyluracil, which is another type of oxidation product of the thymine methyl group. In this study, we examined whether or not E.coli MutM, Nei and Nth have also DNA glycosylase activity that acts on 5hmU in vitro. When incubated with synthetic duplex oligonucleotides containing 5hmU:G or 5hmU:A, purified MutM, Nei and Nth cleaved the 5hmU:G oligonucleotide 58, 5 and 37 times, respectively, more efficiently than the 5hmU:A oligonucleotide. In E.coli, the 5hmU-DNA glycosylase activities of MutM, Nei and Nth may play critical roles in the repair of 5hmU:G mispairs to avoid 5mC to T transitions. PMID- 12582239 TI - Translational nonsense codon suppression as indicator for functional pre-tRNA splicing in transformed Arabidopsis hypocotyl-derived calli. AB - The transient expression of three novel plant amber suppressors derived from a cloned Nicotiana tRNA(Ser)(CGA), an Arabidopsis intron-containing tRNA(Tyr)(GTA) and an Arabidopsis intron-containing tRNA(Met)(CAT) gene, respectively, was studied in a homologous plant system that utilized the Agro bacterium-mediated gene transfer to Arabidopsis hypocotyl explants. This versatile system allows the detection of beta-glucuronidase (GUS) activity by histochemical and enzymatic analyses. The activity of the suppressors was demonstrated by the ability to suppress a premature amber codon in a modified GUS gene. Co-transformation of Arabidopsis hypocotyls with the amber suppressor tRNA(Ser) gene and the GUS reporter gene resulted in approximately 10% of the GUS activity found in the same tissue transformed solely with the functional control GUS gene. Amber suppressor tRNAs derived from intron-containing tRNA(Tyr) or tRNA(Met) genes were functional in vivo only after some additional gene manipulations. The G3:C70 base pair in the acceptor stem of tRNA(Met)(CUA) had to be converted to a G3:U70 base pair, which is the major determinant for alanine tRNA identity. The inability of amber suppressor tRNA(Tyr) to show any activity in vivo predominantly results from a distorted intron secondary structure of the corresponding pre-tRNA that could be cured by a single nucleotide exchange in the intervening sequence. The improved amber suppressors tRNA(Tyr) and tRNA(Met) were subsequently employed for studying various aspects of the plant-specific mechanism of pre-tRNA splicing as well as for demonstrating the influence of intron-dependent base modifications on suppressor activity. PMID- 12582240 TI - DNA crosslinking and biological activity of a hairpin polyamide-chlorambucil conjugate. AB - A prototype of a novel class of DNA alkylating agents, which combines the DNA crosslinking moiety chlorambucil (Chl) with a sequence-selective hairpin pyrrole imidazole polyamide ImPy-beta-ImPy-gamma-ImPy-beta-Dp (polyamide 1), was evaluated for its ability to damage DNA and induce biological responses. Polyamide 1-Chl conjugate (1-Chl) alkylates and interstrand crosslinks DNA in cell-free systems. The alkylation occurs predominantly at 5'-AGCTGCA-3' sequence, which represents the polyamide binding site. Conjugate-induced lesions were first detected on DNA treated for 1 h with 0.1 micro M 1-Chl, indicating that the conjugate is at least 100-fold more potent than Chl. Prolonged incubation allowed for DNA damage detection even at 0.01 micro M concentration. Treatment with 1-Chl decreased DNA template activity in simian virus 40 (SV40) in vitro replication assays. 1-Chl inhibited mammalian cell growth, genomic DNA replication and cell cycle progression, and arrested cells in the G2/M phase. Moreover, cellular effects were observed at 1-Chl concentrations similar to those needed for DNA damage in cell-free systems. Neither of the parent compounds, unconjugated Chl or polyamide 1, demonstrated any cellular activity in the same concentration range. The conjugate molecule 1-Chl possesses the sequence-selectivity of a polyamide and the enhanced DNA reactivity of Chl. PMID- 12582241 TI - A single-nucleotide natural variation (U4 to C4) in an influenza A virus promoter exhibits a large structural change: implications for differential viral RNA synthesis by RNA-dependent RNA polymerase. AB - The influenza A virus promoter is recognized by the influenza A virus RNA dependent RNA polymerase, and directs both transcription and replication of the viral RNA genome. Within the sequence of this promoter, flu strains exhibit a natural, unique variation, either a U or a C, at the fourth position from the 3' end. Promoters that contain a C residue (C4 promoter), which are invariably found in genome segments that encode the three RNA polymerase subunits (PB1, PB2 and PA), down-regulate transcription but activate genome replication. Here, we have determined the structure of the C4 promoter by NMR spectroscopy and compared it with the structure of the U4 promoter, which was determined previously. The structure of the internal loop in the C4 promoter is similar to that of the U4 promoter. However, the terminal stem of the C4 promoter is strikingly different from that of the U4 promoter. These structural data suggest that the internal loop is important for polymerase binding to the promoter, and the terminal stem is crucial for differential regulation of transcription and replication. PMID- 12582243 TI - Evolution of transcription factors and the gene regulatory network in Escherichia coli. AB - The most detailed information presently available for an organism's transcriptional regulation network is that for the prokaryote Escherichia coli. In order to gain insight into the evolution of the E.coli regulatory network, we analysed information obtainable for the domains and protein families of the transcription factors and regulated genes. About three-quarters of the 271 transcription factors we identified are two-domain proteins, consisting of a DNA binding domain along with a regulatory domain. The regulatory domains mainly bind small molecules. Many groups of transcription factors have identical domain architectures, and this implies that roughly three-quarters of the transcription factors have arisen as a consequence of gene duplication. In contrast, there is little evidence of duplication of regulatory regions together with regulated genes or of transcription factors together with regulated genes. Thirty-eight, out of the 121 transcription factors for which one or more regulated genes are known, regulate other transcription factors. This amplification effect, as well as large differences between the numbers of genes directly regulated by transcription factors, means that there are about 10 global regulators which each control many more genes than the other transcription factors. PMID- 12582242 TI - Rap1p and other transcriptional regulators can function in defining distinct domains of gene expression. AB - Barrier elements that are able to block the propagation of transcriptional silencing in yeast are functionally similar to chromatin boundary/insulator elements in metazoans that delimit functional chromosomal domains. We show that the upstream activating sequences of many highly expressed ribosome protein genes and glycolytic genes exhibit barrier activity. Analyses of these barriers indicate that binding sites for transcriptional regulators Rap1p, Abf1p, Reb1p, Adr1p and Gcn4p may participate in barrier function. We also present evidence suggesting that Rap1p is directly involved in barrier activity, and its barrier function correlates with local changes in chromatin structure. We further demonstrate that tethering the transcriptional activation domain of Rap1p to DNA is sufficient to recapitulate barrier activity. Moreover, targeting the activation domain of Adr1p or Gcn4p also establishes a barrier to silencing. These results support the notion that transcriptional regulators could also participate in delimiting functional domains in the genome. PMID- 12582244 TI - ATIDB: Arabidopsis thaliana insertion database. AB - Insertional mutagenesis techniques, including transposon- and T-DNA-mediated mutagenesis, are key resources for systematic identification of gene function in the model plant species Arabidopsis thaliana. We have developed a database (http://atidb.cshl.org/) for archiving, searching and analyzing insertional mutagenesis lines. Flanking sequences from approximately 10 500 insertion lines (including transposon and T-DNA insertions) from several tagging programs in Arabidopsis were mapped to the genome sequence through our annotation system before being entered into the database. The database front end provides World Wide Web searching and analyzing interfaces for genome researchers and other biologists. Users can search the database to identify insertions in a particular gene or perform genome-wide analysis to study the distribution and preference of insertions. Tools integrated with the database include a graphical genome browser, a protein search function, a graphical representation of the insertion distribution and a Blast search function. The database is based on open source components and is available under an open source license. PMID- 12582245 TI - Protein-protein interaction map for yeast TFIID. AB - A major rate-limiting step in transcription initiation by RNA polymerase II is recognition and binding of the TATA element by the transcription factor TFIID. TFIID is composed of TATA binding protein (TBP) and approximately a dozen TBP associated factors (TAFs). Emerging consensus regarding the role of TAFs is that TFIID assumes a gene specific activity that is regulated by interaction with other factors. In spite of many studies demonstrating the essential nature of TAFs in transcription, very little is known about the subunit contacts within TFIID. To understand fully the functional role of TAFs, it is imperative to define TAF-TAF interactions and their topological arrangement within TFIID. We performed a systematic two-hybrid analysis using the 13 essential TAFs of the Saccharomyces cerevisiae TFIID complex and TBP. Specific interactions were defined for each component, and the biological significance of these interactions is supported by numerous genetic and biochemical studies. By combining the interaction profiles presented here, and the available studies utilizing specific TAFs, we propose a working hypothesis for the arrangement of components in the TFIID complex. Thus, these results serve as a foundation for understanding the overall architecture of yeast TFIID. PMID- 12582247 TI - A promoter activity is present in the DNA sequence corresponding to the hepatitis C virus 5' UTR. AB - The hepatitis C virus (HCV) 5' untranslated region (UTR) has been extensively studied with regard to its internal ribosomal entry site (IRES) activity. In this work we present results suggesting the existence of a strong promoter activity carried by the DNA sequence corresponding to the HCV 5' UTR. This activity was not detected when the HCV 5' UTR sequence was replaced by HCV 3' UTR or poliovirus 5' UTR sequences. These results were further confirmed by using bicistronic constructions. We demonstrated the presence of an mRNA initiated in this 5' UTR sequence and located the initiation site by the 5' RACE method at nucleotide 67. Furthermore, northern experiments and flow cytometry analysis showed the unambiguous activity of such a promoter sequence in stably transfected cells. Our results strongly suggest that the data obtained using bicistronic DNA constructs carrying the HCV 5' UTR should be analyzed not only at the translational but also at the transcriptional level. PMID- 12582246 TI - Mutations in the histone fold domain of the TAF12 gene show synthetic lethality with the TAF1 gene lacking the TAF N-terminal domain (TAND) by different mechanisms from those in the SPT15 gene encoding the TATA box-binding protein (TBP). AB - The general transcription factor TFIID, composed of the TATA box-binding protein (TBP) and 14 TBP-associated factors (TAFs), is important for both basal and regulated transcription by RNA polymerase II. Although it is well known that the TAF N-terminal domain (TAND) at the amino-terminus of the TAF1 protein binds to TBP and thereby inhibits TBP function in vitro, the physiological role of this domain remains obscure. In our previous study, we screened for mutations that cause lethality when co-expressed with the TAF1 gene lacking TAND (taf1 DeltaTAND) and identified two DeltaTAND synthetic lethal (nsl) mutations as those in the SPT15 gene encoding TBP. In this study we isolated another nsl mutation in the same screen and identified it to be a mutation in the histone fold domain (HFD) of the TAF12 gene. Several other HFD mutations of this gene also exhibit nsl phenotypes, and all of them are more or less impaired in transcriptional activation in vivo. Interestingly, a set of genes affected in the taf1-DeltaTAND mutant is similarly affected in the taf12 HFD mutants but not in the nsl mutants of TBP. Therefore, we discovered that the nsl mutations of these two genes cause lethality in the taf1-DeltaTAND mutant by different mechanisms. PMID- 12582248 TI - Inhibition of human papilloma virus E2 DNA binding protein by covalently linked polyamides. AB - Polyamides are a class of heterocyclic small molecules with the potential of controlling gene expression by binding to the minor groove of DNA in a sequence specific manner. To evaluate the feasibility of this class of compounds as antiviral therapeutics, molecules were designed to essential sequence elements occurring numerous times in the HPV genome. This sequence element is bound by a virus-encoded transcription and replication factor E2, which binds to a 12 bp recognition site as a homodimeric protein. Here, we take advantage of polyamide:DNA and E2:DNA co-crystal structural information and advances in polyamide synthetic chemistry to design tandem hairpin polyamides that are capable of displacing the major groove-binding E2 homodimer from its DNA binding site. The binding of tandem hairpin polyamides and the E2 DNA binding protein to the DNA site is mutually exclusive even though the two ligands occupy opposite faces of the DNA double helix. We show with circular permutation studies that the tandem hairpin polyamide prevents the intrinsic bending of the E2 DNA site important for binding of the protein. Taken together, these results illustrate the feasibility of inhibiting the binding of homodimeric, major groove-binding transcription factors by altering the local DNA geometry using minor groove binding tandem hairpin polyamides. PMID- 12582250 TI - Smad proteins regulate transcriptional induction of the SM22alpha gene by TGF beta. AB - Smad proteins transduce signals from transforming growth factor-beta (TGF-beta) receptors and regulate transcription of target genes. TGF-beta is implicated in the regulation of the smooth muscle cell specific gene SM22alpha, but little is known about how Smads are involved in SM22alpha gene transcription. In this report, we demonstrate that TGF-beta activation of the SM22alpha promoter is Smad dependent in C3H10T1/2 cells, BALB 3T3 cells and neural crest Monc-1 cells. We find that the promoter region from -162 to +41 is sufficient to up-regulate the reporter gene upon TGF-beta induction. Smad3, Smad1 and Smad4 are found in TGF beta inducible complexes that bind to a region containing a Smad binding site (SBS) and a medea box. Both the SBS and medea box are necessary for complex formation and are functionally important. Smad4 is limiting for TGF-beta induction, and Smad3, but not Smad1, significantly contributes to maximal activation. Time course luciferase assays and time course gel mobility shift assays reveal that the Smad3/4 complex is largely responsible for the immediate response of the SM22alpha promoter to TGF-beta induction, and also contributes to the maximal promoter activity. We further demonstrate that AP-1 elements contribute to induction of the SM22alpha promoter by TGF-beta. PMID- 12582249 TI - Characterization of the human beta-globin downstream promoter region. AB - The human beta-globin gene is abundantly expressed specifically in adult erythroid cells. Stage-specific transcription is regulated principally by promoter proximal cis-regulatory elements. The basal promoter contains a non canonical TATA-like motif as well as an initiator element. These two elements have been shown to interact with the TFII-D complex. Here we show that in addition to the TATA and initiator elements, conserved E-box motifs are located in the beta-globin downstream promoter. One of the E-box motifs overlaps the initiator and this composite element interacts with USF1 and TFII-I in vitro. Another E-box, located 60 bp 3' to the transcription initiation site, interacts with USF1 and USF2. Mutations of either the initiator or the downstream E-box impair transcription of the beta-globin gene in vitro. Mutations of a putative NF E2-binding site in the downstream promoter region do not affect transcription in vitro. USF1, USF2, TFII-I and p45 can be crosslinked to a beta-globin promoter fragment in MEL cells in vivo, whereas only TFII-I and USF2 crosslink to the beta globin gene in K562 cells. The summary data demonstrate that in addition to the well-characterized interactions of the TFII-D complex with the basal promoter, E box motifs contribute to the efficient formation of transcription complexes on the adult beta-globin gene. PMID- 12582251 TI - Discrimination among individual Watson-Crick base pairs at the termini of single DNA hairpin molecules. AB - Nanoscale alpha-hemolysin pores can be used to analyze individual DNA or RNA molecules. Serial examination of hundreds to thousands of molecules per minute is possible using ionic current impedance as the measured property. In a recent report, we showed that a nanopore device coupled with machine learning algorithms could automatically discriminate among the four combinations of Watson-Crick base pairs and their orientations at the ends of individual DNA hairpin molecules. Here we use kinetic analysis to demonstrate that ionic current signatures caused by these hairpin molecules depend on the number of hydrogen bonds within the terminal base pair, stacking between the terminal base pair and its nearest neighbor, and 5' versus 3' orientation of the terminal bases independent of their nearest neighbors. This report constitutes evidence that single Watson-Crick base pairs can be identified within individual unmodified DNA hairpin molecules based on their dynamic behavior in a nanoscale pore. PMID- 12582252 TI - Hybridization kinetics and thermodynamics of molecular beacons. AB - Molecular beacons are increasingly being used in many applications involving nucleic acid detection and quantification. The stem-loop structure of molecular beacons provides a competing reaction for probe-target hybridization that serves to increase probe specificity, which is particularly useful when single-base discrimination is desired. To fully realize the potential of molecular beacons, it is necessary to optimize their structure. Here we report a systematic study of the thermodynamic and kinetic parameters that describe the molecular beacon structure-function relationship. Both probe and stem lengths are shown to have a significant impact on the binding specificity and hybridization kinetic rates of molecular beacons. Specifically, molecular beacons with longer stem lengths have an improved ability to discriminate between targets over a broader range of temperatures. However, this is accompanied by a decrease in the rate of molecular beacon-target hybridization. Molecular beacons with longer probe lengths tend to have lower dissociation constants, increased kinetic rate constants, and decreased specificity. Molecular beacons with very short stems have a lower signal-to-background ratio than molecular beacons with longer stems. These features have significant implications for the design of molecular beacons for various applications. PMID- 12582253 TI - Structural effect of the anticancer agent 6-thioguanine on duplex DNA. AB - The incorporation of 6-thioguanine (S6G) into DNA is an essential step in the cytotoxic activity of thiopurines. However, the structural effects of this substitution on duplex DNA have not been fully characterized. Here, we present the solution structures of DNA duplexes containing S6G opposite thymine (S6G.T) and opposite cytosine (S6G.C), solved by high-resolution NMR spectroscopy and restrained molecular dynamics. The data indicate that both duplexes adopt right handed helical conformations with all Watson-Crick hydrogen bonding in place. The S6G.T structures exhibit a wobble-type base pairing at the lesion site, with thymine shifted toward the major groove and S6G displaced toward the minor groove. Aside from the lesion site, the helices, including the flanking base pairs, are not highly perturbed by the presence of the lesion. Surprisingly, thermal dependence experiments suggest greater stability in the S6G-T mismatch than the S6G-C base pair. PMID- 12582254 TI - Structural divergence of chromosomal segments that arose from successive duplication events in the Arabidopsis genome. AB - Using the extensive segmental duplications of the Arabidopsis thaliana genome, a comparative study of homoeologous segments occurring in chromosomes 1, 2, 4 and 5 was performed. The gene-by-gene BLASTP approach was applied to identify duplicated genes in homoeologues. The levels of synonymous substitutions between duplicated coding sequences suggest that these regions were formed by at least two rounds of duplications. Moreover, remnants of even more ancient duplication events were recognised by a whole-genome study. We describe a subchromosomal organisation of genes, including the tandemly repeated genes, and the distribution of transposable elements (TEs). In certain cases, evidence of the possible mechanisms of structural rearrangements within the segments could be found. We provide a probable scenario of the rearrangements that took place during the evolution of the homoeologous regions. Furthermore, on the basis of the comparative analysis of the chromosomal segments in the Columbia and Landsberg erecta accessions, an additional structural variation in the A.thaliana genome is described. Analysis of the segments, spanning 7 Mb or 5.6% of the genome, permitted us to propose a model of evolution at the subchromosomal level. PMID- 12582255 TI - Limited repair of 8-hydroxy-7,8-dihydroguanine residues in human testicular cells. AB - Oxidative damage in testicular DNA is associated with poor semen quality, reduced fertility and increased risk of stillbirths and birth defects. These DNA lesions are predominantly removed by base excision repair. Cellular extracts from human and rat testicular cells and three enriched populations of rat male germ cells (primary spermatocytes, round spermatids and elongating/elongated spermatids) all showed proficient excision/incision of 5-hydroxycytosine, thymine glycol and 2,6 diamino-4-hydroxy-5-formamidopyrimidine. DNA containing 8-oxo-7,8-dihydroguanine was excised poorly by human testicular cell extracts, although 8-oxoguanine-DNA glycosylase-1 (hOGG1) was present in human testicular cells, at levels that varied markedly between 13 individuals. This excision was as low as with human mononuclear blood cell extracts. The level of endonuclease III homologue-1 (NTH1), which excises oxidised pyrimidines, was higher in testicular than in somatic cells of both species. Cellular repair studies of lesions recognised by formamidopyrimidine-DNA glycosylase (Fpg) or endonuclease III (Nth) were assayed with alkaline elution and the Comet assay. Consistent with the enzymatic activities, human testicular cells showed poor removal of Fpg-sensitive lesions but efficient repair of Nth-sensitive lesions. Rat testicular cells efficiently repaired both Fpg- and Nth-sensitive lesions. In conclusion, human testicular cells have limited capacity to repair important oxidative DNA lesions, which could lead to impaired reproduction and de novo mutations. PMID- 12582256 TI - A Ca2+-induced mitochondrial permeability transition causes complete release of rat liver endonuclease G activity from its exclusive location within the mitochondrial intermembrane space. Identification of a novel endo-exonuclease activity residing within the mitochondrial matrix. AB - Endonuclease G, a protein historically thought to be involved in mitochondrial DNA (mtDNA) replication, repair, recombination and degradation, has recently been reported to be involved in nuclear DNA degradation during the apoptotic process. As a result, its involvement in mtDNA homeostasis has been called into question and has necessitated detailed analyses of its precise location within the mitochondrion. Data is presented localizing rat liver endonuclease G activity exclusively to the mitochondrial intermembrane space with no activity associated with either the interior face of the inner mitochondrial membrane or with the mitochondrial matrix. Additionally, it is shown that endonuclease G can be selectively released from the mitochondrion via induction of a Ca2+-induced mitochondrial permeability transition and that, upon its release, a further nuclease activity loosely associated with the interior face of the inner mitochondrial membrane and distinct in its properties from that of endonuclease G can be detected. PMID- 12582257 TI - Rapid generation of inducible mouse mutants. AB - We have generated an optimized inducible recombination system for conditional gene targeting based on a Cre recombinase-steroid receptor fusion. This configuration allows efficient Cre-mediated recombination in most organs of the mouse upon induction, without detectable background activity. An ES cell line, was established that carries the inducible recombinase and a loxP-flanked lacZ reporter gene. Out of this line, completely ES cell-derived mice were efficiently produced through tetraploid blastocyst complementation, without the requirement of mouse breeding. Our findings provide a new concept allowing the generation of inducible mouse mutants within 6 months, as compared to 14 months using the current protocol. PMID- 12582258 TI - Directly labeled mRNA produces highly precise and unbiased differential gene expression data. AB - Microarray based gene expression studies allow simultaneous analysis of relative amounts of messenger RNA (mRNA) for thousands of genes using fluorescently labeled nucleic acid targets. Most common methods use enzymatic techniques, such as oligo-dT primed reverse transcription to produce labeled cDNA. These labeling methods have a number of shortcomings, including enzyme- introduced labeling and sequence bias, laborious protocols, high experiment-to-experiment variability and an inability to detect small changes in expression levels. Here, we describe a novel labeling methodology that uses platinum-linked cyanine dyes to directly chemically label mRNA from as little as 2 micro g of total RNA. We show that the gene expression data produced using the labeled mRNA method has very high precision, low error, no labeling bias and a dynamic range over several orders of magnitude. This allows a greater accuracy in the identification of differentially expressed genes and cuts down on the need for running too many replicate assays. Small changes in gene expression can now be detected in large-scale gene expression profiling assays using this simple, easy and quick procedure. PMID- 12582259 TI - Three color cDNA microarrays: quantitative assessment through the use of fluorescein-labeled probes. AB - Gene expression studies using microarrays have great potential to generate new insights into human disease pathogenesis, but data quality remains a major obstacle. In particular, there does not exist a method to determine prior to hybridization whether an array will yield high quality data, given good study design and target preparation. We have solved this problem through development of a three-color cDNA microarray platform where printed probes are fluorescein labeled, but are spectrally compatible with Cy3 and Cy5 dye-labeled targets when using confocal laser scanners possessing narrow bandwidths. This approach enables prehybridization evaluation of array/spot morphology, DNA deposition and retention and background levels. By using these measurements and the intra-slide coefficient of variation for fluorescence intensity we show that slides in the same batch are not equivalent and measurable prehybridization parameters can be predictive of hybridization performance as determined by replicate consistency. When hybridizing target derived from two cell lines to high and low quality replicate pairs (n = 50 pairs), a direct and significant relationship between prehybridization signal-to-background noise and post-hybridization reproducibility (R2 = 0.80, P < 0.001) was observed. We therefore conclude that slide selection based upon prehybridization quality scores will greatly benefit the ability to generate reliable gene expression data. PMID- 12582260 TI - Summaries of Affymetrix GeneChip probe level data. AB - High density oligonucleotide array technology is widely used in many areas of biomedical research for quantitative and highly parallel measurements of gene expression. Affymetrix GeneChip arrays are the most popular. In this technology each gene is typically represented by a set of 11-20 pairs of probes. In order to obtain expression measures it is necessary to summarize the probe level data. Using two extensive spike-in studies and a dilution study, we developed a set of tools for assessing the effectiveness of expression measures. We found that the performance of the current version of the default expression measure provided by Affymetrix Microarray Suite can be significantly improved by the use of probe level summaries derived from empirically motivated statistical models. In particular, improvements in the ability to detect differentially expressed genes are demonstrated. PMID- 12582262 TI - Detection and analysis of spliced chimeric mRNAs in sequence databanks. AB - We have developed a databank screening procedure, the In Silico Trans-splicing Retrieval System (ISTReS), to identify heterologous, spliced mRNAs with potential origin from chromosomal translocations, mRNA trans-splicing and multi-locus transcription. A parsing algorithm to screen cDNA versus genome Blast outputs was implemented. Key filtering criteria were Blast scores of > or =300, match lengths of > or =95% of the query sequences, junction of the two partners at exon-exon borders and concordant 'sense/sense' reading orientation. ISTReS was validated by the successful identification of bona fide chromosomal translocation-derived fusion transcripts in the HGI and RefSeq databanks. The performance of ISTReS was verified against recently identified chimeric antisense transcripts, where it revealed essentially no independent proof of antisense transcription and absence of exon-exon borders at the chimeric join, consistent with an artefactual origin. Analysis of the UNIGENE database revealed 21 742 chimeric sequences overall that correspond to approximately 1% of the database transcripts. Novel FOP-Rho GAP and methionyl tRNA synthetase-advillin chimeric mRNAs with the canonical features of heterologous-genes spliced-transcripts were identified among 246 chimeras from the RefSeq databank. This suggests a frequency of canonically-spliced chimeras of approximately 1% of all the hybrid sequences in current databanks. These findings demonstrate the efficiency of ISTReS and the overall feasibility of sequence/structure-based strategies to search for chimeric mRNAs candidate to derive from the splicing of heterologous transcripts. PMID- 12582261 TI - A yeast-based growth assay for the analysis of site-specific proteases. AB - Many cellular and viral processes depend on site-specific proteolysis. Here, a genetic system for the identification of such proteases and inhibitors is described. The system utilizes the temperature- sensitive Saccharomyces cerevisiae CDC25-2 mutant strain and exploits the strict requirement of membrane localization of a constitutively active Ras mutant for the complementation of the yeast growth defect at the non-permissive temperature. Expression of a fusion protein in which a substrate peptide of the TEV protease separates a myristoylation signal from a constitutively active human Ras protein confers temperature insensitivity. Co-expression of the protease results in release of the Ras mutant from the membrane and growth arrest at the non-permissive temperature. This non-transcriptional assay represents a new approach to the in vivo analysis of site-specific proteases and may be a valuable alternative to existing methods. It has significant potential for the selection of inhibitors of cytoplasmic and membrane-associated proteases of biotechnical and clinical relevance. PMID- 12582264 TI - Organizers' introduction to the Symposium on Quality Health Care. PMID- 12582265 TI - What is the quality of quality of medical care measures? Rashomon-like relativism and real-world applications. AB - Much attention has been directed toward the measurement of health outcomes and the quality of medical care. Some policymakers tout outcomes measurement as a promising tool for improving health care, while others question whether current quality of care assessment is valid and of practical importance. Although significant advances have been made in the ability to measure quality of care with validity, several major methodological challenges remain. We analyze the quality of quality of medical care measures, using a conceptual framework that outlines the purpose of the measures; paradigms of the quality of care, including Donabedian's structure-process-outcome model and continuous quality improvement; and other key elements, such as scope, time, unit of analysis, and perspectives. Patient preferences and organizational contexts have been underemphasized, and should be incorporated into the framework for conceptualizing quality of care. Quality of care has relativistic and dynamic aspects. Different perspectives lead to different views of what high quality care is, and quality of care is a changing concept since medical science and treatment are constantly evolving. The most appropriate quality measures depend upon whose perspective one takes and the purpose of the measures; if the measures are chosen wisely, the current state of quality of medical care measurement is adequate for both accountability and quality improvement. PMID- 12582263 TI - Identification and removal of contaminating fluorescence from commercial and in house printed DNA microarrays. AB - Microarray analysis is a critically important technology for genome-enabled biology, therefore it is essential that the data obtained be reliable. Current software and normalization techniques for microarray analysis rely on the assumption that fluorescent background within spots is essentially the same throughout the glass slide and can be measured by fluorescence surrounding the spots. This assumption is not valid if background fluorescence is spot-localized. Inaccurate estimates of background fluorescence under the spot create a source of error, especially for low expressed genes. We have identified spot-localized, contaminating fluorescence in the Cy3 channel on several commercial and in-house printed microarray slides. We determined through mock hybridizations (without labeled target) that pre-hybridization scans could not be used to predict the contribution of this contaminating fluorescence after hybridization because the change in spot-to-spot fluorescence after hybridization was too variable. Two solutions to this problem were identified. First, allowing 4 h of exposure to air prior to printing on to Corning UltraGAPS slides significantly reduced contaminating fluorescence intensities to approximately the value of the surrounding glass. Alternatively, application of a novel, hyperspectral imaging scanner and multivariate curve resolution algorithms, allowed the spectral contributions of Cy3 signal, glass, and contaminating fluorescence to be distinguished and quantified after hybridization. PMID- 12582267 TI - Indirect restraints on the provision of heath care quality. PMID- 12582268 TI - Markets and medicine: barriers to creating a "business case for quality". AB - Systematically improving the quality of medical care requires the creation and implementation of organized processes by health plans, hospitals, and physician groups. But to a considerable extent the medical market in the United States financially penalizes organizations that invest in improving quality, rather than rewarding them. This article explores the ways in which the market as presently constituted fails to reward investments in quality improvement and describes efforts newly underway to create a "business case for quality." It briefly suggests measures to that could be taken by public and private policymakers--by government as purchaser and regulator of medical care, and by large employers who in effect make policy through their health insurance purchasing decisions--to create a business case for quality. PMID- 12582270 TI - Does medicare care about quality? PMID- 12582271 TI - Health care quality, geographic variations, and the challenge of supply-sensitive care. PMID- 12582272 TI - Rethinking adaptation: the niche-construction perspective. AB - Niche construction refers to the capacity of organisms to construct, modify, and select important components of their local environments, such as nests, burrows, pupal cases, chemicals, and nutrients. A small but increasing number of evolutionary biologists regard niche construction as an evolutionary process in its own right, rather than as a mere product of natural selection. Through niche construction organisms not only influence the nature of their world, but also in part determine the selection pressures to which they and their descendants are exposed, and they do so in a non-random manner. Mathematical population genetics analyses have revealed that niche construction is likely to be evolutionarily consequential because of the feedback that it generates in the evolutionary process. A parallel movement has emerged in ecosystem ecology, where researchers stress the utility of regarding organisms as ecosystem engineers, who partly control energy and matter flows. From the niche construction standpoint, the evolving complementary match between organisms and environments is the product of reciprocal interacting processes of natural selection and niche construction. This essay reviews the arguments put forward in favor of the niche-construction perspective. PMID- 12582273 TI - Autonomy reconfigured: incorporating the role of the unconscious. AB - The principle of respect for autonomy has served as a pillar of American bioethics. Through its application in response to physician paternalism and as the basis for informed consent, it has attained preeminent status in the discipline. New challenges in health care warrant a re-examination of the origins of our autonomy. Developmental psychology, cognitive neuroscience, and behavioral neurology all emphasize the intimate relationship between our cognitive and emotional development, and through this, our conscious and unconscious minds. Our intellectual, and particularly our moral development, are instilled in us by our moral mentors, and consequently by our community as a whole. Autonomy and community are therefore inextricably interlinked, and our emphasis on liberty, rights, and privacy without the inclusion of family, community, and social responsibility is biologically unsound. Appreciation of this reality should lead to a more balanced set of bioethical norms. PMID- 12582275 TI - Hands up--it's the hygiene police! PMID- 12582274 TI - Science matters, culture matters. PMID- 12582276 TI - Speaking volumes on volunteering. PMID- 12582283 TI - The difference. PMID- 12582282 TI - What you need to know about colorectal cancer. PMID- 12582284 TI - Angina. PMID- 12582285 TI - Managing pediatric emergencies: no small matter. PMID- 12582286 TI - Protect your future with personal liability insurance. PMID- 12582287 TI - Assessing respiratory status. PMID- 12582288 TI - New drugs 2003, part I. PMID- 12582289 TI - Hypoglycemia. PMID- 12582290 TI - Great expectations. PMID- 12582291 TI - Menetrier's disease: case study in the quality of life. AB - Menetrier's disease is a type of hypertrophic gastropathy, a rare, acquired premalignant disorder of the stomach accompanied by debilitating symptoms such as nausea and vomiting, anorexia, edema, epigastric pain, and weight loss. The disease results in a marked negative effect on the patient's quality of life. This article will present information on Menetrier's disease, quality of life, and the effect this disease and subsequent treatment had on a patient's physical, emotional, and spiritual well-being. PMID- 12582292 TI - A clinical pathway for pediatric gastroenteritis. AB - Gastroenteritis is one of the most common reasons for hospitalization in the United States for children under the age of 5 years. Second only to respiratory problems, the cost of providing care for these children is estimated to exceed $2 billion dollars annually ( Burkhart, 1999; Prescilla, 2002). This article reviews the causes of gastroenteritis as well as the 1996 American Academy of Pediatrics Guidelines for treatment of gastroenteritis. These guidelines and more recently published literature were used to develop a clinical pathway to improve the care of pediatric patients admitted to the hospital with gastroenteritis. The article discusses the importance of clinical pathways and the process of implementation of a pathway for pediatric gastroenteritis. Additionally, the article provides a parent teaching tool to address recurrent questions parents have related to home management of a child with gastroenteritis. Useful web sites for resource information related to gastroenteritis are provided as well. PMID- 12582293 TI - Preoperative education improves quality of patient care for endoscopic retrograde cholangiopancreatography. AB - The goal of this study was to evaluate the effect of preoperative education on patient satisfaction and cooperation during an endoscopic retrograde cholangiopancreatography (ERCP) procedure. Risk reduction was also studied in terms of decreased meperidine administration. Subjects were preoperatively educated by the endoscopy nurse regarding the ERCP procedure and how to communicate with the nurse during the procedure. Forty-five patients with a mean age of 58 years participated in the study. Effective patient cooperation was achieved. All subjects were satisfied with the educational process and the ERCP team. These findings suggest preoperative education is a cost-effective intervention in enhance patient cooperation and patient satisfaction for patients undergoing ERCP. PMID- 12582294 TI - Palliative care update for gastroenterology nurses. AB - Changing demographics, specifically an aging population associated with an increase of chronic and terminal illness, have highlighted the need for palliative or comfort care in nursing from the time of diagnosis through the dying process. Palliative or comfort care is now on the national healthcare agenda. The purpose of this article is to provide an update on selected palliative care and end-of-life issues and resources related to practice, education, research, and policy for gastroenterology nurses. In addition, selecting one of the suggested resources to review on a regular basis will provide the reader with a strategy for keeping up-to-date in palliative and end of-life care. PMID- 12582296 TI - Capsule endoscopy: a case study of an 11-year-old girl. AB - Small-bowel disorders have typically been challenging to diagnose because of difficulty visualizing this organ. The recent development of an innovative M2A trade mark Capsule that provides video visualization of the small bowel may have a unique niche in the diagnosis of small bowel disorders. This article highlights a pediatric patient's experience of capsule endoscopy to help the reader more clearly understand the procedure and appropriate nursing care. PMID- 12582295 TI - Patient and physician evaluation of a new bulk fiber laxative tablet. AB - Occasional constipation is a frequent complaint in medical practice. Bulk-fiber laxatives are often recommended as a safe and effective treatment of this disorder. The viscous suspensions required for administering most bulk-fiber laxatives, however, can negatively affect compliance. This study summarizes physician and patient assessment of a new, oral bulk-fiber laxative caplet following 2 weeks of open-label use for the treatment of constipation and related disorders. Forty-two patients recruited from colorectal surgical practices and who were recommended bulk-fiber as part of their treatment completed the study. Recommended dosage was 2 caplets b.i.d., with each 2-caplet dose providing 1 gram of methylcellulose. Patient diaries reported average daily use of 3.13 +/- 1.20 caplets/day and a total of 577 patient-days of use (calculated as total days product used x number of subjects). Ninety percent of the diaries reported at least one bowel movement daily. Fourteen days of treatment progressively and significantly increased the percentage of complete bowel movements, bowel movements having normal stool consistency, and reduced straining. Physicians rated more than 75% of patients' responses to treatment with methylcellulose caplets as "very good" or "excellent," and considered the responses equal or better than that expected for a fiber treatment in 90% of subjects. Among 24 patients who had previously used fiber, 21 preferred the caplets over prior fiber products. Methylcellulose tablets may provide a useful alternative for improving compliance among patients using fiber therapy. PMID- 12582297 TI - Thoughts on complementary approaches to intestinal disease. PMID- 12582298 TI - In a position to influence care. PMID- 12582299 TI - The literature review. PMID- 12582300 TI - Remembering our purpose. PMID- 12582301 TI - A new policy on letters to the editor. PMID- 12582302 TI - The clinical relevance of microbiology in acute and chronic wounds. AB - PURPOSE: To improve clinical practice and the quality of patient care by providing a learning opportunity that enhances the participant's understanding of how wound microbiology affects healing. TARGET AUDIENCE: This CME/CE activity is intended for physicians and nurses with an interest in interpreting the role of microorganisms in wound healing. OBJECTIVES: At the conclusion of this course, participants should be able to: 1. Identify the microbiology of acute and chronic wounds, risk factors for infection, and advantages and disadvantages of wound culturing. 2. Identify methods of debridement and wounds for which they are appropriate. 3. Identify systemic antibiotic treatment options for acute and chronic wound infections. PMID- 12582303 TI - Pressure ulcer education: a pilot study of the knowledge and clinical confidence of geriatric fellows. AB - OBJECTIVE: To assess the educational experiences of geriatric fellows on the subject of pressure ulcers and to begin validation of a survey that assesses pressure ulcer content knowledge of physicians in training. DESIGN: Pilot study using a prospective survey. PARTICIPANTS: 42 geriatric fellows in New York State. MAIN OUTCOME MEASURES: Likert scale ranking for self-reported feeling of preparation to manage geriatric patients with pressure ulcers and to teach other clinicians about the subject; knowledge test scores using a multiple choice, single-best-answer format. RESULTS: Geriatric fellows in New York State who participated in the survey felt "adequately" prepared to lead a team and teach about pressure ulcers. Sixty-nine percent reported having teaching responsibilities. Sources of information included bedside rounds (79%), nurses (71%) lectures (67%), textbooks (67%), and geriatric attendings (60%). Educational settings were nursing homes (86%) and hospital units (64%). Forty eight percent of geriatric fellows surveyed correctly identified the Braden Scale as a screening tool. Sixty-seven percent identified a description of a Stage I pressure ulcer and 52% identified a description of a Stage IV pressure ulcer. The mean score on the knowledge test for the cohort was 58 +/- 18% (SD) correct (range, 20% to 80%); the range for the fellowship programs was 36% to 62% correct. CONCLUSIONS: Geriatric fellows need to improve their knowledge and confidence with regard to pressure ulcer care to become competent as clinicians and educators for this condition. Specific curricular guidelines and a validated knowledge assessment instrument on pressure ulcers are needed to improve the educational effectiveness of a geriatrics fellowship. PMID- 12582304 TI - Wounds and malignancy. AB - Due to the prevalence of skin cancers, health care practitioners involved with wound management are likely to encounter cutaneous malignancies as part of their practice. This article focuses on 2 ways in which malignancies and wounds are related: the malignant degeneration of chronic wounds into cancer and malignancies that present as chronic wounds. The most common scenario in which chronic wounds have been associated with the development of squamous cell carcinoma is in the presence of chronic osteomyelitis. However, wounds secondary to burns, trauma, radiotherapy, and diabetes are also at risk for malignant degeneration. It is often difficult to distinguish malignant transformations from primary malignant ulcers. Given the uncommon nature of degeneration of a chronic wound or a malignancy presenting as a chronic wound, some suggest that only suspicious wounds undergo biopsy. Primary malignancy should be considered if the ulcer has a relatively short duration and the patient does not have a history of prior radiotherapy. Until recently, amputation has been the treatment of choice for squamous cell carcinomas that arose within chronic wounds associated with chronic osteomyelitis; however, other reports have shown that other methods of ensuring complete local excision are also useful. PMID- 12582305 TI - Medicare payment issues for physicians specializing in skin and wound management. PMID- 12582306 TI - Obesity and wound healing. PMID- 12582307 TI - Nasal polyposis: an update: editorial review. AB - PURPOSE OF REVIEW: Nasal polyposis is a chronic inflammatory disease of the upper airway characterized histologically by the infiltration of inflammatory cells like eosinophils or neutrophils. Several hypotheses have been put forward regarding the underlying mechanisms including chronic infection, aspirin intolerance, alteration in aerodynamics with trapping of pollutants, epithelial disruptions, epithelial cell defects/gene deletions (CFTR gene), inhalant or food allergies. The present review is an update on the pathomechanisms of nasal polyposis. RECENT FINDINGS: In the majority of nasal polyps, eosinophils comprise more than 60% of the cell population. Besides eosinophils, mast cells and activated T cells are also increased. An increased production of cytokines/chemokines like granulocyte/macrophage colony-stimulating factor, IL-5, RANTES and eotaxin contribute to eosinophil migration and survival. Increased levels of IL-8 can induce neutrophil infiltration. Increased expression of vascular endothelial growth factor and its upregulation by transforming growth factor-beta can contribute to the edema and increased angiogenesis in nasal polyps. Again, transforming growth factor-beta can modulate fibroblast function and thus contribute to eosinophil infiltration and stromal fibrosis. Other mediators like albumin, histamine and immunoglobulins IgE and IgG are also increased in nasal polyps. In addition, the local production of IgE in nasal polyps can contribute to the increased recurrence of nasal polyps via the IgE mast cell-FcepsilonRI cascade. Finally, mast cell/T cell-epithelial cell/fibroblast interactions can contribute to the persistent eosinophilic inflammation seen in polyps. SUMMARY: Thus although nasal polyposis is a multifactorial disease with several different etiological factors, chronic persistent inflammation is undoubtedly a major factor irrespective of the etiology. PMID- 12582308 TI - Early pet exposure: friend or foe? AB - PURPOSE OF REVIEW: Sensitization to pets is a risk factor for asthma and it was assumed that pet ownership was a risk factor for sensitization. Epidemiological studies failed to confirm this, some even suggesting that keeping pets decreased the risk of sensitization and asthma. In the last year, 10 studies have been published which have, at least in part, attempted to address the question. The results, however, are heterogeneous and it is still not clear how we should advise our patients on this issue of pet ownership. RECENT FINDINGS: Results of studies of the association between exposure to cat or cat allergen and the development of sensitization are such that almost any view on the relationship could be supported by evidence from the literature. For dogs, there are fewer data, but there is little to suggest that keeping a dog increases the risk of sensitization to dog. The majority of studies reviewed find either no association or a reduced risk of asthma amongst pet owners, but only one of these selectively excludes those who deliberately avoid pets from the analysis. There is evidence to suggest that amongst non-pet owners, the risk of sensitization and of asthma increases in areas with a high proportion of pet owners. There is evidence emerging that the effect of exposure to pets may be different in different relative risk groups, based on parental allergy. There is also evidence that asthma is more severe amongst pet sensitized pet owners. SUMMARY: There are several large birth cohort studies being conducted around the world designed to measure the development of asthma and allergies in children with prospective and objective measures of environmental exposures. The results of such studies are required before the association between pets and asthma can be determined. PMID- 12582309 TI - Role of intestinal flora in the development of allergy. AB - PURPOSE OF REVIEW: The frequency of allergic diseases is increasing worldwide. Experimental and clinical studies have linked a reduced number of early infections to this trend. The gastrointestinal system, which comprises the largest lymphoid tissue and microbial reservoir of the body, has received more attention during the last few years as a potential determiner in the development of atopic disease. RECENT FINDINGS: Alterations in intestinal microbiota have been detected both in infants suffering from allergic disease and in those later developing the disorder. Delay in the compositional development of and in gut microflora was a general finding in allergic children. In a subsequent study, perinatal administration of lactobacilli halved the later development of atopic eczema during the first 2 years of life. Specific strains of the healthy gut microbiota have been shown to induce the production of IL-10 and transforming growth factor-beta, which possess an important regulative role in the development of allergic type immune response. Probiotics also strengthen gut defence barrier mechanisms and reduce antigen load in the gut. Pattern recognition receptors in intestinal epithelial and antigen-presenting cells have been demonstrated to mediate a continuing dialogue between host and gut microbiota. SUMMARY: Despite several promising findings, the exact role of gut normal microbiota in the development of allergy remains to be elucidated. For successful interventions, more data concerning a communication between host and specific microbial species are needed. PMID- 12582310 TI - Novel approaches in the treatment of allergic rhinitis. AB - PURPOSE OF REVIEW: The increasing prevalence of allergic rhinitis, its impact on individual quality of life and social costs, as well as its role as a risk factor for asthma, underline the need for improved treatment options for this disorder. We reviewed the recent literature published in English, dealing with relevant and appropriately controlled clinical trials on the treatment of allergic rhinitis. RECENT FINDINGS: In addition to well established pharmacological therapies with antihistamines, corticosteroids, decongestants and mast cell stabilizers, new therapeutic options become increasingly important, including leukotriene modifiers, anti-immunoglobulin E antibodies, phosphodiesterase inhibitors and intranasal heparin; there also exist new developments in appropriate allergen specific immunotherapy. SUMMARY: Promising results, both with established and novel treatment options, are communicated, but further studies are still necessary to confirm these data. PMID- 12582311 TI - The effect of treatment of allergic rhinitis on asthma morbidity, including emergency department visits. AB - PURPOSE OF REVIEW: It has been hypothesized that asthma and allergic rhinitis are both manifestations of a single inflammatory process present throughout the airway and that control of the inflammatory response associated with allergic rhinitis may help to reduce inflammation throughout the airway and improve control of asthma. Although previous evidence has documented that therapy directed at allergic rhinitis can improve asthma symptoms and bronchial responsiveness, until recently, there was no evidence that the use of medication targeted at the upper airway can also reduce utilization of acute health care services due to asthma. RECENT FINDINGS: We identified studies published between August 2001 and July 2002 that examined the therapeutic link between allergic rhinitis and asthma. In particular, we concentrated on investigations that evaluated whether, among patients diagnosed with allergic rhinitis and asthma, treatment of the former resulted in significantly fewer asthma-related emergency department visits or inpatient hospitalizations. Two such studies were identified. Using observational study designs, both investigations established that among a population with asthma and allergic rhinitis, treatment for allergic rhinitis was associated with a decrease in the risk of subsequent asthma-related events by one-third to one-half compared with persons who did not receive treatment for this disorder. SUMMARY: Effective management of allergic rhinitis with nasal steroids and possibly oral antihistamines reduces utilization of acute health care services due to asthma among persons with co-existent asthma. PMID- 12582312 TI - Sinusitis update. AB - PURPOSE OF REVIEW: Rhinosinusitis affects more than 10% of the US population. It results in high health care expenditure as well as decreased productivity through lost days of school or work. This review includes recent findings in diagnostics and pathophysiology, as well as recent advances in treatment. RECENT FINDINGS: Parameters used in the diagnosis of rhinosinusitis include patient symptomatology, physical and endoscopic findings, computed tomography scan scores, and culture results. There has been poor correlation between patient symptoms and computed tomography scan scores, but middle meatal cultures have been shown to correlate well with ethmoid sinus cultures obtained from chronic rhinosinusitis patients. Chronic inflammation with a predominance of eosinophils and T lymphocytes in the tissues is being recognized as a hallmark of chronic rhinosinusitis, especially in patients with asthma or allergic rhinitis. Th2 cytokines are prominent in sinus tissues of patients with chronic rhinosinusitis and allergic rhinitis. While antibiotics are used with greater frequency in the treatment of acute and chronic rhinosinusitis, adjuvant therapies, such as intranasal steroids, might affect patient symptoms and curb disease recurrence. SUMMARY: Clinical symptoms are important in making the diagnosis of rhinosinusitis and should be supported by objective findings on nasal endoscopy and, if necessary, computed tomography scans. The mainstay of treatment remains antibiotics, with a potential role for decongestants and intranasal steroids. The presence of chronic inflammation with a Th2 cytokine predominance in sinus tissues should be kept in mind, especially in patients with coexistent morbidities such as allergic rhinitis and asthma. PMID- 12582313 TI - The role of respiratory viruses in the origin and exacerbations of asthma. AB - PURPOSE OF REVIEW: The present review focuses and comments on the increasing body of evidence correlating respiratory viral infections with asthma onset and exacerbations. RECENT FINDINGS: Recent data suggest multiple and some time contrasting roles for viral infection in the origin of asthma. These data also indicate that the immune status of the host, including atopy, may interactively contribute to this process, conferring susceptibility or even resistance to the development of asthma in virus-infected individuals. In the presence of asthma, the role of viral infection in triggering exacerbations is clearly established. Chemokine and cytokine responses of the respiratory epithelium, a biased type 1/type 2 cytokine balance, defective costimulation, as well as abnormal neural control have been suggested as possible mechanisms. The importance of concurrent or synergistic effects of allergen exposure is currently under scrutiny. SUMMARY: Viruses may initiate and certainly exacerbate asthma. Mild repeated infections early in life could also stimulate type 1 immune responses conferring protection from atopy and asthma. The host's immune status, the type of viral infection and the timing of exposure to various environmental stimuli are probably the key factors in this process. Mechanistic insights deduced from recent work should allow for the development of intervening strategies in the near future. PMID- 12582314 TI - The role of the mast cell in asthma: a reassessment. AB - PURPOSE OF REVIEW: Interest in the role of the mast cell in bronchial asthma has waxed and waned over several decades, but there is now compelling evidence that mast cells make an important contribution to the pathophysiology of this disease. This review will discuss current advances in this field. RECENT FINDINGS: Mast cells, but not T cells or eosinophils, localize within the bronchial smooth muscle bundles in patients with asthma but not in normal individuals or those with eosinophilic bronchitis. Smooth muscle mast cell density correlates significantly with indices of bronchial hyperresponsiveness, and is likely to be an important factor determining the asthmatic phenotype. Tryptase induces proliferation in human airway smooth muscle, and the recently identified transmembrane form induces the development of bronchial hyperresponsiveness in mice. IL-4 and IL-13, known mast cell products, also induce bronchial hyperresponsiveness in the mouse, in the absence of an inflammatory response. There are therefore several pathways by which the close approximation of mast cells with airway smooth muscle cells might lead to disordered smooth muscle function. Mast cells also infiltrate the airway mucous glands in patients with asthma, showing features of degranulation, and a positive correlate with the amount of mucus obstructing the airway lumen. Taken together these observations suggest that mast cells also play an important role in regulating mucous gland secretion. SUMMARY: The development of potent and specific inhibitors of mast cell secretion, which remain active when administered long term to asthmatic airways, should offer a novel approach to the treatment of asthma. PMID- 12582315 TI - The relationships between atopy, rhinitis and asthma: pathophysiological considerations. AB - PURPOSE OF REVIEW: A close relationship has been described between atopy, allergic rhinitis and asthma. The purpose of this work was to review recent data that have become available on the interactions between these conditions and the ways in which they influence one another. RECENT FINDINGS: Recent findings support previous observations suggesting that atopic dermatitis and rhinitis often accompany or precede the development of asthma. Further data support the notion that early-life exposure to domestic animals, a farming environment, passive smoking, and being raised in a large family, may be protective against the development of atopy and/or allergic diseases, although this seems modulated by genetic factors. Furthermore, the appearance of house-dust-mite-specific immunoglobulin E antibodies in early childhood has been identified as a major risk factor for the development of asthma in children with atopic dermatitis; and the association between sensitization to specific allergens and airway hyperresponsiveness was reported to be the strongest for indoor allergens such as house-dust-mite and cat. Allergen exposure can increase airway responsiveness in non-asthmatic subjects with allergic rhinitis and is associated with an increase in markers of lower airway inflammation, particularly with indoor allergens. Furthermore, nasal allergen provocation can induce bronchial inflammation and vice versa, suggesting close interrelations between upper and lower airways. SUMMARY: In summary, the recent observations on the relationships between atopy, rhinitis and asthma support the hypothesis of a unique systemic condition with variable manifestations, which may develop following an imbalance between T helper cell types 1 and 2 lymphocyte populations. The latter may be influenced by environmental exposure in early life. Upper- and lower-airway inflammatory events influence each other, supporting the concept of 'united airways'. Further studies should look at the relationships between these conditions to identify individuals at high-risk of developing them and develop strategies to possibly prevent their onset. PMID- 12582316 TI - Extending the understanding of leukotrienes in asthma. AB - PURPOSE OF REVIEW: Leukotriene modifiers have recognized utility in the management of asthma. The aim of this review is to put into context recent research findings that extend our understanding of cysteinyl leukotriene synthesis and actions in the pathogenesis of asthma and allergic disease. RECENT FINDINGS: Previous literature has shown that T helper type 2 cytokines thought to favor asthma and allergic diseases upregulate leukotriene synthesis. Recent findings show that interleukins-4 and -13 also upregulate cysteinyl leukotriene 1 receptor expression. Conversely, the regulation of cytokine expression by leukotrienes has also been explored: cysteinyl leukotrienes upregulate type 2 cytokine expression and decrease type 1 cytokine expression, favoring an allergic phenotype. Genetic determinants of the expression of leukotriene-forming enzymes include polymorphisms of the 5-lipoxygenase and LTC(4) synthase promoters. Novel actions of leukotrienes continue to be recognized, and a role for leukotrienes in the development of airway remodeling accompanying chronic asthma is discussed. Mounting evidence implicates leukotrienes in the pathogenesis of asthma following viral infections. Finally, advances in the measurement of leukotrienes are reviewed. SUMMARY: Leukotrienes and their receptors play an important role in the pathogenesis of asthma. Advances in our understanding of the synthesis and actions of these lipid mediators provide the scientific rationale for appropriate utilization of leukotriene modifiers and for envisioning novel leukotriene-based therapeutic approaches in the clinical management of asthma. PMID- 12582317 TI - The role of adenosine as a novel bronchoprovocant in asthma. AB - PURPOSE OF REVIEW: There is evidence that bronchial hyperresponsiveness to inhaled adenosine may be a better marker of airway allergic inflammation than bronchial hyperresponsiveness to other spasmogens. Here the authors review the most recent development in this area of rapidly evolving clinical research, focusing on the putative role of adenosine as a useful bronchoprovocant in asthma. RECENT FINDINGS: New studies have provided evidence that hyperresponsiveness to AMP strongly correlates with sputum, blood/bronchial tissue eosinophilia and exhaled nitric oxide, whereas hyperresponsiveness to methacholine is related to changes in forced expiratory volume in 1 s, thus reflecting changes in airway caliber and functional state. Other recent studies have emphasized that the state of atopy is critically important in the bronchial response to inhaled AMP. The early finding that in people with asthma treatment with inhaled glucocorticosteroids significantly reduces airway responsiveness to AMP, as compared with other spasmogens such as methacholine or bradykinin, has been confirmed by recent reports which have also outlined that this is an early phenomenon. SUMMARY: Taken together these studies evidence that hyperresponsiveness to inhaled adenosine may be a better marker of airway allergic inflammation than hyperresponsiveness to other spasmogens. This peculiar property of adenosine suggests that bronchial provocation with inhaled adenosine could provide a reliable noninvasive tool for monitoring asthma activity/progression, and assessing the response to antiinflammatory treatments. PMID- 12582318 TI - Genetics and epidemiology. PMID- 12582319 TI - The value of isolated populations in genetic studies of allergic diseases. AB - Asthma and other IgE-mediated atopic disorders are extremely common in the population, with a consistent, but rather small, genetic component (lambda 3-5) in the inheritance of the disease. So far, genome-wide gene mapping studies in asthma-related traits have not achieved the same success as the positional cloning of genes for Mendelian diseases. Many disease predisposing alleles are believed to be involved in the development of these phenotypes, but at present we do not understand their genetic architecture. As a result of reduced allelic diversity and longer-range linkage disequilibrium compared with more mixed populations, isolated populations may offer a unique opportunity to model a simplified picture of these complex genetic events predisposing to common diseases, such as asthma, which demonstrate familial clustering, but do not follow a simple Mendelian mode of inheritance.(sib) PMID- 12582320 TI - The genetics of atopic dermatitis. AB - PURPOSE OF REVIEW: Atopic dermatitis is typified by itchy, inflamed skin. It is increasingly common in the developed world and is a major cause of morbidity in infants and young children. Most children with the disease have high levels of immunoglobulin E and many have concomitant asthma. The cause of the disease is unknown, but it is highly heritable. Identification of the genes and genetic variants underlying atopic dermatitis may lead to new treatments and better classification of children with the disease. RECENT FINDINGS: Preliminary genetic studies have identified genes or clusters of genes that are expressed in the outermost layer of the skin to be just as important as genes that may modify the atopic process. These genes may influence other diseases, including psoriasis. Genome screens in mouse models seem to indicate involvement of some of the equivalent chromosomal regions as for human disease. SUMMARY: The findings suggest that atopy in atopic dermatitis may be a secondary process, rather than the cause of the disease. The barrier function of the skin is seen not to be merely passive. Identification of the genes underlying atopic dermatitis is feasible and likely within a few years. PMID- 12582321 TI - Genetics of IL-13 and functional relevance of IL-13 variants. AB - Studies in both humans and mice clearly show that interleukin (IL)-13 is a central regulator of allergic inflammation. Numerous single nucleotide polymorphisms have been recently identified in the IL-13 gene and have been found to be associated with allergic and/or asthmatic phenotypes in different populations throughout the world. Furthermore, functional genomics studies are highlighting mechanistic pathways that may link genetic variation in IL-13 and disease. IL-13 is therefore an ideal model to study the impact of genetic variation on the regulation of gene expression and function in complex diseases. PMID- 12582322 TI - Pharmacogenetics of treatment with leukotriene modifiers. AB - PURPOSE OF REVIEW Pharmacogenetics is emerging as a field with great potential to improve both our understanding and treatment options in asthma. This review highlights the importance of pharmacogenetic associations of an important class of asthma therapy, the leukotriene modifiers, and asthma. RECENT FINDINGS: Over the past decade, different leukotriene modifier therapies have emerged and have resulted in improvements in asthma parameters in individuals with this condition. However, there is substantial interindividual variability with respect to the response to this and other asthma therapies. Over the past few years, polymorphisms of two genes in the leukotriene pathway, the gene and the synthase gene, have been identified and have been demonstrated to have pharmacogenetic associations with asthma. However, currently identified genetic determinants explain the response to therapy in only a minority of patients. SUMMARY: As the field of pharmacogenetics advances, an increasing proportion of individual variation in response to pharmacotherapy will be predictable on the basis of associations with particular genetic polymorphisms or patterns of polymorphisms. The pharmacogenetic association of leukotriene modifiers and asthma is an excellent example of how these associations hold out the promise of being able to individualize pharmacotherapy, by providing specific medications to those most likely to respond while avoiding therapy in those most likely to suffer adverse effects. PMID- 12582323 TI - New diagnostic classification of allergic skin disorders. AB - PURPOSE OF REVIEW: To highlight the importance of one common nomenclature for allergic skin disorders. RECENT FINDINGS: Diseases are best defined on the pathophysiological mechanisms underlying the symptoms. Although the mechanisms behind what is sometimes unfortunately referred to as 'atopic eczema' or 'atopic dermatitis' have been quite well understood for 30 years the present terms used are confusing. Over the years more than 20 different names have been used. SUMMARY: Doctors, scientists and patients in the field of allergy must use one and the same terminology for allergic diseases or the result will be confusion. Diagnosis, prevention, treatment, patient advice, environmental relations, and the search for genetic markers are examples of areas that will be hampered by conflicting results. Strong efforts should be invested into broadly implementing the European Academy of Allergology and Clinical Immunology (EAACI) revised nomenclature for allergy. PMID- 12582325 TI - Conjunctival provocation testing: overview of recent clinical trials in ocular allergy. AB - PURPOSE OF REVIEW: The conjunctival provocation test is a human model of ocular allergy that has been used to study the ocular response to allergenic stimuli and to evaluate antiallergic therapy. The review will discuss recent studies using the conjunctival provocation test and other models of ocular allergy, and evaluate the relative merits of different models. RECENT FINDINGS: The conjunctival provocation test has allowed investigators to recruit large numbers of allergic individuals who, although asymptomatic, can be challenged with the conjunctival administration of allergen. Observations of the eye can be made before and after challenge, and cells and mediators may be sampled from the ocular surface. In addition, the effectiveness of antiallergic therapy can be evaluated, usually by pretreating the two eyes with different forms or doses of drug. SUMMARY: Considerable useful information has been gained about the ocular allergic response and drug efficacy using the conjunctival provocation test and naturally occurring seasonal allergic conjunctivitis. PMID- 12582324 TI - Recent developments in latex allergy. AB - PURPOSE OF REVIEW: Immediate allergic reactions to natural rubber latex continue to be an important medical and occupational health problem. In this review we focus on progress made in understanding the significance of occupational exposure and epidemiology, risk groups, diagnosis and prevention of natural rubber latex allergy. We also discuss methods aimed at quantification of clinically relevant natural rubber latex allergens and studies on B-cell epitopes of major natural rubber latex allergens. RECENT FINDINGS: Prospective studies have been published focusing on the prevention of natural rubber latex allergy. Efforts to identify and characterize new natural rubber latex allergens were continued in several research groups. The use of purified allergens or proteins produced by recombinant DNA technology was assessed in studies aimed at improving the diagnosis of natural rubber latex allergy as well as in developing quantitative methods for the measurement of specific natural rubber latex allergens. For the first time, conformational immunoglobulin E epitopes were identified in a major natural rubber latex allergen, hevein, using a novel chimera-based allergen epitope mapping strategy. SUMMARY: Measures taken in health care to reduce exposure to natural rubber latex products seem to be effective in reducing the number of new sensitizations. A few new minor natural rubber latex allergens, probably important in allergen cross reactions, were identified, and some nonmedical natural rubber latex products were found to be potentially dangerous to natural rubber latex-allergic patients. Sensitive and specific immunoassays for quantification of clinically relevant natural rubber latex allergens in manufactured products were developed in some laboratories. These assays may eventually replace the nonspecific total protein measurement, currently advocated by health authorities in the indirect estimation of allergen amounts in natural rubber latex products. Knowledge about conformational immunoglobulin-binding B cell epitopes of major allergens is expected to be helpful in designing optimal reagents to specific immunotherapy. PMID- 12582326 TI - Human ocular mast cells. AB - PURPOSE OF REVIEW: Seasonal allergic conjunctivitis is a common and clinically significant type I hypersensitivity response, in which the mast cell is considered to play a pivotal role in causing the signs and symptoms, including ocular itching, hyperaemia, lacrimation and chemosis. This review focuses on the biology of the human mast cell, particularly that of the human conjunctiva. RECENT FINDINGS: The ocular mast cell not only releases histamine and eicosanoids into the extravascular environment when activated, but also synthesizes the cytokines IL-4 and TNF-alpha. The number of IL-4 messenger RNA-positive mast cells found in the conjunctival submucosa increases threefold in seasonal allergic conjunctivitis 'in season' compared with 'out of season', suggesting a role in disease. Treatment of the eye with 2% nedocromil sodium eye drops twice a day for 2 weeks reduced the tear concentrations of both histamine and prostaglandin D(2) by more than 70% at 30 min after challenge (both <0.05) illustrating an effective mast cell stabilizing effect in the conjunctiva. SUMMARY: Mast cells are a heterogeneous family of cells that are pivotal in initiating the signs and symptoms of seasonal allergic conjunctivitis. The expression of cytokines also endows them with the ability to initiate the inflammatory cascade, resulting in eosinophil accumulation associated with vernal conjunctivitis. Drug modulation of mast cell activity, although reducing the acute symptoms of active disease, also reduces the cytokine stimulus for the development of chronic allergic inflammation. PMID- 12582327 TI - Spectrum of ocular allergy. AB - PURPOSE OF REVIEW: To elaborate and review the immunologic spectrum of the five basic types of ocular allergy. RECENT FINDINGS: Perennial allergic conjunctivitis (PAC) appears to be an extension of seasonal allergic conjunctivitis (SAC) based on the similarity of clinical symptoms and the immune mechanism involved. T helper type 2 (TH2)-type cytokines, increased ratio of TH1/TH2 cytokines and increased statement of adhesion molecules all appear to play an integral role in the inflammatory process of SAC and PAC. Vernal keratoconjunctivitis (VKC), which is a pediatric disease, differs from SAC and PAC by its more severe and chronic nature. Increased numbers of T cells, eosinophils (along with the products of degranulation), chemokines and their receptors may contribute to the more serious symptoms of VKC. Atopic keratoconjunctivitis (AKC) is similar to VKC by the increased concentration of cytokines involved (i.e. interleukin-4 and -5). Despite clinical similarities with VKC, the presence of atopic dermatitis, along with some differences in clinical signs, make AKC a separate entity, which is sometimes referred to as an adult variant of VKC. Giant papillary conjunctivitis has similar cell involvement as VKC and AKC. However, giant papillary conjunctivitis differs from VKC and AKC by the clinical signs present (i.e. the presence of giant papillae) and the necessary concurrent contact lens wear. SUMMARY: The important differences and similarities observed in these five types of allergic diseases might help to better treat the patients affected with these disorders. PMID- 12582328 TI - Immunomodulatory therapy for anterior segment ocular inflammation. AB - PURPOSE OF REVIEW: To outline immunomodulatory therapy for ocular inflammatory diseases of the anterior segment and to review the current literature regarding their efficacy and safety. RECENT FINDINGS: Most current studies concern immunomodulatory treatment of patients who have chronic, debilitating disease resistant to corticosteroids. The authors evaluated the efficacy of treatment, the side effect profiles and the potential for long-term remission. SUMMARY: Immunomodulatory drugs are increasingly being used in ocular inflammatory disease to avoid the effects of chronic steroid therapy and to induce remission in patients with chronic disease. These therapies have shown efficacy in treating otherwise resistant disease with relative safety insofar as side effects are generally reversible and medications can be substituted if intolerable side effects arise. Future trends in treatment will emphasize more specific target molecules, less immunosuppressive drugs and modification of the immune environment to increase the rates of long-term remission. PMID- 12582330 TI - Current opinion of immunotherapy for ocular allergy. AB - PURPOSE OF REVIEW: Allergic conjunctivitis is common and may be the most prominent or the only feature of allergies. Immunotherapy has been used as a primary treatment for allergies since the early 1900s. Currently the use of immunotherapy for allergic rhinoconjunctivitis is well established and has been shown to decrease the development of bronchial hyperreactivity and asthma. However, the role of immunotherapy for primary treatment of allergic conjunctivitis is unclear. We reviewed the studies where immunotherapy was used with particular attention to the affects on ocular allergies. RECENT FINDINGS: There are many schedules and methods of delivering immunotherapy. Recent studies have started to assess ocular symptoms as one of the parameters to monitor efficacy of therapy. They follow the affects of immunotherapy on conjunctival provocation tests, ocular symptoms, or the use of eye drops. The literature suggests that using the various immunotherapy modalities at different schedules, ocular symptoms improved even when immunotherapy was used on a rush schedule. SUMMARY: The initiation of immunotherapy for allergic rhinoconjunctivitis has been shown to switch the immune response to T helper 1 and thus avoid the progression of other atopic conditions. Current literature shows that using many allergens with different forms of immunotherapy appear to have a significant improvement in ocular allergy symptoms and this can be achieved rapidly and safely in most patients. Whether using immunotherapy early in allergic conjunctivitis will alter the progression of other atopic conditions remains to be investigated. PMID- 12582329 TI - Topical immunopharmacology of ocular allergies. AB - PURPOSE OF REVIEW: To review the histamine-1 receptor antagonists, mast cell blockers and natural agents with such actions that can be used for the topical treatment of ocular allergies. RECENT FINDINGS: Increasing evidence indicates that some histamine-1 receptor antagonists have additional actions to inhibit secretion of inflammatory mediators, especially cytokines, from ocular mast cells and other cell types. Emerging information suggests that such actions may be through regulation of intracellular calcium ion levels of NF-kappaB activation. SUMMARY: A number of available drugs and natural non-prescription agents may have anti-histaminic and anti-inflammatory actions. PMID- 12582332 TI - Nursing humanities: the time has come. PMID- 12582333 TI - Family planning options. PMID- 12582334 TI - From overtime to understaffing. PMID- 12582335 TI - Reflections. Role reversal. PMID- 12582336 TI - Myths and facts about back injuries in nursing. PMID- 12582337 TI - ASC-US and HPV testing. PMID- 12582338 TI - Emergency: gang members in the ED. PMID- 12582339 TI - Phlebitis. PMID- 12582340 TI - Support for parents during a child's hospitalization. PMID- 12582342 TI - Dofetilide and atrial fibrillation. PMID- 12582343 TI - Protecting patient information. PMID- 12582345 TI - Childhood vaccines: how safe are they? PMID- 12582344 TI - Tramadol. PMID- 12582346 TI - Acetaminophen's hidden dangers. PMID- 12582347 TI - School nursing: a successful experiment. PMID- 12582348 TI - The needlestick law. PMID- 12582349 TI - The great teaching case. PMID- 12582350 TI - Anemia. PMID- 12582351 TI - Cheilitis glandularis: An unusual presentation in a patient with HIV infection. AB - Cheilitis glandularis is a rare disorder of unknown etiology characterized by inflammation of the minor salivary glands of the lower lip. The present report details the features of a patient who presented with cheilitis glandularis and was subsequently found to also have undiagnosed HIV infection. PMID- 12582352 TI - Prophylactic single-dose administration of 600 mg clindamycin versus 4-time administration of 600 mg clindamycin in orthognathic surgery: A prospective randomized study in bilateral mandibular sagittal ramus osteotomies. AB - OBJECTIVE: The purpose of this study was to compare a single 600-mg dose of preoperative intravenously administered clindamycin with a 24-hour 600-mg regimen of clindamycin as prophylaxis for postoperative infections in bilateral sagittal ramus osteotomies. STUDY DESIGN: Seventy patients were assigned at random to an antibiotic protocol. Postoperative infection was assessed by one clinician blinded to the protocol. All data were tabulated, and a statistical analysis was performed by means of the paired t test. RESULTS: In 3 patients, an infection developed. Two of those patients belonged to the single-dose clindamycin group and the other to the 24-hour clindamycin prophylaxis group. The sutures were removed, and wound exudate specimens were obtained for Gram staining and culture. After a 5-day period of 500 mg amoxicillin 4 times daily, the parameters of infection normalized. CONCLUSIONS: There were no statistically significant differences between postoperative infection after the single dose of clindamycin and infection in the 24-hour prophylaxis group (P =.3244). PMID- 12582353 TI - Comparison of conventional surgery with motorized trephine in bone harvest from the anterior iliac crest. AB - OBJECTIVE: A prospective study was performed to compare morbidity associated with the harvest of corticocancellous block grafts (CCBG) by conventional surgery and procurement of cancellous cores (CC) by means of a motorized trephine from the anterior ilium. STUDY DESIGN: Seventy-six patients requiring 30 mL or less of cancellous bone for maxillofacial reconstruction were placed into 2 treatment groups. One group underwent harvest of CCBGs in the traditional open medial approach to the anterior ilium. The second group had CCs harvested through a 0.5- to 1.0-cm incision with a motor-driven trephine. The following parameters were used to evaluate patient morbidity: number of days to unassisted ambulation, length of hospital stay, and pain scores for both the recipient and the donor sites. RESULTS: The mean time to patients' unassisted ambulation following a CCBG was significantly longer (2.8 days) than following CC (0.8 days). The mean length of hospital stay following a CCBG was significantly longer (4.1 days) than following a CC (2.2 days). The mean contemporaneous maxillofacial pain scores following procurement of CCs (day 1: 5.6; day 3: 4.2) and procurement of CCBGs (day 1: 5.8; day 3: 4.5) were not significantly different, whereas the mean hip pain was significantly greater (P <.05) on day 1 in patients having undergone a CCBG (pain score: 6.2) than in patients having undergone a CC graft (pain score: 3.0). Gait disturbance was identified in 15 of 22 patients (68%) who underwent a CCBG. Only 1 of the 54 patients (1.9%) who underwent a CC graft exhibited an abnormal gait. No other complications were noted. The results demonstrated a significant difference in morbidity between the 2 techniques in all variables. CONCLUSION: Where modest amounts of cancellous bone are required for maxillofacial grafting, trephining of cancellous cores results in significantly less morbidity than traditional open methods. PMID- 12582354 TI - A new surgical approach to extensive tumors in the pterygomaxillary fossa and the skull base. AB - OBJECTIVE: The article describes a new surgical approach to extensive tumors in the pterygomaxillary fossa and the middle skull base. METHODS: The first incision divides the upper lip in the midline, passes under the nasal pyramid, and extends laterally, reaching the level of the temporamandibular joint, at which point it exits to meet the vertical coronal/preauricular incision. An incision is then made along the maxillary buccogingival fold on the involved side, running from the midline to the retromolar area. Another incision is made along the mandibular buccogingival fold on the involved side, running from canine to retromolar area. The cheek flap is reflected inferiorly to the level of the angle of mandible after the elevation of the maxillary and mandibular periosteum and the messeteric fascia in a downward fashion. The frontotemporal scalp is reflected toward the midline after completion of the coronal and transtemporal incision and an appropriate undermining. The craniofacial skeleton is exposed from the midline. After osteotomies of the orbitomaxillary skeleton and a horizontal subperiosteal osteotomy at the angle of the mandible are completed, a frontotemporal craniotomy is performed, and the foramina spinosum, ovale, and rotundum, as well as the superior orbital fissure, are identified. The tumor is exposed in the surgical field; after the tumor is resected, the orbitomaxillary skeleton and the mandibular namus, removed during the approach phase, are replaced and affixed by miniplate. The external soft tissues, including the skin, are then reapproximated along the preoperative markings. RESULTS: The surgical field obtained at the skull base extends from the contralateral eustachian tube to ipsilateral geniculate ganglion. It includes the nasopharynx, clivus, sphenoid, cavernous sinus, and the entire infratemporal fossa, as well as superior orbital fissure and the pterygopalatine fossa. This approach was used in 5 patients. All patients healed per primum. CONCLUSION: This technique is especially useful for excising tumors in the nasopharynx, clivus, superior orbital fissure-cavernous sinus, and infratemporal and pterygopalatine fossa. PMID- 12582355 TI - Smoking behavior and views of Jordanian dentists: A pilot survey. AB - OBJECTIVE: The purpose of this study was to examine tobacco use among Jordanian dentists and their views on smoking. STUDY DESIGN: In July 1999, a self-reported questionnaire survey was mailed to a representative sample of 849 of 1693 licensed general dentists. The data were analyzed by using the Statistical Package for the Social Sciences (Chicago, Ill), and the statistical significance was determined by means of the chi(2) test. RESULTS: A response rate of 72.2% (613) was obtained. Overall, two thirds of the respondents were men younger than 40 years of age who worked in the private sector. Current tobacco users made up 35% (215/613), the majority of whom smoked cigarettes, with 83% being daily smokers. One fifth of daily smokers said that they smoke 20 or more cigarettes per day. Of respondents, 86.8% agreed that dentists should be a nonsmoking role model, and 77% believed that they should be involved in tobacco cessation counseling. Only 38.3% thought that they could convince patients who smoke to quit. Nonsmokers were significantly more likely to respond positively to these questions. Most respondents (92.2%) reported that the harmful effect of tobacco on health is a good reason to motivate smokers to quit. Unfortunately, almost half of the respondents (46.7%) thought that dental treatment is more important than providing tobacco counseling, and those were significantly more likely to be from private practices. CONCLUSIONS: Despite the high proportion of general dentists who were current smokers, many hold positive views on the tobacco issue. An education program targeted at dentists in Jordan is needed to discourage them from using tobacco and to teach them tobacco-cessation techniques and behaviors to reduce use among their patients. PMID- 12582356 TI - Linear IgA dermatosis induced by a new angiotensin-converting enzyme inhibitor. AB - A 68-year-old female patient treated with benazepril for arterial hypertension developed oral and cutaneous blistering. Biopsy of the oral and cutaneous lesions showed neutrophilic microabscesses in the mesenchymal papillae, with epitheliomesenchymal separation. Direct immunofluorescence revealed linear immunoglobulin deposits at the epithelial basement membrane zone, consisting predominantly of IgA. The histologic results supported the clinical diagnosis of drug-induced linear IgA disease. The substitution of benazepril with a beta blocker resulted in complete resolution of all mucocutaneous lesions. PMID- 12582357 TI - Specific expression of inducible nitric oxide synthase in the synovium of the diseased temporomandibular joint. AB - OBJECTIVE: The expression of inducible nitric oxide synthase (iNOS) in temporomandibular joint (TMJ) specimens obtained arthroscopically from diseased TMJs was investigated by using immunohistochemistry and compared with clinical, arthroscopic, and histologic findings. STUDY DESIGN: Synovial biopsies were obtained arthroscopically from 18 TMJs in 15 patients with symptomatic internal derangement (ID) or osteoarthritis (OA). We also obtained arthroscopic biopsies from 8 control TMJs (3 with habitual luxation of the mandible, one with ID with clicking, and 4 with mandibular condyle fractures). The expression of iNOS was examined by immunohistochemistry and was compared with clinical, arthroscopic, and histologic findings. RESULTS: Definite or intense iNOS immunoreactivity was observed in both the synovial lining cells and the endothelial cells of TMJs with symptomatic ID or OA. Weaker immunoreactivity was present in synovial fibroblasts. In contrast, in TMJs without synovitis (eg, those with habitual luxation of the mandible) the expression of iNOS was weak or marginal. The expression of iNOS correlated significantly with arthroscopic evidence of synovitis (r = 0.406, P <.05) but not with cartilaginous degeneration (P >.05). The expression of iNOS also correlated with the histologic grade of the synovial lining cell layers (r = 0.530, P <.05). However, in patients with ID or OA of the TMJ, there was no statistically significant correlation between the intensity of iNOS immunoreactivity and clinical, arthroscopic, or histologic findings (P >.05). CONCLUSION: These data clearly suggest that nitric oxide is locally produced in the synovial lining of the TMJ in ID and OA. PMID- 12582358 TI - Modification to the approach of the diagnosis of mucous membrane pemphigoid: A case report and literature review. AB - Mucous membrane pemphigoid (MMP) is a subepithelial blistering disease predominantly involving the mucosal surfaces. Distinct subgroups of MMP have recently been identified by using advanced immunopathologic and immunochemical techniques and are described in the literature on the basis of their clinical features and antigenic specificities. Antiepiligrin MMP has been described as an immunochemically distinct entity. Evidence of an existing relationship between malignancy and antiepiligrin MMP has been well documented in the medical literature. This case report illustrates a case of antiepiligrin MMP in a patient with an advanced cancer of the bladder. PMID- 12582359 TI - Nucleolar organizer regions in selected odontogenic lesions. AB - OBJECTIVE: The present study aimed to assess the usefulness of argyrophilic nucleolar organizer regions (AgNORs) as quantitative criteria in the diagnosis of odontogenic cysts and tumors. STUDY DESIGN: This descriptive cross-sectional study was done on archival paraffin blocks (n = 37), consisting of 10 conventional ameloblastomas, 7 cases of unicystic ameloblastoma, 10 odontogenic keratocysts, and 10 dentigerous cysts, all accessioned between 1994 and 2000. RESULTS: The findings showed a significant statistical difference among the 4 lesions (P <.001). Conventional and unicystic ameloblastomas had a significantly higher (P <.05) number of AgNORs than odontogenic keratocysts and dentigerous cysts. No significant differences were observed within each group of tumors or cysts. The number of AgNORs was also significantly higher (P <.005) in the nuclei of basal layer cells of the epithelial lining of keratocysts than in the nuclei of parabasal layer cells. AgNORs in conventional and unicystic ameloblastomas were smaller but more broadly distributed, which may indicate higher proliferative activity. CONCLUSION: AgNORs can be useful in the histopathologic differentiation of ameloblastomas from odontogenic cysts. PMID- 12582360 TI - Recurrent multiple superficial mucocele on the palate: Histopathology and laser vaporization. AB - A 55-year-old Japanese woman presented with recurrent multiple vesicles on the soft palate. Some vesicles ruptured spontaneously but soon recurred, and the patient felt mild pain and discomfort. Histopathologically, intraepithelial mucocele with mucus retention in the salivary ducts was observed. The lesion was successfully treated with Nd: YAG Contact Laser. There has been no recurrence for 3 years. PMID- 12582361 TI - Peripheral clear cell variant of calcifying epithelial odontogenic tumor: Report of a case and immunohistochemical investigation. AB - A case of peripheral calcifying epithelial odontogenic tumor, clear cell variant, located on the right gingival maxilla of a 48-year-old woman, presenting as a 2.0 cm solitary, firm nodule was studied. Microscopically, it was composed of polyhedral and clear epithelial cells associated with amyloid-like deposition. The clear epithelial cells exhibited granules that were positive for periodic acid-Schiff, and the amyloid-like deposit stained with Congo red showed a green birefringence in the polarized light. Polyhedral and clear epithelial cells were immunopositive for AE1/AE3 and cytokeratin 14. Immunoexpression of fibronectin and types I and III collagen were different between the amyloid-like deposits and the connective tissue stroma. Tenascin surrounded epithelial cells located inside the amyloid-like deposits. Laminin and type IV collagen were immunodetectable around the strands, cords, and nests of epithelial cells. This report represents the seventh case of peripheral calcifying epithelial odontogenic tumor, clear cell variant. PMID- 12582362 TI - Trends in the incidence of oral cancer in Nova Scotia from 1983 to 1997. AB - OBJECTIVE: The purpose of the study was to characterize the total burden of oral cancer in Nova Scotia over the 15-year period from 1983 to 1997 and to identify any trends in the number of cases or incidence rates at specific anatomic sites or within specific age or sex groups over this time period. STUDY DESIGN: All cases that had a diagnosis of invasive oral cancer (ICD-9 sites 140-146) during the study period were retrieved from the records of the Nova Scotia Cancer Registry, which theoretically includes all cancer cases in the province. Cases of in situ carcinoma, lymphoma, and leukemia were not included. All cases during the study period were categorized by tumor site and the age and sex of the patient. Tumors at ICD-9 sites 141 and 143-146 were analyzed separately as intraoral cancer. Trends were studied by grouping cases into three 5-year periods: period 1 (1983-1987), period 2 (1988-1992), and period 3 (1993-1997). Age-standardized incidence rates were calculated to the 1991 Canadian standard population. RESULTS: A total of 1,155 cases of oral cancer were registered in Nova Scotia for this 15-year study period, accounting for 2.0% of all cancer cases. There was an average of 57 cases in men and 20 cases in women per year. Overall, the most common site was the lip (26% of cases), followed by the tongue (20%), other mouth (16%), tonsil/oropharynx (12%), salivary gland (12%), floor of mouth (10%), and gum (4%). About 5% of cases occurred in patients 40 years and younger, and 53% occurred in patients 65 years and older. Lip cancer in men decreased by 38%, and the age-standardized incidence rate fell from 5.6 to 3.0 per 100,000 from period 1 to period 3. Intraoral cancers in men increased by 23% and the age-standardized incidence rate increased by 10% from period 1 to period 3. Both measures peaked in period 2. Intraoral cancer in women increased steadily by 84%, and the age standardized incidence rate increased by 48% from period 1 to 3. The male-to female ratio for intraoral cancer cases decreased from 2.9 to 2.5 to 1.9 over the 3 periods. CONCLUSION: Although there was a decreasing trend for lip cancer, the number of cases and the age-standardized incidence rate for intraoral cancer increased over the 15-year study period. Intraoral cancer increases in females were dramatic. Trends in the number of cases reflected changing risk and a growing, aging population. More oral cancers occurred each year than cancers of the uterine cervix, suggesting the need for more resource allocation in the areas of research, prevention, and early detection of oral cancer. PMID- 12582363 TI - Proliferating cell nuclear antigen and Ki67 immunoreactivity in ameloblastomas. AB - OBJECTIVE: This study used proliferating cell nuclear antigen (PCNA) and Ki67 to evaluate and compare the in situ proliferative activity of (1) solid and multicystic and (2) unicystic ameloblastomas in an attempt to provide a scientific basis for any differences in biologic behavior that exists between these 2 groups of lesions. STUDY DESIGN: Twenty archival tissue sections, 10 of primary unicystic and 10 of solid and multicystic ameloblastomas, were immunohistochemically stained with PCNA and Ki67 antisera. Immunoreactivity was evaluated by the counting of cells, and the data obtained were statistically analyzed. RESULTS: The results showed that cellular proliferative activity varied within the ameloblastoma types. The unicystic ameloblastomas showed statistically significantly higher PCNA and Ki67 labeling indices than the solid and multicystic variant. CONCLUSION: There appears, therefore, to be no correlation between proliferative activity as shown by these proteins and reported biologic behavior. PMID- 12582364 TI - Attachment of human periodontal ligament fibroblasts to 3 different root-end filling materials: Scanning electron microscope observation. AB - OBJECTIVE: The attachment behavior of the human periodontal ligament (PDL) fibroblasts to root-end filling materials (amalgam and Super-EBA) was compared in vitro to gutta-percha by means of scanning electron microscope. STUDY DESIGN: Amalgam and Super-EBA were placed in a prepared cavity of root slices of freshly extracted human teeth and evaluated freshly prepared. Root slices of teeth with cold-burnished gutta-percha filling with AH26 sealer were used for comparison. The root slices were placed in tissue culture cluster, and 1 mL of cell suspension was added carefully over the root slice. They were incubated at 37 degrees C and 100% humidity for 4, 24, and 72 hours. RESULTS: Results showed that the cold-burnished gutta-percha provides a better substrate than amalgam and Super-EBA for cell growth and attachment. Amalgam was the most toxic material, showing early manifestation of cell injury. CONCLUSION: It was concluded that the composition and surface texture of the substrate have an influence on the morphology and the attachment of the PDL fibroblasts. It is suggested that cell attachment and morphology might reflect the biocompatibility of the substratum. PMID- 12582365 TI - A comparison of the shaping abilities of 4 nickel-titanium rotary instruments in simulated root canals. AB - OBJECTIVE: The purpose of this study was to compare the root canal shaping abilities of 4 nickel-titanium rotary instruments. STUDY DESIGN: ProFile, GT Rotary, Quantec, and ProTaper files were used to instrument 48 simulated curved root canals in plastic blocks with the crown-down technique. One operator prepared all the canals until reaching an apical canal size of #30. The instrumentation time, changes of canal dimension and curvature, canal aberration, and instrument deformation were evaluated. Data were analyzed by using one-way analysis of variance followed by the Duncan multiple range test. RESULTS: The ProTaper took significantly less instrumentation time, removed more canal wall (especially at the inner side of the canal curve), lessened the canal curvature, and induced more instrument deformation than did the other instruments. CONCLUSION: Although ProTaper files cut more canal wall more quickly than the other instruments in the curved canal, they were also the instruments that most frequently became deformed. PMID- 12582366 TI - Decalcifying effect of EDTA-T, 10% citric acid, and 17% EDTA on root canal dentin. AB - OBJECTIVE: An in vitro study was conducted to determine the efficacy of 17% EDTA plus 1.25% sodium lauryl ether sulfate (EDTA-T), 10% citric acid, and 17% EDTA with respect to Ca(++) extraction. STUDY DESIGN: The root canals of 90 extracted human canines were instrumented by using the step-back technique. The teeth were randomly divided into 9 groups of 10 teeth each according to the solution and time intervals involved (ie, 3, 10, and 15 minutes). The Ca(++) concentration of the solution was determined by using atomic absorption spectroscopy. RESULTS: At 3 minutes, there was no significant difference between Ca(++) extraction by 10% citric acid and 17% EDTA (P >.05). EDTA-T had the least efficacy for Ca(++) extraction (P <.05). At 10 minutes and 15 minutes, there were no significant differences between 10% citric acid and 17% EDTA or between EDTA-T and 17% EDTA (P >.05). However, 10% citric acid was significantly better than EDTA-T in terms of Ca(++) extraction (P <.05). CONCLUSION: Both 10% citric acid and 17% EDTA are good decalcifying agents when the step-back technique is used. PMID- 12582367 TI - Clinical study of a spacer to help prevent osteoradionecrosis resulting from brachytherapy for tongue cancer. AB - OBJECTIVE: We sought to describe a simple method to construct a spacer and to evaluate with the use of computed tomography the spacer's effectiveness in preventing osteoradionecrosis of the mandible. STUDY DESIGN: Fifty-three patients with oral tongue cancers who were treated by means of interstitial brachytherapy were included in this study. Patients underwent a computed tomography examination immediately after the implantation of radioactive sources, with the spacers in place. Distances between the radioactive sources and the lingual surfaces of the mandible were measured on transverse computed tomographs and were evaluated in terms of the development of osteoradionecrosis in the mandible. RESULTS: Statistically significant differences in the frequency of osteoradionecrosis were observed between patients who had received spacers equal to or thicker than 5 mm and those who had received spacers less than 5 mm thick. CONCLUSION: A spacer should have a minimum thickness of 5 mm on its lingual flange to prevent the development of osteoradionecrosis of the mandible. PMID- 12582368 TI - Trifid mandibular condyle: A case report. AB - A few cases of bifid, but no case of trifid mandibular condyle, have been reported in the literature. This article presents the first reported case of trifid mandibular condyle in a living subject with a history of previous trauma to the temporomandibular joint (TMJ). Additionally, the patient's other condyle was bifid. The patient had no complaint related to the functions of TMJ except for minimal weakness following chewing. The etiology and the prognosis of bifid and trifid condyle are discussed briefly. We considered computed tomography essential to rule out early stages of TMJ pathology in similar cases that resemble trifid mandibular condyle on conventional radiographs. PMID- 12582369 TI - Magnetic resonance imaging diagnosis of the temporomandibular joint in patients with orthodontic appliances. AB - OBJECTIVE: The purpose of this study was to investigate the accuracy of magnetic resonance imaging diagnosis of the temporomandibular joint in those with orthodontic appliances by comparing magnetic resonance images of the temporomandibular joints with and without orthodontic appliances. STUDY DESIGN: Magnetic resonance imaging was performed in a total of 20 temporomandibular joints before and after the insertion of 6 kinds of orthodontic appliances (types 1-6), and the magnetic resonance images were compared. RESULTS: With respect to disk position, the diagnostic accuracy was 80%, 75%, 70%, 70%, 65%, and 60% in order from type 1 through type 6. The distribution of stages for the assessment of condylar configurations was 80%, 55%, 40%, 40%, 20%, and 10% in order from type 1 through type 6. No significant changes were found in the condylar head marrow signals. CONCLUSION: Magnetic resonance imaging is performed in orthodontic patients preferably by using ceramic brackets in the front teeth and direct bonding tubes in the molar teeth while removing arch wires. PMID- 12582370 TI - Intravascular lymphomatosis of the skin as a manifestation of recurrent B-cell lymphoma. AB - Intravascular lymphomatosis (IVL) is a rare type of lymphoma with a poor prognosis. Its distinctive clinical and histopathologic features are generated by the proliferation of neoplastic mononuclear cells within blood vessels. We describe a patient with IVL of the skin as a manifestation of a recurrent diffuse large B-cell lymphoma of ureteral origin. Lymphoma cells were located both within the vessels and the parenchyma in an early cutaneous lesion. After recurrence in the skin, lymphoma cells gradually located only in the vascular lumina. This transition suggests that cells localized within the vessels were selected as a consequence of chemotherapy. Immunohistochemical examination revealed that the expression of surface adhesion molecules of lymphoma cells did not significantly change. The results of polymerase chain reaction revealed that the ureteral and cutaneous tumors were identical in clonality. Our findings suggest that conventional diffuse large B-cell lymphoma can change into IVL. PMID- 12582371 TI - Development of an idlike reaction during treatment for acute pulmonary histoplasmosis: a new cutaneous manifestation in histoplasmosis. AB - Id reactions often occur secondary to release of antigens after initiation of antimicrobial therapy. This reaction commonly occurs during treatment of Trichophyton infections. The diagnosis is usually made clinically and is supported in the presence of an infection and/or initiation of therapy. We report an idlike reaction during therapy for pulmonary histoplasmosis. PMID- 12582372 TI - Tinea faciei mimicking cutaneous lupus erythematosus: a histopathologic case report. AB - Tinea faciei is an uncommon dermatophyte infection. Clinically, it may resemble other dermatoses such as cutaneous lupus erythematosus (CLE), polymorphous light eruption, and contact dermatitis. Histopathology is highly variable, demonstrating a spectrum from mild focal spongiosis to a chronic spongiotic psoriasiform dermatitis with a mixed dermal inflammatory infiltrate and fungi in the cornified layer. We report a case of tinea faciei with histopathology that is characteristic of CLE. PMID- 12582373 TI - Cimetidine therapy for epidermodysplasia verruciformis. AB - A 16-year-old boy with an 8-year history of multiple slightly keratotic papules on his face, trunk, and limbs, is described. On the basis of clinical features, histologic examination of 2 lesions and positivity for HPV 5, a diagnosis of epidermodysplasia verruciformis was made. Treatment with oral cimetidine (40 mg/kg per day) induced a marked improvement after 3 months of therapy, with no relapse at a 6-month follow-up. PMID- 12582374 TI - Multicentric reticulohistiocytosis presenting with clinical features of dermatomyositis. AB - Multicentric reticulohistiocytosis (MRH) is a rare disorder with proliferating histiocytes that develop into multinucleated giant cells with "ground-glass" cytoplasm. The disease presents with a rapidly destructive, sometimes permanently debilitating, polyarthritis and a papulonodular eruption, generally of the face and hands. We present 3 cases of MRH in which the initial clinical diagnosis was thought to be dermatomyositis (DM). The cutaneous findings in these cases included an erythematous, predominantly photodistributed macular and papular eruption clinically consistent with DM. However, skin biopsy specimens revealed the diagnosis of MRH. This previously unreported clinical similarity between MRH and DM is significant because the treatment options and future complications vary greatly between the 2 diseases. Thus, it is important to consider the possibility of multicentric reticulohistiocytosis when evaluating a patient with the clinical diagnosis of dermatomyositis. PMID- 12582375 TI - Linear acral pseudolymphomatous angiokeratoma of children (APACHE): further evidence that APACHE is a cutaneous pseudolymphoma. AB - Acral pseudolymphomatous angiokeratoma of children (APACHE) is a recently recognized, rare clinical entity. We report a case of APACHE in a 9-year-old Japanese girl with a dark red, linear plaque on the posterior area of the right lower leg. Histopathologic findings revealed further evidence that APACHE was a cutaneous pseudolymphoma rather than a vascular neoplasm because of the presence of distinct primary and secondary lymphoid follicles, and thick-walled, long blood vessels lined with prominent plump endothelial cells similar to high endothelial venules in the paracortical area of the lymph node. In addition to previous descriptions, the patient presented with a distinct linear lesion. Vacuolar alteration of the basal cell layer and scattered eosinophilic bodies in the epidermis were observed, which suggested a lichenoid tissue reaction. We believe that APACHE might show a distinct linear pattern accompanied by a lichenoid tissue reaction. PMID- 12582376 TI - African tick bite fever: not a spotless rickettsiosis! AB - African tick bite fever is caused by Rickettsia africae, a newly recognized species from South Africa. We report the case of a patient with an unusual site of a tick bite and discuss cutaneous differences from other spotted fevers that may help dermatologists with clinical diagnosis. PMID- 12582377 TI - Hidradenoma papilliferum occurring on the chest of a man. AB - We describe an unusual case of hidradenoma papilliferum on the chest of a man and discuss the rare sites of the tumor. PMID- 12582378 TI - IgA pemphigus occurring in a 1-month-old infant. AB - An 11-month-old girl had recurrent pruritic vesiculopustular lesions involving the trunk, extremities, and scalp since the age of 1-month. Histopathologic examination revealed an intraepidermal bulla produced by acantholysis and neutrophilic spongiosis. Direct immunofluorescence showed IgA deposits in the intercellular space throughout the epidermis. Circulating anti-intercellular IgA autoantibodies were absent. Dapsone therapy was effective. PMID- 12582379 TI - Colocalization of granuloma annulare and mid-dermal elastolysis. AB - Mid-dermal elastolysis is an elastic tissue disorder of unknown causation. Various elastic tissue abnormalities have been described in granuloma annulare. To our knowledge, we report the first case illustrating the clinical and histologic colocalization of granuloma annulare and mid-dermal elastolysis. PMID- 12582380 TI - CD30 (Ki-1)-positive large-cell cutaneous T-cell lymphoma with secondary xanthomatous changes after radiation therapy. AB - A 72-year-old woman with a large mass on her forehead was found to have primary cutaneous CD30-positive large T-cell lymphoma and was treated with radiation therapy. Five weeks after the completion of radiation therapy, a large, yellow plaque developed on the forehead. Histopathologic examination showed numerous xanthoma-like cells with clear and foamy cytoplasm that infiltrated the upper and mid dermis with no residual lymphoma. To our knowledge, this is the first report of xanthomatous change after radiation therapy for primary cutaneous CD30 positive large cell lymphoma. We believe that the xanthomatous change after radiation therapy probably was caused by lysis of radiosensitive lymphoma cells and histiocytic scavenging of lipid-membrane debris. PMID- 12582381 TI - Melanonychia associated with PUVA therapy. AB - We describe a case of melanonychia induced by PUVA. This phenomenon likely is more common than the few reports in the literature indicate. PMID- 12582382 TI - Perforating follicular hybrid cyst of the tarsus. AB - Follicular hybrid cysts are cutaneous tumors that arise from different portions of the hair follicle. We describe a follicular hybrid cyst of the tarsus that contained features of pilomatricoma and steatocystoma and perforated the palpebral conjunctiva. The unusual location and histopathologic features are discussed. PMID- 12582383 TI - Keratoacanthoma developing in sites of previous trauma: a report of two cases and review of the literature. AB - Keratoacanthoma is a common benign squamous neoplasm that only rarely progresses to metastatic carcinoma. The cause of keratoacanthoma is unclear; however, suspected causes include ultraviolet light, genetic factors, immunosuppression, chemical carcinogens, viruses, and trauma. Keratoacanthomas have been reported only rarely to arise in sites of previous trauma. We report 2 cases of keratoacanthomas arising shortly after episodes of cutaneous trauma and provide a review of the literature of similar cases previously reported. It is likely that an interaction exists between a variety of factors in the development of keratoacanthomas; however, it appears that a predilection exists for these lesions to arise in trauma sites. PMID- 12582384 TI - Cutaneous metastatic malignant fibrous histiocytoma. AB - Malignant fibrous histiocytoma is the most common soft tissue sarcoma, but cutaneous metastasis of this tumor is quite rare. A 30-year-old female patient had several brownish 0.5 x 0.5-cm papules on her right ankle. Histopathologic examination revealed spindle-shaped tumor cells with cellular pleomorphism and high mitosis (5 of 10 high-power fields), suggestive of malignant fibrous histiocytoma. She had a history of left subcutaneous breast mass that was diagnosed as malignant fibrous histiocytoma 3 years earlier. We report a case of malignant fibrous histiocytoma with pleomorphism mixed with spindle-shaped cells and multinuclear giant cells, which metastasized to the skin. PMID- 12582385 TI - Hypertrichosis. AB - Hypertrichosis is hair growth that is abnormal for the age, sex, or race of an individual, or for a particular area of the body. Recognized forms of hypertrichosis are reviewed. Hirsutism, which is male-pattern hair growth in a female or child, is not included in this review. Hypertrichosis is categorized as congenital or acquired, and regional or generalized. Methods of managing hypertrichosis are also briefly reviewed PMID- 12582386 TI - Acral lentiginous melanoma mimicking benign disease: the Emory experience. AB - BACKGROUND: Plantar and subungual melanoma exhibits a higher misdiagnosis rate relative to other anatomic sites. Misdiagnosis and delay in diagnosis are statistically associated with poorer patient outcome. Awareness of atypical presentations of acral melanoma may, thus, be important to decrease misdiagnosis rates and improve patient outcome. METHODS: We conducted a retrospective case review of plantar or lower-extremity subungual melanoma performed at Winship Cancer Center, a tertiary care, referral center affiliated with Emory University, between 1985 and 2001. RESULTS: A total of 53 cases of plantar or lower-extremity subungual melanoma were identified. Of 53 cases with a final diagnosis of melanoma, 18 were initially misdiagnosed. Misdiagnoses included wart, callous, fungal disorder, foreign body, crusty lesion, sweat gland condition, blister, nonhealing wound, mole, keratoacanthoma, subungual hematoma, onychomycosis, ingrown toenail, and defective/infected toenail. Of the 18 misdiagnosed cases, 9 were clinically amelanotic. CONCLUSION: Awareness that amelanotic variants of acral melanoma may assume the morphology of benign hyperkeratotic dermatoses may increase the rate of correct diagnosis and improve patient outcome. PMID- 12582387 TI - Tinea capitis in Cleveland: survey of elementary school students. AB - BACKGROUND: Tinea capitis, a fungal infection of the scalp, is of increasing public health importance, and Trichophyton tonsurans has become the primary causative agent in North America. OBJECTIVES: To determine the prevalence of dermatophyte-positive scalp cultures among elementary schoolchildren in Cleveland, Ohio, describe predisposing factors, and measure the antifungal susceptibility of isolates collected. OBSERVATIONS: A total of 937 children from 8 Cleveland elementary schools were cultured for the presence of dermatophytes; 122 children (13%), all of whom were African American, had dermatophyte-positive cultures of the scalp. Sixty percent of cases were asymptomatic, indicating a carrier state. Race, scaling, and the use of anti-dandruff shampoo were associated with increased likelihood of infection. T tonsurans was the only organism isolated (except 1 Microsporum canis isolate). All isolates were susceptible to fluconazole, griseofulvin, itraconazole, and terbinafine. CONCLUSIONS: T tonsurans was the predominant dermatophyte isolated. Further multicenter studies are needed to confirm the predominance of dermatophyte positive scalp cultures among African American children and to determine modifiable and preventable risk factors. PMID- 12582388 TI - Deodorants: an experimental provocation study with cinnamic aldehyde. AB - BACKGROUND: Axillary dermatitis is common and overrepresented in individuals with contact allergy to fragrances. Many individuals suspect their deodorants to be the incriminating products. OBJECTIVE: Our aim was to investigate the significance of cinnamic aldehyde in deodorants for the development of axillary dermatitis when used by individuals with and without contact allergy to cinnamic aldehyde. METHODS: Patch tests with deodorants and ethanol solutions with cinnamic aldehyde, and repeated open application tests with roll-on deodorants without and with cinnamic aldehyde at different concentrations, were performed in 37 patients with dermatitis, 20 without and 17 with contact allergy to cinnamic aldehyde. RESULTS: A repeated open application test with positive findings was noted only in patients hypersensitive to cinnamic aldehyde (P <.001) and only in the axilla to which the deodorants containing cinnamic aldehyde had been applied (P <.001). CONCLUSION: Deodorants containing cinnamic aldehyde in the concentration range 0.01% to 0.32%, used twice daily on healthy skin, can elicit axillary dermatitis within a few weeks. PMID- 12582389 TI - Adverse cutaneous reactions to imatinib (STI571) in Philadelphia chromosome positive leukemias: a prospective study of 54 patients. AB - BACKGROUND: Imatinib is a new major treatment in chronic myeloid leukemia. OBJECTIVE: To study the cutaneous reactions induced by imatinib. METHODS: All inpatients and outpatients with Philadelphia chromosome-positive leukemia treated by imatinib were included in this prospective study. Clinical features, pathologic findings, evolution of each case, and analysis of potential risk factors were recorded. RESULTS: A total of 54 patients were included, 48 of whom experienced at least 1 cutaneous reaction. These reactions consisted of 36 rashes, 35 edemas, and 22 pruritus. The rash was severe in 5 patients, resulting in temporary interruption of treatment in 3. Highly significant relationships were observed between the daily dose of imatinib and both rashes and edema. In a multivariate analysis, female sex and the daily dose of imatinib were independent risk factors for the development of rashes. CONCLUSION: Adverse cutaneous reactions induced by imatinib are frequent, generally moderate, and dose dependent. PMID- 12582390 TI - Organotypic cultures of autologous hair follicle keratinocytes for the treatment of recurrent leg ulcers. AB - Our purpose was to evaluate, in an open study, the efficacy of epidermal equivalents (EEs), a tissue-engineered epidermis prepared from autologous hair follicle keratinocytes, for the treatment of recurrent leg ulcers (n = 50). To generate EEs, keratinocytes expanded from the outer root sheaths of plucked anagen hair follicles were seeded on cell culture inserts at air-liquid interface. The total culture time was 5 to 6 weeks. Three days after the procedure, 95% of EEs adhered to the wound bed. After 8 weeks, 70% of the total wound surface was re-epithelialized and 32% of the ulcers were healed. After applying the EEs, a major relief of wound pain was noticed by the patients. EEs were applied in ambulatory patients without surgical facilities. Because 92% of the cases included in this study presented a recurrence of their ulcers after a split-thickness skin graft, we consider these ulcers as difficult to treat and propose the EEs as an alternative effective treatment of recurrent leg ulcers. PMID- 12582391 TI - Narrowband UVB and psoralen-UVA in the treatment of early-stage mycosis fungoides: a retrospective study. AB - BACKGROUND: Treatment of early-stage mycosis fungoides (MF) consists of topical steroids, phototherapy (UVB), photochemotherapy (psoralen plus UVA [PUVA]), topical nitrogen mustard, or total skin electron-beam irradiation. It has been reported that the same effective UVB dose is safer than PUVA regarding carcinogenicity and produces fewer side effects. Narrowband UVB (311 nm) results in less irritation and erythema and is more effective compared with broadband UVB. OBJECTIVE: Our purpose in this retrospective study was to analyze the response to treatment, relapse-free interval, and irradiation dose in 56 patients with early-stage MF (stage Ia and Ib). A total of 21 patients were treated with narrowband UVB (311 nm); 35 patients were treated with PUVA. RESULTS: Narrowband UVB treatment led to complete remission in 17 of 21 patients (81%), partial remission in 4 of 21 (19%), and none showed progressive disease. PUVA treatment led to complete remission in 25 of 35 patients (71%), partial remission in 10 of 35 (29%), and none showed progressive disease. The mean relapse-free interval for patients treated with UVB was 24.5 months (range, 2-66 months) and for patients treated with PUVA, 22.8 months (range, 1-43 months). CONCLUSION: Narrowband UVB therapy for patients with early-stage MF is an effective treatment modality. It has several advantages over treatment with broadband UVB and PUVA. When treating patients with early-stage MF it may be beneficial to start with narrowband UVB therapy and, if there is progression or no response, switch to PUVA therapy. PMID- 12582392 TI - Sezary syndrome and seronegative polyarthritis: treatment with extracorporeal photochemotherapy. AB - We describe a patient with therapy-resistant cutaneous T-cell lymphoma, Sezary syndrome variant, in association with concurrent polyarthritis and vitiligo, who was successfully treated with extracorporeal photochemotherapy (ECP). The combination of Sezary syndrome with seronegative rheumatoid arthritis is rare. In our patient the T-cell lymphoma was refractory to standard treatments that included psoralen-UVA, lymph node irradiation, and polychemotherapy. ECP has been shown to be effective in the treatment of selected cases of Sezary syndrome. There is a strong suggestion that ECP as a monotherapy can provide a significant benefit for other T-cell-mediated diseases including rheumatoid arthritis. In spite of a disease duration of 10 years, a very low CD8 cell count (2% of lymphocytes), a very high CD4 cell count (94%), and multiple unsuccessful chemotherapeutic trials before initiation of ECP, our patient achieved a long lasting complete remission of both diseases with normalization of the CD4+ and CD8+ T-lymphocyte subsets. Concurrent developing vitiligo was unaffected by ECP. PMID- 12582393 TI - Photodynamic therapy with topical methyl aminolevulinate for actinic keratosis: results of a prospective randomized multicenter trial. AB - BACKGROUND: Photodynamic therapy (PDT) is a promising new treatment modality for actinic keratoses. Methyl aminolevulinate (MAL) (Metvix, PhotoCure, Oslo, Norway) leads to selective accumulation of photoactive porphyrins in premalignant skin lesions and makes the lesions susceptible to phototoxic effects on illumination with red light. OBJECTIVE: This multicenter, randomized, double-blind study compared complete response rates, cosmetic outcome, and patient satisfaction for PDT with cream containing 160 mg/g MAL or placebo cream in the treatment of actinic keratoses. METHODS: After application of the cream under occlusion for 3 hours, the lesions were illuminated by noncoherent red light (570-670 nm, light dose 75 J/cm(2)). Treatment was repeated after 1 week and response was assessed 3 months later. A total of 80 patients were randomized into the study, 42 in the active and 38 in the placebo group. RESULTS: Complete lesion response rate was higher after MAL PDT than placebo, 89% versus 38% per protocol analysis (P =.001). An excellent or good cosmetic outcome was reported in more than 90% of patients treated with MAL. CONCLUSION: In this small study, PDT using topical MAL was a safe and effective treatment for actinic keratoses with excellent cosmetic outcome. It is a promising treatment that could benefit from further study. PMID- 12582394 TI - Thrombotic complications related to discontinuation of warfarin and aspirin therapy perioperatively for cutaneous operation. AB - BACKGROUND: Aside from anecdotal reports, there are few data on the risk of thrombotic complications in patients in whom use of warfarin and aspirin is discontinued perioperatively for cutaneous operation. OBJECTIVE: Our aim was to present a large case series of thrombotic complications resulting from this practice and to estimate the incidence of these events. METHODS: A total of 504 members of the American College of Mohs Micrographic Surgery and Cutaneous Oncology were surveyed regarding thrombotic complications when blood thinners were withheld perioperatively to ascertain the frequency of these complications and to describe associated morbidity and mortality. RESULTS: A total of 168 responding physicians reported 46 patients who experienced thrombotic events. Of these patients, 54% (25 of 46) experienced the event when warfarin was withheld and 39% (18 of 46) when aspirin use was discontinued. Thrombotic events included 24 strokes, 3 cerebral emboli, 5 myocardial infarctions, 8 transient ischemic attacks, 3 deep venous thromboses, 2 pulmonary emboli, and 1 retinal artery occlusion leading to blindness. Three deaths were reported. Calculation of incidence yielded an estimated thrombotic risk of 1 event per 12,816 operations, 1 in 6219 operations when use of warfarin was discontinued and 1 in 21,448 when aspirin was withheld. CONCLUSIONS: With no documented increase in severe hemorrhagic complications during continued use perioperatively of blood thinners, these data provide a compelling argument to maintain patients on medically necessary blood thinners during cutaneous operation. All relevant clinical facts must be weighed when making this decision. PMID- 12582395 TI - Folliculotropic mycosis fungoides with central nervous system involvement: demonstration of tumor clonality in intrafollicular T cells using laser capture microdissection. AB - BACKGROUND: Folliculotropic mycosis fungoides (MF) is a rare variant of cutaneous T-cell lymphoma, MF type, characterized by atypical lymphocytes preferentially infiltrating the hair-follicle epithelium relative to the epidermis. OBSERVATIONS: We describe a rare case of folliculotropic MF involving the central nervous system. This is also the first case in which laser capture microdissection was used to show that the atypical lymphocytes within the hair follicle epithelium were part of the same tumor clone present in other tissue compartments. CONCLUSIONS: In reviewing the literature describing atypical lymphocytes infiltrating hair-follicle epithelium relative to the epidermis, we encourage the use of the term folliculotropic mycosis fungoides. Our case also supports previous findings that central nervous system involvement can occur in advanced MF. The successful procurement and analysis of atypical lymphocytes from hair-follicle epithelium by laser capture microscopy ushers in a new era in molecular diagnostics. PMID- 12582396 TI - Desmoglein as a target in autoimmunity and infection. AB - Clinical phenotypes of most diseases are complex. However, once the mechanism behind the scene is clarified, the nature shows amazing beauty. There is a simple logic behind a complex disease. The exact molecular mechanism of the blister formation in staphylococcal scalded skin syndrome (SSSS) remained to be elucidated for 3 decades since exfoliative toxin was discovered by Melish and Glasgow in 1970. A knowledge accumulated to understand the pathogenesis of pemphigus and cell-cell adhesion of keratinocytes led us to solve this question. Desmoglein 1, which is a cadherin type cell-cell adhesion molecule in desmosomes, is targeted in two different skin diseases, pemphigus foliaceus, and SSSS. In pemphigus foliaceus IgG autoantibodies are developed against desmoglein 1 and inhibit its adhesive function with resultant blister formation in the superficial epidermis. In SSSS, exfoliative toxin produced by Staphylococcus aureus specifically binds and cleaves desmoglein 1 with resultant blister formation at the identical site. PMID- 12582397 TI - Current and novel methods for assessing efficacy of hair growth promoters in pattern hair loss. PMID- 12582398 TI - Papillary tip bleeding or the Auspitz phenomenon: a hero wrongly credited and a misnomer resolved. PMID- 12582399 TI - Dermatology: a unique case of specialty workforce economics. AB - Physician workforce economics in the United States have changed substantially over the past few years as a result of oversupply in many specialties and the emergence of managed care as the predominant health care delivery system. Some newly graduating residents have reported difficulty in finding jobs, unsatisfactory compensation levels, and low job satisfaction. However, dermatology as a specialty has experienced surprisingly prosperous employment opportunities. Recent studies have affirmed that recent dermatology residency graduates have had less difficulty than their colleagues in securing desirable positions. This article examines 4 different factors that may have set dermatology apart from the rest of the medical specialties: the supply of new dermatologists, the impact of managed care and shifting health care systems, the impact of changing professional demographics, and the changing scope of practice. Trends in these 4 areas have tightened the supply and increased the demand for dermatologists. Whether future residents will enjoy a similarly open market remains to be seen as the US health care system evolves. PMID- 12582400 TI - Surgical pearl: Removal of cosmetic lip-liner tattoo with the pulsed carbon dioxide laser. PMID- 12582401 TI - Biopsy technique pearl: Obtaining an optimal split punch-biopsy specimen. PMID- 12582402 TI - A complete and durable response to denileukin diftitox in a patient with mycosis fungoides. AB - We describe a patient with stage IA mycosis fungoides treated with denileukin diftitox as part of a randomized phase III trial. The patient had a rapid and complete response and has remained in remission for more than 5 years without maintenance therapy. Although denileukin diftitox is not usually considered for patients with early-stage mycosis fungoides, our experience provides anecdotal evidence that fusion toxin therapy may be worth considering as a treatment for patients with stage I disease. PMID- 12582403 TI - Basal cell carcinoma of the toenail unit. PMID- 12582404 TI - Scleroderma-like cutaneous lesions induced by paclitaxel: a case study. AB - Paclitaxel is a recent antineoplastic agent that belongs to the taxane family. Its activity has been demonstrated in advanced and refractory ovarian, breast, lung, and head and neck cancer. Adverse cutaneous reactions to paclitaxel have been reported, namely bullous fixed drug eruption, onycholysis, acral erythema, erythema multiforme, and pustular eruption. We report the first case of scleroderma-like changes after paclitaxel administration. A 63-year-old patient presented with an edematous and infiltrated erythema of the head, neck, axillae, and left hand 10 days after administration of paclitaxel and paraplatin for primitive peritoneal cancer. Cutaneous lesions improved after a change from paclitaxel to cyclophosphamide. Cutaneous lesions recurred 3 months later, after reintroduction of paclitaxel, and progressively evolved to cutaneous sclerosis. Skin biopsy showed a dermal fibrosis. Biologic tests revealed no autoimmunity. Scleroderma-like lesions of this patient were reminiscent of previously reported cases that occurred after administration of docetaxel, which also belongs to the taxan family. Thus, scleroderma-like syndromes seem to represent a unique cutaneous adverse event caused by taxanes. PMID- 12582405 TI - A case of multiple keratoacanthoma centrifugum marginatum: response to oral etretinate. AB - The patient was a 48-year-old man who had a history of colonic polyps. About 18 months before, a dark reddish nodule had appeared on his right forearm. By peripheral growth, this lesion had formed a crateriform prominence with central healing. Similar symptoms multiplied and expanded, the largest being about the size of a child's head. Given both the clinical and histologic evidence, a diagnosis of keratoacanthoma centrifugum marginatum (KCM) was made. Oral etretinate was administered, and an involution of verrucous plaque began after 4 weeks. After 8 weeks, the lesions faded, leaving pigmentation in some areas. This is a rare disorder, only 30 such cases having been reported worldwide, and the present case was the third in which oral etretinate was found to be effective. PMID- 12582406 TI - Rickettsialpox in a patient with HIV infection. AB - We describe the first case of rickettsialpox in a patient infected with HIV. Immunohistochemical staining of biopsied lesions showed a relatively large number of rickettsiae within the papulovesicular rash. Rickettsialpox is easily treated and may resemble more serious cutaneous eruptions in patients infected with HIV. This diagnosis should be considered in immunocompromised city-dwellers, with fever and a papulovesicular rash. PMID- 12582408 TI - Allergic contact dermatitis from topical doxepin: Food and Drug Administration's postmarketing surveillance experience. AB - A total of 26 postmarketing cases of allergic contact dermatitis to doxepin 5% cream were reported to the Food and Drug Administration. Our findings suggest that allergic contact dermatitis was more common when treatment duration exceeded the recommended 8 days. Allergic contact dermatitis to doxepin cream should be considered in patients whose condition fails to improve or worsens with doxepin use. PMID- 12582407 TI - Progressive cutaneous sarcoidosis responding to anti-tumor necrosis factor-alpha therapy. AB - We describe a patient with widespread and progressive cutaneous sarcoidosis who was successfully treated with anti-tumor necrosis factor-alpha monoclonal antibody, infliximab, suggesting that inhibition of tumor necrosis factor-alpha may be useful as targeted treatment in cutaneous sarcoidosis. PMID- 12582409 TI - Glucagonoma syndrome: diagnosis and treatment. PMID- 12582410 TI - Arcanobacterium haemolyticum. PMID- 12582411 TI - Acetaminophen-induced linear IgA bullous dermatosis. PMID- 12582412 TI - Black henna tattoo reaction in a person with sulfonamide and benzocaine drug allergies. PMID- 12582413 TI - Intravenous regional anesthesia (Bier's block) is superior to a peripheral nerve block for painless treatment of plantar hyperhidrosis with botulinum toxin. PMID- 12582414 TI - Evidence-based evaluation of photodynamic therapy of actinic keratoses. PMID- 12582415 TI - Isotretinoin (Accutane) and serious psychiatric adverse events. PMID- 12582416 TI - Dapsone and sulfones in dermatology: overview and update. PMID- 12582417 TI - Journalology and citation etiquette: firstedness might matter, but only if you're really first! PMID- 12582418 TI - The work has just begun for urologists. Highlights from the 97th annual meeting of the American Urological Association, Orlando, Florida, May 25-30, 2002. PMID- 12582419 TI - Oral lichen planus: an update. AB - Oral lichen planus (OLP) is a chronic autoimmune disease of unknown etiology that affects the skin and mucosae, including the oral cavity. The disease is characterized by lacy, thin white lines on a violaceous background on the oral mucosa, usually on the inside of the cheeks. The disease is also characterized by chronic inflammation and is often associated with severe pain and a burning sensation in the mouth. The etiopathogenesis of lichen planus is complex, with the involvement of T lymphocytes, mast cells, intercellular adhesion molecule-1 (ICAM-1) and major histocompatibility complex class II antigens. The immunologic process results in vacuolar degeneration, lysis of basal cells and, ultimately, liquefaction of the basal cells. The precipitating factors of OLP can be: stress, particular foods, dental plaque, systemic illness and poor oral hygiene. Often no medication is necessary for benign disease. In the case of severe pain and the burning sensation, high potency topical corticosteroids remain the most reliably effective treatment. Other available treatments are topical cyclosporine, tacrolimus, retinoids and other immunosuppressive agents. Systemic corticosteroids may be indicated in patients whose condition is unresponsive to topical corticosteroids. However, OLP remains a challenging disease to treat, in spite of the numerous treatments tried in the patient population. PMID- 12582420 TI - A six-month trial of valacyclovir in the Epstein-Barr virus subset of chronic fatigue syndrome: improvement in left ventricular function. AB - This study was designed to determine safety and efficacy of a 6-month trial of valacyclovir in single-virus Epstein-Barr virus (EBV) persistent infection. Phase I of this study used four specific criteria to define a subset of patients with chronic fatigue syndrome (CFS). In the second phase, myocardial dynamics were measured by MUGA rest/stress radionuclide ventriculographic (RVG) examinations pre- and posttreatment with valacyclovir. In phase I, a trial was performed in 19 consecutive CFS patients with the following diagnostic conditions: patients met criteria for diagnosis of CFS; they had had CFS for less than 1 year. They demonstrated repetitively abnormal oscillating T waves (ischemic or flat) at 24-h Holter monitoring; and they had elevated serum IgM antibody titers to EBV viral capsid antigen and/or total diffuse early antigen as measured by the enzyme linked immunosorbent assay method. The treatment group comprised 10 CFS patients with no serum antibodies to human cytomegalovirus, but the control group (nine CFS patients) had, additionally, high titers of serum antibodies (IgG) to conformational structural antigens of human cytomegalovirus. Both the parallel treatment and control CFS groups received valacyclovir 1.0-1.5 gm q.6.h. for 6 months. This valacyclovir dose achieved serum acyclovir C(max) of > 7 microm and high antiviral activity versus EBV (IC(50) of 4.4-13.3 m). In phase II, six additional CFS patients met the same four criteria as the 19 CFS patients in phase I. They had, however, been ill for a mean of 55.8 months. Thus, 25 CFS patients comprise this study. The studies were carried out at a single outpatient practice in Birmingham, MI, U.S.A. Before initiating valacyclovir, and after 6 months of treatment, clinical and laboratory observations were made. The CFS Energy Index point score (Table I) was used to record each CFS patient's functional capacity at baseline and after 1, 3 and 6 months of valacyclovir. Energy Index point scores, as well as EBV and human cytomegalovirus serum antibody titers were assessed. In the second phase, left ventricular dynamics were repeated after 6 months of treatment with valacyclovir. We concluded that the 16 CFS patients (included in both phases of this study) with EBV-persistent infection (EBV single-virus subset) are improved after 6 months of continuous pharmacokinetic dosing with valacyclovir. Nine CFS patients with EBV/human cytomegalovirus co-infection did not benefit from 6 months of similar treatment. Valacyclovir is not an effective anti-human cytomegalovirus antiviral drug. Unimproved CFS patients with co-infections EBV and human cytomegalovirus may require combined treatment with valacyclovir and another drug more active against human cytomegalovirus. This preliminary trial, with a small number of patients, may be critical to an appropriately designed larger, double-blind, placebo controlled trial. PMID- 12582421 TI - Medical backgrounders: glaucoma. AB - Glaucoma is a leading cause of irreversible blindness in the world. Currently, glaucoma is diagnosed as a progressive optic neuropathy with characteristic optic disc and nerve fiber layer damage, usually associated with loss of visual function. The intraocular pressure (IOP) is the most important risk factor for the disease, although a significant proportion of patients do not have elevated IOP. Other risk factors include older age, African descent, myopia and family history of the disease. The ophthalmoscopic examination of the optic disc is essential to identify the signs of glaucomatous optic neuropathy, such as increased cupping, neuroretinal rim thinning or optic disc hemorrhages. Glaucomatous visual field loss usually starts in the periphery, and loss of central vision does not occur until late in the course of the disease. Visual function is most commonly assessed by standard automated perimetry; however, as many as 50% of nerve fibers can be lost before the appearance of visual field defects in this test. Newer technologies have been developed to find more sensitive ways to detect early glaucoma using both functional (short-wavelength automated perimetry and frequency-doubling perimetry) and structural (scanning laser topography, optical coherence tomography and scanning laser polarimetry) measurements. The management of glaucoma is based on lowering the intraocular pressure to prevent further optic nerve damage. Currently, there are five major classes of medications that are used to lower the intraocular pressure: Beta adrenergic antagonists, adrenergic agonists, parasympathomimetics, prostaglandin like analogues and carbonic anhydrase inhibitors. The goal of therapy is to maintain adequate vision for patients during their lifetime, keeping in mind the possible adverse effects of the drugs. If additional lowering of IOP is indicated or if medication fails to sufficiently lower the IOP, laser trabeculoplasty is usually the next step. If IOP is still not adequately controlled, incisional glaucoma surgery is indicated. Neuroprotective agents, which directly protect the optic nerve in glaucoma, are being evaluated in clinical trials. PMID- 12582422 TI - The basis for the chemopreventive action of resveratrol. AB - The polyphenolic phytoalexin resveratrol (3,5,4'- trihydroxy-trans-stilbene) is produced, perhaps primarily, as a natural fungicide by more than 70 plant species, and can be found in high to moderate quantities in various foods including grapes, peanuts and wine. Recent in vitro and a limited number of in vivo studies have documented that physiological concentrations of resveratrol can modulate multiple molecular pathways thought to be associated with the development and progression of cardiovascular disease and cancer, among them phase II drug metabolizing, cyclooxygenase, lipid metabolizing, nitric oxide, DNA synthesis, inflammation, cell survival, cell death and cell division cycle pathways. Work on the mechanisms underlying the cytostatic activities, which appear to affect all dividing tissues, and the cytotoxic activities of the compound, which seem to preferentially target tumor cells, has produced some controversial and, at times, seemingly conflicting results. The present review attempts to integrate some of the established biochemical activities of resveratrol into a common framework of function in an attempt to understand precisely how the compound affects cell proliferation and survival. PMID- 12582423 TI - [Trans-gram gene transfer: the case of beta-lactamases]. AB - Lateral or horizontal gene transfer is a phenomenon that has influenced the evolution of microorganisms. Despite the evolutionary trend toward genetic isolation, lateral transfer seems to take place relatively frequently, bridging the gap between very separate species. The acquisition of foreign genes may have accelerated in recent years because of the increase in the adaptive needs of bacteria, particularly through the use of antibiotics. Transfer of genes encoding beta-lactamases from Gram-positive to Gram-negative bacteria (trans-gram transfer) may be suggested on the basis of sequence analysis. We found that the sequences of the beta-lactamases BRO-1 and ACl-1, from the Gram-negative bacterial organisms Moraxella and Acidaminococcus, respectively are abnormally placed among sequences from Gram-positive beta-lactamases in phylogenetic trees. In both cases, the topology of the enzyme (attached to the cellular membrane), the structure of the signal peptide, and the Shine-Dalgarno region suggest that these enzymes originated from Gram-positive organisms. Results remain inconclusive for Haemophilus ROB-1 beta-lactamase. PMID- 12582424 TI - [Staphylococcus aureus resistance. Now vancomycin]. PMID- 12582425 TI - [Present perspectives on Creutzfeldt-Jakob disease: pathogenesis, diagnosis and treatment]. AB - In recent years, interest in the group of diseases generically known as "transmissible spongiform encephalopathies" has reemerged, mainly due to the socioeconomic implications of one of its variants: bovine spongiform encephalopathy. Transmissible spongiform encephalopathies seem to be caused by proteinaceous particles called prions, isoforms of a membrane protein which can be found in humans as well as in several animal species. Their precise function is still unknown. Although investigation into transmissible spongiform encephalopathies has clarified some uncertainties about its pathogenesis and epidemiology, many important questions remain. This is especially true or treatment, since no effective curative therapy exists as yet. PMID- 12582426 TI - [Influence of the application of a rational, consensual protocol on the use of antibiotics in a general hospital]. AB - A large increase in the use of wide-spectrum antibiotics has occurred in Spain. The objective of our study was to evaluate the influence of applying protocols for the use of such antibiotics. A prospective, observational study was carried out based on the information provided by the pharmacy service about prescriptions of ceftriaxone, cefotaxime, ceftazidime, imipenem and ciprofloxacin at the single dosing department of the hospital. Two groups were included in the study: 1) control group (October-December 1995), which did not take into account the protocols for antibiotic treatment; and 2) the study group, during the same months in 1996, which took into account the consensual protocols. The criteria for the type of patient, infection and outcome, and type of antibiotic treatment were evaluated according to the rules of the international literature for this type of study. The baseline characteristics of both groups of patients were similar. Following the application of the protocol, a decrease in the use of antibiotics was observed (108 cases in group 2 vs. 207 cases in group 1); an increase in the appropriateness of treatment (50.72% in group vs. 77.57% in group 2); an increase in the cure rate (78.26% in group 1 vs. 91.67% in group 2); and a decrease of 5.01 days in the length of hospital stay (22.99 days in group 1 vs. 17.98 days in group 2), which was statistically significant, and thus led to a savings of 2073.74 Euros per cured patient. The application and adherence to a consensual protocol is a beneficial method of using antibiotics since it improves the quality of their use and makes it more cost-effective. PMID- 12582427 TI - [Epidemiological situation of tuberculosis in the province of Salamanca and the influence of HIV/AIDS infection]. AB - This study aimed to determine the real prevalence of tuberculosis and the influence of HIV/AIDS infection from January 1992 to January 2001 in the health area of Salamanca. Data on tuberculosis were obtained from the EDO system of the Medical Records Service and the Microbiology Department of the University Hospital of Salamanca. Data on HIV/AIDS infection were obtained from the records on seropositive patients from the Preventive Medical Service. It was found that during the study period, 769 cases of tuberculosis were diagnosed and 606 cases were reported, 12.7% of which were in HIV/AIDS patients. There was 8.7% resistance to isoniazid and 8.3% to rifampin. Multidrug resistance was found in 4.17%. It was concluded that there is a close relationship between tuberculosis and HIV/AIDS infection, which may constitute a risk factor for the disease as well as for the appearance of multidrug resistance. The low reporting of tuberculosis cases shows the need for active surveillance systems. PMID- 12582428 TI - [Retreatment with interferon plus ribavirin in chronic hepatitis C patients with genotype 1 who had no response or a relapse with a previous interferon treatment. Long term follow-up]. AB - The aims of this study were twofold: i) to evaluate the sustained response rate at 6 months, at 12 months and at the end of the follow-up [(46.39 +/- 9.74 months) (range: 6-60 months)] in patients with chronic hepatitis C retreated with interferon-a (IFN-a) plus ribavirin for 6 months (group A), 12 months (group B), in patients with relapse and those with no response to a previous course of monotherapy with IFN-a; and ii) to evaluate changes in the histological liver lesion. One hundred and four patients (100 with genotype 1 and four without), 44 with relapse and 60 non-responders, were included. A total of 20.2% of the patients were excluded because of side effects. Fifty percent of the relapsing patients with genotype 1 showed a sustained response. The sustained response rate was higher in group B than in group A, whereas the rate for the relapsing patients was lower in group B than in group A (13/20 vs. 5/16; 7.2% vs. 61.5%; p <0.09 and p <0.009, respectively). However, 9.3% of the non-responding patients showed a sustained response (A/B: 2/2). The relapsing patients who had a genotype other than 1 showed a sustained response. The sustained response rate was constant throughout the follow-up, except in relapsing patients from group A (the late relapse rate was 28.6%). The viral load was not related to the response. The grade and stage of the histological lesion improved in patients with sustained response (n=20) and in one-third of the non-responding patients (n=45) (p <0.0001 and p <0.0001). IFN in combination with ribavirin was more effective in relapsing patients with genotype 1 at 12 months than in those at 6 months of treatment, whereas the effectiveness in non-responding patients was low. The sustained response evaluation at 6 months was reliable, except in relapsing patients with 6 months of treatment. The histological lesion improved significantly with combination antiviral therapy. PMID- 12582429 TI - Immediate vs. delayed imipenem treatment in cancer patients with profound neutropenia induced by high-dose chemotherapy: results of a randomized study. AB - A study was carried out to compare the use of prophylactic imipenem administered at the onset of profound neutropenia (immediate) with therapeutic imipenem administered at the onset of neutropenic fever (delayed) in cancer patients treated with high-dose chemotherapy. A total of 65 patients who were scheduled to receive two cycles of high-dose cyclophosphamide, etoposide, cisplatin (CEP) chemotherapy were randomized to receive imipenem either at presentation of neutropenia (immediate imipenem arm, prophylactic arm) or at commencement of neutropenic fever (delayed imipenem arm, therapeutic arm). Treatment was crossed over when the second CEP chemotherapy cycle was received. Of the 65 patients, 41 received the two planned cycles and 24 received only the first. Compared with the delayed imipenem arm, the immediate imipenem arm was associated with lower fever incidence (86.3% vs. 100%, p=0.0142) and Gram-negative bacteria infection [4/51 (7.8%) vs. 14/55 (25.5%), OR=0.24, p =0.031]. There were fewer episodes of pneumonia (2% vs. 12.7%), septic shock (0% vs. 3.6%) and deaths from infection (0% vs. 3.6%), but these differences did not reach statistical significance. With regard to delayed imipenem, for every seven patients with immediate imipenem, one episode of febrile neutropenia was avoided; for every six patients administered immediate imipenem, one case of Gram-negative infection was avoided; and for every nine patients administered immediate imipenem, one episode of pneumonia was avoided. There were no differences in the incidence of Gram-positive infections nor in the length of hospitalization between the two treatment arms. In conclusion, compared to its conventional delayed use, immediate imipenem significantly reduces the frequency of febrile neutropenia and Gram-negative infections in patients with high-dose chemotherapy. PMID- 12582430 TI - [Methicillin-resistant Staphylococcus aureus. Present prevalence in an area in southern Spain]. AB - Methicillin-resistant Staphylococcus aureus (MRSA) poses a serious problem in some geographic areas. Its prevalence can change according to area and can even vary within a country. It is important to know the prevalence of resistance in each area so that the correct empirical treatment can be applied when these infections are suspected. We studied all the consecutive S. aureus isolates collected in our hospital from 2000-2001 and included only one isolate per patient. Identification of the isolate and determination of susceptibility were carried out using an automated system (VITEK, bioMerieux). High resistance to oxacillin was identified and high resistance was observed for erythromycin (41.22%), amoxicillin-clavulanic acid (26.48%), and ciprofloxacin (25.74%). S. aureus resistance to oxacillin was found to be 67.64% in catheter samples and 44.11% in urine samples. Multiresistant MRSA (to more than five antibiotics) accounted for 66.6%, with 4% resistant to two antibiotics. MRSA is a serious problem in our area and its prevalence is on the rise, as is multiresistance. The means of monitoring and controlling these strains need to be increased. PMID- 12582431 TI - [Study of the susceptibility of black yeasts of the Exophiala genus to antifungals using the Sensititre System]. AB - Black yeasts belong to a group of dematiaceous fungi which are difficult to classify and identify. One of the genera that cause human infection, mainly phaeohyphomycosis, is Exophiala, which includes the species E. berger, E. castellanii, E. dermatitidis, E. jeanselmei, E. lecanii-corni, E. moniliae, E. pisciphila, E. salmonis and E. spinifera. We carried out a susceptibility study of five of these species (E. castellanii, E.dermatitidis, E, jeanselmei, E. lecanii-corni and E. moniliae), isolated from cutaneous exudates, to the usual antimycotic agents. Strains were cultured on Sabouraud agar and potato-dextrose agar. Species identification was made by conidiogenous microscopic observation and the following characteristics: maximum growth temperature, tolerance to cicloheximide and carbohydrate assimilation. Susceptibility to antimycotic agents (amphotericin B, fluconazole, itraconazole, ketoconazole and 5-fluorocytosine) was determined by the Sensititre YeastOne (Aamar Blue) commercial method. Amphotericin B activity was good for the five tested species, although the MIC for E. jeanselmei was high (1 mg/l. MICs for fluconazole were high for all the species, but only E. moniliae and E. lecanii-corni c be considered resistant. Activity of itraconazole, ketoconazole and 5-fluorocytosine was good. PMID- 12582432 TI - Bacteriocin production by clinical Staphylococcus isolates. PMID- 12582433 TI - [Pneumonia due to Legionella. Macrolides or quinolones?]. PMID- 12582434 TI - [Mechanisms of bacterial resistance to quinolones]. AB - In order to produce their cytotoxic effect, quinolones must enter the cell through the bacterial membrane to reach their target, DNA-gyrase or topoisomerase IV, and induce cell death. The mechanisms of resistance to fluoroquinolones include: those mediated by gene mutations codifying for DNA gyrase and topoisomerase IV and leading to QRDR; those characterized by changes in the permeability of the outer membrane which decrease intracellular penetration of the drug; and those caused by active endogenous carriers responsible for drug efflux. These resistance mechanisms can occur alone or in combination; in fact, it is believed that high resistance levels to quinolones in vivo are associated with simultaneous mechanisms. This article reviews such resistance mechanisms and establishes, when possible, their relation to the structure of quinolones. PMID- 12582435 TI - [Correlation between in vitro susceptibility to antifungal drugs and the clinical evolution of patients with candidiasis and cryptococcosis]. AB - The increase in the incidence of fungal infections and the emergence of resistance call for the development of techniques for measuring in vitro antifungal susceptibility that are useful for predicting clinical outcome in patients suffering from these infections. In the past, the lack of standardized testing techniques led to poor intra- and interlaboratory reproducibility. Recently, the National Committee for Clinical Laboratory Standards (NCCLS) has developed a reference method for antifungal susceptibility testing, document M27A. This document is a necessary and important step towards the standardization of antifungal susceptibility testing, which has important implications in the analysis of clinical and microbiological data. This article provides a comprehensive review of studies correlating in vitro antifungal susceptibility testing and clinical outcome. In general, it is possible to predict the therapeutic outcome, especially in HIV infected patients with oropharyngeal candidiasis treated with fluconazole. However, in other more heterogeneous groups of patients it is more difficult to correlate the in vitro and in vivo data. PMID- 12582436 TI - [Comparison of three genotyping methods for the detection of HIV-1 resistance to antiretroviral drugs]. AB - Highly active antiretroviral therapy has dramatically improved the life expectancy of human immunodeficiency virus (HIV)-infected patients, but mutations in the HIV-1 reverse transcriptase (RT) and protease (P) genes confer drug failure. Evaluation of drug resistance genotyping in HIV-1 has proven to be useful for the selection of drug combinations with maximum antiretroviral activity. The aim of this study was to evaluate the optimal procedure to determine the resistance profile in the laboratory. Plasma from 90 antiretroviral treated patients was analyzed by reverse hybridization, which identifies the presence of wild-types or mutations at the 19 key codons for protease and RT regions, and was compared with two other methods of direct cDNA sequencing. A total of 408 mutations were detected by InnoLiPA HIV-1, (Line Probe Assay, Innogenetics, Belgium), 572 by TrueGene HIV-1 Genotyping System (Visible Genetics, Canada), and 721 by ViroSeq HIV-1 Genotyping System (Perkin Elmer/Applied Biosystems, California). Hybridization detected a significantly higher number of primary mutations which are associated with a high level of drug resistance (p <0.001). Hybridization also detected a higher number of mixtures of wild-type and mutant viruses. There was a good concordance among the three methods, although it was higher between the two sequencing methods. Sequencing determines a higher number of mutations, but hybridization better identifies primary mutations correlated with a high level of drug resistance. Hybridization is more suitable for detecting mixed populations and is easier to implement in clinical laboratories but does not eliminate the need for sequence analysis for detection of drug-resistant HIV. PMID- 12582437 TI - [Viral quasispecies and their implications in antiviral therapy]. AB - The aim of this study was to evaluate the relationship between the quasispecies in the HVR1 region of the hepatitis C virus and treatment evolution in order to determine whether genetic complexity is predictive of response to interferon therapy. The samples were analyzed by nested RT-PCR-mediated single-strand conformation polymorphism assay (SSCP). Twelve patients with chronic hepatitis C were studied and divided into three groups: three patients with sustained response, three patients with transient response and six nonresponders. The patients in the sustained response group showed a low genetic complexity pattern. By contrast, in three nonresponders and in one patient with transient response, the SSCP assay revealed a high complexity pattern. With regard to the remaining patients with transient response, new SSCP bands appeared, thereby modifying their genetic complexity pattern. Therefore, nonresponse to interferon treatment could be related to the presence of a high genetic complexity pattern, while the detection of a low genetic complexity pattern is necessary for a positive response to interferon therapy. Due to the limited number of patients involved in this study, it was not possible to predict the response to interferon based on the genetic complexity pattern. Larger studies are therefore required. PMID- 12582438 TI - [Serotypes and antibiotic susceptibility of the genus Salmonella in Elche, Spain]. AB - Salmonellosis is one of the main public health problems in developed countries. The most common serotype is Salmonella enteritidis, followed by S. typhimurium, S. hadar and S. virchow. Each serotype's sensitivity to antibiotics is different, but in general, there has been an increase in the number of isolates resistant to antibiotics. This is especially true for S. hadar and nalidixic acid, although we have not isolated any strains resistant to ciprofloxacin. Our study provides data that may help to control these infections and draws attention to the increase in resistance to certain antibiotics. This should give rise to the implementation of measures to protect public health and control the use of antibiotics, especially in animals. PMID- 12582440 TI - [Staphylococcus haemolyticus and bacteremia]. PMID- 12582439 TI - [Use of systemic antibiotics in Valladolid (1996-2000)]. AB - A longitudinal retrospective study was conducted to analyze the changes in the use of systemic antibiotics from 1996 to 2000 in the province of Valladolid, Spain. The use of systemic antibiotics was determined according to the defined daily dose and showed a progressive increase during the first four years of the study, followed by a large decrease in the year 2000. When examined by subgroup, wide spectrum penicillins accounted for almost 60% of the antibiotics consumed, and were therefore responsible for this change. Macrolides represented 16.4% of the consumption, which increased during the whole study period. Quinolones, with 9.3% of the total, showed a slow but steady increase. Cephalosporins accounted for 8.1%, with their use decreasing since 1998. It was concluded that wide spectrum penicillins were the most used systemic antibiotics in the study population, although a decreasing trend in their use and an increase in the use of macrolides and quinolones was observed. PMID- 12582441 TI - [Penetration of antibiotics into human polymorphonuclear leukocytes]. PMID- 12582442 TI - [In-hospital oral treatment of skin and soft tissue infections]. PMID- 12582443 TI - [Study of the betalactam resistance of Haemophilus influenzae conferred by the bla(ROB-1) gene]. AB - Given the lack of information on the resistance to betalactams conferred by the bla(ROB-1) gene in our area, we decided to undertake a study to determine if and how often it appears in our isolates. We analyzed the presence of the gene in 177 strains of Haemophilus influenzae and compared the results with those from the biochemical tests (nitrocefin). Sixty-three strains tested positive in the nitrocefin test (35.6%). The presence of the bla(ROB-1) gene was detected in seven of the 177 strains (3.9%). All the strains expressing the gene also tested positive in the nitrocefin test. Of the 63 strains that showed biochemical resistance (positive to nitrocefin), seven were found to be caused by the presence of the bla(ROB-1) gene. An epidemiological follow-up for the presence of this gene in our area is therefore advisable. PMID- 12582444 TI - [Antimicrobial resistance of clinical strains of Salmonella enterica isolated in Zaragoza]. AB - In order to identify any changes in the incidence of Salmonella enterica serotypes and their resistance to a variety of antimicrobial agents, we conducted a retrospective study of all the strains isolated from stool samples at Hospital Clinico Universitario Lozano Blesa in Zaragoza from 1997 to 2000. We observed an increase in the number of isolates of Salmonella and Campylobacter and a decrease in other enteropathogens. Enteritidis was the most frequently isolated serotype (55.2%), showing an increasing tendency (from 44.1% in 1997 to 60.6% in 2000). Hadar, glostrup and virchow showed the highest rate of resistance to nalidixic acid. Enteritidis also showed an important increase in resistance to nalidixic acid (from 17.6% in 1997 to 41.4% in 2000). Typhimurium showed the highest resistance levels to ampicillin, chloramphenicol and cotrimoxazole. No resistance to fluoroquinolones or to cefotaxime was detected, with the exception of 0.5% of the S. enteritidis strains, which showed resistance to fluoroquinolones. PMID- 12582445 TI - [Susceptibility of Staphylococcus aureus isolated from blood to 11 antimicrobial agents and a review of the literature]. AB - From November 1998 to February 2000, a total of 91 strains of Staphylococcus aureus isolated from blood were analyzed for their susceptibility to 11 antimicrobial agents using the E-test method. Fifty-two isolates were methicillin susceptible (MSSA) and 39 were oxacillin-resistant (MRSA). All the MSSA were susceptible to gentamicin, ciprofloxacin, vancomycin, teicoplanin, rifampicin, fusidic acid, quinupristin-dalfopristin and linezolid; 90% were susceptible to erythromycin and 83% to mupirocin. All the MRSA were susceptible to vancomycin, teicoplanin, rifampicin and linezolid; 95% were susceptible to quinupristin dalfopristin; and 92% to gentamicin, mupirocin, fusidic acid. None of the MRSA were susceptible to erythromycin or ciprofloxacin. The susceptibility of SARM to erythromycin and ciprofloxacin was low, while the susceptibility to the rest of the antimicrobial agents remained active. PMID- 12582446 TI - [Second multicenter study of the in vitro susceptibility to piperacillin tazobactam]. AB - A study was conducted in 1999 on the activity of piperacillin-tazobactam on 1349 strains collected in a Spanish multicenter study involving 10 hospitals. A total of 897 Gram-negative bacteria of the following genuses were collected: Escherichia coli (169), Klebsiella spp. (112), Pseudomonas spp. (122), Enterobacter spp. (92), Serratia spp. (44), Citrobacter spp. (37), Proteus spp. (114), Morganella spp. (49), Acinetobacter spp. (47), Stenotrophomonas spp. (47), Haemophilus spp. (45), Moraxella spp. (19). The Gram-positive bacteria included Staphylococcus aureus (116), Streptococcus pneumoniae (92), and Enterococcus spp. (114). Piperacillin-tazobactam activity against different anaerobic bacteria was also determined: Bacteroides spp. (88), Clostridium spp. (25), Prevotella spp. (5), Fusobacterium spp. (3) and Peptosteptococcus spp. (9). The piperacillin tazobactam combination was found to maintain its activity against the isolated microorganisms with regard to the previous study. PMID- 12582447 TI - Tiotropium bromide: a novel once-daily anticholinergic bronchodilator for the treatment of COPD. AB - Tiotropium bromide (Spiriva, BA679BR, Boehringer Ingelheim) is a novel inhaled, long-acting anticholinergic bronchodilator that is employed as a once-daily maintenance treatment for patients with chronic obstructive pulmonary disease (COPD). Like ipratropium bromide, tiotropium bromide is a quaternary ammonium derivative that binds to muscarinic receptors. However, although tiotropium binds with high affinity to muscarinic receptors of M1-, M2- and M3-subtypes, it dissociates very slowly from M1- and M3-receptors but more rapidly from M2 receptors, thereby giving it a unique kinetic selectivity. To date, the short acting anticholinergic agents ipratropium and oxitropium bromide have been extensively employed as bronchodilator therapy for patients with COPD. Indeed, the Global Initiative for Chronic Obstructive Lung Disease (GOLD) strategy emphasises the role of bronchodilators in symptomatic management of all stages of COPD. It is encouraging that tiotropium given once daily from a dry powder inhaler at 18 g has been shown to cause greater improvement in lung function and reduction in symptoms than ipratropium bromide given four times daily. Furthermore, clinical studies over a 1-year period have demonstrated that tiotropium has impressive and maintained effects on lung function, symptoms and health-related quality of life, and may also reduce exacerbations. In a recent large scale comparative study over 6 months, tiotropium has been shown to cause superior bronchodilation and symptomatic improvement when compared to twice daily salmeterol in COPD. The only significant reported adverse event is dry mouth, which is found in approximately 10%-15% of subjects, but this is reversible and rarely causes discontinuation of therapy. Based on these promising features, it is likely that tiotropium used alone or in combination with other bronchodilators will emerge as first-line maintenance treatment for patients with airway obstruction due to COPD. PMID- 12582448 TI - Imatinib mesylate (Gleevec, Glivec): a new therapy for chronic myeloid leukemia and other malignancies. AB - Imatinib mesylate (STI571, Gleevec, Glivec, a selective inhibitor of the BCR-ABL tyrosine kinase causative of chronic myeloid leukemia (CML), represents the paradigm of how a better understanding of the pathogenetic mechanisms of a neoplastic disease can lead to the development of a targeted molecular therapy. Phase II clinical trials have shown marked therapeutic activity of imatinib in all evolutive phases of CML, but notably in the chronic phase, where it induces complete hematological responses in almost 100% of patients resistant or intolerant to interferon, with a major cytogenetic response rate of 60%, including 41% complete cytogenetic responses. The preliminary results of an ongoing phase III multicenter randomized study comparing imatinib with interferon plus cytarabine as first-line treatment for CML favor imatinib in terms of efficacy and safety. If confirmed with longer follow-up,these results would establish imatinib as the choice therapy for the majority of CML patients, with allogeneic transplantation being restricted as initial therapy only to younger patients with a family donor. Longer follow-up will answer some questions, such as those on long-term safety, durability of the responses, whether these will translate into a survival prolongation and the possibility of molecular responses. In addition, further information on the mechanisms involved in the primary and acquired resistance to imatinib is needed. Besides the Bcr-Abl protein, the drug is also active against other tyrosine kinases, such as Abl, the stem-cell factor receptor (c-kit) and the platelet-derived growth factor receptor, whose inhibition might have potential implications for the treatment of several malignancies. In this sense, it must be pointed out that imatinib has shown a remarkable activity in gastrointestinal stromal tumors. PMID- 12582449 TI - Frovatriptan: a selective type 1B/1D serotonin receptor agonist for the treatment of migraine headache. AB - Frovatriptan belongs to an innovative family of compounds aimed at breaking through the long-standing barrier of migraine headache understanding and treatment. While a typology of headaches has been recognized for some time, and a number of therapies have been introduced for reduction of headache pain and duration, the causes of migraine remain a subject of debate. Those prone to attacks continue to endure them and suffer the related symptoms such as nausea and disorientation. Frovatriptan, like all the triptans, acts by inducing vasoconstriction of the meningeal arteries. It has been shown in pharmacological tests to act selectively as a potent agonist of serotonin 5-HT1B/1D receptors. Frovatriptan has been well tolerated in humans and efficacious in reducing headache pain and duration in clinical trials, which have also indicated that dose adjustments for age or gender are not necessary for the drug. Patients have found the use of frovatriptan acceptable over the long-term, and overall a low incidence of adverse effects has been reported. Though not a prophylactic, frovatriptan has demonstrated the potential to significantly improve the therapeutic approaches to the treatment of migraine. PMID- 12582450 TI - Therapeutic approaches to the treatment of Alzheimer's disease. AB - Alzheimer's disease is the most common cause of progressive decline of cognitive function in aged humans and is characterized by the presence of numerous senile plaques and neurofibrillary tangles accompanied by neuronal loss. The only treatment currently available for the disease is pharmacotherapy with acetylcholinesterase inhibitors, a palliative strategy aimed at the temporary improvement of cognitive function. Other strategies with disease-modifying potential may include the use of antiinflammatory drugs, estrogen replacement therapy and antioxidants. Recent progress in understanding the molecular and cellular pathophysiology of Alzheimer's disease has suggested possible pharmacological interventions that could modify the development and progress of the disease (disease-modifying therapy), such as treatment with secretase inhibitors, transition metal chelators, HMG-CoA reductase inhibitors and amyloid b immunization. Inhibitors of tau hyperphosphorylation may also modulate the development and progress of the disease. PMID- 12582451 TI - Reinventing clinical nuclear medicine. Highlights of the 49th annual meeting of the Society of Nuclear Medicine, Los Angeles, California, U.S.A. June 15-19, 2002. PMID- 12582453 TI - Identification of parkinsonism and Parkinson's disease. AB - Parkinson's disease is a common movement disorder associated with considerable disability. The clinical syndrome of parkinsonism is based on the presence of core clinical features of rest tremor, bradykinesia, rigidity and impaired postural reflexes or gait. Parkinsonism is most often caused by Parkinson's disease, but can also be caused by other disorders, including cerebrovascular disease, multiple-system atrophy, progressive supranuclear palsy and other disorders. Parkinsonism can be identified by questionnaires and confirmed in person or by videotaped clinical examinations. The identification of presymptomatic cases remains problematic but is motivated by the hope for treatment before symptoms appear. Quantitative approaches to the diagnosis of parkinsonism based on the measurement of the cardinal features are available. Clinical approaches should include identification of features atypical for Parkinson's disease, which exclude the diagnosis, and documentation of a response to dopaminergic medications, which support a diagnosis of Parkinson's disease. Loss of smell and visual dysfunction are found in early patients and may be useful in screening protocols. In addition, behavioral changes, including depressive symptoms, may be detected in presymptomatic cases. Cognitive changes, such as impaired set shifting, have been observed in early Parkinson's disease, but can be seen with other causes of parkinsonism. Neuroimaging techniques, including positron emission tomography or single-photon emission computed tomography, are available to quantify dopaminergic neurons, while magnetic resonance imaging may be helpful in differentiating other forms of parkinsonism from Parkinson's disease. There are numerous approaches available to the identification of parkinsonism and Parkinson's disease. The gold standard remains a clinical diagnosis, confirmed by autopsy. PMID- 12582452 TI - Newer GABAergic agents for pharmacotherapy of infantile spasms. AB - Infantile spasms is an age-specific epileptic syndrome in infants and young children. Although the exact mechanism is unknown, adrenocorticotrophic hormone (ACTH) has been the mainstay for the therapeutic management of infantile spasms and other developmental epilepsies. Clinical benefits of ACTH in infantile spasms could partially relate to its stimulatory effects on the release of adrenocorticosteroids and neurosteroids. Glucocorticoids, pyridoxine and ketogenic diet therapy have all been used for the treatment of refractory infantile spasms. Recent studies indicate that several newer anticonvulsant agents, which are positive allosteric modulators of GABA(A) receptors, are as effective as ACTH in acutely controlling infantile spasms. The efficacy of agents that enhance GABA-mediated inhibition (such as vigabatrin and benzodiazepines) for rapid and complete abolition of infantile spasms has been demonstrated in several clinical studies. Ganaxolone, a novel neuroactive steroid has, however, demonstrated outstanding efficacy and better tolerability in children with intractable infantile spasms. Zonisamide, topiramate, deoxycorticosterone and neurosteroids are emerging as effective treatment approaches. These new antiepileptic drugs represent a potential nonhormonal approach for infantile spasms, but additional studies are needed to verify their efficacy and tolerability. Future studies will hopefully identify rational antiseizure drugs that not only control infantile spasms but also abrogate its risk on the development of the brain. PMID- 12582454 TI - Postoperative chylous ascites--the urologist's view. AB - Postoperative chylous ascites is a rare complication of retroperitoneal surgery caused by incidental disruption of major retroperitoneal lymphatics and is associated with serious mechanical, nutritional and immunological consequences due to the constant loss of protein and lymphocytes. Different management schemes may be used successfully to treat chylous ascites. Treatment should be adjusted to the severity of the lymphatic leak and its consequences. The management algorithm presented in this article is based on a step-up approach aimed at reducing lymph flow and maintaining nutritional balance. It integrates repeated palliative paracenteses, dietary interventions, total parenteral nutrition therapy, somatostatin therapy, surgical closure of the lymphatic fistula and peritoneovenous shunting. The prognosis of chylous ascites is generally good, with the majority of the cases responding to conservative measures. PMID- 12582455 TI - The effects of cytokines on the atherosclerotic process. AB - Atherosclerotic lesions result from a series of highly specific cellular and molecular responses to various endogenous risk factors and potential exogenous antigens. The cellular mechanisms involved in atherogenesis, with the exception of calcification and thrombotic events, are principally no different to those found in chronic inflammatory fibroproliferative diseases such as liver cirrhosis, rheumatoid arthritis, glomerulosclerosis, pulmonary fibrosis or chronic pancreatitis. These responses are mediated by interactions among endothelial cells, monocyte-derived macrophages, smooth muscle cells and specific subtypes of T lymphocytes. Monocyte and lymphocyte activation leads to the release of a wide spectrum of cytokines and chemokines that have key roles in all of the phases of endothelial damage, as well as in the formation and rupture of the atherosclerotic plaques. This review attempts to analyze the role of chemokines and cytokines in the multiple steps of atherosclerotic process. PMID- 12582456 TI - Gene expression alterations in human prostate cancer. AB - Prostate cancer is a disease with a great degree of variation in biological aggressiveness and clinical prognosis. Although more than 30% of the older-aged male population develops prostate cancer, defined by histologic examination, a large number of these cases does not reach the stage displaying clinical symptoms. Among those patients with clinical prostate cancer, only a fraction of cases demonstrate life-threatening biological aggressiveness. Parallel to the clinical complexity of this disease, abnormalities in the prostate cancer genome have been reported in 21 of 23 pairs of human chromosomes, but none can be accountable for the dominant event in the development of prostate cancer. In order to understand the genetic nature of this disease, a comprehensive analysis of its gene expression patterns is needed. This article will review several recent publications in the area of gene expression analysis using microarray technology. I will discuss some of our findings in the area of gene expression alteration in benign prostate tissue adjacent to prostate cancer. The implication of these studies in potential clinical application will be explored. PMID- 12582457 TI - The therapeutic spectrum of infliximab and tumor necrosis factor immunomodulation in chronic inflammatory diseases. AB - This review summarizes the clinical results of Infliximab for the treatment of Crohn's disease, the limited data of its use in other digestive disorders and disorders related to the gastrointestinal tract. The etiology of these disorders remains enigmatic. As long as the precise etiology remains unknown, therapeutic options to treat these patients are predominantly symptomatic (i.e., immunosuppressive drugs or surgical intervention). To be able to develop more rational therapeutic regimens, it is pivotal to understand the immunopathological events that lead to, or are involved in, the pathogenesis of these diseases. Therefore, we also briefly summarize what at the present is known in this respect. PMID- 12582458 TI - Obesity and metabolic syndrome: long-term benefits of central leptin gene therapy. AB - The recent rapid rise in the incidence of obesity has prompted investigations into understanding the hormonal and neuronal pathways involved in body weight homeostasis in order to devise novel therapeutic strategies. The early enthusiasm for the adipocyte hormone leptin as a regulator of fat mass was largely discarded because of the apparent development of leptin resistance, as seen in obese subjects with elevated blood leptin levels. We postulated that this leptin ineffectiveness may be caused by a lack of leptin availability at target sites in the hypothalamus. To test this hypothesis, we used viral vectors to introduce the leptin gene into the brain for a sustained supply of leptin in the hypothalamus. A single injection of recombinant adeno-associated virus encoding the leptin gene (rAAV-lep) into the third cerebroventricle prevented the aging-associated gradual increase in body weight and adiposity in adult rats for 6 months of the experiment. When administered to prepubertal rats, significantly lower body weight gain and adiposity were maintained for up to 10 months of the experiment. In addition, obesity was prevented in rats introduced to a high-fat diet and also reversed in obese-prone rats maintained on a high-fat diet. Body weight homeostasis and loss of adiposity by leptin gene therapy was achieved by an increase in energy expenditure, and when the rAAV-lep titer was increased, there was also a voluntary reduction in food intake. Importantly, this therapy reduced blood levels of insulin, triglycerides and free fatty acids, the pathophysiologic correlates of the metabolic syndrome. Thus, the long-term beneficial effects of central leptin gene therapy may herald the development of newer therapeutic strategies to control the epidemic of obesity and related metabolic disorders. PMID- 12582459 TI - The use of carbohydrate antigens for the preparation of vaccines for therapy in breast cancer. AB - Immunotherapy of human breast cancer is a rapidly growing experimental area based on peptidic as well as carbohydrate tumor-associated antigens. Cell surface carbohydrates are characteristic of different stages of normal development and differentiation; distinct carbohydrates are expressed in tissue- and cell specific manners during those processes. Under pathological conditions such as neoplasia, changes in carbohydrate structures can almost always be present. In the successful development of carbohydrate anticancer vaccines, adjuvant and carrier molecules that promote the presentation of an antigen to the immune effector cells are prominent factors. At present, keyhole limpet hemocyanin (KLH) is the most widely used carrier protein in cancer immunotherapy, and QS-21 is the adjuvant most widely employed. In breast cancer, the immunogenicity of TF, Tn, STn and globo H antigens has been explored in different clinical trials. Approaches including vaccines against STn and globo H are presently being assayed with expectancy. From the experience obtained, it is possible to conclude that: 1) the employment of glycolipopeptide with clustered epitopes seems to be more effective than that of related structures with single epitopes in inducing antitumor cell antibodies; 2) the conjugation to carriers is best achieved using KLH; 3) totally synthetic constructs can be better immunogens in conjunction with a suitable adjuvant such as QS-21; in some cases, this adjuvant leads to a bypass in the need for specific T-cell help to stimulate IgG as well as IgM antibodies. In other cases, a T-cell mediated immune response is obtained, and (4) the development of a totally synthetic vaccine would greatly facilitate the production of the vaccine for large scale clinical trials with the attainment of regulatory approval. PMID- 12582460 TI - The impact of diabetes and prediabetes on risk of cardiovascular disease and mortality. AB - Although it is well established that type 2 diabetes is a strong risk factor for coronary heart disease, it is still a matter of debate whether diabetes and a prior coronary heart disease confer similar risk for future risk of fatal coronary heart disease. Most of the published studies on this topic did not consider the duration of diabetes when comparing the effects of diabetes alone and a prior coronary heart disease alone. Recent studies suggest that duration of clinical diabetes is an important risk factor for fatal coronary heart disease and total mortality. In addition, the incidence of cardiovascular disease is elevated long before the clinical diagnosis of diabetes. PMID- 12582461 TI - An unexpected association between urinary incontinence, depression and sexual dysfunction. AB - Certain aspects of urinary incontinence, depression and sexual function are proving to be tied to one another in ways other than the obvious. From a biochemical standpoint certain monoamines, specifically serotonin, appear to be important in the expression of these pathologic states. In depression, the role of serotonin is fairly well established, and there is increasing evidence for there being a role in a specific cause of incontinence as well. There is only empiric evidence that serotonin plays a role in some types of sexual dysfunction. However, the possibility of this connection between such distinct disorders is fodder for theoretic conjecture. PMID- 12582462 TI - Somatostatin analogues as drug therapies for retinopathies. AB - Proliferative retinopathies account for the majority of cases of vision loss throughout the world. Currently accepted therapy for retinopathy consists of retinal ablation by panretinal laser photocoagulation or cryotherapy. This technique is not without deleterious effects to patients, including diminished night vision, reduced peripheral vision and loss of precise vision, decreasing visual acuity by one to two lines in magnitude. One promising area of research into pharmacotherapeutics for retinopathies, especially proliferative diabetic retinopathy, involves the use of synthetic analogues of somatostatin. The rationale for somatostatin as a therapeutic agent for retinal neovascularization is discussed. Somatostatin analogues such as octreotide have shown promise as a safe and effective treatment for severe proliferative diabetic retinopathy by blocking the local and systemic production of growth hormone and insulin-like growth factor type 1 associated with angiogenesis and endothelial cell proliferation. There are also observations suggesting an autocrine and paracrine effect of somatostatin, perhaps directly on retinal cells, which are known to express somatostatin receptors (SSTR). SSTR2 and SSTR3 are the most important receptor subtypes mediating growth hormone secretion and endothelial cell cycle arrest, retinal endothelial cell apoptosis and release of insulin. Thus, analogues that target these receptor subtypes may prove more useful. Long-acting somatostatin analogues are currently being tested for treatment of diabetic retinopathy and are, in fact, the only therapeutic alternative for patients who fail panretinal photocoagulation. Whether such a therapy may also prove effective for other retinal vascular proliferative diseases such as retinopathy of prematurity and age-related macular degeneration remains an open question that deserves attention, given our new understanding of the cellular and molecular mechanisms by which somatostatin may exert its antiangiogenic effects. PMID- 12582463 TI - The potential of gene therapy in the treatment of pancreatic cancer. AB - Pancreatic cancer is known to be one of the most problematic forms of cancer. Researchers around the world are still trying to find an adequate treatment for this disease. While surgical operation has been the dominant procedure for treating pancreatic cancer, adjuvant therapies such as radiation or chemotherapy (although the survival rate is still poor) also exist. Pancreatic cancer is notoriously difficult to detect at its initial invasion, despite modern radiographic technology. This means that patients discover the cancer when it is already in an advanced stage, making surgical resection difficult. A new strategy for medical practice in pancreatic cancer is much needed. Gene therapy is currently in the spotlight as a promising new method for cancer cure. Many studies have revealed the potential of this therapy for the treatment of pancreatic cancer, and early clinical trials are taking place to evaluate the success of gene therapy regimes in humans. Here we discuss basic scientific principles and clinical experience with respect to these regimes, including antisense strategies, gene- directed prodrug activation therapy, promoter-gene strategies and oncolytic viral therapy. PMID- 12582464 TI - The clinical potentials of endogenous neurosteroids. AB - Stress increases plasma and brain concentrations of the neurosteroids allopregnanolone and allotetrahydrodeoxycorticosterone (THDOC), which can have potent effects on GABAA receptors in the brain. Blockade of the formation of neurosteroids prevents specific biochemical and behavioral effects of stress, suggesting that those effects are dependent upon the actions of GABA(A)-receptor active neurosteroids. Recent investigations provide a better understanding of the role of endogenous neurosteroids in normal neuronal development and in the pathophysiology of brain disorders. Physiological neurosteroid fluctuations have potential implications for stress-sensitive neurological conditions such as epilepsy, infantile spasms, as well as psychiatric disorders such as schizophrenia, posttraumatic stress disorder and depression. Future studies may provide important new evidence that may not only explain acute actions of stress, but also reveal the clinical importance of neurosteroid mechanisms during chronic stress. PMID- 12582465 TI - Female urinary incontinence: diagnosis, treatment and patients' concerns. AB - We developed a simple and accurate method of diagnosing urinary incontinence by means of a 15-item questionnaire that helps physicians evaluate a patient's incontinence status more easily than conventional diagnostic procedures. The results of using the questionnaire showed a total improvement rate of more than 80% among urinary incontinence patients treated in our clinic. Surgical treatment for stress urinary incontinence has improved in terms of both effectiveness and patient quality of life after introduction of a tension-free vaginal tape procedure. Our research on the prevalence of urinary incontinence among a large number of Japanese women showed that, overall, 26.8% complained of some symptoms of incontinence and, the percentage increased with age, reaching more than 40% among women over 40 years of age. However, very few of the women (4%) wished to consult a physician, with shame in the young population and resignation in the old population being the major reasons for not seeking treatment. Women need to be provided with correct information about urinary incontinence in order to improve their quality of life. PMID- 12582466 TI - Recent developments in antimutagenesis and anticarcinogenesis. PMID- 12582467 TI - Nonplatelet-mediated effects of aspirin. AB - Aspirin has nonplatelet-mediated effects that contribute to its efficacy in the primary and secondary prevention of coronary events. These include antiarrhythmic effects, as shown in animal studies, and antiatherosclerotic effects related to increase in nitric oxide synthesis/activity and reduction in inflammatory mediators. Epidemiological studies have also shown primary antiinflammatory properties. Aspirin is known to inhibit vascular smooth muscle cell proliferation and to produce an endothelial stabilizing effect. Other observed outcomes from the administration of this compound include a modest anticoagulant activity, angiogenesis reduction and a decrease in oxidant stress. We believe that these results complement the antiplatelet effect and make this agent unique in the management of ischemic heart disease. PMID- 12582468 TI - Gene expression in Alzheimer's disease. AB - Even though the exact cause of Alzheimer's disease is not known, it is clear that genetic factors play a major role. Screening of familial cases has so far linked four genes to Alzheimer's disease: amyloid precursor protein, presenilin 1, presenilin 2 and apolipoprotein E. Mutations in the amyloid precursor protein, presenilin 1 and presenilin 2 genes cause the enhanced production of b amyloid that is found in neuritic plaques. While apolipoprotein E allele e4 does not cause enhanced production of amyloid, it does enhance its deposition. The genes identified so far are linked to only about 10% of total Alzheimer's disease cases, and there are a number of familial cases that are not linked to any of the four genes. This suggests that important genetic factors have yet to be identified. The key events in Alzheimer's disease are cytoskeletal changes and the formation of paired helical filaments, and these are common to the two hallmarks of Alzheimer's disease: neuritic plaques and neurofibrillary tangles. So far there is no explanation as to how the identified genes cause these events. Here we speculate that paired helical filaments can form as a result of overexpression of the DSCR1 (Adap78) gene. b Amyloids and/or other stress factors can induce DSCR1 (Adap78) and cause decreased activity of calcineurin. Chronic downregulation of calcineurin can lead to gradual accumulation of hyperphosphorylated tau, the formation of paired helical filaments and Alzheimer's disease-like cytoskeletal changes. PMID- 12582469 TI - Recent advances in nephrology: highlights from the 35th annual meeting of the American society of nephrology. AB - The 35th Annual Meeting of the American Society of Nephrology, held in Philadelphia, Pennsylvania, United States (October 30 to November 4, 2002) presented the newest advances in basic and clinical nephrology science. Several presentations and symposia discussed the effects of various interventions and risk factors in clinical outcomes in dialysis patients. The recent evidences of pure red cell aplasia secondary to neutralizing antibodies against erythropoietin were also extensively discussed in a special symposium. Recent advances in the management of calcium phosphorus metabolism and secondary hyperparathyroidism, such as the clinical efficacy and safety of AMG-073, a new calcimimetic agent in the control of hyperparathyroidism in chronic kidney disease patients, or the use of sevelamer or lanthanum carbonate as phosphate binders, were presented. The results in animal models on improved sparing of renal function with rapamycin versus cyclosporin A represent a promising advance in renal transplantation. Finally, the recent discoveries with the newly identified disease gene PKHD1, which causes autosomal recessive polycystic kidney disease, were also presented at the meeting. PMID- 12582471 TI - Angiogenic gene therapy. AB - Angiogenesis, which is defined as the outgrowth of nutritive vessels from locally preexisting ones, is primarily based on the outgrowth of locally present endothelial cells and implies a delicate balance where both stimulating and inhibitory factors can influence the outcome. The primary players starting the process are the vascular endothelial growth factors (VEGFs), which stimulate endothelial cell growth, although some other key factors such as fibroblast growth factors (FGFs), placental growth factors (PIGFs), platelet-derived growth factors (PDGFs) and angiopoietins must come into play for vessels to mature and not deteriorate. Two possibilities exist in terms of local delivery of the angiogenic proteins. One is the delivery of the protein in a slow-release formulation. Fibrin or alginate formulations have been implanted, sub epicardially releasing FGF. Another approach to get a local delivery over a period of time is gene therapy by local delivery of the vector carrying the therapeutic gene or with the development of vectors that are taken up and expressed only in the organ of interest. A transient overexpression, desirable for the treatment of ischemic heart disease, can be achieved with adenoviral vectors or naked plasmid. With VEGF gene transfection, angiogenesis and reendothelialization were demonstrated in animal cardiac ischemia models showing proof of principle for cardiac therapy and paving the way for clinical trials. Currently, about 200 patients have been treated with intramyocardial VEGF gene therapy for peripheral occlusive artery disease or for myocardial ischemia. Reported adverse events have been few and no worsening of atherosclerosis following treatment has been observed. PMID- 12582470 TI - Leptin and the central neural mechanisms of obesity hypertension. AB - The prevalence of obesity is rising at an alarming rate worldwide, with consequent increases in type 2 diabetes, hypertension and cardiovascular morbidity and mortality. Central neural mechanisms, via the activation of the sympathetic nervous system may contribute to obesity-related cardiovascular diseases through the promotion of hypertension, dysrhythmia and atherosclerosis. However, the mechanisms responsible for this sympatho activation have not been identified. Leptin is an adipocyte-derived hormone that promotes weight loss by reducing appetite and by increasing energy expenditure through sympathetic stimulation to thermogenic tissue. Leptin also produces sympathoactivation to kidneys, hindlimb and adrenal glands, suggesting that the obesity-associated increase in sympathetic nerve activity could be due in part to these sympathetic effects of leptin. However, most human obesity appears to be associated with leptin resistance. Recent studies indicate that leptin resistance may be selective, with preservation of adverse sympathetic effects despite the loss of the metabolic actions of leptin. The leptin receptor is expressed in several hypothalamic nuclei including the arcuate nucleus. The melanocortin system, neuropeptide Y and corticotrophin-releasing factor have emerged as principal neuropeptide mediators of leptin action in the arcuate nucleus. These neuropeptides exert varying effects by different pathways. Several other candidate hypothalamic pathways that can mediate the effects of leptin have been identified. The understanding of neuronal signaling pathways involved in leptin signaling and energy balance has opened new research possibilities for the treatment of obesity. PMID- 12582472 TI - Caspofungin. AB - Available systemically effective antifungal agents for the treatment of invasive fungal infections are few. With the increasing recognition of a need for newer antifungal drugs, caspofungin has been introduced as the first member of a new class of compounds called echinocandins. This paper reviews the chemistry and mechanism of action of caspofungin, its activity in vitro and in animal models, and clinical pharmacokinetics,clinical efficacy and safety in patients. PMID- 12582473 TI - Medical backgrounder: psoriasis. AB - Psoriasis is a chronic skin disorder that affects approximately 2% of the US and European populations. Psoriatic lesions are extremely characteristic of the disease, which allows for simple diagnosis. A clear understanding of the pathogenesis of psoriasis does not yet exist. Hyperproliferation of keratinocytes is a further characteristic feature. Studies have depicted that the epidermal cell cycle of psoriatic lesions is shortened by approximately eight-fold more than normal. The lesions are classified as erythrosquamous, due to the erythema which develops asa result of involvement of the vasculature, and the involvement of the epidermis with scale formation. The diagnosis of psoriasis can usually be established on clinical grounds. If the clinician is in doubt, a small cutaneous punch biopsy and subsequent histopathological examination can be performed. There are multiple therapeutic options for psoriasis. First-line therapy for patients with moderate to severe psoriasis is the application of topical agents, followed by phototherapy (UVB) for more extensive disease. If extensive disease does not respond to UVB, second-line agents include psoralen plus UVA (PUVA), methotrexate, cyclosporine or other systemic agents, including novel biologic therapies. New psoriasis treatment regimens have been developed and include combination, rotational and sequential therapy. PMID- 12582474 TI - The pathophysiology of benign prostatic hyperplasia. AB - The development of human benign prostatic hyperplasia and its related signs and symptoms clearly requires a combination of testicular androgens and aging. Although the role of androgens as a causative factor for human benign prostatic hyperplasia is debated, they undoubtedly play at least a permissive role. The principle prostatic androgen is dihydrotestosterone. Two isoenzymes of 5-alpha reductase have been discovered. Type 2 is dominant in the genital issue. Testosterone is reduced by the 5-a reductase to dihydrotestosterone. Benign prostatic hyperplasia is predominantly due to stromal hyperplasia of the gland and affects more than 70% of men of 70 years or older with or without obstruction. Recent studies identified transforming growth factor-b, fibroblast growth factor and insulin-like growth factor family members as key regulators of cell proliferation and extracellular matrix turnover with interrelated activities. Furthermore, estrogens, andenergic signaling and inflammatory processes have been shown to have an impact. Without a solid understanding of the physiology and pathophysiology of benign prostatic hyperplasia it can be difficult to interpret the results of clinical trials and symptoms. PMID- 12582476 TI - Diversity of chloroplast DNA SSRs in wild and cultivated soybeans: evidence for multiple origins of cultivated soybean. AB - Soybean [ Glycine max (L.) Merr.] is one of the major crops in the world and was domesticated from a wild progenitor, Glycine soja Sieb. & Zucc., in East Asia. In order to address the questions concerning the evolution and maternal lineage of soybean, we surveyed the variation in chloroplast DNA simple sequence repeats (cpSSR) of 326 wild and cultivated soybean accessions that were collected from various Asian countries. Twenty-three variants were detected at six cpSSRs in the accessions tested. All of the variants were found in wild soybean, whereas only 14 variants existed in the cultigen. Combining the variants at the six cpSSRs gave 52 haplotypes in the former and eight haplotypes in the latter. Both analyses indicated a considerably higher genetic diversity in the wild soybean. Around 75% of the cultivated accessions tested possessed a common haplotype (no. 49), which was detected in only seven wild accessions, six from southern Japan and one from southern China. The predominant haplotype in the cultigen may therefore have originated from a rare haplotype of the wild soybean that is presently distributed in the southern areas of Japan and China. The remaining seven haplotypes in the cultigen were distributed regionally, and except for three rare haplotypes, largely overlapped with the distributions of wild accessions with the same respective haplotypes. Our results strongly suggest that the cultivated soybeans with different cpDNA haplotypes originated independently in different regions from different wild gene pools and/or hybrid swarms between cultivated and wild forms. PMID- 12582477 TI - A genetic and physical map of the region containing PLASTOCHRON1, a heterochronic gene, in rice ( Oryza sativa L.). AB - The rice heterochronic gene plastochron1, pla1, shows shorter plastochron and ectopic expression of the vegetative program during the rice reproductive phase resulting in aberrant panicle formation. A genetic and physical map was constructed to isolate the causal gene for the pla1 syndrome. Small-scale mapping was carried out to determine the approximate map position of the pla1 locus, and then a high-resolution genetic map was made for pla1-1, one of the pla1 alleles, using an F(2) population comprising 578 pla1-1 homozygous plants. In a high resolution genetic map, the pla1-1 locus was found to map between RFLP markers C961 and R1738A on chromosome 10, within a 3.6-cM genetic distance. A physical map encompassing the pla1-1 locus was constructed by overlapping Bacterial Artificial Chromosome (BAC) clones through chromosome walking. PCR-based RFLP markers from BAC-end clones were developed and mapped relative to the pla1 locus. Physical map construction using BAC clones indicated that a BAC clone, B44A10 (167-kb), contained the pla1 locus within 74-kb corresponding to a 0.52-cM genetic distance. Gene prediction of 74-kb region carrying the pla1 locus suggested several candidate genes for the pla1 gene. Identification of a candidate gene for pla1 will be made by sequence analysis of allele variation and cDNA screening. PMID- 12582478 TI - Integration of microsatellite markers into the translocation-based physical RFLP map of barley chromosome 3H. AB - PCR with the DNA of translocation chromosomes and marker-specific primers has been used to merge genetically mapped microsatellite (MS) markers into the physically integrated restriction fragment length polymorphism (RFLP) map of barley chromosome 3H. It was shown that the pronounced clustering of MS markers around the centromeric region within the genetic map of this chromosome results from suppressed recombination. This yielded a refinement of the physically integrated RFLP map of chromosome 3H by subdivision of translocation breakpoints (TBs) that were previously not separated by markers. The physical distribution of MS markers within most of the subchromosomal regions corresponded well with that of the RFLP markers, indicating that both types of markers are similarly valuable for a wide range of applications in barley genetics. PMID- 12582479 TI - Assessment of genetic diversity within and between pearl millet landraces. AB - A minimum core subset of pearl millet [ Pennisetum glaucum (L.) R. Br.], which comprised 504 landrace accessions, was recently established from the global pearl millet germplasm collection of ICRISAT. The accessions for this core were selected by a random proportional sampling strategy following stratification of the entire landrace collection (about 16,000 accessions) according to their geographic origin and morpho-agronomic traits. In this study RFLP probes were used to quantify the genetic diversity within and between landrace accessions of this minimum core using a subset comprising ten accessions of Indian origin. Twenty five plants per accession were assayed with EcoRI, EcoRV, HindIII and DraI restriction enzymes, and 16 highly polymorphic RFLP probes, nine associated with a quantitative trait loci (QTLs) for downy mildew resistance, and five associated with a QTL for drought tolerance. A total of 51 alleles were detected using 16 different probe-enzyme combinations. The partitioning of variance components based on the analysis of molecular variance (AMOVA) for diversity analysis revealed high within-accession variability (30.9%), but the variability between accessions was significantly higher (69.1%) than that within the accessions. A dendrogram based on the dissimilarity matrix obtained using Ward's algorithm further delineated the 250 plants into ten major clusters, each comprised of plants from a single accession (with the exception of two single plants). A similar result was found in an earlier study using morpho-agronomic traits and geographic origin. This study demonstrated the utility of RFLP markers in detecting polymorphism and estimating genetic diversity in a highly cross pollinated species such as pearl millet. When less-tedious marker systems are available, this method could be further extended to assess the genetic diversity between and within the remaining accessions in the pearl millet core subset. PMID- 12582480 TI - Transmission of herbicide resistance from a monoecious to a dioecious weedy Amaranthus species. AB - The genus Amaranthus includes several important monoecious and dioecious weed species, and several populations of these species have developed resistance to herbicides. These species are closely related and two or more species often coexist in agricultural settings. Collectively, these attributes raise the concern that herbicide resistance might transfer from one weedy Amaranthus species to another. We performed research to determine if a dominant allele encoding a herbicide-insensitive form of acetolactate synthase (ALS) could be transferred from a monoecious species, A. hybridus, to a dioecious species, A. rudis. Numerous F(1) hybrids were obtained from controlled crosses in a greenhouse between A. rudis and herbicide-resistant A. hybridus, and most (85%) of these hybrids were herbicide-resistant. Molecular analysis of the ALS gene was used to verify that herbicide-resistant hybrids contained both an A. rudis and an A. hybridus ALS allele. Although hybrids had greatly reduced fertility, 42 BC(1) plants were obtained by backcrossing 33 hybrids with male A. rudis. Fertility was greatly restored in BC(1) progeny, and numerous BC(2) progeny were obtained from a second backcross to A. rudis. The herbicide-resistance allele from A. hybridus was transmitted to 50% of the BC(1) progeny. The resistance allele was subsequently transmitted to and conferred herbicide resistance in 39 of 110 plants analyzed from four BC(2) families. Parental species, hybrids, and BC(2) progeny were compared for 2C nuclear DNA contents. The mean hybrid 2C nuclear DNA content, 1.27 pg, was equal to the average between A. rudis and A. hybridus, which had 2C DNA contents of 1.42 and 1.12 pg, respectively. The mean 2C DNA content of BC(2) plants, 1.40 pg, was significantly (alpha < 0.01) less than that of the recurring A. rudis parent and indicated that BC(2) plants were not polyploid. This report demonstrates that herbicide resistance can be acquired by A. rudis through a hybridization event with A. hybridus. PMID- 12582481 TI - Molecular characterization of some Indian Basmati and other elite rice genotypes using fluorescent-AFLP. AB - Cultivated rice is a high-volume, low-value cereal crop providing staple food to more than 50% of the world populace. A small group of rice cultivars, traditionally produced on the Indo-Gangetic plains and popularly known as Basmati, have exquisite quality grain characteristics and are a prized commercial commodity. Efforts to improve the yield potential of Basmati have led to the development of several crossbred Basmati-like cultivars. In this study we have analysed the genetic diversity and interrelationships among 33 rice genotypes consisting of the traditional Basmati, improved Basmati-like genotypes developed in India and elsewhere, American long-grain rice and a few non-aromatic rice using a DNA marker-based approach - fluorescent-amplified fragment length polymorphism (f-AFLP). Using a set of nine primer-pairs we scored a total of 10,672 data points over all of the genotypes in the size range of 75-500 bp. The scored data points corresponded to a total of 501 AFLP markers (putative loci/genome landmarks) of which 327 markers (65%) were polymorphic. The f-AFLP marker data, which were analysed using different clustering algorithms and principal component analysis, indicate that: (1) considerable genetic variability exists in the analysed genotypes; (2) traditional Basmati cultivars could be distinctly separated from the crossbred Basmati-like genotypes as well as from the non-aromatic rice; (3) the crossbred Basmati-like cultivars from the subcontinent and elsewhere are genetically very distinct; (4) f-AFLP-based clustering, in general, conforms to the putative pedigree of the improved genotypes. Moreover, analysis to ascertain the scope of AFLP as a technique suggests that the polymorphism revealed by three selective primer-pair combinations is sufficient to obtain reliable estimates of genetic diversity for the type of material used in this study. However, its utility to identify group specific DNA markers was discounted due to a low frequency of observed group specific discrete markers. PMID- 12582482 TI - Identification and mapping of an AFLP marker linked to Gm7, a gall midge resistance gene and its conversion to a SCAR marker for its utility in marker aided selection in rice. AB - We have identified an AFLP marker SA598 that is linked to Gm7, a gene conferring resistance to biotypes 1, 2 and 4 of the gall midge ( Orseolia oryzae), a major dipteran pest of rice. A set of PCR primers specific to an RFLP marker, previously identified to be linked to another gall midge resistance gene Gm2, also amplified a 1.5-kb (F8LB) fragment that is linked to Gm7. Gm7 is a dominant gene and non-allelic to Gm2. Hybridization experiments with clones from a YAC library of Nipponbare, a japonica variety, a BAC library of IR-BB21, an indica variety, and cosmid clones encompassing Gm2 from Phalguna, an indica variety, with F8LB and SA598 as probes, revealed that Gm7 is tightly linked to Gm2 and is located on chromosome 4 of rice. SA598 was sequenced and the sequence information was used to design sequence-characterized amplified region (SCAR) primers. The potential use of these SCAR primers in marker-aided selection of Gm7 in a rice breeding program has been demonstrated. PMID- 12582483 TI - Assessing genetic diversity of wheat ( Triticum aestivum L.) germplasm using microsatellite markers. AB - A set of 24 wheat microsatellite markers, representing at least one marker from each chromosome, was used for the assessment of genetic diversity in 998 accessions of hexaploid bread wheat ( Triticum aestivum L.) which originated from 68 countries of five continents. A total of 470 alleles were detected with an average allele number of 18.1 per locus. The highest number of alleles per locus was detected in the B genome with 19.9, compared to 17.4 and 16.5 for genomes A and D, respectively. The lowest allele number per locus among the seven homoeologous groups was observed in group 4. Greater genetic variation exists in the non-centromeric regions than in the centromeric regions of chromosomes. Allele numbers increased with the repeat number of the microsatellites used and their relative distance from the centromere, and was not dependent on the motif of microsatellites. Gene diversity was correlated with the number of alleles. Gene diversity according to Nei for the 26 microsatellite loci varied from 0.43 to 0.94 with an average of 0.77, and was 0.78, 0.81 and 0.73 for three genomes A, B and D, respectively. Alleles for each locus were present in regular two or three base-pair steps, indicating that the genetic variation during the wheat evolution occurred step by step in a continuous manner. In most cases, allele frequencies showed a normal distribution. Comparative analysis of microsatellite diversity among the eight geographical regions revealed that the accessions from the Near East and the Middle East exhibited more genetic diversity than those from the other regions. Greater diversity was found in Southeast Europe than in North and Southwest Europe. The present study also indicates that microsatellite markers permit the fast and high throughput fingerprinting of large numbers of accessions from a germplasm collection in order to assess genetic diversity. PMID- 12582484 TI - Identification of QTLs controlling tissue-culture traits in barley ( Hordeum vulgare L.). AB - Quantitative trait loci (QTLs) controlling callus growth (CG), subsequent shoot differentiation ratio (SD) and green shoot ratio (GS) in immature embryo culture were identified in barley. A base map was developed from 99 recombinant inbred lines (RILs) of 'Azumamugi' x 'Kanto Nakate Gold'. The tissue-culture traits were evaluated at the F(7) and F(10) generations of the RILs. The RILs showed wide and continuous variations in each of the three tissue-culture traits. Three QTLs for CG, three QTLs for SD and two QTLs for GS were detected by using composite interval mapping. A QTL for SD on chromosome 3H had a large effect, and 'Kanto Nakate Gold', which has a high differentiation ability, contributed to this QTL. The location of this QTL is identical to, or very close to, the uzu locus. We discuss the relationships between tissue-culture loci in 'Azumamugi' x 'Kanto Nakate Gold' and those in other mapping populations. PMID- 12582485 TI - A genetic linkage map of Guinea yam ( Dioscorea rotundata Poir.) based on AFLP markers. AB - A genetic linkage map of the tetraploid white yam ( Dioscorea rotundata Poir.) was constructed based on 341 co-dominantly scored amplified fragment length polymorphism (AFLP) markers segregating in an intraspecific F(1) cross. The F(1) mapping population was produced by crossing a landrace cultivar TDr 93-1 as female parent to a breeding line TDr 87/00211 as the male parent. The marker segregation data were split into maternal and paternal data sets, and separate genetic linkage maps were constructed since the mapping population was an F(1) cross between two presumed heterozygous parents. The markers segregated like a diploid cross-pollinator population suggesting that the D. rotundata genome is an allo-tetraploid (2n = 4 x = 40). The maternal map comprised 155 markers mapped on 12 linkage groups with a total map length of 891 cM. Three linkage groups consisted of maternal parent markers only. The paternal map consisted of 157 markers mapped on 13 linkage groups with a total map length of 852 cM. Three and one quantitative trait loci (QTLs) with effects on resistance to Yam Mosaic Virus (YMV) were identified on the maternal and paternal linkage maps, respectively. Prospects for detecting more QTLs and using marker-assisted selection in white yam breeding appear good, but this is subject to the identification of additional molecular markers to cover more of the genome. PMID- 12582486 TI - A genetic linkage map of water yam ( Dioscorea alata L.) based on AFLP markers and QTL analysis for anthracnose resistance. AB - A genetic linkage map of the tetraploid water yam ( Dioscorea alata L.) genome was constructed based on 469 co-dominantly scored amplified fragment length polymorphism (AFLP) markers segregating in an intraspecific F(1) cross. The F(1) was obtained by crossing two improved breeding lines, TDa 95/00328 as female parent and TDa 87/01091 as male parent. Since the mapping population was an F(1) cross between presumed heterozygous parents, marker segregation data from both parents were initially split into maternal and paternal data sets, and separate genetic linkage maps were constructed. Later, data analysis showed that this was not necessary and thus the combined markers from both parents were used to construct a genetic linkage map. The 469 markers were mapped on 20 linkage groups with a total map length of 1,233 cM and a mean marker spacing of 2.62 cM. The markers segregated like a diploid cross-pollinator population suggesting that the water yam genome is allo-tetraploid (2n = 4 x = 40). QTL mapping revealed one AFLP marker E-14/M52-307 located on linkage group 2 that was associated with anthracnose resistance, explaining 10% of the total phenotypic variance. This map covers 65% of the yam genome and is the first linkage map reported for D. alata. The map provides a tool for further genetic analysis of traits of agronomic importance and for using marker-assisted selection in D. alata breeding programmes. QTL mapping opens new avenues for accumulating anthracnose resistance genes in preferred D. alata cultivars. PMID- 12582487 TI - Studies of the transferability of microsatellites derived from Triticum tauschii to hexaploid wheat and to diploid related species using amplification, hybridization and sequence comparisons. AB - Hexaploid wheat ( Triticum aestivum L em Thell) is derived from a complex hybridization procedure involving three diploid species carrying the A, B and D genomes, respectively. We recently isolated microsatellites from a T. tauschii library enriched for various motifs and evaluated the transferability of these markers to several diploid species carrying the A, B or D genomes. All of the primer pairs amplifying more than one locus on bread wheat and half of those giving D-genome-specific loci gave an amplification product on A-and/or B-diploid species. All of the markers giving a single amplification product for T. tauschii and no amplification on the other diploid species were D-genome-specific at the hexaploid level. The non-specific microsatellite markers (which gave an amplification product on diploid species carrying the A, B or D genome) gave either a complex amplification pattern on bread wheat (with several bands) or generated a single band which mapped to the D genome. Southern blot hybridizations with probes corresponding to the microsatellite flanking regions gave a signal on all diploid and hexaploid species, whatever the specificity of the microsatellite. The patterns observed on bread wheat were generally in accordance with those observed for diploid species, with slight rearrangements. This suggests that the specificity of microsatellite markers is probably due to mutations in microsatellite flanking regions rather than sequence elimination during polyploidization events and that genome stringency is higher at the polyploid than at the diploid level. PMID- 12582488 TI - Molecular dissection of seedling-vigor and associated physiological traits in rice. AB - seedling-vigor is important for crop establishment. There have been reported quantitative trait locus (QTL) analyses on seedling-vigor related morphological traits. However, physiological understanding of these detected QTLs is rather limited. In this study, we employed a recombinant inbred population to detect QTLs for seedling-vigor traits and physiological traits related to seedling vigor. Germination rate and seedling growth were measured to quantify seedling vigor. Total amylase activity, alpha-amylase activity, reducing sugar content, root activity and seed weight were determined. Correlations were observed between the seedling-vigor and physiological traits. QTL analysis reveals that the intervals of RG393-C1087-RZ403 on chromosome 3, C246-RM26-C1447 and R830-R3166 RG360-C734b on chromosome 5, and the interval of Waxy on chromosome 6 are the four main chromosomal regions controlling seedling-vigor. Several QTLs for amylase activities, reducing sugar content and root activity were localized in the similar regions as the QTLs for seedling-vigor. The results suggest that these traits were under the control of pleiotropic and/or closely linked QTLs. The implications of the results in the understanding of the physiological basis of seedling-vigor were discussed. PMID- 12582489 TI - Intergeneric hybridization between Erucastrum canariense and Brassica rapa. Genetic relatedness between E(C) and A genomes. AB - An intergeneric hybrid between a wild species, Erucastrum canariense (2n = 18; E(C)E(C)), and a cultivated oilseed brassica species, Brassica rapa (2n = 20; AA), was synthesized through ovary culture in White's basal medium supplemented with casein hydrolysate. Morphological, cytological and DNA-based analysis helped to establish the hybrid nature of the derived plants. Hybrid plants were morphologically intermediate between the two parents and were completely male, as well as female sterile. Cytological analysis revealed the occurrence of 19 I in about 38% of the PMCs investigated. However 1-8 bivalents/PMC were also observed, indicating a significant level of homology between the two genomes. Normal chromosome pairing and pollen fertility was restored following colchiploidy. The intergeneric amphiploid developed during the investigation can be used as a bridging species for the transfer of desirable genes from E(C) to cultivated genomes (especially A and C), and for resistance to Alternaria blight and mustard aphid. Under field conditions, the E. canariense intergeneric hybrid and the amphiploid appeared to be moderately resistant to Alternaria blight and also harboured a significantly lower population of mustard aphid than the cultivated B. rapa. PMID- 12582490 TI - Comparative mapping of the wheat 5B short chromosome arm distal region with rice, relative to a crossability locus. AB - Colinearity between wheat and rice genomes is quite well established at the chromosome level, but less is known at a finer level. We tried to specify these relationships for the wheat 5BS chromosome-arm distal region, where a major locus for crossability was located. By developing AFLP markers, we succeeded to locate this major QTL more precisely. One cloned AFLP fragment mapped to rice chromosome 11, which was in agreement with a rice chromosome-11 linkage block reported in this region. However a second marker, a RFLP probe, showed a break in synteny because it mapped to rice long-arm chromosomes 1 and 5, while screening a rice BAC library with the same probe identified rice chromosomes 5 and 6. Therefore, we concluded that the syntenic relationships were more complex at the fine level. The observed results might indicate the presence of a linkage block carrying a crossability gene on wheat groups 1, 5 and 7, and also on rice chromosomes 5 and 6. PMID- 12582491 TI - Development of STS markers linked to Hessian fly resistance gene H6 in wheat. AB - Hessian fly is one of the world's most destructive insect pests of wheat Triticum aestivum L. We have used the combination of near-isogenic lines (NIL) and random amplified polymorphic DNA (RAPD) analysis to screen up to 2,000 primers to identify DNA markers that are linked to gene H6 that confers resistance to biotype B of the insect. This screen produced six primers that show polymorphic fragments associated with resistance by H6. We have screened 440 F(2) individuals from a cross of the susceptible cultivar Newton and a NIL that contains H6 to verify the linkage between these markers and the resistance gene. A high resolution genetic map was constructed based on recombination frequency. Two of the markers were tightly linked to the gene with no recombination observed, three were within 2.0 cM, and one was 11 cM from the gene. Three of the six markers were successfully converted to sequence tagged site (STS) markers. Both RAPD and STS primers were used to screen for the presence or absence of the resistance gene in wheat varieties. The identification of markers and construction of the genetic high resolution map provide the first steps toward localization of this resistance gene. PMID- 12582492 TI - Microsynteny between the Medicago truncatula SYM2-orthologous genomic region and another region located on the same chromosome arm. AB - A synteny based positional cloning approach was started to clone the pea SYM2 gene by using locally conserved genome structure with the model plant Medicago truncatula. We reported that a pea marker tightly linked to SYM2 was used to screen a M. truncatula BAC library, and two contigs named C1/C2 and C3 were constructed that are both located on the long arm of M. truncatula chromosome 5 and separated by 9 cM. C1/C2 is highly microsyntenic to the pea SYM2 genomic region and corresponds to the M. truncatula SYM2-orthologous region, which is delimitated to 350 kbp. In this manuscript we analyze the distribution in the three contigs of 22 sequences and their homologues, including eight C1/C2 and two pea RFLP markers linked to SYM2. Among the analyzed sequences are several different (receptor) kinase-like gene sequences and two classes of LRR-containing resistance protein-like sequences. From all the studied sequences only four detected homologous sequences in C3, and their distribution is comparable in C1/C2 and C3, suggesting that a 70-kbp and a 120-kbp segments of these two contigs, respectively, arose through a duplication. The implications of these findings for the cloning of SYM2 are discussed. PMID- 12582493 TI - Genetic mapping of grapevine ( Vitis vinifera L.) applied to the detection of QTLs for seedlessness and berry weight. AB - Parental and consensus genetic maps of Vitis vinifera L. (2n = 38) were constructed using a F(1) progeny of 139 individuals from a cross between two partially seedless genotypes. The consensus map contained 301 markers [250 amplification fragment length polymorphisms (AFLPs), 44 simple sequence repeats (SSRs), three isozymes, two random amplified polymorphic DNAs (RAPDs), one sequence-characterized amplified region (SCAR), and one phenotypic marker, berry color] mapped onto 20 linkage groups, and covered 1,002 cM. The maternal map consisted of 157 markers covering 767 cM (22 groups). The paternal map consisted of 144 markers covering 816 cM (23 groups). Differences in recombination rates between these maps and another unpublished map are discussed. The major gene for berry color was mapped on both the paternal and consensus maps. Quantitative trait loci (QTLs) for several quantitative subtraits of seedlessness in 3 successive years were searched for, based on parental maps: berry weight, seed number, seed total fresh and dry weights, seed percent dry matter, and seed mean fresh and dry weights. QTLs with large effects (R(2) up to 51%) were detected for all traits and years at the same location on one linkage group, with some evidence for the existence of a second linked major QTL for some of them. For these major QTLs, differences in relative parental effects were observed between traits. Three QTLs with small effects (R(2) from 6% to 11%) were also found on three other linkage groups, for berry weight and seed number in a single year, and for seed dry matter in 2 different years. PMID- 12582494 TI - Hypomethylation of CNG targets induced with dihydroxypropyladenine is rapidly reversed in the course of mitotic cell division in tobacco. AB - We followed the mitotic transmission of an experimentally induced hypomethylated state of several tobacco repetitive sequences in callus culture and plants. The initial hypomethylation was induced by a hypomethylation drug, dihydroxypropyladenine (DHPA), the competitive inhibitor of cellular S adenosylhomocysteine hydrolase, which is known to preferentially inhibit methylation at CNG and non-symmetrical motifs while having a negligible effect on methylation at CG motifs. The deprivation of this drug resulted in an almost immediate remethylation of cytosines at CNG motifs ( MspI and EcoRII sites) leading us to conclude that, the hypomethylation effect of dihydroxypropyladenine is rather transient and differs from that of 5-azacytidine which often induces heritable changes in methylation patterns. The results suggest that de novo methylation of CNG motifs is a rapid and meiotically independent process on DNA sequences with pre-existing CG methylation. PMID- 12582495 TI - Genetics of resistance to Meloidogyne incognita in crosses of grape rootstocks. AB - Progeny testing was used to investigate the value of selected grape varieties as parents in breeding nematode-resistant rootstocks. Six pistillate-flowered rootstocks (Ramsey, Dog Ridge, Harmony, Freedom, 1613C, and 161-49C) and four staminate-flowered rootstocks (Riparia Gloire, 3309C, 1616C, and St. George) were used. Each male was crossed to each female. Six weeks after inoculation with 1,500 second-stage juveniles of Meloidogyne incognita race 3, roots were stained in an aqueous solution of eosin-Y (0.25 gm/l for 1 h). Seedling resistance was measured by counting the number of stained nematode egg masses visible per root system. Nematode reproduction on each cross was calculated as the average number of egg masses on ten seedlings per replicate. The females Harmony and Freedom produced the greatest level of resistance in their seedlings across all male parents. Seedlings of Dog Ridge, Ramsey, and 1613C had intermediate levels of resistance, while seedlings of 161-49C were the least resistant. The male 1616C contributed the greatest resistance to its progeny, while seedlings from crosses with the males Riparia Gloire, 3309C, and St. George had lower levels of resistance. Segregation ratios of resistant and susceptible seedlings are consistent with a single dominant allele model for root-knot nematode resistance. PMID- 12582496 TI - Chromosome identification and comparative karyotypic analyses of four Pinus species. AB - Chromosomal landmarks in four Pinus species: P. densiflora, P. thunbergii, P. sylvestris, and P. nigra were identified by fluorescence in situ hybridization (FISH) using hapten- or fluorochrome-labeled probes for the plant telomere repeat, centromeric repeat ( PCSR), and rDNA. FISH landmarks were located at the interstitial and proximal regions of chromosomes and allowed us to identify nearly all of the homologous chromosomes in each species. A comparative analysis of the FISH karyotypes among the four species showed that the interstitial FISH signals obtained by hybridization with the telomere and rDNA sequences were stable and could be used to identify homologous chromosomes among species. The identification of homologous chromosomes among species facilitated a detailed comparative karyotype analysis. The results suggest that the degree of chromosomal differentiation among the four Pinus species is very low and that the proximal regions vary in their DNA sequences. The similarities and differences among FISH karyotypes are discussed in relation to phylogeny. PMID- 12582497 TI - Arabidopsis thaliana-derived resistance against Leptosphaeria maculans in a Brassica napus genomic background. AB - Stem canker (blackleg) caused by Leptosphaeria maculans is a widespread disease of Brassica napus. In contrast, most Arabidopsis thaliana accessions are highly resistant. Hence, novel material derived from symmetric and asymmetric somatic hybrids between B. napus and A. thaliana was utilised in a screen for L. maculans resistance. Initially, both cotyledon and adult-leaf resistance traits were transferred from A. thaliana to B. napus. In later generations the two traits segregated and cotyledon resistance was lost. The adult-leaf resistance was investigated with respect to genome localisation and protein expression. Analyses of remaining A. thaliana DNA in resistant plants showed co-segregation between adult-leaf resistance and chromosome-3 molecular markers. Resistant offspring from asymmetric hybrid plants that contained fragments of chromosome 3 were studied in more detail. Two regions at positions 9.8-10.4 Mbp and 18-19.5 Mbp, where several defence-related genes are located, were identified. A proteomic approach was taken to further investigate genes involved in the defence interaction. Forty eight hours after inoculation with L. maculans, only a few proteins, such as glycolate oxidase, were identified as differentially expressed in the resistant line compared to B. napus, despite the presence of additional A. thaliana chromosomes. The plant materials described in the present study constitute a new genetic source of L. maculans resistance and are currently being incorporated into B. napus breeding programmes. PMID- 12582498 TI - A constitutive gene expression system derived from the tCUP cryptic promoter elements. AB - A limited number of constitutive promoters have been used to direct transgene expression in plants and they are often derived from non-plant sources. Here, we describe novel gene-regulatory elements which are associated with a cryptic constitutive promoter from tobacco, tCUP, and modifications that were made to create a strong gene-expression system that is effective across all living cell types from a wide range of plant species, including several important crops ( Arabidopsis, canola, flax, alfalfa, tobacco). The tCUP 5' untranslated region was mutated to eliminate translational interference by upstream ATGs, and the influence of the Kozak consensus sequence on the levels of a beta-glucuronidase (GUS) reporter gene activity was demonstrated. These modifications resulted in expression that was greatly enhanced in all organs. A TATA consensus sequence was added to the core promoter to complement an existing Initiator (Inr) sequence. Although this addition was known to elevate core promoter activity by 3-fold the additive effect on the overall gene-expression system was marginal in all of the transgenic plants tested. Two transcriptional enhancers were identified and the region containing them were oligomerized, yielding a significant increase in marker gene-expression in some but not all plant species. In general, the enhanced tCUP gene-expression system generated levels of GUS activity which exceeded that of the 35S promoter in most plant species and the elevation in activity occurred uniformly among the various plant organs. The potential benefit of cryptic elements for the construction of gene-expression systems for crop species is discussed PMID- 12582499 TI - Enhancement of metabolizing herbicides in young tubers of transgenic potato plants with the rat CYP1A1 gene. AB - A rat P450 monooxygenase gene ( CYP1A1) was introduced into potato plants to enhance the metabolism of the environmental contaminants in subterranean organs. The CYP1A1 gene was kept under the control of the potato patatin promoter to enhance tuber-specific expression. A total of 106 transgenic plants (PAT1A1 plants) were obtained following selection by a resistance test to kanamycin and PCR analysis. PAT1A1 plants treated with 10% exogenous sucrose showed a higher activity of monooxgenase in the leaves than the non-transgenic plants. This indicated that the activity enhanced by 10% sucrose was due to the patatin promoter containing the sucrose-inducted elements. One representative transgenic plant, Ag2197, was selected on the basis of monooxgenase activity in the leaves and Western blot analysis. Ag2197 was found to accumulate a large amount of CYP1A1 mRNA and protein in the developing tuber but not in the mature tuber. The residual herbicides, atrazine and chlortoluron, were analyzed in the micro-tubers of Ag2197 and non-transgenic plants. The amount of residual herbicides in Ag2197 was much lower than that in the non-transgenic plant, indicating that the transgenic plant metabolized the herbicides to a detoxified form. The transgenic plants produced in this study might be useful for the phytoremediation of chemical pollution in the soil. PMID- 12582500 TI - Genetic mapping of a dominant gene conferring resistance to cassava mosaic disease. AB - Cassava mosaic disease (CMD) is the most-important disease of cassava ( Manihot esculenta) in Africa, and is a potential threat to Latin American (LA) cassava production. Although this viral disease is still unknown in LA, its vector - the whitefly - has recently been found. The disease is best controlled through host plant resistance, which was first found in third backcross derivatives of an interspecific cross between cassava and Manihot glaziovii, and is thought to be polygenic. Recently, high levels of resistance were also found in several Nigerian cassava landraces. Classical genetic analysis and molecular genetic mapping of the landraces showed that a major dominant gene confers this resistance. Bulk segregant analysis (BSA) was used to quickly identify a simple sequence repeat (SSR) marker linked to the CMD-resistance gene. The marker, SSRY28, is located on linkage group R of the male-parent-derived molecular genetic map. The gene, designated as CMD2, is flanked by the SSR and RFLP marker GY1 at 9 and 8 cM, respectively. To our knowledge, this is the first report of qualitative virus resistance in cassava, and of molecular markers that tag CMD resistance in cassava. We discuss the use of markers linked to CMD2 for marker assisted breeding of CMD resistance in Latin America and for increasing the cost effectiveness of resistance breeding in Africa. PMID- 12582501 TI - Genetic analysis of growth curves for a woody perennial species, Pinus taeda L. AB - Inheritance of growth curves is critical for understanding evolutionary change and formulating efficient breeding plans, yet has received limited attention. Growth curves, like other characters that change in concert with development, often have higher heritability than age-specific traits. This study compared genetic parameters of height-growth curves with those of age-specific heights for a conifer, Pinus taeda L. Growth curves were fitted with: (1) a linear regression model, and (2) a non-linear model based on Richards' function using two sources of height data: two six-parent diallel tests assessed at age 2 to 10 years and two tests from a nested mating design with 222 parents assessed at 1 to 25 years. Additive genetic control of growth-curve parameters was moderate (h(2) = 0.06 to 0.26) and slightly lower than that for age-specific heights. Additive variance exceeded dominance variance for rate and shape parameters, but not for the asymptote. Genetic correlations among growth-curve parameters were high. Early selection on height was as efficient as selection on growth-curve parameters. PMID- 12582502 TI - Characterization of the flanking regions of Zea mays microsatellites reveals a large number of useful sequence polymorphisms. AB - Sequence characterization of the flanking regions of 52 sequence-tagged microsatellite loci and two gene fragments from 11 Zea mays inbred lines identified a total of 324 sequence polymorphisms. The sequence polymorphisms consisted of both single-nucleotide polymorphisms and insertions/deletions in a ratio of approximately two to one. The level of sequence variation within the flanking regions of microsatellites linked to expressed sequence tags was lower than microsatellites that were unlinked to expressed sequence tags. However, both types of microsatellites generated a similar number of sequence-based alleles across the 11 genotypes surveyed. In two out of 20 microsatellites examined in detail, evidence was found for size-based allele homoplasy. Conversion of the observed sequence polymorphisms into allele-specific oligonucleotides followed by covalent binding to glass slides allowed the sequence polymorphisms to be used in a simple hybridization-based genotyping procedure. This procedure enabled us to discriminate between different inbred lines and allowed variations within a single inbred to be identified. The sequence information presented in this report could be used as a starting point for other programmes in the further development of a non-gel based, multi-locus, multi-allele screen for large-scale maize genotyping. PMID- 12582503 TI - Markers polymorphic among malting barley ( Hordeum vulgare L.) cultivars of a narrow gene pool associated with key QTLs. AB - Barley used for malting is a fine-tuned organism, and it requires breeding within narrow gene pools for realistic cultivar enhancement. Significant phenotypic advance within such narrow gene pools has been achieved and the necessary genetic variability for breeding progress has been documented, but it was not well understood. This study was conducted to further characterize detectable genetic variability present within a select set of four closely related malting barley cultivars using three types of molecular markers: RFLP, PCR-RAPD and AFLP. The markers that identified polymorphism among the select malting cultivars tended to link with each other and to map in chromosomal regions associated with quantitative trait loci (QTLs) for agronomic and malting quality traits that differed among the four cultivars. Although RFLPs identified the least amount of polymorphism, the differences detected by the RFLPs best fit the chronology of the cultivars. These results indicate that a large amount of the genetic variability necessary for cultivar improvement may have originally been present in the breeding gene pool, but does not rule out de novo variation. Study of the populations from crosses within this narrow germplasm is needed to further elucidate the basis of the phenotypic variability found among these select barley cultivars. PMID- 12582504 TI - A genetic linkage map for watermelon derived from a testcross population: ( Citrullus lanatus var. citroides x C. lanatus var. lanatus) x Citrullus colocynthis. AB - A genetic linkage map was constructed for watermelon using a testcross population [Plant Accession Griffin 14113 ( Citrullus lanatus var. citroides) x New Hampshire Midget (NHM; C. lanatus var. lanatus)] x U.S. Plant Introduction (PI) 386015 ( Citrullus colocynthis). The map contains 141 randomly amplified polymorphic DNA (RAPD) markers produced by 78 primers, 27 inter-simple sequence repeat (ISSR) markers produced by 17 primers, and a sequence-characterized amplified region (SCAR) marker that was previously reported as linked (1.6 cM) to race-1 Fusarium wilt [incited by Fusarium oxysporum Schlechtend.:Fr. f. sp. niveum (E.F.Sm.) W.C. Synder & H.N. Hans] resistance in watermelon. The map consists of 25 linkage groups. Among them are a large linkage group that contains 22 markers covering a mapping distance of 225.6 cM and six large groups each with 10-20 markers covering a mapping distance of 68.8 to 110.8 cM. There are five additional linkage groups consisting of 3-7 markers per group, each covering a mapping distance of 36.5 to 57.2 cM. The 13 remaining linkage groups are small, each consisting of 2-11 markers covering a mapping distance of 3.5-29.9 cM. The entire map covers a total distance of 1,166.2 cM with an average distance of 8.1 cM between two markers. This map is useful for the further development of markers linked to disease resistance and watermelon fruit qualities. PMID- 12582505 TI - The Potato virus S resistance gene Ns maps to potato chromosome VIII. AB - The dominant allele Ns confers in potato resistance to Potato virus S (PVS). To identify the chromosomal location of Ns, we mapped the Ns-linked marker SCG17(448) and the ISSR marker UBC811(600) to linkage group VIII of the RFLP map of a population that did not segregate for Ns. The map position of the Ns locus on chromosome VIII was confirmed with the detection of linkage between Ns and three RFLP markers, GP126, GP189 and CP16, known to be located in a corresponding region on potato chromosome VIII. PCR-based assays were developed for these RFLP markers. The PCR primers specific for GP126 generated polymorphic products (STS marker). In the case of markers GP189 and CP16, informative polymorphism was revealed in the Ns population after digestion with the restriction enzymes HaeIII and HindIII, respectively. The genetic distance between Ns and the closest CP16 locus was 4.2 cM. PMID- 12582506 TI - Identification of genetic factors controlling the efficiency of Agrobacterium rhizogenes-mediated transformation in Brassica oleracea by QTL analysis. AB - We have identified quantitative trait loci (QTL) for transgenic and adventitious root production using an Agrobacterium rhizogenes-mediated co-transformation system in conjunction with a Brassica oleracea double haploid (DH) mapping population. Three QTL for green fluorescent protein (GFP)-fluorescent root production and four QTL for adventitious root production were identified as accounting for 26% and 32% of the genetic variation in the population, respectively. Two of the QTL regions identified were common to both transgenic and adventitious root production. Two different methods of QTL analysis were employed (marker regression and interval mapping) and with the exception of one region on linkage group O7 for transgenic root production, both techniques detected the same regions of the genome. The regions we identified to be associated with the control of transgenic root production following A. rhizogenes mediated transformation are the first to be detected using a QTL mapping approach. In addition, this is the first study to identify genetic regions that co-regulate both transgenic and adventitious root production within the constraints of an A. rhizogenes-mediated transformation process. We have identified plant genotypes that do not produce any transgenic roots that may be deficient for T-DNA integration via illegitimate recombination and that may also be potentially important for the development of homologous recombination protocols. Conversely, we have also identified plant genotypes with high rates of transgenic root production that will be critical in the development of high throughput transformation systems. PMID- 12582507 TI - An SSR-based genetic linkage map for perennial ryegrass ( Lolium perenne L.). AB - A simple sequence repeat (SSR)-based linkage map has been constructed for perennial ryegrass ( Lolium perenne L.) using a one-way pseudo-testcross reference population. A total of 309 unique perennial ryegrass SSR (LPSSR) primer pairs showing efficient amplification were evaluated for genetic polymorphism, with 31% detecting segregating alleles. Ninety-three loci have been assigned to positions on seven linkage groups. The majority of the mapped loci are derived from cloned sequences containing (CA)(n)-type dinucleotide SSR arrays. A small number (7%) of primer pairs amplified fragments that mapped to more than one locus. The SSR locus data has been integrated with selected data for RFLP, AFLP and other loci mapped in the same population to produce a composite map containing 258 loci. The SSR loci cover 54% of the genetic map and show significant clustering around putative centromeric regions. BLASTN and BLASTX analysis of the sequences flanking mapped SSRs indicated that a majority (84%) are derived from non-genic sequences, with a small proportion corresponding to either known repetitive DNA sequence families or predicted genes. The mapped LPSSR loci provide the basis for linkage group assignment across multiple mapping populations. PMID- 12582508 TI - Mapping of a thermo-sensitive earliness per se gene on Triticum monococcum chromosome 1A(m). AB - An earliness per se gene, designated Eps-A(m) 1, was mapped in diploid wheat in F(2) and single-seed descent mapping populations from the cross between cultivated (DV92) and wild (G3116) Triticum monococcum accessions. A QTL with a peak on RFLP loci Xcdo393 and Xwg241, the most distal markers on the long arm of chromosome 1A(m), explained 47% of the variation in heading date (LOD score 8.3). Progeny tests for the two F(2:3) families with critical recombination events between Xcdo393 and Xwg241 showed that the gene was distal to Xcdo393 and linked to Xwg241. Progeny tests and replicated experiments with line #3 suggested that Eps-A(m) 1 was distal to Xwg241. This gene showed a large effect on heading date in the controlled environment experiments, and a smaller, but significant, effect under natural conditions. Eps-A(m) 1 showed significant epistatic interactions with photoperiod and vernalization treatments, suggesting that the different classes of genes affecting heading date interact as part of a complex network that controls the timing of flowering induction. Besides its interactions with other genes affecting heading date, Eps-A(m) 1 showed a significant interaction with temperature. The effect of temperature was larger in plants carrying the DV92 allele for late flowering than in those carrying the G3116 allele for early flowering. Average differences in heading date between the experiments performed at 16 degrees C and 23 degrees C were approximately 11 days ( P < 0.001) for the lines carrying the Eps-A(m) 1 allele for early flowering but approximately 50 days ( P < 0.0001) for the lines carrying the allele for late flowering. The large differences in heading time (average 80 days) observed between plants carrying the G3116 and DV92 alleles when grown at 16 degrees C, suggest that it would be possible to produce very detailed maps for this gene to facilitate its future positional cloning. PMID- 12582509 TI - Endogenous enzyme activities and polyamine levels in diverse rice cultivars depend on the genetic background and are not affected by the presence of the hygromycin phosphotransferase selectable marker. AB - We used the polyamine biosynthetic pathway and rice as a relevant model to understand the genetic basis of variation in endogenous levels of metabolites and key enzymes involved in the pathway. Wild-type tissues and also tissues containing a commonly used selectable marker gene were employed. We detected a wide variation in levels of arginine decarboxylase activity and in the three polyamines, putrescine, spermidine and spermine, in different tissues and varieties, but this was not dependent on the presence of the selectable marker. A more-extensive profile of enzyme activities (ADC, ODC, SAMDC, DAO and PAO) and polyamine levels in different tissues was generated in two different varieties. Our results indicate that genetic background is important in terms of the basal levels of metabolites and enzyme activity, particularly in situations in which we aim to engineer metabolic pathways that are also encoded by homologous endogenous genes. We did not find any evidence that the presence of a selectable marker in any way influences enzyme activity or metabolite levels. PMID- 12582510 TI - Identification of an STMS marker for the double-podding gene in chickpea. AB - Chickpea, a self-pollinating diploid annual with 2n = 2 x = 16 chromosomes, is an important food legume crop throughout the world and especially in developing countries. A gene that confers double-podding, with the symbol " s", is considered important for breeding higher yielding cultivars. Positive effects of " s" on yield stability were found by comparing single- and double-podded near isogenic lines (NILs) derived from a cross of CA-2156 (single-podded) with JG-62 (double-podded). Considering the significant effects on seed yield, the NILs were used to identify molecular markers closely linked to " s". Sequence tagged microsatellite site (STMS) markers developed for chickpea, random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers were used to analyze the NILs. Out of 400 RAPD, 100 STMS and 100 ISSR markers, one STMS marker (TA-80) was polymorphic and was used to evaluate a recombinant inbred line population developed from the cross of Surutato-77 (single-podded) x JG-62 (double-podded) for co-segregation of the locus with " s". Our results indicated that the marker and " s" were 4.84 cM apart. This marker may be used by breeders for marker-assisted selection (MAS) to monitor the incorporation of the double podding gene into improved germplasm. PMID- 12582511 TI - Origin of cytoplasm substituted rice cultivars found in Japan. AB - Genetic variation of Japanese rice cultivars were examined. Five of 450 lowland cultivars and another five of 200 upland cultivars were determined as the indica type by using isozyme genotypes and the remainder were of the japonica type. The major characteristics of these indica cultivars, revealed a slender shape of grains, a short apiculus hair length, a positive allele for Ph reaction, and allele-3 for the Pgd1 locus. Three of these indica cultivars showed a non deletion ORF100, which is essential to the japonica-type plastid. The plastid subtype identity (PS-ID) sequences of these plastids is 6C7A, which is also a japonica-specific repeat unit. Thus, these cultivars were concluded to be naturally generated cytoplasm substituted lines. These plastids were introduced into a indica genetic background from japonica cultivars grown elsewhere. The rest of the indica cultivars revealed a deletion-type ORF100 and plastid subtype 8C8A, both of which are indica-specific. These cultivars carried indica-type allelic constitutions for diagnostic isozyme loci. However, other characters were identical to the cytoplasm-substituted cultivars in Japan. In East and Southeast Asia, cultivars carrying a indica-type nuclear genotype with a japonica-type plastid are restricted to Aus cultivars in the Bengal region. Genetic and historical records suggest that Japanese indica cultivars and the Aus cultivars are closely related. The Aus cultivars acquire necessary genetic constitutions from both indica and japonica cultivars through naturally occurring out-crossing to adapt to a particular cultivation condition in the region. The wide adaptability enabled them to be introduced into a northern region like Japan. PMID- 12582512 TI - Complete sequence of Tvv1, a family of Ty 1 copia-like retrotransposons of Vitis vinifera L., reconstituted by chromosome walking. AB - A chromosome-walking strategy was used to sequence and characterize retrotransposons in the grapevine genome. The reconstitution of a family of retroelements, named Tvv1, was achieved by six successive steps. These elements share a single, highly conserved open reading frame 4,153 nucleotides-long, putatively encoding the gag, pro, int, rt and rh proteins. Comparison of the Tvv1 open reading frame coding potential with those of drosophila copia and tobacco Tnt1, revealed that Tvv1 is closely related to Ty 1 copia-like retrotransposons. A highly variable untranslated leader region, upstream of the open reading frame, allowed us to differentiate Tvv1 variants, which represent a family of at least 28 copies, in varying sizes. This internal region is flanked by two long terminal repeats in direct orientation, sized between 149 and 157 bp. Among elements theoretically sized from 4,970 to 5,550 bp, we describe the full-length sequence of a reference element Tvv1-1, 5,343 nucleotides-long. The full-length sequence of Tvv1-1 compared to pea PDR1 shows a 53.3% identity. In addition, both elements contain long terminal repeats of nearly the same size in which the U5 region could be entirely absent. Therefore, we assume that Tvv1 and PDR1 could constitute a particular class of short LTRs retroelements. PMID- 12582513 TI - Chromosomal regions associated with segregation distortion in maize. AB - Segregation distortion skews the genotypic frequencies from their Mendelian expectations. Our objectives in this study were to assess the frequency of occurrence of segregation distortion in maize, identify chromosomal regions consistently associated with segregation distortion, and examine the effects of gametophytic factors on linkage mapping. We constructed a simple sequence repeat (SSR) linkage map for a LH200/LH216 F(2)Syn3 (i.e., random-mated three times) population, and compared the segregation distortion in this map with the segregation distortion in three published linkage maps. Among 1,820 codominant markers across the four mapping populations, 301 (17%) showed segregation distortion ( P < 0.05). The frequency of markers showing segregation distortion ranged from 19% in the Tx303/CO159 mapping population to 36% in the B73/Mo17 mapping population. A positive relationship was found between the number of meioses and the frequency of segregation distortion detected in a population. On a given chromosome, nearly all of the markers showing segregation distortion favored the allele from the same parent. A total of 18 chromosomal regions on the ten maize chromosomes were associated with segregation distortion. The consistent location of these chromosomal regions in four populations suggested the presence of segregation distortion regions (SDRs). Three known gametophytic factors are possible genetic causes of these SDRs. As shown in previous research, segregation distortion does not affect the estimate of map distance when only one gametophytic factor is present in an SDR. PMID- 12582514 TI - Construction of a combined sorghum linkage map from two recombinant inbred populations using AFLP, SSR, RFLP, and RAPD markers, and comparison with other sorghum maps. AB - Sorghum [ Sorghum bicolor (L.) Moench] is an important crop in the semi-arid tropics that also receives growing attention in genetic research. A comprehensive reference map of the sorghum genome would be an essential research tool. Here, a combined sorghum linkage map from two recombinant inbred populations was constructed using AFLP, SSR, RFLP and RAPD markers. The map was aligned with other published sorghum maps which are briefly reviewed. The two recombinant inbred populations (RIPs) analyzed in this study consisted of 225 (RIP 1) and 226 (RIP 2) F(3:5) lines, developed from the crosses IS 9830 x E 36-1 (RIP 1) and N 13 x E 36-1 (RIP 2), respectively. The genetic map of RIP 1 had a total length of 1,265 cM (Haldane), with 187 markers (125 AFLPs, 45 SSRs, 14 RFLPs, 3 RAPDs) distributed over ten linkage groups. The map of RIP 2 spanned 1,410 cM and contained 228 markers (158 AFLPs, 54 SSRs, 16 RFLPs) in 12 linkage groups. The combined map of the two RIPs contained 339 markers (249 AFLPs, 63 SSRs, 24 RFLPs, 3 RAPDs) on 11 linkage groups and had a length of 1,424 cM. It was in good agreement with other sorghum linkage maps, from which it deviated by a few apparent inversions, deletions, and additional distal regions. PMID- 12582515 TI - Genetic diversity and relationships in olive ( Olea europaea L.) germplasm collections as determined by randomly amplified polymorphic DNA. AB - Genetic diversity studies using the RAPD technique were carried out in a set of 103 olive cultivars from the World Germplasm Bank of the Centro de Investigacion y Formacion Agraria (CIFA) "Alameda del Obispo" in Cordoba (Spain). A total of 126 polymorphisms (6.0 polymorphic markers per primer) out of 135 reproducible products (6.4 fragments per primer) were obtained from the 21 primers used. The number of bands per primer ranged from 4 to 11, whereas the number of polymorphic bands ranged from 3 to 10, corresponding to 83% of the amplification products. The dendrogram based on unweighted pair-group cluster analysis using Jaccard's index includes three major groups according to their origin: (1) cultivars from the Eastern and Central Mediterranean areas, (2) some Italian and Spanish cultivars, and (3) cultivars from the Western Mediterranean zone. The pattern of genetic variation among olive cultivars from three different Mediterranean zones (West, Centre and East) was analysed by means of the analysis of molecular variance (AMOVA). Although most of the genetic diversity was attributable to differences of cultivars within Mediterranean zones (96.86%) significant phi values among zones (phi(st) = 0.031; p < 0.001) suggested the existence of phenotypic differentiation. Furthermore, the AMOVA analysis was used to partition the phenotypic variation of Spain, Italy (Western region), Greece and Turkey (Eastern region) into four categories: among regions, among countries (within regions), within countries, and among and within countries of each region. Most of the genetic diversity was attributable to differences among genotypes within a country. These results are consistent with the predominantly allogamous nature of Olea europaea L. species. This paper indicates the importance of the study of the amount and distribution of genetic diversity for a better exploration of olive genetic resources and the design of plant breeding programmes. PMID- 12582516 TI - Evolution of 5S rDNA units and their chromosomal localization in Allium cepa and Allium schoenoprasum revealed by microdissection and FISH. AB - Allium cepa and Allium schoenoprasum each possess 5S rDNA units of two different sizes. The evolution of the two repeat units and their chromosomal localization were investigated. A. cepa has 5S rDNA loci in the proximal and distal regions of the short arm of chromosome 7. When the proximal and distal segments of the short arm of chromosome 7 were microdissected separately, and used as templates for PCR, the short and long 5S rDNA fragments were amplified predominantly from the proximal and distal segments, respectively. The nucleotide sequence of the long 5S rDNA unit resulted from partial duplication of a non-transcribed spacer (NTS) and the insertion of a unique sequence. FISH using a probe consisting of the unique sequence demonstrated that the long unit was distally localized. In A. cepa, the long 5S rDNA unit is only present distally and the short unit is predominantly located proximally on the short arm of chromosome 7. In A. schoenoprasum, the NTSs of the two different-sized 5S rDNAs had quite different sequences. The two 5S rDNA loci were localized very close together in the interstitial region of chromosome 6. FISH, using long and short 5S rDNA unit probes with a competitor of a 120-bp sequence of the 5S rRNA gene, indicated that the long 5S rDNA unit was localized proximally and the short unit distally. Although the NTSs of the 5S rDNA of A. cepa and A. schoenoprasum had quite different nucleotide sequences, the long 5S rDNA units of A. cepa and A. schoenoprasum share a common 75-bp sequence. This sequence might act in the formation of the long 5S rDNA unit in Allium species. PMID- 12582517 TI - Transfer of a male-sterility-inducing cytoplasm from onion to leek ( Allium ampeloprasum). AB - Two interspecific triploid (AAC) hybrids (84/1-94 and 99/1-94) from crosses between onion [ Allium cepa (2 n=2 x=16, CC)] and leek [ A. ampeloprasum (2 n=4 x=32, AAAA)] were backcrossed to leek in order to transfer a male-sterility inducing cytoplasm from onion that would enable the production of hybrid leek. GISH evaluations of meiosis in the interspecific hybrids revealed irregularities due to univalent onion chromosomes producing micronuclei from onion chromatin, whereas the pairing of the two sets of leek chromosomes was nearly normal. Attempts to use colchicine to double the chromosome number of the hybrids failed. Backcrosses of 84/1-94 to leek as the pollen parent were not successful. The first backcross of 99/1-94 to tetraploid leek produced 11 BC(1) plants with chromosome numbers between 38 and 41. Identification of parental chromosomes by GISH showed that all eight onion chromosomes and 30-33 leek chromosomes were transmitted to the backcross progenies due to unreduced egg cells. Onion chromosomes were eliminated during the second backcross. Southern hybridization confirmed the transfer of the T-cytoplasm like source of CMS from onion to the BC(2) progenies. After the third backcross to leek, 158 plants were obtained with varying numbers of onion chromosomes and some intergenomic recombinant chromosomes. Alloplasmic leek plants without onion chromatin were selected for further characterization of male sterility and quality traits. PMID- 12582518 TI - Genetic linkage maps of two apricot cultivars ( Prunus armeniaca L.), and mapping of PPV (sharka) resistance. AB - Genetic linkage maps for two apricot cultivars have been constructed using AFLP, RAPD, RFLP and SSR markers in 81 F1 individuals from the cross 'Goldrich' x 'Valenciano'. This family segregated for resistance to 'plum pox virus' (PPV), the most-important virus affecting Prunus species. Of the 160 RAPD arbitrary primers screened a total of 44 were selected. Sixty one polymorphic RAPD markers were scored on the mapping population: 30 heterozygous in 'Goldrich', 19 heterozygous in 'Valenciano', segregating 1:1, and 12 markers heterozygous in both parents, segregating 3:1. A total of 33 and 19 RAPD markers were mapped on the 'Goldrich' and 'Valenciano' maps respectively. Forteen primer combinations were used for AFLPs and all of them detected polymorphism. Ninety five markers segregating 1:1 were identified, of which 62 were heterozygous in the female parent 'Goldrich' and 33 in the male parent 'Valenciano'. Forty five markers were present in both parents and segregated 3:1. A total of 82 and 48 AFLP markers were mapped on the 'Goldrich' and 'Valenciano' maps. Twelve RFLPs probes were screened in the population, resulting in five loci segregating in the family, one locus heterozygous for 'Valenciano' and four heterozygous for both, segregating 1:2:1. Of the 45 SSRs screened 17 segregated in the mapping family, resulting in seven loci heterozygous for the maternal parent and ten heterozygous for both, segregating 1:2:1 or 1:1:1:1. A total of 16 and 13 co-dominant markers were mapped in the female and male parent maps respectively. A total of 132 markers were placed into eight linkage groups on the 'Goldrich' map, defining 511 cM of the total map-length. The average distance between adjacent markers was 3.9 cM. A total of 80 markers were placed into seven linkage groups on the 'Valenciano' map, defining 467.2 cM of the total map-distance, with an average interval of 5.8 cM between adjacent markers. Thirty six marker loci heterozygous in both parents revealed straightforward homologies between five linkage groups in both maps. The sharka resistance trait mapped on linkage group 2. The region containing sharka resistance is flanked by two co-dominant markers that will be used for targeted SSR development employing a recently constructed complete apricot BAC library. SSRs tightly linked to sharka resistance will facilitate MAS in breeding for resistance in apricot. PMID- 12582519 TI - High-resolution genetic map of Nb, a gene that confers hypersensitive resistance to potato virus X in Solanum tuberosum. AB - Nb is a single dominant gene in potato that confers hypersensitive resistance to potato virus X (PVX) isolates from strain groups 1 and 2. Genetic and molecular analyses showed that Nb is located on the upper arm of chromosome V and forms part of a cluster of resistance genes encoding specificities to many different pathogens. We describe the genetical localisation of molecular markers tightly linked to the Nb locus and the development PCR-based markers suitable for isolation of the Nb resistance gene by positional cloning. A bulked segregant approach was applied to identify polymorphic AFLP markers tightly linked to the Nb locus. These markers were mapped in a population of segregating S1 progeny (1,300 plants) from a self-pollinated potato cultivar, Pentland Ivory. From this analysis, Nb was placed in an interval of 0.76 cM, flanked by the AFLP markers GM339 and GM637. Recombinant PVX strains carrying different combinations of avirulence genes were used in biological assays to show that Nb was also present in potato cv. Cara but was masked by the extreme PVX resistance conferred by the Rx gene. PCR-based screening of a Cara genomic BAC library with markers closest to the Nb locus identified a new marker tightly linked to Nb. PMID- 12582520 TI - Competence of oat ( Avena sativa L.) shoot apical meristems for integrative transformation, inherited expression, and osmotic tolerance of transgenic lines containing hva1. AB - Three oat ( Avena sativa L.) cultivars have been successfully transformed using an efficient and reproducible in vitro culture system for differentiation of multiple shoots from shoot apical meristems. The transformation was performed using microprojectile bombardment with two plasmids (pBY520 and pAct1-D) containing linked ( hva1-bar) and non-linked ( gus) genes. The hva1 and bar genes cointegrated with a frequency of 100% as expected, and 61.6% of the transgenic plants carried all three genes. Molecular and biochemical analyses in R0, R1 and R2 progenies confirmed stable integration and expression of all transgenes. Localization of the GUS protein in R0 and R1 plants revealed that high-expression of gus occurred in vascular tissues and in the pollen grains of mature flowers. The constitutive expression of HVA1 protein was observed at all developmental stages of transgenic plants, and was particularly stronger during the early seedling stages. R2 progeny of five independent transgenic lines was tested in vitro for tolerance to osmotic (salt and mannitol) stresses. As compared to non transgenic control plants, transgenic plants maintained a higher growth and showed significantly ( P < 0.05) increased tolerance to stress conditions. Less than 10% of transgenic plants showed symptoms of wilting or death of leaves and, when these symptoms present were delayed in transgenic plants as compared to 80% of non-transgenic plants, either wilted or died. These symptoms confirmed the increased in vitro tolerance in hva1-expressing transgenic plants to non transgenic plants, providing strong evidence that the HVA1 protein may play an important role in the protection of oats against salinity and possible water deficiency stress conditions. PMID- 12582521 TI - AFLP analysis of genetic diversity within and among Coffea arabica cultivars. AB - Genetic diversity of Coffea arabica cultivars was estimated using amplified fragment length polymorphism (AFLP) markers. Sixty one Coffea accessions composed of six arabica cultivars, including Typica, Bourbon, Catimor, Catuai, Caturra and Mokka Hybrid, plus two diploid Coffea species, were analyzed with six EcoRI- MseI primer combinations. A total of 274 informative AFLP markers were generated and scored as binary data. These data were analyzed using cluster methods in the software package NTSYSpc. The differences among cultivars at the DNA level were small, with an average genetic similarity of 0.933. Most accessions within a cultivar formed a cluster, although deviant samples occurred in five of the six cultivars examined due to residual heterozygosity from ancestral materials. Among the six cultivars fingerprinted, the highest level of genetic diversity was found within the cultivar Catimor, with an average genetic similarity of 0.880. The lowest level was found within Caturra accessions, with an average genetic similarity of 0.993. Diversity between C. arabica and two other Coffea species, Coffea canephora and Coffea liberica, was also estimated with average genetic similarities of 0.540 and 0.413, respectively, suggesting that C. canephora is more closely related to C. arabica than is C. liberica. The genetic variation among arabica cultivars was similar to the variation within cultivars, and no cultivar-specific DNA marker was detected. Although arabica cultivars appear to have a narrow genetic base, our results show that sufficient polymorphism can be found among some arabica cultivars with a genetic similarity as low as 0.767 for genetic/QTL mapping and breeding. The assessment of genetic diversity among arabica cultivars provided the necessary information to estimate the potential for using marker-assisted breeding for coffee improvement. PMID- 12582522 TI - Cloning of cDNA for UDP-glucose pyrophosphorylase and the expression of mRNA in rice endosperm. AB - Rice endosperm UDP-glucose pyrophosphorylase (UGPase) cDNA clones were isolated by screening a lambda ZAP II library prepared from poly (A(+)) RNA of japonica rice (cv Sasanishiki) endosperm with a probe of potato UGPase cDNA. One cDNA clone, possessing about 1,700 nucleotides, contained the complete open reading frame of rice UGPase. At the nucleotide-sequence level, the UGPase cDNA of rice endosperm had high homology with the UGPase cDNA of barley endosperm (84%) and potato tuber (71%). The calculated molecular weight (50 kDa) agrees with the value determined by SDS-PAGE (51 kDa). At the amino-acid sequence level, rice UGPase has high homology with the UGPase of barley (92%) and potato (85%). The enzyme contained conserved sequence elements which are thought to be involved in substrate binding and catalytic activity. A Southern-blot analysis indicated that the gene existed as a single copy. Expression of the enzyme in rice endosperm examined by Northern-blot analysis was high at 10-15 days after heading. PMID- 12582523 TI - Inheritance of S(f)-RNase in Japanese apricot ( Prunus mume) and its relation to self-compatibility. AB - Self-compatible cultivars of Japanese apricot ( Prunus mume Shieb. et Zucc.), a tree species that normally shows S-RNase-based self-incompatiblity, have a horticultural advantage over self-incompatible cultivars. Inheritance of self compatibility and a common S(f)-RNase allele that is observed in self-compatible cultivars was investigated using progenies from controlled crosses. Total DNAs were isolated from the parents and progenies of seven crosses that included at least one self-compatible cultivar as a parent. These DNAs were PCR-amplified with the Pru-C2 and PCE-R primer pair to determine S-haplotypes of the parents and progenies. A novel S-haplotype, S(8), was found. In all crosses examined, the S(f)-RNase gene was inherited from either the seed or pollen parent as a pistil S allele in a non-functional S-haplotype. Self-compatibility of about 20 trees each from reciprocal crosses of 'Benisashi ( S(7) S(f))' and 'Shinpeidayu ( S(3) S(f))', and 26 selections from 16 different crosses was tested by pollination and pollen-tube growth studies. Cosegregation of the S(f)-RNase allele and self compatibility was confirmed with all but selection 1K0-26 ( S(3) S(7)). Selection 1K0-26 ( S(3) S(7)) that originated from 'Benisashi ( S(7) S(f))' x 'Koshinoume ( S(3) S(f))' appeared to be self-compatible even without the S(f)-RNase allele. The possible role of pollen- S, a presumably existing pollen component of gametophytic self-incompatibility, is discussed. PMID- 12582524 TI - Genetic diversity of Eritrean sorghum landraces assessed with simple sequence repeat (SSR) markers. AB - A precise high-throughput approach was used to characterize diversity in 28 Eritrean sorghum landraces, and to compare this diversity to representative samples of the world sorghum collection. Pools of simple sequence repeat (SSR) markers were sized and scored on automated DNA-sizing gels. An exceptionally high level of diversity was observed among the 28 Eritrean landraces, compared to other sorghum germplasms, in both the number and size range of SSR alleles. Individual landraces were found to carry a high level of within-population diversity and heterozygosity, and between-population diversity was equally high. Eritrean sorghums could be clustered into 7-10 major subgroups, with most (but not all) classifications agreeing with descriptions by farmers. Clustering did not concur particularly well with the major classification system applied in Eritrea, highland versus lowland. Eight of the Eritrean landraces grouped with other sorghums in the world collection, particularly those from Ethiopia/Sudan and India or of the durra and caudatum races, but most Eritrean sorghums clustered in a separate subgroup. These results indicate that a great deal of germplasm diversity and genetic novelty are available in Eritrean sorghums, and that SSR markers can contribute to the wise use of this diversity for sorghum study and improvement. PMID- 12582525 TI - QTLs affecting kernel size and shape in a two-rowed by six-rowed barley cross. AB - The suitability of barley ( Hordeum vulgare L.) grain for malting depends on many criteria, including the size, shape and uniformity of the kernels. Here, image analysis was used to measure kernel size and shape attributes (area, perimeter, length, width, F-circle and F-shape) in grain samples of 140 doubled-haploid lines from a two-rowed (cv Harrington) by six-rowed (cv Morex) barley cross. Interval mapping was used to map quantitative trait loci (QTLs) affecting the means and within-sample standard deviations of these attributes using a 107 marker genome map. Regions affecting one or more kernel size and shape traits were detected on all seven chromosomes. These included one near the vrs1 locus on chromosome 2 and one near the int-c locus on chromosome 4. Some, but not all, of the QTLs exhibited interactions with the environment and some QTLs affected the within-sample variability of kernel size and shape without affecting average kernel size and shape. When QTL analysis was conducted using data from only the two-rowed lines, the region on chromosome 2 was not detected but QTLs were detected elsewhere in the genome, including some that had not been detected in the analysis of the whole population. Analysis of only the six-rowed lines did not detect any QTLs affecting kernel size and shape attributes. QTL alleles that made kernels larger and/or rounder also tended to improve malt quality and QTL alleles that increased the variability of kernel size were associated with poor malt quality. PMID- 12582526 TI - Characterization of the main effects, epistatic effects and their environmental interactions of QTLs on the genetic basis of yield traits in rice. AB - Main effects, epistatic effects and their environmental interactions of QTLs are all important genetic components of quantitative traits. In this study, we analyzed the main effects, epistatic effects of the QTLs, and QTL by environment interactions (QEs) underlying four yield traits, using a population of 240 recombinant inbred lines from a cross between two rice varieties tested in replicated field trials. A genetic linkage map with 220 DNA marker loci was constructed. A mixed linear model approach was used to detect QTLs with main effects, QTLs involved in digenic interactions and QEs. In total, 29 QTLs of main effects, and 35 digenic interactions involving 58 loci were detected for the four traits. Thirteen QTLs with main effects showed QEs; no QE was detected for the QTLs involved in epistatic interactions. The amount of variations explained by the QTLs of main effect were larger than the QTLs involved in epistatic interactions, which in turn were larger than QEs for all four traits. This study illustrates the ability of the analysis to assess the genetic components underlying the quantitative traits, and demonstrates the relative importance of the various components as the genetic basis of yield traits in this population. PMID- 12582527 TI - Broccoli plants with pyramided cry1Ac and cry1C Bt genes control diamondback moths resistant to Cry1A and Cry1C proteins. AB - This study was undertaken to determine the effects of pyramiding two Bacillus thuringiensis (Bt) genes in the same plant on the production of Bt proteins and the control of diamondback moths (DBM, Plutella xylostella) resistant to one or the other protein. Broccoli lines carrying both cry1Ac and cry1C Bt genes were produced by sexual crosses of cry1Ac- and cry1C-transgenic plants. Plants containing both genes were selected by tests for resistance to kanamycin and hygromycin, and confirmed by PCR analysis for the Bt genes. Both cry1Ac and cry1C mRNAs were detected in the hybrid lines, and Cry1Ac and Cry1C proteins were stably produced at levels comparable to the parental plants. Plants producing both Cry1Ac and Cry1C proteins caused rapid and complete mortality of DBM larvae resistant to Cry1A or Cry1C, and suffered little or no leaf damage. These plants, in combination with the resistant DBM populations available, will allow greenhouse or field studies of resistance management strategies involving gene pyramiding. PMID- 12582528 TI - Development of molecular markers using MFLP linked to a gene conferring resistance to Diaporthe toxica in narrow-leafed lupin ( Lupinus angustifolius L.). AB - Phomopsis stem blight (PSB) caused by Diaporthe toxica is a major disease in narrow-leafed lupin ( Lupinus angustifolius L.). The F(2) progeny and the parental plants from a cross between a breeding line 75A:258 (containing a single dominant resistance gene Phr1 against the disease) and a commercial cultivar Unicrop (susceptible to the disease) were used for development of molecular markers linked to the disease resistance gene. Two pairs of co-dominant DNA polymorphisms were detected using the microsatellite-anchored fragment length polymorphism (MFLP) technique. Both pairs of polymorphisms were isolated from the MFLP gels, re-amplified by PCR, sequenced, and converted into co-dominant, sequence-specific and PCR-based markers. Linkage analysis by MAPMAKER suggested that one marker (Ph258M2) was 5.7 centiMorgans (cM) from Phr1, and the other marker (Ph258M1) was 2.1 cM from Ph258M2 but further away from Phr1. These markers are suitable for marker-assisted selection (MAS) in lupin breeding. PMID- 12582529 TI - RFLP mapping of a Hordeum bulbosum gene highly expressed in pistils and its relationship to homoeologous loci in other Gramineae species. AB - A cDNA sequence (Hbc8-2) isolated from pistils of the self-incompatible species Hordeum bulbosum was analysed for expression pattern and genetic map location. Hbc8-2 was expressed just prior to anthesis in mature pistils, and expression was maintained at a high level throughout anthesis. The same expression pattern was found in self-incompatible rye ( Secale cereale), but no expression was detected in the self-compatible cereals wheat ( Triticum aestivum) or barley ( Hordeum vulgare) at comparable stages of development. However, three wheat expressed sequence tags from a pre-anthesis library had high homology to Hbc8-2. Southern blot analyses using Hbc8-2 as a probe detected hybridising bands in the genomes of various Gramineae species including rye, barley, bread wheat, wild wheat relatives ( Aegilops tauschii and Ae. speltoides), oats ( Avena fatua and A. strigosa), rice ( Oryza sativa) and maize ( Zea mays). This suggests that Hbc8-2 like sequences are present in many species but that high levels of expression may be associated with self-incompatibility. Hbc8-2 was mapped on the long arms of chromosome 2H(b) of H. bulbosum, 2R of rye, and 2B and 2D of wheat and was assigned to chromosome 2H of barley using wheat/barley addition lines. On a H. bulbosum genetic map, Xhbc8-2 was located between Xbcd266 and Xpsr87, while in rye and wheat it was located in a 13.2-cM interval between Xpsr331 and Xpsr932, consistent with previous comparative mapping studies of these species. Mapping in rye suggested that Hbc8-2 is probably proximal to the Z self-incompatibility locus which was previously shown to be tightly linked to Xbcd266. PMID- 12582530 TI - An AFLP and RFLP linkage map and quantitative trait locus (QTL) analysis of growth traits in Salix. AB - A genetic linkage map of Salix (2n = 38), composed of 325 AFLP and 38 RFLP markers has been constructed. The map was based on a population ( n = 87) derived from a cross between the male hybrid clone "Bjorn" ( Salix viminalis x Salix schwerinii) and the female clone "78183" ( S. viminalis). Three hundred fifty seven AFLPs corresponding to DNA polymorphisms heterozygous in one parent and null in the other were scored. A total of 87 RFLP probes, most (83) derived from the Populus genome, yielded 39 and 11 polymorphic loci segregating in a 1:1 and 1:2:1 ratio respectively. Two maps, one for each parent, were constructed according to the "two-way pseudo-testcross" mapping strategy. The S. viminalis x S. schwerinii map (2,404 cM) included 217 markers and formed 26 major linkage groups while S. viminalis (1,844 cM) consisted of 146 markers placed on 18 major groups. In addition, eight and 14 additional minor linkage groups composed of less than four markers (doubles and triplets) were obtained in the S. viminalis x S. schwerinii and the S. viminalis maps, respectively. Both maps provided 70-80% genome coverage with an average density of markers of 14 cM. To investigate possible homologies between the parental maps, 20 AFLPs and 11 RFLPs segregating in 3:1 or 1:2:1 ratios were included in the linkage analysis. Eight linkage groups homologous between the two maps were detected. The present genetic map was used to identify quantitative trait loci (QTLs) affecting growth-related traits. Eleven QTLs were identified; seven QTLs for height growth, one QTL for stem diameter, one QTL for the height: diameter ratio, one QTL for the number of vegetative buds during flowering time and one QTL for the number of shoots. The estimated magnitude of the QTL effect ranged from 14 to 22% of the total phenotypic variance. One QTL associated with height growth and one affecting the height: diameter ratio were overlapping in the same marker interval with the QTL affecting stem diameter. QTL stability over years was estimated for traits measured in multiple years. Generally, QTLs were only significant in a single year although two QTLs for height growth were close to reaching the significance level in 2 consecutive years. PMID- 12582531 TI - AFLP analysis of genetic relationships among papaya and its wild relatives (Caricaceae) from Ecuador. AB - The AFLP technique was used to assess the genetic relationships among the cultivated papaya ( Carica papaya L.) and related species native to Ecuador. Genetic distances based on AFLP data were estimated for 95 accessions belonging to three genera including C. papaya, at least eight Vasconcella species and two Jacaratia species. Cluster analysis using different methods and principal co ordinate analysis (PCO), based on the AFLP data from 496 polymorphic bands generated with five primer combinations, was performed. The resulted grouping of accessions of each species corresponds largely with their taxonomic classifications and were found to be consistent with other studies based on RAPD, isozyme and cpDNA data. The AFLP analysis supports the recent rehabilitation of the Vasconcella group as a genus; until recently Vasconcella was considered as a section within the genus Carica. Both cluster and PCO analysis clearly separated the species of the three genera and illustrated the large genetic distance between C. papaya accessions and the Vasconcella group. The specific clustering of the highly diverse group of Vasconcella x heilbornii accessions also suggests that these genotypes may be the result of bi-directional introgression events between Vasconcella stipulata and Vasconcella cundinamarcensis. PMID- 12582532 TI - Genetic diversity in apricot revealed by AFLP markers: species and cultivar comparisons. AB - The genetic diversity of apricot ( Prunus armeniaca; 2n = 16) was studied using AFLP markers. Forty seven apricot cultivars were selected from the following geographic regions: Europe, North America, North Africa, Turkey, Iran and China. Five EcoRI- MseI AFLP primer combinations revealed 416 legible bands, of which 379 were polymorphic markers. A similarity matrix was prepared using the simple matching coefficient of similarity. A UPGMA dendrogram demonstrated a gradient of decreasing genetic diversity of varieties from the former USSR to Southern Europe. This is coherent with the historical dissemination of apricot from its center of origin in Asia. The American cultivars were intermediate demonstrating a different genetic base than the European and/or Mediterranean cultivars. Euclidean distances from the first ten Factorial Component Analysis coordinate axes were used to generate a tree using the Ward algorithm. The results of these analyses were evaluated based on the known geographic origins and agronomic characteristics of the cultivars studied. Four cultivar groups were identified: Diversification, Geographically Adaptable, Continental Europe and Mediterranean Basin. To evaluate the relationship of the common apricot with some closely related species, one or two accessions of the following related species or sub species from within the section Armeniaca were included in the analysis: Prunus armeniaca var. ansu, Prunus mume, Prunus brigantiaca, Prunus dasycarpa, and Prunus holosericea. A Neighbour Joining dendrogram was made using the similarity matrix. The P. holosericea accession fell well within the cultivar group, thus supporting its classification as a variant of P. armeniaca. The P. armeniaca var. ansu accession was sister to the common apricot cluster with a bootstrap value of 96%. P. mume was farther removed. P. brigantiaca was the most-distant from the common apricots. P. dasycarpa was intermediate between P. brigantiaca and P. mume, in accord with its plum-apricot hybrid origin. The results have a direct application for the selection of new breeding progenitors. PMID- 12582533 TI - Species relationships in Fagopyrum revealed by PCR-based DNA fingerprinting. AB - Random amplified polymorphic DNA (RAPD) markers were used to distinguish between 28 different accessions belonging to 14 species and two sub-species of Fagopyrum. Of the 75 random 10-mer primers tested, only 19 generated robust, easily interpretable amplification products. A total of 364 bands were observed with an average of 19.15 bands per primer, of which 99.45% were polymorphic. Primer OPN 08 produced the maximum number of fragments and UBC-183 produced the minimum number of fragments. The data were utilized to elucidate genetic relationships among 14 species and two sub-species of Fagopyrum. Cluster analysis using the unweighted paired group method of arithmetic means (UPGMA) showed four main clusters, two each of the cymosum and urophyllum groups. The results showed that Fagopyrum tataricum is closer to its wild ancestor F. tataricum ssp. potanini Batalin, closely followed by Fagopyrum giganteum. Cultivated common buckwheat ( Fagopyrum esculentum) showed affinity with its putative wild ancestor F. esculentum ssp. ancestrale and the other closely related diploid species Fagopyrum homotropicum. In the urophyllum group, Fagopyrum macrocarpum and Fagopyrum pleioramosum formed one cluster, whereas Fagopyrum capillatum, Fagopyrum gracilipes and Fagopyrum gilessii clustered separately. Except for a few cases, our results correspond with previously reported studies on Fagopyrum using the isozyme, RFLP and RAPD methods. Species-diagnostic amplification products specific to some species in the cymosum and urophyllum groups were identified. Our results show that RAPDs can be successfully used to analyze species relationships in Fagopyrum and also for constructing linkage maps. PMID- 12582534 TI - Mapping QTLs for field resistance to the rice blast pathogen and evaluating their individual and combined utility in improved varieties. AB - Lines from a Lemont x Teqing recombinant inbred population were evaluated for dilatory resistance to rice blast disease using: (1) the Standard Evaluation System (SES) for rating leaf blast, (2) the percentage diseased leaf area (%DLA), and (3) the area under a disease progress curve (AUDPC). RFLP mapping using 175 well-distributed loci revealed nine QTLs, one each on chromosomes 1, 2, 3, 4, 6, 7 and 9, with two loci on chromosome 12. All nine putative QTLs were associated with AUDPC, six with both a %DLA and a SES rating. Teqing contributed the resistance allele for all these loci except for the one located on chromosome 4. Individual QTLs accounted for 5-32% of the observed phenotypic variation, and combined QTL models accounted for 43-53%. Three QTLs were located near three of the four major resistance genes previously identified in this population. The resistances of both Lemont and Teqing were attributable to a combination of both major genes capable of inducing hypersensitive reactions and minor genes causing less-distinctive phenotypic differences. Interactions were noted between QTLs and major genes. Our findings are in support of the strategy of pyramiding major genes and QTLs in carefully selected combinations to develop improved varieties with resistance to the blast fungus that is both broad in spectrum and durable. PMID- 12582535 TI - Genetic relationships and discrimination of ten influential Upland cotton varieties using RAPD markers. AB - Influential Upland cotton ( Gossypium hirsutum L.) varieties are those that have the higher genetic contributions to modern Upland cultivars than other germplasms. Our previous research has shown significant differences in general combining ability (GCA) effects for yield, yield components, and fiber properties among ten influential cotton varieties. In this study, we used random amplified polymorphic DNA (RAPD) data to evaluate DNA variation of these ten varieties. Of 86 random decamer primers screened for their capability of amplifying DNA via the polymerase chain reaction (PCR), 63 generated a total of 312 DNA fragments. Forty two bands were polymorphic, which showed a low percentage (13.5%) of DNA variation among these influential varieties. Genetic similarities among the ten varieties based on RAPD data were from 92.7% to 97.6%. All of the varieties were individually identified by variety specific markers in genetic fingerprinting. One primer, UBC-149, amplified a 1,430-bp DNA fragment that was absent in five varieties and present in the other five varieties. This RAPD marker had significant negative relationships with GCA-effect estimates for seed cotton yield, lint yield, number of bolls per plant and micronaire, and significant positive relationships with GCA effects for boll size and seed index. This finding, for the first time, identifies a DNA fragment in cotton that is a potential DNA marker linked to a yield gene(s) or a yield-related gene(s). PMID- 12582536 TI - Molecular dissection of complex traits in autopolyploids: mapping QTLs affecting sugar yield and related traits in sugarcane. AB - Mapping quantitative trait loci (QTLs) for sugar yield and related traits will provide essential information for sugarcane improvement through marker-assisted selection. Two sugarcane segregating populations derived from interspecific crosses between Saccharum offinarum and Saccharum spontaneum with 264 and 239 individuals, respectively, were evaluated in three replications each for field performance from 1994 to 1996 at Weslaco, Texas. These two populations were analyzed for a total of 735 DNA marker loci to seek QTLs for sugar yield, pol, stalk weight, stalk number, fiber content and ash content. Among the 102 significant associations found between these six traits and DNA markers, 61 could be located on sugarcane linkage maps, while the other 41 were associated with unlinked DNA markers. Fifty of the 61 mapped QTLs were clustered in 12 genomic regions of seven sugarcane homologous groups. Many cases in which QTLs from different genotypes mapped to corresponding locations suggested that at least some of the QTLs on the same cluster might be different allelic forms of the same genes. With a few exceptions that explained part of the transgressive segregation observed for particular traits, the allele effects of most QTLs were consistent with the parental phenotype from which the allele was derived. Plants with a high sugar yield possessed a large number of positive QTLs for sugar yield components and a minimal number of negative QTLs. This indicates the potential effectiveness of marker-assisted selection for sugar yield in sugarcane. PMID- 12582537 TI - High-resolution genetic analysis of the Sd-1 aphid resistance locus in Malus spp. AB - Aphids cause serious physical and economic damage to most major crops throughout the world through feeding damage, with consequent symptom development and virus transmission. The rosy leaf-curling aphid ( Dysaphis devecta Wlk.) is a pest of apple ( Malus spp.) which displays an exceptionally clear phenotype with respect to susceptible and resistant symptoms. The Sd-1 locus for resistance to D. devecta biotypes 1 and 2 is present in Cox's Orange Pippin and its progeny and had previously been mapped to the top of linkage group 7. Detailed fine mapping of the locus was initiated with AFLP bulked segregant analysis of both pedigree and segregating bulks, which identified three new marker loci. Preliminary marker order in the Sd-1 region was established through mapping in a family derived from Prima x Fiesta, with additional segregation analysis on a Fiesta x Golden Delicious family. Previous recombinant data was re-evaluated and corrected. Two co-segregating AFLP fragments were found to contain a common (GA)(23) repeat, from which a PCR-based simple sequence repeat (SSR) assay was developed. A high resolution map around the Sd-1 region was established by analysing a large meta population of Sd-1 recombinants using 759 additional individuals from different families. The Sd-1 gene has been located within a 1.3-cM interval flanked by the molecular markers SdSSRa and 2B12a and co-locates with the RFLP marker MC064. Allelism between Sd-1 and Sd-2 resistant sources was tested. Molecular markers tightly linked to Sd-1 were shown to be co-segregating with the Sd-2 locus, which indicated that Sd-1 and Sd-2 loci are at least tightly linked and, probably, allelic. PMID- 12582538 TI - Development of RGA-CAPS markers and genetic mapping of candidate genes for sugarcane mosaic virus resistance in maize. AB - Three previously published resistance gene analogues (RGAs), pic13, pic21 and pic19, were mapped in relation to sugarcane mosaic virus (SCMV) resistance genes ( Scmv1, Scmv2) in maize. We cloned these RGAs from six inbreds including three SCMV-resistant lines (D21, D32, FAP1360A) and three SCMV-susceptible lines (D145, D408, F7). Pairwise sequence alignments among the six inbreds revealed a frequency of one single nucleotide polymorphism (SNP) per 33 bp for the three RGAs, indicating a high degree of polymorphism and a high probability of success in converting RGAs into codominant cleaved amplified polymorphic sequence (CAPS) markers compared to other sequences. SNPs were used to develop CAPS markers for mapping of the three RGAs in relation to Scmv1 (chromosome 6) and Scmv2 (chromosome 3), and for pedigree analyses of resistant inbred lines. By genetic mapping pic21 was shown to be different from Scmv2, whereas pic19 and pic13 are still candidates for Scmv1 and Scmv2, respectively, due to genetic mapping and consistent restriction patterns of ancestral lines. PMID- 12582539 TI - A resistance gene analog useful for targeting disease resistance genes against different pathogens on group 1S chromosomes of barley, wheat and rye. AB - Comparative sequence analysis of the resistance gene analog (RGA) marker locus aACT/CAA (originally found to be tightly linked to the multiallelic barley Mla cluster) from genomes of barley, wheat and rye revealed a high level of relatedness among one another and showed high similarity to a various number of NBS-LRR disease resistance proteins. Using the sequence-specific polymerase chain reaction (PCR), RGA marker aACT/CAA was mapped on group 1S chromosomes of the Triticeae and was associated with disease resistance loci. In barley and rye, the marker showed linkage to orthologous powdery mildew resistance genes Mla1 and Pm17, respectively, while in wheat linkage with a QTL against fusarium head blight (FHB) disease was determined. The use of RGA clones for R gene mapping and their role in the expression of qualitative and quantitative resistance is discussed. PMID- 12582540 TI - Genetic relationships and diversity of commercially relevant Echinacea species. AB - The genus Echinacea is comprised of nine species, which are perennial herbs indigenous to North America and which have been traditionally used as medicinal plants for centuries. Three Echinacea species, E. angustifolia, E. purpurea, and E. pallida, are currently being traded internationally in the natural products market. Echinacea products constitute a significant portion of this growing, multi-billion dollar industry. The increasing popularity of Echinacea products has led to the expansion of wildcrafting and commercial cultivation to meet the growing demand for plant material. Echinacea is considered of value as a nonspecific immune stimulant, and claims of its efficacy have been tentatively supported by both laboratory and clinical studies. This study used random amplified polymorphic DNA (RAPD) markers to determine the genetic relationships of the three Echinacea species of commercial interest, to evaluate the level of diversity present within germplasm of each of the three species, and to compare accessions of each species available from different sources. A total of 101 RAPD markers were generated for the 76 individuals of four species included in the analysis. NTSYS-pc was used to evaluate the genetic relationships of the three species and to determine the general level of overall diversity. Analysis of molecular variance (AMOVA) was performed using pruned marker sets corrected for the dominant nature of RAPD markers. AMOVA revealed that most of the variation occurred within accessions of the same species, though some accessions of both E. pallida and E. angustifolia were found to be significantly different from other accessions of the same species. PMID- 12582541 TI - Genetic control of monoterpene composition in the essential oil of Melaleuca alternifolia (Cheel). AB - Previous studies into the population structure of Melaleuca alternifolia by both isozyme and microsatellite analysis revealed little evidence for genetic structuring within genetic provenances. In contrast, analysis of the oil composition within these same regions showed distinct clustering of chemotypes within the provenances suggesting either that chemotype was not under genetic control, or that there is strong environmental selection for plant chemotypes. To investigate the level of genetic control of monoterpene composition in the essential oil of M. alternifolia, individuals representing the three extreme chemotypes of high terpinen-4-ol, high 1,8-cineole and high terpinolene were crossed with an individual with the commercially desirable high terpinen-4-ol oil profile. The progeny resulting from these crosses displayed oil profiles that were intermediate to that of the parent. Further analysis of the survey of oil chemotypes within the natural population also suggests that these intermediate chemotypes may arise naturally between regions containing high proportions of the extreme chemotypes. These results imply that there is a level of genetic structure for chemotype determination within the genetic provenance that is undetected by isozyme and microsatellite analysis. This information could play a vital role in the selection of appropriate genetic material to be used in future essential oil selection and breeding programs. PMID- 12582542 TI - Lactuca saligna, a non-host for lettuce downy mildew ( Bremia lactucae), harbors a new race-specific Dm gene and three QTLs for resistance. AB - Lactuca sativa (lettuce) is susceptible to Bremia lactucae (downy mildew). In cultivated and wild Lactuca species, Dm genes have been identified that confer race-specific resistance. However, these genes were soon rendered ineffective by adaptation of the pathogen. Lactuca saligna (wild lettuce) is resistant to all downy mildew races and can be considered as a non-host. Therefore, L. saligna might be an alternative source for a more-durable resistance to downy mildew in lettuce. In order to analyze this resistance, we have developed an F(2) population based on a resistant L. saligna x susceptible L. sativa cross. This F(2) population was fingerprinted with AFLP markers and tested for resistance to two Bremia races NL14 and NL16. The F(2) population showed a wide and continuous range of resistance levels from completely resistant to completely susceptible. By comparison of disease tests, we observed a quantitative resistance against both Bremia races as well as a race-specific resistance to Bremia race NL16 and not to NL14. QTL mapping revealed a qualitative gene ( R39) involved in the race specific resistance and three QTLs ( RBQ1, RBQ2 and RBQ3) involved in the quantitative resistance. The qualitative gene R39 is a dominant gene that gives nearly complete resistance to race NL16 in L. saligna CGN 5271 and therefore it showed features similar to Dm genes. The three QTLs explained 51% of the quantitative resistance against NL14, which indicated that probably only the major QTLs have been detected in this F(2) population. The perspectives for breeding for durable resistance are discussed. PMID- 12582543 TI - Pitfalls of genetic analysis using a doubled-haploid backcrossed to its parent. AB - A population derived from a doubled-haploid backcrossed to its parent has a unique genetic structure, which affects the mapping and genetic analysis of molecular-marker data. In such a cross: (1) repulsion linkages are reduced dramatically and can be detected only in restricted chromosome sections; each repulsion-linkage group represents a section of a chromosome carrying a crossover that occurred in the meiosis that produced the gamete from which the doubled haploid was derived; (2) the number of coupling-linkage groups observed depends on how many crossovers occurred during the meiosis; the observed size of a linkage group will be only a fraction of a total chromosome if the chromosome carried in the doubled-haploid resulted from crossover exchanges; (3) the size covered by all the observed linkages is only equivalent to the haploid genome; and (4) the ratio of single-dose to multi-dose markers is inflated. These features have not been recognized in previous reports, resulting in misinterpretations in genetic mapping and analysis of the molecular-marker data. The ratio of single- to multi-dose markers has been used inappropriately to distinguish polyploid type. PMID- 12582544 TI - Genetics of ethylene biosynthesis and restriction fragment length polymorphisms (RFLPs) of ACC oxidase and synthase genes in melon ( Cucumis melo L.). AB - We investigated the genetics of ethylene biosynthesis and its linkage to the RFLPs of the ACC oxidase and synthase genes in melon ( Cucumis melo L.). The results suggested that the A(0) and B(0) fragments of RFLP-MEL1 of the ACC oxidase gene were two alleles from a single locus, as were the B and C fragments of RFLP-MEACS1 of the ACC synthase gene. The B(0) allele seemed to be partially dominant over the A(0) allele, whereas B and C alleles appeared to map to quantitative trait loci (QTLs), which most likely contributed to ethylene production. Both RFLPs were linked to ethylene production rates, but they were not linked to each other. The interaction effects of the ACC oxidase and synthase genes on ethylene production were revealed by segregation of RFLP-MEL1 and RFLP MEACS1. The results of single-copy-reconstruction assays suggested that the ACC oxidase gene is a single copy, whereas the ACC synthase gene is a component of a multigene family in the melon genome. The abscission phenotype appeared to be controlled by an independent locus, with the abscission (full-slip) allele dominant over the non-abscission (not full-slip) allele. These results may facilitate efforts toward mapping the quantitative trait loci (QTLs) of ethylene production. The RFLPs may be used in marker-assisted selection in developing melons with a more-desirable low ethylene production rate for enhancing postharvest storage life. PMID- 12582545 TI - Assessment of the C(4) phosphoenolpyruvate carboxylase gene diversity in grasses (Poaceae). AB - C(4) phosphoenolpyruvate carboxylase (PEPC) is a key enzyme in the C(4) photosynthetic pathway. To analyze the diversity of the corresponding gene in grasses, we designed PCR primers to specifically amplify C(4) PEPC cDNA fragments. Using RT-PCR, we generated partial PEPC cDNA sequences in several grasses displaying a C(4) photosynthetic pathway. All these sequences displayed a high homology (78-99%) with known grass C(4) PEPCs. PCR amplification did not occur in two grasses that display the C(3) photosynthetic pathway, and therefore we assumed that all generated sequences corresponded to C(4) PEPC transcripts. Based on one large cDNA segment, phylogenetic reconstruction enabled us to assess the relationships between 22 grass species belonging to the subfamilies Panicoideae, Arundinoideae and Chloridoideae. The phylogenetic relationships between species deduced from C(4) PEPC sequences were similar to those deduced from other molecular data. The sequence evolution of the C(4) PEPC isoform was faster than in the other PEPC isoforms. Finally, the utility of the C(4) PEPC gene phylogeny to study the evolution of C(4) photosynthesis in grasses is discussed. PMID- 12582546 TI - Genetic mapping of 66 new microsatellite (SSR) loci in bread wheat. AB - In hexaploid bread wheat ( Triticum aestivum L. em. Thell), ten members of the IWMMN ( International Wheat Microsatellites Mapping Network) collaborated in extending the microsatellite (SSR = simple sequence repeat) genetic map. Among a much larger number of microsatellite primer pairs developed as a part of the WMC ( Wheat Microsatellite Consortium), 58 out of 176 primer pairs tested were found to be polymorphic between the parents of the ITMI ( International Triticeae Mapping Initiative) mapping population W7984 x Opata 85 (ITMI pop). This population was used earlier for the construction of RFLP ( Restriction Fragment Length Polymorphism) maps in bread wheat (ITMI map). Using the ITMI pop and a framework map (having 266 anchor markers) prepared for this purpose, a total of 66 microsatellite loci were mapped, which were distributed on 20 of the 21 chromosomes (no marker on chromosome 6D). These 66 mapped microsatellite (SSR) loci add to the existing 384 microsatellite loci earlier mapped in bread wheat. PMID- 12582547 TI - Structure and expression of the Zea mays mutS-homologs Mus1 and Mus2. AB - DNA mismatch repair proteins play an important role in maintaining the integrity of the genetic information during replication and homologous recombination. The MutS-homologous (MSH) and MutL-homologous (MLH) proteins are highly conserved among all prokaryotes and eukaryotes. We have isolated two mutS homologous genes from Zea mays, named Mus1 and Mus2. Phylogenetic analysis identifies Mus1 as a member of the MSH2 protein family. Mus2 is an ortholog of the Arabidopsis thaliana MSH7 protein and belongs to a subgroup of MSH proteins that is possibly plant-specific. Mus1 and Mus2 are expressed at very low levels. Mus1 is located on chromosome 7L near locus b32B, and mus2 maps on chromosome 3S. PMID- 12582548 TI - AFLP markers demonstrate local genetic differentiation between two indigenous oak species [ Quercus robur L. and Quercus petraea (Matt.) Liebl.] in Flemish populations. AB - The nuclear genetic variation within and between four sessile ( Q. petraea) and six pedunculate ( Q. robur) autochthonous Flemish oak populations was investigated with AFLP markers. One sessile and one pedunculate oak population were additionally screened for detailed leaf characteristics using an image analysis system. Principal coordinate analysis on the AFLP data classified the oaks in two main groups, according to their taxonomic status. No species-specific AFLP markers were found using four primer combinations, but marker frequency differences up to 71% were recorded between both species. Analysis of the genetic structure showed that the divergence between species, as observed by ordination, was significant. Both species revealed similar diversity levels. A smaller though significant differentiation was also revealed for both species among populations within species. Molecular and morphology based approaches showed a high degree of consistency. Screening of 60 AFLP primer combinations using a bulking strategy did not allow identifying species-specific markers, which supports the conclusions reached in previous studies. The distribution of genetic variability at the species and at the population level is discussed. PMID- 12582549 TI - Improvement of hybrid yield by advanced backcross QTL analysis in elite maize. AB - We applied an advanced backcross breeding strategy to identify quantitative trait loci (QTLs) of agronomic importance in a cross between two elite inbreds of maize, RD6502 (Mo17-type recurrent parent) and RD3013 (Iodent donor parent). Two hundred and four BC(2) families were scored at 106 SSR, 15 AFLP, and 38 Heartbreaker (MITE) loci. BC(2) testcrosses (TC) with B73 were phenotyped at six locations in the Midwest and N.Y. We detected four grain yield, six grain moisture, and three plant height QTLs at which the RD3013 allele had a favorable effect ( p < 0.05). All four yield QTLs were selected as target introgressions in the development of BC(3)TC families. As predicted by BC(2)TC analysis, BC(3)TC entries containing introgressions at yld3.1 and yld10.1 significantly outperformed non-carrier entries by 11.1% (15.6 bu/A at one location) and 6.7% (7.1 bu/A averaged across two locations), respectively, in replicated Midwestern trials ( p < 0.05). Detection of yld10.1 effects in the BC(2)TC by spatial analysis (i.e., incomplete block, response surface, autoregressive, moving average or autoregressive moving average), but not by conventional single point analysis or interval mapping, indicated the utility of local environmental control for QTL mapping in unreplicated maize progeny. This work demonstrated that the advanced backcross QTL method can be applied to identify and manipulate useful QTLs in heterotic inbreds of elite maize. Genetic gains by this approach can be coupled with the maintenance and selection of favorable epistatic gene complexes by traditional hybrid breeding for maize improvement. PMID- 12582550 TI - Fluorescent-labeled microsatellite panels useful for detecting allelic diversity in cultivated rice ( Oryza sativa L.). AB - Four multiplex panels of fluorescent-labeled rice microsatellite markers were designed to survey allelic diversity at a total of 27 simple sequence repeat loci. Automated fragment detection, size calling and binning were used to identify the microsatellite alleles. The panels were tested on 72 accessions representing the diversity of ecotypes and isozyme groups in cultivated rice ( Oryza sativa L.). Genetic diversity was evaluated by estimating the distribution and range of allele sizes, as well as the polymorphism information content (PIC), for each microsatellite locus. The multiplex panels were shown to be useful for fingerprinting and for clustering rice varieties, and genetic associations determined in this analysis agreed well with previous isozyme- and RFLP-based studies of these genotypes. Because of the wide cross-section of germplasm represented, we expect that the allele size-ranges reported here are close to the maximum for cultivated rice. In the future, panels of microsatellites from each rice chromosome will be made to facilitate the mapping of segregating populations and the identification of genes and QTLs underlying traits of interest. PMID- 12582551 TI - Identification of AFLP and microsatellite markers linked with an aluminium tolerance gene in barley ( Hordeum vulgare L.). AB - Barley is the most sensitive among the cereals to aluminium (Al) stress and breeding for more tolerant cultivars is a priority. To enhance selection efficiency for Al tolerance in barley, PCR-based AFLP and microsatellite markers linked to a locus conferring tolerance to aluminium were identified. The study used F(2) progeny derived from a single cross between Yambla (moderately tolerant of Al) and WB229 (tolerant of Al) and developed hydroponic pulse-recovery screening methods to assess tolerance of phenotypes based on root growth. The segregation ratios of tolerant and sensitive genotypes and F(3) progeny testing suggest that a single major gene controlled Al tolerance ( Alt). In order to determine the chromosomal location of the Alt gene, we used the AFLP technique coupled with bulk segregant analysis. We evaluated tolerant and sensitive bulks using 30 combinations of EcoRI/ MseI primers, and 12 of these permitted differentiation of the sensitive and tolerant bulks. More than 1,000 amplified fragments were obtained, and 98 polymorphic bands were scored. AFLP analysis of wheat-barley chromosome addition lines indicated that the Alt gene was located on barley chromosome 4H. Four chromosome 4H-specific microsatellite markers (Bmac310, Bmag353, HVM68 and HVMCABG) were tightly linked to Alt. The large allelic variation detected with microsatellite marker Bmag353 allowed us to implement this marker for routine marker-assisted selection for Al tolerance, and 396 plants could be screened on a single gel. PMID- 12582552 TI - Genotype-environment interactions in pollen competitive ability in an anemophilous tree, Betula pendula Roth. AB - This study describes genotype-environment interactions in pollen competitive ability expressed as pollen-tube growth rate and seed-siring success in Betula pendula Roth. A factorial crossing design was applied using the same maternal and paternal clones in two different environmental conditions, in a B. pendula seed orchard established in a greenhouse and at an outdoor clone collection. Both single donor and two-donor pollinations were employed. Female inflorescences were collected after a fixed time of germination, pollen-tube lengths were measured for each cross, and paternity of the seeds sired by two-donor pollen mixtures was analyzed using isozyme markers. The pollination site had a significant influence on pollen-tube growth rate and seed-siring success. Significant interactions between pollination site and pollen donor indicated genotype-environment interactions in pollen-tube growth rate and seed-siring success. A highly significant positive correlation between pollen-tube growth rate and seed-siring success was found in the greenhouse but not at the outdoor clone collection. These results suggest that the pollen-tube growth rate can be a predictor of seed siring success in controlled greenhouse conditions, where differences among maternal plants are mainly of genetic origin, but not in more heterogeneous outdoor conditions. In natural birch stands, environmental maternal effects probably diminish the significance of pollen competition for sexual selection in Betula pendula. At seed orchards, the effects of environmental conditions on pollen competitive ability can have important consequences for the genetic composition of the seed crop. PMID- 12582553 TI - Conservation and synteny of SSR loci and QTLs for vegetative propagation in four Eucalyptus species. AB - Conservation of microsatellite loci, heterozygous in Eucalyptus grandis, Eucalyptus urophylla, Eucalyptus tereticornis and Eucalyptus globulus, allowed us to propose homeologies among genetic linkage groups in these species, supported by at least three SSR loci in two different linkage groups. Marker-trait associations for sprouting and adventitious rooting ability were also compared in the four species. Putative quantitative trait loci (QTLs) influencing vegetative propagation traits were located on homeologous linkage groups. Our findings indicate high transferability of microsatellite markers between Eucalyptus species of the Symphyomyrtus subgenus and establish foundations for the use of synteny in the genetic analysis of this genus. Microsatellite markers should help integrate eucalypt genetic linkage maps from various sources. The availability of comparative linkage maps provides a basis of more-efficient use of genetic information for molecular breeding and evolutionary studies in Eucalyptus. PMID- 12582554 TI - Resistance gene analogues of chickpea ( Cicer arietinum L.): isolation, genetic mapping and association with a Fusarium resistance gene cluster. AB - Resistance gene analogues (RGAs) of Cicer were isolated by different PCR approaches and mapped in an inter-specific cross segregating for fusarium wilt by RFLP and CAPS analysis. Initially, two pairs of degenerate primers targeting sequences encoded at nucleotide-binding sites (NBS), which are conserved in plant disease resistance genes such as RPS2, L6 and N, were selected for amplification. Cloning and sequence analysis of amplified products from C. arietinum DNA revealed eight different RGAs. Additionally, five RGAs were identified after characterisation of the presumptive RGA alleles from C. reticulatum. Therefore, a total of 13 different RGAs were isolated from Cicer and classified through pair wise comparison into nine distinct classes with sequence similarities below a 68% amino acid identity threshold. Sequence comparison of seven RGA alleles of C. arietinum and C. reticulatum revealed polymorphisms in four RGAs with identical numbers of synonymous and non-synonymous substitutions. An NlaIII site, unique in the RGA-A allele of C. arietinum, was exploited for CAPS analysis. Genomic organisation and map position of the NBS-LRR candidate resistance genes was probed by RFLP analysis. Both single-copy as well as multi-copy sequence families were present for the selected RGAs, which represented eight different classes. Five RGAs were mapped in an inter-specific population segregating for three race specific Fusarium resistances. All RGAs mapped to four of the previously established eight linkage groups for chickpea. Two NBS-LRR clusters were identified that could not be resolved in our mapping population. One of these clusters, which is characterised by RFLP probe CaRGA-D, mapped to the linkage group harbouring two of three Fusarium resistance genes characterised in the inter-specific population. Our study provides a starting point for the characterisation and genetic mapping of candidate resistance genes in Cicer that is useful for marker-assisted selection and as a pool for resistance genes of Cicer. PMID- 12582555 TI - Mixing of maize and wheat genomic DNA by somatic hybridization in regenerated sterile maize plants. AB - Intergeneric somatic hybridization was performed between albino maize ( Zea mays L.) protoplasts and mesophyll protoplasts of wheat ( Triticum aestivum L.) by polyethylene glycol (PEG) treatments. None of the parental protoplasts were able to produce green plants without fusion. The maize cells regenerated only rudimentary albino plantlets of limited viability, and the wheat mesophyll protoplasts were unable to divide. PEG-mediated fusion treatments resulted in hybrid cells with mixed cytoplasm. Six months after fusion green embryogenic calli were selected as putative hybrids. The first-regenerates were discovered as aborted embryos. Regeneration of intact, green, maize-like plants needed 6 months of further subcultures on hormone-free medium. These plants were sterile, although had both male and female flowers. The cytological analysis of cells from callus tissues and root tips revealed 56 chromosomes, but intact wheat chromosomes were not observed. Using total DNA from hybrid plants, three RAPD primer combinations produced bands resembling the wheat profile. Genomic in situ hybridization (GISH) using total wheat DNA as a probe revealed the presence of wheat DNA islands in the maize chromosomal background. The increased viability and the restored green color were the most-significant new traits as compared to the original maize parent. Other intermediate morphological traits of plants with hybrid origin were not found. PMID- 12582556 TI - Mapping of DNA markers to arms and sub-arm regions of Nicotiana sylvestris chromosomes using aberrant alien addition lines. AB - Seven monosomic addition plants, each containing the full complement of Nicotiana plumbaginifolia (2n = 20, genome constitution PP) and an aberrant chromosome of Nicotiana sylvestris (2n = 24, SS), were produced from backcrosses of hyperdiploid derivatives of the sesquidiploid hybrid PPS to N. plumbaginifolia. The N. sylvestris chromosomes in these plants were characterized by karyotype analysis, Southern hybridization with DNA markers previously localized on N. sylvestris chromosomes and a 269-bp fragment from the 3' end of 25S rDNA, and fluorescence in situ hybridization using 25S rDNA, 5S rDNA and telomere repeats (TTTAGGG)(n) as probes. The N. sylvestris chromosomes in these plants were identified to be telocentrics 6S, 7S and 8S, and deletions 7S, 10, 12S and 12L, respectively. The successful identification of aberrant chromosomes in these lines enabled us to assign DNA markers to arms and sub-arm regions of N. sylvestris chromosomes. All aberrant chromosomes in the addition lines could be transmitted through mitosis and meiosis. The potential applications of the addition lines in high-resolution physical mapping, the isolation of N. sylvestris chromosomes by flow cytometry, and an understanding of the chromosomal distribution of 45S rDNA in N. sylvestris are discussed. PMID- 12582557 TI - A bentazon and sulfonylurea sensitive mutant: breeding, genetics and potential application in seed production of hybrid rice. AB - The use of a thermosensitive genic male sterility (TGMS) system in two-line hybrid rice breeding is affected greatly by the sterility instability of TGMS lines caused by temperature fluctuation beyond their critical temperatures for fertility reversion. To prevent seed production from self contamination, we have developed a system to secure seed purity using a herbicide-sensitive TGMS mutant, M8077S, obtained by radiation. Genetic analysis, using the F(1), F(2) and F(3) populations derived from this mutant and other normal varieties, revealed that bentazon lethality/sensitivity was controlled by a single recessive gene, which was named bel. The mutant can be killed at the seedling stage by bentazon at 300 mg/l or higher, a dosage that is safe for its F(1) hybrids and all other normal varieties. This mutant is also sensitive to all the tested sulfonylurea herbicides. Response of segregating plants to these two types of herbicide indicated that sulfonylurea sensitivity was also controlled by bel. By crossing this mutant with Pei-Ai 64S, an F(2) population was developed for genetic mapping. Surveying the two DNA pools from sensitive and non-sensitive F(2) plants identified four markers that were polymorphic between the pools. The putative linked markers were then confirmed with the F(2) population. The bel locus was located on chromosome 3, 7.1 cM from the closest microsatellite marker RM168. Phenotypic analysis indicated that the bel gene had no negative effect on agronomic traits in either a homozygous or heterozygous status. The mutant M8077S is valuable in the development of a TGMS breeding system for preventing impurity resulting from temperature fluctuation of the TGMS. Several two-line hybrid rice crosses using this system are under development. PMID- 12582558 TI - Tissue-culture enhanced transposition of the maize transposable element Dissociation in Brassica oleracea var. ' Italica' AB - To investigate the potential of heterologous transposons as a gene tagging system in broccoli ( Brassica oleracea var. Italica), we have introduced a Ds-based two element transposon system. Ds has been cloned into a 35S-SPT excision-marker system, with transposition being driven by an independent 35S-transposase gene construct ( Tpase). In three successive selfed generations of plants there was no evidence of germinal-excision events. To overcome this apparent inability to produce B. oleracea plants with germinal excisions, we performed a novel tissue culture technique to select for fully green shoots from seed with somatic excision events. The results showed a very high efficiency of regeneration of fully green plants (up to 65%) and molecular analysis indicated that the plants genetically were like plants that contain a germinal-excision event. Further molecular analysis of these plants showed that 69% exhibited reinsertion of Ds back into the plant genome. Sequencing of donor-site footprints after Ds excision, revealed that there is an indication of more-severe deletions and rearrangements when higher concentrations of streptomycin are used in the tissue culture selection process. Adapted versions of this regeneration technique have a high potential for providing germinal excision-like events in heterologous plants species which show low transposon activity. Alternatively, there is the potential to increase the proportion of 'germinal' plants in earlier generations of more active plant species. PMID- 12582559 TI - Molecular cloning, characterization, expression and chromosomal location of OsGAPDH, a submergence responsive gene in rice ( Oryza sativa L.). AB - Differential clones from submergence stress and control treatment from rice seedlings were isolated by the differential screening method. One of the clones, OsGAPDH, represented a gene that was expressed at high level during 12-h submergence. A homology search of GenBank databases showed that OsGAPDH had significant sequence homology with maize non-reversible glyceraldehyde-3 Phosphate dehydrogenase. The OsGAPDH sequence consists of 1,772 bp with the longest open reading frame encoding 499 amino acids with a calculated relative mass of 54.2 kDa. Genomic Southern analysis indicated that one or two copies of the OsGAPDH gene occur in the Yukihikari genome. The chromosomal location of the OsGAPDH gene was identified by RFLP analysis indicating that OsGAPDH was located on chromosome 8. Tissue-specific expression of OsGAPDH indicated that the high level of mRNA was detected in the panicle. Plants exposed to drought, submergence and ABA treatment showed an increased accumulation of OsGAPDH transcripts. The induction of Escherichia coli cells containing the pGST-OsGAPDH plasmid resulted in the accumulation of a large amount of the 83.2-kDa recombinant protein. The purified GAPDH enzyme showed an optimum activity at pH 8.5 and 50 degrees C, and was strongly inhibited by ATP and ADP. PMID- 12582560 TI - Identification of novel low M(r) glutenin subunits in the high quality bread wheat cv Salmone and their effects on gluten quality. AB - Southern-blot hybridization with a probe specific for genes encoding for low M(r) glutenin subunits showed that the high quality bread wheat cv Salmone contains two DNA fragments designated as SF720 and SF750. These fragments were found to occur on the chromosome-1B satellite, and to be associated with the presence of two strongly staining low M(r) glutenin subunits in the two-dimensional A-PAGE x SDS-PAGE pattern of cv Salmone. Comparison of 65 F(6) random lines derived from the cross between cv Salmone and the medium quality line FAP74809 revealed that the presence of fragments SF720 and SF750 had significant positive effects on several quality related parameters such as SDS sedimentation volume, Farinograph stability and Alveograph strength (W), tenacity (P) and elasticity (L). Additive effects of high M(r) glutenin subunits 1 and 7+9 on gluten quality were found as well. Fragments SF720 and SF750 were suggested to occur at a locus other than Glu B3, as indicated by their relatively high frequency of recombination with the Gli B1 locus. PMID- 12582561 TI - PCR Sampling of disease resistance-like sequences from a disease resistance gene cluster in soybean. AB - Clusters of Resistance-like genes (RLGs) have been identified from a variety of plant species. In soybean, RLG-specific primers and BAC-fingerprinting were used to develop a contig of overlapping BACs for a cluster of RLGs on soybean linkage group J. The resistance genes Rps2 ( Phytophthora stem and root rot) and Rmd-c (powdery mildew) and the ineffective nodulation gene Rj2 were previously mapped to this region of linkage group J. PCR hybridization was used to place two TIR/NBD/LRR cDNAs on overlapping BACs from this contig. Both of the cDNAs were present on BAC 34P7. Fingerprinting of this BAC suggested as many as twelve different RLGs were present. Given the high nucleotide identity shared between cDNAs LM6 and MG13 (>90%), direct sequencing of this region would be difficult. More sequence information was needed about the RLGs within this region before sequencing could be undertaken. By comparing the genomic sequences of cDNAs LM6 and MG13 we identified conserved regions from which oligonucleotide primers specific to BAC 34P7 RLGs could be designed. The nine primer pairs spanned the genomic sequence of LM6 and produced overlapping RLG products upon amplification of BAC 34P7. Amplification products from 12 different RLGs were identified. On average, nucleotide identity between RLG sequences was greater than 95%. Examination of RLG sequences also revealed evidence of additions, deletions and duplications within targeted regions of these genes. Using previously mapped cDNAs we were able to quickly and inexpensively access multiple RLGs within a single specific cluster. PMID- 12582562 TI - AFLP diversity in the common vetch ( Vicia sativa L.) on the world scale. AB - The Vavilov Institute of Plant Industry (VIR) keeps a living seed collection of about 700 accessions of landraces and local cultivars of common vetch ( Vicia sativa L.) that have been collected over a period of more than 50 years throughout the former USSR. Much of the material is available nowhere else. The collection of this economically important fodder crop is well adapted to the various growing regions of Russia and serves as a basis for the all domestic vetch breeding programs. Using AFLP as a DNA fingerprinting method we investigated 673 accessions from the VIR and compared their genetic variability with that of the worldwide vetch collection of the Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), 450 accessions. The analysis is a first assessment of the intra-specific diversity of V. sativa stored ex situ on a scale of more than 1,000 accessions. Six primer combinations, which gave clear polymorphic amplification products with 96 test samples, were chosen from 111 primer combinations tested. The selected AFLP primers used to analyse the V. sativa intra-specific diversity resulted in 70 unequivocally recognizable polymorphic fragments. We found that all of the AFLP fragments generated can be detected with varying frequency throughout the entire distribution area of V. sativa. The difference in frequency of some AFLP fragments between the regions may amount to 90%. The arrangement of most of the accessions in all dendrograms reflects their geographical origin, with a differentiation between Russia, Western Europe, Turkey and Bulgaria, and the Mediterranean. The "Russian" genepool stored at the IPK is a limited and biased sample of the available diversity when compared to the material stored at the VIR. Approximately 10-15% of the accessions in each geographical group showed AFLP patterns that clustered with members of other groups. This appreciable overlap raises several questions: (1) to which degree is an AFLP pattern representative of the overall genetic similarity of the samples; (2) to which degree are samples collected at a site adaptively limited to that site? Since our data identify accessions with very similar AFLP patterns from very diverse geographic origins, a comparison of the agronomic performance of these accessions (possibly in the two regions) will provide important information for the utilization of ex situ germplasm collections. PMID- 12582563 TI - Mapping QTLs for resistance to the cyst nematode Globodera pallida derived from the wild potato species Solanum vernei. AB - Resistance to the potato cyst nematode (PCN) species Globodera pallida, derived from the wild diploid potato species Solanum vernei, has been investigated. This source of resistance, which is effective against all of the major pathotypes of G. pallida and Globodera rostochiensis, has been assumed to be due to several genetic factors, but it has proved difficult to deploy effectively in breeding strategies for potato cultivars. Diploid and tetraploid potato populations segregating for 'vernei' resistance were analysed. At the tetraploid level, a bulk segregant analysis (BSA) approach was employed and detected AFLP markers linked to a resistance QTL on potato linkage group V. Conventional linkage analysis of a diploid population identified QTL on linkage groups V and IX. A marker linked to a QTL on linkage group V has been converted to a single-locus PCR-based marker, which can be used to detect the presence of the QTL in diploid and tetraploid potato germplasm. Moreover, there is evidence that one of the AFLPs detected by BSA appears to be specific to an introgressed segment of DNA from S. vernei. These results are compared with those obtained from other studies on resistance to the PCN species G. pallida. PMID- 12582564 TI - Destiny of a transgene escape from Brassica napus into Brassica rapa. AB - Transgenic Brassica napus can be easily crossed with wild Brassica rapa. The spread of the transgene to wild species has aroused the general concern about its effect on ecological and agricultural systems. This paper was designated, by means of population genetics, to study the fate of a transgene escape from B. napus to B. rapa. Three models were proposed to survey the change in gene frequency during successive backcross processes by considering selection pressures against aneuploids, against herbicide-susceptible individuals, and by considering A-C intergenomic recombination and the effect of genetic drift. The transmission rate of an A-chromosome gene through an individual to the next generation was 50%, irrespective of the chromosome number; while that of a C chromosome transgene varied from 8.7% to 39.9%, depending on the chromosome number of the individual used in the backcross. Without spraying herbicide, the frequency of an A-chromosome gene was 50% in the BC(1) generation, and decreased by 50% with the advance of each backcross generation; that of a C-chromosome gene was around 39.9% in BC(1), 7.7% in BC(2), 1.2% in BC(3) and 0.1% in the BC(4) generation. Under the selection pressure against herbicide-susceptible individuals, the frequency of a transgene reached a stable value of about 5.5% within six generations of successive backcrossings. The effect of genetic drift and intergenomic exchange on gene transmission rate was discussed. It is suggested that the transgene integrated on a C-chromosome (or better on a cytoplasm genome) is safer than that on an A-chromosome. The transgenic cultivars should be cultivated rotationally by year(s) with other non-transgenic varieties in order to reduce the transfer of the transgene to wild B. rapa species. PMID- 12582565 TI - Molecular mapping of the intermedium spike-c ( int-c) and non-brittle rachis 1 ( btr1) loci in barley ( Hordeum vulgare L.). AB - Lateral spikelet fertility and a non-brittle rachis are key characters in studying the evolution of barley. The fertility of lateral spikelets is controlled predominantly by the alleles at the vrs1 locus on chromosome 2HL and is modified by the alleles at the int-c locus on chromosome 4HS. The non-brittle rachis is controlled by alleles at two tightly linked loci, btr1 and btr2, on chromosome 3HS. This paper presents the integration of the int-c and btr1 loci in molecular linkage maps of barley. The int-c locus was mapped to the end of chromosome 4HS, 8.2 cM distal from the MWG2033 locus. The analysis was followed by a composite interval mapping of quantitative trait loci, which verified the position of the int-c locus. Linkage analysis using recombinant inbred lines showed that the btr1 locus is flanked between two AFLP loci, e14m27.4.1 and e15m19.7, with map distances of 3.1 cM and 4.2 cM, respectively. The molecular markers will expedite further high-density mapping of the int-c and btr1 loci. PMID- 12582566 TI - RFLP diversity and relationships among traditional European maize populations. AB - Given the large extent of hybrid cultivation, the importance of conserving the diversity of crop genetic resources has given birth to numerous collections of old races. In the present paper, we conduct a molecular characterisation of a large collection of 488 European maize populations using the bulk RFLP analysis. The analysis of 23 RFLP loci showed a high allelic richness of 11.5 alleles per locus. Populations from eastern Europe (Poland, Austria, Germany, etc.) showed the lowest genetic diversity, a lower number of unique alleles and a higher percentage of fixed loci than populations from southern Europe. In fact, genetic diversity appeared higher in Southern regions where the first maize populations are thought to have been introduced. Molecular classification based on Rogers' distance (i.e. alleles frequencies) allowed us to distinguish three main clusters which were highly consistent with geographic origins. A Northeastern cluster grouped together early or intermediate populations from Northeastern countries and the Balkans, a southeastern cluster joined late and partially dent populations from Greece and Italy, and, a southwestern cluster was made up of early flint populations from northern Spain, Portugal and the Pyrenees. A correlation between allelic frequencies at some loci and latitude and/or longitude was observed. Such tendencies may reflect the direction of gene flow between different races of maize: for instance, North American (Northern flint) and Caribbean populations were introduced, respectively, to northern and southern Europe, in the past. PMID- 12582567 TI - High-resolution mapping of the bolting gene B of sugar beet. AB - Sugar beet ( Beta vulgaris L.) is a biennial species. Shoot elongation (bolting) starts after a period of low temperature. The dominant allele of locus B causes early bolting without cold treatment. This allele is abundant in wild beets whereas cultivated beets carry the recessive allele. Fifteen AFLP markers, tightly linked to the bolting locus, have been identified using bulked segregant analysis. The F(2)-population consisted of 2,134 individuals derived after selfing a single F(1)-plant ( Bb). In a first step, a linkage map was established with 249 markers based on 775 F(2)-individuals with a coverage of 822.3 cM. The loci are dispersed over nine linkage groups corresponding to the haploid chromosome number of Beta species. Seventeen marker loci were placed at a distance less than 3.2 cM around the bolting gene. In a second step, four of those markers most closely linked to B were mapped with the entire F(2) population. Two of the markers were mapped flanking the B gene at distances of 0.14 and 0.23 cM. The other two markers were mapped at a distance of 0.5 cM from the gene. The tight linkage could be verified by testing 88 unrelated plants from a breeding program. The closely linked markers will enable breeders to select for the non-bolting character without laborious test crossings. Moreover, these markers are being used for map-based cloning of the bolting gene. PMID- 12582568 TI - Predicting within-family variability in juvenile height growth of Salix based upon similarity among parental AFLP fingerprints. AB - Willow is being developed as a crop for biomass plantations in the Northeast and North-central United States, but has only recently been the subject of controlled breeding to generate improved genotypes. Maximizing variability among progeny within full-sib families produced by controlled pollination may increase the probability of producing willow clones exhibiting desirable extreme phenotypes. Yet, predicting combinations of parents yielding highly variable progeny is not currently possible. Controlled pollinations were completed among 15 Salix eriocephala clones and the resulting progeny were vegetatively propagated and planted in a greenhouse progeny test. Heights of rooted cuttings were measured after 4 months of growth. Genetic similarity among parents was estimated based on 77 polymorphic AFLP bands. Strong negative correlation ( r = -0.88) was detected between mean female-parent similarity indices and the standard deviation of height among half-sib progeny from those females. Parent combinations that had relatively low similarity indices tended to produce progeny that had greater variability in height. This negative relationship suggests that AFLP fingerprints of S. eriocephala parents may be useful for predicting parent combinations that will yield families with large variability. PMID- 12582569 TI - Genomic regions controlling vernalization and photoperiod responses in oat. AB - Oat genotypes vary for photoperiod and vernalization responses. Vernalization often promotes earlier flowering in fall-sown but not spring-sown cultivars. Longer photoperiods also promote earlier flowering, and the response to longer photoperiods tends to be greater in cultivars from higher latitudes. To investigate the genetic basis of photoperiod and vernalization responses in oat, we mapped QTLs for flowering time under four combinations of photoperiod and vernalization treatments in the Ogle x TAM O-301 mapping population in growth chambers. We also mapped QTLs for flowering time in early spring and late-spring field plantings to determine the genetic basis of response to early spring planting in oat. Three major flowering-time QTLs (on linkage groups OT8, OT31 and OT32) were detected in most conditions. QTLs with smaller effects on flowering were less-consistently observed among treatments. Both vernalization-sensitive and insensitive QTLs were discovered. Longer photoperiod or vernalization alone tended to decrease the effects of flowering-time QTLs. Applied together, longer photoperiod and vernalization interacted synergistically, often on the same genomic regions. Earlier spring planting conferred an attenuated vernalization treatment on seeds. The major flowering-time QTLs mapped in this study matched those mapped previously in the Kanota x Ogle oat mapping population. Between these two studies, we found a concordance of flowering-time QTLs, segregation distortion, and complex genetic linkages. These effects may all be related to chromosomal rearrangements in hexaploid oat. Comparative mapping between oat and other grasses will facilitate molecular analysis of vernalization response in oat. PMID- 12582570 TI - Development of microsatellite markers in peach [ Prunus persica (L.) Batsch] and their use in genetic diversity analysis in peach and sweet cherry ( Prunus avium L.). AB - We report the sequence of 41 primer pairs of microsatellites from a CT-enriched genomic library of the peach cultivar 'Merrill O'Henry'. Ten microsatellite containing clones had sequences similar to plant coding sequences in databases and could be used as markers for known functions. For microsatellites segregating at least in one of the two Prunus F(2) progenies analyzed, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was evaluated in 27 peach and 21 sweet cherry cultivars. All primer pairs gave PCR-amplification products on peach and 33 on cherry (80.5%). Six PCR-amplifications revealed several loci (14.6%) in peach and eight (19.5%) in sweet cherry. Among the 33 single-locus microsatellites amplified in peach and sweet cherry, 13 revealed polymorphism both in peach and cherry, 19 were polymorphic only on peach and one was polymorphic only on cherry. The number of alleles per locus ranged from 1 to 9 for peach and from 1 to 6 on sweet cherry with an average of 4.2 and 2.8 in peach and sweet cherry, respectively. Cross-species amplification was tested within the Prunus species: Prunus avium L. (sweet cherry and mazzard), Prunus cerasus L. (sour cherry), Prunus domestica L. (European plum), Prunus amygdalus Batsch. (almond), Prunus armeniaca L. (apricot), Prunus cerasifera Ehrh. (Myrobalan plum). Plants from other genera of the Rosaceae were also tested: Malus (apple) and Fragaria (strawberry), as well as species not belonging to the Rosaceae: Castanea (chestnut tree), Juglans (walnut tree) and Vitis (grapevine). Six microsatellites gave amplification on all the tested species. Among them, one had an amplified region homologous to sequences encoding a MADS-box protein in Malus x domestica. Twelve microsatellites (29.3%) were amplified in all the Rosaceae species tested and 31 (75.6%) were amplified in all the six Prunus species tested. Thirty three (80.5%), 18 (43.9%) and 13 (31.7%) gave amplification on chestnut tree, grapevine and walnut tree, respectively. PMID- 12582571 TI - Combination of chloroplast and mitochondrial DNA polymorphisms to study cytoplasm genetic differentiation in the olive complex ( Olea europaea L.). AB - Four hundred and four individuals belonging to the species Olea europaea were characterised using mitochondrial DNA (mtDNA) RFLPs. Twelve mitotypes were distinguished. The combination of mtDNA information with cpDNA polymorphism (characterised in a previous study) led us to recognise 20 cytoplasmic lineages of which seven were found in the Mediterranean area (oleasters, cultivars and O. e. subsp. maroccana). In the olive complex, strong cytoplasm genetic differentiation was revealed ( F(st) = 0.73). Very strong linkage disequilibrium between cpDNA and mtDNA polymorphisms was observed, particularly in the Mediterranean subspecies europaea. This high congruence between genetic structure based on cpDNA or mtDNA sustains a low level of recurrent mutation in both organelle DNAs and, thus, the polymorphisms used in this study were pertinent to reconstruct olive phylogeography. In the Mediterranean area, genetic drift due to population regression during Quaternary glaciations, and founder effects associated with the postglacial seed dissemination, have probably contributed to the existence of a high genetic linkage disequilibrium between cpDNA and mtDNA polymorphisms. Thus, four Mediterranean cytoplasmic lineages, clearly distinguished both by cpDNA and mtDNA polymorphisms, most likely reflect four distinct relic populations during Quaternary glaciations. Finally, O. e. subsp. maroccana from South Morocco, which also displayed specific cytoplasmic lineages, should be considered as another relic Mediterranean population. PMID- 12582572 TI - Candidate genes and QTLs for sugar and organic acid content in peach [ Prunus persica (L.) Batsch]. AB - The identification of genes involved in variation of peach fruit quality would assist breeders in creating new cultivars with improved fruit quality. Major genes and quantitative trait loci (QTLs) for physical and chemical components of fruit quality have already been detected, based on the peach [ Prunus persica (L.) Batsch] cv. Ferjalou Jalousia((R)) (low-acid peach) x cv. Fantasia (normally acid nectarine) F(2) intraspecific cross. Our aim was to associate these QTLs to structural genes using a candidate gene/QTL approach. Eighteen cDNAs encoding key proteins in soluble sugar and organic acid metabolic pathways as well as in cell expansion were isolated from peach fruit. A single-strand conformation polymorphism strategy based on specific cDNA-based primers was used to map the corresponding genes. Since no polymorphism could be detected in the Ferjalou Jalousia((R)) x Fantasia population, gene mapping was performed on the almond [ Prunus amygdalus ( P. dulcis)] cv. Texas x peach cv. Earlygold F(2) interspecific cross from which a saturated map was available. Twelve candidate genes were assigned to four linkage groups of the peach genome. In a second step, the previous QTL detection was enhanced by integrating anchor loci between the Ferjalou Jalousia((R)) x Fantasia and Texas x Earlygold maps and data from a third year of trait assessment on the Ferjalou Jalousia((R)) x Fantasia population. Comparative mapping allowed us to detect a candidate gene/QTL co location. It involved a cDNA encoding a vacuolar H(+)-pyrophosphatase ( PRUpe;Vp2) that energises solute accumulation, and QTLs for sucrose and soluble solid content. This preliminary result may be the first step in the future development of marker-assisted selection for peach fruit sucrose and soluble solid content. PMID- 12582573 TI - Development of a mungbean ( Vigna radiata) RFLP linkage map and its comparison with lablab ( Lablab purpureus) reveals a high level of colinearity between the two genomes. AB - A genetic linkage map of mungbean ( Vigna radiata, 2n = 2 x = 22) consisting of 255 RFLP loci was developed using a recombinant inbred population of 80 individuals. The population was derived from an inter-subspecific cross between the cultivated mungbean variety 'Berken' and a wild mungbean genotype 'ACC 41' ( V. radiata subsp. sublobata). The total length of the map, which comprised 13 linkage groups, spanned 737.9 cM with an average distance between markers of 3.0 cM and a maximum distance between linked markers of 15.4 cM. The mungbean map was compared to a previously published map of lablab ( Lablab purpureus, 2n = 2 x = 24) using a common set of 65 RFLP probes. In contrast to some other comparative mapping studies among members of the Fabaceae, where a high level of chromosomal rearrangement has been observed, marker order between mungbean and lablab was found to be highly conserved. However, the two genomes have apparently accumulated a large number of duplications/deletions after they diverged. PMID- 12582574 TI - QTL clusters reflect character associations in wild and cultivated rice. AB - The genetic basis of character association related to differentiation found in the primary gene pool of rice was investigated based on the genomic distribution of quantitative trait loci (QTLs). Major evolutionary trends in cultivated rice of Asiatic origin ( Oryza sativa) and its wild progenitor ( O. rufipogon) are: (1) differentiation from wild to domesticated types (domestication), (2) ecotype differentiation between the perennial and annual types in wild races, and (3) the Indica versus Japonica type differentiation in cultivated races. Using 125 recombinant inbred lines (RILs) derived from a cross between an Indica cultivar of O. sativa and a strain of O. rufipogon carrying some Japonica-like characteristics, we mapped 147 markers, mostly RFLPs, on 12 chromosomes. Thirty seven morphological and physiological quantitative traits were evaluated, and QTLs for 24 traits were detected. The mapped loci showed a tendency to form clusters that are composed of QTLs of the domestication-related traits as well as Indica/Japonica diagnostic traits. QTLs for perennial/annual type differences did not cluster. This cluster phenomenon could be considered "multifactorial linkages" followed by natural selection favoring co-adapted traits. Further, it is possible that the clustering phenomenon is partly due to pleiotropy of some unknown key factor(s) controlling various traits through diverse metabolic pathways. Chromosomal regions where QTL clusters were found coincided with the regions harboring genes or gene blocks where the frequency of cultivar-derived alleles in RILs is higher than expected. This distortion may be partly due to unconscious selection favoring cultivated plant type during the establishment of RILs. PMID- 12582575 TI - Diversity of Olea genotypes and the origin of cultivated olives. AB - Tandem repeats belonging to three DNA sequence families ( OeTaq80, OeTaq178, and OeGEM86) were isolated from the nuclear DNA of Olea europaea cv. Carolea and dot hybridized to the genomic DNA of 14 hypothetically different Olea species, 78 olive cultivars, and 14 wild olives. The copy number per unreplicated haploid genome of OeTaq80- and OeTaq178-related sequences was in the 10(7)-10(6) range and that of OeGEM86-related sequences was in the 10(5) range in cultivars, wild olives and some Olea species. A large variation in the frequency of repeats belonging to each sequence family was observed within each group of plants. Positive correlations existed in each genome between the frequencies of repeats belonging to each family, and their overall frequency was positively correlated to the genome size. Duncan grouping showed that the frequency variation of tandem repeats within each group of plants was not continuous. Two main groups and several subgroups of genotypes could be separated within both the olive cultivars and the wild olives. Discrete areas in the Mediterranean Basin could be delimited by the geographic distribution of cultivated olives with different genotypes and the wild plants were associated with the cultivars in these areas according to genotypic similarity. The Olea species could be divided into four genotypic groups. Three of these, comprising accessions from Asia and North Africa, showed similarity with the genotypes of cultivars and wild olives. These results suggest a polyphyletic origin of cultivated olives from different wild Olea forms distributed throughout the Mediterranean Basin. PMID- 12582576 TI - Analysis of genetic diversity and sectional relationships in Musa using AFLP markers. AB - The AFLP technique was used to assess the genetic diversity and sectional relationships in 39 accessions representing the four main sections of the genus Musa. Eight AFLP + 3 primer pairs produced 260 polymorphic bands that were used in cluster and PCO analysis. A wide range of variability was observed among the species within the sections of the genus Musa. AFLP data was useful in separating the different sections of the genus as well as differentiating the different genomic groups of section Eumusa. Section Rhodochlamys ( x = 11) appeared as a distinct entity and clustered closely with the Musa acuminata Colla complex of section Eumusa that has the same basic chromosome number. This relationship is congruent with previous studies. However, unlike previous proposals that questioned the identity of Rhodochlamys as a separate taxonomic unit, PCO analysis of the AFLP data showed that it is a distinct entity. Musa laterita Cheesman ( Rhodochlamys) and Musa schizocarpa Simmonds clustered with the M. acuminata complex suggesting that they may be sources of useful genes for the improvement of the cultivated bananas. Callimusa formed a distinct unit and was closer to Australimusa than to the other sections. Although both sections share the same basic chromosome number of x = 10 these sections are genetically distinct PMID- 12582577 TI - Genetic Diversity in Musa acuminata Colla and Musa balbisiana Colla and some of their natural hybrids using AFLP Markers. AB - Genetic diversity and relationships were assessed in 28 accessions of Musa acuminata (AA) Colla and Musa balbisiana (BB) Colla, and some of their natural hybrids, using the amplified fragment length polymorphisms (AFLP) technique. Fifteen AFLP +3 primer pairs produced 527 polymorphic bands among the accessions. Neighbor-joining and principal co-ordinate (PCO) analyses using Jaccard's similarity coefficient produced four major clusters that closely corresponded with the genome composition of the accessions (AA, BB, AAB and ABB). The AFLP data distinguished between the wild diploid accessions and suggested new subspecies relationships in the M. acuminata complex that are different from those based on morphological data. The data suggested that there are three subspecies within the M. acuminata complex (ssp. burmannica Simmonds, malaccensis Simmonds, and microcarpa Simmonds). 'Tjau Lagada' (ssp. microcarpa), 'Truncata' [ssp truncata (Ridl.) Shepherd] and 'SF247' [ssp. banksii (F.Muell) Simmonds] clustered very closely with 'Gros Michel' and 'Km 5', indicating that more than one M. acuminata subspecies may be involved in the origin of triploid AAA bananas. 'Calcutta 4' (ssp. burmannicoides De Langhe & Devreux) and 'Long Tavoy' (ssp. burmannica) were closely related and could be together in the same subspecies. This study also showed that there is much more genetic diversity within M. balbisiana that was split into two groups: (1) 'I-63' and 'HND' and (2) 'Los Banos', 'MPL' (Montpellier), '10852', 'Singapuri', 'Etikehel', and 'Butohan 1' as the other. PMID- 12582578 TI - Identification and classification of S haplotypes in Raphanus sativus by PCR-RFLP of the S locus glycoprotein (SLG) gene and the S locus receptor kinase (SRK) gene. AB - Polymorphism of the S-locus glycoprotein (SLG) and S-locus receptor kinase (SRK) genes in Raphanus sativus was analyzed by PCR-RFLP using SLG- and SRK-specific primers. Twenty four inbred lines of R. sativus could be grouped into nine S haplotypes. DNA fragments of SLG alleles specifically amplified from five S haplotypes by PCR with Class-I SLG-specific primers showed different profiles upon polyacrylamide-gel electrophoresis after digestion with restriction endonucleases. The five R. sativus SLG alleles were determined for their nucleotide sequences of DNA fragments. Comparison of the amino-acid sequences with a reported Brassica SLG (S(6)) showed 77-84% homology. Deduced amino-acid sequences showed 12-conserved cystein residues and three hypervariable regions which are characteristic of Brassicsa SLG. A DNA fragment was also amplified by PCR from two of each S haplotype with Class-II SLG-specific primers, and showed polymorphism when cleaved with restriction endonucleases. The nucleotide sequences of amplified DNA fragments of the Class-II SLG revealed about 60% similarity with those of the Class-I SLG. It is concluded that there exist both Class I and Class II S alleles in R. sativus, as in Brassica campestris and Brassica oleracea. PCR using SRK-specific primers amplified a DNA fragment of about 1.0 kb from seven of each S haplotype out of 24 tested. These DNA fragments showed high polymorphism in polyacrylamide-gel electrophoresis after digestion with restriction endonucleases. Nucleotide sequences of the DNA fragments amplified from the seven S haplotypes showed that the fourth and the fifth exons of SRK are highly conserved, and that there is high variation in the fifth intron, the sixth intron and seventh exon of the SRK which may be responsible for the polymorphic band patterns in PCR-RFLP analysis. The PCR-RFLP method has proven useful for the identification of S alleles in inbred lines and for listing S haplotypes in R. sativus. Phylogenic analysis of the SLG and SRK sequences from Raphanus and Brassica revealed that the Raphanus SLGs and SRKs did not form an independent cluster, but were dispersed in the tree, clustering together with Brassica SLGs and SRKs. Furthermore, SLGs and SRKs from Raphanus were both grouped into Class-I or Class-II S haplotypes. Therefore, these results suggest that the diversification of the SLG and SRK alleles occurred prior to the differentiation of the two genera Brassica and Raphanus. PMID- 12582579 TI - Methylation-sensitive RFLPs: characterisation of two oil palm markers showing somaclonal variation-associated polymorphism. AB - The occurrence of "mantled" somaclonal variants (approx. 5%), which display alterations in floral organ structure, among populations of somatic embryo derived oil palms ( Elaeis guineensis Jacq.) currently hampers any scaling-up of clonal plant micropropagation. As a first step towards the identification of abnormality-discriminating markers, we have screened a set of 27 oil palm cDNA probes for methylation-sensitive restriction fragment length polymorphisms (RFLPs) using callus genomic DNA digested with the isoschizomeric enzymes MspI and HpaII. Only two probes (CPHO62 and -63) were found to differentiate reproducibly in two different genotypic backgrounds between nodular compact calli (NCC) and fast-growing calli (FGC), which generate 5% and 100% "mantled" plantlets, respectively. Comparative analyses were then conducted on DNA from inflorescences and leaves of normal and abnormal adult regenerants. With both probes, the observed methylation patterns were strongly clone-dependent and monomorphic with respect to the phenotype of the regenerants, except for the type specific banding pattern obtained with the CPHO62 probe on material from the LMC3 clonal offspring. The results presented here mirror the higher difference in genomic DNA methylation observed between normal and abnormal embryogenic calli when compared to more differentiated plant material. Moreover, they reinforce the paramount interest of NCC and FGC callus lines as a material of choice in the search for early epigenetic markers of the "mantled" somaclonal variation. The potential use of methylation-sensitive RFLPs for the early detection of somaclonal variation at early stages of the micropropagation process is discussed. PMID- 12582580 TI - SSR analysis of genetic diversity among maize inbred lines adapted to cold regions of Japan. AB - Information regarding diversity and relationships among breeding material is necessary for hybrid maize ( Zea mays L.) breeding. Simple-sequence repeat (SSR) analysis of the 60 loci distributed uniformly throughout the maize genome was carried out for 65 inbred lines adapted to cold regions of Japan in order to assess genetic diversity among the inbred lines and to assign them to heterotic groups. The mean value (0.69) of the polymorphic-index content (PIC) for the SSR loci provided sufficient discrimination-ability for the assessment of genetic diversity among the inbred lines. The correlation between the genetic-similarity (GS) estimates and the coancestry coefficient was significant ( r = 0.70). The average-linkage (UPGMA) cluster analysis and principal-coordinate analysis (PCOA) for a matrix of the GS estimates showed that the Northern flint inbred lines bred in Japan were similar to a Canadian Northern flint inbred line CO12 and a European flint inbred line F283, and that dent inbred lines bred in Japan were similar to BSSS inbred lines such as B73. These associations correspond to the known pedigree records of these inbred lines. The results indicate that SSR analysis is effective for the assessment of genetic diversity among maize inbred lines and for the assignment of inbred lines to heterotic groups. PMID- 12582581 TI - The development of a STS marker linked to a yellow rust resistance derived from the wheat cultivar Moro. AB - A sequence-tagged-site (STS) marker has been developed for a gene conferring yellow rust resistance originating from the wheat cultivar Moro. The single, dominant, seedling yellow rust resistance gene, designated YrMoro, was mapped to the group 1 chromosomes. The STS marker was developed from an AFLP band which cosegregated with the YrMoro gene. Sequence-specific primers were made which incorporated the selective bases of the AFLP primers, plus 16 and 17 additional bases extending into the AFLP band. This simple, PCR-based marker will allow wheat breeders to pyramid this resistance gene, along with other resistance genes, into a single wheat genotype. PMID- 12582582 TI - Isolation from alfalfa of resistance gene analogues containing nucleotide binding sites. AB - Cloned resistance (R) genes from a broad range of plant species are known to share similarities in DNA sequence and structural motifs. Degenerate oligonucleotide primers designed from conserved regions of the nucleotide binding site (NBS), common to many R genes, were used to amplify the NBS regions from genomic DNA from alfalfa ( Medicago sativa L). Sequence comparisons of the amplified fragments indicated that at least 18 families of NBS-containing R genes are present in alfalfa. Comparisons to R genes from other species suggested a polyphyletic origin of these gene families. Using the same degenerate primers, PCR analysis of cDNA prepared from a plant not challenged with a pest or pathogen revealed that many of the NBS-containing gene families were transcribed actively. Amplification of NBS regions from other Medicago species showed the presence of some NBS-containing genes not present in alfalfa. These results indicate that the NBS-containing R genes comprise a large gene family in Medicago, at least some of which are transcribed in healthy plants, and that different Medicago species carry unique NBS genes. PMID- 12582583 TI - Seed gene flow and fine-scale structure in a Mediterranean pine ( Pinus pinaster Ait.) using nuclear microsatellite markers. AB - The Mediterranean populations of maritime pine ( Pinus pinaster Ait.) are typically small and have a scattered distribution, being threatened by human activities and forest fires. In the framework of the genetic-resources conservation program of this species, a native multi-age stand located in a Mediterranean area (central Spain) was studied using three highly polymorphic nuclear microsatellites (SSRs). Spatial autocorrelation analysis was conducted using Moran's index in order to detect fine-scale structure in both natural regeneration and mature trees. The spatial pattern of seed flow based on dispersed progeny was studied using a highly reliable subset of parent-offspring matches obtained by means of parentage analysis and simulation-based calculation of statistical confidence. Maritime pine showed a fine-scale structure at the seedling stage. In natural regeneration, the autocorrelograms indicated a patch size of approximately 10 m. The fine-scale structure seems to be produced by a restricted seed gene flow. In fact, there was an excess of parent-offspring matches in a radius of 15 m from the parent trees. Pines with a heavy seed, such as P. pinaster, are expected to have a short dispersal distance, thus producing a fine-scale structure. However, the fine-scale structure did not persist in the mature trees. Within-population genetic structure in Mediterranean pines may be affected by a number of post-dispersal events (e.g. mortality due to the severity of the Mediterranean climate and animal-mediated secondary dispersal during the summer period). Thus, great alteration in the pattern produced by the initial seed rain and differences in genetic structure between tree cohorts are expected. PMID- 12582584 TI - A barley gene family homologous to the maize rust resistance gene Rp1-D. AB - Many characterized plant disease resistance genes encode proteins which have conserved motifs such as the nucleotide binding site. Conservation extends across different species, therefore resistance genes from one species can be used to isolate homologous regions from another by employing DNA sequences encoding conserved protein motifs as probes. Here we report the isolation and characterization of a barley ( Hordeum vulgare L.) resistance gene analog family consisting of nine members homologous to the maize rust resistance gene Rp1-D. Five barley Rp1-D homologues are clustered within approximately 400 kb on chromosome 1(7H), near, but not co-segregating with, the barley stem rust resistance gene Rpg1; while others are localized on chromosomes 3(3H), 5(1H), 6(6H) and 7(5H). Analyses of predicted amino-acid sequences of the barley Rp1-D homologues and comparison with known plant disease resistance genes are presented. PMID- 12582585 TI - Paternal chloroplast inheritance patterns in pine hybrids detected with trnL-trnF intergenic region polymorphism. AB - The inheritance patterns of the chloroplast genomes of shortleaf pine ( Pinus echinata Mill.), loblolly pine ( Pinus taeda L.) and slash pine ( Pinus elliottii Engelm.) were investigated through the trnL-trnF intergenic spacer polymorphism analysis. The DNA sequences of this spacer differ among these three closely related Pinus species. A modified 'cold' PCR-SSCP (single-strand conformation polymorphism) analysis of this spacer shows that the artificial hybrids (F1) from the shortleaf pine (seed parent) x loblolly pine (pollen parent) cross, exhibit the loblolly pine profile. Additionally, nine putative hybrids between shortleaf pine and loblolly pine, previously identified by the IDH (Isocitrate dehydrogenase) allozyme marker, presented the shortleaf pine profile indicating that shortleaf pine, not loblolly pine, sired all of the putative hybrids. Nondenatured polyacrylamide-gel electrophoresis of the trnL-trnF intergenic spacer demonstrated that the artificial hybrids (F1) from the cross, slash pine (seed parent) x shortleaf pine (pollen parent), present the shortleaf pine profile. Those results confirmed that the chloroplast genome is paternally inherited in these three species of the genus Pinus. The significance of the trnL trnF intergenic region polymorphism and our modified 'cold' SSCP protocol for population genetic studies is discussed. PMID- 12582586 TI - Mapping of the nodulation loci sym9 and sym10 of pea ( Pisum sativum L.). AB - Several mutants defective in the nodulation process during rhizobial or endomycorrhizal endosymbiosis of pea have been identified previously. We have integrated the map positions of two such nodulation mutations, sym9 and sym10, into the molecular map of pea by applying molecular-marker techniques combined with bulked segregant analysis (BSA). Lines P2 and P54 were found to carry alleles of sym9, line P56 carried an allele of sym10. F2 populations were derived from crosses of P2, P54 and P56, to JI281 and JI15, two of the parental lines that have been used previously to generate a molecular map of pea. sym9 was located on linkage group IV by AFLP-BSA analysis and subsequently mapped by RFLP in both F2 populations, P2 x JI281 and P54 x JI281. RFLP-BSA analysis was applied to assign sym10 to linkage group I. The RFLP marker locus, chs2, co-segregates with sym10 in the F2 population of P56 x JI15. PMID- 12582587 TI - Identification and mapping of molecular markers linked to rust resistance genes located on chromosome 1RS of rye using wheat-rye translocation lines. AB - The short arm of rye ( Secale cereale) chromosome 1 has been widely used in breeding programs to incorporate new disease resistance genes into wheat. Using wheat-rye translocation and recombinant lines, molecular markers were isolated and mapped within chromosomal regions of 1RS carrying rust resistance genes Lr26, Sr31, Yr9 from 'Petkus' and SrR from 'Imperial' rye. RFLP markers previously mapped to chromosome 1HS of barley - flanking the complex Mla powdery mildew resistance gene locus - and chromosome 1DS of Aegilops tauschii - flanking the Sr33 stem rust resistance gene - were shown to map on either side of rust resistance genes on 1RS. Three non cross-hybridising Resistance Gene Analog markers, one of them being derived from the Mla gene family, were mapped within same region of 1RS. PCR-based markers were developed which were tightly linked to the rust resistance genes in 'Imperial' and 'Petkus' rye and which have potential for use in marker-assisted breeding. PMID- 12582588 TI - Optimal sampling strategy and core collection size of Andean tetraploid potato based on isozyme data - a simulation study. AB - Selection of an appropriate sampling strategy is an important prerequisite to establish core collections of appropriate size in order to adequately represent the genetic spectrum and maximally capture the genetic diversity in available crop collections. We developed a simulation approach to identify an optimal sampling strategy and core-collection size, using isozyme data from a CIP germplasm collection on an Andean tetraploid potato. Five sampling strategies, constant (C), proportional (P), logarithmic (L), square-root (S) and random (R), were tested on isozyme data from 9,396 Andean tetraploid potato accessions characterized for nine isozyme loci having a total of 38 alleles. The 9,396 accessions, though comprising 2,379 morphologically distinct accessions, were found to represent 1,910 genetically distinct groups of accessions for the nine isozyme loci using a sort-and-duplicate-search algorithm. From each group, one accession was randomly selected to form a genetically refined entire collection (GREC) of size 1,910. The GREC was used to test the five sampling strategies. To assess the behavior of the results in repeated sampling, k = 1,500 and 5,000 independent random samples (without replacement) of admissible sizes n = 50(50)1,000 for each strategy were drawn from GREC. Allele frequencies (AF) for the 38 alleles and locus heterozygosity (LH) for the nine loci were estimated for each sample. The goodness of fit of samples AF and LH with those from GREC was tested using the chi(2) test. A core collection of size n = 600, selected using either the P or the R sampling strategy, was found adequately to represent the GREC for both AF and LH. As similar results were obtained at k = 1,500 and 5,000, it seems adequate to draw 1,500 independent random samples of different sizes to test the behavior of different sampling strategies in order to identify an appropriate sampling approach, as well as to determine an optimal core collection size. PMID- 12582589 TI - Genetic differentiation for nuclear, mitochondrial and chloroplast genomes in common wild rice ( Oryza rufipogon Griff.) and cultivated rice ( Oryza sativa L.). AB - The genetic differentiation of nuclear, mitochondrial (mt) and chloroplast (cp) genomes was investigated by Southern and PCR analysis using 75 varieties of cultivated rice ( Oryza sativa L.) and 118 strains of common wild rice (CWR, Oryza rufipogon Griff.) from ten countries of Asia. The distinguishing differences between the Indica and Japonica cultivars were detected both in the nuclear genome and the cytoplasmic genome, confirming that the Indica-Japonica differentiation is of major importance for the three different classes of genome in cultivated rice. This differentiation was also detected in common wild rice with some differences among the genome compartments and the various regions. For nuclear DNA variation, both Indica-like and Japonica-like types were observed in the Chinese CWR, with the latter more-frequent than the former. No Japonica-like type was found in South Asia, and only two strains of the Japonica-like type were detected in Southeast Asia, thus the Indica-like type is the major type among South and Southeast Asian CWR. For mtDNA, only a few strains of the Japonica-like type were detected in CWR. For cpDNA, the Japonica type was predominant among the CWR strains from China, Bangladesh and Burma, while the Indica type was predominant among the CWR strains from Thailand, Malaysia, Cambodia and Sri Lanka, and both types were found in similar frequencies among the Indian CWR. Altogether, however, the degree of Indica-Japonica differentiation in common wild rice was much-less important than that in cultivated rice. Cluster analyses for nuclear and mitochondrial DNA variation revealed that some CWR strains showed large genetic distances from cultivated rice and formed clusters distinct from cultivated rice. Coincidence in the genetic differentiation between the three different classes of genome was much higher in cultivated rice than in CWR. Among the 75 cultivars, about 3/4 entries were "homoeotype" showing congruent results for nuclear, mt and cpDNA regarding the Indica-Japonica differentiation. In CWR, the proportions of homoeotypes were 5.7%, 15% and 48.8% in China, South Asia and Southeast Asia, respectively. Based on the average genetic distance among all the strains of CWR and cultivated rice for nuclear and mitochondrial genomes, the variability of the nuclear genome was found to be higher than that of the mitochondrial genome. The global pattern based on all genomes shows much-more diversification in CWR than that in cultivated rice. PMID- 12582590 TI - Structure and level of genetic diversity in various bean types evidenced with microsatellite markers isolated from a genomic enriched library. AB - We have constructed a common bean genomic library enriched for microsatellite motifs (ATA), (CA), (CAC) and (GA). After screening, 60% of the clones selected from the library enriched for the (ATA) repeat contained microsatellites versus 21% of the clones from the library enriched for (GA) (CA) and (CAC) repeats. Fifteen primer pairs have been developed allowing for the amplification of SSR loci. We have evaluated the genetic diversity of these loci between 45 different bean lines belonging to nine various quality types. A total of 81 alleles were detected at the 15 microsatellite loci with an average of 5.3 alleles per locus. We have investigated the origin of allelic size polymorphism at the locus PvATA20 in which the number of repeats ranges from 24 to 85. We have related these large differences in repeat number to unequal crossing-over between repeated DNA regions. The diversity analysis revealed contrasted levels of variability according to the bean type. The lower level was evidenced for the very fine French bean, showing the effect of breeders intensive selection. PMID- 12582591 TI - Olea europaea (Oleaceae) phylogeography based on chloroplast DNA polymorphism. AB - Chloroplast DNA diversity in the olive ( Olea europaea L.) complex was studied using PCR-RFLP and microsatellite markers. Fifteen chlorotypes were distinguished. We constructed a cpDNA phylogenetic tree in which five clades were recognised and located in distinct geographic areas: clade A in Central and Southern Africa, clade C in Asia, clade M in North-West Africa, clade E1 in the Mediterranean Basin and Sahara, and clade E2 in West Mediterranea. Cultivated olive clustered with Mediterranean and Saharan wild forms (clades E1 and E2). Strong genetic differentiation for cpDNA markers was observed between eastern and western Mediterranean olives, suggesting that these areas have represented different glacial refugia. Humans most likely spread one eastern chlorotype, preponderant in cultivars, across the western Mediterranean Basin. Its presence in O. e. subsp. laperrinei from the Sahara suggests a possible Mediterranean olive origin in an African population, which may have overlapped in the Southern Mediterranean during the Quaternary. PMID- 12582592 TI - Flow karyotyping and chromosome sorting in bread wheat ( Triticum aestivum L.). AB - Previously, we reported on the development of procedures for chromosome analysis and sorting using flow cytometry (flow cytogenetics) in bread wheat. That study indicated the possibility of sorting large quantities of intact chromosomes, and their suitability for analysis at the molecular level. However, due to the lack of sufficient differences in size between individual chromosomes, only chromosome 3B could be sorted into a high-purity fraction. The present study aimed to identify wheat stocks that could be used to sort other chromosomes. An analysis of 58 varieties and landraces demonstrated a remarkable reproducibility and sensitivity of flow cytometry for the detection of numerical and structural chromosome changes. Changes in flow karyotype, diagnostic for the presence of the 1BL.1RS translocation, have been found and lines from which translocation chromosomes 5BL.7BL and 4AL.4AS-5BL could be sorted have been identified. Furthermore, wheat lines have been identified which can be used for sorting chromosomes 4B, 4D, 5D and 6D. The ability to sort any single arm of the hexaploid wheat karyotype, either in the form of a ditelosome or a isochromosome, has also been demonstrated. Thus, although originally considered recalcitrant, wheat seems to be suitable for the development of flow cytogenetics and the technology can be applied to the physical mapping of DNA sequences, the targeted isolation of molecular makers and the construction of chromosome- and arm specific DNA libraries. These approaches should facilitate the analysis of the complex genome of hexaploid bread wheat. PMID- 12582593 TI - Genetic mapping of QTLs associated with greenbug resistance and tolerance in Sorghum bicolor. AB - Ninety three recombinant inbreds of Sorghum bicolor (L. Moench) were derived from a cross between two sorghum lines GBIK and Redlan. This population was used to identify quantitative trait loci (QTLs) for resistance and tolerance to greenbug ( Schizaphids graminum Rondani) Biotypes I and K. One hundred and thirteen loci (38 SSRs and 75 RAPDs) were mapped in 12 linkage groups covering 1,530 cM. In general, nine QTLs were detected affecting both resistance and tolerance to greenbug (GB) Biotypes I and K. The phenotypic variance explained by each QTL ranged from 5.6% to 38.4%. Four SSRs and one RAPD marker were associated with the expression of all resistance and tolerance traits. These markers appear to be linked to biotype non-specific resistance and tolerance genes. Four additional markers were associated with biotype-specific resistance or tolerance traits. The detection of more than one locus for each biotype supports the hypothesis that several regions, which represent different genes, control the expression of resistance and tolerance to greenbug in sorghum. The results can be used for marker-assisted selection and the breeding of greenbug-tolerant sorghum cultivars. PMID- 12582594 TI - Genome plasticity in Festuca arundinacea: direct response to temperature changes by redundancy modulation of interspersed DNA repeats. AB - The response of the genome of Festuca arundinacea seedlings to changes in the temperature at which they were grown was investigated. Fifteen repeated sequences in the nuclear DNA were isolated and hybridized to the genomic DNA of seedlings grown at 10 degrees C or 30 degrees C. The redundancies of sequences recognized by four probes ( FaA5, FaH8, FaH13 and FaH14), were found to differ significantly in the two DNAs. DNA sequences recognized by FaH8, FaH13 and FaH14 were more represented in the genome of the 30 degrees C-raised seedlings than in the genome of the 10 degrees C-raised seedlings (76.5 x 10(3), 1.9 x 10(3), and 111.8 x 10(3) copies per haploid, 1C genome vs 62.7 x 10(3), 1.3 x 10(3), and 80.8 x 10(3) copies, respectively). In contrast, FaA5-related sequences were more represented in the genome of seedlings grown at the lower temperature (15.5 x 10(3) vs 10.2 x 10(3) copies, respectively). Southern-blot hybridization of these repeats to digested genomic DNA produced patterns which indicated that the probe sequences were part of longer repeated sequences having a limited degree of structural heterogeneity. These patterns were partly different when the probes were hybridized to the DNA from seedlings grown at 10 degrees C or 30 degrees C. In situ hybridization showed that the DNA sequences recognized by each probe were scattered along the length of all the chromosomes, with preferential location of FaA5- and FaH13-related sequences at given, mainly centromeric, regions of certain chromosomes. These findings suggest that redundancy modulations of interspersed repeated sequences allow direct responses of the genome of F. arundinacea to changes in environmental temperature. PMID- 12582595 TI - Comparison between genetic and physical maps in Zea mays L. of molecular markers linked to resistance against Diatraea spp. AB - In the pachytene stage, chromosomes are maximally extended and can easily be distinguished. Therefore, by applying fluorescence in situ hybridization (FISH) to pachytene chromosomes, it is possible to generate a high-resolution physical map of chromosome 9 in maize. Molecular markers ( umc105a on the short arm of chromosome 9, csu145a on the long arm) were used that flank quantitative trait loci (QTL) for sugarcane borer (SCB) and southwestern corn borer (SWCB) resistance. As reference markers, a centromere-specific probe (CentC) and a knob specific probe (pZm4-21) were utilized. Two fluorescent dyes with four probes were used to physically position these markers. Signals of repetitive DNA sequences in cosmid probes were suppressed by chromosome in situ suppression (CISS) hybridization. FISH signals were strong and reproducible for all probes. We measured the distances in micrometers for four subchromosomal regions and estimated the corresponding number of base pairs. The physical locations of the markers were compared on mitotic metaphase and pachytene chromosomes to the genetic map of chromosome 9. Genetic analysis positioned the two markers for SCB resistance in a central interval representing approximately 33.7% of the genetic length. However, the physical distance between these probes was determined to encompass about 70% of the physical length of chromosome 9. The two markers were located at distal positions on opposite arms of chromosome 9. Physical maps provide valuable information for gene isolation and understanding recombination. PMID- 12582596 TI - Insertional tagging of regulatory sequences in tritordeum; a hexaploid cereal species. AB - As an approach to isolate novel cereal promoters, promoterless uidA constructs and particle bombardment were used to transform tritordeum. Five of eight transgenic lines containing uidA sequences showed evidence of promoter tagging. Expression of uidA was detected in four lines as: constitutive expression, expression in short cells of the epidermis of the spikelets, expression in pollen grains and in cells of the epidermis of the spikelet, and expression in anther primordia and pollen grains. In the fifth line, the uidA was shown by RT-PCR to be transcribed, but no GUS activity was detected. The different patterns of uidA expression indicate that different regulatory sequences were tagged in each of these lines. Analysis of the progeny resulting from self-fertilisation of the primary tagged plants, indicate that the transgenes integrated at one or two loci and the patterns of expression were stably inherited. To our knowledge, this is the first report of promoter tagging in cereals by direct gene transfer. PMID- 12582597 TI - Genomic and chromosomal organization of Ty1- copia-like sequences in Olea europaea and evolutionary relationships of Olea retroelements. AB - The Ty1- copia-like retrotransposon is one of the commonest class of transposable elements in the plant kingdom, often comprising several percent of the total DNA content. We aimed to study the evolutionary relationships of Olea retroelements, using part of the reverse transcriptase domain, as well as the genomic and chromosomal organization of these sequences in Olea europaea chromosomes and their transcription activity and copy number. Fourteen clones, that were isolated from four different species, were sequenced and a phylogenetic tree was constructed based on their predicted amino acids. Five clones derived from O. europaea were clustered together with a 87% nucleotide sequence homology and two Olea oleaster clones showed 98% sequence homology. The rest of the clones showed heterogeneity among them, leading to a common ancestral transposon that existed before the genus arose. The Ty1- copia-like sequences have a dispersed genomic organization, physically distributed on all chromosomes, showing minor clustering in some cases and low copy numbers in the smallest chromosome pair. The total copy number in the O. europaea genome was estimated by dot blotting to be 40,000 in a haploid nucleus, but a number of these are non-functional since the sequenced clones contained stop codons and frame-shifts. Some Ty1- copia-like copies, present in O. europaea, were found to be methylated, while no differences in methylation were observed between DNA isolated from young leaves and callus suspension cultures. PMID- 12582598 TI - Allozyme evidence supporting southwestern Europe as a secondary center of genetic diversity for the common bean. AB - Genetic diversity within a common bean ( Phaseolus vulgaris L.) collection, comprising 343 accessions from the Iberian Peninsula, was examined using six allozyme markers. Two major clusters corresponding to the Andean and Mesoamerican gene pools were identified. Both gene pools were characterized by specific alleles, with the former exhibiting Skdh(100), Me(100), Rbcs(100 or 98) and Diap 1(100), and the latter exhibiting Skdh(103), Me(100), Rbcs(100) and Diap-1(95). Some accessions from both clusters, deviating from these allozyme patterns, exhibited Skdh(100), Me(100), Rbcs(100) and Diap-1(95) or Skdh(103), Me(100), Rbcs(100) and Diap-1(100) allozyme profiles and were considered as putative hybrids.The levels of genetic variation has not been eroded since the introduction of the common bean from the American centers of domestication to the Iberian Peninsula. Instead, obvious signs of introgression between the two gene pools were observed, mainly among white-seeded genotypes. The intermediate forms adapted to the Iberian Peninsula could have emerged from initial recombination between Mesoamerican and Andean gene pools. The Iberian common bean germplasm is therefore more complex than previously thought, and contains additional diversity that remains to be explored for genetic and breeding purposes. The Iberian Peninsula could be considered as a secondary center of genetic diversity of the common bean, especially the large white-seeded genotypes. PMID- 12582599 TI - Identification of QTLs for early blight ( Alternaria solani) resistance in tomato using backcross populations of a Lycopersicon esculentum x L. hirsutum cross. AB - Most commercial cultivars of tomato, Lycopersicon esculentum Mill., are susceptible to early blight (EB), a devastating fungal ( Alternaria solani Sorauer) disease of tomato in the northern and eastern parts of the U.S. and elsewhere in the world. The disease causes plant defoliation, which reduces yield and fruit quality, and contributes to significant crop loss. Sources of resistance have been identified within related wild species of tomato. The purpose of this study was to identify and validate quantitative trait loci (QTLs) for EB resistance in backcross populations of a cross between a susceptible tomato breeding line (NC84173; maternal and recurrent parent) and a resistant Lycopersicon hirsutum Humb. and Bonpl. accession (PI126445). Sixteen hundred BC(1) plants were grown to maturity in a field in 1998. Plants that were self incompatible, indeterminant in growth habit, and/or extremely late in maturity, were discarded in order to eliminate confounding effects of these factors on disease evaluation, QTL mapping, and future breeding research. The remaining 145 plants (referred to as the BC(1) population) were genotyped for 141 restriction fragment length polymorphism (RFLP) markers and 23 resistance gene analogs (RGAs), and a genetic linkage map was constructed. BC(1) plants were evaluated for disease symptoms throughout the season, and the area under the disease progress curve (AUDPC) and the final percent defoliation (disease severity) were determined for each plant. BC(1) plants were self-pollinated and produced BC(1)S(1) seed. The BC(1)S(1) population, consisting of 145 BC(1)S(1) families, was grown and evaluated for disease symptoms in replicated field trials in two subsequent years (1999 and 2000) and AUDPC and/or final percent defoliation were determined for each family in each year. Two QTL mapping approaches, simple interval mapping (SIM) and composite interval mapping (CIM), were used to identify QTLs for EB resistance in the BC(1) and BC(1)S(1) populations. QTL results were highly consistent across generations, years and mapping approaches. Approximately ten significant QTLs (LOD >/= 2.4, P 57% of the total phenotypic variation. All QTLs had the positive alleles from the disease-resistant parent. The good agreement between results of the BC(1) and 2 years of the BC(1)S(1) generations indicated the stability of the identified QTLs and their potential usefulness for improving tomato EB resistance using marker-assisted selection (MAS). Further inspections using SIM and CIM indicated that six of the ten QTLs had independent additive effects and together could account for up to 56.4% of the total phenotypic variation. These complementary QTLs, which were identified in two generations and 3 years, should be the most useful QTLs for MAS and improvement of tomato EB resistance using PI126445 as a gene resource. Furthermore, the chromosomal locations of 10 of the 23 RGAs coincided with the locations of three QTLs, suggesting possible involvement of these RGAs with EB resistance and a potential for identifying and cloning genes which confer EB resistance in tomato. PMID- 12582600 TI - Production of asymmetric hybrids between Arabidopsis thaliana and Brassica napus utilizing an efficient protoplast culture system. AB - Application of the protoplast culture method developed for Brassica protoplasts to protoplasts of Arabidopsis thaliana has increased the opportunities for interspecific hybridizations involving Arabidopsis. A more-efficient and much simpler method was established compared to the earlier-reported protocol developed for A. thaliana protoplasts in which alginate beads were utilized. Mesophyll protoplasts of A. thaliana (ecotypes 'Landsberg erecta' and 'Wassilewskija') were cultured in the modified 8p liquid medium, which had been developed for Brassica protoplasts. For comparison, protoplasts were cultured in sodium alginate beads supplied with B5 medium according to the protocol for A. thaliana. The protoplasts divided with high frequencies in the 8p medium, and calli proliferated more rapidly than in the sodium alginate beads. High frequencies of shoot differentiation and regeneration were observed in calli of both ecotypes, from about 30% in the ecotype 'Wassilewskija' to about 60% for 'Landsberg erecta'. The more-rapidly the calli developed, the higher the regeneration frequencies were. Asymmetric hybrids between A. thaliana and Brassica napus were obtained by treating the protoplasts of A. thaliana with iodoacetamide (IOA) and B. napus protoplasts with UV-irradiation before fusion with polyethylene glycol (PEG). By using the culture procedure developed for Brassica protoplasts, calli developed and plants were regenerated. Although most of the plants regenerated after cell fusion were A. thaliana-like and were judged to be escapes from IOA treatment, more than ten plants showed hybrid features of both morphological and molecular characters. Among the hybrids that have flowered so far, both male-fertile and male-sterile plants have been obtained. Back crossings to A. thaliana are now in progress as is morphological and molecular characterization of the plants. PMID- 12582601 TI - Comparative analysis of QTLs affecting domestication traits between two domesticated x wild pearl millet ( Pennisetum glaucum L., Poaceae) crosses. AB - Comparative mapping of Quantitative trait loci (QTLs) involved in domestication of adaptative syndrome traits of pearl millet was realized at the intra-specific level using two F(2) populations derived from domesticated ( Pennisetum glaucum ssp. glaucum) x wild ( Pennisetum glaucum ssp. monodii) crosses. The two domesticated parents analyzed differ in their geographical origins, agronomic characteristics and life cycles. In both populations, two regions of the genome were identified on linkage groups 6 and 7, that controlled most of the key morphological differences. The importance of these two linkage groups reveals their central role both in the developmental control of spikelet structure and in the domestication process of this crop. In contrast, QTLs involved in traits that are components of yield and measure differences in resource allocation (such as the shape of the spike, the number of spikes per plant and plant height) show a low level of correspondence among our two crosses. The results of the comparative mapping between cereals, although preliminary, reveal that genes involved in seed shattering could correspond in maize, rice and sorghum. The evolutionary significance of our results, and especially the relationships between genome organization and cereal domestication, are discussed. The potential use of these results in pearl millet genetic-resources enhancement are presented. PMID- 12582602 TI - Different maternal origins of Japanese lowland and upland rice populations. AB - Plastid subtype ID (PS-ID) sequences were determined from sequence data based on CA repeats between genes rpl16 and rpl14 in Japanese lowland and upland cultivars. The PS-ID sequences of Japanese rice cultivars showed that there are different maternal origins between lowland and upland cultivars. One subtype, 6C7A, of PS-ID sequences was predominant in all but one Japanese lowland cultivar and carried a combination of the indica-specific subtype 8C8A and japonica specific nuclear markers for the isozyme genotype. It is probably a nuclear cytoplasmic recombinant resulting from natural out-crossing and succeeding self pollination. The origin of the plastid was re-confirmed by the existence of an indica-specific deletion in the plastid genome. In contrast, the Japanese upland cultivars showed two subtypes, 7C6A and 6C7A, of PS-ID sequences. An upland specific isozyme allele as a nuclear marker was equally predominant in cultivars carrying each subtype. The existence of these particular upland-specific nuclear and cytoplasmic genotypes suggests that the origin of Japanese upland cultivars is different from that of Japanese lowland cultivars. Cultivars carrying the upland-specific nuclear genotype are common in Southeast Asia, but the combination of the upland-specific nuclear and cytoplasmic genotypes which is the same as the Japanese upland predominant type was found in cultivars only in Taiwan and Indonesia. Japanese upland cultivars are closely related to those cultivars. PMID- 12582603 TI - Mapping quantitative trait loci controlling seed longevity in rice ( Oryza sativa L.). AB - Quantitative trait loci (QTLs) controlling seed longevity in rice were identified using 98 backcross inbred lines (BILs) derived from a cross between a japonica variety Nipponbare and an indica variety Kasalath. Seeds of each BIL were kept for 12 months at 30 degrees C in dry conditions to promote loss of viability. To measure seed longevity, we performed an additional aging-processing treatment for 2 months at 30 degrees C maintaining seeds at 15% moisture content. We measured the germination percent of these treated seeds at 25 degrees C for 7 days as the degree of seed longevity. The germination of BILs ranged from 0 to 100% with continuous variation. Three putative QTLs for seed longevity, qLG-2, qLG-4 and qLG-9, were detected on chromosome 2, 4 and 9, respectively. Kasalath alleles increased the seed longevity at these QTLs. The QTL with the largest effect, qLG 9, explained 59.5% of total phenotypic variation in BILs. The other two QTLs, qLG 2 and qLG-4, explained 13.4 and 11.6% of the total phenotypic variation, respectively. We also verified the effect of the Kasalath allele of qLG-9 using chromosome segment substitution lines. Furthermore, QTLs for seed dormancy were identified on chromosomes 1, 3, 5, 7 and 11. Based on the comparison of the chromosomal location of QTLs for seed longevity and seed dormancy, these traits seem to be controlled by different genetic factors. PMID- 12582604 TI - Megagametophyte-derived linkage maps of white spruce ( Picea glauca) based on RAPD, SCAR and ESTP markers. AB - We have constructed linkage maps for two parents of white spruce [ Picea glauca (Moench) Voss]. Haploid megagametophytes from 92 and 96 seeds of parents M2 and 80132, respectively, were analysed with RAPD, SCAR and ESTP markers. Fragments segregating in a 1:1 Mendelian ratio were classified and mapped using MAPMAKER, GMENDEL and JOINMAP. For M2, the analysis with JOINMAP resulted in 165 loci (152 RAPDs, 3 SCARs and 10 ESTPs) mapping to 23 linkage groups and covering 2,059.4 cM(Kosambi function, K). For 80132, the analysis resulted in 144 loci (137 RAPDs, 1 SCAR and 7 ESTPs) mapping to 19 linkage groups and covering 2,007.7 cM(K). The maps covered 87 and 73% of the entire genome of parents M2 and 80132, respectively. Similar results were obtained with MAPMAKER and GMENDEL. A comparison was made between the two individual maps and 16 loci were shared between the two maps. PMID- 12582605 TI - Differentiation of Japanese green tea cultivars as revealed by RFLP analysis of phenylalanine ammonia-lyase DNA. AB - Japanese green tea cultivars and 463 local tea plants including mountainous tea, yama-cha, were analyzed to determine the process of differentiation of Japanese tea plants using phenylalanine ammonia lyase (PAL) as a DNA marker. The main DNA fragments detected by RFLP analysis, which were named A, B and D, were inherited as multiple allelic genes at one locus. Japanese tea cultivars were divided into five groups according to RFLPs: AA, AB, AD, BD and DD. The AA group included many cultivars selected from local tea plants. The BD group consisted of cv Yabukita or descendants from Yabukita produced by artificial crossing. There was no BB group of cultivars. Allelic frequencies of A, B and D were 0.66, 0.08 and 0.22, respectively, and these values were same in tea plants collected from all regions of Japan. Since the frequencies in yama-cha and local tea plants were also the same, it is thought that these teas have the same origin. These results indicate a process of differentiation from the ancestral material presumably introduced from China to the local tea plants and, finally, cultivars which were produced by selecting from local tea plants and crossing. PMID- 12582606 TI - Genes responding to vernalization in hexaploid wheat. AB - Genotype-specific gene expression in response to vernalization in common wheat was examined by the differential display method. Two near-isogenic lines of Vrn A1 ( Vrn-A1 for the spring type and vrn-A1 for the winter type) were treated by vernalization of developing embryos in detached-ear cultures. This treatment was effective to promote vrn-A1 genotypes to head at a time equivalent to that of Vrn A1. Differential cDNA fragments were isolated by the RT-PCR method from embryos subjected to vernalization treatments for 2- and 4-weeks at DAP10 and DAP20 stages, respectively. Among 110 differential cDNA fragments isolated, 48 were examined for their chromosomal locations and designated as wec ( wheat- embryo cold treatment) genes. Seven wec genes showed genotype-specific expression in response to vernalization. The statistical analysis utilizing two recombinant inbred lines showed that four wec genes were significantly associated with heading factors. PMID- 12582607 TI - Assignment of oat linkage groups to microdissected Avena strigosa chromosomes. AB - Microdissection of metaphase chromosome preparations of diploid oat Avena strigosa (2n = 14) allowed isolation of the three individual chromosomes with distinct morphologies, numbers 2, 3 and 7. Using a PCR approach based on the DNA of microdissected chromosomes, STS derivatives of RFLP markers, genetically mapped in Avena spp. linkage maps, have been physically assigned to these three chromosomes. Based on either two or four RFLP-derived STS markers, the A. strigosa chromosomes 2 and 3 were found to be homoeologous to the oat linkage groups C and E, respectively. With the DNA of chromosome 7, four RFLP-derived STS markers located within the central part of linkage group F and two distal ends of linkage group G were amplified. Accordingly, chromosome 7 corresponds to linkage group F and, most probably, is involved in an A. strigosa-specific chromosomal translocation relative to the diploid species Avena atlantica and Avena hirtula, of which the cross progeny was used for linkage mapping of the tested RFLP clones. PMID- 12582608 TI - A reference map of Cucumis melo based on two recombinant inbred line populations. AB - A composite genetic melon map was generated based on two recombinant inbred line (RI) populations. By analyzing the segregation of 346 AFLPs, 113 IMAs and phenotypic characters on a RI population of 163 individuals derived from the cross Vedrantais x PI 161375, a first map was constructed. About 20% of the molecular markers were skewed, and the residual heterozygosity was estimated at 4.43% which was not significantly different from the theoretical value of 4.2%. The genome distribution of molecular markers among the 12 linkage groups was not different from a random distribution with the exception of linkage group XII which was found significantly less populated. The genome distributions of IMAs and AFLPs were complementary. AFLPs were found mainly in the middle of each linkage group and sometimes clustered, whereas IMAs were found mainly at the end. A total of 318 molecular markers, mainly AFLP and IMA markers, were mapped on 63 RIs of the second population, Vedrantais x PI 414723. Comparison of the maps enables one to conclude that AFLPs and IMAs of like molecular size, amplified with the same primer combination, correspond to the same genetic locus. Both maps were joined through 116 common markers comprising 106 comigrating AFLPs/IMAs, plus five SSRs and five phenotypic markers. The integrated melon map contained 668 loci issuing from the segregation of 1,093 molecular markers in the two RI populations. The composite map spanned 1,654 cM on 12 linkage groups which is the haploid number of chromosomes in melon. Thirty two known-function probes, i.e. known-function genes (9) and morphological traits (23), were included in this map. In addition, the composite map was anchored to previously published maps through SSRs, RFLPs and phenotypic characters. PMID- 12582609 TI - Efficient development of dinucleotide microsatellite markers in Norway spruce ( Picea abies Karst.) through dot-blot selection. AB - The development of microsatellite markers can be a time-consuming process, especially in species such as conifers where many microsatellites have been shown to be associated with the repetitive fraction of the genome and to produce complex banding patterns following electrophoresis. Therefore, procedures to eliminate this fraction from further processing are sought. In this paper, we report on the development of 53 dinucleotide SSR markers in Norway spruce, 35 of which (66%) produce simple, polymorphic patterns. This high efficiency is obtained by introducing a dot-blot selection against high copy number sequences, performed on the microsatellite-containing clones. The resulting markers turned out to be polymorphic and useful for population genetic studies and for linkage mapping. Seven additional markers that were not subject to the dot-blot selection are also presented. PMID- 12582610 TI - Identification of microsatellite markers linked to Russian wheat aphid resistance genes Dn4 and Dn6. AB - The Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), is a serious economic pest of wheat worldwide. Host plant resistance is the preferred method to control RWA infestations. The identification and mapping of RWA-resistant genes and the development of resistant wheat cultivars can be facilitated through the use of molecular markers. In the present study, microsatellite (SSR) markers linked to the RWA-resistant genes Dn4 and Dn6 were identified using several F(2) mapping populations derived from crosses of susceptible wheat cultivars and resistant sources. Two flanking microsatellite markers Xgwm106 and Xgwm337 are linked in coupling phase with Dn4 on the short arm of wheat chromosome 1D at 7.4 cM and 12.9 cM, respectively. Two other microsatellite markers Xgwm44 and Xgwm111 are linked to Dn6 in coupling phase near the centromere on the short arm of chromosome 7D at 14.6 cM and 3.0 cM, respectively. This is the first report on the chromosomal location of Dn6, which proved to be either allelic or tightly linked to Dn1, Dn2 and Dn5. This result of Dn6 location contradicts previous reports that Dn6 was independent of Dn1, Dn2 and Dn5. The linked markers can be conveniently used for marker-assisted selection in wheat breeding programs for the identification and/or pyramiding of Dn4 and Dn6 genes. PMID- 12582611 TI - Identification and characterization of DNA markers associated with a locus conferring virulence on barley in the plant pathogenic fungus Cochliobolus sativus. AB - Cochliobolus sativus is a plant pathogenic fungus that causes spot blotch on barley and wheat. Virulence of a pathotype-2 isolate (ND90Pr) on barley cultivar Bowman was previously determined to be controlled by a single locus. To identify DNA markers associated with this virulence locus, amplified fragment length polymorphism (AFLP) analysis was conducted on 104 progeny isolates derived from a cross between isolates ND90Pr (exhibiting high virulence on Bowman) and ND93-1 (exhibiting low virulence on Bowman). Among 115 AFLP markers identified, 14 were linked to the virulence locus VHv1 in isolate ND90Pr, six of which co-segregated with VHv1. Two (E-AG/M-CA-207 and E-AG/M-CG-121) of the six co-segregating AFLP markers were cloned and used to probe genomic DNAs from the fungal parents and progeny. Both markers hybridized only with DNAs from ND90Pr and the virulent progeny. These two cloned markers were also used as probes to survey field isolates of C. sativus collected from different regions of the world and again only hybridized to DNAs from isolates that had the same virulence phenotype as ND90Pr. The results of this study indicate that E-AG/M-CA-207 and E-AG/M-CG-121 are closely linked to VHv1 and are unique to isolates carrying the virulence locus. Development of a linkage group, coupled with the identification of closely linked molecular markers, will facilitate the cloning of the virulence gene VHv1 in C. sativus by map-based cloning. PMID- 12582612 TI - Resistance gene homologues in melon are linked to genetic loci conferring disease and pest resistance. AB - Genomic and cDNA fragments with homology to known disease resistance genes (RGH fragments) were cloned from Cucumis melo using degenerate-primer PCR. Fifteen homologues of the NBS-LRR gene family have been isolated. The NBS-LRR homologues show high divergence and, based on the partial NBS-fragment sequences, appear to include members of the two major subfamilies that have been described in dicot plants, one that possesses a TIR-protein element and one that lacks such a domain. Genomic organization of these sequences was explored by DNA gel-blot analysis, and conservation among other Cucurbitaceae was assessed. Two mapping populations that segregate for several disease and pest resistance loci were used to map the RGH probes onto the melon genetic map. Several NBS-LRR related sequences mapped to the vicinity of genetic loci that control resistance to papaya ringspot virus, Fusarium oxysporum race 1, F. oxysporum race 2 and to the insect pest Aphis gossypii. The utility of such markers for breeding resistant melon cultivars and for cloning the respective R-genes is discussed. PMID- 12582613 TI - Comparison of ISSR and RAPD markers to characterize three Chilean Nothofagus species. AB - The present study is the first report of fingerprinting on three Chilean Nothofagus species using ISSR and RAPD markers; 61 Nothofagus nervosa, 32 Nothofagus obliqua and 32 Nothafagus dombeyi individual trees, sampled from collection and natural sites, were analyzed. Among 45 primers tested, the 6 ISSR and 6 RAPD primers selected for the analysis generated a total of 63 ISSR and 42 RAPD fragments. A high proportion of polymorphic bands, ranging from 97% and 80%, was found using both markers. A similar number of private and marker bands was generated by both markers in all the species examined and one discriminant ISSR fragment was obtained for N. dombeyi. Jaccard and Dice similarity indices were used to evaluate pairwise genetic divergence; cluster analysis of the similarity matrices was performed to estimate the intra- and inter-specific genetic diversity, and PCA analysis was employed to evaluate the resolving power of the markers to differentiate between the species. These analyses, carried out for both markers, allowed us to identify three main groups corresponding to the three Nothofagus species. The results of the present study can be seen as a starting point for future researches on the population and evolutionary genetics of these species. PMID- 12582614 TI - Molecular cloning of the wheat CK2alpha gene and detection of its linkage with Vrn-A1 on chromosome 5A. AB - The casein kinase CK2 is one of the major multifunctional protein kinases in cells that is expressed ubiquitously and is essential for survival. The alpha subunit of CK2 is thought to be involved in light-regulated gene expression and rhythmic expression of genes by circadian rhythm in plants. The rice chromosome-3 region containing the photoperiod-response Hd6 gene, an orthologue of the CK2alpha genes of Arabidopsis and maize, is in synteny with the wheat chromosome 5A Vrn-A1 region. This evidence proposes two possibilities, first the wheat Vrn A1 is an orthologue of the rice CK2alpha, and second the wheat CK2alpha which has not yet been identified is located independently but tightly linked to Vrn-A1. To clarify whether the wheat CK2alpha gene is conserved in the Vrn-A1 region and to elucidate the above two possibilities, we attempted to isolate this gene from the wheat cDNA library and to map it on the chromosome-5A region that is syntenous to the rice Hd6 region. The isolated cDNA clone showed an extremely high homology with the Arabidopsis CK2alpha gene. Using this clone as a probe genomic Southern blot analyses of the aneuploid lines available in Chinese Spring assigned the wheat homologue of CK2alpha to the long arm of chromosome 5A. Furthermore, a linkage analysis using an F(2) population having recombination in the Vrn-A1 region revealed that the wheat CK2alpha, designated as tck2a, is tightly linked to Vrn-A1 by 1.1 cM PMID- 12582615 TI - Comparison of homoeologous group-6 short arm physical maps of wheat and barley reveals a similar distribution of recombinogenic and gene-rich regions. AB - Eighty two new loci, mapped with 51 DNA clones, were added to the earlier deletion maps of the homoeologous group-6 short arms of hexaploid wheat ( Triticum aestivum L. em Thell., 2n = 6 x = 42, AABBDD). There are now 41, 56 and 52 loci mapped on deletion maps of 6AS, 6BS and 6DS, respectively. The linear order of orthologous loci in all three arms appears to be identical. The majority of the loci are located in the distal one-half of the three arms. There seems to be an increased marker/gene density from the centromeric to the telomeric regions in each arm, and the marker density in comparable physical regions is similar on all three maps. Recombination is not uniformly distributed along the chromosome arms; 60% of recombination occurs in the distal one-third of each arm. Recombination increases from the proximal region to the distal end in a nonlinear pattern. The distribution of loci and recombination along each of the three chromosome arms is highly correlated. Comparison of the 6BS deletion map from this study and a 6HS physical map of barley ( Hordeum vulgare L., 2n = 2 x = 14, HH) reveals a remarkably similar distribution of recombinogenic and gene-rich regions between the two chromosome arms, suggesting that the distribution patterns of genes may be conserved in the homoeologous group-6 chromosome short arms of wheat and barley. A consensus map of wheat group-6 short arms containing 46 orthologous loci was constructed. Comparison of the consensus map with published linkage maps of Triticeae group-6 chromosome short arms indicates that the linear order of the loci on the maps has been largely conserved. Evidence from this study does not support the existence of a 2BS-6BS reciprocal terminal translocation. PMID- 12582616 TI - Analysis of the duplicated CHS1 gene related to the suppression of the seed coat pigmentation in yellow soybeans. AB - Seed coat color in soybean is controlled by the classically defined I ( Inhibitor) locus. The seeds of most commercial soybean varieties are yellow due to the presence of a dominant allele of the I locus ( I: yellow seed coat, or i(i) : pigmented hilum and yellow seed coat), which inhibits seed coat pigmentation. Analysis of spontaneous mutations from I (yellow seed coat) to i (pigmented seed coat) has shown that these mutations are correlated with the deletion of a duplicated chalcone synthase gene-1 ( CHS1) region. In the current study, we isolated the duplicated CHS1 region from a soybean cultivar with a I/I genotype (cv Miyagi shirome) and determined its structure. The results showed that the duplicated CHS1 contained intact regulatory and coding regions. We designated the duplicated CHS1 as ICHS1. In the hypocotyls of Miyagi shirome, the cDNA derived from ICHS1 mRNA was identified by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, whereas in the immature seed coats it was suggested that the amount of transcripts from ICHS1 and/or another type of CHS1 ( CHS1.1) was very low. Interestingly, in the Miyagi shirome genome with a I/I genotype, ICHS1 was closely linked to the truncated CHS3, and sequence comparison showed that this cluster probably arose from the CHS1-CHS3 cluster by a 1.8-kb deletion event. PMID- 12582617 TI - Isolation and characterization of microsatellites in Brassica rapa L. AB - We report here the isolation and characterization of microsatellites, or simple sequence repeats (SSRs), in Brassica rapa. The size-fractionated genomic library was screened with (GA)(15) and (GT)(15) oligonucleotide probes. A total of 58 clones were identified as having the microsatellite repeats, and specific primer pairs were designed for 38 microsatellite loci. All primer pairs, except two, amplified fragments having the sizes expected from the sequences. Of the 36 primer pairs, 35 amplified polymorphic loci in 19 cultivars of B. rapa, while monomorphism was observed in only one primer pair. A total of 232 alleles was identified by the 36 primer pairs in 19 cultivars of B. rapa, and these primer pairs were examined also in nine Brassicaceae species. Most of the 36 primer pairs amplified the loci in the Brassicaceae species. Segregation of the microsatellites was studied in an F(2) population from a cross of doubled-haploid lines DH27 x G309. The microsatellites segregated in a co-dominant manner. These results indicate that the microsatellites isolated in this study were highly informative and could be useful tools for genetic analysis in B. rapa and other related species. PMID- 12582618 TI - Studies on the origin and evolution of tetraploid wheats based on the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA. AB - In this study, the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA in the tetraploid wheats, Triticum turgidum (AABB) and Triticum timopheevii (AAGG), their possible diploid donors, i.e., Triticum monococcum (AA), Triticum urartu (AA), and five species in Aegilops sect. Sitopsis (SS genome), and a related species Aegilops tauschii were cloned and sequenced. ITS1 and ITS2 regions of 24 clones from the above species were compared. Phylogenetic analysis demonstrated that Aegilops speltoides was distinct from other species in Aegilops sect. Sitopsis and was the most-likely donor of the B and G genomes to tetraploid wheats. Two types of ITS repeats were cloned from Triticum turgidum ssp. dicoccoides, one markedly similar to that from T. monococcum ssp. boeoticum (AA), and the other to that from Ae. speltoides (SS). The former might have resulted from a recent integression event. The results also indicated that T. turgidum and T. timopheevii might have simultaneously originated from a common ancestral tetraploid species or be derived from two hybridization events but within a very short interval time. ITS paralogues in tetraploid wheats have not been uniformly homogenized by concerted evolution, and high heterogeneity has been found among repeats within individuals of tetraploid wheats. In some tetraploid wheats, the observed heterogeneity originated from the same genome (B or G). Three kinds of ITS repeats from the G genome of an individual of T. timopheevii ssp. araraticum were more divergent than that from inter-specific taxa. This study also demonstrated that hybridization and polyploidization might accelerate the evolution rate of ITS repeats in tetraploid wheats. PMID- 12582619 TI - Mapping QTLs for sucrose content, yield and quality in a sugar beet population fingerprinted by EST-related markers. AB - Seventy five expressed sequence tags (ESTs) that are associated with functions in carbohydrate and nitrogen metabolism were genotyped in 108 plants of an F2 population of sugar beet ( Beta vulgaris L.) segregating for sugar quality and yield parameters. Supplemented by known RFLP and AFLP markers, the resulting map spans 446 cM of the 758-Mbp genome of sugar beet. F3 test-cross plants were analysed for corrected sugar yield, beet yield, ion balance and the content of sugar, amino nitrogen, potassium and sodium in six locations. Twenty one significant quantitative trait loci (QTLs) were detected using the composite interval mapping approach. Expressed genes flanking the QTLs were identified in all cases. Correlations between QTLs and potential candidate genes are discussed. PMID- 12582620 TI - SNP markers for black-grass ( Alopecurus myosuroides Huds.) genotypes resistant to acetyl CoA-carboxylase inhibiting herbicides. AB - Chloroplastic acetyl CoA-carboxylase (ACCase) is the target of widely used, specific graminicide herbicides: cyclohexanediones (CHDs) and aryloxyphenoxypropionates (APPs). Resistance to these compounds is a worldwide, increasing problem. Population genetic studies aimed at understanding the dynamics of this situation and the diffusion of resistance genes within and between weed populations are challenging because biological assays are not adequate for this purpose, and because different mechanisms of resistance confer a similar resistance phenotype. Molecular markers for specifically detecting resistance genes are therefore urgently needed to conduct such studies. For this purpose, we cloned and sequenced the whole gene encoding chloroplastic ACCase in Alopecurus myosuroides Huds. (Black-grass). We identified two point mutations at nucleotide 5,341 that both cause an isoleucine-leucine substitution at position 1,781. Three bi-directional allele-specific PCR assays were developed, each detecting two distinct ACCase alleles with a single PCR reaction. The sensitivity of 1,190 seedlings of A. myosuroides to one CHD and one APP was determined. Genotyping revealed that, although resistant plants were only selected by APPs, the (1,781)Leu ACCase allele is a widespread, dominant gene of resistance to both APPs and CHDs. No other ACCase allele associated with resistance could be identified in this work. Useful applications of allele-specific PCR markers are population genetic studies as well as routine molecular diagnosis of herbicide resistance. PMID- 12582621 TI - Brassica napus DNA markers linked to white rust resistance in Brassica juncea. AB - White rust, caused by Albugo candida, is an economically important disease of Brassica juncea mustard. The most efficient and cost effective way of protecting mustard plants from white rust is through genetic resistance. The development of canola quality B. juncea through interspecific crosses of B. juncea with Brassica napus has lead to the introgression of white rust resistance from B. napus into B. juncea. The objective of this study was to identify DNA markers for white rust resistance, derived from the introgressed B. napus chromosome segment, in a BC(3)F(2) population of condiment B. juncea mustard. This segregating population was phenotyped for white rust reaction and used to screen for AFLP markers associated with white rust resistance using bulked segregant analysis. Segregation data indicated that a single dominant gene controlled resistance to white rust. Eight AFLP markers linked to white rust resistance were identified, all derived from B. napus. The B. napus chromosome segment, carrying the white rust resistance gene ( Ac2V(1)), appeared to have recombined with the B. juncea DNA since recombinant individuals were identified. Comparative mapping of the eight B. napus-derived AFLP markers in a typical B. napus mapping population was inconclusive; therefore, the size of the introgressed B. napus fragment could not be determined. PMID- 12582622 TI - A soybean gene encoding a proline-rich protein is regulated by salicylic acid, an endogenous circadian rhythm and by various stresses. AB - A cDNA clone of a soybean gene encoding a proline-rich protein (PRP) was characterized and designated SbPRP ( Soy bean Proline- rich Protein). The SbPRP protein is a putative bimodular protein of 126 amino acids with a proline-rich domain and a hydrophobic cysteine-rich domain plus a signal peptide at the N terminal. Southern analysis indicates the presence of a single copy of the SbPRP gene in the soybean genome. The SbPRP gene expression was investigated and the results demonstrate that it accumulates in leaves and epicotyls of soybean seedlings, but not in cotyledons, hypocotyls and roots. The SbPRP mRNA was also expressed in response to salicylic acid and virus infection. In addition, the SbPRP gene transcription was regulated by circadian rhythm, salt stress, drought stress and plant hormones. These results indicate that the SbPRP gene might play a role in plant responses to multiple internal and external factors. PMID- 12582623 TI - Genomic organization of glycinin genes in soybean. AB - Glycinin is the predominant seed storage protein in most soybean varieties. Previously, five major genes (designated Gy1 to Gy5) encoding glycinin subunits have been described. In this report two new genes are identified and mapped: a glycinin pseudogene, gy6, and a functional gene, Gy7. Messenger RNA for the gy6 pseudogene is not detected in developing seeds. While Gy7 mRNA was present at the midmaturation stage of seed development in the soybean variety Resnik, the steady state amount of this message was at least an order of magnitude less-prevalent than the mRNA encoding each of the other five glycinin subunits. Even though the amino-acid sequence of the glycinin subunit G7 is related to the other five soybean 11S subunits, it does not fit into either the Group-1 (G1, G2, G3) or the Group-2 (G4, G5) glycinin subunit families. The Gy7 gene is tandemly linked 3' to Gy3 on Linkage Group L (chromosome 19) of the public molecular linkage map. By contrast, the gy6 gene occupies a locus downstream from Gy2 on Linkage Group N (chromosome 3) in a region that is related to the position where Gy7 is located on chromosome 19. PMID- 12582624 TI - Trans-activation of the maize transposable element, Ds, in Brassica napus. AB - A two-component transposable element system consisting of a stabilized Activator ( Ac(st)) and a chimeric Dissociation ( Ds) element has been introduced into the genome of Brassica napus. Analyses performed on F2 progeny derived from crosses between Ac(st)- and Ds-bearing parents confirm that Ac transposase catalyzes the somatic excision of the Ds element in both embryonic and non-embryonic tissues of this important crop species. The data further reveal that the vast majority of plants containing both Ac(st) and Ds exhibit Ds excision. However, the level of excision is low and germinal Ds excision events are not observed. We estimate that germinal excision of Ds occurs at a frequency of < 0.2%. RT-PCR analysis of the Ac(st) transcript in somatically active seedlings reveals that introns III and IV are highly misprocessed. The pattern of transcript processing is very similar to that observed in germinally inactive but somatically active Arabidopsis seedlings. We suggest that Ds excision activity in B. napus is highly dependent on the efficiency of Ac(st) transcript processing. PMID- 12582625 TI - A high degree of genetic diversity is revealed in Isatis spp. (dyer's woad) by amplified fragment length polymorphism (AFLP). AB - Genetic diversity in 38 genotypes, representing 28 individual genotypes from five landraces of Isatis tinctoria (three German: Tubingen, Potsdam and Erfurt, one Swiss and one English), five genotypes of Isatis indigotica (Chinese woad) and five genotypes of Isatis glauca, were investigated using AFLP analysis. Five primer combinations detected a total of 502 fragments of which 436 (86.9%) were polymorphic. The level of polymorphism recorded within each species was 29.8, 86.9 and 35.8% for I. indigotica, I. tinctoria and I. glauca, respectively. Clearly, genetic diversity within I. tinctoria was greater than that observed in I. indigotica or I. glauca. Cluster analyses of the AFLP data using UPGMA and PCO revealed the complete separation of the genotypes of each species into distinct groups. I. indigotica separated as an entirely independent group, whereas I. glauca formed a separate cluster within the I. tinctoria group. Indeed, I. tinctoria and I. glauca are more closely related to each other than either is to I. indigotica. In addition, the genotypes of each landrace, apart from one from the English group, were clearly discriminated. However, the anomalous genotype did associate with the rest of its group when it was linked with the Erfurt group. These results provide new and useful information about the make-up of the Isatis genome, which has not previously been evaluated. They will be useful in the selection of plant material for variety development and conservation of the gene-pool. PMID- 12582626 TI - On chloroplast DNA variations in the olive ( Olea europaea L.) complex: comparison of RFLP and PCR polymorphisms. AB - Previous papers have dealt with olive chloroplastic DNA (cpDNA) variation revealed using several methods (RFLPs, PCR-RFLPs and microsatellites) and have led to different conclusions. This paper aims to reconsider these divergences. A Southern approach was applied to reveal polymorphism. We used chloroplast DNA of Phillyrea media as a probe. Based on these data, only four chlorotypes were identified in the olive complex. The number of detected lineages was lower than reported in the literature using a direct cpDNA RFLP approach, and was insufficient to distinguish the North African subspecies europaea, maroccana, guanchica and laperrinei. Furthermore, one individual considered belonging to the subspecies laperrinei was questionable. Using other cpDNA and mitochondrial DNA (mtDNA) polymorphisms - based on PCR and RFLP methods, respectively - we showed that this individual displays the cytoplasmic lineage CE1-ME1 characteristic of most Eastern mediterranean cultivars and of Olea europaea subsp. laperrinei from Hoggar. However, based on RAPDs, this individual appeared as mislabelled and probably corresponded to a Mediterranean cultivar or a feral form. In addition, we checked O. e. subsp. laperrinei herbarium samples using two cpDNA microsatellites, which revealed polymorphisms. These also supported that both populations from Niger and Algeria displayed a chlorotype related to CE1. Consequently, based on cpDNA, the relationships of O. e. subsp. laperrinei from Hoggar with a Mediterranean lineage appeared well supported, whereas the South West Moroccan and Macaronesian olives appeared in a different clade using both mtDNA and cpDNA polymorphisms. We conclude that methods based on PCR reveal more polymorphisms in the cpDNA and lead to more-reliable results that the classical RFLP method. PMID- 12582627 TI - Characterisation of polymorphic microsatellite markers from Aegilops tauschii and transferability to the D-genome of bread wheat. AB - Microsatellites were isolated from a Aegilops tauschii (the D-genome donor of bread wheat) library enriched for various motifs. Primers generated from the flanking region of the microsatellites were used successfully to amplify the corresponding loci in the D genome of bread wheat. Additional amplification sometimes also occurred from the A and B genomes. The majority of the microsatellites contained (GA)(n) and (GT)(n) motifs. GA and GT repeats appeared to be both more abundant in this library and more polymorphic than other types of repeats. The allele number for both types of dinucleotide repeats fitted a Poisson distribution. Deviance analysis showed that GA and GT were more polymorphic than other motifs in bread wheat. Within each motif type (di-, tri- and tetra-nucleotide repeats), repeat number has no influence on polymorphism. The microsatellites were mapped using the Triticum aestivum Courtot x Chinese Spring mapping population. A total of 100 markers was developed on this intraspecific map, mainly on the D genome. For polyploid species, isolation of microsatellites from an ancestral diploid donor seems to be an efficient way of developing markers for the corresponding genome in the polyploid plant. PMID- 12582628 TI - Bay-0 x Shahdara recombinant inbred line population: a powerful tool for the genetic dissection of complex traits in Arabidopsis. AB - Natural genetic variation in Arabidopsis is considerable, but has not yet been used extensively as a source of variants to identify new genes of interest. From the cross between two genetically distant ecotypes, Bay-0 and Shahdara, we generated a Recombinant Inbred Line (RIL) population dedicated to Quantitative Trait Locus (QTL) mapping. A set of 38 physically anchored microsatellite markers was created to construct a robust genetic map from the 420 F6 lines. These markers, evenly distributed throughout the five chromosomes, revealed a remarkable equilibrium in the segregation of parental alleles in the genome. As a model character, we have analysed the genetic basis of variation in flowering time in two different environments. The simultaneous mapping of both large- and small-effect QTLs responsible for this variation explained 90% of the total genotypic variance. Two of the detected QTLs colocalize very precisely with FRIGIDA and FLOWERING LOCUS C genes; we provide information on the polymorphism of genes confirming this hypothesis. Another QTL maps in a region where no QTL had been found previously for this trait. This confirms the accuracy of QTL detection using the Bay-0 x Shahdara RIL population, which constitutes the largest in size available so far in Arabidopsis. As an alternative to mutant analysis, this population represents a powerful tool which is currently being used to undertake the genetic dissection of complex metabolic pathways. PMID- 12582629 TI - Improving linkage analysis in outcrossed forest trees - an example from Acacia mangium. AB - Mapping in forest trees generally relies on outbred pedigrees in which genetic segregation is the result of meiotic recombination from both parents. The currently available mapping packages are not optimal for outcrossed pedigrees as they either cannot order phase-ambiguous data or only use pairwise information when ordering loci within linkage groups. A new package, OUTMAP, has been developed for mapping codominant loci in outcrossed trees. A comparison of maps produced using linkage data from two pedigrees of Acacia mangium Willd demonstrated that the marker orders produced using OUTMAP were consistently of higher likelihood than those produced by JOINMAP. In addition, the maps were produced more efficiently, without the need for recoding data or the detailed investigation of pairwise recombination fractions which was necessary to select the optimal marker order using JOINMAP. Distances between markers often varied from those calculated by JOINMAP, resulting in an increase in the estimated genome length. OUTMAP can be used with all segregation types to determine phase and to calculate the likelihood of alternative marker orders, with a choice of three optimisation methods. PMID- 12582630 TI - QTL mapping and introgression of yield-related traits from Oryza glumaepatula to cultivated rice ( Oryza sativa) using microsatellite markers. AB - Rice ( Oryza sativa) cultivar development currently faces the task of overcoming yield plateaus, which is difficult due to the narrow genetic base of breeding programs. Oryza glumaepatula is a diploid wild relative of cultivated rice, native to Central and South America, and is therefore a potential source of alleles of agronomic importance to rice breeding programs. We studied 11 agronomic traits in BC(2)F(2) families of the interspecific cross Oryza sativa x O. glumaepatula. Transgressive lines which are almost isogenic to the elite recurrent O. sativa parent were identified for most of these traits. Quantitative trait locus (QTL) analysis was performed by single-point and interval mapping using a molecular map based on 157 microsatellite and STS markers. Marker regions accounting for 14.5 to 72.9% of a phenotypic variation trait were identified in 9 of the 12 rice chromosomes. Positive QTL effects from O. glumaepatula were observed in chromosomal regions associated with tillering and panicle-number traits. PMID- 12582631 TI - The effect of a deficiency and a deletion on recombination in chromosome 1BL in wheat. AB - To test two models of chiasma allocation and the distribution of crossing-over in chromosomes, genetic mapping was performed in normal, deletion and deficiency chromosome arms 1BL of wheat, Triticum aestivum L. Shortening of the chromosome arm, either by a deletion of the proximal half of the arm or by a deficiency of the terminal quarter of the arm's length, significantly reduced the frequency of multiple crossovers but did not affect the distribution of the distal, presumably the first, crossover in the arm. In the deficiency chromosome, the recombination rate in the terminal segment was much higher than that in the same segment of the complete arm. This suggests that recombination frequency is not an inherent characteristic of a segment but depends on the segment's position on the centromere-telomere axis. These observations support the classical model of chiasma distribution along the chromosome based on the point of pairing initiation, chromosome length and the positive chiasma interference. The study also demonstrates that the distribution and frequency of recombination in a chromosome segment can be manipulated. Therefore, even the segments with very low recombination frequencies could be saturated with large numbers of crossover events to produce high-density genetic maps. PMID- 12582632 TI - Mitochondrial DNA variation and RAPD mark oleasters, olive and feral olive from Western and Eastern Mediterranean. AB - The study of genetic diversity within the olive-tree (cultivated and wild forms) may be useful to reveal agronomic traits in the wild germplasm and to try to understand the history of the olive-tree domestication. In this way, a study of nuclear and mitochondrial DNAs of cultivated and wild olives from two Corsican and Sardinian Mediterranean islands was performed using RAPD and RFLP markers. Our results show that most of the varieties and most of the oleasters were separated using the UPGMA dendrogram based on the Nei and Li similarity index. Most of oleasters carried either the MOM or MCK mitotype, characteristic of olives in the Western Mediterranean, whereas most of the varieties carried the ME1 mitotype, characteristic of olives in the East Mediterranean. The results indicate that the combination of mitotype and RAPD markers can be used as a powerful tool for differentiating two groups in the wild forms: the Western true oleasters and the feral forms. The true oleasters are characterized by a Western mitotype and a Western RAPD pattern. Feral forms originate either from varieties or from hybridisation between a variety and an oleaster. Consequently, as expected, some of them aggregated with the varieties from which they were derived. The other feral forms are clustered with the oleasters and were detected only by their mitotype determination. This study has also permitted us to differentiate two populations of cultivated olives in Corsica: one with close relationships with Italian varieties (influenced by the East) and one selected from local oleasters probably due to a better local adaptation than foreign varieties. PMID- 12582633 TI - Quantitative trait loci analysis of growth response to varying nitrogen sources in Arabidopsis thaliana. AB - Nitrogen absorption and assimilation is variable among plants as a result of two factors: the source of nitrogen available and the genetic variation among species within the resulting nitrogen pathways. Several genes involved in nitrogen cycling have been identified, yet little is known about the genes that control quantitative responses to different nitrogen sources. With quantitative trait loci (QTL) mapping in Arabidopsis thaliana recombinant inbred lines (Columbia x Landsberg erecta) we have identified chromosomal regions controlling aerial mass, root mass, and root length when plants are grown in nitrate, ammonium, ammonium nitrate, or low nitrogen treatments. A total of 16 QTL ( P < 0.01) were identified among the nitrogen treatments. Most of the QTL were specific to a single treatment. The percentage additive genetic effects of significant QTL were as high as 17%. Five significant QTL corresponded to the locations of candidate genes associated with nitrogen assimilation, while a few QTL corresponded with candidate genes in the developmental pathways. Most QTL were not shared across treatments, suggesting that there is no optimal genotype for all nitrogen sources. PMID- 12582634 TI - Multiple QTL mapping in related plant populations via a pedigree-analysis approach. AB - QTL mapping experiments in plant breeding may involve multiple populations or pedigrees that are related through their ancestors. These known relationships have often been ignored for the sake of statistical analysis, despite their potential increase in power of mapping. We describe here a Bayesian method for QTL mapping in complex plant populations and reported the results from its application to a (previously analysed) potato data set. This Bayesian method was originally developed for human genetics data, and we have proved that it is useful for complex plant populations as well, based on a sensitivity analysis that was performed here. The method accommodates robustness to complex structures in pedigree data, full flexibility in the estimation of the number of QTL across multiple chromosomes, thereby accounting for uncertainties in the transmission of QTL and marker alleles due to incomplete marker information, and the simultaneous inclusion of non-genetic factors affecting the quantitative trait. PMID- 12582635 TI - Transgenic plants of blue grama grass, Bouteloua gracilis (H.B.K.) Lag. ex Steud., from microprojectile bombardment of highly chlorophyllous embryogenic cells. AB - For the first time, the production of transgenic plants of the forage grass blue grama, Bouteloua gracilis [H.B.K.] Lag. ex Steud., is reported. Transgenic plants containing a gus Colon, two colons nptll fusion driven by a double CaMV35S promoter were obtained by microprojectile bombardment of the highly chlorophyllous embryogenic cell line 'TIANSJ98'. Transformed B. gracilis cell lines resisted a lethal concentration of 160 mg/l of kanamycin for at least 8 months. Chlorophyll development and growth rate were used as useful criteria for discriminating transformed from non-transformed clones. Stable integration of the transgene in the blue grama genome was demonstrated by PCR and Southern hybridization analysis. Expression of the NPTll protein in transgenic plants grown under greenhouse conditions was confirmed indirectly by spraying kanamycin (150-250 mg/l) on plant foliage, and directly by ELISA immunological tests. Control plants sprayed with kanamycin showed foliar necrosis and reduced growth (tillering) compared to plants containing the transgene. NPTll was found in transgenic plants in levels ranging between 12.6 and 29.6 ng/mg FW of cells, as determined by ELISA reactions. PMID- 12582636 TI - Genetic dissection of a genomic region for a quantitative trait locus, Hd3, into two loci, Hd3a and Hd3b, controlling heading date in rice. AB - The rice photoperiod sensitivity gene Hd3 was originally detected as a heading date-related quantitative trait locus localized on chromosome 6 of rice. High resolution linkage mapping of Hd3 was performed using a large segregating population derived from advanced backcross progeny between a japonica variety, Nipponbare, and an indica variety, Kasalath. To determine the genotype of Hd3, we employed progeny testing under natural field and short-day conditions. As a result, two tightly linked loci, Hd3a and Hd3b, were identified in the Hd3 region. Nearly-isogenic lines for Hd3a and Hd3b were selected from progeny using marker-assisted selection. The inheritance mode of both Hd3a and Hd3b was found to be additive. Analysis of daylength response in nearly-isogenic lines of Hd3a and Hd3b showed that the Kasalath allele at Hd3a promotes heading under short-day conditions while that at Hd3b causes late heading under long-day and natural field conditions. PMID- 12582637 TI - Detection of 3-hydroxy-3-methylglutaryl-coenzyme A reductase protein Cm-HMGR during fruit development in melon ( Cucumis melo L.). AB - The fruit size of melon ( Cucumis melo L. reticulatus) is determined by the amount of cell proliferation in the pericarp during early fruit development. During this stage, expression and activity of the 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene is required for fruit growth. In this study, we performed a detailed analysis of the correlation between the expression of melon HMGR (Cm-HMGR) protein and cell division in the pericarp. Flow cytometric analysis revealed that the length of the cell division stage was correlated with the fruit size. Western gel blotting and tissue printing illustrated the temporal and spatial accumulation pattern of Cm-HMGR protein during fruit development. The accumulation of Cm-HMGR transiently increased at the beginning of the cell division stage in the pericarp, where active cell division occurred. The amount of Cm-HMGR was correlated with the length of the cell division period. These results strongly suggest that the expression of Cm-HMGR is involved in the determination of melon fruit size by regulating cell division during early fruit development. PMID- 12582638 TI - Investigation of crossover interference in barley ( Hordeum vulgare L.) using the coefficient of coincidence. AB - Interference, the interaction between recombination events, was analysed in seven mapping populations of barley ( Hordeum vulgare L.). The coefficient of coincidence was applied to investigate the type and position of interference within the genome. Interference was analysed with respect to dependence on the recombination fraction, and simulations were used to obtain test statistics which consider the small sample size of 71-150 double haploid lines. In addition to positive interference in intermediate intervals, strong negative interference, i.e. encouraged double recombination, was found in short intervals. The relationship between recombination fraction and interference could not be described with a uniform function, neither for the entire genome nor for individual chromosomes. The analysis of the position of interference within the genome revealed that interference does not act in the same way in the whole genome. Intervals spanning the centromere exhibited significantly higher means for the coefficient of coincidence than intervals within the chromosome arms, especially with regard to small intervals. In general, positive interference was found in the chromosome arms and no or negative interference in the genetically small but physically large centromeric region. PMID- 12582639 TI - Inheritance and segregation of virus and herbicide resistance transgenes in sugarcane. AB - Transgenic sugarcane parents containing multiple copies of herbicide resistance ( bar) and Sorghum mosaic virus (SrMV) resistance ( hut) genes were crossed with non-transgenic sugarcane varieties. Segregation of the transgenes in the progeny was determined using Southern blot analysis; herbicide resistance and SrMV resistance were assessed using bioassays. The segregation data were used to infer linkage relationships between transgenes in the parent plants. In two of the parents, all transgene insertions were linked in one position in the genome, although some recombination between insertion events did occur. In the third parent, insertion had occurred in two independent, unlinked loci. Analysis of progeny of this parent indicated that rearrangement or mutation occurred in both loci, resulting in non-parental transgene DNA fragments in some progeny. Most transgenic progeny containing the bar gene showed resistance to herbicide. SrMV inoculation indicated that a fairly high proportion of the transgenic progeny showed susceptibility. As the post-transcriptional gene silencing mechanism responsible for the virus resistance phenotype may be reset during meiosis, phenotypic screening of older plants may be a more reliable indication of virus resistance than screening young seedlings. This is the first report of transgene segregation in sugarcane, and we have demonstrated that transgenic sugarcane parents showing stable inheritance of transgenes can be effectively used in breeding programs. PMID- 12582640 TI - Construction of BAC libraries from two apomictic grasses to study the microcolinearity of their apospory-specific genomic regions. AB - We have constructed bacterial artificial chromosome (BAC) libraries from two grass species that reproduce by apospory, a form of gametophytic apomixis. The library of an apomictic polyhaploid genotype (line MS228-20, with a 2C genome size of approximately 4,500 Mbp) derived from a cross between the obligate apomict, Pennisetum squamulatum, and pearl millet ( P. glaucum) comprises 118,272 clones with an average insert size of 82 kb. The library of buffelgrass ( Cenchrus ciliaris, apomictic line B-12-9, with a 2C genome size of approximately 3,000 Mbp) contains 68,736 clones with an average insert size of 109 kb. Based on the genome sizes of these two lines and correcting for the number for false positive and organellar clones, library coverages were found to be 3.7 and 4.8 haploid genome equivalents for MS 228-20 and B12-9, respectively. Both libraries were screened by hybridization with six SCARs (sequence-characterized amplified regions), whose tight linkage in a single apospory-specific genomic region had been previously demonstrated in both species. Analysis of these BAC clones indicated that some of the SCAR markers are actually amplifying duplicated regions linked in coupling in both genomes and that restriction enzyme mapping will be necessary to sort out the duplications. PMID- 12582641 TI - Molecular characterization of wheat polyphenol oxidase (PPO). AB - It is well-established that the enzyme polyphenol oxidase (PPO) is involved in undesirable browning of noodles, chapattis, middle east flat breads and steamed breads. Methods for measuring PPO activity have been developed, and the variation of PPO activity among wheat ( Triticum aestivum L.) cultivars has been well documented. However, there is no report on the identification and characterization of a wheat PPO gene. PCR performed on wheat genomic DNA with oligonucleotide primers designed from conserved copper binding regions of other PPO genes resulted in amplification of a 444-bp DNA fragment. Sequence analysis identified the conserved amino acids of PPO genes indicating that the PCR product was part of the wheat PPO gene. Screening genomic and cDNA libraries using 444- and 760-bp DNA fragments as probes failed to identify a PPO gene based on conserved sequence, even though there were very strong hybridization signals for some isolates. Rapid amplification of cDNA ends (RACE) technique was used as an alternative to obtain the remaining DNA sequences in 5' and 3' directions based on the 444-bp partial wheat PPO gene sequence. With the use of ThermoScript Reverse Transcriptase (which functions at higher temperatures) and Advantage-GC cDNA kit, the complete DNA sequence in the 3' direction was obtained. A similar effort in the 5' direction resulted in amplification of a truncated 414-bp DNA sequence. Overall, 1,509-bp of putative wheat PPO DNA sequence was obtained. Alignment of deduced amino-acid sequences revealed similarity to the other PPO gene sequences, especially in the conserved copper binding regions. Southern-blot analysis performed with four different restriction enzymes revealed two to four DNA fragments, suggesting a limited number of PPO genes in wheat. Wheat genomic DNA restricted with HindIII and hybridized using a 760-bp wheat PPO probe revealed a clear distinction between wheat cultivars with high and low PPO activities. Northern-blot analysis indicated a transcript size of about 2.0-kb. PPO DNA fragment as well as RNA transcript was observed for the durum cultivar Renville which normally has very low PPO activity. Further study is needed to explain the relationship between PPO activity and the presence of PPO gene (s). PMID- 12582642 TI - Transpecific microsatellites for hard pines. AB - Microsatellites are difficult to recover from large plant genomes so cross specific utilisation is an important source of markers. Fifty microsatellites were tested for cross-specific amplification and polymorphism to two New World hard pine species, slash pine ( Pinus elliottii var. elliottii) and Caribbean pine ( P. caribaea var. hondurensis). Twenty-nine (58%) markers amplified in both hard pine species, and 23 of these 29 were polymorphic. Soft pine (subgenus Strobus) microsatellite markers did amplify, but none were polymorphic. Pinus elliottii var. elliottii and P. caribaea var. hondurensis showed mutational changes in the flanking regions and the repeat motif that were informative for Pinus spp. phylogenetic relationships. Most allele length variation could be attributed to variability in repeat unit number. There was no evidence for ascertainment bias. PMID- 12582643 TI - Isolation and characterization of five novel high molecular weight subunit of glutenin genes from Triticum timopheevi and Aegilops cylindrica. AB - Analysis by SDS-PAGE of total protein fractions from single seeds of Aegilops cylindrica (genomes C and D) and Triticum timopheevi (genomes A and G) showed the presence of three bands corresponding to high molecular weight subunits of glutenin (HMW subunits) in the former and two major bands and a minor band corresponding to HMW subunits in the latter. Three Ae. cylindrica and two T. timopheevi HMW subunit gene sequences, each comprising the entire coding region, were amplified by polymerase chain reaction (PCR) and their complete nucleotide sequences determined. A combination of N-terminal amino acid sequencing of the proteins identified by SDS-PAGE and alignments of the derived amino acid sequences of the proteins encoded by the PCR products identified the Ae. cylindrica HMW subunits as 1Cx, 1Cy and 1Dy, and the T. timopheevi HMW subunits as 1Gx, 1Ax and 1Ay. It was not clear whether or not a 1Gy HMW subunit was present in T. timopheevi. The PCR products from Ae. cyclindrica were derived from 1Cy and 1Dy genes and a silent 1Dx gene containing an in-frame internal stop codon, while those from T. timopheevi were derived from 1Ax and 1Ay genes. The 1Cx, 1Gx and 1Gy sequences were not amplified successfully. The proteins encoded by the five novel genes had similar structures to previously characterized HMW subunits of bread wheat ( Triticum aestivum). Differences and similarities in sequence and structure, and in the distribution of cysteine residues (relevant to the ability of HMW subunits to form high M(r) polymers) distinguished the HMW subunits of x- and y-type and of each genome rather than those of the different species. There was no evidence of a change in HMW subunit expression or structure resulting from selective breeding of bread wheat. The novel 1Ax, 1Ay, 1Cy and 1Dy HMW subunits were expressed in Escherichia coli, and the expressed proteins were shown to have very similar mobilities to the endogenous HMW subunits on SDS-PAGE. The truncated 1Dx gene from Ae. cylindrica failed to express in E. coli, and no HMW subunit-related protein of the size predicted for the truncated 1Dx subunit could be identified by immunodetection in seed extracts. PMID- 12582644 TI - Frequencies of Ty1- copia and Ty3- gypsy retroelements within the Triticeae EST databases. AB - The frequency of Ty1- copia-type and Ty3- gypsy-type retrotransposons in the International Triticeae EST Consortium (ITEC) database (61,942 sequences: 82% wheat, 10% barley, 8% rye) and the DuPont EST database (86,628 wheat sequences) was estimated using BLASTN searches. These ESTs were obtained from 94 cDNA libraries from different tissues (leaves, roots, spikes, flowers and seeds) and different growing conditions, excluding subtracted and normalized cDNA libraries. Triticeae EST databases were screened using four different Ty1- -copia-type, 12 reverse transcriptase sequences, and three Ty3- gypsy-type Triticeae retrotransposon sequences. Using a selection threshold of BLASTN scores higher than 100 or E values smaller than e(-20), 0.145% of the ESTs were found to be significantly similar to at least one of the retrotransposons used in the search (0.064% Ty1- copia, 0.081% Ty3- gypsy). This percentage increased to 0.176% when the BLASTN threshold was changed to E2.0) for partial resistance was detected on chromosome 11. This QTL explained 45.6% of the phenotypic variation. The segregation ratio of the F3 lines was 3:1 for partial resistance to susceptibility. These results suggest that the partial resistance in Chubu 32 is controlled by a major gene. PMID- 12582658 TI - The relationship between homozygous and hemizygous transgene expression levels over generations in populations of transgenic rice plants. AB - Segregating T(1), T(2) and T(3) transgenic rice populations, derived from independent particle-bombardment-mediated transformation events were examined in order to assess the effect of gene dosage on transgene expression levels and stability. The expression level of the unselected beta-glucuronidase ( gusA) reporter gene was quantified in plants from these populations. The gusA gene dosage was determined by segregation analysis of progeny seedlings at the structural level (by PCR) and at the expression level. For some transformation events a gene dosage effect on transgene expression was observed, leading to higher transgene expression levels in homozygous progeny than in hemizygous progeny or primary transgenic plants. However, in many other transformation events, the homozygous state appears to be disadvantageous, being associated with lower transgene expression levels, gene silencing or counter-selection of homozygous plants across generations. Change of gene dosage is probably one of the key factors influencing transgene expression levels and stability in transgenic rice. This is particularly important when considering molecular genetic studies and crop improvement programmes. The possible influence of matrix attachment regions (MARs) in increasing the likelihood of an additive effect on transgene expression level is discussed. PMID- 12582659 TI - Molecular diversity of male sterility inducing and male-fertile cytoplasms in the genus Helianthus. AB - The organisation of mtDNA was investigated for 28 sources of cytoplasmic male sterility (CMS) and a fertile line (normal cytoplasm) of Helianthus annuus by Southern hybridisation. In addition to nine known mitochondrial genes ( atp6, atp9, cob, coxI, coxII, coxIII, 18S, 5S and nd5) three probes for the open reading frames in the rearranged area of PET1, orfH522, orfH708 and orfH873, were used. Genetic similarities of the investigat-ed cytoplasms varied between 0.3 and 1. Cluster analyses using the UPGMA method allowed the distinction of ten mitochondrial (mt) types between the 29 investigated cytoplasms. Most mitochondrial types comprise two or more CMS sources, which could not be further separated, like the PET1-like CMS sources (with the exception of ANO1 and PRR1), or ANN1/ANN2/ANN3, ANN4/ ANN5, ARG3/RIG1, BOL1/EXI1/PEF1/PEP1 and GIG1/ PET2. ANL1, ANL2 and the fertile cytoplasms are also regarded as one mitochondrial type. Unique banding patterns were only observed for ANT1 ( atp6), MAX1 ( atp6, orfH522 and orfH708) and PRR1 ( coxII). However, four of the mitochondrial types showed unique hybridisation signals: ANN4/ANN5 had characteristic bands for atp6 and orfH708, PEF1/PEP1/EXI1/BOL1 for atp6and coxII, and PET2/GIG1 for atp9. The PET1-like cytoplasms all shared the same patterns for orfH522, orfH708and cob (except ANO1). It could be demonstrated that CMS sources, like, e.g., PET2 and PEF1, are different from PET1 in mtDNA organisation and the CMS mechanism. Therefore, these CMS sources represent interesting candidates for the development of new hybrid breeding systems based on new CMS mechanisms. PMID- 12582660 TI - Genetic analysis of Vrn-B1 for vernalization requirement by using linked dCAPS markers in bread wheat ( Triticum aestivum L.). AB - To identify a molecular marker closely linked to Vrn-B1, the Vrn-1 ortholog on chromosome 5B, sequence polymorphism at four orthologous RFLP loci closely linked to the Vrn-1 gene family was analyzed by using near-isogenic lines of "Triple Dirk." At Xwg644, a RFLP locus, three types of nucleotide sequence differing by the number of (TG) repeats, two or three times, and base changes were detected. A (TG)(3)-type sequence proved to be specific to chromosome 5B by nulli-tetrasomic analysis, and substitution of single nucleotide (C/T) was detected between TD(B) carrying the former Vrn2 allele and TD(C) carrying the vrn2 allele. A mismatch primer was designed for dCAPS analysis of this single nucleotide polymorphism (SNP). Polymorphism was successfully detected between two NILs, through nested PCR by using a (TG)(3)-specific primer (1st) and a dCAPS primer (2nd) followed by a NsiI digest. The analysis of a BF(2) population [(TD(B)//TD(C)] revealed the close linkage (1.7 cM) between WG644-5B and Vrn2. It was therefore concluded that the former Vrn2 locus is located on chromosome 5B and equivalent to Vrn-B1. PMID- 12582661 TI - Development and mapping of AFLP markers linked to the sorghum fertility restorer gene rf4. AB - The restoration of male fertility in the sorghum IS1112 C (A3) male-sterile cytoplasm is through a two-gene gametophytic system involving complementary action of the restoring alleles Rf3 and Rf4. To develop markers suitable for mapping rf4, AFLP technology was applied to bulks of sterile and fertile individuals from a segregating BC(3)F(1) population. Three AFLP markers linked to rf4were identified and subsequently converted to STS/CAPS markers, two of which are co-dominant. Based on a population of 378 BC(1)F(1) individuals, two STS/CAPS markers, LW7 and LW8, mapped to within 5.31 and 3.18 cM, respectively, of rf4, while an STS marker, LW9, was positioned 0.79 cM on the flanking side of rf4. Markers LW8 and LW9 were used to screen sorghum BAC libraries to identify the genomic region encoding rf4. A series of BAC clones shown to represent a genomic region of linkage group E were identified by the rf4-linked markers. A contig of BAC clones flanking the LW9 marker represent seed clones on linkage group E, from which fine mapping of the rf4 locus and chromosome walking can be initiated. PMID- 12582662 TI - QTLs involved in the restriction of cucumber mosaic virus (CMV) long-distance movement in pepper. AB - Partial restriction of cucumber mosaic virus (CMV) long-distance movement originating from the Capsicum annuum inbred line 'Vania' was assessed in a doubled-haploid progeny using two screening methods: the first allowed one to assess the resistance of adult plants decapitated above the fourth leaf and inoculated on the third leaf using a common CMV strain, and the second allowed one to assess CMV resistance to long-distance movement on seedlings inoculated using an atypical CMV strain. For both resistance tests, the behavior of the F(1) hybrid between 'Vania' and the susceptible line 'H3' indicated that partial resistance is inherited as a dominant trait. Phenotypic data from the two screening methods were correlated but the one performed on seedlings was much more severe. A subset of 184 molecular markers well-distributed over the pepper genome was selected for QTL mapping using the composite interval mapping (CIM) method. A total of seven genomic regions, including one major effect and several minor effect QTLs, were shown to be associated with partial restriction of CMV long-distance movement. These results are compared with those already obtained in pepper and also in other solanaceous crops, potato and tomato. PMID- 12582663 TI - Molecular analysis of a major locus for resistance to downy mildew in sunflower with specific PCR-based markers. AB - Resistance of sunflower to the obligate parasite Plasmopara halstedii is conferred by specific dominant genes, denoted Pl. The Pl6 locus confers resistance to all races of P. halstedii except one, and must contain at least 11 tightly linked genes each giving resistance to different downy mildew races. Specific primers were designed and used to amplify 13 markers covering a genetic distance of about 3 cM centred on the Pl6 locus. Cloning and sequence analysis of these 13 markers indicate that Pl6 contains conserved genes belonging to the TIR NBS-LRR class of plant resistance genes. PMID- 12582664 TI - Identification of quantitative trait loci controlling developmental characteristics of Brassica oleracea L. AB - A segregating population of F(1)-derived doubled haploid (DH) lines of Brassica oleracea was used to detect and locate QTLs controlling 27 morphological and developmental traits, including leaf, flowering, axillary bud and stem characters. The population resulted from a cross between two very different B. oleracea crop types, an annual cauliflower and a biennial Brussels sprout. A principal component analysis (PCA), based on line means, allowed all the traits to be grouped into distinct categories according to the first five Principal Components. These were: leaf traits (PC1), flowering traits (PC2), axillary bud traits (PC3 and 5) and stem traits (PC4). Between zero and four putative QTL were located per trait, which individually explained between 6% and 43% of the additive genetic variation, using the multiple-marker regression approach to QTL mapping. For lamina width, bare petiole length and stem length two QTL with opposite effects were detected on the same linkage groups. Intra- and inter specific comparative mapping using RFLP markers identified a QTL on linkage group O8 accounting for variation in vernalisation, which is probably synonymous with a QTL detected on linkage group N19 of Brassica napus. In addition, a QTL for petiole length detected on O3 of this study appeared to be homologous to a QTL detected on another B. oleracea genetic map (Camargo et al. 1995). PMID- 12582665 TI - Identification of resistance gene analogs linked to a powdery mildew resistance locus in grapevine. AB - Oligonucleotide primers, designed to conserved regions of nucleotide binding site (NBS) motifs within previously cloned pathogen resistance genes, were used to amplify resistance gene analogs (RGAs) from grapevine. Twenty eight unique grapevine RGA sequences were identified and subdivided into 22 groups on the basis of nucleic acid sequence-identity of approximately 70% or greater. Representatives from each group were used in a bulked segregant analysis strategy to screen for restriction fragment length polymorphisms linked to the powdery mildew resistance locus, Run1, introgressed into Vitis vinifera L. from the wild grape species Muscadinia rotundifolia. Three RGA markers were found to be tightly linked to the Run1 locus. Of these markers, two (GLP1-12 and MHD145) cosegregated with the resistance phenotype in 167 progeny tested, whereas the third marker (MHD98) was mapped to a position 2.4 cM from the Run1 locus. The results demonstrate the usefulness of RGA sequences, when used in combination with bulked segregant analysis, to rapidly generate markers tightly linked to resistance loci in crop species. PMID- 12582666 TI - Identification of quantitative trait loci and epistatic interactions for plant height and heading date in rice. AB - Appropriate heading date and plant height are prerequisites for attaining the desired yield level in rice breeding programs. In this study, we analyzed the genetic bases of heading date and plant height at both single- locus and two locus levels, using a population of 240 F(2:3) families derived from a cross between two elite rice lines. Measurements for the traits were obtained over 2 years in replicated field trials. A linkage map was constructed with 151 polymorphic marker loci, based on which interval mapping was performed using Mapmaker/QTL. The analyses detected six QTLs for plant height and six QTLs for heading date; collectively the QTLs for heading date accounted for a much greater amount of phenotypic variation than did the QTLs for plant height. Two-way analyses of variance, with all possible two-locus combinations, detected large numbers (from 101 to 257) of significant digenic interactions in the 2 years for both traits involving markers distributed in the entire genome; 22 and 39 were simultaneously detected in both years for plant height and heading date, respectively. Each of the interactions individually accounted for only a very small portion of the phenotypic variation. The majority of the significant interactions involved marker loci that did not detect significant effects by single-locus analyses, and many of the QTLs detected by single-locus analyses were involved in epistatic interactions. The results clearly demonstrated the importance of epistatic interactions in the genetic bases of heading date and plant height. PMID- 12582667 TI - Molecular mapping, phenotypic expression and geographical distribution of genes determining anthocyanin pigmentation of coleoptiles in wheat ( Triticum aestivumL.). AB - Three major gene loci determining the anthocyanin pigmentation of coleoptiles were mapped on the short arms of chromosomes 7A, 7B and 7D, respectively. All three genes map about 15 to 20 cM distal from the centromere and, therefore, it may be concluded that they are members of a homoeologous series and should be designated Rc-A1, Rc-B1 and Rc-D1, respectively. Further homoeologous loci exist in Triticum durum, Triticum tauschii, and most probably in Secale cereale and Hordeum vulgare. By analyzing a syntheticxcultivated wheat cross (ITMI mapping population) under different environmental conditions it was shown that the expression of the genes determining anthocyanin pigmentation of the coleoptiles varies. One additional locus was detected on chromosome 4BL. Beside the mapping data, results of a screening for red coleoptile color genes in 468 mainly European wheat varieties are presented. PMID- 12582668 TI - Pollen competition and seed-siring success in Picea abies. AB - The aim of the present work was to study pollen-tube competition in Picea abies. Controlled crossings were performed with pollen mixtures including pairs of pollen lots with fast and slowly elongating pollen-tubes. Paternity analysis using isozyme markers was performed on the progenies in order to study whether the in vitro pollen-germination vigour corresponds to the proportion of seeds sired by the pollen donor. Paternal success was found to be unequal, 15 out of 23 crossings producing progeny that differed significantly from the hypothetical ratio of 1:1. The paternal contribution in the majority of the crossings was as expected: the pollen parent with more-vigorous in vitro germination sired more seeds than the less-vigorous pollen. In the case of two pollen mixtures, however, the seed-siring success summed over the maternal trees was the opposite to the expected value. Despite these aberrations, the results support the hypothesis that pollen-tube competition is one of the factors contributing to male fitness in P. abies.However, when all the other factors affecting pollination and seed set under natural conditions are taken into account, it is clear that the seed siring success of a particular paternal genotype cannot be predicted reliably by measuring only the in vitro pollen vigour. PMID- 12582669 TI - Phylogenetic analysis in the genus Cicer and cultivated chickpea using RAPD and ISSR markers. AB - Seventy five accessions belonging to 14 species of the genus Cicer were analysed with PCR-based molecular markers to determine their phylogenetic relationships. Eight of the species were annuals and included the Section Monocicer which contains cultivated chickpea ( Cicer arietinum L.). The remaining six species were perennials (five from Section Polycicer and one from Section Acanthocicer). More than one accession per species was analysed in most of the wild species; within C. arietinum, 26 accessions including Kabuli and Desi types, were studied. RAPD analyses using 12 primers gave 234 polymorphic fragments. Variability within species was detected. A dendrogram based on the Jaccard similarity index showed that the distribution pattern of variability between species was related to both growth habit and geographical origin. An accession of Cicer reticulatum was closer to accessions of Cicer echinospermum than to the four remaining of C. reticulatum, suggesting the possibility of gene flow between species. Cluster analysis for cultivated chickpea differentiated Kabuli and Desi types but we did not detect a clear relationship between groups and the geographical origin of the accessions. PMID- 12582670 TI - Occurrence of partial hybrids in wide crosses between sunflower ( Helianthus annuus) and perennial species H. mollisand H. orgyalis. AB - Hybridisation between the annual diploid sunflower ( Helianthus annuus)and the perennial diploid species Helianthus mollis and Helianthus orgyalis was obtained by means of a normal crossing procedure or embryo rescue. Hybridisation success was low. All plants examined cytologically appeared to be diploid. However, the phenotypes of these diploids were not intermediate between the parents and, despite great variation, they resembled the female parent-type predominantly. Thirty five percent of plants issued from sunflower pollinated with perennial Helianthus had a phenotype resembling the female sunflower parent. On average, only 5% of the minimum number of expected RAPD and RFLP bands from male parents were recovered in plants produced from mature seeds after pollination of sunflower by H. mollis. More hybrids were found among plants obtained from embryo rescue, with an average of 25% of the male parent bands recovered per plant. Analysis of individual plants indicated the occurrence of various levels of hybridisation. There was a significant positive correlation between the number of phenotype traits related to hybrid status and the number of bands derived from the male parent. A single hybrid plant might possibly represent the product of a 'normal' hybridisation event. The mechanisms behind these unusual events and the consequences for the breeder are discussed. PMID- 12582671 TI - Introgression into the allotetraploid coffee ( Coffea arabica L.): segregation and recombination of the C. canephora genome in the tetraploid interspecific hybrid ( C. arabicax C. canephora). AB - Transfer of desired characters from the diploid relative species such as Coffea canephora into the cultivated allotetraploid coffee species ( Coffea arabica L.) is essential to the continued improvement of varieties. Behaviour of the C. canephora genome and its interaction with the C. arabica genome were investigated in tetraploid interspecific hybrids ( C. arabicax C. canephora 4 x) resulting from a cross between an accession of C. arabica and a tetraploid plant of C. canephora obtained following colchicine treatment. Segregation and co-segregation of restriction fragment length polymorphism (RFLP) and microsatellite loci markers were studied in two BC(1) populations. These two populations of 28 and 45 individuals, respectively, resulted from the backcross of two tetraploid F(1)plants to C. arabica. The presence in BC(1) plants of specific C. canephora markers was scored for 24 loci (11 RFLP and 13 microsatellites) distributed on at least 7 of the 11 linkage groups identified in C. canephora. At almost all loci analysed, the segregation of C. canephora alleles transmitted by the ( C. arabicax C. canephora 4 x) hybrids conformed to the expected ratio assuming random chromosome segregation and the absence of selection. The recombination fractions of C. canephorachromosome segments were estimated for seven marker intervals, and compared with the recombination fractions previously observed in C. canephora for the equivalent marker intervals. The recombination frequencies estimated in both plant materials were rather similar, suggesting that recombination in the ( C. arabicax C. canephora 4 x) hybrid is not significantly restricted by the genetic differentiation between chromosomes belonging to the different genomes. The hybrid ( C. arabicax C. canephora 4 x) therefore appeared particularly favourable to intergenomic recombination events and gene introgressions. PMID- 12582672 TI - A new gene, Ny (tbr), for hypersensitivity to Potato virus Y from Solanum tuberosumMaps to Chromosome IV. AB - A diploid backcross population derived from a cross between Solanum tuberosum and Solanum berthaultiisegregated for monogenic dominant hypersensitivity to Potato virus Y(PVY). We propose the symbol Ny (tbr) for this locus because plants carrying this gene develop necrosis after inoculation with PVY and the allele originated in S. tuberosum. The gene mapped to chromosome IV between TG316 and TG208 at LOD=2.72. This location does not correspond to any other mapped resistance genes in potato. PMID- 12582673 TI - Hierarchical and factorial mating designs for quantitative genetic analysis in tetrasomic potato. AB - Plant breeders need to quantify additive and non-additive components of genetic variance in order to determine appropriate selection methods to improve quantitative characteristics. Hierarchical and factorial mating designs (also known as North Carolina mating designs I and II, respectively) allow one to determine these variance components. The relative advantages of these two designs in the quantitative genetics of tuber yield in tetrasomic potato were investigated. Likewise, the number of female parents to include in design I was also investigated. Data were collected from two independent experiments at two contrasting Peruvian locations: La Molina in the dry coast and San Ramon in the humid mid-altitude. In the first experiment, although design I gave a negative digenic variance (sigma(2)(D)), this design provided almost the same estimate of narrow-sense heritability (h(2)) for tuber yield as that obtained in design II (0.291 and 0.260, respectively). Therefore, design I appears to be appropriate for quantitative genetics research in tetrasomic potato, a crop in which some clones are male sterile. The easy handling of crosses (distinct random females included in the crossing scheme) is another advantage of design I relative to design II. In the second experiment, 12 males were crossed with either two or four females following a design-I mating scheme. The additive genetic variance (sigma(2)(A)) was zero (or negative) when two females per male were included but was positive with four females. These results suggest that two females per male may not be enough for design I in tetrasomic potato. Four females per male are preferable to determine sigma(2)(A) in design I for this tetrasomic crop. PMID- 12582674 TI - Identification of new low-molecular-weight glutenin subunit genes in wheat. AB - To clarify the composition of low-molecular-weight glutenin subunits (LMW-GSs) in a soft wheat cultivar, we cloned and characterized LMW-GS genes from a cDNA library and genomic DNA in Norin 61. Based on alignment of the conserved N- and C terminal domains of the deduced amino-acid sequences, these genes are classified into 12 groups. One of these groups (group 5), the corresponding gene of which has not been reported previously, contains two additional hydrophobic amino-acid clusters interrupting the N-terminal repetitive domain. Other groups (groups 11 and 12), which were not identified in other cultivars as a protein product, showed all eight cysteines in the C-terminal conserved domain. With specific primer sets for these groups it was revealed that Glu-D3 and Glu-A3 encoded the former and the latter, respectively. Both groups of genes were expressed in immature seeds. The presence of these groups of LMW-GSs may affect the dough strength of soft wheat. PMID- 12582675 TI - Crop-weed interactions in the Beta vulgaris complex at a local scale: allelic diversity and gene flow within sugar beet fields. AB - Crop-wild hybrids and weed beets are the main source of agronomic concern for sugar beet production all over Europe. In order to understand the dynamics of crop-wild interactions and the evolution of weediness in Beta vulgaris, we investigated genetic features of bolting individuals occurring at a local scale, i.e. within two sugar beet fields of the French northern area of sugar beet production. By analysing ploidy level, mitochondrial DNA and microsatellite polymorphism, the genetic diversity and the genetic relationships among three different classes of individuals (variety, in-row and out-row weed-beets) from a given field were examined. Such genetic analyses provide a unique opportunity to obtain evidence for the weeds origin and the evolutionary hypotheses previously stated. All the individuals shared in common the Svulg mitochondrial haplotype, and thus a common maternal origin. Conversely, the large genetic diversity at microsatellite loci highlighted the large diversity of the pollinator plants (cultivated and wild plants) during the-seed production process, as well as during the further evolution of weed beets in the sugar production area. PMID- 12582676 TI - A PCR-marker for the CMS(1) inducing cytoplasm in chives derived from recombination events affecting the mitochondrial gene atp9. AB - The complete coding and 3;-flanking region of the mitochondrial gene atp9 of chives ( Allium schoenoprasum L.) was determined in order to develop primers that allow the identification of atp9-related sequences in subsequent PCR amplifications. One of these sequences is of a chimerical nature, consisting of atp9-homologous regions on its end, interrupted by an insertion that is composed of one atp6-homologous part and one part of unknown origin. This PCR-fragment is 762 bp in size and exclusively amplified in the sterility inducing cytoplasm of CMS(1). Thus it can be used as a PCR-marker in order to distinguish this cytoplasm type from the remaining cytoplasm types of chives. The chimerical marker sequence forms a putative open reading frame ( orfA501), from which CMS(1) might originate. PMID- 12582677 TI - In vivo haploid induction in maize. AB - Two haploid-inducing lines, MHI and M741H, were used for the production of maternal haploids. Haploids were obtained from all maternal genotypes involved in the experiment, including dent, flint and flintxdent maize. The maternal genotype had a significant influence on the frequency of haploids obtained. The frequency ranged from 2.7% to 8.0%. For chromosome-doubling seedlings were treated with colchicine solution, and 49.4% of the haploid plants produced fertile pollen, 39.0% could be selfed and 27.3% produced seeds after selfing. Synthetic populations, improved by haploid sib recurrent selection, were tested in a field trial. The results show that the utilization of maternal haploid plants has great potential for maize breeding and maize genetics. PMID- 12582678 TI - Efficient microprojectile bombardment-mediated transformation of rice using gene cassettes. AB - This study was aimed at determining whether gene cassettes (promoter-coding sequence-terminator) can be efficiently used in microprojectile acceleration mediated co-transformation of rice in the place of whole plasmids, and to what extent their use influences the integration and expression of the co-transferred gene of interest. Two non-linked marker genes ( yfp and hph) were co-introduced by microprojectile bombardment into cells of embryogenic calli in three separate experiments. Three different DNA structures were compared for their ability to transiently and stably transform rice cells: supercoiled or linearized whole plasmid DNA, gene cassette DNA and single-stranded gene cassette DNA coated with Escherichia coli single-stranded binding (SSB) proteins. Our results demonstrate that microprojectile bombardment-mediated transformation of rice using gene cassettes is possible without significantly reducing transformation efficiency in comparison to the use of whole-plasmid DNA. Furthermore, no obvious difference in transgene integration pattern and inheritance was observed among plants transformed with gene cassettes compared to those transformed with the whole plasmid, except that concatemerization of molecules prior to integration was rarely observed in gene cassette transformants. PMID- 12582679 TI - PCR detection of Hordeum bulbosum introgressions in an H. vulgare background using a retrotransposon-like sequence. AB - Retrotransposon-like sequences are ideal tools for initial screening assays to distinguish between closely related species because of their ubiquitous presence, high copy number, chromosome coverage and rapid sequence evolution. A retrotransposon-like sequence, pSc119.1, cloned from Secale cereale (rye) has been used to obtain PCR primers that are capable of detecting small introgressions of Hordeum bulbosum (bulbous barley grass) chromatin in a Hordeum vulgare (cultivated barley) background. Combining this PCR-based assay with a crude but effective high-throughput DNA extraction has enabled the rapid identification of plants possessing H. bulbosumintrogressions from large numbers of progeny from H. vulgarex H. bulbosumcrosses. These plants are then further characterized by more-refined cytological, molecular and pathological techniques to locate and map the introgressed chromatin and to evaluate their disease resistance. PMID- 12582680 TI - Inheritance and expression of the cry1Ab gene in Bt ( Bacillus thuringiensis) transgenic rice. AB - The inheritance and expression patterns of the cry1Ab gene were studied in the progenies derived from different Bt ( Bacillus thuringiensis) transgenic japonica rice lines under field conditions. Both Mendelian and distorted segregation ratios were observed in some selfed and crossed F(2) populations. Crosses between japonica intra-subspecies had no significant effect on the segregation ratios of the cry1Ab gene, but crossing between japonica and indicainter-subspecies led to distorted segregation of the cry1Ab gene in the F(2)population. Field-release experiments indicated that the cry1Ab gene was stably transmitted in an intact manner via successive sexual generations, and the concentration of the Cry1Ab protein was kept quantitatively stable up to the R(6)generation. The cry1Ab gene, driven by the maize ubiquitinpromoter, displayed certain kinds of spatial and temporal expression patterns under field conditions. The content of the Cry1Ab protein varied in different tissues of the main stems, the primary tillers and the secondary tillers. Higher levels of the Cry1Ab protein were found in the stems, leaves and leaf sheaths than in the roots, while the lowest level was detected in grains at the maturation stage. The content of the Cry1Ab protein in the leaves peaked at the booting stage and was lowest at the heading stage. Furthermore, the Cry1Ab content of cry1Ab expression in different tissues of transgenic rice varied individually with temperature. PMID- 12582681 TI - Utility of barley and wheat simple sequence repeat (SSR) markers for genetic analysis of Hordeum chilense and tritordeum. AB - A selection of 36 wheat and 35 barley simple sequence repeat markers (SSRs) were studied for their utility in Hordeum chilense. Nineteen wheat and nineteen barley primer pairs amplified consistent H. chilense products. Nine wheat and two barley SSRs were polymorphic in a H. chilense mapping population, producing codominant markers that mapped to the expected homoeologous linkage groups in all but one case. Thirteen wheat and 10 barley primer pairs were suitable for studying the introgression of H. chilense into wheat because they amplified H. chilense products of distinct size. Analysis of wheat/ H. chilense addition lines showed that the H. chilense products derived from the expected homoeologous linkage groups. The results showed that wheat and barley SSRs provide a valuable resource for the genetic characterization of H. chilense, tritordeums and derived introgression lines. PMID- 12582682 TI - Genetic analysis and FISH mapping of the Colourless non-ripening locus of tomato. AB - Cnr ( Colourless non-ripening) is a dominant pleiotropic ripening mutation of tomato ( Lycopersicon esculentum) which has previously been mapped to the proximal region of tomato chromosome 2. We describe the fine mapping of the Cnr locus using both linkage analysis and fluorescence in situ hybridisation (FISH). Restriction fragment length polymorphism (RFLP)-, amplified restriction fragment polymorphism (AFLP)-, and cleaved amplified polymorphic sequence (CAPS)-based markers, linked to the Cnrlocus were mapped onto the long arm of chromosome 2. Detailed linkage analysis indicated that the Cnr locus was likely to lie further away from the top of the long arm than previously thought. This was confirmed by FISH, which was applied to tomato pachytene chromosomes in order to gain an insight into the organisation of hetero- and euchromatin and its relationship to the physical and genetic distances in the Cnr region. Three molecular markers linked to Cnr were unambiguously located by FISH to the long arm of chromosome 2 using individual BAC probes containing these single-copy sequences. The physical order of the markers coincided with that established by genetic analysis. The two AFLP markers most-closely linked to the Cnr locus were located in the euchromatic region 2.7-cM apart. The physical distance between these markers was measured on the pachytene spreads and estimated to be approximately 900 kb, suggesting a bp:cM relationship in this region of chromosome 2 of about 330 kb/cM. This is less than half the average value of 750 kb/cM for the tomato genome. The relationship between genetic and physical distances on chromosome 2 is discussed. PMID- 12582683 TI - Production of a new wheat line possessing the 1BL.1RS wheat-rye translocation derived from Korean rye cultivar Paldanghomil. AB - The 1BL.1RS translocations between wheat ( Triticum aestivum L.) and rye ( Secale cereale L.) are widely used in bread wheat breeding programs, but all modern wheat cultivars with the 1BL.1RS have shown genetic vulnerability due to one rye source - a German cultivar, Petkus. We have developed, a new 1BL.1RS wheat-rye translocation line from the backcross of the F(1) hybrid of wheat cv. Olmil and rye cv. Paldanghomil, both cultivars from Korea. The GISH technique was applied to identify the presence of rye chromatin in 467 BC(1)F(6) lines selected from 77 BC(1)F(5) lines. Only one line, Yw62-11, showed wheat-rye translocated chromosomes, with a somatic chromosome number of 2n=42. C-banding patterns revealed that the translocated chromosome was 1BL.1RS, showing prominent bands in the terminal and sub-terminal regions of the short arm as well as in the centromeric region and terminal region of the long arm. This new 1BL.1RS translocation line formed 21 bivalents like common wheat at meiotic metaphase I, thereby showing complete homology. PMID- 12582684 TI - A genome-specific repetitive DNA sequence from Oryza eichingeri: characterization, localization, and introgression to O. sativa. AB - In the course of transferring the brown planthopper resistance from a diploid, CC genome wild rice species, Oryza eichingeri (IRGC acc. 105159 and 105163), to the cultivated rice variety 02428, we have isolated many alien addition and introgression lines. The O. eichingeri chromatin in some of these lines has previously been identified using genomic in situ hybridization and molecular marker analysis. Here we cloned a tandemly repetitive DNA sequence from O. eichingeri IRGC acc105163, and detected it in 25 introgression lines. This repetitive DNA sequence showed high specificity to the rice CC genome, but was absent from all the four tetraploid species with BBCC or CCDD genomes. The monomer in this repetitive DNA sequence is 325-366-bp long, with a copy number of about 5,000 per 1 C of the O. eichingerigenome, showing 88% homology to a repetitive DNA sequence isolated from Oryza officinalis(2n=2 x=24, CC). Fluorescent in situ hybridization revealed 11 signals distributed over eight O. eichingeri chromosomes, mostly in terminal or subterminal regions. PMID- 12582685 TI - Physical location of a HSP70 gene homologue on the centromere of chromosome 1B of wheat ( Triticum aestivum L.). AB - Cereal centromeres consist of a complex organization of repetitive DNA sequences. Several repetitive DNA sequences are common amongst members of the Triticeae family, and others are unique to particular species. The organization of these repetitive elements and the abundance of other types of DNA sequences in cereal centromeres are largely unknown. In this study, we have used wheat-rye translocation lines to physically map 1BL.1RS centromeric breakpoints and molecular probes to obtain further information on the nature of other types of centromeric DNA sequences. Our results, using the rye-specific centromeric sequence, pAWRC.1, indicate that 1BL.1RS contains a small portion of the centromere from 1R of rye. Further studies used molecular markers to identify centromeric segments on wheat group-1 chromosomes. Selected RFLP markers, clustered around the centromere of wheat homoeologous group-1S chromosomes, were chosen as probes during Southern hybridization. One marker, PSR161, identified a small 1BS segment in all 1BL.1RS lines. This segment maps proximal to pAWRC.1 in 1BL.1RS and on the centromere of 1B. Sequence analysis of PSR161 showed high homology to HSP70 genes and Northern hybridization showed that this gene is constitutively expressed in leaf tissue and induced by heat shock and light stimuli. The significance of this work with respect to centromere organization and the possible significance of this HSP70 gene homologue are discussed. PMID- 12582686 TI - Chimerism in grapevines: implications for cultivar identity, ancestry and genetic improvement. AB - In the course of DNA profiling of grapevine cultivars using microsatellite loci we have occasionally observed more than two alleles at a locus in some individuals and have identified periclinal chimerism as the source of such anomalies. This phenomenon in long-lived clonally propagated crops, such as grapevine, which contains historically ancient cultivars, may have a role in clonal differences and affect cultivar identification and pedigree analysis. Here we show that when the two cell layers of a periclinal chimera, Pinot Meunier, are separated by passage through somatic embryogenesis the regenerated plants not only have distinct DNA profiles which are different from those of the parent plant but also have novel phenotypes. Recovery of these phenotypes indicates that additional genetic differences can exist between the two cell layers and that the Pinot Meunier phenotype is due to the interaction of genetically distinct cell layers. It appears that grapevine chimerism can not only modify phenotype but can also impact on grapevine improvement as both genetic transformation and conventional breeding strategies separate mutations in the L1 and L2 cell layers. PMID- 12582687 TI - RFLP genetic linkage maps from four F(2.3) populations and a joinmap of Gossypium hirsutum L. AB - An RFLP genetic linkage joinmap was constructed from four different mapping populations of cotton ( Gossypium hirsutum L.). Genetic maps from two of the four populations have been previously reported. The third genetic map was constructed from 199 bulk-sampled plots of an F(2.3) (HQ95-6x'MD51ne') population. The map comprises 83 loci mapped to 24 linkage groups with an average distance between markers of 10.0 centiMorgan (cM), covering 830.1 cM or approximately 18% of the genome. The fourth genetic map was developed from 155 bulk-sampled plots of an F(2.3) (119- 5 sub-okrax'MD51ne') population. This map comprises 56 loci mapped to 16 linkage groups with an average distance between markers of 9.3 cM, covering 520.4 cM or approximately 11% of the cotton genome. A core of 104 cDNA probes was shared between populations, yielding 111 RFLP loci. The constructed genetic linkage joinmap from the above four populations comprises 284 loci mapped to 47 linkage groups with the average distance between markers of 5.3 cM, covering 1,502.6 cM or approximately 31% of the total recombinational length of the cotton genome. The linkage groups contained from 2 to 54 loci each and ranged in distance from 1.0 to 142.6 cM. The joinmap provided further knowledge of competitive chromosome arrangement, parental relationships, gene order, and increased the potential to map genes for the improvement of the cotton crop. This is the first genetic linkage joinmap assembled in G. hirsutum with a core of RFLP markers assayed on different genetic backgrounds of cotton populations (Acala, Delta, and Texas plain). Research is ongoing for the identification of quantitative trait loci for agronomic, physiological and fiber quality traits on these maps, and the identification of RFLP loci lineage for G. hirsutum from its diploid progenitors (the A and D genomes). PMID- 12582688 TI - The sugar beet mitochondrial nad4 gene: an intron loss and its phylogenetic implication in the Caryophyllales. AB - The sugar beet mitochondrial gene for subunit IV of NADH dehydrogenase ( nad4) has been characterized. Unlike the corresponding genes in wheat and turnip, sugar beet nad4 lacks the second intron ( nad4-i2). Northern-blot analysis demonstrates transcription of the gene. A total of 19 RNA editing sites were identified in the sugar beet nad4 transcripts; interestingly, there is no editing in the region which flanks the lost intron. This observation is in favour of intron loss via homologous recombination of an edited RNA intermediate. We also found that the nad4-i2 intron is absent from the mitochondrial genomes of all examined members of the Caryophyllales, but present in the closely related orders, Polygonales and Plumbaginales, which suggests that the intron was lost in the common ancestor of the Caryophyllales. PMID- 12582689 TI - Identification of QTLs influencing wood property traits in loblolly pine ( Pinus taeda L.). II. Chemical wood properties. AB - Chemical wood property traits were analyzed for the presence of quantitative trait loci (QTLs) in a three-generation outbred pedigree of loblolly pine ( Pinus taeda L.). These traits were assayed using pyrolysis molecular beam mass spectrometry and include mass spectrum peak intensities associated with carbohydrates, alpha-cellulose and hemicellulose sugars, and lignin. Models for projection to latent structures (PLS) were used to also estimate the chemical composition of cell walls (i.e., alpha-cellulose, galactan and lignin) from mass spectrum data using multivariate regression. Both earlywood and latewood fractions from the fifth annual ring were analyzed for each trait. An interval mapping approach designed for an outbred pedigree was used to estimate the number of QTLs, the magnitude of QTL effects, and their genomic position. Eight unique QTLs influencing cell wall chemistry were detected from multiple peak intensities and/or PLS estimates using the one- and two-QTL models. Significant differences in chemical contents were observed among the populations from North Carolina vs Oklahoma, and results from QTLxenvironment analyses suggest that QTLs interact with environmental location. QTLs should be verified in larger experiments and in different genetic and environmental backgrounds. QTL mapping will help towards eventually identifying genes having a major effect on chemical wood properties. PMID- 12582690 TI - Microsatellite markers isolated in olive ( Olea europaea L.) are suitable for individual fingerprinting and reveal polymorphism within ancient cultivars. AB - We have isolated and sequenced 52 microsatellites or simple sequence repeats (SSRs) from nearly 60 positive clones obtained from two 'Frantoio' olive genomic libraries enriched in (AC/GT) and (AG/CT) repeats, respectively. The repeat containing fragments obtained from genomic DNA restricted with Tsp509I were separated using a biotinylated probe bound to streptavidin-coated paramagnetic beads. Fragments were then cloned into lambda ZAPII vector and sequenced. Thirty of the 36 primer pairs which gave correct re-amplification in the source genome were used to assay the polymorphism of 12 olive cultivars, namely four well-known cultivars ('Coratina', 'Frantoio', 'Leccino', 'Pendolino') and eight ancient cultivars grown locally near Lake Garda ('Casaliva', 'Favarol', 'Fort', 'Grignan', 'Less', 'Raza', 'Rossanel', 'Trep'). The local cultivars were each re- presented by two to four long-lived individuals. The analysis was carried out using (33)P-labelled primers and 6% polyacrylamide sequencing gels. All except two microsatellites showed polymorphism, the number of alleles varying from 1 to 5. The average genetic diversity ( H) was 0.55. The power of discrimination ( PD) was 0.60. All cultivars, including the local ones, were easily separated from each other. Variations in the SSR pattern were observed among individual plants of the same cultivar in four out of the eight local cultivars analysed. Several primer pairs (17%) amplified more than one locus. PMID- 12582691 TI - Comparative analysis of polymorphism and chromosomal location of tomato microsatellite markers isolated from different sources. AB - Previously isolated tomato ( Lycopersicon esculentum) microsatellite markers were mainly clustered in the centromeric heterochromatin and not located in euchromatic regions. To achieve a more-uniform distribution of microsatellite markers for genome mapping purposes, a set of tomato microsatellite markers containing dinucleotide simple sequence repeats were developed by screening genomic libraries enriched for single-copy sequences, and screening the tomato EST database. The tomato microsatellites isolated in these ways were characterized by combinations of different types of repeated motifs and they were polymorphic in a set of L. esculentum varieties detecting up to four alleles. A total of 20 markers were placed on the genetic map of tomato. Interestingly, all markers isolated from genomic libraries enriched for single-copy sequences by PstI-pre-digestion mapped into the centromeric regions. The majority of markers derived from EST sequences contained predominantly AT microsatellites and were located in euchromatic regions. PMID- 12582692 TI - Seed-specific, developmentally regulated genes of peanut. AB - Four cDNAs of seed-specific and developmentally regulated peanut ( Arachis hypogaeaL.) genes were identified by differential screening of a peanut-seed cDNA library using cDNA probes constructed from mRNAs isolated from immature and mature stages of the seed. Northern analysis, probed with the four cloned cDNAs, indicated that the genes represented by two cDNAs were expressed abundantly early in seed development, while another two were abundantly expressed later at the cell-expansion stages of seed development. These four genes did not show expression in roots, pegs or leaves. However, one of the early expressed genes was seed coat-specific. One of the clones, Psc11, had significant sequence similarity to subtilisin-like genes in Arabidopsis and soybean. Clones Psc32 and Psc33 had significant similarity to the peanut allergen genes Ara h II and Ara h 6, respectively. The sequence of clone Psc12 was unique and did not show significant similarity to any sequence in the databases. One of the four seed specific clones showed restriction fragment length polymorphism (RFLP) among peanut lines representing the four peanut botanical varieties. These findings indicate that polymorphism exists in peanut seed-storage genes. This contrasts with other genes previously used for genetic mapping of cultivated peanut. PMID- 12582693 TI - Extremely elongated tomato fruit controlled by four quantitative trait loci with epistatic interactions. AB - Cultivated tomato ( Lycopersicon esculentum) encompass a wide range of fruit shape and size variants. This variation can be used to genetically dissect the molecular basis of ovary and fruit morphology. The cultivar Long John displays an extremely elongated fruit phenotype, while the wild relative Lycopersicon pimpinellifolium LA1589 produces fruit that are nearly perfect spheres, typical of wild tomatoes. Quantitative trait mapping of an F2 population between Long John and LA1589 revealed four fruit shape QTLs, located on chromosomes 2, 3, 7 and 11. The primary role of the fruit shape QTL located on chromosome 7, ljfs7, is to control pericarp elongation. The primary role of the fruit shape QTLs on chromosome 2, 3 and 11 ( ljfs2, ljfs3 and ljfs11, respectively) is to control pear shape, measured as the eccentricity index. QTL map position and the effect of the loci on fruit shape suggested that ljfs2 and ljfs7 are allelic to the well studied fruit shape loci ovate and sun, respectively. ljfs3 and ljfs11 map near the previously identified, but less characterized, fruit shape loci fs3.2 and fs11.1, respectively. This result suggests that most of the variation in tomato fruit shape is controlled by a few major QTLs. Although eccentricity and pericarp elongation were largely controlled by independent growth processes, significant interactions were detected between all four fruit shape loci in the control of eccentricity. This indicates that the three eccentricity loci, ljfs2, ljfs3 and ljfs11, epistatically control the same developmental process, while ljfs7 had a pleiotropic effect on eccentricity. PMID- 12582694 TI - RFLP-facilitated investigation of the quantitative resistance of rice to brown planthopper ( Nilaparvata lugens). AB - Quantitative trait loci (QTLs), conferring quantitative resistance to rice brown planthopper (BPH), were investigated using 160 F(11) recombinant inbred lines (RILs) from the Lemont/Teqing cross, a complete RFLP map, and replicated phenotyping of seedbox inoculation. The paternal indica parent, Teqing, was more resistant to BPH than the maternal japonica parent, Lemont. The RILs showed transgressive segregation for resistance to BPH. Seven main-effect QTLs and many epistatic QTL pairs were identified and mapped on the 12 rice chromosomes. Collectively, the main-effect and epistatic QTLs accounted for over 70% of the total variation in damage scores. Teqing has the resistance allele at four main effect QTLs, and the Lemont allele resulted in resistance at the other three. Of the main-effect QTLs identified, QBphr5b was mapped to the vicinity of gl1, a major gene controlling leaf and stem pubescence. The Teqing allele controlling leaf and stem pubescence was associated with resistance, while the Lemont allele for glabrous stem and leaves was associated with susceptibility, indicating that this gene may have contributed to resistance through antixenosis. Similar to the reported BPH resistance genes, the other six detected main-effect QTLs were all mapped to regions where major disease resistance genes locate, suggesting they might have contributed either to antibiosis or tolerance. Our results indicated that marker-aided pyramiding of major resistance genes and QTLs should provide effective and stable control over this devastating pest. PMID- 12582695 TI - A physical map of a gene-dense region in soybean linkage group A2 near the black seed coat and Rhg (4) loci. AB - Soybean ( Glycine max L. Merrill) linkage group A2 contains a major resistance gene to the soybean cyst nematode ( Heterodera glycines Ichinohe) at the Rhg (4) locus near a gene encoding aspartokinase homoserine dehydrogenase (AK-HSDH) and also near the I locus affecting seed coat color. To identify clones related to this region of the genome, we used a PCR assay using primers designed from a gene encoding AK-HSDH to screen approximately 40,000 clones from a bacterial artificial chromosome (BAC) library constructed from genomic DNA of the susceptible cv. Williams 82. The identified BACs were screened with a second PCR assay using primers designed from DNA sequence associated with the I locus to confirm the location of the BACs. Only BAC Gm_ISb001_056_G02 (56G2) was positive for both assays. BAC 56G2 contains several genes previously associated with stress or defense response including genes with high sequence similarity to those encoding chalcone synthase, glucosyl-transferase, a heat-shock transcription factor, a membrane-associated salt-inducible protein, adenosyl homocysteinase, a protein kinase, and a G10-like protein. The map contributes to our understanding of the organization of the soybean genome and to the completion of a physical map of the soybean genome. In addition, the genes identified provide landmarks to identify BAC clones near the Rhg (4) locus in resistant soybean genomic libraries and provide a foundation for comparison of soybean cyst nematode resistant and susceptible DNA sequences in this region. PMID- 12582696 TI - Targeted isolation, sequence analysis, and physical mapping of nonTIR NBS-LRR genes in soybean. AB - Most cloned plant disease resistance genes (R-genes) code for proteins belonging to the nucleotide binding site (NBS) leucine-rich repeat (LRR) superfamily. NBS LRRs can be divided into two classes based on the presence of a TIR domain ( Toll and interleukin receptor-like sequence) or a coiled coil motif (nonTIR) in their N-terminus. We used conserved motifs specific to nonTIR-NBS-LRR sequences in a targeted PCR approach to generate nearly 50 genomic soybean sequences with strong homology to known resistance gene analogs (RGAs) of the nonTIR class. Phylogenetic analysis classified these sequences into four main subclasses. A representative clone from each subclass was used for genetic mapping, bacterial artificial chromosome (BAC) library screening, and construction of RGA-containing BAC contigs. Of the 14 RGAs that could be mapped genetically, 12 localized to a 25-cM region of soybean linkage group F already known to contain several classical disease resistance loci. A majority of the genomic region encompassing the RGAs was physically isolated in eight BAC contigs, together spanning more than 1 Mb of genomic sequence with at least 12 RGA copies. Phylogenetic and sequence analysis, together with genetic and physical mapping, provided insights into the genome organization and evolution of this large cluster of soybean RGAs. PMID- 12582697 TI - Linkage mapping in apomictic and sexual Kentucky bluegrass ( Poa pratensis L.) genotypes using a two way pseudo-testcross strategy based on AFLP and SAMPL markers. AB - The high versatility of the mode of reproduction and the retention of a pollen recognition system are the factors responsible for the extreme complexity of the genome in Poa pratensis L. Two genetic maps, one of an apomictic and one of a sexual genotype, were constructed using a two-way pseudo-testcross strategy and multiplex PCR-based molecular markers (AFLP and SAMPL). Due to the high ploidy level and the uncertainty of chromosome pairing-behavior at meiosis, only parent specific single-dose markers (SDMs) that segregated 1:1 in an F(1) mapping population (161 out of 299 SAMPLs, and 70 out of 275 AFLPs) were used for linkage analysis. A total of 41 paternal (33 SAMPLs and 8 AFLPs) and 47 maternal (33 SAMPLs and 14 AFLPs) SDMs, tested to be linked in coupling phase, were mapped to 7+7 linkage groups covering 367 and 338.4 cM, respectively. The comparison between the two marker systems revealed that SAMPL markers were statistically more efficient than AFLP ones in detecting parent-specific SDMs (75% vs 32.4%). There were no significant differences in the percentages of distorted marker alleles detected by the two marker systems (27.8% of SAMPLs vs 21.3% of AFLPs). The pairwise comparison of co-segregational groups for linkage detection between marker loci suggested that at least some of the P. pratensis chromosomes pair preferentially at meiosis-I. PMID- 12582698 TI - A male and hermaphrodite specific RAPD marker for papaya ( Carica papayaL.). AB - The random amplified polymorphic DNA (RAPD) technique was used to determine the sex of a dioecious species, Carica papayaL., with three sex types, male, female and hermaphrodite. A 450 bp marker fragment, named PSDM(Papaya Sex Determination Marker), exists in all male and hermaphrodite plants but not in the female plants so far analyzed. The DNA sequence of PSDM exhibited no significant similarity to previously reported sequences. A sequence-characterized amplified region (SCAR) marker, SCARps, was developed from PSDM to determine the sex of papaya. Southern hybridization, using PSDM as a probe, showed that PSDM exists in the male and hermaphrodite genomes, but not in the female genome. This result strongly suggests that PSDM is located on the chromosome region that is specific to the male and the hermaphrodite. SCARps is a suitable marker for the precise and rapid diagnosis of sex in papaya. PMID- 12582699 TI - Characterization of trinucleotide SSR motifs in wheat. AB - Length differences among trinucleotide-based microsatellite alleles can be more easily detected and frequently produce fewer "stutter bands" as compared to dinucleotide-based microsatellite markers. Our objective was to determine which trinucleotide motif(s) would be the most-polymorphic and abundant source of trinucleotide microsatellite markers in wheat ( Triticum aestivumL.). Four genomic libraries of cultivar 'Chinese Spring' were screened with nine trinucleotide probes. Based on the screening of 28550 clones, the occurrences of (CTT/GAA) (n), (GGA/CCT) (n), (TAA/ATT) (n), (CAA/GTT) (n), (GGT/CCA) (n), (CAT/GTA) (n), (CGA/GCT) (n), (CTA/GAT) (n), and (CGT/GCA) (n) repeats were estimated to be 5.4x10(4), 3.5x10(4), 3.2x10(4), 1.2x10(4), 6.3x10(3), 4.9x10(3), 4.5x10(3), 4.5x10(3) and 3.6x10(3), i.e., once every 293 kbp, 456 kbp, 500 kbp, 1.3 Mbp, 2.6 Mbp, 3.2 Mbp, 3.6 Mbp, 3.6 Mbp and 4.5 Mbp in the wheat genome, respectively. Of 236 clones selected for sequencing, 38 (93%) (TAA/ATT) (n), 30 (43%) (CTT/GAA) (n), 16 (59%) (CAA/GTT) (n), 3 (27%) (CAT/GTA) (n) and 2 (4%) (GGA/CCT) (n) clones contained microsatellites with eight or more perfect repeats. From these data, 29, 27 and 16 PCR primer sets were designed and tested to the (TAA/ATT) (n), (CTT/GAA) (n) and (CAA/GTT) (n) microsatellites, respectively. A total of 12 (41.4%) primers designed to (TAA/ATT) (n), four (14.8%) to (CTT/GAA) (n), and two (12.5%) to (CAA/GTT) (n) resulted in polymorphic markers. The results indicated that (TAA/ATT) (n) microsatellites would provide the most-abundant and the most-polymorphic source of trinucleotide microsatellite markers in wheat. PMID- 12582700 TI - Common loci underlie field resistance to soybean sudden death syndrome in Forrest, Pyramid, Essex, and Douglas. AB - Soybean [ Glycine max (L.) Merr.] sudden death syndrome (SDS) caused by Fusarium solani f. sp. glycines results in severe yield losses. Resistant cultivars offer the most-effective protection against yield losses but resistant cultivars such as 'Forrest' and 'Pyramid' vary in the nature of their response to SDS. Loci underlying SDS resistance in 'Essex' x Forrest are well defined. Our objectives were to identify and characterize loci and alleles that underlie field resistance to SDS in Pyramidx'Douglas'. SDS disease incidence and disease severity were determined in replicated field trials in six environments over 4 years. One hundred and twelve polymorphic DNA markers were compared with SDS disease response among 90 recombinant inbred lines from the cross PyramidxDouglas. Two quantitative trait loci (QTLs) for resistance to SDS derived their beneficial alleles from Pyramid, identified on linkage group G by BARC Satt163 (261-bp allele, P=0.0005, R(2)=16.0%) and linkage group N by BARC-Satt080 (230-bp allele, P=0.0009, R(2)=15.6%). Beneficial alleles of both QTLs were previously identified in Forrest. A QTL for re- sistance to SDS on linkage group C2 identified by BARC-Satt307 (292-bp allele, P=0.0008, R(2)=13.6%) derived the beneficial allele from Douglas. A beneficial allele of this QTL was previously identified in Essex. Recombinant inbred lines that carry the beneficial alleles for all three QTLs for resistance to SDS were significantly ( P0.97, the P values were 0.000, and sensitivity was 94%, specificity 97%, predictive value for resistance >/=92%, predictive value for susceptibility 97%, and test efficiency 97%. According to ROC analysis, the microplate Alamar blue assay is a reliable method for determination of drug-susceptibility. Rapidity and cost efficiency are two additional qualities that make this test an excellent alternative for the drug susceptibility testing of Mycobacterium tuberculosis. The ROC curve analysis is a robust statistical approach for evaluating the performance of new quantitative methods for determination of drug sensitivity of Mycobacterium tuberculosis isolates. PMID- 12582741 TI - Use of the E test to assess synergy of antibiotic combinations against isolates of Burkholderia cepacia-complex from patients with cystic fibrosis. AB - Treatment of Burkholderia cepacia-complex infections in cystic fibrosis patients is problematic, since the microorganism is often resistant to most antimicrobial agents. In this study, the Epsilometer test, or E test, was used to assess the activity of antimicrobial combinations against Burkholderia cepacia-complex. In a preliminary evaluation, the E test was compared to the checkerboard method using 10 test organisms. Synergy testing by the E test was then performed on 131 clinical isolates of Burkholderia cepacia-complex using various combinations of antimicrobial agents. Agreement between the E test and the checkerboard method was 90%. The rate of resistance to individual agents ranged from 48% for meropenem to 100% for tobramycin, chloramphenicol, and rifampin. In 71.6%, 15.6%, and 12.6% of the test evaluations performed, the combinations tested resulted in additivity/indifference, synergism, and antagonism, respectively. The highest rates of synergy were observed with combinations of ciprofloxacin-piperacillin (44%), rifampin-ceftazidime (33%), chloramphenicol-ceftazidime (22%), cotrimoxazole-piperacillin/tazobactam (22%), and ciprofloxacin-ceftazidime (21%). Rates of antagonism for cotrimoxazole and chloramphenicol in combination with beta-lactam agents were higher than those observed for ciprofloxacin plus beta lactam agents. These results suggest that the E test is a valuable and practical method to be considered for improving the identification of possible therapeutic options in cystic fibrosis patients infected with organisms belonging to the Burkholderia cepacia-complex. PMID- 12582742 TI - Comparison of the sodium dodecyl sulfate-sodium hydroxide specimen processing method with the C18-carboxypropylbetaine specimen processing method using the MB/BacT liquid culture system. AB - The ability of physicians to diagnose tuberculosis is impacted by the use of smear and culture techniques combined with specimen processing methods. The objective of this study was to evaluate the effects of specimen processing on smear and culture sensitivity by comparing the specimen processing method that uses C(18)-carboxypropylbetaine with the method that combines sodium dodecyl sulfate and sodium hydroxide. A total of 1,201 specimens were entered into this study. Specimens were split approximately equally such that one-half of each specimen was processed with sodium dodecyl sulfate-sodium hydroxide, while the other half was processed with C(18)-carboxypropylbetaine. All sediments were subjected to acid-fast staining and then analyzed using the MB/BacT liquid culture system (bioMerieux, France) and solid media. The sensitivity of smear following processing with sodium dodecyl sulfate-sodium hydroxide and C(18) carboxypropylbetaine was 61.2% and 58.6% (P>0.05), respectively, while the specificities were identical (99.7%). The sensitivity of culture was 84.2% and 96.1% (P<0.05), respectively. The time to detection in the MB/BacT liquid culture system was 13.2+/-5.6 and 15.0+/-8.8 days (P>0.05), respectively, and 20.0+/-7.6 and 15.7+/-8.9 days (P<0.05), respectively, on solid media. The contamination rates in the MB/BacT system were 0.8% and 8.7%, respectively, whereas the contamination rates on solid media were 2.6% and 4.3%, respectively. C(18) carboxypropylbetaine specimen processing was less labor-intensive than sodium dodecyl sulfate-sodium hydroxide processing and improved the ability of laboratory staff to detect the presence of mycobacteria by culture. PMID- 12582743 TI - Duration of antifungal treatment and development of delayed complications in patients with candidaemia. AB - The optimal duration of antifungal treatment of candidaemia is unknown. Prolonged treatment has been advocated to prevent late-onset complications caused by metastatic infectious foci. To evaluate the relationship between duration of antifungal therapy and the development of delayed complications in patients with candidaemia, a retrospective descriptive study was performed in a tertiary care centre. The study included 180 consecutive patients with candidaemia identified by at least one positive blood culture. Development of late-onset complications of candidaemia during long-term follow-up was monitored. Of the patients treated with antifungal agents, 55% completed treatment without apparent complications, 33% died during therapy, 12% developed disseminated disease and one patient continues to receive ongoing treatment. Of the 81 patients who completed treatment as scheduled, 25% received treatment for less than 2 weeks, 31% for 2-4 weeks and 38% for more than 4 weeks; duration of therapy was unknown in 6%. Only three (2%) patients developed delayed complications; in these patients, the duration of antifungal treatment had been 3 weeks, 5 weeks and 22 weeks, respectively. In conclusion, despite theoretical concerns, there seems to be no correlation between the duration of antifungal treatment and the development of late complications in patients with candidaemia. This suggests that treatment of 2 weeks or less may be sufficient, provided the initial response to therapy is favourable. PMID- 12582744 TI - A case of tuberculous sinusitis without concomitant pulmonary disease. AB - This case report highlights the difficulty of diagnosing tuberculous sinusitis in the absence of pulmonary foci. Although extrapulmonary localisations of tuberculosis are rare in immunocompetent patients, it is important to consider this diagnosis, since therapeutic delay usually results in an unfavourable outcome. Acid-fast bacilli are sometimes difficult to detect in pathological specimens. Consequently, the diagnosis is usually based on the following criteria: (i) the absence of clinical response to usual antibiotics, (ii) the presence of caseous granulomatous inflammatory lesion on histopathology, and (iii) identification of Mycobacterium tuberculosis by the polymerase chain reaction assay confirmed by bacteriological culture. The diagnosis of tuberculosis is finally confirmed by the efficacy of antituberculous treatment. The differential diagnosis is Wegener's disease. PMID- 12582746 TI - Rapid and standardized detection of Chlamydia pneumoniae using LightCycler real time fluorescence PCR. AB - In order to meet the need of many microbiological laboratories for a standardized system for detecting Chlamydia pneumoniae in respiratory specimens, a hybridization probe-based LightCycler (Roche Diagnostics, Germany) PCR assay was developed. The assay's analytical sensitivity and specificity were evaluated according to the recommendations of the Centers for Disease Control and Prevention (USA) and Laboratory Centre for Disease Control (Canada). Seventy-four bacterial species other than Chlamydia pneumoniae, including strains of Chlamydia trachomatis, Chlamydia psittaci, and Chlamydia pecorum, tested negative. Six of six representative Chlamydia pneumoniae strains tested positive. An analytical sensitivity of 1 inclusion forming unit per ml of bronchoalveolar lavage fluid, corresponding to 0.02 inclusion forming units per PCR reaction, was observed. The assay showed 100% specificity and sensitivity for Chlamydia pneumoniae when testing DNA preparations from 12 specimens of patients with known pulmonary Chlamydia pneumoniae infection and from 78 specimens of patients with respiratory tract disease of other origin. The newly developed LightCycler assay may contribute to the urgently needed standardization of laboratory diagnosis of Chlamydia pneumoniae. PMID- 12582745 TI - Relapsing cutaneous alternariosis in a kidney transplant recipient cured with liposomal amphotericin B. AB - An immunosuppressed patient who presented with unusual clinical signs of cutaneous alternariosis, including papular, nodular and verrucous lesions of the forearms, is reported. In spite of continuous treatment with oral itraconazole for 6 months, a large, progressive, necrotic ulcer appeared on the patient's left leg. Liposomal amphotericin B was then administered (total dose, 750 mg) with excellent clinical results. PMID- 12582747 TI - Molecular epidemiology of oxacillin-resistant Staphylococcus aureus in Brooklyn, New York. AB - Although oxacillin-resistant Staphylococcus aureus (ORSA) are well-established pathogens in many hospitals, the impact of infection control measures on the clonal distribution of ORSA is poorly defined. A citywide surveillance study of Staphylococcus aureus in Brooklyn, New York revealed that 36% of isolates were ORSA. Molecular typing showed that one strain (Cluster A) accounted for more than half of the isolates and was present in all 15 hospitals. In one hospital, a distinct strain (Cluster B) accounted for 20% of ORSA isolates. Infection control measures in this hospital significantly decreased the percentage of clinical isolates that belonged to Cluster B, but did not have an effect on the strain endemic to the city (Cluster A). Regional infection control strategies may need to be developed to limit the spread of the ORSA clone endemic to this area. PMID- 12582749 TI - First isolation of virulent Yersinia enterocolitica O8, biotype 1B in Germany. PMID- 12582750 TI - Long-term CD4+ T-cell response in HIV-1-infected patients continuing combination antiretroviral therapy despite high-level viremia and drug resistance. PMID- 12582748 TI - Disseminated infection due to Cylindrocarpon (Fusarium) lichenicola in a neutropenic patient with acute leukaemia: report of a case and review of the literature. AB - Described here is an unusual case of disseminated Cylindrocarpon lichenicola (Fusarium lichenicola) infection originating from a toenail lesion of a neutropenic woman with cellulitis of the foot and underlying acute leukaemia. A computed tomography scan of the chest showed multiple, ill-defined, nodular infiltrates with alveolar consolidation. The fungus was isolated from both the nail and the skin of the infected toe. Susceptibility testing revealed low minimum inhibitory concentrations for amphotericin B (0.78 micro g/ml) and voriconazole (1.56 micro g/ml) and high minimum inhibitory concentrations (>100 micro g/ml) for fluconazole, ketoconazole and itraconazole. The infection resolved after treatment with a total dose of 1 g of amphotericin B followed by oral itraconazole and bone marrow regeneration. PMID- 12582751 TI - Highly resistant Corynebacterium macginleyi as cause of intravenous catheter related infection. PMID- 12582752 TI - Construct validity: experts and novices performing on the Xitact LS500 laparoscopy simulator. AB - BACKGROUND: This study was undertaken to establish construct validity of the Xitact LS500, a virtual reality laparoscopic cholecystectomy simulator. The primary research statement is: "The clip-and-cut task on the Xitact LS500 virtual reality laparoscopic cholecystectomy simulator mimics the surgical procedure of the clipping and cutting of the cystic duct and artery during the laparoscopic cholecystectomy adequately." METHODS: According to the level of experience of the surgeon, an "expert group opinion" was formed resulting from 37 surgeons having performed over 100 laparoscopic cholecystectomies, and a "novice group opinion" was formed resulting of 37 surgeons having no experience at all with performing laparoscopic cholecystectomy. Participants received an instructed hands-on "tour" on the Xitact simulator and performed three formal simulation runs. RESULTS: The "novice group" is younger and more surgeons are female. Performance scores in the "expert" group are significantly higher on the second (p value 0.011) and third (p value 0.005) run, compared to the novices' scores. Experts are significantly faster on completion of all three runs. There is an increase in score over runs in both groups, which is statistically significant in the "expert group." Less than one-third of surgeons in either group are able to correctly predict their performance score as generated by Xitact. Both "experts" and "novices" feel it is useful to train with Xitact LS500 in the surgical curriculum. CONCLUSIONS: Three hypotheses, formulated to operationalize the primary research statement, could be answered affirmatively. Although further validation studies are needed, the Xitact LS500 simulator seems to be able to discriminate between expert and novice surgeons in this research setting, and thus the construct for this setting is considered to be valid. PMID- 12582754 TI - A prospective randomized trial on comparison of low-pressure (LP) and standard pressure (SP) pneumoperitoneum for laparoscopic cholecystectomy. AB - AIM: This study aimed to investigate the advantages and disadvantages of LP (7 mmHg) in comparison to SP (12 mm Hg) pneumoperitoneum in a prospective randomized clinical trial. MATERIALS AND METHODS: 148 consecutive patients qualified for laparoscopic cholecystectomy (LC) due to uncomplicated symptomatic gallstones were randomized to either SPLC or LPLC. All the procedures were performed by the same experienced team of surgeons. The statistical analysis included sex, mean age, body mass index, ASA grade, operative time, complication rate, conversion rate, postoperative pain assessed by the Visual Analogue Scale of Pain (VAS) including the incidence of shoulder-tip pain, postoperative hospital stay, recovery time, and the quality of life (QOL) within 7 days following the operation. p <0.05 was considered as indicative of significance. RESULTS: Neither conversion to an open procedure nor major complications occurred in either group. The operative time was similar in both groups (LP 55.7 +/- 8.6 min vs SP 51.9 +/- 8.3 min). The mean postoperative pain score was 6.18 +/- 3.48 lower after LP than SPLC and the difference amounted to 22.2% (p <0.005). The incidence of shoulder tip pain was 2.1 times lower after LP than SPLC (p <0.05). QOL within 7 days following the operation was remarkably better after LPLC than after SPLC (p <0.01). CONCLUSIONS: LP pneumoperitoneum is superior to SP pneumoperitoneum in terms of lower postoperative pain, a lower incidence of shoulder-tip pain, and a better QOL within 5 days following the operation. LP should be used for LC in cases of uncomplicated symptomatic gallstones as a recommended procedure as long as an adequate exposure is obtained with this technique. PMID- 12582753 TI - Changing trends in the management of gallstone disease. AB - BACKGROUND: Day case cholecystectomy is increasingly becoming a management option for elective cases while "same admission" cholecystectomy is now considered a favorable option in the treatment of acute cholecystitis. To assess the advent of these changes in our surgical practice, a retrospective analysis of our experience is presented. METHODS: All patients undergoing cholecystectomy between January 2000 and January 2001 were analyzed according to admission status, operation type, conversion rate, complications, and nonsurgical intervention. RESULTS: 156 patients underwent cholecystectomy and 152 charts were retrieved. Laparoscopic cholecystectomy was performed on 95% of patients with a conversion rate of 9%. Morbidity for the series was 12.5%, including one common bile duct injury (0.6%). Day case and acute cholecystectomy comprised 67% of our cholecystectomy practice. CONCLUSIONS: Our findings suggest that there is an increasing trend toward shortening the hospital stay of patients undergoing laparoscopic cholecystectomy. This does not appear to have had a deleterious effect on outcome. PMID- 12582755 TI - Endoscopic ablation of Barrett's esophagus using argon plasma coagulation (APC) following surgical laparoscopic fundoplication. AB - BACKGROUND: Barrett's esopagus (BE) is considered a risk factor for the development of esophageal carcinoma. Recently, partial restoration of squamous mucosa after ablation of BE with endoscopic techniques has been described. METHODS: From November 1996 to November 1999, 23 patients with histologically proven BE have been treated by endoscopic argon plasma coagulation (APC) following suppression of gastro-esophageal reflux by laparoscopic fundoplication. Histological follow-up after completed ablation ranged from 16 to 45 months (mean, 31.9 months). RESULTS: Histologically, complete squamous reepithelialization was observed in 20/23 patients, whereas a regrowth of a mixed squamous and gastric type mucosa was observed in 1 patient. Small islands of intestinal metaplasia were observed under the neosquamous epithelium in two patients (9%) during follow-up. CONCLUSION: The success rate of APC ablation following laparoscopic antireflux surgery in our series may be as high as 91%. Nevertheless, small islands of intestinal metaplasia under the new squamous epithelium may persist in some patients. In these circumstances, the authors recommend that endoscopic ablation of BE should be confined to controlled clinical trials. PMID- 12582756 TI - Live donor nephrectomy and return to work: does the operative technique matter? AB - BACKGROUND: Several studies report an earlier return to work after minimal invasive kidney donation compared to open donor nephrectomy. However, this variation in outcome might be influenced by other factors than the surgical technique used, such as the advice given by the physician regarding return to work. In this study, we compare the absence from work after open (ODN), laparoscopic (LDN), and hand-assisted donor nephrectomy (HA) performed in the Netherlands, in relation to the advice given. METHODS: Questionnaires containing questions about return to work or return to daily activities were sent to 78 donors from three hospitals. In the HA and ODN hospitals, advice on full return to work was 3 months. In contrast, advice given in the LDN hospital was 6 weeks. RESULTS: After LDN, donors resumed their work after 6 weeks, 5 weeks faster compared to ODN (p = 0.002) and HA (p <0.001). Complete return to work occurred 9 weeks sooner in the LDN group compared to the ODN and HA groups (both p <0.001). In the unemployed group, there was no significant difference in length until full return to daily activities. CONCLUSION: Return to work is influenced by the advice on return to work given by the physician as well as the morbidity associated with the surgical approach. PMID- 12582757 TI - Minimal invasive approach of gastric and esophageal mobilization in total pharyngolaryngoesophagectomy: total laparoscopic and hand-assisted laparoscopic technique. AB - BACKGROUND: To evaluate early results in total pharyngolaryngoesophagectomy (PLE) by minimally invasive approaches for patients suffered from pharyngoesophageal tumor. METHODS: Between April 1998 and September 2001, 12 consecutive patients underwent either total laparoscopic (n = 9) or hand-assisted laparoscopic (n = 3) gastric mobilization plus transhiatal esophageal resection in total PLE. The operative data and postoperative outcomes were evaluated. RESULTS: Total PLE by minimally invasive approach was successfully performed in 11 patients, and 1 patient required conversion due to uncontrolled bleeding. The median total operative time was 8.5 h (range, 5-11 h) and the abdominal laparoscopic stage usually took less than 4 h. The median time for extubation was 2 days (range, 1-4 days) and the median ICU stay was 2 days (range, 1-20 days). There was no 30-day mortality, and major complications occurred in 5 patients (42%). CONCLUSION: Minimally invasive PLE is a feasible and safe alternative to conventional open surgery for patients with pharyngoesophageal carcinoma. PMID- 12582758 TI - Laparoscopy in the last decade of the millennium: have we really improved? AB - BACKGROUND: The aims of the study were to evaluate the evolution of laparoscopic surgery during the past decade in terms of variations in the quality (complexity) of the procedures performed and of modifications in patient outcome. METHODS: A retrospective analysis was performed of 3022 consecutive patients undergoing 99 different laparoscopic procedures at a center specialized in laparoscopic abdominal surgery. All the procedures were classified according to three classes of complexity. Results relating to the first 1511 patients were compared to those of the last 1511 patients. RESULTS: In the second group, medium- to high-class complexity procedures significantly increased, conversion rate was higher only for straightforward procedures, duration of low- to medium-class complexity procedures decreased, only the rate of slight complications increased, and mean postoperative hospital stay was longer. Frequency of conversion in medium- to high-class complexity procedures and severe complications was not different in the two periods. CONCLUSIONS: The quality of laparoscopic surgery has improved during the past decade, with no increase in the frequency of conversion or of major complications. PMID- 12582759 TI - Can tonic accommodation predict surgical performance? AB - BACKGROUND: Pilots undergo many visual tests for both selection and assessment, and we know that there are many similarities between pilots and surgeons. Hence, it would not be unreasonable to bring similar visual tests into surgery. Tonic accommodation (TA) is a stable parameter that is adopted by the eye in the absence of any stimulation. Over recent years, surgery has undergone change from traditional open surgery to minimally invasive procedures, bringing many advantages. However, not every surgeon has the ability to perform under conditions where the operative field is represented on a flat monitor. METHOD: We determined the TA values in medical students and then correlated this with their performance on a virtual reality surgical simulator. RESULTS: We found that TA values predicted the number of errors made with the dominant hand, accounting for 27% of the variance. CONCLUSION: The data suggest that TA may play a role in the individual differences that are noted when surgeons perform laparoscopic surgery. Further studies are needed to evaluate the exact role of TA in surgical performance. PMID- 12582760 TI - Our experience with 250 laparoscopic adjustable silicone gastric bandings. AB - BACKGROUND: In 1999, after having performed more than 2500 open bariatric surgery procedures, we began to conduct the laparoscopic adjustable silicone gastric banding (LASGB) procedure. The purpose of this report is to present the results of our first 250 operations using this technique. PATIENTS AND METHODS: The 250 patients included 212 females and 38 males. The mean age was 36 years (range, 16 62). The mean BMI was 44 (range, 35-76). Patients were selected in accordance with standard criteria. The mean follow-up time was 18 months (range, 3-24). RESULTS: Two hundred and forty procedures were completed by laparoscopy; conversion to open surgery was required in 10 cases. The mean operating time was 50 min (range, 20-145). There was no mortality. Perioperative complications included two cases of gastric perforation. Late complications included 13 cases of band slippage (5%), of which three involved penetration to the stomach, five cases of port disconnection (2%), and four cases of port site infection (1.5%). The mean weight loss at 12 months was 70% of the original excess weight. CONCLUSION: Most patients prefer laparoscopy as the surgical technique in the treatment of morbid obesity. Our initial experience with 250 operations confirms that this approach is as good as open surgery. The long learning curve is steep, requiring at least 100 cases to guarantee low complication rates. PMID- 12582761 TI - Significant symptomatic relief after transoral endoscopic staple-assisted treatment of Zenker's diverticulum. AB - BACKGROUND: Zenker's diverticulum is associated with characteristic symptoms of progressive dysphagia and regurgitation. As most patients are elderly, the perioperative risk is usually high. We report our clinical experience with the transoral endoscopic staple-assisted method, including a thorough assessment of the symptomatic relief achieved by the procedure. METHODS: 46 consecutive patients (29 m, 17 f) with a median age of 61 years (range, 37-96 years) were treated between 1997 and 2002. The symptoms and their frequency were registered. The follow-up consisted of clinical and radiographic investigations. RESULTS: The median size of the diverticulum was 4 cm (range, 2-12 cm). Transoral treatment was successful in 39 patients; in 7 cases (15.2%) a switch to open surgery was required. The median operating time was 30 min (range, 10-150 min). Mortality rate was nil, while morbidity was 7.7%. The median duration of the postoperative hospital stay was 5 days (range, 1-65 days). After a median follow-up of 11 months (range, 1-40 months), 5 patients had been reoperated on endoscopically due to clinical recurrence. Clinical symptoms were significantly reduced (dysphagia of liquids p 80% of the patients, corresponding to a loss of 15 body mass index (BMI) units in morbidly obese patients and 20 BMI units in superobese patients. In the vast majority of patients, comorbidities improved or disappeared over time and quality of life improved. CONCLUSIONS: Laparoscopic Roux-en-Y gastric bypass is feasible, but it is a very complex operation. Indeed, it is associated with a long and steep learning curve, as reflected in the high number of major complications among our first 70 patients. The learning curve probably includes between 100 and 150 patients. With increasing experience, the morbidity rate becomes more acceptable and comparable to that of open RYGBP. The results in terms of weight loss and correction of comorbidities are similar to those obtained after open surgery, at least in the short term. However, only surgeons with extensive experience in advanced laparoscopic as well as bariatric surgery should attempt this procedure. PMID- 12582768 TI - Laparoscopic herniorrhaphy in children. AB - BACKGROUND: We report our clinical experience with 403 inguinal hernias in 279 children. They were treated via a purely laparoscopic approach using 2-mm instruments, obviating the need for a groin incision. METHODS: Laparoscopic herniorrhaphy was performed in children ages 4 days to 15 years. A 5-mm laparoscope was inserted through the umbilicus, and two 2-mm needle holders were inserted through the inferolateral abdominal wall. The open inner inguinal rings were closed by placing Z-sutures of monofilamentous nonabsorbable material. RESULTS: The mean operating time was 14 min for unilateral hernias and 21 min for bilateral hernias. We found 3.9% direct hernias. Hydroceles occurred in 1.7% of patients, testicular atrophy was noted in one patient, and no hernia was found in 2.3%. In girls with inguinal hernias, a contralateral asymptomatic patent processus vaginalis (PPV) was found in 45.2%, regardless of whether the hernia was on the right or the left side. In boys with inguinal hernias, contralateral PPVs were found on the right side in 21.9% and on the left in 8%. There were no major complications. One conversion to an open procedure was necessary because of a dilated bowel. The mean follow-up period was 23 months. There were 2.7% hernia recurrences; this rate was slightly higher than that seen with the open technique. The incidence of direct inguinal hernias was higher than has been previously reported. CONCLUSIONS: Laparoscopic herniorrhaphy allows the surgeon to identify the type of defect and proceed with immediate treatment. This technique is safe, reproducible, and technically easy for experienced laparoscopists. Bilaterality is of no concern. The cosmetic results are excellent; and in patients with recurrence of a hernia, this procedure is preferable to the open technique. PMID- 12582769 TI - Comparison of laparoscopic skills performance between standard instruments and two surgical robotic systems. AB - BACKGROUND: Our objective was to compare the performance of laparoscopic tasks by surgeons using standard laparoscopic instruments and two surgical robotic systems. METHODS: Eighteen surgeons performed tasks in a training box using three different instrument systems: standard laparoscopic instruments, the Zeus Robotic Surgical System, and the da Vinci Surgical System. Basic tasks included running a 100-cm rope, placing beads onto pins, and dropping cotton peanuts into cylinders; fine tasks included intracorporeal knot tying and running stitches with 4-0, 6-0, and 7-0 sutures. Time (in seconds) required and precision (number of errors) in performing each task were recorded. Analysis of variance with pair-wise comparisons using the Bonferroni method and Friedman's nonparametric test were used for statistical analysis. RESULTS: Standard instruments performed significantly faster than either robotic system on the rope and bead tasks (p <0.05), whereas da Vinci performed significantly faster than Zeus in all three basic tasks (p <0.05). No significant difference in precision was found between standard instruments and the robotic systems on any of the basic tasks. Knot tying and running-suture time were similar between standard instruments and da Vinci, which were significantly faster than Zeus (p <0.05) for all suture sizes. The robotic systems were similar in precision for fine suturing tasks and were significantly more precise in knot tying (Zeus and da Vinci) and running sutures (da Vinci) than standard instruments (p <0.05). CONCLUSIONS: Basic laparoscopic task performance is generally faster and as precise using standard instruments compared to either robotic system. In performing fine tasks, neither robotic system is faster than standard instruments, although they may offer some advantage in precision. PMID- 12582770 TI - Lower esophageal sphincter injection of a biocompatible polymer: accuracy of implantation assessed by esophagectomy. AB - BACKGROUND: Endoscopic lower esophageal sphincter (LES) implantation of a biocompatible polymer is undergoing clinical trial as an alternative to pharmacologic and laparoscopic treatments for gastroesophageal reflux disease. The safety and efficacy of LES augmentation depend on accurate placement of the implant into the wall of the esophagus. To date, no study has demonstrated the prevalence and location of the intended implant. METHODS: The study group consisted of nine patients with underlying esophageal disease severe enough to warrant esophagectomy. Three or four implants of 1 or 2 cc of Enteryx (a biocompatible ethylene-vinyl alcohol copolymer dissolved in dimethyl sulfoxide with micronized tantalum as a radiopaque marker) were placed at the squamocolumnar junction of each patient via a 4-mm, 23-gauge needle under endoscopic guidance. Fluoroscopy was utilized in all patients to facilitate endoscopic placement. Outcome measures included the prevalence and location of successful implantation into the wall of the esophagus. RESULTS: Thirty of 34 implants (88%) were successfully placed into the wall of the esophagus. The remaining 4 were found lying subserosally or attached to the exterior of the gastroesophageal junction (GEJ). Fluoroscopically, the implants often tended to coalesce, forming arcs or a ring around the GEJ. Histologic examination revealed implantation into the deep submucosa contiguous with the circular muscle and within the muscularis propria in all patients, with implants occasionally extending into the subserosa. There were no untoward reactions identified. CONCLUSIONS: Endoscopically directed implantation of a biocompatible polymer into the esophageal wall can be accomplished with a high degree of accuracy. Injection via a 4-mm needle results in the placement of material along and within the muscular layers of the esophagus. PMID- 12582771 TI - Evaluating minimally invasive surgery training using low-cost mechanical simulations. AB - BACKGROUND: The goal of this study was to develop, test, and validate the efficacy of inexpensive mechanical minimally invasive surgery (MIS) model simulations for training faculty, residents, and medical students. We sought to demonstrate that trained and experienced MIS surgeon raters could reliably rate the MIS skills acquired during these simulations. METHODS: We developed three renewable models that represent difficult or challenging segments of laparoscopic procedures; laparoscopic appendectomy (LA), laparoscopic cholecystectomy (LC), and laparoscopic inguinal hernia (LH). We videotaped 10 students, 12 surgical residents, and 1 surgeon receiving training on each of the models and again during their posttraining evaluation session. Five MIS surgeons then assessed the evaluation session performance. For each simulation, we asked them to rate overall competence (COM) and four skills: clinical judgment (respect for tissue) (CJ), dexterity (economy of movement) (DEX), serial/simultaneous complexity (SSC), and spatial orientation (SO). We computed intraclass correlation (ICC) coefficients to determine the extent of agreement (i.e., reliability) among ratings. RESULTS: We obtained ICC values of 0.74, 0.84, and 0.81 for COM ratings on LH, LC, and LA, respectively. We also obtained the following ICC values for the same three models: CJ, 0.75, 0.83, and 0.89; DEX, 0.88, 0.86, and 0.89; SSC, 0.82, 0.82, and 0.82; and SO, 0.86, 0.86, and 0.87, respectively. CONCLUSIONS: We obtained very high reliability of performance ratings for competence and surgical skills using a mechanical simulator. Typically, faculty evaluations of residents in the operating room are much less reliable. In contrast, when faculty members observe residents in a controlled, standardized environment, their ratings can be very reliable. PMID- 12582772 TI - Laparoscopic management of complications following laparoscopic Roux-en-Y gastric bypass for morbid obesity. AB - BACKGROUND: We reviewed our experience with complications following laparoscopic Roux-en-Y gastric bypass (LRYGB) that were managed laparoscopically. METHODS: A total of 246 consecutive morbidly obese patients (mean body mass index, 50.9 kg/m2) underwent LRYGB by three surgeons at two institutions. All patients met National Institutes of Health criteria for surgical treatment of morbid obesity. Patients were followed prospectively. RESULTS: A total of 62 patients (25.2%) developed 64 complications, 34 of which (13.8%) required a surgical intervention. Twenty-seven of the 34 procedures were performed laparoscopically. Gastrojejunostomy stricture was the most common complication (8.9%), followed by intestinal obstruction (7.3%) and gastrointestinal bleeding (4%). The intestinal obstruction was secondary to adhesions (n = 6), internal hernia at the level of the transverse mesocolon (n = 3), jejunojejunostomy stricture (n = 3), and cicatrix around the Roux limb at the level of the transverse mesocolon (n = 3). Other complications included gastrojejunostomy leak (1.6%), symptomatic gallstone disease (2.8%), and gastric remnant perforation (0.8%). One patient underwent a negative laparoscopy to rule out anastomotic leak. There were 3 deaths in this series of patients, 2 attributable to anastomotic leak. CONCLUSIONS: A variety of complications can present after LRYGB. Laparoscopy is an excellent technique to treat these complications. PMID- 12582773 TI - Laparoscopic mesh cruroplasty for large paraesophageal hernias. AB - BACKGROUND: Previous studies have shown that surgical repair of paraesophageal hernias is associated with a high recurrence rate, especially when a laparoscopic approach is used. Anatomic recurrence due to crura breakdown is a primary etiology, which has led us to employ prosthetic mesh reinforcement of large hiatal defects (> 5 cm) since 1996. We discuss the evolution of this approach and describe our current technique with limited outcomes in 52 consecutive patients during a 5-year period. METHODS: There were 31 males and 21 females, with a mean age of 57 years (range, 32-77 years) with symptomatic reflux and endoscopic or radiologic evidence of a large paraesophageal hernia. Utilizing a laparoscopic approach, the contents of the sac were reduced and the crura approximated with permanent interrupted sutures and a prosthetic mesh was secured over the repair as an on-lay reinforcement buttress. A Nissen (42) or Tilley (9) fundoplication was performed in all but 1 patient, and 18 patients (34%) required a wedge collis gastroplasty. Fifteen patients (28%) had a previous unsuccessful antireflux operation. RESULTS: There was no perioperative morbidity or mortality. Follow-up averaged 25 months (range, 7-60 months). Postoperative gastroscopy or barium swallow have been performed in 27 patients to date, with 11 experiencing foregut symptoms. There has been 1 recurrence (1.9%) and no prosthetic erosion. CONCLUSION: Early results suggest that prosthetic mesh cruroplasty may be effective in reducing recurrence after laparoscopic repair of large paraesophageal hernias, but long-term follow-up is required in all patients to determine the true incidence of anatomic recurrence and prosthetic erosion. PMID- 12582774 TI - Endosonography of upper gastrointestinal tract cancer on demand using miniprobes or endoscopic ultrasound. AB - BACKGROUND: The aim of this study was to investigate whether endosonography on demand with miniprobes and conventional endoscopic ultrasound improves the accuracy of endosonographic staging of upper gastrointestinal tract cancer. METHODS: Altogether, 173 patients underwent endoscopic ultrasonography for preoperative staging of esophageal (n = 63) or gastric cancer (n = 110). Depending on the endoscopic appearance (i.e., size and growth pattern), tumors were examined with a linear-array echoendoscope (7.5 MHz) or with high-resolution miniprobes (12.5 MHz). The results of preoperative staging were correlated with histopathology of the resection specimen. RESULTS: The overall accuracy of miniprobe ultrasonography and endoscopic ultrasound in assessing the infiltration depth of upper gastrointestinal cancer was 87% and 81%, respectively. Miniprobe ultrasonography was superior to conventional endoscopic ultrasound in the staging of early cancers, particularly T1 tumors (accuracy, 81% vs 56%). The combined accuracy of both techniques for all tumor stages was 82%. Correct diagnosis of lymph node involvement was obtained with miniprobe ultrasonography or endoscopic ultrasound in 76% and 71% of the cases, respectively. The combined accuracy in assessing the lymph node status was 73% (sensitivity, 68%; specificity, 81%). CONCLUSIONS: Endosonography on demand using either miniprobes or conventional endoscopic ultrasound may result in more effective and less invasive staging of esophageal and gastric cancer. Selective use of high-resolution miniprobes and conventional endoscopic ultrasound offers accurate staging of all tumor stages. PMID- 12582775 TI - Laparoscopic management of Meckel's diverticulum in adults. AB - BACKGROUND: Meckel's diverticulum is an uncommon entity. A high index of suspicion is necessary for opportune diagnosis and prompt treatment. Technetium (TC) 99m pertechnetate scintigraphy is a sensitive and specific test for Meckel's diverticulum. In adults, the scan contributes little to clinical decision making and often will not change the need for surgical intervention. We describe our experience with four patients. METHODS: Between August 2000 and August 2001, four patients were seen with Meckel's diverticula. Three were male and one was female. The mean age was 39 years (range, 18-64). Three patients presented with anemia and one with an acute abdomen. A 99mTc pertechnetate scan was performed at a cost of 900 dollars in the three anemic patients after other endoscopic and radiographic tests were nondiagnostic. Only one patient had a positive scan. All four patients underwent exploratory laparoscopy and small bowel resection. In one patient, a minilaparatomy had to be performed. RESULTS: All patients had a satisfactory outcome without complications. Three patients were discharged within 3 days of surgery. The remaining patient had a prolonged hospital stay because of ongoing chemotherapy for small cell lung cancer. In the three anemic patients who underwent enterectomy, ulcerated small bowel outside the diverticulum was found by the pathologist. CONCLUSION: Laparoscopy is safe, cost-effective, and efficient for the diagnosis and definitive management of Meckel's diverticulum. Technetium 99m pertechnetate scintigraphy scanning adds considerable time and expense to the care of the patient without significant benefits in adults. The practice of exploratory laparoscopy rather than scintigraphy is recommended. PMID- 12582776 TI - Current status of an antireflux procedure in laparoscopic Heller myotomy. AB - BACKGROUND: Persistent dysphagia and postoperative gastroesophageal reflux (GER) are the most cited reasons for surgical failure of laparoscopic Heller myotomy. Adding an antireflux procedure to Heller myotomy has been proposed to prevent reflux. We hypothesized that an antireflux procedure added to laparoscopic Heller myotomy has little effect on preventing the symptoms or long-term sequelae of GER in achalasia patients. METHODS: We performed a meta-analysis of studies on human subjects reported in the English language literature from 1991 to 2001 years. RESULTS: An antireflux procedure accompanied laparoscopic myotomy in 15 studies with 532 patients. In 6 studies of 69 patients, no antireflux procedure was added to laparoscopic myotomy. Follow-up was available on 489 patients (92%) with partial fundoplication. The rate of GER diagnosed in pH studies was 7.9% (18 of 228 patients studied), whereas only 5.9% of patients experienced symptoms of GER (29 of 489 patients followed). Of the 69 patients without fundoplication, 47 (68%) were available for follow-up. Forty patients (85%) were studied with pH monitoring postoperatively, with 4 (10%) demonstrating reflux. Six (13%) of 47 patients had symptoms of GER. The difference in the rate of GER diagnosed in postmyotomy pH studies in wrapped and nonwrapped patients was not significant (7.9 vs 10%, respectively; p = 0.75). There was also no significant difference in the incidence of postmyotomy GER symptoms in wrapped and nonwrapped patients (5.9 vs 13% respectively; p = 0.12). CONCLUSIONS: Reflux is not necessarily eliminated with the addition of a partial fundoplication. Based on the published data, recommendations cannot be made regarding the efficacy of adding an antireflux procedure to laparoscopic Heller myotomy. Prospective randomized study is needed to clarify the role of an antireflux procedure after laparoscopic Heller myotomy. PMID- 12582777 TI - Gallstone in a hernia sac. AB - Perforation of the gallbladder with spillage of stones into the abdominal cavity is relatively common during laparoscopic cholecystectomy. We report a patient presenting with a symptomatic groin hernia 10 days after laparoscopic cholecystectomy for acute cholecystitis. The hernia sac was found to contain a gallstone. The patient underwent an uneventful hernia repair. Various complications due to retained gallstones have been described as case reports emphasizing how important it is to prevent perforation of the gallbladder or else make an effort to retrieve spilled stones from the abdomen. PMID- 12582778 TI - Endoscopic obliteration of a recurrent tracheoesophageal fistula with enbucrilate and polidocanol in a child. AB - We present a difficult case of a recurrent tracheoesophageal fistula following primary surgical repair of esophageal atresia. After four unsuccessful attempts to close the fistula, which included three thoracotomies and one endoscopic obliteration using fibrin glue, successful recurrent tracheoesophageal fistula closure was attained with the endoscopic application of enbucrilate (Histoacrylate) combined with polidocanol. Enbucrilate was applied into the lumen of the fistula by bronchoscopy, and polidocanol (Sclerovein) was injected into the esophageal submucosa around the fistula by esophagoscopy. Three years after this treatment, the fistula remains apparently closed. PMID- 12582779 TI - Comparison of endoscopic glottis-dilating operations. AB - Endoscopic glottis-dilating operations were first utilized in 1948 by Thornell, who performed an endolaryngeal arytenoidectomy. The real breakthrough in these operations was reported by Kleinsasser in 1968. There have been many modifications of the endoscopic glottis dilating operations by other authors over the past 30 years or more. These methods have brought great progress in relieving airway obstruction. However, some disadvantages have reduced the effectiveness of these operations. This study will compare the advantages and disadvantages of the previous methods and compare them to the methods based on the endo-extralaryngeal suture technique by Lichtenberger in the hope that some of the previous ineffectiveness of glottis-dilating operations can be eliminated. These recent endo-extralaryngeal suture techniques consist of two operations. The first operation performed on patients whose vocal cords were paralyzed is an irreversible operation. This was performed with and without arytenoidectomy. These operations were successful in 89 out of 94 patients. The second operation was reversible endo-extralaryngeal lateralization, which was carried out in 37 patients; of these operations, 35 were successful. The operation was performed, and, if the cords remained paralyzed, the suture was not removed. If there was evidence of a return of vocal cord function, the suture was removed, eliminating the need for further dilating operations. The author feels that these two operations are quite successful, because the medial mucous membrane of the vocal cord is preserved, and this avoids the scar and granuloma formation that are characteristic of most other glottic dilating operations. PMID- 12582780 TI - Exploration of the early auditory effects of hyperlipoproteinemia and diabetes mellitus using otoacoustic emissions. AB - Although the relationship between hearing loss and hyperlipoproteinemia (HLP) or diabetes mellitus (DM) has been shown in many clinical investigations, this concept is still controversial. A prospective study was designed to search for the existence of subclinical auditory dysfunction related to HLP and DM by transient-evoked (TEOAEs) and distortion-product otoacoustic emissions (DPOAEs) in patients with hearing levels better than 30 dB. Evoked otoacoustic emissions were utilized to investigate subclinical auditory dysfunction. Fifteen hypercholesterolemic patients (28 ears), 21 hypertriglyceridemic patients (42 ears) and 21 DM patients (40 ears) were eligible for investigation. The results of the DPOAEs and TEOAEs of the study groups were compared with the control group composed of individuals with similar ages and with normal blood lipids and glycemia. This group consisted of 22 people (44 ears). There was no difference in the existence of TEOAEs at all frequencies among the groups (P>0.05). No differences were found in the amplitudes of the DPOAES between the groups except at 4 kHz (P>0.05). The difference was caused by the hypertriglyceridemia group (P=0.014) and the non-insulin-dependent diabetes mellitus (NIDDM) group (P=0.012) when compared with the control group. The mean DPOAE amplitudes of the hypertriglyceridemic and NIDDM groups at 4 kHz were higher than those of the control group. The decreased DPOAE amplitudes at 4 kHz in hypertriglyceridemic and diabetic patients without clinical findings are compatible with the sensorineural hearing loss observed with hyperviscosity and increased noise susceptibility, as was shown before in these patients. Longitudinal investigations should be performed with otoacoustic emissions to help with the early prediction of the prospective effects of HLP and DM on the auditory system. PMID- 12582781 TI - The relationship between the intraoperative ECAP threshold and postoperative behavioral levels: the difference between postlingually deafened adults and prelingually deafened pediatric cochlear implant users. AB - The purpose of this study was to determine the relationship between electrically evoked compound-action potential (ECAP) thresholds, electrically evoked auditory brain-stem response (EABR) thresholds, behavioral thresholds (T levels) and maximum comfort levels (C levels) in profoundly deaf cochlear-implant users. The ECAP thresholds were measured intraoperatively in eight postlingually deafened adults and nine (eight prelingually and one postlingually deafened) children implanted with the Nucleus CI24 M cochlear implant. The mean ECAP thresholds did not differ between children and adults. The average behavioral T and C levels after at least 6 months of experience with a cochlear implant were significantly higher in children than those in adults. The ECAP thresholds were more strongly correlated with T and C levels in children than in adults. The stronger correlation between ECAP thresholds and behavioral T and C levels in children than in adults might result from differences in loudness sensation, which should in turn depend on auditory experience. PMID- 12582782 TI - Betahistine dihydrochloride in the treatment of peripheral vestibular vertigo. AB - The present study compares the efficacy and safety of betahistine dihydrochloride to that of a placebo in recurrent vertigo resulting from Meniere's disease (MD) or in paroxysmal positional vertigo (PPV) of probable vascular origin. The design was double-blind, multicentre and parallel-group randomised. Eleven Italian centres enrolled 144 patients: 75 of the patients were treated with betahistine (41 MD/34 PPV) and 69 with placebos (40 MD/29 PPV). The betahistine dosage was 16 mg twice per day for 3 months. Compared to the placebo, betahistine had a significant effect on the frequency, intensity and duration of vertigo attacks. Associated symptoms and the quality of life also were significantly improved by betahistine. Both the physician's judgement and the patient's opinion on the efficacy and acceptability of the treatment were in agreement as to the superiority of betahistine. The effective and safe profile of betahistine in the treatment of vertigo due to peripheral vestibular disorders was confirmed. PMID- 12582783 TI - Screening of middle ear effusion for the common sinus pathogen Bipolaris. AB - Recently, a third of middle ear effusions have been shown to harbor fungal DNA by polymerase chain reaction (PCR). This suggests that fungi, in addition to being an important sinus pathogen, may also play an important role in acute and serous otitis media. Bipolaris is an important sinus pathogen whose role in infections of the ear is unknown. In this study, we assessed if Bipolaris DNA was present in 19 middle ear effusions that were PCR positive for the presence of fungi. Primer pair specific for Bipolaris spicifera was tested against DNA from various bacterial and fungal species to demonstrate its specificity and was subsequently used on DNA isolated from effusions to determine if Bipolaris-specific DNA was present. None of the nineteen specimens tested positive for Bipolaris by PCR or standard culture technique. This suggests that while fungi may play an important role in otitis media, this study does not support a role for Bipolaris as a middle ear pathogen and may reflect regional differences in its prevalence. PMID- 12582784 TI - A patient database application for Hereditary Deafness Epidemiology and Clinical Research (H.E.A.R.): an effort for standardization in multiple languages. AB - One of the most challenging and neglected issues in medicine is the effective recording of the data obtained from the patients. The "European Work Group on the Genetics of Hearing Impairment," which has been working since 1996, proposed a few questionnaires to collect data regarding the phenotype, ENT findings, audiological examination findings and other special investigations. In this study, a computerized patient database application named "Izmir H.E.A.R version 1.0," written in Delphi 4.0 for Windows for recording the patients with hearing problems, is presented. The application consists of a modular form, including information about identity, genetic condition, proband query, audiology and vestibular tests, phenotype, pedigree and special examinations, which allows data entry on all these issues. It has been developed by using the guidelines of Hereditary Deafness Epidemiology and Clinical Research (H.E.A.R.) and by the experience gained within the last 10 years by the authors. The target population of the program is the ENT clinicians, audiologists, epidemiologists, geneticists and researchers in the field. The main idea is to create a program serving the needs of both the daily routine work and research purposes and to distribute this program to the above-mentioned specialists, to encourage them to try the first version and to find a standard and/or better way to collect data. For this reason, the program aims to be multilingual, and the currently available languages are English, German, Spanish and Turkish. PMID- 12582785 TI - Herpes zoster-associated trigeminal trophic syndrome: a case report and review. AB - The case of a 75-year-old Caucasian woman with a trigeminal trophic syndrome (TTS) is presented and discussed. TTS of the ala nasi subsequent to a herpes zoster infection has not been described previously. We provide a review of the literature with insights into the pathogenesis and current therapeutic strategies. PMID- 12582786 TI - Results of uvulopalatopharyngoplasty after diagnostic workup with polysomnography and sleep endoscopy: a report of 136 snoring patients. AB - We subjectively and objectively evaluated 136 patients with socially unacceptable snoring (SUS) or obstructive sleep apnoea syndrome (OSAS) treated with uvulopalatopharyngoplasty (UPPP) after a diagnostic workup by sleep registration (polysomnography, PSG) and sleep endoscopy. Of the 136 patients, there were 88 with OSAS and 48 with SUS. The results of the procedure were considered subjectively to be an improvement in 38 (79%) of the SUS patients and in 74 (84%) of the patients with OSAS. In 36 (40%) of the 88 patients with OSAS, repeating PSG postoperatively was considered unnecessary because of obvious improvement. Of the 52 patients with a measurement after UPPP, a decrease in the apnoea hypopnoea index (AHI) was found in 38 (73%; median decrease: 48%), and AHI dropped below 20 in 32 (62%). The apnoea index (AI) was available in 49 (56%) patients and was reduced in 31 (63%; median decrease: 73%). An overall positive result in the 88 patients with OSAS (combining available data on subjective and objective results) was therefore found in 61 (69%; positive subjective result and AHI <15) or 71 (81%; positive subjective result and decrease in AHI), respectively, depending on the definition. We conclude that after diagnostic workup by sleep registration and sleep endoscopy, the success rate of UPPP increases as compared to historical controls. PMID- 12582787 TI - Malignant pleomorphic adenoma (malignant mixed tumor) of the trachea: a case report and review of the literature. AB - A case of malignant mixed tumor of the trachea in a 56-year-old man is described. His tumor was removed by segmental tracheal resection, and end-to-end anastomosis was performed. Histologically, the tumor was characterized by a biphasic composition showing admixtures of epithelial elements in varying proportions; these were cytologically atypical with prominent mitotic figures. However, stromal elements were osteoid and mixoid with a benign appearance. The patient had no evidence of disease in the head and neck region and had no history of previous surgery for a salivary gland tumor. These findings were interpreted as indicative of malignant pleomorphic adenoma of the trachea. PMID- 12582788 TI - Influence of stability on range of motion after cruciate-retaining TKA. AB - BACKGROUND: A loosely balanced total knee arthroplasty (TKA) is reported to produce a good postoperative range of motion (ROM), but too much laxity is thought to be the cause of persistent pain and worsened functionality. METHODS: The anteroposterior and mediolateral laxity values were measured to evaluate the influence of stability after cruciate-retaining (CR) TKA on ROM and the modified Knee Society score at 4-8 years after the operation. Twenty-one knees in 15 patients with an average age of 68 years who had undergone a CR TKA for osteoarthrosis were examined. The mean preoperative and postoperative ROM was 124 degrees and 112 degrees, respectively. The mean anteroposterior and mediolateral laxity values were 9.7 mm and 10.6 degrees, respectively. RESULTS: No correlation was found between the postoperative ROM and laxity or between the modified Knee Society score and laxity. A loosely balanced TKA did not produce a good postoperative ROM. No parameters suggested that lax knees showed a higher pain score and lower functional score than stable knees. PMID- 12582789 TI - Zweymueller with metal-on-metal articulation: clinical, radiological and histological analysis of short-term results. AB - BACKGROUND: This is a prospective study. METHOD: . A total of 266 consecutive patients, who received 350 third-generation Zweymueller-SL total hip arthroplasties with metal-on-metal articulation for primary or secondary osteoarthritis, was followed and evaluated clinically, radiologically and histologically. The age of the patients at the time of surgery was 55+/-9 years, (range 25-70 years). Seven ( 3%) patients did not return for their last follow-up evaluation for reasons unrelated to their hip operation. RESULTS: The mean follow up was 52 months (range 37-92 months). The preoperative Harris hip score was 45+/ 19 and increased to 96+/-4 postoperatively. The invalidity of the patients was significantly improved postoperatively ( p<0.001). In all, 97% of the patients were satisfied or very satisfied with the result of the operation. There was no aseptic loosening noted in this series. Revision was done in 6 (1.8%) hips because of septic loosening ( n=5, 1.5%) or technical error ( n=1, 0.3%) during implantation. Dislocation of the prosthesis occurred in the early postoperative period in 2 (0.6%) hips. Periarticular ossification was observed in 30% of the hips (5% Brooker grades III and IV), but without associated disability. During revision surgery, no macroscopic metalosis could be identified in the newly formed hip joint membrane; however microscopic evidence for metalosis (Mirra grades 1 and 2) was seen in all revised hips. The survival for Zweymueller screw socket and stem 7.6 years after implantation was 99.4% and 96.8%, respectively. CONCLUSIONS: This study showed that the short-term results of Zweymueller total hip arthroplasty with metal-on-metal articulation were just as satisfactory as those of a conventional polyethylene on ceramic articulation, while the metal-on metal articulation does not seem to give rise to new problems or complications. PMID- 12582790 TI - Long-term outcome following high tibial osteotomy with tension bend principle. AB - BACKGROUND: High tibial osteotomy has been successfully performed on patients with varus gonarthrosis for many years now. The prospective study presented here serves to verify the long-term outcome of this procedure. METHODS: A supratuberous high tibial osteotomy was carried out on 100 patients with varus gonarthrosis. Since then, 5 patients have died, and 6 patients could not be traced. The regaining rate was 94%. From a total of 89 patients with 101 operated knee joints, 46 were men and 43 women. The mean age at the time of surgery was 58 years. The patients were clinically and radiologically examined 10 years postoperatively and the knee score and function score ascertained according to the International Knee Society. RESULTS: The knee score showed a statistically significant increase from 31 points prior to surgery to 79 points at the time of follow-up; the function score likewise increased from 46 points to 78 points. In addition, 77.5% of patients would undergo the same surgery again, while 79% of patients claimed their condition to have improved following surgery. The preoperative full-length weight-bearing radiograph demonstrated an axis shift in the varus position of on average 9.6 degrees at 10 years postoperatively, an overcompensation in the valgus position of 1 degrees on average. At the time of follow-up, 81% did not require repeat surgery of the knee. CONCLUSION: With a correct diagnosis and accurate surgical techniques, positive clinical and radiological long-term results can be expected. Therefore, high tibial osteotomy may constitute an important contribution to our algorithm in the treatment of varus gonarthrosis. PMID- 12582791 TI - Two-year results after lumbar microdiscectomy with and without prophylaxis of a peridural fibrosis using Adcon-L. AB - BACKGROUND: A retrospective study investigated the advantages of the intraoperative application of the anti-adhesion barrier gel Adcon-L after lumbar microdiscectomy. METHODS: For the study, 92 patients were recruited. Following the operation, 46 patients (study group) received Adcon-L application. In other 46 patients (control group), wounds were closed without Adcon-L application. Median follow-up was 32 (range 26-33) months in the study group and 37 (range 32 41) months in the control group postoperation. All patients were examined clinically, and spinal MRIs were carried out on 12 randomly selected patients in each group. RESULTS: According to the score of the Low Back Pain Committee of the Japanese Orthopedic Association, no significant clinical difference could be established between the two groups. Subjective evaluation of the operation results (score according to Weber) was not significant. Peridural scarring on the postoperative MRI of 24 patients did not differ significantly between the groups. CONCLUSION: A significant effect on the clinical results after Adcon-L application could not be found. PMID- 12582792 TI - Injectable calcium phosphate bone cement Norian SRS for the treatment of intra articular compression fractures of the distal radius in osteoporotic women. AB - BACKGROUND: Distal radius fracture often presents a metaphyseal void which is more extended in elderly, osteoporotic patients. Bone graft and bone substitutes are reported to be beneficial in maintaining metaphyseal reduction. METHODS: We performed a prospective study on 52 menopausal, osteoporotic women with unstable intra-articular distal radius fractures to compare the outcome of percutaneous pinning and immobilisation in a cast for 6 weeks with that using injectable calcium phosphate bone cement (Norian Skeletal Repair System, SRS) to supplement pin and screw fixation and immobilisation in a cast for 3 weeks. All patients were reviewed 2 years (range 21-29 months) after surgery. RESULTS: Patients treated with SRS had better functional outcome, restoration of movement and grip strength ( p<0.001). In this group there was 1 mm loss of radial length, 3 degrees loss of radial inclination and 7 degrees loss of palmar tilt. In the control group radial length decreased 3 mm, radial inclination decreased 11 degrees and palmar tilt 12 degrees. Loss of reduction was significantly higher in the control group ( p<0.001). CONCLUSION: We conclude that the use of Norian SRS to supplement pin and screw fixation is effective in maintaining the reduction of unstable intra-articular distal radius fractures in osteoporotic patients and provides a better clinical outcome than percutaneous pinning. PMID- 12582793 TI - Biological fixation of hydroxyapatite-coated versus grit-blasted titanium hip stems: a canine study. AB - BACKGROUND: The purpose of the study was to evaluate the influence of a proximal hydroxyapatite (HA) coating in comparison with a grit-blasted titanium surface of an anatomic hip stem in an animal model over a maximum duration of 2 years. METHODS: Thirty adult dogs underwent implantation of either a proximally HA coated or a grit-blasted anatomic titanium stem. The animals were clinically evaluated for their walking ability, and serial radiographs were taken. The femora were assessed histomorphologically at set time points from 6 weeks to 2 years postoperatively. Undecalcified thin section specimens through the proximal and distal portion of the coating or grit blasting were prepared. The percentage of implant surface with direct bone contact without connective tissue involvement was determined. RESULTS: Radiographically, animals with uncoated prostheses showed characteristic signs of loosening more frequently. Histomorphometrically, an average of 65% of the surface of HA-coated implants had bone contact, but only 14.7% of the surface of grit-blasted prostheses ( p=0.0001). There was no relationship between bone contact and the duration of implantation of the prosthesis, either for the coated or for the uncoated prostheses. HA coating enhances osseointegration of an anatomic hip stem. CONCLUSION: Anatomic stems with rounded design require a surface coating or surface structure, since the mere grit-blasting of the titanium surface does not ensure osseointegration in this animal model. PMID- 12582794 TI - Bilateral obturator dislocation of the hip. AB - A case of a traumatic bilateral obturator dislocation of the hip and right side acetabular fracture in a 23-year-old man is presented. The patient was managed conservatively. After follow-up time of 24 months, the result was excellent. The importance of a pelvic radiograph and physical examination of the extremities after significant major trauma are emphasized. PMID- 12582795 TI - A report of two cases treated with Pauwels' femoral osteotomy for advanced osteoarthritis resulting from a sequela of infectious coxitis in childhood. AB - Pauwels' femoral valgus osteotomy was performed on two osteoarthritic hips that developed as a sequela of infectious coxitis in the childhood. One of the hips was diagnosed as osteomyelitis of the ipsilateral femur before the operation and was treated with curettage and implantation of antibiotic-impregnated cement beads. The results of both hips were excellent and maintained for 9-11 years. PMID- 12582796 TI - Chondrosarcoma of the hand secondary to multiple enchondromatosis; report of two cases. AB - Although malignant transformation to chondrosarcoma may occur in some patients with multiple enchondromatosis, this event rarely occurs in the hand. We encountered two patients with chondrosarcoma of the hand secondary to multiple enchondromatosis. One patient was a 27-year-old man and the other, a 76-year-old man. Both patients manifested multiple osteolytic lesions in the hand on the plain radiographs. Severe bone destruction associated with a large soft-tissue swelling of the proximal and middle phalanges of the little finger was seen in case 1. In case 2, tremendous expansion and bone destruction of the middle phalanx of the ring finger was seen. Magnetic resonance images of the tumour in both patients showed low signal intensity on T1-weighted and high signal intensity on T2-weighted images. Amputation was performed in each patient. Histological examination revealed that the tumour was a grade 2 chondrosarcoma in case 1 and a grade 1 chondrosarcoma in case 2 accompanied by enchondromata. From these findings, the diagnosis of chondrosarcoma secondary to multiple enchondromatosis was made. Because quite a few patients with multiple enchondromatosis develop secondary chondrosarcoma, although rarely in the hand, the enchondromata should be curetted, unless impractical, before malignant transformation occurs. PMID- 12582797 TI - Massive metallosis after revision of a fractured ceramic head onto a metal head. AB - We report a case in which metallosis occurred due to a ceramic head fracture after revision of a total hip prosthesis. In the primary revision, after retrieval of the head fragments, the firmly incorporated stem was left in place, the PE inlay was replaced, and a metal head was inserted. Subsequently, massive metallosis occurred, leading to loosening of the stem and necessitating a two stage revision. Examinations of the explanted material showed that microparticles of the fractured ceramic head had come to rest in the new PE inlay, where they led to wear on the metal head. We conclude that the use of a metal/PE articular pairing is contraindicated after a ceramic head fracture and recommend the use of a ceramic/ceramic articular pairing in the event of revision. In the case of the stem remaining in situ, this procedure requires the use of a conical stem adapter in order to minimise the fracture risk of the new head. PMID- 12582798 TI - Morbus Teutschlander - a massive soft-tissue calcification of the foot in a patient on long-term hemodialysis. AB - Morbus Teutschlander is a benign metabolic disorder involving soft-tissue masses near large joints and a rarely observed form of extraskeletal calcifications. For example, it is seen in patients with chronic renal failure, but in most cases it remains clinically silent. In a minority of patients, this soft-tissue calcification is responsible for complications. Various locations of calcium deposits have been characterized. Advanced age and increased calcium x phosphate product are some of the underlying reasons, but local factors are involved as well. Prevention should be preferred because the treatment is generally difficult. We describe the case of a morbus Teutschlander in a 58-year-old woman with chronic renal failure on hemodialysis presenting with a massive soft-tissue calcification of the foot with a compression of a cutaneous nerve secondary due to uremic tumoral calcinosis. The tumoral masses was successfully excised, and at the 1-year follow-up, the patient had no discomfort. Clinical and radiological features and treatment are discussed, and a review of soft-tissue calcifications is given. PMID- 12582799 TI - 47th Annual Meeting of the GTH (Gesellschaft fur Thrombose- und Hamostaseforschung) 15-18 February 2003, Innsbruck, Austria. Abstracts. PMID- 12582800 TI - Osteosynthesis of distal radial fractures with a volar locking screw plate system. AB - We developed a locking screw plate system for the stabilisation of distal radial fractures, which can be inserted through a standard volar approach and in which the locking mechanism allows early post-operative mobilisation. Forty-nine patients with 50 fractures underwent surgical treatment; 66% were type C fractures. The mean follow-up was 26 months. According to the scores of Gartland and Werley and Green and O'Brien, 92% and 68% respectively had an excellent or good outcome; 46% were radiologically identical to the uninjured side and in 42% the reduction remained unchanged after 2 years. The most frequent complication was rupture of the flexor pollicis longus tendon, which occurred in six cases (12%) at a mean of 10 months after operation. PMID- 12582801 TI - Arthroplasty of the shoulder in rheumatoid arthritis with rotator cuff dysfunction. AB - We carried out a retrospective analysis of 17 total shoulder replacements using the reversed Delta III prosthesis in patients with rheumatoid arthritis of the glenohumeral joint complicated by rotator cuff dysfunction. Outcome was assessed using the Constant-Murley scoring system. In addition, general health status was assessed with the Short Form Health Survey and radiographical analysis of the prostheses undertaken. Mean age at the time of surgery was 64 years. Thirteen shoulders were followed up for more than 5 years (mean 87 months). Median Constant-Murley score was 59.0; median scores for general health were 33.40 and 49.36 for the physical and mental components respectively. Radiographical analysis revealed evidence of lucencies about the humeral component in all cases and about the glenoid component in five cases. Despite the good clinical results, the high incidence of radiographical lucencies is of concern. PMID- 12582802 TI - Nonoperative treatment of hip fractures. AB - We retrospectively reviewed a population database and a case series to compare the mortality of operative and nonoperative treatment of hip fractures in patients with severe comorbidity. Nonoperative treatment of hip fractures (bed rest or early weight bearing) was administered based on medical assessment of perioperative risk. Comparison of 30-day mortality was performed between the nonoperatively and operatively treated groups. We found that of 50,235 of hip fractures that occurred between 1992 and 1998, 89.4% were treated operatively. Thirty-day mortality rate in the nonoperatively treated patients (18.8%) was higher than the rate in operatively treated patients (11.0%) (odds ratio 1.7 times, 95% confidence interval (CI) 1.6, 1.8). In the case series, of 62 elderly patients with severe comorbidity treated nonoperatively, 41 had bed rest/traction, while 21 were mobilized early. A group of operatively treated patients (n=108) was compared to nonoperatively treated patients. Mortality with nonoperative treatment was higher with bed rest (73%) compared to early mobilization (odds ratio 3.8, 95% CI 1.1-14.0). There was no significant difference in mortality between operatively treated patients (29%) and patients treated nonoperatively with immediate mobilization (19%). Bed rest was 2.5 times more likely to be associated with mortality compared to operative treatment (95% CI 1.1-5.5). PMID- 12582804 TI - Biomechanics of anterior cruciate ligament reconstruction using twisted doubled hamstring tendons. AB - We studied the biomechanical properties of a twisted doubled semitendinosus and gracilis graft. We applied an un-axial load in order to reproduce the kinematics of a reconstructed anterior cruciate ligament (ACL). A modified cryo-jaw clamp system was used to minimize soft tissue slippage. The lower grip, after fixation of the free ends of the tendons, was rotated 45 degrees, translated 1 cm, and bent 40 degrees, simulating a knee sprain. The graft was tested to failure using a servohydraulic machine. The specimen from one knee of seven unembalmed cadavers was assigned to the untwisted (parallel) bundles group, while its pair was assigned to the twisted group. For the parallel bundles group, the mean maximum load was 1,709.3+/-581.9 N, for the twisted group 2,428.3+/-475.4 N (P<0.05). The mean stiffness was respectively 213.6+/-72.4 N/mm and 310.3+/-97.3 N/mm (P=0.08). Although caution should be used in extrapolating the results to clinical estimates of the strength of hamstring grafts, the results of the present study could justify the use of twisted semitendinosus and gracilis bundles in ACL reconstruction. PMID- 12582803 TI - Patellar component stability improves with pulsatile lavage in total knee arthroplasty. AB - In a cadaver study, we prepared 37 fresh frozen human patella pairs to determine the efficacy of jet lavage in comparison to syringe lavage with respect to cement penetration and stability of the polyethylene patellar component after patellar resurfacing. For 12 patella pairs, sagittal sections were obtained at predefined levels using a diamond saw. Cement penetration was significantly greater (P<0.0001) in the jet lavage specimens compared to the syringe lavage specimens. For the remaining 25 patella pairs, pullout tests were performed on patellar components using a traction-compression device. The maximum force required to cause mechanical failure was significantly greater (P<0,0001) in jet lavage specimens compared to syringe lavage specimens. Our results support the routine use of jet lavage for cleansing the patellar bed prior to cement application in cemented patellar resurfacing. PMID- 12582805 TI - Functional bracing in the treatment of delayed union and nonunion of the tibia. AB - We treated 16 delayed unions and 57 nonunions of the tibial diaphysis with a below-the-knee functional brace. In 48 cases, bracing was preceded by fibular ostectomy, and ten patients had an additional bone graft. We were able to follow 67 patients, of whom six (8.7%) failed to respond to treatment. In patients with nonunion, bony healing occurred with a median of 4 months. There was no difference in the speed of healing according to the level of the defect. Shortening of the limb following ostectomy of the fibula had a mean of 3 mm in the delayed-union group and 5 mm in the nonunion group. PMID- 12582806 TI - Fracture dislocations of Lisfranc's joint treated with closed reduction and percutaneous fixation. AB - We reviewed 42 patients (mean age 37.7+/-14.2 years) with closed fracture dislocations of Lisfranc's joint treated with percutaneous screw fixation. Mean follow-up was 58.4+/-17.3 months. The aim was to compare dislocations in which a perfect anatomical reduction had been reached with dislocations in which reduction was only near anatomical. The mean American Orthopaedic Foot and Ankle Society score for all patients was 81.0+/-13.5. There were no significant differences in outcome scores between patients with perfect anatomical reduction and patients with near anatomical reduction. However, patients with combined fracture dislocations obtained statistically better scores than patients with pure dislocations. PMID- 12582807 TI - The Viladot-Regnauld operation for hallux valgus. AB - Of 54 patients, we reviewed 26 with 28 operations for hallux valgus according to the Viladot-Regnauld technique. Follow-up was 18.7 (12-26) years. In 26 operations, the score according to Kitaoka and modified by Calder was excellent or good, and in two, fair. There were no complications. Average valgus angle correction was 15 degrees (8 degrees-29 degrees) and average intermetatarsal angle correction was 7 degrees (4 degrees-12 degrees). Sesamoids were medialised, on average, by at least 25% compared to the pre-operative situation. The results confirm the validity of this surgical procedure. PMID- 12582808 TI - The use of detergent irrigation for musculoskeletal wounds. AB - The primary purpose of irrigation is to remove bacterial contaminants from the wound. Surfactants do that by disrupting the bonds of the organism to the surface. The use of this wound care strategy was studied in a series of investigations spanning several years. In vitro experiments revealed that surfactant irrigation was superior to saline or antibiotic solutions for removal of adherent bacteria from metallic surfaces, from bone, and from bovine muscle. An in vivo model of the complex orthopedic wound was developed. The superiority of surfactant irrigation over saline or antibiotic solution was demonstrated in animal wounds containing metal, bone injury, and soft tissue damage. Specificity of different surfactant irrigations for various bacterial species was demonstrated. A sequential surfactant irrigation protocol was developed and shown effective in the polymicrobial wound with established infection. PMID- 12582809 TI - Distraction osteogenesis by Ilizarov and unilateral external fixators in a canine model. AB - We studied distraction osteogenesis in canine experimental model using two types of external fixators, Ilizarov (n=6) or AO unilateral (n=9) external fixator. Distraction started 1 week after surgery (2 x 0.5 mm/day) and lasted for 3 weeks. Specimens were harvested from weeks 7 through 12. The outcome was assessed by X ray, histology, histomorphometry and microradiography. Bone regeneration as observed by X-rays was satisfactory and similar in both groups. Both endochondral ossification and intramembranous ossification were found simultaneously in both groups. In both groups, bone formation parameters were significantly higher in the area of consolidating bone. No differences in histomorphometric parameters existed between the groups. In the study period, the bone formation was enhanced and prevailed in the distraction area. This study demonstrated the utility of the canine experimental model for the study of distraction osteogenesis. PMID- 12582810 TI - In vitro release of vancomycin from vancomycin-loaded blood coated demineralised bone. AB - In vitro and in vivo studies have demonstrated the possibility that cancellous bone could be used as a carrier of antibiotics for local delivery. However, the release of antibiotics from the loaded cancellous bone is too rapid and uncertain. We hypothesised that demineralisation of cancellous bone would increase the amount of antibiotic adsorbed, and coating of the freeze-dried antibiotic-impregnated cancellous bone with bio-compatible material would prolong antibiotic release. Bovine cancellous bone blocks of equal size were demineralised using a 0.5 N HCl solution and loaded with vancomycin solution under vacuum. The loaded bone blocks were then freeze-dried. To obtain a bio compatible coating, the vancomycin-impregnated bone blocks were soaked in fresh human venous blood for 3 h. The release of impregnated antibiotic from the bone blocks was evaluated in phosphate-buffered saline and foetal bovine serum. It was found that significantly larger amounts of vancomycin were adsorbed into the demineralised bone blocks than into the un-demineralised blocks. The blood coating was found to increase the duration of vancomycin release from the blocks. With demineralisation and blood coating, the blocks eluted vancomycin higher than therapeutic concentration for a 5-week period. PMID- 12582811 TI - Efficacy of contained metaphyseal and periarticular defects treated with two different demineralized bone matrix allografts. AB - The efficacies of two different allografts, Grafton (demineralized bone matrix [DBM] in a glycerol carrier) and Orthoblast (DBM in a reverse thermal poloxamer carrier) were examined from cases involving periarticular fractures. Demographic, perioperative, and outcome data for patients with periarticular fractures who underwent a prospectively designed protocol for bone grafting were compiled, with 15 cases using Orthoblast and 13 using Grafton. A successful graft was defined as healing on the first graft attempt without complications. Healing was determined by radiographic studies and clinical evaluation. The successful graft rates of Orthoblast and Grafton were 15/15 and 9/13, respectively. PMID- 12582812 TI - Treatment of painful peripheral neuromas by vein implantation. AB - Firstly, we designed a vein-implantation model using the rat femoral nerve and vein to study the morphometric changes in nerve endings inserted into venous lumina. By 4 weeks, nerve fibers had extended from the nerve stump into the lumen of the vein and along the endothelium of the vein. After 8 weeks, the lengths and number of nerve fibres extending into the vein lumen began to decrease. At 12 weeks, the nerve ending had developed a hemispherical shape. In none of the experiments was a neuroma formed. Secondly, we treated ten neuromas in ten patients by the vein-implantation method. We obtained excellent results in seven patients. PMID- 12582813 TI - Dual-modality PET/CT imaging: the effect of respiratory motion on combined image quality in clinical oncology. AB - To reduce potential mis-registration from differences in the breathing pattern between two complementary PET and CT data sets, patients are generally allowed to breathe quietly during a dual-modality scan using a combined PET/CT tomograph. Frequently, however, local mis-registration between the CT and the PET is observed. We have evaluated the appearance, magnitude, and frequency of respiration-induced artefacts in CT images of dual-modality PET/CT studies of 62 patients. Combined PET/CT scans during normal respiration were acquired in 43 subjects using single- or dual-slice CT. Nineteen patients were scanned with a special breathing protocol (limited breath-hold technique) on a single-slice PET/CT tomograph. All subjects were injected with approximately 370 MBq of FDG, and PET/CT scanning commenced 1 h post injection. The CT images were reconstructed and, after appropriate scaling, used for on-line attenuation correction of the PET emission data. We found that respiration artefacts can occur in the majority of cases if no respiration protocol is used. When applying the limited breath-hold technique, the frequency of severe artefacts in the area of the diaphragm was reduced by half, and the spatial extent of respiration induced artefacts was reduced by at least 40% compared with the acquisition protocols without any breathing instructions. In conclusion, special breathing protocols are effective and should be used for CT scans as part of combined imaging protocols using a dual-modality PET/CT tomograph. The results of this study can also be applied to multi-slice CT to potentially reduce further breathing artefacts in PET/CT imaging and to improve overall image quality. PMID- 12582814 TI - Influence of fast lymphatic drainage on metastatic spread in cutaneous malignant melanoma: a prospective feasibility study. AB - The concept of sentinel lymph node biopsy in cutaneous malignant melanoma is widely established. Preoperative cutaneous lymphoscintigraphic mapping is a reliable method for identifying the nodal basins at risk of metastases in melanomas. In this prospective study we investigated the correlation between the scintigraphic appearance time and the metastatic involvement of sentinel lymph nodes. In 276 malignant melanoma patients (137 women, 139 men; age 16-93 years), dynamic and static lymphoscintigraphy was performed after strict intracutaneous application of technetium-99m nanocolloid (40-150 MBq; 0.05 ml/deposit) around the tumour or biopsy scar. Analysis of dynamic scans primarily focussed on the appearance time of sentinel lymph nodes. Sentinel lymph node visualisation 20 min as slow drainage. Fast lymphatic drainage was found in 236 patients, of whom 34 (14.4%) had sentinel lymph node metastases. Twenty-two patients showed hybrid (fast and slow) lymphatic drainage, and eight (36.4%) of them had sentinel lymph node metastases. Seven of the latter demonstrated fast lymphatic drainage, while one showed one positive sentinel lymph node with fast and another with slow drainage. The melanomas of 18 patients demonstrated exclusively slow lymphatic drainage, in all cases without sentinel lymph node metastases. This prospective study indicates that the scintigraphic appearance time of sentinel lymph nodes seems to be a clinically relevant factor for prediction of metastatic spread of cutaneous malignant melanoma. Larger numbers of patients need to be examined to truly assess the benefit of the scintigraphic appearance time compared with other predictors of sentinel lymph node tumour positivity. PMID- 12582815 TI - 86Y-DOTA0)-D-Phe1-Tyr3-octreotide (SMT487)--a phase 1 clinical study: pharmacokinetics, biodistribution and renal protective effect of different regimens of amino acid co-infusion. AB - The pharmacokinetics and dosimetry of (86)Y-DOTA(0)- d-Phe(1)-Tyr(3)-octreotide ((86)Y-SMT487) were evaluated in a phase I positron emission tomography (PET) study of 24 patients with somatostatin receptor-positive neuroendocrine tumours. The effect of amino acid (AA) co-infusion on renal and tumour uptake was assessed in a cross-over randomised setting. Five regimens were tested: no infusion, 4-h infusion of 120 g mixed AA (26.4 g l-lysine + l-arginine), 4 h l-lysine (50 g), 10 h 240 g mixed AA (52.8 g l-lysine + l-arginine) and 4 h Lys-Arg (25 g each). Comparisons were performed on an intra-patient basis. Infusions of AA started 0.5 h prior to injection of (86)Y-SMT487 and PET scans were obtained at 4, 24 and 48 h p.i. Absorbed doses to tissues were computed using the MIRD3 method. (86)Y SMT487 displayed rapid plasma clearance and exclusive renal excretion; uptake was noted in kidneys, tumours, spleen and, to a lesser extent, liver. The 4-h mixed AA co-infusion significantly ( P<0.05) reduced (86)Y-SMT487 renal uptake by a mean of 21%. This protective effect was significant on the dosimetry data (3.3+/ 1.3 vs 4.4+/-1.0 mGy/MBq; P<0.05) and was further enhanced upon prolonging the infusion to 10 h (2.1+/-0.4 vs 1.7+/-0.2 mGy/MBq; P<0.05). Infusion of Lys-Arg but not of l-lysine was more effective in reducing renal uptake than mixed AA. Infusion of AA did not result in reduced tumour uptake. The amount of (90)Y SMT487 (maximum allowed dose: MAD) that would result in a 23-Gy cut-off dose to kidneys was calculated for each study: MAD was higher with mixed AA co-infusion by a mean of 46% (10-114%, P<0.05 vs no infusion). In comparison with 4 h mixed AA, the MAD was higher by a mean of 23% (9-37%; P<0.05) with prolonged infusion and by a mean of 16% (2-28%; P<0.05) with Lys-Arg. We conclude that infusion of large amounts of AA reduces renal exposure during peptide-based radiotherapy and allows higher absorbed doses to tumours. The prolongation of the infusion from 4 to 10 h further enhances the protective effect on the kidneys. PMID- 12582816 TI - Simulations of NMR-detected diffusion in suspensions of red cells: the "signatures" in q-space plots of various lattice arrangements. AB - Coherence effects from pulsed field-gradient spin-echo (PGSE) nuclear magnetic resonance diffusion experiments have been observed and characterized for diffusants in many heterogeneous systems, ranging from porous materials to cell suspensions. The resulting coherence patterns appear in plots of the normalized PGSE signal intensities as a function of the spatial wave vector Q in a so-called q-space plot. The origin of these phenomena and their mathematical and physical underpinnings are now well established. We have conducted a number of studies of diffusion-coherence phenomena in suspensions of red blood cells and have made extensive use of computer simulations of molecular diffusion in virtual lattices of cells to aid in the interpretation and analysis of experimental data. In the current work we extended the canonical model used in these studies to investigate the effect that varying the packing arrangement of cells in the suspension has on the coherence patterns, as seen in q-space plots. We show that changes in the packing arrangement of cells are reflected in the q-space plots and in the results of diffusion tensor analysis and thus we speculate upon the possible clinical importance of these findings. PMID- 12582817 TI - Structural, dynamic and mechanical properties of POPC at low cholesterol concentration studied in pressure/temperature space. AB - We have studied the structural, dynamic and mechanical properties of 1-palmitoyl 2-oleoyl- sn-glycero-3-phosphatidylcholine (POPC)/cholesterol binary mixtures by small-angle X-ray scattering. Our investigations were concentrated on the biologically most relevant pressure-temperature-cholesterol regime, i.e. the liquid crystalline phase and its phase boundary to the lamellar gel phase within a cholesterol concentration up to 25 mol%. From the dependence of the transition pressure we derived a value of 19 kJ/mol for the transition enthalpy Delta H(m) of POPC in excess water. With increasing cholesterol concentration, Delta H(m) drops to about 7 kJ/mol at 20 mol% cholesterol. Time-resolved pressure-scan (p scan) and temperature-jump (T-jump) experiments reveal that at low cholesterol content (<5-8 mol%) the fluidity and also the bilayer compressibility increase remarkably. In contrast, at concentrations between 5 and 25 mol% cholesterol the bilayer becomes again more rigid and the lipid bilayer spacing increases about 2 A. Theses changes are attributed to the onset of phase separation between liquid disordered and liquid ordered phases. The fluid-fluid miscibility gap for this mono-unsaturated lecithin species is strongly enlarged compared with saturated lecithins. PMID- 12582818 TI - Are there temperature-dependent structural transitions in the "intrinsically unstructured" protein prothymosin alpha? AB - Prothymosin alpha, a typical member of the class of the so-called "intrinsically unstructured" proteins, adopts a random-chain conformation under physiological environmental conditions. An apparent formation of ordered secondary structure and a moderate compaction are observed upon the change from neutral to acid pH at room temperature. We have addressed the question of whether there are temperature dependent changes of the conformational state of prothymosin alpha at low pH using circular dichroism spectroscopy and static and dynamic light scattering. In contrast to previous investigations, we did not observe a heat-induced conformational transition. For comparison, we have also carried out the same experimental procedures with acid-unfolded phosphoglycerate kinase from yeast. In this case we observed a weak compaction and a slight apparent increase in ordered secondary structure with increasing temperature, probably caused by the higher average hydrophobicity as compared to prothymosin alpha. In the absence of a clear structural transition, we deduce the observed effects result mainly from a progressive redistribution in the population of phi-psi angles of the polypeptide backbone when the temperature is increased. Furthermore, the paper should demonstrate the difficulties in distinguishing between such a progressive change amongst a continuum of states within the ensemble of unfolded conformations from the formation of authentic stable secondary structures in highly unfolded proteins. This problem is not solved presently and convincing evidence can only be supplied by the combination of various experimental techniques. PMID- 12582819 TI - A water network within a protein: temperature-dependent water ligation in H64V metmyoglobin and relaxation to deoxymyoglobin. AB - The sperm whale myoglobin mutant H64V, where the distal histidine is mutated to valine, is known to be five coordinated in the ferric state at room temperature and physiological pH. A change of the ligation in this H64V-Mbmet has been observed by optical absorption spectroscopy as a function of temperature from 20 K to 300 K. Above the dynamical transition at about 180 K one observes the temperature-dependent equilibrium between five- and six-ligated heme. Below the dynamical transition the equilibrium is frozen-in at about 50% of six-coordinate molecules. The water ligation of the iron occurs at temperatures where protein specific motions are present, as monitored by Mossbauer spectroscopy. The X-ray structures of H64V-Mbmet at 300 K and 110 K are reported with a resolution of 1.5 A and 1.3 A, respectively. The measurements at high resolutions are possible owing to crystallization in the space group P2(1), whereas all mutant myoglobins studies up to now have been carried out with crystals in the space group P6. The overall structure at both temperatures is very close to the native myoglobin. The binding of water at the sixth coordination site at lower temperatures is possible owing to a stabilizing water network extending from the protein surface to the active centre. The reduction of the H64V-Mbmet by electrons obtained by X-ray irradiation of the water-glycerol solvent at 85 K produces an intermediate low spin state of the water-ligated molecules where Fe(II) retains the six-fold coordination. Mossbauer spectroscopy shows that the relaxation of the metastable low-spin state to high-spin H64V-Mbdeoxy with dissociation of the Fe(II)-H(2)O bond starts at about 115 K and is completed at about 170 K. Differences in the dynamics properties of the native and mutant myoglobin and the connection to the dynamical transition around 180 K are discussed. PMID- 12582820 TI - Effects of pH on kinetics of binding of mRNA-cap analogs by translation initiation factor eIF4E. AB - Stopped-flow spectrofluorimetry and a theoretical method for predicting protonation equilibria in polyelectrolytes were combined in an analysis of the pH dependence of the kinetics of binding of analogues of the 5'-mRNA cap to the cap binding protein eIF4E. The computer simulations and available experimental data indicate that there are two titratable groups in the binding site of the protein and two titratable groups on the ligands directly involved in the binding, in addition to stacking interactions described by other groups. The observed pH dependencies of the rate constants obtained from the stopped-flow experiments are consistent with this finding. In particular, it is concluded that binding of both forms of the cap analogs regarding protonation at the N1 position of the guanine ring is efficient, and the shift to a predominantly protonated form of the ring takes place after formation of the complex. PMID- 12582821 TI - pH-dependent stability of sperm whale myoglobin in water-guanidine hydrochloride solutions. AB - An experimental-theoretical approach for the elucidation of protein stability is proposed. The theoretical prediction of pH-dependent protein stability is based on the macroscopic electrostatic model for calculation of the pH-dependent electrostatic free energy of proteins. As a test of the method we have considered the pH-dependent stability of sperm whale metmyoglobin. Two theoretical methods for evaluation of the electrostatic free energy and p K values are applied: the finite-difference Poisson-Boltzmann method and the semiempirical approach based on the modified Tanford-Kirkwood theory. The theoretical results for electrostatic free energy of unfolding are compared with the experimental data for guanidine hydrochloride unfolding under equilibrium conditions over a wide pH range. Using the optical parameters of the Soret absorbance to monitor conformational equilibrium and Tanford's method to estimate the resulting data, it was found that the conformational free energy of unfolding of metmyoglobin is 16.3 kcal mol(-1) at neutral pH values. The total unfolding free energies were calculated on the basis of the theoretically predicted electrostatic unfolding free energies and the experimentally measured midpoints (pH(1/2)) of acidic and alkaline denaturation transitions. Experimental data for alkaline denaturation were used for the first time in theoretical analysis of the pH-dependent unfolding of myoglobin. The present results demonstrate that the simultaneous application of appropriate theoretical and experimental methods permits a more complete analysis of the pH-dependent and pH-independent properties and stability of globular proteins. PMID- 12582822 TI - Formation of monolayers and bilayer foam films from lamellar, inverted hexagonal and cubic lipid phases. AB - This study revealed large distinctions between the lamellar and non-lamellar liquid crystalline lipid phases in their spreading at the air/water interface and propensity to form bilayer foam films. Comparative measurements were made for the lamellar L(alpha), the inverted hexagonal H(II) and the bicontinuous cubic Pn3m phases of the phospholipid dipalmitoleoylphosphatidylethanolamine (DPoPE). With regard to monolayer formation, followed as the decrease of surface tension with time, the best spreading (lowest surface tension) was observed for the L(alpha) phase, and poorest spreading (highest surface tension) was recorded for the H(II) phase. The cubic Pn3m phase of DPoPE, induced by temperature cycling, retained an intermediate position between the L(alpha) and H(II) phases. According to their ability to lower surface tension and disintegrate at the air/water interface, the three phases thus order as L(alpha)>Pn3m>H(II). Clearly expressed threshold (minimum) bulk lipid concentrations, C(t), required for formation of stable foam bilayers from these phases, were determined and their values were found to correlate well with the bulk lipid phase behaviour. The C(t) values for L(alpha) and H(II) substantially increase with the temperature. Their Arrhenius plots, ln C(t) versus 1/ T, are linear and intersect at approximately 36-37 degrees C, coinciding with the onset of the bulk L(alpha)-->H(II) phase transition, as determined by differential scanning calorimetry. However, the C(t) value for the Pn3m phase, equal to 30 micro g/mL, was found to be constant over the whole range investigated between 20 degrees C and 50 degrees C. The horizontal C(t) versus T plot for the Pn3m phase crosses the respective plot for the L(alpha) phase at the temperature bounding from below the hysteretic loop of the L(alpha)<-->H(II) transition (approximately 26 degrees C), thus providing a certain insight about the thermodynamic stability of the Pn3m phase relative to the L(alpha) phase. The established strong effect of the particular lipid phase on the formation of monolayers and stable black foam films should be of importance in various in vitro and in vivo systems, where lipid structures are in contact with interfaces and disintegrate there to different extents. PMID- 12582823 TI - The bilayer melting transition in lung surfactant bilayers: the role of cholesterol. AB - Aqueous dispersions of a porcine lung surfactant (PLS) extract with and without cholesterol supplementation were analyzed by X-ray scattering. Lamellar liquid crystalline and gel-type bilayer phases are formed, as in pure phosphatidylcholine (PC)-cholesterol systems. This PLS extract, developed for clinical applications, has a cholesterol content of less than 1% (w/w). Above the limit of swelling, the bilayer structure shows a melting (main) transition during heating at about 34 degrees C. When 13 mol% cholesterol was added to PLS, so that the cholesterol content of natural lung surfactant was reached, the X-ray scattering pattern showed pronounced changes. The main transition temperature was reduced to the range 20-25 degrees C, whereas according to earlier studies of disaturated PC-cholesterol bilayers in water the main transition remains almost constant when the amount of solubilized cholesterol is increased. Furthermore, the changes in scattering pattern at passing this transition in PLS-cholesterol samples were much smaller than at the same transition in PLS samples. These effects of cholesterol solubilization can be related to phase segregation within the bilayers, known from pure PC-cholesterol systems. One phase, solubilizing about 8 mol% cholesterol, exhibits a melting transition, whereas the other bilayer phase, with a liquid-crystalline disordered conformation, has a cholesterol content in the range 20-30 mol% and this phase shows no thermal transition. The relative amount of bilayer lipids that is transformed at the main transition in the PLS-cholesterol sample is therefore only half compared to that in PLS samples. The reduction in transition temperature in the segregated bilayer of lung surfactant lipids is probably an effect of enrichment of disaturated PC species in the phase, which is poor in cholesterol. This work indicates that cholesterol in lung surfactant regulates the crystallization behavior. PMID- 12582824 TI - Enforced unbinding of biomembranes whose mutual adhesion is mediated by a specific interaction. AB - The enforced unbinding of a biomembrane from a rigid substrate whose adhesion is mediated by a specific interaction has been studied theoretically. We argue that the unbinding takes place via motion of the adhesion rim. We account for the release of the elastic energy (stored in the membrane curvature) in the binding unbinding process and obtain the dissociation constant at the rim. We further deduce an equation of motion for the rim. The solution exhibits an initial phase describing a slow motion followed by a regime of rapid motion and finally breaking of the adhesion. We show that the unbinding force depends on the rate of force application as F(*) approximately equal (F')(beta). When small forces are applied, beta=1/2, while in the case of large forces, beta=1/3. PMID- 12582825 TI - The effect of varying impact energy on diffuse axonal injury in the rat brain: a preliminary study. AB - Diffuse axonal injury (DAI) is seen as widespread damage in the white matter of brain characterized by morphological changes to axons throughout the brain and brain stem. The current study attempted to investigate the effect of increasing impact energy on the presence and severity of DAI in corpus callosum (CC). DAI was induced in adult male Sprague-Dawley rats using an injury model adapted from Marmarou et al. in 1994. A 450-g cylindrical brass weight was dropped from three different heights (2.0 m, 1.5 m and 1.0 m) on to a metal helmet affixed to the skull of the rats. In the sham group, rats underwent a surgical procedure with no impact. After a 24-h survival period the animals were transcardially perfused. The brain was removed and the cerebral hemispheres were sectioned with a vibrotome and stained by silver impregnation technique. The CC of all the impacted rats showed DAI in the form of beaded axons, retraction balls and vacuole-like enlargements. The axonal injury was most severe in the 2-m group, while mildest in the 1-m group. In the sham group, axons appeared to be normal. This study demonstrates evidence of graded DAI depending on the impact energy. Such data is useful for mathematical modeling of axonal injury in rat brain using the same impact parameters and potential determination of injury thresholds for neural trauma. PMID- 12582826 TI - Temporal precision of interceptive action: differential effects of target size and speed. AB - The duration of movements made to intercept moving targets decreases and movement speed increases when interception requires greater temporal precision. Changes in target size and target speed can have the same effect on required temporal precision, but the response to these changes differs: changes in target speed elicit larger changes in response speed. A possible explanation is that people attempt to strike the target in a central zone that does not vary much with variation in physical target size: the "effective size" of the target is relatively constant over changes in physical size. Three experiments are reported that test this idea. Participants performed two tasks: (1). strike a moving target with a bat moved perpendicular to the path of the target; (2). press on a force transducer when the target was in a location where it could be struck by the bat. Target speed was varied and target size held constant in experiment 1. Target speed and size were co-varied in experiment 2, keeping the required temporal precision constant. Target size was varied and target speed held constant in experiment 3 to give the same temporal precision as experiment 1. Duration of hitting movements decreased and maximum movement speed increased with increases in target speed and/or temporal precision requirements in all experiments. The effects were largest in experiment 1 and smallest in experiment 3. Analysis of a measure of effective target size (standard deviation of strike locations on the target) failed to support the hypothesis that performance differences could be explained in terms of effective size rather than actual physical size. In the pressing task, participants produced greater peak forces and shorter force pulses when the temporal precision required was greater, showing that the response to increasing temporal precision generalizes to different responses. It is concluded that target size and target speed have independent effects on performance. PMID- 12582827 TI - Visuomotor transformations affect bimanual coupling. AB - Interactions between bimanual movements may occur at two different levels: at a visually based level, where movement trajectories are programmed within the visually perceived external space, and at the executional level, through crosstalk of sensorimotor signals arising during movement execution. In order to distinguish between these sources of interactions, we investigated bimanual reversal movements under different conditions of visual feedback. A visuomotor transformation dissociated movement execution from visual appearance on a computer screen. The transformation we used made movements of the same amplitude evoke different excursions, and made movements of different amplitudes entail matched excursions on the screen. The transformed conditions allowed us to study which parameters of bimanual coupling were related to the way movements were executed and which correlated with the visual movement display. We found a clear dissociation between execution-related and visually related bimanual interactions. The assimilation of movement amplitudes was completely execution related. Whenever movements of different amplitudes were generated, the shorter movement was lengthened, irrespective of how the movements appeared on the feedback screen. In contrast, temporal coordination at the point of movement reversal, as well as trial-by-trial correlations of movement amplitudes, also showed significant effects of the visuomotor transformation, suggesting that these parameters are influenced by visually perceived effects of movements. This dissociation confirms the idea of separate pathways for bimanual interactions and shows that a specific set of bimanual interactions occur at least partly within a visually based external reference frame. PMID- 12582828 TI - Event-related potentials evoked by multi-feature conflict under different attentive conditions. AB - Event-related potentials (ERPs) were recorded in subjects while they were performing a matching task. The focus of the present study was on the contributions made by ERP recording to revealing the time-course of multi conflict information processing in the brain during visual attention. Participants were instructed to discriminate whether the attributes of the first stimulus (S1) were the same as those of the second one (S2) of a pair. Stimuli were defined by three features: color, global shape, and local shape. In condition 1, all attributes of the two stimuli of the pair were constant (no conflict); in condition 2, S2 was different from S1 in all attributes (conflict). The experiment comprised three tasks. In task 1, subjects attended to the color of the stimuli and ignored the other attributes. In task 2, they were asked to attend to both color and global shape of stimuli and disregard the local shape. In task 3, all attributes were attended to. An ERP component N270 was elicited by attending to the color conflict in session one. In task 2, attending to color and global shape induced two ERP effects in the conflict condition: N270 and N400. The results suggest that the attended different conflicts of the two visual attributes of the stimuli were processed in series. However, in task 3, only one apparent negative component, N270, was observed in the conflict condition. The difference between data from task 2 and 3 showed that more than one processing model exists in the human brain for processing multiple visual attribute conflicts. PMID- 12582829 TI - Binaural weighting of pinna cues in human sound localization. AB - Human sound localization relies on binaural difference cues for sound-source azimuth and pinna-related spectral shape cues for sound elevation. Although the interaural timing and level difference cues are weighted to produce a percept of sound azimuth, much less is known about binaural mechanisms underlying elevation perception. This problem is particularly interesting for the frontal hemifield, where binaural inputs are of comparable strength. In this paper, localization experiments are described in which hearing for each ear was either normal, or spectrally disrupted by a mold fitted to the external ear. Head-fixed saccadic eye movements were used as a rapid and accurate indicator of perceived sound direction in azimuth and elevation. In the control condition (both ears free) azimuth and elevation components of saccadic responses were well described by a linear regression line for the entire measured range. For unilateral mold conditions, the azimuth response components did not differ from controls. The influence of the mold on elevation responses was largest on the ipsilateral side, and declined systematically with azimuth towards the side of the free ear. Near the midsagittal plane the elevation responses were clearly affected by the mold, suggesting a systematic binaural interaction in the neural computation of perceived elevation that straddles the midline. A quantitative comparison of responses from the unilateral mold, the bilateral mold and control condition provided evidence that the fusion process can be described by binaural weighted averaging. Two different conceptual schemes are discussed that could underlie the observed responses. PMID- 12582830 TI - Adaptation of movement endpoints to perturbations of visual feedback. AB - We investigated the extent to which humans can quickly adapt their goal-directed arm movements to perturbed feedback. We predicted that the magnitude of adaptation to a changed relationship between vision and kinesthesia would depend on the type of perturbation, being largest when the perturbation can be generalized within egocentric frames of reference. To test this prediction we asked subjects to align a real 5-cm cube so that they could feel, but not see, with a simulation that they saw via a mirror. Subjects made successive movements between target locations in a sequence of adaptation and test phases. During adaptation phases, subjects received continuous visual feedback about the position of the real cube. The feedback was either veridical or perturbed. The perturbations were consistent with either a uniform translation, a scaling or a rotation. The latter two were relative to a central position between all the targets. During test phases, subjects received no visual feedback. We compared test movement endpoints after perturbed feedback with ones after veridical feedback. We found about 40% adaptation to translation, 20% to scaling and 10% to rotation. This difference in magnitude is consistent with the ease with which the transformation can be generalized within egocentric frames of reference. Changing the task so that it required different arm postures did not change the magnitude of adaptation, so postural configuration of the arm does not appear to be critical. Nevertheless, transfer to the unexposed arm was incomplete for translations and rotations, though it was complete for scaling, suggesting that at least part of the adaptation is posture based. We conclude that the adaptation to different kinds of perturbations not only differs in extent but also involves different (egocentric) mechanisms. PMID- 12582831 TI - Contribution of 5- and 12-lipoxygenase products to mechanical hyperalgesia induced by prostaglandin E(2) and epinephrine in the rat. AB - We evaluated the role of lipoxygenase products of arachidonic acid metabolism in mechanical hyperalgesia induced by epinephrine, an agent that directly sensitizes nociceptors to produce mechanical hyperalgesia via three second messenger signaling pathways, protein kinase A (PKA), protein kinase C epsilon (PKCepsilon), and mitogen activated protein kinase (MAPK). Epinephrine hyperalgesia and that induced by a selective activator of PKCepsilon, psiepsilonRACK, were inhibited by nordihydroguaretic acid (NDGA, non-selective lipoxygenase inhibitor), baicalein (BAIC, 12-lipoxygenase inhibitor) and 5, 6 dehydroarachidonic acid (5, 6-dhAA, 5-lipoxygenase inhibitor). NDGA and 5, 6-dhAA inhibited the hyperalgesia associated with activation of the protein kinase A pathway, elicited by the direct-acting hyperalgesic agent prostaglandin E(2) or by the catalytic subunit of protein kinase A. The hyperalgesia produced by active MAPK was not blocked by any of the lipoxygenase inhibitors. Injection of 5- and 12-lipoxygenase produced hyperalgesia that was not antagonized by inhibitors of PKA, PKCepsilon or MAPK. These findings suggest that: (1). lipoxygenase products of arachidonic acid function as second messengers in the peripheral hyperalgesia induced by agents that act directly on primary afferent nociceptors (epinephrine and prostaglandin E(2)), (2). products of the 5-lipoxygenase and 12-lipoxygenase pathway are involved in this function, and (3). these lipoxygenase products contribute to hyperalgesia at or downstream of protein kinase A and PKCepsilon. PMID- 12582833 TI - Trajectories of arm pointing movements on the sagittal plane vary with both direction and speed. AB - Five subjects performed arm upward and downward movements at different speeds (movement duration ranged from 0.26 to 1.2 s). Fingertip paths, velocity profiles and muscle activation patterns of arm and forearm were computed. Inspection of the electromyograph (EMG) revealed that for relatively slow speeds (>0.7 s) and for both directions, only the flexor muscles were active, mainly the anterior deltoid, for motor (upward) and braking action (downward) respectively. However, where gravity was no longer sufficient to accelerate downward and decelerate upward movements (<0.7 s), both flexors and extensors muscles were active. Path curvature and position of maximum deviation from straightness were lower for downward than for upward movements. In addition, the position of maximum deviation from straightness became progressively higher with increase in duration for both upward and downward movements. The ratio of acceleration duration to total movement duration was greater for downward than upward directions for all the range of speeds. The ratio of maximum to mean velocity was similar for upward and downward movements but decreased with decrease in speed. The results indicate that the brain accomplishes arm movements in the vertical plane with different planning processes for movements with or against gravity. Furthermore, they provide evidence that both gravitational and inertial forces are determinant for arm trajectory generation in the vertical plan. PMID- 12582832 TI - Hemispheric specialization in the co-ordination of arm and trunk movements during pointing in patients with unilateral brain damage. AB - During pointing movements involving trunk displacement, healthy subjects perform stereotypically, selecting a strategy in which the movement is initiated with either the hand or trunk, and where the trunk continues after the end of the hand movement. In a previous study, such temporal co-ordination was not found in patients with left-hemispheric brain lesions reaching with either their dominant paretic or with their non-dominant non-paretic arm. This co-ordination deficit may be associated in part with the presence of a lesion in the dominant left hemisphere. If so, then no deficit should be observed in patients with stroke related damage in their non-dominant right hemisphere moving with their ipsilesional arm. To verify this, 21 right-hand dominant adults (7 who had had a stroke in the right hemisphere, 7 who had had a stroke in the left hemisphere and 7 healthy subjects) pointed to two targets located on a table in front of them in the ipsilateral and contralateral workspace. Pointing was done under three movement conditions: while not moving the trunk, while bending the trunk forward and while bending the trunk backwards. The experiment was repeated with the non paretic arm of patients with stroke and for the right and left arms of healthy subjects. Kinematic data were recorded (Optotrak). Results showed that, compared to healthy subjects, arm-trunk timing was disrupted in patients with stroke for some conditions. As in patients with lesions in the dominant hemisphere, arm trunk timing in those with lesions in the non-dominant hemisphere was equally more variable than movements in healthy subjects. However, patients with dominant hemisphere lesions used significantly less trunk displacement than those with non dominant hemisphere lesions to accomplish the task. The deficit in trunk displacement was not due to problems of trunk control or sitting balance since, in control experiments, all subjects were able to move the trunk the required distance, with and without the added weight of the limb. Results support the hypothesis that the temporal co-ordination of trunk and arm recruitment during pointing movements is mediated bilaterally by each hemisphere. However, the difference in the range of trunk displacement between patients with left and right brain lesions suggests that the left (dominant) hemisphere plays a greater role than the right in the control of movements involving complex co-ordination between the arm and trunk. PMID- 12582834 TI - Effect of an alpha(1)-adrenergic blocker on plasticity elicited by motor training. AB - Recovery of motor function elicited by motor training after cortical lesions in rats is enhanced by norepinephrine (neurotransmitter mediating alpha(1) adrenergic function) and downregulated by alpha(1)-adrenergic antagonists. In spite of this, alpha(1)-adrenergic antagonists are used to treat elderly patients with hypertension and prostate hyperplasia in stroke settings. The purpose of this study was to determine the effects of a single oral dose of the alpha(1) adrenergic antagonist prazosin on training-dependent plasticity in intact humans, a function thought to contribute to recovery of motor function after cortical lesions. We report that prazosin decreased the ability of motor training to elicit training-dependent plasticity relative to a drug-free condition. These data suggest caution when using alpha(1)-adrenergic blockers in rehabilitative clinical settings following brain lesions. PMID- 12582835 TI - Effect of intrathecal clonidine on group I and group II oligosynaptic excitation in paraplegics. AB - We investigated the possibility that a change in transmission in group II pathways contributes to the spasticity of patients with spinal lesions. Thirteen patients were tested by measuring the quadriceps stretch reflex (Ashworth scale), the threshold of the quadriceps H reflex, and the oligosynaptic facilitation of the quadriceps H reflex elicited by volleys to groups I and II afferents in the common peroneal nerve (CPN). All these tests were performed before and after intrathecal injection of clonidine (60 microg). Early group I CPN-induced excitations occurred in 13 patients, and late group II CPN-induced excitations in 12. Both facilitations were, on average, significantly greater than those reported for normal subjects, but these increases were not correlated with the clinically assessed spasticity. Clonidine caused a constant, prolonged and dramatic decrease in spasticity, but did not alter the threshold of the quadriceps H reflex. CPN-induced group I and group II non-monosynaptic excitations of quadriceps motoneurones were significantly decreased, although they did not return to normal values. These results provide a further indication that group II pathways gives rise to the heteronymous late CPN-induced excitation. The pathophysiological role of a change in transmission in group II pathways in spasticity is discussed. PMID- 12582836 TI - Rescue of motoneurones by MK-801 innervating partially denervated rat muscles. AB - Partial denervation of the extensor digitorum longus muscle by cutting the L4 spinal nerve in 3-day-old rats causes some of the remaining uninjured motoneurones to die. A previous study has shown that of the 12 motor units usually present in the remaining uninjured L5 spinal nerve, a significantly smaller number of motor units to extensor digitorum longus muscle is found in animals operated on at 3 days. This reduction can be caused by a greater sensitivity of neonatal motoneurones with axons in a partially denervated muscle, to excitotoxic effects of glutamate. Therefore an N-methyl- D-aspartic acid (NMDA) receptor antagonist, MK-801, was injected daily for 12 days after partial denervation at 3 days. Two months after the operation contractile properties, motor unit numbers and sizes were studied. Following MK-801 treatment, the reduction in muscle weight and force output of the partially denervated muscle was less than that in the untreated group. Moreover there were more motor units in MK-801 treated animals. After partial denervation only, 15% of the total number of motor units was present whereas when the same operation was followed by treatment with MK-801, 29% remained. The mean motor unit size in the untreated group was 69% while after treatment with MK-801 it was 152% of the control. Thus treatment with MK-801 after partial denervation of neonatal animals rescued some of the motoneurones destined to die, and allowed expansion of motor unit territory of most of the surviving motoneurones. PMID- 12582837 TI - Quinacrine abolishes increases in cytoplasmic phospholipase A2 mRNA levels in the rat hippocampus after kainate-induced neuronal injury. AB - The present investigation was carried out to study the possible effects of quinacrine in modulating cytoplasmic phospholipase A(2) (cPLA(2)) mRNA levels in rat hippocampus after kainate treatment. Injections of kainate into the right lateral ventricle resulted in significant increases in cPLA(2) mRNA levels in the hippocampus, at 3 days and 7 days after injection. The elevation in cPLA(2) mRNA levels is consistent with previous observations of increased cPLA(2) immunoreactivity in degenerating neurons and astrocytes at these times. Rats that received once daily intraperitoneal injections of quinacrine (5 mg/kg) after the intracerebroventricular kainate injections showed almost complete attenuation of increased cPLA(2) expression, at both 3 and 7 days after kainate injection. These results show that in addition to its well-known effect of inhibition of PLA(2) activity, quinacrine could also inhibit cPLA(2) expression, and further supports a role for PLA(2) in kainate-induced neuronal injury. PMID- 12582838 TI - Pharmacological suppression of plastic changes in human primary somatosensory cortex after motor learning. AB - The strict division between motor and somatosensory systems might be less distinct than previously thought. Many brain mapping studies have described changes of somatosensory cortex (S-I) after the execution of a motor task, which supports the idea of a profound interconnectedness in the sensorimotor system. Here we report experiments in which we investigated by means of somatosensory evoked potentials (SSEPs) mapping the reorganizational capacities in primary somatosensory cortex before and after a Hebbian repetitive co-contraction task of the thumb and arm. We investigated the susceptibility of S-I plasticity to the pharmacological modulation of the GABA-neurotransmitter system by application of the GABA(A) agonist lorazepam. We found that repetitive training induced stable motor learning characterized by a significant improvement of performance. The time differences between the onset of contraction of the deltoid muscle and the abductor pollicis brevis were progressively shortened. The process of motor learning was accompanied by plastic changes in the primary somatosensory cortex as indicated by a significant increase in the dipole strength and a significant shift of the median nerve dipole on the hemisphere contralateral to the exercised side. Moreover, the individual shifts of median nerve dipole location were correlated with the individual improvement in motor performance. After administration of lorazepam, motor learning was significantly suppressed. The behavioural effect was accompanied by an abolition of the N20 dipole shift and an unchanged dipole strength. The results imply that motor learning leads to a profound reorganization in S-I which is subject to pharmacological suppression with the GABA agonist lorazepam. PMID- 12582839 TI - The effects of a change in gravity on the dynamics of prehension. AB - Investigating cyclic vertical arm movements with an instrumented hand-held load in an airplane undergoing parabolic flight profiles allowed us to determine how humans modulate their grip force when the gravitational and the inertial components of the load force are varied independently. Eight subjects participated in this study; four had already experienced parabolic flights and four had not. The subjects were asked to move the load up and down continuously at three different gravitational conditions (1 g, 1.8 g, and 0 g). At 1 g, the grip force precisely anticipated the fluctuations in the load force, which was maximum at the bottom of the object trajectory and minimum at the top. When gravity changed, the temporal coupling between grip force and load force persisted for all subjects from the first parabola. At 0 g, the grip force was accurately adjusted to the two load force peaks occurring at the two opposite extremes of the trajectory due to the absence of weight. While the experienced subjects exerted a grip force appropriate to a new combination of weight and inertia since their first trial, the inexperienced subjects dramatically increased their grip when faced with either high or low force levels for the first time. Then they progressively released their grip until a continuous grip load force relationship with regard to 1 g was established after the fifth parabola. We suggest that a central representation of the new gravitational field was rapidly acquired through the incoming vestibular and somatic sensory information. PMID- 12582840 TI - Vergence responses to forward motion in monkeys: visual modulation at ultra-short latencies. AB - In two monkeys, we measured the initial vergence eye movements elicited by sudden forward motion on a linear sled. Animals faced a tangent screen and experienced the translation while in darkness, fixating a small, centered spot, or viewing a large-field pattern. Forward movements elicited convergent responses that were enhanced in the presence of the visual stimuli after a latent period. The enhancement was greater with the large-field pattern than with the small spot. The latencies of these visual effects were ultra-short and less than those reported for the vergence eye movements elicited by pure visual stimuli when applied in isolation. It is possible that these ultra-short latencies resulted from the fact that there were multiple visual cues available to sense the change in viewing distance, including binocular disparity, radial optic flow, size changes, and blur. Another possibility is that the very earliest visual effects during forward motion resulted from direct modulation of the otolith-mediated, translational vestibulo-ocular reflex. PMID- 12582841 TI - Alternating optokinetic nystagmus (OKN) induced by intermittent display of stationary gratings. AB - In this paper, we report a novel optokinetic nystagmus (OKN), which was evoked by stationary gratings presented intermittently. OKN eye movements were accurately measured by the electromagnetic scleral search-coil technique. For the luminance stimuli, alternating OKN was elicited when the interstimulus interval (ISI) ranged from 33 to 83 ms duration and the ISI luminance approximated to the mean luminance of the stimulus grating; for chromatic (red/green) stimuli, the OKN could be evoked in non-isoluminant condition and vanished in the isoluminant condition. It is a plausible explanation that the present OKN, intermittent display-of-gratings-induced OKN (IDG-OKN), might be related to the temporal impulse response of the luminance channel in vision. PMID- 12582842 TI - Effects of barley yellow mosaic disease resistant gene rym1 on the infection by strains of Barley yellow mosaic virus and Barley mild mosaic virus. AB - Although a Chinese landrace of barley, Mokusekko 3, is completely resistant to all strains of Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV), and is known to have at least two resistant genes, rym1 and rym5, only rym5 has been utilized for BaYMV resistant barley breeding in Japan. In order to clarify the effect of rym1 on BaYMV and BaMMV, and to utilize the gene for resistant barley breeding, the susceptibilities of only rym1 carrying breeding lines against BaYMV and BaMMV were investigated. In the assessment of resistance to BaYMV-I, 341 F(2) populations derived from a cross between the resistant line Y4 with only rym1 and the susceptible cv Haruna Nijo shows that the segregation loosely fits a 1R:3S ratio (0.05 > P > 0.01), suggesting that the resistance is controlled by a single recessive gene, rym1. Further, none of the F(3) lines derived from the nine resistant F(2) plants showed any disease symptoms in the field infected by BaYMV-I. The same nine F(3) lines showed almost the same agronomic characters in the field infected by BaYMV-III as those in the uninfected field, apart from the symptom of showing numerous mosaics. This result indicates that the gene rym1 has an acceptable level of resistance to BaYMV-III. In the assessment of resistance to BaYMV-II, BaMMV-Ka1 and -Na1, an artificial infection method was adopted and the susceptibilities to those viruses were investigated. Although the control varieties, Ko A and Haruna Nijo, were infected with all of them, the rym1 gene carrying BC(2)F(3) lines were completely resistant to all strains. In summary, rym1 is completely resistant to BaYMV-I, II, BaMMV-Ka1 and -Na1, and has an acceptable level of resistance to BaYMV-III. This study concludes with a discussion of the reason why the important resistance gene rym1 was eliminated along with resistant cultivars during breeding for resistance to BaYMV. PMID- 12582843 TI - ESTs as a source for sequence polymorphism discovery in sugarcane: example of the Adh genes. AB - Expressed sequence tags (ESTs) have proven to be a valuable tool to discover single nucleotide polymorphism (SNP) in human genes but their use for this purpose is still limited in higher plants. Using a database of approximately 250,000 sugarcane ESTs we have recovered 219 sequences encoding alcohol dehydrogenases ( Adh), which tagged 178 distinct cDNAs from 27 libraries, constructed from at least four different cultivars. The partitioning of these ESTs into paralogous genes revealed three Adh genes expressed in sugarcane, one Adh2 and two Adh1. The soundness of the partition was carefully checked by comparison to external data, especially from the closely related sorghum. Analysis of polymorphism in the alignments of EST sequences revealed a total of 37 highly reliable SNPs in the coding and untranslated regions of the three Adh genes. In the coding regions, the mean occurrence of SNPs was one for every 122 base pair. A total of eight insertion-deletions was observed, their occurrence being limited to untranslated regions. These results show that EST data constitute an invaluable source of sequence polymorphism for sugarcane that is worth carefully collecting for the future development of new marker tools. PMID- 12582844 TI - Chromosomal localization of 5S and 18S-5.8S-25S ribosomal DNA sites in five Asian pines using fluorescence in situ hybridization. AB - Fluorescence in situ hybridization (FISH) was employed on mitotic metaphase chromosome preparations of five Asian Pinus species: Pinus tabuliformis, Pinus yunnanensis, Pinus densata, Pinus massoniana and Pinus merkusii, using simultaneously DNA probes of the 18S rRNA gene and the 5S rRNA gene including the non-transcribed spacer sequences. The number and location of 18S rDNA sites varied markedly (5-10 pairs of strong signals) among the five pines. A maximum of 20 major 18S rDNA sites was observed in the diploid genome (2n = 24) of P. massoniana. The 5S rDNA FISH pattern was less variable, with one major site and one minor site commonly observed in each species. The differentiation of rDNA sites on chromosomes among the five pines correlates well with their phylogenic positions in Pinus as reconstructed from other molecular data. P. densata, a species of hybrid origin, resembles its parents ( P. tabuliformis and P. yunnanensis), including some components characteristic of each parent in its pattern. However, the species is unique, showing new features resulting possibly from recombination and genome reorganization. PMID- 12582845 TI - Integration of common bean ( Phaseolus vulgaris L.) linkage and chromosomal maps. AB - Fluorescent in situ hybridisation of pooled, closely linked RFLP markers was used to integrate the genetic linkage map and the mitotic chromosome map of the common bean. Pooled RFLP probes showed clear and reproducible signals and allowed the assignment of all linkage groups to the chromosomes of two Phaseolus vulgaris cultivars, Saxa and Calima. Low extension values for signals originating from clustered RFLPs suggest that these clones are physically close to each other and that clusters in the genetic map are not a result of suppression of recombination due to the occurrence of chromosome rearrangements. For linkage group K, clustering of markers could be associated with proximity to centromeres. High variation in the number of 45S rDNA loci was observed among cultivars, suggesting that these terminal sites are highly recombinogenic in common bean. PMID- 12582846 TI - Identification of chromosome arms influencing expression of the HMW glutenins in wheat. AB - The high-molecular-weight (HMW) glutenin genes, located on the group 1L chromosome arms, are a major determinant for baking quality in wheat ( Triticum aestivum L.). In addition, the HMW glutenin genes provide a valuable model system for studying the evolution and regulation of orthologous and paralogous genes in polyploid species. The goal of this study was to identify loci that modify the expression of the HMW glutenins, and to map them to specific chromosome arms. Comparisons were made between endosperms with zero versus three (or three versus six) doses for each of the 42 chromosome arms of wheat. SDS-PAGE and scanning densitometry were used to quantify the protein expression levels of the four HMW glutenin genes in cv. Chinese Spring, for each of the dosage comparisons. Fifteen chromosome arms were found to have significant effects on Glu-B1-1, excluding the structural gene dosage effect: eight positive effects on 1AL, 2AS, 2BL, 2DS, 5DS, 6AL, 6DL, and 7AL and seven negative effects on 1BS, 1DS, 1DL, 4DL, 6BS, 6DS, and 7AS. Nineteen chromosome arms had significant effects on Glu-B1-2, excluding the structural gene dosage effect: eight positive effects on 1AL, 2AS, 2BS, 3AL, 4BL, 6DS, 7BL and 7DS and 11 negative effects on 1AS, 1BS, 1DS, 1DL, 2AL, 2BL, 3DS, 4BS, 4DL, 5BL, and 6BS. Twenty chromosome arms had significant effects on Glu-D1 1, excluding the structural gene dosage effect: 11 positive effects on 1AL, 1BL, 2BS, 2DS, 5BS, 5DS, 6AL, 6DS, 6DL, 7AL, and 7BL and nine negative effects on 1AS, 1BS, 1DS, 2BL, 4DL, 5BL, 5DL, 6BL, and 7DS. Twenty-five chromosome arms had significant effects on Glu-D1-2, excluding the structural gene dosage effect: 17 positive effects on 1BL, 2AS, 2BS, 2DS, 2DL, 3AS, 3AL, 3BS, 5AS, 5BS, 5DL, 6AL, 6DL, 7AL, 7BS, 7BL, and 7DL and eight negative effects on 1DS, 4DL, 5AL, 5BL, 6BS, 6BL, 6DS and 7DS. Of the 164 gene-chromosome arm tests performed, about 52% (85/164) showed no significant effects, and 48% (79/164) showed significant effects, excluding the structural gene dosage effects. Of the significant effects, 56% (44/79) were positive effects, and 44% (35/79) were negative effects. Comparisons of dosage effects on orthologous loci (both x-type or both y type HMW glutenins) showed that orthologous HMW glutenin genes are largely influenced by the same regulatory systems. Less correlation was found for comparisons between paralogous genes, although considerable conservation was observed at this level as well. These observations suggest that after polyploidization, many of the duplicated orthologous regulatory loci were inactivated by mutation, thus consolidating control over the HMW glutenin genes. Possible candidates for orthologous regulatory genes were identified in maize and barley. This study represents the first comprehensive search of the wheat genome for regulators of the HMW glutenins. PMID- 12582847 TI - Salt-tolerant mutants in glycophytic salinity response (GSR) genes in Catharanthus roseus. AB - The periwinkle Catharanthus roseus shares glycophytic properties of crop plants. To contribute towards an understanding of the glycophytic response to salinity, large populations of M(2) seeds having an origin in nitroso-methyl urea and ethyl methane sulphonate treatments were screened for germination with 250 mM of NaCl. Out of the nine mutant lines so recovered, which tolerated salt stress due to loss of the normal glycophytic salinity response ( GSR), the characteristics of six gsr mutants are reported here. All six, gsr-1 to gsr-6, differed from the wild-type in both seedling and adult-plant morphological characters beside being salt tolerant. The mutations in them were inherited as monogenic recessive alleles at the corresponding wild-type loci. The trans-complementation tests revealed that the gsr-1 to gsr-6 mutants specified six complementation groups. The mutant seedlings generally accumulated more proline and glycine betaine, constitutively, than the wild-type. The mutant plants transpired lower amounts of water and accumulated higher amounts of proline under drought stress. It was inferred that the products of the six GSR genes defined here are involved in the regulation of salt stress, as well as cell division, developmental and/or morphogenetic pathway(s), in C. roseus. PMID- 12582848 TI - The ploidy level of transgenic plants in Agrobacterium-mediated transformation of tomato cotyledons ( Lycopersicon esculentum Mill.) is genotype and procedure dependent [corrected]. AB - A protocol avoiding the feeder-layer cell system was optimized for Agrobacterium mediated transformation of tomato cotyledonary explants. Over 500 transgenic plants from five tomato cultivars were regenerated in 15 independent experiments. Depending on both genotype and procedure, transformation frequencies ranged from 1.8% to 11.3%. The optimal transformation rate was obtained by inoculating explants with a bacterial suspension in exponential growth ( D(600) = 10(2)-10(3) cells/ml) and transferring cotyledon explants to fresh selective regeneration medium every 3 weeks. The ploidy level of both tomato genotypes used as explant source and primary transformants, was studied by flow cytometry. The inbred lines and cultivars were diploid but a polysomatic pattern in the cotyledon explant was confirmed. The rate of tetraploid transgenic plants ranged from 24.5% to 80% and depended on both the genotype and the transformation procedure. Surprisingly, the percentages of transformed plants with higher ploidy levels were not related to the proportion of 4C and 8C nuclei in the cotyledonary tissue. For some genotypes the optimisation of the transformation rate resulted in an increase of tetraploid transgenic plants. Results obtained in this work indicate the convenience of checking the ploidy level of the primary transformants before performing basic studies or introducing tomato transgenic material in a breeding program. PMID- 12582849 TI - Asymmetry of gene flow and differential geographical structure of molecular diversity in wild and domesticated common bean (Phaseolus vulgaris L.) from Mesoamerica. AB - Using amplified fragment length polymorphisms (AFLPs), we analyzed the genetic structure of wild and domesticated common bean (Phaseolus vulgaris L.) from Mesoamerica at different geographical levels to test the hypothesis of asymmetric gene flow and investigate the origin of weedy populations. We showed both by phenetic and admixture population analyses that gene flow is about three- to four fold higher from domesticated to wild populations than in the reverse direction. This result, combined with other work, points to a displacement of genetic diversity in wild populations due to gene flow from the domesticated populations. The weedy populations appear to be genetically intermediate between domesticated and wild populations, suggesting that they originated by hybridization between wild and domesticated types rather than by escape from cultivation. In addition, the domesticated bean races were genetically similar confirming a single domestication event for the Mesoamerican gene pool. Finally, the genetic diversity of the domesticated bean population showed a lower level of geographic structure in comparison to that of the wild populations. PMID- 12582850 TI - Characterization of expressed NBS-LRR resistance gene candidates from common bean. AB - A complex ancestral resistance (R) gene cluster, localized at the end of linkage group B4, and referred to as the B4 R gene cluster, has been previously genetically characterized. The B4 R gene cluster existed prior to the separation of the two major gene pools of cultivated common bean and contains several resistance specificities effective against the fungus Colletotrichum lindemuthianum. In this paper we report the molecular analysis of four expressed resistance gene candidates (RGCs) that map at the B4 R-cluster and co-localize with R-specificities or R-QTLs effective against C. lindemuthianum. These RGCs have been isolated from two genotypes that are representative of the two major gene pools of common bean: the BA8 and BA11 RGCs originating from the Mesoamerican BAT93 genotype, and the JA71 and JA78 RGCs originating from the Andean JaloEEP558 genotype. These RGCs encode NBS-LRR resistance-like proteins that are closely similar to the tomato I2 R-protein. Based upon sequence comparisons and genetic localization, we established that these four bean RGCs belong to two different subfamilies of R-sequences independently of their gene pool of origin. No feature discriminating the four RGCs according to their gene pool of origin has been observed yet. Comparative sequence analyses of the full length RGCs and their flanking genomic sequences confirmed the ancestral origin of the B4 R-cluster. PMID- 12582851 TI - Molecular tagging of a major QTL for fiber strength in Upland cotton and its marker-assisted selection. AB - Fiber is a basic raw material in the textile industry. The changes in spinning technology have in common the requirement of unique and often greater cotton fiber quality, especially strength, for processing. We used a Gossypium anomalum introgression line, 7235, characterized by good fiber quality properties, to identify molecular markers linked to fiber-strength QTLs. By the use of F(2) and F(3) populations derived from a cross between 7235 and TM-1, a genetic standard of Upland cotton, nine molecular markers, three SSRs and six RAPDs, were identified to be linked to two QTLs for fiber strength. One was a major QTL, QTL(FS1), detected both in Nanjing and Hainan, China, and the Texas College Station, USA. It was found to be associated with eight markers and explained more than 30% of the phenotypic variation. QTL(FS1) was mapped to chromosome 10. The major QTL in 7235 was identified to be transferred from an Acala 3080 cotton. The marker-assisted selection revealed that DNA markers linked to this QTL could be used in increasing the fiber strength of commercial cultivars. PMID- 12582852 TI - A plant-transformation-competent BIBAC library from the Arabidopsis thaliana Landsberg ecotype for functional and comparative genomics. AB - The genome of the model plant Arabidopsis thaliana has been sequenced to near completion. To facilitate experimental determination of the function of every gene in the species, we constructed a large-insert library from the Landsberg ecotype using a plant-transformation-competent binary BAC vector, BIBAC2. The library contains 11,520 clones with an estimated average insert size of 162 kb. Of a sample of 102 clones, 17.6% had no inserts; further, in the library as a whole, 287 clones contained chloroplast DNA, and 25 contained mitochondrial DNA. Thus it is estimated that 9,295 clones originated from the nuclear genome, representing a 11.5 x coverage. The library was further characterized by screening with probes corresponding to 180-bp repeats, 5S rDNA, 18S-25S rDNA and 23 single-copy RFLP markers. The results showed that 92 clones contained 180-bp centromeric repeats, 78 contained 5S rDNA and 95 contained 18S-25S rDNA, approximately 1%, 0.8% and 1%, respectively, of the nuclear clones in the library. Screening the library with the 23 RFLP markers showed that each one hybridized to an average of seven clones. This library is the first large-insert DNA library for the widely studied Landsberg erecta strain. It will greatly facilitate gene identification by complementation screening, and will enhance analysis of the structure, organization and evolution of the A. thaliana genome. PMID- 12582853 TI - Identification of cut rose (Rosa hybrida) and rootstock varieties using robust sequence tagged microsatellite site markers. AB - In this study a DNA fingerprinting protocol was developed for the identification of rose varieties based on the variability of microsatellites. Microsatellites were isolated from Rosa hybrida L. using enriched small insert libraries. In total 24 polymorphic sequenced tagged microsatellite site (STMS) markers with easily scorable allele profiles, from six different linkage groups, were used to characterize 46 Hybrid Tea varieties and 30 rootstock varieties belonging to different species (Rosa canina L., Rosa indica Thory., Rosa chinensis Jacq., Rosa rubiginosa L., and Rosa rubrifolia glauca Pour.). Clones and known flower color mutants were identified as being identical, all other varieties were differentiated by a unique pattern with as few as three STMS markers. The high discriminating power of the loci suggests that a selection of the most-robust STMS markers may be able to differentiate any two varieties within rootstocks or Hybrid Teas except for mutants. The selected STMS markers will be useful as a tool for reference collection management, for assessing essential derivation of varieties and illegal propagation. PMID- 12582854 TI - Tagging and mapping of rice sheath blight resistant gene. AB - Sheath blight (Rhizoctonia solani Kuhn) is one of the severe rice diseases worldwide. In this study, an F(2) population from a cross between "4011" and "Xiangzaoxian19" is used to identify molecular markers linked with the resistant trait. "4011" was a transgenic rice cultivar carrying a resistant gene to sheath blight, while "Xiangzaoxian19" is a highly susceptible one. As a result, five molecular markers, including three RFLP markers converted from RAPD and AFLP markers, and two SSR markers were identified to link with the sheath blight resistant gene. This dominant resistant gene was named as R sb 1 and mapped on rice chromosome 5. The linkage distance between the markers (E-AT:M-CAC(120), E AT:M-CTA(230), OPN-16(2000), RM164(320) and RM39(300)) and R sb 1 was 1.6 cM, 9.9 cM, 1.6 cM, 15.2 cM and 1.6 cM, respectively. PMID- 12582855 TI - Self-incompatibility in passionfruit: evidence of gametophytic-sporophytic control. AB - Self-incompatibility in passionfruit was studied in families originated from crosses among plants that presented differences in reciprocal crosses. The three families, obtained by crossing S(3) plants, exhibited one incompatible group; no reciprocal differences were observed. The phenotype of the families was the same as the parent plants, S(3). These results suggest the presence of a gene ( G), gametophytic in its action, associated to the sporophytic gene S, modifying the incompatibility reaction in passionfruit. The reciprocal difference exhibited in the crosses among the parents could be explained as a matching between plants homozygous for S, but homozygous and heterozygous for G. Actually this would be a partially compatible cross, not detectable when the evaluation is done based on fruit set data. As the family originated from this kind of cross is homozygous for S and heterozygous for G, no reciprocal differences are expected, and the phenotype should be the same as the parental plants, as observed in the present work. PMID- 12582856 TI - Comparative analysis of chloroplast DNA in Pyrus species: physical map and gene localization. AB - A physical map of chloroplast DNA (cpDNA) of pear [Pyrus ussuriensis var. hondoensis (Nakai et Kikuchi) Rehder] was constructed using five restriction enzymes, SalI, XhoI, BamHI, SacI and PstI. This information will make it possible to investigate the phylogenetic relationships between Pyrus species. Pear cpDNA was found to be a circular molecule with a total size of about 156 kb in which two inverted repeats of 24.8 kb divide the molecule into small (17 kb) and large (90 kb) single-copy regions. The endonuclease recognition sites in the physical map were determined by single and double digestion of 13 lambda phage clones which covered the entire sequence of the pear cpDNA. Twenty nine genes were localized on the physical map of the pear cpDNA. The structure of pear cpDNA was almost the same in terms of genome size and gene order as that of tobacco cpDNA. RFLP analysis was carried out on cpDNAs from five Pyrus species (Pyrus pyrifolia, Pyrus ussuriensis, Pyrus calleryana, Pyrus elaeagrifolia and Pyrus communis). Two mutations, a recognition-site mutation and a length mutation (deletion), were found only in the cpDNA of P. pyrifolia cultivars. These mutations were localized on the physical map of pear cpDNA. The number of mutations of cpDNA in Pyrus species are small in comparison with those of other angiosperms, suggesting a high degree of genome conservatism in Pyrus species. PMID- 12582857 TI - Pollen viability restoration in a Coffea canephora P. and C. heterocalyx Stoffelen backcross. QTL identification for marker-assisted selection. AB - Male fertility of interspecific hybrids was analysed in one F1 and two backcrossed progenies originating from a cross between Coffea canephora and Coffea heterocalyx. Male fertility was tested using pollen stainability with acetic carmine. The results showed a marked decline in fertility at the F1 level, and fertility was almost fully restored after two backcrosses. The computed broad sense heritability represented 47% of the variance. Quantitative trait loci (QTLs) locations and effects on pollen viability were estimated using an amplified fragment length polymorphism (AFLP) genetic linkage map constructed in the segregating BC1 population. Three significant QTLs (LOD>3 and p < 0.001 by ANOVA) were detected for pollen viability, two of which were responsible for the bimodal distribution of pollen viability in the segregating population. One QTL was involved in fertility variations among fertile BC1 plants. Fertility inheritance is discussed in relation with previously demonstrated chromosomal sterility in Coffea hybrids and the effect of detected QTLs. The potential use of genetic markers to overcome sterility in interspecific hybrids is also discussed. PMID- 12582858 TI - Can the QTL for late blight resistance on potato chromosome 5 be attributed to foliage maturity type? AB - We investigated the association between late blight resistance and foliage maturity type in potato by means of molecular markers. Two QTLs were detected for foliage resistance against Phytophthora infestans (on chromosomes 3 and 5) and one for foliage maturity type (on chromosome 5). The QTL for resistance to late blight and the QTL for foliage maturity type on chromosome 5 appeared to be mapped on indistinguishable positions. We were interested whether this genetic linkage was due to closely linked but different genes, or due to one (or more) gene(s) with pleiotropic effects. We therefore developed an approach to detect QTLs, in which resistance to late blight was adjusted for foliage maturity type. This analysis revealed the same two QTLs for resistance against P. infestans, but the effect of the locus on chromosome 5 was reduced to only half the original effect. This is a strong indication that the two indistinguishable QTLs for foliage maturity type and for late blight resistance on chromosome 5 may actually be one gene with a pleiotropic effect on both traits. However, there was still a significant effect on resistance against P. infestans on the locus on chromosome 5 after adjusting for foliage maturity type. Therefore we cannot rule out the presence of two closely linked QTLs on chromosome 5: one with a pleiotropic effect on both late blight resistance and foliage maturity type, and another with merely an effect on resistance. In addition, the two QTLs for resistance to late blight showed an important epistatic interaction, suggesting that QTLs for resistance affect each other's expression. PMID- 12582859 TI - Simultaneous improvement for four quality traits of Zhenshan 97, an elite parent of hybrid rice, by molecular marker-assisted selection. AB - "Zhenshan 97" is the female parent of a number of widely used hybrids for rice production in China. However, this line is of poor quality because of a high amylose content (AC), a hard gel consistency (GC) and a low gelatinization temperature (GT), together with a chalky endosperm. It had been determined that the three traits for cooking and eating quality, AC, GC and GT, are controlled by the Waxy locus and/or the tightly linked genomic region. In this study we improved the eating and cooking quality of Zhenshan 97 by introgressing the Waxy gene region from Minghui 63 (wx-MH), a restorer line, that has medium AC, soft GC and high GT. The wx-MH fragment was transferred to Zhenshan 97B by three backcrosses and one selfing, then from Zhenshan 97B to Zhenshan 97A by a cross and a backcross. Molecular marker-assisted selection was applied in the series to select for individuals carrying wx-MH, to identify recombination between the Waxy and flanking markers, and also to recover the genetic background of the recurrent parent. According to the marker genotypes, the improved versions of Zhenshan 97B and Zhenshan 97A, or Zhenshan 97B(wx-MH) and Zhenshan 97A(wx-MH), were the same as the originals except for the Waxy region of less than 6.1 cM in length. The selected lines and their hybrids with Minghui 63, or Shanyou 63(wx-MH), showed a reduced AC and an increased GC and GT, coupled with a reduced grain opacity. Field examinations of agronomic performance revealed that Zhenshan 97B(wx-MH) and Shanyou 63(wx-MH) were essentially the same as the originals except for a significant decrease in grain weight. The simultaneous improvement of AC, GA, GT and opacity, indicated that the Waxy region had major effects on the four quality traits. The improved versions of Zhenshan 97 A and B should be immediately useful in hybrid rice production. PMID- 12582860 TI - Effect of recombination in the parent populations on the means and combining ability variances in hybrid populations of maize ( Zea mays L.). AB - Recombination of selected genotypes plays a key role in plant breeding for generating new base populations. We investigated the influence of recombination in two parent populations on the means and combining ability variances of their hybrid population by (1) quantitative genetic theory and (2) experiments with maize. The two parent populations were founded by four early flint and four early dent inbred lines, respectively. Each population was studied in three generations: Syn-0, the four inbred lines themselves; Syn*-1, the six intrapool single crosses (SC); and Syn*-2, the three intrapool double crosses (DC). Four interpool hybrid populations were created: (1) all 16 SC and (2) all 36 DC were produced from generations Syn-0 and Syn*-1, respectively, (3) 168 biparental progenies (BIP) of type flint x dent (female x male), and (4) 168 BIP of type dent x flint were produced according to NC-design I with randomly sampled plants of generation Syn*-2. The half-sib and full-sib families obtained in this manner were evaluated for grain yield, dry matter concentration and plant height. According to theoretical results, differences in the population means of these hybrid populations indicate the presence of various types of epistasis. Changes in combining ability variances from SC to DC reflect different levels of parental inbreeding (F = 1 vs F = 0), whereas changes from DC to BIP only reflect the effects of recombination and are attributable to covariances between additive and dominance effects caused by linkage disequilibrium in the Syn-0 generations. The experimental results showed a significant decline in yield from DC to BIP due to a loss of gene combinations with favourable epistatic effects. Estimates of sigma(2)(GCA) attributable to flint or dent lines decreased or remained unchanged from SC to DC, but generally increased in the BIP populations. The consequences of these trends for developing improved interpool hybrids are discussed. PMID- 12582862 TI - Genetic variation of oaks ( Quercus spp.) in Switzerland. 3. Lack of impact of postglacial recolonization history on nuclear gene loci. AB - Quercus petraea, Quercus pubescens and Quercus robur are closely related and interfertile white oaks native to Switzerland. The three species are known to share identical cpDNA haplotypes, which are indicative of the postglacial recolonization history of populations. Only two haplotypes are common in Switzerland. We compared variation of cpDNA and of isozymes in 28 oak populations from Switzerland in order to assess the impact of the postglacial population history on current genetic structures of nuclear controlled isozyme gene loci. Species delineation was based on Principal Component Analysis of leaf morphological traits. The species status of populations was reflected at isozyme gene loci, but differentiation between populations with different cpDNA haplotypes and hence different recolonization history was very low at enzyme gene loci for all species. Thus, glacial and postglacial population history was not reflected at nuclear gene loci on the temporal and spatial scale covered by the present study. Extensive gene flow through pollen among populations is likely to have blurred a previously existing genetic differentiation at biparentally inherited gene loci that possibly evolved in the different glacial refugia of the above mentioned cpDNA haplotypes. PMID- 12582861 TI - Chromosomal location of a Triticum dicoccoides-derived powdery mildew resistance gene in common wheat by using microsatellite markers. AB - The powdery mildew resistance has been transferred from an Israeli wild emmer (Triticum dicoccoides) accession "G-305-M" into common wheat by crossing and backcrossing (G-305-M/781//Jing 411*3). Genetic analysis showed that the resistance was controlled by a single dominant gene at the seedling stage. Among the 102 pairs of SSR primers tested, four polymorphic microsatellite markers (Xpsp3029, Xpsp3071, Xpsp3152 and Xgwm570) from the long arm of chromosome 6A were mapped in a BC(2)F(3) population segregating for powdery mildew resistance and consisting of 167 plants. The genetic distances between the resistance gene and these four markers were: 0.6 cM to Xpsp3029, 3.1 cM to Xpsp3071, 11.2 cM to Xpsp3152 and 20.4 cM to Xgwm570, respectively. The order of these microsatellite loci agreed well with the established microsatellite map of chromosome arm 6AL. We concluded that the resistance gene was located on the long arm of chromosome 6AL. Based on the origin and chromosomal location of the gene, it is suggested that the resistance gene derived from "G-305-M" is a novel powdery mildew resistance gene and is temporarily designated MlG. PMID- 12582863 TI - Localisation of genes for resistance against Blumeria graminis f.sp. hordei and Puccinia graminis in a cross between a barley cultivar and a wild barley (Hordeum vulgare ssp. spontaneum) line. AB - The aims of this investigation have been to map new (quantitative) resistance genes against powdery mildew, caused by Blumeria graminis f.sp. hordei L., and leaf rust, caused by Puccinia hordei L., in a cross between the barley ( Hordeum vulgare ssp. vulgare) cultivar "Vada" and the wild barley ( Hordeum vulgare ssp. spontaneum) line "1B-87" originating from Israel. The population consisted of 121 recombinant inbred lines. Resistance against leaf rust and powdery mildew was tested on detached leaves. The leaf rust isolate "I-80" and the powdery mildew isolate "Va-4", respectively, were used for the infection in this experiment. Moreover, powdery mildew disease severity was observed in the field at two different epidemic stages. In addition to other DNA markers, the map included 13 RGA (resistance gene analog) loci. The structure of the data demanded a non parametric QTL-analysis. For each of the four observations, two QTLs with very high significance were localised. QTLs for resistance against powdery mildew were detected on chromosome 1H, 2H, 3H, 4H and 7H. QTLs for resistance against leaf rust were localised on 2H and 6H. Only one QTL was common for two of the powdery mildew related traits. Three of the seven QTLs were localised at the positions of the RGA-loci. Three of the five powdery mildew related QTLs are sharing their chromosomal position with known qualitative resistance genes. All detected QTLs behaved additively. Possible sources of the distorted segregation observed, the differences between the results for the different powdery mildew related traits and the relation between qualitative and quantitative resistance are discussed. PMID- 12582864 TI - Development and characterization of simple sequence repeat (SSR) markers and their use in determining relationships among Lycopersicon esculentum cultivars. AB - The simple sequence repeat (SSR) or microsatellite marker is currently the preferred molecular marker due to its highly desirable properties. The aim of this study was to develop and characterize more SSR markers because the number of SSR markers currently available in tomato is very limited. Five hundred DNA sequences of tomato were searched for SSRs and analyzed for the design of PCR primers. Of the 158 pairs of SSR primers screened against a set of 19 diverse tomato cultivars, 129 pairs produced the expected DNA fragments in their PCR products, and 65 of them were polymorphic with the polymorphism information content (PIC) ranging from 0.09 to 0.67. Among the polymorphic loci, 2-6 SSR alleles were detected for each locus with an average of 2.7 alleles per locus; 49.2% of these loci had two alleles and 33.8% had three alleles. The vast majority (93.8%) of the microsatellite loci contained di- or tri-nucleotide repeats and only 6.2% had tetra- and penta-nucleotide repeats. It was also found that TA/AT was the most frequent type of repeat, and the polymorphism information content (PIC) was positively correlated with the number of repeats. The set of 19 tomato cultivars were clustered based on the banding patterns generated by the 65 polymorphic SSR loci. Since the markers developed in this study are primarily from expressed sequences, they can be used not only for molecular mapping, cultivar identification and marker-assisted selection, but for identifying gene trait relations in tomato. PMID- 12582865 TI - Pyramiding transgenes for multiple resistance in rice against bacterial blight, yellow stem borer and sheath blight. AB - Here we describe the development of transgene-pyramided stable elite rice lines resistant to disease and insect pests by conventional crossing of two transgenic parental lines transformed independently with different genes. The Xa21 gene (resistance to bacterial blight), the Bt fusion gene (for insect resistance) and the chitinase gene (for tolerance of sheath blight) were combined in a single rice line by reciprocal crossing of two transgenic homozygous IR72 lines. F4 plant lines carrying all the genes of interest stably were identified using molecular methods. The identified lines, when exposed to infection caused by Xanthomonas oryzae pv oryzae, showed resistance to bacterial blight. Neonate larval mortality rates of yellow stem borer ( Scirpophaga incertulas) in an insect bioassay of the same identified lines were 100%. The identified line pyramided with different genes to protect against yield loss showed high tolerance of sheath blight disease caused by Rhizoctonia solani. PMID- 12582866 TI - Genetic linkage maps constructed by using an interspecific cross between Japanese and European pears. AB - Genetic linkage maps of the European pear ( Pyrus communis L.) cultivar 'Bartlett' and the Japanese pear ( Pyrus pyrifolia Nakai) cultivar 'Housui' were constructed based on AFLPs, SSRs from pear, apple and Prunus, isozymes and phenotypic traits by using their F(1) progenies. The map of the female parent Bartlett consisted of 226 loci including 175 AFLPs, 49 SSRs, one isozyme and one S locus on 18 linkage groups over a total length of 949 cM, while that for 'Housui' contained 154 loci including 106 AFLPs, 42 SSRs, two phenotypic traits and the other four markers on 17 linkage groups encompassing a genetic distance of 926 cM. These maps were partially aligned using 20 codominant markers which showed segregating alleles in both parents. Compared with the reports of apple genetic maps, these pear maps were not saturated but were near saturation. Distorted segregation was observed in two and one regions of the genome of Bartlett and Housui, respectively. The position of 14 SSRs originating from apple could be successfully determined in pear maps, which enabled us to compare the two maps. Some SSRs developed from Prunus (peach, cherry) were also mapped. The relationships between pear and the other species belonging to the Rosaceae were discussed based on the position of SSRs. PMID- 12582867 TI - Mapping QTLs for grain hardness and puroindoline content in wheat ( Triticum aestivum L.). AB - Genes for puroindoline-a (Pin-a), puroindoline-b (Pin-b) and grain-softness proteins (GSP) have been shown to be linked to the dominant Ha locus responsible for the soft texture of the grain. Though linkage has been demonstrated of the puroindoline genes to the Ha locus, there is no clear evidence that puroindoline content is the product of the gene Ha. A segregating population of 115 recombinant inbred lines (RILs) originating from a cross between the hexaploid Synthetic wheat ( Triticum durum x Aegilops tauschii, W 7984) and the cultivar 'Opata' (M 85) was studied in two different experimental years to detect Quantitative Trait Loci (QTLs) for three traits: grain hardness (Hard), puroindoline-a (Pin-a) and puroindoline-b (Pin-b) contents. The detection of QTLs was performed using marker linear regression. Negative correlation coefficients ( 0.86 and -0.80) were identified between grain hardness and puroindoline content (a and b, respectively) on data obtained in 1996. Results obtained in 1999 confirmed the negative correlation between Hard and Pin-a (-0.73); however a positive correlation coefficient was found with Pin-b content (0.41). Total phenotypic variation explained by each QTL was calculated (R2). For each of the Hard, Pin-a and Pin-b traits one major QTL was detected on the short arm of chromosome 5D, located close to the mta9 allele (puroindoline-a). For the first year (1996) the QTL in this region explained around 63% of the phenotypic variability in grain hardness, 77% in Pin-a and 45% in Pin-b contents. These values were confirmed in trials carried out in 1999 with a R2 value of 0.71, 0.72 and 0.25 for Hard, Pin-a and Pin-b, respectively. In 1996 and 1999 a second major QTL was detected for grain hardness on the long arm of the same chromosome. Present results indicate that it cannot be definitely concluded that puroindoline content represents a linear explanation for variations in grain hardness. PMID- 12582868 TI - Quantitative trait loci for partial resistance to Aphanomyces root rot in pea. AB - Aphanomyces root rot, caused by Aphanomyces euteiches Drechs, is the most important disease of pea ( Pisum sativum L.) worldwide. No efficient chemicals are available to control the pathogen. To facilitate breeding for Aphanomyces root rot resistance and to better understand the inheritance of partial resistance, our goal was to identify QTLs associated with field partial resistance. A population of 127 RILs from the cross Puget (susceptible) x 90-2079 (partially resistant) was used. The lines were assessed for resistance to A. euteiches under field conditions at two locations in the United States (Pullman, Wash. and LeSueur, Minn.) in 1996 and 1998 for three criteria based on symptom intensity and disease effects on the whole plant. The RILs were genotyped using automated AFLPs, RAPDs, SSRs, ISSRs, STSs, isozymes and morphological markers. The resulting genetic map consisted of 324 linked markers distributed over 13 linkage groups covering 1,094 cM (Kosambi). Twenty seven markers were anchored to other published pea genetic maps. A total of seven genomic regions were associated with Aphanomyces root rot resistance. The first one, located on LG IVb and named Aph1, was considered as "major" since it was highly consistent over the years, locations and resistance criteria studied, and it explained up to 47% of the variation in the 1998 Minnesota trial. Two other year-specific QTLs, namely Aph2 and Aph3, were revealed from different scoring criteria on LG V and Ia, respectively. Aph2 and Aph3 mapped near the r (wrinkled/round seeds) and af (normal/afila leaves) genes, and accounted for up to 32% and 11% of the variation, respectively. Four other "minor" QTLs, identified on LG Ib, VII and B, were specific to one environment and one resistance criterion. The resistance alleles of Aph3 and the two "minor" QTLs on LG Ib were derived from the susceptible parent. Flanking markers for the major Aphanomyces resistance QTL, Aph1, have been identified for use in marker-assisted selection to improve breeding efficiency. PMID- 12582869 TI - Trinucleotide microsatellites in Norway spruce ( Picea abies): their features and the development of molecular markers. AB - Trinucleotide microsatellites have proven to be the markers of choice in human genetic analysis because they are easier to genotype than dinucleotides. Their development can be more time-consuming due to their lower abundance in the genome. We isolated trinucleotide microsatellites in Norway spruce ( Picea abies K.) using an enrichment procedure for the genomic-library construction. Here we report on the characterisation of 85 ATC microsatellite-containing clones, from which 39 markers were developed. Many of the clones showed the occurrence of tandem repeats of higher order than the trinucleotide ones, often resembling minisatellite repeats. The sequencing of a sample of the alleles at one of the loci revealed size homoplasy due to base substitutions within the microsatellite region. The presence of ATC motifs within repetitive sequence families was observed. We found a significant relationship between the level of polymorphism and the length of the microsatellite. The levels of variability for ATC trinucleotide markers were lower than those for dinucleotides, both when tested on all loci in a set of six individuals and on a subset of loci in four natural populations. This difference is most likely attributable to lower mutation rates for trinucleotide than for dinucleotide loci. The availability of markers with different mutation rates allows one to select the proper marker set to investigate population processes on different time scales. PMID- 12582870 TI - Transgenics of an elite indica rice variety Pusa Basmati 1 harbouring the codA gene are highly tolerant to salt stress. AB - Transgenic lines of indica rice were generated by Agrobacterium-mediated transformation with the choline oxidase ( codA) gene from Arthrobacter globiformis. Choline oxidase catalyses conversion of choline to glycine betaine. Glycine betaine is known to provide tolerance against a variety of stresses. Molecular analyses of seven independent transgenic lines as performed by Southern, Northern and Western hybridization revealed integration and expression of the transgene as well as inheritance in the progeny plants. A good correlation was observed between levels of mRNA and protein accumulation, and a significant amount of choline oxidase product, i.e. glycine betaine, accumulated in R0 as well as R1 plants. Mendelian as well as non-Mendelian segregation patterns were obtained in the progeny plants. Challenge studies performed with R1 plants by exposure to salt stress (0.15 M NaCl) for 1 week, followed by a recovery period, revealed that in some cases more than 50% of the transgenic plants could survive salt stress and set seed whereas wild-type plants failed to recover. PMID- 12582871 TI - Genetic analysis and QTL mapping of early root bulking in an F1 population of non inbred parents in cassava ( Manihot esculenta Crantz). AB - The genetic basis of early bulking in cassava was studied in a replicated, multi locational trial using 144 F1 progeny derived from an intra-specific cross between two non-inbred parents. A second, sequential harvest experiment examined the relative importance of eight yield-related traits on early bulking and their QTLs during the crop growth cycle. Our objectives were to identify traits, and genes controlling them, strongly associated with early yield as a first step to marker-assisted improvement of the trait. Multiple linear regression analysis and stepwise regression of early yield on eight yield-related traits revealed harvest index, dry foliage weight and root diameter as the most important factors associated with early yield. A total of 18 QTLs controlling early yield were identified in the first and second experiments and 27 QTLs, 2 for dry foliage weight, 8 for harvest index and 17 for root diameter, in the second experiment. The individual effects of alleles at these QTLs identified ranged from 7% to 33% of the phenotypic variance explained. Seven of 18 QTLs found for early yield (39%) coincided with QTLs associated with one or more traits with significant influence on early yield. The results show that sink and source capacities are very important in determining early yield. The identification of a number of QTLs with positive effect for increased early yield provides an opportunity for marker assisted selection and improvement of early bulking potential in cassava. PMID- 12582872 TI - Construction and analysis of a microsatellite-based database of European wheat varieties. AB - A database of 502 recent European wheat varieties, mainly of winter type, was constructed using 19 wheat microsatellites and one secalin-specific marker. All datapoints were generated in at least two laboratories using different techniques for fragment analysis. An overall level of >99.5% accuracy was achieved. The 199 alleles detected allowed discrimination between all of the varieties except duplicates, and varieties derived from identical parents. Approximately 25% of the varieties showed some heterogeneities, with the highest level of heterogeneity in south-eastern European material. The highest genetic diversity and the highest number of rare alleles were found in varieties from southern Europe. The relative allele frequencies varied for most microsatellites in different geographical regions. PMID- 12582873 TI - Engineering deoxynivalenol metabolism in wheat through the expression of a fungal trichothecene acetyltransferase gene. AB - Fusarium head blight occurs in cereals throughout the world and is especially important in humid growing regions. Fusarium head blight (FHB) has re-emerged as a major disease of wheat and barley in the U.S. and Canada since 1993. The primary causal agents of FHB, Fusarium graminearum and Fusarium culmorum, can produce deoxynivalenol (DON), a trichothecene mycotoxin that enhances disease severity and poses a health hazard to humans and monogastric animals. To reduce the effects of DON on wheat, we have introduced FsTRI101, a Fusarium sporotrichioides gene formerly known as TriR, into the regenerable cultivar Bobwhite. TRI101 encodes an enzyme that transfers an acetyl moiety to the C3 hydroxyl group of trichothecenes. Four different transgenic plants carrying the FsTRI101 gene were identified. Although expression levels varied among the four lines, all of them accumulated FsTRI101 transcripts in endosperm and glume. TRI101-encoded acetyltransferase activity was detected in endosperm extracts of a single plant that accumulated FsTRI101 mRNA. Greenhouse resistance tests indicated that the accumulation of FsTRI101-encoded acetyltransferase in this plant confers partial protection against the spread of F. graminearum in inoculated wheat heads (spikes). PMID- 12582874 TI - Microsatellite analysis of wheat chromosome 2D allows the reconstruction of chromosomal inheritance in pedigrees of breeding programmes. AB - The dwarfing gene Rht8 and the photoperiodic insensitivity gene Ppd-D1 are linked on the short arm of chromosome 2D of bread wheat and play an important role in determining the geographic adaptation of modern wheat varieties. The genes are believed to originate from the old Japanese variety 'Akakomugi' and have been distributed throughout the world by diverse breeding programmes. Twelve microsatellite loci previously mapped on wheat chromosome 2D were used for a retrospective analysis of 59 wheat varieties with known pedigree, to trace the transmittance of the chromosomal region around these genes during extended breeding programmes. Within the range of the screened varieties 100 alleles were detected at the 12 microsatellite loci. For each microsatellite locus, a screen over varieties was performed to find the alleles corresponding to the parental variety 'Akakomugi'. A comparison of wheat varieties carrying the 192-bp allele, at locus Xgwm261-2D which is diagnostic for the presence of the gene Rht8, with the varieties without Rht8, showed linkage disequilibrium of 'Akakomugi' alleles for a segment of chromosome 2D which comprised at least 28 cM. Selection was accompanied with a linkage drag of 'Akakomugi' alleles in the neighbouring loci to Rht8. A diminution of the segment of chromosome 2D originating from 'Akakomugi' during several pedigree generations was observed. Varieties of the early generations were found to carry the whole short arm of chromosome 2D of 'Akakomugi', while the varieties of further selections possessed smaller segments including the diagnostic allele at locus Xgwm261-2D. Our results demonstrate that microsatellites can be successfully used for studying the inheritance of chromosomes within pedigrees of breeding programmes. PMID- 12582875 TI - Use of fluorescence in situ hybridization for gross mapping of transgenes and screening for homozygous plants in transgenic barley ( Hordeum vulgare L.). AB - Introduced transgenes, uidA, sgfp (S65T) and/or bar, were localized using fluorescence in situ hybridization (FISH) on metaphase chromosomes of transgenic barley produced by microparticle bombardment of immature embryos. Of the 19 independent transgenic lines (eight diploid and 11 tetraploid), nine had uidA and ten had s gfp (S65T). All lines tested had three or more copies of the transgenes and 18 out of 19 lines had visibly different integration sites. At a gross level, it appeared that no preferential integration sites of foreign DNA among chromosomes were present in the lines tested; however, a distal preference for transgene integration was observed within the chromosome. In diploid T0 plants that gave a 3:1 segregation ratio of transgene expression in the T1, only single integration sites were detected on one of the homologous chromosomes. Homozygous diploid plants had doublet signals on a pair of homologous chromosomes. All tetraploid T0 plants that gave a 3:1 segregation ratio in the T1 generation had only a single integration site on one of the homologous chromosomes. In contrast, the single tetraploid T0 plant with a 35:1 segregation ratio in the T1 generation had doublet signals on a pair of homologous chromosomes. In the one tetraploid T0 line, which had a homozygote-like segregation ratio (45:0), there were doublet signals at two loci on separate chromosomes. We conclude that the application of FISH for analysis of transgenic plants is useful for the gross localization of transgene(s) and for early screening of homozygous plants. PMID- 12582877 TI - Molecular markers for sex determination in papaya ( Carica papaya L.). AB - We have developed molecular markers tightly linked to Sex1, the gene that determines plant sex in papaya ( Carica papaya L.). Three RAPD products have been cloned and a portion of their DNA sequenced. Based on these sequences SCAR primers were synthesized. SCAR T12 and SCAR W11 produce products in hermaphrodite and male plants and only rarely in females. SCAR T1 produces a product in all papayas regardless of plant sex. SCAR T12 and SCAR W11 showed no recombination in a population of 182 F2 plants from a 'SunUp' by 'Kapoho' cross. Based on these results a PCR-based technique for rapidly and accurately determining the sex of papaya plants was developed using either W11 or T12 to detect the hermaphrodite or male allele and T1, which amplifies a product regardless of sex type, as a positive control. The sexing technique, using SCAR T12 and SCAR T1 as a positive control, was used to correctly predict hermaphrodite papaya plants in a population of seedlings with an overall accuracy of 99.2%. PMID- 12582876 TI - Genetic differentiation of wild relatives of rice as assessed by RFLP analysis. AB - To study genetic diversity and relationships of wild relatives of rice, 58 accessions of Oryza rufipogon, Oryza nivara, Oryza sativa f. spontanea and the cultivated Oryza sativa, representing a wide range of their distribution, were analyzed using the restriction fragment length polymorphism (RFLP) technique. All 30-used RFLP probes detected polymorphisms among the Oryza accessions, with an average of 3.8 polymorphic fragments per probe. Considerable genetic diversity was scored among the Oryza accessions with a similarity coefficient ranging from 0.28 to 0.93; but the cluster analysis of the accessions did not show an apparent grouping based on the species classification, instead they were scattered randomly in different groups. Noticeably, the Oryza accessions from the same geographic region, or near-by geographic regions, tended to be clustered in the same groups. The indica rice varieties showed relatively high genetic diversity and were scattered in different groups of their wild relatives, but the japonica varieties showed a relatively low variation and formed an independent group. It is concluded from the molecular analytical result that: (1) the four Oryza taxa have a remarkably close relationship and their independent species status need to be carefully reviewed; (2) geographic isolation has played a significant role in the differentiation of the Oryza accessions; therefore, a wide geographic range needs to be covered in collecting wild rice germplasm for ex situ conservation; and (3) the conventional conclusion of indica rice being directly domesticated from its ancestral wild species, and japonica rice being derived from indica, gains support from our data. PMID- 12582878 TI - An estimation of the minimum number of SSR alleles needed to reveal genetic relationships in wheat varieties. I. Information from large-scale planted varieties and cornerstone breeding parents in Chinese wheat improvement and production. AB - The genetic relationships of 43 wheat varieties were analyzed with SSR markers. The materials employed included 14 cornerstone breeding parents used before 1980 and another 29 other large-scale planted varieties currently in use in China. A total of 501 different alleles were amplified, including 166 alleles of the A genome, 174 of the B genome and 161 of the D genome. Data obtained were used to estimate genetic similarity using the DICE coefficient, and dendrograms were constructed using the UPGMA method. The dendrogram with 501 alleles successfully differentiated all the cornerstone breeding parents and the large-scale planted varieties, and the dendogram tree was basically consistent with the pedigrees of these varieties. The correlation coefficient between the genetic distance matrix of 501 alleles and that of 450 was 0.99. Correlation coefficients among random samples of alleles suggested that 350 to 400 alleles were needed to detect genetic relationships among common wheat varieties. Correlation coefficients of a genetic similarity matrix based on 580, and those of 501 and 400, random alleles were 0.96 and 0.94, respectively. However, there were marked differences between the matrix based on the 501 alleles and those based on markers located on the A-, B- or D-genome independently. The correlation coefficients between the genetic distance matrix of 501 alleles and alleles within A, B or D genomes were 0.77, 0.76 and 0.67. The estimation of genetic similarity should be based on data from all genomes rather than any one genome. PMID- 12582879 TI - QTLs for grain carbon isotope discrimination in field-grown barley. AB - In several crops including cereals, carbon isotope discrimination (Delta) has been associated with drought tolerance in terms of water-use efficiency and yield stability in drought-prone environments. By using a complete genetic map generated from 167 recombinant inbred lines from a cross between Tadmor and Er/Apm, QTLs associated with grain Delta have been detected in barley grown in three Mediterranean field environments, two differing only in water availability. Ten QTLs were identified: one was specific to one environment, two presented interaction with the environment, six presented main effects across three or two environments and one presented both effects. Heading date did not contribute to the environment (E) and G x E effects acting on Delta. Seasonal rainfall and the ratio of rainfall to evapo-transpiration made large contributions to the environmental effect, but their influence on G x E was weaker. Eight QTLs for Delta co-located with QTLs for physiological traits related to plant water status and/or osmotic adjustment, and/or for agronomic traits previously measured on the same population. Some perspectives in terms of characterising drought tolerance are evoked. PMID- 12582880 TI - The origin of Russian cultivars of red clover ( Trifolium pratense L.) and their genetic relationships to wild populations in the Urals. AB - Propagation and breeding of red clover ( Trifolium pratense L.) in Russia was initiated about 200 years ago but the origin of present-day cultivars is disputed. Some authors argue that most Russian cultivars were derived from western European ones, whereas others support a Russian origin of the cultivars from local wild populations. In the present study we assessed the genetic variation at 17 allozyme loci in seven Russian cultivars, bearing the names of localities of the Urals, two American ones that have been used in Russia for scientific experiments and seven wild populations of the Urals and Western Siberia. Variation at the 17 protein loci supports the western European origin of the cultivars and also indicates that gene flow between cultivars and wild populations was limited or has not acted sufficiently long to affect the genetic composition of the red clover wild populations of the Urals. PMID- 12582881 TI - QTL mapping of stay-green in two sorghum recombinant inbred populations. AB - The stay-green trait is a reported component of tolerance to terminal drought stress in sorghum. To map quantitative trait loci (QTLs) for stay-green, two sorghum recombinant inbred populations (RIPs) of 226 F(3:5) lines each were developed from crosses (1) IS9830 x E36-1 and (2) N13 x E36-1. The common parental line, E36-1 of Ethiopian origin, was the stay-green trait source. The genetic map of RIP 1 had a total length of 1,291 cM, with 128 markers (AFLPs, RFLPs, SSRs and RAPDs) distributed over ten linkage groups. The map of RIP 2 spanned 1,438 cM and contained 146 markers in 12 linkage groups. The two RIPs were evaluated during post-rainy seasons at Patancheru, India, in 1999/2000 (RIP 2) and 2000/2001 (RIP 1). The measures of stay-green mapped were the green leaf area percentages at 15, 30 and 45 days after flowering (% GL15, % GL30 and % GL45, respectively). Estimated repeatabilities for % GL15, % GL30 and % GL45 amounted to 0.89, 0.81 and 0.78 in RIP 1, and 0.91, 0.88 and 0.85 in RIP 2, respectively. The number of QTLs for the three traits detected by composite interval mapping ranged from 5 to 8, explaining 31% to 42% of the genetic variance. In both RIPs, both parent lines contributed stay-green alleles. Across the three measures of the stay-green trait, three QTLs on linkage groups A, E and G were common to both RIPs, with the stay-green alleles originating from E36-1. These QTLs were therefore consistent across the tested genetic backgrounds and years. After QTL validation across sites and verification of the general benefit of the stay-green trait for grain yield performance and stability in the target areas, the corresponding chromosomal regions could be candidates for marker assisted transfer of stay-green into elite materials. PMID- 12582882 TI - Ancestors of white clover ( Trifolium repens L.), as revealed by isozyme polymorphisms. AB - Isozymes were used to study the putative ancestors of white clover ( Trifolium repens L.). Ten enzymes were examined, and 18 loci were resolved via starch-gel electrophoresis for accessions representing T. repens, Trifolium isthmocarpum, Trifolium nigrescens, Trifolium occidentale and Trifolium uniflorum, in addition to two more distantly related species, Trifolium alpinum and Trifolium purseglovei. Nei's genetic identities indicate that T. uniflorum and T. nigrescens are the closest relatives of T. repens. The isozyme data thus support a hypothesis that the two genomes of the tetraploid T. repens could have been derived from hybridization between T. nigrescens and T. uniflorum. This conclusion is further supported by shared alleles between T. repens, T. nigrescens and T. uniflorum. However, the origin of T. repens is somewhat obscured by the presence of shared alleles between T. repens and both T. occidentale and T. isthmocarpum, suggesting that introgression of genes from the latter two species into T. repens may also have taken place. High values of genetic identity are shared between T. occidentale with T. nigrescens and T. uniflorum, also indicative of introgression. Alternatively the presence of shared alleles among the five species may reflect their recent common ancestry. PMID- 12582883 TI - Mapping and analysis of quantitative trait loci for grain oil content and agronomic traits using AFLP and SSR in sunflower ( Helianthus annuus L.). AB - Crosses were made between two inbred lines of sunflower. Parents and 118 F(3) families were planted in the field in a randomized complete block design in two replications. Genetic control for some agronomical traits: grain weight by plant (GWP), 1,000-grain weight (TGW), percentage of oil in grain (POG) and sowing to flowering date (STF) was investigated in F(3) families and their parents. Genetic variability was observed among the 118 F(3) families for all the traits studied. Genetic gain was obtained when the best F(3) family, or the mean of 10% of the selected families was compared with the best parent for GWP, TWG and POG. Heritability was 0.23 for GWP, 0.55 for TGW, 0.57 for POG and 0.32 for STF. A set of 244 F(3) families from the same cross, including the above 118 mentioned families and their two parents, were screened with 276 AFLP and microsatellite markers and a linkage map was constructed based on 170 markers. Two putative QTLs for the GWP trait ( gmp), one QTL for TGW ( tgw), six QTLs for POG ( pog) and two for STF ( stf) were detected. The percentage of phenotypic variance explained by each QTL ranged from 2.6% to 70.9%. The percentage of total phenotypic variance explained was 50.7% for GWP, 5.4% for TGW, 90.4% for POG and 89.3% for STF. Although these regions need to be more-precisely mapped, the information obtained should help in marker-assisted selection. PMID- 12582884 TI - Influence of number and map distribution of AFLP markers on similarity estimates in carrot. AB - When genetic diversity among organisms was measured with molecular markers, the question of genome coverage was currently stressed out. In order to check if well distributed, mapped AFLP markers were more efficient in assessing varietal identification of carrot accessions than randomly chosen markers, nine closely related genotypes were analysed. A software was developed to realise 1,000 random choices of 20 to 70 mapped or unmapped markers, offering numerous genome coverages. We statistically showed that taking into account marker position does not provide a better estimation of genetic distances. Moreover, in the case of carrot, we concluded that 60 AFLP markers offer the best compromise between the level of precision and minimal expense. PMID- 12582885 TI - Resistance gene analogs are candidate markers for disease-resistance genes in grape ( Vitis spp.). AB - A set of NBS-containing sequences was isolated from genomic DNA of two grape species ( Vitis amurensis and Vitis riparia) and characterised in a panel of Vitis genotypes carrying different levels of resistance against downy mildew and other diseases. A PCR-mediated approach made use of degenerate primers designed on conserved regions encoding known R-genes, and provided the source for cloning grape analogous sequences. Cloned sequences were digested with ten endonucleases and 29 out of 71 putative recombinant clones, which showed unique restriction patterns, were sequenced. Using a threshold value of 40% identity, at least 12 grape NBS-sequences had a high overall similarity with known R-genes, such as the Arabidopsis gene RPS5 and the tobacco gene N. The presence of internal conserved motifs provided evidence that sequences isolated from grape may belong to the NBS LRR gene family. A cluster analysis based on the deduced amino acid sequence and carried out on grape NBS-sequences, together with several analogous domains of known R-genes, classified grape sequences into three major groups. A grape sequence of each group was used as a probe on Southern blots with digested genomic DNA from resistant and susceptible grapes. One of the NBS-containing probes showed a clear-cut separation between resistant species and susceptible varieties. This evidence makes the probe a candidate marker for disease resistance genes in Vitis germplasm. PMID- 12582886 TI - Assessment of population genetic structure in common wild rice Oryza rufipogon Griff. using microsatellite and allozyme markers. AB - The genetic structure of five natural populations of common wild rice Oryza rufipogon Griff. from China, was investigated with 21 microsatellite loci and compared to estimates of genetic diversity and genetic differentiation detected by 22 allozyme loci. Microsatellite loci, as expected, have much higher levels of genetic diversity (mean values of A = 3.1, P = 73.3%, Ho = 0.358 and He = 0.345) than allozyme loci (mean values of A = 1.2, P = 12.7%, Ho = 0.020 and He = 0.030). Genetic differentiation detected by microsatellite loci ( FST = 0.468, mean I = 0.472) was higher than that for allozyme loci ( FST =0.388, mean I = 0.976). However, microsatellite markers showed less deviation from Hardy-Weinberg expectation (Wright's inbreeding coefficient FIS = -0.069) than do allozymes ( FIS = 0.337). These results suggest that microsatellite markers are powerful high resolution tools for the accurate assessment of important parameters in population biology and conservation genetics of O. rufipogon, and offer advantages over allozyme markers. PMID- 12582887 TI - Inheritance of alleles for glutelin alpha-2 subunit genes in rice and identification of their corresponding cDNA clone. AB - Rice glutelins consist of acidic (alpha) and basic (beta) subunits which are further separated into three polypeptide components assigned as alpha-1, alpha-2, and alpha-3 subunit components and beta-1, beta-2 and beta-3 subunit components. Nine rice mutant lines with a decreased amount of the glutelin alpha-2 subunit component (alpha-2L) were obtained by screening about 6,800 potential mutant lines derived from the fertilized egg treatment with N-methyl-N-nitrosourea (MNU) using the SDS-PAGE method. The mutants were classified into three types of the increased alpha-1 subunit (alpha-1H/alpha-2L), the decreased beta-2 subunit (beta 2L/alpha-2L) and the increased alpha-3 subunit (alpha-3H/alpha-2L) represented by EM278, CM1707 and EM659, respectively. Iso-electric focus (IEF) analysis revealed that all of the mutants had an extremely low amount of a polypeptide with a 6.71 pI value, whereas a polypeptide with either a 6.50 pI value or with a 6.90 pI value increased significantly in alpha-1H/alpha-2L mutants or in alpha-3H/alpha 2L mutants, respectively. The beta-2L/alpha-2L mutants had a decreased amount of a basic polypeptide with a 8.74 pI value. Genetic analysis revealed that the three types of mutants were controlled by a single incomplete dominant gene respectively, and the three are alleles. The gene was temporarily named glu4, which was found to be located on chromosome 1 linked with the eg and spl6 genes. Two-dimensional electrophoresis analysis revealed that the glu4 encoded polypeptides of pI 6.71/alpha-2 and pI 8.74/beta-2. Amino acid sequence analysis suggested that the mutated acidic polypeptide was the product of a GluA subfamily gene. Northern and RT-PCR analyses revealed that glu4 corresponded to the GluA-1 gene. PMID- 12582888 TI - Reduction of species in the wild potato Solanum section Petota series Longipedicellata: AFLP, RAPD and chloroplast SSR data. AB - Species boundaries were assessed with three molecular markers [AFLPs, RAPDs and chloroplast simple sequence repeats (cpSSRs)] for all six species of wild potatoes ( Solanum section Petota) assigned to ser. Longipedicellata: Solanum fendleri, S. hjertingii, S. matehualae, S. papita, S. polytrichon and S. stoloniferum. These tetraploid (2n = 4 x = 48) species grow in the southeastern United States ( S. fendleri) and Mexico (all six species), and a recent morphological analysis supported only three species: (1) S. polytrichon, (2) S. hjertingii (including S. matehualae) and (3) S. stoloniferum (including S. fendleri and S. papita). We analyzed all six species of ser. Longipedicellata (tetraploid) and also analyzed diploids in ser. Bulbocastana, ser. Pinnatisecta, ser. Polyadenia and ser. Tuberosa; tetraploids in ser. Acaulia and hexaploids in ser. Demissa. Concordant with morphological data, AFLP and RAPD results support the synonymy of S. hjertingii and S. matehualae, and completely intermix S. papita and S. fendleri. However, accessions of S. stoloniferum have a tendency to cluster but with exceptions, and S. polytrichon is completely intermixed with S. fendleri and S. papita. The cpSSRs fail to distinguish any of the species in ser. Longipedicellata. Combined morphological and molecular data support only two species in ser. Longipedicellata: S. hjertingii and S. stoloniferum. PMID- 12582889 TI - A pyramid of loci for partial resistance to Fusarium solani f. sp. glycines maintains Myo-inositol-1-phosphate synthase expression in soybean roots. AB - Myo-inositol 1-phosphate synthase (MIPS; EC 5.5.1.4) converts glucose 6-phosphate to myo-inositol 1-phosphate in the presence of NAD(+). It catalyzes the first step in the synthesis of myo-inositol and pinitol, and is a rate limiting step in the de novo biosynthesis of inositol in eukaryotes. Therefore, MIPS is involved in biotic and abiotic stress via Ca(2+) signalling. Seedlings of four soybean genotypes were inoculated with Fusarium solani f. sp. glycines, the causative agent of sudden death syndrome (SDS), and differentially abundant mRNAs were identified by differential display. The genotypes carried either zero, two, four or six alleles of the quantitative trait loci (QTLs) that control resistance to SDS in an additive manner. The mRNA abundance of MIPS did not decrease following inoculation in a recombinant inbred line (RIL 23) containing all six resistance alleles of the QTLs conferring resistance to SDS of soybean. However, the abundance of MIPS mRNA was decreased in genotypes containing four, two or no resistance alleles. The specific activity of the MIPS enzyme in vitro followed the same pattern across genotypes. The IP(3) content in the inoculated roots of genotypes with two, four or six resistance alleles were higher compared to the non-inoculated root. The results suggests that a non-additive effect on transcription and translation of MIPS is established in RIL 23 roots by pyramiding six QTLs for resistance to SDS. A role of MIPS in the partial resistance or response of soybean roots to F. solani infection is suggested. PMID- 12582890 TI - Simple sequence repeat map of the sunflower genome. AB - Several independent molecular genetic linkage maps of varying density and completeness have been constructed for cultivated sunflower ( Helianthus annuus L.). Because of the dearth of sequence and probe-specific DNA markers in the public domain, the various genetic maps of sunflower have not been integrated and a single reference map has not emerged. Moreover, comparisons between maps have been confounded by multiple linkage group nomenclatures and the lack of common DNA markers. The goal of the present research was to construct a dense molecular genetic linkage map for sunflower using simple sequence repeat (SSR) markers. First, 879 SSR markers were developed by identifying 1,093 unique SSR sequences in the DNA sequences of 2,033 clones isolated from genomic DNA libraries enriched for (AC)(n) or (AG)(n) and screening 1,000 SSR primer pairs; 579 of the newly developed SSR markers (65.9% of the total) were polymorphic among four elite inbred lines (RHA280, RHA801, PHA and PHB). The genetic map was constructed using 94 RHA280 x RHA801 F(7) recombinant inbred lines (RILs) and 408 polymorphic SSR markers (462 SSR marker loci segregated in the mapping population). Of the latter, 459 coalesced into 17 linkage groups presumably corresponding to the 17 chromosomes in the haploid sunflower genome ( x = 17). The map was 1,368.3-cM long and had a mean density of 3.1 cM per locus. The SSR markers described herein supply a critical mass of DNA markers for constructing genetic maps of sunflower and create the basis for unifying and cross-referencing the multitude of genetic maps developed for wild and cultivated sunflowers. PMID- 12582891 TI - Analysis on additive effects and additive-by-additive epistatic effects of QTLs for yield traits in a recombinant inbred line population of rice. AB - A linkage map consisting of 158 DNA markers were constructed by using a recombinant inbred line (RIL) population derived from the indica-indica rice cross Zhenshan 97B x Milyang 46. Quantitative trait loci (QTLs) conditioning grain yield and five yield component traits were determined at the one-locus and two-locus levels, and genotype-by-environment (GE) interactions were analyzed. Thirty-one QTLs were detected to have significant additive effects for yield traits, of which 12 also exhibited significant epistatic effects. Sixteen significant additive-by-additive (AA) interactions were detected, of which nine occurred between QTLs with own additive effects (M(ep)QTLs), four occurred between QTLs showing epistatic effects only (epQTLs), and three occurred between M(ep)QTLs and epQTLs. Significant GE interactions were found for six QTLs with additive effects and one AA interaction. Generally, the contributions to the phenotypic variation were higher due to QTL main effects than to epistatic effects. The detection of additive effects and AA effects of a QTL interfered with each other, indicating that the detection of QTLs with main effects, as well as the magnitude and directions of the additive effects, might vary depending on their interactions with other loci. PMID- 12582892 TI - Inheritance of superoxide dismutase ( Sod-1) in a perennial x annual ryegrass cross and its allelic distribution among cultivars. AB - Identifying annual ryegrass contamination in perennial ryegrass seed lots has been of major interest in seed-testing laboratories and for seed regulatory agencies in the USA for many years. This study was conducted to characterize a superoxide dismutase locus ( Sod-1) and determine its potential to distinguish cultivated ryegrass species. The inheritance of Sod-1 was evaluated in a three generation annual x perennial ryegrass mapping population and segregation fitted an expected 1:2:1 ratio for a single locus with two alleles. The molecular form of the Sod-1 locus was determined by H(2)O(2) and KCN inhibitor assays which indicated that the Sod-1, and a second independently segregating Sod-2, locus were both Cu/ZnSod enzymes. The common alleles at the Sod-1 locus were scored in 13 annual and 24 perennial ryegrass cultivars to determine the potential of using this locus for species separation. The Sod-1b allele was homozygous in 98% of perennial ryegrass individuals from 24 cultivars, but those not 100% homozygous for Sod-1b were seed lots with unknown contamination from annual ryegrass. These results indicate that the Sod-1b allele in the homozygous condition is a good indicator of perenniality. All eight annual ryegrass cultivars originating in Europe or Asia had a low frequency of Sod-1b homozygous individuals or none at all. The five cultivars originating in the Western Hemisphere, however, had genotype frequencies for homozygous Sod-1b of up to 56%. The potential of the Sod 1 locus to serve as a test to separate the two growth forms depends on the source of the annual-type contamination. PMID- 12582893 TI - Construction of a BAC library and its application to the identification of simple sequence repeats in peach [ Prunus persica (L.) Batsch]. AB - The Rosaceae contains many economically important crop species, but their genomes are not well characterized, and comparative genetic mapping lags well behind that of other families. To facilitate genome comparisons and gene discovery in the Rosaceae, we have begun the development of genomic resources for peach as the model genome for this family. First, we developed a simplified, cost-effective method for constructing BAC libraries, particularly appropriate for plant species of relatively minor economic importance. Second, we used the library to investigate the abundance and local distribution of simple sequence repeats (SSRs) in peach. Our results indicate that microsatellite loci are locally much more highly abundant than previously estimated, and BAC sequencing results suggest that microsatellite repeats are not randomly distributed within gene containing regions of the peach genome. This makes it relatively easy to identify SSRs in peach by hybridization to BAC clones, and even by random sequencing of BAC clones, not known a priori to contain SSRs. PMID- 12582894 TI - Phylogenetic analysis of Brassiceae based on the nucleotide sequences of the S locus related gene, SLR1. AB - Nucleotide sequences of orthologs of the S-locus related gene, SLR1, in 20 species of Brassicaceae were determined and compared with the previously reported SLR1 sequences of six species. Identities of deduced amino-acid sequences with Brassica oleracea SLR1 ranged from 66.0% to 97.6%, and those with B. oleracea SRK and SLR2 were less than 62% and 55%, respectively. In multiple alignment of deduced amino-acid sequences, the 180-190th amino-acid residues from the initial methionine were highly variable, this variable region corresponding to hypervariable region I of SLG and SRK. A phylogenetic tree based on the deduced amino-acid sequences showed a close relationship of SLR1 orthologs of species in the Brassicinae and Raphaninae. Brassica nigra SLR1 was found to belong to the same clade as Sinapis arvensis and Diplotaxis siifolia, while the sequences of the other Brassica species belonged to another clade together with B. oleracea and Brassica rapa. The phylogenetic tree was similar to previously reported trees constructed using the data of electrophoretic band patterns of chloroplast DNA, though minor differences were found. Based on synonymous substitution rates in SLR1, the diversification time of SLR1 orthologs between species in the Brassicinae was estimated. The evolution and function of SLR1 and the phylogenetic relationship of Brassiceae plants are discussed. PMID- 12582895 TI - Molecular linkage map of allotetraploid cotton ( Gossypium hirsutum L. x Gossypium barbadense L.) with a haploid population. AB - In the present study, a haploid population from the cross of the two cultivated allotetraploid cottons, Gossypium hirsutum L. and Gossypium barbadense L., was developed by means of Vsg, a virescently marked semigamous line of Sea island cotton, and some target haploids were successfully doubled with colchicine. A molecular linkage map was constructed with 58 doubled and haploid plants. Among the total of 624 marker loci (510 SSRs and 114 RAPDs), 489 loci were assembled into 43 linkage groups and covered 3,314.5 centi-Morgans (cM). Using the monosomic and telodisomic genetic stocks, the linkage groups of the present map were associated with chromosomes of the allotetraploid genome, and some of the unassociated groups were connected to corresponding A or D subgenomes. Through the analysis of the assignment of the duplicated SSR loci in the chromosomes or the linkage groups, ten pairs of possible homoeologous chromosome (or linkage group) regions were identified. Among them, the pairs of Chrs. 1 and 15, Chrs. 4 and 22, and Chrs. 10 and 20 had already been determined as homoeologous by classical genetic and cytogenetic research, and the pair of Chrs. 9 and 23 had also been identified by the ISH method of molecular cytogenetics. But, from present research, it was assumed that Chrs. 5 and 18 might be a new pair of homoeologous chromosomes of the allotetraploid cotton genome detected by molecular mapping of the cotton genome. PMID- 12582896 TI - Genetic analysis of floral anthocyanin pigmentation traits in Asiatic hybrid lily using molecular linkage maps. AB - To understand the genetic background of two floral anthocyanin pigmentation traits, anthocyanin pigmentation in the flower tepals and spot formation, in the Asiatic hybrid lily (2n = 24), segregation of the two traits among 96 F(1) plants derived from a cross between commercial cultivars 'Montreux' and 'Connecticut King' were investigated. 'Montreux' has anthocyanin pigmentation in the tepals with many spots, and 'Connecticut King' has flowers with carotenoid pigmentation without spots. The F(1) plants with or without anthocyanin pigment in the tepals segregated with a 1:1 segregation ratio, indicating that a single gene controls anthocyanin pigmentation in the tepals. The number of spots per square centimeter of all tepals showed continuous distribution in the F(1) plants. To map the loci for the two anthocyanin pigmentation traits, molecular linkage maps in the Asiatic hybrid lily were constructed using a double pseudo-testcross strategy, with the same F(1) plants used for phenotypic evaluation, and 212 PCR-based DNA markers. The trait for anthocyanin pigmentation in tepals was used as a trait marker. The map of 'Montreux' comprised 95 markers in 26 linkage groups, and the map of 'Connecticut King' used 119 markers in 24 linkage groups. The total map lengths were 867.5 and 1,114.8 cM, respectively. The trait locus for anthocyanin pigmentation in the tepals was between markers ASR35-180 and P506-40 in linkage group 1 of the 'Montreux' map with a map distance of 1.2 cM and 2.6 cM, respectively. A single-point analysis of quantitative trait loci (QTLs) for tepal spot number identified two putative QTLs in linkage groups 1 and 19 of the 'Connecticut King' map. One putative QTL in linkage group 19 explained 64% of the total phenotypic variation. Because both putative QTLs were mapped on the linkage map of 'Connecticut King' that has no spots, dominant alleles of them might suppress spot formation. PMID- 12582897 TI - Considerations of correlated fertility between genders on genetic diversity: the Pinus densiflora seed orchard as a model. AB - The correlation between 99 clone female and male fertilities in a first generation seed orchard of Pinus densiflora was studied over 6 years. The effective number of the parent ( N(p)) and the variance effective population number [ N(e)((v))] were used to assess the impact of total (Psi(T)), female (psi(f)) and male (psi(m)) fertility variation. A theoretical framework was developed to account for female and male fertility correlations as well as the impact of possible pollen contamination. Total fertility variation was described by the sibling coefficient (Psi(T): the probability that two genes randomly chosen from the gamete gene pool originate from the same parent), which was further subdivided into psi(f) and psi(m). These parameters were compared under various conditions including the total seed harvest, imposing on equal seed harvest among the orchard's clones and two contamination scenarios ( M = 0 and 20%). Fertility variations among females, males and clones were observed within and among years. Sibling coefficients (Psi(T)) were lower, but the effective number of parent ( N(p)) and variance effective population number ( N(e)((v))) were higher in years with moderate female and good male strobilus production. N(p) for female and male reproductive outputs varied from 49 to 82 and from 57 to 93, respectively. N(p) was higher for males than females. When the crop of the 6 years was pooled, N(p) for female, male and the clone were 73, 87 and 85, respectively. The impact of female-male fertility correlation for conditions with no-, positive- and negative-correlations were assessed and their impact on Psi(T), N(p) and N(e)((v)) was also evaluated. It was demonstrated that the practice of equal seed harvesting from every clone, or the mixing of seeds from several years, would substantially improve the genetic diversity and the genetic representation of the seed orchard population when a positive correlation between gender fertilities was observed. The relevance of these results to supplemental mass-pollination was discussed under two cases where equal- and un-equal amounts of pollen from clones were included in the pollen mixes. PMID- 12582898 TI - Conversion of AFLP fragments tightly linked to SCMV resistance genes Scmv1 and Scmv2 into simple PCR-based markers. AB - In a previous study, bulked segregant analysis with amplified fragment length polymorphisms (AFLPs) identified several markers closely linked to the sugarcane mosaic virus resistance genes Scmv1 on chromosome 6 and Scmv2 on chromosome 3. Six AFLP markers (E33M61-2, E33M52, E38M51, E82M57, E84M59 and E93M53) were located on chromosome 3 and two markers (E33M61-1 and E35M62-1) on chromosome 6. Our objective in the present study was to sequence the respective AFLP bands in order to convert these dominant markers into more simple and reliable polymerase chain reaction (PCR)-based sequence-tagged site markers. Six AFLP markers resulted either in complete identical sequences between the six inbreds investigated in this study or revealed single nucleotide polymorphisms within the inbred lines and were, therefore, not converted. One dominant AFLP marker (E35M62 1) was converted into an insertion/deletion (indel) marker and a second AFLP marker (E33M61-2) into a cleaved amplified polymorphic sequence marker. Mapping of both converted PCR-based markers confirmed their localization to the same chromosome region (E33M61-2 on chromosome 3; E35M62-1 on chromosome 6) as the original AFLP markers. Thus, these markers will be useful for marker-assisted selection and facilitate map-based cloning of SCMV resistance genes. PMID- 12582899 TI - From Arabidopsis thaliana to Brassica napus: development of amplified consensus genetic markers (ACGM) for construction of a gene map. AB - The evolution of genomes can be studied by comparing maps of homologous genes which show changes in nucleic acid sequences and chromosome rearrangements. In this study, we developed a set of 32 amplified consensus gene markers (ACGMs) that amplified gene sequences from Arabidopsis thaliana and Brassica napus. Our methodology, based on PCR, facilitated the rapid sequencing of homologous genes from various species of the same phylogenetic family and the detection of intragenic polymorphism. We found that such polymorphism principally concerned intron sequences and we used it to attribute a Brassica oleracea or Brassica rapa origin to the B. napus sequences and to map 43 rapeseed genes. We confirm that the genetic position of homologous genes varied between B. napus and A. thaliana. ACGMs are a useful tool for genome evolution studies and for the further development of single nucleotide polymorphism suitable for use in genetic mapping and genetic diversity analyses. PMID- 12582900 TI - Mapping of qualitative and quantitative phenotypic traits in Rosa using AFLP markers. AB - A segregating population of 91 hybrids issued from a cross between a dihaploid rose, derived from the haploidisation of a modern cultivar, and a diploid species was used to construct linkage maps of the parental genomes. As in other recent genetic studies in Rosa, AFLPs were used as molecular markers. Two segregating qualitative traits, recurrent blooming and double corolla, already known to be inherited as single recessive and dominant genes, respectively, were recorded in the mapping population. A quantitative trait, thorn density of the shoots, was also evaluated in this population. Sixty eight and 108 AFLP markers located on 8 and 6 linkage groups could be analysed in the female and male parent, respectively. The two recorded qualitative phenotypic markers were mapped as well as the quantitative one, after having performed QTL analyses on the parental maps in the latter case. It appears that thorn quantity is controlled by a major and a minor QTL which are located on the same linkage group at 36.5 and 3.2 cM from the single seasonal-blooming gene, respectively. PMID- 12582901 TI - Assessment of peroxidase isozyme marker-based model for cross identifications in hybrids (F(1)) of urdbean [ Vigna mungo (L.) Hepper]. AB - Four hybrids (4 F(1)s) were chosen out of crosses in the urdbean [ Vigna mungo (L.) Hepper, 2n = 22] having contrasting morphological characters. Zymograms for isozyme peroxidase were drawn from the patterns obtained from parents and their respective F(1) hybrids on the basis of relative similarities to parental bands. The selfed or crossed nature of hybrid pods was determined from the zymograms and their analysis. The number of bands and their intensities gave an idea about the extent of crossing in F(1) populations. Genetic identity (I) values were indicative of their selfed nature. Dendrograms were constructed on the basis of genetic identity values to display the relative similarities between the populations. Analysis was based on individual pods to confirm their hybrid or selfed nature. Possible use of this technique for identification of F(1) pods and elimination of selfed pods might be implemented to shorten the breeding operations during crossing. PMID- 12582902 TI - Analysis of genetic relationships among perennial and annual Cicer species growing in Turkey using RAPD markers. AB - Random amplified polymorphic DNA (RAPD) fragments were used to assess genetic relationships among Cicer spp. growing in Turkey. Seven 10-mer primers selected from a 50 random oligonucleotide primer set, depending on their ability to amplify genomic DNA in all species, were used to detect RAPD variation in 43 wild and cultivated accessions representing ten species. These primers yielded 95 reproducible amplification products, 92 of which were polymorphic. Pairwise genetic distances of accessions estimated according to Nei and Li (1979) were used to produce a dendrogram using UPGMA. The dendrogram contained two main clusters, one of which comprised accessions of the four perennial species ( Cicer montbretii, Cicer isauricum, Cicer anatolicum and Cicer incisum) together with the accessions of the three annual species ( Cicer pinnatifidum, Cicer judaicum and Cicer bijugum), and the other cluster included the remaining three annual species ( Cicer echinospermum, Cicer reticulatum and Cicer arietinum). Analysis of RAPD variation showed that C. incisum is the most similar perennial species to annuals, and C. reticulatum is the closest annual species to chickpea. These results generally agree with our allozyme study which was carried out using same Cicer collection and previous studies of relationships among annual species. The results also show that RAPD markers can be used to distinguish Cicer species and to survey genetic variation and relationships among taxonomic units in this genus. PMID- 12582903 TI - Overexpression of the wasabi defensin gene confers enhanced resistance to blast fungus ( Magnaporthe grisea) in transgenic rice. AB - Transgenic rice ( Oryza sativa cv. Sasanishiki) overexpressing the wasabi defensin gene, a plant defensin effective against the rice blast fungus, was generated by Agrobacterium tumefaciens-mediated transformation. Twenty-two T2 homozygous lines harboring the wasabi defensin gene were challenged by the blast fungus. Transformants exhibited resistance to rice blast at various levels. The inheritance of the resistance over generations was investigated. T3 plants derived from two highly blast-resistant T2 lines (WT14-5 and WT43-5) were challenged with the blast fungus using the press-injured spots method. The average size of disease lesions of the transgenic line WT43-5 was reduced to about half of that of non-transgenic plants. The 5-kDa peptide, corresponding to the processed form of the wasabi defensin, was detected in the total protein fraction extracted from the T3 progeny. Transgenic rice plants overproducing wasabi defensin are expected to possess a durable and wide-spectrum resistance (i.e. field resistance) against various rice blast races. PMID- 12582904 TI - AhCMO, regulated by stresses in Atriplex hortensis, can improve drought tolerance in transgenic tobacco. AB - Choline monooxygenase (CMO) catalyzes the committed step of glycine betaine (GlyBet) biosynthesis in many flowering plants. To investigate its effect on various stress tolerances in plant metabolic engineering, we isolated and characterized the CMO gene from Atriplex hortensis, a GlyBet natural accumulator, and introduced it into tobacco to examine the effect of GlyBet on plant drought and salt tolerance, respectively. In A. hortensis, the expression of AhCMO was induced 3-fold in the root and stem, as well as in the leaf, when plants were treated with 400 mM of NaCl, indicating that the acceleration of GlyBet biosynthesis under salt stress was achieved through the whole plant, including organs without chloroplasts. AhCMO transcription was also regulated by drought, ABA and circadian rhythm. Over-expression of AhCMO improved drought tolerance in transgenic tobacco when cultured in medium containing PEG-6000. The transgenic plants also have a better performance under salt stress. PMID- 12582905 TI - Self-fertile cybrids Nicotiana tabacum (+ Hyoscyamus aureus) with a nucleo plastome incompatibility. AB - Cytoplasmic hybrids (cybrids) in a novel inter-generic combination, Nicotiana tabacum (+ Hyoscyamus aureus), were generated by fusion of protoplasts from a plastome tobacco albino mutant (line R100a1) and gamma-irradiated green protoplasts of H. aureus. Cybrids possessed a plastome of H. aureus and a rearranged mitochondrial DNA. The cybrids displayed a syndrome of nucleo-plastome incompatibility expressed as a partial chlorophyll-deficiency of cotyledonary and true leaves at the early stage of vegetative development of plants grown from seeds in soil. During later development, the plants restored a normal green coloration. This character is phenotypically indistinguishable from the same syndrome in previously generated cybrids N. tabacum (+ H. nigrum). In contrast to the cybrids N. tabacum (+ H. nigrum), cybrids N. tabacum (+ H. aureus) were self fertile, and did not manifest other features that were interpreted as nucleo mitochondrial incompatibilities in N. tabacum (+ H. nigrum) plants. Therefore, the cybrids N. tabacum (+ H. aureus) present a self-propagating system of Nicotiana (+ Hyoscyamus) nucleo-plastome incompatibility in its pure form. PMID- 12582906 TI - Genomic relationships between Medicago murex Willd. and Medicago lesinsii E. Small. investigated by in situ hybridization. AB - Medicago murex Willd. is an annual species (2n = 14) widespread in the wild and of remarkable interest for pastures in regions with a mediterranean climate. It is considered closely related to Medicago lesinsii E. Small (2n = 16) but, up to now, there is no evidence demonstrating their genetic affinity. This research was undertaken to investigate the genomic relationships between M. murex and M. lesinsii by using genomic in situ hybridization (GISH). In this study GISH experiments were performed using both species as sources of chromosomes and genomic probes. To better evaluate the results of the hybridization, the labelled DNA of each species was hybridized to chromosomes of the same species and to chromosomes of the diploid Medicago littoralis (2n = 16). Strong hybridization signals were found on chromosomes of M. murex and M. lesinsii after GISH. Differences in the hybridization strength were not observed when slides from interspecific hybridization were compared with the control preparations. These results suggest that consistent divergences of the DNA sequences did not occur after the separation of the two species. Instead very reduced cross hybridization was found on chromosome spreads of M. littoralis hybridized with the DNA of M. lesinsii or M. murex. The distribution of the ribosomal genes (rDNA) investigated by fluorescent in situ hybridization (FISH) appeared similar in both M. murex and M. lesinsii. The GISH technique may be a valuable approach to obtain information on evolution of the 2n = 14 species and on the origin of the polyploids Medicago rugosa (2n = 30) and Medicago scutellata (2n = 30). The first attempt to investigate the genomic composition of M. scutellata using a genomic probe is reported in this paper. PMID- 12582907 TI - Intergeneric hybrids between Brassica napus and Orychophragmus violaceus containing traits of agronomic importance for oilseed rape breeding. AB - Protoplast fusions between Brassica napus and Orychophragmus violaceus for transfer of valuable traits to oilseed rape resulted in 257 somatic hybrid plants. Hybridity was confirmed by morphological, cytological and molecular means. Symmetric fusions gave rise to 131 plants. Fifty eight of these plants had an intermediate morphology and contained nuclear DNA corresponding to the sum of the parental species. All 131 plants were sterile with no pollen grains observed upon flowering. Another 126 plants were derived from asymmetric fusions in which protoplasts of the donor parent O. violaceus were irradiated by 100 or 200-Gy X rays prior to fusion. Morphologically these plants showed a larger variation compared to the plants regenerated from symmetric fusion experiments. In contrast to plants obtained from symmetric fusions, fertile hybrids were recovered among regenerants from the asymmetric fusions. Twenty four of these plants released viable pollen grains and 14 of the determined 17 plants set seeds after either selfing or backcrossing to B. napus. Fourteen male-sterile plants were identified with female fertility. This observed male sterility most-likely originated from alloplasmic recombination and would be of great potential for the development of a new cytoplasmic male sterility system. The fatty acid composition of the fertile hybrids and their progenies showed a biased distribution towards the B. napus parent, which has a high erucic acid-content type. However, increased levels of palmitic and linoleic acids compared to B. napus were found in subsequent generations, as well as a reduced level of erucic acid. PMID- 12582908 TI - Sequence variation in the gene encoding the 10-kDa prolamin in Oryza (Poaceae). I. Phylogenetic Implications. AB - Oryza L. (Poaceae) contains approximately 20 wild and two domesticated species and nine genomes. Major disagreements exist on its systematics and genome evolution. Sequence polymorphism in the gene that encodes the 10-kDa prolamin polypeptide (a seed storage protein) was used to determine phylogenetic relationships and evaluate current systematics for 19 Oryza species. This gene in Oryza is approximately 402-bp long, and includes a 72-bp signal peptide region. A strict consensus tree shows Oryza brachyantha (FF) as the most basal species, followed by a polytomy of three clades that can be delineated based on genome composition: (1) the GG clade: Oryza granulata and Oryza meyeriana, (2) the EE clade: Oryza australiensis, and (3) the ABCD clade: the remaining Oryza species. Two subclades within the ABCD clade emerge, one containing species with the AA genome, the other with components of the BC and D genomes. Members of the AA subclade form a polytomy and were delineated by a single 3-base deletion. The African species Oryza punctata (BB) and the South American-endemic CCDD genome species form a strong lineage, pointing to a close genetic affinity of O. punctata to the missing DD genome donor. The strong association between the CC and BBCC species implies convergence at the gene level. The study supports the following sectional units of Oryza: Section Oryza (Series sativae and officinaliae), Section australiensis, Section Granulata, Section Brachyantha. PMID- 12582909 TI - Integration of sequence tagged microsatellite sites to the chickpea genetic map. AB - Fifty sequence-tagged microsatellite site (STMS) markers and a resistant gene analog (RGA) locus were integrated into a chickpea ( Cicer arietinum L., 2n = 2 x = 16 chromosomes) genetic map that was previously constructed using 142 F(6) derived recombinant inbred lines (RILs) from a cross of C. arietinum x Cicer reticulatum Lad. The map covers 1,174.5 cM with an average distance of 7.0 cM between markers in nine linkage groups (LGs). Nine markers including the RGA showed distorted segregation ( P < 0.05). The majority of the newly integrated markers were mapped to marker-dense regions of the LGs. Six co-dominant STMS markers were integrated into two previously reported major quantitative trait loci (QTLs) conferring resistance to Ascochyta blight caused by Ascochyta rabiei (Pass.) Labr. Using common STMS markers as anchors, three maps developed from different mapping populations were joined, and genes for resistance to Ascochyta blight, Fusarium wilt (caused by Fusarium oxysporum Schlechtend.: Fr. f. sp. ciceris), and for agronomically important traits were located on the combined linkage map. The integration of co-dominant STMS markers improves the map of chickpea and makes it possible to consider additional fine mapping of the genome and also map-based cloning of important disease resistance genes. PMID- 12582910 TI - Recessive resistance genes against potyviruses are localized in colinear genomic regions of the tomato ( Lycopersicon spp.) and pepper ( Capsicum spp.) genomes. AB - Resistance against both Potato virus Y (PVY) and Tobacco etch virus (TEV) was identified in the wild tomato relative Lycopersicon hirsutum PI247087. Analysis of the segregation ratio in F(2)/F(3) and BC(1) interspecific progenies indicated that a single recessive gene, or two very tightly linked recessive loci, are involved in resistance to both potyviruses. This locus was named pot-1. Using amplified fragment length polymorphism markers and a set of L. hirsutum introgression lines, pot-1 was mapped to the short arm of tomato chromosome 3, in the vicinity of the recessive py-1 locus for resistance to corky root rot. Because of the occurrence of phenotypically similar genes in pepper ( Capsicum spp.), the comparative genetics of resistance to potyviruses between tomato and pepper was investigated. Unlike most of the comparative genetic studies on resistance genes, pot-1 was tightly flanked by the same restriction fragment length polymorphism (RFLP) markers than the pvr2/pvr5 locus for resistance to PVY and TEV from pepper. These results may indicate that recessive resistance genes against potyviruses evolve less rapidly than the majority of the dominant genes cloned so far, and consequently may belong to a different family of resistance genes. PMID- 12582911 TI - Ty1-copia retrotransposon-based S-SAP (sequence-specific amplified polymorphism) for genetic analysis of sweetpotato. AB - RNaseH-LTR regions of the Ty1-copia retrotransposon were amplified and cloned from the sweetpotato genome using RNaseH gene-specific degenerate primers and restriction site-specific adaptor primers. Ninety clones out of the 240 sequenced were identified with a variable degree of homology to the Ty1-copia RNaseH gene. Three ( Str6, Str85, Str187) of the 90 had characteristic RNaseH-gene, stop codon, polypurine track and putative 3' LTR sequence elements. Analysis of nine selected genotypes representing Africa, South and Central America, as well as Papua New Guinea, by the established S-SAP technique revealed that the majority of the Ty1-copia transposon insertions were unique (33 to 64%) and only few common bands were detected. Analysis of 177 East African varieties further supported this finding and showed that most of the copia retrotransposon locations were represented only by some genotypes. Considering that sweetpotato has been present in the East African region for only about 500 years, and the number of genotypes introduced was possibly limited, a surprisingly high level of genetic variability of the transposon insertion sites was detected. These findings may indicate the putative activity of the retrotransposon in sweetpotato in the recent past. Comparison of the copia retrotransposon insertion-based S-SAP method to AFLP and RAPD showed that the majority of the markers were more polymorphic (97-99%) in the case of S-SAP in comparison to AFLP (70-90%) and RAPD (88%). Thus demonstrating the transposon-based molecular marker system was very efficient for genotyping. PMID- 12582912 TI - Expression of wild-type pinB sequence in transgenic wheat complements a hard phenotype. AB - Wheat grain hardness is a major factor in the wheat end-product quality. Grain hardness in wheat affects such parameters as milling yield, starch damage and baking properties. A single locus determines whether wheat is hard or soft textured. This locus, termed Hardness ( Ha), resides on the short arm of chromosome 5D. Sequence alterations in the tryptophan-rich proteins puroindoline a and b (PINA and PINB) are inseparably linked to hard textured grain, but their role in endosperm texture has been controversial. Here, we show that the pinB-D1b alteration, common in hard textured wheats, can be complemented by the expression of wild-type pinB-D1a in transformed plants. Transgenic wheat seeds expressing wild-type pinB were soft in phenotype, having greatly increased friabilin levels, and greatly decreased kernel hardness and damaged starch. These results indicate that the pinB-D1b alteration is most likely the causative Ha mutation in the majority of hard wheats. PMID- 12582913 TI - Transgene behaviour across two generations in a large random population of transgenic rice plants produced by particle bombardment. AB - The relationship between transgene copy number, rearrangement levels, inheritance patterns, expression levels, transgene stability and plant fertility was analysed in a random population of 95 independently transformed rice plant lines. This analysis has been conducted for both the selectable marker gene ( aphIV) and the unselected reporter gene ( gusA), in the presence or absence of flanking Matrix Attachment Regions (MARs) in order to develop a better understanding of transgene behaviour in a population of transgenic rice plants created by particle bombardment. In the first generation (T(0)), all the independently transformed plant lines contained and expressed the aphIV gene conferring resistance to hygromycin, but only 87% of the lines were co-transformed with the unselected gusA marker gene. Both transgenes seemed to be expressed independently. Most lines exhibited complex transgene rearrangements as well as an intact transgene expression unit for both aphIV and gusA transgenes. Transgene copy number was proportional to the quantity of DNA used during bombardment. In T(0) plants, high gusA copy number significantly decreased GUS expression levels but there was no correlation between expression level and transgene copy number across the entire population of lines. Four main factors impaired transgene expression in primary transgenic plants (T(0)) and their progeny (T(1)): (1) absence of transgene expression in T(0) plants (41% of lines), (2) sterility of T(0) plants (28% of lines), (3) non-transmission of intact transgenes to some or all progenies (at least 14% of lines), and (4) silencing of transgene expression in progeny plants (10% of lines). Transgene stability was significantly related to differences in transgene structure and expression levels. The presence of Rb7 MARs flanking the gusA expression unit had no effect on plant fertility or non-transmission of transgenes, but provided copy number-dependent expression of the transgene and improved expression levels and stability over two generations. Overall, only 7% of the plant lines without MARs and 17% of the lines with MARs initially generated, exhibited stable transgene expression over two generations. PMID- 12582914 TI - Substitution mapping of Pup1: a major QTL increasing phosphorus uptake of rice from a phosphorus-deficient soil. AB - A major QTL for P uptake had previously been mapped to a 13-cM marker interval on the long arm of chromosome 12. To map that major QTL with higher precision and certainty, a secondary mapping population was developed by backcrossing a near isogenic line containing the QTL from the donor parent to the recurrent parent of low P uptake. Two different mapping strategies have been followed in this study. A conventional QTL mapping approach was based on individual F(2) RFLP data and the phenotypic evaluation of family means in the F(3). The second strategy employed a substitution-mapping approach. Phenotypic and marker data were obtained for 160 F(3) individuals of six highly informative families that differed in the size of donor chromosomal segments in the region of the putative QTL. QTL mapping showed that close to 80% of the variation between families was due to a single QTL, hereafter referred to as Pup1 (Phosphorus uptake 1). Pup1 was placed in a 3-cM interval flanked by markers S14025 and S13126, which is within 1 cM of the position identified in the original QTL mapping experiment. Other chromosomal regions and epistatic effects were not significant. Substitution mapping revealed that Pup1 co-segregated with marker S13126 and that the flanking markers, S14025 and S13752, were outside the interval containing Pup1. The two mapping strategies therefore yielded almost identical results and, in combining the advantages of both, Pup1 could be mapped with high certainty. The QTL mapping appoach showed that the phenotypic variation between families was due to only one QTL without any additional epistacic interactions, whereas the advantage of substitution mapping was to place clearly defined borders around the QTL. PMID- 12582915 TI - Microsatellites in starch-synthesizing genes in relation to starch physicochemical properties in waxy rice ( Oryza sativa L.). AB - Rice starch is composed of amylose and amylopectin. Amylose content, an important determinant of rice starch quality, is primarily controlled by the waxy gene, encoding granule-bound starch synthase (GBSS). The starch branching enzyme (SBE) and soluble starch synthase (SSS) play major roles in the synthesis of amylopectin. Microsatellite polymorphisms in the three genes, the wx gene encoding granule-bound starch synthase I, the SBE gene encoding starch branching enzyme I and the SSS gene encoding soluble starch synthase I, were studied for 56 accessions of waxy rice ( Oryza sativa L.). Four (CT)(n) microsatellite alleles, (CT)(16), (CT)(17), (CT)(18) and (CT)(19), at the wx locus were detected in this set of waxy rice, of which (CT)(17) was the most frequent. Three (CT)(n) microsatellite allele classes were found at the SBE locus, (CT)(8) or (CT)(10) together with an insertion sequence of CTCTCGGGCGA, and (CT)(8) alone without the insertion. There were multiple microsatellites clustered at the SSS locus. However, these alleles can also be grouped into three classes, i.e. the allele class SSS-A = (AC)(2) em leader TCC(TC)(11) em leader (TC)(5)C(ACC)(11), the allele class SSS-B = (AC)(3) em leader TCT(TC)(6) em leader (TC)(4)C(ACC)(9), and the allele class SSS-C = (AC)(3) em leader TCT(TC)(6) em leader (TC)(4)C(ACC)(8). The analyses of starch physicochemical properties among different microsatellite genotypes indicated that the waxy rice group with the (CT)(19) allele, the SBE-A allele and the SSS-B allele was quite different from other groups. Nine out of 15 accessions with a high gelatinization temperature (GT) belonged to the wx (CT)(19) group, all of them belonged to the SBE-A group and 13 of them belonged to the SSS-B group. These microsatellites might be useful in marker-assisted breeding for the improvement of rice grain quality. PMID- 12582916 TI - Genetic basis of hybrid breakdown in a Japonica/Indica cross of rice, Oryza sativa L. AB - Reproductive barriers often arise in hybrid progeny between two varietal groups of Asian cultivated rice ( Oryza sativa L.), Japonica and Indica. Hybrid breakdown showing poor growth habit, and complete sterility was found in the backcrossed progeny derived from a cross between a Japonica variety, Asominori, and an Indica variety, IR24. We employed RFLP analysis in the segregating population to study the genetic basis underlying hybrid breakdown. It was found that the hybrid breakdown is caused by a set of two nuclear genes, which were symbolized as hwe1 and hwe2. The parental varieties, Asominori and IR24, carry hwe1(+) hwe1(+) hwe2hwe2 and hwe1hwe1hwe2(+) hwe2(+) genotypes, respectively, whereas the progenies that showed a weakness performance carry the double recessive genotype ( hwe1hwe1hwe2hwe2). Abnormality was not observed in the progenies that carry the other genotypes, indicating that a single dominant allele at either locus is necessary for normal growth. Based on linkage analysis with RFLP markers, the hwe1 locus was located between RFLP markers R1869 and S1437 on chromosome 12 and the hwe2 locus was located between R3192 and C1211 on chromosome 1. The genetic basis was reconfirmed using near-isogenic lines carrying the genes with reciprocal genetic backgrounds. The present study provides clear evidence, viewed by previous workers, that hybrid breakdown is attributed to complementary genes from both parents. PMID- 12582917 TI - Characterization of RFLP probe sequences for gene discovery and SSR development in Sorghum bicolor (L.) Moench. AB - In this study, we collected and analyzed DNA sequence data for 789 previously mapped RFLP probes from Sorghum bicolor (L.) Moench. DNA sequences, comprising 894 non-redundant contigs and end sequences, were searched against three GenBank databases, nucleotide (nt), protein (nr) and EST (dbEST), using BLAST algorithms. Matching ESTs were also searched against nt and nr. Translated DNA sequences were then searched against the conserved domain database (CDD) to determine if functional domains/motifs were congruent with the proteins identified in previous searches. More than half (500/894 or 56%) of the query sequences had significant matches in at least one of the GenBank searches. Overall, proteins identified for 148 sequences (17%) were consistent among all searches, of which 66 sequences (7%) contained congruent coding domains. The RFLP probe sequences were also evaluated for the presence of simple sequence repeats (SSRs) and 60 SSRs were developed and assayed in an array of sorghum germplasm comprising inbreds, landraces and wild relatives. Overall, these SSR loci had lower levels of polymorphism ( D = 0.46, averaged over 51 polymorphic loci) compared with sorghum SSRs that were isolated by library hybridization screens ( D = 0.69, averaged over 38 polymorphic loci). This result was probably due to the relatively small proportion of di-nucleotide repeat-containing markers (42% of the total SSR loci) obtained from the DNA sequence data. These di-nucleotide markers also contained shorter repeat motifs than those isolated from genomic libraries. Based on BLAST results, 24 SSRs (40%) were located within, or near, previously annotated or hypothetical genes. We determined the location of 19 of these SSRs relative to putative coding regions. In general, SSRs located in coding regions were less polymorphic ( D = 0.07, averaged over three loci) than those from gene flanking regions, UTRs and introns ( D = 0.49, averaged over 16 loci). The sequence information and SSR loci generated through this study will be valuable for application to sorghum genetics and improvement, including gene discovery, marker assisted selection, diversity and pedigree analyses, comparative mapping and evolutionary genetic studies. PMID- 12582918 TI - Mapping of quantitative trait loci determining agronomic important characters in hexaploid wheat ( Triticum aestivum L.). AB - A set of 114 recombinant inbred lines of the 'International Triticeae Mapping Initiative' mapping population was grown during the seasons 1997, 1998, 1999 and 2000 under several environments. Twenty morphological (glume colour, awn colour, waxiness, leaf erectness, peduncle length), agronomical (ear emergence time, flowering time, grain filling time, ear length, plant height, lodging, grain number, thousand-grain-weight, grain weight per ear, grain protein content, winter hardiness) and disease resistance (powdery mildew, yellow rust, leaf rust, fusarium) traits were studied. Not all traits were scored in each experiment. In total 210 QTLs with a LOD threshold of >2.0 (minor QTLs) were detected of which 64 reached a LOD score of >3.0 (major QTLs). Often QTLs were detected in comparable positions in different experiments. Homologous and homoeologous relationships of the detected QTLs, and already described major genes or QTLs determining the same traits in wheat or other Triticeae members, are discussed. PMID- 12582919 TI - Expression and inheritance of the wheat Glu-1DX5 gene in transgenic maize. AB - We have produced transgenic maize plants containing a wheat Glu-1Dx5 gene encoding the high-molecular-weight glutenin subunit 1Dx5. Analysis by SDS-PAGE showed that a protein similar in size to the wheat 1Dx5 subunit accumulates in the endosperm of transgenic maize from four independent transformation events. This protein reacts with a monoclonal antibody specific to the wheat 1Dx5 subunit and was not detected in nontransgenic controls or in pollen, anthers, leaves or embryos of plants grown from seeds expressing this protein in endosperm. Genomic Southern-blot analysis is consistent with results from SDS-PAGE and indicates that the transgene integration sites are complex and are different in the four events studied. Using the presence of this protein as a phenotypic marker, we studied the inheritance of this gene through three sexual generations. Reciprocal crosses with nontransgenic plants and self-pollinations were performed, and the resulting kernels were analyzed for the presence of the 1Dx5 subunit. These data, together with PCR analysis for the transgene, suggest that the transgene is inefficiently transmitted through pollen in all four events. PMID- 12582920 TI - Preliminary genetic linkage map of Miscanthus sinensis with RAPD markers. AB - We have used an "offspring cross" mapping strategy in combination with the random amplified polymorphic DNA (RAPD) assay to construct the first genetic map of the species Miscanthus sinensis (2 n = 2 x = 38). This map is based on an outbred population of 89 individuals resulting from the cross between two genotypes from a previously designed cross. Consequently, both parents are fullsibs. The same proportion of bi-parental markers (heterozygotic in both parents) and pseudo testcross markers (heterozygotic in one parent and null in the other), mono parental markers, have been obtained. A total of 383 RAPD markers were analysed within the 89 F1 plants. Out of these markers, 257 were mapped into 28 linkage groups which spanned a total map length of around 1,074.5 cM with an average density of 4.2 cM per marker. Out of 257 mapped markers, 62 were inherited from F1.1 (P1), 63 from F1.7 (P7) and 132 were bi-parental markers. The contribution to the map was equal from both parents. This map provides a useful tool for genetic analyses of agronomically interesting characters in M. sinensis such as flowering, yield, plant height, stem diameter and mineral constitution. The offspring cross mapping strategy is proposed to obtain a higher efficiency in developing integrated maps including both parents. PMID- 12582921 TI - The origin of intermediate species of the genus Sorbus. AB - The genus Sorbus in Europe contains five diploid species, Sorbus aria, Sorbus aucuparia, Sorbus torminalis, Sorbus chamaemespilus and Sorbus domestica, classified into five different subgenera. The subgenus Aria (or the S. aria aggregate) contains apomictic triploid and tetraploid species. Within the genus there are, in addition, a number of species, morphologically intermediate between plants of the four main subgenera, which are considered to reproduce partly, or exclusively, by apomixis. These are believed to have originated by hybridisation between species in the S. aria aggregate and a species from another subgenus (either S. aucuparia, S. torminalis or S. chamaemespilus). We have used restriction fragment length polymorphism (RFLP) analysis on a total of 178 Sorbus accessions to test this model. The genome relationships of the different groups have been assessed, and the hybrid nature of the major intermediate groups is unequivocally demonstrated. Polyploid species in the S. aria aggregate show genetic variation, indicating the possibility of multiple origins and/or facultative apomictic breeding behaviour. A major finding, confirmed by microsatellite analysis, is that the 'intermediate' species S. intermedia is shown to have genomes from S aria, S aucuparia and S torminalis. Polymorphic mitochondrial DNA markers were used to determine the direction of the crosses that gave rise to new 'hybrid species'; in the majority of cases the pollen was provided by the parent from the S aria aggregate. PMID- 12582922 TI - Analysis of genetic effects of major genes and polygenes on quantitative traits. II. Genetic models for seed traits of crops. AB - Genetic models for quantitative seed traits with effects of several major genes and polygenes, as well as their GE interaction, were proposed. Mixed linear model approaches were suggested for analyzing the genetic models. Monte Carlo simulations were conducted to evaluate unbiasedness and efficiency for estimating fixed effects and variance components of the embryo and the endosperm models, including effects of a major gene from an unbalanced modified diallel mating design with nine parents, respectively. Simulation results showed that estimates of generalized least squares (GLS) were unbiased and efficient, while those of ordinary least squares (OLS) were almost as good as GLS. Minimum norm quadratic unbiased estimation (MINQUE) could obtain unbiased estimates of the variance components. It was also suggested that precision of MINQUE estimation would be improved with augmentation of experimental size. Data from a modified diallel design in upland cotton ( Gossypium hirsutum L.) were used as a worked example to illustrate the parameter estimation. PMID- 12582923 TI - Cloning and mapping of genes involved in wheat-leaf rust interaction through gene expression analysis using chromosome-deleted near-isogenic wheat lines. AB - Molecular markers on wheat chromosome 6BL were isolated using mRNA differential display. Two wheat isolines inoculated with Puccinia recondita were analysed: Sinvalocho MA line carrying the Lr3 gene for leaf rust resistance on distal chromosome 6BL, and a rust-susceptible derivative of the Sinvalocho MA line with a deletion at the distal end of chromosome 6BL. Comparison of mRNA fingerprinting profiles, obtained from control and rust-inoculated plants, let to the isolation of 34 differentially displayed cDNAs. All these genes, except TaRr16, were up regulated in the rust-inoculated resistant line. TaRr16 has constitutive expression in the rust-resistant line while no expression was detected in the rust-susceptible line. A number of those cDNAs revealed homology to genes previously identified in other plant-pathogen interactions. Two out of the 34 cDNAs, mapped in the distal part of chromosome 6BL and TaRr16, was genetically linked to the Lr3 gene. DNA sequence differences and differential expression between non-allelic copies of TaRr16, are also reported. PMID- 12582924 TI - Field evaluation and agronomic performance of transgenic wheat. AB - Seven transgenic lines of wheat have been evaluated under field conditions during 2 agonomic years. Four lines contained the transgenes for beta-glucuronidase ( uidA), herbicide resistance ( bar) and for one high-molecular-weight (HMW) subunit, and three lines contained only one transgene for one HMW glutenin subunit and no marker genes. Agronomic traits and yield components were studied in transgenic lines and compared with the non-transgenic parent and null segregant lines. Although phenotypic differences for many traits have been found, only heading date and the number of spikelets per spike showed clear genotypic differences for both field trials. All transgenic lines had a longer heading date than parent lines whereas the number of spikelets per spike in transgenic lines was around that for L88-31 and higher for L88-6 than the corresponding parent lines. No differences were found between lines constitutively expressing the uidA and bar genes from those which only expressed the HMW genes. We conclude that differences between transgenic lines and their parents are small, and could be eliminated by backcrossing transgenic lines with their parents and selecting for the wanted genotype. PMID- 12582925 TI - Comparative genetic analysis of quantitative traits in sunflower ( Helianthus annuus L.) 1. QTL involved in resistance to Sclerotinia sclerotiorum and Diaporthe helianthi. AB - Sclerotinia sclerotiorum and Diaporthe helianthi are important pathogens of sunflower ( Helianthus annuus L.). Two hundred and twenty F2-F3 families were developed from an intraspecific cross between two inbred sunflower lines XRQ and PSC8. Using this segregating population a genetic map of 19 linkage groups with 290 molecular markers covering 2,318 cM was constructed. Disease resistances were measured in field experiments during 3 years (1998, 1999 and 2000) for phomopsis and 2 years for S. sclerotiorum (1997 and 1999). QTL were detected using the interval mapping method at a LOD threshold of 3. A total of 15 QTL for each pathogen resistance were detected across several linkage groups, confirming the polygenic nature of the resistances. These QTL explained from 7 to 41% of the phenotypic variability. The QTL for phomopsis resistance, in the 3 years of tests, mapped in the same region, and this was also true for some forms of S. sclerotiorum resistance in the 2 years of tests. On linkage group 8, QTL affecting resistance to both S. sclerotiorum and D. helianthi mycelium extension on leaves colocalised, suggesting a common component in the mechanism of resistance for these two pathogens. The colocalisation of QTL and breeding for resistance to S. sclerotiorum and to D. helianthi by pyramiding QTL in sunflower are discussed. PMID- 12582926 TI - Introgression of self-compatibility from Coffea heterocalyx to the cultivated species Coffea canephora. AB - Self-compatibility segregation was assessed in two successive backcross progenies originating from an interspecific cross between Coffea canephora (self incompatible) and Coffea heterocalyx (self-compatible). After self- and cross pollination, pollen tube behaviour in styles was observed under ultraviolet fluorescence microscopy and fruit-set was determined at harvesting time. Segregation ratios in the two progenies were consistent with monofactorial control of self-compatibility. Self-compatible plants exhibited higher fruit-set than self-incompatible ones in open-pollination conditions. Segregation of AFLP markers was scored in the first backcross progeny. By molecular linkage analysis, the S locus could be mapped to a short linkage group. PMID- 12582927 TI - AFLP maps of Petunia hybrida: building maps when markers cluster. AB - AFLP mapping in Petunia hybrida was undertaken with the intention of building a high-density genetic map suitable for applications such as map-based gene cloning. In total five maps were constructed from two mapping populations, with placement of more than 800 markers. Despite the large number of markers the resulting map is roughly ten-fold smaller than those of other plant species, including the closely related tomato. Low levels of recombination are reflected in clusters of tightly linked markers, both AFLPs and RFLPs, in all the maps. Clustering patterns vary between mapping populations, however, such that loci tightly linked in one population may be separable in another. Combined with earlier reports of aberrant meiotic pairing and recombination, our results suggest that, for species like petunia, map-based cloning may be more complex than in model species such as arabidopsis and tomato. PMID- 12582928 TI - Identification of molecular markers associated with leptine in reciprocal backcross families of diploid potato. AB - Solanum phureja clone 1-3 and S. chacoense clone 80-1 have a zero and high leptine content in their foliage, respectively. An F(1) hybrid (CP2) was intermediate for the trait, but self-incompatible. Two reciprocal backcross families, PBCp ( phu 1-3 x CP2) and PBCc (CP2 x phu 1-3), and a family of monoploids derived by anther culture of CP2, were characterized for leptine as the aglycon, acetylleptinidine (ALD), content in leaves by gas chromatography. ALD was present in 43 of 87 genotypes in the PBCp backcross, implying simple genetic control by a dominant gene. However, the ALD levels were low compared to CP2. In the PBCc backcross, only 7 of 42 genotypes expressed ALD at a level generally higher than in PBCp. This ratio was significantly different from the 1:1 segregation observed in the reciprocal backcross and suggests a cytoplasmic influence. ALD levels in the CP2 monoploids ranged from 0 to 8,968 &mgr;g.g(-1) of dry weight (dw) with 18 individuals expressing ALD and five with 0 ALD content. Ten high (mean ALD = 546 &mgr;g.g(-1) of dw) and ten low (mean ALD = 0) individual plants within PBCp and seven high (mean ALD = 3,037 &mgr;g.g(-1) of dw) and eight low (mean ALD = 0) individual plants within PBCc were used for bulk segregant analysis (BSA) using 214 RAPD (randomly amplified polymorphic DNA) primers. Three RAPD primers (OPQ-2, OPT-16 and OPT-20) amplified bands exclusively in bulks containing DNA mixes of high ALD producers in both PBCp and PBCc populations. These results suggest that these markers were associated in coupling to ALD content. ANOVAs for ALD content verified association between the markers and the trait. A CAPS (cleaved amplified polymorphic sequence) marker, GP82A, was also significantly associated with ALD production in both the monoploid and the PBCp populations. None of the RAPD markers was associated to ALD in the monoploids but one was associated in repulsion. The monoploid data indicate the likelihood of a recessive gene(s) that controls leptine production, but the backcross data indicate the action of modifying loci. PMID- 12582929 TI - Construction and testing of a microsatellite database containing more than 500 tomato varieties. AB - The aim of this study was to evaluate the suitability of sequence tagged microsatellite site (STMS) markers for varietal identification and discrimination in tomato. For this purpose, a set of 20 STMS primer pairs was used to construct a database containing the molecular description of the most common varieties (>500) of tomato grown in Europe. The database was built and tested by a consortium of five European laboratories each using a different STMS detection system. In this way, it could be demonstrated that the STMS markers and database were suitable for use in network activities where a common database is being established on a continuing basis with data from different laboratories.Microsatellite polymorphism in tomato was found to be relatively low. The number of alleles per locus ranged from 2 to 8 with an average of 4.7 alleles per locus. Nevertheless, more than 90% of the varieties had different microsatellite profiles. A "blind testing" exercise showed that in general, identification of unknown samples (or detecting the most similar variety) with the 20 markers and the database was relatively easy for homogeneous varieties but less certain with heterogeneous varieties when using pools of 6 individuals. PMID- 12582930 TI - Genetic dissection of a modern sugarcane cultivar ( Saccharum spp.).II. Detection of QTLs for yield components. AB - The genetics of current sugarcane cultivars ( Saccharum spp.) is outstandingly complex, due to a high ploidy level and an interspecific origin which leads to the presence of numerous chromosomes belonging to two ancestral genomes. In order to analyse the inheritance of quantitative traits, we have undertaken an extensive Quantitative Trait Allele (QTA) mapping study based on a population of 295 progenies derived from the selfing of cultivar R570, using about 1,000 AFLP markers scattered on about half of the genome. The population was evaluated in a replicated trial for four basic yield components, plant height, stalk number, stalk diameter and brix, in two successive crop-cycles. Forty putative QTAs were found for the four traits at P = 5 x 10(-3), of which five appeared in both years. Their individual size ranged between 3 and 7% of the whole variation. The stability across years was improved when limiting threshold stringency. All these results depict the presence in the genome of numerous QTAs, with little effects, fluctuating slightly across cycles, on the verge to being perceptible given the experimental resolution. Epistatic interactions were also explored and 41 independent di-genic interactions were found at P = (5 x 10(-3))(2). Altogether the putative genetic factors revealed here explain from 30 to 55% of the total phenotypic variance depending on the trait. The tentative assignment of some QTAs to the ancestral genomes showed a small majority of contributions as expected from the ancestral phenotypes. This is the first extensive QTL mapping study performed in cultivated sugarcane. PMID- 12582931 TI - "Perfect" markers for the Rht-B1b and Rht-D1b dwarfing genes in wheat. AB - PCR-based markers were developed to detect the point mutations responsible for the two major semi-dwarfing genes Rht-B1b ( Rht1) and Rht-D1b ( Rht2) in wheat. These markers were validated by testing 19 wheat varieties of known Rht genotype. They included Rht-B1b and Rht-D1b dwarfs, double-mutant varieties and tall wheats. These were correctly genotyped with the Rht-B1b and Rht-D1b-specific primers, as well as markers specific for the tall alleles Rht-B1a and Rht-D1a. Using a family of doubled-haploid lines segregating for Rht-B1b and Rht-D1b, the markers were mapped to the expected homoeologous regions of chromosomes 4B and 4D, respectively. Both markers were strongly correlated with a reduction in height, accounting for 23% ( Rht-B1b) and 44% ( Rht-D1b) of the phenotypic variance in the population. These markers will have utility in marker-assisted selection of the Rht-B1b and Rht-D1b genes in wheat breeding programs. PMID- 12582932 TI - Mapping of quantitative trait loci based on growth models. AB - An approach called growth model-based mapping (GMM) of quantitative trait loci (QTLs) is proposed in this paper. The principle of the approach is to fit the growth curve of each individual or line with a theoretical or empirical growth model at first and then map QTLs based on the estimated growth parameters with the method of multiple-trait composite interval mapping. In comparison with previously proposed approaches of QTL mapping based on growth data, GMM has several advantages: (1) it can greatly reduce the amount of phenotypic data for QTL analysis and thus alleviate the burden of computation, particularly when permutation tests or simulation are performed to estimate significance thresholds; (2) it can efficiently analyze unbalanced phenotype data because both balanced and unbalanced data can be used for fitting growth models; and (3) it may potentially help us to better understand the genetic basis of quantitative trait development because the parameters in a theoretical growth model may often have clear biological meanings. A practical example of rice leaf-age development is presented to demonstrate the utility of GMM. PMID- 12582933 TI - Association of RFLP markers and biomass heterosis in trigenomic hybrids of oilseed rape ( Brassica napus x B. campestris). AB - Laboratory screening with DNA-based markers and field measurements of biomass production were carried out on each of the 120 trigenomic hybrids, obtained by interspecific hybridization between Brassica napus (AACC) and Brassica campestris (A'A'). The objective of this study was to elucidate the relationship between molecular markers and biomass heterosis of the interspecific hybrid between B. napus and B. campestris, which has been explored practically in rapeseed production for many years. The experiment was first carried out on 65 trigenomic hybrids in 1999. The average over-mid-parent heterosis of biomass production was around 30%, and the highest value was 175.4%. In the following year, the observation was expanded to 120 trigenomic hybrids and the best average over-mid parent heterosis was 93%. A total of 1,477 DNA fragments, generated by Southern hybridization with 50 Brassica cDNA clones and 25 Arabidopsis EST clones, was scored across their parental lines. One hundred and twenty six and 215 fragments were identified as significantly associated with biomass production respectively in the 2 successive years. Using these active markers, a statistical model to resolve the heterosis is proposed and a new way to make use of the subgenomic heterosis is also discussed. PMID- 12582934 TI - One large-insert plant-transformation-competent BIBAC library and three BAC libraries of Japonica rice for genome research in rice and other grasses. AB - We report one large-insert BIBAC library and three BAC libraries for japonica rice cv Nipponbare. The BIBAC library was constructed in the HindIII site of a plant-transformation-competent binary vector (pCLD04541) and the three BAC libraries were constructed in the BamHI, HindIII and EcoRI sites of a BAC vector (pECBAC1), respectively. Each library contains 23,040 clones, has an average insert size of 130 kb, 170 kb, 150 kb and 156 kb, and covers 6.7x, 8.7x, 7.7x and 8.0 x rice haploid genomes, respectively. The combined libraries contain 92,160 clones in total, covering 31.1 x rice haploid genomes. To demonstrate their utility, we screened the libraries with 55 DNA markers mapped to chromosome 8 of the rice genetic maps and analyzed a number of clones by the restriction fingerprinting and contig assembly method. The results indicate that the libraries completely cover the rice genome and, thus, are well-suited for genome research in rice and other gramineous crops. The BIBAC library represents the first plant-transformation-competent large-insert DNA library for rice, which will streamline map-based cloning, functional analysis of the rice genome sequence and molecular breeding in rice and other grass species. These libraries are being used in the development of a whole-genome, BAC/BIBAC-based, integrated physical, genetic and sequence map of rice and in the research of genome-wide comparative genomics of grass species. PMID- 12582935 TI - Development of RAPD and SCAR markers linked to the Pvr4 locus for resistance to PVY in pepper ( Capsicum annuum L.). AB - Potato Virus Y (PVY) is the only potyvirus infecting pepper ( Capsicum annuum L.) in Europe. Currently, the development of pepper varieties resistant to PVY seems to be the most-efficient method to control PVY damage. Among the sources of resistance, a monogenic dominant gene Pvr4 confers resistance against all known PVY pathotypes. In this work, bulked segregant analysis (BSA) was used to search for randomly amplified polymorphic DNA (RAPD) markers linked to the Pvr4 gene, using segregating progenies obtained by crossing a homozygous resistant ('Serrano Criollo de Morelos-334') with a homozygous susceptible ('Yolo Wonder') cultivar. Eight hundred decamer primers were screened to identify one RAPD marker (UBC19(1432)) linked in repulsion phase to Pvr4. This marker was converted into a dominant sequence characterised amplified region (SCAR) marker (SCUBC19(1423)). This marker was mapped into a dense Capsicum genetic map in a region where several genes for resistance to different diseases are located. This marker can be useful to identify PVY-resistant genotypes in segregating progenies of pepper in marker-assisted selection (MAS) breeding programs. PMID- 12582936 TI - Genetic dissection of seed shattering, agronomic, and color traits in American wildrice ( Zizania palustris var. interior L.) with a comparative map. AB - A comparative map of American wildrice ( Zizania palustris var. interior L.) was used to identify loci controlling seed shattering, plant height, maturity, tiller number, plant habit, panicle length seed length, and color traits. Two to six significant quantitative-trait-loci (QTLs, P < 0.05) were detected for each trait evaluated, representing the first trait-mapping in wildrice. The chosen population was designed to emphasize the mapping of loci controlling the shattering trait, which is the most important trait in the management of this newly domesticated species. Three loci were detected that controlled the discretely categorized variation between shattering and non-shattering plants. Seed-shattering loci were detected and validated among the F(2) and F(3) generations. A multiple regression model with these three loci described 49.6% of the additive genetic variation. A genetic model with the same three loci including dominance and locus interactions predicted the shattering versus non shattering phenotype at a success rate of 87%. The comparative map was based on mapped RFLP markers used in white rice ( Oryza sativa L.) and other grass species. Anchor loci provided a reference point for the identification of potential orthologous genes on the basis of white rice mutant loci and consensus grass species QTLs. Candidate orthologous loci were identified among all traits evaluated. The study underscores the benefits of extending trait analysis through comparative mapping, as well as challenges of QTL analysis in a newly domesticated species. PMID- 12582937 TI - A genetic linkage map of willow ( Salix viminalis) based on AFLP and microsatellite markers. AB - The genus Salix (willow) contains a number of species of great value as biomass crops. Efforts to breed varieties with improved biomass yields and resistances to pests and diseases are limited by the lack of knowledge on the genetic basis of the traits. We have used AFLP and microsatellite markers to construct a genetic map of willow from a full-sib cross of the diploid species Salix viminalis (2n = 38). In accordance with a double pseudo-testcross approach, separate parental maps were constructed and merged to produce a consensus map comprising 291 AFLP and 39 willow microsatellite markers. Nineteen poplar microsatellites were also tested in willow. Five of these amplified loci, of which two were mapped. Linkage groups of the consensus map that could be identified in the parental maps are presented here and spanned 1,256.5 cM with an average interval between markers of 4.4 cM. PMID- 12582938 TI - Hypomethylation of CNG targets induced with dihydroxypropyladenine is rapidly reversed in the course of mitotic cell division in tobacco. PMID- 12582939 TI - [Frame-less and frame-based stereotaxy? How to choose the appropriate procedure]. AB - In recent years, additional tools for image guided surgery have been developed. Devices are now available for frameless punction and biopsy of intracerebral lesions and the indications for frameless and frame-based stereotaxy are increasingly overlapping. The objective of our paper was to analyse the most important factors that influence the decision on when to use frameless and when frame-based stereotaxy. Criteria such as application accuracy, image information and ergonomics were investigated for different lesions. Frameless and frame-based stereotaxy are complementary systems with different indications. They are different in terms of image information and ergonomics. Image guided surgery is the standard for continuous 3-dimensional topographical orientation. With high quality images (1 mm slice thickness) and bone marker registration, frameless stereotaxy may achieve an application accuracy comparable to frame-based systems. However, when using adhaesively mounted skin fiducials for patient registration, frameless stereotaxy is less accurate than framebased systems. Therefore, the size of the lesion should exceed 10 mm for punction and biopsy with a frameless system. Frame-based stereotaxy remains the gold standard for accurate targeting of lesions < 10 mm and for functional procedures. PMID- 12582940 TI - The use of postoperative ACTH levels as a marker for successful transsphenoidal microsurgery in Cushing's disease. AB - OBJECTIVES: The declines of ACTH and other POMC metabolites immediately after tumor extirpation do not predict the complete tumor removal of an ACTH-secreting pituitary adenoma in Cushing's disease. However, the pituitary surgeon should be in a position to evaluate the surgical result as soon as possible for the eventual planning of early repeat surgery. So far, subnormal serum cortisol levels after surgery are widely accepted as the criterion for cure. We investigated whether the early postoperative ACTH concentration is a reliable marker for the initial surgical outcome as well as the long-term remission. METHODS: In a prospective study, 147 patients undergoing primary transsphenoidal microsurgery for Cushing's disease between 1990 and 1996 were investigated. The early postoperative ACTH courses were reviewed and compared with the long-term outcome. ACTH measurements were performed immediately after tumor excision, 1, 2, 4, 8, and 12 hours later, and the following morning. Further ACTH levels were determined on various days of the hospital stay. Glucocorticoids were not given until hypocortisolism was proven. RESULTS: Ninety-five patients (65 %) presented with subnormal ACTH levels (< 10 ng/l) during their postoperative stay, of whom two patients (2 %) experienced recurrence of disease after 66 and 100 months. Of 29 procedures with early postoperative ACTH levels ranging from 10-20 ng/l (20 %), one patient received further treatment for persistent (3.5 %) and one patient for recurrent Cushing's disease (3.5 %). Of 12 patients (8 %) with early postoperative ACTH levels in between 20 to 30 ng/l, one patient received further treatment for persistent (8 %) and four patients for recurrent disease (33 %). ACTH levels of more than 30 ng/l, found in 11 patients (7 %), were accompanied by persistent (8 patients, 73 %) or recurrent (2 patients, 18 %) Cushing's disease. CONCLUSION: Subnormal (< 10 ng/l) or low normal (< 20 ng/l) postoperative ACTH levels within the first 7 days after surgery can be regarded as early markers for complete removal of an ACTH adenoma as well as indicators for long-term outcome in Cushing's disease. The risks of persistence or recurrence of Cushing's disease rise with the level of early postoperative ACTH values. Normal ACTH levels should result into further evaluation of the ACTH-cortisol secretion, for example by stimulation- or suppression tests. PMID- 12582941 TI - [Dural arteriovenous fistulas with intracranial hemorrhage: diagnostic and therapeutic aspects]. AB - Seven patients presented with intracranial hemorrhage due to arteriovenous dural fistula. Six patients showed intracerebral hemorrhage combined with subdural hematoma and intraventricular hemorrhage in one case respectively, and one patient had infratentorial subarachnoid hemorrhage. Location of the fistulae was frontobasal (n=2), tentorium (n=2), transverse sinus (n=2), and superior sagittal sinus (n=1). Angiography revealed reflux into cortical veins in all cases. Therapy was surgery in both cases with fistula of the anterior cranial fossa with good results. An endovascular intraarterial therapy was performed in a case with circumscribed fistula of the superior sagittal sinus, this patient developed a second dural fistula during follow-up. Two patients with tentorial fistulae had primary endovascular treatment complicated by infarction of both thalami in one case and a recurrence of the fistula in the other. In the last case the fistula was closed by surgery. Out of two patients with widespread fistulae of the transverse sinus one made a good clinical recovery and the other remained unchanged. In the first case definite closure of a remnant of the fistula was refused, in the second no further therapy was recommended. PMID- 12582942 TI - [Shunt operation versus endoscopic ventriculostomy in normal pressure hydrocephalus: diagnostics and outcome]. AB - In contrast to shunt operation the indication for an endoscopic ventriculostomy in patients diagnosed for normal pressure hydrocephalus is not scientifically established. From September 1997 to October 2001 we operated on 79 patients diagnosed for normal pressure hydrocephalus. Diagnosis was established by means of the intrathecal lumbal or ventricular infusion test, the cerebrospinal fluid tap test and MRI-CSF flow studies pre- and post-operatively. In 60 patients (76 %) we implanted a ventriculo-peritoneal shunt (Miethke Dual-Switch valve), and in 15 patients (19 %) we performed the endoscopic assisted third ventriculostomy. With our created NPH recovery rate and use of the clinical grading for normal pressure hydrocephalus created by Kiefer we compared the operative results of both patient groups. Immediately after the operation the results are the same for both treatments. In the follow-up examination after 12 and 27 months patients who underwent a ventriculostomy showed a better outcome, but the underdrainage rate was higher. Concerning the operation related complications the shunt treatment leaded to 10 revisions (17 %) because of four infections (7 %), two shunt insufficiencies (3 %), two overdrainages (3 %), two catheter dislocations (3 %). The ventriculostomy leaded to one case with a pneumatocephalus (7 %) and one ischemic thalamic lesion (7 %). In both operation methods we saw cases of underdrainages, three after valve implantation (5 %) and two after ventriculostomy (13 %). In that patients we performed a change of the implanted valve with a lower pressure level or rather an implantation of a valve system in the two cases who underwent a ventriculostomy. In patients with a pathologically increased resistance to CSF outflow in the lumbal infusion test a shunt implantation with the Miethke Dual-Switch valve is indicated. Patients whose outflow resistance is increased in the ventricular infusion test are suspected for a functional interventricular stenosis and should be treated by means of an endoscopic assisted ventriculostomy. PMID- 12582943 TI - [Prophylactic antibiotics in lumbar disc surgery: analysis of 1,030 procedures]. AB - OBJECTIVE: The purpose of this study was to investigate the efficacy of perioperative antibiotic administration in the prophylaxis of wound infection in lumbar disc surgery. METHODS: In 1989, 541 conventional lumbar discectomies were performed to treat nucleus pulposus prolapse in 533 patients at the neurosurgical department of the Benjamin-Franklin-Hospital (Free University of Berlin). Each patient received 2 g of the antibiotic Cefotiam intravenously at induction of anesthesia. During the previous year no antibiotic was administered in 636 similar operations (in 628 Patients). Acquisition of data was performed retrospectively. After statistical stratification there remained 492 procedures in 461 patients in the prophylaxis group and 538 procedures in 475 patients in the control group. Regarding patients age, duration of the surgical procedure and distribution of individual surgeons there were no significant differences between these two groups. RESULTS: The rate of infection was 0.2 % (n=1) in operations performed after antibiotic administration versus 2.8 % (n=15) in procedures without antibiotic prophylaxis. This difference is statistically significant (p < 0.0001). CONCLUSION: In accordance with the reviewed literature, this study confirms that one preoperative intravenous ('single shot') administration of Cefotiam is effective in decreasing the rate of postoperative wound infections in lumbar disc surgery. PMID- 12582944 TI - Age-related expression of p53, Mdm2, EGFR and Msh2 in glioblastoma multiforme. AB - OBJECTIVE: Glioblastoma multiforme (GBM) is a major cause of morbidity and mortality in neurosurgical patients. Despite the overall poor prognosis a range in survival times exists. Many approaches have been undertaken to define patient subgroups based on molecular changes. The aim of this study was to assess a possible correlation between the immunohistochemical p53, Mdm2, EGFR and Msh2 expression and age. METHOD: 143 patients (77 male, 66 female) were included in this retrospective study who underwent craniotomy for newly-diagnosed GBM between May 1994 and February 2000. For statistical analysis, patients were separated into three age groups: 1. < 40 years, 2. 40-60 years, 3. > 60 years. Immunohistochemical staining (IHC) was performed using anti-p53 (clone DO-1), anti-Mdm2 (clone IF-2), anti-EGFR (clone H11) and anti-Msh2 antibodies (clone AB 1). The results were compared with the Ki67/MIB-1 proliferation index (Ki67 PI) and patient survival. FINDINGS: P53 protein expression was significantly decreasing with advanced age (p < 0.05) whereas EGFR and Mdm2 expression was increasing (p < 0.05; p=0.01). Msh2 expression was unrelated to age. Multivariate analysis revealed Msh2 protein expression as a significant predictor of prolonged survival (p=0.004) whereas p53, Mdm2 and EGFR were not associated with patient survival. P53, Mdm2, EGFR and Msh2 expression was not associated with the Ki67 PI. INTERPRETATION: Our results support the hypothesis that in GBM patients a complex relationship exists between the p53, Mdm2 and EGFR expression and age. Msh2 expression is not related to age. Notably, nuclear Msh2 expression turned out to be an independent prognostic indicator. PMID- 12582945 TI - Physiological, subjective and performance effects of pseudoephedrine and phenylpropanolamine during endurance running exercise. AB - The aim of the study was to assess the effect of maximal therapeutic dosing of sympathomimetic amines found in over-the-counter (OTC) decongestant preparations on endurance running. Following familiarisation and a graded exercise test to determine maximal oxygen uptake (VO2 max), trained male runners (n = 8) completed four exercise sessions each separated by a minimum of one week. Each session was comprised of 20 min of sub-maximal treadmill running (70 % VO2 max) followed by a 5,000-m time trial on the treadmill under drug, placebo or control conditions. Drugs were administered in their commercial format over the 36-hour period prior to testing in the manufacturer's recommended maximal doses (i. e. 25 mg of phenylpropanolamine and 60 mg of pseudoephedrine four times daily). During sub maximal endurance running no statistical differences were observed in heart rate, VO2, minute ventilation, respiratory exchange ratio, blood lactate, glucose or non-esterified fatty acids (NEFA) or ratings of perceived exertion with respect to the treatment administered. Similarly there were no statistical differences according to the condition during the 5,000-m running time trial, in terms of heart rate, ratings of perceived exertion, time of completion and pre and post exercise blood lactate, glucose or NEFA. The results indicate that in maximal, multiple therapeutic doses both pseudoephedrine or phenylpropanolamine as present in common OTC decongestant formulations do not affect, nor possess any ergogenic properties with regard to, endurance running. PMID- 12582946 TI - Influence of acute moderate hypoxia on time to exhaustion at vVO2max in unacclimatized runners. AB - Eight unacclimatized long-distance runners performed, on a level treadmill, an incremental test to determine the maximal oxygen uptake (VO2max) and the minimal velocity eliciting VO2max (vVO2max) in normoxia (N) and acute moderate hypoxia (H) corresponding to an altitude of 2,400 m (PIO 2 of 109 mmHg). Afterwards, on separate days, they performed two all-out constant velocity runs at vO2 max in a random order (one in N and the other in H). The decrease in VO2max between N and H showed a great degree of variability amongst subjects as VO2max decreased by 8.9 +/- 4 ml x min(-1) x kg)(-1) in H vs. N conditions (-15.3 +/- 6.3 % with a range from -7.9 % to -23.8 %). This decrease in VO2max was proportional to the value of VO2max (VO2max vs. delta VO2max N-H, r = 0.75, p = 0.03). The time run at vVO2max was not affected by hypoxia (483 +/- 122 vs. 506 +/- 148 s, in N and H, respectively, p = 0.37). However, the greater the decrease in vVO2max during hypoxia, the greater the runners increased their time to exhaustion at vVO2max (vVO2max N-H vs. tlim @vVO2max N-H, r = -0.75, p = 0.03). In conclusion, this study showed that there was a positive association between the extent of decrease in vVO2max, and the increase in run time at vVO2max in hypoxia. PMID- 12582947 TI - Interleukin-6 expression after repeated bouts of eccentric exercise. AB - Plasma interleukin-6 (IL-6) is known to increase in response to eccentric exercise due to an acute-phase immune response. However, the severity of muscle injury is reduced with repeated bouts of eccentric exercise, possibly as a result of decreases in plasma IL-6. This study determined the response of IL-6 mRNA and IL-6, troponin-I (sTnI), muscle strength, and soreness as a result of repeated bouts of eccentric exercise. Eight males underwent two eccentric exercise bouts (3 wk apart) involving 7 sets of 10 repetitions at 150 % of the isotonic 1-RM of the dominant knee extensors. Blood samples were taken before, after and 2, 4, 6, 24, 48 and 96 h post-exercise. Strength and soreness ratings were assessed before and at 24, 48 and 96 h-post. Data were analyzed with 2 x 4 and 2 x 8 ANOVAs and the non-parametric Friedman test (p < 0.05). Both IL-6 mRNA and IL-6 underwent peak increases (p < 0.05) at 4 h-post and 6 h-post, respectively, but were not different between bouts. However, there were significant changes (p < 0.05) in sTnI, strength, and soreness that were greater after the first bout than the second, characteristic of the repeated bout effect. These results indicate that changes in sTnI, strength and soreness were less with the second eccentric exercise bout whereas the changes in both IL-6 mRNA and protein were not effected between bouts. PMID- 12582949 TI - Influence of chronic aerobic exercise on microcirculatory flow and nitric oxide in humans. AB - In the present study we assessed the effect of physical training on Laser Doppler skin flux (LDF) and nitric oxide (NO) release, before and after 3 min of brachial artery occlusion. To this end we performed laser Doppler measurements and the venous plasma assay of nitrite/nitrate (NOx) on 10 sedentary healthy subjects and 10 endurance athletes. The sedentary control subjects had lower basal and post reperfusion levels of NOx as compared to athletes (mean +/- SE: 27.8 +/- 3.5 vs. 33.2 +/- 3.4, 48.6 +/- 7.9 vs. 60.1 +/- 10.1 micromol/L; p < 0.05). LDF at baseline was not significantly different in the two groups (157.5 +/- 7.9 and 176.64 +/- 26.7 PU for sedentary subjects and athletes, respectively) while post ischemic LDF was significantly lower in nonathletic subjects than in athletes (209.9 +/- 13 and 343.8 +/- 21.3 PU, p < 0.001). In both groups the hyperaemic stimulus significantly increased LDF and NOx levels (p < 0.01 and p < 0.05, respectively). The flow reserve, estimated as peak/basal LDF, was significantly lower in control subjects than in athletes (1.34 +/- 0.2 and 2.32 +/- 0.9, respectively, p < 0.01). In athletes, as opposed to sedentary subjects, a direct correlation was found between plasma NOx concentration and LDF both in basal conditions (r = 0.92; p < 0.001), and during hyperaemia (r = 0.84; p < 0.01). In conclusion, compared to sedentary subjects, athletes had an enhanced nitric oxide release. Hyperaemia increased LDF and nitric oxide levels both in sedentary subjects and in athletes. PMID- 12582948 TI - Physical activity, hormone replacement therapy and plasma lipoprotein-lipid levels in postmenopausal women. AB - We assessed the cross-sectional associations between hormone replacement therapy (HRT), habitual physical activity levels and plasma lipoprotein-lipid levels in postmenopausal women. Sedentary (n = 19), active nonathlete (n = 20) and endurance-trained (n = 21) postmenopausal women, with half of each group on and half not on HRT, underwent assessments of plasma lipids, VO2max, body composition, diet and common genetic variants. The groups' physical characteristics were generally similar though body weight was higher in the active nonathletes, and body fat was lower and VO2max higher in the athletes. HRT was associated with beneficial total cholesterol, LDL-C and HDL-C levels in sedentary and active nonathlete women. Plasma lipoprotein-lipid profiles were similar in women athletes on and not on HRT, with their profiles being only slightly better than sedentary and physically-active women on HRT. After controlling for HRT status, VO2max was correlated with total cholesterol (r = 0.51, p = 0.0001), LDL-C (r = -0.52, p = 0.0001), HDL-C (r = 0.25, p = 0.055), HDL 2 -C (r = 0.24, p = 0.08) and TG levels (r = -0.46, p = 0.0001). After controlling for HRT status, % body fat was correlated with total cholesterol (r = 0.43, p = 0.001), LDL-C (r = 0.38, p = 0.003), HDL-C (r = -0.29, p = 0.025), HDL 2 -C (r = -0,26, p = 0.07) and TG levels (r = 0.40, p = 0.002). Dietary fat intake was similar among the groups. APO E genotype was only associated with plasma lipid profiles in athletes, as those with at least one APO E2 allele tended to have better lipid profiles than those with only APO E3 or E4 alleles. Thus, HRT, exercise training and body composition are associated with plasma lipid levels in postmenopausal women; common polymorphic variations at key lipid metabolism-related gene loci also may interact with exercise training to affect their plasma lipid profiles. PMID- 12582950 TI - The biological and metabolic adaptations to 12 months training in elite rowers. AB - We studied the effects of a twelve months endurance-training program on exercise induced change in blood contents in thirteen rowers. A standardized testing session (18 km rowing at 80 % of VO2max) was performed 19 times during the training program. Capillary blood samples were taken at rest and immediately post exercise to analyse a wide range of serum concentrations. During exercise, glucose and lactate concentrations stabilized after only five training weeks and did not evolve from that point. Transport and hepatic protein concentrations increased with exercise up to the 15th week (p = 0.03), and remained stable from that point (p = 0.02). Evolution of exercise-induced change in alpha 1 -acid glycoprotein concentration revealed protein metabolism adaptations to training. Change in alpha 1 -acid glycoprotein concentrations were exactly opposite to that of urea and alpha 1 -antitrypsin (p = 0.01 and 0.002, respectively). Immunoglobulin concentrations exhibited important increases up to the 6th training week (p < 0.05), and a global stabilization was observed from that point. However, analysis of IgG subclasses highlighted significant changes that could not be found with the study of total IgG concentrations. Evolution of the exercise-induced change in Apo-A 1 /Apo-B concentrations ratio was also more informative about lipid metabolism than the Apo-A 1 and Apo-B concentrations taken individually. Indeed, evolution of metabolic changes during exercise should be carefully monitored during training to avoid interpretative errors on the training status of athlete. PMID- 12582951 TI - Ability of different physical activity monitors to detect movement during treadmill walking. AB - This study assessed the ability of four different activity monitors to discriminate changes in treadmill walking velocity. The relationships between walking velocity and bodily movement and between bodily movement and energy expenditure determined by indirect calorimetry (IC-EE or METs) were determined. Twenty-eight subjects walked at 3.2, 4.0, 4.8, 5.6, and 6.4 km/h (0 % grade) for 30 min on separate occasions. The Tritrac-R3D (TT), Computer Science & Applications, Inc. (CSA), and Mini-Logger (ML) activity monitors that measure bodily acceleration in one or three planes, and a Yamax Digiwalker-500 (YX) that records footsteps, were secured at the waistline of each subject. CSA monitors were also worn at the wrist and ankle. Walking velocity and bodily movement were significantly related (r = 0.89 to 0.93) for TT, CSA, ML, and YX. Importantly, changing each walking velocity produced significant changes in bodily movement that was detected by each monitor. Bodily movement and IC-EE were significantly related for TT, CSA, ML, and YX (r = 0.47 to 0.94). Compared to IC-EE, and at all walking speeds, EE was significantly overestimated by the TT, and EE was significantly underestimated by the YX. These results indicate that the activity monitors can differentiate bodily movement associated with walking at slow speeds better than they can estimate energy expenditure associated with walking at slow speeds. PMID- 12582952 TI - Maximal voluntary eccentric, isometric and concentric torque recovery following a concentric isokinetic exercise. AB - To examine neuromuscular fatigue and recovery following an isokinetic fatiguing exercise, nine active females performed a fatiguing exercise comprising of ten sets of ten maximal concentric knee extensions. Before (pre-test), five minutes (post-test), 24 h and 48 h after the fatiguing exercise, maximal voluntary eccentric (-1.05 rad x s(-1); -2.09 rad x s(-1)), isometric (0 rad x s(-1)) and concentric (1.05 rad x s(-1); 2.09 rad x s(-1)) torque were measured. In order to distinguish central from peripheral factors involved in torque decrement, activation level (twitch interpolation technique) and twitch contractile properties were recorded. During the course of the fatiguing exercise, concentric torque was significantly lower during the 3rd set than pre-test (-5.6 +/- 12.3 %) and further decreased to the 10th (-10.3 +/- 9.5 %). Eccentric and isometric torques were significantly lower during post-test than pre-test (-16.8 +/- 8.8 % at -2.09 rad x s(-1), -15.1 +/- 7.4 % at -1.05 rad x s(-1), and -10.4 +/- 5.9 % at 0 rad x s(-1); p < 0.05), while concentric torque was not significantly modified. Voluntary activation, peak twitch torque, twitch maximal rates of force development and relaxation were also significantly declined (p < 0.05) at post test. Twenty-four hours later, all the measured parameters were close to their pre-fatigue values. The present results reveal that the best way to test concentric-induced alteration of neuromuscular function was to use stressful testing conditions, such as eccentric contractions. PMID- 12582953 TI - Familial aggregation of exercise heart rate and blood pressure in response to 20 weeks of endurance training: the HERITAGE family study. AB - Changes of heart rate (HR) and blood pressure (BP) relative to baseline levels in response to an extended period of endurance training are indices of cardiovascular adaptability. Familial influences were investigated for HR and BP at work rates of 50 W and 60 % of the maximal oxygen uptake (VO2max) in response to 20 weeks of endurance training. A total of 481 participants from 99 sedentary White nuclear families in the HERITAGE Family Study (HERITAGE) were analyzed using a familial correlation model. Each of these training response phenotypes was adjusted for the effects of age, BMI, cigarette smoking, baseline VO2max, and its baseline values in fathers, mothers, sons and daughters, respectively. We found that maximal heritabilities reached 34 % and 29 % for HR training responses at 50 W and 60 % of VO2 max, respectively. The heritability was 22 % for systolic BP (SBP) training response at 50 W, but negligible at 60 % of VO2max. No significant heritabilities were found for diastolic BP (DBP) training responses at either 50 W or 60 % of VO2max. Familial influences for exercise HR and BP training responses were also assessed in a total of 257 participants from 113 Black family units in HERITAGE. However, there was no significant familial resemblance, which may be attributable to the small sample size. In conclusion, HR and SBP training responses during submaximal exercise in Whites were influenced by a modest, but significant, familial component. These observations are therefore in contrast to substantial familial effects (heritability estimates of about 50 %) previously reported for these variables measured at baseline. PMID- 12582954 TI - The effects of prior cycling and a successive run on respiratory muscle performance in triathletes. AB - The aim of the present study was to compare the effects of prior cycling and a successive run on respiratory muscle performance during a cycle-run succession as performed in the triathlon. We hypothesized that despite the moderate intensity of exercise and the absence of exhaustion, the crouched cycling position would induce a decrease in respiratory muscle performance that would be reversed by the successive vertical run position. Ten male triathletes (22.6 +/- 1.1 yr) performed a four-trial protocol: (1) an incremental cycle test to assess maximal oxygen uptake (VO2max), (2) 20 min of cycling (C), (3) 20 min of running (R), and (4) 20 min of cycling followed by 20 min of running (C-R). Trials 2, 3 and 4 were performed at the same metabolic intensity, i. e., 75 % of VO2max. Respiratory muscle force was assessed by measuring maximal expiratory (P(Emax)) and inspiratory (P(Imax)) pressures from the functional residual capacity (FRC) before and 10 min after C, R, and C-R. Respiratory muscle endurance was assessed one day before and 30 min after C, R, and C-R, by measuring the time limit (T(lim)), which corresponds to the length of time a respiratory load can be sustained before the process of fatigue develops sufficiently to cause task failure. The results showed a similar significant decrease in P(Imax) (132.4 +/- 4.9 versus 125.7 +/- 5.6 cm H2O, p < 0.05) and T(lim) (5.22 +/- 0.28 versus 3.68 +/- 0.32 min, p < 0.05) post-C and post-C-R (133.7 +/- 4.0 versus 126.9 +/- 5.2 cm H2O, and 5.29 +/- 0.18 versus 3.49 +/- 0.41 min, respectively, p < 0.05) compared with the pre-trial values. In contrast, P(Imax) and T(lim) were not significantly decreased post-R (131.8 +/- 6.1 cm H2O versus 129.6 +/- 6.4 cm H2O, and 4.90 +/- 0.69 versus 4.40 +/- 0.56 min, respectively, p > 0.05). We concluded that moderate intensity exercise not performed to exhaustion induced a decrease in respiratory muscle performance. Moreover, the respiratory muscle fatigue induced by prior cycling was maintained, and neither reversed nor worsened, by the successive run. PMID- 12582955 TI - Improved muscle strength and power in elderly exercising regularly. AB - Since the number of elderly people with a sedentary lifestyle is considerable and still growing, regular exercise could be an extra stimulant to compensate for the decrease in functional capacities. The potential positive effects of physical and sporting activities (PSA) on muscular function in relation to the period of practice were studied. Muscular strength and power were evaluated by dynamometric knee and ankle tests on 40 healthy subjects aged over 60 years old, split into four groups according to the period of practice. Higher muscular performance was obtained in subjects who had always practised PSA, whereas subjects who had always been inactive displayed the worst muscular performances. Subjects who had begun PSA practice lately had good muscular strength and power, close to those of the AA group, whereas the subjects who had stopped the practice at an early age did not perform as well. Life-long PSA attenuates the age-related loss of muscle function, and initiation at old age improves it. This, and the absence of any beneficial effects of PSA stopped when young, indicates that the actual activity level is an important determinant of skeletal muscle function at old age. Practicing PSA increases muscular strength and power and thus improves daily living activities in elderly people. PMID- 12582956 TI - Breathing and propelling in crawl as a function of skill and swim velocity. AB - This study analyzes apnea (A), exhalation (E), and inhalation (I) duration with respect to stroke organization in front crawl as a function of inhalation side, swim velocity and performance level. Thirty-six male subjects comprised two groups based on performance level: more expert (ME) and less expert (LE) swimmers. All swam with one inhalation per cycle to the preferred side at speeds corresponding to two specific race paces: 100-m (V100) and 800-m (V800) velocities. The breathing arm (BA) is located on the inhalation side, and the non breathing arm (NBA) on the opposite side. The sound of air passing in and out of the swimmers' mouths was captured by a microphone and synchronized with video frames. Stroke phases and arm coordination were identified by video analyses. Arm coordination was quantified using two indices of coordination (IdC) corresponding to the lag time between the beginning of the BA (IdC-BA) or NBA (IdC-NBA) propulsive action and the end of that of the other arm. As velocity increases, the ME are observed to reduce I during BA recovery (-19.4 +/- 31.6 %, p < 0.05) while the LE increase A (+ 34.8 +/- 25.2 %, p < 0.05) during BA entry, catch and recovery and NBA pull and push. These variations are related to a lengthening of the pull for both arms at the expense of BA non-propulsive phases. At V100, the ME have greater E (p < 0.05) during BA entry and catch (+ 21.1 +/- 38.2 %) and NBA push (+ 26.3 +/- 39.5 %) compared to the LE. This increase, at the expense of A, corresponds to a shorter BA push and NBA recovery. At V800, the ME exhibit a longer A (p < 0.05) during BA recovery (+ 19.9 +/- 33.2 %) and NBA pull (+ 24.2 +/- 31.5 %), and decreased I during NBA push and pull. These differences are related to a shortening of BA recovery and pull and a longer push for both arms. These breath and stroke adaptations correspond to an increase in stroke rate and IdC-BA with the velocity and performance level. This study points out the breathing-propelling aspects of coordination that indicate technical skill in swimming. PMID- 12582958 TI - Oblique radial forearm reverse-flow flap. AB - A modifed design for the distally-based radial forearm flap is presented, in an oblique direction rather than longitudinally, based on the existence of skin laxity in the proximal forearm region. The skin paddle of the flap is designed in an oblique fashion pedicled on one of the proximal-row septocutaneous perforators, and elevated in the usual manner supplied by the distal radial artery. The oblique radial forearm flap thus created was successfully utilized for reconstruction of seven dorsal hand defects. Results showed that all the flaps could easily be transposed to the defect through a wide arc of rotation and all survived totally, with direct closure of the donor site in five cases, and significant reduction in size in the remaining two cases. It was concluded that the oblique design for the skin island of the reverse radial forearm flap could allow creation of a flap that has a smaller donor defect and yet presents a longer pedicle length, with a wider arc of rotation and better adaptation to a dorsal hand defect, than a conventional longitudinal-design radial forearm flap. PMID- 12582959 TI - Periarterial sympathectomy salvage of the acutely ischemic hand. AB - A 40-year-old woman involved in a motor vehicle accident presented to the emergency room with signs and symptoms of acute left hand ischemia. The mechanism of injury consisted of a severe crushing component limited to a linear zone across the left hand metacarpals. After patient management, and compartment decompressions and stabilization of fractures, an ischemic state persisted that failed to improve after extensive vessel exploration and bathing in vasodilating solutions. Only after extensive peripheral sympathectomy was appropriate flow re established to the hand. The need to employ periarterial sympathectomy in the acute trauma setting will occur infrequently, but is a valuable tool to have in mind when flow is not re-established after appropriate less invasive measures have failed. PMID- 12582961 TI - Telescoping vascularized fibular graft: a new method. AB - The authors describe a case of congenital pseudarthrosis of the tibia treated with a telescoping vascularized fibular graft. The advantages of this new method are that bone union and leg-length discrepancy may be corrected simultaneously. PMID- 12582962 TI - Nerve anastomosis with glue: comparative histologic study of fibrin and cyanoacrylate glue. AB - The authors report the results of a comparative experimental nerve study, using a biologic tissue glue (fibrin) and a synthetic glue (2-cyanoacrylate) in a rat model. A tension-free repair is necessary with the use of fibrin glue, or gapping may occur, thus limiting the use of the agent in promoting re-neurotization. In addition, the human origin of fibrin and thrombin allow for the possibility of viral transmission. The aim of the study was to verify if the synthetic glue is a viable alternative, or whether it causes cellular and tissue lesions. Their main finding was that the cyanoacrylate causes a foreign-body inflammatory reaction and retractile fibrosis, often reducing the nerve diameter up to two-thirds. Cyanoacrylate glue is thus not recommended for peripheral nerve repair. PMID- 12582963 TI - Limits of muscle-to-nerve ratio in functional muscle transplantation. AB - The aim of the present study was to investigate the functional limits of over dimensioning a free functioning muscle transplant for neuromuscular reconstruction. A part of the rectus femoris muscle, two and three times bigger than a scutuloauricularis muscle, was used to functionally replace the latter after removal. The major finding of these experiments is that both double-sized and triple-sized portions of the rectus femoris muscle developed maximal tetanic tensions during isometric contractions, which were up to 175 percent of the control scutuloauricularis muscle of the unoperated, contralateral side, although the same branch of the facial nerve was used for reinnervation of the grafted muscle. This implies that the supplying branch of the facial nerve has the potency to innervate a muscle much larger than the originally supplied muscle with optimal efficiency. These results underline the usefulness of overdimensioning during functional muscle transplantation, and also in limited neural capacity situations. PMID- 12582964 TI - Effects of hyaluronic acid-carboxymethylcellulose membrane on extraneural adhesion formation and peripheral nerve regeneration. AB - The goal of nerve repair in the peripheral nervous system is to increase the number of axons passing from proximal to distal stump, and to enable the regenerated axons to reach the end organ as soon as possible. In the present study, the effect of the membrane formed by a mixture of hyaluronic acid and carboxymethylcellulose (HA-CMC) on nerve regeneration and perineurial scar formation was investigated. Eighteen New Zealand rabbits were allocated into control (n = 9) and experimental groups (n = 9). In the control group, conventional nerve repair was carried out following the transection of the sciatic nerve, while in the experimental group, following repair of the nerve, the repair line was covered by HA-CMC membrane extending 1 cm beyond the distal and proximal ends. Nerve regeneration and extraneurial adhesion formation were compared between the two groups 3 months later. It was observed that adhesion in the surrounding tissues was significantly less in the experimental group than in the control group. Furthermore, morphometric analysis of specimens obtained from the distal parts of nerves showed that the number of axons with myelin was higher in the experimental group than in the control group, with a statistically significant difference. Histologic sections obtained from the nerve repair line demonstrated that extraneural and intraneural fibrosis was significantly lower in the experimental group. It was concluded that HA-CMC membrane had a favorable effect on nerve regeneration, as well as extraneural scar formation, encouraging the clinical application of HA-CMC following nerve injuries. PMID- 12582965 TI - Using human hair as suture material in microsurgical practice. AB - For practice in microsurgery, animal models and microsurgical instruments are required. The use of some synthetic and non-animal materials such as human placenta and other tissues have also been suggested for practicing. The other material that is needed is microsuture. These materials are expensive and often not easily obtained. To meet this requirement, the authors present human hair as a microsuture material for microsurgical practice. PMID- 12582966 TI - Functional recovery and histomorphometric assessment following tibial nerve injury in the mouse. AB - Longitudinal studies have established that functional recovery following sciatic nerve injury can be evaluated in the mouse. Injury to the tibial nerve offers several advantages to sciatic nerve injury, including improved lower extremity sensation and end-organ reinnervation. Functional recovery following tibial nerve crush injury was studied in 55 C3H mice randomized into five groups harvested for histomorphometric evaluation from either normal nerves or 2, 3, 4, or 6 weeks postoperatively. Walking tracks were obtained preoperatively, and at regular intervals postoperatively, and foot print lengths measured. Significant normalization of print length occurred 14 days postoperatively, and complete recovery was noted 28 days postoperatively. Significant histomorphologic evidence of neuroregeneration was detected between 2 and 4 weeks postoperatively. Injury to the tibial nerve is a viable alternative to the sciatic nerve for studying neural regeneration in mice, and the print length factor can be used to monitor functional recovery in this model. PMID- 12582967 TI - Orthotopic hindlimb transplantation in the mouse. AB - There are many potential advantages to using a mouse as a model for composite tissue transplantation, particularly to examine the mechanisms behind various tolerance induction protocols. The purpose of this study was to evaluate the reliability of a new hindlimb transplant model in the mouse. Fifteen Swiss Webster mice, 14 to 16 weeks old (35 to 50 g), had syngeneic hindlimb transplants performed. The donor limb was attached to the recipient limb stump first by bony fixation using an intramedullary rod (22-gauge needle, 2.5 mm in length). The venous anastomosis was performed across a stent fashioned from a 27-gauge nylon I.V. catheter tip cover and secured with two simple ties using 10-0 nylon. The arterial anastomosis was performed with interrupted 11-0 nylon stitches. Eleven of the fifteen transplanted limbs survived for the duration of the study (30 days). All four failures occurred within 4 hr postoperatively, (venous thrombosis, n = 2; arterial thrombosis, n = 2). Clamp time decreased throughout the study, averaging 56 min for the final four transplants and 88 min total operative time. Orthotopic hindlimb transplantation can be reliably achieved in a mouse. The mouse model should be useful to the future study of composite tissue allotransplantation. PMID- 12582969 TI - Role of nitric oxide in the mechanism of preclamping and remote ischemic preconditioning of adipocutaneous flaps in a rat model. AB - The purpose of this study was to determine whether nitric oxide (NO) plays a role in the mechanism of acute ischemic preconditioning (IP). Fifty-eight male Wistar rats were divided into seven experimental groups. An extended epigastric flap was raised in one of the control groups (C, n = 8), and a 3-hr flap ischemia was induced. Another group served as a non-ischemic control (CO, n = 8). The animals of group S (n = 9) received 500 nmol/kg of Spermine/Nitric Oxide Complex (Sper/NO) intravenously 30 min prior to ischemia. The group N+P (L-NAME + preclamping, n = 8) received 10 mg/kg Nomega-Nitro-L-Arginine Methyl Ester (L NAME) intravenously before preclamping of the flap pedicle (10-min cycle length, 30-min reperfusion). Ten mg/kg L-NAME were administered in group N+T (L-NAME + tourniquet, n = 9) before ischemia of the right hindlimb was induced using a tourniquet for 10 min after flap elevation. Flap ischemia was induced after 30 min of limb reperfusion. A similar protocol was used in the groups N+P+S (L-NAME + preclamping+Sper/NO, n = 8) and N+T+S (L-NAME + tourniquet + Sper/NO, n = 8). In both groups Sper/NO was administered 30 min prior to flap ischemia, additionally to the protocol of the groups N+P and N+T. Mean flap necrosis area was assessed on the fifth postoperative day using a planimetry software. Average flap necrosis area was 67 +/- 16 percent in the control group C, 28 +/- 13.3 percent in the non-ischemic controls (CO), 10 +/- 5.9 percent in group S, 77.5 +/ 10.2 percent in group N+P, 76 +/- 6.9 percent in group N+T, 71.5 +/- 9.4 percent in group N+P+S, and 78 +/- 9.9 percent in group N+T+S. The animals of group S and CO demonstrated a significantly lower area of flap necrosis than all other groups ( p < 0.001). No significant difference could be shown between the groups C, N+P, N+T, N+P+S and N+T+S. Group S showed a significantly lower flap necrosis area than group CO ( p < 0.01). The data showed, that NO plays an important role in the mechanism of IP since the administration of an NO-donor previous to ischemia simulates the effect of IP, while the unspecific blocking of NO synthesis by L NAME eliminates the protective effect of flap preconditioning by preclamping as well as by remote IP. Exogenous NO application is insufficient to provide protection once the endogenous NO synthesis is blocked. PMID- 12582970 TI - A psychopathological marker or biological psychiatry. PMID- 12582971 TI - The World Federation of Societies of Biological Psychiatry (WFSBP) Guidelines for the Biological Treatment of Bipolar Disorders, Part II: Treatment of Mania. AB - Identical to the preceding guidelines of this series, these practice guidelines for the biological, mainly pharmacological treatment of acute bipolar mania were developed by an international Task Force of the World Federation of Societies of Biological Psychiatry (WFSBP). Their purpose is to supply a systematic overview of all scientific evidence pertaining to the treatment of acute mania. The data used for these guidelines have been extracted from a MEDLINE and EMBASE search, from recent proceedings of key conferences, and from various national and international treatment guidelines. Their scientific rigor was categorised into four levels of evidence (A-D). As these guidelines are intended for clinical use, the scientific evidence was finally not only graded, but has also been commented by the experts of the task force to ensure practicability. Key words: bipolar disorder, mania, acute treatment, evidence-based guidelines, pharmacotherapy, antipsychotics, mood stabiliser, electroconvulsive therapy. PMID- 12582972 TI - Testosterone deficiency and mood in aging men: pathogenic and therapeutic interactions. AB - In contrast to women, men do not experience a sudden cessation of gonadal function comparable to menopause. However, there is a progressive reduction in hypothalamic-pituitary-gonadal (HPG) function in aging men: testosterone (T) levels decline through both central (pituitary) and peripheral (testicular) mechanisms and there is a loss of the circadian rhythm of T secretion. In cohorts of men 75 years of age, mean plasma T levels are 35% lower than comparable young men, and more than 25% of men over 75 appear to be T-deficient. Such age associated T deficiency, which has been termed 'andropause', is thought to be responsible for a variety of symptoms experienced by elderly men, such as weakness, fatigue, reduced muscle and bone mass, impaired haematopoiesis, oligospermia, sexual dysfunction, depression, anxiety, irritability, insomnia and memory impairment. However, it has been difficult to establish correlations between these symptoms and plasma T levels. Nevertheless, there is some evidence that T replacement leads to symptom relief, particularly with respect to muscle strength, bone mineral density, and haematopoiesis. Studies to date on the specific association between psychiatric symptoms, such as depressed mood, and T levels have been methodologically flawed. Overall, data suggest that although hypogonadism is not central to major depressive disorder (MDD), HPG hypofunction may have aetiological importance in mild depressive conditions, such as dysthymia. PMID- 12582973 TI - Pharmacokinetic Interactions between Antiepileptic and Antidepressant Drugs. AB - The use of antiepileptic drugs (AEDs) for the treatment of psychiatric disorders has reached a new phase of clinical interest. They are commonly used in the therapy of psychoses, mood disorders, aggression and eating disorders. Pharmacotherapy combinations involving AEDs and antidepressant drugs are used to treat co-morbid psychiatric and neurological disorders, to reduce or control the adverse effects of a medication or to increase a medication effect. Therefore, the impact of pharmacokinetic interactions of this class of drugs is quite relevant. In this paper, the available data about the mechanisms of metabolic kinetic interactions between antidepressant and antiepileptic drugs, as well as their clinical significance, has been reviewed. PMID- 12582974 TI - Serotonin transporter gene polymorphism and schizoid personality traits in the patients with psychosis and psychiatrically well subjects. AB - BACKGROUND AND OBJECTIVES: serotonin transporter (5-HTT) gene allelic variants were shown to be associated with Neuroticism and Harm Avoidance but the results were not replicated in other studies. The current investigation was undertaken in a further attempt to study the relationship between 5-HTT polymorphism and personality traits. SUBJECTS AND METHODS: to evaluate a spectrum of personality traits, MMPI was administered to a sample including patients with affective disorders (n=114), patients with schizophrenia spectrum illnesses (n=110) and psychiatrically well controls (n=124). All groups were genotyped for VNTR-17 and functional insertion-deletion (5-HTTLPR) polymorphisms. RESULTS: an association was found between 5-HTTLPR polymorphism and scores on three MMPI scales: Psychopathic deviance, Paranoia and Schizophrenia in patients with affective disorders and S chizophrenia in normal subjects. Both affected and control individuals with 'ss' genotype exhibited lower scores on these scales. CONCLUSION: we demonstrated that functional deletion/insertion allelic variation associated with decreased expression of serotonin transporter ('s' allele or 'ss' genotype) may restrict expression of schizoid traits in normal subjects and patients with affective disorders. PMID- 12582975 TI - A double-blind, placebo-controlled trial of clonazepam in obsessive-compulsive disorder. AB - Selective serotonin reuptake inhibitors (SSRIs) are currently the first-line pharmacological agents in treating obsessive-compulsive disorder (OCD). Appropriate treatment for OCD also involves cognitive behavioural therapy (CBT), including exposure and response prevention. As there is a time delay in seeing full therapeutic response, and not all patients tolerate SSRIs, there remains an unmet need for additional treatment approaches in OCD. In addition, most responders report only a partial reduction in symptoms. Clonazepam has demonstrated effectiveness in several preliminary reports in treating OCD. Twenty seven patients with OCD were entered into a 10 week, double-blind, parallel design trial of clonazepam vs. placebo. Overall, only 3 out of 25 patients who had >/= 1 rating on clonazepam/placebo were judged to be treatment responders, by scoring a 1 (very much improved) or 2 (much improved) on the CGI improvement scale. Responders included 2 of 9 in the placebo group and 1 of 16 in the clonazepam group. No significant difference was found between clonazepam and placebo groups on responder/non responder status (Chi(2 )=1.39, df =1,24, p=0.238), nor on change in YBOCS, Ham-A, Ham-D or NIMH scales from beginning to last evaluation carried forward. These findings suggest that clonazepam is not effective as monotherapy in treating OCD. Its effectiveness in specific subgroups of OCD patients with co-morbid anxiety disorders or as an augmentation strategy added to SSRIs remains to be determined. PMID- 12582976 TI - Self-rated aggression related to serum testosterone and platelet MAO activity in female patients with the fibromyalgia syndrome. AB - To investigate self-rated aggression in relation to platelet MAO activity and serum testosterone in patients with fibromyalgia syndrome (FMS), we administered the Aggression Questionnaire - Revised Swedish Version (AQ-RSV) to 30 female patients with FMS. After correction for age, significant positive correlations were seen between serum testosterone concentrations and the AQ-RSV scores for Verbal Aggression (r=0.36, p<0.05) and Anger (r=0.37, p<0.05), whereas the platelet MAO activity was negatively correlated with the score for Verbal Aggression (r=-0.44, p<0.05). Our results suggest that aggression and irritability in female FMS patients might be increased by elevated testosterone concentrations in combination with reduced capacity of the serotonergic system as reflected by low platelet MAO activity. PMID- 12582977 TI - Quetiapine in the treatment of borderline personality disorder. AB - Affective dysregulation, impulsivity and cognitive-perceptual difficulties are the psychopathological nuclear dimensions of Borderline Personality Disorder (BPD). Psychopharmacological treatment may become necessary during episodes of acute decompensation in which suicidal or self-destructive behaviour erupts. Some classes of psychotropic drugs have demonstrated efficacy in diminishing symptom severity and optimising functioning, such as antidepressants, mood stabilizers, benzodiazepines, opiate antagonists and antipsychotics. Conventional antipsychotics are the best-studied psychotropic medications for BPD, but nonadherence is often due to their severe side effects. Preliminary data reveal efficacy of atypical antipsychotics in BPD. We describe the impact of the novel antipsychotic drug quetiapine on severe self-mutilation in two female patients with the diagnoses of BPD. In both cases, monotherapeutic treatment with quetiapine was well tolerated and resulted in a marked improvement of impulsive behaviour and, over time, overall level of function. Though promising, our findings have to be regarded as preliminary. Due to the overall paucity of data there still is insufficient evidence to make a strong recommendation concerning continuation and maintenance therapy with atypical antipsychotics in BPD. Thus, there is a clear need for further controlled studies to evaluate pharmacological treatment options for this disorder. PMID- 12582978 TI - Automatic 3D registration for interventional MRI-guided treatment of prostate cancer. AB - The goal of this research is to register real-time interventional magnetic resonance imaging (iMRI) slice images with a previously obtained high-resolution MRI image volume, which in turn can be registered with functional images such as those from SPECT. The immediate application is in iMRI-guided treatment of prostate cancer, where additional images are desired to improve tumor targeting. In this article, simulation experiments are performed to demonstrate the feasibility of slice-to-volume registration for this application. We acquired 3D volume images from a 1.5-T MRI system and simulated low-field iMRI image slices by creating thick slices and adding noise. We created a slice-to-volume mutual information registration algorithm with special features to improve robustness. Features included a multiresolution approach, two similarity measures, and automatic restarting to avoid local minima. To assess the quality of registration, we calculated 3D displacements on a voxel-by-voxel basis over a volume of interest between slice-to-volume registration and volume-to-volume registration, which was previously shown to be quite accurate. More than 800 registration experiments were performed on MR images of three volunteers. The slice-to-volume registration algorithm was very robust and accurate for transverse slice images covering the prostate, with a registration error of only 0.4 +/- 0.2 mm. Error was greater at other slice orientations and positions. The automatic slice-to-volume mutual information registration algorithm is robust and probably sufficiently accurate to aid in iMRI-guided treatment of prostate cancer. PMID- 12582979 TI - Fast surface and volume rendering based on shear-warp factorization for a surgical simulator. AB - Fast simultaneous visualization of 3D medical images and medical instruments is necessary for a surgical simulator. Because unconstrained motion of a medical instrument is more frequent than that of the patient, the visualization of medical instruments is performed in real time using surface rendering. However, volume rendering is usually used for realistic visualization of the 3D medical image. We have developed an algorithm to combine a volume-rendered image and a surface-rendered image using a Z-buffer for depth cueing, which is applied to a surgical simulator. Surface rendering is used for visualization of a medical instrument, whereas 3D medical images such as CT and MRI are usually visualized by volume rendering, because segmentation of the medical image is difficult. In this study, when the volume-rendered image is combined with the surface-rendered image, the amount of computation is reduced by early ray termination and instrument-region masking in the sheared image space. Using these methods, a fast combination of volume-rendered and surface-rendered images is performed with high image quality. The method is appropriate for real-time visualization of 3D medical images and medical instrument motion in the images, and can be applied to image-guided therapy and surgical simulators. PMID- 12582980 TI - Validation of fluoroscopy-based navigation in the hip region: what you see is what you get? AB - Fluoroscopy-based navigation systems can be used for internal fixation of intracapsular femoral neck fractures, with the object of optimizing positioning of the implant and reducing radiation exposure. With this technique, the virtual position and direction of a reamer can be simultaneously superimposed on anteroposterior (AP) and axial (AX) fluoroscopic images. However, surgeons have to rely on the accuracy of these systems, because the only intraoperative feedback on the true position of the reamer is the projection of a virtual reamer superimposed on two fluoroscopic images. The objective of this study was to evaluate the accuracy of the displayed position of the virtual reamer in relation to the true position of the instrument when using a fluoroscopy-based navigation system (medivision, Oberdorf, Switzerland). Secondary to this, the accuracy of the drill-channel measuring tool of the system was analyzed. The study was performed on 20 sawbones. To evaluate the position of the virtual reamer, an 8-mm Perspex bar was inserted in predefined drill channels in each sawbone. AP and AX fluoroscopic images of the sawbones with the Perspex bar were loaded into the workstation. The Perspex bar was then removed and exchanged for a navigated dynamic hip screw (DHS) reamer. The position of the Perspex bar in the images represented the true position of the reamer. Subsequently, the difference between the position of the virtual reamer and the Perspex bar was measured with a dedicated computer program. Drill-channel lengths measured with the system were compared with measurements obtained with a digital ruler. The mean difference in position of the Perspex bar and reamer at a predefined point was 0.90 mm (range: 0.00-3.21 mm) in 360 images. The mean difference in length measurements between the medivision system and the digital ruler was 1.00 mm (p = 0.01, SD =1.33). Reaming and measuring the screw channel of a DHS with a medivision fluoroscopy based navigation system can be performed with an acceptable error margin. PMID- 12582981 TI - Registration accuracy and practicability of laser-directed surface matching. AB - The aim of our study was to evaluate the registration accuracy and practicability of a new laser registration technique in comparison to marker registration. From January to August 2001, 36 patients (23 male, 13 female) with brain lesions were operated with navigational guidance. Thirty-five patients were registered by paired-point registration. In 16 patients, a second registration was carried out using a special laser pointer for surface matching (z-touch trade mark, BrainLab, Heimstetten, Germany). Accuracy was evaluated by touching seven anatomic landmarks and a target fiducial with the nonsterile pointer. The distance from the virtual pointer tip to these points was determined on the monitor display (600% zoom). Laser registration is fully retrospective and allows registrations when no markers are applied. z-touch trade mark registration is more sophisticated and time-consuming than marker-based registration (registration time for z-touch trade mark = 7.4 +/- 3.7 min; for markers = 4.1 +/- 1.7 min; p < 0.005). Marker registration proved to be more accurate than z-touch trade mark registration with regard to localization of anatomical landmarks and target fiducials (precision with z-touch trade mark = 2.77 +/- 1.64 mm; with markers = 1.31 +/- 0.87 mm; p < 0.01). Although the registration error was increased and preparation time prolonged with the z-touch trade mark technique, it proved to be a valuable option, especially in children. PMID- 12582982 TI - Assessment of hepatic motion secondary to respiration for computer assisted interventions. AB - This article reviews the published efforts to characterize hepatic motion secondary to respiration, with the specific goal of defining the limitations and potential applications of image-guided systems in percutaneous liver interventions (computer assisted interventions). Hepatic motion and deformation due to respiration remain obstacles to applying stereotactic localization techniques to the liver. Respiratory-associated hepatic motion is complex. Nine studies using diagnostic imaging or modeling are reviewed, and their findings are tabulated herein. The significant variations in their findings are discussed, including cranio-caudal translation, anterior-posterior and lateral translation, movement secondary to tissue deformation, and motion with respect to surrounding tissue. Techniques for correcting for hepatic respiratory motion are then described, including gating techniques, modeling approaches, real-time liver tracking, and magnetic tracking technology. PMID- 12582985 TI - On the role of chiral catalysts in the alkenyl zirconocene/zinc addition to aldehydes: a study of ligand loading and asymmetric amplification. AB - Unusual nonlinear asymmetric amplification and chiral ligand loading effects were discovered for the use of catalytic quantities of chiral aminoalcohols in the in situ hydrozirconation-transmetalation-aldehyde addition processes. While the stereochemically most efficient aminothiol ligands demonstrated mechanistically conventional reaction parameters in excellent agreement with Kagan's ML(2) system, the asymmetric induction in the presence of a chiral aminoalcohol was found to vary greatly with loading and %ee of the ligand. Aminothiols remain the ligands of choice for the highly enantioselective formation of allylic alcohols and provide experimentally more predictable reaction variables. However, new, optimized conditions lead to a synthetically useful product %ee using the readily available and scalable aminoalcohol 2a. PMID- 12582987 TI - Lipase regio- and stereoselectivities toward three enantiomeric pairs of didecanoyl-deoxyamino-O methyl glycerol: a kinetic study by the monomolecular film technique. AB - A kinetic study was carried out on the regio- and stereoselectivities of 12 lipases of animal and microbial origin. For this purpose, monomolecular films consisting of three pairs of enantiomers (didecanoyl-deoxyamino-O methyl glycerol, DDG) containing a single hydrolyzable decanoyl ester bond and two lipase-resistant groups were spread at the air-water interface. Each of the lipases tested displayed a particular type of behavior, on the basis of which they were classified in two groups, depending on their ability to hydrolyze the sn-2 position. From the qualitative point of view, the sn-2 preference measured on triacylglycerides and DDG were in good agreement. The inductive chemical effect might explain why a greater level of hydrolytic activity was observed with the diglycerides than with DDG. With most of the lipases tested, it was observed that the enantiomeric pair having two distal acyl chains was more clearly differentiated stereochemically than the two homologous pairs with two adjacent acyl chains. This finding is consistent with the hypothesis that during the chiral recognition process two of the three attachment points may be the external (distal) hydrophobic chains, which is in line with the hypothesis of a tuning fork conformation of a triglyceride in the lipase active site. PMID- 12582986 TI - Reduction of flobufen in pig hepatocytes: effect of pig breed (domestic, wild) and castration. AB - Knowledge of the biotransformation processes of veterinary drugs and food supplements in food-producing animals is increasingly important. Residual levels of parent compounds or their metabolites in food of animal origin may differ with the breed, breeding conditions, and gender of animals. The nonsteroidal antiinflammatory drug flobufen, 4-(2',4'-difluorobiphenyl-4-yl)-2-methyl-4 oxobutanoic acid (racemic or its individual enantiomers) was used as a model to evaluate differences in activity, stereoselectivity, and stereospecificity of reductases in primary cultures of hepatocytes from intact male or castrated male domestic pigs (Sus scrofa domestica) or male wild pig (Sus scrofa scrofa). Time dependent consumption of flobufen enantiomers and formation of dihydroflobufen (DHF) diastereoisomers as their principal metabolites in hepatocytes were measured using chiral HPLC. Flobufen reduction in hepatocytes from all three experimental groups of animals was stereoselective ((+)-R-flobufen was predominantly metabolized) and stereospecific (2R;4S-DHF and 2S;4S-DHF diastereoisomers were preferentially formed). Flobufen reductases activity in male domestic pigs was 30 times higher compared to castrated pigs. Flobufen reductases activity was similar in domestic and wild pigs. The stereospecificity and stereoselectivity of DHF production did not significantly differ with breed or castration of animal. Chiral inversion of flobufen enantiomers was also studied and differences between castrated and intact male pigs were seen. PMID- 12582988 TI - Geometrical approach to central molecular chirality: a chirality selection rule. AB - Chirality is of primary importance in many areas of chemistry and has been extensively investigated since its discovery. We introduce here a description of central chirality for tetrahedral molecules using a geometrical approach based on complex numbers. According to this representation, for a molecule having n chiral centers it is possible to define an "index of chirality chi." Consequently, a "chirality selection rule" has been derived which allows the characterization of a molecule as achiral, enantiomer, or diastereoisomer. PMID- 12582989 TI - Evaluation of absolute configuration of naphthylphenyl-substituted oligosilanes by CD exciton chirality method. AB - Absolute configurations of methylnaphthylphenyl-substituted oligosilanes, MeNpPhSi*SiMeR(1)R(2) [2 (R(1), R(2)=Me), 3 (R(1)=Me, R(2)=Ph), 4 (R(1), R(2)=Ph), and 5 (R(1)=Me, R(2)=SiMe(3))] were predicted by circular dichroism (CD) exciton chirality method. The sigma-pi conjugation effect of oligosilylene units (sigma-linkage) with pi-electron systems caused an intense red-shift of (1)L(a,Ph) transition band of the oligosilanes as shown in UV/VIS and made it possible to observe clear CD exciton chirality between the two aromatic chromophores on chiral silicon atom. PMID- 12582990 TI - Role of latent heat in chiral symmetry breaking transition in the crystallization of 1,1'-binaphthyl. AB - The influence of latent heat dissipated by the crystallization of 1,1'-binaphthyl in its supercooled molten state on the chiral symmetry breaking transition was investigated. Temperature change in the crystallization system was monitored by infrared thermocamera. Temperature rise due to the dissipation of latent heat in the growing front of polycrystalline aggregate was about 2 degrees C in an unstirred crystallization system. The melting point of racemic mixture and racemic compound of 1,1'-binaphthyl is 145 degrees C and 158 degrees C, respectively. The latent heat generated by the crystallization could thus change the crystallization behavior when the initial temperature of the melt was slightly lower than 145 degrees C. The temperature change in both unstirred and stirred crystallization systems was monitored. In the stirred crystallization system, even in the case when the initial temperature of the melt was about 2 degrees C lower than 145 degrees C, the temperature rose by about 4 degrees C immediately after the onset of crystallization. This indicates that the role of stirring as the critical parameter for the chiral symmetry breaking transition is not only to clone the chiral crystals but also to enhance the dissipation of latent heat due to secondary nucleation. PMID- 12582991 TI - Conformational preferences in diastereomeric(5S)-methyl-3-(o-aryl)-2,4 oxazolidinediones. AB - The thermally interconvertible diastereomers of the (5S)-methyl-3-(o-aryl)-2,4 oxazolidinediones were synthesized and their conformers studied by (1)H NMR and HPLC. The barriers to rotation about the N-C(aryl) bond were found to be very much solvent dependent. For the o-fluoro oxazolidinedione, difference in barriers to rotation in deuterated methanol and deuterated chloroform amounted to 34 kJ/mol. ortho-Bromo substitution increased the barrier to rotation up to 100 kJ/mol in ethanol, which enabled the analytical separation of the diastereomers and observation of the thermodynamic enrichment of the S-P conformer by HPLC. In CDCl(3) by (1)H NMR, on the other hand, a barrier of only 89 kJ/mol was determined. The S-M and S-P conformers of the diastereomers of o-methyl, alpha naphthyl and o-iodo derivatives have been assigned by NOESY experiments and the kinetic and thermodynamic constants for the interconversion between the S-M and S P conformers were determined. PMID- 12582992 TI - Detection by circular dichroism of conformational transitions in pH and thermosensitive copolymers based on N-isopropylacrylamide and N-methacryloyl-L leucine. AB - Circular dichroism (CD) spectra in the region of 210-250 nm allow visualization of intrachain phase transition of pH- and thermosensitive polyelectrolytes. Indeed, in 0.001 M citrate and acetate buffers, at pH 4.0-5.5, aqueous solutions of a poly(N-isopropylacrylamide-co-N-methacryloyl-L-leucine) (NIPAAm-MALEU) copolymer containing 90.9 mol% of NIPAAm residues exhibit a well-defined sigmoidal increase in the CD signal at 220 nm with increasing temperature. This phenomenon is suggestive of a highly cooperative transition which occurs at lower temperatures compared to that observed by cloud point measurements. The change in the CD signal is less sharp at higher pH, indicating varying cooperativity with pH. For pH 6.0 and higher, no such phenomena are observed. PMID- 12582993 TI - Chiral reagents for the determination of enantiomeric excess and absolute configuration using NMR spectroscopy. AB - Recent advances in the development of chiral derivatizing and solvating agents that facilitate the determination of enantiomeric excess and absolute configuration are reviewed. These include metal-containing species, host-guest systems, donor-acceptor compounds, and liquid crystal discriminating agents. In the aggregate, these reagents can be used to analyze a wide range of compound classes. PMID- 12582994 TI - Modeling the chromatographic enantioseparation of aryl- and hetarylcarbinols on ULMO, a brush-type chiral stationary phase, by 3D-QSAR techniques. AB - The enantiomers of a series of chiral aryl- and hetaryl-carbinols were separated on a chiral stationary phase (ULMO) based on (S,S)-3,5-dinitrobenzoylated 1,2 diphenylethane-1,2-diamine. In all cases, the homochiral analogs of (S)-1 phenylethanol (mostly also S-enantiomers) were retained more. Log alpha values were modeled with the aid of the 3D-QSAR techniques CoMFA (comparative molecular field analysis) and CoMSIA (comparative molecular similarity indices analysis) as well as a descriptor based on normal mode eigenvalues (EVA). Partial least squares analysis with two (CoMSIA) or three (CoMFA, EVA) latent variables on a set of 22 training analytes gives cross-validated correlation coefficients q(2) = 0.85-0.91 and conventional correlation coefficients r(2) = 0.94-0.99. The quantitative structure-enantioselective retention relationships derived thereby were used to predict the separation factors of the test set, containing also hetaryl-carbinols, e.g., furan and thiophene analogs, with good accuracy. PMID- 12582995 TI - Enantiomeric discrimination of pyrethroic acid esters on polysaccharide derived chiral stationary phases. AB - Enantiomeric separation of pyrethroic acid methyl and ethyl esters was examined on cellulose-based chiral stationary phases (CSPs): chiralcel OD (cellulose tris(3,5-dimethylphenyl carbamate)) and chiralcel OF (cellulose tris(4 chlorophenyl carbamate)). The good resolution of pyrethroic acid esters was achieved on chiralcel OD and OF. Separation factors ranged from 1.19-5.12 for Chiralcel OD and 1.00-1.59 for chiralcel OF. Hexane/2-propanol (100:0.15, v/v %) was used as the eluent. The resolution capability of CSPs was greater chiralcel OD than chiralcel OF in the case of the pyrethroic acid esters. The flow rate was 0.8 ml/min and detection was set at 230 nm. The results of the chromatographic data and molecular mechanics suggest that steric effect was a major factor in the enantioseparation. Furthermore, the hydrogen bond between analytes and CSP played an important role in the chiral recognition. PMID- 12582998 TI - Determination of fluoxetine and norfluoxetine in rat plasma by HPLC with pre column derivatization and fluorescence detection. AB - Both fluoxetine (FLX) and its N-demethylated metabolite, norfluoxetine (NFLX), have been reported to be potent serotonin-reuptake inhibitors. A sensitive and reliable method that allows simultaneous quantification of their plasma levels would be valuable and was developed in this work. The procedure included extraction of FLX and NFLX from plasma, fluorescence derivatization with 4-(N chloroformylmethyl-N-methyl) amino-7-nitro-2,1,3-benzoxadiazole (NBD-COCl), separation of the derivatives on an octadecylsilica column with acetonitrile water (55:45,v/v) as mobile phase and fluorescence detection with emission at 537 nm and excitation at 478 nm. The calibration curves were linear for FLX and NFLX concentration over the range of 10-1000 nM (r = 0.9992 and r = 0.9997) and the limits of quantitation were 10 nM in 100 micro L of plasma. Precision of intra- and inter-day RSD of less than 12% and accuracy of intra- and inter-day RE within -6.0-13% were achieved. The method described was applied to analysis of the plasma samples from rats treated with FLX hydrochloride and to the pharmacokinetic study. PMID- 12582999 TI - Synthesis and analysis of aminochromes by HPLC-photodiode array. Adrenochrome evaluation in rat blood. AB - The catecholamine oxidation process induces cardiotoxicity and neurotoxicity. Catecholamines can oxidize to aminochromes through autoxidation or by enzymatic or non-enzymatic catalysis. Although some toxic effects seem to be related to the formation of aminochromes there is still scarce information concerning the identification and evaluation of these compounds in in vivo models. In this study five catecholamines were oxidized to their respective aminochromes: adrenaline/adrenochrome; noradrenaline/noradrenochrome; dopa/dopachrome; dopamine/dopaminochrome; and isoproterenol/isoprenochrome. The evaluation of the catecholamines oxidation profile was performed by HPLC with photodiode array detection and using either enzymatic (tyrosinase) or non-enzymatic [Ag(2)O, CuSO(4), NaIO(4) and K(3)Fe(CN)(6)] catalytic systems. The NaIO(4) was found to be the most efficient oxidant of catecholamines. An isocratic reverse-phase HPLC method was developed to analyse each pair of catecholamine-aminochrome. The analytical system was then applied to the detection of adrenochrome in rat blood at 490 nm. Thus, adrenochrome was administered i.p. to rats and its concentration in whole blood was monitored after 5, 15 and 25 min. Blood treatment for adrenochrome evaluation consists of an acidification for protein precipitation followed by a rapid neutralization. The results showed a rapid decrease of adrenochrome concentration in blood after its administration. The adrenochrome present in blood was characterized by UV and tandem mass spectrometry. PMID- 12583000 TI - SPE/RIA vs LC/MS for measurement of low levels of budesonide in plasma. AB - A radioimmunoassay is described that measures budesonide in plasma after solid phase extraction (SPE/RIA) of the analyte. The performance of the assay was compared with that of a selective LC/MS method. The limit of quantitation of budesonide determined for the LC/MS and SPE/RIA assay was 50 pg/mL and 120 pg/mL, respectively. Based on quality control samples, a higher variability was observed for the SPE/RIA (CV between 4.5 and 23.0%) than for the LC/MS method (CV between 7.5 and 12.5%). Plasma samples obtained from healthy volunteers after administration of budesonide rectal foam were assayed by both methods. In a subset of samples, these results were compared with those measured by direct RIA to evaluate the selectivity of two assays. About two times higher budesonide levels were measured with the direct RIA (lacking the extraction step), presumably because of cross-reactivity with budesonide metabolites, indicating that the extraction step in SPE/RIA is necessary for selectivity. Both SPE/RIA and LC/MS methods were found to be selective, sensitive and suitable for pharmacokinetic studies. Results obtained from the two methods were compared with a number of statistical methods. Ratios of results obtained for the clinical samples were close to 1 (ratio LC-MS/ SPE/RIA = 0.98 +/- 0.27). Linear regression indicated a slope of 1.17 +/- 0.0378. The concordance correlation (r = 0.91) indicated that the agreement between both methods was fair while the Bland-Altman plot indicated that the agreement was less pronounced at higher concentrations (1 3 ng/mL). In summary, the results confirm that the SPE/RIA is an alternative to HPLC/MS and that among the statistical methods tested the concordance correlation analysis was judged to be the most informative test to assess the comparability of two methods. PMID- 12583001 TI - Quantitative determination of saquinavir from Caco-2 cell monolayers by HPLC-UV. High performance liquid chromatography. AB - The validation and quantitative determination of the protease inhibitor, saquinavir, from confluent Caco-2 monolayers and from aqueous solution is reported. The high performance liquid chromatographic method consisted of an Ultramex 5 C(8) reverse-phase column (250 x 4.6 mm i.d.) and a mobile phase of acetonitrile:water:triethylamine (55:44:1, v/v/v, pH 6.5). Samples were analyzed using an ultraviolet detector at 238 nm, and diltiazem hydrochloride (66 micro g/mL) was used as an internal standard. A linear response over a broad concentration range (0.4-8.0 micro g/mL, r(2) = 0.997) was obtained. The limit of detection and quantitation was set at 0.14 and 0.4 micro g/mL, respectively. Over a 4 day period, the intra-day and inter-day precision ranged from 1 to 7% with a mean of 4%, and from 1 to 2% with a mean of 1.5%, respectively. Bench-top and storage stability of saquinavir was found to be satisfactory. The permeability of saquinavir through Caco-2 monolayers was estimated using this assay. PMID- 12583002 TI - Altered expression of insulins I and II and their mRNAs in the islets of Langerhans in dexamethasone-induced diabetic rats. AB - Rats have two isomeric insulins (insulins I and II). There have been no reports on the expression of the isomeric insulins in glucocorticoid-induced diabetic rats. To clarify the relation of the expression of each insulin and its mRNAs in dexamethasone-induced diabetic rats, the amounts of the isomeric insulins and mRNAs in the islets of Langerhans were determined in vivo and in vitro. A sensitive and selective HPLC-fluorescence determination method for the isomeric insulins and a newly developed real-time quantitative RT-PCR method for their mRNAs were used. There was a greater reduction of insulin II than insulin I in the islets of Langerhans in dexamethasone-induced diabetic rats. This alteration may be caused by a disproportionate expression of the respective mRNA for the isomeric insulins that resulted from the direct effect of dexamethasone. In addition, continuous hyperglycemia may also suppress the expression of the insulin II mRNA. The overall effects of dexamethasone and hyperglycemia may cause a greater reduction of insulin II than insulin I in the dexamethasone-induced diabetic rat. Conversely, an elevated ratio of insulin I to II in the islets could suggest a diabetic condition. PMID- 12583003 TI - A simple method for nicardipine hydrochloride quantification in plasma using solid-phase extraction and reversed-phase high-performance liquid chromatography. AB - A simple and sensitive reversed-phase liquid chromatography method was developed and validated for the determination of nicardipine hydrochloride (NC) in rabbit plasma. Nicardipine hydrochloride and nimodipine, used as internal standard, were initially extracted from plasma by a rapid solid-phase extraction using C(18) cartridges. After extraction, nicardipine hydrochloride was separated by HPLC on a C(18) column and quantified by ultraviolet detection at 254 nm. A mixture of acetonitrile-0.02 M sodium phosphate buffer-methanol (45:40:15) with 0.2% of triethylamine of pH of 6.1 was used as mobile phase. The mean (+/-SD) extraction efficiency of NC was 77.56 +/- 5.4, 84.23 +/- 4.32 and 83.94 +/- 3.87% for drug concentrations of 5, 25 and 100 ng/mL, respectively. The method proved to be linear in the range of 5-100 ng/mL with a regression coefficient of 0.9993. The relative standard deviations of intra- and inter-day analysis for NC in plasma were 3.26-6.52% (n = 5) and 4.71-9.38% (n = 5), respectively. The differences of the mean value measured from the concentration prepared, expressed in percentages (bias percentage), were only - 5.2, 0.4 and 0.8% at NC 5, 25 and 50 ng/mL, which confirmed the accuracy of the method. The analytical technique was used to determine NC plasma concentration after drug oral administration to rabbits. The results inferred that NC is rapidly absorbed in rabbits and has a short half-life (t(1/2) = 1.34 h). PMID- 12583004 TI - Laser-induced fluorescence as a powerful detection tool for capillary electrophoretic analysis of heparin/heparan sulfate disaccharides. AB - In quest for high sensitivities necessary for determining the disaccharide composition of heparin/heparan sulfate present in trace amounts in biologic samples, an ultrahighly sensitive capillary electrophoresis (CE) method using laser-induced fluorescence (LIF) detection was developed. Heparin/heparan sulfate derived Delta-disaccharides were derivatized with the fluorophore 2-aminoacridone and resolved by a reversed-polarity CE method. Estimation of the limit of detection in concentration term and limit of quantitation showed that LIF detection of AMAC-derivatives of Delta-disaccharides resulted in 27-744 times higher sensitivity as compared to those detected by UV at 255 nm. These data suggest that CE-LIF is a powerful tool to quantify minute amounts of heparin/heparan sulfate disaccharides. PMID- 12583005 TI - Capillary electrophoresis and enzyme solid phase assay for examining the purity of a synthetic heparin proteoglycan-like conjugate and identifying binding to basic fibroblast growth factor. AB - Interaction of basic fibroblast growth factor (bFGF) with heparin/heparan sulfate proteoglycans protects the growth factor against proteolytic degradation and is essential for its cellular activity. Although the structural requirements of heparin and heparan sulfate for the high-affinity binding to bFGF have been extensively examined, studies on intact heparin proteoglycans are limited. In this report, the purity and the binding ability of a heparin proteoglycan-like molecule-the heparin-bovine serum albumin (heparin-BSA) conjugate-was examined using capillary zone electrophoresis (CZE). Furthermore, the affinity of bFGF binding to the heparin-BSA conjugate was studied using an enzyme solid-phase assay. Chondroitin sulfate, dermatan sulfate, hyaluronan, heparan sulfate and variously sulfated disaccharides derived from heparin and heparan sulfate were also studied for their ability to compete with the binding of bFGF to heparin. Heparin-BSA conjugate was synthesized by reductive amination and, following precipitation with 1.5 vols of ethanol-sodium acetate, it was obtained free of contaminating heparin. Heparin-BSA-bFGF conjugate was obtained following incubation of heparin-BSA with bFGF for 2 h at 37 degrees C. Intact heparin, heparin-BSA and heparin-BSA-bFGF conjugates were completely resolved by CZE using 50 mM phosphate, pH 3.5, as operating buffer, reversed polarity (30 kV) and detection at 232 nm. Competitive solid phase assay showed that, among the glycosaminoglycans tested, heparin exhibits the highest affinity binding to bFGF (IC(50) = 6.4 nM). Heparan sulfate showed a lower affinity as compared with that of heparin, whereas all other glycosaminoglycans and heparin/heparan sulfate derived disaccharides tested showed minute effects. The developed CZE method is rapid and accurate and can be easily used to identify bFGF-interacting heparin preparations of biopharmaceutical importance. PMID- 12583006 TI - Validation and application of a high-performance liquid chromatography/tandem mass spectrometry assay for sumatriptan in human plasma. AB - A sensitive and convenient high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) assay is described for the (5-HT(lB/lD)) receptor agonist sumatriptan in human plasma. Sumatriptan was recovered from plasma (81.8 +/- 6.8%) by liquid-liquid extraction. The mobile phase flow rate was 0.3 mL/min and consisted of methanol:water:formic acid (90:10:0.1, v/v/v). The analytical column (4.6 x 100 mm) was packed with Partisil C(8) (5 micro m). The standard curve was linear from 0.7 to 70.4 ng/mL (r(2) > 0.99). The lower limit of quantitation was 0.7 ng/mL. The assay was specific, accurate (percentage deviation from nominal concentrations were <15%), precise and reproducible (within- and between-day coefficients of variation <10.3%). Sumatriptan in plasma was stable over three freeze/thaw cycles and at room temperature for one day. The utility of the assay was demonstrated by following sumatriptan plasma concentrations in two healthy subjects for 8-12 h following a single 20 mg intranasal dose. PMID- 12583007 TI - Specific determination of unbound oxacillin in protein solution with cefoperazone by high-performance frontal analysis with chemiluminescence detection. AB - Unbound oxacillin concentrations in human serum albumin (HSA) solutions in the presence or absence of cefoperazone were determined using high-performance frontal analysis coupled with chemiluminescence detection (HPFA-CL). The HPFA was performed on an ISRP column with 67 mM potassium phosphate buffer of pH 7.4 and ionic strength of 0.17 as the mobile phase. The luminol-H(2)O(2)-Co(2+) system was employed in the chemiluminescence detection. The detection was highly specific for oxacillin in the presence of cefoperazone. Although both drugs in HSA solutions co-eluted in the same region in HPFA, cefoperazone did not interfere with the determination of unbound concentration of oxacillin. In the solution of 100 microM HSA and 11.33 micro M oxacillin the bound percentage of oxacillin to HSA was estimated as 80.5%. Addition of 30.98 micro M cefoperazone into the HSA-equilibrated solution produced little effect on the protein binding of oxacillin. In the presence of 154.9 micro M cefoperazone, however, the bound percentage of oxacillin was significantly reduced. This specific method could be applied to the investigation of drug-drug interaction in protein binding. PMID- 12583008 TI - Pharmacokinetics of a new sustained-release formulation of theophylline sodium glycerinate in healthy subjects with a new asymmetric dosage regimen. AB - Pharmacokinetics of a new sustained-release formulation of theophylline sodium glycerinate in healthy subjects was studied. In this study, a new asymmetric dosage regimen was presented to achieve a better accordance with the chronotherapy of asthma. Each of 10 subjects was administered one tablet (equivalent to 0.1 g anhydrous theophylline) in the morning and four tablets in the evening for a consecutive 6 days and blood samples were collected at the predetermined time and analyzed by a validated HPLC method. This new regimen produced a steady and effective level of theophylline in plasma for the whole day, especially in the evening. A lower dose in the morning could reach the effective level (C(min)4.97 +/- 1.60 microg/mL and C(max)10.68 +/- 1.80 micro g/mL over the a.m. dosing interval) and a higher dose in the evening did not result in toxic levels but led to a reasonable concentration range (C(max)9.72 +/ 1.56 microg/mL over p.m. dosing interval), which could maintain a higher plasma theophylline concentration without the risk of serious adverse events and control asthmatic symptoms probably occurring during the night or early in the morning. The results suggested that the proposed asymmetric regimen was necessary, practicable and safe for twice daily sustained-release tablets of theophylline sodium glycerinate and also provides the basis for the clinical dosage regimen of other theophylline formulated products. PMID- 12583009 TI - Interaction of carbamazepine and promethazine in rabbits. AB - The interaction of carbamazepine and promethazine in rabbits has been investigated. The influence of this interaction on the processes of biotransformation in the liver was revealed. The drugs were administered as single oral doses (100 mg of each drug) as well as simultaneously with an interval of 15 min. The sequence of administration of the drugs was varied. The influence of promethazine on the pharmacokinetics of carbamazepine is expressed by: (a) strong suppression of carbamazepine's level in plasma and appearance of multiple peaks of carbamazepine; (b) suppression of biotransformation of carbamazepine into carbamazepine-10,11-epoxide at the initial stages and its increase in the intermediate stages. These data are explained by the active capture of carbamazepine by liver at its primary transferal through the liver and sufficient presystem elimination of carbamazepine in the presence of promethazine. The character of kinetic curves of promethazine varies substantially under the influence of carbamazepine. However, this change is not as strong as in case of carbamazepine. The concentration of promethazine in plasma varies slightly and multiple peaks are not observed. The rate of terminal elimination of promethazine varies and abrupt prolonged segments of elimination appear at the initial and terminal stages of the process in return. These data perhaps indicate the induction of biotransformation of promethazine in the presence of carbamazepine-an inductor of microsomal liver enzymes. The changes of kinetics of promethazine and carbamazepine by simultaneous administration as compared with their administration separately, as well as a comparative consideration of pharmacokinetics of promethazine and carbamazepine by simultaneous administration show the existence of competition in the elimination between these drugs and the periodic saturation of liver for their biotransformation. PMID- 12583010 TI - Nondestructive FTIR monitoring of leaf senescence and elicitin-induced changes in plant leaves. AB - The applicability of the FTIR attenuated total reflectance technique for in situ monitoring of plant physiological processes such as leaf senescence and aging has been examined. Difference spectra obtained by subtracting the spectrum of the young plant leaf from that of the older one revealed positive bands at 1650-1500 cm(-1), indicating a higher relative concentration of phenolics in the older leaves of both black cherry and sweet pepper bush leaves. Prolonged physiological stress of tobacco leaves exhibited a progressive time-dependent increase of the absorbance at around 3475 cm(-1), corresponding to hydroxyl functional groups. Absorption changes were also observed between 1650 and 1500 cm(-1), which are likely to correspond to phenolics. The characteristic changes of the FTIR absorbance spectra resulting from physiological and induced aging were detected also as a response to treatment with a recombinant alpha-elicitin, cinnamomin. This allowed the first quantification of the biological activity of a recombinant elicitin using a spectroscopic method. We suggest that FTIR spectroscopy provides important information about physiological events occurring in plant tissue in vivo, and it could be useful for the in situ characterization of the plant responsiveness to fungal toxins such as elicitins. PMID- 12583011 TI - 1H-NMR and 13C-NMR lipid profiles of human renal tissues. AB - Lipids from human renal tissues are studied by means of (1)H- and (13)C-NMR spectroscopy. The total lipid fractions obtained from healthy kidneys, malignant renal cell carcinomas, and benign oncocytomas are characterized and analyzed to elucidate the main differences between the functional and neoplastic tissues. In all cases the lipid components are well identified. The healthy kidney is characterized by high amounts of triglycerides and the presence of cholesterol in its free form. On the contrary, renal cell carcinomas contain high amounts of cholesterol that are almost completely esterified as oleate, suggesting an intracellular localization of the cholesteryl esters synthesis. Cholesteryl esters are considered markers of renal cell carcinomas, thus supporting recent theories that these compounds play a leading role in cell proliferation. Oncocytomas are particularly rich in phosphatidylcholine and, analogous to the healthy kidney, are completely lacking in cholesteryl esters. Healthy kidneys and oncocytomas appear to have other similarities if compared with renal cell carcinomas: a very high fatty acyl/cholesterol ratio, the presence of dolichols, and a higher grade of unsaturation. The (13)C data suggest a new method for the direct evaluation of the saturated/unsaturated fatty acyl ratio. PMID- 12583012 TI - Fluorescence spectral properties of cyanine dye labeled DNA near metallic silver particles. AB - Recent studies have demonstrated that silver metallic particles can increase the quantum yield and decrease the lifetimes of nearby fluorophores. These studies are extended to double stranded DNA oligomers labeled with N,N'-(dipropyl) tetramethylindocarbocyanine (Cy3) or N,N-(dipropyl)-tetramethylindodicarbocyanine (Cy5). The proximity to silver particles increases the apparent quantum yields and decreases the lifetimes of the double helical DNA 23-mer labeled individually with Cy3 or Cy5. The decreased lifetimes are accompanied by apparently increased photostability of the labeled oligomers near silver particles. Because of spatial averaging across the sample these results are likely to significantly underestimate the effects of silver particles on labeled DNA localized at an optimal distance from the metallic surface. These results suggest that DNA arrays fabricated on substrates with silver particles can display increased sensitivity and photostability in the analysis of gene expression. PMID- 12583013 TI - Distance dependency of exciton coupled circular dichroism using turn and helical peptide spacers. AB - Porphyrins are promising chromophores for the investigation of the still unexplored area of 3-dimensional structural studies of proteins by using the exciton coupled circular dichroism (CD) method. The synthesis, conformational characterization by FTIR absorption and (1)H-NMR, and CD properties are described for a model bis-porphyrin system based on homooligo-[L-(alphaMe)Val](n) peptides as rigid spacers. In particular, the coupled CD phenomenon is experimentally detected, the intensity of which is modulated by the interchromophoric distance. These results extend and integrate those already reported with steroid, dimeric steroid, and brevetoxin bridges. PMID- 12583014 TI - Study of protein-protein interactions by fluorescence of tryptophan analogs: application to immunoglobulin G binding domain of streptococcal protein G. AB - The protein-protein interaction system often contains many fluorophores that may significantly interfere with the quantitative determination of the binding abilities. To solve this perplexing problem, we biosynthetically incorporated the two tryptophan analogs, 5-hydroxytryptophan and 7-azatryptophan, into the immunoglobulin G (IgG) binding domain of streptococcal protein G (PGBD). The exclusive excitation and novel fluorescence changes in both the intensity and anisotropy are beneficial to reporting the details of the interactions between PGBD and the IgG fragments and enable assessment of the binding abilities. The dissociation constants are estimated to be 0.28 microM for the binding of human Fc and 8.0 microM for mouse Fc. The results clearly demonstrate that labeling of tryptophan analogs has very little effect on the binding abilities and is broadly applicable to quantitatively studying protein-protein interactions in a whole biomolecular complex. PMID- 12583016 TI - [System design and implementation of a distributed PACS]. AB - It is inevitable for our country to take the concept of informationized hospital in order to improve diagnosis quality and reduce administrative overhead. However we can not follow the way that has been taken in developed countries because of the national conditions of China. So we have to take some effective strategies to implement our high performance PACS. Based on our concrete experience gained from the development of a PACS in Shanghai, this paper presents a distributed architecture of PACS and describes the design detail of several PACS core components. PMID- 12583015 TI - Molecular structures and interactions of repetitive peptides based on wheat glutenin subunits depend on chain length. AB - Synthetic and recombinant peptide models of the central repetitive domain of the high molecular weight subunits of wheat glutenin with different numbers of the consensus repeat motifs PGQGQQ + GYYPTSLQQ (21, 45, 110, and 203 residues long) and a recombinant 58,000-Da relative molecular mass (M(r) 58,000) repetitive peptide from a single subunit (1Dx5) are studied using Fourier transform IR spectroscopy. The spectra of the dry peptides are very similar; at low water contents (<76% relative humidity) there is an increase in beta-sheet structures in all peptides. However, on further hydration the content of beta sheets decrease and more beta turns are observed. The changes during the second step of hydration are very marked in the 21 and 45 residue peptides, but they are less apparent in the longer perfect repeat peptides. In the 110 and 203 residue peptides hydration results in increased contents of intermolecular beta-sheets and less beta-turn formation. In contrast, the beta-turn content of the M(r) 58,000 peptide increases during the second hydration step. The decreased extent of structure changes with increasing chain length indicates that cumulative intermolecular interactions, in particular hydrogen bonds, are an important factor in determining the structures in the solid state. The regularity of the perfect repeat sequences in the 21, 45, 110, and 203 residue peptides may favor the formation of larger stretches of intermolecular beta sheets. In contrast, the M(r) 58,000 peptide contains imperfect repeats (in common with native glutenin subunits), which may limit its ability to form intermolecular beta sheets. PMID- 12583017 TI - [A 3D brachytherapy treatment planning system]. AB - Brachytherapy dose prescription and treatment playing lag behind the state-of-the art for external beam therapy. We have developed a computer program on a SGI Indigo2 graphics workstation to plan the loading or stereotactic implantation of radioactive seeds. The treatment planning system, using data from the pre treatment diagnostic CT scan or MRI scan, contours target areas on sequential images creating a 3-dimensional target volume, and is based on a dose algorithm of TG-43 formalism. The treatment isodose contour is generated at each scanningplane, reconstructed sagittal and coronal views, and overlaid on the corresponding target contour for 3-dimensional coverage visualization. Volume dose histograms are generated for the target and neighboring normal structures. We also provide two orthogonal digitally reconstructed projected images identifying virtual source positions superimposed on for fluoroscopy guided loading applicators and sources. PMID- 12583018 TI - [A new telemedicine system]. AB - A new telemedicine system and its application are introduced in this paper. Many new technologies, such as multiteleconference, multibind of MODEM and ISP, are adopted, which greatly improve the performance of the system. PMID- 12583019 TI - [Design of the system for detecting impedance cardiography based on windows plantform]. AB - In this paper is introduced systematically the design of the system for detecting impedance cardiography (ICG) based on Windows plantform. The three major ICG indexes of 30 students have been detected by the designed system in the clinical application, and conform with the results from the reference. On the other hand, compared with the system based on DOS plantform, the system has higher detection speed, and becomes more convenient, more flexible by the introduction of Mouse. PMID- 12583020 TI - [An analysis of S/N ratio of the phonocardiogram exercise test]. AB - Phonocardiogram exercise testing (PCGET) is a method through applying a phonocardiograph to evaluate cardiac contractility and the cardiac reserve. In order to test the certainty of PCGET method, a study on its S/N ratio was performed. Thirty volunteers performed PCGET. The average value of baseline amplitudes after exercise with respect to the average value of baseline amplitudes before exercise is 1.150; The average value of number of time of the S1 amplitude after exercise with respect to the S1 amplitudes before exercise is 10.57. The results suggest that PCGET has increased S/N ratio; introcardiac noise, noise from transmission of heart/thorax acoustic system, respiratory noise, muscle noise and ambient noise could not interfere in the application of PCGET. PCGET might be a noninvasive, convenient, and inexpensive technique to quantitatively evaluate cardiac reserve for abnormal or normal persons. PMID- 12583021 TI - [Wavelet packet-based ECG compression]. AB - A wavelet packet-based algorithm for the compression of ECG is presented in the paper. The wavclet packet algorithm, which synthesizes the efficiency and the flexibility of the wavelet pacheket, generates significantly lower data rates with less computational effort. PMID- 12583022 TI - [Research and development of the computer aided design for face lifting and image processing system]. AB - A medical image Processing system, in which the three dimensional image technology is applied to make the surgery planned quantitatively and imaged, is introduced in this paper. In the system, a computer image processing technology finds a use for medical face-lifting, which allows the patient to preview the effectiveness of the face-lifting. PMID- 12583023 TI - [The study of X & gamma rays' co-character imaging and its value]. AB - This paper discusses the X & gamma rays' identical and similar components in clinical imaging and physical Characters. A research has been made on the possibilities of their Co-character imaging and the economic benefits and significance are also discussed here. PMID- 12583024 TI - [Electric field effect of cell calcium]. AB - The mechanism of biological effects of extremely-low-frequency electromagnetic field involve induced changes of Ca2+ transport through plasma membrane ion channels. In this paper we discusses the effects of externally applied Vi = 30 sin 100 pi t(mV) on the induced Ca2+ flux of HL-60 leukemia cells. After signal excitation or sham excitation the response of the intracellular calcium levels changes not only with the different concentrations of the activator but also with the activated states of cells. PMID- 12583025 TI - [Applications of micromechatronics in minimally invasive surgery]. AB - With the development of Micro Electro Mechanical System(MEMS), it's application in the fields of biomedical engineering becomes a vital consideration. This paper describes the advantages of Minimally invasive Surgery (MIS) and its applications. A novel medical endoscope driven by micro robot is also introduced. PMID- 12583026 TI - [The principle and application technology of plasma sterilization]. AB - Plasma sterilization is a new sterilization technology. The advantages of this method are low temperature, time-saving and non-poison. The good sterilization effectiveness has been confirmed through experiments. PMID- 12583027 TI - [Real-time techniques in monitoring secretion activity in cells]. AB - Chemical communication for neurons and endocrine cells depends on their secretion activity. Development of variety of biophysical techniques have greatly promoted the understanding of the mechanism of cellular secretion. Here we will introduce three recently developed biophysical techniques which have been frequently used to monitor secretion activity in real-time. PMID- 12583028 TI - [The realigation of hospital computer network system based on Intranet/Internet]. AB - An introduction to the realization of the Intranet/Internet hospital computer network system is given here in the paper. PMID- 12583029 TI - [Generation of the electromagnetic interference and its eliminating method]. AB - This paper describes the generating conditions of electronic fields, magnetic fields and RF interference and the potential effects and hagards they may bring about on the medical devices. In addition, the basic principles and the methods of eliminating interferences are presented here. It may be very useful to the location, overall arrangement, the operation and the maintenance of the medical devices. PMID- 12583030 TI - [Image displacement elimination method in tomosynthesis]. AB - Computerized Tomography needs projected images from all directions, tomosynthesis doesn't need the information from all directions, but its reconstruction image is indistinct. In this paper, we introduce Frequency Domain Deconvolution Imaging(FDDI) which proposed recently. FDDI overcomes shortcoming of tomosynthesis, but FDDI's calculation is large, and its zero frequency will lose. We propose a new method-image displacement elimination method(IDEM), it overcome shortcomings of FDDI and can solve convolution equation set in space domain quickly. PMID- 12583031 TI - [A multichannel infrared telemetry system for physiological signal]. AB - This paper introduces the multichannel infrared telemetry system for temperature, respiration and ECG measurements. The experimental results are given at the end of the paper. This infrared telemetry system can be used in the environment with high electromagnetic interference, such as the operating room with electrosurgical unit and become an important monitor device. PMID- 12583032 TI - [The implement of remote medical treatment by Internet & RAS]. AB - This paper introduces a new method by taking full advantage of Internet and RAS technology, which can implement remote medical treatment perfectly. This method has been proved to be practical and will facilitate chinese hospital digital and information construction. The features and the advantages of of both technologies are introduced. Their designs & implement, and the prospect of this proposed method in china are discussed in details. PMID- 12583033 TI - [The development and clinical application of a multipurpose neuroendoscope]. AB - The multipurpose rigid neuroendoscope MGE-4 has been developed with the application of the self-focussing lens technology. We have been using it to examine and treat 10 neurosurgical patients who suffered from tumours, hydrocephalus, intracerebral haematomas of head injury and hypertensive cerebral haemorrhage. The short-term effect was satisfactory and there was no operative complications. It is considered that Model MGE-4 with the accessories can be used safely for the brain functional subcortical lesion or the deep-seated cystic lesions and haematomas. PMID- 12583034 TI - [The development of intelligent multi-function instrument of treatment and rehabilitation for disablement of limb]. AB - Intelligent multi-function limb treatment and rehabilitation instrument based on IBM-PC is introduced in this paper. It is mainly used for treatment and rehabilitation in the disablement of limb. The range speed and training time can be controlled and monitored. The instrument has the advantages of high reliability, easy operation and compact structure. It is a new kind of intelligent limb rehabilitation instrument. PMID- 12583035 TI - [The development of the ionization chamber and the researching of its characteristics for automatic exposure controlling in the X-ray diagnosis machine]. AB - An ionization chamber for automatic exposure controlling used in X ray diagnosis machine was developped, and its characteristics were measured and researched. The electrodes of measuring field were made of carbon, such couldn't leave it's imaging on the film. In order to increase the ionization efficiency and decrease the thickness and electric capacity between electrodes, the chamber was constructed in multilayers. The chamber we developed can be fixed in different x ray machines convenientlly, there is a fine linearity between it's signal output and the exposures, the beam quality response is uniform and output-timing response is also good. PMID- 12583036 TI - [The design and implement of PACS components in digital imaging apparatus]. AB - The domestic application of PACS will surely be developed in the future, but it will still take a few years. This paper proposes a tradeoff scheme: Attaching to digital imaging apparatus the PACS components, which mainly contain the Auto archiving unit, the Communication unit supporting WAN as well as LAN and the Converting Interface unit to the hospital PACS. In this way, a single digital imaging apparatus can archive and transmit data independently, also can merge with the hospital PACS conveniently in the future. In this paper, both the design and the implement are discussed in detail. PMID- 12583037 TI - [The restoration of the "double peak" in OCT images based on inverse Wiener]. AB - It has been found that the high reflectivity film of the reference mirror in OCT may cause a socalled "double peak effect" in the result images. To eliminate this a restoration algorithm is proposed here. The method is based on the mathematical modals of the point-spread function of OCT with consideration of double peak effect. The power spectrum of the noise and signal are estimated by the average smooth periodogram method. An inverse wiener filter algorithm, which avoids the illness condition in common inverse filter, gives the final restored images. The algorithm is also capable of the image degrading recovery due to the aberration and the band-limited spectrum. PMID- 12583038 TI - [A new adaptive filtering method for micro-particle image processing]. AB - Based on the local gray level histogram of macro-particle image which is more special than other images, a new adaptive noise filtering method for macro particle image processing is developed and described in this paper. It is made up of median filter and special order statistic (OS) filters (alpha-mean filter and inverse alpha-mean filter). This adaptive filter not only has satisfactory performance in both of noise-removal and edge-preservation of macro-particle image, but also is easy and quick to implement. An example is presented in this paper. PMID- 12583039 TI - [New parameter of liver ulitasonic tissue characterization--multi-fractal spectrum]. AB - Based on the previous research about the feature of liver ultrasonic images, this paper presents a new analysis method of multifractal spectrum alpha-f(alpha). Using a series of partition Hausdorff dimension f(q) this paper describes the sophisticated fractal characteristics of the ultrasonic images in the different section of the growing process and the multi-fractal spectrum alpha-f(alpha) acts as the new parameter of the ultrasonic issue characterization. Using the neural network it has catalogued and recognized the tissue diseases, caring out ultrasonic tissue characterization. Apparently, this method has practical significance. PMID- 12583040 TI - [EMG signal analysis based on fuggy and neural network]. AB - Recognizing methods of EMG signal based on the fuzzy and neural network, applying to a gamma model network is trained for EMG signal of artificial-limbs, compared with assemble-category analysis. The results demonstrated that identification rate is being the better. PMID- 12583041 TI - [The study on edge detection method of medical color image]. AB - Considering the specificity of the medical color image, in this paper a medical color image is decomposed as three color orthogonal feature: I1, I2 and I3 in K-L transformation method, and Kirsch algorithm is used to detect the edge of the monochrome image I1 including rich information. Finally, both clarity edge and rich detail of color backdisplay image are gained, if we adopt I2 and I3 to compensate I1. PMID- 12583042 TI - [The development of a two-channel impedance pneumography]. AB - To obtain respiratory signals with impedance method at two measuring frequencies, a high-performance two-channel impedance pneumography was implemented and the software including acquisiting data, showing waves, storing datas were developed. Human experiment was done and the results were also presented. PMID- 12583043 TI - [The designing of a software for an automated biochemistry analyzer]. AB - The structure, functions, user's interface, data storage and administration of a software system for an automated biochemistry analyzer are described. The methods to display the Chinese character menus and on-line help information are discussed in detail. PMID- 12583044 TI - [Temperature measurement and control in prostate therapy apparatus]. AB - The importance and difficulties of the temperature measurement and control in Prostate therapy apparatus are discussses. Plans of the temperature measurement and control, and the characters of the clinical applications are compared. Finally the method of calculating the temperature distribution with few measurement signals and the advantages of using the adaptive generalized predictive control are expounded. PMID- 12583045 TI - [A new type of 1-channel electrocardiograph drive circuit]. AB - This article introduces the driving circuit and damping circuit for the printing driver of the feedback printing system of the XD-106 1-channel electrocardiography. PMID- 12583046 TI - [Effect on new electrode technology with implantable motor prostheses for stimulating peripheral nerves]. PMID- 12583047 TI - [Quality assurance and control in SRS 200X--knife]. PMID- 12583048 TI - [Fractal dimension analysis of the surface EMG]. AB - Fractal analysis is a new method for biomedical signal processing. Present research on surface EMG and its parameterization method were introduced in this paper. Experiments were performed on three conditions: (1) different muscle contraction degree, (2) different static load, (3) different motion pattern. Problems and prospects in engineering application were also discussed here. PMID- 12583049 TI - [Design of blood cell analyzers by using stationary suspending laser light scattering method]. AB - The paper analyses the technology development of automatic blood cell analyzers, put forward the new method of blood cell counting and classification by stationary suspending laser light scattering. The new automatic blood cell analyser is designed by using this method. The design idea of the light route and data acquisition system is stressly provided in the paper. The calibration is performed by using the standard blood cell particulate. PMID- 12583050 TI - [A review on registration and fusion of medical images]. AB - Based on the criterion whether extracting the features of the image or not, the content and research status of medical image registration are discussed in this paper. PMID- 12583051 TI - [Study on measuring method of pulse oxygen saturation in clinical monitor]. AB - In this paper, a measuring system of pulse oxygen saturation is presented, which is based on the Lambert-beer Principle, using the relative intensities at two wavelengths in the red and infrared ranges. In order to meet the special demand of continuous clinical physiological monitoring, two circuits and appropriate algorithms are designed, one circuit, a feedback circuit controlled by micro controller, automatically regulates the base line of signal, another circuit is a controlled integral circuit which automatically regulates the amplitude of waveforms. As the result, the drift of base line in this system, especially the effects caused by the physiological differences of human bodies, which will deteriorate the system, have been reduced. The system is satisfactory to clinical monitoring. PMID- 12583052 TI - [The nail fold micro-circulation images edges detection based on wavelet transform]. AB - In this paper, we discuss the application of wavelet transform to edges detection, classification and description. Then, according to the peculiarity of the nail fold micro-circulation images, dyadic wavelet transform is successfully applied to detect the edges and the Mallat algorithm is used in the wavelet decomposition. PMID- 12583053 TI - [A new digital filter method for eliminating 50Hz interference from the ECG]. AB - The article discusses advantages and disadvantages of several conventional digital filter methods. Then, based on Levkov method, the author proposed a new filter method. We use these methods to remove 50 Hz interference from more than 50 persons' ECG signals. The results show that this new method is the best, and it can satisfy the real time requirement of digital ECG machine. PMID- 12583054 TI - [The problems and solutions of the leakage current checking in medical electrical equipments]. AB - It is an important safety checking item to check leakage current of medical electric equipments. In the national standard, there are definite and strict stipulations for the leakage current-checking and the checkers. But this important checking is often misrepresented with general current measurements. So the checking is not up to the standard and the results are not dependable. In this paper, the special demands for the checking and the key technique of the checkers have been expounded in detail. PMID- 12583055 TI - [A heart function measuring and analyzing instrument based on single-chip microcomputer]. AB - An Introduction a measuring and analyzing instrument, based on the single-chip microcomputer, which provides sample gathering, processing, controlling, adjusting, keyboard and printing. All informations are provided and displayed in Chinese. PMID- 12583056 TI - [Medical shape memory alloy]. PMID- 12583057 TI - [The study of the control system of new generation EECP]. AB - A new algorithm for the inflation and deflation timing of the Enhanced External Counterpulsation (EECP) have been developed, A pentium PC and 16 bit microcontroller with parallel interrupt communication were used for developing a new kind of EECP system. The 16 bit microcontroller was used to collect and pre process the ECG and EECP signals. The pre-processing data is transmitted to PC by the parallel communication interface. According to the new algorithm, PC sent out the sequential control signals and monitor the state of EECP. It is necessary to synchronize the time of aortic valve closure and diastolic augmentation during EECP, under the condition of the patients' safety, the effect of therapy have been extremely improved. PMID- 12583058 TI - [The design of a multi-parameter patient monitor]. AB - This paper describes the software and hardware design of a multi-parameter patient monitor which is able to measure ECG, heart rate, blood pressure, breath rate, body temperature and oxygen saturation non-invasively. The instrument uses embedded industry control module as its main controller and 9" CRT as its display. Several OEM modules are utilized in the design, such as ECG module, blood pressure module and oximeter module. This design strategy has the features of reliable performance, easy maintenance, and short developing period. PMID- 12583059 TI - [3-channels data acquisition system based on single-chip-microcomputer used in the measurement of coronary sinus blood flow]. AB - The coronary sinus blood flow can be figured out, which based on the principle of thermodilution, so long as gets the temperature of blood, indicator and mixture of blood and indicator respectively. This system is a smart slave module with single-chip-microcomputer. The structure and principles of hardware and the flow chart of software are described in detail. PMID- 12583060 TI - [The design of a reflection sensor of NIRS tissue oximetry]. AB - A design of the reflection light-electric Sensor suited to monitoring the statues of tissue blood oxygen on the surface of body is described. The designed sensors have been used too to detect blood oxygen statues of brain and muscle respectively, and many useful informations have been acquired Therefore The above mentioned design is reasonalle and Feasikle. PMID- 12583061 TI - [The development of an intelligent vision testing instrument]. AB - The paper introduces an intelligent vision testing instrument, which works on automatic method or manual method. The examinee inputs his or her judgement about the direction of the letter "E" with remote control. The system examines and judges the examinee's judgement, and then prints the testing report. The vision testing range is from logarithm vision 3.0 to 5.3. The distance between examinee and optometer can be measured automatically by the system with infrared and ultrasonic testing functions and a good accuracy. PMID- 12583062 TI - [The development of a hemorrhologity observating and measuring instrument]. AB - A new instrument for observing blood cell state and the process of variation on hemorrheology has been developed. There are three ways of shear rate: setting by hand, automatically setting and simulating pulsative state. It can provide different variable flow fields. The shear rate can be set in the range from 0.1 to 1000 s-1. It needs only a little blood. It can be easyly and rapidely operated. it has clear images. The hemorrheology state of blood cells can be directly perceived through thd instrument by eyes. The display and measurement and record of blood cell's hemorrheology state can be finished in real time dynamically and simultaneously. PMID- 12583063 TI - [Development and clinical application of dental spot weld & heat-treat tri-uses device]. AB - A spot weld & heating treatment tri-uses device based on modern mechanical electric technique, was developed for dental spot weld, stainless steel orthodontic wire heating-reversion and Ni-Ti orthodontic wire heating-overload metamorphism. The device will be very helpful instrument in orthodontic clinic. The paper not only described the operating methods of the device, but also discussed the Performance-improved principles of stainless steel orthodontic wire and the principles of heating overload metamorphism of Ni-Ti orthodontic wire. PMID- 12583064 TI - [The development of pressure-controlled cryobio-cooling system]. AB - In this paper, a new cooling system controlled by the pressure of LN2 container is introduced, the controlling method of this device is discussed too. PMID- 12583065 TI - [Development of the automatic dental X-ray film processor]. AB - This paper introduces a multiple-point detecting technique of the density of dental X-ray films. With the infrared ray multiple-point detecting technique, a single-chip microcomputer control system is used to analyze the effectiveness of the film-developing in real time in order to achieve a good image. Based on the new technology, We designed the intelligent automatic dental X-ray film processing. PMID- 12583066 TI - [The development of model DYC-I multi-purpose device for lumbar vertebra puncture]. AB - A multi-purpose device for lumbar Vertebra puncture, which is ablete obtain cerebrospinal fluid (csf) and to determine the intracranial pressure, is presented here. This device has the functions of avoiding blindness of puncture, measuring pressure hermetically, liquid crystal display and controlling the flow of CSF. PMID- 12583067 TI - [A new design of bileaflet heart valve to eliminate the blood flow disturbance]. AB - The major problems of current clinically--used bileaflet heart valves are thromboembolism and hemorrhage related to anticoagulants. These bileaflet heart valves are considered as being unsatisfactory, because in the open position, two leaflets divide the ring orifice of the valve into three regions. In this obstructive orifice, the high turbulent shear stresses and the region of flow separation and stagnation all give rise to the occurrence of thrombosis. In our design of bileaflet heart valve, in the open position, no structural component or leaflet divides the ring orifice of the valve. Our bileaflet heart valve has the best possibilities for creating more natural blood flow Pattern which could decrease or eliminate the blood flow disturbance so as to reduce the risk of thrombosis and the needs for anticoagulation therapy. PMID- 12583068 TI - [The extracting of left ventricular contour based on adaptive directional filter]. AB - This paper proposes the adaptive directional filter, which is exploited for the extracting of left ventricular(LV) contour. This algorithm could produce the exact ridge point by applying directional filter with smoothing operator in the angiography images damaged by noise the Edge Directional Vector as the result of this filter is efficient for predicting the edge ahead, while the filter parameters is adaptive. This proposed method provides the more precise and more automatic--extracting LV contour only with fewer interactions. The experimental results show that this proposed algorithm could improve the accuracy and reproducibility, and reduce the computational consumption remarkably. PMID- 12583069 TI - [Research on applications of micromachine technology in medical engineering]. AB - This paper presents the basic concept of micromachines. The development aim and Key technology of medical micromachine Research are discussed here too. Besides, this Paper introduces some typical application research of micromachines, which may have actual reference Value for the future development of medical engineering. PMID- 12583070 TI - [Study of dental model testing tool based on robot theory]. AB - A new three dimensional testing and analysing system of dental model is discussed It is designed based on the motion theory of robots. The system is capable of not only measuring the three dimensional sizes of dental models, but also saving and outputing the tested data. The construction of the system is briefly introduced here. PMID- 12583071 TI - [The method of wavelet transformation used for extracting the characteristic of the exercise ECG]. AB - This article introduces the basic rationale of wavelet transformation and the algorithm of typical dyadic wavelet transformation with mother wavelet of the first derivative of a Gaussian smoothing function, this wavelet transformation is used to recognize the characteristic parameters of the exercise ECG, it is concluded that this method make significant effects on restricting base line drifting and lessening noisy component in the exercise ECG, it provides a new way to do further research on characteristic recognition of the exercise ECG in the exercise test. PMID- 12583072 TI - [Study on producing medical oxygen by pressure swing adsorption from air]. AB - In this paper, some technical subjects such as absorbent selection, process development, multistage filters, were studied, and this will help to apply the technique in hospitals of our country. PMID- 12583073 TI - [The application of modularity design in the pulse oximeter]. AB - This paper presents a thought of modularity design and application in our pulse oximeter and its hardware and software composition of the function board, the application can improve the reliability of the instrument greatly and make it easy to expand its function. PMID- 12583074 TI - [Single-chip computer controlled acupoint nerve stimulator]. AB - This article introduces the technical Characteristic and principles of the single chip computer controlled electrical acupoint nerve stimulator. Some clinical practices of this instrument are also introduced. PMID- 12583075 TI - [The development of a RF electrical pole catheter for heart ablation]. AB - This article describes a RF electrical pole catheter for heart ablation. There is a device in the catheter, that can adjust the bending of the catheter tip on which there are a pair of electrical poles that is able to record and map the electrical potential of the heart, and to transfer 50 W electrical energy for focus ablution of heart disease. PMID- 12583076 TI - [The application of wavelet transform in medical image processing]. PMID- 12583077 TI - [The principles and apparatus of pulmonary function testing]. PMID- 12583078 TI - [The comparison between approximate entropy and complexity in the study of sleep EEG]. AB - The Paper introduces two algorithms as a complexity measure. One is the algorithm defined by Kaspar and Schuster, the other is a recently developed statistic quantifying regularity and complexity--Approximate entropy (ApEn). They have been applied for the study of sleep EEG. According to the laboratory application and comparison, the result shows that ApEn is a valued complexity measure. PMID- 12583079 TI - [Intensity-modulation radiotherapy using independent collimator: algorithm study]. AB - To investigate algorithms for independent collimator (IC) to modulate beam intensity distributions. Field-setting sequences were determined for three clinical intensity distributions with two forward algorithms. The delivery order of all fields in the sequences was optimized in order to minimize the overall jaw moving time. Suppose treatments were performed with Varian 600C, delivery time was calculated. The delivery time was between 5 min to 10 min, It was approximately inversely proportional to machine dose rate, and also related to algorithm, number of intensity levels and metrix element size. PMID- 12583080 TI - [The research of distributed HRV analysis system based on DCOM]. AB - This article introduces the meaning of Distributed Component Object Model(DCOM), which used network communication protocol to replace the local process communication to reach the aim of Distributed Computing, then it discusses the significance of HRV analysis, and finally it presents the system we developed. PMID- 12583081 TI - [Detection of ECG characteristic using wavelet transform]. AB - This paper discusses the characteristic detection of modulus maxima of the loavelet transform, based on the properties of the singularity of the signal and noise. Theoretical analysis and experimental results show That the method is fitting for ECG signal detection and positioning. PMID- 12583082 TI - [Quadrature head coil of open mark 2000 MRI system]. AB - By using Quadrature combining technology in Open Mark 2000 MRI system of An Ke Inc., we developed quadrature head coil which is better than normal solenoid coil by 20% in signal to noise ratio. PMID- 12583083 TI - [The testing methods of the medical ultrasonic linear array transducer]. AB - This paper explores the testing methods of the linear array transducer in regard to its integrality, Uniformity and independence. PMID- 12583084 TI - [The design and analysis of sensitivity & filter circuits in the electrocardiograph]. AB - This paper introduces a sensitivity selection circuit and a filter circuit in the electrocardiograph. PMID- 12583085 TI - [The development of a bilateral groove lengthener]. AB - A new series of bilateral groove external fixator are introduced in the article, which are mainly used to treat limb crispation. These apparatuses are composed of the groove frame threaded rod, hooked bolts, nuts and pins. They can be adjusted in several directions to correct the axis of limb in limb lengthening. The pins can be fixed on the anterior and posterior sides of the frame, with a stable angle structure. The devices are light and practical, with multiple functions, low price, simple and convenient operation. It can apply to all kinds of hospitals. PMID- 12583086 TI - [Electrical impedance tomography technology]. PMID- 12583087 TI - [Review and new development of QRS complexes detection methods]. PMID- 12583088 TI - [A general evaluation method of the hospital managing system software quality]. PMID- 12583089 TI - [A minitype acceleration circulation device]. AB - This devices uses the circular pulsating air pressure to apply the contracting and stretching actions to the patient's extremities section by section, for promoting the back flow of vein blood and lymph in the extremities. It has better functions in prevention and cure of extremities' edema and vein twist expansion in lower limbs that caused by various block of circulation as well as the forming of vein thrombus in the bedfast patients for a long time. PMID- 12583090 TI - [Detecting method of the clinical visual electrophysiological signals]. AB - According to the characteristics of anatomy and electrophysiology of vision, this paper describes the method of sampling along the visual path to obtain the integrated information of electrophysiology of vision. The developed system MET01 can detect and analyze EOG, ERG and VEP signals as a whole, and has a clinical value. PMID- 12583091 TI - [Multi-bed & multi-parameter central monitoring system based on TCP/IP protocol]. AB - Communication is one of the key problems to a central monitoring system. In this paper we put forward a central monitoring system using TCP/IP as the network protocol, Windows NT4.0 as the platform, forming a Intranet in a hospital. We also discussed the communication problem between the bed-side monitoring station and the central monitoring station in detail and then put forward a new protocol- Hospital Central Monitor Protocol (HCMP) based on TCP/IP to transfer monitoring data. It is easy to achieve tele-monitoring through the current communication subsystem. PMID- 12583092 TI - [A multiparametric monitoring network system based on windows operation system]. AB - This paper introduces the general structure and implementary method both in hardware and software of a monitoring network system, which is developed for monitoring physiological multi-parameter under Windows operation system and based on personal computer. The parameters monitored by the system include Electrocardiogram (ECG), Noninvasive Blood Pressure (NIBP), Oxygen Saturation (SpO2), Respiration (RESP) and Body Temperature. Monitoring data can be shared by internal hospital network. The system has the advantages of easy operation and flexible structure. It is a new kind of hospital monitoring equipment. PMID- 12583093 TI - [Design of blood-pressure parameter auto-acquisition circuit]. AB - This paper presents the realization and design of a kind of blood-pressure parameter auto-acquisition circuit. The auto-acquisition of blood-pressure parameter controlled by 89C2051 single chip microcomputer is accomplished by collecting and processing the driving signal of LCD. The circuit that is successfully applied in the home unit of telemedicine system has the simple and reliable properties. PMID- 12583094 TI - [The development of a multifunctional molecular electrophoretograms processing system]. AB - This paper presents to you the principles, composition and features of the multifunctional molecular electrophoretograms processing system. PMID- 12583096 TI - [The development of a original multifunctional peritoneal biopsy needle]. AB - A multifunctional peritoneal biopsy needle is introduced in this paper, which can be used for peritoneal biopsy, brush biopsy and routine, biochemical and cytological examination. The clinical applications show that it could raise markedly the diagnosis rate of ascites, and especially it has important clinic diagnosis value for tuberculous and cancer ascites, and primary peritonitis complicated by hepatocirrhosis. It is simple, safe and practical to operate this needle. PMID- 12583095 TI - [A new generation of pulmonary function measuring system]. AB - A system for measuring the pulmonary function was reported. The System consists of Fleisch flow transducer, a pressure transducer an amplifier, A/D converter, and a 486 computer. It can measure the indices of respiratory function under tidal breathing, the indices of tidal breathing flow volume loop and the dynamic indices of passive respiration. PMID- 12583098 TI - [Comparative study of ultrasound transducers in HIFU]. AB - The high intensity focused ultrasound (HIFU) surgery technique is an important application of medical ultrasound. As the ultrasound transducer is placed out of the body, HIFU technique belongs to non-invasive extracorporeal therapeutic technology. It is able to focus ultrasound energy deep into the body, forming destruction in pathologic change tissues and coagulating them without any damages to tissues out of the focus. The development of ultrasound transducer is the key technology of HIFU. PMID- 12583097 TI - [Virtual heart studies and its applications]. AB - This paper presents the development of virtual heart studies and its applications. PMID- 12583099 TI - [The stereotactic radiotherapy technology: current status and development]. AB - There in this paper is an introduction of the radiotherapy characteristics and its equipment. PMID- 12583100 TI - [The development of a SPECT processing system for reports with both graphics and text]. AB - A computer designed for writing reports is connected to the Ethernet of SPECT. In the System, the remote FTP registration and the coping of SPECT graphics are used to obtain, at low cost SPECT reports with both graphics and a text, which are printed by a color printer and could provide adequate medical informations for clinical needs. PMID- 12583101 TI - [Optimizing the lab reports of Helena electrophoresis analyser with HRGL]. AB - The Paper describes HRGL(Helena Report Generating Language) general structure in detail and then introduces a method to optimize the report outcome, and an example is given for illustration. PMID- 12583102 TI - [The development of a brain surgery path planning system]. AB - Based on the patient's CT/MRI Section images, the brain surgery path Planning system makes surgeon freely draw out the interesting regions through the use of feasible interactive drawing contour function. And by fast judgement, the system is able to give out the risky value of the path. Finally, the system would directly show the surgery path in 3-D display. PMID- 12583103 TI - [AM signals' digital demodulation and its application in physiological signals' remote transmission]. AB - Taking advantage of the characteristics of digital systems, a new digital method is given in this article to overcome the difficulties in demodulating AM signals using digital methods. Physiological signals can't be transmitted through telephone networks because they are full of ultra-low frequency components. However, if they are modulated (such as AM), they can be transmitted to the receiving end through telephone networks. Then phaselocked loops can be employed to trace the frequency of the signals' carriers so that we can demodulate and transmit them in the end. PMID- 12583104 TI - [A new method to improve performance of BAM in presence of noise]. AB - Algorithm of bidirectional associative memory (BAM) is depicted. BAM sum up all sample pattern with different weighting factor to recall the true pattern. The ability to recall pattern correctly is determined by how the sample pattern are weighted. Selecting the maximum of weighting factors by competition can get best performance of BAM in presence of noise. PMID- 12583105 TI - [Gastric mucosa blood-supply analyzer]. AB - A new medical device, using a gastroscope and the OMA technology, is intoduced in this paper. It is used to determine the blood-supply parameters (oxygen saturation, hemoglobin and onygenated hemoglobin) of gastric mucosa quickly and noninvasiuely. Besides, its testing principles and structure are described here. The clinical testing resuets obtained from various parts of the stomach and gastric ulcer show that they are very important and very useful informations for clinicd diagnosis. PMID- 12583106 TI - [The development of a double point sensor for the pulse]. AB - This article has introduced the research and design on a double point sensor for the comprehensive pulse. The difference of this sensor from the single point sensor is that it is able to obtain a mechanics parameter from the pulse signals of the traditional Chinese medicine. This would be a great help for the more detailed quantitative analysis of the pulse signals of the traditional Chinese medicine. It has also explored the contribution of the double point sensor to the clinical medicine. PMID- 12583107 TI - [Design of a data acquisition circuit for ventricular late potentials and electrocardiogram]. AB - This paper introduces a data acquisition circuit that is able to acquire ventricular late potentials and standard 12-lead electrocardiogram signals. This circuit adopts time interruption acquisition, it has the characteristics of programmed gain, isolated ground, low-noise, high-CMRR, collecting data for a long time. PMID- 12583108 TI - [Study on optical system of medical fiberscope]. AB - The optical imaging principles of medical fiberscoptic endoscopes and their design featuies are described with GIS-105 Fibreoptic Endoscope as an example in this paper. PMID- 12583109 TI - [The development of an automatic blood collector controlled by microcomputer]. AB - A new automatic blood collector developed by using microcomputer AT89C2051 is presented in this paper. Meanwhile its principles component block diagram and their functions' are introduced here. All clinical experiments have shown that it has the advantages of easy operation, reliable performance, lower price and so on. It is truly an ideal upgrade product of intravenous blood exampling. It would find various applications and would have a bright future. PMID- 12583110 TI - [Experimental safety study using rats and mice irradiated with the complex field cure instrument]. AB - The complex field cure instrument is a new medical instrument. The complex field is composed of several sorts of physical quantum fields. Mice and rats are continuously irradiated by the complex field for 20 days to 90 days. The result shows that WBC, Hb and PLT value of the animals irradiated with the instrument are more than the control group, and their biochemical index is normal. The internal organs of the experimental animals show no toxicity. The visceval coefficient (liver, spleen) of the experimental animals is more than the control group. The marrow micronucleas test is negative. The chromosome test show no distortion. All of the results have proved that the instrument produces no toxicity side effect. Application of the instrument is safe and reliable and it may increase immune function of the body. PMID- 12583111 TI - [The technique and progress of lithotriptors]. AB - This introduces the classifications, features and progress of lithotriptors, as well as compares applications among them. PMID- 12583112 TI - [The reasonable design of the combined spinal epidural needles (CSEN) in aneasthesia]. AB - This paper discusses the reasonable factors of parameters in CSEN design and has also raised some useful reasonable parameters concerned. PMID- 12583113 TI - [A hanger-needless infusion set and it's unique infusion tube]. AB - The design of an enclosed barrel with an interiorly compresed plunges is introduced in the paper. With a unique floating-bulb-design tube, the new infusion set makes liquid infusion easy at any height, in any direction, with no need of hanger and without any risk of theleast possibility of any air into blood vessels. It is suitable for prehospital rescue, battle field treatment and wounded Rersonnels' transpostation etc. The new infusion set can also be tailored or improved according to the different requirements of usess and clinical applications. PMID- 12583114 TI - [A biomechanical study of telescopic external fixator]. PMID- 12583115 TI - [Multiprocessor study and design for digital color ultrasonic medical imaging system]. AB - In this paper, the design of a multi-processor for digital color ultrasonic medical imaging system is discussed. It is a shared memory multiprocessor of MIMD architecture. The whole system is devided into several parts according to the complexity of the signal-processing, and each part is allocated a certain duties. Besides, the solving method of multiprocessor's bus conflicting is discussed and an architecture block diagram is given in the paper. PMID- 12583116 TI - [Image reconstruction based on tomosynthesis]. AB - As a reconstruction method earlier than CT, tomosynthesis has its characteristic. This paper first analyzes principles of tomosynthesis, through transformation of equation set of tomosynthesis, which describes relation between sectional image and projection image, and we have proved that tomosynthesis is the same as Algebraic Reconstruction Techniques essentially. Reconstruction of tomosynthesis is indeed the reconstruction of Algebraic Reconstruction Techniques which under limited angle. We may regard all directive projection information as constitution of two limited angle projection information. Finally we reconstruct sectional image by using conception of "row" in tomosynthesis. And computer simulation has proved our conclusion to be correct. PMID- 12583117 TI - [A surface EMG signal identification method based on short-time Fourier transform]. AB - A surface EMG signal identification method based on short-time fourier transform is presented in this paper. To fully utilize the nonstationary character of the EMG signal, short-time fourier transform is employed to get the signal's time frequency representation. Singular value decomposition is then used in the spectrogram to extract feature vector for pattern identification. Four types of movement of forearm, hand grasp, hand extension, wrist pronation and wrist supination can be identified from surface EMG signals. Experimental results show that it is a stable and efficient method for extracting features. PMID- 12583118 TI - [The development of event related-potential acquisiting system]. AB - A system for acquisiting event-related-potential signalsis presented, making use of random-sequences visial symbols. ATMEL AT89C51 controls Max 186 serial conversion and interface Max202 to serially send data to host computer for poct processing. PMID- 12583119 TI - [The development of an intelligent acoustic impedance middle ear analyzer]. AB - This thesis introduces the measuring principle, device composition and main functions of an intelligent acoustic impedance middle ear analyzer. As well the results of clinical trial are analyzed in this paper. As an objective diagnostic method, aural acoustic impedance measuring device holds a very important position in diagnosing ear diseases and hearing obstacles. PMID- 12583120 TI - [The development of a new version of multifunctional radioactive nuclide diagnostic instrument]. AB - A new kind of multifunctional radioactive nuclide diagnostic instrument is described here, which can make noninvasive measurements on the metabolic functions of five most important inner organs which are kidney, liver, heart and thyroid. The design of the hardware and software of the new instrument embodies many advanced advantages such as multifunctional integration, high accuracy and high reliability, computer controlled, automatic data-processing, easy to use and manage, and so on. Applications show that this is an advanced and efficient radioactive medical diagnostic instrument. PMID- 12583121 TI - [The development of a 64-channel epicardial potential mapping system]. AB - A 64-channel epicardial potential mapping system was developed in order to study the mechanics of arrhythmia such as atrial fibrillation and ventricular fibrillation, and instruct the procedure of detecting and eliminating abnormal rhythm in experiment or clinic. The system was consist of electrode, amplifier, A/D card, computer and output device. The system's software include signal acquisition module, signal preprocessing module, character-point detecting module, isochrone-map construction module, output display and print module. The system could be used to detect the activation path of atrial fibrillation and ventricular fibrillation. PMID- 12583122 TI - [The development of an oral biomechanical testing instrument]. AB - An oral biomechanical testing instrument, which is portable, powered with batteries and controlled by single chip microcomputer, was described. The instrument was characterized by its multichannel, high accuracy, low power dissipation, wide rage of force measurement and stable performance. It can be used for acquisiting, displaying and storing data. And it may be expected to be an ideal instrument for oral biomechanical measurements. PMID- 12583123 TI - [Cellular electrophysiology and patch clamp technique]. AB - The fundamentals of cellular electrophysiology and its basic method are described here. The simplyfied structure of the cell, the chemical constitutes of the membrane, and the mechanism of the bioelectric signal generation are included. The patch clamp is the fundamental methodology in modern electrophysiology research. It is closely related to the construction of the laboratory setup and the key technique in experimental procedure. PMID- 12583124 TI - [NZ-III management system of hospital equipment and appliances]. AB - This paper describes the design, composition and application of the management system of hospital equipment and appliances, The enforcement of the system has promoted the standardization of hospital equipment and appliances management, the accurate data quantization and establishing a rational procedure for decision making. PMID- 12583125 TI - [Study of helical CT low-contrast resolution and its influencing factors]. AB - This paper studies the helical CT low-contrast resolution and its influencing factors by experimental analysis. The CT low-contrast resolution increased with mA, kV, slice thickness, scan time and FOV, and decreased with matrix, pitch and thickness of the scanned objective. The reconstruction algorithm of high resolution results lower low-contrast resolution. Mostly helical image low contrast resolution is slightly lower than axial image. It's very important for us to understand CT low-contrast resolution and its influencing factors for the correct evaluation of the performance of CT scanner. PMID- 12583126 TI - [An automotic dialyser reuse device]. AB - This article introduces an automatic dialyser reuse device used to rinse and disinfect dialysers. The Forcing signal generated by automatic program controller is fed into an output circuit and amplified to operate the execute device to realize the mechanical operation for treating the dialyser and the blood lines according to the program. This automatic dialyser reuse operation devices is able to replace the manual operation completely and enhances the reuse operation performance. PMID- 12583127 TI - [The electrostatic protection of the medical devices & equipments]. AB - This paper describes the harms of the static electricity to medical devices & equipments and its protection. PMID- 12583128 TI - [The development of an intellectualized dynamic strain device for three dimensional cell culture]. AB - The design principles of an intellectualized dynamic strain device for three dimensional cell cultures based on microcomputer are mainly introduced here, which includes principles of hardware and software design of controlled unit, principle and structure of mechanical unit, and construction of three dimensional scaffold and culture unit. The main technical index and advantages of the device have been also discussed in the paper. PMID- 12583129 TI - [The dynamic measurements of human body's unbalance]. AB - A through study of the postural stability of 40 subjects has been conducted with a stabilometer and the computer system. The subjects stand on the testing platform for 10 seconds and meanwhile the sum of frequency spectrum is being analyzed. The most men subjects maintain a little sway well within timits during the trials. The testing conditions include open eyes, close eyes, 10 degrees, 20 degrees, 30 degrees of platform inclination, transverse vision tracking, longitudinal vision tracking and vectorial vision tracking, All testing results show that there are unbalance differences between different age groups under different conditions. PMID- 12583130 TI - [Ventricular wave identification in epicardial mapping system using wavelet transform]. AB - A novel method for the identification of ventricular wave based on wavelet transform is presented. First, "Multi-Scale Intersectional Search" is adopted for analysis of wavelet transform coefficients on some different scales, then adaptive detection is used on this basis. The results of practice show that it has a discrimination of 94.5%, and is obviously superior to any routime method. PMID- 12583131 TI - [A method for real-time pickup action signal]. AB - Generally, each single action is picked up from successive action EMG signal by hand. Then this colleced signal is discriminated after getting its features. But doing by this way is obviously difficult to meet requirements of the myoelectric artificial limb. So this paper presents an easy method from the myoelectric energy point of view. The action signal segments and no-action signal segments are picked up from colleced EMG signal. Thus it is convenient for the relevant follow-up processing, and avoids dealing with no-action signal data. Therefore it is of useful and practical significance for multifunction myoelectric control of prostheses. In this paper, it has been proved that this method is feasible by using it to pick up actual effective action signal. PMID- 12583132 TI - [The design of open magnet]. AB - The thesis introduces the whole designing process of open magnet used in MRI. In the process, we use a new algorithm that is called Integral Equation. According to this algorithm, we have made an optimized design and manufactured the main magnet. We also have made a comparison between the theoretical calculation results and the measued data. PMID- 12583133 TI - [Design of a infrared single-channel ECG telemetry system]. AB - A single-channel ECG telemetry system with infrared rays as the medium was designed. It can be used to monitor the subject's ECG in real-time. This system has functions of detecting abnormal ECG, alarming, dieplaying, storing and printing. It is a beneficial attempt in the clinical application of the infrared telemetry. PMID- 12583134 TI - [Development of multi-function ECG signal generator]. AB - This paper describes the development of a portable multi-function ECG signal generator, which is based on micro-controller. It uses technique of LCD screen, and realizes man-machine interaction by keyboard. In constructing and disposing data module of the ECG signal, Eigen-heartbeat Code mapping method gets ROM saved greatly. Therefore it can generate all kinds of user-defined ECG signal sequence with no extension of on-board memory chips. This system can also simulate kinds of ECG signals, which have various heart rates and symptoms. It can meet the needs of researching and maintenance of kinds of ECG instruments. PMID- 12583135 TI - [Nerve threshold value detector]. AB - This paper describes the design, the composition and the method of clinical application of the nerve threshold value detector, which offers a new way of surgical diagnosis and treatment for paralytics. PMID- 12583136 TI - [The development of a function-rehabilitation trainer for lower limb joints]. AB - We'd like to introduce to you an exercising trainer for the recovery of functions of lower limb joints. It is beneficial to early-stage rehabilitation of lower limb joints after the operation. The light-weighted, portabl trainer is easy to operate duc to its innovative structure and design. In addition, the speed, angle and time of exercise of this trainer is automatically adjusted to best fit your needs, making it an advanced trainer for recovery purpose. PMID- 12583137 TI - [The artificial liver's studies and progress]. AB - This describes new achievements and progress in artificial liver studies. PMID- 12583138 TI - [Stereotactic conformal radiotherapy technology]. AB - Body Stereotactic Conformal Radiotherapy technology was introduced in this paper. This discusses target localization, beam shape, treatment planning system and reproducibility. PMID- 12583139 TI - [Single channel and whole cell recordings using patch clamp technique]. AB - Action of ion channel on membrane is the key events of messenger transduction for excitable cells, which can be detected by the patch clamp technique. The developments of patch clamp technique have brought a revolution of life science research. The theory, work modes, single channel and whole cell recording techniques in single cells are described here in detail. PMID- 12583140 TI - [Physical parameter measurement and quality assurance of X-knife]. AB - The treatment of X-knife with linear accelerator, the results and the methods used in physical parameter measurement, are introduced in this paper. We also discusses some problems about quality control. It is proved that our results can be used in X-knife treatment. PMID- 12583141 TI - [The imaging principles and the structure of the low-dose digital radiographic device]. AB - This article introduced A new type of X-ray radiographic device, which uses MWPC as the low-dose detector with the machinery and computer to reconstruct the X-ray digital images. In this paper, the advantages and the disadvantages of the device are analysed, thus much improvement has been made to make it better and more efficient. PMID- 12583142 TI - [The research on applications of diagnostic software in single photon emission computed tomography]. AB - In this paper, the structure of software system, the principles of overlay programming and the running mechanism of the programs for the Single Photon Emission Computed Tomography(SPECT) made by French SOPHA MEDICAL Corp. are analyzed in detail. On this basis, a method of extending functions of the system is introduced too. All the results of the extension in software functions for organ volume determination and of the measurements for living animals have proved the correctness and reliability of the method. PMID- 12583143 TI - [Study on the permanent MRI technology]. AB - This paper describes the open permanent MRI technology, introduces the special technology such as main magnet field, gradient magnetic field coil, Transmitter/Receiver coils and so on, and points out that the installation and adjustment of the MRI are also very important for optimal operation and best image quality. PMID- 12583144 TI - [Control system for rotational gamma knife]. AB - The features and composition of Rotational Gamma Knife are mainly introduced in this paper. The open architecture double PLCs and motion controller are used to fit special requirements of the Rotational Gamma Knife and thus to ensure its high reliability. In the end, safety measures which should be taken for patients, doctors and equipments are discussed. PMID- 12583145 TI - [The development of a full-automated blood coagulation analyzer]. AB - This paper introduces the electronic principles and systematic construction functions of a full-automated blood coagulation analyzer, which can measure the blood coagulation rightly and effectively in the clinic. by using the scattered light turbidmetry. This analyzer overcomes some shortcomings existing in other analyzers such as high price, low resolution and no quality control function. PMID- 12583146 TI - [An integrated medical diagnostic X-ray unit controlled by microcomputer]. AB - This article introduces an integrated medical diagnostic X-ray unit controlled by microcomputer. It is a practical and low-priced spot-film radiography device, and it is applicable to conventional local control unit. If you select a set of X rayimage intensification, it can be remotely controlled. This device utilizes DC servo motor and fine precision feedback variable resistor and closed loop control so it has the advantage of fine static control and low mechanical noise. The whole radiographic process is automatically controlled by microcomputer, and can be operated easily. PMID- 12583147 TI - [Automated microbiology identification and antibiotic susceptibility analysis system]. AB - This article introduces the principles, characters and application prospect of Model 2000 Automated Microbiology Identification and Antibiotic Susceptibility Analysis System. PMID- 12583148 TI - [The development of a simulated experimental system on humoral integrated parameter tests controlled by a microcomputer]. AB - A simulated experimental system on humoral integrated parameter tests controlled by a microcomputer is developed for overcoming hard training in maintenance technical skill. This paper describes its structure, characteristics, functions and the prospects of its applications. PMID- 12583149 TI - [Study on digitized atlas of the human brain]. AB - Digitized Human Brain Atlas is an important tool for visualization of medical images and investigation of brain science. This paper describes its fundamentals, development, construction and applications in surgical planning, model-driven segmentation and neuroanatomy teaching. PMID- 12583150 TI - [Content-based medical image retrieval]. AB - At first, the concept of medical image retrieval based on contents is here illustrated. Then, features of medical image are analyzed. Among them, gray histogram method, texture method, shape method are discussed, and the compression method is emphasized. Finally, the challenges in medical image retrieval are presented before us in the paper. PMID- 12583151 TI - [Surgical sutures and their clinical applications]. AB - This article describes the development of surgical sutures and their clinical spplications. It is classified as absorbable and non-absorbable sutures. Besides it discusses the suture's tension strength and tissue reaction, points out the advantages and the shortcomings. PMID- 12583152 TI - [The development and application of HP-I closed thoracic cavity drainer]. AB - The closed thoracic cavity drainer is equipped with a general standard infusion bottle as the drainage bottle, which can be easily supplied by all hospitals. Therefore, the new drainer is a very practical, cheap medical device of easy operation. PMID- 12583153 TI - [FVR setting mode and the adjustment of SHIMADZU S/V 1250 X-ray machine]. AB - The tube current of the x-ray machine is one of the essential factors that decide the quality of a photogram taken by the machine. FVR setting mode introduced in this article takes advantage of the computer technology to adjust the tube current and thus to improve the quality of the photograph. PMID- 12583154 TI - [Basic assessment of trust level in medical apparatus and instruments sales systems by medical organization]. AB - Purchase of medical apparatus and instruments is a strict work of taking the patient as a consuming objection. To analyze the trust level in medical apparatus and instruments sales systems and to set up the method of trust level assessment before the purchase, is an important link of ensuring the reality of medical products and consumerism. According to the principle mentioned about, the present article analyses the basic content of assessing factors and their inter relationship, summarizes their expressions, studies and sets up the mathematical modal and it method. All these are of some value to medical organization in choosing factory owners and their products while the purchasing. PMID- 12583155 TI - [Diagnosis and integrative evaluation on soil fertility of three Chinese medicinal materials in GAP plots]. AB - The fertility of soil was diagnosed and integratively evaluated by modified Nemoro Index with eight indexes, i.e. pH, organic matter, total N, P, K and available N, P, K on three national Chinese medicinal materials GAP (Good Agricultural Practice) experimental plots of Pogostemon cablin, Citrus medica and Morinda officinalis in Guangdong. The results showed that the pH, organic matter and other major nutrient contents were low and imbalance. There are lots of nutritive disturbance factors. The level of soil integrative fertility in any of the plots was poor. PMID- 12583156 TI - [Effects of hormone proportion on different adventitious buds from explant of Ligusticum chuanxiong]. AB - The differential results of explants from Ligusticum chuanxiong in medica with different kinds and proportions of hormone were reported in this article. The result showed that the best medica for inducement of adventitious buds and growing into plantlets were MS + 6-BA 0.5 mg/L + NAA 0.5 mg/L and 1/2 MS + IAA 1.0 mg/L + NAA 0.5 mg/L respectively. PMID- 12583157 TI - [Preliminary survey on medicinal plants of Polygonum in Gansu Province]. AB - OBJECTIVE: To gain a clear idea on the resources of medicinal plants of Polygonum in Gansu. METHODS: Field investigation, specimen collection, taxonomic study, scientific names study and literature retrieval were carried out. RESULTS: The distribution, ecological environment and medicinal parts of 23 species and 7 varieties of Polygonum from Gansu have been clarified. CONCLUSION: The investigation may prove to be useful for the utility and further study of Polygonum in Gansu province. PMID- 12583158 TI - [Study on morphological and histological identification of herba pogostemonis]. AB - OBJECTIVE: To identify two types of Pogostemon cablin (Guang Huoxiang), Shipai Huoxiang and Hainan Huoxiang. METHOD: Morphological and microscopical methods. RESULTS: Long bag-like glandular hairs with one-cell head existing in the intercellular space are found in the cortex of stems, mesophyll and parenchyma of midrib. The glandular hairs with special large one-cell head and with a special short and small one-cell stalk, flat bag-like cuticle above secretory cell are caved in the forms of square, triangle, convex len and round-oids are also found in the surface of leaves and stems. The primary pholem fiber bundles with very thick walls are uncontinuous and arranged with a circle in the transverse section of stems; the perimedullary cells contain needle and prism crystals. Above mentioned characters are differed from other plants of Labiate family. The old stems are cylinderoid forms, branch angles are usually smaller than 60 degrees, the surface of upper, middle stems and young leaves with purple colour are unusual, the palisade ratio are less than 10(5.05 and 9.26 cultivated seperatedly in Guangzhou and Gaoyao), in Shipai Huoxiang. But in Hainan Huoxiang, the old stems are rectangular cylinder, branch angles are usually over 60 degrees, the surface of upper, middle stems and young leaves are commonly purple colour, the palisade ratio are over 10(14.35, 13.18, 13.84 and 14.33 cultivated separatedlly in Wuchuan, Suixi, Leizhou and Hainan). CONCLUSION: Above-mentioned characters may distinguish two types of Pogostemon cablin, Shipai Huoxiang and Hainan Houxiang. PMID- 12583159 TI - [Saponins with low sugar chain from the leaves of Panax notoginseng (Burk) F. H. Chen]. AB - Six saponins with low sugar chain were isolated from the leaves of Panax notoginseng (Burk) F. H. Chen. Their structures were elucidated as 20(R) ginsenoside Rh2(I), 20(R)-ginsenoside Rg3(II), ginsenoside Mc(III), ginsenoside F1(IV), ginsenoside Rh1(V) and daucosterol(VI) by spectroscopic analysis and comparison with authentic samples. Compounds I-IV were first isolated from the plant. PMID- 12583160 TI - [Studies on immunocompetent constituents of Patrinia scabra Bunge]. AB - Five compounds were isolated from the roots of Patrinia scabra Bunge separated and purified on sillica gel column chromatography. Their structures were elucidated on the physico-chemical properties and spectral data as lariciresinol(I), syringaresinol(II), scopoletin(III), quercetin(IV), ferulaic acid(V). All compounds were obtained from P. scabra for the first time. In vitro biological test of these compounds showed that syringaresinol was able to stimulate T and B lymphocyte proliferation. PMID- 12583161 TI - [Study on the effect of Rhizoma Curcuma Longa on gastrin receptor]. AB - By the method of receptor combination, effects of the active constituents in Rhizoma Curcuma Longa and Radix Curcumae were studied. The results showed that Rhizoma Curcuma Longa probably include the excitant or antagonist of gastrin receptor. And the inhibitory intensity was shown as followed: parts of ethyl acetate > parts of methanol > parts of ether > parts of volatile oil, root tuber > rhizome. PMID- 12583162 TI - [The study on the anti-oxidation effect of root of Mallotus apelta in the rat model of liver fibrosis]. AB - OBJECTIVE: To investigate the effect of root of Mallotus apelta in treatment of liver fibrosis and the effect of anti-oxidation in the rat model of liver fibrosis. METHODS: A rat model of liver fibrosis was induced with 40% CCl4 in peanut oil. A series of items were detected, which include ALT, AST, MDA, NO and hydroxyproline in the blood or liver tissues. RESULTS: The level of ALT, MDA, NO in serum were significantly decreased, the level of hydroxyproline was declined too, while the formation of liver fibrosis was successfully ameliorated. CONCLUSION: The root of Mallotus apelta could retain the progression of liver fibrosis, having a capacity of anti-oxidation. And the latter might be the reason of the former function. PMID- 12583163 TI - [Study on the scavenging action of polysaccharide of Usnea longissima to oxygen radical and its anti-lipi peroxidation effects]. AB - OBJECTIVE: To study the scavenging action of polysaccharide of Usnea longissima Ach. (PUS) to oxygen free radical and its inhibiting action on lipidperoxidation. METHODS: The scavenging action to superoxide anion free radical (O2-.) was tested by xanthine-xanthine oxidase system. The scavenging action on hydroxyl free racical (OH.) and anti-lipidperoxidation effects were tested by Fenton reaction. RESULTS: The scaverging action was represented by the content scavenging 50% of all the free radical(IC50). The IC50 for O2-. and OH. were 0.45 mg/ml and 1.57 mg/ml respectively. The lipidperoxidation induced by OH. was weakly inhibited by PUS. CONCLUSION: PUS can apparently scavenge oxygen free radical. But it can only weakly inhibit the lipid peroxidation of hepatocyte homogenate of mice which resulted form hydroxyl free radical. PMID- 12583164 TI - [Effects of Hovenia dulcis Thunb on blood sugar and hepatic glycogen in diabetic mice]. AB - OBJECTIVE: To observe the effects of Hovenia dulcis Thunb (HDT) on blood sugar and hepatic glycogen in diabetic mice. METHODS: The diabetes model was established by alloxan and treated with different doses of HDT for 7 days, while the positive control group was treated with glibenclamide. Then the blood sugar and hepatic glycogen were measured. RESULTS: The blood sugar of mice treated by HDT or glibenclamide significantly lower than diabetes model group. The hepatic glycogen in middle or low dose of HDT and glibenclamide groups was significantly increased. CONCLUSION: HDT has effect on lowing blood sugar and may become an effective anti-diabetic herb. PMID- 12583166 TI - [Extraction of ecdysterone from Stemmacantha uniflora]. AB - OBJECTIVE: To optimize the conditions for the extraction of ecdysterone from Stemmacantha uniflora. METHODS: Conditions for the extraction were studied by three different methods as guided by the content of ecdysterone and solid material that present in the extract. RESULTS: Methanol was the best solvent, water the second, ethanol was greatly effected by concentration. Ultrasonic extraction had a good efficiency with short time. The time of cool extraction was too long, its efficiency was inferior to the other methods. CONCLUSION: The best methods for the extraction of ecdysterone from Stemmacantha uniflora was methanol reflux extraction. PMID- 12583165 TI - [Effect on gastrointestinal functions of Polygonum paleaceum]. AB - OBJECTIVE: To study the effect on gastrointestinal functions of Polygonum paleaceum Wall. METHODS: Defecation reflex, gastric emptying, motility of the gastroinestinal tract of mice and isolated intestinal smooth muscle of rabbits were used. RESULTS: Polygonum paleaceum Wall could inhibit defecation reflex and depress the motility of the gastrointestinal tract, delay gastric emptying in mice. It also had analgesia action, but didn't affect rabbit intestinal contraction spontaneously or induced by acetylcholine (Ach) in vitro. CONCLUSION: It suggested that Polygonum paleaceum Wall possessed the action on gastrointestinal functions. PMID- 12583167 TI - [Study progress in Sinomenium acutum (Thunb.) Rehd. et Wils]. AB - This article reviewed the progress in the study of the pharmacognosy, chemical compositions, pharmacological actions and clinical practices of Sinomenium acutum (Thunb.) Rehd. et Wils. An expectation for the further development and utilization of this plant was put forward. PMID- 12583168 TI - [Study on nutritional characteristics of Pogostemon cablin]. AB - This paper studied the soil nutrient status, the nutrient contents of Pogostemon cablin nutritional organs, the relationship between soil available nutrients and nutritional elements contents of Pogostemon cablin, the nutritional elements contents of leaves and their proportions in different growth periods of the plant, the reuse of the nutritional elements of leaves in Pogostemon cablin base of Guangdong Institute of Chinese Medicine. The results provided scientific basis for the research on the fertilization and special fertilizers of Pogostemon cablin. PMID- 12583169 TI - [Effects of Atractylodes macrocephala seeds coated with SCF on field diseases]. AB - The field plot experiments were designed with L9(3(4)) orthogonal comparison to study the effects of Atractylodes macrocephala seeds coated with SCF on field diseases. The results showed that the treatment 2 was the best SCF to control field diseases. The treatment of seed-coating with thiophanate methyl reduced the percentage of field diseases significantly. PMID- 12583170 TI - [Study on the quality of Rhizome of Dioscorea persimilis]. AB - OBJECTIVE: To evaluate the quality of Rhizome of D. persimilis Prain et Burk. METHODS: Determined the contents of 6 qualitative indexes in samples of Rhizome of D. persimilis Prain et Burk that were cultivated in Yongning, Bobai, Beiliu, Guiping and Rongxian of Guangxi and in sample of Rhizome of Dioscorea opposita Thunb, that was cultivated in Wenxian, Henan province. RESULTS: The contents of water in them (%): 15.39, 18.52, 15.41, 13.96, 15.48, 15.74; the contents of protein in them (%): 7.06, 8.45, 9.20, 9.67, 8.63, 9.79; the contents of starch in them (%): 84.51, 86.51, 82.72, 83.65, 84.19, 79.13; the contents of water extracts in them (%): 6.19, 6.98, 8.95, 7.41, 6.70, 9.40; 90% EtOH extracts in them (%): 2.00, 2.04, 2.68, 2.20, 2.11, 3.53; the contents of 18 kinds of amino acids in them (%): 5.23, 6.31, 6.22, 6.78, 6.59, 6.83. CONCLUSION: The quality of Rhizome of D. persimilis Prain et Burk cultivated in Guiping or Beiliu is beter than that of Rhizome of D. persimilis Prain et Burk cultivated in Yongning. The quality of Rhizome of D. persimilis Prain et Burk cultivated in Rongxian or Bobai are between those of Rhizome of D. persimilis Prain et Burk cultivated in Guiping or Beiliu and Rhizome of D. persimilis Prain et Burk cultivated in Yonging. The qualities of Rhizome of D. persimilis Prain et Burk cultivated in Guiping or Beiliu are similar to that of Yam. PMID- 12583171 TI - [Interspecific and intraspecific comparison of valepotrates contents in three Valeriana plants]. AB - OBJECTIVE: To compare the contents of three valepotrates intraspecifically and intraspecifically in three medicinal Valeriana plants grown in different areas in China. METHODS: The method of RE-HPLC was adopted. RESULTS: The results showed that the contents of three valepotriates in Valeriana jatamansi Jones, V. officinalis L. and V. officinalis var. latifolia Miq. were different. Among them, that of Valeriana jatamansi Jones was the highest. The contents of three valepotrates in Valeriana jatamansi Jones varied significantly from different areas, samples collected from Longli of Guizhou province ranking the highest. The contents of three valepotriates in V. officnalis L. also varied significantly from different areas, samples from Liuba of Shaanxi province showing the highest. The results also showed that the contents of three valepotriates in underground part are all higher than that in aerial part. CONCLUSION: The interspecific and intraspecific differences of valepotriates in three Valeriana plans were obvious. PMID- 12583172 TI - [Description and histology identification of several algae of Sargassum sp]. AB - This paper reported the description characters and microscopical identification of seven kinds of algae of Sargassum sp., Sargassum pallidum (Tum.) C. Ag., S. fusiforme (Harv.) Setch., S. horneri (Tum.) C. Ag., S. hemiphyllum (Turh.) C. Ag., S. thunbergii (Mert.) O'Kuntze, S. polycystum C. Ag. and S. kjellmanianum Yendo. The results revealed that there were clear differences in the description characters and microscopical identification of the seven kinds of algae of Sargassum sp. These studies provided a scientific basis for distinguishing crude drug of algae, developing and making use of alga natural resources of Sargassum sp. PMID- 12583173 TI - [Triterpenes isolated from Poria cocos by derivazations]. AB - The separation of triterpene acids was achieved from Poria cocos (Schw.) Wolf by TLC after treatment with Ph2CNN or RX under mild condition followed by selective hydrolysis of ester groupings. Three triterpene acids, 3 beta-acetoxyl-16 alpha hydroxy-lanosta-8,24(31)-diene-21-oic acid (I), 3-oxo-16 alpha-hydroxy-lanosta 7,9(11), 24(31)-trien-21-oic acid (II) and 3 beta, 16 alpha-dihydroxylanosta 7,9(11), 24(31)-trien-21-oic acid (III) were isolated. PMID- 12583174 TI - [Studies on chemical constituents of Meconopsis quintuplinervia Regel]. AB - Seven flavonoids were isolated from Meconopsis quintuplinervia Regel for the first time, and their structures were elucidated as quercetin (I), dihydroquercetin (II), luteolin (III), chrysoeriol (IV), apigenin (V), huazhongilexone (VI) and hydnocarpin (VII) by spectroscopic methods including IR, EIMS, 1D- and 2D-NMR techniques. PMID- 12583175 TI - [Isolation, purification and structural characterization of active polysaccharides from the mycelium of Ganoderma lucidum]. AB - The active polysaccharides from the mycelium of Ganoderma lucidum were fractioned by DEAE-Sephadex and DEAE-Cellulose. Through repeated isolation and purification, two polysaccharides, GLMB0 and GLMB1, were finally obtained. Their structures were characterized by chemical and spectral methods. PMID- 12583176 TI - [Studies on the chemical constituents from Ampelopsis grossedentata]. AB - OBJECTIVE: To study the chemical constituents from Ampelopsis grossedentata (hand Mazz) W.T. Wang. METHODS: Chromatography and spectroscopic analysis were employed to isolate and elucidate the chemical constituents in the plant. RESULTS: Seven compounds were isolated and elucidated as ampelopsin (I), myricetin (II), myricitrin (III), gallic acid (IV), beta-sitosterol (V), stigmasterol (VI) and dihydroquercetin (VII). CONCLUSION: Among these compounds, IV, V, VI and VII were isolated from this plant for the first time, and compound VII was obtained from genus Ampelopsis for the first time. PMID- 12583177 TI - [Comparison studies on chemical constituents of essential oil from Ramulus Cinnamomi and Cortex Cinnamomi by GC-MS]. AB - Essential oil from Ramulus Cinnamomi (Cinnamomum cassia Presl) and Cortex Cinnamomi were obtained by steam distillation. The chemical components of the oil were examined and compared by means of gas chromatography-mass spectrometry. 54 of the separated constitutents were identified. 10 components of these were not observed in the oil from Ramulus Cinnamomi, and 16 components of these were not observed in the oil from Cortex Cinnamomi. PMID- 12583178 TI - [Protection of vascular endothelial cells from high glucose injury induced by quercetin]. AB - OBJECTIVE: To evaluate the effects of quercetin on the high glucose-injured vascular endothelial cells (VECs). METHODS: Experiments were divided into control group, injured group and quercetin group. The cultured VECs were injured by high glucose. The proliferation of VECs was assessed by MTT assay. The amount of NO and lipid peroxidation (monitored as maloraldehyde, MDA) of VECs were assessed by fluorometric assay. The amount of lactate dehydrogenase (LDH) was assessed by spectrophotometric method. RESULTS: 10(-2), 10(-3), 10(-4), 10(-5) mg/ml quercetin increased the proliferation of high glucose-injured VECs. 10(-2), 10( 3) mg/ml quercetin reduced LDH release and MDA production, increased NO release. CONCLUSION: Quercetin can protect cultured VECs from being injured by high glucose. PMID- 12583179 TI - [Apoptotic morphological changes and TUNEL of human gastric cancer cell SGC-7901 induced by aining]. AB - OBJECTIVE: To investigate the inducing apoptosis effect of Aining on the human gastric cancer cell SGC-7901. METHODS: In vitro cell culture was adopted to explore the inducing apoptosis effect of Aining and cisplation (DDP) through the morphological method and the molecular method. RESULTS: Apoptotic morphological changes were seen after the cells cultured with Aining or DDP; apoptosis rate of 1 g/L Aining group was 46.36%, which had no significant differences with the apoptosis rate 49.12% of the 25 mg/L DDP group; both of the Aining and the DDP groups were significantly different from that of the blank group(P < 0.01). CONCLUSIONS: Both DDP and Aining could induce the apoptosis of human gastric cancer cell and the traditional medicine compound prescription Aining is likely to become a new agent which is used to induce apoptosis of cancer cells. PMID- 12583180 TI - [Hypocholesterolemic effect of total isoflavones from Pueraria lobata in ovariectomized rats]. AB - OBJECTIVE: To observe the effects of total isoflavones from Pueraria lobata (TIP) on lipid metabolism in estrogen deficient rat. METHODS: Female SD rats were bilaterally ovariectomized under anesthesia. TIP was administrated orally for 7 months. Serum lipids were determined at 1.5, 4.0 and 7.0 months, and liver lipids were detected at 7 months. RESULTS: Compared with non-ovariectomized rats, serum TC level and the ration of TC/HDL-C in ovariectomized rats were obviously increased while HLD-C was slightly induced and TG level were changeless. In TIP treated groups, serum TC level and the ratios of TC/HDL-C were markedly lowered than those in OVX group, while HDL-C slightly increased. TG level were much reduced in 1.5 month and changeless in 4.0 and 7.0 month. Slightly increase of cholesterol and markedly induction of triglyceride tissue were observed in liver. CONCLUSION: TIP significantly decreased concentrations of serum TC and liver TG in OVX rats. PMID- 12583181 TI - [Lipid peroxidation of central nerve system in aged guinea pig and antioxidation effect of the method extract from Pegasus laternarius]. AB - OBJECTIVE: To study the lipid peroxidation products level in aged animals central nerve system and the antioxidation effect of the methol extract from Pegasus laternarius. METHODS: The lipid peroxidation product MDA was tested by spectrophotometer. RESULTS: Compared with the 10 month-old guinea pig, the MDA in 32 month-old guinea pig central nerve system obviously elevated, there were some difference in different fields of tested guinea pig brain, the level of MDA in hypothalamus increased biggest (up to 161.7%), cerebral cortex 93.7%, cerebella 84.9%, brain stem 81.2%, spinal cord 90.7%, rest of the cerebrum 58.9%. The method extract from Pegasus laternarius 10, 20, 40 mg/kg and ginseng saponin 20 mg/kg could reduce the level of MDA in tested brain field of aged animals. The method extract from Pegasus laternarius had stronger activity in brain stem, spinal cord and hypothalamus. CONCLUSION: The lipid peroxidation in aged animal may be increased, the method extract from Pegasus laternarius and ginseng saponin have a protective effect on neuron in central nerve system of aged animals from free radical hurt. PMID- 12583182 TI - [Studies on pharmacodynamics of capsule tongluojuanbi]. AB - OBJECTIVE: To study the pharmacological effects of Capsule Tongluojuanbi on antiinflammation and analgesic. METHODS: To apply the rat model of adjuvant arthritis, classical experiment methods of antiinflammation and analgesic. RESULTS: Capsule Tongluojuanbi obviously inhibited primary and secondary plantar swelling of adjuvant arthritis rats, tremendously relieved formaldehyde-induced arthritis in rats, reduced the inflammatory of mouse auricle, reduced the capillary permeability of mouse and diminished twisting reaction of abdominal cavity caused by injection of acetic acid. CONCLUSION: Capsule Tongluojuanbi had obvious effect on antiinflammation and relieving pain. PMID- 12583183 TI - [Internal pharmacokinetical study on genistein chitosan microsphere capsule]. AB - OBJECTIVE: To compare the distribution of genistein chitosan microsphere capsule (M.C) in vivo in rats with that of genistein common capsule (C.C). To research slow-released target microsphere capsule. METHOD: The study on pharmacokinetics of M.C and C.C were analyzed by HPLC and 3P97 software. RESULTS: The concentration of M.C distributed among various organs decreased slower than those of C.C and were able to keep a certain concentration for a long time. Genistein common capsule mainly distributed in livers of rats, while genistein chitosan microsphere capsule distributed mainly in lungs and spleens of rats. CONCLUSION: The chitosan microsphere capsule of genistein has better slow-release and has tendency to distribute in lungs and spleens. PMID- 12583185 TI - [Seedling quality and root rot on Panax notoginseng]. AB - The experiment was carried on the relationship between seedling quality and root rot on Sanqi (Panax notoginseng). The result showed that seedlings from new reclamation land, the root rot disease was fewer and existing rate of seedling was higher than that of seedlings from old gardenplot. Therefore, selecting ill free and robust seedling is one of important measure for the control of root rot on Sanqi cultivation. PMID- 12583184 TI - [A survey of Cornus officinalis varieties]. AB - According to the survey of Cornus officinalis in the main planting areas of Henan, Zhejiang and Shanxi province, the fruit shape can be divided into seven types, cylindricform, ellipticform, long pear-shape, short pear-shape, long cylindricform, short cylindricform and spindlefrom. These fruit types, growth habit and dried pulp productivity can provide scientific basis for breeding cultivation of Cornus officinalis varietis. PMID- 12583186 TI - [Direct identification of Artemisia argyi, Artemisia dubia and Artemisia lavandulaefolia by FTIR]. AB - OBJECTIVE: To directly and accurately identify Artemisia argyi Levl. et Vant., Artemisia dubia Wall. and Artemisia lavandulaefolia DC. METHODS: Fourier tansform infrared (FTIR) spectrum method was used. RESULT: There were obvious differences among the FTIR spectrums of above-mentioned plants. CONCLUSION: Artemisia argyi Levl. et Vant., Artemisia dubia Wall. and Artemisia lavandulaefolia DC. were directly, fast and accurately identified by FTIR. PMID- 12583187 TI - [Identification on Chinese materia medica Radix Rehmanniae preparata by X-ray diffraction Fourier pattern method]. AB - OBJECTIVE: To develop a new identification and analysis method of Chinese medicinal materia Radix Rehmanniae Preparata. METHODS: Powder X-ray Diffraction Fourier Pattern. RESULTS: Experiments and analysis were carried out on five samples. The standard X-ray diffraction Fourier Pattern and characteristic diffraction peaks of Radix Rehmanniae Preparata were obtained. CONCLUSION: This method can be used for identification on Chinese medicinal materia Radix Rehmanniae Preparata. PMID- 12583188 TI - [Study on effect of air-adjustment storing ginseng]. AB - The research of air-adjustment storing ginseng shows that when the concentration of CO2 is above 50% and the concentration of O2 is under 5%, all adults will die in four days; all eggs and larvas will die in fifteen days. During the year of using this method air-adjustment, no vermins and mould have been found. PMID- 12583189 TI - [Study of chemical constituents of the essential oil form Pericarpium Zanthoxyli by GC-MS]. AB - The chemical components of the essential oil from Pericarpium Zanthoxyli collected from Jinan city of Shandong province and Hengshui city of Henan province were analyzed by GC-MS. 36 components from Pericarpium Zanthoxyli in Jinan city of Shandong province that composed of about 87% of the total essential oil were separated and identified. 20 components from Pericarpium Zanthoxyli in Hengshui city of Henan province that composed of about 74% of total essential oil were separated and identified. PMID- 12583190 TI - [Study on the biological assay of Herba Leonuri(I)--Establishment of standard uterus models and optimizing uterus environment conditions]. AB - OBJECTIVE: To study the contracting uterus effect of Herba Leonuri, establish the standard uterus models, and optimize the environment conditions which could influence Herba Leonuri actions on uterus. METHODS: By comparing the contracting uterus effect of Herba Leonuri in mice, rats and rabbits, the standard uterus models was established. The optimum extra conditions of uterus were selected by orthogonal design methods. RESULTS: The isolated rat uterus pre-oestrus was selected for the standard uterus, the optimum uterus environment conditions as follows: the pre-burden of uterus was 1.0 g, the temperature of uterus nutrition solution is 32 degrees C, the assay of CaCl2 and NaHCO3 per 1000 ml nutrition solution was respectively 0.06 g and 0.25 g. CONCLUSION: Under the selected conditions, the contracting uterus effect of Herba Leonuri were more obvious and the errors of bioassay were less smaller. PMID- 12583191 TI - [Effect of sinamine on withdrawal symptom and neurotransmitter of morphine dependent rats]. AB - OBJECTIVE: To study on the detoxification effect of sinamine in morphine dependent rats. METHODS: Morphine-dependent rats were induced by injecting morphine on dosage increasing by degrees, then treated with medication. The withdrawal symptoms, body weight and NE, DA, 5-HT in the brain were tested. RESULTS: Sinamine could alleviate withdrawal symptom, reablement body weight, inhibit neurotransmitter in the brain. CONCLUSION: Sinamine have effects on morphine-dependent rats which may relate to modulating neurotransmitter. PMID- 12583192 TI - [Morphological changes and inhibiting effect on human gastric cancer cell SGC 7901 caused by aining]. AB - OBJECTIVE: To investigate the inhibiting effect of Aining on the human gastric cancer cells. METHODS: Morphological and MTT methods were adopted to explore the inhibiting effect of Aining and cisplatin (DDP) on the proliferation of SGC-7901 cancer cell. RESULTS: Apoptotic morphological changes were seen after the cells cultured with Aining and DDP; inhibiting rate of 1 g/L Aining group was 51%, which had no significant differences with the inhibiting rate 53% of the 25 mg/L DDP group. But both the Aining and the DDP groups were significantly different from the blank group (P < 0.01). CONCLUSION: Not only DDP but also Aining could inhibit the proliferation activity of the human gastric cancer cells, and the traditional Chinese medicine compound prescription Aining is very likely to become a new medicine which is utilized to inhibit cancer. PMID- 12583194 TI - [Effects of Angelica polysaccharide on blood coagulation and platelet aggregation]. AB - OBJECTIVE: To investigate the effects of angelica polysaccharide (AP) on blood coagulation and platelet aggregation. METHODS: Infrared turbidimetric method was used to estimate platelet aggregation, active partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT); bleeding time (BT) was measured by cutting the mouse's tail and coagulate time (CT) was measured by dropping the blood on carry sheet glass. RESULTS: AP prominently enhanced the platelet aggregation at 5 min, while showed less effects on the maximum platelet aggregation. It also markedly prolonged CT but shortened BT. AP significantly prolonged TT and APTT in dosage of 3 mg/kg and 10 mg/kg, while showed no obvious effect on PT. CONCLUSION: These results suggest that AP has potent anticoagulant and haemostasis effects. The haemostasis effect is related to promoting platelet aggregation. PMID- 12583193 TI - [Effect of total glycoside from Clinopodium polycephalum on uterine bleeding quantity in drug abortion model rats]. AB - OBJECTIVE: To study the effect of total glycoside from Clinopodium polycephalum (Vaniot) C. Y. Wu et Hsuan on uterine bleeding quantity in drug abortion model rats. METHOD: Mifepristone and misoprostol were given to early-pregnancy rats orally. The change of uterine bleeding quantity was observed in uncompletive abortion model rats. RESULT: TGCP (20 mg/kg) could markedly reduce uterine bleeding quantity in drug abortion model rats. CONCLUSION: TGCP has the hemostatic effect on uterine bleeding in drug abortion model rats. PMID- 12583196 TI - [Study on enrichment process of Cornus officinalis glycosides with macroporous resin and active carbon]. AB - OBJECTIVE: To study the technological parameters of the enrichment purification of Cornus officinalis glycosides (COG) with macroporous resin and active carbon. METHODS: COG was isolated and purified by active carbon and macroporous resin HPD100. The enrichment of COG was evaluated by HPLC method. RESULTS: The optimum process was that 10 ml extract of Fructus Corni was absorbed with columns of macroporous resin and active carbon(R25 mm x H300 mm) and the columns were washed with 600 ml distilled water, then the morroniside and loganin was eluted from the macroporous resin and active carbon with ethanol. CONCLUSION: The elutive rate of morroniside and logann was about 85% by the macroporous resin, 96% by the active carbon. So this method enriching COG from Fructus Corni is practical. PMID- 12583195 TI - [The research of 10-hydroxy-2-decenoic acid on experiment hyperlipoidemic rat]. AB - OBJECTIVE: To study the pharmacological effect of 10-hydroxy-2-decenoic acid(10 HDA) in experiment hyperlipoidemic rat. METHODS: Preventive and therapeutic effects of 10-HDA were tested on hyperlioidemic rat model induced by high fat food. RESULTS: 10-HDA could reduce the content of TC, TG and beta-lioprotein, raise the content of HDL, which showed 10-HDA had preventive and therapeutic effects on hyperlipoidemic rat. CONCLUSION: 10-HDA was functional factor of preventive and therapeutic effects of royal jelly on hyperlipoidemia. PMID- 12583197 TI - [Development of a portable electrocardiogram(ECG) and respiration monitor]. AB - This paper is to research into a kind of portable automatic ambulatory system, which is used to monitor electrocardiograph (ECG) and respiration signals. And in the interest of meeting the requirements of continuously monitoring the patients, comprehensive consideration is given to the overall structure of the system (including the display mode and the management of power supply). With the characteristics of higher sensitivity and preferable anti-interference, this system realizes minialurization and low power dissipation. PMID- 12583198 TI - [The application of adaptive line enhancement filter in extracting cutaneous electrogastrogram]. AB - Based on the principle of adaptive filtering, The higher signal-to-noise ratio of electrogastrogram data can be extracted from abdomen skin with the adaptive line enhancement filter. The design method of feedback factor and filtering coefficient is introduced in this paper. Experimental and practical results show the method is effective. PMID- 12583199 TI - [Signal and data acquisition based on waveform graph]. AB - It is well known that signal acquisition is the first step in signal processing. A simple and convenient method of acquiring biologic data is described here, which needs no special acquisition equipment, and is practical and makes the data acquisition more credible. PMID- 12583200 TI - [Research into estimating lumen center of dark region in colon image via cubic B spline wavelet]. AB - In this paper, a new method is presented to detect the edge of a colonscope image using cublic B-spline wavelet. This method can make up for the shortcoming of traditional method, resulting from blue dark area in colonscope image through estimating the circle centers. These estimated circle centers which have arc shaped edge lines are weighted to obtained the lumen centers. This center is used to navigate colon endoscope in colonscopic operation. PMID- 12583201 TI - [High-performance dynamic ECG review & analysis system]. AB - This paper describes the design of a high-performance dynamic ECG Review & Analysis system used for not only clinical diagnosis but also medical research. A new ECG pattern classifier with Variable Area Template is applied to improve the quantity of ECG analysis results, while the optimized software design ensures the other performances. The results show that this system has higher performance than the old ones and is valuable for medical research. PMID- 12583202 TI - [Feature analysis for the envelope of liver's M-mode ultrasound images]. AB - The liver moves in response to impulses arising from the main blood vessels. The hardness of the liver may be reflected by this kind of movement. The M-mode ultrasound technique is used in this paper to detect the information of livers' motion. The motion envelope of the liver excited by the abdominal aorta is extracted using the correlation technique. Several feature parameters, such as the amplitude, the standard deviation and Fourier harmonics, are extracted from this envelope. The values of parameters were calculated from 19 normal livers and 11 patients' livers, from which the criterion was obtained to decide whether the liver was normal. PMID- 12583203 TI - [The design of ultrasound subsystem in PACS/HIS environment]. AB - Playing an important role in PACS, ultrasound subsystem together with CT and X radiography subsystems works as the image Provider and image processing center. In this paper, we propose a novel scheme on how to construct ultrasound subsystem and explain it with detailed work flow and software frame. We also give a discussion on how to expand such a subsystem. PMID- 12583204 TI - [The development of computerized cardiac programmable stimulator]. AB - This paper introduces a fully computerized cardiac programmable stimulator. With the visual interface of computer, the configuration and control of stimulation will be realized conveniently. Meanwhile, the current status and the waves during stimulation will be displayed in window real-time, which improved the visualization and availability in medical application. It has been proved that this system is simpler in operation and more functional than traditional cardiac programmable stimulators, and therefore it is valuable in medical applications. PMID- 12583205 TI - [The design of detection circuit for DNA biosensor]. AB - The paper describes the design of a quartz crystal sensor for the detection of DNA hybridization reaction. With the ssDNA as probe on the electrode surface, we can detect the frequency change and show it on the computer display. The design is capable of real time monitoring for frequency change of DNA hybridization reaction. PMID- 12583206 TI - [The development of a residual voltage tester]. AB - The paper introduces the key points of the residual voetage tester's designing and its circuit frame diagram. PMID- 12583207 TI - [The development of dental silver amalgam glass ionomer cement]. AB - To eliminate the harm of mercury, we, by means of special technology, have developed a new type of dental filling material, comprising of glass ionomer and 100% ultra-fine ball-shaped silver amalgam powder. Clinical applications show it has very good wear-resistant, pressure-resistant and adhesive properties Moreover, it causes no irritation to the dental pulp and is a good kind of dental filling material in clinical applications. PMID- 12583208 TI - [The study of the peritoneal dialyzer with multipriority and its auxiliary spring brush]. AB - This paper introduces the design, principle and clinical applications of the peritoneal dialyzer with multipriority and its auxiliary spring-brush. PMID- 12583209 TI - [Developmental trend & network exploitation of central monitoring system]. AB - This paper briefly describes developmental trend and network exploiting situation of central monitoring system, analyzes preliminarily its subsequent developmental dynamic. PMID- 12583210 TI - [The application of programmer & tester in the repairing of intelligent medical instrumentation]. AB - This paper introduces the functions and the features of programmer & tester. In repairing intelligent medical instruments, which contain digital circuit or micro processor interface circuit, it the programmer & tester is properly used, it will help to prompt obviously the repair level and repair efficiency. PMID- 12583211 TI - [The principles and developments of the hemodialysis system]. AB - This paper describes the machinery and basic theory of a dialysis machine, and presents some of the latest developments. PMID- 12583212 TI - [Hemodialysis units and the optimal dialysis process]. AB - This paper describes the therapy of kidney failure, the principles of a hemodialysis unit ant its development, and also introduces the optimization of dialysis process by real-time monitoring of Hct and SaO2. PMID- 12583213 TI - [Analysis and detection of abnormal ECG signal: ventricular tachycardia and fibrillation]. AB - This paper, utilizing signal power spectrum and L-Z complexity measure, analyzes the abnormal ECG: ventricular tachycardia (VT) and ventricular fibrillation (VF). And on the base of linear and nonlinear dynamics, the paper forwards a new definition of information complexity rate, deduces some relative properties and applies it to complexity analysis in irregular ECG. By means of biological experiments and computer simulations, the reasonability of information complexity and complexity rate is confirmed. At last, objective analysis and explanations of the mechanism of VT and VF are reported. The results indicate: with the help of power spectrum, complexity measure and complexity rate, the recognition of VT and VF can attain 100%. PMID- 12583214 TI - [Rigid registration and fusion between SPECT and MRI/CT based on external fiducial alignment]. AB - A convenient and effective method of rigid registration and fusion between SPECT and MRI/CT which is based on external fiducial alignment system is designed. After thresholding the image, many image processing arithmetics such as erosion, dilation are used to ensure the corresponding points which will be used to registrate. Then these points are used to minimize the cost function. In this way, an optimal rigid transform is gotten to registrate the images accurately and fuse them well. Error in the registration and fusion is reduced furthest by well designing the N-shape bracket which is used as the external fiducial alignment system and also by limiting the relative displacement between the bracket and the patient. Experimentation shows that this method can realize the registration and fusion of SPECT and MRI/CT effectively. PMID- 12583215 TI - [The development of cerebral circulation analyzer]. AB - In this paper a new cerebral circulation analyzer is introduced, inducing the main structure, the operating principle, the software program and its clinical applications. We can get the cerebrovascular hemodynamics indexes such as resistance, compensatorg blood flow etc., from the blood velocity, pressure waveform, and arterial diameter detected in carotid and vertebral arteries. PMID- 12583216 TI - [Picking up and analysis of the surface myoelectric signals of respiratory muscule]. AB - In this paper, we introduce the technique used to obtain the surface diaphragmatic EMG and monitor respiratory activity. The signals are picked up from the ECG electrodes. By using an ECG masking system based on a digital processor, the dominant effect of the ECG (that is R-wave, P-wave and T-wave) was removed. Initial clinical measurements indicate this EMG method is more direct and effective than others for monitoring respiratory activity. It is hoped that this method can be used to monitor the development of respiratory function. PMID- 12583217 TI - [Research on automatic diagnosis system of electrocardiogram]. AB - The duration of QRS wave, P wave and T wave of ECG can be computerized and auto detected with digital signal process and wave recognition technology. It can diagnose 15 kinds of abnormal ECG such as ventricular presystole automatically and print out corresponding diagnosis reports. In order to verify the system's stability and creditability, we used American MIT-BIH database to test our algorithms and got a good result. PMID- 12583218 TI - [Realization of voice coil motor control with H8/338 micro-controller unit in left ventricular assist pump]. AB - In this paper, we present a PID algorithm with velocity project of voice coil motor controlled on the base of H8/338MCU, and we find a driving part, which it is smaller, lighter and more reliable, for the left ventricular assist device. By means of the simulated experiment on the system, the result is satisfying. So we can say that it is a good help for the realization of LVAD. PMID- 12583219 TI - [Noninvasive measurement of gastric emptying rates and gastric motility]. AB - A non-invasive measuring system is developed for the assessment of gastric evacuation and motility by means of epigastric impedance measurement. FIR digital filters realized on personal computer were used for signal filtering and cancellation of respiratory interference. Electrodes of concentric type placed anterior and posterior were proved to be more effective for epigastric impedance measurement. PMID- 12583220 TI - [A implementation for digital scan converter(DSC) by using FPGA]. AB - A design of a Digital Scan Converter(DSC) in B scanner and its implementation by using FPGA are presented in this paper, and some applied results are presented too here. PMID- 12583221 TI - [The development of body posturography device using inclinometer technique]. AB - A kind of body posturography device using inclinometer technique is described in this paper. Comparing with a device using gravimeter technique it shows following advantages: 1) As the signal of body sway angle is obtained by the incline sensor, it is easy to test subject's balance function on the multifarious body supporters; 2) When the platform of the global bottom is used, the subject may come into contact with ground at one point, so as to weaken significantly subject's somatosensory of moving center of gravity. PMID- 12583222 TI - [A simple X-ray TV picture freezer]. AB - A simple X-ray TV picture freezer, with just 5 chips of LSIC, is discussed in this paper. Its VRAM block is made up of cheap and relatively lower speed large scale SRAM. The memory block works in alternate mode and can meet the sample speed up to 10-12 Msps and the pixel matrix of 512 x 512. PMID- 12583223 TI - [Probe into the virtual medical instruments]. AB - With the use of 3DSMAX and AUTO CAD 2000, we have practiced the dynamic simulation of the virtual medical instrument, which is very useful in design. The engineer and doctor can watch a product and change it before making so as to save the time of design and to improve the quality. PMID- 12583224 TI - [Multi-slice CT scanners]. AB - Some basic specifications of four commercial multi-slice CT scanners are compared with each other. The new technologies under the development of multi-slice CT are described and the image quality performance of multi-slice is discussed in this paper. PMID- 12583225 TI - [The late achievements of volume CT]. AB - This paper mainly introduces the late achievements of theories and methods for this technology. And we look into the future of them at the end of this paper. The main contents include direct volume CT and its fast algorithms, volume imaging for non-complete data, direct volume CT based on digital radiography and reverse geometry X-ray. PMID- 12583226 TI - [Digital radiography detector]. AB - This describes the detectors for digital radiography and the current state of flat panel detectors(FPD) especially. PMID- 12583227 TI - [The manufacture of the pressure transducer for medical Solos Insufflator]. AB - A 0-10 Kpa pressure transducer and its circuit are presented in the paper, which can be used as a substitute for the integrated pressure transducer in medical Solos Insufflator and can be used too in other devices and equipments. PMID- 12583228 TI - [A simple coupled instrument for hyperthermia]. AB - In this paper, we present a temperature-controlled watercycling coupled instrument, which changes from the air-couple of the applicator to water-cycling couple when superficial heating is given. This instrument in combination with the characters of the present microwave device, has brought about the changes of coupling media without changing the structure of the applicator. PMID- 12583229 TI - [Disposition of water-supply pipeline and cleaning & disinfection of reverse osmotic membranes in the department of hemodialysis]. AB - In this article we present some prescriptions & methods of cleaning & disinfection for osmotic membranes and also exhibit the benefits and limitations of the two water-supply pipelines. PMID- 12583230 TI - [A method of determine the inspection period of measuring instruments in a hospital]. AB - In this paper we apply the principles of statistics to analyze a measuring instruments of diagnosis and treatment, and then make overall evaluations and quantifications so as to determine the inspection period of a measuring instrument. PMID- 12583231 TI - [Study on application of arbuscular-mycorrhizas in growing seedings of Aloe vera]. AB - Tissue culture seedlings of Aloe vera L. inoculated with 7 AMF(arbuscular mycorrhiza fungi) in a greenhouse in Guangzhou showed that the percentage of infection was 99.67%-100%, the index of infected was 73.3%-86.67%. After being inoculated 13 months, the seedling high raised 19.88%-51.91%, the leaves length raised 13.13%-150.96%. After being inoculated 15 months, the leaves juice of Aloe vera raised 60.87%-233.8% and the dried of leaves juice raised 217%-724%. PMID- 12583233 TI - [Study on dry matter accumulations of Houttuynia cordata on heavy loam]. AB - In this paper the characteristics of dry matter accumulation of Houttuynia cordata cultivated on three kinds of heavy loams were studied. The results showed that the accumulated curves of whole plant dry matter were somewhat different. During testing period, the accumulated curves of dry matter of Houttuynia cordata grown on clay silt loam and silt sand loam presented logistic ones, but those grown on sandy silt loam were still in line growth period. It was concluded that the harvest period of Houttuynia cordata on the sandy silt loam could be properly put off while only considered the yielding. The experiment on heavy loam also showed that the sandy field should be the better selection than others, but organic fertilizer should be enhanced as base fertilizer, with proper nitrogen as top application. This paper also discussed the dry matter accumulations of different organs of Houttuynia cordata. PMID- 12583232 TI - [Study on seedling inducement from seed of Anoectochilus roxburghii]. AB - This experiment showed that the best medium for protocorm inducement from seed of Anoectochilus roxburghii was Knudson C, the suitable medium for protocorm growing to young plant was Knudson C + 6-BA0.5 mg/L + 20% extract of potato. The reproductive rate of seed can achieve 51%. PMID- 12583234 TI - [Bionomics and breeding of ant-lion]. AB - Ant-lion is a Chinese crude drug. The natural factors, such as temperature, humidity, illumination, etc, have an obvious influence upon its growth and movement. The temperature fit to its growth ranges from 25-30 degrees C and lighting 14-16 h/d. If giving enough food, it can reproduce 3 generations per year. PMID- 12583235 TI - [Study on morphology of the pollen of 4 species from Geranium in Heilongjiang Province]. AB - Morphology of the pollen of 4 species from Geranium in Heilongjiang province which are G. wilfordii Maxim., G. sibiricum L., G. platyanthum Duthie. and G. dahuricum DC. were observed with optical microscope and scanning electron microscope. The results showed pollen grains of the four species were nearly spherical, with short clavate-shaped, net-shaped and cave-shaped veins on the outer walls and three short-furrow germ pores. Both the clava size and number of clava per unit surface area were different. This paper provided evidences for palynology classification of Geranium. PMID- 12583236 TI - [Constituents analysis on volatile oil of Pogostemon cablin from different collection time cultivated in Hainan]. AB - The constituents of the volatile oil of Pogostemon cablin collected from Wanning city, Hainan province and the influence owing to different collection time were undertaken by GC/MS combination technology. The results showed that main compounds was patchouli alcohol in stem oil and leaf oil, its contents were respectively 36.06% and 37.74%; pogostone was poor, its contents in stem oil and leaf oil were respectively 17.08% and 0.85%; the contents of ten compounds were over 1%, which was respectively beta-patchoulene, beta-elemene, transcaryophyllene, delta-guaiene, seychellene, alpha-patchoulene, aciphyllene, alpha-guaiene, patchouli alcohol and pogostone; the contents of volatile oil from June to August were respectively 0.8%, 0.7% and 0.6%; patchouli alcohol, higher in July and June(42.62% and 40.84%), lower in August(31.40%). PMID- 12583237 TI - [Determination of ampelopsin in the different parts of Ampelopsis grossedentata in different seasons by RP-HPLC]. AB - OBJECTIVE: To determine ampelopsin in the root, stem and leaf of Ampelopsis grossedentata in different seasons by RP-HPLC. METHODS: The determination was carried out with ODS column (5 microns, 250 mm x 46 mm), using methanol-water phosphoric acid(25:75:0.1) as the mobile phase at a flow rate of 1.0 ml/min and detected at the wavelength 289 nm. RESULTS: Ampelopsin content was in the order of leaf > stem > root. Ampelopsin content in Ampelopsis grossedentata collected in April and May was the highest. PMID- 12583238 TI - [Isolation, purification and analysis of polysaccharides from Adenophora potaninii Korsh]. AB - Polysaccharides, AP-1 and AP-3, were obtained through extraction of the roots of Adenophora potaninii Korsh by hot water, followed by the isolation and purification by DEAE-cellulose and Sephadex G-200 column chromatography. It was proved to be homogeneous by gel filtration chromatography and their average MW was estimated to be 8.3 x 10(4) and 6.3 x 10(4) respectively. Analysis of sugar composition showed that AP-1 is D-glucan, while AP-3 is composed of glucose, rhamnose, arabinose and glucronic acid in the molar ratio of 5:1:1:3. PMID- 12583239 TI - [Protective effect of tea pigments on mice skin photoaging induced by UV irradiation]. AB - OBJECTIVE: To study the protective effect of tea pigments on mice skin photoaging induced by UV irradiation. METHOD: Total antioxidative ability and content of lipid peroxidation products malondialdehyde in the skin of mice were examined with colorimetry. RESULT: By ig or smearing, tea pigments could significantly improve total antioxidative ability and decline content of malondialdehyde in UV irradiation-induced skin photoaging mice. CONCLUSION: The results suggested that tea pigments could prevent mice skin from free radicals injury resulting from UV irradiation and moderate skin photoaging. PMID- 12583240 TI - [Effect of tanshinone IIA on CCl4-induced liver fibrosis in rats]. AB - OBJECTIVE: To study the effect of tanshinone IIA on liver fibrosis in rats induced by CCl4. METHODS: Model of liver fibrosis in rats were induced by subcutaneous injection of CCl4 and malotilate was as positive control. The histological changes were observed with light microscope. The levels of ALT, AST, NO, HA, LN in serum and Hyp, MDA in liver tissue were determined to reveal the event liver damage. RESULT: Tanshinone IIA could significantly reduce the elevate levels of ALT, AST, NO, HA, LN in serum and Hyp, MDA in liver tissue. Pathological examination suggested tanshinone IIA has therapecuical effect on experimental liver fibrosis. CONCLUSION: Tanshinone IIA may have therapeutical effect on CCL4-induced liver fibrosis in rats. PMID- 12583241 TI - [Effect of the stem and leaf of Tripterygium wilfordii on immune function]. AB - OBJECTIVE: To study the effect of water extracts from the stem and leaf of Tripterygium wilfordii Hook. f. on Hook. f. immune function. METHODS: The effect of the stem and leaf of Trpterygium wilfordii on the clearance of charcoal particles, the index of thymus gland and spleen, the level of serum hemolysin and the delayed hypersensitivity were observed in mice by ig. RESULTS: The stem and leaf of Tripterygium wilfordi could decrease the clearance of charcoal particles, the index of thymus gland and spleen, the level of serum hemolysin and inhibit the delayed hypersensitivity in mice. CONCLUSION: The stem and leaf of Tripterygium wilfordii could inhibit nonspecific, humeral and cellular immunity. PMID- 12583242 TI - [Effects of Pedicularis decora extract on anti-oxidation injury in mice]. AB - OBJECTIVE: To study the protection effects of Pedicalaris decorm extract on oxidation lesion in mice and the effective components of Pedicalaris decorm. METHODS: The total extract, BuOH soluble components and water soluble components of Pedicalaris decorm were administered (ig) 15 g.kg-1.d-1 to mice, and the oxidation injury model in mice was induced by i.p. alloxan 180 mg.kg-1. SOD, GSH Px and LPO in red blood cell and liver, ALT, AST, AKP and r-globulin in serum, LPO and MAO.B in brain, liver index and spleen index were measured. RESULTS: All the three Pedicalaris decorm extracts can obviously lower abnormal contents of SOD, GSH-Px, LPO, ALT, AST, AKP, MAO.B and r-globulin induced by alloxa, and obviously improve abnormal decrease of liver index and spleen index. CONCLUSION: Pedicalaris decorm extract can inhibit oxidation lesion in mice. The effects of total extract and BuOH soluble components were better than water soluble components. PMID- 12583243 TI - [Study on reducing blood sugar of polysaccharide from Cyclocarya paliurus (Batal.) Iljinsk]. AB - OBJECTIVE: To try to find Cyclocarya paliurus (Batal.) Iljinsk contains the polysaccharide complex with reducing blood-sugar activity or not. METHOD: The Polysaccharide was extracted by ultrasonic wave. The diabetes model was induced by alloxan to verify the activity of the polysaccharide. RESULTS: The content of the polgsccharide is 9.6% in Cyclocarya paliurus. The blood-sugar level of the polysaccharide group is lower by 41.3% than that of the diabetes model group. After ig glucose, the blood-sugar level of the pollysaccharide group was obviously lower than that of the diabetes model group. CONCLUSION: Cyclocarya paliurus (Batal.) Iljinsk is rich in polysaccharide complex, which have obvious reducing blood-sugar activity and can improve the capability of glucose tolerance in diabetes mice. PMID- 12583244 TI - [Herbalogical study on Plantago asiatica L]. AB - According to textual study of herbal drugs and modern scientific research of chemical constituents and pharmacological actions of Plantago asiatica L., the record in the ancient literatures was found to be basically identical to that obtained by modern scientific research. Plantago asiatica from Plantaginaceae family is regarded as the orthodox medicinal herbs through the ages. PMID- 12583245 TI - [Superfine comminution technology and its application in the processing of Chinese traditional medicine]. AB - The definition and mechanism of superfine comminution is introduced and the characteristics of equipments in common use for superfine comminution were outlined. The advantages of Chinese traditional medicine processed by superfine comminution are analyzed. The stress was laid on the present state and their advantages and disadvantages of the equipments used in the processing of Chinese traditional medicine. Vibration mill was pointed out to be the optimal equipment for the superfine comminution of Chinese traditional medicine. PMID- 12583246 TI - [Clinical study of treatment on chronic uric acid nephropathy by integrating Western and traditional Chinese medicine]. AB - OBJECTIVE: To investigate the ameliorative effect on chronic uric acid nephropathy (CUAN) by integrating western and traditional Chinese medicine (IWTCM). METHODS: The 136 CUAN patients were divided into two groups at random, the therapy group of 86 patients were treated by Chinese medicine and allopurinol, and the control group of 50 patients were treated only by allopurinol. The curative effect and the related index such as blood uric acid, renal function, urinary protein, microproteins, blood lipid and hyperviscosity were determined before and after being treated. RESULTS: After three months treatment, the total effective rate in the therapy group (90.7%) was significantly higher (P < 0.01) than that of the control group (56%). The therapy group is also superior to the control group in improving renal function, lipid metabolism and hyperviscosity, decreasing blood uric acid, urinary protein, microproteins in evidence (P < 0.05, P < 0.01). CONCLUSION: IWTCM can obviously improve the ameliorative effect on chronic uric acid nephropathy. PMID- 12583247 TI - [An computerized fundus atlas system]. AB - This paper presented an computerized fundus atlas system for WINDOWS. The classical normal and abnormal fundus photoes were digitalized and saved in real color image format. The graphics technology was used to label the area and the feature of the abnormal section with WINDOWS atom functions. When these functions run, the graphics and the texts, which label the area of the selected section and describe their features, were displayed in a carton mode background funds image. PMID- 12583248 TI - [The disposing techniques of evoked potentials]. AB - This paper is to bring forward the new disposing techniques of evoked potentials which include four aspect techniques of the averaging, the recording, digital sampling and filters about the averaging, evoked potential amplitude, evoked potential latency, evoked potential recording, and evoked potential generations. The technique of the averaging including signal filtering and a periodic averaging, can enhance EP dedection. The commercial EP machines also plot changes in latency between serial EP studies in order to detect trends in peak latency. The modern digital EP recording device consists of sensory stimator, recording amplifiers with analog filters, an analog-to-digital converter, a digital signal averager, and a display and storage system. A sample-and-hold function is one of the recent developments which used EP collectors that provide simultaneous recording with multiple channels employing different time and voltage scales and sampling rates. The EP data may be further processed following A-D conversion by digital filters. PMID- 12583249 TI - [The development of an instrument for testing body water equillibrium in hemodialysis patients]. AB - In order to obtain a real-time and correct test for body water equilibrium in hemodialysis patients, an instrument based on bioimpedance measurement has been developed. The hardware and software design of the instrument was introduced in this paper. The instrument is of safty and good duplicity. Preliminary clinical experiments shows that it can reflect the state of body water with relative high sensitivity. PMID- 12583250 TI - [Development of the portable blood glucose meter]. AB - This article introduces the research work on blood glucose assay instrument. It uses the method that immobilizes enzyme on nylon66 film and detects resultant color by photo-electric sensor The detecting range is 0-30 mml/L and the detecting time is 50 s. PMID- 12583251 TI - [A study of temperature rising during the excimer laser keratectomy]. AB - Here, we try to study the regularity of the temperature changing in radiated materials during excimer laser pho-torefractive keratectomy, and more attention should be paid to it during the excimer laser keratec[symbol: see text]tomy. PMID- 12583252 TI - [The sparkle noise abatement in RF excitation signal of LMW-400 MRI system]. AB - We found a sparkle noise in the Radio-Frequency excitation signal of LMW-400 MRI system manufactured by Guangdong Weida Medical Apparatus (Group) Corporation. This sparkle noise appeared just following the RF pulse and had a maximum level of about 40% of maximum RF pulse level. Furthermore, since this sparkle noise existed during the read-out gradient period, it must be eliminated in order to reduce image artifacts. Tests on the Modulation Board showed that this noise was inherent due to abrupt rising edges of two control. signals. Simple integrating circuits had been applied to those two signals and the sparkle noise could be eliminated successfully by the modification. PMID- 12583253 TI - [The research on processing method of infrared thermography for diabetes]. AB - The article shows the author's exploratory opinion about the processing method of infrared thermography for the diagnosis of diabetes. PMID- 12583254 TI - [The study and manufacture of multifunctional brain microsurgery operation dilator]. AB - The dilator consists of fixative clasp-handle, fixative platform, support arm, spatula and handle. The dilator may be fixed on the edge of bone window easily in the brain operation. The functions of the dilator include localization, dilation, expose, washing, lighting and suction. The spatula of the dilator may be prepared according to the different depth and width of the operation field. PMID- 12583255 TI - [The newest progress of the breath diagnosis system]. AB - This paper presents the recent international progress of the breath diagnosis systems. It discussess the basic principles and method of the breath diagnosis. Especially, the paper introduces the newest progress and the developing trends in diagnosis of the diabetes and gastrointestinal diseases etc. Finally, it introduces the progress of the medical diagnosis expert system and its application in the breath diagnosis based on the electronic nose. PMID- 12583256 TI - [Digital technology of X-ray radiography]. AB - This paper introduces to you some methods to radlige the digital radiogzaphy, and there respectiae princiales and featuus. PMID- 12583257 TI - [Metallic stents for malignant esophageal neoplasms stenosis in advanced stage]. AB - Maligant esophageal stenosis affects the qulity of the patients' life severely. Self-Expanding Metallic Stent (SEMS) can relieve the patient from dysphagia. In this article, we summarized the application and clinical complication of SEMS in maligant esophageal stenosis. PMID- 12583258 TI - [Techniques of quantitative measurement of intracellular free Ca2+ concentration]. AB - For individual cell, calcium is a signal of life and death and many functions are mediated by it. The development of new fluorescence dye and imaging technique have greatly promoted the research in the field of cellular calcium signal transduction path. PMID- 12583259 TI - [Ion single channel signal restoration and parameters' estimation based on the hidden Markov models]. AB - The single ion channel signal is stochastic ionic current on the order of 1 pA recorded by patch clamp. Because the weakness of the signal, the background noise always dominates in the recordings, the threshold detector traditionally used in patch clamp to denoise and restore the channel signal can't work satisfactorily. This problem was analyzed mathematically, and a signal restoring and parameters estimating scheme called HMM algorithm was studied. The algorithm has been validated by simulation and the results suggest it performs effectively in the situation of low signal to noise ratio where the threshold detector fails completely. PMID- 12583260 TI - [Non-linear compression for noises in non-contact vital signs detecting system]. AB - A method for compressing the noises overlapping in the breath and heart signals of human beings, detected by the non-contact vital signs detecting system during the display of the waveforms in time domain, is discussed in this paper in detail. And what's more, the problem that the noise level is changed along with the increase of the gain of AD data-acquisition card and the display--gain of the software is solved by researching into the threshold of noise-level contraction in two types of circumstance in the lab, and the result of the breath and heart signal in the condition of lower noise is realized as well. PMID- 12583261 TI - [The objective detection and description of the types of pulse based on the Chinese traditional medical science]. AB - Through the study of the classical works of "The Pulse Studies of Binhu", the possibility and methods of the realization of the objective detection and description of all types of pulse based on the Chinese Traditional Medical Science(CTMS) have been proved, and a design scheme of a complete atlas of the types of pulse based on the CTMS is presented in this paper. PMID- 12583262 TI - [The design of near infrared mammography computer-aided diagnosis system]. AB - Near infrared mammography is a new method for breast imaging for popular examination. Its merits are no radiation, without the necessaries of darkroom, etc. Based on the digital image processing and analysis system, the doctor can get informations more conveniently and with higher diagnosis reliability. In this paper, the near infrared image acquisition, processing and analysis system and its implementation are presented. PMID- 12583263 TI - [An experimental study of the computer-controlled equipment for delivering volatile anesthetic agent]. AB - Our newly-designed computer-controlled equipment for delivering volatile anesthetic agent uses the subminiature singlechip processor as the central controlling unit. The variables, such as anesthesia method, anesthetic agent, the volume of respiratory loop, age of patient, sex, height, weight, environment temperature and the grade of ASA are all input from the keyboard. The anesthetic dosage, calculated by the singlechip processor, is converted into the signals controlling the pump to accurately deliver anesthetic agent into respiratory loop. We have designed an electrocircuit for the equipment to detect the status of the pump's operation, so we can assure of the safety and the stability of the equipment. The output precision of the equipment, with a good anti-jamming capability, is 1-2% for high flow anesthesia and 1-5% for closed-circuit anesthesia and its self-detecting working is reliable. PMID- 12583264 TI - [The development of SPA bone densitometry for heel bone]. AB - The design principles of a bone densitometer for heel bone based on SPA are introduced here, which include the principles of mechanical system and control system. The main technical index and advantages of the device have also been discussed in the paper. PMID- 12583265 TI - [Application of differential wavelet in detecting QRS complexes of ECG signal]. AB - An algorithm based on wavelet transforms (WT'S) has been developed for detecting QRS complexes of ECG signal. In this paper, we use Mallat algorithm to calculate dyadic Marr wavelet transforms, analyse the relationship between singularity of the signal (the peak of R wave) and local maximal value of WT's from the view of equivalent wavelet filter, discuss the performance distinction between quadratic differential wavelet and single differential wavelet for singularity detection. Some other tactics are also applied to improve the anti-interference performance of the algorithm and to increase the correct detection rate of QRS complexes. Verified by the MIT-BIH Arrhythmia Database, the correct detection rate of QRS complexes is 99.8%. PMID- 12583266 TI - [A home telemedicine system suitable for China]. AB - The telemedicine developments in china and overseas are introduced in this article. The developmental direction of china home telemedicine, is also discussed, and then a home telemedicine system suitable for china is put forward, and its composition, principle, key components: the home tele-monitor and the remote computer center processing system are introduced in detail. PMID- 12583267 TI - [Study on radiation dose estimation and monitor in TBI using an anthropomorphic phantom]. AB - Absorbed doses and the dose distributions at important tissues and organs in an anthropomorphic phantom are measured using TLD under the TBI conditions. The dose for each tissue or organ is also estimated and monitored for TBI treatment. PMID- 12583268 TI - [The development and application of oxypathor of standard atmosphere]. AB - This paper introduces a newly-developed oxypathor under one absolute atmosphere. It provides both oxygen and mixed-oxygen therapies, featuring oxygen-supply and oxygen-inhalation of hyperbaric oxygen chamber, and broaden the clinical applications of oxygen-therapy, which gave shown satisfactory results. PMID- 12583269 TI - [The development of the hospital plate washer]. AB - The Developments of the Hospital Plate Washer and its control system are introduced in the paper, and the application of the ultrasound technotology in the plate washer is described too. PMID- 12583270 TI - [The development of an electrosurgical apparatus with pulse width control]. AB - This paper introduces an electrosurgical apparatus with pulse width modulating control technique. It mainly consists of pulse width modulation controller, RF power amplifier and coagulation modulator. It makes the RF output power relatively constant. PMID- 12583271 TI - [The amelioration of electrode techniques for testing cerebral potentials]. AB - This article introduces the research work on the new amelioration of electrode techniques for testing cerebral potentials, and the using methods and their advantages & disadvantages in operating the new cerebral electrode. PMID- 12583272 TI - [The status and prospects of X-ray photographs marking technology]. AB - This article introduces three different marking technologies used in clinical radiation photography at present and their respective theories, their advantages and disadvantages, analyzes the space-time and technology conditionality of different marking technologies in selecting mark messages. In the future, new marking technology will be fused with the cassette into one and will contain message as much as possible on photograph marks in order to satisfy the need of technicians and doctors. PMID- 12583273 TI - [Medical X-ray dryview photography and digitized diagnostic technique]. AB - This paper introduces briefly xeroradiography, computed radiography system, x-ray dryview laser imaging series system and its products, digitized diagnostic technique, as well as the electronic radiography etc. PMID- 12583274 TI - [The study about the source, harmfulness and preventive measures of indissoluble particles in injection]. AB - This paper analyzes the source, harmfulness of indissoluble particles in detail and sorts, names them. Some preventive measures for how to reduce the harmfulness are given together. PMID- 12583275 TI - [Dual sieve-mesh collimator]. AB - In this paper is advanced a new type of collimator-dual sieve-mesh collimator. Its principle is to use scanned electronic beam to attack 400 matrix-arranged wolfram targets of the square holes on the upper side of the main collimator and created 400 small X-ray fields sized by 10 mm x 10 mm. These X-ray fields will leave on its center 400 smaller X-ray fields sized by 5 mm x 5 mm after being further collimated by a sub-collimator. What's more, 16 different groups of such X-ray fields can be left on the center of the same field by controlling the all round movement of the main collimator and sub-collimator in proper order and by scanning it round by round and mesh by mesh with electronic beams. It is these ting fields groups that make up various treatment fields with different energy (Dosage). This type of collimator can replace and simplify all other present collimators on accelerators and conformal-intensity modulation radiotherapy equipment. It has advantage of reducing the costs of radio-therapy equipment and making conformal and intensity modulation radiotherapy easier to perform. PMID- 12583276 TI - [The design of measure and control system for composite treating instrument of gastrointestinal cancer]. AB - This paper introduced a composite method of treating gastrointestinal tract cancer-thermo-chemotherapy. A complete set of intelligentialized real-time measure and control system was well designed. The experiment result showed that this system has a stable performance and good real-time. PMID- 12583277 TI - [Study of bone mineral density measuring system with quantitative CT]. AB - This paper introduces a kind of bone mineral density(BMD) measuring system with quantitative CT(QCT). This system includes a self-made solid calibration phantom for BMD and the measuring software. The results of the tests have showed that this system is reliable, cheap and ease of use for BMD measurement. PMID- 12583278 TI - [The study on non-contact detection of breathing and heartbeat based on radar principles]. AB - The paper introduces the design of hardware and software of non-contact detection system for breathing and heartbeat in human body with radar principles and technology. The detection technology is discussed. Under conditions of the illuminating power P < 1 mW and the distance S < 10 m, the non-contact breathing and heartbeat measurement, can be in different positions and with different clothing on the subject. The results show that the system with the technology has a high sensitivity, and is harmless to the health. It is a practicable non contact detection technology for breathing and heartbeat of human body. PMID- 12583279 TI - [A PC-based pulse oximetry monitor for four patients]. AB - Pulse oximeters are used in many departments of a hospital. The pc-based pulse oximetry monitor for many patients has the features of simplicity, flexibility, large memory, powerful function, high ratio of performance to price, and it is easy to communicate with multimonitors. The principles of hardware system and the function of software block of the monitor are introduced too in this paper. PMID- 12583280 TI - [A study on the effects of RF coil for the static magnetic field]. AB - In the paper, we introduce the studies on some abnormal points in static magnetic field, find out the causes and mean while, give the methods of exclusion, thus the adjustment of MRI can be made perfectly. PMID- 12583281 TI - [A viewdata and display system of biologic image by WinDriver]. AB - This paper introduces a viewdata and display system of biologic image by WinDriver, which mainly includes a data acquisition card of image, the ASIC design of its control and an introduction of a method to drive I/O of PC. PMID- 12583282 TI - [The design of ECG sample device based on Windows 98/95]. AB - The ECG sample system which is based on Microsoft Windows98/95 and IBM-compatible PCs ISA bus, is introduced here. It includes ECG sample device, virtual device driver(VxD) and application program, By the method, we can design the device simply and make good use of the computer to programming powerful auto recognition software and telediagnosis software. The system is a cheap and mini medical apparatus. Applications in family care and community medical treatment are encouraging. PMID- 12583283 TI - [The calculation-improving of stress amplification coefficient and the checking of its trouble-shooting model]. AB - Through mechanical analysis, it is discovered that the twined structure decides the amplification coefficient of the catgut tension. Many troubles of production can be explained with this mechanics model, and it could be used as a reference for medical sutures. The paper gives an accurate calculating formula for catgut design and its direct tensile strength through improving mechanics model. It can raise the material strength from 10 to over 31% depending on its twined structure. PMID- 12583284 TI - [Comparison between the artificial cochlea signal processing projects]. AB - The four artificial cochlea signal processors are compared with each other in the paper and a new signal processor system is presented. PMID- 12583285 TI - [The development of an automatic dialyser reuse device by microcomputer]. AB - This article introduces the principles, composition and main functions of an automatic dialyser reuse device by microcomputer. It Ras more practical uses, better reliability, easier operations and greater effect than previous ones, and it's an ideal dialyser ruse device. PMID- 12583286 TI - [The development and mechanism of the argon ion high-frequency combined electrotome]. AB - This article gives you a brief introduction about the main functions and general structure of Argon Ion high-frequency combined electrotome which can reduce hemorrhage in operation, prevent uncontrollable bloold-oozing, shorten the time of operation and decrease the risk of cross-infection. It also recommends some key technologies in the system, especially the use of a computer to monitor and control the system for the security and reliability of the system. PMID- 12583287 TI - [The study advances and prospects of processing surface EMG signal in prosthesis control]. AB - The surface EMG signal has direct relations with extremity movements and has become the ideal control signal for man-machine bionic system. This paper discusses the analytic study of surface EMG signals and gives various character estimations. Two kinds of character recognition methods of surface EMG are introduced for controlling prosthesis movements. The existing problems of multi function EMG prosthesis control are summed up. In the end prospects for the study on analytic ways of surface EMG signal and the practical applications are discussed too. PMID- 12583288 TI - [The development of the multifunctional cephalostat and its applications in anesthesia]. AB - The multifunctional cephalostat is able to hold the patients' head better and firmly at prone position after tracheal intubation under general anesthesia According to the size of patients head, the anesthetist can properly adjust the distance of the malor holder and the distance between the superciliary ridge and the lower jaw, and fix the two tempora to prevent the head swaying. So this instrument greatly facilitates the anesthesia managements. PMID- 12583289 TI - [Acceptance check and quality control of SPECT]. AB - This paper explains the testing of SPECT, especially the new SPECT with double digital detector and spiral scanning frames that has been introduced to China recently, in the acceptance check, proceeding from the physical functions of the system to its mechanical functions, to the NEMA standard functions, and then to the computer hardware specified in the contract. A brief introduction is also given of the quality control of SPECT in terms of its spatial resolution, energy resolution, spatial linearity, sensitivity, and center of rotation. PMID- 12583290 TI - [Study of autocorrelation algorithm and digital accomplishment in ultrasonic medical color flow mapping system]. AB - The theory of autocorrelation algorithm for color flow mapping is analyzed in detail. The digital accomplishment for the algorithm in this paper is also analyzed. With the consistent theory bases, high precision and simply realization, the technology will lay a good foundation for its medical applications in hospitals. PMID- 12583291 TI - [Dual copy-supplement collimator]. AB - Dual Copy-Supplement Collimator consists of two lead conic sections centered on one common radioactive source and having a lot of small square holes on them. When scanning electron beams strike X-Ray target and create some X-Ray fields while passing through the primary section, they penetrate the secondary section and generate even smaller field units(FU). Using these FUs would compose various treatment fields with different energy (dosage). It is able not only to replace many existing collimators such as MLC, but also to make the conformal radiotherapy easier. PMID- 12583292 TI - [The software design for the molecular biological image processing system]. AB - With the use of Visual Basic 6.0 language and molecular biological technology, we have developed a new molecular biological information processing system to detect the characteristic parameter of molecular biological electrophoresis image. The software can determine the molecular weight of the restriction endonuclease hydrolytic DNA fragment and protein or other macromolecules. The application indicates that the system with its accuracy reliability high efficiency and a large capacity of data to be stored, It is very useful to gene diagnosis and molecular biological researches. PMID- 12583293 TI - [PACS-based endoscope image acquisition workstation]. AB - A practical PACS-based Endoscope Image Acquisition Workstation is here introduced. By a Multimedia Video Card, the endoscope video is digitized and captured dynamically or statically into computer. This workstation realizes a variety of functions such as the endoscope video's acquisition and display, as well as the editing, processing, managing, storage, printing, communication of related information. Together with other medical image workstation, it can make up the image sources of PACS for hospitals. In addition, it can also act as an independent endoscopy diagnostic system. PMID- 12583294 TI - [Automatic analyzing system for human chromosome]. AB - A designing method for chromosome automatic analyzing system is presented in the paper. The system accomplishes chromosome automatic recognizing, sorting, pairing, picking out aberrant chromosomes and then makes diseases-diagnosis. PMID- 12583295 TI - [The development of a device producing brain injury by gas percussion]. AB - The device is developed based on aerodynamical principle, measuring technique for feeble signal, and mechano-electronic integration. The left parietals of rats were made cranioectomy and injured by different pressure of gas source with the device and then the survival states, pathological changes and neurobehavioral changes of rats were observed. PMID- 12583296 TI - [The design of a practical instant ECG pocket recorder]. AB - The Holter is no doubt very useful in clinic to help doctor to capture and understand the seizure from the ECG. But the device is not convenient to be use at home and is still very expensive now. This paper introduces a practical instant ECG recorder which a patient can take with at any time. It's cheap and easy of use. PMID- 12583297 TI - [Development of software of the 24-hour dynamic ECG monitoring and analyzing system]. AB - This paper describes a PC-computer program of a digital remote ECG monitoring system. In addition to current monitor's capabilities of real time displaying of dynamic ECG wave, over-limit alarming, and so on, it has developed most functions of the Holter system, that are recording the data of ECG more than 24 hours, reviewing and analyzing the ECG data, and giving out reports. PMID- 12583298 TI - [The development of endoscope workstation]. AB - This paper introduces an endoscope workstation, which solved the weak points of multimedia endoscope database used by most hospitals. The endoscope workstation was built on pedal-switch and NTFS file system. This paper also Introduces how to make program optimal and quick inputting. The workstation has promoted the efficiency of the doctor's operation. PMID- 12583299 TI - [A telemedical multilevel server network system]. AB - Telemedicine is a hot research focus. When we develop the telemedicine system, we find that in China the network condition can not fit the demands of the transregional telemedicine service. In order to improve the poor conditions, we have designed a set of telemedicine system based on distributed multilevel server network model. The paper expounds the structure and the working process of the system, then goes into the technology details in realizing it, finally discusses some relative questions. PMID- 12583300 TI - [The design of a multifunctional mobile operating table]. AB - This article introduces the principles and design of a multifunctional medical table which could be used to immobilize patient from/to bed to/from table, and it can also be used as an operating table. So the dangerous and serious patients can get continuous servo and monitoring during the transfer. The hydraulic pressure lifter, machinery transmission, flat movement and asymmetry structure are employed in the product. PMID- 12583301 TI - [Development and clinical application of the full automatic animal rearing cabin of low oxygen and high carbon dioxide]. AB - This paper introduces a kind of automatic animal rearing cabin of low oxygen and high carbon dioxide. It can mimic the environment of low oxygen and high carbon dioxide at atmospheric pressure and automatically measure and control the concentrations of oxygen and carbon dioxide as well as temperature and humidity in the cabin. The system may provide the equipment support for clinical COPD study. The clinical applications show that the cabin with accurate measurement and control is practical and reliable. PMID- 12583302 TI - [The research and development of computer workstation for Bayer Clinitek urine analyzer]. AB - Computerization and networking is the developing trend of modern medical instruments. This paper describes the research and development work of computer workstation for Bayer Clinitek urine analyzer, including the system designing, hardware composing and software programming. The system can receive analyzing data automatically, manage sample database, print sample report and share data via the network, etc. PMID- 12583303 TI - [The development of an all-purpose medical electronic teaching instrument]. AB - This paper introduces a kind of all-purpose medical electronic teaching instrument. According to the modularization design rule, a medical electronic teaching instrument integrated with information transfer, signal processing, keyboard monitoring and signal sampling, etc, can be realized. On the other hand, A/D, D/A, self-detecting of computer, collecting and testing of biological potentials, driving electromotor and generating functional signals, etc can also be realized by the related software based on the system. It has been proved that, by using single-chip processor and computer, the system can implement full-scale test and processing of biomedical signals so that operators can understand the principles and the construction of medical instruments more easily and deeply. Therefore, it is valuable in application of medical electronic coursesteaching, research and etc. PMID- 12583305 TI - [The design of a new dual-purpose ultraviolet sterilizing light]. AB - This paper introduces the principles, the structure, the working mode of a dual purpose ultraviolet sterilizing light. It is based on the principles of cool and heat air exchanging and laminar flow for sterilization and disinfection. PMID- 12583304 TI - [The study of spuirmy inserted endoscopic robot]. AB - This paper discusses the make-up of the foreing endoscopic robotic system, the model of the moving mechanism, and the application of the squirmy robot. It will provide a new means for minimally invasive surgery, and be valuable to the development of the domestic modern medical engineering. PMID- 12583306 TI - [Lupus nephritis in children and adolescents: clinical and morphologic aspects and clinico-morphologic correlations]. AB - In 53 children and adolescents (47 males, 6 females) with lupus nephritis, clinical features at the time of renal biopsy were analyzed and correlated with pathohistological findings. The mean age at the time of diagnosis of systemic lupus was 12.9 +/- 2.6 (SD) years; the mean ages at the time of diagnosis of lupus nephritis and renal biopsy were 13.5 +/- 2.6 and 13.6 +/- 2.5 years, respectively. The most frequent clinical and laboratory features of lupus nephritis at the time of biopsy were proteinuria (100% of patients), haematuria (88%), nephrotic syndrome (38%), hypertension (32%), and decreased glomerular filtration rate (26%). On initial biopsy, the most frequent finding was diffuse proliferative glomerulonephritis (in 64.1%), followed by "mesangiopathy" (20.8%), minimal changes (7.5%), membranous nephropathy (5.8%), and focal proliferative nephritis (1.9%). Nephrotic syndrome, hypertension, decreased glomerular filtration rate, and haematuria were significantly associated with WHO classes IV and III of lupus nephritis. PMID- 12583307 TI - [Lupus nephritis in children and adolescents: therapy, clinical course and prognosis]. AB - In 53 children and adolescents (47 males, 6 females) with lupus nephritis, clinical features at the time of renal biopsy were analyzed and correlated with pathohistological findings. Therapeutic regimens used and the renal status at the end of follow-up are presented, and factors significantly associated with adverse outcome are analyzed. The mean age at the time of diagnosis of systemic lupus was 12.9 +/- 2.6 (SD) years, the mean ages at the time of diagnosis of lupus nephritis and renal biopsy were 13.5 +/- 2.6 and 13.6 +/- 2.5 years, respectively. The patients having WHO classes I and II of lupus nephritis were treated with prednisone and/or azathioprine; those with WHO classes IV and III were treated with cyclophosphamide (25) or with corticosteroids (5) or with corticosteroids and azathioprine (4), five of them were also treated with plasmapheresis; the patients with WHO class V were treated with prednisone and azathioprine or cyclosporine. Repeated renal biopsies were performed in 12 patients: worsening of morphological lesions was found in four patients, improvement in two and no change was observed in six patients (all with WHO class IV). At the end of follow-up lasting from 0.1 to 14.6 years (mean 4.8 +/- 3.2 years) 80% of patients were in complete (49%) or partial (30%) remission, in 8% of patients the renal disease was clinically active, and in 13% of patients the adverse outcome was noted: one patient died in the first month after diagnosis from extrarenal complications, two patients were in preterminal and the other four in terminal renal failure. Adverse outcome was significantly associated with the presence of nephrotic syndrome at the time of biopsy and with class IV nephritis. The five-year patient's survival rate was 98.1%. The five-year kidney survival rate was 88.6% and 82.4% for the whole group and for the subgroup of patients having classes III and IV nephritis, respectively. PMID- 12583308 TI - [Therapy of lupus nephritis with standard therapeutic protocols and cyclosporine]. AB - The study carried out was at the Department of Nephrology, Military Medical Academy, over the period from 1996 to 2001. Different types of lupus nephritis were documented in 42 patients and were treated with standard therapeutic protocols (corticosteroids, the pulse dose of cyclophosphamide + corticosteroids) and cyclosporine in the target serum concentration of 100-120 ng/ml along with pronisone of 15-20 mg per day. The different degree of damaged renal function was observed. Renal biopsy was performed in 13 patients and in one patient rebiopsy was done. Twenty one patients were treated only with corticosteroids (remission in 23.8% of cases), with cyclophosphamide + corticosteroids 33 patients (remission in 42.4% of cases) and cyclosporine + corticosteroids 12 patients (remission in 91.7% of cases). The pulse therapy with cyclophosphamide in combination with corticosteroids, and cyclosporine in combination with lower doses of corticosteroids was statistically more successful in comparison with corticosteroids monotherapy (p < 0.01). Remission was found in 73.8% of patients, terminal renal weakness was observed in 7 patients, and fatal outcome in 4 patients. We recommend the pulse therapy of cyclophosphamide in combination with corticosteroids in the treatment of severe clinical forms of lupus nephritis, and in refractive forms cyclosporine in combination with low doses of corticosteroids. PMID- 12583309 TI - [Comparison of various cyclophosphamide treatment regimens on the course and outcome of lupus nephritis ]. AB - INTRODUCTION: Immunosuppressive drugs, particularly cyclophosphamide, are widely accepted as the treatment of choice for severe, proliferative lupus nephritis. However, there is no consensus with regard to: 1) the dose required for achieving control of disease activity; 2) duration of cyclophosphamide therapy after the achievement of treatment response; 3) treatment of lupus nephritis relapses [1 5]. In the Institute of Rheumatology, Belgrade, two regimens of intravenous cyclophosphamide have been introduced in the treatment of lupus nephritis patients years ago. The first has comprised the so called "small pulses" that have been used since 1985, and the second has been standard protocol with high doses of cyclophosphamide, accepted in 1990. Results of these follow-up studies were published previously [6-8]. AIM: The aim of this study was to compare the efficacy of two regimens of intravenous pulse cyclophosphamide in the treatment of patients with severe lupus nephritis. METHODS: We analyzed the results of two follow-up studies comprising patients with lupus nephritis, treated with cyclophosphamide: 1) 41 females treated with "small pulses", consisting of 400 mg of cyclophosphamide weekly at treatment onset, followed by the same dose fortnightly for the next three months, and finally on monthly basis for several months or years; 2) 33 patients (29 females and 4 males) treated with standard protocol consisting of "induction phase" with 6 monthly pulses of high doses (0.5 0.75 g/m2 body surface), followed by "maintenance phase" with quarterly pulses for additional 1-2 years. The evaluation of long-term treatment effects was based on remission/response rate [9], number of patients with renal failure, end-stage renal disease and death outcome. RESULTS: Groups of patients were quite comparable with respect to their demographic and clinical data (Table 1). The only difference was much higher frequency of renal biopsy in "high dose" cyclophosphamide pulse (85% versus 32%), confirming the presence of proliferative lupus nephritis. Cummulative dose of cyclophosphamide and treatment duration were not significantly different between treatment groups. At the end of the follow up, distributions of favorable (remission/response) and unfavorable outcome was similar (p = 0.831; Mann-Whitney U test), as well as dynamics of remission achieving (p = 0.068; Log-rank test), cummulative renal survival (p = 0.129; Log rank test) and patient survival (p = 0.577; Log-rank test). DISCUSSION: Similar efficacy of two different cyclophosphamide regimens in our patients with lupus nephritis was not surprising considering that cummulative cyclophosphamide doses and treatment duration were similar obtaining similar control of disease. During induction phase of treatment, patients on small pulses have received even higher cummulative dose of cyclophosphamide. Aggressive immunosuppressive treatment with cyclophosphamide has significantly ameliorated the outcome of lupus nephritis. In different studies, rate of assessed clinical response is 60-80 [13-17]. Significant proportion (42%) of patients who achieved partial remission, as well as complete remission, developed flare of renal disease several months after the end of the treatment, necessitating restarting of pulse cyclophosphamide therapy. The results of our study were in accordance with those results, especially with results of Mosca et al. [18] who have applied the duration of treatment similar to ours in high pulse regimen. CONCLUSION: Treatment response did not differ between two different cyclophosphamide regimens (small pulses and standard high doses protocol), but standard protocol seemed to be more comfortable for patients. We recommend standard protocol for patients with biopsy proved proliferative lupus nephritis as a gold treatment standard. However, sustained remission of proliferative lupus nephritis is a goal that still remains to be achieved. PMID- 12583310 TI - [Lupus nephritis: retrospective analysis of the course of the disease]. AB - INTRODUCTION: Systemic lupus erythematosus is a multisystemic autoimmune disease characterized by various clinical manifestations, as well as unpredictable course and therapy response. Lupus nephritis is a severe manifestation of the disease, significantly influencing its prognosis. Since the pathogenesis of lupus nephritis has not been elucidated, optimal therapeutic approach is still subject of numerous studies. AIM: A retrospective study was undertaken with the aim to analyze clinical features of different types of lupus nephritis, their therapy response as well as their course and outcome. PATIENTS AND METHODS: The retrospective study included 67 patients who were regularly followed-up for 36 188 months after renal biopsy at the Department of Nephrology, Clinical Centre of Serbia or at the Department of Nephrology, Clinical Centre of Montenegro. Systemic lupus erythematosus was diagnosed according to the ARA criteria, while pathomorphological classification was based on WHO recommendations. RESULTS: The studied group was mostly composed of patients with proliferative lupus nephritis (21 with diffuse and 9 with focal) followed by 20 patients with membranous, 14 with mesangioproliferative (type IIb) and only one patient with type I, IIa and VI lupus nephritis each. Age distribution of the patients and incidence of extrarenal complications was equal in patient groups with different types of lupus nephritis. Only four patients were males. Choice of therapy depended on the morphological characteristics of lupus nephritis, presence and severity of extrarenal complications and renal function. At the time of analysis, 10/14 patients with lupus nephritis type IIb were in remission, as well as 14/30 type III/IV patients and 15/20 patients with type V lupus nephritis. The outcome was the worst in patients with proliferative lupus nephritis, and thus, at the time of analysis four of 30 patients had chronic renal failure, 7 were on regular haemodialysis, while 3 patients decreased. With respect to the renal function, the most favorable outcome was recorded in IIb patients, while mortality was not dependent on lupus nephritis type but on extrarenal manifestations of systemic lupus and associated diseases. Five- and ten-year patients' survival rates were 100% and 86%, respectively. Cardiovascular and cerebrovascular diseases were the most frequent causes of death. CONCLUSION: Persistent and controlled treatment using a combination of different immunosuppressive drugs along with close follow up of patients and possibility of dialysis treatment have significantly improved prognosis of patients with lupus nephritis, even in those with diffuse proliferative changes. PMID- 12583311 TI - [Long term survival of patients with lupus nephritis]. AB - Lupus nephritis is a clinical manifestation of Systemic Lupus Erythematosus with most prominent influence on the course of the disease. The most predictive parameters for development of renal failure are: type of hystological changes, degree of interstitial inflammation, serum creatinine concentration at the time of diagnosis and therapeutical protocols used in the treatment. Single center experience in a group of 220 lupus patients is presented in this paper. In 130 patients (59%) lupus nephritis was diagnosed by clinical and laboratory tests, while 74 kidney biopsies were performed. Proliferative type of lupus nephritis (class IV in 54% of cases and class III in 18.9%) was more frequent than the other histological types. During a long term follow up (range 1-17 years, mean 7.6 years) renal failure developed in 17 patients (24%), while 11% of patients developed uremia and required dialysis. Development of renal failure was influenced by histological changes with predominance of class IV lupus nephropathy, disseminated disease with more ARA criteria for systemic lupus erythematosus classification present at the time of kidney biopsy, more frequent lupus flares, persistence of low complement levels during the first year of follow-up and even earlier. PMID- 12583312 TI - [Laboratory and morphologic parameters in patients with lupus nephritis]. AB - INTRODUCTION: Lupus nephritis is an example of glomerulonephritis mediated by immune complexes. The information obtained by kidney biopsy corroborates diagnosis and evaluation of disease activity, specify of prognosis and mode of treatment. The object of our study was to determine the prevalence of particular pathohistological types of lupus nephritis in our group of patients, to establish if there was a correlation of laboratory and morphological parameters, and to present the use of specific therapeutical protocols. METHOD: The study included 58 patients with diagnosed systemic lupus erythematosus (SLE) and lupus nephritis, who had biopsy of kidneys. The indications for biopsy were the following: proteinuria level over 0.5 g/24 hrs, erythrocyturia and cylindruria. The patients were examined and treated at the Institute of Allergology and Immunology, Clinical Centre in Belgrade, over the period 1994-2001. Within the testing, besides standard laboratory tests, the immunological evaluation was also performed as follows: the level of standard serum immunoglobulins, C3 and C4 components of complement, antinuclear antibodies (ANA) and antibodies to double stranded DNA (dsDNA), were determined. RESULTS: There was 84.48% of female patients in the studied group. The mean-age was 36.5 years, while the average duration of disease (SLE) to kidney biopsy was 28.3 months. Considering cytopenia, leukopenia was found in 26.79% of patients, lymphopenia was recorded in 62.26% of cases while anaemia was noted in 52.63% of patients. The values of serum creatinine were elevated in 25.86% of patients, while creatinine clearance rate was lower (below 80 ml/min) in 75% of cases. The values of proteinuria are illustrated in Graph I. Cylindruria was found in 20.69% of subjects, massive erythrocyturia in 44.83%, and 46.55% of patients had more than 5 red blood cells in urinary sediment. Regarding the pathohistological findings, according to WHO classification, the biopsy of kidneys revealed the following distribution: class I--3.45% of patients, class IIA--24.14%, class IIB--31.03%, class III--12.07%, class IV--24.14%, class V--3.35%, and class VI 1.72% of cases. DISCUSSION: Within the immunological evaluation, the increased serum immunoglobulin G (IgG) level was found in 26.79% of subjects, suggesting that the consumption of complements in formation of immune complexes was the basic pathogenetic mechanism of lupus nephritis. Positive finding of ANA was recorded in about 95% of subjects, what was typical for SLE, while antibodies to dsDNA were positive in no less than 72.72% of cases, arguing for the fact that they were one of major nephritogenic antibodies. Considering the correlation analysis, no correlation between pathohistological findings and serum creatinine level was found, but there was the correlation between pathohistological findings and decreased creatinine clearance rate. The correlation between pathohistological findings and proteinuria up to 0.5 g/24 hrs was verified. There was no correlation between the increased IgG level and kidney biopsy findings, but it was found that decreased level of C4 complement component correlated with the degree of kidney lesion. The value of diastolic pressure also correlated with pathohistological findings. Concerning the applied mode of treatment, 46.55% of patients were administered pulse doses of cyclophosphamide combined with pulse doses of methylprednisolone, 5.17% had pulse doses of cyclophosphamide and 32.76% pulse doses of methylprednisolone, while 12.07% received glucocorticoid drugs combined with azathioprine, and glucocorticoids only were given to 3.45% of them. The conclusion will be that the biopsy of kidneys is the imperative in the evaluation of lupus nephritis, because the complete insight into the degree and type of kidney lesion as well as search for an optimal mode of treatment may be achieved only by assessment of combined clinical, laboratory and morphological parameters. PMID- 12583313 TI - [The importance of renal biopsy in lupus nephritis]. PMID- 12583314 TI - [Recommendations for the treatment of lupus nephritis]. PMID- 12583315 TI - [Conservative therapy of inflammatory bowel diseases]. AB - In the past years there have been many exciting developments in IBD management. New therapies and concepts of remission induction and of maintenance have been developed. This paper is intending to bring together the basic topics in patient management in the various medical fields that represent the most accepted ways of therapies. The introduction deals with the most common aspects of Crohn's disease and that of the ulcerative colitis. It describes the treatment of ulcerative colitis according to the different groups of patients and the degree of activity. First it analyses the possible ways leading to the remission, then the prevention of relapse, lastly the other therapeutic options which have not been given any evidence of so far. The main standards of Crohn's disease therapy are presented on the bases of the small bowel Crohn's disease. After discussing the essential principles in treating the complications it carries on the treating of the large bowel Crohn's disease and then it is completed with the consideration of prevention of the relapse and the supportive therapeutic possibilities. PMID- 12583316 TI - [Femoral press-fit fixation technique in anterior cruciate ligament reconstruction. Preliminary report]. AB - The authors have reported the experiences at femoral press-fit fixation technique used in anterior cruciate ligament reconstruction with bone-patellar tendon-bone autograft. Their surgical technique have been described and early results have been compared with those from anterior cruciate ligament plasties with the interference screw fixation technique during a prospective study. PATIENTS AND METHODS: 665 anterior cruciate ligament reconstructions were performed between September 1997 and August 1999 at the Department of Sports Surgery of the National Institute for Sports Medicine. 418 of them were operated on by transtibial technique using free bone--patellar tendon--bone autografts with 265 femoral press-fit and 153 interference screw fixations. 110 patients from the press-fit and 72 from the interference screw group could be followed-up (average follow-up period: 19 months). Physical examination, evaluation by International Knee Documentation Committee and Lysholm scores, radiology, and stability test by KT-1000 arthrometer were used as follow-up tests with statistical comparison. RESULTS: According to the International Knee Documentation Committee score, 77% of the patients with the interference screw and 87% of the patients from the press-fit group turned out to get to the normal or nearly normal categories. Neither the Lachman, anterior drawer and pivot shift tests, nor the results of the KT-1000 measurements have shown statistically significant difference between the two groups of patients. Mild (3-5 degrees) extension deficit has been detected at 14% of the patients with interference screw and 8% of the patients in the press-fit group. Mild (6-15 degrees) flexion deficit has been observed at 22% of the patients with interference screw and 24% of the patients from press-fit group with moderate (16-25 degrees) of flexion deficit at 4% of the patients. The subjective opinions of more than 90% of the patients were that the operation proved to be excellent or good. Based on the functional and stability data of the follow-up study, 87% of the patients operated on with femoral press-fit autograft fixation technique for their anterior cruciate ligament have shown excellent or good results. CONCLUSIONS: When compared with the interference screw technique well-accepted in the literature, no statistically significant difference could be found, therefore this method can be recommended as alternative one for the femoral fixation of the anterior cruciate ligament graft. PMID- 12583318 TI - [Evaluation of fitness to drive--conclusions after a case report]. AB - Health evaluation of candidates applying driving license is general practitioner's duty, in the case of driving private cars. In Hungary no generally accepted guideline available on the evaluation process, the necessary examinations etc. A patient with a III degree A-V block was treated by the authors after suffering a serious syncope, with brain contusion, however he was regarded fit to drive by his GP, after the serious accident. The patient after the brain trauma was treated in two different institutions, however no ECG record was found, when the authors asked it to find out the time of the A-V block. Finally the patients was treated with temporary and later on with permanent pace maker. During the half year follow up the patient was symptom less. The authors summarize the possible mechanism of syncope, and the criteria the evaluation of patients with heart diseases. In conclusion the authors emphasize the need for a generally accepted Hungarian guideline in the evaluation process of candidates applying driving license. PMID- 12583317 TI - [Detection of trisomy 8 and monosomy 7 in chronic granulocytic leukemia and myelodysplastic syndrome by cytogenetic analysis and fluorescence in situ hybridization]. AB - INTRODUCTION AND AIMS: In this retrospective study the authors studied how interphase fluorescence in situ hybridization can be applied to the diagnosis of numerical chromosomal anomalies failed to be identified by cytogenetic analysis. METHODS: Thirty-four patients, 27 with chronic granulocytic leukemia and seven with myelodysplastic syndrome, were studied in order to identify disease-specific aberrations, trisomy 8 and monosomy 7 using both traditional cytogenetic analysis and fluorescence in situ hybridization on interphase bone marrow cells. Using alphoid-satellite centromeric specific probes, trisomy 8 indicating the progression of the disease and poor prognosis in chronic granulocytic leukemia as well as monosomy 7 in myelodysplastic syndrome were identified. RESULTS: In 21 of 34 cases both methods led to the same results. In 13 patients fluorescence in situ hybridization making possible to examine a great number of cells, provided more information about the gain and loss of chromosomes above and clarified uncertain cytogenetic results. CONCLUSIONS: In both hematological malignancies, studied by the authors, fluorescence in situ hybridization proved a useful and sensitive method to determine chromosomes unrecognized or not accurately identified by the traditional cytogenetic analysis and to define the ratio of pathological cells. At the same time the results confirm that conventional cytogenetic analysis is still essential in diagnosing the genetic alterations of malignant cells and point at the chromosomes that are worth further studying by other methods. PMID- 12583319 TI - [Clinical electrocardiography is 100 years old]. PMID- 12583320 TI - [Clinical aspects of thyroid diseases. 1952]. PMID- 12583321 TI - [Contribution to the problem of atrial fibrillation and anticoagulant treatment]. PMID- 12583322 TI - [Fibrinolysis modulators]. AB - The structure of fibrin network and the properties of the fibrinolytic enzymes (the formation, the efficiency of their activity and the rate of their inactivation) essentially determine the effectiveness of fibrinolysis. The fibrin structure and the action of proteases, however, depend considerably on additional physiological and pathological conditions. These are: cell membrane and components of endothelial cell, elastase of polymorphonuclear cells, actin and myosin in thrombus released by platelets and smooth muscle cells, certain denatured proteins, various immunoglobulins, some metabolites accumulated in diabetes mellitus, the inhibitor system of fibrinolytic enzymes (where and when proteases are available for inhibitors). In this survey, the biochemical basis makes understandable for the medical practitioner why urgent fibrinolytic therapy is required and why it would be ideal to initiate the formation of fibrinolytic enzymes at the site of thrombus. PMID- 12583323 TI - [Diabetes specialist care at the turn of the Millennium in Bekes County of Hungary (experience with 5935 new patients between 1996 and 2000)]. AB - INTRODUCTION: Authors collect systematically the main demographic data of the registered new diabetic patients, who are presented in the specialist diabetes care station of county Bekes. AIM(S): These data are compared with the similar data of the previous five years periods. PATIENTS: According to the present sixth registration, the number of the new diabetic patients increased rather (5935 patients), while the inhabitants of county Bekes decreased. RESULTS: The proportions as per age and gender did not change in comparison with the earlier periods, but in some age-decade male predominance could be observed. Recently most of the patients are those over 50 years (type 2) with the striking increase over 70 years, too. The number of obese patients is constantly high. The ratios of employments are considerably changed vs the first examined period (before 25 years), the number of retired patients is increased and that of household women is decreased. Proportion of the different therapeutic regimens is near to what is required (reaching of normoglycaemia), but the number of patients treated with the oral antidiabetics is somewhat higher as desired. Number of those needing insulin is increasing. CONCLUSION: Recently (31. Dec. 2000) altogether 30.446 new diabetic patients are registered in county Bekes, the incidence in the last period is 30,44/10.000 subjects/year. PMID- 12583324 TI - [Role of the TNF system in the pathomechanism of bronchial asthma]. AB - INTRODUCTION: The activation of the TNF system during asthmatic attack has already been proved. AIMS: The aim of the study was to reveal the role of the tumor necrosis factor (TNF) system in the pathomechanism of bronchial asthma. PATIENTS/METHODS: Childhood asthmatic lately symptom-free adults (n:39) and their non-asthmatic offspring were examined (n:42). According to the methacholin airway challenge test, patients were divided into bronchial hyperreactive (n:44) and non hyperreactive (n:37) groups. Tumor necrosis factor alpha (TNF-alpha) and its soluble receptors 55 (sTNF-R1) and 75 kDa (sTNF-R2) were measured by ELISA. RESULTS: Among the hyperreactive patients (n:44) significantly higher TNF-alpha (mean +/- SD: 5.13 +/- 1.37 vs. 3.91 +/- 0.61 pg/ml, p < 0.0001), sTNF-R1 (mean +/- SD: 1.37 +/- 0.28 vs. 1.16 +/- 0.13 ng/ml, p = 0.0002) and sTNF-R2 (mean +/- SD: 0.78 +/- 0.42 vs. 0.43 +/- 0.41 ng/ml, p < 0.0001) values were measured compared to the non-hyperreactives (n:37). In hyperreactive patients there was a significant correlation between the cytokine and cytokine receptor levels (TNF alpha-sTNF-R1 p = 0.0184, r = 0.3541; TNF-alpha-sTNF-R2 p < 0.0001, r = 0.6468). Significant negative correlation was detected between the serum TNF-alpha and sTNF-R2 concentrations and PD20 FEV1 methacholin (dose of methacholin resulting in a 20% reduction of forced exspiratory volume in 1 second) in hyperreactive patients. CONCLUSION: According to our results the activation of the TNF system may contribute to the bronchial hyperreactivity. It can be observed in asthmatic patients having been symptom-free for years and in their non-asthmatic offspring as well. These results refer to the presence of a minimal allergic inflammation. PMID- 12583325 TI - [Safety profile of NO-SPA]. AB - The aim of the safety analysis performed by Chinoin Drug Safety Unit was to summarise the safety profile of NO-SPA (drotaverine hydrochloride), the Hungarian spasmolytic well known in Hungary and abroad. Authors collected the safety data from clinical studies between 1964-1998 for the determination of the adverse event frequency. Based on the data of 12111 patients treated with NO-SPA in 37 clinical trials 0.9% frequency of adverse events was found. The value indicates uncommon (0.1-1%) adverse event frequency according to the criteria for frequency categories. The benefit-risk ratio of NO-SPA is favourable, since the therapeutic effect does not include frequent adverse reaction occurrence. PMID- 12583326 TI - [Maternal toxoplasma infection and mosaic trisomy 8 syndrome]. AB - INTRODUCTION: Mosaic trisomy 8 syndrome shows significant phenotypic variability. High forehead, deep set eyes, wide base of the nose, upturned tip of the nose, protruding lips, prominent ears, strabism, corpus callosum agenesia, deep plantar furrow and variable degree of psychomotor retardation are present in most patients. PATIENTS/METHODS: The authors describe a patient with severe motor and mental retardation, optic disc hypoplasia, cardiomyopathy and partial extrahepatic bile-duct atresia, who's mother was treated for toxoplasma infection in her pregnancy. Perinatal infection of the child was ruled out by the documentation of decreasing IgG values in the mother and newborn and normal IgM levels in the newborn. RESULTS: Facial characteristics and the bilateral deep plantar furrow raised the possibility of trisomy 8 syndrome. Routine chromosomal analysis and fluorescent in situ hybridization studies revealed a normal cell line, mosaic trisomy 8 in 13% of the cells, and a pericentric marker 8 ring chromosome in 27% of the cells. CONCLUSION: The authors suggest that the periconceptional toxoplasma infection may play a role in the occurrence of the mosaic trisomy presenting with cardiomyopathy and partial bile-duct atresia previously undescribed in this syndrome. PMID- 12583327 TI - [Study on the technique of Rehmannia glutinosa virus-free]. AB - The stem tip from germ-free stem segment of Rehmannia glutinosa cultured in test tube can be induced into virus-free seedling. The experiment showed that the proper disinfectant for stem segments of Rehmannia glutinosa was 0.05% HgCl2. The seedings from stem segments grew better with MS in the concentration of agar 0.7% and pH7. The stem tips could be directly induced to seedlings by using MS + 6 BA(0.05 mg/L). The MS media for seedlings virus-free culture are 1/4 macro elements + 1/2 micro-elements and using edible sugar instead of sucrose, so the cost of media could be decreased to 48.7%. PMID- 12583328 TI - [Effect of lanthanum on lead absorption in soybean seedling]. AB - The rare elements may have positive effects on growth of plants, and they are the beneficial elements to plants. Lead is one of heavy metals which are harmful to plants and human body. In this paper, the effect of lanthanum on lead absorption in soybean seedling were researched by water culture. Lead in soybean seedling was measured by AAS. The experimental results showed that the level of lead in soybean increased with an increase in the concentration of lead in cultural medium, while the level of lead in soybean reduced after adding La to the cultural solution. PMID- 12583329 TI - [Microscopy identification of Phyllanthus simplex]. AB - OBJECTIVE: To identify Phyllanthus simplex. METHODS: Morphological and histological identification. RESULTS: There are cluster crystals in cortex and mesophyll of the root and the stem from Phyllanthus simplex. Paracytic type stomata are visible. CONCLUSION: Above-mentioned characters can be used for the identification of Phyllanthus simplex. PMID- 12583330 TI - [Identification of Ligesticum chuanxiong by X-ray diffraction Fourier pattern method]. AB - OBJECTIVE: To establish a new identification and analysis method of Chinese medicinal materia Ligesticum chuanxiong Hort. METHODS: Powder X-ray diffraction Fourier pattern. RESULTS: Experiments and analysis were carried out on five samples. The standard X-ray diffraction Fourier pattern and characteristic diffraction peaks of Ligesticum chuanxiong Hort. were obtained. CONCLUSION: This method can be used for identification of Ligesticum chuanxiong Hort. PMID- 12583331 TI - [Descriptions and microscopy identification of Citrus grandis var. tomentosa Hort. and Citrus grandis Osbeck]. AB - OBJECTIVE: To identify Citrus grandis var. tomentosa Hort. and Citrus grandis Osbeck. METHODS: Descriptions and microscopy identification. RESULTS: There are nonglandular hair in the stem epidermis, many starch grains in the stem cortex and many prismatic crystals of calcium oxalate in the phloem of C. grandis var. tomentosa Hart. The palisade tissues of the leave are thick. The outer layer palisade tissues contains prismatic crystals. The cortex of leaf vein is thick. The other characters are similar to C. grandis Osbeck. CONCLUSION: Above mentioned characters can be used to identify C. grandis var. tomentosa Hort. and C. grandis Osbeck. PMID- 12583332 TI - [Study on the biological assay of Herba Leonuri--analysis the dosage response curve of Herba Leonuri and oxytocin and establishment of adequate potency pattern]. AB - OBJECTIVE: To establish the effective patten of Herba Leonuri to be suitable for biological assay. METHODS: By comparing the dosage-effect curve of Herba Leonuri and oxytocin the suitable effective pattern was set up. RESULTS: The experiment suggest that the uterus contract height from 20 mm to 85 mm is the adequate effective range. In this range, the effect both Herba Leonuri and oxytocin should plot as a straight line against the log dose and two lines are basically parallel. CONCLUSION: In the light of establishing effective pattern, the biological assay of Herba Leonuri may pattern the method of oxytocin biological assay to regard oxytocin as standard. The result error of Herba Leonuri assay is under 20%, not exceed the demand limits of biological assay. PMID- 12583333 TI - [Study on the permeability of 125I-sinomenine via the transdermal delivery way in rats]. AB - OBJECTIVE: To study the permeance and diffusion trend of 125I-sinomenine via the transdermal delivery way in rats. METHODS: Assay the permeance depth and diffusion area of sinomenine labeled by isotopic 125I in subcutaneous of rats certain time after sinomenine was administrated on the cervical skin of rats. RESULTS: The diffuse of sionmenine can reach cervical spine in longitudinal direction and form an area of 2 cm2 in transverse direction. The concentration gradient of both directions are significant. CONCLUSION: 125I-sinomenine can not only be absorbed into blood via the transdermal delivery way to have whole effect, but also accumulate large amounts in the skin to have a local effect. PMID- 12583334 TI - [Experimental study on anti-thrombus effect of different extracts from Semen Persicae]. AB - OBJECTIVE: To observe the effects of anti-thrombus of different extracts from Semen Persicae and provide experimental basis for finding and defining effective fraction of Semen Persicae. METHODS: The effects of different extracts from Semen Persicae on clotting time in mice, pulmonary embolism in mice and electro irritate thrombosis in rats were observed. RESULT: Ethyl acetate extract from Semen Persicae could prolong clotting time in mice, markedly relieve respiratory difficult due to pulmonary embolism in mice induced by ADP-Na, obviously retard the generation of thrombosis induced by electrical injury of carotid artery in rats. CONCLUSION: Ethyl acetate extract from Semen Persicae possesses evidently anti-thrombotic effect. PMID- 12583335 TI - [Effect of changtong oral liquids on the fibrinolytic activity of rabbits with experimental intestinal adhension]. AB - OBJECTIVE: To observe the effect of Changtong Oral Liquid (CTOL) on the fibrinolytic activity of rabbits with experimental intestinal adhesion. METHODS: Intestinal adhesion model of rabbits were made with reference to that described by Thomas and Rhoads and the animals were continuously orally supplemented with CTOL for seven days. Changes of tissue plasminogen activator (t-pA) and plasminogen activator inhibitor activity (PAI) as well as the intestinal adhesion were investigated. RESULTS: Different dosage of CTOL produced protective effects on the postoperative intestinal adhesion accompanied by significant enhancement of t-PA activity and pronounced decreasement of PAI activity. CONCLUSION: CTOL maybe play protective role in postoperative intestinal adhesion by modulating the activity of fibrinolytic system. PMID- 12583336 TI - [The pharmacological studies of yifuzhixue pill]. AB - OBJECTIVE: To investigate the pharmacological effect of Yifuzhixue Pill (YFZXP). METHODS: PPT, RT, PT, platelet counts and platelet adhesive rate were examined after administration of YFZXP. In other hand, the functions of isolated rabbit's uteri were also examined to determine whether they were responsible to the administration of YFZXP. RESULTS: YFZXP could enhance the intrinsic blood coagulating function and stimulate the contraction of smooth muscles of uterus including increasing the rhythms and strength. The stimulation effect of YFZXP on smooth muscles of uterus exhibited no selections between the uteri treated with or without estradiol. It was speculated that the increased intrinsic blood coagulating and uterus' contraction induced by YFZXP were contributed to its anti bleeding effect in uterus. PMID- 12583337 TI - [Hypoglycemic effect of saponin from Tribulus terrestris]. AB - OBJECTIVE: To study the hypoglycemic effect of saponin from Tribulus terrestris L. METHOD: Alloxan was used to establish the diabetic model in mice. Phenformin Hydrochlride Tables was used as the positive control. The level of glucose, triglyceride, cholesterol and SOD in serum were determined. RESULTS: The level of serum glucose could be significantly reduced by saponin from Tribulus terrestris, which was the rate of 26.25% and 40.67% in normal mice and diabetic mice in respectively. The level of serum triglyceride could be reduced 23.35%. The saporin could also decrease the content of serum cholesterol. Serum SOD activity of the mice was increased by the saponin. CONCLUSION: Saponin from Tribulue terrestris could significantly reduce the level of serum glucose. PMID- 12583338 TI - [Long-term toxicity experiment with tea for descending lipid and declining fat]. AB - OBJECTIVE: To investigate the chronic toxicity of tea for descending lipid and declining fat (TDLDF). METHODS: 4 groups Sprague-Dawley rats had respectively been fed with distilled water and TDLDF in 7 g/kg (high dosage), 3.2 g/kg (middle dosage) and 1.5 g/kg(lower dosage) for 12 weeks, the latter were respectively equal to 100, 50 and 20 times clinical dosage. After ig 6, 12 weeks and 3 weeks after discontinuation, the behavior, appetite, excrement and urine, body weight, hematological and biochemical markers, coefficient of the main organs and pathohistological examination of the rats were investigated. RESULT: There were no significant differences of the above markers between each experimental group and the control group. CONCLUSION: The clinical recommended dosage of TDLDF is safe. PMID- 12583339 TI - [Experimental study on the hypoglycemic effect of tangniaole capsule]. AB - OBJECTIVE: To study on the hypoglycemic effect of Tangniaole Capsule. METHOD: The effects of Tangniaole Capsule were proved through experiments on both normal and diabetic animals. RESULTS: Tangniaole Capsule was able to decrease the blood sugar content in normal mice and rats, heighten the level of serum insulin and the content of liver glycogen in normal rats, decrease the blood-sugar content and increase the content of liver glycogen in hyperglycemina rats induced by adrenalin, inhibit the alloxan-induced hyperglycemia of mice and rats and improve the glucose tolerance of rats. CONCLUSION: Tangniaole Capsule have obvious hypoglycemic effect. PMID- 12583340 TI - [Studies on extraction process of Radix Platycodi]. AB - The orthogonal design was used to optimize the extraction process of Radix Platycodi with content of total saponin and yield of the extract as markers. Factors that have been chosen were alcohol concentration, alcohol consumption, extraction times and extraction time. Each factor has three levels. The result showed that the optimum extraction condition obtained was 70% alcohol, 3 times the amount of material, refluxing for 5 times, 60 minutes each time, the optimized process was stable and workable. PMID- 12583341 TI - [The effect of he jie decoction on the TCRV beta 7 of chronic hepatitis B patients]. AB - OBJECTIVE: To explore the correlation of clinical effect and TCRV beta 7 expression of He Jie Decoction (HJD) on chronic hepatitis B patients. METHODS: 45 patients of chronic hepatitis B were divided randomly into two groups. 30 patients in the treated group were treated by HJD, and 15 patients in the control group were treated by usual western medicine. The TCRV beta 7 level were detected before and after being treated. RESULTS: After six months treatment, the ALT level of two groups were obviously decreased (P < 0.01), TCRV beta 7 expression were detected out in 6 patients of the treated group, and HBV-DNA and HBeAg of the 6 patients were negative conversion. The TCRV beta 7 expression couldn't be detected out in the control group, and HBV-DNA and HBeAg of the control group weren't negative conversion. There was no significant difference between the clinical total effective rate of the two groups (P > 0.05), but the apparent rate of the treated group was significantly higher than that of the control group (P < 0.01). CONCLUSION: HJD might have the effect on improving the TCRV beta 7 expression of chronic hepatitis B patients, which might be the ways of HJD inhibiting and killing hepatitis B virus. PMID- 12583342 TI - [Sacer ignis, quam pustulam vocant pastores: anthrax--cultural historical traces of a zoonosis]. AB - The knowledge of anthrax as a disease and its importance as a zoonosis in the Greco-Roman world is revealed through a selection of classical texts and mythological sources, taking into account evidence of reworking and reuse of these texts up until the nineteenth century. The numerous names given to the disease throughout history and their linguistic origins will also be examined in this paper. The narrative of the epizoonoses in Noricum in Virgil's Georgics; taken by several to represent a description of an anthrax epidemic, and which had a great influence in written works on veterinary medicine up until the discovery of bacteria, will be given particular attention. The crucial term is "Sacer Ignis", synonymous for several different human and animal diseases through time. This term will be analysed in terms of linguistic origin and the changes in meaning it acquired throughout the centuries. PMID- 12583343 TI - [The Veterinary Society of Fribourg and Martin Strebel]. PMID- 12583344 TI - [The two Swiss veterinary schools: their varied history between equality and merger]. PMID- 12583345 TI - [Veterinary medicine of the GDR in strained relations between technical instructions and political guidelines]. AB - The development of Veterinary Medicine in the Soviet Occupation Zone in Germany and the former German Democratic Republic (GDR) is sketched in highlights. After the collectivization of agriculture (1960) a centralistically controlled national veterinary system was established. It was suited to the requirements of the industrially organized animal production. The successive classification of socialistic veterinary administration was associated with the following matters: the extension of veterinary subject matters, a vertical division of work with the aid of newly created technical veterinary professions, and a penetration of the profession with political guidelines. As the professional level of the veterinary system in the GDR was relatively high the reflection in retrospective needs to be evaluated in a differentiated way considering the textual and social conditions. In spite of centralism and indoctrination the veterinary system remained professionally autonomous with islands of political independence, which sustained the identity of this profession. The latter formed the base for self renewal of the East German veterinary system at the end of the socialistic area 1989/1990. PMID- 12583346 TI - Mutation patterns of mitochondrial H- and L-strand DNA in closely related Cyprinid fishes. AB - Mitochondrial genome replication is asymmetric. Replication starts from the origin of heavy (H)-strand replication, displacing the parental H-strand as it proceeds along the molecule. The H-strand remains single stranded until light (L) strand replication is initiated from a second origin of replication. It has been suggested that single-stranded H-strand DNA is more sensitive to mutational damage, giving rise to substitutional rate differences between the two strands and among genes in mammalian mitochondrial DNA. In this study, we analyzed sequences of the cytochrome b, ND4, ND4L, and COI genes of cyprinid fishes to investigate rates and patterns of nucleotide substitution in the mitochondrial genome. To test for strand-asymmetric mutation pressure, a likelihood-ratio test was developed and applied to the cyprinid sequences. Patterns of substitution and levels of strand-asymmetric mutation pressure were largely consistent with a mutation gradient between the H- and L-strand origins of replication. Significant strand bias was observed among rates of transitional substitution. However, biological interpretation of the direction and strength of strand asymmetry for specific classes of substitutions is problematic. The problem occurs because the rate of any single class of substitution inferred from one strand is actually a sum of rates on two strands. The validity of the likelihood-ratio test is not affected by this problem. PMID- 12583347 TI - Uninsurance rates vary widely across communities and regions. PMID- 12583348 TI - More small firms offer health insurance but fewer employees enroll. PMID- 12583349 TI - Tracking health care costs: an upswing in premiums and costs underlying health insurance. AB - This Data Bulletin is based on data from the Milliman & Robertson Health Cost Index, which is designed to reflect cost increases faced by private insurers; the Kaiser Family Foundation/Hospital Research and Education Trust survey of employer based health plans for 1998-2000; and the KPMG survey of employer-based health plans for 1993-1998. PMID- 12583350 TI - [Advising the expecting mother on the use of epidural anaesthetics in obstetrics]. PMID- 12583351 TI - [The safety and registration of pharmaceuticals from the point of view of the medical practitioner]. PMID- 12583352 TI - Transforming growth factor-beta levels in synovial fluid of osteoarthritis with or without calcium pyrophosphate dihydrate crystals. PMID- 12583353 TI - Cardiovascular thrombotic events and COX-2 inhibitors: results in patients with osteoarthritis receiving rofecoxib. PMID- 12583354 TI - Diabetes mellitus type 1 and stiff hips. PMID- 12583355 TI - The California Tobacco Control Program and potential harm reduction through reduced cigarette consumption in continuing smokers. AB - Harm reduction for continuing smokers has been suggested as a public health priority. We evaluated whether tobacco control programs might reduce cigarette consumption among current smokers through strategies aimed primarily at protecting nonsmokers from secondhand smoke (SHS). Data were from adult (18+ years) respondents to multiple (1990, 1992, 1996, 1999), large, cross-sectional, population-based surveys of smoking behavior, conducted to evaluate the California Tobacco Control Program. Adult daily smoking prevalence decreased from 15.9 +/-0.4%(+/-95% confidence interval) of the California adult population in 1990 to 13.0 +/-0.3% in 1999. Concurrently, moderate-to-heavy daily smoking (>or= 15 cigarettes/day) decreased from 10.3 +/-0.4% in 1990 to 7.4 +/-0.3% in 1999, and heavy daily smoking (>or= 25 cigarettes/day) from 3.4 +/- 0.2% in 1990 to 1.9 +/- 0.1% in 1999. Decreased daily smoking was observed in all demographic subgroups except young adults. Among college graduates, the daily smoking prevalence in 1999 was 6.4 +/- 0.4%, a level previously observed only among U.S. physicians. In 1999, nearly 30% of current smokers did not smoke daily, and more than 60% said they now smoked less than previously. In 1999, self-reported cigarette consumption was inversely related to believing SHS is harmful to nonsmokers, having a smoke-free workplace, and living in a smoke-free home. In California, tobacco control strategies that educated the population about SHS and resulted in smoking restrictions may have led continuing smokers to smoke less, which should reduce the harm from smoking to the public health in the long term. PMID- 12583357 TI - Computed tomography for coronary calcification: mammogram of the heart or does the cup runneth over? PMID- 12583358 TI - International Conference on Harmonisation; revised guidance on Q3A impurities in new drug substances; availability. Notice. AB - The Food and Drug Administration (FDA) is announcing the availability of a revised guidance entitled "Q3A(R) Impurities in New Drug Substances." The revised guidance, which updates a guidance on the same topic published in the Federal Register of January 4, 1996 (the 1996 guidance), was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The revised guidance clarifies the 1996 guidance, adds information, and provides consistency with more recently published ICH guidances. The revised guidance is intended to provide guidance to applicants for drug marketing registration on the content and qualification of impurities in new drug substances produced by chemical syntheses and not previously registered in a country, region, or member State. PMID- 12583359 TI - Bush rule makes fetuses eligible for health benefits. PMID- 12583361 TI - Myocardial perfusion imaging in Denmark: activity from 1997 to 2001 and current practice. AB - A questionnaire was sent to all departments of nuclear medicine in Denmark (n=20) asking for details of myocardial perfusion imaging (MPI), including the number of patients examined each year from 1997 to 2001 and the current clinical and technical practice. All (100%) departments replied, and the survey thus covers all MPI performed in Denmark during the period in question. The number of MPI studies (examined patients) was 2,531 in 1997 (0.47 MPI/1,000/year) and 4,961 (0.93 MPI/1,000/year) in 2001, which is a doubling in activity in 5 years. Nineteen (95%) of the Danish departments performed MPI in 2001, and 14 (74%) of these reported that activity had increased over the past 5 years. MPI activity was unevenly distributed between hospitals and regions. In 2001, the university hospitals in the central Copenhagen region (capital) accounted for the highest MPI activity (2.00/1,000/year), while the non-university hospitals in general had the lowest activity rate (0.73/1,000/year). The most pronounced increment found in the period was observed in the university hospitals outside Copenhagen, where activity increased by 300% from 0.44/1,000/year in 1997 to 1.33/1,000/year in 2001. All departments providing MPI used tomographic acquisition technique and all departments used technetium tracers. The more sophisticated techniques of MPI - gated acquisition, attenuation correction and iterative reconstruction - were used in 74%, 32% and 42% of departments, respectively. The stress mode in perfusion studies was dipyridamole/adenosine in 76%, exercise in 18% and dobutamine in 6%. Despite these encouraging figures, MPI activity for 2001 remained well below what is recommended by other national and international societies. The anticipated further increase in nuclear cardiology is encouraging, but the nuclear medicine community needs to address the issues that prevent it from keeping up with demand. In general, the restricted camera time and the limited number of trained personnel explain the excessive waiting lists in Denmark. PMID- 12583360 TI - Compliance with patients' end-of-life wishes by nursing homes in New York City with conscience policies. AB - Nursing home patients have a constitutional right to refuse treatment. The Patient Self-Determination Act confirmed that right. State laws address the obligations of health care providers and facilities to honor that right. The New York State law is more specific than those of many other states. It allows exemptions for "reasons of conscience" and imposes a number of requirements on nursing homes claiming such an exemption, including the transfer of a patient to a home that will honor an end-of-life wish. This study, conducted by FRIA, investigated the refusal of some nursing homes in New York City to carry out patients' end-of-life wishes because of conscience-based objections. The study also investigated the willingness of homes which did not have such policies to accept patients transferring from a home with a policy so that the patient's end of-life wishes would be honored. Implications for administrators, policy makers, and regulators are discussed. PMID- 12583362 TI - Should high hTg levels in the absence of iodine uptake be treated? PMID- 12583363 TI - Bibliography. Current world literature. Nutrition and metabolism. PMID- 12583364 TI - Hormone replacement therapy not a significant breast cancer risk? Concerns regarding a Society of Obstetricians and Gynaecologists of Canada press release. PMID- 12583365 TI - Infection is not specific for bacterial osteo-articular infective pathology. PMID- 12583366 TI - Sentinel lymph node detection in cases of vulvar malignancy. PMID- 12583367 TI - European Society of Intensive Care Medicine statement: intensive care medicine in Europe--structure, organisation and training guidelines of the Multidisciplinary Joint Committee of Intensive Care Medicine (MJCICM) of the European Union of Medical Specialists (UEMS). AB - This article describes the structures and institutions in the European Union by which professional training and qualification in medical specialities will be harmonised. All main medical specialities are represented in the European Union of Medical Specialists (UEMS) by speciality sections. For intensive care medicine, as a multidisciplinary speciality, a new structure of a Multidisciplinary Joint Committee of Intensive Care Medicine (MJCICM) within the UEMS was established in 1999. In this MJCICM the European Society of Intensive Care Medicine and the European Society of Paediatric and Neonatal Intensive Care are represented by delegates without voting capacity in a Standing Advisory Board. Statements and recommendations which the MJCICM has worked out until now are presented: Definitions of intensive care medicine, structural conditions for education and training, continuing medical education, criteria for accreditation of intensive care medicine training centres, common core curriculum for optional specialist training in intensive care medicine, as well as an intensive care units accreditation visiting programme and standards for medical treatment and nursing care. PMID- 12583368 TI - Statement of the 4th International Consensus Conference in Critical Care on ICU Acquired Pneumonia--Chicago, Illinois, May 2002. PMID- 12583369 TI - The Soviet-Russian space suits a historical overview of the 1960's. AB - The development of protective suits for space use started with the Vostok-suit SK 1, first used by Yu. Gagarin on April 12, 1961, and then used on all subsequent Vostok-flights. The technical background for the design of these suits was the work on full pressure protective suits for military pilots and stratospheric flights in the 1930's through 50's. The Soviet-Russian space programme contains a large number of 'firsts', and one of the most well known is the first EVA by Leonov in 1965. This event is also the starting point for a long series of space suit development for Extravehicular Activities over the last 35 years. The next step to come was the transfer in void space of crew members between the two spacecraft Soyuz 4 and 5 in 1969. As has later become known this was an essential element in the planned Soviet lunar exploration programme, which in itself required a new space suit. After the termination of the lunar programme in 1972, the space suit development concentrated on suits applicable to zero-gravity work around the manned space stations Salyut 6, Salyut 7 and MIR. These suits have become known as the ORLAN-family of suits, and an advanced version of this suit (ORLAN-M) will be used on the International Space Station together with the American EMU. This paper covers the space suit development in the Soviet Union in the 1960's and the experience used from the pre-space era. PMID- 12583370 TI - Endocrine related resources from the National Institutes of Health. PMID- 12583371 TI - The presence of shock defines the threshold to initiate thrombolytic therapy in patients with pulmonary embolism. PMID- 12583372 TI - National Hormone and Peptide Program. National Institute of Diabetes and Digestive and Kidney Diseases. Peptides hormones, subunits, antisera and reagents available. PMID- 12583373 TI - The case for thrombolysis in acute major pulmonary embolism: hemodynamic benefits and beyond. PMID- 12583374 TI - Intrinsic (or auto-) positive end-expiratory pressure during spontaneous or assisted ventilation. PMID- 12583375 TI - Clearance of vancomycin during high-volume haemofiltration: impact of pre dilution. AB - OBJECTIVE: To measure the sieving coefficient (SC) and clearance of vancomycin during high-volume haemofiltration (HVHF) and to evaluate the impact of different pre-dilution regimens on these variables. DESIGN AND SETTING: Prospective interventional study in the intensive care unit in a tertiary university hospital. PATIENTS: Seven patients with septic shock and multi-organ dysfunction. INTERVENTIONS: HVHF (6 l/h fluid exchange) was performed in septic shock patients using variable proportions of their replacement fluid in pre- and post-dilution mode. MEASUREMENTS AND RESULTS: Pre-filter, post-filter and ultrafiltrate vancomycin concentrations were measured simultaneously, and SC and clearance calculated. The measurements were repeated following each change in the proportion of pre-dilution fluid. SC steadily decreased as the proportion of pre dilution decreased, changing from 0.76 in pure pre-dilution to 0.57 in pure post dilution (p=0.0004). Clearance, however, increased with decreasing pre-dilution fluid rate, from 53.9 ml/min at pure pre-dilution to 67.2 ml/min at 2 l/h pre dilution with 4 l/h post-dilution. CONCLUSIONS: HVHF achieves high vancomycin clearances, which despite some deterioration in SC increase with the proportion of replacement fluid given post-filter. Clinicians applying HVHF need to be aware of such clearances to avoid inadequate vancomycin dosing and to adjust therapy according to variations in HVHF technique. PMID- 12583376 TI - Optimal lung volume strategy: comment on "High-frequency oscillatory ventilation in experimental lung injury: effects on gas exchange" by Dembinski et al. PMID- 12583378 TI - Comments on: A new prognostic scoring system for meningococcal septic shock in children. PMID- 12583379 TI - Sonographic appearance of ARDS. Letter: ref. Tsubo T, Yatsu Y, Suzuki A, Iwakawa T, Okawa H, Ishihara H, Matsuki A (2001). Daily changes of the area of density in the dependent lung region--evaluation using transesophageal echocardiography. Intensive Care Med 27:1881-1886. PMID- 12583380 TI - Sample introduction mode: another source of error in saline PCO2 measurement. PMID- 12583381 TI - Towards the VI European Community Framework Programme and the European Research Area. PMID- 12583382 TI - Tacrolimus-induced life-threatening arrhythmia in a pediatric liver-transplant patient. PMID- 12583383 TI - Early-onset Pneumocystis carinii pneumonia complicating Norwood II operation in a 3-month-old patient. PMID- 12583384 TI - Fatal catecholamine myocarditis in a child with severe scalding injury. PMID- 12583385 TI - Long-term operation of "Orlan" space suits in the "Mir" orbiting station: experience obtained and its application. AB - The started assembly of the International Space Station (ISS) and its further operation will call for a great number of extravehicular activity sorties (EVA) to be performed by ISS crews. Therefore, of great importance is to make use of the EVA experience gained by cosmonauts in the process of 15-year operation of the Mir orbiting station (OS). Over the 15-year period, Mir crewmembers wearing Orlan type semi-rigid space suits have accumulated 158 man/sorties from the orbiting station. Crewmembers used 15 suits in orbit and some of the suits were in operation for more than 3 years. The paper presents principal design features, which provide effective and safe operation of orbit-based suits, and briefly describes procedures for preparation and maintenance of suit systems, which ensure long-term operation of space suit in orbit. The paper gives results of the space suit modifications, presents suit performance characteristics and lists novel or upgraded components of the space suit and its systems. The paper also summarizes improvements in the Orlan type suits described in some earlier publications. They refer, in the first run, to the improvement of space suit operations characteristics and reliability, and the utilization of the Orlan type space suit in the ISS program. The paper analyses the experience gained and drawbacks detected and observations made, and gives statistical data on long-term space suit operations aboard the Mir station. The paper reviews certain problems in the process of EVAs performed from the station, and describes the ways of their solution as applied to the further utilization of the suit within the ISS program. PMID- 12583386 TI - Adaptive environmental control for optimal results during plant microgravity experiments. AB - The SVET Space Greenhouse (SG)--the first and the only automated plant growth facility onboard the MIR Space Station in the period 1990-2000 was developed on a Russian-Bulgarian Project in the 80s. The aim was to study plant growth under microgravity in order to include plants as a link of future Biological Life Support Systems for the long-term manned space missions. An American developed Gas Exchange Measurement System (GEMS) was added to the existing SVET SG equipment in 1995 to monitor more environmental and physiological parameters. A lot of long-duration plant flight experiments were carried out in the SVET+GEMS. They led to significant results in the Fundamental Gravitational Biology field- second-generation wheat seeds were produced in the conditions of microgravity. The new International Space Station (ISS) will provide a perfect opportunity for conducting full life cycle plant experiments in microgravity, including measurement of more vital plant parameters, during the next 15-20 years. Nowadays plant growth facilities for scientific research based on the SVET SG functional principles are developed for the ISS by different countries (Russia, USA, Italy, Japan, etc.). A new Concept for an advanced SVET-3 Space Greenhouse for the ISS, based on the Bulgarian experience and "know-how" is described. The absolute and differential plant chamber air parameters and some plant physiological parameters are measured and processed in real time. Using the transpiration and photosynthesis measurement data the Control Unit evaluates the plant status and performs adaptive environmental control in order to provide the most favorable conditions for plant growth at every stage of plant development in experiments. A conceptual block-diagram of the SVET-3 SG is presented. PMID- 12583387 TI - Using the PROGRESS transport spacecraft in structure of the International Space Station for realization of scientific experiments under microgravity conditions. AB - The problem is considered of using the PROGRESS transport spacecraft, which will deliver the payload on the ISS, as a free flying platform for realization of space experiments. For maintenance of the ISS 5-6 PROGRESS flights per year are planned. Usually after delivery of the payload the PROGRESS undocks from the ISS and burns down in the Earth atmosphere. However, the operating conditions of its onboard systems allow to prolong operation and to make free flight near to the station and repeatedly to be docked to it. It is offered to use this possibility for performing experiments on Material Science. PMID- 12583388 TI - Space technologies in routine telemedicine practice: commercial approach. PMID- 12583389 TI - Following the water, the new program for Mars exploration. AB - In the wake of the loss of Mars Climate Orbiter and Mars Polar Lander in late 1999, NASA embarked on a major review of the failures and subsequently restructured all aspects of what was then called the Mars Surveyor Program--now renamed the Mars Exploration Program. This paper presents the process and results of this reexamination and defines a new approach which we have called "Program System Engineering". Emphasis is given to the scientific, technological, and programmatic strategies that were used to shape the new Program. A scientific approach known as "follow the water" is described, as is an exploration strategy we have called "seek--in situ--sample". An overview of the mission queue from continuing Mars Global Surveyor through a possible Mars Sample Return Mission launch in 2011 is provided. In addition, key proposed international collaborations, especially those between NASA, CNES and ASI are outlined, as is an approach for a robust telecommunications infrastructure. PMID- 12583391 TI - One hundred US EVAs: a perspective on spacewalks. AB - In the 36 years between June 1965 and February 2001, the US human space flight program has conducted 100 spacewalks, or extravehicular activities (EVAs), as NASA officially calls them. EVA occurs when astronauts wearing spacesuits travel outside their protective spacecraft to perform tasks in the space vacuum environment. US EVA started with pioneering feasibility tests during the Gemini Program. The Apollo Program required sending astronauts to the moon and performing EVA to explore the lunar surface. EVA supported scientific mission objectives of the Skylab program, but may be best remembered for repairing launch damage to the vehicle and thus saving the program. EVA capability on Shuttle was initially planned to be a kit that could be flown at will, and was primarily intended for coping with vehicle return emergencies. The Skylab emergency and the pivotal role of EVA in salvaging that program quickly promoted Shuttle EVA to an essential element for achieving mission objectives, including retrieving satellites and developing techniques to assemble and maintain the International Space Station (ISS). Now, EVA is supporting assembly of ISS. This paper highlights development of US EVA capability within the context of the overarching mission objectives of the US human space flight program. PMID- 12583392 TI - Canada and the International Space Station program: overview and status. AB - The twelve months since IAF 2000 have been perhaps the most exciting, challenging and rewarding months for Canada since the beginning of our participation in the International Space Station program in 1984. The highlight was the successful launch, on-orbit check out, and the first operational use of Canadarm2, the Space Station Remote Manipulator System, between April and July 2001. The anomalies encountered and the solutions found to achieve this success are described in the paper. The paper describes, also, the substantial progress that has been made, during the twelve months since IAF 2000, by Canada as it continues to complete work on all flight-elements of its contribution to the International Space Station and as we transition into real-time Space Station operations support and Canadian utilization. Canada's contribution to the International Space Station is the Mobile Servicing System (MSS), the external robotic system that is key to the successful assembly of the Space Station, the maintenance of its external systems, astronaut EVA support, and the servicing of external science payloads. The MSS ground segment that supports MSS operations, training, sustaining engineering, and logistics activities is reaching maturity. The MSS Engineering Support Center and the MSS Sustaining Engineering Facility are providing real time support for on-orbit operations, and a Canadian Payloads Telescience Operations Center is now in place. Mission Controllers, astronauts and cosmonauts from all Space Station Partners continue to receive training at the Canadian Space Agency. The Remote Multi Purpose Room, one element of the MSS Operations Complex, will be ready to assume backroom support in 2002. Canada has completed work on identifying its Space Station utilization activities for the period 2000 through 2004. Also during the past twelve months the CSA drafted and is proceeding with the approval of a Canadian Space Station Commercialization Policy. Canadian astronauts have now participated in three ISS assembly missions- Julie Payette on STS-96, Marc Garneau on STS-97, and Chris Hadfield on STS-100 in April 2001 during which he performed Canada's first EVA and the successful installation of the Space Station Remote Manipulator System. PMID- 12583393 TI - [The 25th annual scientific meeting of the Japanese Society of Hypertension. Tokyo, Japan. October 11-13, 2002. Program and Abstracts]. PMID- 12583394 TI - CASH 2021: commercial access and space habitation. AB - Issues about commercialization of space have been a growing concern in the past decade for the space community. This paper focuses on the work from a team of 51 students attending the Summer Session Program of the International Space University in Bremen, Germany. CASH 2021 (Commercial Access and Space Habitation) documents a plan that identifies commercial opportunities for space utilization that will extend human presence in space, and will chart the way forward for the next 20 years. The group selected four commercial sectors that show the most promise for the future: tourism, entertainment, space system service, assembly and debris removal, and research and development/production. The content of this document presents the results of their research. Historical activities in each of the commercial sectors are reviewed along with the current market situation. To provide a coherent background for future commercialization possibilities a scenario has been developed. This scenario includes a postulated upon ideal future and includes social, political and economic factors that may affect the space industry over the timeline of the study. The study also presents a roadmap, within the limited optimistic scenario developed, for the successful commercialization of space leading to future human presence in space. A broad range of commercially viable opportunities, not only within the current limits of the International Space Station, but also among the many new developments that are expected by 2021 are discussed. PMID- 12583395 TI - Halting the growth in diagnostic testing. PMID- 12583396 TI - Air pollutants, exercise, and risk of developing asthma in children. PMID- 12583397 TI - An interventional program for diagnostic testing in the emergency department. PMID- 12583398 TI - Evidence-based practice...what is it and how do I do it? PMID- 12583399 TI - Germination and growth of lettuce (Lactuca sativa) at low atmospheric pressure. AB - The response of lettuce (Lactuca sativa L. cv. Waldmann's Green) to low atmospheric pressure was examined during the initial 5 days of germination and emergence, and also during subsequent growth to vegetative maturity at 30 days. Growth took place inside a 66-l-volume low pressure chamber maintained at 70 kPa, and plant response was compared to that of plants in a second, matching chamber that was at ambient pressure (approximately 101 kPa) as a control. In other experiments, to determine short-term effects of low pressure transients, plants were grown at ambient pressure until maturity and then subjected to alternating periods of 24 h of low and ambient atmospheric pressures. In all treatments the partial pressure of O2 was maintained at 21 kPa (approximately the partial pressure in air at normal pressure), and the partial pressure of CO2 was in the range 66.5-73.5 Pa (about twice that in normal air) in both chambers, with the addition of CO2 during the light phase. With continuous exposure to low pressure, shoot and root growth was at least as rapid as at ambient pressure, with an overall trend towards slightly greater performance at the lower pressure. Dark respiration rates were greater at low pressure. Transient periods at low pressure decreased transpiration and increased dark respiration but only during the period of exposure to low pressure. We conclude that long-term or short-term exposure to subambient pressure (70 kPa) was without detectable detriment to vegetative growth and development. PMID- 12583400 TI - Effects of space food bar feeding on bone mass and metabolism in normal and unloaded rats. AB - During spaceflights in the shuttle, rats are provided specific food bars. To determine whether this diet allows normal body and skeletal growth, we used four groups of rats fed either standard pellet food or space food bars during a 2-wk unloading experiment. We recorded food intake, body weight, tibial bone mass, and mineral content by ash analyses, cancellous bone volume, and cell activities by histomorphometry. We found that food intake was not different when comparing the two types of food, but that suspended animals had a lower food intake than normal loaded animals. Body weight and bone mass were found lower in suspended animals than in normal loaded animals. Finally, longitudinal growth rate, cancellous bone volume, and bone formation rate were lower in suspended animals, irrespective of the type of food. These results show that space food bar feeding did not affect normal body and skeletal growth, and that body and bone changes due to unloading were not significantly different in animals fed space food bars and standard food. PMID- 12583401 TI - Tribute to Dr. Carl T. Brighton. PMID- 12583402 TI - Human motion planning based on recursive dynamics and optimal control techniques. AB - This paper presents an efficient optimal control and recursive dynamics-based computer animation system for simulating and controlling the motion of articulated figures. A quasi-Newton nonlinear programming technique (super-linear convergence) is implemented to solve minimum torque-based human motion-planning problems. The explicit analytical gradients needed in the dynamics are derived using a matrix exponential formulation and Lie algebra. Cubic spline functions are used to make the search space for an optimal solution finite. Based on our formulations, our method is well conditioned and robust, in addition to being computationally efficient. To better illustrate the efficiency of our method, we present results of natural looking and physically correct human motions for a variety of human motion tasks involving open and closed loop kinematic chains. PMID- 12583403 TI - [Reopening the 'black box': discontinuity and continuity in the discourse on AIDS in the USA (1987-98)]. AB - The article has the specific goal of examining AIDS Coalition to Unleash Power's ideas and actions. In a broader scope, it tries to discuss issues related to scientific theory and practice concerning AIDS in the United States. The sources used for the present study consist of printed and electronic documents produced by AIDS Coalition to Unleash Power (ACT UP/NY). After a period of great radicalization, ACT UP developed a less of confronting position. Among other factors, this was brought about by the changes that took place in the North American political scenario in the beginning of the 1990's, as well as by internal dissent. The analyzed data allow us to say that the group has been quite successful in relation to the production and the distribution of medicines for the treatment of AIDS. They also indicate partial changes in the competitive game for the monopoly of 'truths' about the disease. PMID- 12583404 TI - Should high hTg levels in the absence of iodine uptake be treated? PMID- 12583405 TI - Passionate about patients. PMID- 12583406 TI - [Does a reduced kidney function (creatinine clearance 10-15 ml/min), without acidosis, increase the risk for the rise of lactic acidosis caused by metformin?]. PMID- 12583407 TI - Healthy premiums. PMID- 12583408 TI - [Does the vitamin B12 deficiency caused by metformin disappear again after stopping this drug?]. PMID- 12583409 TI - Treatment of critical bleeding in the future intensive care unit. PMID- 12583410 TI - [Rockefeller Foundation and health assistance in Sao Paulo (1920-30): historical perspectives]. AB - The present paper aims at presenting a short history of health policies implemented in Sao Paulo in the 1920's and 1930's, trying to reach a more accurate understanding of the processes that led to the formulation of governmental health policies. The paper emphasizes Geraldo Horacio de Paula Sousa's contribution to the history of health administration in Sao Paulo in the first half of the twentieth century and his proposition of a new model of health assistance based on health units that would work as the structural axis of all public health activities in the state. Finally, it focuses on the support given by Rockefeller Foundation to Paula Sousa's activities and on the disputes involved in the establishment of public health policies. PMID- 12583411 TI - Lipoprotein (a) and coronary artery disease in Indians. PMID- 12583412 TI - Diagnostic stromal histomorphology in fibroepithelial breast lesions: a fresh perspective. AB - Histopathologic parameters were studied in fibroepithelial breast lesions comprising of Fibroadenomas (FAs), Juvenile cellular fibroadenomas (JCFs) as well as benign, borderline and malignant phyllodes tumours (B/Bo/MPTs). H & E stained sections of 67 cases (25 ICFA, 5 Mixed FA, 10 PCFA, 12 JCF and 15 PT) were retrieved from archival material and the microscopic features of each lesion, reviewed by two independent observers. A diagnosis was arrived at by consensus. Diagnostic criteria were laid down for each of the above lesions based on a statistical analysis of data obtained from a previously drawn up semiquantitative grading system for various stromal histomorphologic parameters. An increasing grade of stromal cellularity, pleomorphism, overgrowth and mitotic index was observed, from the benign to the borderline and overtly malignant lesions included in the spectrum. A semiquantitative grading of relevant stromal histomorphologic variables was found to be of immense diagnostic significance in fibroepithelial breast lesions and served to elucidate cases falling into the diagnostic grey zones of the spectrum. PMID- 12583413 TI - Immune complex mediated lesions in experimental Kala azar: an ultrastructural study. AB - Immune complexes play an important role in causation of renal lesions in various diseases. Circulating immune comlexes (CIC) are described in Kala azar. Role of CIC in pathogenesis of Kalaazar is discussed in present study. BALB/C mice were experimentally infected with L. donovani promastigotes. After visceralisation of infection, sera and kidneys of infected mice were preserved. Leishmanial antigen specific CIC could be demonstrated in 100% of infected mice by PEG ELISA, while they were absent in control mice. Ultrastructural pattern of renal lesions in infected mice showed presence of focal small electron dense deposits in glomerular basement membrane and subepithelial space, resembling immune complexes (humps). Rarely subendothelial and mesangial hypercellularity was present. These findings point towards a definite role of CIC in pathogenesis of renal lesions in Kalaazar. PMID- 12583414 TI - Salmonella 3, 10:r: toxicity in rabbit ileum and liver by light and electron microscopy. AB - Salmonella 3, 10:r:- (a monophasic variety of otherwise diphasic serotypes such as S. weltevreden and S. simi) Cell_free filtrate, when introduded into rabbit ligated ileal loops causes fluid exsorption, as studied 18-hr after treatment. Light microscopic histology of treated ileum shows denudation of the columnar epithelium at several places, thereby allowing the passage of the toxic principle into circulation. An important target organ, liver shows extensive centrilobular necrosis, as observed by light microscopy. Transmission electron microscopy of ileum reveals opening of membrane junctions between the adjacent cells of epithelial lining of the treated ileum at places, and focal devitalization including formation of intra-cellular membranous inclusinos. Electron microscopy of liver shows extensive damage and swelling of cytoplasmic membranes. However, the areas of darkly staining lamellae of granulated endoplasmic reticulum are also seen in stacks as will as dispersed. These studies stress that Salmonella toxic substances can cause extensive damage to intestine and liver both. PMID- 12583415 TI - Accuracy of cytological findings in abnormal cervical smears by cytohistologic comparison. AB - To investigate the accuracy rates of cytology in abnormal cervical smears and the factors contributing to a discrepant diagnosis between cytology and histology repots of cervical intraepithelial and invasive neoplasm. During the four-year period 1993 to 1996, abnormal cervical smear findings, which were followed by cervical biopsy, were available in 709 patients. The cytology and histology slides were reviewed in each case. The accuracy rates of cytology before and after review were investigated. The accuracy rate of cytology was 48%. Following review it became 56%, mainly due to a reduction in the number of cases in which the smear showed a lesser degree of CIN than did the biopsy. The proportion of cases in which the cytological impression of CIN was more severe than the histology was minimally altered. The results suggest that difficulty in the interpretation of cervical smear as well as sampling errors are responsible for reduced accuracy even in smears which are considered representative of the pathological process. PMID- 12583416 TI - Salmonella types and antibiotic susceptibility: a six months survey. AB - A total of 51 Salmonella strains were isolated during the six month period of May 1998 till October 1998. Of these, 41 (80.3%) were Salmonella typhi, 5 (9.8%) Salmonella pararyphi A,2 (3.9%) Salmonella worthington and 1 (1.9%) Salmonella senftenberg. The prevalent phage and biotype of Salmonella typhi was E1 (75%) and type 1 (90.2%) respectively. The commonest pattern of multiple drug resistance in Salmonella typhi was ACCoT and 92.5% of these belonged to phage E1. Out of the five Salmonella paratyphi A, one belonged to phage type 1 and the others were untypable. Similarly both the strains of Salmonella typhimurium were untypable. Thus the predominant isolate was Salmonella typhi and the commonest phage and biotype were E1 and biotype1 respectively. PMID- 12583417 TI - Immunoglobulin-M estimation and C-reactive protein detection in neonatal septicemia. AB - Immunoglobulin-M estimation, and C-Reactive Protein test and blood culture were performed on 57 neonates, clinically suspected to have septicemia. IgM level of > or = 20 mg/dl was found in 58.62% cases and CRP test was positive in 68.98% of culture proven sepsis. Blood culture was positive in 29 cases, with the predominant organism being Klebsiella. Of the 2 tests, CRP had a higher sensitivity (68.97%) while IgM estimation had a higher specificity (82.14%). When the 2 tests were considered together the sensitivity and specificity further increased. Death rate among neonates with IgM levels < 20 mg/dl was 5 times higher than those with elevated IgM levels. PMID- 12583418 TI - Study of coagulase negative staphylococci isolated from blood and CSF cultures. AB - Coagulase negative Staphylococci (CONS) which were considered as laboratory contaminants and normal flora of skin in man, have emerged as opportunistic pathogens. The infection with CONS has been reported since 1950 with increasing frequency and has been implicated as the causative agents of certain categories of patients viz, neonates with sepsis, cardiac patients with prosthetic valves, immunocompromided patients which include end-renal stage disease, and renal transplantation, burns and cancer patient. These are causing problems to clinicians because of their drug resistance. 180 strains of CONS isolated from blood and CSF during the period of 2 years (Jan 1997-Dec 1998) were studied. Not only they were resistant to Penicillin (P), Ampicillin, (Amp), Oxacillin (Ox), but also developing resistance to Vancomycin (Van) which pose a therapeutic problem. So this study was undertaken and this area needs further exploration. PMID- 12583419 TI - Bone marrow necrosis in paediatric patients. AB - Necrosis of bone marrow is a very rare condition with a prevalence ranging from 0.15 to 7 percent. It is mostly not compatible with life. We diagnosed four cases of necrotic marrow out of 830 bone marrow aspirations done in last five years of time. All were paediatric patients having peripheral pancytopenia. Three out of four patients expired within one month of diagnosis. PMID- 12583420 TI - The mechanism of ampicillin resistance in enterobacteriaceae isolated in Vellore. AB - Ampicillin resistance in clinical isolates of enterobacteriaceae has been on the increase in the last decade. A study was carried out to delineate the possible mechanisms of this resistance. The method employed was isoelectric focussing to observe the electrophoretic mobility of the enzyme responsible and this showed that TEM 1 betalactamase was the cause irrespective of the species. PMID- 12583421 TI - Platelet transfusion practice during dengue fever epidemic. AB - Blood components especially platelet concentrates due to their short shelf life are frequently in limited supply. Appropriate use of blood components is required to ensure their availability for needy patients as well as to avoid the unnecessary risk of transfusion-transmitted diseases. Medical audit of blood transfusion practice, which forms an important part of quality assurance programme in a transfusion centre, can provide grounds for improvement in transfusion medicine practice. During the epidemic of dengue fever in Oct., 1996, 1837 patients were admitted as dengue haemorrhagic fever in a teaching hospital in Delhi. Two hundred and eight patients (11.3%) were given platelet transfusions. Retrospective analysis of these platelet transfusions was done. It was observed that in only 52 (25%) out of 208 patients the information on platelet counts was provided. History of active bleeding was obtained only in 65 (31.2%) patients. About 35% patients received unnecessary prophylactic transfusions and during 89% of the transfusion episodes inappropriate dose of platelet concentrate was given. Information regarding post-transfusion recovery could be obtained in only 16.5% of transfusion episodes. The study emphasises the need for development of specific guidelines for transfusion of blood components, constant interaction and co-ordination amongst clinicians and transfusion centre for implementation of these guidelines, and a regular medical audit to review the optimal utilisation of blood components. PMID- 12583422 TI - P 53 and PCNA in Non Hodgkin's lymphoma--an immunohistochemical evaluation. AB - P 53 positivity and PCNA labelling index were studied in 50 cases of node based Non Hodgkin's lymphomas, P 53 positivity was observed in a spectrum of these disorders, suggesting that P 53 mutations play a role in the genesis of these tumours. P 53 positivity incresed from low through intermediate to high grade tumours and thus may be of prognostic value in these lesions. PCNA labelling index (LI) was higher in high grade tumours. P 53 and PCNA immunoreactivity showed a relationship in that PCNA LI was seen to be more than 30% in P 53 positive tumours. P 53 positivity appears to be related to the subtype and proliferation rate of NHL. PMID- 12583423 TI - A study of yeast carriage on hands of hospital personnel. AB - The present study was conducted by culture with a modified broth wash technique to examine the frequency of yeast carriage on the hands of 60 nurses and 35 nonnursing hospital employees. Seventy two percent of the nurses and 80% of the nonnurses were harbouring yeast on their hands. Candida spp. were isolated in 57% on of nurses and 34% of nonnurses. Ninety percent of nurses working in nursing home care unit (NHCU), 50% working in intensive care unit (ICU) and 75% working in outpatient department (OPD) carried yeasts on their hands. Only 80% of nonnurses staff harboured yeasts on their hands. PMID- 12583424 TI - Fasciolopsiasis--a persisting problem in eastern U.P.--a case report. AB - Fasciolopsiasis, or infection by the intestinal fluke, Fascilopsis buski, is endemic in the eastern states of our country. While it is by no means a rarity, especially in the rural set up, awareness regarding this common parasitic infestation is still a much-needed entity. The importance of a strong degree of suspicion and early diagnosis cannot be over emphasised, if a successful campaign is to be launched in its control. With this as our central theme, we proceed to report a case of and unsually heavy Fasciolopsis buski infection in our hospital, which had failed to be diagnosed in a semi urban setup in UP. PMID- 12583425 TI - Fasciolopslasis--a re-emerging infection in Azamgarh (Uttar Pradesh). AB - Fasciolopsiasis is endemic in the far east. In India, there have been a few reports of the infection, prior to the 1990's. We report two cases from Azamgarh district of Uttar Pradesh. Both the cases were from nearby villages where water chestnuts are cultivated. These may be a source of infection. Pigs are commonly observed in these areas and and may be the source of ova. The only missing link is the finding of infected snails. Presence of at least three cases (one reported earlier) in the area indicates the potential for the infection to re-emerge. Further epidemiological studies are needed to analyse the various ecological factors of transmission. Fasciolopsiasis is endemic in China, Taiwan, Vietnam and Thailand. In India, (Fascilopsis buski) infections in man have been reported earlier from Assam, Maharashtra, Tamil Nadu and parts of Uttar Pradesh. However, to the best of our knowledge, no such reports have been made since 1990's. We herewith report two recent cases from district Azamgarh, Uttar Pradesh (U.P.), India. Factors, such as cultivation of water chestnuts, presence of snails as intermediate hosts and pigs as definitive host in this geographical area seem to be suggestive of an endemic focus and thus needs further epidemiological survey for preventive and control measures, at the earliest. PMID- 12583426 TI - Human ocular dirofilariasis in Kerala Southern India. AB - A twenty-six year old female presented to the Ophthalmology OPD with swelling of the upper eyelid. On excision, two long, thin, dead worms were removed. Microscopic and HPE showed the worms to belong to the Dirofilaria species. PMID- 12583427 TI - Cerebral chromoblastomicosis--a rare case report of cerebral abscess and brief review of literature--a case report. AB - A rare case of Cerebral Chromomycosis caused by chromogenic fungus Cladosporium trichoides in a 35 year old male with classical presentation of cerebral abscess is being presented. The case report lays emphasis on the histological diagnosis of chromogenic fungus in the wall of the abscess cavity, surgically removed from a well delineated circumscribed lesion in the frontal lobe of the cerebrum. The causative fungus could be detected even in unstained paraffin sections. The diagnosis could be made only after surgical removal and histopathological examination. The mycological culture could not be made as the material was received in formaldehyde fixative. The unique features of the case is its recurrence free uneventful survival five years after surgical excision. This is probably the fifth reported case of cerebral chromomycosis from India and first of its type from arid zone of Rajasthan. PMID- 12583428 TI - Carcinoma arising within fibroadenoma--a case report. AB - A rare case of ductal carcinoma-in-situ in multiple fibroadenomas of the breast in a 43 year old female is reported. PMID- 12583429 TI - Wolman's disease--a case report. AB - Wolman's disease is a rare autosomal recessive lysosomal storage disorder. We report a case, which we identified with foamy histiocytes in bone marrow and adrenal calcification in radiological imaging. The diagnosis can be made on minimal investigation when clinically suspected. But cytogenetic study is required to substantiate the diagnosis further. PMID- 12583430 TI - Fibrous histiocytoma of orbit: a case report. AB - A fifty year male (K.S.) presented with protrusion (Proptosis) of left eye which was gradually increasing in size for the last one year. The swelling was accompanied by pain for the last fifteen days. Preoperative X-ray & computed tomography showed erosion of frontal sinus & roof of maxillary sinus. Clinical diagnosis of a malignant tumour of eye was made & enucleation of the left eye, along with the mass was done. The specimen was received in the pathology department on 16.9.98. Grossly, encapsulated mass attached with the intact eye ball was received. The mass measured 5x3x2.5 cms. Microscopic examination revealed a benign spindle cell neoplasm and a diagnosis of fibrous histiocytoma (dermatofibroma) was made. PMID- 12583431 TI - Dermoid cyst of the testis--a rare entity. PMID- 12583432 TI - Minimally invasive hyalinising trabecular carcinoma. PMID- 12583433 TI - Cholera epidemic in and around Loni a rural area in western Maharashtra. PMID- 12583434 TI - Molecular pathogenesis and prognostic factors in endometrial carcinoma. AB - Endometrial carcinoma is today among the most common gynecologic malignancies in industrialized countries. In order to improve the treatment and follow-up of these patients, various prognostic factors have been extensively studied. Patient age, stage of disease, histologic type and histologic grade have been shown to influence survival significantly, and the prognostic impact of these traditional clinicopathologic variables is well established. In addition, parity, hormone receptor concentration in the tumor, DNA ploidy and morphometric nuclear grade have all been found to influence prognosis. Information about DNA ploidy has especially been used in the clinical situation to determine individualized treatment. The prognostic significance of markers for tumor cell proliferation, cell cycle regulation (p53, p21 and p16) and angiogenesis is discussed as well as the molecular basis of endometrial carcinoma. In conclusion, several prognostic markers have been identified. It is likely that the information derived from these tumor biomarkers will reduce the need for extensive surgical staging and adjuvant treatment in endometrial carcinoma. PMID- 12583435 TI - Calibration of fusidic acid disk diffusion susceptibility testing of Staphylococcus areus. AB - Single strain regression analysis, SRA, was used to calibrate disk diffusion fusidic acid susceptibility testing of Staphylococcus aureus in two laboratories using different standard methods but the same interpretative MIC limits. SRA equation constants were calculated using five different fusidic acid disk contents (1.5, 5, 15, 50, 150 microg). These disks were tested on five separate occasions against quality control strain S. aureus ATCC 29213. The National Committee for Clinical Laboratory Standards (NCCLS) method was employed in Tartu, Estonia (TE) and the Swedish Reference Group for Antibiotics (SRGA) method in Sweden at the Karolinska Hospital (KS). SRA constants obtained were used for calculating zone breakpoints corresponding to MIC breakpoints recommended by the SRGA (S < or = 0.5 mg/L, R > or = 1 mg/L). Zone diameter histograms from KS, performed with a 50 microg disk, and from TE, using a 10 microg disk, showed a clustering of wild type strains around 41 mm and 30 mm, respectively, reflecting differences in methodology. Zone breakpoints calculated from the equations were validated by comparison with the histograms. Breakpoints were also calculated for a suggested lower disk content in Sweden, 10 microg, and validated in tests of clinical isolates and by histogram analysis. PMID- 12583436 TI - The influence of CD4 and CXCR4 on maedi-visna virus-induced syncytium formation. AB - CD4 is the principal binding site for human and simian immunodeficiency virus (HIV/SIV) receptor interactions and the a chemokine receptor CXCR4 has been implicated as a primordial lentivirus receptor. This study sought to determine the relevance of CD4 and CXCR4 in virus-receptor interactions for the prototype lentivirus, maedi-visna virus (MVV) of sheep. Neither CD4 nor alpha/beta chemokine receptors represent principal receptors for MVV since human osteosarcoma cells devoid of these molecules were susceptible to productive infection. Interestingly, the presence of either CD4 and/or CXCR4 on indicator cells dramatically enhanced MVV-induced cell fusion (syncytium formation) for three independent virus strains. Syncytium formation results from virus-receptor interactions and can be inhibited by receptor ligands. However, neither SDF-la that binds CXCR4 nor recombinant gp120 (rgp120) that binds CD4 could specifically inhibit the observed enhancement of MVV-induced cell fusion under conditions that significantly reduced HIV-1-induced cell fusion. Our observations suggest that CD4 and CXCR4 may represent optional auxiliary components of an MVV receptor (or receptor complex) that facilitate MVV-mediated membrane fusion events, a feature important for virus entry. This potential accessory role for CXCR4 in MW receptor interactions may reflect the distant relationship between the ovine (MVV) and the human/feline lentiviruses (HIV/FIV). PMID- 12583437 TI - Modes of participation of macrophages in the formation of membranocystic lesions. AB - We investigated the modes of participation of macrophages in the formation of non specific (i.e., unrelated to membranous lipodystrophy) membranocystic lesions (MCLs). Surgical specimens of atherosclerosis and chronic panniculitis containing MCLs as well as experimentally formed MCLs were examined, using paraffin sections, by Sudan black B (SBB) stain and by immunostaining for CD68, Bax protein and advanced glycation end products (AGEs). In the atherosclerotic lesions, MCLs were formed by a gradual fusion of SBB-positive multivesicular structures in the macrophages, associated with the cell breakdown. MCLs in the panniculitis were formed in the macrophages, as in atherosclerosis, or derived from degenerative fat cells, often with attachment of macrophages. CD68 was demonstrated in some MCLs or around them, and Bax was also present in some MCLs as well as some macrophages in both lesions. On the other hand, macrophages were not involved in the experimental MCLs. AGEs were widely detected in some MCLs, macrophages and degenerative background in all specimens. These results suggest that MCLs share AGEs as common components with various types of fat degeneration, and are formed physicochemically in the event that macrophages cannot dispose of the degenerative fat sufficiently due to their relative functional impairment and/or the physical circumstances. PMID- 12583438 TI - Gastric inverted hyperplastic polyp. Report of four cases and relation to gastritis cystica profunda. AB - Gastric inverted hyperplastic polyp (IHP) is a rare type of gastric polyp, and is characterized by downward growth of the hyperplastic mucosal components into the submucosa. To the best ofour knowledge, 16 gastric IHP cases have been described in the English literature, but the pathogenesis has not been established. We report the clinical and pathological findings of four gastric IHP cases. The lesions were mainly composed of hyperplastic foveolar-type glands with focal cystic dilatation. Pyloric type glands, endocrine cells, acinic cell metaplasia, and smooth muscle bundles were also seen as components of the polyp. Two cases (cases 1 and 4) coexisted with multifocal gastritis cystica profunda (GCP) and gastric adenocarcinoma. Case 4 furthermore exhibited an intermediate form between IHP and GCP. We suggest that IHP may be GCP associated with exaggeratedly hyperplastic and metaplastic changes. In case 4, the coexisting gastric carcinoma was mainly located in the submucosa, whilst the mucosal component was minimal. Five out of twenty reported gastric IHP cases, including our cases, coexisted with gastric adenocarcinoma. These facts would lead us to further investigate the relation between gastric IHP and carcinoma. PMID- 12583439 TI - Bax, Bcl-2, fas and Fas-L antigen expression in human seminoma: correlation with the apoptotic index. AB - Apoptosis plays a crucial role in the regulation of spermatogenesis in male germ cells and is, at least in part, modulated by Bcl-2, Bax, and the Fas pathway. Seminomas have a favourable outcome and respond to radio-/chemotherapy with an increased rate of apoptosis. The expression of Bax, Bcl-2, Fas and Fas-ligand (Fas-L) in human seminoma was evaluated and correlated with the apoptotic index. Twenty-nine classical seminomas were examined by immunohistochemistry and Western blotting using antibodies against Bax, Bcl-2, Fas and Fas-L. Apoptosis was detected by in-situ end-labeling of fragmented DNA and the apoptotic index (AI) was determined. Expression of Fas was found in 26 (89.7%) of Fas-L in 24 seminomas (82.2%); none of the tumours expressed Bcl-2. No correlation between the AI and Fas, Fas-L or Bcl-2 expression was found. Bax was demonstrated in 20/29 tumours (69%). Bax-positive tumours showed an increased AI of 4.75 +/- 2.38% in contrast to 2.60 +/- 1.23% of the Bax-negative tumours (P = 0.002). The number of Bax-positive tumour cells and apoptotic cells revealed a significant correlation using chi2-test (P = 0.04) and linear regression (r = 0.54, P = 0.001). Therefore, Bax seems to play a determinant role in the modulation of apoptosis in human seminoma that may be linked to a favourable outcome. PMID- 12583440 TI - Pheochromocytoma of the urinary bladder. AB - The case of a 65-year-old male with pheochromocytoma arising in the urinary bladder is presented. Clinical evaluation included ultrasonography, intravenous pyelography and computerized tomography (CT) scan. Transurethral resection of an exophytic tumor 1 cm in diameter was performed. The histological diagnosis was pheochromocytoma of the bladder. The evaluation and management of this type of tumor is discussed. PMID- 12583441 TI - Acute xenograft rejection, late xenograft rejection and long term survival xenografts in the hamster-to-rat heart transplantation model: histological characterisation under low-dose of FK506. AB - BACKGROUND: Long-term survival studies have been conducted in hamster-to-rat cardiac models with a range of immunosuppressive treatments, but the histological pattern of Late Xenograft Rejection (LXR) has not been outlined. This study offers a detailed description of the histological changes in cardiac xenografts under three different immunological responses. MATERIALS AND METHODS: Heterotopic hamster-to-Lewis rat cardiac transplant. Recipients were administered an antiproliferative drug (MMF, 25 mg/kg, or CyP, 10 mg/kg, from day -7 to +7 or from day 0 to +7, according to group) and FK506 (0.2 mg/kg; from day 0 to +30 or continuously). Unmodified recipients were used as controls. Conventional histology and indirect immunofluorescence of IgM, IgG and C3 deposits were performed. RESULTS: In our study, xenografted rats that did not receive treatment developed a pattern of Acute Xenograft Rejection (AXR), with substantial tissue breakdown. Pretreated and treated animals until day 30 post-transplant developed LXR that may present two different histological patterns: one with vascular damage and predominant interstitial haemorrhage, and the other with extensive myocardial fibrosis. Long-term surviving rats (LTS) showed a morphology that was almost normal, with mild fibrosis and vascular endothelium preserved. CONCLUSIONS: AXR, LXR and LTS in the hamster-to-rat heart transplantation model present a common humoral mechanism although their histopathological patterns are different depending on the length of immunosuppressive treatment but not on the type of antiproliferative drug administered. Pretreatment exerts an effect on fibrosis formation. PMID- 12583442 TI - Quality of fetal, perinatal and infant autopsy reports. An audit of all reports of postmortem examinations following fetal, perinatal and infant death in Rogaland County, Western Norway, 1997-1999. AB - The aim of the present study was to investigate the quality of fetal, perinatal and infant (< 12 months of age) autopsy reports. One hundred and thirty-six reports were scored for seven major categories (History, External description, Internal examination, Histology report, Other investigations, Placenta, Conclusions/Commentary) derived from "Guidelines for post mortem reports" published by the Royal College of Pathologists. The time taken for the final reports to be issued was also investigated. Of the 136 reports, 31 failed to reach a minimum standard of a 50% score. The one major category clearly falling below the recommended standard was Conclusions/Commentary. Other major categories such as History, External description and Histology report were also in need of improvement. Only three final reports were issued within the Department of Pathology's goal of 4 weeks. Although 77% of the reports fulfilled a minimum standard of a 50% score, the audit clearly showed that the standardised report form used was in need of revision and that many reports failed to give the clinicians an evaluation of the data presented in the autopsy reports. Based on the present findings, a number of measures have been implemented in order to improve the quality of future fetal, perinatal and infant autopsy reports. PMID- 12583443 TI - Elections and politics: the good, the bad, and the ugly. PMID- 12583444 TI - Membership increases one dentist at a time. PMID- 12583445 TI - HIV vaccine holds promise of global effectiveness. PMID- 12583446 TI - Paying off big debt fast: dentists share their stories. PMID- 12583447 TI - The music of our profession. PMID- 12583448 TI - A turn of the flywheel. PMID- 12583449 TI - CDA Code of Ethics. PMID- 12583450 TI - Gender-specific practice. PMID- 12583451 TI - Long-term HIV/AIDS survival estimation in the highly active antiretroviral therapy era. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) prolongs short-term survival in patients with HIV/AIDS. HAART has only been available since 1996; thus, no long-term survival data are available. Computer simulation models extrapolating short-term survival data can provide estimates of long-term survival. These survival estimates may assist patients and clinicians in HAART treatment planning. The authors construct a computer simulation model based on observational data to estimate long-term survival in a cohort of HIV/AIDS patients undergoing treatment with HAART. METHODS: The authors use data from the Collaboration in HIV Outcomes Research-US (CHORUS) observational cohort (N = 4791), the published literature, and US Life Tables to specify a computer simulation model of expected survival accounting for baseline CD4 cell count, progressive HAART treatment failure, progressive risk of HAART on treatment mortality, and age-associated mortality. Time to treatment failure for each of three rounds of HAART and risk of mortality on-treatment were estimated using parametric survival models with censoring of follow-up fit to CHORUS data. Off treatment survival after HAART failure was estimated from the pre-HAART literature. Age-associated mortality was taken from US Life Tables. RESULTS: Median projected survivals stratified by baseline CD4 cell count subgroups were CD4 > 200 cells/mm3, 15.4 years; CD4 < or = 200 cells/mm3, 8.5 years; and CD4 < or = 50 cells/mm3, 5.5 years. These values are 4 to 6 years longer than pre-HAART cohorts. The sensitivity analyses showed that the model survival predictions were most sensitive to the treatment failure rate, the on-treatment mortality rate, and the number of treatment rounds. CONCLUSIONS: Computer simulation modeling of long-term survival of patients with HIV/AIDS on HAART--accounting for differential treatment failure and death rates stratified by CD4 cell count and age-associated mortality--suggests a relatively consistent 4- to 6-year survival benefit over pre-HAART therapies. PMID- 12583452 TI - A comparison of Bayesian and maximum likelihood methods to determine the performance of a point of care test for Helicobacter pylori in the office setting. AB - OBJECTIVE: Evaluations of point of care tests (PCT) are often hampered by a lack of appropriate gold standards. This study aimed to compare the results of a Bayesian statistical analysis and a maximum likelihood method to evaluate the performance of a PCT for Helicobacter pylori in primary care. METHODS: The Helisal Rapid Blood Test (Cortecs Diagnostics) was performed in 311 patients from 6 primary care centers, and a concurrent venous sample was taken for 2 enzyme linked immunosorbent assays (ELISA) performed at the laboratory, blind to the PCT result. The Bayesian analysis was conducted using Markov Chain Monte Carlo methods (WinBUGS). The performance characteristics of the PCT and the 2 ELISA tests were estimated together with 95% credible intervals (95% CIs). RESULTS: The estimate of prevalence of H. pylori in this population was 64% (95% CI, 59% to 70%), the sensitivity and specificity of the PCT were 89% (84% to 94%) and 84% (77% to 91%), respectively (likelihood ratios positive 5.6, negative 0.13). The equivalent maximum likelihood results were prevalence, 65%; sensitivity, 90%; and specificity, 83%. CONCLUSIONS: The Helisal Rapid Blood Test performed as well as laboratory-based ELISA tests in this cohort of patients. The Bayesian analysis and the maximum likelihood method gave similar results, the Bayesian method also simultaneously estimating 95% CIs. PMID- 12583453 TI - The reliability of assessing the appropriateness of requested diagnostic tests. AB - Despite a poor reliability, peer assessment is the traditional method to assess the appropriateness of health care activities. This article describes the reliability of the human assessment of the appropriateness of diagnostic tests requests. The authors used a random selection of 1217 tests from 253 request forms submitted by general practitioners in the Maastricht region of The Netherlands. Three reviewers independently assessed the appropriateness of each requested test. Interrater kappa values ranged from 0.33 to 0.42, and kappa values of intrarater agreement ranged from 0.48 to 0.68. The joint reliability coefficient of the 3 reviewers was 0.66. This reliability is sufficient to review test ordering over a series of cases but is not sufficient to make case-by-case assessments. Sixteen reviewers are needed to obtain a joint reliability of 0.95. The authors conclude that there is substantial variation in assessment concerning what is an appropriately requested diagnostic test and that this feedback method is not reliable enough to make a case-by-case assessment. Computer support maybe beneficial to support and make the process of peer review more uniform. PMID- 12583454 TI - Use of Bayesian Markov Chain Monte Carlo methods to model cost-of-illness data. AB - It is well known that the modeling of cost data is often problematic due to the distribution of such data. Commonly observed problems include 1) a strongly right skewed data distribution and 2) a significant percentage of zero-cost observations. This article demonstrates how a hurdle model can be implemented from a Bayesian perspective by means of Markov Chain Monte Carlo simulation methods using the freely available software WinBUGS. Assessment of model fit is addressed through the implementation of two cross-validation methods. The relative merits of this Bayesian approach compared to the classical equivalent are discussed in detail. To illustrate the methods described, patient-specific non-health-care resource-use data from a prospective longitudinal study and the Norfolk Arthritis Register (NOAR) are utilized for 218 individuals with early inflammatory polyarthritis (IP). The NOAR database also includes information on various patient-level covariates. PMID- 12583455 TI - Model-based methodology for analyzing incomplete quality-of-life data and integrating them into the Q-TWiST framework. AB - BACKGROUND: The standard Q-TWiST approach defines a series of health states and weights each state's duration according to its quality of life (QOL) to calculate quality-adjusted lifetimes. However, a fixed weight may not adequately reflect time variations in QOL. METHODS: To account for measurements derived from irregular visits and informative missing data, the authors estimated the mean QOL profile using a mixed-effect growth curve model for the response, combined with a logistic regression model for the drop-out process. RESULTS: Using data from a clinical study of lymphoma patients, the authors demonstrated better readaptation to normal life for patients younger than 30. Sensitivity analyses and computer simulations demonstrated that modeling the drop-out probability as a function of the QOL measurements is necessary if conditioning by health state is not possible. CONCLUSION: Our model-based approach is useful to analyze studies with incomplete QOL data, especially when approximate QOL assessment by health state is not possible. PMID- 12583456 TI - Toward improved methods for measurement of utility: automated repair of errors in elicitations. AB - This study examines the effects of use of an automated computer protocol to correct a common error in utility elicitations--namely, scoring of a health state with a greater impairment as being more desirable than one with lesser impairment. The authors studied the protocol in a sample of 563 members of the Internet-using public. Results revealed that errors were common (17% of ratings) but were typically successfully repaired (individuals who made only 1 or 2 errors had a 75% chance of repairing them). The values of individuals who repaired errors were similar to those without apparent error. In contrast, individuals who refused to repair errors had lower scores for the best health state in the series and higher ones for the worst health state. Results suggest that the repair procedures were successful and that inclusion of utility scores from individuals who fail to repair illogically ordered ratings may bias estimates of mean utilities. PMID- 12583457 TI - Economic evaluation of vaccination programs: the impact of herd-immunity. AB - The unique characteristic of vaccination is that it not only reduces the incidence of disease in those immunized but also indirectly protects nonvaccinated susceptibles against infection (produces herd-immunity). The bulk of economic evaluations of vaccination programs continue to use models that cannot take into account the indirect effects produced by herd-immunity. Here, the authors illustrate the importance of incorporating herd-immunity externalities when assessing the cost-effectiveness of vaccination progams. To do this, they compare 2 methods of estimating the benefits of routine mass vaccination: one that includes herd-immunity (dynamic approach) and one that does not (static approach). Finally, they use the results to clarify a number of misconceptions that are common in the literature concerning herd-immunity and dynamical effects produced by models. PMID- 12583458 TI - SMDM presidential address: Hearsay or heresy--are health decision scientists too left brained? PMID- 12583459 TI - Law and ethics. Privacy and the idea/expression dichotomy. PMID- 12583460 TI - Primary IgA nephropathy-clinical course and outcome. PMID- 12583461 TI - Prognostic factors in immunoglobulin-A nephropathy. AB - AIM: To study the course, identify prognostic factors and a model predictive for development of end stage renal failure (ESRF) in adults with immunoglobulin-A nephropathy (IgA-N). DESIGN: Retrospective cohort. SETTING: Madras Medical College and Government General Hospital, Chennai, Tamil Nadu. PARTICIPANTS: Ninety eight adult patients with diagnosis of primary IgA nephropathy. RESULTS: Out of 98 patient 64 (65.3%) were men. Mean age of presentation was 25.7 years. The predominant renal lesions included nephrotic syndrome in 25 (25.5%), rapidly progressive renal failure and accelerated malignant hypertension in 21 (21.4%) each, chronic renal failure in 13 (13.3%), hypertension in nine (9.2%) haematuria in five (5.1%) and acute renal failure in four (4.1%). Sixty (61%) had renal failure at diagnosis. Age > 25 years, glomerular histology of Hass subclass V and interstitial fibrosis were significant factors. Forty (48.2%) (IR) patients developed ESRF during follow up. Serum creatinine > 5 mg/dl (hazard ratio: 5.37, 95% confidence interval CI 2.49-11.58) Hass-V (3.74, 1.60-8.76), crescents (4.08, 1.52-10.94) and IF (6.15, 2.0-19.0) were associated with disease progression in the multivariate analysis. Five years renal survival (CI) was 38.5% (24.6% 52.3%). Among those with no risk factor, 95% had not reached ESRF by 42 months. The median ESRF-free survival with one, two and three or four risk factors was 65, 16 and four months, respectively. CONCLUSIONS: Serum creatinine > 5 mg/dl, crescents: Hass-V histology and interstitial fibrosis predict the progression to ESRF. Renal biopsies should include a detailed assessment of the tubulointerstitium. PMID- 12583462 TI - Clinical profile of hereditary spherocytosis in North India. AB - AIMS OF THE STUDY: Hereditary spherocytosis (HS) is a familial hemolytic disorder manifesting as anaemia, recurrent jaundice, splenomegaly with marked heterogeneity in clinical presentation. The objective was to study the clinical spectrum of the disorder in India. METHODOLOGY: We studied 50 HS patients and followed them for up to six years (Age range 2-47 years). RESULTS: The presenting features were jaundice 35 out of 50, anaemia 30 out of 50 (requiring blood transfusion in 25). Splenomegaly was found in all patients. Increased osmotic fragility was found in all patients whereas spherocytes were found in only 19 out of 42 patients. Reduced red cell survival was noted in 9/12 patients studied with 51Cr labeled RBCs. There was a definite improvement in the hemoglobin values in those who underwent splenectomy. Thirteen cases had similarly affected family member/s. Fifteen of the cases had family history consistent with autosomal dominant (AD) inheritance (eight families) while in six cases (5 families), inheritance was likely to be autosomal recessive (AR). There was intrafamilial variability in the age of presentation in the AD families. CONCLUSIONS: Our results suggest that both autosomal dominant and recessive patterns of HS are seen in India and the clinical profile of the Indian HS patients is similar to that described in other populations. HS presenting in childhood is also not uncommon. However, the predominant underlying protein defect in Indian patients needs to be characterized. PMID- 12583463 TI - Clinical profile of sickle cell trait. AB - OBJECTIVES: Although sickle cell trait is considered a harmless condition in ordinary circumstances, a large number of pathological conditions have been attributed to it often without a scientific basis. Many patients visit this centre with various complaints and on testing are found to be sickle cell trait. Hence it was thought necessary to analyse these cases to find out the nature of their ailments. METHODS: Two hundred cases of sickle cell trait diagnosed by sickling test and hemoglobin electrophoresis on CAM, and 150 age and sex matched control cases with normal hemoglobin phenotype from a survey were studied. Hemoglobin estimation was done in all by cyanmethemoglobin method. Besides history and clinical examination other relevant investigations were done as necessary to arrive at the diagnosis. Seven cases of sickle cell trait were asymptomatic while the rest were suffering from different conditions. 51% of sickle cell trait and 86% of control cases had mild to severe anaemia which improved with iron therapy in trait cases. Hepatomegaly (11% vs 4.6%), epistaxis (5% vs 0.66%) and cholelithiasis (3% vs 0%) was seen in significantly more number in sickle cell traits than the control cases. Splenomegaly and midsystolic murmurs were present in higher percent of cases but was not statistically significant. There was one case of epilepsy with multiple small infarcts in the brain and another with focal fits with epileptogenic focus in the left cerebral hemisphere where no other cause could be found except sickle cell trait. CONCLUSION: The ailments of sickle cell trait cases are like persons with normal hemoglobin. Anaemia is not more frequent and can improve with iron therapy. However, hepatomegaly, epistaxis, cholelythiasis are seen more frequently and minor cerebral infarcts probably can occur in sickle cell trait. These require more elaborate studies to decide their pathogensis. PMID- 12583464 TI - Oxidant stress in pre-eclampsia and essential hypertension. AB - BACKGROUND: Endothelial cell dysfunction may play a role in the pathobiology of pre-eclampsia and human essential hypertension. Vasodilators and platelet anti aggregators such as prostacyclin and nitric oxide are produced by endothelial cells. The half-life of prostacyclin and nitric oxide are reduced by superoxide anion, whereas superoxide dismutase antagonizes its action. OBJECTIVES: To estimate the plasma concentrations of nitric oxide and lipid peroxides and those of catalase and superoxide dismutase in patients with pre-eclampsia and essential hypertension. METHODS: Patients of essential hypertension and pre-eclampsia were selected for the study. Nitric oxide and lipid peroxides were estimated in the plasma and anti-oxidants catalase and superoxide dismutase were estimated in the RBC membranes. RESULTS: The ratio between lipid peroxides and nitric oxide was elevated and the activity of superoxide dismutase reduced in patients with pre eclampsia and uncontrolled essential hypertension. CONCLUSION: These results suggest that oxidants and anti-oxidants are altered in human essential hypertension and pre-eclampsia. PMID- 12583465 TI - Health conditions in the tribal villages of South Bihar: an epidemiologic survey. AB - In order to understand and establish an effective approach toward sustainable rural health development in the tribal region of South Bihar, an extensive socio medical survey on over 90% of the houses of four tribal villages was undertaken. Using both a family-based questionnaire and screening physical examinations, the vital statistics, disease prevalence rates, immunization rates, and use of maternal health services in this neglected and previously unstudied area were determined. Local health-related behaviours and attitudes as well as nutritional status and family dietary intake were also focused upon. The results indicate that disease prevalence and mortality rates are significantly higher than expected and that existing services are less effective than previously thought. More importantly, the survey offers evidence that the fundamental obstacle toward improved community health remains a vast lack of health awareness and education in the villages; specifically in the areas of nutrition, immunization, diarrhoea prevention and treatment, maternity care, and family planning. Surveys of this kind provide vital information to health professionals working in underserved areas in India and should be an integral part of any such developmental undertaking. PMID- 12583466 TI - Evaluation of ventilator-associated lower respiratory tract infection and tracheobronchial aspiration of gastrointestinal contents. AB - STUDY OBJECTIVE: Ventilator-associated lower respiratory tract infection (LRTI) in a set up of respiratory intensive care unit was evaluated. Also the incidence of tracheobronchial aspiration of gastrointestinal contents was explored among these patients. DESIGN: Twenty eight patients admitted in the respiratory intensive care unit of referral teaching hospital were studied. All patients were evaluated for tracheobronchial aspiration of gastrointestinal contents in the initial 48 hours of initiating mechanical ventilation using methylene blue as marker and glucostix reagent strip. Protected endotracheal aspirate was secured after 48 hours of beginning of mechanical ventilation and was submitted for microbiological assay. RESULTS: Of the 28 mechanically ventilated patients, 82% were found to have developed LRTI. Aerobic gram-negative bacilli accounted for 79% of the infections, Klesiella responsible for 39% while Pseudomonas and Escherichia accounted for 18% each. The remaining 4% were caused by Acinetobacter. Gram-positive cocci were responsible for 21% of infection with Staphylococcus and enterococci 14% and 7% respectively. Tracheobronchial aspiration of GI contents was found to be present in 72% of the patients which was significantly correlated with the incidence of LRTI (p=0.014). Significant association was also found between the incidence of LRTI and mortality (p=0.05). CONCLUSION: The incidence of gastrointestinal aspiration was found to be high and correlation with LRTI was significant. Both glucostix reagent strip and methylene blue were observed to have same positivity rate. PMID- 12583467 TI - Prevalence of hepatitis B infection within family contacts of chronic liver disease patients--does HBeAg positivity really matter? AB - BACKGROUND: The risk of infectivity is known to be high in contacts of HBeAg positive chronically infected patient. We investigated and compared the frequency and significance of transmission of HBV infection from chronic liver disease patients (CLD) with HBeAg or anti-HBe and HBV DNA positive status. MATERIAL AND METHODS: Four hundred and seventy nine contacts [first degree blood relatives (n=278), second degree contacts (n=139) and sexual contacts (n=62)] of 92 HBV related, liver biopsy proven, CLD patients were studied. Three hundred and seventy three belonged to 65 index patients with HBsAg+ve, HBeAg+ve, HBV DNA+ve, HBV DNA+ve infection and 106 belonged to 27 index patients with (HBsAg+ve, HBeAg ve, anti-HBe+ve, HBV DNA+ve infection). One hundred and seventy six family members, age and sex matched, belonging to 38 healthy individuals, with no history of liver disease or HBV positivity, served as controls. Viral serology and quantitative DNA estimation was done in index patients. RESULTS: Forty nine of 65 (75.4%) families of HBeAg+ve and 63% families of HBeAg-ve index patients had one or more family member exposed to HBV (positive family, p=ns). The chronic HBV infection (HBsAg+ve) and past-exposure (only IgG anti-HBc+ve) rates in the contacts of HBeAg+ve and HBeAg-ve index patients were 17.4% and 19.8% (p=ns), and 31% and 14.2% respectively, both being significantly higher (P < 0.01) than the prevalence rates in the control group (chronic HBV infection 2.3%, past-exposure 10.2%). Overall, 48.5% and 34% (p < 0.05) of contacts in the HBeAg+ve and HBeAg ve groups had markers of HBV infection. The quantitative HBV DNA levels were comparable between HBeAg+ve and HBeAg-ve index patients (1712 +/- 356 pg/ml vs 1802 +/- 812 pg/ml). First degree relatives had higher chronic HBV infection rates than second degree contacts (29% vs. 0%, p < 0.05). The duration of symptomatic illness of HBeAg+ve index patients was longer than HBeAg-ve (p < 0.05). A significant proportion of HBsAg+ve first degree relatives of HBeAg+ve (33%) and HBeAg-ve (40%) patients, had evidence of CLD. CONCLUSIONS: (i) The frequency of transmission of HBV infection is nearly similar in contacts of HBeAg+ve and HBeAg-ve infected patients, more so in first degree relatives, (ii) these observations make family contacts a very high risk group, requiring priority screening and vaccination against HBV. PMID- 12583468 TI - Ethics in genetic research. AB - Studies of genetic variation should be conducted in developing countries only after community consultation where identifiable leadership exists and with the individual consent. Studies of particular genetic diseases require the consent of the individual, community consultation may be appropriate, but should not pre empt the interests of the individuals at risk. Gene therapy trials should not be conducted in the developing world, at least until such therapy becomes effective and relatively inexpensive. Results of genetic tests should be provided to subjects only if the test has been demonstrated to have sufficient clinical validity. Results should never be disclosed to relatives, except as may be unavoidable in the context of pedigree research. Policies regarding disclosure of test results should be included in the informed consent process. PMID- 12583469 TI - The clinical spectrum of neurocysticercosis in the Uttaranchal region. AB - OBJECTIVES: To study the clinical and radiological spectrum of neurocysticercosis in the Uttaranchal region. METHODS: A total of 592 patients of neurocysticercosis attending various health camps, medical outdoor and indoor department were studied for their clinical and radiological presentation. RESULTS: Out of the total 15,000 patients screened, 592 patients (3.94%) were found to be suffering from neurocysticercosis. Majority of them were in the age group of 21-30 years. Most of them presented with generalized seizures (62.5%), others had headache (37.5%), focal seizures (20.8%), localizing neurological signs (16.6%), meningitis (4.1%), and dementia (1.3%). Radiologically, parenchymal lesions were most prevalent (81%). However, interventricular lesions were also not very uncommon (32%). Most of them had single lesions (53.5%), 26.7% had multiple lesions and 0.7% had midline shift. Significantly, large number of patients were vegetarians (39%); only 17.6% were pork eaters while the remaining 82.4% never had pork in their meals. CONCLUSION: We believe that such high percentage is at least a rough indicator of high incidence and prevalence of neurocysticercosis in the Uttaranchal region. They had wide range of clinical and radiological presentation. Small number of pork eaters and a significant number of vegetarians suggest other modes of transmission and needs further evaluation. PMID- 12583470 TI - W M Haffkine (1860-1930) postal stamps released in India-1960 & Israel-1993. PMID- 12583471 TI - Pictorial CME. A racemose arteriovenous malformation characterized by dilatation and tortuosity of retinal vessels. PMID- 12583472 TI - Attitude of teachers towards teaching. AB - INTRODUCTION: Teaching is an art and the quality of teaching depends on the love, dedication and devotion of the teacher towards the subject of the knowledge. The quality of any teaching programme cannot rise above the quality of its teachers. In medical colleges it is the teacher who is responsible for influencing a student's learning of the subject. METHODS: We assessed the attitude of 31 teachers working at MGIMS. Twenty one of them were of the rank of Reader and above and had more than five years teaching experience. Ten were of the rank of lecturers with three years of teaching experience. The assessment was done by a likert type scale containing 20 items on various aspects of teaching. All the participants were given the scale and requested to mark the agreement or otherwise on a scale i.e. strongly disagree, disagree, cannot say, disagree, strongly agree. OBSERVATION AND CONCLUSION: The mean score was 3.808, which indicates a positive attitude. There was not much difference in attitude of teachers in different group. Thus indicating that our study group has predominantly positive attitude for most of the items. This positive attitude helps the teachers to be role model for the future generation of students. PMID- 12583473 TI - Surfactant protein-A in lung lavage fluid obtained from patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: Alterations in surface-active material may contribute to the pathogenesis of idiopathic pulmonary fibrosis both by increasing the elastic recoil due to surface forces and by promoting alveolar collapse. This study intends to evaluate the surfactant protein-A levels in bronchoalveolar lavage in patients of idiopathic pulmonary fibrosis. MATERIAL AND METHODS: Ten patients of idiopathic pulmonary fibrosis (IPF) of either sex were taken up for estimation of surfactant protein-A (SP-A) levels in bronchoalveolar lavage. The final diagnosis of IPF was based on histopathology and high resolution CT. Ten controls were taken out of which five patients were histopathologically proven cases of sarcoidosis and five patients with normal chest X-ray and CT scan. History, physical examination and routine investigations were done to rule out any concomitant illness. RESULTS: Age range of patients varied from 37-65 years (mean +/- SD of 50.65 +/- 9.05) and controls 30-62 years (42.50 +/- 9.95). Fiberoptic bronchoscopy and bronchoalveolar lavage was carried out and fluid aspirated was studied for surfactant protein-A. The level of surfactant protein-A recovered was lower in patients with idiopathic pulmonary fibrosis in comparison to control group p < 0.01 (1.86 +/- 1.26 and 5.76 +/- 2.0 microg/ml respectively). The levels of surfactant protein-A also revealed that the level decreased with derangement of pulmonary function. CONCLUSIONS: The study thus showed that the level of surfactant protein A is reduced in patients with idiopathic pulmonary fibrosis. It is likely that they have a significant role in pathogenesis of idiopathic pulmonary fibrosis and its progression. Surfactant-A may have important therapeutic implications. Further studies are required for a definite answer. PMID- 12583474 TI - Emphysematous pyelonephritis. AB - Emphysematous pyelonephritis is a rare life-threatening suppurative necrotising infection of the renal parenchyma and perirenal tissue producing abscess formation and intrarenal gas. It is almost always seen in diabetics, making it a concern of the physician. A high index of suspicion is required for diagnosis and a combined approach has significantly reduced the mortality. PMID- 12583475 TI - New insights into circadian aspects of health and disease. AB - Early awakening and early to bed as well as good conduct, thought, diet, interpersonal dealings and physical activity have been suggested for healthy life in Ayurveda. Circadian rhythms, coordinated in part by the parietal hypothalamic pituitary and adrenal mechanisms, have been reported in almost all variables examined thus far, including the circulation. It is possible that all metabolic functions undergo circadian rhythms. It remains to be explored whether these rhythms may be optimized by Ayurvedic practices. The onsets of myocardial ischemia, unstable angina, acute myocardial infarction, sudden cardiac death, and strokes have been reported to exhibit a circadian variation, with increased frequency in the second quarter of the day. An increased physical and mental load caused by an attempt to prepare for the day may be important in triggering acute cardiovascular events. Depending on their timing, meditation (Ayurvedic practice), n-3 fatty acids, coenzyme Q10, beta-blockers and estrogens may have beneficial effects, whereas progestins and mental load may have adverse effects on heart rate and blood pressure variability, which may be expressed by different circadian patterns. Around the clock serial recordings of blood pressure and heart rate during usual activities and lifestyles may be recorded and may be analyzed by computer-implemented curve fitting to assess the about 24-hour (circadian) variation, among other rhythmic, chaotic, and trend components of the time structure (chronome) of these variables. The new disease risk syndrome circadian hyper-amplitude-tension (CHAT), a condition characterized by an excessive circadian amplitude of blood pressure, cannot be ascertained on the basis of single casual blood pressure measurements. The International Chronome Ecological Study of Heart Rate (and blood pressure) Variability in various ethnic groups aims at collecting further evidence regarding the role of blood pressure and heart rate variability in the pathogenesis of cardiovascular events, while also examining any role played by ethnicity and lifestyle-associated factors. PMID- 12583476 TI - Acute paraplegia following viper bite. AB - Neurological complications following viper bite are uncommon and are generally as a result of intracerebral or subarachnoid bleed and rarely due to cerebral infarction. We report a young male who following viperine bite developed local tissue swelling, haemorrhagic manifestations due to disseminated intravascular coagulation and later developed acute flaccid paraplegia as a result of dorsal spinal cord involvement. PMID- 12583477 TI - Elephantiasic pretibial myxedema in Graves' disease. PMID- 12583478 TI - Depression--an adverse event with nifedipine. AB - Two cases are reported in which depression was associated with the use of calcium channel blocker, nifedipine. In one instance, a patient became unresponsive to treatment with nortriptyline when nifedipine was introduced. In both cases, the depression resolved following discontinuation of nifedipine. PMID- 12583479 TI - Acute myeloid leukemia presenting as splenic rupture. AB - We describe a rare case of acute myeloid leukemia presenting primarily as an acute abdomen due to spontaneous splenic rupture in a 19 years male patient. He was treated with splenectomy after failure of conservative management for splenic preservation but later succumbed to an intracerebral haemorrhage. PMID- 12583480 TI - Melioidosis--a report from Pondicherry, South India. AB - Melioidosis is an acute infectious disease caused by a safety-pin-shaped gram negative bacteria called Burkholderia pseudomallei. Here, we report the first case of melioidosis in a middle aged male agricultural worker, from Pondicherry. The isolation of this organism from subcutaneous nodules on the extensor aspect of his limbs underlines the diversity of its clinical presentation. Difficulty in identifying the organism which mimics any other non-fementing gram-negative bacilli (NFGNB) on cursory examination, highlights the importance of identification of NFGNB in endemic areas for specific treatment and prevention. PMID- 12583481 TI - Does cyclophosphamide benefit relapsing thrombotic thrombocytopenic purpura? AB - Thrombotic thrombocytopenic purpura (TTP) is a rare disease. A high index of suspicion is required for early diagnosis and treatment. It can present in acute as well as chronic forms. Cyclophosphamide has been reported to be beneficial in relapsing TTP. Herein a case of acute TTP evolving into relapsing phase of the disease is presented. Cyclophosphamide in our case achieved partial remission without the need for plasma exchange therapy. The rationale of cyclophosphamide in management of relapsing TTP is discussed. PMID- 12583482 TI - Virtual bronchoscopy in carcinoma of lung. AB - A 55 years female presented with a malignant pleural effusion. Post-paracentesis, a lobular mass was seen which was biopsied after virtual bronchoscopy. PMID- 12583483 TI - Successful outcome of pregnancy in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis is typically seen in older individuals. Occurrence of pregnancy in idiopathic pulmonary fibrosis is uncommon and ends with abortion in case it happens. Here we report a case of idiopathic pulmonary fibrosis presenting at 38 weeks of pregnancy who successfully delivered a male child without any significant complications. The disease was active at presentation and early postpartum period. PMID- 12583484 TI - Paroxysmal dyskinesia in phenytoin toxicity. PMID- 12583485 TI - Computed tomography as a diagnostic tool in acute renal cortical necrosis. PMID- 12583486 TI - Hepatic encephalopathy associated with central pontine myelinolysis. PMID- 12583487 TI - Cockayne syndrome. PMID- 12583488 TI - Prevalence of migraine in hypothyroidism. PMID- 12583489 TI - Postprandial hyperglycaemia--the real challenge in diabetes. PMID- 12583490 TI - Hepatotoxic reaction to warfarin in a recovering hepatitis patient with hypoalbuminenia. PMID- 12583491 TI - Boerhaave's syndrome in pregnancy. PMID- 12583492 TI - Post-prandial hyperglycemia--the real challenge in diabetes. PMID- 12583493 TI - Isolated follicle-stimulating hormone (FSH) deficiency syndrome (IFDS). PMID- 12583494 TI - Use of lidocaine spray for pain relief and improved quality of life in terminally ill cancer patients. PMID- 12583495 TI - Free radical activity in diabetic patients with and without microalbuminuria. PMID- 12583496 TI - Systematic literature reviews. PMID- 12583497 TI - Survey of nursing and medical profile prior to deinstitutionalization of a population with profound mental retardation. AB - During the planning stages of deinstitutionalization, the importance of nursing services must be recognized and given priority consideration in the community placement of persons with serious developmental disabilities. The objective of this study was to survey the medical and nursing profile of a group of nonambulatory, institutionalized individuals with profound mental retardation in anticipation of their nursing and medical needs in the community. Data were collected from the Individual Habilitation Plans of 55 individuals who had resided in a residential facility for individuals with mental retardation and were scheduled for community placement Serious medical problems in decreasing frequency were constipation (96%), seizure disorder (70%), poor dental hygiene (67%), cerebral palsy (62%), scoliosis (61%), contractions (41%), aspiration (44%), skin lesions (40%), and dysphagia (22%). Considering the complexity of health issues encountered in this population, adequate nursing and medical planning are critical to the wellness and successful community placement of a population with special needs. PMID- 12583498 TI - Couples' ratings of chronic obstructive pulmonary disease patients' quality of life. AB - The purpose of this study was to compare perceptual differences between chronic obstructive pulmonary disease (COPD) patients and their non-COPD spouses regarding the patients' health-related quality of life (HRQOL). Sixty-seven community-dwelling COPD patients and their non-COPD spouses participated. Participants completed the Sickness Impact Profile (SIP) and the St. George's Respiratory Questionnaire (SGRQ). On both questionnaires, spouses' perceptions of patients' HRQOL were lower than those of the patients themselves. At the bivariate level, statistically significant differences were found on the SIP in the physical domain and on the SGRQ in the psychosocial domain. At the multivariate level, the age and gender of both the patients and spouses accounted for 22% of score differences in the psychosocial domain of HRQOL for the SGRQ. Discordance was greatest among the oldest couples with female non-COPD spouses. Domain-specific HRQOL differences provide direction for supportive nursing interventions to enhance coping among COPD couples. PMID- 12583499 TI - Physiological and psychological markers associated with HIV-related fatigue. AB - Fatigue is among the most common and debilitating complaints of HIV-positive individuals. These data are part of a larger exploratory study investigating the relationships of selected psychological and physiological factors among 40 participants with HIV-related fatigue. Fatigue severity was measured using the HIV-Related Fatigue Scale. Fatigue was correlated with depression (r = .40, p < .01), state anxiety (r = .40, p <.01), and trait anxiety (r = .46, p <.01). Of the physiological factors, there were three statistically significant correlations: thyroid-stimulating hormone was negatively correlated with fatigue severity (r = -.36, p = .02), and platelets (r = .35, p = .03) and alkaline phosphatase (r = .27, p = .09) were positively correlated with fatigue severity. There were no correlations between fatigue severity and CD4 count (r = -.16, p = .31) or fatigue severity and HIV viral load levels (r = .031, p = .84). Even among the group with excellent viral suppression, fatigue scores were still very high for many participants. PMID- 12583500 TI - Self-management decision influences in heart failure: a preliminary investigation. AB - Self-management decision making places significant cognitive demands on individuals with heart failure (HF). Before theoretically valid decision supports can be developed, a better understanding of underlying decision characteristics is needed. The purpose of this preliminary study was to describe sources of self management decision influences for individuals (N = 38) with varying levels of HF (New York Heart Association (NYHA) classifications). Therapeutic value was the most influential source of self-management decision making for these patients, followed by personal beliefs and values, information sources, symptom influence, and task difficulty across all NYHA classes. Symptom influence was least influential for persons in Class II, whereas task difficulty was least influential for those in Classes III and IV. Moderate and significant associations were noted between information sources and personal beliefs and values and between information sources and symptom influence. As the disease progressed, a trend toward fewer decision influences, especially in the task difficulty scale, was noted. PMID- 12583501 TI - Acute dyspnea as perceived by patients with chronic obstructive pulmonary disease. AB - Like pain, dyspnea, or difficulty in breathing, is a strong subjective experience of physiologial distress. Using a model developed by DeVito, this study focused on the emotional aspects of the acute experience of dyspnea in patients diagnosed with chronic obstructive pulmonary disease (COPD) on a medical unit and explored nursing activities that eased the intensity of the symptoms. The study was descriptive in design and used a semistructured interview format with four open ended questions and a numerical self-assessment of dyspnea using the Modified 0 10 Borg Scale (MBS). The sample included 41 hospitalized patients with COPD between the ages of 43 and 89 years who had experienced episodes of acute dyspnea both prior to and during their current inpatient stays. This article provides descriptions of the experience of dyspnea, identifies themes from the participants' responses, notes the patients'perceptions of the efficacy of nursing actions, and extends the use of the MBS to an inpatient population. PMID- 12583502 TI - Symptom management and adjustment of patients with multiple sclerosis: a 4-year longitudinal intervention study. AB - The researchers studied the effectiveness of a nursing intervention in promoting adjustment and symptom management in individuals with multiple sclerosis (MS). This was a 4-year longitudinal study to determine whether the 4-week intensive outpatient program was effective in increasing adjustment to MS and if the treatment effect would last over time. A sample of 27 individuals with MS participated in the study. Treatment participants had significant improvements in symptom management at the 4-year follow up. This improvement was attributable to signficant improvements in sleep and fatigue levels. Although adjustment and self efficacy scores improved in the treatment group over time, this improvement was not superior to the control group. This was anticipated because the behavioral changes would precede improvement in adjustment to life following the diagnosis of MS. PMID- 12583503 TI - Mechanisms of reactivity of urinary bladder smooth muscle. AB - BACKGROUND: Mechanism of bladder smooth muscle contraction and relaxation and its pharmacological influence in various pathologic states are of incremental interest of investigators and clinicians. OBJECTIVES: The aim of our study was to assess the reactivity of urinary bladder smooth muscle in guinea pigs to two different pharmacological agents. METHODS: We compared the in vitro reactivity of smooth muscle strips of guinea pig urinary bladder to muscarinic stimulation by carbachol and acetylcholine. We prepared two kinds of strips, urothelium-denuded and with intact urothelium. Both kinds of strips were aerated under tension in Krebs-Henseleit's solution in organ-bath for 1 hour and after that cumulative concentration-response curves to carbachol (10(-9) - 10(-5) mol/l) and acetylcholine (10(-8) - 10(-3) mol/l) were constructed. RESULTS: We observed significantly higher reactivity of smooth muscle strips to carbachol, comparing to acetylcholine at the same concentrations both in strips with urothelium (at concentration 10(-5) mol/l: 22.1 g/100 mg vs 6.1 g/100 mg) and in urothelium denuded strips (at concentration 10(-5) mol/l: 24.5 g/100 mg vs 5.1 g/100 mg). The reactivity differences between strips with and without urothelium were not significant, however, in higher concentrations of acetylcholine (10(-4) and 10( 5) mol/l) and carbachol (10(-6) and 10(-5) mol/l) we noticed no significant inhibition of contractile response of smooth muscle strips with intact urothelium. CONCLUSION: In our experiments we confirmed that carbachol was more potent constrictor than acetylcholine in detrusor smooth muscle strips of guinea pigs. The presence of urothelium did not change the reactivity significantly. (Fig. 4, Ref: 23.). PMID- 12583504 TI - Levels of coenzyme Q10 in asthmatics. AB - BACKGROUND: The contribution of free oxygen radicals in the pathogenesis of bronchial asthma is generally accepted. The modulation of antioxidative defence by supplementation with antioxidants represents additive approach in complex management of the disease. The aim of the study was to assess the levels of coenzyme Q10, alpha-tocopherol, beta-carotene and malondialdehyde (end-stage parameter of lipid peroxidation) in asthmatics (As). METHODS: Fifty six As (15 males and 41 females) aged from 19 to 72 yrs (mean age 46 yrs) were enrolled into the study. The control group comprised of 25 healthy volunteers (16 males, 9 females) aged 25-50 years. RESULTS: Concentrations of CoQ10 and alpha-tocopherol, decresed significantly both in plasma and whole blood, compared with healthy volunteers (p < 0.009, p < 0.004; p < 0.035, p < 0.001, respectively). The level of MDA was elevated, but not statisticaly significantly. No changes were seen in beta-carotene levels. Positive correlation was found between concentrations of CoQ10 and alpha-tocopherol. CONCLUSION: Our results suggest possible contribution of suboptimal concentrations of CoQ10 on antioxidative dysbalance in As and provide rationale for its supplementation with clinical evaluation. (Tab. 2, Fig. 1, Ref. 39.). PMID- 12583505 TI - Thrombolytic therapy for patients with acute myocardial infarction. Causes increasing the probability of not receiving thrombolytic therapy. AB - Intravenous thrombolysis is the most accessible and the most common form of reperfusion therapy. The aim of this study was to identify demographic, clinical and electrocardiographic factors, which based on published data and in patients included in the project Audit of diagnostic and therapeutic procedures in patients with acute myocardial infarction (AUDIT), increased the probability of not receiving thrombolytic therapy. In order to maximize the impact of thrombolytic therapy to reduce the case fatality rate associated with an acute myocardial infarction, we review, which a number of studies provide evidence on the usage of thrombolytic therapy in elder, women, patients with diabetes mellitus, bundle-branch block and after stroke. (Fig. 10, Ref. 52.). PMID- 12583506 TI - Coxsackie viral infection and orofacial cleft. AB - The incidence of orofacial cleft (OC) in newborns was compared with the occurrence of virus-neutralizing antibodies to coxsackie viruses in the serum of newborns and their mothers. No significant difference was found when comparing the seropositivity rates between the group of patients and the control group of healthy newborns. If the patients were divided according to the place of residence however, marked differences occurred between the regions. The lowest incidence of both--coxsackie infection and OC was determined in the region of Bratislava and the highest in the region of Zilina. The explanation of these findings recquires a more detailed analysis of genetic background, social and hygienic status, style of life and other factors, known to influence the development of OC as multi-etiological developmental disorder. (Tab. 2, Fig. 4, Ref. 12.). PMID- 12583507 TI - The use of heart rate variability in cardiology. AB - Heart rate variability is the marker of the heart's response to the autonomic nervous system activity. The decrease in HRV is a clinical predictive factor of overall cardiac mortality, and especially that of arrhythmia complications in the population after infarction. The parameters most significantly used in practice include SDNN and HRV index. Twenty-four-hour measurements are of higher predictive value, and an increase in the positive predictive value can be accomplished by multifactorial stratification. HRV is most frequently combined with ejection fraction and baroreflex sensitivity. The possibilities of HRV assessment are interesting also in coincidence with heart failure, arterial hypertension and atrial fibrillation also after heart transplantation. (Tab. 2, Ref. 63.). PMID- 12583508 TI - Telomerase inhibitors in anticancer therapy: gossypol as a potential telomerase inhibitor. AB - Telomerase is a reverse transcriptase which helps to stabilize the length of telomeres by adding TTAGGG repeats onto the telomeres that enables unlimited division of the cell. Absence of telomerase activity causes replication senescence and cell death. Activity of telomerase was detected in embryonal, gonadal and cancer cells, but was not detected in adult somatic tissues with the exception of tissues containing stem cells. Telomerase inhibitors are being considered as potential anticancer drugs with the hope that the lack of telomerase expression in normal somatic cells would result in a higher specific therapy with fewer side effects than conventional chemotherapy. Because of high activity of telomerase in gonadal cells, we hypothesized that one of the drugs which selectively interfere with spermatogenesis may act like a telomerase inhibitor. Gossypol is a male contraceptive derived from cotton seeds. It causes suppression of spermatogenesis by an unknown mechanism. It has a non-endocrine inhibiting effect on spermatogenesis, gradual onset of action, and irreversible suppression of spermatogenesis with increasing length of use. It also has an antiproliferative effect on cell lines derived from tumors, mild antineoplastic action in vivo with few side effects. It has mild inhibiting influence on the replication of HIV via possible inhibition of reverse transcriptase HIV. This shows its possible telomerase inhibiting activity. It appears that gossypol is a potential new drug for testicular cancer therapy, because of its spermatotoxic and cytotoxic activity on cell lines derivided from testicular tumors. We posit that the use of gossypol in combination with other anticancer chemotherapeutics can lead to a more effective therapy of human tumors. (Ref. 47.). PMID- 12583509 TI - Results and perspectives of the treatment of skeletal neoplasms. AB - Improvements in the therapy of skeletal neoplasms at the end of the 20th century are the reason for a retrospective view on the development of the therapy of skeletal neoplasms, especially in the Slovak Republic. Up to the sixties years of the 20th century was amputation the imperative in the surgical treatment of malignant tumors, above all tumors of the extremities. Then chemotherapy was introduced into the treatment of these tumors, in Slovak Republic it was later. Bone transplantations were widely used in the 70 and 80 years of 20th century. Autotransplantations of bone grafts were used at the Orthopaedic department in Bratislava already in the year 1953, but low number of appropriate grafts limited their use in clinical practice. In the 70 and 80 years development of bone banks was observed. Results of the treatment of malignant diseases with this technique are considered poor. Only few patients survived more than 5 years after the surgery, mainly due to inappropriate techniques of detection, localization and determination of the size of the tumor. Biopsy played above all at this time an important role in the diagnostics of bone tumors. Apart from basic examinations of the fixed material, processed after it was imbedded in particular medium, perioperative biopsy is recently being performed with the technique of frozen sections from unfixed tissue obtained during the surgery. This type of material processing prolongs the period from obtaining the material till making the diagnosis, but the advantages of this type of examination are undoubted. (Ref. 17.). PMID- 12583510 TI - The incidence and the characteristics of heart failure patients at a large medical department. AB - INTRODUCTION: Chronic heart failure (HF) has a prevalence of 6-10 % in patients above the age of 65 years, leading to high mortality, morbidity and re hospitalization. AIM: To analyse the incidence and the characteristics of patients with HF in a medical department. PATIENTS AND METHODS: 941 patients (males 424, females 517 with an average age of 64 years and 67 years respectively), hospitalized from 1.9.1998 to 28.2.1999 at the 1st Internal Department of the University Hospital. A retrospective analysis of their files: age, diagnosis, duration of hospitalization, and in those patients with HF: the occurrence of risk factors and clinical events( stroke, myocardial infarction, and atrial fibrillation). RESULTS: We have analyzed the incidence of diseases at a medical department where 58.7 % of patients suffered from cardiovascular diseases. Patients with HF represented 13.9 % of all hospitalized patients. Among the risk factors dominated hypertension, diabetes, obesity, hyperlipidemia and smoking. The average duration of hospitalization was 12.66 days in men and 13.67 days in women. The hospitalization period was 15.6 days in hypertensive diabetic males whereas in females it was 15.3 days. These patients were hospitalized in their 5th decade with the incidence of HF increasing with age. Their mortality was 11.5 % while the overall mortality was 9.7 % among all patients hospitalized. Those patients who died suffered from poly-morbidity. CONCLUSION: The prevalence of HF in our patients was 13.9 %. This value is higher than the data from literature. These patients suffered from poly-morbidity (hypertension: 53-75 %, diabetes: 30 %, obesity: 33-38 %, hyperlipidemia: 20-50 %, smoking: 11-32 %, myocardial infarction: 41-50 %) and had a high mortality. (Tab. 5, Fig. 2, Ref. 7.). PMID- 12583511 TI - Developing strategies for long-term follow up of cancer survivors. AB - The anticancer treatment has been increasingly successful over the past three decades. With current treatment, over two-thirds of children with cancer and more than half of adult cancer patients can be expected to survive their disease. As a result, there now exists a rapidly growing population of long-term survivors with increased risk of premature morbidity and mortality-induced by therapy. It has become apparent that these patients often face complex of organ damages and psychosocial problems. The awareness of the possible long-term complications of therapy is important for optimal health care for the current survivors and also for modifying future treatment regimens to avoid therapy that is associated with unacceptable morbidity and mortality. ((Tab. 1, Fig. 1, Ref. 17.). PMID- 12583512 TI - New trends in myofascial pain syndrome. AB - This review article summarizes recent studies on myofascial trigger point (MTrP) to further clarify the mechanism of MTrP. MTrP is the major cause of muscle pain (myofascial pain) in clinical practice. There are multiple MTrP loci in an MTrP region. An MTrP locus contains a sensory component (sensitive locus) and a motor component (active locus). A sensitive locus is the site from which pain, referred pain (ReP), and local twitch response (LTR) can be elicited by needle stimulation. Sensitive loci are probably sensitized nociceptors based on a histological study. They are widely distributed in the whole muscle, but are concentrated in the endplate zone. An active locus is the site from which spontaneous electrical activity (SEA) can be recorded. Active loci are dysfunctional endplates since SEA is essentially the same as endplate noise (EPN) recorded from an abnormal endplate as reported by neurophysiologists. Both ReP and LTRs are mediated through spinal cord mechanisms, demonstrated in both human and animal studies. The pathogenesis of MTrPs appears to be related to the integration in the spinal cord (formation of MTrP circuits) in response to the disturbance of the nerve endings and abnormal contractile mechanism at multiple dysfunctional endplates. Methods usually applied to treat MTrPs include stretch, massage, thermotherapy, electrotherapy, laser therapy, MTrP injection, dry needling, and acupuncture. The mechanism of acupuncture is similar to dry needling or MTrP injection. The new technique of MTrP injection can also be used to treat neurogenic spasticity. PMID- 12583513 TI - Should we perform direct stenting in selective patients? PMID- 12583514 TI - The feasibility of cartilage graft for mastoid obliteration in cholesteatoma surgery. PMID- 12583515 TI - Long-term clinical follow-up after successful direct coronary stenting without predilatation. AB - BACKGROUND: Direct stent implantation without predilatation is considered a promising new technique that may reduce procedural time, radiation exposure, ischemic time and cost, but little information is available concerning the long term outcome. The aim of this study was to investigate the long-term clinical outcome of successful direct stenting without predilatation. METHODS: We prospectively undertook a clinical follow-up program (minimum 8 months) in a consecutive series of 101 patients (113 lesions) who were successfully treated with direct stenting without predilatation. RESULTS: Clinical follow-up was obtained in all 101 patients at a mean period of 12.8 months (range 8 to 18.9). Stress test results were available in 94 patients (94%). During the follow-up period, 23 patients (23%) had one or more events, which included death in 2 patients (2%), target vessel revascularization in 14 (14%), myocardial infarction in 1 (1%) and positive stress test results or recurrence of symptoms (Canadian Cardiovascular Society I to II) treated medically in 6 (6%). Cumulative event free survival at 8 and 18 months were 80% and 72%, respectively. Long-term clinical event rate was not significantly different among the clinical presentations, lesion types, or stent types. Angiographic follow-up was performed in 43 (43%) patients with 45 lesions. Restenosis (defined as 50% diameter stenosis) was observed in 14 of the lesions (31%). CONCLUSIONS: Direct stenting without predilatation is an effective method of coronary intervention in terms of low long-term clinical event rate. PMID- 12583516 TI - Mastoid-obliteration surgery with cartilage for suppurative cholesteatomatous ears. AB - BACKGROUND: Autologous cartilage has been widely used as a material for tympanoplasty and mastoid-obliteration surgery. Since it remains vulnerable to an infectious condition, this study aims to evaluate if it can be used for a chronic suppurative cholesteatomatous ear to achieve single-stage tympanoplasty with mastoidectomy and obliteration of the cavity. METHODS: From January 1988 to August 1998, the records of patients with cholesteatoma who received cartilage obliteration surgery performed by Dr. Lien were examined for any infection associated problems within three months after the operation. The Fisher's exact test was used to compare the difference in the incidence of post-operative infection-associated problems between suppurative ears and non-suppurative ears. RESULTS: One hundred and seven surgical procedures performed upon 96 patients were included in this study, with 97 being primary surgical procedures and ten being revision surgery for previous open cavity with cholesteatoma recurrences. The average follow-up was 47.7 months. At time of surgery, there were 59 non suppurative ears; 48 ears were suppurative and cultured. Granulations were found in the middle ear or mastoid for 42 ears (39.3%). Five ears (4.7%) exhibited infection-associated post-operative problems within three post-operative months. Three cases (5.1%) were non-suppurative ears and two cases (4.2%) were suppurative ears. There was no significant statistical difference when examined with the Fisher's Exact test. CONCLUSIONS: In an immunocompetent patient with a cholesteatomatous chronic suppurative ear, autologous cartilage could potentially tolerate an infection condition to serve as a material for mastoid-obliteration in a single-stage surgical procedure. PMID- 12583517 TI - Transnasal sphenopalatine artery electrocautery for posterior epistaxis. AB - BACKGROUND: To evaluate the effects of sphenopalatine artery electrocautery for the treatment of recurrent posterior epistaxis. METHODS: Nine patients were enrolled in the study. Seven patients had hypertension, two had diabetes, two received irradiation for nasopharyngeal carcinoma, one had congestive heart failure and one was a heavy drinker. Three patients needed blood transfusion for profound blood loss. The sphenopalatine artery electrocautery was performed transnasally with endoscope. After the sphenopalatine foramen was identified through dissecting the mucosa of middle meatus posteriorly one centimeter from the choana, the neurovascular bundle in the sphenopalatine foramen was cauterized. RESULTS: Most operations can be performed within fifty minutes with minimal blood loss under local anesthesia. One patient developed minor epistaxis 2 months after surgery, and the bleeding was controlled with medical treatment. The other patients had no recurrent epistaxis after surgery. Thus, epistaxis was well controlled in all patients without complications. The follow-up duration was 2 to 14 months after surgery, with a mean duration of 10 months. CONCLUSIONS: Transnasal sphenopalatine artery electrocautery is a simple, effective and safe method for the treatment of posterior epistaxis. PMID- 12583518 TI - Risk factors for endogenous endophthalmitis secondary to Klebsiella pneumoniae liver abscess. AB - BACKGROUND: To identify the risk factors for endogenous endophthalmitis secondary to Klebsiella pneumoniae (KP) liver abscess. METHODS: Retrospective chart review of 200 patients with KP liver abscess from 1990 to 2000 was performed. Data variables included age, sex, past history, systemic condition, initial and final visual acuity, slit lamp biomicroscopy, intraocular pressure, fundus, course and treatment. RESULTS: Extrahepatic metastasis developed in 44 patients, including 18 cases (23 eyes) of endophthalmitis. Seventeen eyes had final vision less than counting fingers. Diabetes had significant association with the development of extrahepatic metastasis (p = 0.045) and the poor visual outcome of endophthalmitis (p = 0.022), whereas, neither the initial vision nor vitrectomy was significantly related to the outcome of endophthalmitis. CONCLUSIONS: The visual prognosis of endogenous endophthalmitis secondary to KP liver abscess is generally poor, and diabetes might be a significant risk factor for the poor visual outcome. Even when the initial vision is as bad as light perception only, some eyes can be saved. Both internists and ophthalmologists should be alert enough to detect and treat these patients early. PMID- 12583519 TI - Guillain-Barre syndrome: a retrospective, hospital-based study. AB - BACKGROUND: To obtain more information about Guillain-Barre syndrome (GBS) for Taiwanese patients. METHODS: We retrospectively reviewed the charts for those cases conforming with International Classification of Diseases code (ICD) 357.0, from January 1988 to December 1998. Clinical data such as age, sex, duration of hospitalization, antecedent events, date of onset, initial symptoms, other neurological symptoms and signs, results of lumbar puncture and electrophysiology, respiratory function and mortality, were recorded. RESULTS: We identified 49 cases of typical GBS, with non-specific age distribution. The calibrated male/female ratio was 1.54. The mean age for disease onset was greater for male patients (M: 54.8 years vs. F: 39.5 years), with more cases noted in winter. Upper respiratory tract infection (URI, 53%) was the most common antecedent event, followed, in descending order, by gastrointestinal symptoms (10%), skin lesions (6%) and ear infections (2%). The mean interval between antecedent events and disease onset was 10 +/- 3.1 days. Cranial nerve abnormalities, autonomic dysfunction and respiratory failure were not uncommon. The mortality rate was 6% (3/49). The abnormal findings from motor nerve conduction studies included prolonged distal latencies, reduced conduction velocities, reduced compound muscle action potentials and prolonged F-wave latencies. CONCLUSIONS: GBS occurs at all ages, is more prevalent among males, and is related to the season and antecedent infections. Continuous monitoring of autonomic and respiratory function is recommended. The results of motor nerve conduction studies suggest that the nerves fibers are affected at random, with multifocal distribution of the demyelinating lesions. The precise nature of the ataxia was not determined. Further studies are required to distinguish the etiology and expression of ataxia for GBS patients. PMID- 12583520 TI - Rehabilitation of dental implants for the post-irradiated and marginally resected mandible in an oral cancer patient. AB - We describe a case with severely compromised edentulous ridge in the mandible, which previously received marginal resection and radiotherapy due to oral cancer at the mouth floor. Through careful evaluation, the patient had 30 dives of hyperbaric oxygen therapy (HBO) before implant surgery. The edentulous ridge was rehabilitated with 4 endosteal implant fixtures, and palatal mucosa grafting vestibulosulcoplasty. The case had been followed for 4 years with stability of bone and a satisfactory result of rehabilitation. PMID- 12583521 TI - Zafirlukast-induced acute hepatitis. AB - Zafirlukast, a competitive cysteinyl leukotriene receptor antagonist, is a new class of asthma medications. It has shown an adverse event profile similar to that of placebo. Herein, we present a 69-year-old female patient who suffered from general malaise, poor appetite, nausea and jaundice after 3 months of zafirlukast therapy for asthma. She had no past history of liver disease, nor history of alcoholism, herb medication, blood transfusion, acupuncture, tattoo or recent traveling history. Liver biochemistries revealed elevated serum alanine aminotransferase and aspartase aminotransferase levels up to 481 U/L and 212 U/L, respectively. Moreover, peak serum total bilirubin level was elevated to 34.8 mg/dL during admission. Serum viral hepatitis marker, antinuclear antibody, anti mitochondrial antibody and anti-smooth muscle antibody were all negative. Her general condition and liver biochemistries improved gradually after zafirlukast was discontinued. Roussel Uclaf causality assessment for adverse drug reaction confirmed the diagnosis of drug-induced liver injury. This case reminds us that zafirlukast is a potentially hepato-toxic drug. If clinical manifestations of hepatitis develop, patients should be managed cautiously and closely monitored for liver biochemistries. If drug-induced hepatitis is suspected, medication should be discontinued immediately to prevent further liver injury. PMID- 12583522 TI - High dose chemotherapy and allogenic peripheral blood stem cell transplantation for multiple myeloma evolving from intra-abdominal plasmacytoma. AB - Solitary plasmacytomas include extramedullary plasmacytomas and those found in the bone. Seventy percent of patients are male and the median age is 50-55 years, younger than that for plasma cell myeloma. Most solitary plasmacytomas of bone eventually evolve to plasma cell myeloma within 2-10 years, while the extramedullary ones do so infrequently. We present an unusual case of intra abdominal plasmacytoma in a young woman which was misdiagnosed and treated as T cell lymphoma initially. Typical manifestations of plasma cell myeloma appeared one year later. High dose chemotherapy followed by allogeneic peripheral stem cell blood transplantation (allo-PBSCT) was given. Relapse in skin occurred one year after allo-PBSCT, and was treated with wide excision and local irradiation. The patient was well and alive without evidence of disease 4 years after wide excision of the recurrence of chest wall solitary plasmacytoma and local radiotherapy. PMID- 12583523 TI - Richard J. Roberts--Nobel Laureate for discovery of split genes. PMID- 12583524 TI - Effectiveness of radiotherapy for osteosarcoma that responds to chemotherapy. PMID- 12583526 TI - Computed tomographic determinants of neurologic deterioration in patients with large middle cerebral artery infarctions. AB - OBJECTIVE: To identify specific radiographic features on computed tomographic (CT) imaging that can predict neurologic deterioration in patients with large middle cerebral artery (MCA) infarctions. PATIENTS AND METHODS: We performed a 10 year retrospective review from January 1, 1991, through December 31, 2001, of medical records and CT scans of patients with large MCA infarctions. Neurologic deterioration was defied as progressive drowsiness or signs of herniation. The CT scans were grouped into 3 periods according to time after ictus. Radiographic features reviewed included hyperdense middle cerebral artery sign (HMCAS), more than a 50% loss of MCA territory, sulcal effacement, loss of lentiform nucleus or insular ribbon, and septal and pineal shift. Demographic and radiographic variables were compared by using t tests and the Fisher exact test. Prognostic values were calculated for all significant radiographic variables. RESULTS: Thirty-four CT scans in 22 patients before neurologic deterioration were compared with 47 scans obtained in 14 patients without neurologic worsening. There were no demographic differences between groups. Initial analysis revealed that early (<12 hours) involvement of more than 50% of the MCA territory (P=.047; odds ratio [OR], 14.02; 95% confidence interval [CI], 1.04-189.42) and the HMCAS at any time (P<.001; OR, 21.6; 95% CI, 3.54-130.04) were independent predictors of neurologic deterioration. The positive predictive power for early involvement of more than 50% of the MCA and the HMCAS was 0.75 and 0.91, respectively. CONCLUSION: The HMCAS and early CT evidence of more than 50% MCA involvement are predictive of neurologic deterioration in patients with large MCA infarcts. PMID- 12583525 TI - Neoadjuvant chemotherapy and local radiotherapy for high-grade osteosarcoma of the extremities. AB - OBJECTIVE: To determine the effectiveness of radiation therapy for local control of nonmetastatic osteosarcoma of the extremities after induction chemotherapy. PATIENTS AND METHODS: Of 187 patients with nonmetastatic osteosarcoma of the extremities treated with induction chemotherapy since 1986, 31 refused surgery and underwent standard, fractionated external beam radiotherapy for local control. The median radiation dose to the limb was 60 Gy (range 40-68 Gy). Records were reviewed through April 2002, and outcomes including radiologic and biochemical response, local control, limb function, and survival were analyzed. The end points were local progression-free survival, metastases-free survival, and overall survival. RESULTS: Overall survival, local progression-free survival, and metastases-free survival at 5 years were a mean +/- SD of 61%+/-11%, 56%+/ 12%, and 62%+/-10%, respectively. The outcome correlated significantly with patients' imaging and biochemical response. In patients who had a pronounced response, overall survival and metastases-free survival at 5 years were 90%+/-9% and 91%+/-9%, respectively, but it was only 35%+/-15% and 42%+/-13% in the nonresponders (P=.005 and P=.005, respectively). Local control was also related to response after induction chemotherapy. None of the 11 patients with both a good imaging and a good biochemical response had local relapse; median follow-up was 67 months. The estimated local progression-free survival among nonresponders was 31%+/-16% at 3 years and 0% at 5 years. Of 22 patients surviving without local disease progression, 19 (86%) had excellent limb function (Enneking score between 90% and 100%) at the time of most recent evaluation. CONCLUSION: When used after effective induction chemotherapy for osteosarcoma of the extremities, radiation therapy can be a reliable modality to control local disease and preserve limb function. PMID- 12583527 TI - Dietary intake of fruits, vegetables, and fat in Olmsted County, Minnesota. AB - OBJECTIVE: To assess self-reported dietary intake in the adult population of Olmsted County, Minnesota. SUBJECTS AND METHODS: We conducted a random-digit-dial telephone survey between March 1 and April 21, 1999, of 1232 adults residing in Olmsted County, Minnesota. We then mailed a structured questionnaire to the survey respondents and achieved a response rate of 732 individuals. Percentages of individuals and predictors of those who meet recommendations for intake of fruits and vegetables and for dietary fats were determined by using chi2 tests of general association and multivariate logistic regression. RESULTS: Only 16% of the population of Olmsted County reported meeting standard dietary recommendations for consuming both 5 or more servings of fruits and/or vegetables per day and no more than 30% of calories from fat. Fifty-one percent of the population was meeting neither recommendation. Women were more likely than men to report meeting both goals (22% vs 8%, P<.001), but still more women were meeting neither goal than were meeting both goals (40% vs 22%, P<.001). Multivariate logistic regression revealed the following factors to predict adherence to both goals: female sex, lower body mass index, nonsmoker, history of high cholesterol, and daily physical activity. CONCLUSION: Few individuals in Olmsted County are meeting national recommendations for intake of fruits, vegetables, and dietary fat. More effective interventions are needed to improve dietary habits in all subgroups of this community. PMID- 12583528 TI - Sympathotomy instead of sympathectomy for palmar hyperhidrosis: minimizing postoperative compensatory hyperhidrosis. AB - OBJECTIVE: To determine whether anatomical modification of sympathectomy diminishes severe postoperative compensatory hyperhidrosis. PATIENTS AND METHODS: From January 1, 2000, to June 1, 2001, we prospectively studied 10 consecutive patients with primary palmar hyperhidrosis, aged 14 to 42 years. Medical therapy had failed in all patients. A preoperative and postoperative evaluation and thermoregulatory sweat testing were conducted in all patients. All patients underwent sympathotomy (bilateral simple disconnection) of the second thoracic ganglion input into the brachial plexus, and no sympathetic ganglia were violated. RESULTS: In the 10 patients, all 20 upper extremities improved postoperatively: 11 (55%) had near-complete cessation of palmar sweating, 8 (40%) had marked reduction in sweating, and 1 (5%) had delayed onset but full sweating at the end of the thermoregulatory sweat test. No intraoperative complications of hemopneumothorax or Horner syndrome occurred. Importantly, there were no moderate or severe postoperative hyperhidrosis complications. CONCLUSION: Sympathotomy to disconnect T2 ganglion input into the brachial plexus produces excellent results in the treatment of primary palmar hyperhidrosis and may lower the severity of postoperative compensatory hyperhidrosis complications. Long-term durability of the procedure requires further follow-up. PMID- 12583529 TI - Polycythemia vera: a comprehensive review and clinical recommendations. AB - More than a century has elapsed since the appearance of the modern descriptions of polycythemia vera (PV). During this time, much has been learned regarding disease pathogenesis and PV-associated molecular aberrations. New information has allowed amendments to traditional diagnostic criteria. Phlebotomy remains the cornerstone treatment of PV, whereas myelosuppressive agents may augment the benefit of using phlebotomy for thrombosis prevention in high-risk patients. Excessive aspirin use is contraindicated in PV, although the use of lower-dose aspirin has been shown to be safe and effective in alleviating microvascular symptoms including erythromelalgia and headaches. Recent studies have shown the utility of selective serotonin receptor antagonists for treating PV-associated pruritus. Nevertheless, many questions remain unanswered. What is the specific genetic mutation or altered molecular pathway that is causally related to the disease? In the absence of a specific molecular marker, how is a working diagnosis of PV made? What evidence supports current practice in the management of PV? This article summarizes both old and new information on PV; proposes a modern diagnostic algorithm to formulate a working diagnosis; and provides recommendations for patient management, relying whenever possible on an evidence based approach. PMID- 12583530 TI - Recent advances in liver transplantation. AB - Advances in liver transplantation continue to evolve but are hampered by continued increasing shortages in donor organs. This has resulted in a high incidence of patients dying while on the United Network for Organ Sharing waiting list. Indeed, we continue to assess ways of expanding the donor pool by using marginal donors, living donor liver transplantation, split liver transplantation, domino transplantation, and hepatic support systems to prolong survival long enough for the patient to undergo liver transplantation. Changes in the liver allocation policy to reduce the number of people dying while waiting for an organ are discussed. Implementation of the model for end-stage liver disease allocation system should help alleviate the problem of increasing deaths of patients while on the waiting list. Recurrent disease, particularly recurrent hepatitis C, continues to be a major problem, and effective therapy is needed to prevent both progression of hepatitis C and recurrence in the graft and avoid retransplantation. The use of pegylated interferon in combination with ribavirin holds promise for improving the success in overcoming recurrent hepatitis C. Finally, advances in immunosuppression have reduced the incidence of acute cellular rejection and chronic rejection. However, these therapies have been fraught with metabolic complications that are now affecting quality of life and long-term survival. Tailoring immunosuppressive regimens to the individual patient is discussed. PMID- 12583531 TI - Two words to improve physician-patient communication: what else? AB - The medical interview is the physician's initial and perhaps most important diagnostic procedure, but physicians vary in their abilities and skills in physician-patient communication. Information gathering, relationship building, and patient education are the 3 essential functions of the medical interview. A physician-centered interview using a biomedical model can impede disclosure of problems and concerns. A patient-centered approach can facilitate patient disclosure of problems and enhance physician-patient communication. This, in turn, can improve health outcomes, patient compliance, and patient satisfaction and may decrease malpractice claims. Physicians can improve their communication skills through continuing education and practice. PMID- 12583532 TI - 67-year-old man with hepatitis C infection and abdominal distention. PMID- 12583533 TI - The kiss of death: a severe allergic reaction to a shellfish induced by a good night kiss. AB - True (IgE-mediated) allergic reactions to crustaceans are relatively common and can be serious. A young woman with a clinical history of multiple urticarial reactions after touching shrimp and lobster dishes was seen in an emergency department of a Mayo Clinic-affiliated hospital for an anaphylactic reaction that developed immediately after she kissed her boyfriend. The boyfriend had eaten several shrimp just before the kiss. It is important to warn susceptible patients that food does not actually have to be eaten to trigger an allergic reaction; touching the offending food and kissing or touching someone who has recently eaten the food can be enough to cause a major reaction. PMID- 12583534 TI - Fatal subacute cytomegalovirus encephalitis associated with hypogammaglobulinemia and thymoma. AB - Parathymic syndromes are systemic disorders that occur in association with thymoma. One such parathymic syndrome, hypogammaglobulinemia, was initially identified by Good in 1954 and has been referred to as Good syndrome. Patients with this syndrome develop a variety of recurrent infections due to the associated immunodeficiency. We describe a patient with cytomegalovirus encephalitis associated with Good syndrome and discuss the pathologic findings present on autopsy. The possibility of a cytomegalovirus infection should be considered early in the evaluation of patients with Good syndrome if appropriate clinical symptoms are present. PMID- 12583535 TI - Malignant hypertension as a presenting symptom of Takayasu arteritis. AB - We present an unusual case of malignant hypertension in a 20-year-old white woman. One week before hospitalization, she experienced occasional abdominal pain and claudication of both legs; otherwise, she had no remarkable medical history, including no history of high blood pressure. The origin of the patient's hypertension was renovascular, and the vascular injury was due to vasculitis of the large arteries. The combination of a difference in blood pressure between the patient's arms, angiographic findings, elevated erythrocyte sedimentation rate, and lack of markers for specific vasculitis led to the diagnosis of Takayasu arteritis. Surgical intervention was successful. PMID- 12583536 TI - Neurostimulation therapy for epilepsy: current modalities and future directions. AB - Neurostimulation is a recent development in the treatment of epilepsy. Vagus nerve stimulation (VNS), the only approved neurostimulation therapy for epilepsy to date, has proved to be a viable adjunctive treatment option. The exact mechanism of action of VNS is not fully understood. In 2 randomized double-blind trials, seizure frequency declined approximately 30% after 3 months of treatment. Long-term follow-up studies suggest that response improves over time, with approximately 35% of patients experiencing a 50% reduction and 20% experiencing a 75% reduction in seizure frequency after 18 months of treatment. Unfortunately, the number of patients rendered medication-free and seizure-free with VNS is low. Vagus nerve stimulation is best viewed as an option for patients who are not surgical candidates or who hesitate to take the risk of surgery yet continue to have seizures despite maximal medical therapy. Stimulation of other regions of the central nervous system for treating epilepsy, including the anterior and centromedian nuclei of the thalamus, the hippocampus, the subthalamic nucleus, and the cerebral neocortex, is currently under investigation. We review the history, proposed mechanisms of action, clinical trials, adverse effects, and future direction of VNS and other modalities of neurostimulation therapy for epilepsy. PMID- 12583537 TI - The anatomy laboratory: a concept ahead of its time. PMID- 12583538 TI - The red scalp sign. PMID- 12583539 TI - Invasive infections due to Apophysomyces elegans. PMID- 12583540 TI - Guidelines for performing single-platform absolute CD4+ T-cell determinations with CD45 gating for persons infected with human immunodeficiency virus. Centers for Disease Control and Prevention. AB - These guidelines were developed by CDC for laboratorians who perform immunophenotyping for detection and enumeration of CD4+ T-cells and other lymphocyte subsets in persons infected with human immunodeficiency virus (HIV). The guidelines describe single-platform technology (SPT), a process in which absolute counts of lymphocyte subsets are measured from a single tube by a single instrument. SPT incorporates internal calibrator beads of known quantity in the analysis of specimens by three- or four-color flow cytometry. With CD45 gating, the relative numbers of beads and lymphocyte subsets are enumerated, and their absolute numbers and percentage values are calculated. This report supplements previous recommendations published in 1997 (CDC. 1997 revised guidelines for performing CD4+ T-cell determinations in persons infected with human immunodeficiency virus [HIV]. MMWR 1997;46[No. RR-2]) that describe dual-platform technology, a method in which absolute counts are derived from measurements obtained from two instruments--a flow cytometer and hematology analyzer. The new recommendations address concerns specific to the implementation of SPT as well as other general topics such as laboratory safety and specimen handling. PMID- 12583541 TI - Guidelines for using the QuantiFERON-TB test for diagnosing latent Mycobacterium tuberculosis infection. Centers for Disease Control and Prevention. AB - Until 2001, the only test used to diagnose latent tuberculosis infection (LTBI) was the tuberculin skin test (TST). However, in 2001, a new test (QuantiFERON-TB or QFT; manufactured by Cellestis Limited, Carnegie, Victoria, Australia) that measures the release of interferon-gamma in whole blood in response to stimulation by purified protein derivative was approved by the Food and Drug Administration. This statement provides interim recommendations for using and interpreting QFT. As with TST, interpretation and indicated applications of QFT differ for persons according to their risk for LTBI and for developing tuberculosis (TB). This report provides guidance for public health officials, health-care providers, and laboratorians with responsibility for TB control activities in the United States in their efforts to incorporate QFT testing for detecting and treating LTBI. Regardless of the test used to identify LTBI, testing should be primarily targeted at diagnosing infected patients who will benefit from treatment. PMID- 12583542 TI - United States life tables, 2000. AB - This report presents period life tables for the United States based on age specific death rates in 2000. Data used to prepare these life tables are 2000 final mortality statistics; July 1, 2000, population estimates based on the 1990 decennial census; and data from the Medicare program. Presented are complete life tables by age, race, and sex. In 2000 the overall expectation of life at birth was 76.9 years, representing an increase of 0.2 years from life expectancy in 1999. Between 1999 and 2000, life expectancy increased for both males and females and for both the white and black populations. Life expectancy increased by 0.4 years for black males (from 67.8 to 68.2) and by 0.2 years for white males (from 74.6 to 74.8). It increased by 0.2 years for black females (from 74.7 to 74.9) and by 0.1 year for white females (from 79.9 to 80.0). PMID- 12583543 TI - Comparative in vitro activity of quinupristin/dalfopristin and seven other antimicrobials against methicillin-susceptible and methicillin-resistant nosocomial Staphylococcus aureus bloodstream isolates. AB - Staphylococcus aureus strains resistant to a variety of antimicrobial agents are often found in the hospital environment and are responsible for many life threatening infections. The activity of quinupristin/dalfopristin against 84 Staphylococcus aureus bloodstream isolates (both methicillin resistant and methicillin sensitive) was compared to the activity of vancomycin, teicoplanin, erythromycin, oxacillin, clindamycin, gentamicin, rifampicin. The Minimum Inhibitory Concentrations of these agents was evaluated with the Epsilometer Test. Quinupristin/dalfopristin inhibited all methicillin-sensitive strains at 1mg/L, and 75% of methicillin-resistant strains at 1.5mg/L. According to these results, quinupristin-dalfopristin shows promising in-vitro activity and may be a welcome alternative treatment for methicillin-resistant staphylococcal infections, resulting in reduced use of glycopeptides. PMID- 12583544 TI - A comparison between disk diffusion and microdilution for susceptibility testing of Mycobacterium fortuitum complex. AB - We evaluated a disk diffusion method using Mueller-Hinton agar for susceptibility testing of Mycobacterium fortuitum complex organisms. Ninety-five strains were tested both by broth microdilution and disk diffusion. Global results showed good correlation for all antimicrobials except for clarithromycin and erythromycin. However, when the results were analyzed according to species, correlation was poor except for a few antimicrobials. The analysis of the resistant/susceptible results was good for all the antimicrobials tested except azithromycin and erythromycin. In conclusion, the disk diffusion technique could be useful as a screening technique for some antibiotics, but the results must be confirmed by using an accepted reference technique. PMID- 12583545 TI - In vitro activity of thiamphenicol against multiresistant Streptococcus pneumoniae, Haemophilus influenzae and Staphylococcus aureus in Italy. AB - Thiamphenicol is a derivative of chloramphenicol characterized by a spectrum comparable to that of the parent compound against multiresistant pathogens but showing satisfactory tolerability. The in vitro activity of thiamphenicol and of 11 comparative drugs against 397 recently isolated antibiotic-resistant and/or invasive pneumococci and 52 multiply-resistant MRSA including 2 VISA strains was determined. Bactericidal activity against Haemophilus influenzae and the post antibiotic effect on Streptococcus pneumoniae, H. influenzae, Staphylococcus aureus and Escherichia coli were also assessed. Against invasive pneumococci, thiamphenicol and chloramphenicol were the most potent non-beta-lactam molecules together with vancomycin and rifampin. Against high-level penicillin-resistant strains phenicol activities were superior to those of cefotaxime, ceftriaxone and imipenem. Against MRSA thiamphenicol and chloramphenicol were second only to the glycopetides and also inhibited the VISA strains. Thiamphenicol showed a significant PAE (0.33 to 2.9h) on all pathogens studied and a powerful bactericidal effect against beta-lactamase-positive and -negative H. influenzae. These results indicate a good in vitro activity of thiamphenicol against difficult-to-treat multiply resistant pathogens. PMID- 12583546 TI - In-vitro activities of terbinafine, itraconazole and amphotericin B against Aspergillus and Cladosporium species. AB - The in vitro fungicidal and fungistatic activities of terbinafine were compared with those of itraconazole and amphotericin B against Aspergillus (n=63) and Cladosporium (n=21) isolates. The broth macrodilution modification of NCCLS reference method for filamentous fungi (M38-P) was used to assess the minimum inhibitory concentrations (MICs). Our data show that the in vitro activities of terbinafine were comparable to those of itraconazole and amphotericin B against the Aspergillus spp and Cladosporium strains tested. Despite strain-dependent variabilities, in general, itraconazole's activity was similar to that of amphotericin B against strains of Aspergillus and Cladosporium. Our data suggest that terbinafine may be useful in the treatment of Aspergillus and Cladosporium infections. PMID- 12583547 TI - A survey of antimicrobial drug resistance in respiratory tract pathogens, isolated in a northern Italian teaching hospital between 1990 and 1999. AB - Drug susceptibility test results of respiratory tract pathogens, isolated from patients admitted to the Clinic of Respiratory Diseases of the IRCCS San Matteo Hospital, University of Pavia (Italy) between 1990 and 1999, were retrospectively evaluated. A total of 1366 bacterial isolates were collected, including 499 gram positive and 867 gram-negative strains. In comparison to methicillin-susceptible Staphylococcus aureus, the methicillin-resistant strains (MRSA) showed high levels of resistance to many selected antibiotics, except for glycopeptides. Resistance rates to beta-lactams were high in both Pseudomonas aeruginosa and in the other gram-negative isolates, while aminoglycoside and ciprofloxacin resistance was less than 20%. Some pathogens became more resistant to selected antimicrobials during the observation period, including staphylococci to methicillin, MRSA to ciprofloxacin, P. aeruginosa isolates to imipenem and ciprofloxacin, and the other gram-negative strains to almost all drugs considered, with the exception of cefotaxime and cotrimoxazole. PMID- 12583548 TI - Effect of aminoglycosides (sub-MICs) on adherence of coagulase-negative Staphylococci to intraocular lens surface. AB - The authors evaluated the in vitro effects of subinhibitory concentrations (sub MICs) of aminoglycosides (gentamicin, netilmicin and tobramycin) on the adherence of coagulase-negative staphylococcus to intraocular lens (PMMA, acrylic and heparin) surface. The five strains of coagulase-negative Staphylococcus epidermidis used (S. epidermidis IMe7, IMe11, IMe27, IMe29 and IMe31) were tested for their ability to produce slime. The minimum inhibitory concentrations (MIC) indicated high susceptibility in vitro to the antibiotics and homogeneous values. The adhesion tests without antibiotics (K1, K2 and K3) for every time considered (30 min, 60 min and 180 min) and for all examined strains showed more constant adhesive activity on the surface of acrylic intraocular lens (IOLs) than on the surface of heparin IOLs. The aminoglycosides tested were able to reduce adhesivity by coagulase-negative S. epidermidis slime to the IOL surfaces, even at sub-MICs. The documentation obtained by scanning electron microscope shows the growth of a strain of S. epidermidis IMe11 after 180 minutes of contact which remains adherent to the surface of the acrylic IOLs. PMID- 12583549 TI - The integration of four major determinants of antibiotic action: bactericidal activity, postantibiotic effect, susceptibility, and pharmacokinetics. AB - A functional pharmacokinetic/pharmacodynamic (PK/PD) index that could simultaneously describe three controlling PD variables, i.e., bactericidal activity, postantibiotic effect (PAE), and susceptibility, in relation to pharmacokinetics, was designed using an in vitro kinetic model. Tobramycin was tested against one standard and five clinical strains of Pseudomonas aeruginosa. The organisms showed minimum inhibitory concentrations (MICs) ranging between 1 and >1000 microg/ml. The model allowed antibiotic concentrations to be reduced exponentially from initial concentrations at fixed multiples of MIC. Antibiotic removal was performed when the decreasing concentrations hit the MIC of individual strain to provide a wide range of AUC(>MIC) within an identical frame of AUC(>MIC)/MIC (AUIC) values. Viable counts were measured at antibiotic addition and before/after its removal for bactericidal activity and PAE assessments. A linear relationship was observed between PAE and bactericidal rate constants, though the pattern varied among different strains. Characterization of the exposure (AUC(>MIC))-effect relationships using the Emax model revealed that the less susceptible strains displayed lower Emax and higher EC50 for both antimicrobial effects. By employing the AUIC as a common frame of reference, regression analysis showed a significant linear correlation (p < 0.05) between the mean PAE and bactericidal rate data and, thereby simultaneously defining the four contributing factors of the PK/PD system. It appears that the AUIC, by conveying the pharmacokinetic and susceptibility information, could serve as a PK/PD index in bridging the interdependency of PAE and bactericidal activity. More importantly, the collective assessment of these four factors would allow more optimal evaluation of dosage regimens. PMID- 12583550 TI - In vivo pharmacodynamic evaluation of clarithromycin in comparison to erythromycin. AB - The efficacy of various dosing regimens of clarithromycin and erythromycin against recently isolated Streptococcus pneumoniae strains was determined in vivo using two animal infection models (mouse peritonitis and thigh infection). For the thigh infection model, mice received a total dose of 4 mg/Kg of either clarithromycin or erythromycin, as a single total dose or divided into 2, 4 or 8 doses/24h. After 24h of therapy S. pneumoniae organisms were killed at 2.06 to 4.03 log10 CFU/thigh by clarithromycin and the one- or two-dose regimens were significantly more effective than the four- or eight-dose regimens. Organism killing following 24h of therapy with erythromycin ranged from 1.13 to 2.31 log10 CFU/thigh, with the one- or two-dose regimens significantly less effective than the four- or eight-dose regimens. In the mouse survival study, the same dose of either clarithromycin or erythromycin was given as a single total dose or divided into two or four doses with dosing intervals of 4 and 2-times the t1/2 respectively. The results obtained in this model show that there is a significant difference in survival when clarithromycin is administered less frequently (4% deaths for the one-dose regimen in comparison to 40% deaths with the four-dose regimen, P < 0.01, Chi-square test). With erythromycin there was a trend for increased survival with the multiple-dose regimen, with significantly higher survival when concentrations exceeding the MIC were maintained for a longer time period. These results indicate that the time during which serum concentrations exceeding the MIC value of the pathogen is an important parameter for efficacy for erythromycin. On the contrary, results with both animal models demonstrate that bacterial killing and survival are significantly higher among clarithromycin treated mice when the antibiotic is administered less frequently and the highest Cmax/MIC ratio is achieved. PMID- 12583551 TI - Efficacy of amoxicillin-sulbactam, given twice-a-day, for the treatment of community-acquired pneumonia: a clinical trial based on a pharmacodynamic model. AB - The present multicenter study reports the results of a clinical trial, designed on the basis of a pharmacodynamic study published previously (Bantar et al., J. Chemother 2000; 12: 223-227) to assess the efficacy of amoxicillin/sulbactam (875 mg/125 mg), given orally twice-a-day for 7 days in the treatment of patients with community-acquired pneumonia (CAP). Eighty-four evaluable subjects older than 19 years with clinical symptoms and features suggestive of CAP, consulting from June 2000 to March 2002 and meeting the PORT risk class I through III, were enrolled in the study. Mean age (y +/- standard deviation) was 46.7 +/- 16.3 and 62% of the patients had some co-morbidity predisposing for CAP. Several individuals (77.4%) fell into a low-risk class (i.e. PORT I or II) and 22.6% of patients belonged to a moderate-risk class at the start of treatment. Six patients (6.45%) had pneumococcal bacteremia. Streptococcus pneumoniae was the organism most frequently isolated (61.9% of all the patients in whom an etiologic diagnosis was made), followed by Haemophilus influenzae. Clinical success was observed in 97.6% of the patients (confidence interval 95%, 94.3%-100%). Almost all the individuals with clinical success became afebrile within the first 3 days of therapy. Ten patients (11.8%) reported mild or moderate adverse events (especially diarrhea) possibly related to the antimicrobial therapy, but this did not lead to withdrawal from the trial. The results of this study suggest that amoxicillin/sulbactam (875 mg/125 mg) is an efficacious and well tolerated option for treating patients with CAP belonging to a low-moderate risk class and support the use of a short, oral (7-day) b.i.d. regimen. PMID- 12583552 TI - Efficacy and safety of short course (5-day) moxifloxacin vs 7-day ceftriaxone in the treatment of acute exacerbations of chronic bronchitis (AECB). AB - The aim of this multicenter, open, randomized study was to compare the efficacy and tolerability of a 5-day treatment course with oral moxifloxacin (MXF) vs a 7 day course with i.m. ceftriaxone (CRO) in 476 patients with acute exacerbations of chronic bronchitis (AECB), and to conduct a cost minimization analysis of the two treatments from the perspectives of both the Italian National Health Service (INHS) and society. The study was conducted in Italy. Clinical success rates at test-of-cure in the 423 patients of the PP (Per Protocol) population (primary efficacy parameter) were 90.6% and 89.0% for MXF and CRO, respectively. Statistical non-inferiority of MXF vs CRO was confirmed. Similar results were found between study drugs on the secondary efficacy parameters, including success at end-of-treatment (95.3% for MXF vs 92.9% for CRO), success at test-of-cure in bacteriologically-positive patients (94.1% vs 90.7%) and eradication/presumed eradication rates (91.7% vs 93.3%). ITT (Intention-to-Treat) analysis confirmed these data. There was a low incidence of adverse events (10.8% vs 9.1%). During a 6-month follow-up period, relapse rates were lower for MXF vs CRO (23.3% vs 28.3%; p > .05). Compared with CRO, MXF was associated with cost savings per patient ranging from Euro226.57 (INHS perspective) to Euro448.23 (societal perspective), with lower hospitalization rate the major variable contributing to reduced costs. MXF appears to be an ideal candidate for AECB treatment. PMID- 12583553 TI - Carbapenems in the treatment of severe community-acquired pneumonia in hospitalized elderly patients: a comparative study against standard therapy. AB - In this open, prospective, study were enrolled 204 hospitalized elderly patients with severe (88 males, 116 females, age range 70-94). Patients were randomized to receive one of the following antibiotic treatment regimens: meropenem 500 mg i.v. t.i.d. (52); imipenem/cilastatin 500 mg i.v. t.i.d. (51), clarithromycin 500 mg + ceftriaxone 1 g i.v. b.i.d. (52), clarithromycin 500 mg + amikacin 250 mg i.v. b.i.d. (49). In 99 cases causative germs were isolated (24 meropenem, 26 imipenem, 23 clarithromycin + ceftriaxone, 26 ceftriaxone + amikacin). A satisfactory clinical, bacteriological response was achieved respectively in 86.5% 77% in meropenem; 86.3% 71% in imipenem/cilastatin; 69% 61% in ceftriaxone + clarithromycin and in 85.7% 77% in clarithromycin + amikacin. The mean total cost for each patient was $1,560; $1,620; $1,760 and $1,792 in meropenem, imipenem/cilastatin, clarithromycin + ceftriaxone and clarithromycin + amikacin respectively. This study shows that treatment with either meropenem or imipenem is as efficacious as conventional therapy in the treatment of community acquired pneumonia (CAP), and that meropenem is the most cost-effective. PMID- 12583554 TI - Antimicrobial prophylaxis in laparotomic gynecologic surgery: a prospective randomized study comparing amoxicillin-clavulanic acid with cefazolin. AB - A prospective randomized study was conducted at the Department of Obstetrics and Gynecology, University of Bari to compare two antimicrobial regimens, amoxicillin clavulanic acid with cefazolin as ultra-short term prophylaxis in laparotomic gynecologic surgery. Patients were randomly allocated to receive a single dose of amoxicillin-clavulanic acid (2.2 g) [Group A] or cefazolin (2 g) [Group B] 30 minutes before surgery. Each patient was assessed daily until discharge for fever and the presence of infection of the surgical wound, urinary tract and respiratory tract. In the amoxicillin-clavulanic acid (Group A) and cefazolin (Group B) groups, overall 258 and 253 patients, respectively were evaluable for prophylactic efficacy at hospital discharge. Infectious complications were infrequent in both arms. Febrile morbidity occurred in 16 (6.3%) and 21 (8.1%) patients respectively in the amoxicillin-clavulanic acid and cefazolin groups. Wound infection and urinary tract infection were also higher but not significantly in the cefazolin group (0.8% versus 0% and 2.7% versus 2.0% respectively). There was no respiratory tract infection or septic death in either group. It is concluded that ultra-short term prophylaxis with both amoxicillin clavulanic acid and cefazolin is safe and effective in elective laparotomic gynecologic surgery. PMID- 12583555 TI - Etoposide, doxorubicin and cisplatin alternating with 5-fluorouracil, doxorubicin and high-dose methotrexate in patients with advanced adenocarcinoma of the stomach or the gastroesophageal junction. AB - Both the etoposide, doxorubicin, cisplatin (EAP) and 5-fluorouracil, doxorubicin, high-dose methotrexate (FAMTX) schedules have been reported to be active in advanced gastric cancer. Since these regimens include non-cross resistant agents, a regimen that consists of EAP alternating with FAMTX may have an advantage over each regimen alone. We undertook a phase II trial to evaluate EAP/FAMTX in patients with advanced adenocarcinoma of the stomach or gastroesophageal junction. Of the 56 patients treated, an objective response was observed in 34%, including complete response in 7%. Median response duration was 8 months and median survival for the entire group was 9 months. The main toxicity was myelosuppression. Hospitalization for granulocytopenic fever was required in 32% of patients and 34% required red blood cells (RBC) transfusion. Non-hematological toxicity was moderate. There were three drug-related deaths associated with granulocytopenic fever. We conclude that the alternating EAP/FAMTX regimen is associated with occasional lethal events and has no obvious advantage over either regimen alone. PMID- 12583556 TI - Successful treatment of Candida albicans osteomyelitis of the spine with fluconazole and surgical debridement: case report. AB - A 65-year old diabetic male presented with progressive bone destruction of thoracic spine (T-11&12) with cord compression. Candida albicans was isolated from aspirated materials pre-and intra-operative. Two weeks of fluconazole was given prior to surgical debridement, and fixation of the lesion. C. albicans isolated pre-and 2-weeks after fluconazole treatment were DNA-typed using AP-PCR. MIC was 2-4 mg/l in all isolates tested. The pre-and post treatment isolates had two DNA patterns, indicating the existence of two different strains. Surgical treatment was necessary for patient recovery. PMID- 12583557 TI - Intramedullary spinal cord metastases in breast carcinoma: report of two cases and review of the literature. AB - Intramedullary spinal cord metastases (ISCM) are usually the result of rapidly progressing systemic malignancy. Breast cancer is one of the most common solid tumors with a high propensity of CNS dissemination. In the present report we describe two new cases with advanced breast cancer developing ISCM after a variable disease course. One of these patients had brain metastases at presentation, while at relapse developed leptomeningeal carcinomatosis which was treated successfully, but followed shortly, as a terminal event, by ISCM and parenchymal brain recurrence. The other patient was treated initially for locally advanced breast cancer and after multiple locoregional relapses, she developed liver metastases and subsequent ISCM and asymptomatic parenchymal brain deposits. Both patients experienced a rather rapidly evolving disease course leading to death 2 and 4 months, respectively, after widespread neuraxis dissemination of their cancer. Both these cases, added to the list of the anecdotally reported cases of ISCM after breast cancer, undermine the ominous prognosis and limited treatment options available for this disease manifestation, and an extensive literature review and discussion of similar cases is provided. PMID- 12583558 TI - Acute community-acquired pneumonia in adults: guidelines for initial antimicrobial therapy based on local evidence from a South American working group (ConsenSur). PMID- 12583559 TI - Multiresistant Escherichia coli isolated from women with community-acquired urinary tract infections in the Gaza Strip. PMID- 12583560 TI - Nosocomial Stenotrophomonas maltophilia strain with a broad host range ability to transfer antibiotic resistance. PMID- 12583561 TI - Prevalence and antifungal susceptibilities of yeast isolates during one year at a university hospital in Turkey. PMID- 12583562 TI - Is academic radiology still attractive? PMID- 12583563 TI - Influence of gravity on radiographic contrast material-based measurements of regional blood flow distribution. AB - RATIONALE AND OBJECTIVES: Radiographic measurement of regional blood flow distribution in the lungs is potentially biased because the contrast material used to track flow is denser than blood. The authors performed this study to evaluate the effect of gravity on flow estimates by using an experimental test phantom and numeric simulations. MATERIALS AND METHODS: Cross-sectionally uniform boluses of radiopaque contrast material were delivered at the upstream end of a horizontal inlet tube connected to a downstream axisymmetric bifuration attached to collecting tubing spirals. The phantom was imaged by using both planar angiography and dynamic multi-detector row computed tomography (CT) during the passage of the bolus through the phantom. The images were analyzed to determine the relative amounts of contrast material traveling through the top and bottom branches of the bifurcation by using varying Reynolds numbers and ratios of inlet tube volume to bolus volume. Numeric simulations of flow within a straight channeL with use of a dispersion operator intended to simulate settling of the bolus due to gravity, were performed under conditions representative of those in the experiments. RESULTS: When the plane of the bifurcation was vertical and actual flow through the two branches was equal, the fraction of contrast material passing through the downward-directed branch increased with decreasing Reynolds number and increasing inlet tube-bolus volume ratio. This occurred in both the experiments and the simulations. CONCLUSION: Because in the circulation Reynolds number decreases and pathway length increases with decreasing vessel diameter, the accuracy of regional flow measurements obtained with angiography or CT within the lungs may be limited by density differences between contrast material and blood. PMID- 12583564 TI - Vesicoureteral reflux imaging in children: comparative cost analysis. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to compare the costs of voiding cystourethrography (VCUG) versus radionuclide cystography (RNC) for evaluation of vesicoureteral reflux in children. MATERIALS AND METHODS: The variable direct costs of performing 25 VCUG and 25 RNC examinations in age- and general health-matched patients suspected of having vesicoureteral reflux was determined by using time and motion analyses. All personnel directly involved in the cases were tracked, and the involvement times were recorded to the nearest minute. All material items used during the procedures were recorded. The cost of labor was determined from personnel reimbursement data, and the cost of materials, from vendor pricing. The fixed direct costs were assessed from hospital accounting records. Mean, standard deviation, and 95% confidence interval (CI) were determined for all direct (fixed and variable) costs. The total costs were determined for each procedure and compared by using the Student t test. RESULTS: There was a significant difference (P < .0001) between the mean total direct cost of VCUG ($112.17 +/- 10.33) and that of RNC ($64.58 +/- 1.91). VCUG examination for vesicoureteral reflux in children cost 1.74 times more than RNC examination (95% CI: 1.28, 2.36). CONCLUSION: When the technique is clinically appropriate, institutions may obtain substantial cost savings by using RNC in place of VCUG for examining children suspected of having vesicoureteral reflux. PMID- 12583565 TI - Hydrogen-1 MR spectroscopy of the peritumoral zone in patients with cerebral glioma: assessment of the value of the method. AB - RATIONALE AND OBJECTIVES: The determination of tumor boundaries, especially in high-grade glioma, is critically important for the proper planning of treatment, but the standard diagnostic imaging methods do not enable precise delimitation of the extent of tumor cell infiltration into the surrounding tissue. The objective of this study was to assess the usefulness of hydrogen-1 (H-1) magnetic resonance (MR) spectroscopy for determining the extent of gliomatous infiltrate in the "uncertain zone'--the peritumoral area that appears unchanged on standard diagnostic MR images. MATERIALS AND METHODS: The study group consisted of 64 patients with cerebral glioma scheduled for tumor resection and subsequent radiation therapy. All patients were examined prior to resection with MR imaging and MR spectroscopy. MR spectra were obtained from examination of the solid part of the tumor and from two peritumoral volumes of interest located approximately along the axis of surgical access to the tumor. MR spectra obtained from a group of 32 healthy volunteers were used as control data. RESULTS: Analysis of the consequent voxels in the peritumoral zone revealed statistically significant differences in lipid/creatine and lactate/creatine metabolite ratios between patient subgroups with recurrent malignant lesions and without recurrent lesions. Significant differences also were found between the patient group and the control group in most metabolite ratios assessed. CONCLUSION: H-1 MR spectroscopic demonstration of metabolic changes in the peritumoral zone can guide treatment for cerebral glioma, enabling the physician to identify patients who have a high risk of recurrence. PMID- 12583566 TI - Multiple neural network classification scheme for detection of colonic polyps in CT colonography data sets. AB - RATIONALE AND OBJECTIVES: A new classification system for colonic polyp detection, designed to increase sensitivity and reduce the number of false positive findings with computed tomographic colonography, was developed and tested in this study. MATERIALS AND METHODS: The system involves classification by a committee of neural networks (NNs), each using largely distinct subsets of features selected from a general set. Back-propagation NNs trained with the Levenberg-Marquardt algorithm were used as primary classifiers (committee members). The set of features included region density, Gaussian and mean curvature and sphericity, lesion size, colon wall thickness, and the means and standard deviations of all of these values. Subsets of variables were initially selected because of their effectiveness according to training and test sample misclassification rates. The final decision for each case is based on the majority vote across the networks and reflects the weighted votes of all networks. The authors also introduce a smoothed cross-validation method designed to improve estimation of the true misclassification rates by reducing bias and variance. RESULTS: This committee method reduced the false-positive rate by 36%, a clinically meaningful reduction, and improved sensitivity by an average of 6.9% compared with decisions made by any single NN. The overall sensitivity and specificity were 82.9% and 95.3%, respectively, when sensitivity was estimated by means of smoothed cross-validation. CONCLUSION: The proposed method of using multiple classifiers and majority voting is recommended for classification tasks with large sets of input features, particularly when selected feature subsets may not be equally effective and do not provide satisfactory true- and false-positive rates. This approach reduces variance in estimates of misclassification rates. PMID- 12583567 TI - Using a human visual system model to optimize soft-copy mammography display: influence of display phosphor. AB - RATIONALE AND OBJECTIVES: The authors developed an efficient method for optimizing cathode ray tube performance for soft-copy digital mammography displays, based on correlation between the performance of human observers and the performance of a mathematical computer model of the human visual system. The authors measured radiologist performance on soft-copy display monitors with different phosphors and used these results to validate the human visual performance model. MATERIALS AND METHODS: Six radiologists viewed a series of 250 mammographic images with microcalcifications of different contrast levels. They viewed images on two soft-copy display monitors with phosphor luminescence-one with P45 and the other with P104. The same images were analyzed with the JNDmetrix Visual Discrimination Model, which is based on psychophysical just noticeable difference measurement principles and on frequency-channel vision modeling principles. Receiver operating characteristic curves were generated for the human and model observers' performances, and results were compared statistically. RESULTS: Both human and model performance (area under the receiver operating characteristic curve) was better overall with the P45 than with the P104 monitor, especially for microcalcifications in the midlevel contrast range. There was high correlation between the human and model observers. CONCLUSION: The results indicate that the type of phosphor in a display monitor can influence observer performance significantly and that a model based on characteristics of the human visual system can be used to predict human observer performance accurately. PMID- 12583568 TI - Utility of high-resolution CT for management of diffuse lung disease: results of a survey of U.S. pulmonary physicians. AB - RATIONALE AND OBJECTIVES: This study was performed to determine how U.S. pulmonologists rate the clinical contributions of high-resolution computed tomography (CT) in patients with diffuse lung disease, to ascertain how the technique affects management decisions, and to determine the effect of three physician characteristics on these attitudes. MATERIALS AND METHODS: The authors surveyed 450 practicing pulmonologists. The questionnaire explored perceptions of the efficacy of high-resolution CT for achieving five clinical objectives, the importance of high-resolution CT in 17 diseases, and the effects of the CT results on management decisions. Responses were examined by type of clinical practice, monthly referral volume, and year of completion of pulmonary fellowship. RESULTS: The response rate was 52.6%. High-resolution CT was rated most helpful for determining the extent of diffuse lung disease and least helpful for assessing disease activity and prognosis. Pulmonologists believed that high resolution CT was most important in the idiopathic interstitial pneumonias and least important in Pneumocystis carinii pneumonia and emphysema. High-resolution CT results frequently increased the physician's confidence in a presumptive diagnosis. Recently trained pulmonologists were more likely to report that high resolution CT results altered their management plan. There were no significant differences related to type of practice. CONCLUSION: U.S. pulmonologists in a variety of practice settings value the contributions of high-resolution CT in patients with diffuse lung disease and find it particularly important in the chronic interstitial pneumonias. High-resolution CT results have their greatest effect in confirming a presumptive clinical diagnosis and less frequently alter the management plan. PMID- 12583569 TI - The concept of image-guided therapy. AB - Parallel with current applications in minimally invasive surgery, the introduction of new imaging modalities, and the availability of high-performance computing, new image-guided therapies are being developed at an impressive rate. Indeed, across a broad front of imaging technologies, rapid advances are being realized. Vastly refined technology for processing and using images, as well as improved therapeutic end-effectors, have no doubt hastened this remarkable progress. At the same time, advances in clinical evaluation and complementary technologies will provide the necessary infrastructure through which IGT can be applied in diverse therapeutic settings--from the already well-established neurosurgical applications to the thermal ablation of tumors in organs other than the brain. That IGT is more efficient and effective and less expensive than conventional surgery has been confirmed both in extensive, long-term studies and in ongoing, revolutionary applications in the operating room. We have laid critical groundwork with this extraordinary technology and have now begun to realize quantifiable benefits in terms of improved surgical and patient outcomes. PMID- 12583571 TI - Support vector machines for diagnosis of breast tumors on US images. AB - RATIONALE AND OBJECTIVES: Breast cancer has become the leading cause of cancer deaths among women in developed countries. To decrease the related mortality, disease must be treated as early as possible, but it is hard to detect and diagnose tumors at an early stage. A well-designed computer-aided diagnostic system can help physicians avoid misdiagnosis and avoid unnecessary biopsy without missing cancers. In this study, the authors tested one such system to determine its effectiveness. MATERIALS AND METHODS: Many computer-aided diagnostic systems for ultrasonography are based on the neural network model and classify breast tumors according to texture features. The authors tested a refinement of this model, an advanced support vector machine (SVM), in 250 cases of pathologically proved breast tumors (140 benign and 110 malignant), and compared its performance with that of a multilayer propagation neural network. RESULTS: The accuracy of the SVM for classifying malignancies was 85.6% (214 of 250); the sensitivity, 95.45% (105 of 110); the specificity, 77.86% (109 of 140); the positive predictive value, 77.21% (105 of 136); and the negative predictive value, 95.61% (109 of 114). CONCLUSION: The SVM proved helpful in the imaging diagnosis of breast cancer. The classification ability of the SVM is nearly equal to that of the neural network model, and the SVM has a much shorter training time (1 vs 189 seconds). Given the increasing size and complexity of data sets, the SVM is therefore preferable for computer-aided diagnosis. PMID- 12583570 TI - New assistive devices for MR-guided microwave thermocoagulation of liver tumors. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to develop and test new assistive devices to aid in the performance of safe, easy, and accurate microwave thermocoagulation therapy guided by magnetic resonance (MR) imaging. MATERIALS AND METHODS: An open-configuration MR imager with an optical tracking system for image plane control and a microwave coagulator operating at 2.45 GHz were used. Liver tumors were percutaneously punctured under the guidance of MR images and ablated. Handpiece adapters were then prepared for the optical tracking system. An MR-compatible endoscopic system also was used. Navigation software was installed and customized. These devices were combined to provide near-real-time MR image-guided navigation during thermocoagulation therapy for liver tumors. RESULTS: The handpiece adapters improved the flexibility of approaches for percutaneous puncture of tumors. The MR-compatible endoscope was used as a thoracoscope, and tumors just below the diaphragm were safely and easily punctured through the diaphragm. The customized navigation software displayed near-real-time MR images together with two reformatted images (on the same plane and on a perpendicular plane) from the registered high-resolution three dimensional data, enabling accurate MR imaging navigation. CONCLUSION: These new assistive devices made it easier to perform safe, easy, and accurate MR-guided thermocoagulation therapy of liver tumors. PMID- 12583572 TI - Classification of galactograms with ramification matrices: preliminary results. AB - RATIONALE AND OBJECTIVES: The poor specificity of galactography, the imaging modality generally indicated in cases of nipple discharge, has led to a large number of biopsies with negative results. A quantitative scheme for classifying galactographic findings might help reduce the number of such biopsies in the future. As a first step toward that goal, the authors have studied one quantitative method for describing the branching of ducts by using ramification matrices (R matrices), and the correlation of the values of the matrix elements with clinical findings. MATERIALS AND METHODS: The ductal trees were manually segmented for 25 galactographic views from 15 patients, and corresponding R matrices were calculated. Patients were divided into two groups: those with no reported galactographic findings (NF) and those with reported findings (RF) of ductal ectasia, cysts, or papilloma. In a leave-one-out fashion, the authors evaluated a classification scheme that was based on R-matrix coefficients and used a Bayesian decision rule. The effects of segmentation were tested by successively removing each of the terminal ducts and computing the corresponding matrices of the pruned trees. RESULTS: With use of a single R-matrix element, 92% and 62% of NF and RF cases were correctly classified, respectively (P = .007). With use of two elements, 83% and 77% of NF and RF cases were correctly classified, but this result was not statistically significant (P = .108). In a test of robustness, an analysis of pruned trees yielded an average root-mean square fractional difference of 9.7% between the elements of the original and the R matrix averaged over all pruned trees. CONCLUSION: The preliminary analysis indicates that it may be possible to identify cases with reported galactographic findings by using R matrices. PMID- 12583573 TI - Incorporating electronic media into medical student education: a survey of AMSER members on computer and web use in radiology courses. Alliance of Medical Student Educators in Radiology. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to define the current use of information technology in radiology tutorials for medical students. MATERIALS AND METHODS: The authors conducted a Web-based survey of directors of medical school courses in radiology. The survey dealt with the details of the courses and the use of computers and the Web during the courses. RESULTS: There were 48 responses. Most radiology courses were elective (73%) and were offered monthly. Most institutions (79%) had picture archiving and communication systems (PACS) available or were completely filmless. The teaching case presentations, however, often included film images displayed on a view box or by an overhead projector. Computers dedicated to student use were uncommon (28%). The Web was used infrequently as a teaching resource, and a Web site was not available in most courses. Computer technical support was variable and usually provided by the course director. Course directors at institutions with PACS were more likely to use digital technology for case presentations and more likely to use the Web for teaching purposes. CONCLUSION: Despite the widespread use of digital technology and PACS in the field of radiology, digital technology is underused in radiology courses. However, departments with PACS tend to use digital technology more frequently in education than do departments without PACS. PMID- 12583574 TI - Independent versus sequential reading in ROC studies of computer-assist modalities: analysis of components of variance. PMID- 12583575 TI - Perspective: purging the genetic load: a review of the experimental evidence. AB - Inbreeding depression, the reduction in fitness that accompanies inbreeding, is one of the most important topics of research in evolutionary and conservation genetics. In the recent literature, much attention has been paid to the possibility of purging the genetic load. If inbreeding depression is due to deleterious alleles, whose effect on fitness are negative when in a homozygous state, then successive generations of inbreeding may result in a rebound in fitness due to the selective decrease in frequency of deleterious alleles. Here we examine the experimental evidence for purging of the genetic load by collating empirical tests of rebounds in fitness-related traits with inbreeding in animals and plants. We gathered data from 28 studies including five mammal, three insect, one mollusc, and 13 plant species. We tested for purging by examining three measures of fitness-component variation with serial generations of inbreeding: (1) changes in inbreeding depression, (2) changes in fitness components of inbred lines relative to the original outbred line, and (3) purged population (outcrossed inbred lines) trait means as a function of ancestral outbred trait means. Frequent and substantial purging was found using all three measures, but was particularly pronounced when tracking changes in inbreeding depression. Despite this, we found little correspondence between the three measures of purging within individual studies, indicating that the manner in which a researcher chooses to estimate purging will affect interpretation of the results obtained. The discrepancy suggests an alternative hypothesis: rebounds in fitness with inbreeding may have resulted from adaptation to laboratory conditions and not to purging when using outcrossed inbred lines. However, the pronounced reduction in inbreeding depression for a number of studies provides evidence for purging, as the measure is likely less affected by selection for laboratory conditions. Unlike other taxon-specific reviews on this topic, our results provide support for the purging hypothesis, but firm predictions about the situations in which purging is likely or the magnitude of fitness rebound possible when populations are inbred remain difficult. Further research is required to resolve the discrepancy between the results obtained using different experimental approaches. PMID- 12583576 TI - Deleterious mutation and the evolution of eusociality. AB - Certain arguments concerning the evolution of eusociality form a classic example of the application of the principles of kin selection. These arguments center on the different degrees of relatedness of potential beneficiaries of an individual's efforts, for example a female's higher relatedness to her sisters than to her daughters in a haplodiploid system. This type of reasoning is insufficicnt to account for the evolution and maintainence of sexual reproduction, because parthenogenic females produce offspring that are more closely related to them than are offspring produced sexually. Among the forces invoked to explain sexual reproduction is deleterious mutation. This factor can be shown to favor eusociality as well, because siblings produced by helping carry fewer deleterious alleles on average than would offspring. The strength of this effect depends on the genomewide deleterious mutation rate, U, and on the selection coefficient, s, associated with deleterious alleles. For small s, the effect depends approximately on the product Us. This phenomenon illustrates that an assumption implicit in some analyses-that the relatedness of an individual to an actor is all that matters to its value to that actor-can fail for the evolution of eusociality as it does for the evolution of sex. PMID- 12583577 TI - A metapopulation perspective on genetic diversity and differentiation in partially self-fertilizing plants. AB - Partial self-fertilization is common in higher plants. Mating system variation is known to have important consequences for how genetic variation is distributed within and among populations. Selfing is known to reduce effective population size, and inbreeding species are therefore expected to have lower levels of genetic variation than comparable outcrossing taxa. However, several recent empirical studies have shown that reductions in genetic diversity within populations of inbreeding species are far greater than the expected reductions based on the reduced effective population size. Two different processes have been argued to cause these patterns, selective sweeps (or hitchhiking) and background selection. Both are expected to be most effective in reducing genetic variation in regions of low recombination rates. Selfing is known to reduce the effective recombination rate, and inbreeding taxa are thus thought to be particularly vulnerable to the effects of hitchhiking or background selection. Here I propose a third explanation for the lower-than-expected levels of genetic diversity within populations of selfing species; recurrent extinctions and recolonizations of local populations, also known as metapopulation dynamics. I show that selfing in a metapopulation setting can result in large reductions in genetic diversity within populations, far greater than expected based the lower effective population size inbreeding species is expected to have. The reason for this depends on an interaction between selfing and pollen migration. PMID- 12583578 TI - Invasion of unisexuals in hermaphrodite populations of animal-pollinated plants: effects of pollination ecology and floral size-number trade-offs. AB - The most common sexual system in animal-pollinated plants is hermaphroditism, while some species are dioecious or gynodioecious and a very few are androdioecious. In this paper, I attempt to explain this pattern by extending previous models for the evolution of sexual systems to incorporate two main features: (1) a portion of investment in pollinator attraction contributes to only female or male function, because one sexual function of a flower is saturated with pollinator visitation earlier than the other sexual function; and (2) there are trade-offs between the size and number of flowers. The analysis was conducted to determine the conditions when females and males can increase in frequency in a hermaphroditic population, assuming either concave or convex pollinator gain curves (relation between investment to attractive structures of a flower and frequency of pollinator visits to the flower). The results suggest that both of the main factors play important roles in the evolution of plant sexual systems: uneven contribution of pollinator-attractive structures and nonlinear trade-offs between flower size and number can destabilize hermaphroditism. When a convex pollinator gain curve was assumed, the effect of nonlinear trade-offs can produce accelerating compensation from the elimination of one sexual function, allowing males to increase for large regions of parameter space, where females could not increase. The last prediction obviously conflicts with the observed rarity of androdioecy in nature, indicating the necessity of exploring pollinator gain curves in more detail. PMID- 12583579 TI - Phylogeographic breaks without geographic barriers to gene flow. AB - The spatial distribution of genetic markers can be useful both in estimating patterns of gene flow and in reconstructing biogeographic history, particularly when gene genealogies can be estimated. Genealogies based on nonrecombining genetic units such as mitochondrial and chloroplast DNA often consist of geographically separated clades that come into contact in narrow regions. Such phylogeographic breaks are usually assumed to be the result of long-term barriers to gene flow. Here I show that deep phylogeographic breaks can form within a continuously distributed species even when there are no barriers to gene flow. The likelihood of observing phylogeographic breaks increases as the average individual dispersal distance and population size decrease. Those molecular markers that are most likely to show evidence of real geographic barriers are also most likely to show phylogeographic breaks that formed without any barrier to gene flow. These results might provide an explanation as to why some species, such as the greenish warblers (Phylloscopus trochiloides), have phylogeographic breaks in mitochondrial or chloroplast DNA that do not coincide with sudden changes in other traits. PMID- 12583580 TI - Phylogeny of Acridocarpus-Brachylophon (Malpighiaceae): implications for tertiary tropical floras and Afroasian biogeography. AB - A major tenet of African Tertiary biogeography posits that lowland rainforest dominated much of Africa in the late Cretaceous and was replaced by xeric vegetation as a response to continental uplift and consequent widespread aridification beginning in the late Paleogene. The aridification of Africa is thought to have been a major factor in the extinction of many African humid tropical lineages, and in the present-day disparity of species diversity between Africa and other tropical regions. This primarily geologically based model can be tested with independent phylogenetic evidence from widespread African plant groups containing both humid- and xeric-adapted species. We estimated the phylogeny and lineage divergence times within one such angiosperm group, the acridocarpoid clade (Malpighiaceae), with combined ITS, ndhF, and trnL-F data from 15 species that encompass the range of morphological and geographic variation within the group. Dispersal-vicariance analysis and divergence-time estimates suggest that the basal acridocarpoid divergence occurred between African and Southeast Asian lineages approximately 50 million years ago (mya), perhaps after a southward ancestral retreat from high-latitude tropical forests in response to intermittent Eocene cooling. Dispersion of Aeridocarpus from Africa to Madagascar is inferred between approximately 50 and 35 mya, when lowland humid tropical forest was nearly continuous between these landmasses. A single dispersal event within Acridocarpus is inferred from western Africa to eastern Africa between approximately 23 and 17 mya, coincident with the widespread replacement of humid forests by savannas in eastern Africa. Although the spread of xeric environments resulted in the extinction of many African plant groups, our data suggest that for others it provided an opportunity for further diversification. PMID- 12583581 TI - The neighborhood matters: effects of neighbor number and sibling (or kin) competition on floral traits in Spergularia marina (Caryophyllaceae). AB - Effects of abiotic factors on the expression of floral and gender traits have been well documented in wild plant species; by contrast, little is known of the effect(s) on an individual's floral phenotype of the genetic composition of its neighboring conspecifics. Here we report the results of a greenhouse experiment conducted to detect the effects of genetic attributes of an individual's local environment on the expression of floral traits in the selfing annual, Spergularia marina (Caryophyllaceae). First, to test the hypothesis that negative effects of intraspecific competition are stronger when an individual competes with genetically similar individuals than when it competes with unrelated genotypes, we evaluated the effects of the number of nearest-neighbor kin (vs. unrelated individuals) on floral traits. Plants adjacent to two kin produced significantly fewer stamens per flower than plants adjacent to one or no siblings, indicating that kin competition reduced allocation to male function. Second, to test the hypothesis that the genetic diversity of a neighborhood influences the phenotype of a focal plant, we determined whether the number of maternal families represented among an individual's nearest neighbors influences its phenotype. The number of maternal families surrounding a focal plant did not affect floral trait expression. These results suggest that in S. marina, male function is more sensitive than female function to the genetic environment and that as an individual's genetic similarity to its neighbors increases, so do the negative effects of competition for limited resources. PMID- 12583582 TI - Consequences of floral complexity for bumblebee-mediated geitonogamous self pollination in Salvia nipponica Miq. (Labiatae). AB - I address how floral complexity influences geitonogamous self-pollination through manipulation of pollinator behavior in Salvia nipponica. The pivoting stamens of S. nipponica hinder nectar-collecting bumblebees from crawling into flowers, increasing the probing time per flower. I predicted that longer probing times would reduce the relative cost of moving between plants, causing bees to leave plants earlier. To test this prediction, I simplified S. nipponica flowers by removing the stamens from all open flowers within a 75-m2 quadrat. Bumblebees probed these flowers more quickly than intact flowers, but the stamen removal affected neither the frequency of flower revisitation nor the flight distance between plants. In response to the decrease in the probing time per flower, bees probed more flowers on these plants. Therefore, in S. nipponica, floral complexity reduces the opportunity for geitonogamous self-pollination. Stamen removal also increased bee visitation per flower, suggesting that this sort of complexity deters visitation. To keep complex flowers attractive, therefore, selection might increase floral rewards or longevity. Floral complexity might evolve in an integrative manner with the rest of the floral phenotype. PMID- 12583583 TI - Sexual isolation between two sibling species with overlapping ranges: Drosophila santomea and Drosophila yakuba. AB - Drosophila yakuba is widespread in Africa, whereas D. santomea, its newly discovered sister species, is endemic to the volcanic island of Sao Tome in the Gulf of Guinea. Drosophila santomea probably formed after colonization of the island by a D. yakuba-like ancestor. The species presently have overlapping ranges on the mountain Pico do Sao Tome, with some hybridization occurring in this region. Sexual isolation between the species is uniformly high regardless of the source of the populations, and, as in many pairs of Drosophila species, is asymmetrical, so that hybridizations occur much more readily in one direction than the other. Despite the fact that these species meet many of the conditions required for the evolution of reinforcement (the elevation of sexual isolation by natural selection to avoid maladaptive interspecific hybridization), there is no evidence that sexual isolation between the species is highest in the zone of overlap. Sexual isolation is due to evolutionary changes in both female preference for heterospecific males and in the vigor with which males court heterospecific females. Heterospecific matings are also slower to take place than are homospecific matings, constituting another possible form of reproductive isolation. Genetic studies show that, when tested with females of either species, male hybrids having a D. santomea X chromosome mate much less frequently with females of either species than do males having a D. yakuba X chromosome, suggesting that the interaction between the D. santomea X chromosome and the D. yakuba genome causes behavioral sterility. Hybrid F1 females mate readily with males of either species, so that sexual isolation in this sex is completely recessive, a phenomenon seen in other Drosophila species. There has also been significant evolutionary change in the duration of copulation between these species; this difference involves genetic changes in both sexes, with at least two genes responsible in males and at least one in females. PMID- 12583584 TI - Insecticide resistance enhances male reproductive success in a beetle. AB - Malathion-specific resistance in the red flour beetle, Tribolium castaneum, is widespread and stable in natural populations even in the absence of pesticide exposure. To understand this stability, both resistant and susceptible males were placed in competition for susceptible female fertilization. Females were then isolated and their progeny was tested for malathion susceptibility. Male reproductive success was estimated for populations originating from different geographic areas and for isogenic strains. In most cases, resistant males had a greater reproductive success rate than susceptibles. The results suggest that male reproductive success is not traded against the selection for malathion resistance, even resistant individuals are at an advantage for this fitness trait. This absence of fitness cost may be the result of postselection of (1) modifier gene which ameliorate the fitness of resistant individuals or (2) nondeleterious resistance gene. Resistant phenotype superiority could be due to increased male mating success, improved ability of resistant males in sperm competition, female mate choice, and/or cryptic female choice of resistance gene(s). The effect of male phenotypic frequency on male reproductive success was also examined. We observed that male fertilization success is frequency dependent and inversely related to their frequency. However, this "rare male" advantage did not counteract the superiority of the resistant males. PMID- 12583585 TI - Speciation by host switch and adaptive radiation in a fish parasite genus Gyrodactylus (Monogenea, Gyrodactylidae). AB - Four hundred Gyrodactylus species have been formally described, but the estimated number of species in this fish ectoparasite genus of Monogenean Platyhelminthes is more than 20,000. The unusually high species richness has lead to the hypotheses of speciation and adaptive radiation via host switching. These hypotheses were tested by reconstructing a molecular phylogeny for the subgenus G. (Limnonephrotus) which is a group of freshwater parasites, including five species infecting wild and farmed salmonids. The highly variable ITS1 and ITS2 segments and the conservative 5.8S ribosomal gene were sequenced in 22 species plus two species representing the subgenus G. (Paranephrotus) as an outgroup. The phylogeny was compared with host systematics: the species were collected from six fish families (Cyprinidae, Salmonidae, Percidae, Esocidae, Gasterosteidae, and Gobitidae). The phylogenetic analysis demonstrated that G. (Limnonephrotus) is a monophyletic group that was originally hosted by cyprinids. The speciation has occurred in two episodes, the older one manifested in genetic distances 25-33% (4 6 Myr BP). The latter speciation burst occurred in one clade only, perhaps one million years ago. This clade has been morphologically identified as a wageneri species group. It is a monophyletic group of 18 species [studied here] and contains all five salmonid parasites, but also parasites, on cyprinids, percids, esocids, and gasterosteids. In G. (Limnonephrotus), eight host switches crossing the host family barrier were observed, and at least three of them were followed by repetitive speciation. Seven host-switch events were statistically confirmed by bootstrapping. The suggested model of speciation by host switch was accepted, and interestingly the adaptive radiation seems to be a consequence of host switch to a new family (key innovation model). The molecular and ecological evolution rate of Gyrodactylus parasites is manyfold in comparison to host species, and the phylogenies are largely independent and disconnected. PMID- 12583586 TI - Coevolution between Lamellodiscus (Monogenea: Diplectanidae) and Sparidae (Teleostei): the study of a complex host-parasite system. AB - Host-parasite coevolution was studied between Sparidae (Teleostei) fishes and their parasites of the genus Lamellodiscus (Monogenea, Diplectanidae) in the northwestern Mediterranean Sea. Molecular phylogenies were reconstructed for both groups. The phylogenctic tree of the Sparidae was obtained from previously published 16S mitochondrial DNA (mtDNA) sequences associated with new cytochrome b mtDNA sequences via a "total evidence" procedure. The phylogeny of Lamellodiscus species was reconstructed from 18S rDNA sequences that we obtained. Host-parasite coevolution was studied through different methods: TreeFitter, TreeMap, and a new method, ParaFit. If the cost of a host switch is not assumed to be high for parasites, all methods agree on the absence of widespread cospeciation processes in this host-parasite system. Host-parasite associations were interpreted to be due more to ecological factors than to coevolutionary processes. Host specificity appeared not to be related to host-parasite cospeciation. PMID- 12583587 TI - Temporal variation in divergent selection on spine number in threespine stickleback. AB - Short-term temporal cycles in ecological pressures, such as shifts in predation regime, are widespread in nature yet estimates of temporal variation in the direction and intensity of natural selection are few. Previous work on threespine stickleback (Gasterosteus aculeatus) has revealed that dorsal and pelvic spines are a defense against gape-limited predators but may be detrimental against grappling insect predators. In this study, we examined a 15-year database from an endemic population of threespine stickleback to look for evidence of temporal shifts in exposure to these divergent predation regimes and correlated shifts in selection on spine number. For juveniles, we detected selection for increased spine number during winter when gape-limited avian piscivores were most common but selection for decreased spine number during summer when odonate predation was more common. For subadults and adults, which are taken primarily by avian piscivores, we predicted selection should generally be for increased spine number in all seasons. Among 59 comparisons, four selection differentials were significant (Bonferroni corrected) and in the predicted direction. However, there was also substantial variability in remaining differentials, including two examples with strong selection for spine reduction. These reversals were associated with increased tendency of the fish to shift to a benthic niche, as determined from examination of stomach contents. These dietary data suggest that increased encounter rates with odonate predation select for spine reduction. Strong selection on spine number was followed by changes in mean spine number during subsequent years and a standard quantitative genetic formula revealed that spine number has a heritable component. Our results provide evidence of rapid morphological responses to selection from predators and suggest that temporal variation in selection may help maintain variation within populations. Furthermore, our findings indicate that variable selection can be predicted if the agents of selection are known. PMID- 12583588 TI - The evolution of development: two portraits of skull ossification in pipoid frogs. AB - Development creates morphology, and the study of developmental processes has repeatedly shed light on patterns of morphological evolution. However, development itself evolves as well, often concomitantly with changes in life history or in morphology. In this paper, two approaches are used to examine the evolution of skull development in pipoid frogs. Pipoids have highly unusual morphologies and life histories compared to other frogs, and their development also proves to be remarkable. First, a phylogenetic examination of skull bone ossification sequences reveals that jaw ossification occurs significantly earlier in pipoids than in other frogs; this represents a reversal to the primitive vertebrate condition. Early jaw ossification in pipoids is hypothesized to result from the absence of certain larval specializations possessed by other frogs, combined with unusual larval feeding behaviors. Second, thin-plate spline morphometric studies of ontogenetic shape change reveal important differences between pipoid skull development and that of other frogs. In the course of frog evolution, there has been a shift away from salamander-like patterns of ontogenetic shape change. The pipoids represent the culmination of this trend, and their morphologies are highly derived in numerous respects. This study represents the first detailed examination of the evolution of skull development in a diverse vertebrate clade within a phylogenetic framework. It is also the first study to examine ossification sequences across vertebrates, and the first to use thin-plate spline morphometrics to quantitatively describe ontogenetic trajectories. PMID- 12583589 TI - A statistical test of unbiased evolution of body size in birds. AB - Of the approximately 9500 bird species, the vast majority is small-bodied. That is a general feature of evolutionary lineages, also observed for instance in mammals and plants. The avian interspecific body size distribution is right skewed even on a logarithmic scale. That has previously been interpreted as evidence that body size evolution has been biased. However, a procedure to test for unbiased evolution from the shape of body size distributions was lacking. In the present paper unbiased body size evolution is defined precisely, and a statistical test is developed based on Monte Carlo simulation of unbiased evolution. Application of the test to birds suggests that it is highly unlikely that avian body size evolution has been unbiased as defined. Several possible explanations for this result are discussed. A plausible explanation is that the general model of unbiased evolution assumes that population size and generation time do not affect the evolutionary variability of body size; that is, that micro and macroevolution are decoupled, which theory suggests is not likely to be the case. PMID- 12583590 TI - Environmental influences on the evolution of growth and developmental rates in passerines. AB - The reasons why growth and developmental rates vary widely among species have remained unclear. Previous examinations of possible environmental influences on growth rates of birds yielded few correlations, leading to suggestions that young may be growing at maximum rates allowed within physiological constraints. However, estimations of growth rates can be confounded by variation in relative developmental stage at fledging. Here, we reestimate growth rates to control for developmental stage. We used these data to examine the potential covariation of growth and development with environmental variation across a sample of 115 North American passerines. Contrary to previous results, we found that growth rates of altricial nestlings were strongly positively correlated to daily nest predation rates, even after controlling for adult body mass and phylogeny. In addition, nestlings of species under stronger predation pressure remained in the nest for a shorter period, and they left the nest at lower body mass relative to adult body mass. Thus, nestlings both grew faster and left the nest at an earlier developmental stage in species with higher risk of predation. Growth patterns were also related to food, clutch size, and latitude. These results support a view that growth and developmental rates of altricial nestlings are strongly influenced by the environmental conditions experienced by species, and they generally lend support to an adaptive view of interspecific variation in growth and developmental rates. PMID- 12583591 TI - Female-biased sexual size dimorphism in the yellow-pine chipmunk (Tamias amoenus): sex-specific patterns of annual reproductive success and survival. AB - Sexual size dimorphism is ultimately the result of independent, sex-specific selection on body size. In mammals, male-biased sexual size dimorphism is the predominant pattern, and it is usually attributed to the polygynous mating system prevalent in most mammals. This sole explanation is unsatisfying because selection acts on both sexes simultaneously, therefore any explanation of sexual size dimorphism should explain why one sex is relatively large and the other is small. Using mark-recapture techniques and DNA microsatellite loci to assign parentage, we examined sex-specific patterns of annual reproductive success and survival in the yellow-pine chipmunk (Tamias amoenus), a small mammal with female biased sexual size dimorphism, to test the hypothesis that the dimorphism was related to sex differences in the relationship between body size and fitness. Chipmunks were monitored and body size components measured over three years in the Kananaskis Valley, Alberta, Canada. Male reproductive success was independent of body size perhaps due to trade-offs in body size associated with behavioral components of male mating success: dominance and running speed. Male survival was consistent with stabilizing selection for overall body size and body size components. The relationship between reproductive success and female body size fluctuated. In two of three years the relationship was positive, whereas in one year the relationship was negative. This may have been the result of differences in environmental conditions among years. Large females require more energy to maintain their soma than small females and may be unable to maintain lactation in the face of challenging environmental conditions. Female survival was positively related to body size, with little evidence for stabilizing selection. Sex differences in the relationship between body size and fitness (reproductive success and survival) were the result of different processes, but were ultimately consistent with female-biased sexual size dimorphism evident in this species. PMID- 12583592 TI - The evolution of alternative cryptic female choice strategies in age-structured populations. AB - Cryptic female choice is a potentially important aspect of the sexual selection process. According to the theory of sexual dialectics, postcopulation manipulation of relative male fertilization success can provide an avenue by which females can circumvent attempts by males to control female reproduction. Here I use stochastic models to investigate the evolution of cryptic female choice in populations with and without age structure. In populations without age structure, cryptic female choice will evolve only when (1) precopulatory mate choice by females is inefficient, (2) variation in male fitness is correlated with a trait upon which a female can base her choice of mates, and (3) the cost of multiple mating is not too high. In populations with age structure, similar conditions apply. However, selection sometimes favors females that employ alternative strategies of female choice at different ages. These results help to define the types of biological systems in which we should expect to see the evolution of cryptic female choice. They also illustrate that the evolution of choice strategies in females may be complex and may mirror in some important respects the evolution of alternative mating tactics in males. PMID- 12583593 TI - Mutual interest between the sexes and reproductive success in Drosophila pseudoobscura. AB - The pre-mating behavior of female Drosophila pseudoobscura has been considered passive and "coy" relative to more active, "ardent," and indiscriminate male behavior. To test whether this long-held view-the "received wisdom" about mating behavior in Drosophila-is really true we carried out observations on how often D. pseudoobscura females approached males prior to courtship and copulation. By including only virgin females and males in the experiments, we eliminated the possibility that males are "coy" due to sperm limitation and females flexibly "coy" due to male manipulations that may affect the duration of remating inhibition. We observed the movements of females and males in vials during the first five minutes of exposure to one another. Video records revealed females went toward males as frequently as males toward females; we inferred that females were as interested in males as males in females. The total number of offspring emerging as adults correlated significantly with mutual, precourtship interest of both males and females in their vial-mates and latency to copulation. Thus, we hypothesize that females in nature approach males, perhaps actively soliciting male courtship simply by remaining close to them. PMID- 12583594 TI - Direct and correlated responses to artificial selection on developmental time and wing length in Drosophila buzzatii. AB - Developmental time and body size are two positively correlated traits closely related to fitness in many organisms including Drosophila. Previous work suggested that these two traits are involved in a trade-off that may result from a negative genetic correlation between their effects on pre-adult and adult fitness. Here, we examine the evolution of developmental time and body size (indexed by wing length) under artificial selection applied to one or both traits in replicated D. buzzatii populations. Directional changes in both developmental time and wing length indicate the presence of substantial additive genetic variance for both traits. The strongest response to selection for fast development was found in lines selected simultaneously to reduce both developmental time and wing length, probably as an expected consequence of a synergistic effect of indirect selection. When selection was applied in the direction opposite to the putative genetic correlation, that is, large wing length but fast development, no responses were observed for developmental time. Lines selected to reduce both wing length and developmental time diverged slightly faster from the control than lines selected to increase wing length and reduce developmental time. However, wing length did not diverge from the control in lines selected only for fast development. These results suggest a complex genetic basis of the correlation between developmental time and wing length, but are generally consistent with the hypothesis that both traits are related in a trade-off. However, we found that this trade-off may disappear under uncrowded conditions, with fast-developing lines exhibiting a higher pre-adult viability than other lines when tested at high larval density. PMID- 12583595 TI - Sex and death: CHD1Z associated with high mortality in moorhens. AB - Sex ratios in clutches of moorhens (Gallinula chloropus) in Britain were measured on 83 chicks using the sex-linked CHD1 gene (Chromo-helicase/ATPase-DNA binding protein 1). Among birds, the female is the heterogametic sex (Z and W chromosomes), and the male is homogametic (two copies of the Z chromosome). We report variation among the PCR-amplified fragments of the CHD1Z, and the death of nearly all heterozygous male chicks (92%). In contrast, survivorship among females and homozygote males was 54-60%. Mortality in male heterozygotes was significantly higher than that of male homozygotes (P < 0.001). Chick and egg biometrics were not significantly different between these males. The CHD1Z was unlikely to be directly responsible but may have been hitchhiked by the causal gene(s). The observations appear to follow a classic underdominance (heterozygote inferiority) pattern, but raise the paradoxical question of why one form of the Z chromosome has not been fixed, as is expected from evolutionary theory. We discuss possible explanations and include a survey of British populations based on skin specimens. PMID- 12583596 TI - What cell lineages tell us about the evolution of spiralia remains to be seen. AB - Cell-lineage trees may contain information about spiralian phylogeny, as proposed by Guralnick and Lindberg (2001). Here we discuss this possibility further and conclude that the cell-division pattern must be known in greater detail and the coding methods refined before a possible phylogenetic signal can be identified. PMID- 12583597 TI - Comprehension of maternal-fetal exchange: a route to better health. PMID- 12583598 TI - Hormones and human trophoblast differentiation: a review. AB - In the human, fetal cytotrophoblastic cells play a key role in the implantation process and in placental development. With the progression of placentation, two pathways of differentiation lead to the formation of two distinct phenotypes. In the villous trophoblast (fusion phenotype), the trophblast differentiates from the fusion of mononuclear cytotrophoblastic cells into a syncytium, the syncytiotrophoblast. Bathing the maternal blood, the syncytiotrophoblast is involved in maternal-fetal exchanges and in placental endocrine functions. In the extravillous trophoblast (proliferative/invasive phenotype), the cytotrophoblastic cells proliferate and migrate into the decidua, remodeling the pregnant endometrium and its vasculature. This review summarizes our current knowledge of the key step of villous differentiation-the cell-cell fusion of the cytotrophoblastic cells--and on the invasion process of extravillous trophoblast. Experimental evidence demonstrates that the genetic differentiation/invasion programs of cytotrophoblastic cells could be modulated by their environment: oxygen, extracellular matrix, and soluble factors (cytokines, growth factors, and hormones). Cytotrophoblastic cells fusion and the functional differentiation of villous trophoblast are specifically stimulated by estradiol, glucocorticoids, and human chorionic gonadotropin (hCG) whereas progesterone is ineffective. Because these hormones are temporally secreted in large amounts and present at the fetomaternal interface, they are in good position to play a physiologic role in trophoblast differentiation. hCG may be important very early in pregnancy, when production of this glycoprotein is maximal, whereas estrogen increasingly produced by the fetoplacental unit and cortisol secreted from the fetal adrenal may be implicated in the end-stage maturation and aging of the trophoblast. PMID- 12583599 TI - Placental glucose transfer and fetal growth. AB - One of the primary regulators of maternofetal glucose transfer is the density of glucose transporter proteins in the placenta. These transporters, members of the GLUT gene family of facilitated-diffusion transporters, are embedded in the microvillous (maternal-facing) and basal (fetal-facing) membranes of the syncytiotrophoblast, the main placental barrier layer. Eight members of this family have been described in human placental tissue, but only GLUT1 protein has been identified in the syncytium, where its distribution is asymmetric. The microvillous membrane contains markedly more transporter than the basal, and, as a result, the basal membrane acts as the rate-limiting step in transplacental glucose transport; thus, changes in the density of basal membrane GLUT1 will have a significant impact on transplacental glucoseflux. What little is known about syncytial GLUT1 expression is restricted to factors associated with fetoplacental growth and metabolism; GLUT is inversely regulated by glucose concentration and basal membrane GLUT1 is positively regulated by insulin-like growth factor I, placental growth hormone, and hypoxia. In vivo, basal membrane GLUT1 is upregulated over gestation, increased in diabetic pregnancy, and decreased in chronic hypoxia, while microvillous membrane GLUT1 is unaffected. The contrast between in vitro and in vivo regulation and the specific changes in GLUT1 distribution suggest more complex regulatory interactions than those yet described. PMID- 12583600 TI - Transport and metabolism of amino acids in placenta. AB - In all mammalian species, the 20 amino acids of the genetic code are required for net protein accretion. The nutritional supply of amino acids for growth is defined as the net umbilical uptake of amino acids, representing the net transfer from maternal circulation, through the placenta and then to the fetus, of essential and non-essential amino acids. In considering the primary role of the placenta in the delivery of amino acids to the fetus for metabolism, it is important to consider the multiplicity of factors that may affect these overall delivery rates, including the activity and location of amino acid transporter systems, changes in these systems as gestation advances, effects of changes in placental surface area, uteroplacental blood flows, and maternal concentrations of amino acids. In this review, we discuss placental amino acid transport, the systems and their associated proteins, umbilical uptake data in animal and human studies, and amino acid transport in fetal growth restriction. Additionally, we discuss the current pool of thought concerning the mechanisms of placental amino acid transport as generated through in vitro vesicle studies and how they relate to the in vivo fluxes of animal studies. Finally, we discuss fetoplacental amino acid metabolism and specifically interorgan exchange. PMID- 12583603 TI - Possible role of placental leptin in pregnancy: a review. AB - Leptin was initially identified as an adipocyte-derived hormone that decreases food intake and body weight via its receptor in the hypothalamus. Subsequent animal studies revealed various physiologic functions of leptin. Leptin plays an essential role in reproduction by regulating gonadotropin-releasing hormone secretion from the hypothalamus. It also modulates glucose metabolism by increasing insulin sensitivity and activates the sympathetic nervous system. In humans, leptin is also produced by placental trophoblasts and is secreted into both the maternal and fetal circulation. Leptin production in the placenta is increased in pregnancies complicated with several pathologic conditions. Leptin gene expression in the placenta is augmented in severe preeclampsia, and maternal plasma leptin levels in severe preeclampsia are significantly higher than those in normotensive pregnant women. Leptin production in the placenta is also increased in diabetic pregnancy with insulin treatment. Furthermore, leptin is proposed to play a functional role in implantation by virtue of its stimulatory effect on matrix metalloproteinase expression in cytotrophoblast. Dysregulation of leptin metabolism and/or function in the placenta may be implicated in the pathogenesis of various disorders during pregnancy, such as recurrent miscarriage, gestational diabetes, intrauterine growth retardation, and preeclampsia. In this review, possible roles of placental leptin are discussed. PMID- 12583602 TI - Calcium-binding proteins: distribution and implication in mammalian placenta. AB - During gestation, transport by the placenta is solely responsible for nutrient supply to the developing fetus. In this context, calcium (Ca2+) transport machinery of the placenta thus represents the primary tissue site for regulating fetal Ca2+ homeostasis. In humans, the transplacental movements of Ca2+ increase dramatically during the last trimester of gestation, when fetal skeletal mineralization is at its highest. However, little is known about the exact mechanism of transport. Evidence suggests that some developmentally expressed cytosolic Ca(2+)-binding proteins (CaBPs) have an important role in regulating or shuttling cytosolic Ca2+ since they are endowed with a high affinity for Ca2+ (approximately 10(6) M(-1)). CaBPs belong to a large family of eukaryotic proteins containing a specific helix-loop-helix structure, referred to as the EF hand motif, which counts more than 200 members. Several of these CaBPs were identified in the placenta: CaBP9k, CaBP28k, CaBP57k, oncomodulin, S-100P, S 100alpha, and S-100beta. This review discusses the current views in this field to guide future investigations into the localization and functions of CaBPs during Ca2+ intracellular homeostasis in the placenta. PMID- 12583604 TI - Placental growth hormones. AB - Survival and development of the mammalian conceptus depends on a variety of factors. Fetal growth is controlled by genetic and environmental determinants that may limit the mother's capacity to provide an appropriate environment (e.g., space, nutrients, temperature). Exchanges between the mother and fetus take place within the placenta. Interestingly, despite the diversity of mammalian species in terms of placental structure and hormonal functions, placental size at term always correlates with birth weight, reflecting the essential role of this temporary organ. The placenta is the site of major endocrine activity, including synthesis of a broad range of steroid and peptide hormones, growth factors, cytokines, and other bioactive factors. Some of these are produced exclusively by the placenta, including chorionic gonadotropin, and growth hormone (GH)/prolactin like hormones. This article focuses on the expression, regulation, and physiologic role of placental GHs in mammalian species. Published data suggest that placental GHs are essential for adapting the maternal metabolism to pregnancy, for normal placental development, and therefore for fetal growth. PMID- 12583605 TI - Maternal-placental-fetal interactions in the endocrine regulation of fetal growth: role of somatotrophic axes. AB - Inadequate growth in utero is associated not only with adverse fetal, perinatal, and neonatal outcomes, but also with an altered propensity for disease later in life. Conversely, fetal overgrowth is also associated with increased medical risks for both mother and fetus. The interaction between the fetal genome and the intrauterine environment determines in great part how fetal growth will progress. The placental, maternal, and fetal somatotrophic axes (growth hormone/insulin like growth factor-1) play key roles in modulating this interaction. Experimental undernutrition in animal models has numerous effects over these axes and provides insight into understanding fetal growth and its abnormalities. This review addresses the contributions made by the placental, maternal, and fetal somatotrophic axes to the regulation of fetal growth. PMID- 12583601 TI - Lipid metabolism in pregnancy and its consequences in the fetus and newborn. AB - During early pregnancy there is an increase in body fat accumulation, associated with both hyperphagia and increased lipogenesis. During late pregnancy there is an accelerated breakdown of fat depots, which plays a key role in fetal development. Besides using placental transferred fatty acids, the fetus benefits from two other products: glycerol and ketone bodies. Although glycerol crosses the placenta in small proportions, it is a preferential substrate for maternal gluconeogenesis, and maternal glucose is quantitatively the main substrate crossing the placenta. Enhanced ketogenesis under fasting conditions and the easy transfer of ketones to the fetus allow maternal ketone bodies to reach the fetus, where they can be used as fuels for oxidative metabolism as well as lipogenic substrates. Although maternal cholesterol is an important source of cholesterol for the fetus during early gestation, its importance becomes minimal during late pregnancy, owing to the high capacity of fetal tissues to synthesize cholesterol. Maternal hypertriglyceridemia is a characteristic feature during pregnancy and corresponds to an accumulation of triglycerides not only in very low-density lipoprotein but also in low- and high-density lipoprotein. Although triglycerides do not cross the placental barrier, the presence of lipoprotein receptors in the placenta, together with lipoprotein lipase, phospholipase A2, and intracellular lipase activities, allows the release to the fetus of polyunsaturated fatty acids transported as triglycerides in maternal plasma lipoproteins. Normal fetal development needs the availability of both essential fatty acids and long chain polyunsaturated fatty acids, and the nutritional status of the mother during gestation has been related to fetal growth. However, excessive intake of certain long chain fatty acids may cause both declines in arachidonic acid and enhanced lipid peroxidation, reducing antioxidant capacity. PMID- 12583607 TI - Role of placenta in preeclampsia. AB - Preeclampsia, which manifests itself as hypertension, proteinuria, and edema in pregnancy, requires the presence of trophoblast tissue but not a fetus. It is characterized by abnormal trophoblast invasion of the spiral arteries of the decidua and myometrium leading to a failure to establish an adequate uteroplacental blood flow and, therefore, is thought to give rise to relatively hypoxic trophoblast tissue. This, in turn, may promote an exaggerated state of oxidative stress in the placenta. This hypoxia/oxidative stress may then further attenuate trophoblast invasion but also alters placental villous angiogenesis leading to a poorly developed fetoplacental vasculature with abnormal reactivity. Oxidative stress per se may also affect vascular reactivity, blood flow, and oxygen and nutrient delivery to the fetus, which ultimately may be compromised. The synthetic and transport functions of the syncytiotrophoblast may also be altered, and there is an increased rate of trophoblast apoptosis. The linkage among abnormal trophoblast invasion, trophoblast dysfunction, and the maternal disease remains unidentified. The presumptive humoral factor that is released by the preeclamptic placenta to cause maternal disease remains elusive. Current therapies to prevent preeclampsia aim toward preventing the maternal syndrome, not preventing the primary pathophysiology. PMID- 12583606 TI - Placental stress factors and maternal-fetal adaptive response: the corticotropin releasing factor family. AB - The placenta and its accessory membranes amnion and chorion undertake the role of intermediary barriers and active messengers in the maternal-fetal dialog. They synthesize, metabolize, and serve as target to numerous hormones that regulate maternal and fetal physiology during pregnancy. Among these factors, corticotropin-releasing factor (CRF) has been one of the more investigated in the last decade. Increasing evidence indicates that in the event of acute or chronic metabolic, physical, or infectious stress, maternal or fetal physiologic and pathologic conditions may influence placental secretion of CRF. The current opinion is that the placenta actually takes part in a stress syndrome by releasing CRF, which may help to influence uterine perfusion, maternal metabolism, fluid balance, and possibly uterine contractility, thereby protecting the fetus from a hostile environment. PMID- 12583609 TI - Management of limy bile syndrome: no therapy, laparotomy or endoscopic treatment? PMID- 12583610 TI - Myocarditogenic epitopes and autoimmune myocarditis. AB - Experimental autoimmune myocarditis is provoked by immunization with cardiac myosin. This animal model finally develops into dilated cardiomyopathy through repetitive myosin injections. To identify the myocardiogenic epitope, therefore, it is imperative not only to understand the mechanism of induction, but also to produce specific therapies, such as a blocking therapy to suppress the autoimmune process. Thus, we attempted to identify the myocarditogenic epitope using recombinant peptides. Beta-cardiac myosin heavy chain (CMHC) was amplified from rat mRNA by a reverse transcription polymerase chain reaction method. The PCR primers were designed to narrow the epitopic amino acid portion from each N terminal to C-terminal site. These PCR products were cloned into an E. coli expression vector to produce fusion proteins consisting of a Histidine-tag and a myosin peptide. The segment of amplified CMHC including the epitopic amino acid sequence to provoke moderate myocarditis in vivo was reported previously. Each peptide solution was emulsified in an equal volume of complete Freund's adjuvant and given as an immunization to 7-week-old rats. On day 21 after immunization, the rats were sacrificed, and the fresh heart was observed pathologically. Through this immunization, we could restrict the myocardiogenic site. Lastly, this peptide was found to be located on residues from 1,124 to 1,153. Using ELISA, the antibodies against myocarditogenic peptides were easily identified. Whether or not the antibody productivity is linked to myocarditogenecity is discussed. PMID- 12583611 TI - Endoplasmic reticulum stress and diabetes mellitus. AB - Pancreatic beta-cells are strongly engaged in protein secretion and have highly developed endoplasmic reticulum (ER). Proper folding of polypeptide into a three dimensional structure is essential for cellular function and protein malfolding can threaten cell survival. Various conditions can perturb the protein folding in the ER, which is collectively called ER stress. In order to adapt ER stress conditions, the cells respond in three distinct ways such as transcriptional induction of ER chaperones, translational attenuation, and ER-associated degradation (ERAD). However, when ER functions are severely impaired, the cell is eliminated by apoptosis via transcriptional induction of CHOP/GADD153, the activation of cJUN NH2-terminal kinase, and/or the activation of caspase-12. Recent studies have revealed that beta-cell is one of the most susceptible cells for ER stress, and ER stress-mediated apoptosis in beta-cells can be a cause of diabetes. A comprehensive understanding of the impact of the ER stress pathway in beta-cells and how it relates to the development of diabetes may contribute to provide new targets for the prevention and treatment of this disease. PMID- 12583612 TI - Anti-TNFalpha therapy for rheumatoid arthritis: an update. AB - Many studies have confirmed that the long term use of biological agents targeting TNFalpha in therapy for rheumatoid arthritis give rise to sustained improvement in symptoms and signs of disease provided the anti-TNF agent is efficacious and of low immunogenicity. The current regimes for infliximab 3 or 10 mg/kg infusion in combination with weekly oral methotrexate, or of subcutaneous etanercept 25 mg twice per week fulfil these criteria. D2E7, a 'human' antibody produced by phage display, has also demonstrated efficacy in clinical trials. It has recently emerged that anti-TNF therapy protects joints from structural damage. One year data for infliximab and methotrexate combination therapy suggest that this regime reduces disability. In early RA, etanercept acts more rapidly than methotrexate to decrease symptoms and retard progression of erosions. In conclusion, for patients with established and early RA, anti-TNF therapies set a new standard for symptom control and joint protection. PMID- 12583613 TI - Prognostic and clinical significance of newly acquired complete right bundle branch block in Japan Airline pilots. AB - OBJECTIVE: The purpose of this study was to evaluate the prognostic and clinical significance of newly acquired complete right bundle branch block (CRBBB) in airline pilots. PATIENTS: This study included pilots with acquired CRBBB, identified from a group of over 2,700 Japan Airline pilots. When the pilots applied for employment, a past medical history, physical examination, electrocardiogram, and chest radiograph were obtained. The pilots with ECG abnormality including CRBBB were not included in the study because of hiring requirements. RESULTS: Thirty-six pilots with CRBBB were identified between 1983 and 2002. All pilots with CRBBB were evaluated for the presence of ischemic heart disease by treadmill exercise testing, echocardiogram and exercise thallium scintigraphy. Twelve individuals underwent coronary angiography. The mean age of pilots was 44.4 +/- 5.8 years. The mean observation period was 10.9 +/- 5.7 years. For each of the 36 study subjects, Holter electrocardiogram and echocardiogram were obtained every 6 months after the CRBBB was detected. Exercise stress testing was performed every year. Exercise thallium scintigraphy was performed every 2 years to detect ischemic heart disease. During the observation period, two pilots stopped flying temporarily because of frequent ventricular premature beats and one pilot stopped flying permanentaly because of atrial fibrillation. During the follow-up period, no cardiovascular events were observed in pilots with CRBBB who had no underlying ischemic heart disease. CONCLUSION: Acquired CRBBB does not confer a poor prognosis, particularly in young men working as a pilot if there is no evidence of ischemia on exercise stress testing, echocardiography and exercise thallium scintigraphy. PMID- 12583614 TI - Clinical analysis of patients requiring long-term mechanical ventilation of over three months: ventilator-associated pneumonia as a primary complication. AB - OBJECTIVE: To evaluate the clinical features, etiology, and prognosis of patients who required long-term mechanical ventilation (LMV) of over three months for respiratory failure following underlying disease, and observation of their clinical course until death. PATIENTS: Thirty-seven patients (27 males, 10 females) treated in the internal and medical intensive care unit at Kawasaki Medical School Kawasaki Hospital over the 16-year period from April 1985 to March 2001 were retrospectively studied. RESULTS: Many of these patients were elderly males with respiratory disease such as pulmonary emphysema or old pulmonary tuberculosis, which had developed into acute respiratory failure resulting in respiratory tract infection and initiation of mechanical ventilation. The survival rates of one year, three years and five years after the start of mechanical ventilation were 60%, 30%, and 16%, retrospectively, and the prognoses were poor. Respiratory tract infection was the most common and serious complication. Specifically, ventilator-associated pneumonia (VAP) was a complication in 21 patients and also the main-cause of death. VAP was observed 2.3 years after the initiation of mechanical ventilation with significant differences in the following risk factors being observed between VAP (+) and VAP (-) groups: chronic obstructive pulmonary disease, duration of mechanical ventilation, prior antibiotics, aspiration of gastric contents and use of histamine-type II receptor antagonist. The causative pathogens of VAP were Pseudomonas aeruginosa and Staphylococcus aureus, which were frequently isolated from tracheal aspirates. All patients with VAP caused by MRSA died shortly after contracting the infection. CONCLUSIONS: This study has demonstrated that appropriate treatment for respiratory tract infections such as VAP and the prevention of nasocomial infection due to MRSA is of paramount importance for patients requiring long-term mechanical ventilation of over three months. PMID- 12583615 TI - Epstein-Barr virus infections of the central nervous system. AB - OBJECTIVE: Epstein-Barr virus (EBV), a lymphotropic herpes virus causing infectious mononucleosis (IM), also causes various central nervous system (CNS) infections. In the present study, EBV CNS infections were investigated. PATIENTS AND METHODS: For adult inpatients in our hospital and related hospitals between 1984-2002, CNS syndromes with IM symptoms were examined, and serologic positives were assessed according to established criteria. Polymerase chain reaction (PCR) was performed for cerebrospinal fluid (CSF) from seven patients. RESULTS: Ten patients with EBV-related CNS infections were found; their mean age was 36 years (20-79 years). The neurologic forms were as follows: acute encephalitis (4 patients), acute cerebellar ataxia (1), acute disseminated encephalomyelitis (ADEM) (2), myelitis (1), and meningitis (2). The PCR from CSF was positive in two patients with meningitis, one patient with ADEM, and one patient with encephalitis-associated chronic EVB infection. One case of encephalitis and another of relapsing ADEM were attributed to chronic EBV infection. CONCLUSION: Our study identified a variety of EBV-related CNS infections. EBV CNS infections are divided into two groups: 1) CNS syndromes associated with primary EBV or reactivated infection, and 2) those associated with chronic EBV infection; it is notable that in the former, diverse CNS syndromes including ADEM can occur, whereas in the latter, chronic or recurrent CNS syndromes are produced. PMID- 12583616 TI - Single low-dose treatment with praziquantel for Diphyllobothrium nihonkaiense infections. AB - OBJECTIVE: To investigate whether praziquantel is effective against diphyllobothriasis nihonkaiense at doses less than the conventional dose. PATIENTS AND METHODS: Fourteen Japanese men with diphyllobopthriasis nihonkaiense were treated with a single administration of praziquantel at 5-10 mg/kg. RESULTS: Twelve patients expelled a complete integrated Diphyllobothrium nihonkaiense with its scolex. One patient expelled an integrated D. nihonkaiense without its scolex, and one patient expelled 2 complete integrated D. nihonkaiense with their scolex, and an integrated D. nihonkaiense without its scolex. The lack of eggs in stool samples was confirmed in these 2 patients after the end of therapy. All patients had not expelled proglottides after the end of therapy for more than 1 year. No side effects were identified in any of the patients. CONCLUSION: All patients were considered to be cured. Our results indicate that the oral administration of a single dose of praziquantel at 5-10 mg/kg is effective and safe against D. nihonkaiense infections. PMID- 12583608 TI - Pathophysiology and maternal biologic markers of preeclampsia. AB - Preeclampsia-increased blood pressure and proteinuria appearing after the twentieth week of pregnancy--is a major cause of materal and neonatal morbidity, leading to iatrogenic prematurity. Several lines of evidence suggest that the disorder is owing to diminished invasion of spiral arteries by trophoblastic cells, followed by reduced perfusion of the fetoplacental unit and oxidative stress. These alterations, in the presence of maternal predisposition, lead to endothelial dysfunction and occurrence of the clinical syndrome of preeclampsia (multisystemic lesions). Although the pathophysiology of preeclampsia is still unknown, progress has been made during the past 10 yr, and the early identification of at-risk women with the use of biochemical; ultrasonographic; and, more recently, genetic susceptibility markers has been the subject of intense research. In the present review, markers of maternal predisposition, placental implantation, oxidative stress, vasomotor regulation, and endothelial dysfunction are investigated as candidate markers in the early prediction of preeclampsia. Unfortunately, at the present time, no marker has been proven to have a clinically useful predictive performance in the general pregnant population, and, therefore, more research in that area is warranted. PMID- 12583617 TI - Limy bile syndrome complicated with primary hyperparathyroidism. AB - Limy bile is a rare condition characterized by excessive precipitation of calcium carbonate in the gallbladder. Cases of complicated hyperparathyroidism are extremely rare. There is only one reported case, where serum and urine calcium levels were high. On the other hand, the presence of limy bile in the common bile duct is also very rare, and has been reported in only 20 cases. We report a patient with obstructive jaundice in whom the initial abdominal radiography showed limy bile in the gallbladder and common bile duct, and laboratory values showed a high serum calcium level and highly sensitive PTH (HS-PTH) level. PMID- 12583619 TI - Ruptured atherosclerotic plaque distant from maximal stenosis in acute myocardial infarction. AB - We describe a patient with acute myocardial infarction who showed ruptured plaque distant from the maximally stenotic lesion. In a 54-year-old male patient with acute antero-lateral myocardial infarction, coronary angiography showed a resolution of occlusive lesion with residual stenotic lesion in the middle portion of the left anterior descending artery (LAD) following t-PA administration. One month later, coronary angiography again disclosed significant stenosis of the middle LAD. Intravascular ultrasound revealed ruptured plaque that was located proximal to the maximally stenotic site which is generally considered as the culprit lesion. In this case, transient vessel occlusion occurred at the maximally stenotic site probably associated with plaque rupture distant from this lesion. PMID- 12583618 TI - Drug-induced hepatitis with autoimmune features during minocycline therapy. AB - A 25-year-old woman with no history of liver disease developed liver dysfunction associated with severe jaundice and general malaise following a prolonged therapy with minocycline for acne vulgaris. Serum anti-nuclear antibody was detected and immunoglobulin G level was elevated. Symptoms resolved and liver function normalized following minocycline discontinuation and corticosteroid administration. Our diagnosis was drug-induced hepatitis with autoimmune features, as liver histology revealed acute hepatitis. Drug-induced hepatitis should be considered when liver dysfunction or systemic symptoms develops during long-term minocycline therapy. PMID- 12583620 TI - Superior vena cava syndrome due to fibrosing mediastinitis histologically identical to xanthogranulomatous pyelonephritis. AB - We present herein a case of superior vena cava (SVC) syndrome caused by localized fibrosing mediastinitis (FM), which had histological features similar to xanthogranulomatous pyelonephritis (XGP). A 63-year-old woman presented with facial swelling 5 months after undergoing right nephrectomy for XGP. Radiologic investigations of the chest confirmed the presence of SVC obstruction due to an intraluminal tumor. The histological features of the tumor were consistent with those of FM and were very similar to those of XGP. Although the pathogenesis of neither FM nor XGP is known, some pathogenic process of FM and XGP may be the same. PMID- 12583621 TI - Acute reversible myocardial depression associated with sepsis. AB - A 30-year-old man was admitted to our hospital for left lobar pneumonia with septic shock. Acute left-sided heart failure became evident as sepsis developed. Echocardiography revealed diffuse severe hypokinesis of the left ventricle (LV) and a pulmonary artery catheter showed Forrester subset II hemodynamics. Along with amelioration of sepsis and decrease of the serum concentrations of tumor necrosis factor-alpha and interleukin-6, LV hypokinesis improved. It is suggested that the patient's heart failure may have been due to functional depression of myocardial contractility resulting from a direct effect of the cytokines towards the cardiomyocytes, the so-called "septic myocardial depression". PMID- 12583622 TI - Adrenal pseudocyst mimicking a pheochromocytoma found after a traffic accident. AB - We report a case of adrenal pseudocyst difficult to differentiate from pheochromocytoma. A right adrenal mass was found in a 41-year-old man after a traffic accident. Plasma level of renin was low, plasma level of aldosterone, and urine levels of aldosterone and catecholamines were high. Dynamic tests for primary aldosteronism and pheochromocytoma were negative. No abnormal uptake was shown in 131I-adosterol or 131I-MIBG scintigram. Adrenal venous sampling presented high levels of catecholamines. Laparoscopic adrenalectomy was performed. Histological examination revealed a pseudocyst with a thick fibrocollagenous wall. We discuss the mechanism of high catecholamine levels in the right adrenal vein. PMID- 12583623 TI - Nephrotic syndrome due to primary AL amyloidosis, successfully treated with VAD and subsequent high-dose melphalan followed by autologous peripheral blood stem cell transplantation. AB - Primary AL amyloidosis involves vital organs from the early phase of illness, resulting in a poor prognosis. We report a patient with nephrotic syndrome due to this type of amyloidosis, who was successfully treated with two courses of VAD (vincristine, doxorubicin and dexamethasone) and subsequent high-dose melphalan (140 mg/m2) with autologous stem cell support. Following the serial chemotherapy his proteinuria improved, and M protein became undetectable in both serum and urine. To avoid the progression of primary AL amyloidosis, intensive chemotherapy should be actively used when the general status and vital organ functions are well preserved. PMID- 12583624 TI - Delayed onset acute renal failure associated with Amanita pseudoporphyria Hongo ingestion. AB - A 66-year-old man with diabetes developed acute renal failure after ingestion of Amanita pseudoporphyria Hongo. Laboratory data showed acute nonoliguric renal failure. A renal biopsy showed acute tubular necrosis with glomerular minor abnormalities. He received hemodialysis treatment for 3 weeks and his renal function normalized 2 months after admission. We discuss the differences in acute renal failure caused by possible toxins of Amanita pseudoporphyria Hongo from that caused by other poisonous mushrooms. PMID- 12583625 TI - Cyclophosphamide-induced late-onset lung disease. AB - We report the case of a 58-year-old woman who was considered to have died of cyclophosphamide (CPA)-induced late-onset lung disease. She underwent a right sided mastectomy due to breast cancer at the age of 50 followed by daily administration of 50 mg CPA and 20 mg tamoxifen for 2 years. A refractory cough and dyspnea began at the age of 56. Chest radiographs revealed diffuse infiltrates and pleural thickening. Her vital capacity was markedly reduced. Corticosteroid therapy was ineffective. The disease rapidly progressed with occasional episodes of pneumothoraces to her death. Postmortem examination revealed pulmonary fibrosis with marked elastosis. PMID- 12583626 TI - Acute eosinophilic pneumonia with fine nodular shadows. AB - A 19-year-old woman presented with acute onset of cough and dyspnea. She started smoking two weeks before the appearance of symptoms. On admission, arterial blood gas analysis on room air breathing revealed PaO2 55 Torr. Chest roentgenogram and high resolution computed tomograms showed localized fine nodular shadows at the right lower lung field. Bronchoalveolar lavage fluid revealed a high eosinophil count. Eosinophil infiltration was also observed in transbronchial lung biopsy specimens. The final diagnosis was acute eosinophilic pneumonia (AEP). Although few reports have demonstrated diffuse fine nodular shadows in AEP, localized fine nodular shadows on chest roentgenogram and CT may sometimes be the sign of AEP especially in the early phase of the clinical course. PMID- 12583627 TI - Sarcoidosis with bilateral epididymal and testicular lesions. AB - We report a case of genital sarcoidosis that presented characteristic features on MRI. A 25-year-old man sarcoidosis patient with ocular and lung lesions presented a painful mass in the left scrotum together with systemic symptoms of fever, appetite loss, headache, and stomach-ache during the tapering of steroids. The patient was hypercalcemic, and this was thought to be the cause of his systemic symptoms. MRI showed multiple nodules of bilateral testes and enlargement of bilateral epididymis; the patient was diagnosed with testicular and epididymal lesions of sarcoidosis. An increased steroid dosage improved his hypercalcemia and genital lesions. PMID- 12583628 TI - Nephrotic syndrome in a patient with intravascular lymphomatosis. AB - The association of malignancy with nephrotic syndrome and renal histopathologic abnormalities is well documented. We report a case of intravascular lymphomatosis (IVL) presenting as nephrotic syndrome which was diagnosed by renal biopsy. Histological examination of the renal biopsy specimens showed dissemination of neoplastic lymphoid cells throughout the glomerular capillary bed. These tumor cells were positive for CD20. Since the nephrotic syndrome improved with treatment of the lymphoma, intravascular lymphomatosis may well have played an important role in the pathogenesis of minimal change nephrotic syndrome in this patient. PMID- 12583629 TI - Autoimmune neutropenia associated with multiple sclerosis. AB - A 53-year-old Japanese man with multiple sclerosis developed autoimmune neutropenia. The neutrophil count was consistently less than 0.2 x 10(9)/l, irrespective of the disease activity of multiple sclerosis or the administration of immunosuppressive agents or granulocyte colony-stimulating factor. After high dose gamma-globulin therapy was started, temporary increases in the neutrophil count were observed. Despite a wide spectrum of clinical manifestations in multiple sclerosis, autoimmune neutropenia has never been reported previously. PMID- 12583630 TI - Asian variant of CD5+ intravascular large B-cell lymphoma with splenic infarction. AB - A 57-year-old man was admitted with fever and epigastralgia, and presented with splenomegaly and pancytopenia. A CT scan revealed splenic infarctions. There were no lymphadenopathies, skin lesions, or neurological abnormalities. A splenectomy was performed. Bone marrow involvement with hemophagocytosis was noted. The diagnosis of Asian variant of intravascular diffuse large B-cell lymphoma was based on intravascular and sinusoidal distribution of large CD5+ B cells. The patient died of the disease 11 months after onset. To our knowledge, this is the first report of AIVL that presented with splenic infarction. This distinct lymphoma should be included in the differential diagnosis of splenic infarction. PMID- 12583631 TI - Mesenteric venous thrombosis in hereditary protein C deficiency with the mutation at Arg169 (CGG-TGG). AB - A 39-year-old man with no significant medical history was admitted to our hospital with severe abdominal pain and melena. Computed tomographic (CT) scans demonstrated superior mesenteric venous thrombosis. Although thrombolysis and anticoagulant therapy was started immediately, symptoms of strangulation ileus developed. Laparotomy was therefore performed and revealed necrotic stenosis of the ileum. The patient, his father and sisters showed low protein C levels. Direct sequencing analysis of their protein C gene revealed a heterozygous mutation at codon 169 corresponding to the cleavage site of the activation peptide, which was referred to as protein C Tochigi. PMID- 12583632 TI - Intravascular large B cell lymphoma diagnosed by senile angioma biopsy. AB - An 82-year-old man without notable medical history was admitted to our hospital following subacute deterioration of apettite, disorientation and strange behavior. There was spasticity of the right extremities without weakness. LDH and serum soluble IL2 receptor antibody levels were elevated, and as well as the protein level and IgG levels in the cerebrospinal fluid. CT scanning of the brain revealed a lesion at the left corona radiata. The patient's level of consciousness was worsening, and follow-up study of the brain showed a new lesion in the left occipital lobe. Intravascular lymphomatosis was therefore suspected. We performed a skin biopsy from two typical senile angiomas. In one of these biopsy specimens, a capillary hemangioma was present in the mid-reticular dermis and it was filled with abnormal B cells. The diagnosis of intravascular B cell lymphoma (IVL) was thus established. IVL is a rare subtype of extranodal diffuse large B cell lymphoma with a poor outcome. However, it is recently thought that if the diagnosis is established early, aggressive chemotherapy increases survival. Senile angioma is a skin eruption that is considered prevalent for the most part in elderly people. If a patient is suspected to have IVL, and there is no appropriate site of biopsy, it might be beneficial to try a skin biopsy aiming at senile angiomas for early diagnosis. PMID- 12583633 TI - Amyloid arthropathy resembling seronegative rheumatoid arthritis in a patient with IgD-kappa multiple myeloma. AB - A 67-year-old woman suffered from symmetrical polyarthralgia and multiple joint swelling simulating rheumatoid arthritis (RA). Laboratory examination showed negative results for rheumatoid factor, decreased levels of IgG, IgA, and IgM, and an increased level of IgD. Immunoelectrophoresis in her serum and urine revealed an IgD-kappa monoclonal component and Bence Jones protein (kappa), respectively. A bone marrow biopsy showed an excess of atypical plasma cells. A synovial biopsy revealed amyloid deposition composed of IgD-kappa. She was diagnosed with amyloid arthropathy (AmyA) secondary to IgD-kappa multiple myeloma. It is important to pay attention to AmyA due to multiple myeloma in patients with seronegative RA. PMID- 12583634 TI - Idiopathic pancreatitis may be associated with ulcerative colitis. PMID- 12583635 TI - [123I] meta-iodobenzylguanidine myocardial scintigraphy differentiates corticobasal degeneration from Parkinson's disease. PMID- 12583636 TI - Disseminated mucormycosis in an acquired immunodeficiency syndrome (AIDS) patient. PMID- 12583637 TI - Cerebral toxoplasmosis in an AIDS patient. PMID- 12583638 TI - Editorial: quality in managed care? PMID- 12583640 TI - Commentary: presenting the value of medical quality to nonclinical senior management and boards of directors. AB - Many physicians find hospital or health plan boards of directors to be intimidating arenas for medical quality presentations. This essay presents a number of "pearls" gleaned from successful senior clinician managers who have learned to relate to senior management and advance in their careers. This commentary was developed from research and a presentation of the same title delivered at the American College of Medical Quality Annual Meeting held in Las Vegas in October, 2001. It is important that medical directors who work with financial managers convert quality concepts into "business value" concepts. Talking in the language of business, rather than the language of doctors, makes it much easier to communicate with management (although some translation is often in order). As a clinician presenting to financial managers, you should become familiar with financial terms and how they are used. Indeed, the development of a financial model representing clinical activity results in the highest level of success. There are a number of methods for estimating impact that have been found within general business and health services research areas that are acceptable. Successful presenters of information approach their task effectively. Reports are in a more readable format and convey information for action by the corporation rather than as a scholarly treatise. Approaching senior management, one must consider the psychology of individuals in senior positions. Senior medical executives who are successful report similar approaches to their tasks, and offer helpful insight into career advancement. PMID- 12583639 TI - Costs of measuring outcomes of acute hospital care in a longitudinal outcomes measurement system. AB - It is widely acknowledged that the measurement of outcomes of care and the comparison of outcomes over time within health care providers and risk-adjusted comparisons among providers are important parts of improving quality and cost effectiveness of care. However, few studies have assessed the costs of measuring outcomes of care. We sought to evaluate the personnel and financial resources spent for a prospective assessment of outcomes of acute hospital care by health professionals in internal medicine. The study included 15 primary care hospitals participating in a longitudinal outcomes measurement program and 2005 patients over an assessment period with an average duration of 6 months. Each hospital project manager participated in a previously-tested structured 30-minute telephone interview. Outcome measures include time spent by the individual job titles in implementing and running the outcomes measurement program. Job-title specific times were used to calculate costs from the hospitals' perspective. One time costs (2132 +/- 1352 Euros) and administrative costs (95 +/- 97 Euros per week) varied substantially. Costs per patient were fairly stable at around 20 Euros. We estimated that the total cost for each hospital to assess outcomes of care for accreditation (10 tracer diagnoses over 6 months) would be 9700 Euros and that continuous monitoring of outcomes (5 tracer diagnoses) would cost 12,400 Euros per year. This study suggests that outcomes of acute hospital care can be assessed with limited resources and that standardized training programs would reduce variability in overall costs. This study should help hospital decision makers to estimate the necessary funding for outcomes measurement initiatives. PMID- 12583642 TI - Federal and state nursing facility websites: just what the consumer needs? AB - Since the introduction of the Medicare Nursing Home Compare website in 1999, some states have begun to develop their own websites to help consumers compare nursing facilities (NFs). This article presents a brief conceptual framework for the type of information needed for an Internet-based information system and analyzes existing federal and state NF websites, using data collected from a survey completed in 2002. Twenty-four states and the District of Columbia have a variety of information on NFs, similar to the information on the Medicare website. Information on characteristics and deficiencies of a facility is the most commonly available, but a few states have data on ownership, staffing indicators, quality indicators, complaints, and enforcement actions. Other types of data, such as resident characteristics, staff turnover rates, and financial indicators, are generally not available. Although many states are making progress toward providing consumers with information, there are gaps that exist, which if filled, could provide consumers with a better tool for facility selection and monitoring the quality of care. PMID- 12583641 TI - Mental Health QUERI Initiative: expert ratings of criteria to assess performance for major depressive disorder and schizophrenia. AB - The purpose of this study was to examine mental health care experts' opinions about performance measures and associated data elements that could serve as the basis of an information system for monitoring the implementation of clinical practice guidelines for major depressive disorder (MDD) and schizophrenia in the Veterans Health Administration (VHA). Nineteen mental health care experts rated the meaningfulness and validity of performance measures and automated data elements. For MDD, experts rated the following measures as very meaningful and valid: (a) the results of depression screening, (b) the occurrence and results of diagnostic assessment for MDD, (c) the provision of antidepressant medication or psychotherapy, and (d) whether antidepressant medications were prescribed within the therapeutic dose range recommended by practice guidelines. For schizophrenia, expert reviewers rated therapeutic doses of antipsychotic medication and assessment for antipsychotic medication side effects as being very meaningful and valid. Performance measures that evaluate clinically significant aspects of care using specific, valid data elements are the most meaningful. Translation efforts by the VHA's Mental Health Quality Enhancement Research Initiative program include additional studies of the validity of some of the proposed data elements and the development of national clinical reminders for performance measures judged to be meaningful. PMID- 12583643 TI - Administrative data versus corrected administrative data. AB - The purpose of this research was to provide insight into the use of existing administrative data and to identify changes that could be made to improve broad based use of administrative data. Data were collected on patients hospitalized with pneumonia at a 715 bed hospital in North Carolina in 1996-1997. Patients were selected from administrative databases via diagnosis and charge codes. Outcome variables were length of stay and total hospital charges. Explanatory variables were age, sex, race, insurance type, season of year, admission source (emergency department or other), comorbidity score, care path designation, physician specialty and teaching appointment. These data were collected from administrative data and then from a limited chart review to correct the administrative data. We found no significant differences in economic outcomes between the administrative data and the corrected administrative data. Administrative data appear to be a reliable and cost-effective data source for quality assessment. PMID- 12583644 TI - Epidural analgesia and severe perineal laceration in a community-based obstetric practice. AB - BACKGROUND: This study assessed whether epidural analgesia was an independent risk factor for severe perineal laceration. METHODS: A retrospective cohort study analyzed 2,759 patients at St. Francis Regional Medical Center who had vertex, spontaneous or induced, singleton, live, vaginal deliveries of neonates of at least 36 weeks' gestation. Patients with diabetes or severe cardiac disease were excluded. Outcomes measured were third- or fourth-degree perineal lacerations. RESULTS: Overall rate of severe perineal laceration was 6.38% (n = 176). Epidural analgesia was given to 634 (22.98%) women. Among women who had epidural analgesia, 10.25% (65 of 634) had severe perineal lacerations compared with 5.22% (111 of 2,125) of the women who did not have epidural analgesia. After controlling for major variables in a logistic regression analysis, epidural analgesia remained a significant predictor of severe perineal injury (odds ratio [OR] = 1.528, 95% confidence interval [CI] = 1.092-2.137). When instrument use was included in the model, epidural analgesia was no longer a statistically significant, independent predictor of severe perineal injury. (OR = 1.287, 95% CI = 0.907-1.826). Instrument use was found to be a strong predictor of severe laceration (OR = 3.245, 95% CI = 2.162-4.869). A logistic regression model examining predictors of instrument use found that epidural analgesia does significantly predict instrument use (OR = 3.01, 95% CI = 2.225-4.075). CONCLUSION: Epidural analgesia is associated with an increase in severe perineal trauma as a result of an associated threefold increased risk of instrument use. Instrument use in vaginal delivery more than triples the risk of severe perineal laceration. PMID- 12583645 TI - Effect of labor epidural anesthesia on breast-feeding of healthy full-term newborns delivered vaginally. AB - BACKGROUND: Epidural anesthesia is commonly administered to laboring women. Some studies have suggested that epidural anesthesia might inhibit breast-feeding. This study explores the association between labor epidural anesthesia and early breast-feeding success. METHODS: Standardized records of mother-baby dyads representing 115 consecutive healthy, full-term, breast-feeding newborns delivered vaginally of mothers receiving epidural anesthesia were analyzed and compared with 116 newborns not exposed to maternal epidural anesthesia. Primary outcome was two successful breast-feeding encounters by 24 hours of age, as defined by a LATCH breast-feeding assessment score of 7 or more of 10 and a latch score of 2/2. Means were compared with the Kruskal-Wallis test. Categorical data were compared using the Mantel-Haenszel chi-square test. Stratified analysis of potentially confounding variables was performed using Mantel-Haenszel weighted odd ratios (OR) and chi-square for evaluation of interaction. RESULTS: Both epidural and nonepidural anesthesia groups were similar except maternal nulliparity was more common in the epidural anesthesia group. Two successful breast-feedings within 24 hours of age were achieved by 69.6% of mother-baby units that had had epidural anesthesia compared with 81.0% of mother-baby units that had not (odds ratio [OR] 0.53, P = .04). These relations remained after stratification (weighted odds ratios in parenthesis) based on maternal age (0.52), parity (0.58), narcotics use in labor (0.49), and first breast-feeding within 1 hour (0.49). Babies of mothers who had had epidural anesthesia were significantly more likely to receive a bottle supplement while hospitalized (OR 2.63; P < .001) despite mothers exposed to epidural anesthesia showing a trend toward being more likely to attempt breast-feeding in the 1 hour (OR 1.66; P = .06). Mothers who had epidural anesthesia and who did not breast-feed within 1 hour were at high risk for having their babies receive bottle supplementation (OR 6.27). CONCLUSIONS: Labor epidural anesthesia had a negative impact on breast feeding in the first 24 hours of life even though it did not inhibit the percentage of breast-feeding attempts in the first hour. Further studies are needed to elucidate the exact nature of this association. PMID- 12583646 TI - Underdiagnosis of obesity at a community health center. AB - BACKGROUND: Obesity is at epidemic proportions. This study examined the extent to which obesity is being diagnosed at a community health center residency-training site. Results were examined by provider type. Characteristics of patients with obesity diagnosed by primary care providers were compared with characteristics of patients determined to be obese by body mass index (BMI) calculation exclusively. METHODS: A cross-sectional design was used. Medical records of 465 adult patients were audited. Data collected included diagnosis of obesity, height and weight, demographics, and comorbidity. RESULTS: Of the 465 patients' charts audited, 83 contained a provider diagnosis of obesity, and 74 additional patients were determined to be obese by BMI calculation exclusively. Significant underdiagnosis occurred among all provider types (P = .036). Patients with a diagnosis of obesity had significantly higher BMI scores (38.4 vs 34.4, P = .002). Obesity was more likely to be diagnosed in female than in male patients (P = .001). Differences related to age, insurance coverage, and comorbidity were not significant. CONCLUSIONS: Obesity was found to be an underdiagnosed condition among all provider types. As evidenced by significantly higher BMI scores for provider-diagnosed obesity, the data suggest that the obesity diagnosis is made by appearance. The importance of teaching and modeling the use of BMI to diagnose obesity is underscored. PMID- 12583647 TI - Patient beliefs predict response to paroxetine among primary care patients with dysthymia and minor depression. AB - BACKGROUND: Dysthymia and minor depression are common problems in primary care, but it is not known how patient health beliefs shape response to antidepressant treatment of these less severe forms of depression. METHODS: Three hundred thirty three primary care patients with dysthymia or minor depression received at least 4 weeks of paroxetine or placebo in a multicenter, randomized controlled 11-week trial. Patient health beliefs and other characteristics were examined as predictors of treatment adherence and depression remission. RESULTS: Patient beliefs were not predictive of adherence to paroxetine or placebo. Patients with less endorsement of biological beliefs about their condition (odds ratio [OR] = 3.40), higher perceived general health (OR = 3.38), meeting criteria for dysthymia (OR = 2.37), and age younger than 60 years (OR = 2.68) were more likely to achieve remission on paroxetine. Patient beliefs did not predict remission on placebo. Those with lower severity of depression symptoms at baseline (OR = 2.70) and women (OR = 2.18) were most likely to achieve remission on placebo. CONCLUSIONS: Our results suggest that patients with dysthymia or minor depression are more likely to respond to antidepressant medication if they do not see their depression as a biological illness and see themselves as generally healthy. It is clearly not necessary for patients to believe that their dysthymia or minor depression is biological to respond to antidepressant medication. PMID- 12583648 TI - Violent victimization of women and men: physical and psychiatric symptoms. AB - BACKGROUND: Our objectives for this exploratory study were (1) to assess the prevalence in a family practice of violent victimization of women and men by partners, friends, families, and strangers, and (2) to compare the physical symptoms, depression, alcohol use problems, and social support of women and men who were or were not victimized in the previous 12 months. METHODS: We conducted a cross-sectional, multicenter study of family practice patients (1999-2000). One thousand twenty-four patients, including 679 women and 345 men from 18 to 64 years of age completed a standard health history and a demographic questionnaire. The health history questionnaire included a question about violent victimization. RESULTS: Violent victimization was reported by 9.9% of the women and 10.9% of the men. Patients who were victimized were grouped into those who were victimized by partners (4.9% of women and 3.0% of men); by friends, or family, or strangers (2.3% of women and 5.0% of men); or by more than one category of persons other than partners (2.6% of women and 3.0% of men). Almost one third of patients victimized by partners were also victimized by another person. Women who were victimized had more physical symptoms than women who were not victimized. Women who were victimized and men who were victimized by their partners had more depressive symptoms than other women and men. Patients who were victimized by more than one category of other victimizers reported more alcohol use problems than other patients. Patients who were victimized reported less social support than patients who were not victimized. CONCLUSIONS: Both women and men report violent victimization in response to a screening question. Violence by partners and by others is related to physical and psychiatric symptoms in women and in men. PMID- 12583649 TI - Improving firearm storage habits: impact of brief office counseling by family physicians. AB - BACKGROUND: Firearm injury is the leading cause of injury-related death among youth and second leading cause of injury-related death overall in the United States. Our objective is to determine the impact of brief office counseling by family physicians on patients' firearm storage habits. METHODS: Of the 1,233 patients who completed the enrollment questionnaire, 156 (13%) reported they had guns in their household and agreed to participate in the study. Postintervention survey instruments were completed by 127 (81%) of participants. Participants received either no counseling, verbal counseling alone, or counseling and a gun safety brochure from their physician. Firearm storage habits were measured at baseline and 60 to 90 days after intervention. RESULTS: At the postintervention interview, 64% of the group receiving verbal counseling and 58% of the group receiving verbal counseling plus written information made a safe change in gun storage compared with 33% of participants in the no-intervention group (P =. 02). A logistic regression model controlling for demographics and gun ownership showed that compared with the no-intervention group, intervention participants were three times more likely to make safe changes. CONCLUSIONS: Family physicians' brief counseling efforts made a significant positive impact in the firearm storage habits of their patients. With a verbal or written recommendation, a significant improvement was observed in firearm storage. PMID- 12583651 TI - End-of-life care for a man with developmental disabilities. AB - BACKGROUND: Science can artificially maintain many essential life functions. Does such care prolong life or dying? METHODS: A case is described of a patient with developmental disability with unknown health care choices who was hospitalized for drug-resistant urosepsis. He developed aspiration pneumonia, deep vein thrombosis, and respiratory arrest. He required gastrostomy, tracheostomy, artificial ventilation, parenteral nutrition, hemodialysis, multiple anti infective agents, and blood transfusions. On day 58, a bioethics committee recommended against cardiopulmonary resuscitation. On day 66, the patient's conservator concurred but required continuation of artificial ventilation. To the dismay of some caretakers, the patient continued to receive intrusive care until his death on day 104. The hospital charge was $709,206. RESULTS AND CONCLUSION: Hospital care of patients with mental incapacity can be clinically and ethically challenging. End-of-life decisions can be facilitated when the patient's legal representative and physician actively advocate the patient's best interests and communicate frequently and openly. Suggestions are made for such exigencies. PMID- 12583650 TI - Botanicals and dietary supplements in diabetic peripheral neuropathy. AB - BACKGROUND: Many persons use botanicals and dietary supplements for chronic conditions that do not respond to traditional Western medications. Tricyclic antidepressants, a common treatment option for diabetic neuropathy, can have many side effects and are a poor choice in certain populations (eg, the elderly). As such, patients might turn to botanicals and dietary supplements, not realizing that these products are not well regulated. METHODS: This article reviews botanicals and dietary supplements that have been involved in randomized controlled trials (RCTs) for diabetic neuropathy. We searched MEDLINE for English language literature dating from 1966 to April 2001 using the following subject headings: (1) diabetes and botanical, herb, and supplement, (2) neuropathy and botanical, herb, and supplement, and (3) diabetic neuropathy and botanical, herb, and supplement. RESULTS: Our search found agents that might improve symptoms of neuropathy (eg, evening primrose oil, alpha-lipoic acid, capsaicin) without affecting glucose control. Botanicals and dietary supplements involved in only one RCT or associated with little clinical benefit were reviewed in brief. CONCLUSIONS: Evening primrose oil, alpha-lipoic acid, and capsaicin have received the greatest attention for their use in diabetic neuropathy, but further studies are needed to confirm their efficacy. Patients using these products need to be informed of potential drug interactions and side effects. PMID- 12583653 TI - A Lisfranc fracture-dislocation in a football player. PMID- 12583654 TI - Postcardiac injury syndrome after coronary angioplasty and stenting. PMID- 12583652 TI - Hydrops fetalis secondary to parvovirus B19 infections. AB - BACKGROUND: Fetal infection by human parvovirus B19 is a common cause of fetal anemia, nonimmune hydrops fetalis, and spontaneous abortion and can result in fetal death. Recent improvements in diagnosing parvovirus infections and the availability of intrauterine transfusion have reduced the overall rate of fetal loss after maternal exposure. METHODS: We report two cases of maternal parvovirus infection with classic findings of hydrops fetalis and review various aspects of parvovirus infection with emphasis on the developing management options in pregnancy. RESULTS AND CONCLUSIONS: Different management led to different results. In the first case there was normal neonatal and infantile development, and in the second case, the fetus died. With accurate laboratory testing, obstetric sonography, and fetal transfusion, the fetal mortality from parvovirus infection has been reduced considerably, and most pregnancies complicated by maternal parvovirus infection result in healthy outcomes. PMID- 12583655 TI - Quest for success: Nicholas Pisacano Scholars presentation. PMID- 12583657 TI - Family interdependencies and the family physician. PMID- 12583656 TI - The challenge of caring for the severely disabled. PMID- 12583659 TI - Use of drug sample medications. PMID- 12583660 TI - Sample medications and the uninsured. PMID- 12583661 TI - Canadian health care system. PMID- 12583662 TI - Operation infinite injustice: impact of sanctions and prospective war on the people of Iraq. PMID- 12583663 TI - Alcohol and university student drinking--not a class act. PMID- 12583664 TI - Heavy drinking on Canadian campuses. AB - OBJECTIVE: To describe the prevalence and frequency of heavy drinking episodes among Canadian undergraduates. METHODS: Data are drawn from the Canadian Campus Survey, a national mail survey, conducted in the fall of 1998, with a random sample of 7,800 students from 16 universities. RESULTS: Overall, 62.7% and 34.8% of students reported consuming 5 or more drinks and 8 or more drinks, respectively, on a single occasion at least once during the fall semester. On average, drinkers reported having 5 or more drinks almost 5 times during the fall semester, and having 8 or more drinks almost twice during the same period. The groups reporting the highest rates of heavy drinking were males, those living in university residences, those with low academic orientation and those with high recreational orientation. INTERPRETATION: Generally, this study has shown that heavy drinking is highly engrained in Canadian undergraduates' drinking patterns, and is related to a number of factors. These factors can be used to develop targeted prevention efforts. PMID- 12583665 TI - Cigarette use among Canadian undergraduates. AB - OBJECTIVE: To describe the prevalence of daily cigarette use among Canadian undergraduates. Estimates are also compared to earlier Ontario surveys. METHODS: Data are drawn from the Canadian Campus Survey, a national mail survey, conducted in the fall of 1998, with a random sample of 7,800 students from 16 universities. RESULTS: Overall, 17.1% reported daily cigarette smoking and 10.4% reported occasional smoking. Rates of daily smoking differed significantly by region (with rates above average among those attending university in the Atlantic and below average among those attending university in British Columbia and the Prairies), residence (those residing off campus without family reported the highest prevalence rate), and year of study (those in the final year typically reported lower rate of use). INTERPRETATION: University campuses represent an environment with potential gains to be made by tobacco control policies. PMID- 12583666 TI - The Canadian slave smoking machine. PMID- 12583667 TI - Smoking, physical activity, and diet in North American youth: where are we at? PMID- 12583668 TI - The early 1990s cigarette price decrease and trends in youth smoking in Ontario. AB - BACKGROUND: Youth are especially vulnerable to fluctuations in cigarette price, and both the smuggling increase during the early 1990s and the 1994 tax decrease made purchasing easier for youth. The purpose of this study is to examine the relation between these price decreases, and trends in smoking prevalence and amount smoked among Ontario youth. METHODS: Data from the Ontario Student Drug Use Survey were analyzed for trend using: 1) polynomial regression, and 2) discontinuity regression with an "event time" of 1993 to capture effects of both pre-tax cut smuggling and the tax cut. RESULTS: Overall, smoking prevalence decreased from 1977 to 1993, jumped upward at this time, and decreased after 1993. Among daily smokers, mean number of cigarettes smoked daily showed an increase followed by a decrease over the 24 years, and a negative quadratic trend. Trends for subgroups are also reported. CONCLUSIONS: These findings suggest that the early 1990s cigarette price decrease may have played a role in increasing youth smoking in Ontario. PMID- 12583669 TI - Support for tobacco control policies: how congruent are the attitudes of legislators and the public? AB - OBJECTIVES: To examine the congruence in perceptions and attitudes of legislators and the public regarding tobacco and tobacco control policies. METHODS: Two cross sectional surveys were used, one of elected federal and provincial legislators and one of adult residents in Ontario, Canada. Perceptions and attitudes were analyzed as dependent variables using multiple logistic regression, and adjusted for age, sex, educational attainment, and smoking status. FINDINGS: Congruence was found in most instances, however, some differences were found. Legislators were more likely than the public to agree that most smokers are addicted and were more supportive of a smoking ban in workplaces, but these differences disappeared after controlling for socio-demographic characteristics. Legislators were also more aware than the public of the magnitude of deaths due to tobacco compared to alcohol, whereas the public was more supportive of strong penalties against stores that sell cigarettes to minors. CONCLUSIONS: Our findings provide considerable evidence for congruence in the "real-world" (unadjusted) perceptions and attitudes of Ontario legislators and the Ontario public toward tobacco control policies. Such findings are positive for tobacco control advocates and should be leveraged to bring forward strong tobacco policies in the political arena. PMID- 12583670 TI - Immunization initiatives. Polio...the finish line is in sight! PMID- 12583678 TI - Smoking, physical activity and breakfast consumption among secondary school students in a southwestern Ontario community. AB - OBJECTIVES: To determine the prevalence of smoking, low levels of physical activity, and missing breakfast among students (n=318) in grades 9 through 12 in three schools in southwestern Ontario; to see if these behaviours were associated; and, whether there were gender differences. METHODS: A self administered survey was conducted in grade 10 English classes. RESULTS: The response rate was 87.1%. The prevalence of smoking was 36.2%; there was no gender difference. Only 42.8% of students ate breakfast daily; 48.8% of boys and 36.1% of girls (chi2 = 5.2; p<0.05). A higher proportion of boys (77.1%) were active for at least 30 minutes > or = 3 times/week compared to girls (66.0%) (chi2 = 4.8; p<0.05). Students who were active > or = 3 times/week were more likely to eat breakfast daily and, among boys, 60.4% of non-smokers ate breakfast daily compared to 31.9% of those currently smoking (chi2 = 13.3; p<0.001). There were no differences among girls. More girls (63.9%) were concerned about gaining weight compared to boys (36.1%) (chi2 = 37.7; p<0.001). Among girls, a higher proportion of those who were concerned about gaining weight were less likely to engage in physical activity or smoke, and more likely to skip breakfast compared to those who were not concerned. DISCUSSION: Weight concern was not associated with frequency of physical activity, smoking, or breakfast consumption among boys. The high prevalence rates for these behaviours suggests that interventions in high schools should include daily physical activity, promotion of breakfast eating (either at home or in the school), and encouragement to quit smoking. PMID- 12583679 TI - Factors influencing physical activity levels among Canadian youth. AB - OBJECTIVES: To describe the predisposing, enabling and reinforcing factors influencing the levels of physical activity among Canadian youth (ages 12-24). METHODS: Analyses of the 1996-97 National Population Health Survey (NPHS) were conducted using data collected from female (n=6195) and male (n=5925) Canadians aged 12-24 years. The data were analyzed using correlational, ANOVA and regression procedures with post hoc analyses (Bonferroni) employed where applicable. RESULTS: Compared to males, Canadian female youth were found to be less physically active, more concerned about being overweight, more depressed, and to consult mental health professionals more frequently. On the other hand, females were more likely to report greater social support than males and to be more socially involved. Those adolescents who smoke tend to be less physically active and, along with those who consume alcohol, have poorer health. DISCUSSION: The significant predisposing, enabling and reinforcing factors influencing youths' physical activity levels more profoundly affect females. The results are discussed in terms of strategies for enabling physical activity available in the literature. In addition to individual determinants influencing physical activity, a move toward understanding policy and environmental factors is recommended for further research. PMID- 12583680 TI - Intimate partner violence and health: a critique of Canadian prevalence studies. AB - OBJECTIVE: The Canadian Public Health Association, along with other professional organizations, has identified intimate partner violence (IPV) as a priority health issue to which the health professions must respond. This study synthesizes Canadian studies on the prevalence of IPV against women, focusing in particular on the stated implications for women's health and health care. METHODS: Medical and social science databases were searched for all articles pertaining to IPV in Canada for 1974 through September 2000. Reference lists of these and other related publications were consulted to supplement the literature review. Data on study characteristics, methods, and results were extracted by two independent reviewers. Discrepancies were resolved by consensus. RESULTS: Sixteen studies were identified in this review, 11 population-based and 5 conducted in clinical settings. Age, ethnicity, and socioeconomic status were not consistently documented, making comparisons and evaluations of generalizability difficult. Annual prevalence of IPV in Canada was found to range from 0.4% to 23%, with severe violence occurring from 2% to 10% annually. Less than two fifths (37.5%) of the studies incorporated a health-related measure. INTERPRETATION: This review reveals a paucity of Canadian prevalence data on IPV, marked by design and methodological issues. Poor quality data may pose a challenge to articulating and establishing a coordinated health care response to eliminating IPV in Canada. PMID- 12583681 TI - The impact of influenza-associated respiratory illnesses on hospitalizations, physician visits, emergency room visits, and mortality. AB - OBJECTIVES: Although the increased risk of hospitalization and mortality during influenza seasons has been documented extensively, there is a relative paucity of research on the impact of influenza-related illnesses on other health care use indicators, such as physician use. The purpose of this study was to examine the impact of influenza-associated respiratory illnesses on the Winnipeg health care system, including hospitalizations, physician visits and emergency room visits. Their impact on mortality was also examined. METHODS: Administrative data were used to track health care use and mortality over four influenza seasons (1995-96 to 1998-99). Excess health care use and deaths were calculated by subtracting rates during influenza seasons from those during weeks when influenza viruses were not circulating. RESULTS: Significant excess hospitalization, physician visit, and emergency room visit rates emerged for influenza and pneumonia, acute respiratory diseases, and chronic lung disease, especially among children and adults aged 65 and over. Considerable excess mortality due to influenza and pneumonia and chronic lung disease among individuals aged 65 and over also emerged, particularly among nursing home residents. DISCUSSION: Influenza associated respiratory illnesses have a substantial impact on the health care system. Given the burden of illness among children during influenza seasons, the study further suggests that influenza vaccination might be considered for this age group. PMID- 12583682 TI - Effectiveness and cost comparison of two strategies for hepatitis B vaccination of schoolchildren. AB - CONTEXT: In 1994, immunization against hepatitis B was implemented in schools in Quebec, targeting grade 4 students. In 1996-1997 and 1997-1998, one Local Community Service Centre (CLSC) replaced the school-based program in its district with vaccination offered in community clinics after school hours. The aim of the current study was to compare the effectiveness and costs of school-based and clinic-based programs. METHODS: Vaccination coverage data were collected in the CLSC with the clinic-based program (CBP), and in three matched CLSCs with a school-based program (SBP), from 1994 to 2000. Surveys were conducted to estimate costs to parents, to schools and to CLSCs in 1997-1998. RESULTS: With the implementation of the CBP, the vaccination coverage fell to 73%, compared with over 90% in the SBPs. Coverage increased to 90% when the CBP was abandoned. Costs to the CLSC were not much lower in the CBP. Societal costs were $63 per student vaccinated in the CBP, and < or = $40 in the SBPs. CONCLUSION: Results demonstrate the advantage of a SBP over a CBP for the immunization of schoolchildren. PMID- 12583683 TI - Facilitators and barriers to cervical cancer screening among Chinese Canadian women. AB - BACKGROUND: Chinese Canadian women have higher cervical cancer incidence, and lower Pap testing, rates than the general Canadian population. Predisposing, enabling and reinforcing factors associated with ever having a Pap test, and having a recent Pap test within the last 2 years, were assessed in Chinese women in British Columbia using the PRECEDE-PROCEED model. METHOD: Chinese women (n=512) between the ages of 20 and 79 years and residing in Greater Vancouver were interviewed about Pap testing, health care, traditional health beliefs, acculturation and sociodemographic characteristics. Two analyses were done, comparing women who had ever and never had a Pap test, and comparing women who had and had not received a recent Pap test. Focus groups and qualitative interviews ensured cultural sensitivity in the survey questionnaire. RESULTS: Seventy-six percent reported ever having a Pap test and 57% reported having a Pap test within the last 2 years. Traditional health beliefs were not associated with ever or recent Pap testing. However, belief that Pap testing prevented cancer and general knowledge about the Pap test were associated with screening. Concern about pain/discomfort with the test, availability of time, culturally sensitive health care services and recommendation for Pap testing by a physician were also associated with screening. Factors differed for ever, and recently, having a Pap test. INTERPRETATION: Pap testing is less common among Chinese Canadian women. Continuing education about Pap testing is recommended for physicians serving underscreened Chinese women. Culturally and linguistically appropriate educational materials are needed for the Chinese community. PMID- 12583684 TI - [Recommendations for appeal to another resource in a health case brought by nurses in the Info-Health Service CLSC: convergence between utilization declarations and informants' data]. AB - OBJECTIVES: To examine the perception of telephone advice-line users as to whether or not a formal recommendation had been made to seek another consultation and to compare users' perception to what the nurse documented. To analyze the effects of different users' and call characteristics on the incorrectness of the self-report. DATA SOURCES/STUDY SETTING: This study is a secondary analysis of data obtained from 4,696 randomly selected participants in a survey conducted among users of Info-Sante CLSC, a free-of-charge telenursing health-line service (THLS) available throughout the province of Quebec. STUDY DESIGN/DATA COLLECTION: Self-reported advice from follow-up survey phone interviews, conducted within 48 120 hours after the participant's call, were compared to the data consigned by the nurse in the computerized call-record. Covariables concerned characteristics of callers, context of the call, and satisfaction with the nurses' intervention. Association between these variables and inaccurate reports was identified using multinomial logistic regression analyses. PRINCIPAL FINDINGS: Advice to consult another health resource was recorded by the nurse in 42% of cases, whereas 39% of callers stated they had received such a recommendation. Overall disagreement between the two sources is 27% (12% by false positive and 15% by false negative) and kappa is 0.45. Characteristics such as living alone (adjusted OR = 2.5), calls relating to psychological problems (OR = 2.8), perceived seriousness (OR = 2.6) as well as others, were associated with inaccurate reports. CONCLUSIONS: Telephone health-line providers should be aware that many callers appear to interpret advice to seek additional health care differently than intended. Our findings suggest the need for continuing quality control interventions to reduce miscommunication, ensure better understanding of advice by callers, and contribute to more effective service. PMID- 12583685 TI - Drug imprinting ('unit labeling')--an update and a plea! PMID- 12583686 TI - Effects of malathion plus superphosphate or urea on Najdi sheep. AB - The effects of p.o. administration of the combination of malathion + superphosphate or urea on Najdi sheep were evaluated in sheep dosed as untreated controls, malathion-treated at 25 mg/kg/d, superphosphate-treated at 450 mg/kg/d, urea-treated at 450 mg/kg/d, malathion-treated at 25 mg/kg/d + superphosphate treated at 450 mg/kg/d, or malathion treated at 25 mg/kg/d + urea treated at 450 mg/kg/d. Oral doses of malathion alone were lethal after 6 d, and malathion + urea were fatal after 6-8 d. Malathion + superphosphate caused death after 2-3 d. Malathion, but not superphosphate or urea, inhibited serum cholinesterase activity. Hepatonephropathy correlated with changes in serum AST, ALP, cholesterol, triglyceride, bilirubin, urea, total protein and albumin. Neither malathion nor its combination with superphosphate or urea caused peripheral neuropathy. PMID- 12583687 TI - Modulation of cytochrome P450 metabolism by ergonovine and dihydroergotamine. AB - This study investigated modulation of the cytochrome P450 3A (CYP3A4)-mediated metabolism of ergotamine (ET) by ergonovine and dihydroergotamine. Liver microsomes were prepared from rats treated i.p. for 4 d with: low (10 mM) or high (100 mM) levels of dexamethasone (DM10 and DM100), dihydroergotamine, ergonovine, or control. Cytochrome P450 activity was evaluated using ET and its isomers as substrate. Ergotamine was converted to its metabolites at rates of 0.385 or 0.535 (SE = 0.040) nM/microg protein/min when incubated with liver microsomes from DM10 or DM100 treated rats, respectively. These rates were higher than for rats on other treatments. Induction of CYP34A activity was not greater for ergonovine or dihydroergotamine treatments than for controls. Both ergonovine and dihydroergotamine treatments inhibited in vitro CYP3A4 activity in a dose dependent manner producing quadratic inhibition curves. PMID- 12583688 TI - Histological alterations in the internal organs of growing chicks from feeding raw jackbean or limabean seeds. AB - The effect of raw jackbean (Canavalia ensiformis) or limabean (Phaseolus lunatus) seeds at 0, 250 or 500 g/kg in broiler chick diets for 28 d on performance and cytopathological changes in internal organs was evaluated. Relative pancreas weight increased significantly (P < 0.05) and the lungs had severe atelectasis, thickening of the alveolar septa and bronchiolar epithelial hyperplasia. Slight hemorrhages were in the intestinal walls of growing chicks fed 250 g/kg or 500 g/kg of jackbean and limabean, respectively. The livers had marked congestion of the sinusoids and centrolobular veins, while the kidneys had distension of the capillary vessels with numerous thrombi. The structural alterations in the internal organs of chicks fed the plant seeds were attributed to allelochemicals in the seeds. PMID- 12583689 TI - Blockade of sulfur mustard cytotoxicity in human epidermal keratinocytes with the purinergic receptor antagonist suramin. AB - Studies were conducted to determine whether sulfur mustard (HD)-induced cytotoxicity in human epidermal keratinocytes (HEK) could be prevented by the P2 purinergic receptor antagonist suramin. Alamar blue and calcein-AM cytotoxicity assays were conducted 48 h after HD exposure. Short (10-15 min) pre-incubations with 1-5 mM suramin blocked the cytotoxic effect of HD over a narrow HD concentration range (100-150 microM). This protective effect was suramin concentration dependent and was present if the medium was removed when HD was nearly completely hydrolyzed (60 min post-administration). Thus, suramin was required only during the actual presence of HD to exert its protective effect and not during later events required for the expression of HD-induced cytotoxicity (HD-induced cell death in HEK requires 48-72 h). These results are consistent with the hypothesis that purinergic receptors are involved in the cytotoxic effect of HD. Although suramin possesses a variety of actions unrelated to purinergic receptors, other evidence suggests that HD and suramin interact at the level of P2X receptors. Cysteine residues are primary targets of both HD and suramin. P2X receptors contain a large, cysteine-rich extracellular loop that appears critical for receptor function. PMID- 12583690 TI - Structure-activity relationships among zearalenone and its derivatives based on bovine neutrophil chemiluminescence. AB - We compared the effects of zearalenone (ZEN), an estrogenic mycotoxin produced by Fusarium fungi, and its derivatives--a- and b-zearalenols (Zel), zearalanone (ZAN), and a- and b-zearalanols (Zal)--on bovine neutrophils in vitro by using chemiluminescence, a bactericidal parameter. ZEN, a-Zel, and b-Zel suppressed luminol-dependent, phorbol 12-myristate 13-acetate-elicited chemiluminescence at a concentration of 10(-5) M, whereas ZAN, a-Zal and b-Zal did not. The suppressive zearalenols are derived from ZEN through reduction of the C6'-ketone into hydroxide, whereas the non-suppressive ZAN and Zal group possesses a hydrogenated C1'-2' bond in place of the double bond adopted in the macrolide ring or ZEN and the zearalenols. In consideration of these structure-activity relationships among ZEN and its derivatives, we conclude that possession of the C1'-2' double bond is essential for zearalenones to induce neutrophil suppression. PMID- 12583691 TI - Effects of Lantana camara (Verbenaceae) on rat fertility. AB - Lantana camara, widely used in folk medicine, possesses several pharmacological properties, including antipyretic, antimicrobial and antimutagenic properties. Lantana poisoning causes livestock mortality and morbidity; and also has adverse effects on humans working in lantana-infested forests, pastures or orchards. We examined the effects of a hydroalcoholic extract from Lantana camara var aculeata leaves on fertility of male rats. The extract did not interfere with overall weight or internal organ weights, but interfered with sperm count, daily sperm production and sperm morphology in a dose-dependent manner. PMID- 12583692 TI - Ovine metabolism of diosgenin coated on cellulose in relation to hepatogenous photosensitization of ruminants. AB - Diosgenin (2.25 g) was coated onto cellulose and dosed daily to a lamb for 12 d by intraruminal intubation. Upon completion of dosing, samples of gut contents and internal organs were subjected to chemical analysis and histological examination. No clinical signs, changes in serum chemistry, or microscopic organ changes were observed. GC-MS analyses showed that in the rumen diosgenin was metabolised to free and, to a lesser extent, conjugated derivatives of tigogenin, smilagenin and epismilagenin. Reduction of diosgenin to smilagenin and tigogenin, and epimerization of smilagenin to epismilagenin, appeared to continue in the duodenum, ileum and jejunum. Deconjugation and the epimerization of smilagenin to epismilagenin appeared to continue in the cecum and colon. Epismilagenin was the dominant conjugated sapogenin constituent identified in a bile sample. PMID- 12583693 TI - Inhalational methanol toxicity in pregnancy treated twice with fomepizole. AB - We describe a pregnant patient with a long history of inhalant abuse who came to our hospital twice during her pregnancy. In both admissions, she was treated with fomepizol (4-MP), a newly available alcohol dehydrogenase antagonist for treatment of ethylene glycol poisoning. PMID- 12583694 TI - Acute selenium poisoning from selenium-containing iron supplement in suckling pigs. AB - We report an outbreak of acute selenium poisoning among suckling pigs; 92 piglets were found dead or moribund without preceding symptoms. Necropsy revealed acute congestion of liver and small intestine. The source was a powdered iron supplement contaminated by sodium selenite. PMID- 12583695 TI - Beneficial late administration of obidoxime in malathion poisoning. AB - Early treatment of organophosphate (OP) poisoning with oximes results in reactivation of acetylcholinesterase and patient recovery. Data on efficacy of late administration of oximes, particularly obidoxime, is limited. A 42-y old woman swallowed 60 ml of 50% malathion in a suicide attempt. Characteristic muscarinic, nicotinic and central manifestations of OP poisoning appeared: atropine and 250 mg obidoxime i.v., resulted in marked improvement. Several hours after the last dose, clinical manifestations recurred and ventilation was required. After 10 d cholinesterase was still low and liver enzymes were elevated. Obidoxime was reinstituted after the 9 d interruption and muscle strength improved with the first dose. The patient could be disconnected from the ventilator and within <24 h was extubated. Oxime therapy should be considered even late in the course of untreated or partially treated OP intoxications, especially when the etiologic agent is a lipid-soluble compound (ie malathion) that can cause a protracted course of poisoning. The clinical course of this patient did not support a cause-and-effect relationship between obidoxime and the abnormal liver function. PMID- 12583696 TI - Ethylene glycol toxicosis in chickens. AB - Ethylene glycol (EG) toxicosis was diagnosed on postmortem examination in a white leghorn chicken. The clinical signs were sudden onset of depression, ataxia, convulsions and death. Natural cases of EG toxicosis have not been reported in chickens. This report highlights the importance of including EG toxicosis on differential diagnoses lists for sudden onset of depression, ataxia and death in chickens. PMID- 12583697 TI - A retrospective study of daylily toxicosis in cats. AB - The Easter, Japanese, stargazer and tiger lilies (Lilium sp) are nephrotoxic to cats. This study examined risks posed to cats by the common daylily (Hemerocallis sp: H. dumortierei, H. fulvi, H. graminea, H. seiboldii) following ingestion. Records describing ingestion of Hemerocallis sp between January 1998 and June 2002 were reviewed for signalment, quantity ingested, clinical signs (onset, severity, duration), treatments administered, and outcome. Twenty-two cases of confirmed exposure resulting in toxicosis were evaluated. Cats that ingest daylilies are at risk for gastrointestinal distress and acute renal failure. Successful treatment can be accomplished with early decontamination and aggressive fluid therapy. PMID- 12583698 TI - Pralidoxime hydrolysis of thiocholine esters. AB - Pralidoxime (2-PAM) hydrolyzes both acetylthiocholine and butytylthiocholine in an apparent first-order manner, with higher rates observed at pH 8.0 compared to those at pH 7.4. When the reaction is followed by the decrease in pH due to hydrolysis, pralidoxime hydrolyzes acetylthiocholine but not acetylcholine or succinylcholine. This implies that, while pralidoxime acts to reverse intoxication by organophosphate compounds due to the otherwise irreversible inhibition of acetylcholinesterase, it does not also supplement this detoxification by hydrolysis of the enzyme's substrate, acetylcholine. PMID- 12583699 TI - A community exposure to 2-chloro-6-fluorophenol. AB - A rural town in western New York was the site of a release of a mixture of 2 chloro-6-fluorophenol (CFP), toluene and water due to a pressure build-up at a nearby chemical facility. The regional poison control center received calls from physician offices and individual patients describing symptoms felt related to this exposure. Symptoms included headache, dry throat, cough, chest discomfort, nausea/vomiting, and diarrhea. Environmental sampling by the state health department confirmed soil and surface contamination; however, despite a noticeable odor during sampling, staff did not detect CFP in air samples. To our knowledge, although many individuals visited their primary care providers, none were hospitalized. This incident suggests that acute exposure to CFP is similar to other phenol exposures with relatively minor symptoms including headache, irritation of mucous membranes, and gastrointestinal symptoms. PMID- 12583700 TI - Poisoning of mice by Baccharis coridifolia: an experimental model. AB - The poisonous plant Baccharis coridifolia causes necrosis in lymphoid tissues and the gastrointestinal tract of cattle, horses, sheep and rabbits. An experimental poisoning of mice was undertaken to establish an experimental model in a laboratory animal specie. A single 5 to 8-g/kg dose of a suspension of the plant was administered by gavage to 11 mice. To 3 other control mice, the same volume of water was administered. Plant-dosed mice manifested clinical effects after 12 h: tachipnea, trembles, dehydration and prostration. Most of the dosed mice died 14 to 33 h after plant administration--3 survived for 12 d. Six mice had remarkable necrosis of the germinative center of secondary follicles in lymph nodes and spleen; 3 mice had necrosis of lymphoid tissues in intestine and thymus. Mice reproduce most of the lesions observed in naturally poisoned cattle and the use of this specie as an experimental model is valid. PMID- 12583701 TI - Thirty-five cases of S-carboxymethylcysteine use in paraquat poisoning. AB - The herbicide paraquat is associated with a high mortality rate. It produces multiorgan damage through the induction of acute oxidative stress, by generation of reactive oxygen species that cause oxidative damage to biomolecules. In addition to general supportive measures, the management of paraquat poisoning includes gastric washing, forced diuresis, haemodialysis and the use of antioxidants, such as N-acetylcysteine. However, this drug is rather unavailable in Venezuela and S-carboxymethylcysteine has been used. We report 35 patients with mild to severe paraquat poisoning, which beside standard supportive treatment received 1500 mg S-carboxymethylcysteine, up to 2-3 w. The mortality rate was 22.86% (8 deaths/35 cases) and was related to the severity of paraquat poisoning (as assessed by urine dithionite tests). We conclude that S carboxymethylcysteine is a reliable alternative in managing patients with paraquat poisoning. PMID- 12583702 TI - Intentional poisoning of animals in southeastern Spain: a review of the veterinary toxicology service from Murcia, Spain. AB - Data of toxicological analyses carried out over a 10-y period for suspected cases of wild and domestic animal poisonings are summarized. Of the 123 cases suspected as deliberate, 102 were be analyzed and 50 of them were positive to intentional poisoning, a total of 107 dead animals. Pesticides, especially insecticides (72%) and rodenticides (26%), were frequently involved. Aldicarb (n=15), anticoagulant rodenticides (n=8) and strychnine (n=4) were the most common toxins in baits prepared for intentional poisonings; carbofuran, methomyl, endosulfan and paraquat were also used. A coordinated Veterinary Toxicology Services network of collaboration should be set up in Spain to improve the quality of the services provided. PMID- 12583703 TI - Pesticide poisonings reported to the drug and poison information center in Izmir, Turkey. AB - This study analyzed poisonings caused by pesticides that were reported to Drug and Poison Information Center (DPIC), in Izmir from 1993 to 2001. Patient demographics, type of the pesticide, distribution according to month and year, route and reason for exposure, clinical effects and outcome were analyzed from 25,572 poisoning calls. Pesticide intoxications accounted for 8.8% of the poisonings, with 80.3% insecticides and 19.7% rodenticides. The majority of poisonings ranged from 0 to 6 y (28.2%) and 19-29 y (23.2%). Half the accidental exposures (57.7%) were in the 0-6-y group; the attempted suicide was predominant in the 19-29 y group (39.8%). Most were intoxicated with organophosphates (47.6%); 54.1% did not develop signs and symptoms of toxicity. Fatality due to pesticide poisoning was 0.4%. Preventive education against pesticide poisoning is needed. PMID- 12583704 TI - Should we test your 'hard drive' or your 'search engine'? PMID- 12583706 TI - Transformation of Bacillus subtilis in chocolate milk: evidence for low frequency of establishment of cells transformed under non-selective conditions. AB - Transformation of naturally competent Bacillus subtilis with plasmid was carried out in chocolate milk without antibiotics. Transformed cells were enumerated during the entire growth phase in chocolate milk. When DNA was added to aliquots of a batch culture after different times of incubation, transformation events were detected at all different growth stages. When DNA was added to a batch culture together with the inoculum, transformed cells were detected at the onset of exponential growth. However, apparently no or only limited growth of these transformed cells was observed. To clarify, whether the limitation of growth was due to suppression by non-transformed cells, different proportions of B. subtilis cells either carrying or not carrying the plasmid were mixed and inoculated into chocolate milk without antibiotic. Our results indicate that suppression appears to be of minor importance. Instead, plasmid-bearing cells appear to suffer from a prolonged lag-phase. However, the failure to exhibit significant growth of cells which had taken up the plasmid in chocolate milk appears to be due to failure of these cells to establish themselves as permanently transformed under non selective conditions. PMID- 12583705 TI - Construction of a marker rescue system in Bacillus subtilis for detection of horizontal gene transfer in food. AB - A marker rescue system based on the repair of the kanamycin resistance gene nptII was constructed for use in Gram-positive bacteria and established in Bacillus subtilis 168. Marker rescue was detected in vitro using different types of donor DNA containing intact nptII. The efficiency of marker rescue using chromosomal DNA of E. coli Sure as well as plasmids pMR2 or pSR8-30 ranged from 3.8 x 10(-8) to 1.5 x 10(-9) transformants per nptII gene. Low efficiencies of ca. 10(-12) were obtained with PCR fragments of 792 bp obtained from chromosomal DNA of E. coli Sure or DNA from a transgenic potato. B. subtilis developed competence during growth in milk and chocolate milk, and marker rescue transformation was detected with frequencies of ca. 10(-6) and 10(-8), respectively, using chromosomal DNA of E. coli Sure as donor DNA. Although the copy number of nptII genes of the plant DNA exceeded that of chromosomal E. coli DNA in the marker rescue experiments, a transfer of DNA from the transgenic plant to B. subtilis was detectable neither in vitro nor in situ. PMID- 12583707 TI - Lactic acid bacteria isolated from dairy products inhibit genotoxic effect of 4 nitroquinoline-1-oxide in SOS-chromotest. AB - Antigenotoxic activity against 4-nitroquinoline-1-oxide (4-NQO) of lactic acid bacteria isolated from commercial dairy products was studied using SOS Chromotest. The supernatants from bacteria-genotoxin co-incubations in general exhibited a strong suppression on SOS-induction produced by 4-NQO on the tester organism Escherichia coli PQ37 (sfiA::lacZ). High genotoxicity inhibition (>75%) was found for 31/67 of the examined bacteria and the maximum values of some strains within the species were as follows: Lactobacillus casei, 99.1%; L. plantarum, 93.3%; L. rhamnosus, 93.4%; L. acidophilus, 90.9%; L. delbrueckii subsp. bulgaricus, 85.7% and Bifidobacterium bifidum, 89.6%; Strains with low antigenotoxicity (5-60%) were evidenced in both L. acidophilus and L. delbrueckii subsp. bulgaricus, whereas some inactive strains were found only in L. casei and L. delbrueckii subsp. bulgaricus. Cell exposure to 100 degrees C for 15 min prevented antigenotoxicity and no effect was evidenced for cell-free spent media. The active strains survived at 0.1 mM 4-NQO exposure and generally presented some relevant functional properties, such as tolerance to bile (0.5%) or acid environment (pH 2.0) and adherence to Caco-2 enterocytes. Antigenotoxicity was always associated with modification of the 4-NQO absorbance profile. PMID- 12583708 TI - Taxonomy of the glycerol fermenting clostridia and description of Clostridium diolis sp. nov. AB - Six Clostridium strains which ferment glycerol to 1,3-propanediol were tested for their taxonomic and phylogenetic relatedness. All but one were known as C butyricum. By physiological tests, 16S rDNA sequences and fatty acid composition two groups were distinguished. The first comprised the strains VPI 3266, DSM 2478 and DSM 523 (C. "kainantoi") and was consistent with the type strain of C. butyricum in almost all characters. The second group comprising the strains DSM 5430, DSM 5431 and E5 was related to C. beijerinckii. The 16S rDNAs of these strains were almost identical with that of the type strain of C. beijerinckii, DSM 791. The DNA-DNA hybridization value of DSM 5431 and ES with C. beijerinckii DSM 791 was markedly but not decisively lower (67 and 72%, respectively). However, there were significant physiological differences to C. beijerinckii which suggested to describe the strains as a separate species, Clostridium diolis with strain SH1 (= DSM 5431) as the type strain. The new species is distinguished from C. beijerinckii, which requires complex nutrients, by its ability to grow in glucose mineral medium with biotin as the only growth factor and by differences in substrate utilization. "C. kainantoi" Takeda and Matsui was recognized as a later synonym of C. butyricum. PMID- 12583709 TI - Allisonella histaminiformans gen. nov., sp. nov. A novel bacterium that produces histamine, utilizes histidine as its sole energy source, and could play a role in bovine and equine laminitis. AB - When cattle and horses are fed large amounts of grain, histamine can accumulate in the gastrointestinal tract, and this accumulation can cause an acute inflammation of the hooves (laminitis). When ruminal fluid from dairy cattle fed grain supplements was serially diluted in anaerobic MRS medium containing histidine (50 mM), histamine was detected at dilutions as high as 10(-7). The histidine enrichments were then transferred successively in an anaerobic, carbonate-based medium (50 mM histidine) without glucose. The histamine producing bacteria could not be isolated from the rumens of cattle fed hay; however, histamine producing bacteria could be isolated the feces of cattle fed grain and the cecum of a horse. All of the histamine producing isolates had the same ovoid morphology. The cells stained Gram-negative and were resistant to the ionophore, monensin (25 microM). The doubling time was 110 min, and the yield was 1.5 mg cell protein per mmol histidine. The G+C content was 46.8%. Lysine was the only other amino acid used, but lysine did not allow growth if histidine was absent. Because carbohydrate and organic acid utilization was not detected, it appeared that the isolates used histidine decarboxylation as their sole mechanism of energy derivation. 16s rRNA gene sequencing indicated that the isolates were most closely related to low G+C Gram-positive bacteria (firmicutes), but similarities were < or = 94%. Because the most closely related bacteria (Dialister pneumonsintes, Megasphaera elsdenii and Selenomonas ruminantium) did not produce histamine from histidine, we propose that these histamine producing bacteria be assigned to a new genus, Allisonella, as Allisonella histaminiformans gen. nov., sp. nov. The type strain is MR2 (ATCC BAA610, DSM 15230). PMID- 12583710 TI - Burkholderia tuberum sp. nov. and Burkholderia phymatum sp. nov., nodulate the roots of tropical legumes. AB - The taxonomic status of five root nodule isolates from tropical legumes was determined using a polyphasic taxonomic approach. Two isolates were identified as B. caribensis, an organism originally isolated from soil in Martinique (the French West Indies). One isolate was identified as Burkholderia cepacia genomovar VI, a B. cepacia complex genomovar thus far only isolated from sputum of cystic fibrosis patients. The remaining two isolates were identified as novel Burkholderia species for which we propose the names Burkholderia tuberum sp. nov. and Burkholderia phymatum sp. nov. The type strains are LMG 21444T and LMG 21445T, respectively. PMID- 12583711 TI - Isolation and characterization of a new type of aerobic, oxalic acid utilizing bacteria, and proposal of Oxalicibacterium flavum gen. nov., sp. nov. AB - A mesophilic, aerobic oxalic acid utilizing yellow-pigmented bacterium has been isolated from litter of oxalate producing plants in the region of Izmir (Turkey). It is motile by means of 1-3 polar flagella. Optimal growth occurred between 25 30 degrees C at pH 6.9. The G+C content of DNA is 62-64 mol % (Tm). Based on its morphological and biochemical features the organism belongs to the genus Pseudomonas, but differs from all the previously described species. The taxonomic relationships among strains described as or previously tentatively assigned to the genus Pseudomonas were investigated using numerical classification, DNA base composition and DNA-DNA hybridization. 16S rDNA sequences were determined for the strain TA17. On the basis of 16S rDNA sequence comparisons, physiological and biochemical characteristics, it is proposed to classify TA17T in a new genus and species for which the name Oxalicibacterium flavum gen. nov., sp. nov. is proposed. The type strain is TA17T (= NEU98T, = LMG 21571T). PMID- 12583712 TI - 16S-23S rDNA intergenic spacer region polymorphism of Lactococcus garvieae, Lactococcus raffinolactis and Lactococcus lactis as revealed by PCR and nucleotide sequence analysis. AB - The intergenic spacer region (ISR) between the 16S and 23S rRNA genes was tested as a tool for differentiating lactococci commonly isolated in a dairy environment. 17 reference strains, representing 11 different species belonging to the genera Lactococcus, Streptococcus, Lactobacillus, Enterococcus and Leuconostoc, and 127 wild streptococcal strains isolated during the whole fermentation process of "Fior di Latte" cheese were analyzed. After 16S-23S rDNA ISR amplification by PCR, species or genus-specific patterns were obtained for most of the reference strains tested. Moreover, results obtained after nucleotide analysis show that the 16S-23S rDNA ISR sequences vary greatly, in size and sequence, among Lactococcus garvieae, Lactococcus raffinolactis, Lactococcus lactis as well as other streptococci from dairy environments. Because of the high degree of inter-specific polymorphism observed, 16S-23S rDNA ISR can be considered a good potential target for selecting species-specific molecular assays, such as PCR primer or probes, for a rapid and extremely reliable differentiation of dairy lactococcal isolates. PMID- 12583714 TI - Species specific identification of nine human Bifidobacterium spp. in feces. AB - Based on the 16S rDNA sequences, species specific primers were designed for the rapid identification by DNA amplification of nine human Bifidobacterium spp., namely B. adolescentis, B. angulatum, B. bifidum, B. breve, B. catenulatum, B. dentium, B. infantis, B. longum, B. pseudocatenulatum. B. lactis currently included in dairy products was added to the series. The primers were designed to target different positions of the 16S rDNA, allowing the simultaneous identification of these ten species of Bifidobacterium using two mixtures of primers. The identification procedure described in this paper was validated by establishing a correlation with an AluI restriction pattern of the different full length amplified 16S rDNA. This multiple primer DNA amplification technique was applied for the identification of pure colonies of Bifidobacterium spp. or directly from total bacteria recovered from human fecal samples. The technique was shown to be useful to detect dominant species and, when primers were used in separate reactions, underrepresented species could be identified as well. PMID- 12583713 TI - Use of 16S-23S rRNA spacer-region (SR)-PCR for identification of intestinal clostridia. AB - The suitability of a species identification technique based on PCR analysis of 16S-23S rRNA spacer region (SR) polymorphism for human intestinal Clostridium species was evaluated. This SR-PCR based technique is highly reproducible and successfully differentiated the strains tested, which included 17 ATCC type strains of Clostridium and 152 human stool Clostridium isolates, at the species or intraspecies level. Ninety-eight of 152 stool isolates, including C. bifermentans, C. butyricum, C. cadaveris, C. orbiscindens, C. paraputrificum, C. pefringens, C. ramosum, C. scindens, C. spiroforme, C. symbiosum and C. tertium, were identified to species level by SR-PCR patterns that were identical to those of their corresponding ATCC type strains. The other 54 stool isolates distributed among ten SR-PCR patterns that are unique and possibly represent ten novel Clostridium species or subspecies. The species identification obtained by SR-PCR pattern analysis completely agreed with that obtained by 16S rRNA sequencing, and led to identification that clearly differed from that obtained by cellular fatty acid analysis for 23/152 strains (15%). These results indicate that SR-PCR provides an accurate and rapid molecular method for the identification of human intestinal Clostridium species. PMID- 12583716 TI - Detection of 16S rDNA sequences representing the novel phylum "Nanoarchaeota": indication for a wide distribution in high temperature biotopes. AB - We screened samples from high temperature biotopes for 16S rRNA genes of the novel archaeal phylum "Nanoarchaeota". Positive PCR amplifications were obtained from Yellowstone National Park, Uzon Caldera, and an abyssal vent system. These sequences form a cluster with the sequence of "Nanoarchaeum equitans", indicating a wide distribution of this phylum. PMID- 12583717 TI - Improvement of ciliate identification and quantification: a new protocol for fluorescence in situ hybridization (FISH) in combination with silver stain techniques. AB - A new protocol for taxon specific probe based fluorescent in situ hybridization was developed for the identification and quantification of ciliates in microbial communities. Various fixatives and experimental parameters were evaluated and optimized with respect to cell permeability and morphological preservation. Optimum results were adaption by obatined of a modified fixation method using Bouin's solution. Furthermore, conventional staining procedures such as different Protargol stain techniques and a silver nitrate impregnation method were modified and can now be applied in combination with fluorescence in situ hybridization. The new protocol allows a rapid and reliable identification as well as quantification of ciliates based upon classical morphological aspects and rRNA based phylogenetic relationships performed in one experiment. Furthermore, a set of specific probes targeting different regions of the 18S rRNA was designed for Glaucoma scintillans Ehrenberg, 1830 and tested by applying this new approach of combining in situ cell hybridization with conventional staining techniques. PMID- 12583715 TI - Comparison of ribotyping, randomly amplified polymorphic DNA, and pulsed-field gel electrophoresis for molecular typing of Vibrio tapetis. AB - Brown ring disease, caused by Vibrio tapetis, is an important pathological problem in different species of cultured clams. In order to evaluate the genetic diversity of the pathogen, twenty-seven isolates of V tapetis with different origin were screened by ribotyping (RT), pulsed-field gel electrophoresis (PFGE) and randomly amplified polymorphic DNA PCR (RAPD). Restriction with PvuII, SalI, and SmaI gave 2 RT patterns, differentiating in all cases the strain 0202RD isolated from carpet-shell clams (Ruditapes decussatus) from the other strains tested. The use of NotI generated strain specific PFGE profiles, which could be grouped in two main clusters. Cluster 1 grouped all but one strain and was subdivided into six PFGE subtypes (1a to 1f) which joined at a similarity level of 75.6%. Cluster 2 included again only the isolate 0202RD. RAPD analysis yielded the same results with three different primers, this method being able to differentiate the isolates from R. decussatus from those isolated from other clam species. Of the three techniques evaluated, PFGE was the most discriminating of the three techniques evaluated, followed in discriminating power by RAPD and RT tests. On the basis of the results obtained, we conclude that the RAPD procedure, which is more rapid and easier to perform than the other techniques, shows to be very useful to analyze large amounts of strain collections from an epidemiological monitoring stanpoint. In addition, PFGE is of great utility to evaluate the genetic diversity of strains involved in an outbreak and to study the spreading of a specific clone. PMID- 12583719 TI - Cellulose-degrading potentials and phylogenetic classification of carboxymethyl cellulose decomposing bacteria isolated from soil. AB - In a previous study, culturable carboxymethyl-cellulose (CMC) decomposing soil bacteria isolated from different sampling positions across an agricultural encatchment have been classified into 31 pattern groups by digestion of amplified 16S rDNA using a single restriction enzyme (Ulrich and Wirth: Microb. Ecol. 37, 238-247, 1999). In order to reveal relationships between phylogenetic diversity and phenotypic functions, a further differentiation of two selected site-specific pattern groups (I and H) was performed, resulting in a sub-classification of four and three ARDRA groups, respectively. Based on sequencing a representative isolate of each ARDRA group, the isolates were assigned to the genus Streptomyces. The ARDRA groups were dispersed across various clades of the genus with a direct affiliation to species known for cellulolytic activity in one group, only. The isolates differed in potentials to degrade colloidal, native or highly crystalline cellulose derivatives. Out of 39 isolates, 11 were capable of degrading all substrates, 17 were restricted to degrade CMC only, and 11 were active decomposers of exclusively both CMC and colloidal cellulose. In most cases, the genetic classification of the isolates corresponded with groupings based on cellulose degrading capabilities. Thus, isolates of four ARDRA groups were restricted to the degradation of CMC, while two further isolates which efficiently degraded all cellulose derivatives formed two separate ARDRA groups. The major ARDRA group, however; displayed a high variability of degradation capabilities. The study of additional phenotypic features revealed a broad potential to decompose a set of various carbon substrates, which matched the phylogenetic classification in several cases. PMID- 12583718 TI - Phenotypic and genotypic diversity of rhizobia nodulating Pterocarpus erinaceus and P. lucens in Senegal. AB - A total of fifty root nodules isolates of fast-growing and slow growing rhizobia from Pterocarpus ennaceus and Pterocarpus lucens respectively native of sudanean and sahelian regions of Senegal were characterized. These isolates were compared to representative strains of known rhizobial species. Twenty-two new isolates were slow growers and twenty-eight were fast growers. A polyphasic approach was performed including comparative total protein sodium dodecyl sulphate polyacrylamide gel (SDS-PAGE) profile analysis; 16S rDNA and 16S-23S rDNA intergenic spacer (IGS) sequence analysis. By SDS-PAGE the slow growing isolates grouped in one major cluster containing reference strains of Bradyrhizobium sp. including strains isolated in Africa, in Brazil and in New Zealand. Most of the fast-growing rhizobia grouped in four different clusters or were separate strains related to Rhizobium and Mesorhizobium strains. The 16S rDNA and 16S-23S rDNA IGS sequences analysis showed accurately the differentiation of fast growing rhizobia among the Rhizobium and Mesorbizobium genospecies. The representative strains of slow growing rhizobia were identified as closely related to Bradyrbizobium elkanii and Bradyrhizobium japonicum. Based on 16S rDNA sequence analysis, one slow growing strain (ORS199) was phylogenetically related to Bradyrbizobium sp. (Lupinus) and Blastobacter denitrificans. This position of ORS 199 was not confirmed by IGS sequence divergence. We found no clear relation between the diversity of strains, the host plants and the ecogeographical origins. PMID- 12583720 TI - Soils of the Chinese Hubei province show a very high diversity of Sinorhizobium fredii strains. AB - Biodiversity studies of native soybean-nodulating rhizobia in soils from the Chinese Hubei province (Honghu county; pH 8, alluvial soil) have been carried out. Inoculation of an American (Williams) and an Asiatic (Peking) soybean cultivar with eleven soil samples led to the isolation of 167 rhizobia strains. The ratio (%) of slow-/fast-growing isolates was different depending on the trap plant used. All isolates were able to nodulate both cultivars, although the N2 fixation efficiency (measured as plant-top dry weight) was different among them. A total of thirty-three isolates were selected for further characterisation on the basis of physiological parameters, PCR-RFLP of symbiotic genes and Low Molecular Weight RNA, lipopolysaccharide, protein and plasmid profiles. Low Molecular Weight RNA profiling indicates that all the isolates belong to species Sinorhizobium fredii. The dendrogram obtained with the physiological parameters has been useful to classify the isolates at strain level, although plasmid profiling was the most discriminating technique to detect differences among the analysed soybean-rhizobia isolates, showing there is not two isolates identical each other. Plasmid profile analyses also revealed that some of the investigated strains contain low molecular weight plasmids (7-8-kb). They are, to our knowledge, the smallest ever found in rhizobia and they could be the starting point for the construction of the first group of vectors based on a native rhizobia replicon. PMID- 12583721 TI - Diversity of 746 heterotrophic bacteria isolated from microbial mats from ten Antarctic lakes. AB - Microbial mats, growing in Antarctic lakes constitute unique and very diverse habitats. In these mats microorganisms are confronted with extreme life conditions. We isolated 746 bacterial strains from mats collected from ten lakes in the Dry Valleys (lakes Hoare and Fryxell), the Vestfold Hills (lakes Ace, Druzhby, Grace, Highway, Pendant, Organic and Watts) and the Larsemann Hills (lake Reid), using heterotrophic growth conditions. These strains were investigated by fatty acid analysis, and by numerical analysis, 41 clusters, containing 2 to 77 strains, could be delineated, whereas 31 strains formed single branches. Several fatty acid groups consisted of strains from different lakes from the same region, or from different regions. The 16S rRNA genes from 40 strains, representing 35 different fatty acid groups were sequenced. The strains belonged to the alpha, beta and gamma subclasses of the Proteobacteria, the high and low percent G+C Gram-positives, and to the Cytophaga-Flavobacterium Bacteroides branch. For strains representing 16 fatty acid clusters, validly named nearest phylogenetic neighbours showed pairwise sequence similarities of less than 97%. This indicates that the clusters they represent, belong to taxa that have not been sequenced yet or as yet unnamed new taxa, related to Alteromonas, Bacillus, Clavibacter, Cyclobacterium, Flavobacterium, Marinobacter, Mesorhizobium, Microbacterium, Pseudomonas, Saligentibacter, Sphingomonas and Sulfitobacter. PMID- 12583722 TI - Study of the bacterial load in a gelatine production process focussed on Bacillus and related endosporeforming genera. AB - Gelatine is an animal protein with many industrial applications. Previous studies pointed out that endosporeforming bacteria, belonging to the genus Bacillus or related genera, might contaminate and survive the production process of gelatine, leading to products of low quality and safety. The aim of this study is to determine the bacterial diversity of contaminants isolated from a gelatine production chain with emphasis on aerobic endosporeforming bacteria. Contaminants were isolated from samples taken at five crucial points along two different production lines of a gelatine production process and from water supplies used for extraction and cooling. Gaschromatographic methyl ester analysis of fatty acids was performed to differentiate isolates at the genus level. Apart from members of the genus Bacillus or related endosporeforming genera, also members of Salmonella, Kluyvera, Staphylococcus, Burkholderia, Enterococcus, Pseudomonas, Yersinia, Streptococcus and Brevundimonas could be detected. Isolates identified as belonging to Bacillus and related endosporeforming genera were further characterised by gelatinase tests, rep-PCR and 16S rDNA sequencing. All these isolates showed the ability to liquefy gelatine. Endosporeforming isolates were assigned to Bacillus licheniformis, B. fumarioli, members of the B. cereus group, B. badius, B. coagulans, B. subtilis, Brevibacillus agri, Alicyclobacillus acidocaldarius and a yet undescribed Paenibacillus species. PMID- 12583723 TI - Enumeration of thermophilic Bacillus species in composts and identification with a Random Amplification Polymorphic DNA (RAPD) protocol. AB - The thermophilic microbial flora of general garden and domestic wastes composts, derived from thermogenic, post-thermogenic and maturation phases, was analysed using spore and total plate counts in combination with an optimised RAPD protocol. A total of 459 isolates were recovered obtained at 55 degrees C, and another 56 at 70 degrees C using tryptic soy-starch agar plates, with near-equal numbers being derived from total plate counts or spore preparations. The isolates were obtained from 11 compost samples and were assigned to eighteen different RAPD fingerprint types, with 76.1% of these ultimately being positively assigned by their RAPD profiles to just 2 species including Bacillus thermodenitrificans and B. licheniformis. Viable cell numbers ranged from 1.4 to 150 x 10(6) colony forming units per gram compost (wet weight), with the highest two counts being from 2 week and 4 week old compost samples with temperatures of 70 degrees C and 55 degrees C, respectively. B. thermodenitrificans was a dominant isolate (representing more than 50% of isolates from total plate counts) in 7 of the 11 individual compost total plate count samples between 30 degrees C to 73 degrees C, and accounted for 68.9% of all isolates overall. Another relatively common Bacillus species that was identified with RAPDs in significant numbers included B. licheniformis (7.2% of all isolates and dominant isolate in 1 sample). Three other relatively common RAPD profiles could not be identified by comparison with known species in a RAPD profile database but were tentatively identified using 16S rDNA sequence comparisons. These were B. sporothermodurans (4.9% of all isolates and dominant in 1 sample), B. thermosphaericus (7.4% and dominant in 1 sample) and Terrabacter tumescens (5.0%). Overall, based on the vegetative and spore count results and the subsequent RAPD-based identification, the data strongly support a significant role for B. thermodenitrificans in the composting process, and casts doubt on the notion that B. stearothermophilus sensu strictu (DSMZ 22) is a prominent member within compost ecology. PMID- 12583724 TI - Synthesis of nucleoside dimers bridged on ribose with a butadiynyl group. AB - [reaction: see text] The nucleoside dimer linked by a butadiynediyl group at C 3'beta may serve as a building block for the preparation of backbone-modified oligonucleotides for DNA repair or mutation in functional genomics. We prepared this type of dimer by an Eglington or Sonogashira coupling reaction. The unsymmetrical dimer was synthesized by coupling the acetylene with the bromoacetylene. Only marginal cytotoxicity was detected for one of the dimers. PMID- 12583725 TI - Role of hyperconjugation in determining carbon-oxygen bond dissociation enthalpies in alkylperoxyl radicals. AB - [reaction: see text] Theoretical calculations of carbon-oxygen bond dissociation enthalpies in substituted methylperoxyl radicals (YCH(2)OO*) reveal that bond strengths are not governed solely by the stability of YCH2* radicals but are largely affected by hyperconjugation when Y is electron-donating or conjugating. In many cases, this hyperconjugative effect is greater than stabilization of the methyl radical by Y. All electron-withdrawing Y exert small destabilizing effects via inductive withdrawal of electrons from the polarized C-OO* bond. PMID- 12583726 TI - Facile construction and divergent transformation of polycyclic isoxazoles: direct access to polyketide architectures. AB - [reaction: see text] Base-promoted cyclocondensation of C-chloro oximes with cyclic 1,3-diketones affords functionalized isoxazoles in good yield and under convenient reaction conditions. This process enables the synthesis of highly substituted products with notable functional group tolerance. The products obtained are directly converted to a variety of polyketide-derived polycyclic structures including xanthenes, anthracenes, and benzophenones. PMID- 12583727 TI - Isoxazole --> benzisoxazole rearrangement promoted cascade reactions affording stereodefined polycycles. AB - [reaction: see text] A new chemo-, regio-, and stereoselective cascade reaction features a novel isoxazole --> benzisoxazole rearrangement and affords highly functionalized, differentially protected compounds. The products of this reaction are directly converted to a number of complex, structurally diverse polycyclic molecules. These transformations highlight the unique chemistry inherent to readily prepared fused isoxazoles. PMID- 12583728 TI - Unusual, strained heterocycles: 3-alkylidene-2-methyleneoxetanes from Morita Baylis-Hillman-type adducts. AB - [reaction: see text] 3-Alkylidene-2-methyleneoxetanes have been prepared by treating alpha-alkylidene-beta-lactones, derived from Morita-Baylis-Hillman-type adducts, with dimethyltitanocene. Preliminary studies of the reactivity of these little known, strained heterocycles are also described. PMID- 12583729 TI - 2'-O-[2-(amino)-2-oxoethyl] oligonucleotides. AB - [structure: see text] Oligonucleotides with novel modifications, 2'-O-[2-(amino) 2-oxoethyl] (2'-O-NAc), 2'-O-[2-(methylamino)-2-oxoethyl] (2'-O-NMAc), 2'-O-[2 (dimethylamino)-2-oxoethyl] (2'-O-DMAc), and 2'-O-[2-[[2 (dimethylamino)ethyl]amino]-2-oxoethyl] (2'-O-DMAEAc), have been synthesized. These modified oligonucleotides exhibit high binding affinity to complementary RNA (and not to DNA) and considerably enhance the nuclease stability of oligonucleotides with t(1/2) > 24 h. PMID- 12583730 TI - Influences of catalyst configuration and catalyst loading on selectivities in reactions of diazoacetamides. Barrier to equilibrium between diastereomeric conformations. AB - [reaction: see text] Stereoelectronic factors present a barrier to equilibrium between diastereomeric conformations resulting in differences in selectivity as a function of catalyst configuration. The bis(trimethylsilyl)-methyl protective group is inert to insertion but directs carbon-hydrogen insertion with enhanced enantiocontrol. PMID- 12583731 TI - Nonselective bromination-selective debromination strategy: selective bromination of unsymmetrical ketones on singly activated carbon against doubly activated carbon. AB - [reaction: see text] We have found a new synthetic method for the preparation of the alpha-bromoketones that are brominated in the less activated terminal position of unsymmetrical ketones. Brominations in short reaction times (kinetically controlled) provided internally brominated compounds as a major product. However, brominations in longer reaction times (thermodynamically controlled) gave more of the terminally brominated compound through the reversible reaction by Br(2) and produced hydrogen bromide. Several brominated compounds at the terminal position were successfully prepared through the new synthetic route. PMID- 12583732 TI - New stable backbone linker resins for solid-phase peptide synthesis. AB - [reaction: see text] Two new 4-methoxybenzaldehyde backbone linker resins were developed for the solid-phase synthesis of peptides. The linkers are very stable during the cleavage of common protecting groups for amines (Fmoc, Boc) and carboxylic acids (Me, All, tBu) in peptide synthesis. Cleavage from the resin with refluxing TFA is sufficiently mild for peptides containing polar and nonpolar amino acids. PMID- 12583734 TI - Highly efficient, general procedure for the preparation of alkylzinc reagents from unactivated alkyl bromides and chlorides. AB - [reaction: see text] Alkylzinc bromides have been efficiently prepared by the direct insertion of zinc metal (dust, powder, granule, shot), activated with 1-5 mol % I(2), into alkyl bromides in a polar aprotic solvent. The zinc reagents thus formed undergo Ni- and Pd-catalyzed cross-coupling with aryl halides to produce functionalized alkylarenes in excellent yields. PMID- 12583733 TI - Samarium(II) iodide mediated radical/polar crossover reactions of cyclobutenes. An efficient approach to the BCD ring system of the penitrems. AB - [reaction: see text] Radical/polar crossover reactions of derivatives of 1-(2 cyclobutenyl)-2-(2-iodoaryl)ethanones with acetone promoted by samarium diiodide and HMPA provide 1-(1-hydroxy-1-methylethyl)-2,2a,4,8b-tetrahydro-1H cyclobuta[a]naphthalen-3-one derivatives in about 50% isolated yield. This reaction shows promise for construction of the BCD ring fragment of the penitrems. PMID- 12583735 TI - C2-symmetric bissulfoximines as ligands in copper-catalyzed enantioselective Diels-Alder reactions. AB - [reaction: see text] Bissulfoximines have been used as chiral ligands in copper catalyzed enantioselective Diels-Alder reactions between acryloyl-2 oxazolidinones and cyclopentadiene. After optimizing the ligand structure, the metal source, the counterions, and the solvent, products with up to 93% ee have been obtained. PMID- 12583736 TI - Exploring the optimal site for modifications of pyranmycins with the extended arm approach. AB - [reaction: see text] Continuing from the syntheses and the antibacterial studies of a library of pyranmycins, we further probed the proximity around ring III of pyranmycin by introducing an "extended arm" that has hydroxyethyl or aminoethyl groups at the O-2' ', O-3' ', or O-4' ' positions. The results from the antibacterial studies reveal the optimal structural motif is the attachment of an extended arm with a terminal hydroxyl group at the O-3' ' position. PMID- 12583737 TI - A novel microwave-mediated one-pot synthesis of indolizines via a three-component reaction. AB - [reaction: see text] The microwave-mediated three-component reaction of acyl bromide, pyridine, and acetylene is catalyzed by basic alumina to give corresponding indolizines in excellent yields in a one-pot reaction. PMID- 12583738 TI - New palladium(II)-catalyzed asymmetric 1,2-dibromo synthesis. AB - [reaction: see text] The oxidation of olefins by chiral monometallic and bimetallic Pd(II)-Cu(II) catalysts in bromide-containing aqueous-THF reaction mixtures produced chiral 1,2-dibromides. With alpha-olefins, the ee's were about 95% while most of the internal alkenes gave somewhat lower enantioselectivities ( approximately 80%). PMID- 12583739 TI - Synthesis of new 14-membered macrolide antibiotics via a novel ring contraction metathesis. AB - [reaction: see text] A novel ring opening ring closing metathesis (ROM-RCM) was demonstrated for cyclic conjugated dienes, effecting the excision of a C(2)H(2) unit and a net ring contraction. Applying the ring contraction metathesis, new 14 membered ring macrolide antibiotics were synthesized in a single step from existing 16-membered ring macrolides. This new class of macrolide antibiotics will provide access to new therapeutics for the treatment of macrolide-resistant bacterial infections. PMID- 12583740 TI - Syntheses of both the putative and revised structures of aeruginosin EI461 bearing a new bicyclic alpha-amino acid. AB - [structure: see text] The putative structure of the naturally occurring aquatic peptide aeruginosin EI461 has been prepared from d-tyrosine. A corrected structure for aeruginosin EI461 is proposed, and the structure is proven by synthesis, which was accomplished using the new alpha-amino acid (2S,3aR,6R,7aR) 6-hydroxy-2-carboxyoctahydroindole, prepared from l-tyrosine. Succesive couplings of the dipeptide d-Leu-3a,7a-diepi-l-Choi with l-Hpla and NH(4)OH and a deprotection step gave aeruginosin EI461. PMID- 12583741 TI - Synthesis of (-)-TAN-2483A. Revision of the structures and syntheses of (+/-)-FD 211 (waol A) and (+/-)-FD-212 (waol B). AB - [structure: see text] The structure of waol A has been revised from 1 to 6, the vinylogue of TAN-2483 A (5). Aldol reaction of 10b(c) with 2,4-hexadienal (11) affords 9b(c), which is subjected to iodoetherification with bis(sym collidine)IPF(6) to provide 8b(c). Treatment with Et(3)N in CH(2)Cl(2) completes three-step syntheses of TAN-2483A (5) and waol A (6). PMID- 12583742 TI - S-Benzoxazolyl (SBox) glycosides as novel, versatile glycosyl donors for stereoselective 1,2-cis glycosylation. AB - [reaction: see text] Novel glycosyl donors, S-benzoxazolyl (SBox) glycosides, have been synthesized, tested toward various protecting group manipulations, and applied to the highly stereoselective 1,2-cis glycosylation. These compounds fulfill the requirements for a modern glycosyl donor such as accessibility, high stability toward protecting group manipulations, and mild activation conditions. It was also demonstrated that SBox glycosides withstand other glycosyl donor activation conditions and therefore allow selective glycosylations of O-pentenyl and thioglycosides. PMID- 12583743 TI - Tandem cross-metathesis/hydrogenation/cyclization reactions by using compatible catalysts. AB - [reaction: see text] A one-pot tandem cross-metathesis/hydrogenation/cyclization procedure was achieved at room temperature, under 1 atm of hydrogen, in the presence of a ruthenium catalyst and PtO(2) showing the compatibility of the two catalysts. This tandem reaction allows the synthesis of substituted lactones and lactols from acrylic acid and acrolein, respectively, in the presence of unsaturated alcohols. PMID- 12583744 TI - Divergent regioselectivity in the base-promoted reactions of cyclic eight membered alpha-ketols with activated halides. AB - [reaction: see text] Deprotonation of 2-hydroxycyclooctanone followed by exposure to an allylic or benzylic halide proceeds very selectively to give the product of C-alkylation. The effect of Delta(5,6)-unsaturation is to promote instead the formation of the O-alkylated derivative. This crossover in kinetic preference is attributed to an inability of the olefinic system to attain a conformation conducive to proton abstraction at C-2. PMID- 12583745 TI - Synthesis, stereostructure, and conformations of novel bi- and trifunctional (+) isomenthone derivatives. AB - [reaction: see text] Following protection, keto alcohols 4 [from (+)-isomenthone] undergo reduction to 1,3-diol 6 (S configuration at the new stereocenter). Organometallic C-nucleophiles add to the carbonyl with the same facial selectivity as hydride, providing multifunctional derivatives, e.g., 8 and 9, with five contiguous stereocenters. The side chain hydroxyl of 4 is elaborated into amino and amide derivatives (e.g., 12). Structural analysis shows that 6, 8, 9, and the precursor to 12 (10) all adopt triaxial solid-state conformations. PMID- 12583747 TI - Mono- versus bidentate ligands in rhodium-catalyzed asymmetric hydrogenation. A comparative rate study. AB - [reaction: see text] Bidentate chiral phosphines are no longer essential for achieving a fast and highly enantioselective hydrogenation of alpha- or beta dehydroamino acid derivatives. In particular, a readily accessible and stable monodentate phosphoramidite can be highly effective in these asymmetric hydrogenations. PMID- 12583746 TI - Rational design of biologically important chemosensors: a novel receptor for selective recognition of acetylcholine over ammonium cations. AB - [structure: see text] In consideration of competition between cation-pi and hydrogen bond interaction forces, we designed a novel receptor, 1,3,5 tris(pyrrolyl)benzene, which shows high selectivity for acetylcholine (ACh). The selectivity of the receptor for ACh over other ammonium cations is demonstrated by the ion-selective electrode (ISE) method in buffer solution. The binding free energy of the receptor with ACh in chloroform solution is measured to be 3.65 kcal/mol in the presence of chloride anion by nuclear magnetic resonance spectroscopy, and that in water is estimated to be much greater ( approximately 6 kcal/mol). PMID- 12583748 TI - Asymmetric intermolecular C-H activation, using immobilized dirhodium tetrakis((S)-N-(dodecylbenzenesulfonyl)- prolinate) as a recoverable catalyst. AB - [reaction: see text] Heterogenization of dirhodium tetrakis((S)-N dodecylbenzenesulfonyl)prolinate) (Rh(2)(S-DOSP)(4)) can be readily achieved on a pyridine functionalized highly cross-linked polystyrene resin. The immobilized complex is readily recycled and exhibits excellent catalytic activity for asymmetric intermolecular C-H activation by means of rhodium carbenoid induced C H insertion. PMID- 12583749 TI - Intramolecular titanium-mediated aminocyclopropanation of terminal alkenes: easy access to various substituted azabicyclo[n.1.0]alkanes. AB - [reaction: see text] A variety of substituted azabicyclo[n.1.0]alkanes were synthesized by intramolecular titanium-mediated cyclopropanation of N-benzyl-N-(2 alkylalk-3-enyl)formamides and N-benzyl-N-alkadienylformamides. N-Benzylpyrroline upon treatment with Grignard reagents undergoes a titanium-mediated carbomagnesiation to yield N-benzyl-N-(2-alkylbut-3-enyl)amines. PMID- 12583750 TI - Toward a structure-activity relationship for apoptolidin: selective functionalization of the hydroxyl group array. AB - [reaction: see text] To investigate the structural basis for the exceptional selectivity and activity of apoptolidin (1), a strategy has been devised that allows for selective functionalization of seven of its eight hydroxyl groups based on progressive silyl protection, derivatization, and deprotection. The syntheses of these derivatives and their ability to inhibit F(0)F(1)-ATPase are reported. PMID- 12583751 TI - Synthesis of (-)-matairesinol, (-)-enterolactone, and (-)-enterodiol from the natural lignan hydroxymatairesinol. AB - [reaction: see text] We describe here a four-step semisynthetic method for the preparation of enantiomerically pure (-)-enterolactone starting from the readily available lignan hydroxymatairesinol from Norway spruce (Picea abies). Hydroxymatairesinol was first hydrogenated to matairesinol. Matairesinol was esterified to afford the matairesinyl 4,4'-bistriflate, which was deoxygenated by palladium-catalyzed reduction to 3,3'-dimethylenterolactone. Demethylation of 3,3'-dimethylenterolactone and reduction with LiAlH(4) yielded (-)-enterolactone and (-)-enterodiol, respectively. PMID- 12583752 TI - Catalytic asymmetric synthesis of both syn- and anti-3,5-dihydroxy esters: application to 1,3-polyol/alpha-pyrone natural product synthesis. AB - [reaction: see text] We describe a catalytic asymmetric synthesis of both syn- and anti-3,5-dihydroxy esters. The method relies upon catalytic asymmetric epoxidation of alpha,beta-unsaturated imidazolides and amides, using lanthanide BINOL complexes, and diastereoselective reduction of ketones. The method was applied to the enantioselective syntheses of 1,3-polyol/alpha-pyrone natural products 9a, 9b, and strictifolione (10). The absolute stereochemistry of 9a and 9b was also determined. PMID- 12583753 TI - Synthesis of a simplified bryostatin C-ring analogue that binds to the CRD2 of human PKC-alpha and construction of a novel BC-analogue by an unusual Julia olefination process. AB - [structure: see text] The synthesis of two truncated bryostatin analogues 2 and 3 is described. High-field NMR measurements on the C-ring analogue 3 in C(2)H(3)CN containing 25% (2)H(2)O have shown that it binds to the CRD2 of human PKC-alpha at virtually the same position as phorbol-13-acetate (PA) and bryostatin 1 (1). NMR titration studies have also revealed that 3 binds to the CRD2 with a potency similar in magnitude to PA but much less potently than 1. PMID- 12583755 TI - Novel synthesis and reactions of 5,7-dialkyl-4,6-dioxo-4,5,6,7-tetrahydro- isothiazolo[3,4,-d]pyrimidine-3-carbonitriles and 6-methyl-4-oxo-4H-1-aza-5-oxa-2 thiaindene-3-carbonitrile. AB - [reaction: see text] 5,7-Dialkyl-4,6-dioxo-4,5,6,7-tetrahydroisothiazolo[3,4 d]pyrimidine-3-carbonitriles 4, prepared from 6-amino-1,3-dialkyluracils 3 and 4,5-dichloro-5H-1,2,3-dithiazolium chloride (Appel's salt) 1, are utilized for the preparation of new derivatives of 4 bearing amino, alkylthio, amido, thioamido, tetrazolyl, and carboximidic acid ethyl ester groups at position 3. Similarly, the reactions of 6-methyl-4-oxo-4H-1-aza-5-oxa-2-thiaindene-3 carbonitrile 8, prepared from 4-amino-6-methyl-2-pyrone 6 and 1, with alkyl- and arylamines in DMF at 50 degrees C and reflux afforded different isothiazole derivatives 11 and 17, respectively. On the other hand, treatment of 8 with 1,3 diaminopropane in THF at room temperature, followed by chromatography on silica gel, gave 3-(2-oxopropyl)-6,7,8-trihydro-4H-1-thia-2,5,9 triazacyclopentacyclononene-4,10-dione 12 in 59% yield. PMID- 12583754 TI - New, abridged pathway to Masamune's "southern hemisphere" intermediate for the total synthesis of bryostatin 7. AB - [reaction: see text] The "Southern Hemisphere" intermediate 2, used by Masamune and co-workers for their asymmetric total synthesis of bryostatin 7 (1), has been synthesized from (E)-1,4-hexadiene (11) by a 24-step pathway that has a longest linear sequence of only 20 steps. This is the shortest synthesis of 2 so far recorded, and moreover, it is fully stereocontrolled. PMID- 12583756 TI - Rhodium carbenoid N-H insertion reactions of primary ureas: solution and solid phase synthesis of imidazolones. AB - [reaction: see text] The solution and solid-phase synthesis of imidazolones is reported. The key step for the preparation of these compounds is the N-H insertion reaction of primary ureas into highly reactive rhodium carbenoid intermediates. Typically, a soluble or support-bound alpha-diazo-beta-ketoester is treated with a rhodium carboxylate catalyst in the presence of a primary urea to give the corresponding N-H insertion product. Subsequent acid-catalyzed cyclodehydration of these insertion products affords the desired imidazolone products. PMID- 12583757 TI - Halichondrin B: synthesis of a C1-C14 model via desymmetrization of (+) conduritol E. AB - [reaction: see text] A model C1-C14 segment (1) of halichondrin B was synthesized from (+)-conduritol E (7) in 18 steps and 2.9% overall yield. Key features of the synthesis include the novel ozonolytic desymmetrization of C(2)-symmetric diol 6, the early-stage construction of the C-ring which accompanies installation of the crucial C12 stereocenter, and the use of an enol ether C14-ketone surrogate as a precursor to the CDE-"caged" ketal. PMID- 12583758 TI - Highly stereoselective aldol reactions of titanium enolates from lactate-derived chiral ketones. AB - [reaction: see text] Highly stereoselective titanium-mediated aldol reactions based on lactate-derived ketones are reported. The stereochemical outcome of the process depends on the protecting group (PMB or Bn) and the Lewis acid (i PrOTiCl(3) or TiCl(4)) used in the enolization step, the corresponding anti-syn or syn-syn aldols being prepared in high yields and with diastereomeric ratios up to 99:1. PMID- 12583759 TI - Efficient convergent synthesis of 1alpha,25-dihydroxyvitamin D3 and its analogues by Suzuki-Miyaura coupling. AB - [reaction: see text] 1alpha,25-Dihydroxyvitamin D(3) was synthesized by the Suzuki-Miyaura coupling of the A-ring intermediate 1, which was efficiently prepared from readily available 1,7-enyne 2, with the corresponding boronate compound of the C,D-ring portion. The method was applied to prepare des-C,D analogues of 1alpha,25-dihydroxyvitamin D(3). PMID- 12583760 TI - Stereoretentive C-H bond activation in the aqueous phase catalytic hydrogenation of amino acids to amino alcohols. AB - [reaction: see text] At 100 degrees C and 1000 psi of hydrogen, aqueous l-alanine undergoes facile hydrogenation to l-alaninol over a 5% Ru/C catalyst. In the presence of added acid to protonate the carboxylate moiety, the reaction is faster and more selective than analogous reductions of simple alkanoic acids. Stereochemistry at the alpha-carbon is retained despite complete exchange of hydrogen at this site, as shown by deuterium incorporation. Similar stereoretentive C-H bond activation at C2 is seen in l-alaninol itself, and when acid is omitted, in l-alanine. These processes reveal a class of mild, highly stereoretentive C-H bond activations occurring in water over a heterogeneous catalyst. PMID- 12583761 TI - An efficient method for removal of ruthenium byproducts from olefin metathesis reactions. AB - [reaction: see text] Sequential treatment of the ring-closing metathesis reaction products with silica gel, activated carbon (50 equiv wt relative to the crude products), and column chromatography on silica gel efficiently removed dark brown ruthenium byproducts from the reaction mixture. After this treatment, colorless compounds could be obtained with a ruthenium level of 0.06-0.53 microg per 5 mg of product. PMID- 12583762 TI - Asymmetric total synthesis of the proposed structure of the medicinal alkaloid jamtine using the chiral base approach. AB - [reaction: see text] A highly step-economic asymmetric synthesis of the tetracyclic structure assigned to the medicinal alkaloid jamtine has been accomplished using a chiral lithium amide base desymmetrization of a ring-fused imide. The structure synthesized appears to be different from that of the natural product originally reported. PMID- 12583763 TI - Synthesis of a carbohydrate-derived hydroxamic acid inhibitor of the bacterial enzyme (LpxC) involved in lipid A biosynthesis. AB - [reaction: see text] The enzyme LpxC (UDP-3-O-[(R)-3-hydroxymyristoyl]-GlcNAc deacetylase) catalyzes the second step of lipid A biosynthesis and is essential for bacterial growth. A GlcNAc-derived hydroxamic acid inhibitor 8 of this enzyme was synthesized using two different routes. Compound 8 exhibits activity toward LpxC enzymes from a wider spectrum of bacterial species than any of the previously reported hydroxamic acid inhibitors. PMID- 12583764 TI - Novel synthesis of 2-arylbenzothiazoles mediated by ceric ammonium nitrate (CAN): a rebuttal. AB - [reaction: see text] The reported synthesis of 2-arylbenzothiazoles mediated by ceric ammonium nitrate (CAN) is irreproducible; the only products formed in this reaction are bis-(p-tolyl) disulfide and p-tolyl p-toluenethiosulfonate. PMID- 12583765 TI - Novel sulfinyl imine ligands for asymmetric catalysis. AB - [structure: see text] A novel class of P,N-sulfinyl imine ligands has been prepared that incorporates chirality solely at sulfur. The Pd complex of ligand 14 catalyzes the allylic alkylation reaction with high enantioselectivity (94%), and the first crystal structure of a Pd-bound sulfinyl imine provides insight into binding mode and origins of stereoselectivity. PMID- 12583766 TI - En route to archimedene: total synthesis of C3h-symmetric [7]phenylene. AB - [reaction: see text] The total synthesis of C(3)(h)-symmetric [7]phenylene has been accomplished by triple cobalt-catalyzed cycloisomerization of an appropriate nonayne. Its spectral data are in accord with the expectations for a triply angularly fused system, but its calculated heat of formation suggests the presence of a destabilizing sigma effect relative to its D(3)(h) isomer. The molecule constitutes the largest substructure of archimedene hitherto synthesized. PMID- 12583767 TI - First direct reductive amination of mucochloric acid: a simple and efficient method for preparing highly functionalized alpha,beta-unsaturated gamma butyrolactams. AB - [reaction: see text] The first direct reductive amination of mucochloric acid (1) has been accomplished. Reaction of 1 with various alkyl, aryl, and benzylamines, followed by reduction in the same pot, provides an efficient method of obtaining N-benzyl-3,4-dichloro-1,5-dihydro-pyrrol-2-one and N-aryl (or alkyl)-3,4-dichloro 1,5-dihydro-pyrrol-2-ones. PMID- 12583768 TI - Addition reaction of azido-exTTFs to C60: synthesis of fullerotriazoline and azafulleroid electroactive dyads. AB - [structure: see text] The addition reaction of azido-exTTFs to C(60) affords electroactive fullerotriazoline and azafulleroid dyads, which behave as amphoteric redox systems. Fluorescence experiments and transient absorption spectroscopy reveal that excitation of the fullerene moiety leads to radical pair lifetimes that are 2 orders of magnitude higher than those previously reported for related fullerotriazolines. PMID- 12583769 TI - Catalysts with mixed ligands on immobilized supports. Electronic and steric advantages. AB - [reaction: see text] Immobilized dirhodium(II) catalysts having mixed chiral ligands enhance reactivity (AH = azetidinone) and influence stereoselectivity in cyclopropanation and carbon-hydrogen insertion reactions. PMID- 12583770 TI - Development of novel diastereoselective alkenylation of enolates using alkenylselenonium salts. AB - [reaction: see text] A novel alkenylation of enolates using alkenylselenonium salts is described. A reaction of lithium enolates, which were prepared in situ by the reaction of LiHMDS and carbonyl compounds, with alkenylselenonium salts gave the ethenylation products of carbonyl compounds in high yield. Diastereoselective alkenylation was also accomplished by the reaction of the enolates derived from N-acyl-1,3-oxazolidin-2-ones with the alkenylselenonium salt to afford good results (up to 92% yield and up to 95% de). PMID- 12583771 TI - Chemoselective radical cleavage of Cbz-protected nitrogen compounds. AB - [reaction: see text] Tributylstannyl radicals promote the deprotection of N-Cbz derivatives of amides and nitrogen-containing heteroaromatic rings. These radical conditions do not affect N-Cbz derivatives of basic amines. PMID- 12583772 TI - Stereoselective synthesis of (Z)-2-fluoro-1-alkenyl(phenyl)iodonium tetrafluoroborates. AB - [reaction: see text] (Z)-2-Fluoro-1-alkenyl(phenyl)iodonium salts were stereoselectively prepared by the reaction of alkynyl(phenyl)iodonium salts with aqueous HF in good yields. The method is applicable to the synthesis of fluoroalkenyliodonium salts having functional groups such as ketone, ester, and chloride. (Z)-2-Fluoro-1-alkene, (Z)-2-fluoro-2-alkenoate, and (Z)-beta fluoroenyne could be stereoselectively prepared from the fluoroalkenyliodonium salt. PMID- 12583773 TI - A mild, chemoselective protocol for the removal of thioketals and thioacetals mediated by Dess-Martin periodinane. AB - [reaction: see text] This paper describes the development of a useful procedure for the removal of thioacetals and thioketals using Dess-Martin periodinane (DMP) reagent. In contrast to existing methods, this protocol offers general reactivity, compatibility with a wide range of functional groups, and convenient reaction times. Also discussed are chemoselectivity experiments involving functionalities that may be subject to oxidation by DMP, qualitative effects of substrate on hydrolysis rate, and direct thioacetal to acetal conversions. PMID- 12583774 TI - The Lewis acids catalyzed aza-Diels-Alder reaction of methylenecyclopropanes with imines. AB - [reaction: see text] The methylenecyclopropanes can react with various imines to give the aza-Diels-Alder adducts in good yields in the presence of Lewis acids. PMID- 12583775 TI - Epoxide-initiated electrophilic cyclization of azides: a novel route for the stereoselective construction of azabicyclic ring systems and total synthesis of (+/-)-indolizidine 167B and 209D. AB - [reaction: see text] A novel and general method for the stereoselective construction of 5-hydroxymethyl azabicyclic ring skeletons based on epoxide initiated electrophilic cyclization of azides has been developed and applied in the synthesis of (+/-)-indolizidine 167B and 209D with an overall yield of 16.5% and 17.8%, respectively. The efficiency of this methodology is further exemplified in the synthesis of azepine skeleton via tandem cation-olefin-azide cyclization. PMID- 12583776 TI - Total synthesis of (-)-21-isopentenylpaxilline. AB - [structure: see text] The total synthesis of (-)-21-isopentenylpaxilline (1) has been achieved. Key elements of the synthesis include the stereocontrolled construction of the advanced eastern hemisphere (-)-5, a highly efficient union of the eastern and western fragments (-)-5 and 4, respectively, exploiting our 2 substituted indole synthesis, and a new protocol for the construction of ring C. PMID- 12583777 TI - Anti-selective aldol reactions with titanium enolates of N glycolyloxazolidinethiones. AB - [reaction: see text] A highly diastereoselective anti aldol addition utilizing a variety of N-glycolyloxazolidinethiones has been developed. Enolization of an N glycolyloxazolidinethione with titanium (IV) chloride and (-)-sparteine followed by addition of an aldehyde activated with additional TiCl(4) resulted in highly anti-selective aldol additions, typically with no observable syn isomers. Allyl protected glycolates demonstrated the highest levels of selection and yields, although O-benzyl and O-methyl glycolyloxazolidinethiones also performed well. PMID- 12583778 TI - Probing electronic effects in the asymmetric Heck reaction with the BIPI ligands. AB - [structure: see text] The new BIPI ligands are phosphinoimidazolines that can be electronically tuned in three different ligand regions to explore electronic effects in asymmetric catalysis. Their application to the asymmetric Heck reaction (AHR) in the creation of a chiral quaternary center is described. Enantioselectivity is shown for the first time to depend linearly on phosphine electron density. Changing the ligand basicity by variation of the R(2) or R(3) substituents reverses facial selectivity. PMID- 12583779 TI - Toward the synthesis of peloruside a: fragment synthesis and coupling studies. AB - [reaction: see text] The asymmetric synthesis of building blocks 3, 4, and 5, corresponding to C(12)-C(19), C(7)-C(11), and C(1)-C(6) segments of peloruside A, is reported, along with boron-mediated aldol coupling studies directed toward the assembly of the complete carbon skeleton of this microtubule-stabilizing macrolide. PMID- 12583781 TI - High-frequency ultrasonographic evaluation of conjunctival intraepithelial neoplasia and squamous cell carcinoma. AB - OBJECTIVE: To evaluate the high-frequency B-scan ultrasonographic characteristics of squamous conjunctival neoplasia (conjunctival intraepithelial neoplasia and squamous cell carcinoma). METHODS: Each of 11 patients was examined with 20- and/or 50-MHz ultrasonography in a retrospective consecutive case series. MAIN OUTCOME MEASURES: Ultrasonographic findings with clinical and histopathologic correlations. RESULTS: Eleven eyes of 11 patients (8 men) were affected. Disease involved the right eye in 6 (55%) of the patients and the left eye in 5 (45%) of the patients; it was multifocal in 5 (45%) of the eyes. All tumors extended to, or primarily involved, the limbal conjunctiva. One patient developed superficial spread overlying a functioning partial-thickness filtering bleb, 1 developed intraocular extension, 1 developed scleral invasion, and 3 developed orbital involvement before treatment. Results of ultrasonographic examinations showed that the superficial aspect of the smaller limbal tumors appeared as fusiform thickening of the conjunctiva. In all patients, the tumor surface was highly reflective in contrast to the characteristically low reflectivity seen within the tumor stroma. Intraocular tumor extension was variably reflective, but evidenced by blunting of the anterior chamber angle and thickening of the uvea. Orbital extension was viewed as low reflective tumor extension into the relatively hyperechoic orbital tissues. CONCLUSIONS: High-frequency ultrasonography may be used to assess the extent of squamous conjunctival neoplasia. While the 50-MHz system offered better resolution, 20-MHz ultrasonography allowed for a wider and deeper field of view. High-frequency ultrasonography was useful in determining tumor thickness, shape, and internal reflectivity, and especially in revealing tumor extension into the sclera, eye, and orbit. PMID- 12583782 TI - Differentiation of lipid tear deficiency dry eye by kinetic analysis of tear interference images. AB - OBJECTIVE: To use kinetic changes to characterize tear interference images in patients with lipid tear deficiency (LTD) dry eye. METHODS: We used a DR-1 camera to digitize and analyze sequential images of tear interference on the central 8 mm of the cornea at the start of complete blinking in 11 healthy volunteers and 8 patients with LTD and noninflamed meibomian gland dysfunction. MAIN OUTCOME MEASURES: We studied tear lipid spread time and pattern, stability of the lipid after spread, and distribution of thickness in a prospective, case-control study. RESULTS: On complete lid closure, the lipid spread was horizontal in healthy eyes but vertical in LTD (P<.001). Mean +/- SD lipid spread time was 0.36 +/- 0.22 seconds in healthy eyes but 3.54 +/- 1.86 seconds in LTD (P<.001). Conventional DR-1 grading could not distinguish these groups (P =.32). Mean +/- SD lipid film thickness in healthy eyes was 74.5 +/- 6.9 nm, thicker than the 43.8 +/- 10.6 nm in LTD (P<.001), and this result was confirmed by qualification with intensity histogram (P<.001). CONCLUSIONS: Kinetic analysis of the tear interference revealed distinctive differences in the time and pattern of lipid spread and the distribution and stability of resultant lipid thickness between healthy subjects and patients with LTD. This method can be coupled with others for formulating effective therapies for patients with dry eye. PMID- 12583783 TI - Vitreous amino acid concentrations in patients with glaucoma undergoing vitrectomy. AB - OBJECTIVE: To measure vitreous concentrations of glutamate and other amino acids in patients with glaucoma undergoing vitrectomy. METHODS: Undiluted vitreous samples were collected from patients undergoing vitrectomy at the University of Iowa (Iowa City) between 1997 and 1998 (n = 69). Vitreous concentrations of 16 amino acids, including glutamate, were determined using high-pressure liquid chromatography. Patients with a history of diabetes mellitus were excluded from the analysis. The study group consisted of those with a history of glaucoma (n = 8), and the control group included those with an epiretinal membrane and/or macular hole with no history of glaucoma (n = 17). Comparison of amino acid concentrations between the 2 groups was performed using a multifactor main effects model that adjusted for the effect of 10 selected covariates. Power analysis was done to determine the level of significant difference in amino acid concentrations. RESULTS: The glaucoma group comprised vitreal specimens from patients with primary open-angle (n = 3) and angle-closure glaucomas that included aqueous misdirection (n = 2), uveitis with secondary angle-closure (n = 2), and Axenfeld Rieger syndrome (n = 1). Indications for vitrectomy in this group included epiretinal membrane, retinal detachment, aqueous misdirection, and uveitis. The control group included specimens from patients with a macular hole (n = 11) and epiretinal membrane (n = 7), with 1 eye having both. Surgical indications in controls were macular hole, retinal detachment, and epiretinal membrane. The mean +/- SD levels of vitreous glutamate, glycine, gamma aminobutyric acid, and alanine were 6.1 +/- 2.4, 16.3 +/- 7.5, 0.8 +/- 0.3, and 260.5 +/- 101.9 microM, respectively, in glaucoma and 5.2 +/- 2.3, 8.5 +/- 2.5, 0.6 +/- 0.2, and 159.5 +/- 54.9 microM in controls (P >.05 for all). None of the 16 amino acid concentrations measured showed a statistically significant difference between glaucoma and controls (P values between.06 and >.99). A power analysis indicated that a 1.8-fold elevation in the glutamate level was needed to reach significance. MAIN OUTCOME MEASURES: Vitreous amino acid concentrations. CONCLUSIONS: None of the 16 amino acids measured, including glutamate, were significantly elevated in the vitreous of glaucomatous eyes compared with controls. Our results are not consistent with the simple hypothesis of glutamate excitotoxicity in glaucoma. Instead, our findings indicate the dynamic nature of extracellular glutamate, whose concentration is dependent on complex mechanisms not yet fully understood. Further studies are needed to fully elucidate the role of glutamate in the pathogenesis of glaucoma. PMID- 12583784 TI - Scanning laser polarimetry and detection of progression after optic disc hemorrhage in patients with glaucoma. AB - OBJECTIVE: To examine retinal nerve fiber layer changes with scanning laser polarimetry (SLP) in the eyes of patients with glaucoma and optic disc hemorrhage. METHODS: Automated perimetry and SLP were performed in 17 eyes of 17 patients identified prospectively with optic disc hemorrhage. Criteria for visual field progression were based on decreased sensitivity seen at 3 adjacent points on the total deviation plot. Progression on SLP images was defined as a 15% or more decrease in the average thickness of the affected quadrant (superior or inferior), a 25% or more reduction in the affected quadrant ratio, an increase in the nerve fiber analyzer number of 10 or more (GDx Nerve Fiber Analyzer; Laser Diagnostic Technologies), or any change on Serial Analysis of the SLP images. MAIN OUTCOME MEASURES: Visual field progression and SLP image progression. RESULTS: The mean follow-up was 31 months (minimum, 12 months). Of the 17 eyes, 10 (59%) had visual field progression. No significant change was seen on SLP images for either the total group or the group with visual field progression. Five eyes (29%) showed progression on SLP images, 3 of which also showed visual field progression. Ten eyes showed progression on SLP images that was not confirmed on subsequent imaging. CONCLUSIONS: In eyes with visual field progression after optic disc hemorrhage, a significant change in the SLP image was not seen. Fluctuation of SLP results in patients with glaucoma necessitates confirmation of progression seen on SLP images. PMID- 12583785 TI - Peripapillary detachment in pathologic myopia. AB - OBJECTIVE: To describe peripapillary detachment in pathologic myopia (PDPM), a newly recognized fundus lesion. DESIGN: Retrospective medical record review. METHODS: We evaluated a series of myopic eyes that had a yellow-orange elevation of the retina and retinal pigment epithelium at the inferior border of the myopic conus. RESULTS: Twenty eyes of 15 patients were identified during a 17-year period to have characteristic findings of PDPM. The mean age of the patients was 58 years. They were followed up for an average of 6 years. The mean spherical equivalent correction was -11.00 diopters (D) (range, -6.00 to -16.00 D). The mean axial length was 27.4 mm (range, 25.3-28.9 mm). In each case, ophthalmic coherence tomographic examination showed a localized detachment of the retinal pigment epithelium and retina corresponding to the PDPM lesion. During the follow up period, the lesion remained stable in all cases except for 1. No apparent negative effect on visual function was noted. CONCLUSIONS: Peripapillary detachment in pathologic myopia is an asymptomatic, yellow-orange peripapillary detachment of the retinal pigment epithelium and retina in pathologic myopia. Recognition of this lesion is important to distinguish it from other fundus pathologic conditions, such as tumors or choroidal neovascularization, which require further investigation and treatment. PMID- 12583786 TI - Vitritis and chorioretinitis in a patient with West Nile virus infection. AB - OBJECTIVE: To describe the visual outcome, fundus appearance, and fluorescein angiographic findings of a patient diagnosed as having acute West Nile virus infection. METHODS: We conducted comprehensive eye examinations, including visual acuity testing and slitlamp and fundus examinations, along with fluorescein angiography at the initial and follow-up visits. RESULTS: A 62-year-old white woman had a 2-week history of floaters in her left eye along with symptoms of fatigue, a left-sided frontal headache, and a low-grade fever. She was found to have anterior uveitis, vitritis, and nonnecrotizing chorioretinitis in her left eye with similar but milder findings in the right eye. The anterior uveitis responded to topical steroids. Because of continued fevers and fatigue, a serologic test for West Nile virus immunoglobulin M was performed, and the results were found to be positive for this disease. CONCLUSION: These unique eye findings, along with associated systemic signs and symptoms, can potentially indicate infection with the West Nile virus. PMID- 12583788 TI - Glaucoma detection using scanning laser polarimetry with variable corneal polarization compensation. AB - OBJECTIVE: To compare the ability of scanning laser polarimetry (SLP) to discriminate between healthy and glaucomatous eyes with manufacturer-assumed fixed and subject-specific variable corneal polarization magnitude (CPM) and corneal polarization axis (CPA) values. METHODS: An SLP was modified to enable the measurement of CPM and CPA values so that compensation for corneal birefringence could be corrected on a subject-specific variable basis. We examined 40 healthy eyes and 54 glaucomatous eyes with repeatable visual field damage (average +/- SD mean deviation, -6.5 +/- 4.9 dB) were examined by SLP using the manufacturer-assumed fixed corneal compensation (FCC-SLP) values and subject-specific variable corneal compensation (VCC-SLP) values. Areas under the receiver operating characteristic (ROC) curve for discriminating between healthy and glaucomatous eyes using FCC-SLP and VCC-SLP parameters were compared. RESULTS: The areas under the ROC curve increased with VCC-SLP compared with FCC SLP, particularly for all thickness parameters. The parameters with which the area under the ROC curve improved significantly from FCC-SLP to VCC-SLP included average thickness (ROC curve area, 0.62 vs 0.75), superior integral (0.66 vs 0.79), ellipse average (0.65 vs 0.80), inferior average (0.66 vs 0.80), and superior average (0.68 vs 0.83). CONCLUSION: Variable corneal compensation to correct for subject-specific CPM and CPA can improve the ability of SLP to discriminate between healthy and glaucomatous eyes. PMID- 12583787 TI - National Eye Institute Visual Function Questionnaire in the Age-Related Eye Disease Study (AREDS): AREDS Report No. 10. AB - OBJECTIVES: To describe the vision-targeted, health-related quality of life, measured with the National Eye Institute Visual Function Questionnaire (NEI-VFQ), in patients with age-related macular degeneration, cataract, or reduced visual acuity; to determine the relationship between the NEI-VFQ subscale scores and clinical measures of visual function; and to assess the internal consistency and reliability of the NEI-VFQ subscales. DESIGN: The 39-item NEI-VFQ was administered at the 5-year clinic visit to 4077 Age-Related Eye Disease Study participants. RESULTS: The subscales of the NEI-VFQ had moderate to high internal consistency (Cronbach's alpha = 0.58-0.91). The NEI-VFQ scores for participants with advanced age-related macular degeneration in 1 or both eyes, severe nuclear opacity, reduced visual acuity, or cataract surgery generally were lower than scores for disease-free participants (P<.001). CONCLUSION: These findings support the use of the NEI-VFQ as a measure of vision-targeted, health-related quality of life among patients with age-related macular degeneration, cataract, or reduced visual acuity. PMID- 12583789 TI - Observation of choroidal circulation using index of erythrocytic velocity. AB - OBJECTIVE: To describe a noninvasive method to visualize choroidal circulation by means of erythrocytic velocity. METHODS: Laser speckle flowgraphy (LSFG) and indocyanine green (ICG) angiography were performed on 9 volunteers. The LSFG measures the quantitative relative velocity index of erythrocytes (normalized blur [NB] value) in retinal and choroidal vessels. We averaged NB values from 3 pulsations and made composite 1.5-mm-square NB maps during 1 pulsation. By overlapping 5 adjacent maps, we created a panoramic 3.0-mm-square NB map of the posterior pole. The vascular patterns of the panoramic map and ICG angiography were compared. To determine the influence of retinal vessels, we induced branch retinal artery occlusion in 2 monkey eyes and compared the panoramic maps before and after occlusion. RESULTS: The NB map showed pulsatile blood flow in choroidal and retinal vessels. Vascular pattern contrast was improved in the NB map. Choroidal vessels in ICG angiography corresponded to those in the NB map. Vascular patterns in the map changed little before and after branch retinal artery occlusion. CONCLUSIONS: The LSFG noninvasively visualized the hemodynamics of choroidal circulation, and the vascular pattern, which is mainly choroidal in origin, was comparable with that of ICG angiography. CLINICAL RELEVANCE: The LSFG may be used to evaluate choroidal hemodynamics in various choroidal diseases. PMID- 12583790 TI - In vivo imaging of human retinal flow dynamics by color Doppler optical coherence tomography. AB - BACKGROUND: Color Doppler optical coherence tomography (CDOCT) combines laser Doppler velocimetry and optical coherence tomography for simultaneous micron scale resolution cross-sectional imaging of tissue microstructure and blood flow. Recently, CDOCT was adapted to a slitlamp biomicroscope for imaging structure and blood flow in the human retina. OBJECTIVE: To demonstrate feasibility of CDOCT for imaging retinal hemodynamics. DESIGN: Enabling CDOCT to measure retinal blood flow pulsatility in humans. SETTING: Laboratory. MAIN OUTCOME MEASURES: Time resolved flow profiles and images of retinal blood flow dynamics for measurement of pulsatility within retinal vessels. RESULTS: Rapid sequences of images were acquired over selected vessels near the optic nerve head. From these images, retinal blood flow profiles were extracted and synchronized to an external reference obtained with a photoplethysmograph. Each profile was acquired in less than 10 milliseconds. CONCLUSIONS: Our results indicate that CDOCT provides laser Doppler information in addition to conventional optical coherence tomography, allowing the observation of blood flow dynamics simultaneous to imaging retinal structure. CDOCT is a promising technology for research and clinical studies of retinal blood flow dynamics. CLINICAL RELEVANCE: Blood flow dynamics, such as pulsatility and autoregulation, have been shown to change throughout the progression of diabetic retinopathy and glaucoma. Enabling CDOCT to observe retinal dynamics improves its potential as a clinical diagnostic tool. PMID- 12583791 TI - Corneal thickness and intraocular pressure in the Barbados eye studies. AB - OBJECTIVES: To describe the distribution of central corneal thickness (CCT) and evaluate its relationship to intraocular pressure (IOP) in the predominantly black population of the Barbados Eye Studies (BES). METHODS: Participants received a standardized examination, including pachymetry, applanation tonometry, and a comprehensive ophthalmologic evaluation. Analyses were based on data from all eyes, and generalized estimating equation methods were applied to account for the correlation between eyes. RESULTS: Among the 1142 consecutive participants with pachymetry measurements, the mean age was 64.3 years, and 58% were women. Black participants tended to have thinner corneas (mean thickness, 529.8 microm) than mixed (black and white) (537.8 microm) and white participants (545.2 microm), respectively. Among black participants, increasing values of CCT were significantly related to younger age (P<.001), diabetes history (P =.03), and refractive error (P =.03); a marginally significant relationship (with thinner corneas) was found with a clinical diagnosis of glaucoma (P =.07). Intraocular pressure was not associated with CCT in this population. CONCLUSIONS: Although other studies have reported a positive correlation between CCT and IOP, such a relationship was not substantiated in the black BES population. Black participants tended to have thinner corneas than white participants, whereas younger individuals, as well as those with a history of diabetes and more positive refractive errors, had thicker corneas. PMID- 12583792 TI - Risk factors for incident retinopathy in a diabetic and nondiabetic population: the Hoorn study. AB - OBJECTIVE: To investigate the effect of glycosylated hemoglobin, age, sex, hypertension, body mass index, waist-hip ratio, serum lipid levels, and smoking on the incidence of retinopathy in persons with normal and abnormal glucose metabolism. METHODS: The incidence of retinopathy was determined in 233 individuals, aged 50 to 74 years, by ophthalmoscopy and fundus photography at baseline and after an average follow-up of 9.4 years. Relative risks for retinopathy, estimated by odds ratios, were calculated for tertiles of cardiovascular risk factors at baseline. Logistic regression analysis was used, without and with adjustment for age, sex, hypertension, and glucose metabolism. RESULTS: The cumulative incidences of retinopathy among individuals with normal, impaired, and diabetic glucose metabolism were 7.3%, 13.6%, and 17.5%, respectively. Adjusted odds ratios for retinopathy were 2.36 (95% confidence interval, 1.02-5.49) for hypertension and 3.29 (95% confidence interval, 1.11 9.72) and 8.67 (95% confidence interval, 1.85-40.60) for the highest tertiles of glycosylated hemoglobin level and waist-hip ratio, respectively. No consistent or statistically significant associations with retinopathy were present for age, sex, body mass index, smoking, and serum levels of triglycerides and total, high density lipoprotein, and non-high-density lipoprotein cholesterol (P>.05 for all). CONCLUSION: Glycemia, hypertension, and abdominal obesity are determinants for retinopathy in a general population. PMID- 12583793 TI - Gene expression in donor corneal endothelium. AB - OBJECTIVE: To report gene expression profiles of normal human corneal endothelium with microarray analysis and serial analysis of gene expression (SAGE). METHODS: Corneal endothelium was removed from normal human corneas obtained from eye banks. Total RNA was isolated and SAGE analysis was performed. The same RNA source was used to construct a complementary DNA library that was hybridized to microarrays containing 12 558 transcripts. RESULTS: A total of 9530 SAGE tags were sequenced, representing 4724 unique tags. Microarray analysis identified 542 distinct transcripts. A database of human corneal endothelial gene expression was compiled. Of the SAGE tags, 1720 matched known genes, 478 corresponded to expressed sequence tags, and 2526 had no known match to public databases. The 5 most abundantly expressed SAGE tags were cytochrome c oxidase subunit II, adenosine triphosphate synthase F(0) subunit 6, carbonic anhydrase XII, 12S ribosomal RNA, and ferritin, heavy polypeptide 1. Thirty-four percent of the transcripts (n = 1616) were specific to the corneal endothelium, when compared with other publicly available SAGE libraries. The 5 most abundant unique tags were keratin 12, angiopoietinlike factor, annexin A8, and 2 tags with no match to the database. Many endothelial pump function enzymes were confirmed, including several plasma membrane Na( +)/K(+) adenosine triphosphatases and a recently reported bicarbonate transporter. CONCLUSIONS: Corneal endothelial gene expression profiles by the current analysis provide an understanding of endothelial metabolism, structure, and function; enable comparisons to diseased endothelium; and provide baseline data that may lead to the discovery of novel endothelial genes. PMID- 12583794 TI - A new lamellar wound configuration for penetrating keratoplasty surgery. AB - A modified penetrating keratoplasty procedure with a new lamellar configuration of the surgical wound was performed on 8 eyes with endothelial decompensation. This technique allows complete suture removal by 3 months postoperatively, substantially shortening the time necessary for visual rehabilitation. Refractive astigmatism before and after suture removal was minimized to 4 diopters or less in all of the eyes in our preliminary series. Because the anterior surface of the donor button is smaller than the posterior one (diameter, 7.0 mm and 9.0 mm, respectively), more endothelial cells can be transplanted while maintaining the anterior graft surface at a safe distance from the corneoscleral limbus. Finally, no expensive instrumentation is required for this procedure except for an artificial anterior chamber if whole globes are not available. The surgical technique and clinical results are presented in this article. PMID- 12583795 TI - The strange report of Cheselden's iridotomy. PMID- 12583796 TI - Unusual superficial variant of granular corneal dystrophy with amyloid deposition. PMID- 12583797 TI - Endophthalmitis caused by Mycobacterium chelonae abscessus after intravitreal injection of triamcinolone. PMID- 12583798 TI - Clinicopathologic reports, case reports, and small case series: surgical removal and histopathologic findings of a subfoveal neovascular membrane associated with choroidal osteoma. PMID- 12583799 TI - Prostatic adenocarcinoma metastatic to the anterior uveal tract. PMID- 12583800 TI - Treatment of complex choristoma by excision and amniotic membrane transplantation. PMID- 12583801 TI - Aggressive conjunctival squamous cell carcinoma in a patient following liver transplantation. PMID- 12583802 TI - Angle-closure glaucoma associated with ciliary body detachment in patients using topiramate. PMID- 12583803 TI - Epithelial downgrowth following insertion of an ahmed glaucoma implant. PMID- 12583804 TI - Traumatic cataract after inadvertent laser discharge. PMID- 12583805 TI - Magnetic resonance imaging signs may antedate visual loss in chiasmal radiation injury. PMID- 12583806 TI - Recurrent visual loss in Leber hereditary optic neuropathy. PMID- 12583807 TI - Retained intraocular cilium in the presence of silicone oil. PMID- 12583808 TI - Computed tomographic scan of a dinosaur's skull: the optic canal. PMID- 12583809 TI - Antibiotic prophylaxis in clear corneal cataract surgery. PMID- 12583811 TI - Can you learn ethics? PMID- 12583812 TI - Poly(2-alkylacrylic acid) polymers deliver molecules to the cytosol by pH sensitive disruption of endosomal vesicles. AB - The permeability barrier posed by cell membranes represents a challenge for the delivery of hydrophilic molecules into cells. We previously proposed that poly(2 alkylacrylic acid)s are endocytosed by cells into acidified vesicles and are there triggered by low pH to disrupt membranes and release the contents of endosomes/lysosomes to the cytosol. If this hypothesis is correct, these polymers could be valuable in drug-delivery applications. The present paper reports functional comparisons of a family of three poly(2-alkylacrylic acid)s. Poly(2 propylacrylic acid) (PPAA), poly(2-ethylacrylic acid) (PEAA) and poly(2 methylacrylic acid) (PMAA) were compared in red-blood-cell haemolysis assays and in a lipoplex (liposome-DNA complex) assay. We also directly examined the ability of these polymers to disrupt endosomes and lysosomes in cultured human cells. Our results show that: (i) unlike membrane-disruptive peptides, the endosomal disruptive ability of poly(2-alkylacrylic acid)s cannot necessarily be predicted from their haemolytic activity at low pH, (ii) PPAA (but not PEAA or PMAA) potently facilitates gene transfection by cationic lipoplexes and (iii) endocytosed poly(2-alkylacrylic acid)s are triggered by luminal acidification to selectively disrupt endosomes (not lysosomes) and release their contents to the cytosol. These results will facilitate the rational design of future endosomal disrupting polymers for drug delivery. PMID- 12583813 TI - Differential interaction of the tyrosine phosphatases PTP-SL, STEP and HePTP with the mitogen-activated protein kinases ERK1/2 and p38alpha is determined by a kinase specificity sequence and influenced by reducing agents. AB - The protein tyrosine phosphatases (PTPs) PTP-SL, STEP and HePTP are mitogen activated protein kinase (MAPK) substrates and regulators that bind to MAPKs through a kinase-interaction motif (KIM) located in their non-catalytic regulatory domains. We have found that the binding of these PTPs to the MAPKs extracellular-signal-regulated kinase 1 and 2 (ERK1/2), and p38alpha is differentially determined by the KIM-adjacent C-terminal regions of the PTPs, which have been termed kinase-specificity sequences, and is influenced by reducing agents. Under control conditions, PTP-SL bound preferentially to ERK1/2, whereas STEP and HePTP bound preferentially to p38alpha. Under reducing conditions, the association of p38alpha with STEP or HePTP was impaired, whereas the association with PTP-SL was unaffected. On the other hand, the association of ERK1/2 with HePTP was increased under reducing conditions, whereas the association with STEP or PTP-SL was unaffected. In intact cells, PTP-SL and STEP distinctively regulated the kinase activity and the nuclear translocation of ERK1/2 and p38alpha. Our results suggest that intracellular redox conditions could modulate the activity and subcellular location of ERK1/2 and p38alpha by controlling their association with their regulatory PTPs. PMID- 12583814 TI - Improvements in spinner-flask designs for insect-cell suspension culture. AB - Experiments have been conducted to characterize the growth of Spodoptera frugiperda (fall armyworm) (Sf9) cells in standard and modified spinner designs. Results suggest that in standard spinner flasks growth is limited by mixing and oxygen transfer. Modifications to spinner geometry, baffling and agitator aspect ratio provided increases in growth rate and final cell density. Improvements in the performance of spinners, such as those described herein, have the potential to simplify a process and decrease the chance of contamination by decreasing the number of required manipulations and increase process productivity by decreasing inoculum train cycle times. PMID- 12583815 TI - Purification of soluble human epidermal growth factor (hEGF) from recombinant Escherichia coli culture broth by using expanded-bed adsorption chromatography. AB - Human epidermal growth factor (hEGF) secreted by recombinant Escherichia coli was purified from culture broth by expanded-bed adsorption (EBA) chromatography, strong anion-exchange chromatography and finally preparative reversed-phase HPLC (RP-HPLC). The EBA chromatography step simultaneously captured the hEGF by cationic exchanger and removed the cellular biomass from the diluted culture broth. This step was carried out at high throughput, and resulted in a high yield (>90%) and a purification factor of approx. 20-fold to >80% purity. Its process performance was well maintained during a 16-fold scale-up. After the successive purification steps of anion-exchange chromatography and RP-HPLC, the overall yield was approx. 84% and the purity was satisfactory (>99.5%). It was concluded that the purification process was very efficient and scaleable, warranting its implementation in large-scale manufacturing. PMID- 12583816 TI - The clinical effectiveness and cost-effectiveness of vinorelbine for breast cancer: a systematic review and economic evaluation. PMID- 12583817 TI - Clinical effectiveness and cost-effectiveness of growth hormone in adults in relation to impact on quality of life: a systematic review and economic evaluation. PMID- 12583818 TI - A systematic review update of the clinical effectiveness and cost-effectiveness of glycoprotein IIb/IIIa antagonists. PMID- 12583820 TI - Screening for hepatitis C among injecting drug users and in genitourinary medicine clinics: systematic reviews of effectiveness, modelling study and national survey of current practice. PMID- 12583819 TI - Clinical effectiveness and cost-consequences of selective serotonin reuptake inhibitors in the treatment of sex offenders. PMID- 12583822 TI - How important are comprehensive literature searches and the assessment of trial quality in systematic reviews? Empirical study. PMID- 12583821 TI - A comparative study of hypertonic saline, daily and alternate-day rhDNase in children with cystic fibrosis. PMID- 12583823 TI - Tumor necrosis factor-based isolated limb perfusion for soft tissue sarcoma and melanoma: ten years of successful antivascular therapy. PMID- 12583824 TI - Pegylated interferon alfa-2b as treatment of patients with solid tumors. PMID- 12583825 TI - Tubulin interacting agents: novel taxanes and epothilones. AB - The microtubule cytoskeleton plays an important part in cell mitosis, which relies on polymerization and depolymerization of the protein tubulin. Such established drugs as the vinca alkaloids and colchacine work by inhibiting microtubule assembly, as does paclitaxel. This article describes a variety of promising novel taxanes and epothilones with similar mechanisms of action that are in various states of preclinical and clinical development. PMID- 12583826 TI - Preclinical and clinical evaluation of farnesyltransferase inhibitors. AB - Farnesylation of Ras, a protooncogene that is frequently mutated in a number of malignancies, is critical for its biologic function. This observation has led to the development of several agents that inhibit farnesyltransferase, known as farnesyltransferase inhibitors (FTIs). The antiproliferative and antitumor effects of these agents have been demonstrated in preclinical and clinical studies. Interestingly, FTI activity does not necessarily rely on ras mutational status, indicating that Ras is not the only FTI target. Clinical data suggest that FTIs, alone and in combination with other agents, have antitumor activity. Further study is needed to determine the precise mechanism of FTI antitumor activity as well as how and where FTIs will be best used clinically. PMID- 12583827 TI - New and modified interferon alfas: preclinical and clinical data. AB - Recombinant human interferon (IFN)-alpha was the first biotherapeutic agent approved by the US Food and Drug Administration for the treatment of a human malignancy. Its efficacy has also been demonstrated for treatment of several viral diseases. The human genome codes for 12 IFN-alpha proteins, with IFN alpha 1 and IFN alpha-2 accounting for the majority of the naturally occurring IFN-as. However, only subspecies of IFN alpha-2, recombinant human IFN alpha-2a and IFN alpha-2b, are commercially available in the United States. Other species of IFN-a may demonstrate equivalent or improved efficacy and have more tolerable side effects. This article describes ongoing preclinical and clinical studies of several new and modified IFN-alphas. A current phase I trial of a human recombinant IFN alpha-1 is described. Basic pharmacokinetics and clinical studies of polyethylene glycol (PEG) IFN alpha-2b are reviewed as well. Lastly, two novel types of IFN-a, one gene shuffled and one hybridized with human albumin, are summarized. PMID- 12583830 TI - Intensity-modulated radiotherapy for carcinoma of the head and neck. AB - Intensity-modulated radiotherapy (IMRT) is the next evolutionary step of three dimensional conformal radiotherapy (3-DCRT). IMRT plans are generated with computer-optimized nonuniform radiation beam intensities incident on a target or patient. The dose distributions in IMRT show significantly higher conformality compared with other methods of external-beam radiation delivery, including conventional 3-DCRT. This significant conformality is advantageous in malignancies of the head and neck, where differential dosage is needed to deliver higher doses to the primary tumor and lower doses to areas of subclinical disease. Topics discussed in this review include target delineation, salivary gland sparing, and treatment of specific primary sites. Available data from the literature are presented, including patterns of failure after IMRT. To illustrate the potential role of IMRT in dose escalation and sparing of normal tissues, fractionation strategies in the definitive treatment of head and neck malignancies are presented. In addition, the role of IMRT in treatment of recurrent malignancies of the head and neck is explored. Finally, general concepts of helical tomotherapy are discussed. PMID- 12583828 TI - Novel antifolate drugs. AB - Antimetabolites are active chemotherapeutic agents for many solid tumor and hematologic malignancies. Folate antagonists, purine analogues, and pyrimidine analogues are the three main categories of antimetabolites. Methotrexate, the most studied folate antagonist, is effective in many malignancies. Methotrexate inhibits dihydrofolate reductase, which leads to accumulation of polyglutamated folates, causing further inhibition of thymidylate synthase and glycinamide ribonucleotide formyltransferase. Subsequently, the lack of reduced folate substrates impairs synthesis of purine nucleotides, thymidylate, and certain amino acids, which can lead to cell death. However, methotrexate resistance develops through several mechanisms, including decreased folate carrier-mediated membrane transport, dihydrofolate reductase gene amplification, specific transcription-translational modifications, and downregulation of intracellular methotrexate polyglutamation. Antifolate drug development has focused on agents designed to overcome different aspects of methotrexate resistance. This article reviews the enzymatic targets for antifolates, describes the known mechanisms of antifolate resistance, and summarizes the current development of novel antifolate agents. Discussed specifically are trimetrexate, edatrexate, raltitrexed, pemetrexed, ZD9331, lometrexol, LY309887, GW1843, OSI-7904(L), and nolatrexed, all of which have unique clinical pharmacology and are in various stages of development. The toxicity of antifolates has been sporadic and difficult to predict clinically. Supplementation with folic acid and vitamin B(12) has been shown to reduce the toxicity of pemetrexed without affecting efficacy and has increased the therapeutic index for this novel agent. PMID- 12583832 TI - The skull base, paranasal sinuses, and related malignancies. AB - Tumors of the anterior skull base most commonly originate from the nasal cavity and paranasal sinuses. Such early presenting signs and symptoms as nasal obstruction, epiphora, and epistaxis are frequently thought to have a benign pathology. Proptosis, vision changes, skin fixation, and cervical adenopathy are associated with advanced disease. Treatment of paranasal sinus tumors is complex, involving combined therapy and multidisciplinary teams. The particular method of treatment is determined by multiple factors, including tumor histopathology, age, health of the patient, and extent of disease. In the past several years, numerous approaches to the skull base have evolved, allowing otolaryngologists and head and neck surgeons to extend earlier operative indications and treat cancers that were previously deemed unresectable. This article reviews some of the current surgical and nonsurgical approaches to skull base tumors originating from the paranasal sinuses. PMID- 12583831 TI - The role of epidermal growth factor receptor in head and neck squamous cell carcinoma. AB - Growth factor receptors play a crucial role in the cell proliferation pathways involved in the development of cancer. One such receptor, the epidermal growth factor receptor (EGFR), is upregulated in many types of human tumors, particularly head and neck squamous cell carcinoma (HNSCC). EGFR overexpression in HNSCC has been the basis for investigation of therapeutic strategies that target EGFR. EGFR-blocking methods under evaluation involve immunotoxins, monoclonal antibodies, EGFR-specific tyrosine kinase inhibitors, and antisense approaches. These molecular targeting tactics have produced a number of agents that are currently in various stages of preclinical investigation, along with clinical trials to assess their potential as anticancer treatments. PMID- 12583833 TI - Chemoprevention of head and neck cancer. AB - Squamous cell carcinoma of the head and neck is the most common epithelial neoplasm of the upper aerodigestive tract and represents a major health concern in the United States and worldwide. Invasive squamous cell carcinoma is the end result of a multiyear, multistep process of accumulation of genetic and phenotypic damage. Chemoprevention is defined as the use of pharmacologic or natural agents that inhibit the development of invasive cancer whether by blocking the DNA damage that initiates carcinogenesis or by arresting or reversing the progression of premalignant cells in which such damage has already occurred. Chemoprevention is widely recognized as an important area of research in head and neck cancer. This article reviews the field of chemoprevention and recent advances in molecular epidemiology and genetics. Current clinical trials are described. PMID- 12583834 TI - Nutrition and head and neck cancer. AB - The role of nutrition in patients with head and neck cancer, specifically squamous cell carcinoma, is underappreciated. The composition of the diet can contribute to carcinogenesis, and specific nutrients may offer some protection in the presence of known carcinogens (ie, tobacco). Patients with head and neck cancer are frequently malnourished, which may have prognostic implications for the morbidity and outcome of therapies. Although the benefits of preoperative nutritional supplementation have been demonstrated only in severely malnourished patients, the use of immune-enhancing formulas may prove to be beneficial. Special consideration should be given to the nutritional needs and possible interactions of diet and therapy in patients receiving radiation and chemotherapy. Physicians should be cognizant of the widespread use of alternative diets and nutritional supplements that can be harmful and may interact with standard treatments. New knowledge regarding the role of nutrition in cancer offers hope for the nutritional chemoprevention of head and neck cancers. PMID- 12583835 TI - Stereotactic radiosurgery for patients with cancer of the head and neck. AB - Patients with recurrent cancer of the head and neck involving the base of the skull have limited treatment options. Stereotactic radiosurgery is currently being investigated as a viable modality of treatment for these patients. This paper reviews the basic principles and treatment procedures involved in stereotactic radiosurgery, as well as published reports on its use with these patients. Finally, questions to be addressed by future studies for clarification of the role of this technology are outlined. PMID- 12583836 TI - Fibromyalgia, fatigue, and headache disorders. AB - Fibromyalgia, chronic fatigue, and primary headaches are common and debilitating disorders, and their related symptoms of widespread pain, fatigue, and headache have complex interactions and different implications for classification, diagnosis, mechanisms, and treatment. The "continuum" or "spectrum" idea and the modular headache theory are fundamental concepts in understanding these interactions. The overlap between symptom-based conditions leads the reasons to consider them as "functional somatic syndromes." Management of these patients includes a correct diagnosis, appropriate investigation for associated conditions, adequate treatment, and considering the therapeutic opportunities and limitations the comorbid disorders may impose. PMID- 12583837 TI - Epidemiology of migraine and other types of headache in Asia. AB - Earlier studies have raised the issue that Asians have a much lower migraine prevalence than Westerners. This article reviews the recent epidemiologic studies of headache in Asia using International Headache Society (IHS) classification criteria. Except for the Korean study and the first Hong Kong study (1992 to 1993), the prevalence of migraine has been quite consistent, ranging from 8.4% to 12.7%. The sex-specific migraine prevalence was 11.3% to 14.4% in women and 3.6% to 6.7% in men. The prevalence of IHS tension-type headache has also been similar among these studies (15.6% to 25.7%). The consistency of the prevalence of migraine and tension-type headache among these Asian countries is interesting in a region where the cultural background and development are so diverse. These IHS migraine surveys show that migraine is a significant disease in Asia and that its prevalence rate is close to but in the low range of the those reported in Western countries. Two surveys of chronic daily headache conducted in Taiwan, one in the general population and the other in the elderly population, have also revealed prevalence rates (3.2% to 3.9%) similar to those of Western countries (3.0% to 4.7%). The geographic or ethnic diversity of migraine or other types of headache could be downplayed if a similar epidemiologic methodology and an identical case definition were used in the surveys among different regions of the world. PMID- 12583839 TI - Gene expression profiling to analyze embryonal tumors of the central nervous system. PMID- 12583838 TI - Pathophysiology of headache. AB - Clinical features of different headache syndromes have been described in great detail; however, its pathophysiology remains poorly understood. We review the latest findings in both human imaging studies and experimental animals to explain the possible mechanisms involved in the genesis of headache syndromes. PMID- 12583840 TI - Neurologic aspects of HIV infection in infants and children: therapeutic approaches and outcome. AB - Central nervous system (CNS) HIV-related disorders frequently have devastating consequences. Significant progress has been made in the early diagnosis and treatment of the HIV-infected patient. As a result, the prevalence and natural history of neurologic illnesses have changed. This paper reviews the epidemiology, clinical manifestations, and neuropathogenesis of HIV-related CNS disorders. Advances in antiretroviral therapy, neuroprophylaxis, and neuroprotective strategies are also discussed. PMID- 12583841 TI - Neurocognitive dysfunction in children with neurofibromatosis type 1. AB - The cognitive dysfunction associated with neurofibromatosis type 1 (NF1) is an intriguing aspect of this phenotypically heterogeneous genetic neurocutaneous disorder. A broad range of both nonverbal and verbal learning disabilities are evident in approximately 30% to 65% of children with NF1. Deficits in IQ, executive function, attention, and motor skills have also been documented. Current challenges lie in discovering the underlying multifactorial etiologies of the cognitive abnormalities found in NF1. Likely answers lie in neuroanatomic correlates as seen on neuroimaging as well as in molecular and genetic advances into the role of neurofibromin, the protein product of the NF1 gene. The development of NF1 animal models with learning and memory difficulties similar to those seen in humans demonstrates promising preliminary evidence that medical treatment of cognitive abnormalities may one day be possible. PMID- 12583842 TI - Neurologic complications of chemotherapy for children with cancer. AB - Survival rates for children with cancer have increased dramatically over the past few decades. Development of new chemotherapeutic agents and the expanded use of older agents have had a major impact on this celebrated improvement. Chemotherapy can have, however, significant toxicity on the nervous system. The most common neurologic complications involve acute alterations in consciousness, leukoencephalopathy, seizures, cerebral infarctions, paralysis, neuropathy, and ototoxicity. Most of the information on toxicity comes from prospective reports and the adult patient population. Methotrexate, cyclosporin, and platinum compounds are the most frequently cited. No prospective studies have been done to evaluate chemotherapy-induced neurotoxicity in the pediatric population, and the exact incidence of such complications is unknown. Such investigation is greatly needed, as it may lead to a better understanding of how chemotherapy affects the nervous system and ultimately help develop more strategies to prevent drug related neurotoxicity in pediatric cancer patients. PMID- 12583843 TI - Treatment approaches for children with Tourette's syndrome. AB - Tourette's syndrome has been of neurologic and psychiatric interest since the original description of this condition by Gilles de la Tourette in 1885, and it has been associated with a wide variety of treatments over the years. With the advent of advances in neurochemistry and neuropharmacology, a neurobiologic approach has emerged with the application of many drugs from the fields of neurology and psychiatry. In addition, many of the comorbid conditions that coexist, such as attention deficit disorder and obsessive compulsive disorder (OCD), are amenable to both pharmacologic and behavioral approaches. Drug treatment has included dopamine receptor blockers for tics, dopamine agonists, dopamine depletors, and stimulants for attention deficit hyperactivity disorder (ADHD), noradrenergic drugs for tics and ADHD, serotonergic drugs for OCD, and chemical denervation for involuntary movements with the use of botulinum toxin and stereotactic surgery. It is the purpose of this review to outline the various approaches that are currently available as treatments, realizing that as new drugs are introduced in neurology and psychiatry, they too will find their way into treatment options. PMID- 12583844 TI - The neurobiology of autism: new pieces of the puzzle. AB - The neurobiologic basis of autism is reviewed, with discussion of evidence from genetic, magnetic resonance imaging, neuropathology, and functional neuroimaging studies. Although autism is a behaviorally valid syndrome, it is remarkably heterogeneous and involves multiple developmental domains as well as a wide range of cognitive, language, and socioemotional functioning. Although multiple etiologies are implicated, recent advances have identified common themes in pathophysiology. Genetic factors play a primary role, based on evidence from family studies, identification of putative genes using genome-wide linkage analyses, and comorbidities with known genetic mutations. The RELN gene, which codes for an extracellular protein guiding neuronal migration, has been implicated in autism. Numerous neuropathologic changes have been described, including macroencephaly, acceleration and then deceleration in brain growth, increased neuronal packing and decreased cell size in the limbic system, and decreased Purkinje cell number in the cerebellum. Abnormalities in organization of the cortical minicolumn, representing the fundamental subunit of vertical cortical organization, may underlie the pathology of autism and result in altered thalamocortical connections, cortical disinhibition, and dysfunction of the arousal-modulating system of the brain. The role of acquired factors is speculative, with insufficient evidence to link the measles-mumps-rubella (MMR) vaccine with autism or to change immunization practices. PMID- 12583846 TI - Excessive daytime somnolence and increased rapid eye movement pressure in myotonic dystrophy. PMID- 12583845 TI - Organization of language networks in children: functional magnetic resonance imaging studies. AB - Functional magnetic resonance imaging (fMRI) is a relatively new neuroimaging procedure that has been used to study a wide variety of cognitive phenomena in adults, including attention, language, and memory. More recently, this technique has been successfully applied to pediatric populations as well. In particular, many investigators have employed fMRI as a tool to study language development in normal children. This paper reviews the current imaging research on the identification of cortex subserving components of language processing in young children. The literature suggests that fMRI can successfully identify regions of language cortex in children in much the same capacity as it can with adults, and that generally, adults and children show fundamental similarities in the patterns of activation. However, special considerations with pediatric imaging, paradigm design, and image analysis are also discussed. PMID- 12583847 TI - Sleep-related epilepsy. AB - Sleep is important for the general health of all, but is particularly essential for individuals with epilepsy. In these patients, a complex relationship exists between seizures and sleep, and both must be considered for optimal care. In this review, recent advances in the field of epilepsy and sleep are considered. These include sleep disorders with a specific relationship to epilepsy, the influences of sleep and sleep deprivation on interictal epileptiform discharges, detrimental effects of seizures on sleep, and the effects (good and bad) of epilepsy treatments on sleep. PMID- 12583850 TI - Reflections on the evolution of cardiology and cardiac surgery over the past 50 years. PMID- 12583848 TI - Sleep disturbances in Parkinson's disease. AB - Disorders of sleep and daytime alertness are frequent in Parkinson's disease patients and arise from a number of diverse factors. The most common complaint of night-time sleep disturbance in Parkinson's disease is sleep fragmentation. Sleep fragmentation can be associated with recurrent parkinsonian symptoms, the effect of medications, concomitant medical disorders such as nocturia, or psychiatric disorders such as depression or anxiety. Likewise, nocturnal sleep disturbance may arise from sleep apnea, periodic limb movements of sleep, or rapid eye movement (REM) sleep behavior disorder. Nocturnal sleep deprivation may lead to excessive daytime sleepiness. Other potential sources of daytime sleepiness include the effects of medications or disruption of central sleep mechanisms due to the pathologic processes of Parkinson's disease itself. Diagnosis of sleep disturbances and daytime sleepiness requires a direct interview of the patient and the caregiver, and may involve consultation with the sleep specialist or medical physician. Treatment is aimed toward improving night-time sleep and daytime drowsiness by addressing the causative factors. PMID- 12583849 TI - Hypnotics: an update. AB - This update reviews recent developments and advances in the therapeutic and side effect profile of the benzodiazepine receptor agonists (BZRAs), the generally accepted drug class of choice for the symptomatic treatment of insomnia. All the approved BZRAs, depending on their pharmacokinetic profile, improve and maintain sleep. The major recent advance is in the enhanced diversity of the pharmacokinetic profiles of these drugs, and thus in the flexibility available to the clinician in treatment strategy. Also, during the past decade the nature and significance of the side effects associated with the BZRAs and their determinants, dose and half-life, have been identified and clarified. The important remaining question is whether, and how, the efficacy and safety of the BZRAs change with chronic use. PMID- 12583851 TI - Calcific aortic stenosis: new pathophysiologic insights and possible new medical therapy. AB - Aortic stenosis is a progressive disease of aging with serious complications. A common disease of the elderly, it may inexorably progress to stenosis. Recent retrospective studies have correlated risk factors commonly associated with coronary and vascular atherosclerosis with an accelerated rate of aortic valve stenosis. Although hydroxymethyl glutaryl co-enzyme A reductase inhibitor (statin) treatment therapy has been shown to delay the rate of progression of valvular aortic stenosis, the salutary mechanism of the statin may be cholesterol lowering and/or anti-inflammatory. Further prospective studies are warranted to investigate the mechanism and medical therapy of aortic sclerosis and stenosis. PMID- 12583852 TI - Indications for surgery for aortic regurgitation. AB - Aortic valve replacement for isolated aortic regurgitation (AR) is usually not indicated unless the regurgitation is severe. However, not all patients with severe AR require aortic valve replacement. This review focuses on the causes of AR and the pathophysiology of acute versus chronic AR, and the attendant adaptive mechanisms of the left ventricle that ultimately determine their different natural histories. Aortic valve surgery must be performed in a timely manner to prevent cardiac death, ameliorate symptoms, and limit late postoperative excess mortality. PMID- 12583853 TI - Vena contracta width measurement: theoretic basis and usefulness in the assessment of valvular regurgitation severity. AB - In patients with valvular regurgitation, the regurgitation jet can be observed by Doppler color flow imaging. Vena contracta is defined as the narrowest part of the jet, just distal to the regurgitant orifice. Vena contracta dimensions reflect the severity of regurgitation. Vena contracta diameter, usually easy to measure in clinical practice, is well correlated with the effective regurgitant orifice area and the regurgitant volume. Cutoff values have been determined to identify severe regurgitation for mitral, aortic, and tricuspid valves. In clinical practice, determination of vena contracta diameter is a useful and simple method for assessment of valvular regurgitation. In the future, assessment of complex jet regurgitations will probably benefit from the contribution of three-dimensional Doppler flow imaging, which should improve the performances of the method. PMID- 12583854 TI - Trends in the surgical management of ischemic mitral regurgitation. AB - The surgical approach to ischemic mitral regurgitation (IMR) remains a topic of considerable controversy. Will coronary artery bypass alone suffice, or should the valve be intervened upon? The poor late survival of patients with IMR is well recognized, but it remains unknown if this can be altered by addressing the valve directly. And if surgery is undertaken, should the valve be repaired or replaced? The underlying mechanisms of IMR remain incompletely understood, and although current theory focuses on the role of alterations in ventricular geometry in its pathogenesis, IMR is most often addressed by annuloplasty alone. Is this sufficient, or does the ventricle itself require "remodeling?" The debate is confounded by imprecise terminology that fails to distinguish between acute and chronic disease, and active ischemia from completed infarction. Available clinical information is from retrospective studies with all of their inherent limitations and potential for bias. Still, progress is being made as increasing attention is focused on this clinically important entity. PMID- 12583855 TI - Is tissue-engineered heart valve replacement clinically applicable? AB - Tissue engineering of a heart valve has progressed dramatically in three different arenas: a biodegradable stented valve seeded with autogenous cells, decellularized allograft and xenograft valves that are seeded with autogenous cells, and decellularized allograft and xenograft valves that repopulate by adaptive remodeling in vivo. Preclinical evaluation and implants in sheep have been accomplished in each of these arenas, and clinical use of this emerging technology is occurring in the latter two. The clinical use of decellularized allografts that repopulate in vivo is expanding; however, its impact on allograft durability remains unknown. PMID- 12583856 TI - Embolic risk in subacute bacterial endocarditis: determinants and role of transesophageal echocardiography. AB - Embolic events are a frequent and life-threatening complication of subacute bacterial endocarditis (SBE), and are associated with a high mortality and morbidity. The prediction of the embolic risk in an individual patient remains a challenge. Echocardiography plays a key role in the management of infective endocarditis both for the diagnostic and prognostic assessment of these patients, but its value in predicting embolic events is still debated. This review focuses on the characteristics of embolic events in SBE, and on the role of echocardiography, especially transesophageal echocardiography (TEE), in the prediction of embolic events. The review confirms that, among other factors, TEE may be helpful in both the prediction of the embolic risk, and the decision to perform early surgery. PMID- 12583858 TI - Randomised trial of a perindopril-based blood-pressure-lowering regimen among 6105 individuals with previous stroke or transient ischaemic attack. PMID- 12583857 TI - Tinzaparin in acute ischaemic stroke (TAIST): a randomised aspirin-controlled trial. PMID- 12583859 TI - Ultrasound and angiography in the selection of patients for carotid endarterectomy. AB - The risk of ischemic stroke increases proportionately to the severity of carotid stenosis, and carotid endarterectomy is a durable procedure that reduces this risk. Although a combination of noninvasive tests, such as ultrasound and magnetic resonance angiography (MRA), have low misclassification rates compared with invasive angiography, the need for invasive angiography may not yet be obviated. Ultrasound appears to be a cost-effective screening strategy for a significant carotid stenosis that warrants angiographic confirmation and possible intervention. A combination of ultrasound and MRA appears to be the most common clinical pathway that can be accurate and cost-effective, if rigorous local validation of diagnostic criteria is performed. Ultrasound further supplements angiography by providing information about plaque morphology and physiologic measurements of collateralization of flow and vasomotor reactivity when additional tests, such as transcranial Doppler, are performed. Ultrasound and various angiographic imaging modalities have complementary value in patient selection for carotid endarterectomy. Currently, more invasive angiograms are being performed, due to a variety of new experimental interventions such as angioplasty and stenting, a subject of current clinical trials. PMID- 12583860 TI - The selection of antithrombotic agents in the prevention of recurrent ischemic stroke. AB - The scientific selection of antithrombotic therapy has been dominated by group based interpretation of data in the form of probability-based statistics in evidence-based medicine. Because the data in large randomized trials are grouped and averaged, the relationship to initial conditions of the patient is lost. There is a pathologic model and basis by which antithrombotic agents may be chosen for prevention of recurrent thrombus and thromboembolism in patients with stroke. This model applies in all settings, but has not been tested when the elements of the model remain connected to the individual patient and his or her unique context. Alternative math models and perception-based science respect the criticality of initial conditions and capture the rules that apply to the actual causal mechanisms within the patient's body. Individualized patient rather than group-based choices insure thrombus-type specific targeted therapy. PMID- 12583861 TI - Management of carotid artery stenosis: comparing endarterectomy and stenting. AB - Stroke ranks as the third leading cause of death, behind diseases of the heart and cancer. It is also the most important cause of disability. Approximately 750,000 people experience a stroke annually, costing an estimated $40 billion in direct and indirect costs. Approximately 25% of these ischemic events are related to occlusive disease of the cervical internal carotid artery. Carotid atherovascular stenosis increases the risk of ischemic stroke by acting as an embolic source, and causing hypoperfusion of the ipsilateral cerebral hemisphere. With some limitations, the North American Symptomatic Carotid Endarterectomy Trial (NASCET), European Carotid Surgery Trialists' Collaborative Group (ECST), and Asymptomatic Carotid Atherosclerosis Study (ACAS) have shown that carotid endarterectomy (CEA) substantially reduces the risk of stroke associated with certain grades of carotid stenosis. During the past few years, carotid angioplasty and stenting (CAS) has evolved as an alternative to CEA, particularly in patients who are known to have a higher complication rate with CEA. PMID- 12583862 TI - Mechanisms of ischemic brain damage. AB - Neurologic complications from cerebral ischemia occur frequently following cardiac arrest, as well as in the perioperative period in cardiac surgery. The cellular and molecular mechanisms of cerebral ischemia are complex. This article discusses several important cell death and salvage pathways that are important in experimental cerebral ischemia that may be critical to outcome in clinical brain injury. PMID- 12583863 TI - Congenital insensitivity to pain: an update. PMID- 12583864 TI - Glutamate-evoked pain and mechanical allodynia in the human masseter muscle. AB - The present study examined the effect of peripheral administration of the excitatory amino acid (EAA) glutamate on the intensity of perceived pain and pressure pain thresholds (PPTs) in healthy young women (n=17) and men (n=18). Two injections separated by 25 min of 0.2 ml, 1.0M glutamate into the masseter muscle produced significantly higher scores of pain on 0-10 cm visual analogue scales (VAS) in women than in men (analysis of variance, ANOVA: P<0.001). There was no significant difference between the VAS scores for the first and the second injections in either men or women. The PPTs determined in the masseter muscle were significantly reduced following the first injection and further significantly reduced after the second injection (ANOVA: P<0.001). Furthermore, the PPTs were reduced to a similar extent in both women and men (maximum 44-56%), suggesting that gender did not influence the process of sensitization. There were no significant difference in VAS scores or PPTs between women taking oral contraceptives (n=9) and those who did not (n=8) (ANOVAs: P=0.709, P=0.153). It is concluded that the VAS scores produced by intramuscular administration of 1.0M glutamate may reflect a gender-dependent activation of nociceptive pathways which, in part, may be mediated through peripheral EAA receptors. The reduction of PPTs in the masseter muscle following administration of glutamate in a concentration of 1.0M may reflect allodynia to mechanical stimuli. This process of sensitization was not gender-dependent. The present results suggest that injection of 1.0M glutamate into the masseter muscle may provide a useful experimental method to test sensitization and efficacy of peripheral EAA receptor antagonists in human subjects. PMID- 12583865 TI - Cold allodynia and hyperalgesia in neuropathic pain: the effect of N-methyl-D aspartate (NMDA) receptor antagonist ketamine--a double-blind, cross-over comparison with alfentanil and placebo. AB - Cold allodynia and hyperalgesia are frequent clinical findings in patients with neuropathic pain. While there have been several clinical studies showing the involvement of central sensitization mechanisms and N-methyl-D-aspartate (NMDA) receptor activation in mechanical allodynia/hyperalgesia and ongoing pain, the mechanisms of thermal allodynia and hyperalgesia have received less attention. The aim of the present study was to examine the effect of the NMDA-receptor antagonist ketamine on thermal allodynia/hyperalgesia, ongoing pain and mechanical allodynia/hyperalgesia in patients with neuropathic pain (11 patients with post-traumatic neuralgia and one patient with post-herpetic neuralgia). All the patients were known to suffer from severe cold allodynia (cold pain detection threshold (CPDT): 23.8 degrees C, median value). The mu-opioid agonist alfentanil was used as an active control. The study design was double-blind and placebo controlled and the drugs were administered i.v. (bolus dose and infusion). CPDT in the asymptomatic contralateral area was found to be significantly decreased (cold allodynia) compared to CPDT in site- and age-matched normal controls. Heat pain detection thresholds were found to be normal and no consistent heat hyperalgesia occurred. Alfentanil significantly reduced cold allodynia (by increasing CPDT) in symptomatic area (P=0.0076). Ketamine did not significantly increase the threshold. Significant and marked reductions of hyperalgesia to cold (visual analogue score at threshold value) were seen following both alfentanil (4.5 before, 1.4 after, median value) and ketamine (6.8 before, 0.4 after, median value). Alfentanil and ketamine also significantly reduced ongoing pain and mechanical hyperalgesia. It is concluded that NMDA-receptor mediated central sensitization is involved in cold hyperalgesia, but since CPDT remained unaltered, it is likely that other mechanisms are present. PMID- 12583866 TI - A dissociative change in the efficacy of supraspinal versus spinal morphine in the neuropathic rat. AB - The efficacy of spinally versus supraspinally administered morphine was studied in rats with a spinal nerve ligation-induced neuropathy. Behavioural assessment indicated that the effect of intrathecally administered morphine on pain-related responses was attenuated when compared with unoperated controls. The decreased efficacy of spinal morphine was associated with neuropathic symptoms, since sham ligation or nerve ligation without accompanying tactile allodynia did not lead to spinal inefficacy of morphine. In contrast, the pain attenuating effect of morphine in the periaqueductal gray (PAG) was enhanced in neuropathic animals. The effect of systemically administered morphine on pain-related behavior of neuropathic rats was in the same range as in controls or decreased, depending on the test. Coadministration of lidocaine or MK-801, a N-methyl-D-aspartate (NMDA) receptor antagonist, into the rostroventromedial medulla enhanced the tactile antiallodynic but not the thermal antinociceptive effect of intrathecally administered morphine in neuropathic animals. Supraspinal administration of MK 801 or lidocaine did not influence efficacy of spinal morphine in sham-operated animals. Electrophysiological recordings of nociceptive wide-dynamic range (WDR) neurons in the deep spinal dorsal horn of pentobarbitone-anesthetized animals corresponded to a large extent with behavioral results. The inhibitory effect of spinally and systemically administered morphine on WDR neuron responses was attenuated whereas that induced by morphine in the PAG was enhanced in neuropathic animals. The results indicate that in spinal nerve ligation-induced neuropathy the efficacy of spinal morphine is decreased whereas that of supraspinal morphine is increased. Descending influence from brainstem-spinal pathways, involving NMDA receptors in the rostroventromedial medulla, may contribute to the selective reduction in tactile antiallodynic efficacy of spinal morphine. PMID- 12583867 TI - Reflexes in sympathetic vasoconstrictor neurones arising from urinary bladder afferents are not amplified early after inflammation in the anaesthetised cat. AB - Pathophysiological processes in the viscera can lead to pain and hyperalgesia and exaggerated motility-regulating reflexes. This may be due to sensitisation of visceral afferents (peripheral sensitisation), which has repeatedly been shown to occur as a consequence of e.g. inflammation, and/or to sensitisation of dorsal horn neurones (central sensitisation), which is less well documented in the visceral domain. As an indicator of peripheral sensitisation, we previously analysed the responses of sacral spinal afferents after inflammation of the urinary bladder. Here, we studied reflexes in sympathetic vasoconstrictor neurones supplying skeletal muscle and skin elicited by bladder distension stimuli (vesico-sympathetic reflexes) before and after induction of bladder inflammation. Our aim was to test whether these vesico-sympathetic reflexes are amplified after inflammation in a way that would support a major functional role for post-inflammatory central sensitisation processes. Bladder inflammation was induced in anaesthetised cats by instillation of turpentine or mustard oil and vesico-sympathetic reflexes were studied 1 and 2 h after induction of the inflammation. Inflammation enhanced on-going activity in vasoconstrictor neurones supplying skeletal muscle (after 1 h to 187.6+/-36.8%, mean+/-SEM, P<0.01, and after 2 h to 139.1+/-12.9%, P<0.05, of baseline activity) and decreased it in most sympathetic neurones supplying skin (to 91.7+/-12.5%, P>0.05, and to 71.6+/ 11.3%, P<0.05, respectively, of baseline activity). Relative to the altered baseline activity vesico-sympathetic reflexes to graded distension of the inflamed bladder were quantitatively unchanged with a tendency to be diminished. Thus, the changes in on-going sympathetic vasoconstrictor activity and the distension-evoked reflexes directly mirrored the afferent input from the inflamed urinary bladder into the spinal cord, i.e. no increase of the gain of these reflexes was observed. These results suggest that in the first 2 h of inflammation, peripheral sensitisation processes play the main role for hyperalgesia and hyperreflexia of the urinary bladder. In contrast, central sensitisation appears to be of little importance during this time period. PMID- 12583868 TI - Gender differences in pressure pain threshold in healthy humans. AB - AIMS OF INVESTIGATION: To quantify the magnitude of putative gender differences in experimental pressure pain threshold (PPT), and to establish the relevance of repeated measurements to any such differences. METHODS: Two separate studies were undertaken. A pressure algometer was used in both studies to assess PPT in the first dorsal interosseous muscle. Force was increased at a rate of 5 N /s. In study 1, two measurements were taken from 240 healthy volunteers (120 males, 120 females; mean age 25 years) giving a power for statistical analysis of beta=0.80 at alpha=0.01. In study two, 30 subjects (15 males, 15 females mean age 28 years) were randomly selected from study one. Fourteen repeated PPT measurements were recorded at seven, 10 min intervals. Mean PPT data for gender groups, from both studies, were analysed using analysis of covariance with repeated measures, and age as the covariate. RESULTS: The mean PPT for each of the two measurements in study one showed a difference between gender of 12.2 N (f=30.5 N, m=42.7 N) and 12.8 N (f=29.5 N, m=42.3 N), respectively, representing a difference of 28% with females exhibiting a lower threshold. In study two, the mean difference calculated from 14 PPT repeated measurements over a 1h period was comparable to that in study one at 12.3N (range 10.4-14.4 N) again females exhibited the lower threshold. The differences in mean PPT values between gender were found to be significant in both study one, at (P<0.0005, F=37.8, df=1) and study two (P=0.01, F=7.6, df=1). No significant differences were found in either study with repeated measurement (P=0.892 and P=0.280), or on the interaction of gender and repeated measurement after controlling for age (P=0.36 and P=0.62). CONCLUSION: Healthy females exhibited significantly lower mean PPTs in the first dorsal interosseous muscle than males, which was maintained for fourteen repeated measures within a 1 h period. This difference is likely to be above clinically relevant levels of change, and it has clear implications for the use of different gender subjects in laboratory based experimental designs utilising PPT as an outcome measure. PMID- 12583869 TI - Reduction of TMD pain by high-frequency vibration: a spatial and temporal analysis. AB - Under some conditions, vibration delivered to the skin can reduce pain (vibratory analgesia). Previous studies of this phenomenon in a clinical context have been somewhat variable in terms of stimulus control, and have not examined the way in which the spatial distribution of pain is affected. In the present study, we used rigorously controlled conditions to examine vibratory analgesia in participants (N=17) with painful temporomandibular disorders (TMD). Results of 20- and 100-Hz vibration were compared with data from a no-vibration control condition. The results document for the first time that vibratory analgesia occurs in TMD chronic pain conditions. We measured its time course using continuous visual analog scale (VAS) recording, and its spatial aspects by asking subjects to indicate painful regions on standardized drawings. VAS ratings and drawings both showed that pain is reduced by 100-Hz, but not by 20-Hz, vibration. The effectiveness of the high-frequency vibration cannot be attributed to a mechanism involving Pacinian corpuscles, since these receptors are lacking in the skin of the orofacial region. Spatial analyses revealed that ipsilateral and contralateral effects of vibration were statistically equivalent, suggesting that vibratory analgesia relies at least in part on central nervous system processes rather than local mechanisms. PMID- 12583870 TI - Effects on muscle pain by intramuscular injection of granisetron in patients with fibromyalgia. AB - We have previously reported that the level of 5-HT in the masseter muscle is increased in patients with fibromyalgia as compared with healthy subjects and that high intramuscular level of 5-HT is associated with muscle pain. We have also reported that injection of the 5-HT(3) receptor antagonist granisetron (GRA) into the masseter muscle of healthy subjects reduced pain induced by 5-HT and abolished allodynia/hyperalgesia. The aim of this study was to investigate whether GRA can influence pain and allodynia/hyperalgesia of the masseter muscle in patients with fibromyalgia. Eighteen female patients who met the criteria of fibromyalgia according to the American College of Rheumatology participated in the study. They were examined regarding pain intensity and pressure pain threshold (PPT) over the masseter muscle. One milliliter of GRA (1 mg/ml) was injected into the masseter muscle on one side and 1 ml of isotonic saline on the other side in a randomized and double-blind manner. After the injections, the pain intensity and PPT were recorded during 30 min. The pain intensity increased after injection of saline and to a lower degree after injection of GRA. The PPT increased after injection of GRA, while no such change was observed after saline. The difference between GRA and saline was, however, not significant. Eight of the patients responded to the GRA injection by an increase of PPT during the experimental period that differed from saline. They also showed a tendency to a lower increase of pain intensity after injection of GRA when compared to saline. In conclusion, the results of this study do not prove that injection of the 5 HT(3)-antagonist GRA into the masseter muscle influences local pain and allodynia/hyperalgesia in patients with fibromyalgia. PMID- 12583871 TI - Differing central amine receptor sensitivity in different migraine subtypes? A neuroendocrine study using buspirone. AB - Despite the importance of the 5HT1A receptor in regulating central serotonergic tone, there is a dearth of research examining its role in migraine. In this study, we examined the hypothesis that there would be altered neuroendocrine responses to a 5HT1A agonist challenge in different migraine subtypes. Prolactin (PRL) responses to the 5HT1A receptor agonist drug buspirone were compared in 30 female subjects with migraine (ten migraine with aura, MA; ten migraine without aura, MO and ten chronic/transformed migraine, CM), and ten healthy controls matched for age, gender and menstrual status. None of the subjects were taking psychotropic medication or migraine prophylactic treatment and those with formal psychiatric disorder were excluded. Endocrine responses were determined by measuring differences between baseline PRL and maximum increases post-buspirone (deltaPRL). There was no difference in baseline PRL between groups. MA subjects did not differ in their PRL responses to buspirone compared to healthy controls. The MO group had a four-fold increase in mean deltaPRL responses compared to healthy controls. Mean deltaPRL was also increased in the CM group compared to controls, but the difference was less exaggerated. This study indicates that there is supersensitive central amine receptor function in MO and CM, but not in MA. These findings support the hypothesis that central 5HT function differs among the migraine subtypes. The results also suggest that migrainous 'transformation' may be associated with adaptive changes in central 5HT receptor sensitivity. The relative contribution of 'state' and 'trait' receptor function to these findings as well as the possible role of dopamine receptors is discussed. PMID- 12583872 TI - Work-related beliefs about injury and physical capability for work in individuals with chronic pain. AB - According to a fear-avoidance model of chronic pain, disability is largely determined by the erroneous belief that an increase in activity level is potentially harmful. Further, recent literature suggests that excessive fears regarding physical activities contribute to significant disability. However, the relation of changes in these fears to functional work capabilities has gone largely uninvestigated. The present study examined how changes in physical capability for work were related to changes in pain severity and fear-avoidance beliefs for general physical and work-specific activities, as well as investigating whether an interdisciplinary treatment program for chronic pain was associated with changes in these specific fears in 65 individuals with chronic pain. Results revealed that significant decreases in fear and pain levels occurred from pre- to post-treatment, in addition to increases in physical capability for work. Further, changes in work-specific fears were more important than changes in pain severity and fear of physical activity in predicting improved physical capability for work. These results expand previous research, which has found a relation between self-reported disability and fear-avoidance beliefs, by demonstrating the relation with fear of work to actual work-related behaviors. PMID- 12583873 TI - The pain vigilance and awareness questionnaire (PVAQ): further psychometric evaluation in fibromyalgia and other chronic pain syndromes. AB - In chronic pain patients, preoccupation with or attention to pain is associated with pain-related fear and perceived pain severity. The current study investigated psychometric properties of the pain vigilance and awareness questionnaire (PVAQ). An exploratory factor analysis on Dutch fibromyalgia patients indicated that a two-factor solution was most suitable. The first factor could be referred to as attention to pain and the second factor was interpreted as attention to changes in pain. A confirmatory factor analysis, testing three different factor structures in two independent samples (Dutch fibromyalgia patients and American pain patients with various diagnoses) showed that the goodness-of-fit indicators for all models were satisfactory. The existence of the previously reported intrusion subscale of the PVAQ as a unique construct within the PVAQ was discussed. This subscale should be further extended by non-reverse keyed items. With regard to the convergent validity, the PVAQ was highly correlated with related constructs such as the pain catastrophizing scale (PCS), pain anxiety symptoms scale (PASS), and Tampa scale of kinesiophobia (TSK). The attention to pain subscale was significantly stronger associated with these pain related measures than the attention to changes in pain subscale, indicating that attention to changes in pain is a distinctive construct. The uniqueness of the attention to changes in pain subscale was also supported by an exploratory factor analysis on all items of the PVAQ, PCS, PASS, and TSK which showed that all items from that scale loaded on one separate factor. Overall, the PVAQ showed good internal consistency. Implications for future research and treatment interventions are discussed. PMID- 12583875 TI - On dialog between clinicians and basic scientists. PMID- 12583876 TI - Ecto-ATPase activity sites in vestibular tissues: an ultracytochemical study in frog semicircular canals. AB - The present study describes the localization and distribution of putative ecto nucleoside-triphosphate-diphosphohydrolases in the frog semicircular canals. These enzymes provide the terminating mechanism of adenosine-5'-triphosphate (ATP) signalling. The localization of the ATP hydrolysis was mapped ultracytochemically using a one-step cerium citrate reaction. Electron-dense precipitates, indicating ecto-adenosine-triphosphatase (ecto-ATPase) activity, were found at the outer surface of plasma membranes of crista hair cells and supporting cells of the sensory epithelium, transitional cells and undifferentiated cells of the ampullar wall and dark cells constituting the secretory epithelium. Non-sensory cells of the ampulla usually exhibited reaction deposits at the level of both apical and basolateral membranes coming into contact with the endolymph and the perilymph respectively, while cells constituting the sensory epithelium showed evident differences in relation to their position. Hair cells and supporting cells of the peripheral regions exhibited clear reaction products both at the level of apical and basolateral membranes, while those of the isthmus region showed abundant reactivity only at the level of their apical membranes. Of particular interest was the observation that hair cell stereocilia exhibited an abundant ecto-ATPase activity, thus suggesting a possible colocalization of enzymatic sites with purinergic receptors and mechanotransduction channels. This strategic expression of ecto-ATPase sites could provide a rapid mechanism of ATP removal able to rapidly restore the sensitivity of transduction channels. In conclusion, the widespread distribution of ecto-ATPase sites at the level of sensory and non-sensory cells of the frog semicircular canals suggests that ATP may have a key role in controlling vestibular function. PMID- 12583877 TI - Gap detection in Mongolian gerbils (Meriones unguiculatus). AB - Hearing thresholds for white-noise stimuli and temporal gap-detection thresholds in six Mongolian gerbils (Meriones unguiculatus) were determined in a GO/NOGO procedure using the method of constant stimuli. Gerbils were offspring of animals collected in the field and only bred in captivity for four generations or less. Hearing thresholds for 800 ms bursts of white noise ranged between -0.8 and 6.3 dB SPL. The median minimum-detectable gap centered in an 800 ms burst of white noise presented at 50 dB SPL was 2.1 ms. At levels of 40 dB SPL and above, gap detection thresholds were independent of the sound-pressure level of the stimulus. At 30 dB SPL and below, the minimum-detectable gap increased with decreasing sound-pressure level. Near 5 dB sensation level, gap-detection thresholds ranged from 13.6 to 29.4 ms. The median threshold for the detection of gaps inserted 5 ms after the onset of an 800 ms burst of white noise of 50 dB SPL was 2.3 ms. The individuals' detection thresholds varied between 2.3 and 3.6 ms for stimuli in which the relative sound-pressure levels of the noise before and after the gap differed by up to 20 dB. The data found in the Mongolian gerbil match results from other mammal and bird species. PMID- 12583878 TI - Myelin-deficiency in the cochlear nerve of the 'bt' mutant hamster. AB - In a previous report, we showed abnormal auditory evoked potentials in the mutant hamster, 'black tremor (bt)', with significantly prolonged wave latencies of auditory brainstem responses and prolonged N1 latencies of compound action potentials, but normal cochlear microphonics. In this report, we present the results of morphological studies supporting the results of our electrophysiological studies of the auditory pathway in bt. Observation by transmission electron microscopy revealed an abnormal myelin sheath surrounding the spiral ganglion cells, and a thinner compact myelin sheath surrounding the axons in bt than in normal hamsters. The bt hamster has a myelin deficiency not only in the brainstem, but also in the cochlear nerve. PMID- 12583879 TI - Spectral dynamics of electroencephalographic activity during auditory information processing. AB - Dynamics of electroencephalographic (EEG) activity during auditory information processing were evaluated in response to changes in stimulus complexity, stimulus discriminability and attention using the oddball paradigm. In comparison to pre stimulus baseline conditions, auditory stimulation synchronized EEG activity in delta, theta and alpha frequency bands. Event-related synchronization (ERS) effects were greatest at approximately 3 Hz (theta frequency band), and their magnitude depended on stimulus and task demands. Event-related desynchronization (ERD) of EEG activity was observed in the beta frequency band. This effect was greatest at approximately 21 Hz but occurred only for easily discriminable stimuli in attention-related target conditions. Because active discrimination tasks also required a button-press response with the right hand, ERDs involved more complex responses that may be related to a combination of perceptual, motor and cognitive processes. These results demonstrate that oddball and attention related EEG responses to auditory stimulation could be characterized in the frequency domain. The specific design and analysis features described herein may prove useful since they provide a simple index of the brain's response to stimulation while at the same time provide powerful information not contained in typical time domain analysis. PMID- 12583880 TI - Primary afferent and cochlear nucleus contributions to extracellular potentials during tone-bursts. AB - Gross electrical responses to tone bursts were measured in the guinea pig with electrodes located in scala tympani (ST) and scala vestibuli (SV) of the cochlea, on the central portion of the VIIIth nerve fibres in the internal auditory meatus, and on the surface of the cochlear nuclear complex (CN). Intracochlear perfusion of pharmacological blockers of neural and postsynaptic activity as well as aspiration of parts or all of the CN were used to dissect the origin of the many components of the gross responses. It was shown that single-ended recordings from either ST or SV or those derived from the sum of the ST and SV responses not only contain mixed responses from the auditory nerve fibres and cochlear hair cells, but are contaminated or modified by neural activity central to the internal auditory meatus, probably in various parts of the CN. Differential recordings between ST and SV were relatively uncontaminated by such activity. Recordings from central locations were largely uncontaminated by potentials from cochlear hair cells. These results suggest that a revised and extended system of nomenclature for the different components of the gross cochlear potentials is necessary, and interpretation of such potentials needs to take into account multiple central as well as peripheral generators. PMID- 12583881 TI - Effect of white noise on vestibular evoked myogenic potentials. AB - OBJECTIVES: To clarify if p13-n23 of vestibular evoked myogenic potentials (VEMPs) is independent of cochlear afferents. METHODS: Twenty normal volunteers and 10 patients with hemifacial palsy enrolled into this study. VEMP and auditory brainstem responses (ABRs) were recorded with or without white noise (WN, 75 dBnHL or 95 dBnHL) ipsilaterally or contralaterally to the stimulated ear. Short tone bursts (STBs) of 0.5 kHz (95 dBnHL, rise/fall time=1 ms, plateau time=2 ms) were presented though headphones. For recording VEMPs surface electromyographic activity was recorded in the supine subjects form symmetrical sites over the upper half of each SCM with a reference electrode over the upper sternum. RESULTS: The amplitude of p13-n23 significantly decreased during exposure of the contralateral ear as well as the ipsilateral ear to 95 dBnHL WN in normal volunteers (41% reduction by the ipsilateral exposure and 38% reduction by the contralateral exposure). Exposure to 75 dBnHL WN caused only minimal reduction of the amplitudes of p13-n23 (5% reduction) although it caused remarkable reduction of the amplitudes of V-SN(10) (ABRs) (23% reduction). However, 95 dBnHL WN exposure showed no effect on the amplitudes of p13-n23 in the ipsilateral side to hemifacial palsy. CONCLUSION: Overall the data are in favor of the hypothesis that cochlear afferents could affect the amplitude of p13-n23 only through the stapedial reflex, although our data might not be strict proof of an absence of influence of cochlear afferents' activity. PMID- 12583882 TI - Spermine modulates neuronal excitability and NMDA receptors in juvenile gerbil auditory thalamus. AB - Medial geniculate body (MGB) neurons process synaptic inputs from auditory cortex. Corticothalamic stimulation evokes glutamatergic excitatory postsynaptic potentials (EPSPs) that vary markedly in amplitude and duration during development. The EPSP decay phase is prolonged during second postnatal week but then shortens, significantly, until adulthood. The EPSP prolongation depends on spermine interactions with a polyamine-sensitive site on receptors for N-methyl-D aspartate (NMDA). We examined effects of spermine application on EPSPs, firing modes, and membrane properties in gerbil MGB neurons during the P14 period of highest polyamine sensitivity. Spermine slowed EPSP decay and promoted firing on EPSPs, without changing passive membrane properties. Spermine increased membrane rectification on depolarization, which is mediated by tetrodotoxin (TTX) sensitive, persistent Na(+) conductance. As a result, spermine lowered threshold and increased tonic firing evoked with current injection by up to approximately 150%. These effects were concentration-dependent (ED(50)=100 microM), reversible, and eliminated by NMDA receptor antagonist, 2-amino-5-phosphonovalerate (APV). In contrast, spermine increased dV/dt of the low threshold Ca(2+) spike (LTS) and burst firing, evoked from hyperpolarized potentials. LTS enhancement was greater at -55 mV than at hyperpolarized potentials and did not result from persistent Na(+) conductance or glutamate receptor mechanisms. In summary, spermine increased excitability by modulating NMDA receptors in juvenile gerbil neurons. PMID- 12583883 TI - Differential changes in Fos-immunoreactivity at the auditory brainstem after chronic injections of salicylate in rats. AB - In human, salicylate-induced tinnitus sometimes occurs a few days after its administration, but the chronic effects of salicylate in animal models are not fully known. In this study, we revealed the distribution of active cells in the rat auditory brainstem by staining an activity marker Fos-protein after multiple daily injections of salicylate. Experimental animals were first given five daily doses of sodium salicylate (250 mg/kg, i.p.). On day 6 they were placed inside a sound room for 8 h before sacrifice. Immunohistochemistry showed a significant increase in the number of Fos-positive cells at the inferior colliculus (IC), particularly its central division. At the cochlear nucleus (CN), only a few Fos stains were found at the dorsal nucleus while no Fos-stain appeared at the ventral nucleus. The scarcity of Fos-stains at the CN reflected more a lack of external sound inputs than an adaptation in Fos-expression. Since Fos-stains in CN could still be induced on day 6 following brief tonal stimulation. Results are consistent with the hypothesis that salicylate-induced tinnitus is a phantom sound perception related to overactivity of cells at the IC. PMID- 12583884 TI - Changes in ultrastructural characteristics of endolymphatic sac ribosome-rich cells of the rat during development. AB - It has recently been demonstrated that endolymphatic sac (ES) ribosome-rich (dark) cells respond to induced endolymph changes and are thus likely to be involved in endolymph homeostasis. Therefore, we studied the ultrastructural characteristics of rat ES ribosome-rich cells during development in order to determine the cellular distribution of organelles involved in protein metabolism, secretion and absorption, indicative for their contribution to endolymph homeostasis. During embryonal stages ribosome-rich cells contain a limited number and variety of organelles and are predominantly involved in the production of components for cell growth and differentiation. In the young adult stage (P60) three different states of ribosome-rich cells may be distinguished. State A resembles a cell with only limited metabolic activities whereas state B is characterized by numerous different intracellular organelles and is considered to be involved in production and secretion as well as absorption and degradation of complex proteins. A third cellular state, state C, is filled with phagolysosomes and contains very few other organelles. This is considered to be a final (pre)apoptotic state. Autoradiography data suggest that ES ribosome-rich cells are capable of synthesis and secretion of tyrosine-containing proteins and may thus be involved in regulation of the osmolarity of endolymph based on the capacity to bind cations as well as water molecules. In addition, ES ribosome rich cells appear to synthesize and secrete fucosylated glycoproteins into the endolymph. In conclusion, the present data suggest that ES ribosome-rich cells are actively involved in endolymph homeostasis through secretion and absorption of complex proteins and it is hypothesized that they are able to adapt their function or activities in response to changes in endolymph composition. PMID- 12583885 TI - Quantification of audiogram fine-structure as a function of hearing threshold. AB - We investigated the amount of fine-structure in the audiograms of patients with Meniere's disease. Both ears (often an affected and an unaffected ear) were investigated. Data are presented from 39 ears of 21 patients with hearing thresholds varying from about 0 to 70 dB HL. The fine-structure was measured in the frequency range from 500 to 3500 Hz; this agrees with a part of the basilar membrane from 10 to 22.5 mm from the apex. The fine-structure was characterized by means of the number of peaks N(p) and the sum S(p) of the heights of all the peaks in an audiogram. From these quantities, we also determined average peak height H(p). We found a negative correlation between hearing loss and strength of fine-structure, i.e. the higher the thresholds the smaller N(p) and S(p) as well as H(p). Also N(p) and H(p) were correlated, i.e. the more peaks the higher the average peak height. The summated peak height S(p)=N(p)H(p) showed a strong dependence on the hearing loss. In cases of strong fine-structure, S(p) reached values around 200 dB. PMID- 12583886 TI - Olivocochlear collaterals evoke excitatory effects in onset neurones of the rat cochlear nucleus. AB - Axons of medial olivocochlear neurones in the superior olivary complex terminate on the outer hair cells of the cochlea and also give off collaterals that terminate in the cochlear nucleus. Previous work in our laboratory, using extracellular recordings in the cochlear nucleus, has indicated that stimulation of the olivocochlear axons may have an excitatory effect on specific cell populations of the cochlear nucleus, such as onset-choppers, in contrast to the peripheral suppressive action of the same axons. We have investigated whether this excitation is produced by action of the olivocochlear collaterals in the cochlear nucleus or whether it is mediated via the peripheral suppression, by measuring intracellular responses in the rat cochlear nucleus to electrical stimulation of the olivocochlear axons in silence. The results demonstrate that single shocks applied to the olivocochlear axons can evoke excitatory postsynaptic potentials in onset neurones. We observed an inhibitory effect in one chopper only. In the same animals in all other neurones investigated (i.e. three primary-like neurones and eight choppers) the same stimulation was without any effect on cell membrane potential. We conclude that the excitatory effects in onset neurones are not caused by suppression in the auditory peripheral organ, but by activation of olivocochlear collaterals in the cochlear nucleus. PMID- 12583887 TI - A novel technique for inducing local inner ear damage. AB - With significant development of mouse genomics and the availability of transgenic and knockout mice, the mouse will be the preferred animal model for inner ear research. However, few studies have used mice as experimental animals for examination of hair cell degeneration, because of their relative resistance to ototoxic agents and difficulties in surgical treatment. This study presents a model for induction of apoptotic cell death in sensory epithelia of the mouse inner ear using injection of neomycin into the posterior semicircular canal. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay revealed that local application of neomycin produced sufficient induction of apoptotic cell death in both auditory and vestibular epithelia over a definite time course. Supplementation of the general caspase-inhibitor significantly reduced induction of TUNEL-positive cells, indicating caspase-dependency of apoptotic cell death observed in the present model. In addition, the approach to the posterior semicircular canal was an easy technique, and sham-operation induced no significant induction of TUNEL-positive cells. This model, hence, enables the use of various genetic tools in studies for mechanisms of hair cell apoptosis. PMID- 12583888 TI - Concurrent measurement of distortion product otoacoustic emissions and auditory steady state evoked potentials. AB - Distortion product otoacoustic emissions (DPOAEs) and auditory steady state evoked response potentials (ASSRs) can both be evoked by tone pairs with frequencies f(1) and f(2). The DPOAE is maximal at 2f(1)-f(2) and the ASSR is maximal at f(2)-f(1). Since DPOAE magnitude depends on the ratio f(2)/f(1), but ASSR amplitude depends on the beat frequency f(2)-f(1), compromises are necessary when recording both responses concurrently. Tone pairs with f(2) of 900, 1800 and 3600 Hz were presented simultaneously at either 40 or 50 dB sound pressure level (SPL). The f(1) frequency of each pair was approximately 85 or 180 Hz lower than f(2). Phase measurements were used to calculate apparent latencies at 40 dB SPL. For increasing f(2), DPOAE latencies were 14.5, 9.7 and 6.3 ms for 85 Hz beats, and 11.5, 9.0 and 4.3 ms for 180 Hz beats. ASSR latencies were 22.0, 15.7 and 17.8 ms at 85 Hz, and 17.7, 11.3 and 9.6 ms at 180 Hz. From a model of the mechanical transmission in the cochlea, delays between the basilar membrane and the generator of the ASSR were estimated as 15.4, 12.2 and 15.3 ms at 85 Hz and 8.6, 7.6 and 8.0 ms at 180 Hz. PMID- 12583890 TI - Giardia lamblia -- a model organism for eukaryotic cell differentiation. AB - Giardia lamblia is a binucleated, flagellated protozoan parasite that inhabits the upper small intestine of its vertebrate hosts. The entire life cycle, which can be completed in vitro, is simple with cycling between a vegetative trophozoite and a highly resistant cystic form. The parasite is one of the earliest diverging eukaryotes known and more than 95% of the genome is sequenced. This makes Giardia an excellent model system for studies of basic eukaryotic processes like cell differentiation. In this review we will discuss recent data concerning Giardia differentiation with a focus on DNA replication and cytokinesis. PMID- 12583891 TI - Trans-translation and protein synthesis inhibitors. AB - Trans-translation is a process found in all bacteria, which contributes to the release of ribosomes that are stalled through a variety of causes, for example when the 3' end of a truncated mRNA lacking a stop codon is reached or at internal clusters of rare codons. Trans-translation requires tmRNA. Trans translation is not essential for cell viability under laboratory conditions, but recently it has been shown that it can contribute to cell viability in the presence of protein synthesis inhibitors. In this minireview, we consider the connection between trans-translation and antibiotics and the potential of using trans-translation as a therapeutic target. PMID- 12583892 TI - Iron-regulated phenylalanyl-tRNA synthetase activity in Azotobacter vinelandii. AB - Azotobacter vinelandii strain UA22 was produced by pTn5luxAB mutagenesis, such that the promoterless luxAB genes were transcribed in an iron-repressible manner. Tn5luxAB was localized to a fragment of chromosomal DNA encoding the thrS, infC, rpmI, rplT, pheS and pheT genes, with Tn5 inserted in the 3'-end of pheS. The isolation of this mutation in an essential gene was possible because of polyploidy in Azotobacter, such that strain UA22 carried both wild-type and mutant alleles of pheS. Phenylalanyl-tRNA synthetase activity and PHES::luxAB reporter activity was partially repressed under iron-sufficient conditions and fully derepressed under iron-limited conditions. The ferric uptake regulator (Fur) bound to a DNA sequence immediately upstream of luxAB, within the pheS gene, but PHES::luxAB reporter activity was not affected by phenylalanine availability. This suggests there is novel regulation of pheST in A. vinelandii by iron availability. PMID- 12583893 TI - The DNA secondary structure of the Bacillus subtilis genome. AB - The entire genomic DNA sequence of the Gram-positive bacterium Bacillus subtilis reported in the SubtiList database has been subjected in this work to a complete bioinformatic analysis of the potential formation of secondary DNA structures such as hairpins and bending. The most significant of these structures have been mapped with respect to their genomic location and compared to those structures already known to have a physiological role, such as the rho-independent transcription terminators. The distribution of these structures along the bacterial chromosome shows two major features: (i). the concentration of the most curved DNA in the intergenic regions rather than within the ORFs, and (ii). a decreasing gradient of large hairpins from the origin towards the terC end of chromosomal DNA replication. Given the increasing biological relevance of secondary DNA structures, these findings should facilitate further studies on the evolution, dynamics and expression of the genetic information stored in bacterial genomes. PMID- 12583894 TI - Identification and disruption of btlA, a locus involved in bile tolerance and general stress resistance in Listeria monocytogenes. AB - A transposon Tn917 mutant of Listeria monocytogenes L028 was isolated on the basis of reduced growth on agar adjusted to pH 5.5. The disrupted gene, designated btlA (bile tolerance locus), encodes a putative secondary transporter of the major facilitator superfamily, which has significant homology to yxiO in Bacillus subtilis (lmo1417 in L. monocytogenes EGDe). The mutant demonstrated decreased growth rates relative to the wild-type when grown in sub-lethal levels of various stressors (acid, salt, ethanol, bile, SDS, ampicillin and phosphomycin). The mutant was also more sensitive to lethal levels of bile. A pORI19 insertion mutant demonstrated similar phenotypes. Murine virulence studies indicated that disruption of btlA does not influence virulence potential. BtlA therefore represents a membrane protein essential for the maintenance of homeostasis under stress conditions, but is not involved in pathogenicity. PMID- 12583895 TI - Conversion of the nitrogen content in liquid manure into biomass and polyglutamic acid by a newly isolated strain of Bacillus licheniformis. AB - Extensive spreading of liquid manure onto agricultural fields causes eutrophication of ground and surface water and also pollution of the atmosphere due to the high ammonium nitrogen content. A poly(gamma-glutamic acid) (PGA) producing strain of Bacillus licheniformis was isolated in this study and investigated for its ability to reduce the ammonium nitrogen by converting ammonium into biomass and PGA as depot forms of nitrogen. In batch cultivations swine manure and an optimized mineral salts medium were used for PGA production. For example the cultivation of B. licheniformis strain S2 in liquid manure, which was modified by adding of 18 g citrate and 80 g glycerol l(-1) and exhibited a carbon to nitrogen ratio of 15.5:1, led to severe reduction of the ammonium content from 2.83 to 0.1 g x l(-1) and to the production of 0.16 g PGA and 7.5 g cell dry mass l(-1) within 410 h. Approximately 28% (w/w) of the total nitrogen was converted into cellular biomass, whereas 0.1% (w/w) was used for the production of PGA. In addition, approximately 33% (w/v) of the original ammonium was lost by stripping. PMID- 12583896 TI - Effect of amino acid starvation on glucose transport in two archaeal organisms. AB - When protein synthesis is arrested by amino acid starvation, Escherichia coli wild-type strains show stringent control (SC) over stable RNA (sRNA) accumulation as well as a large number of other growth-related processes. One of the events under SC is transport of metabolites. Thus, under amino acid starvation, E. coli fails to accumulate the non-metabolizable glucose analog alpha-methyl-D glucoside, whereas isogenic relaxed strains continue to take up this glucose analog. Unlike the Bacteria, most wild-type archaeal strains show relaxed control of sRNA accumulation, although a number of stringent strains have been identified. In order to determine whether stringency in the Archaea affects physiological events different from sRNA accumulation, transport of glucose analogs was examined under amino acid starvation in two stringent archaeal strains, Haloferax volcanii and Sulfolobus acidocaldarius. The experiments were performed with 2-deoxy-D-glucose, which was shown to be transported, but metabolized very limitedly. Unlike E. coli, H. volcanii and S. acidocaldarius continued to transport 2-deoxy-D-glucose under amino acid starvation. Thus, in both Archaea glucose analog transport is not under SC, as it is in E. coli. PMID- 12583897 TI - Optimisation of AP-PCR fingerprinting discriminatory power for clinical isolates of Pseudomonas aeruginosa. AB - Recently methods based on analysis of arbitrarily amplified target sites of microorganism genomes have been extensively applied in microbiological studies. The range of their applications is limited by problems with discrimination and reproducibility resulting from lack of standardised and reliable methods of optimisation. By orthogonal-array optimisation most advantageous and optimal parameters for highly discriminatory primers (CagA2+CMVin2) were selected and efficient AP-PCR (arbitrarily primed-polymerase chain reaction) fingerprinting conditions for Pseudomonas aeruginosa isolates were set up. Stable and multiplex amplicon profiles obtained in this study revealed high level of intraspecies DNA polymorphism among 20 analysed clinical strains of P. aeruginosa proving optimised AP-PCR fingerprinting to be useful in epidemiological typing of the species. PMID- 12583898 TI - Production of D-(--)-3-hydroxyalkanoic acid by recombinant Escherichia coli. AB - Pathways for extracellular production of chiral D-(-)-3-hydroxybutyric acid (3HB) and D-(-)-3-hydroxyalkanoic acid (mcl-3HA) were constructed by co-expression of genes of beta-ketothiolase (phbA), acetoacetyl-CoA reductase (phbB) and 3 hydroxyacyl-ACP CoA transacylase (phaG), respectively, in Escherichia coli strain DH5alpha. The effect of acrylic acid and glucose on production of both 3HB and mcl-3HA was investigated. It was found that the addition of acrylic acid significantly increased production of 3HB and mcl-3HA consisting of 3 hydroxyoctanoic acid and 3-hydroxydecanoic acid in a ratio of 1:3 from 199 mg x l(-1) to 661 mg x l(-1) and from 27 mg x l(-1) to 135 mg x l(-1), respectively, in shake flask studies when glucose was present in the medium at the very beginning of fermentation. The timing of glucose addition had no effect on 3HB production. In contrast, mcl-3HA production was affected by glucose addition, an mcl-3HA concentration of 193 mg x l(-1) was obtained when glucose was added to the culture at 12 h. A more than seven-fold increase was obtained when compared with that in medium containing glucose at the beginning of fermentation. However, a decrease in production of 3HB and mcl-3HA was found when glucose was added at 12 h to the culture containing acrylic acid. The repressive effect of acrylic acid on acetic acid production was also evaluated and discussed. PMID- 12583899 TI - Mutations in sit B and sit D genes affect manganese-growth requirements in Sinorhizobium meliloti. AB - Two transposon-induced mutants of Sinorhizobium meliloti 242 were isolated based on their inability to grow on rich medium supplemented with the metal chelator ethylenediamine di-o-hydroxyphenylacetic acid (EDDHA) and either heme-compounds or siderophores as iron sources. Tagged loci of these mutants were identified as sit B and sit D genes. These genes encode components of an ABC (ATP-binding cassette) metal-type permease in several Gram-negative bacteria. In this work, the phenotypes of these two mutants were compared with those of two siderophore mediated iron transport mutants. The results strongly implicate a role of the sit genes in manganese acquisition when this metal is limiting in S. meliloti. PMID- 12583900 TI - Sucrose accumulation in salt-stressed cells of agp gene deletion-mutant in cyanobacterium Synechocystis sp PCC 6803. AB - The agp gene encoding the ADP-glucose pyrophosphorylase is involved in cyanobacterial glycogen synthesis and glucosylglycerol formation. By in vitro DNA recombination technology, a mutant with partial deletion of agp gene in the cyanobacterium Synechocystis sp. PCC 6803 was constructed. This mutant could not synthesize glycogen or the osmoprotective substance glucosylglycerol. In the mutant cells grown in the medium containing 0.9 M NaCl for 96 h, no glucosylglycerol was detected and the total amount of sucrose was 29 times of that of in wild-type cells. Furthermore, the agp deletion mutant could tolerate up to 0.9 M salt concentration. Our results suggest that sucrose might act as a similar potent osmoprotectant as glucosylglycerol in cyanobacterium Synechocystis sp. PCC 6803. PMID- 12583901 TI - A polymerase chain reaction (PCR) assay specific for Streptococcus suis based on the gene encoding the glutamate dehydrogenase. AB - Polymerase chain reaction (PCR) primers that flank a 688-bp segment within the glutamate dehydrogenase gene (gdh) of Streptococcus suis type 2 could amplify efficiently the DNA of all 306 (100%) clinical S. suis isolates tested (pigs, n=305; human, n=1) encompassing all serotypes obtained from diverse organs, and geographic origins. When DNA from other bacteria were used as templates for amplification, no product was detected indicating specificity of the primers. Multiplex PCR was developed using the gdh gene primer pair and primers that targeted the gene encoding S. suis capsular biosynthesis (cps). This strategy enabled the detection of strains belonging to serotypes 1/2, 1, 2, 7, and 9, respectively. Using the multiplex-PCR technique, 12 out of 14 (86%) isolates that were previously identified as non-typable S. suis (based on biochemical reactions and serology) gave positive PCR results of which four were positive for serotype 7, three for serotype 2, and five for S. suis strains that belong to other serotypes. Retest results of all 14 isolates by several veterinary laboratories were identical with PCR and confirmed that the two non-PCR reactive isolates belonged to strains of other streptococcal species. These results indicated that PCR improved species determination and can thus be used as a reliable species specific molecular diagnostic reagent for the accurate identification of S. suis isolates and a serotype-specific method for the detection of strains of serotypes 1/2, 1, 2, 7, and 9, respectively. The PCR method therefore has potential clinical and epidemiological applications. PMID- 12583902 TI - Identification of peptide inhibitors of Pseudomonas aeruginosa exotoxin A function using a yeast two-hybrid approach. AB - The yeast two-hybrid system was used to identify peptide inhibitors of exotoxin A of Pseudomonas aeruginosa with the goal of using these to design peptide-based drugs against the toxin. A random peptide library consisting of 10(7) peptides ranging in length from 16 to 63 residues was screened for peptides that interact with the C-domain of exotoxin A. From the 10(7) transformants screened, three unique peptides of 63, 61 and 25 amino acids in length were found to specifically interact with the enzyme domain. The genes encoding these peptides were cloned and expressed as fusion proteins with the maltose-binding protein. In vitro inhibition measurements indicated that two of the peptides were modest inhibitors of toxin enzyme activity. These peptides now provide the basis for the development of more potent inhibitors, which will serve as lead inhibitors for evolution of potent peptide-based therapeutics. PMID- 12583903 TI - A novel aldo-keto reductase from Escherichia coli can increase resistance to methylglyoxal toxicity. AB - A novel aldo-keto reductase (AKR) from Escherichia coli has been cloned, expressed and purified. This protein, YghZ, is distantly related (<40%) to mammalian aflatoxin dialdehyde reductases of the aldo-keto reductase AKR7 family and to potassium channel beta-subunits in the AKR6 family. The enzyme has been placed in a new AKR family (AKR14), with the designation AKR14A1. Sequences encoding putative homologues of this enzyme exist in many other bacteria. The enzyme can reduce several aldehyde and diketone substrates, including the toxic metabolite methylglyoxal. The K(m) for the model substrate 4-nitrobenzaldehyde is 1.06 mM and for the endogenous dicarbonyl methylglyoxal it is 3.4 mM. Overexpression of the recombinant enzyme in E. coli leads to increased resistance to methylglyoxal. It is possible that this enzyme plays a role in the metabolism of methylglyoxal, and can influence its levels in vivo. PMID- 12583904 TI - Pectins and pectic-oligosaccharides inhibit Escherichia coli O157:H7 Shiga toxin as directed towards the human colonic cell line HT29. AB - Pectins and pectic-oligosaccharides, as derived by controlled enzymatic hydrolysis, were evaluated for their ability to interfere with the toxicity of Shiga-like toxins from Escherichia coli O157:H7. Both types of material resulted in some degree of protection but this was significantly higher (P>0.01) with the oligosaccharide fractions (giving 90-100% cell survival, compared to 70-80% with the polymer). An effect of methylation on the protective effect was detected with lower degrees being more active. The pectic-oligosaccharides and galabiose, the minimum toxin receptor analogue, were shown to inhibit toxicity and were both protective at 10 mg x ml(-1), but not at lower concentrations. PMID- 12583905 TI - Holin locus characterisation from lysogenic Xenorhabdus nematophila and its involvement in Escherichia coli SheA haemolytic phenotype. AB - Lysogeny has previously been described in the entomopathogenic bacteria of the genus Xenorhabdus. Screening of a X. nematophila prophage DNA library on blood agar resulted in the identification of a 5.7-kb locus that caused a haemolytic phenotype when cloned in Escherichia coli, but not in the E. coli sheA null mutant, lacking the SheA cryptic haemolysin. This locus exhibited similarity to lysis genes from lambdoid phages. In particular, it encoded a functional holin able to complement a lambda Sam7 mutant. It is the second time that a locus encoding a functional holin is shown to reveal the SheA haemolytic phenotype in E. coli. The possible role of the holin in extracellular release of SheA is discussed. PMID- 12583906 TI - Development of a genetic system for hyperthermophilic Archaea: expression of a moderate thermophilic bacterial alcohol dehydrogenase gene in Sulfolobus solfataricus. AB - The Escherichia coli/Sulfolobus solfataricus shuttle vector pEXSs was used as a cloning vehicle for the gene transfer and expression of two bacterial genes in Sulfolobus solfataricus. The alcohol dehydrogenase (adh) from the moderate thermophilic Bacillus stearothermophilus (strain LLDR) and a mutagenised version encoding a less thermostable ADH enzyme were the selected genes. S. solfataricus adh promoter and aspartate aminotransferase terminator were used to drive the heterologous gene expression and to guarantee the correct termination of the transcripts, respectively. The constructed vectors were found to be able to carry these 'passenger' genes without undergoing any rearrangements. The active transcription of bacillar mRNAs was ascertained in vivo by RT-PCR. Transformed S. solfataricus expressed functional exogenous ADHs that showed unaffected kinetic and chemical-physical features. PMID- 12583907 TI - Identification and characterization of KvgAS, a two-component system in Klebsiella pneumoniae CG43. AB - A two-component system encoding gene cluster kvgAS that is present only in virulent Klebsiella pneumoniae CG43 was isolated and its sequence determined. RT PCR and Southern analysis demonstrated that kvgAS is organized as an operon. No apparent effect of a kvgS deletion on bacterial virulence was observed in a mouse peritonitis model. In the presence of paraquat or 2,2-dipyridyl, the activity of kvgAS promoter in the kvgS mutant was found to be reduced to half of the level in the wild-type strain. The data suggest that the KvgAS system is autoregulated and plays a role in countering free radical stresses and sensing iron-limiting conditions. PMID- 12583908 TI - Growth and colonization suppression of Salmonella enterica serovar Hadar in vitro and in vivo. AB - Growth suppression in Salmonella enterica serovar Hadar (S. Hadar) was investigated, in vitro under strict anaerobiosis and in vivo in the intestine of the day-old chicken. Stationary-phase cultures of 20 S. Hadar field strains were tested against each other for growth suppression activity by their ability to suppress the multiplication of low counts of minority cultures inoculated into them as nalidixic acid-resistant mutants. All strains showed profound growth suppression. Four S. Hadar strains were selected and further tested for their ability to suppress growth of S. Enteritidis, S. Typhimurium, S. Virchow and S. Saintpaul. One of the four strains (S. Hadar 18) was randomly selected for further studies. Precolonization of chicken with S. Hadar 18 prevented superinfection with any of the serovars mentioned above. From more than 1000 TnphoA mutants of S. Hadar 18 screened against the parent strain anaerobically in vitro, four were non-suppressive with TnphoA insertions in dapF, aroD, sgaT or tatA. Only the dapF mutant was also non-suppressive in the chicken intestine. PMID- 12583909 TI - Organization of the biosynthetic gene cluster for the polyketide macrolide mycinamicin in Micromonospora griseorubida. AB - Mycinamicin, composed of a branched lactone and two sugars, desosamine and mycinose, at the C-5 and C-21 positions, is a 16-membered macrolide antibiotic produced by Micromonospora griseorubida A11725, which shows strong antimicrobial activity against Gram-positive bacteria. The nucleotide sequence (62 kb) of the mycinamicin biosynthetic gene cluster, in which there were 22 open reading frames (ORFs), was completely determined. All of the products from the 22 ORFs are responsible for the biosynthesis of mycinamicin II and self-protection against the compounds synthesized. Central to the cluster is a polyketide synthase locus (mycA), which encodes a seven-module system comprised of five multifunctional proteins. Immediately downstream of mycA, there is a set of genes for desosamine biosynthesis (mydA-G and mycB). Moreover, mydH, whose product is responsible for the biosynthesis of mycinose, lies between mydA and B. On the other hand, eight ORFs were detected upstream of the mycinamicin PKS gene. The myrB, mycG, and mycF genes had already been characterized by Inouye et al. The other five ORFs (mycCI, mycCII, mydI, mycE, and mycD) lie between mycA1 and mycF, and these five genes and mycF are responsible for the biosynthesis of mycinose. In the PKS gene, four regions of KS and AT domains in modules 1, 4, 5, and 6 indicated that it does not show the high GC content typical for Streptomyces genes, nor the unusual frame plot patterns for Streptomyces genes. Methylmalonyl-CoA was used as substrate in the functional units of those four modules. The relationship between the substrate and the unusual frame plot pattern of the KS and AT domains was observed in the other PKS genes, and it is suggested that the KS-AT original region was horizontally transferred into the PKS genes on the chromosomal DNA of several actinomycetes strains. PMID- 12583910 TI - Induction of laccase activity in the edible straw mushroom, Volvariella volvacea. AB - Volvariella volvacea, strain V14, produces multiple forms of extracellular laccase when grown in submerged culture in a defined medium with glucose as sole carbon source, and on cotton waste 'compost' representative of the conditions used for industrial-scale mushroom cultivation. In liquid culture, enzyme synthesis is associated with the onset of secondary growth, and is positively regulated by copper (up to 200 microM CuSO(4)) and by various aromatic compounds. In solid-state systems, only low levels of laccase are detectable during the vegetative growth phase but enzyme activity increases sharply at the onset of fruiting and during sporophore development. PMID- 12583911 TI - Role of capsule in Klebsiella pneumoniae virulence: lack of correlation between in vitro and in vivo studies. AB - In vitro and in vivo models were used to investigate the role of capsule on the virulence of Klebsiella pneumoniae. We showed that capsule expression reduces dramatically the ability of the K. pneumoniae to bind to epithelial cells when compared to its non-capsulated variant. The presence/absence of capsule had no effect on the colonization of the gastrointestinal tract, while in the urinary tract we established that capsule is an important virulence factor. Our study demonstrates the caution needed when extrapolating from results of in vitro studies and emphasizes the necessity of in vivo models in studies of bacterial virulence. PMID- 12583912 TI - Different cellular fatty acid pattern behaviours of two strains of Listeria monocytogenes Scott A and CNL 895807 under different temperature and salinity conditions. AB - Cells of two strains of Listeria monocytogenes CNL 895807 and Scott A were grown to late exponential phase at different growth temperatures (37, 20 and 4 degrees C) with or without NaCl (7%), and their fatty acid compositions were analysed. The results showed that low thermal adaptation response of L. monocytogenes CNL was different than that of the Scott A strain, and it was based on both an increase of anteiso-branched-chain fatty acids and a significant decrease of straight-chain fatty acids. However, the main modifications observed in the Scott A strain when grown at a low temperature were a decrease of the proportion of ai17:0 and an increase of ai15:0. In hyperosmotic medium and over the entire temperature range (4 degrees C, 20 degrees C and 37 degrees C) the two L. monocytogenes strains showed a cellular fatty acid profile dominated by ai15:0. In addition, a decrease of the two major straight-chain fatty acids (14:0 and 16:0) was observed in the CNL strain. These results demonstrated that the CNL strain showed different behaviours of low thermal and salt adaptation to maintain membrane fluidity, which are based both on an increase of anteiso-branched-chain fatty acids, and a significant decrease of straight-chain fatty acids. PMID- 12583913 TI - Microorganisms cultured from stratospheric air samples obtained at 41 km. AB - Samples of air removed from the stratosphere, at an altitude of 41 km, were previously found to contain viable, but non-cultureable bacteria (cocci and rods). Here, we describe experiments aimed at growing these, together with any other organisms, present in these samples. Two bacteria (Bacillus simplex and Staphylococcus pasteuri) and a single fungus, Engyodontium album (Limber) de Hoog were isolated from the samples. Although the possibility of contamination can never be ruled out when space-derived samples are studied on earth, we are confident that the organisms originated from the stratosphere. Possible mechanisms by which these organisms could have attained such a height are discussed. PMID- 12583914 TI - Isolation and phylogenetic diversity of members of previously uncultivated epsilon-Proteobacteria in deep-sea hydrothermal fields. AB - We report the successful cultivation and partial characterization of novel members of epsilon-Proteobacteria, which have long been recognized solely as genetic signatures of small subunit ribosomal RNA genes (rDNA) from a variety of habitats occurring in deep-sea hydrothermal fields. A newly designed microhabitat designated 'in situ colonization system' was used for enrichment. Based on phylogenetic analysis of the rDNA of the isolates, most of these represent the first cultivated members harboring previously uncultivated phylotypes classified into the Uncultivated epsilon-Proteobacteria Groups A, B, F and G, as well as some novel members of Group D. Preliminary characterization of the isolates indicates that all are mesophilic or thermophilic chemolithoautotrophs using H(2) or reduced sulfur compounds (elemental sulfur or thiosulfate) as an electron donor and O(2), nitrate or elemental sulfur as an electron acceptor. The successful cultivation will enable the subsequent characterization of physiological properties and ecological impacts of a diversity of epsilon Proteobacteria in the deep-sea hydrothermal environments. PMID- 12583915 TI - The localization change of Ybr078w/Ecm33, a yeast GPI-associated protein, from the plasma membrane to the cell wall, affecting the cellular function. AB - The YBR078W/ECM33 gene of Saccharomyces cerevisiae encodes a glycosylphosphatidylinositol (GPI)-attached protein and its disruptant strain exhibited a temperature-sensitive (ts) growth defect. A HA-tagged Ybr078w protein, which complemented the ts growth phenotype of the ybr078wdelta strain, was predominantly located on the plasma membrane by GPI anchoring. To examine the requirement of the GPI anchoring on the plasma membrane for the function, the omega-minus region of Ybr078w was replaced with those of Ydr534c/Fit1 and Ynl327w/Egt2, which are known as GPI-dependent cell wall proteins. The replacement induced the change in localization of the mutant proteins from the plasma membrane to the cell wall and the mutant proteins lost the function to complement the ts cell growth defect of the ybr078wdelta strain. In addition, a similar result was obtained in a mutant protein, where the authentic SKKSK sequence at the omega-5 to omega-1 site of Ybr078w was replaced with a synthetic ISSYS sequence. It is concluded that the GPI anchoring on the plasma membrane is required for the Ybr078w function. PMID- 12583916 TI - Artificial chromosome libraries of Streptomyces coelicolor A3(2) and Planobispora rosea. AB - Using an Escherichia coli-Streptomyces shuttle vector derived from a bacterial artificial chromosome (BAC), we developed methodologies for the construction of BAC libraries of filamentous actinomycetes. Libraries of Streptomyces coelicolor, the model actinomycete, and Planobispora rosea, a genetically intractable strain, were constructed. Both libraries have an average insert size of 60 kb, with maximal insert larger than 150 kb. The S. coelicolor library was evaluated by selected hybridisations to DraI fragments and by end sequencing of a few clones. Hybridisation of the P. rosea library to selected probes indicates a good representation of the P. rosea genome and that the library can be used to facilitate the genomic analysis of this actinomycete. PMID- 12583917 TI - In vivo detection of molybdate-binding proteins using a competition assay with ModE in Escherichia coli. AB - Molybdenum is an important trace element as it forms the essential part of the active site in all molybdenum-containing enzymes. We have designed an assay for the in vivo detection of molybdate binding to proteins in Escherichia coli. The assay is based on (i). the molybdate-dependent transcriptional regulation of the moa operon by the ModE protein, and (ii). the competition for molybdate between ModE and other molybdate-binding proteins in the cytoplasm of E. coli. We were able to verify in vivo molybdate binding to three different bacterial proteins that are known to bind molybdate. This sensitive in vivo system allows the testing of different proteins for molybdate binding under in vivo conditions and will facilitate the identification of other cellular factors needed for molybdate binding. As a first example, we examined the eukaryotic protein Cnx1 that is involved in the last step of molybdenum cofactor biosynthesis in plants, and show that it is able to compete with ModE for molybdate in a molybdopterin-dependent fashion. PMID- 12583918 TI - MYO2 is not essential for viability, but is required for polarized growth and dimorphic switches in Candida albicans. AB - The human fungal pathogen Candida albicans changes from a budding yeast form to a polarized hyphal form in response to various external conditions. Dimorphic switching of C. albicans has been implicated in the development of pathogenicity. Morphogenic transformation requires polarized cell growth and rearrangement of the cytoskeleton. We previously showed that myosins play key roles in the conversion from the bud to the hyphal form of C. albicans by inhibiting myosin activities with 2,3-butanedione-2-monoxime (BDM), a general myosin ATPase inhibitor. In this study we investigated the function of MYO2 in C. albicans using deletion mutants. The amino acid sequence of CaMYO2 shows 60% identity and 77% homology with MYO2 and 54% identity and 70% homology with MYO4 of budding yeast Saccharomyces cerevisiae, suggesting that CaMYO2 is the only class V myosin in C. albicans. Cells in which both CaMYO2 alleles were deleted were viable, suggesting that MYO2 is nonessential in C. albicans. The proliferation of CaMYO2delta cells, however, was sharply decreased. In addition, CaMYO2delta cells showed defects in assembly and polarized localization of F-actin as well as an inability to induce germ tube formation and hyphal growth. The deletion of CaMYO2 also disrupted the shape and migration of the nucleus. These results strongly suggest that CaMYO2 is essential for polarized growth and hyphal transition in C. albicans. PMID- 12583919 TI - Occurrence of Salmonella spp and Cryptosporidium spp in a French coastal watershed: relationship with fecal indicators. AB - Cryptosporidium and Salmonella are pathogenic microorganisms that can cause severe gastrointestinal illness in humans. Because these organisms are potentially transmitted through natural waters, this study was carried out to estimate the concentrations of both pathogens in a French coastal watershed and to determine the relationships with fecal indicators. Water samples from nine wastewater treatment plants and eight rivers were analyzed. Although both pathogens and indicators are released from sewage effluents, no clear correlation was found between the two enteric pathogens nor between a given pathogen and fecal indicators. These results suggest that fecal indicators do not adequately indicate the presence of Cryptosporidium and Salmonella in natural waters and that pathogens and indicators may have different behaviors in the aquatic environment. PMID- 12583920 TI - The somatization in primary care study: a tale of three diagnoses. AB - Somatization is a common phenomenon that has been defined in many ways. The two most widely used diagnoses, Somatization Disorder (SD) and Abridged Somatization Disorder (ASD), are based on lifetime unexplained symptoms. However, reports indicate instability in lifetime symptom recall among somatizing patients. Multisomatoform disorder (MSD) is a new diagnosis based on current unexplained symptoms. To understand how knowledge about SD and ASD translates to MSD, we examined the diagnostic concordance, impairment and health care utilization of these groups in a sample from the Somatization in Primary Care Study. The diagnostic concordance was high between MSD and SD, but lower between MSD and ASD. All three groups reported considerable physical impairment (measured using the PCS subscale of the SF-36). The mental health (MCS) scores for the three groups were only slightly lower than those of the general population. Over the course of one year, physical functioning fell significantly for all three groups. Mental functioning did not change significantly for any of the three groups over this period. Utilization patterns were very similar for the three groups. The high prevalence, serious impairment, and worsening physical functioning over the course of one year suggest the importance of developing interventions in primary care to alleviate the impaired physical functioning and reduce utilization in somatizing patients. MSD should be a useful diagnosis for targeting these interventions because it identifies a sizable cohort of somatizing patients reporting impairment of comparable severity to full SD, using a more efficient diagnostic algorithm based on current symptoms. PMID- 12583921 TI - Somatoform disorders in consultation-liaison psychiatry: a comparison with other mental disorders. AB - Consultation-liaison (C-L) psychiatry has an important role in the management of somatoform disorders (SD). Characteristics of SD patients in C-L psychiatry are largely unknown and are presented in this paper. We analyzed 13,314 Dutch psychiatric consultations from 1984 to 1991 and compared patients diagnosed with SD to patients with other mental disorders and to those without a mental disorder. The comparison included socio-demographic variables, consult characteristics, medical history, current somatic morbidity, information about additional diagnostic tests, hospital admission time and aftercare management. Of the 544 SD patients 39.5% (n = 215) were diagnosed with a conversion disorder that illustrates the highly selected nature of SD patients in C-L psychiatry. Employment among SD patients decreased significantly from 58% in the group aged 20-29 years to 6% in the group aged 50-59 years. This decrease was significantly larger as compared to other mental disorders and no mental disorders and was virtually unaffected by correction for potential confounding by gender. Contrary to our expectation no difference between the three groups was observed in claims for disability benefits. Of the SD patients 74.5% were referred for aftercare management, significantly more than the other two groups which is considered a promising development in C-L psychiatry. PMID- 12583922 TI - The impact of psychiatric comorbidity on length of stay of medical inpatients. AB - The aim of the study was to determine the impact of psychiatric comorbidity (PC) on length of hospital stay (LOS) of medical inpatients. A prospective cohort study was conducted. A series of 317 medical inpatients consecutively admitted to the general medical wards of a University Hospital composed the sample, after excluding those who refused or who could not be evaluated due to their physical illnesses or treatments (n=78). Data on demographic and medical variables were collected. A psychiatrist categorized subjects into two cohorts (with and without PC), according to DSM-IV, using the Schedule for Affective Disorders and Schizophrenia, except in patients cognitively impaired who were diagnosed by clinical interview. Mortality and length of stay during the index hospitalization were recorded. At admission, 156 (49%) inpatients had a current psychiatric comorbidity. After controlling for confounders (age and physical severity), in the multivariate analysis of covariance, the patients with cognitive impairment had a significantly prolonged LOS (F=17.8; P<.01) compared with those without cognitive impairment. No difference existed in LOS for the patients with depressive disorders (F=0.36; P=.55), Anxiety disorders (F=1.48; P=.22) or Substance related disorders (F=1.05; P=.30). These results suggest an independent effect of cognitive impairment increasing LOS of medical inpatients. PMID- 12583923 TI - The consultation psychiatrist as effective physician. AB - The two fundamental tasks of the consultation psychiatrist, diagnosis and treatment, can sometimes seem at odds with the consultee's desired plan or outcome. While implicitly recognized, this fact is rarely explicitly taught to residents or addressed in the literature. The authors proffer three principles of inpatient consultation psychiatry aimed at dealing with tensions that can arise over issues of; 1) diagnosis (Principle I: Do not be afraid to refute notions of psychopathology); 2) medication (Principle II: Do not be afraid to discontinue unnecessary psychotropics); and 3) responsibility (Principle III: Do not be afraid to "sign off" when necessary). These principles emphasize the medical model, the status of the psychiatrist as a medical expert, and the importance of defining one's professional limits. PMID- 12583924 TI - Use of herbal medications among 200 psychiatric outpatients: prevalence, patterns of use, and potential dangers. AB - We documented the use rates of herbal medication among a population of psychiatric outpatients to identify patients at risk for drug-herb interactions. Data were collected on 200 patients who, as part of their routine evaluation, provided information regarding their use of herbs. Fifteen percent of patients were currently taking herbs, the majority for treatment of psychiatric symptoms. There were no significant differences in use patterns across gender or between age groups. None of the current users charts, including the "high risk" patients, indicated that treating physicians were aware of potentially dangerous drug-herb combinations. Use of herbal medication was common among this sample of psychiatric outpatients. These data are consistent with reports from other patient populations. It is important for psychiatrists to ask all patients about their use of herbs and provide education about potentially dangerous drug-herb interactions. PMID- 12583925 TI - Steroid psychosis: a review. AB - This review is built upon a time-framed perspective to unfold the growth of evidence and the shifting of focus from primary affective short-term reactions to later findings of cognitive deficits and possible permanent impairment linked to steroid treatment. An incidence related to dosage has been documented and delirium and withdrawal symptoms have been reported in later studies. A hypothesis of sensitization process with multiple course of steroids has been proposed with the reporting of recurrent cases. The issue of individual risk appears unsettled while management of psychiatric reactions to steroids has shifted toward prophylactic use of lithium. PMID- 12583926 TI - Incidence and clinical course of major depression in patients with chronic hepatitis type C undergoing interferon-alpha therapy: a prospective study. AB - This study examined the incidence, clinical course and its risk factors for major depression in patients with chronic hepatitis type C undergoing interferon-alpha therapy. Ninety-nine subjects underwent the psychiatric interviews for diagnosis of major depressive episode according to the DSM-IV criteria before the start of interferon therapy, and once every 4 weeks during both the 24-week treatment period and 12 weeks after the end of therapy. Depressive symptoms were also evaluated using the Hamilton Rating Scale for Depression. Major depression occurred during interferon therapy in 23 patients (23.2%). In 73.9% of them depression occurred within 8 weeks after the start of therapy. Twenty-two patients with depression completed the therapy and 59.1% of them achieved remission by the end of therapy with a mean duration of 11.6 weeks. Although the other 40.9% were not in remission at the end of therapy, they achieved remission within 12 weeks thereafter. The only risk factor for depression was advanced age. Depression occurs frequently among patients with hepatitis type C undergoing interferon-alpha therapy. Such patients require careful observation, and psychiatrists should be sufficiently aware of this significant psychiatric complication of interferon therapy. PMID- 12583927 TI - Acute-phase treatment in general hospitals: clinical psychopharmacologic evaluation in first-episode schizophrenia patients. AB - A case-control study was conducted to investigate possible predictors of response to antipsychotic treatment in acute schizophrenic patients. During a 14-month period, 13 female first-episode schizophrenic patients were included. According to their antipsychotic treatment response 8 weeks after admission, they were placed in either a good-response group (n = 8) or poor-response group (n = 5). Scores of the two groups differed on several items of the Positive and Negative Syndrome Scale (PANSS) at baseline and during the following 8 weeks. At baseline, the maximum scores for Conceptual disorganization, Poor rapport, Passive/apathetic social withdrawal, and Stereotyped thinking in the good response group were lower than the corresponding minimum scores in the poor response group. Furthermore, minimum improvements after 1 week in scores for excitement in the good-response group were higher than the maximum improvements in the corresponding scores of the poor-response group. These PANSS items might serve as simple and rapid predictors of treatment resistance. In addition, Lack of judgment and insight significantly improved by 1 week, and further improved by 5 weeks in the good-response group. This suggests that treatment in the good response group will be optimal if provided during a hospital stay of appropriate length. PMID- 12583928 TI - Discharge against medical advice at a general hospital in Catalonia. AB - Some studies on discharge against medical advice (AMA) in general hospitals report a prevalence between 0.7-7% with 11-42% of this population identified as psychiatric patients. To study the sociodemographic and psychopathological features of patients who leave AMA, we performed a retrospective case-control comparison study of length of hospitalization and presence of psychiatric disturbances on patients who left AMA from the University General Hospital in Catalan Spain over a two-year period. An analysis of the hospital epidemiological discharge register and retrospective chart review for presence of psychiatric disturbances found that AMA prevalence was 0.34%, the total discharge number in the 2-year period being 41,648. AMA rates by medical department were 0.44% for the internal medicine department; 0.24% for surgery; 0.26% for orthopedic surgery, 0.32% for obstetrics-gynecology and 0.93% for rehabilitation. The mean age for AMA patients was 38.63 years, with a higher number of men (59.9%). A total of 45.8% AMA discharges were from the internal medicine department. No significant differences were found in the average length of hospitalization between the AMA and control groups. The presence of psychiatric pathology was significantly higher among the AMA group (P<.05). The prevalence of AMA at our hospital was low in comparison to the rates reported in the literature. The patient at high risk for AMA discharge is a young man with a history of psychiatric pathology, mainly narcotic dependence. PMID- 12583929 TI - Paroxetine-induced limb anesthesia. PMID- 12583930 TI - Relationship between anxiety/depression and nausea: causal or associative? PMID- 12583932 TI - A patient with lithium intoxication developing at therapeutic serum lithium levels and persistent delirium after discontinuation of its administration. PMID- 12583933 TI - Postictal psychosis coexisting with forced thinking. PMID- 12583934 TI - Clonazepam augmentation therapy in a male at early adolescence with rapid cycling bipolar disorder. PMID- 12583935 TI - Venous thromboembolism and clozapine: an association unresolved. PMID- 12583936 TI - The neglect of child neglect. PMID- 12583937 TI - COPD: treatments benefit patients. PMID- 12583938 TI - Revisiting the use of the electronic fetal monitor. PMID- 12583939 TI - Child sexual abuse: vicious circles of fate or paths to resilience? PMID- 12583940 TI - Genetic susceptibility to variant Creutzfeldt-Jakob disease. PMID- 12583941 TI - Drugs and falls in later life. PMID- 12583943 TI - The right word. PMID- 12583942 TI - Combined salmeterol and fluticasone in the treatment of chronic obstructive pulmonary disease: a randomised controlled trial. AB - BACKGROUND: Inhaled long-acting beta2 agonists improve lung function and health status in symptomatic chronic obstructive pulmonary disease (COPD), whereas inhaled corticosteroids reduce the frequency of acute episodes of symptom exacerbation and delay deterioration in health status. We postulated that a combination of these treatments would be better than each component used alone. METHODS: 1465 patients with COPD were recruited from outpatient departments in 25 countries. They were treated in a randomised, double-blind, parallel-group, placebo-controlled study with either 50 microg salmeterol twice daily (n=372), 500 microg fluticasone twice daily (n=374), 50 microg salmeterol and 500 microg fluticasone twice daily (n=358), or placebo (n=361) for 12 months. The primary outcome was the pretreatment forced expiratory volume in 1s (FEV1) after 12 months treatment' and after patients had abstained from all bronchodilators for at least 6h and from study medication for at least 12h. Secondary outcomes were other lung function measurements, symptoms and rescue treatment use, the number of exacerbations, patient withdrawals, and disease-specific health status. We assessed adverse events, serum cortisol concentrations, skin bruising, and electrocardiograms. Analysis was as predefined in the study protocol. FINDINGS: All active treatments improved lung function, symptoms, and health status and reduced use of rescue medication and frequency of exacerbations. Combination therapy improved pretreatment FEV1 significantly more than did placebo (treatment difference 133 mL, 95% CI 105-161, p<0.0001), salmeterol (73 mL, 46-101, p<0.0001), or fluticasone alone (95 mL, 67-122, p<0.0001). Combination treatment produced a clinically significant improvement in health status and the greatest reduction in daily symptoms. All treatments were well tolerated with no difference in the frequency of adverse events, bruising, or clinically significant falls in serum cortisol concentration. INTERPRETATION: Because inhaled long-acting beta2 agonists and corticosteroid combination treatment produces better control of symptoms and lung function, with no greater risk of side-effects than that with use of either component alone, this combination treatment should be considered for patients with COPD. PMID- 12583944 TI - Comparison of intermittent and continuous palliative chemotherapy for advanced colorectal cancer: a multicentre randomised trial. AB - BACKGROUND: Policies of UK clinicians regarding the duration of chemotherapy for patients with advanced colorectal cancer are not consistent. We aimed to compare effectiveness of continuous and intermittent chemotherapy in such patients. METHODS: Patients who responded or had stable disease after receiving 12 weeks of the regimens described by de Gramont and Lokich, or raltitrexed chemotherapy, were randomised to either intermittent (a break in chemotherapy, re-starting on the same drug on progression), or continuous chemotherapy until progression. FINDINGS: 354 patients (178 intermittent, 176 continuous) were enrolled from 42 UK centres. At randomisation, 41% of participants had part or complete response; 59% were stable. Only 66 (37%) patients allocated to intermittent treatment restarted as planned, after a median of 130 days. Median time on treatment after restarting was 84 days. Patients in the continuous group remained on treatment for a median of a further 92 days. Similar proportions of patients in both groups received second-line therapy. Patients on intermittent chemotherapy had significantly fewer toxic effects and serious adverse events than those in the continuous group. There was no clear evidence of a difference in overall survival (hazard ratio 0.87 favouring intermittent, 95% CI 0.69-1.09, p=0.23). INTERPRETATION: Our findings provided no clear evidence of a benefit in continuing therapy indefinitely until disease progression. They showed that it is safe to stop chemotherapy after 12 weeks and re-start the same treatment on progression in patients with chemosensitive advanced colorectal cancer. PMID- 12583945 TI - Admission cardiotocography: a randomised controlled trial. AB - BACKGROUND: Admission cardiotocography is widely used to identify pregnancies that might benefit from continuous electronic fetal monitoring in labour. We aimed to compare the effect on neonatal outcome of admission cardiotocography versus intermittent auscultation of the fetal heart rate. METHODS: 8580 women admitted to the delivery ward of a Dublin teaching hospital who were at low risk of fetal distress in labour were randomly assigned admission cardiotocography (20 min) or the unit's usual care (intermittent auscultation only, with continuous cardiotocography only if clinically indicated). The primary outcome was moderate to severe neonatal morbidity, or perinatal mortality in the absence of a major congenital malformation. Analyses were by intention to treat. FINDINGS: 44 (1.0%) women assigned admission cardiotocography did not undergo the procedure; 15 (0.4%) assigned usual care had admission cardiotocography. The primary endpoint occurred in 56 (1.3%) of 4298 women assigned admission cardiotocography and 55 (1.3%) of 4282 in the usual-care group (relative risk 1.01; 95% CI 0.70-1.47). Other indices of neonatal morbidity also showed no differences. Despite an increase in use of continuous cardiotocography (1.39; 1.33-1.45) and fetal blood sampling (1.30; 1.14-1.47) with admission cardiotocography, there were no significant differences in the rates of caesarean delivery (1.13; 0.92-1.40), instrumental delivery (1.03; 0.92-1.16), or episiotomy (1.06; 0.99-1.13). INTERPRETATION: Routine use of cardiotocography for 20 min on admission to the delivery ward does not improve neonatal outcome. No significant increase in operative delivery was apparent, probably because of liberal use of fetal blood sampling. PMID- 12583946 TI - Development of sexually abusive behaviour in sexually victimised males: a longitudinal study. AB - BACKGROUND: Sexual maltreatment is one of the most common forms of child abuse. To identify risk factors for sexually abusive behaviour by adults, we prospectively assessed childhood experiences and personal characteristics of male child victims who became abusers in later life. METHODS: In a longitudinal study (7-19 years duration), we included 224 former male victims of sexual abuse. Risk factors contemporaneous with the abuse, and putative protective influences, were identified from social service and clinical records. Evidence of later criminal acts was obtained from a nationwide search of official records. FINDINGS: Of the 224 former victims, 26 had subsequently committed sexual offences (victim abusers), in almost all cases with children, mainly outside their families. Risk factors during childhood for later offending included material neglect (odds ratio 3.4, 95% CI 1.2-9.7), lack of supervision (3.0, 1.1-8.3), and sexual abuse by a female person (3.0, 1.1-8.7). Victim-abusers had more frequently witnessed serious intrafamilial violence (3.1, 1.0-10.0). Six (29%) of 21 victim-abusers on whom we had relevant data had been cruel to animals (7.9, 2.0-31.4). No single putative protective factor, nor a composite protective index, significantly reduced the risk of paedophilic behaviour. INTERPRETATION: Most male victims of child sexual abuse do not become paedophiles, but particular experiences and patterns of childhood behaviour are associated with an increased risk of victims becoming abusers in later life. Our findings have implications for the design of selective interventions with a vulnerable subgroup of male victims, aimed at reducing the risk of paedophilic behaviour in later life. PMID- 12583947 TI - Association of n-3 polyunsaturated fatty acids with stability of atherosclerotic plaques: a randomised controlled trial. AB - BACKGROUND: N-3 polyunsaturated fatty acids (PUFAs) from oily fish protect against death from cardiovascular disease. We aimed to assess the hypothesis that incorporation of n-3 and n-6 PUFAs into advanced atherosclerotic plaques increases and decreases plaque stability, respectively. METHODS: We did a randomised controlled trial of patients awaiting carotid endarterectomy. We randomly allocated patients control, sunflower oil (n-6), or fish-oil (n-3) capsules until surgery. Primary outcome was plaque morphology indicative of stability or instability, and outcome measures were concentrations of EPA, DHA, and linoleic acid in carotid plaques; plaque morphology; and presence of macrophages in plaques. Analysis was per protocol. FINDINGS: 188 patients were enrolled and randomised; 18 withdrew and eight were excluded. Duration of oil treatment was 7-189 days (median 42) and did not differ between groups. The proportions of EPA and DHA were higher in carotid plaque fractions in patients receiving fish oil compared with those receiving control (absolute difference 0.5 [95% CI 0.3-0.7], 0.4 [0.1-0.6], and 0.2 [0.1-0.4] g/100 g total fatty acids for EPA; and 0.3 [0.0-0.8], 0.4 [0.1-0.7], and 0.3 [0.1-0.6] g/100 g total fatty acids for DHA; in plaque phospholipids, cholesteryl esters, and triacylglycerols, respectively). Sunflower oil had little effect on the fatty acid composition of lipid fractions. Fewer plaques from patients being treated with fish oil had thin fibrous caps and signs of inflammation and more plaques had thick fibrous caps and no signs of inflammation, compared with plaques in patients in the control and sunflower oil groups (odds ratio 0.52 [95% CI 0.24-0.89] and 1.19 [1.02-1.57] vs control; 0.49 [0.23-0.90] and 1.16 [1.01-1.53] vs sunflower oil). The number of macrophages in plaques from patients receiving fish oil was lower than in the other two groups. Carotid plaque morphology and infiltration by macrophages did not differ between control and sunflower oil groups. INTERPRETATION: Atherosclerotic plaques readily incorporate n-3 PUFAs from fish-oil supplementation, inducing changes that can enhance stability of atherosclerotic plaques. By contrast, increased consumption of n-6 PUFAs does not affect carotid plaque fatty-acid composition or stability over the time course studied here. Stability of plaques could explain reductions in non-fatal and fatal cardiovascular events associated with increased n-3 PUFA intake. PMID- 12583948 TI - Toe ulcers. PMID- 12583949 TI - MHC typing in variant Creutzfeldt-Jakob disease. AB - Identification of factors that cause susceptibility to, and clinical expression of, variant Creutzfeldt-Jakob disease (vCJD) is essential for future management of the disease. We established MHC genotypes of 76 individuals with vCJD and 131 controls, and analysed MHC phenotypes in relation to age of onset of vCJD and its duration from presentation to death. There were no significant differences between vCJD and control populations in frequencies of any MHC types, nor were there associations between MHC type and age of onset or duration of vCJD disease. Our results do not support the idea of an association between MHC types and either susceptibility to, or expression of, vCJD. PMID- 12583951 TI - Viability and differentiation of autologous skeletal myoblast grafts in ischaemic cardiomyopathy. AB - Autologous skeletal myoblast transplantation might improve postinfarction ventricular function, but graft viability and differentiation (ie, proof of concept) has not been shown. A 72-year-old man had autologous cultured myoblasts from his vastus lateralis injected to an area of transmural inferior myocardial infarction in non-reperfused scar tissue. He showed improvement in symptoms and left-ventricular ejection fraction. When he died 17.5 months after the procedure, the grafted post-infarction scar showed well developed skeletal myotubes with a preserved contractile apparatus. 65% of myotubes expressed the slow myosin isoform and 33% coexpressed the slow and fast isoforms (vs 44% and 0.6%, respectively, in skeletal muscle). Myoblast grafts can survive and show a switch to slow-twitch fibres, which might allow sustained improvement in cardiac function. PMID- 12583950 TI - Serum test for assessment of patients with Bence Jones myeloma. AB - Bence Jones protein in urine (immunoglobulin free-light-chains) is characteristic of light-chain multiple myeloma. We aimed to compare a quantitative immunoassay for serum free-light-chains with urine tests. Of 224 patients with light-chain myeloma tested at entry to clinical trials, all were correctly identified from serum samples. During monitoring of 82 patients, changes in serum and urine free light-chains corresponded, but urine became negative for free-light-chains in 26 patients, whereas it remained abnormal in serum in 73 patients. Serum assays could replace Bence Jones protein urine tests for patients with light-chain multiple myeloma. PMID- 12583952 TI - Humanitarian groups unprepared for Iraq war. PMID- 12583953 TI - US military revamps combat medic training and care. PMID- 12583955 TI - Parasite control can start at school, say African scientists. PMID- 12583957 TI - Russia hunts for funds for ailing health service. PMID- 12583958 TI - Dutch HIV rates on the increase. PMID- 12583959 TI - Uganda tries to learn from Zambia's GM food controversy. PMID- 12583960 TI - Thyroid carcinoma. AB - Thyroid carcinomas are fairly uncommon and include disease types that range from indolent localised papillary carcinomas to the fulminant and lethal anaplastic disease. Several attempts to formulate a consensus about treatment of thyroid carcinoma have resulted in published guidelines for diagnosis and initial disease management. Multimodality treatments are widely recommended, although there is little evidence from prospective trials to support this approach. Surgical resection to achieve local disease control remains the cornerstone of primary treatment for most thyroid cancers, and is often followed by adjuvant radioiodine treatment for papillary and follicular types of disease. Thyroid hormone replacement therapy is used not only to rectify postsurgical hypothyroidism, but also because there is evidence to suggest that high doses that suppress thyroid stimulating hormone prevent disease recurrence in patients with papillary or follicular carcinomas. Treatment for progressive metastatic disease is often of limited benefit, and there is a pressing need for novel approaches in treatment of patients at high risk of disease-related death. In families with inherited thyroid cancer syndromes, early diagnosis and intervention based on genetic testing might prevent poor disease outcomes. Care should be carefully coordinated by members of an experienced multidisciplinary team, and patients should be provided with education about diagnosis, prognosis, and treatment options to allow them to make informed contributions to decisions about their care. PMID- 12583961 TI - Gut flora in health and disease. AB - The human gut is the natural habitat for a large and dynamic bacterial community, but a substantial part of these bacterial populations are still to be described. However, the relevance and effect of resident bacteria on a host's physiology and pathology has been well documented. Major functions of the gut microflora include metabolic activities that result in salvage of energy and absorbable nutrients, important trophic effects on intestinal epithelia and on immune structure and function, and protection of the colonised host against invasion by alien microbes. Gut flora might also be an essential factor in certain pathological disorders, including multisystem organ failure, colon cancer, and inflammatory bowel diseases. Nevertheless, bacteria are also useful in promotion of human health. Probiotics and prebiotics are known to have a role in prevention or treatment of some diseases. PMID- 12583962 TI - Sheehan syndrome. PMID- 12583963 TI - Report of the WHO Commission on Macroeconomics and Health: a summary and critique. PMID- 12583964 TI - 25 years of coronary angioplasty: almost a fairy tale. PMID- 12583965 TI - Keeping scientific advice non-partisan. PMID- 12583966 TI - Heart Protection Study. PMID- 12583967 TI - Heart Protection Study. PMID- 12583968 TI - Heart Protection Study. PMID- 12583970 TI - Heart Protection Study. PMID- 12583972 TI - Cogan's syndrome as an autoimmune disease. PMID- 12583973 TI - HLA in French patients with variant Creutzfeldt-Jakob disease. PMID- 12583974 TI - Low back pain and psychiatric disorders. PMID- 12583975 TI - More on the LIFE study. PMID- 12583976 TI - More on the LIFE study. PMID- 12583979 TI - ERA in International Health: an experiment worthwhile? PMID- 12583980 TI - The hazards of whirlpooling. PMID- 12583982 TI - Samuel Shem. PMID- 12583985 TI - Emergency plan for AIDS relief. PMID- 12583987 TI - Dietary cadmium inhibits spontaneous hepatocarcinogenesis in C3H/HeN mice and hepatitis in A/J mice, but not in C57BL/6 mice. AB - Cadmium is known to be a potent carcinogenic and mutagenic metal. However, we demonstrated that dietary supplementation with 50 ppm cadmium inhibits spontaneous carcinogenesis in C3H/HeN and spontaneous hepatitis in A/J mice. We found that the frequencies of spontaneous hepatocarcinogenesis in C3H/HeN mice and of spontaneous hepatitis in A/J mice fed low-dose cadmium for 54 weeks were significantly lower than those in the respective control groups. A cadmium induced increase in metallothionein production itself and/or metallothionein associated increases in hepatic zinc concentrations may be involved in the observed preventive effects of cadmium. Our results suggest that low doses of cadmium in the diet or environment may play a beneficial role in the prevention of hepatic disease in humans and animals. PMID- 12583988 TI - Transplacental carcinogenicity of inorganic arsenic in the drinking water: induction of hepatic, ovarian, pulmonary, and adrenal tumors in mice. AB - Arsenic is a known human carcinogen, but development of rodent models of inorganic arsenic carcinogenesis has been problematic. Since gestation is often a period of high sensitivity to chemical carcinogenesis, we performed a transplacental carcinogenicity study in mice using inorganic arsenic. Groups (n = 10) of pregnant C3H mice were given drinking water containing sodium arsenite (NaAsO(2)) at 0 (control), 42.5, and 85 ppm arsenite ad libitum from day 8 to 18 of gestation. These doses were well tolerated and body weights of the dams during gestation and of the offspring subsequent to birth were not reduced. Dams were allowed to give birth, and offspring were weaned at 4 weeks and then put into separate gender-based groups (n = 25) according to maternal exposure level. The offspring received no additional arsenic treatment. The study lasted 74 weeks in males and 90 weeks in females. A complete necropsy was performed on all mice and tissues were examined by light microscopy in a blind fashion. In male offspring, there was a marked increase in hepatocellular carcinoma incidence in a dose- related fashion (control, 12%; 42.5 ppm, 38%; 85 ppm, 61%) and in liver tumor multiplicity (tumors per liver; 5.6-fold over control at 85 ppm). In males, there was also a dose-related increase in adrenal tumor incidence and multiplicity. In female offspring, dose-related increases occurred in ovarian tumor incidence (control, 8%; 42.5 ppm, 26%; 85 ppm, 38%) and lung carcinoma incidence (control, 0%; 42.5 ppm, 4%; 85 ppm, 21%). Arsenic exposure also increased the incidence of proliferative lesions of the uterus and oviduct. These results demonstrate that oral inorganic arsenic exposure, as a single agent, can induce tumor formation in rodents and establishes inorganic arsenic as a complete transplacental carcinogen in mice. The development of this rodent model of inorganic arsenic carcinogenesis has important implications in defining the mechanism of action for this common environmental carcinogen. PMID- 12583989 TI - Impact of in vitro gallium arsenide exposure on macrophages. AB - The semiconductor gallium arsenide (GaAs) is classified as an immunotoxicant and a carcinogen. We previously showed that GaAs in vivo induces several phenotypic changes in macrophages located at the exposure site, indicative of an activated state. These physiological alterations may be a primary or secondary consequence of chemical exposure. To discern primary influences, our current study examined the in vitro effects of the chemical on macrophage cell lines and murine peritoneal macrophages. GaAs augmented cathepsins L and B proteolytic activities in all three sources of macrophages. Expression of the two mature isoforms of invariant chain and its cleavage fragment was also significantly increased, indicating that the chemical directly affects macrophages. However, GaAs did not alter the overall cell surface expression of major histocompatibility complex class II molecules on macrophages nor influence their ability to stimulate antigen-specific helper T cell hybridomas to respond to intact antigens that require processing. These findings raise the possibility that the chemical's complete in vivo impact may involve cytokines. Further, GaAs in vitro enhanced steady-state cathepsin L protein, and cathepsins L and B mRNA expression in macrophages, indicating that GaAs may alter gene expression, which may contribute to the chemical's adverse biological effects. PMID- 12583990 TI - Prevention of short-term ultraviolet B radiation-mediated damages by resveratrol in SKH-1 hairless mice. AB - Nonmelanoma skin cancer is the most common cancer among humans and solar UV radiation, particularly its UVB component (290-320 nm), is its major cause. One way to reduce the occurrence of the cancer is via the use of substances (often antioxidants) termed "photochemopreventive agents". Resveratrol (trans-3,4',5 trihydroxystilbene), a phytoalexin found in grapes, nuts, fruits, and red wine, is a potent antioxidant with strong anti-inflammatory and antiproliferative properties. This study was designed to examine whether resveratrol possesses the potential to ameliorate the damages caused by short-term UVB exposure to mouse skin. Single topical application of resveratrol (25 micromol/0.2 ml acetone per mouse) to SKH-1 hairless mice was found to result in significant inhibition of UVB (180 mJ/cm(2))-mediated increase in bifold skin thickness and skin edema. The resveratrol treatment to mouse skin was also found to result in significant inhibition of UVB-mediated induction of cyclooxygenase and ornithine decarboxylase (ODC) enzyme activities and protein expression of ODC, which are well-established markers for tumor promotion. We also observed that resveratrol inhibits UVB-mediated increased level of lipid peroxidation, a marker of oxidative stress. Taken together, our results suggest that resveratrol may afford substantial protection against the damages caused by UVB exposure, and these protective effects may be mediated via its antioxidant properties. PMID- 12583991 TI - Size effect of intratracheally instilled particles on pulmonary inflammation and vascular thrombosis. AB - Particulate air pollution is associated with cardiorespiratory effects and ultrafine particles (UFPs, diameter < 100 nm) are believed to play an important role. We studied the acute (1 h) effect of intratracheally instilled unmodified (60 nm), negatively charged carboxylate-modified (60 nm), or positively charged amine-modified (60 or 400 nm) polystyrene particles on bronchoalveolar lavage (BAL) indices and on peripheral thrombosis in hamster. The latter was assessed by measuring the extent of photochemically induced thrombosis in a femoral vein via transillumination. Unmodified and negative UFPs did not modify thrombosis and BAL indices. Positive UFPs increased thrombosis at 500 microg per animal (+ 341 +/- 96%) and at 50 microg per animal (+ 533 +/- 122%), but not at 5 microg per animal. Neutrophils, lactate dehydrogenase, and histamine were increased in BAL at all these doses but protein concentration was increased only at 500 microg per animal. Positive 400-nm particles (500 microg per animal) did not affect thrombosis, although they led to a neutrophil influx and an increase in BAL proteins and histamine. Using the Platelet Function Analyser (PFA-100), the platelets of hamsters were activated by the in vitro addition of positive UFPs and 400-nm particles to blood. We conclude that intratracheally administered positive ultrafine and 400-nm particles induce pulmonary inflammation within 1 h. Positive UFPs, but not the 400-nm particles enhance thrombosis. Hence, particle induced lung inflammation and thrombogenesis can be partially uncoupled. PMID- 12583992 TI - M40403, a superoxide dismutase mimetic, protects cochlear hair cells from gentamicin, but not cisplatin toxicity. AB - Gentamicin, an aminoglycoside antibiotic, and cisplatin, a platinum-based anticancer drug, are two commonly used clinical drugs with ototoxic side effects. The ototoxicity of gentamicin and cisplatin has been linked to the production of reactive oxygen species (ROS), although the specific ROS pathways have not been identified. One ROS that might play a role in ototoxicity is the superoxide radical, which is enzymatically dismutated to molecular oxygen and hydrogen peroxide by endogenous superoxide dismutase (SOD) enzymes. M40403, a manganese based nonpeptidyl molecule that mimics the activity of SOD, was tested for its ability to protect against gentamicin and cisplatin toxicity in cochlear organotypic cultures from neonatal C57BL/10J mice. Cultures were treated with gentamicin or cisplatin alone or in combination with M40403. M40403 alone had no effect on outer hair cell (OHC) or inner hair cell (IHC) survival at doses of 1, 5, and 10 microM, but a high dose of 30 microM reduced hair cell numbers by approximately 30%. Gentamicin alone and cisplatin alone killed OHCs and IHCs in a dose-dependent manner. The addition of M40403 to gentamicin-treated cultures significantly increased OHC and IHC survival in a dose-dependent manner, whereas M40403 failed to protect hair cells in cisplatin-treated cultures at any dose. The results suggest that the toxicity of gentamicin and cisplatin to cochlear hair cells are mediated by different pathways. Clinically, increased levels of SOD or SOD mimetics might provide significant protection against aminoglycoside ototoxicity. PMID- 12583993 TI - Polychlorinated biphenyls as initiators in liver carcinogenesis: resistant hepatocyte model. AB - A modified Solt-Farber protocol was established to investigate the potential initiating activity of lower chlorinated polychlorinated biphenyl (PCB) congeners in rat liver. Two different studies were conducted in male Fisher 344 rats. PCBs investigated were PCB3, PCB12, PCB38, and PCB77 in study 1 and PCB15, PCB52, PCB77, and the combination of PCB52 and PCB77 in study 2. Rats were subjected to partial hepatectomy followed by a single dose of the suspected initiating agent, diethylnitrosamine, or vehicle. Two weeks later all groups received selection treatment consisting of three daily doses of 2-acetylaminofluorene (2-AAF) and then a single dose of carbon tetrachloride, followed by three additional daily treatments of 2-AAF via gavage. Rats were killed 2 weeks after the last treatment of 2-AAF, and the number and volume of gamma-glutamyltranspeptidase (GGT) positive foci were determined. Among the PCBs tested, PCB3, PCB15, PCB52, and PCB77 significantly increased the number of GGT-positive foci per cm(3) of liver and per liver. Only PCB3 and PCB15 increased the volume fraction of GGT-positive foci. Histopathologic analysis of hematoxylin- and eosin-stained liver sections showed that rats with significantly increased GGT-positive foci also had extensive cellular alteration. This effect was not seen in nonselection groups. We conclude that, under the conditions and time courses of these experiments, several PCBs have initiating activity in male Fischer 344 rats. PMID- 12583995 TI - Development of a gene expression vector for Thermus thermophilus based on the promoter of the respiratory nitrate reductase. AB - A specific expression system for Thermus spp. is described. Plasmid pMKE1 contains replicative origins for Escherichia coli and Thermus spp., a selection gene encoding a thermostable resistance to kanamycin, and a 720 bp DNA region containing the promoter (Pnar), and the regulatory sequences of the respiratory nitrate reductase operon of Thermus thermophilus HB8. Two genes, encoding a thermophilic beta-galactosidase and an alkaline phosphatase were cloned in pMKE1 as cytoplasmic and periplasmic reporters, respectively. The expression of the reporters was specifically induced by the combined action of nitrate and anoxia in facultative anaerobic derivatives of T. thermophilus HB27 to which the gene cluster for nitrate respiration was transferred by conjugation. Overexpressions in the range of approximately 200-fold were obtained for the cytoplasmic reporter, whereas that of the periplasmic reporter was limited to approximately 20-fold, with respect to their intrinsic respective activities. PMID- 12583996 TI - Construction of a novel set of transfer vectors to study vaccinia virus replication and foreign gene expression. AB - Vaccinia virus (VV) is a useful expression vector for many laboratory applications. To date, approximately 60% ( approximately 120) of the VV genes remain uncharacterized. The thought of smallpox being used as a biological weapon has gained attention. In light of this, it is imperative that we continue to study the basic replication of VV, a poxvirus that is closely related to smallpox. A set of plasmid vectors were constructed to generate gene deletions (pZIPPY-NEO/GUS) in VV or for foreign gene expression (pBR-EXPRESS). The vectors contain the Escherichia coli neomycin resistance (neo) and beta-glucuronidase (gusA) genes as selectable markers to facilitate isolation of recombinant viruses. These are the first transfer vectors to use a neo/gusA selection system. We used these vectors to successfully generate a recombinant D9R deletion mutant of VV and to express E. coli lacZ gene. Results indicate that both vectors are highly suited for their designed purpose. PMID- 12583998 TI - Identification of a mouse orthologue of the CED-6 gene of Caenorhabditis elegans. AB - The rapid engulfment of apoptotic cells is a specialized innate immune response used by organisms to remove apoptotic cells. In mammals, several receptors that recognize apoptotic cells have been identified. Previous analysis of the engulfment gene ced-6 in Caenorhabditis elegans (C. elegans) has suggested that CED-6 is an adapter protein that participates in signal transduction pathway that mediates the specific recognition and engulfment of apoptotic cells. Here, we describe our isolation and partial characterization of a mouse cDNA, which is like an orthologue of C. elegans CED-6. PCR screening of mouse cDNA pool with primers designed from the C. elegans CED-6 cDNA sequence resulted in about 300 bp PCR product which was partially sequenced and then screened to a mouse full length cDNA library. Thus in this study we report the identification of a novel C. elegans CED-6-like orthologue in mouse, which has probable apoptotic like function. PMID- 12583997 TI - Construction and evaluation of a plasmid vector for the expression of recombinant lipoproteins in Escherichia coli. AB - Outer membrane lipoproteins are emerging as key targets for protective immunity to many bacterial pathogens. Heterologous expression of lipoproteins in Escherichia coli does not always result in high level expression of acylated recombinant protein. Thus, these proteins do not take up their correct membrane topology and are lacking the immunostimulatory properties endowed by the lipid. To this end, we have designed a lipoprotein expression vector (pDUMP) that results in the production of fusion proteins containing the E. coli major outer membrane lipoprotein (Lpp) signal sequence, lipoprotein signal peptidase recognition site, and the +2 outer membrane sorting signal at their N termini. To test the ability of pDUMP to express lipoproteins from heterologous hosts, the surface lipoprotein PsaA from the Gram-positive organism Streptococcus pneumoniae and the outer membrane lipoproteins MlpA from the Gram-negative Pasteurella multocida and BlpA from the spirochete Brachyspira hyodysenteriae were cloned into both hexahistidine fusion vectors and pDUMP. High level expression of antigenically active protein from both the hexahistidine fusion vectors and pDUMP resulted in abundant bands of the predicted molecular masses when analyzed by SDS PAGE. When grown in the presence of 3[H]palmitic acid, proteins encoded by pDUMP were observed to incorporate palmitic acid whilst the hexahistidine fusion proteins did not. Using mass spectrometry and image analysis we determined the efficiency of lipidation between the three clones to vary from 31.7 to 100%. In addition, lipidated, but not hexahistidine, forms of the proteins were presented on the E. coli surface. PMID- 12583999 TI - Transposon mutagenesis of Mycoplasma gallisepticum. AB - There are few systems available for studying the genetics of the important avian respiratory pathogen, Mycoplasma gallisepticum. These techniques are needed to develop a mechanism to study the molecular pathogenesis of M. gallisepticum. Tn916 has the ability to transpose into the M. gallisepticum genome by both transformation and conjugation. In this study, PEG-mediated transformation was employed for the transfer of Tn916 into M. gallisepticum and create a transposon mutant library. Transformants were obtained at a frequency of approximately 5 x 10(-8) per recipient CFU. A total of 424 MG/Tn916 mutants were constructed and sequence data from the transposon junctions of 71 mutants was obtained and used to identify transposon insertion sites. Insertions were found throughout the genome in nearly all of the major gene categories, making this the first extensive characterization of a transposon mutant library of M. gallisepticum. Transposon stability was also examined, and it was determined that for two mutants the element was stably maintained in vivo in the absence of selective pressure. PMID- 12584000 TI - Characterization of the cryptic plasmid pBGR1 from Bartonella grahamii and construction of a versatile Escherichia coli-Bartonella spp. shuttle cloning vector. AB - We report herein the isolation and molecular characterization of pBGR1, the first native plasmid isolated from the genus Bartonella. Cloning and sequencing revealed a 2725-base pair (bp) cryptic plasmid comprising two open reading frames of considerable length, which were designated rep and mob. The regions containing rep and mob are separated by 140-bp inverted repeat sequences and display a difference in G + C content from one another. A 1435-bp SacI-BclI fragment containing the rep gene is sufficient to mediate replication in the species Bartonella henselae and Bartonella tribocorum, while this replicon does not appear to be functional in Escherichia coli. The Rep protein of 190 amino acids (aa) shares homology to putative replication proteins of cryptic plasmids of Gram negative origin, which form a subgroup of the rolling-circle replication proteins of the pSN2 plasmid superfamily of Gram-positive bacteria. The Mob protein of 333 aa is related to mobilization proteins of several cryptic plasmids and is associated with a conserved recombination site A. The tra functions of RP4 can mobilize pBGR1 derivatives in a mob-dependent manner. Mobilizable pBGR1-based E. coli-Bartonella spp. shuttle vectors were constructed and were shown to be maintained in B. tribocorum during in vivo passage in a rat model in the absence of antibiotic selection. The small size and stability of these shuttle cloning vectors should render them particularly valuable for genetic studies in Bartonella spp. PMID- 12584001 TI - Bacillus subtilis soil isolates: plasmid replicon analysis and construction of a new theta-replicating vector. AB - We have searched for plasmids in a collection of 55 Bacillus subtilis strains isolated from various natural sources of the territory of Belarus. Twenty percent of the strains contained one or two plasmids of either 6-8 or approximately 90 kb. Small plasmids were shown to carry a rolling circle replicon of the pC194 type. Four out of the eight large plasmids contained a related theta replicon that has no homolog in databases as shown by sequence determination. A B. subtilis/Escherichia coli shuttle vector based on this replicon was constructed. It has a low copy number (6 units per chromosome) and is stably inherited in B. subtilis. It might thus be a useful tool for DNA cloning. These data extend previous observations, indicating that most of the small plasmids of B. subtilis replicate as rolling circles and belong to the pC194 family. On the contrary, large plasmids appear to form a large pool of theta-replicating determinants, since three different replicons have already been isolated from them. PMID- 12584002 TI - Insights into the genetic organization of the Corynebacterium diphtheriae erythromycin resistance plasmid pNG2 deduced from its complete nucleotide sequence. AB - The complete nucleotide sequence of the erythromycin resistance plasmid pNG2 from the human pathogen Corynebacterium diphtheriae S601 was determined. The plasmid has a total size of 15,100 bp and contains at least 17 coding regions. Comparative genomics identified conserved motifs within replication initiator proteins of corynebacterial plasmids and a novel nucleotide sequence feature, termed 22-bp box, located downstream of the repA gene. The erythromycin resistance determinant erm(X) is flanked by inverted repeats of the novel insertion sequence IS3504, which may be responsible for a spontaneous deletion of the antibiotic resistance gene region. Furthermore, pNG2 encodes a putative conjugative relaxase, a membrane protein of the natural resistance-associated macrophage protein (Nramp) family and a protein with Nudix hydrolase signature. Expression of the predicted coding regions of pNG2 in Escherichia coli JM109 was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) assays. The detailed annotation of the entire pNG2 sequence provided genetic information regarding its molecular evolution and its role in dissemination of antibiotic resistance genes by horizontal gene transfer. PMID- 12584003 TI - A cosmid vector containing a dominant selectable marker for cloning Chlamydomonas genes by complementation. AB - Cosmid vectors containing a dominate selectable marker (ble) for complementation cloning of genes in Chlamydomonas reinhardtii were created. The usefulness of these vectors, which differ in the orientation of the ble cassette, was demonstrated by transforming C. reinhardtii to phleomycin resistance, by constructing a large library (approximately 5 x 10(5) recombinants) in one of them using DNA from a C. reinhardtii mutant, and by transforming C. reinhardtii with recombinant cosmid clones and pools. PMID- 12584004 TI - Species differences in plasmid carriage in the Enterobacteriaceae. AB - Modern concerns about the spread of antibiotic resistance raise questions about the effect of bacterial species on plasmid evolution and maintenance. We studied 223 Enterobacteriaceae isolated from wild mammals and determined the number of plasmids per isolate, the size of those plasmids, and the distribution of plasmid incompatibility groups N, P, W, FII, and A/C. All of these variables were non randomly distributed with respect to bacterial species, suggesting that host-cell factors constrain the plasmids that a strain will carry. The implication for the evolution of multiple-resistance plasmids in a clinical setting is that although inter-generic plasmid transfer may introduce a novel resistance plasmid into a bacterial genus, it is likely to be modified to suit the requirements of the new host cell. This then further suggests that resistance plasmids will evolve independent lineages within bacterial species although the genes incorporated in them may have come from the same original source. PMID- 12584005 TI - Complete sequencing and analysis of pEN2701, a novel 13-kb plasmid from an endophytic Streptomyces sp. AB - A 12,855 bp cryptic plasmid was isolated from strains of an endophytic Streptomyces sp. over a wide geographical area in South Australia. This plasmid was completely sequenced and 13 putative ORFs were identified. Two of the ORFs may be involved in the regulation of host plant genes. ORF7 exhibited homology to a plant transcriptional regulatory protein and ORF1 was a homolog of a plant protein synthesis initiation factor. The plasmid appears to use a novel transfer mechanism for a Streptomyces plasmid. Pocks were detected during conjugative transfer and kor but not tra homologs could be identified. This structure and the sequence of the putative Kor protein are similar to the pFQ series of plasmids isolated from Frankia, another endophytic actinomycete. PMID- 12584006 TI - Urinary cadmium excretion is correlated with calcaneal bone mass in Japanese women living in an urban area. AB - Nine hundred eight women aged 40-88 years living in a non-Cd-polluted area in Japan were analyzed for urinary cadmium (Cd), N-acetyl-beta-D-glucosaminidase (NAG) activity, beta(2)-microglobulin (B2MG) concentration, and for the stiffness index (STIFF) of calcaneal bone using an ultrasound method. The urinary Cd in the subjects, with a mean and range of 2.87 and 0.25-11.4 microg/g creatinine, respectively, showed a significant correlation with NAG but not with B2MG. STIFF was significantly inversely correlated with urinary Cd, and the association remained significant after adjusting for age, body weight, and menstrual status, suggesting a significant effect of Cd on the bone loss in these subjects without signs of Cd-induced kidney damage. A two-fold increase in urinary Cd was accompanied by a decrease in STIFF corresponding to a 1.7-year rise in age. These results emphasize the need for reassessment of the significance of Cd exposure in the general Japanese population. PMID- 12584007 TI - Trace element intake and status of Italian subjects living in the Gubbio area. AB - Daily intakes of certain trace elements (Pb, Cd, Ni, Hg, and Cr) were assessed using dietary history and weighed record methods and concurrent chemical analysis (CA) of duplicate portions, along with blood levels, in a group (21 M, 23 F) of inhabitants of the Gubbio area (Belvedere, Biscina, Scritto). The evaluation of only intake of trace elements was accomplished in 40 subjects (20 M, 20 F) 1 year later. In both surveys, trace element intakes were generally lower than the potential tolerable weekly intake. However, daily intakes of Pb, Cd, Ni, Hg, and Cr were higher in men compared to women in both surveys (P<0.05). In fact, intakes at the 50th percentile were greater in men by approximately 75% for Pb, 91% for Cd, 20% for Ni, 45% for Hg, and 29% for Cr in the first survey. In the second survey, percentage differences for Pb, Cd, Ni, and Cr were about 12%, 28%, 26%, and 26%, respectively. Blood trace element levels were slightly higher in men in the first survey. Interestingly, no correlation was observed between the intake of trace elements and corresponding blood values. Food basket and total diet, which were computed from the weighed record method for 2 days, showed relevant disagreements with CA of duplicate portion. Moreover, the comparison in content of Pb, Cd, Ni, and Cr obtained by CA of cooked dishes or by calculation using values of raw foods showed significant differences (P<0.05). It was concluded that trace element intake should be assessed by CA of duplicate portion. PMID- 12584008 TI - Deaths related to lead poisoning in the United States, 1979-1998. AB - This study was conducted to describe trends in US lead poisoning-related deaths between 1979 and 1998. The predictive value of relevant ICD-9 codes was also evaluated. Multiple cause-of-death files were searched for records containing relevant ICD-9 codes, and underlying causes and demographic characteristics were assessed. For 1979-1988, death certificates were reviewed; lead source information was abstracted and accuracy of coding was determined. An estimated 200 lead poisoning-related deaths occurred from 1979 to 1998. Most were among males (74%), Blacks (67%), adults of age >/=45 years (76%), and Southerners (70%). The death rate was significantly lower in more recent years. An alcohol related code was a contributing cause for 28% of adults. Only three of nine ICD-9 codes for lead poisoning were highly predictive of lead poisoning-related deaths. In conclusion, lead poisoning-related death rates have dropped dramatically since earlier decades and are continuing to decline. However, the findings imply that moonshine ingestion remains a source of high-dose lead exposure in adults. PMID- 12584009 TI - Bioaccumulation and damaging action of polymetal industrial dust on laboratory mice Mus musculus alba. I. Analysis of Zn, Cu, Pb, and Cd disposition and mathematical model for Zn and Cd bioaccumulations. AB - The concentrations of Zn, Cu, Pb, and Cd in the liver, kidneys, spleen, bones, and carcass of laboratory mice BALB/cy were observed in toxicological experiments. Polymetal industrial dust containing these metals was given to experimental animals at 1% concentration mixed with conventional animal food. Samples for analyses were taken on Days 15, 40, 60, 90, and 120 posttreatment. The experimental data clearly support the established antagonistic interactions among cadmium, zinc, copper, and lead. A mathematical model was proposed to study the main tendencies of heavy metal bioaccumulation under conditions of metal interaction and excessive exposure. The experimental results were assessed on the basis of the model. A rate constant of renal excretion greater than that of hepatic excretion was obtained, which agrees with the observed inversion of cadmium kidney/liver ratio in the conditions of very high exposure. PMID- 12584010 TI - Intrauterine bisphenol A exposure leads to stimulatory effects on Sertoli cell number in rats. AB - Using the optical disector for quantifying cell numbers, we investigated whether oral treatment of rats on days 6-21 of gestation with the weakly estrogenic bisphenol A (BPA, 0.1 or 50 mg/kg) or the highly estrogenic ethinyl estradiol (EE, 0.02 mg/kg) alters testicular histology, in those offspring 9-12 month of age. Since production of male germ cells depends on Sertoli cell number, possible changes in that parameter were investigated using unbiased stereology. Spermatogenesis was qualitatively normal in all groups. BPA increases Sertoli cell number per organ but not when expressed as per gram testis. EE did not affect cell number per organ but did affect numbers on a per gram testis basis due to a lowered testis weight. In contrast to the lowering of Sertoli cell numbers that might have been expected according to the estrogen hypothesis, intrauterine administration of these xenoestrogens in fact resulted in minor increases in Sertoli cell numbers and had no qualitative effect on spermatogenesis. PMID- 12584011 TI - Effects of Aroclor 1242 and different fish-based diets on vitamins A1 (retinol) and A2 (3,4-didehydroretinol), and their fatty acyl esters in mink plasma. AB - The effects of a 21-week exposure to Aroclor 1242 (1mg per day in feed) on plasma concentrations of vitamins A(1) (retinol) and A(2) (3,4-didehydroretinol) and their principal fatty acyl esters (A(1)-16:0, A(2)-16:0 (palmitates), A(1)-18:1n 9; A(2)-18:1n-9 (oleates), and A(1)-18:0; A(2)-18:0 (stearates)) were studied in young female mink (Mustela vison) fed a diet based on freshwater smelt. These vitamin levels were also examined in mink fed diets containing Baltic herring or fatty marine fish. In the Aroclor-exposed smelt-fed mink, the plasma concentrations of A(1) and A(2) esters were significantly lower than the levels in controls fed the uncontaminated smelt diet. In addition, the A(2) esters reacted more sensitively to the polychlorinated biphenyls than did A(1) esters. In contrast, in the plasma of the exposed mink the level of alcoholic A(1) was normal, and transport of thyroxine (T(4)) and nonspecific lipoprotein transport of major lipids were not impaired. Despite the large dietary supply of vitamin A(2) and high levels of plasma A(2) esters, the mink fed freshwater smelt had only trace amounts of alcoholic vitamin A(2) in their plasma. The concentrations of A(1) and A(2) esters in the plasma of all the mink studied correlated with the hepatic total concentrations of the vitamins. Thus, in carnivores that have nonspecific lipoprotein transport of vitamin A esters, determination of plasma levels of the esterified vitamins may be a useful nondestructive way to estimate stores of the vitamin A analogs in the body and to assess the organochlorine induced decrease in the vitamin stores. PMID- 12584012 TI - Effects of chromium exposure from a cement factory. AB - The aim of the study was to determine the chromium concentrations of soil and plant specimens taken from a rural area exposed to cement factory emissions and also to determine the blood concentrations and sensitivity conditions observed in humans residing in this rural area. The study was carried out in Cukurhisar, a town in Eskisehir-Turkey, between May 2000 and March 2001. Besides the 108 soil (36 for control) and plant specimens, which were taken from eight different directions from the cement factory, blood samples of the individuals residing in this area were taken from 258 subjects (258 for control) following a physical examination, and patch tests were also applied. The chromium concentrations of the soil and plant specimens taken from different places in different directions of the factory were higher than in the control areas. The physical examination of subjects did not reveal results different from those of the control group except for the diagnosis of contact dermatitis. The analyses of venous blood samples showed that chromium concentrations were found to be within the reference values given for both groups, but higher in the subjects (P<0.001). According to the results of patch tests, sensitivity to chromium was found to be more frequent for the subject group than the control group (P<0.05). According to these results, clinical tools revealed no toxic effects for the subjects, except contact dermatitis. However, sensitivity to patch tests showed that this subject group has been affected compared to the control group and that this effect increased with age. PMID- 12584013 TI - Mobility, bioavailability, and toxic effects of cadmium in soil samples. AB - Total concentration is not a reliable indicator of metal mobility or bioavailability in soils. The physicochemical form determines the behavior of metals in soils and hence the toxicity toward terrestrial biota. The main objectives of this study were the application and comparison of three approaches for the evaluation of cadmium behavior in soil samples. The mobility and bioavailability of cadmium in five selected soil samples were evaluated using equilibrium speciation (Windermere humic aqueous model (WHAM)), extraction procedures (Milli-Q water, DMSO, and DTPA), and a number of bioassays (Microtox, growth inhibition test, contact toxicity test, and respiration). The mobility, represented by the water-extractable fraction, corresponded well with the amount of cadmium in the soil solution, calculated using the WHAM (r(2)=0.96, P<0.001). The results of the ecotoxicological evaluation, which represent the bioavailable fraction of cadmium, correlated well with DTPA extractability and also with the concentration of free cadmium ion, which is recognized as the most bioavailable metal form. The results of the WHAM as well as the results of extraction experiments showed a strong binding of cadmium to organic matter and a weak sorption of cadmium to clay minerals. PMID- 12584014 TI - Potential for rhizofiltration of uranium using hairy root cultures of Brassica juncea and Chenopodium amaranticolor. AB - Hairy root cultures of Brassica juncea and Chenopodium amaranticolor were developed by genetic transformation using Agrobacterium rhizogenes. The stable, transformed root systems demonstrated a high growth rate of 1.5-3.0 g/g dry weight/day in Murashige and Skoog medium. In the present study, hairy root system was used for removal of uranium from the solution of concentration up to 5,000 microM. The results indicated that the hairy roots could remove uranium from the aqueous solution within a short period of incubation. B. juncea could take up 20 23% of uranium from the solution containing up to 5,000 microM, when calculated on g/g dry weight basis. C. amaranticolor showed a slow and steady trend in taking up uranium, with 13% uptake from the solution of 5,000 microM concentration. Root growth was not affected up to 500 microM of uranium nitrate over a period of 10 days. PMID- 12584015 TI - Seeing patterns in the noise. AB - How do observers detect the presence of objects or features in visual images? Stochastic stimuli (for example, white noise) have become popular choices for providing a linear characterization of early sensory mechanisms. A recent paper by Neri and Heeger takes this type of methodology a step further, and succeeds in isolating and characterizing non-linear mechanisms responsible for the detection and identification of a specific visual target. PMID- 12584016 TI - Imagined movements that leak out. AB - In a case study that fundamentally alters our understanding of motor imagery, Schwoebel et al. report a patient who unintentionally carries out imagined movements. Furthermore, his 'imagery' movements are more accurate than his intended movements, which suggests that the inhibitory signal that normally prevents us from acting out our motor imagery can be selectively blocked. Removing this inhibition allows us to observe motor imagery 'in action', and reveals that motor imagery and motor planning for execution are not identical. PMID- 12584017 TI - Do we all look alike to computers? AB - People are better able to recognize faces of their own race than those of other races. One explanation is that this 'other-race effect' is caused by lifelong experience in which faces from some races are more common than other races. A recent article by Furl et al. tests experience-based accounts of the other-race effect by giving computer algorithms of face recognition training on preponderantly Caucasian faces. The only algorithms that reliably produced other race effects were those that created face representations distorted to emphasize features that individuated faces. PMID- 12584018 TI - An ABSURDIST model vindicates a venerable theory. AB - Can relations between concepts in one system be sufficient on their own for locating matching concepts in another system? This question in concept semantics has been contentious. Now, contrary to one important theory, the answer appears to be yes. Robert Goldstone's and Brian Rogosky's new neural-net program, ABSURDIST, has demonstrated what purely theoretical considerations suggested could not be accomplished, showing once again the power of experimental computer modeling. PMID- 12584019 TI - Human spatial representation derived from a honeybee compass. PMID- 12584020 TI - How seriously should we take Minimalist syntax? PMID- 12584021 TI - Linguistics and empirical evidence. Reply to Edelman and Christiansen. PMID- 12584022 TI - Capturing underlying differentiation in the human language system. PMID- 12584023 TI - Differentiation and integration in human language. Reply to Marslen-Wilson and Tyler. PMID- 12584024 TI - The mind's best trick: how we experience conscious will. AB - We often consciously will our own actions. This experience is so profound that it tempts us to believe that our actions are caused by consciousness. It could also be a trick, however - the mind's way of estimating its own apparent authorship by drawing causal inferences about relationships between thoughts and actions. Cognitive, social, and neuropsychological studies of apparent mental causation suggest that experiences of conscious will frequently depart from actual causal processes and so might not reflect direct perceptions of conscious thought causing action. PMID- 12584025 TI - Moving towards solutions to some enduring controversies in visual search. AB - How do we find a target item in a visual world filled with distractors? A quarter of a century ago, in her influential 'Feature Integration Theory (FIT)', Treisman proposed a two-stage solution to the problem of visual search: a preattentive stage that could process a limited number of basic features in parallel and an attentive stage that could perform more complex acts of recognition, one object at a time. The theory posed a series of problems. What is the nature of that preattentive stage? How do serial and parallel processes interact? How does a search unfold over time? Recent work has shed new light on these issues. PMID- 12584026 TI - Functional imaging of 'theory of mind' AB - Our ability to explain and predict other people's behaviour by attributing to them independent mental states, such as beliefs and desires, is known as having a 'theory of mind'. Interest in this very human ability has engendered a growing body of evidence concerning its evolution and development and the biological basis of the mechanisms underpinning it. Functional imaging has played a key role in seeking to isolate brain regions specific to this ability. Three areas are consistently activated in association with theory of mind. These are the anterior paracingulate cortex, the superior temporal sulci and the temporal poles bilaterally. This review discusses the functional significance of each of these areas within a social cognitive network. PMID- 12584027 TI - Grounding conceptual knowledge in modality-specific systems. AB - The human conceptual system contains knowledge that supports all cognitive activities, including perception, memory, language and thought. According to most current theories, states in modality-specific systems for perception, action and emotion do not represent knowledge - rather, redescriptions of these states in amodal representational languages do. Increasingly, however, researchers report that re-enactments of states in modality-specific systems underlie conceptual processing. In behavioral experiments, perceptual and motor variables consistently produce effects in conceptual tasks. In brain imaging experiments, conceptual processing consistently activates modality-specific brain areas. Theoretical research shows how modality-specific re-enactments could produce basic conceptual functions, such as the type-token distinction, categorical inference, productivity, propositions and abstract concepts. Together these empirical results and theoretical analyses implicate modality-specific systems in the representation and use of conceptual knowledge. PMID- 12584028 TI - The psycholinguistics of metaphor. AB - Can lawyers be sharks, can jobs literally be jails, and can dogs fly across lawns? Such metaphors create novel categories that enable us to characterize the topic of interest. These novel metaphorical categories are special in that they are based on outstanding exemplars of those categories, and they borrow the exemplar's name for use as the category names. Thus 'shark' can be taken as a metaphor for any vicious and predatory being. Contemporary research reveals how people can create and understand such metaphors in ordinary conversation, and suggests that we understand metaphorical meanings as quickly and automatically as we understand literal meanings. PMID- 12584029 TI - Interleukin-6 and interleukin-11 support human osteoclast formation by a RANKL independent mechanism. AB - Interleukin-6 (IL-6) and interleukin-11 (IL-11) are known to influence osteoclast formation and bone resorption. In order to determine whether IL-6 and IL-11 could independently support human osteoclast formation, these factors were added to cultures of human peripheral blood mononuclear cells of the monocyte (CD14(+)) fraction in the presence of macrophage colony-stimulating factor (M-CSF). Under these conditions, IL-6 and IL-11 induced the formation of multinucleated cells which were positive for TRAP, VNR, and calcitonin receptor and capable of lacunar resorption. Osteoclastogenesis induced by IL-6 and IL-11 was inhibited by the addition of an anti-gp130 antibody but not by osteoprotegerin. These results indicate that IL-6 and IL-11, which are thought to play a role in several osteolytic bone disorders, are directly capable of inducing osteoclast formation by a RANKL-independent mechanism. PMID- 12584030 TI - Does serum cholesterol contribute to vertebral bone loss in postmenopausal women? AB - Recent in vitro and animal studies suggest that cholesterol and its metabolites inhibit the functional activity of osteoblasts and thereby induce reduced bone mineralization. However, scant information is available on the clinical implication of these findings with special regard to postmenopausal bone loss. Therefore, the aim of the present study was to investigate cross-sectional and longitudinal associations between serum cholesterol, bone mineral density (BMD), and bone turnover in 340 postmenopausal women aged 50-75 years (mean 59 years), who were followed for 8.3 +/- 1.1 years. BMD in the lumbar spine, distal forearm, and total hip was measured by dual energy X-ray absorptiometry. Other study variables were physical measures, serum cholesterol, serum markers of bone turnover, and self-reported information on various risk factors for osteoporosis. At baseline, serum cholesterol showed significant negative correlation with BMD at the lumbar spine (r = -0.21, P < 0.0001) and distal forearm (r = -0.14, P = 0.013), but not at the hip. No associations of serum cholesterol with serum osteocalcin (r = 0.054, P = 0.317) and CTX (r = -0.027, P = 0.623) were, however, noted. After adjustment for age and BMI, the negative correlation remained significant at the lumbar spine (r = -0.16, P = 0.004), but not at the distal forearm (r = -0.018, P = 0.738). At the end of the 8-year follow-up, the correlation between serum cholesterol and spine BMD was not observed. Those with the largest increases in serum cholesterol, however, showed the greatest decreases in spine BMD independently of the changes in BMI (r = -0.16, P = 0.004). The correlation between the changes in serum cholesterol and the simultaneous changes in osteocalcin (r = 0.081, P = 0.140) and CTX (r = 0.042, P = 0.441) were statistically insignificant. Thus, our results suggest that the weak associations between spine BMD and serum cholesterol can be explained by the fact that both variables are simultaneously affected by estrogen deficiency rather than by a direct influence of serum cholesterol on osteoblast function. PMID- 12584031 TI - Changes in serum HDL and LDL cholesterol in patients with Paget's bone disease treated with pamidronate. AB - Amino bisphosphonates represent one of the most important advances in the management of Paget's and other metabolic bone diseases. Although their mechanism of action has not yet been completely clarified, they seem to inhibit the mevalonate pathway and so they could interfere with cholesterol synthesis. The present study aimed to evaluate cholesterol and lipoprotein serum levels in patients with Paget's bone disease treated with intravenous pamidronate. The study included 20 consecutive patients (mean age, 67.6 +/- 11.0 years) with Paget's bone disease for at least 1 year, who needed intravenous amino bisphosphonate treatment; 12 patients with inactive Paget's bone disease served as controls. The patients with active Paget's bone disease underwent three cycles (every 3 months) of treatment with 60 mg of intravenous pamidronate. Controls were given a saline infusion following the same administration schedule. In all subjects total alkaline phosphatase (total ALP), bone alkaline phosphatase (bone ALP), total cholesterol (TC), tryglycerides (TG), and high- and low-density lipoprotein cholesterol (HDL-C and LDL-C, respectively) were measured before infusions (pamidronate or saline) at baseline and at 3-month intervals up to 9 months. In the control group no significant changes were observed through the study period for any of the biochemical parameters. In the pamidronate-treated patients, both bone ALP and total ALP significantly fell at the end of the study. In patients with active treatment, at the end of the study period HDL-C significantly (P < 0.05) increased by 10.3%, whereas LDL-C significantly (P < 0.05) decreased by 5.5%. In these patients TC showed a negative trend without reaching statistical significance, whereas the HDL-C/LDL-C ratio rose 16.2% above the basal value and TC/HDL-C decreased by 12.5%. In conclusion, pamidronate given intravenously seems to be able to induce a prolonged shifting in circulating cholesterol from the LDL-C to the HDL-C from associated with a weak decrease in total cholesterol, thus producing a possible improvement in the atherosclerotic risk index. PMID- 12584032 TI - An immunoassay for type I collagen alpha 1 helicoidal peptide 620-633, a new marker of bone resorption in osteoporosis. AB - Type I collagen fragments are the most sensitive markers of bone resorption in osteoporosis. Currently, all available type I collagen-related bone resorption markers detect in serum and/or urine fragments arising from the telopeptide region of the molecule. Our aim was to evaluate the technical and clinical performances of a new assay detecting in urine a degradation fragment originating from the helical part of type I collagen and consisting of the 620-633 sequence of the alpha1 chain. Urinary helical peptide was measured with a new ELISA (Metra Helical peptide, Quidel Corporation). Results were compared with those of urinary C-terminal cross-linking type I collagen of type I collagen (CTX), an established bone resorption marker. We measured urinary helical peptide levels in 89 healthy women (age 31-89 years) and in 59 postmenopausal women involved in two randomized studies of the efficacy of alendronate (10 mg/day; n = 20) and transdermal 17beta estradiol (50 microg/day; n = 39). The within-run and between-run CVs were < or = 7.3 and 8.7%, respectively. In 59 healthy women, urinary helical peptide levels highly correlated with those of urinary CTX (r = 0.78, P < 0.0001). The long-term intraindividual variability assessed over 6 months in 18 untreated postmenopausal women was 24%. Compared to 24 premenopausal women, urinary helical peptide was 42% (P < 0.0001) higher in 65 postmenopausal women (mean age, 60 years), an increase comparable to that of urinary CTX (+ 47%, P < 0.0001). Urinary helical peptide levels decreased by 72% (P < 0.0001) after 3 months of alendronate treatment and by 59% (P < 0.0001) after 6 months of transdermal estrogen therapy. These changes were of similar magnitude to those of urinary CTX (-69 and -62%, respectively; NS compared with changes in helical peptide). In women treated with transdermal 17beta estradiol, the percentage of change of urinary helical peptide and urinary CTX at 6 months significantly correlated with the change in spinal bone mineral density after 2 years (r = -0.58, P = 0.002, and r = -0.52, P = 0.006, for urinary helical peptide and CTX, respectively). This new assay for type I collagen helical peptide has demonstrated adequate analytical performance and was highly correlated with urinary CTX, an established type I collagen C telopeptide bone resorption marker. The test was a sensitive indicator of the antiresorptive effects of bisphosphonate and estrogens in postmenopausal women. This new bone resorption marker should be useful for the clinical investigation of patients with osteoporosis. PMID- 12584033 TI - Coordinated development of embryonic long bone on chorioallantoic membrane in ovo prevents perichondrium-derived suppressive signals against cartilage growth. AB - Perichondrium has been shown to elicit signals to suppress differentiation and proliferation of chondrocytes during endochondral bone formation based on in vitro organ culture. However, these in vitro organ cultures did not allow the growth of bone collar, and thus the effect of perichondrium in a normal environment where development of adjacent embryonic tissues, including bone collar, is taking place has not yet been fully understood. Therefore, we examined the effect of perichondrium on cartilage development using chicken long bone organ cultures on chorioallantoic membrane in ovo, which supported bone collar development. In contrast to previous observations in in vitro organ cultures, in ovo organ cultures prevented overgrowth of epiphyseal cartilage due to the removal of perichondrium. This prevention was associated with the suppression of aggrecan gene expression in the absence of perichondrium in ovo. These results indicated that the perichondrium-derived activity that was observed in vitro to suppress cartilage development could be counterbalanced in ovo, where culture conditions are closer to those in in vivo. TUNEL assay indicated enhanced apoptosis in the presence of perichondrium in vitro, and removal of the perichondrium suppressed apoptosis. No major apoptosis was observed in ovo regardless of the presence or the absence of perichondrium. Thus, chondrogenesis in long bone could be coordinately regulated through modulation of apoptosis by perichondrium and adjacent embryonic tissues, including bone collar, as revealed in in ovo assay. PMID- 12584034 TI - Quality assessment of the cortical bone of the human mandible. AB - This study is the first to investigate simultaneously both the regional and the directional variations in mechanical properties and computed tomography (CT) numbers of the fresh mandible bone and to explore the correlation between the two sets of data with a view to provide a noninvasive method for determining the bone quality for designers of dental implants. Using a three-point bending test the regional variation of Young's modulus of bone in the human mandible was determined from five fresh specimens from donors representative of patients in need of dental implants in that some of the mandibles were fully dentulate, some partly so, and some fully edentulate. While a pattern of the modulus distribution was evident in the mandibles we tested, these mandibles did have, as a result of their respective states, their own peculiarities. We determined also the directional variations of the modulus for the mandible, which are due to the anisotropic nature of bone. The modulus values obtained in our study were much lower than those listed in the only other published study, which was conducted on dry mandibles. These differences in the modulus data from the two studies were attributed partly to the differences in condition of the bone and differences in the dimensions of specimens tested in the two studies. Because the values of the modulus obtained in this study are lower than those previously published, they would be safer as a basis for implant designs. A weak correlation was found between the modulus values and the CT number of the mandible. This would not be sufficient for accurate predictions of the bone properties from CT scans. The development of a noninvasive method for determining the bone quality in various patients thus remains a challenge for researchers. PMID- 12584035 TI - Intermittent intravenous administration of the bisphosphonate ibandronate prevents bone loss and maintains bone strength and quality in ovariectomized cynomolgus monkeys. AB - Using a clinically relevant regimen, this study investigated the effects of treatment with ibandronate, a highly potent nitrogen-containing bisphosphonate, on bone loss, biochemical markers of bone turnover, densitometry, histomorphometry, biomechanical properties, and bone concentration in aged ovariectomized monkeys. Sixty-six female cynomolgus monkeys, aged 9 years and older, were ovariectomized (OVX) or sham operated. Intravenous (iv) bolus injections of ibandronate at 10, 30, or 150 microg/kg or placebo were administered at 30-day intervals (corresponding to intervals of 3 months in humans), starting at OVX, for 16 months. OVX significantly decreased bone mass at the lumbar spine, proximal femur, femoral neck, and radius and increased bone turnover in a time-dependent manner, as assessed by dual energy X-ray absorptiometry, peripheral quantitative computed tomography, or histomorphometry. Ibandronate iv bolus injections administered at 30 microg/kg every 30 days prevented osteopenia induced by estrogen depletion. OVX-induced increases in bone turnover (as determined by activation frequency, bone formation rate, and biochemical markers of bone turnover, including urinary N-telopeptide and deoxypyridinoline excretion and serum values for osteocalcin and bone-specific alkaline phosphatase) were suppressed on treatment, and bone mass, architecture, and strength were preserved at clinically relevant sites. Treatment with high dose (150 microg/kg/dose) iv bolus injections of ibandronate further increased bone mass and improved bone strength at both the spine and femoral neck, without adversely affecting bone quality. In contrast, treatment with a 10 microg/kg/dose only partially prevented the OVX-induced effects. These data support the potential for the long-term administration of ibandronate by intermittent iv bolus injections in humans to prevent osteoporosis and improve bone quality at clinically relevant sites. PMID- 12584036 TI - Bone density changes in Paget's disease 2 years after iv pamidronate: profound, sustained increases in pagetic bone with severity-related loss in forearm nonpagetic cortical bone. AB - Bone mineral density (BMD) was measured at three sites (forearm, spine, and hip) using dual X-ray and single-photon absorptiometry in 68 patients with Paget's disease before and after treatment with iv pamidronate. Patients were treated according to the severity of their disease; the mild category (Group I, hydroxyproline excretion (Hyp(E)) <5.0 micromol/L GF) received 120 mg, the moderate category (Group II, Hyp(E) 5.0-9.99 micromol/GF) 180 mg, and the severe category (Group III, > or = 10.0 micromol/GF) 240 mg. Group I was followed for 1 year, and both Groups II and III for 2 years. At the lumbar spine in pagetic bone there were no differences between groups in early responses, with a profound increase 6 months after treatment 20.5 +/- 2.0% above baseline values to 1.403 +/ 0.063 g/cm(2) (mean +/- SEM)(P < 0.001). This increase in BMD was sustained to 2 years (1.355 +/- 0.078 g/cm(2), P < 0.001) and was 15.0 +/- 2.2% above baseline values. The pagetic total hip BMD increased after treatment in all groups, with a mean rise of 10.4 +/- 1.4% at 1 year to 1.505 +/- 0.083 g/cm(2) (P < 0.01). At the pagetic femoral neck the response was similar, with a peak significant rise at 1 year of 10.7 +/- 1.7% to 1.403 +/- 0.097 g/cm(2) (P < 0.01). In nonpagetic spinal bone there were no differences between the group responses, with a combined mean increase of 4.3 +/- 0.7% at 1 year to 0.999 +/- 0.027 g/cm(2) (P < 0.01). In both Groups II and III the increase in BMD was significantly higher than baseline values at 1 and 2 years (P < 0.01). In the nonpagetic total hip BMD remained unchanged over the 2-year period and likewise, there were no significant changes from baseline at the nonpagetic femoral neck site. In the nonpagetic forearm we found a significant loss in BMD at the ultradistal (mainly trabecular), midregion (80% cortical), and proximal shaft (95% cortical) sites in Group III, persisting to 2 years at the latter two sites. The increase in bone density in pagetic bone, persisting at least 2 years, provides a new modality of assessment of the response of pagetic bone to treatment and suggests a mechanism for the reduction in fracture risk in such bone after effective bisphosphonate treatment. Severity-dependent nonpagetic forearm bone loss, persisting to 2 years at cortical sites, suggests a potential drug-induced fracture risk at the forearm and possibly elsewhere in the absence of appropriate preventive cotreatment. PMID- 12584037 TI - Modeling elite male athletes' peripheral bone mass, assessed using regional dual x-ray absorptiometry. AB - There is still considerable debate as to whether bone mineral content (BMC) increases in proportion to the projected bone area, A(p), or an estimate of the skeletal bone volume, (A(p))(3/2), being assessed. The results from this study suggest that the bone mass acquisition of elite athletes' arms and legs increases in proportion to the projected bone area, A(p), having simultaneously controlled/removed the effect of the confounding variables of body mass and body fat. Although this supports the use of the traditional bone mineral density ratio (BMD=BMC/A(p)), it also highlights the dangers of overlooking the effect of known confounding variables. Ignoring the effect of such confounding variables, athletic groups whose activities involve upper body strength (rugby, rock climbing, kayaking, weight lifting) had the highest arm BMD, while runners were observed to have the lowest arm BMD (lower than that of the controls). Similarly, leg BMD was highest in rugby players, whose activities included both running and strength training. However, the rugby players were also observed to have the greatest body mass. When the important determinants of body mass, body fat, as well as projected bone area, A(p), were incorporated as covariates into a proportional allometric ANCOVA model for BMC, different conclusions were obtained. The introduction of these covariates had the effect of reducing the sporting differences on adjusted arm BMC, although the "sport" by "side" interaction still identified racket players as the only group with a greater dominant arm BMC (P < 0.05). In contrast, sporting differences in adjusted leg BMC remained highly significant, but with a rearranged hierarchy. The runners replaced the rugby players as having the greatest adjusted leg BMC. The results confirm the benefits of activity on peripheral bone mass as being site-specific but reinforce the dangers of making generalizations about the relative benefits of different exercises ignoring the effects of known confounding variables, such as body size, body composition, and age. PMID- 12584038 TI - Low-intensity, high-frequency vibration appears to prevent the decrease in strength of the femur and tibia associated with ovariectomy of adult rats. AB - The effect of low-intensity, high-frequency vibration on bone mass, bone strength, and skeletal muscle mass was studied in an adult ovariectomized (OVX) rat model. One-year-old female rats were allocated randomly to the following groups: start control, sham OVX, OVX without vibration, OVX with vibration at 17 Hz (0.5g), OVX with vibration at 30 Hz (1.5g), OVX with vibration at 45 Hz (3.0g). Vibrations were given 30 min/day for 90 days. During vibration each group of rats was placed in a box on top of the vibration motor. The amplitude of the vibration motor was 1.0 mm. The animals were labeled with calcein at day 63 and with tetracycline at day 84. The tibia middiaphysis was studied by mechanical testing and dynamic histomorphometry and the femur distal metaphysis by mechanical compression. OVX without vibration increased the periosteal bone formation rate and increased the medullary cross-sectional area, i.e., increased the endocortical resorption and outward anteromedial and lateral drifts of cortical bone at the tibia middiaphysis. OVX also resulted in a reduced maximum bending stress of the tibia diaphysis and a reduced compressive stress of the femur distal metaphysis. Vibration at the highest intensity, i.e., 45 Hz, of OVX rats induced a further increase in periosteal bone formation rate and inhibited the endocortical resorption seen in OVX rats. Furthermore, vibration at 45 Hz inhibited the decline in maximum bending stress and compressive stress induced by OVX. Neither OVX nor OVX with vibration influenced skeletal muscle mass. In conclusion, the results support the idea of a possible beneficial effect of passive physical loading on the preservation of bone in OVX animals. PMID- 12584039 TI - Independent predictors of all osteoporosis-related fractures in healthy postmenopausal women: the OFELY study. AB - Several epidemiological studies have identified clinical factors that predict the risk of hip fractures in elderly women independently of the level of bone mineral density (BMD), such as low body weight, history of fractures, and clinical risk factors for falls. Their relevance in predicting all fragility fractures in all postmenopausal women, including younger ones, is unknown. The objective of this study was to identify independent predictors of all osteoporosis-related fractures in healthy postmenopausal women. We prospectively followed for 5.3 +/- 1.1 years a cohort of 672 healthy postmenopausal women (mean age 59.1 +/- 9.8 years). Information on social and professional conditions, demographic data, current and past medical history, fracture history, medication use, alcohol consumption, caffeine consumption, daily calcium intake, cigarette smoking, family history of fracture, and past and recent physical activity was obtained. Anthropometric and total hip bone mineral density measurements were made. Incident falls and fractures were ascertained every year. We observed 81 osteoporotic fractures (annual incidence, 21 per 1000 women/year). The final model consisted of seven independent predictors of incident osteoporotic fractures: age > or = 65 years, odds ratio estimate (OR), 1.90 [95% confidence interval (CI) 1.04-3.46], past falls, OR, 1.76 (CI 1.00-3.09), total hip bone mineral density (BMD) < or = 0.736 g/cm(2), OR, 3.15 (CI 1.75-5.66), left grip strength < or = 0.60 bar, OR, 2.05 (CI 1.15-3.64), maternal history of fracture, OR, 1.77 (CI 1.01-3.09), low physical activity, OR, 2.08 (CI 1.17-3.69), and personal history of fragility fracture, OR, 3.33 (CI 1.75-5.66). In contrast, body weight, weight loss, height loss, smoking, neuromuscular coordination assessed by three tests, and hormone replacement therapy were not independent predictors of all fragility fractures after adjustment for all variables. We found that some--but not all--previously reported clinical risk factors for skeletal fragility predicted all fragility fractures independently of BMD in healthy postmenopausal women, although they differed somewhat from those predicting specifically hip fractures in elderly women. These risk factors appear to reflect quality of bone structure (previous fragility fracture), lifestyle habits (physical activity), muscle function and health status (grip strength), heredity (maternal history of fracture), falls, and aging. Measurements of these variables should be included in the clinical assessment of the risk of osteoporotic fractures in postmenopausal women. PMID- 12584040 TI - Increased femoral neck cancellous bone and connectivity in coxarthrosis (hip osteoarthritis). AB - Patients with coxarthrosis (cOA) have a reduced incidence of intracapsular femoral neck fracture, suggesting that cOA offers protection. The distribution of bone in the femoral neck was compared in cases of coxarthrosis and postmortem controls to assess the possibility that disease-associated changes might contribute to reduced fragility. Whole cross-section femoral neck biopsies were obtained from 17 patients with cOA and 22 age- and sex-matched cadaveric controls. Densitometry was performed using peripheral quantitated computed tomography (pQCT) and histomorphometry on 10-microm plastic-embedded sections. Cortical bone mass was not different between cases and controls (P > 0.23), but cancellous bone mass was increased by 75% in cOA (P = 0.014) and histomorphometric cancellous bone area by 71% (P < 0.0001). This was principally the result of an increase of apparent density (mass/vol) of cancellous bone (+45%, P = 0.001). Whereas cortical porosity was increased in the cases (P < 0.0001), trabecular width was also increased overall in the cases by 52% (P < 0.001), as was cancellous connectivity measured by strut analysis (P < 0.01). Where osteophytic bone was present (n = 9) there was a positive relationship between the amount of osteophyte and the percentage of cancellous area (P < 0.05). Since cancellous bone buttresses and stiffens the cortex so reducing the risk of buckling, the increased cancellous bone mass and connectivity seen in cases of cOA probably explain, at least in part, the ability of patients with cOA to resist intracapsular fracture of the femoral neck during a fall. PMID- 12584041 TI - Serum osteoprotegerin is a major determinant of bone density development and prevalent vertebral fracture status following cardiac transplantation. AB - Osteoprotegerin (OPG) is an antiresorptive cytokine and a key regulator of osteoclastogenesis and activity. Since OPG is downregulated by glucocorticoids and cyclosporine A in vitro we examined whether immunosuppressive therapy would play a role in the development of transplantation osteoporosis. We enrolled 57 cardiac transplant recipients (median time since transplantation, 3.2 years (1.1 11.5 years)) in this cross-sectional study. Standardized spinal X-rays as well as hip bone density measurements were performed in all patients. Serum OPG was determined using a commercially available ELISA. Vertebral fractures were present in 56% of the patients. Bone densities of all femoral neck subregions were correlated to serum OPG concentrations (r values between 0.40 and 0.48, all P < 0.005). Multiple regression analysis revealed OPG levels to be independently correlated to femoral neck Z scores (r = 0.49, P = 0.002). After adjustment for age, BMI, neck Z score, renal function, and months since transplantation, serum OPG was the only significant predictor of prevalent vertebral fractures (P = 0.001). In a separate 6-month prospective study of 14 heart transplant recipients receiving calcium and vitamin D serum OPG levels fell by 41% (P = 0.0004) after 3 months and 47% (P = 0.0001) after 6 months following cardiac transplantation. Bone loss at the lumbar spine and femoral neck after 6 months was correlated to the decrease in serum OPG at 6 months (r = 0.82, P < 0.0001, and r = 0.60, P = 0.02, respectively) as well as 3 months after cardiac transplantation (r = 0.65, P = 0.01, and r = 0.69, P = 0.006, respectively). Serum OPG alone accounted for 67% of the variance of lumbar spine bone density changes over the first 6 months posttransplantation. We conclude that serum OPG levels decline consistently in all patients following initiation of immunosuppressive therapy and are independently correlated with changes in bone density. We hypothesize that OPG plays a major role in the development of transplantation osteoporosis. PMID- 12584043 TI - MHC multimers: expanding the clinical toolkit. PMID- 12584044 TI - Adoptive T cell therapy--immune monitoring and MHC multimers. PMID- 12584045 TI - Induction of immunity to tumor-associated antigens following dendritic cell vaccination of cancer patients. PMID- 12584046 TI - HLA tetramer-based artificial antigen-presenting cells for stimulation of CD4+ T cells. AB - Current evidence suggests that immunotherapy for cancer or infectious diseases will require the activation of CD4(+) T cells in addition to the activation of cytotoxic CD8(+) T cells. To complement and overcome some of the limitations of dendritic-cell-based vaccines and ex vivo expansion of human T cells, we sought to engineer artificial antigen-presenting cells (aAPCs) for the stimulation of antigen-specific human CD4(+) T cells. We have designed a variety of aAPCs using magnetic beads as a scaffold on which to coat HLA-peptide tetrameric complexes along with costimulatory molecules such as anti-CD28. Here, we tested various forms of conjugation of the tetramers onto beads, characterized the relative concentration of antigen available on the surface of the beads, and evaluated the ability of different types of beads to promote activation of antigen-specific CD4(+) T cells. We find that an indirect coating of HLA-peptide tetramers on beads via an anti-Class II antibody provides specific stimulation of antigen specific CD4(+) T cells. PMID- 12584047 TI - Challenges in staining T cells using HLA class II tetramers. PMID- 12584048 TI - Diagnostic potential of tetramer-based monitoring of cytomegalovirus-specific CD8+ T lymphocytes in allogeneic stem cell transplantation. AB - Cytomegalovirus (CMV) infection remains a significant problem in allogeneic stem cell transplant (SCT) recipients despite the availability of effective antiviral drugs. This problem concerns patients which are unable to mount an effective T lymphocyte response against CMV. Therefore, the development and use of tetramer technology to enumerate CMV-specific T cells will be valuable to identify these patients as early as possible. Here, we review clinical studies in which CMV specific CD8(+) T cells have been monitored in allogeneic SCT recipients using tetramers in the context of similar studies that are based on functional assays of CMV-specific T cells. The results thus far warrant the further development of tetramer technology as a diagnostic tool to monitor CMV-specific T cells in SCT recipients and other groups of immunocompromised patients threatened by CMV. PMID- 12584049 TI - Chemokine receptor genotype and response to interleukin-2 therapy in HIV-1 infected individuals. AB - Interleukin-2 therapy is an immune-based treatment for HIV-1-infected individuals with declining CD4(+) T cell counts. Intravenous IL-2 produces an elevation of circulating CD4(+) T cells, but with a varying degree of effectiveness in individual patients. IL-2 is also known to increase the expression of chemokine receptors, coreceptors for HIV-1. Allelic variation in chemokine receptor genes can markedly affect the course of HIV disease; consequently, we analyzed CCR5 and CCR2B genotypes among a cohort of HIV-1-infected individuals that received IL-2 therapy. DNA was extracted from treated individuals and genotyping was performed using PCR followed by allele-specific detection or cleavage of the amplified product. Samples from 47 trial participants (25 CIV-IL-2 group; 22 placebo group) were analyzed for CCR5 and CCR2B genotype. We report that CCR5 Delta 32 heterozygous individuals had a greater CD4(+) T cell response to continuous intravenous IL-2 (CIV-IL-2) treatment than those homozygous for the wild-type allele (median = 427 vs 237 cells/mm(3); P = 0.03). This study highlights the importance of interactions between IL-2 and CCR5; at the clinical level, it argues for assessment of chemokine receptor genotype in IL-2 and perhaps other immune-based therapy trials. PMID- 12584050 TI - Defective activity of ERK-1 and ERK-2 mitogen-activated protein kinases in peripheral blood T lymphocytes from patients with systemic lupus erythematosus: potential role of altered coupling of Ras guanine nucleotide exchange factor hSos to adapter protein Grb2 in lupus T cells. AB - The integrity of the Ras/Raf/mitogen-activated protein kinase (MAPK) cascade is critical for maintenance of T cell tolerance, a process that fails in patients with systemic lupus erythematosus (SLE). In this study we have examined the activity of mitogen-activated protein kinases ERK-1 and ERK-2 in resting and TCR activated peripheral blood T lymphocytes from patients with SLE. We also examined the binding of Ras guanine nucleotide exchange factor, human Son of Sevenless (hSos), to cytosolic adapter protein growth factor receptor-bound protein 2. T cells from lupus patients showed diminished catalytic activity and TCR-driven dual phosphorylation of ERK-1 and ERK-2 upon stimulation through the TCR/CD3 receptor, a defect that may be related to altered translocation of hSos to the Ras/Raf membrane complex and diminished nuclear translocation of trans-acting factor AP-1. Defective MAPK activity triggered by TCR/ CD3 activation may alter the coordination of signals needed for normal interleukin-2 production and maintenance of tolerance in lupus T cells. PMID- 12584051 TI - Up-regulated expression of MICA and proinflammatory cytokines in skin biopsies from patients with seborrhoeic dermatitis. AB - Seborrhoeic dermatitis is a disease of unknown etiopathogenesis that affects 5% of the population. In this study, we investigated expression of mRNA for IL-1 alpha, IL-6, IL-4, IFN-gamma, and the stress-inducible MICA molecule in skin biopsies from 12 patients with moderate to severe seborrhoeic dermatitis and 2 healthy volunteers by RT-PCR and hybridization with specific probes. Eight patients expressed INF-gamma, 2 expressed IL-6, 8 expressed IL-1 alpha, and 2 expressed IL-4 (1 with moderate disease). Eight patients expressed inflammatory cytokines (IL-1 alpha, IL-6, and/or IFN-gamma) in healthy skin. Higher cytokine mRNA in damaged vs healthy skin was also observed, suggesting the existence of an inflammation that predisposes healthy skin to develop overt disease. Up-regulated expression of MICA mRNA was observed in 8 patients. Although the pathogenesis of seborrhoeic dermatitis remains to be elucidated, expression of cytotoxicity activating ligands (MICA), recruitment of NK cells, and a local pro-inflammatory microenvironment may facilitate the development of tissue injury. PMID- 12584052 TI - C5 is required for CD49d expression on neutrophils and VCAM expression on vascular endothelial cells following mesenteric ischemia/reperfusion. AB - Complement activation is critical in the development of local mucosal damage and inflammation as well as of remote organ injury after mesenteric ischemia/reperfusion. To further define the role of C5 activation in local and remote tissue injury, C5 deficient (C5(-/-)) and wild-type control (C5(+/+)) mice treated with an anti-C5 mAb were subjected to sham or ischemia followed by up to 4 h of reperfusion. The development of local (intestinal) and remote (lung) injury was associated with the expression of CD49d on the surface of circulating blood neutrophils and with VCAM on the endothelial cells of intestinal and lung vessels. Because CD49d heterodimerizes with integrin beta1 on the surface of neutrophils and can bind VCAM on the endothelium, we propose that complement activation causes organ damage by upregulating molecules that lead to inappropriate homing of neutrophils. PMID- 12584053 TI - Monocyte response to Th1 stimulation and effector function toward human mesangial cells are not impaired in patients with lupus nephritis. AB - Monocytes/macrophages activated by Th1 stimulation such as interferon-gamma (IFN gamma) and CD40 ligand (CD40L) infiltrate the kidney and play a critical role in the progression of lupus nephritis (LN). We examined the monocyte response to Th1 stimulation and their effector function toward activating renal resident cells in patients with LN. Following stimulation with IFN-gamma granulocyte macrophage colony stimulating factor (GM-CSF)/recombinant CD40L the production of tumor necrosis factor-alpha and IL-12 p70 by PBMC was significantly higher in LN patients. In coculture experiments employing activated monocytes and human mesangial cells, there was a trend toward higher monocyte chemoattractant protein 1 production by lupus monocytes compared to normal controls. Basal expression of CD40, ICAM-1, and STAT-1 was significantly higher in monocytes from LN patients, suggesting ongoing activation. Monocyte response to IFN-gamma, as accessed by intercellular adhesion molecule-1 upregulation and phosphorylation of STAT-1, was comparable between the two groups. Thus, in contrast to earlier reports, Th1 dependent monocyte activation is not impaired. In this disease activated monocytes appear to be fully capable of inducing renal injury. PMID- 12584054 TI - The logic of social sharing: an evolutionary game analysis of adaptive norm development. AB - Although norms can potentially serve useful constructs to understand human minds, being fundamentally social in evolutionary as well as cultural senses, there are as yet no useful psychological theories of adaptive norm development. This article provides an illustrative model about how a norm emerges in a society. We focus on the "communal-sharing norm" in primordial societies, a norm designating uncertain resources as common properties to be shared with other members. Based on anthropological findings, we develop a theory about how the communal-sharing norm emerges and is maintained. Then, using evolutionary computer simulations, we test several hypotheses about the conditions under which the norm will dominate social resource sharing. We further test behavioral implications of the norm, demonstrating that uncertainty involved in resource acquisition is a key factor that triggers the psychology of sharing even in highly industrialized societies. Finally, we discuss the importance of norm construct for analyzing the dynamic relation between minds and society. PMID- 12584055 TI - The existential function of close relationships: introducing death into the science of love. AB - Originally, terror management theory proposed two psychological mechanisms in dealing with the terror of death awareness-cultural worldview validation and self esteem enhancement. In this article, we would like to promote the idea of close relationships as an additional death-anxiety buffering mechanism and review a growing body of empirical data that support this contention. Based on a comprehensive analysis of the sociocultural and personal functions of close relationships, we formulate two basic hypotheses that have received empirical support in a series of experimental studies. First, death reminders heighten the motivation to form and maintain close relationships. Second, the maintenance of close relationships provides a symbolic shield against the terror of death, whereas the breaking of close relationships results in an upsurge of death awareness. In addition, we present empirical evidence supporting the possibility that close relationships function as a related yet separate mechanism from the self-esteem and cultural worldview defenses. PMID- 12584056 TI - Implicit theories of relationships: orientations toward evaluation and cultivation. AB - Implicit theories of relationships (ITRs) influence goals, motivations, attributions, and behavior in romantic relationships. We developed a model of ITRs that draws from social cognition, motivation, and achievement literatures, and derived conceptual parallels and hypotheses with regard to relationships. It is proposed that ITRs reflect the belief component of a larger system of motivations and goals that can influence the degree to which people are oriented toward the evaluation and cultivation of relationships. Research on ITRs is reviewed with regard to how they moderate well-documented associations between relationship perceptions and outcomes. Differences between ITRs and implicit theories in other domains are also discussed. PMID- 12584057 TI - Relational discrepancy theory: the implications of self discrepancy theory for dyadic relationships and for the emergence of social structure. AB - Relationship formation and maintenance is guided by more than similarity between partners' actual selves. Expanding the domain of self-discrepancy theory (SDT; Higgins, 1987), we propose that a type of discrepancy previously not considered discrepancies between self-aspects of relational partners-is central to relationship cognition, including perceptions of intimacy and trust, and to the emergence of roles within relationships. Our argument relates both to unconstrained environments, where individuals freely choose partners, and constrained environments (e.g., workplaces) with relationships imposed. We argue that SDT's prediction of emotional consequences from discrepancies permits a motivational account of why individuals might form and maintain relationships in terms of hierarchies and roles PMID- 12584058 TI - Artificial a theoretical model of triggered displaced aggression. AB - A tit-for-tat matching rule (Axelrod, 1984) describes much interpersonal behavior. Yet, in daily life a retaliatory aggressive response to a trivially mild provocation often inappropriately exceeds that expected from the matching rule. The concept of triggered displaced aggression can explain these exceptions to the matching principle. Building from the Cognitive Neoassociationistic model of aggressive behavior (Berkowitz, 1989, 1990, 1993), we developed a theoretical framework of social and personality factors that moderate and mediate the disjunctively escalated retaliation that can result from triggered displaced aggression. Major explanatory factors in our analysis of such effects are as follows: (a) aspects of the Time 1 provocation and the immediate situation in which it occurred; (b) characteristics of initial provocations and personality factors of the actor that produce the ruminative thought that will temporally extend the effects of a Time 1 provocation, allowing them to interact with a delayed Time 2 minor triggering event; and (c) actions and attributes of the target of displaced aggression that augment these effects. PMID- 12584062 TI - 2002 Bruno Koppler Distinguised Contribution Award. Paradigms of personality assessment: an interpersonal odyssey. PMID- 12584063 TI - Bell object relations inventory for adolescents and children: reliability, validity, and factorial invariance. AB - The Bell Object Relations Inventory (BORI; Bell,1995) is a self-report instrument that measures deficits in object relations ego functioning. It has demonstrated clinical and research utility in adult populations. This article reports the development of a version of the BORI for children ages 11 to 17, including studies of reliability, validity, and factorial invariance. Data of 705 children from public schools and 110 children from clinics and residential treatments were used. Of the 45 original BORI true/false items, 3 were dropped as inappropriate and most others were rewritten for easier reading. An additional 16 items were created to capture unique features of adolescent experiences in relationships. Items were tested in focus groups and revised accordingly. Eight items were dropped because of low communalities, so that 50 items were included in the final factor analysis. Assessments using self-report items from Behavioral Assessment Scale for Children (BASC; Reynolds & Camphaus, 1992) and the Personality Inventory for Youth (PIY) were also obtained to test concurrent validity. Oblique rotation yielded 5 factors. Four were very similar to the 4 from the adult version and were named accordingly: Alienation, Insecure Attachment, Egocentricity, and Social Incompetence. The fifth scale was comprised mostly of new items and was called Positive Attachment. Scales showed excellent factorial invariance and good internal consistency. Scales generally had very low intercorrelations reflecting their relative independence. Although differences were found for gender and race, the effect sizes were small. Support for construct validity came from moderate correlations with concurrent BASC and PIA scores, analyses of variance showing greater deficits in object relations in pathological subgroups compared with normals, and a trend analysis showed that Alienation scores followed a lawful relationship with increasing severity of psychopathology. These initial findings support the psychometric integrity of the instrument and its construct validity. Results indicate continuity of constructs between the adult and adolescent versions. The Positive Attachment subscale may prove a valuable addition for clinical and research applications. PMID- 12584064 TI - Self-report bias and underreporting of depression on the BDI-II. AB - One problem in identifying and treating depression is underreporting of symptoms by individuals. Previous research suggests that there may be systematic sex differences in self-report bias,with men tending to minimize their depressive symptoms more than women. This study used an experimental design with a sample of 238 community members to test whether disguising the purpose of the Beck Depression Inventory-II would significantly reduce self-report bias, especially in men. We found a main effect of condition such that both men and women reported significantly more core depressive symptoms in the covert condition, suggesting that surveys of community samples may underestimate the prevalence of depression. PMID- 12584065 TI - The psychometric characteristics of the Hamilton Depression Inventory. AB - In this study, the psychometric properties of the Hamilton Depression Inventory (HDI; Reynolds & Kobak, 1995a) were examined in a sample of 249 undergraduate participants. The HDI exhibited high internal consistency and support for its construct validity was demonstrated by the HDI's patterns of correlations with other measures of depression, anxiety, and depression-relevant cognition. Factor analyses of the full (23-item) and 17-item versions of the HDI each yielded 4 factors, which accounted for 49% and 53% of the variance in participants' responses, respectively. The utility of the HDI's use of multiple-weighted subitems was also assessed by comparing a less complicated scoring system to the standard scoring format. The standard HDI added significantly to the prediction of criterion indexes after controlling for the variance accounted for by the "simplified" HDI. Moreover, the operating characteristics of the standard HDI outperformed the simplified HDI in the prediction of the Beck Depression Inventory-II (Beck, Steer, & Brown, 1996) classification. The results provide strong support for the HDI as a reliable and valid instrument for the assessment of depressive severity PMID- 12584066 TI - Rorschach measures of aggressive drive derivatives: a college student sample. AB - This study investigated measures of aggressive drive derivatives on the Rorschach and Personality Assessment Inventory with 70 college students. As predicted, (a) self-reported physical aggression potential was related to Rorschach measures of identification with the aggressor and aggressive impulses, (b) suicidal ideation with impulsivity was related to a Rorschach measure of aggressive impulses turned toward the self, and (c) the borderline features scale was related to a Rorschach pathological object relations measure. Hierarchical regression analyses showed that a Rorschach measure of emotional impulsivity added unique variance to these Rorschach aggression variables in predicting self-reported physical aggression potential, suicidal ideation with impulsivity, and borderline features. Finally, caution is advised in applying our findings when there is motivation to suppress aggressive responses PMID- 12584067 TI - The use of the Ego Impairment Index across the schizophrenia spectrum. AB - The goal of this study was to assess perceptual and thought disturbance, as indexed by the Ego Impairment Index (EII; Perry & Viglione, 1991), a Rorschach derived measure, across the schizophrenia spectrum. We hypothesized that there would be an increase in EII scores (indicating increased disturbance) across the spectrum from nonpatients to severely disturbed, hospitalized schizophrenia patients. Normal comparison participants (n = 66), students with elevated scores on either the Perceptual Aberration/Magical Ideation or the Physical Anhedonia Scales (n = 24), first-degree relatives of schizophrenia patients (n = 36), participants diagnosed with Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) schizotypal personality disorder (n = 36), outpatient schizophrenia participants (n = 33), and hospitalized schizophrenia patients (n = 56) were studied. As hypothesized, we found increased EII scores in all of the schizophrenia spectrum groups when compared against normal comparison participants. Furthermore, the EII was significantly different between the schizophrenia patients and the other schizophrenia spectrum groups. These findings support the use of the EII as a sensitive measure of perceptual and thought disturbance across the schizophrenia spectrum that yields specific information regarding the type of thinking problems that occur within schizophrenia spectrum subgroups. PMID- 12584068 TI - Beck Depression Inventory-II items associated with self-reported symptoms of ADHD in adult psychiatric outpatients. AB - The Beck Depression Inventory-II (BDI-II; Beck, Steer, & Brown, 1996) and the Conners' Adult ADHD Rating Scale-Self-Report: Screening Version (CAARS-S:SV; Conners, Erhardt, & Sparrow, 1999) were administered to 371 (64%) female and 204 (36%) male adult (> 18 years old) outpatients who were diagnosed with various psychiatric disorders to determine whether any of the 21 items or subsets of items in the BDI-II were related to symptoms of attention deficits and hyperactivity as measured by the CAARS-S:SV DSM-IV Total ADHD Symptoms scale (attention-deficit/hyperactivity disorder [ADHD] Symptoms). Stepwise multiple regression analyses found that the BDI-II Concentration Difficulty explained 30% of the variance in these total scores. Ratings > 1 for the BDI-II Concentration Difficulty item were discussed as being useful for ruling out possible symptoms of ADHD. PMID- 12584069 TI - Construct validity of the relationship profile test: a self-report measure of dependency-detachment. AB - This study evaluated the construct validity of the Relationship Profile Test (RPT; Bornstein & Languirand, 2003), a 30-item self-report measure of dependency detachment that yields three subscale scores: (a) destructive overdependence, (b) dysfunctional detachment, and (c) healthy dependency. Scores on the RPT subscales generally showed the expected patterns of intercorrelations and gender differences, and comparison of RPT scores with scores on other tests supported the convergent and discriminant validity of each RPT subscale. Results of internal and retest reliability analyses were generally supportive as well, and suggested that the three RPT subscales assess aspects of the traits they purport to measure PMID- 12584070 TI - Measuring epistemic curiosity and its diversive and specific components. AB - A questionnaire constructed to assess epistemic curiosity (EC) and perceptual curiosity (PC) curiosity was administered to 739 undergraduates (546 women, 193 men) ranging in age from 18 to 65. The study participants also responded to the trait anxiety, anger, depression, and curiosity scales of the State-Trait Personality Inventory (STPI; Spielberger et al., 1979) and selected subscales of the Sensation Seeking (SSS; Zuckerman, Kolin, Price, & Zoob, 1964) and Novelty Experiencing (NES; Pearson, 1970) scales. Factor analyses of the curiosity items with oblique rotation identified EC and PC factors with clear simple structure. Subsequent analyses of the EC items provided the basis for developing an EC scale, with Diversive and Specific Curiosity subscales. Moderately high correlations of the EC scale and subscales with other measures of curiosity provided strong evidence of convergent validity. Divergent validity was demonstrated by minimal correlations with trait anxiety and the sensation-seeking measures, and essentially zero correlations with the STPI trait anger and depression scales. Male participants had significantly higher scores on the EC scale and the NES External Cognition subscale (effect sizes of r =.16 and.21, respectively), indicating that they were more interested than female participants in solving problems and discovering how things work. Male participants also scored significantly higher than female participants on the SSS Thrill-and Adventure and NES External Sensation subscales (r =.14 and.22, respectively), suggesting that they were more likely to engage in sensation-seeking activities. PMID- 12584071 TI - Psychological measures as predictors of military training performance. AB - The predictive validity of 7 ability tests, the Big Five, and the Rorschach method administered to 71 male applicants at the Naval Special Forces (NSF) of Norway was evaluated based on pass/fail results in training. The findings showed: (a) small correlations between the ability tests, the Big Five scales, and the success criterion; (b) Rorschach variables measuring stress tolerance, reality testing, cognition, and social adjustment correlated significantly (r =.25 to.48) with pass/fail results in training, and (c) logistic regression analysis revealed that 3 of the Rorschach variables accumulated incrementally in the prediction of training completion when entered after the ability tests and the Big Five scales, thus supporting the merit of using Rorschach variables for predicting NSF training performance PMID- 12584072 TI - Starting at the beginning: an introduction to coefficient alpha and internal consistency. AB - Cronbach's a is the most widely used index of the reliability of a scale. However, its use and interpretation can be subject to a number of errors. This article discusses the historical development of a from other indexes of internal consistency (split-half reliability and Kuder-Richardson 20) and discusses four myths associated with a: (a) that it is a fixed property of the scale, (b) that it measures only the internal consistency of the scale, (c) that higher values are always preferred over lower ones, and (d) that it is restricted to the range of 0 to 1. It provides some recommendations for acceptable values of a in different situations. PMID- 12584073 TI - Neuropsychological performance of patients following mold exposure. AB - This study investigated the effects of mold exposure (ME) on human cognition by analyzing neuropsychological data from patients who were exposed to mold in their homes or workplaces. Compared to normative data, ME patients were impaired (<10th percentile) on a number of cognitive measures, with the most consistent deficits in visuospatial learning, visuospatial memory, verbal learning, and psychomotor speed. We also examined emotional functioning and found that a number of ME patients showed evidence of both Axis I and Axis II pathology. Interestingly, there was a significant correlation among patients' scores on the Beck Depression Inventory-Second Edition and the number of neuropsychological tests falling within the impaired range. Given the limited understanding of ME and its effect on the human central nervous system, we provide a working model that attempts to capture the complex interactions of impaired cognition, psychosocial stressors, poor physical health, and emotional functioning in patients following ME. KEYWORDS: mold exposure, mild traumatic brain injury, toxic exposure PMID- 12584074 TI - Longitudinal Wisconsin card sorting performance in schizophrenia patients in rehabilitation. AB - Recent investigations into the course of cognitive functioning in schizophrenia have not been able to determine whether cognitive functioning declines, remains stable, or improves over time. This report analyzes Wisconsin Card Sorting Test (WCST) and Wechsler Adult Intelligence Test-Revised Digit Symbol Substitution Test (WAIS-R DSST) performance over time to determine if there is deterioration, improvement, or stability of performance. Data for 46 participants, diagnosed with schizophrenia or schizoaffective disorder that were in a stable phase of illness and actively involved in rehabilitation, were gathered from assessments made at 2 time points on average 4.3 years apart. Results indicated that there was stability in most WCST variables; however, there was significant improvement on WCST perseverative error and on the DSST. Sixty-five percent of the sample improved on 1 or both of the test variables. Furthermore, cognitive test changes were unrelated to type of psychotropic medication and only modestly related to symptom variables. Results indicate that in a stable schizophrenia sample, WCST performance is generally stable over the 3- to 7-year time period, but there are some cognitive variables that may improve even without direct intervention. KEYWORDS: schizophrenia, cognition, executive function, rehabilitation, stability PMID- 12584075 TI - Computer-related anxiety: examining the impact of technology-specific affect on the performance of a computerized neuropsychological assessment measure. AB - This study was conducted to examine the effect of impairment status and computer specific anxiety on the performance of a computerized neuropsychological assessment measure. Computer related anxiety was measured using a standardized self-report measure tapping anxiety specific to computers and technology. Outcome on this measure was compared with error scores and response timing variables on a computerized version of the Category Test (CT) in both normal individuals and individuals with neurological, psychiatric, or substance abuse histories. Multivariate analysis results, controlling for psychomotor performance, revealed significant main effects for group status and computer-related anxiety. CT performance was significantly related to the level of computer-related anxiety, in that high anxiety resulted in higher CT error scores and longer response times, and the negative impact of computer-related anxiety on computerized neuropsychological assessment performance was stronger in individuals with impairment histories. Our results suggest that as computer-related anxiety increases, performance on computer administered neuropsychological assessment measures tends to decrease. Key words: computers, anxiety, computer-based task performance, clinical neuropsychology, Category Test PMID- 12584077 TI - Global affective memory for faces in patients with temporal lobe epilepsy. AB - Eighteen (9 left, 9 right) epileptic patients with unilateral temporal lobe epilepsy (TLE) and 20 control participants were evaluated. Participants were instructed to make global affective judgments of likeability in a face memory task or to make neutral piecemeal judgments of nose size during the encoding phase of the task. A main effect of better memory after using a global affective versus a neutral piecemeal encoding strategy was found. Patients with right sided abnormality performed more poorly in the global affective encoding condition than patients with left-sided abnormality or than the control participants. No differences were seen among groups for the neutral piecemeal condition. Further, unlike the left TLE patients and the control participants, the right TLE patients did not show better performance when using the global affective encoding strategy over the neutral piecemeal strategy. This is discussed in relation to the literature suggesting an important role for the right hemisphere in global affective processing and memory, as well as the literature indicating an important role for the amygdala in affective memory. PMID- 12584076 TI - The 12-item Buschke memory test: appropriate for use across levels of impairment. AB - Monitoring cognitive functions as older adults move from independent to assisted living is of utmost importance with respect to care planning. To this end, we examined the utility of a 12- item, free and cued recall, selective reminding, memory task for assessing persons across levels of functioning. Using cross sectional data from the Canadian Study of Health and Aging, it was observed that the 12-item Buschke memory test was well tolerated by participants regardless of their level of impairment. Further, various measures from the memory task differentiated among participants with different levels of impairment: Individuals living in the community performed better than those in institutions; individuals with no or mild functional impairment performed better than individuals with moderate or severe functional impairments; individuals with mild dementia performed better than those with moderate to severe dementia. Normative data are provided for this easily administered and well-tolerated memory measure. Key words: cognitive assessment, memory test, dementia, levels of impairment PMID- 12584078 TI - Raven's progressive matrices performance in adults with traumatic brain injury. AB - Raven's Progressive Matrices (RPM), a widely used test of reasoning, is sensitive to aging, but it has not proven to be helpful in the assessment of acquired focal or lateralized brain damage. Clinical experience suggests that the test is insensitive to traumatic brain injury (TBI), but the data are difficult to interpret because of rapid inflation of norms over time (the Flynn effect). In examining data from 64 adult patients with TBI who were administered the Standard RPM between 1981 and 1989, we used previous and subsequent norms conjointly to adjust for the Flynn effect. Anterograde and retrograde adjustment of norms led to highly convergent results. After adjustment for the Flynn effect, RPM performance was comparable to Wechsler IQ, significantly below estimated premorbid IQ, and nearly 2 SD above performance on 2 TBI-sensitive neuropsychological tests. We conclude that RPM performance is neither more nor less sensitive than Wechsler IQ to the consequences of TBI in the adult, but erroneous conclusions are likely to be reached if the Flynn effect is not taken into account. PMID- 12584079 TI - Neuropsychological test performance of Swedish multiple chemical sensitivity patients--an exploratory study. AB - To address the hypothesis of brain dysfunction as a component of the multiple chemical sensitivity (MCS) syndrome, a neuropsychological battery comprising 8 tests was given to 17 Swedish MCS patients and 34 demographically matched controls. Across the 6 tests used as indicators of brain impairment, comprising a total of 17 test variables, the MCS group performed poorer only in a complex reaction time test (mean reaction time; p = 0.002; t test). Correction for self ratings of mental distress and trait psychasthenia did not eliminate the deviation in the reaction time test. Because the results on most tests were within normal limits, brain impairment was not evidenced. However, the similar minor deviations in neurobehavioral tests observed in several studies of MCS patients indicate the need for a study on a larger sample of MCS cases. PMID- 12584080 TI - Subjective experience after stroke. AB - To assess subjective experience after stroke, 214 patients completed the 63-item European Brain Injury Questionnaire. The same questionnaire was also completed by a close relative of each patient and by 214 control participants matched for gender and age. A principal component analysis showed 3 factors: depressive mood, cognitive difficulties, and difficulties in social interactions. In self assessment, whereas the first 2 factors were scored higher by patients than by controls, the level of difficulty in social interactions did not differ between these 2 groups. In hetero-assessment, relatives scored patients' difficulties higher than patients did, especially for items suggesting the impact of stroke on themselves. Psychological difficulties of stroke patients and the consequences on their relatives need to be taken into account in rehabilitation programs. PMID- 12584081 TI - Emotional perception in unilateral stroke patients: recovery, test stability, and interchannel relationships. AB - Despite an ever-increasing literature on language and cognitive recovery after brain injury, there are relatively few investigations about the recovery of emotional processing. The main purpose of this study was to provide a preliminary evaluation of recovery of emotional perception across 3 communication channels in unilateral stroke patients. In addition, instrument stability and interrelationships among the channels were examined. Tasks assessing facial, prosodic, and lexical emotional identification from the New York Emotion Battery (Borod, Welkowitz, & Obler, 1992) were administered to right-brain-damaged (RBD), left-brain-damaged (LBD), and normal control (NC) participants. Emotional, as well as nonemotional control, tasks were examined at 2 times, with a median interval of 25 months. Findings revealed some evidence of recovery on emotional perception tasks. Participant group differences correlations were high for NCs and LBDs but low for RBDs. Significant relationships were more frequent for the facial versus prosodic channel than for the lexical versus the 2 nonverbal channels, suggesting that facial and prosodic perception may subserve a general emotional processor. PMID- 12584082 TI - Does high scatter affect the predictive validity of WAIS-III IQs? AB - We tested the assumption that high amounts of intersubtest scatter on the Wechsler Adult Intelligence Scale-Third Edition (WAIS-III) subtest profiles compromise predictive validity of the IQs for predicting Wechsler Memory Scale Third Edition (WMS-III) indexes. Data from a sample of 80 male Veteran's Administration medical center patients were analyzed, half with high intersubtest scatter and half with low scatter. The 2 groups were matched on Full Scale IQ. Correlations of WAIS-III Full Scale IQ with WMS-III indexes were not significantly different between the 2 groups. Further, the regression equations for predicting WMS-III indexes did not depend on the amount of scatter. The results suggest that, when differences in IQ are controlled, the validity of WAIS III scores in predicting memory performance does not depend on the amount of intersubtest scatter. Further research is needed with samples from different populations using a variety of criterion variables. PMID- 12584083 TI - Analysis of substitution test performance using eye movement and video data. AB - Substitution tests continue to enjoy widespread popularity in applied neuropsychology. Though highly sensitive, these tests lack specificity, and substitution test decrements are often reported in terms that bear little relation to cognitive or other psychological theory. Objectives were to quantify, using eye movement/video analysis, subitem performance elements for the Wechsler Adult Intelligence Scale-Revised (WAIS-R) Digit Symbol Test (DST); to assess the relative contribution of each of these to standard WAIS-R DST performance in a group; and to assess whether wearing the eye-movement measuring equipment interfered with test performance. PMID- 12584084 TI - Factor structure of MicroCog in a clinical sample. AB - Two factor solutions adequately describe the MicroCog performance data of a clinical sample of substance abusers. Information Processing Speed and Information Processing Accuracy factors seem to explain the Short Form and Standard Form scores of a clinical sample as they do for the normative sample. Clinicians need to be aware that the factor scores do not allow for unambiguous interpretation, but may be more meaningful than independent domain scores. PMID- 12584085 TI - Tests of executive function predict instrumental activities of daily living in community-dwelling older individuals. AB - The purpose of this study was to examine the utility of specific tests of executive functioning for predicting instrumental activities of daily living (IADLs) in community-dwelling older individuals (n = 30). In addition to tests of frontal and executive functioning, performance-based and caregiver-rated evaluations of IADLs were obtained. Results indicated that different tests of executive functioning were associated with outcomes on performance-based versus caregiver-rated assessments of IADLs. Specifically, the Trail Making Test (Part B) made a significant and unique contribution to the prediction of performance based IADLs. In contrast, verbal fluency performance and Trail Making Test performance made significant independent contributions to the prediction of IADLs as reported by a caregiver. These findings suggest that different aspects of frontal and executive functions may be related to IADLs, depending on the type of assessment instrument used. Key words: activities of daily living, executive function, older adults PMID- 12584086 TI - The medical school's faculty is its most important asset. PMID- 12584088 TI - Developing a new faculty practice plan with a model for funds flow between the hospital and the plan. AB - The majority of academic health centers are experiencing significant difficulties balancing their research, teaching, and clinical missions while maintaining adequate financial performance. One of the major areas under intense scrutiny is the specific financial relationship between the hospital and the full-time faculty. A realignment of the funds flow between these two entities is becoming essential to the future viability and ultimate survival of many health systems. The authors describe a model that evolved as part of the integration of the faculty practice plans of their institutions when they merged into a single health system, and that provides a framework that specifically addresses these issues of funds flow. The model includes (1) a strong partnership between the hospital and the full-time faculty; (2) a governance model of chairmen, faculty, and administration; (3) flexibility for the department chairs to set salaries and make significant financial decisions relative to their departmental performances; (4) a specific formula for funds flow for graduate medical education dollars from the hospital to the clinical departments; (5) local front-end charge capture and back-end central collections; and (6) clear and consistent definitions of revenue and expense items for both partners. PMID- 12584089 TI - Are search committees really searching? AB - Academic chair searches are admittedly a labor-intensive process, but they are made more difficult and often lead to less-than-optimal outcomes because search committees spend their time "advertising," "looking," but not truly searching for academic chairs. At the onset, certain "realities" must be acknowledged, including (1) understanding that unless your organization is renowned in the specialty for which you are conducting the search, candidates won't be pounding at your door for a job, (2) searches that fail to include an overall assessment of the department in question are likely to miss the mark, (3) chairs must have demonstrated not only clinical expertise but also business savvy, (4) the best candidate is not necessarily someone who is already a department chair, (5) when it comes to chair searches, it's a buyer's market, and (6) the search process is inextricably linked to the success of the search. Key to the process of conducting an academic chair search are the judicious formation of the search committee; committee members' willingness to do their homework, attend all committee meeting, and keep the committee's activities confidential; crafting, not revising, the current job description for the open chair position; interviewing viable candidates rather than all candidates and adhering to a coordinated interviewing process; and evaluating internal and external candidates according to the same parameters. PMID- 12584090 TI - A computerized faculty time-management system in an academic family medicine department. AB - The authors describe the development, implementation, and evaluation of a computerized faculty time-management system (FTMS) in the Department of Family Medicine at the University of North Carolina-Chapel Hill. The FTMS is presented as an integrated set of computerized spreadsheets used annually to allocate faculty time across all mission activities of the department. It was first implemented in 1996 and has been continuously developed since then. An iterative approach has been used to gain consensus among faculty about time resources needed for various tasks of all missions of the department. These time-resource assumptions are used in the computerized system. Faculty time is allocated annually by the department vice chair in negotiation with individual faculty, making sure that the activities planned do not exceed the work time each faculty member has available for the year. During this process, faculty preferences are balanced against department aggregate needs to meet mission commitments and obligations. The authors describe how the computerized FTMS is used for faculty time management and career development, department planning, budget planning, clinical scheduling, and mission cost accounting. They also describe barriers and potential abuses and the challenge of building an organizational culture willing to discuss faculty time openly and committed to developing a system perceived as fair and accurate. The spreadsheet file is available free from the authors for use in other departments. PMID- 12584091 TI - Faculty development for educational leadership and scholarship. AB - The Medical Education Scholars Program (MESP) at the University of Michigan Medical School is designed to develop leaders in medical education. The program's goals are to enable faculty to provide curriculum direction, improved teaching, educational research, and development, and institutional leadership at all levels of medical education. This one-year program uses a variety of educational methods and provides a broad curriculum in educational theory, assessment and evaluation, research design and methods, teaching-skills development, and educational leadership. Faculty are admitted on a competitive basis and one half-day per week of release time is funded as part of the program. Salient outcomes of the program (promotions, educational research and development, curriculum leadership, and educational scholarship) were measured in a pre- and post-program design in which each scholar acted as his or her own control. There were major increases in promotions and educational awards, new educational responsibilities, and new educational programs. A particularly important outcome was the emergence of educational scholarship in the professional portfolios of the program scholars in the form of peer-reviewed presentations and publications and educational grant funding. A cost-outcome analysis indicates that these multi-year outcomes were obtained from a one-time investment of approximately $21,000 per graduating scholar. This evaluation indicates that intensive faculty development programs can have measurable impacts on the careers of the participants and the institutional environment. PMID- 12584092 TI - A teaching scholars program to develop leaders in medical education. AB - The authors describe a year-long faculty development initiative to develop leaders in medical education. The Teaching Scholars Program for Educators in the Health Sciences at McGill University enables faculty to improve their educational knowledge and skills while maintaining their clinical, teaching, and research responsibilities. The program, tailored to the participant's individual needs, consists of five main components: two university courses; independent study; participation in faculty-wide faculty development workshops and medical education rounds; a monthly seminar; and attendance at a national or international conference or course. Since its inception in 1997, 22 faculty members have completed the program; four are currently participating in it. This report discusses the experience of 15 scholars who completed the program by September 2000. Evaluations indicate that the scholars achieved most of their stated objectives. The university courses provided a foundation in educational principles and methodology; independent study allowed them to work on educational projects relevant to their disciplines; and the monthly seminars, faculty development workshops, and outside courses offered opportunities for skill acquisition and reflection. Participants also appreciated the opportunity to meet others interested in medical education and to become aware of available educational resources. A year after completing the program, many had joined new educational committees, taken on new leadership roles in medical education, and developed new courses for students and residents. Some of their projects had been presented at national meetings and were being prepared for publication. Two scholars had pursued further study at a master's level. Despite the ongoing challenge of protecting time for educational pursuits, this program has been beneficial in helping to develop educational leaders in the Faculty of Medicine. PMID- 12584093 TI - Bringing faculty development to community-based preceptors. AB - Community-based education of health profession students has increased dramatically, yet providing faculty development to a large, dispersed, and diverse population of community-based faculty is challenging. The authors describe lessons learned from 1997 to 2000 in developing, using, and disseminating a collection of preceptor development materials designed to be relevant to community-based faculty and easy to use. These activities were carried out by the Preceptor Development Program, which was developed by the Mountain Area Health Education Center of Asheville, North Carolina, which works with over 500 community preceptors of health profession students and medical residents. The program includes materials on nine core faculty development topics in a variety of formats: seminars, monographs, Web modules, and one-page summary "thumbnails." Faculty developers can download these free, customizable materials from the Web and are encouraged to adapt them for their own use. PMID- 12584094 TI - The impact of a program for systematically recognizing and rewarding academic performance. AB - PURPOSE: To describe an academic performance incentive system (APIS) and faculty perception of it; explore the impacts of incentive level, faculty rank, clinical practice volume, and administrative responsibility on academic productivity; and describe the APIS's use in maintaining congruence between department mission and activities. METHOD: A list of teaching, research, and academic service activities was developed, which full-time faculty (n = 33) used to report activities. Clinical faculty members received incentive income based on credits earned. APIS initially distributed 1% of practice plan receipts (subsequently increased to 3% and then 5%). Productivity was measured by differences in APIS points achieved. Satisfaction of all faculty participants was measured by survey. RESULTS: Faculty members (n = 20) who participated throughout averaged 22 credits per month (nine to 42 credits), and quarterly incentive bonuses ranged from 145 US dollars to 6,128 US dollars. Average credits earned per month were 24 for the 1% incentive, 23 for the 3% incentive, and 20 for the 5% incentive. Faculty members with administrative responsibilities were as productive academically as were their non administrative counterparts. Senior faculty members were as productive as junior faculty. Faculty members who were more productive clinically were more productive academically. Seventy percent of respondents reported they were either very satisfied or somewhat satisfied with the APIS. Seventy-eight percent felt that the APIS accurately reflected their academic productivity. Most respondents (81%) felt that the amount of money allocated to the incentive system was appropriate (15% felt it should be increased and one respondent recommended reduction). CONCLUSIONS: The APIS system has been well accepted by faculty and allows for data-driven discussion of the department's mission and activities. PMID- 12584095 TI - PhD faculty in clinical departments of U.S. medical schools, 1981-1999: their widening presence and roles in research. AB - PURPOSE: To analyze the growth, research activities, and academic status of PhD faculty in U. S. medical school clinical departments between 1981 and 1999. METHOD: Full-time U.S. medical school faculty who had PhDs and appointments in clinical departments in 1981 and 1999 and junior PhD faculty who became assistant professors between 1981 and 1990 were selected from the Faculty Roster System of the Association of American Medical Colleges. Their research activities and academic statuses were compared with those of MD or MD/PhD faculty in clinical departments or PhD faculty in basic science departments in the same years. RESULTS: The number of PhD faculty in clinical departments now exceeds the number of PhD faculty in basic science departments. PhD faculty in clinical departments come from diverse research backgrounds, contribute substantially to the research intensity of their institutions, and are more likely than their counterparts in basic science departments to become involved in research involving human participants or human tissues. PhD faculty in clinical departments are less likely than their counterparts in basic science departments, but are as likely as physicians in clinical departments, to be rewarded with academic promotion. They are less likely than their physician colleagues to be promoted in research intensive departments such as departments of medicine and at top 20 research intensive schools. CONCLUSIONS: The burgeoning career opportunities for PhD faculty in clinical departments should be reflected in the course work, mentorship, and potential thesis topics of PhD training programs. In lieu of tenure, research-intensive medical schools should develop alternative career tracks providing somewhat greater job stability for these faculty. PMID- 12584096 TI - Bridging postdoctoral training and a faculty position: initial outcomes of the Burroughs Wellcome Fund Career Awards in the Biomedical Siences. AB - PURPOSE: To describe the experiences of recipients of the Burroughs Wellcome Fund's Career Awards in the Biomedical Sciences Program (CABS) and highlight selected outcomes of the award's first set of graduates. METHOD: Since 1999, all active CABS recipients have been surveyed with the goal of assessing the program's implementation and the career progress of award recipients. Data were collected on multiple variables that are indicative of establishing an independent research career (e.g., time spent on research, external grant support, and scholarly publications), along with the views of individuals about the program and how the award had affected their careers. RESULTS: As evidenced by current employment and research funding, 77% of 1995-1999 grantees had been successful in securing tenure-track faculty positions in research-intensive institutions. Of those with faculty appointments, 78% had built research programs that attracted external support. In addition, 95% credited the award with facilitating their searches for faculty jobs, 70% believed that it had shortened the time required to obtain such positions, and 82% cited the award as enhancing their ability to attract research funds. They also perceived the award as allowing them to pursue research ideas that might have been considered as risky or premature by more traditional research sponsors. CONCLUSION: The CABS program has helped participants to launch careers as active and independent investigators. Grantees also credit it with allowing them to pursue research ideas that might otherwise have been considered impractical to support. PMID- 12584097 TI - Learning differences, medical students, and the law. AB - To update her article in the June 1999 issue of Academic Medicine, the author addresses the impact of the U.S. Supreme Court's decision in the 2001 Bartlett case on medical students with learning differences. In Bartlett v. the New York State Board of Bar Examiners, the Court ruled that Bartlett was substantially limited in the major life activity of working because of the board's failure to accommodate her reading impairment. The author postulates that the Supreme Court decision in the Bartlett case offers hope to medical students applying for accommodation on medical licensing examinations. If such accommodations are not forthcoming, she suggests that medical schools might ask the question, "Are board examinations a valid measurement of the preparation for the job of a physician?" PMID- 12584098 TI - Learning disabilities, professionalism, and the practice of medical education. AB - The authors use the interface of learning disabilities (LDs) and medical education to explore several issues relevant to medical professionalism and the training of future physicians. First, they examine arguments given by Little (in the preceding article) that a successful suit for accommodations on a state bar exam is generalizable to LDs and medical education, and suggest ways in which this may not be true. They then explore two frameworks for understanding medical education: (1) as a process of academic achievement linked to degree attainment, and (2) as a process of professional acculturation linked to competencies. Within this dichotomy, they then explore (1) the legitimacy accorded to different types of accommodations, (2) differing meanings of a "level playing field," and (3) the legal standard of "otherwise able." They also examine the use of intermediaries (e.g., a reader) as a "leveling" strategy and how, in clinical settings, this might violate core standards of autonomous decision making. The authors investigate the nature of "technical standards" in training across medicine and nursing and find a number of differences, particularly in the intent and levels of detail of standards. Across these two domains, they observe a status hierarchy, medical hubris, and the emergence of a "right to fail" as one travels down that hierarchy. The authors also examine medicine as an undifferentiated degree and consider arguments that medical school course requirements should be unbundled. They close by insisting that medical schools have a social responsibility to shift their pedagogic gaze from identifying handicaps in individuals to understanding how the education of physicians can become, quite literally, "handicapping." PMID- 12584100 TI - Getting acquainted with the medicines [excerpt]. PMID- 12584102 TI - A comparison of critical thinking in groups of third-year medical students in text, video, and virtual PBL case modalities. AB - PURPOSE: To determine whether critical thinking in problem-based learning (PBL) group discourse differed according to case modality. METHOD: The study was conducted in 2000 in the Department of Pediatrics at the University of Colorado School of Medicine in Denver. Third-year medical students on their pediatrics clerkship were divided into three groups: face-to-face with a text case, face-to face with a digital video case, and virtual with a digital video case. Twenty four groups were divided among the three case modalities. Using an existing coding scheme, each distinct codable unit of discourse was placed into one of 35 indicators reflective of five critical-thinking stages. For each group's discourse, a critical-thinking ratio was calculated for each of the stages. The Kruskal-Wallis test was used to compare the critical-thinking ratios for each stage across the three modalities. Residual conversation to indicate processes occurring within the group was also coded. RESULTS: A content analysis of the transcripts of 13 of the 24 group discussions occurred. The virtual groups had the highest critical-thinking ratio. Except for the problem-identification stage, the video groups had higher ratios that the text groups did. CONCLUSIONS: This exploratory study examined how the mode of case presentation affected the critical thinking of groups using the PBL format. Students who learned in a virtual modality with a digital video case engaged in more critical thinking. The data suggest that the video enhanced critical thinking in both face-to-face and virtual PBL groups. PMID- 12584103 TI - The roles and experiences of nonaffiliated and non-scientist members of institutional review boards. AB - PURPOSE: Little is known about the roles and experiences of non-scientist and nonaffiliated institutional review board (IRB) members (also known as lay members), and what contributions they are making to IRBs. This study investigated the experiences of IRB lay members from leading academic medical institutions in the United States and presents their recommendations for future education and training. METHOD: In 2000, the authors randomly selected and contacted 20 IRBs. From the 11 IRBs that agreed to participate, a total of 32 lay members participated in telephone interviews in which they were asked open-ended questions about the types of initial and ongoing education they had received, their interactions with scientific members, their contributions, the problems they experienced on the IRB, and recommendations for future education and training. RESULTS: Participants believed their role was to represent the community of human subjects, and 94% reported that their main contribution was simplifying the consent forms. Although 94% of participants had positive experiences working with scientist IRB members, 88% occasionally had been intimidated and felt disrespected by them. Forty-seven percent of participants identified lack of education and training as a problem, and 78% wanted more intensive education and training for future non-scientist/nonaffiliated members. CONCLUSION: IRB reform should include better training for non-scientist and nonaffiliated members so that they can take on more active roles. In addition, measures are needed to strengthen the relationships between scientist and non scientist and nonaffiliated members. PMID- 12584104 TI - Objectivity in objective structured clinical examinations: checklists are no substitute for examiner commitment. AB - PURPOSE: This study explored factors that contribute to objectivity in objective structured clinical examinations (OSCEs). The authors quantified the effect of examiners on interrater reliability and separated this effect from that of station construction, determined the effect of objectification on station reliability and validity, and explored examiner factors that may contribute to interrater reliability. METHOD: Data came from examiners' mark sheets from four annual OSCEs (1997-2000). The OSCEs were conducted identically and simultaneously at three sites, within the University of Otago medical school in New Zealand, with two examiners at each station. The contribution to interrater correlations of station construction and mark sheet compared with examiners' contribution was partitioned out using a random-effects analysis of variance. For one OSCE, a multiple linear regression was used to determine the independent contributions to interrater reliability of the number of checklist items per mark sheet, examiner experience, and examiner involvement in station construction. RESULTS: Station construction and mark sheets contributed 10.1% and examiners contributed 89.9% to the variation in interrater reliability. Following multivariate analysis, the number of items per mark sheet was negatively associated, and examiner involvement in station construction was positively associated, with interrater reliability. Examiner experience in examining or in clinical medicine was not associated with interrater reliability. There was a negative, but nonsignificant, correlation between number of items per mark sheet and that station's correlation with the aggregate OSCE mark. CONCLUSIONS: The contribution of objective mark sheets to objectivity is relatively minor compared with examiners' contribution. Increasing the number of checklist items per mark sheet decreased both reliability and validity. Achieving objectivity requires diligent examiners who are involved in the whole assessment. PMID- 12584105 TI - Housestaff-faculty ratios in ambulatory clinics and patient care and education. AB - This time-motion study examined relationships between housestaff-faculty ratios in a primary care clinic and measures of patient care and education. PMID- 12584106 TI - Protecting subjects, preserving trust, promoting progress I: policy and guidelines for the oversight of individual financial interests in human subjects research. AB - In December 2001, the AAMC Task Force on Financial Conflicts of Interest in Clinical Research released this report, the first of two (both published in this issue of Academic Medicine). This report focuses on gaps in existing federal financial disclosure regulations of individual conflicts of interests, finding that additional scrutiny is recommended in two areas: human subjects research and privately sponsored research. The task force suggests that when potential conflicts exist, a conflicts of interest committee should apply a rebuttable presumption against engaging in human subjects research. The task force recommends that the circumstances giving rise to the presumption against the proposed activity be balanced against compelling circumstances in favor of the conduct of the research. The AAMC task force delineates core principles to guide institutional policy development. First, an institution should regard all significant financial interests in human subjects research as requiring close scrutiny. Second, in the event of compelling circumstances, an individual holding a significant financial interest may be permitted to conduct the research. Whether circumstances are deemed compelling will depend in each case upon the nature of the science, the nature of the interest, how closely the interest is related to the research, and the degree to which the interest may be affected by the research. Four other core principles for development of institutional policies are identified in the report, pertaining to reporting, monitoring, management of conflicts, and accountability. PMID- 12584107 TI - Protecting subjects, preserving trust, promoting progress II: principles and recommendations for oversight of an institution's financial interests in human subjects research. AB - The AAMC Task Force on Financial Conflicts of Interest in Clinical Research issued this report, the second of two, in October 2002. (The first report is also published in this issue of Academic Medicine.) This report offers a unique perspective on the new phenomenon of "institutional" conflicts of interest. The task force acknowledges the diverse obligations of academic institutions that conduct research and also invest in-and accept the philanthropy of-commercial research sponsors. The task force emphasizes the importance of disclosing institutional financial interests as an integral part of the research process, critical to allaying public concerns, and to strengthening the trust relationship between research subjects, the public and the scientific community. The task force found that the safety and welfare of research subjects and the objectivity of the research could be-or could appear to be-compromised whenever an institution holds a significant financial interest that may be affected by the outcome of the research. Thus, the task force recommends separating the functional and administrative responsibilities related to human subjects research from those related to investment managing and technology licensing, and encourages the establishment of institutional conflicts-of-interest committees. As in the first report, the task force recommends that institutions should develop policies establishing a rebuttable presumption against the conduct of research at or under the auspices of an institution where potential conflicts in human subjects research are identified. This presumption against engaging in the research is to be balanced against compelling circumstances in favor of the conduct of the proposed research activity. PMID- 12584108 TI - Neuromedin U acts in the central nervous system to inhibit gastric acid secretion via CRH system. AB - Neuromedin U (NMU) is a hypothalamic peptide involved in energy homeostasis and stress responses. NMU, when administered intracerebroventricularly, decreases food intake and body weight while increasing body temperature and heat production. In addition, NMU, acting via the corticotropin-releasing hormone (CRH) system, induces gross locomotor activity and stress responses. We studied the effect of intracerebroventricularly administered NMU (0.5-4 nmol) in the regulation of gastric functions in conscious rats. Intracerebroventricular administration of NMU significantly decreased gastric acid output to 30-60% and gastric emptying to 35-70% in a dose-dependent manner. Vagotomy did not abolish the inhibitory effect of NMU on pentagastrin-induced gastric acid secretion. Pretreatment with indomethacin (10 mg/kg), an inhibitor of prostaglandin synthesis, also did not affect NMU-induced acid inhibition. Pretreatment with anti-CRH IgG (1 microg/rat), however, completely blocked NMU-induced acid inhibition (P < 0.01). Administration of yohimbine (4 mg/kg), an alpha(2) adrenergic receptor antagonist, also abolished NMU-induced acid inhibition (P < 0.01). These findings suggest that NMU is critical in the central regulation of gastric acid secretion via CRH. PMID- 12584109 TI - The effect of gastric electrical stimulation on canine gastric slow waves. AB - This study determined the most efficient parameters of low-frequency/long-pulse gastric electrical stimulation (GES) required to entrain gastric slow waves and also evaluated the effect of entrainment and high-frequency, short-pulse GES on gastric electrical activity (GEA). Nine dogs were fitted with stimulation wires along the greater curvature. Entrainment was observed in six or seven animals, with long-pulse GES at six cycles per minute (cpm), at various combinations of current and pulse width and was directly related to the energy delivered. Entrainment was observed in four to seven animals, with GES at 12 cpm, and the maximal driven frequency was 6 cpm. Entrainment did not significantly increase the dominant power of GEA. High-frequency, short-pulse GES, using pulse trains of 14 Hz, 5 mA, and 330 micros, with 0.1 s on and 5 s off, and pulse trains of 40 Hz, 10 mA, and 330 micros, with 2 s on 3 s off, did not affect variables of GEA. We conclude that acute low-frequency GES but not high-frequency, short-pulse GES can entrain slow waves; the power of slow waves is not affected by either type of stimulation. PMID- 12584110 TI - Melatonin-induced calcium signaling in clusters of human and rat duodenal enterocytes. AB - The amount of melatonin present in enterochromaffin cells in the alimentary tract is much higher than that in the central nervous system, and melatonin acting at MT(2) receptors mediates neural stimulation of mucosal HCO(3)(-) secretion in duodenum in vivo. We have examined effects of melatonin and receptor ligands on intracellular free calcium concentration ([Ca(2+)](i)) signaling in human and rat duodenal enterocytes. Clusters of interconnecting enterocytes (10-50 cells) were isolated by mild digestion (collagenase/dispase) of human duodenal biopsies or rat duodenal mucosa loaded with fura-2 AM and attached to the bottom of a temperature-controlled perfusion chamber. Clusters provided viable preparations and respond to stimuli as a syncytium. Melatonin and melatonin receptor agonists 2-iodo-N-butanoyl-5-methoxytryptamine and 2-iodomelatonin (1.0-100 nM) increased enterocyte [Ca(2+)](i), EC(50) of melatonin being 17.0 +/- 2.6 nM. The MT(2) receptor antagonists luzindole and N-pentanoyl-2-benzyltryptamine abolished the [Ca(2+)](i) responses. The muscarinic antagonist atropine (1.0 microM) was without effect on basal [Ca(2+)](i) and did not affect the response to melatonin. In the main type of response, [Ca(2+)](i) spiked rapidly and returned to basal values within 4-6 min. In another type, the initial rise in [Ca(2+)](i) was followed by rhythmic oscillations of high amplitude. Melatonin-induced enterocyte [Ca(2+)](i) signaling as well as mucosal cell-to-cell communication may be involved in stimulation of duodenal mucosal HCO(3)(-) secretion. PMID- 12584111 TI - Embolization by sinusoidal lining cells obstructs the microcirculation in rat sinusoidal obstruction syndrome. AB - Mechanisms leading to the obstruction of the microcirculation in sinusoidal obstruction syndrome (SOS) have been unclear. Because this occurs at the onset of disease, this is a potential key target for therapeutic intervention. Rats were treated with monocrotaline with or without continuous intraportal infusion of glutathione and were studied at 0.5, 1, 2, 4, 6, and 10 days after monocrotaline treatment with the use of in vivo microscopy and transmission electron microscopy. Sinusoidal perfusion decreased from days 1 through 10 with a nadir on day 4. At 12 h, numerous swollen sinusoidal endothelial cells (SECs) were observed. Subsequently, red blood cells penetrated into the space of Disse through gaps between and through swollen SEC and dissected the sinusoidal lining away from the parenchymal cells. Sinusoidal blood flow was obstructed by an embolism of aggregates of sinusoidal lining cells, red blood cells, and adherent monocytes. All changes were prevented by glutathione infusion, notably the initial swelling of SEC. SOS is initiated by changes in SEC. Microcirculatory obstruction is due to dissection of the sinusoidal lining, followed by embolization of the sinusoid by sinusoidal lining cells, compounded by aggregates of monocytes adherent in the sinusoids. Glutathione prevents SOS by preserving an intact sinusoidal barrier. PMID- 12584112 TI - S-nitrosothiols inhibit uterine smooth muscle cell proliferation independent of metabolism to NO and cGMP formation. AB - S-nitrosothiols (RSNOs) are important mediators of nitric oxide (NO) biology. The two mechanisms that appear to dominate in their biological effects are metabolism leading to the formation of NO and S-nitrosation of protein thiols. In this study we demonstrate that RSNOs inhibit uterine smooth muscle cell proliferation independent of NO. The antiproliferative effects of NO on vascular smooth muscle are well defined, with the classic NO-dependent production of cGMP being demonstrated as the active pathway. However, less is known on the role of NO in mediating uterine smooth muscle cell function, a process that is important during menstruation and pregnancy. The RSNOs S-nitrosoglutathione and S-nitroso-N-acetyl pencillamine inhibited growth factor-dependent proliferation of human and rat uterine smooth muscle cells (ELT-3). Interestingly, these cells reduced RSNOs to generate NO. However, use of NO donors and other activators of the cGMP pathway failed to inhibit proliferation. These findings demonstrate the tissue-specific nature of responses to NO and demonstrate the presence of a RSNO-dependent but NO independent pathway of inhibiting DNA synthesis in uterine smooth muscle cells. PMID- 12584113 TI - Neurotensin stimulates IL-8 expression in human colonic epithelial cells through Rho GTPase-mediated NF-kappa B pathways. AB - Neurotensin (NT), a neuropeptide highly expressed in the gastrointestinal tract, participates in the pathophysiology of intestinal inflammation. We recently showed that NT stimulates interleukin-8 (IL-8) expression in NCM460 nontransformed human colonic epithelial cells via both mitogen-activating protein kinase (MAPK)- and NF-kappaB-dependent pathways. However, the molecular mechanism by which NT induces expression of proinflammatory cytokines such as IL-8 has not been investigated. In this study we show that inhibition of endogenous Rho family proteins (RhoA, Rac1, and Cdc42) by their respective dominant negative mutants inhibits NT-induced IL-8 protein production and promoter activity. Western blot experiments demonstrated that NT strongly activated RhoA, Rac1, and Cdc42. Overexpression of the dominant negative mutants of RhoA, Rac1, and Cdc42 significantly inhibited NT-induced NF-kappaB-dependent reporter gene expression and NF-kappaB DNA binding activity. NT also stimulated p38 MAPK phosphorylation, and overexpression of dominant negative mutants of RhoA, Rac1, and Cdc42 did not significantly alter p38 and ERK1/2 phosphorylation in response to NT. Together, our findings indicate that NT-stimulated IL-8 expression is mediated via a Rho dependent NF-kappaB-mediated pathway. PMID- 12584114 TI - Autoregulation of glucocorticoid receptor by cortisol in rainbow trout hepatocytes. AB - We used primary cultures of trout hepatocytes and a physiological dose of cortisol (100 ng/ml), mimicking stressed levels in salmonid fish, to address the impact of glucocorticoid stimulation on glucocorticoid receptor (GR) mRNA abundance and protein content. Cortisol significantly elevated GR mRNA content over a 24-h period; this increase was abolished by actinomycin D, suggesting transcriptional control of GR. However, cortisol significantly decreased GR protein content, leading us to hypothesize that lower GR protein content may be regulating GR mRNA abundance. Indeed, treatment of hepatocytes with MG-132, a proteasomal inhibitor shown to prevent GR degradation by cortisol, abolished cortisol-mediated GR mRNA upregulation. Also, geldanamycin, a heat shock protein 90-specific inhibitor, abolished the GR mRNA increase evident with cortisol but did not modify cortisol-induced increases in abundance of mRNA for phosphoenolpyruvate carboxykinase, a glucocorticoid-responsive gene, or hepatocyte glucose release. Together, our results suggest a negative feedback loop for GR gene regulation by cortisol in trout hepatocytes. The autoregulation of GR may be a crucial step in the physiological stress response process, especially in modulating energy-dependent processes that are glucocorticoid dependent, including gluconeogenesis. PMID- 12584115 TI - Two-photon molecular excitation imaging of Ca2+ transients in Langendorff perfused mouse hearts. AB - The ability to image calcium signals at subcellular levels within the intact depolarizing heart could provide valuable information toward a more integrated understanding of cardiac function. Accordingly, a system combining two-photon excitation with laser-scanning microscopy was developed to monitor electrically evoked [Ca(2+)](i) transients in individual cardiomyocytes within noncontracting Langendorff-perfused mouse hearts. [Ca(2+)](i) transients were recorded at depths or =1 mM, increasing linearly with substrate concentration. CYP2E1 was the primary enzyme involved in the reductive reaction, as univocally indicated by all the different approaches. The reductive pathway seems to be scarcely relevant in the human liver, since it is active only at high substrate concentrations, and in strictly anaerobic conditions. The role of human CYP2E1 in CHCl(3) metabolism at low levels, typical of actual human exposure, provides insight into the molecular basis for eventual difference in susceptibility to chloroform-induced effects due to either genetic, pathophysiological, or environmental factors. PMID- 12584153 TI - Oxidative metabolism of amprenavir in the human liver. Effect of the CYP3A maturation. AB - Amprenavir is a human immunodeficiency virus-1 (HIV-1) protease inhibitor intended to be used to treat HIV-infected children. Although a pediatric dosage is proposed by the manufacturer, no data are currently available on the pharmacokinetics of amprenavir in neonates and infants. Amprenavir being primarily eliminated after oxidative biotransformation, we explored its in vitro metabolism by cytochrome P450 (P450)-dependent monooxygenases. In our conditions, five metabolites were formed in vitro and subsequently analyzed by liquid chromatography-mass spectrometry; P450-dependent oxidations occurred either on the tetrahydrofuran ring (M3 and M4), the aniline ring (M5), and the aliphatic chain (M2) or resulted from the N-dealkylation and loss of the tetrahydrofuran ring (M1). The two major metabolites, respectively M3 and M2 were formed by human liver microsomes with K(m) between 10 and 70 microM. CYP3A4 and to a lesser extent CYP3A5 were major contributors for the formation of M2, M3, and M5 metabolites, whereas CYP3A7 had no or little activity. This assumption was confirmed by inhibition with ketoconazole and ritonavir (two potent inhibitors of CYP3A) whereas sulfaphenazole (2C9 inhibitor) and quinidine (2D6 inhibitor) were inefficient. The metabolism of amprenavir was negligible in microsomes from either fetuses or neonates and steadily increased after the first weeks of life in relation with the maturation of CYP3A4/5. In conclusion, results demonstrated that the capacity of the human liver to oxidize amprenavir is low during the first weeks after birth and that dosage could be substantially reduced during the early neonatal period. PMID- 12584154 TI - Induction of cytochrome P450 enzymes in cultured precision-cut human liver slices. AB - Precision-cut human liver slices obtained from 11 donors were cultured for 72 h in a defined medium (serum free Williams' medium E) supplemented with 0.1 microM insulin and 0.1 microM dexamethasone (DEX). Liver slices were treated with 50 microM concentrations of beta -naphthoflavone (BNF), lansoprazole, rifampicin (RIF), DEX and methylclofenapate and 500 microM sodium phenobarbital (NaPB). The relative apoprotein levels of 12 cytochrome P450 (P450) enzymes were determined in liver slice microsomes using a panel of antipeptide antibodies. Treatment with BNF significantly induced mean levels of CYP1A2 apoprotein to 160% of levels in 72-h control (no test compound) human liver slice microsomes. NaPB significantly induced levels of CYP3A4 apoprotein to 255% of control and RIF significantly induced levels of CYP2C19 and CYP3A4 apoproteins to 265 and 330% of control, respectively. In addition, treatment with RIF increased levels of CYP2A6 apoprotein to 205% of control, and treatment with both NaPB and RIF increased levels of CYP2B6 apoprotein to 370 and 615% of control, respectively. However, these increases were not statistically significant, owing to a variable response between liver slice preparations from different subjects, this being apparent for all inducible P450s. In contrast, none of the compounds examined significantly increased levels of CYP2C8, CYP2C9, CYP2D6, CYP2E1, and CYP4A11 apoproteins. Levels of CYP1A1 apoprotein were not detected in any liver slice sample, either before or after treatment with the model inducers. Overall, these results demonstrate the utility of cultured human liver slices for assessing the effects of chemicals on P450 enzymes. PMID- 12584155 TI - Apparent mechanism-based inhibition of human CYP2D6 in vitro by paroxetine: comparison with fluoxetine and quinidine. AB - Paroxetine, a selective serotonin reuptake inhibitor, is a potent inhibitor of cytochrome P450 2D6 (CYP2D6) activity, but the mechanism of inhibition is not established. To determine whether preincubation affects the inhibition of human liver microsomal dextromethorphan demethylation activity by paroxetine, we used a two-step incubation scheme in which all of the enzyme assay components, minus substrate, are preincubated with paroxetine. The kinetic parameters of inhibition were also estimated by varying the time of preincubation as well as the concentration of inhibitor. From these data, a Kitz-Wilson plot was constructed, allowing the estimation of both an apparent inactivator concentration required for half-maximal inactivation (K(I)) and the maximal rate constant of inactivation (k(INACT)) value for this interaction. Preincubation of paroxetine with human liver microsomes caused an approximately 8-fold reduction in the IC(50) value (0.34 versus 2.54 microM). Time-dependent inhibition was demonstrated with an apparent K(I) of 4.85 microM and an apparent k(INACT) value of 0.17 min(-1). Spectral scanning of CYP2D6 with paroxetine yielded an increase in absorbance at 456 nm suggesting paroxetine inactivation of CYP2D6 via the formation of a metabolite intermediate complex. This pattern is consistent with the metabolism of the methylenedioxy substituent in paroxetine; such substituents may produce mechanism-based inactivation of cytochrome P450 enzymes. In contrast, quinidine and fluoxetine, both of which are inhibitors of CYP2D6 activity, did not exhibit a preincubation-dependent increase in inhibitory potency. These data are consistent with mechanism-based inhibition of CYP2D6 by paroxetine but not by quinidine or fluoxetine. PMID- 12584156 TI - Absorption, distribution, metabolism, and excretion of ximelagatran, an oral direct thrombin inhibitor, in rats, dogs, and humans. AB - The absorption, metabolism, and excretion of the oral direct thrombin inhibitor, ximelagatran, and its active form, melagatran, were separately investigated in rats, dogs, and healthy male human subjects after administration of oral and intravenous (i.v.) single doses. Ximelagatran was rapidly absorbed and metabolized following oral administration, with melagatran as the predominant compound in plasma. Two intermediates (ethyl-melagatran and OH-melagatran) that were subsequently metabolized to melagatran were also identified in plasma and were rapidly eliminated. Melagatran given i.v. had relatively low plasma clearance, small volume of distribution, and short elimination half-life. The oral absorption of melagatran was low and highly variable. It was primarily renally cleared, and the renal clearance agreed well with the glomerular filtration rate. Ximelagatran was extensively metabolized, and only trace amounts were renally excreted. Melagatran was the major compound in urine and feces after administration of ximelagatran. Appreciable quantities of ethyl-melagatran were also recovered in rat, dog, and human feces after oral administration, suggesting reduction of the hydroxyamidine group of ximelagatran in the gastrointestinal tract, as demonstrated when ximelagatran was incubated with feces homogenate. Polar metabolites in urine and feces (all species) accounted for a relatively small fraction of the dose. The bioavailability of melagatran following oral administration of ximelagatran was 5 to 10% in rats, 10 to 50% in dogs, and about 20% in humans, with low between-subject variation. The fraction of ximelagatran absorbed was at least 40 to 70% in all species. First-pass metabolism of ximelagatran with subsequent biliary excretion of the formed metabolites account for the lower bioavailability of melagatran. PMID- 12584157 TI - Identification of trichloroethylene and its metabolites in human seminal fluid of workers exposed to trichloroethylene. AB - We have investigated the potential of the male reproductive tract to accumulate trichloroethylene (TCE) and its metabolites, including chloral, trichloroethanol (TCOH), trichloroacetic acid (TCA), and dichloroacetic acid (DCA). Human seminal fluid and urine samples from eight mechanics diagnosed with clinical infertility and exposed to TCE occupationally were analyzed. In in vivo experimental studies, TCE and its metabolites were determined in epididymis and testis of mice exposed to TCE (1000 ppm) by inhalation for 1 to 4 weeks. In other studies, incubations of monkey epididymal microsomes were performed in the presence of TCE and NADPH. Our results showed that seminal fluid from all eight subjects contained TCE, chloral, and TCOH. DCA was present in samples from two subjects, and only one contained TCA. TCA and/or TCOH were also identified in urine samples from only two subjects. TCE, chloral, and TCOH were detected in murine epididymis after inhalation exposure with TCE for 1 to 4 weeks. Levels of TCE and chloral were similar throughout the entire exposure period. TCOH levels were similar at 1 and 2 weeks but increased significantly after 4 weeks of TCE exposure. Chloral was identified in microsomal incubations with TCE in monkey epididymis. CYP2E1, a P450 that metabolizes TCE, was localized in human and monkey epididymal epithelium and testicular Leydig cells. These results indicated that TCE is metabolized in the reproductive tract of the mouse and monkey. Furthermore, TCE and its metabolites accumulated in seminal fluid, and suggested associations between production of TCE metabolites, reproductive toxicity, and impaired fertility. PMID- 12584158 TI - P-glycoprotein limits the brain penetration of nonsedating but not sedating H1 antagonists. AB - The present study evaluates the impact of P-glycoprotein (P-gp) on plasma-brain disposition and transepithelial transport of sedating versus nonsedating H1 antagonists using multidrug-resistant (mdr) gene 1a and 1b (mdr1a/b) knockout (KO) mice and human MDR1-transfected Madin-Darby canine kidney (MDCK) cells. Three nonsedating (cetirizine, loratadine, and desloratadine) and three sedating (diphenhydramine, hydroxyzine, and triprolidine) H1-antagonists were tested. Each compound was administered to KO and wild-type (WT) mice intravenously at 5 mg/kg. Plasma and brain drug concentrations were determined by liquid chromatography mass spectrometry analysis. Mean pharmacokinetic parameters (CL, V(ss), and t(1/2)) were obtained using WinNonlin. In addition, certirizine, desloratadine, diphenhydramine, and triprolidine (2 microM) were tested as substrates for MDR1 using MDR1-MDCK cells. The bidirectional apparent permeability was determined by measuring the amount of compound at the receiving side at 5 h. The brain-to plasma area under the curve (AUC) ratio was 4-, 2-, and >14-fold higher in KO compared with WT mice for cetirizine, loratadine, and desloratadine, respectively. In contrast, the brain-to-plasma AUC ratio between KO and WT was comparable for hydroxyzine, diphenhydramine, and triprolidine. Likewise, the efflux ratio between basolateral to apical and apical to basolateral was 4.6- and 6.6-fold higher in MDR1-MDCK than the parental MDCK for certirizine and desloratadine, respectively, whereas it was approximately 1 for diphenhydramine and triprolidine. Our results demonstrate that sedating H1-antagonists hydroxyzine, diphenhydramine, and triprolidine are not P-gp substrates. In contrast, nonsedating H1-antagonists cetirizine, loratadine, and desloratadine are P-gp substrates. Affinity for P-gp at BBB may explain the lack of central nervous system side effects of modern H1-antagonists. PMID- 12584159 TI - Effect of efflux inhibition on brain uptake of itraconazole in mice infected with Cryptococcus neoformans. AB - Itraconazole is a fungistatic agent that, although highly lipophilic, shows poor transport through the blood brain barrier that may be due to efflux proteins. The combined administration of an efflux inhibitor with itraconazole should increase cerebral itraconazole concentrations and therefore, improve the treatment of Cryptococcus neoformans meningitis with this antifungal agent. To test this hypothesis, we have studied the influence of murine cerebral infection with C. neoformans and the inhibition of efflux by intraperitoneal injection of a P glycoprotein inhibitor, GF120918 [N-(4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2 isoquinolinyl)-ethyl]-phenyl)9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamide], on the pharmacokinetics of itraconazole in plasma and brain after a single intraperitoneal itraconazole injection. We also investigated the influence of efflux inhibition on the efficacy of repeated doses of itraconazole in this murine model. The results showed that in healthy and infected mice pretreated or not with GF120918, plasma itraconazole values of area under the curve (AUC) were similar. In contrast, cerebral values of AUC were higher in infected mice compared with healthy mice. Moreover, the pretreatment of infected mice with GF120918 significantly increased cerebral itraconazole values of area under the curve and decreased weight loss in the treatment with itraconazole of a cerebral infection with C. neoformans. PMID- 12584160 TI - Tissue mRNA expression of the rat UDP-glucuronosyltransferase gene family. AB - UDP-Glucuronosyltransferases (UGTs) are phase II biotransformation enzymes that glucuronidate numerous endobiotic and xenobiotic substrates. Glucuronidation increases the water solubility of the substrate and facilitates renal and biliary excretion of the resulting glucuronide conjugate. UGTs have been divided into two gene families, UGT1 and UGT2. Tissue distribution of UGTs has not been thoroughly examined, and such data could provide insight into the importance of individual UGT isoforms in specific tissues and to the pharmacokinetics and target organ toxicity of UGT substrates. Therefore, the aim of this study was to determine mRNA levels of rat UGT1 and UGT2 family members in liver, kidney, lung, stomach, duodenum, jejunum, ileum, large intestine, cerebellum, and cerebral cortex, as well as nasal epithelium for UGT2A1. Tissue levels of UGT mRNA were detected using branched DNA signal amplification analysis. Three UGT isoforms, UGT1A1, UGT1A6, and UGT2B12, were detected in many tissues, whereas distribution of other UGT isoforms was more tissue-specific. For example, UGT2A1 was detected predominantly in nasal epithelium. Additionally, UGT1A5, UGT2B1, UGT2B2, UGT2B3, and UGT2B6 were detected primarily in liver. Furthermore, detection of UGT1A2, UGT1A3, UGT1A7, and UGT2B8 was somewhat specific to gastrointestinal (GI) tract. However, not all of these UGTs were detected in all portions of the GI tract. UGT1A8 was unique in that it was barely detectable in any of the tissues examined. In conclusion, some UGT isoforms were expressed in multiple tissues, whereas other UGT isoforms were predominantly expressed in a certain tissue such as nasal epithelium, liver, or GI tract. PMID- 12584161 TI - Metabolism and disposition of the antihypertensive agent moxonidine in humans. AB - The metabolism and pharmacokinetics of moxonidine, a potent central-acting antihypertensive agent, were studied in four healthy subjects after a single oral administration of approximately 1 mg (approximately 60 muCi) of [(14)C(3)]moxonidine. Moxonidine was rapidly absorbed, with peak plasma concentration achieved between 0.5 to 2 h postdose. The maximal plasma concentration and the area under the curve of unchanged moxonidine are lower than those determined for radioactivity, indicating presence of circulating metabolite(s). The total recovery of radiocarbon over 120 h ranged from 99.6 to 105.2%, with 92.3 to 103.3% of the radioactivity excreted in the urine and only 1.9 to 7.3% of the dose excreted in the feces. Thus, renal elimination represented the principal route of excretion of radioactivity. Metabolites of moxonidine were identified in urine and plasma samples by high performance liquid chromatography and liquid chromatography-tandem mass spectrometry. Oxidation of moxonidine on the methyl group or on the imidazoline ring resulted in the formation of hydroxymethyl moxonidine, hydroxy moxonidine, dihydroxy moxonidine, and dehydrogenated moxonidine. Metabolite profiling results indicated that parent moxonidine was the most abundant component in the urine. The dehydrogenated moxonidine was the major urinary metabolite as well as the major circulating metabolite. Moxonidine also underwent phase II metabolism, generating a cysteine conjugate. In summary, moxonidine is well absorbed after oral administration. The major clearance pathway for moxonidine in humans is via renal elimination. Furthermore, seven metabolites were identified with three metabolites unique to humans. PMID- 12584162 TI - The discovery of a new family of mammalian enzymes for repair of oxidatively damaged DNA, and its physiological implications. AB - Oxidatively damaged bases in the genome are likely to be responsible for mutations leading to sporadic carcinogenesis. Two structurally similar DNA glycosylases, NTH1 and OGG1, which are able to excise most of these damaged bases, were identified previously in mammalian cells. A distinct family, consisting of two human DNA glycosylases orthologous to enzymes in Escherichia coli, has recently been characterized; they have overlapping substrate ranges with NTH1 and OGG1. The presence of multiple enzymes with potential back-up functions underscores the importance of removing both endogenously and exogenously generated oxidatively damaged bases from the genome, and may explain why no cancer or other disease phenotype has so far been linked to the deficiency of a single DNA glycosylase. PMID- 12584163 TI - Mechanisms by which DNA tumor virus oncoproteins target the Rb family of pocket proteins. AB - Small DNA tumor viruses have evolved different mechanisms to abrogate the function of the retinoblastoma tumor suppressor (pRb). Studies of these viruses have been invaluable in uncovering the central role of the Rb family of pocket proteins in cell cycle control. While the molecular mechanisms by which the viral oncoproteins inactivate the Rb family are still being elucidated, it is clear that targeting of this family is required both for viral replication and for virus-induced transformation of mammalian cells. This review compares and contrasts the approaches DNA tumor viruses have evolved to antagonize Rb family members--ranging from relatively simple equilibrium dissociation of pRb from cellular pRb-binding factors to chaperone-mediated alterations in pocket protein stability and phosphorylation levels. The review will focus on the viral oncoproteins adenovirus E1A, human papillomavirus E7 and the large T antigens of several polyomaviruses. An understanding of these mechanisms may provide further insight into the regulation and functions of Rb family members as well as uncover new targets for the development of novel anti-viral agents, particularly against human papillomavirus, which is a significant cause of human cancer. PMID- 12584164 TI - Enhanced sensitivity of human oral tumours to the flavonol, morin, during cancer progression: involvement of the Akt and stress kinase pathways. AB - Various naturally occurring flavonoids have been found to be cancer-protective in chemically induced animal cancer models and synthetic flavonoid derivatives are being tested for potential chemotherapeutic usefulness in clinical trials. This report demonstrates that human oral squamous carcinoma cells (SCC) are significantly more sensitive to growth inhibition by the naturally occurring flavonoid, morin (3,5,7,2',4'-pentahydroxyflavone) than normal oral mucosa (NOMC) (SCC IC(50) = 115 microM; NOMC IC(50) = 173 micro M; P for difference = 0.009). Structure/function comparisons indicate that both the 2' and 4' hydroxyl groups in morin are required for its tumour selectivity. Morin causes growth arrest in G(2)/M, without inducing apoptosis, and this is associated with induction of GADD45 and phosphorylation and inactivation of the cell cycle kinase, cdc2. Morin also has pleiotropic effects on kinase signalling pathways, including inhibition of activation of protein kinase B by mitogens (but not extracellular-regulated kinases 1/2) and activation of the stress pathway kinases, Jun N-terminal kinase and p38 kinase. p38 kinase activation is functionally important since inhibition of its activation by the specific inhibitor SB202190 partially prevented cell cycle arrest by morin. However, analysis of dose-response relationships reveals that the enhanced tumour sensitivity to morin may be explained by the fact that activation of AKT is inhibited at lower concentrations of morin in carcinomas than normal oral mucosa, whereas Jun N-terminal kinase, p38 kinase and GADD45 are all induced in parallel with the same dose-response curves in carcinomas and normal oral mucosa. PMID- 12584165 TI - Minimal effects of dietary restriction on neuroendocrine carcinogenesis in Rb+/- mice. AB - The efficacy of dietary restriction in retarding tumor growth is well established in rodents. However, gene and cell lineage specificity of dietary restriction effects is far less defined. Mice with a single copy of the retinoblastoma susceptibility gene (Rb) develop a well-established syndrome of mouse neuroendocrine neoplasia associated with Rb deficiency. Thus, if DR represses tumor growth in this model, it should be unambiguously attributed to the Rb defect in neuroendocrine cell lineages. To address this possibility, Rb(+/-) mice were entered into a diet restriction study. Surprisingly, 40-50% reductions in dietary intake, relative to an ad libitum group, started on either postnatal day 28 or 42 had little to no effect on either the frequency or growth of pituitary tumors either during the latency period (postnatal day 224) or at the time of their natural death. Consistent with cross-section data, survival of 65 diet restricted Rb(+/-) mice was almost identical to that of 67 Rb(+/-) mice fed ad libitum (AL); median life span was 414 and 436 days for AL and DR groups, respectively. These findings indicate that diet restriction provides no significant benefit in delaying growth and progression of neuroendocrine tumors exhibiting loss of RB function. They also introduce the possibility that RB is required for the tumor-repressive effects of DR. PMID- 12584166 TI - All-trans retinoic acid (atRA) differentially induces apoptosis in matched primary and metastatic melanoma cells -- a speculation on damage effect of atRA via mitochondrial dysfunction and cell cycle redistribution. AB - All-trans retinoic acid (atRA) has been suggested to exert its cytotoxicity via apoptosis but the mechanisms behind the damage effects have not been fully understood. In this study, we investigated the cytotoxic effects of atRA in eleven primary and matched metastatic cutaneous melanoma cell lines. All the primary and metastatic melanoma cell lines examined expressed the retinoic acid receptors. The cultured melanoma cells treated with atRA showed dysfunction of mitochondria and altered cell cycle distribution, inhibited cell proliferation and apoptosis. The cytotoxic effects of atRA were dose- and time-dependent. The dysfunction of mitochondria and induction of apoptosis were more pronounced in the primary tumor cells than in the metastatic cell lines from the same patients. The data indicate that the cytotoxic effect of atRA was mediated through dysfunction of mitochondria, alterations in cell cycle and induction of apoptosis. Melanoma in early stage may have better response to atRA adjuvant therapy than the melanoma in late stage, suggesting the early utility of atRA in melanoma chemotherapy. PMID- 12584167 TI - Molecular progression of promoter methylation in intraductal papillary mucinous neoplasms (IPMN) of the pancreas. AB - To understand the role of gene promoter methylation in neoplastic evolution and progression, the methylation changes associated with 15 candidate tumor suppressor genes were studied throughout stages of tumor progression involving intraductal papillary mucinous neoplasms (IPMN) of the pancreas. Genomic DNA from 28 pancreatic IPMN tissue samples, categorized histologically as non-invasive intraductal IPMN (n = 3), IPMN with carcinoma in situ (n = 7), IPMN with microinvasion <1 mm (n = 4), and infiltrative IPMN with associated adenocarcinoma (n = 14), was modified by bisulfite treatment and analyzed with methylation specific PCR (MSP). Promoter methylation of at least one tumor suppressor gene was present in 26/28 (92%) of the IPMNs. The cell cycle control genes, p16 and p73, were methylated frequently (>50%) in both non-invasive and invasive tumors. APC methylation was discovered in <10% of the non-invasive IPMNs versus 45% of the IPMNs associated with infiltrative adenocarcinoma, P = 0.040. Mismatch repair genes, hMLH1 and MGMT, were frequently methylated in the invasive IPMNs compared with the non-invasive tumors (38 versus 10% and 45 versus 20%, respectively) as was E-cadherin (38 versus 10%), P = 0.11. Multiple gene methylation at greater than three loci was present in 55% of the invasive tumors compared with 20% of the non-invasive tumors, P = 0.075. Lymph node status did not predict multi-gene methylation among tumors associated with invasive cancer. Compared with non invasive IPMNs of the pancreas, IPMNs associated with adenocarcinoma demonstrate higher rates of aberrant tumor suppressor gene methylation. The sequential acquisition of hypermethylation at multiple gene promoter sites may explain tumor progression in IPMNs and other malignancies. Detection of methylation within selected genes may afford an accurate diagnostic molecular marker and predictor of neoplastic behavior. PMID- 12584168 TI - Thrombospondin 1--a regulator of adenoma growth and carcinoma progression in the APC(Min/+) mouse model. AB - Thrombospondin 1 (TSP-1) is a multifunctional extracellular matrix protein that is an endogenous regulator of tumor angiogenesis. The effects of TSP-1 on adenoma formation and development into cancerous lesions has been evaluated in the Min(/+) (multiple intestinal neoplasia) mouse model. These mice develop multiple adenomas in the small intestine due to a mutation in the homologous APC (adenomatous polyposis coli) gene. As in its human counterpart, these adenomas may progress to carcinomas. Intestines of APC(Min/+) mice were dissected and histologic evaluation of adenomas was then conducted. Significant increases in vascularization and proliferation were observed in adenomatous, as compared with normal, mucosa. TSP-1 immunostaining revealed significant decreases in the number and intensity of positive cells in adenomas, as compared with normal mucosa. TSP 1 scores were inversely correlated with vascularity and proliferation rate. Cross breeding of mice homozygous for a deletion of the TSP-1 gene (TSP-1(-/-)) with mice heterozygous for the APC gene mutation (APC(Min/+)), resulted in animals that showed a significant increase in adenoma number and diameter. Also, histopathological examination of these adenomas showed accelerated dysplasic changes, carcinoma in situ and early invasion, compared with their APC(Min/+) littermates. Moreover, a significant decrease of TUNEL-positive cells was observed in intestinal adenomas of TSP-1(-/-)/APC(Min/+) mice. This study reports the first in vivo impact of TSP-1 during early stages of tumor initiation and development in an intestinal carcinogenesis model and demonstrates that TSP-1 affects both angiogenesis and tumor cell apoptosis. PMID- 12584169 TI - 2-Methoxyestradiol interferes with NF kappa B transcriptional activity in primitive neuroectodermal brain tumors: implications for management. AB - Medulloblastoma (MB) is a primitive neuroectodermal tumor (PNET) of the central nervous system (CNS) and the most common malignant primary brain tumor in children. Currently, poor risk and recurrent MB patients are treated with cytotoxic chemotherapy alone or in combination with surgery and irradiation. In order to improve on therapeutic outcome and reduce toxicity of current treatment strategies, new and novel therapeutic agents are needed for MB patients. To that purpose, we have examined the effect of 2-methoxyestradiol (2-ME), an endogenous non-toxic estrogenic metabolite on the growth of three medulloblastoma cell lines (DAOY, D341 and D283); and two high-grade anaplastic astrocytoma/glioblastoma cell lines, U-87MG and T-98-G. We present evidence to show that 2-ME preferentially inhibits the growth of medulloblastoma cells significantly by blocking cell cycle progression predominantly in G(2)/M phase. 2-ME treatment results in phosphorylation of cdc25C without any significant alterations in the expression of cyclin B1 or p34cdc2. In addition, we observed a decrease in the levels of 14-3-3 proteins following treatment with 2-ME. Furthermore, 2-ME mediated growth inhibition is accompanied by induction of apoptosis as evidenced by morphological alterations and DNA fragmentation analysis. Of interest is the finding that 2-ME induced apoptosis is not mediated through alterations in the expression of p53 or Bax and that transcriptional activity of NF kappa B and DNA binding activity is reduced indicating that 2-ME disrupts the NF kappa B signaling pathway. These results suggest that 2-ME may prove to be a useful therapeutic agent in the treatment of PNET brain tumors such as medulloblastoma. In addition, as 2-ME inhibits growth predominantly through G(2)/M block, it may enhance the effectiveness of radiation therapy. PMID- 12584170 TI - Msh2 deficiency enhances somatic Apc and p53 mutations in Apc+/-Msh2-/- mice. AB - Inactivation of the adenomatous polyposis coli (Apc) gene by loss of the wild type Apc allele (LOH) is a prerequisite for the development of intestinal adenomas in Msh2 proficient Min (Apc+/-Msh2+/+) mice. In contrast, adenomas from Msh2 deficient Min (Apc+/-Msh2-/-) mice are not usually associated with LOH. Given the role of Msh2 in post-replicative DNA repair, this study investigated whether Msh2 deficiency enhances somatic Apc and p53 mutations in Apc+/-Msh2-/- mice. Somatic Apc mutations (5/sample) were observed in the non-neoplastic intestinal mucosa from Apc+/-Msh2-/- mice but not from Min mice, suggesting that Msh2 deficiency is associated with a hypermutable state in the intestinal mucosa from Apc+/-Msh2-/- mice. Adenomas from Apc+/-Msh2-/- mice had a 2-fold higher rate of somatic Apc mutations (10/adenoma) than the non-neoplastic intestinal mucosa (5/sample), and did not demonstrate LOH. Truncating Apc mutations were observed in 82% of the adenomas from Apc+/-Msh2-/- mice and were not observed at all in the non-neoplastic intestinal mucosa. In contrast, in Min mice, all adenomas demonstrated LOH, had significantly less numbers of somatic Apc mutations (1.8 mutations/adenoma) compared with the adenomas from Apc+/-Msh2-/- mice, and harbored no truncating Apc mutations. These observations suggest that somatic Apc mutations, and not LOH, is a likely mechanism by which the Apc gene is inactivated in the development of adenomas in Apc+/-Msh2-/- mice in contrast to Min mice. Adenomas from Apc+/-Msh2-/- mice, but not from Min mice, also harbored somatic p53 mutations (mutation frequency of 45.5%), reflecting hypermutability associated with Msh2 deficiency. The nature and frequency of somatic Apc and p53 mutations in Apc+/-Msh2-/- mice suggest that many genomic sites, in addition to genes containing simple repeated sequences, are at risk of somatic mutations associated with Msh2 deficiency. PMID- 12584171 TI - Response of human mammary epithelial cells to DNA damage induced by BPDE: involvement of novel regulatory pathways. AB - The responses of a line of normal human mammary epithelial cells, HME87, to treatment with the ultimate carcinogen benzo[a]pyrene diol epoxide (BPDE) were analyzed using a directed gene expression analysis technique, RAGE. Under conditions where cell number was decreased by 50% 24 or 48 h post-treatment, flow cytometry demonstrated no establishment of a G(1)/S arrest nor induction of apoptosis; cells continued to enter S phase from G(1) for at least 24 h but were blocked at G(2)/M. Using the RAGE technique, changes in gene expression were assayed for over 1000 genes, and multiple time-point data were collected for approximately 90 genes. In accord with the cell cycle studies, expression of the p21-WAF1 gene, the major mediator of p53-dependent G(1)/S arrest, did not increase until 24 h post-treatment. The expression of other target genes for transactivation by p53 was increased at early time points, including GADD45, an effector of the G(2)/M checkpoint, and WIP1. Analyses of proteins in treated cells indicated that p53 was phosphorylated at Ser15 but not at Ser20 within 30 min of treatment, and this correlated with an increase in the total amount of p53 protein. Significant expression changes were noted in a number of transcription factor genes, including ATF3 and E2A, genes that have not been previously connected to a response to DNA damage involving bulky chemical adducts. In addition, expression of the XPC gene was induced by BPDE treatment; the XPC product is thought to be involved in recognition of DNA damage by the nucleotide excision repair system. PMID- 12584172 TI - Mutagenicity of reactive oxygen and nitrogen species as detected by co-culture of activated inflammatory leukocytes and AS52 cells. AB - Activated inflammatory leukocytes generate a variety of reactive oxygen and nitrogen species (RONS) that may have roles in mutagenesis and carcinogenesis. The purpose of the present study was to explore the relationship between inflammatory leukocyte activation and mutagenesis using co-culture systems. We investigated the mutagenic potentials of 12-O-tetradecanoylphorbol-13-acetate (TPA)-stimulated differentiated HL-60 (human promyelocytic leukemia cells), and RAW 264.7 cells (murine macrophages) stimulated with lipopolysaccharide (LPS) and interferon (IFN)-gamma by co-culturing each cell line with AS52 cells, a transgenic Chinese hamster ovary cell line. HL-60 cells rapidly generated superoxide (O(2)(-)) 15 min to 1 h (peak at 30 min) following TPA stimulation. RAW 264.7 cells stimulated with LPS and IFN-gamma produced O(2)(-), nitric oxide (NO) and peroxynitrite (ONOO(-)) continuously for 5-25 h. There was a 2.0-fold increase in the mutation frequency of the gpt gene in AS52 cells co-cultured with TPA stimulated HL-60 cells, when compared with non-treated cells. Importantly, this increase in mutation frequency was significantly suppressed by antioxidants, such as superoxide dismutase (SOD) and diphenylene iodonium (DPI), an NADPH oxidase inhibitor (inhibition rates: IRs = 18.2 and 35.1%, respectively). Similarly, co-culture of AS52 cells with LPS/IFN-gamma-stimulated RAW 264.7 cells also increased the mutation frequency of the gpt gene by 2.6-fold, and this increase in mutation frequency was suppressed by SOD, DPI and N(5)-(1-iminoethyl) L-ornithine dihydrochloride (L-NIO), an specific iNOS inhibitor (IRs = 58.3, 70.8 and 70.8%, respectively). In co-culture experiments, activated HL-60 and RAW 264.7 cells increased 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels in AS52 cells when compared with non-treated controls (1.7- and 1.6-fold, respectively). Treatment of AS52 cells with hydrogen peroxide (H(2)O(2), 100 micro M), ONOO(-) (100 micro M) and SIN-1 (100 micro M), a ONOO(-) generator, also increased the mutation frequency of the gpt gene (4.6-, 5.4- and 2.8-fold, respectively). Taken together, these results support the hypothesis that RONS, derived from activated inflammatory leukocytes, are mutagenic in the biological systems, and that RONS generation inhibitors are potentially anti-mutagenic, and thus may be useful in cancer preventive strategies. PMID- 12584174 TI - Reduced apoptotic response to camptothecin in CHO cells deficient in XRCC3. AB - Eukaryotic cells respond to DNA damage by activation of DNA repair, cell-cycle arrest and apoptosis. Several reports suggest that such responses may be coordinated by communication between damage repair proteins and proteins signalling other cellular responses. The Rad51-guided homologous recombination (HR) repair plays an important role in recognition and repair of DNA double strand breaks (DSBs) and cells deficient in this repair pathway become hypersensitive to agents that induce DSBs. In the work reported here we investigated the possible role of the Rad51-like HR proteins XRCC2, XRCC3 and Rad51C in apoptosis following the induction of DSBs by camptothecin. We show that a hamster cell line (irs1SF) deficient in the HR repair gene XRCC3 exhibits altered death and cell-cycle checkpoint responses following treatment with growth inhibitory concentrations of camptothecin. In contrast, hamster cells defective in XRCC2 (irs1) or Rad51C (irs3) treated with equally toxic doses of this agent exhibit a rapid induction of apoptosis similar to that seen in the parental cell line or mutant cells corrected for the HR defect. These results suggest that XRCC3 activity may be necessary for efficient entry into apoptosis in response to DSBs. PMID- 12584173 TI - 15-Lipoxygenase-1 mediates cyclooxygenase-2 inhibitor-induced apoptosis in gastric cancer. AB - It has been found that expression of 15-lipoxygenase-1 (15-LOX-1) and its main product, 13-S-hydroxyoctadecadienoic acid (13-S-HODE), are decreased in human colorectal and esophageal cancers and that non-steroidal anti-inflammatory drugs (NSAIDs) can therapeutically induce 15-LOX-1 expression to trigger apoptosis in those cancer cells. We found that a specific cyclooxygenase-2 (COX-2) inhibitor SC-236 similarly induced apoptosis in gastric cancer cells. In the present study, we tested whether SC-236 induced apoptosis through up-regulation of 15-LOX-1 in gastric cancer. We found that: (i) SC-236 inhibited growth of gastric cancer cells mainly by inducing apoptosis; (ii) SC-236 induced 15-LOX-1 expression and increased endogenous 13-S-HODE product, instead of 15-S-HETE during apoptosis; (iii) SC-236 did not affect expression of COX-1, COX-2, 5-LOX and 12-LOX; and (iv) 15-LOX-1 inhibition suppressed SC-236 induced apoptosis. These findings demonstrated that SC-236 induced apoptosis in gastric cancer cells via up regulation of 15-LOX-1, and 13-S-HODE. These are potential and new targets for prevention and treatment of gastric cancer. PMID- 12584175 TI - Chemoprevention of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ)-induced colonic and hepatic preneoplastic lesions in the F344 rat by cruciferous vegetables administered simultaneously with the carcinogen. AB - The aim of this study was to investigate the chemopreventive effects of widely consumed cruciferous vegetables, namely Brussels sprouts and red cabbage towards 2-amino-3-methylimidazo[4,5-f]quinoline (IQ)-induced preneoplastic lesions [liver glutathione-S-transferase placental positive (GST-P(+)) foci and colonic aberrant crypt foci (ACF)]. Male F344 rats were treated with IQ (100 mg/kg bw/g) on 10 alternating days and received drinking water supplemented with Brussels sprouts and red cabbage juices (5% v/v) before and during the carcinogen treatment. From each vegetable two different cultivars were tested. Brussels sprouts reduced the frequency of IQ-induced aberrant foci in both organs (41-52% in the colon and 27 67% in the liver). Also, Brussels sprouts drastically diminished (85-91%) the size of liver GST-P(+) foci, but no such effect was seen in the colon. With red cabbage, the size of liver GST-P(+) foci was markedly reduced (41-83%) whereas the foci frequency was only moderately decreased (19-50%). No protection was seen in the colon after treatment with red cabbage. Cooking (10 min, 100 degrees C) of the vegetables had no influence on their protective effects. The stronger chemoprotective effects of Brussels sprouts may be due to the fact that the overall glucosinolate contents were substantially (2-3-fold) higher than those of the cabbage cultivars, but it was not possible to attribute the reduction of preneoplastic lesions to specific glucosinolates. The activities of hepatic UDP glucuronosyltransferase form 2 (UDPGT-2) and cytochrome P4501A2 were increased by both vegetables. The induction effect of Brussels sprouts on the activity of UDPGT-2 was more marked than that of the red cabbage cultivars, suggesting that increased glucuronidation of IQ may account for the reduction of the preneoplastic lesions. Our findings support the assumption that Brassica vegetables protect against the carcinogenic effects of heterocyclic amines. PMID- 12584178 TI - Investigation of interaction between N-acetyltransferase 2 and heterocyclic amines as potential risk factors for colorectal cancer. AB - Fast N-acetyltransferase 2 (NAT2) acetylators may be at increased risk of colorectal cancer through the activation of carcinogenic heterocyclic amines (HA), which are produced by meat cooked at high temperatures and are found in cigarette smoke. A study of 500 incident colorectal cancer cases and population controls, matched for age, sex and general practitioner, was conducted in the UK to investigate this hypothesis. Usual meat intake and lifetime smoking habits were estimated using a detailed questionnaire administered by interview. Subjects also indicated how well cooked they ate their meat. Subjects were classified as fast or slow NAT2 acetylators on the basis of NAT2 genotype. Complete genotype data were available on 433 matched pairs. The risk of colorectal cancer showed a steady increase with meat intake, rising to an odds ratio of 1.51 [95% confidence interval (1.03, 2.23)] for the highest versus the lowest quartile, after adjustment for total energy intake, and this was even more pronounced for red meat [odds ratio 1.97 (1.30, 2.98)]. However, this effect was not influenced by the preference for well-done meat. Smoking was also associated with an increased risk [odds ratio 1.47 (1.10, 1.98) for ever- versus never-smokers]. In both cases and controls approximately 40% of subjects were classified as fast acetylators, and the risks associated with (red) meat intake and smoking did not vary with NAT2 status. This study provides no support for the hypothesis that fast NAT2 acetylators are at increased risk of colorectal cancer, even if exposed to high levels of HA from well-cooked meat or smoking. PMID- 12584176 TI - Suppression of tumorigenesis in the Apc(min) mouse: down-regulation of beta catenin signaling by a combination of tea plus sulindac. AB - Epidemiological and animal studies suggest that tea may be protective towards cancers of the GI tract. White tea, the least processed form of tea, contains high levels of polyphenols and, like green tea, is chemopreventive towards heterocyclic amine-initiated colonic aberrant crypt formation in male F344 rats. We examined for the first time the relative effectiveness of white and green tea in suppressing intestinal tumorigenesis in C57BL/6J-Apc(Min/+) (Apc(min)) mice. Each tea was also compared with sulindac, a non-steroidal anti-inflammatory drug known to be highly effective in Apc(min) mice. Male C57BL/6J (+/+) (wild-type) and Apc(min) mice were treated in the drinking water with white tea or green tea (1.5% w/v, 2 min brew-time), 80 p.p.m. sulindac, a combination of 80 p.p.m. sulindac in 1.5% white tea, or pH buffered water. After 12 weeks of treatment, Apc(min) mice given white tea, green tea, or sulindac had significantly fewer tumors than controls (P < 0.05). The protection provided by 1.5% green or white tea was comparable to that provided by 80 p.p.m. sulindac. Mice treated with a combination of white tea plus sulindac had significantly fewer tumors than either treatment alone (P < 0.05). beta-catenin and beta-catenin/Tcf-4 regulated proteins Cyclin D(1) and c-Jun were readily detected in polyps, but markedly reduced in normal-looking intestines of mice treated with both tea and sulindac. This research provides evidence that teas, particularly when administered in combination with sulindac, are highly effective at inhibiting intestinal neoplasia in male Apc(min) mice via direct or indirect effects on the beta catenin/APC pathway. PMID- 12584177 TI - Bleomycin-induced chromosome breaks as a risk marker for lung cancer: a case control study with population and hospital controls. AB - Environmental exposure to carcinogens and individual susceptibility play significant roles in cancer risk. Suboptimal DNA repair capability, measured by quantifying mutagen-induced chromosome breaks, might explain variable host susceptibility to environmental carcinogens. In an ongoing lung cancer case control study, we compared individual sensitivity to bleomycin-induced chromosome breaks in 152 non-small cell lung cancer patients with 94 population controls and 85 hospital controls with no history of cancer. Mutagen sensitivity was measured by mean number of chromatid breaks per cell in cultured peripheral blood lymphocytes treated with bleomycin. Non-parametric tests and chi(2) tests were used to determine the statistical significance of the crude case-control comparisons, followed by logistic regression to adjust for important covariates. The mean number of bleomycin-induced breaks per cell was 1.01 for the cases compared with 0.86 for hospital controls (P < 0.01) and 0.89 for population controls (P < 0.01). The mean number of breaks per cell was 1.01 for those >65 years old and 0.81 for those < or = 65 years old (P < 0.01) among population controls. Defining bleomycin sensitive as >0.84 break/cell (the median level in population controls), 67% of the cases were bleomycin sensitive compared with 49% of the hospital controls [adjusted odds ratio (OR) = 2.69, 95% confidence interval (CI) = 1.44, 5.04], and 51% of the population controls (adjusted OR = 2.18, 95% CI = 1.13, 4.21). Our data indicate that the increased number of bleomycin-induced chromosome breaks was significantly associated with an increased risk of lung cancer in the first 331 subjects. PMID- 12584179 TI - Dietary factors and the occurrence of truncating APC mutations in sporadic colon carcinomas: a Dutch population-based study. AB - The interactions between environmental factors and the genetic and epigenetic changes that drive colon carcinogenesis are not clear. Dietary factors reported previously to be associated with colon cancer risk may well influence the occurrence of specific somatic alterations in colon tumors. To explore this idea, data from a Dutch population-based case-control study (184 cases, 259 controls) on sporadic colon cancer were used to assess associations between dietary factors and the occurrence of truncating mutations in the adenomatous polyposis coli (APC) gene in carcinomas. Single-strand conformation polymorphism analysis and DNA sequencing were used to screen tumors for mutations in the mutation cluster region of APC. Usual dietary habits were assessed by an interview-based questionnaire. Truncating APC mutations were detected in 63 (34%) of the tumors. Vegetable consumption was inversely associated with APC(+) (with mutation) tumors [odds ratio (OR) and 95% confidence interval (CI) for highest versus lowest tertile, OR: 0.6, 95% CI: 0.3-1.3] as well as APC(-) (without mutation) tumors (OR: 0.3, 95% CI: 0.2-0.5). Alcohol intake was positively associated with APC(-) tumors (OR: 1.7, 95% CI: 1.0-3.0) and inversely associated with APC(+) tumors (OR: 0.5, 95% CI: 0.3-1.1). Positive associations were observed for meat, fish and fat with APC(+) tumors (OR: 1.7, 95% CI: 0.8-3.6; OR: 1.4, 95% CI: 0.7-2.8; OR: 4.5, 95% CI: 1.6-12.8, respectively). Of the dietary factors examined, vegetable consumption and alcohol intake were significantly different related to APC(+) tumors than to APC(-) tumors (APC(+) versus APC(-), OR: 2.3, 95% CI: 1.0 5.3; OR: 0.3, 95% CI: 0.2-0.7, respectively). Our data suggest that vegetables play a protective role in the etiology of both tumor subsets, although this role appears to be less influential in the APC(+) group. Alcohol seems to especially promote the development of APC(-) tumors whereas meat, fish and fat appear to enhance the development of APC(+) tumors. PMID- 12584180 TI - Comparative metabolism of N-nitrosopiperidine and N-nitrosopyrrolidine by rat liver and esophageal microsomes and cytochrome P450 2A3. AB - N-nitrosopiperidine (NPIP) is a potent esophageal carcinogen in rats whereas structurally similar N-nitrosopyrrolidine (NPYR) induces liver, but not esophageal tumors. NPIP is a possible causative agent for human esophageal cancer. Our goal is to explain mechanistically these differing carcinogenic activities in the esophagus. We hypothesize that differences in metabolic activation of these nitrosamines could be one factor accounting for their differing carcinogenicity. alpha-Hydroxylation is the key metabolic activation pathway leading to nitrosamine-induced carcinogenesis. In this study, we examined the alpha-hydroxylation rates of [3,4-(3)H]NPIP and [3,4-(3)H]NPYR by male F344 rat esophageal and liver microsomes. The major alpha-hydroxylation products of NPIP and NPYR, 2-hydroxytetrahydro-2H-pyran (2-OH-THP) and 2 hydroxytetrahydrofuran (2-OH-THF), respectively, were monitored by high performance liquid chromatography with radioflow detection. NPIP or NPYR (4 microM) was incubated with varying concentrations of esophageal microsomes and co factors. Microsomes converted NPIP to 2-OH-THP with a 40-fold higher velocity than NPYR to 2-OH-THF. Similar results were observed in studies with NPIP and NPYR at substrate concentrations between 4 and 100 micro M. Kinetics of NPIP alpha-hydroxylation were biphasic; K(M) values were 312 +/- 50 and 1600 +/- 312 microM. Expressed cytochrome P450 2A3, found in low levels in rat esophagus, was a good catalyst of NPIP alpha-hydroxylation (K(M) = 61.6 +/- 20.5 microM), but a poor catalyst of NPYR alpha-hydroxylation (K(m) = 1198 +/- 308 micro M). Cytochrome P450 2A3 may play a role in the preferential activation of NPIP observed in rat esophagus. Liver microsomes metabolized NPYR to 2-OH-THF (V(max)/K(M) = 3.23 pmol/min/mg/ microM) as efficiently as NPIP to 2-OH-THP (V(max)/K(M) = 3.80-4.61 pmol/min/mg/ microM). We conclude that rat esophageal microsomes activate NPIP but not NPYR whereas rat liver microsomes activate NPIP and NPYR. These results are consistent with previous findings that tissue specific activation of nitrosamines contributes to tissue-specific tumor formation. PMID- 12584181 TI - Endogenous retroviral elements, but not exogenous retroviruses, are detected in CD30-positive lymphoproliferative disorders of the skin. PMID- 12584182 TI - Inverse association between phospholipase A2 and COX-2 expression during mouse colon tumorigenesis. AB - Cytosolic phospholipase A(2) (cPLA(2)) releases arachidonic acid (AA) from intracellular phospholipids. We evaluated the status of cPLA(2) in azoxymethane (AOM)-induced mouse colon tumors. Despite increased expression of cyclooxygenase 2 (3.7-fold) and PGE(2) (3.4-fold) production in tumors, cPLA(2) mRNA levels and enzyme activity were significantly reduced (3.6- and 3-fold, respectively). Reduced levels of cPLA(2) were also observed in pre-neoplastic aberrant crypt foci (ACF), a distinct morphological alteration that represents an early stage in the pathogenesis of colon tumors. Furthermore, the reciprocal expression patterns of these two genes were found to occur in human colorectal cancers (CRC). Examination of the activity of the secretory phospholipases A(2) (sPLA(2)) and expression of the groups V and X sPLA(2)s revealed no compensatory increase in tumor tissue. As cPLA(2) has been shown to be involved in TNF-alpha-induced apoptosis in certain cell types, and TNF-alpha expression is significantly enhanced in AOM-induced tumors (2.8-fold), we examined the role of cPLA(2) in TNF alpha-induced apoptosis of cultured mouse colonocytes (YAMC). The specific cPLA(2) inhibitor, AACOCF(3) (arachidonoyl trifluoromethyl ketone), was able to protect colonocytes from TNF-alpha-induced apoptosis in vitro. In summary, our data demonstrate an inverse relationship between COX-2 and cPLA(2) expression in both AOM-induced mouse colon tumors and human CRC and suggest that down regulation of cPLA(2) may attenuate TNF-alpha mediated apoptosis during tumorigenesis and facilitate tumor progression. PMID- 12584183 TI - PKC isozyme S-cysteinylation by cystine stimulates the pro-apoptotic isozyme PKC delta and inactivates the oncogenic isozyme PKC epsilon. AB - Protein kinase C (PKC) is a family of ten isozymes that play distinct and in some cases opposing roles in cell growth and survival. We recently reported that diamide, a diazene carbonyl derivative which oxidizes thiols to disulfides through addition/displacement reactions at the diazene bond, induces potent GSH dependent inactivation of several PKC isozymes, including the oncogenic isozyme PKC epsilon, via S-glutathiolation. PKC delta, a pro-apoptotic isozyme, was distinguished by its resistance to inactivation. In this report, we show that PKC regulatory S-thiolation modifications produced by physiological disulfides elicit opposing effects on PKC delta and PKC epsilon activity. We report that PKC delta is stimulated 2.0-2.5 fold by GSSG, (Cys-Gly)(2) and cystine, under conditions where PKC gamma and PKC epsilon are fully inactivated by cystine, and PKC alpha activity is affected marginally or not at all by the disulfides. Focusing on cystine, we show that DTT quenches cystine-induced PKC delta stimulation and PKC gamma and PKC epsilon inactivation, indicative of oxidative regulation. By analyzing DTT-reversible isozyme radiolabeling by [(35)S]cystine, we demonstrate that PKC gamma, PKC delta and PKC epsilon are each [(35)S] S-cysteinylated in association with the concentration-dependent regulation of isozyme activity by cystine. The restricted reactivity of cystine, together with the effects of DTT and thioredoxin on cystine-induced PKC isozyme regulation reported here, indicate that the cystine-induced PKC-regulatory effects entail isozyme S-cysteinylation. We recently hypothesized that antagonism of tumor promotion/progression by small cellular thiols may involve PKC regulation via oxidant-induced S-thiolation reactions with PKC isozymes. The findings of cystine-induced PKC isozyme regulation by S-cysteinylation reported here offer correlative support to the hypothetical model. Thus, PKC delta, a potent antagonist of DMBA-TPA-induced tumor promotion/progression in mouse skin, is stimulated by S-cysteinylation, PKC epsilon, an important mediator of the tumor promotion/progression response, is inactivated by S-cysteinylation, and PKC alpha, which is not influential in DMBA TPA-induced tumor promotion/progression, is not regulated by cystine. Furthermore, PKC gamma has oncogenic activity, and S-cysteinylation inactivated PKC gamma and PKC epsilon similarly. These findings provide evidence that S cysteinyl acceptor-sites in PKC isozymes may offer attractive targets for development of novel cancer preventive agents. PMID- 12584184 TI - CYP1B1 determines susceptibility to low doses of 7,12-dimethylbenz[a]anthracene induced ovarian cancers in mice: correlation of CYP1B1-mediated DNA adducts with carcinogenicity. AB - We showed previously that CYP1B1-null mice developed 10 times less lymphomas than wild-type mice after receiving 7,12-dimethylbenz[a]anthracene (DMBA). In this study a 10-fold lower dose was applied to differentiate between toxicity induced lymphomas (200 micro g/mouse/day) and tumor initiation (20 micro g/day). DMBA adducts to DNA of organs of mice, or to DNA of V79 cells expressing single mice or human cytochrome P450 isoenzymes were also measured. Mice were dosed three cycles of 5 days/week with DMBA in corn oil orally. Histopathology was determined at intermittent death or 1 year after dosing. DMBA-DNA adducts were assayed by (32)P-postlabeling. At 20 micro g/day, wild-type mice developed ovary (71%, stromal cells derived), skin (36%), uterus (64%) and lung (14%) hyperplasias. At this dose the CYP1B1-null mice developed no lymphomas, 25% ovary (epithelial cells derived), 8% skin, 58% uterus and 33% lung tumors. Oil control mice (n = 35) developed only eight, mostly different, hyperplasias. Wild-type mice had more DMBA-DNA adducts than the CYP1B1-null mice. The differences were highest in thymus, spleen, ovaries and testes (5-7-fold). Additionally, one specific DMBA DNA adduct was reduced in CYP1B1-null mice. V79-cells expressed mouse CYP1B1 was 35 times more active than mouse CYP1A1 in forming DMBA-DNA adducts. Human CYP1B1 was 2.5 times less active than mouse CYP1B1 but 2.3-fold more active than human CYP1A1. CYP1B1 is the dominant enzyme in metabolizing DMBA to carcinogenic metabolites at high and low doses in mice, leading to an increased tumor rate of especially the ovaries at low doses of DMBA. Wild-type mice had more DMBA-DNA adducts than CYP1B1-null mice. Additionally, a specific adduct was less present in the CYP1B1-null mice. Human CYP1B1 was less active than mouse CYP1B1, but more active than human CYP1A1 in forming DMBA-DNA adducts. Thus, we expect CYP1B1 to be an important DMBA activating enzyme in humans also. PMID- 12584185 TI - Carcinogenicity of dimethylarsinic acid in p53 heterozygous knockout and wild type C57BL/6J mice. AB - There is abundant epidemiological evidence that arsenic is an environmental carcinogen related to human cancers of the skin, lung, liver and urinary bladder, in particular. Dimethylarsinic acid (DMA) has also been reported to act as a carcinogen/or a promoter in rat models. To elucidate molecular mechanisms, we conducted an 18 month carcinogenicity study of DMA in p53 heterozygous (+/-) knockout mice, which are susceptible to early spontaneous development of various types of tumors, and wild-type (+/+) C57BL/6J mice. Totals of 88-90 males, 7-8 weeks of age, were divided into three groups each administered 0, 50 or 200 p.p.m. DMA in their drinking water for 18 months. Mice that were found moribund or died before the end of the study were autopsied to evaluate the tumor induction levels, as well as those killed at the end. Both p53(+/-) knockout and wild-type mice demonstrated spontaneous tumor development, but lesions were more prevalent in the knockout case. Carcinogenic effect of DMA was evident by significant early induction of tumors in both treated p53(+/-) knockout and wild type mice, significant increase of the tumor multiplicity in 200 p.p.m.-treated p53(+/-) knockout mice, and by significant increase in the incidence and multiplicity of tumors (malignant lymphomas) in the treated wild-type mice. By the end of 80 weeks, tumor induction, particularly malignant lymphomas and sarcomas, were similar in treated and control p53(+/-) knockout mice. No evidence for organ-tumor specificity of DMA was obtained. Molecular analysis using PCR SSCP techniques revealed no p53 mutations in lymphomas from either p53(+/-) knockout or wild-type mice. In conclusion, DMA primarily exerted its carcinogenic effect on spontaneous development of tumors with both of the animal genotypes investigated here. PMID- 12584186 TI - The trophic effect of dietary fibre is not associated with a change in total crypt number in the distal colon of rats. AB - Soluble fibres, such as guar gum, promote and wheat bran or methylcellulose protect from chemically induced colon carcinogenesis, relative to the effect of a fibre-free diet in rats. Mechanisms are poorly understood. Whereas all fibres are trophic to the colonic epithelium, the heterogeneity of effects on carcinogenesis may reflect different effects on the total number of crypts and, therefore, the size of the stem cell population. This study aimed to assess this hypothesis. Sprague-Dawley rats were fed one of fibre-free diets with or without 10% wheat bran, methylcellulose or guar gum for 4 weeks. The distal colons were stained with methylene blue and quantified for the number and density of crypts using an image analysis system. Epithelial proliferative kinetics was measured stathmokinetically. Methodology for quantifying crypts was valid and reproducible. Rats fed a fibre-free diet had atrophic distal colon, as shown by a decrease in crypt column height and a lower mitotic index. Fibre supplementation prevented the atrophy and was associated with crypt mouth areas that were 30-60% larger than those in the fibre-free group (P < 0.001, ANOVA), with the methylcellulose group being the largest (1.16 microm(2)). The crypt density of the fibre-free group was 16-19% greater than those in fibre fed groups (P + 0.006), due to the smaller size of the crypts. However, there was no difference in the total number of crypts across the four dietary groups (P > 0.1). Distal colons in all of the dietary groups contained approximately 10(5) crypts. In conclusion, although variation in the amount or type of dietary fibre exerts heterogeneous effects on the growth of the colonic epithelium and on colon carcinogenesis, the total number of crypts in the distal colon remains constant. It is, therefore, unlikely that fibres influence carcinogenic events by altering the size of the stem cell population. PMID- 12584187 TI - Acid-sensing ion channels in malignant gliomas. AB - High grade glioma cells derived from patient biopsies express an amiloride sensitive sodium conductance that has properties attributed to the human brain sodium channel family, also known as acid-sensing ion channels (ASICs). This amiloride-sensitive conductance was not detected in cells obtained from normal brain tissue or low grade or benign tumors. Differential gene profiling data showed that ASIC1 and ASIC2 mRNA were present in normal and low grade tumor cells. Although ASIC1 was present in all of the high grade glial cells examined, ASIC2 mRNA was detected in less than half. The main purpose of our work was to examine the molecular mechanisms that may underlie the constitutively activated sodium currents present in high grade glioma cells. Our results show that 1) gain of-function mutations of ASIC1 were not present in a number of freshly resected and cultured high grade gliomas, 2) syntaxin 1A inhibited ASIC currents only when ASIC1 and ASIC2 were co-expressed, and 3) the inhibition of ASIC currents by syntaxin 1A had an absolute requirement for either gamma- or delta-hENaC. Transfection of cultured cells originally derived from high grade gliomas (U87-MG and SK-MG1) with ASIC2 abolished basal amiloride-sensitive sodium conductance; this inhibition was reversed by dialysis of the cell interior with Munc-18, a syntaxin-binding protein that typically blocks the interaction of syntaxin with other proteins. Thus, syntaxin 1A cannot inhibit Na(+) permeability in the absence of adequate plasma membrane ASIC2 expression, accounting for the observed functional expression of amiloride-sensitive currents in high grade glioma cells. PMID- 12584188 TI - Inactivation of retinoblastoma (RB) tumor suppressor by oncogenic isoforms of the p53 family member p73. AB - The p53 family includes three members that share significant sequence homology, yet exhibit fundamentally different functions in tumorigenesis. Whereas p53 displays all characteristics of a classical tumor suppressor, its homologues p63 and p73 do not. We have previously shown, that NH(2)-terminally truncated isoforms of p73 (Delta TA-p73), which act as dominant-negative inhibitors of p53 are frequently overexpressed in cancer cells. Here we provide evidence that Delta TA-p73 isoforms also affect the retinoblastoma protein (RB) tumor suppressor pathway independent of p53. Delta TA-p73 isoforms inactivate RB by increased phosphorylation, resulting in enhanced E2F activity and proliferation of fibroblasts. By inactivating the two major tumor suppressor pathways in human cells they act functionally analogous to several viral oncoproteins. These findings provide an explanation for the fundamentally different functions of p53 and p73 in tumorigenesis. PMID- 12584189 TI - Glycogen synthase kinase 3 beta is a natural activator of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 1 (MEKK1). AB - Glycogen synthase kinase 3beta (GSK3 beta) is implicated in many biological events, including embryonic development, cell differentiation, apoptosis, and insulin response. GSK3 beta has now been shown to induce activation of the mitogen-activated protein kinase kinase kinase MEKK1 and thereby to promote signaling by the stress-activated protein kinase pathway. GSK3 beta-binding protein blocked the activation of MEKK1 by GSK3 beta in human embryonic kidney 293 (HEK293) cells. Furthermore, co-immunoprecipitation analysis revealed a physical association between endogenous GSK3 beta and MEKK1 in HEK293 cells. Overexpression of axin1, a GSK3 beta-regulated scaffolding protein, did not affect the physical interaction between GSK3 beta and MEKK1 in transfected HEK293 cells. Exposure of cells to insulin inhibited the activation of MEKK1 by GSK3 beta, and this inhibitory effect of insulin was abolished by the phosphatidylinositol 3-kinase inhibitor wortmannin. Furthermore, MEKK1 activity under either basal or UV- or tumor necrosis factor alpha-stimulated conditions was reduced in embryonic fibroblasts derived from GSK3 beta knockout mice compared with that in such cells from wild-type mice. Ectopic expression of GSK3 beta increased both basal and tumor necrosis factor alpha-stimulated activities of MEKK1 in GSK3 beta(-/-) cells. Together, these observations suggest that GSK3 beta functions as a natural activator of MEKK1. PMID- 12584190 TI - Mammalian translesion DNA synthesis across an acrolein-derived deoxyguanosine adduct. Participation of DNA polymerase eta in error-prone synthesis in human cells. AB - alpha-OH-PdG, an acrolein-derived deoxyguanosine adduct, inhibits DNA synthesis and miscodes significantly in human cells. To probe the cellular mechanism underlying the error-free and error-prone translesion DNA syntheses, in vitro primer extension experiments using purified DNA polymerases and site-specific alpha-OH-PdG were conducted. The results suggest the involvement of pol eta in the cellular error-prone translesion synthesis. Experiments with xeroderma pigmentosum variant cells, which lack pol eta, confirmed this hypothesis. The in vitro results also suggested the involvement of pol iota and/or REV1 in inserting correct dCMP opposite alpha-OH-PdG during error-free synthesis. However, none of translesion-specialized DNA polymerases catalyzed significant extension from a dC terminus when paired opposite alpha-OH-PdG. Thus, our results indicate the following. (i) Multiple DNA polymerases are involved in the bypass of alpha-OH PdG in human cells. (ii) The accurate and inaccurate syntheses are catalyzed by different polymerases. (iii) A modification of the current eukaryotic bypass model is necessary to account for the accurate bypass synthesis in human cells. PMID- 12584191 TI - Biological processing of the cocaine and amphetamine-regulated transcript precursors by prohormone convertases, PC2 and PC1/3. AB - Cocaine and amphetamine-regulated transcript (CART), a neuroendocrine peptide influencing reward, feeding/appetite, and stress responses is derived from two peptide precursors of 129 and 116 amino acid (aa) residues that arise via alternative splicing from a single Cart gene in rats and mice. The signal peptide constitutes the first 27 aa resulting in pro-CART molecules of either 102 or 89 aa. In the present study, we have shown that pro-CART is a substrate for the neuroendocrine subtilisin/kexin-like prohormone convertases, PC2 (SPC2) and PC1/3 (SPC3). By using different neuroendocrine cell lines, with or without endogenous expression of either PC2 or PC1/3 or both enzymes, we have demonstrated through transient transfection studies that long pro-CART gives rise to an intermediate peptide, residues 33-102, and the two major bioactive CART forms, residues 55-102 (I) and 62-102 (II), respectively. Likewise, short pro-CART also generates three peptides, an intermediate, residues 10-89, and the two identical bioactive CART forms. We have confirmed the identities of the bioactive and intermediate CART molecules by microsequencing and/or high performance liquid chromatography and mass spectrometry. We have shown that PC2 is more efficient in generating bioactive CART I compared with PC1/3, whereas the production of the smaller bioactive CART II is exclusively carried out by PC2. PC1/3 is predominantly responsible for generating the intermediate CART fragments, 33-102 and 10-89, from long and short pro-CART, respectively. To compare in vitro and in vivo processing of pro-CART, we have examined its processing in PC2, 7B2, and PC1/3 knock-out mouse hypothalamic extracts and demonstrated that, as in vitro, PC2 is more potent than PC1/3 in generating bioactive CART I whereas bioactive CART II is solely generated by PC2. Also, in vivo, we have shown that PC1/3 is predominantly active in liberating the two intermediate CART fragments, 33-102 and 10-89. These findings confirm the key roles of PC2 and PC1/3 acting together or separately to carry out CART processing in selected sites in vivo. PMID- 12584192 TI - Fes tyrosine kinase promotes survival and terminal granulocyte differentiation of factor-dependent myeloid progenitors (32D) and activates lineage-specific transcription factors. AB - The c-fps/fes proto-oncogene encodes a 92-kDa protein-tyrosine kinase that is involved in myeloid cell development and function. We have recently shown that expression of an activated allele of Fes (Fes(act)) in monocyte precursors resulted in their differentiation into functional macrophages through the activation of lineage-specific transcription factors. We now report that this kinase also plays a role in the survival and terminal differentiation of granulocyte progenitors. The expression of Fes(act) in factor-dependent 32D cells prevented their apoptotic death after interleukin-3 removal, but Fes(act) expressing cells remained factor-dependent for proliferation. Removal of interleukin-3 from the Fes(act)-expressing cells was followed by granulocytic differentiation in the absence of granulocyte colony-stimulating factor within 4 8 days. The differentiated cells had distinctive granulocyte morphology and there was up-regulation of CD11b, Gr-1, and late differentiation markers such as lactoferrin, suggesting that this kinase induced terminal granulocytic differentiation. Concomitantly, Fes(act) down-regulated the macrophage marker F4/80, suggesting that the biological activity of Fes was coordinated in a lineage-specific manner. Further analysis showed that Fes(act) caused activation of CCAAT/enhancer-binding protein-alpha and STAT3, two transcription factors that are involved in granulocyte differentiation. Our results provide evidence that Fes may be a key component of the granulocyte differentiation machinery, and suggest a potential mechanism by which this kinase may regulate granulocyte specific gene expression. PMID- 12584193 TI - Heat shock protein 90 function is essential for Plasmodium falciparum growth in human erythrocytes. AB - Hsp90 is important for normal growth and development in eukaryotes. Together with Hsp70 and other accessory proteins, Hsp90 not only helps newly synthesized proteins to fold but also regulates activities of transcription factors and protein kinases. Although the gene coding for heat shock protein 90 from Plasmodium falciparum (PfHsp90) has been characterized previously, there is very little known regarding its function in the parasite. We have analyzed PfHsp90 complexes and addressed its role in parasite life cycle using Geldanamycin (GA), a drug known to interfere with Hsp90 function. Sedimentation analysis and size exclusion chromatography showed PfHsp90 to be in 11 s(20,(w)) complexes of approximately 300 kDa in size. Similar to the hetero-oligomeric complexes of Hsp90 in mammals, PfHsp70 was found to be present in PfHsp90 complexes. Homology modeling revealed a putative GA-binding pocket at the amino terminus of PfHsp90. The addition of GA inhibited parasite growth with LD(50) of 0.2 microm. GA inhibited parasite growth by arresting transition from Ring to trophozoite. Transition from trophozoite to schizonts and reinvasion of new erythrocytes were less significantly affected. While inducing the synthesis of PfHsp70 and PfHsp90, GA did not significantly alter the pattern of newly synthesized proteins. Pre exposure to heat shock attenuated GA-mediated growth inhibition, suggesting the involvement of heat shock proteins. Specificity of GA action on PfHsp90 was evident from selective inhibition of PfHsp90 phosphorylation in GA-treated cultures. In addition to suggesting an essential role for PfHsp90 during parasite growth, our results highlight PfHsp90 as a potential drug target to control malaria. PMID- 12584194 TI - The ALD6 gene product is indispensable for providing NADPH in yeast cells lacking glucose-6-phosphate dehydrogenase activity. AB - Reducing equivalents in the form of NADPH are essential for many enzymatic steps involved in the biosynthesis of cellular macromolecules. An adequate level of NADPH is also required to protect cells against oxidative stress. The major enzymatic source of NADPH in the cell is the reaction catalyzed by glucose-6 phosphate dehydrogenase, the first enzyme in the pentose phosphate pathway. Disruption of the ZWF1 gene, encoding glucose-6-phosphate dehydrogenase in the yeast Saccharomyces cerevisiae, results in methionine auxotrophy and increased sensitivity to oxidizing agents. It is assumed that both phenotypes are due to an NADPH deficiency in the zwf1Delta strain. We used a Met(-) phenotype displayed by the zwf1Delta strain to look for multicopy suppressors of this deletion. We found that overexpression of the ALD6 gene coding for cytosolic acetaldehyde dehydrogenase, which utilizes NADP(+) as its cofactor, restores the Met(+) phenotype of the zwf1Delta strain. Another multicopy suppressor identified in our screen, the ZMS1 gene encoding a putative transcription factor, regulates the level of ALD6 expression. A strain bearing a double ZWF1 ALD6 gene disruption is not viable. Thus, our results indicate the reaction catalyzed by Ald6p as an important source of reducing equivalents in the yeast cells. PMID- 12584195 TI - Active lipoprotein precursors in the Gram-positive eubacterium Lactococcus lactis. AB - Lipid-modified proteins play important roles at the interface between eubacterial cells and their environment. The importance of lipoprotein processing by signal peptidase II (SPase II) is underscored by the fact that this enzyme is essential for viability of the Gram-negative eubacterium Escherichia coli. In contrast, SPase II is not essential for growth and viability of the Gram-positive eubacterium Bacillus subtilis. This could be due to alternative amino-terminal lipoprotein processing, which was shown previously to occur in SPase II mutants of B. subtilis. Alternatively, uncleaved lipoprotein precursors might be functional. To explore further the importance of lipoprotein processing in Gram positive eubacteria, an SPase II mutant strain of Lactococcus lactis was constructed. Although some of the 39 (predicted) lactococcal lipoproteins, such as PrtM and OppA, are essential for growth in milk, the growth of SPase II mutant L. lactis cells in this medium was not affected. Furthermore, the activity of the strictly PrtM-dependent extracellular protease PrtP, which is required for casein degradation, was not impaired in the absence of SPase II. Importantly, no alternative processing of pre-PrtM and pre-OppA was observed in cells lacking SPase II. Taken together, these findings show for the first time that authentic lipoprotein precursors retain biological activity. PMID- 12584196 TI - Recycling of apoprotein E is associated with cholesterol efflux and high density lipoprotein internalization. AB - After receptor-mediated endocytosis of triglyceride-rich lipoproteins (TRL) into the liver, TRL particles are immediately disintegrated in peripheral endosomal compartments. Whereas core lipids and apoprotein B are delivered for degradation into lysosomes, TRL-derived apoE is efficiently recycled back to the plasma membrane. This is followed by apoE re-secretion and association of apoE with high density lipoproteins (HDL). Because HDL and apoE can independently promote cholesterol efflux, we investigated whether recycling of TRL-derived apoE in human hepatoma cells and fibroblasts could be linked to intracellular cholesterol transport. In this study we demonstrate that HDL(3) does not only act as an extracellular acceptor for recycled apoE but also stimulates the recycling of internalized TRL-derived apoE. Furthermore, radioactive pulse-chase experiments indicate that apoE recycling is accompanied by cholesterol efflux. Confocal imaging reveals co-localization of apoE and cholesterol in early endosome antigen 1-positive endosomes. During apoE re-secretion, HDL(3)-derived apoA-I is found in these early endosome antigen 1, cholesterol-containing endosomes. As shown by time-lapse fluorescence microscopy, apoE recycling involves the intracellular trafficking of apoA-I to pre-existing and TRL-derived apoE/cholesterol-containing endosomes in the periphery. Thus, these studies provide evidence for a new intracellular link between TRL-derived apoE, cellular cholesterol transport, and HDL metabolism. PMID- 12584198 TI - The Caenorhabditis elegans genes sqv-2 and sqv-6, which are required for vulval morphogenesis, encode glycosaminoglycan galactosyltransferase II and xylosyltransferase. AB - In mutants defective in any of eight Caenorhabditis elegans sqv (squashed vulva) genes, the vulval extracellular space fails to expand during vulval morphogenesis. Strong sqv mutations result in maternal-effect lethality, caused in part by the failure of the progeny of homozygous mutants to initiate cytokinesis and associated with the failure to form an extracellular space between the egg and the eggshell. Recent studies have implicated glycosaminoglycans in these processes. Here we report the cloning and characterization of sqv-2 and sqv-6. sqv-6 encodes a protein similar to human xylosyltransferases. Transfection of sqv-6 restored xylosyltransferase activity to and rescued the glycosaminoglycan biosynthesis defect of a xylosyltransferase mutant hamster cell line. sqv-2 encodes a protein similar to human galactosyltransferase II. A recombinant SQV-2 fusion protein had galactosyltransferase II activity with substrate specificity similar to that of human galactosyltransferase II. We conclude that C. elegans SQV-6 and SQV-2 likely act in concert with other SQV proteins to catalyze the stepwise formation of the proteoglycan core protein linkage tetrasaccharide GlcAbeta1,3Galbeta1, 3Galbeta1,4Xylbeta-O-(Ser), which is common to the two major types of glycosaminoglycans in vertebrates, chondroitin and heparan sulfate. Our results strongly support a model in which C. elegans vulval morphogenesis and zygotic cytokinesis depend on the expression of glycosaminoglycans. PMID- 12584199 TI - Cholesterol distribution in the Golgi complex of DITNC1 astrocytes is differentially altered by fresh and aged amyloid beta-peptide-(1-42). AB - The Golgi complex plays an important role in cholesterol trafficking in cells, and amyloid beta-peptides (Abetas) alter cholesterol trafficking. The hypothesis was tested that fresh and aged Abeta-(1-42) would differentially modify Golgi cholesterol content in DINTC1 astrocytes and that the effects of Abeta-(1-42) would be associated with the region of the Golgi complex. Two different methods were used to determine the effects of Abeta-(1-42) on Golgi complex cholesterol. Confocal microscopy showed that fresh Abeta-(1-42) significantly increased cholesterol and that aged Abeta-(1-42) significantly reduced cholesterol content in the Golgi complex. Isolation of the Golgi complex into two fractions using density gradient centrifugation showed effects of aged Abeta-(1-42) similar to those observed with confocal microscopy but revealed the novel finding that fresh Abeta-(1-42) had opposite effects on the two Golgi fractions suggesting a specificity of Abeta-(1-42) perturbation of the Golgi complex. Phosphatidylcholine-phospholipase D activity, cell membrane cholesterol, and apolipoprotein E levels were associated with effects of fresh Abeta-(1-42) on cholesterol distribution but not with effects of aged Abeta-(1-42), arguing against a common mechanism. Extracellular Abeta-(1-42) targets the Golgi complex and disrupts cell cholesterol homeostasis, and this action of Abeta-(1-42) could alter cell functions requiring optimal levels of cholesterol. PMID- 12584197 TI - Identification of mammalian Mediator subunits with similarities to yeast Mediator subunits Srb5, Srb6, Med11, and Rox3. AB - The Mediator is a multiprotein coactivator required for activation of RNA polymerase II transcription by DNA binding transactivators. We recently identified a mammalian homologue of yeast Mediator subunit Med8 and partially purified a Med8-containing Mediator complex from rat liver nuclei (Brower, C. S., Sato, S., Tomomori-Sato, C., Kamura, T., Pause, A., Stearman, R., Klausner, R. D., Malik, S., Lane, W. S., Sorokina, I., Roeder, R. G., Conaway, J. W., and Conaway, R. C. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 10353-10358). Analysis of proteins present in the most highly purified Med8-containing fractions by tandem mass spectrometry led to the identification of many known mammalian Mediator subunits, as well as four potential Mediator subunits exhibiting sequence similarity to yeast Mediator subunits Srb5, Srb6, Med11, and Rox3. Here we present direct biochemical evidence that these four proteins are bona fide mammalian Mediator subunits. In addition, we identify direct pairwise binding partners of these proteins among the known mammalian Mediator subunits. Taken together, our findings identify a collection of novel mammalian Mediator subunits and shed new light on the underlying architecture of the mammalian Mediator complex. PMID- 12584200 TI - Identification of proteases involved in the proteolysis of vascular endothelium cadherin during neutrophil transmigration. AB - Transmigration of neutrophils across the endothelium occurs at the cell-cell junctions where the vascular endothelium cadherin (VE cadherin) is expressed. This adhesive receptor was previously demonstrated to be involved in the maintenance of endothelium integrity. We propose that neutrophil transmigration across the vascular endothelium goes in parallel with cleavage of VE cadherin by elastase and cathepsin G present on the surface of neutrophils. This hypothesis is supported by the following lines of evidence. 1) Proteolytic fragments of VE cadherin are released into the culture medium upon adhesion of neutrophils to endothelial cell monolayers; 2) conditioned culture medium, obtained after neutrophil adhesion to endothelial monolayers, cleaves the recombinantly expressed VE cadherin extracellular domain; 3) these cleavages are inhibited by inhibitors of elastase; 4) VE cadherin fragments produced by conditioned culture medium or by exogenously added elastase are identical as shown by N-terminal sequencing and mass spectrometry analysis; 5) both elastase- and cathepsin G specific VE cadherin cleavage patterns are produced upon incubation with tumor necrosis factor alpha-stimulated and fixed neutrophils; 6) transendothelial permeability increases in vitro upon addition of either elastase or cathepsin G; and 7) neutrophil transmigration is reduced in vitro in the presence of elastase and cathepsin G inhibitors. Our results suggest that cleavage of VE cadherin by neutrophil surface-bound proteases induces formation of gaps through which neutrophils transmigrate. PMID- 12584201 TI - Amyloid-like fibril formation in an all beta-barrel protein. Partially structured intermediate state(s) is a precursor for fibril formation. AB - Acidic fibroblast growth factor from newt (Notopthalmus viridescens) is a approximately 15-kDa, all beta-sheet protein devoid of disulfide bonds. In the present study, we investigate the effects of 2,2,2-trifluoroethanol (TFE) on the structure of newt acidic fibroblast growth factor (nFGF-1). The protein aggregates maximally in 10% (v/v) TFE. Congo red and thioflavin T binding experiments suggest that the aggregates induced by TFE have properties resembling the amyloid fibrils. Transmission electron microscopy and x-ray fiber diffraction data show that the fibrils (induced by TFE) are straight, unbranched, and have a cross-beta structure with an average diameter of 10-15 A. Preformed fibrils (induced by TFE) of nFGF-1 are observed to seed amyloid-like fibril formation in solutions containing the protein (nFGF-1) in the native beta-barrel conformation. Fluorescence, far-UV CD, anilino-8-napthalene sulfonate binding, multidimensional NMR, and Fourier transformed infrared spectroscopy data reveal that formation of a partially structured intermediate state(s) precedes the onset of the fibrillation process. The native beta-barrel structure of nFGF-1 appears to be disrupted in the partially structured intermediate state(s). The protein in the partially structured intermediate state(s) is found to be "sticky" with a solvent exposed non-polar surface(s). Amyloid fibril formation appears to occur due to coalescence of the protein in the partially structured intermediate state(s) through solvent-exposed non-polar surfaces and intermolecular beta-sheet formation among the extended, linear beta-strands in the protein. PMID- 12584202 TI - Oncogenic Ha-Ras transformation modulates the transcription of the CTP:phosphocholine cytidylyltransferase alpha gene via p42/44MAPK and transcription factor Sp3. AB - We have shown previously that expression of the murine CTP:phosphocholine cytidylyltransferase (CT) alpha gene is regulated during cell proliferation (Golfman, L. S., Bakovic, M., and Vance, D. E. (2001) J. Biol. Chem. 276, 43688 43692). We have now characterized the role of Ha-Ras in the transcriptional regulation of the CTalpha gene. The expression of CTalpha and CTbeta2 proteins and mRNAs was stimulated in C3H10T1/2 murine fibroblasts expressing oncogenic Ha Ras. Incubation of cells with the specific inhibitor (PD98059) of p42/44(MAPK) decreased the expression of both CT isoforms. Transfection of fibroblasts with CTalpha promoter-luciferase constructs resulted in an approximately 2-fold enhanced luciferase expression in Ha-Ras-transformed, compared with nontransformed, fibroblasts. Electromobility shift assays indicated enhanced binding of the Sp3 transcription factor to the CTalpha promoter in Ha-Ras transformed cells. Expression of several forms of Sp3 was increased in nuclear extracts of Ha-Ras-transformed fibroblasts compared with nontransformed cells. Tyrosine phosphorylation of one Sp3 form was decreased, whereas phosphorylation of two other forms of Sp3 was increased in nuclear extracts of Ha-Ras-transformed cells. When control fibroblasts were transfected with a Sp3-expressing plasmid, an enhanced expression of CTalpha and CTbeta was observed. However, the expression of CTalpha or CTbeta was not increased in Ha-Ras-transformed cells transfected with a Sp3 plasmid presumably because expression was already maximally enhanced. The results suggest that Sp3 is a downstream effector of a Ras/p42/44(MAPK) signaling pathway which increases CTalpha gene transcription. PMID- 12584203 TI - Store-operated Ca2+ current in prostate cancer epithelial cells. Role of endogenous Ca2+ transporter type 1. AB - Ca(2+) influx via store-operated channels (SOCs) following stimulation of the plasma membrane receptors is the key event controlling numerous processes in nonexcitable cells. The human transient receptor potential vanilloid type 6 channel, originally termed Ca(2+) transporter type 1 (CaT1) protein, is one of the promising candidates for the role of endogenous SOC, although investigations of its functions have generated considerable controversy. In order to assess the role of CaT1 in generating endogenous store-operated Ca(2+) current (I(SOC)) in the lymph node carcinoma of the prostate (LNCaP) human prostate cancer epithelial cell line, we manipulated its endogenous levels by means of antisense hybrid depletion or pharmacological up-regulation (antiandrogen treatment) combined with functional evaluation of I(SOC). Antisense hybrid depletion of CaT1 decreased I(SOC) in LNCaP cells by approximately 50%, whereas enhancement of CaT1 levels by 60% in response to Casodex treatment potentiated I(SOC) by 30%. The functional characteristics of I(SOC) in LNCaP cells were similar in many respects to those reported for heterologously expressed CaT1, although 2-aminoethoxydiphenyl borate sensitivity and lack of constitutive current highlighted notable departures. Our results suggest that CaT1 is definitely involved in I(SOC), but it may constitute only a part of the endogenous SOC, which in general may be a heteromultimeric channel composed of homologous CaT1 and other transient receptor potential subunits. PMID- 12584204 TI - Sphingosine-1-phosphate lyase is involved in the differentiation of F9 embryonal carcinoma cells to primitive endoderm. AB - Sphingosine 1-phosphate (S1P) is a bioactive lipid molecule that acts both extracellularly and intracellularly. The SPL gene encodes a mammalian S1P lyase that degrades S1P. Here, we have disrupted the SPL gene in mouse F9 embryonal carcinoma cells by gene targeting. This is the first report of gene disruption of mammalian S1P lyase. The SPL-null cells exhibited no S1P lyase activity, and intracellular S1P was increased approximately 2-fold, compared with wild-type cells. Treatment of F9 embryonal carcinoma cells with retinoic acid induces differentiation to primitive endoderm (PrE). An acceleration in this PrE differentiation was observed in the SPL-null cells. This effect was apparently caused by the accumulated S1P, since N,N-dimethylsphingosine, a S1P synthesis inhibitor, had an inhibitory effect on the PrE differentiation. Moreover, F9 cells stably expressing sphingosine kinase also exhibited an acceleration in the differentiation. Exogenous S1P had no effect on differentiation, indicating that intracellular but not extracellular S1P is involved. Moreover, we determined that expression of the SPL protein is up-regulated during the progression to PrE. We also showed that sphingosine kinase activity is increased in PrE-differentiated cells. These results suggest that intracellular S1P has a role in the PrE differentiation and that SPL may be involved in the regulation of intracellular S1P levels during this differentiation. PMID- 12584205 TI - 15d-PGJ2 and rosiglitazone suppress Janus kinase-STAT inflammatory signaling through induction of suppressor of cytokine signaling 1 (SOCS1) and SOCS3 in glia. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma agonists are now emerging as therapeutic drugs for various inflammatory diseases. However, their molecular mechanism of action remains to be elucidated. Here we report a novel mechanism that underlies the PPAR-gamma agonist-mediated suppression of brain inflammation. We show that 15-deoxy-Delta12,14-prostaglandin J(2) (15d-PGJ(2)) and rosiglitazone reduce the phosphorylation of STAT1 and STAT3 as well as Janus kinase 1 (JAK1) and JAK2 in activated astrocytes and microglia. The PPAR-gamma agonist-mediated reduction in phosphorylation leads to the suppression of JAK STAT-dependent inflammatory responses. The effects of 15d-PGJ(2) and rosiglitazone are not mediated by activation of PPAR-gamma. 15d-PGJ(2) and rosiglitazone rapidly induce the transcription of suppressor of cytokine signaling (SOCS) 1 and 3, which in turn inhibit JAK activity in activated glial cells. In addition, Src homology 2 domain-containing protein phosphatase 2 (SHP2), another negative regulator of JAK activity, is also involved in their anti-inflammatory action. Our data suggest that 15d-PGJ(2) and rosiglitazone suppress the initiation of JAK-STAT inflammatory signaling independently of PPAR gamma, thus attenuating brain inflammation. PMID- 12584206 TI - Are proximal colorectal cancers always associated with distal adenomas? PMID- 12584207 TI - Lessons from liver transplantation: flip, flop, and why? PMID- 12584208 TI - Gastric inhibitory polypeptide links overnutrition to obesity. PMID- 12584209 TI - Protagonist: population based endoscopic screening for colorectal cancer. PMID- 12584210 TI - Antagonist: population based endoscopic screening for colorectal cancer. PMID- 12584212 TI - Argon plasma coagulation for successful treatment of early gastric cancer with intramucosal invasion. AB - BACKGROUND: In recent years, there has been an increasing number of cases of early gastric cancer (T1, NX) with intramucosal invasion, which are untreatable by surgical or endoscopic mucosal resection (EMR) because of their high risk. Currently, no adequate treatment is available for such patients. AIM: The main objective of this study was to evaluate whether argon plasma coagulation (APC) is an effective and safe modality for treating early gastric cancer untreatable by surgical resection or EMR. METHODS: The study group comprised 20 men and seven women diagnosed with gastric cancer with intramucosal invasion who were considered poor candidates for surgical resection or EMR due to risk factors such as severe cardiac failure or thrombocytopenia. Irradiation conditions for APC treatment were determined using swine gastric mucosa. We used an argon gas flow of 2 l/min at a power setting of 60 W and a maximum irradiation time of 15 s/cm(2). The follow up period of the 27 patients ranged from 18 to 49 months (median 30 months). RESULTS: All lesions were irradiated easily, including areas anatomically difficult for EMR such as the gastric cardia or the posterior wall of the upper gastric body. In 26 of 27 patients (96%) there was no evidence of recurrence during the follow up period (median 30 months). One patient showed recurrence six months after the treatment but was successfully retreated. No serious complications were found in any of the 27 patients but three patients (11%) experienced a feeling of abdominal fullness. INTERPRETATION: APC is a safe and effective modality for treatment of early gastric cancer with intramucosal invasion untreatable by surgical resection or EMR. However, further observations are necessary to determine the long term prognosis of patients undergoing this treatment. PMID- 12584211 TI - Characterisation of telomerase immortalised normal human oesophageal squamous cells. AB - BACKGROUND AND AIMS: Oesophageal cell lines derived from malignancies have numerous genetic abnormalities and therefore are of limited value for studying the early events in carcinogenesis. Reported attempts to establish normal human oesophageal cell lines either have failed to achieve immortalisation or have achieved it by disrupting important cell functions. We have used telomerase technology to establish normal human oesophageal cell lines. METHODS: Endoscopic biopsy specimens of normal oesophageal squamous epithelium were trypsinised, dispersed into single cell suspensions, and cocultivated with ATCC Swiss 3T3 cells. Oesophageal cells were infected with the catalytic subunit of human telomerase (hTERT) using a defective retroviral vector. The integrity of cell cycle checkpoints was tested by measuring p53 response to UV irradiation, and p16 response to infection with H-RasGV12. Expression of a differentiation marker was tested by measuring involucrin response to calcium exposure. RESULTS: Cultures of uninfected oesophageal cells had weak telomerase activity at baseline but exhibited loss of telomerase activity and progressive telomere shortening before undergoing senescence between population doublings (PD) 40-45. In contrast, hTERT infected cells exhibited sustained telomerase activity and stabilisation of telomere length. These cells have reached PD 100 with no diminution in growth rate, while cell cycle checkpoint integrity and involucrin response to calcium exposure have remained intact. CONCLUSIONS: By introducing telomerase into normal human oesophageal squamous cells cocultivated with feeder layers, we have established a cell line that retains normal cell cycle checkpoints and normal differentiation markers. This cell line may be useful for studying the early events in oesophageal carcinogenesis. PMID- 12584213 TI - A rapid decrease in the expression of DMT1 and Dcytb but not Ireg1 or hephaestin explains the mucosal block phenomenon of iron absorption. AB - BACKGROUND: A large oral dose of iron will reduce the absorption of a subsequent smaller dose of iron in a phenomenon known as mucosal block. Molecular analysis of this process may provide insights into the regulation of intestinal iron absorption. AIMS: To determine the effect of an oral bolus of iron on duodenal expression of molecules associated with intestinal iron transport in rats and to relate this to changes in iron absorption. METHODS: Rats were given an oral dose of iron and duodenal expression of divalent metal transporter 1 (DMT1), Dcytb, Ireg1, and hephaestin (Hp) was determined using the ribonuclease protection assay, western blotting, and immunofluorescence. Iron absorption was measured using radioactive (59)Fe. RESULTS: A decrease in intestinal iron absorption occurred following an oral dose of iron and this was associated with increased enterocyte iron levels, as assessed by iron regulatory protein activity and immunoblotting for ferritin. Reduced absorption was also accompanied by a rapid decrease in expression of the mRNAs encoding the brush border iron transport molecules Dcytb and the iron responsive element (IRE) containing the splice variant of DMT1. No such change was seen in expression of the non-IRE splice variant of DMT1 or the basolateral iron transport molecules Ireg1 and Hp. Similar changes were observed at the protein level. CONCLUSIONS: These data indicate that brush border, but not basolateral, iron transport components are regulated locally by enterocyte iron levels and support the hypothesis that systemic stimuli exert their primary effect on basolateral transport molecules. PMID- 12584214 TI - 18F-fluoro-deoxy-glucose positron emission tomography (18F-FDG-PET) for assessment of enteropathy-type T cell lymphoma. AB - BACKGROUND AND AIMS: Enteropathy-type T cell lymphoma (ETCL) represents a relatively rare disease, accounting for less than 1% of non-Hodgkin's lymphomas. ETCL is an aggressive lymphoma which may either present de novo or arise in the context of longstanding or untreated coeliac disease (CD). The aim of this study was to evaluate the potential of 18F-fluoro-deoxy-glucose positron emission tomography (18F-FDG-PET) for imaging of ETCL. Furthermore, we wished to evaluate whether the presence of CD might provide a potential diagnostic obstacle to imaging of lymphoma due to unspecific 18F-FDG uptake and whether accumulation of 18F-FDG within the gut correlates with activity of CD. PATIENTS AND METHODS: We retrospectively analysed patients with ETCL and individuals suffering from CD undergoing 18F-FDG-imaging at our PET unit. Material for histological reassessment by a reference pathologist had to be available for inclusion of patients in the analysis. Whole body 18F-FDG-PET scans were performed 40 minutes following injection of 300-380 MBq of 18F-FDG. Images were reconstructed iteratively. In areas with focally elevated FDG uptake and in case of diffusely elevated intestinal 18F-FDG accumulation, standard uptake values (SUVs) were calculated. RESULTS: During a period of two years, five patients (one male, four female) with a mean age of 56.4 years (range 44-62) with a diagnosis of ETCL underwent 18F-FDG-PET. Four of these patients were imaged before application of cytotoxic treatment while one patient had regular PET scans for follow up. All four patients undergoing pre-therapeutic imaging showed markedly elevated intestinal 18F-FDG uptake, with a maximal SUV of 6.4-8.0 (mean 7.15 (SD 0.82)). The patient imaged following surgery and cytotoxic therapy had no pathologic 18F FDG uptake which was found to correlate with normal duodenal mucosa, as evidenced by repeated biopsies and conventional imaging methods. During the same time span, 12 patients (five male, seven female) with a mean age of 63.8 years (range 42-82) suffering from CD were imaged. Four of these patients showed no elevated intestinal 18F-FDG uptake while five had minor diffuse intestinal 18F-FDG accumulation with SUVs ranging between 2.2 and 4.6 (mean 3.4 (SD 0.89)). In the remaining three patients with diffuse intestinal 18F-FDG uptake, no SUV could be calculated. SUVs in patients with ETCL were remarkably higher than in patients suffering from CD (p=0.011), irrespective of the activity of CD at the time of imaging. CONCLUSION: In spite of the relatively small number of patients, our results clearly indicate the potential value of 18F-FDG-PET for diagnosing and imaging ETCL. In addition, the data also suggest that 18F-FDG-PET may lead to early diagnosis in individuals developing ETCL in the context of longstanding CD. This is due to the fact that 18F-FDG does not appear to significantly accumulate in the gut of patients with CD, irrespective of disease activity. PMID- 12584215 TI - Scintigraphy versus manometry in patients with suspected biliary sphincter of Oddi dysfunction. AB - INTRODUCTION: Sphincter of Oddi (SO) manometry is at present the "gold standard" investigation for patients with suspected biliary SO dysfunction. Non-invasive scintigraphy in cholecystectomised patients using a complex scoring system or the transit time from the hepatic hilum to the duodenum (HDTT) have been promoted as sensitive and specific alternatives. AIM: To evaluate the scintigraphic scoring system and HDTT in patients with suspected biliary SO dysfunction undergoing SO manometry. METHODS: Cholecystectomised patients undergoing SO manometry for persistent biliary-type pain, as defined by the Rome II criteria, for which all other causes had been excluded, were prospectively studied. Scintigraphy with cholecystokinin octapeptide infusion was performed within a month prior to manometry. Scoring of the scans and measurement of HDTT was performed by independent blinded observers. Manometry of the biliary sphincter was performed per-endoscopically and defined as abnormal if basal pressure was > or = 40 mm Hg. RESULTS: Thirty two patients were enrolled (30 females, mean age 45.1 years). Three patients were excluded from analysis because manometry from the bile duct was not technically possible. Eight patients had abnormal manometry. Scintigraphic scoring had a sensitivity of 25-38%, a specificity of 86-89%, positive predictive value (PPV) of 40-60%, and a negative predictive value (NPV) of 75-79%. The coefficient of variation for interobserver variation in scores was 0.72. HDTT sensitivity was 13%, specificity 95%, PPV 50%, and NPV 74%. CONCLUSIONS: Our findings indicate that scintigraphy using these methods of analysis correlates poorly with manometry in post cholecystectomy patients with suspected biliary SO dysfunction. PMID- 12584216 TI - Insurance problems among inflammatory bowel disease patients: results of a Dutch population based study. AB - BACKGROUND AND AIMS: The majority of patients with inflammatory bowel disease (IBD) have a normal life expectancy and therefore should not be weighted when applying for life assurance. There is scant literature on this topic. In this study our aim was to document and compare the incidence of difficulties in application for life and medical insurance in a population based cohort of IBD patients and matched population controls. METHODS: A population based case control study of 1126 IBD patients and 1723 controls. Based on a detailed questionnaire, the frequency and type of difficulties encountered when applying for life and medical insurance in matched IBD and control populations were appraised. RESULTS: In comparison with controls, IBD patients had an 87-fold increased risk of encountering difficulties when applying for life assurance (odds ratio (OR) 87 (95% confidence interval (CI) 31-246)), with a heavily weighted premium being the most common problem. Patients of high educational status, with continuous disease activity, and who smoked had the highest odds of encountering such problems. Medical insurance difficulties were fivefold more common in IBD patients compared with controls (OR 5.4 (95% CI 2.3-13)) although no specific disease or patient characteristics were identified as associated with such difficulties. CONCLUSIONS: This is the first detailed case control study that has investigated insurance difficulties among IBD patients. Acquiring life and medical insurance constituted a major problem for IBD patients in this study. These results are likely to be more widely representative given that most insurance companies use international guidelines for risk assessment. In view of the recent advances in therapy and promising survival data on IBD patients, evidence based guidelines for risk assessment of IBD patients by insurance companies should be drawn up to prevent possible discriminatory practices. PMID- 12584217 TI - Local delivery of adenoviral vectors encoding murine interleukin 10 induces colonic interleukin 10 production and is therapeutic for murine colitis. AB - INTRODUCTION: Interleukin 10 knockout (IL-10-/-) mice spontaneously develop a Th1 T cell mediated colitis with many similarities to Crohn's disease. Daily injections of IL-10 are unable to induce remission in mice with established disease. In contrast, we have shown previously that intravenous administration of adenoviral vectors encoding IL-10 (AdvmuIL-10) induces hepatic IL-10 release and leads to long term disease suppression with profound systemic immunoregulatory changes. AIMS: To determine whether rectal delivery of AdvmuIL-10 induces localised colonic IL-10 expression without systemic immune suppression, and assess its therapeutic efficacy in IL-10-/- mice with established colitis. RESULTS: A single rectal infusion of 5 x 10(8) PFU AdvmuIL-10 to 10 week IL-10-/- mice resulted in a median level of 27.3 pg/mg IL-10 in colonic homogenates harvested one week later. IL-10-/- mice with established colitis treated with an enema of 5 x 10(8) PFU AdvmuIL-10 entered clinical and histological remission whereas empty cassette adenovirus (Adv0) or phosphate buffered saline (PBS) treated mice developed progressive disease. After four weeks, the histological score of AdvmuIL-10 treated mice (4.4 (1.5)) was significantly lower than that of Adv0 (11.1 (1.1); p<0.001) and PBS (10.9 (1.0); p<0.01) treated controls. In addition, the stool concentration of IL-1 beta over the four week experiment was significantly higher in mice treated with saline or Adv0 than in those treated with AdvmuIL-10 (p<0.01). CONCLUSION: Local AdvmuIL-10 therapy reverses colitis in IL-10-/- mice without the systemic effects seen after intravenous administration. Gene therapy strategies using adenoviral vectors encoding immunoregulatory cytokines may prove to be a potent approach to the treatment of chronic inflammatory diseases such as Crohn's disease. PMID- 12584218 TI - Lactobacillus GG prevents recurrence of colitis in HLA-B27 transgenic rats after antibiotic treatment. AB - BACKGROUND AND AIMS: Bacteroides vulgatus induces colitis in gnotobiotic HLA-B27 transgenic (TG) rats while broad spectrum antibiotics prevent and treat colitis in specific pathogen free (SPF) TG rats although disease recurs after treatment ends. Lactobacilli treat human pouchitis and experimental colitis. We investigated if Lactobacillus rhamnosus GG (L GG) can prevent colitis in TG rats monoassociated with B vulgatus and if L GG or Lactobacillus plantarum 299v (LP 299v) can treat established colitis in SPF TG rats and prevent recurrent disease after antibiotics were stopped. METHODS: Germfree B27 TG rats were monoassociated with B vulgatus for four weeks following two weeks of colonisation with L GG or no bacteria. SPF B27 TG rats received oral vancomycin and imipenem for two weeks, or water alone, followed by four weeks of treatment with oral L GG, LP 299v, or water only. Disease activity was quantified by blinded gross and histological scores, caecal myeloperoxidase (MPO) activity, and levels of interleukin (IL)-1 beta, tumour necrosis factor (TNF), transforming growth factor beta, and IL-10. RESULTS: L GG did not prevent colitis in B vulgatus co-associated TG rats or treat established disease in SPF rats. However, L GG but not LP 299v prevented colitis relapse in antibiotic treated rats with reduced gross and histological scores, caecal MPO, IL-1 beta, and TNF whereas caecal IL-10 was increased. CONCLUSIONS: L GG does not prevent colitis in gnotobiotic TG rats or treat established disease in SPF rats, but is superior to LP 299v in the prevention of recurrent colitis. These studies suggest that antibiotics and probiotic agents provide synergistic therapeutic effects, perhaps mediated by altered immunomodulation with selective activity of different lactobacillus species. PMID- 12584219 TI - Safety and efficacy of intravenous pulse cyclophosphamide in acute steroid refractory inflammatory bowel disease. AB - BACKGROUND AND AIMS: One major problem in the management of steroid refractory attacks of patients with inflammatory bowel disease (IBD) is the establishment of a rapidly acting immunosuppressive regimen. Based on its well known efficacy in systemic vasculitis, intravenous cyclophosphamide pulse therapy was used in refractory IBD patients to evaluate both its efficacy and safety. METHODS: Between December 1998 and May 2001, seven patients (Crohn's disease, n=5; indeterminate colitis, n=1) with severe steroid refractory IBD (Crohn's disease activity index (CDAI) 264-479 points) received 4-6 cycles of monthly treatment with intravenous cyclophosphamide (750 mg) in a prospective uncontrolled pilot study. RESULTS: All patients improved after two intravenous pulses of cyclophosphamide and six of seven patients achieved complete remission (CDAI <150 points). One patient with Crohn's disease of the small and large bowel showed an impressive clinical response but did not enter into remission. Tapering to low dose steroids was possible in all responders. Remission was maintained in all patients for 18 months (median) but required a second course of intravenous pulse cyclophosphamide in one patient. The drug was well tolerated except for two episodes of candida oesophagitis. CONCLUSIONS: Intravenous pulse cyclophosphamide may be a safe and effective treatment in patients with severe IBD unresponsive to steroid treatment and merits evaluation in a controlled trial. PMID- 12584220 TI - Isolation, maturational level, and functional capacity of human colon lamina propria plasma cells. AB - BACKGROUND AND AIMS: Large numbers of plasma cells (PC) localise in the intestinal lamina propria (LP) where they play a critical role in the defence against pathogens. This study analyses the level of maturation reached by normal human colon LPPC in comparison with that of bone marrow (BM) PC. METHODS: A technique was designed to purify LPPC by combining collagenase digestion of the mucosal layer and immunomagnetic selection of CD54(+) LP cells. It provided highly purified PC, as demonstrated by morphology, CD38(h) phenotype, and cytoplasmic IgA staining criteria. This procedure allowed comparison of in vitro functional capacities and a broad phenotypic analysis of BMPC and LPPC. RESULTS: LPPC and BMPC exhibited identical expression of differentiation markers (CD19( /+), CD20(-), HLA-DR(low/-), VS38c(high)), survival molecules (CD95 (low/-), Bcl 2(+)), and B cell transcription factor profile, as well as similar in vitro Ig secreting kinetics (14 days) and lack of susceptibility to apoptosis by CD95 ligation. In contrast, they markedly differed in adhesion molecule expression, as LPPC showed higher levels of CD44 and CD21 and were alpha 4 beta 7(+) whereas BMPC lacked this integrin and expressed higher levels of CD49d and CD31. CONCLUSION: These data indicate that PC at effector sites of the humoral response (BM and LP) show similar high differentiation, survival, and functional features but display a distinctive pattern of adhesion molecules, probably related to their respective homing locations. PMID- 12584221 TI - Diagnosing small bowel Crohn's disease with wireless capsule endoscopy. AB - BACKGROUND: The small bowel is the most commonly affected site of Crohn's disease (CD) although it may involve any part of the gastrointestinal tract. The current methodologies for examining the small bowel are x ray and endoscopy. AIMS: To evaluate, for the first time, the effectiveness of wireless capsule endoscopy in patients with suspected CD of the small bowel undetected by conventional modalities, and to determine the diagnostic yield of the M2A Given Capsule. PATIENTS: Seventeen patients (eight males, mean age 40 (15) years) with suspected CD fulfilled study entry criteria: nine had iron deficiency anaemia (mean haemoglobin 10.5 (SD 1.8) g%), eight had abdominal pain, seven had diarrhoea, and three had weight loss. Small bowel x ray and upper and lower gastrointestinal endoscopic findings were normal. Mean duration of symptoms before diagnosis was 6.3 (SD 2.2) years. METHODS: Each subject swallowed an M2A Given Capsule containing a miniature video camera, batteries, a transmitter, and an antenna. Recording time was approximately eight hours. The capsule was excreted naturally in the patient's bowel movement, and the data it contained were retrieved and interpreted the next day. RESULTS: Of the 17 study participants, 12 (70.6%, six males, mean age 34.5 (12) years) were diagnosed as having CD of the small bowel according to the findings of the M2A Given Capsule. CONCLUSIONS: Wireless capsule endoscopy diagnosed CD of the small bowel (diagnostic yield of 71%). It was demonstrated as being an effective modality for diagnosing patients with suspected CD undetected by conventional diagnostic methodologies. PMID- 12584222 TI - Contrast enhanced magnetic resonance imaging of the terminal ileum in children with Crohn's disease. AB - BACKGROUND AND AIMS: Recently, magnetic resonance imaging (MRI) has been introduced in the diagnosis of patients with inflammatory bowel disease (IBD). However, it is still rarely reported in paediatric IBD. We studied the diagnostic value of gadolinium enhanced MRI in revealing inflammation of the distal ileum in children with Crohn's disease (CD) and in differentiating them from patients with other inflammatory diseases of the gut. MRI was performed using a polyethylene glycol (PEG) solution as oral contrast agent to distend the small bowel (CE-PEG MRI). SUBJECTS AND METHODS: Seventy five consecutive patients (median age 13.6 years, range 8-17) with suspected CD underwent ileocolonoscopy with biopsy and CE PEG-MRI. CD activity was measured by the paediatric Crohn's disease activity index (PCDAI). CE-PEG-MRI was evaluated with an overall score calculated, taking into account both wall thickness and contrast enhancement. RESULTS: Active CD with distal ileitis was diagnosed in 26 cases, active ulcerative colitis (UC) in 18, and spondyloarthropathy and indeterminate ileocolitis in 11; 20 children served as controls. In all CD patients, CE-PEG-MRI revealed a marked ileal involvement with increased wall thickness and parietal contrast enhancement and showed a high concordance with endoscopy and histology, whereas the test was negative in all controls. Of the 18 UC patients, CE-PEG-MRI was negative in 15 and showed a mild parietal contrast enhancement of the terminal ileum in only three of seven patients with backwash ileitis. Among the group of spondyloarthropathy patients, six had mucosal erosions and five mild superficial ileitis: CE-PEG-MRI was negative in four and revealed only mild parietal contrast enhancement of the ileal wall in seven. CE-PEG-MRI did not show an increase in wall thickness of the distal ileum in any of the UC or spondyloarthropathy patients. The sensitivity and specificity of CE-PEG-MRI related to the presence of erosive ileitis, as documented by endoscopy, were 84% and 100%, respectively. In addition, the test correlated markedly with endoscopy and histology in the entire population (r=0.94; r=0.95, respectively) as well as with the PCDAI in CD patients (r=0.91). CONCLUSIONS: In children with active CD, CE-PEG-MRI is a very sensitive and specific test for the detection of distal ileitis and for differentiation from other inflammatory diseases of the gut. The test could also be useful for the firstline diagnostic approach in children with suspected CD. The high correlation of CE-PEG-MRI with ileal endoscopy and histology as well as with PCDAI makes this test of great interest for future studies as a tool for monitoring the clinical course and the effect of therapy in CD patients. PMID- 12584223 TI - Grading of distal colorectal adenomas as predictors for proximal colonic neoplasia and choice of endoscope in population screening: experience from the Norwegian Colorectal Cancer Prevention study (NORCCAP). AB - BACKGROUND AND AIMS: The purpose of this study was to evaluate the utility of easily measured clinical variables at flexible sigmoidoscopy (FS) screening that might predict a proximal advanced neoplasm (PAN). METHODS: We studied 1833 subjects with biopsy verified adenomas at FS who subsequently underwent full colonoscopy. RESULTS: A total of 387 (21%) subjects had proximal colonic neoplasms (PCN) and 85 (5%) had PAN. In univariate comparison, the risk of PAN increased more than threefold in the presence of a distal adenoma measuring either > or =10 mm in diameter or containing villous components. Multiplicity of distal adenomas, severe dysplasia, or age > or =60 years increased the risk of PAN more than twofold. In the multivariate model, the presence of a distal adenoma > or =10 mm, villousness, and multiplicity maintained their significance as predictive variables for increased risk of proximal neoplasms, whereas sex and severe dysplasia lost their significance. By recommending colonoscopy only to individuals with multiple (>1) adenomas or any high risk adenoma at FS, we would have reduced the number of colonoscopies by 1209 (66%) but would have missed 32 (38%) participants with PAN and 217 (56%) with PCN. By using a 60 cm endoscope instead of an ordinary colonoscope at FS, nine (2%) participants with advanced neoplasms, including three patients with cancer, would have been missed. CONCLUSION: The present study supports the concept of defining "any adenoma" as a positive FS, qualifying for colonoscopy. We recommend the use of an ordinary colonoscope instead of a 60 cm sigmoidoscope for FS screening examinations. PMID- 12584224 TI - Thrombotic risk factors and extent of liver fibrosis in chronic viral hepatitis. AB - BACKGROUND AND AIMS: Thrombosis of the small intrahepatic veins has been suggested to trigger liver tissue remodelling. We evaluated the prevalence of multiple thrombotic risk factors and their association with the extent of fibrosis in chronic viral hepatitis. METHODS: Ninety consecutive patients with chronic hepatitis B or C without malignancy, a history of venous thrombosis, or antiviral/immunosuppressive therapy within the last six months were included. Thrombophilic and coagulation factors were evaluated on the liver biopsy day. RESULTS: One or more thrombotic risk factors were found in 68% and > or =2 factors in 37% of patients. Higher necroinflammatory activity was independently associated with higher prothrombin time (p=0.003), alanine aminotransferase level (p=0.011), and histological staging (p=0.018). Patients with staging scores of 4 6 compared with those with scores of 0-3 more frequently had deficiency of protein C (24% v 3%; p=0.007), antithrombin III (28% v 5%; p=0.005), and plasminogen (19% v 2%; p=0.03), and a trend for more frequent activated protein C resistance (8% v 0%; p=0.075). The presence of > or =1 significant thrombotic risk factor was observed in 11/25 (44%) patients with staging scores of 4-6 and in 6/65 (9%) patients with scores of 0-3 (p<0.001), being the only variable independently associated with advanced staging (odds ratio 2.4, p=0.02). CONCLUSIONS: Thrombotic risk factors are frequently detected in patients with chronic viral hepatitis and the presence of > or =1 significant factor is associated with more advanced fibrosis. Whether the association of such thrombophilic conditions with advanced fibrosis is a primary or secondary phenomenon and whether their development in combination with local inflammation accelerate the progression of liver fibrosis need further evaluation. PMID- 12584225 TI - Enhanced vasodilatation to endothelin antagonism in patients with compensated cirrhosis and the role of nitric oxide. AB - BACKGROUND AND AIMS: Patients with advanced cirrhosis have systemic vasodilatation and increased nitric oxide (NO) production despite activated vasopressor systems, including the endothelin system. The aims of this study were to assess the contribution of endogenous endothelin 1 (ET-1) and NO to the maintenance of basal forearm vascular tone in patients with preascitic cirrhosis (n=7) and in age and sex matched healthy controls (n=7). METHODS: Using venous occlusion plethysmography, forearm blood flow (FBF) responses to subsystemic locally active intra-arterial infusion of BQ-123 (a selective endothelin type A receptor (ET(A)) receptor antagonist; 10 nmol/min) were measured before and during application of an "NO clamp": a balanced co-infusion of L-N(G)-monomethyl arginine (a selective NO synthase inhibitor) and sodium nitroprusside (an exogenous NO donor) to block endogenous NO production and restore NO mediated basal FBF, respectively. RESULTS: L-NMMA infusion produced a reduction in FBF (p<0.001) which was similar in both groups. Before applying the "NO clamp", BQ 123 caused an increase in FBF in both groups (p<0.001) that was greater in patients with cirrhosis (p<0.01). During the "NO clamp", BQ-123 induced vasodilatation was abolished in controls and attenuated in patients (p<0.001) but remained significantly greater in patients with cirrhosis (p<0.01). CONCLUSIONS: These findings indicate a greater ET(A) mediated contribution of endogenous ET-1 to the maintenance of basal forearm vascular tone in patients with preascitic cirrhosis. In addition, enhanced vasodilatation to ET(A) receptor antagonism in cirrhosis cannot be entirely attributed to NO release but is likely to be related to reversal of direct ET-1 mediated tone. PMID- 12584226 TI - A large population study of spontaneous HBeAg seroconversion and acute exacerbation of chronic hepatitis B infection: implications for antiviral therapy. AB - BACKGROUND AND AIM: Clinical data on spontaneous hepatitis B e antigen (HBeAg) seroconversion and acute exacerbation of chronic hepatitis B (CHB) virus infection from large population studies are lacking. In the present study we examined the clinical features and significance of HBeAg seroconversion and acute exacerbation in 3063 Chinese CHB patients. METHODS: Clinical assessment, liver biochemistry, hepatitis B virus (HBV) serology and HBV DNA, time of HBeAg seroconversion, and acute exacerbation were monitored. RESULTS: Median age at HBeAg seroconversion was 34.5 years. The cumulative HBeAg seroconversion rate significantly increased with alanine aminotransferase (ALT) levels on presentation (p<0.0001). For patients with ALT levels more than twice the upper limit of normal (ULN) on presentation, the HBeAg seroconversion rate at the fifth year of follow up was 72.4%. After HBeAg seroconversion, 65.2% (73/110) of patients had undetectable HBV DNA levels by the Digene Hybrid Capture assay. Of these, 78.1% still had HBV DNA levels detectable by the Amplicor HBV Monitor Test. We found that 37.5% antibody to HBeAg (anti-HBe) positive patients had undetectable HBV DNA levels by the Digene Hybrid Capture assay before acute exacerbation. Acute exacerbations of longer duration, with higher peak ALT, bilirubin, and alpha fetoprotein levels were associated with an increased HBeAg seroconversion rate (p<0.0001-0.045). Acute exacerbation with peak ALT levels more than five times the ULN carried a 46.4% chance of HBeAg seroconversion within three months. HBeAg seroreversion and mortality occurred in 2.7% and 0.7% of acute exacerbations, respectively. CONCLUSION: In the present study we have provided information on HBeAg seroconversion and acute exacerbation, which are important in decision making for CHB treatment and in designing clinical trials. PMID- 12584227 TI - Durability of HBeAg seroconversion following antiviral therapy for chronic hepatitis B: relation to type of therapy and pretreatment serum hepatitis B virus DNA and alanine aminotransferase. AB - BACKGROUND AND AIMS: Interferon (IFN) induced hepatitis B e antigen (HBeAg) seroconversion is durable in 80-90% of chronic hepatitis B patients. Preliminary reports on the durability of HBeAg seroconversion following lamivudine are contradictory. We investigated the durability of response following IFN, lamivudine, or IFN-lamivudine combination therapy in a meta-analysis of individual patient data. PATIENTS AND METHODS: Twenty four centres included 130 patients in total with an HBeAg seroconversion (HBeAg negative, antibodies to hepatitis B e antigen positive) at the end of antiviral therapy: 59 with lamivudine, 49 with interferon, and 22 with combination therapy. Relapse was defined as confirmed reappearance of HBeAg. RESULTS: The three year cumulative HBeAg relapse rate by the Kaplan-Meier method was 54% for lamivudine, 32% for IFN, and 23% for combination therapy (p=0.01). Cox regression analysis identified pretreatment hepatitis B virus (HBV) DNA, alanine aminotransferase (ALT), sex, and therapy as independent predictive factors of post-treatment relapse; Asian race, previous therapy, centre, and type of study were not predictive of relapse. The relative HBeAg relapse risk of lamivudine compared with IFN therapy was 4.6 and that of combination therapy to IFN therapy 0.7 (p(overall)=0.01). CONCLUSIONS: The durability of HBeAg seroconversion following lamivudine treatment was significantly lower than that following IFN or IFN-lamivudine combination therapy. The risk of relapse after HBeAg seroconversion was also related to pretreatment levels of serum ALT and HBV DNA, but independent of Asian race. PMID- 12584228 TI - Cost effectiveness of peginterferon alpha-2b plus ribavirin versus interferon alpha-2b plus ribavirin for initial treatment of chronic hepatitis C. AB - BACKGROUND: Peginterferon alpha-2b plus ribavirin therapy in previously untreated patients with chronic hepatitis C yields the highest sustained virological response rates of any treatment strategy but is expensive. AIMS: To estimate the cost effectiveness of treatment with peginterferon alpha-2b plus ribavirin compared with interferon alpha-2b plus ribavirin for initial treatment of patients with chronic hepatitis C. METHODS: Individual patient level data from a randomised clinical trial with peginterferon plus ribavirin were applied to a previously published and validated Markov model to project lifelong clinical outcomes. Quality of life and economic estimates were based on German patient data. We used a societal perspective and applied a 3% annual discount rate. RESULTS: Compared with no antiviral therapy, peginterferon plus fixed or weight based dosing of ribavirin increased life expectancy by 4.2 and 4.7 years, respectively. Compared with standard interferon alpha-2b plus ribavirin, peginterferon plus fixed or weight based dosing of ribavirin increased life expectancy by 0.5 and by 1.0 years with incremental cost effectiveness ratios of 11,800 euros and 6600 euros per quality adjusted life year (QALY), respectively. Subgroup analyses by genotype, viral load, sex, and histology showed that peginterferon plus weight based ribavirin remained cost effective compared with other well accepted medical treatments. CONCLUSIONS: Peginterferon alpha-2b plus ribavirin should reduce the incidence of liver complications, prolong life, improve quality of life, and be cost effective for the initial treatment of chronic hepatitis C. PMID- 12584229 TI - Heterozygous recipient and donor HFE mutations associated with a hereditary haemochromatosis phenotype after liver transplantation. AB - We observed the development of phenotypic hereditary haemochromatosis in a non hereditary haemochromatosis liver transplant recipient, following transplantation with a liver from a C282Y heterozygous donor. No cause for secondary iron overload was identified. Subsequent sequencing of the HFE gene of both donor and recipient revealed a strong candidate for a novel pathogenic HFE mutation. In the recipient, heterozygosity for a single base substitution in exon 1, g.18 G>C, resulting in the substitution of arginine by serine at codon 6 (R6S), was detected. This R6S variation is likely to represent a novel pathogenic missense mutation of the HFE gene. An interaction between R6S heterozygosity in the recipient and C282Y heterozygosity in the donor liver is the most likely explanation for the development of iron overload in this patient. The report suggests that an hepatic defect is required for expression of hereditary haemochromatosis and that the intestinal HFE genotype is not the exclusive determinant of iron status. It also raises the possibility that a hereditary haemochromatosis phenotype may result from transplantation of C282Y heterozygous donor livers into recipients with heterozygous pathogenic HFE mutations. This possibility may have significant implications for the common practice of transplanting C282Y heterozygous livers. PMID- 12584230 TI - Probiotics for infectious diarrhoea. PMID- 12584231 TI - Budd-Chiari syndrome with portal, splenic, and superior mesenteric vein thrombosis treated with TIPS: who dares wins. PMID- 12584233 TI - Caerulein induced pancreatitis. PMID- 12584234 TI - Osteoporosis and liver disease: additional reasons for coeliac disease screening. PMID- 12584235 TI - Pathology and cost effectiveness of endoscopy surveillance for premalignant gastric lesions. PMID- 12584236 TI - Adult height in patients with early onset of Crohn's disease. PMID- 12584232 TI - Intestinal epithelial responses to enteric pathogens: effects on the tight junction barrier, ion transport, and inflammation. AB - The effects of pathogenic organisms on host intestinal epithelial cells are vast. Innumerable signalling pathways are triggered leading ultimately to drastic changes in physiological functions. Here, the ways in which enteric bacterial pathogens utilise and impact on the three major physiological functions of the intestinal epithelium are discussed: alterations in the structure and function of the tight junction barrier, induction of fluid and electrolyte secretion, and activation of the inflammatory cascade. This field of investigation, which was virtually non-existent a decade ago, has now exploded, thus rapidly expanding our understanding of bacterial pathogenesis. Through increased delineation of the ways in which microbes alter host physiology, we simultaneous gain insight into the normal regulatory mechanisms of the intestinal epithelium. PMID- 12584237 TI - Gastrointestinal epithelial neoplasia. PMID- 12584238 TI - Underdiagnosis of hereditary haemochromatosis: reflects lack of clinical not biochemical penetrance. PMID- 12584239 TI - Stem cells, niches and cadherins: a view from Drosophila. AB - Stem cells are essential for the correct development and homeostasis of adult organisms, as well as having obvious potential therapeutic importance. Analysis of the biology of stem cells and their regulatory microenvironment in adult organs has, however, been hindered by the rarity of these cells in mature tissues and by the lack of positive markers for them. The ovary of the Drosophila melanogaster female is a stem cell niche in which such analyses can be performed. The stromal cells of the microenvironment act as a regulatory centre to control the proliferation and differentiation of the germline stem cells, using several signalling molecules, among them the protein DPP - a Drosophila homologue of the human bone morphogenetic proteins BMP2 and BMP4. Recent work shows that DE cadherin-mediated adhesion is used for the initial recruitment and posterior anchoring of the germline-derived stem cells in their niche. PMID- 12584240 TI - Megalin and the neurodevelopmental biology of sonic hedgehog and retinol. AB - Megalin is a receptor expressed by embryonic epithelia that mediates endocytosis of numerous ligands, including sonic hedgehog (Shh) and retinol, the precursor to retinoic acid (RA). The importance of Shh and RA signaling in neurodevelopment, combined with the fact that megalin-deficient mice show profound neurodevelopmental abnormalities, has raised questions as to the possible role of megalin in Shh and RA signaling. Several mechanisms could explain how megalin influences Shh and RA signaling in the context of neurodevelopment. These include the involvement of megalin in the transport of Shh and retinol within neuroepithelia, as well as direct signal transduction as a response to binding of Shh and retinol to megalin. PMID- 12584241 TI - Dynamic relocalization of the chromosomal passenger complex proteins inner centromere protein (INCENP) and aurora-B kinase during male mouse meiosis. AB - INCENP and aurora-B kinase are two chromosomal passenger proteins that are thought to play key roles in coordinating chromosome segregation with cytokinesis in somatic cells. Here we have analyzed their subcellular distribution, and that of phosphorylated histone H3, and the timing of their relative appearance in mouse spermatocytes during both meiotic divisions. Our results show that in mitotic spermatogonial cells, INCENP and aurora-B show the same pattern of distribution as they do in cultured somatic cells. INCENP labels the synaptonemal complex central element from zygotene up to late pachytene when it begins to relocalize to heterochromatic chromocentres. Aurora-B first appears at chromocentres in late diplotene before the initial phosphorylation of histone H3. INCENP and aurora-B concentrate at centromeres during diakinesis and appear during metaphase I as T-shaped signals at their inner domains, just below associated sister kinetochores. During late anaphase I both proteins relocalize to the spindle midzone. Both proteins colocalize at a connecting strand traversing the centromere region and joining sister kinetochores, in metaphase II centromeres. This strand disappears at the metaphase II/anaphase II transition and relocalizes to the spindle midzone. We discuss the complex dynamic relocalization of the chromosomal passenger complex during prophase I. Additionally, we suggest that this complex may regulate sister-chromatid centromere cohesion during both meiotic divisions. PMID- 12584242 TI - Live-cell monitoring of tyrosine phosphorylation in focal adhesions following microtubule disruption. AB - Tyrosine phosphorylation of focal adhesion components is involved in the regulation of focal adhesion formation and turnover, yet the underlying molecular mechanisms are still poorly defined. In the present study, we have used quantitative fluorescence microscopy to investigate the dynamic relationships between the incorporation of new components into growing focal adhesions and tyrosine phosphorylation of these sites. For this purpose, a new approach for monitoring phosphotyrosine levels in live cells was developed, based on a 'phosphotyrosine reporter' consisting of yellow fluorescent protein fused to two consecutive phosphotyrosine-binding Src-homology 2 (SH2)-domains derived from pp60(c-Src). This YFP-dSH2 localized to cell-matrix adhesions and its intensity was linearly correlated with that of an anti-phosphotyrosine antibody labeling. The differential increase in vinculin and phosphotyrosine levels was examined in live cells by two-color time-lapse movies of CFP-vinculin and YFP-dSH2. In this study, focal adhesion growth was triggered by microtubule disruption, which was previously shown to stimulate focal adhesion development by inducing cellular contraction. We show here that, 2 minutes after addition of the microtubule disrupting drug nocodazole, the local densities of the focal adhesion-associated proteins vinculin, paxillin and focal adhesion kinase (FAK) are significantly elevated and the focal adhesion area is increased, whereas elevation in tyrosine phosphorylation inside the growing adhesions occurs only a few minutes later. Phosphotyrosine and FAK density reach their maximum levels after 10 minutes of treatment, whereas vinculin and paxillin levels as well as focal adhesion size continue to grow, reaching a plateau at about 30 minutes. Our findings suggest that protein recruitment and growth of focal adhesions are an immediate and direct result of increased contractility induced by microtubule disruption, whereas tyrosine phosphorylation is activated later. PMID- 12584243 TI - Alterations in the Arf6-regulated plasma membrane endosomal recycling pathway in cells overexpressing the tetraspan protein Gas3/PMP22. AB - Growth arrest specific 3 (Gas3)/peripheral myelin protein 22 (PMP22) is a component of the compact peripheral nerve myelin, and mutations affecting gas3/PMP22 gene are responsible for a group of peripheral neuropathies in humans. We have performed in vivo imaging in order to investigate in detail the phenotype induced by Gas3/PMP22 overexpression in cultured cells. Here we show that Gas3/PMP22 triggers the accumulation of vacuoles, before the induction of cell death or of changes in cell spreading. Overexpressed Gas3/PMP22 accumulates into two distinct types of intracellular membrane compartments. Gas3/PMP2 accumulates within late endosomes close to the juxtanuclear region, whereas in the proximity of the cell periphery, it induces the formation of actin/phosphatidylinositol (4,5)-bisphosphate (PIP(2))-positive large vacuoles. Gas3/PMP22-induced vacuoles do not contain transferrin receptor, but instead they trap membrane proteins that normally traffic through the ADP-ribosylation factor 6 (Arf6) endosomal compartment. Arf6 and Arf6-Q67L co-localize with Gas3/PMP22 in these vacuoles, and the dominant negative mutant of Arf6, T27N, blocks the appearance of vacuoles in response to Gas3/PMP22, but not its accumulation in the late endosomes. Finally a point mutant of Gas3/PMP22 responsible for the Charcot-Marie-Tooth 1A disease is unable to trigger the accumulation of PIP(2)-positive vacuoles. Altogether these results suggest that increased Gas3/PMP22 levels can alter membrane traffic of the Arf6 plasma-membrane-endosomal recycling pathway and show that, similarly to other tetraspan proteins, Gas3/PMP22 can accumulate in the late endosomes. PMID- 12584244 TI - Cadherin function probed by laser tweezer and single molecule fluorescence in vascular endothelial cells. AB - In endothelial monolayers agonist-induced influx of Ca(2+) and activities of the actin cytoskeleton have been shown to be crucially involved in regulation of barrier properties. By laser tweezer application we demonstrated that the strength of adhesion of VE-cadherin-coated microspheres to the surface of cultured endothelial monolayers is significantly reduced by treatment with two well-established permeability-increasing compounds, cytochalasin D and the Ca(2+) ionophore A23187, which shows that both compounds directly affect cadherin mediated adhesion. Cytochalasin D and A23187 caused considerable decay of F-actin (30-60%). Stabilisation of F-actin by jasplakinolide completely blocked drug induced weakening of bead adhesion showing that attenuation of cadherin-cadherin trans-interaction induced by cytochalasin D and A23187 depends largely on downregulation of F-actin. Single molecule fluorescence microscopy demonstrated that drug-induced weakening of adhesion is accompanied by an increase in lateral mobility of cadherins as well as by dispersal of cadherin-enriched plasmalemmal microdomains. However, the lifetime ( approximately 700 milliseconds, k(off) approximately 1.4 second(-1)) and apparent on-rate of cadherin trans-interaction (relative frequency of binding) remained unchanged in response to cytochalasin D and A23187 indicating that cadherin-mediated adhesion is not modulated by inside out changes of the affinity but, rather, appears to be controlled by actin dependent tethering and compartmentalization of cadherins. PMID- 12584246 TI - Deletion of proteasomal subunit S5a/Rpn10/p54 causes lethality, multiple mitotic defects and overexpression of proteasomal genes in Drosophila melanogaster. AB - The regulatory complex of the 26S proteasome is responsible for the selective recognition and binding of multiubiquitinated proteins. It was earlier shown that the subunit S5a/Rpn10/p54 of the regulatory complex is the only cellular protein capable of binding multiubiquitin chains in an in vitro overlay assay. The role of this subunit in substrate selection, however, is a subject of debate, following the observation that its deletion in Saccharomyces cerevisiae is not lethal and instead causes only a mild phenotype. To study the function of this subunit in higher eukaryotes, a mutant Drosophila strain was constructed by deleting the single copy gene encoding subunit S5a/Rpn10/p54. This deletion caused larval-pupal polyphasic lethality, multiple mitotic defects, the accumulation of higher multimers of ubiquitinated proteins and a huge accumulation of defective 26S proteasome particles. Deletion of the subunit S5a/Rpn10/p54 does not destabilise the regulatory complex and does not disturb the assembly of the regulatory complex and the catalytic core. The pupal lethality is a consequence of the depletion of the maternally provided 26S proteasome during the larval stages and a sudden increase in the proteasomal activity demands during the first few hours of pupal development. The huge accumulation of the fully assembled 26S proteasome in the deletion mutant and the lack of free subunits or partially assembled particles indicate that there is a highly coordinated accumulation of all the subunits of the 26S proteasome. This suggests that in higher eukaryotes, as with yeast, a feedback circuit coordinately regulates the expression of the proteasomal genes, and this adjusts the actual proteasome concentration in the cells according to the temporal and/or spatial proteolytic demands. PMID- 12584245 TI - Tumour-endothelium interactions in co-culture: coordinated changes of gene expression profiles and phenotypic properties of endothelial cells. AB - Tumour angiogenesis is a complex process based upon a sequence of interactions between tumour cells and endothelial cells. To model tumour/endothelial-cell interactions, we co-cultured U87 human glioma cells with human umbilical vein endothelial cells (HUVECs). U87 cells induced an 'activated' phenotype in HUVECs, including an increase in proliferation, migration and net-like formation. Activation was observed in co-cultures where cells were in direct contact and physically separated, suggesting an important role for soluble factor(s) in the phenotypic and genotypic changes observed. Expressional profiling of tumour activated endothelial cells was evaluated using cDNA arrays and confirmed by quantitative PCR. Matching pairs of receptors/ligands were found to be coordinately expressed, including TGFbetaRII with TGFbeta3, FGFRII and cysteine rich fibroblast growth factor receptor (CRF-1) with FGF7 and FGF12, CCR1, CCR3, CCR5 with RANTES and calcitronin receptor-like gene (CALCRL) with adrenomedullin. Consistent with cDNA array data, immunohistochemical staining of expressed proteins revealed the upregulation of Tie-2 receptor in vitro and in vivo. Our data suggest that tumour-induced activation of quiescent endothelial cells involves the expression of angiogenesis-related receptors and the induction of autocrine growth loops. We suggest that tumour cells release growth factors that induce endothelial cells to express specific ligands and their cognate receptors coordinately. PMID- 12584247 TI - Abnormal tissue-dependent polytenization of a block of chromosome 3 pericentric heterochromatin in Drosophila melanogaster. AB - Heterochromatic DNA sequences in the polytene chromosomes of Drosophila melanogaster salivary glands are under-replicated in wild-type strains. In salivary glands of SuUR and in the nurse cells of otu mutants, under-replication is partly suppressed and a banded structure appears within the centric heterochromatin of chromosome 3. This novel banded structure in salivary gland chromosomes was called Plato Atlantis. In order to characterize the heterochromatic component of Plato Atlantis, we constructed a fine-scale cytogenetic map of deletions with break points within centric heterochromatin (Df(3L)1-16, Df(3L)2-66, Df(3R)10-65, Df(3R)4-75 and Df(3L)6B-29 + Df(3R)6B-29). Salivary gland chromosomes show that Df(3L)1-16 removes the complete Plato Atlantis, while Df(3L)2-66 deletes the most proximal 3L regions. These deletions therefore show a substantial cytological overlap. However, in the chromosomes of nurse cells, the same deficiencies remove distinct heterochromatic blocks, with the region of overlap being almost invisible. Satellite (AATAACATAG, AAGAG) and dodecasatellite DNAs mapped in a narrow interval in salivary glands but were found in three clearly distinguishable blocks in nurse cells. The 1.688 satellite was found at a single site in salivary glands but at two sites in nurse cells. We show that newly polytenized heterochromatic structures include blocks h47-h50d of mitotic heterochromatin in salivary glands, but the additional blocks h50p, h53 and h57 are also included in nurse cell chromosomes. Tissue specificity of the patterns of abnormal heterochromatic polytenization implies differential control of DNA replication in somatic and germline cells. PMID- 12584248 TI - Protein products of human Gas2-related genes on chromosomes 17 and 22 (hGAR17 and hGAR22) associate with both microfilaments and microtubules. AB - The human Gas2-related gene on chromosome 22 (hGAR22) encodes two alternatively spliced mRNA species. The longer mRNA encodes a protein with a deduced molecular mass of 36.3 kDa (GAR22alpha), whereas the shorter mRNA encodes a larger protein with a deduced molecular mass of 72.6 kDa (GAR22beta). We show that both hGAR22 proteins contain a calponin homology actin-binding domain and a Gas2-related microtubule-binding domain. Using rapid amplification of cDNA ends, we have cloned the mouse orthologue of hGAR22, mGAR22, and found its protein products to be extremely well conserved. We also report the cDNA cloning of a human Gas2 related gene on chromosome 17 (hGAR17). hGAR17 also encodes two protein isoforms. The overall cytoskeletal binding properties of the hGAR17 and hGAR22 proteins are remarkably similar. hGAR17 mRNA expression is limited to skeletal muscle. Although hGAR22 and mGAR22 mRNAs are expressed nearly ubiquitously, mGAR22 protein can only be detected in testis and brain. Furthermore, only the beta isoform is present in these tissues. GAR22beta expression is induced in a variety of cultured cells by growth arrest. The absolute amounts of GAR22beta protein expressed are low. The beta isoforms of hGAR17 and hGAR22 appear to be able to crosslink microtubules and microfilaments in transfected cells. This finding suggests that the physiological functions of these proteins may involve integration of these two components of the cytoskeleton. PMID- 12584249 TI - Distinct caveolae-mediated endocytic pathways target the Golgi apparatus and the endoplasmic reticulum. AB - Internalization of autocrine motility factor (AMF) into the endoplasmic reticulum is sensitive to the cholesterol-extracting reagent methyl-beta-cyclodextrin, inhibited by the dynamin-1 K44A mutant and negatively regulated by caveolin-1. Thus, AMF internalization requires a caveolae-mediated endocytic pathway. Similarly, we show here that endocytosis of cholera toxin (CTX) in NIH-3T3 fibroblasts is inhibited by adenoviral expression of the dynamin-1 K44A mutant but only partially by expression of the clathrin hub. Treatment with methyl-beta cyclodextrin and overexpression of caveolin-1, but not the clathrin hub, selectively diminishes CTX endocytosis to the Golgi apparatus but not to endosomes. CTX is therefore targeted via a caveolin-1-regulated caveolae-mediated pathway to the Golgi. Disruption of Golgi-, caveosome- or endosome-mediated trafficking with brefeldin A, nocodazole or a 20 degrees C temperature block, respectively, inhibit CTX endocytosis to the Golgi but do not affect AMF delivery to the endoplasmic reticulum. Following an incubation of only five minutes in the presence of the clathrin hub, AMF and CTX are not cointernalized, and AMF is delivered to the AMF-R-positive smooth ER. The internalization of both ligands is nevertheless sensitive to the tyrosine kinase inhibitor genistein, confirming that they are both internalized via caveolae/raft pathways. Two distinct caveolae mediated endocytic pathways therefore exist, including a novel direct pathway to the ER from the plasma membrane. PMID- 12584250 TI - Reduced survival of lens epithelial cells in the alphaA-crystallin-knockout mouse. AB - alphaA-Crystallin (alphaA) is a molecular chaperone expressed preferentially in the lens. alphaA transcripts are first detected during the early stages of lens development and its synthesis continues as the lens grows throughout life. alphaA(-/-) mouse lenses are smaller than controls, and lens epithelial cells derived from these mice have diminished growth in culture. In the current work, we tested the hypothesis thatalphaA prevents cell death at a specific stage of the cell cycle in vivo. Seven-day-old 129Sv (wild-type) and alphaA(-/-) mice were injected with 5-bromo-2'-deoxyuridine (BrdU) to label newly synthesized DNA in proliferating cells. To follow the fate of the labeled cells, wholemounts of the capsule epithelial explants were made at successive times after the BrdU pulse, and the labeling index was determined. Immunofluorescence and confocal microscopy showed that both wild-type and alphaA(-/-) cells had a 3-hour labeling index of 4.5% in the central region of the wholemount, indicating that the number of cells in S phase was the same. Twenty-four hours after the pulse, individual cells labeled with BrdU had divided and BrdU-labeled cells were detected in pairs. The 24-hour labeling index in the wild-type lens was 8.6%, but in the alphaA(-/-) lens it was significantly lower, suggesting that some of the cells failed to divide and/or that the daughter cells died during mitosis. TUNEL labeling was rarely detected in the wild-type lens, but was significant and always detected in pairs in the alphaA(-/-) wholemounts. Dual labeling with TUNEL and BrdU also suggested that the labeled cells were dying in pairs in the alphaA(-/-) lens epithelium. Immunolabeling of wholemounts with beta-tubulin antibodies indicated that the anaphase spindle in a significant proportion of alphaA(-/-) cells was not well organized. Examination of the cellular distribution of alphaA in cultured lens epithelial cells showed that it was concentrated in the intercellular microtubules of cells undergoing cytokinesis. These data suggest that alphaA expression in vivo protects against cell death during mitosis in the lens epithelium, and the smaller size of the alphaA(-/-) lens may be due to a decrease in the net production of epithelial cells. PMID- 12584251 TI - Retrograde flow rate is increased in growth cones from myosin IIB knockout mice. AB - Growth cones of myosin-IIB-knockout mice have reduced outgrowth rates and traction force. There is a close relationship between traction force, retrograde flow and forward advance of growth cones. All three activities appear to be at least partially myosin dependent. Therefore, we have now tested for differences in retrograde flow rates between growth cones from myosin-IIB-knockout mice and their normal littermates. By placing nerve-growth-factor-coated silica beads on the surface of growth cones with laser tweezers, or by tracking GFP-myosin IIA spots, we found that the retrograde flow rate was increased more than two fold in the knockout growth cones compared with the wild type. These data suggest that both myosin IIA and IIB normally contribute to retrograde flow and the properties of the flow are strongly influenced by myosin IIB because of its location and abundance. However, in the absence of myosin IIB, myosin IIA takes over this function. The change in retrograde flow rate may reflect the difference in functional properties of these two myosins. Knockout growth cones also exhibited reduced stability of lamellipodia, possibly as a partial consequence of this increased retrograde flow rate. In addition, microtubules penetrated a shorter distance into filopodia, which suggests that the increase in flow rate may adversely affect the microtubule-dependent maturation of filopodia. Taken together these data support the idea that the forward advance of the growth cone is myosin II dependent and involves multiple myosin II isoforms. PMID- 12584252 TI - Spatial and cellular localization of calcium-dependent protease (CDP II) in Allomyces arbuscula. AB - Immunogold labeling of calcium-dependent neutral protease II (CDPII) with specific antibodies in near median longitudinal ultrathin sections of Allomyces arbuscula showed that the enzyme is predominantly localized in the growing hyphal and rhizoidal apices. The tips in both cell type had more enzyme than the distal regions and showed a gradient distribution. Labeling of the ultrathin sections and western blot analysis of purified subcellular fractions showed that CDPII is mainly cytosolic. Catalytic activity of the enzyme measured with synthetic substrate (Bz-Arg-pNA) showed that 90% of its activity is present in the soluble fraction, although a small amount is associated with the nuclei (0.2%), plasma membranes (0.7%) and microsomes (3.9%). This association is discussed in the context of the functional role of the enzyme and its possible localized activation. Western blot analysis of the crude extract and indirect immunofluorescence of the fixed permeabilized hypahe after treatment with CDPII showed that the alpha-tubulin is a specific target of the enzyme. PMID- 12584253 TI - Role of the Vtc proteins in V-ATPase stability and membrane trafficking. AB - Vtc proteins have genetic and physical relations with the vacuolar H(+)-ATPase (V ATPase), influence vacuolar H(+) uptake and, like the V-ATPase V(0) sectors, are important factors in vacuolar membrane fusion. Vacuoles from vtc1delta and vtc4delta mutants had slightly reduced H(+)-uptake activity. These defects could be separated from Vtc function in vacuole fusion, demonstrating that Vtc proteins have a direct role in membrane fusion. We analyzed their involvement in other membrane trafficking steps and in VATPase dynamics. Deletion of VTC genes did not impede endocytic trafficking to the vacuole. However, ER to Golgi trafficking and further transport to the vacuole was delayed in deltavtc3 cells. In accordance with that, deltavtc3 cells showed a reduced growth rate. Vtc mutations did not interfere with regulated assembly and disassembly of the V-ATPase, but they affected the number of peripheral V(1) subunits associated with the vacuoles. deltavtc3 vacuoles carried significantly more V(1) subunits, whereas deltavtc1, deltavtc2 and deltavtc4 had significantly less. The proteolytic sensitivity of the V(0) subunit Vph1p was different in deltavtc and wild-type cells in vivo, corroborating the physical interaction of Vtc proteins with the V-ATPase observed in vitro. We suggest that Vtc proteins affect the conformation of V(0). They might thereby influence the stability of the VATPase holoenzyme and support the function of its V(0) sector in vacuolar membrane fusion. PMID- 12584254 TI - In situ visualization of rDNA arrangement and its relationship with subnucleolar structural regions in Allium sativum cell nucleolus. AB - We used a DNA-specific staining technique to show the two states of DNA component distributed in the nucleolar region of Allium sativum cells. One state is the extended DNA fiber, and the other is the condensed DNA clump. In situ hybridization demonstrated that the extended DNA fiber was an rRNA gene. Anti fibrillarin antibody immunolabeling revealed that these rRNA genes were located in the dense fibrillar component near the fibrillar center, including at the periphery of the fibrillar center. None was in the dense fibrillar component far away from the fibrillar center. The condensed DNA clump was located in the fibrillar center. Further observations showed that the rRNA genes in the nucleolus were all arranged around the fibrillar center and associated with the DNA clumps in the fibrillar center. Results of statistical analysis showed that the distribution region of rRNA genes occupied about one-third of the total dense fibrillar component region. Ag-NOR protein showed a similar distribution pattern to that of rDNA. Immunolabeling of an anti-RNA/DNA hybrid antibody demonstrated that the transcription sites of rRNA were located at the periphery of the fibrillar center and in the dense fibrillar component near the fibrillar center, and these sites were consistent with the location and arrangement of rDNA shown in situ. These results demonstrated that transcription of rRNA takes place around the fibrillar center and at the periphery, whereas the dense fibrillar component that was far away from fibrillar center was the non-transcription region. The DNA clumps within the fibrillar center were probably the anchoring sites for rDNA arrangement. PMID- 12584255 TI - gamma-Secretase activity requires the presenilin-dependent trafficking of nicastrin through the Golgi apparatus but not its complex glycosylation. AB - Nicastrin and presenilin are two major components of the gamma-secretase complex, which executes the intramembrane proteolysis of type I integral membrane proteins such as the amyloid precursor protein (APP) and Notch. Nicastrin is synthesized in fibroblasts and neurons as an endoglycosidase-H-sensitive glycosylated precursor protein (immature nicastrin) and is then modified by complex glycosylation in the Golgi apparatus and by sialylation in the trans-Golgi network (mature nicastrin). These modifications are not observed with exogenously overexpressed nicastrin. Under normal cell culture conditions, only mature nicastrin is expressed at the cell surface and binds to the presenilin heterodimers. Mature nicastrin has a half-life of more than 24 hours. In the absence of presenilin 1 and 2, nicastrin remains entirely endoglycosidase H sensitive, is retained in the endoplasmic reticulum and is slowly degraded. Single presenilin 1 or presenilin 2 deficiency affects glycosylation of nicastrin to a lesser extent than the combined presenilin deficiencies, suggesting a correlation between either the transport of nicastrin out of the endoplasmic reticulum or the concomitant complex glycosylation of nicastrin, and gamma secretase activity. However, when complex glycosylation of nicastrin was inhibited using mannosidase I inhibitors, gamma-secretase cleavage of APP or Notch was not inhibited and the immature nicastrin still associates with presenilin and appears at the cell surface. Complex glycosylation of nicastrin is therefore not needed for gamma-secretase activity. Because the trafficking of nicastrin to the Golgi apparatus is dependent on presenilins, our data point to a central role of presenilin in nicastrin maturation/localization, which could help to partially resolve the 'spatial paradox'. PMID- 12584256 TI - A beta-catenin survival signal is required for normal lobular development in the mammary gland. AB - The Wnt (wingless) family of secreted glycoproteins initiates a signalling pathway implicated in the regulation of both normal mouse mammary gland development and tumorigenesis. Multiple Wnt signals ultimately converge on the multifunctional protein beta-catenin to activate the transcription of target genes. Although beta-catenin plays a crucial role in canonical Wnt signalling, it also functions in epithelial cell-cell adhesion at the adherens junctions. This study was designed to isolate beta-catenin's signalling function from its role in adherence during mouse mammary gland development. A transgenic dominant-negative beta-catenin chimera (beta-eng), which retains normal protein-binding properties of wild-type beta-catenin but lacks its C-terminal signalling domain, was expressed preferentially in the mammary gland. Thus, beta-eng inhibits the signalling capacity of endogenous beta-catenin, while preserving normal cell-cell adhesion properties. Analysis of the mammary gland in transgenic mice revealed a severe inhibition of lobuloalveolar development and a failure of the mice to nurse their young. Expression of beta-eng resulted in an induction of apoptosis both in transgenic mice and in retrovirally transduced HC11 cells. Thus, endogenous beta-catenin expression appears to be required to provide a survival signal in mammary epithelial cells, which can be suppressed by transgenic expression of beta-eng. Comparison of the timing of transgene expression with the transgenic phenotype suggested a model in which beta-catenin's survival signal is required in lobular progenitors that later differentiate into lobuloalveolar clusters. This study illustrates the importance of beta-catenin signalling in mammary lobuloalveolar development. PMID- 12584259 TI - Mechanisms of oedema in nephrotic syndrome: old theories and new ideas. PMID- 12584260 TI - Monocyte chemoattractant protein-1: does it play a role in diabetic nephropathy? PMID- 12584261 TI - Hyperglycaemia and renal ischaemia-reperfusion injury. PMID- 12584262 TI - Uraemic toxins and cardiovascular disease. PMID- 12584263 TI - Dendritic cells and the mode of action of anticalcineurinic drugs: an integrating hypothesis. PMID- 12584265 TI - Age and renal transplantation: an interim analysis. PMID- 12584266 TI - Third International Meeting on Cyclooxygenase-2--basic research and therapeutic use, Regensburg, Germany, June 7-8, 2002. PMID- 12584267 TI - Increase in nitric oxide bioavailability improves endothelial function in endothelin-1 transgenic mice. AB - BACKGROUND: Endothelin-1 (ET-1) has been described as a very potent vasoconstrictor. Nevertheless, transgenic mice overexpressing ET-1 have been shown to exhibit normal blood pressure. We thus hypothesized that vascular ET-1 effects may be antagonized by increased activity of other regulatory systems, such as the increase in bioavailability of the endothelial counterpart of ET-1, nitric oxide (NO). METHODS: Endothelium-dependent and -independent vascular function was assessed as relaxation/contraction of isolated preconstricted aortic rings to acetylcholine (10(-10)-10(-4) mol/l), sodium nitroprusside (10(-10)-10( 4) mol/l), ET-1 (10(-10)-10(-7) mol/l) and big ET-1 (10(-10)-10(-7) mol/l), respectively, in ET-1 transgenic mice and corresponding controls. To unmask the impact of the NO system, we furthermore analysed vessel rings incubated in vitro with the NO-synthase inhibitor L-N(G)-nitroarginine methyl ester (L-NAME, 10(-4) mol/l). RESULTS: Maximum endothelium-dependent relaxation was enhanced in ET-1 transgenic mice (93+/-3% vs 84+/-4% for wild-type littermates; P<0.05) and was inhibited by preincubation with L-NAME in both ET-transgenic mice and wild-type littermates (11+/-5% vs 9+/-4% maximum relaxation, respectively). Endothelium independent relaxation was similar among all groups. Maximum vascular contraction to ET-1 and big ET-1 was reduced in ET-1 transgenic mice (P<0.05 vs wild-type littermates). Preincubation with L-NAME reduced this difference, indicating the involvement of augmented NO availability. Correspondingly, urinary nitrate/nitrite excretion was significantly elevated in ET-1 transgenic mice. CONCLUSIONS: These data suggest that in transgenic mice overexpressing ET-1, increased NO bioavailability counteracts the contractile potency of elevated ET-1 levels and leads to an improvement of endothelium-dependent relaxation. Thus, in the presence of an activated ET system, up-regulation of NO production may be capable of maintaining vascular tone in a normal range and therefore may prevent the development of hypertension. PMID- 12584268 TI - Suppressive effects of Perilla frutescens on IgA nephropathy in HIGA mice. AB - BACKGROUND: Perilla frutescens (perilla) is a herbal medicine used in Japanese traditional Kampo medicine. The present study was conducted to evaluate the anti nephritic effects of perilla in HIGA mice that spontaneously develop high levels of serum immunoglobulin A (IgA) along with mesangial IgA deposition. METHODS: A perilla decoction and its major active constituent, rosmarinic acid (RsA), were orally administrated to 10-week-old HIGA mice for 16 weeks. At study completion, we measured proteinuria and serum IgA levels and generated histological scores from kidney specimens. In addition, we measured concentrations of IgA in culture media of intestinal Peyer's patch cells and spleen cells obtained from the HIGA mice. RESULTS: Perilla suppressed proteinuria, proliferation of glomerular cells, serum levels of IgA, glomerular IgA and IgG depositions in HIGA mice. Cultured Peyer's patch cells and spleen cells from perilla-treated mice produced significantly less IgA than controls. Rosmarinic acid, by itself, suppressed serum IgA levels and glomerular IgA deposition in HIGA mice. Cultured spleen cells from RsA-treated mice produced less IgA than controls. CONCLUSIONS: The perilla decoction may suppress IgA nephropathy, in part, through modulation of the intestinal mucosal immune system. These effects were caused by RsA acting synergistically with other constituents. PMID- 12584269 TI - Kidney function and morphology after short-term combination therapy with cyclosporine A, tacrolimus and sirolimus in the rat. AB - BACKGROUND: Sirolimus (SRL) may supplement calcineurin inhibitors in clinical organ transplantation. These are nephrotoxic, but SRL seems to act differently displaying only minor nephrotoxic effects, although this question is still open. In a number of treatment protocols where SRL was combined with a calcineurin inhibitor indications of a synergistic nephrotoxic effect were described. The aim of this study was to examine further the renal function, including morphological analysis of the kidneys of male Sprague-Dawley rats treated with either cyclosporine A (CsA), tacrolimus (FK506) or SRL as monotherapies or in different combinations. METHODS: For a period of 2 weeks, CsA 15 mg/kg/day (given orally), FK506 3.0 mg/kg/day (given orally) or SRL 0.4 mg/kg/day (given intraperitoneally) was administered once a day as these doses have earlier been found to achieve a significant immunosuppressive effect in Sprague-Dawley rats. In the 'conscious catheterized rat' model, the glomerular filtration rate (GFR) was measured as the clearance of Cr(EDTA). The morphological analysis of the kidneys included a semi quantitative scoring system analysing the degree of striped fibrosis, subcapsular fibrosis and the number of basophilic tubules, plus an additional stereological analysis of the total grade of fibrosis in the cortex stained with Sirius Red. RESULTS: CsA, FK506 and SRL all significantly decreased the GFR. A further deterioration was seen when CsA was combined with either FK506 or SRL, whereas the GFR remained unchanged in the group treated with FK506 plus SRL when compared with treatment with any of the single substances. The morphological changes presented a similar pattern. The semi-quantitative scoring was significantly worst in the group treated with CsA plus SRL (P<0.001 compared with controls) and the analysis of the total grade of fibrosis also showed the highest proportion in the same group and was significantly different from controls (P<0.02). The FK506 plus SRL combination showed only a marginally higher degree of fibrosis as compared with controls (P=0.05). CONCLUSION: This rat study demonstrated a synergistic nephrotoxic effect of CsA plus SRL, whereas FK506 plus SRL was better tolerated. PMID- 12584270 TI - Vasopressin increases urinary albumin excretion in rats and humans: involvement of V2 receptors and the renin-angiotensin system. AB - BACKGROUND: An increase in urinary albumin excretion (UAE) represents an early predictor of glomerular damage in diabetes mellitus (DM) and a risk factor for cardiovascular complications in hypertension. Vasopressin is elevated in DM and in some forms of hypertension. Previous studies in rats suggested that this hormone could play a role in the albuminuria observed in chronic renal failure or diabetic nephropathy, but no information is available concerning the mechanism of these effects and the possible influence of vasopressin on UAE in the healthy kidney. The present study was thus designed to evaluate whether vasopressin influences UAE in normal rats and humans, whether this effect is V(2)-receptor dependent, and whether it is mediated by the renin-angiotensin system. METHODS: UAE was measured in normal Wistar rats and healthy humans, or in subjects with various forms of diabetes insipidus (DI), before and after acute or chronic infusion of the vasopressin V(2) receptor agonist dDAVP. Chronic dDAVP administration was also performed in normal Wistar rats previously submitted to either chronic angiotensin-converting enzyme inhibition (ACEI) or chronic blockade of AT1 receptors (ARB). RESULTS: In rats, acute or chronic dDAVP infusion increased UAE significantly and reversibly (4-fold and 6-fold, respectively). In healthy subjects, acute infusion of dDAVP tripled UAE (P<0.01) but did not change creatinine and beta(2)-microglobulin excretion, thus suggesting that the rise in UAE was due to an increased glomerular leakage of albumin. dDAVP also increased UAE in patients with central DI and in patients with hereditary nephrogenic DI bearing AQP2 mutations. However, UAE was not increased in patients with hereditary nephrogenic DI bearing mutations of the V(2) receptor. In rats, ACEI and ARB blunted the dDAVP-induced rise in UAE by 70% (P<0.05) and 50% (NS), respectively. CONCLUSIONS: The present studies reveal for the first time that vasopressin induces a marked increase in UAE in healthy rats and humans. This albuminuric effect seems to result from increased glomerular leakage, requires functional vasopressin V(2) receptors, and is, at least in part, mediated by the renin-angiotensin system. These results bring additional support for an involvement of vasopressin in the albuminuria observed in pathological states such as diabetes mellitus or hypertension. PMID- 12584271 TI - Urinary calcium excretion in children with vesicoureteral reflux. AB - BACKGROUND: Renal malformations including vesico-ureteral reflux (VUR) are associated with urolithiasis. However, studies on urinary calcium excretion in children with VUR have not been reported. This study was conducted to find out whether children with VUR have a higher prevalence of hypercalciuria and whether their family members are affected by hypercalciuria and/or urolithiasis. METHODS: We studied the prevalence of hypercalciuria and urolithiasis in 46 children (12 males and 34 females) with VUR and in their parents. RESULTS: Three out of 46 children had renal colic and nine out of 46 exhibited calyceal microlithiasis in the renal sonography. According to Stapleton's criteria, we found that 27 out of 46 children (58.6%) had hypercalciuria. These children were significantly shorter than children with normal calciuria and showed lower values of maximal urinary osmolality. We found no differences in urinary calcium excretion values related to the VUR grading, or to the presence or absence of renal scars, or to whether VUR was still unresolved or already resolved at the time of study. Seventeen out of 27 children with hypercalciuria (63%) had one or both parents affected by hypercalciuria, and there was a history of urolithiasis in six first-degree relatives and in four second-degree relatives (37%). Besides, 10 out of 19 children without hypercalciuria (52.6%) had one or both parents affected by hypercalciuria and there was a history of urolithiasis in three first-degree relatives and in three second-degree relatives (31.6%). Among the 27 children whose parents had hypercalciuria, four had both parents affected, 19 had only the mother affected and in four patients only the father was affected. CONCLUSION: Our results showed that the prevalence of hypercalciuria was greater in paediatric patients with VUR than in the general population. Urolithiasis in patients with VUR had a metabolic origin. Hypercalciuria was inherited as an autosomal dominant trait although with a higher probability to be inherited from the mother. PMID- 12584272 TI - Exclusion of mutations in FXYD2, CLDN16 and SLC12A3 in two families with primary renal Mg2+ loss. AB - BACKGROUND: Based on genetic studies in families with hereditary renal Mg(2+) reabsorption disorders, several genes were shown to be involved in renal Mg(2+) transport. Mutations in the CLDN16 gene were found to underlie autosomal recessive hypomagnesaemia associated with hypercalciuria and nephrocalcinosis. The FXYD2 gene was implicated in autosomal dominant renal Mg(2+) wasting associated with hypocalciuria. Mutations in the SLC12A3 gene, also known as NCC, cause Gitelman's syndrome. In addition to hypokalaemic metabolic alkalosis, hypomagnesaemia associated with hypocalciuria is considered to be a hallmark feature of this latter disorder. METHODS: We have characterized a new family with presumed dominant renal hypomagnesaemia by detailed clinical examination and mutation analysis of CLDN16, FXYD2 and SLC12A3. In addition, we have performed mutation analysis of these three genes in a previously described family with autosomal recessive renal Mg(2+) wasting. In this family, linkage analysis was performed with polymorphic markers in the vicinity of the FXYD2 gene. RESULTS: The phenotype of the new family closely resembles that of the known dominant families with a mutation in FXYD2, but mutations in this gene were not identified in the new family. No mutations were found in CLDN16 and SLC12A3 either. Sequencing of the three genes in the patients of the recessive family revealed no mutations. In addition, haplotype analysis excluded linkage to the FXYD2 region on chromosome 11q23. CONCLUSION: Our results indicate that, in addition to the currently known loci involved in renal Mg(2+) handling, at least one other gene must be involved. PMID- 12584273 TI - Clinical prognostic factors in biopsy-proven benign nephrosclerosis. AB - BACKGROUND: Hypertensive renal damage has become one of the most important causes of end-stage renal failure (ESRF) in Western countries. Affected patients rarely have a kidney biopsy and their diagnoses therefore remain uncertain. The objective of the present study was to examine patients suspected of renal glomerular disease, which at biopsy proved to have isolated benign nephrosclerosis. We wanted to study the effect of different clinical and laboratory variables at the time of biopsy on the short-term and long-term progression to ESRF and death. METHODS: We retrospectively examined 102 patients who were diagnosed by kidney biopsy in Norway between April 1988 and December 1990. All patients were followed by means of registries for approximately 13 years to describe renal and patient survival. RESULTS: The age of the patients at the time of biopsy was 55+/-15 years (range 15-88 years). Three years after the time of biopsy, 18% had developed ESRF and 24% had died; the corresponding numbers 13 years after biopsy were 32% and 47%. By Kaplan-Meier analyses, the following variables indicated short-term progression to ESRF: serum creatinine > or = 200 micro mol/l, systolic blood pressure > or = 160 mmHg and proteinuria > or = 1 g/24 h. In addition, patients with increased diastolic blood pressure, increased age and decreased serum albumin tended to develop ESRF more often. Long term predictors of ESRF in Kaplan-Meier analyses were increased serum creatinine and urinary protein. Independent risk factors for progression to ESRF were increased serum creatinine and increased urinary protein. Independent risk factors for death were increased age and increased serum creatinine. CONCLUSIONS: Benign nephrosclerosis is a common condition that is associated with a high morbidity and mortality. Short-term predictors of ESRF differ from long-term predictors and this may reflect a pathophysiologically meaningful difference. PMID- 12584275 TI - Familial ATTR amyloidosis: microalbuminuria as a predictor of symptomatic disease and clinical nephropathy. AB - BACKGROUND: Portuguese type familial amyloid polyneuropathy (FAP) is a neuropathic amyloidosis caused by a mutant transthyretin (TTR). Varying degrees of renal involvement have been reported. Our aim was to assess the value of microalbuminuria (MA) for predicting clinical neurological disease and overt nephropathy in TTR-related amyloidosis. METHODS: All subjects had the TTR Val30Met mutation, and were recruited between 1993 and 1999. We have prospectively evaluated 22 asymptomatic gene carriers (7 male, 15 female; mean age 41.6+/-9.6 years) and 32 patients with neuropathy (14 male, 18 female; 36.8+/ 8.8 years, on average, 33.0+/-9.3 years at the onset of neuropathy). We measured urinary albumin excretion every year, if asymptomatic, or every 6 months if already affected. Kidney biopsies were performed in patients with normal urinary albumin excretion, MA, and overt nephropathy, respectively. RESULTS: In asymptomatic carriers, persistent MA was detected in eight (36%) subjects. The presence of MA in asymptomatic gene carriers, compared with those having normal urinary albumin excretion, conferred a 4.8-fold risk of developing neuropathy, usually within the subsequent 3 years. Once neurological signs appeared, nephropathy, manifested as MA, progressed to overt nephropathy in one-half of subjects. In patients with neuropathy, 24 (75%) had MA during follow-up: evolution towards clinical renal disease occurred in 14 (58%) and renal failure occurred in five (21%), always after a course of MA. Proteinuria or renal failure without prior persistent MA were never observed in the present patient cohort. Histopathological evaluation did not reveal glomerular lesions other than amyloid deposits to explain abnormal urinary albumin excretion. The amount of mesangial and vascular-pole amyloid deposits was correlated with the degree of albuminuria. CONCLUSIONS: Microalbuminuria represents the first stage of clinical TTR amyloid nephropathy and is premonitory of neuropathy. Its presence identifies a subgroup of patients who are more prone to develop overt nephropathy. Screening of MA may be important to assess disease onset and to recommend liver transplantation in individuals at risk. PMID- 12584274 TI - Glomerular immune deposits are associated with increased proteinuria in patients with ANCA-associated crescentic nephritis. AB - BACKGROUND: In small vessel vasculitis and its renal-limited form, idiopathic crescentic glomerulonephritis, renal damage is characterized by pauci-immune necrotizing crescentic glomerulonephritis (CGN) without histological evidence of immunoglobulin (Ig) deposition. In some patients, however, significant amounts of immune deposits may be detected. Therefore, we evaluated the clinical significance of these immune deposits in anti-neutrophil cytoplasmic autoantibody (ANCA)-associated pauci-immune CGN. METHODS: Renal biopsies of 45 consecutive patients with new onset of Wegener's granulomatosis, microscopic polyangiitis and idiopathic CGN were retrospectively evaluated by light microscopy, immunohistochemistry and electron microscopy and the findings compared with renal function and outcome. RESULTS: Typical pauci-immune CGN was found in 37 patients (group I). In eight patients (18%; group II), however, histopathological examination revealed substantial deposition of Ig in the mesangium and/or along the glomerular basement membrane. Five of these eight patients were cANCA positive; two initially had pANCA and developed a cANCA pattern and one was pANCA positive. There were no differences between groups in age, gender, renal function or extra-renal organ involvement at the time of biopsy. However, patients in group II had significantly more proteinuria (5.4+/-3.1 vs 1.3+/-1.0 g/24 h; P=0.016). We also observed a trend for a worse outcome with respect to renal function and mortality in group II patients; however, the differences did not reach significance. CONCLUSIONS: Our results confirm that in ANCA-associated CGN a substantial percentage of patients have evidence of Ig deposition in renal biopsies. In this subgroup, Ig deposition was associated with a significantly greater degree of proteinuria. Further investigations are necessary to define the full clinical impact of immune-complex deposition on the clinical course of renal disease in pauci-immune CGN. PMID- 12584276 TI - Impact of low-dose acetylsalicylic acid on kidney function in type 2 diabetic patients with elevated urinary albumin excretion rate. AB - BACKGROUND: Low-dose treatment with acetylsalicylic acid (ASA) is widely recommended to type 2 diabetic patients as primary prevention against cardiovascular disease. High-dose treatment with cyclooxygenase inhibitors reduces urinary albumin excretion rate (AER) in type 1 diabetic patients with micro- or macroalbuminuria. Whether a similar effect on AER exists during low dose ASA treatment, which may confound the diagnosis and monitoring of micro- and macroalbuminuria in type 2 diabetic patients, remains to be elucidated. METHODS: In a randomized, double-blind, crossover trial, 31 type 2 diabetic patients with elevated levels of AER (>30 mg/24 h) were, in random order, given ASA (150 mg/day) for 4 weeks followed by placebo for 4 weeks with a 2 week washout period or vice versa. At the end of each treatment period AER, glomerular filtration rate (GFR), blood pressure (BP), transcapillary escape rate (TER(alb)) of albumin and haemoglobin A(1c) (HbA(1c)) were measured. RESULTS: The following variables remained unchanged (mean (95% CI) unless otherwise noted) (ASA vs placebo, paired Student's t-test): AER (201 (119-341) vs 205 (124-340) mg/24 h (geometric mean, 95% CI); P=0.78), GFR (103 (94-111) vs 102 (93-110) ml/min; P=0.58), systolic BP (151 (146-158) vs 152 (146-158) mmHg; P=0.68), diastolic BP (87 (83-91) vs 87 (82 91) mmHg; P=0.88), TER(alb) (6.3 (5.7-6.9) vs 5.9 (5.1-6.7); P=0.45) and HbA(1c) (8.6 (8.1-9.0) vs 8.5 (8.1-9.0) %; P=0.60). CONCLUSIONS: Low-dose treatment with 150 mg ASA daily does not have any impact on AER or GFR in type 2 diabetic patients with micro- or macroalbuminuria. Consequently, the widely recommended prescription of low-dose ASA as a primary and secondary prevention strategy against cardiovascular disease in these patients does not confound the diagnosis or monitoring of micro- or macroalbuminuria. PMID- 12584277 TI - Measurement of tubular enzymuria facilitates early detection of acute renal impairment in the intensive care unit. AB - BACKGROUND: Early detection of acute tubular necrosis (ATN) could permit implementation of salvage therapies and improve patient outcomes in acute renal failure (ARF). The utility of single and combined measurements of urinary tubular enzymes in predicting ARF in critically ill patients has not been evaluated using the receiver-operating characteristic (ROC) plot method. METHODS: In this prospective pilot study, 26 consecutive critically ill adult patients admitted to the intensive-care unit were studied. Urine samples were collected twice daily for up to 7 days. ARF was defined as an increase in plasma creatinine of > or = 50% and > or = 0.15 mmol/l. ROC plot analysis was applied to the tubular marker data to derive optimum cut-offs for ARF. RESULTS: Four of the 26 study subjects (15.4%) developed ARF. Indexed to urinary creatinine concentration, gamma glutamyl transpeptidase (gamma GT), alkaline phosphatase (AP), N-acetyl glucosaminidase (NAG), and alpha- and pi-glutathione S-transferase (alpha- and pi GST) but not lactate dehydrogenase (LDH) were higher in the ARF group on admission (P<0.05). gamma GT, and alpha- and pi-GST remained elevated at 24 h. The onset of ARF based on changes in plasma creatinine varied from 12 h to 4 days (median 36 h). ROC plot analysis showed that gamma GT, pi-GST, alpha-GST, AP and NAG had excellent discriminating power for ARF (AUC 0.950, 0.929, 0.893, 0.863 and 0.845, respectively). The discriminating strength of creatinine clearance, while lower, was still significant (AUC 0.796). Positive and negative predictive values for ARF on admission were 67/100% for gamma GT, 67/90% for AP, 60/95% for alpha-GST, and 67/100% for pi-GST indices. Positive and negative predictive values for ARF for creatinine clearance < or = 23 ml/min were 50 and 91%, respectively. Creatinine clearances tended to be lower in ARF than in non-ARF patients on admission (P=0.06) and were significantly lower (P=0.008) after 12 h. Plasma urea and fractional sodium excretion were unhelpful. CONCLUSIONS: Tubular enzymuria on admission to the ICU is useful in predicting ARF. The cheapness and wide availability of automated assays for gamma GT and AP suggests that estimation of these enzymes in random urine samples may be particularly useful for identifying patients at high risk of ARF. PMID- 12584278 TI - Time trends in initiation and dose of dialysis in end-stage renal disease patients in The Netherlands. AB - BACKGROUND: The guidelines published by the NKF-Dialysis Outcomes Quality Initiative (DOQI) in 1997 advocate an earlier start of dialysis in ESRD patients and a higher dialysis dose than usual. We studied the possible influence of the increasing emphasis on adequate dialysis on the management of ESRD patients in The Netherlands in 1993-2000. METHODS: The NECOSAD study on the adequacy of dialysis started in 1993. This prospective multi-centre study included ESRD patients older than 18 years who started HD or PD as the first RRT. We analysed the distribution of age, gender, primary renal disease and co-morbidity, the mean residual renal function and the mean dialysis-Kt/V(urea) at 3 months in 1569 consecutive patients by calendar year of initiation dialysis. RESULTS: Age, gender, primary renal disease and number of co-morbid conditions at the start of dialysis remained stable over time between 1993 and 2000. The mean renal Kt/V(urea) at 3 months increased from 0.5 in 1993 to 0.8 per week in 2000 (P<0.01). An upward trend remained after adjustment for patient characteristics and dialysis centre. The total Kt/V(urea) at 3 months increased from 3.3 in 1993 to 3.7 per week in 2000 in HD (P<0.01) and from 2.0 in 1993 to 2.3 per week in 1999 in PD patients (P<0.01). An upward trend in the dialysis-Kt/V(urea) was found after adjustment for renal Kt/V(urea) (HD: +0.3 per week, P=0.06; PD, +0.2 per week, P<0.05). CONCLUSIONS: These results indicate a tendency towards earlier introduction of RRT and higher doses of dialysis in The Netherlands. Possible effects of this development on mortality, morbidity, quality of life and the balance between costs and benefits need further investigation. PMID- 12584279 TI - Assessment of haemodialysis adequacy by ionic dialysance: intra-patient variability of delivered treatment. AB - INTRODUCTION: Adequate delivered dose of solute removal (as assessed by urea reduction and calculation of Kt/V) is an important determinant of clinical outcome in chronic haemodialysis (HD) patients. The requirement for multiple blood sampling and efforts taken to minimize the effects of rebound on post treatment samples ensure Kt/V is measured only intermittently. On-line conductivity monitoring (using sodium flux as a surrogate for urea) allows the repeated non-invasive measurement of Kt/V on each HD treatment. We have studied the accuracy of this method of measuring Kt/V, and the variability of treatment dose delivered to individual patients. METHODS: We prospectively studied 26 established chronic HD patients over 4 weeks (316 treatments). Patients were dialysed using Hospal Integra dialysis monitors, equipped with Diascan modules to measure Kt/V. Data were downloaded automatically to a central computer server. Urea reduction was measured (once a week) by a two-pool calculation using 30 min post-treatment sampling. RESULTS: Treatment time, Q(B) and modality were fully delivered in all treatments analysed (97% of total). Kt/V measured by ionic dialysance (Kt/V(ID)) correlated highly with that derived from measurement of urea reduction (R(2)=0.92, P<0.0001). Kt/V(ID) underestimated urea-based Kt/V by a mean of only 1.5% (95% CI 0.18-2.9%). Kt/V(ID) varied greatly within individual patients with a mean CV of 0.13+/-0.10 (95% CI 0.05-0.3). If a Kt/V(ID) of 1.0 is considered 'adequate', 55% of the patients had variations that would have potentially altered their status as being adequately or inadequately dialysed, as the range of Kt/V readings cross that point during the study period. CONCLUSION: In conclusion, Kt/V(ID) seems to be an accurate and readily obtained measure of adequacy. Substantial variation in Kt/V implies repeated measures (ideally for all treatments) are necessary to gain a true picture of the mean treatment dose being delivered to patients. PMID- 12584280 TI - Non-dipping is a potent predictor of cardiovascular mortality and is associated with autonomic dysfunction in haemodialysis patients. AB - BACKGROUND: Lack of nocturnal blood pressure (BP) fall (non-dipping) is common among haemodialysis (HD) patients, but much less is known regarding its association with cardiovascular (CV) disease morbidity and mortality. METHODS: Eighty HD patients initially underwent 24 h ambulatory BP monitoring (ABPM), and then they were defined as either 'dippers' (n=24, nocturnal BP fall > or = 10%) or 'non-dippers' (n=56, fall <10%). Coronary angiography was performed in the patients who had signs and/or symptoms of coronary artery disease (CAD). Twenty four hour ambulatory ECG was recorded in 20 dippers and 20 non-dipper HD patients, and in 20 normal subjects. All patients were followed for up to 5.8 years (33.0+/-19.1 months). The outcome events studied were the hospitalisations due to CV diseases and CV death. RESULTS: Compared with dippers, non-dippers initially had a higher incidence of coronary artery stenosis (P<0.05) along with left ventricular asynergy (both Ps<0.01). The circadian rhythm of autonomic function was impaired in non-dippers. The incidences of CV events and CV deaths were 3.5 and 9 times higher in non-dippers than in dippers. The cumulative CV event-free survival and CV survival rates were lower in non-dippers than in dippers (P=0.02 and P=0.005, respectively). Based on Cox analysis, non-dipping was associated positively with CV events and CV mortality [hazard ratio (HR) 2.46, 95% CI 1.02-5.92, P=0.038 and HR 9.62, 95% CI 1.23-75.42, P=0.031, respectively]. Meanwhile, nocturnal systolic BP fall, diurnal systolic BP and diurnal pulse pressure were negatively associated with CV event/death. The clinic BP was not associated with CV event/death. CONCLUSIONS: The non-dipping phenomenon is closely related to a high incidence of CV diseases, a poor long term survival and profound autonomic dysfunction. ABPM is useful in predicting long-term CV prognosis in HD patients. PMID- 12584281 TI - Overuse of acid suppressant drugs in patients with chronic renal failure. AB - BACKGROUND: Patients with chronic renal failure (CRF) have a high prevalence of gastrointestinal symptoms. The use of acid suppressive therapy (AST) in these patients is unexplored. We assessed the use of and indication for AST in CRF patients and compared them with hospitalized patients and patients with another chronic disease. METHODS: A total of 293 patients with CRF were compared with 261 hospitalized patients and 181 patients with another chronic disease. Information about AST, endoscopic investigations and indication for therapy were obtained by medical records and patient interviews. RESULTS: The use of AST was higher in CRF patients (41%, P<0.001) compared with hospitalized patients (13%) and patients with a chronic lung disease (18%), but similar in patients with a rheumatic disease (34%). Inadequate use of AST was high in CRF patients (63%) and hospitalized patients (81%), but lower in patients with another chronic disease (32%, P<0.01). Endoscopic investigations were performed in 41-46% of the patients. More than 90% of AST consisted of proton pump inhibitors (PPIs). CONCLUSION: The majority of CRF patients had an inadequate indication for AST. Endoscopy seemed to be underused in these patients. PPIs were the dominating acid suppressive drug. PMID- 12584282 TI - Correction of anaemia with darbepoetin alfa in patients with chronic kidney disease receiving dialysis. AB - BACKGROUND: Darbepoetin alfa is a new recombinant erythropoietic protein with a 3 fold longer half-life than recombinant human erythropoietin (rHuEpo). The optimal starting dose and frequency of administration of darbepoetin alfa were investigated for treating renal anaemia in dialysis patients. METHODS: Two multicentre, sequential dose-escalation studies examined the i.v. route of administration of darbepoetin alfa in haemodialysis patients (n=75) and the s.c. route in peritoneal dialysis patients (n=47). Patients were randomized to receive darbepoetin alfa at doses ranging from 0.075 to 0.75 microg/kg/week administered as either a once weekly or a three-times weekly injection. Patients achieving the primary endpoint of a > or = 1 g/dl increase in haemoglobin after 4 weeks continued darbepoetin alfa for up to 52 weeks. Safety was assessed by adverse event reports, changes in laboratory values and vital signs, and antibody screening. RESULTS: Darbepoetin alfa produced dose-related increases in haemoglobin over the first 4 weeks of treatment in both studies. Two dose levels (0.45 and 0.75 microg/kg/week) increased the haemoglobin by > or = 1 g/dl in 60 80% of patients, and no difference between once weekly and three-times weekly dosing was apparent. For patients who continued treatment up to 52 weeks, haemoglobin was maintained between 10 and 13 g/dl from mean baseline values of 8.4 and 8.7 g/dl. The adverse event profile was similar to that associated with rHuEpo therapy, and no antibodies to darbepoetin alfa were detected. CONCLUSIONS: Darbepoetin alfa is safe and effective for the treatment of anaemia in dialysis patients. The optimal weekly starting dose is 0.45-0.75 microg/kg and once weekly dosing is possible for both the s.c. and i.v. routes of administration. PMID- 12584283 TI - Sevelamer hydrochloride with or without alphacalcidol or higher dialysate calcium vs calcium carbonate in dialysis patients: an open-label, randomized study. AB - BACKGROUND: Sevelamer hydrochloride was recently proposed as a phosphate binder to prevent hypercalcaemia in place of calcium alkaline salts in dialysis patients. So far, it has been evaluated only in patients receiving calcitriol, without comparison with CaCO(3) alone, although the latter was found to be as effective as the combination of calcitriol and Al(OH)(3) in suppressing parathyroid hormone (PTH) without inducing hypercalcaemia and to have a better lowering effect on serum phosphate. Moreover, this bile salt binder may decrease serum 25-OH vitamin D. Therefore, we compared for 5 months two strategies for controlling moderate hyperparathyroidism: CaCO(3) alone vs sevelamer in conjunction with measures to increase calcium balance. METHODS: Forty-two patients were randomized: 21 continued their treatment with 4.8 g/day CaCO(3) and 21 were switched to sevelamer (initial dose: 2.4 g/day, increased to 4.4 g/day). Each month, when serum-corrected calcium decreased below 2.30 mmol/l, dialysate calcium was increased or alphacalcidol was given at each dialysis session, according to serum PO(4) levels. The following parameters were monitored: serum Ca, PO(4), bicarbonate and protein, weekly; and serum PTH, 25-OH vitamin D and total, LDL and HDL cholesterol monthly. RESULTS: Except for higher serum phosphate at month 1, lower serum bicarbonate at month 2 and lower LDL cholesterol at month 5 in the sevelamer group, no difference was found between the two groups. Compared with baseline levels, PTH increased and 25-OH vitamin D decreased significantly in both groups, these two parameters being inversely correlated. CONCLUSIONS: Given comparable control of plasma calcium, phosphate and 25-OH vitamin D, PTH control is comparable in both strategies. Sevelamer does not induce greater vitamin D depletion than CaCO(3). The transient decrease of serum bicarbonate after discontinuation of CaCO(3) in the sevelamer group suggests a less optimal prevention of acidosis. The sevelamer-induced decrease in LDL cholesterol gives this drug a potential advantage in cardiovascular prevention. PMID- 12584284 TI - Association between residual renal function, inflammation and patient survival in new peritoneal dialysis patients. AB - BACKGROUND: The recent ADEMEX study (Paniagua R, Amato D, Vonesh E et al. J Am Soc Nephrol 2002; 13: 1307-1320) indicates that peritoneal small solute clearance is not as critical for the survival of peritoneal dialysis (PD) patients as thought previously. On the other hand, low residual renal function (RRF), inflammation and an increased peritoneal transport rate (PTR) as evaluated by the peritoneal equilibration test (PET) are reported to be associated with increased mortality in PD patients, but the relationships between these factors and their separate and combined impact on the survival of PD patients are not clear. In this retrospective analysis, we evaluated possible relationships between RRF, inflammation and initial PTR in patients starting PD and the impact of these factors on patient survival. METHODS: A total of 117 patients with initial assessments for RRF, serum C-reactive protein (CRP) and PET at a mean period of 0.4+/-0.2 months (range 0.1-1.0 months) after start of PD were included in this study. Based on RRF (cut-off point, 4 ml/min/1.73 m(2)), serum CRP (cut-off point, 10 mg/l), and the dialysate to plasma creatinine ratio at 4-h of dwell (mean+1 SD), the patients were divided into different groups: low RRF and high RRF group, high CRP and normal CRP group and high PTR and other PTR group, respectively. RESULTS: Of 117 patients, 54 patients (46%) were in low RRF (<4 ml/min/1.73 m(2)) group, 36 patients (31%) were in high serum CRP (> or = 10 mg/l) group and 17 patients (15%) were in high PTR group. Forty-nine patients (42%) had one of these characteristics, 26 patients (22%) had two of these characteristics, two patients (2%) had three, and 40 patients (34%) had none of these characteristics. Patients with low RRF were older and had a higher prevalence of high CRP, lower normalized protein equivalent of total nitrogen appearance (nPNA), lower total Kt/V(urea) and lower total creatinine clearance (CCr) whereas patients with high CRP were older and had a higher proportion of men, lower serum albumin, lower nPNA, lower RRF and lower total CCr. Patients with high PTR had lower serum albumin, higher RRF and higher total CCr compared with patients with other PTR. Upon logistic multiple regression analysis, age and RRF were identified as factors affecting inflammation. Overall patient survival was significantly lower in the patients with low RRF, with high CRP, and in patients with more than two of the following: low RRF, high CRP and high PTR. In contrast, in patients with none of the discriminators low RRF, high CRP and high PTR, the 5-year survival was 100%. A high PTR was associated with decreased survival during the initial year on PD, but not thereafter. Patients who died during the follow-up period had a higher prevalence of high CRP and lower serum albumin, lower RRF, lower Kt/V(urea) and lower total CCr. Upon Cox proportional hazards multivariate analysis, age and RRF were predictors of mortality. CONCLUSIONS: These results indicate that in patients starting PD, low initial RRF is associated with inflammation, and low RRF and inflammation are both associated with high overall mortality. A high PTR was associated with higher mortality, but only during the initial year on PD, whereas Kt/V(urea) did not predict mortality. These results indicate the importance of RRF and inflammation as predictors of mortality in PD patients whereas the predictive power of PTR as such may lose its significance if these two parameters are taken into consideration. PMID- 12584285 TI - Percent-free prostate specific antigen is elevated in men on haemodialysis or peritoneal dialysis treatment. AB - BACKGROUND: Men with chronic renal failure evaluated for transplantation are often tested for prostate specific antigen (PSA) to detect prostate cancer. PSA occurs in several different molecular forms in serum: free PSA (fPSA) and complexed PSA (cPSA), the sum of which corresponds to total PSA (tPSA). In addition to tPSA, percent fPSA to tPSA (%fPSA) is widely used to enhance discrimination of benign disorders from prostate cancer. The low molecular mass of fPSA suggests elimination by renal glomerular filtration and that renal failure may significantly influence %fPSA. We evaluated whether established reference levels for %fPSA are applicable also to patients treated with haemodialysis or continuous ambulatory peritoneal dialysis (CAPD). METHODS: The study included 20 men on intermittent haemodialysis with low-flux membranes and 25 men on CAPD, without known history of prostate cancer. The control group included 3129 men without known prostate cancer. We analysed fPSA and tPSA in serum by dual-label immunofluorometric assays, from which we calculated %fPSA and cPSA. Serum levels of different PSA forms were adjusted for age and presented as geometric means. RESULTS: Percent fPSA was significantly higher in patients on either haemodialysis (39.5%) or CAPD (39.6%) compared with controls (28.1%). Haemodialysis patients, but not CAPD patients, had significantly higher mean levels of fPSA. Levels of tPSA and cPSA for haemodialysis or CAPD patients did not differ significantly compared with controls. CONCLUSIONS: Recommended reference ranges for %fPSA, based on men with normal renal function, do not apply to uraemic men on dialysis. In these men, a high %fPSA should not be considered as a sign of benign disease. This is clinically important in the evaluation of dialysis patients for transplantation, as %fPSA is often used as a tool for detection of prostate cancer. PMID- 12584286 TI - Long-term efficacy of hyperuricaemia treatment in renal transplant patients. AB - BACKGROUND: Although hyperuricaemia and gout are frequently found in renal transplant recipients, little has been published on the efficacy of urate lowering therapy (ULT) in this patient population. We therefore examine the effects of allopurinol and benziodarone therapy in a cohort of renal transplant patients. METHODS: We reviewed files from a cohort of 1328 patients that received renal transplantation. The selection criteria included: functioning allograft, hyperuricaemia for >12 months or gout, ULT lasting at least 1 year and at least two control measurements after the onset of ULT. Patients on azathioprine were treated with benziodarone to avoid azathioprine-allopurinol interactions. RESULTS: Two-hundred and seventy-nine patients fulfilled the criteria for review. They were treated with 289 courses of ULT: 100 with allopurinol (mean dose: 376 mg/day/dl/min of creatinine clearance) and 189 with benziodarone (mean dose: 73 mg/day). The mean follow-up was 38 months. Both drugs were effective for the control of hyperuricaemia, but benziodarone caused greater reductions in serum uric acid levels, especially when used at mean doses of >75 mg/day. Severe side effects were uncommon, in both the allopurinol and benziodarone groups. CONCLUSIONS: Both allopurinol and benziodarone were effective for the control of hyperuricaemia in renal transplantation. Benziodarone at doses >75 mg/day was more effective than allopurinol in reducing serum uric acid levels and also reduced the risk of azathioprine-allopurinol interactions. PMID- 12584287 TI - Effector T cells and macrophages in urine as a hallmark of systemic vasculitis accompanied by crescentic glomerulonephritis. PMID- 12584288 TI - A favourable outcome using oral prednisolone and losartan only in a patient with ANCA-related glomerulonephritis: a discrepancy between histological activity and MPO-ANCA. PMID- 12584289 TI - A well-meant present from a friend. PMID- 12584290 TI - Myoglobinuria: round up more than the usual suspects. PMID- 12584291 TI - Post-traumatic haemodialysis catheter fracture with bacteraemia. PMID- 12584292 TI - Does a reduced 11 beta HSD type 2 activity contribute to sodium retention in the nephrotic syndrome? PMID- 12584294 TI - Evaluating candidates for kidney transplantation: some recommendations still lack convincing clinical evidence. PMID- 12584296 TI - Association of primary hyperparathyroidism with cystinuria in a recurrent renal stone-forming patient. PMID- 12584297 TI - Anaemia among subjects with chronic renal insufficiency. PMID- 12584298 TI - Regular low-intensity aquatic exercise improves cardio-respiratory functional capacity and reduces proteinuria in chronic renal failure patients. PMID- 12584299 TI - Spontaneous renal subcapsular haematoma and acute renal failure complicated by severe pre-eclampsia. PMID- 12584300 TI - Sterile icodextrin-associated peritonitis may induce hypersensitivity and recurrent peritonitis on re-challenge. PMID- 12584303 TI - Viral internal ribosome entry site elements: novel ribosome-RNA complexes and roles in viral pathogenesis. PMID- 12584304 TI - Simian virus 40 small tumor antigen induces deregulation of the actin cytoskeleton and tight junctions in kidney epithelial cells. AB - There is increasing evidence that the transforming DNA tumor virus simian virus 40 (SV40) is associated with human malignancies. SV40 small tumor antigen (small t) interacts with endogenous serine/threonine protein phosphatase 2A (PP2A) and is required for the transforming activity of SV40 in epithelial cells of the lung and kidney. Here, we show that expression of SV40 small t in epithelial MDCK cells induces acute morphological changes and multilayering. Significantly, it also causes severe defects in the biogenesis and barrier properties of tight junctions (TJs) but does not prevent formation of adherens junctions. Small t induced TJ defects are associated with a loss of PP2A from areas of cell-cell contact; altered distribution and reduced amounts of the TJ proteins ZO-1, occludin, and claudin-1; and marked disorganization of the actin cytoskeleton. Small t-mediated F-actin rearrangements encompass increased Rac-induced membrane ruffling and lamellipodia, Cdc42-initiated filopodia, and loss of Rho-dependent stress fibers. Indeed, these F-actin changes coincide with elevated levels of Rac1 and Cdc42 and decreased amounts of RhoA in small t-expressing cells. Notably, these cellular effects of small t are dependent on its interaction with endogenous PP2A. Thus, our findings provide the first evidence that, in polarized epithelial cells, expression of small t alone is sufficient to induce deregulation of Rho GTPases, F-actin, and intercellular adhesion, through interaction with endogenous PP2A. Because defects in the actin cytoskeleton and TJ disruption have been linked to loss of cell polarity and tumor invasiveness, their deregulation by PP2A and small t likely contributes to the role of SV40 in epithelial cell transformation. PMID- 12584305 TI - Human papillomavirus type 31 E5 protein supports cell cycle progression and activates late viral functions upon epithelial differentiation. AB - The function of the E5 protein of human papillomaviruses (HPV) is not well characterized, and controversies exist about its role in the viral life cycle. To determine the function of E5 within the life cycle of HPV type 31 (HPV31) we first constructed HPV31 mutant genomes that contained an altered AUG initiation codon or stop codons in E5. Cell lines were established which harbored transfected wild-type or E5 mutant HPV31 genomes. These cell lines all maintained episomal copies of HPV31 and revealed similar phenotypes with respect to growth rate, early gene expression, and viral copy number in undifferentiated monolayer cultures. Following epithelial differentiation, genome amplification and differentiation-dependent late gene expression were observed in mutant cell lines, but at a rate significantly reduced from that observed in cells containing the wild-type genomes. Organotypic raft cultures indicated that E5 does not effect the expression of differentiation markers but does reduce expression of late viral proteins. Western analysis and immunofluorescence staining for cyclins during epithelial differentiation revealed a decreased expression of cyclin A and B in E5 mutant cells compared to HPV wild-type cells. Using a replating assay, a significant reduction in colony-forming ability was detected in the absence of E5 expression when cells containing wild-type or E5 mutant HPV genomes were allowed to proliferate following 24 h in suspension-induced differentiation. This suggests that HPV E5 modifies the differentiation-induced cell cycle exit and supports the ability of HPV31-positive keratinocytes to retain proliferative competence. In these studies, E5 was found to have little effect on the levels of the epidermal growth factor receptor (EGFR) or on its phosphorylation status. This indicates that EGFR is not a target of E5 action. Our results propose a role for high risk HPV E5 in modulation of late viral functions through activation of proliferative capacity in differentiated cells. We suspect that the primary target of E5 is a membrane protein or receptor that then acts to alter the levels or activities of cell cycle regulators. PMID- 12584306 TI - Quantitative role of the human papillomavirus type 16 E5 gene during the productive stage of the viral life cycle. AB - Human papillomaviruses (HPVs) are small circular DNA viruses that cause warts. Infection with high-risk anogenital HPVs, such as HPV type 16 (HPV16), is associated with human cancers, specifically cervical cancer. The life cycle of HPVs is intimately tied to the differentiation status of the host epithelium and has two distinct stages: the nonproductive stage and the productive stage. In the nonproductive stage, which arises in the poorly differentiated basal epithelial compartment of a wart, the virus maintains itself as a low-copy-number nuclear plasmid. In the productive stage, which arises as the host cell undergoes terminal differentiation, viral DNA is amplified; the capsid genes, L1 and L2, are expressed; and progeny virions are produced. This stage of the viral life cycle relies on the ability of the virus to reprogram the differentiated cells to support DNA synthesis. Papillomaviruses encode multiple oncoproteins, E5, E6, and E7. In the present study, we analyze the role of one of these viral oncogenes, E5, in the viral life cycle. To assess the role of E5 in the HPV16 life cycle, we introduced wild-type (WT) or E5 mutant HPV16 genomes into NIKS, a keratinocyte cell line that supports the papillomavirus life cycle. By culturing these cells under conditions that allow them to remain undifferentiated, a state similar to that of basal epithelial cells, we determined that E5 does not play an essential role in the nonproductive stage of the HPV16 life cycle. To determine if E5 plays a role in the productive stage of the viral life cycle, we cultured keratinocyte populations in organotypic raft cultures, which promote the differentiation and stratification of epithelial cells. We found that cells harboring E5 mutant genomes displayed a quantitative reduction in the percentage of suprabasal cells undergoing DNA synthesis, compared to cells containing WT HPV16 DNA. This reduction in DNA synthesis, however, did not prevent amplification of viral DNA in the differentiated cellular compartment. Likewise, late viral gene expression and the perturbation of normal keratinocyte differentiation were retained in cells harboring E5 mutant genomes. These data demonstrate that E5 plays a subtle role during the productive stage of the HPV16 life cycle. PMID- 12584307 TI - Leader proteinase of beet yellows virus functions in long-distance transport. AB - The 66-kDa leader proteinase (L-Pro) of the Beet yellows virus (BYV) possesses a nonconserved N-terminal domain and a conserved, papain-like C-terminal domain. Previous work revealed that the N-terminal domain functions in RNA amplification, whereas the C-terminal domain is required for autoproteolysis. Alanine-scanning mutagenesis was applied to complete the functional analysis of L-Pro throughout the virus life cycle. This analysis indicated that the C-terminal domain of L Pro, in addition to being required for proteolysis, also functions in RNA amplification and that these two functions are genetically separable. Examination of the role of L-Pro in BYV cell-to-cell movement revealed that none of the 20 examined replication-competent mutants was movement defective. In contrast, six of the L-Pro mutations affected the long-distance transport of BYV to various degrees, whereas three mutations completely abolished the transport. Because these mutations were located throughout the protein molecule, both domains of L Pro function in virus transport. We conclude that in addition to previously identified functions of L-Pro, it also serves as the BYV long-distance transport factor. PMID- 12584308 TI - Role of virus receptor Hyal2 in oncogenic transformation of rodent fibroblasts by sheep betaretrovirus env proteins. AB - The ovine betaretroviruses jaagsiekte sheep retrovirus (JSRV) and enzootic nasal tumor virus (ENTV) cause contagious cancers in the lungs and upper airways of sheep and goats. Oncogenic transformation assays using mouse and rat fibroblasts have localized the transforming activity to the Env proteins encoded by these viruses, which require the putative lung and breast cancer tumor suppressor hyaluronidase 2 (Hyal2) to promote virus entry into cells. These results suggested the hypothesis that the JSRV and ENTV Env proteins cause cancer by inhibiting the tumor suppressor activity of Hyal2. Consistent with this hypothesis, we show that human Hyal2 and other Hyal2 orthologs that can promote virus entry, including rat Hyal2, can suppress transformation by the Env proteins of JSRV and ENTV. Furthermore, we provide direct evidence for binding of the surface (SU) region of JSRV Env to human and rat Hyal2. However, mouse Hyal2 did not mediate entry of virions bearing JSRV or ENTV Env proteins, bound JSRV SU poorly if at all, and did not suppress transformation by the JSRV or ENTV Env proteins, indicating that mouse Hyal2 plays no role in transformation of mouse fibroblasts and that the Env proteins can transform at least some cells by a Hyal2-independent mechanism. Expression of human Hyal2 in mouse cells expressing JSRV Env caused a marked reduction in Env protein levels, indicating that human Hyal2 suppresses Env-mediated transformation in mouse cells by increasing Env degradation rather than by exerting a more general Env-independent tumor suppressor activity. PMID- 12584309 TI - Neutralization of human immunodeficiency virus type 1 by sCD4-17b, a single-chain chimeric protein, based on sequential interaction of gp120 with CD4 and coreceptor. AB - We designed a novel single-chain chimeric protein, designated sCD4-17b, for neutralization of human immunodeficiency virus type 1 (HIV-1). The recombinant protein contains domains 1 and 2 of soluble CD4 (sCD4), connected via a flexible polypeptide linker to a single-chain variable region construct of 17b, a human monoclonal antibody that targets a conserved CD4-induced epitope on gp120 overlapping the coreceptor binding region. We hypothesized that the sCD4 moiety would bind gp120 and expose the 17b epitope; the 17b moiety would then bind, thereby blocking coreceptor interaction and neutralizing infection. The sCD4-17b protein, expressed by a recombinant vaccinia virus, potently neutralized a prototypic R5 clade B primary isolate, with a 50% inhibitory concentration of 3.2 nM (0.16 microg/ml) and >95% neutralization at 32 nM (1.6 microg/ml). The individual components (sCD4 and 17b, singly or in combination) had minimal effects at these concentrations, demonstrating that the activity of sCD4-17b reflected the ability of a single chimeric molecule to bind gp120 simultaneously via two independent moieties. sCD4-17b was highly potent compared to the previously characterized broadly cross-reactive neutralizing monoclonal antibodies IgGb12, 2G12, and 2F5. Multiple primary isolates were neutralized, including two previously described as antibody resistant. Neutralization occurred for both R5 and X4 strains and was not restricted to clade B. However, several primary isolates were insensitive over the concentration range tested, despite the known presence of binding sites for both CD4 and 17b. sCD4-17b has potential utility for passive immunization against HIV-1 in several contexts, including maternal transmission, postexposure prophylaxis, and sexual transmission (topical microbicide). PMID- 12584310 TI - Endoproteolytic processing of the lymphocytic choriomeningitis virus glycoprotein by the subtilase SKI-1/S1P. AB - The envelope glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) is posttranslationally cleaved into two subunits. We show here that this endoproteolytic processing is not required for transport to the cell surface but is essential for LCMV GP to mediate infectivity of pseudotyped retroviral vectors. By systematic mutational analysis of the LCMV GP cleavage site, we determined that the consensus motif R-(R/K/H)-L-(A/L/S/T/F)(265) is essential for the endoproteolytic processing. In agreement with the identified consensus motif, we show that the cellular subtilase SKI-1/S1P cleaves LCMV GP. PMID- 12584311 TI - Presence of replicating virus in recombinant hepadnavirus stocks results from recombination and can be eliminated by the use of a packaging cell line. AB - Mutant hepatitis B viruses are useful tools to study the viral life cycle and viral pathogenesis. Furthermore, recombinant hepatitis B viruses are candidate vectors for liver-directed gene therapy. Because wild-type viruses present in recombinant or mutant virus stocks may falsify experimental results and are detrimental for a viral vector, we investigated whether and to what extent wild type virus is present in recombinant virus stocks and where it originates from. We took advantage of the duck model of hepatitis B virus infection which allows very sensitive detection of replication-competent viruses by infection of primary duck hepatocytes or of ducklings in vivo. Recombinant hepatitis B virus stocks contained significant amounts of wild-type viruses, which were most probably generated by homologous recombination between plasmids containing homologous viral sequences. In addition, replication-competent viral genomes were reconstituted from plasmids which contained replication-deficient but redundant viral sequences. Using a stable cell line for packaging of deficient viral genomes, no wild-type virus was detected, neither by infection of primary hepatocytes nor in vivo. PMID- 12584312 TI - In vitro and in vivo characterization of a murine cytomegalovirus with a mutation at open reading frame m166. AB - We have recently generated a pool of murine cytomegalovirus (MCMV) mutants by using a Tn3-based transposon mutagenesis approach. In this study, one of the mutants, Rvm166, which contained the transposon sequence at open reading frame m166, was characterized both in tissue culture and in immunocompetent BALB/c mice and immunodeficient SCID mice. The viral mutant replicated as well as the wild type Smith strain in vitro in NIH 3T3 cells, whereas the transposon insertion precluded the expression of >65% of the m166 open reading frame. Compared to the wild-type strain and a rescued virus that restored the m166 region, the viral mutant was significantly attenuated in growth in both BALB/c and SCID mice that were intraperitoneally infected with the viruses. At 21 days postinfection, the titers of the viral mutant in the salivary glands, lungs, spleens, livers, and kidneys of the infected SCID mice were lower than the titers of the Smith strain and the rescued virus by about 30000-, 10000-, 1000-, 300-, and 800-fold, respectively. Moreover, the virulence of the mutant virus appears to be severely attenuated because no death was found in SCID mice infected with the viral mutant up to 90 days postinfection, whereas all of the animals infected with the wild type and rescued viruses died at 27 days postinfection. Our results suggest that m166 probably encodes a virulence factor and is required for MCMV virulence in killing SCID mice and for optimal viral growth in vivo. PMID- 12584313 TI - A single amino acid substitution in herpes simplex virus type 1 VP16 inhibits binding to the virion host shutoff protein and is incompatible with virus growth. AB - In addition to its well-established role in the activation of herpes simplex virus immediate-early gene transcription, VP16 interacts with and downregulates the function of the virion host shutoff protein (vhs), thereby attenuating vhs mediated destruction of viral mRNAs and translational arrest at late times of infection. We have carried out two-hybrid analysis in vivo and protein-protein interaction assays in vitro to identify determinants in VP16 necessary for interaction with vhs. The minimal amino-terminal subfragment of VP16 capable of binding to vhs encompassed residues 1 to 345. Alteration of a single leucine at position 344 to alanine (L344A) in the context of the amino-terminal fragment of VP16 containing residues 1 to 404 was sufficient to abolish interaction with vhs in vitro and in vivo. Leu344 could be replaced with hydrophobic amino acids (Ile, Phe, Met, or Val) but not by Asn, Lys, or Pro, indicating that hydrophobicity is an important property of binding to vhs. VP16 harboring a loss-of-function mutation at L344 was not compromised in its ability to interact with host cell factor (HCF-1) or to activate transcription of viral immediate-early genes in transient-transfection assays. Virus complementation assays using the VP16-null virus 8MA and the VP16/vhs double-mutant virus 8MAdeltaSma showed that VP16(L344A) was able to complement the growth of 8MAdeltaSma but not 8MA. Thus, a single point mutation in VP16 uncouples binding to vhs from other functions of VP16 required for virus growth and indicates that direct physical association between VP16 and vhs is necessary to sustain a productive infection. PMID- 12584314 TI - Transcription factor YY1 binds to the murine beta interferon promoter and regulates its transcriptional capacity with a dual activator/repressor role. AB - The induction of the beta interferon (IFN-beta) gene constitutes one of the first responses of the cell to virus infection. Its regulation is achieved through an intricate combination of virus-induced binding of transcription factors and local chromatin remodeling. In this work, we demonstrate that transcription factor YY1, known to interact with histone deacetylases (HDAC) and histone acetyltransferases, has a dual activator/repressor role during the regulation of the IFN-beta promoter activity. We show that YY1 specifically binds in vitro and in vivo to the murine IFN-beta promoter at positions -90 and -122. Overexpression of YY1 strongly repressed the transcriptional capacity of a stably integrated IFN beta promoter fused to a chloramphenicol acetyltransferase reporter gene as well as the endogenous IFN activity of murine L929 cells via an HDAC activity. Stably integrated IFN-beta promoters mutated at the -90 site were no longer repressed by YY1, could no longer be activated by trichostatin A, displayed a retarded postinduction turn off, and a reduced virus-induced activity. Introduction of a mutation at the -122 site did not affect YY1-induced repression, but promoters with this mutation displayed a reduced virus-induced activity. Stably integrated full-length promoters (from position -330 to +20) mutated at both YY1-binding sites displayed extremely reduced promoter activities. We conclude that YY1 has a dual activator/repressor role on IFN-beta promoter activity depending on its binding site and time after infection. PMID- 12584315 TI - Human parvovirus B19 nonstructural protein (NS1) induces cell cycle arrest at G(1) phase. AB - Human parvovirus B19 infects predominantly erythroid precursor cells, leading to inhibition of erythropoiesis. This erythroid cell damage is mediated by the viral nonstructural protein 1 (NS1) through an apoptotic mechanism. We previously demonstrated that B19 virus infection induces G(2) arrest in erythroid UT7/Epo-S1 cells; however, the role of NS1 in regulating cell cycle arrest is unknown. In this report, by using paclitaxel, a mitotic inhibitor, we show that B19 virus infection induces not only G(2) arrest but also G(1) arrest. Interestingly, UV irradiated B19 virus, which has inactivated the expression of NS1, still harbors the ability to induce G(2) arrest but not G(1) arrest. Furthermore, treatment with caffeine, a G(2) checkpoint inhibitor, abrogated the B19 virus-induced G(2) arrest despite expression of NS1. These results suggest that the B19 virus induced G(2) arrest is not mediated by NS1 expression. We also found that NS1 transfected UT7/Epo-S1 and 293T cells induced cell cycle arrest at the G(1) phase. These results indicate that NS1 expression plays a critical role in G(1) arrest induced by B19 virus. Furthermore, NS1 expression significantly increased p21/WAF1 expression, a cyclin-dependent kinase inhibitor that induces G(1) arrest. Thus, G(1) arrest mediated by NS1 may be a prerequisite for the apoptotic damage of erythroid progenitor cells upon B19 virus infection. PMID- 12584316 TI - Nucleocapsid-independent specific viral RNA packaging via viral envelope protein and viral RNA signal. AB - For any of the enveloped RNA viruses studied to date, recognition of a specific RNA packaging signal by the virus's nucleocapsid (N) protein is the first step described in the process of viral RNA packaging. In the murine coronavirus a selective interaction between the viral transmembrane envelope protein M and the viral ribonucleoprotein complex, composed of N protein and viral RNA containing a short cis-acting RNA element, the packaging signal, determines the selective RNA packaging into virus particles. In this report we show that expressed coronavirus envelope protein M specifically interacted with coexpressed noncoronavirus RNA transcripts containing the short viral packaging signal in the absence of coronavirus N protein. Furthermore, this M protein-packaging signal interaction led to specific packaging of the packaging signal-containing RNA transcripts into coronavirus-like particles in the absence of N protein. These findings not only highlight a novel RNA packaging mechanism for an enveloped virus, where the specific RNA packaging can occur without the core or N protein, but also point to a new, biologically important general model of precise and selective interaction between transmembrane proteins and specific RNA elements. PMID- 12584317 TI - Persistent replication of hepatitis C virus replicons expressing the beta lactamase reporter in subpopulations of highly permissive Huh7 cells. AB - Progress toward development of better therapies for the treatment of hepatitis C virus (HCV) infection has been hampered by poor understanding of HCV biology and the lack of biological assays suitable for drug screening. Here we describe a powerful HCV replication system that employs HCV replicons expressing the beta lactamase reporter (bla replicons) and subpopulations of Huh7 cells that are more permissive (or "enhanced") to HCV replication than naive Huh7 cells. Enhanced cells represent a small fraction of permissive cells present among naive Huh7 cells that is enriched during selection with replicons expressing the neomycin phosphotransferase gene (neo replicons). The level of permissiveness of cell lines harboring neo replicons can vary greatly, and the enhanced phenotype is usually revealed upon removal of the neo replicon with inhibitors of HCV replication. Replicon removal is responsible for increased permissiveness, since this effect could be reproduced either with alpha interferon or with an HCV NS5B inhibitor. Moreover, adaptive mutations present in the replicon genome used during selection do not influence the permissiveness of the resulting enhanced cell population, suggesting that the mechanisms governing the permissiveness of enhanced cells are independent from viral adaptation. Because the beta-lactamase reporter allows simultaneous quantitation of replicon-harboring cells and reporter activity, it was possible to investigate the relationship between genome replication activity and the frequency with which transfected genomes can establish persistent replication. Our study demonstrates that differences in the replication potential of the viral genome are manifested primarily in the frequency with which persistent replication is established but modestly affect the number of replicons observed per replicon-harboring cell. Replicon copy number was found to vary over a narrow range that may be defined by a minimal number required for persistent maintenance and a maximum that is limited by the availability of essential host factors. PMID- 12584318 TI - N-linked glycosylation and sequence changes in a critical negative control region of the ASCT1 and ASCT2 neutral amino acid transporters determine their retroviral receptor functions. AB - A widely dispersed interference group of retroviruses that includes the feline endogenous virus (RD114), baboon endogenous virus (BaEV), human endogenous virus type W (HERV-W), and type D primate retroviruses uses the human Na(+)-dependent neutral amino acid transporter type 2 (hASCT2; gene name, SLC1A5) as a common cell surface receptor. Although hamster cells are fully resistant to these viruses and murine cells are susceptible only to BaEV and HERV-W pseudotype viruses, these rodent cells both become highly susceptible to all of the viruses after treatment with tunicamycin, an inhibitor of protein N-linked glycosylation. A partial explanation for these results was recently provided by findings that the orthologous murine transporter mASCT2 is inactive as a viral receptor, that a related (ca. 55% identity) murine paralog (mASCT1; gene name, SLC1A4) mediates infections specifically of BaEV and HERV-W, and that N-deglycosylation of mASCT1 activates it as a receptor for all viruses of this interference group. Because the only two N-linked oligosaccharides in mASCT1 occur in the carboxyl-terminal region of extracellular loop 2 (ECL2), it was inferred that this region contributes in an inhibitory manner to infections by RD114 and type D primate viruses. To directly and more thoroughly investigate the receptor active sites, we constructed and analyzed a series of hASCT2/mASCT2 chimeras and site-directed mutants. Our results suggest that a hypervariable sequence of 21 amino acids in the carboxyl-terminal portion of ECL2 plays a critical role in determining the receptor properties of ASCT2 proteins for all viruses in this interference group. In addition, we analyzed the tunicamycin-dependent viral susceptibility of hamster cells. In contrast to mASCT1, which contains two N-linked oligosaccharides that partially restrict viral infections, hamster ASCT1 contains an additional N-linked oligosaccharide clustered close to the others in the carboxyl-terminal region of ECL2. Removal of this N-linked oligosaccharide by mutagenesis enabled hamster ASCT1 to function as a receptor for all viruses of this interference group. These results strongly suggest that combinations of amino acid sequence changes and N-linked oligosaccharides in a critical carboxyl terminal region of ECL2 control retroviral utilization of both the ASCT1 and ASCT2 receptors. PMID- 12584319 TI - Exonuclease-deficient polymerase mutant of herpes simplex virus type 1 induces altered spectra of mutations. AB - The effect of exonuclease activity of the herpes simplex virus DNA polymerase (Pol) on DNA replication fidelity was examined by using the supF mutagenesis assay. The recombinants with exonuclease-deficient Pol, containing an integrated supF gene in the thymidine kinase locus (tk), exhibited supF mutation frequencies ranging from 0.14 to 5.6%, consistent with the tk mutation frequencies reported previously (Y. T. Hwang, B.-Y. Liu, D. M. Coen, and C. B. C. Hwang, J. Virol. 71:7791-7798, 1997). The increased mutation frequencies were 10- to 500-fold higher than those observed for wild-type Pol recombinants. The increased mutation frequencies also were significantly higher than those of supF mutant replicated by exonuclease-deficient Pols in the plasmid-borne assay. Furthermore, characterization of supF mutants demonstrated that recombinants with a defective exonuclease induced types and distributions of supF mutations different from those induced by wild-type Pol recombinants. The types of supF mutations induced by exonuclease-deficient recombinants differed between the plasmid- and genome based assays. The spectra of supF mutations also differed between the two assays. In addition, exonuclease-defective viruses also induced different spectra of supF and tk mutations. Therefore, both the assay methods and the target genes used for mutagenesis studies can affect the repication fidelity of herpes simplex virus type 1 Pol with defective exonuclease activity. PMID- 12584320 TI - Expansion of human T-cell leukemia virus type 1 (HTLV-1) reservoir in orally infected rats: inverse correlation with HTLV-1-specific cellular immune response. AB - Adult T-cell leukemia (ATL) occurs in a small population of human T-cell leukemia virus type 1 (HTLV-1)-infected individuals. Although the critical risk factor for ATL development is not clear, it has been noted that ATL is incidentally associated with mother-to-child infection, elevated proviral loads, and weakness in HTLV-1-specific T-cell immune responses. In the present study, using a rat system, we investigated the relationships among the following conditions: primary HTLV-1 infection, a persistent HTLV-1 load, and host HTLV-1-specific immunity. We found that the persistent HTLV-1 load in orally infected rats was significantly greater than that in intraperitoneally infected rats. Even after inoculation with only 50 infected cells, a persistent viral load built up to considerable levels in some orally infected rats but not in intraperitoneally infected rats. In contrast, HTLV-1-specific cellular immune responses were markedly impaired in orally infected rats. As a result, a persistent viral load was inversely correlated with levels of virus-specific T-cell responses in these rats. Otherwise very weak HTLV-1-specific cellular immune responses in orally infected rats were markedly augmented after subcutaneous reimmunization with infected syngeneic rat cells. These findings suggest that HTLV-1-specific immune unresponsiveness associated with oral HTLV-1 infection may be a potential risk factor for development of ATL, allowing expansion of the infected cell reservoir in vivo, but could be overcome with immunological strategies. PMID- 12584321 TI - A new hybrid system capable of efficient lentiviral vector production and stable gene transfer mediated by a single helper-dependent adenoviral vector. AB - To achieve efficient and sustained gene expression, we developed a new lentivirus/adenovirus hybrid vector (LA vector) that encodes sequences required for production of a human immunodeficiency virus-based lentiviral vector (i.e., a lentiviral vector, a gag/pol/rev expression cassette, a tetracycline-inducible envelope cassette, and the tetracycline-inducible transcriptional activator cassette) in a single helper-dependent adenovirus vector backbone. Via either transfection or infection, human cell lines transduced with the LA vector produced a lentiviral vector in a doxycycline-dependent manner at titers up to 10(5) to 10(6) green fluorescent protein transducing units per ml, which are comparable to the titers obtained by conventional multiple plasmid transfection methods. Efficient spread and persistent expression of the transgene were observed in cells maintained in long-term culture that had been infected with the LA vector. Furthermore, when cocultured with adherent cells infected with the LA vector, the human T-cell leukemia cell line was successfully transduced with a marker gene. This LA vector possesses the advantages of efficient gene transfer from an adenoviral vector and stable integration from a lentiviral vector; therefore, it might have potential for a variety of gene therapy applications. PMID- 12584322 TI - Multitasking in replication is common among geminiviruses. AB - Geminiviruses package single-stranded circular DNA and replicate via double stranded DNA intermediates. During the past decade, increasing evidence has led to the general acceptance that their replication follows a rolling-circle replication mechanism like bacteriophages with single-stranded DNA. In a recent study, we showed that this is also true for Abutilon mosaic geminivirus (AbMV), but that this particular virus may also use a recombination-dependent replication (RDR) route in analogy to T4 phages. Because AbMV is a special case, since it has been propagated on ornamental plants for more than a hundred years, it was interesting to determine whether RDR is common among other geminiviruses. We analyzed geminiviruses from different genera and geographic origins by using BND cellulose chromatography in combination with an improved high resolution two dimensional gel electrophoresis, and we conclude that multitasking in replication is widespread, at least for African cassava mosaic, Beet curly top, Tomato golden mosaic, and Tomato yellow leaf curl virus. PMID- 12584323 TI - Furin processing and proteolytic activation of Semliki Forest virus. AB - The alphavirus Semliki Forest virus (SFV) infects cells via a low-pH-dependent membrane fusion reaction mediated by the E1 envelope protein. Fusion is regulated by the interaction of E1 with the receptor-binding protein E2. E2 is synthesized as a precursor termed "p62," which forms a stable heterodimer with E1 and is processed late in the secretory pathway by a cellular furin-like protease. Once processing to E2 occurs, the E1/E2 heterodimer is destabilized so that it is more readily dissociated by exposure to low pH, allowing fusion and infection. We have used FD11 cells, a furin-deficient CHO cell line, to characterize the processing of p62 and its role in the control of virus fusion and infection. p62 was not cleaved in FD11 cells and cleavage was restored in FD11 cell transfectants expressing human furin. Studies of unprocessed virus produced in FD11 cells (wt/p62) demonstrated that the p62 protein was efficiently cleaved by purified furin in vitro, without requiring prior exposure to low pH. wt/p62 virus particles were also processed during their endocytic uptake in furin-containing cells, resulting in more efficient virus infection. wt/p62 virus was compared with mutant L, in which p62 cleavage was blocked by mutation of the furin recognition motif. wt/p62 and mutant L had similar fusion properties, requiring a much lower pH than control virus to trigger fusion and fusogenic E1 conformational changes. However, the in vivo infectivity of mutant L was more strongly inhibited than that of wt/p62, due to additional effects of the mutation on virus-cell binding. PMID- 12584324 TI - Mutation of host DnaJ homolog inhibits brome mosaic virus negative-strand RNA synthesis. AB - The replication of positive-strand RNA viruses involves not only viral proteins but also multiple cellular proteins and intracellular membranes. In both plant cells and the yeast Saccharomyces cerevisiae, brome mosaic virus (BMV), a member of the alphavirus-like superfamily, replicates its RNA in endoplasmic reticulum (ER)-associated complexes containing viral 1a and 2a proteins. Prior to negative strand RNA synthesis, 1a localizes to ER membranes and recruits both positive strand BMV RNA templates and the polymerase-like 2a protein to ER membranes. Here, we show that BMV RNA replication in S. cerevisiae is markedly inhibited by a mutation in the host YDJ1 gene, which encodes a chaperone Ydj1p related to Escherichia coli DnaJ. In the ydj1 mutant, negative-strand RNA accumulation was inhibited even though 1a protein associated with membranes and the positive strand RNA3 replication template and 2a protein were recruited to membranes as in wild-type cells. In addition, we found that in ydj1 mutant cells but not wild type cells, a fraction of 2a protein accumulated in a membrane-free but insoluble, rapidly sedimenting form. These and other results show that Ydj1p is involved in forming BMV replication complexes active in negative-strand RNA synthesis and suggest that a chaperone system involving Ydj1p participates in 2a protein folding or assembly into the active replication complex. PMID- 12584325 TI - Inhibition of adaptive Vgamma2Vdelta2+ T-cell responses during active mycobacterial coinfection of simian immunodeficiency virus SIVmac-infected monkeys. AB - Adaptive immune responses of gammadelta T cells during active mycobacterial coinfection of human immunodeficiency virus-infected humans have not been studied. Macaques infected with the simian immunodeficiency virus (SIV) SIVmac were employed to determine the extent to which a coincident AIDS virus infection might compromise immune responses of mycobacterium-specific Vgamma2Vdelta2(+) T cells during active mycobacterial infection. Control SIVmac-negative macaques developed primary and recall expansions of phosphoantigen-specific Vgamma2Vdelta2(+) T cells after Mycobacterium bovis BCG infection and BCG reinfection, respectively. In contrast, SIVmac-infected macaques did not exhibit sound primary and recall expansions of Vgamma2Vdelta2(+) T cells in the blood and pulmonary alveoli following BCG infection and reinfection. The absence of adaptive Vgamma2Vdelta2(+) T-cell responses was associated with profound CD4(+) T cell deficiency and subsequent development of SIVmac-related tuberculosis-like disease in the coinfected monkeys. Consistently, Vgamma2Vdelta2(+) T cells from coinfected monkeys displayed a reduced capacity to expand in vitro following stimulation with phosphoantigen. The reduced ability of Vgamma2Vdelta2(+) peripheral blood lymphocytes (PBL) to expand could be restored to some extent by coculture of these cells with CD4(+) T cells purified from PBL of SIV-negative monkeys. Furthermore, naive monkeys inoculated simultaneously with SIVmac and BCG were unable to sustain expansion of Vgamma2Vdelta2(+) T cells at the time that the coinfected monkeys developed lymphoid depletion and a fatal tuberculosis-like disease. Nevertheless, no deletion in Vdelta2 T-cell receptor repertoire was identified in SIVmac-BCG-coinfected macaques, implicating an SIVmac-induced down regulation rather than a clonal exhaustion of these cells. Thus, an SIVmac induced compromise of the adaptive Vgamma2Vdelta2(+) T-cell responses may contribute to the immunopathogenesis of the SIV-related tuberculosis-like disease in macaques. PMID- 12584326 TI - Viral and cellular determinants of hepatitis C virus RNA replication in cell culture. AB - Studies on the replication of hepatitis C virus (HCV) have been facilitated by the development of selectable subgenomic replicons replicating in the human hepatoma cell line Huh-7 at a surprisingly high level. Analysis of the replicon population in selected cells revealed the occurrence of cell culture-adaptive mutations that enhance RNA replication substantially. To gain a better understanding of HCV cell culture adaptation, we characterized conserved mutations identified by sequence analysis of 26 independent replicon cell clones for their effect on RNA replication. Mutations enhancing replication were found in nearly every nonstructural (NS) protein, and they could be subdivided into at least two groups by their effect on replication efficiency and cooperativity: (i). mutations in NS3 with a low impact on replication but that enhanced replication cooperatively when combined with highly adaptive mutations and (ii). mutations in NS4B, -5A, and -5B, causing a strong increase in replication but being incompatible with each other. In addition to adaptive mutations, we found that the host cell plays an equally important role for efficient RNA replication. We tested several passages of the same Huh-7 cell line and found up to 100-fold differences in their ability to support replicon amplification. These differences were not due to variations in internal ribosome entry site-dependent translation or RNA degradation. In a search for cellular factor(s) that might be responsible for the different levels of permissiveness of Huh-7 cells, we found that replication efficiency decreased with increasing amounts of transfected replicon RNA, indicating that viral RNA or proteins are cytopathic or that host cell factors in Huh-7 cells limit RNA amplification. In summary, these data show that the efficiency of HCV replication in cell culture is determined both by adaptation of the viral sequence and by the host cell itself. PMID- 12584327 TI - Dimerization and template switching in the 5' untranslated region between various subtypes of human immunodeficiency virus type 1. AB - The human immunodeficiency virus type 1 (HIV-1) particle contains two identical RNA strands, each corresponding to the entire genome. The 5' untranslated region (UTR) of each RNA strand contains extensive secondary and tertiary structures that are instrumental in different steps of the viral replication cycle. We have characterized the 5' UTRs of nine different HIV-1 isolates representing subtypes A through G and, by comparing their homodimerization and heterodimerization potentials, found that complementarity between the palindromic sequences in the dimerization initiation site (DIS) hairpins is necessary and sufficient for in vitro dimerization of two subtype RNAs. The 5' UTR sequences were used to design donor and acceptor templates for a coupled in vitro dimerization-reverse transcription assay. We showed that template switching during reverse transcription is increased with a matching DIS palindrome and further stimulated proportional to the level of homology between the templates. The presence of the HIV-1 nucleocapsid protein NCp7 increased the template-switching efficiency for matching DIS palindromes twofold, whereas the recombination efficiency was increased sevenfold with a nonmatching palindrome. Since NCp7 did not effect the dimerization of nonmatching palindromes, we concluded that the protein most likely stimulates the strand transfer reaction. An analysis of the distribution of template-switching events revealed that it occurs throughout the 5' UTR. Together, these results demonstrate that the template switching of HIV-1 reverse transcriptase occurs frequently in vitro and that this process is facilitated mainly by template proximity and the level of homology. PMID- 12584328 TI - An umbraviral protein, involved in long-distance RNA movement, binds viral RNA and forms unique, protective ribonucleoprotein complexes. AB - Umbraviruses are different from most other viruses in that they do not encode a conventional capsid protein (CP); therefore, no recognizable virus particles are formed in infected plants. Their lack of a CP is compensated for by the ORF3 protein, which fulfils functions that are provided by the CPs of other viruses, such as protection and long-distance movement of viral RNA. When the Groundnut rosette virus (GRV) ORF3 protein was expressed from Tobacco mosaic virus (TMV) in place of the TMV CP [TMV(ORF3)], in infected cells it interacted with the TMV RNA to form filamentous ribonucleoprotein (RNP) particles that had elements of helical structure but were not as uniform as classical virions. These RNP particles were observed in amorphous inclusions in the cytoplasm, where they were embedded within an electron-dense matrix material. The inclusions were detected in all types of cells and were abundant in phloem-associated cells, in particular companion cells and immature sieve elements. RNP-containing complexes similar in appearance to the inclusions were isolated from plants infected with TMV(ORF3) or with GRV itself. In vitro, the ORF3 protein formed oligomers and bound RNA in a manner consistent with its role in the formation of RNP complexes. It is suggested that the cytoplasmic RNP complexes formed by the ORF3 protein serve to protect viral RNA and may be the form in which it moves through the phloem. Thus, the RNP particles detected here represent a novel structure which may be used by umbraviruses as an alternative to classical virions. PMID- 12584329 TI - Nef-mediated disruption of HLA-A2 transport to the cell surface in T cells. AB - Human immunodeficiency virus type 1 (HIV-1) Nef is a key pathogenic factor necessary for the development of AIDS. One important function of Nef is to reduce cell surface levels of major histocompatibility complex class I (MHC-I) molecules, thereby protecting HIV-infected cells from recognition by cytotoxic T lymphocytes. The mechanism of MHC-I downmodulation by Nef has not been clearly elucidated, and its reported effect on MHC-I steady-state levels ranges widely, from 2-fold in HeLa cells to 200-fold in HIV-infected primary T cells. Here, we directly compared downmodulation of HLA-A2 in HIV-infected HeLa cells to that in T cells. We found that similar amounts of Nef protein resulted in a much more dramatic downmodulation of HLA-A2 in T cells than in HeLa cells. A comparison of Nef's effects on HLA-A2 endocytosis, recycling, and transport rates indicated that the most prominent effect of Nef on HLA-A2 in T cells was to inhibit transport to the cell surface. The phosphatidylinositol 3-kinase inhibitor, LY294002, previously reported to inhibit Nef-mediated MHC-I downmodulation in astrocytic cells, did not directly affect Nef's ability to block transport of MHC I to the cell surface in T cells. PMID- 12584332 TI - Cellular proteins from human monocytes bind to dengue 4 virus minus-strand 3' untranslated region RNA. AB - The synthesis of plus and minus RNA strands of several RNA viruses requires as a first step the interaction of some viral regulatory sequences with cellular and viral proteins. The dengue 4 virus genome, a single-stranded, positive-polarity RNA, is flanked by two untranslated regions (UTR) located in the 5' and 3' ends. The 3'UTR in the minus-strand RNA [3'UTR (-)] has been thought to function as a promoter for the synthesis of plus-strand RNA. To study the initial interaction between this 3'UTR and cellular and viral proteins, mobility shift assays were performed, and four ribonucleoprotein complexes (I through IV) were formed when uninfected and infected U937 cells (human monocyte cell line) interacted with the 3'UTR (-) of dengue 4 virus. Cross-linking assays with RNAs containing the complete 3'UTR (-) (nucleotides [nt] 101 to 1) or a partial sequence from nt 101 to 45 and nt 44 to 1 resulted in specific binding of some cellular proteins. Supermobility shift and immunoprecipitation assays demonstrated that the La protein forms part of these complexes. To determine the region in the 3' UTR that interacted with the La protein, two deletion mutants were generated. The mutant (del-96), with a deletion of nt 96 to 101, was unable to interact with the La protein, suggesting that La interacted with the 5' portion of the 3'UTR (-). Complex I, which was the main ribonucleoprotein complex formed with the 3'UTR (-) and which had the fastest electrophoretic migration, contained proteins such as calreticulin and protein disulfide isomerase, which constitute important components of the endoplasmic reticulum. PMID- 12584333 TI - Differential impairment of lytic and cytokine functions in senescent human immunodeficiency virus type 1-specific cytotoxic T lymphocytes. AB - Telomere length is abnormally short in the CD8(+) T-cell compartment of human immunodeficiency virus type 1 (HIV-1)-infected persons, likely because of chronic cell turnover. Although clonal exhaustion of CD8(+) cytotoxic T lymphocytes (CTL) has been proposed as a mechanism for loss of antigen-specific responses, the functional consequences of exhaustion are poorly understood. Here we used telomerase transduction to evaluate the impact of senescence on CTL effector functions. Constitutive expression of telomerase in an HIV-1-specific CTL clone results in enhanced proliferative capacity, in agreement with prior studies of other human cell types. Whereas the CTL remain phenotypically normal in terms of antigenic specificity and requirements for proliferation, their cytolytic and antiviral capabilities are superior to those of control CTL. In contrast, their ability to produce gamma interferon and RANTES is essentially unchanged. The selective enhancement of cytolytic function in memory CTL by ectopic telomerase expression implies that loss of this function (but not cytokine production) is a specific consequence of replicative senescence. These data suggest a unifying mechanism for the in vivo observations that telomere lengths are shortened in the CD8(+) cells of HIV-1-infected persons and that HIV-1-specific CTL are deficient in perforin. Telomerase transduction could therefore be a tool with which to explore a potential therapeutic approach to an important pathophysiologic process of immune dysfunction in chronic viral infection. PMID- 12584331 TI - The avian retrovirus avian sarcoma/leukosis virus subtype A reaches the lipid mixing stage of fusion at neutral pH. AB - We previously showed that the envelope glycoprotein (EnvA) of avian sarcoma/leukosis virus subtype A (ASLV-A) binds to liposomes at neutral pH following incubation with its receptor, Tva, at >or=22 degrees C. We also provided evidence that ASLV-C fuses with cells at neutral pH. These findings suggested that receptor binding at neutral pH and >or=22 degrees C is sufficient to activate Env for fusion. A recent study suggested that two steps are necessary to activate avian retroviral Envs: receptor binding at neutral pH, followed by exposure to low pH (W. Mothes et al., Cell 103:679-689, 2000). Therefore, we evaluated the requirements for intact ASLV-A particles to bind to target bilayers and fuse with cells. We found that ASLV-A particles bind stably to liposomes in a receptor- and temperature-dependent manner at neutral pH. Using ASLV-A particles biosynthetically labeled with pyrene, we found that ASLV-A mixes its lipid envelope with cells within 5 to 10 min at 37 degrees C. Lipid mixing was neither inhibited nor enhanced by incubation at low pH. Lipid mixing of ASLV-A was inhibited by a peptide designed to prevent six-helix bundle formation in EnvA; the same peptide inhibits virus infection and EnvA-mediated cell-cell fusion (at both neutral and low pHs). Bafilomycin and dominant-negative dynamin inhibited lipid mixing of Sindbis virus (which requires low pH for fusion), but not of ASLV A, with host cells. Finally, we found that, although EnvA-induced cell-cell fusion is enhanced at low pH, a mutant EnvA that is severely compromised in its ability to support infection still induced massive syncytia at low pH. Our results indicate that receptor binding at neutral pH is sufficient to activate EnvA, such that ASLV-A particles bind hydrophobically to and merge their membranes with target cells. Possible roles for low pH at subsequent stages of viral entry are discussed. PMID- 12584330 TI - Diversity of the human immunodeficiency virus type 1 (HIV-1) env sequence after vertical transmission in mother-child pairs infected with HIV-1 subtype A. AB - Although several virologic and immunologic factors associated with an increased risk of perinatal human immunodeficiency virus type 1 (HIV-1) transmission have been described, the mechanism of mother-to-child transmission is still unclear. More specifically, the question of whether selective pressures influence the transmission remains unanswered. The aim of this study was to assess the genetic diversity of the transmitted virus after in utero transmission and after peripartum transmission and to compare the viral heterogeneity in the child with the viral heterogeneity in the mother. To allow a very accurate characterization of the viral heterogeneity in a single sample, limiting-dilution sequencing of a 1016-bp fragment of the env gene was performed. Thirteen children were tested, including 6 with in utero infections and 7 with peripartum infections. Samples were taken the day after birth and at the ages of 6 and 14 weeks. A homogeneous virus population was seen in six (46.2%) infants, of whom two were infected in utero and four were infected peripartum. A more heterogeneous virus population was detected in seven infants (53.8%), four infected in utero and three infected peripartum. The phylogenetic trees of the mother-child pairs presented a whole range of different tree topologies and showed infection of the child by one or more maternal variants. In conclusion, after HIV-1 transmission from mother to child a heterogeneous virus population was detected in approximately one-half of the children examined. Heterogeneous virus populations were found after peripartum infection as well as after in utero infection. Phylogenetic tree topologies argue against selection processes as the major mechanism driving mother-to-child transmission but support the hypothesis that virus variability is mainly driven by the inoculum level and/or exposure time. PMID- 12584334 TI - Restriction of human immunodeficiency virus type 1 Rev function in murine A9 cells involves the Rev C-terminal domain. AB - The human immunodeficiency virus type 1 (HIV-1) Rev and human T-cell leukemia virus type 1 (HTLV-1) Rex proteins are essential for the expression of viral structural proteins and productive infection. Both contain a nuclear export signal (NES) in their C-terminal domain and a nuclear localization signal (NLS) in their N-terminal domain. The NES and NLS are necessary for shuttling between nucleus and cytoplasm and are therefore indispensable for the transport of unspliced and singly spliced viral transcripts. HIV-1 Rev function is restricted in A9 cells, a murine fibroblast cell line, whereas HTLV-1 Rex is functional in these cells. Immunofluorescence studies with RevGFP fusion protein demonstrate normal import and export of Rev in A9 cells. To ascertain which domains of Rev are necessary for the restriction of Rev function in A9 cells, we studied a chimeric construct in which the NES domain of Rev was exchanged with Rex C terminal amino acids 79 to 95, the Rev1-79/Rex79-95 chimera, which restored Rev function in A9 cells. In addition, overexpression of a truncated Rev containing the Rev C-terminal domain in the presence of wild-type Rev, led to restoration of Rev function in A9 cells. These results suggest that the C-terminal domain of HIV 1 Rev plays an important role in restricting Rev function in murine cells. PMID- 12584335 TI - Origin and evolution of Japanese encephalitis virus in southeast Asia. AB - Since it emerged in Japan in the 1870s, Japanese encephalitis has spread across Asia and has become the most important cause of epidemic encephalitis worldwide. Four genotypes of Japanese encephalitis virus (JEV) are presently recognized (representatives of genotypes I to III have been fully sequenced), but its origin is not known. We have determined the complete nucleotide and amino acid sequence of a genotype IV Indonesian isolate (JKT6468) which represents the oldest lineage, compared it with other fully sequenced genomes, and examined the geographical distribution of all known isolates. JKT6468 was the least similar, with nucleotide divergence ranging from 17.4 to 19.6% and amino acid divergence ranging from 4.7 to 6.5%. It included an unusual series of amino acids at the carboxy terminus of the core protein unlike that seen in other JEV strains. Three signature amino acids in the envelope protein (including E327 Leu-->Thr/Ser on the exposed lateral surface of the putative receptor binding domain) distinguished genotype IV strains from more recent genotypes. Analysis of all 290 JEV isolates for which sequence data are available showed that the Indonesia Malaysia region has all genotypes of JEV circulating, whereas only more recent genotypes circulate in other areas (P < 0.0001). These results suggest that JEV originated from its ancestral virus in the Indonesia-Malaysia region and evolved there into the different genotypes which then spread across Asia. Our data, together with recent evidence on the origins of other emerging viruses, including dengue virus and Nipah virus, imply that tropical southeast Asia may be an important zone for emerging pathogens. PMID- 12584337 TI - Induction of primary virus-cross-reactive human immunodeficiency virus type 1 neutralizing antibodies in small animals by using an alphavirus-derived in vivo expression system. AB - We have studied the induction of neutralizing antibodies by in vivo expression of the human immunodeficiency virus type 1 (HIV-1) envelope by using a Venezuelan equine encephalitis virus (VEE) replicon system with mice and rabbits. The HIV-1 envelope, clone R2, has broad sensitivity to cross-reactive neutralization and was obtained from a donor with broadly cross-reactive, primary virus-neutralizing antibodies (donor of reference serum, HIV-1-neutralizing serum 2 [HNS2]). It was expressed as gp160, as secreted gp140, and as gp160deltaCT with the cytoplasmic tail deleted. gp140 was expressed in vitro at a high level and was predominantly uncleaved oligomer. gp160deltaCT was released by cells in the form of membrane bound vesicles. gp160deltaCT induced stronger neutralizing responses than the other forms. Use of a helper plasmid for replicon particle packaging, in which the VEE envelope gene comprised a wild-type rather than a host range-adapted sequence, also enhanced immunogenicity. Neutralizing activity fractionated with immunoglobulin G. This activity was cross-reactive among a panel of five nonhomologous primary clade B strains and a Chinese clade C strain and minimally reactive against a Chinese clade E (circulating recombinant form 1) strain. The comparative neutralization of these strains by immune mouse sera was similar to the relative neutralizing effects of HNS2, and responses induced in rabbits were similar to those induced in mice. Together, these results demonstrate that neutralizing antibody responses can be induced in mice within 2 to 3 months that are similar in potency and cross-reactivity to those found in the chronically infected, long-term nonprogressive donor of HNS2. These findings support the expectation that induction of highly cross-reactive HIV-1 primary virus neutralizing activity by vaccination may be realized. PMID- 12584336 TI - Simian-human immunodeficiency virus SHIV89.6-induced protection against intravaginal challenge with pathogenic SIVmac239 is independent of the route of immunization and is associated with a combination of cytotoxic T-lymphocyte and alpha interferon responses. AB - Attenuated primate lentivirus vaccines provide the most consistent protection against challenge with pathogenic simian immunodeficiency virus (SIV). Thus, they provide an excellent model to examine the influence of the route of immunization on challenge outcome and to study vaccine-induced protective anti-SIV immune responses. In the present study, rhesus macaques were immunized with live nonpathogenic simian-human immunodeficiency virus (SHIV) 89.6 either intravenously or mucosally (intranasally or intravaginally) and then challenged intravaginally with pathogenic SIVmac239. The route of immunization did not affect mucosal challenge outcome after a prolonged period of systemic infection with the nonpathogenic vaccine virus. Further, protection from the SIV challenge was associated with the induction of multiple host immune effector mechanisms. A comparison of immune responses in vaccinated-protected and vaccinated-unprotected animals revealed that vaccinated-protected animals had higher frequencies of SIV Gag-specific cytotoxic T lymphocytes and gamma interferon (IFN-gamma)-secreting cells during the acute phase postchallenge. Vaccinated-protected animals also had a more pronounced increase in peripheral blood mononuclear cell IFN-alpha mRNA levels than did the vaccinated-unprotected animals in the first few weeks after challenge. Thus, innate as well as cellular anti-SIV immune responses appeared to contribute to the SHIV89.6-induced protection against intravaginal challenge with pathogenic SIVmac239. PMID- 12584338 TI - Human herpesvirus 8 envelope glycoprotein B mediates cell adhesion via its RGD sequence. AB - Human herpesvirus 8 (HHV-8) or Kaposi's sarcoma-associated herpesvirus, implicated in the pathogenesis of Kaposi's sarcoma, utilizes heparan sulfate-like molecules to bind the target cells via its envelope-associated glycoproteins gB and gpK8.1A. HHV-8-gB possesses the Arg-Gly-Asp (RGD) motif, the minimal peptide region of many proteins known to interact with subsets of host cell surface integrins. HHV-8 utilizes alpha3beta1 integrin as one of the receptors for its entry into the target cells via its gB interaction and induces the activation of focal adhesion kinase (FAK) (S. M. Akula, N. P. Pramod, F.-Z. Wang, and B. Chandran, Cell 108:407-419, 2002). Since FAK activation is the first step in the outside-in signaling necessary for integrin-mediated cytoskeletal rearrangements, cell adhesions, motility, and proliferation, the ability of HHV-8-gB to mediate the target cell adhesion was examined. A truncated form of gB without the transmembrane and carboxyl domains (gBdeltaTM) and a gBdeltaTM mutant (gBdeltaTM RGA) with a single amino acid mutation (RGD to RGA) were expressed in a baculovirus system and purified. Radiolabeled HHV-8-gBdeltaTM, gBdeltaTM-RGA, and deltaTMgpK8.1A proteins bound to the human foreskin fibroblasts (HFFs), human dermal microvascular endothelial (HMVEC-d) cells, human B (BJAB) cells, and Chinese hamster ovary (CHO-K1) cells with equal efficiency, which was blocked by preincubation of proteins with soluble heparin. Maxisorp plate-bound gBdeltaTM protein induced the adhesion of HFFs and HMVEC-d and monkey kidney epithelial (CV 1) cells in a dose-dependent manner. In contrast, the gBdeltaTM-RGA and DeltaTMgpK8.1A proteins did not mediate adhesion. Adhesion mediated by gBdeltaTM was blocked by the preincubation of target cells with RGD-containing peptides or by the preincubation of plate-bound gBdeltaTM protein with rabbit antibodies against gB peptide containing the RGD sequence. In contrast, adhesion was not blocked by the preincubation of plate-bound gBdeltaTM protein with heparin, suggesting that the adhesion is mediated by the RGD amino acids of gB, which is independent of the heparin-binding domain of gB. Integrin-ligand interaction is dependent on divalent cations. Adhesion induced by the gBdeltaTM was blocked by EDTA, thus suggesting the role of integrins in the observed adhesions. Focal adhesion components such as FAK and paxillin were activated by the binding of gBdeltaTM protein to the target cells but not by gBdeltaTM-RGA protein binding. Inhibition of FAK phosphorylation by genistein blocked gBdeltaTM-induced FAK activation and cell adhesion. These findings suggest that HHV-8-gB could mediate cell adhesion via its RGD motif interaction with the cell surface integrin molecules and indicate the induction of cellular signaling pathways, which may play roles in the infection of target cells and in Kaposi's sarcoma pathogenesis. PMID- 12584339 TI - Role of quail in the interspecies transmission of H9 influenza A viruses: molecular changes on HA that correspond to adaptation from ducks to chickens. AB - H9 influenza viruses have become endemic in land-based domestic poultry in Asia and have sporadically crossed to pigs and humans. To understand the molecular determinants of their adaptation to land-based birds, we tested the replication and transmission of several 1970s duck H9 viruses in chickens and quail. Quail were more susceptible than chickens to these viruses, and generation of recombinant H9 viruses by reverse genetics showed that changes in the HA gene are sufficient to initiate efficient replication and transmission in quail. Seven amino acid positions on the HA molecule corresponded to adaptation to land-based birds. In quail H9 viruses, the pattern of amino acids at these seven positions is intermediate between those of duck and chicken viruses; this fact may explain the susceptibility of quail to duck H9 viruses. Our findings suggest that quail provide an environment in which the adaptation of influenza viruses from ducks generates novel variants that can cross the species barrier. PMID- 12584340 TI - Tat-neutralizing antibodies in vaccinated macaques. AB - The human immunodeficiency virus Tat protein is essential for virus replication and is a candidate vaccine antigen. Macaques immunized with Tat or chemically modified Tat toxoid having the same clade B sequence developed strong antibody responses. We compared these antisera for their abilities to recognize diverse Tat sequences. An overlapping peptide array covering three clade B and two clade C Tat sequences was constructed to help identify reactive linear epitopes. Sera from Tat-immunized macaques were broadly cross-reactive with clade B and clade C sequences but recognized a clade B-specific epitope in the basic domain. Sera from Tat toxoid-immunized macaques had a more restricted pattern of recognition, reacting mainly with clade B and with only one clade B basic domain sequence, which included the rare amino acids RPPQ at positions 57 to 60. Monoclonal antibodies against the amino terminus or the domain RPPQ sequence blocked Tat uptake into T cells and neutralized Tat in a cell-based transactivation assay. Macaques immunized with Tat or Tat toxoid proteins varied in their responses to minor epitopes, but all developed a strong response to the amino terminus, and antisera were capable of neutralizing Tat in a transactivation assay. PMID- 12584341 TI - As(2)O(3) enhances retroviral reverse transcription and counteracts Ref1 antiviral activity. AB - Potent drugs such as cyclosporine have provided effective probes of signal transduction pathways and, as well, of human immunodeficiency virus type 1 (HIV 1) replication mechanisms. Recently, it was reported that As(2)O(3), a drug used to treat acute promyelocytic leukemia (PML), stimulates HIV-1 replication. We found that As(2)O(3) accelerates the kinetics of a spreading HIV-1 infection in human T cells and increases the number of cells bearing HIV-1 provirus after a single round of infection. The stimulatory effect occurred after membrane fusion and resulted in increased steady-state levels of newly synthesized viral cDNA. Stimulation was independent of HIV-1 env and most viral accessory genes, and As(2)O(3) had no detectable effects on viral expression postintegration or virion assembly. Murine leukemia virus (MLV) transduction was enhanced by As(2)O(3) to the same extent as HIV-1 transduction, but As(2)O(3) had no additional effect on Fv1 restriction. In contrast, As(2)O(3) largely overcame the specific block to N tropic MLV reverse transcription posed by human Ref1. As(2)O(3) disrupts PML bodies, nuclear structures named for a major component, the PML protein. We observed no changes in PML bodies in response to HIV-1 infection. Experiments with PML-null target cells indicated that PML has no effect on HIV-1 infectivity and is dispensable for the stimulatory effect of As(2)O(3). As(2)O(3) caused cell death in uninfected cells at the same concentrations which stimulate HIV-1 replication. Among four additional apoptosis-inducing agents, a boost in HIV-1 infectivity was observed only with carbonyl cyanide m-chlorophenylhydrazone, a compound which, like As(2)O(3), disrupts the mitochondrial transmembrane potential. In summary, As(2)O(3) stimulates retroviral reverse transcription, perhaps via effects on mitochondria, and provides a useful tool for characterizing Ref1. PMID- 12584342 TI - Efficient replication of hepatitis C virus genotype 1a RNAs in cell culture. AB - Hepatitis C virus (HCV) genotype 1 (subtypes 1a and 1b) is responsible for the majority of treatment-resistant liver disease worldwide. Thus far, efficient HCV RNA replication has been observed only for subgenomic and full-length RNAs derived from genotype 1b isolates. Here, we report the establishment of efficient RNA replication systems for genotype 1a strain H77. Replication of subgenomic and full-length H77 1a RNAs required the highly permissive Huh-7.5 hepatoma subline and adaptive amino acid substitutions in both NS3 and NS5A. Replication could be detected by RNA quantification, fluorescence-activated cell sorting, and metabolic labeling of HCV-specific proteins. Replication efficiencies were similar for subgenomic and full-length RNAs and were most efficient for HCV RNAs lacking heterologous RNA elements. Interestingly, both subtype 1a and 1b NS3 adaptive mutations are surface exposed and present on only one face of the NS3 structure. The cell culture-adapted subtype 1a replicons should be useful for basic replication studies and for antiviral development. These results are also encouraging for the development of adapted replicons for the remaining HCV genotypes. PMID- 12584343 TI - Envelopment of human cytomegalovirus occurs by budding into Golgi-derived vacuole compartments positive for gB, Rab 3, trans-golgi network 46, and mannosidase II. AB - Although considerable progress has been made towards characterizing virus assembly processes, assignment of the site of tegumentation and envelopment for human cytomegalovirus (HCMV) is still not clear. In this study, we examined the envelopment of HCMV particles in human lung fibroblasts (HF) HL 411 and HL 19, human umbilical vein endothelial cells, human pulmonary arterial endothelial cells, and arterial smooth muscle cells at different time points after infection by electron microscopy (EM), immunohistochemistry, and confocal microscopy analysis. Double-immunofluorescence labeling experiments demonstrated colocalization of the HCMV glycoprotein B (gB) with the Golgi resident enzyme mannosidase II, the Golgi marker TGN (trans-Golgi network) 46, and the secretory vacuole marker Rab 3 in all cell types investigated. Final envelopment of tegumented capsids was observed at 5 days postinfection by EM, when tegumented capsids budded into subcellular compartments located in the cytoplasm, in close proximity to the Golgi apparatus. Immunogold labeling and EM analysis confirmed staining of the budding compartment with HCMV gB, Rab 3, and mannosidase II in HL 411 cells. However, the markers Rab 1, Rab 2, Rab 7, Lamp 1 (late endosomes and lysosomes), and Lamp 2 (lysosomes) neither showed specific staining of the budding compartment in the immunogold labeling experiments nor colocalized with gB in the immunofluorescent colocalization experiments in any cell type studied. Together, these results suggest that the final envelopment of HCMV particles takes place mainly into a Golgi-derived secretory vacuole destined for the plasma membrane, which may release new infectious virus particles by fusion with the plasma membrane. PMID- 12584344 TI - Subcellular localization of herpes simplex virus type 1 UL51 protein and role of palmitoylation in Golgi apparatus targeting. AB - The herpes simplex virus type 1 (HSV-1) UL51 gene products are virion-associated phosphoproteins with apparent molecular masses of 27, 29, and 30 kDa in HSV-1 infected cells. In this study, we have investigated the intracellular localization and distribution of UL51 protein both in infected cells and in transfected cells expressing only UL51. We found that this protein colocalized closely with Golgi marker proteins such as the Golgi-58K protein and GM130 in transfected cells expressing only UL51. However, in infected cells, the UL51 protein localized to the juxtanuclear region but only partially colocalized with the Golgi maker proteins. Mutant protein analysis revealed that the N-terminal 15 amino acid residues of the UL51 protein sufficed for this Golgi localization property. The UL51 protein redistributed on addition of brefeldin A. This was prevented by pretreatment with 2-deoxyglucose and sodium azide, which results in ATP depletion, but not by pretreatment with NaF and AlCl(3), which activates heterotrimeric G proteins. Moreover, we found that palmitoylation of the UL51 protein through the N-terminal cysteine at position 9 was necessary for its Golgi localization. Protease digestion analysis suggested that the UL51 protein localized on the cytoplasmic face of the membrane in UL51-transfected cells, while in infected cells it localized mainly to the inside of cytoplasmic vesicles and/or the viral envelope. Transmission immunoelectron microscopy revealed an association of UL51 protein-specific labeling with cytoplasmic virions and also with some membranous structure. We infer from these observations that internalization of UL51 protein into the cytoplasmic vesicle and/or virion may occur in association with viral envelopment in HSV-infected cells. PMID- 12584345 TI - An essential role of the enhancer for murine cytomegalovirus in vivo growth and pathogenesis. AB - The transcription of cytomegalovirus (CMV) immediate-early (IE) genes is regulated by a large and complex enhancer containing an array of binding sites for a variety of cellular transcription factors. Previously, using bacterial artificial chromosome recombinants of the virus genome, it was reported that the enhancer region of murine CMV (MCMV) is dispensable but performs a key function for viral multiplication (A. Angulo, M. Messerle, U. H. Koszinowski, and P. Ghazal, J. Virol. 72:8502-8509, 1998). In the present study, we defined, through the reconstitution of infectious enhancerless MCMVs, the growth requirement for the enhancer in tissue culture and explored its significance for steering a productive infection in vivo. A comparison of cis and trans complementation systems for infection of enhancerless virus in permissive fibroblasts revealed a multiplicity-dependent growth phenotype that is severely compromised in the rate of infectious-virus multiplication. The in vivo impact of viruses that have an amputated enhancer was investigated in an extremely sensitive model of MCMV infection, the SCID mouse. Histological examination of spleens, livers, lungs, and salivary glands from animals infected with enhancer-deficient MCMV demonstrated an absence of tissue damage associated with CMV infection. The lack of pathogenic lesions correlated with a defect in replication competence. Enhancerless viruses were not detectable in major target organs harvested from SCID mice. The pathogenesis and growth defect reverted upon restoration of the enhancer. Markedly, while SCID mice infected with 5 PFU of parental MCMV died within 50 days postinfection, all mice infected with enhancerless virus survived for the duration of the experiment (1 year) after infection with 5 x 10(5) PFU. Together, these results clarify the importance of the enhancer for MCMV growth in cell culture and underscore the in vivo significance of this region for MCMV virulence and pathogenesis. PMID- 12584346 TI - Genetic characterization of rebounding human immunodeficiency virus type 1 in plasma during multiple interruptions of highly active antiretroviral therapy. AB - Various strategies of interrupting highly active antiretroviral therapy (HAART) are being investigated for the treatment of human immunodeficiency virus (HIV) infection. Interruptions of greater than 2 weeks frequently result in rebound of plasma HIV RNA. In order to discern changes in the viral population that might occur during cycles of treatment interruption, we evaluated the homology of HIV-1 envelope gene sequences over time in 12 patients who received four to seven cycles of 4 weeks off HAART followed by 8 weeks on HAART by using the heteroduplex tracking assay and novel statistical tools. HIV populations in 9 of 12 patients diverged from those found in the first cycle in at least one subsequent cycle. The substantial genetic changes noted in HIV env did not correlate with increased or decreased log changes in levels of plasma HIV RNA (P > 0.5). Thus, genetic changes in HIV env itself did not contribute in a systematic way to changes in levels of plasma viremia from cycle to cycle of treatment interruption. In addition, the data suggest that there may be multiple compartments contributing to the rebound of plasma viremia and to viral diversity from cycle to cycle of intermittent therapy. PMID- 12584347 TI - Nontransmissible virus-like particle formation by F-deficient sendai virus is temperature sensitive and reduced by mutations in M and HN proteins. AB - The formation of nontransmissible virus-like particles (NTVLP) by cells infected with F-deficient Sendai virus (SeV/deltaF) was found to be temperature sensitive. Analysis by hemagglutination assays and Western blotting demonstrated that the formation of NTVLP at 38 degrees C was about 1/100 of that at 32 degrees C, whereas this temperature-sensitive difference was only moderate in the case of F possessing wild-type SeV. In order to reduce the NTVLP formation with the aim of improving SeV for use as a vector for gene therapy, amino acid substitutions found in temperature-sensitive mutant SeVs were introduced into the M (G69E, T116A, and A183S) and HN (A262T, G264R, and K461G) proteins of SeV/deltaF to generate SeV/M(ts)HN(ts)deltaF. The use of these mutations allows vector production at low temperature (32 degrees C) and therapeutic use at body temperature (37 degrees C) with diminished NTVLP formation. As expected, the formation of NTVLP by SeV/M(ts)HN(ts)deltaF at 37 degrees C was decreased to about 1/10 of that by SeV/deltaF, whereas the suppression of NTVLP formation did not cause either enhanced cytotoxicity or reduced gene expression of the vector. The vectors showed differences with respect to the subcellular distribution of M protein in the infected cells. Clear and accumulated immunocytochemical signals of M protein on the cell surface were not observed in cells infected by SeV/deltaF at an incompatible temperature, 38 degrees C, or in those infected by SeV/M(ts)HN(ts)deltaF at 37 or 38 degrees C. The absence of F protein in SeV/deltaF and the additional mutations in M and HN in SeV/M(ts)HN(ts)deltaF probably weaken the ability to transport M protein to the plasma membrane, leading to the diminished formation of NTVLP. PMID- 12584348 TI - Effects of point mutations in the major capsid protein of beet western yellows virus on capsid formation, virus accumulation, and aphid transmission. AB - Point mutations were introduced into the major capsid protein (P3) of cloned infectious cDNA of the polerovirus beet western yellows virus (BWYV) by manipulation of cloned infectious cDNA. Seven mutations targeted sites on the S domain predicted to lie on the capsid surface. An eighth mutation eliminated two arginine residues in the R domain, which is thought to extend into the capsid interior. The effects of the mutations on virus capsid formation, virus accumulation in protoplasts and plants, and aphid transmission were tested. All of the mutants replicated in protoplasts. The S-domain mutant W166R failed to protect viral RNA from RNase attack, suggesting that this particular mutation interfered with stable capsid formation. The R-domain mutant R7A/R8A protected approximately 90% of the viral RNA strand from RNase, suggesting that lower positive-charge density in the mutant capsid interior interfered with stable packaging of the complete strand into virions. Neither of these mutants systemically infected plants. The six remaining mutants properly packaged viral RNA and could invade Nicotiana clevelandii systemically following agroinfection. Mutant Q121E/N122D was poorly transmitted by aphids, implicating one or both targeted residues in virus-vector interactions. Successful transmission of mutant D172N was accompanied either by reversion to the wild type or by appearance of a second-site mutation, N137D. This finding indicates that D172 is also important for transmission but that the D172N transmission defect can be compensated for by a "reverse" substitution at another site. The results have been used to evaluate possible structural models for the BWYV capsid. PMID- 12584349 TI - Temperature-dependent conformational changes in herpes simplex virus ICP4 that affect transcription activation. AB - The C-terminal 500 amino acids of herpes simplex virus type 1 ICP4 are required for full activator function and viral growth and are known to participate in interactions consistent with the role of ICP4 as an activator of transcription. Oligonucleotide mutagenesis was used to target stretches of amino acids that are conserved with the ICP4 analogs of other alphaherpesviruses and were also predicted to be exposed on the surface of the molecule. Seven mutants were isolated that possessed one to three amino acid changes to the residue alanine in four regions between residues 1000 and 1200. The mutants generated were analyzed first in transfection assays and subsequently after introduction into the viral genome. A number of phenotypes representing different degrees of functional impairment were observed. In transient assays conducted at 37 degrees C, mutant M2 was indistinguishable from wild-type ICP4. Mutants M6 and M7 were marginally impaired. M3, M4, and M5 were more significantly impaired but still able to activate transcription, and M1 was completely impaired. In the context of the viral genome, M1, M3, and M7 were found to be temperature sensitive for growth. All three overproduced immediate-early (IE) proteins at the nonpermissive temperature (NPT). M3 and M7 produced early but not late proteins, and M1 produced neither early nor late proteins, at the NPT. The ICP4 proteins synthesized by all of the mutants tested were able to bind to specific ICP4 binding sites in electrophoretic mobility shift experiments. However, the DNA protein complexes formed with the ICP4 from M1, M3, or M7 produced at the NPT possessed altered mobility. These complexes were not supershifted by a monoclonal antibody that recognizes an epitope in the C terminus; however, they were supershifted by a monoclonal antibody that recognizes the N terminus. The results suggest that the mutant forms of ICP4, while able to bind to DNA, are conformationally altered at the NPT, thus impairing the ability of the protein to activate transcription to different extents. The complete lack of ICP4 function characteristic of the M1 protein, and the inability of all the mutants to attenuate IE gene expression, suggest that the mutations additionally affect functions of the N terminus to different extents. PMID- 12584350 TI - Evaluation of genetically engineered derivatives of a Chinese strain of foot-and mouth disease virus reveals a novel cell-binding site which functions in cell culture and in animals. AB - Adaptation of field isolates of foot-and-mouth disease virus (FMDV) to grow in cells in culture can result in changes in viral properties that include acquisition of the ability to bind to cell surface heparan sulfate (HS). After 13 passages on BHK cells to produce a vaccine, a Cathay topotype isolate of FMDV serotype O from China (O/CHA/90) extended its cell culture host range and bound to heparin-Sepharose, although it did not require cell surface HS as a receptor molecule. To understand these phenomena, we constructed chimeric viruses by using a type A(12) infectious cDNA and the capsid protein-coding regions of O/CHA/90 and its cell culture-adapted derivative (vac-O/CHA/90). Using a set of viruses derived from these chimeras by exchanging portions of the capsid-coding regions, we discovered that a group of amino acid residues that surround the fivefold axis of the icosahedral virion determine host range in cell culture and influence pathogenicity in pigs. These residues included aromatic amino acids at positions 108 and 174 and positively charged residues at positions 83 and 172 in protein 1D. To test if these residues participated in non-integrin-dependent cell binding, the integrin-binding RGD sequence in protein 1D was changed to KGE in two different chimeras. Evaluation of these KGE viruses indicated that growth in cell culture was not dependent on HS. One of these viruses was tested in pigs, where it produced a mild disease and maintained its KGE sequence. These results are discussed in terms of receptor utilization and pathogenesis of this important pathogen. PMID- 12584351 TI - An antiviral peptide targets a coiled-coil domain of the human T-cell leukemia virus envelope glycoprotein. AB - Retrovirus entry into cells is mediated by the viral envelope glycoproteins which, through a cascade of conformational changes, orchestrate fusion of the viral and cellular membranes. In the absence of membrane fusion, viral entry into the host cell cannot occur. For human T-cell leukemia virus type 1 (HTLV-1), synthetic peptides that mimic a carboxy-terminal region of the transmembrane glycoprotein (TM) ectodomain are potent inhibitors of membrane fusion and virus entry. Here, we demonstrate that this class of inhibitor targets a fusion-active structure of HTLV-1 envelope. In particular, the peptides bind specifically to a core coiled-coil domain of envelope, and peptide variants that fail to bind the coiled-coil lack inhibitory activity. Our data indicate that the inhibitory peptides likely function by disrupting the formation of a trimer-of-hairpins structure that is required for membrane fusion. Importantly, we also show that peptides exhibiting dramatically increased potency can be readily obtained. We suggest that peptides or peptide mimetics targeting the fusion-active structures of envelope may be of therapeutic value in the treatment of HTLV-1 infections. PMID- 12584352 TI - Structures of rotavirus reassortants demonstrate correlation of altered conformation of the VP4 spike and expression of unexpected VP4-associated phenotypes. AB - Numerous prior studies have indicated that viable rotavirus reassortants containing structural proteins of heterologous parental origin may express unexpected phenotypes, such as changes in infectivity and immunogenicity. To provide a structural basis for alterations in phenotypic expression, a three dimensional structural analysis of these reassortants was conducted. The structures of the reassortants show that while VP4 generally maintains the parental structure when moved to a heterologous protein background, in certain reassortants, there are subtle alterations in the conformation of VP4. The alterations in VP4 conformation correlated with expression of unexpected VP4 associated phenotypes. Interactions between heterologous VP4 and VP7 in reassortants expressing unexpected phenotypes appeared to induce the conformational alterations seen in VP4. PMID- 12584353 TI - Mumps virus decreases testosterone production and gamma interferon-induced protein 10 secretion by human leydig cells. AB - Mumps virus is responsible for sterility. Here, we show that the mumps virus infects Leydig cells in vitro and totally inhibits testosterone secretion and that ribavirin in mumps virus-infected Leydig cell cultures completely restores testosterone production. Moreover, we show that gamma interferon-induced protein 10 (IP-10) is highly expressed by mumps virus-infected Leydig cells and that ribavirin does not block IP-10 production. PMID- 12584354 TI - Inhibition of gammaherpesvirus replication by RNA interference. AB - RNA interference (RNAi) is a conserved mechanism in which double-stranded, small interfering RNAs (siRNAs) trigger a sequence-specific gene-silencing process. Here we describe the inhibition of murine herpesvirus 68 replication by siRNAs targeted to sequences encoding Rta, an immediate-early protein known as an initiator of the lytic viral gene expression program, and open reading frame 45 (ORF 45), a conserved viral protein. Our results suggest that RNAi can block gammaherpesvirus replication and ORF 45 is required for efficient viral production. PMID- 12584355 TI - Entry of herpes simplex virus type 1 into primary sensory neurons in vitro is mediated by Nectin-1/HveC. AB - Primary cultures of rat and mouse sensory neurons were used to study the entry of herpes simplex virus type 1 (HSV-1). Soluble, truncated nectin-1 but not HveA prevented viral entry. Antibodies against nectin-1 also blocked infection of rat neurons. These results indicate that nectin-1 is the primary receptor for HSV-1 infection of sensory neurons. PMID- 12584356 TI - Role of the 5'-proximal stem-loop structure of the 5' untranslated region in replication and translation of hepatitis C virus RNA. AB - Sequences of the untranslated regions at the 5' and 3' ends (5'UTR and 3'UTR) of the hepatitis C virus (HCV) RNA genome are highly conserved and contain cis acting RNA elements for HCV RNA replication. The HCV 5'UTR consists of two distinct RNA elements, a short 5'-proximal stem-loop RNA element (nucleotides 1 to 43) and a longer element of internal ribosome entry site. To determine the sequence and structural requirements of the 5'-proximal stem-loop RNA element in HCV RNA replication and translation, a mutagenesis analysis was preformed by nucleotide deletions and substitutions. Effects of mutations in the 5'-proximal stem-loop RNA element on HCV RNA replication were determined by using a cell based HCV replicon replication system. Deletion of the first 20 nucleotides from the 5' end resulted in elimination of cell colony formation. Likewise, disruption of the 5'-proximal stem-loop by nucleotide substitutions abolished the ability of HCV RNA to induce cell colony formation. However, restoration of the 5'-proximal stem-loop by compensatory mutations with different nucleotides rescued the ability of the subgenomic HCV RNA to replicate in Huh7 cells. In addition, deletion and nucleotide substitutions of the 5'-proximal stem-loop structure, including the restored stem-loop by compensatory mutations, all resulted in reduction of translation by two- to fivefold, suggesting that the 5'-proximal stem-loop RNA element also modulates HCV RNA translation. These findings demonstrate that the 5'-proximal stem-loop of the HCV RNA is a cis-acting RNA element that regulates HCV RNA replication and translation. PMID- 12584357 TI - Reversible oxidative modification as a mechanism for regulating retroviral protease dimerization and activation. AB - Human immunodeficiency virus protease activity can be regulated by reversible oxidation of a sulfur-containing amino acid at the dimer interface. We show here that oxidation of this amino acid in human immunodeficiency virus type 1 protease prevents dimer formation. Moreover, we show that human T-cell leukemia virus type 1 protease can be similarly regulated through reversible glutathionylation of its two conserved cysteine residues. Based on the known three-dimensional structures and multiple sequence alignments of retroviral proteases, it is predicted that the majority of retroviral proteases have sulfur-containing amino acids at the dimer interface. The regulation of protease activity by the modification of a sulfur-containing amino acid at the dimer interface may be a conserved mechanism among the majority of retroviruses. PMID- 12584358 TI - Duplication of U3 sequences in the long terminal repeat of mink cell focus inducing viruses generates redundancies of transcription factor binding sites important for the induction of thymomas. AB - The ability of mink cell focus-inducing (MCF) viruses to induce thymomas is determined, in part, by transcriptional enhancers in the U3 region of their long terminal repeats (LTRs). To elucidate sequence motifs important for enhancer function in vivo, we injected newborn mice with MCF 1dr (supF), a weakly pathogenic, molecularly tagged (supF) MCF virus containing only one copy of a sequence that is present as two copies (known as the directly repeated [DR] sequence) in the U3 region of MCF 247 and analyzed LTRs from supF-tagged proviruses in two resulting thymomas. Tagged proviruses integrated upstream and in the reverse transcriptional orientation relative to c-myc provided the focus of our studies. These proviruses are thought to contribute to thymoma induction by enhancer-mediated deregulation of c-myc expression. The U3 region in a tagged LTR in one thymoma was cloned and sequenced. Relative to MCF 1dr (supF), the cloned U3 region contained an insertion of 140 bp derived predominantly from the DR sequence of the injected virus. The inserted sequence contains predicted binding sites for transcription factors known to regulate the U3 regions of various murine leukemia viruses. Similar constellations of binding sites were duplicated in two proviral LTRs integrated upstream from c-myc in a second thymoma. We replaced the U3 sequences in an infectious molecular clone of MCF 247 with the cloned proviral U3 sequences from the first thymoma and generated an infectious chimeric virus, MCF ProEn. When injected into neonatal AKR mice, MCF ProEn was more pathogenic than the parental virus, MCF 1dr (supF), as evidenced by the more rapid onset and higher incidence of thymomas. Molecular analyses of the resultant thymomas indicated that the U3 region of MCF ProEn was genetically stable. These data suggest that the arrangement and/or redundancy of transcription factor binding sites generated by specific U3 sequence duplications are important to the biological events mediated by MCF proviruses integrated near c-myc that contribute to transformation. PMID- 12584359 TI - Ebola virus transcription activator VP30 is a zinc-binding protein. AB - Ebola virus VP30 is an essential activator of viral transcription. In viral particles, VP30 is closely associated with the nucleocapsid complex. A conspicuous structural feature of VP30 is an unconventional zinc-binding Cys(3) His motif comprising amino acids 68 to 95. By using a colorimetric zinc-binding assay we found that the VP30-specific Cys(3)-His motif stoichiometrically binds zinc ions in a one-to-one relationship. Substitution of the conserved cysteines and the histidine within the motif led to a complete loss of the capacity for zinc binding. Functional analyses revealed that none of the tested mutations of the proposed zinc-coordinating residues influenced binding of VP30 to nucleocapsid-like particles but, concerning its role in activating viral transcription, all resulted in a protein that was inactive. PMID- 12584360 TI - Defects in virion production caused by mutations affecting the C-terminal portion of the Moloney murine leukemia virus capsid protein. AB - The capsid (CA) domain of the Moloney murine leukemia virus (Mo-MuLV) Gag protein has a unique carboxy terminus with a highly charged arginine-rich sequence. Mutant viruses harboring arginine-to-alanine mutations affecting this region of CA displayed significant defects in virion release, and the few viral particles produced were noninfectious. The interaction between the mutant Gag precursors was affected, as judged by the yeast two-hybrid assay. The results suggest that the unique carboxy terminus of CA in the Mo-MuLV plays an important role in Gag Gag association during virion production. PMID- 12584362 TI - Assessment of genetic susceptibility to ethanol intoxication in mice. AB - Increased use of gene manipulation in mice (e.g., targeted or random mutagenesis) has been accompanied by increased reliance on a very few rapid and simple behavioral assays, each of which aspires to model a human behavioral domain. Yet, each assay comprises multiple traits, influenced by multiple genetic factors. Motor incoordination (ataxia), a common characteristic of many neurological disorders, may reflect disordered balance, muscle strength, proprioception, and/or patterned gait. Impaired motor performance can confound interpretation of behavioral assays of learning and memory, exploration, motivation, and sensory competence. The rotarod is one of the most commonly used tests to measure coordination in mice. We show here that exactly how the rotarod test is performed can markedly alter the apparent patterns of genetic influence both in undrugged performance and sensitivity to ethanol intoxication. However, when tested with well chosen parameters, the accelerating rotarod test showed very high inter- and intralaboratory reliability. Depending on test conditions, ethanol can either disrupt or enhance performance in some strains. Genetic contribution to performance on the accelerating versus the fixed-speed rotarod assay can be completely dissociated under some test conditions, and multiple test parameters are needed to assess the range of genetic influence adequately. PMID- 12584361 TI - Replacement of murine leukemia virus readthrough mechanism by human immunodeficiency virus frameshift allows synthesis of viral proteins and virus replication. AB - Retroviruses use unusual recoding strategies to synthesize the Gag-Pol polyprotein precursor of viral enzymes. In human immunodeficiency virus, ribosomes translating full-length viral RNA can shift back by 1 nucleotide at a specific site defined by the presence of both a slippery sequence and a downstream stimulatory element made of an extensive secondary structure. This so called frameshift mechanism could become a target for the development of novel antiviral strategies. A different recoding strategy is used by other retroviruses, such as murine leukemia viruses, to synthesize the Gag-Pol precursor; in this case, a stop codon is suppressed in a readthrough process, again due to the presence of a specific structure adopted by the mRNA. Development of antiframeshift agents will greatly benefit from the availability of a simple animal and virus model. For this purpose, the murine leukemia virus readthrough region was rendered inactive by mutagenesis and the frameshift region of human immunodeficiency virus was inserted to generate a chimeric provirus. This substitution of readthrough by frameshift allows the synthesis of viral proteins, and the chimeric provirus sequence was found to generate infectious viruses. This system could be a most interesting alternative to study ribosomal frameshift in the context of a virus amenable to the use of a simple animal model. PMID- 12584363 TI - Kinetics of size changes of individual Bacillus thuringiensis spores in response to changes in relative humidity. AB - Using an automated scanning microscope, we report the surprising result that individual dormant spores of Bacillus thuringiensis grow and shrink in response to increasing and decreasing relative humidity. We simultaneously monitored the size of inorganic calibration particles. We found that the spores consistently swell in response to increased relative humidity, and shrink to near their original size on reexposure to dry air. Although the dispersion of swelling amplitudes within an ensemble of spores is wide (approximately 30% of the average amplitude), amplitudes for individual spores are highly correlated between different swelling episodes, suggesting that individual spores respond consistently to changes in humidity. We find evidence for two distinct time scales for swelling: one with a time scale of no more than approximately 50 s, and another with a time scale of approximately 8 min. We speculate that these two mechanisms may be due to rapid diffusion of water into the spore coat + cortex, followed by slower diffusion of water into the spore core, respectively. Humidity dependent swelling may account for the greater kill effectiveness of spores by gas-phase chlorine dioxide, formaldehyde, and ethylene oxide at very high relative humidity. PMID- 12584364 TI - Rethinking postmenopausal hormone therapy. PMID- 12584365 TI - Endothelial progenitor cells. PMID- 12584366 TI - A comparison of outcomes with angiotensin-converting--enzyme inhibitors and diuretics for hypertension in the elderly. AB - BACKGROUND: Treatment of hypertension with diuretics, beta-blockers, or both leads to improved outcomes. It has been postulated that agents that inhibit the renin-angiotensin system confer benefit beyond the reduction of blood pressure alone. We compared the outcomes in older subjects with hypertension who were treated with angiotensin-converting-enzyme (ACE) inhibitors with the outcomes in those treated with diuretic agents. METHODS: We conducted a prospective, randomized, open-label study with blinded assessment of end points in 6083 subjects with hypertension who were 65 to 84 years of age and received health care at 1594 family practices. Subjects were followed for a median of 4.1 years, and the total numbers of cardiovascular events in the two treatment groups were compared with the use of multivariate proportional-hazards models. RESULTS: At base line, the treatment groups were well matched in terms of age, sex, and blood pressure. By the end of the study, blood pressure had decreased to a similar extent in both groups (a decrease of 26/12 mm Hg). There were 695 cardiovascular events or deaths from any cause in the ACE-inhibitor group (56.1 per 1000 patient years) and 736 cardiovascular events or deaths from any cause in the diuretic group (59.8 per 1000 patient-years; the hazard ratio for a cardiovascular event or death with ACE-inhibitor treatment was 0.89 [95 percent confidence interval, 0.79 to 1.00]; P=0.05). Among male subjects, the hazard ratio was 0.83 (95 percent confidence interval, 0.71 to 0.97; P=0.02); among female subjects, the hazard ratio was 1.00 (95 percent confidence interval, 0.83 to 1.21; P=0.98); the P value for the interaction between sex and treatment-group assignment was 0.15. The rates of nonfatal cardiovascular events and myocardial infarctions decreased with ACE-inhibitor treatment, whereas a similar number of strokes occurred in each group (although there were more fatal strokes in the ACE-inhibitor group). CONCLUSIONS: Initiation of antihypertensive treatment involving ACE inhibitors in older subjects, particularly men, appears to lead to better outcomes than treatment with diuretic agents, despite similar reductions of blood pressure. PMID- 12584367 TI - Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. AB - BACKGROUND: Cardiovascular risk factors contribute to atherogenesis by inducing endothelial-cell injury and dysfunction. We hypothesized that endothelial progenitor cells derived from bone marrow have a role in ongoing endothelial repair and that impaired mobilization or depletion of these cells contributes to endothelial dysfunction and cardiovascular disease progression. METHODS: We measured the number of colony-forming units of endothelial progenitor cells in peripheral-blood samples from 45 men (mean [+/-SE] age, 50+/-2 years). The subjects had various degrees of cardiovascular risk but no history of cardiovascular disease. Endothelium-dependent and endothelium-independent function was assessed by high-resolution ultrasonography of the brachial artery. RESULTS: We observed a strong correlation between the number of circulating endothelial progenitor cells and the subjects' combined Framingham risk factor score (r=-0.47, P=0.001). Measurement of flow-mediated brachial-artery reactivity also revealed a significant relation between endothelial function and the number of progenitor cells (r=0.59, P<0.001). Indeed, the levels of circulating endothelial progenitor cells were a better predictor of vascular reactivity than was the presence or absence of conventional risk factors. In addition, endothelial progenitor cells from subjects at high risk for cardiovascular events had higher rates of in vitro senescence than cells from subjects at low risk. CONCLUSIONS: In healthy men, levels of endothelial progenitor cells may be a surrogate biologic marker for vascular function and cumulative cardiovascular risk. These findings suggest that endothelial injury in the absence of sufficient circulating progenitor cells may affect the progression of cardiovascular disease. PMID- 12584368 TI - Influence of immunogenicity on the long-term efficacy of infliximab in Crohn's disease. AB - BACKGROUND: Treatment with infliximab, a chimeric monoclonal IgG1 antibody against tumor necrosis factor, can result in the formation of antibodies against infliximab. We evaluated the clinical significance of these antibodies in patients with Crohn's disease. METHODS: In a cohort of 125 consecutive patients with Crohn's disease who were treated with infliximab infusions, we evaluated the concentrations of infliximab and of antibodies against infliximab, clinical data, side effects (including infusion reactions), and the use of concomitant medications before and 4, 8, and 12 weeks after each infusion. RESULTS: A mean of 3.9 infusions (range, 1 to 17) per patient were administered over a mean period of 10 months. Antibodies against infliximab were detected in 61 percent of patients. The presence of concentrations of 8.0 microg per milliliter or greater before an infusion predicted a shorter duration of response (35 days, as compared with 71 days among patients with concentrations of less than 8.0 microg per milliliter; P<0.001) and a higher risk of infusion reactions (relative risk, 2.40; 95 percent confidence interval, 1.65 to 3.66; P<0.001). Infliximab concentrations were significantly lower at four weeks among patients who had had an infusion reaction than among patients who had never had an infusion reaction (median, 1.2 vs. 14.1 microg per milliliter; P<0.001). Patients who had infusion reactions had a median duration of clinical response of 38.5 days, as compared with 65 days among patients who did not have an infusion reaction (P<0.001). Concomitant immunosuppressive therapy was predictive of low titers of antibodies against infliximab (P<0.001) and high concentrations of infliximab four weeks after an infusion (P<0.001). CONCLUSIONS: The development of antibodies against infliximab is associated with an increased risk of infusion reactions and a reduced duration of response to treatment. Concomitant immunosuppressive therapy reduces the magnitude of the immunogenic response. PMID- 12584369 TI - Images in clinical medicine. Cardiac rupture. PMID- 12584370 TI - Initial treatment of hypertension. PMID- 12584371 TI - Selective estrogen-receptor modulators -- mechanisms of action and application to clinical practice. PMID- 12584372 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 5-2003 - a 16-year-old girl with a rash and chest pain. PMID- 12584373 TI - Treating hypertension - what are we to believe? PMID- 12584375 TI - In search of a better HIV vaccine - the heat is on. PMID- 12584376 TI - Understanding the divergent data on postmenopausal hormone therapy. PMID- 12584377 TI - Infection control - a problem for patient safety. PMID- 12584378 TI - Breast-conserving surgery for breast cancer. PMID- 12584379 TI - Heart failure - incidence and survival. PMID- 12584380 TI - Digoxin for the treatment of heart failure. PMID- 12584381 TI - State expenditures for tobacco-control programs and the tobacco settlement. PMID- 12584382 TI - Residents' work hours. PMID- 12584383 TI - Review of heat wave: social autopsy of disaster in Chicago. PMID- 12584384 TI - Ectopic ovary and unicornuate uterus. PMID- 12584385 TI - A rare case of serratia sepsis and spontaneous abortion. PMID- 12584386 TI - Advances in the diagnosis and treatment of pulmonary embolism. Pulmonary embolism -how can you mend a broken clot? PMID- 12584387 TI - Principle mechanisms underlying venous thromboembolism: epidemiology, risk factors, pathophysiology and pathogenesis. AB - Deep vein thrombosis (DVT) and pulmonary embolism (PE) are distinct but related aspects of the same dynamic disease process known as venous thromboembolism (VTE). An estimated 200,000 new cases occur in the United States every year, including 94,000 with PE, resulting in an incidence of 23 per 100,000 patients per year-cases. Without treatment, pulmonary embolism is associated with a mortality rate of approximately 30%, causing nearly 50,000 deaths per year. Moreover, based on post-mortem studies, two-thirds of the patients with pulmonary emboli remain undiagnosed. Clinically, PE may present as (1) isolated dyspnea, (2) pleuritic pain and/or hemoptysis, and (3) circulatory collapse. However, clinical history and examination can be notoriously misleading in reaching a diagnosis. A number of acquired etiologic risk factors (predispositions) are associated with a tendency to develop VTE. These include increasing age, immobilization, surgery, trauma, hospital or nursing home confinement, malignancy, neurologic disease with extremity paresis, as well as certain types of oral contraception and hormone replacement therapy. In addition, a variety of genetic risk factors, such as factor V Leiden, protein S or C deficiency have also been identified. However, in at least half of the instances, no predisposing factors can be identified (idiopathic PE). In the majority of cases thromboemboli originate in the deep veins of the calf or pelvis. The pathogenic conditions for VTE comprise a triad of factors and include (1) venous stasis, (2) hypercoagulable states, and (3) vascular endothelium injury. Occlusion of pulmonary arteries has variable and transient clinical and pathophysiologic consequences, involving both mechanical and reflex effects of vascular occlusion with a consecutive perfusion defect as well as the release of vasoactive and other inflammatory mediators. The objectives of this article are to present an overview of the etiologic and pathogenic factors promoting VTE as well as the pathophysiologic and inflammatory processes following PE. PMID- 12584388 TI - Predicting chemotherapy response to paclitaxel-based therapy in advanced non small-cell lung cancer with P-glycoprotein expression. AB - BACKGROUND: It was reported that multidrug resistance gene 1 (MDR1) encoding human P-glycoprotein (Pgp) may play an important role in multidrug resistance of lung cancer. Therefore, before initiating chemotherapy, it is important to accurately determine the presence of Pgp in lung cancer, to achieve a satisfactory chemotherapy response. OBJECTIVES: The aim of this study was to compare immunohistochemical analyses of Pgp expression and response to paclitaxel in non-small-cell lung cancer (NSCLC). METHODS: Before chemotherapy with paclitaxel, 50 patients with stage IIIb or IV NSCLC were enrolled in this study. Immunohistochemical analyses were performed on multiple nonconsecutive sections of the biopsy specimens to determine Pgp expression. Chemotherapy response was evaluated in the 3rd month after completion of treatment by clinical and radiological methods. RESULTS: All of the 28 (100%) cases with good response had negative Pgp expression and 15 of the 22 (68%) cases with poor response had positive Pgp expression (p < 0.05). No significant differences were found for other prognostic factors (age, sex, body weight loss, performance status, tumor cell type, and tumor stage) between good response and poor response groups. CONCLUSIONS: Although Pgp expression in NSCLC does not fully predict chemotherapy response to paclitaxel-based therapy, detection of Pgp expression will aid in planning paclitaxel-based therapy for patients with advanced NSCLC. Further studies with a larger number of patients and a longer time of follow-up are necessary to confirm our findings. PMID- 12584389 TI - High-resolution CT features: prognostic significance in peripheral lung adenocarcinoma with bronchioloalveolar carcinoma components. AB - BACKGROUND: Based on Noguchi's classification, adenocarcinomas with bronchioloalveolar carcinoma (BAC) components have a heterogeneous prognosis. However, until now, the prognostic factors in this tumor category have not been clarified. OBJECTIVES: We studied the prognostic significance of high-resolution CT (HRCT) findings in this tumor subtype. MATERIALS AND METHODS: HRCT findings [lesion size, percentage of ground-glass opacity (GGO) areas in the lesion, presence or absence of lobulation, coarse spiculation, pleural tag, satellite lesion, air bronchogram, and site of the lesion], clinical data (age, gender and surgical method), and pathologic findings (degree of tumor differentiation and presence or absence of nodal metastasis) were analyzed in 52 consecutive patients with adenocarcinoma with BAC components <3 cm (mean, 1.7 cm). The patients included 32 women and 20 men with a mean age of 66 years. The results correlated with survival using univariate and multivariate analysis. The mean follow-up period was 41 months. RESULTS: Nine (17%) of the 52 patients died of lung cancer. In univariate analysis, a significant difference was found for lobulation (p = 0.018), %GGO (p = 0.043), air bronchogram (p = 0.003), site of lesion (p = 0.034), degree of tumor differentiation (p = 0.001), and nodal metastasis (p = 0.040). Multivariate analysis using the 6 factors that were significant in univariate analysis as independent variables revealed that an air bronchogram (p = 0.010) and the degree of tumor differentiation (p = 0.008) were significant independent factors for survival. CONCLUSION: The presence of an air bronchogram on HRCT may have an independent prognostic significance in lung adenocarcinoma with BAC components. PMID- 12584390 TI - Immune cells in bronchoalveolar lavage in peripheral lung cancer--analysis of 140 cases. AB - BACKGROUND: Changes in the bronchoalveolar lavage fluid (BALF) cell profile may reflect immunologic reactions of the lung in pulmonary malignancies. OBJECTIVE: Our aim was to analyse the BALF cell profile in peripheral lung cancer. METHODS: This study was a retrospective analysis of BALF samples containing tumor cells from patients with confirmed peripheral lung cancer (140 cases). Routine analysis and lymphocyte phenotyping by an immunoperoxidase technique with the monoclonal antibodies CD3, CD20, CD4, CD8, CD57, CD1, CD25, CD71 and HLA-DR were performed. The data were compared with those from healthy persons. RESULTS: We found a significantly lower proportion of macrophages (60%), and significantly elevated proportions of lymphocytes (24%) and neutrophils (13%) in cases with cancer when compared with controls. The proportion of eosinophils was higher in the cancer patients (2.4%), though not significantly. Regarding the lymphocyte phenotypes, we found a significantly elevated proportion of T cells (86%), a relatively low CD4/CD8 ratio (1.7 +/- 1.3), and a relatively higher percentage of HLA-DR+ lymphocytes (6.6%) in patients with cancer. There were no significant differences between smokers and nonsmokers in the cancer patients. In more than half of the cases, a positive staining for epithelial membrane antigen was found. CONCLUSION: Our observations confirm the possible participation of lymphocytes (activated and suppressor subtypes) and eosinophils in the response against tumor in peripheral airways. PMID- 12584391 TI - Substance P-immunoreactive nerves in endobronchial biopsies in cough-variant asthma and classic asthma. AB - BACKGROUND: Unlike classic asthma, cough-variant asthma does not show any evidence of airway obstruction. The main symptom is a dry cough with little known pathophysiology. Hypersensitivity of the cough receptors in cough-variant asthma and an increase in the sensory nerve density of the airway epithelium in persistent dry cough patients have been reported. Therefore, it is possible that there is a higher sensory nerve density in cough-variant asthma patients than in classic asthma patients. OBJECTIVES: This study was undertaken to compare the substance P (SP)-immunoreactive nerve density in mucosal biopsies of cough variant asthma patients, classic asthma patients, and in control subjects. METHODS: Bronchoscopic biopsies were performed in 6 cough-variant asthma patients, 14 classic asthma patients, and 5 normal controls. The tissues obtained were stained immunohistochemically. The SP-immunoreactive nerve density was measured in the bronchial epithelium using a light microscope at 400 x magnification. RESULTS: SP- immunoreactive nerve density for the cough-variant asthma group was significantly higher than that of the classic asthma group (p = 0.001), and of the normal control group (p = 0.006). CONCLUSIONS: It is possible that a sensory nerve abnormality within the airway may be related to hypersensitivity of the cough receptor, and that this may be one of the pathophysiologies of cough-variant asthma. PMID- 12584392 TI - Influence of predicted FEV1 on bronchodilator response in asthmatic patients. AB - BACKGROUND: There is currently disagreement on the way of expressing the reversibility of airflow obstruction, with some evaluations based on the initial FEV(1) while others use predicted FEV(1) (according to age, gender and height). OBJECTIVES: To test the relevance of expressing bronchodilator response as a percentage of predicted FEV(1), we evaluated the influence of morphological data on the response to bronchodilators in a population with a large range of predicted values. METHODS: We measured the change in FEV(1) after inhalation of 200 microg of salbutamol in 30 asthmatic subjects (15 adults and 15 children) in whom predicted FEV(1) ranged between 1.13 and 4.10 liters and analyzed the respective influence of initial and predicted FEV(1 )on bronchodilator response. RESULTS: We have shown a significant relationship between the absolute variation in FEV(1), in liters, and predicted FEV(1) (p = 0.0019). There was also a significant relationship between the absolute variation in FEV(1) and initial FEV(1), in liters (p = 0.02). This relationship was no longer significant (p = 0.8) when the variation and initial FEV(1) were both expressed as percentages of predicted FEV(1). In addition, multiple regression analysis showed that predicted FEV(1) was the only independent variable correlating with the response to bronchodilators. CONCLUSION: This study showed the influence of predicted FEV(1) on bronchodilator response. This result provides an additional argument for expressing bronchodilator response as a percentage of predicted FEV(1). PMID- 12584393 TI - Bronchial hyperreactivity and allergic status in inflammatory bowel disease. AB - BACKGROUND: Despite the known systemic manifestations of inflammatory bowel disease (IBD) and a large number of reports associating lung disease and IBD, the frequency of atopy and bronchial hyperreactivity (BHR) in IBD remains obscure. OBJECTIVES: The aim of this study was to investigate the prevalence of abnormal pulmonary function tests, BHR and the atopic status in patients with IBD. METHODS: Thirty patients with IBD (19 with ulcerative colitis and 11 with Crohn's disease; 19 male, 11 female) and 16 controls without any gastrointestinal disease (9 female, 7 male) were included. Patients were questioned with respect to pulmonary and allergic symptoms; subsequently, lung function tests, BHR, skin prick test positivity, peripheral eosinophilia and serum IgE levels were evaluated and compared with those of control subjects. RESULTS: The mean duration of IBD was 5.3 +/- 4.8 years. IBD patients had significantly more often respiratory symptoms in comparison with controls (odds ratio, OR: 9.0, p < 0.04). A previous diagnosis of asthma and antiasthmatic drug treatment were noted in 3/30 (10%) IBD patients. Allergic symptoms were more prevalent in IBD patients in comparison with the controls (OR: 13, p < 0.007), particularly in patients with ulcerative colitis (OR: 16, p < 0.004). The mean FEV(1 )was 3.1 +/- 0.9 liters (96 +/- 18% predicted), mean methacholine PD(20): 14.7 +/- 3.6 mg/ml, mean IgE: 190.5 +/- 305.6 IU/ml (normal value <94 IU/ml) and the percentage of peripheral eosinophils was 3.1 +/- 3.3% in the IBD patients. These values did not result in statistically significant differences in comparison with controls. Furthermore, abnormal lung function and BHR were observed in 8/30 (27%) and 5/30 (17%) IBD patients, respectively. Abnormal lung function tests were more prevalent in the IBD patients than in the controls (OR: 12, p < 0.04). Skin prick tests were positive in 15/30 (50%) IBD patients. The risk of a positive skin prick test increased in the IBD patients in comparison with the controls (OR: 7.0, p < 0.02). Duration and activity of IBD did not influence the prevalence of BHR, allergic and respiratory symptoms, abnormal lung function, high serum IgE levels and skin test positivity. CONCLUSIONS: Allergic symptoms, respiratory symptoms, abnormal lung function tests and skin prick test positivity were more common among the IBD patients in comparison with the controls. PMID- 12584394 TI - Newly diagnosed chronic obstructive pulmonary disease. Clinical features and distribution of the novel stages of the Global Initiative for Obstructive Lung Disease. AB - BACKGROUND: The new guidelines of the Global Initiative for Obstructive Lung Disease (GOLD) propose a novel staging system for COPD. This study describes the frequency distribution of GOLD stages in newly diagnosed COPD patients in a large city pulmonary practice. METHODS: All patients newly admitted between 1995 and 1996 were analyzed retrospectively. Incident COPD cases were classified according to GOLD criteria. RESULTS: Among 1,434 patients, 210 were diagnosed with chronic obstructive pulmonary disease (COPD) (60% males, age 55 years, range 20-82 years). 67.5% of the patients were current smokers, 27% ex-smokers, and 5.5% nonsmokers. Based on GOLD criteria, 37% had stage 0, 5% stage I, 46% stage II, and 12% stage III COPD. Symptoms leading patients to seek medical advice were cough (84%), exertional dyspnea (70%), and sputum (45%), with a median symptom duration of 12 months (range 1-240 months). Compared with patients with GOLD stages 0-1, those with stages 2-3 were older (60 vs. 47 years, p < 0.001), heavier smokers (40 vs. 20 pack-years, p < 0.001), had a longer duration of symptoms (24 vs. 6 months, p < 0.001), and elevated IgE (stage 3 only, p < 0.04 vs. stages 0-2). Interestingly, stage 0 COPD patients did not have 'normal' spirometry, as indicated by significantly lower FEV(1) (% predicted) and FEF(25 75) (% predicted), compared with age-matched nonsmoking controls (93.1 +/- 1.8 vs. 99 +/- 1.6, p = 0.004; and 76.2 +/- 2.8 vs. 91.2 +/- 2.9, p = 0.0003, respectively). CONCLUSIONS: The majority of COPD patients seek medical advice at advanced disease stages, and smoke actively despite severe symptoms and functional impairment. However, nearly every second patient presents at stages 0 1, thus opening a window for therapeutic or behavioral intervention. GOLD guidelines are a useful basis to reinforce screening programs aimed at early detection and prevention of progressive COPD in individuals at risk and smoking cessation. PMID- 12584395 TI - Analysis of chest CT in patients with Mycobacterium avium complex pulmonary disease. AB - BACKGROUND: The radiographic changes of Mycobacterium avium complex (MAC) pulmonary disease during therapy have not been studied well. OBJECTIVE: To assess the efficacy of antituberculous drug therapy against MAC pulmonary disease using computed tomography (CT). METHOD: We analyzed chest CT scans before and after antituberculous therapy in 30 patients (21 women, 9 men) with MAC pulmonary disease. To evaluate radiographic changes during therapy, we defined a 'degree of improvement' (DI) that is calculated according to the CT appearance. RESULTS: DI was better (1.35 +/- 0.21) in patients who had converted sputum culture than in those who had not (0.44 +/- 0.25) (p < 0.05). In patients who were diagnosed by bronchial washing, DI was better (1.60 +/- 0.22) than in patients who were diagnosed by sputum (0.67 +/- 0.20) (p < 0.01). We categorized the CT appearance into 6 types: small nodules, cavities, bronchial wall thickening, infiltration, pleural thickening and atelectasis. Patients who showed pleural thickening had a significantly worse DI (0.12 +/- 0.40) than those who did not (1.23 +/- 0.18) (p < 0.01). Most of the lesions that disappeared after therapy were small nodules. CONCLUSION: These results indicate that chest CT might be a useful tool for the prediction or assessment of drug therapy for MAC pulmonary disease. PMID- 12584396 TI - Repeated thoracentesis: an important risk factor for eosinophilic pleural effusion? AB - BACKGROUND: Eosinophilic pleural effusion (EPE) is a relatively rare clinical condition. Repeated thoracenteses (RTs) are normally considered a frequent cause of EPE. Yet, to our knowledge, there is no firm evidence (apart from anecdotal case reports) supporting such a statement. OBJECTIVE: To investigate potential relationships between the number, type (with or without pleural biopsy) and time elapsed between RTs and the number of eosinophils present in pleural fluid samples. METHODS: We reviewed retrospectively 273 pleural fluid samples belonging to 120 patients (79 males, 41 females), attended in our institution from 1992 to 2000, whose clinical management had required RTs. Apart from the anthropometric and clinical data of each patient, we included the following variables in the analysis: number of thoracenteses performed in each individual, number of pleural biopsies carried out at each thoracentesis and time between consecutive thoracenteses. We also recorded the total (and differential) leukocyte count, red blood cell count, as well as the main biochemical, microbiological and histological data of both the pleural fluid and peripheral blood samples. RESULTS: We did not observe any significant change in the percentage of eosinophils in relation to the number of thoracenteses performed per patient. This lack of relationship was also observed in the subgroup of patients who required one (or more) pleural biopsies (n = 111) (regardless of the number of biopsies). Our results suggest that RTs are not an important risk factor for the development of EPE, regardless of the time elapsed between two thoracenteses. CONCLUSION: We believe, therefore, that multiple punctures should not longer be considered a prevalent cause of pleural eosinophilia. PMID- 12584397 TI - Transthoracic ultrasound. AB - Sonography (US) has inherent limitations for thoracic imaging because sound waves are reflected by bone and air space (such as in lung parenchyma). However, US is less expensive and more convenient than computed tomography (CT) or magnetic resonance imaging (MRI); it provides immediate information with real-time imaging, and can give information not available from a standard radiograph. This review describes the utility and limitations of US and compares US with radiography. PMID- 12584398 TI - Neuronal nitric oxide synthase is associated with airway obstruction in BALB/c mice exposed to ozone. AB - BACKGROUND: The functional role of nitric oxide (NO) and the various nitric oxide synthase (NOS) isoforms in asthma is controversial. OBJECTIVE: To investigate the role of NO in mice exposed to ozone, three known isoforms of NOS [inducible NOS (iNOS), neuronal NOS (nNOS), and endothelial NOS (eNOS)] were studied. METHODS: The expression of iNOS, nNOS, and eNOS was determined in lung by Western blot analysis after exposure to filtered air and ozone (0.12, 0.5, 1 or 2 ppm) for 3 h. Using barometric whole-body plethysmography and increase in enhanced pause (P(enh)) as an index of airway obstruction, we measured airway responses to ozone exposure. Bronchoalveolar lavage (BAL) was performed. Nitrate and nitrite were measured using a modified Griess reaction. RESULTS: The nitrate concentration in BAL fluid, which indicates the in vivo generation of NO in airways, from the ozone-exposed group was significantly greater than that from the group exposed to filtered air (631.0 +/- 86.4 vs. 152.1 +/- 16.9 micromol/l, p < 0.05). The nitrate concentration in BAL fluid was increased more in mice exposed to 2-ppm ozone than that in mice exposed to filtered air or 0.12-, 0.5-, or 1-ppm ozone. Increases in P(enh) after exposure to ozone or filtered air were significantly higher in the ozone-exposed groups than in the group exposed to filtered air (p < 0.01). Increases in P(enh) were dependent on the ozone concentration. Although the protein levels of eNOS and iNOS determined were within normal levels, the amount of nNOS protein was markedly elevated in airway tissue homogenates of the group exposed to 2-ppm ozone. CONCLUSION: These findings demonstrate that the nNOS isoform may be involved in airway obstruction in mice exposed to ozone. PMID- 12584399 TI - Non-responding 'tension pneumothorax' following stab wounds. PMID- 12584400 TI - Interstitial lung disease in an intravenous drug user. PMID- 12584401 TI - A case of acute pneumonitis induced by injection of silicone for colpoplasty. AB - Silicone fluid (polymethylsiloxane) is a liquid polymer that had been widely used in breast augmentation and other cosmetic procedures. However, many adverse reactions after silicone injections into human beings have been reported including adverse pulmonary sequelae. We report a case of a 46-year-old woman who suffered from acute pneumonitis that was induced by illicit injection of silicone into the vaginal wall for colpoplasty. The pulmonary involvement was documented by transbronchial lung biopsy and bronchoalveolar lavage where examination with the electron microscope disclosed the presence of large quantities of pleomorphic cytoplasmic inclusions in alveolar macrophages. Clinicians should be aware of this unusual complication of injection of silicone. PMID- 12584402 TI - Obstructive sleep apnea syndrome as first manifestation of pharyngeal non Hodgkin's lymphoma. AB - Obstructive sleep apnea syndrome (OSAS) is characterized by repetitive episodes of upper airway obstruction during sleep that provoke an abnormal number of apneas and hypopneas, leading to arousals and, as a result, to an altered sleep architecture. Here we present a patient with clinical symptoms characteristic of OSAS. During follow-up, pharyngeal non-Hodgkin's lymphoma was diagnosed and treated, with a nearly complete normalization of polysomnography. PMID- 12584403 TI - Primary endobronchial actinomycosis associated with broncholithiasis. AB - Primary endobronchial actinomycosis is an extremely rare disease that presents with endobronchial mass. We report 2 cases of primary endobronchial actinomycosis associated with broncholithiasis. There was no foreign body material, suggesting that these broncholiths were formed endogenously. Even though it is very rare, endobronchial actinomycosis should be included in the differential diagnosis of calcified endobronchial mass, especially when there is no clinical or radiological evidence of a granulomatous infection, such as tuberculosis, and the yellowish materials obstructing the bronchi are seen during bronchoscopy. PMID- 12584404 TI - Pleural effusion with splenic rupture as manifestations of recurrence of sarcoidosis following prolonged remission. AB - A 55-year-old man presented with a 3-week history of dry cough and left pleuritic chest pain with a new exudative pleural effusion. Sixteen years earlier, he was diagnosed with sarcoidosis presenting with hilar lymphadenopathy, erythema nodosum, mildly disturbed liver function tests and noncaseating granulomata on liver biopsy, with no evidence of pulmonary parenchymal disease. He was treated with prednisone and in recent years maintained at a low daily dose, until it was eventually discontinued two years prior to his present illness. There was no evidence of infection or malignancy, and the fluid resolved following treatment with naproxen. Three weeks later the patient presented with sudden onset of dyspnea and left chest pain. After starting intravenous heparin for suspected pulmonary emboli, the patient developed hemodynamic instability which was accompanied by abdominal tenderness and decreasing hematocrit. Splenic rupture was diagnosed, and the patient underwent splenectomy. Pathology specimens revealed a hemorrhagic infarct with subcapsular hematoma, and numerous noncaseating granulomata within the splenic tissue. This patient had recurrent sarcoidosis with two rare manifestations of the disease, 2 years after withdrawing low dose prednisone, given for a prolonged time. The possibility of reemergence of the disease in organs other than the organs involved in the initial presentation should always be considered in sarcoidosis. PMID- 12584405 TI - Pleural effusion due to Histoplasma capsulatum and idiopathic CD4 lymphocytopenia. AB - Acute histoplasmosis is usually a benign, self-limited infection in endemic areas. Since protection against Histoplasma capsulatum infection requires specific, cell-mediated immunity, histoplasmosis is well documented in patients with acquired T cell deficiencies e.g. due to HIV infection. We report here for the first time a case of pleural effusion due to H. capsulatum infection in a patient with idiopathic CD4 lymphocytopenia (ICL). A 25-year-old woman presented with chest pain, dyspnea on exertion and a moderate weight loss. Chest X-ray showed a large left pleural effusion, and thoracentesis yielded an exudate. Histologic examination of pleural biopsies identified H. capsulatum. Laboratory tests revealed lymphocytopenia with low CD4+ T cell counts (<100/microl) and a decreased CD4/CD8 ratio. Serology, including HIV, was repeatedly negative. The diagnosis of pleural effusion due to H. capsulatum infection in a patient with idiophatic ICL was established. There was no evidence of any other opportunistic infection. Treatment with itraconazole was initiated and pleural effusion resolved within 2 weeks of treatment. Moreover, the patient was found to have idiopathic thrombocytopenic purpura, as confirmed by the detection of autoantibodies against thrombocytes. In a 1-year follow-up, the patient remained asymptomatic without relapse or any new infection. Treatment with itraconazole was given for 12 months. Because of persistent CD4+ T cell counts below 100/microl, prophylactic antibiotic treatment is continued. PMID- 12584406 TI - Neuroimmunomodulation and immune privilege: the role of neuropeptides in ocular immunosuppression. AB - Regional immunoregulatory mechanisms insure that the most effective immune defense mounted is in proportion with preserving unique tissue functionalities. Immune-privileged tissues, such as the eye, are tissue sites of extreme regional immunoregulation. They have evolutionarily adapted several mechanisms to prevent the induction of inflammation within their tissue microenvironment. With over half a century of experimental examinations of ocular immune privilege, only recently have we come to understand that neuropeptides constitutively present in ocular tissues are part of the mechanisms of immune privilege. PMID- 12584407 TI - Role and mechanisms of interleukin-1 in the modulation of neurotoxicity. AB - OBJECTIVE: Recent studies on cerebral ischemia in the rat have demonstrated that administration of interleukin-1 receptor antagonist (IL-1ra) markedly reduces the volumes of infarcts which are associated with N-methyl-D-aspartate (NMDA) mediated neurotoxicity. These observations suggested that endogenous interleukin 1 (IL-1) may be involved in the mediation of excitotoxic neuronal injury following ischemia. METHOD: In the present studies, we examined the role of interleukin-1beta (IL-1beta) in NMDA-related and microglia-induced excitotoxicity in cocultures of mixed neurons and microglia. RESULTS: Our observations in these mixed cultures indicated that addition of IL-1beta exaggerated NMDA and glutamate evoked hippocampal neuron death. Addition of microglia, activated by lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma), to cocultures of cortical neurons and glia induced significantly greater neurotoxicity when compared with cocultures of cortical neurons and untreated microglia. This neurotoxicity did not require that activated glia be in cell-to-cell contact with neurons. Addition of either IL-1ra or the NMDA receptor antagonist MK-801 to cocultures of cortical neurons and activated glia partially reversed the neuronal damage mediated by activated microglia. Finally, IL-1beta concentrations in the supernatant of cocultures of cortical neurons and microglia treated by LPS and IFN-gamma were markedly increased when compared with coculture of neurons with untreated microglia. CONCLUSION: These results suggest that both the NMDA receptor and the IL-1 receptor are involved in microglia-mediated neurotoxicity. PMID- 12584408 TI - Anti-inflammatory effects of alpha-melanocyte-stimulating hormone in celiac intestinal mucosa. AB - OBJECTIVES: The peptide alpha-melanocyte-stimulating hormone (alpha-MSH) possesses potent anti-inflammatory activities and has been previously implicated in the endogenous control of inflammatory reactions. The aim of the present research was to determine whether alpha-MSH and its receptors participate in a localized anti-inflammatory response in the duodenal mucosa of celiac patients. METHODS: Three series of experiments were performed, using duodenal biopsy pairs from 53 adult celiac patients and 14 normal subjects, in order to determine: (1). mucosal immunoreactivity for alpha-MSH and melanocortin receptors (MCRs), and gene expression of alpha-MSH precursor pro-opiomelanocortin and MCRs; (2). alpha MSH and inflammatory cytokine production by duodenal specimens in vitro, and the influence of synthetic alpha-MSH on such cytokine production, and (3). the influence of stimulation with gliadin (the subfraction of gluten that is toxic to patients with celiac disease) on alpha-MSH and cytokine production in vitro and the effect of alpha-MSH on gliadin-stimulated cytokine production. RESULTS: Elements of a localized anti-inflammatory influence based on alpha-MSH and its receptors were found: duodenal mucosa showed immunostaining for alpha-MSH and two of its receptor subtypes, MC1R and MC5R. alpha-MSH and MC1R immunoreactivity was more intense in specimens from celiac patients. Release of interleukin 6 from gliadin-stimulated duodenal mucosa was inhibited by synthetic alpha-MSH in vitro. CONCLUSIONS: Presence of alpha-MSH and its receptors in celiac mucosa suggests the presence of a local reaction to control the inflammatory response elicited by gliadin. In selected cases of refractory celiac disease, treatment with exogenous peptides might be considered. PMID- 12584410 TI - Psychological stress increases human T cell apoptosis in vitro. AB - OBJECTIVES: Recent studies have shown that apoptosis is involved in stress responses. The present study examined if stressors increase in vitro apoptosis of peripheral blood T lymphocytes in a dose-dependent manner. METHODS: Daily subjective stress was quantitatively analyzed in 40 nonsmoking men with a daily hassles questionnaire. Apoptosis of T lymphocytes was measured by flowcytometry using Annexin V/PI double staining method after 0, 12, and 24 h of culture in the presence or absence of dexamethasone (DEX). Using a cross-sectional design, the current study examined the relationship between stress and in vitro apoptosis of T cells. RESULTS: Results showed that apoptosis of T lymphocytes in vitro has a significant correlation with stress and age. Stress was positively correlated with percentage of apoptosis in T cells after 12 h of culture, irrespective of DEX treatment. Age was positively correlated with the percentage of T cell apoptosis after 0 and 12 h of coculture with DEX. CONCLUSIONS: These results indicate that age-related apoptosis and stress-related apoptosis of T cells are modulated through different mechanisms. This is the first study to show that in vitro lymphocyte apoptosis is influenced by daily stress in a dose-dependent manner. PMID- 12584409 TI - Neuropeptide-induced inhibition of IL-16 release from eosinophils. AB - OBJECTIVES: Eosinophils are prominent inflammatory cells that respond to peripheral neuropeptides in vitro and in vivo. At inflammatory sites, cytokines activate distinct cellular and biochemical pathways that act in a coordinated fashion. We investigated whether peripheral neuropeptides can act as immunomodulators by influencing cytokine release from eosinophils. METHODS: Human eosinophils were enriched using magnetic cell sorting, and IL-16 levels in supernatants from maintained eosinophils were measured by ELISA. Biological activity of IL-16 was confirmed in lymphocyte chemotaxis assays. RESULTS: Substance P, vasoactive intestinal polypeptide, calcitonin gene-related peptide and secretoneurin reduced the IL-16 release from eosinophils; this effect was additive to that observed with GM-CSF or IL-5. Decreased IL-16 levels in supernatants resulted in reduced lymphocyte chemotaxis, whereas supernatants derived from SP-treated eosinophils stimulated lymphocyte chemotaxis, even though IL-16 was decreased. CONCLUSIONS: Results suggest that distinct neuropeptides are able to reduce the number of lymphocytes at inflammatory sites during existing eosinophilia by inhibiting eosinophil IL-16 release, thus attenuating the pro inflammatory action of lymphocytes and monocytes. PMID- 12584411 TI - Expression of substance P and its neurokinin-1 receptor on thymocytes: functional relevance in the regulation of thymocyte apoptosis and proliferation. AB - Herein, we provide evidence of the expression and function of substance P (SP) and neurokinin-1 receptor (NK-1R) in the rat thymus. In situ hybridization evidenced NK-1R mRNA mainly in the thymic medulla, and Northern blot analysis of mRNA from FACS-sorted thymocytes identified NK-1R on CD4+, CD8+ and double positive subpopulations. With flow cytometry, it could be seen that NK-1R was expressed on the majority of CD5+ thymocytes, and it was identified by Western blot analysis as two bands migrating at 44 and 54 kD. SP administration rescues thymocytes from spontaneous and NK-1R antagonist (SR140333)-induced apoptosis and stimulates concanavalin A (ConA)-induced thymocyte proliferation, CD25 expression and IL-2 production, whereas SR140333 exerts inhibitory effects on these functions. We also demonstrated the expression of mRNA for the SP precursor preprotachykinin-A in the thymic medulla and purified CD5+ thymocytes. SP protein was detected on 40% of CD5+ thymocytes and identified as a band of 1.3 kD by Western blot analysis. Finally, thymocytes spontaneously released SP, which was increased upon ConA or CD3 stimulation. PMID- 12584412 TI - Transient global amnesia: a clinical and sonographic study. AB - BACKGROUND: The aetiology of transient global amnesia (TGA) is still unknown. The aim of this study was to identify potential risk factors for TGA, vascular risk factors, the role of patent foramen ovale (PFO) and of retrograde jugular venous flow. METHODS: 138 subjects entered the study, including 48 patients with TGA, 42 age-matched patients with transient ischaemic attack (TIA) and 48 controls. PFO was studied by contrast transcranial duplex sonography. Retrograde jugular venous flow was tested with air contrast ultrasound venography (ACUV). RESULTS: TGA patients and controls showed a lower prevalence for vascular risk factors than TIA patients. No statistical difference was found between the 3 groups with regard to PFO. ACUV detected jugular valve incompetence in 72.9% TGA, 35.7% TIA and 39.5% controls (TGA vs. TIA and TGA vs. controls p < 0.01). CONCLUSIONS: TGA patients have fewer vascular risk factors than TIA patients. Paradoxical embolism due to PFO as a cause of TGA is not confirmed in our study. Cerebral venous hypertension due to incompetence of the internal jugular valve may play a role in the pathogenesis of TGA. PMID- 12584413 TI - Progressive nonfluent aphasia and subsequent aphasic dementia associated with atypical progressive supranuclear palsy pathology. AB - We describe a right-handed man who developed progressive nonfluent aphasia and apraxia of speech beginning at age 71. By age 74 he had behavioral changes, aphasic dementia, as well as mild parkinsonism; extraocular movements were normal except for mild limitation of upgaze. Serial neuropsychometric testing and single photon emission computed tomography (SPECT) scans showed progressive changes reflecting left>right cerebral hemisphere dysfunction. Neuropathologic examination revealed findings characteristic of progressive supranuclear palsy (PSP) except that the cortical pathology was more widespread than is typical of PSP. We conclude that the clinical manifestations in this case were more similar to the syndrome of progressive nonfluent aphasia with subsequent aphasic dementia and mild parkinsonism, rather than those of PSP. Hence, PSP can present clinically as an atypical dementing syndrome dominated by progressive aphasia/apraxia of speech. PMID- 12584414 TI - Early prediction of neurological outcome after cardiopulmonary resuscitation: a multimodal approach combining neurobiochemical and electrophysiological investigations may provide high prognostic certainty in patients after cardiac arrest. AB - A reliable and reproducible method for precisely predicting the neurological outcome of patients with hypoxic-ischemic encephalopathy after cardiac arrest is urgently needed in neurological intensive care units. We prospectively investigated the predictive power of serum concentrations of neuron-specific enolase (NSE) and protein S-100B (S-100B) measured on days 1, 2, 3 and 7 as well as somatosensory-evoked potentials (SEPs) recorded within 48 h and on day 7 after cardiopulmonary resuscitation (CPR) in 27 patients (14 females, 13 males; mean age 61.3 +/- 17.3 years) with hypoxic-ischemic encephalopathy. During the first 7 days after CPR, median values of NSE and S-100B were increased in patients who remained unconscious after CPR compared to those patients who regained consciousness (significance up to < or =0.001). The best predictor of negative outcome was an NSE cutoff point > or =43 microg/l on day 2; this had a sensitivity of 90.9% and a specificity of 100%. Additional use of S-100B on day 2 did not increase sensitivity, but this could be markedly increased by combining NSE and S-100B on days 1, 3 and 7. SEPs showing bilateral loss of cortical responses identified patients who did not regain consciousness with a specificity of 100%. PMID- 12584415 TI - Mutation screening and association analysis of the parkin gene in Parkinson's disease patients from South-West China. AB - Various deletions and point mutations in the Parkin gene have been strongly associated with Parkinson's disease (PD) and parkinsonism, especially when the onset occurs at a young age. In this study, we screened 25 "early-onset" (<49 years at onset) and 91 later-onset PD patients from a Han Chinese population from South-West China for deletions and mutations in the Parkin gene. We found no deletions or point mutations in exons 1-12 of the Parkin gene using direct sequence analysis and only detected the common Ser167Asn polymorphism. We analysed Ser167Asn in 116 patients with sporadic PD and 124 controls, matched for age and gender. There were significant differences in allele and genotype frequency between PD patients, with the 167Asn allele more common in cases than controls (46.6 vs. 35.1%; chi(2) = 6.54, p = 0.011, odds ratio = 1.61, 95% confidence interval, CI, 1.10-2.37), as was the 167Asn genotype (17.3 vs. 11.3%; p = 0.04). The frequency of the 167Ser genotype was significantly lower in PD patients than in controls when compared with that of the other two genotypes combined (chi(2) = 7.84, p = 0.005, odds ratio = 0.46, 95% CI 0.25 - 0.82). No significant differences in the frequencies of the allele and genotypes were found between patients classified into two groups according to symptoms at onset (chi(2) = 0.191, p = 0.66, odds ratio = 1.12, 95% CI 0.65-1.95; chi(2) = 0.24, p = 0.887) or age of onset (p = 0.787). In summary, homozygous deletion mutations and point mutations in exons 1-12 of the Parkin gene were not detected in this Han Chinese population, although we cannot exclude compound heterozygous deletions. In addition, our study suggests that the variant 167Asn increases the risk of developing PD. PMID- 12584416 TI - Survival of cardiac function after brain death in patients in Kuwait. AB - BACKGROUND: Persistent cessation of all cerebral and brainstem function (brain death) is accepted in most countries as legal evidence of death. It is presumed that cardiac function will cease within a short time after brain death has occurred. In some countries, such as Kuwait, tradition and practice discourage application of the brain death criteria despite legal acceptance. OBJECTIVE: The study was designed to assess the duration of persistence of cardiac function in patients after the diagnosis of brain death had been made on the basis of generally accepted criteria. METHODS: We evaluated how long cardiac function persisted after brain function had ceased in 40 patients in Kuwait who were admitted to hospital and died during the 10-year period 1992-2001. RESULTS: It was found that the mean persistence of cardiac function after brain death was 8.20 days and the median survival time was 6 days. Two thirds of the patients survived longer than a week, but none had cardiac function for longer than 30 days. CONCLUSION: The study confirms that brain death is not automatically followed immediately by cessation of all other body functions. It may be speculated therefore that whole-body homeostasis is not as intimately associated with brain function as has hitherto been thought. PMID- 12584417 TI - Associated ischemic lesions in intracranial hemorrhage: MRI study. AB - Cerebral small-vessel disease is an important cause of a hemorrhagic stroke (HS) or a certain type of ischemic stroke (IS). Using magnetic resonance imaging, the frequency and the pattern of ischemic lesions between an HS group and an IS group were compared. Sixty-eight patients with HS and 104 patients with IS as a control group were enrolled in this study. The rate of grade 2 and 3 periventricular white matter lesions (WML) in the HS group was similar to that in the IS group. However, grade 2 and 3 lacunes (LAC) were more frequent in the HS group. These results show that the incidence of concomitant silent LAC is different from that of WML in HS patients. This suggests that the pathophysiology of WML and that of LAC is different and that the pathophysiology of HS is closely associated with that of LAC. PMID- 12584418 TI - Relationship between the number of involved cranial nerves and the percentage of lesions located in the cavernous sinus. AB - The main objective of this study is to investigate the relationship between the number of cranial nerves passing through the cavernous sinus affected by a lesion and the percentage of lesions located in the cavernous sinus itself. We collected cases retrospectively and classified them into groups according to the number of involved cranial nerves. The diagnoses were confirmed either with MRI, angiography or pathological examination. Sixty-eight cases in total were collected. In the increasing order of cranial nerve involvement (from 1 to 4 nerves), 17.7, 44.4, 56.3 and 77.8% of cases were found to have lesions in the cavernous sinus. A linear regression model with a least square estimator showed an approximately linear correlation. We concluded that there was an approximately linear correlation between the number of cranial nerves affected by a lesion and the percentage of lesions located in the cavernous sinus. PMID- 12584419 TI - Collet-Sicard syndrome disclosing glomus tumor of the skull base. PMID- 12584420 TI - Stroke: trial by jury. AB - Trials of pharmacological intervention for acute stroke show several points of similarity with legal trials by jury. The relationships between sponsoring organisation and regulatory authorities and the roles of the many other participants are described. Only through a partnership can the common aim of developing improved treatment for patients with stroke be realised. Excellent structures and procedures have recently developed which minimise the risk of undertaking impractical, invalid or unreported trials. This will lead to greater confidence in our ability to identify a successful treatment and to discount worthless approaches. PMID- 12584421 TI - Acute stroke trials: the problems of local investigators? AB - During stroke trials local investigators have to face many practical problems and time consuming procedures (filling in huge case report forms, performing repeat blood sample drawings for pharmacokinetic studies etc.) which, however, simply require organizational structures which is understood to be necessary to be able to conduct such kind of studies. Other, and most worrisome problems, are indeed to be solved when a sponsored research may rise potential ethical issues, or when academic research proposals clash with the interest of pharmaceutical companies or find difficulties in being funded by public institutions. It is just a greater involvement of these latter, possibly free from bureaucratic laces, which might help a balance to be struck between academic and industrial aims. PMID- 12584422 TI - The role of the data safety and monitoring committee in stroke trials. AB - There are many good reasons why any human experiment should have appropriate oversight. In clinical trials, the data safety and monitoring committee (DSMC) serves to protect the interests of the patients. In stroke trials, its members usually together represent a broad expertise in neurology, internal medicine, cardiology and biostatistics. Independency and integrity are key words, especially in the relationship to the sponsor of the trial. The DSMC members should have no long-term financial strings to the sponsor, and they should not be personally involved in patient recruitment and evaluation. The DSMC provides recommendations to the steering committee of the trial on whether to continue or to stop patient recruitment and on protocol amendments that may be necessary to protect the safety of the patients. PMID- 12584423 TI - Acute stroke: trial by jury. The toothless tiger (role of the principal investigator). AB - All trials require a principal investigator to take the lead role and act as main contact amongst the various groups of participants. His role ranges from assistance in protocol design; through selection of investigators, monitoring of study conduct and responding to Safety Committee recommendations; to involvement in study reporting and in maintaining access to the data set after the conclusion of the trial. This article describes the role of the principal investigator and Steering Committee in more detail and discusses the issues that arise out of the organisational requirements of modern clinical trials and the potential tensions between the various participating groups. PMID- 12584424 TI - The scribe of stroke trials. AB - The responsibility for reports about drug trials in medical journals should lie with the clinicians in the steering committee, not with the industrial sponsor. Examples of undue influence of sponsors on the conduct and analysis are the choice of surrogate outcome events, changes in the protocol during the trial, discontinuation of the study, post hoc analyses and suppression of publication. Since this essay was written, the International Committee of Medical Journal Editors ("Vancouver Group") has issued the condition that authors of submitted manuscripts should guarantee their independence in data analysis and reporting. Nevertheless, this "security check" occurs at a late stage and may not apply to journals that are less well known. Institutional Review Boards should routinely take the matter of independence into account. Also, European guidelines should be rewritten to reflect these fundamental issues rather than a multitude of details. In fact not sponsors or physicians but patients and the general public are the true owners of trial data. PMID- 12584425 TI - He who pays the piper calls the tune: the role of the industry sponsor in acute stroke trials. AB - Ischemic stroke remains a complex neurobiological and clinical disease, and stroke drug development remains time consuming and costly. In addressing these challenges, the pharmaceutical industry has a principal responsibility to ensure that the acute stroke trials it sponsors are ethically conducted, scientifically sound and satisfy global regulatory requirements. The industry must also be a leader in promoting the innovative and effective partnerships between the public, private and academic sectors that are necessary if new therapies for patients with acute stroke are to be developed. PMID- 12584426 TI - Non-drug therapies for dementia: an overview of the current situation with regard to proof of effectiveness. AB - An overview will be presented of music therapy, art therapy, movement therapy and reminiscence therapy, memory training, reality orientation, validation therapy, self-maintenance therapy, behaviour therapy, milieu therapy and staff training. The overview will examine the aims of each, the principles on which procedures are based and the proof of their effectiveness. The principal aim of non-drug therapies is to influence symptomatic dementia beneficially and to improve the abilities remaining to the patient. The potential benefits are usually deduced from studies made without control groups. At the present time, proof of the effectiveness of these therapies is still lacking as controlled, randomized studies have yet to be conducted, and so a fundamental evaluation of the therapeutic benefits of non-drug therapies in the treatment of dementia cannot yet be made. PMID- 12584427 TI - Neuropsychological profile of patients with Parkinson's disease without dementia. AB - Cognitive deficits are often associated with Parkinson's disease (PD), although their prevalence in PD patients without dementia is still unknown. In order to describe the neuropsychological profile of PD patients without dementia, a sample of 103 PD patients was compared with a control group consisting of 38 healthy elderly subjects. Psychometric assessment consisted of the Mini Mental State Examination, the Dementia Rating Scale and a battery of neuropsychological tests. The Beck Depression Inventory was used to assess depression in PD patients. Dementia was diagnosed in 27 patients. Among non-demented subjects, 34 (45%) had no cognitive impairment and 42 (55%) had a mild cognitive impairment. Subjects with mild cognitive impairment were older, had a later onset of the disease, and more severe motor symptoms than cognitively intact subjects. Identification of mild cognitive impairment is important, since these symptoms are important for patient management and may also facilitate to determine prognosis. PMID- 12584428 TI - Kinematic analysis of handwriting movements in patients with Alzheimer's disease, mild cognitive impairment, depression and healthy subjects. AB - A variety of studies have demonstrated that motor disorders, parkinsonism and extrapyramidal motor symptoms (EPMS) are common in patients with Alzheimer's disease (AD). Several studies have reported an association of EPMS with severity, progression and poor prognosis of AD. The majority of these studies used clinical assessments for the rating of EPMS. In this study, kinematic handwriting analysis was used to quantify differences in fine hand motor function in patients with probable AD and mild cognitive impairment (MCI, as an assumed initial stage of AD) compared to depressed patients and healthy controls. Both patients with MCI and patients with probable AD exhibited loss of fine motor performance. Movements of AD patients were significantly less regular than those of healthy controls. PMID- 12584429 TI - Are MRI white matter lesions clinically significant in the 'old-old'? Evidence from the Sydney Older Persons Study. AB - BACKGROUND: The number of individuals aged over 80 years is the fastest increasing group in developed countries. White matter lesions (WML) observed on magnetic resonance imaging (MRI) have uncertain clinical significance, particularly in the old. OBJECTIVES: To determine the prevalence of periventricular and deep WML in survivors of an original cohort of randomly selected elderly community dwellers, and to examine their associations with clinical markers of vascular and extrapyramidal disorders of ageing, as well as quantitative cognitive measures. METHODS: Brain MRI, lifestyle interview, cognitive testing and medical examination were administered to 122 participants from the Sydney Older Persons Study 6-year review (mean age: 85.5 years). Apolipoprotein E (ApoE) genotype was also established. Presence and severity of periventricular and deep WML were ascertained using semi-quantitative rating methods and their relations to the cognitive and clinical variables investigated. RESULTS: Periventricular WML were present in all participants in similar severity for all three regions sampled. In contrast, a gradient of severity was observed for the deep WML: most severe in the parietal region, followed by the frontal and occipital regions, and least severe in the temporal region. Associations with gender or with the ApoE epsilon4 allele were non-significant. WML were inconsistently associated with age and cognitive functioning or with the clinical markers of dementia. No frontal specificity emerged. Examination of individual lesion types did not change the general pattern of associations. Supporting evidence for a threshold effect was observed on some measures. CONCLUSIONS: WML are extremely common in elderly, non-demented individuals. Unlike in younger individuals, MRI abnormalities may not be evidence of a current pathological process and their importance may change with advancing age. PMID- 12584430 TI - Interaction of the H63D mutation in the hemochromatosis gene with the apolipoprotein E epsilon 4 allele modulates age at onset of Alzheimer's disease. AB - The H63D mutation in the hemochromatosis gene (HFE) has recently been considered as a risk factor in Alzheimer's disease (AD) with advancing age at onset of the disease, independently of the apolipoprotein E (ApoE) epsilon4 allele effect. We examined the distribution of the H63D mutation and ApoE genotypes as a function of age at AD onset in 328 patients with sporadic AD. Our data show that the mutant H63D allele potentially interacts with the ApoE epsilon4 allele to significantly reduce age at onset of AD compared to ApoE epsilon4 carriers alone, but has no effect on age at onset in ApoE epsilon4 non-carriers. PMID- 12584431 TI - Homocysteine as a risk factor for cognitive impairment in stroke patients. AB - BACKGROUND: Elevated total homocysteine (tHcy) levels are associated with an increased risk of cerebrovascular disease. It is uncertain whether tHcy is also an independent risk factor for cognitive impairment. METHODS: We examined 95 stroke subjects 3 months after their strokes, and 55 healthy comparison subjects, with a detailed neuropsychological assessment, and MRI brain scans in a proportion (n = 97). Baseline measurements of tHcy, serum folate and B(12), creatinine and plasma fibrinogen levels were obtained. RESULTS: tHcy levels were higher in the stroke subjects by a mean 34%. These levels were significantly correlated with the first factor of a principal component analysis of the neuropsychological data, after controlling for age, folate, B(12) and creatinine levels. The correlation of Hcy levels was particularly significant with frontal executive functioning and attention. tHcy levels were significantly correlated with number of infarcts and total stroke volume in the stroke group, but not with T(2)-weighted deep white matter hyperintensity scores, after correction for age. In the control group, tHcy levels were significantly correlated with ventricle-to brain ratios as measures of brain atrophy. CONCLUSION: This study provides evidence that high tHcy levels are associated with cognitive impairment, in particular that of frontal-executive function. The major component of this association is accounted for by small and large strokes, but non-vascular neurotoxic effects of tHcy also appear to play a role. tHcy must receive greater attention as a risk factor for cognitive impairment. PMID- 12584432 TI - Recognition of facial expressions and emotional situations in patients with dementia of the Alzheimer and vascular types. AB - The aim of this study was to investigate the ability of patients with dementia to recognize facial expressions and emotional situations. We evaluated 16 patients with dementia of the Alzheimer type (DAT) and 15 with vascular dementia (VD) for general cognition, discrimination of facial expressions and individual faces, and recognition of facial expressions and emotional situations. VD patients performed significantly worse than DAT patients at recognizing facial emotions, even though there was no difference between them in their general cognition and visuoperceptual abilities. There was no significant difference between them in their ability to recognize emotional situations. The results of this and past studies suggest that caregivers in nursing homes and hospitals need to be aware that VD patients lose the ability to comprehend facial expressions. PMID- 12584433 TI - Cerebrospinal fluid beta-amyloid 42 is reduced before the onset of sporadic dementia: a population-based study in 85-year-olds. AB - Deposition of beta-amyloid (Abeta) is an early pathogenic event in Alzheimer's disease (AD). We measured Abeta42 and Abeta40 in cerebrospinal fluid (CSF) in a population-based sample of 85-year-olds, 27 demented and 35 non-demented. During the following 3 years, 7 of the 35 non-demented individuals had developed dementia, while 28 remained non-demented. Reduced CSF levels of both Abeta42 (p = 0.001) and Abeta40 (p = 0.0001) were found in patients with manifest AD and vascular dementia at the age of 85. Non-demented individuals who developed dementia during follow-up had lower levels of CSF- Abeta42 (p = 0.003), but not CSF-Abeta40 (p = 0.96), than those who remained non-demented. The odds ratio for development of dementia was 8.2 (p = 0.027) for individuals in the lower 50th percentile of CSF-Abeta42, while none of those in the highest 33rd percentile of CSF-Abeta42 developed dementia during follow-up. There were no significant differences between carriers and non-carriers of the apolipoprotein E epsilon4 allele regarding CSF-Abeta42 or CSF-Abeta40. Our study suggests that low CSF Abeta42 is found also in an unselected population-based sample of old demented patients and provides the first evidence of a disturbance in the metabolism of Abeta, specifically involving Abeta42, before the onset of clinical symptoms in AD. PMID- 12584434 TI - A novel family-specific translocation t(2;20)(p24.1;q13.1) associated with recurrent abortions: molecular characterization and segregation analysis in male meiosis. AB - In the present study, we present a novel reciprocal translocation t(2;20)(p24.1;q13.1) and its segregation in a three generation family. The rate of miscarriages (50%) in pregnancies from male translocation carriers could be explained by unbalanced translocation-bearing spermatozoa found with a frequency of approximately 55% in the entire sperm population of a t(2;20)(p24.1;q13.1) carrier. These imbalanced spermatozoa mainly present as 2, der(20) and der(2), 20 missegregated (approximately 46%) while adjacent 2 and 3:1 segregation patterns account for approximately 5% and 4% of imbalances, respectively. While the translocation is associated clearly with an increased risk of early abortions (7/12) in both male and female carriers, no malformed livebirths were observed. Our results suggest complete embryonic lethality of imbalanced offspring. With respect to a high rate of segregation to 2, der(20) and to der(2), 20 imbalanced spermatozoa in male translocation carriers and with respect to known cases of partial trisomy 2p and 20q we consider that their corresponding monosomies result in fetal loss. This is the first study reporting multiple abortions associated with partial monosomy 20q13.1-->qter and 2pter-->p24.1 and the first report on the frequency of chromosomal imbalances in gametes of a male t(2;20)(p24.1;q13.1) heterozygote. PMID- 12584435 TI - Stabilization of a terminal inversion duplication of 8p by telomere capture from 18q. AB - Terminal inversion duplications of the short arm of chromosome 8 are one of the more common chromosome rearrangements in humans. We report an infant with multiple congenital anomalies, in whom karyotype analysis showed a terminal inversion duplication of 8p including additional material at the distal end of the derivative chromosome, shown to be of chromosome 18q origin. Terminal inversion duplications of 8p are the result of meiotic recombination between inverted olfactory gene receptor repeats in 8p. This recombination generates a dicentric intermediate that breaks during anaphase, and the broken chromosome end is stabilized by telomere healing or telomere capture. The origin of the telomeric region in the majority of constitutional chromosome deletions studied to date was shown to be from telomere healing; the de novo addition of telomeric repeats. In the proband a cytogenetically detectable piece of chromosome 18q was present on the distal end of the derivative 8, suggesting that this chromosome was stabilized by telomere capture of 18q. FISH analyses of additional cases may yield information as to whether telomere capture or telomere-healing events are the predominant mechanism of chromosome stabilization in terminal inversion duplications of 8p. PMID- 12584436 TI - Prolonged culture of normal chorionic villus cells yields ICF syndrome-like chromatin decondensation and rearrangements. AB - Untreated cultures from normal chorionic villus (CV) or amniotic fluid-derived (AF) samples displayed dramatic cell passage-dependent increases in aberrations in the juxtacentromeric heterochromatin of chromosomes 1 or 16 (1qh or 16qh). They showed negligible levels of chromosomal aberrations in primary culture and no other consistent chromosomal abnormality at any passage. By passage 8 or 9, 82 +/- 7% of the CV metaphases from all eight studied samples exhibited 1qh or 16qh decondensation and 25 +/- 16% had rearrangements in these regions. All six analyzed late-passage AF cultures displayed this regional decondensation and recombination in 54 +/- 16 and 3 +/- 3% of the metaphases, respectively. Late passage skin fibroblasts did not show these aberrations. The chromosomal anomalies resembled those diagnostic for the ICF syndrome (immunodeficiency, centromeric region instability, and facial anomalies). ICF patients have constitutive hypomethylation at satellite 2 DNA (Sat2) in 1qh and 16qh, generally as the result of mutations in the DNA methyltransferase gene DNMT3B. At early and late passages, CV DNA was hypomethylated and AF DNA was hypermethylated both globally and at Sat2. DNMT1, DNMT3A, or DNMT3B RNA levels did not differ significantly between CV and AF cultures or late and early passages. The high degree of methylation of Sat2 in late-passage AF cells indicates that hypomethylation of this repeat is not necessary for 1qh decondensation. Sat2 hypomethylation may nonetheless favor 1qh and 16qh anomalies because CV cultures, with their Sat2 hypomethylation, displayed 1qh and 16qh decondensation and rearrangements at significantly lower passage numbers than did AF cultures. Also, CV cultures had much higher ratios of ICF-like rearrangements to heterochromatin decondensation in chromosomes 1 and 16. These cultures may serve as models to help elucidate the biological consequences of cancer-associated satellite DNA hypomethylation. PMID- 12584437 TI - Spectral karyotype analysis of colon cancer cell lines of the tumor suppressor and mutator pathway. AB - BACKGROUND AND AIMS: Microsatellite instability (MSI) is characterized by the size variation of microsatellites in tumor DNA as compared to matching normal DNA due to defects in the mismatch repair system. To examine the chromosomal differences in microsatellite-stable (MSS) and -unstable (MSI) tumors in detail, we analyzed MSS (Caco-2, Colo-205, SW948) and MSI (HCT-15, HCT-116, LoVo) cell lines by spectral karyotyping (SKY). METHODS: SKY is a sensitive method to detect chromosome aberrations by visualizing each chromosome in a different color. Metaphases were hybridized with a SKY probe mixture. Images were visualized with the SpectraCube system and analyzed with the SKYview imaging software. RESULTS: The average number of chromosomes was 49 in LoVo, 45 in HCT-116, 46 in HCT-15, 71 in Colo-205, 89 in Caco-2 and 66 in SW-948. Three aberrant chromosomes were detected in LoVo, three in HCT-116, two in HCT-15, seventeen in Colo-205, fourteen in Caco-2 and nine in SW948. CONCLUSION: The karyotypes of MSS colon cancer cells displayed complex numerical and structural aberrations. In contrast the chromosomes of MSI colon cancer cells were mostly unaltered but displayed a few isolated numerical and structural aberrations. We speculate that these isolated aberrations may be specifically involved in the pathogenesis of MSI tumors. PMID- 12584438 TI - Physical map of the chromosome 6q22 region containing the oculodentodigital dysplasia locus: analysis of thirteen candidate genes and identification of novel ESTs and DNA polymorphisms. AB - Oculodentodigital dysplasia (ODDD) is an autosomal dominant condition with congenital anomalies of the craniofacial and limb regions and neurodegeneration. Genetic anticipation for the dysmorphic and neurologic features has been inferred in a few families. Our previous linkage studies have refined the ODDD candidate region to chromosome 6q22-->q23. In an attempt to clone the ODDD gene, we created a yeast artificial chromosome contig with 31 redundant clones spanning the region and identified and ordered candidate genes and markers. Fluorescent IN SITU hybridization mapped two of these YAC clones to chromosome 6q22.2 telomeric to a known 6q21 fragile site, excluding it as a possible cause of the suggested anticipation. We performed mutation analysis on thirteen candidate genes - GRIK2, HDAC2, COL10A1, PTD013, KPNA5, PIST, ROS1, BRD7, PLN, HSF2, PKIB, FABP7, and HEY2. Although no mutations were found, we identified 44 polymorphisms, including 28 single nucleotide polymorphisms. Direct cDNA selection was performed and fifty five clones were found to contain sequences that were not previously reported as known genes or ESTs. These clones and polymorphisms will assist in the further characterization of this region and identification of disease genes. PMID- 12584439 TI - Identification of the human ortholog of the t-complex-encoded protein TCTE3 and evaluation as a candidate gene for primary ciliary dyskinesia. AB - Primary ciliary dyskinesia (PCD) is a heterogeneous autosomal recessive disease that is caused by impaired ciliary and flagellar functions. About 50% of PCD patients show situs inversus, denoted as Kartagener syndrome. In most cases, axonemal defects in cilia and sperm tails can be demonstrated by electron microscopy, i.e. PCD patients often lack inner and/or outer dynein arms in their sperm tails and cilia, supporting the hypothesis that mutations in dynein genes may cause PCD. In order to identify novel PCD genes we have isolated the human ortholog of the murine TCTE3 gene. The human TCTE3 gene encodes a dynein light chain and shares high similarity to dynein light chains of other species. The TCTE3 gene is expressed in tissues containing cilia or flagella, it is composed of four exons and located on chromosome 6q25-->q27. To elucidate the role of TCTE3 as a candidate gene for PCD a mutational analysis of thirty-six PCD patients was performed. We detected five polymorphisms in the coding sequence and in the 5' UTR of the TCTE3 gene. In one patient a heterozygous nucleotide exchange was identified resulting in an arginine to isoleucine substitution at the amino acid level. However, this exchange was also detected in one control DNA. Our results indicate that mutations in the TCTE3 gene are not a main cause of primary ciliary dyskinesia. PMID- 12584440 TI - Identification and characterization of human taste receptor genes belonging to the TAS2R family. AB - The sense of taste is a chemosensory system responsible for basic food appraisal. Humans distinguish between five primary tastes: bitter, sweet, sour, salty and umami. The molecular events in the perception of bitter taste are believed to start with the binding of specific water-soluble molecules to G-protein-coupled receptors encoded by the TAS2R/T2R family of taste receptor genes. TAS2R receptors are expressed at the surface of taste receptor cells and are coupled to G proteins and second messenger pathways. We have identified, cloned and characterized 11 new bitter taste receptor genes and four new pseudogenes that belong to the human TAS2R family. Their encoded proteins have between 298 and 333 amino acids and share between 23 and 86% identity with other human TAS2R proteins. Screening of a mono-chromosomal somatic cell hybrid panel to assign the identified bitter taste receptor genes to human chromosomes demonstrated that they are located in chromosomes 7 and 12. Including the 15 sequences identified, the human TAS2R family is composed of 28 full-length genes and 16 pseudogenes. Phylogenetic analyses suggest a classification of the TAS2R genes in five groups that may reflect a specialization in the detection of specific types of bitter chemicals. PMID- 12584441 TI - Cytogenetic analysis of California condor (Gymnogyps californianus) chromosomes: comparison with chicken (Gallus gallus) macrochromosomes. AB - The California condor is the largest flying bird in North America and belongs to a group of New World vultures. Recovering from a near fatal population decline, and currently with only 197 extant individuals, the species remains listed as endangered. Very little genetic information exists for this species, although sexing methods employing chromosome analysis or W-chromosome specific amplification is routinely applied for the management of this monomorphic species. Keeping in mind that genetic conditions like chondrodystrophy have been identified, preliminary steps were undertaken in this study to understand the genome organization of the condor. This included an extensive cytogenetic analysis that provided (i) a chromosome number of 80 (with a likelihood of an extra pair of microchromosomes), and (ii) information on the centromeres, telomeres and nucleolus organizer regions. Further, a comparison between condor and chicken macrochromosomes was obtained by using individual chicken chromosome specific paints 1-9 and Z and W on condor metaphase spreads. Except for chromosomes 4 and Z, each of the chicken (GGA) macrochromosomes painted a single condor (GCA) macrochromosome. GGA4 paint detected complete homology with two condor chromosomes, viz., GCA4 and GCA9 providing additional proof that the latter are ancestral chromosomes in the birds. The chicken Z chromosome showed correspondence with both Z and W in the condor. The homology suggests that the condor sex chromosomes have not completely differentiated during evolution, which is unlike the majority of the non-ratites studied up till now. Overall, the study provides detailed cytogenetic and basic comparative information on condor chromosomes. These findings significantly advance the effort to study the chondrodystrophy that is responsible for over ten percent mortality in the condor. PMID- 12584442 TI - Characterization of the region containing the jcpk PKD gene on mouse Chromosome 10. AB - The jcpk gene on mouse Chromosome 10 causes a severe, early onset form of polycystic kidney disease (PKD) when inherited in an autosomal recessive manner. In order to positionally clone this gene, high resolution genetic and radiation hybrid maps were generated along with a detailed physical map of the approximately 500-kb region containing the jcpk gene. Additionally, sixty-nine kidney-specific ESTs were evaluated as candidates for jcpk and subsequently localized throughout the mouse genome by radiation hybrid mapping analysis. Previous studies indicating non-complementation of the jcpk mutation and 67Gso, a new PKD translocation mutant had suggested that 67Gso represents a new allele of jcpk. Fluorescence in situ hybridization (FISH) analysis using key bacterial artificial chromosome clones from the jcpk critical region, refined the 67Gso breakpoint and provided support for the allelism of jcpk and 67Gso. PMID- 12584443 TI - Construction of a dense comparative map between human chromosome 1p36-->p35 and swine chromosome 6 by using human sequence-tagged sites. AB - Construction of a comprehensive comparative map between swine and human chromosomes is a prerequisite, in order to select candidate swine genes for traits from the human genome database as well as to understand the evolutionary process of the two species. The present study attempted to use 910 sequence tagged sites (STSs) localized in human chromosome (HSA) 1p36-->p35 (35 Mbp) for radiation hybrid (RH) mapping to swine chromosomes (SSCs). Out of the 910 STSs subjected to amplification of swine orthologues, primer pairs for 13 STSs were found to amplify the respective orthologues and the STSs were assigned to SSCs. Eleven STSs were assigned to SSC6 in the same order as that in HSA1: SSC6cen (SHGC-150)-(A006H31)-(X82877)-(A007E03)-(IB404)-(stGDB:371372)-(stSG31658) (A009Q18)-(stSG14201/A009C01)-(H08335)-qter. One of the remaining two STSs, WI 20819, was assigned to SSCX, and the other, R91D18R, was not linked to any first generation markers of the IMpRH map with a lod score greater than 3. PMID- 12584444 TI - Localization, cDNA sequence and genomic organization of the rat seipin gene (Bscl2) and sequence analysis in inbred rat models of Type 2 diabetes mellitus. AB - Mutations in the gene encoding seipin cause Berardinelli-Seip congenital lipodystrophy 2, with symptoms including near-absence of adipose tissue and altered glucose tolerance. Radiation hybrid analysis localized the seipin gene (Bscl2) in rat to a major quantitative trait locus in rat chromosome 1 linked to glucose intolerance in the Goto-Kakizaki (GK) rat model of Type 2 diabetes. We determined the genomic organization of Bscl2 and screened coding exons and flanking intron sequences for mutations in GK, Wistar and Brown Norway rats, as well as in the Otsuka Long-Evans Tokushima Fatty (OLETF) diabetic rat. Two silent single nucleotide polymorphisms that were identified also were found in non diabetic rat strains. We conclude that mutations in the gene for seipin are unlikely to contribute to diabetes in GK and OLETF rats. PMID- 12584445 TI - "Ag-NORs" are not always true NORs: new evidence in mammals. AB - In spite of uncertainty about the biochemical processes involved, silver staining is a widely used technique for assessing the locations of active NORs in eukaryotic genomes in general, and in mammalian genomes in particular. However, following a previous study of hedgehog chromosomes, we present here a second example from two gerbil species (Rodentia, Muridae), which have several clear Ag positive signals that do not correspond to 28S rDNA clusters. Although this pattern may be characteristic of particular genomes displaying unusual heterochromatic features, our study casts doubt upon the reliability and universality of Ag-staining for detecting active NORs. PMID- 12584446 TI - Dynamics of 5S rDNA in the tilapia (Oreochromis niloticus) genome: repeat units, inverted sequences, pseudogenes and chromosome loci. AB - In higher eukaryotes, the 5S ribosomal DNA (5S rDNA) is organized in tandem arrays with repeat units composed of a coding region and a non-transcribed spacer sequence (NTS). These tandem arrays can be found on either one or more chromosome pairs. 5S rDNA copies from the tilapia fish, Oreochromis niloticus, were cloned and the nucleotide sequences of the coding region and of the non-transcribed spacer were determined. Moreover, the genomic organization of the 5S rDNA tandem repeats was investigated by fluorescence IN SITU hybridization (FISH) and Southern blot hybridization. Two 5S rDNA classes, one consisting of 1.4-kb repeats and another one with 0.5-kb repeats were identified and designated 5S rDNA type I and type II, respectively. An inverted 5S rRNA gene and a 5S rRNA putative pseudogene were also identified inside the tandem repeats of 5S rDNA type I. FISH permitted the visualization of the 5S rRNA genes at three chromosome loci, one of them consisting of arrays of the 5S rDNA type I, and the two others corresponding to arrays of the 5S rDNA type II. The two classes of the 5S rDNA, the presence of pseudogenes, and the inverted genes observed in the O. niloticus genome might be a consequence of the intense dynamics of the evolution of these tandem repeat elements. PMID- 12584447 TI - A radiation hybrid map of the X-chromosome of the dog (Canis familiaris). AB - The dog serves as an animal model for several human diseases including X chromosome diseases. Although the canine X-chromosome is one of the largest chromosomes in the dog, only a few markers have been mapped to it to date. Using a commercially available canine whole genome radiation hybrid (RH) panel we have localized 14 microsatellite markers, 18 genes and 13 STSs on the canine X chromosome, extending the total number of mapped markers to 45 covering an estimated 830 cR. Out of these 45 markers, seven distinct groups of markers could be established with an average spacing of 18.8 cR(3000) and ten markers remained unlinked. Using FISH analysis, six markers could be mapped physically to the p- or q-arm of the X-chromosome. Combined with the FISH mapping, three RH groups could be assigned to the p-arm and two RH groups to the q-arm. Comparison with the human X-chromosome map revealed conserved synteny up to 234 cR (TIMP1-ALAS2 AR-IL2RG-XIST). We show here that the similarity of the canine and human X chromosomes is the largest for any mammalian species beyond the primates. PMID- 12584448 TI - The evolutionary conservation of the mammalian peroxidase genes. AB - Myeloperoxidase (MPO), eosinophil peroxidase (EPX) and lactoperoxidase (LPO) are mammalian peroxidase enzymes possessing similar structures and functions. Here, we demonstrate that the genes encoding these molecules form a cluster on mouse chromosome 11. Genomic sequence analysis revealed that the mouse LPO gene has similar genomic organization to the corresponding human gene. Our data strongly suggest the evolutionary conservation of mammalian peroxidase genes. PMID- 12584450 TI - Barley chromosome arms longer than half of the spindle axis interfere with nuclear divisions. AB - We have tested the influence of recombinantly-elongated chromosome arms on nuclear divisions in barley and confirmed a rule according to which half the length of the average spindle axis defines the upper tolerance limit for chromosome arm length. A slightly longer chromosome arm caused incomplete separation of sister chromatids in approximately 70% of mitotic telophase cells and >2.5% of daughter cells showing a micronucleus, due to disruption of non separated sister chromatids by the newly forming cell wall. In homozygous condition, this elongated chromosome mediated a slower growth and reduced fertility of the carrier plants. Its meiotic transmission was not impaired because of the larger spindle dimensions in meiocytes as compared to those in mitotic cells. PMID- 12584449 TI - Cloning and mapping of platypus SOX2 and SOX14: insights into SOX group B evolution. AB - Group B SOX genes, the closest relatives to the sex-determining gene SRY, are thought to have evolved from a single ancestral SOX B by a series of duplications and translocations. The two SOX B genes SOX2 and SOX14 co-localize to chromosome 3q in humans. SOX2 and SOX14 homologues were cloned and characterized in the platypus, a monotreme mammal distantly related to man. The two genes were found to co-localize to chromosome 1q in this species. Proximity of the two related genes has therefore been conserved for 170 Myr, since humans and platypus diverged. The sequence similarity and conserved synteny of these group B genes provide clues to their origin. A simple model of SOX group B gene evolution is proposed. PMID- 12584451 TI - Our ethical boundaries and dilemmas. PMID- 12584452 TI - New methods of hormonal contraception. AB - Several new hormonal contraceptive methods have recently been approved by the Food and Drug Administration. most new methods combine the high efficacy of oral contraceptives with the longer-term delivery methods that eliminate the need to take a pill each day. New methods also offer reversibility and resumption of fertility if desired. This article discusses these new methods and reviews the literature on efficacy and side effects. PMID- 12584453 TI - Caring for victims of nuclear and radiological terrorism. AB - As primary care providers, we must consider the manifestations of radiation sickness and syndromes when managing radiological injuries. Assessment and treatment regimens include unique sample handling requirements to avoid self contamination and further contamination of the environment. Addressing psychological aspects and the needs of pediatric and geriatric populations is also important for effective treatment. PMID- 12584454 TI - Treatment options alleviate female urge incontinence. AB - Urge incontinence affects a considerable percent of adult women, yet remains largely untreated. It is most often a chronic condition and can be debilitating when neglected. Advanced Practice Nurses need to be aware of incontinence in their patients, determine the underlying causes and individualize treatment to each patient. Several effective treatment options exist in managing urge incontinence, including behavioral therapy, pharmacologic therapy and surgical management. The clinician initiates and follows treatment regimens, and is ideally suited to provide patients with the education and motivation required for chronic therapy. PMID- 12584455 TI - Patient education: urge incontinence. Nurse Practitioner. PMID- 12584456 TI - Advocating for NPs--go and do likewise. PMID- 12584457 TI - Effective labor relations discipline: learning the rules of the management high road. PMID- 12584458 TI - Increasing clinical, satisfaction, and financial performance through nurse-driven process improvement. AB - Providers and regulatory agencies are increasing their demands for hospitals to document clinical quality and customer satisfaction at a decreased cost. Nurse leaders search for ways to meet these requirements while retaining their most valuable resource: healthcare workers (especially nurses). First-line leadership struggles with balancing quality, cost-effectiveness, and staff retention. The author discusses the planning, methodology, implementation, and outcomes of a staff driven process improvement initiative. PMID- 12584459 TI - Patients' needs for nursing care: beyond staffing ratios. AB - The current focus on staffing ratios as a means to assure appropriate care for patients ignores the very real differences among patients in their needs for nursing care. Implementing a system that identifies these needs provides a more accurate indication of staffing requirements. In addition, storing the raw data from the system at their most basic level provides opportunities for more extensive analyses and informed, data-driven decision-making related to resource allocation, performance improvement, and productivity enhancement. PMID- 12584460 TI - Making patient-centered care come alive: achieving full integration of the patient's perspective. AB - A patient-centered model of care has profound implications for the way that care is planned, delivered, and evaluated. Although most leaders in healthcare organizations today embrace the basic tenets of a patient-centered philosophy, they often find that moving toward a patient-centered model requires an unanticipated level of commitment and significant adjustments in organizational structures. In this article, the authors describe how patients and families have been integrated into the care delivery model by involving them in planning, decision-making, and improvement processes at all levels of the organization. PMID- 12584461 TI - A model for examining predictors of outcomes of telephone nursing advice. AB - Although telephone advice nursing is the fastest-growing nursing specialty, useful information to guide managers' decisions about how best to structure and support advice services to achieve desired outcomes is unavailable. We identified issues and variables relevant to outcomes of telephone advice from the perspectives of callers, nurses, and the system. Subsequently, we derived a model for studying factors affecting nursing advice outcomes that will help managers identify modifiable factors to improve outcomes of care. PMID- 12584462 TI - The value of collaborative governance/staff empowerment. AB - OBJECTIVE: Collaborative governance is the decision-making process that places the authority, responsibility, and accountability for patient care with the practicing clinician. BACKGROUND: Collaborative governance was introduced as one of nine structures within the patient care services' professional practice model to facilitate communication and optimize staff participation in decision-making across disciplines. The concept of empowerment was used to evaluate the impact of the collaborative governance structure on members. METHODS: The purpose of the current study was to compare empowerment and power scores for both members and nonmembers of collaborative governance over a 3-year period. RESULTS: Initial survey data (baseline) from 136 staff from across disciplines was returned prior to beginning work on a committee. Study results were based on survey returns from 657 staff over a 3.5-year period. There were no significant differences between collaborative governance members and nonmembers on some demographic variables such as age, but there were significant differences on variables such as work status and education. All empowerment scores were significantly higher at the 2nd and 3rd measurement periods for collaborative governance members as compared to nonmembers. CONCLUSIONS: Findings suggest that membership on a collaborative governance committee increased staff sense of empowerment and fostered self growth and organizational development. PMID- 12584463 TI - A different age for leadership, part 1: new context, new content. AB - This is Part 1 of a 2-part article on the new and emerging characteristics and elements of leadership for changing, fast-paced organizations. As we leave the 20th century workplace and are increasingly driven by innovation and technological transformation, new roles are demanded from everyone. Leadership expression now calls for a different emphasis and skill set from those that predominated in the past century. The first article focuses on the context of leadership affecting what leaders do and how they must now behave. Part 2 will center on the activities of leadership and the new learning and skill set development that will increasingly be required of leaders in a fundamentally altered work environment. PMID- 12584464 TI - Reasons registered nurses leave or change employment status. AB - High turnover of nurses compromises patient care and adds to the cost of healthcare. To determine why nurses changed positions or left a midwestern health system, the authors surveyed registered nurses who voluntarily terminated or changed their employment status to as-needed (PRN) within a 9-month period. They discuss the most frequent reasons given for these changes and suggest solutions for retention. PMID- 12584465 TI - Errors in nursing. PMID- 12584466 TI - Genetic variability and transplantation. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize recent advances within the area of genetic polymorphisms with a specific emphasis on renal transplantation, and to discuss the potential clinical applications. RECENT FINDINGS: Due to recent advances in molecular techniques, there has been an abundance of publications describing genetic variability in molecules relevant to transplant outcome. Many studies are now demonstrating associations between polymorphisms in these candidate genes and outcomes in organ transplantation. SUMMARY: These studies emphasize the potential role of genetic variability in transplantation, and provide the rationale for large prospective studies to clearly define the potential benefits of genotyping in the risk stratification of transplant recipients. PMID- 12584467 TI - Treatment of hypertension in renal transplant recipients. AB - PURPOSE OF REVIEW: Hypertension is very common in renal transplant recipients and is a significant risk factor for mortality from cardiovascular diseases and for development of graft dysfunction. RECENT FINDINGS: Recent guidelines for the treatment of hypertension (Joint National Committee on Prevention, Detection, and Treatment of High Blood Pressure VI Report and World Health Organization Guidelines) do not directly address post-transplant hypertension. Specific recommendations for the drug treatment of hypertension in renal allograft recipients have not been given in the Clinical Practice Guidelines of the American Society of Transplantation or those of the European Renal Association. SUMMARY: The present paper summarizes some important aspects of post-transplant hypertension and discusses potential treatment strategies aimed at reducing blood pressure and thus improving patient and allograft survival. PMID- 12584468 TI - The influence of organ donor factors on early allograft function. AB - PURPOSE OF REVIEW: Postischaemic acute renal allograft failure is among the main risk factors for reduced transplant survival. Although new immunosuppressive protocols have reduced the number of acute rejections, the incidence of acute renal failure remained unchanged. On the basis of histomorphology it is not possible to predict donor kidneys at risk of subsequent failure. Some factors are associated with failure, but even combinations of these risk factors can not precisely predict the development of acute renal failure. Studies have therefore evaluated the influence of demographic donor and recipient factors on acute renal failure. New biotechnology and data mining tools are currently being used to study and identify the molecular predictors of acute renal failure. RECENT FINDINGS: Recent studies showed that donor factors contributed to approximately 40% of the variability in early allograft function. Deductive approaches identified some isolated molecular targets, such as adhesion molecules, as risk factors. Explorative analysis of the entire human genome, however, identified several predictive clusters of genes, which can be functionally grouped into categories such as cell death, stress response, cell adhesion, transcription factors, inflammatory response or cell cycle-related genes. Based on this information, preventative strategies using antisense oligonucleotides or antibodies were adopted. Clinical studies identified the use of catecholamines in the organ donor as beneficial. All these efforts aim to reduce renal tubular damage. SUMMARY: A detailed analysis of the molecular events and pathways of renal gene expression in the donor and after reperfusion, together with sophisticated data analysis tools, will provide new insights into the pathophysiology of acute renal failure. PMID- 12584469 TI - Tumours after kidney transplantation. AB - PURPOSE OF REVIEW: With the improved long-term outcome of renal allograft recipients, malignant tumours or cardiovascular disease become increasingly important. Malignant tumours develop in 15-20% of graft recipients after 10 years, and thus contribute substantially to the morbidity and mortality of these patients. In contrast to the general population, skin tumours and lymphoproliferative disorders are the most frequent malignancies in transplant recipients. Malignancies can develop in three ways: de-novo occurrence in the recipient; recurrent malignancy in the recipient; or transmission of malignancy from the donor. RECENT FINDINGS: The immunosuppressive strategies after renal transplantation differ with respect to the development of malignancies, with cell depleting antibodies being the highest risk, whereas newer immunosuppressants such as rapamycin could possess anti-tumour potential. The relationship of chronic viral infections to skin tumours and lymphoproliferative diseases has become clearer during recent years. Concomitantly, experience in the management of such diseases has grown. Furthermore, as older donors are accepted, awareness of the possibility of transferring malignancies from the donor to the recipient must increase. SUMMARY: Malignancies are a major contributor to morbidity and mortality among kidney transplant recipients as such diseases gain importance with longer graft survival. Immunosuppression and chronic viral infections in combination with the transmission of malignant cells from the donor or recurrent malignancies contribute to the increased incidence of cancer. In kidney transplant recipients, screening before and after transplantation and an individualized choice of immunosuppression are thus mandatory. PMID- 12584470 TI - Nomograms are superior to staging and risk grouping systems for identifying high risk patients: preoperative application in prostate cancer. AB - PURPOSE OF REVIEW: We outline a generic approach to using a nomogram to predict a continuous probability of failure in high-risk patients (rather than putting patients into groups), in order to identify patients whose risk exceeds a cutoff point. We discuss the goals of any staging system, what markers should be included, and models of markers. RECENT FINDINGS: Selection of high-risk patients for any cancer has traditionally been accomplished by the creation of risk groups, or perhaps clinical stages. Ideally, high-risk patients should be identified as accurately as possible, because of the treatment and psychological implications for the patient. We argue that a continuous multivariable prediction model, such as a nomogram, is the most appropriate and accurate way to select high-risk patients. This type of model predicts outcome more accurately than risk grouping or staging systems. As an example, we use our preoperative prostatic specific antigen recurrence nomogram to identify patients at high risk of biochemical failure, who are in need of an effective neoadjuvant therapy. SUMMARY: It will follow from our discussion that identification of high-risk patients should follow four simple steps. First, select the endpoint of interest for the trial or the patient. Second, select the method that predicts the endpoint as accurately as possible. Third, determine the cutoff of predicted probability beyond which it makes sense to give the patient experimental therapy. Fourth, offer the novel therapy to the patient whose prediction of the endpoint, using the most accurate prediction method, exceeds the threshold. PMID- 12584471 TI - Adjuvant external radiation therapy following radical prostatectomy for node negative prostate cancer. AB - PURPOSE OF REVIEW: The use of adjuvant radiation therapy following prostatectomy is commonplace. The purpose of this review is to summarize completed and ongoing clinical trials and to review recent relevant studies and debates related to this subject. RECENT FINDINGS: The routine use of adjuvant radiation therapy remains a controversial topic. Recent retrospective matched-pair analyses support its use in appropriately selected patients with positive margins, extraprostatic extension or seminal vesicle invasion, but interpretation of these and other data vary. Although the 5-year biochemical recurrence rate using adjuvant radiotherapy may be decreased from approximately 40 to 10% in patients with either positive margins or extraprostatic extension, its effect on cause-specific mortality is unclear. Two prospective randomized trials with cumulative enrollment of over 1400 patients have examined the role of adjuvant radiation therapy compared with observation following prostatectomy: one trial was a National Cancer Institute sponsored Intergroup study coordinated by the Southwest Oncology Group, and the other was from the European Organization for Research and Treatment of Cancer. Currently, the Radiation Therapy Oncology Group is conducting a three-arm trial, with broadened stratification criteria as compared with previous trials. This ongoing trial examines the use of adjuvant radiotherapy with or without adjuvant androgen deprivation following prostatectomy and also androgen deprivation alone in patients at high risk for disease relapse. SUMMARY: In lieu of data from completed randomized trials, indications for immediate adjuvant radiation therapy following prostatectomy exist and are supported by retrospective data with respect to reducing local and biochemical recurrence rates. However, data demonstrating an overall or cause-specific survival advantage for adjuvant radiotherapy as compared with delayed salvage therapy do not exist. PMID- 12584472 TI - The rationale for adjuvant chemotherapy for high-risk prostate cancer. AB - PURPOSE OF REVIEW: With the advent of prostate-specific antigen, stage migration has resulted in a shift towards early-stage prostate cancer at diagnosis. Although radical prostatectomy and radical radiotherapy can be curative in organ confined disease, there remains a significant proportion of early-stage patients who go on to develop progressive, incurable disease. This review will highlight developments in the identification of high-risk patients, and summarize the results of investigations of adjuvant chemotherapy in this setting. RECENT FINDINGS: The ability to identify patients at high risk of developing progressive disease is improving. Both preoperative and postoperative variables, as well as newer radiographic and molecular tools, can identify at-risk patients who may benefit from adjuvant therapy. Coupled with developments in chemotherapeutic agents for prostate cancer, this provides the rationale for investigating chemotherapy in this setting. Unfortunately, to date, reported trials involving adjuvant chemotherapy in prostate cancer are few, and generally involve small numbers of patients. Some of the studies confirm that certain populations of patients, such as those with node-positive disease, may benefit from systemic therapy. Definitive data, however, will be derived from ongoing randomized trials investigating adjuvant chemotherapy. SUMMARY: Although definitive data regarding systemic chemotherapy in adjuvant therapy are scarce, the results of the available studies, and the increasing accuracy in delineating the population at risk, have laid the foundation for future and ongoing studies in this area. PMID- 12584473 TI - The role of intravenous zoledronic acid in the management of high-risk prostate cancer. AB - PURPOSE OF REVIEW: There are limited effective therapies that would persuade most urologists actively to screen for and treat patients who have developed asymptomatic bone metastases. RECENT FINDINGS: Recently published trials regarding new treatment options for patients with high-risk metastatic prostate cancer are now available, which describe the utility of the intravenous bisphosphonate zoledronic acid. SUMMARY: Prostate cancer is the most common malignancy in men in the United States. Many prostate cancer patients, especially those with aggressive and advanced disease, are at increased risk of developing bone metastases and, subsequently, skeletal-related events. Skeletal-related events may result both from disease and/or treatment of disease, and include bone pain, pathological fractures and spinal cord compressions. Furthermore, bone radiation and surgery to address bone-related pain, prevent pathological fractures and/or spinal cord compression are also considered skeletal-related events. These sequelae of bone metastases are both painful and debilitating, and severely compromise a patient's quality of life. PMID- 12584474 TI - The use of complementary/preventive medicine to prevent prostate cancer recurrence/progression following definitive therapy: part I--lifestyle changes. AB - PURPOSE OF REVIEW: The number one cause of death in the United States and in most countries around the world is cardiovascular disease. The number one or number two cause of death in prostate cancer patients is also cardiovascular disease. These observations do not serve to belittle the impact of prostate cancer, but are a reminder that the ultimate goal of healthy lifestyle recommendations is to reduce the burden of both of these major causes of death, especially after definitive prostate therapy. Patients need to be encouraged to know their cholesterol levels and other cardiovascular markers including blood pressure, as well as being aware of their prostate-specific antigen values. RECENT FINDINGS: Patients should not smoke, they should reduce their intake of saturated and trans fats, increase their consumption of a diversity of fruit and vegetables, consume moderate quantities of dietary soy or flaxseed, increase their consumption of fish or fish oils and other omega-3 fatty acids, as well as maintaining a healthy weight, getting at least 30 min/day of physical activity, and lifting weights several times a week. When in doubt it is important for the clinician and patient to realize that what is healthy for the heart is generally found to be healthy for the prostate. Many of these lifestyle changes, when accomplished on a regular basis, may dramatically reduce the risk of overall early mortality. Despite the simplistic and moderate recommendations in this manuscript, research suggests that few individuals are currently following these suggestions. SUMMARY: Clinicians need to constantly emphasize these basic changes in order to truly impact the overall health of any patient following definitive prostate therapy. PMID- 12584475 TI - The use of complementary/preventive medicine to prevent prostate cancer recurrence/progression following definitive therapy. Part II--rapid review of dietary supplements. AB - PURPOSE OF REVIEW: The number one cause of death in the United States and in most countries around the world is cardiovascular disease. The number one or two cause of death in prostate cancer patients is also cardiovascular disease. These observations do not serve to belittle the impact of prostate cancer, but serve as a reminder that the ultimate goal of dietary supplement recommendations is to reduce the burden of both of these major causes of death, especially after definitive prostate therapy. Several supplements should be discussed with a patient following definitive prostate cancer therapy. RECENT FINDINGS: On the basis of observational studies, a general cheap multivitamin that provides the recommended daily values of folic acid, B6, B12 and vitamin D may reduce the risk of a variety of chronic diseases. Selenium supplements at 200 microg/day should be reserved only for men with a deficient level of plasma selenium, but healthy dietary sources should be recommended for most men. Low-dose vitamin E supplements may be adequate for current or recent smokers but not non-smokers. All men should be encouraged to consume healthy dietary sources of vitamin E, regardless of smoking status. Low-dose daily aspirin may be the best over the counter product to utilize as a preventive agent; this statement is made on the basis of the numerous clinical studies that support its use for individuals that qualify after consulting with their physician or a specialist to evaluate current cardiovascular risk. In addition, fish oil supplements have gained acceptance as a possible therapy in high-risk cardiovascular patients. SUMMARY: The potential future role in prostate cancer should be of interest, and preliminary data is noteworthy. Regardless, all of these supplements have indirect evidence for effects in prostate cancer, but it seems that only a minority of men overall qualify for them currently, with the exception of a low-dose aspirin or multivitamin. PMID- 12584476 TI - Quality of life following definitive therapy for localized prostate cancer: potential impact of multiple therapies. AB - PURPOSE OF REVIEW: Patients with clinically localized prostate cancer often undergo multiple therapies during their disease trajectory, either as planned combinations or as salvage for recurrence. Most studies of health-related quality of life in men with prostate cancer have focused on those receiving one modality or another. This review summarizes the little that is known about health-related quality of life after multiple therapies. RECENT FINDINGS: Twelve articles are reviewed. One is more of historical interest at this time and sets the stage for the great prevalence of combination therapy with androgen ablation in men undergoing pelvic irradiation. Six deal with brachytherapy combined with external beam radiation therapy, androgen ablation, or both. Three address androgen ablation or radiation for biochemical recurrence. One presents outcomes following salvage radiation, and one summarizes results from patients undergoing salvage radiation or prostatectomy following primary treatment with the other. SUMMARY: Combination therapy for early stage prostate cancer confers many additional decrements in health-related quality of life that must be considered when making treatment decisions, especially given the minimal differences in survival between various approaches. PMID- 12584479 TI - Complementary and alternative therapies in childhood attention and hyperactivity problems. AB - To determine the prevalence and factors associated with complementary and alternative medicine (CAM) use for childhood attention and hyperactivity problems, we surveyed parents of children referred for evaluation of attention deficit hyperactivity disorder (ADHD). Parents indicated whether they had used CAM therapies (e.g., acupuncture, nutritional supplements) in the past year and rated how important different reasons were in making their therapy decisions. Overall, 62 of 114 (54%) parents reported using CAM, most commonly expressive therapies, vitamins, and dietary manipulation, to treat their child's attention problems. Parents who used CAM rated a "natural therapy" and "having more control over treatments" significantly more important in their choice of therapy than parents who did not use CAM. Only 11% of parents discussed using CAM with their child's physician. Because parents often use CAM to treat their child's attention and hyperactivity problems without their pediatrician's knowledge, pediatricians need to initiate discussions of CAM use with patients and families. PMID- 12584480 TI - The impact on family scale revisited: further psychometric data. AB - This article presents additional psychometric data and a revised scoring procedure for the widely used Impact on Family Scale. The analyses were conducted with three datasets produced by validation studies conducted at the originating institution with populations drawn from the same inner-city communities as the original sample. The instrument has one robust factor representing overall family impact. In addition, there are two subsidiary sets of items (financial impact and sibling impact) of possible interest to future researchers. The correlation of the previously published and new Total Score is .97. Data indicate that the Impact on Family Scale is an easily administered, reliable, and valid measure of a family member's perception of the effect of a child's chronic condition that can be used across diagnostic groups. The Impact on Family Scale fills a gap in the measurement of the psychological and social consequences of chronic disorders in childhood that can be useful in clinical and health services research. PMID- 12584481 TI - Effects of prenatal methamphetamine exposure on fetal growth and drug withdrawal symptoms in infants born at term. AB - To determine fetal growth and the incidence of withdrawal symptoms in term infants exposed to methamphetamine in utero, we retrospectively identified neonates whose mothers used methamphetamine during pregnancy and matched them to unexposed newborns. Exclusion criteria included multiple and preterm gestations. Although there were no differences in infant growth parameters between the methamphetamine-exposed and methamphetamine-unexposed neonates, methamphetamine exposure throughout gestation was associated with decreased growth relative to infants exposed only for the first two trimesters. In addition, there were significantly more small for gestational age infants in the methamphetamine group compared with the unexposed group. Methamphetamine-exposed infants whose mothers smoked had significantly decreased growth relative to infants exposed to methamphetamine alone. Withdrawal symptoms (as determined by a previously reported scoring system) requiring pharmacologic intervention were observed in 4% of methamphetamine-exposed infants. These preliminary findings indicate that methamphetamine use is associated with growth restriction in infants born at term. PMID- 12584482 TI - Sibling relationships and behavior after pediatric traumatic brain injury. AB - To evaluate long-term outcomes for siblings of children with traumatic brain injury (TBI), measures of sibling relationships and sibling behavior were collected an average of 4 years postinjury. The study sample included participants in a larger longitudinal study who had school-aged siblings, including 34 with severe TBI, 30 with moderate TBI, and 39 with orthopedic injuries not involving brain insult (ORTHO group). Group comparisons revealed more negative sibling relationships in families of children with TBI than in families of children in the ORTHO group, but only for mixed-gender sibling pairings. Behavior problems in children with TBI predicted both sibling relationships and sibling behavior problems. The findings indicate a need to monitor the adjustment of siblings and sibling relationships after TBI and to include siblings in family interventions. PMID- 12584483 TI - Effects of women's stress-elicited physiological activity and chronic anxiety on fetal heart rate. AB - This study examined the effects of pregnant women's acute stress reactivity and chronic anxiety on fetal heart rate (HR). Thirty-two healthy third trimester pregnant women were instrumented to monitor continuous electrocardiography, blood pressure, respiration, and fetal HR. Subjects completed the trait anxiety subscale of the State Trait Anxiety Index, then rested quietly for a 5-minute baseline period, followed by a 5-minute Stroop color-word matching task and a 5 minute recovery period. Fetal HR changes during women's recovery from a stressful task were associated with the women's concurrently collected HR and blood pressure changes (r =.63, p <.05). Fetal HR changes during recovery, as well as during women's exposure to the Stroop task, were correlated with their mothers' trait anxiety scores (r =.39, p <.05 and r = -.52, p <.01, respectively). Finally, a combination of measures of women's cardiovascular activity during recovery and trait anxiety scores accounted for two thirds of the variance in fetal HR changes during the same recovery period (r =.69, p <.001). The results from this study link changes in fetal behavior with acute changes in women's cardiovascular activity after psychological stress and women's anxiety status. This indicates that variations in women's emotion-based physiological activity can affect the fetus and may be centrally important to fetal development. PMID- 12584485 TI - Enhancing behavioral and social skill functioning in children newly diagnosed with attention-deficit hyperactivity disorder in a pediatric setting. AB - The objective of this study was to evaluate the effectiveness of an 8-week behavioral and social skill (BSS) class for children newly diagnosed with attention-deficit hyperactivity disorder (ADHD) and their parents, initiating stimulant treatment in primary care. The subjects were 100 children, aged 5 to 12 years, recently diagnosed with ADHD and treated with stimulant medication, and their parents or guardians. Eligible families were randomly assigned to an intervention group (IG: n = 59) or control group (CG: n = 41). The BSS function of each child was assessed using DuPaul's ADHD Parent Rating Scale (18-item) and Child Attention Profile (12-item) during blinded baseline and follow-up interviews. Parent discipline practice was assessed using a five-item inventory based on Likert-scale ratings during identical periods. Computerized pharmacy records were used to track psychostimulant use over time. IG children exhibited significantly lower parent-rated ADHD symptoms, whereas IG parents reported significantly better and more consistent discipline practices compared with CG parents across time. No significant differences were found between groups in Child Attention Profile scores across time. Psychostimulant use did not significantly differ between groups across time. BSS training was well accepted and seemed to significantly improve BSS functioning among IG children in the home setting only. PMID- 12584484 TI - Influence of prenatal cocaine exposure on early language development: longitudinal findings from four months to three years of age. AB - The influence of prenatal cocaine exposure on children's language functioning was evaluated longitudinally at six time points from 4 months to 3 years of age. The Miami Prenatal Cocaine Study prospectively enrolled 476 full-term African American infants at birth, categorized as cocaine-exposed (n = 253) or non cocaine-exposed (n = 223) by maternal self-report and bioassays (maternal/infant urine, meconium). The Bayley Scales of Infant Development, scored using the Kent Scoring Adaptation for language, was administered at 4, 8, 12, 18, and 24 months. The Clinical Evaluation of Language Fundamentals-Preschool was administered at 3 years. In longitudinal analyses using Generalized Estimating Equations, cocaine exposed children had lower overall language skills than non-cocaine-exposed children (D = -0.151; 95% CI = -0.269, -0.033; p =.012). Longitudinal findings remained stable after evaluation of potential confounding influences including other prenatal substance exposures and sociodemographic factors. Preliminary evidence also indicated possible mediation through an intermediary effect involving cocaine-associated deficits in fetal growth. PMID- 12584486 TI - Families matter--even for kids in child care. PMID- 12584487 TI - International adoption: a four-year-old child with unusual behaviors adopted at six months of age. PMID- 12584488 TI - From neurons to neighborhoods: old and new challenges for developmental and behavioral pediatrics. AB - Rapidly advancing research in neurobiology and the behavioral and social sciences, coupled with dramatic changes in the social and economic circumstances under which families are raising young children, has created a highly dynamic context for early childhood policy, service delivery, and child rearing in the United States. A recent report from the National Research Council and the Institute of Medicine, presents a critical analysis of that complex knowledge base and a set of recommendations designed to close the gap between what we know and what we do to promote the healthy development of young children. This article examines the report through the lens of developmental and behavioral pediatrics and highlights both exciting opportunities and sobering challenges. PMID- 12584489 TI - Modern approach to children with osteogenesis imperfecta. AB - Osteogenesis Imperfecta (OI) is characterized by bone fragility. At least seven discrete types have been described ranging from mild disease to a lethal form. In a large number of cases, mutations in one of the two genes encoding type I collagen have been found. In forms recently described (types V, VI, VII), such mutations have been excluded. In two other forms, (Bruck, and osteoporosis - pseudoglioma syndromes) defects in other proteins have been characterized. In OI, bone fragility stems from: decreased bone mass, disturbed organization of bone tissue, and altered bone geometry (size and shape). Histologic studies have shown that increased bone turnover is the rule in OI bone. This justifies using bisphosphonates in order to reduce osteoclast mediated bone resorption. Initial results are encouraging. Cyclical intravenous pamidronate administration reduces bone pain and fracture incidence, and increases bone density and level of ambulation, with minimal side effects. Effects on bone include increase in size of vertebral bodies and thickening of cortical bone. These results allow for more efficacious corrective surgery using intramedullary rodding of the long bones and paravertegral instrumentation. Specific occupational and physiotherapy programs are integral parts of the treatment protocol. This multidisciplinary approach will prevail until strategies aiming at the correction of the basic defect(s) will have come to fruition. PMID- 12584490 TI - Osteogenesis imperfecta and hyperplastic callus formation in a family: a report of three cases and a review of the literature. AB - Osteogenesis imperfecta is one of the most common groups of inherited disorders of connective tissue. Hyperplastic callus formation in patients with osteogenesis imperfecta after fracture or surgery is a rare occurrence that has often been misdiagnosed as osteosarcoma. Previous series reported that hyperplastic callus formation is more often present in osteogenesis imperfecta male patients, with white sclerae, and a negative family history of the disorder. This is the first time that this complication has been presented in three female siblings, with a positive family history of osteogenesis imperfecta type IV. An association between osteogenesis imperfecta type IV and hyperplastic callus formation is unclear. This association might, however, be a separate, specific subtype of osteogenesis imperfecta, with an unknown inheritance pattern. PMID- 12584491 TI - Slipped upper femoral epiphysis. A retrospective, clinical and radiological study of fixation with a single screw. AB - We conducted a retrospective, clinical and radiological study of fixation with a single screw in slipped upper femoral epiphysis. Forty children (53 hips) who had a slipped upper femoral epiphysis were managed by in-situ pinning with a single cannulated screw. The average age of these patients was 12.7 years and their average weight was above the 50th centile at the time of diagnosis. There were three acute slips, eight acute-on-chronic slips and 35 chronic slips. They were followed for an average of 2 years. Fifty-eight percent of patients had an excellent clinical result, 36% a good result, 4% a fair result and 2% a poor result. The complication rate was 5.3%. Our experience with single cannulated screw fixation of slipped upper femoral epiphysis is encouraging. No complications resulted from the choice of the fixation device. PMID- 12584492 TI - Connecting bar for hip spica reinforcement: does it help? AB - The purpose of this prospective study was to determine if a connecting bar prevented mechanical failure of hip spicas. Eighty-two hip spicas were applied to 70 patients between April 2000 and June 2001 following open or closed reduction for developmental dysplasia of the hip. Bilateral hip surgery was undertaken in 12 patients. Thirty-six spicas were applied with a bar whilst 46 were without. The technique of application was similar in all patients. Overall nine (11%) hip spicas failed (mechanically) prematurely and had to be replaced in hospital under general anaesthesia. All nine were inpatients whose spica did not have a connecting bar (P < 0.005). The rate of spica revision increased with the age of the patient. All parents of failed spica cases (nine cases) preferred the new spica with a bar to the previous one without, because they agreed that the bar made the spica more secure. The bar did not hamper toileting or handling. PMID- 12584493 TI - Development of hip dysplasia in a child with perinatal contralateral high above knee amputation. AB - The development of acetabular dysplasia in the growing child has been postulated to be due to abnormal forces on the acetabular rim, such as when the hip is in an adducted position. In this case report we document the development of acetabular dysplasia, as measured radiographically, in a child with a perinatal contralateral high above-knee amputation. PMID- 12584494 TI - Present status of screening and prevention of developmental dysplasia of the hip in the Slovak Republic. AB - The authors analysed the status of screening and prevention of developmental dysplasia of the hip in the Slovak Republic on the basis of the European Pediatric Orthopaedic Society questionnaire. The questionnaire was sent to 31 heads of orthopaedic departments, with 23 (74%) responses. According to the responses received, in the Slovak Republic a neonatologist or orthopaedic surgeon, depending on availability, makes the first clinical examination of the hips up to the fifth day after delivery. Every instability, relative shortening of one of the lower limbs, or 'high-risk' baby is referred immediately to the orthopaedic surgeon. Sonographic examination up to 7 days after delivery is performed in six departments. In another 11 departments, two or more sonographic examinations are performed on each baby. Authors support the simple static ultrasound as an effective screening test for developmental dysplasia of the hip, that should be applied to the whole population and not simply to the at risk group. PMID- 12584495 TI - Reduction versus remodeling in pediatric distal forearm fractures: a preliminary cost analysis. AB - The inherent ability of pediatric metaphyseal radius fractures to heal and remodel made us question the need for immediate anatomic reduction under conscious sedation. We believe that isolated closed distal radius fractures with 15 degrees of angulation and 1 cm of shortening will heal well and remodel completely without clinical or functional sequelae. Time and expense can be decreased by splinting and follow-up without the need for immediate anatomic reduction in the emergency room. In order to answer this question, we retrospectively evaluated 34 pediatric metaphyseal wrist fractures that lost position after attempted reduction and healed in their angulated or shortened position. We looked at the time to healing, time to remodeling and any residual clinical or functional deficits. We then did a comparison cost analysis with time matched patients who had complete but minimally displaced fractures of the distal radius that were treated by immediate splinting with orthopaedic follow-up. Our results showed that skeletally immature patients with open physes, isolated injuries, dorsovolar and radioulnar angulations less than 15 degrees and less than 1 cm of shortening will heal and be out of cast within an average of 6 weeks and completely remodel within an average of 7.5 months. The average time in the emergency room was 2 h less with no reduction. The cost of the emergency room visit with attempted reduction was 50% more than splinting with early referral (US dollars 536 versus US dollars 270). None of our patients had significant clinical deformities or residual functional deficits. PMID- 12584496 TI - Supracondylar medial open wedge osteotomy with external fixation for cubitus varus deformity. AB - BACKGROUND: Many surgical procedures are used for the correction of cubitus varus deformity in children. Even if the primary indication is cosmetic, but many poor results have been described. The technique of a supracondylar medial opening wedge osteotomy with external fixation is presented and preliminary results in four cases are reported. TECHNIQUE: An antero-medial approach is chosen with exposition and anterior transposition of the ulnar nerve. Under image intensifier control, insertion of four Schanz screws is performed from medial to lateral. In between, an incomplete medial osteotomy is performed obliquely (in the antero posterior as well as in the medio-lateral plane), leaving a small part of the radial humeral cortex intact. The osteotomy is opened until varus and, if present, hyperextension deformation are corrected. Then, the fixator system is applied. RESULTS: All osteotomies healed primarily. Mean valgus correction of 21.75 degrees was achieved; in only one patient additional flexion modification of 17 degrees was necessary. At a follow-up of at least 2 years clinically three out of four presented symmetric elbow position, one slight residual varus deformity was noted in spite of 20 degrees of valgization due to important valgus position of the contralateral side. No lateral bulging of the corrected elbow was observed. CONCLUSION: The described technique is an alternative to other procedures, with special regard to the cosmetic outcome as well as to the control of correction for valgus and flexion. PMID- 12584497 TI - Lower limb deformities in children: two-stage correction using the Taylor spatial frame. AB - The Taylor spatial frame is a new external ring fixation system developed for correction of axial, sagittal, planar and rotational deformities of the extremities. Between August 1999 and October 2000 five children were treated by this system. Four had shortened and deformed femurs after fracture, osteomyelitis or a congenital short femur. One patient had a pseudoarthrosis of the tibia. Mean corrections were leg lengthening 5.9 cm (1.7-7.2), valgus correction 10.6 degrees (0-19), lateral translation 12.6 mm (0-33) and rotation 10 degrees. The mean healing index was 48.4 days/cm. In all patients the bones fused without major complications. We recommend the Taylor spatial frame for corrections of complex limb deformities. PMID- 12584498 TI - Physeal arrest of the ankle secondary to extravasation in a neonate and its treatment by the Gruca operation: a modern application of an old technique. AB - We report a case of physeal arrest secondary to extravasation of an intravenous infusion in a neonate. The ankle was reconstructed according to the Gruca operation followed by callus distraction lengthening. PMID- 12584499 TI - A method for the early evaluation of the Ponseti (Iowa) technique for the treatment of idiopathic clubfoot. AB - The Ponseti casting technique is reported to have a high success rate in the treatment of idiopathic clubfoot. Non-operative treatment of clubfoot provides a lower complication rate, less pain, and higher function as the patient ages than operative treatment. To demonstrate serial post-treatment change in clubfeet over time, three clubfoot rating systems were utilized in the current study. Patients compliant with the Ponseti technique and treated before the age of 7 months, had a 92% success rate at an early follow-up after casting was completed. It is not the purpose of this article to analyze the long-term clubfoot treatment result but to establish tools which can be used to judge initial success with the Ponseti technique. Complications are few and minor, limited to equipment used and cast technique. PMID- 12584500 TI - The development in footprint morphology in 1851 Congolese children from urban and rural areas, and the relationship between this and wearing shoes. AB - The footprints of 1851 Congolese children were studied using the index of Chippaux-Smirak, Staheli's index of the arch, and Clarke's angle. The sample consisted of 906 girls and 945 boys aged between 3 and 12 years, including city children who wore shoes and children from rural areas who had gone barefoot. At the ages of 3 and 4 years, most feet were morphologically flat, but the proportion of flat feet decreased with age in both sexes. Boys had a greater tendency for flat feet. According to the three parameters studied there was a greater proportion of flat feet in the urban environment. By multiple regression analysis the three parameters we had studied identified age as the primary predictive factor for flat feet. As in previous studies in western populations, the girls had a higher inner arch than the boys and footwear had very little influence on the morphology of the foot. PMID- 12584501 TI - Maffucci lymphangioma syndrome: an unusual variant of Ollier's disease, a case report and a review of the literature. AB - A case of Maffucci's syndrome in an 18-month old boy is presented, in which the vascular malformation consists of lymphangioma. This is a rare variant of Maffucci's syndrome. So far only six other cases have been reported in which lymphangiomas featured. We believe that the case described here is only the second case to be reported for which a lymphangioma is the only vascular malformation in the syndrome. The literature of the other six cases was reviewed and various features identified enabling this particular disease to be classified as a distinct variant of Maffucci's syndrome. PMID- 12584502 TI - Atypical mycobacterial granulomatous epiphyseal osteomyelitis in an immunocompetent child. AB - Osteomyelitis caused by atypical mycobacteria is rare in children. The majority of affected patients have had some kind of predisposing factor, namely a penetrating injury or surgery, or were immune-compromised. Our experience shows that this diagnosis should be considered in apparently healthy children as well. The use of polymerase chain reaction has now made it possible to identify the pathogen in cases that were previously diagnosed as granulomatous osteomyelitis. We present a case of atypical mycobacterial osteomyelitis affecting the distal femoral epiphysis in an immunocompetent 10-year-old child. The diagnosis in this case was made by the use of the polymerase chain reaction assay. PMID- 12584503 TI - The stubborn hip: idiopathic avascular necrosis of the hip. AB - OBJECTIVE: To highlight the unusually indolent course of avascular necrosis in this patient, the risk factors of which chiropractors should be aware, the necessity of and means to an early diagnosis, the limitations of plain film radiographs, as well as the current medical treatments. CLINICAL FEATURES: A 27 year-old male professional soccer player had a tight and achy right hip for approximately 6 years, a condition that increased with physical activity. His active range of motion was limited by 25% in extension and abduction, and all resisted ranges of motion produced pain. After an equivocal radiograph and bone scan, magnetic resonance imaging revealed a subchondral defect located on the superior aspect of the right femoral head, consistent with avascular necrosis of the femoral head. INTERVENTION AND OUTCOME: Presurgical management included Cybex testing, massage, myofascial release, interferential current, muscle strengthening, and muscle balancing exercises 3-5 times per week for 5 months. The patient underwent a curette procedure. Six weeks later, he returned to the chiropractic office for postsurgical rehabilitation, in which he is still involved today. He has made progress with respect to flexibility, strength, and muscle coordination. CONCLUSIONS: Idiopathic avascular necrosis of the hip is a clinical entity characterized by pain in the hip made worse with activity and at night, which may radiate to the groin, back, or thigh. Often, physical exam and radiographic findings are equivocal, at which point advanced imaging such as magnetic resonance imaging should be ordered to identify the disease in its earliest stages. PMID- 12584504 TI - Danish chiropractic patients then and now--a comparison between 1962 and 1999. AB - BACKGROUND: Little is known about changes in the chiropractic patient population over time. OBJECTIVE: To compare 2 surveys describing the Danish chiropractic patient population. DESIGN: Data concerning location of primary complaint and its duration for patients in Danish chiropractic offices between 1962 and 1999 were compared. SETTING: Private chiropractic practice and nonprofit research institution. OUTCOME MEASURES: Location of primary diagnosis/complaint, duration of complaint. RESULTS: In 1962, data for 1118 patients were collected, with a participation rate of 93%. In 1999, data for 1897 patients were collected. Of all Danish chiropractic clinics, 88% participated in the study, and 94% of all eligible patients filled out a questionnaire. In both 1962 and 1999, the most frequent complaints were pain in the lower back or neck either alone or with radiation to the extremities (roughly 70% of patients). In 1962, almost 4 times as many patients complained of headache as in 1999 (11% vs 4%). Although there were significant differences, less that 10% of patients presented with a nonmusculoskeletal disorder in both 1962 and 1999 (7% vs 3%). In 1962, almost half of the patients had had their complaint for more than 1 year; in 1999, approximately 80% of patients had had their complaint for less than 1 year. CONCLUSIONS: In both 1962 and 1999, Danish chiropractors primarily treated patients with pain syndromes related to the lower back and neck. Patients presenting with type O disorders comprised less than 10% of the total patient population in both surveys. PMID- 12584505 TI - Do chiropractic colleges' off-campus clinical sites offer diverse opportunities for learning? A preliminary study. AB - OBJECTIVE: This study involved the collection of data on the demographic characteristics and chief complaints of patients of chiropractic college teaching clinics to study whether patients who visited different types of teaching clinics were demographically or clinically dissimilar. It represents a first step toward our ability to collect data from teaching clinics to test for differences between specific clinic populations. DESIGN: Descriptive cross-sectional study, implemented through a survey of patients. A survey form and a data collection infrastructure already in place for an established practice-based research program allowed the application of existing resources in the teaching clinic environment. PARTICIPANTS: Participating clinics included on-campus, off-campus, and outreach teaching clinics of four chiropractic colleges. Recruitment took place by word of mouth; clinic directors volunteered to participate in the 1-week survey. The study population consisted of all patients, new and established and of any age, who came to participating clinics during the week of March 16-21, 1998. RESULTS: Fourteen participating clinics returned surveys on 1612 patients. The age, sex, and race of patients, and the nature, onset, and duration of their chief complaints were significantly different among the categories of teaching clinics. Salient dissimilarities occurred in racial distribution, types of chief complaint, and duration of chief complaint among the patient populations of different clinics. CONCLUSION: Different types of teaching clinics may facilitate greater diversity in the patient population seen by chiropractic students, broadening the clinical training of chiropractic students by providing diversity in clinical settings and patient populations. PMID- 12584506 TI - Responsiveness of the Bournemouth and Oswestry questionnaires: a prospective pilot study. AB - BACKGROUND: The assessment of health-related quality-of-life outcome information has become an essential feature in the delivery of quality health care. Outcome assessment and increased clinical research capacity have been identified as recommended areas of priority for chiropractic research, chiropractic colleges, and the profession. OBJECTIVE: This article reports the results of a multisite pilot project utilizing New York Chiropractic College's 3 off-campus health centers. In the project, we sought to examine generic issues related to research infrastructure and feasibility, perform pilot testing of the responsiveness of the revised Oswestry Disability Index (ODI) and the Bournemouth Questionnaire (BQ), and develop estimates of change and population size for the possible future projects. METHODS: We describe the infrastructure created to conduct the project, including an on-site clinician research manager; on-site student research assistants; identification, recruitment, and patient flow; demographic and clinical characteristics; and abstraction instruments. Responsiveness study subjects completed 3 health-related quality-of-life assessments at baseline, 15 days, and 30 days. At 45 days, or at discharge, subjects completed the same questionnaires and a global rating of change. Responsiveness is reported by analysis of standardized estimates of change, relative efficiency, and receiver operator characteristic (ROC) curve plots. RESULTS: We successfully identified 82% of possible study patients; the real eligibility rate in our population was 30%. In 4 months, 70 patients were accrued, and 26 patients followed through to provide global ratings of change scores. Responsiveness by standardized change scores was numerical rating scale (NRS) = 1.19, ODI = 0.91, BQ = 0.78; by relative efficiency it was BQ/ODI = 0.86, ODI/NRS = 0.76, BQ/NRS = 0.66. Area under the curves ranged from 0.69 for the BQ to 0.86 for the NRS. CONCLUSIONS: Recommendations for modification and use of the infrastructure, research capacity, and future outcome and appropriateness assessment projects are offered. PMID- 12584507 TI - Repeatability over time of posture, radiograph positioning, and radiograph line drawing: an analysis of six control groups. AB - BACKGROUND: There is debate concerning the repeatability of posture over time, radiograph positioning repeatability, and radiograph line drawing reliability. These ideas seem to negate the use of before-and-after spinal radiographic imaging to detect and correct vertebral subluxations. OBJECTIVE: To review the results of control groups in 6 clinical control trials with before-and-after radiographic measurements taken days, weeks, months, or years apart to accept or reject the hypothesis that radiographic analysis procedures are not repeatable, reliable, or reproducible. DATA SOURCES: Six published control groups from original data. Other data were obtained from searches on MEDLINE, CHIROLARS, MANTIS, and CINAHL on radiographic reliability, posture, and positioning. RESULTS: Comparison of initial and follow-up radiographic data for 6 control groups indicate that measured angles and distances between initial and follow-up radiograph measurements on lateral and anterior to posterior radiographs are not significantly different when utilizing Chiropractic Biophysics radiographic procedures. In 48 out of 50 measurements, the differences between initial and follow-up radiographs are less than 1.5 degrees and 2 mm. These measurements indicate that posture is repeatable, radiographic positioning is repeatable, and radiographic line drawing analysis for spinal displacement is highly reliable. The scientific literature on these topics also indicates the repeatability of posture, radiographic positioning, and radiographic line drawing. CONCLUSIONS: Posture, radiographic positioning, and radiographic line drawing are all very reliable/repeatable. When Chiropractic Biophysics standardized procedures are used, any pre-to-post alignment changes in treatment groups are a result of the treatment procedures applied. These results contradict common claims made by several researchers and clinicians in the indexed literature. Chiropractic radiologic education and publications should reflect the recent literature, provide more support for posture analysis, radiographic positioning, radiographic line drawing analyses, and applications of posture and radiographic procedures for measuring spinal displacement on plain radiographs. PMID- 12584508 TI - Aberrant protective force generation during neural provocation testing and the effect of treatment in patients with neurogenic cervicobrachial pain. AB - BACKGROUND: Observation of the occurrence of protective muscle activity is advocated in assessment of the peripheral nervous system by means of neural provocation tests. However, no studies have yet demonstrated abnormal force generation in a patient population. OBJECTIVES: To analyze whether aberrations in shoulder girdle-elevation force during neural tissue provocation testing for the median nerve (NTPT1) can be demonstrated, and whether possible aberrations can be normalized following cervical mobilization. STUDY DESIGN: A single-blind randomized comparative controlled study. SETTING: Laboratory setting annex in a manual therapy teaching practice. PARTICIPANTS: Twenty patients with unilateral or bilateral neurogenic cervicobrachial pain. METHODS: During the NTPT1, we used a load cell and electrogoniometer to record continuously the shoulder-girdle elevation force in relation to the available range of elbow extension. Following randomization, we analyzed the immediate treatment effects of a cervical contralateral lateral glide mobilization technique (experimental group) and therapeutic ultrasound (control group). RESULTS: On the involved side, the shoulder-girdle elevation force occurred earlier, and the amount of force at the end of the test was substantially, though not significantly, greater than that on the uninvolved side at the corresponding range of motion. Together with a significant reduction in pain perception after cervical mobilization, a clear tendency toward normalization of the force curve could be observed, namely, a significant decrease in force generation and a delayed onset. The control group demonstrated no differences. CONCLUSIONS: Aberrations in force generation during neural provocation testing are present in patients with neurogenic pain and can be normalized with appropriate treatment modalities. PMID- 12584509 TI - Chiropractic management of chronic chest pain using mechanical force, manually assisted short-lever adjusting procedures. AB - OBJECTIVE: To discuss a case involving a patient with chronic chest pain, dyspnea, and anxiety. Although resistant to previous treatment regimens, the condition responded favorably to chiropractic manipulation of the costosternal articulations. CLINICAL FEATURES: A 49-year-old man had chronic chest pain, dyspnea, and anxiety for over 4 months. The severity of the condition gradually progressed to the point of precluding the patient's active employment and most physical activity. Prior efforts to treat the condition had met with failure. INTERVENTION AND OUTCOME: The patient received mechanical force, manually assisted short-lever chiropractic adjustment of the thoracic spine and, in particular, the costosternal articulations. Adjustments were by means of an Activator Adjusting Instrument II. The patient responded favorably to the intervention, obtaining prompt relief from his symptoms. Sustained chiropractic care rendered over a 14-week period resulted in complete resolution of the patient's previously chronic condition, with recovery maintained at 9-month follow-up. CONCLUSIONS: Certain types of chest pain may have their etiology in a subluxation complex involving the costosternal articulation. Although the possibility of myocardial involvement must be considered with all patients whose symptoms include chest pain, a musculoskeletal involvement, including costosternal subluxation complex, may be the underlying cause of the symptoms in certain patients. When this is the case, chiropractic adjustment may provide an effective mode of treatment. Further study in an academic research venue is merited to investigate the role that conservative chiropractic care can provide for patients with chest pain. PMID- 12584510 TI - Neurofibromatosis type I: clinical and imaging features of Von Recklinghausen's disease. AB - OBJECTIVE: To discuss the case of a 45-year-old woman, with a prior diagnosis of neurofibromatosis type 1 (NF-1), complaining of low back and cervical spine pain with bilateral upper extremity paresthesias. CLINICAL FEATURES: The patient had a dull, achy, constant low-back pain of 4 months' duration, with mild headaches and upper extremity paresthesias. Multiple skin lesions and spinopelvic postural imbalances were present. Diagnostic radiography, along with advanced imaging, demonstrated multilevel dysplastic osseous changes, with dural ectasia, scoliosis, and tumor extension. In addition to the previous diagnosis of NF-1, our clinical diagnosis included segmental dysfunction with resultant cervicalgia, lumbalgia, and myospasm. INTERVENTION AND OUTCOME: Treatment consisted of a course of spinal manipulation of the lumbopelvic region, with adjunctive therapy consisting of interferential therapy, heat, and rehabilitative exercise. Marked reduction in pain and paresthesia with improved function were achieved. CONCLUSIONS: NF-1 is a multisystem disease with neoplasia of the skin and nervous system. Patients experience a lifetime of morbidity and increased risk of mortality, depending on the extent of the disease. A multitude of therapeutic regimens may be engaged to improve NF-1-associated symptomatology and morbidity. Chiropractic spinal manipulation may have a positive effect in pain reduction and improved function in patients who have NF-1 without spinal instability. PMID- 12584511 TI - A comparison of methods of evaluating cervical range of motion. PMID- 12584513 TI - Viruses and asthma: Inception, exacerbation, and possible prevention. AB - Viral respiratory tract infections have been epidemiologically associated with asthma in at least 3 ways. First, during infancy, certain viruses have been implicated in the inception of the asthmatic phenotype. Second, in patients with established asthma, particularly children, viral upper respiratory tract infections play a significant role in producing acute exacerbations of airway obstruction, often resulting in outpatient visits or hospitalizations. This increased propensity of viral infections to produce lower airway symptoms in asthmatic persons may be related, at least in part, to interactions among allergic sensitization, allergen exposure, and viral infections-all acting as cofactors in the induction of acute episodes of airflow obstruction. Third, and perhaps counterintuitively, certain infections may actually prevent the development of allergic respiratory tract diseases, including asthma. This review discusses these associations as they pertain to both the pathogenesis and treatment of childhood asthma. PMID- 12584514 TI - Mechanisms of virus-induced asthma. AB - In children and adults with asthma, infections with common cold viruses such as respiratory syncytial virus and rhinovirus frequently cause exacerbations of asthma. Recent studies suggest that these and other respiratory viruses infect the lower airways, and that the infected epithelial cells help to initiate immune responses to the virus. These immune responses have antiviral effects, but also enhance airway inflammation and may add to cold symptoms and airway obstruction. Moreover, there are data to suggest that there may be specific interactions between allergic and virus-induced inflammatory responses that contribute to lower airway dysfunction in asthma. Finally, impaired T-helper type 1-like lymphocyte responses during viral infections may be a risk factor for more severe viral illnesses in patients with allergies and asthma. It is hoped that these advances in our understanding of the relationship between viral infections and exacerbations of asthma will translate into new therapeutic targets for virus induced exacerbations of asthma. PMID- 12584515 TI - Clinical patterns and natural history of asthma. AB - Childhood asthma typically begins in infancy with a respiratory syncytial virus (RSV) infection. Although the majority of infants become infected with RSV, lower respiratory illness develops in only about 20%. About 25% to 50% of those subsequently experience recurrent acute asthma from viral respiratory infections (VRI). Children younger than 5 years have a high frequency of VRI and have the highest frequency of hospitalization for asthma of any age group. In a 35-year study of the natural history of asthma, 20% of 7-year-old children were found to have asthma, but most had only episodic illness with VRI. The majority of those children improved with age, but a substantial minority continued to have recurrent episodes as adults, generally induced by VRI or exercise. Persistent asthma developed in only a few. Children who had symptoms of asthma without VRI were more likely to continue having frequent episodic or chronic asthma as adults. Despite generally suboptimal treatment during the 35 years of the study, forced expiratory volume at one second did not deteriorate over time; it remained normal in children who had only episodic asthma, and it was consistently low in the children with severe, persistent asthma. PMID- 12584516 TI - Limitations of maintenance therapy for viral respiratory infection-induced asthma. AB - In asthmatic children with persistent symptoms, maintenance therapy with nedocromil sodium, inhaled corticosteroids, or montelukast is associated with a decreased rate of asthma exacerbations. The greatest benefit is seen with use of regular inhaled corticosteroids in preschool- or school-age children, in whom asthma exacerbations, the need for rescue oral corticosteroids, unscheduled urgent medical visits, and hospitalizations are all decreased by approximately 50%. However, maintenance therapy is not beneficial in children with intermittent respiratory virus-induced wheezing without persistent symptoms. PMID- 12584517 TI - Are inhaled corticosteroids effective for acute exacerbations of asthma in children? AB - If started early and at high doses, the acute use of inhaled corticosteroids (ICS) provides modest benefit in patients with mild exacerbations of asthma. In more severe exacerbations, oral corticosteroids are significantly more effective in preventing hospitalizations and improving lung function. We conclude that ICS should not be used routinely to treat acute exacerbations. PMID- 12584518 TI - Treatment strategies for viral respiratory infection-induced asthma. AB - The many therapeutic options for asthma can confuse both physicians and patients. Great emphasis has been placed on maintenance medication, particularly inhaled corticosteroids (ICS), which are the most effective medication for patients with persistent symptoms. However, asthma in young children is most commonly intermittent, triggered almost exclusively by viral respiratory infections (VRI). It is, nevertheless, associated with the highest rate of hospitalization of any age. Conventional doses of ICS do not prevent exacerbations of VRI-induced asthma. In contrast, intervention with oral corticosteroids during exacerbations has been shown to prevent the progression that would otherwise require urgent care or hospitalization. An oral corticosteroid kept on hand by the patient permits initiation of more prompt and effective treatment than is likely to occur when a patient must first go to a physician's office or emergency department, because it can be given as soon as the response to bronchodilator therapy is incomplete. Monitoring by phone and regularly scheduled physician visits to review the clinical course and reinforce education can then replace urgent medical care, thereby improving the quality and efficacy of asthma management. PMID- 12584519 TI - Selecting a systemic corticosteroid for acute asthma in young children. AB - Oral corticosteroids are as effective as intravenous therapy for treating acute exacerbations of asthma. They are available in tablets that can be crushed and mixed with soft food or syrup, and in a variety of liquid formulations that differ in volume required, palatability, patient acceptance, and cost. The most important consideration in product selection for a young child is that the doses can be easily swallowed and retained. PMID- 12584520 TI - Consensus statement from a conference on treatment of viral respiratory infection induced asthma in young children. PMID- 12584521 TI - Informed consent for pediatric research: is it really possible? PMID- 12584522 TI - Growth hormone for small-for-gestational-age children: short and sweet? PMID- 12584523 TI - While you were sleeping... PMID- 12584524 TI - Packed red blood cell transfusions for critically ill pediatric patients: when and for what conditions? PMID- 12584525 TI - Abnormal glucose tolerance in cystic fibrosis: why should patients be screened? PMID- 12584526 TI - The revolution in molecular biology leads to new understanding of the clinical expression of immunodeficiencies. PMID- 12584527 TI - When do the federal regulations allow placebo-controlled trials in children? PMID- 12584528 TI - Do healthy children deserve greater protection in medical research? PMID- 12584529 TI - Reduced insulin sensitivity during growth hormone therapy for short children born small for gestational age. AB - OBJECTIVES: To examine the influence of recombinant human growth hormone (rhGH) therapy on insulin sensitivity in short children born small for gestational age (SGA). STUDY DESIGN: Twelve short (height standard deviation score, -3.2 +/- 0.1) non-GH-deficient children SGA (7 boys/5 girls) were studied at 9.3 +/- 1.0 years of age. The insulin sensitivity index was measured with Bergman's minimal model before (11 children) and during (12 children) rhGH therapy (21 +/- 6 months) administered daily at 20 IU/m(2) per week. No child had a change in pubertal status during the study. In addition, 5 children who remained prepubertal had insulin sensitivity remeasured 3 months after rhGH therapy was suspended. RESULTS: With rhGH therapy, insulin sensitivity fell 44% +/- 10% (P =.018), with a compensatory rise in the acute insulin response of 123% +/- 59% (P <.009). Reassessment of insulin sensitivity in 5 children (3 boys/2 girls) 3 months after suspension of rhGH occurred at 9.9 +/- 0.7 years. Insulin sensitivity remained unchanged after rhGH therapy was stopped: 31.6 (20.5-42.3) before treatment, 11.5 (5.7-24.4) with treatment, and 10.7 (6.2-16.9) 10(-4). min(-1) microU/mL after treatment. CONCLUSIONS: Children SGA are known to have reduced insulin sensitivity. There was a further reduction in insulin sensitivity with rhGH therapy that did not recover 3 months after rhGH therapy was stopped. PMID- 12584530 TI - Day- and night-time blood pressure elevation in children with higher grades of renal scarring. AB - OBJECTIVE: We used 24-hour ambulatory blood pressure measurement (ABPM) to screen for early changes in children with renal scarring compared with healthy controls, and we investigated correlations between the extent of renal damage and blood pressure (BP). Study design Untreated patients (n = 61) with renal scars associated with recurrent urinary tract infection and vesico-ureteric reflux were investigated and compared with 904 healthy controls. RESULTS: A significant positive correlation between the extent of renal scarring and systolic and diastolic BP standard deviation score (SDS) was found by using ABPM. Mean systolic day-time BP SDS in ABPM was significantly elevated in girls with renal scars compared with healthy controls, but failed to be significant in boys. Mean systolic and diastolic night-time BP SDS were significantly elevated in patients. Compatible with these results, diastolic dipping was significantly lower in patients compared with controls. CONCLUSIONS: The increase in BP obtained by ABPM is positively correlated with the degree of scarring in our group of patients. Elevated night-time BP might be the most sensitive indicator of BP elevation in these children. PMID- 12584531 TI - Pediatric red blood cell transfusions increase resource use. AB - OBJECTIVE: To test the hypothesis that red blood cell transfusions are associated with increased resource utilization and mortality in critically ill children. METHODS: Five pediatric intensive care units (PICUs) participated in a retrospective, cohort analysis (1996-1999). Children with a hemoglobin value or=5 gpELISA units) after 2 doses. At 2-year follow-up, all patients retained detectable antibody, and 91% (21 of 23) had levels >or=5 gpELISA units. There were no adverse events associated with vaccination. CONCLUSIONS: Varicella vaccine was generally well tolerated and highly immunogenic in children with nephrotic syndrome, including those on low dose, alternate-day prednisone. PMID- 12584536 TI - Altered myocardial flow reserve and endothelial function late after Kawasaki disease. AB - OBJECTIVES: Coronary arterial lesions after Kawasaki disease (KD) may cause coronary endothelial dysfunction as the result of intimal hypertrophy. Our purpose was to assess myocardial flow reserve (MFR) and endothelial function in various myocardial regions after KD by using positron emission tomography. STUDY DESIGN: Twenty-seven patients, 17.2 +/- 3.2 years of age, who had KD at 1.9 +/- 1.4 years, and 12 normal healthy subjects, 26.5 +/- 3.4 years of age, were evaluated by means of myocardial blood flow (MBF) with (15)O-water positron emission tomography. MFR was estimated by MBF changes under adenosine triphosphate infusion and endothelial function by MBF changes under cold pressor testing. The left ventricle was divided into three coronary territories. Ten stenotic regions, 20 aneurysmal regions, 30 regressed aneurysmal regions, and 21 regions without coronary arterial lesions were compared with 36 control regions of the normal volunteers. RESULTS: MBF at rest was similar in each region. Hyperemic blood flow and MFR in each region after KD was significantly lower than those in the regions of normal volunteers. MBF during cold pressor testing was significantly reduced in each region after KD, as compared with no change in the control regions. CONCLUSIONS: Our study indicates impaired MFR and endothelial function regardless of coronary artery status after KD. PMID- 12584537 TI - Safety and efficacy of nonsteroid pimecrolimus cream 1% in the treatment of atopic dermatitis in infants. AB - OBJECTIVE: The safety and efficacy of a 1% cream formulation of pimecrolimus, a selective, nonsteroid immunomodulator, was studied in infants with atopic dermatitis (AD). METHODS: During a 6-week double-blind phase, 186 infants with mild/moderate AD were randomly assigned to twice-daily pimecrolimus cream 1% or vehicle. All patients were subsequently treated with open-label pimecrolimus for 20 weeks. RESULTS: At the end of the double-blind phase, 54.5% and 23.8% of patients in the pimecrolimus and vehicle groups, respectively, were clear or almost clear of AD (P <.001). Similar improvements were observed in the Eczema Area and Severity Index, pruritus assessment, and the care giver's assessment. By the first return visit, 69.9% and 36.5% of pimecrolimus and vehicle-treated patients, respectively, achieved absent or mild pruritus. Efficacy during the double-blind phase was maintained throughout the open-label phase. Vehicle treated patients transferring to open-label pimecrolimus rapidly achieved disease control comparable to those receiving continuous pimecrolimus. There were no significant differences between groups in application site reactions or skin infections. Most adverse events were mild or moderate and unrelated to treatment. CONCLUSIONS: Pimecrolimus was safe in infants with AD, with rapid and sustained efficacy. Pimecrolimus holds promise as a valuable new treatment option for the youngest patients with AD. PMID- 12584538 TI - Interactions between hypoglycemia and sleep architecture in children with type 1 diabetes mellitus. AB - OBJECTIVES: To assess the interactions between nocturnal hypoglycemia and sleep in children with type 1 diabetes mellitus (DM). STUDY DESIGN: Children with DM (n = 15) and 15 matched control children underwent full night polysomnographic recordings. Blood glucose levels were measured in the diabetic children by means of the MiniMed Continuous Glucose Monitoring System. Six of the diabetic children were also studied by peripheral arterial tonometry (an indirect indicator of sympathetic responses). RESULTS: Five children with DM (33%) had profound nocturnal hypoglycemia, which was associated with increased sleep efficiency, increased slow wave sleep, and increased Delta power in spectral analysis of the electroencephalogram. Hypoglycemic episodes were not associated with sympathetic activation. Rapid decline in glucose levels (>25 mg/dL/hour) but not the absolute degree of hypoglycemia were associated with awakenings from sleep. CONCLUSIONS: We conclude that sleep may inhibit sympathetic and arousal response to hypoglycemia. Rapid changes in glucose levels, independent of absolute glucose levels, may result in awakening from sleep. Continuous measurement of glucose levels during sleep may add important features in the treatment of children with DM. PMID- 12584539 TI - Hypovitaminosis D and vitamin D deficiency in exclusively breast-feeding infants and their mothers in summer: a justification for vitamin D supplementation of breast-feeding infants. AB - OBJECTIVE: To determine the prevalence of hypovitaminosis D in exclusively breast feeding infants and their mothers in a community where maternal sunshine exposure is low. STUDY DESIGN: Serum levels of calcium, phosphate, alkaline phosphatase, 25-hydroxy vitamin D (25-OHD), and intact parathyroid hormone were measured in 90 unsupplemented healthy term breast-feeding Arab/South Asian infants and their mothers in summer. Maternal dietary vitamin D intake was also estimated. RESULTS: The median age of infants was 6 weeks. The median serum 25-OHD concentrations in mothers (8.6 ng/mL) and infants (4.6 ng/mL) were low, and 61% of the mothers and 82% of the 78 infants tested had hypovitaminosis D (serum 25-OHD <10 ng/mL). The infants with hypovitaminosis D had elevated serum alkaline phosphatase and a tendency to higher serum intact parathyroid hormone levels. The average daily maternal vitamin D intake from commercial milk was 88 IU. CONCLUSIONS: Hypovitaminosis D is common in summer in exclusively breast-feeding infants and their mothers. The results provide justification for vitamin D supplementation of breast-feeding infants and mothers in the United Arab Emirates. Low vitamin D intake probably contributed to low maternal vitamin D status. PMID- 12584540 TI - Myeloid leukemia in Prader-Willi syndrome. AB - OBJECTIVE: To evaluate the frequency of cancers recorded by the Surveillance, Epidemiology, and End Results (SEER) Program in persons with Prader-Willi syndrome (PWS) METHODS: A survey was mailed in 1994 to 1852 registrants of the PWS Association (USA) inquiring about a diagnosis of any type of benign tumor or cancer (malignant tumor or leukemia). The risk of developing cancer was then estimated by comparing the observed number of cancers in the PWS population during 1975 to 1994 to the expected number in the general US population using data from the 1971-1994 SEER Cancer Statistics Review. RESULTS: Of the 1852 persons, 1160 (63%) responded, or 75% (1160/1552) of those who received the survey. The total number of observed cancer cases in the PWS study population was 8 versus 4.80 expected in the general US population (P =.1610). Three cases of myeloid leukemia were observed versus 0.075 leukemias expected (P =.0001). CONCLUSIONS: There appears to be an increased risk of myeloid leukemias, but not other cancers, among persons with PWS. PMID- 12584541 TI - Training of aerobic and anaerobic fitness in children with asthma. AB - OBJECTIVE: To assess the effect of a training protocol on aerobic and anaerobic fitness in children with asthma. STUDY DESIGN: Sixteen boys (mean age: 13 years; range: 10-16 years) with mild-to-moderate asthma participated in a rehabilitation program that included 6 weeks of individualized training on a cycle ergometer. Two groups were randomly formed: the control group (CG, n = 7) and the training group (TG, n = 9), which exercised at an intensity set at the heart rate corresponding to the ventilatory threshold, with 1-minute sprints against the maximal aerobic power (MAP) every 4 minutes. Session duration was 45 minutes, 3 sessions per week. Changes in maximal oxygen uptake (VO(2)max), MAP, short-term peak power (PP), and pulmonary function were assessed. RESULTS: Two patients of the training group did not complete the study. Pulmonary function remained unchanged in both groups. Improvement in both aerobic and anaerobic fitness was significant only in the training group (TG vs CG): VO(2)max +18% +/- 2.1% versus +9% +/- 4.5% (P <.05), MAP +32% +/- 5% versus 12% +/- 7% (P <.05), PP +21% +/- 5.7% versus +8.8% +/- 10% (P <.01). CONCLUSION: Exercise training with high intensity bouts is well tolerated in children with mild-to-moderate asthma. When included in a global rehabilitation program, this type of training improves both aerobic and anaerobic fitness. Anaerobic activities should be considered in sports rehabilitation programs for children with asthma. PMID- 12584542 TI - Hepatitis C virus infection associated with liver-kidney microsomal antibody type 1 (LKM1) autoantibodies in children. AB - OBJECTIVE: To evaluate the clinical pattern and evolution of chronic hepatitis C in children with liver/kidney microsomal antibody type 1 autoantibodies (LKM1). STUDY DESIGN: A multicenter, retrospective study, including the following groups of children with hepatitis C virus infection: (1). 21 consecutive LKM1-positive patients, (2). 42 age- and sex- matched LKM1-negative patients, and (3). 4 interferon-induced LKM1-positive cases. LKM1 reactivity to human microsomes and recombinant cytochrome P450IID6 (CYP2D6) was assayed by immunoblotting. RESULTS: Clinical and biochemical features overlapped in LKM1-positive and LKM1-negative children, but a fibrosis score >3 (range 0-6) was significantly more frequent (P =.04) in the former. Reactivity to microsomal protein and CYP2D6 was significantly (P =.02) associated with LKM1 titers >or=1:320 and was found in 39% of patients, including severe cases and both children (of 4 treated) who achieved a sustained alanine aminotransferase (ALT) normalization after steroid treatment. Five of 7 LKM1-positive children treated with interferon had an ALT exacerbation. CONCLUSIONS: LKM1-positive hepatitis C in children is characterized by a wide spectrum of biochemical, serologic, and histologic features. Whether autoimmunity may contribute to liver damage in a subgroup of patients with more severe liver disease, high LKM1 titers, and reactivity to CYP2D6 is a question deserving further investigation. PMID- 12584543 TI - A new mutation causing autosomal dominant periodic fever syndrome in a Danish family. AB - We describe four members in a family of 8 individuals over 3 generations with the autosomal dominant inherited periodic fever syndrome tumor necrosis factor receptor-associated periodic syndrome (TRAPS). The patients had recurrent episodes of fever, abdominal pain, arthritis, and rash. We examined the gene coding for the tumor necrosis factor receptor TNFRSF1A in all first-degree family members. In all 4 symptomatic members of the family, a hitherto undescribed mutation C98Y (380G-->A) in the TNFRSF1A gene was identified. In contrast, this mutation was not found in the 4 family members reported to be healthy nor in 50 normal control patients. The youngest member of the family, a 2-year-old boy, was treated successfully with etanercept. PMID- 12584544 TI - Disseminated cryptosporidium infection in an infant with hyper-IgM syndrome caused by CD40 deficiency. AB - We report the case of an infant with severe respiratory infections, chronic diarrhea, failure to thrive, and disseminated Cryptosporidium parvum infection. Laboratory investigations disclosed a diagnosis of hyper-IgM syndrome caused by CD40 deficiency. PMID- 12584545 TI - Surgical restoration of portal flow corrects procoagulant and anticoagulant deficiencies associated with extrahepatic portal vein thrombosis. AB - Extrahepatic portal vein thrombosis (EHPVT) is associated with abnormal circulating procoagulants and anticoagulants. Eleven children with EHPVT and abnormal coagulation factors underwent a mesenterico-left portal vein bypass to restore portal flow. Coagulation factors had returned to normal by one year. The data suggest that portal venous flow is essential for maintaining normal coagulation. PMID- 12584546 TI - Blood aspiration syndrome as a cause of respiratory distress in the newborn infant. AB - Early-onset respiratory distress and a radiographic appearance of an aspiration syndrome occurred in three neonates who had not passed meconium before delivery. In each case there was evidence of inhalation of blood, associated with very high plasma protein concentration in lung fluid. Blood aspiration syndrome is a distinct diagnostic entity that can result in significant respiratory distress in the neonate. PMID- 12584547 TI - The atopy patch tests for detection of cow's milk allergy with digestive symptoms. AB - Infants (n = 35) with digestive symptom were investigated for diagnosis of cow's milk allergy (CMA). Milk atopy patch tests (APTs) were positive in 19 of 24 CMA versus 1 of 11 in non-CMA patients (P <.001). This sensitivity (79%) and specificity (91%) suggest that the APT could improve the detection of conditions related to CMA. PMID- 12584548 TI - Hypercalciuria in Beckwith-Wiedemann syndrome. AB - We determined the incidence of hypercalciuria (HC) and its association with nephrocalcinosis and nephrolithiasis in 18 consecutive patients with Beckwith Weidemann syndrome (BWS). Random, nonfasting urine samples were obtained from each patient. All patients had abdominal ultrasonography, most on several occasions. Four patients (22%) had HC. Of these, 2 had nephrocalcinosis, one had hyperechoic kidneys, and one had normal renal imaging. Serum calcium was normal in all patients with HC. Because we found that an increased prevalence in the occurrence of HC and its complications in a group of children with BWS, any child with BWS should be evaluated for HC. PMID- 12584549 TI - A patient with type 2 Gaucher's disease with respiratory disease. AB - A 5-month-old boy had respiratory problems and gastroesophageal reflux. Electron microscopy of a tracheal biopsy specimen showed accumulation of lamellar bodies in the columnar cells indicative of lysosomal storage disease. Subsequently, the child had neurologic symptoms and hepatosplenomegaly, and the diagnosis of Gaucher's disease type 2 was made. PMID- 12584550 TI - Conjunctival mass: an unusual presentation of acute lymphoblastic leukemia relapse in childhood. PMID- 12584551 TI - How to avoid kernicterus. PMID- 12584552 TI - How to avoid kernicterus. PMID- 12584554 TI - The ambiguous role of nitric oxide in the developing brain. PMID- 12584556 TI - Chronic fatigue syndrome and Addison's disease. PMID- 12584558 TI - Nrf2 is an inhibitor of the Fas pathway as identified by Achilles' Heel Method, a new function-based approach to gene identification in human cells. AB - Here we describe the Achilles' Heel Method (AHM), a new function-based approach for identification of inhibitors of signaling pathways, optimized for human cells. The principle of AHM is the identification of 'sensitizing' cDNAs based on their decreased abundance following selection. As a proof of principle, we have employed AHM for the identification of Fas/CD95/APO-1 pathway inhibitors. HeLa cells were transfected with an antisense cDNA expression library in an episomal vector followed by selection with a suboptimal dose of the apoptotic inducer. Antisense inactivation of Fas inhibitors rendered the cells more sensitive to apoptosis resulting in their preferential death and consequent loss of their sensitizing episomes that were identified by subtraction. We show that the resulting products were enriched for sensitizing cDNAs as seven out of eight candidates tested were confirmed as inhibitors of Fas-induced killing either by transfection or by pharmacological inhibition. Furthermore, we demonstrate by multiple approaches that one candidate, NF-E2 related factor 2 (Nrf2), is an inhibitor of Fas-induced apoptosis. Inactivation of Nrf2 by antisense or by a membrane permeable dominant-negative polypeptide sensitized cells while overexpression of Nrf2 protected cells from Fas-induced apoptosis. In addition, dicumarol, an inhibitor of the phase II detoxifying enzyme NQO1, a downstream target of Nrf2, sensitized cells. Nrf2 induces the production of Glutathione (GSH) and we demonstrated that N-acetyl L-cysteine (NAC), a precursor to GSH, protected cells from Fas-mediated killing. Taken together, AHM is a powerful approach for the identification of inhibitors of a signaling pathway with a low rate of false positives that opens new avenues for function profiling of human genes and discovery of new drug targets. PMID- 12584559 TI - Role of TFG sequences outside the coiled-coil domain in TRK-T3 oncogenic activation. AB - The TRK-T3 oncoprotein, isolated from a human papillary thyroid tumor, arises from the fusion between the N-terminal domain of the TFG gene and the tyrosine kinase domain of the NTRK1 receptor. The 68 kDa TRK-T3 oncoprotein displays a constitutive tyrosine kinase activity resulting in its capability to transform NIH3T3 cells. The TFG portion of TRK-T3 contains a coiled-coil domain, which mediates protein oligomerization essential for the oncogene constitutive activation, and several consensus sites for protein interaction. In this study, we investigate the role of TFG sequences outside the coiled-coil domain on TRK-T3 activation, We constructed four mutants carrying different deletions of TFG sequences and expressed them in mammalian cells. By performing biochemical and biological assays we demonstrated that all the deleted regions are required for TRK-T3 activation, as they are involved in different mechanisms such as protein processing, formation of stable and/or functional complexes, and possible interaction with other proteins. By constructing site-specific mutants, we demonstrated a crucial role for a PB1 domain and a considerable contribution of an SH2-binding motif in TRK-T3 oncogenic activation. This work establishes an important role for TFG sequences outside the coiled-coil domain in the activation of the thyroid TRK-T3 oncogene. PMID- 12584560 TI - Interactions of the DNA mismatch repair proteins MLH1 and MSH2 with c-MYC and MAX. AB - MSH2 and MLH1 have a central role in correcting mismatches in DNA occurring during DNA replication and have been implicated in the engagement of apoptosis induced by a number of cytotoxic anticancer agents. The function of MLH1 is not clearly defined, although it is required for mismatch repair (MMR) and engagement of apoptosis after certain types of DNA damage. In order to identify other partners of MLH1 that may be involved in signalling MMR or apoptosis, we used human MLH1 in yeast two-hybrid screens of normal human breast and ovarian cDNA libraries. As well as known partners of MLH1 such as PMS1, MLH3 and MBD4, we identified the carboxy terminus of the human c-MYC proto-oncogene as an interacting sequence. We demonstrate, both in vitro by yeast two-hybrid and GST fusion pull-down experiments, as well as in vivo by coimmunoprecipitation from human tumour cell extracts, that MLH1 interacts with the c-MYC protein. We further demonstrate that the heterodimeric partner of c-MYC, MAX, interacts with a different MMR protein, MSH2, both in vitro and in vivo. Using an inducible c MYC-ER fusion gene, we show that elevated c-MYC expression leads to an increased HGPRT mutation rate of Rat1 cells and an increase in the number of frameshift mutants at the HGPRT locus. The effect on HGPRT mutation rate is small (2-3 fold), but is consistent with deregulated c-MYC expression partially inhibiting MMR activity. PMID- 12584561 TI - Decreased syndecan-2 expression correlates with trichostatin-A induced morphological changes and reduced tumorigenic activity in colon carcinoma cells. AB - The inhibition of histone deacetylase activity is known to induce morphological changes of transformed cells. In this study, we investigated the effect of the specific HDAC inhibitor, trichostatin A (TSA), on colon carcinoma cell lines. Treatment of human colorectal carcinoma cells, KM1214 and KM12SM, with TSA induced distinct morphological changes. Both cell lines, which normally piled up in layers without clear boundary, became more flattened, and formed monolayers with evident boundaries between cells, with concomitant increased actin filament organization. Cell-cell interaction was not affected much, based on expression level, membrane localization, and interaction of E-cadherin with beta-catenin. In contrast, syndecan-2 expression was dramatically reduced and it was correlated with the morphological changes of colon carcinoma cells. Consistently, downregulation of syndecan-2 expression by antisense cDNA clearly mimicked the morphological changes in KM12SM and reduced anchorage-independent growth of colon cancer cells. All these results indicate that reduced syndecan-2 expression correlates with TSA-induced morphological changes and reduced tumorigenic activity in colon carcinoma cells. PMID- 12584562 TI - Alpha6beta1 integrin induces proteasome-mediated cleavage of erbB2 in breast cancer cells. AB - ErbB2 and alpha6 integrin have been implicated in malignancy of breast cancer cells. Here we have determined the influence of alpha6beta1 integrin on erbB2 signaling in anchorage-independent growth, using MDA-MB435 breast cancer cells. Firstly, we transfected the cells with erbB2 cDNA, and isolated cells with high or low levels of alpha6beta1 integrin by cell sorting (alpha6H-ErbB and alpha6L ErbB). We found that an erbB ligand, heregulin beta1, enhanced growth activity of alpha6L-ErbB cells, but not alpha6H-ErbB cells. Secondly, we established cells expressing a beta4 integrin deletion mutant (beta4-deltacyt), which selectively inhibited alpha6beta1 integrin expression and adhesion to laminin-1. Again, heregulin beta1 enhanced the growth of erbB2 cDNA-transfected beta4-deltacyt cells, but not mock cells. Western blot analysis revealed that heregulin beta1 stimulated phosphorylation of Akt and its downstream molecules, GSK3beta and p70S6kinase, and that the extent of phosphorylation was greater in ErbB2/beta4 deltacyt cells than ErbB2/mock cells. Furthermore, we found that the erbB2 cytoplasmic domain was truncated in ErbB2/mock cells, which was independent of ligand stimulation and adhesion, and was suppressed by proteasome inhibitors. These results suggest that alpha6beta1 integrin inhibits erbB2 signals by inducing proteasome-dependent proteolytic cleavage of the erbB2 cytoplasmic domain, and may thereby contribute to the regulation of tumor growth. PMID- 12584563 TI - Screening for TP53 rearrangements in families with the Li-Fraumeni syndrome reveals a complete deletion of the TP53 gene. AB - The absence of detectable germline TP53 mutations in a fraction of families with Li-Fraumeni syndrome (LFS) has suggested the involvement of other genes, but this hypothesis remains controversial. The density of Alu repeats within the TP53 gene led us to search genomic rearrangements of TP53 in families without detectable TP53 mutation. To this aim, we adapted the quantitative multiplex PCR of short fluorescent fragments (QMPSF) method to the analysis of the 11 exons of TP53. We analysed 98 families, either fulfilling (six families) or partially meeting (92 families) the criteria for LFS, and in which classical methods had failed to reveal TP53 alterations. We identified, in a large family fulfilling the criteria for LFS, a complete heterozygous deletion of TP53. Additional QMPSF analyses indicated that this deletion, which partially removed the centromeric FLJ10385 locus, covered approximately 45 kb. This deletion was shown to result from a complex rearrangement involving two distinct Alu-mediated recombinations. We conclude that TP53 germline rearrangements occur as rare events, but must be considered in LFS families without detectable point TP53 mutation. PMID- 12584564 TI - Overexpression of KIT in chromophobe renal cell carcinoma. AB - We analysed gene-expression profiles in 15 surgical specimens of conventional, papillary, and chromophobe renal cell carcinomas (RCCs) using high-density oligonucleotide arrays. From about 12,000 genes targeted by the array, 67 were upregulated specifically in each histological type of RCC. The oncogene KIT was one of the genes whose expression was upregulated specifically in chromophobe RCCs. Immunohistochemical analysis demonstrated the KIT gene product on the cell membrane of chromophobe RCC in all cases, although it was not detected in conventional RCCs or non-neoplastic kidneys except for weak staining in the cytoplasm of renal tubules. These results suggest that each histological subtype of RCC has a unique gene-expression profile, and in particular indicates for the first time that KIT could be a useful marker for chromophobe RCC. As overexpression of KIT might be involved in tumor growth, KIT could be a new therapeutic target in this special type of RCC. PMID- 12584565 TI - Transgenic overexpression of IGF-II induces spontaneous lung tumors: a model for human lung adenocarcinoma. AB - Elevated levels of insulin-like growth factor (IGF)-II are associated with a poor prognosis in human pulmonary adenocarcinoma; however, a causal role for IGF-II in pulmonary adenocarcinoma has not been demonstrated. Here, we show that transgenic overexpression of IGF-II in lung epithelium induces lung tumors in 69% of mice older than 18 months of age. These tumors displayed morphological characteristics of human pulmonary adenocarcinoma such as their epithelial origin, tubulo-acinar architecture and expression of TTF-1, SP-B and proSP-C. Examination of signaling molecules downstream of the IGF-IR showed the activation of either the Erk1/Erk2 or p38 MAPK pathways, but not both, within the lung tumors. Notably, all lung tumors contained high levels of phosphorylated CREB, suggesting that both the Erk1/Erk2 and p38 MAPK pathways converged on this transcription factor. Moreover, IGF-II induced proliferation and CREB phosphorylation in human lung cancer cell lines, suggesting that IGF-II and CREB also contribute to the growth of human lung tumors. Thus, IGF-II is an important genetic factor in the development of lung tumorigenesis, in which activation of CREB is a ubiquitous event. The MMTV IGF-II transgenic mice provide a critical model for elucidating the role of IGF II in this fatal human disease. PMID- 12584566 TI - Essential role for the dimerization domain of NuMA-RARalpha in its oncogenic activities and localization to NuMA sites within the nucleus. AB - Nuclear mitotic apparatus protein-retinoic acid receptor alpha (NuMA-RARalpha) is the fourth of five fusion proteins identified in acute promyelocytic leukemia (APL) patients. The molecular basis for its oncogenic activity has not been delineated. In gel-shift assays, NuMA-RARalpha bound to retinoic acid response elements (RAREs) both as a homodimer and as a heterodimer with RXRalpha. The binding profile of NuMA-RARalpha to a panel of RAREs was very similar to PML RARalpha and PLZF-RARalpha. In transient transfection assays using HepG2 cells, NuMA-RARalpha inhibited wild-type RARalpha transcriptional activity, while it augmented STAT3 transcriptional activity. In GST-pull down experiments, NuMA RARalpha formed a complex with the corepressor SMRT, was released from the NuMA RARalpha/SMRT complexes by all-trans retinoic acid (ATRA) at 10(-7)-10(-6) M and became associated with the coactivator TRAM-1 at 10(-8) M ATRA. Studies comparing NuMA-RARalpha with NuMA-RARalpha(deltaCC) demonstrated that the dimerization or alpha-helical coiled-coil domain of NuMA was required for homodimer formation, transcriptional repression of wild-type RARalpha, transcriptional activation of STAT3, and stability of the NuMA-RARalpha/SMRT complex. Confocal fluorescent microscopy of HeLa cells was performed following transient expression of cyan fluorescent protein (CFP)-tagged proteins and incubation of cells with or without ATRA. Within the nucleus, CFP-NuMA-RARalpha exhibited a speckled pattern identical to that observed in cells transfected with CFP-NuMA. Furthermore, CFP NuMA-RARalpha colocalized with yellow fluorescent protein-tagged (YFP)-NuMA. In contrast, CFP-NuMA-RARalpha(deltaCC) exhibited a diffuse granular pattern within the nucleus, similar to RARalpha. These results indicate that the dimerization domain of NuMA-RARalpha is critical for each of the known oncogenic activities of NuMA fusion proteins as well as its sequestration to nuclear sites normally occupied by NuMA and is distinct from RARalpha. PMID- 12584567 TI - CCAAT/enhancer binding protein beta (C/EBPbeta)-2 transforms normal mammary epithelial cells and induces epithelial to mesenchymal transition in culture. AB - The transcription factor CCAAT/enhancer binding protein (C/EBP)beta is critical for normal growth and differentiation of the mammary gland. The intronless C/EBPbeta gene encodes a single mRNA that produces three protein isoforms, C/EBPbeta-1, -2, and -3, which share a common basic-leucine zipper domain at their C-terminus, but are distinguished at their N-termini by the in-frame methionine codon used to initiate translation. Although C/EBPbeta-1 and -2 are both transactivators, they likely perform distinct functions in mammary epithelial cells. C/EBPbeta-1 is the only isoform detected in normal human mammary tissue. In breast cancer cell lines, C/EBPbeta-1 is absent, and the C/EBPbeta-2 transactivator is expressed. Moreover, our data suggest that C/EBPbeta-2 is upregulated in human primary breast tumors. To assess C/EBPbeta 2's ability to participate in the transformation process, we generated recombinant retrovirus selectively encoding epitope-tagged C/EBPbeta-2. Strikingly, 10 days after infecting a normal human mammary epithelial cell line (MCF10A) with C/EBPbeta-2 virus, transformed subcultures were readily generated. Specifically, C/EBPbeta-2-overexpressing MCF10A cells form foci, gain anchorage independence, express markers associated with having undergone an epithelial-to mesenchymal transition, and acquire an invasive phenotype. These studies, and our previous observations, provide supportive evidence that deregulated expression of C/EBPbeta-2 contributes to malignant conversion of the human breast. PMID- 12584568 TI - Kinetics of v-src-induced epithelial-mesenchymal transition in developing glandular stomach. AB - The oncogene function in primary epithelial cells is largely unclear. Recombination organ cultures in combination with the stable and transient gene transfer techniques by retrovirus and electroporation, respectively, enable us to transfer oncogenes specifically into primary epithelial cells of the developing avian glandular stomach (proventriculus). In this system, the epithelium and mesenchyme are mutually dependent on each other for their growth and differentiation. We report here that either stable or transient expression of v src in the epithelium causes budding and migration of epithelial cells into mesenchyme. In response to the transient expression of v-Src or a constitutive active mutant of MEK, we observed immediate downregulation of the Sonic hedgehog gene and subsequent elimination of E-cadherine expression in migrating cells, suggesting the involvement of MAP kinase signaling pathway in these processes. v src-expressing cells that were retained in the epithelium underwent apoptosis (anoikis) and detached from the culture. Continuous expression of v-src by, for example, Rous sarcoma virus (RSV) was required for the epithelial cells to acquire the ability to express type I collagen and fibronectin genes (mesenchymal markers), and finally to establish the epithelial-mesenchymal transition. These observations would partly explain why RSV does not apparently cause carcinoma formation, but induces sarcomas exclusively. PMID- 12584569 TI - Delineating an oncostatin M-activated STAT3 signaling pathway that coordinates the expression of genes involved in cell cycle regulation and extracellular matrix deposition of MCF-7 cells. AB - A number of studies have demonstrated that the STAT pathway is an important signaling cascade utilized by the IL-6 cytokine family to regulate a variety of cell functions. However, the downstream target genes of STAT activation that mediate the cytokine-induced cellular responses are largely uncharacterized. The aims of the current study are to determine whether the STAT signaling pathway is critically involved in the oncostatin M (OM)-induced growth inhibition and morphological changes of MCF-7 cells and to identify STAT3-target genes that are utilized by OM to regulate cell growth and morphology. We show that expression of a dominant negative (DN) mutant of STAT3 in MCF-7 cells completely eliminated the antiproliferative activity of OM, whereas expression of DN STAT1 had no effect. The growth inhibition of breast cancer cells was achieved through a concerted action of OM on cell cycle components. We have identified four cell cycle regulators including c-myc, cyclin D1, c/EBPdelta, and p53 as downstream effectors of the OM-activated STAT3 signaling cascade. The expression of these genes is differentially regulated by OM in MCF-7 cells, but is unaffected by OM in MCF-7-dnStat3 stable clones. We also demonstrate that the OM-induced morphological changes are correlated with increased cell motility in a STAT3 dependent manner. Expression analysis of extracellular matrix (ECM) proteins leads to the identification of fibronectin as a novel OM-regulated ECM component. Our studies further reveal that STAT3 plays a key role in the robust induction of fibronectin expression by OM in MCF-7 and T47D cells. These new findings provide a molecular basis for the mechanistic understanding of the effects of OM on cell growth and migration. PMID- 12584570 TI - Inhibition of oxidative phosphorylation underlies the antiproliferative and proapoptotic effects of mofarotene (Ro 40-8757) in Burkitt's lymphoma cells. AB - In the search for retinoids active against Burkitt's lymphoma (BL), we found that the arotinoid mofarotene (Ro 40-8757) induced strong antiproliferative and apoptotic responses in most established BL cell lines as well as in primary BL cells. Ro 40-8757-induced apoptosis is associated with mitochondrial membrane depolarization, activation of caspase-3 and -9, and enhanced production of reactive oxygen species. These effects were related to a transient drop in intracellular ATP content, probably favored by a downregulation of NADH dehydrogenase subunit-1, a component of the mitochondrial respiratory chain (MRC) Complex I. Inhibition of MRC with thenoyltrifluoroacetone suppressed both the ATP recovery and apoptosis, confirming that the effects of Ro 40-8757 are mediated by changes in mitochondrial function. Compared to EBV-negative lines, EBV-carrying BLs were more resistant to Ro 40-8757-induced apoptosis. EBV infection and ectopic LMP-1 expression increased the resistance of BL cells to Ro 40-8757 induced apoptosis, probably through bcl-2 upregulation. Finally, we also show that 2-methoxyoestradiol, an inhibitor of the scavenger enzymes superoxide dismutases, enhanced Ro 40-8757-mediated apoptosis. These findings provide the rationale for evaluating the clinical efficacy of Ro 40-8757 in BL patients and suggest that the combination of Ro 40-8757 with inhibitors of scavenger enzymes may be a promising therapeutic approach for this aggressive lymphoma. PMID- 12584571 TI - Trans-activation of heparanase promoter by ETS transcription factors. AB - The remodeling of extracellular matrix (ECM) is an important process required for cancer cells to turn into invasive and metastatic cancer cells. To dissolve the protein components of ECM, matrix metalloproteinases are some of the essential enzymes. Another ECM remodeling enzyme is the heparanase (Hpa) that digests the heparin sulfate component of the matrix. In metastatic cancer cells the Hpa gene is upregulated. To investigate the mechanism of why Hpa was upregulated in metastatic cancer cells, the regulatory sequence of heparanase gene was isolated and its function analysed in metastatic breast cancer cells. We found there are four ETS transcription factor binding sites. Two of them flanking the transcription initiation of the Hpa gene are nonfunctional, whereas two others are highly functional and responded to exogenously added ETS transcription factors. Mutation of these two ETS binding sites abolished the transcriptional activation of Hpa promoter by ETS transcription factors. Among four transcription factors tested (ETS1, ETS2, PEA3, and ER81), ETS1 and ETS2 are more potent in transactivating the human Hpa gene. Furthermore, dominant-negative ETS transcription factors failed to transactivate Hpa promoter and could abrogate the function of wild-type transcription factor in transactivation activity of ETS transcription factors on the Hpa promoter. These results suggest that ETS transcription factors play an important role in tumor invasion and metastasis by modulating the remodeling of ECM. PMID- 12584572 TI - CpG island methylation in carcinoid and pancreatic endocrine tumors. AB - Carcinoid tumors and pancreatic endocrine tumors (PETs) are uncommon neuroendocrine neoplasms and their genetic alterations are not well characterized. CpG island methylation is a mechanism of gene silencing, and concordant methylation of multiple CpG islands as CpG island methylator phenotype (CIMP) has been described in tumors. The aim of this study was to evaluate CIMP in carcinoid tumors and PETs. We studied 16 carcinoid tumors, 11 PETs, and 22 associated normal mucosa or pancreas. Methylation status of the p14, p16, cyclo oxygenase 2 (COX2), O(6)-methyl-guanine methyltransferase (MGMT), estrogen receptor (ER), thrombospondin 1 (THBS1), retinoic acid receptor beta 2 (RARbeta), T-type calcium channel (CACNA1G), and multiple endocrine neoplasia type-1 (MEN1) genes, and of MINT1, MINT2, MINT25, MINT27 and MINT31 loci was evaluated by methylation-specific-PCR (MSP) or combined bisulfite restriction analysis (COBRA). Carcinoid tumors were frequently methylated at RARbeta, MGMT, p16, COX2, p14, THBS1, and ER ranging from 25 to 63% of tumors. Other CpG islands were infrequently methylated or unmethylated. The adjoining normal mucosa was also methylated for ER, COX2, and RARbeta, but methylation at p14, p16, THBS1, and MGMT was tumor-specific. By contrast, PETs and normal pancreas were frequently methylated only at ER. Methylation was more frequent in carcinoid tumors than PETs at MGMT (25 versus 0%, p = 0.03), THBS1 (44 versus 9%, p = 0.04), p14 (44 versus 9%, p = 0.04) and RARbeta (25 versus 0%, p = 0.03). Loss of p16 protein expression correlated with methylation of p16 gene in carcinoid tumors (p = 0.006). Our study indicates that methylation profile of carcinoid tumors differs from PETs, reflecting different molecular pathogenesis. PMID- 12584573 TI - The human caspase-2 gene: alternative promoters, pre-mRNA splicing and AUG usage direct isoform-specific expression. AB - Caspases have been shown to play important roles in apoptotic cell death, cytokine maturation and cell differentiation. However, the transcriptional regulation of the corresponding CASP genes remains poorly known. We describe a 5.1 kb fragment located upstream of the first translated exon in the human CASP-2 gene, which is known to encode caspase-2L and -2S protein isoforms. Transient transfection experiments, together with transcription start site mapping and transcript analysis, demonstrate that each caspase mRNA is initiated from separate promoter regions, and produced from alternative splicing events in these regions. The CASP-2L promoter is much stronger than the CASP-2S promoter, in good agreement with the respective transcript levels of the two caspases. In addition, several in-frame translational start sites can be identified for each isoform, one of which is common to both, present in the second common exon, and used efficiently. Surprisingly, the short isoform may also be initiated at a downstream AUG codon within the same exon. Thus, promoter strength, alternative transcriptional initiation and 5'-splicing events regulate the expression of the main caspase-2 isoforms that may be translated from alternative translation initiation codons. PMID- 12584574 TI - NORE1A, a homologue of RASSF1A tumour suppressor gene is inactivated in human cancers. AB - We recently demonstrated that RASSF1A, a new tumour-suppressor gene located at 3p21.3 is frequently inactivated by promoter region hypermethylation in a variety of human cancers including lung, breast, kidney and neuroblastoma. We have identified another member of the RASSF1 gene family by in silico sequence analysis using BLAST searches. NORE1 located at 1q32.1 exists in three isoforms (NORE1Aalpha, NORE1Abeta and NORE1B). Both NORE1A and NORE1B isoforms have separate CpG islands spanning their first exons. NORE1Aalpha Produces a 418 aa protein containing a Ras-association (RA) domain and a diacylglycerol (DAG) binding domain. NORE1Abeta produces a C-terminal truncation of the RA domain. NORE1B also contains the RA domain but not the DAG domain. NORE1 is the human homologue of the mouse Ras effector Nore1. No inactivating somatic mutations were found in lung tumour lines; however, NORE1A promoter region CpG island was hypermethylated in primary tumours and tumour cell lines. NORE1A promoter was methylated in 10/25 breast, 4/40 SCLC, 3/17 NSCLC, 1/6 colorectal and 3/9 kidney tumour cell lines, while NORE1B promoter was unmethylated in the same tumour cell lines. While 24% (6/25) of primary NSCLC underwent NORE1A methylation, methylation in SCLC was a rare event (0/22); (P = 0.0234). NORE1A expression in tumour cell lines was reactivated after treatment with a demethylating agent. There was no correlation between NORE1A and RASSF1A methylation status in NSCLC. Our results demonstrate that NORE1A is inactivated in a subset of human cancers by CpG island promoter hypermethylation, and in lung cancer this hypermethylation may be histological type specific. PMID- 12584575 TI - Nicotine patches and the subjective effects of cigarette smoking: a pilot study. AB - Twenty-five per cent of the North American population smoke cigarettes regularly. Twelve smokers (aged 19 to 55 years, Fagerstrom test score 3 to 10) participated in a double-blind, placebo controlled, counterbalanced study to determine the extent to which subjective effects of smoking are altered by nicotine delivered by transdermal patches. Subjects wore a placebo or 21 mg nicotine patch while abstaining from smoking for 48 h. Nicotine-mediated objective and subjective effects were measured at baseline, and after smoking a regular and a low yield cigarette at four different study sessions. Subjective effects were assessed using validated computerized questionnaires such as the Profile of Mood States and Visual Analogue Scales. Nonsmoking compliance was determined by measuring expired air carbon monoxide and saliva cotinine concentrations. Significant within-session differences were found in subjective effects at baseline and after smoking. No differences in subjective effects were found between patch treatments before smoking. Nicotine withdrawal symptoms such as craving, irritability, tension, frustration, anxiety and restlessness were significantly increased in both patch conditions after 48 h of smoking abstinence. Smoking markedly ameliorated nicotine withdrawal symptoms. Systolic blood pressure increased after smoking in both patch conditions (P=0.01). Visual Analogue Scale scores for cravings, nicotine effects, good effects and 'high' consistently increased after smoking during either patch condition. Thus, wearing a nicotine patch did not seem to modify the subjective effects of smoking compared with placebo in this preliminary study. PMID- 12584576 TI - Patterns of use and patients' attitudes toward complementary medications: a survey of adult general medicine patients at a major Canadian teaching hospital. AB - OBJECTIVE: To characterize the preadmission and hospital utilization of, and patients' attitudes toward, complementary medications. PATIENTS AND METHODS: Patient survey data were collected at an acute care hospital over a four-month period for 85 patients who had been admitted to the general medicine wards. Patient information about complementary medicine use and opinions were obtained through patient interviews. Patient demographic information including age, sex, ethnic background, level of education and annual household income were gathered. Patients were eligible to participate in the survey if they were on a general medicine ward on any given interview day. Patients were excluded if they were absent from their room, were sleeping, spoke no English and had no translator present, were occupied by other health care staff or friends and family on days of interviewing, or did not consent. Logistical regression was used to identify predictive factors for complementary medicine use. RESULTS: Eighty-five of 105 patients (81%) consented to complete the survey. The mean patient age was 55 years (range 19 to 91), and 45 respondents (53%) were male. About one-third of patients surveyed reported having previously visited a practitioner of natural medicine. Thirty-three patients (39%) interviewed indicated that they had used complementary medications at some time in their lives. One in six respondents (17%) reported using a complementary medicine immediately before admission to the hospital, although only two patients (3%) reported using a complementary medicine while in hospital. The most commonly reported products of use were echinacea, garlic pills and ginseng. Five patients (15%) indicated that their pharmacists were aware of their complementary medicine use, while 17 patients (52%) stated that their family doctors were aware. The majority of patients indicated they were satisfied with the information provided to them from various sources. Other data collected revealed that almost one-half would choose to take a complementary medicine for the ailment that brought them into the hospital. Univariate analysis indicated that age was a significant negative predictor for complementary medication use. CONCLUSIONS: The use of complementary medications on general medicine wards appears to be low; however, preadmission use of complementary medications appears common. The present study results suggest that use of complementary medications may go unreported; therefore, all health care providers should be aware of and knowledgeable about the potential self-administration of complementary medications by their patients. PMID- 12584577 TI - A basic conceptual and practical overview of interactions with highly prescribed drugs. AB - Drug interactions are frequently the result of altered activity of the mechanism(s) responsible for drug elimination. These include drug metabolism mediated by a select group of cytochrome P450 enzymes (CYP3A4, CYP2D6, CYP2C9, CYP2C19, CYP1A2) and drug transporters (P-glycoprotein). Adverse drug interactions can result from induction (loss of therapeutic benefit) or inhibition (increased toxicity from excessive effect) of drug elimination. CYPs and P-glycoprotein are discussed individually with regards to their characteristics, frequently prescribed drug substrates, inducers and inhibitors, and important adverse drug events. The potential for important drug interactions can be predicted based on the properties of the causative agent (oral bioavailability, mechanism of elimination, seriousness of adverse event) and the interacting agent. Consequently, drug interactions can be prevented by avoiding concomitant administration of interacting substances or possibly implementing alternative therapeutic strategies. Furthermore, susceptibility to adverse events depends not only on the interacting substances, but also on the patient and the method of drug administration. Commonly prescribed drugs that are unlikely to cause a drug interaction involving CYPs or P-glycoprotein are also discussed. PMID- 12584578 TI - Warfarin use in atrial fibrillation: A random sample survey of family physician beliefs and preferences. AB - BACKGROUND: In clinical practice, warfarin is underused for the prevention of stroke in individuals with atrial fibrillation despite unequivocal evidence of benefit and acceptable safety. OBJECTIVE: To ascertain, from primary care physicians, their beliefs and preferences regarding the use of warfarin in patients with atrial fibrillation. MATERIALS AND METHODS: A questionnaire was mailed to a random sample of 1000 primary care physicians in Ontario. Physician prescribing preferences from among treatment options available (warfarin, acetylsalicylic acid, ticlopidine, no therapy and other) were recorded for four separate scenarios of atrial fibrillation with varying degrees of risk for stroke. Physician perception of the risks associated with warfarin use and their awareness of the evidence of benefit were assessed. RESULTS: Three hundred twenty four physicians returned completed questionnaires. Among the four scenarios, physicians choosing not to use warfarin were three to six times more likely than physicians choosing to use warfarin to believe that there was inadequate evidence of benefit of warfarin for stroke prophylaxis, and they were four to six times more likely to be concerned about the risks of hemorrhage. These beliefs did not change significantly with scenarios describing patients with a high risk of stroke. CONCLUSIONS: Physician reluctance to use warfarin is associated with a false understanding of the benefit to risk ratio, which arises from a low appreciation of therapeutic benefits and a high concern regarding hemorrhage. PMID- 12584579 TI - A retrospective survey of the use of bupropion slow release by members of the Canadian Armed Forces. AB - BACKGROUND: Since the addition of bupropion slow release (Zyban, GlaxoSmithKline, Mississauga) to the Department of National Defence (DND) smoking cessation program (Butt Out), utilizing bupropion (Zyban) in combination with behaviour modification counselling, the Directorate of Medical policy has received several anecdotal reports from pharmacists and Canadian Forces members attributing significant side effects to the use of Zyban. As a result, the DND wished to assess the benefits versus the risks of using Zyban as part of the smoking cessation program. Subsequently, a retrospective review of the use of Zyban within the Canadian Forces over a one-year period was solicited to assess current policies. METHODS: Surveys were sent to Canadian Forces members receiving Zyban between September 1, 1998 and August 31, 1999. Members were questioned about smoking history and current status, perceived effectiveness of bupropion and both positive and negative experiences with the drug. Those reporting side effects and who had consented were contacted for an interview to obtain further details and information regarding the use of medical resources and effects on job performance. Members of the Canadian Forces visiting a doctor due to side effects were asked for permission to review their medical charts. RESULTS: Zyban was dispensed to approximately 1171 members over the one-year period and 357 responded to the survey. The point prevalence smoking cessation rate was 47% at the time of the survey and ex-smokers had been smoke-free for a mean of 181 days. Approximately 91% of ex-smokers and 52% of smokers found Zyban helpful in quitting. Side effects were reported by 252 members and 160 interviews were completed. Forty-three interviewees had seen a doctor because of side effects. Sixteen of the 43 charts were audited. Fifty-two respondents stated that side effects affected their ability to do their primary job. Two individuals were hospitalized for a total of six days. CONCLUSIONS: In light of the demonstrated effectiveness of Zyban and the overwhelming health benefits associated with smoking cessation, it is recommended that the current policies of funding for the DND smoking cessation program be left in place. The impact of Zyban's side effects on job performance and medical resources should be minimized through close monitoring and Zyban prescriptions should be dispensed in two-week quantities. PMID- 12584580 TI - The Toronto experience in diagnosing alcohol-related neurodevelopmental disorder: a unique profile of deficits and assets. AB - BACKGROUND: Fetal alcohol syndrome (FAS), which involves the triad of features reflecting facial dysmorphology, growth retardation and intellectual impairments, encompasses a relatively small proportion of the children affected prenatally by alcohol. Unfortunately, in the absence of facial dysmorphology, the diagnosis is difficult in the majority of children, who are considered to have alcohol-related neurodevelopmental disorder (ARND). Because accepted clinical methods are not pathognomonic, a novel profile approach was used to examine neuropsychological abilities and disabilities to identify children with ARND who do not meet the diagnostic criteria of FAS. OBJECTIVE: To establish a set of criteria, to be validated in future studies, for characterizing the neuropsychological profile of children with ARND. By describing the procedures at this preliminary stage of our work, the goal is to draw attention to this neglected topic and to suggest a model that can be replicated and validated by others, and to provide the first systematic clinical report on diagnosing ARND in Canada. PROCEDURES: On the basis of the literature, parents' descriptions and the authors' own experience with ARND, a profile of neuropsychological characteristics, including both deficits and assets, that are associated with prenatal alcohol exposure was hypothesized. A group of children was then evaluated, mostly adoptees or children in foster care, who were referred for learning and behavioural problems potentially associated with gestational exposure. Their results were submitted to a profile analysis by comparing their deficits and assets according to a list describing a hypothetical ARND profile to determine whether each child fit or did not fit the ARND profile. Groups were compared for background characteristics, FAS symptomatology, and results on specific neuropsychological and behavioural tests. Finally, the characteristics most strongly differentiating the two groups were identified. SETTING: Hospital-based outpatient program. PARTICIPANTS: Fifty-two children aged four to 18 years who were referred for a diagnostic assessment related to suspected or known prenatal alcohol exposure. OUTCOME MEASURES: Each child's assessment results were compared against a list of 21 deficits and six assets by two independent raters. Children with an average of more than 60% deficits and 50% assets were considered to have ARND, while the remainder were not. RESULTS: Twenty-eight children (54%) were assigned to the ARND group and 24 to the non-ARND group. The groups did not differ in physical features or home background characteristics, with the exception of higher parental intelligence quotient in the non-ARND group. The ARND group was more likely to have repeated a grade or received special education and scored lower on standardized measures of intelligence, language and memory abilities. Frequency of behaviour or social problems were equivalent in both groups. CONCLUSIONS: A profile approach used to identify children with ARND discriminates problems in neuropsychological but not behavioural domains. Because elevated scores on behavioural tasks in both ARND and non-ARND groups were clinically significant, more research is needed to identify what behavioural problems are unique to children with ARND compared with other clinic-referred children. PMID- 12584581 TI - JAK/STAT signaling pathways and cancer. Janus kinases/signal transducers and activators of transcription. AB - Recent highlights in understanding molecular nature of signaling pathways that mediate biological effects of various external stimuli and control number of normal physiological processes of cells such as growth, differentiation, senescence and apoptosis, defined three major groups of proteins which apparently play an essential role in transmitting external signals from surface membrane to target genes in the nucleus. These include Janus kinases (JAKs), signal transducers and activators of transcription (STATs) and their endogenous inhibitors of SOCS family. Their inappropriate functioning and defective cross talking associate with several human disorders including cancer. There is an increasing evidence that perturbances in STAT proteins are involved in the pathogenesis of some human malignancies. Moreover, cancer-related defective JAK/STAT/SOCS pathways may negatively affect tumor response to the cytokine-based immunotherapy. This article provides an overview of the current knowledge about JAK/STAT/SOCS intracellular signaling cascades with special emphasis on their abnormalities in cancer. PMID- 12584582 TI - The most frequent APC mutations among Slovak familial adenomatous polyposis patients. Adenomatous polyposis coli. AB - We screened 46 suspected families from whole Slovakia for familial adenomatous polyposis (FAP) cancer predisposition. Individuals were enrolled to the adenomatous polyposis coli (APC) gene mutations mapping program at the base of previous clinical investigation. We have used the following techniques: heteroduplex analysis (HDA), protein truncation test (PTT), single strand conformation polymorphism (SSCP) and sequencing for the identification and detailed positional analysis of APC mutations. Around 90% of all detected mutations were found being truncated. The most frequent mutations from this collection were located within codons 1309 and 1061 of exon 15 and represented 15% and 7%, respectively of all tested families. The expressive phenotype, large amount of colorectal polyps and congenital hypertrophy of the retinal pigment epithelium (CHRPE) were associated to all mutations within codons 1309 and 1060. PMID- 12584583 TI - Lipoxygenase inhibitors induce arrest of tumor cells in S-phase of the cell cycle. AB - Inhibitors of the lipoxygenase pathway of arachidonic acid metabolism represent a potential anti-tumor drugs. These compounds have been found to inhibit the growth and induce the apoptosis of various tumor cells both in vitro and in vivo. In this study, the effects of the lipoxygenase inhibitors esculetin and nordihydroguaiaretic acid (NDGA) on the progression of the cell cycle were investigated in eight mammalian cell lines of different origin. Flow cytometric analyses of cell cycle distribution after staining of DNA with propidium iodide or 7-aminoactinomycin D and DNA synthesis using incorporation of 5-bromo-2'-deoxy uridine showed that both esculetin and NDGA suppress cell growth by interrupting the progression of cells through S-phase that results in their accumulation in this phase of the cell cycle. The possible mechanisms of these effects and the significance of the findings for the improvement of anticancer therapy targeted on cell cycle is discussed. PMID- 12584584 TI - Is the central channel of nuclear pore complex really the exclusive gateway for macromolecular transport? AB - Examination of many nuclear pore complexes revealed in some of them very thin filaments presumably RNA with accompanying proteins, directing from inner nucleus to the edge of the complex. On one representative micrograph it is shown that this strand continues in the same direction through central part of the complex most probably through the peripheral channel. Next route of the strand is through the tunnel of a hollow rod - subunit of the cytoplasmic ring. PMID- 12584585 TI - A ratio of apoptosis to mitosis, proliferation pattern and prediction of radiotherapy response in cervical carcinoma. AB - The prognostic significance of apoptotic (AI) and mitotic (MI) indices, and the ratio of these parameters (AI/MI), MIB-1 labeling index (MIB-1LI) and proliferation pattern was studied in 130 (FIGO stage IB-IIIB) squamous cervical cancer patients before radiotherapy. Also the influence of the patients age and tumors pathological features (stage, grade, degree of keratinization) and DNA ploidy on the biological parameters were analysed. AI and MI were assessed on histological sections stained with hematoxylin and eosin, and the MIB-1LI on specimens stained with rabbit anti-human Ki-67 antibody (DAKO Ltd). Sections stained with MIB-1 antibody were used for assessment of the tumor proliferation pattern. The median age of the patients was 55 years (29-80). The median values for MIB-1LI, AI, MI, AI/MI, were: 52.3%, 1.1%, 1.5, and 0.9, respectively. In the univariate analysis median values for cut-off points were used for MIB-1LI, and AI, however, for other parameters significant cut-off points have been chosen. For MI it was 2.6 and for the AI/MI ratio 0.7. The median time of follow-up was 29 months, with a range of 2-145 months. The univariate analysis showed that tumor stage (p=0.7009), grade (p=0.6660) and AI (p=0.9378) had negligible influence on patients survival. However, MI >2.6 (p=0.0442), AI/MI 50, a significant difference in WBC count was shown in males (p=0.0590). The peripheral blood lymphocytes isolated from healthy subjects used as controls, exhibited undetectable, to low proportion of bcl-2 and p53 positive cells. Comparing the percentage of bcl-2 as well as p53 positive cells in controls and those of B-CLL patients, the significant difference for both proteins was observed (p=0.0009 and p=0.0001, respectively). The results of this study indicate that the overexpression of p53 protein may contribute to the downregulation of bcl-2 in a subgroup of our B-CLL patients. Considering the small numbers of tested p53 positive cases, it would be necessary to confirm our findings in a larger cohort of patients with longer follow up. Thus it would be possible to confirm our expectation of a possible value of the simultaneous aberrant expression of bcl-2 and p53 as useful predictors of future aggressive behavior of B-CLL. PMID- 12584587 TI - Embryotoxicity of cisplatin and a cisplatin-procaine complex (DPR) studied in chick embryo. AB - Cisplatin is widely used as an antitumor drug. To reduce its toxic side effects in patients, cisplatin has been bound with procaine in a cisplatin-procaine complex (DPR). The lethal and teratogenic effects of cisplatin alone and of complexed cisplatin were determined in the chick embryo in ovo in order to compare their influence on rapidly proliferating embryonic tissues. The embryotoxic (lethal + teratogenic) effect was examined after a single intra amniotic injection of one of six different doses, ranging from 0.03 to 30.0 microg, on embryonic days (ED) 3, 4 or 5. The minimal embryotoxic dose was lower for cisplatin alone (0.03-0.3 microg) than for cisplatin in the DPR complex (0.3 3.0 microg), suggesting that cisplatin alone is more embryotoxic than complexed cisplatin. Both substances caused malformations in the surviving embryos evaluated on ED 9. These malformations included microphthalmia, microcephaly, hypoplasia of the upper and lower jaw, cleft beak, and haemocephaly. Moreover, heart septum defects and limb reduction deformities were found after exposure to the DPR complex. The embryotoxicity of complexed cisplatin exhibited a stage response effect. It was highest on day 3 and gradually decreased until ED 5. Such an apparent stage-response effect was not observed for cisplatin alone. The embryotoxicity of procaine hydrochloride - a component of the complex - was also tested. Procaine hydrochloride alone did not produce any embryotoxic effect, not even after a single injection of the maximal tested dose (100.0 microg per embryo). We also examined the protective effect of procaine hydrochloride, whose separate administration at ED 4 was followed by the injection of 0.3 microg cisplatin. We did not observe any protective effect of procaine hydrochloride if injected separately. PMID- 12584588 TI - Activity of antioxidant enzymes and concentrations of thiobarbituric acid reactive substances (TBARS) in melanotic and amelanotic Bomirski melanoma tissues in the golden hamster (Mesocricetus auratus, Waterhouse). AB - The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSHPx), as well as the concentration of thiobarbituric acid reactive substances (TBARS) in tissues of transplantable melanoma in the golden hamster were measured and compared. Ten inbred male hamsters were used for the experiment. They were divided into two groups and were given Bomirski melanoma cells subcutaneously. The first group was given melanotic (Ma) melanoma cells. The second group was given amelanotic (Ab) melanoma cells. Thirty days after the transplantation the hamsters were dissected and the tumor tissues were taken and homogenized. A statistically significantly higher activity of the measured antioxidant enzymes was found in homogenates of Ma tumor than in homogenates of the Ab tumor. Activity of SOD is 8% higher in melanotic melanoma, 24% higher in CAT, and 45% higher in GSHPx. Statistically significant differences between TBARS concentrations were not confirmed. The higher activity of antioxidant enzymes in the melanotic tumor is a result of increased generation of oxygen-derived free radicals. It is presumed that it is strictly connected with intensified production of quinone and semiquinone radicals in the process of melanogenesis. PMID- 12584589 TI - The treatment of acute myeloid leukemia with mitoxantrone, etoposide and low-dose cytarabine in elderly patients - a report of Polish Acute Leukemia Group (PALG) phase II study. AB - The common dilemma in the treatment of elderly patients with acute myeloid leukemia (AML) is whether to use intensive myelosuppresive therapy with higher risk of treatment related mortality (TRM), but a chance for complete remission (CR), or to treat less intensively in order to prolong survival time with a better quality of life. The aim of this prospective, phase II study was to assess the efficacy and toxicity of low dose combination induction treatment consisted of cytarabine at a dose of 10 mg/m2 every 12 h s.c. for 7 days, VP-16 at a dose of 100 mg/day p.o. for 7 days and mitoxantrone at a dose of 6 mg/m2 i.v daily on days 1-3. Two induction courses were planned. In the group of 44 patients 12 (27%) achieved CR, 4 (9%) patients were in PR and there were 9 (20%) early deaths (ED). Age, performance status, preceding myelodysplastic syndrome, karyotype, WBC and % of blasts in bone marrow were not significant prognostic factors for CR probability. The following initial factors appeared to be related to a shorter duration of survival time from the start of treatment: age >70 (p<0.03), poor performance status (p<0.03), and % of BM blasts 50 (p<0.05). We conclude that, despite promising results in the pilot study the efficacy of this induction treatment is not better than the efficacy of other regimens. The hematological toxicity of this treatment seems to be comparable with "3+7" regimen. PMID- 12584590 TI - Quadruple cancer, including triple cancers in the head and neck region. AB - Multiple primary tumors are not rare: they are encountered in 3-5% of malignant tumors. They are particularly frequent in the head and neck [20]. They are most often met with secondary malignant tumors; triple tumors occur in only 0.5%, quadruple tumors in 0.3% of malignant tumors. The possibility of developing a second metachronous cancer 5 years after undergoing treatment of the initial head and neck cancer is approximately 22%. Multiple metachronous tumors often appear 3 4 years after the observation of the primary tumor, or even after 5-10 years in the case of laryngeal tumors. The frequency of multiple primary tumors in the head and neck region supports the "field cancerization" theory, according to which the inducing agents (primarily smoking and alcohol consumption) can initiate the tumorous degeneration at a number of sites in the oropharyngeal region. The authors report on a case in whom surgery for bladder tumor was followed 101 months later by tumor development in the region of the head and neck: 3 such tumors were treated within a period of 21 months. The histologic result on the bladder tumor was transitiocellular carcinoma, while the latter ones were squamous cell carcinomas. Three of the tumors were treated effectively (no local recurrence or metastasis developed), but the fourth led to the death of the patient. The literature on multiple tumors of the head and neck is reviewed, and possible etiologic factors are discussed. It is pointed out that, besides primary and secondary prevention, close observation of these patients is required, repeated panendoscopy of the upper aerodigestive tract and genetic examinations are recommended. PMID- 12584591 TI - Early predicting recurrent cervical cancer with combination of tissue polypeptide specific antigen (TPS) and squamous cell carcinoma antigen (SCC). AB - The establishment of new tumor marker combinations including strong lead-time effects in detecting recurrent cervical cancer appears to be warranted. This retrospective study includes 50 patients with recurrent squamous cell cervical cancer after operation or radiotherapy. The serial serum levels of the tumor markers tissue polypeptide specific antigen (TPS) and squamous cell carcinoma antigen (SCC) were determined. Cutoff values of 78.5 U/L for TPS and 1.5 microg/L for SCC were selected according to the 95th percentile of serum concentrations measured in healthy control patients. Comparing with other monitoring modalities, SCC and TPS showed lead-time effective in 27 and 30 cases, respectively. This difference was not statistically significant. The combination of SCC and TPS provided lead-time effects in 42 cases. Our data indicate that combination of TPS and SCC is a valuable tool in the early predicting recurrent cervical cancer. PMID- 12584592 TI - In vitro chemoresistance profile and expression/function of MDR associated proteins in resistant cell lines derived from CCRF-CEM, K562, A549 and MDA MB 231 parental cells. AB - Although cellular experiments have elucidated a number of active principles in the study of the multidrug resistance (MDR) phenomena, most of the drug resistant tumor cells were derived from different parental cell lines. This fact limits generalization of some experimental data and conclusions, and therefore we selected and characterized cell lines resistant to various anti-cancer agents derived from four parental cell lines: CEM (human T-lymphoblastic leukemia), K562 (human myeloid leukemia), A549 (human lung adenocarcinoma) and MDAMB 231 (human breast adenocarcinoma). In total we obtained a set of 42 resistant sublines, which is an excellent tool for the future studies of different aspects of MDR. In this study we report on some basic characteristics of these sublines, namely, cross-resistance to other anti-cancer drugs investigated by in vitro MTT assay, expression of MDR associated proteins (Pgp, MRP1, LRP, GST-pi and Topo IIalpha) as well as the functional activity of Pgp and MRP. PMID- 12584593 TI - Locoregional recurrence of breast cancer: a retrospective comparison of treatment methods. AB - The study was made to evaluate the clinical and pathological features of breast cancer patients with locally recurrent breast cancer and to assess the impact of the treatment method on their prognosis. Fifty-four patients with local recurrence after breast cancer were treated in Greatpoland Cancer Center between 1983 and 1995. It constituted 6.2% (54/878) of all patients with breast cancer treated in this period. Median length of interval between primary lesion and recurrence was 26.6 months, in 12/54 cases (22.2%) was longer then 5 years. Patients in time of recognizing primary breast cancer had tumor in clinical stage T2 (n=25) and T3 or T4 (n=29), in stage N0 (n=16) and N1 (n=36). Patients with recurrent breast cancer were treated using different methods. In 26 cases recurrent tumor was excited and then, in 15 cases irradiated, in 11 cases irradiated and additionally treated by chemotherapy or by hormonotherapy. In 28 cases patients were disqualified for excision due to local advance of disease. They were all irradiated and then treated by chemotherapy (n=17) or hormonotherapy (n=11). 5-year survival rates were compared with the chosen clinical factors (age, clinical stage, histopathology), length of interval between primary tumor and recurrence and with different methods of treatment including excision or not. 5-year overall survival rate was 33.3%. In locally advanced tumors (stage T3) the effect was worse then in stage T2 tumors. Five year survival rates after recurrence were 20.8% and 52.0%, respectively (p=0.001). No statistically important correlations between lymph node involvement, age, histology and survival rate were found. Differences between 5 year survival rate were observed according to length of interval between recognizing the primary lesion and recurrence. Patients with interval shorter then 24 months had survival rate 14.3%, between 24 and 60 months - survival rate 64.3% and with interval longer then 60 months - 41.7%. Statistically important differences were noted between first and second group (p=0.01) and first and third group (p=0.03). Patients treated with local excision followed by radiotherapy and/or systemic therapy had greater 5-year survival rate (53.9%) then patients disqualified for incision (14.3%) (p=0.0001). PMID- 12584599 TI - What is the value of telerobotic technology in gastrointestinal surgery? AB - BACKGROUND: Although telerobotic technology has entered clinical application, its value for gastrointestinal surgery is unclear. Our objective was to evaluate the performance characteristics of telerobotically assisted laparoscopic cholecystectomy (TALC). METHODS: All TALCs performed using the da Vinci Surgical System between January 2000 and September 2001 at a tertiary academic medical center were analyzed. RESULTS: For this study, 20 patients (80% female) with a mean age of 47 +/- 4 years underwent TALC. All had symptomatic cholelithiasis, and all had successful TALC results without complications or need for conversion to conventional laparoscopic cholecystectomy (CLP). The mean procedure time was 152 +/- 8 min. The procedures were performed by one of three staff surgeons experienced in laparoscopic surgery who had training in telerobotic surgery. The perceived advantages of TALC over CLP included easier tissue dissection, enhanced dexterity, and stimulated interest in biliary surgery. The disadvantages included increased operating time and lack of tactile feedback. CONCLUSIONS: The TALC procedure is effective and safe when performed by appropriately trained surgeons. Telerobotic technology has the potential to reinvigorate gastrointestinal surgery. PMID- 12584600 TI - Technical considerations in laparoscopic resection of gastric neoplasms. AB - BACKGROUND: The purpose of this study was to determine the use of different laparoscopic approaches in the management of gastric neoplasms based on tumor type and location. METHODS: We retrospectively reviewed the records of seven patients (3 men and 4 women) with 11 gastric lesions who were referred to our facility between March 2000 and October 2001 for laparoscopic excision of gastric neoplasms. RESULTS: Two patients had gastrointestinal stromal lesions (3 lesions); two patients had hyperplastic polyps (3 lesions); one patient had carcinoid tumor (2 lesions); one patient had a carcinoma in situ and an adenoma; and one patient had an ectopic pancreas. Extraluminal laparoscopic wedge resection was used in four patients with lesions at the anterior gastric wall or along the lesser or greater curvature. Intragastric excision was used in two patients with small posterior wall lesions, and a transgastrotomy approach was used in one patient with a posterior wall lesion that could not be removed by the intragastric approach. All the lesions were completely excised with clear margins. The median hospital stay was 3 days. Complications developed in two patients. One patient presented with a perforated ulcer 2 weeks after surgery, and a second patient had postoperative pyloric edema that resolved with conservative treatment. CONCLUSIONS: The use of different laparoscopic approaches based on gastric neoplasm type and location facilitates tumor access and resection. PMID- 12584601 TI - Sonographic evaluation for peripheral pulmonary nodules during video-assisted thoracoscopic surgery. AB - BACKGROUND: The clinical value of sonographic guidance during video-assisted thoracoscopic surgery (VATS) was studied in 26 patients with peripheral pulmonary nodules. METHODS: Twenty-six patients underwent VATS between June 2000 and March 2001 for primary lung cancer (n = 15), pulmonary metastasis (n = 6), and various benign tumors (n = 5). RESULTS: Sonographic guidance successfully visualized peripheral pulmonary tumors in 21 of the 26 patients (81%). Among them, rich intratumoral blood flow signal was identified in 8 of the 15 primary lung cancers by using color Doppler sonography. The peak velocity was 26 +/- 12.8 cm/s in primary lung cancer and 9.4 +/- 1.7 cm/s in pulmonary metastasis, respectively (p <0.01). CONCLUSION: Sonographic guidance during VATS is helpful for locating lesions and determining the extent of surgical resection. The color Doppler method was also useful for evaluating intratumoral blood flow, which yielded significant information for differentiating primary lung cancer, metastatic tumors, and various benign tumors. PMID- 12584602 TI - Can preconditioning reduce laparoscopy-induced tissue injury? AB - BACKGROUND: Pneumoperitoneum (P) created to facilitate laparoscopy (L) is associated with splanchnic perfusion, ischemia/reperfusion (I/R) injury, and oxidative stress. In this randomized controlled experimental study with blind outcome assessment, we evaluated the effect of preconditioning (PRE) on L-induced I/R injury. METHODS: The subjects were 40 Sprague-Dawley male rats. P was created in all except controls, using carbondioxide (CO2) insufflation under a pressure of 15 mmHg. PRE consisted of 10 min of P, followed by 10 min of deflation (D). The rats were randomized to the following groups: Group P was subjected to 60 min of P. Group P/D was subjected to 60 min of P, followed by 45 min of D. Group PRE/P was subjected to PRE, followed by 60 min of P. Group PRE/P/D was subjected to PRE, followed by 60 min of P and 45 min of D. Group C (control) was subjected to a sham operation, without P. Its anesthesia time was equal to that for group PRE/P/D. At the end of the experiments, the rats were killed; blood, liver, and kidney samples were then obtained and coded. Plasma alanine aminotransferase (ALT) and malondialdehyde (MDA), as well as homogenized tissue MDA levels and glutathione (GSH) activities, were measured; tissue samples were assessed for histopathological evidence of injury; all assessments were done by investigators blinded to the study design. The results were decoded and analyzed statistically with the Kruskal-Wallis and Mann Whitney tests. A p <0.05 was considered significant. RESULTS: Plasma ALT as well as plasma, liver, and kidney MDA levels and liver and kidney injury scores were increased, whereas liver and kidney GSH values were decreased in groups P and P/D, as compared to group C. Rats subjected to PRE before P had plasma ALT, kidney MDA, and kidney and liver GSH levels comparable to controls; their kidney and liver injury scores were higher than controls but significantly lower than nonpreconditioned animals. PRE enabled decreased plasma, kidney, and liver MDA as well as increased kidney GSH if applied before P; its efficacy on oxidative stress was limited to providing decreased kidney MDA and increased kidney GSH if applied before P/D. However, PRE significantly attenuated kidney and liver injury after P as well as P/D. CONCLUSION: PRE consisting of 10 min of P followed by 10 min of D decreases the oxidative stress induced by sustained P in the plasma, liver, and kidney. PRE significantly limits liver and kidney injury after prolonged P and P/D. After further studies to define its ideal timing, PRE before L incorporating P may have clinical relevance, especially for elderly patients or those with impaired hepatic and/or renal function or perfusion. PMID- 12584603 TI - Laparoscopic ileocecal resection in Crohn's disease: a case-matched comparison with open resection. AB - BACKGROUND: Despite some encouraging preliminary results, the role of laparosropic surgery in the treatment of Crohn's disease (CD) is a subject of controversy and still under evaluation. The aim of this case-matched study was to compare the postoperative course of laparoscopic and open ileocecal resection in patients with CD in order to define the potential role of laparoscopic surgery in CD. METHODS: From 1998 to 2001, 24 consecutive patients with isolated Crohn's terminal ileitis treated by laparoscopic ileocecal resection (laparoscopy group) were compared with 32 patients matched for age, gender, duration of disease, preoperative steroid treatment, fistulizing disease, and associated surgical procedure, and treated by open resection (open group). RESULTS: In the laparoscopy group, four procedures (17%) were converted. There were no deaths. The morbidity rate was 20% in the laparoscopy group and 10% in the open group (NS). There was no significant difference between the two groups in operating time, size of bowel resection and resection margin, postoperative morphine requirement, resumption of intestinal function, tolerance of solid diet, or length of hospital stay. CONCLUSIONS: Laparoscopic ileocecal resection in CD is safe and effective, even for fistulizing disease. There are no significant differences between laparoscopic and open ileocecal resection, especially in terms of the mortality and mortality rates. Consequently, because laparoscopic surgery seems to offer cosmetic advantages, it should be considered the procedure of choice for patients with ileocecal CD. PMID- 12584604 TI - Genomic characterization and expression of mouse prestin, the motor protein of outer hair cells. AB - We previously identified the gerbil gene (gPres) that encodes prestin, the putative motor protein responsible for outer hair cell (OHC) electromotility. Here we report the cloning and characterization of the complete genomic structure of the mouse Prestin (mPres) gene. We performed 5'- and 3'-RACE to determine the size and identity of the full-length mRNA transcript. The mPres gene encodes a protein 96% identical to the gPres gene product. The prestin open reading frames are 91% identical at the nucleotide level. Using an antibody raised against the N terminus of gerbil Prestin, we observed mPrestin expression by immunofluorescence in the lateral membrane of mouse OHCs and found no detectable expression elsewhere in the organ of Corti. On the basis of the available genomic sequence from mouse Chromosome (Chr) 5, we concluded that the mPres gene is centromerically related to and resides within 19 kb of, the Reln gene. We were also able to characterize the exon/intron junctions of mPres by using cDNA/genomic sequence comparisons, as well as exon-exon PCR and sequencing. The mPres gene has 18 exons that encode protein and two exons in the 5' UTR. A CpG island, located at the start of exon 1, is a potential transcription start site. Sequence analysis of the ~500 bp upstream from exon 1 revealed multiple potential transcription factor binding sites, including both TATA and GC boxes, as well as other regulatory-element binding sites. PMID- 12584605 TI - Hierarchical relationship between bone traits and mechanical properties in inbred mice. AB - Osteoporotic fracture incidence and underlying risk factors like low peak bone mass are heritable, but the genetic basis of osteoporosis remains poorly understood. Based on beam theory, stating that mechanical properties of a structure depend on both the amount and quality of the constituent materials, we investigated the relationship between whole bone mechanical properties and a set of morphological and compositional traits in femurs of eight inbred mouse strains. K-means cluster analysis revealed that individual femora could be classified reliably according to genotype based on the combination of bone area (tissue amount), moment of inertia (tissue distribution), and ash content (tissue quality). This trait combination explained 66-88% of the inter-strain variability in four whole-bone mechanical properties that describe all aspects of the failure process, including measures of brittleness. Stiffness and maximum load were functionally associated with cortical area, while measures of brittleness were associated with ash content. In contrast, work-to-failure was not directly related to a single trait but depended on a combination of trait magnitudes. From these findings, which were entirely consistent with established mechanical theory, we developed a hierarchical paradigm relating the mechanical properties that define bone fragility with readily measurable phenotypic traits that exhibit strong heritability. This paradigm will help guide the search for genes that underlie fracture susceptibility and osteoporosis. Moreover, because the traits we examined are measurable with non-invasive means, this approach may also prove directly applicable to osteoporosis risk assessment. PMID- 12584606 TI - Characterization and transcriptional analysis of the mouse Chromosome 16 cytokine receptor gene cluster. AB - The class II cytokine receptor (CIICR) genes Il10r2 and Ifnar1 are localized on mouse Chr 16 in a cluster that also contains the CIICR genes Ifnar2 and Ifngr2. The structure of the Il10r2 gene was deduced and consisted of 7 exons and 6 introns arrayed in an organization similar to its human ortholog. We also present a revised Il10r2 cDNA sequence with a total of 100 bp of additional nucleotide sequence in the 5' and 3' untranslated regions, and report the first extensive profiles of Il10r2 and Ifnar1 mRNA developmental stage and adult tissue expression. Promoter-luciferase reporter constructs were used to define the major region (-108 to +67) that conferred basal expression of the Il10r2 gene. Long range comparative genomic sequence analysis between the mouse and the orthologous human CIICR genomic loci revealed several conserved non-coding regions. The most proximal conserved non-coding sequence was a 204-bp element located 1.6 kb upstream of the transcriptional start site of Ifnar2 that had repressor-like activity in transient transfection assays with an SV40 promoter-luciferase reporter construct. The identification of multiple conserved non-coding sequences will provide the basis for further investigations to elucidate CIICR gene regulation. PMID- 12584607 TI - Exclusion of Madh2, Madh4, and Madh7 as candidates for the modifier of Min 2 (Mom2) locus. AB - We recently identified the Modifier of Min 2 (Mom2) locus. Mom2 is a new modifier of intestinal tumorigenesis that resulted from a spontaneous mutation in a B6 Apc(Min)/+ mouse. The presence of one resistant Mom2(R) allele results in a significant reduction in small intestinal polyp number and colon polyp incidence in Apc(Min)/+ mice. Through linkage analysis, we previously localized Mom2 to a 14-cM region on mouse Chromosome (Chr) 18, distal to the Apc gene. This region is syntenic with human Chr 18q, which frequently undergoes loss of heterozygosity (LOH) in several human cancers, including colorectal cancer. Residing in this region are the Madh2 and Madh4 genes, which have both been implicated in human colorectal cancer. Based on meiotic recombinations within the Mom2 region in the derivation of our congenic animals, we have narrowed the location of the Mom2 locus and excluded Madh2, Madh4, and Madh7, as well as Mbd1, Mbd2, Dcc, and Tcf4, as candidates for the Mom2 gene. PMID- 12584608 TI - Strain-specific modification of lethality in fucose-deficient mice. AB - The FX locus encodes an essential enzyme in the de novo pathway of GDP-fucose biosynthesis. Mice homozygous for a targeted mutation of the FX gene manifest a host of pleiotropic abnormalities including a lethal phenotype that is almost completely penetrant in heterozygous intercrosses on a mixed genetic background. Here we have investigated genetic suppression of FX-mediated lethality. Reduced recovery of heterozygous mice was observed while backcrossing the null FX allele to C57BL/6J (B6), but was less dramatic in an outcross to CASA/Rk and absent in an outcross to 129S1/SvImJ, indicating that genetic background modifies survival of FX+/- progeny. Substantial strain-specific differences in pre- and postnatal survival of FX-/- progeny were also detected in heterozygous crosses of C57BL/6J congenic, 129S1B6F1, and B6CASAF1 mice. Specifically, intrauterine survival of FX /- mice was greatly increased during a heterozygous intercross on a uniform C57BL/6J genetic background compared with survival on a hybrid genetic background consisting of a mixture of C57BL/6J and 129S2/SvPas. In addition, statistically significant clustering of FX-/- progeny into litters and specific breeding cages was noted during a B6CASAF1 FX+/- intercross, suggesting a rare mechanism for modifier gene action in which parentally expressed genes define the phenotype, in this case the survival potential, of mutant offspring. Our results disclose that lethality in FX mutant mice is determined by one or more strain-specific modifier loci. PMID- 12584609 TI - Mouse Sprr locus: a tandem array of coordinately regulated genes. AB - Small PRoline Rich (SPRR) proteins are primary constituents of the cornified cell envelope, necessary to create a permeability barrier across the body's surface. The family of murine Sprr genes has diversified, enabling the body to construct slightly different types of barriers as needed for backskin, mouth, tongue, etc. The Sprr genes have remained tandemly arrayed within 220 kb on mouse Chromosome (Chr) 3. On the basis of sequence similarity, we identified a novel member of the family, the murine ortholog of SPRR4. We present a sequence-verified physical map of the region and identify the complete coding sequence of the Sprr2 genes. Highly specific RNase protection assays based on the 3' untranslated sequences were used to query the expression of these genes in a model of barrier deficiency, mice with a targeted ablation of the transcription factor Kruppel like factor 4 (Klf4-/-). Twelve of the 15 members of the Sprr family are upregulated in the Klf4-/- mice. The sequences upstream of the start of transcription of the Sprr2 genes contain common regulatory elements conserved with the human SPRR2 genes. The clustering of the genes and their misregulation suggest that these genes may be held together in a tandem array to allow coordinate regulation. PMID- 12584610 TI - Comparative fine maps of bovine toll-like receptor 4 and toll-like receptor 2 regions. AB - Toll-like receptors are cell-surface receptors that activate innate and adaptive immune responses. We have used a 5000-rad, whole-genome radiation hybrid panel to map Toll-like receptor 4 (TLR4) to the distal end of bovine Chromosome (Chr) 8, and Toll-like receptor 2 (TLR2) to the proximal end of bovine Chr 17. To facilitate comparative mapping and contig construction, we have also used 5000- and 12,000-rad, whole-genome radiation hybrid panels to produce fine maps of the regions surrounding these genes in cattle. These fine maps triple the number of available markers in the TLR4 region and more than double the number of available markers in the TLR2 region. Comparative analyses show gene order conservation between the bovine Chr 8 region and human Chr 9, and between the bovine Chr 17 region and human Chr 4. In addition, the bovine Chr 8 region refines an evolutionary chromosomal breakpoint from a 10-megabase region to a 2.5-megabase region, and the bovine Chr 17 map suggests a new evolutionary chromosomal breakpoint. PMID- 12584611 TI - [Can that be really true?]. PMID- 12584612 TI - [Miniaturization and related trends in technology: not applicable for radiology?]. PMID- 12584613 TI - [Percutaneous radiofrequency ablation -- clinical indications and results]. AB - Percutaneous interstitial tumor ablation therapy is a relatively new and steadily growing field within interventional radiology. Radiofrequency ablation (RFA) became one of the most accepted techniques among hyperthermal ablation procedures, presumably due to its superior relation between probe diameter and efficacy. The first part of this review provided an overview of the underlying principles, technical background and application techniques. This second part describes the indications, techniques and results of clinical RFA applied to primary and secondary liver tumors as well as experiences with extrahepatic RFA applied to kidney, bone and lung, together with a discussion of the current literature. PMID- 12584614 TI - [Assessment of the quality of life of patients with peripheral vascular diseases]. AB - The Medical Outcome Short Form 36 is commonly used as a generic quality of life measure in the assessment of vascular disease. The Claudication Scale CLAU-S, the PAVK-86 Fragebogen, and the Walking Impairment Questionnaire WIQ are validated disease-specific questionnaires for intermittent claudication. A disease specific tool for critical ischaemia is lacking. Quality of life of patients with peripheral arterial occlusive disease is not only impaired in the physical functioning domains (mobility, self care, activities of daily living), but moreover in their social and emotional wellbeing. This situation worsens under conservative treatment. Angioplasty and operation produce similar improvements in all dimensions of hrQOL. As radiological interventional procedures just aim to improve hrQOL and do not bring a definite cure for the underlying disease, patients perception of their quality of life should be taken into account both in the indication for angioplasty and for the scientific evaluation of new treatment modalities. PMID- 12584615 TI - [Establishing a quality management system according to DIN EN ISO 9001:2000 in an academic radiological department]. AB - PURPOSE: Establishing a quality management (QM) system according the guidelines of DIN EN ISO 9001:2000 in an academic radiological department. MATERIALS AND METHODS: To fulfill the requirements of an academic radiological department a quality management system according the guidelines of DIN EN ISO 9001 : 2000 has been established within one year. All important educational, research and patient care issues have been discussed in plenary sessions of all employees of the department twice a week. All core processes of the department have been documented as process instructions and standard operation procedures. RESULTS: Supported by the staff unit "quality assurance" of the university hospital, the QM system has been established during one year. Assisted by all parties of the department, obliging regulations have been established which are well accepted in the daily routine clinical work but also in research and education. The implementation of the QM system caused an additional work load. However, spreading the work load by a consequent responsibility assignment, it could be reduced effectively. CONCLUSION: The implementation of a QM system within the daily routine work of an academic radiological department is feasible. It allows the installation of generally accepted rules which regulate the principal tasks of research and education, and patient care. PMID- 12584616 TI - [IHE (Integrating the Healthcare Enterprise): a new approach for the improvement of digital communication in healthcare]. AB - Parallel to the introduction of diagnosis related groups (DRGs) for the reimbursement of hospitals, a marked reduction of financial means within the healthcare system is taking place. Healthcare enterprise information systems will play an increasing role to accommodate the new working conditions by developing reliable and efficient workflow solutions. Interfacing the systems currently in use can meet considerable obstacles. By offering high connectivity, IHE (Integrating the Healthcare Enterprise), which was initiated by concerted actions of users and vendors, ensures improved health care delivery and, furthermore, assists in acquiring new information systems in the future. IHE is not a standard but makes extensive use of existing international standards, such as HL7 and DICOM. National IHE demonstrations confirmed the power of this approach and presented its mission to large groups of users and vendors. The concept continues to grow and for the first time provides groups of various interests cooperative solutions to the problems encountered in collecting and distributing information. PMID- 12584617 TI - [MR imaging of lung ventilation with aerosolized gadolinium-chelates]. AB - PURPOSE: To evaluate the feasibility of magnetic resonance assessment of human lung ventilation with aerosolized Gd-chelates in healthy volunteers. MATERIALS AND METHODS: Five healthy adults (mean age 37 years) were studied with a 1.5 T unit. The volunteers were instructed to inhale the aerosol through an airtight facial mask for 10 minutes. The aerosol was generated with a jet-type small particle nebulizer with attached heater. Ventilation imaging was performed using a respiration-gated dynamic T1-weighted turbo spin echo sequence (T(R) = 199 ms, T(E) = 8.5 ms, 12 signal averages, slice thickness 10 mm). Pulmonary signal intensity changes were calculated before and after nebulization. RESULTS: The investigation was successfully carried out in all volunteers. An acute or delayed allergic reaction to the aerosolized contrast medium was not observed. In 4 of 5 experiments (80 %), a homogeneous signal intensity increase was readily visualized with an average signal increase of 35 % after 10 minutes; in one experiment, the aerosol distribution was slightly heterogeneous. CONCLUSION: The results of the presented phase I clinical study demonstrate the feasibility of human ventilation imaging with aerosolized Gd-chelates for the first time. More trials with a larger number of healthy subjects and patients are needed before the clinical introduction of Gd-based ventilation MR imaging of the lungs. PMID- 12584618 TI - [Comparison of an edema-sensitive HASTE-TIRM sequence with delayed contrast enhancement in acute myocardial infarcts]. AB - GOAL: Comparison between a fluid-sensitive (HASTE-TIRM)-sequence and delayed contrast-enhancement in patients with acute myocardial infarct (AMI) in MRI. MATERIAL AND METHODS: 32 patients with AMI were imaged 7 +/- 4 days after the time of infarction with a 1.5 T unit using a T2 HASTE-TIRM and a contrast enhanced (CE) T1 turbo FLASH sequence. A threshold method (>2 SD in comparison with normal myocardium) was used to quantify the hyperintense zones in both sequences. The transmurality of the hyperintense regions was measured on a segmental basis. RESULTS: The hyperintense areas were larger on the HASTE-TIRM sequence with 29.6 +/- 13.2 % of the left ventricular (LV) area as compared to the CE-MRI with 19.2 +/- 10 % of the LV area (p < 0.0001). The measured transmurality was higher with the HASTE-TIRM sequence than with the CE-MRI (p < 0.0001). While the correlation between CE-MRI and peak creatine kinase (CK max) was good (r = 0.59, p < 0.001), no correlation was found between the HASTE-TIRM sequence and CK max (r = 0.29, p = ns). CONCLUSIONS: The peri-infarct edema can be depicted with a HASTE-TIRM sequence in addition to the non-viable infarct zone. The HASTE-TIRM sequence shows a higher transmurality of the hyperintense regions than the CE-MRI. The additional area depicted by the HASTE-TIRM sequence could represent functionally impaired but viable myocardium). PMID- 12584619 TI - [Prevalence and type of incidental extramammary findings in MRI of the breast]. AB - PURPOSE: To analyze prevalence and type of relevant incidental findings in patients undergoing breast MRI. MATERIALS AND METHODS: This prospective investigation consists of 1013 patients who underwent breast MRI as follow-up after breast cancer therapy, for pre-operative staging, and for screening of high risk patients as well as for clarification of unclear clinical examinations and inconclusive conventional mammography. Prevalence and type of relevant incidental extramammary findings were recorded together with the indication of the examination. RESULTS: Incidental extramammary findings were encountered in 92 (9%) of the 1013 patients. MRI had markedly more incidental extramammary findings with the staging examinations (39.5%) and follow-up examinations (11.6%). The prevalence of incidental malignant findings was 81% in patients examined for pre operative staging. Incidental benign and malignant findings were equally frequent in patients followed after breast cancer therapy. The incidental findings were exclusively benign in patients without a history of breast cancer. CONCLUSION: The interpretation of breast MRI should incorporate a careful analysis of the adjacent extramammary structures. Especially patients followed after breast cancer therapy can be expected to have incidental malignant findings outside the breast. PMID- 12584620 TI - [MRCP in primary sclerosing cholangitis]. AB - PURPOSE: Evaluation of MR-cholangiopancreaticography (MRCP) for the diagnosis of primary sclerosing cholangitis (PSC) in correlation with endoscopic retrograde cholangiopancreaticography (ERCP) and in comparison to the diagnostic accuracy of various T2-weighted sequences. METHODS AND MATERIALS: Fifty-five patients (34 males, 21 females; mean age 40 years, range 16 to 65 years) with suspected PSC were examined in a 1.5 T MR unit (Magnetom Vision, Siemens, Erlangen), using breath-hold transverse and coronal HASTE, paracoronal RARE and thin-sliced HASTE (TS-HASTE) sequences. Applying a five-point-scale, two blinded investigators assessed the image quality for ROC analysis. Morphologic criteria of PSC were documented and correlated with ERCP, which served as the gold standard, and sensitivity, specificity and diagnostic accuracy were calculated. RESULTS: PSC was confirmed in 40 of 55 patients (ERCP 55 of 55, liver biopsy 37 of 55), with concomitant chronic ulcerative colitis in 27 and Crohn's disease in 6 of the 40 patients. Qualitative analysis of the image quality showed no significant difference between RARE, HASTE and thin-sliced HASTE sequences (3.4/3.5/3.2). The RARE sequence had the highest sensitivity (97 %), specificity (64 %) and accuracy (84 %) for the detection of PSC. The difference between HASTE and thin-sliced HASTE was statistically significant (p < 0.01). Of the 40 patients with confirmed PSC, 29 were followed by MRI and 3 underwent a liver transplantation within the follow-up period. A Klatskin tumor, which was misdiagnosed by MRCP, was diagnosed by brush biopsy in 1 of the 40 patients. Interobserver variability was adequate to good (kappa 0.4 to 0.7), depending on the chosen sequence. CONCLUSION: MRCP seems to be a reliable non-invasive imaging method to diagnose and follow PSC. The RARE sequence showed the highest diagnostic accuracy of the T2-weighted sequences. PMID- 12584621 TI - [MR-urography and CT-urography: principles, examination techniques, applications]. AB - MR-urography (MRU) and CT-urography (CTU) provide refined imaging of the upper urinary tract not achievable with conventional intravenous urography (IVU). The traditional MR-urographic technique utilizes unenhanced, heavily T2 -weighted turbo spin-echo sequences for obtaining static fluid images of the urinary tract independent of the excretory renal function. T2 -weighted MR-urograms have proved to be excellent in visualizing the dilated urinary tract, even in non-excreting kidneys. In contrast, T1 -weighted MRU reflects the excretory renal function and displays the urine flow through the upper tract after renal excretion of an intravenously administered gadolinium chelate. The gadolinium-enhanced urine is visualized with fast T1 -weighted 3D-gradient-echo sequences. The combination of gadolinium and low-dose furosemide (5 - 10 mg) is the key for achieving a uniform distribution of gadolinium in the collecting system and for avoiding susceptibility artifacts (T2*-effects) in the urine. T1 -weighted excretory MRU provides impressive urograms of both non-dilated and obstructed collecting systems in patients with normal or moderately impaired renal function. Multislice CT-urography (MS-CTU) is also an excretory urography like T1 -weighted MRU. Furthermore, MS-CTU can be combined with low-dose furosemide for accelerated passage of excreted contrast material obviating the need for abdominal compression. CT-urography is limited by its radiation burden and the nephrotoxicity of radiographic contrast media. Combining MRU or MS-CTU with conventional MRI or CT offers several applications, e.g., diagnosis of intrinsic and extrinsic tumors. Meanwhile, MRU has replaced IVU in pediatric uroradiology and is also recommended for the assessment of renal transplants. MS-CTU may provide valuable information in chronic urolithiasis, especially if associated with a distorted urinary tract anatomy. Both MRU and MS-CTU will play an important role in modern uroradiology. PMID- 12584622 TI - [Sprengel deformity in a nine-year-old boy: diagnosis and therapy]. AB - Sprengel deformity is a congenital malformation with usually unilateral elevation and medial rotation of the scapula. An omovertebral bone is generally present. Associated skeletal malformations are frequently present. Though functional impairment is mild, the cosmetic and psycho-social impairment can be considerable. The prognosis of early surgical therapy is good. The presented case also justifies surgical correction in the older child. PMID- 12584623 TI - [Indirect MR-arthrography in the follow up of autologous osteochondral transplantation]. AB - PURPOSE: To evaluate the spectrum of findings in indirect MR-arthrography following autologous osteochondral transplantation. PATIENTS AND METHODS: 10 patients with autogenous osteochondral homografts underwent indirect MR arthrography at three, 6 and 12 months postoperatively. The MR protocol at 1.5 T comprised unenhanced imagings with PD- and T2 -weighted TSE- sequences with and without fat-suppression as well as T1 -weighted fat-suppressed SE-sequences before and after i.v. contrast administration and after active joint exercise. Image analysis was done by two radiologists in conference and comprised the evaluation of signal intensity (Sl) and integrity of the osseous plug and the cartilage surface, as well as the presence of joint effusion or bone marrow edema. RESULTS: At three months, all cases demonstrated a significant bone marrow edema at the recipient and donor site that corresponded to a significant enhancement after i.v. contrast administration. The interface between the transplant and the normal bone showed an increased Sl at three and 6 months in T2 -weighted images as well as in indirect MR-arthrography. The marrow signal normalized in most cases after 6 to 12 months, indicating vitality and healing of the transplanted osteochondral graft. The Sl of the interface decreased in the same period, demonstrating the stability of the homograft at the recipient site. The osteochondral plugs were well-seated in 9/10 cases. Indirect MR-arthrography was superior to unenhanced imaging in the assessment of the cartilage surface. Cartilage coverage was complete in every case. The transplanted hyaline cartilage as well as the original cartilage showed a significant increase of the Sl in indirect MR-arthrography, that did not change in follow up studies. There were no pathological alterations of signal and thickness alterations of the transplanted cartilage in follow up investigations. CONCLUSION: Indirect MR-arthrography is a useful diagnostic tool following autologous osteochondral transplantation. Assessment of transplant vitality, osseous fixation and stability is possible. PMID- 12584624 TI - [Radiation exposure of the head, midface and pelvis in multi-slice CT (MSCT): comparison with single-slice CT (SSCT)]. AB - PURPOSE: Comparison of the radiation exposure of the head, midface and pelvis in multi-slice CT (MSCT) and single-slice CT (SSCT) for manufacturer-implemented standard protocols. MATERIALS AND METHODS: An anthropomorphic Alderson-Rando phantom equipped with LiF-thermoluminescent dosimeters in the head, neck and pelvic region was examined with different MSCT and SSCT protocols, and the measured organ doses were compared. RESULTS: In comparison with the SSCT protocols, the MSCT protocols delivered about twice the organ dose to the head and pelvis and about eight times the organ dose to the midface. The comparison indicates that the parameters (slice collimation, mAs, pitch) used for the MSCT protocols deliver increased radiation doses. CONCLUSIONS: The current standard MSCT protocols generally lead to higher organ doses to the head, midface and pelvis than the SSCT protocols, raising the question whether using the MSCT protocols recommended by the manufacturer can be justified. The dose parameters should be optimized to achieve an acceptable balance of dose reduction and adequate image quality. PMID- 12584625 TI - [Transbrachial selective carotid DSA in outpatients: safety and efficacy]. AB - PURPOSE: To describe safety and efficacy of transbrachial selective carotid digital subtraction angiography (DSA) in outpatients. MATERIALS AND METHODS: From July 1999 to November 2001, selective carotid angiography was performed in 141 outpatients preferably using a left brachial arterial approach. The average age of the patients was 68 years (range: 39 - 89 years). After flush aortography through a 4F-pigtail catheter, bilateral selective common carotid artery (CCA) catheterization was performed with 4F-Sidewinder-1 or Sidewinder-2 catheters. In 49 patients, Doppler-sonography was performed before or after arteriography. A total of 41 patients underwent carotid surgery. RESULTS: Selective catheterization of the CCA was successful in 96 % of the cases. The diagnostic quality of the opacified aorta and CCA images was good to excellent. The overall complication rate did not exceed 2.1 %, and severe complications were not observed. One patient showed transient neurological symptoms and another mild nausea and agitation, probably as toxic reaction to the contrast medium. A single local hematoma developed after unsuccessful puncture of the brachial artery. CONCLUSION: Transbrachial selective carotid DSA is safe, reliable and well tolerated in outpatients and achieves a superimposition-free CCA visualization. PMID- 12584626 TI - [Endovascular brachytherapy to prevent restenosis after angioplasty]. AB - Endovascular radiotherapy is the first effective prophylaxis of restenosis after percutaneous transluminal angioplasty (PTA) and stenting. The FDA recently approved two devices for the delivery of intracoronary radiation following coronary artery stenting. Published multicenter, double-blind, randomized trials of intracoronary radiation therapy report good results for preventing in-stent restenosis, while the data for the peripheral circulation are still inconclusive. Beta-emitters are easier applicable and probably also safer, whereas gamma emitters have been more extensively evaluated clinically so far. Primary indication for endovascular brachytherapy are patients at high risk for restenosis, such as previous restenoses, in-stent hyperplasia, long stented segment, long PTA lesion, narrow residual vascular lumen and diabetes. Data from coronary circulation suggest a safety margin of at least 4 to 10 mm at both ends of the angioplastic segment to avoid edge restenosis. To prevent late thrombosis of the treated coronary segment, antiplatelet therapy with clopidogrel and aspirin are recommended for at least 6 months after PTA and for 12 months after a newly implanted stent. An established medication regimen after radiotherapy of peripheral arteries is still lacking. PMID- 12584627 TI - [Evolution of inflammatory parameters after fibroid embolization]. AB - PURPOSE: To describe the rise of classical laboratory tests for inflammation following transarterial uterine fibroid embolization (UFE) in order to monitor the normal course following UFE. MATERIALS AND METHODS: In 20 females, white blood cell (WBC) count and C-reactive protein (CRP) were determined before and up to 5 days after UFE. With the exception of one noninflammatory complication, the post-procedure course was uneventful in all patients. The measured values were correlated with both the total uterine volume and the amount of instilled embolizing agent. RESULTS: Following UFE, an increase in the WBC count to an average maximum of 10.8 +/- 3.5/wL (range 5.9 - 18.6/wL) was found. In 13 of 20 patients, the WBC count was above normal on at least one day following UFE. The increase reached the maximum on the third post-interventional day and subsided within 5 days after the UFE. The CRP values increased significantly to an average maximum of 41.9 +/- 28.8 mg/l. The maximum was found on the 2nd post interventional day in 8 patients, on the 3rd day in 11 patients and on the 4th day in one patient. No correlation to the total uterine volume or to the amount of the instilled embolizing agents was detected. CONCLUSION: Following uncomplicated UFE, a steep increase in CRP occurs with no or only a mild increase in the WBC count, which does not indicate an infected fibroid. The maximum is reached on the 3rd or 4th post-interventional day, followed by a decline in CRP and normalization of the WBC count. PMID- 12584628 TI - [Transjugular liver biopsy: trucut or aspiration biopsy with modified Ross needles?]. AB - PURPOSE: The transjugular liver biopsy is performed in patients with coagulopathy to evaluate diffuse hepatopathies. Using a jugular venous access, a liver vein is cannulated and a transvascular biopsy taken. The two techniques in use are the aspiration biopsy with a modified 15G or 16G Ross needle used for TIPS and the core biopsy with a Trucut needle. The sampling is quite similar for these techniques, but the costs are significantly different. This study is a meta analysis of data published on both techniques to search for clinical evidence that justifies the use of the more expensive Trucut needle or that finds aspiration with the less expensive Ross needle equivalent. MATERIALS AND METHODS: The study includes 16 publications that were published between 1996 and 2002 and provided reliable information on technical success rate, complications and adequate histology. The data on 1374 procedures were analyzed, comprising 436 aspiration biopsies and 938 Trucut biopsies. RESULTS: Considering both procedures together, 95.9 % of the procedures were technically successful and 92.2% of the histologic samples diagnostically adequate. The technical success rate was 96.7 % for the Trucut-biopsies and 94.4 % for the aspiration biopsies. The corresponding rates of obtaining diagnostically adequate histologic samples were 92.5 % and 91.5 %, respectively. The complication rate of the entire population studied was 0.65 % without significant difference found between the two methods. With a list price of 420 Euro, the Trucut system costs about 200 Euro more than the Ross needle, which is listed as approximately 220 Euro. CONCLUSION: In the absence of any significant differences between both techniques for transjugular liver biopsies, economic considerations make the aspiration biopsy with the modified Ross needle preferable over the Trucut needle biopsy. PMID- 12584629 TI - [A novel hirudin coating of vascular endoprostheses: experimental results]. AB - PURPOSE: Does hirudin coating improve the patency of iliac artery endoprostheses in comparison to non-hirudin-coated endoprostheses? MATERIALS AND METHODS: Nitinol stents and stentgrafts covered with polytetrafluoroethylene (PTFE) were coated with the polymer polyamino-p-xylylene-co-poly-p-xylylene using chemical vapor deposition (CVD) technique. Hirudin was covalently bound to the surface of the endoprostheses via the amino-group. External factors (mounting of the prosthesis, sterilization, storage time and temperature, release) affecting the hirudin activity were evaluated in vitro. Five types of prostheses were compared in vivo: (1) plain and (2) CVD- and hirudin-coated stents; (3) plain, (4) CVD coated, and (5) CVD- and hirudin-coated PTFE-stentgrafts. In 20 sheep, 16 protheses of each type were inserted in arteries pretreated with a Fogarty maneuver. The animals were followed for either 1 (n = 10) or 6 (n = 10) months. Immediately after implantation and after 1, 3, and 6 months, intravascular ultrasound (IVUS) and angiography were performed. The vascular specimens were analyzed histologically. RESULTS: Within 10 weeks, the hirudin activity of coated stents dropped 60 % due to external factors; the activity of coated PTFE stentgrafts dropped 20 %. After 1, 3, and 6 months, IVUS and histology revealed a significantly reduced patency of the hirudin-coated stentgrafts compared to the other prostheses. Only IVUS showed a significantly reduced patency of hirudin coated stents after 1 and 3 months compared to plain and CVD-coated PTFE stentgrafts. The reduced patency was caused by neointimal hyperplasia. CONCLUSIONS: In an experimental setting, hirudin coating did not improve the patency of vascular endoprostheses. PMID- 12584630 TI - [MR-guided coil embolisation of renal arteries in an animal model]. AB - PURPOSE: To test the feasibility of MR-guided coil embolization with passive visualization in an animal model. MATERIALS AND METHODS: All experiments were performed on a 1.5 T interventional MR-scanner (Gyroscan ACS-NT, Philips, Netherlands). A gradient echo sequence was combined with radial k-space acquisition and sliding window reconstruction technique to calculate the raw data with a specially designed back projector, yielding a frame rate of 20 images per second. The images were directly displayed on LCD-screens next to the MR scanner. A multipurpose catheter with dysprosium markers was placed into the renal arteries of two pigs under MR guidance, and a nitinol coil and platinum coil each were deployed. The position of the coils was verified with magnetic resonance angiography and conventional angiography. RESULTS: MR-guided catheterization of renal arteries with subsequent coil embolization was successfully carried out but the configuration of the coils could not be visualized on the MR images. CONCLUSION: MR-guided coil embolization of renal arteries is feasible. Exact guidance of coil deployment is not attainable with passive visualization. PMID- 12584631 TI - [Interstitial T1-weighted MR lymph fistulography with Gadomer-17 in an experimental animal model]. AB - PURPOSE: To visualize and localize fistulas of the thoracic duct with interstitial T 1 -weighted MR lymphography using Gadomer-17. MATERIALS AND METHODS: In 10 domestic pigs, leaks of the thoracic duct were created surgically or interventional-radiologically. The lymphatic leakage was located within the abdominal portion of the thoracic duct in 5 pigs, within the thoracic portion of the thoracic duct in 3 pigs, and in both, abdominal and thoracic portions of the thoracic duct, in 2 pigs. Subsequently, 10 micro mol/kg KG Gadomer-17 (1.5-1.8 ml) was administered interstitially in both hind legs of the animals. MR lymphography was performed with a 1.5 T MR unit using two different 3D gradient echo sequences before and 10 - 90 minutes after administration of contrast material. RESULTS: Leaks within the abdominal portion of the thoracic duct were directly visible as opacified fistulas. Indirect signs of active lymphatic fistulas were increasing extravasations of contrast material and free abdominal fluid. The 3D gradient echo sequence with the highest planar resolution (TR = 8,7 - 8,8 ms, TE = 4,2 - 4,3 ms, FA = 40 degrees, matrix size = 327 x 512) was best suited for distinct delineation of the lymphatic system and detailed demonstration of the thoracic duct fistulas. Intrathoracic leaks could not be demonstrated by MR lymphography due to reduced lymphatic flow or extravasated contrast medium at the abdominal puncture site. CONCLUSION: Interstitial MR lymphography with Gadomer-17 allows sensitive detection and localization of abdominally located leaks of the thoracic duct. PMID- 12584632 TI - Vascular capsule for telemetric monitoring of blood pressure. AB - PURPOSE: Development and experimental evaluation of an intravascular monitoring system for telemetric measurement of blood pressure and heart rate. MATERIALS AND METHODS: The monitoring system consists of an implantable silicone capsule (diameter 2.3 mm), containing a dedicated microchip with pressure sensors and signal-processing circuits as well as an antenna for wireless data and energy transfer using 6.78 MHz transponder technology. Three self-expanding legs at one end of the capsule served as a mechanism to lock the capsule at an arterial branch. A flow model, driven by a ventricular assist system, was used for testing and optimizing the implantation equipment, for checking the anchoring mechanism and for ensuring transmission of the measured pressure to the readout unit. In vivo experiments were performed in 8 minipigs (weight 25 to 30 kg), with three capsules placed in each minipig via the femoral artery using a dedicated 8-F sheath/pusher system. Follow-up was by CT angiography for up to 6 months after implantation. RESULTS: Flow model tests revealed a maximum deviation of pressure and heart rate measurements of 5% from the reference measurements. Signal transmission was reliable over a distance of 3 to 4 cm. Fluoroscopically guided in-vivo implantation of the capsules was simple and straightforward. In arteries with a diameter of 5 to 6 mm, the capsules were permanently fixed with one or two legs interlocked in side branches and without occlusion within 6 months. Three capsules developed a small non-occlusive appositional thrombus attached to the downstream (leg) part of the capsule. CONCLUSION: Our in-vitro and in-vivo experiments demonstrate the feasibility of wireless transmission from a capsule with a sufficient resolution of the sensor output signals to determine blood pressure and pulse rate. As long as the vessel diameter is wide enough, arterial fixation of the capsule does not induce thrombotic occlusion of the parent artery. With respect to future clinical applications, further refinements of the transmission technology are needed to extend the transmission distance between capsule and reader antenna. The technology of intelligent implants has further implications, such as monitoring of other physiological parameters, and the design of a control loop, which may be used for therapeutic feedback. PMID- 12584633 TI - [Schwannoma of the geniculate ganglion]. PMID- 12584634 TI - [Recanalization of an a. lusoria occlusion with PTA and stent]. PMID- 12584635 TI - [Wolf in sheep's clothing--bronchioloalveolar carcinoma]. PMID- 12584636 TI - Standardized reprocessing of reusable colonoscopy biopsy forceps is effective: results of a German multicenter study. AB - BACKGROUND AND STUDY AIMS: National and international guidelines recommend that a standardized protocol consisting of cleaning, ultrasound cleaning, and sterilization should be used for the reprocessing of endoscopic accessories in order to reduce the risk of transmission of microorganisms. This German multicenter study investigated the efficacy of standardized reprocessing of reusable biopsy forceps used during colonoscopy. MATERIALS AND METHODS: Ten endoscopy centers (eight hospitals and two private practices) used 330 biopsy forceps during routine colonoscopy. The forceps were used once, five times, or 20 times for colonoscopy, based on a randomization plan. The reprocessing protocol consisted of manual cleaning with an enzymatic agent, ultrasound cleaning with an enzymatic agent (30 min, 40 degrees C, 47 Hz), neutralization, drying, and sterilization (5 min, 134 degrees C). Aldehydes were not used, and the protocol did not include a disinfection step. The biopsy forceps were sent to three microbiological institutes, based on a randomization plan, to have them tested for the presence of organisms, including identification of bacteria. RESULTS: A total of 318 of the 330 forceps were evaluable; 314 forceps (98.74 %) were sterile after use once, five times, or 20 times. Four forceps were contaminated with Staphylococcus epidermidis (n = 2), Bacillus licheniformis (n = 1) and Corynebacterium aquaticum (n = 2). All of 25 forceps were sterile after being used 20 times. CONCLUSION: Colonoscopy biopsy forceps can be reliably reprocessed following this standardized protocol, even without aldehydes. PMID- 12584637 TI - Endoscopic mucosal resection in the management of gastric carcinoid tumors. AB - BACKGROUND AND STUDY AIMS: Gastric carcinoid tumors are a rare disease. Previously, total gastrectomy was regarded as the treatment of choice. However, differences in biological malignancy have recently led to the increased use of endoscopic mucosal resection (EMR) for treatment. We studied the outcome of EMR in patients with gastric carcinoids who were treated at our hospital and discuss the indications for endoscopic treatment. PATIENTS AND METHODS: Between 1986 and 1999 we carried out gastric mucosal resection in five patients with gastric carcinoid tumors. The procedure used for EMR was either strip biopsy or endoscopic aspiration mucosectomy. RESULTS: The carcinoid tumors measured 10 mm or less in four of the five patients. Two patients had type A gastritis, and all had hypergastrinemia. There was no evidence of recurrence during follow-up (range 6 - 66 months; mean 32.6 months). CONCLUSION: EMR is useful in the management of type 1 gastric carcinoids as classified by Rindi (hypergastrinemia; tumor diameter of 10 mm or less). PMID- 12584638 TI - Palliation of esophageal carcinoma with a new self-expanding plastic stent. AB - BACKGROUND AND STUDY AIMS: Self-expandable metallic covered stents (SEMS) are widely used for the palliation of esophageal cancer. In the long term, a drawback of these stents is the occurrence of nontumoral occlusion at their proximal or distal parts. New self-expanding plastic stents (SEPS) may combine the advantages of SEMS with those of previously used plastic stents. Our aim was to study prospectively the implantation of such stents in a series of patients with fairly long life expectancy, focusing on safety, feasibility and long-term effectiveness. PATIENTS AND METHODS: Between June 1999 and December 2000, in a prospective cohort study, 33 patients were treated with Polyflex stents for palliation of esophageal stenoses, and were followed up until death. The inclusion criteria demanded the presence of a nonresectable or nonoperable, histologically proven, malignant stricture of the esophagus causing significant dysphagia. RESULTS: Stent insertion was successful in all cases. The mean duration of follow-up of the patients was 149.7 days (range 25 - 469). A complete follow-up until death was documented for all patients (n = 33). During the entire follow-up, no stent occlusion was observed which was caused by either nontumoral or tumoral ingrowth, or by nontumoral overgrowth. Stent occlusions were caused only by tumoral overgrowth, and occurred at a rate of 12.1 % (n = 4). The stent migration rate was 6.0 % (n = 2), and the re-intervention rate overall was 21.1 % (n = 7). CONCLUSIONS: These results show that SEPS are effective for the palliation of dysphagia in patients with esophageal malignancies. The immediate results are similar to those observed with SEMS, and the observed low rate of late obstruction suggests that their long-term efficacy might be superior to that of SEMS. This device warrants evaluation in a controlled prospective trial. PMID- 12584639 TI - Argon plasma coagulation after piecemeal polypectomy of sessile colorectal adenomas: long-term follow-up study. AB - BACKGROUND AND STUDY AIMS: The aim of this study was to evaluate prospectively the long-term outcomes of using argon plasma coagulation (APC) as an adjunct to piecemeal polypectomy of large sessile colorectal adenomas. PATIENTS AND METHODS: A total of 77 patients with 82 sessile colorectal adenomas (median size 2.9 cm, range 1.5 - 8.0 cm) underwent snare piecemeal polypectomy. Patients in whom polypectomy was complete received no further treatment (polypectomy group; n = 14). When polypectomy was incomplete, additional treatment with APC was started either immediately or 1 - 3 months after the last polypectomy session (polypectomy + APC group; n = 63). Patients were followed (by endoscopy and biopsy) at regular intervals. RESULTS: Histologically proven adenoma eradication was achieved in 100 % of patients (14/14) in the polypectomy group and in 90 % of patients (57/63) in the polypectomy + APC group (two patients died of unrelated causes before adenoma was eradicated, two underwent operation because cancer was detected in the polyp treated, and two underwent operation because of endoscopic treatment failure). The adenoma recurrence rate was 14 % in both the polypectomy and polypectomy + APC groups. All recurrences except one occurred during the first year of follow-up and all were successfully re-treated endoscopically. A total of 69 patients in whom long-term follow-up data are available are free from adenoma at a median follow-up of 37 months (range 12 - 80). No major complications of endoscopic treatment occurred. In seven cases (9 %) the polyp was eventually shown to be malignant; in two of these patients the diagnosis of cancer was delayed as a result of unsuccessful endoscopic treatment. CONCLUSIONS: APC used in combination with piecemeal polypectomy of large colorectal adenomas is an effective and safe method of therapy, provided patient selection is careful and follow-up close. PMID- 12584640 TI - Endoscopic ultrasound-guided fine-needle aspiration of melanoma metastatic to the pancreas: report of two cases and review. AB - Malignant melanoma is a neoplasm which originates from melanocytes. Surgical exploration for abdominal metastases of cutaneous melanoma reveals pancreatic metastases in 16 % of patients which is usually accompanied by other intra abdominal spread. Endoscopic ultrasound (EUS) and EUS-guided fine-needle aspiration (EUS-FNA) of primary and metastatic pancreatic masses have already been described. We report the first use of EUS-FNA for the diagnosis of malignant melanoma metastatic to the pancreas. As our cases illustrate, these lesions may present as either cystic or solid pancreatic masses. Preparation of a cell block from the FNA aspirate and use of immunostaining may help to obtain the diagnosis. Despite thorough examination of the skin and mucous membranes, however, a primary site might not be found in these patients. PMID- 12584641 TI - Spontaneous passage of a dislocated esophageal metal stent: report of two cases. AB - Delayed transpyloric impaction is a very rare complication of esophageal metallic stent placement. Authors report different endoscopic removal techniques, but none of these has been validated yet because of their variable success, time-consuming nature, and risk of perforation and hemorrhage. We report that a "wait-and-see" approach is a safe and effective policy in patients with permeable transpyloric impacted esophageal stents. PMID- 12584642 TI - Endoscopically assisted video capsule endoscopy of the small bowel in patients with functional gastric outlet obstruction. AB - Since the introduction in 2001 of M2A video capsule imaging of the small bowel in humans, this technique has been used increasingly in patients with disorders of the small bowel. In particular the assessment of small obscure gastrointestinal bleeding sources and the detection of shallow inflammatory lesions in the small bowel, have been greatly facilitated by this novel imaging procedure. We report two cases of patients with obscure gastrointestinal bleeding, in whom normal passage of the capsule through the antroduodenal junction was inhibited. This was because of delayed gastric emptying in both patients, which was presumably caused by functional impairment of pyloric motility. To facilitate capsule transport into the small bowel, after swallowing the capsule each patient underwent unsedated upper gastrointestinal endoscopy during which the capsule was grasped with a polypectomy snare, directly transported through the pylorus, and finally released upon arrival in the second portion of the duodenum. Capsule recordings revealed the source of bleeding in both patients and their medical or surgical treatment was subsequently escalated. Capsule imaging of the small bowel facilitated by esophagogastroduodenoscopy (EGD) is safe, and can be applied when patients have functional disorders of pyloric motility. PMID- 12584643 TI - Recurrent nonvariceal upper gastrointestinal bleeding in a patient with gastroduodenal schistosomiasis. AB - In this case report, we describe the rare situation of a patient with nonvariceal upper gastrointestinal bleeding induced by gastric and duodenal involvement of Schistosoma mansoni infection. In this unique case severe, recurrent upper gastrointestinal bleeding was induced by central ulcerations of gastric pseudopolypoid and duodenal polypoid lesions. However, very atypically, there were no signs of portal hypertension, coagulopathy, or variceal bleeding, and no macroscopic evidence of lower gastrointestinal tract involvement. Neither anti ulcer therapy nor endoscopic hemostasis methods were effective in preventing recurrent bleeding episodes. Finally, typical histological and serological tests (positive for S. mansoni hemagglutination) led to the correct diagnosis, and the patient was completely cured by specific antischistosomal therapy. PMID- 12584644 TI - Colorectal stenting. AB - Intestinal stenting is a procedure that is rapidly coming into more widespread use. It is reported to be a safe and highly effective alternative in patients with unresectable intestinal obstruction. Recent reports show the value of this procedure in colorectal obstruction, as it is relatively safe and can avoid the need for high-risk emergency laparotomy and resection in two sessions. New stents can be inserted in a one-step procedure, with delivery through the scope during the initial diagnostic endoscopy. If examinations show that curative treatment is not possible, resection can be completely avoided, so that the initial stent already forms part of the palliative strategy. Although stents are expensive, the procedure appears to be cost-effective, since emergency surgery can be avoided in patients with acute bowel obstruction, and in those with advanced disease no resection of the colon is necessary. PMID- 12584645 TI - Endoscopic ultrasound-guided cystogastrostomy. PMID- 12584646 TI - Conservative vs. endoscopic closure of colocutaneous fistulas after percutaneous endoscopic gastrostomy complications. PMID- 12584648 TI - Endoscopic pancreatic sphincterotomy: dual sphincterotomy vs. endoscopic pancreatic sphincterotomy alone. PMID- 12584650 TI - Gastrobronchial fistula after caustic injury due to lye ingestion. PMID- 12584651 TI - Successful management of postoperative pancreatic fistula by application of constructed S-type pancreatic stent after operation for abnormal biliary pancreatic junction. PMID- 12584652 TI - Sudden asphyxia due to a laryngeal lipoma following esophageal endosonography. PMID- 12584653 TI - Costochondral deformity masquerading as a submucosal gastric tumor. PMID- 12584654 TI - Endoscopic appearance of colonic tuberculosis. PMID- 12584655 TI - [Old age as risk factor for complications of malaria in non-immune travellers]. AB - BACKGROUND: Malaria plays an important role as a dangerous imported infection in Europe. In addition, the number of elderly travellers is increasing. We examined the hypothesis, whether age can be considered as a risk factor for the development of complications in malaria. PATIENTS AND METHODS: We examined retrospectively 134 case records of patients of all age groups with a plasmodial infection. We analysed personal data, travel details and patient histories, the neurological status, respiratory function, as well as haematological, biochemical and parasitological parameters. RESULTS: There is significant evidence for a correlation between age and the frequency of complications in malaria like cerebral involvement, respiratory failure, renal failure, anaemia, and hyperparasitaemia. Out of 132 patients over 15 years of age, 49 (37.1 %) developed a severe course of malaria. In the age group of 60 years and above this percentage increased to 61.5 %. The age distribution in the group with severe malaria was significantly shifted towards the elderly (p = 0.016 in Mann-Whitney test). The duration of hospitalisation also increased with age from an average of five days for the group below 45 years to 21 days for the elderly of 60 years and above. CONCLUSION: Our findings suggest a higher risk for a severe course of malaria in the elderly. For this reason, extensive advice on the correct use of exposure- and chemoprophylaxis of malaria is especially important for travellers above the age of 60. PMID- 12584656 TI - [Results of 20 years of voluntary iodide salt prevention in South Tyrol]. PMID- 12584657 TI - [Arrhythmogenic right ventricular dysplasia as a cause of "sudden cardiac death" with survival]. AB - HISTORY: A 42 year old woman was resuscitated from ventricular fibrillation. 5 months previously she had a syncope. Her nephew had died of sudden cardiac death at the age of 25 years. INVESTIGATIONS: There was no evidence for ST segment elevation, myocardial infarction or pulmonary embolism. The ECG showed right precordial T wave inversion. Coronary artery disease was excluded angiographically. Echocardiography and angiography revealed inferior wall akinesia of the right ventricle with normal left ventricular function and chamber size. Ventricular fibrillation could not be reproduced by programmed stimulation of the right ventricle during an electrophysiologic study. Results of endomyocardial biopsy of the right ventricle showed a focal fibrous infiltration of the myocardium. Magnetic resonance imaging confirmed inferior wall abnormalities of the right ventricle without typical fatty infiltration in the right ventricular myocardium. CLINICAL COURSE: The patient recovered rapidly without neurologic deficits. Arrhythmogenic right ventricular dysplasia was suspected, and a cardioverter defibrillator (ICD) was implanted. Within 6 months after implantation the ICD memory showed no evidence of ventricular fibrillation. CONCLUSION: Arrhythmogenic right ventricular dysplasia is an important cause of ventricular fibrillation with a potential risk of sudden cardiac death in young persons. Concealed arrhythmogenesis as an early manifestation of right ventricular dysplasia is difficult to detect. PMID- 12584658 TI - [Viral anti-interferon strategies: mechanisms and clinical impact]. PMID- 12584659 TI - [Current developments in radiation protection of occupationally exposed persons]. PMID- 12584660 TI - [Risk factor grading for prevention of coronary artery disease]. PMID- 12584661 TI - [Opioids and driving ability -- a problem unsolved?]. PMID- 12584662 TI - [Differential diagnosis of isolated increased SGOT level]. PMID- 12584663 TI - Cellular targets for angiotensin II fragments: pharmacological and molecular evidence. AB - Although angiotensin II has long been considered to represent the end product of the renin-angiotensin system (RAS), there is accumulating evidence that it encompasses additional effector peptides with diverse functions. In this respect, angiotensin IV (Ang IV) formed by deletion of the two N terminal amino acids, has sparked great interest because of its wide range of physiological effects. Among those, its facilitatory role in memory acquisition and retrieval is of special therapeutic relevance. High affinity binding sites for this peptide have been denoted as AT(4)- receptors and, very recently, they have been proposed to correspond to the membrane-associated OTase/ IRAP aminopeptidase. This offers new opportunities for examining physiological roles of Ang IV in the fields of cognition, cardiovascular and renal metabolism and pathophysiological conditions like diabetes and hypertension. Still new recognition sites may be unveiled for this and other angiotensin fragments. Recognition sites for Ang-(1-7) (deletion of the C terminal amino acid) are still elusive and some of the actions of angiotensin III (deletion of the N terminal amino acid) in the CNS are hard to explain on the basis of their interaction with AT(1)-receptors only. A more thorough cross-talk between in vitro investigations on native and transfected cell lines and in vivo investigations on healthy, diseased and transgenic animals may prove to be essential to further unravel the molecular basis of the physiological actions of these small endogenous angiotensin fragments. PMID- 12584664 TI - Between-patient differences in the renal response to renin-angiotensin system intervention: clue to optimising renoprotective therapy? AB - Renin-angiotensin-aldosterone system (RAAS) blockade with angiotensin-converting enzyme inhibitors (ACE-I) or angiotensin II (Ang II), AT(1)-receptor blockers (ARB) is the cornerstone of renoprotective therapy. Still, the number of patients with end-stage renal disease is increasing worldwide, prompting the search for improved renoprotective strategies. In spite of proven efficacy at group level, the long-term renoprotective effect of RAAS blockade displays a marked between patient heterogeneity, which is closely linked to between-patient differences in the intermediate parameters of blood pressure, proteinuria and renal haemodynamics. Of note, the between-patient differences by far exceed the between regimen differences, and thus may provide a novel target for exploration and intervention. The responsiveness to RAAS blockade appears to be an individual characteristic as demonstrated by studies applying a rotation-schedule design. The type and severity of renal disease, obesity, insulin-resistance, glycaemic control, and genetic factors may all be involved in individual differences in responsiveness, as well as dietary factors, such as dietary sodium and protein intake. Several strategies, such as dietary sodium restriction and diuretic therapy, dose-titration for proteinuria, and dual RAAS blockade with ACE-I and ARB, can improve the response to therapy at a group level. However, when analysed for their effect in individuals, it appears that these measures do not allow poor responders to catch up with the good responders, i.e. in spite of their efficacy at group level, the available measures are usually not sufficient to overcome individual resistance to RAAS blockade. We conclude that between-patient differences in responsiveness to renoprotective intervention should get specific attention as a target for intervention. Unravelling of the underlying mechanisms may allow development of specific intervention. Based on the currently available data, we propose that response-based treatment schedules, with a multidrug approach titrated and adapted at individual responses rather than fixed treatment schedules, may provide a fruitful strategy for more effective renoprotection. PMID- 12584665 TI - Local renin-angiotensin systems and their interactions with extra-adrenal corticosteroid production. AB - Adrenal aldosterone production is regulated by the renin-angiotensin system (RAS). It is now known that several other tissues are capable of extra-adrenal aldosterone biosynthesis and that these tissues can also generate angiotensin II through local RAS. Therefore, the regulation of local aldosterone production by the local RAS is a distinct possibility. In this review, we present evidence for the existence of such systems in the vascular system, heart and brain. We then discuss the possibility of interactions between the RAS and aldosterone synthesis at the local level and speculate on the possible physiological effects of such systems in these tissues. PMID- 12584666 TI - Blood pressure and cardiac autonomic function in relation to risk factors and treatment perspectives in Type 1 diabetes. AB - The cumulative incidence of diabetic nephropathy in Type 1 diabetes mellitus is in the order of 25 30%. The recognition that elevated blood pressure (BP) is a major factor in the progression of these patients to end-stage renal failure has led to the widespread use of antihypertensive therapy in order to preserve glomerular filtration rate and ultimately to reduce mortality. The routine measurement of microalbuminuria allows early identification of the subgroup of patients at increased risk of developing clinical nephropathy. Microalbuminuric Type 1 diabetic patients show a number of characteristic pathological abnormalities. In addition to elevated BP and abnormal circadian rhythm, there are also associated abnormalities of vagal function, lipid profile and endothelial function, as well as an increased prevalence of retinopathy. The first section of this two-part review focusses on the early changes associated with renal involvement in Type 1 diabetes. It addresses the associations between urinary albumin excretion, glycaemic control, smoking, BP, circadian BP variation, QT interval abnormalities and autonomic function in three groups of patients; those with normoalbuminuria, those progressing towards microalbuminuria and those with established low-grade microalbuminuria. PMID- 12584667 TI - The renin-angiotensin-aldosterone system and the eye in diabetes. AB - Diabetic retinopathy is the leading cause of blindness in the under 65s, and with the burden of disease case load expected to exceed 200 million worldwide within 10 years, much effort is being spent on prophylactic interventions. Early work focused on improving glycaemic control; however, with the publication of EURODIAB Controlled trial of Lisinopril in Insulin-dependent Diabetes (EUCLID) and United Kingdom Prospective Diabetes Study (UKPDS), the focus has recently moved to control of blood pressure and specifically the renin-angiotensin system (RAS). There is a large body of evidence for a local RAS within the eye that is activated in diabetes. This appears to be directly responsible, as well as indirectly through other mediators, for an increase in concentration of vascular endothelial growth factor (VEGF), a selective angiogenic and vasopermeability factor that is implicated in the pathogenesis of diabetic retinopathy. Inhibition of angiotensin-converting enzyme appears to reduce concentrations of VEGF, with a concurrent anti-proliferative effect independent of systemic VEGF levels or blood pressure. Angiotensin II (Ang II) Type 1 (AT(1)) receptor blockade has been shown to reduce neovascularisation independent of VEGF levels in animal models. This may be due to antagonism of activation of mitogen-activated protein kinase, which is a potent cellular proliferation stimulator, by Ang II, although this needs further evaluation. PMID- 12584668 TI - The pharmacokinetics and pharmacodynamics of angiotensin-receptor blockers in end stage renal disease. AB - Angiotensin-converting enzyme (ACE) inhibitors and more recently angiotensin receptor blockers (ARBs) have become popular therapies in the end-stage renal disease (ESRD) patient. The ability of either of these drug classes to reduce blood pressure in the ESRD patient is well accepted; however, there is considerably less information available to guide the clinician in the safe and effective use of these drugs in the ESRD patient with congestive heart failure and/or coronary artery disease. Head-to-head studies in the ESRD patient are lacking for both drug classes. Several pharmacokinetic factors can influence the selection of these drugs, including dialysability and the propensity for systemic accumulation. ACE inhibitors (ACE-Is) and ARBs are recognised as having a range of nonpressor effects that are pertinent to patients with ESRD. Such effects include their ability to decrease both thirst drive and erythropoiesis. These drug classes, though, are distinguishable by the unique adverse effect profile for ACE-Is. As is the case in patients without renal failure, ESRD patients can experience cough and, less frequently, angioneurotic oedema with ACE-Is. In the ESRD population, so-called anaphylactoid dialyser reactions can occur in conjunction with ACE-I use. The use of a drug from within the ARB class carries both less risk and permits a compound with a preferred pharmacokinetic profile limited dialysability and minimal systemic accumulation to be administered. These attributes would favour the increased use of ARBs in this population. PMID- 12584669 TI - The DIabetic Retinopathy Candesartan Trials (DIRECT) Programme, rationale and study design. AB - The DIabetic Retinopathy Candesartan Trials (DIRECT) Programme consists of three randomised, double-masked, parallel, placebo-controlled studies to determine the impact of treatment with candesartan on diabetic retinopathy. In Type 1 diabetes, 1,700 patients without retinopathy will be randomised into a primary prevention study, and 1,200 with non-proliferative retinopathy into a secondary prevention study. In Type 2 diabetes, 1,600 patients with non-proliferative retinopathy will be randomised. Patients will be followed for at least three years. Eligible patients must be normotensive (systolic blood pressure [SBP] 130 mmHg and diastolic blood pressure [DBP] 85 mmHg) without antihypertensive medication in Type 1 diabetes, and either normotensive or treated hypertensive (SBP 160 mmHg and SBP 90 mmHg) and not taking angiotensin-converting enzyme inhibitors or AT(1) receptor blockers in Type 2 diabetes. All patients will be normoalbuminuric, based on two overnight urine collections. The primary endpoint is based upon retinal photographs, graded to the Early Treatment of Diabetic Retinopathy Study scale. A two-step increase on this scale defines incidence, and a three-step increase defines progression of retinopathy. The main secondary endpoint for each study is change in urinary albumin excretion rate. A positive outcome of the DIRECT Programme would be an important step forward in the clinical management of patients with diabetes. PMID- 12584670 TI - ACE-I and ARBs in early diabetic nephropathy. AB - INTRODUCTION: Antihypertensive treatment of patients with clinical manifestations of diabetic nephropathy, and especially, renin-angiotensin system (RAS) inhibition, slows, but may not fully arrest progression towards end-stage renal disease. Studies using hard endpoints such as doubling of serum creatinine, dialysis, or death that are initiated before emergence of any renal functional abnormalities in diabetes, would be of impractical length and size. We therefore undertook a primary prevention study (The Renin-Angiotensin System Study or RASS) to determine if inhibition of the RAS could slow the development of a key diabetic glomerulopathy structural endpoint, increase in mesangial fractional volume (Vv[Mes/glom]). METHODS: This is a parallel group, double-blind, placebo controlled trial with 285 patients with Type 1 diabetes mellitus (95 per group) randomised to receive the angiotensin-converting enzyme inhibitor, enalapril, the angiotensin II receptor blocker, losartan, or placebo. All patients are normotensive, normoalbuminuric and have normal or increased glomerular filtration rates at study entry. The study is based on primary endpoint of change in Vv(Mes/glom) from baseline to the five-year renal biopsy, with baseline and interval measures of albumin excretion rate, glomerular filtration rate, blood pressure, and glycaemia. Baseline, mid-point, and five-year retinal fundus photography are also performed. RESULTS: One thousand and sixty-five patients were interviewed, 707 refused participation and 73 were excluded. The target of 285 subjects were randomised and their clinical and demographic characteristics are described. Biopsy complications occurred in 17 (6%), only one of which required hospitalisation. There were no permanent biopsy-related sequelae. CONCLUSIONS: Renal structural variables are reasonable surrogate endpoints for studies of progression of early diabetic nephropathy. Although requiring substantial recruitment effort, diabetic nephropathy primary prevention trials based on change in renal structure are feasible. PMID- 12584672 TI - Bovine viral diarrhea virus type 1- and type 2-specific bovine T lymphocyte subset responses following modified-live virus vaccination. AB - Expression of CD25 (interleukin-2 receptor alpha chain) was used to monitor antigen-specific activation of T lymphocyte subsets (CD4+, CD8+, and gamma delta T cells) from cattle immunized with modified-live virus (MLV) bovine viral diarrhea virus (BVDV) vaccines. Two groups of 15 animals each were vaccinated with one dose of either BVDV genotype 1 (BVDV-1) or BVDV-1 and BVDV genotype 2 (BVDV-1/2). Six animals negative for both BVDV antibody and BVDV virus were used as negative controls. Three animals vaccinated 7 and 5 weeks before the start of the experiment with MLV BVDV-1 vaccine served as positive controls. Blood samples were taken from the negative control group, the positive control group, and the BVDV-1/2 group 0, 21, 35, 60, and 90 days after vaccination. Blood samples were taken from the BVDV-1 group 0, 21, and 90 days after vaccination. Isolated peripheral blood lymphocytes from immunized and control animals were incubated for 5 days with and without BVDV-1 or BVDV-2. Compared with nonvaccinated animals, a significant (P <.05) increase in expression of CD25 by CD4+ (60 days), CD8+, and gammadelta T (35 to 90 days) lymphocytes from the group given BVDV-1/2 was detected following in vitro exposure to BVDV-1 or BVDV-2 after vaccination. The CD8+ and gammadelta T cells from the group vaccinated with BVDV-1 had significantly (P <.05) increased expression of CD25 compared with nonvaccinates following postvaccination exposure to in vitro BVDV-1 but not to BVDV-2. There was no significant difference between the two vaccinated groups in CD25 expression on any of the T cell subsets in response to BVDV-1 or BVDV-2 exposure. A single administration of MLV BVDV vaccine may be more effective at stimulating CD8+ and gammadelta T cell-specific immune responses to the homologous genotype than to the heterologous genotype. PMID- 12584671 TI - Effects of the vasopeptidase inhibitor, omapatrilat, in 723 patients with coronary heart disease. AB - INTRODUCTION: Among individuals with a history of myocardial infarction (MI), higher levels of blood pressure (BP) are associated with increased long-term risks of death from coronary heart disease. Treatment with a BP-lowering regimen, based on omapatrilat may result in greater clinical benefits than treatment with a regimen based on a regular angiotensin-converting enzyme (ACE) inhibitor because of more favourable effects on the renin-angiotensin-aldosterone system. METHODS: Seven hundred and twenty-three clinically stable patients with a history of MI or unstable angina, and a mean entry BP of 134/77 mmHg, were randomised to six months treatment with omapatrilat 40 mg, omapatrilat 20 mg, or matching placebo. RESULTS: After six months, mean BP levels (systolic/diastolic) in the omapatrilat 40 mg group were reduced by 4.3/2.9 mmHg (95% confidence interval 1.3 to 7.2/1.2 to 4.6). Mean BP levels in the omapatrilat 20 mg group were reduced by 4.6/1.0 mmHg (1.6 to 7.6/ 0.7 to 2.6) in comparison with the placebo group. Both doses of omapatrilat also produced significant decreases in plasma ACE activity and significant increases in levels of plasma renin activity, atrial natriuretic peptide, endothelin and homocysteine (p<0.05 for all). Premature discontinuations were more frequent with omapatrilat than with placebo (p<0.001 for 20 mg and 40 mg). CONCLUSIONS: Omapatrilat produced changes in BP, neurohormone and biochemical parameters that were similar for the two doses. The long-term clinical implications of the observed effects are uncertain and a large-scale randomised trial would be required to reliably establish the effects of omapatrilat on the risks of major vascular disease events among patients with coronary heart disease. PMID- 12584673 TI - Efficacy of extended pirlimycin hydrochloride therapy for treatment of environmental Streptococcus spp and Staphylococcus aureus intramammary infections in lactating dairy cows. AB - Fifty-one chronically infected lactating dairy cows were used to evaluate the efficacy of extended pirlimycin therapy regimens for treatment of intramammary infections by environmental Streptococcus spp and Staphylococcus aureus. Cows (n = 47) with one or more infected mammary quarters were blocked by parity and randomly allocated to one of three groups for treatment with pirlimycin (50 mg/mammary quarter) as follows: one treatment per day for 2 days (n = 36 infected mammary quarters); one treatment per day for 5 days (n = 36 infected mammary quarters); and one treatment per day for 8 days (n = 20 infected mammary quarters). Four cows with nine infected mammary quarters were included as untreated controls. Milk samples from each mammary quarter were collected 7 days before treatment, immediately before treatment, and weekly for 4 weeks after the final treatment for microbiological evaluation. A bacteriologic cure was defined as a treated, infected quarter that was bacteriologically negative for the presence of previously identified bacteria at weekly intervals after treatment. Efficacy of pirlimycin therapy against intramammary infections caused by environmental Streptococcus spp and S. aureus was 44.4%, 61.1%, and 95.0% for the 2-, 5-, and 8-day treatment regimens, respectively. None of the infections in the untreated control quarters was cured. Significant differences in efficacy were detected between all pirlimycin groups and the untreated control group, between the 8- and 2-day treatment regimens, and between the 8-day and 5-day treatment regimens (P < or = .05). Results of this study indicate that extended pirlimycin therapy was effective in eliminating intramammary infections caused by environmental streptococci and S. aureus in lactating dairy cows. PMID- 12584674 TI - Bacterial cure and somatic cell count response of dairy cows with a positive California Mastitis Test at calving to therapy with cephapirin sodium. AB - Cows without signs of clinical mastitis were evaluated by the California Mastitis Test at calving (Day 0). Milk samples from 117 of 184 quarters (64 cows) were positive by this test for mastitis and were submitted for bacterial culture and determination of somatic cell counts. Cows with infected quarters were randomly allocated to treatment with cephapirin sodium by intramammary infusion or to be untreated as controls. Two and 4 weeks following calving, milk was again sampled from the infected quarters and tested. By the 4-week evaluation, the quarters treated with cephapirin sodium had significantly (P < or = .05) fewer positive bacterial cultures and somatic cell counts were significantly (P < or =.05) reduced compared with untreated control quarters. PMID- 12584675 TI - Efficacy of a 1% lambdacyhalothrin cattle Pour-on (Saber) against sucking and biting lice infesting beef cattle. AB - Four studies were conducted to determine the efficacy of a 1% lambdacyhalothrin pour-on (Saber Pour-on, Schering-Plough Animal Health) for control of sucking lice (Linognathus vituli, Haematopinus eurysternus, Solenopotes capillatus) and biting lice (Damalinia bovis) on beef cattle. Seventy-four mixed-breed cattle naturally infested with one or more species of lice at locations in Wisconsin, Nebraska, Oklahoma, and North Dakota were included in the study. Pretreatment lice samples were taken and identified by genus with the exception that sucking lice were not identified by genus at the North Dakota site. In January or February, half of the cattle at each location received a single application of 1% lambdacyhalothrin pour-on at 10 ml per head for cattle weighing less than 273 kg (600 lb) or 15 ml per head for cattle weighing 273 kg or more. The other cattle at each site served as untreated controls. Lice on designated body areas were counted 2, 4, 6, and 8 weeks after treatment, and the sum of all lice observed on each counting area was reported for each animal. The collective efficacy 6 weeks after treatment against three species of sucking lice (L. vituli, H. eurysternus, S. capillatus) was 88.4% at the Wisconsin site, 92.0% at the North Dakota site, and 100% at the Nebraska and Oklahoma sites. The 1% lambdacyhalothrin pour-on eliminated all biting lice within 2 weeks after treatment, and no biting lice were detected 8 weeks after treatment. A single treatment of 1% lambdacyhalothrin pour-on administered when lice populations were highest (January or February) provided effective season-long control of both biting and sucking lice on cattle. PMID- 12584676 TI - Survey of Neospora caninum antibodies in dairy and beef cattle from five regions of the United States. AB - Serum samples from 4,907 cows from 93 dairy and five beef herds were analyzed for antibodies to Neospora caninum by commercially available ELISA. The herds were recruited from 20 states and the territory of Puerto Rico. A questionnaire assessed the herd's abortion history and exposure risks to N. caninum. Results showed that 788 (16%) cows tested positive for antibodies to N. caninum. At least one positive animal was detected in 90% of the herds tested. The prevalence of cows seropositive to N. caninum varied from 2% to 65% among herds. Abortion rates ranging from 2% to 19% were reported. The epidemiologic data collected from the questionnaire suggested that the presence of farm dogs does not increase the risk of exposure of cows to N. caninum. PMID- 12584677 TI - Determination of plasma and skin concentrations of orbifloxacin in dogs with clinically normal skin and dogs with pyoderma. AB - Plasma and skin concentrations of orbifloxacin (Orbax tablets, Schering-Plough Animal Health) were assessed in 14 clinically normal dogs and 14 dogs with pyoderma following oral administration of the drug at 7.5 mg/kg once daily for 5 to 7 days. Skin biopsies and whole blood samples were obtained before dosing and at the time of the expected maximum concentration in skin (3 hours after dosing) on the first and on the fifth to seventh day of dosing. Skin biopsies and plasma were analyzed for orbifloxacin concentrations by high-performance liquid chromatography. Dogs with pyoderma had significantly higher mean skin concentrations of orbifloxacin within 3 hours of administration (Day 0: 7.80 +/- 3.40 mcg/g, Days 4 to 6: 9.47 +/- 6.23 mcg/g) than did dogs with normal skin (Day 0: 3.85 +/- 1.08 mcg/g, Days 4 to 6: 5.43 +/- 1.02 mcg/g). After dosing on Day 0 and after five to seven daily treatments, dogs with pyoderma had significantly higher mean orbifloxacin skin:plasma ratios (1.40 and 1.44, respectively) than did clinically normal dogs (0.81 and 0.96, respectively). The accumulation of orbifloxacin in diseased skin may contribute to the efficacy of this compound for the treatment of bacterial skin infections. PMID- 12584678 TI - Plasma concentrations of enrofloxacin and its active metabolite ciprofloxacin in dogs following single oral administration of enrofloxacin at 7.5, 10, or 20 mg/kg. AB - Plasma concentrations of enrofloxacin and its active metabolite ciprofloxacin were monitored following oral administration of enrofloxacin at 7.5, 10, and 20 mg/kg to six healthy female bloodhounds using a randomized crossover design. Plasma samples were collected at various times over 24 hours following drug administration. Both the parent drug and its metabolite were detected by high performance liquid chromatography, and plasma drug concentration-versus-time curves were subjected to noncompartmental pharmacokinetic analysis. Descriptive statistics were determined for each dosage, and comparisons were made among dosage groups for selected pharmacokinetic parameters. Increasing dosages of enrofloxacin resulted in increased plasma concentrations of both enrofloxacin and ciprofloxacin. Maximum concentration (Cmax) was 2.12 +/- 0.59, 2.1 +/- 0.34, and 4.74 +/- 1.05 mcg/ml for enrofloxacin and 1.30 +/- 0.31, 1.30 +/- 0.32, and 1.86 +/- 0.35 mcg/ml for ciprofloxacin when enrofloxacin was given at dosages of 7.5, 10, and 20 mg/kg, respectively. Cmax and area under the curve (AUC) for both enrofloxacin and ciprofloxacin were significantly greater at 20 mg/kg than at 7.5 and 10 mg/kg. Disappearance half-life was similar for all dosages, ranging from 4.6 to 5.2 hours for enrofloxacin and 8.8 to 10.7 hours for ciprofloxacin. Ciprofloxacin contributed up to 42% of the Cmax and up to 55% of the AUC of the total (enrofloxacin plus ciprofloxacin). For organisms with a minimum inhibitory concentration (MIC) of 0.5 mcg enrofloxacin/ml, an inhibitory quotient (IQ; Cmax:MIC) of 8 or more was achieved in plasma only at 20 mg/kg. PMID- 12584679 TI - Use of a C6 ELISA test to evaluate the efficacy of a whole-cell bacterin for the prevention of naturally transmitted canine Borrelia burgdorferi infection. AB - A commercially available C6 ELISA kit was used to detect antibodies induced by natural infection with Borrelia burgdorferi in dogs that lived in an area endemic for Lyme disease. Rates of infection were determined both for nonvaccinated dogs and those that had been vaccinated with a whole-cell B. burgdorferi bacterin (Lyme Vax, Fort Dodge Animal Health) before 6 months of age and were boostered annually. Vaccinated dogs had an infection rate of 5% (8 of 163), whereas 64% (25 of 39) of the non-vaccinated dogs were positive for B. burgdorferi antibodies. The preventable fraction, determined by comparing infection rates in unvaccinated and vaccinated dogs, was 92.2% (95% confidence interval: 84.3% to 96.3%). In addition, screening of nonvaccinated dogs at six Connecticut clinics (Middletown, Portland, Essex, Old Lyme, Durham, and Marlborough) with the C6 ELISA test revealed infection rates ranging from 41% to 73%, demonstrating a high level of infected dogs in the area. It was concluded that emphasis should be placed on vaccinating young dogs at risk for Lyme disease before they are exposed to infected ticks. Results of this study support the value of immunization with this whole-cell Lyme disease bacterin for dogs at risk for infection by B. burgdorferi. PMID- 12584680 TI - Effect of perioperative oral carprofen on postoperative pain in dogs undergoing surgery for stabilization of ruptured cranial cruciate ligaments. AB - A randomized, placebo-controlled, parallel study was conducted to investigate the effectiveness of oral carprofen for the control of postoperative pain in dogs undergoing knee surgery for stabilization of ruptured cranial cruciate ligaments. Dogs were randomly assigned to treatment with carprofen (n = 10) or placebo (n = 9). Pain was assessed at 1, 2, 4, 6, 24, and 48 hours and 10 and 21 days postoperatively. Eight of 10 dogs treated with carprofen and five of nine dogs treated with placebo were given at least one dose of morphine as rescue therapy. The mean relative dose of morphine given at 1 hour (P =.01) and 24 hours (P =.02) after surgery was greater for dogs treated with carprofen than for dogs given a placebo. There were no significant postoperative differences in cortisol levels or any measured variable. It appears that the scoring system used was not sensitive enough to detect differences in pain between a known analgesic and a placebo. PMID- 12584681 TI - Efficacy of three dose volumes of topically applied 65% permethrin against Ctenocephalides felis and Rhipicephalus sanguineus on dogs weighing 30 kg or more. AB - Twenty-five dogs were evenly and randomly allocated by weight to five treatment groups: untreated control, 2 ml 65% permethrin (dogs weighing 15 to 29 kg, "average dogs"), and 2, 3, or 4 ml 65% permethrin (dogs weighing > or = 30 kg, "big dogs"). Each dog was infested with 125 unfed, adult cat fleas, Ctenocephalides felis, and 50 unfed, adult brown dog ticks, Rhipicephalus sanguineus, on Days -3 (ticks only), -2 (fleas only), 3, 7, 14, 17, 21, 24, and 28. Fleas and ticks were counted 1 and 3 or 4 days after each infestation. The duration of efficacy (defined as >90%) against C. felis was 28 to 31 days. The efficacy against fleas 31 days after application of 2, 3, or 4 ml on big dogs ranged from 79.1% (2 ml) to 100% (4 ml). Big dogs that received either 3 or 4 ml of 65% permethrin had significantly (P < or = .05) fewer fleas at several evaluations between 15 and 31 days after treatment. The duration of efficacy against R. sanguineus was 15 (2 ml for big dogs) to 28 (2 ml for average dogs) days. The efficacy against R. sanguineus 28 days after treatment ranged from 79.1% (2 ml on big dogs) to 94.1% (2 ml on average dogs). Significantly (P < or = .05) fewer ticks were present at several evaluations after treatment on big dogs that received 3 or 4 ml than were present on big dogs treated with 2 ml. No significant differences were detected between the 3- and 4-ml groups from Days 10 to 31; however, the geometric mean number of ticks in the group treated with 4 ml was numerically lower than that for dogs treated with 3 ml on several occasions. These data indicate that a dose volume of 3 or 4 ml of 65% permethrin is needed to obtain an adequate level and duration of efficacy against both C. felis and R. sanguineus on dogs weighing 30 kg or more. PMID- 12584682 TI - Treatment of periodontal pockets with doxycycline in beagles. AB - Following pretreatment with clindamycin, cleaning, scaling, polishing, and curettage, six beagles that were patients at the Dental Department of the Clinic for Surgery and Ophthalmology of the University of Veterinary Medicine of Vienna received a doxycycline polymer filling (Doxirobe, Pharmacia Animal Health) in periodontal pockets of teeth 204, 208, 304, and 309. Gingivitis index, gingival crevicular fluid, probing depth, and attachment loss were determined before and 6 and 12 weeks after treatment. Teeth 104, 108, 404, and 409 did not receive antibiotic therapy but were pretreated in the same manner as the doxycycline treated teeth. Pocket depth for teeth treated with doxycycline was significantly reduced (improved) by 39% after 6 weeks (P =.001) and by 35% after 12 weeks (P =.001). Pockets around control teeth were improved after cleaning and curettage but were still significantly deeper than around teeth treated with doxycycline. Compared with control teeth, teeth treated with doxycycline had significantly less gingival crevicular fluid after treatment (P =.001). Teeth treated with doxycycline gained significant attachment after 6 (42%) and 12 (38%) weeks. Significantly fewer bacteria were harvested from doxycycline-treated teeth than from control teeth. The gingival index was significantly lower in the doxycycline treated teeth than in the control teeth 6 (P =.002) and 12 (P =.007) weeks after treatment. Local application of doxycycline complements traditional subgingival curettage therapy in a reasonable and effective way and can significantly improve treatment success, especially with regard to pocket depth reduction and attachment gain. PMID- 12584684 TI - Concurrent administration of water-soluble vitamin E can increase the oral bioavailability of cyclosporine a in healthy dogs. AB - Randomized crossover studies were performed to determine the effect of coadministration of d-alpha-tocopheryl polyethylene glycol 1,000 succinate (vitamin E TPGS) on the oral bioavailability of cyclosporine A (CsA) formulated as either Sandimmune (Novartis Pharmaceuticals) or Neoral (Novartis Pharmaceuticals). Healthy dogs were given a single oral dose of each cyclosporine formulation with and without vitamin E TPGS. Blood samples were collected from each dog before and at various intervals for 24 hours after drug administration. Whole-blood CsA concentrations were determined by high-pressure liquid chromatography. Noncompartmental pharmacokinetic analysis showed that coadministration of vitamin E TPGS increased the oral bioavailability of Sandimmune. The bioavailability of Neoral was greater than that of cyclosporine. Concurrent administration of vitamin E TPGS had no consistent effect on the bioavailability of Neoral. PMID- 12584683 TI - Effect of deracoxib, a new COX-2 inhibitor, on the prevention of lameness induced by chemical synovitis in dogs. AB - Twenty-four healthy, mixed-breed hound-type dogs were evenly and randomly assigned to a placebo control group, one of four dosages of deracoxib (0.3, 1, 3, or 10 mg/kg), or carprofen (2.2 mg/kg). Oral dosing of placebo, carprofen, or deracoxib was done 30 minutes before intraarticular injection of urate crystal suspension for induction of synovitis. Ground reaction forces, subjective clinical lameness scores, pain, joint effusion, and quantitative pain threshold responses were measured in a blinded fashion before induction of synovitis and 2, 4, 6, 8, 12, and 24 hours after injection. The medium and high dosages of deracoxib were effective in preventing lameness and pain associated with synovitis. Carprofen was also somewhat effective in attenuating the severity of urate-induced synovitis but to a lesser degree than the medium dose of deracoxib. Preemptive deracoxib treatment at dosages as low as 1 mg/kg reduced lameness and pain of synovitis associated with intraarticular administration of urate crystals. PMID- 12584685 TI - S-adenosylmethionine and cAMP confer differential cytoprotection against bile acid-induced apoptosis in canine renal tubular cells and primary rat hepatocytes. AB - Cholestasis results in the accumulation of cytotoxic bile acids in the body. The cytoprotective effect of S-adenosylmethionine (SAMe) and cAMP were compared in two in vitro models of bile acid-induced apoptosis. Primary cultures of rat hepatocytes and canine renal tubular cells (Madin-Darby canine kidney [MDCK] cells stably transfected with the conjugated bile salt transporter, sodium [Na+]/taurocholate cotransporting polypeptide [Ntcp]) were treated with conjugated bile acids and monitored for apoptosis. Glycine-conjugated bile acids caused similar amounts of apoptosis, whereas taurine-conjugated bile acids were five to 10 times more toxic in MDCK-Ntcp cells than in hepatocytes. Treatment with the 1,4-butanedisulfonate salt of SAMe (500 microM) or the stable chlorophenylthio-cAMP analogue (100 microM) inhibited bile acid-induced apoptosis in hepatocytes by 70% and 40%, respectively. In MDCK-Ntcp cells, SAMe inhibited apoptosis by 20%, but cAMP was without effect. Immunoblotting for activation of putative survival kinases in cAMP-treated cells (phosphorylated protein kinase B [Akt] or mitogen-activated protein kinase [MAPK]) was done using phosphospecific antibodies. cAMP increased Akt phosphorylation threefold in hepatocytes but not in MDCK-Ntcp cells. Activation of p42/p44 MAPK was inhibited by cAMP in both cells. SAMe protects against bile acid-induced apoptosis in hepatocytes and MDCK Ntcp cells. The cytoprotective effect of cAMP is seen only in hepatocytes and may reflect tissue-specific activation of Akt. PMID- 12584686 TI - Evaluation of 65% permethrin spot-on for prevention of canine visceral leishmaniasis: effect on disease prevalence and the vectors (Diptera: Psychodidae) in a hyperendemic area. AB - A study was designed to examine the effect of 65% permethrin spot-on on the prevalence of canine visceral leishmaniasis and the abundance of sand flies in two neighborhoods in Corumba, Mato Grosso do Sul, Brazil known to have a high prevalence of visceral leishmaniasis. An enrollment survey was conducted to determine the prevalence of visceral leishmaniasis. Area residents were provided with information about the project; the study area was defined, and all dogs (160 in Cristo Redentor and 230 in Popular Velha) identified in the study area were enrolled. Three 65% permethrin spot-on treatments (June 15-25, July 13-15, and August 10-12) were administered to 150, 110, and 99 dogs, respectively, in Popular Velha, according to label recommendations. Dogs in Cristo Redentor were untreated controls. Visceral leishmaniasis was diagnosed periodically by indirect immunofluorescence assay. A reduction in canine visceral leishmaniasis prevalence was observed at the Popular Velha site. The infection rate for treated dogs 1 month following the final treatment was approximately 50% reduced from that observed before treatment(19.3% vs 9.6%). Conversely, the infection rate at the control site was more than 80% higher at the September sampling than that observed pretreatment (4.1% vs 7.4%). Similar numbers of sand flies were captured and identified from both sites throughout the study. The results suggest that regular use of 65% permethrin during months of high risk for canine visceral leishmaniasis can be a useful strategy for reducing the prevalence of this disease in hyperendemic areas. It should be stressed, however, that the success of this strategy depends not only on the efficacy of the product itself but also on the adoption of other control measures and on economic variables, considering the low purchasing power of the populations living in higher-risk neighborhoods. PMID- 12584687 TI - Development of resistant bacteria isolated from dogs with otitis externa or urinary tract infections after exposure to enrofloxacin in vitro. AB - Minimum inhibitory concentrations for enrofloxacin were determined for 63 bacterial isolates from dogs with otitis externa or urinary tract infections. Development of resistant mutants was determined after exposing the isolates to enrofloxacin in vitro for up to five serial passages. Results indicated that Pseudomonas aeruginosa and Enterococcus spp isolates exposed to enrofloxacin developed resistance rapidly, whereas Klebsiella, Proteus, and Streptococcus spp were less likely to develop resistance. Despite the presence of enrofloxacin pressure, no resistant bacteria developed in the Escherichia coli and staphylococcal isolates. In many isolates, susceptibility patterns changed from susceptible to intermediate. PMID- 12584689 TI - Preclinical pharmacology of zoledronic acid. AB - The evolution of bisphosphonates over the past 30 years has led to the development of nitrogen-containing bisphosphonates with ever-increasing potency. Recent studies have begun to shed light on the unique mechanism of action and pharmacologic properties of these compounds. On the basis of in vitro studies and animal models of osteoclast-mediated bone resorption, zoledronic acid is the most potent bisphosphonate among a large number of compounds tested, including pamidronate and most other commercially available bisphosphonates. Zoledronic acid maintains bone mass in estrogen-deficient animals without adversely affecting bone mineralization. Moreover, the high potency of zoledronic acid translates into dramatic suppression of bone resorption markers at very low doses in patients with bone metastases, and zoledronic acid has shown efficacy across a broad range of tumor types. Preclinical studies have also shown the potential of bisphosphonates to inhibit tumor cell growth and colonization of the bone and to reduce skeletal tumor burden in animal models. A variety of mechanisms have been proposed to explain these observations and continue to be investigated in animal models. Studies are ongoing to determine if the antitumor potential of bisphosphonates can be exploited in the clinical setting. PMID- 12584690 TI - Treatment of hypercalcemia of malignancy with bisphosphonates. AB - Hypercalcemia of malignancy (HCM) is a potentially life-threatening complication of cancer resulting from increased bone resorption by osteoclasts. Clinical management of HCM primarily consists of intravenous rehydration therapy combined with pharmaceutical agents that decrease osteoclast activity. Several generations of drugs designed to combat HCM have evolved, the latest being intravenous bisphosphonate therapy. Bisphosphonates safely and effectively decrease serum calcium levels by interfering with osteoclast activity and stimulating osteoclast apoptosis. Zoledronic acid is the most recent bisphosphonate approved by the US Food and Drug Administration for treatment of HCM and is significantly more effective in reducing serum calcium levels than previously used bisphosphonates. Furthermore, zoledronic acid has a safety profile similar to that of pamidronate and other intravenous bisphosphonates. Thus, zoledronic acid represents a significant therapeutic advance in the clinical management of HCM. PMID- 12584691 TI - Treatment of bone complications in advanced prostate cancer: rationale for bisphosphonate use and results of a phase III trial with zoledronic acid. AB - Prostate cancer often metastasizes to bone during disease progression. Patients who develop bone metastases have a high risk of developing skeletal complications, including pathologic fractures, spinal cord compression, and severe bone pain. Bisphosphonate therapy is widely used for the prevention of skeletal complications in patients with bone lesions from multiple myeloma and breast cancer. Until recently, however, no bisphosphonate had ever shown objective clinical benefit in patients with prostate cancer and osteoblastic bone lesions. A recent multicenter, randomized, placebo-controlled trial found zoledronic acid (4 mg) to be a safe and effective therapy in patients with bone metastases from hormone-refractory prostate cancer. Zoledronic acid significantly reduced the proportion of patients who experienced skeletal complications and extended the time to first skeletal complication. Further, zoledronic acid significantly reduced the risk of skeletal complications over this 15-month study and provided consistent reductions in bone pain that were significant at the 3- and 9-month time points compared with placebo. These results suggest that zoledronic acid may become an important advancement in the care of patients with prostate cancer metastatic to bone. The role of zoledronic acid in the treatment of patients with prostate cancer continues to evolve. PMID- 12584692 TI - Efficacy and safety of zoledronic acid in the treatment of bone metastases associated with lung cancer and other solid tumors. AB - Advanced solid tumors are often aggressive and recurrent, and overall survival remains relatively poor despite contemporary therapeutic interventions. Bone metastases are common in these patients, and skeletal-related events, including bone pain, pathologic fractures, and potentially life-threatening hypercalcemia of malignancy, undermine the quality of patient survival. Bisphosphonates are widely used in the treatment of bone metastases associated with breast cancer and multiple myeloma, but have not been extensively investigated in the treatment of patients with solid tumors other than breast or prostate cancer. However, a new generation nitrogen-containing bisphosphonate, zoledronic acid, has shown significant clinical benefits in indications in which other bisphosphonates have failed. In a phase III clinical trial in patients with bone metastases from solid tumors other than breast or prostate cancer, treatment with zoledronic acid (4 mg via 15-minute infusion) was well tolerated and significantly decreased the incidence of skeletal-related events and increased the time to first skeletal related event compared with placebo-treated patients. This was the first demonstration of palliative efficacy for bisphosphonate therapy in patients with bone metastases from a wide variety of solid tumors. PMID- 12584693 TI - The antitumor potential of bisphosphonates. AB - Bisphosphonates are primarily known for their ability to inhibit osteoclast mediated bone resorption and to slow the release of calcium and other bone minerals into the blood stream. However, recent preclinical research has shown that bisphosphonates also exhibit potent antitumor activity. Bisphosphonates reduce proliferation and induce apoptosis of tumor cell lines, and they inhibit tumor cell adhesion and invasion of the extracellular matrix in vitro. In addition, bisphosphonates have been shown to reduce skeletal tumor burden in a variety of animal models. This may reflect either direct antitumor effects or indirect effects via osteoclast inhibition and alteration of the bone microenvironment. The complex mechanisms by which bisphosphonates inhibit bone resorption and interfere with tumor growth and metastasis to bone are beginning to be understood. Research is ongoing to fully elucidate these biochemical mechanisms, and well-designed clinical trials are planned to investigate the antitumor potential of modern nitrogen-containing bisphosphonates in the clinical setting. PMID- 12584694 TI - Bisphosphonates for the prevention of bone metastases. AB - Preclinical studies have provided evidence that bisphosphonates have antitumor activity. Bisphosphonates have been shown to induce tumor cell apoptosis in vitro and reduce skeletal tumor burden in tumor xenograft models. Indirect evidence of bisphosphonate antitumor activity has been acquired from studies of clodronate, pamidronate, and zoledronic acid in patients with established bone metastases. Furthermore, clinical studies of daily oral clodronate adjuvant therapy suggest that bisphosphonates can prevent bone metastases, but their effect on overall survival is less certain. Pamidronate and zoledronic acid, potent nitrogen containing bisphosphonates, have shown antitumor activity at approximately 10- to 100-fold lower concentrations than clodronate in vitro and may hold promise for the prevention of bone metastases. However, before the clinical application of bisphosphonates for the prevention of bone metastases can be initiated, clinical trials must assess the optimal stage of disease at which to initiate treatment, the duration of therapy, and the optimal dose and schedule to achieve clinically meaningful antitumor effects. Adjuvant studies of zoledronic acid in patients with breast cancer are in development and should further clarify the role of zoledronic acid in the prevention of bone metastases. PMID- 12584697 TI - Phase separation in metal oxides. AB - A fascinating phenomenon, recently found to occur in certain transition-metal oxides, is phase separation wherein pure, nominally monophasic oxides of transition metals with well-defined compositions separate into two or more phases over a specific temperature range. Such phase separation is entirely reversible, and is generally the result of a competition between charge-localization and delocalization, the two situations being associated with contrasting electronic and magnetic properties. Coexistence of more than one phase, therefore, gives rise to electronic inhomogeneity and a diverse variety of magnetic, transport, and other properties, not normally expected of the nominal monophasic composition. An interesting feature of phase separation is that it covers a wide range of length scales anywhere between 1-200 nm. While cuprates and manganates, especially the latter, provide excellent examples of phase separation, it is possible that many other transition-metal compounds with extended structures will be found to exhibit phase separation. PMID- 12584698 TI - tert-Butylphosphonic acid: from the bulk to the gas phase. AB - The structure of tert-butylphosphonic acid in the solid, in solution, and in the gas phase was studied by single-crystal X-ray diffraction, (1)H and (31)P NMR spectroscopic studies in solution, solid-state (31)P NMR spectroscopy, and electrospray ionization mass spectrometry. In addition, density functional theory (DFT) calculations at the B3LYP/6-31G*, B3LYP/6-31+G*, and B3LYP/6-311+G* level of theory for a large number of H-bonded aggregates of the type (tBuPO(3)H(2))(n) (C(n), P(n); n=1-7) support the experimental work. Crystallization of tBuPO(3)H(2) from polar solvents such as CH(3)CN or THF gives the H-bonded one dimensional polymer 2, whereas crystallization from the less polar solvent CDCl(3) favors the formation of the H-bonded cluster (tBuPO(3)H(2))(6).CDCl(3) (1). In CDCl(3) the hexamer (tBuPO(3)H(2))(6) (C(6)) is replaced by smaller aggregates down to the monomer with decreasing concentration. DFT calculations and natural bond orbital (NBO) analyses for the clusters C(1)-C(7) and the linear arrays P(1)-P(7) reveal the hexamer C(6) to be the energetically favored structure resulting from cooperative strengthening of the hydrogen bonds in the H bonded framework. However, the average hydrogen bond strengths calculated for C(6) and P(2) do not differ significantly (42-43 kJ mol(-1)). The average distances r(O.O), r(Obond;H), r(Pdbond;O), and r(Pbond;OH) in C(1)-C(7) and P(1) P(7) are closely related to the hydrogen bond strength. Electrospray ionization mass spectrometry shows the presence of different anionic species of the type [(tBuPO(3)H(2))(n)-H](-) (A(1)-A(7), n=1-7) depending on the instrumental conditions. DFT calculations at the B3LYP/6-31G* level of theory were carried out for A(1)-A(6). We suggest the dimer [(tBuPO(3)H(2))(2)-H](-) (A(2)) and the trimer [(tBuPO(3)H(2))(3)-H](-) (A(3)) are the energetically favored anionic structures. A hydrogen bond energy of approximately 83 kJ mol(-1) was calculated for A(2). Electrospray ionization mass spectrometry is not suitable to study the assembling process of neutral H-bonded tert-butylphosphonic acid since the removal of a proton from the neutral aggregates has a large influence on the hydrogen bond strength and the cluster structure. PMID- 12584699 TI - Rapid screening of binding constants by calibrated competitive 1H NMR spectroscopy. AB - A calibrated competitive NMR method has been developed that is appropriate for the rapid screening of binding constants. This method involves the initial characterisation of a receptor-substrate binding event for which the (1)H NMR spectrum of a given receptor (calibrant) is modified by the substrate of interest at a range of concentrations. For all subsequent "unknown" receptors, K(a) values are then determined by using a competition assay (in the presence of the calibrant receptor) by measuring a single standard (1)H NMR spectrum. This enables a rapid assessment of the recognition properties of a library of potential receptors. Only the calibrant receptor needs to be NMR active, while the library of putative receptors, as well as the substrate, can be NMR silent. This method assumes the formation of complexes of 1:1 stoichiometry. To demonstrate this methodology, the binding of a number of crown ether type compounds with K+ ions has been studied. Comparison of the binding strengths obtained by using this approach with those in the literature shows excellent agreement. A range of new compounds that have recently been synthesised within our group has also been screened in order to illustrate how this approach can rapidly assess binding ability. This method has significance for chemists working in the fields of combinatorial receptor/substrate design and supramolecular chemistry as a means of rapid optimisation of binding strength. PMID- 12584700 TI - Synthesis and crystal structure of IM-6, a new open framework cobalt-gallium phosphate with ten- and twelve-membered pore openings. AB - A new three-dimensional microporous cobalt-gallium phosphate, named IM-6, has been synthesized under solvothermal conditions with an N-substituted piperazine as organic template. The structure was solved by single-crystal X-ray diffraction (triclinic, P(-)1, a=9.848(20), b=12.470(32), c=12.603(28)A, alpha=63.47(16) degrees, beta=74.56(16) degrees, gamma=76.03(17) degrees). IM-6 exhibits a new framework topology. The inorganic framework is built up of MO(4) (M=Co, Ga) and PO(4) tetrahedra. It displays a two-dimensional interconnected channel system running along the [0(-)11] and [100] directions and delimited by ten- and twelve membered ring openings, respectively. PMID- 12584701 TI - Iron twin-coronet porphyrins as models of myoglobin and hemoglobin: amphibious electrostatic effects of overhanging hydroxyl groups for successful CO/O2 discrimination. AB - Inspired by the observation of polar interactions between CO and O(2) ligands and the peptide residues at the active site of hemoglobin and myoglobin, we synthesized two kinds of superstructured porphyrins: TCP-IM, which contains a linked imidazole ligand, and TCP-PY, which contains a linked pyridine ligand, and examined the thermodynamic, kinetic, and spectroscopic (UV/Vis, IR, NMR, and resonance Raman) properties of their CO and O(2) complexes. On both sides of each porphyrin plane, bulky binaphthyl bridges form hydrophobic cavities that are suitable for the binding of small molecules. In the proximal site, an imidazole or pyridine residue is covalently fixed and coordinates axially to the central iron atom. In the distal site, two naphtholic hydroxyl groups overhang toward the center above the heme. The CO affinities of TCPs are significantly lower than those of other heme models. In contrast, TCPs have moderate O(2) binding ability. Compared with reported model hemes, the binding selectivity of O(2) over CO in TCP-IM and TCP-PY complexes is greatly improved. The high O(2) selectivity of the TCPs is mainly attributable to a low CO affinity. The comparison of k(on)(CO) values of TCPs with those of unhindered hemes indicates the absence of steric hindrance to the intrinsically linear CO coordination to Fe(II) in TCP-IM and TCP PY. The abnormally large k(off)(CO) values are responsible for the low CO affinities. In contrast, k(off)(O(2)) of TCP-PY is smaller than those of other pyridine-coordinated model hemes. For the CO adducts of TCPs, unusually low nu(Fe CO) and unusually high nu(C-O) frequencies are observed. These results can be ascribed to decreased back-bonding from the iron atom to the bound CO. The lone pairs of the oxygen atoms of the hydroxyl groups prevent back-bonding by exertion of a strong negative electrostatic interaction. On the other hand, high nu(Fe O(2)) frequencies are observed for the O(2) adducts of TCPs. In the resonance Raman (RR) spectrum of oxy-TCP-IM, we observed simultaneous enhancement of the Fe O(2) and O-O stretching modes. Furthermore, direct evidence for hydrogen bonding between the hydroxyl groups and bound dioxygen was obtained by RR and IR spectroscopy. These spectroscopic data strongly suggest that O(2) and CO binding to TCPs is controlled mainly by the two different electrostatic effects exerted by the overhanging OH groups: destabilization of CO binding by decreasing back bonding and stabilization of O(2) binding by hydrogen bonding. PMID- 12584702 TI - Determination of the C4-H bond dissociation energies of NADH models and their radical cations in acetonitrile. AB - Heterolytic and homolytic bond dissociation energies of the C4-H bonds in ten NADH models (seven 1,4-dihydronicotinamide derivatives, two Hantzsch 1,4 dihydropyridine derivatives, and 9,10-dihydroacridine) and their radical cations in acetonitrile were evaluated by titration calorimetry and electrochemistry, according to the four thermodynamic cycles constructed from the reactions of the NADH models with N,N,N',N'-tetramethyl-p-phenylenediamine radical cation perchlorate in acetonitrile (note: C9-H bond rather than C4-H bond for 9,10 dihydroacridine; however, unless specified, the C9-H bond will be described as a C4-H bond for convenience). The results show that the energetic scales of the heterolytic and homolytic bond dissociation energies of the C4-H bonds cover ranges of 64.2-81.1 and 67.9-73.7 kcal mol(-1) for the neutral NADH models, respectively, and the energetic scales of the heterolytic and homolytic bond dissociation energies of the (C4-H)(.+) bonds cover ranges of 4.1-9.7 and 31.4 43.5 kcal mol(-1) for the radical cations of the NADH models, respectively. Detailed comparison of the two sets of C4-H bond dissociation energies in 1 benzyl-1,4-dihydronicotinamide (BNAH), Hantzsch 1,4-dihydropyridine (HEH), and 9,10-dihydroacridine (AcrH(2)) (as the three most typical NADH models) shows that for BNAH and AcrH(2), the heterolytic C4-H bond dissociation energies are smaller (by 3.62 kcal mol(-1)) and larger (by 7.4 kcal mol(-1)), respectively, than the corresponding homolytic C4-H bond dissociation energy. However, for HEH, the heterolytic C4-H bond dissociation energy (69.3 kcal mol(-1)) is very close to the corresponding homolytic C4-H bond dissociation energy (69.4 kcal mol(-1)). These results suggests that the hydride is released more easily than the corresponding hydrogen atom from BNAH and vice versa for AcrH(2), and that there are two almost equal possibilities for the hydride and the hydrogen atom transfers from HEH. Examination of the two sets of the (C4-H)(.+) bond dissociation energies shows that the homolytic (C4-H)(.+) bond dissociation energies are much larger than the corresponding heterolytic (C4-H)(.+) bond dissociation energies for the ten NADH models by 23.3-34.4 kcal mol(-1); this suggests that if the hydride transfer from the NADH models is initiated by a one electron transfer, the proton transfer should be more likely to take place than the corresponding hydrogen atom transfer in the second step. In addition, some elusive structural information about the reaction intermediates of the NADH models was obtained by using Hammett-type linear free-energy analysis. PMID- 12584703 TI - Stabilities and isomeric equilibria in aqueous solution of monomeric metal ion complexes of adenosine 5'-diphosphate (ADP3-) in comparison with those of adenosine 5'-monophosphate (AMP2-). AB - Under experimental conditions in which the self-association of the adenine phosphates (AP), that is, of adenosine 5'-monophosphate (AMP(2-)) and adenosine 5'-diphosphate (ADP(3-)), is negligible, potentiometric pH titrations were carried out to determine the stabilities of the M(H;AP) and M(AP) complexes where M(2+)=Mg(2+), Ca(2+), Sr(2+), Ba(2+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), or Cd(2+) (25 degrees C; I=0.1 M, NaNO(3)). It is concluded that in the M(H;AMP)(+) species M(2+) is bound at the adenine moiety and in the M(H;ADP) complexes at the diphosphate unit; however, the proton resides in both types of monoprotonated complexes at the phosphate residue. The stabilities of nearly all the M(AMP) and M(ADP)(-) complexes are significantly larger than what is expected for a sole coordination of M(2+) to the phosphate residue. This increased complex stability is attributed, in agreement with previous (1)H NMR shift studies and further information existing in the literature, to the formation of macrochelates of the phosphate-coordinated metal ions with N7 of the adenine residues. On the basis of recent measurements with simple phosphate monoesters and phosphonate ligands (R MP(2-)) as well as with diphosphate monoesters (R-DP(3-)), where R is a noncoordinating and noninhibiting residue, the increased stabilities of the M(AMP) and M(ADP)(-) complexes due to the M(2+)-N7 interaction could be evaluated and the extent of macrochelate formation calculated. The results show that the formation degrees of the macrochelates for the complexes of the alkaline earth ions are small (about 15 % at the most), whereas for the 3d metal ions as well as for Zn(2+) and Cd(2+) the formation degrees vary between about 15 % (Mn(2+)) and 75 % (Ni(2+)) with values of about 40 and 50 % for Zn(2+) and Cu(2+), respectively. It is interesting to note, taking earlier results for M(ATP)(2-) complexes also into account (ATP(4-)=adenosine 5'-triphosphate), that for a given metal ion in nearly all instances the formation degrees of the macrochelates are within the error limits the same for M(AMP), M(ADP)(-) and M(ATP)(2-) complexes; except for Co(2+) and Ni(2+) it holds M(AMP) > M(ADP)(-) approximately M(ATP)(2 ). This result is astonishing if one considers that the absolute stability constants of these complexes, which are determined largely by the affinity of the phosphate residues, can differ by more than two orders of magnitude. The impact and conclusions of these observations for biological systems are shortly lined out. PMID- 12584705 TI - Synthesis and nonlinear optical absorption of novel porphyrin-osmium-cluster complexes. AB - Reaction of azido(tetra-p-tolylporphyrinato)indium(III) [TTPInN(3)] and [Os(3)(mu H)(2)(CO)(10)] in toluene at 80 degrees C overnight gave two major products, complexes 1 and 2. Complex 1 had an axial bridge of "NH", while 2 had an axial bridge of "N" between the porphyrin and osmium cluster moieties. Complex 1 could be converted to 2 when refluxed in toluene. These two novel porphyrin-osmium clusters are the first axially linked porphyrin-metal cluster complexes. UV/Vis spectroscopy revealed the significant ground state electronic perturbation in the capped complex 2, demonstrating that the remarkable electronic interaction of the moieties within the molecule was achieved by this special structural arrangement. In addition, the electrochemistry of 1 and 2 were investigated and their oxidation current voltage curves are similar to those of indium(III)-porphyrins with a metal-metal sigma bond such as [TPPInRe(CO)(5)] (TPP=tetraphenylporphyrin). The two new molecules also exhibit large nonlinear optical absorption at 532 nm with a ns pulse laser and are potential optical limiting materials for sensor protection in the visible region. PMID- 12584704 TI - Fluorogenic stereochemical probes for transaldolases. AB - Transaldolase catalyzes the transfer of dihydroxyacetone from, for example, fructose 6-phosphate to erythrose 4-phosphate. As a potential probe for assaying fluorescent transaldolase, 6-O-coumarinyl-fructose (1) was prepared in six steps from D-fructose. The corresponding 6-O-coumarinyl-5-deoxy derivative 2 was prepared stereoselectively from acrolein and tert-butyl acetate by a chemoenzymatic route involving Amano PS lipase for the kinetic resolution of tert butyl 3-hydroxypent-4-enoate (7) and E. coli transketolase for assembly of the final product. The corresponding stereoisomer related to D-tagatose was obtained by a chemical synthesis starting from D-ribose. Indeed, transaldolases catalyze the retro-aldolization of substrate 1 to give dihydroxyacetone and 3-O-coumarinyl glyceraldehyde. The latter primary product undergoes a beta-elimination in the presence of bovine serum albumin (BSA) to give the strongly fluorescent product umbelliferone. A similar reaction is obtained with the 5-deoxy analogue 2, but there is almost no reaction with its stereoisomer 3. The stereoselectivity of transaldolases can be readily measured by the relative rates of fluorescence development in the presence of the latter pair of diastereomeric substrates. PMID- 12584706 TI - Generation of reactive species by one-electron reduction of Fischer-type carbene complexes of group 6 metals and their use for carbon-carbon bond formation. AB - Carbon-carbon bond-forming reactions mediated by one-electron reduction of Fischer-type carbene complexes of Group 6 metals were investigated. In the case of aryl- or silylcarbene complexes of tungsten, the anion radical species generated by one-electron reduction smoothly underwent addition reaction to ethyl acrylate. One-electron reduction of alpha,beta-unsaturated carbene complexes afforded biscarbene complexes by dimerization of the corresponding anion radical species at the position gamma to the metal center. In contrast, one-electron reduction of chromium phenyl- or alkylcarbene complexes gave, via carbonyl insertion, alpha-methoxyacylchromate complexes, which further underwent conjugate addition to various electron-poor olefins to give the corresponding alpha methoxyketones. PMID- 12584707 TI - Swelling and shrinking of polyelectrolyte microcapsules in response to changes in temperature and ionic strength. AB - Swelling and shrinking of polyelectrolyte microcapsules consisting of poly(styrene sulfonate, sodium salt) (PSS) and poly(diallyldimethyl ammonium) chloride (PDADMAC) multilayers have been observed in response to temperature and electrolyte exposure, respectively. Heat-induced capsule swelling and capsule wall volume reduction were observed by confocal laser scanning microscopy (CLSM) and scanning force microscopy (SFM). On the other hand, pronounced shrinking in diameter induced by exposure to an electrolyte was observed in parallel to increases in the thickness of the capsule wall. The estimated wall volume was reduced to two thirds of the control for the salt-exposed capsules and one half for the salt-exposed and simultaneously annealed capsules. This reduction in volume was supposedly mainly caused by the compression of the capsule wall due to the ionic screening from the electrolyte. The highly porous microstructure of the multilayers and loosely bound PSS/PDADMAC complex are thought to be responsible for the structure of the PSS/PDADMAC capsules being easily modulated upon annealing and salt-exposure. PMID- 12584708 TI - Hierarchical self-assembly, coassembly, and self-organization of novel liquid crystalline lattices and superlattices from a twin-tapered dendritic benzamide and its four-cylinder-bundle supramolecular polymer. AB - The synthesis and structural analysis of the twin-dendritic benzamide 10, based on the first-generation, self-assembling, tapered dendrons 3,4,5-tris(4' dodecyloxybenzyloxy)benzoic acid and 3,4,5-tris(4'-dodecyloxybenzyloxy)-1 aminobenzene, and the polymethacrylate, 20, which contains 10 as side groups, are presented. Benzamide 10 self-assembles into a supramolecular cylindrical dendrimer that self-organizes into a columnar hexagonal (Phi(h)) liquid crystalline (LC) phase. Polymer 20 self-assembles into an imperfect four-cylinder bundle supramolecular dendrimer, and creates a giant vesicular supercylinder that self-organizes into a columnar nematic (N(c)) LC phase which displays short-range hexagonal order. In mixtures of 20 and 10, 10 acts as a guest and 20 as a host to create a perfect four-cylinder-bundle host-guest supramolecular dendrimer that coorganizes with 10. A diversity of Phi(h), simple rectangular columnar (Phi(r s)) and centered rectangular columnar (Phi(r-c)), superlattices are produced at different ratios between 20 and 10. This diversity of LC lattices and superlattices is facilitated by the architecture of the twin-dendritic building block, polymethacrylate, the host-guest supramolecular assembly, and by hydrogen bonding along the center of the supramolecular cylinders generated from 10 and 20. PMID- 12584709 TI - Synthesis and characterization of luminescent rod-coil block copolymers by atom transfer radical polymerization: utilization of novel end-functionalized terfluorenes as macroinitiators. AB - Two novel, rigid, photoluminescent, substituted terfluorene derivatives were synthesized by utilizing direct bromination and Suzuki coupling reactions. These oligomers were used as initiators for the atom transfer radical polymerization (ATRP) of styrene and tert-butyl acrylate. Thus, diblock and triblock rod-coil block copolymers were prepared with well-defined structure, as far as their size and shape is concerned. Molecular weights up to approximately 21 000 and polydispersity indices not exceeding 1.5 in most cases were obtained. The copolymers emit blue light in solution, and their luminescence properties remain practically invariable when passing from solution to the solid state. No ground state aggregation or excimer formation were observed in the solid state, even after annealing at high temperatures. PMID- 12584710 TI - Fragmentation of transition metal carbonyl cluster anions: structural insights from mass spectrometry. AB - The anionic clusters [HOs(5)(CO)(15)](-), [PtRu(5)C(CO)(15)](2-), [Os(10)C(CO)(24)](2-), [Os(17)(CO)(36)](2-), [Os(20)(CO)(40)](3-), [Co(6)C(CO)(15)](2-), [Pt(3)Ru(10)C(2)(CO)(32)](2-) and [Pd(6)Ru(6)(CO)(24)](2-) have been analysed by energy-dependent electrospray ionisation mass spectrometry (EDESI-MS). Three main features have emerged. Firstly, carbonyl ligands are fragmented from clusters with compact metal cores in an orderly fashion, with each of the ions generated by CO loss having approximately equal intensity. Secondly, electron autodetachment takes place in multiply charged anionic clusters, but only after elimination of a large proportion of their carbonyl ligands. Thirdly, clusters with open metal cores do not undergo CO loss in an orderly fashion, but certain peaks are considerably less intense. The appearance of these low-intensity peaks is believed to signify polyhedral core rearrangements, with open clusters folding to form more compact geometries. In some cases, the gas-phase transformations observed by EDESI-MS mirror those that are known to take place in solution. PMID- 12584711 TI - A computational study of regioselectivity in a cyclodextrin-mediated Diels-Alder reaction: revelation of site selectivity and the importance of shallow binding and multiple binding modes. AB - The use of a cyclodextrin.Diels-Alder transition structure complex (CD.TS) as a model in molecular dynamics simulations has enabled us to gain insight into the controlling factors in the cyclodextrin-mediated Diels-Alder reaction of methyl-p benzoquinone with isoprene. MD simulations were carried out with multiple binding configurations of the CD.TS (TS=meta-TS, para-TS) complexes at the top and bottom rims of beta-CD. We discovered that i) only shallow binding with the CD is necessary for the regioselectivity, and multiple binding geometries are possible; ii) the narrow bottom rim, with the primary hydroxyl groups, of the CD binds both regio-TSs better than at the wider top rim (secondary hydroxyl groups), which was unexpected from the perspective of shape complementarity that governs the stability of most CD.guest complexes. Overall, the bottom rim of the CD exhibits higher regioisomer discrimination for the meta-TS; iii) structural clustering analyses of the CD.TS configurations (sampled during MD simulations) have enabled us to evaluate the binding energies of the different binding configurations. The result indicates that there is a direct correlation between meta-product selectivity and a higher number of binding configurations favoring the formation of the CD.meta-TS complex. The main forces of stabilization in the CD.TS complexes are the van der Waals interactions when the TS is bound at the top rim. At the bottom rim, closer contacts between polar functional groups of the TS and CD have increased the importance of electrostatic interactions. We found that van der Waals, solvation, and torsional forces are less favorable for complexation at the bottom rim; however, this is compensated by large favorable electrostatic interactions. With insights obtained from the study of CD.TS complexes and MD simulations of the modified heptakis-[6-O-(2-hydroxy)propyl]-beta-CD, we were able to explain why a low selectivity was observed when the Diels-Alder reaction was carried out in this modified CD. Two types of search method [Monte Carlo and multiple minimum (MCMM) and molecular dynamics (MD)] to explore and evaluate the different possible binding geometries of the TS within beta-CD, were discussed. PMID- 12584712 TI - Neutral and ionic hydrogen bonding in Schiff bases. AB - Low-temperature, high-resolution X-ray studies of charge distributions in the three Schiff bases, the dianil of 2-hydroxy-5-methylisophthaldehyde, 3,5-dinitro N-salicylidenoethylamine and 3-nitro-N-salicylidenocyclohexylamine, have been carried out. These structures exhibit interesting weak interactions, including two extreme cases of intramolecular hydrogen bonds that are ionic N(+)-H...O- and neutral O-H...N in nature. These two types of hydrogen bond reflect differences in geometrical parameters and electron density distribution. At the level of geometry, the neutral O-H...N hydrogen bond is accompanied by an increase in the length of the C(1)-O(1) bond, opening of the ipso-C(1) angle, elongation of the aromatic C-C bonds, shortening of the C(7)-N(2) bond and increased length of the C(1)-C(7) bond, relative to the ionic hydrogen bond type. According to the geometrical and critical point parameters, the neutral O-H...N hydrogen bond seems to be stronger than the ionic ones. There are also differences between charge density parameters of the aromatic rings consistent with the neutral hydrogen bond being stronger than the ionic ones, with a concomitant reduction in the aromaticity of the ring. Compounds with the ionic hydrogen bonds show a larger double-bond character in the C-O bond than appears in the compound containing a neutral hydrogen bond; this suggests that the electronic structure of the former pair of compounds includes a contribution from a zwitterionic canonical form. Furthermore, in the case of ionic hydrogen bonds, the corresponding interaction lines appear to be curved in the vicinity of the hydrogen atoms. In the 3-nitro-N-salicylidenocyclohexylamine crystal there exists, in addition to the intramolecular hydrogen bond, a pair of intermolecular O...H interactions in a centrosymmetric dimer unit. PMID- 12584713 TI - Re-based heterogeneous catalysts for olefin metathesis prepared by surface organometallic chemistry: reactivity and selectivity. AB - Herein we describe the catalytic activity of 1, a well-defined Re alkylidene complex supported silica, in the reaction of olefin metathesis. This system is highly active for terminal and internal olefins with initial rates up to 0.7 mol per mol Re per s. It also catalyses the self-metathesis of methyl oleate (MO) without the need of co-catalysts. The turnover numbers can reach up to 900 for MO, which is unprecedented for a heterogeneous Re-based catalyst. Moreover the use of silica as a support can bring major advantages, such as the possibility to use branched olefins like isobutene, which are usually incompatible with alumina based supports; therefore, the formation of isoamylene from the cross-metathesis of propene and isobutene can be performed. All these results are in sharp contrast to what has been found for other silica- or alumina-supported rhenium oxide systems, which are either completely inactive (silica system) or typically need co-catalysts when functionalised olefins are used. Finally the initiation step corresponds to a cross-metathesis reaction to give a 3:1 mixture of 3,3 dimethylbutene and trans-4,4-dimethylpent-2-ene, and make this catalyst the first generation of well-defined Re-based heterogeneous catalysts. PMID- 12584714 TI - Combinatorial library of low molecular-weight organo- and hydrogelators based on glycosylated amino acid derivatives by solid-phase synthesis. AB - A combinatorial approach for the synthesis of supramolecular gelators as new organic materials is described herein. In the course of the development of a convenient and flexible solid-phase synthesis of the artificial glycolipids, some of these compounds were accidentally found to act as low molecular-weight gelators toward organic solvents. Using this combinatorial solid-phase synthesis of glycosylated amino acetates, screening and optimization of low molecular weight organo/hydro-gelators were efficiently carried out. We found that an N acetyl-galactosamine-appended amino acid ester (GalNAc-aa) efficiently gelates a broad spectrum of organic solvents. More interestingly, some GalNAc-aa derivatives displayed an excellent hydrogelation capability. Transmission electron microscopy, scanning electron microscopy, confocal laser scanning microscopy, and FT-IR were used for characterization of the gel structure. It is indicated that supramolecular fibers supported by strong hydrogen-bonding networks are entangled so that the resulting spaces can immobilize a number of solvent molecules effectively. In addition, the supramolecular hydrogel consisting of GalNAc-suc-glu(O-methyl-cyc-pentyl)(2) is stable even under high salt concentrations probably due to its nonionic character and as a result, a native protein is successfully entrapped in the gel matrix without denaturation. PMID- 12584715 TI - Preorganization and reorganization as related factors in enzyme catalysis: the chorismate mutase case. AB - In this paper a deeper insight into the chorismate-to prephenate-rearrangement, catalyzed by Bacillus subtilis chorismate mutase, is provided by means of a combination of statistical quantum mechanics/molecular mechanics simulation methods and hybrid potential energy surface exploration techniques. The main aim of this work is to present an estimation of the preorganization and reorganization terms of the enzyme catalytic rate enhancement. To analyze the first of these, we have studied different conformational equilibria of chorismate in aqueous solution and in the enzyme active site. Our conclusion is that chorismate mutase preferentially binds the reactive conformer of the substrate- that presenting a structure similar to the transition state of the reaction to be catalyzed--with shorter distances between the carbon atoms to be bonded and more diaxial character. With respect to the reorganization effect, an energy decomposition analysis of the potential energies of the reactive reactant and of the reaction transition state in aqueous solution and in the enzyme shows that the enzyme structure is better adapted to the transition structure. This means not only a more negative electrostatic interaction energy with the transition state but also a low enzyme deformation contribution to the energy barrier. Our calculations reveal that the structure of the enzyme is responsible for stabilizing the transition state structure of the reaction, with concomitant selection of the reactive form of the reactants. This is, the same enzymatic pattern that stabilizes the transition structure also promotes those reactant structures closer to the transition structure (i.e., the reactive reactants). In fact, both reorganization and preorganization effects have to be considered as the two faces of the same coin, having a common origin in the effect of the enzyme structure on the energy surface of the substrate. PMID- 12584716 TI - Towards a universal polymer backbone: design and synthesis of polymeric scaffolds containing terminal hydrogen-bonding recognition motifs at each repeating unit. AB - Polymers containing terminal hydrogen-bonding recognition motifs based on diaminotriazine and diaminopyridine groups in their side chains for the self assembly of appropriate receptors have been prepared by ring-opening metathesis polymerization (ROMP) of norbornenes. A new synthetic method for the preparation of norbornene monomers based on pure alkyl spacers is introduced. These monomers show unprecedented high reactivity using ROMP. To suppress self-association of diaminotriazine-based polymers, polymerizations were run in presence of N butylthymine. The butylthymine acts as a protecting group via self-assembly onto the hydrogen-bonding sites of the polymeric scaffold, thereby solubilizing the polymer. Diaminopyridine monomers do not require the presence of a protecting group due to their low propensity to dimerize. In addition, they exhibit a high affinity for hydrogen-bonded receptors on both monomeric and polymeric level. These polymers present our first building blocks towards the design and synthesis of a "universal polymer scaffold". PMID- 12584717 TI - Evidence for the intermediacy of arylbenzylnitrenium ions in the thermal rearrangement of isoxazolidines derived from C,N-diarylnitrones and 2-morpholin-4 yl-acrylonitrile. AB - In contrast to the diaryl, dialkyl, alkylaryl, and parent series, nothing is known about the generation and chemical behavior of arylbenzylnitrenium ions. Herein, we report that these species can be generated by a process involving an unprecedented thermal rearrangement of isoxazolidines derived from C,N diarylnitrones and 2-morpholin-4-yl-acrylonitrile. The products from these reactions are dramatically dependent upon the nature of the nitrone. Most of the observed chemistry originates from the singlet state. PMID- 12584718 TI - Conformers of gaseous proline. AB - Accurate geometries, relative energies, rotational and quartic centrifugal distortion constants, dipole moments, harmonic vibrational frequencies, and infrared intensities were determined from ab initio electronic structure calculations for eighteen conformers of the neutral form of the amino acid L proline. Only four conformers have notable population at low and moderate temperature. The second most stable conformer is only 2+/-2 kJ mol(-1) above the global minimum, while the third and fourth conformers are nearly degenerate and have an excess energy of 7+/-2 kJ mol(-1) relative to the global minimum. All four conformers have one hydrogen bond: N.HO in the lower energy pair of conformers, and NH.O in the higher energy pair of conformers. The conformer pairs differ only in their ring puckering. The relative energies of the conformers include corrections for valence electron correlation, extrapolated to the complete basis set limit, as well as core correlation and relativistic effects. Structural features of the pyrrolidine ring of proline are discussed by using the concept of pseudorotation. The accurate rotational and quartic centrifugal distortion constants as well as the vibrational frequencies and infrared intensities should aid identification and characterization of the conformers of L proline by rotational and vibrational spectroscopy, respectively. Bonding features of L-proline, especially intramolecular hydrogen bonds, were investigated by the atoms-in-molecules (AIM) technique. PMID- 12584720 TI - Interactions among memory-related centers in the brain. AB - The structures associated with learning and memory have been widely studied for over 100 years. The idea of the famous neuropsychologist K.S. Lashley, that learning and memory are stored diffusely in the brain, dominated neuroscience in the early half of Twentieth Century. Since Scoville reported in 1957 a persistent impairment of recent memory caused by bilateral medial temporal lobe resection in a patient, the concept that different brain structures play different roles in learning and memory has been established, but the structures were thought to work separately. The connections and functional influences between hippocampus and prefrontal cortex, thalamus and hippocampus, prefrontal cortex and thalamus, amygdala and hippocampus, basal nucleus of Meynert and medial temporal lobe system, and amygdala and thalamus were successively reported. The marginal division (MrD) is a pan-shaped structure consisting of spindle-shaped neurons at the caudal margin of the neostriatum in the mammalian brain. The MrD has been shown to contribute to associative learning and declarative memory by behavioral study in rats and by functional magnetic resonance image study in humans. Lesions in the MrD influenced the learning and memory function of the basal nucleus of Meynert and attenuated hippocampal long-term potentiation. The MrD is likely, based on its position, advanced development in higher mammalian brains, abundant and swift blood supply, and complex connections, to be an important subcortical memory center in the brain. The above-mentioned studies demonstrated that memory related centers could influence each other and play different roles. Therefore, we propose that there are very possibly hierachical memory centers in the brain. PMID- 12584721 TI - Differential expression of connexin 43 in the chick tangential vestibular nucleus. AB - The chick tangential nucleus is a major vestibular nucleus whose principal cells receive convergent inputs from primary vestibular and nonvestibular fibers and participate in the vestibular reflexes. During development, the principal cells gradually acquire the mature firing pattern in part by losing a specific potassium current around hatching (H). Here we focus on characterizing the expression of connexin 43 (Cx43), a gap junction protein found mainly between astrocytes in the mature brain. The astrocytic syncytium plays an important role in maintaining extracellular potassium ion balance in the brain. Accordingly, it is important to characterize the potential of this syncytium to communicate during the critical developmental age of hatching. Using fluorescence immunocytochemistry, we investigated whether Cx43 staining was concentrated in specific cellular compartments at H1 by applying well-known markers for astrocytes (glial fibrillary acidic protein; GFAP), oligodendrocytes (antimyelin), neurons (microtubule-associated protein 2), and synaptic terminals (synaptotagmin). GFAP-positive astrocytes and GFAP-negative nonneuronal cells around the principal cell bodies were labeled with Cx43, suggesting that Cx43 was expressed exclusively by nonneuronal cells near the neuronal elements. Next, the developmental pattern of expression of Cx43 was studied at embryonic day 16 (E16), H1, and H9. At E16, Cx43 was present weakly as random small clusters in the tangential nucleus, whereas, at H1, overall staining became localized, with increases in size, brightness, and number of immunostained clusters. Finally, at H9, Cx43 staining decreased, but cluster size and location remained unchanged. These results suggest that Cx43 is developmentally regulated with a peak at birth and is associated primarily with astrocytes and nonneuronal cells near the principal cell bodies. PMID- 12584722 TI - Intracellular fibroblast growth factor produces effects different from those of extracellular application on development of avian cochleovestibular ganglion cells in vitro. AB - In an avian coculture system, the neuronal precursors of the cochleovestibular ganglion typically migrated from the otocyst and differentiated in response to soluble fibroblast growth factor (FGF-2), which had free access to FGF receptors on the cell surface. Free FGF-2 switched cells from a proliferation mode to migration, accompanied by increases in process outgrowth, fasciculation, and polysialic acid expression. Microsphere-bound FGF-2 had some of the same effects, but in addition it increased proliferation and decreased fasciculation and polysialic acid. As shown by immunohistochemistry, FGF-2 that was bound to latex microspheres depleted the FGF surface receptor protein, which localized with the microspheres in the cytoplasm and nucleus. For microsphere-bound FGF-2, the surface receptor-mediated responses to FGF-2 appear to be limited and the door opened to another venue of intracellular events or an intracrine mechanism. PMID- 12584723 TI - Neurotrophins facilitate neuronal differentiation of cultured neural stem cells via induction of mRNA expression of basic helix-loop-helix transcription factors Mash1 and Math1. AB - Neurogenesis is promoted by basic helix-loop-helix (bHLH) transcription factors Mash1, Math1, or NeuroD but suppressed by another set, Hes1 and Hes5. It remains unknown what kinds of extracellular signals are involved in their regulation; therefore, the effects of neurotrophins on the expression of bHLH factors and neuronal differentiation were investigated by the use of cultured mouse neural stem cells. Each neurotrophin increased Mash1 and Math1 mRNAs of the stem cells growing in the presence of fibroblast growth factor-2 (FGF-2), but did not alter Hes1, Hes5, or NeuroD mRNA levels. Simultaneously, most of the cells expressed nestin but not microtubule-associated protein 2 (MAP2), and remained undifferentiated. FGF-2 removal from the medium reduced the levels of Hes1 and Hes5 mRNAs and increased those of Mash1, Math1, and NeuroD mRNAs, resulting in substantial neuronal differentiation. When the cells were pretreated with brain derived neurotrophic factor, a neurotrophin, FGF-2 removal enhanced earlier NeuroD expression and generated many more MAP2-positive cells. The high level of Mash1 and Math1 that had been elevated at FGF-2 withdrawal accelerated NeuroD expression in cooperation with the reduced Hes1 and Hes5 expression. Our present results suggest that neurotrophins stimulate neuronal differentiation by altering the balance of expression of various bHLH transcription factors. PMID- 12584725 TI - Thyroid hormones regulate the frequency of miniature end-plate currents in pre- and prometamorphic stages of the tadpole tail. AB - Thyroid hormones (THs), primarily 3,3',5-triiode-(L)-thyronine (T(3)), have been clearly established as natural inducers of apoptosis during metamorphosis of anuran embryos. We decided to use this phenomenon to test the hypothesis that, prior to genomic activation, T(3) has acute actions in the neuromuscular junction (NMJ) of the tail of amphibian embryos. We detected a dramatic increase in the production of miniature end-plate currents (MEPCs) 2-5 min after continuous application of T(3) (250 nM) using focal recordings under voltage clamp. Furthermore, this increase in the spontaneous release of neurotransmitter, evaluated by the MEPC frequency, was maintained for several hours. Reverse-T(3), the "inhibitory" form of THs, prevented this increase in MEPC frequency, suggesting that this is probably a highly specific action of T(3). In addition, the elevation in MEPC frequency induced by T(3) was unchanged in the presence or absence of extracellular calcium. The T(3)-mediated increase in MEPC frequency was blocked by niflumic acid, a nonsteroidal antinflammatory fenamate used to prevent the apoptotic volume decrease observed in many systems. The present study demonstrated that T(3) induces a remarkable nongenomic action in the NMJ of the tadpole tail at pre- and promatamorphic stages. PMID- 12584724 TI - Erythropoietin prevents early and late neuronal demise through modulation of Akt1 and induction of caspase 1, 3, and 8. AB - Erythropoietin (EPO) modulates primarily the proliferation of immature erythroid precursors, but little is known of the potential protective mechanisms of EPO in the central nervous system. We therefore examined the ability of EPO to modulate a series of death-related cellular pathways during anoxia and free radical induced neuronal degeneration. Neuronal injury was evaluated by trypan blue, DNA fragmentation, membrane phosphatidylserine exposure, protein kinase B phosphorylation, cysteine protease activity, mitochondrial membrane potential, and mitogen-activated protein (MAP) kinase phosphorylation. We demonstrate that constitutive neuronal EPO is insufficient to prevent cellular injury, but that signaling through the EPO receptor remains biologically responsive to exogenous EPO administration. Exogenous EPO is both necessary and sufficient to prevent acute genomic DNA destruction and subsequent phagocytosis through membrane PS exposure, because neuronal protection by EPO is completely abolished by co treatment with an anti-EPO neutralizing antibody. Through pathways that involve the initial activation of protein kinase B, EPO maintains mitochondrial membrane potential. Subsequently, EPO inhibits caspase 8-, caspase 1-, and caspase 3-like activities linked to cytochrome c release through mechanisms that are independent from the MAP kinase systems of p38 and JNK. Elucidating some of the novel neuroprotective pathways employed by EPO may further the development of new therapeutic strategies for neurodegenerative disorders. PMID- 12584726 TI - Oxidative neuronal death caused by glutamate uptake inhibition in cultured hippocampal neurons. AB - Glutamate transporters are coupled with cystine/glutamate antiporters to supply cystine as a component of glutathione, an important antioxidant. We sought evidence that L-trans-pyrrolidine-2,4-dicarboxylate (PDC) enhances glutamate induced neuronal damage not only via the N-methyl-D-aspartate (NMDA) receptor mediated pathway, but also through induction of oxidative stress. Cultured hippocampal cells were exposed to glutamate (100 microM) for 5 min, washed and incubated for 18 hr with PDC (200 microM). PDC, increasing the neuronal death to 147% of that induced by glutamate alone, depleted glutathione in the culture, and produced dichloro-dihydro-fluorescein-diacetate-positive reactive oxygen species in neurons. N-acetylcysteine (2 mM) not only reduced PDC-enhanced neuronal death but also recovered glutathione and abolished the reactive oxygen species in these neurons. Threo-beta-benzyloxyaspartate, another type of glutamate transporter inhibitor, also induced glutathione depletion in the glutamate-preloaded cells, suggesting the involvement of glutamate transporter blocking in glutathione depletion. The NMDA receptor antagonist MK-801, although partially effective in reducing PDC toxicity, slightly recovered glutathione level but did not reduce the reactive oxygen species even at a high concentration (100 microM). N acetylcysteine, dimethylsulfoxide, alpha-phenyl-N-butyl nitrone and glutathione ethylester prevented neuronal death enhanced by PDC, but superoxide dismutase and catalase did not. Our study provides evidence that the block of glutamate uptake by PDC exerts toxicity on glutamate-pretreated neurons not only through the accumulation of extracellular glutamate and subsequent activation of the NMDA receptor but also through depletion of glutathione and generation of reactive oxygen species. PMID- 12584727 TI - Pyrrolidine dithiocarbamate-induced neuronal cell death is mediated by Akt, casein kinase 2, c-Jun N-terminal kinase, and IkappaB kinase in embryonic hippocampal progenitor cells. AB - Pyrrolidine dithiocarbamate (PDTC) is known to induce cell death by the stimulation of intracellular zinc transport and subsequent modulation of nuclear factor-kappaB (NF-kappaB) activity. Zinc is a signaling messenger that is released by neuronal activity at many central excitatory synapses. Excessive synaptic release of zinc followed by entry into vulnerable neurons contributes to severe neuronal cell death. In the present study, we explored how PDTC modulates intracellular signal transduction pathways, leading to neuronal cell death. The exposure of immortalized embryonic hippocampal cells (H19-7) to PDTC within the range of 1-100 microM caused cell death in a dose-dependent manner. During the cell death, NF-kappaB activity increased in response to PDTC, and this activity corresponded well with the increase of intracellular free zinc levels, implying that the activation of NF-kappaB transmits the cell death signals of PDTC. Furthermore, PDTC caused the activation of IkappaB kinase (IKK), casein kinase 2 (CK2), phosphatidylinositol 3-kinase (PI-3K), and Akt, as well as mitogen activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), but not p38 kinase. The blockade of PI 3K, JNK, and CK2 pathways resulted in a remarkable suppression of PDTC-induced cell death and also the activation of IKK, which subsequently led to a decrease of IkappaB phosphorylation. Although the overexpression of dominant-negative SEK in a transient manner did not inhibit the activation of Akt by PDTC, the transfection of kinase-inactive Akt mutants did cause a remarkable blockade of JNK activation, implying that Akt is present upstream of JNK in the PDTC signaling pathways. Moreover, whereas selective CK2 inhibitors suppressed PDTC induced JNK activation, the inhibition of JNK did not affect CK2 activity, suggesting that CK2 is directly related to the regulation of cell viability by PDTC and that the CK2-JNK pathway could be a downstream target of PDTC. Taken together, our results suggest that PDTC-mediated accumulation of intracellular zinc ions may affect cell viability by modulating several intracellular signaling pathways in neuronal hippocampal progenitor cells. PMID- 12584728 TI - Organic cation transporter capable of transporting serotonin is up-regulated in serotonin transporter-deficient mice. AB - The serotonin (5HT) transporter (5HTT) regulates serotonergic neurotransmission by mediating the reuptake of 5HT from the synaptic cleft. Although lacking the high affinity and selectivity of the 5HTT, the brain expresses a large number of other transporters, including the polyspecific organic cation transporters (OCTs). OCT1 and OCT3, members of the potential-sensitive organic cation transporter gene family, physiologically transport a wide spectrum of organic cations. In addition, both transporters mediate low-affinity 5HT transport and, therefore, may participate in the clearance of excessive 5HT. Because concentrations of extracellular 5HT are increased in the brain of 5HTT-deficient mice, they are a model for investigating the role of OCTs in 5HT system homeostasis. Here, we analyzed OCT1 and OCT3 gene expression in the brain of 5HTT knockout mice by semiquantitative competitive polymerase chain reaction and in situ hybridization. We demonstrate that, in 5HTT-deficient mice, OCT3 mRNA concentrations were significantly increased in the hippocampus, but not in other brain regions, including cortex, striatum, cerebellum, and brainstem. In contrast, no difference in OCT1 expression was detected between 5HTT knockout and control mice. Up-regulation of OCT3 expression and enhanced low-affinity 5HT uptake may limit the adverse effects of elevated extracellular 5HT and may play a critical role in maintaining 5HT-dependent functions of the hippocampus in the absence of 5HTT. PMID- 12584729 TI - Altered expression of randomly selected genes in mouse hippocampus after traumatic brain injury. AB - Using a cDNA microarray method, we analyzed gene expression profiles in mouse hippocampus after traumatic brain injury (TBI). Of 6,400 randomly selected arrayed genes and expressed sequence tags from a mouse cDNA library, 253 were found to be differentially expressed (106 increased and 147 decreased). Genes involved in cell homeostasis and calcium signaling were primarily up-regulated while those encoding mitochondrial enzymes, metabolic molecules, and structural proteins were predominantly down-regulated. Equal numbers of genes related to inflammatory reactions showed increased or decreased expression. Importantly, a large proportion of the dysregulated genes we identified have not been reported as differentially expressed in TBI models. Semiquantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analyses of representative genes confirmed the validity of the corresponding microarray findings. Thus, our microarray-based evaluation of gene expression in traumatically injured hippocampus identified both known and novel genes that respond to TBI. Further investigation of these candidate molecules may suggest new ways to attenuate the traumatic effects of brain injury. PMID- 12584730 TI - Tumor necrosis factor-alpha regulation of insulin-like growth factor-I, type 1 IGF receptor, and IGF binding protein expression in cerebellum of transgenic mice. AB - Tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine, has been implicated in the pathogenesis of several disorders and injuries in the central nervous system (CNS). Unlike IGF-I, which promotes CNS growth, TNF-alpha causes brain growth retardation and neural damage. Recently TNF-alpha has been shown to inhibit IGF-I signaling and actions in non-neural tissue. To investigate whether TNF-alpha deleteriously influences brain growth by altering the IGF-I system in vivo, we examined the expression of IGF-I, the type 1 IGF receptor (IGF1R) and IGF binding proteins (IGFBPs) in the brain of transgenic (Tg) mice with murine TNF-alpha overexpression. We show that overexpression of TNF-alpha reduces the weights of whole brain and all brain regions examined during development. In adult TNF-alpha Tg mice, cerebellum (CB) exhibited the greatest reduction in weight among the five brain regions examined, being approximately 77% of that in wild-type (WT) mice. IGF-I abundance was decreased in the CB, as well as in cerebral cortex and diencephalon, of TNF-alpha Tg mice. When compared to those in WT mice, CB IGF-I abundance in Tg mice was reduced by approximately 35%, approximately 45%, and approximately 40% at 2, 6, and 9 weeks of age, respectively. Of the IGFBPs studied the abundance of IGFBP-3 and IGFBP-4 was increased by 2-3.7-fold, and the abundance of IGFBP-5 was decreased by approximately 3-fold (as judged by Western immunoblot analysis). Histological analysis and immunocytochemical staining confirmed that TNF-alpha specifically increases IGFBP-3 and IGFBP-4 immunoreactivity, as well as that of the IGF1R, in radial glial and Purkinje cells. In addition, TNF-alpha alters CB cytoarchitecture, apparently by influencing granule cell migration. Our data indicate that TNF-alpha alters the expression of IGF-I system proteins in vivo, and suggest that altered expression of IGF-I system proteins may in part explain TNF-alpha deleterious actions on brain growth. PMID- 12584731 TI - Kif1Bbeta isoform is enriched in motor neurons but does not change in a mouse model of amyotrophic lateral sclerosis. AB - The kinesin superfamily motor protein Kif1B is expressed in two isoforms, Kif1Balpha and Kif1Bbeta, with distinct cargo-binding domains. We examined the mRNA distribution of the two isoforms in adjacent sections of brain and spinal cord of adult mice using in situ hybridization analysis. Kif1Bbeta mRNA is enriched in several regions of brain and spinal cord. Its levels are four to five times higher than that of the alpha isoform, which was barely detectable. The highest mRNA levels of Kif1Bbeta were found in the cortex, hippocampus, cerebellum and the grey matter of the spinal cord. At the cellular level the highest signal was found in motor neurons in the motor nuclei of medulla oblongata and the ventral horn of spinal cord. Because expression of other Kif genes is altered in amyotrophic lateral sclerosis (ALS) models, we examined the expression level of Kif1Bbeta mRNA in the spinal cord of transgenic mice carrying the SOD1G93A mutation, a model of familial ALS, at presymptomatic and early stages of the disease. No changes were observed in Kif1Bbeta mRNA in motor neurons or in other regions of the spinal cord. These findings indicate that Kif1Balpha, which modulates the transport of mitochondria, may play a major role in tissues other than the central nervous system. Instead Kif1Bbeta, responsible for the transport of synaptic vesicle precursors, seems to play an important role in the nervous system, particularly in the lower motor neurons. The absence of changes of Kif1Bbeta mRNA in transgenic SOD1G93A mice suggests that other molecular mechanisms may play a role in the disruption of axonal transport occurring in the motor neurons of these mice. PMID- 12584732 TI - Chlamydia pneumoniae infection promotes the transmigration of monocytes through human brain endothelial cells. AB - We have investigated the effects of Chlamydia pneumoniae on human brain endothelial cells (HBMECs) and human monocytes as a mechanism for breaching the blood-brain barrier (BBB) in Alzheimer's disease (AD). HBMECs and peripheral blood monocytes may be key components in controlling the entry of C. pneumoniae into the human brain. Our results indicate that C. pneumoniae infects blood vessels and monocytes in AD brain tissues compared with normal brain tissue. C. pneumoniae infection stimulates transendothelial entry of monocytes through HBMECs. This entry is facilitated by the up-regulation of VCAM-1 and ICAM-1 on HBMECs and a corresponding increase of LFA-1, VLA-4, and MAC-1 on monocytes. C. pneumoniae infection in HBMECs and THP-1 monocytes up-regulates monocyte transmigration threefold in an in vitro brain endothelial monolayer. In this way, C. pneumoniae infection in these cell types may contribute to increased monocyte migration and promote inflammation within the CNS resulting from infection at the level of the vasculature. Thus, infection at the level of the vasculature may be a key initiating factor in the pathogenesis of neurodegenerative diseases such as sporadic AD. PMID- 12584733 TI - New component of the limbic system: Marginal division of the neostriatum that links the limbic system to the basal nucleus of Meynert. AB - The limbic system refers to a group of connected neural regions that are associated with motivation, learning, and memory. The marginal division (MrD) is a zone located at the caudal border of the neostriatum in mammalian brains that has been shown to be involved in learning and memory. In a previous study, c-fos expression showed functional connections between the MrD, basal nucleus of Meynert (NBM) and limbic system (Shu et al., 1988a, 1999). In the present study, to explore the relationship between these regions, the expression of limbic system-associated membrane protein (LAMP) was investigated using molecular and immunohistochemical methods. Synaptic and functional connections between the MrD and the NBM were studied also using tract tracing, electron microscopic and behavioral methods. LAMP is thought to be a marker of the limbic system and expression of LAMP protein and mRNA was observed in both the MrD and the limbic system. From such results, it is concluded that the MrD is a new component of the limbic system. Fibers from the MrD were observed projecting and synapsing on cholinergic neurons of the NBM. As reduction of learning and memory was induced by lesioning the projection from the MrD to the NBM, it would seem that the MrD modulates the learning and memory function of the NBM. In conclusion, the results of these studies suggest that the MrD is a new component of the limbic system, and there are functional and structural connections between the MrD, NBM and limbic system. The MrD seems to act as a link between the limbic system and the NBM, and plays a role in learning and memory. PMID- 12584734 TI - Macrophages in tumor-draining lymph node with different characteristics induce T cell apoptosis in patients with advanced stage-gastric cancer. AB - A host's immune-defense system is suppressive by many factors in patients with cancer. We have previously shown one possible mechanism behind the T-cell dysfunction, whereby H(2)O(2) secreted from macrophages in tumor-draining lymph node (MTDL) induced T-cell dysfunction with down-regulation of TCR zeta molecules. In the present study, we analyzed how MTDL affect T cells, with a particular focus on T-cell apoptosis, by co-culturing MTDL with autologous peripheral blood T cells in gastric cancer. Moreover, we characterized the MTDL according to surface marker, oxygen-burst capacity and intracellular cytokine status. T-cell apoptosis was significantly induced in comparison to T-cell alone control in patients with advanced disease, concomitant to the elevated caspase activity and following impaired T-cell function. In patients with early disease, no significant difference was seen in the proportions of T cells that underwent apoptosis between T cells plus MTDL and T cells alone. Moreover, the addition of a selective scavenger of H(2)O(2), catalase inhibited the apoptosis of T cells co cultured with MTDL in patients with advanced disease. In the characterization of MTDL, the production of H(2)O(2) in MTDL from advanced disease was significantly higher than that in early disease. The amounts of intracellular IL-10 and IL-12 in MTDL in advanced disease were significantly higher than those in early disease. These results indicated that MTDL induced apoptosis of autologous T cells and this T-cell dysfunction was mediated by H(2)O(2) derived from MTDL. Furthermore, the characteristics of MTDL including the capacity of oxygen-burst and intracellular cytokine production were different depending on the disease progression. PMID- 12584736 TI - Enhancement of TRAIL/Apo2L-mediated apoptosis by adriamycin through inducing DR4 and DR5 in renal cell carcinoma cells. AB - Renal cell carcinoma (RCC) is one of the most drug-resistant malignancies in humans. We show that adriamycin (ADR) and TNF-related apoptosis-inducing ligand (TRAIL)/Apo2L have a synergistic cytotoxic effect against RCC cells. This synergistic cytotoxicity was obtained in ACHN, A704, Caki-1 and Caki-2 human RCC cell lines and freshly derived RCC cells from 6 patients. This synergistic effect, however, was not achieved in 5 samples of freshly isolated normal kidney cells. We further explored the mechanisms underlying this synergistic effect and found that the synergistic cytotoxicity of TRAIL/Apo2L and ADR was realized by inducing apoptosis. Sequential treatment with ADR followed by TRAIL/Apo2L induced significantly more cytotoxicity than the reverse treatment. ADR increased the expression of DR4 and DR5 in RCC cells, but not in the normal kidney cells. Furthermore, the synergistic cytotoxicity was significantly inhibited by DR4:Fc and DR5:Fc fusion proteins, which inhibit TRAIL/Apo2L-mediated apoptosis. In addition, caspase activity assays and treatment of caspase inhibitors demonstrated that the combination treatment with ADR and TRAIL/Apo2L activated caspase cascade, including caspase-9, -8, -6 and -3, which were the downstream molecules of death receptors. These findings indicate that ADR sensitizes RCC cells to TRAIL/Apo2L-mediated apoptosis through induction of DR4 and DR5, suggesting that the combination therapy of TRAIL/Apo2L and ADR might be effective for RCC therapy. PMID- 12584735 TI - Role of IRF-1 and caspase-7 in IFN-gamma enhancement of Fas-mediated apoptosis in ACHN renal cell carcinoma cells. AB - Caspases exist as zymogens, and are activated by various extracellular stimuli, leading to apoptosis. One such stimulus is Fas/CD95, a member of the tumor necrosis factor receptor family, providing one means of cytotoxic T lymphocyte (CTL)-mediated cell lysis. Clinical evidence has shown that administration of cytokine leads to regression in selected patients with renal cell carcinomas (RCCs). Interferon-gamma (IFN-gamma) indicates its contribution to anti-tumor activity of immune cells. IFN-gamma elicits its effect through the transcription factor signal transducer and activator of transcription-1 (STAT-1), and through interferon regulatory factor-1 (IRF-1), one of the target genes of STAT-1. Our previous study demonstrated an increase in the susceptibility of ACHN cells, established from RCC, to Fas-mediated apoptosis by IFN-gamma, and the inhibition of this effect by the caspase-3 and -7 inhibitor, DEVD-CHO. We demonstrated the following phenomena in IFN-gamma-treated ACHN cells: 1) enhanced transcription of caspase-1, 3 and 7 mRNAs without any change in cleavage of their substrates; 2) increased cleavage DEVD (specific for caspase-3 and 7), but not YVAD (for caspase 1) or DMQD (for caspase-3), after anti-Fas/CD95 MAb treatment; 3) activation of the STAT-1 and IRF-1 pathway; and 4) partial abrogation of the IFN-gamma-induced increase in Fas-mediated apoptosis by antisense IRF-1 oligodeoxynucleotide. These results suggest that IRF-1 plays a pivotal role in the IFN-gamma-mediated enhancement of Fas/CD95-mediated apoptosis, through regulation of DEVD-CHO sensitive caspases, most likely caspase-7. PMID- 12584737 TI - The synthetic retinoid RO 13-6307 induces neuroblastoma differentiation in vitro and inhibits neuroblastoma tumour growth in vivo. AB - Retinoids modulate cell proliferation, differentiation and apoptosis in a variety of tumour cells including leukaemia and neuroblastoma, a childhood tumour of the sympathetic nervous system. 13-cis retinoic acid is in clinical use against minimal residual disease in neuroblastoma, where the effect seems to depend on dose, scheduling and tumour mass. Novel retinoids are searched for, to improve potency and lower toxicity. We investigated the effect of the synthetic retinoid Ro 13-6307 on neuroblastoma growth in vitro on SK-N-BE(2) and SH-SY5Y cells. Furthermore, effects on tumour growth and the toxicity profile were investigated in a rat xenograft model. Effects of Ro 13-6307 were compared to 13-cis RA (retinoic acid) in vitro and in vivo. Neuroblastoma cells treated with 1 microM Ro 13-6307 exhibited neuronal differentiation, decreased proliferation and accumulation of cells in G1 phase in at least the same magnitude as 5 microM 13 cis RA. No apoptosis was detected in vitro. Treatment of nude rats with neuroblastoma using Ro 13-6307, 0.12 mg p.o. daily, decreased neuroblastoma growth in vivo, in terms of tumour volume during treatment and tumour weight at sacrifice (p < 0.05). In contrast, Ro 13-6307, 0.08 mg p.o. daily, resulted in no significant reduction in tumour growth. All rats treated with Ro 13-6307 gained less weight than control rats, but they exhibited no other signs of toxicity. The toxicity profile of Ro 13-6307 was similar to what we found with 13-cis RA. Our preclinical results suggest that Ro 13-6307 may be a candidate retinoid for clinical oral therapy of neuroblastoma in children. PMID- 12584738 TI - C-Raf controlled pathways in the protection of tumor cells from apoptosis. AB - The Raf serine-threonine kinase is upregulated in many human tumors and plays a pivotal role in tumor cell proliferation and survival. Abrogation of c-Raf expression by specific antisense oligonucleotides (Raf-AS-ODN) efficiently blocks tumor cell growth and induces apoptosis in human cancer cells. The signaling pathways and molecular mechanisms c-Raf utilizes to mediate the survival of tumor cells are, however, not well understood. Here we show that apoptosis triggered by Raf depletion cannot be overcome by ectopic Bcl-2 expression and occurs in the absence of cytochrome c release, arguing against a direct impact of c-Raf on mitochondrial pathways of apoptosis regulation. We also show that c-Raf depletion leads to a clearly decreased expression of different epidermal growth factor (EGF) receptor ligands, suggesting that the autocrine stimulation of an EGF receptor-mediated survival pathway might be involved in the blockade of tumor cell apoptotis by c-Raf. PMID- 12584739 TI - Methylation pattern of HLTF gene in digestive tract cancers. AB - Recently, the loss of helicase-like transcription factor (HLTF) gene expression accompanied by HLTF promoter methylation was identified in colon cancers. We examined HLTF methylation in colorectal, gastric and esophageal carcinomas. Twenty-five of 76 colorectal cancers (33%), 11 of 65 gastric cancers (17%), and 1 of 40 esophageal cancers (3%) demonstrated abnormal methylation of the HLTF promoter. Abnormal methylation was found in the tumors of patients at all clinical stages, suggesting that these cancers could be methylated from an early stage. These results suggested that HLTF might play a variety of roles depending on the tissue type and could be inactivated at an early stage in the tumorigenic pathway. PMID- 12584740 TI - Treatment of colon and breast carcinoma cells with 5-fluorouracil enhances expression of carcinoembryonic antigen and susceptibility to HLA-A(*)02.01 restricted, CEA-peptide-specific cytotoxic T cells in vitro. AB - Cancer vaccines directed against tumor associate antigen (TAA) have produced encouraging results in preclinical models but not in cancer patients. A major limitation of this strategy is the relative degree of tolerance to these antigens and the low and heterogeneous tumor cell expression of TAA and major histocompatibility complex (MHC). Previous studies have shown that 5-fluorouracil (5-FU) can upregulate the expression of membrane-associated carcino-embryonic antigen (CEA), and MHC molecules in colon and breast carcinoma cell lines. We have investigated whether this drug can also enhance their sensitivity to the lytic effects of CEA-peptide specific Cytotoxic T cell lymphocytes (CTL). The CEA peptide-specific CTLs generated in our laboratory from normal HLA-A(*)02.01(+) donor PBMCs, were able to kill HLA-A(*)02.01(+)/CEA(+) breast (MCF-7-T103) and colon (HLA-A(*)02.01 gene-transfected HT-29 and C22.20) carcinoma cells in HLA A(*)02.01 restricted manner. The treatment of target cells with 5-FU, enhanced their CEA expression and susceptibility to CTL-mediated lysis. Cold competition assays confirmed these results, thus supporting the hypothesis that immune target cell lysis and 5-FU mediated enhancement were dependent on CEA peptide presentation by cancer cells. 5-FU treatment of functionally "mature" CTL after in vitro expansion, did not reduce their cytolytic activity against MT-2 target cells but, when the anti-metabolite was added during the immune-sensitization phase, CTL generation was significantly inhibited. These results provide a rationale for investigating a possible new role of 5-FU as an immuno targeting amplifier agent in breast and colorectal cancer patients immunized with CEA directed cancer vaccines. PMID- 12584741 TI - SMAD7 is a prognostic marker in patients with colorectal cancer. AB - Chromosomal region 18q21 is frequently deleted in colorectal cancer (CRC) and is associated with poor prognosis. Potential tumor suppressor mechanisms altered by 18q21 deletion include mediation of TGFbeta signaling by SMADs. Following the definition of SMAD4 deletion as a negative predictive marker for chemotherapy benefit in patients with CRC, we aimed to evaluate the clinical relevance of the deletion of other SMAD genes clustered in this region: SMAD2 and SMAD7 in 264 CRC biopsies from a previous clinical study. In contrast to SMAD2 deletion, for which no clinical relevance was observed, hazard ratios (HR) in a multivariate analysis associated with SMAD7 deletion [overall survival (OS): HR = 0.43, p = 0.0012; disease-free survival (DFS): HR = 0.50, p = 0.0033] indicated a favorable outcome for these patients. In addition, SMAD7 duplication had a hazardous effect on survival [OS: HR = 2.10, p = 0.020; DFS: HR = 2.06, p = 0.015]. Moreover, the HRs associated with one additional copy of SMAD7 were 1.76, p = 0.00024 [OS] and 1.64, p = 0.00048 [DFS] respectively, showing a graded effect of SMAD7 on patient outcome depending on gene copy number that suggests a dose-and-effect basis. Since SMAD7 blocks TGFbeta signaling, these data are consistent with the loss of SMAD7 rendering carcinoma cells more sensitive to cell growth arrest/apoptotic effect of TGFbeta, whereas gain of SMAD7 function might result in TGFbeta resistance, thereby emphasizing the role of TGFbeta in tumor suppression. PMID- 12584742 TI - HSD17B1 and CYP17 polymorphisms and breast cancer risk among Chinese women in Singapore. AB - Reasons for the recent trend of increasing breast cancer incidence among Chinese and other Asian women are not well understood. Endogenous estrogen levels are strongly associated with breast cancer risk and its determinants include both genetic and lifestyle factors. We conducted a nested case-control study to investigate, within the Singapore Chinese Health Study Cohort, the relationships between polymorphisms in 2 genes involved in estrogen metabolism, CYP17 and HSD17B1, and the risk of breast cancer. For this analysis, 188 incident breast cancer cases and 671 female cohort control subjects were compared. When the HSD17B1 A allele was considered as the "putative high-risk" allele, there was a modest increased risk (adjusted relative risk, RR=1.37, 95% CI=0.90-2.07 for HSD17B1 AA vs. other); this association was statistically significant in analysis restricted to postmenopausal women (RR=1.86, 95% CI=1.14-3.03). There was no significant association between the CYP17 MspAI polymorphism and risk in all subjects (RR=1.06, 95% CI=0.65-1.74 for CYP17 A2A2 vs. CYP17 A1A1) or in postmenopausal women only. When we evaluated breast cancer risk in relation to the joint stratification of CYP17 and HSD17B1 genotypes and according to the combined number of putative high-risk alleles (range, 0-4), we observed an elevated joint effect of the CYP17 and HSD17B1 genes on risk. Women who possessed all 4 putative high-risk alleles of both genes (CYP17 A2A2 and HSD17B1 AA) vs. less displayed a nearly 2-fold increased risk (RR=1.83, 95% CI=0.97-3.44); this finding was statistically significant in postmenopausal women (RR=2.31, 95% CI=1.07-4.98). Risk of breast cancer was similar among women possessing the other genotypes (i.e., less than 4 putative high-risk alleles in the joint CYP17/HSD17B1 genotypes). In addition, the significant increased risk of breast cancer associated with nulliparity or late age at first live birth (age 31 years or older) was largely limited to women with the high-risk CYP17 A1A2/A2A2 or HSD17B1 AA genotypes (RR=2.41, 95% CI=1.56-3.72; RR=4.39, 95% CI=1.71-11.30, respectively). The latter gene-parity effects were especially pronounced in postmenopausal women. PMID- 12584743 TI - Prostate cancer and exposure to pesticides in agricultural settings. AB - Our study evaluates the association between prostate cancer and exposure to pesticides in agricultural settings in Italy. The data were derived from a hospital-based multi-site case-control study carried out in 5 rural areas between 1990-92. In our study, 124 new cases of prostate cancer were ascertained and interviewed, along with 659 cancer controls. A team of agronomists assessed past exposure to pesticides by using a checklist of 100 chemical families and 217 compounds applied from 1950-85 in the areas considered. The association between prostate cancer and different occupational risk factors was measured by maximum likelihood estimation of the odds ratio, controlling for potential confounders. "Ever been employed in agriculture" was associated with a 40% increased risk (OR = 1.4, 95% CI = 0.9-2.0). Prostate cancer was also related positively to food and tobacco (OR= 2.1, 95% CI = 1.1-4.1), and chemical products (OR = 2.2, 95% CI = 0.7-7.2) industries. The analyses carried out to estimate the association between different types of pesticides and prostate cancer showed increased risks among farmers exposed to organochlorine insecticides and acaricides (OR = 2.5, 95% CI = 1.4-4.2), more specifically to the often contemporary used compounds DDT (OR = 2.1, 95% CI = 1.2-3.8), and dicofol (OR = 2.8, 95% CI = 1.5-5.0), whose effects could not be well separated. PMID- 12584744 TI - In vivo mapping of fractional plasma volume (fpv) and endothelial transfer coefficient (Kps) in solid tumors using a macromolecular contrast agent: correlation with histology and ultrastructure. AB - Contrast-enhanced MRI, immunostaining and electron microscopy were used to detect areas of intense angiogenesis in experimental tumors. This work was also aimed at evaluating the possible effect of the surrounding tissues on tumor microvasculature and at studying the penetration of macromolecules in avascular areas. Human colon carcinoma cells were implanted in subcutaneous tissue of nude mice. Dynamic T(1)-weigthed 3D pulse sequences were acquired before and after administration of Gd-DTPA-albumin to obtain parametric maps of fractional plasma volume (fpv) and transendothelial permeability (Kps). The maps suggested that tumor can be subdivided into 4 zones located in the peripheral rim (zones I-II) or in the core (zones III-IV) of the tumor itself. Significant differences (p<0.001) were found in the values of Kps and fpv of zones I-II with respect to zones III-IV. In the peripheral rim, permeability was significantly higher (p<0.01) in the muscle-peripheral region (zone I) with respect to the skin peripheral region (zone II). In areas with high Kps, histological and ultrastructural examination revealed clusters of newly formed vessels and signs of intense permeability. Numerous vascular vesicular organs were visible in these areas. In the tumoral core, analysis of the microcirculatory parameters revealed regions with mild permeability (zone III) and regions with negligible permeability (zone IV). These 2 zones were discriminated by the average value of Kps (p<0.05), while their fpv was not significantly different. Upon histological examination, the tumoral core exhibited necrotic areas; CD31 immunocytochemistry exhibited that it was diffusely hypovascularized with large avascular areas. Upon ultrastructural examination, capillaries were rarely visible and exhibited signs of endothelial cell damage. The results suggest that segmentation based on microvascular parameters detects in vivo zones characterized by immunocytochemical and ultrastructural aspects of intense angiogenesis. The finding that a certain amount of contrast agent penetrates in the tumoral core suggests that high oncotic and hydrostatic pressure only partially hinders the penetration of macromolecules. PMID- 12584745 TI - Usage of the NF-kappaB inhibitor sulfasalazine as sensitizing agent in combined chemotherapy of pancreatic cancer. AB - Sulfasalazine is commonly used as an anti inflammatory agent and is known as a potent inhibitor of NF-kappaB. Some pancreatic carcinomas are characterized by a constitutively elevated NF-kappaB activity accounting for chemoresistance. To elucidate whether blockade of NF-kappaB activity with sulfasalazine is suitable for overcoming this chemoresistance in vivo, we employed a mouse model with subcutaneously xenotransplanted human Capan-1 pancreatic carcinoma cells. Fourteen days upon tumor inoculation, animals were randomized in 6 groups, receiving no treatment, treatment with gemcitabine or with etoposide, either alone or in combination with sulfasalazine, or with sulfasalazine alone. Two therapy regimens were given with a 7-day interval in between. Upon treatment with etoposide or gemcitabine alone, tumor sizes were moderately reduced to 65-68% and 50-65%, respectively, as compared to untreated tumors. Sulfasalazine alone only decreased temporarily the tumor sizes. Sulfasalazine in combination with gemcitabine showed only partially higher reduction in tumor sizes than gemcitabine alone, whereas the combination with etoposide reduced significantly the tumor sizes in all experiments (down to 20%). TUNEL-staining showed higher numbers of apoptotic cells in tumors from the combination groups, in particular with etoposide, and proliferation as indicated by Ki67 staining was strongly reduced. Furthermore, combined treatment of sulfasalazine with the cytostatic drugs led to a decreased blood vessel density. Immunohistochemical staining of the activated p65 subunit showed that sulfasalazine treatment abolished the basal NF-kappaB activity in tumor xenografts. These data imply that the well established anti-inflammatory drug sulfasalazine sensitizes pancreatic carcinoma cells to anti cancer drugs, in particular to etoposide in vivo by inhibition of NF-kappaB. This combined chemotherapy offers great potential for improved anti tumor responses in pancreatic carcinomas. PMID- 12584746 TI - Optimal excitation-emission wavelengths for autofluorescence diagnosis of bladder tumors. AB - Tissue autofluorescence depends on endogenous fluorophores in the tissue, which undergo a change associated with malignant transformation. This change can be detected as an alteration in the spectral profile and intensity of autofluorescence. Our purpose was to determine the optimal excitation and emission wavelengths for autofluorescence diagnosis of bladder cancer. A total of 52 bladder tissue specimens were obtained from 25 patients undergoing mucosal biopsies or surgical resections of bladder tumors. Light-induced autofluorescence measurements were performed to study the spectroscopic differences between normal and malignant bladder tissue. Fluorescence excitation wavelengths varying from 220 to 500 nm were used to induce tissue autofluorescence, and emission spectra were measured in the 280-700 nm range. These spectra were then combined to construct 2-dimensional fluorescence excitation-emission matrices (EEMs). Significant changes in fluorescence intensity of EEMs were observed between normal and tumor bladder tissues, the most marked differences being at the excitation wavelengths of 280 and 330 nm. The diagnostic algorithm based on the combination of the fluorescence peak intensity ratios of I(350)/I(470) at 280 nm excitation and I(390)/I(470) at 330 nm excitation yielded a sensitivity of 100% [95% confidence interval (CI) 0.95-1.0] and specificity of 100% (95% CI 0.90 1.0). The results of the present fluorescence EEM study demonstrate that autofluorescence spectroscopy can distinguish malignant from normal bladder tissue and that excitation wavelengths of 280 and 330 nm are the most significant for differentiation between normal and malignant bladder mucosae with a high degree of diagnostic accuracy. PMID- 12584747 TI - Tumor-specific antigens in cutaneous T-cell lymphoma: expression and sero reactivity. AB - Cutaneous T-cell lymphoma (CTCL) is a heterogeneous group of extra-nodal non Hodgkin lymphomas with primary manifestation in the skin with poor treatment options in the advanced stages. As basis for future immune-therapeutic strategies we have investigated the possible expression of tumor-specific targets in CTCL focusing mainly on so-called cancer-germline genes. cDNAs derived from 20 CTCL tissues and 4 CTCL cell lines were tested with 15 gene-specific and 4 gene family specific primers by RT-PCR and confirmative Northern blotting. The most frequently detected mRNAs were LAGE-1 (55% with only partial coexpression of the splicing variants), cTAGE-1 (35%), MAGE-A9 (27%) and the GAGE-3-7 group (35%). Furthermore, we could detect NY-ESO-1 (21%) and a MAGE-A subgroup (15%), whereas sub-specification of the latter proved absence of MAGE-A1, -A2, -A3, -A6 and A12. SCP-1 was found in only one specimen and a several antigens could not been detected in any tumor tissue or cell line (MAGE-B, GAGE-1,2,8 and all 4 RAGE genes). 90% of all CTCL samples were positive for at least 1 of the frequent mRNAs in RT-PCR (LAGE-1, NY-ESO-1, cTAGE-1, MAGE-A9, or GAGE-3to7). Using a secondary SEREX approach we could detect sero-reactivity in sera of CTCL patients against recombinant cTAGE-1 (10/29), GAGE (3/19), MAGE-A1 (1/18), -A3 (1/18), -A6 (2/18) and -A9 (4/18) protein, but not against LAGE-1a, MAGE-A4b or MAGE-A12 protein (n = 19). We conclude, that certain cancer-germline genes can be detected frequently in CTCL and are able to elicit a systemic immune response. These candidate genes might therefore be promising targets for immunotherapeutic interventions in CTCL. PMID- 12584748 TI - Health profiles in 5836 long-term cancer survivors. AB - Increasingly, prolonged survival follows the diagnosis of cancer. Cancer therapies result in complex and lasting health effects that create unique health care needs for the survivors but are poorly understood (especially in survivors of adult cancers). Cancer survivors were asked to respond to a mailed health survey and provide medical and social information pertaining to their cancer experience. Information about demographics and perceived disease-related medical problems was analyzed. We analyzed the response of 5,836 survivors of adult cancers. Two-thirds of the responses came from women, and the response rate was 51% in both sexes. The mean interval since cancer diagnosis was 18.0 +/- 8.5 years. Younger survivors and men were more likely to report that cancer had affected their health. The health effect most commonly reported by survivors was arthritis/osteoporosis (26% of respondents). Survivors of Hodgkin's disease prominently reported thyroid and lung problems (33.8% of responders with the diagnosis). Prior diagnosis of lymphoma was associated with frequent mention of memory loss (14.7%). The passage of time decreased some perceived effects (memory loss) but increased others (arthritis/osteoporosis, cataracts). Compared with the general population, the incidence of several age- and gender-adjusted health conditions in cancer survivors is different. This group of cancer survivors reported generally good health but outlined multiple lasting medical problems. The health survey described represents 1 approach to the development of comprehensive information about the health needs of cancer survivors. PMID- 12584749 TI - Inhibition of integrin alpha5beta1 function with a small peptide (ATN-161) plus continuous 5-FU infusion reduces colorectal liver metastases and improves survival in mice. AB - Integrin alpha(5)beta(1) is expressed on activated endothelial cells and plays a critical role in tumor angiogenesis. We hypothesized that a novel integrin alpha(5)beta(1) antagonist, ATN-161, would inhibit angiogenesis and growth of liver metastases in a murine model. We further hypothesized that combining ATN 161 with 5-fluorouracil (5-FU) chemotherapy would enhance the antineoplastic effect. Murine colon cancer cells (CT26) were injected into spleens of BALB/c mice to produce liver metastases. Four days thereafter, mice were given either ATN-161 (100 mg/kg, every 3rd day) or saline by intraperitoneal injection, with or without combination of continuous-infusion 5-FU (100 mg/kg/2 weeks), which was started on day 7. On day 20 after tumor cell inoculation, mice were killed and liver weights and number of liver metastases were determined. A follow-up study on survival was also conducted in which mice were randomized to receive ATN-161, 5-FU or ATN-161+5-FU. Combination therapy with ATN-161+5-FU significantly reduced tumor burden (liver weight) and number of liver metastases (p<0.02). Liver tumors in the ATN-161 and ATN-161+5-FU groups had significantly fewer microvessels (p<0.05) than tumors in the control or 5-FU-treated groups. Unlike treatment with either agent alone, ATN-161+5-FU significantly increased tumor cell apoptosis and decreased tumor cell proliferation (p<0.03) and improved overall survival (p<0.03, log-rank test). Targeting integrin alpha(5)beta(1) in combination with 5 FU infusion reduced liver metastases formation and improved survival in this colon cancer model. The enhancement of antineoplastic activity from the combination of anti-angiogenic therapy and chemotherapy may be a promising approach for treating metastatic colorectal cancer. PMID- 12584750 TI - Disulfiram-mediated inhibition of NF-kappaB activity enhances cytotoxicity of 5 fluorouracil in human colorectal cancer cell lines. AB - 5-Fluorouracil (5-FU) is the major chemotherapeutic component for colorectal cancer (CRC) and other types of solid tumours. Resistance of cancer cells to 5-FU is considered the major obstacle for successful chemotherapy. NF-kappaB is a transcription factor. Cancer cells with high NF-kappaB nuclear activity demonstrate robust chemo- and radio-resistance. We demonstrated that nuclear NF kappaB activity in CRC cell lines, DLD-1 and RKO(WT), was significantly induced by 5-FU in a concentration- and time-dependent manner. 5-FU induced IkappaBalpha degradation and promoted both NF-kappaB nuclear translocation and its DNA binding activity. 5-FU treatment did not influence the activities of AP-1, AP-2, Oct-1, SP-1, CRE-B and TFIID. Disulfiram (DS), a clinically used anti-alcoholism drug, strongly inhibited constitutive and 5-FU-induced NF-kappaB activity in a dose dependent manner. DS inhibited both NF-kappaB nuclear translocation and DNA binding activity but had no effect on 5-FU-induced IkappaBalpha degradation. Used in combination, DS significantly enhanced the apoptotic effect of 5-FU on DLD-1 and RKO(WT) cell lines and synergistically potentiated the cytotoxicity of 5-FU to both cell lines. DS also effectively abolished 5-FU chemoresistance in a 5-FU resistant cell line H630(5-FU) in vitro. As DS has extensive preclinical and clinical experience, translating its anticancer usage from in vitro study to clinical trials is relatively straightforward. PMID- 12584751 TI - The molecular pharmacology of symplostatin 1: a new antimitotic dolastatin 10 analog. AB - Symplostatin 1, an analog of dolastatin 10, was recently isolated from cyanobacteria of the genus Symploca. Symplostatin 1 is a potent inhibitor of cell proliferation with IC(50) values in the low nanomolar range and it exhibits efficacy against a variety of cancer cell types. Symplostatin 1 caused the formation of abnormal mitotic spindles and accumulation of cells in metaphase at concentrations that had only minor effects on interphase microtubules. At higher concentrations, symplostatin 1 caused the loss of interphase microtubules. Cell cycle analysis revealed that symplostatin 1 caused G(2)/M arrest, consistent with its effects on mitotic spindles. Symplostatin 1 initiated the phosphorylation of Bcl-2, formation of micronuclei and activation of caspase 3, indicating induction of apoptosis. The cellular effects of symplostatin 1 are consistent with other antimitotic tubulin-targeting drugs. Tubulin polymerization experiments indicated that symplostatin 1 potently inhibits the assembly of purified tubulin, suggesting that tubulin may be its intracellular target. Some microtubule targeting agents are reported to have antiangiogenic activity and therefore the effects of symplostatin 1 on endothelial cell proliferation and invasion were evaluated. Symplostatin 1 was found to be a potent inhibitor of both endothelial cell proliferation and invasion. Because of its potent and broad activity in vitro, symplostatin 1 was evaluated in vivo. Symplostatin 1 was active against murine colon 38 and murine mammary 16/C; however, it was poorly tolerated and the mice were slow to recover from the toxicity. The data indicate that symplostatin 1 has a mechanism of action similar to dolastatin 10. PMID- 12584752 TI - Lack of serological evidence for an association between simian virus 40 and lymphoma. AB - Recent studies have implicated simian virus 40 (SV40) in non-Hodgkin's lymphomas based on detection of SV40 DNA sequences. We employed a virus-like-particle (VLP) based enzyme immunoassay for antibodies to SV40 to test sera from 520 lymphoma cases and 587 controls in Spain. The SV40 seroprevalence was 9.5% in controls and 5.9% in cases. Antibody levels of the positive sera were low. There was no association of SV40 seropositivity with any subtype of lymphoma. VLPs of the human BK virus substantially inhibited the SV40 reactivity of human sera. There was no serological evidence of widespread SV40 infection and no association of SV40 seropositivity with human lymphomas in Spain. PMID- 12584754 TI - Flow cytometric detection of beta-D-glucuronidase gene in wild-type bacterial cells using in-situ PCR. AB - An in situ PCR-based flow cytometry method useful for monitoring the presence or absence of the beta-D-glucuronidase gene in Escherichia coli has been developed. A single-step fixation and permeabilization procedure, which maintained cell integrity at the elevated temperatures used during thermal cycling in the presence of PCR reagents, was demonstrated. We have chosen a shorter DNA sequence of length 147 bp for the PCR. Cells subjected to in situ PCR using fluorescein-12 dUTP as a label, showed the presence of uid both in epifluorescence microscopic examination and flow cytometric analysis. Multi-parametric analysis of flow cytometric profiles revealed that the efficiency of labeling was found to be high. The potential of in situ PCR for the detection of uid in intact coliform cells was then successfully tested with a fecal coliform isolated from the coastal waters of Singapore. PMID- 12584755 TI - Effect of yeast extract on speciation and bioavailability of nickel and cobalt in anaerobic bioreactors. AB - The speciation of metals plays an important role in their bioavailability. In the case of anaerobic reactors for the treatment of wastewaters, the ubiquitous presence of sulfide leads to extensive precipitation of metals like nickel and cobalt, which are essential for the metabolism of the anaerobic microorganisms that carry out the mineralization of the pollutants present in the wastewater. In practice, nickel, cobalt, and iron are added in excessive amounts to full-scale installations. This study is concerned with the complexation of nickel and cobalt with yeast extract and its effect on the biogas production by methanogenic biomass. Adsorptive stripping voltammetry (AdSV) was used to get information about the stability and complexing capacity of the metal-yeast extract complexes formed. Nickel and cobalt form relatively strong organic complexes with yeast extract. The bioavailability of these essential metals in anaerobic batch reactors was dramatically increased by the addition of yeast extract. This is due to the formation of dissolved bioavailable complexes, which favors the dissolution of metals from their sulfides. Trace doses of yeast extract may be effective in keeping additions of essential metals to anaerobic reactors at a minimum. PMID- 12584756 TI - Free intracellular amino acid pools during autonomous oscillations in Saccharomyces cerevisiae. AB - In the present work dynamic changes of free intracellular amino acid pools during autonomous oscillations of Saccharomyces cerevisiae were quantified in glucose limited continuous cultivations. At a dilution rate of D = 0.22 h(-1) cyclic changes with a period of 120 min were found for many variables such as carbon dioxide production rate, dissolved oxygen, pH, biomass content, and various metabolite concentrations. On the basis of the observed dynamic patterns, free intracellular amino acids were classified to show oscillatory, stationary, or chaotic behavior. Amino acid pools such as serine, alanine, valine, leucine, or lysine were subjected to clear oscillations with a frequency of 120 min, identical to that of other described cultivation variables, indicating that there is a direct correlation between the periodic changes of amino acid concentrations and the metabolic oscillations on the cellular level. The oscillations of these amino acids were unequally phase-delayed and had different amplitudes of oscillation. Accordingly, they exhibited different patterns in phase plane plots vs. intracellular trehalose. Despite the complex and marked metabolic changes during oscillation, selected intracellular amino acids such as histidine, threonine, isoleucine, or arginine remained about constant. Concentrations of glutamate and glutamine showed a chaotic behavior. However, the ratio of glutamate to glutamine concentration was found to be oscillatory, with a period of 60 min and a corresponding figure eight-shaped pattern in a plot vs. trehalose concentration. Considering the described diversity, it can be concluded that the observed periodic changes are neither just the consequence of low or high rates of protein biosynthesis/degradation nor correlated to changing cell volumes during oscillation. The ratio between doubling time (189 min) and period of oscillation of intracellular amino acids (120 min) was 1:6. The fact that there is a close relationship between doubling time and period of oscillation underlines that the described autonomous oscillations are cell-cycle-associated. PMID- 12584757 TI - Metabolic analysis of the synthesis of high levels of intracellular human SOD in Saccharomyces cerevisiae rhSOD 2060 411 SGA122. AB - The synthesis of human superoxide dismutase (SOD) in batch cultures of a Saccharomyces cerevisiae strain using a glucose-limited minimal medium was studied through metabolic flux analysis. A stoichiometric model was built, which included 78 reactions, according to metabolic pathways operative in these strains during respirofermentative and oxidative metabolism. It allowed calculation of the distribution of metabolic fluxes during diauxic growth on glucose and ethanol. Fermentation profiles and metabolic fluxes were analyzed at different phases of diauxic growth for the recombinant strain (P+) and for its wild type (P ). The synthesis of SOD by the strain P+ resulted in a decrease in specific growth rate of 34 and 54% (growth on glucose and ethanol respectively) in comparison to the wild type. Both strains exhibited similar flux of glucose consumption and ethanol synthesis but important differences in carbon distribution with biomass/substrate yields and ATP production 50% higher in P-. A higher contribution of fermentative metabolism, with 64% of the energy produced at the phosphorylation level, was observed during SOD production. The flux of precursors to amino acids and nucleotides was higher in the recombinant strain, in agreement with the higher total RNA and protein levels. Lower specific growth rates in strain P+ appear to be related to the decrease in the rate of synthesis of nonrecombinant protein, as well as a decrease in the activities of the pentose phosphate (PP) pathway and TCA cycle. A very different way of entry into the stationary phase was observed for each strain: in the wild-type strain most metabolic fluxes decreased and fluxes related to energy reserve synthesis increased, while in the P+ strain the flux of 22 reactions (including PP pathway and amino acids biosynthesis) related to SOD production increased their fluxes. Changes in SOD production rates at different physiological states appear to be related to the differences in building blocks availability between respirofermentative and oxidative metabolism. Using the present expression system, ideal conditions for SOD synthesis are represented by either active growth during respirofermentative metabolism or transition from a growing to a nongrowing state. An increase in SOD flux could be achieved using an expression system nonassociated to growth and potentially eliminating part of the metabolic burden. PMID- 12584758 TI - Microalgae cultivation in air-lift reactors: modeling biomass yield and growth rate as a function of mixing frequency. AB - The slow development of microalgal biotechnology stems from the failure in the design of large-scale photobioreactors where light energy is efficiently utilized. Due to the light gradient inside the reactor and depending on the mixing properties, algae are subjected to certain light/dark cycles where the light period is characterized by a light gradient. These light/dark cycles will determine productivity and biomass yield on light energy. Air-lift reactors can be used for microalgae cultivation and medium-frequency light/dark cycles will be found in these systems. Light/dark cycles are associated with two basic parameters: first, the light fraction, i.e., the ratio between the light period and the cycle time and second, the frequency of the light/dark cycle. In the present work, light/dark cycles found in air-lift reactors were simulated taking into account the light gradient during the light period. The effect of medium frequency cycle time (10-100 s) and light fraction (0.1-1) on growth rate and biomass yield on light energy of the microalgae Dunaliella tertiolecta was studied. The biomass yield and growth rates were mainly affected by the light fraction, while cycle time had little influence. Response surface methodology was used and a statistical model describing the effect of light fraction and cycle time on growth rate and biomass yield on light energy was developed. The use of the model as a reactor design criterion is discussed. PMID- 12584759 TI - A light distribution model for an internally radiating photobioreactor. AB - Analysis of light energy distribution in culture is important for maximizing the growth efficiency of photosynthetic cells and the productivity of a photobioreactor. To characterize the irradiance conditions in a photobioreactor, we developed a light distribution model for a single-radiator system and then extended the model to multiple radiators using the concept of parallel translation. Mathematical expressions for the local light intensity and the average light intensity were derived for a cylindrical photobioreactor with multiple internal radiators. The proposed model was used to predict the irradiance levels inside an internally radiating photobioreactor using Synechococcus sp. PCC 6301 as a model photosynthetic microorganism. The effects of cell density and radiator number were interpreted through photographic and model simulation studies. The predicted light intensity values were found to be very close to those obtained experimentally, which suggests that the proposed model is capable of accurately interpreting the local light energy profiles inside the photobioreactor system. Due to the simplicity and flexibility of the proposed model, it was also possible to predict the light conditions in other complex photobioreactors, including optical-fiber and pond-type photobioreactors. PMID- 12584760 TI - Immobilization of thermolysin to polyamide nonwoven materials. AB - In the last few years, an increasing number of biotechnological techniques have been applied to the restoration and conservation of works of art, paintings, old maps, and papers or books. Enzymes can solve problems that give restorers difficulties, although for many applications it is not possible to use soluble enzymes; therefore, it is necessary to look for suitable carriers for immobilization. Different methods for covalent immobilization of enzymes to polyamide nonwovens were tested, using thermolysin as an example. Two distinct strategies were pursued: (1). controlled, partial hydrolysis of the polymer and subsequent binding of the enzyme to the released amino and carboxy groups; and (2). attachment of reactive groups directly to the polyamide without disintegrating the polymeric structure (O-alkylation). Different spacers were used for covalent fixation of the enzyme in both cases. The enzyme was fixed to the released amino groups by glutaraldehyde, either with or without a spacer. Either way, active enzyme could be immobilized to the matrix. However, intense treatment caused severe damage to the stability of the nonwoven fabric, and reduced the mechanical strength. Conditions were investigated to conserve the nonwoven fabric structure while obtaining near-maximum immobilized enzyme activity. Immobilization of the enzyme to the released carboxy group after acid hydrolysis was performed using dicyclohexylcarbodiimide. In comparison to the enzyme bound via the amino group, the yield of immobilized enzyme activity was slightly lower when benzidine was taken as spacer and still lower with a 1,6 hexanediamine spacer. O-alkylation performed with dimethylsulfate caused severe damage to the nonwoven fabric structure. Considerably better results were obtained with triethyloxonium tetrafluoroborate. As the spacers 1,6-hexanediamine and adipic acid dihydrazide were used, activation for immobilizing thermolysin was performed with glutaraldehyde, adipimidate, and azide. With the exception of azide, all combinations of spacers and activation reagents gave high yields of immobilized enzyme activity. Thermolysin immobilized by this technique showed a remarkably improved stability with respect to elevated temperature, extreme pH values, and reduced polarity. The nonwoven fabric can be stored for weeks without loss of enzyme activity by washing with distilled water and drying. PMID- 12584762 TI - Compression of biocompatible liquid-filled HSA-alginate capsules: determination of the membrane mechanical properties. AB - Compression experiments between two parallel plates are performed on a series of biocompatible HSA-alginate capsules with two different membrane thicknesses. The capsule geometry and size as well as the average membrane thickness are first measured. The compression set-up is fitted with a sensitive force transducer that allows measurement of the compression force as a function of plate separation. The response of the capsule is analyzed by assuming different constitutive models for the membrane, where the shear and surface dilatation effects are accounted. An apparent area dilatation modulus is then computed for different values of the plate separation and required to remain constant as the capsule deformation increases. This allows identification of plausible constitutive laws for the membrane material. PMID- 12584761 TI - Size-exclusion reaction chromatography (SERC): a new technique for protein PEGylation. AB - A new, widely applicable process that combines reaction and separation in a single unit operation is described. The process, size-exclusion reaction chromatography (SERC), simultaneously allows control of the extent of reactions in which molecular size is altered and the separation of products and reactants. In SERC, a moving reaction zone is formed by injection of reactants onto a size exclusion chromatography column. Reactants and products are partitioned differently within the mobile phase, resulting in different linear flow rates through the column. The products are therefore removed selectively from the reaction zone, minimizing their residence time in the reaction zone and allowing their separation in the downstream section of the column. For reactions such as protein PEGylation, in which successive addition of PEG groups to the protein results in significant molecular size increases, SERC potentially offers a method by which a dominant final PEGylated protein size can be produced at high yield. The SERC PEGylation of two model proteins, alpha-lactalbumin and beta lactoglobulin, is demonstrated and results show that simultaneous reaction and separation was obtained. PMID- 12584763 TI - High cell density cultivation of probiotics and lactic acid production. AB - The commercial interest in functional foods that contain live microorganisms, also named probiotics, is paralleled by the increasing scientific attention to their functionality in the digestive tract. This is especially true of yogurts that contain strains of lactic-acid bacteria of intestinal origin, among these, Lactobacillus delbrueckii ssp. bulgaricus is extensively used in the dairy industry and it has been demonstrated to be a probiotic strain. In this work we describe high cell density cultivations of this microorganism also focusing on the stereospecific production of lactic acid. Key parameters such as medium composition (bactocasitone concentration) and diverse aeration conditions were explored. The results showed that the final concentration of biomass in anaerobic fermentation was lower than the one obtained in microaerophilic conditions, while it gave a very high productivity of lactic acid which was present as a racemic mixture in the permeate. Fermentation experiments carried out with air sparging, even at very low flow-rate, led to the production of the sole L(+) lactic acid giving sevenfold increase in biomass yield in respect to the batch cultivation. Finally, a mathematical model was developed to describe the microfiltration bioprocess applied in this research considering an inhibition kinetic and enucleating a suitable mathematical description for the decrease of the transmembrane flux. PMID- 12584764 TI - Oxidation of 4-bromophenol by the recombinant fused protein cellulose-binding domain-horseradish peroxidase immobilized on cellulose. AB - A fused protein consisting of cellulose-binding domain (CBD) and horseradish peroxidase (HRP) was constructed and expressed in Escherichia coli. Refolded recombinant CBD-HRP (95% recovery yield) was bound to microcrystalline cellulose and applied for the oxidation of a model toxic phenol, 4-bromophenol (BP). Oxidation of BP by CBD-HRP resulted in the formation of dimers to pentamers as evidenced by mass spectrometry analysis. When immobilized, the vast majority of the oxidation products adsorbed to the cellulose matrix. CBD-HRP (0.75 pyrogallol units) bound to 0.1 g cellulose was packed in a column, connected to an HPLC pump and monitoring system, and column performance and capacity were studied under various operating conditions. When performance was studied as a function of BP loading rate at a constant H(2)O(2) loading rate of 1500 nmol/min, V(app) (max) and K(m) (app) were calculated to be 5.29 +/- 0.46 micromol mL min and 644.9 +/- 114.3 microM, respectively. Immobilized CBD-HRP exhibited enhanced stability to H(2)O(2) and oxidized considerably more BP than free CBD-HRP. Inclusion of gelatin, which suppresses product-dependent inactivation, further increased the amount of BP oxidation. These findings may have potential impact in terms of enzyme supply in high-rate treatment of wastewater contaminated with toxic phenols, since the susceptibility of peroxidases to both H(2)O(2) - and product dependent inactivation demands continuous supply of fresh enzyme. PMID- 12584765 TI - Self-assembly of Pseudomonas fluorescens lipase into bimolecular aggregates dramatically affects functional properties. AB - It has been found that lipase from Pseudomonas fluorescens (PFL) is able to aggregate into bimolecular structures (MW around 66 kD) even at moderate enzyme concentrations. At very low enzyme concentrations and in the presence of detergents, the same enzyme displayed a unimolecular structure with a molecular weight of 33 kD. Both enzyme structures displayed different functional properties. First, the bimolecular structure was much more stable than the unimolecular species (the bimolecular structure maintained over 80% of initial activity after 72 hours at 45 degrees C, while the unimolecular structure retained only around 30% of initial activity after 4 hours of incubation under the same experimental conditions); and the bimolecular form presented a higher optimal T. Second, the unimolecular form showed a much lower K(M) for ethyl butyrate than the bimolecular form. Third, the interfacial activation in biphasic substrate-aqueous milieu was higher for the bimolecular form. Fourth, the unimolecular structure was less active but much more enantioselective than the unimolecular species in the model reaction used. It is proposed that the bimolecular aggregates of PFL might be formed by two open lipase molecules (mutual interfacial activation), in intimate contact, and that the bimolecular form represents an example of "pseudo-quaternary" structure. PMID- 12584767 TI - Working in a climate of innovation. PMID- 12584766 TI - The effect of dissolved oxygen on PHB accumulation in activated sludge cultures. AB - Nitrogen removal from wastewater is often limited by the availability of reducing power to perform denitrification, especially when treating wastewaters with a low carbon:nitrogen ratio. In the increasingly popular sequencing batch reactor (SBR), bacteria have the opportunity to preserve reducing power from incoming chemical oxygen demand (COD) as poly-beta-hydroxybutyrate (PHB). The current study uses laboratory experiments and mathematical modeling in an attempt to generate a better understanding of the effect of oxygen on microbial conversion of COD into PHB. Results from a laboratory SBR with acetate as the organic carbon source showed that the aerobic acetate uptake process was oxygen-dependent, producing higher uptake rates at higher dissolved oxygen (DO) supply rates. However, at the lower DO supply rates (k(L)a 6 to 16 h(-1), 0 mg L(-1) DO), a higher proportion of the substrate was preserved as PHB than at higher DO supply rates (k(L)a 30, 51 h(-1), DO >0.9 mg L(-1)). Up to 77% of the reducing equivalents available from acetate were converted to PHB under oxygen limitation (Y(PHB/Ac) 0.68 Cmol/Cmol), as opposed to only 54% under oxygen-excess conditions (Y(PHB/Ac) 0.48 Cmol/Cmol), where a higher fraction of acetate was used for biomass growth. It was calculated that, by oxygen management during the feast phase, the amount of PHB preserved (1.4 Cmmol L(-1) PHB) accounted for an additional denitrification potential of up to 18 mg L(-1) nitrate-nitrogen. The trends of the effect of oxygen (and hence ATP availability) on PHB accumulation could be reproduced by the simulation model, which was based on biochemical stoichiometry and maximum rates obtained from experiments. Simulated data showed that, at low DO concentrations, the limited availability of adenosine triphosphate (ATP) prevented significant biomass growth and most ATP was used for acetate transport into the cell. In contrast, high DO supply rates provided surplus ATP and hence higher growth rates, resulting in decreased PHB yields. The results suggest that oxygen management is crucial to conserving reducing power during the feast phase of SBR operation, as excessive aeration rates decrease the PHB yield and allow higher biomass growth. PMID- 12584768 TI - Tobacco or health? PMID- 12584769 TI - Reaffirming the need for infection control. PMID- 12584770 TI - Characteristics of and current treatment options for hepatitis B and hepatitis C. PMID- 12584771 TI - Waterborne biofilms and dentistry: the changing face of infection control. AB - Interest in and concern about the biofilms that occur in dental equipment and waterlines have been increasing in recent years. Dental unit waterlines are ideal environments for the growth of microorganisms entering dental units from the municipal water supply. This article describes the conditions in waterline tubing that favour development of biofilms and discusses the level of risk that such microbial growth poses for both dental professionals and their patients. It is stressed that very few cases of infection have been linked directly to contamination in dental unit waterlines. Finally, potential solutions for minimizing risks are presented and discussed. PMID- 12584772 TI - Risk management--practical considerations. PMID- 12584773 TI - Management of exposure to bloodborne pathogens. PMID- 12584774 TI - Hand hygiene: washing and disinfection. PMID- 12584775 TI - Risk of bacterial transmission in dental practice. PMID- 12584776 TI - Hepatitis B immunization and postimmunization serology. PMID- 12584777 TI - Risk of transmission of viruses in the dental office. PMID- 12584778 TI - Universal precautions. PMID- 12584779 TI - Disinfection of surfaces and equipment. PMID- 12584781 TI - [Retrospection on the 25 years of persisting integrated medicine by studying of rhubarb as a breakthrough]. PMID- 12584780 TI - Unconventional dentistry: Part IV. Unconventional dental practices and products. AB - This is the fourth in a series of 5 articles providing a contemporary overview and introduction to unconventional dentistry (UD) and its correlation to unconventional medicine (UM). Several common UD and UM practices are described to familiarize practitioners with a variety of theories, practices, products and treatments that specifically apply to dentistry. This brief review is not intended as an in-depth resource. PMID- 12584782 TI - [Consideration for the study of basic theory of TCM]. PMID- 12584783 TI - [Assessment on effect of treatment for childhood hyperkinetic syndrome by combined therapy of yizhi mixture and ritalin]. AB - OBJECTIVE: To seek for a good therapy for childhood hyperkinetic syndrome (CHS). METHODS: Two hundred and ten patients of CHS were randomly divided into three groups, the Yizhi mixture (YZM) group, the Ritalin group and the combined treated group, 70 in each group. They were treated with YZM alone, Ritalin alone and combined YZM and Ritalin respectively for 12 weeks. The effect was assessed by Conners' questionnaire for hyperkinetic scoring, Achenbach's questionnaire for behavior estimation, clinical general image questionnaire and Tess's questionnaire before and after treatment. RESULTS: The total effective rate in the YZM group, Ritalin group and combined treated group was 75.7%, 74.2% and 82.8% respectively, and the markedly effective rate in the three groups was 44.3%, 45.7% and 80.0% respectively. By chi 2 test, comparison among the three groups showed that the difference of the total effective rate was insignificant, while that of the markedly effective rate was significant (chi 2 = 23.31, P < 0.01). Further analysis conducted by partitioning method to make comparison between each two groups showed that the markedly effective rate was significantly higher in the combined treated group than that in the YZM group (chi 2 = 18.98, P < 0.01) and in the Ritalin group (chi 2 = 17.62, P < 0.01). The low hyperkinetic score, behavior score, achievement of Chinese and mathematics in the CHS patients were improved after treatment, the improvements were significantly higher in the combined treated group than those in the other two groups (P < 0.05, by q-test). Moreover, the soft nerve signs and abnormal encephalogram were significantly improved in the combined treated and the YZM groups, and the side-effect occurred in the two groups was less than that in the Ritalin group (P < 0.01). CONCLUSION: The therapeutic effect of combined treatment of YZM and Ritalin was better than that of YZM or Ritalin alone. PMID- 12584785 TI - [Clinical observation on treatment of gastrointestinal dysfunction by fu'an liquid for retention enema in children with critical illness]. AB - OBJECTIVE: To observe the therapeutic effect of Fu'an Liquid (FAL) for retention enema in treating gastrointestinal (GI) dysfunction of children with critical illness. METHODS: Eighty-nine patients were randomly divided into two groups, 52 in the treated group and 37 in the control group. Conventional therapy of western medicine was given to both groups and to the treated group FAL was given additionally. Plasma endothelin (ET) level was measured during admission, GI dysfunction occurrence and after treatment, and the therapeutic effect as well as the recovery of GI condition were observed. RESULTS: The total effective rate of FAL in treating GI dysfunction was 84.62%, which was significantly higher than that in the control group (62.17%) (P < 0.05). In the treated group, 34 cases were treated successfully, 16 died and the other 2 abandoned, the mortality rate being 30.77%, while in the control group, the corresponding numbers were 16, 18, 3 and 48.65%. The mortality rate in the treated group was lower than that in the control group (chi 2 = 4.64, P < 0.05). Level of ET in both groups was higher than normal range during admission (P < 0.01), it further increased when GI dysfunction occurred (P < 0.01), and decreased when successfully treated, the decrease was quicker in the treated group than that in the control group (P < 0.05). CONCLUSION: In children with critical illness, ET level would increase when the patient was complicated with GI dysfunction. FAL for retention enema could reduce the ET level effectively, promote the recovery of patients from GI dysfunction, so as to play a definite role in enhancing the successful rate of rescue. PMID- 12584784 TI - [Clinical study on treatment of children attention deficit hyperactivity disorder by jiangqian granule]. AB - OBJECTIVE: To explore the effect of Jiangqian granule (JQG) in treating children attention deficit hyperactivity disorder (ADHD) and its mechanism. METHODS: Eighty children with ADHD were randomly divided into two groups, the treated group treated with JQG and the control group treated with Ritalin. Blood lead concentration and Conners' scoring were measured before and after 3 months treatment and the therapeutic effects were assessed. Besides, blood lead concentration and Conners' scoring in 60 healthy children were also determined for control. RESULTS: The total effective rate in the treated group and the control group was 92.5% and 72.5% respectively, significant difference existed between the two groups. Blood lead level and Conners' scores in ADHD patients were significantly higher than those in the healthy control (P < 0.01). The two parameters decreased after treatment in both groups, but the lowering was more significant in the treated group (P < 0.05, P < 0.01). CONCLUSION: Blood lead concentration raised in ADHD patients, JQG could enhance the clinical therapeutic effect by means of reducing the blood lead. PMID- 12584787 TI - [Effect of folium ginkgo extract on the erythrocyte immunity function and serum lipid peroxide in asphyxia neonate]. AB - OBJECTIVE: To observe the changes of erythrocyte immunity and serum lipid peroxide in asphyxia neonate, and to study the effect of Folium Ginkgo extract (FGE) on them. METHODS: Thirty asphyxia neonates were randomly divided into 2 groups, the treated group and the control group, 15 in each group. Erythrocyte C3b receptor rosette rate (E-C3bRR), erythrocyte immune complex rosette rate (E ICR), blood superoxide dismutase (SOD) activity and serum lipid peroxide (LPO) level were determined at 24 hrs after birth. Conventional treatment was given to both groups and FGE (15 mg/kg.d) was given to the treated group additionally for 7-8 days, then the above-mentioned parameters were re-examined and neonatal behavioral neurological assessment (NBNA) was measured as well. RESULTS: E-C3bRR and SOD lowered, E-ICR and serum LPO increased in the asphyxia neonate significantly (P < 0.05). After treatment, comparison between the two groups showed that E-C3bRR and SOD were higher, E-ICR and serum LPO were lower in the treated group than those in the control group, and NBNA scoring was obviously higher in the former than that in the latter (all P < 0.05). CONCLUSION: Decrease of erythrocyte immunity in asphyxia neonate is related to the declined anti oxidation ability and lipid peroxidase injury. FGE could suppress the free radical production, scavenge free radicals, antagonize the lipid peroxidation injury of cell membrane and up-regulate erythrocyte immunity. It displays the effects of nerve tissue protection and hypoxia-ischemic brain injury alleviation. PMID- 12584786 TI - [Effect of rhubarb on inflammatory cytokines and complements in patients with systemic inflammation reaction syndrome and its significance]. AB - OBJECTIVE: To study the effect of rhubarb in treating patients with systemic inflammation reaction syndrome (SIRS) and its mechanism. METHODS: The 40 patients with SIRS in the treated group were treated with conventional treatment plus rhubarb powder orally or by nasal feeding, the 38 patients in the control group were treated with conventional treatment alone. Serum tumor necrosis factor-alpha (TNF-alpha) was determined by ELISA during the admission and the 3rd day after admission, C-reactive protein (C-RP), complement 3 and 4 (C3, C4) were also determined by auto-scattering turbidimetric quantitative analysis. The parameters were compared between groups and with normal control group. RESULTS: Cure rate in the treated group was significantly higher than that in the control group accompanied with lesser occurrence of multiple organ dysfunction syndrome (MODS) and lower mortality. Serum TNF-alpha, C-RP, C3 and C4 in the SIRS patients were increased during admission, which were significantly higher than normal control, but these parameters would be reduced together with the alleviating of symptoms after treatment, particularly after rhubarb treatment. CONCLUSION: Rhubarb could improve the prognosis of patients with SIRS, its major mechanism is that rhubarb has the antagonizing effect against inflammatory cytokines and complements. PMID- 12584788 TI - [Study on relationship of clinical effect of xuefu zhuyu concentrated pill and serum pharmacological anti-platelet activating effect in vitro]. AB - OBJECTIVE: To assess the significance of platelet activation in unstable angina pectoris (UA) and acute myocardial infarction (AMI), and to explore the relationship of clinical effect of Xuefu Zhuyu concentrated pill (XCP) in vivo and the serum pharmacological anti-platelet activating effect of XCP in vitro. METHODS: By flow cytometry and special labelled antibodys to detect the whole blood platelet membranous glycoprotein CD62P and CD41/45 expressions in UA patients before and after XCP treatment, as well as those in peripheral blood of AMI rats before and after co-cultured with XCP containing serum from patients at 37 degrees C for 30 min. RESULTS: CD62P and CD41/45 expressions increased significantly in UA patients to 24.36 +/- 7.91% and 29.51 +/- 12.21% respectively. After effective treatment, they decreased to 19.57 +/- 7.22% and 20.87 +/- 8.73% respectively accompanied with increase of platelet adhesion and aggregation rate. The difference of CD62P before and after treatment was significant (P < 0.05). CD62P in blood of AMI rats was 39.73 +/- 12.36%, after being co-cultured with XCP containing serum from patients treated effectively, it reduced to 30.41 +/- 10.36% (P < 0.05), but after co-cultured with the serum from patients treated ineffectively, it showed less intervention effect (P > 0.05). CONCLUSION: Platelet was highly activated in UA patients and AMI rats. The consistency between clinical holistic effect of XCP and serum pharmacological effect of XCP in platelet activation inhibition reflects a good correlation between serum pharmacological effect of Chinese herbal medicine and its clinical holistic effect. PMID- 12584789 TI - [Study on effect of jiangya tongmai recipe on vascular activating substances in patients of hypertension with left ventricular hypertrophy]. AB - OBJECTIVE: To study the effect of Jiangya Tongmai Recipe (JYTMR), a Chinese herbal medicine preparation for activating blood circulation to remove stasis, on vascular activating substances in treating patients of hypertension with left ventricular hypertrophy. METHODS: The 37 patients with hypertension were randomly divided into two groups, the treated group (n = 21) and the control group (n = 16). They were treated with JYTMR and captopril respectively for 8 weeks. Left ventricular mass weight (LVMI), peak flow velocity of early diastole (Emax), peak flow velocity of atrial contraction (Amax), Emax/Amax, ejection fraction (EF), as well as levels of plasma endothelin (ET), calcitonin gene-related peptide (CGRP) and angiotensin II (Ang II) were measured before and after treatment and compared. RESULTS: JYTMR could enhance the left ventricular dilation and contractile functions, lower the levels of plasma ET and Ang II and increase the level of plasma CGRP. CONCLUSION: JYTMR shows good effect in improving left ventricular function and regulating vascular activating substances, it could prevent and treat hypertension and its complications for prolonged treatment via multiple paths and links. PMID- 12584790 TI - [Effect of shengmai injection on hemodynamics in patients with dilated cardiomyopathy]. AB - OBJECTIVE: To observe the effects of Shengmai Injection (SMI) on hemodynamics in patients with dilated cardiomyopathy (DCM), and to explore the clinical effect of SMI in treating patients of DCM with heart failure. METHODS: One hundred patients were divided into two groups. In the 50 cases of the treated group, 10 cases with heart function of II degree, 35 of III degree and 5 of IV degree. The corresponding number of cases in the 50 patients of the control group were 10, 36 and 4. Conventional treatment was given to both groups, and SMI was administered to the treated group additionally. The therapeutic effect and the indexes of heart function before and after treatment were determined and compared. RESULTS: In the treated group, treatment showed markedly effective in 22 cases and effective in 20, the total effective rate being 84%, while in the control group, markedly effective in 14 and effective in 16, the total effective rate being 60%, the comparison between the two groups showed significant difference (chi 2 = 7.14, P < 0.01). In the treated group, cardiac output, stroke volume (SV), cardiac index, eject fraction (EF), lefe ventricular minor axis shortened rate, ventricular wall thickened rate were all increased after treatment and system vascular resistance (SVR) decreased significantly (P < 0.05), while in the control group, insignificant change was found in the above-mentioned parameters after treatment (P > 0.05). Comparison between the two groups after treatment showed that EF and SV were obviously higher and SVR obviously lower in the treated group than those in the control group. CONCLUSION: SMI could markedly improve the heart function of patients with DCM. The effect of conventional treatment would be enhanced in combination therapy with SMI. PMID- 12584791 TI - [Study on relationship between Arg778Leu/Gln gene mutation spot in ATP7B and TCM syndrome type in Chinese patients with Wilson disease]. AB - OBJECTIVE: To investigate the relationship between Arg778Leu/Gln gene mutation spot in ATP7B and TCM Syndrome type in Chinese patients with Wilson disease (WD). METHODS: Exon 8 of ATP7B of 90 WD patients and 30 healthy controls were amplified by PCR and analysed by restriction enzyme Msp I, the TCM Syndrome type of the patients was differentiated at the same time. RESULTS: In the 90 WD patients, 34 with Arg778Leu/Gln of exon 8 were detected, among them 20 cases belonged to the TCM Syndrome type of endogenous Liver-Wind agitation. CONCLUSION: Onset age of WD patients with Arg778Leu/Gln mutation is later than that without this mutation. Arg778Leu/Gln mutation might be related to the TCM Syndrome type of endogenous Liver-Wind agitation. PMID- 12584793 TI - [Study of protective effect and mechanism for baicalin on bacillus pertussis infected brain tissue and its dose-effect relationship]. AB - OBJECTIVE: To study the possible mechanism of protective effect for Baicalin on Bacillus pertussis (BP) infected brain tissue and the dose-effect relationship. METHODS: Brain tissues slices were divided into 7 groups: (1) the normal group; (2) the model group: infected by 10% BP; (3) the baicalin group, which was pretreated with baicalin, infected by BP and subdivided into 5 sub-groups according to different doses of baicalin used; (4) the glutamic acid group: cultured with glutamic acid; (5) the baicalin plus glutamic acid group; (6) the peroxide group: cultured with hydrogen peroxide; and (7) the baicalin plus peroxide group. The lactate dehydrogenase (LDH) content in the supernatant of culture was determined and quantitative protein determination was conducted. RESULTS: The LDH releasing was higher in the model group, glutamic acid group and peroxide group as compared with that in the normal group, 15.10 +/- 4.89 u/g. protein (the same unit below), 15.49 +/- 5.66 and 16.54 +/- 5.47 vs 6.10 +/- 2.87 respectively (P < 0.01). After being pretreated with 0.25 mmol/L baicalin, LDH level decreased significantly to 8.65 +/- 2.43, which was significantly different from that in the model group (P < 0.01), LDH was also decreased in the baicalin plus glutamic acid group (9.93 +/- 2.89) and baicalin plus peroxide group (9.54 +/- 2.82), which was significantly lower than that in the glutamic acid group and the peroxide group respectively (P < 0.01). CONCLUSION: Pretreatment of baicalin has protective effect on BP caused nerve cell injury in rat brain slices, the protection is possibly related with the reduction of glutamic acid and hydrogen peroxide induced damage on nerve cells in vitro. PMID- 12584792 TI - [Experimental study on inhibitory effect of rutin against platelet activation induced by platelet activating factor in rabbits]. AB - OBJECTIVE: To explore the inhibitory effect and mechanism of rutin against platelet activating factor (PAF) induced platelet aggregation, 5-HT release and intra-platelet free calcium concentration. METHODS: The rate of washed rabbit platelet (WRP) aggregation was measured by turbidimetry and O-phthaldialdehyde (OPT) fluoro-spectrophotometry (FSPM) was used to determine 5-HT content. The intraplatelet free calcium concentration was measured with Fura-2/AM FSPM assay. RESULTS: Rutin in vitro was concentration-dependently inhibiting PAF (9.55 x 10( 9) mol/L) induced WRP aggregation, the IC50 of 5-HT release was 0.73, 1.13 mmol/L respectively and the intraplatelet free calcium concentration elevation evoked by PAF (4.78 x 10(-10) mol/L) were inhibited by 68.3, 136, 274, 545 mumol/L of rutin dose-dependently. CONCLUSION: Rutin could inhibit PAF induced platelet aggregation, 5-HT release and the increase of intraplatelet free calcium. PMID- 12584794 TI - [Effects of bazheng mixture on urinary amount and contractile-relaxant function of isolated urethral smooth muscle in rabbits]. AB - OBJECTIVE: To explore the pharmacological mechanism of Bazheng Mixture (BZM) in promoting diuresis and relieving stranguria. METHODS: By means of urine collection through indwelling catheter and urethral ring isolation test to observe the effects of BZM on the urinary amount of conscious rabbits and contractile-relaxant function of urethral smooth muscle isolated from rabbits. RESULTS: BZM, at dosage of 5.0 g/kg or 10.0 g/kg by gastrogavage, could increase the urinary amount in 60 min obviously (P < 0.05). For the isolated urethral rings, the maximal contractile and maximal relaxant forces reduced significantly (P < 0.05 or P < 0.01) when the concentration of BZM applied ranged between 9.9 90.9 g/L; the frequency increased significantly (P < 0.05 or P < 0.01) when the concentration ranged 29.1-90.9 g/L and the amplitude increased significantly when the concentration were 56.6 and 90.0 g/L. Along with the concentration of BZM increased from 9.9 g/L to 90.9 g/L, the changes occurred in the above-mentioned parameters of urethral ring were in order of potency amplitude > frequency > maximal relaxant force > maximal contractile force. CONCLUSION: The effects of BZM in enhancing urethral peristalsis may be stronger than that in dilating the urethral caliber. Its mechanism in promoting diuresis and relieving stranguria may be related with its action of urethral dilatation and potential peristalsis promotion. PMID- 12584795 TI - [Effect of arsenic trioxide on apoptosis of pulmonary eosinophile in asthmatic guinea-pigs]. AB - OBJECTIVE: To study the effect and mechanism of arsenic trioxide (As2O3) on apoptosis of pulmonary eosinophiles (PE) in asthmatic guinea-pigs. METHODS: Thirty guinea-pigs were divided into 3 groups at random, the control group, the asthmatic group and the As2O3 group. The dosage of As2O3 used was 2 mg/kg. The apoptotic PE were labelled by TdT-mediated dUTP nick end labelling technique, and the PE infiltration and apoptosis were detected quantitatively using computerized image analysis technique. RESULTS: In the control group, the amount of infiltrating PE was 4.4 +/- 2.5 cells/HP and the PE apoptotic index (AI) was 0.42 +/- 0.08%. In the asthmatic group, the amount increased (P < 0.01) and AI decreased significantly (P < 0.01). After the asthmatic animals had been treated with As2O3, the two parameters changed reversedly significantly (P < 0.01), and there was a significantly negative correlation between them (r = -0.949, P < 0.01). CONCLUSION: The PE apoptosis abnormality is one of the important mechanisms that cause bronchial asthma, As2O3 could alleviate the airway inflammation through promoting PE apoptosis and lower PE infiltration. Low dose of As2O3 is proved to be effective with relative safety, it also has potential value in treating asthma. PMID- 12584796 TI - [Effect of electro-acupuncture on calcium content in neurocytes of focal cerebral ischemia]. AB - OBJECTIVE: To study the effect of electro-acupuncture (EA) in regulating calcium (Ca2+) content in brain neurocytes of rats with focal cerebral ischemia. METHODS: The changes of Ca2+ content in ischemic neurocytes were observed by using laser confocal scanning microscope. RESULTS: Ca2+ content did not change significantly in cerebral cortex when the brain ischemia occurred for 1 hr, but it raised significantly in striatum neurocytes. Both the Ca2+ contents in striatum and cortex area increased significantly 3 hrs after ischemia occurrence and the content in striatum was higher than that in cortex significantly. Brain Ca2+ content could be reduced significantly after the 3 hrs ischemic brain were treated by EA for 30 min. CONCLUSION: EA could regulate the content of Ca2+ in the ischemic area of brain, inhibit Ca2+ overload, so as to protect neurons from ischemic injury. PMID- 12584797 TI - [Current status of development of integrated traditional Chinese and Western medicine in Qingxi University of Korea]. PMID- 12584798 TI - [Analysis of clinical effect of lifu paste in treating patients of long-term dialysis complicated with cutaneous pruritus]. PMID- 12584799 TI - [Clinical observation on treatment of climacteric hyperthyroidism with integrated medicine]. PMID- 12584801 TI - [Clinical trial and center of clinical trial]. PMID- 12584800 TI - [Clinical observation on treatment of chronic hepatitis B by large dose of ribonucleic acid plus Salviae injection]. PMID- 12584802 TI - [Exploration on Germany translation of Chinese medicine]. PMID- 12584803 TI - [Studies on molecular mechanism of TCM in preventing and treating hepatopathy]. PMID- 12584804 TI - A midwife's concerns. PMID- 12584805 TI - Marion's message. Teen moms. PMID- 12584806 TI - What are the strengths and weaknesses of the path you chose to becoming a midwife? How does the current controversy over the various pathways affect your practice, or your hopes for a practice? PMID- 12584807 TI - The essentials of midwifing midwives. PMID- 12584808 TI - Pedagogy in midwifery education. PMID- 12584809 TI - The power of independent practice. PMID- 12584810 TI - The slippery slope of professional education. PMID- 12584811 TI - A feminist approach to midwifery education. PMID- 12584812 TI - Meeting the needs of the contemporary midwifery student. PMID- 12584813 TI - A vision for midwifery in the United States. PMID- 12584814 TI - Life lessons. PMID- 12584815 TI - Supporting midwifery apprenticeship in the new millennium. PMID- 12584816 TI - A natural alternative to suturing. PMID- 12584817 TI - Midwifery and sexual misconduct. PMID- 12584818 TI - The calling. PMID- 12584819 TI - Challenging births: 2. PMID- 12584820 TI - Promoting midwifery education around the world. Interview by Kristen Graser. PMID- 12584821 TI - Italian-American women and changing childbirth practices. PMID- 12584822 TI - Watering the seeds with tears. A midwife reflects on life and death. PMID- 12584823 TI - The Jewish midwife. PMID- 12584824 TI - Mercy in action. Training missionary midwives to serve the poor. PMID- 12584825 TI - Sense and sensibility in midwifery. PMID- 12584826 TI - [Anti-liver fibrosis with integrative Chinese and Western medicine]. PMID- 12584827 TI - [Clinical study on treatment of liver fibrosis in patients of hepatitis B by kangxian baogan decoction]. AB - OBJECTIVE: To explore the clinical effect and therapeutic mechanism of Kangxian Baogan Decoction (KXBGD) on liver fibrosis caused by chronic hepatitis B. METHODS: Eight-one patients with chronic hepatitis B were divided into two groups randomly. The 54 patients in the treated group were treated by KXBGD and the 27 patients in the control group were treated by conventional liver protecting treatment. Serum levels of hyaluronic acid (HA), procollagen type III (PC III), collagen type IV (C-IV), laminin (LN), transforming growth factor beta 1 (TGF beta 1) and tumor necrosis factor-alpha (TNF-alpha) were measured before and after treatment, meanwhile, liver function and pathological changes of liver tissues were observed. RESULTS: The total effective rate in the treated group was significantly higher than that in the control group. Serum levels of HA, PC III, C-IV, LN, TGF beta 1 and TNF-alpha in the treated group obviously reduced after treatment, and the liver function got better with significant difference as compared with those in the control group (P < 0.05 or P < 0.01). Pathological examination of liver biopsy showed that the fibrous tissue in the liver reduced. CONCLUSION: KXBGD has a definite effect of anti-liver fibrosis. PMID- 12584828 TI - [Relationship between serum load of HBV-DNA and therapeutic effect of oxymatrine in patients with chronic hepatitis B]. AB - OBJECTIVE: To study the relationship between serum load level of HBV-DNA and therapeutic effect of oxymatrine in patients with chronic hepatitis B. METHODS: Forty-four patients of chronic hepatitis B were divided into two groups, the treated group was treated with oxymatrine 0.4 g/d by intramuscular injection for 3 months, the control group was treated with some liver protecting agents to estimate the therapeutic effect. The serum level of HBV-DNA was determined by quantitative polymerase chain reaction (PCR) before and after treatment. RESULTS: The seroconversion rate of HBV-DNA and HBeAg in the treated group was 43.47% and 43.47% respectively, which was obviously better than those in the control group respectively (P < 0.05), the quantity of HBV-DNA decreased after treatment from 10(6.83 +/- 1.27) copy/ml to 10(3.35 +/- 3.08) copy/ml. Among them, in patient with HBeAg negative conversion, the pretreatment quantity of HBV-DNA was 10(6.30 +/- 1.42) copy/ml, while in those with no HBeAg negative conversion, it was 10(7.23 +/- 1.23) copy/ml, the difference was significant. CONCLUSION: Oxymatrine is effective in treating chronic hepatitis B. The therapeutic effect is better for patients with lower quantity of serum HBV-DNA. PMID- 12584829 TI - [Clinical immunological study on prevention and treatment of chronic pyelonephritis with yishenkang granule]. AB - OBJECTIVE: To investigate the relationship between chronic pyelonephritis (CPN) and immune function, and the therapeutic mechanism of Yishenkang granule (YSKG). METHODS: One hundred and twenty patients of CPN were divided into 3 groups randomly, the YSKG group (treated with YSKG), the control A group (treated with Sanjin tablet) and the control B group (treated with western medicine). Serum levels of immunoglobulin (Ig), complement 3 (C3), interleukin-2 (IL-2), peripheral T-lymphocyte subsets, and urinary secretory immunoglobulin A (sIgA) were determined before and after treatment with monoclonal antibody assay, agar diffusion method, radioimmunoassay (RIA), and radioimmuno-equilibrium method, and compared with normal control. RESULTS: There were disorders of T-lymphocyte subsets in CPN, lowering of serum Ig, C3 and urinary sIgA, and increase of blood IL-2 (P < 0.05 or P < 0.01). These abnormalities could be normalized after YSKG treatment. CONCLUSION: Functional disorders of cellular and humoral immunity exist in CPN patients of chronic stage, YSKG could correct the immune functional disorder, control the recurrence of CPN effectively and alleviate the immunopathological damage. PMID- 12584830 TI - [Clinical observation on effect of shenle capsule in treating mesangial proliferating glomerulonephritis]. AB - OBJECTIVE: To study the therapeutic effect of Shenle capsule (SLC) in treating mesangial proliferating glomerulonephritis (MsPGN) and to explore its therapeutic mechanism and clinical indication. METHODS: Adopting case control method, taking angiotensin-converting enzyme inhibitor (benazepril) as control agent, the 142 cases of MsPGN were randomly divided into 3 groups, treated with SLC (Group A, n = 36), SLC plus benazepril (Group B, n = 68) and benazepril alone (Group C, n = 38) respectively. Changes of fibrinogen, lipids, renal function and urinary protein were observed. RESULTS: The total effective rate in Group A was higher than that in Group C, but with insignificant difference. The total effective rate in Group B after 3 courses of treatment was 89.74%, which was higher than that in Group C and Group A (P < 0.05). Levels of cholesterol (CH), triglyceride (TG), serum creatinine, fibrinogen and urinary protein (UP) were significantly lowered in Group A after treatment, with the levels of CH, TG and UP lower than those in Group C, while CH, TG and fibrinogen were unchanged in Group C after treatment. CONCLUSION: SLC is superior in higher efficacy and less side-effects in treating MsPGN, its effect is related with the degree of kidney pathological changes, it is more effective in treating patients with mild pathological change than in those with severe change. The outcome of combined use of SLC and angiotensin converting enzyme inhibitor would be better. PMID- 12584831 TI - [Clinical study on effect of Astragalus injection on left ventricular remodeling and left ventricular function in patients with acute myocardial infarction]. AB - OBJECTIVE: To observe the effect of Astragalus injection (AI) on left ventricular remodeling and left ventricular function in patients with acute myocardial infarction (AMI). METHODS: AMI patients were randomly divided into the AI group (54 cases) treated with AI and the control group (54 cases) treated with conventional treatment. Left ventricular end-diastolic volume index (LVEDVI), left ventricular end-systolic volume index (LVESVI), anterior endocardial segmental length (ASL), posterior endocardial segmental length (PSL) were assessed by echocardiogram at the 1st and the 4th week of treatment; and the cardiac systolic and diastolic functions were detected by nuclide gating cardiac blood pool imaging on the 4th week. Besides, the plasmic levels of lipid peroxide (MDA), count of endothelial cell (CEC) and superoxide dismutase (SOD) were determined before and after treatment. RESULTS: At the 4th week, changes of LVEDVI, LVESVI and ASL in the AI group were not obvious, but increased significantly in the control group, the significant difference in comparison between the two groups was shown (P < 0.05). As compared with the control group, in the AI group, the left ventricular ejection fraction, left ventricular peak ejecting rate and left ventricular peak filling rate were higher, and the left ventricular time for peak filling rate was shorter, moreover, MDA and CEC were lower and SOD was higher. The difference between groups and between before and after treatment were significant (P < 0.01 or P < 0.05). CONCLUSION: AI is one of the effective drugs in reversal of left ventricular remodeling and improving left ventricular function in patients with AMI. PMID- 12584833 TI - [Analysis on translation of verbs commonly used in Chinese medicine literature]. PMID- 12584832 TI - [Preliminary exploration on effect of yiqi wenyang huoxue lishui treatment on neuroendocrine system in patients with congestive heart failure]. AB - OBJECTIVE: To study the effect of Yiqi Wenyang Huoxue Lishui principle (YWHL, a therapeutic principle with Chinese medicine for reinforcing Qi warming Yang, activating blood circulation and promoting urination) on some neuroendocrine factors in patients with congestive heart failure (CHF). METHODS: Forty-nine patients of CHF with heart function of grade III-IV were randomly divided into 2 groups. The 29 cases in the treated group were treated with YWHL and the 20 cases in the control group treated with captopril. Changes of angiotensin II (AT II), atrial natriuretic peptide (ANP), endothelin (ET), nitric oxide (NO) and alpha granule membrance protein -140 (GMP-140) were observed. RESULTS: After treatment, plasma level of AT II, ANP, ET, and NO lowered in both groups with insignificant difference, but the GMP-140 reduced more significantly in the treated group (P < 0.05). CONCLUSION: YWHL showed a regulatory effect of neuroendicrine system partially similar to that of angiotensin-converting enzyme inhibitor, it possibly can improve the ventricular remodeling and would be beneficial to prevent the thrombus formation and improve heart failure by means of inhibiting platelet activity. PMID- 12584834 TI - [Clinical study on effect and therapeutical mechanism of composite Salvia injection on acute cerebral infarction]. AB - OBJECTIVE: To investigate the curative effect and therapeutical mechanism of composite salvia injection (CSI) in treating ischemic cerebral infarction in the respect of oxygen free radical and apolipoprotein. METHODS: Sixty-eight cases of ischemic cerebral infarction within the first 72 hrs after onset were divided randomly into the CSI group (treated with CSI) and the control group (treated with Xueshuantong). Serum lipid peroxide (LPO) and superoxide dismutase (SOD) were measured by colorimetry and apolipoprotein A1 (ApoA1) and ApoB100 were measured with unidirectional immune diffusion method. RESULTS: Serum levels of LPO and ApoB100 were obviously lower, and levels of SOD and ApoA1 significantly higher in the CSI group than those in the control group (P < 0.05 or P < 0.01). The total effective rate of CSI in treating cerebral infarction was 88.24%, which was significantly higher than that of the control (P < 0.05). CONCLUSION: CSI shows definite effect in treating cerebral infarction, to reduce the oxygen free radical damage and regulate the apolipoprotein metabolism possibly was the important therapeutical mechanism. PMID- 12584836 TI - [About the English translation on terms dealing with Chinese medical theory of pathogenesis]. PMID- 12584835 TI - [Effect of si' an capsule combined enalapril on hypertensive trough/peak ratio and circadian rhythm of blood pressure in patients with essential hypertension]. AB - OBJECTIVE: To investigate the effect of Si' an Capsule combined Enalapril on hypertensive trough/peak ratio (T/P) and circadian rhythm of blood pressure in patients with essential hypertension (EH). METHODS: Sixty patients with EH of stage II, III were randomly divided into two groups, the 30 patients in the treated group were treated with Si' an capsule combined Enalapril, and the 30 patients in the control group were treated with Enalapril alone. All patients were given 24-hour ambulatory blood pressure monitoring before and after 4 weeks treatment. T/P of systolic and diastolic blood pressure (SBP & DBP) of each group were calculated. Circadian rhythm of blood pressure was observed at the same time. RESULTS: T/P of SBP and DBP in the treated group was 62.4 +/- 7.0% and 66.3 +/- 8.1% respectively, which was obviously higher than that in the control group, 53.3 +/- 6.7% and 60.1 +/- 7.2%, respectively (P < 0.05). The blood pressure circadian rhythm recovering rate in the treated group was 73.3% and in the control group 50%, the difference was insignificant. CONCLUSION: The combination therapy of Si' an capsule and Enalapril could lower the blood pressure smoothly and restore the circadian rhythm of blood pressure in EH patients. PMID- 12584837 TI - [Study on abnormality and regulation of T-lymphocyte subsets in peripheral blood of rheumatoid arthritis patients]. AB - OBJECTIVE: To investigate the change of T-lymphocyte subsets in peripheral blood of rheumatoid arthritis (RA) patients and analyze the effects of Fuzheng Qubi Decoction (FZQBD) on T-cell subsets. METHODS: Thirty RA patients were randomly divided into two groups, and treated with FZQBD or western medicine combination therapy respectively for one month, the percentage of peripheral CD3+, CD4+, CD8+ as well as the serum IgG and IgA levels were determined. RESULTS: The percentage of peripheral CD4+, CD4+/CD8+ ratio, IgG and IgA levels increased significantly (P < 0.001) in RA patients before treatment. After 1 month of FZQBD treatment, the CD4+/CD8+ ratio decreased obviously (P < 0.05). CONCLUSION: Abnormal cellular immunity exists in RA patients, FZQBD could adjust the abnormal T-lymphocyte subsets to normalize it. PMID- 12584838 TI - [Effect of herbal compound 861 on expression and activity of nitric oxide synthase in hepatic stellate cells]. AB - OBJECTIVE: To study the effect of herbal compound 861 (HB861) on expression and activity of nitric oxide synthase (NOS) in hepatic stellate cells (HSC), and to explore the feasibility of its application in preventing and treating the early portal hypertension. METHODS: HSC of HSC-T6 cell line (1 x 10(5)/ml) were cultured in dish with 95% O2 plus 5% CO2 under 37 degrees C for 24 hrs, then divided into 5 groups, 6 dishes in each group. Group A was the blank control group. To Group B-E, HB861 2 mg/ml, HB861 4 mg/ml, HB861 8 mg/ml, HB861 4 mg/ml + NW-Nitro-L-Arginine Methyl Ester (L-NAME)4 mg/ml were added separately, and continuously cultured for 24 hrs. NOS activity was measured using colorimetry, NO level was determined by nitrate reductase technique. The cells were fixed by 4% paraformaldehyde for 2 hrs for test HSC-T6 iNOS expression by immunocyto-chemical method. RESULTS: HB861 in 2 mg/ml, 4 mg/ml and 8 mg/ml could increase HSC-T6 NOS activity from 1.7 +/- 0.1 to 2.5 +/- 0.3, 3.5 +/- 0.4 and 3.7 +/- 0.9 respectively (P < 0.01), the NO levels in supernatant were increased in parallel from 56.1 +/- 4.8 to 90.7 +/- 4.6, 99.7 +/- 4.1 and 109.0 +/- 2.7 respectively (P < 0.01). L-NAME could not inhibit the effect of HB861 in increasing the synthesis and secretion of NO by activated HSC-T6. Immuno-cyto-chemical study showed that there was iNOS expression in cytoplasm, and which could be increased by HB861. CONCLUSION: The activated HSC-T6 showed positive iNOS expression, suggesting it could produce NO. HB861 could markedly increase HSC-T6 iNOS expression and NOS activity, enhance the NO synthesis and secretion, it also could inhibit the contractility of activated HSC by way of increase HSC to secrete NO, so as to lower the resistance in hepatic sinusoid, therefore would play important role in preventing and treating of early portal hypertension. PMID- 12584839 TI - [Effect of ligustrazine on bone marrow hematopoiesis in mice after bone marrow transplantation]. AB - OBJECTIVE: To investigate the effect of ligustrazine (LT) on hematopoiesis in mice after bone marrow isotransplantation (iso-BMT). METHODS: The typical model of iso-BMT was established and the model mice were randomly divided into two groups, the LT group treated with LT injection 0.2 ml and the control group treated with normal saline 0.2 ml, twice a day by gastrogavage. The following parameters were observed in the day 1, 7 and 14: peripheral blood cells, bone marrow mono-nuclear cells (BMMNC), heparin sulfate (HS) expression in bone marrow section by immunohistochemical SABC-AP method, stromal cell derived factor-1 (SDF 1) expression and CXC chemotaxis factor receptor 4 (CXCR4) expression. RESULTS: The levels of peripheral WBC, platelet, BMMNC, CXCR4, HS, SDF-1 at the day 7 and 14 in the LT group were all higher significantly than those in the control group (P < 0.05 or P < 0.01). CONCLUSION: LT could improve the bone marrow hematopoiesis in the early hematopoietic re-establishing stage after BMT. PMID- 12584840 TI - [Study on preventive effect of jiangu granule on osteoporosis in ovariectomized rats]. AB - OBJECTIVE: To investigate the preventive effect of Jiangu granule (JGG) on experimental primary osteoporosis type I. METHODS: Osteoporosis model was established through ovary resection of female rats. Bone mineral density (BMD) was measured with double energy X-ray absorptiometry. Level of endocrine markers, including osteocalcin (BGP), estradiol (E2) and calcitonin (CT) in serum were examined by RIA. And uterus parameters was calculated also. RESULTS: JGG could significantly increase BMD and uterus index, improve the levels of E2 and CT in serum, at the same time reduce the BGP level. CONCLUSION: JGG can efficiently prevent type I primary osteoporosis or delay its occurrence by enhancing the function of endocrine system, coordinating the action of calcium related hormone, reducing bone turnover rate and increasing BMD. PMID- 12584841 TI - [Effect of lupus recipe on IL-6 and IL-10 secretion of splenic cells in vitro in lupoid mice]. AB - OBJECTIVE: To investigate the effect of Lupus Recipe (LR) on IL-6 and IL-10 secretion of splenic cells in vitro in lupoid model mice and on anti-dsDNA antibody. METHODS: Chronic graft-versus-host disease model was used in the experiment. The model mice were divided into four groups, the model group was un treated and the other three groups treated with LR, prednisone and combined treatment (prednisone + LR) respectively. The serum level of ds-DNA antibody, the ConA induced splenic cell proliferation in mice's splenic cell culture as well as the IL-6, IL-10 level in the supernatant of culture were determined after treatment and compared with those of normal controls. RESULTS: (1) The splenic cell proliferative reaction in the model group splenic cells was obviously higher than that of the normal control (P < 0.05); but that in the three treated groups was different from the control insignificantly (P > 0.05); (2) The serum anti dsDNA in the model group was higher than that in the normal control, 1.75 +/- 0.25 vs 1.20 +/- 0.21 (P < 0.01), while the difference in comparison of the treated groups with the normal control was insignificant, (P > 0.05); (3) Splenic cell IL-6 and IL-10 secretion in the model group induced by ConA was higher than those in the treated groups and the controls significantly (P < 0.05). CONCLUSION: LR reveals the effect of immunosuppressor, which could inhibit the activation of T- and B-cells, reduce the Th2 cytokine formation and auto-antibody production so as to treat lupus erythematosus effectively. PMID- 12584842 TI - [Comparative study on effect of recipe for activating blood circulation and replenishing Qi on left ventricular remodeling in rats with left heart failure after myocardial infarction]. AB - OBJECTIVE: Comparative study to the effect of Chinese herbal medicine on left ventricular remodeling in rats with left heart failure after myocardial infarction (MI). METHODS: Rat's model of left heart failure after myocardial infarction was treated with injection for activating blood circulation (ABCI, consisted of R. Salviae miltiorrhizea; Rh. Ligusticum wallichii and F1. Carthamus tinctorius) and injection for replenishing Qi (RQI, consisted of R. Codonopsis Pilosulae and R. Astragalus membranaceus) respectively. The effect of treatment were evaluated by observing and comparing the changes of heart morphological structure, collagen element, heart weight/body weight ratio (HW/BW), left intraventricular area (LVA), ratio of ventricular wall thinning in MI area and myocardial nuclei number (MNN) per square area. RESULTS: In comparison with the model group, the reduction of collagen tissue around myocardial cells in living area of MI, HW/BW and LVA of ABCI and RQI group were lower, and MNN per square area was higher significantly (all P < 0.05). CONCLUSION: Both ABCI and RQI, though without positive myodynamia, showed certain inhibitory effect of left ventricular remodeling in rats with left heart failure after MI. PMID- 12584843 TI - [Effect of Chinese bushen huatan drugs on pituitary-sex gland-immune axis in smoking induced chronic bronchitis mice]. AB - OBJECTIVE: To study the effect of Chinese Bushen Huatan Drugs (BSHT, Chinese drugs for Kidney tonifying and phlegm resolving) on pituitary-sex gland-immune axis in smoking induced mice model of chronic bronchitis. METHODS: Model mice were induced by smoking and divided into the model control group, the large dosage of BSHT group, the small dosage of BSHT group and the positive control group (treated by Guilong Kechuanning, a proved effective Chinese patent drug). A blank group of normal mice was also set for control. Serum levels of testosterone (T), luteotropic hormone (LH), interleukin-2 (IL-2), interleukin-8 (IL-8), visceral indexes of testis, epididymis and thymus were measured and microstructure of testis tissue was also observed. RESULTS: As compared with those in the blank group, serum T, LH and IL-2 levels, and testis, epididymis and thymus indexes were all lower, and IL-8 level was higher in the model control group, moreover, atrophic change of testis was present in the model mice. These abnormal changes were all improved in the BSHT groups. CONCLUSION: Smoking induced mice model of chronic bronchitis, which may cause reproductive endocrine disturbance and immunosuppression. BSHT could modulate the pituitary-sex gland immune axis through adjusting the disturbed sex hormone, improve the pathological change of testis and enhance the immunity of organism. PMID- 12584844 TI - [Effect of compound Salvia pill combined with propanolol on liver fibrosis and portal hypertension]. PMID- 12584845 TI - [Clinical observation on effect of heze oral liquid in treating children simple obesity]. PMID- 12584846 TI - [Juanbi mixture in treating 100 cases of rheumatoid arthritis patients]. PMID- 12584847 TI - [Material basis studies of Chinese drugs]. PMID- 12584848 TI - [Study on singly Chinese herbal drugs in intervention of visceral fibrosis]. PMID- 12584849 TI - [The minutes of the fifth national integrative Chinese and Western medicine blood stasis syndrome and activating blood circulation to remove stasis conference research]. PMID- 12584850 TI - [Effects of 18 alpha-glycyrrhizic acid on rat liver cytochrome P450 isoenzymes and phase II transferase]. AB - AIM: To study the effect of 18 alpha-glycyrrhizic acid (18 alpha-GL) on hepatic microsomal drug metabolizing enzymes in rats. METHODS: 18 alpha-GL (12.5, 50.0 mg.kg-1.d-1) were given i.p. to male Wistar rats for 3, 6 or 12 consecutive days. The rats were sacrificed 24 h after the last dose and the liver microsomes were prepared for analysis of cytochrome P450 (CYP) isozymes and phase II transferase activites. RESULTS: Aniline hydroxylase (CYP2E1) activities in the rats treated with 18 alpha-GL (12.5, 50.0 mg.kg-1) for 6 days decreased dose-dependently by up to 53.2%; For 3, 6 or 12 days 7-ethoxyresorufin O-deethylase (CYP1A1) activities in the rats of 50 mg.kg-1 dose group decreased time-dependently by 17.6%, 38.3% and 47.3%, respectively; Erythromycin N-demethylase (CYP3A) activities was significantly inhibited from 23.1% to 34.3%. UDP-glucuronosyltransferase activities toward 7-hydroxy-4-methylcoumarin significantly increased ranging from 19.3% to 29.9%. UDP-glucuronosyltransferase activities toward 4-phenylphenol in the rats treated with 18 alpha-GL (12.5, 50.0 mg.kg-1) for 6 days increased by 45.9% and 70.3%. Glutathione S-transferase (GST) activities in the rats treated with 18 alpha-GL (12.5, 50.0 mg.kg-1) for 6 days increased by 13.7% and 48.3% in dose-dependent manner. CONCLUSION: 18 alpha-GL inhibited rat liver microsomal cytochrome P450 while induced phase II transferase. PMID- 12584851 TI - [Pharmacokinetics of site-specific delivery of dexamethasone-dextran prodrug in rat gastrointestinal tract]. AB - AIM: To explore whether dexamethasone-dextran (260,000) has the characteristics of site-specific delivery in rat gastrointestinal tract. METHODS: Dexamethasone prodrug and dexamethasone were administered to rat ig at the dose of 5 mumol.kg 1. The distribution of dexamethasone in the contents and mucosa of different parts of the rat GI tract at different time intervals and its concentration in plasma were determined by HPLC. RESULTS: Dexamethasone was mainly released in the cecum and colon contents and mucosa after oral administration of dexamethasone prodrug. The absorption was reduced significantly. The peak time of the drug in plasma was 8.1 h, and the peak concentration was 32 micrograms.L-1. However, free dexamethasone was found mainly in the contents and mucosa of the stomach, proximal and distal small intestine. The peak time of the drug in plasma was 2.2 h, and the peak concentration was 2120 micrograms.L-1. CONCLUSION: Dexamethasone can be specifically delivered to the large intestine by using dexamethasone dextran (260,000). It appears that the prodrug has a potential in the treatment of inflammatory bowel disease. PMID- 12584852 TI - [Effects of 3-n-butylphthalide on thrombosis formation and platelet function in rats]. AB - AIM: To study the effects of dl-, l- and d-3-n-butylphthalide (NBP) on platelet aggregation and thrombus formation. METHODS: Thrombus formation was assessed by silk thread-induced thrombosis in arteriovenous shunt in rats. Rat platelet aggregation induced by adenosine diphosphate (ADP), arachidonic acid (AA), collagen and thrombin was detected in vitro. The generation of thromboxane B2 (TXB2) and the concentration of cAMP in rabbit platelets in vitro were studied with radioimmunological assay. RESULTS: dl-, l-NBP (5, 10, 20 mg.kg-1 i.p.) exhibited a dose-dependent inhibitory effect on thrombus formation in rats, while d-NBP was not active. dl, l, d-NBP (3-100 mumol.L-1) inhibited platelet-rich plasma aggregation in vitro induced by ADP, collagen and AA, and all of them showed no effect on thrombin-induced platelet aggregation. In addition, dl, l-NBP (10-100 mumol.L-1) were found to increase [cAMP]i in dose-related fashion. In the meantime, only high concentration of l-NBP was found to decrease platelet TXA2 level. In addition, l-NBP (1-100 mumol.L-1) showed significant effect on inhibiting 5-HT release from platelets. In contrast, dl- and d-NBP showed no effect. CONCLUSION: The results suggest that NBP is a potent antiplatelet drug, the mechanism of its antithrombotic and antiplatelet activity is related to its regulation of cAMP level and 5-HT release. PMID- 12584853 TI - [Rhein induces apoptosis in cancer cells and shows synergy with mitomycin]. AB - AIM: To study the apoptosis-inducing and growth-inhibitory effect of rhein, an herb-derived compound, and its combination with mitomycin C (MMC) on cultured tumor cells. METHODS: MTT assays were used to determine the inhibition of proliferation by drugs in several tumor cell lines. Nucleoside transport and DNA synthesis inhibition were determined by [3H] thymidine transport and incorporation assays. Flow cytometry, electrophoresis on agarose gels and morphological assessment were applied to analyze the apoptotic changes. RESULTS: The IC50 values of nucleoside transport was 19.1 micrograms.mL-1 and that of the DNA synthesis inhibited by rhein was 27.4 micrograms.mL-1. In MTT assay the IC50 values of rhein for KB, hepatoma BEL-7402 and mammary carcinoma MCF-7 cells were 11.5 micrograms.mL-1, 14.0 micrograms.mL-1 and 18.4 micrograms.mL-1 respectively. Synergistic effect of rhein and MMC was found in all the three cell lines. As found, rhein induced apoptosis in KB cells, and the increase of apoptotic cells reached 71.0% at 96 h. The combination of rhein and MMC enhanced the induction of apoptosis significantly. CONCLUSION: These results suggest that rhein, as a biochemical modulator, might be potentially useful in cancer chemotherapy. PMID- 12584855 TI - [3D-QSAR of antiinflammatory activities of 5,6-diaryl-2,3-dihydro-1-pyrrolizinone derivatives]. AB - AIM: To study the SARs of 5,6-diaryl-2,3-dihydrol-1-pyrrolizinone derivatives to provide information for the design of new structural compounds. METHODS AND RESULTS: Three dimensional quantitative structure-activity relationship (3D-QSAR) model was constructed by Apex-3D. CONCLUSION: The antiinflammatory activities of 5, 6-diaryl-pyrrolizinones were related to the global hydrophobicity and volume, the properties of the group at 1-position of pyrrolizinone ring and the two secondary sites; improved the pi-electronic density of the group at 1-position of pyrrolizinone ring and lowered the global hydrophobicity and the volume of p substituent of the phenyl ring at 6-position of pyrrolizinone contributed to the antiinflammatory activities of the title compounds. PMID- 12584854 TI - [Effects of fluid percussion injury on intracellular [Ca2+]i and pH in cultured rat neurons]. AB - AIM: To study the change of intracellular [Ca2+]i and pH in cultured neurons after fluid percussion injury, and the therapeutic effect of drugs. METHODS: The neurons of Sprague Dawley rats were cultured for 8-14 days, then treated them with fluid percussion injury (2.5 kPa, 20 ms). Alterations of [Ca2+]i and pH in single neural cells following fluid percussion injury were measured by a laser scanning confocal microscope. After being injured for several hours the cultured neurons were treated with nimodipine or D-(-)-2-amino-5-phosphonovaleric acid (D AP-5). Two hours later, the effects of drugs on intracellular [Ca2+]i and pH were studied. RESULTS: The Intracellular [Ca2+]i increased quickly after brain injury and reached peak in 12 hours. It then decreased gradually and became normal at 48 hours. The pH decreased slowly, reached minimum in 12 hours, and then kept at a lower level. It did not recover normal at 48 hours. Nimodipine and D-AP-5 decreased significantly the ascension of [Ca2+]i and the descent of pH. But nimodipine and D-AP-5 must be given within 10 hours after injury for a good therapeutic effect. CONCLUSION: According to the change of intracellular [Ca2+]i and pH, early use of nimodipine and D-AP-5, will get a better therapeutic effect. PMID- 12584856 TI - [Three dimensional quantitative structure-activity relationship of heterocyclic compounds with di-tert-bultylphenyl inhibitors]. AB - AIM: To quatitatively disclose the relationship between activity and structure of a new class of COX-II inhibitors containing dialkylphenyl-linked heterocyclic moieties. METHODS AND RESULTS: Seventeen COX-II inhibitors from literature as a training set were investigated with the aim of developing a 3D-QSAR model using comparative molecular field analysis (CoMFA). To reveal the pharmacophoric pattern, several modes of superimposition were explored. The significant model shows a higher ability to explain and predict the activity of COX-II inhibitors, with the cross-validation RCV2 = 0.718, non cross-validation R2 = 0.992, F = 260,624, and SEE (standard error of estimate) = 0.072. Three compounds were selected as a predicting set, the low deviations of calculated values from the measured ones suggesting a powerful predictive ability of the model. CONCLUSION: The 3D-QSAR explains the dependence of COX-2 inhibition upon the structures of the compounds. Some structure information for design of new COX-II inhibitors with higher activity has been given. PMID- 12584857 TI - [Chemical constituents of Peucedanum decursivum]. AB - AIM: To study the chemical constituents of Peucedanum decursivum. METHODS: Various column chromatographies with silica gel and HPLC were employed for the isolation and purification. The structures of the compounds were elucidated on basis of spectral analyses and chemical methods. RESULTS: Seven compounds were isolated from the roots of Peucedanum decursivum (Miq.) Maxim. 1 is 3'(S)-hydroxy 4' (R)-angeloyloxy-3',4'-dihydroxanthyletin, named Decursitin D; 2 is 3'(S) acetoxy-4'(R)-hydroxy-3',4'-dihydroxanthyletin, named decurstin F; the others are 3'(S)-acetoxy-4'(R)-angeloyloxy-3', 4'-dihydroxanthyletin (3), Pd-C-IV (4), Pd-C II (5), (+)-3'S-decursinol (6) and (+)-trans-decursidinol (7). CONCLUSION: Compounds 1 and 2 are new compounds, while compounds 6 and 7 were isolated from Peucedanum decursivum (Miq.) Maxim for the first time. PMID- 12584858 TI - [Structure elucidation of glycan of a glycoconjugate SPPA-1 isolated from Spirulina platensis]. AB - AIM: To elucidate the structure of the glycan of SPPA-1, a glycoconjugate isolated from Spirulina platensis. METHODS: Methylation analysis, GC/MS, and 1D, 2DNMR techniques were used to determine the structures of the glycoconjugate (SPPA-1). RESULTS: SPPA-1 was only composed of alpha-D-glucose and shown to be a (1-->4) linked alpha-D-glucan to which a few glucosyl side chains are attached at O-6 of the glucosyl residues of the main chain. CONCLUSION: The glycan of SPPA-1 is a new glucan. PMID- 12584859 TI - [Study on the structure and activity of new phenolic acid compounds from Erigeron breviscapus]. AB - AIM: To isolate and identify the active constituents of Erigeron brevisapus (Vant.) Hand.-Mazz. METHODS: The chemical constituents were isolated by silica gel column chromatography and two new compounds were obtained. Their structures were elucidated by IR, MS, 1HNMR, 13CNMR, DEPT and 2DNMR. The injury of BCMEC (bovine cerebral microvascular endothelial cell) was determined by lactic dehydrogenase (LDH), the ability of the drugs anti-oxidation and scavenging oxidation of free radical was measured by colorimetric method. RESULTS: Two new compounds have been identified as 1-O-methyl-3, 5-O-dicaffeoyl quinic acid methyl ester (III) and 5-O-caffeoyl quinic acid butyl ester (IV). CONCLUSION: Compounds III and IV are new compounds. Compound III can protect BCMEC injury by LPC. PMID- 12584860 TI - [Studies on in-vivo process of the enantiomers of nicardipine in rabbit by coupled chiral and achiral HPLC]. AB - AIM: To study the differences of the pharmacokinetics and tissue distribution of two enantiomers of nicardipine in the rabbit. METHODS: Biological samples were diluted by 1 mol.L-1 NaOH solution and extracted with n-hexane--ethyl acetate (1:1). The concentrations of S-nicardipine and R-nicardipine in samples were determined by coupled achiral C18 column (150 mm x 4.6 mm, 5 microns) and chiral OJ column (250 mm x 4.6 mm, 10 microns) chromatography. RESULTS: The racemic nicardipine and the enantiomers in sample were separated well by the coupled system. The linear range was 55-550 ng.mL-1 for both enantiomers. The within-day and between-days RSD (n = 5) were 5.25% and 8.97%, and the relative recoveries were 99.99% and 97.10% for R- and S-enantiomer, respectively. The mean Tmax, Cmax and AUC values were (2.49 +/- 0.03) h, (134 +/- 2) ng.mL-1 and (1082 +/- 32) ng.mL-1.h for S-nicardipine and (1.24 +/- 0.05) h, (109 +/- 2) ng.mL-1 and (778 +/- 22) ng.mL-1.h for R-nicardipine. The concentration of S-nicardipine were generally higher than that of R-nicardipine in main target tissues and cells. CONCLUSION: There were significant differences between the two enantiomers of nicardipine in rabbit in pharmacokinetics and tissue distribution. PMID- 12584861 TI - [Bioequivalence of bambuteral and its metabolites terbutaline after oral bambuteral tablet in healthy volunteers by HPLC/MS]. AB - AIM: To study bioequivalence of bambuteral and its metablites terbutaline in 20 healthy male volunteers. METHODS: A single oral dose of domestic or imported bambuteral tablet was given according to a randomized 2-way cross-over design. The plasma bambuteral and terbutaline concentrations were determined by HPLC/MS. RESULTS: The pharmacokinetic parameters of domestic and imported bambuteral: AUC0 t were (52 +/- 21) microgram.h.L-1 and (51 +/- 20) microgram.h.L-1, Tmax were (2.9 +/- 0.9) h and (2.6 +/- 0.7) h, Cmax were (6.0 +/- 2.6) microgram.L-1 and (6.2 +/- 2.9) microgram.L-1, T1/2Ke were (11.2 +/- 2.3) h and (11.2 +/- 1.9) h, respectively; terbutaline: AUC0-t were (191 +/- 30) microgram.h.L-1 and (197 +/- 37) microgram.h.L-1; Tmax were (4.2 +/- 1.0) h and (4.2 +/- 1.0) h; Cmax were (10 +/- 5) microgram.L-1 and (10 +/- 4) microgram.L-1; T1/2Ke were (20 +/- 3) h and (21 +/- 4) h, respectively. The bioavaiability of the domestics was 102% +/- 8% (bambuteral) and 100% +/- 12% (terbutaline). CONCLUSION: The results demonstrate that the two forms of bambuteral and terbutaline were bioequivalent by analysis of variance, two-one sided test and 90% confidential limit. PMID- 12584862 TI - [Study on the structure of epimerides of cycloclausenamide]. AB - AIM: To study the structures of the epimerides of cycloclausenamide. METHODS: The structures of compound I, extracted from Clausena lansium (Lour.) Skeels, and synthesized compound III were determined by single crystal X-ray diffraction analysis. The stereo-structures of compound II and IV were also built up through Tripos force field based on crystal structures of compound I and III. RESULTS: The molecular formula and molecular weight were found to be C18H17O2N and 279.34 respectively. Compound I crystallized in monoclinic system, space group P2(1) with a = 0.5928(1), b = 1.5014(1), c = 1.6190(1) nm, V = 1.4410(3) nm3, Z = 4, Dx = 1.288 g.cm-3, Rf = 0.075, Rw = 0.073(w = 1/sigma 2|F|), S = 3.983; compound III crystallized in triclinic system, space group P1 with a = 0.5667(1), b = 1.2934(1), c = 2.1119 (1) nm, alpha = 102.17(1), beta = 90.25(1), gamma = 102.65(2) degrees, V = 1.4770(5) nm3, Z = 4, Dx = 1.224 g.cm-3, Rf = 0.047, Rw = 0.051(w = 1/sigma 2|F|), S = 0.467. CONCLUSION: These results showed that compound I and III both are cycloclausenamide except that the directions of the phenyl group on C6 are different. Cycloclausenamide can form 4 pairs of epimerides but the directions of the phenyl group does not affect their energy in free state. PMID- 12584863 TI - [Improving bioavailability of naftopidil by using bioadhesion in dogs]. AB - AIM: To prepare naftopidil bioadhesive sustained-release capsule and study their pharmacokinetics and relative bioavailability in the dog. METHODS: Bioadhesive polymers such as hydroxypropyl methylcellulse (HPMC) and Carbopol 934 (CP 934) were used in capsule prescriptions. Naftopidil capsule and two formulations of bioadhesive sustained-release capsules (I and II) were given to five healthy male dogs in a cross-over test. The naftopidil concentrations in plasma were determined by a newly developed HPLC method and the pharmacokinetic parameters as well as the relative bioavailability were measured. RESULTS: The C0-->infinity, Cmax and Tmax of naftopidil capsule was (3728 +/- 573) h.ng.mL-1, (697 +/- 94) ng.mL-1 and (1.2 +/- 0.5) h. These parameters of bioadhesive sustained-release capsule I and II, respectively, were (5518 +/- 391) h.ng.mL-1 and (5636 +/- 427) h.ng.mL-1; (468 +/- 61) ng.mL-1 and (512 +/- 72) ng.mL-1; both (4.0 +/- 0.7) h. Results from statistics showed that there were significant difference between bioadhesive formulations and the non-bioadhesive one in C0-->infinity, Cmax and Tmax. The bioadhesive formulations and the non-bioadhesive one were not bioequivalent, the relative bioavailability of the two bioadhesive sustained release capsules were respectively 150% +/- 14% and 154% +/- 23% when compared with the non-bioadhesive capsule. CONCLUSION: It is much improving bioavailability of naftopidil by using bioadhesion. PMID- 12584864 TI - [Transdermal delivery of cyclosporin A solubilized in mixed micelles through mice skin]. AB - AIM: To investigate the transdermal delivery effects of cyclosporine A solubilized in mixed micelles composed of phospholipid and different surfactants. METHODS: When applied onto the excised abdominal skin of the mice occlusively, the enhancing effects of various mixed micelles on the penetration of cyclosporin A were assessed by an in vitro permeation technique. In vivo study was carried out by topical application of sodium cholate-phospholipid mixed micelles onto the mice skin and drug blood concentration was detected. RESULTS: In vitro, mixed micelles containing different surfactants displayed distinct permeability and corresponded to the following order: sodium cholate > sodium deoxycholate > Trition X-100 > Tween-20. In vivo, peak drug concentration was detected at 5 h and after that the concentration fell down slowly. CONCLUSION: Mixed micelles were shown to be efficient carrier for the transdermal delivery of the lipophilic polypeptide when kept in solution during the application process. PMID- 12584865 TI - [Research on DNA molecular marker of Magnolia officinalis Rehd. et Wils.--RAPD study on certified species]. AB - AIM: To discuss the intraspecific relationship in Magnolia officinalis and the genuineness of Cortex Magnoliae officinalis, and to find some DNA characters of certified "Houpo". METHODS: Thirty-three samples from eleven locations, which can represent most of the distribution of M. officinalis, were selected. The total DNA was extracted. Severty-four random primers were tried to get good amplification. RESULTS: One hundred and sixteen bands amplified from seventeen primers, were clustered by NTSYS-pc software. Three branches were obtained. Some distinctive primers and bands, which represent certified species or fine breed, were obtained also. CONCLUSION: 1) M. officinalis should be divided into three geographic clans instead of two subspecies or varieties, they are, a) typical officinalis, b) typical biloba and c) Middle type. This conclusion agrees with the leaf form and other characters. 2) The genetic difference between "Chuanpo" and "Wenpo" is evident and the difference is in correspondence with the quantities of their chemical constituents. So, the genetic difference is the main reason of the genuineness of Cortex Magnoliae officinalis. 3) These results may be used to establish DNA database for identification of Cortex Magnoliae officinalis. PMID- 12584866 TI - [Application of nuclear medicine techniques in drug development]. PMID- 12584867 TI - [Harnessing the microbial genome to develop novel antibiotics]. PMID- 12584868 TI - Clinical characteristics of under-reporters on urine drug screens in a cocaine treatment study. AB - To explore the concordance between urine drug screen (UDS) results and self report of cocaine use, results in a pharmacologic treatment trial for cocaine dependence were evaluated. Subjects with at least two occurrences of a positive UDS for cocaine were characterized as either an under-reporter (UR, n = 43) or a truthful reporter (TR, n = 32). Interestingly, URs attended more study sessions and were more likely to complete the study. Significant differences were found in cocaine use patterns and the prevalence of Axis I and Axis II disorders. This information may be important in guiding the judicious use of UDS in clinical and research settings. PMID- 12584869 TI - Need for medical and psychosocial services among injection drug users: a comparative study of needle exchange and methadone maintenance. AB - This study compares the prevalence of perceived and unmet needs of HIV-negative injection drug users (IDUs) not receiving drug treatment (n = 251) and those recruited from a methadone maintenance program (n = 312) in 1998. We studied self reported needs for six community services: medical, mental health, housing, income assistance, alcohol treatment, and drug treatment. Respondents reported the highest levels of need for mental health and housing services. Ninety-four percent of out-of-treatment IDUs reported having at least one need compared to 62% of methadone clients (p < .001). Across all reported service needs, at least 69% of respondents in both cohorts reported their needs were unmet. While HIV infected drug users receive assistance through the Ryan White CARE Act, these findings suggest that seronegative drug users may benefit from similar community service programs. PMID- 12584870 TI - Modified directly observed therapy (MDOT) for injection drug users with HIV disease. AB - Injection drug use is an important factor in the spread of HIV infection, and strategies to enhance adherence to HIV therapeutics are critically important to controlling viral transmission and improving clinical outcomes. To this end, the authors sought (1) to enhance adherence to highly active antiretroviral therapy (HAART) among methadone-maintained injection drug users (IDUs) using modified directly observed therapy (MDOT), and (2) to define interactions between methadone and HAART and the potential contribution of drug interactions to adherence and HIV outcomes in this population. Adherence was explored here through a pilot, unblinded, 24-week study in a methadone maintenance program in which simplified HAART (efavirenz and didanosine [one daily] and a second nucleoside [twice daily]) was administered 6 days/week by clinic staff to HIV infected IDUs (n = 5) with their methadone. Evening doses of riboflavin-tagged nucleoside and one full day of medication weekly were given as take home doses. As a result of HAART administration, four of five participants with mean viral load at baseline of 10(5) copies/ml had undetectable viral load by 8 weeks of treatment (p = .043). Methadone area under the curve (AUC) decreased by 55% (p = .007) within 2 weeks of initiating this HAART regimen, and a mean methadone dose increase of 52% was required. The authors conclude that MDOT is a promising intervention for the treatment of IDUs with HIV disease, though significant drug interactions must be monitored for carefully and rapidly addressed. PMID- 12584871 TI - Effectiveness of high versus low structure individual counseling for substance abuse. AB - Outcomes during treatment were compared for inner-city drug-dependent individuals randomly assigned to high-structure, behaviorally-oriented (HSB) or to low structure, facilitative (LSF) individual counseling. No group differences were found for symptom reduction, patient and counselor ratings of benefit, retention, or drug free urines. However, the more depressed and "helpless" clients showed significantly better control of substance use in HSB counseling, while the less depressed and helpless patients realized better control in LSF treatment. These results suggest that outcomes might be improved by matching patients to high- and low-structure counseling based on their pretreatment levels of depression and learned helplessness. PMID- 12584872 TI - Client satisfaction with three therapies in the treatment of alcohol dependence: results from project MATCH. AB - This study investigates client satisfaction with the three therapies for the treatment of alcohol dependence used in Project MATCH. Primary findings indicate that (1) clients receiving these therapies as aftercare were more satisfied than those receiving them as outpatients; (2) outpatients were less satisfied with Motivational Enhancement and Twelve-Step therapies than with Cognitive-Behavioral therapy; and (3) higher levels of satisfaction were associated with higher levels of therapy attendance, greater reductions in drinking during therapy, and better clinical status at the end of therapy. The results reflect the complexity of the relationship among treatment process, therapy outcome, and client satisfaction. PMID- 12584873 TI - Effects of HIV triple therapy on methadone levels. AB - There is a belief among methadone patients that triple therapy for HIV reduces methadone potency. This cross-sectional study compared the rate of methadone metabolism (peak-trough blood levels) in two groups of methadone-maintained patients, AIDS patients receiving triple therapy (N = 17), and HIV patients without triple therapy (N = 19). These preliminary findings suggest that triple therapy may increase the rate of methadone metabolism, though further studies are warranted. PMID- 12584874 TI - Naturalistic, self-assignment comparative trial of bupropion SR, a nicotine patch, or both for smoking cessation treatment in primary care. AB - The effectiveness of bupropion SR, a nicotine patch, or both, when co administered with cognitive-behavioral therapy, on smoking cessation was evaluated under treatment-as-usual conditions in a primary care smoking cessation clinic. Consecutive consenting patients (n = 189) assigned themselves to one of three treatments according to personal preference: nicotine patch (NTP; n = 27), bupropion SR (B; n = 101), and bupropion SR plus nicotine patch (B + NTP; n = 61). Six-month self-reported abstinence rates were 14.8%, 27.7%, and 34.4%, respectively. Odds ratios for 6-month abstinence, with NTP as reference, were 2.2 (CI = 0.7-5.9; p = .19) for B, and 3.0 (CI = 0.9-9.9; p = .07) for B + NTP. Resemblance of abstinence pattern and magnitude between this naturalistic study and randomized clinical trials of bupropion SR may support tentative inferences of stability and generalizability of clinical trial efficacy findings. PMID- 12584875 TI - Ketamine as a preferred substance of abuse. PMID- 12584876 TI - [Meniere's disease: diagnostic criteria, criteria to establish stages, and standards for treatment evaluation. Bibliographic review and update]. AB - If all doctors would use the same concepts to define Meniere's Disease (MD) and the same scale to either establish the different steps of the disease or to evaluate a specific treatment, it would be easy to compare our results. This would be beneficial for patients because of the possibility of giving them a more accurate information about the follow-up of their disease and the advantages of the different possibilities of treatment. Since the introduction of informatic systems a new goal appeared: large databases could be achieved, as a result of being able to gather the different database that are independently used in each hospital, provided that rules to establish data would be previously standardized. The purpose of this paper is to evaluate the proposal of the Committee on Hearing and Equilibrium of the American Academy of Otolaryngology and those proposed by other authors according to the stated courses of action. PMID- 12584877 TI - [Subcutaneous autograft with newly synthesized cartilage using ethisorb polymer in rabbits]. AB - We perform a subcutaneous autograft, in animals with preserved immunity (24 rabbits), of cartilage taken from the auricle, treated with tissue engineering thecnics and embeded in a reabsorbable polimer (Ethisorb) that acts as base. We observed a good quality cartilage with the expression of collagen type II and without graft rejection phenomenon. PMID- 12584878 TI - [Audiometric features of familial hearing impairment transmitted by mitochondrial inheritance (A1555G)]. AB - OBJECTIVE: To examine the audiometric patterns of familial hearing impairment due to the A1555G mutation in the mitochondrial DNA. PATIENTS AND METHODS: We include 55 subjects with the A1555G mutation from 6 unrelated families, affected by nonsyndromic sensorineural hearing loss and residing in Cantabria. The A1555G mutation was found in homoplasmy in all the families, except in one family, in which it was in heteroplasmy. Aside from standard history taking and general otolaryngological examination, pure tone audiometry was carried out in all patients. RESULTS: Hearing loss was developed by most of the patients. The auditory defect was a slowly progressive bilateral symmetrical sensorineural hearing loss, affecting mainly the high frequencies. In patients in which aminoglycoside ototoxicity could be excluded, hearing loss usually ranged from mild to moderate, with a late onset. In 17 cases there were previous history of treatment with a ototoxic drugs, and most of them developed severe hearing loss. One of them was deaf-mute. No audometric differences between families with the homoplasmic and the heteroplasmic A1555G mutation were observed. CONCLUSIONS: Patients with the A1555G mutation generally present bilateral symmetrical sensorineural hearing loss, ranging from mild to moderate, slowly progressive, which is obvious approximately in the second decade of life and affects specially the high frequencies. Hearing loss severity is increased by treatment with aminoglycosides. PMID- 12584879 TI - [Olfaction disturbances in general ORL practice]. AB - Analysis of smell disturbances in the ENT outpatients department of in a General Hospital. A retrospective and descriptive study. We include all patients with olfactory alterations as the main reason for consultation. The sample was 38 patients. Viral, postraumatic, toxic, drug-induced and inflammatory. We performed anamnesis, nasal endoscopic and scan images on all patients. Olfactory disturbances were more frequent in women older than 55 (2:1). The viral cause was the most frequent aethiology (55.3%). Nasal endoscopic exploration was normal in 68.4% patients. The CT scan was the main imaging study used (68.4%). The younger patients recovered better than the older ones. Smell disorders provoked by virus, toxic and medical drugs do have a better. PMID- 12584880 TI - [Local sclerosing treatment with etoxiesclerol in ORL disease: Rendu-Osler disease, granuloma, angioma...]. AB - In a previous report, we showed the efficacy of the submucous sclerosant therapy with Etoxiesclerol (Polidocanol) in two patients with Rendu-Osler-Weber disease. We confirm, in this present study, the advantages of Etoxiesclerol in the treatment of the epistaxis in the Rendu-Osler disease, in six patients, 3 males and 3 females and likewise in two patients with non-specific granuloma of nasal septum and another patient with cavernous hemangioma of soft palate. PMID- 12584881 TI - [External lateral osteotomy in rhinoplasty for the correction of nasal dorsal deviations]. AB - We present a revision of 45 patients with nasal deviation operated on between 1993 and 2001. Septorhinoplasties were done in all cases using the external lateral nasal osteotomies technique. 67% of our patients were male and 33% were female with a mean age of 25. In all cases a septoplasty was performed. To correct nasal deviation, medial osteotomies through an intercartilaginous approach and external lateral nasal osteotomies were done. We did a postoperative follow-up of 35 patients. In terms of patient satisfaction, 78% of them felt that their nasal appearance had improved after surgery, and 22% felt that they had a great improvement. One case had to be reoperated on because of a traumatic nasal fracture after surgery. We believe this technique offers important advantages, such as: an excellent control of the fracture line, fewer incidences of open roof and lateral step, without causing visible scars. PMID- 12584882 TI - [Surgical tongue-base suspension for obstructive sleep apnea syndrome. Our experience]. AB - There are a lot of techniques involves to surgical treatment of the Obstructive Sleep Apnea Syndrome. Surgical treatments of tongue-base and hypopharyngeal colapse include glossectomy, hyoid suspension, mandibular osteotomy with genioglossal advancement and maxillay-mandibular advancement. Many or these procedures are associated with extreme morbidity. In this paper is evaluated the effectiveness of a minimally invasive technique for tongue-base suspension. The procedure was performed in 12 male patients whit O.S.A.S. that were evaluated with cefalometric and fiberoptic analysis. These preliminary results show the initial efficacy of this new surgical procedure. PMID- 12584883 TI - [Acute epiglottitis in adults. Our case load in 11 years]. AB - We report a retrospective study of 20 cases of acute epiglottitis in adults admitted at our hospital between 1991 and 2001. We gather a series of variables: Patient sex, age, year and month of admission, personal history, initial clinical symptoms, complementary tests asked, given treatment, evolution and hospital average stay. We found an obvious predominance in male (19:1 with respect to female), with an average age of 45 years. The main symptoms were dysphagia odynophagia (85%), followed by fever (55%) and pharyngocervical pain. Dyspnea was confirmed in 9 of 20 patients (45%); 5 of them required airway control with the help of intubation in one case, urgent coniotomy in three cases and tracheotomy in another one. A patient suffered from an acute mediastinitis and required assistance and medical i.v. extended treatment in UCI. We show a case of another patient who suffered sudden cardiorespiratory arrest a followed by death. He did not present previous dyspnea. Acute epiglottitis in adults is a rare pathology with a good prognosis in general, but may lead to an unpredictable and serious complication even in the absence of airway blockage. We carry out a medical literature review in this respect. PMID- 12584884 TI - [Intelligent system to perform a diagnostic protocol for lymphatic invasion in laryngeal cancer]. AB - Laryngeal carcinoma is the most frequent malignant tumour in head and neck. Node invasion is known to be one of the most important prognostic factors. The aim of this study has been to design an intelligent system to perform a diagnostic algorithm of metastasic neck nodes. 122 clinical reports of patients diagnosed of laryngeal carcinoma in our department have been reviewed. The compiled data have been: tumor site, T stage, N stage (clinical, after CT scan and post-surgery). The method used to design the intelligent system has been the ID3, which is able to generate a minimal decision tree. Palpation has been the variable that has given more information about node invasion. CT has proved to be more efficient in supraglottic tumours. ID3 method has shown to be useful in performing diagnostic algorithms, specially when the number of cases and diagnostic tests are high. PMID- 12584885 TI - [Subglottic MALT-type lymphoma: unusual location]. AB - We report a case of a 79-year-old woman presenting subglottic tumor whose hystopathological study was MALT-type lymphoma. After excluding the possibility of systemic dissemination, local radiotherapy treatment was applied. The interest of this study lies in the rare involvement of haematopoietic neoplasms in the larynx, especially the MALT-type in subglottic location. We review the literature on this subject, exposing the clinical, therapeutical and follow up data in respect to the initial location of the lymphoma. PMID- 12584886 TI - [Infrequent etiology of upper airway resistance syndrome]. AB - We present the case of a thirty-eight years-old man bearing of frequent crisis of asphyxiating wakefulness and breathnessless whenever he was asleep. There also were excessive daytime sleepiness and a strong snore before every wakefulness, but not for the rest of sleep. A polysomnographic study revealed normal values in the oxygen desaturation index, minimum oxygen saturation, an sleep structure alteration with a decrease of III-IV stages, high arousals index and RDI < 5. Pharyngeal diameters measured by mean of TC were normal, but sleepiness degree detected in the Epworth scale was high. Nasal fiberoptic endoscopy study allowed to see a swinging epiglottis that closed totally the upper airway during forced inspiration. The lack of desaturation episodes with numerous wakefulness along the sleep and excessive daytime sleepiness drove to the diagnosis of upper airway resistance syndrome (UARS). Treatment whit C-PAP just provoked an impairment in symptoms, but a CO2-laser partial epiglottectomy improved them. Patophysiological aspects of UARS, as well as its follow-up and therapeutic alternatives are commented. PMID- 12584887 TI - A short history of the development of vitreoretinal surgery. PMID- 12584888 TI - Prostacyclin and hormone levels in patients with symptoms of miscarriage and infection. AB - During pregnancy, acsending infection into the uterus, is followed by local increase of cyclooxygenase-2 (COX-2) activity, and consequently elevated prostaglandin production. Subclinical infections of fetal amniotic membranes are well known to cause preterm delivery. Spreading of infection from vagina and cervix into uterine cavity, can be tragic for the fetus. The aim of this study was to reveal wether lower genital tract infections are associated with symptoms of threatened miscarriage. Our investigation included 140 patients with symptoms of threatened miscarriage and 70 patients with uncomplicated pregnancies. Infections were detected by vaginal or cervical smears as well as tests for Chlamydial infection. Levels of prostacyclin, main prostaglandin product during pregnancy, were evaluated via its stable metabololite, 6-keto-PGF1-alpha. Both prostaglandin levels and hormones were determined by ELISA method. We found that serum values of prostacyclin were elevated, while levels of estradiol, progesterone and prolactin were significantly lower in patients with lower genital tract infections. Increased prostacyclin levels in pregnancies complicated by lower genital tract infections are caused by stimulation of COX/2 enzyme, due to elevated production of various cytokines which are possibly a compensatory mechanism resolving problems caused by bacterial endotoxines. PMID- 12584889 TI - [Olfactory perception in women with physiologically altered hormonal status (during pregnancy and menopause]. AB - INTRODUCTION: Olfaction is considered to be the ability to: perceive, conduct and recognize scents and odors. With its numerous connections to the limbic system and reticular formation, the olfactory system affects regulation of numerous vegetative functions, visceral functions and sexual behavior. Since estrogen and progesterone protect the olfactory function, changes in their levels in particular physiological states in women (in pregnancy and posmenopause) exert an influence on the ability to feel and recognize smells. It has its role in creating emotions and adjustment of visceral and vegetative response to particular emotional states. Also, it represents the connection between higher cortical functions and the endocrine system. MATERIAL AND METHODS: Our investigation was performed at the Ear, Nose and Throat Clinic in Novi Sad. The research included 80 healthy women classified into 4 groups; 20 women aged between 20 and 30; 20 women in the first trimester of pregnancy aged between 20 and 30; 20 premenopausal women aged between 41 and 50; and 20 women at least 3 years in postmenopause, aged between 41 and 50. For our research we used an olfactometer and the Fortunato-Niccolini method. RESULTS AND DISCUSSION: In pregnancy the thresholds of perception (TP) and identification (TI) of examined substances were slightly lower in comparison to nonpregnant women of the same ages, but without any statistical significance (p > 0.05). In climacteric postmenopausal women there was a significant decrease of olfactory ability in comparison to nonmenopausal women of the same ages (p < 0.01). CONCLUSION: All changes of the olfactory function in pregnancy are explained by mental changes of pregnant women as well as their hormonal status. Significant decrease of olfactory ability in postemnopause is explained by decline in sexual hormone levels. PMID- 12584890 TI - [Hyperhomocysteinemia--a risk factor for development of occlusive vascular diseases]. AB - INTRODUCTION: Occlusive vascular diseases take the first places on lists of diseases in general population today. In spite of this, all risk factors which contribute to development of these diseases are not yet known. Recent studies have shown that homocysteine plays a critical role in it and is established as a new risk factor. WHAT IS HOMOCYSTEINE?: Homocysteine is a sulfur containing amino acid formed in the metabolism of methionine. Reference values of homocysteine in circulation and different forms in plasma are described. HYPERHOMOCYSTEINEMIA--A RISK FACTOR: Homocysteine was associated with atherosclerosis and occlusive vascular disease in 1960s for the first time. Since then, many studies- prospective and retrospective, have confirmed the role of hyperhomocysteinemia as a risk factor in 42% of patients with cerebrovascular disease, 28% with peripheral vascular and 30% with coronary artery disease. The Physician's Health Study, a prospective study in which 15,000 male physicians took part, revealed that increase in homocysteine concentration of 1.7 mumol/l above normal values was associated by threefold higher risk for myocardial infarction. The risk for carotid artery stenosis also increases with elevation of homocysteine concentration. Hyperhomocysteinemia is associated with poor prognosis in patients with angiographically established coronary disease. Stroke, venous thromboembolism, and atherosclerosis in chronic renal failure are some of the complications of hyperhomocysteinemia. CAUSES OF HYPERHOMOCYSTEINEMIA: Hyperhomocysteinemia has numerous genetic and nongenetic etiologic factors. Cystathionine synthase deficiency, methylenetetrahydrofolate reductase deficiency and defects in the synthesis of cobalamin cofactors are genetically determined. Nutritional factors such as B12, folate or B6 vitamin deficiency, cofactors in homocysteine metabolism, lead to hyperhomocysteinemia. MECHANISMS OF HOMOCYSTEINE ACTION: Atherogenic propensity of homocysteine is related to endothelial dysfunction, blood thrombocyte aggregation changes in factors of coagulation. Oxidative stress is involved, but the exact mechanism is still unknown. CONCLUSION: Hyperhomocysteinemia is established as an important risk factor for occlusive vascular diseases. Reduction in homocysteine concentration can be achieved by supplementation of B-group vitamins, cofactors in homocysteione metabolism. Is it going to be effective in reducing cardiovascular risks remains to be seen. PMID- 12584891 TI - [Diagnostic importance of ultrasound in dermatology]. AB - INTRODUCTION: Utilization of 20 MHz ultrasound probes provided application of ultrasound in dermatology-dermatosonography. As a diagnostic tool, ultrasound was first registered in the early fifties of the past century. Great progress of dermatosonography occurred in the mid-nineties with introduction of the first 20 MHz scanner. METHODS OF ULTRASONOGRAPHY IN DERMATOLOGY: Several methods of ultrasonography have been developed: method A, method B, scannibg C method and Doppler ultrasound. They were based on different acoustic features of various tissues. ULTRASOUND IN DIAGNOSTICS: Today, dermatosonography is used in diagnosing: cutaneous neoplasms, inflammatory skin diseases and skin diseases of unknown etiology. Apart from that, it is used in differentiating benign and malignant tumors and may be helpful in monitoring morphea, dermatomyositis, cutaneous sarcoidosis, erythema nodosum, allergic diseases, postthrombotic syndrome, physiologic aging and aging due to sun exposure. It can be used in noninvasive monitoring of local treatment, therapeutic responses and adverse effects. CONCLUSION: Dermatosonography offers valuable possibilities in assessing skin morphology at an intermediate level: between clinical assessment and microscopy. PMID- 12584892 TI - [Treatment of acute upper respiratory tract infections in children]. AB - INTRODUCTION: Acute respiratory tract infections are the most common diseases of childhood. A preschool child suffers up to 5-7 infections of upper airways during a year. Upper airway infections make 80-90% of all respiratory infections. ETIOLOGY AND TREATMENT: In 75% of all cases respiratory infections are of viral etiology, 15% of bacterial and 10% are caused by mycoplasma, rickettsiae, fungi, parasites. The treatment of respiratory infections includes antimicrobial therapy (causal), relief of symptoms (symptomatic) and application of general principles of child treatment. The choice of antimicrobial drug is based on the evidence of agents and their sensitivity to antimicrobial drugs, age, patient's condition, previous treatment and possible allergic reactions to the drug. In cases where adequate specimen cannot be obtained for microbiologic tests, when these tests do not reveal the agent, or therapy must start before evidence of the agent is available, we must decide about the therapy, taking in consideration the most frequent agents, and those that would cause the most devastating clinical picture. This therapy can be modified later, according to the isolated agent and its sensitivity to the drug. Considering the incidence and importance of respiratory infections in morbidity and mortality of children, the aim of this article was to present guidelines in treatment of respiratory infections. The main point remains that the treatment should take into consideration the individual patient before all. PMID- 12584893 TI - [Psychological evaluation of patients with a nodular goiter before and after surgical treatment]. AB - INTRODUCTION: Association between endocrine and mental disorders has been recognized a long time ago, as well as their mutual dependence. The aim of this study was to evaluate the psychological structure of patients with thyroid nodule before and after surgical treatment. MATERIAL AND METHODS: In order to establish the type and degree of psychological disorders, we have examined 60 patients with thyroid nodule before and after surgical treatment by using DSM-IV classification of mental disorders and psychological instruments: semistructured psychiatric interview, MMPI, Zung Depression Scale and a list of panic symptoms. Patients with nonautonomous ("cold") nodules presented as euthyroid, and those with autonomous ("hot") nodules (after a period of drug therapy if they were hyperthyroid), required surgery. After a period of drug therapy two groups of patients were compared: group 1--euthyroid with "cold" nodules and group 2- euthyroid with "hot" nodules. Before surgical treatment both groups underwent psychological evaluation. Group 2 presented with: anxiety, depression and panic disorder with or without agoraphobia (72%, 46%, 28%) while group 1 presented with following results: 48%, 23%, 14% respectively. After surgical treatment patients from group 1 presented with psychologic disorders in less than 1%. DISCUSSION AND CONCLUSION: In some physical disorders, psychologic factors contribute directly or indirectly to the etiology, in others, psychologic symptoms are the direct results of a lesion affecting neural or endocrine organs. One of the key etiological factors is often a short-time or long-time stress and its direct consequence is altered functioning of various hormonal systems. Unexpected high percentage of psychological disorders in patients with thyroid disorders suggests that psychological evaluation before and after surgical treatment is unavoidable for good assessment and choice of treatment. PMID- 12584894 TI - Body fat mass in normal weight subjects. AB - Obesity is characterized by excessive body fat accumulation which may lead to serious health problems and complications. Body mass index is the most optimal parameter to evaluate the level of nutritional status and diagnose obesity. However, modern techniques studying body composition can more accurately determine whether the gain of body weight was on the account of body fat, lean body mass or total body water. If one's body mass index is in the range of normal values but the amount of body fat is above normal range, we talk about sarcopenic obesity. In order to evaluate presence of sarcopenic obesity, a group of 140 normal weight students of the Faculty of Medicine in Novi Sad were measured. Apart from standard anthropometrical parameters the amount of body fat was also determined by using the method of bioelectrical impedance analysis. Sarcopenic obesity was diagnosed in 25.71% of examined students. By using body mass index values this type of obesity cannot be diagnosed, and knowing that a higher amount of body fat in normal weight persons can lead to complications, especially metabolic, it is of great importance to evaluate the amount of body fat accurately. PMID- 12584895 TI - [Clinical characteristics and prognostic significance of preherpetic neuralgia]. AB - INTRODUCTION: Herpes zoster is a world-wide disease of older age commonly presenting with preherpetic pain. The aim of the study was to determine clinical characteristics of preherpetic neuralgia and its influence on occurrence of postherpetic neuralgia. MATERIAL AND METHODS: A prospective, controlled trial included 88 patients with preherpetic neuralgia. 44 herpes zoster patients without preherpetic neuralgia were included in the control group. All of them were clinically followed-up for three months after complete healing of skin lesions. RESULTS: Older age (> 60 years) was significantly predominant (59.1%) compared with other age groups (p < 0.01) as well as female sex (59.9%) compared with the male sex (p < 0.01). There was no significant predominance of any type of preherpetic neuralgia (stabbing, burning, itching, dull pain). More intense preherpetic pain (reported as "severe" and "moderate") was established more often than mild pain. The mean duration of preherpetic pain was 4.4 days (ranged between 1-20 days). Postherpetic neuralgia developed in 36/88 patients with preherpetic neuralgia (affecting predominantly older than 50 years of age- 31/36), but there was no significant difference in proportion of postherpetic neuralgia (PHN) according to those without preherpetic neuralgia. DISCUSSION AND CONCLUSION: People older than 60 years are the most common age group among herpes zoster patients suffering from preherpetic neuralgia. Sex distribution of patients with preherpetic pain reveals highly significant predominance of female sex. Opposite to some other authors' reports, preherpetic neuralgia and its severity have not been proven as risk factors for postherpetic neuralgia in patients involved in our trial. PMID- 12584896 TI - [Endometriosis and pain]. AB - INTRODUCTION: The main symptoms of endometriosis are pain, adnexal tumor and infertility. Pelvic pain and dysmenorrhea are cardinal symptoms as well as pain upon defecation, suprapubic pain (dysuria), pain during coitus and during gynecologic examination. Pain can be caused by fibrotic reaction of the adjacent tissue, adhesions, prostaglandins produced both in endometrium inside the uterus and ectopic endometrium, and also by increased macrophages in the peritoneal fluid. MATERIAL AND METHODS: 500 infertile patients underwent the procedure of laparoscopy in order to determine the cause of infertility. The control group comprised 200 fertile women. The presence of pain in the small pelvis was compared in two groups of patients: with or without endometriosis. Correlation of pain with the stage of disease and location of endometriotic implants in the small pelvis has also been investigated. RESULTS: Endometriosis was diagnosed by laparoscopy in 26% of infertile and 5% of fertile women. The difference was statistically significant (p < 0.001). Dysmenorrhea was present in 46.92% of infertile women with endometriosis and in 48.68% of women without endometriosis. The difference was not statistically significant. Dysmenorrhea was present in 30% of fertile women and in realtion to infertile women, the difference was statistically significant (p < 0.05), no matter if they have endometriosis or not. In relation to stages of endometriosis (the revized classification of the American Fertility Society), there is no significant difference concerning dysmenorrhea. Dysmenorrhea occurred in the first stage in 48%, in the second stage in 44.19%, in the third stage in 50% and in the fourth stage in 44.44%. The correlation coefficient ranged from -8.85 to -0.89. The correlation existed, it was high, but negative. Sensitivity, specificity and prognostic value of symptoms of dysmenorrhea have been estimated in relation to endometriosis with following results: sensitivity was 47%, specificity 51% and the prognostic value i.e. the possibility of occurrence in patients with dysmenorrhea was 25%. Localization of endometriotic foci did not affect occurrence of pain symptoms. PMID- 12584897 TI - [Circadian rhythm of blood pressure and obesity--blood pressure variation and obesity]. AB - INTRODUCTION: The association between obesity and arterial hypertension has been established in a great number of studies. Our objective was to investigate whether circadian rhythm of blood pressure is disturbed among obese people. MATERIAL AND METHODS: In this cros-sectional, randomized study, Schiller BR-102 device was used for ambulatory blood pressure monitoring. One hundred and twenty outpatients were divided into three randomized groups: obese body mass index 30 kg/m2 (52 patients), overweight (28 patients), with body mass index 25.0-29.9 kg/m2 and normal weight (control group) (48 patients), with body mass index 18.5 24.9 kg/m2. In all patients we investigated the following blood pressure parameters: average blood pressure (total, day-time and night-time), maximal blood pressure and dipping or non-dipping blood pressure pattern during night (for systolic and diastolic blood pressure, respectively). RESULTS: In body mass index beyond 30 kg/m2 only systolic blood pressure parameters were significantly higher--average blood pressure--during daytime (P = 0.034) and during night (P = 0.014); maximal blood pressure (P = 0.001). In body mass index beyond 30 kg/m2, absence of normal blood pressure during night was significantly more often registered (P = 0.007). DISCUSSION AND CONCLUSION: The non-dipping blood pressure pattern and increase of systolic blood pressure only reveal hyperactivation of sympathetic nervous system as a leading patophysiological mechanism causing arterial hypertension in obese patients. PMID- 12584898 TI - [Gastrointestinal stromal tumor: case report]. AB - INTRODUCTION: Gastrointestinal stromal tumors represent extremely rare tumors of the gastrointestinal system, especially when localized on the small intestine. CASE REPORT: We report a case of a female patient, with recurrent gastrointestinal bleeding and severe anemia, caused by gastrointestinal stromal tumor of the small intestine. After negative endoscopic findings, she underwent radiological examination of the small intestine. Primary diagnostic radiological evaluation included: small intestine passage enteroclysis, computed tomography of the abdominal cavity and selective angiographic study of the three major aortic branches that supply the gastrointestinal tract in the abdomen (celiac axis, superior mesenteric artery and inferior mesenteric artery). Secondly, ultrasound of abdominal cavity was performed. Findings of small intestine passage and enteroclysis were negative. The tumor was visualized by computed tomography and ultrasound, but without distinctive anatomical localization in the abdominal cavity. DISCUSSION: The diagnostic dilemma has been resolved by using selective angiographic examination of celiac axis and superior mesenteric artery and thus a tumor formation was visualized in the mesenterium of the small intestine. Radiological findings were confirmed by surgery. Histopathological findings were positive for gastrointestinal stromal tumor. CONCLUSION: Gastrointestinal stromal tumors of the small intestine rarely cause recurrent bleeding, but they should be included in differential diagnosis. PMID- 12584899 TI - [Importance of early rehabilitation of a child with severe multiple injuries: case report]. AB - CASE PRESENTATION: This is a case presentation of an eight-year-old boy, a traffic accident victim with severe polytrauma. The boy was admitted to hospital with lacerations and contusion injuries of the left basal frontal and left temporal brain region, moderate brain tissue edema and fracture of the left clavicle which were diagnosed by CT (computer tomography). The boy was in coma (Glasgow Coma Scale--5). Oral feeding and catheter removal were established after one month. Three months later his state of consciousness improved, he could carry out simple orders, started vocalization and visited ambulance by wheel chair. Speech and independent walking were achieved 4 months after the accident. PHYSICAL THERAPY: Physical therapy included: neural-stimulation by Vojta and sclerodermal message from seventh day. Later on, speech and work therapy were applied. DRUG THERAPY: Drug therapy included: muscle relaxants and tranquilizers while orthopedic treatment meant plastic immobilization. Control CT revealed chronic subdural hemathoma in the right frontal-parietal region. The boy could walk, talk, have normal diet, and controlled sphincters four months after severe trauma. Now, he is functional and on the level of moderate mental retardation. Early rehabilitation was extremely important in his recovery. PMID- 12584900 TI - [Child health care and its development in Vrsac]. AB - Documents concerning history of medicine during the Turkish reign (1552-1716) are very rare. However, there is evidence of plague epidemic in 18th century and colera epidemic in the 19th century. The first medical institutions: The German Communal Hospital, The Serbian Hospital and the Pharmacy were founded in the second half of the 18th century. In the year 1803, children were vaccinated against variola. The first Serbian book about child care--"Cadoljub" was written by Dr. Gavrilo Pekarovic (1812-1851) during his studies of medicine in Budapest. In 1927 the city founded a dispensary for the newborn. The Polyclinic for schoolchildren was established as a part of the Health Center in 1934. After World War II, Children's Department was opened in the Health Center, later on it was turned into Mother and Child Center. At the beginning of 1955, a provisional children's ward with 18 beds was established in the former sanatorium, whereas till the end of the year it had 49 beds. In May 1965, it was moved into a new hospital building. After integration of Hospital and the Health Center into a Medical Center in 1967, a department for children was founded and it consisted of the emergency center and a hospital. Parents counseling, dispensary for children and dispensary for schoolchildren were founded in August 1971. PMID- 12584902 TI - [Erasme is 25 years old]. AB - The opportunity of the 25th anniversary of the Erasme hospital was taken for drawing up an inventory of the scientific publications issued from the medical teams of the institution. The inventory was established in accordance with the missions of an academic hospital, namely, evaluation and development of new diagnostic and therapeutic tools, creation of new knowledge in basic science as well as in clinical research, translation of new biomedical findings to patient's care by the fruitful back and forth between the laboratory and the bedside and, finally, launching of new skills to be prepared for the medicine of the new millennium. The assessment is positive. However, the future is uncertain due to the under reimbursement of the academic medicine. PMID- 12584901 TI - [Principles of management of high-energy injuries of the leg]. AB - INTRODUCTION: High-energy traumas are open or closed injuries caused by force (missile, traffic injuries, crush or blust injuries, falling from heights), affecting the body surface and transferring high amount of kinetic energy inducing great damage to the tissue. Management of such lower extremity injuries has evolved over past several decades, but still remains a difficult task for every surgical team. Specific anatomic and functional characteristics combined with extensive injuries demands specific treatment protocols. MULTIPLE INJURIES: In a multiple injured patient the first priority is management of life threatening trauma. Despite other injuries, surgical treatment of limb threatening injuries must start as soon as life-threatening condition has been managed. TREATMENT ALGORITHMS: Algorithms are especially beneficial in management of severely injured, but salvageable extremities and in making decision on amputation. Insight into mechanisms of injury, as well as systematic examination of the affected limb, should help us understand the extensiveness of trauma and make an adequate management plan. PREVENTION OF INFECTION AND SURGICAL APPROACH: Prevention of wound infection and surgical approach to high-energy limb trauma, which includes wound extension, wound excision, skeletal stabilization and if necessary muscle compartment release, should be done in the first 6 hours after injury. METHODS OF SOFT TISSUE RECONSTRUCTION: Commonly used methods for soft tissue defects must provide wound coverage in less than five days following injury. REHABILITATION: Early passive and active mobilization and verticalization of patients is very important for successful treatment. CONCLUSION: Good and timely evaluation of the injured and collaboration between plastic and orthopaedic surgeons from the beginning of treatment, are crucial for final outcome. PMID- 12584903 TI - [The pathology department]. AB - The evolution of the Laboratory of Pathology at Erasmus Hospital is directly related to the morphological diagnosis integration in the medico-surgical organization of the Hospital. Such integration is based on the creation of links between the Laboratory of Pathology and the clinical departments and necessitates permanent adaptations to new complementary technology. Immunohistochemical methods were gradually included in the surgical morphological diagnosis. Currently, new markers from molecular biology are needed to obtain accurate pathological diagnosis. We observe an increasing hyperspecialization of the morphological classification with complete integration of such biological markers leading to the necessity of carrying out collegial diagnosis. The development of the telepathology technology permits international collegial diagnosis. According to its central position the Laboratory of Pathology plays an important part in the development of the clinical and fundamental research. The research subjects of the Laboratory of Pathology concern diagnostic and prognostic evaluation in colic cancers, sarcomas, brain tumors and HPV related tumors endometriosis. PMID- 12584904 TI - [The anesthesiology-recovery department]. AB - The Department of Anesthesiology and Reanimation is organised in units with clinical activities, which include the pre-operative care of patients, anesthesiological care and immediate post-operative supervision. Two post operative treatment rooms also form part of the department. The main fields of research of the various units result from collaborations with other departments of Hopital Erasme, in particular with regard to the development of advanced techniques or fit within the confines of the speciality. PMID- 12584906 TI - [The medical chemistry department]. AB - The laboratory of clinical chemistry performs more than 300 different tests in biochemistry, hormone and tumor markers analysis, therapeutic drug monitoring and toxicology. For the most basic tests it has followed the trend of clinical chemistry towards automation and since 2001 the heart of the laboratory is a modular automated system (MODULAR) including a preanalytical platform, unique in Belgium. For more sophisticated tests, the most recent techniques have been implemented, in particular capillary electrophoresis and ICP-MS ("inductively coupled plasma-mass spectrometry). Since 1994, the laboratory has become a reference center in the field of erythrocyte hereditary diseases, combining screening, diagnosis and research. The other research themes are the physiopathology of first trimester pregnancy and the P2Y receptors of extracellular nucleotides. PMID- 12584905 TI - [The cardiology department]. AB - Cardiology was present since the very beginning of the Erasme hospital. The Department of Cardiology was created in 1979. This department is made of a hospitalization unit (54 beds), a coronary care unit (12 beds) and a large technical unit. Clinical activity has increased tremendously and this had led to an important research activity in the fields of cardio-respiratory and metabolic adaptation to exercise, particularly after cardiac transplantation, of cardiovascular epidemiology, of clinical pharmacology and in pulmonary as well as systemic hypertension. PMID- 12584907 TI - [The heart surgery department]. AB - The initial development of cardiac surgery at Erasme Hospital was closely related to the achievements in thoracic organ transplantation, with numerous synergies between other clinical and research units of the Faculty of Medicine. New advances in biology and biotechnology have met the challenges of modern cardiology, in the fields of advanced heart failure, refractory angina, rhythm disturbances or minimally invasive surgery. Fundamental aspects of clinical practice have been the subject of laboratory investigations, resulting in fruitful interactions and promising scientific outlooks. PMID- 12584908 TI - [The plastic surgery department]. AB - Plastic surgery associates reconstructive and aesthetic surgery, as these are complementary and cannot be dissociated. Modern techniques in reconstructive surgery include grafts and flap transfers. Our team does 4500 consultations and 750 operations per year, out of which a large proportion of complex reconstructions. In order to treat very various diseases, strong collaborations exist with several units such as orthopaedics, ENT, maxillofacial surgery, neurosurgery, ophthal-mology, gynaecology, and dermatology. PMID- 12584909 TI - [The thoracic surgery department]. AB - The Department of Thoracic Surgery has been developed since 40 years, and particularly since 23 years at the Hopital Erasme. The high rate of lung cancers did induce an important activity in surgical oncology, inside multidisciplinary teams. After a first attempt in 1969, lung transplantation is now successful. Infectious diseases, traumas and functional respiratory defects impose heavy daily charge. Research has been developed in lung transplantation, thyroid metabolism, respiratory mechanics and clinical trial in oncology. PMID- 12584910 TI - [The dermatology department]. AB - The department of dermatology has developed a broad clinical activity with a particular expertise in oncology, allergology, psoriasis and gynecological dermatology. Research has also been developed in these areas. PMID- 12584911 TI - [The endocrinology department]. AB - The clinical activities of the department of endocrinology encompass the care and treatment of diabetes, thyroid diseases, hypothalamo-pituitary, adrenal, gonadic and parathyroid diseases, obesity, hypercholesterolemia and paraneoplastic endocrine syndromes. These domains are briefly described. The research activities of the department have investigated the regulation of thyroid metabolism in vitro, the intrathyroid H2O2 generating system, the physiopathology of toxic thyroid nodules and the effects of ageing on the thyreotropic function of the normal ageing male. Studies of "jet lag" conditions have shed a new light on hormonal chronophysiology. Other investigations have considered the regulation of ketone body metabolism, the relationship between nutritional status and glucose metabolism, and some aspects of immunodiabetology. PMID- 12584912 TI - [The hospital epidemiology and hygiene unit]. AB - The hospital Epidemiology and Infection Control Unit aims at preventing nosocomial infections and controlling epidemics. In this field, several researches have contributed to improve the knowledge of infections secondary to biliary endoscopy, of epidemics of nosocomial Legionella and multiresistant gram negative and Staphylococcus aureus infections. Studies about the rational use of antibiotics have contributed to a better control of the transmission of multiresistant pathogens. PMID- 12584913 TI - [The surgical gastroenterology department]. AB - The Department of Digestive Surgery was born in 1977. It is a part of the medical surgical unit of gastroenterology and hepatopancreatology. The various developed sectors concern hepatic surgery and liver transplantation (treatment of hepatic tumors and cirrhosis), pancreatic surgery and surgery of the biliary tract (treatment of benign and malignant pancreatic tumors, tumor of the biliary tract, chronic pancreatitis and biliary stones), surgery of morbid obesity (gastroplasty or gastric by-pass), surgery of the upper digestive tract (benign and malignant tumors of the oesophagus or the stomach, treatment of gastroesophageal reflux), surgery of the abdominal wall, colorectal surgery and surgery of the inflammatory bowel diseases (colorectal cancer, familial polyposis, Crohn's disease, ulcerative colitis), proctologic surgery and surgery of anorectal functional disorders, neonatal and paediatric surgery. PMID- 12584914 TI - [The medical gastroenterology department]. AB - The present paper summarizes the various themes of research which have been developed in the department of medical gastroenterology since it was created in 1977. These include: in pancreatology, the study of chronic pancreatitis pathogenesis, acute pancreatitis pathogenesis and immunomodulation, endoscopic treatment of chronic pancreatitis, the development of new imaging techniques of the bile ducts and the pancreas, as well as the treatment of pancreatic cancer and benign or malignant biliary diseases. in hepatology, the immunomodulation of liver cirrhosis, especially alcoholic liver disease, the modulation of experimental acute and chronic hepatitis, the study of liver ischemia reperfusion. Clinical hepatology has focused on liver transplantation, prognosis factors of chronic liver disease and treatment of portal hypertension and viral hepatitis. in gut diseases, the treatment of gastro-oesophageal reflux and its complications, the therapeutic endoscopy of the upper and lower GI and the prevention, as well as the treatment, of colon cancer, the pathogenesis and the immunopharmacology of inflammatory bowel diseases and the clinical enteral and parenteral nutrition. PMID- 12584915 TI - [Medical genetics service]. AB - Created in 1987, the department of medical genetics finds its origins in molecular endocrinology research which had developed from the seventies at the Institut de Recherche Interdisciplinaire en Biologie Humaine et Moleculaire (IRIBHM) of the Faculty of Medicine. After its fusion with the Center of Human Genetics of the ULB, in 1992, the department is composed of three units: the lab of molecular genetics and oncology, the lab of cytogenetics and a clinical genetics unit. One thousand consultations of genetic counseling and more than 15,000 molecular or cytogenetic diagnostic procedures are performed annually. The development of the clinical activities was paralleled by a very active research activity, resulting in a series of "firsts". Amongst the main results are: the identification of the first mutations responsible for congenital hypothyroidism; the molecular cloning of the TSH receptor and of a series of "orphan" G protein coupled receptors; the identification of a novel neuropeptide, nociceptin, by the first example of "reverse pharmacology"; the identification of olfactory receptors on the sperm of mammals, including man; the identification in molecular terms of the mechanisms responsible for acquired and hereditary hyperthyroidisms; the identification of the chemokine receptor CCR5 as the major coreceptor of HIV 1, and of the prevalent mutation of CCR5 conferring resistance to HIV to about 1% of the European population. PMID- 12584916 TI - [The gynecology-obstetrics department]. AB - The scientific and clinical activities of the Department of Obstetrics and Gynaecology have involved the three main subdivisions: the gynecological surgery, the obstetrics and fetal medicine, the endocrinology and the reproductive medicine. Minimal invasive surgery including laser assisted laparoscopy or robotic assisted surgery has been particularly developed. Endometriosis, a frequent and sometimes particularly invasive disease, and oncologic surgeries have been developed in collaboration with the digestive surgery department. The department has also contributed to the comprehension and treatment of prenatal pathologies such as premature labor and deliveries or the gestational diabetes. The department has supported the development of techniques to study the fetal well-being in utero: the prenatal echography, the chorionic villous sampling, the amniotic puncture or the cordocentesis for prenatal genetic diagnosis or fetal infectious contaminations, the CMV transmission more specifically. In endocrinology and reproductive medicine, the department has mainly developed the in vitro fertilization techniques. The prolonged embryo culture, the study of preimplantation embryo metabolism, the preimplantation genetic diagnosis and the cryopreservation of ovarian fragments to preserve fertility in women undergoing oncologic treatments represent the more recent developed topics. Finally, the security of viral transmission in assisted procreation and the treatment of these patients with chronic viral diseases (Hepatitis C or HIV) are another domain with important scientific activity. PMID- 12584917 TI - [The hematology clinic]. AB - Clinic of Haematology, opened in 1993 is now performing each year about 30 blood stem cell transplantations, managing more than 8,000 hospital days and about 2,300 consultations. We are involved effectively in the EORTC Leukaemia Group, the European Bone Marrow Transplant Group (EBMT), the IFM and the GELA interactive groups. Major scientific contributions interested the management of peripheral blood stem cell transplantations, the study of multidrug resistance (MDR) in hematologic malignancies, the treatment of lymphoproliferative diseases by monoclonal antibodies, purification of autotransplants by positive selection in multiple myeloma and the expansion of new ways of administration of purine analogs in chronic lymphocytic leukaemia. PMID- 12584918 TI - [The medical imaging and radiology department]. AB - Technological developments arising from research have affected the whole wide spectrum of medical endeavor and have made a very significant impact on clinical practice and especially on imaging sciences. Ultrasonography brought spectacular advances, but CT and MRI became important landmark techniques. A further important development, which greatly increased the involvement of radiologists in direct patient management, was the growth of interventional and therapeutic techniques, called interventional radiology. Some statistics: approximately 155,000 patients per year including 19,000 CT Scans, 10,000 MRI exams, 21,000 ultrasound examinations and 1,000 therapeutic procedures. Some research activities: CT quantification of pulmonary emphysema, respiratory mechanics, MR and CT angiography, antenatal diagnosis of congenital and genetic diseases of the fetus, quantification of portal haemodynamics, MR imaging of bile and pancreatic ducts, morphologic and functional imaging of the brain, radiology of bone trauma, MR characterization in hepatic lesions. PMID- 12584919 TI - [The immunodeficiency treatment unit]. AB - Physicians of the unit have first taken care of patients with acquired Immunodeficiency in 1981. We have become an independent "Reference Centre" in 1998. The multidisciplinary team follows more than 300 patients on a regular basis. AIDS has been amply publicized, but other immuno-deficiencies have not. Primary immunodeficiencies are "orphan" diseases; they can be as serious, or more severe even, than AIDS. About 60 patients with "PID" are followed by the team. We are involved in research, and have participated in the identification of a mutation of an HIV co receptor that protects against HIV infection. We also studied the pathogenesis of Kaposi's sarcoma, and the immunological basis of adverse reactions to intravenous gammaglobulins. PMID- 12584920 TI - [The immunology-hematology-transfusion department]. AB - New immunotherapies derived from biotechnology offer fascinating perspectives in different fields of medicine including anti-infectious vaccines, cancer, organ transplantation and autoimmune diseases. In this paper, we illustrate how the Department of Immunology can contribute to the development of these new treatments within a academic hospital such as the Erasme Hospital at the Universite Libre de Bruxelles. PMID- 12584921 TI - [The infective diseases clinic]. AB - The Clinic of Infectious Diseases was created at Erasme Hospital in 1980. Its main activity consists in taking care of patients with infectious complications in various hospital departments. Its main scientific contributions concern the evaluation of new treatments, pharmacokinetics of antimicrobial agents and physiopathology of sepsis. The evaluation of the impact of infectious diseases on the appropriateness of antimicrobial therapy and the control of infection is a major contribution of the clinic. Several works concern the study of the immune response in case of infection due to an intracellular pathogen and in response to vaccination. PMID- 12584922 TI - [The general internal medicine department]. AB - The Department of General Internal Medicine is devoted to the evaluation of patients with autoimmune systemic diseases, multiorganic disorders or presenting non specific symptoms such as chronic fatigue, unexplained weight loss or fever of unknown origin. The interest in salt and water metabolisms had led to original contributions in the treatment of hyponatremia in man and to the understanding of the osmotic demyelinating syndrome in a rat model of hyponatremia. The study of ageing in man and rodents had contributed to better understand lymphocyte and thyroid function in the elderly. The care for patients with various autoimmune disorders led to original observations in the pathogenicity of Sjogren's syndrome adult onset Still disease or sarcoidosis as well as the follow up of patients treated with azathioprine. Intensive collaboration with the Department of Immunology led to identify Th2 clonal lymphocytes as the cause of the so-called idiopathic hypereosinophilic syndrome in some patients and to define the clinical and biological features in this subset of patients. PMID- 12584923 TI - [The nuclear medicine department and the TEP/Biomedical Cyclotron unit]. AB - During the last 25 years, the clinical and experimental activity in nuclear medicine at Erasme hospital has been influenced by the implementation of positron emission tomography (PET) in 1990 as a method of brain functional investigation. The activity of the PET/biomedical cyclotron unit has been dedicated to various subjects in neurology, neurosciences, psychiatry, oncology and cardiology. This has been made possible by developments in radiochemistry. The radiochemistry laboratory has designed and produced original tracers such as 9-[(3-[18F]fluoro-1 hydroxy-2-propoxy)-methyl]guanine (FHPG), a tracer of viral thymidine kinase activity in gene therapy protocols. We have brought new applications of PET, such as its integration into stereotactic neurosurgical and radioneurosurgical techniques in order to improve their diagnostic and therapeutic performance in neurooncology. We have also conducted multiple studies on brain physiology and pathophysiology, in particular with the use of functional and metabolic brain mapping methods and the use of tracers of neurotransmission systems. The Department of nuclear medicine has also performed studies on bone metabolism and investigated in vivo imaging methods of infectious and immune processes. PMID- 12584924 TI - [The microbiology department]. AB - From the outset, the Department of Microbiology of Erasme Hospital has striven to offer to clinical teams a full range of high performance tests for diagnosis and therapeutic monitoring of bacterial, viral, fungal and parasitic infections, including opportunistic infections. Furthermore, the laboratory has developed and made available cutting edge tools for the epidemiologic surveillance and outbreak investigation in support to the hospital infection control and antibiotic resistance control programmes. Research and development programmes have led to technological innovation in the fields of molecular virologic diagnosis, notably for HIV and herpes viruses, rapid bacterial identification, detection of resistance genes and epidemiological typing. Research in collaboration with clinical teams has focused on congenital infections, HIV infection, opportunistic infections and prevention of nosocomial infection. The Department acts as national reference laboratory for a number of infectious diseases and co ordinates or actively participates to several European multicentre studies and epidemiologic surveillance networks. PMID- 12584925 TI - [The intensive and non-intensive neonatal care department]. AB - To be born very prematurely in 2002 is very different of to be born very prematurely in 1978: the progress of the fetal and perinatal care have, amongst others, decrease the mortality of the neonates with a birthweight below < 1,000 g from 61% to 12%. The technological progresses in artificial ventilation have led to a significant decrease in chronic lung disease down to 5% or less, and of cerebral complications (intraventricular hemorrhages, grade 3 and 4 and/or periventricular leucomalacia) down to 11% or less. The progress in surgery and anesthesiology have allowed us to operate the extremely low birthweight infants in the neonatal unit when needed. This has been possible thanks to a multidisciplinary team approach: many specialists working together from conception to birth and from birth to home try to offer the best to these sometimes very small human beings. PMID- 12584926 TI - [The medico-surgical department of nephrology dialysis and renal transplantation]. AB - The Department of Nephrology of the Hospital Erasme, opened 25 years ago, is now performing, each year, 22,000 hemodialysis sessions, 800 patient-weeks of peritoneal dialysis treatment and 70 renal grafts. Scientific contributions of the department deal with vascular access for hemodialysis, susceptibility to infections of dialyzed patients, parathyroid surgery, biocompatibility of dialysis membranes, predictive factors of renal graft survival, immunosuppression with monoclonal antibodies, experimental studies of graft tolerance and rejection, toxic nephropathies. The most original contributions are related to anaphylactoid reactions in hemodialysis by association of acrylonitrile membranes with inhibition of the converting enzyme, to advantages and side effects of OKT3 monoclonal antibody and to discovery and study of the Chinese herbs nephropathy. PMID- 12584927 TI - [The neurosurgery department]. AB - Opened on November 1st, 2001, the Department of Neurosurgery has progressively grown to become worldwide renown in a few years. All the pathologies are covered, from lumbar disc hernia to intracranial tumors and vascular malformations. But the originality stays into the exceptional environment by the concentration of logistic resources and the ability of clinician and researchers who daily collaborate with the neurosurgical team. The Department of Neurosurgery has a strong reputation in several fields like intraspinal cord tumors or Pet-guided Neurosurgery in stereotactic biopsies, neuronavigation and Gamma Knife and, generally speaking, in the original approach of the treatment and follow-up of brain tumors. Neurodegenerative diseases also benefit of modern approaches trough the Gamma Knife, deep brain stimulation or fetal cell grafting into the brain in Parkinson and soon in Huntington diseases. Last but not least, the arrival for the 25th anniversary of Erasme Hospital of an interventional MRI will allow to follow in real-time the resection of brain tumors with an obvious benefit for the surgical performances and the quality of life of the patients. It will also open a new window for neurosurgical research through combination with functional MRI and Pet-Scan, reinforcing the reputation of Erasme Neurosurgical Department who has been distinguished in 1997 by the World Health Organisation as "WHO Collaborating Center for Research and Training in Neurosurgery" and nominated again in 2002 for a new 4-year period, which is unique in the Neurosurgical World. PMID- 12584928 TI - [The neurology department]. AB - The department of neurology is devoted to the diagnosis and medical treatment of organic diseases of central nervous system (brain and spinal cord) and peripheral nervous system (peripheral nerves and muscles). Basic and clinical research in neuroscience constitute an essential activity of the department that defines its academic character. Over the years, the department of neurology has evolved from providing general neurology services to a multifaceted unit that has developed the several subspecialties of clinical neuroscience. Main research areas have included neurooncology, neurophysiology, neuropsychology, cerebrovascular diseases, childhood epilepsy and conditions affecting the psychomotor development of children. Neurogenetics is a recent addition to the areas of the interest of the department; research in neurogenetics includes basic investigations as well as clinical studies and focuses on inherited ataxias and genetic epilepsies. PMID- 12584929 TI - [The medical oncology clinic]. AB - The clinic of medical oncology is mainly devoted to the development of new anticancer treatments based on molecular biology and immunology. The clinic was the first in Belgium to start a protocol of gene therapy. Scientific contributions deal with the role of various oncogens in cell transformation, the interaction between cancer and the immune system and, new tools for the molecular diagnosis of cancers. Focus was particularly put on the development of new vectors for gene therapy and antitumor cell vaccines for cell therapy. PMID- 12584930 TI - [The ophthalmology department]. AB - The main clinical activities developed during the first 25 year of the department are described. Topics of fundamental and also clinical research are briefly reviewed. PMID- 12584931 TI - [The oto-rhino-laryngology and cervico-facial surgery department]. AB - ENT department was present at the opening of the hospital but its activity began to increase in 1989. Now all the fields of modern ENT are present due to the collaboration of physicians very specialized in otology, rhinology, laryngology and head and neck surgery. Drilled in all surgical techniques using the best technologies available, the members of our team are also working in collaboration with the Laboratory of Histopathology in the field of nasal inflammation, cholesteatoma and malignant tumors of head and neck area. PMID- 12584932 TI - [The orthopedics-traumatology department]. AB - The Department is involved in clinical and research activities. We pioneered the clinical application of "smart of Orthopaedic implants". The monitoring concept, bringing a high added value, helps in assessing fracture healing or implant fixation. The technique, featuring the 21st century, also requires advanced knowledge in biomaterials. Such implants have been used during the ESA Parabolic Flight Campaigns. These measurements are a prerequisite to establish prevention programs of the bone demineralisation during space flights but also in disuse osteoporosis. The Department is deeply involved in external fixation. In vitro studies of the mechanical insertion and holding parameters of external fixation pins have been performed to produce new implants, which efficiency has been proved by a clinical study. We defined original clinical principles of external fixation as its use in upper limb indications, to maintain microsurgery transplant, preserving devascularization and providing adequate fixation. Our biomechanical approach of the upper limb, now confirmed by others research groups, questioned old clinical concepts. The isokinetic evaluation assesses the dynamic function of articulations, i.e. after arthroplasty, and is also used for medico-legal evaluation. The clinical research touches orthopaedic (or systemic) diseases as algodystrophy, Kienbock disease, Kashing Beck. We conducted a multidisciplinary research on the effect of electromagnetic fields (EMF) on cells differentiation. In addition to possible healing properties, it helps as an objective support to analyse the exposure to the environmental EMF. PMID- 12584933 TI - [The medico-surgical department of vascular disease]. AB - The Department of Vascular Diseases can be defined as a multidisciplinary integrated clinical entity involving internists and surgeons interested in patient care, education and research in the field of arterial, venous and lymphatic diseases. Because of the increasing medical complexity with the proliferation of treatment options available, the combined expertise of specialists from different training background is required to provide the optimal patient management. The truly integrated entity with interactions on daily basis between internists and surgeons also provides opportunities for improvement in the training of fellows in vascular medicine. Basic and clinical research is focused on thrombosis and atherosclerosis. The main topics that have been developed include: the pathophysiology of thrombosis, atherogenesis and the diagnosis and treatment of venous thromboembolic disease. The successful development of our Department demonstrates that the combined expertise of internists and surgeons has resulted in marked improvement in the efficiency of patient management. PMID- 12584935 TI - [The psychiatry department]. AB - The Department of Psychiatry was first opened in April 1978. It is one of the largest sector of the hospital and contains 90 beds including a sleep laboratory and an adolescent unit. The clinical activities are broad and multidisciplinary, including novel psychopharmacological and psychotherapeutic and psychosocial approaches. Educational and research activities have also been developed, in particular in the areas of behavioral genetics, sleep physiology, brain imaging and psychosocial research and training. PMID- 12584934 TI - [The pneumology department]. AB - The department of pneumology of the Erasme hospital exists since 25 years. The basic clinical activities include pulmonary function testing (7,500 patients per year), endoscopy, including interventional endoscopy (1,500 patients per year), thoracic oncology, allergology, rehabilitation and aid to smoking cessation. The following expertise fields have been largely developed: lung transplantation, treatment of cystic fibrosis in collaboration with the children's hospital Reine Fabiola, occupational. PMID- 12584936 TI - [The rheumatology and physical medicine department]. AB - Actually, 18 rheumatologists and specialists in physical therapy are collaborating in the Department, allowing to develop possibilities for the diagnosis and therapeutic challenge of patients suffering from disorders of the locomotor system. Accordingly, the knowledge, the know-how, the experience plus the willingness to make a good job are used for helping the patient, for contributing to medical progress and also for the education of future medical doctors. Our department has significantly contributed to a better understanding and therapeutic approach of rheumatoid arthritis, Sjogren's syndrome, aseptic necrosis, osteoporosis, osteoarthritis, and inflammation; it has been among the first in the world to offer new therapeutic modalities, otherwise not accessible and, to help a series of hopeless patients. In addition, a new sector for a performing rehabilitation has been recently developed. Accordingly, during these mast twenty years, a performing department with a motivated team has been developed offering a maximum of medical services for the community and ready for the challenges of tomorrow. PMID- 12584937 TI - [The intensive care department]. AB - Intensive Care Departments are designed for the care of the critically ill, and are equipped with all the material necessary for monitoring and treatment, as well as specially trained medical, nursing, and paramedical staff. With a team of more than 170 highly qualified staff, the Department of Intensive Care at Erasme Hospital, has acquired a national and international reputation in the fields of clinical and experimental research and education, as well as for its standards of clinical practice. Clinical and experimental studies have been high in quantity and quality, and have covered all the key areas in the field of intensive care medicine: severe sepsis, multiple organ failure, transport and metabolism of oxygen, systemic and regional hemodynamic alterations (pulmonary, cerebral, hepato-splanchnic,...), metabolic disorders,... not forgetting issues associated with the complexities of medical ethics. The quality of care, clinical, scientific and personal, offered by the Department of Intensive Care at Erasme Hospital has earned it the high level reputation for which it is renowned in Belgium, and indeed worldwide. PMID- 12584938 TI - [The stomatology and maxillo-facial surgery department]. AB - Initially devoted to emergencies, the Department of Oral and Maxillofacial Surgery has grown considerably since 1979. The department now consists of a Maxillofacial Surgery Unit supplemented by several specialised Units in charge of oncology, orthodontics and orthognathic surgery, implantology and all dental specialities. The main themes of research developed in the department include the healing process of oral tissues, the teratogenic effects of various compounds on the cranio-facial region, the study of apoptosis, the role of heat shock proteins, and the management of craniofacial dysmorphosis. These studies have a direct impact on patient management. PMID- 12584939 TI - [The emergency department]. AB - After a brief history of Emergency Departments in Belgium, the evolution of the clinical activity and the main research topics developed in Erasme Hospital are developed, including the management of the department and of patients flows, and acute clinical toxicology. PMID- 12584940 TI - [The urology department]. AB - The Department of Urology of Erasme Hspital, University of Brussels, has participated to the development and implementation of new technologies that have significantly transformed the specialty in the last 25 years. The minimally invasive treatment of benign prostatic hypertrophia was developed by the use of Trans-Urethral Needle Ablation (TUNA) and new pharmacological treatments. Treatment of urinary stones by extracorporeal shockwaves lithotripsy and Endourology has allowed to avoid operating hundred of patients each year. In the field of prostate cancer, an original laparoscopic prostatectomy technique by the extraperitoneal approach was developed in our unit. The prevention of prostate cancer and the influence of various nutritional factors and its early diagnosis by the use of different markers have been the subject of numerous publications and have contributed to improve our knowledge in this field. Different prognostic factors of bladder cancer have been evaluated and in particular their place with the use of intravesical BCG. A better understanding of the different mechanisms involved in erectile dysfunction has been the subject of numerous studies during the last 20 years and the department is internationally recognised as a reference centre in this field. PMID- 12584941 TI - [The Erasme Foundation]. AB - Created in 1982 to encourage the scientific research in the Erasme Hospital, the Erasme Foundation has founded 174 research grants, has financially supported the building of the Cellular and Molecular Therapy Unit, and has organized more than 60 lectures in various biomedical fields. PMID- 12584942 TI - [Report on the union between the art of healing and modern science]. PMID- 12584943 TI - [Administrative role and position of the general physician in the follow-up of patients in home palliative care]. AB - The author names and discusses the different certificates the general practitioner can possibly deliver to a patient who needs palliative care at home in order to let him benefit of the care of a specialised team and of other social and financial advantages. He discusses afterwards the family practitioner's first rank position and his relations with the specialised palliative care teams (second line) and the specialised hospital services (third line). PMID- 12584944 TI - [Cochlear implantation, new prosthetic alternative in the management of the profoundly deaf child]. AB - Cochlear implants are new medical devices. In contrast to acoustic hearing aids which work by amplification, cochlear implants directly stimulate the hearing neurons via electrode implanted in the cochlear tympanic ramp. They are indicated in case of profound hearing deficiency or in cases of bilateral cophosis, congenital or acquired. Implanted early, they allow deaf children to improve their speech comprehension and their verbal expression considerably, thereby favouring social integration. A concurrent system of support including the use of visual-manual means of communication nevertheless remains vital for the first years. The frequency and duration of such support varies according to the individual. PMID- 12584945 TI - [NTBR (Not to Be Resuscitated) in 10 questions]. AB - The term "Not To Be Resuscitated" or "NTBR" is largely used in hospitals in the setting of therapeutic limitations. However, interpretations of this term are sometimes very different. The objective of this article is to define the exact meaning of "NTBR", namely, no resuscitation only in case of cardiac arrest. The rules for therapeutic limitation are also defined. PMID- 12584946 TI - [Drug treatment of epilepsy: update]. AB - Medical treatment of epilepsy has evolved during the last ten years. The "Epileptic Syndrome Classification" established in 1989 and new antiepileptic drugs account for these changes. This article recalls the principal rules of medical treatment of epilepsy and reviews new antiepileptic drugs available in Belgium. PMID- 12584947 TI - [Renaissance of thalidomide]. AB - Thalidomide comes back forty years after the discovery of its teratogenicity. Due to its antiangiogenic and immunomodulating properties, thalidomide is proposed in the treatment of multiple myeloma but also in the management of erythema nodosus leprosis, cutaneous lupus erythematosus and severe aphtosis. PMID- 12584949 TI - [Use of kohl and severe lead poisoning in Brussels]. AB - The authors report the observation of lead poisoning in a young Moroccan woman, which was caused by prolonged use of a khol made of lead sulphide. Khol is a black eye make-up used since Ancient Egypt. Clinically, this poisoning presented with abdominal crampoid pain, encephalopathy (manifested as anxiety and irritability), a Burtonian gingival border and microcytic sideropenic anaemia. Emergency chelate treatment permitted to improve clinical state and to decrease blood levels (initial blood concentration: 490 micrograms/dl; concentration six weeks after treatment: 49 micrograms/dl). The interest of such a case is recognizing, in French speaking regions, of a public health problem, largely ignored, although real, considering the high lead concentrations found in a majority of kohls available in Brussels and this very old cultural practice among Muslim populations. PMID- 12584948 TI - [Pleural effusion]. AB - A Spanish man is diagnosed with a non-small cell lung cancer with pleural extension. A chemotherapy combining cisplatin and gemcitabine allows obtaining an excellent partial remission. A contralateral pleural effusion is noted in a context of weight loss and fever. The differential diagnosis of pleural effusion is discussed. PMID- 12584950 TI - [Obstetrics and gynecology--retrospect of 125 years in Basel]. AB - The development of the discipline Obstetrics and Gynecology is reflected by the medical history at the University of Basel. At the very beginning of academic obstetrics, the cantonal government realised the need of a more practical teaching completing the tradition of theoretical lecturing (1865). Obstetrics was (and still is) a typical example for the turn from theoretical to hands-on teaching in medicine. At the time, when the independent chair of obstetrics at the University of Basel was founded (1867, Bischoff [1841-1892]), the fight against puerperal fever was the most urgent aim. The battle against puerperal fever was ultimately won around the turn of the 19th to the 20th century (von Herff [1856-1916]) but not much earlier. Step by step the gynecology grew out of the classical surgery where it has been located previously. Surgically skilled obstetricians like Fehling [1847-1925] and Bumm [1868-1925] have enhanced that development. Clinical research started with better information about the lying-in patients (Fehling, Bumm, Labhardt). Bacteriology in close connection with obstetrics was seen important (Bumm). Some decades later, clinical laboratories (clinical chemistry) were founded accompanying the diagnostic approach to various disorders of pregnancy (Labhardt [1874-1949]). Exfoliative cytology became mandatory since 1950 and this was the start to screening programs for early diagnosis of cervical cancer (Koller [1899-1985]). The family planning exploded in the years from 1965 on as the pill has been available, replacing sterilisation procedures (Labhardt). Endocrinology of pregnancy and cycle needed new biochemical methods. The laboratories were installed. Responding to the demands of a growing discipline, the building was enlarged and adapted in three major steps: 1896 new "Frauenspital", 1928 first enlargement, 1956-1968 constructing of two more storeys, of new surgical theatres and an entire tract for routine and scientific laboratories filling now the area between Klingelberg-, Schanzen- and Spitalstrasse. The hospital harboured in its peak time (1968-1970) more than 300 beds. In 1959 the hospital staff took care of 5000 deliveries (Koller). Fetal monitoring was introduced 1962, being the starting point for the development of "fetal medicine" in Basel. At that time, Otto Kaser [1899-1995] was chairman. He was appreciated as one of the most brilliant pelvic surgeons in Europe. Soon, the surgery for breast cancer was added to the classical gynecologic surgery. At the same time, ultrasound started to dramatically influence diagnostic procedures in pregnancy. In the eighties, in-vitro-fertilization and embryo-transfer improved the possibilities to overcome unwanted childlessness (Ludwig [*1929]) and supplemented endocrinology. The scientific spectre spread heavily to reproductive medicine since then, in particular to the molecular biology and research on stem cells, which was introduced by the present chairman (Holzgreve [*1955]). He has founded several research groups concentrating on the retrieval of floating fetal cells from cord and maternal blood, on other topics in fetal cell research, prenatal diagnosis and in ultrasound. PMID- 12584951 TI - [State of the art prenatal care from the Swiss viewpoint]. AB - Prenatal care has significantly reduced perinatal and maternal mortality. Screening for maternal disease allows us to reduce or to prevent an unfavourable fetal or obstetrical outcome. Prenatal care should start with a first preconceptional visit. Folic acid intake is recommended for all reproductive-age women who are capable of becoming pregnant. The fetal nuchal translucency measurement has revolutionized prenatal care as a non-invasive, effective screening for chromosomal abnormalities and other diseases of the fetus. Vertical transmission of infections has to be prevented if possible. As an example caesarean section in combination with antiretroviral therapy reduces the transmission of HIV significantly. Screening for sexually transmitted diseases (STD) remains important as at present the incidence of STD is increasing again. In this short review on prenatal care as it is done in Switzerland, we try to enlighten its most important aspects. For the patients and your own benefit as a physician it is important to follow guidelines, although of course each patient has to be treated individually. PMID- 12584952 TI - [Current aspects of labor induction]. AB - Induction of labor is one of the most important means for therapeutic intervention in modern obstetrics. The aim of labor induction is to achieve a better perinatal result for mother and baby as compared to expectative management. Different methods for induction include administration of oxytocin or prostaglandins, amniotomy, and mechanical means of cervical dilatation. The success of the labor induction depends primarily on the readiness of the uterus to go into labor, and the method used for induction. If the cervical ripeness is very advanced, induction with amniotomy and oxytocin seems beneficial. However if the cervix is not yet ready, intravaginal or intracervical prostaglandins are more promising. Until recently, prostaglandins E2 are used in the first line. Now, the prostaglandin E1-analogon misoprostol is also increasingly used. As a rule, induction of labor should be performed as an inpatient procedure in order to be able to provide the surveillance for maternal and fetal safety. PMID- 12584953 TI - [Ultrasound cervix measurement]. AB - The sonographic assessment of cervical length is a well established method for pregnant women with an increased risk of preterm delivery. Even in asymptomatic patients, but with specific risks for preterm delivery, a selected screening has become successful. Up to now a screening in the general population is not recommended, as the sensitivity and specificity in this group are quite low. Therapeutic interventions or preventions are more efficient and under objective criteria with the use of cervical length measurement. The further development will be a combination of sonographic, biophysical and biochemical markers. This includes the multiple causes for preterm delivery and pronounces the central role of sonographic assessment of the zervix. For induction of labour at term the results of cervical length assessment are promising. The interval between priming and delivery and the success rate for a vaginal delivery within 24 hours can be predicted more precisely. It optimises the organisation of a busy delivery unit and motivates the pregnant woman. PMID- 12584954 TI - [Elective cesarean section--delivery mode of the future or social misconception?]. AB - Over the past 20 years, the rate of Cesarean section has undergone a marked increase on a global level and it appears unlikely that this trend will be reversed in the near future. This fact has also raised a number of questions regarding the growing preference for elective Cesarean section following a complication-free pregnancy and in the absence of medical indications. Both, the mortality rate of carefully prepared procedures of this kind, as well as the attending morbidity rate, have been successfully reduced in recent years owing to improvements in section technique, causing certain of the maternal and fetal risks accompanying vaginal delivery to come under closer scrutiny than before. The potential damage to the perineum during birth--and the impairment of the parturient's sexuality this may cause--in conjunction with the fact that vaginal delivery continues to expose the infant to certain risks which cannot be ruled out entirely, has made the choice between vaginal delivery and Cesarean section increasingly difficult. The argument most frequently cited in favour of vaginal delivery concerns the unimpaired birth experience it offers. Many women, however, do not consider this a high priority. In keeping with the greater importance currently being assigned to patients' preferences, the authors support the view that the parturient herself should be enabled to decide what level of risk is acceptable to her, including stating her preference for a delivery by Cesarean section. It cannot be stressed enough that, in this context, comprehensive information and rigorous documentation are indispensable requirements. PMID- 12584955 TI - [Anesthesia and analgesia in obstetrics]. AB - Anesthesia in obstetrics includes the medical attendance of women in the delivery room as well as giving anesthesia for cesarean sections in the operating room. Over the last years the treatment of labor pain with epidural anesthesia has been modified. Whereas a couple of years ago local anesthetics were used almost exclusively, the recent trend goes toward a combination of local anesthetics with opioids. Using this combination the total amount of local anesthetic can be greatly reduced, whilst maintaining most of the motor function during labor. There is evidence the combination of local anesthetics with opioids can reduce the number of operative vaginal deliveries such as vacuum extraction or forceps. The systemic application of opioids remains unaffected by the local application and its significance is unaltered, even though the effectiveness compared to epidural application in managing labor pain is far inferior. Opioids applied systemically often have an unwanted sedative effect in the mother and have the potential for respiratory depression in the newborn. The question, whether epidural anesthesia increases the frequency of cesarean sections remains to be answered. Studies so far present discrepant results and do not show a causal relationship between the use of epidural anesthesia and increase in cesarean section rate. Regarding cesarean sections, there has been a trend in the operative field over the last years towards the use of regional anesthesia. This is the consequence of the fact that anesthesia related mortality during cesarean sections is still mainly due to hypoxia and aspiration during induction of anesthesia. The advantage of spinal anesthesia over epidural anesthesia is faster onset, more reliable sensitive block and a lower failure rate. Downside of the use of spinal anesthesia is a higher incidence of hypotension in the mother, which, however, is not a serious complication if treated adequately. Regarding the continuous application of local anesthetics via a spinal catheter, no definite statements towards benefits compared to other regional techniques can be made due to the lack of adequate amount of patient studies. Theoretically this method seems advantageous as it allows to adjust the administration of local anesthetics and opioids to the individual needs in a very refined way. Emergency situations, such as emergency cesarean sections, life threatening hemorrhage, eclampsia, and HELLP syndrome, are the main risks of the anesthetic practice in obstetrics. Their beneficial outcome is highly dependent on the coordination of logistic problems, good communication and coordination within an interdisciplinary team of obstetricians and anesthesiologists. PMID- 12584956 TI - [Breech presentation in obstetrics in Germany--an assessment of current status]. AB - Nowadays, the CS seems to be the most favourite way of delivery in case of breech presentation. Our opinion is that the vaginal delivery has no disadvantage for the physical and mental development of the newborns [11, 12]. The outcome depends on the set-up and organisation of each department. The most important condition to reach a good outcome by breech presentation is a well-established practical training of the whole team, a careful risk selection and a well structured perinatal centre. In our opinion the CS is the second choice to deliver a pregnant woman with a breech presentation at term. High CS-rates as well as a increased perinatal morbidity at vaginal breech delivery are a sign for insufficient training of the staff and insufficient organisation in small departments not being perinatal centres. Not the breech presentation is the real risk factor, but the unqualified Obstetrician and the insufficient set-up in the most of OB/Gyn-departments. We have a very high ethic responsibility against young women resultating from our profession. It's our duty to decide and recommend the best way to save the wellbeing of the mother and the unborn child. It's also our duty to avoid surgical manoeuvers resulting from economic reasons. Therefore a complete, sufficient practical training concerning breech delivery technics is absolutely needed. In future the answer of the question: "Delivery of breech presentation--vaginally or through CS?" will depend on, if we will be able to change this politic and professional undesirable trend. PMID- 12584957 TI - [Multiple pregnancy]. AB - The incidence of multiple pregnancies has increased considerably over the past 20 years in the western world. The number of twin pregnancies has increased by approximately 25% and the rate of triplet and higher order multiple pregnancies has more than doubled. This is due to the use of ovarian hyperstimulation and assisted reproduction techniques, and to the increasing maternal age. Multiple pregnancy can be considered as the most important adverse outcome of infertility treatment. The main determinant for outcome and management of multiple pregnancies is the chorionicity. This can be determined by ultrasound until 14 weeks of gestation. Twenty percent of all twin pregnancies are monochorionic, and 15% of these develop severe twin to twin transfusion syndrome, which is best treated by LASER separation of the communicating blood vessels. Obstetric complications associated with multiple gestation also occur more frequently in monochorionic twins. They include increased incidence of preterm labor, intrauterine growth restriction and assisted or surgical delivery. Neonatal problems include low birthweight and increased prevalence of congenital malformations. Overall, there has been a decrease in neonatal mortality of twins and triplets over the past 10 years, which is mainly due to the enormous advances in neonatal intensive care. PMID- 12584958 TI - [Alternative obstetrics: bed, chair or tub? Have alternative birthing methods become established?]. AB - As a reaction to the technology craze that occurred in obstetrics, the pendulum swung toward alternative obstetrics with alternative birthing methods. Instead of the classical bed birth on a narrow bed, birth on a wide bed and births using the Maya birthing stool and waterbirths became established as alternative birthing methods. Using the example of the Women's Clinic of the Cantonal Hospital of Frauenfeld, this study shows how alternative obstetrical medicine was introduced and could be integrated into traditional obstetrics. The goal of the Frauenfeld obstetrics is to combine the parturient's personal wishes and desires with classical obstetrics. Using a quality analysis and prospective observational study that has been on-going since 1991, it could be shown that alternative birthing methods are just as safe for the mother and child as is the classical bed birth on the narrow bed. In addition to ensuring the safety for the mother and child, alternative obstetrics is responsible for the attitude that the birth be considered an unforgettable and beautiful experience for the parturient, which is a definite advantage for women. Alternative birthing methods have become established because they have been able, using a high technical standard, to combine safety for the mother and the child with a humane side of obstetrics that evokes a nurturing and warm atmosphere. PMID- 12584959 TI - [Decreasing the rate of premature delivery with phytotherapy--results from general practice]. AB - The paper discusses analysis of data obtained from a 23 years obstetric practice period with special reference to tocolysis and premature delivery. 253 patients were subjected to tocolysis out of the 1622 deliveries. There were 29 premature deliveries which gives a 1.8% rate. Major breakthrough in the success of tocolytic treatment could be attributed to the introduction and use of the phytopharmacon Bryophyllum. In the period between 1977 to 1983--the Partusisten tocolytic treatment period--the frequency has been reduced to 6.2%. Following the introduction of Bryophyllum this frequency has been reduced to 1.07%. Majority of the cases responded well to the outpatient oral Bryophyllum tocolysis combined with a part-time bedrest. The above described preventive measure in turn would result in a significant decrease of the expenses. PMID- 12584960 TI - Is there a "representative" sample? PMID- 12584961 TI - Supporting older adults living with multiple chronic conditions. AB - This qualitative study was conducted using focus groups to explore the strategies commonly employed by older adults (N = 37) to manage multiple chronic conditions. Key strategies identified were relating with health care providers, medicating, exercising, changing dietary patterns, seeking information, relying on spirituality and/or religion, and engaging in life. Although social support was not mentioned as a discrete strategy, the participants' social networks were embedded in all of the categories. This study supports building a partnership of care in which nurses and other health care professionals function in supportive and educative roles to enhance the older person's lifelong self-care management and ability to stay in control of multiple chronic health conditions. PMID- 12584962 TI - Parents' reactions to conflict with health care providers. AB - The purpose of this phenomenological study was to examine parents' reactions to conflict with health care providers and the outcomes of the conflicts. This study builds on previous research that confirmed the occurrence and sources of conflict among these informants. Seventeen parents whose children had a diagnosis of cancer were interviewed in this study. A priori coding was used to identify the specific strategies for dealing with conflict proposed by Rubin and associates. Ginsburg's approach to narrative analysis was used to examine parent's stories. Findings were that parents used the strategies of problem solving, yielding, withdrawing, inaction, and contending identified by Rubin and associates in reaction to conflict with health care providers. Following episodes of contending, many parents in the study reframed the conflict and experienced transformations or turning points, resolving to become more effective advocates for their children. A few of these advocates proceeded to become community activist. PMID- 12584963 TI - The experience of being an older staff nurse. AB - The nursing workforce is aging at an unprecedented rate, yet we know very little about the experiences of older staff nurses. A qualitative design with purposeful sampling was used to describe the experience of being an older staff nurse. Data were obtained through in-depth interviews with 11 staff nurses over the age of 55 who were employed at least part-time in six hospitals located in the southeastern United States. Data analysis was guided by a feminist perspective utilizing the voice-centered relational method developed by Brown and Gilligan. Study findings demonstrate that older nurses are working because they continue to care, despite the stressors of intergenerational conflict with younger nurses, less respect from patients and families, and inequity in pay. Older nurses are confident in their abilities and are capable of meeting the demands of hospital nursing. PMID- 12584964 TI - Lack of awareness and nonadherence in schizophrenia. AB - This cross-sectional study examines whether lack of awareness of symptoms is a predictor for nonadherence. The study consecutively recruited 134 participants with Diagnostic and Statistical Manual of Mental Disorders, 4th edition, diagnoses of schizophrenia spectrum disorders at acute psychiatric units. Outcome was measured by a combination of self-reports, utilizing the Rating on Medication Influences and medical records. The univariate analysis showed that homelessness was a predictor for nonadherence (p = .03). The subsequent logistic regression showed that the multidimension symptom lack of awareness score was significantly associated with nonadherence to psychotropic medications (odds ratio = 0.59, 95% Confidence Interval = 0.35, 0.96). Other clinical and demographic variables as well as subjective responses were not significant predictors of nonadherence. PMID- 12584965 TI - Resourcefulness and self-care in pregnant women with HIV. AB - The human immunodeficiency virus (HIV) has caused suffering in the lives of humans worldwide. Pregnant Thai women now constitute the fastest-growing segment of individuals diagnosed with HIV/AIDS. Few studies have looked at self-care or resourcefulness among pregnant women with HIV. Using Rosenbaum's theory of learned resourcefulness, this study examined the direct effects of depression and resourcefulness on prenatal self-care as well as the mediating effects of resourcefulness on depression and self-care. A model testing study with 153 pregnant Thai women compared the effects of depression and resourcefulness on prenatal self-care in HIV-positive and HIV-negative groups. Regression analyses indicated direct effects of depression and resourcefulness on prenatal self-care. The effect of depression on prenatal self-care was mediated by resourcefulness. HIV status did not predict prenatal self-care. The findings on the relationships of depression, resourcefulness, and prenatal self-care can help nurses provide effective services to pregnant Thai women, including counseling on self-care. PMID- 12584966 TI - Spirituality and health in older Thai persons in the United States. AB - Health and spiritual phenomena are viewed differently depending on the cultural perspective. This study describes spirituality as perceived and experienced by older Thai persons. Specific aims were to describe how spirituality helped older Thai persons maintain their health and to describe what they valued most as they aged. A qualitative, descriptive study informed by ethnographic methods was utilized. The sample included 9 older Thai persons from an urban U.S. community. Data were generated using ethnographic interviews and participant observation. Data analysis incorporated coding, categorizing, and theme development. Rigor was guided by Lincoln and Guba. Five major themes emerged from the data: Connecting with spiritual resources provided comfort and peace, finding harmony through a healthy mind and body, living a valuable life, valuing tranquil relationships with family and friends, and experiencing meaning and confidence in death. For these Thai participants, health and spirituality coexisted and were linked to all of life. PMID- 12584967 TI - Genetics, genomics and molecular medicine. AB - Genomics and the Human Genome Project are creating important changes in medical care and radiologic practice. Readers of this article will: Recall that deoxyribonucleic acid (DNA) is the information-containing molecule. Understand the relationship between DNA and protein synthesis. Learn that changes in DNA encoded proteins produce pathogenesis. Distinguish between genomics and proteomics. Describe the Human Genome Project. Define and give examples of molecular medicine and its imaging applications. Discuss how genomics, genetic engineering and the Human Genome Project may influence radiologic practice. PMID- 12584968 TI - MQSA update. AB - Since the MQSA regulations took effect in 1999, several amendments have been enacted, most notably new stringent equipment guidelines that became effective on October 28, 2002. After reading this review of the amended final regulations, readers will: Be familiar with the amended final equipment and quality assurance regulations. Understand policy guidance help regarding the new regulations. Understand new policy guidance for radiologic technologists. Know the basics of MQSA's impact on providers, patients and mammography quality and reasons for facility citations since release of the final regulations. Know where to go for MQSA information and guidance resources. PMID- 12584969 TI - Saying goodbye to yesterday. PMID- 12584970 TI - To scan or not to scan. PMID- 12584971 TI - The importance of the laboratory class. PMID- 12584972 TI - Patient page. Skull radiography. PMID- 12584973 TI - Median and ulnar nerve compression at the wrist caused by anomalous muscles. AB - Compression of the median and ulnar nerves at the wrist is frequently encountered. Carpal tunnel syndrome usually occurs without any obvious extrinsic cause; several cases have however been reported caused by anomalous or hypertrophic muscles. A survey of the literature shows that compression neuropathy of the median nerve has been reported in relation with anomalies affecting three muscles: the first (or second) lumbrical, the palmaris longus and its anatomic variants and the superficial flexor of the index finger. In the ulnar tunnel the situation is thoroughly different: so-called idiopathic ulnar tunnel syndrome is rare and an extrinsic compressing structure can usually be disclosed. Anomalous muscles belong to the palmaris longus/abductor digiti minimi group; the flexor carpi ulnaris is sometimes involved. One can suspect the presence of such an anomalous muscle when the compression syndrome concerns a patient who is not within the "usual" age group with symptoms initiated or aggravated by physical exercise. PMID- 12584974 TI - [Orthopedic immobilization]. AB - The classical plaster bandage was devised in the mid 19th century. Until recently, osteoarticular trauma has been treated mostly by plaster cast immobilisation using plaster of Paris. Synthetic materials have been introduced on the market place in the seventies, but they have not superseded the traditional plaster of Paris. The more recent thermoplastic materials are used to make splints and orthoses, particularly at the wrist and hand. The present review of the literature confirms that synthetic materials present better physical and mechanical properties than the traditional plaster of Paris. In addition, they are lighter, they are more resistant to humidity, they are more radiotransparent and they generate less dust when removed. However, they are less malleable and cause higher pressure in case of limb edema. Plaster of Paris therefore remains indicated in the acute posttraumatic or postoperative period. This material is also cheaper, but the pecuniary benefit is limited for several reasons, particularly because plaster of Paris is associated with a higher rate of cast replacement. PMID- 12584975 TI - Operative treatment of humeral shaft fractures. The Leuven experience. AB - For many years, plating has proved to be a reliable method for the fixation of fractures of the humeral shaft. In the early nineties however, intramedullary devices became increasingly popular for fractures of the humeral shaft. This was based on a global tendency towards minimally invasive surgery, and the attractiveness of the relatively simple procedure and potentially lower complication rate of intramedullary nailing, which had proved to be successful in osteosynthesis of the lower limb, However, until now there is no consensus in the literature as to which device is preferable for different indications. We reviewed 161 patients, operatively treated for a fracture of the humeral shaft in our department between 1986 and 1999. Our experience shows in most indications a higher union rate, better functional results and a lower reoperation rate after plate and screw fixation. In addition, even though plating requires a more technically demanding procedure, in experienced hands, it gave rise to fewer iatrogenic fractures, and fewer persisting pain problems. We recommend the use of plate and screws as primary treatment in all operative indications, except for pathological fractures, very obese patients, and open fractures. PMID- 12584976 TI - Treatment of humeral diaphyseal nonunions by interlocked nailing and autologous bone grafting. AB - Twenty seven patients with humeral diaphyseal nonunion treated with interlocked nailing and autogenous bone grafting were retrospectively assessed. The mean age was 37 years (28 to 59 years). There were 16 women and 11 men. An average of 11 months (8 to 36 months) had elapsed between the initial trauma or treatment and presentation in our clinic. The nonunion site was in the distal third of the humerus in 18 patients (66%) and in the middle third in 9 (34%). Seventeen (73%) had hypertrophic and 10 (37%) atrophic nonunion. Patients were treated with locked intramedullary nailing and autogenous cancellous bone grafting. The mean follow-up was 42 months (28 to 62 months), and the mean time to union was 4.8 months (2.5 to 11 months). Three patients developed superficial infection. Union was achieved in all cases but one, in which there was multiple nerve injury. According to Steward and Hundley's scoring, results were good in 24 patients, fair in 2, and poor in one. Interlocked nailing and autologous bone grafting has a good rate of union provided nails of appropriate diameter are used and distal and proximal locking is performed correctly. We think that its low rate of infection, low risk of injury to the radial nerve, and low requirement for soft tissue dissection make it a suitable choice in the treatment of nonunions of the humeral diaphysis. PMID- 12584977 TI - [Lesions of the distal radio-ulnar joint associated with isolated fractures of the radial shaft]. AB - The authors have performed a retrospective study of 49 Galeazzi fractures treated between 1990 and 1998. This lesion is considered rare because it is often misdiagnosed as an isolated fracture of the radius. The mean age of the patients was 31 years. There was a male predominance with a sex ratio of 4/1. Road traffic accidents were the main etiology (45%). Galeazzi fracture type III in Mansat's classification represented 53%, followed by type II (33%), type I (8%) and equivalents of Galeazzi fracture (6%). The treatment was surgical in all cases. The radial fracture was internally fixed with a plate. Reduction of the distal radio-ulnar instability, achieved by manipulation, was maintained with radio ulnar pin fixation in 53% and with plaster cast immobilization 45%. The results, evaluated according to Mikic's criteria were excellent in 87%. The prognosis of Galeazzi's fracture depends mainly on the initial treatment of the lesions of the distal radio-ulnar joint, which require for their diagnosis a meticulous clinical evaluation and a good radiological analysis. PMID- 12584978 TI - [Vitamin C and prevention of reflex sympathetic dystrophy following surgical management of distal radius fractures]. AB - Reflex sympathetic dystrophy is a major complication following surgical treatment of fractures of the distal radius. Its pathogenesis is related to lipid peroxidation which damages vascular endothelial cells, increasing capillary permeability. Vitamin C is a natural antioxidant. The authors have made a comparative study of two groups of patients with isolated closed displaced fractures of the distal radius, which were reduced and stabilized by intrafocal pinning. Group 1 included 100 patients who were treated from 1995 until 1998 and who did not receive any vitamin C supplementation; group 2 included 95 patients who were treated from 1999 to 2002 and who received daily administration of one gram vitamin C orally during 45 days, starting on the day of fracture. The incidence of reflex sympathetic dystrophy was five time times lower in group 2 (2.1% versus 10%). This is in line with previous observations and lends credit to the value of vitamin C administration as a prophylactic measure to prevent the occurrence of reflex sympathetic dystrophy in patients who undergo surgical treatment of a displaced fracture of the distal radius. PMID- 12584979 TI - Osteochondritis dissecans of the femoral head in Perthes disease: a cause for concern? AB - Osteochondritis dissecans (OCD) and loose body formation are rare following Perthes' disease. We have reviewed the literature about clinical presentation, treatment and outcome and added a further three cases of the condition. Cases mentioned in the literature were poorly documented. We feel that a thorough documentation should be carried out as soon as the diagnosis is made. Conservative treatment should be given when the disability is moderate. The loose body should only be removed surgically when it is mobile, when it bulges into the joint space or when there are signs of early arthritis. PMID- 12584980 TI - The effectiveness of dual offset stems in restoring offset during total hip replacement. AB - Commonly, total hip prostheses have had a higher neck-shaft angle than the host bone and thus a tendency to reduce the femoral offset. Restoration of the femoral offset may be important as it has been shown to enhance hip stability and to improve the range and strength of abduction. The purpose of this study was to determine which of two designs was best able to restore femoral offset in comparison to the contralateral normal hip. Two hundred and two primary total hip patients were included in a radiographic study. Measurements were taken from a postoperative anteroposterior radiograph of the pelvis. The Synergy femoral component with a more varus neck-shaft angle of 131 degrees and a standard or high offset option tended to restore the femoral offset more reliably than did the Mallory-Head femoral component with a neck shaft angle of 135 degrees. PMID- 12584981 TI - Clinical outcome after treatment of infected primary total knee arthroplasty. AB - Twenty-six consecutive cases of infected primary total knee arthroplasties were treated at our institution from 1989 through 2000. Eleven patients had debridement and irrigation performed within 2 months of index arthroplasty or hematogenous spread; only one infection was eradicated. Twenty-five patients had their prostheses removed; 17 had two-stage revision arthroplasty, following which infection was eradicated in 15; one had a permanent spacer, 7 had arthrodesis (following failed revision arthroplasty in one) and 2 had a femur amputation (following failed revision arthroplasty in one) at follow-up of mean 24 months. Infections were cured equally well with revision arthroplasty and arthrodesis. Among the 15 patients who ended up with revision arthroplasty, 11 had a better range of motion compared to the index arthroplasty, but 8 had daily pain. We present our treatment protocol, which eradicated 15/17 (88%) infections in patients treated with two-stage revision arthroplasty. PMID- 12584982 TI - Dynamic external fixation of comminuted intra-articular fractures of the distal tibia (type C pilon fractures). AB - We report 26 patients with 28 type C3, distal intraarticular tibial (pilon) fractures treated by dynamic external fixation. Follow-up was at least two years, and the results (subjective and objective) were classified according to the Ovadia system. The mean to fracture union was 14 weeks (range: 12 to 20 weeks). There were three cases with angulation deformity (from 7 degrees to 20 degrees). There were no cases with nonunion or deep infection. Based on these results, this treatment with closed reduction and dynamic external fixation allowing early motion appears as a suitable method for treatment of comminuted intraarticular tibial pilon fractures. PMID- 12584983 TI - [Ankle instability with involvement of the subtalar joint demonstrated by MRI. Results with the Castaing procedure in 45 cases]. AB - The authors report 46 cases of instability of the hindfoot involving the subtalar joint, with or without combined involvement of the tibiotalar ligaments, which were treated using Castaing's technique of ligament plasty between 1988 and 1999. Preoperatively, the patients complained of instability, recurrent sprains and pain. A tarsal sinus syndrome was present in 39%. Forty six patients underwent NMR examination which demonstrated capsuloligamentous lesions in every case. The results were evaluated using Kitaoka's score. With an average follow-up of 5.7 years after operation, instability was no more present in 80% and 63% were painfree. Clinical examination showed a reduction in the range of inversion of the hindfoot in 43%, with values between 50 and 70% of the contralateral foot, but without any significant clinical repercussion. Incipient osteoarthritis was noted on xrays in three patients. The overall results were very good in 82%, fair in 11% and poor in 7%. Eighty seven percent of the patients were satisfied with the result of the operation. The authors found a significant correlation between fair or poor results and a body mass index above 26 kg/m2 or constitutional laxity. Besides, results got worse as the time interval between the first sprain and the operative treatment increased. Comparison of these results with those of other techniques shows that they are essentially similar. Ligament plasty using Castaing's technique is a reliable operation but direct repair of ruptured ligaments at the subtalar joint should be preferred for primary treatment, leaving the possibility for secondary ligament plasty if necessary. PMID- 12584984 TI - [Traumatic dislocation of the radial head associated with ipsilateral fracture of the radial shaft: a case report]. AB - Ipsilateral radial head dislocation with radial shaft fracture is a rare injury. A few cases have been reported in the literature. We report the case of a 39-year old man who presented with a radial shaft fracture and ipsilateral dislocation of the radial head caused by hyperpronation trauma of the forearm with hyperflexion of the elbow. The dislocation was reduced by manipulation after open reduction of the radial shaft fracture, which was then fixed with a plate. The clinical and radiological outcome was good with 2 years follow-up. PMID- 12584985 TI - Osteoblastoma of the carpal scaphoid frequency and treatment. AB - A rare case of osteoblastoma of the carpal scaphoid is presented. Review of the literature revealed two more cases that have been previously reported. The authors present the diagnostic difficulties and the treatment which was applied: proximal row carpectomy and tendon transfers for wrist stabilization. Treatment options, according to the literature, also include scaphoidectomy and tumor curettage with bone grafting. PMID- 12584986 TI - [Fatigue fracture of the femoral neck in an HIV-positive female patient on antiretroviral therapy]. AB - The authors report the case of an HIV-infected patient on highly active antiretroviral therapy (HAART) who presented with spontaneous fracture of the right femoral neck with avascular necrosis, probably related with her HIV status and HAART, and who was treated by non-cemented arthroplasty. PMID- 12584987 TI - Traumatic hernia of the abdominal wall after pelvic and acetabular fracture: a case report. AB - Traumatic hernia of the abdominal wall is a rare and easily missed injury in the presence of major pelvic and abdominal lesions. We present a radiographically documented case of combined pelvic and acetabular fracture with a major contralateral traumatic hernia with avulsion of the internal oblique, the external oblique and the transverse abdominal muscles diagnosed four months after the initial trauma. To our knowledge no similar case has been described in the current literature. PMID- 12584988 TI - Intrapelvic mass in a young woman with an uncemented hip arthroplasty. AB - Based on a case report in which wear of the polyethylene liner in an uncemented hip prosthesis led to the development of an intrapelvic mass, the pathogenesis, diagnosis and treatment are described. Characteristic osseous and soft tissue reactions caused by release of polyethylene particles to periarticular tissue are discussed as well as the differential diagnosis with infectious and malignant conditions. In the case treated, the differential diagnosis was also made with respect to gynecological pathology. PMID- 12584989 TI - [An unusual localization of solitary bone cyst in a child: the lumbar spine]. AB - Solitary bone cysts involving the spine in children are extremely rare. We report such a lesion in a 7-year-old girl who presented with an expansible, bubbly cystic lesion of the third lumbar vertebra. The intraoperative findings and histologic examination indicated the diagnosis of a solitary bone cyst. She was treated successfully with curettage and bone grafting. PMID- 12584990 TI - [Endocrine aspects of diagnosis and therapy of pheochromocytoma]. PMID- 12584992 TI - [Personal experience in diagnosis and localization of pheochromocytoma]. AB - INTRODUCTION: Pheochromocytomas are most commonly tumours of adrenal medullary origin. Pheochromocytoma by definition produces and secretes catecholamines. Similar tumours that do not secrete active substances of any kind are called non functioning paragangliomas. The hallmark clinical manifestation of pheochromocytoma is hypertension accompanied with various signs and symptoms in excess of catecholamines or other bioactive substances. The early diagnosis of pheochromocytoma is important not only because it offers the possibility of curing hypertension but also because unrecognised pheochromocytoma is a potentially lethal condition. The aim of this article is to stress the specify of the clinical finding, diagnostical values of the laboratory tests and possibilities of morphological localizing techniques in a series of 98 patients with surgically proven pheochromocytoma. RESULTS: Over the period from 1954 to 2002 pheochromocytoma was diagnosed and surgically treated in 98 patients. The diagnosis was confirmed at operation except in patients who refused operation or continued the examination in other Clinical wards. There were 59 females and 48 males (F:M = 1.23:1), the age ranged from 7 to 64 years with the pick incidence in the second and third decades of life in males and the third and fourth decades of life in females. The basic clinical characteristic was hypertension which was found in 94% of patients with an approximately equal frequency of fixed and paroxysmal hypertension cases. The most often accompanning manifestations were headache (62%), perspiration (61%) and palpitations (65%). A high level of vanyl mandelic acid (VMA) and free catecholamines in 24-hour urine collection confirmed the diagnosis in 94% of cases. In boderline cases we performed dynamic tests, the most relevant among them being the test with phentolamin. It was positive in 95% of patients. Retropneumothomography contributed to a successful localisation of tumour in 83% of cases. Computed tomography (CT) was performed in 69 patients and was positive in 97% of them. Magnetic resonance imaging (MRI) localized the tumour in all 16 patient in whom it was performed. The whole body MIBG-J-131 (metaiodobenzylguanidine) scanes were positive in 92% (45/49) and false negative in the remainder of 8% (4/49) of cases. Selective angiography was performed in 40 patients and in all it was positive. DISCUSSION: Although pheochromocytomas were among the first recognized adrenal tumours, the prompt and safe diagnosis is mandatory up to date. The average annual incidence has been estimated by several epidemiologic studies to range from 0.8 to between 1.55 and 2.1 million persons per year. It is reported that it is curable cause of hypertension in 0.1% to 1% of cases. Pheochromocytoma has been classified as a "10% tumour" because various studies have shown that each of the characteristics mentioned bellow occurs with a frequency of approximately 10%: bilateral, extra-adrenal, multiple, malignant, familial and occurring in children. Our series of patients has a similar distribution: pheochromocytoma was in 9.2% of patients extra-adrenal, in 7.1% bilateral, in 9.2% multiple and in 4.08% malignant. Hypertension was the constant finding in 94% of our patients. Three clinical patterns of hypertenson have been observed. The first is paroxysmal hypertension, and the others are fixed or combinations of fixed and paroxysmal hypertension. According to our experience there were the equal incidence of all forms of hypertension. We noticed, like others, when the triad of headache, sweating and palpitations is accompanied by hypertension, the diagnosis of pheochromocytoma can be made with specify and sensitivity over 93%. In absence of this finding the diagnosis can be excluded with certainty of 99%. As a specify of the clinical finding, we mention two patients with manifestations of hypercorticism, two patients with pheochromocytoma of the urinary bladder, and four with MEN syndrome (one with MEN 2A and three with MEN 2B). For confirming the diagnosis the most relevant laboratory test was the higher level of VMA and free catecholamines in 24-hour urine collections. Once pheochromocytoma has been diagnosed by biochemical analyses, the anatomic location of the tumour or tumors must be determined. Currently, the best approach is to obtain MIBG-J-131 scan and then to perform MRI or CT of the abdomen and other areas identified on MIBG scan in order to provide more accurate spatial information. With this approach the great majority of pheochromocytomas can be localized. CONCLUSION: There is no classic picture, no stereotype for pheochromocytoma, although the history and physical finding are helpful. Patients come to the clinician in a variety of ways and settings. They may have classic attacks of hypertension accompanied with headache, perspiration and paplpitations or they may have identical symptoms and physical findings as the patients with primary hypertension. On the other hand, they may have signs and symptoms of diabetes mellitus, hyperthyroidism, hypercalcaemia, congestive heart failure, myocardial infarction, malignant hypertension or a variety of other conditions. Rarely, they have no complaints at all. Once the diagnosis was made, spatial localizing of the tumour or tumours, and surgical treatment are necessary. Unrecognized disease may be fatal. PMID- 12584991 TI - [Pheochromocytoma--pathohistologic and immunohistochemical aspects]. AB - INTRODUCTION: Pheochromocytoma originates in chromaffin cells of the adrenal medulla. Its incidence is similar in both sexes and most frequent between the ages of thirty and fifty. Multiple and bilateral pheochromocytomas constitute 5 to 10 percent of all cases. Pheochromocytoma occurs sporadically or is related to family syndromes such as: syndrome of multiple endocrine neoplasia--MEN IIA and IIB, neurofibromatosis (von Recklinghausen's disease), von Hippel-Lindau's disease, Sturge-Weber's syndrome, and tuberous sclerosis. Cases in a family usually occur at a younger age and are mostly bilateral and with more aggressive biological behaviour. MATERIAL AND METHODS: The aim of the study was to make histomorphological and immunohistochemical analyses of 52 pheochromocytomas. These cases are the surgical material from the Centre of Endocrine Surgery, Institute of Endocrinology, Diabetes, and Metabolic Disorders, Clinical Centre of Serbia, Belgrade, over the period from 1974 to 1997. Frozen and fixed sections, which were cut from paraffinembedded material and stained by both hematoxylin eosin and PAS, were used in order to make pathohistological diagnoses. The expression of chromogranin A, S-100 protein and ACTH was examined using the PAP method, while neuronspecific enolase (NSE), synaptophysin and neurofilament were examined by the APAAP method with appropriate antibodies (DAKO). RESULTS: The patients were between 4 and 65 years of age (average age 38.5) and there were 28 females (63.64%) and 16 males (36.36%). The largest pheochromocytoma had the diameter of 12 cm, and weight of pheochromocytomas in question was from 13.5 to 370 grams, the average weight being 83.4 grams. On gross examination, the tumours proved to be well-defined, either by fibrous capsule, or by adrenocortical tissue. The cross-sections of tumours were mainly of pale red-grayish colour, and showed numerous foci of necrosis, haemorrhage and cystic softening. Histological appearance of pheochromocytomas was with significant irregularities in shapes and dimensions of the cells and their patterns. Pheochromocytes were mostly of polygonal shape (45 cases, 86.54%), whereas in 7 cases (13.46%) fusiform cells were evident. Cells were arranged, either in trabeculae intermingled with thin walled sinusoids, or in small alveolae circumferenced by fibrovascular stroma. PAS positive hyaline globules were often present in the cell cytoplasm and also extracellularly. Cellular and nuclear pleomorphism, binuclear and multinuclear cells, as well as giant cells were evident in 35 (67.31%) pheochromocytomas. Mytotic figures were infrequent in 50 tumours (one to two on ten microscopic high power fields). Malignant pheochromocytomas (2 cases) metastasized to regional lymph nodes and liver, and lungs and bones, respectively. Pheochromocytomas were associated with MEN IIA syndrome in 6 patients (13.63%), all of whom had bilateral adrenalectomy, and in the period from two months to one year later underwent total thyroidectomy due to medullar thyroid carcinoma. Simultaneous occurrence of adrenal medullar hyperplasia and pheochromocytoma, i.e. the sequence: diffuse medullar hyperplasia--nodular medullar hyperplasia pheochromocytoma, was found in two patients with MEN IIA syndrome (33.33%). Hyperplastic nodule (1 cm in diameter) with discrete capsule and compression of surrounding adrenal tissue was considered to be a small pheochromocytoma. All the studied pheochromocytomas showed chromogranin A, NSE, and synaptophysin immunopositivity in nearly all tumour cells. Neurofilament positivity was focal and less intense. S-100 protein positive sustentacular cells were found in 29 of 32 pheochromocytomas included in immunohistochemical examinations. Incidence frequency of S-100 protein positive sustentacular cells was high in pheochromocytomas related to family syndromes and low in malignant pheochromocytomas. DISCUSSION: Numerous features of the described pheochromocytomas appear in the already published data, namely: age of occurrence, tumour weight, histological and immunohistochemical characteristics, frequent association of bilateral pheochromocytoma and MEN IIA, as well as the occurrence of adrenal medullar hyperplasia as precursor of pheochromocytoma. Immunohistochemical analysis has confirmed the importance of pan-neuroendocrine markers (chromogranin A, NSE, and synaptophysin) in pheochromocytoma diagnosing, whereas it has been that neurofilament was of less importance. High incidence frequency of S-100 protein positive sustentacular cells in pheochromocytomas related to family syndromes was also noted by other authors. CONCLUSION: Histomorphological features of benign and malignant pheochromocytomas may be similar. Histologic criteria for aggressive biological behaviour of pheochromocytoma include: insular pattern of growth, 3-5 mytotic figures on 10 microscopic high power fields and invasion of capsular lymphatics and blood vessels. Pan-neuroendocrine markers (chromogranin A, NSE, synaptophysin) may be useful in diagnosis of pheochromocytoma. Incidence frequency of S-100 protein positive sustentacular cells is high in pheochromocytomas related to family syndromes and low in malignant sporadic pheochromocytoma. PMID- 12584993 TI - [Nuclear medicine diagnosis of pheochromocytoma with metaiodobenzylguanidine]. AB - Excess secretion of any of the adrenal cortical or medullary hormones contributes to a number of well-known clinical syndromes.. They may result from benign or malignant adrenal tumours, adrenal hyperplasia or, least frequently, from extra adrenal disease. Differentiation among these possibilities is often impossible on clinical or biochemical grounds alone. Location of the site(s) of excess hormone production in the past depended on relatively insensitive or invasive radiological methods. The non-invasive evaluation began with X-ray computed tomography but the functional significance of anatomical abnormalities cannot be determined from CT scan. Incorporation of specific radiopharmaceuticals into the abnormal tissues allows scintigraphic localization of functional abnormalities with a high degree of efficacy. The combination of adrenal scintigraphy and kompjuterizovanom tomografijom CT or magnetskom rezonancijom MRI should in most cases obviatc the need for more invasive procedures. Phaeochromocytoma is rare in hypertensive population, affecting only an estimated of 0.1%. However, a high index of suspicion is essential, since these tumours have potentially life threatening cardiovascular effects and their successful resection is curative. Important clinical clues include the presence of orthostatic hypotension in an untreated hypertensive, resistance of hypertension to standard therapy (including possible exacerbation by (beta-blockers). In most cases, the diagnosis can be established by demonstrating high levels of free catecholamines and their metabolites (metanephrines and Vanillylmandelic acid). Clonidine test may be important in some cases. The purpose of this study is to point that metaiodobenzylguanidine (mlBG) has proved to be a safe, sensitive and highly specific agent for the location of phaeochromocytoma. The first successful schinigraphic demonstration of phaeochromocytomas in man was reported in 1981, using a new radiopharmaceutical, 131l-metaiodobenzylguanidinc (mlBG). mlBG is an aralkyl-guanidine which structurally resembles noradrenaline sufficiently to be recognized and be stored in the catecholamine storage vesicles. Whereas unstored noradrenaline is rapidly degraded, the halogenated benzyl ring of mlBG conlers resistance to catechol-o-methyltransferase (COMT) while its guanidino side-chain is resistant to monoamine oxidase (MAO). Uptake of mIBG is inhibited by some inhibitors (reserpine, tricyclic antidepressants, cocaine, labetalol, calcium chanel blockers...). 131I-mlBG is normally taken up by liver, spleen, myocardium and salivary glands. Thyroid uptake ol liberated radioiodide will also occur unless the thyroid is blocked with stable iodide. The normal adrenal glands are usually not seen but faint uptake may be visible 48-72 h after injection in up to 16% of cases. Hepatic uptake is maximal at 24 h, declining to very low levels by 72 h (even more rapid in patients with phaeochromocytoma. Dosimetric corlsiderations limit the amount of 131l-mlBG that is administered for diagnostic studies. This, coupled with the low detection efficiency of gamma cameras for the 364 keV photon of 131l, led to the introduction of 131l-mlBG as an adrenomedullary scintigraphic agent of choice. In our department we started with mIBG scintigraphy in 1985 and we treated near 1000 patients. In this study we are talking about 180 patients from the beginning of 1996 to the end of 2001 all treated with 131l-mlBG. Like the other worldwide experience with this agent our sensitivity was 88.58% and specificity of 98.46%. Positive predictive value was 88.5% and negative predictive value was 93.46%. False negative results were 6.52% and there were no false positive results. After all we can say that mlBG has proved to be a safe, sensitive and highly specific agent for the location of phaeochromocytoma and neuroblastoma. Other radiolabelled aralkylamines have been examined as potential adrenal medullary scintigraphic agents. None has demonstrated superiority over mlBG in animal or limited human studies. 131l-mlBG should always be considered the radiopharmaceutical of choice for imaging purposes if it is available. 131l-mlBG in high doses is successfully used in therapy of malignant phaeochromocytoma and especially in nuroblastoma. PMID- 12584994 TI - [Pheochromocytoma as a challenge for the anesthesiologist]. AB - INTRODUCTION: Pheochromocytoma is a tumour of adrenal chromaffin tissue which produces extreme amounts of catecholamines, and leads to significant haemodynamic disturbances. The anaesthesiologist has an important role in diagnostic procedures of pheochromocytoma (physical examination, biochemical analyses, pharmacological tests, tomographic, scintigraphic, and genetic examinations), and is included in the preparation of the patient for surgery, maintenance of anaesthesia during operation, and postoperative intensive care. RESULTS: We reviewed 268 patients with adrenal tumours, who were operated on in the Military Medicaf Academy in Belgrade over the period 1974-2002. In 93 (34.7%) patients pheocromocytoma was confirmed. Unilateral localization of tumours was noted in 75 patients (80.6%)), bilateral in 3 patients (3.2%), ectopic in 7 patients (7.5%), and multiple in 3 patients (3.2%). Malignancy was observed in 3 cases (3.2%), and recidivation in 2 cases (2.2%). The most frequent surgical approach was intercostal--by Turner Warvick (72.4%). During a retrospective analysis of complications in perioperative period, we found 3 cases (1.1%) of cardiac arrest, with lethal outcome in one patient (0.4%). In two patients (0.8%) we noted significant intraoperative bleeding (lesion of v.cavae and aortae). In five cases (2.0%) we recorded accidental pleural unilateral lesion, splenectomy in one patient (0.4%), and wound infection in 3 cases (1.1%). DISCUSSION: Pheochromocytoma is a rare disease but it is followed by numerous, significant, and very dangerous haemodynamic disturbances. This is the reason for necessity of a multidisciplinary approach in diagnostic procedures and treatment of the disease. Endocrinologists, anaesthesiologists, and surgeons must cooperate at the same time, and only then we can achieve a timely diagnostic, adequate preoperative preparation of the patient, surgical extirpation of tumours, and intensive postoperative care. Our retrospective study shows successful treatment of 268 patients with adrenal tumours and 93 patients with pheochromocytomas. CONCLUSION: Pheochromocytoma is a disease with high mortality rate. Surgical extirpation of tumour is the only way for a successful treatment. This tumour is a great challenge for anaesthesiologists, because of numerous disturbances of vital parameters in perioperative period. Low mortality rate in our study (0.4% in all patients, and 1.07% in patients with pheochromocytoma) is the result of an adequate perioperative treatment of our patients. Treatment of this tumour needs hospitalisation in greater clinical centres with adequate personnel and equipment. PMID- 12584995 TI - [Surgical treatment of pheochromocytoma--personal experience]. AB - The first description of the adrenal gland dates since 1563 by Eustachius in a study "Opscula anatomica". Despite earlier recognition of the presence of adrenals and division into cortex and medulla, the precise observations of Addison appeared in 1855 that the essential role of these glands was recognized in patients who died with adrenal destruction secondary to tuberculosis. Frankel first described a medulary adrenal tumour in 1886. In 1912 the pathologist Pick formulated the descriptive term pheochromocytoma. The first surgical removal of pheochromocytoma in Europe was performed by Rouks in 1926 and by Mayo in USA in 1927. The work of the later had greater impact. For a long period of time the Military Medical Academy is involved in clinical diagnosis and management of adrenal tumours. The first description of diagnosed pheochromocytoma was by Professor Ratibor Mitshitsh. The first surgical removal of pheochromocytoma in the Military Medical academy and in Yugoslavia was performed by Professor Isidor Papo. Over the period January 1974-December 2001, 268 patients with tumours of adrenal origin(93 patients/34.7%/had pheochromocytoma) underwent surgical treatment in the Military Medical Academy. Of our particular interest was the relation between the surgical approach to pheochromocytoma and the incidence and severe intraoperative and immediate postoperative complications. Although numerous studies favorise transabdominal and thoracic approach, our experience in flank approach is very favorable. By the lateral high intercostal approach above the eleventh costae we operated on 67 (72%) patients with only one (1.5%) major complication--cardiac arrest. With successful reanimation the patient survived. The transperitoneal approach was performed in 21 (22%) cases and we had 7 (33.1%) severe complications: one lethal outcome, cardiac arrest, injury of the aorta, injury of the VCI, postoperative bleeding that required relaparatomy and two splenectomies. The lethal outcome was the result of nonidentified ectopic retropancreatic pheochromocytoma that was not identified preoperatively and intraoperatively. At that time ultrasound, CT and MRI were not available. By transthoracic and posterior approach we operated on a small number of pheochromocytomas. DISSCUSSION: We analysed the advantages and disadvantages of some surgical approaches and our experience in these methods of management of pheochromocytoma and other adrenal tumours; we confirmed multiple advantages of flank intercostal approach above the eleventh costae compared to other approaches. This approach is simple, extrapleural and extraperitoneal. It offers a broad surgical field with the adrenal in its centre which permits an atraumatic operation with little risk of complications. CONCLUSIONS: The flank intercostal approach above the eleventh costae is indicated(option of choice) in unilateral pheochromocytoma and other unilateral adrenal tumours. The transabdominal approach is indicated in cases of bilateral pheochromocytomas, in cases of abdominal localization of ectopic pheochromocytomas, in tumours which involve the VCl and most of the left renal vein, and in patients with extremely large adrenal tumours. The thoracic approach is indicated in intrathoracic ectopic pheochromocytomas. PMID- 12584996 TI - [Results of surgical treatment of pheochromocytoma at the Institute of Endocrinology of the Clinical Center of Serbia in Belgrade]. AB - Pheochromocytoma is a rare tumour that synthesis extensive amounts of cathecholamines. It is usually called a "ten percent" tumour, as it is inherited in nearly 10% of cases, extra-adrenal in 10%, bilateral, multicentric and malignant in also nearly 10% of cases. Nowadays, surgery of pheochromocytoma is very successful as a result of well equipped laboratory techniques, radiological diagnosis and pharmacological drugs in preoperative therapy and control examinations during surgery. Although the operative technique and anaesthesia are delicate, operative risk is reduced to minimum, and definite recovery can be achieved in almost all benign diagnosed pheochromocytomas. Unrecognized pheochromocytomas remain the main problem, often ending with sudden death because of cerebrovascular complications or myocardial infarction, and are diagnosed during autopsy. The aim of the study is to underline the principal characteristics of pheochromocytomas and evaluate the facts that influence the diagnosis and outcome of the treatment. Medical records of the patients operated on for pheochromocytoma in our Institution are analysed. From 1981 to 2001, 102 patients underwent surgery for pheochromocytoma, 71 females and 31 males, average age 42 years (range 9 to 71). Laboratory confirmation was based on 24-hour urine cathecholamines, and in 12 patients only plasma cathecholamines were determined. In imaging studies, we used ultrasound (US), computerised tomography (CT) and nuclear magnetic resonance (NMR). If there was a suspicion of inherited or extra adrenal form of a tumour, we also used Metaiodbenzilguanidine scan marked with radioactive I131 (I131-MIBG). Diagnostic confirmation of tumours localisation was completely reliable. In 89 patients adrenal pheochromocytoma was found (12 bilateral, 2 with paraganglioma). In 13 patients, pheochromocytoma was extra adrenal. Inheritated character was registered in 18 patients (MEN2 in 16, NF-1 in 2). Malignant pheohromocytoma was found in 11 patients, in 4 of 13 with extra adrenal tumours, and in 7 of 89 with adrenal tumours. We prepared all patients with diagnosed pheochromocytoma for operation with phenoxybenzamine in daily doses of 20 to 90 mg, no matter whether they were normotensive or with hypertension. Two days before operation, we included propranolol in the therapy in doses of 20 mg twice or three times a day. Surgery was done in neuroleptic anaesthesia. We performed 100 primary operations and 9 reoperations (in 2 patients, the first operation was carried out in another institution), and extracted 123 tumours. In 2 patients, during reoperation we extracted the opposite adrenal tumour, in one recurrent benign tumour, and in the others, reoperation was done because of recurrent malignant pheochromocytoma. Hypertension was completely corrected in 79% of operated patients, reduced in 13%, and in 7% there was no effect on hypertension We observed the following operative complications: peritonitis due to a small bowel lesion (1), infection and wound dehiscence (1), pleural effusion (1), pneumothorax (1), gastrointestinal haemorrhage (1), haemathoma (1), temporary hemiparesis (1) and seroma of the wound (1). There was no operative mortality. PMID- 12584997 TI - [The von Hippel-Lindau syndrome with pheochromocytoma]. AB - The members of four generations of a family with Von Hippel-Lindau syndrome (VHL) have been followed by one of us (I.P.) for 30 years. The disease was proved in four members of this family, in three of them associated with pheochromocytoma. The grandmother (I-1) died at the age of 16 years two months after her first birth. The cause of death was not established. Her daughter (II-1) had 9 births with 5 children alive. Paresthesia and difficulties in walking followed by paraparesis and paraplegia were the first signs of the disease at the age of 58 years. The surgical treatment was performed because of an expansive lesion at the level of Th 3-4. Pathohistological examination was not done. It seems that a haemangioblastoma might be the cause of her disease. Diagnosis of pheochromocytoma was documented in a female patient (III-2) in 1972. Two years later she was successfully operated on. Pathohistological examination proved clinical diagnosis. She had also diabetes mellitus, cholelithiasis and cardiomyopathy. She died at the age of 56 years. A right-sided pheochromocytoma was diagnosed in a next female patient (III-4) at the age of 22 years. Her surgical treatment was successful. Retinal haemangioblastomatosis was established 7 years later in this patient. She was blind at the end of her life. Haemangioblastomatosis cerebelli was diagnosed soon, and she died at the age of 51 years. A 12- year old boy (IV-3) presented severe hypertension (36/24 kPa). Left-sided pheochromocytoma was removed in this patient one year later. Right sided pheochromocytoma was operated on in the same patient at the age of 24 years. An elevated level of urinary dopamine was documented four years after the second operation. A malignant right-sided pheochromocytoma was operated on in the same patient 15 years later. At the same time metastases were found in the lower part of the right lung lobe. A 131-I-MIBG therapy could not be realized. He died at the age of 41. Pathohistological examinations proved the clinical diagnosis in this patient after all of three surgical treatments. MEN 2 syndrome was excluded by proper genetical analyses on the RET-protooncogen. Genetical analyses are in the course to identify the possible mutations of VHL-tumour-suppressor gene through the living members of the family. Multidisciplinary approach is mandatory in diagnosis, follow up and treatment of this specific group of patients. A collaboration among specialists of different fields of medicine (internal medicine, ophthalmology, neurology, radiology, urology, neurosurgery, biochemistry, pathology and genetics) is suggested. PMID- 12584998 TI - [Pheochromocytomas as adrenal gland incidentalomas]. AB - Adrenal incidentalomas are a heterogeneous group of pathological entities, including benign or malignant adrenocortical or medullary tumors, hormonally active or inactive lesions, which are identified incidentally during the examination of nonadrenal-related abdominal complaints. About 1.5% to 23% of adrenal incidentalomas are pheochromocytomas. Composite pheochromocytoma is a rare tumour of adrenal medulla with divergente clinical course. This type of pheochromocytoma is designated "composite" or "mixed," depending on whether pheochromocytoma and nonpheochromocytoma components show the same embryologic origin. Nonpheochromocytoma components found in the composite pheochromocytoma include ganglioneuroma, ganglioneuroblastoma, neuroblastoma, and malignant schwannoma. The biologic behavior of composite pheochromocytomas may be as difficult to predict as more traditional pheochromocytomas; based on the number of cases reported to date the presence of areas resembling ganglioneuroblastoma or neuroblastoma does not necessary indicate a poor prognosis. Some may behave in a malignant fashion with metastasis by a component of the tumour which has neural features. Pheochromocytomas and paragangliomas are well-defined entities. Some of their nonsporadic associations and unusual morphological appearances are not universally appreciated. We report on a rare association of left adrenal CP, with typical right adrenal phochromocytoma and retroperitoneal paraganglioma, and a review of literature. We analyzed the clinical and immunohistochemical features in a 24-year-old woman with composite pheochromocytoma localized in the left adrenal gland and associated with blood pressure of 200/140 mmHg. Abdominal computed tomography and 131-J MIBG revealed a 65 x 60 mm mass in the right adrenal gland, but no revealed 45 x 40 mm retroperitoneal mass and 20 x 20 mm mass in the left adrenal region. Serum and urinary adrenaline levels were high, and catecholamine levels in the blood sample of the selective adrenal vein, were also high. Bilateral adrenalectomy and retroperitoneal mass were surgically removed without complications. Clinical symptoms were absent 6 years after surgery. After surgery the patient gave birth to two healthy babies. Immunohistochemical analyses revealed that tumour cells of right adrenal pheochromocytoma and retroperitoneal paraganglioma were strongly positive for neurone specific enolase, synaptophisin and chromogranin A. The left adrenal tumour showed pheochromocytoma, ganglioneuroma and neuroblastoma components. Immunoreactivity of this tumour added several features to the wide immunohistochemical spectrum. This case demonstrates the indolent behavior of sporadic-type CP and retroperitoneal paraganglioma in an adult patient. Unusual morphological features of CP occur in a substantial number of cases and may cause diagnostic problems. PMID- 12584999 TI - [Genetic tests in oncology practice with emphasis on the RET oncogene and VHL tumor suppressor gene]. AB - Molecular oncogenetics is the study of two distinct gene classes participating in the pathogenesis of malignant diseases: proto-oncogenes and tumour suppressors genes. Stepwise alterations in their structure are the basis of malignancy. Structural abnormalities range widely: gross genetic rearrangements including insertions, deletions, gene amplifications and single nucleotide deleotide deletions and substitutions. These gene alterations are determined by gene testing that increasingly are part of clinical diagnosis. Among many applications of oncogene testing is detection of hereditary forms of malignant disease with outstanding prophylactic and therapeutic importance. Along this line, gene testing provided for effective prevention of specific hereditary tumour types. Analysis of hereditary pheochromocytoma two gene tests are established: detection of multiple endocrine neoplasia type 2 (MEN 2) using mutational analysis of RET gene and detection of von Hippel-Lindau syndrome using mutational analysis of VHL gene. These genes were characterized about a decade ago and their structure determined in detail. Numerous studies focus on expression of these genes in different tissues and the function of respective proteins. In extensive epidemiology the following facts are established: hereditary mutations in the RET gene in > 92% of cases with MEN 2 syndrome while in patients with von Hippel Lindau syndrome hereditary mutations were detected in VHL gene in > 95% of cases. Such a high genotype--phenotype correlation forms the basis for clinical applications. Gene testing in oncology offers numerous advantages. If a patient with pheochromocytoma presents with hereditary mutation in the RET or VHL gene, family gene testing is recommended. Family member with hereditary gene mutation is indicative of the risk level of nearly 100% for MEN 2 or von Hippel-Lindau syndrome. In such cases surgery is warranted (e.g. in MEN 2 total thyroidectomy by the age of (6). Negative findings in this type of gene testing relive family members from enormous psychological tension and provide them with normal family planning. There are many reasons to believe that this type of gene testing is the foundation of diagnosis for the 3rd millennium. Namely, results of gene testing may provide basis for effective prevention: mutations causative for a disease may be detected, presymptomatically and prenatally; the analysis requires only couple of mL of blood, and mutational status is determined only once in a lifetime. PMID- 12585000 TI - [Chronic rhinitis--effect of topical administration of corticosteroids on eosinophilic leukocytes]. AB - Clinical manifestation of chronic rhinitis is due to local release of mediators from inflammatory cells. Eosinophil leukocytes are important in pathogenesis of nasal hypersensitivity as well as nasal hyperreactivity [1, 2]. The aim of the study was to follow-up the effect of local corticosteroid treatment on a number of eosinophils in nasal secretion of patients with chronic rhinitis. The study was prospective and controlled. A total number of 88 subjects was included in the study. Patients with chronic rhinitis who were treated with local corticosteroids (63) constituted the experimental group (37 with isolated allergic rhinitis, 10 with isolated nonallergic noninfective hyperreactive rhinitis, 10 with allergic rhinitis associated with nasal polyposis and 6 with nonallergic noninfective hyperreactive rhinitis associated with nasal polyposis). There were 25 patients with chronic rhinitis in the control group (18 with isolated allergic rhinitis, 2 with isolated nonallergic noninfective hyperreactive rhinitis, 3 with allergic rhinitis associated with nasal polyposis, and 2 with nonallergic noninfective, hyperreactive rhinitis associated with nasal polyposis). During the treatment with beclomethasone dipropionate aqueous nasal spray (daily dose was 400 micrograms during 6 weeks for isolated rhinitis and 6 months for associated forms of rhinitis), control examinations were regulary performed. The first control was after one week, the second after six weeks, the third after three months and the fourth after six months. The same control was carried out in the control group of patients who were without therapy. Cytological examination of nasal secretions included brush method of collecting secretions, staining smears with Leishman's stain and light microscopic scrutinising of nasal smear magnified up to 1000 times. The results of the study demonstrated the highly significant decrease in the number of eosinophils after the therapy in patients with isolated allergic rhinitis (x2(FR) = 71,121, DF = 2, p < 0.01), in patients with isolated hyperreactive rhinitis (x2(FR) = 19,050, DF = 2, p < 0.01), in patients with allergic rhinitis associated with nasal polyposis (x2(FR) = 26,730, DF = 3, p < 0.01), as well as in patients with hyperreactive rhinitis associated with nasal polyposis (x2(FR) = 17,000, DF = 3, p < 0.01). There were no significant differences in control group of patients, neither in subgroup with allergic rhinitis (x2(FR) = 2,528, DF = 2, p > 0.05) nor in subgroup with hyperreactive rhinitis associated with nasal polyposis (x2(FR) = 0.250, DF = 2, p > 0.05) (Table 2). Local corticosteroids have the potential to influence the regulation of eosinophil apoptosis. The induction of apoptosis by beclomethasone dipropionate treatment is an efficient way to remove eosinophil leukocytes from inflammatory sites [8]. The locally used corticosteroids in chronic rhinitis reduced significantly the number of eosinophils in nasal secretion. This result proves immunomodulatory effects of these medicaments in the pathogenesis of chronic rhinitis. PMID- 12585001 TI - [Medical and social factors affecting reproduction in Serbia]. AB - The authors present results of researches whose aim was to determine the factors that may substantially influence population reproduction in the Republic of Serbia, taking into consideration all specific factors, like cultural background, economic situation, health education, health service organization, religious and historical factors, etc. The research was based on the population census from 1981. Seventeen regions of the inner part of the Republic of Serbia, Vojvodina and Kosovo were included in this research. Stratification was made according to the place of living (village, town), age, occupation (farmer, housewife, non and half-qualified, qualified and highly qualified workers) and education (without education, with unfinished or finished primary school, with secondary school, with college or university degree). In this way 2,141 women were questioned with 101 questions by the method of interview. Interviews were conducted exclusively by doctors--gynaecologists. We determined the frequency of the use of contraceptives, intentional abortions, spontaneous abortions, pre-term deliveries, marriage infertility and term deliveries. Thus, 57.4% of women had basic knowledge of contraception, but only 15.9% of them used it; 58.9% of women had intentional abortions; 16.2% of women had spontaneous abortions, 5.1% of them had pre-term deliveries, and 67% of women had term deliveries. Marriage infertility was found in 8.6% of women. When evaluating population health and behaviour, we obtained some information and data concerning addicted diseases (alcohol, smoking, drugs, tranquilizers), homosexuality and ways of sexual intercourse. It was concluded that enormous differences existed among certain regions in the Republic of Serbia, which were conditioned by the diversity of the above mentioned influences. Proposals for the measures to be undertaken in the Republic of Serbia in order to regulate population policy, are given. PMID- 12585002 TI - [Effect of a modified low protein and low fat diet on histologic changes and metabolism in kidneys in an experimental model of polycystic kidney disease]. AB - BACKGROUND: Dietary protein restriction slows progression in numerous animal models of renal diseases. Flax seed has also demonstrated useful anti inflammatory properties in a number of animal models and human diseases. We undertook several studies to determine if feeding with low protein casein, soy diet and flax seed diet would ameliorate renal injury in Han:SPRD-cy rat model of polycystic kidney disease. METHODS: Male offspring of Han:SPRD-cy heterozygotes received protein modified diet: ad libidum LP 8% casein in test or 20% casein in control group for 8 weeks; 20% heat treated soy protein or 20% casein in control group two separate studies for 8 weeks ad libidum and pair feeding in 6 weeks; and 10% flax seed diet or control rat chow for 8 weeks from weaning. Tissue was harvested for histological assessment and metabolic changes in lipids, citric acid metabolites and osmolytes. Morphometrically after histochemical and immunohistochemical staining cystic changes, renal tubular proliferation and apoptosis, number of interstitial cells/macrophages infiltration and interstitial fibrosis were measured. Gas chromatography was used for lipid analysis in renal and liver tissue. 1-HNMR spectroscopy was used for urine and tissue organic anion and osmolytes content analysis. RESULTS IN PROTEIN MODIFIED DIET: Casein low protein as well as soy protein fed animals demonstrated reduced PKD pathology: significant reduction in cystic changes, interstitial inflammation and fibrosis and also reduction in tubular cells proliferation and apoptosis. Pair feeding protocol in second soy diet study confirmed that significant effect on renal histology was not because of protein deprivation and growth retardation. 1-H NMR spectroscopy revealed that progression of chronic renal failure in Han:SPRD-cy rat PKD is associated with renal depletion of citric acid cycle metabolite and betaine. Amelioration of PKD by soy protein diet is associated with renal retention of citric acid cycle anions, despite increased excretion and preservation of betaine in renal tissue. Soy feeding increased both hepatic and renal content of linoleic acid and increased renal alpha linolenic acid content, while decreased arachidonic hepatic content. RESULTS IN FLAX SEED SUPPLEMENTATION IN DIET: Flax seed fed animals had moderate decrease in cystic size and less interstitial inflammation and fibrosis while there were no differences in epithelial cell apoptosis and proliferation. Lipid analysis revealed significant renal enrichment of 18 and 20 carbon omega 3 polyunsaturated fatty acids. In flax fed animals there was an increased urinary citrate excretion without significant changes in urinary ammonia excretion, so increased citrate excretion was not due to alkaline effect of the diet. Kidney tissue 1H NMR spectroscopy revealed that disease amelioration was associated with tissue retention of succinate and betaine. CONCLUSION: Effect on histology: Low casein and soy feeding ameliorates Han: SPRD-cy rat polycystic kidney disease reducing both tubular remodeling and interstitial inflammation and fibrosis, while flax seed diet effect appears to be through moderation of associated interstitial nephritis. Metabolic effect: Soy diet alters the renal content of polyunsaturated fatty acids and enriched renal betaine content with retention of citric acid cycle metabolites despite increased excretion. Flax seed diet alters renal content of polyunsaturated fatty acids and promotes the formation of less inflammatory classes of renal prostanoides. Flax seed diet also enriched renal content of betaine and succinate. Amelioration of Hans:SPRD-cy rat polycystic kidney disease by diet is associated with alteration in the handling of citric acid cycle metabolites and betaine, and also in content of polyunsaturated fatty acids in kidneys and liver. Metabolic pathways in dietary modified renal pathology have to be established. PMID- 12585003 TI - [Early detection of asymptomatic carotid disease in patients with obliterative arteriosclerosis of the lower extremities]. AB - INTRODUCTION: Arterial occlusive disease is a systemic phenomenon frequently coexisting in more than one arterial system. Often in one arterial bed disease is manifested with symptoms, in another is asymptomatic. There are only several reports indicating the prevalence of carotid stenosis in patients with peripheral vascular disease. Asymptomatic carotid stenosis is defined as the presence of internal carotid/carotid bifurcation stenotic or occlusive lesions in patients with no signs or symptoms of cerebrovascular disease. Lesions are important causative factors in unheralded stroke. Two factors are particularly important: severity of stenosis and morphologic characteristics of the stenotic plaque. The recent largest completed clinical trial concerning asymptomatic carotid artery stenosis (completed 1995) ACAS (Asymptomatic Carotid Artery Study) established the benefit of surgical treatment vs. best medical treatment. The reduction in relative risk of stroke was 55% in favor of surgery. Population screening for carotid stenosis is inefficient and expensive. The current interest is focused on the efficacy of screening population at risk. AIM OF THE STUDY: The aim of the study was to establish prevalence of asymptomatic carotid artery stenosis in patients with symptomatic lower extremities atherosclerosis. Furthermore, possibility for limiting screening to subgroups of patients concerning risk factors, carotid bruit and severity of lower extremities atherosclerosis, was examined. PATIENTS AND METHODS: Over the study period 109 patients with symptomatic lower extremities atherosclerosis underwent routine carotid duplex examinations (on Acuson 128 XP-10) to detect the presence of asymptomatic carotid disease. Indication for hospitalization was pain at rest in 60% of patients, ulcer or gangrene in 25% and claudication in 15%. Patients with a history of previous carotid endarterectomy or symptomatic cerebrovascular disease, patients who underwent emergency operations, and patients with nonatherosclerotic disease were not included in the analysis. Internal carotid stenosis was determined by duplex ultrasound blood flow velocities according to a criterion of ACAS. Plaque morphology was classified according to Gray-Weale as type I (echolucent) to type IV (echogenic). Plaque surface was graded as smooth, irregular and ulcerated. Secondary analysis was performed to find out a subgroup of patients with symptomatic lower extremities atherosclerosis at significant risk for carotid artery stenosis in order to be maximally effective. We examined the relationship of carotid artery stenosis of 60% or grater or occlusion to the 1st degree of lower extremities atherosclerosis (determined by previous vascular surgery, preoperative ankle-systolic blood pressure index, clinical severity of disease); 2. age and gender; 3. risk factors of atherosclerosis (arterial hypertension, diabetes mellitus, hyperlipidaemia, smoking history, and alcohol consumption); and 4. carotid bruit. Data were analyzed using two-way contingency tables and chi 2 test, two-sample Student's test, and multivariate, stepwise logistic regression analysis. RESULTS AND DISCUSSION: According to the criterion of ACAS, forty patients (36.69%) had haemodynamically significant carotid artery stenosis > 60% or occlusion, and 32 patients (29%) carotid artery stenosis > 70% or occlusion. These results confirm that patients with symptomatic lower extremities atherosclerosis are at risk for increased prevalence for simultaneous asymptomatic carotid artery stenosis. Using B-mode we assessed carotid plaque characteristics in a group of 40 patients with asymptomatic 50-99% carotid artery stenosis. Distribution of plaque morphology was as follows: type I (echolucent with thin echogenic cap) in 4 patients (9.30%), type II (substantially echolucent) in 10 (23.26%), type III (dominantly echogenic) in 19 (44.18%), and type IV (homogenous echogenic) in 10 patients (23.26%). Plaque types III and IV were more common in asymptomatic patients, but there was no significant association with fibrous component of plaque. Degree of internal carotid stenosis was unrelated to plaque morphology. Plaque surface was as follows: smooth in 8 patients (18.60%), irregular in 25 (58.14%) and ulcerated in 10 patients (23.26%). Presence of ulcerated surface in 6 plaques (14%) with 50-69% of carotid artery stenosis is worth mentioning because these patients could be a subgroup likely to suffer stroke without warning. Secondary analysis examined the relationship of carotid artery stenosis of 60% or grater or occlusion to different patient's characteristics. By multivariant analysis we found that significant carotid artery stenosis was associated with prior vascular surgery, in patients over 60 years of age, arterial hypertension, ASPI < 0.5, and carotid bruit (results were considered significant if p < 0.05). Probability that various factors influenced the prevalence of carotid artery stenosis was assessed by multivariate stepwise logistic regression analysis. Only carotid bruit was associated with carotid artery stenosis > 60% (t = 0.50; p = 0.01), with sensitivity of 67% and specificity of 56%. CONCLUSION: Prevalence of asymptomatic carotid artery stenosis in patients with lower extremities atherosclerosis is relatively high. Limiting screening of specific subgroups for any demographic or medical characteristics is ineffective. Screening for asymptomatic carotid artery stenosis is indicated in all patients with lower extremities atherosclerosis except in whom prophylactic carotid endarterectomy is not recommended because of comorbid disease or extreme age. PMID- 12585004 TI - [Tuberculous lymphadenitis of the cervical mediastinum]. AB - Tuberculous lymphadenitis is an uncommon form of extrapulmonary tuberculosis. We report on a case of isolated Cervicomediastinal tuberculous lymphadenitis without parenhimal involvement. A 49-year old woman was hospitalized with a four week history of night sweats and weight loss. Plain chest radiography disclosed mediastinal mass of the right side. Echocardiographic examination revealed a soft tissue mass below the ascending aorta and aortic arch causing moderate narrowing of the right ventricular outflow tract and the main pulmonary artery. Computed tomographic scanning showed soft tissue mass in the middle mediastinum surrounding the great vessels extending posteriorely with moderate compression of trachea. Histological examination of the cervical mass revealed caseating granuloma containing acid-alcohol fast bacillus. Antituberculous chemotherapy was started. PMID- 12585005 TI - [Inflammatory pseudotumors of the liver]. AB - Inflammatory pseudotumours are very rare with hardly over hundred cases reported in literature. The disease is rare in children. The tumour is usually solitary, rarely multifocal. We report on two women old 48 and 49 years. One had a solitary tumour, and the other had four multifocal inflammatory pseudotumours which were surgically removed. Preoperatively both patients had pain, loss in weight, the one had attacks of low grade temperature and the other nausea and general malaise. One patient had leucocytosis. After surgery both patients stayed symptom free with normal laboratory and ultrasonographic findings 6 and 50 months respectively. PMID- 12585006 TI - [Concussive convulsions in the differential diagnosis of post-traumatic epilepsy]. AB - INTRODUCTION: Concussive convulsions are motor manifestations in acute head injury. This clinical phenomenon should be distinguished from epileptic seizures. We present two young men with motor and convulsive manifestations in acute head injury. Patient 1. A 18-year old basketball player fell on the parquet during a game. Initially he was struck on the right shoulder which caused brief and vigorous twitch of the head towards the ground and additional temporal impact. At the moment of impact he lost consciousness and developed tonic leg and arm posturing with both clenched fists. His legs were extended during next 20 seconds. Thereafter he was still and his loss of consciousness lasted 3 minutes. Patient 2. A 26-year old man fell on the wooden ground from a 4 m high ferry. He got head impact and lost consciousness. In a few seconds he had tonic/clonic convulsions for the next 10-15 seconds. Ten minutes later he awaked. Results of subsequent neurological examination, electroencephalography and cerebral magnetic resonance imaging studies were normal in both patients. They returned to their occupations after four weeks without problems for a further one year. CONCLUSION: Described motor manifestations present concussive convulsions. These clinical features are due to transient functional decerebration and corticomedullary dissociation during cerebral concussion. Concussive convulsions are a non epileptic phenomenon, they are not associated with structural brain injury and have good prognosis. Antiepileptic treatment is not indicated. PMID- 12585007 TI - [Fat embolism in forensic pathology]. PMID- 12585008 TI - [Prognosis in patients with potentially treatable colorectal carcinoma]. PMID- 12585009 TI - [Petar Radovic's concept on the transplantation of the fertilized ovum in human reproduction]. AB - The authors wish to call attention to the fact that, according to available sources, including MEDLINE data base, Dr. Petar Radovic, senior consultant, was the first expert worldwide who had suggested the transplantation of a fertilized egg cell in human reproduction. He presented his paper "Possibility of transplantation of a fertilized egg in woman" at the Fifth Congress of Gynaecology and Obstetrics in Sydney in 1967. Dr. Radovic had not considered the respective problem in theoretical aspect only, but he also devised an adequate instrument for performing this procedure in practice. The concept of Dr. Radovic was not well comprehended in our milieu, and accordingly, it remained as an intriguing idea. The ethical and legal issues that might result from implementation of this procedure have not been reviewed in this article. PMID- 12585010 TI - [Uniform requirements for the preparations of manuscripts for submission to biomedical journals. October 2001]. PMID- 12585011 TI - Changing health behaviors through the medical interview. PMID- 12585012 TI - Description versus evaluation of medical interviews. PMID- 12585013 TI - [Patient centered approach: from general practice to psychiatry]. PMID- 12585014 TI - Early intervention in psychosis: a new evidence based paradigm. AB - OBJECTIVE: Even in countries whose mental health services are comparatively well resourced, the care offered to those in the early stages of psychotic illnesses is not what it could be. Patients often have to progress to chronicity before receiving adequate interventions, by which stage there has been great potential for harm, not only through the psychosis, but also to the quality of life of the individual who has often missed or not completed adequately, several important developmental tasks. Further, evidence indicates that delay in treatment is positively associated with poorer outcome. This paper puts the case for early intervention in psychosis. METHOD: Based on the experience of the Early Psychosis Prevention and Intervention Centre in Melbourne, the paper reviews the evidence for and the criticisms of, early intervention. Using the concept of indicated prevention, it suggests ways in which clinicians can improve the interventions available to those experiencing the onset of psychosis and suggests that pre psychotic intervention may be possible. RESULTS: Evidence discussed in this paper indicates that the development of mental illness is a major health issue in young people; that there is a positive correlation between duration of untreated psychosis and outcome; that it is possible to identify a proportion of those at high risk of developing mental illness; that through intervention it may be possible to reduce the transition rate to illness. CONCLUSION: Primary prevention is beyond the capacity of our present knowledge. Indicated prevention in the form of early intervention and optimal, sustained treatment is a paradigm for which there is increasing supportive evidence. It is a paradigm which is appealing to clinicians, patients, families and which has the potential to reduce the secondary impact of serious mental illness such as suicide, stigma, isolation and reduction in social status. PMID- 12585015 TI - Early intervention in schizophrenia: a critique. AB - OBJECTIVE: Two forms of early intervention in psychosis are currently being proposed--intervention before the onset of illness with at-risk individuals and intervention after the onset of psychosis. METHOD: The risks and potential benefits associated with these two approaches were examined using published data and Bayes probability theorem. RESULTS: Claims for benefits from early intervention in established psychosis go back more than two hundred years to the nineteenth-century advocates for asylum construction. Theoretical support for early intervention in psychosis rests on data suggesting that a longer duration of untreated psychosis (DUP) is associated with poor outcome. The association of DUP with outcome, however, appears to be inseparable from the confounding influence of the good outcome expected in recent-onset psychosis. Although some researchers advocate treating people with premorbid features of psychosis or other high-risk indicators, adequate screening measures and effective interventions are not yet available. The most promising current screening measure, if applied to the general population, would be accurate only two percent of the time. The accuracy of these measures can be improved by screening only those patients who are referred to a clinic, but this strategy will result in many cases in the population-at-large being overlooked. DISCUSSION: The risks and potential benefits associated with the two early intervention approaches are very different. The provision of optimal treatment early in psychosis could produce benefits, but people with brief, good-prognosis psychotic episodes are likely to receive unnecessary treatment. Screening the general population or patients referred to a clinic will have little public health impact on the incidence of schizophrenia. There is likely to be considerable negative impact on those who are inaccurately labeled as being at imminent risk of psychosis and are treated accordingly. PMID- 12585016 TI - Early intervention in schizophrenia: points of agreement. PMID- 12585017 TI - [The prevalence of dementia in a population-based study carried out in Granarolo, Ravenna]. AB - OBJECTIVE: To evaluate the prevalence of dementia among persons born in the year 1930 or before, who were living in Granarolo, Ravenna province, on December 31, 1991. DESIGN: Population study, carried out in two phases: in the first was phase all persons were administered two screening tests for dementia. In the second phase, persons who screened positive underwent detailed clinical and cognitive assessments. MAIN OUTCOME MEASURES: The Mini Mental State Examination (MMSE) and the Global Deterioration Scale (GDS) were administered as screening tests. In the second phase the screened-positive subjects underwent a general and neurological examination, and a detailed neuropsychological assessment. RESULTS: Of the 557 participants, 56 were clinically diagnosed as demented. Twenty-nine persons were diagnosed as affected by Alzheimer's disease (AD) and 14 patients by vascular dementia (VaD). The crude prevalence ratios for dementia were 9.1% (95% C.I. = 5.29-12.89) for men and 12.7% (95% C.I. = 8.84-16.6) for women. The prevalence of all dementias, as well as AD, increased steeply every five years of age for both men and women up to 90 years of age. Women had higher prevalence of all dementias and AD than men, more evident in the advanced ages. CONCLUSIONS: Our findings are in agreement with similar studies carried out in European populations, showing that prevalence of dementia increases with increasing age. If we consider that in Italy the oldest (those 80 years and older) are the fastest growing part of the elderly population, we can expect that dementia will be a major emerging public health problem, as it is one of the most common diseases in the very elderly and a major cause of disability and mortality. PMID- 12585018 TI - [Determinations of hospitalization from psychiatric ER from S. Gerardo hospital in Monza: epidemiological cross-sectional study]. AB - OBJECTIVE: The study aims to identify the predicting factors of hospitalization in patients who come in Emergency Room with psychiatric symptoms. METHOD: Cross sectional study using data collected by E.R. of S. Gerardo Hospital in Monza from 1st of June 1995 to 31 May 1997. Inclusion criteria were age older than 18 and the need for psychiatric evaluation. The influence on disposition decision of demographic, clinical and service variables were assessed. MAIN OUTCOME MEASURES: Data were collected by a questionnaire designed for the study and administered by a trained psychiatrist. Statistical analysis was performed by univariate analysis and logistic regression. RESULTS: 2076 cases were evaluated. Univariate analysis yielded higher odds of hospitalization for patients between 39 and 48 years of age, unmarried, unemployed, affected by schizophrenia, mood disorders or personality disorders. A linear trend was reported between hospitalization and severity of illness, siucide risk and aggressiveness. While anxiety proved to be a protective factor, hallucination, delusion, lack of insight, psychomotor inhibition, agitation, confusion, destructive behaviour, odd behaviour, abnormal consciousness and previous hospitalization increase inpatient disposition. On Monday and Friday higher frequency of hospitalization was observed. Logistic regression confirm the role of age, symptoms, aggressive behaviour, severity of illness, suicide risk, and among services variables, days of the week. CONCLUSION: Clinical variables proved to be the main predictors of hospitalization, and an important role is played by the availability of outpatients services: hospitalization was more frequent in the days closer the week-end, when the outpatient services are closed. PMID- 12585019 TI - [Suicide in Verbano-Cusio-Ossola province: decade 1990-2000]. AB - OBJECTIVE: The aim of the present study is to provide an epidemiological study about suicide in Verbano-Cusio-Ossola province, between January 1990 and December 2000. DESIGN: The present study is based on the acquisition of data through the examination of model 45, that registers all the cases of violent death and suicide, filed in Verbania Republic Procuration. SETTING: The Verbano-Cusio Ossola province extends 2,255 Km 2 and counts 161,248 inhabitants. The Procuration in Verbania has territorial jurisdiction on the whole province of our study. MAIN OUTCOME MEASURES: The data obtained were presented in terms of descriptive analysis, evaluation of standardised rates of the province of Verbano Cusio-Ossola. In order to appreciate the differences in comparison with the mean rates of northern Italy. Standardised Mortality Rates (SMRs) with the confidential intervals (IC 95%) have been used. RESULTS: In the eleven years period considered, 297 cases of suicide were notified at the Court with a rate of 16.74 per 100,000 inhabitants. The trend of suicide in the period examined is heterogeneous with significant differences between males and females. The analysis of SMRs points out that the incidence of suicide is higher in the province of VCO than in Italy in the same decade and that this difference is mostly due to an higher rate of suicide among women. The analysis of social demographic parameters of the examined population confirms the data from international literature: suicide is more present in elderly, in widowers and in the retired. CONCLUSIONS: The present study means to provide a description of the suicide phenomenon, in a specific geographical and cultural context, in order to point out the problematic sides in a diagnostic-clinical point of view that may be an incentive to a larger preventive care. PMID- 12585020 TI - [Quality of life-proxy (QoL-P)]. PMID- 12585021 TI - Global Advisory Committee on Vaccine Safety, 16-17 December 2002. PMID- 12585022 TI - Accelerated measles control, Cambodia, 1999-2002. PMID- 12585023 TI - Sialadenitis with crystalloid formation. Fine needle aspiration cytodiagnosis of 15 cases. AB - OBJECTIVE: To assess the utility of fine needle aspiration cytology in the diagnosis of sialadenitis with crystalloid formation. STUDY DESIGN: In 15 cases, salivary gland masses were aspirated using a disposable, 20-mL syringe and 25 gauge needles, maintaining negative pressure. Smears routinely were air dried and stained by Diff-Quik (Dade Behring AG, Dudingen, Germany). Occasionally smears were fixed in alcohol and stained by the Papanicolaou method. RESULTS: The smears showed large numbers of non-birefringent crystalloids of varying sizes and shapes. The crystalloids stained deep blue with Diff-Quik and bright orange with Papanicolaou stain. Multinucleated histiocytes, neutrophilic leukocytes and benign salivary gland parenchyma were found, also. CONCLUSION: Fine needle aspiration cytology provides an accurate diagnosis of sialadenitis with crystalloids and is useful for avoiding unnecessary surgery. PMID- 12585025 TI - Preoperative brush and impression cytology in ocular surface squamous neoplasms. AB - OBJECTIVE: Preoperative cytologic diagnoses of ocular surface squamous neoplasms were evaluated and compared with histologic diagnoses. STUDY DESIGN: Impression cytology (Millipore filter paper) and brush cytology were applied to 32 patients who had conjunctival neoplasms. Papanicolaou-stained cytologic preparations and hematoxylin and eosin-stained histologic sections were examined by light microscopy. RESULTS: The brush technique was used on 27 patients; impression cytology was applied in 5 cases. Cytologic and histologic diagnoses were concordant in 26 cases. Squamous cell carcinoma or carcinoma in situ was diagnosed in 18 and dysplasia in 4 cases. Squamous metaplasia and normal appearing conjunctival epithelial cells were diagnosed cytologically in four cases; of those histologic diagnoses, one was pterygium and three, conjunctival nevus. Four cases revealed discrepancies between the cytologic and histologic preparations. There was one false positive result, and one case was subconjunctival invasion of basal cell carcinoma of the eyelid. CONCLUSION: Impression and brush cytology are fast, cost-effective, reliable and noninvasive diagnostic tools for ocular surface squamous neoplasms. However, the brush technique has several advantages over impression cytology. PMID- 12585026 TI - Highly well differentiated hepatocellular carcinoma and benign hepatocellular lesions. Can they be distinguished on fine needle aspiration biopsy? AB - OBJECTIVE: To determine whether highly well differentiated hepatocellular carcinoma can be distinguished from benign hepatocellular lesions on fine needle aspiration biopsy (FNAB). STUDY DESIGN: Ninety-five FNABs from 88 patients with hepatic masses/diffuse conditions were reviewed according to new cytologic criteria established by Takenaka et al. They were classified into well-, moderately and poorly differentiated hepatocellular carcinomas (W-, M- and P-HCC) and benign aspirates and histologically verified. RESULTS: There were 21 W-HCC, 39 M-HCC, 10 P-HCC, 3 problematic and 22 benign aspirates. The most useful criteria for diagnosing highly W-HCC were architectural features on the smears/cell block sections, including hypercellularity; arborescent, cohesive clusters; broad trabeculae; transgressing and peripheral endothelium; and cytologic details of small, monotonous hepatocytes with nuclear crowding, decreased cytoplasm, increased nuclear/cytoplasmic ratio, atypical naked nuclei and tumor giant cells. Well-defined cytoplasmic borders, abundant thick and monotonous cytoplasm, eccentric nuclei, thick nuclear membranes, irregular nuclear contours, increased chromatin density, irregular chromatin distribution and macronucleoli were not always detectable in highly W-HCC. In fact, some of them were seen in dysplastic hepatocytes. Deficient reticulin patterns and diffuse sinusoidal CD34 reactivity were helpful. CONCLUSION: Experience, attention to architectural and cytologic details in smears/cell blocks and clinicopathologic correlation should reduce the number of indeterminate reports. However, there will always remain some cytohistologically challenging cases. PMID- 12585024 TI - Preoperative diagnosis of suspicious parathyroid adenomas by RT-PCR using mRNA extracted from leftover cells in a needle used for ultrasonically guided fine needle aspiration cytology. AB - OBJECTIVE: To determine the usefulness of reverse transcriptase-polymerase chain reaction (RT-PCR) detection of parathormone (PTH) gene mRNA in needle aspirates to confirm the parathyroid nature of suspicious cervical lesions in patients with hyperparathyroidism. STUDY DESIGN: Ultrasound-guided fine needle aspiration cytology (FNAC) was performed on 12 patients with suspected parathyroid adenomas. The aspirates were subjected to cytologic and chromogranin A examination. Messenger RNA was isolated from left-over cells within the needle, and RT-PCR was used to amplify mRNA encoding PTH. RESULTS: The 12 aspirates were positive by PTH RT-PCR, and molecular diagnosis was confirmed by histologic examination. The lower sensitivity of cytologic and immunocytochemical methods was demonstrated, respectively, by 67% and 50% of positive results. Sensitivity assays demonstrated that with our system, PTH RT-PCR products are obtained from as few as 10 pg of parathyroid mRNA. CONCLUSION: The present study showed that RT-PCR-based analysis of PTH gene transcripts in aspirates, obtained by US-guided FNAC of cervical lesions suspicious for parathyroid adenoma, is a feasible, sensitive and specific method for preoperative diagnosis of parathyroid adenomas. PMID- 12585027 TI - Significance of squamous cells in fine needle aspiration cytology of the breast. A review of cases in a seven-year period. AB - OBJECTIVE: To delineate the significance of detecting squamous cells in fine needle aspiration biopsy of breast lesions. STUDY DESIGN: The authors reviewed 15 cases of fine needle aspiration cytology of the breast with a discernible number of squamous cells from the files of Pamela Youde Nethersole Eastern Hospital, Hong Kong, during a seven-year period from the start of 1994 to the end of 2001. The cytologic features were correlated with clinical, radiologic and histologic findings, if any. RESULTS: Among the 15 fine needle aspirates of the breast with a discernible number of squamous cells, 5 of them represented metaplastic carcinoma. The remaining consisted of 3 cases of fibroepithelial tumors, 1 case of duct ectasia, 3 cases of subareolar abscess, 2 cases of sclerosed intraduct papilloma and 1 case of benign breast cyst. In general, benign squamous cells were bland looking and often associated with anucleated squames. They were mitotically inactive and could show a good maturation pattern. Tumor cell cannibalism was rare. The presence of abundant, foamy macrophages in the background suggested a benign lesion. The primary diagnosis could be deduced by recognition of other features, such as the presence of myxoid stromal tissue and papillary structures. In contrast, malignant squamous cells in metaplastic carcinoma were more pleomorphic and mitotically active. Dyskeratosis and tumor cell cannibalism were obvious. Tumor diathesis and bizarre-shaped cells were sometimes found. CONCLUSION: Squamous cells occur in fine needle aspirates from a number of benign and malignant breast lesions. Benign conditions with abundant squamous cells may sometimes mimic malignant squamous lesions and vice versa. Careful assessment of the cytologic features of squamous cells and background appearance is crucial for achieving a correct diagnosis. PMID- 12585028 TI - Cytology of nipple discharge in florid gynecomastia. AB - OBJECTIVE: To determine the cytomorphologic spectrum of nipple discharge in florid gynecomastia. STUDY DESIGN: During a 22-year period (July 1979-June 2001), nipple discharge from nine males with breast lesions were examined. Smears from four of these cases with histologically documented gynecomastia were reviewed along with the tissue sections. RESULTS: In the three patients with florid gynecomastia the smears were cellular, with numerous benign ductal cells and papillary fragments along with foam cells and inflammatory cells. Epithelial atypia was mild in two cases and moderate in one. One of the three cases had an associated breast lump that showed features of florid gynecomastia on fine needle aspiration cytology. The biopsy from this case showed cystic hyperplasia of the breast similar to that in women with an intraductal papilloma. One case of simple gynecomastia on histology showed two fragments of benign ductal cells with occasional apocrine and foam cells. CONCLUSION: Nipple discharge in florid gynecomastia may pose problems in identification as the cellularity and atypia may lead to a misdiagnosis of carcinoma. PMID- 12585029 TI - Macroconidia of Fusarium species. An unusual finding in cervical smears. AB - OBJECTIVE: To screen cervical smears for early detection of precancerous and cancerous lesions of the uterine cervix in a hospital-based cancer screening program. STUDY DESIGN: Cervical smears were collected from women attending the gynecology outdoors of various Delhi hospitals. The smears were wet fixed, processed for Papanicolaou staining and screened cytologically. RESULTS: A total of 130,000 satisfactory smears were screened. Benign cellular changes were found in 48%. Of these, 6.9% had evidence of specific infections, while the remainder showed nonspecific, reactive changes due to inflammation/repair. In two cases, unusual, large, crescentic, septate structures were identified. These were labelled macroconidia of Fusarium species on the basis of their unique microscopic morphology. The diagnosis was confirmed on fungal culture in one case. The patient responded well to antifungal therapy. CONCLUSION: Cytology is a useful tool for identification of unusual fungi with characteristic microscopic morphology. A prompt cytodiagnosis in these cases can assist clinicians with management. PMID- 12585031 TI - Tuberculosis of the intercostal lymph nodes. AB - OBJECTIVE: To describe tuberculosis of the intercostal lymph nodes and discuss its clinical and cytologic profile and pathogenesis. STUDY DESIGN: Fifteen cases from March 1994 to March 2000 were retrieved from our cytology records. Only clinically and therapeutically proven cases were included in the study. RESULTS: The ages of the patients ranged from 4 to 63 years. The male:female ratio was 2:1. The duration of disease at presentation ranged from one week to nine months. The lumps presented either in the parasternal region or between the midclavicular and posterior axillary lines. One case presented with an infrascapular lump. Most of the lumps were ill defined and cystic. Chest radiographs in all cases showed a soft tissue swelling without bony involvement. The aspirates were of two types, blood-mixed particles and necrotic material. Four cytologic pictures were seen: (1) epithelioid cell granulomas alone or (2) with necrosis or (3) with both necrosis and acid-fast bacilli (AFB), or (4) necrosis with AFB. AFB were detected in 53.3% of cases. CONCLUSION: If necrotic material is aspirated or if granulomatous inflammation is encountered in soft tissue swellings of the chest wall, particularly in the parasternal and axillary regions, the possibility of intercostal tuberculous lymphadenitis should be considered. PMID- 12585030 TI - A thin-layer, liquid-based pap test for mass screening in an area of China with a high incidence of cervical carcinoma. A cross-sectional, comparative study. AB - OBJECTIVE: To confirm the accuracy of the ThinPrep Pap Test (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) on the basis of histologic diagnosis by biopsy and the detection of human papillomavirus (HPV) DNA in mass screening. STUDY DESIGN: A total of 1,997 women residing in Xiangyuan County, Shanxi Province, P.R.C., an area with a high incidence of cervical carcinoma, were enrolled in this study. We collected exfoliative cervical samples from all subjects into a liquid buffer (Preserv-Cyt [Cytyc]) and utilized for both cytologic screening using the ThinPrep Pap Test and HPV DNA testing. Subsequent colposcopic biopsies were taken on all subjects. All the tests were performed in an independent and blinded fashion. We compared the ThinPrep Pap test with colposcopic biopsy and HPV test. RESULTS: High grade squamous intraepithelial lesions (HSIL) (CIN 2/3) were identified in 74 (3.7%) of 1,993 women adequately assessed, and there were 12 cases of squamous cell carcinoma (SCC). The false negative rate of ThinPrep cytology was 3.2% for biopsy-confirmed CIN 3 and 9.3% for CIN 2. Twenty-seven (87%) of the 31 women with biopsy-confirmed CIN 3 and 12 (100%) of 12 with biopsy confirmed SCC had a diagnosis of either HSIL or greater abnormalities on ThinPrep cytology. In addition, the HPV DNA detection rates offered a good correlation between cytology and biopsy. CONCLUSION: The ThinPrep Pap performed extremely well in this primary screening trial. We found a good correlation between ThinPrep cytology and colposcopic biopsy on detection of HSIL and SCC; cervical specimens collected in ThinPrep liquid buffer serve as a direct test for HPV as well. PMID- 12585032 TI - Thin-layer cytology findings of small cell carcinoma of the lower female genital tract. Review of three cases with molecular analysis. AB - OBJECTIVE: To describe the thin-layer cytology findings of small cell carcinoma of the low female genital tract, with histologic correlation and human papillomavirus (HPV) genotyping. STUDY DESIGN: The authors reviewed the clinical findings, thin-layer cytology and histologic features of small cell carcinoma of the lower female genital tract (cervix or vagina) occurring in three postmenopausal Chinese women at Pamela Youde Nethersole Eastern Hospital, Hong Kong, over a four-year period, from January 1998 to December 2001. Molecular techniques for HPV screening and genotyping using the polymerase chain reaction and restriction fragment length polymorphism were employed on the cytologic specimens. RESULTS: The thin-layer preparations were of moderate to high cellularity. There were loose aggregates of or isolated small round cells with a high nuclear/cytoplasmic ratio, thin but irregular nuclear membrane, hyperchromatic nuclei, inconspicuous nucleoli and scanty cytoplasm. Tumor cell cannibalism was commonly found. Small groups of tumor cells with nuclear molding were noted. There was also obvious tumor diathesis in the background. The necrotic debris was admixed with isolated small round cells, apoptotic bodies and nuclear dust. Associated koilocytosis or squamous intraepithelial lesions were not seen. Histologic examination of the tumor biopsies showed classic features of small cell carcinoma associated with squashing artifacts and vascularized stroma. Molecular analysis revealed the presence of HPV DNA (either type 18 or 16) in all the three liquid-based cytology samples. CONCLUSION: While the cytomorphologic features of small cell carcinoma of the cervix or vagina in thin-layer preparations are slightly different from those in conventional smears, due mainly to the absence of smearing effect, recognition of the subtle but characteristic appearance can enhance the accuracy of the cytologic diagnosis. The association between HPV and primary small cell carcinoma of the lower female genital tract was confirmed by this study. PMID- 12585033 TI - Role of FNAC in metastasizing malignant mixed tumor of the external auditory canal. A case report. AB - BACKGROUND: No previous report of metastasizing mixed tumor (pleomorphic adenoma) of the external auditory canal (EAC) has been described. CASE: A 12-year-old, Chinese girl with a history of mixed tumor of the EAC presented with a locally recurrent, aggressive tumor and metastases to the lung and bone five years later. The primary, locally recurrent and metastatic lung tumor showed epithelial and myoepithelial elements with duct formation, chondromyxoid stroma and mitotic activity in the cellular areas on histology. Fine needle aspiration cytology (FNAC) disclosed the presence of spindle cells blending into chondromyxoid fibrillar ground substance in the recurrent and metastatic lung tumors. CONCLUSION: In primary mixed tumor of the EAC, FNAC plays a useful role in the diagnosis of recurrent and metastatic disease. Its ability to identify ominous features, such as increased mitoses in this case, may be limited by sampling. Since cytology and histology cannot reliably prognosticate, long-term follow up of mixed tumor of the EAC after complete excision is advocated. PMID- 12585034 TI - Fine needle aspiration biopsy of mixed medullary-follicular thyroid carcinoma. A report of two cases. AB - BACKGROUND: Mixed differentiated thyroid carcinomas are rare tumors, difficult to recognize on fine needle aspiration biopsy (FNAB). Most cases are diagnosed only after histologic investigation. CASES: The cases entailed two cytologic samples and a thyroidectomy specimen. Two FNAB thyroidectomy specimens from a 60-year-old man presenting with a solitary thyroid nodule (case 1) were investigated. Both cytologic samples were referred as atypical, with a mixture of features indicating a proliferating follicular lesion but also containing some characteristics of medullary carcinoma. The serum calcitonin level was borderline. Surgery was recommended because of a suspicion of malignancy. The diagnosis of mixed medullary follicular carcinoma was established after a complex histologic investigation. The tumor was encapsulated, with partly microfollicular architecture. Immunohistochemistry was positive for both calcitonin and thyreoglobulin. Electron microscopy from the formol-paraffin block found neurosecretory granules in many cells. The patient was well one year after the operation. One FNAB and thyroidectomy specimen from a 47-year-old woman with long treated lymphoplasmocellular thyroiditis (case 2) was investigated. The tumor in case 2 was diagnosed on FNAB as medullary carcinoma. Only after histologic and immunohistochemical investigation was mixed differentiation proven. CONCLUSION: Mixed differentiated thyroid tumors are a diagnostic challenge on fine needle aspiration. Irrespective of their rarity, they can be suspected if combined features are present. FNAB recognition of the medullary component in both cases was of crucial importance. Nevertheless, definitive diagnosis remains a histologic problem due to the necessity for topographic information. PMID- 12585035 TI - Cytopathologic and clinicopathologic features of ovarian hepatoid carcinoma. A case report. AB - BACKGROUND: Hepatoid carcinoma is a rare ovarian tumor and is thought to be a different histopathologic subtype from hepatoid-type yolk sac tumor based upon its pathologic features. However, the cytopathologic characteristics of ovarian hepatoid carcinoma (OHC) have not been reported previously. We report the clinicopathologic and cytopathologic features and immunoreactivity of a case of OHC. CASE: A 36-year-old woman presented to our department with lower abdominal pain. A left ovarian tumor was found on pelvic examination, magnetic resonance imaging and computed tomography. The tumor was diagnosed as a hepatoid carcinoma of the left ovary based upon the histopathology of the surgically resected specimen. Cytopathologic specimens from a tumor touch preparation of the tumor exhibited pleomorphic tumor cells with abundant cytoplasm. The nuclei contained rough, granular chromatin and large, prominent nucleoli. Several tumor cells were multinucleated. Tumor cells were immunoreactive for alpha-fetoprotein (AFP). Hematoxylin and eosin staining revealed that the tumor cells were in a sinusoidal pattern resembling hepatocellular carcinoma without any glandular formation. The tumor cells were negative for human chorionic gonadotropin while positive for AFP, alpha-1-antitripsin, CA-125 and carcinoembryonic antigen. CONCLUSION: Cytopathologic examination is of considerable aid in the diagnosis of OHC since cytopathologic preparations highlight the characteristic cell pleomorphism. PMID- 12585036 TI - Fine needle aspiration biopsy of neuroendocrine breast carcinoma metastatic to the thyroid. A case report. AB - BACKGROUND: Tumors showing neuroendocrine differentiation arise in a wide range of organs, and metastatic neuroendocrine tumors may be difficult to differentiate from primary tumors. This report describes an unusual case of metastatic breast carcinoma with neuroendocrine differentiation that presented as a solitary thyroid nodule. The diagnosis was made by fine needle aspiration biopsy (FNAB). CASE: A 52-year-old woman presented with a thyroid nodule and bilateral enlarged supraclavicular fossa lymph nodes. FNAB revealed a neuroendocrine carcinoma. Further questioning revealed that the patient had had a breast carcinoma resected eight years previously. The diagnosis of metastatic neuroendocrine breast carcinoma was established by immunocytochemistry. The patient received antiestrogen therapy but subsequently developed skeletal metastases. CONCLUSION: Neuroendocrine carcinomas from various sites show similar cytologic features. In this case, a diagnosis of breast carcinoma metastatic to the thyroid was suggested by the clinical history and confirmed by FNAB with immunocytochemistry. PMID- 12585037 TI - Well-differentiated papillary mesothelioma in the pelvic cavity. A case report. AB - BACKGROUND: Well-differentiated papillary mesothelioma (WDPM) is considered to be a distinct subtype of peritoneal mesothelioma. It occurs in the peritoneum, is most commonly seen in young women and is found incidentally at laparotomy for other indications. Clinically, WDPM is considered to be benign or to have low malignancy potential. CASE: A 48-year-old female with no history of asbestos exposure presented with hypermenorrhea. An operation was performed for adenomyosis, and six papillary nodules, 2 cm or less, were found in the serosa of the pelvic cavity. Peritoneal lavage fluid and imprint material from the tumor were obtained for cytologic examination. The cytologic specimens showed many scattered cells and sheetlike clusters and some papillary clusters. These cells had abundant, polygonal, cyanophilic cytoplasm; clearly outlined borders; and slitlike intercellular spaces. The cell arrangement was orderly. The nuclei were uniform in size, with a single centrally located nucleolus, and there were no binucleated forms or mitosis. There was no increase in chromatin. On the luminal surface of the cells, a brush border was observed. CONCLUSION: It is important to differentiate WDPM from diffuse malignant mesothelioma or other peritoneal malignant tumors to avoid treating them as malignant tumors. PMID- 12585038 TI - Cytology of peritoneal leishmaniasis in a patient with AIDS. PMID- 12585039 TI - Fine needle aspiration cytology of inverted sinonasal papilloma. PMID- 12585040 TI - Blastic extramedullary myeloid cell tumor of the lymph node diagnosed by fine needle aspiration. PMID- 12585041 TI - Cytologic detection of cryptococcosis coexisting with herpes simplex virus infection in sputum: use of liquid-based, thin-layer preparations. PMID- 12585042 TI - Radiation-induced atypical glandular cells mimicking recurrent adenocarcinoma in a posthysterectomy vaginal smear. PMID- 12585043 TI - Giant cells in poorly differentiated (insular) carcinoma of the thyroid. PMID- 12585044 TI - Cytologic features of idiopathic scrotal calcinosis. PMID- 12585045 TI - Multinucleated stromal giant cells in breast lesions. PMID- 12585046 TI - Cardiac tamponade: an unusual presentation of metastatic squamous cell carcinoma of the larynx. PMID- 12585047 TI - [Fracture analysis of a ceramic liner. Is in hip endoprosthesis replacement of ceramic on ceramic components with only one of the corresponding partners justified?]. AB - The good biomechanical and tribological properties, together with the excellent biocompatibility, of ceramic-on-ceramic components, make them a preferential choice for total hip replacement surgery, at least in Europe. We report on a man admitted as an outpatient with painless grating in the hip one year after replacement of a ceramic femoral head, but not of the ceramic inlay. Clinical and radiological findings were indicative of a broken liner. This was confirmed during revision surgery, during which it was replaced by a polyethylene inlay; although the ceramic head appeared intact, it was replaced by a metal head. Inspection of the surface of the broken liner in the scanning electron microscope (SEM) revealed signs of material failure. We recommend careful inspection of ceramic-on-ceramic articulating components during total hip revision surgery and if there is any uncertainty, replacement of both so as to avoid premature failure. PMID- 12585048 TI - [Determination of the tensile strength of superficial passive implant materials]. AB - The crack strength of passivating surface materials or passive layers on electroconductive substrates is determined by the electronic detection of redox reactions at the electrolyte/sample interface. A sudden increase in corrosion current under mechanical tensile loading or bending moments indicates generation or propagation of macro- and micro-cracks in the passivating layer, and exposure of the substrate. A subsequent decrease in the current indicates repassivation. Titanium oxide passivating layers generated by oxygen diffusion hardening (ODH) on titanium show crack formation at a tensile load on the substrate of more than 230 MPa. Repassivating sandwich layers of tantalum and tantalum oxide on steel substrates (AISI 31 6L) generate micro-cracks at more than 300 MPa. The crack formation of the oxide surface materials correlates with the onset of plastic deformation of the substrate. PMID- 12585049 TI - [Surface cleaning of cemented orthopedic implants with conservation of surface properties]. AB - Prior to the analysis of explanted cemented orthopedic implants, complete removal of foreign material, blood and residual tissue and thorough cleaning of the surface without compromising its characteristics is essential. We describe a reprocessing procedure comprising the following 4 steps: 1) retrieval and storage, 2) removal of residual bone cement, 3) chemical cleaning in an enzymatic solution, and 4) mechanical cleaning in an ultrasound bath. PMID- 12585050 TI - [Less is more--a new general practiced oriented form of knee navigation]. AB - Modern navigation technology appears to be acquiring an established place in the fields of orthopedics and orthopedic surgery. This technology helps the surgeon to apply his manual skills with greater precision and thus more effectively, and its positive impact on the quality of surgical treatment has already been demonstrated. The SURGETICS navigation system described in this paper shows that the technology can be adapted to the requirements of daily surgical practice, without compromising its utility for the surgeon and, in the last resort, the patient. Provided that due consideration is given to clinical needs, this technology will surely become a standard tool, not only in European hospitals. PMID- 12585051 TI - [Tensile strength of bone fixation of hydroxyapatite coated Schanz screws of the Heidelberg External Fixation System (HEFS)--comparative torque measurements in clinical use and in cadaver tibia]. AB - It is claimed in the literature that hydroxyapatite(HA)-coated screws of external fixators have superior fixation strength in bone, which is postulated to lead to a substantial decrease in loosening and infection rates. We report on a study of the maximum torque values developed while inserting and removing 30 HA-coated Schanz screws of 8 Heidelberg external fixation systems applied to the tibia to correct leg length differences and axial deformities. The infection rate was determined in accordance with defined criteria, and was found to be about 20% for the HA-coated screws. Screws without infection showed an extraction torque above insertion torque, screws with infection an extraction torque below. A significant correlation (p = 0.05) was seen between infection and decrease in fixation strength (quotient: loosening torque/tightening torque). To exclude the impact of such biological processes as osteointegration and bone remodelling, the clinical results were compared with the torques measured for coated and uncoated Schanz screws in a human cadaveric tibia. A significantly higher fixation strength in bone was found for HA-coated screws in comparison with uncoated screws (p = 0.002). These data warrant a clinical study directly comparing HA-coated and uncoated Schanz screws. PMID- 12585052 TI - [Theory and software of the hexapod external fixator]. AB - Using hexapod kinematics (Stewart platform), an external fixator was developed that can be adjusted in all six spatial degrees of freedom by means of six linear adjusting elements. With such a system, any desired three-dimensional bone movements, for example, fracture reduction or deformity correction, can be effected exactly, without having temporarily to compromise any stability already achieved or to rearrange the construction during treatment. Since the introduction of the hexapod principle by Stewart in 1965, computerized control has been deemed necessary for its application. This paper describes the mathematical basis and the software developed for clinical use. Mathematical procedures are needed for the calculation of the inverse and forward kinematics of the hexapod, and for the description of three-dimensional movements. PMID- 12585053 TI - [Comparison of mechanical properties of orthodontic nickel-titanium wires]. AB - Two packages each, containing 10 wires per package, of different batches of 25 different types of orthodontic archwires made of super-elastic nickel-titanium alloys measuring 0.41 x 0.56 mm2, were investigated. The wires were characterized by obtaining the following measurements at an ambient temperature of 37 degrees: a three-point bending test with the supporting points spaced 10 mm apart, and determination of the torque/bending angle curves using a pure bending test. The force/deflection curves provided the parameters characterizing the super-elastic unloading plateau: average force, slope and endpoint. From the torque/bending angle curves, the parameters average torque, plateau endpoint and the elasticity parameters were determined. Average force (0.8-4.5 N), endpoint (0.2-0.9 mm) and the slope of the unloading plateau (0.2-2.1 N/mm) of the three-point bending test clearly differed for individual wires. Significant differences were also seen for average torque (1.5-11.5 Nmm), unloading plateau endpoint (2.7-20.0 degrees) and elasticity parameters epsilon 4, E4, E5 and E6 in the pure bending test. Individual batches showed only minor differences. The results permit the conclusion to be drawn that super-elasticity is applicable to only a small portion of the wires examined. Although other wires showed super-elastic behaviour, the unloading plateaus has a force level of up to 6 N, and cannot be recommended for orthodontic application. The super-elastic plateau is often of use only for deflections greater than 1.5 mm. The use of super-elastic archwires made of nickel-titanium alloys makes sense only when the elastic properties of the respective wires are known. This makes the provision by the manufacturer of relevant data on the elastic properties of wires a necessity. PMID- 12585054 TI - Anthrax and the voice of reason. PMID- 12585055 TI - Why are nurses virtually absent? PMID- 12585057 TI - The decline of homeopathy. PMID- 12585058 TI - Hyperbaric oxygen therapy. PMID- 12585059 TI - Hyperbaric oxygen therapy. PMID- 12585060 TI - Hyperbaric oxygen therapy. PMID- 12585062 TI - The naked decimal point. And eight other common errors that can be avoided. PMID- 12585063 TI - Heart failure: pathophysiologic mechanisms. PMID- 12585064 TI - Managing patients with heart failure. PMID- 12585065 TI - Home management of the failing heart. Inotropic therapy in the outpatient setting. PMID- 12585066 TI - Emergency. Anthrax. PMID- 12585067 TI - Health care reform: what went wrong? PMID- 12585068 TI - The patient's report of pain. PMID- 12585069 TI - A sound partnership for end-of-life care. PMID- 12585070 TI - Nursing malpractice: protect yourself. PMID- 12585071 TI - Carbon monoxide poisoning in the pediatric population. PMID- 12585072 TI - Hartford LIFE STAR. Care in the New England sky. PMID- 12585073 TI - Statewide air medical transports for Massachusetts. AB - In 1997, the Massachusetts Department of Public Health (MDPH) established a process to centralize air medical transport information. This database is one of the first statewide, population-based sources for civilian rotary-wing air medical transports (U.S. Coast Guard, police, and military missions are not included). The purpose of this database is to facilitate MDPH review of air medical transport service utilization, with input from a multidisciplinary committee. This article discusses the challenges in producing uniform data from multiple service submissions and presents aggregate "baseline" utilization information for 1996. These data served as a starting point for later studies using data linkage. This indexed article is the first to report statewide, population-based data for all types of air medical helicopter transports. The only other indexed "statewide air medical transport" paper focused on scene transports to trauma centers in Pennsylvania. A previous article by the authors in the July-September 2000 Air Medical Journal provided an overview of air medical transports for fatal motor vehicle crashes for 1 region of the state. PMID- 12585074 TI - Flight team management of in-place endotracheal tubes. AB - INTRODUCTION: Unintended misplacement or removal of the endotracheal tube (ETT) complicates the care of up to 18% of intubated patients. This project analyzed the incidence of such complications in patients transported by a flight program. METHODS: 9-month analysis of all intubated patients transported by the flight team. RESULTS: 340/926 patients transported were intubated. One extubation was unplanned and no patients were delivered to the receiving hospital with an esophageal or endobronchial ETT placement. After initial examination, 19/241 ETTs placed before flight team arrival were repositioned. Rates of misplacement on arrival at the receiving hospital and of unplanned extubation were significantly lower than those reported in the EMS or critical care literature. CONCLUSION: Flight teams have very low rates of unplanned extubation or undetected ETT misplacement when transporting intubated patients. PMID- 12585075 TI - [Depressive effect of tetrandrine on collagen synthesis in vascular smooth muscle cells]. AB - AIM: To clarify whether collagen deposition and collagen synthesis are increased in renovascular hypertensive rats (RHR) and whether these are affected by treatment with tetrandrine (Tet). METHODS: Collagen deposition was detected by electron microscope and evaluated by the amount of hydroxyproline in aorta of sham-operated rats (Sham), renovascular hypertensive rats (RHR), and Tet treated RHR. The aortic vascular smooth muscle cells (AVSMCs) were isolated and cultured. The collagen synthesis in cultured AVSMCs was detected by [3H]-proline incorporation. RESULTS: The ultrastructure of aorta suggests that AVSMCs in RHR had been transferred from contractile phenotype to synthetic phenotype. Compared with sham-operated rats, the hydroxyproline content in aorta was increased by 36.6% in RHR, while it was reduced by 18.7% in Tet treated RHR (P < 0.01, vs RHR). Tet (0.1-10 mumol.L-1) treatment in vitro induced a concentration dependent depression [3H]-proline incorperation stimulated by NE or Ang II in AVSMCs from either Sham or RHR. CONCLUSION: The depressive effects of tetrandrine on collagen synthesis and deposition in arteries may contribute to the regression of vascular remodeling. PMID- 12585076 TI - [The expression levels of GIRK1 and IRK1 in memory deficit rats]. AB - AIM: To study the expression level of GIRK1 and IRK1 in cerebral cortex after aggregated beta-amyloid peptide25-35 injection into the rat cerebral ventricle for different times. METHODS: To test the spatial learning and memory in Morris water maze. To observe GIRK1 and IRK1 expression levels by RT-PCR. RESULTS: In Morris water maze task, the latencies of beta-amyloid peptide treated rats were longer than those of the sham rars in the 4, 5, 8, 10 training periods ("student" t-test, P < 0.05). The expression levels of GIRK1 in beta-amyloid peptide injection day 1 and day 3 groups are decreased significantly compared with sham group (Tukey's test, P < 0.05) and there is no difference between beta-amyloid peptide injection day 17 group and sham group. IRK1 expression levels are no difference among the four groups. CONCLUSION: Injection of aggregated beta amyloid peptide25-35 into rat cerebral ventricles can induce memory and potassium channel expression changes. Beta-amyloid peptide induced inhibition of GIRK1 expression may have some relations with brain dysfunction. PMID- 12585077 TI - [Bepridil inhibition on the delayed rectifier K+ currents in thyroxine induced hypertrophied guinea pig ventricular myocytes]. AB - AIM: To study the effects of bepridil on the rapidly activating component (IKr), the slowly activating component (IKs) of the delayed rectifier potassium current and the inward rectifier potassium current (IK1) in hypertrophied guinea pig ventricular myocytes. METHODS: The whole cell patch clamp techniques were used. RESULTS: In hypertrophied guinea pig ventricular myocytes, bepridil 30 mumol.L-1 markedly inhibited IKr and IKs (by 20.9% and 27.2% at 0 mV and mV, respectively). The effect of bepridil on IKs was larger than on IKr. Bepridil 30 mumol.L-1 also significantly inhibited the inward component of IK1 (by 15.1% at +100 mV), but the reverse potential of IK1 was unaffected. Bepridil (1-100 mumol.L-1) was shown to inhibit IKr and IKs in a concentration-dependent manner. Their IC50 were 46.7 mumol.L-1 and 23.8 mumol.L-1, respectively. CONCLUSION: Bepridil inhibit IKr, IKs and IK1 in hypertrophied guinea pig ventricular myocytes, which may be important in understanding the antiarrhythmic effects of this drug. PMID- 12585078 TI - [Effect of dimethyl diphenyl bicarboxylate (DDB) on 9-amino-1,2,3,4 tetrahydroacridine-induced hepatotoxicity in mice]. AB - AIM: To establish an acute model of THA-induced hepatotoxicity in mice, and observe the effects of DDB on THA-induced liver damage. METHODS: After a single oral dose of THA (56 mg.kg-1), body temperatures, liver MDA content and serum ALT were measured within 12 h. The activities of mice brain acetylcholinesterase, liver microsomal 7-ethoxyresorufin deethylase, uridine-5'-diphosphoglucuronic transferase, and mitochondria potential change were also observed. RESULTS: The alteration of mice body temperature, and the elevation of serum ALT, liver MDA content, and mitochondria potential induced by THA were significantly inhibited by DDB pretreatment. The microsomal 7-ethoxyresorufin deethylase activity was induced by DDB too. On the other hand, The inhibiting effects of THA on mice hippocampus and cortex acetylcholinesterase in vitro and in vivo were not influenced by DDB treatment. CONCLUSION: The toxic effect of THA on mice liver was significantly reduced by DDB. These results demonstrate that the protective action of DDB may attribute to its regulation on enzymes involved in THA metabolism, and its protective effect against mitochondria injury caused by THA. Therefore, DDB may be a potential liverprotector against THA-induced hepatotoxicity during dementia therapy. PMID- 12585079 TI - [Mechanism of apoptosis induced by squamocin in leukemia cells]. AB - AIM: To investigate the mechanism of apoptosis of HL60 cells induced by the annonaceous acetogenin, squamocin. METHODS: Induction of apoptosis was determined through Hoechst33258 dye staining and DNA agarose gel electrophoresis. Expression of the proteins was detected using Western blot analysis. Caspase-3 activity was detected using caspase-3 kit. RESULTS: Treatment of HL-60 cells with squamocin resulted in extensive nuclear condensation, DNA fragmentation, cleavage of the death substrate poly(ADP-ribose) polymerase (PARP) and induction of caspase-3 activity. Pretreatment of HL-60 cells with caspase-3 specific inhibitor DEVD-CHO prevented squamocin-induced DNA fragmentation, PARP cleavage and cell death. Stress-activated protein kinase (SAPK/JNK) was activated after treatment with squamocin in HL-60 cells. CONCLUSION: These results suggest that apoptosis of HL 60 cells induced by squamocin require caspase-3 activation, and could be related to SAPK activation. PMID- 12585080 TI - [Synthesis and vasodilative activities of alpha-phenylcinnamide]. AB - AIM: To search for compounds having strong vasodilating effect. METHODS AND RESULTS: The mixed anhydride and alpha-phenylcinnamyl halide were used. Twenty two alpha-phenylcinnamide derivatives were synthesized. The strutures were elucidated on the basis of MS and 1HNMR. Compounds 1-22 are new compounds. CONCLUSION: Vasodilative activity assays were conducted for the target compounds and the results indicated that several compounds (3, 9 and 11) demonstrated superior pharmacological profiles to the lead compound, among which compound 11 was further evaluated and found to be indicative of potential KCO activity. Preliminary SAR of alpha-phenylcinnamides was discussed herein. PMID- 12585081 TI - [Three dimensional quantitative structure activity relationship of P450(17) alpha inhibitors of 17-substituted steroids]. AB - AIM: To develop a three dimensional quantitative structure activity relationship (3D-QSAR) model and gain further insights into the requirements for potential P450(17) alpha inhibitors. METHODS AND RESULTS: A predictive 3D pharmacophore model was established based on comparative molecular field analysis (CoMFA). The correlation between the activities and structures was significant with cross validated value (R2cv), non-cross-validated value (R2) and standard error of estimate (SEE) of 0.538, 0.799 and 0.257, respectively. According to this model, the predicted inhibition activities of three compounds synthesized in our laboratory were compatible to actual activities. CONCLUSION: This model would contribute to the understanding of the interaction between the inhibitors and P450(17) alpha and rational design of novel lead molecules. PMID- 12585082 TI - [Studies on the chemical constituents of Zanthoxylum dimorphophyllum Hemsl. var spinifolium Rhed. et Wils]. AB - AIM: To study the chemical components from the dried roots of Zanthoxylum dimorphophyllum Hemsl. var spinifolium Rhed. et Wils. METHODS: Modern chromatography was used to isolate chemical components and the compounds isolated from Zanthoxylum dimorphophyllum Hems1. var spinifolium Rhed. et Wils were identified by various spectral methods. RESULTS: Five compounds were isolated and identified as 6-(2',3'-dihydroxy-3'-methybutyl)-7-acetoxy-2H-1-benzopyran-2-one (I), 6-(2',3'-dihydroxy-3'-methybutyl)-7-(beta-D-glucopyranosyloxy)- 2H-1 benzopyran-2-one (II), hexacosanoci acid (III), aurepten (IV), 6-[6'-O-(-deoxy alpha-L-mannopyranosyl)-beta-D-benzopyran-4-one (V). CONCLUSION: All of the above compounds were isolated from the plant for the first time, among them, I is a new compound. PMID- 12585083 TI - [An abzyme to catalytize the deiodination of thyroxine]. AB - AIM: To mimic an important family of selenoenzymes in organism-thyroxine (T4) deiodinases and prepare a selenium-containing abzyme catalyzing deiodination of T4. METHODS: A anti-T4 monoclonal antibody was generated by hybridoma methodology and converted into a selenium-containing abzyme by the method of chemical modification. The catalytic activity of the enzyme was measured by RIA method. RESULTS: The abzyme displayed a marked activity of catalyzing deiodination of T4 and a higher specificity to the substrate T4 than that of natural enzyme, and the double reciprocal plots of the initial rates of T3 formation vs. T4 concentration yielded a family of parallel lines. The catalytic activity could be sensitively inhibited by 6-propyl-2-thiouracil (PTU), a competitive inhibitor for dithiothreitol (DTT). CONCLUSION: An abzyme with the diodination activity was first prepared and the reaction mechanism of the enzyme was bisubstrate ping-pong one. PMID- 12585084 TI - [Glucosides from Pleurospermum rivulorum]. AB - AIM: To study the chemical constituents of Pleurospermum rivulorum. METHODS: To separate the constituents of P. rivulorum by using various kinds of chromatography and identify their structures on the basis of spectral analysis. RESULTS: Four glucosides were isolated from P. rivulorum. Their structures were established as marmesinin (I), tert-O-beta-D-glucopyranosyl-heraclenol (II), 1'-O beta-D-glucopyranosyl (2R, 3S)-3-hydroxynodakenetin (III) and 8-hydroxy-5-O-beta D-glucopyranosyl-psoralen (IV) based on the analysis of spectral data of IR, UV, MS, 1D and 2D-NMR. CONCLUSION: Compounds III and IV are new compounds. All of the four compounds were isolated from this plant for the first time. PMID- 12585085 TI - [Characterization and recognition key components in Astragalus membranaceus]. AB - AIM: To separate, identify and quantify multi-components in complex traditional Chinese medicine (TCM) Astragalus membranaceus by unified analysis for systematic study of its relative contents. METHODS: The analytical method was reverse-phase HPLC/photodiode array detector. Based on the comparison of UV spectra, software of UV database to assist recognition has been established. Key components have been found and recognized in two reference Astragalus membranaceus and three samples from different sources. With the unified analytical project, relative peak heights of key components were recognized and contrasted. RESULTS: The water soluble components can be separated by solvent system A, a gradient program with mobile phase of methanol-water. The liposoluble components can be separated with the solvent system B, a gradient program of mobile phase of 2-propanol acetonitrile-water. UV detection wavelength was set at 202 nm and the flow rate was 1 mL.min-1 on a Hypersil ODS column. Twenty-nine components in the four Astragalus membranaceus were recognized using characteristic parameters of chromatography and UV spectroscopy. All of the 29 key components in two reference samples and three samples from different sources were shown in their chromatograms to be the same. The concentrations of some the key components in different samples were similar while those of the others were different. The chromatographic peaks of the reference Astragalus membranaceus and two samples were matched with each other. Many different fingerprints were found and differences were shown on their chromatograms. Not only there were differences among three samples from different source, but also there were differences in the two reference samples. CONCLUSION: To find out the key components and recognize them are the basis of quantitative recognition and are very useful when standards are not available. The method of multi-component quantitative recognition in Astragalus membranaceus was developed. The method is convenient and reproducible. It can be widely used for the quality control of the Astragalus membranaceus. PMID- 12585087 TI - A new crystal form of nateglinide. AB - AIM: To find a new crystal structure of S-form in nateglinide [N-(trans-4 isopropylcyclohexylcarbonyl)-D-phenylalanine]. The XRD, IR patterns and data were given and the melting point was determined. METHODS: Phase analysis was performed by X-ray powder diffraction, IR and elemental analysis and the melting point was determined by differential scan calorimetry. RESULTS: S-form nateglinide is different from H-form or B-form. The melting point is 172.04 degrees C. CONCLUSION: The S-form nateglinide is a new crystal form. PMID- 12585086 TI - [In vitro metabolic studies of the novel anti-anxietic drug AF-5 and its metabolites in human liver microsome incubation system]. AB - AIM: To study the metabolism of a novel anti-anxietic drug AF-5 and its metabolites (I, II) in human liver microsome incubation system. METHODS: Human liver microsomes were prepared, the enzyme activity was determined to be 8.79 mg.mL-1 by Lowry's method. The human liver microsome incubation system consisted of: human liver microsomes 2 mg.mL-1, glucose-6-phosphate (G-6-P) 0.01 mmol.mL-1, glucose-6-phosphate dehydrogenase (G-6-PDH) 1 U.mL-1, magnesium chloride (MgCl2) 4.0 mumol.mL-1, coenzyme II in oxidized form (NADP) 0.5 mumol.mL-1, and coenzyme I in reduced form (NADH) 1.0 mumol.mL-1. Two milligrams of AF-5 solubilized by Tween 80 was then added, the mixture was diluted to 5 mL with Tris-HCl solution and the mixture was incubated in a 37 degrees C water bath with shaking. Oxygen was passed over the liquid surface for 0.5 min every 20 minutes. The incubation was carried out for 40 min and 100 min respectively. Three volumes of ethyl ether were added to stop the metabolism, and more ethyl ether was used to extract the metabolites for 3 times. The ether extracts were pooled together, dried with anhydrous sodium sulfate, then evaporated to dryness. The residue was dissolved in 0.5 mL n-hexane and analyzed by GC/MS under the following conditions: 150 degrees C (1 min)[formula: see text]180 degrees C (1 min)[formula: see text]260 degrees C (2 min), in the total ion current mode, EI: 70 eV, interface temperature: 250 degrees C, ion source temperature: 200 degrees C. RESULTS: Two major metabolites were found and identified in this incubation system, and demonstrated that the in vitro metabolic pathway was that the carbon 4 was first oxidized to hydroxyl group, then further oxidized to a carbonyl group. CONCLUSION: In human liver microsome incubation system AF-5 was completely metabolized in 100 min to the hydroxy derivative I and carbonyl derivative II, with hydroxymetabolite as the major metabolite. Metabolite I was further transformed to metabolite II, which was not metabolized any further by the human liver microsomes. PMID- 12585088 TI - [Pharmaceutical evaluation of fast-disintegrant tablet containing nicorandil loaded particles]. AB - AIM: To improve the bioavailability and taste of fast-disintegrating tablet (FD tablet) containing nicorandil-loaded particles. METHODS: A FD tablet containing nicorandil-loaded particles with 1%-4% croscarmellose sodium in addition of D mannitol and lactose (9:1) was prepared and the dissolution and absorption characteristics were examined, in comparison with FD tablet and commercial tablets of nicorandil. In vivo absorption of nicorandil from FD tablet was evaluated in beagle dogs. RESULTS: The disintegration time of FD tablets containing 1% croscarmellose sodium with 6 mm and 10 mm in diameter were about 12 and 23 seconds, respectively. When nicorandil-loaded particles consist of myristyl alcohol and stearyl alcohol were put in FD tablet, nicorandil release from FD tablet continued until 6 h while nicorandil release from Sigmart and FD tablet containing nicorandil crystals finished within 5 min. In vivo absorption of nicorandil from Sigmart and FD tablet containing nicorandil crystals was very similar after oral administration in beagle dogs and no statistic difference in AUC, Tmax, Cmax was observed between these tablets. However pharmacokinetics parameters of nicorandil after oral administration of FD tablet containing nicorandil-loaded particles showed that nicorandil was delivered into the body at a suitable absorption rate with similar AUC, delayed Tmax and lower Cmax. CONCLUSION: The reports suggest that the modification of properties of myristyl alcohol and stearyl alcohol released from the drug-loaded particles system would lead to more acceptable bioavailability of the system. However, The formulation of particles and may have a masking effect against the bitter taste and irritation of the drug. PMID- 12585089 TI - [Study on third-type immunoliposomes loaded drugs and the targeting in vitro and in vivo]. AB - AIM: To study the preparation, targeting and pharmacodynamics of third-type immunoliposome loaded anticancer drugs. METHODS: The monoclonal antibody of human bladder cancer was combined with the terminal of PEG-COOH (polyethyleneglycol carboxylic acid) that make the liposomes not only prolong circulation by the membrane protection of PEG, but also target by spreading the antibody on the liposomes surface. That was the third type immunoliposomes. According to this scheme, the IML-ADM (immunoliposome carried adriamycin) wes prepared in which ADM entrapment was efficient and stability was high and the antibody activity was kept. RESULTS: The % survival of the targeting EJ cells treated with IML-ADM (ADM = 45.45 micrograms.mL-1) was 4.3% +/- 1.0%, but 72% +/- 6% for non-targeting LOVO cells in vitro; the tumor weight in nude mice which were implanted by EJ cells after 27 days were (39 +/- 25) mg, (135 +/- 32) mg, (598 +/- 240) mg treated by IML-ADM, SSL-ADM (steric stable lipsomes carried Adriamycin) and normal saline, respectively, in vivo. CONCLUSION: The results confirmed that the immunoliposme mediated targeting anticancer drug is a feasible way. PMID- 12585090 TI - [Investigation of the bioadhesion of carbopol and hydroxypropyl methylcellulose to rat gastrointestinal mucosa in vivo and in vitro]. AB - AIM: To investigate bioadhesion of carbopol and hydroxypropyl methylcellulose of different viscosity grade to rat gastrointestinal mucosa. METHODS: The maximum adhesion force has been adopted as an index of adhesive evaluation in vitro, gastro-emptying rate and migrating length in intestine have been adopted as an index of adhesive evaluation in vivo. RESULTS: In vitro adhesion studies indicated that the adhesive force of low-vicosity materials was stronger than that of high-vicosity materials, while the transferation rate of high-vicosity materials is significantly higher compared to that of low-vicosity materials in vivo. CONCLUSION: Adhesive force of materials gives no correlation with rate of the materials in rat; Cb934 have the optium bioadhesion among the studied materials and can be used as the preferred adjuvant in oral bioadhesion preparation. PMID- 12585091 TI - [Studies on the anthraquinones of Cassia siamea]. AB - AIM: To study the anthraquinone constituents of the stem of Cassia siamea. METHODS: The compounds were isolated by chromatography on silica gel, MHPLC, and identified on the basis of spectral analysis including IR, EI-MS, FAB-MS, 1HNMR, 13CNMR and DEPT. RESULTS: Three compounds were isolated and identified as: chrysophanol (I), chrysophanol-1-O-beta-D-glucopyranoside (II) and 1-[(beta-D glucopyranosyl-(1-->6)-0-beta-D-glucopyranosyl) oxy]-8-hydroxyl-3-methy-9,10 anthraquinone (III). CONCLUSION: III is a new compound, II was obtained from this plant for the first time. PMID- 12585092 TI - [Interaction between protein and ciprofloxacin]. AB - AIM: To study the interaction between ciprofloxacin and BSA in physiological condition by fluorescence spectroscopy. METHODS: The affection of drug to the protein conformation was investigated. The binding constant between drug and BSA from a double reciprocal Lineweaver-Burk plot was determined and the main sort of binding force was found according to the thermodynamic parameters. RESULTS: The binding constants between BSA and ciprofloxacin at 26 degrees C and 45 degrees C are about 10(4). At 26 degrees C, the thermodynamic parameters of reaction between BSA and ciprofloxacin are delta H = -49.13 kJ.mol-1, delta G = -26.45 kJ.mol-1, delta S = -75 kJ.mol-1. The maximum wavelength of the synchronous fluorescence spectra of BSA moved from 279 nm to 289 nm with the increasing of the amount of ciprofloxacin. CONCLUSION: There exists fluorescence energy transfer between BSA and ciprofloxacin. The main sort of binding force between BSA and ciprofloxacin is Van der Waals' interaction. Ciprofloxacin can be deposited and be transported by serum protein in vivo. Ciprofloxacin affects the protein conformation. PMID- 12585093 TI - [A new antitumor agent--endostatin]. PMID- 12585094 TI - [Application and perspective of microdialysis sampling in metabolism study of drugs]. PMID- 12585095 TI - [The days followed veteran traditional doctors for clinical study]. PMID- 12585096 TI - [Clinical study on effect of Agkistrodon antithrombogenase in auxiliary treatment of rheumatoid arthritis]. AB - OBJECTIVE: To explore the clinical effect of Agkistrodon antithrombogenase (AAT) in the treatment of rheumatoid arthritis (RA) and its possible mechanism. METHODS: Besides the conventional non-steroid anti-inflammatory agents and disease-modifying anti-rheumatic drug, patients were treated supplementally with intravenous injection of AAT. The intracutaneous test showed allergic to AAT patients were treated with Salvia injection and taken as control group. Changes of related clinical indexes in the two groups were observed. RESULTS: After 3 weeks treatment, condition of patients in both groups were improved clinically in joint swollen index, joint tenderness index, morning stiffness time, pain assessment (VAS) and health assessment questionnaire (HAQ) on daily life activity as well as ESR level (P < 0.05 or P < 0.01), with the VAS, HAQ and fibrinogen levels more significantly improved than those of control (P < 0.05 or P < 0.01), and the total effective rate higher in the AAT treated group than those in the control group (P < 0.05). CONCLUSION: AAT has good effect on easing clinical symptoms of RA possibly through anti-inflammation and improving the microcirculation with less toxic and adverse reaction, so is worthy of recommendation. PMID- 12585097 TI - [Study on effect of Cordyceps sinensis and artemisinin in preventing recurrence of lupus nephritis]. AB - OBJECTIVE: To observe the effect of Cordyceps sinensis and artemisinin in preventing recurrence of lupus nephritis (LN). METHODS: Sixty-one LN patients, who had no activities by corticosterone and cyclophosphamide (CTX) impacting therapy were randomly divided into two groups. The 31 cases in the treated group were given Cordyceps powder 2-4 g/d before meal and artemisinin 0.6 g/d after meal in three portions orally taken for 3 years. The 30 patients in the control group were treated with tripterygiitotorum and/or Baoshenkang tablet. The consecutive observation lasted for 5 years to monitor the clinical manifestations of lupus and laboratory indexes including blood creatinine, creatinine clearance rate (CCr) and antinuclear antibodies (ANA). RESULTS: The therapeutic effect showed markedly effective in 26 cases (83.9%), effective in 4 (12.9%) and ineffective in 1 (3.2%) in the treated group, while in the control group, the corresponding numbers were 15 (50.0%), 8 (26.7%) and 7 (23.3%), the difference between the two groups in markedly effective rate was significant (P < 0.01). In the treated group, C3 level was stabilized at above 1.21 +/- 0.20 g/L, which was over the normal range, CCr was unchanged as compared before and after treatment, which was significantly different from that in the control group. Moreover, the side-effects occurred in the treated group was less. CONCLUSION: Cordyceps and artemisinin could prevent the recurrence of LN and protect kidney function. PMID- 12585098 TI - [Clinical observation on effect of yiqi yangyin principle on corticosteroid withdrawal in patients with systemic lupus erythematosus at remission stage]. AB - OBJECTIVE: To observe the therapeutic effect of Yiqi Yangyin principle (YQYY, the treating principle in TCM to supplement Qi and nourish Yin) on corticosteroid withdrawal in patients with systemic lupus erythematosus (SLE) in remission stage and its influence on some immune parameters. METHODS: The SLE patients were divided into two groups, 30 in the treated group and 10 in the control group, who were treated by conventional method with corticosteroids and/or immunosuppressant in acute progressive stage, and YQYY was added to the treated group in remission stage. RESULTS: The total effective rate was 93.3% in the treated group and 90.0% in the control group, comparison between the two groups showed significant difference by Ridit test (P < 0.05). The immune parameters, IgG and C3 were significantly improved after treatment in the treated group (P < 0.01), but changed insignificantly in the control group. The maintaining dose of prednisone used in the two groups was 7.08 +/- 5.26 mg/d and 11.72 +/- 6.48 mg/d respectively, the amount used in the treated group was significantly lower than that in the control. CONCLUSION: Using mainly YQYY Principle to treat SLE in remission stage could withdraw the corticosteroid smoothly, relieve symptoms and improve immune function. PMID- 12585099 TI - [Clinical effect of Tripterygiitotorum combined with prednisone and its effect on serum IL-6 level in treating patients with myasthenia gravis]. AB - OBJECTIVE: To explore the clinical effect of Tripterygiitotorum (T II) combined with prednisone in treating patients with myasthenia gravis (MG) and the changes of immune function after treatment. METHODS: Sixty-eight patients with MG were randomly divided into two groups, the 36 patients in the treated group were treated with T II plus prednisone and the 32 in the control group were treated with prednisone alone. The therapeutic effect, serum interleukin-6 (IL-6) and peripheral B lymphocyte levels were observed. RESULTS: The therapeutic effect in the treated group was significantly higher than that in the control group (P < 0.05). There were significant decrease in serum IL-6 and peripheral B lymphocyte in both groups after treatment (P < 0.05), with the decrements more significant in the treated group (P < 0.05). CONCLUSION: The therapy of T II plus prednisone is superior to that of prednisone alone in treating MG, it has a more potent effect of immunosuppression. PMID- 12585100 TI - [Clinical and experimental study on retardation of immunosenescence by kidney tonifying principle]. AB - OBJECTIVE: To observe the clinical curative effect of Kidney tonifying method on retardation of immunosenescence and corresponding experimental study. METHODS: A randomized double blind placebo-controlled trial was used (RCT) on 22 pairs of aged subjects to elucidate the effect of Kidney tonifying recipe on the peripheral T-lymphocyte apoptosis and the Fas/FasL gene expression in them. In rats experimental study, the effects of two kinds of Chinese recipes (Kidney tonifying recipe and blood circulation promoting recipe) on the same parameters as in clinical study as well as on cell apoptosis and gene expression regulation in old rats were also observed. RESULTS: Clinical study showed that after treatment, the percentage of T-lymphocyte apoptosis and the FasL gene expression in the Kidney tonifying group of aged subjects were lower than those in the placebo group (P < 0.01). Animal experiment showed the same result as shown in clinical study in Kidney tonifying recipe treated rats, but not shown in those treated with blood circulation promoting recipe statistically. CONCLUSION: Kidney tonifying principle has down-regulating effect on the transcription of apoptotic gene in both aged persons and old rats, this is one of the molecular mechanisms of Kidney tonifying method in decreasing over-apoptosis in aged subjects and old rats. PMID- 12585101 TI - [Clinical observation on treatment of postpartum hemorrhage by xuesaitong soft capsule]. AB - OBJECTIVE: To observe the effect of Xuesaitong (XST) soft capsule in treating postpartum hemorrhage. METHODS: Sixty pregnant women were randomly divided into the treated group (A) and the control group (B), 30 cases in each, they were treated with XST and placebo respectively for 7 days to observe the changes of hemorrhagic conditions. RESULTS: After delivery, the hemoglobin, RBC count, hematocrit and cardiac index were lower at 3rd day than those before delivery in both groups (P < 0.05 or P < 0.01), reticulocyte count lowered insignificantly as compared with that before delivery in group A but significantly in control group B; the bleeding time shortened significantly in group A (P < 0.05) but unchanged in group B; significant change of blood viscosity only found in group B (P < 0.05). The amount of bleeding 30 min, 1 hr, 2 hrs and 24 hrs after delivery in group A was all lower than that in group B respectively (P < 0.01 or P < 0.05). Involution of uterus in group A at 2nd, 4th and 7th day after delivery was all better than that in group B respectively (P < 0.05). CONCLUSION: XST has definite effect in decreasing postpartum hemorrhage, it also shows effects in promoting involution of uterus, reducing lochia and accelerating recovery of parturients. PMID- 12585102 TI - [Study on stroke syndrome of Chinese medicine and its relationship with blood hypercoagulative status and insulin resistance in diabetic ischemic stroke patients]. AB - OBJECTIVE: To study the relationship of TCM Syndromes, involving Viscera Syndrome and involving Meridian Syndrome, with blood hypercoagulative state and insulin resistance in patients with diabetic ischemic stroke. METHODS: Insulin sensitivity index (ISI) could reflect the insulin resistance, and those reflecting blood hypercoagulative state parameters such as platelet agglutination test (PAgT), fibrinogen (FG), in vitro thrombus length (VTL) and activating partial thrombinogen time (APTT), were used to expound and prove the relationship with the stroke syndrome. RESULTS: The symptom scores, PAgT, VTL, FG levels in patients involved with Viscera Syndromes (including those of Wind Syndrome, Phlegm-Dampness Syndrome, Heat-Fire Syndrome and Yin-deficiency with Yang-excess Syndrome) were significantly higher than those in patients involved with Meridian Syndromes (P < 0.05 or P < 0.01), while the ISI level in the former was lower than that in the latter (P < 0.01). CONCLUSION: There is close relationship between blood hypercoagulative state, insulin resistance and TCM Stroke Syndromes in patients with diabetic ischemic stroke. PMID- 12585103 TI - [Clinical observation of ruangan suopi tablet in treating chronic hepatitis B caused liver cirrhosis]. PMID- 12585104 TI - [The intervention effects of Angelica sinensis, Salvia miltiorrhiza and ligustrazine on peritoneal macrophages during peritoneal dialysis]. AB - OBJECTIVE: To study the effect of Angelica sinensis, Salvia miltiorrhiza and Ligustrazine on function of peritoneal macrophages during peritoneal dialysis. METHODS: Peritoneal macrophages of mice were cultured in culture medium (control), peritoneal dialysate (PD), drugs contained PD containing Angelica, Salvia and Ligustrazine combined (PD-ASL) or separated (PD-A, PD-S, PD-L) with concentration of 2 micrograms/ml, 10 micrograms/ml and 100 micrograms/ml, separately for 24 hrs. The nitric oxide (NO) content, methyl thiazolyl tetrazolium (MTT) reducing capacity (MTT-RC) and phagocytosis capacity of macrophages were determined and compared. RESULTS: NO content and MTT-RC of macrophages cultured in PD group were significantly lower than those of the control (P < 0.01), as compared with those in drug contained PD groups, the NO content in the PD-L group and the MTT-RC in the PD-ASL group were higher significantly (P < 0.01). The phagocytosis capacity and NO content in the PD-ASL group were raised along with the increased concentration of drug in PD. CONCLUSION: Administering Chinese herbal medicine during peritoneal dialysis has important significance in improving the defense function of peritoneal macrophages, reducing the incidence of peritonitis and enhancing the therapeutic effect of peritoneal dialysis. PMID- 12585105 TI - [Effect of buyang huanwu decoction and its active regions combination on brain heat shock protein 70 expression in gerbils after cerebral ischemia/reperfusion]. AB - OBJECTIVE: To compare the effects of Buyang Huanwu Decoction (BYHWD) and its active regions combination on brain heat shock protein 70 (HSP70) expression. METHODS: Cerebral ischemia model gerbils produced by ligating bilateral common carotid artery were injected intraperitoneally by BYHWD and its active regions combination separately. HSP70 mRNA expression was determined by Northern blot test and protein expression determined by Western blot test 48 hrs after ischemia for 15 min and reperfusion. RESULTS: The level of HSP70 mRNA and protein expression in ischemic/reperfusion brain tissue increased significantly. BYHWD and its active regions combination could decrease the HSP70 mRNA expression obviously, the inhibition effect of BYHWD was more significant than that of its active regions combination, but neither of them showed any effect on HSP70 protein expression. CONCLUSION: BYHWD and its active regions combination might down-regulate HSP70 mRNA expression in ischemic brain tissue through its anti ischemic effect. PMID- 12585106 TI - [Experimental study on effect of xiaolong tongbi on nitric oxide synthase and epithelial cell apoptosis in prostate of rats]. AB - OBJECTIVE: To explore the effect of Xiaolong Tongbi (XLTB), a Chinese herbal preparation, on nitric oxide synthase (NOS) containing nerve and epithelial cell apoptosis. METHODS: NOS containing nerve level of various groups prostate was determined by NADPH histochemical staining and morphological quantitative analysis, and the cell apoptosis rate in rats prostatic epithelium was tested immunohistochemically with TUNEL method. RESULTS: After 3 weeks treatment with high dose XLTB, the length density of NOS contained nerves in prostate tissue of rats was 0.113 +/- 0.023, which was significantly different to that in the other 3 groups (low dose XLTB, Finasteride and testosterone) respectively (P < 0.01). Cell apoptosis reached its peak at the day 7 of experiment in XLTB treated groups (9.27%), and Finasteride treated group (5.65%), which lowered slightly at day 14 and got some recovery at day 21. Castration induced cell apoptosis began at day 7 and reached its peak at day 14, which was significantly different from that in the other groups. CONCLUSION: XLTB could increase the NOS contained nerve level and accelerate the apoptosis of epithelial cell in prostate tissue of simulating benign prostatic hyperplasia rats. PMID- 12585107 TI - [Effect of shengji huayu recipe and its disassembled formulae on type I and III collagen synthesis in wound healing fibroblasts]. AB - OBJECTIVE: To explore the mechanism of Shengji Huayu Recipe (SJHYR) and its 3 disassembled formulae in accelerating wound healing toward skin restoration. METHODS: Using in vitro cultured fibroblasts from granulation tissue of wound and compared with the normal skin fibroblast of suckling rat, who were treated separately by drug serum containing high and low dose SJHYR and its disassembled prescriptions (Shengji formula and Huayu formula). The type I and III collagen contents in the fibroblasts were determined with immunocytochemical ABC method. RESULTS: Shengji formula could increase the levels of type I and III collagens in fibroblasts, which was significantly higher than those in model cells and control (P < 0.01). Huayu formula lowered them to the levels below those in model cells (P < 0.01), while in comparing the levels in the cells treated by high or low dose SJHYR with those in control, no significant difference was shown(P > 0.05). CONCLUSION: SJHYR might accelerate wound healing toward skin restoration through regulating the ratio of collagen type I and III, to adjust their metabolism. PMID- 12585108 TI - [Comparative study on regulation patterns of compound prescriptions for kidney tonifying and for blood circulation activating on T-cell apoptosis related gene expression in aged rats]. AB - OBJECTIVE: To investigate the regulation pattern of the two compound prescriptions for Kidney tonifying, Yougui Yin and Bushen Yishou capsule, in down regulating T-cell apoptosis gene expression in aged rats. METHODS: Expressions of T-cell apoptosis promoting and inhibiting genes, including Fas, FasL, Bcl-2, Bax, TNFR1 and TNFR2, as well as activity of cysteine proteinase in cascade connection, such as Caspase 8 and Caspase 3 were determined by TUNEL labeled flow cytometry and fluorescence real-time quantitative RT-PCR technique. The difference between old and young rats was compared, and the different regulation patterns of the two compound prescriptions for Kidney tonifying and their effects on Caspase activity were compared with those of compound prescription for blood circulation activating. RESULTS: The two compound prescriptions for Kidney tonifying could effectively lower T-cell over-apoptosis in old rats, down regulate FasL and TNFR1 gene transcription, and decrease the activity of Caspase 8 and Caspase 3, while the compound prescription for blood circulation activating showed insignificant effect on T-cell over-apoptosis. CONCLUSION: Kidney deficiency is closely related to the T-cell over-apoptosis. The T-cell over apoptosis in old rats could be effectively improved by the two compound prescription for Kidney tonifying through down-regulating the apoptosis promoting genes FasL and TN-FR1 transcription. That is the unique regulation pattern of Kidney tonifying principle to T-cell apoptosis related gene in old rats. PMID- 12585110 TI - [Clinical observation on large dosage of shenfu injection in supplementary treating liver cirrhosis complicated with esophageal varix rupture bleeding]. PMID- 12585109 TI - [Protective effect of hydrophilic Salvia monomer on liver ischemia/reperfusion injury induced by pro-inflammatory cytokines]. AB - OBJECTIVE: To observe the changes of tumor necrosis factor alpha (TNF alpha), interleukin 8 (IL-8) in liver ischemia/reperfusion injury and the protective effect of hydrophilic Salvia monomer MP-1 on them. METHODS: Hypothermic hypoxia reoxygenation model of human liver sinusoidal endothelial cell line and ischemia/reperfusion model of isolated rat liver were used. TNF alpha and IL-8 were measured with ELISA kits. Cell injury was excluded by trypan blue stain, sinusoidal endothelial cell function was assessed by hyaluronic acid uptake rate through RIA. Liver function was assayed by alanine transaminase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) release as well as output of bile flow. RESULTS: During hypoxia reoxygenation, sinusoidal endothelial cell injury, TNF alpha and IL-8 increased significantly in time-dependent manner, while sinusoidal cell function decreased. Cell injury was positively correlated to the released amount of TNF alpha (r = 0.949, P < 0.05) and IL-8 (r = 0.892, P < 0.05), respectively, the mortality could be reduced within 6 hrs by adding anti TNF alpha monoclonal antibody and increased by treating with recombinant human TNF alpha. Function of isolated rat liver lowered alone the increasing of low temperature ischemia/reperfusion time. MP-1 could markedly lower the mortality of endothelial cells and TNF alpha and IL-8 release, it also could alleviate ischemia/reperfusion injury to isolated rat liver. CONCLUSION: TNF alpha participated in liver ischemia/reperfusion injury directly, and MP-1 might alleviate the injury through inhibiting TNF alpha and IL-8. PMID- 12585111 TI - [Clinical observation on gianlietong suppository in treating chronic prostatitis]. PMID- 12585112 TI - [Controlled observation on electro-acupuncture combined with clomipramine in treating obsession]. PMID- 12585113 TI - [Prevention and treatment of prostatitis]. PMID- 12585114 TI - [Progress of study on effect of Chinese materia medica on angiogenesis]. PMID- 12585115 TI - [Study on the mechanism of Chinese herbal medicine in treating diabetic nephritis]. PMID- 12585116 TI - [Opinions on exploration of syndrome differentiation on eight cardinal principles]. PMID- 12585117 TI - [English translation of Chinese medicine]. PMID- 12585118 TI - [Experiences of international exchange of TCM with English language]. PMID- 12585119 TI - [Minute on national symposium on liver fibrosis]. PMID- 12585120 TI - [Inhibitory effects of hydrocortisone on human polymorphonuclear leukocyte adhesion to human synovial cell]. AB - AIM: To investigate the inhibitory effects of hydrocortisone on human polymorphonuclear leukocyte (PMN) adhesion to human synovial cell (HSC), and to explore its mechanism. METHODS: MTT colorimetry was used to determine the adhesion effect of PMN to HSC. Cell-ELISA and RT-PCR methods were used to determine the expression of adhesion molecular ICAM-1 and VCAM-1. EMSA method was also used to observe the activity of nucleus transcription factor-kappa B (NF kappa B). RESULTS: Hydrocortisone inhibited TNF-alpha (50 U.mL-1 for 12 hours) and IL-1 beta (50 U.mL-1 for 12 hours)-induced adhesion of PMN to HSC (IC50 2.05 x 10(-6) mol.L-1 and 2.13 x 10(-7) mol.L-1, respectively) in a concentration dependent manner. Adhesion molecular VCAM-1 and ICAM-1 protein and mRNA (rather than ICAM-1) expression in HSC induced by TNF-alpha (50 U.mL-1) were inhibited significantly by hydrocortisone at 1 x 10(-6)-10(-5) mol.L-1. The activity of NF kappa B was also extensively inhibited by hydrocortisone at 1 x 10(-6)-10(-5) mol.L-1. CONCLUSION: Hydrocortisone inhibited TNF-alpha-stimulated PMN-HSC adhesion, and expression of VCAM-1 by suppressing the activity of NF-kappa B. PMID- 12585121 TI - [Inhibitory effects of isorhapotigenin on IL-8 production and mRNA expression induced with TNF alpha in normal human synovial cells]. AB - AIM: To study the effects of isorhapotigenin (Iso) on interleukin-8 (IL-8) production and mRNA expression in normal human synovial cells (HSC) induced with TNF alpha. METHODS: IL-8 were assayed with RIA method. The mRNA expression of IL 8 was detected by RT-PCR method. RESULTS: It was shown that TNF alpha at concentrations of 0.05 to 0.5 U.mL-1 for 24 h significantly increased IL-8 production. The expression of IL-8 mRNA was also promoted by TNF alpha (0.25 U.mL 1) for 6 h. Iso at the concentrations of 1 x 10(-6) mol.L-1 to 1 x 10(-5) mol.L-1 showed inhibitory effects on IL-8 production induced with TNF alpha (0.25 U.mL 1). The further study indicated that Iso at the concentrations of 1 x 10(-6) mol.L-1 to 1 x 10(-5) mol.L-1 inhibited IL-8 mRNA expression in HSC induced with TNF alpha (0.25 U.mL-1). CONCLUSION: TNF alpha promoted IL-8 production and mRNA expression in HSC. Iso inhibited IL-8 production and mRNA expression induced by TNF alpha (0.25 U.mL-1). This might be one of the anti-inflammatory mechanisms of Iso. PMID- 12585123 TI - [Studied on docking of 5,6-diaryl-2,3-dihydro-1-pyrrolizinone derivatives with cyclooxygenase]. AB - AIM: To theoretically explore the mechanism of action of 5,6-diaryl-2, 3-dihydro 1-pyrrolizinone derivatives. METHODS: The interactions of the compound ZZ-122 with cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) were modeled by docking method. RESULTS: According to the binding pattern, intermolecular energy and capacity to form H-bond, it was easy for ZZ-122 to bind to COX-2 and not easy to COX-1. CONCLUSION: Compound ZZ-122 may be a selective COX-2 inhibitor, which has to be confirmed by experiment. PMID- 12585122 TI - [Possible mechanism of the protective effects of ginsenoside Rg1 on apoptosis in PC12 cells]. AB - AIM: To explore the possible mechanism of the protective effect of ginsenoside Rg1 on dopamine-induced apoptosis in PC12 cells. METHODS: Flow cytometric assay was performed to quantify the apoptotic cells and detect the positive percentage of Bcl-2 and Bax proteins in PC12 cells. The expression of bcl-2 and bax mRNA was identified by reverse transcription polymerase chain reaction (RT-PCR). The CPP32 activity was measured by fluorescent spectrofluorometer. RESULTS: Dopamine induced apoptosis in PC12 cells was inhibited by pretreatment with 10 mumol.L-1 Rg1. Simultaneously, the expression of Bcl-2 protein and mRNA were increased, and the Bax protein and mRNA were decreased. The activation of CPP32 was inhibited after pretreatment with Rg1. CONCLUSION: Rg1 protects against dopamine-induced apoptosis in PC12 cells and this effect might be attributed to its regulation of the ratio of Bcl-2 to Bax and its inhibition of the activation of CPP32. PMID- 12585124 TI - [Synthesis and antibacterial activity of 5-amino-6, 8-difluoro-1-(5-fluoro-2 pyridyl)-7-(3-methyl-1-piperazinyl)-1, 4-dihydro-4-oxo-3-quinolinecarboxylic acid and its analogues]. AB - AIM: To design and synthesize 1-(5-fluoro-2-pyridyl) quinolone derivatives, and to study their in vitro antibacterial activities. METHODS: Eight new compounds of 5-amino-6,8-difluoro-1-(5-fluoro-2-pyridyl)-7-(3-methyl-1-piperazinyl)-1, 4 dihydro-4-oxo-3-quinolinecarboxylic acid and its analogues were synthesized from ethyl 6-nitro-2, 3, 4, 5-terafluorobenzoylacetate and ethyl 3-methoxy-2, 4, 5 trifluorobenzoylacetate through 5 or 6 steps. RESULTS: Fifteen new compounds (1 15) were obtained including 8 desired compounds (8-15). The structures of the compounds were determined by 1HNMR, MS. CONCLUSION: In vitro antibacterial activities of the compounds (8-15) against Staphylococcus aureus-16, Escherichia coli-26 and Pseudomonas aeruginosa-17 were lower than control of ciprofloxacin. PMID- 12585125 TI - [Three dimensional quantitative structure-activity relationship of farnesyl protein transferase inhibitors]. AB - AIM: To build a three dimensional structure model that correlates the biological activities and the structures of a series of farnesyl protein transferase (FPT) inhibitors exemplified by the compound of 2, 3,4,5-tetrahydro-1-(1H-imidazol-4 ylmethyl)-4-(2-biphenylylcarbonyl)-1H-1, 4-benzodiazepine. METHODS AND RESULTS: Thirty-two FPT inhibitors with two types of scaffold were analyzed. Active conformations of which were studied using system search, a 3D-QSAR model were constructed using the method of comparative molecular field analysis (CoMFA). The resulting of cross-validated RCV2 = 0.602, non-cross-validated R2 = 0.958, SE = 0.270 and F = 124.5 indicate that the 3D-model possesses an ability to predict activities of new inhibitors. CONCLUSION: The information of CoMFA model offers an approach to designing new FPT inhibitors. PMID- 12585126 TI - [Study on the metabolism of psuedo-ginsenoside F11 in rats]. AB - AIM: To study the metabolic pathways of psuedo-ginsenoside F11 in rats. METHODS: By use of the prep-TLC, the metabolites were isolated. RESULTS: From the feces, three metabolites, psuedo-ginsenoside-RT5, ocotillol, and a new compound, F-3-1 (44) were isolated and identified by means of NMR evidence. But, such metabolites were found from neither the bile nor the urine of the rat. F-3-1 was identified as 6-O-alpha-L-rhamnopyranosyl-(1-2)-beta-D-glucopyranosyl-(20S,23S,24R)- dammar 20(24)-epoxy-3 beta, 6 alpha, 12 beta, 23,25-pentanol. CONCLUSION: Psuedo ginsenoside-F11 can not be metabolized in the rat liver, but can be metabolized in the large intestine of the rat. PMID- 12585127 TI - [Metabolic regulation of phenylethanoid glycosides from Herba cistanches in dogs' gastrointestine]. AB - AIM: To study the metabolic process of phenylethanoid glycosides (PhGs) in the gastrointestine of beagle dogs that were administered intragastrially process, and develop some new methods of biopharmacology on the effective position of traditional Chinese medicine. METHODS: High-performance liquid chromatography was used to purify constituents from faeces and analyze relative contents of the three main compounds in the gastrointestinal tract at different times and in the faeces of dogs. Every sample was collected, extracted with methanol and analyzed with integration. RESULTS: Four compounds, based on reference substances, were identified as echinacoside, acteoside, isoacteoside, and 2'-acetylacteoside from extraction of faces of dogs. Quantitative "with HPLC" analysis reveals that variation of ratios of the three main compounds is not distinct when moving in the gastrointestinal tract 7 h, that is quite different from those in faeces, in which the content of echinacoside fell from 48.0% to 16.0%, and acteoside increased from 11.0% to 34.7%. CONCLUSION: PhGs are mainly metabolized in large intestine. Among them, a portion of echinacoside is transformed into aceteoside. PMID- 12585128 TI - [The contents and their change of nucleosides from natural Cordyceps sinensis and cultured cordyceps mycelia]. AB - AIM: To compare the content of nucleosides from natural Cordyceps sinensis and cultured Cordyceps mycelia, and the effect of humidity and heat on the contents of nucleosides. METHODS: The contents of nucleosides were determined using high performance capillary electrophoresis (HPCE). Beckman P/ACE System 5010 apparatus equipped with a UV detector and a Beckman untreated fused-silica capillary (57 cm x 75 microns, 50 cm effective length) were used. Before sample injection, the capillary was rinsed with 1 mol.L-1 sodium hydroxide and running buffer for 5 minutes, respectively. 20 kV was applied during separation. Pressure injection was 586 kPa for 6 seconds, and the wavelength of detector was 254 nm. The running time was 20 minutes at 20 degrees C. The effect of humidity and heat on the content of nucleosides from natural Cordyceps sinensis and cultured Cordyceps mycelia was observed for 1, 3, 5 and 10 days at temperature 40 degrees C, and relative humidity 75%. RESULTS: The contents of nucleosides from cultured Cordyceps mycelia were higher than that of those from natural Cordyceps sinensis. The contents of nucleosides from freshly collected natural Cordyceps sinensis, were under the detection limit. The contents of nucleosides from natural Cordyceps sinensis were significantly increased by effect of humidity and heat, the phenomenon was not observed in cultured Cordyceps mycelia. CONCLUSION: There are difference in nucleosides content between natural Cordyceps sinensis and cultured Cordyceps mycelia. The nucleosides in natural Cordyceps sinensis may be derived from the degradation of macro-molecular nucleic acids. That means it is probably controversial that adenosine was used for quality control of natural Cordyceps sinensis. PMID- 12585129 TI - [Study on the phase II metabolites of phenoprolamine hydrochloride in rat bile by LC/DAD/MSD]. AB - AIM: To study the phase II metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. METHODS: DDPH was administered by i.p. to bile duct-cannulated rats. Bile samples were collected before drug administration and up to 12 h after drug administration. After being purified and enriched with C-18 SPE columns the rat bile samples were analyzed by LC/DAD/MSD to identify the peaks of phase II metabolites. The fractions of phase II metabolites were prepared by HPLC and treated with beta-glucuronidase, and then were purified and enriched with C-18 SPE columns and analyzed by LC/DAD/MSD. The corresponding reference standards of DDPH phase I metabolites were analyzed by LC/DAD/MSD under identical conditions. RESULTS: The peaks M7, M8 and M9 in the chromatograms of rat bile samples were the phase II metabolites of DDPH and the enzymatic hydrolysates of M7, M8 and M9 were 1-(2, 6-dimethyl-4-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M3), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M2) and 1-(2,6-dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethyl-amino)-propane (M1) respectively. CONCLUSION: beta-1-O-[3,5-dimethyl-4-[-2-methyl-2-(3,4 dimethoxy-phenylethylamino)- ethoxy]-phenyl]-glucuronic acid (M7, glucuronide of M3), beta-1-O-[2, 4-dimethyl-3-[2-methyl-2-(3, 4-dimethoxy-phenylethylamino) ethoxy]-phenyl]-glucuronic acid (M8, glucuronide of M2) and beta-1-O-[2-methoxy-4 [1-methyl-2-(2, 6-dimethylphenoxy)-ethylamino-ethyl]-phenyl]-glucuronic acid (M9, glucuronide of M1) were the phase II metabolites of DDPH in rat bile. PMID- 12585130 TI - [Effects of different phospholipids on the stabilities of doxorubicin liposomes in vitro and in vivo]. AB - AIM: To study the effects of egg phosphatidylcholine (EPC) and hydrogenated egg phosphatidylcholine (HEPC) on the leakage of doxorubicin liposome in vitro and the resident time in blood. METHODS: By the means of dialysis, the leakage of EPC (EPC-NL) and HEPC normal liposomes (HEPC-NL) in fetal cattle serum (FCS) was determined at 37 degrees C and in phosphate buffer solution (PBS) at 37 degrees C, 20 degrees C and 4 degrees C. Furthermore, the pharmacokinetics of EPC and HEPC sterically stabilized liposomes (EPC-SSL and HEPC-SSL) were studied by HPLC. RESULTS: The leakage of doxorubicin from HEPC-NL is slower than that of EPC-NL in FCS at 37 degrees C. But in PBS at all of three different temperatures, the results are completely reversed, i.e., the leakage of EPC-NL is slower than that of HEPC-NL. Further studies on pharmacokinetics showed that the mean residence time of HEPC-SSL in blood is 23.3 h, while that of EPC-SSL is 12.0 h, and the area under the curve (AUC) of concentration of HEPC-SSL is larger than that of EPC-SSL. CONCLUSION: HEPC-SSL is a better carrier in delivering the drugs to the extravascular sites than EPC-SSL. PMID- 12585132 TI - [Thermal decomposition kinetics of ribavirin and its stability]. AB - AIM: To study Thermal stabilities and thermal decomposition process of ribavirin and establish thermal decomposition kinetics equation. METHODS: Thermal weight loss curve was obtained by thermogravimetry balance. Thermal decomposition function mode was also determined by Achar differential method and Coats-Redfern integral method. RESULTS: Thermal decomposition kinetics parameters and kinetics compensation parameters were calculated from thermogravimetry-differential thermogravimetry data. Activation energy deduced by extrapolation under heating rate of 0 degree C.min-1 was 188.04 kJ.mol-1. Thermal decomposition kinetics function expression was d alpha/dt = Ae [formula: see text] (1 - alpha)2 and the mathematic expression of the kinetic compensation effect is found to be InA = 0.2264 Ea - 5.4458. CONCLUSION: Ribavirin has high thermal stability because of its high thermal decomposition activation energy. The thermal decomposition activation of capsule is little lower than that of material, which indicates no evident difference in thermal stability for capsules and material. PMID- 12585131 TI - [Structure characteristics of insulin liposomes]. AB - AIM: To study the molecular location and state of insulin within insulin-lipids. METHODS: Particle size and distribution of insulin-lipids was determined by using laser light scattering instrument, and transmission electron microscopy (TEM) was used to examine the morphology of insulin-lipids. HPLC, fluorescence scanning, digestion with trypsin and polyacrylamide gel electrophoresis was used to determine the molecular localization and state of insulin in the lipids. RESULTS: The mean particle size, polydispersity and sample quality were 218.3 nm, 0.073 and 7.2 respectively. The results of HPLC determination showed insulin exist in the insulin-lipids. There was no insulin emission spectrum in the insulin-lipids fluorescence spectrum; nearly no insulin digested in the test of insulin digestion by trypsin, and no insulin band in the insulin-lipids in figure of polyacrylamide gel electrophoresis. CONCLUSION: Insulin was encapsulated within lipids. PMID- 12585133 TI - [Optimization of the preparation of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine pharmacosomes using central composite design]. AB - AIM: To optimize the preparation of 3', 5'-dioctanoyl-5-fluoro-2'-deoxyuridine pharmacosomes (DO-FUdR-PS) by using central composite design. METHODS: DO-FUdR-PS was prepared by a thin-layer ultrasonication technique. The effects of drug phosphatidycholine ratio, pluronic F-68 concentration (%, w/v) and glycerol tristearate (%, w/v) concentration on the mean particle size, entrapment ratio (ER) and drug loading (DL) were investigated. A second-order polynomial equation was fitted to the data and the resulting model was used to predict the response in the optimal region. RESULTS: All the investigated response variables were found to be highly dependent on the formulation variables. Under the optimized conditions, the mean particle size, ER and DL of DO-FUdR-PS were 76 nm, 97.49% and 31.44%, respectively, which highly agreed with the predicted values. CONCLUSION: Central composite design was successfully used to optimize the preparation of DO-FUdR-PS. PMID- 12585134 TI - [Assignments of 1HNMR fingerprint of the root bark of Celastrus angulatus]. AB - AIM: To assign the 1HNMR finger-print of the root bark of Celastrus angulatus. METHODS: Silica gel column chromatography was used to separate the chemical constituents of CGE A of the root bark of C. angulatus. The characteristic signals of the 1HNMR finger-print were assigned after determining the structures of the compounds isolated from CGE A. RESULTS: 1HNMR finger-prints of the samples of C. angulatus collected from different regions showed highly characteristic features and reproducibility. Three compounds predominant in CGE A were isolated and their structures were determined by spectral analysis as: angulatin A (1), angulatin B (2) and angulatin C (3). CONCLUSION: Compound 3 is a new compound. Compound 2 was isolated from C. angulatus for the first time. The 1HNMR finger print of CGE A of the root bark of C. angulatus showed mainly the characteristic signals of the above three compounds and might be utilized for the original authentication of this plant. PMID- 12585135 TI - [A chemiluminescence flow sensor for vitamin B1]. AB - AIM: To determine vitamin B1 in pharmaceutical preparations by chemiluminescence (CL) flow sensor. METHODS: When 200 microL of Na3PO4 was passed through an anion exchange column, K3Fe(CN)6 was eluted from the resin and then mixed with the vitamin B1 stream containing NaOH to product CL, by the fast oxidation reaction between vitamin B1 and K3Fe(CN)6 in alkaline solution. RESULTS: The CL emission intensity was correlated with the vitamin B1 concentration in the range 1.0 x 10( 5) to 1.0 x 10(-3) mol.L-1 and the detection limit was 8.0 x 10(-6) mol.L-1 (3 sigma). A complete analysis, including sampling and washing, can be performed within 2 min with a relative standard deviation of less than 4.0%. CONCLUSION: The vitamin B1 flow sensor was stable for over 200 analyses and has been applied successfully to determination of vitamin B1 in pharmaceutical preparations. PMID- 12585136 TI - [Positron emission tomography imaging and drug development]. PMID- 12585138 TI - [The unite medicine in post genome era--prespect on the new frontier of complex sciences in 21 century]. PMID- 12585137 TI - Quality control for Chinese herbal drugs using DNA probe technology. PMID- 12585139 TI - [Stregthen the monitoring and studying on adverse reactions of Chinese herbal medicines]. PMID- 12585140 TI - [To accumulate experiences widely for reaching the goal through tortuous ways]. PMID- 12585141 TI - [Clinical observation on effect of zhenqi zhuanyin decoction combined with intrauterine insemination in treating spleen-kidney deficiency type patients of sterility with positive anti-sperm antibody]. AB - OBJECTIVE: To compare the effect of combined therapy of Zhenqi Zhuanyin Decoction (ZQZYD) and timely intrauterine insemination (IUI) with that of using ZQZYD and IUI separately in treating patients of sterility with positive anti-sperm antibody (AsAb). METHODS: One hundred and three patients were randomly divided into 3 groups, 34 patients in Group A treated with combined ZQZYD and IUI, 34 in Group B treated with ZQZYD alone and 35 in Group C treated with IUI alone. The negative conversion rate of AsAb, pregnancy rate in patients, pregnancy rate in AsAb converted patients, as well as changes of TCM Syndrome and T-lymphocyte subsets were observed. RESULTS: (1) The negative conversion rate of AsAb in the 3 groups was 76.47% (26/34), 82.35% (28/34) and 8.57% (3/35) respectively. (2) In Group A, the pregnancy rate was 41.18% (14/34), pregnancy rate in AsAb negative converted patients was 50.0% (13/26); in Group B, the respective criteria was 20.59% (7/34) and 21.43% (6/28) respectively; and in Group C, the pregnancy rate was 11.43% (4/35). The pregnancy rate in Group A was significantly higher than that in Group B or C (P < 0.01). (3) The effective rate on TCM Syndrome in Group A and B was 88.23% and 91.18% respectively (P > 0.05). (4) In Group A and B, CD4 decreased, CD8 increased and CD4/CD8 ratio decreased after treatment, which were significantly different from those before treatment and in Group C after treatment (P < 0.05). CONCLUSION: Combined therapy of ZQZYD and IUAI shows a better effect in treating infertility with positive AsAb than that by ZQXYD or IUI alone. PMID- 12585142 TI - [Effect of xiaoyu tablet on blood flow parameters and morphology of carotid artery in atherosclerotic patients]. AB - OBJECTIVE: To study the effect of Xiaoyu Tablet (XYT) on blood flow parameters and morphology of carotid artery in atherosclerotic patients. METHODS: Using color Doppler ultrasonographic technique to examine the blood flow parameters and intimal thickness of carotid artery in 20 patients of carotid atherosclerosis after 24 weeks treatment of XYT, and compared with those in 10 patients treated with gastrodine lipid-lowering tablet. RESULTS: After 24 weeks treatment, blood flow parameters of carotid artery were obviously improved and intimal thickness of common carotid arteries in both side was markedly decreased. XYT showed an effect better than that of gastrodine lipid-lowering tablet. CONCLUSION: XYT is effective in increasing blood flow of cervical and cerebral arteries. PMID- 12585143 TI - [Effect of wuzi yanzong Pill on mitochondrial DNA deletion and respiratory chain enzyme complex activity in peripheral leukocyte of aged male with kidney deficiency syndrome]. AB - OBJECTIVE: To investigate the effect of Wuzi Yanzong Pill (WZYZP) on mitochondrial DNA (mtDNA) deletion and respiratory chain enzyme complex (RCZC) in peripheral blood leukocyte of aged male with Kidney Deficiency Syndrome (KDS). METHODS: Single-blinded study was conducted in 38 aged male with KDS, who were randomly divided into 2 groups treated with WZYZP and placebo respectively for 3 months. Levels of mtDNA deletion and RCZC were determined by polymerase chain reaction (PCR) and enzyme kinetics technique respectively. RESULTS: WZYZP could reduce the mtDNA deletion and raise the activity of mitochondrial RCZC I, IV in peripheral blood leukocyte of aged male with KDS (P < 0.05, P < 0.01). CONCLUSION: WZYZP has protective effect on mtDNA from oxidative damage in leukocyte of aged male with KDS. PMID- 12585145 TI - [Influence of needling at yin-yang meridian points on cerebral glucose metabolism]. AB - OBJECTIVE: To study the different effect of needling at different meridian points (MP) on different cerebral functional regions by using positron emission tomography and cerebral function tracer 18F-fluorodeoxy glucose to observe the effect of electro-acupuncture (EA) stimulation at Yin MP or Yang MP on local cerebral glucose metabolism in 30 healthy volunteers. METHODS: The thirty subjects were randomly divided into 2 groups, the 12 subjects in the Yang MP group treated by needling at Hegu (LI4), Quchi (LI11), Zusanli (St36) and Shangjuxu (St37), the 18 subjects in the Yin MP group treated by needling at Neiguan (Pc6), Chize (Lu5), Sanyinjiao (Sp6) and Yinlingquan (Sp9). All the needling was conducted at one side of body. RESULTS: After needling at Yang MP, the cerebral glucose metabolism was elevated in bilateral cross region of frontal lobe and temporal lobe, parietal lobe, thalamus and basal ganglia, and in contralateral cerebellum and hippocampus. After needling at Yin MP, the cerebral glucose metabolism was lowered in bilateral cross region of frontal and temporal lobe, contralateral temporal lobe, cerebellum and thalamus, and was elevated in homolateral hippocampus and caudate nucleus. CONCLUSION: Under physiological condition, the regulatory effects of acupuncture on central nervous system would be different when needling at different MP. PMID- 12585144 TI - [Influence of combined therapy of guben yiliu III, moxibustion and chemotherapy on immune function and blood coagulation mechanism in patients with mid-late stage malignant tumor]. AB - OBJECTIVE: To observe the supplementary effect of moxibustion and Guben Yiliu III (GBYL), a Chinese herbal compound preparation, in combination with chemotherapy. METHODS: Eighty-one patients of mid-late stage malignant tumor were randomly divided into 3 groups: 16 in Group A treated with chemotherapy and placebo; 35 in Group B treated with chemotherapy and GBYL and 30 in Group C treated with chemotherapy and GBYL plus moxibustion. The short-term effect of treatment, changes of blood picture, cell mediated immune function and blood coagulation in patients were observed. RESULTS: After chemotherapy, the lymphocyte count was significantly lowered in Group A and B (P < 0.01), but not in Group C (P > 0.05); lymphocyte subset T3 raised significantly in Group B; the average level of T lymphocyte subsets was reduced in Group A while it increased in the other two groups; and a bi-directional regulation on plasma fibrinogen concentration was shown in Group C (P < 0.05). CONCLUSION: Moxibustion prevented dropping of lymphocyte count caused by chemotherapy. Combination of GBYL and moxibustion could prevent the lowering of T-lymphocyte subsets caused by chemotherapy, and moxibustion could regulate bi-directionally the patients' abnormality in part of blood coagulation mechanism. PMID- 12585146 TI - [Clinical study on effect of yufeng capsule on premonitory symptoms of apoplexy]. AB - OBJECTIVE: To observe the clinical effect of Yufeng Capsule (YFC), a Chinese herbal preparation with function of clearing Heat-toxin in treating premonitory symptoms of apoplexy in middle and old aged patients. METHODS: One hundred and fourteen patients with premonitory symptoms of apoplexy were treated with YFC and compared with 57 patients treated with Venoruton for control. The changes of clinical symptoms, anti-oxidation capability, blood lipid, blood sugar and hemorrheological parameters were observed. RESULTS: (1) The therapeutic effect in the YFC group was cured in 42 cases (36.84%), markedly effective in 38 (33.33%), effective in 28 (24.58%) and the total effective rate was 94.74%, while in the control group, the corresponding numbers were 9 (15.79%), 8(14.04%), 20(35.09%) and 64.91% respectively, the difference between the two groups in cure rate, markedly effective rate and total effective rate was significant (P < 0.05); (2) The whole blood viscosity, plasma viscosity, hemagglutination index and red blood cell deformity index in the YFC group were significantly improved after treatment (P < 0.01) and the improvement was better than those in the control group (P < 0.05 or P < 0.01); (3) Levels of serum malonyldialdehyde (MDA), superoxide dismutase (SOD), blood lipids and glucose were markedly improved in the YFC group (P < 0.05) after treatment, and showed a significant difference to those in the control group (P < 0.05). CONCLUSION: YFC has obvious effect in treating premonitory symptoms of apoplexy patients, it could markedly improve the hemorrheologic parameters, regulate blood lipids and blood glucose metabolism, and strengthen the anti-oxidation capability of patients. PMID- 12585147 TI - [Determining blood parameters FT3, FT4, T, E2 and cortisol to explore nature of cold syndrome and heat syndrome]. AB - OBJECTIVE: To explore the relationship between the levels of free triiodothyronine (FT3), free thyroxine (FT4), cortisol (CO), testosterone (T), serum estradiol (E2) and Cold Syndrome and Heat Syndrome in traditional Chinese medicine. METHODS: The blood levels of FT3, FT4, T, E2, CO in groups of sthenic Heat (SH), sthenic Cold (SC), asthenic Heat (AH) and asthenic Cold (AC) and the healthy control group were measured. The number of cases in each group was 50. RESULTS: (1) FT3 and FT4 were higher in Heat Syndrome than that in Cold Syndrome in the order of AH > SH > control > SC > AC, it revealed that the basal metabolic rate and oxygen consumption in Heat Syndrome were increased, while it in Cold Syndrome, it declined. (2) CO was higher in SH, SC than that in AH and AC, the order was SC > SH > control > AH > AC, it revealed that CO was one of the factors correlated to asthenic and sthenic syndrome, but not relevant to Heat Syndrome and Cold Syndrome. (3) T was higher in Heat Syndrome than that in Cold Syndrome, but E2 showed the opposite, suggesting that T was one of the factors for forming Heat Syndrome, and E2 for Cold Syndrome. CONCLUSION: The levels of FT3, FT4, T, E2 are basis of pathophysiology related with forming HS and CS. PMID- 12585149 TI - [Effect of wenjingtong composita on blood glucose and advanced glycosylation end products in sciatic nerve of streptozotocin-induced diabetic rats]. AB - OBJECTIVE: To investigate the effect of Wenjingtong Composita (WJTC) on blood glucose and advanced glycosylation end products (AGEs) in sciatic nerve of streptozotocin (STZ)-induced diabetic rats. METHODS: STZ-induced diabetic rats were randomized to WJTC prevention group and WJTC treatment group. The levels of blood glucose and AGEs in sciatic nerve of the animals were checked after 12 weeks treatment and compared with that of aminoguanidine (AG) treatment group. RESULTS: Blood glucose level in the WJTC prevention and treatment group, and AGEs in sciatic nerve of the WJTC prevention group and the AG group were lower than those of the non-treated group (P < 0.01). Blood glucose level in the AG group was higher than that in the WJTC prevention group (P < 0.05), but was not significantly different from that in the non-treated group (P > 0.05). CONCLUSION: WJTC might prevent diabetic peripheral neuropathy by decreasing blood glucose and inhibiting AGEs formation in sciatic nerve in STZ-induced diabetic rats. PMID- 12585148 TI - [Observation on effect of danxiaoling in supplementary treatment of intrahepatic cholestasis in pregnancy]. AB - OBJECTIVE: To evaluate the curative effect of Danxiaoling Pill (DXLP), a Chinese herbal preparation, in treating intrahepatic cholestasis in pregnancy (ICP). METHODS: Fifty-eight cases of ICP were divided randomly into two groups and treated by DXLP and Composite Yiganling as control respectively with the other identical conventional treatment. The changes of clinical symptoms, related laboratory parameters after treatment and the condition of labor were observed. RESULTS: The total effective rate in both groups was 100%, but the markedly effective rate in the DXLP treated group was higher than that in control group (P < 0.01). Levels of blood cholyglycine acid (CGA), alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin, total cholesterol and low density lipoprotein were all decreased in both groups after treatment, but DXL showed a better efficacy in decreasing CGA, ALT and AST than that of Yiganling. Moreover, the amniotic fluid meconium contaminated rate, premature delivery occurrence in the DXLP group were lower than those in the control group, while the weight of newborn baby was higher in the former than in the latter. CONCLUSION: DXLP could effectively lower the serum bile acid and improve liver function in treating ICP. PMID- 12585151 TI - [Experimental study on liver protective effect of Artemisia rupestris extract]. AB - OBJECTIVE: To explore the protective effect of Artemisia rupestris extract (ARE) on chemical liver injury and immunological hepatitis. METHODS: Mice model of immunological hepatitis was established by lipopolysaccharide plus Bacillus Calmette-Guerin vaccine (BCG), and the models of liver injury was induced by D galactosamine or carbon tetrachloride (CCl4) in mice and rats. The effects of ARE in preventing and treating immunological hepatitis and protecting liver injury were observed. RESULTS: ARE significantly lowered the serum alanine transaminase (ALT) in immunological hepatitis (P < 0.05), CCl4 induced liver injury model mice and rats (P < 0.01), and in D-galactosamine induced liver injury mice (P < 0.05). CONCLUSION: ARE has significantly protective effect against chemical liver injury and could treat immunological hepatitis, but with no effect in preventing immunological hepatitis. These might be the partial pharmacologic basis for using ARE in treating hepatitis. PMID- 12585150 TI - [Effect of Erigeron breviscapus injection on ventricular and vascular remodeling in spontaneous hypertension rats]. AB - OBJECTIVE: To observe the ventricular and vascular remodeling reversal effect of Erigeron breviscapus injection (EBI), a protein kinase C inhibitor, in spontaneous hypertension rats (SHR). METHODS: Twenty-four SHR were divided into 4 groups, they were treated respectively with EBI, Fosinopril, Enalapril and normal saline 10 mg/kg per day by intraperitoneal injection for 8 weeks. Systolic blood pressure (SBP), ventricular weight index (VWI) and protein kinase C (PKC) activity in myocardial cells were determined, ultrastructural changes of heart and vessel were observed by polaroscope and transmission electron microscope, and the area and content of myocardial interstitial collagen (MIC) were determined by image analyzer system. RESULTS: The left ventricular hypertrophy was regressed to certain degree after EBI, Fosinopril and Enalapril treatment, but no significant change in heart rate and right VWI was found. Fosinopril and Enalapril were superior to EBI in lowering SBP and left VWI, and EBI was more obvious in improving myocardial ultrastructure such as hypertrophy and degeneration. All the 3 drugs could improve the MIC and vascular remodeling, the MIC area, content and collagen volume fraction in the EBI group were lowered after treatment, as compared with those in the control group, but comparison between the three groups showed no significant difference. The 3 drugs could reduce the PKC activity in myocardial cell membrane, and EBI showed the effect more significant than that of the other two (P < 0.05). CONCLUSION: EBI could reverse the myocardial, interstitial and vascular remodeling, improve the rigidness of cardiac muscle, thus, has protective effect on heart. PMID- 12585152 TI - [Effect of jiangu erxian pill on proliferation and cell cycle of human osteoblast]. AB - OBJECTIVE: To further explore the effect of Jiangu Erxian Pill (JGEXP) on proliferation and cell cycle of human osteoblast on the basis of previous clinical and experimental studies. METHODS: Human primary osteoblast were isolated and cultured. The cell proliferation was tested by 3H-thymine incorporation and methyl thiazolyl tetrazolium (MMT) method and the cell cycle was determined by flow cytometry technique. RESULTS: In the medium and high dosage JGEXP groups, the cell proliferation rate and index, and percentage of diploid synthesis phase (S phase) cells were significantly higher than those in the blank control group (P < 0.01 or P < 0.05), and similar to those in the estrogen group; and the cell apoptosis rate and percentage of G0-G1 stage cells were lower than those in the blank control group (P < 0.01 and P < 0.05). CONCLUSION: JGEXP could effectively promote the cell proliferation and differentiation, and prevent the cell apoptosis of osteoblast in vitro. PMID- 12585153 TI - [Effect of electro-acupuncture on pathology, water content in brain tissue and neurologic function defect scoring in rat model of idiopathic cerebral hemorrhage]. PMID- 12585154 TI - [Observation on short-term effects of haizhu gigan capsule in treating chronic hepatitis B]. PMID- 12585155 TI - [Comparative clinical observation on effects of shensu injection and vitamin C in treating acute viral myocarditis]. PMID- 12585156 TI - [Clinical observation on treatment of diabetes mellitus type 2 by medication with both Chinese and Western drugs]. PMID- 12585157 TI - [Query on the cause of "belgium nephropathy" (Chinese herb nephropathy) and the speculation on carcinogenesis of Chinese herbs]. PMID- 12585158 TI - [Application of laser scanner co-focal microscopy in researches of traditional Chinese medicine]. PMID- 12585159 TI - [The eventual application of fuzzy mathematics in traditional Chinese medicine]. PMID- 12585160 TI - [Progression of study in ovulation promotion of TCM]. PMID- 12585161 TI - [The unit medicine in post-era--prospect on the new frontier of complex science in the 21 century]. PMID- 12585162 TI - [Evidence-based medicine and integration of traditional Chinese, and Western medicine]. PMID- 12585163 TI - [Effect of gandou tablet I on neurological electrophysiology in patients of hepatolenticular degeneration]. AB - OBJECTIVE: To observe the influence of Gandou Tablet I (GD I) on neurologic electrophysiology in patients of hepatolenticular degeneration (HLD). METHODS: Vigilance-controlled electroencephalogram (VEEG), electromyogram (EMG) and brain stem auditory evoked potential (BAEP) in 32 cases of HLD were examined before GD I treatment, and reexamined after 4 weeks of treatment in those with abnormal neurologic electrophysiology. RESULTS: At the same time of clinical symptom improvement after treatment, the abnormal VEEG, EMG and BAEP also improved with the effective rate of 71.43%, 70% and 66.67% respectively, their constitutional ratio were different insignificantly, P > 0.05. The peak latency (PL) of III, V waves and interpeak latency (IPL) of I-III, III-V and I-V waves in patients before treatment were all greatly prolonged, compared with those in the healthy control group (P < 0.05 or 0.01), the IPL of III-V and I-V waves were markedly improved after treatment. CONCLUSION: Neurologic electrophysiologic examination was valuable for complementary diagnosis and therapeutic effect evaluation of HLD. GD I could improve the abnormal neurologic electrophysiologic parameters. PMID- 12585164 TI - [Clinical and experimental study on jiangzhi tiaoya granule in treating essential hypertension and protecting function of vascular endothelium]. AB - OBJECTIVE: To observe the therapeutic effect of Jiangzhi Tiaoya Granule (JZTYG) in treating essential hypertension and its protection on function of vascular endothelial cells (VEC). METHODS: Fifty-nine patients of essential hypertension divided into two groups were treated with JZTYG (the treated group) and Jinjia Yixintong (the control group) respectively. The changes of symptoms, signs, blood pressure, heart rate were observed and the levels of endothelin (ET), calcitonin gene related peptide (CGRP) content were determined by radioimmunoassay (RIA). RESULTS: The total effective rates of JZTYG in lowering blood pressure and improving symptoms were both 90.0%, markedly effective rate in lowering blood pressure and improving symptoms was 36.7% and 60.0% respectively. The symptom improved in the treated group was better than that in the control group (P < 0.05). It also could reduce the plasma ET level (P < 0.05) and ET/CGRP ratio (P < 0.01), and increase the CGRP level (P < 0.05). CONCLUSION: JZTYG has a promising clinical therapeutic effect in treating essential hypertension and is able to protect the VEC function. PMID- 12585165 TI - [Study on interventing effect of puerarin on insulin resistance in patients with coronary heart disease]. AB - OBJECTIVE: To explore the effect of puerarin in improving the insulin resistance (IR) and its closely related abnormal lipid and fibrinolytic activity in patients with coronary heart disease (CHD). METHODS: Seventy-six patients with CHD were randomly divided into two groups, 40 in the puerarin group and 36 in the routine treated group. Puerarin 500 mg was given to the former in addition to routine therapy by adding to 250 ml of normal saline for intravenous dripping once a day with a therapeutic course of 3 weeks. The changes of fasting blood glucose (FBG), fasting plasma insulin (FINS), plasma total cholesterol (TC), triglyceride (TG), low and high density lipoprotein cholesterol (LDL-C & HDL-C) and plasminogen activator inhibitor-1 (PAI-1) activity were measured before and after treatment, and the insulin sensitivity index (ISI) calculated. At the same time, tissue plasminogen activator (tPA) activity before and during venous occlusion test (VOT) was tested. Besides, 30 healthy subjects were taken as control. RESULTS: In CHD patients, FINS, TC, TG, LDL-C and PAI-1 levels were higher and ISI, HDL-C and tPA before and during VOT were lower than those in the healthy controls. FINS and ISI correlated well with lipids and fibrinolytic abnormality. After puerarin treatment, FINS level lowered and ISI increased significantly (P < 0.01), while comparing with the routine group, TC, TG, LDL-C and PAI-1 were lower but HDL-C and tPA activity before and during VOT were higher in the puerarin group (P < 0.05, P < 0.01). Correlation analysis showed that FINS was positively correlated with TC, TG, LDL-C and PAI-1 and negatively correlated with HDL-C, tPA before and during VOT; ISI was negatively correlated with TC, TG, LDL-C and PAI-1 and positively correlated with HDL-C and tPA before and during VOT in the puerarin group. CONCLUSION: Puerarin could improve the IR, IR related lipid and fibrinolytic activity abnormality in CHD patients. PMID- 12585166 TI - [Effect of bushen huayu composite on secretion of lymphocytic interleukin-2 and interleukin-2 receptor expression in the aged]. AB - OBJECTIVE: To study the effect of Bushen Huayu Composite (BSHY) on interleukin-2 (IL-2) secretion and IL-2 receptor expression in the aged. METHODS: IL-2 was examined by methyl thiazolyl tetrazolium (MTT) method and IL-2 dependent cellular stains, IL-2 receptor expression was examined by FACS-indirect immunofluorescence. RESULTS: Aging could result in the decrease in lymphocytic IL 2 secretion and IL-2 receptor expression (P < 0.01, P < 0.05), which could be improved by BSHY (P < 0.01, P < 0.05). CONCLUSION: BSHY has significant up regulatory effect on immune function of lymphocytes in the aged. PMID- 12585167 TI - [Study on effect of Salvia injection in treating primary nephrotic syndrome and on endothelin and serum interleukin-2 receptor in children]. AB - OBJECTIVE: To explore the effect and mechanism of Salvia Injection (SI) in treating primary nephrotic syndrome (PNS) in children. METHODS: Forty-four PNS children were randomly divided into conventional steroid treated group (20 cases) and conventional plus SI intervention treated group (24 cases), the levels of serum endothelin (ET), soluable interleukin-2 receptor (sIL-2R) were observed. RESULTS: Before treatment, plasma ET and sIL-2R in PNS children were higher than those in healthy children significantly (P < 0.01). After treatment, the ET and sIL-2R levels were all obviously improved in both treated groups (P < 0.05) and the improvement in the conventional plus SI intervention treated group was more obvious, the difference between the two treated groups after treatment was significant (P < 0.05). CONCLUSION: Conventional treatment supplemented with SI could more effectively improve the levels of plasma ET and SIL-2R in treating PNS children, and hence alleviate the damage of renal tissue. PMID- 12585168 TI - [Clinical observation on colquhounia root tablet in treating lipid metabolism disturbance secondary to nephrotic syndrome]. AB - OBJECTIVE: To study the effect of Colquhounia root tablet (CRT) in treating nephrotic syndrome with sequential lipid metabolism disorder (NS-LMD). METHODS: The 96 patients with NS-LMD were randomly divided into two groups, the 60 cases in the treated group treated with CRT and the 36 cases in the control group treated with hormone or cytotoxic medicine. The curative effect and the related indexes were determined before and after treatment. RESULTS: After six months treatment, the general effective rate in the treated group was 88. 33%, which was markedly higher than that in the control group (69.44%, P < 0.05). The levels of the treated group in ameliorating lipid metabolism disorder and renal dysfunction were also higher than those in the control group (P < 0.05). CONCLUSION: CRT could improve NS-LMD, improve renal function, eliminate urinary protein and increase plasma albumin. It is highly effective with low toxicity and safe. PMID- 12585169 TI - [Exploration on essence of spleen-kidney deficiency in middle-aged patients]. AB - OBJECTIVE: To explore the essence of Spleen-Kidney Deficiency in middle-aged patients. METHODS: Investigation was carried out in 773 cases of 50-69 years old to assay 34 parameters in them, including blood lipid, oxidation and anti oxidation related substance, sex hormone, liver and renal function, immune function, blood routine, blood pressure and lung vital capacity, etc. RESULTS: Spleen Deficiency Syndrome is closely related with lipid metabolism disorder; Kidney-Qi Deficiency Syndrome is closely related with hypoimmune function; Kidney Yin Deficiency Syndrome is closely related with lipid metabolism disorder and hypertension, and Kidney Yang Deficiency is closely related with weakness of anti oxidation capacity, hypoimmunity, the internal environment disorder of sex hormone and aging, manifested as multiple functions abating. CONCLUSION: Spleen Deficiency, Kidney-Qi Deficiency and Kidney Yang Deficiency are different layers of a gradually developed and aggravated pathological process, but Kidney-Yin Deficiency could not be listed into this layer. PMID- 12585170 TI - [Clinical and experimental study on treatment of acute cerebral infarction by zhongfeng no. II oral liquid]. AB - OBJECTIVE: To explore the effect and mechanism of Zhongfeng No. II Oral Liquid (ZF II) in treating acute cerebral infarction (CI). METHODS: Randomly controlled clinical study was conducted in 160 patients with CI, they were treated with ZF II plus western medicine (treated group) and western medicine alone (control group) respectively. The animal model of CI was made by unilaterally common carotid artery ligation in 45 Mongolian gerbil, and they were divided into the model group, the normal saline group and ZF II group. RESULTS: ZF II could improve the TCM Syndrome, neuro-function and cerebral blood circulation in CI patients. Experimental study showed that ZF II could significantly inhibit the release of glutamic acid in 6 hrs (P < 0.05), maintain the level of superoxide dismutase (SOD) and glutathione, and scavenge the free radicals effectively. CONCLUSION: ZF II could improve the function of brain tissue, and is worth developing and applying in treating CI. PMID- 12585171 TI - [Effect of shenmai injection and aminophylline on small airway smooth muscle cell apoptosis and related gene expression in rats with emphysema]. AB - OBJECTIVE: To investigate the effect of Shenmai Injection (SMI) and aminophylline on small airway smooth muscle cell (SASMC) apoptosis and the Fas/FasL expression in the papain induced emphysema model rats. METHODS: Emphysema model in rat was established by a single intratracheal instillation of papain. Apoptosis and Fas/FasL expression of SASMC were examined by immunohistochemical SABC and TUNEL assay at 1, 3, 5, 7, 15 and 30 days after modelling, and the effect of SMI and aminophylline on them were observed. RESULTS: Fas, FasL expressions in normal SASMC were very low with a positive rate of (2.31 +/- 0.05)% and (1.28 +/- 0.47)% respectively. After papain instillation, the positive rates increased along with the prolonging of instillation time. SMI showed an inhibition on SASMC Fas and FasL expression but aminophylline didn't show. SASMC apoptosis was very low in normal rats with a rate of (0.87 +/- 0.32)%, it also raised after papain instillation and increased progressively along with the instillation time. SMI treatment could lower the apoptosis rate but aminophylline couldn't. CONCLUSION: Fas and FasL participated the SASMC apoptosis modulation in emphysema formation. SMI shows a definite treatment effect on emphysema by influencing the Fas and FasL protein expression and reducing SASMC apoptosis through inhibiting the release of inflammatory mediator. PMID- 12585172 TI - [Effect of bushen yanggan recipe on nigrostriatal function in parkinsonian model rats after long-term levodopa treatment]. AB - OBJECTIVE: To investigate the effect of Bushen Yanggan Recipe (BSYGR) on the function and morphology of nigrostriatal system in Parkinsonian model rats with long-term levodopa treatment. METHODS: Unilateral Parkinsonian rat models were established by injecting 6-hydroxydopamine (6-OHDA) into the substantia nigra pars compacta (SNpc) and ventral segmental area (VTA). Animals were randomly divided into four groups, the sham control group, model control group, levodopa group and levodopa plus BSYGR group. The content of striatal dopa (DA), digydroxy phenyl acetic acid (DOPAC) and homovanilic acid (HVA) or the THmRNA expression level in the midbrain were measured. RESULTS: (1) Levels of striatal DA, DOPAC, HVA, DOPAC/DA, HVA/DA decreased in the model control group by about 90% as compared with those in sham control group (P < 0.05). These parameters in the levodopa group were higher than those in the sham control group, while in the levodopa plus BSYGR group, they were lower than those in the levodopa group (P < 0.01), approaching the levels in the sham control group (P > 0.05). (2) Striatal TH activity in the model group was lower than that in the sham control group significantly, but higher than that in the levodopa group, while in the levodopa plus BSYGR group, it showed a level obviously higher than that in the levodopa group (P < 0.05). (3) Levodopa plus BSYGR group had a higher midbrain THmRNA expression level than that in the levodopa group. CONCLUSION: BSYGR could effectively reduce the side effects resulting from the long-term treatment of levodopa. PMID- 12585173 TI - [Effect of Panax notoginseng saponins on human kidney fibroblast]. AB - OBJECTIVE: To study the effect of Panax notoginseng saponins (PNS) on cell proliferation, type I collagen secretion and integrin beta 1 expression in human kidney fibroblast (KFB). METHODS: KFB were cultured and stimulated by PNS in vitro. The cell proliferation, type I collagen secretion and integrin beta 1 expression in KFB were measured by methyl thiazolyl tetrazolium (MTT), enzyme linked immunoabsorbent assay (ELISA) and flowcytometry respectively. RESULTS: Within its optimal concentration range and effective time range, PNS could obviously inhibit the proliferation, type I collagen secretion and integrin beta 1 expression (all P < 0.05) in human KFB. CONCLUSION: PNS would possibly be an effective drug for renal interstitial fibrosis prevention and treatment. PMID- 12585174 TI - [Effect of bushen yizhi recipe on neurotransmitter release in A beta segment neurotoxin induced NG108-15 cellular model of Alzheimer disease]. AB - OBJECTIVE: To observe the effect of Bushen Yizhi Recipe (BSYZR) on neurotransmitter release in A beta segment neurotoxin induced NG108-15 cellular model of Alzheimer disease (AD). METHODS: The levels of choline acetyltransferase (ChAT) activity, synapsin and functional synapse formation rate in the cellular model treated with BSYZR containing serum were determined by Western blot analysis, immunoradiometric assay and electrophysiologic technique. RESULTS: BSYZR containing serum treatment could cause increase of ChAT activity and synapsin level in model cells, as compared with those in normal control model cells treated with non-drug containing serum, it also could regulate the release capacity of transmitter and raise the functional synapse formation. CONCLUSION: BSYZR could reduce the reaction of cell to A beta neurotoxin, indicating that it could be antagonistic to the pathological development of AD by means of raising the neurotransmitter release capacity. PMID- 12585175 TI - [Experimental study on effect of Angelica polysaccharide in inhibitory proliferation and inducing differentiation of K562 cells]. AB - OBJECTIVE: To investigate the application value of Angelica polysaccharide (APS) on proliferation and differentiation of human erythroleukemia K562 cells. METHODS: The effect of APS in inhibitory proliferation and inducing differentiation of human erythroleukemia K562 cells was studied by modern experimental hematologic techniques such as cell counting and culture, flowcytometry, morphology, cytochemistry and cell differential immune phenotyping. RESULTS: APS could significantly inhibit the proliferation of K562 cells in vitro and prevent the cell from entering the active proliferative phase (P < 0.05). After being induced by APS, the differentiation of K562 cells to erythrocyte series and granulo-monocyte series increased, positive rate of benzidine, glycogen and peroxidase stain elevated, and cell surface differential antigen CD15 expression promoted significantly (P < 0.05), while C-MYC expression of K562 cells induced by APS induction lowered significantly (P < 0.05). CONCLUSION: APS could not only inhibit the proliferation of K562 cells in vitro, but also induce the differentiation of K562 cells toward erythrocyte and granulocyte series. It may be a natural inducer with promising prospect of development and application. PMID- 12585176 TI - [Systematic evaluation of random controlled trial on treatment of chronic hepatitis B by Chinese herbal medicine]. PMID- 12585177 TI - [Observation on effects of qige tongye decoction combined with chemotherapy in treating esophageal carcinoma]. PMID- 12585178 TI - [Influence of moxibustion with small moxa-cone on the activity ptyalin and the contents of medium molecules in plasma to patients with diarrhea due to spleen deficiency]. PMID- 12585179 TI - [Progression of study on treatment of acute myocardial infarction with thrombolysis therapy of integrated tradition Chinese and Western medicine]. PMID- 12585180 TI - [Study of TCM syndrome]. PMID- 12585181 TI - [A summary of academic thought on integration of traditional Chinese and Western medicine]. PMID- 12585182 TI - [Clinical observation on effect of Chinese herbal medicine plus human chorionic gonadotropin and progesterone in treating anticardiolipin antibody-positive early recurrent spontaneous abortion]. AB - OBJECTIVE: To find a method without corticosteroids, aspirin or heparin for treatment of anticardiolipin antibody-positive early recurrent spontaneous abortion (AARSA). METHODS: Twenty-three patients of AARSA in the treated group were treated with Chinese herbal medicine (CHM) plus human chorionic gonadotropin and progesterone, and 18 patiens in the control group were treated with multi vitamin only. The change of anticardiolipin antibody was determined by enzyme linked immunoabsorbent assay (ELISA). RESULTS: After treatment, anticardiolipin antibody negative converted in 20 cases (86.9%) of the treated group. The cure rate of abortion in the treated group was 82.6% (19/23), which was raised to 95% (19/20) in those patients with antibody negative conversion, while in the control group, it was 16.7% (3/18) merely, comparison between the two groups in cure rate showed significant difference (P < 0.01). CONCLUSION: CHM plus human chorionic gonadotropin and progesterone could cure AARSA effectively. PMID- 12585183 TI - [Observation on treatment of ectopic pregnancy by combination therapy of Chinese herbal medicine with mifepristone or methotrexate]. AB - OBJECTIVE: To find a simpler, effectiver with less side-effect method for treatment of ectopic pregnancy. METHODS: One hundred and fourteen patients of ectopic pregnancy were divided into two groups, the 56 patients in Group A were treated with Mifepristone and Chinese herbal medicine (CHM), and 58 patients in Group B were treated with Methotrexate and CHM. The therapeutic effect in both groups was observed and compared. RESULTS: The rate of success in Group A was 82.14% and in Group B 70.69%, comparison between them showed no significance (P > 0.05). Time for successful treatment was 14.54 +/- 5.87 days in Group A and 9.00 +/- 5.09 days in Group B, comparison of them showed significant difference (P < 0.01). Insignificance revealed in comparison between the occurrence of side effects for the groups, such as nausea, vomiting, diarrhea, liver dysfunction and lowering of white blood cells, but significance was shown in re-menstruation time (43.43 +/- 1.77 days vs 38.38 +/- 1.13 days, P < 0.05). CONCLUSION: Treatments of combined CHM with Mifepristone or Methotrexate showed similar therapeutic effect in treating ectopic pregnancy, but the latter showed a quicker initiation and less side-effects in delaying the re-menstruation time. PMID- 12585184 TI - [Clinical and experimental study on separately decocted and mingly decocted jianweishu granule]. AB - OBJECTIVE: To observe the difference in therapeutic effect of Chinese herbal granule decocted separately and mingly. METHODS: One hundred patients of functional dyspepsia of Spleen deficiency and Liver stagnancy type were treated with Jianweishu Granule (JWSG, a self-formulated recipe by the author). Half of the patients received JWSG decocted separately, half of them received that decocted mingly. The therapeutic effects between the two groups were compared. Experimental observation on the effects of differently decocted remedies in rats was also conducted. RESULTS: The therapeutic effects obtained in the two groups were similar, with respective cure rate of 72% and 70% and the total effective rate 96% in both group. There was insignificant difference between the two groups (P > 0.05). Experimental study all showed no difference between the differently decocted remedies in inhibiting gastric acid, pepsin activity, gastric function regulation, small intestine movement improvement and pain alleviation in rats. CONCLUSION: The therapeutic effects of JWSG decocted separately or mingly are the same. This fact provides a scientific basis for clinical use of granule form of single Chinese herbs. PMID- 12585185 TI - [Effect of garlicin on adhesion molecules expression and deformability of peripheral neutrophils in patients with acute cerebral infarction]. AB - OBJECTIVE: To observe the effect of garlicin on adhension molecules CD11a and deformability of peripheral neutrophil in patients with acute cerebral infarction (ACI). METHODS: Neutrophils were separated from peripheral blood of healthy subjects and ACI patients, and incubated in 37 degrees C in vitro. The CD11a expression was detected by antibody fluorescence labeling method and the time of neutrophils passing millipore membrane were measured for calculation of the filter index. RESULTS: CD11a expression rate in healthy subjects was 34.64 +/- 25.34%, while in patients was 55.35 +/- 30.54%, difference between them was significant (P < 0.05). After garlicin treatment, it lowered to 49.16 +/- 31.68%, as compared with untreated group, P < 0.05. The neutrophil filter index in healthy group, untreated group, garlicin treated group and Nimodipine treated group was 0.87 +/- 0.46, 6.42 +/- 6.40, 3.47 +/- 3.67 and 5.03 +/- 3.72 respectively, comparison between that in the garlicin treated group and in untreated group showed significant difference (P < 0.05). CONCLUSION: Garlicin could effectively inhibit the CD11a expression in peripheral blood neutrophils and improve the deformability of the neutrophils in ACI patients. PMID- 12585186 TI - [Effect of shenmai injection on diaphragmatic fatigue in children with respiratory failure]. AB - OBJECTIVE: To study the effect of Shenmai injection (SMI) on diaphragmatic fatigue in children with respiratory failure. METHODS: Thirty-five cases of children respiratory failure with diaphragmatic fatigue were divided into two groups. The control group was treated with comprehensive therapy including anti infection, oxygen inhalation and parenteral nutrition, etc. The SMI group was treated with SMI intravenously, besides the comprehensive therapy as in the control group. Taking electrical impedance respirogram (IRG) as criterion of therapeutic effect, the effective cases after 30 min medication, time for diaphragmatic fatigue disappearance, as well as arterial blood gas analysis before and after treatment were analyzed. RESULTS: (1) In 30 min after medication, the effective cases in the SMI group (15/18) were more than that in the control group (4/17, P < 0.01); (2) Blood pH increased and PaCO2 decreased in both groups after treatment, but the decrease of PaCO2 was more significant in the SMI group (P < 0.05); (3) Time of diaphragmatic fatigue disappearance in the SMI group was shorter than that in the control group (P < 0.01). CONCLUSION: SMI is an effective drug for treatment of diaphragmatic fatigue in children with less adverse effect, and worthwhile for spreading in clinical practice. PMID- 12585187 TI - [Effect of fuzheng jianpi recipe on content of trace element and immune function in children with spleen deficiency anorexia]. AB - OBJECTIVE: To evaluate the efficacy and prospect of Fuzheng Jianpi Recipe (FZJPR) in treating children's Spleen deficiency anorexia (CSDA) by means of observing its effect on trace element content and immune function. METHODS: One hundred and thirty cases of CSDA were treated with FZJPR, one dose per day in decoction, orally taken, 30 days for one therapeutic course. Levels of T-lymphocyte subsets, IL-2R, immunoglobulin and trace elements were determined before and after treatment, and compared with those in the control group consisted of 60 healthy children. RESULTS: In the treated group, abnormal figures were shown in T lymphocyte subsets, especially in lowering of CD3 and CD4 count before treatment, and IgG content was reduced also. After treatment, CD3, CD4, CD4/CD8 ratio as well as IgG and IgA were improved significantly (P < 0.01), IL-2R percentage approached normal. Moreover, the levels of Zn and Fe changed significantly (P < 0.01). CONCLUSION: FZJPR could improve the trace elements content and immune function in CSDA children, so it is effective in treating CSDA. PMID- 12585189 TI - [Effect of tongxinluo capsule on plasma endothelin and calcitonin gene related peptide in patients with unstable angina pectoris]. AB - OBJECTIVE: To investigate the effect of Tongxinluo capsule (TXLC) on the patients with unstable angina pectoris (UAP). METHODS: Patients of UAP were divided into two groups, the treated group (n = 60) was treated with TXLC and the Danshen group (n = 30) was treated with composite Danshen tablet. The plasma endothelin (ET) and calcitonin gene related peptide (CGRP) were measured before and after treatment. Data were compared between the two groups and also compared with those measured in 20 healthy subjects for control. RESULTS: ET level was higher and CGRP level lower significantly in UAP patients than that in healthy subjects significantly (P < 0.05). After TXLC treatment, ET significantly lowered (P < 0.01) and CGRP increased (P < 0.05). But in the Danshen group after treatment, the former decreased (P < 0.05), while the latter remained unchanged (P > 0.05). The effect of TXLC was better than that of composite Danshen tablet. CONCLUSION: TXLC is an effective drug for UAP treatment, which could efficiently regulate the ET and CGRP metabolism and ameliorate the degree of myocardial ischemia damage. PMID- 12585188 TI - [Effect of Tripterygium wilfordii polyglycoside on serum IL-2 and TNF-alpha in patients with acute anterior U-veitis]. AB - OBJECTIVE: To explore the effect of Tripterygium wilfordii polyglycoside (TWP) on level of serum interleukin 2 (IL-2), and tumor necrosis factor alpha (TNF-alpha) in patients of acute anterior uveitis (AAU). METHODS: Patients of AAU were randomly divided into two groups. The treated group (n = 50) was mainly treated with TWP and the control group (n = 50) treated with bimolani. The level of IL-2 and TNF-alpha before and after treatment were determined and compared between the two groups and also compared between the treated group and normal group consisted of 50 healthy subjects. RESULTS: Levels of IL-2 and TNF-alpha in the treated group before treatment were 1.31 +/- 0.27 micrograms/L and 1.20 +/- 0.65 micrograms/L, and after treatment 1.19 +/- 0.27 micrograms/L and 0.96 +/- 0.54 microgram/L, those in the control group were 1.31 +/- 0.26 micrograms/L and 1.22 +/- 0.66 micrograms/L before treatment and 1.20 +/- 0.27 micrograms/L and 0.98 +/ 0.51 microgram/L after treatment respectively. Comparisons of the two parameters before and after treatment in both groups showed significant difference (P < 0.05 in TWP group and P < 0.01 in bimolani group). The two parameters in both treated groups were all higher than those in the normal group before treatment (P < 0.05), but showed insignificant difference after treatment (P > 0.05). CONCLUSION: Abnormal changes of IL-2 and TNF-alpha exist in AAU patients, TWP could suppress both parameters markedly therefore has a reliable effect in treatment of AAU. PMID- 12585190 TI - [Effect of dihuang mixture on T-lymphocyte subsets in pheripheral blood and cerebrospinal fluid of patients with multiple sclerosis]. PMID- 12585191 TI - [Effect of Astragalus injection in promoting IEC-6 cell differentiation through activating ornithine decarboxylase]. AB - OBJECTIVE: To explore the mechanism of Astragalus injection (AI) in repairing mucous membrane by observing its effects on the proliferation, differentiation, migration as well as on intracellular content of ornithine decarboxylase (ODC) and polyamine in rat's small intestinal crypt-like cell line (IEC-6) in vitro. METHODS: AI was added in the IEC-6 cells after they had been cultured for 24 hours. Twelve hours after adding AI, the cells were collected to test the ODC mRNA level, ODC protein, ODC activity and the intracellular content of putrescine. Twenty-four hours after adding AI, the condition of cell proliferation and differentiation was observed. Another group of IEC-6 cells were injured after being cultured for 72 hours, and AI was added, the cell migration was observed at 24 hours, 48 hours and 72 hours after medication. RESULTS: AI could inhibit IEC-6 cell proliferation, promote the cell differentiation, but with no apparent effect on injured IEC-6 cell migratin. AI of 62.5-250 micrograms/ml concentration could increase the ODC mRNA level, as compared with control, the difference was significant (P < 0.05-0.01). ODC activity and intracellular content of putrescine could be gradually increased by AI in a dose dependant manner. No effect of AI was shown on ODC protein in various dosage of AI groups. CONCLUSION: AI could promote IEC-6 cell differentiation by means of inducing the ODC activity and biosynthesis of polyamine, but without significant effect on cell migration. PMID- 12585192 TI - [Comparative study of modulating effect of kidney tonifying recipe and spleen invigorating recipe on T-lymphocyte apoptosis in corticosterone treated rats]. AB - OBJECTIVE: To compare the modulating effect of Kidney tonifying recipe (KTR) and Spleen invigorating recipe (SIR) on T-lymphocyte apoptosis (TLA) in corticosterone treated rats (CTR). METHODS: Qualitative and quantitative analysis on activation-induced TLA was conducted by transmission electron microscope and TUNEL labelled flow cytometry detection. RESULTS: TLA percentage of CTR was 45.87 +/- 7.22%, while that of normal control rats was 34.25 +/- 6.47%, the difference between them was significant (P < 0.01). TLA percentage of Youguiyin treated rats was 35.90 +/- 7.39%, and that of Bushen Yishou capsule treated rats was 36.20 +/- 9.14%, compared with that of CTR, P < 0.01 and P < 0.05 respectively. TLA percentage of the SIR treated group was 36.92 +/- 11.82%, which was different insignificantly as compared with that of CTR. CONCLUSION: TLA susceptibility was significantly enhanced in CTR. Both KTRs (Youguiyin and Bushen Yishou capsule) had down-regulating effect on TLA, while the effect of SIR was insignificant, suggesting that down-regulating activation-induced TLA may be one of the important mechanisms of KTR in improving T-lymphocyte function in CTR. PMID- 12585193 TI - [Influence of long-term use of low dose caulis Aristolochiae manshuriensis on partial nephrectomized rats]. AB - OBJECTIVE: To study the effect of long-term use of low dose Caulis Aristolochiae Manshuriensis (CAM) on the deterioration of chronic renal failure rats. METHODS: The 5/6 nephrectomized Wistar rats were taken as animal model of chronic renal failure, which were divided into 3 groups. Group A was treated with 1 g/kg CAM decoction, the dose equivalent to that defined in the pharmacopoeia. Group B was treated with 3 g/kg CAM decoction and Group C treated with equal volume of tap water. Medication was given once per day by gastrogavage in all the three groups for 8 weeks. The serum creatinine, urea nitrogen, urinary protein content and morphological changes of kidney were observed. RESULTS: After 8 weeks treatment, levels of serum creatinine, urea nitrogen, urinary protein excretion in Group B were higher than those in Group C significantly, they were 165.0 +/- 15.6 mumol/L vs 109.8 +/- 10.0 mumol/L, 27.8 +/- 3.6 mmol/L vs 18.7 +/- 2.5 mmol/L and 68.2 +/ 10.1 mg/24 hrs vs 44.6 +/- 8.5 mg/24 hrs, respectively, all P < 0.05. The pathological changes of renal mesenchyme and degree of glomerulosclerosis were also heavier in Group B. CONCLUSION: The susceptibility of chronic renal failure rats to the nephrotoxicity of CAM increases in long-term use of low dose CAM which could accelerate the deterioration of renal impairment in the model rats. PMID- 12585194 TI - [Study on induction of ginsenosides on HL-60 cell apoptosis]. AB - OBJECTIVE: To observe whether the Ginsenosides (GS) could induce HL-60 cell line apoptosis from promyelocytic leukemia. METHODS: HL-60 cells were treated with GS of various concentration to observe the effect of GS on cell morphological change, the DNA content change by flow cytometry, DNA ladder by electrophoresis, and apoptosis rate by Annexin V-FITC test of the cells. RESULTS: GS could inhibit the growth of HL-60 cells and induce cell apoptosis in a certain scope of dose and reacting time. CONCLUSION: GS could specifically induce apoptosis in HL-60 cells, which provides an experimental basis for treatment of leukemia with GS as an supplemntary agent of chemotherapy. PMID- 12585195 TI - [Extracorporeal experimental study on immuno-modulatory activity of Astragalus memhranaceus extract]. AB - OBJECTIVE: To study the effect of Astragalus membranaceus extract (AME) in regulating the immune function of human peripheral blood immune cells (PBIC) in vitro. METHODS: Effects of AME on the proliferation activity of peripheral blood mononuclear cells (PBMC) and the tumor cell phagocytosis of peripheral blood adherent monocytes (PBAM) were measured by using 3H-TdR incorporation. Effect of the tumor-killing activity of cytotoxic T-lymphocyte (CTL) was determined by using 51Cr-releasing assay. Effects on production of IgG by peripheral blood B cells (PBBC) and IL-6 by PBAM were tested by means of ELISA, and effect on production of TNF-alpha by PBAM was studied by means of biological method. Besides, the protein elements of AME were analysed by SDS-PAGE. RESULTS: AME could promote the proliferation of human PBMC, elevate the tumor cell-killing activity of CTL, strengthen the tumor cell phagocytosis and cytokines (TNF-alpha and IL-6) production of PBAM, and promote the IgG production of PBBC. AME contained multiple protein elements. CONCLUSION: AME has effect in enhancing human immuno-function and anti-tumor activity, it could be applied in clinical practice for immuno-modulation and tumor treatment. PMID- 12585196 TI - [Survey on the symptoms and risk factors in patients with hypertension]. PMID- 12585197 TI - [Clinical observation on treatment of 49 cases of cholestasia viral hepatitis by integrative medicine]. PMID- 12585198 TI - [Drug-serum chemistry from viewpoint of create new Chinese drugs]. PMID- 12585199 TI - [How to deal with the cultural factors in English translation of Chinese articles]. PMID- 12585200 TI - [The progress of studying C. spicatus]. PMID- 12585201 TI - [Exploration on the inflammatory reacting mechanism of acute respiratory distress syndrome and immuno-regulatory treatment by integrative medicine]. PMID- 12585202 TI - [Minute on the 6th session of national conference of deficiency syndrome and gerontology]. PMID- 12585203 TI - [Prions as the driving force in transmissible spongiform encephalopathies]. AB - Transmissible spongiform encephalopathies are degenerative disorders affecting the central nervous system (CNS) occurring in a variety of species. The causative agent is thought to be composed of an abnormal form of the host encoded prion protein (PrPC), termed PrPSc. The conformational change of PrPC into PrPSc can occur spontaneously, however, it can also be induced by PrPSc. Prion diseases such as bovine spongiform encephalopathy (BSE), scrapie and variant Creutzfeldt Jakob-Disease (vCJD) are most likely caused by peripheral uptake of prions. The process by which prions proceed to the CNS following peripheral uptake is referred to as neuroinvasion. Infection with prions is thought to occur in two phases: After ingestion prions first replicate in lymphatic tissue and then gain access to the CNS via peripheral nerves. Studies looking at the biochemical and clinical characteristics of BSE and vCJD demonstrated that BSE is most likely responsible for vCJD in humans. PMID- 12585204 TI - [Diagnosis of transmissible spongiform encephalopathies in animals]. AB - Histopathological examination of the central nervous system is essential for the confirmation of a TSE diagnosis. Typical lesions are spongiform changes of the grey matter, intraneuronal vacuoles in particular nuclei of the brain stem, gliosis and neuronal degeneration. The nature of the lesions is similar between species. However, the variation in the distribution and severity of the changes is striking. Even more reliable than histopathology is the detection of disease specific protease-resistant prion protein (PrPSc) using immunohistochemistry. The so-called "rapid tests" allow detection of PrPSc in unfixed tissues and are mostly used for the screening of risk populations and slaughtered animals. PMID- 12585205 TI - [Clinical signs and diagnosis of BSE]. AB - Bovine spongiform encephalopathy (BSE) in cattle is characterised by typical signs and an insidious course. There is gradual weight loss and decreased milk production, and the disease is distinguished by disturbances in behaviour, sensitivity and locomotion. The most common behavioural changes are apprehension and abnormal temperament. The most common changes in sensitivity are hypersensitivity to tactile, auditory and optical stimuli, kicking during milking and excessive licking of the muzzle. The most important disturbances in locomotion include ataxia, falling down and inability to get up. Diagnostic procedures include a thorough clinical and neurological examination followed by a more detailed examination to assess behaviour, locomotion and sensitivity to touch, sound and light stimuli. Other causes of central nervous system disease may be ruled out by a complete blood count, biochemical profile, urinalysis and examination of cerebrospinal fluid. Disturbances in behaviour, locomotion and sensitivity that occur singly, rather than in combination with each other, should be interpreted with caution. However, in cattle with disturbances in two or more of these categories, a tentative diagnosis of BSE can be made, and the veterinary authorities must be notified. PMID- 12585206 TI - [A method to identify BSE suspects in emergency slaughtered cattle during ante mortem examination]. AB - A method is presented by which a maximal number of BSE-infected cattle that had escaped the filter of clinical examination are identified during ante mortem examination. This approach might prove to be an efficient and cost-effective method for veterinary and non-veterinary meat inspectors to remove infected animals prior to slaughter. In a case-control study, the clinical signs of 224 randomly selected sick slaughtered animals were compared with the clinical signs of 26 sick slaughtered animals in which BSE infection was diagnosed using a rapid test post mortem. In addition, the clinical signs of the sick slaughtered BSE positive animals were compared with the clinical signs of a group of BSE suspects identified during the same time period and in which BSE infection could be confirmed. As a result of this study a mathematical model was developed to identify BSE suspects. This model contains a total of 7 variables (clinical signs) that proved to be of importance. These signs were aggressive behaviour, grinding of teeth, protruding eyeballs, reduced rumination, inability to stand, overexcitability, and difficulty in standing up. The presented model has a sensitivity of 61.5% (16 out of 26 BSE-positive animals slaughtered while sick were identified) and a specificity of 99.6% when compared to a rapid test conducted post mortem. PMID- 12585207 TI - [Beyond BSE: Transmissible spongiform encephalopathies in other animal species]. AB - There are several other diseases besides BSE which belong to the group of transmissible spongiform encephalopathies (TSE). Although most mammals can be experimentally infected with the agent of these diseases, generally only single representatives of the orders Artiodactyla (cloven-hoofed animals), Carnivora (carnivores) and Primates (humans and monkeys) are naturally infected in the field. An overview of the current state of knowledge on TSE in several species like exotic ruminants, deer, mink and cats is presented. Etiological, clinical, anatomic-pathological and epidemiological aspects are described. PMID- 12585208 TI - [Transmissible spongiform encephalopathies in sheep and goats]. AB - Scrapie has been known to occur in sheep and goats for over 250 years. Sheep and goats can be experimentally infected orally with BSE, however BSE infection can only be distinguished from scrapie using costly and extensive laboratory procedures. An overview of both the current state of knowledge on Transmissible Spongiform Encephalopathies (TSE) in small ruminants as well as the TSE monitoring and control strategies in place in Switzerland is presented. PMID- 12585209 TI - [Transmissible spongiform encephalopathies in humans]. AB - Transmissible spongiform encephalopathies (TSE) are dementing diseases and have been known to affect humans for over 90 years. The most common of these is the sporadic form of Creutzfeldt-Jakob disease (sCJD), followed by its familial (fCJD) and an iatrogenic (iCJD) form. 1996 a variant of CJD (vCJD) has been described in the UK, of which so far 131 cases have been observed worldwide. Specific biochemical and neuropathological signatures allow to distinguish between vCJD and sCJD and lead to the hypothesis that vCJD is due to transmission of BSE prions to humans. Although promising therapeutical approaches are being investigated, human TSE remain untreatable entities. Thus preventive measures are essential. In Switzerland the population has been exposed to BSE prions, too, but no case of vCJD as described in the UK has been observed until now. Since 2001, however, a so far unexplained increase of sCJD cases is being observed. PMID- 12585210 TI - [The eradication of BSE: a touch stone for the veterinary services]. AB - The eradication of BSE is proving to be a lengthy task. Undisputedly, it has become one of the highest priority tasks of the Veterinary Services in Europe since the disease first occurred in the mid eighties. However, the effects of the ongoing eradication effort also impair important economic agricultural sectors. Risk analysis has never before been such a critical tool in the decision-making processes for eradication of animal diseases, despite the problem that many risk analyses are being conducted in the face of incomplete knowledge about BSE. In the focus are the measures minimizing the risk for humans (such as the ban on specified risk material) and the measures implemented to prevent the spread of the disease in animals with the ban on feeding meat and bone meal to ruminants. Due to the long incubation period, implementation of insufficient measures may not be noticed for many years. Implementation of drastic measures is needed, if the eradication of the disease is to be pursued purposefully. PMID- 12585211 TI - [Geographic distribution of BSE in Switzerland]. AB - Visual evaluation of the occurrence of BAB (born after ban) cases pointed towards spatial clustering. Therefore a statistical analysis of spatial case clustering was conducted using GIS technology. In the initial analysis, all 376 cases (135 BAB, 241 BBB/born before ban) diagnosed through mid of March 2001 were investigated using the spatial scan statistic. Two clusters of BBB cases were identified in the western part of Switzerland, and two clusters of BAB cases in the eastern part. Epidemiological investigations performed within the BAB clusters showed an increased pig density in these cluster regions. Pig density is considered an indicator for the probability of contamination of cattle feed with feed containing meat-and-bone meal that is intended for other species (cross contamination). In an updated cluster analysis including all cases reported until end of June 2002 (data set B), clusters were identified in the same regions as previously. It was shown that the BAB clusters occurred in different time periods. The small scale differences in cluster size and location are discussed, with an emphasis on the implications for following epidemiological investigations. PMID- 12585212 TI - [Risk assessment as an indicator for the distribution of BSE in the world]. AB - For a long time, BSE was considered a problem of the UK exclusively. Even after the detection of BSE cases in countries outside the UK, the risk of having BSE was categorically denied by many other countries. Only after the introduction of active surveillance did several "BSE free" countries detect BSE in their country. However, before the detection of the first cases in several of these countries, a risk assessment (GBR) conducted by the Scientific Steering Committee of the EU showed that a risk could be present. There remain many countries with an unknown BSE risk. In order to minimise import risks from these countries, further risk assessment is needed to evaluate the real BSE distribution worldwide. PMID- 12585213 TI - [Twelve years experience in Switzerland: Quo vadis BSE?]. AB - When the first BSE case in Switzerland became public in 1990, it precipitated a chain of events with many diverse consequences. During this time, we continued to gain knowledge, implement new measures, and make practical recommendations in order to attempt to control the disease. This was made possible by the establishment of a huge data base containing information on all cases of BSE that had been diagnosed in Switzerland, as well as by the use of information from experiences in other countries. Analysis of these data allowed the sequence of events to be described temporally and spatially and an epidemiological evaluation to be made. These data also allow predictions to be made regarding the future of BSE in Switzerland, although these predictions must be interpreted cautiously. PMID- 12585214 TI - [Bioresorbable barrier membranes for guided bone regeneration around dental implants]. AB - The aim of this study was to evaluate the efficacy of two bioresorbable barriers especially produced for guided bone regeneration. Six beagle dogs were used in this investigation. At the beginning of the study, all mandibular premolars were extracted and after a healing period of three months three screw-type machined implants were inserted in each side of the mandible. At the buccal aspect of each implant bed a dehiscence type defect with a coronoapical and mesiodistal extension of 5 mm was created. Then, one of the following four methods for defect treatment was applied: 1) guided bone regeneration (GBR) with a bioresorbable barrier (poly-L/DL-lactide) (test group 1), 2) GBR with a bioresorbable composite barrier [poly-L/DL-lactide reinforced with tricalciumphosphate (TCP)] (test group 2), 3) GBR with an expanded polytetrafluoroethylene barrier (GTAM) (control group 1), and 4) no treatment (control group 2). The animals were sacrificed six months after implant installation. During the healing period the most common problems encountered in the test groups 1 and 2 were soft tissue dehiscences and bioresorbable membrane exposures. Retrieved tissue specimens were processed for histological and histometric evaluation. The mean amount of vertical bone regeneration at the defect areas was 1.75 mm for test group 1, 1.82 mm for the test group 2, 2.38 mm for control group 1, and 1.93 mm for control group 2. When the amount of bone regeneration in contact to the implant surface was evaluated, the regeneration result was 1.45 mm for test group 1, 1.49 mm for test group 2, 2.08 mm for control group 1, and 0.91 mm for control group 2. For both measurements, no statistically significant differences could be observed between the different groups. The present animal experiment showed that the treatment of dehiscence type defects around machined implants with the two bioresorbable barriers showed only slightly superior results compared to the treatment without barriers (control group 2). The best results--although not statistically significant different from the other treatment groups--were obtained when nonresorbable barriers were used. PMID- 12585215 TI - [Controlled clinical study of the anterior composite-adhesive system Point 4/OptiBond Solo Plus]. AB - A wide variety of presently sold composites exhibit a large diverse range of colors and yield the desired esthetic result by the "Chameleon Effect" and by the application of a thin layer technique. The present clinical trial of the ultra small-particle-hybrid composite Point 4 with its dentin adhesive OptiBond Solo Plus just reached the one-year evaluation. Of the originally applied 73 restorations of anterior teeth in the classes III to V, 62 could be re-examined after half a year and 60 after one year according to an examination scheme by Lutz et al. (1977) and Ryge (1980), modified by Bohm et al. (1991). While the color of restorations and the amazingly smooth surface only changed slightly, significant differences could be established in the criteria bond zone morphology and volume behaviour after half a year and one year compared with baseline. Results showed retention rates of 98% (2% total loss) and 97% (3% inclusive partial loss) as well as initial marginal discolorations of 20%. Thus, the moist bonding technique, applied here, appeared less suitable than the wet-bonding technique. PMID- 12585216 TI - [Minimally invasive therapy based on improved periodontal diagnosis. Case report]. PMID- 12585217 TI - [A new splint technique in dental traumatology]. AB - The treatment of displacement injuries and fractures of permanent teeth is an important emergency therapy in the dental office. Due to new sport trends and various outdoor activities, the frequency of dental trauma of children and adolescents alike is steadily rising. The standard treatment of displacement or avulsion injuires of permanent teeth is repositioning or replantation with subsequent splinting. The dental trauma splint must be flexible to allow for the physiologic movement of the repositioned or replanted teeth in order to reduce the risk for ankylosis or external root resorption. Stabilization of a repositioned tooth with a flexible splint over a short time period optimises the rehabilitation of the periodontal tissues. In the literature various splinting techniques have been described. This paper presents a new splinting technique; the Titanium Trauma Splint (TTS), which is made of pure titanium and was specifically developed for the treatment of traumatically displaced or avulsed permanent teeth. The TTS-splint has certain advantages when compared to traditional splinting techniques. These advantages are beneficial both for the dentist and patient alike. PMID- 12585218 TI - HIV/AIDS: mainstream or forgotten? PMID- 12585219 TI - Reducing HIV sexual risk among African American women who use drugs: hearing their voices. AB - A pilot study was conducted to understand women's realities, perspectives, and perceived needs regarding the interaction between substance use, types of sexual relationships, and the challenges and barriers they pose to practicing safer sex. An additional purpose was to examine this population's willingness to participate in the program and receptiveness to the components of proposed interventions. Data were collected in this descriptive, exploratory study by the use of one focus group. Twelve African American women between 20 and 55 years of age who had previous drug detoxification experience participated in a 3-hour focus group. All participants were sexually active as well as current users of illicit substances, and all but one had traded sex for money, drugs, or alcohol. The focus group interview was guided by a semistructured questionnaire. Participants were recruited by counselors in detoxification units. Each unit counselor was asked to randomly select a third of their residents, explain and read the purpose of the focus group, and get the residents' verbal consent to participate. The women discussed their sexual behaviors and drug use that increased their risk for acquiring HIV. They provided insight about the effects of different drugs on one's sexual behavior, motivation for sex, and reasons for unsafe sex and made recommendations for HIV prevention intervention. Participants agreed that safer sex and healthy living intervention would be most beneficial if placed within the context of a detoxification program. The juxtaposition of increased sexual risk behavior and the potential to acquire and transmit HIV infection suggests the urgency of intervening with African American women who are substance abusers. PMID- 12585220 TI - The interrelationship between violence, HIV/AIDS, and drug use in incarcerated women. AB - The purpose of this study was to examine how violence co-occurs with drug use and HIV/AIDS high-risk behaviors among women incarcerated for drug use. This study was a supplement to a larger, experimental study designed to reduce HIV/AIDS high risk behaviors among incarcerated female drug users. The women who participated (N = 170) reported that violence was a major part of their lives and affected many of their behaviors. Among these 170 women, 26.6% used condoms for oral sex, 46.4% for vaginal intercourse, and 65% were either sexually or physically abused. Intimate partner abuse was reported by 33.9% of the women. About 16% reported forced sexual activity, and 17.5% feared their partners. Women who refused to give oral sex to their male partners and those who insisted on condom use during oral sex prior to jail were more likely to be sexually or physically abused (chi 2 = 4.104; chi 2 = 3.886, p < .05, respectively). Although statistically significant interrelationships were not found among the three variables, significant bivariate relationships were found between intimate violence and HIV/AIDS high-risk behaviors. PMID- 12585221 TI - Context-based advocacy for HIV-positive women making reproductive decisions. AB - As the number of HIV-positive women of childbearing age continues to rise, and treatments available to manage HIV become more accessible, the issue of HIV positive women's reproductive decision making is gaining importance for nurses in AIDS care. Nurses and other health professionals care for these women as they decide whether to bear children. The decision whether to have children is complex and influenced by a number of individual and societal factors, creating an ethical tension between the interests of HIV-positive women and those of society. This article proposes a six-step, context-based advocacy process for nurses and other health professionals who interact with HIV-positive women of childbearing age. The advocacy process described is grounded in a review of literature on HIV/AIDS, factors influencing HIV-positive women's reproductive choices, and nursing advocacy. The proposed advocacy process enables nurses to support women's self-determination and decision making in a way that is informative and empowering. PMID- 12585222 TI - Buddy programs for people infected with HIV. AB - The purposes of this correlational study were to describe and compare clients' and volunteers' perceptions of a buddy program for people infected with HIV and to identify relationships between social support and clients' quality of life. Clients' social support was assessed with the Interpersonal Support Evaluation List(ISEL), and their quality of life was assessed with the Medical Outcomes Study-HIV (MOS-HIV) Scale. Clients' and volunteers' perceptions of satisfaction and assistance with activities were assessed with researcher-developed instruments. A convenience sample of 46 client-volunteer dyads was recruited from five AIDS service organizations. Clients perceived adequate levels of social support, moderate amounts of assistance, high levels of satisfaction with client volunteer relationships, and moderate to low quality of life. A positive relationship was found between clients' and volunteers' perceptions of satisfaction. Relationships were found between ISEL subscales and the Health Transition and Mental Health subscales of the MOS-HIV and the MOS-HIV total scale scores. The findings of the study provide support for the continuation of buddy programs. PMID- 12585223 TI - The physiologic health care needs of HIV-infected black men on admission to an AIDS-dedicated nursing home. AB - The purpose of this study is to describe the physiologic health care needs of HIV infected Black men on admission to an AIDS-dedicated nursing home. Participants were 68 Black men aged 26 to 60 years who were admitted to an AIDS-dedicated nursing home in the southern New England area between 1995 and 1999. The participants were very ill and weak on their admission to the nursing home, with most having diagnoses of AIDS (n = 65), an average Karnofsky Performance Scale score of 44 (SD = 14.90), and some degree of mental impairment. The late-stage of disease of the participants was reflected in their multiple symptomatology and functional impairment in activities of daily living. With patients living longer in the chronic disease stages of HIV disease, the results of this study provide support for the further investigation of the most effective long-term care settings for Black men with HIV/AIDS. The results also have implications for the multiple clinical roles nursing can assume within HIV/AIDS long-term care settings. PMID- 12585224 TI - Clinical implications of therapeutic exercise in HIV/AIDS. PMID- 12585225 TI - HIV prevention: a review of interventions. PMID- 12585226 TI - Transmission of HIV infection among older adults: a population at risk. PMID- 12585227 TI - Changing hostile attitudes toward homosexuals in a region facing one of the highest incidences of HIV/AIDS. PMID- 12585228 TI - Protease inhibitor shown to hold HIV at undetectable levels. PMID- 12585229 TI - Electron microscopy of umbilical cord endothelial cells in preeclampsia. AB - We have studied by means of transmission electron microscopy the umbilical cord endothelial cells in 11 preeclamptic women. Endothelial cells exhibited oval, round, elongated, flattened, triangular or polygonal shapes. The nuclei displayed shallow and deep invaginations of nuclear envelope. The endoplasmic reticulum appeared highly dilated and vacuolated. The swollen mitochondria showed cristae fragmentation. Areas of focal necrosis were appreciated throughout the cytoplasm. A marked enlargement of subendothelial space and the presence of an electron dense granular material were also found. The findings reveal activation and injury of endothelial cells and disruption of endothelial cell layer. PMID- 12585230 TI - Morphology of the compound eyes of Nebalia herbstii Leach, 1814 (Leptostraca, Nebaliidae). AB - The Nebalia compound eyes are scotopic apposition eyes that provide a roundabout view. Each ommatidium is equipped with a large, layered retinula, a flat cornea, a eucone crystalline cone made of four cone cells, and eight retinula cells arranged in three cell pairs that exhibit mirror symmetry (R1/R6, R2/R5 and R3/R4) as well as two single cells (R7 and R8). Cone cell threads run between retinula cells R1 and R2, R3 and R4, R5 and R6, R8 and R1. A plane of symmetry divides the compound eyes into anterior and posterior parts containing ommatidia arranged in mirror symmetry and having different R-cell chirality. The findings are discussed with respect to the structural features of other crustacean eyes and basic features of the arthropod compound eye. PMID- 12585231 TI - Pharmaco-electroencephalography and pharmacokinetic-pharmacodynamic modeling in drug development: focus on preclinical steps. PMID- 12585232 TI - In-house diabetes program delivers on a 'shoestring' budget. AB - Companion HealthCare, a Columbia, SC-based subsidiary of BlueCross BlueShield of SC, has been able to achieve impressive results by building its own in-house DM programs. Funded "on a shoestring,"these programs have proven far less expensive to implement than vendored models while still producing impressive clinical returns. PMID- 12585233 TI - Evaluation of antiherpesvirus-1 and genotoxic activities of Helichrysum italicum extract. AB - The antiherpes virus-1 and genotoxic activities of diethyl ether extract from flowering tops of Helichrysum italicum (Compositae) were investigated. The extract showed significant antiviral activity at concentrations ranging from 400 to 100 microg/ml. This activity was not due to cytotoxic effect of the extract since Vero cells exhibited altered morphology or growth characteristics indicative of cytotoxic effects at higher concentration (800 microg/ml). Moreover H. italicum extract showed no DNA-damaging activity at concentrations up to 2000 microg/disk. PMID- 12585234 TI - Polysaccharides of higher plants as factors of secondary metabolism regulation. PMID- 12585235 TI - Spermatogenesis in mice carrying Dominant Spotting Yurlovo, a c-kit gene mutation. PMID- 12585236 TI - The first data on the intestine nervous system organization in priapulids. PMID- 12585237 TI - Evaluation of the correlation and regression relationships between the concentrations of alkaloids and chemical elements in individual greater celandine (Chelidonium majus L.) plants. PMID- 12585238 TI - [Potentialities of spiral computed tomography in the diagnosis and treatment of traumatic injuries of the midface]. AB - The paper summarizes the results of examination of 169 patients referred to the surgeon in the period from 1 till 47 days after medial facial (MF) trauma by using spiral computed tomography (SCT) and the method of laser stereolitography (LSLG). The actual aspects of diagnosis and treatment are marked; the methods for the analysis of injuries of soft tissue structures and adjacent areas while orbit trauma are developed together with the methods for determination of degree of enophthalmos. Also discussed are the possibilities of SCT and LSLG in visualization of MF injuries and planning patient treatment. PMID- 12585239 TI - Spontaneous cerebrospinal fluid otorrhoea: case report and literature review. AB - Cerebrospinal fluid otorrhoea may occur through the temporal bone both in children and adults. In children it is generally associated with labyrinthine malformations and usually presents with hearing loss in a child with recurrent meningitis. In adults it is sequel to direct head injury, otologic or neurotologic surgery or infection. More rarely this pathology is described as being "spontaneous", occurring without any history of trauma, surgery or infection. Spontaneous cerebrospinal fluid otorrhoea in adults may present with dull symptoms such as a blocked ear or short term conductive hearing-loss. The anatomic site of this fistula is the tegmen tympani which may have a microscopic or macroscopic bone deficiency or sometimes even a "silent" meningoencephalic herniation. The authors describe a case of spontaneous cerebrospinal fluid otorrhoea occurring in an adult patient with bilateral absence of the tegmen tympani and review the literature regarding this specific. They suggest that its actual occurrence may be underestimated. Special attention should be given to adult patients with recurrent or persistent middle ear effusion. Any suspicion should be followed by meticulous imaging and surgical exploration since this may be a lifethreatnening situation. PMID- 12585240 TI - Leptospirosis presenting with encephalitis-induced coma. PMID- 12585241 TI - [The training of public school cafeteria staff: an analysis of the instructional material developed by Instituto de Nutricao Annes Dias - Rio de Janeiro (1956 1994)]. AB - The present study aims at analyzing educational approaches in the training of public school cafeteria staff members from 1956 to 1994 through the study 17 instructions brochures developed by Instituto de Nutricao Annes Dias in the municipality of Rio de Janeiro. The analysis of the documents has considered two periods: the first one, from 1956 to 1971, is characterized by the foundation of the institute and its first training activities; the second period goes from 1972 to 1994, which is characterized by the consolidation of the institute's organizational structure and the presence of educational concepts in its training activities. Training focused on technical aspects and recurrently alluded to hygiene and organization. Discussions on the training of school cafeteria staff members should define what kind of workers is expected before defining what skills and abilities they are expected to develop. PMID- 12585243 TI - Rachel's plea and donor insemination. PMID- 12585244 TI - [Contra-indications to taking estrogen]. PMID- 12585245 TI - If this is the information age, why doesn't anyone know anything? PMID- 12585246 TI - Optional Caesarean: what do some Canadian physicians say? PMID- 12585247 TI - A personal recollection of Virginia Apgar. PMID- 12585248 TI - Ambulant? Continent? The missing link. PMID- 12585249 TI - Re: Leader, A. Rachel's plea and donor insemination. JOGC 2002;24(2:125-6). PMID- 12585251 TI - Neurophysiological analysis of leg movements in infants with spina bifida aperta in the early postnatal period. PMID- 12585250 TI - Caudal regression syndrome: an online multicentre survey. Urological long-term results. PMID- 12585252 TI - Parent and school perceptions of language abilities in children with spina bifida and shunted hydrocephalus. PMID- 12585253 TI - Application of the Token Test with myelomeningocele and shunted hydrocephalus. PMID- 12585254 TI - Deep venous thrombosis in individuals with spina bifida. PMID- 12585255 TI - How to improve primary prevention of neural tube defects in Flanders: perspectives of the target population, care providers and stakeholders. PMID- 12585256 TI - The incidence and pattern of neural tube defects in Dar es Salaam, Tanzania. PMID- 12585257 TI - Outcome of hydrocephalus and spina bifida surgery in a referral hospital without neurosurgical services in Tanzania. PMID- 12585258 TI - The efficacy of bowel management in the adult with spina bifida. PMID- 12585259 TI - Pitfalls in perinatal shunt surgery: a personal perspective. PMID- 12585260 TI - Subcommissural organ dysfunction in H-Tx rats with early-onset hydrocephalus. PMID- 12585261 TI - Enhanced expression of the NA-K-2 Cl cotransporter at different regions of the blood-CSF barrier in the perinatal H-Tx rat. PMID- 12585262 TI - The "failures" of spina bifida transdisciplinary care. PMID- 12585263 TI - Headaches in children with shunted hydrocephalus: an investigation of psychological factors. PMID- 12585264 TI - Nutrition for cancer patients -- are there specific recommendations. PMID- 12585266 TI - Do we need experimental oncology? PMID- 12585267 TI - AHRQ researchers examine the role of informatics in responding to bioterrorism, mass disasters, and war. PMID- 12585268 TI - Federal, State and local law enforcers tackle deceptive spam and internet scams. PMID- 12585270 TI - Abstracts of the XV Annual Meeting of the Latin-American Society of Pediatric Endocrinology. 27-31 October 2002, Punta del Este, Uruguay. PMID- 12585269 TI - Abstracts of the Australian Conference of Science and Medicine in Sport. 12-16 October 2002, Melbourne, Australia. PMID- 12585271 TI - [Spontaneous generation and the hygienic concern about germ dissemination]. AB - This paper analyzes the answer given by Pasteur to those who defended the spontaneous generation theory and evaluates the impact these arguments had on the history of public hygiene. Because of that, the article emphasizes transmission of germs, rather than the origin of life, which has been quite analyzed. It also discusses the criticism Pasteur made to naturalists' arguments and tries to show that it is possible to say there is a certain continuity between the air-related concerns on the part of classic hygienists, most of which infectionists, and Pasteur's concern with germs spread in the air. PMID- 12585272 TI - [Berlin-Brandenburg Ophthalmologist Society Meeting. 30 November-1 December 2002. Abstracts]. PMID- 12585273 TI - [Saxony Ophthalmologist Society Meeting. Dresden, Germany, 16-17 August 2002. Abstracts]. PMID- 12585275 TI - Leading the Way: The Gillette Nursing Summit on Integrated Health and Healing. May 30-31, 2002. St. Paul, Minnesota, USA. PMID- 12585274 TI - [Tuberculosis and milk: elements of a polemic]. AB - Can human beings get tuberculosis from the milk taken from cows infected with tuberculosis? Are the diagnosis and control of cattle tuberculosis effective for the protection of human populations? Questions such as these were the main concern of doctors from Sao Paulo at the beginning of the twentieth century. The present study tried to recover elements from the polemic that took place in Sao Paulo, in order to encourage the coming up of complementary facts about such matter. The article presents some indicators of confronting positions: on one hand, the goal of intensying the control of tuberculosis among the cattle; on the other hand, the attempt not to bring up any loss to national cattle raisers through the demand of additional precaution measures, which would affect the production and distribution of their products. Medical publications kept records of the interventions, which have allowed the author to identify the first position as the stronger one, which has brought up relevant consequences for the sanitary food control. PMID- 12585276 TI - Abstracts of the XIIth International Vascular Biology Meeting. 12-16 May 2002. Karuizawa, Japan. PMID- 12585277 TI - [Images of Brazil: a Rockefeller Archive Center collection]. AB - This article describes and comments on the collection of photographs of Brazil that belongs to the Rockefeller Foundation archives (Rockefeller Archive Center) in the United States. It also contains information about the origin of such documents and the way they were collected, as well as analyzes the context in which these visual documents were produced. They record the relationship between Rockefeller Foundation and Brazilian institutions engaged in the combat against yellow fever, malaria and ancyclostomaisis from 1920 to 1940. PMID- 12585278 TI - Definition of Nidana Pancaka (first chapter of Madhavanidana). AB - This is in continuation of the previous article published in this Bulletin of the Indian Institute of the History of Medicine, Volume XXVII, No. 2, July 1997. P. Nos 163 to 171, under the title of "Definition of Nidana Panchaka (First Chapter of Madhava Nidana"). It deals with Nadana, Purvarupa, Rupa, Upashaya and Samprapti as described in the first Chapter of Madhavanidana. PMID- 12585280 TI - An unusual case of oesophageal metastasis from gastric adenocarcinoma. PMID- 12585281 TI - 27th ESMO Congress, Nice, 18-22 October 2002. Iressa disappoints in NSCLC. PMID- 12585283 TI - Dietary problems in highly developed countries. PMID- 12585282 TI - Islamic medical ethics with special reference to Moalejat-e-Buqratiya. AB - The writer physicians of Islamic medicine have given top priority to the Islamic Medical Ethics in their valuable writings and left the ineffaceable footsteps in the field. Such as, Abul Hasan Ahmed bin Mohammad At-Tabari has devoted the 35th chapter of Maqala l of his precious book entitled 'Moalejat-e-Buqratiya' to medical ethics. The entire chapter reflects the Islamic approach of the scholar. In the following pages, an attempt has been made to highlight some salient features of Islamic Medical Ethics in ancient literature of Islamic Medical with special reference to Moalejat-e-Buqratiya. PMID- 12585284 TI - Ask the right questions! PMID- 12585285 TI - Proceedings of the Middle Eastern Summer Institute on Drug Use meetings: 1997 1999. PMID- 12585286 TI - Doctors in literature. AB - Both, medicine and creative literature map out the mind of mankind and expose the beauty of nature and the biological world. Literature gives insight into the inner dynamics of the human mind. Doctors of medicine know the cell-the building block of the body and mind. They can easily penetrate into the innermost sanctum of the human mind. They see things as they really are not as they themselves are. Creative writers are keen observers of everything around them. Keen observation very often tells or give clues to scientific truth. World literature offers a galaxy of those creative minds. It is a brief account of the contribution of doctors in literature over the centuries. It is, by no means, comprehensive. It is an overview. PMID- 12585287 TI - Social basis of deviant sexual behaviour: a historical perspective. AB - Sexual perversions are often a product of civilized life. Most societies permit some degree of deviant sexual behaviour as a relief from stress of routine life. Criteria for labelling such behaviour have changed over a period of time. There have been instances of normalizing deviant sexual behaviour by formation of pressure groups by the deviants e.g. homosexual clubs, particularly in Western countries. Attitude of church on masturbation & homosexuality has fluctuated from forbidding these activities to accepting these as harmless acts. Extra marital sex, premarital sex, homosexuality, fellatio, masturbation have been reported from almost all societies. Swinging and mate-swapping is more prevalent in the West. Social factors associated with deviant sexual behaviour are discussed. Incest lobbies have come up in U. S. Prostitution has also been there in all societies since antiquity. Earlier, prostitutes enjoyed a relatively higher social status. Their degradation started with the dawn of Christianity. In 1960s there was sexual revolution in U. S. with emphasis on free sex. There is evidence of slowing down of sexual revolution with the advent of AIDS. Safe-sex and fidelity are now being emphasized. PMID- 12585288 TI - Proceedings of the 19th International Papillomavirus Conference. Florianopolis, Brazil, 1-7 September 2001. PMID- 12585289 TI - European alchemy: some traditional beliefs. AB - Three important doctrines of European alchemy are discussed: the Emerald Table of Hermes, the idea of transmutation, and the Elixir of Life. The analysis of these problems is focused on the 16th century, the epoch of the high flourish of alchemy in Renaissance Europe. As typical examples two works are chosen: the treatises of Jean-Pierre Fabre (1588-1658) and Alexander von Suchten (? 1520 - ? 1590). The arguments of these authors illustrate the ways how alchemists tried to defend their position in face of repeated failures. Just the 16th century stood in the sign of dramatic development of crafts, but, simultaneously, of growing interest in alchemy. PMID- 12585291 TI - Bibliography. Current world literature. Upper airway disease. PMID- 12585290 TI - Proceedings of the 3rd International Conference on Oxygen/Nitrogen Radicals: Cell Injury and Disease. PMID- 12585292 TI - Gleanings from the travelogue of the traveller Nicholas Seen. AB - The traveller Nicholas Senn, visited India in the early part of this century, to make detailed enquiries about the development of medical science, general education and to obtain proficiency in the professions and arts. According to him revolutionary changes were taking place day to day in research and discoveries in the fields of science and also in the practice of medicine and surgery at the time. Scientific work was no longer confined to a favoured institutions, it may be seen in all parts of the civilized globe. A personal acquaintance with men who have earned a well-merited reputation in the advancement of medical profession is a source of great gratification and excites a new interest in the work he has accomplished. PMID- 12585293 TI - Bibliography. Current world literature. Mechanisms of allergy and adult asthma. PMID- 12585294 TI - Yugi's pramegam and diebetes mellitus: an analogue. AB - Yugimahamuni was one among the greatest siddhars and has matchless contribution towards enrichment of Siddha system. In his Vaidya Chinthamani-800, he has written clearly about 4448 names of the diseases, its signs, symptoms and prognosis. Under the heading - "Meganoikal," he has elaborated twenty varieties of the urinary disorders based on the physical characters of urine. But the general signs and symptoms of 'Meganoikal' described by Yugi clearly indicate that many of these characteristics are of diabetic in nature. Under the sub heading 'avasthaikal', Yugi accurately described certain sufferings experienced by the patients of 'Meganoikal'. It is quite interesting to note that those ten types of sufferings he has listed out, starting from obesity ending in Tuberculosis are comparable with the acute and long term complications of diabetics, and hence this analogue. PMID- 12585296 TI - Bibliography. Current world literature. Genetics and epidemiology. PMID- 12585295 TI - Selected articles on medical history from Hindi journals (1990-1993). PMID- 12585297 TI - Bibliography. Current world literature. Skin and eye. PMID- 12585298 TI - The impact of information technology on emergency health care outcomes. AB - We analyze the productivity of information technology in emergency response systems. "Enhanced 911" (E911) is information technology that links caller identification to a location database and so speeds up emergency response. We assess the impact of E911 on health outcomes using Pennsylvania ambulance and hospital records between 1994 and 1996, a period of substantial adoption. We find that as a result of E911 adoption, patient health measured at the time of ambulance arrival improves, suggesting that E911 speeds up emergency response. Further analysis using hospital discharge data shows that E911 reduces mortality and hospital costs. PMID- 12585299 TI - The geographical data of the Kasyapa Samhita. AB - One of the parameters in ascertaining the date of the Kashyapa Samhita is the data of geography referred to there in. Geography abounds various spheres like country, province, city, river, mountain and sea. The whole geographical material is mentioned in the chapter "Bhojana Kalpa" of the Kalpasthana, 22nd and 25th chapters of the Khilasthana. PMID- 12585300 TI - Hospital market structure and the behavior of not-for-profit hospitals. AB - I exploit a change in hospital financial incentives to examine whether the behavior of private not-for-profit hospitals is systematically related to the share of nearby hospitals organized as for-profit firms. My findings demonstrate that not-for-profit hospitals in for-profit intensive areas are significantly more responsive to the change than their counterparts in areas served by few for profit providers. Differences in financial constraints and other observable factors correlated with for-profit hospital penetration do not explain the heterogeneous response. The findings suggest that not-for-profit hospitals mimic the behavior of private for-profit providers when they actively compete with them. PMID- 12585301 TI - Concerns about mouse housing system. PMID- 12585302 TI - Does competition under Medicare Prospective Payment selectively reduce expenditures on high-cost patients? AB - Competition and prospective payment have been widely used to control health care costs but may together provide incentives to selectively reduce expenditures on high-cost relative to low-cost patients. We use patient discharge and hospital financial data from California to examine the effects of competition on costs for high- and low-cost admissions in the 12 largest Diagnosis-Related Groups before and after the Medicare Prospective Payment System (PPS). We find that competition increased costs before PPS, but that this effect decreased afterward, especially inpatients with the highest costs. We conclude that competition and PPS selectively reduced spending among the most expensive patients and that careful assessment of these patients' outcomes is important. PMID- 12585303 TI - Do pharmaceutical prices respond to potential patient out-of-pocket expenses? AB - Despite the importance of patient insurance in the market for prescription pharmaceuticals, little is known about the impact of patient reimbursement on the pricing behavior of pharmaceutical firms. I examine the link between potential patient out-of-pocket expenses and pharmaceutical pricing using a unique policy experiment from Germany. Starting in 1989, a maximum reimbursement for a given medicine replaced a flat prescription fee. This change in reimbursement exposes the patient to the price of a prescribed product. Using a product-level panel data set covering several therapeutic categories before and after the policy change, I find that producers significantly decrease prices after the change in potential patient out-of-pocket expenses. Price declines are most pronounced for brand-name products. Moreover, branded products that face more generic competitors reduce prices more. PMID- 12585304 TI - The effects of hospital ownership on medical productivity. AB - To develop new evidence on how hospital ownership and other aspects of hospital market composition affect health care productivity, we analyze longitudinal data on the medical expenditures and health outcomes of the vast majority of nonrural elderly Medicare beneficiaries hospitalized for new heart attacks over the period 1985-1996. We find that the effects of ownership status are quantitatively important. Areas with a presence of for-profit hospitals have approximately 2.4% lower levels of hospital expenditures, but virtually the same patient health outcomes. We conclude that for-profit hospitals have important spillover benefits for medical productivity. PMID- 12585305 TI - Are for-profit hospital conversions harmful to patients and to Medicare? AB - We examine how changes in hospital ownership to and from for-profit status affect quality and Medicare payments per hospital stay. We hypothesize that hospitals converting to for-profit ownership boost post acquisition profitability by reducing dimensions of quality not readily observed by patients and by raising prices. We find that 1-2 years after conversion to for-profit status, mortality of patients, which is difficult for outsiders to monitor, increases while hospital profitability rises markedly and staffing decreases. Thereafter, the decline in quality is much lower. A similar decline in quality is not observed after hospitals switch from for-profit to government or private nonprofit status. PMID- 12585307 TI - Aromatic drugs in Unani medicine with special reference to Kitabul-Mia-Lil Masihi. AB - This is an established fact since time immemorial that, the aroma plays a vital role in the human beings and even in animals. The aromatic plants and aromatic chemicals contained in them has also significance in our day to day living. Process of distillation as adopted and described by Unani physicians confirms the claim that, they were aware of the importance of aromatic drugs and perfumes. There is a vast literature scattered in existing Unani medical books, which shows their intelligential towards the knowledge of herbal drugs including aromatic plants. 'Kitabul-Mia-Lil-Masihi is a book on Unani medicine. Its 17th Chapter consists of aromatic drugs exclusively. The drugs have been classified and presented here under different headings. PMID- 12585306 TI - Managed care, technology adoption, and health care: the adoption of neonatal intensive care. AB - Managed care may influence technology diffusion in health care. This article empirically examines the relationship between HMO market share and the diffusion of neonatal intensive care units. Higher HMO market share is associated with slower adoption of mid-level units, but not with adoption of the most advanced high-level units. Opposite the common supposition that slowing technology growth will harm patients, results suggest that health outcomes for seriously ill newborns are better in higher-level units and that reduced availability of mid level units may increase their chance of receiving care in a high-level center, so that slower mid-level growth could have benefitted patients. PMID- 12585308 TI - Underweight the main cause of ill-health; five causes account for 28% of all DALYs. PMID- 12585309 TI - Health in Goa in the 17th century: the reports of some French travellers. AB - The writings of the French travellers who visited Goa in the 17th century attest to the importance of Ayurvedic medicine, although the term Ayurvedic was never used at the time. The diseases affecting Goa were not different from those at every European trading post, and the remedies used were identical. However, at the end of the century, especially after 1685, Goa's population declined sharply, partly due to the precarious living conditions there, and also as a result of competition from the Dutch and English trading companies. PMID- 12585310 TI - Extra helping of Mayo. PMID- 12585311 TI - Ethnobiology in Mizoram state: folklore medico-zoology. AB - Studies in ethnobotany and ethnozoology under the umbrella of Ethnobiology seem imbalanced in the sense that enormous publications have accumulated in case of the former but only little information has been disseminated in case of the latter. While 7500 wild plant species are known to be used by tribals in medicine, only 76 species of animals have been shown as medicinal resources (Anonymous, 1994). The present paper is the first-hand information of folklore medicine and animals in Mizoram. The animals enumerated comprise of 25 vertebrates and 31 invertebrates and are used for treatment of over 40 kinds of diseases or ailments, including jaundice, tuberculosis, hepatitis, cancer, asthma and veterinary disease. The author, however, does not recommend destruction of wild animals, be it for food or medicine. PMID- 12585312 TI - A time for change. PMID- 12585313 TI - Ayurvedic literature in Urdu (Part-II). AB - This article is a second part of previous work published with the same title in this Bulletin Vol. 28, 1998, P. nos. 151-158. It contains brief notes on six rare Ayurvedic books in Urdu. It provides two types of information, i.e. (i) Introduction to the books studied and (2) Literary history of Ayurveda of that period in which Urdu language had a prominent role in propagation of Arts and Science. It gives an idea for seeking information on these lines regarding the Ayurvedic works in Arabic and Persian also. PMID- 12585314 TI - Tax tips for transferring the family business. PMID- 12585315 TI - Options for prenatal care under Medicaid and SCHIP. PMID- 12585316 TI - Protecting the privacy of newborns. PMID- 12585317 TI - Importance of duplex scanning ultrasonography evaluation of the proximal internal thoracic artery as a coronary artery bypass graft. PMID- 12585318 TI - Contribution of the polymer standards' polydispersity to the observed band broadening in size-exclusion chromatography. AB - The contribution of the polydispersity of polymer standards to the observed band broadening in size-exclusion chromatography was evaluated. Initially, theoretical predictions based on an equation by Knox et al. were found to overestimate this contribution, greatly due to the fact that the polydispersity values specified by the manufacturers are upper limits and therefore too high to be applied in this context. An improved estimate of the polydispersity values was obtained from the size-exclusion chromatography results and these new values were used to reassess the polydispersity contribution to band broadening. For two of three columns tested the best molar-mass-distribution parameters, i.e. those the least affected by extra-column and intra-column band broadening effects, can be obtained for polymers with a molar mass in the effective range of the given column and at rather low mobile-phase flow rates. At those conditions, for low-molar-mass polymers, the estimated polydispersity index values approach the theoretical ones derived from a Poisson distribution. PMID- 12585319 TI - Capillary-channeled polymer fibers as stationary phases in liquid chromatography separations. AB - A method utilizing capillary-channeled polymer (C-CP) fibers as stationary phases in high-performance liquid chromatographic separations has been investigated. Polymeric fibers of differing backbones (polypropylene and polyester) having nominal diameters of approximately 50 and approximately 35 microm and a channeled structure on their periphery were packed into stainless steel tubing (305 x 4.6 mm I.D.) for use in reversed-phase separations of various mixtures. The fibers have eight channels running continuously along the axis which exhibit very high surface activity. As such, solvent transport is affected through the channels through wicking action. Bundles of 1000-3000 fibers are loaded co-linearly into the tubing, providing flow channels extending the entire length of the columns. As a result, backing pressures are significantly lowered (approximately 50% reduction) in comparison to packed-sphere columns. In addition, the capital costs of the fiber material (< US$0.25 per column) are very attractive. Flow-rates of up to 5 ml/min can be used to achieve near baseline separation of related compounds in reasonable run times, indicating very fast mobile phase mass transfer (C-terms). The polymer stationary phases demonstrate high selectivity for a wide variety of analytes with gradient elution employed successfully in many instances. Specifically, separations of three polyaromatic hydrocarbons (benzo[a]pyrene, chrysene, pyrene), mixtures of both organic and inorganic lead compounds [chlorotriethyllead, chlorotriphenyllead, lead nitrate, lead(II) phthalocyanine], and a lipid standard of triglycerides were accomplished on the polymeric stationary phases. Other species of biological interest, including groups of aliphatic and aromatic amino acids have also been effectively separated. The reversed-phase nature of the fiber surfaces is supported through atomic force microscopy measurements using hydrophilic and hydrophobic functionalized polystyrene beads as the probe tips. Separations of the various analytes demonstrate the feasibility of utilizing C-CP fibers as stationary phases in reversed-phase LC. It is envisioned that columns of this nature would be particularly useful in prep-scale separations as well as for immobilization matrices for organic constituents in aqueous environments. PMID- 12585320 TI - Comparison of the acidity of residual silanol groups in several liquid chromatography columns. AB - The silanol acidity of Waters Resolve C18, Waters Resolve silica, Waters Symmetry C18, Waters Symmetry silica, Waters XTerra MS C18 and underivatized XTerra columns has been measured from the retention of LiNO3 with a methanol/water (60:40) mobile phase buffered to different pH values. The Li+ cation is retained by cationic exchange with the background cation of the mobile phase (Na+) through the ionized silanols. The number of active silanols increases in the order: XTerra MS C18 << Symmetry C18 < underivatized XTerra << Resolve C18 < Resolve silica approximately equal to Symmetry silica. XTerra MS C18 does not present any residual silanol acidity up to s(s)pH 10.0 (pH in 60% methanol) as measured by LiNO3. The underivatized XTerra packing and Symmetry C18 present active silanols only at s(s)pH values higher than 7.0. For the other three columns, two different types of silanols with different acidity (s(s)pKa values about 3.5-4.6 and 6.2 6.8, respectively) have been observed. Symmetry C18 shows evidence of the presence of active basic sites that retain NO3- by anionic exchange. PMID- 12585321 TI - Study of tryptophan enantiomer binding to a teicoplanin-based stationary phase using the perturbation technique. Investigation of the role of sodium perchlorate in solute retention and enantioselectivity. AB - The retention of D,L-tryptophan enantiomers on an immobilized teicoplanin column was investigated in relation to the mobile phase sodium perchlorate concentration using the perturbation method to determine the solute distribution isotherms. From the experimental data, it appeared that the bi-Langmuir model was able to describe D- and L-enantiomer retention on the immobilized selector over the salt concentration range. An increase in the apparent enantioselectivity with an increase in sodium perchlorate concentration was observed. The chiral recognition enhancement was governed by (i) an increase in the difference of the adsorption constants for binding to the high-affinity site (aglycone pocket) between the two enantiomers and (ii) enhancement of the number of aglycone chiral regions interacting with D-tryptophan. It is suggested that an ion-pair formation mechanism between perchlorate and solute and/or selector is responsible for this behavior. In addition, this work shows that additional secondary sites on the teicoplanin surface are involved in the apparent enantioselectivity at low sodium perchlorate concentrations. PMID- 12585322 TI - Sequential fractionation procedure for the identification of potentially cytochrome P4501A-inducing compounds. AB - A multistep fractionation procedure for the separation of nonpolar aromatic compounds with respect to cytochrome P4501A induction is presented. Normal-phase HPLC on nitrophenylpropyl silica and cyanopropyl silica was tested for group specific separation as a first fractionation step. Subsequent individual compound specific PAH fractionation was done by means of reversed-phase HPLC. Electron donor-acceptor HPLC and size-exclusion chromatography were applied to separate PAHs, PCBs, PCNs and PCDD/Fs according to their number of aromatic carbon atoms, their hydrophobicity, their degree of chlorination, their planarity and their molecular size. The method was validated for complex environmental mixtures on the basis of two sediment extracts. PMID- 12585323 TI - Determination of the distribution of molecular masses of sodium hyaluronate by high-performance anion-exchange chromatography. AB - Sodium hyaluronate, or hyaluronic acid (HA), is a glucosaminoglucan used in pharmaceuticals in ophthalmic surgery and for treatment of rheumatoid arthritis. The average molecular mass of the HA polymer used in these products is often in the range 1 x 10(6)-5 x 10(6). Size-exclusion chromatography has been used for analysis of molecular masses, including the distribution, up to about 3 x 10(6). In this work, an anionic exchange chromatography method is presented by which the peak molecular mass and an estimation of the distribution of the molecular masses of HA is possible in the range 0.1 x 10(6)-5 x 10(6). PMID- 12585324 TI - Simultaneous analysis of caffeic acid derivatives and alkamides in roots and extracts of Echinacea purpurea by high-performance liquid chromatography photodiode array detection-electrospray mass spectrometry. AB - High-performance liquid chromatography (HPLC) coupled with UV photodiode-array detection and electrospray ionization mass spectrometry was developed for the simultaneous analysis of caffeic acid derivatives and alkamides in the roots and extracts of Echinacea purpurea. Caffeic acid derivatives and alkamides produced very abundant peaks in the total ion current chromatogram during negative and positive cone voltage switching. Cichoric acid and the isomer pair, dodeca 2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide, were used as a standard for quantification of caffeic acid derivatives and alkamides in E. purpurea. This novel method surpasses previously published ones in product quality control and providing the HPLC chromatographic fingerprints of biological active components in E. purpurea. PMID- 12585325 TI - Hydrophobicity gradient columns for the separation of trypsin inhibitor by hydrophobic interaction chromatography at low salt concentration. AB - We investigated hydrophobicity gradient columns composed of two columns packed with supports of different hydrophobicities in order to save time in protein separation by hydrophobic interaction chromatography at low salt concentration using a crude sample of trypsin inhibitor as a model sample. One of the two hydrophobicity gradient columns was packed with a support whose hydrophobicity was critically controlled for target protein (trypsin inhibitor) and the other was packed with a support which was less hydrophobic than the critically controlled hydrophobicity support. It was found that the hydrophobicity gradient columns are useful to separate samples containing impurities of a wide range of hydrophobicities within a reasonable time. PMID- 12585326 TI - Comparison of the differential isosteric adsorption enthalpies and entropies calculated from chromatographic data. AB - The differential isosteric enthalpies, -deltaH(ads), and entropies, -deltaS(ads), of adsorption were calculated taking the retention times of the peak maxima and the centres of gravity of peaks into account and compared with the results obtained from the adsorption second virial coefficients. A mathematical link between the -deltaH(ads) and -deltaS(ads) magnitudes and experimental data was derived through the Antoine-type equation which enables the -deltaH(ads) and deltaS(ads) magnitudes to be found from adsorption second virial coefficients, B2S, calculated on the basis of chromatographically determined adsorption isotherm data. The virial coefficients were calculated employing the values of the Toth and Unilan equation parameters. There are no significant differences to be found between the isosteric enthalpies obtained, whereas the values of the adsorption entropies were the highest for the centre of peak gravity data. PMID- 12585327 TI - Determination of stale-flavor carbonyl compounds in beer by stir bar sorptive extraction with in-situ derivatization and thermal desorption-gas chromatography mass spectrometry. AB - A method for the determination of stale-flavor carbonyl compounds including E-2 octenal, E-2-nonenal, E,Z-2,6-nonadienal and E,E-2,4-decadienal in beer was developed using stir bar sorptive extraction (SBSE) with in-situ derivatization followed by thermal desorption-GC-MS analysis. The derivatization conditions with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine and the SBSE conditions--sampling mode, salt addition, sample volume, polydimethylsiloxane volume (sample/polydimethylsiloxane phase ratio) and extraction time--were examined. The method showed good linearity over the concentration range from 0.1 to 10 ng ml( 1) for all analytes and the correlation coefficients were higher than 0.9993. The limits of detection ranged from 0.021 to 0.032 ng ml(-1) for all analytes. The recoveries (98-101%) and precision (RSD 2.4-7.3%) of the method were examined by analyzing beer samples fortified at the 0.5-ng ml(-1) level. The method was successfully applied to low-level concentration samples. PMID- 12585328 TI - Conductive polymers as new media for solid-phase extraction: isolation of chlorophenols from water sample. AB - Three different conductive polymers, poly-N-methylaniline (PNMA), polyaniline and polydiphenylamine were synthesised and used as sorbents for the solid-phase extraction of some phenolic compounds from water. The separation and determination was, subsequently, performed by gas chromatography-electron-capture and flame ionization detection. Among these easy-made polymers, PNMA gave better recovery for the extraction of the studied analytes. The capability of the PNMA to extract other non-polar compounds such as chloro- and methylbenzene, decane, decanol, cyclohexane and cyclohexanol was also studied. The polymer showed some sort of selectivity towards aromatic compounds than aliphatic. Preconcentration of sample volumes up to 1 l at pH 2 using 120 mg of PNMA led to acceptable recoveries for phenolic compounds, except for phenol. Recovery of phenol for 100 ml of water was 72%. Limits of detection for chlorophenols when 250 ml river water was preconcentrated were between 1 and 40 ng l(-1), analyzed by GC-electron capture detection. PMID- 12585329 TI - Development and validation of a gas chromatographic-mass spectrometric method for simultaneous identification and quantification of marker compounds including bilobalide, ginkgolides and flavonoids in Ginkgo biloba L. extract and pharmaceutical preparations. AB - A gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the simultaneous determination of seven major chemical markers (bilobalide, ginkgolides A, B, C, kaempferol, quercetin and isorhamnetin) in phytopharmaceuticals of Ginkgo biloba L. The intra-day relative standard deviations (RSD) and inter-day RSD's were based on the analysis of the standardized Ginkgo biloba L. extract on the same day and on the following 3 consecutive days. The intra-day RSD's ranged from 1.21% (bilobalide) to 6.20% (kaempferol). The inter-day RSD's ranged from 2.10% (bilobalide) to 10.42% (isorhamnetin). Mean recoveries ranged from a low of 63.0 +/- 5.3% (isorhamnetin) to a maximum of 103.5 +/- 6.0% (ginkgolide A). Calibration curves were linear in ranges between 2.73 and 36.36 microg/ml for the markers. Limits of detection ranged from a low of 0.5 microg/ml (bilobalide) to a high of 2.5 microg/ml (quercetin). The limits of quantitation were a low of 1.1 microg/ml (gingkolides A, B, C) to a high of 7.5 microg/ml (kaempferol). The method was applied to a standard extract (>6% total terpenoids and >24% total flavonoids) and six ginkgo capsule phytopharmaceuticals. PMID- 12585330 TI - Anomalous effects of helium head pressure carbon dioxide in supercritical fluid chromatography and extraction. AB - Helium head pressure carbon dioxide cylinders are commonly used to facilitate the delivery of liquid CO2 to supercritical fluid extraction and chromatographic pumps. It is usually tacitly assumed that the helium used to increase the delivery pressure of the CO2 cylinders is completely insoluble in liquid CO2 and thus remains isolated in the head space of the delivery cylinder. This assumption is invalid because up to 5 mol% helium can be entrained in the liquid CO2 delivered from helium head pressure cylinders. Significantly, contamination of liquid CO2 with even small amounts of helium can cause many unforeseen and usually deleterious effects in supercritical fluid chromatography and extraction schemes. The observed anomalies include decreased density of the fluid phase, irreproducible extraction and retention, ghost peaks, and even phase separation within the column or extraction vessel. PMID- 12585331 TI - Separation of triadimefon and triadimenol enantiomers and diastereoisomers by supercritical fluid chromatography. AB - The enantiomeric separation of triadimenol and triadimefon on a Chiralpak AD column using supercritical fluid chromatography, was studied in this work. The effect of different modifiers (methanol, ethanol and 2-propanol) was tested, with methanol and ethanol providing the best results for the enantiomeric separation of the two compounds. The enantioseparation of a mixture of triadimenol and triadimefon (six stereoisomers) was achieved in only 15 min using a gradient of ethanol, 200 bar, 35 degrees C and a flow-rate of 2 ml/min. The separation of triadimenol diastereoisomers on different achiral columns (diol, silica and ODS) was also investigated. In this case, the type of organic modifier to be used depended on the stationary phase, the Spherex Diol being the column that gave the best separation. Using this column, resolutions higher than 3 were obtained in analysis times of 5 min with any of the modifiers checked. PMID- 12585332 TI - Development of a chiral non-aqueous capillary electrophoretic system using the partial filling technique with UV and mass spectrometric detection. AB - A chiral non-aqueous CE system with UV and mass spectrometric detection has been developed. The enantioseparation was promoted by diastereomeric complex (ion pair) formation between the amines (e.g. salbutamol, atenolol) and the chiral selector, (-)-2,3:4,6-di-O-isopropylidene-2-keto-L-gulonic acid [(-)-DIKGA]. Different solvent mixtures were studied, as well as different concentrations of ( )-DIKGA and ammonium acetate in the background electrolyte. A partial filling technique was developed with a selector plug composed of (-)-DIKGA and ammonium acetate in a solvent mixture of methanol and 2-propanol. The separated enantiomers of pronethalol were detected by a Q-TOF MS system equipped with a sheath-flow electrospray ionization interface. PMID- 12585333 TI - Simultaneous determination of water-soluble vitamins in a vitamin-enriched drink by an in-capillary enzyme reaction method. AB - The in-capillary enzyme reaction method was used to determine riboflavin phosphate in a vitamin-enriched drink based on its conversion to riboflavin (vitamin B2) with alkaline phosphatase. Simultaneously, three water-soluble vitamins [thiamine nitrate (vitamin B2 mononitrate), pyridoxine hydrochloride (vitamin B6 hydrochloride) and nicotinamide (vitamin PP)] and anhydrous caffeine in the drink were subjected to quantitative analysis. In the system, electrophoretic migration was used to mix zones containing the substrate (riboflavin phosphate) and the enzyme (alkaline phosphatase). The reaction was then allowed to proceed in the presence of a weak electric field and, finally, the product (riboflavin) of enzyme reaction and other water-soluble vitamins migrated under the influence of an applied electric field to the detector. All the active ingredients and the formulation excipients were successfully separated by micellar electrokinetic chromatography with 135 mM sodium dodecyl sulfate. To prevent inhibition of enzyme reaction by the addition of sodium dodecyl sulfate to the reaction zone, sandwich mode injection, in which plugs of sandwich solution without sodium dodecyl sulfate were introduced into the capillary on both sides of the reaction zone, was utilized as a barrier to protect the enzyme reaction from the inhibitor. The relationship between the peak area of the product and the concentration of the substrate was calculated in the in-capillary enzyme reaction method. Excellent linearity was obtained, with correlation coefficients of 0.9999. The established method was validated and demonstrated to be applicable to the determination of the five active ingredients, including riboflavin phosphate, in a commercial vitamin-enriched drink. No interference from the formulation excipients was observed. Good linearities were obtained, with correlation coefficients above 0.999. Recoveries and precisions ranged from 99.3 to 101.8%, and from 0.1 to 2.5% RSD, respectively. Good agreement was obtained between the established method and traditional high-performance liquid chromatographic methods. These results suggest that the in-capillary enzyme reaction method can be used for the simultaneous determination of riboflavin phosphate and other water-soluble vitamins in pharmaceuticals. PMID- 12585334 TI - Separation and determination of phenylpropanoid glycosides from Pedicularis species by capillary electrophoresis. AB - Capillary zone electrophoresis, using 30 mM borate buffer (pH 9.00) with 10% (v/v) methanol, was established for the identification and determination of four phenylpropanoid glycosides (PPGs)--echinocoside (ECH), verbascoside (VER), pedicularioside M (PED-M) and pedicularioside A (PED-A)--in extracts of Pedicularis longiflora var tubiformis, Pedicularis longiflora and Pedicularis Kansuensis. Regression equations revealed linear relationships (correlation coefficients: 0.9993-0.9999) between the peak area of each compound (ECH, VER, PED-M and PED-A) and its concentration. The relative standard deviations of the migration times and peak areas were <1.93 and 4.54%, respectively. The recoveries of four PPGs ranged between 95.6 and 108.4%. The effects of several CE parameters on the resolutions were studied systematically. PMID- 12585335 TI - Glucotoxicity and beta-cell failure in type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is increasing worldwide with a trend of declining age of onset. It is characterized by insulin resistance and a progressive loss of beta cell function. The ability to secrete adequate amounts of insulin is determined by the functional integrity of beta-cells and their overall mass. Glucose, the main regulator of insulin secretion and production, exerts negative effects on beta-cell function when present in excessive amounts over a prolonged period. The multiple metabolic aberrations induced by chronic hyperglycemia in the beta-cell include increased sensitivity to glucose, increased basal insulin release, reduced response to stimulus to secrete insulin, and a gradual depletion of insulin stores. Inadequate insulin production during chronic hyperglycemia results from decreased insulin gene transcription due to hyperglycemia-induced changes in the activity of beta-cell specific transcription factors. Hyperglycemia may negatively affect beta-cell mass by inducing apoptosis without a compensatory increase in beta-cell proliferation and neogenesis. The detrimental effect of excessive glucose concentrations is referred to as 'glucotoxicity'. The present review discusses the role of glucotoxicity in beta cell dysfunction in type 2 diabetes mellitus. PMID- 12585336 TI - How should we manage growth hormone deficiency in adolescence? Transition from paediatric to adult care. PMID- 12585337 TI - Physical repercussions of childhood-onset growth hormone (GH) deficiency and hGH treatment in adulthood. AB - The aim of this study was to evaluate the physical condition of young adults with childhood-onset growth hormone deficiency (GHD) before and after 6 months of hGH therapy. Ten men and three women, aged 22.3 +/- 3.3 years, previously treated with hGH for 8.6 +/- 4.07 years at a dose of 0.5 IU/kg/week with a minimun of 2.5 years without treatment at the time of study, were studied. Nine patients presented isolated GHD and four patients had combined pituitary hormone deficiencies; they were treated with hGH at a dosage of 0.125 IU/kg/week for the first month and 0.25 IU/kg/week for the following 5 months. The tests performed were: exercise test, heart rate, rating of perceived exertion, blood lactate analysis, jump test and hand grip. Body composition was also analyzed using Holtain Body Analysis. Skinfold thickness was measured at four sites (triceps, biceps, subscapular and suprailiac). After 6 months of treatment a significant increase in lean body mass (42.0 +/- 7.72 to 46.2 +/- 8.01 kg, p = 0.004) and decrease in fat mass (19.6 +/- 10.01 to 16.1 +/- 10.79 kg, p = 0.01) were observed. The initial physical condition of these patients was lower than expected, and improved after treatment with an increase in maximum oxygen consumption from 2.0 +/- 1.2 to 2.33 +/- 0.68 l x min(-1) (p = 0.01). Maximum heart rate increased significantly from 189 +/- 14.8 to 193 +/- 11.7 beats x min( 1) (p = 0.03). No modifications were observed in anaerobic threshold (4 mmol x l( 1)). Only slight, non-significant increases were observed in jump and strength tests. We conclude that a) adults with childhood-onset growth hormone deficiency present a deficient physical condition and lower than expected for age and sex; b) this condition improves after 6 months of treatment, particularly in the aerobic aspect; c) changes observed in strength tests were discrete and of little significance; and d) the increase observed in lean body mass plays an important role in these changes. Further studies investigating GH action on maximum oxygen consumption are required, once its basic mechanism of action has been determined, either in the heart or peripheral factors. PMID- 12585338 TI - Effect of hypothyroidism and its treatment on the IGF system in infants and children. AB - We performed a study on infants and children with hypothyroidism to determine the effect of hypothyroidism and its correction on components of the IGF system. A total of 35 patients were subdivided into four groups based on age and severity of the disease. Serum concentrations of immunoreactive IGF-I, free IGF-I, IGFBP-2 and IGFBP-3 were measured before and after treatment and compared to controls matched for age, sex and puberty. Baseline total IGF-I (TIGF-I) concentrations were significantly lower prior to treatment in the infants with severe hypothyroidism and increased significantly after thyroxine therapy. Baseline free IGF-I (FIGF-I) concentration was significantly lower prior to treatment in infants with severe hypothyroidism when compared to controls but did not increase significantly after treatment. In infants with severe and compensated hypothyroidism, IGFBP-3 concentrations prior to treatment were lower when compared to controls. These concentrations increased during treatment. Baseline IGFBP-2 levels did not differ from the control values in both these groups but decreased significantly after correction of the hypothyroidism. Although these changes appeared to occur with thyroxine therapy, multiple regression analysis suggested that age was a more important determinant of the changes observed in these parameters than serum thyroxine concentration. In children with acquired hypothyroidism no difference in any of these parameters was noted between hypothyroid patients and controls. TIGF-I increased significantly on thyroxine therapy, but the difference was small. No significant differences were noted in other measured parameters with thyroxine therapy. In older children with compensated hypothyroidism no significant differences were noted in any of the measured parameters in the pretreatment, post-treatment and control groups. In conclusion, although changes appear in TIGF-I, IGFBP-3 and IGFBP-2 in infants with congenital hypothyroidism when they are treated with thyroxine, age appears to be the more important determinant of these changes than does thyroxine concentration. In older children with acquired hypothyroidism, TIGF-I and FIGF-I levels were not significantly lower than in age- and sex-matched controls. After treatment only TIGF-I levels increased. PMID- 12585339 TI - Bone ultrasound velocity curves of newly born term and preterm infants. AB - BACKGROUND: Ultrasound velocity (speed-of-sound [SOS]) has been proposed as a non invasive method of evaluation of bone status in infants. We hypothesized that SOS correlates with both gestational age and birth weight. METHODS: We measured SOS within the first 96 hours of life at the right tibial midshaft location in 73 neonates ranging in gestational age from 25 to 41 weeks, and in birth weight from 825 to 3880 grams. We used the Sunlight Omnisense 7000p device (Tel Aviv, Israel). Results are expressed as meanS +/- 1 SD; statistical analyses included linear regression and computation of 95% CI regression lines; p<0.05 was considered significant. RESULTS: There was, as hypothesized, a significant correlation between gestational age (or birth weight) and SOS. There were no significant differences between males and females. Ninety-five percent confidence intervals were established. CONCLUSIONS: These data may be used as reference ranges for further studies. PMID- 12585340 TI - Hypothalamic-pituitary-adrenal axis and interleukin-6 activity in children with head trauma and syndrome of inappropriate secretion of antidiuretic hormone. AB - OBJECTIVE: Arginine vasopressin (AVP; anti-diuretic hormone) and corticotropin releasing hormone are the two major secretagogues of hypophyseal adrenocorticotropin (ACTH). Interleukin-6 (IL-6) is a potent stimulator of the human hypothalamic-pituitary-adrenal axis (HPA) and a secretagogue of both parvocellular and magnocelullar AVP. We have previously suggested that IL-6 stimulated AVP secretion may be the origin of the syndrome of inappropriate antidiuretic hormone secretion (SIADH) in inflammatory conditions. STUDY DESIGN: To further elucidate the relationship between IL-6 and SIADH as well as IL-6 and HPA axis activity, we studied eight previously healthy children (age 6.3 +/- 4.8 [mean +/- SD] years, weight 23.8 +/- 10.5 kg) who--after sustaining head trauma- presented SIADH during hospitalization (for 7.1 +/- 3.8 days) in the pediatric intensive care unit (P-ICU). Routine blood samples were taken twice daily at 08:00 and 20:00 hours. All children but one survived. Measurements included blood and urine osmolality (BlOsm and UrOsm, respectively), serum cortisol (F) and IL 6, plasma ACTH and AVP. Correlations were assessed with linear regression among the areas under the curve (AUC) of BlOsm, UrOsm, ACTH, F, IL-6 and AVP, separately for BlOsm values <280 mOsm (SIADH phase) and > or = 280 mOsm (non SIADH phase). RESULTS: During the SIADH phase AVP-AUC correlated positively with IL-6-AUC (r = +0.96, p<0.05), BlOsm-AUC correlated positively with AVP-AUC and F AUC (r = +0.95 and +0.98, p<0.05, respectively) and F-AUC correlated positively with ACTH-AUC (r = +0.99, p<0.05). During the non-SIADH phase ACTH-AUC correlated positively with BlOsm-AUC (r = +0.96, p<0.05). CONCLUSION: IL-6 secreted during an aseptic inflammatory state, such as sustaining head trauma with SIADH, is quantitatively correlated to AVP, indicating that this cytokine is directly and/or indirectly involved in the pathogenesis of SIADH. PMID- 12585341 TI - Effect of low-dose testosterone treatment on androgen regulated proteins prostate specific antigen and sex hormone binding globulin in short prepubertal boys: lack of initiation of puberty. AB - The efficacy of testosterone undecanoate (TU) treatment in constitutional delay of growth (CHD) is well recognized. We investigated its role in initiating puberty. Sera taken prior to, just after 6 months on and after 6 months off treatment with TU (20 mg daily) were analyzed from eight boys and compared to results from eight boys receiving placebo. Prostate specific antigen (PSA) and sex hormone binding globulin (SHBG), sleep-entrained pulsatility and mean overnight luteinizing hormone (mLH), and morning testosterone (T) levels were measured. Free androgen index (FAI) was calculated. Testicular volume (TV) and growth parameters were assessed. During treatment, there was a significant increase in height velocity in boys taking TU vs placebo (mean +/- SD: 5.7 +/- 2.0 vs 3.2 +/- 0.9 cm/year, p = 0.008) but no significant differences were observed in regard to LH pulsatility, mLH, T, SHBG, FAI, PSA and TV values. PSA was detectable in four patients (two each in the TU and placebo groups) at 6 months off treatment indicating pubertal progression. Among the hormones measured, only pretreatment mLH levels were significantly higher in the PSA positive patients compared to 12 PSA-negative patients (mean +/- SEM: 1.5 +/- 0.39 vs 0.37 +/- 0.06 IU/l, p < 0.001). In conclusion, TU treatment shows no significant effect on initiation or advancement of puberty despite its resultant growth acceleration. Among the hormonal changes studied, mLH levels were the earliest indicator of pubertal initiation. PMID- 12585342 TI - Analysis of changes in the percentage of B (CD19) and T (CD3) lymphocytes, subsets CD4, CD8 and their memory (CD45RO), and naive (CD45RA) T cells in children with immune and non-immune thyroid diseases. AB - Graves' disease (GD) is an autoimmune thyroid disease caused by immunological abnormality. The immune cells (lymphocytes T and B) which infiltrate the thyroid gland play a key role in the development of autoimmune thyroid disease (AITD). The aim of this study was to evaluate the differences between distribution of T (CD3) lymphocytes, subsets CD4, CD8, and their memory (CD45RO), and naive (CD45RA) T cells and B (CD19) lymphocytes in the peripheral blood of patients with Graves' disease (GD) (n = 33, mean age 15.9 +/- 5.9 years) and non-toxic nodular goiter (NTNG) (n = 25, mean age 15.2 years), in comparison to age- and sexmatched healthy control subjects (n = 25, mean age 15.9 years). The percentages of peripheral blood lymphocyte subsets were analyzed by three-color flow cytometry using a Coulter EPICS XL cytometer. In the untreated Graves' patients we observed an increase in the percentage of CD19+ (p<0.007, p<0.003), CD4+ (p<0.004, p<0.017), CD4+CD45RO+ (p<0.04, NS), CD4/CD8 ratio (p<0.002, p<0.001) and a decrease in the percentage of CD8+ (p<0.02, p<0.02), CD4+CD45RA+ (p<0.04, p<0.03) cells in comparison to the healthy control subjects and euthyroid Graves' patients. These abnormalities were absent in children with non toxic nodular goiter. In addition, the levels of CD3+, CD4+CD8+, CD8+CD45RO+ T cells and CD8 lymphocytes co-expressing CD45RA and CD45RO antigens were similar in all groups and no statistically significant differences were found in comparison to the healthy controls. In the untreated Graves' patients we found a positive correlation between serum levels of fT4 and fT3 and the percentage of CD19+ lymphocytes (r = 0.45, p<0.01, r = 0.37, p<0.04), between serum level of fT4 and the percentage of CD4CD45RO (r = 0.4, p<0.02) lymphocytes and between concentration of TRAb and CD4+ (r = 0.38, p <0.04) and CD19+ (r = 0.39, p<0.016) cells. Statistically significant negative correlations existed between TRAb, TPO Ab or TG-Ab concentration in blood serum and the percentage of CD8+ lymphocytes (r = -0.55, p<0.002; r = -0.41, p<0.02; r = -0.51, p<0.004), and between fT4 concentration and the percentage of CD8+ (r = -0.39, p<0.02) lymphocytes. No such correlation was detected in patients with non-toxic nodular goiter. We conclude that the abnormal distribution of B lymphocytes, memory and naive T cell subsets in the peripheral blood in children and adolescents with untreated Graves' disease suggests their role in the development of autoimmunity. The normalization in the percentage of these immune cells after thyrostatic treatment in comparison to newly diagnosed patients confirms the immunomodulatory effect of methimazole therapy. PMID- 12585343 TI - Increasing prevalence of type 2 diabetes mellitus in Thai children and adolescents associated with increasing prevalence of obesity. AB - Type 2 diabetes mellitus (DM) is being diagnosed more frequently in children and adolescents. Thailand has a low incidence of childhood DM. This study reviewed patients with DM in the Division of Pediatric Endocrinology, Faculty of Medicine, Siriraj Hospital compared to our previous study. The results demonstrate that type 2 DM in Thai children and adolescents has increased from 5% during 1986-1995 to 17.9% during 1996-1999. Mean age was 11.6 years. Mean BMI was 27.8 kg/m2. Fifty-six percent were diagnosed on routine examination. The period of increase in type 2 DM is associated with an increase of obesity prevalence from 5.8% in 1990 to 13.3% in 1996. This result emphasizes the importance of encouraging daily physical activity and healthy diet in our populations and also alerts our pediatricians and endocrinologists to the possibilities of type 2 DM in these age groups. PMID- 12585344 TI - Coexistence of impaired indices of autonomic neuropathy and diabetic nephropathy in a cohort of children with type 1 diabetes mellitus. AB - One hundred and twenty-nine (87%) out of a county population of 150 eligible children with type 1 diabetes mellitus (DM) and 144 age- and sex-matched control children participated in a longitudinal, epidemiological study of the evolution of diabetic microvascular disease. The mean (SD) age of the children with DM was 12.4 (+/- 3.4) years with a mean DM duration of 4.5 (+/- 3.2) years and a mean HbA1c of 11.1 (+/- 2.2)%. Two sets of measurements were made over a period of 18 months. Pupillary adaptation in darkness (PD), as an index of sympathetic dysfunction, was assessed using a portable Polaroid pupillometer. Urinary albumin excretion, as an index of incipient nephropathy, was assessed in children with DM from a fraction of all voidings during two separate 48-h periods and was expressed as urinary albumin/creatinine ratio (A/C). Heart rate variation (HRV), as an index of cardiovascular autonomic dysfunction, was assessed using a heart rate monitor and computer. Blood pressure (BP) was measured using a random zero sphygmomanometer. Reduced PD was found in 14/129 (13.8%) children with DM and in 5.8% of the controls. The diabetic children with reduced PD had longer DM duration, higher HbA1c, raised diastolic BP and higher urinary A/C than those with normal PD. Fifty percent of these children also had increased A/C and impaired HRV, in addition to impaired PD. Another group of 25/129 (19.4%) children with DM presented intermittently raised mean urinary A/C, while five children presented persistently raised A/C in both 48-h urine collections. The characteristics which differentiated the microalbuminuric group from the normoalbuminuric one were older age, longer DM duration, poorer glycemic control, reduced PD, impaired HRV and raised diastolic BP. It seems that diabetic nephropathy (DN) and autonomic neuropathy (DAN) exist in concert in childhood DM. Diabetic children with impaired indices of DN and DAN are older, pubertal and have longer DM duration and higher HbA1c values than the rest of the diabetic population. PMID- 12585345 TI - Estradiol levels in girls with Turner's syndrome compared to normal prepubertal girls as determined by an ultrasensitive assay. AB - Based on growing evidence that estradiol is produced in small amounts even in the prepubertal ovary, we hypothesized that estradiol levels in girls with Turner's syndrome (TS) are lower than in normal prepubertal girls secondary to the lack of normally functioning ovaries. Estradiol levels in untreated girls with TS have not been previously well defined because of the lack of adequate sensitivity of previously available estradiol assays. We utilized an ultrasensitive assay to study estradiol levels in 34 girls with TS and 34 normal age-matched prepubertal girls between the ages of 5 and 12 years. The average estradiol level in the girls with TS (6.4 +/- 4.9 pmol/l estradiol equivalents) was significantly lower than in the normal prepubertal girls (12.7 +/- 10.8 pmol/l estradiol equivalents; p < 0.01). Girls with TS were significantly shorter, and weighed less than the normal prepubertal girls, as expected. The estradiol level was not significantly correlated with height, bone age, or degree of bone age delay. In conclusion, girls with TS have significantly lower estradiol levels than normal age-matched prepubertal girls. This report is consistent with the hypothesis that the lack of normal ovarian function in girls with TS is evident even before puberty. PMID- 12585347 TI - Polyglandular autoimmune syndrome type 1 without chronic mucocutaneous candidiasis in a 16 year-old male. AB - A 16 year-old boy presented with adrenal crisis and was incidentally found to have features of latent tetany. Also detected were Hashimoto's thyroiditis, alopecia and subnormal T cell function, and he was diagnosed as having polyglandular autoimmune syndrome type 1 (PGA-1), although chronic mucocutaneous candidiasis, a hallmark of PGA-1, was absent. The presentation of several components of this disorder at one time and at this age is uncommon, and the features of overt hypoparathyroidism were probably masked by associated adrenal insufficiency. PMID- 12585346 TI - Malignant astrocytoma arising 10 years after combined treatment of craniopharyngioma. AB - Craniopharyngioma is the third most common intracranial tumor in childhood. Following surgery, virtually all patients present with hypopituitarism and are at considerable risk of tumor recurrence. Secondary tumors, however, are rare, occurring usually 10 years after diagnosis and associated with poor prognosis. We report on a 5 year-old boy in whom craniopharyngioma was diagnosed due to unilateral visual loss. After surgery he underwent conventional radiation therapy with a total tumor dose of 55 Gy, and had hormonal support with DDAVP, thyroxine, and a variable dose of hydrocortisone. Growth velocity declined slowly in the first 4 years, but improved later on again without GH therapy despite abnormal provocative tests. At the age of 15 years he developed peripheral facial nerve palsy due to a malignant astrocytoma (WHO grade III/IV). Repeated conventional radiation therapy with an additional stereotactic boost and chemotherapy could not prevent the fatal outcome. This observation may temper the use of radiosurgery in benign intracranial tumors. PMID- 12585348 TI - Precocious puberty secondary to topical testosterone exposure. AB - We report a case of pronounced virilization, including marked penile and pubic hair growth, accelerated height velocity and skeletal maturation, and increased muscle mass in a 2.67 year-old boy resulting from presumed inadvertent, long-term exposure to a topical testosterone cream being used by his father. PMID- 12585350 TI - The role of surgery in the treatment of carcinoma of the pancreatic head. AB - Surgery in pancreatic carcinoma has changed within the last two decades. One of two patients nowadays can be treated without operation. Palliative surgical procedures rarely have to be done. Actually jaundice in patients with unresectable pancreatic cancer is treated by stenting instead of performing the classic biliodigestive anastomosis. In our department the rate of patients suitable for radical surgery lays constantly between 20 and 25% within the last decades. However, this high percentage of potentially curable patients may be due to a "positive patient selection" referred to a University hospital. In accordance with international publications it seems to be certain, that surgical therapy for pancreatic cancer has reached its limitations. Due to these open questions, working on pancreatic cancer will remain one of the most challenging but worthwhile features for generations of surgeons. (Tab. 4, Fig. 7, Ref. 16). PMID- 12585349 TI - Transient neonatal hypothyroidism due to a maternal vegan diet. AB - Iodine is an important constituent of thyroid hormones and deficiency can lead to a range of problems depending on the degree and at what stage of life the deficiency occurs. We report a 10 day-old infant with a goitre, who presented with raised TSH on dried blood spot screening. It was observed that her mother also had a goitre. The mother was a vegan and, on dietary assessment, her iodine intake was extremely low. Both mother and infant had abnormal thyroid function tests. Mother was given Lugol's iodine and her thyroid function tests normalised. Her baby was initially prescribed thyroxine on the basis of the raised screening TSH. This was subsequently withdrawn at the age of 2 weeks, following a normal plasma TSH. Thyroid function tests remained normal and the goitre disappeared by the age of 2 months. Iodine deficiency is uncommon in the Western World. However the incidence may be rising in otherwise iodine replete areas, particularly in those who adhere to restrictive and unusual diets. In the case of pregnant mothers their unborn child's health is in danger. This report demonstrates the need to ascertain maternal diets early in antenatal care, and supplement if necessary to avoid risk to their own health and that of their offspring. PMID- 12585351 TI - Radical surgery for cancer of the pancreas. AB - The basic principles of oncological radicality in surgery are: a) the surgeon have to perform always an R0 resection and he have to use perioperative investigation of resection margin, b) lymphadenectomy must be rational meaning that it is enough to perform regional lymphadenectomy (radical lymphadenectomy is not necessary), c) if portal vein resection enables increased curability of resection, then it is valuable to resect part of this vessel and to perform end to-end venous anastomosis. (Ref. 5.). PMID- 12585352 TI - Surgery of the tumours of the liver. AB - BACKGROUND: Recent decades have seen new methods of resection techniques which have contributed to improved results. AIM: To provide a review of state-of-the art knowledge of surgical treatment of liver tumours. The clinical material is analyzed. METHODS: Retrospective analysis of the patients of clinical department of surgery of the university hospital. A representative group of patients could be studied in years 1998-2001 when the ultrasonic (harmonic) scalpel became available for liver resection in our patients. RESULTS: The study consists of 99 patients with an average age of 59 years. Haemangioma was the most common benign tumour (7 cases) and hepatocellular carcinoma was the most frequent primary malignancy (18 cases). The prevailing cause among secondary malignant liver masses was metastasis of colorectal carcinoma (20 cases). The most frequently used anatomical resections were right hemihepatectomy or left hemihepatectomy and bisegmentectomy in 18 and 14 patients respectively. Local complications occurred in 17% and systemic complications in 6% of patients. There were no postoperative deaths. CONCLUSION: Liver resection, either anatomical or non anatomical, remains the golden standard among the methods of treatment of hepatic tumours. New techniques have contributed to the traditional hepatic resection. The authors' group showed promising short term results. (Tab. 7, Ref. 31.). PMID- 12585353 TI - Oncological radicality in colonic cancer operation. AB - Lymphadectomy should extend to the level of the origin of the primary feeding vessel, suspected positive lymph nodes outside the standart resection should be removed when feasible. A minimum 5 cm bowel margins proximally and distally should be used and peroperative histology must confermend tumor free anastomosis. En-block resection should be performed for tumors adherent to local structures. Inadvertent bowel perforation increases the risk of recurrence and should be avoided. Ovaries grossly involved with tumor should be removed. In all women having operations for cancer of the colon bilateral oophorectomy should be performed. In premenopausal women the ovary is a frequent site for metastatic disease (the Krukenberg tumor) which occures as often as 25 percent of the time. The incidence of of ovarian metastasis is much lower in postmenopausal women but the consequences of oophorectomy are also less severe in that age group. (Ref. 9.). PMID- 12585354 TI - Tumors of the rectum and anus. AB - Within a ten year period we have performed 433 radical operations on the rectum. An anterior resection of rectum was done in 110 cases, low anterior resection in 60 patients, very low anterior resection was done by stapler in 32 patients and was sewn by hand in 10 patients. Colo-anal anastomoses were performed 86 times and abdominoperineal amputation of rectum sec. Miles 20 times. Local transsphincteric procedures were used 14 times. In 12 patients the colo-anal anastomosis was performed either for recurrence of tumor or after a Hartmann's procedure. In the carcinoma of the lower half of the rectum there is still an unsolved question: whether to perform a low or a very deep low anastomosis or whether an extirpation of rectum with sparing the sphincters and sewing a colo anal anastomosis is more beneficial. In the years 1980-1995 we treated surgically 64 patients with a tumor in anal area. All patient underwent preoperative radio- and chemotherapy. In our group adenocarcinoma was demonstrated in 6 patients, squamous carcinoma in 48 patients and squamous cells of the carcinoma in situ in 10 patients. PMID- 12585355 TI - Reconstruction after total gastrectomy. AB - After total gastrectomy it is necessary to realize the reconstruction of the upper part digestive tract. It is necessary to choose the reconstruction method, which provides good life quality, and extensive physiologic regulation of gastrointestinal hormones after total gastrectomy. There is no ideal reconstruction method. Nowadays there are about 70 kinds of reconstruction methods. It is very important to preserve the duodenal passage. Also the production of reservoir is important. The conclusions of studies are not homogeneous. The patients can benefit from the reconstruction preserving duodenal passage and formed by pouch. (Fig. 1, Ref. 23.). PMID- 12585356 TI - Familial adenomatous polyposis as a precancerosis of colon cancer. AB - Familial adenomatous polyposis is a genetic disorder caused by mutations of the adenomatous polyposis coli gene. This gene is localized on chromosome 5q21. The incidence of the disease is 1/7000-8000. Surgery has an important role in the management of familial adenomatous polyposis. In the absence of prophylactic colectomy, death from colom cancer will occur in virtually all of familial adenomatous polyposiscases by age 50, with 37% affected by colon cancer by age 37. (Ref. 8). PMID- 12585357 TI - Chemodectoma--carotid body tumor surgical treatment. AB - Chemodectoma is a rare tumor of the neuroendocrinne tissue of the carotid body. In the most of cases is benign (noncancerous), but can have a malignant behave. The authors describe their experience about the surgical treatment of the chemodectoma in years 1995-2001. We treated two cases of this tumor in our department during the period of 6 years. The only one preferred method of care is the surgical removing. Radiation therapy may also be used. (Ref. 11.). PMID- 12585358 TI - Radical D2 surgery for patients with gastric cancer. AB - The authors analyzed and prepared a report concerning 18 radical surgeries for gastric cancer that were performed between 1999-2001. Overall, 55 operation were performed, 32 radical, 18 palliative and 5 explorative laparatomies. D2 resections were performed 18 times, while D1 type 14 times. The group undergoing D2 surgery comprised of 10 men and 8 women with average age of 64.3. D2 resection included partial (8 times), or total (10) gastrectomy and lymfadenectomy of perigastric nodes, supra and infrapyloric nodes and nodes along common hepatic artery, truncus coeliacus, lienal artery, left paracardial nodes and removing capsula of pancreas. Splenectomy was performed twice. On average, 37.5 lymphnodes were removed for every operation (25-69). Operative mortality was none (0%) and morbidity was 22%. As of January 1, 2002 relaps was noted in six patients, and 5 patients died. CONCLUSION: D2 resection surgeries performed by an experienced surgeon show low morbidity as well as better outcome and higher perspective for long-term survival in patients with gastric cancer. (Tab. 1, Ref. 19.). PMID- 12585360 TI - Significance of a prolonged postoperative bowel paralysis in complete dehiscence of laparotomy wound. AB - We performed 910 laparotomies in the years 1998-1999. Complete dehiscence of a laparotomy wound was observed in 18 cases. We compare the duration of postoperative bowel paralysis in patients with and without eventration. The statistical comparison of the two groups was performed with the Wilcoxon test, the level of significance was set to 5%. We have found that the duration of postoperative bowel paralysis was significantly longer in the group of patients with eventration. (Tab. 4, Ref. 5.). PMID- 12585359 TI - CEA and relapse after the operation of colorectal carcinoma. AB - BACKGROUND: Relapses have an important meaning in relation to the curative surgical intervention. In RO resections according to UICC classification, the local relapses were classified as the most important factor for survival. Without any doubts the most important of these tumor-biological prognostic factors in patients with colorectal carcinoma is the carcino-embryonal antigen. Up to now, the unquestionable importance of the determination of serum levels of CEA for the detection of relapses is most often described. MATERIAL AND METHODS: This study is retrospective, not randomized. Analyzed were medical records and data of patients, who underwent the surgery of colorectal carcinoma at IInd Department of Surgery, University Hospital, Faculty of Medicine, Comenius University, Bratislava, in period from January 1st, 1986 to December 31st, 1995. In our analysis we evaluate the age and gender of patients, date of the surgery and type of the surgery from the point of acuteness, as well as radicality, staging of the disease, levels of the serum CEA. Serum levels of carcino-embryonal antigen before operation were determined and evaluated. In patients, who remaind in the monitoring at our department we examine CEA in regular intervals. RESULTS: From our group of patients, in 28 patients the relapse of the disease was determined here. Before the first operation, the preoperative level of CEA was examined in 15 patients and in 13 patients it was not. From 15 patients with examined CEA level, 7 patients (46.7%) had increased and 9 patients (53.3%) normal level. In twenty-seven patients CEA level was examined to reveal a relapse. One patient was diagnosed during the acute surgery and CEA was not determined prior this operation. From the remaining 27 patients, CEA level was positive in 20 patients (74.1%) and 7 patients had normal levels (25.9%). From these 7 patients with normal CEA level, in 5 cases it was locoregional relapse and 2 patients have distant metastases. From all patients, in whom the relapse was determined at our department, in 13 patients locoregional relapse was detected and in 15 patients distant metastases were diagnosed. CONCLUSION: Due to the sensitivity of serum level of CEA for detection of relapses, which is higher compared to the primary tumor and also to found lead time 4 months we think the postoperative regular determination of CEA in patients after the curative resection is appropriate. Earlier determination of less progressive relapse is definitely important, although the current long-term survival after the surgery for relapses is not optimistic. Development of surgical procedures as well as alternative treatment can bring better results in less developed relapse of the disease. (Ref. 28.). PMID- 12585361 TI - Limited versus total stripping of vena saphena magna. AB - BACKGROUND: There are two different ways to operate on patients with varicose veins. OBJECTIVES: Prospective evaluation of two groups of patients operated on in two different ways to find out more convenient procedure for varicose veins surgery. METHODS: The results of 577 patients operated on for lower limbs varicose veins were evaluated. The patients were divided into two groups. The first group comprised of 125 patients in whom limited stripping of vena saphena magna was performed. The second group comprised of 397 patients in whom total stripping of vena saphena magna was applied. In the remaining 55 patients an operation other than VSM was performed in the venous system of lower limbs. During the follow-up three months after the operation the main attention was paid to a possible neurological disturbance. RESULTS: Much better results were observed in the group of patients treated with the method of limited stripping. Neurological disturbances occurred in two patients only (i.e. 1.6%). Within the other group disturbances were encountered in 28 patients (i.e. 7%). CONCLUSIONS: Limited stripping of vena saphena magna results in lower number of neurological disturbances. (Tab. 1, Ref. 10.). PMID- 12585362 TI - Renal autotransplantation in irreversible ureteral injury. AB - The authors describe a case of ureteral avulsion in ureteroscopy and extraction of calculus from the central part of the ureter. They chose a renal autotransplantation for the ureter reconstruction. They share the same opinion as the authors who use renal autotransplantation as the method of choice in irreversible ureteral injury. Ureteral avulsion is the most severe complication of ureteroscopy. The authors described a case of right ureteral avulsion, with subsequent autotransplantation as the final solution. (Fig. 6, Ref. 7.). PMID- 12585363 TI - Iatrogenic esophageal perforation--diagnosis and therapy. AB - The diagnosis and therapy of iatrogenic esophageal perforation in a group of patients treated for this condition. During the period of 1994-2002 twelve patients suffered iatrogenic esophageal perforation with a mortality rate of 33.3%. The therapeutical approach consisted of either primary repair of the perforation, drainage of the mediastinum and pleural cavity or inserting a stent. (Ref. 6.). PMID- 12585364 TI - The treatment of venous leg ulcers by compression sclerotherapy. AB - PURPOSE: To assess the results of complex sclerotization therapy of pure venous ulcers on all levels of venous system. PATIENTS: Over the last 10 years the authors treated 698 patients with venous ulcerations and compared the therapeutic results in 3 groups of patients treated by Fegan's technique of compression sclerotherapy combined with an antireflux operation of the deep venous system in ulcers not responding to superficial and perforator therapy. RESULTS: 84% of patients with venous ulcers had pathological reflux in the deep venous system and all of them (100%) had insufficiency of the perforators. Using Fegan's technique of compression sclerotherapy the authors managed to heal 95.4% of ulcers with an average time of complete ulcer epithelialization of 39 +/- 12 days. The recurrence rate within 5 years was 18% (126 patients). The average time of disability during the course of sclerotisation was 7 days and no hospitalisation was needed. The healing process depends on the presence of reflux in the popliteal vein in statistically significant fashion. In patients, where a new valve was constructed or direct valvuloplasty was done, all ulcers (except 3) responded to the compression sclerotherapy and healed within 3 months. CONCLUSION: The authors consider compression sclerotherapy (Fegan's technique) to be the easiest and most effective way of treating patients with venous ulcers. Direct valve repair techniques are appropriate only in non-thrombotic deep vein reflux which is very rare (0.6% - 4 patients). In patients with torpid ulcers, where compression sclerotherapy failed (4.6% - 28 patients) in situ construction of a new venous valve helped to heal these ulcers. (Tab. 1, Ref. 18.). PMID- 12585365 TI - Abscesses in the liver: amoebic or pyogenic? PMID- 12585366 TI - Amoebic versus pyogenic liver abscess. AB - OBJECTIVES: To study the differences between the clinical presentation and complications of amoebic and pyogenic liver abscess. To correlate the diagnostic significance of Entamoeba Indirect Haemagglutination test (E.IHA) in establishing the diagnosis of amoebic liver abscess. DESIGN: Open cohort observational study. SETTING: Department of Medicine (Medical Unit II) Jinnah Postgraduate Medical Centre, Karachi. PARTICIPANTS: Fifty two patients aged 13-70 years admitted in Medical Unit II and diagnosed to have liver abscess. INTERVENTIONS: Group A comprised of patients clinically diagnosed to have amoebic liver abscess and received Metronidazole 500mg iv x 8 hourly for atleast 10 days or more if patient developed complications. Group B comprised of patients diagnosed to have pyogenic abscess and received Cefotaxime 1g iv x 8 hourly for the same duration or more if complicated (antibiotic reviewed in accordance with culture and sensitivity report). MAIN OUTCOME MEASURES: Differences in the clinical presentation, complications and diagnostic parameters between the two groups of patients. RESULTS: It was not possible to differentiate between amoebic and pyogenic liver abscess on clinical grounds, routine investigations and imaging techniques. Aspiration of pus, especially if the abscess was multiple, was most helpful in differentiating the two types of abscesses. Serological test of E.IHA was highly specific and sensitive for amoebic liver abscess. CONCLUSION: Majority of liver abscesses in Karachi are due to Entamoeba Histolytica. Pyogenic abscess though less frequent, must be excluded by pus aspiration and culture and sensitivity. E.IHA is a good rapid method of discriminating between the two types of abscesses. PMID- 12585367 TI - Recent trend in the radiological presentation of pulmonary tuberculosis in Pakistani adults. AB - OBJECTIVE: To determine the radiological presentation of pulmonary tuberculosis in adults and to compare it with that of other national and international studies. METHOD: This prospective study was done on 150 newly diagnosed smear positive pulmonary tuberculosis patients. Two chest physicians reviewed the chest radiographs. Those x-rays were selected in which there were no difference of opinion. RESULTS: There were 77 male and 73 females. The average age was 34.52 years with range of 17 to 80 years. Out of 150 films, eighty-seven (58%) showed the typical pattern (infiltration and/or nodules with or without cavitations, involving upper zone). While sixty three (42%) showed the atypical pattern (pattern other than typical one). CONCLUSION: Typical pattern of pulmonary tuberculosis is still the common mode of presentation in adults but there is a trend toward increase in atypical pattern. PMID- 12585368 TI - Correlation of the density of yeast Malassezia with the clinical severity of seborrhoeic dermatitis. AB - OBJECTIVE: To correlate the density of the yeast Malassezia with the clinical seventy of seborrhoeic dermatitis. METHOD: Fifty patients and twenty control subjects were selected for the study. The patients were evaluated both clinically for the severity of seborrhoeic dermatitis and microscopically for the presence of the yeast Malassezia. RESULTS: Out of 50 patients Malassezia was present in 41 patients (82%). On microscopic evaluation it was found that patients with mild seborrhoeic dermatiis had a density of 2+ (more than 5 but less than 10 yeast cells per high power field (hpf). Patients with moderate seborrhoeic dermatitis had a density of 3+ (more than 10 but less than 20 yeast cells per hpf) and patients with severe seborrhoeic dermatitis had a density of 4+ (more than 20 yeast cells per hpf). Of the 20 normal subjects only 8 (40%) had Malassezia and they had a density of 1+ (5 or fewer yeast cells per hpf). The results show a strong correlation of the yeast Malassezia to the severity of seborrhoeic dermatitis (p value < 0.05). CONCLUSION: Malassezia increases in proportion with the severity of seborrhoeic dermatitis; an antifungal agent should therefore be used in the treatment of seborrhoeic dermatitis. PMID- 12585369 TI - Child labour a reality: results from a study of a squatter settlement of Karachi. AB - INTRODUCTION: Pakistan's exports have suffered heavy losses due to its denunciation for use of children in the industry on the premise that it destroys their childhood and desires for progress. The international community has not realized the root cause of child employment and imposed harsh economic sanctions that have harmed Pakistan's economy leading to further deterioration of health and well being of these unprivileged children. OBJECTIVES: To identify the reasons behind child labour and to asses the occupational, health and related problems faced by the working children. METHODS: This study was done in a section of a squatter settlement Bhutta village near the seaport called Accora with a population of 10000 (approx.). It was a cross-sectional study using a structured questionnaire with questions regarding occupation, eating habits, health, hopes and aspirations. Respondents were 150 male working children permanent residents of Bhutta Village (Accora) selected by systematic sampling. RESULTS: The children were between the ages of 12-14 years (mean = 13.91 yrs). The average family size of these children was 7.4 members and the average family income was Pak Rupees 2,884/month. Fifty seven percent said they get no time to rest whereas the remaining got half to one hour of rest during the day. The usual foods consumed included pulses (46%), vegetables (39%) and meat (15%). Eighty seven percent were not suffering from any disease at the time of interview. The children had an average income of Pak Rupees 615/month (range: 200-1200). Forty five percent were helpers in shops the rest were employed in other businesses. The reasons given for job included family support (89%), parent's pressure (5%), fond of work (4%), learning skills (1%) and self support (1%). The majority (56%) was working for more than 7 hours per-day and 79% said that they do not like to work. Majority of the children wanted to study, play, be born in another family and become doctors, engineers and officers. CONCLUSION: The children in this area were deprived and were trapped between work for family and the desire of living the life of a child with hopes dreams and aspiration for a better future. Making them jobless is only going to aggravate their health and financial problems and will not improve their social and economic status. A policy for supporting these children has to be worked out which caters to their need for financial support and at the same time provides them with learning opportunities. PMID- 12585370 TI - Ureteroscopy (URS): an effective interventional and diagnostic modality. AB - OBJECTIVE: To study the effectiveness and complications of the use of ureteroscope in the management of ureteral calculi and as diagnostic modality for ureteric strictures and hydroureter. PATIENTS AND METHOD: URS was carried out in 125 consecutive patients. Technical failure to negotiate the ureteric orifice was observed in 10 (8%) patients. Three of them had passive dilatation with ureteric catheter for 48 hours, while in 7, the failures were due to inability to traverse the ureter. This study will thus review 118 patients, 87 males (73.7%) and 31 females (26.3%) with a mean age 38 years +/- 12 years during 24 months. All the ureteroscopies were performed with a 8 FG rigid ureteroscope under G.A. Swiss (pneumatic) lithoclast was used for lithoripsy. Of the 118 patients, 98 calculi were treated. Five of these patients had associated ureteric strictures. Twenty patients underwent ureterorenoscopy for diagnostic purposes. Ten of them had ureteric strictures and in 10 patients dilated ureters with no luminal obstruction was found. RESULTS: Four (4%) of the stones were located in the upper, 13 (13.2%) in middle and 81 (82.6%) in the lower ureter. The size of the stones treated ranged between 6-22 mm. Clearance of stones within 48 hours and 15 days was observed in 66.3% and 93.8% respectively. Only 6.1% of the patients had failure to clear calculus after 30 days and repeat procedures were required. Four of the patients with ureteric strictures had ureteroneocystostomy while in 6 endoscopic dilatation was done. Thirty one and 10 patients had fever upto 38.5 degrees C and more than 38.5 degrees C respectively. Percutaneous Nephrostomy (PCN) placement was required in 2 patients and stripping of mucosa occurred in 1 patient. Mild hematuira was observed in 46 (38.9%) patients. Only 2.5% had moderate degree of hematuria after the procedure. CONCLUSION: Considering the result from this study we suggest that URS is an effective interventional and diagnostic modality for ureteric diseases with a low complication rate. PMID- 12585371 TI - Prevalence of and factors associated with anxiety and depression among women in a lower middle class semi-urban community of Karachi, Pakistan. AB - OBJECTIVE: To study the prevalence of, and factors associated with anxiety and depression among women. DESIGN: A cross sectional survey. SETTING: A lower middle class semi-urban community of Karachi, Pakistan. PARTICIPANTS: A total of 1218 women between the ages of 18-50 years. METHODOLOGY: Systematically every third household was identified from which a woman was randomly selected. The Aga Khan University Anxiety and Depression Scale and a socio-demographic questionnaire were administered verbally by trained interviewers for assessing the prevalence of, and associated factors for anxiety and depression. RESULTS: A prevalence of 30% was found. Increasing age, lack of education and verbal abuse were the associated factors found to have an independent relationship. CONCLUSION: Providing education and reducing domestic abuse could lead to decrease in the prevalence of anxiety and depression in women. PMID- 12585373 TI - Aplasia cutis congenita and antithyroid drugs. PMID- 12585372 TI - Efficacy and tolerability of Paroxetine 20 mg daily in the treatment of depression and depression associated anxiety. AB - OBJECTIVE: To assess the efficacy and tolerability of paroxetine 20mg daily, for the treatment of depression and depression associated with anxiety. METHOD: An open, non-comparative study undertaken at three centers in three cities of Pakistan. A total of 112 in- or out-patients, presenting with a major depressive episode were included in the study. Depression was diagnosed according to DSM IV criteria and a Hamilton Rating Scale for Depression (HAMD-D) score of >18 on the first 17 items of the HAM-D-21. After a placebo washout period of 7 days, patients were given a 20-mg fixed dose of paroxetine daily in the morning for a period of 6 weeks. After baseline, regular assessments were made at 1, 2, 4 and 6 weeks. Efficacy measures included the HAMD-D, the Clinical Anxiety Scale, The Clinical Global Impression Severity of Illness Scale (CGI-S), and the Clinical Global Impression Improvement Scale (CGI-I). Tolerability was assessed by any adverse event. The primary therapeutic outcome measures were patients who achieved a 50% or greater reduction in HAMD-D score at the end of the treatment or achieved a reduction in final HAMD-D score of < or = 10 points. The final scores of 1 or 2 for CGI-S and CGI-I were considered as full clinical response. RESULTS: Of 112 cases evaluated, 57 were males and 55 females whose ages ranged from 18 to 65 years. There was a clear and statistically significant efficacy of paroxetine on all the major outcome variables. A total of 88% patients achieved a reduction in the final HAMD-D score at the end of treatment. The total HAM-D score reduced to 10 or less in 73% patients at 6 weeks and by this week 76% and 92% patients achieved a score of 1 or 2 for CGI-S and CGI-I, respectively. The mean Clinical Anxiety Scale score reduced from 12.6 at the baseline to 4.4 at the end of treatment. Safety data was evaluated in all 112 patients and paroxetine was well tolerated. Adverse events were experienced by 10% of patients of whom 4% were dropped from the study. Nausea was the commonest adverse event reported. CONCLUSION: The study shows that paroxetine is an effective, well-tolerated, and safe drug for the treatment of depression. PMID- 12585375 TI - View box case-11. Synovial osteochondromatosis with osteoarthrosis. PMID- 12585374 TI - Radiologic features of Hallervorden Spatz Disease. PMID- 12585376 TI - Obesity: a rationale approach for a family physician. PMID- 12585377 TI - Evolving role of biliary stenting in iatrogenic jaundice. PMID- 12585378 TI - Catalytic enantioselective allylation with chiral Lewis bases. AB - The advent of chiral Lewis base-promoted allylation of aldehydes has opened a new direction in the catalytic enantioselective construction of homoallylic alcohols. This short review outlines the conceptual framework for the creation of this new process and the interplay of mechanistic investigations and synthetic studies that have conspired to produce a useful new reaction. The current state-of-the art in catalyst design and application of the reaction in synthesis are briefly illustrated. PMID- 12585379 TI - Imprinted polymers-tailor-made mimics of antibodies and receptors. AB - The technique of molecular imprinting allows the formation of specific recognition sites in synthetic polymers through the use of templates or imprint molecules. These recognition sites mimic the binding sites of antibodies and other biological receptor molecules. Molecularly imprinted polymers can therefore be used in applications relying on specific molecular binding events. The stability, ease of preparation and low cost of these materials make them particularly attractive. This review focuses on recent developments and advances in the field of molecularly imprinted materials, with special emphasis on applications in immunoassays and sensors recently developed by our group and by others. PMID- 12585381 TI - Selective cross-catenation of Pd(II) and Pt(II) coordination rings. AB - The reversible cross-catenation of two different coordination rings (Pd(II)- and Pt(II)-linked rings) has been achieved by using the labile nature of the Pd-N interaction and efficient hydrophobic contact between the rings. PMID- 12585380 TI - Peptide-polymer bioconjugates: hybrid block copolymers generated via living radical polymerizations from resin-supported peptides. AB - A novel strategy for the preparation of peptidic-synthetic bioconjugate block copolymers is based upon sequential condensation and living radical addition polymerizations, each performed upon a solid support. PMID- 12585382 TI - A methylene-bis-triazolium ligand precursor in an unusual rearrangement of norbornadiene to nortricyclyl. AB - Reaction of [(nbd)RhCl]2 with a chelating bis-[1,2,4]-triazolium salt gives a nortricyclyl Rh complex. PMID- 12585383 TI - Crystal engineering of the composition of pharmaceutical phases. AB - The carboxylic acid-pyridine supramolecular heterosynthon can be exploited to predictably generate binary crystalline phases involving rac-ibuprofen, rac flurbiprofen or aspirin. PMID- 12585384 TI - Photochemical and thermal ligand exchange in a ruthenium(II) complex based on a scorpionate terpyridine ligand. AB - A ruthenium(II) complex containing a 1,10-phenanthroline unit and a terpyridine fragment covalently linked to a benzonitrile group has been synthesised; coordination and decoordination of the benzonitrile group can be induced thermally and photochemically respectively, in an acetone-water mixture. PMID- 12585385 TI - Ring opening metathesis polymerization on non-covalently functionalized single walled carbon nanotubes. AB - Norbornene polymerization has been initiated selectively on the surface of single walled carbon nanotubes (SWNTs) via a specifically adsorbed pyrene-linked ROMP initiator, resulting in a homogeneous non-covalent poly(norbornene) coating. PMID- 12585386 TI - Polymer-supported formamides as reusable organocatalysts for allylation of aldehydes with allyltrichlorosilane. AB - New types of polymer-supported formamides have been synthesized from chloromethylated resins and formamides; it was found that the polymers worked well as organocatalysts in the allylation of aldehydes with allyltrichlorosilane to afford homoallylic alcohols in high yields; the polymers were easily recovered and reused several times without loss of activity. PMID- 12585387 TI - An anionic organic mixed-valence system with a remarkably well-resolved vibrational structure in its intervalence band. AB - The reduction of a phenylene-bridged bis(dioxaborine) affords a strongly delocalised organic mixed-valence system; for the first time details of the vibrations coupled to the electron transfer have been extracted from the intervalence band. PMID- 12585388 TI - Studies towards the total synthesis of an epoxy isoprostane phospholipid, a potent activator of endothelial cells. AB - We report studies toward the total synthesis of an epoxy isoprostane, namely the preparation of compound 9 which is an analogue of the elimination product 7 of the naturally occurring epoxy isoprostane 4 by a straightforward route using a three-component coupling, and have shown by several spectroscopic criteria that it closely resembles the natural material. PMID- 12585389 TI - Further reactions of phenyldimethylsilyllithium with N,N-dimethylamides. AB - Phenyldimethylsilyllithium reacts with several N,N-dimethylamides, and the intermediates, formulated here as successively a carbene and an alpha silyllithium species, may be trapped with nucleophiles and electrophiles, respectively, although not always with the nucleophile or electrophile of your choice. PMID- 12585390 TI - Reactions of phenyldimethylsilyllithium with beta-N,N-dimethylaminoenones. AB - Phenyldimethylsilyllithium reacts with the beta-N,N-dimethylaminoenones 1 and 7, with the enal 5, and with ethyl beta-N,N-dimethylaminoacrylate 9 to give the corresponding alpha,beta-unsaturated carbonyl compounds with a beta phenyldimethylsilyl group, but in the last case only when the reaction mixture is given a mysteriously brief treatment with methyl iodide before workup. PMID- 12585391 TI - The reaction of phenyldimethylsilyllithium with N-phenylpyrrolidone. AB - Phenyldimethylsilyllithium reacts with N-phenylpyrrolidone 5 to give the known tetracyclic amines [2,3,3a,3b,4,5,6,11b-octahydro-3aalpha,3balpha, 11balpha-1 phenyl-1H-dipyrrolo(1,2a:3',2'c)quinoline and its 3bbeta isomer] 6 and 7. PMID- 12585392 TI - A practical access to acyl radicals from acyl hydrazides. AB - Various acyl radicals can be generated from the corresponding acyl triphenylmethyldiazo derivatives, produced by in situ oxidation of hydrazide precursors with phenylseleninic acid. PMID- 12585393 TI - Electrochemical synthesis of novel polypyrrole microstructures. AB - Polypyrrole microstructures with unusual morphologies have been synthesized by direct electrochemical oxidation of pyrrole in beta-naphthalenesulfonic acid aqueous solution. PMID- 12585394 TI - A novel nanostructure of nickel nanotubes encapsulated in carbon nanotubes. AB - A novel nanostructure of Ni nanotubes encapsulated in carbon nanotubes has been obtained via the pyrolysis of C2H2 on an array of Ni nanotubes in an alumina membrane support and a possible mechanism has been proposed. PMID- 12585395 TI - A new thioether functionalized organic-inorganic mesoporous composite as a highly selective and capacious Hg2+ adsorbent. AB - A new thioether functionalized organic-inorganic ordered mesoporous composite as a highly selective and capacious Hg2+ adsorbent was synthesized by one-step co condensation of (1,4)-bis(triethoxysilyl)propane tetrasufide (BTESPTS, (CH3CH2O)3Si(CH2)3S-S-S-S(CH2)3Si(OCH2CH3)3) and tetraethoxysilane (TEOS), with tri-block copolymer poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) EO20PO70EO20 as template. PMID- 12585396 TI - Development of silver film via thermal decomposition of layered silver alkanecarboxylates for surface-enhanced Raman spectroscopy. AB - This work demonstrates that optically tunable, SERS-active Ag films can be reproducibly fabricated on glass by the thermal decomposition of layered silver alkanecarboxylates. PMID- 12585397 TI - Mechanistic studies on ethylene silylation with chlorosilanes catalysed by ruthenium complexes. AB - Silylation of ethylene by chlorosilanes is catalysed by ruthenium complexes. Mechanistic investigations reveal the presence of a complicated network of reactions leading to new sigma-silane, ethylene and silyl complexes. PMID- 12585398 TI - Cationic arylation through photo(sensitised) decomposition of diazonium salts. Chemoselectivity of triplet phenyl cations. AB - The photodediazoniation of some 4-X-phenyldiazonium tetrafluoborates in MeCN leads to the singlet phenyl cations (X = H, tert-butyl, NMe2, CN), which add to the solvent yielding the corresponding acetanilides. Triplet sensitisation, however, leads to the triplet phenyl cation, which is reduced in neat solvent and is trapped by pi nucleophiles (allyltrimethylsilane and benzene), resulting in an ionic analogue of the Meerwein or Gomberg arylations. With the 4-nitro derivative intersystem crossing prevails over dediazoniation from the singlet and with the 4 cyano competes with it, so that in those cases the triplet phenyl cation is formed also upon direct irradiation. PMID- 12585399 TI - Knoevenagel condensation reaction in a membrane microreactor. AB - A multi-channel membrane microreactor was fabricated and tested for Knoevenagel condensation of benzaldehyde and ethyl cyanoacetate using Cs-exchanged faujasite NaX as the catalyst; the membrane microreactor achieves supra-eqiulibrium conversion at higher product purity. PMID- 12585400 TI - Effects of a nonplanar porphyrin rings on the spin-spin interactions in low-spin ferric porphyrin radical cationst. AB - Low-spin ferric porphyrin radical cations formed by the oxidation of chloro(meso tetraalkylporphyrinato)iron(III) followed by the addition of bulky 2 methylimidazole show antiferromagnetic coupling, which is interpreted in terms of the interaction between porphyrin a2u and iron d(xy), orbitals caused by the S4 ruffling of the porphyrin core. PMID- 12585401 TI - Naphthalene in the higher triplet excited state. AB - Naphthalene in the higher triplet excited state Np(Tn) was generated from the two step excitation method using two-colour two-laser flash photolysis technique and the lifetime of Np(Tn) was estimated to be 4.5 ps from the triplet energy quenching by quenchers such as p-dichlorobenzene, o-dicyanobenzene and carbon tetrachloride. PMID- 12585402 TI - Heat inhibited reactions. AB - Acyl transfer from p-nitrophenyl trimethylacetate to hydrogen peroxide in millimolar aqueous solutions of an amphiphilic poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) triblock copolymer slows down as the temperature is raised due to partitioning of the hydrophobic ester into heat induced micelles. PMID- 12585403 TI - Synthesis and photophysics of a linear non-covalently linked porphyrin-fullerene dyad. AB - The synthesis and characterization of a new pyridinofullerene ligand capable of forming axially symmetric complexes with ZnTPP is reported; molecular modelling studies, 1H NMR, UV-Vis spectroscopy and fluorescence quenching data support formation of a strong complex between the new ligand and ZnTPP. PMID- 12585404 TI - The synthesis of a single enantiomer of a major alpha-mycolic acid of Mycobacterium tuberculosis. AB - We report a synthesis of a single enantiomer of a dicyclopropane containing mycolic acid from Mycobacterium tuberculosis; this method can be simply varied to modify the chain lengths or the absolute stereochemistry of either cyclopropane. PMID- 12585405 TI - Increasing the crystallisation temperature to access new spin clusters: conversion of [Ni8(cit)6(OH)2(H2O)2]10- to [Ni8(cit)6(OH)2]10-. AB - The role of temperature in the formation of high nuclearity nickel(II) citrate spin clusters is explored, revealing how changes in structure and hence magnetic properties can be triggered through desolvation and ligand reorganisation. PMID- 12585406 TI - Effect of pressure and temperature upon tin alkoxide-promoted ring-opening polymerisation of epsilon-caprolactone in supercritical carbon dioxide. AB - Ring-opening polymerisation of epsilon-caprolactone in supercritical carbon dioxide is slowed down by a carbonation reaction, resulting in a positive volume of activation and a higher energy of activation as compared to polymerisation in a regular hydrocarbon solvent. PMID- 12585407 TI - Pd.Et3B-catalyzed alkylation of amines with allylic alcohols. AB - A combination of catalytic amounts of Pd (0.05 mmol) and Et3B (0.3 mmol) promotes allylic alkylation of primary and secondary aromatic and aliphatic amines (1.0 mmol) by the direct use of allylic alcohols, providing tertiary amines in excellent yields under mild conditions (room temperature approximately 50 degrees C). PMID- 12585408 TI - Ordered mesoporous phosphosilicate glass electrolyte film with low area specific resistivity. AB - Ordered mesoporous phosphosilicate glass electrolyte film exhibiting low area specific resistivity (ASR) compared with those of Nafion has been synthesized using non-ionic surfactant as the structure-directing agent. PMID- 12585410 TI - Computer screen as a programmable light source for visible absorption characterization of (bio)chemical assays. AB - Visible absorption features suitable for color recognition and micro-plate reading of a standard bioassay are performed by the combination of a computer screen used as a programmable light source and a web camera as detector. The method provides in this way a highly available platform for 'home tests' or 'self tests', where the requirement is to monitor well defined assays and the use of economical instrumentation is advantageous. PMID- 12585409 TI - Controlled formation of biosilica structures in vitro. AB - Herein we describe the controlled formation of biosilica structures by manipulation of the physical reaction environment; we were able to synthesize arched and elongated silica structures using a synthetic peptide; the results presented here are evidence that in vitro biocatalysis may be controlled in order to form desired silica structures. PMID- 12585411 TI - Preparation of a mesoporous silicon boron nitride via a non-aqueous sol-gel route. AB - A silicon boron imide based gel Si3B(NH)x(NH2)y(NMe2)z was prepared via catalytic ammonolysis of tris[tris(dimethylamino)silylamino]boron B[HNSi[N(CH3)2]3]3; pyrolysis of the gel under ammonia at 1,000 degrees C gives a mesoporous silicon boron nitride with high surface area and narrow pore size distribution. PMID- 12585412 TI - Correlation between structure, aggregation behaviour and cellular toxicity of anti-HIV catanionic analogues of galactosylceramide. AB - The self-association process of catanionic analogues of galactosylceramide and in particular the arrangement of their hydrophobic part seems to play a key role in their cellular toxicity PMID- 12585413 TI - Sulfonic acid-functionalized silica through quantitative oxidation of thiol groups. AB - A simple procedure for the preparation of amorphous silica containing thiol groups which quantitatively affords sulfonic acid groups has been developed, resulting in site densities and activity for the esterification of acetic acid with methanol greater than a commercial Nafion silica composite. PMID- 12585414 TI - A novel synthesis of metallogermacyclopropane and molybdenum bis(iminophosphorano)carbene complexes from bisgermavinylidene. AB - The reaction of bisgermavinylidene [(Me3SiN=PPh2)2C=Ge-->Ge=C(PPh2=NSiMe3)2] (1) with M(CO)5(THF) ( M = Cr, W, Mo) afforded the metallagermacyclopropane [(Me 3SiN=PPh2)2CGeM(CO)3[M(CO)5]] [M = W (2), Cr (3), Mo (4)]; in one of the reactions, compound 4 reacts further to give a "pincer" carbene complex [(CO)3Mo[C(Ph2P=NSi Me3)2]] (5); the X-ray structures of compounds 2 and 5 have been determined. PMID- 12585415 TI - In situ co-crystallisation as a tool for low-temperature crystal engineering. AB - Synthesis and single-crystal X-ray diffraction of a series of low-melting co crystals of pyrazine and n-alkyl carboxylic acids demonstrates in situ co crystallisation as a versatile tool for low-temperature crystal engineering. PMID- 12585416 TI - Novel polyene dyes for highly efficient dye-sensitized solar cells. AB - We have developed an efficient and novel polyene-dye-sensitized nanocrystalline TiO2 solar cells producing a 6.8% solar energy-to-electricity conversion efficiency (eta) under AM 1.5 irradiation (100 mW cm(-2)): short-circuit current density (Jsc), 12.9 mA cm(-2), open-circuit photovoltage (Voc), 0.71 V, fill factor (ff), 0.74. PMID- 12585417 TI - Novel thymidine-based organogelators and their gelation behaviours. AB - We report on a synthesis of novel thymidine based organogelators and a study of their gelation types in relation to structure and solvent, using various data acquired through FT-IR, SEM images and differential scanning calorimetry. PMID- 12585418 TI - Direct synthesis of aligned silicon carbide nanowires from the silicon substrates. AB - Aligned silicon carbide nanowires were synthesized directly from the silicon substrates via a novel catalytic reaction with a methane-hydrogen mixture at 1,100 degrees C, with a mean diameter of 40 nm and length of 500 microm; they consist of a single-crystalline zinc blende structure crystal in the [111] growth direction; X-ray diffraction, Raman, and infrared spectroscopy confirm the synthesis of high-purity silicon carbide nanowires. PMID- 12585419 TI - Gas phase transport of gold with gold(III) oxide and carbon monoxide. AB - A transport reaction of gold occurs during the reaction of Au2O3 with CO at temperatures around 400 degrees C. The reaction proceeds through a short-lived gas phase species of Au; analysis of quartz powder substrates placed downstream of the AU2O3 indicates volatilisation yields of a few wt% AU2O3. PMID- 12585420 TI - A novel stereoselective one-pot conversion of alcohols into alkyl halides mediated by N,N'-diisopropylcarbodiimide. AB - Alcohols can be converted in high yields to the corresponding alkyl halides in a one-pot procedure via the corresponding O-alkylisourea; very short reaction times are possible when microwave irradiation is used. PMID- 12585421 TI - In situ cyclodextrin-based homogeneous incorporation of metal (M = Pd, Pt, Ru) nanoparticles into silica with bimodal pore structure. AB - In situ cyclodextrin (CD)-based homogeneous incorporation of metal (M = Pd, Pt, Ru) nanoparticles into silica with bimodal pore structure has been realized by utilizing the self-assembly aggregation and inclusion capability of the CDs. PMID- 12585422 TI - Synthesis, catalytic properties and biological activity of new water soluble ruthenium cyclopentadienyl PTA complexes [(C5R5)RuCl(PTA)2] (R = H, Me; PTA = 1,3,5-triaza-7-phosphaadamantane). AB - The new water soluble ruthenium complexes [(C5R5)RuCl(PTA)2] (R = H, Me; PTA = 1,3,5-triaza-7-phosphaadamantane) were synthesised and characterised. Their evaluation as regioselective catalysts for hydrogenation of unsaturated ketones in aqueous biphasic conditions and as cytotoxic agents towards the TS/A adenocarcinoma cell line is briefly presented. PMID- 12585423 TI - Rapid reduction of pentavalent antimony by trypanothione: potential relevance to antimonial activation. AB - The dithiol trypanothione can reduce an antiparasitic pentavalent antimony agent to trivalent rapidly; this reduction process is both pH and temperature dependent and trypanothione may therefore play an important role in the activation of the drug. PMID- 12585424 TI - Anomalous adsorption of nitrogen and argon in silver exchanged zeolite A. AB - A new adsorbent based on zeolite AgA having N2 adsorption capacity of 22.3 cc g( 1) at 101.3 kPa and N2/O2 selectivity in the range of 5 to 14.6 at 303 K, the highest known so far for any zeolite A type of adsorbent, is reported; furthermore, this adsorbent also shows argon selectivity over oxygen. PMID- 12585425 TI - Fast peroxyoxalate chemiluminescence for minimized analytical separation systems. AB - The maximum intensity, Imax, and time required to reach the maximum emission, taumax, for 1-aminopyrene monitored in 1,1'-oxalyldi-4-methylimidazole (OD4MI) chemiluminescence (CL) reactions are approximately 61 times higher and 16 times faster than their respective values for bis(2,4,6-trichlorophenyl)oxalate (TCPO) CL reactions in the presence of imidazole (ImH). PMID- 12585426 TI - Unexpected isolation, and structural characterization, of a beta-hydrogen containing sigma-alkylpalladium halide complex in the course of an intramolecular heck reaction. synthesis of polycyclic isoquinoline derivatives. AB - The isolation of a stable beta-hydrogen-containing R-PdLn-X complex (R = alkyl; X = halide) issued from a Heck reaction is reported together with some aspects of its reactivity. PMID- 12585427 TI - Synthesis of a stibindolyl anion from trimesitylantimony and potassium. AB - 2-(3',5'-Dimethylphenyl)-5,7-dimethylstibindolyl potassium-pmdeta (1) (pmdeta = pentamethyldiethylenetriamine) is obtained from trimesitylantimony, potassium and the amine ligand in tetrahydrofuran; crystals of 1 consist of stacks of (pmdeta)K+ ions and stibindolyl anions with alternating Sb-K and eta5-SbC4-K interactions. PMID- 12585428 TI - A novel diastereoselective route to alpha-hydroxyalkyl dihydropyrans using a hetero Diels-Alder/allylboration sequence. AB - The development of a new strategy for the synthesis of alpha-hydroxyalkyl dihydropyrans is reported. This approach is based on a tandem hetero[4+2]/allylboration reaction. PMID- 12585429 TI - Factors controlling the relative stabilities of cis- and trans-[PtX2L2] isomers: Chatt and Wilkins--50 years on. AB - DFT computations reveal that cis-trans preferences in [PtX2(PR3)2] species are due to a combination of electrostatics, pi-backbonding, antisymbiosis, and solvation effects. PMID- 12585430 TI - A divergent approach to the precise synthesis of giant organometallic dendrimers using platinum-acetylides as building blocks. AB - Giant platinum-acetylides dendrimers were precisely synthesized by a divergent method; the sixth generation dendrimer, the diameter of which is larger than 10 nm, has 189 Pt atoms per molecule, and its molecular weight is as high as 139750. PMID- 12585431 TI - Synthesis of tripodal [2]rotaxanes: high concentration principlet. AB - Symmetrical and non-symmetrical tripodal [2]rotaxanes (1) incorporating 1,1' disubstituted-4,4'-bipyridinium cations (2) and 34-crown-10 (3) have been prepared directly from 4,4'-bipyridine or from monocationic intermediates in high yields at room temperature and atmospheric pressure under conditions that permit the use of high reagent concentrations (0.1-0.2 M, 150-200 g(-1)). PMID- 12585432 TI - Alignment control of a cyanine dye using a mesoporous silica film with uniaxially aligned mesochannels. AB - The feasibility of alignment control of low-molecular-weight guest species by controlling the orientation of mesochannels in a mesoporous silica host has been shown for the first time; spectroscopically anisotropic absorption behaviour of a cyanine dye was observed upon its incorporation into the mesochannels of a mesoporous silica film with uniaxially aligned porous structure, proving that the preferred alignment of the dye molecules was achieved on a macroscopic scale. PMID- 12585433 TI - An artificial antenna complex containing four Ru(bpy)3(2+)-type chromophores as light-harvesting components and a Ru(bpy)(CN)4(2-) subunit as the energy trap. A structural motif which resembles the natural photosynthetic systems. AB - A cyclam subunit bearing four light-harvesting Ru(bpy)3(2+)-type species is connected to a Ru(II) compound having low-lying excited states (energy trap) by hydrogen bonding: the light absorbed by the light-harvesting elements is funnelled to the energy trap with high efficiency. PMID- 12585434 TI - One-pot synthesis of 2,5-diethynyl-3,4-dibutylthiophene substituted multitopic bipyridine ligands: redox and photophysical properties of their ruthenium(II) complexes. AB - A facile and original synthesis of four 2,2'-bipyridine (bipy) ligands grafted with thiophene subunits is described using phase transfer experimental conditions: related Ru(II) complexes exhibit well-defined redox and photophysical properties which were probed by cyclic voltammetry, UV-vis, steady-state emission and transient absorption spectroscopy. PMID- 12585435 TI - Unique crystal structures of donor-acceptor complexes: crossed arrangement of two charge-transfer columns. AB - Unique crystal structures containing two kinds of crossed charge-transfer (CT) columns are demonstrated by cocrystals consisting of ion pairs of acceptors of methyl viologen and anthraquinone disulfonate derivatives and a donor, hydroquinone, mediated by electrostatic and donor-acceptor (DA) interactions. PMID- 12585436 TI - Resident recruitment. AB - This article has introduced the reader to the critical components of successful recruitment of radiology residents. With particular attention to the ACGME institutional and program requirements regarding resident recruitment, and an explanation of the support systems (ERAS and NRMP) currently available to those involved in applicant review and selection, the article has sought to delineate a sensible approach to recruitment. Successful recruiters have mastered the essentials of these programs and have learned to adapt the programs to their needs. As new program directors work with their departments' resident selection committees, they will identify the factors that faculty and current residents cite as most important in the successful selection of new residents. By structuring the application review process, exploiting the power of the ERAS, and crafting a purposeful and friendly interview process, radiology residency directors can find and recruit the residents who best match their programs. PMID- 12585437 TI - The ACGME and the residency review committee: external program review. Accreditation Council for Graduate Medical Education. PMID- 12585438 TI - Responsibilities of the program director. PMID- 12585439 TI - Program documents: policies and guidelines. PMID- 12585440 TI - Resident research. PMID- 12585441 TI - Institutional requirements. AB - The residency review committee's program requirements guide the development of the details of each ACGME-accredited residency program. The sponsoring institution, however, has the overall responsibility, through the institutional requirements, to ensure that all facets of high-quality graduate medical education are available for all residents in these programs. It is imperative that there be close interaction between residency program directors, departmental education coordinators, and the GME officials of the institution, especially as ACGME regulations have become more complex. PMID- 12585442 TI - Evaluation of residents, faculty, and program. PMID- 12585443 TI - Internal program review. PMID- 12585444 TI - The role of the residency coordinator. PMID- 12585445 TI - Chief residency. PMID- 12585446 TI - Radiologic pathology correlation course at the Armed Forces Institute of Pathology. PMID- 12585447 TI - American Board of Radiology. PMID- 12585448 TI - Program directors as conflict managers. PMID- 12585449 TI - Funding of graduate medical education. PMID- 12585450 TI - Internet resources for education in radiology. PMID- 12585451 TI - Organized radiology. PMID- 12585452 TI - Educational infrastructure for radiology residency programs. PMID- 12585453 TI - Teaching radiology residents, and radiology residents as teachers. PMID- 12585454 TI - Quantitative synthetic polymer mass spectrometry workshop at the National Institute of Standards and Technology. PMID- 12585456 TI - Shrinking the LC landscape. PMID- 12585455 TI - Measuring DNA synthesis rates with stable isotopes. PMID- 12585457 TI - Finding a real gem. PMID- 12585458 TI - Raman on the run. PMID- 12585459 TI - Experimental studies of electroosmotic flow dynamics in microfabricated devices during current monitoring experiments. AB - Electroosmotic flow (EOF) was monitored in glass microfluidic devices at rates up to 2 Hz with a precision of 0.2-1.0% using a technique based on the periodic photobleaching of a dilute, neutral fluorophore added to the running buffer. This EOF monitoring method was used to examine the performance of the current monitoring technique for measuring an average electroosmotic flow in a microfluidic device with a cross-T design. Flow measurements made with the current monitoring method gave a precision of 0.4-2.2%, but the periodic photobleaching method shows that the current monitoring technique causes changes in EOF as high as 41% during a single experiment. The periodic photobleaching method for EOF monitoring was also used to study EOF in channels on opposite sides of a cross-channel intersection. The opposite channels were shown to exhibit substantially different EOF dynamics during a current monitoring experiment as well as different steady-state EOF rates during normal operating conditions. PMID- 12585460 TI - Universal two-dimensional HPLC technique for the chemical analysis of complex surfactant mixtures. AB - Two-dimensional HPLC was applied for the first time to the analysis of complex surfactant mixtures. In the first dimension, ion chromatographic-type separations were performed on a diol column eluted by an acetonitrile-water (0.1% trifluoroacetic acid) gradient. Using this new technique, cationic and amphoteric surfactants were not retained at all, nonionic surfactants exhibited a weak and essentially unspecific retention, and anionic surfactants were retained mainly according to their functional group. Rather than detecting the analytes immediately after this first separation, successive fractions were automatically and quantitatively transferred to parallel C2 (dimethyl) and C4 (butyl) reversed phase columns using an innovative setup. The second dimension of the separation then took place, by which the analytes were separated according to their hydrophobicity. Surfactants from all four classes, cationic, amphoteric, nonionic, and anionic, were separated simultaneously in single 54-min two dimensional HPLC runs. The suitability of the method for quantitative measurements was demonstrated. PMID- 12585461 TI - Polysodium N-undecanoyl-L-leucylvalinate: a versatile chiral selector for micellar electrokinetic chromatography. AB - Dipeptide micelle polymers are a new class of polymeric surfactants of which the polysodium undecanoyl-L-leucylvalinate (poly-L-SULV) was found to be a broadly applicable chiral selector for micellar electrokinetic chromatography. This negatively charged dipeptide micelle polymer is a high molecular weight compound with large countercurrent mobility, zero critical micelle concentration, low aggregation number, and high solubility in water or water-organic solvents. In an extensive chiral screening program, enantioseparation of 75 racemic compounds was tested with poly-L-SULV as chiral pseudostationary phase in neutral pH and basic pH background electrolytes. A total of 58 out of 75 racemic compounds could be resolved after choosing an appropriate concentration of poly-L-SULV. Although anionic chiral analytes are difficult to resolve using poly-L-SULV, the percent success rate for chiral resolution of cationic (77%) and neutral (85%) racemates was very high. Aspects regarding electrostatic, steric, hydrophobic, and hydrogen bonding interactions of this dipeptide micelle polymer with various classes of chiral analytes are discussed. PMID- 12585463 TI - Light scattering and light absorbance separated by extended multiplicative signal correction. application to near-infrared transmission analysis of powder mixtures. AB - The extended multiplicative signal correction (EMSC) preprocessing method allows a separation of physical light-scattering effects from chemical (vibrational) light absorbance effects in spectra from, for example, powders or turbid solutions. It is here applied to diffuse near infrared transmission (NIT) spectra of mixtures of wheat gluten (protein) and starch (carbohydrate) powders, linearized by conventional log(1/T). Without any correction for uncontrolled light scattering variation between the powder samples, these absorbance spectra could give reasonable predictions of the analyte (gluten), but only when using multivariate calibration with a much more complex model than expected. Standard MSC preprocessing did not work for these data at all; it removed too much analyte information. However, the EMSC preprocessing yielded powder spectra that obeyed Beer's Law more or less as if they had been obtained from transparent liquid solutions, apparently by isolating the chemical light absorption from additive, multiplicative, and wavelength-dependent effects of uncontrolled light-scattering variations. The model-based EMSC and its converse, the extended inverted signal correction (EISC), gave rather complete descriptions of the diffuse absorbance spectra and virtually indistinguishable performance in the calibration set and the test set of samples. PMID- 12585462 TI - Creation of an on-chip enzyme reactor by encapsulating trypsin in sol-gel on a plastic microchip. AB - Trypsin-encapsulated sol-gel was fabricated in situ onto a plastic microchip to form an on-chip bioreactor that integrates tryptic digestion, separation, and detection. Trypsin-encapsulated sol-gel, which is derived from alkoxysilane, was fabricated within a sample reservoir (SR) of the chip. Fluorescently labeled ArgOEt and bradykinin were digested within the SR followed by electrophoretic separation on the same chip. The plastic microchip, which is made from poly(methyl methacrylate), generated enough electroosmotic flow that substrates and products could be satisfactorily separated. The sol-gel in the SR did not alter the separation efficiency of each peak. With the present device, the analytical time was significantly shortened compared to conventional tryptic reaction schemes. This on-chip microreactor was applicable to the digestion of protein with multiple cleavage sites and separation of digest fragments. Furthermore, the encapsulated trypsin exhibits increased stability, even after continuous use, compared with that in free solution. PMID- 12585464 TI - Synergistic pseudo-hydroxide extraction: synergism and anion selectivity in sodium extraction using a crown ether and a series of weak lipophilic acids. AB - The nature of the weak lipophilic acid used in synergistic combination with a model crown ether cation host was shown to have a strong effect on the strength and selectivity of sodium hydroxide separation from alkaline aqueous salt solutions. Sodium ion-pair extraction employing only cis-syn-cis-dicyclohexano-18 crown-6 (1) in nitrobenzene (NB) was correlated with the standard Gibbs energy (deltaG(p)o) of anion partitioning into NB and was notably weak and nonselective for the hydroxide ion, in accord with Hofmeister bias. The Hofmeister order can be selectively overcome for NaOH by utilization of acid-base chemistry coupled with complexation of sodium ion in the NB phase. Upon addition of a lipophilic organic acid into the solution of 1 in NB, sodium extraction was selectively enhanced due to the initiation of an exchange reaction between the aqueous sodium ion and the ionizable proton of the organic acid. A series of weak lipophilic hydroxy acids (HA) including fluorinated alcohols and phenols was tested. The resulting synergistic pseudo-hydroxide extraction correlates with the pKa of the employed HA; the most acidic cation exchangers provide the greatest synergism. The synergistic factor obtained using a fluorinated benzyl alcohol 7 was as high as 256. Ion-pair extraction of neutral sodium salts was not changed or only mildly enhanced by addition of HA into the NB solution of 1. This enhancement was explained by hydrogen bonding of HA with the anion as related to the hardness of the anion and the acidity of HA. In comparison with the synergism observed for NaOH, this enhancement was weak and unable to overcome the Hofmeister effect. Examination of extraction selectivity revealed that the combination of 1 and 7 preferentially extracted NaOH over all other sodium salts, including the normally preferred nitrate and perchlorate salts. Quantitative recovery of NaOH from the NB phase was demonstrated via hydrolysis of the organic acid upon a single contact of the loaded solvent with water. PMID- 12585465 TI - Selective determination of beryllium(II) ion at picomole per decimeter cubed levels by kinetic differentiation mode reversed-phase high-performance liquid chromatography with fluorometric detection using 2-(2'-hydroxyphenyl)-10 hydroxybenzo[H]quinoline as precolumn chelating reagent. AB - A highly sensitive and selective method for the determination of the Be(II) ion has been developed by the use of reversed-phase high-performance liquid chromatography (HPLC) with fluorometric detection using 2-(2'-hydroxyphenyl)-10 hydroxybenzo[h]quinoline (HPHBQ) as a precolumn (off-line) chelating reagent. The reagent HPHBQ has been designed to form the kinetically inert Be chelate compatible with high fluorescence yield, which is appropriate to the HPLC fluorometric detection system. The Be-HPHBQ chelate is efficiently separated on a LiChrospher 100 RP-18(e) column with a methanol (58.3 wt %)-water eluent containing 20 mmol kg(-1) of tartaric acid and is fluorometrically detected at 520 nm with the excitation at 420 nm. Under the conditions used, the concentration range of 20-8,000 pmol dm(-3) of Be(II) ion can be determined without interferences from 10 micromol dm(-3) each of common metal ions, typically Al(III), Cu(II), Fe(III), and Zn(II), and still more coexistence of Ca(II) and Mg(II) ions at 0.50 mmol dm(-3) and 5.0 mmol dm(-3), respectively, is tolerated. The detection limit (3a baseline fluctuation) is 4.3 pmol dm(-3) (39 fg cm(-3)). The extraordinarily high sensitivity with toughness toward the matrix influence was demonstrated with the successful application to environmental Be analyses, such as determination of Be in rainwater and tap water. PMID- 12585466 TI - Collision-activated infrared multiphoton dissociation in a quadrupole ion trap mass spectrometer. AB - We propose and demonstrate a new method for multiple-stage mass spectrometry (MSn), collision-activated infrared multiphoton dissociation (CA-IRMPD), which is very effective for the quadrupole ion trap mass spectrometer (QITMS). CA-IRMPD uses a combination of focused laser irradiation (beam radius, approximately 0.4 mm) and collisional activation by a supplemental AC voltage between endcap electrodes. This combination enables IRMPD, which has conventionaLly been ineffective above 10(-4) Torr, to be used under a standard bath gas pressure of 2 8 mTorr. CA-IRMPD can produce richer spectra of product ions than CID or IRMPD while maintaining high sensitivity and mass resolution; thus, it will contribute to an accurate determination of peptide sequences. PMID- 12585467 TI - The importance of chromatographic separation in LC/MS/MS quantitation of drugs in biological fluids: detection, characterization, and synthesis of a previously unknown low-level nitrone metabolite of a substance P antagonist. AB - The observation of an interference peak in plasma samples from dogs dosed with compound I led to the discovery of an unidentified metabolite. The unknown metabolite had the same molecular weight as the parent drug, and their fragmentation profiles were also quite similar. LC/MS/ MS analysis of the plasma extracts of dogs and rats dosed with I and its deuterium-labeled analogue suggested a nitrone structure for the unknown metabolite. Synthesized nitrone matched the unknown metabolite with identical retention time and nearly identical fragmentation profile. The nitrone slowly decomposed in acidic aqueous solution at ambient temperature and also underwent in-source, thermal-induced hydrolysis during electrospray ionization mass spectrometric analysis. The reaction of the nitrone with diethyl acetylenedicarboxylate readily generated a [2 + 3] cycloaddition product. The example shown here clearly demonstrates that precautions must be taken when LC/MS/MS quantitation is conducted in the selected reaction monitoring mode. PMID- 12585468 TI - Intensity-based statistical scorer for tandem mass spectrometry. AB - We describe a new statistical scorer for tandem mass spectrometry. The scorer is based on the probability that fragments with given chemical properties create measured intensity levels in the experimental spectrum. The scorer's parameters are computed using a fully automated procedure. Benchmarking the new scorer on a large set of experimental spectra, we show that it performs significantly better than the widely used cross-correlation scoring algorithm of Eng et al. (Eng, J. K; McKormack, A. L.; Yates, J. R. J. Am. Soc. Mass Spectrom. 1994, 5, 976-989.). PMID- 12585469 TI - Absolute quantification of the G protein-coupled receptor rhodopsin by LC/MS/MS using proteolysis product peptides and synthetic peptide standards. AB - Methods for the absolute quantification of a membrane protein are described using isotopically labeled or unlabeled synthetic peptides as standards. Synthetic peptides are designed to mimic peptides that are cleaved from target analyte proteins by proteolytic or chemical digestion, and the peptides selected serve as standards for quantification by LC/MS/MS on a triple quadrupole mass spectrometer. The technique is complementary to relative quantification techniques in widespread use by providing absolute quantitation of selected targets with greater sensitivity, dynamic range, and precision. Proteins that are found to be of interest by global proteome searches can be selected as targets for quantitation by the present method. This method has a much shorter analytical cycle time (minutes versus hours for the global proteome experiments), making it well suited for high-throughput environments. The present approach using synthetic peptides as standards, in conjunction with proteolytic or chemical cleavage of target proteins, allows mass spectrometry to be used as a highly selective detector for providing absolute quantification of proteins for which no standards are available. We demonstrate that quantification is simple and reliable for the integral membrane protein rhodopsin with reasonable recoveries for replicate experiments using low-micromolar solutions of rhodopsin from rod outer segments. PMID- 12585470 TI - Off-line solid-phase microextraction and capillary electrophoresis mass spectrometry to determine acidic pesticides in fruits. AB - A method based on solid-phase microextraction (SPME) and capillary electrophoresis/mass spectrometry (CE/ MS) is described for determining simultaneously five acidic pesticides (o-phenylphenol, ioxynil, haloxyfop, acifluorfen, picloram) in fruits. The CE device is coupled to an electrospray interface by a commercial sheath-flow adapter. Emphasis is placed on fulfillment of the speed and sensitivity requirements. The best separation is achieved using 32 mM ammonium formate/acid formic buffer at pH 3.1, with a working voltage of 25 kV. The MS detection of the five pesticides was performed in negative ionization mode. Full-scan spectra with base peaks corresponding to [M-H]- were obtained except for acifluorfen, which gives [M-H-CO2]- as most abundant ion. Compared with the conventional EC-UV, the limits of detection were lower for acifluorfen, haloxyfop, ioxynil, and picloram, by a factor of 20, 20, 50, and 2, respectively. Extraction involved fruit sample homogenization with an acetone-water solution (5:1), filtration, and acetone evaporation prior to fiber extraction. SPME conditions such as time, pH, ion strength, stationary phase of the fiber, sample matrix, and desorption solvents were examined. The recovery of the analytes ranged from 7 to 94%, and the relative standard deviation was between 3 and, 13%. The method was found to be linear between 0.02 and 500 mg kg(-1) with correlation coefficients ranging from 0.992 to 0.997. The limits of quantification were from 0.02 to 5 mg kg(-1). The optimized method was successfully applied to the analysis of acid pesticides in fruit samples. PMID- 12585471 TI - High mass measurement accuracy determination for proteomics using multivariate regression fitting: application to electrospray ionization time-of-flight mass spectrometry. AB - Important factors that limit the mass measurement accuracy from a mass spectrometer are related to (1) the type of mass analyzer used and (2) the data processing/calibration methods used to obtain mass values from the raw data. Here, two data processing methods are presented that correct for systematic deviations when the mass of ions is measured using a time-of-flight (TOF) mass spectrometer. The first fitting method is one where m/z values are obtained from fitting peak distributions using double Gaussian functions. A second calibration method takes into account the slight nonlinear response of the TOF analyzer in addition to the drift in the calibration over time. Using multivariate regression, both of these two effects can be corrected for using a single calibration formula. Achievable performance was evaluated with a trypsin digestion of serum albumin and proteins from the organism D. radiodurans that was analyzed using gradient reversed-phase liquid chromatography combined with an electrospray ionization orthogonal TOF mass spectrometer. The root-mean-square deviation between the theoretical and experimental m/z values for serum albumin tryptic peptides was found to be 8 ppm using the double Gaussian-multivariate method compared to 29 ppm determined using linear calibration and normal peak centroiding. An advantage of the methods presented here is that no calibrant compounds need to be added to the mobile phase, thereby avoiding interference effects and signal suppression of analytes. PMID- 12585472 TI - Study of the fragmentation mechanism of protonated 6-hydroxychlorzoxazone: application in simultaneous analysis of CYP2E1 activity with major human cytochrome P450s. AB - The application of liquid chromatography tandem mass spectrometry for simultaneous analysis of major human cytochrome P450 activities via a single atmospheric pressure ionization (API) LC/MS/MS method has been hampered by the preferred detection of 6-hydroxychlorzoxazone (HCZ), the metabolite of the CYP2E1 probe, chlorzoxazone, under negative API. An initial simulation of the dissociation constants suggested the potential ionization of the enol form of HCZ at low pH, and the accurate mass measurements confirmed the presence of the protonated HCZ signal under (+) ESI at pH 3. However, the CID spectrum of the protonated HCZ resulted in a few intense, but uncommon, fragment ions that could be utilized for specific selected reaction monitoring (SRM) transitions. The deduced elemental compositions of these fragment ions indicated possible aromatic ring opening for the first two intense product ions at m/z 130 and 115, as well as chlorine radical loss for the third ion at m/z 151. Further precursor and product ion scan studies, along with the deuterium ion exchange in solution, revealed the involvement of three distinct pathways of fragmentation. The m/z 186 ->130 transition, which was shown to be specific in human plasma and rat hepatic microsomes, was further combined with the SRM transition of reserpine (internal standard) and eight probe substrates for human cytochrome P450 isoforms. This led to the development of a full LC/MS/MS method capable of analyzing a total of nine human P450 activities within 3 min, including CYP2E1, using a single assay in the (+) ESI mode. The HCZ assay showed excellent linearity with a coefficient of determination (R2) greater than 0.98 at dynamic range of 0.05 (LOQ) to 40 microM. Preliminary data from the three-day validation of the HCZ assay indicated that the accuracy and precision for quality control samples was within +/- 15% of the spiked concentration at all levels. PMID- 12585473 TI - Probing the vapor-liquid phase behaviors of near-critical and supercritical fluids using a shear mode piezoelectric sensor. AB - With the rapidly expanding industrial and research applications of near-critical and supercritical technology there is a pressing need for a simple and inexpensive sensor that may be used to determine the phase coexistence regions of fluid mixtures and to establish whether a fluid system is below, at, or above, a critical point. Mechanically vibrating AT-cut quartz plates may be used to determine the product of the fluid density and viscosity of a fluid in which it is immersed, through measurement of the impedance minimum of the electrical equivalent circuit or of the corresponding frequency. The density-viscosity product changes abruptly between fluid phases and rapidly along the isotherm corresponding to the critical temperature, enabling such a plate to act as a sensor of these fluid features. We consider the limitations and linearity of such a sensor and its behavior when a liquid-gas meniscus crosses its surface. We demonstrate for the first time the effective use of an AT-cut quartz sensor in mapping the phase behavior of fluids, using measurements made on carbon dioxide and ethane for calibration and then investigating an ethane-carbon dioxide mixture. The advantages of this experimental approach are that (i) piezoelectric sensors are available for operation up to 1,000 degrees C and at extremely high pressures and (ii) the measurement of the density-viscosity product of supercritical fluids is inherently simpler than traditional techniques for determining phase behavior. PMID- 12585474 TI - Hydrodynamic study of ion transfer at the liquid/liquid interface: the channel flow cell. AB - A hydrodynamic system based on the channel flow cell for voltammetric detection of ions at the liquid/liquid interface is reported. The current response for tetraethylammonium ion transfer across a membrane-supported liquid/liquid interface is shown to be consistent with existing theory for both the flow rate and voltage scan rate dependence of such processes, with no calibration factors or other adjustable parameters required. The analytical utility of such a device is discussed with specific regard to in situ measurements in flow systems. PMID- 12585475 TI - Nonideal electrochemical behavior of biomimetic iron porphyrins: interfacial potential distribution across multilayer films. AB - The electrochemical behavior of multilayer films formed by iron porphyrins deposited on an edge plane graphite electrode has been examined under anaerobic conditions. In the scan rate interval (1-250 mV/s) where the electrode reaction is reversible, CV diagrams of these films demonstrate substantial deviations from ideality in broadening and separation of the peaks. A model that describes the observed behavior is proposed by taking into account the potential distribution at the electrode/film interface and the concentration dependence of surface activity coefficients. The peak separation is described in terms of the electric double layer that affects the potential difference driving the electrode reaction. The effective potential difference deviates from the applied value due to the potential distribution across the film. The interfacial potential distribution depends on the ionic concentration inside the film. When different ionic concentrations are assumed for oxidation and reduction, different shifts from the applied potential lead to a hysteresis of the peaks. The peak broadening is modeled by using the lattice theory expression for the surface activity coefficients. The model shows that the midpoint potentials of the redox centers depend on the ionic concentration inside the film. At low ionic concentrations, they are remarkably close to the midpoints of the cytochrome c oxidase heme a3/CuB site. PMID- 12585476 TI - A low-flow ce/electrospray ionization MS interface for capillary zone electrophoresis, large-volume sample stacking, and micellar electrokinetic chromatography. AB - A simple and versatile low-flow interface has been developed for interfacing capillary electrophoresis (CE) with electrospray ionization (ESI) mass spectrometry. This low-flow interface showed better sensitivity than a conventional sheath liquid interface, primarily attributed to a low dilution factor and a reduction in the sprayer orifice size. The interface was also found to be more tolerant to the presence of nonvolatile salts. Because of tolerance to the surfactant SDS, this interface can be used to couple micellar electrokinetic chromatography (MEKC) with ESI-MS. The performance of the interface in an MEKC-MS application, as demonstrated in the analysis of triazines, was significantly better than that obtained with a conventional sheath liquid interface. Moreover, this interface can be easily used for large-volume sample-stacking (LVSS) applications. Using a series of phenols as a test case, an approximate 500-fold enrichment was achieved by LVSS in conjunction with the low-flow CE/MS interface described. PMID- 12585477 TI - Analysis of NAD(P)+/NAD(P)H cofactors by imprinted polymer membranes associated with ion-sensitive field-effect transistor devices and Au-quartz crystals. AB - Specific recognition sites for the NAD(P)+ and NAD(P)H cofactors are imprinted in a cross-linked acrylamide-acrylamidophenylboronic acid copolymer membrane. The imprinted membranes, associated with pH-sensitive field-effect transistors (ISFETs) or Au-quartz piezoelectric crystals, enable the potentiometric or microgravimetric analysis of the oxidized NAD(P)+ cofactors and the reduced NAD(P)H cofactors, respectively. The NAD+- and NADP+-imprinted membranes associated with the ISFET allow the analysis of NAD+ and NADP+ with sensitivities that correspond to 15.0 and 18.0 mVdecade(-1) and detection limits of 4 x 10(-7) and 2 x 10(-7) M, respectively. The NADH- and NADPH-imprinted membranes associated with the ISFET device enable the analysis of NADH and NADPH with sensitivities that correspond to 24.2 and 21.8 mV x decade(-1) and lower detection limits that are 1 x 10(-7) and 2 x 10(-7) M, respectively. The ISFET devices functionalized with the NADH and NADPH membranes are employed in the analysis of the biocatalyzed oxidation of lactic acid and ethanol in the presence of lactate dehydrogenase and alcohol dehydrogenase, respectively. PMID- 12585478 TI - Amperometric detection of quantal catecholamine secretion from individual cells on micromachined silicon chips. AB - We have fabricated electrochemical electrodes in picolitersized wells for measuring catecholamine release from individual cells with millisecond resolution. Each well-electrode roughly conforms to the shape of the cell in order to capture a large fraction of released catecholamine with high time resolution. Using this device, we can resolve spikes in amperometric current corresponding to quantal catecholamine release via exocytosis. In addition, we have combined amperometric recording on the chip with patch-clamp recordings of membrane capacitance as an assay of exocytosis. A quantitative comparison of the two methods suggests that a large fraction of catecholamine release is oxidized on the surface of the well-electrode. This technology has applications in cell based biosensor development, high-throughput screening of drugs, and basic science investigations. PMID- 12585479 TI - Microfabricated electrophoresis chip for bioassay of renal markers. AB - A novel capillary electrophoresis chip-based detection system for simultaneous measurements of the renal markers creatine, creatinine, p-aminohippuric acid, and uric acid is described. Fluid control is used for mixing the sample with the enzymes creatininase (CA), creatinase (CI), and sarcosine oxidase (SOx) and for separating the neutral hydrogen peroxide end product from the anionic p aminohippuric and urate species. The 'total' (creatinine and creatine) signal was measured with the running buffer containing all three enzymes, while the creatine signal alone was recorded by mixing the sample with the CI-SOx solution. Creatinine concentrations are measured by comparing the response in the presence and absence of CA. The peroxide product and the oxidizable p-aminohippuric and uric acids are detected electrochemically at a downstream gold-coated thick-film amperometric detector. The four renal markers are readily measured within 5 min, while creatinine/creatine within less than 2 min. Factors influencing the performance, including the level of three enzymes, separation voltage, and detection potential, are optimized. Applicability to urine samples is demonstrated. Such a multianalyte microchip detection device would allow renal function testing to be performed more rapidly, easily, and economically in the point-of-care setting. PMID- 12585480 TI - Application of diamond microelectrodes for end-column electrochemical detection in capillary electrophoresis. AB - Highly boron-doped diamond microelectrodes were employed in an end-column electrochemical detector for capillary electrophoresis (CE). The diamond microline electrodes were fabricated from conducting diamond thin films (exposed surface area, 300 x 50 microm), and their analytical performance as CE detectors was evaluated in a laboratory-made CE installation. The CE-ED system exhibited high separation efficiency for the detection of several catecholamines, including dopamine (DA), norepinephrine (NE), and epinephrine (E), with excellent analytical performance, for example, 155,000 theoretical plates for DA. The diamond-based electrochemical detection system also displayed low detection limits (approximately 20 nM for E at S/N = 3) and a highly reproducible current response with 10 repetitive injections of mixed analytes containing DA, NE, and E (each 50 microM), with relative standard deviations (RSD) of approximately 5%. The performance of the diamond detector in CE was also evaluated in the detection of chlorinated phenols (CP). When compared to the carbon fiber microelectrode, the diamond electrode exhibited lower detection limits in an end-column CE detection resulting from very low noise levels and highly reproducible analyses without electrode polishing due to analyte fouling, which makes it possible to perform easier and more stable CE analysis. PMID- 12585481 TI - Enantioselective hydrolysis of some 2-aryloxyalkanoic acid methyl esters and isosteric analogues using a penicillin G acylase-based HPLC monolithic silica column. AB - A technique based on liquid chromatography has been developed to facilitate studies of enantioselectivity in penicillin G acylase (PGA)-catalyzed hydrolysis of some 2-aryloxyalkanoic acid methyl esters and isosteric analogues. PGA was covalently immobilized on an aminopropyl monolithic silica support to create an immobilized HPLC-enzyme reactor. Two sets of experimental data were drawn to calculate the enantioselectivity (E) of the kinetically controlled enantiomer differentiating reaction, the degree of substrate conversion and the enantiomeric excess of the product. The developed enzymatic reactor was coupled through a switching valve to an achiral analytical column for separation and quantitation of the hydrolysis products. The enantiomeric excess was determined off-line on a PGA-chiral stationary phase. In this way, highly precise E values were determined. A computational study related to the hydrolysis of the considered racemic esters was also carried out in order to unambiguously clarify both the substrate specificity and the enantioselectivity displayed by PGA. PMID- 12585482 TI - Directed immobilization of peptide ligands to accessible pore sites by conjugation with a placeholder molecule. AB - When small ligands are immobilized onto a porous chromatography medium, only a limited number of binding sites contributes to the interaction with the target molecule. The main part of the ligand molecules is distributed on sites that are not accessible for the target protein due to steric hindrance. To direct the ligand into a well-accessible position, the ligand was conjugated to a large molecule that acted as a placeholder during the immobilization step. Then the placeholder molecule was cleaved off and washed out. Two linear peptides with affinity for lysozyme and human blood coagulation factor VIII, respectively, were studied as model systems. The protected peptide ligand was covalently linked to a 20-kDa poly(ethylene glycol) molecule containing an acid-labile linker. After selective deprotection of the peptide and purification, immobilization of this conjugate on a preactivated chromatography matrix was performed alternatively through the free N-terminus, the epsilon-amino group of lysine, or the sulfohydryl group of cysteine. After the immobilization reaction, the spacer molecule and remaining protecting groups were cleaved off and the gels were tested by affinity chromatography. This novel immobilization technique substantially increased the binding capacity and the ligand utilization for the target protein, and site-specific immobilization could be demonstrated. PMID- 12585483 TI - A fluorescent chemosensor for sodium based on photoinduced electron transfer. AB - A new optical sensor suitable for practical measurement of sodium in serum and whole blood samples is described. The optical sensor is based on a novel PET (photoinduced electron transfer) fluoroionophore immobilized in a hydrophilic polymer layer. The design concept of the fluoroionophore follows the receptor spacer-fluorophore approach to sensor design using intramolecular PET-based signal transduction. Key to the development of this sensor is the identification of a nitrogen-containing, sodium-binding ionophore, coupled with a fluorophore having the correct spectral and electron-accepting properties. The slope of the sensor is approximately 0.5%/mM in the typical clinically significant range of 120-160 mM. This sensor has been implemented into a disposable cartridge, used for a commercially available critical care analyzer (Roche OPTI CCA) with precision better than +/- 1 mM (1 SD). The sensor displays excellent stability against hydrolysis and oxidation, leading to slope changes <5% after 9 months wet storage at 30 degrees C. On the basis of this design concept, fluoroionophores for other cations such as potassium, calcium and magnesium can be prepared by substitution of the ionophore. PMID- 12585484 TI - Microarrays for the screening of allergen-specific IgE in human serum. AB - The described in vitro test system for allergy diagnosis is based on microscope glass slides activated with (3-glycidyloxypropyl)trimethoxysilane. Allergen solutions are immobilized as small droplets (approximately 10 nL) on the activated glass slides with a piezoelectric arrayer. In contrast to other tests for specific IgE, such as Pharmacia CAP FEIA, AlaSTAT, or FAST, only a 25-microL serum sample is needed for the screening of allergen-specific IgE against a multitude of allergens and the test can be performed in less than 1 h. Compared with multiallergen dipstick screening tests (e.g., IgEquick, CMG Immunodot) based on multiallergen-coated nitrocellulose strips, the measurement of the microarray based system can be performed automatically. The chemiluminescence intensities are detected with a sensitive CCD camera. Allergen extracts and recombinant/purified allergens (24 preparations) have been used on the same modified surface for the screening of allergen-specific IgE. With these disposable microarray slides, it is possible to distinguish between patients with and without elevated levels of allergen-specific IgE. Repeated measurements of serum samples demonstrated a sufficient reproducibility. Detection limits (microg/L) of 0.35 (r Betvl), 0.16 (PLA2), and 1.9 (Der p1) were achieved. PMID- 12585485 TI - Scanning electrochemical microscopy of model neurons: imaging and real-time detection of morphological changes. AB - Living PC12 cells, a model cell type for studying neuronal function, were imaged using the negative feedback mode of a scanning electrochemical microscope (SECM). Six biocompatible redox mediators were successfully identified from a large pool of candidates and were then used for imaging PC12 cells before and after exposure to nerve growth factor (NGF). When exposed to NGF, cells differentiate into a neuron phenotype by growing narrow neurites (1-2 microm wide) that can extend > 100 microm from the cell proper. We demonstrate that carbon fiber electrodes with reduced tip diameters can be used for imaging both the cell proper and these neurites. Regions of decreased current, possibly resulting from raised features not identifiable by light microscopy, are clearly evident in the SECM images. Changes in the morphology of undifferentiated PC12 cells could be detected in real time with the SECM. After exposure to hypotonic and hypertonic solutions, reversible changes in cell height of <2 microm were measured. PMID- 12585486 TI - Development and characterization of microfabricated disposable gold working electrodes for high-performance ion chromatography and integrated pulsed amperometric detection. AB - We have developed a new type of microfabricated thin-film electrode on polymeric substrates. The microfabrication process allows for inexpensive and reproducible mass production of disposable working electrodes for high-performance ion chromatography and integrated pulsed amperometric detection (IPAD). These microfabricated electrodes are disposable and have been optimized for use in flow through low-dead-volume electrochemical cells. The analytical performance of microfabricated gold electrodes was characterized with the help of the IPAD method for amino acid detection under alkaline conditions required for anion exchange separations. When used with a new optimized six-potential IPAD waveform, the electrodes functioned properly for weeks. Compared to nondisposable working electrodes, the disposable working electrodes generated equal or better results in the limit of detection, linearity of calibration, and reproducibility. Disposable electrodes make it possible to avoid polishing and reconditioning, which are required with nondisposable electrodes. PMID- 12585487 TI - Combined phage typing and amperometric detection of released enzymatic activity for the specific identification and quantification of bacteria. AB - Here, we describe a novel electrochemical method for the rapid identification and quantification of pathogenic and polluting bacteria. The design incorporates a bacteriophage, a virus that recognizes, infects, and lyses only one bacterial species among mixed populations, thereby releasing intracellular enzymes that can be monitored by the amperometic measurement of enzymatic activity. As a model system, we used virulent phage typing and cell-marker enzyme activity (beta-D galactosidase), a combination that is specific for the bacterial strain Escherichia coli (K-12, MG1655). Filtration and preincubation before infecting the bacteria with the phage enabled amperometric detection at a wide range of concentrations, reaching as low as 1 colony-forming unit/100 mL within 6-8 h. In principle, this electrochemical method can be applied to any type of bacterium phage combination by measuring the enzymatic marker released by the lytic cycle of a specific phage. PMID- 12585488 TI - An analogy of an ion-selective electrode sensor based on the voltammetry of microcrystals of tetracyanoquinodimethane or tetrathiafulvalene adhered to an electrode surface. AB - The voltammetry of solid 7,7,8,8-tetracyanoquinodimethane (TCNQ) and tetrathiafulvalene (ITF) at an electrode-microparticle-aqueous (electrolyte) interface generates characteristic current-potential profiles associated with solid-solid-phase transformations. During the reactions, electrolyte ions are included into the TCNQ (cations) and TTF (anions) lattice sites as part of the charge neutralization process. Consequently, electrolyte ion concentration is associated with the reversible potential of the TCNQ0/- and TTF0/+ reactions, making these processes candidates for the development of novel voltammetric cation and anion sensors, respectively. Electrode potential-analyte ion concentration dependence studies exhibited highly reproducible potential shifts of 45 (+/- 1) mV/decade change in ion analyte concentration for both the TCNQ cation sensor and the TTF anion sensor. When presented with mixed-analyte solutions, both ion-sensing systems exhibited a degree of ion selectivity. Ion selectivity trends may be modeled using equations based on a Nicolsky-type selectivity relationship, in accordance with the concept that these are the voltammetric analogies of potentiometric ion-selective membrane electrodes. PMID- 12585489 TI - Solution to the problem of interferences in electrochemical sensors using the fill-and-flow channel biosensor. AB - A generic fill-and-flow channel biosensor with upstream electrodes to determine the extent of interferences in the sample is described. A pair of upstream electrodes poised at a suitable potential allows both the calculation of the extent of removal of interfering agents and the effect of interfering agents at the detector electrode. A model was developed and tested that predicts the concentrations of all species throughout the channel and, hence, the current at each electrode due to each species. This enables correction of the detector electrode current and a more accurate determination of the analyte concentration. The concept was applied to a biosensor for the determination of glucose in the presence of ascorbic acid, acetamidophenol, and uric acid, as well as glucose in wine samples containing polyphenolic interfering agents. PMID- 12585490 TI - Effects of poly(ethylene glycol) tert-octylphenyl ether on tris(2 phenylpyridine)iridium(III)-tripropylamine electrochemiluminescence. AB - The effects of the nonionic surfactant Triton X-100 (poly(ethylene glycol) tert octylphenyl ether) on the properties of tris(2-phenylpyridine)iridium(III) (Ir(ppy)3, where ppy = 2-phenylpyridine, electrochemiluminescence (ECL) have been investigated. Anodic oxidation of Ir(ppy)3 produces ECL in the presence of tri-n propylamine (TPrA) in aqueous surfactant solution. Increases in ECL efficiency (> or = 10-fold) and TPrA oxidation current (> or = 2.0-fold) have been observed in surfactant media. The data support adsorption of surfactant on the electrode surface, thus facilitating TPrA and Ir(ppy)3 oxidation and leading to higher ECL efficiencies. PMID- 12585491 TI - Characterization and quantitation of aprepitant drug substance polymorphs by attenuated total reflectance fourier transform infrared spectroscopy. AB - In this study, we report the use of attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FT-IR) for the identification and quantitation of two polymorphs of Aprepitant, a substance P antagonist for chemotherapy-induced emesis. Mixtures of the polymorph pair were prepared by weight and ATR-FT-IR spectra of the powdered samples were obtained over the wavelength range of 700-1500 cm(-1). Significant spectral differences between the two polymorphs at 1140 cm(-1) show that ATR-FT-IR can provide definitive identification of the polymorphs. To investigate the feasibility of ATR-FT-IR for quantitation of polymorphic forms of Aprepitant, a calibration plot was constructed with known mixtures of the two polymorphs by plotting the peak ratio of the second derivative of absorbance spectra against the weight percent of form II in the polymorphic mixture. Using this novel approach, 3 wt % of one crystal form could be detected in mixtures of the two polymorphs. The accuracy of ATR-FT IR in determining polymorph purity of the drug substance was tested by comparing the results with those obtained by X-ray powder diffractometry (XRPD). Indeed, polymorphic purity results obtained by ATR-FT-IR were found to be in good agreement with the predictions made by XRPD and compared favorably with actual values in the known mixtures. The present study clearly demonstrates the potential of ATR-FT-IR as a quick, easy, and inexpensive alternative to XRPD for the determination of polymorphic identity and purity of solid drug substances. The technique is ideally suited for polymorph analysis, because it is precise, accurate, and requires minimal sample preparation. PMID- 12585492 TI - Porphyrin assembly on beta-cyclodextrin for selective sensing and detection of a zinc ion based on the dual emission fluorescence ratio. AB - In the present paper, a new cyclodextrin/porphyrin supramolecular sensitizer for zinc ion has been proposed based on the porphyrin dual fluorescence emission ratio. In aqueous solution, meso-tetraphenylporphyrin shows weak fluorescence, while in the presence of alkylated beta-cyclodextrin, it exhibits significant fluorescence enhancement by forming a cyclodextrin/porphyrin inclusion complex. Furthermore, the formation of a supramolecular complex causes a remarkable increase of the porphyrin metalation rate following the porphyrin fluorescence emission changes at two different emission wavelengths. The fluorescence emission of tetraphenylporphyrin at 656-nm bands decreases while that at 606 nm increases upon zinc ion interaction. Thus, the inclusion complex can behave as a ratiometric fluorescent sensor. Theoretically derivative equations for fluorescent ratiometry have been proposed for the first time. The feasibility of the proposed method is demonstrated by the performance of fluorometric detection of zinc ion. With the optimum conditions described, zinc ion in aqueous solution can be determined from 5.0 x 10(-7) to 2.5 x 10(-4) M. As the porphyrin electronic absorption and fluorescence emission are located in the visible range, and the fluorescence changes upon zinc ion interaction show high selectivity over biologically relevant cations, the inclusion complex could be used for biomedical application. PMID- 12585493 TI - Reaction monitoring of aliphatic amines in supercritical carbon dioxide by proton nuclear magnetic resonance spectroscopy and implications for supercritical fluid chromatography. AB - In the recent years, it has repeatedly been stated that amines react with CO2 and can therefore not be chromatographed under supercritical conditions with CO2. The aim of the present work is to elucidate the structural requirements and conditions that can lead to the reaction of an amine analyte with CO2 and, if this occurs, the structure of the formed product. The use of on-line nuclear magnetic resonance (NMR) spectroscopy with a flow probe for supercritical fluid chromatography (SFC) enables the investigation of these unstable analytes in supercritical mediums. Several alkyl-substituted secondary benzylamines and some primary aromatic amines were dissolved in supercritical CO2 and investigated by employing on-line SFC-1H NMR spectroscopy. It was found that the condition of carbamic acid formation depends on the steric properties of the substituents of the amine. A 2-isopropylamino alcohol compound, metoprolol, was also investigated with the setup. No carbamic acid could be detected with the present conditions. PMID- 12585494 TI - Transferred nuclear overhauser effect in nuclear magnetic resonance diffusion measurements of ligand-protein binding. AB - The drug discovery process relies on characterizing structure-activity relationships, since specific ligand-target interactions often result in important biological functions. Measuring diffusion coefficients by nuclear magnetic resonance spectroscopy is a useful way to study binding, because changes can be detected when a small ligand interacts with a macromolecular target. Diffusion coefficients can be miscalculated, however, due to magnetization transfer between the receptor and ligand. This transferred nuclear Overhauser effect (trNOE) disrupts the observed signal decay due to diffusion as a function of the experimental diffusion time. Since longer diffusion times also selectively edit free ligand signal, the measured diffusion coefficients become biased toward the fraction of bound ligand. Despite this discrepancy, under these experimental conditions, the trNOE selectively influences the measured signals of binding ligands and can be used to gain insight into ligand-protein interactions. These phenomena have been studied for caffeine and L-tryptophan, which bind to human serum albumin, and the antimalarial agent trimethoprim, which interacts with dihydrofolate reductase. The results provide insight into the nature of ligand protein binding and are thus useful for elucidating the molecular features of the ligand that interact with the protein. PMID- 12585495 TI - Generation of atomic and molecular cadmium species from aqueous media. AB - Generation of atomic and molecular species of cadmium following reaction of acidified sample with sodium tetrahydroborate reductant using a modified parallel path Burgener nebulizer has been accomplished. Once generated, atomic species are stable in water, having a half-life of 2.2 min, and can be preconcentrated for subsequent quantitation. Although the initial products of the reduction reaction appear to be free atoms, subsequent interaction with nascent hydrogen gives rise to the molecular hydride in a transformation moderated by the chemistry of surface groups exposed to the reaction products. Use of a glass spray chamber results in a predominance of atomic vapor whereas a polymeric spray chamber releases the volatile molecular species. PMID- 12585496 TI - Application of a combination of hard and soft modeling for equilibrium systems to the quantitative analysis of pH-modulated mixture samples. AB - pH modulation of aqueous mixture samples combined with FT-IR detection and a powerful second-order resolution method is proposed for both resolution and quantitation of acid analytes in the presence of similarly behaving interferences. The proposed method allows for the analyte determination in mixtures using a single standard sample per analyte. Due to the very similar pKa values of the investigated analytes and interferences, the highly correlated concentration profiles of these compounds cannot be successfully resolved with pure soft-modeling second-order approaches. The inclusion of a hard-modeling constraint based on the acid-base equilibrium model in the soft-modeling curve resolution method has allowed for the unambiguous resolution of the analyte profiles and, as a consequence, for the correct quantitation of this compound in the mixture sample. A detailed discussion of the combined hard-soft-modeling approach as well as the analytical problem and the quantitation results is given. Also, strategies to overcome problems associated with variation in pKa values between different samples are addressed. Due to the flexible implementation of the hard-model equilibrium constraint in the multivariate curve resolution alternating least squares method, this approach is expected to be useful also for analysis of other complex mixed equilibrium-based chemical systems. PMID- 12585497 TI - A high repetition rate (1 kHz) microcrystal laser for high throughput atmospheric pressure MALDI-quadrupole-time-of-flight mass spectrometry. AB - Sample throughput has been increased in many areas of proteomics, but the last significant advance in lasers used for matrix-assisted laser desorption/ionization (MALDI) was the introduction of cartridge-type N2 lasers (337 nm, 4-ns pulse widths, 1-30-Hz repetition rates) more than a decade ago. This report describes the application of a 1-kHz repetition rate Nd:YAG laser (355 nm, <500-ps pulse widths) for atmospheric pressure MALDI-QqTOFMS, and data obtained are compared to a conventional nitrogen laser. For example, the signal intensity for angiotensin II using the 1-kHz laser was in some cases enhanced by a factor of 80 and high-quality data could be obtained in as little as 1 s. PMID- 12585498 TI - Flavor release measurement by atmospheric pressure chemical ionization ion trap mass spectrometry, construction of interface and mathematical modeling of release profiles. AB - An instrumental on-line retronasal flavor analysis was developed to obtain information about the release of flavor compounds in expired air from humans during eating. The volatile flavor compounds were measured by ion trap mass spectrometry with an atmospheric pressure chemical ionization source (APCI). An interface was designed to sample the breath directly from the nose. The repeatability in vitro for seven different flavor compounds came out with relative standard derivation less than 10% in most cases, which is acceptable. In vitro quantification was carried out by a determination of the concentration in the gas phase over a flavor solution by GC/MS, followed by measurements of intensities by the APCI ion trap. Ion suppression by acetone in the breath was negligible at concentration levels relevant in these experiments. The instrumental limits of detection for menthone and menthol coincide with that of the flavor detection threshold. An application study on the release of menthone and menthol from chewing gum by a group of six test persons was performed. Flavored chewing gum was used as a model matrix because of the long chewing periods and the simplicity of the system. It is concluded that the interface and the method can be used to measure breath from the nose. A mathematical model of the data was developed to give a quantitative method for description and characterization of the release of flavor compounds. The release profiles consisted of two sequences, one for a chewing period, and one for a phasing out process. The proposed method for modeling provided a reasonable description of the release process. In addition to flavor compounds, this new interface and mathematical application could provide information on chemicals in the human breath, which could be interesting, for example, within medical diagnosis. PMID- 12585499 TI - Stop and go extraction tips for matrix-assisted laser desorption/ionization, nanoelectrospray, and LC/MS sample pretreatment in proteomics. AB - Proteomics is critically dependent on optimal sample preparation. Particularly, the interface between protein digestion and mass spectrometric analysis has a large influence on the overall quality and sensitivity of the analysis. We here describe a novel procedure in which a very small disk of beads embedded in a Teflon meshwork is placed as a microcolumn into pipet tips. Termed Stage, for STop And Go Extraction, the procedure has been implemented with commercially available material (C18 Empore Disks (3M, Minneapolis, MN)) as frit and separation material. The disk is introduced in a simple and fast process yielding a convenient and completely reliable procedure for the production of self-packed microcolumns in pipet tips. It is held in place free of obstacles solely by the narrowing tip, ensuring optimized loading and elution of analytes. Five disks are conveniently placed in 1 min, adding <0.1 cent in material costs to the price of each tip. The system allows fast loading with low backpressure (>300 micro/min for the packed column using manual force) while eliminating the possibility of blocking. The loading capacity of C18-StageTips (column bed: 0.4 mm diameter, 0.5 mm length) is 2-4 microg of protein digest, which can be increased by using larger diameter or stacked disks. Five femtomole of tryptic BSA digest could be recovered quantitatively. We have found that the Stage system is well-suited as a universal sample preparation system for proteomics. PMID- 12585500 TI - Complexation of copper by zwitterionic aminosulfonic (good) buffers. AB - Copper binding properties were investigated for several popular zwitterionic buffers. The two buffers 4-morpholinoethanesulfonic acid (MES) and 3-N morpholinopropanesulfonic acid (MOPS) did not bind copper and would be good choices for metal speciation studies within their operational pH range. Conversely, 3-(N-morpholino)-2-hydroxypropanesulfonic acid (MOPSO) was observed to weakly bind copper directly (log Kc 2.02). Moreover, strong copper binding was observed for 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 4-(2 hydroxyethyl)piperazine-1-propanesulfonic acid (HEPPS), and N-(2 hydroxyethyl)piperazine-N'-(2-hydroxypropanesulfonic acid) (HEPPSO). Log Kc values range from 7.04 to 7.68 and are indicative of strong copper binding ligands. The latter buffer also exhibited weak binding characteristics with a log Kc of 2.05. The strong Cu binding ligands were present in HEPES, HEPPS, and HEPPSO at much lower concentrations than the total buffer concentration. MES, HEPES, MOPSO, and HEPPSO were analyzed by electrospray-ionization quadrapole time of-flight mass spectroscopy. The most prominent feature of the spectra for each buffer analyzed was the presence of multiple oligomers, indicating a propensity of interaction between buffer molecules. In addition, the presence of several contaminants was identified in the mass spectrum of the HEPES matrix, including a prominent contaminant (at m/z 131) present in levels similar to those obtained from the modeling of the copper titration data. Other contaminants were found in the other matrixes but were not identified as possible copper binding agents. PMID- 12585501 TI - Chiral analysis using the kinetic method with optimized fixed ligands: applications to some antibiotics. AB - A new version of the kinetic method for chiral analysis, which employs a fixed (nondissociating) ligand as well as the usual analyte and chiral reference ligands, is introduced to simplify the kinetics of this experiment. Singly charged clusters containing the divalent transition metal ion MnII, a peptide which serves as a fixed ligand, an amino acid chiral reference, and the analyte 4 benzyl-2-oxazolidinone were generated by electrospray ionization (ESI). The cluster ion of interest was mass-selected, and the kinetics of its competitive unimolecular dissociations was investigated in an ion trap mass spectrometer. The chiral selectivity (R(fixed)chiral), the ratio of the two fragment ion abundances when the cluster contains one pure enantiomer of the analyte expressed relative to that for the other enantiomer, varies with increasing size of the fixed peptide ligands. The metal-ligand and the ligand-ligand interactions that produce chiral discrimination are optimized in the tetrapeptide fixed ligand Gly-Gly-Ala Gly, as shown by data for 15 fixed ligands. The difference in the free energies of activation for the two competitive reactions is estimated to be approximately 7 kJ/mol for this particular fixed ligand. The sensitive nature of the methodology and the linear relationship between the logarithm of the fragment ion abundance ratio and the optical purity (intrinsic to the kinetic method) allows mixtures to be analyzed for as little as 1% enantiomeric excess (ee), by simply recording the ratios of fragment ion abundances in a tandem mass spectrum. These features are demonstrated in the case of the pharmacologically important 4-benzyl 2-oxazolidinones and in the case of penicillamine. PMID- 12585502 TI - Determination of natural and synthetic colorants in prescreened dairy samples using liquid chromatography-diode array detection. AB - A simple and novel screening method for food colorants was proposed. Synthetic or natural colorants were discriminated as they were selectively adsorbed on cotton or RP-C18 sorbent columns, respectively. After elution, each fraction was monitored at 400, 530, and 610 nm for yellow, red, and green-blue-brown additives, respectively, with a DAD spectrophotometer. The screening method serves as a filter to indicate whether the target colorants are present above or below the detection limit of the method (6-15 or 25-10,000 n/mL for synthetic or natural colorants, respectively). Positive samples were discriminated by LC-DAD, using a flow system similar to that of the screening method. The LC-DAD discrimination/confirmation method is very sensitive; it exhibits a linear range of 0.01-50 microg/mL (excluding curcumin and caramel, which are linear up to 200 and 1,500 microg/mL, respectively). The method was applied to the determination of natural and synthetic colorants in dairy samples with good precision. PMID- 12585503 TI - Use of bioactive glass slides for matrix-assisted laser desorption/ionization analysis: application to microorganisms. AB - Glass slides are widely used in high-throughput analysis and are available commercially with surfaces activated, etched, and channeled. Thin glass microscope slides are shown here to be suitable sample supports for matrix assisted laser desorption/ionization (MALDI) mass spectrometry. As a demonstration, lectins immobilized on glass slides with activated surfaces are used to concentrate and purify agglutinated Bacillis spores. It is expected that such slides will provide a rapid, inexpensive way to evaluate and implement new strategies involving MALDI MS readout. PMID- 12585504 TI - Improvement in analysis of O2 in N2 by using Ar drift gas in an ion mobility spectrometer. AB - Ion mobility spectrometry (IMS) is capable of providing real-time analysis of several impurities in inert gases. Because of its poor peak resolution, the IMS is susceptible to peak interferences. Previous study has shown that the detection limit and accuracy of O2 measurement in N2 suffers from peak interference. In many applications, the interference can be avoided by selecting a proper drift gas. For N2 analysis, we have demonstrated that the peak interference of O2 can be eliminated by using Ar as a drift gas. The use of Ar changes the chemical reactions that occur inside the drift region and results in different product ions. The peaks of new product ions are well-separated and do not coincide with the peak representing O2. PMID- 12585505 TI - Determination of trace perchlorate in high-salinity water samples by ion chromatography with on-line preconcentration and preelution. AB - A simple, automated system for the determination of trace perchlorate by ion chromatography (IC) with an online preconcentration technique is reported. The sample is preconcentrated, and less strongly held ions preeluted before the analyte is transferred to the principal separation system. This approach provides low limits of detection (LOD) and is particularly robust toward the effect of high concentrations of common anions, such as those present in groundwater samples. It compares favorably with currently promulgated EPA method 314.0. The LOD (S/N = 3) is 0.77 microg/L for a 2-mL reagent water sample and decreases more or-less proportionately with increasing sample volume, at least up to 20 mL. Even with a sample of conductivity 14.7 mS/cm (approximately that of 0.1 M Na2SO4), the recovery of added perchlorate at the 25.0 microg/L level was still 92%. The concentration of added perchlorate in the range of 1-400 microg/L was linearly correlated to the peak area, with an r2 value of 0.9997. The recovery of perchlorate from artificial samples with different conductivity by the present method compares favorably with those from the currently recommended EPA Method. The ability of this approach to remove matrix interferences suggests that it would be also promising for perchlorate analysis in other challenging samples. PMID- 12585506 TI - Respiratory effects of toluene diisocyanate in the workplace: a discussion of exposure-response relationships. AB - Toluene diisocyanate (TDI) is an important industrial intermediate used in manufacturing flexible polyurethane (PUR) foams, surface coatings, cast elastomers, sealants, and adhesives. In this review long-term trends in workplace exposures to TDI are assessed in both the producing and using industries, and respiratory health effects of TDI are evaluated in relation to workplace TDI concentrations. The key respiratory health effects associated with repeated or long-term TDI exposure are bronchial asthma and an accelerated rate of decline in lung function. In the early years of the industry, annual incidence rates of occupational asthma (OA) due to TDI ranged from 1% to as high as 5 to 6%, depending on the extent of engineering and work practice controls in the various workplaces. Since the mid-1970s, annual OA incidence rates have been <1%, where 8 h TDI concentrations have been maintained below 5 ppb as determined by personal monitoring, even where short-termTDI concentrations above 20 ppb and less frequently above 40 ppb were routinely detected. In these latter settings, there is evidence that the majority of OA cases may be attributable to TDI concentrations well above 20 ppb associated with overexposure incidents. Further study is needed regarding the role of such incidents in inducing respiratory sensitization. Cross-sectional and longitudinal studies of lung function have indicated that continued exposure after development of work-related respiratory symptoms can lead to transient or accelerated fixed declines in forced expiratory volume in 1 sec (FEV1). These findings are congruent with the FEV1 declines demonstrated in general population studies of persons with persistent bronchial hyperresponsiveness or nonoccupational asthma. More recent longitudinal studies in settings with ongoing medical surveillance have provided no consistent evidence of accelerated FEV1 loss among employees exposed up to 5 ppb TDI on an 8 h time-weighted average basis. PMID- 12585507 TI - Health effects of welding. AB - Many of the epidemiology studies performed are difficult to compare because of differences in worker populations, industrial settings, welding techniques, duration of exposure, and other occupational exposures besides welding fumes. Some studies were conducted in carefully controlled work environments, others during actual workplace conditions, and some in laboratories. Epidemiology studies have shown that a large number of welders experience some type of respiratory illness. Respiratory effects seen in full-time welders have included bronchitis, airway irritation, lung function changes, and a possible increase in the incidence of lung cancer. Pulmonary infections are increased in terms of severity, duration, and frequency among welders. Although epidemiological studies have demonstrated an increase in pulmonary illness after exposure to welding fumes, little information of the causality, dose-response, and possible underlying mechanisms regarding the inhalation of welding fumes exists. Even less information is available about the neurological, reproductive, and dermal effects after welding fume exposure. Moreover, carcinogenicity and short-term and long term toxicology studies of welding fumes in animals are lacing or incomplete. Therefore, an understanding of possible adverse health effects of exposure to welding fumes is essential to risk assessment and the development of prevention strategies and will impact a large population of workers. PMID- 12585508 TI - Using the theory of reasoned action to predict organizational misbehavior. AB - A review of literature on organizational behavior and management on predicting work behavior indicated that most reported studies emphasize positive work outcomes, e.g., attachment, performance, and satisfaction, while job related misbehaviors have received relatively less systematic research attention. Yet, forms of employee misconduct in organizations are pervasive and quite costly for both individuals and organizations. We selected two conceptual frameworks for the present investigation: Vardi and Wiener's model of organizational misbehavior and Fishbein and Ajzen's Theory of Reasoned Action. The latter views individual behavior as intentional, a function of rationally based attitudes toward the behavior, and internalized normative pressures concerning such behavior. The former model posits that different (normative and instrumental) internal forces lead to the intention to engage in job-related misbehavior. In this paper we report a scenario based quasi-experimental study especially designed to test the utility of the Theory of Reasoned Action in predicting employee intentions to engage in self-benefitting (Type S), organization-benefitting (Type O, or damaging (Type D) organizational misbehavior. Results support the Theory of Reasoned Action in predicting negative workplace behaviors. Both attitude and subjective norm are useful in explaining organizational misbehavior. We discuss some theoretical and methodological implications for the study of misbehavior intentions in organizations. PMID- 12585509 TI - Intrinsic religiosity and aggression in a sample of intercollegiate athletes. AB - Using self-report questionnaires, ratings of intrinsic religiosity were negatively correlated with those on aggressive attitudes and verbal aggression of 105 intercollegiate athletes. PMID- 12585510 TI - Relations of the Arabic Type A behavior scale with measures of optimism and pessimism. AB - This study investigated the relation of Type A behavior dimensions and measures of optimism and pessimism. The sample of 322 undergraduate students were 80 men and 242 women at Kuwait University. Analysis showed that five dimensions of Type A behavior, accuracy and persistence, speed and time pressure, desire for promotion, seriousness and organization, and workaholism were positively correlated with optimism, while aggression was negatively correlated with optimism and positively correlated with pessimism. The multiple regression analysis indicated that speed and time pressure and desire for promotion contributed significantly to the prediction of optimism, while aggression predicted pessimism more than any other dimension of Type A behavior. PMID- 12585511 TI - Discriminant validity of the Eating Attitudes Test according to American Psychiatric Association and World Health Organization criteria of eating disorders. AB - The purpose of this study was to assess the sensitivity, the specificity, and the positive predictive value of the Eating Attitudes Test in a sample of Spanish nonclinical 18-yr.-olds. 304 subjects answered the Eating Attitudes Test-40, 290 of whom were interviewed individually with the Spanish version of the Schedules for Clinical Assessment in Neuropsychiatry. Eating disorders were diagnosed using ICD-10 and DSM-III-R criteria. The prevalence of eating disorders was higher for ICD-10 (5.2%) than for DSM-III-R (2.6%) and only affected the rate of diagnosis in women. According to ICD-10 criteria, the cut-off of 25 was more sensitive (87.5%) than the cut-off of 30 (75%) and varied little in specificity (93.9% vs 97.1%). The positive predictive value of the Eating Attitudes Test cut-off of 30 for eating disorders (ICD-10) was 36%. Our results support the test as useful for identifying eating disturbances in 18-yr.-olds and suggest assessment of a cut off lower than 30 may be appropriate in the general population if confirmed in further research with a representative sample of adults. PMID- 12585512 TI - Compliance with the item limit of the food supermarket express checkout lane: another look. AB - A total of 68 15-min. observations of customers' behavior at a food supermarket suggests that only about 7% of shoppers observe the item limit of the express lane. The averages tended to be about four pieces. PMID- 12585513 TI - Work experiences and psychological well-being of former hospital-based nurses now employed elsewhere. AB - Hospitals in Ontario have undergone significant restructuring and downsizing during the past few years. Such changes have created negative reactions among both those who survive the downsizing and those who lose their jobs involuntarily. This study compared work satisfactions and indicators of psychological well-being of 36 nurses during these changes and three years later when they were employed elsewhere. Job satisfaction and psychological well-being increased while psychological burnout decreased in this sample. PMID- 12585514 TI - Rasch scaling paranormal belief and experience: structure and semantics of Thalbourne's Australian Sheep-Goat Scale. AB - Research on the relation between demographic variables and paranormal belief remains controversial given the possible semantic distortions introduced by item and test level biases. We illustrate how Rasch scaling can be used to detect such biases and to quantify their effects, using the Australian Sheep-Goal Scale as a substantive example. Based on data from 1.822 respondents, this test was Rasch scalable, reliable, and unbiased at the test level. Consistent with other research in which unbiased measures of paranormal belief were used, extremely weak age and sex effects were found (partial eta2 = .005 and .012, respectively). PMID- 12585515 TI - Changes in the temporal organization of autobiographical memories. AB - The temporal organization of autobiographical memories was examined through the event-cueing technique. Participants were presented a word indicating a life-time period and asked to recall autobiographical events sequentially. Temporal relations between recalled events were analyzed. Analysis suggested that recent autobiographical memories are organized temporally. As time passes, however, organization becomes ambiguous, and old autobiographical memories can evoke temporally distant memories sharing the person, life theme, or other distinctive features. PMID- 12585516 TI - Effects of laughing, smiling, and howling on mood. AB - This study examined the effects of forced laughter on mood and compared laughter with two other possible mood-improving activities, smiling and howling. While howling did not substantially improve mood, both smiling and laughing did. Moreover, laughter seemed to boost positive affect more than just smiling by 22 adults. PMID- 12585517 TI - Confirmatory factor analysis and reliability of the Chinese version of the Maslach Burnout Inventory among guidance teachers in Hong Kong. AB - In 1995 Chan and Hui examined the responses of a sample of Chinese teachers on the Maslach Burnout Inventory and recommended a possible 2-factor rather than the original 3-factor model for the assessment of burnout among Chinese teachers. In the present study, the factor structure of responses to the Chinese version of the Maslach Burnout Inventory in a sample of 1,398 Chinese secondary school guidance teachers was examined using the EQS approach to confirmatory factor analysis. The results showed that a 3-factor model (Emotional Exhaustion, Depersonalization, and Personal Accomplishment) provided the best fit, with the first two factors highly correlated (r = .80). Internal consistencies for the subscales ranged from .80 to .88. PMID- 12585518 TI - What proportion of heterosexuals is ex-homosexual? AB - How many heterosexuals are 'ex-homosexuals'? In 1984, a random sample of Dallas adults indicated that 8 (2.7%) of 294 currently heterosexual men and 4 (1.0%) of 393 currently heterosexual women said that they were ex-homosexual. Of an urban sample from 5 additional cities, 0.5% of current heterosexuals reported that they had been homosexually 'married'. It thus appears that perhaps 1-2% of heterosexuals are ex-homosexuals. Proportionately more adults than teenagers and more men than women moved from homosexuality to heterosexuality. Of the 18 who changed, 12 became heterosexual and 6 bisexual, suggesting that perhaps two thirds of those who abandon "being" homosexual 'become' heterosexual and a third 'become' bisexual. Because labeling oneself 'homosexual' is so mutable and value laden, the term 'omnisexual' is suggested. PMID- 12585519 TI - Psychometric properties of Spence and Robbins' measures of workaholism components. AB - There has been a recent increase in attention devoted to the study of workaholism. As with most new areas of study, issues of definition and measurement have not received their due. The present investigation examined some psychometric properties of Spence and Robbins' measures of the components of workaholism (1992), one of two measures that have received some attention. These measures were relatively stable in a sample of senior Norwegian managers (N = 87) over a 6-mo. period, had generally acceptable internal consistency reliability, and showed factor structures supportive of three components of workaholism. PMID- 12585520 TI - Government policies, industry/economics, social trends, and educational opportunities in 'women's decisions to work outside versus inside the home. AB - Research on the career decision processes of women has focused primarily on internal considerations such as mathematical ability, intelligence, and self esteem; however, the external environment also has an influence on these processes. To date, the environmental factors have primarily been researched separately, if at all. They include government policies, industry/economics, social trends/expected sex roles, and educational opportunities. This pilot study investigated these four external factors simultaneously. In addition, a survey of literature on career decision indicated such research to be based on a limited pool of highly educated, management/professional women. This study also uses a wider group including housewives, blue collar, and pink collar women. PMID- 12585521 TI - Relation of managers' personality to accident and injury rates. AB - Previous research has explored the link between certain aspects of individuals' personalities and their safety behavior and accident rates. We predicted a relation between 23 managers' scores on Big Five personality dimensions and departmental accident and injury rates in a manufacturing company. Accident and injury data were provided by the organization. We found a relation between managers' personality profiles and both injury and accident rates in their departments. The findings suggest that managers with certain types of personalities may have safer work units. This has implications for the selection of managers in work environments with safety risks. PMID- 12585522 TI - Psychometric properties of the Spanish form of Templer's Death Anxiety Scale. AB - The aim of this study was to translate and adapt Templer's Death Anxiety Scale into Spanish and examine mean scores, internal consistency, temporal stability, and factor structure for a sample of 187 Spanish students, 147 women and 40 men, of an average age of 20.8 yr. Mean scores were 6.2 (SD = 3.4) for women and 5.0 (SD = 2.7) for men; these differences were statistically significant. Cronbach coefficient alpha was .73, and test-retest correlation over 3 wk. was .87. Analysis by principal components with varimax rotation produced four factors with eigenvalues greater than one; these were labeled (1) Cognitive-Affective, (2) Pain and Illness, (3) Stimuli Related to Death, and (4) Awareness of Time Passing. Obtained results were consistent with previous studies carried out on other versions of the scale and indicate this Spanish form of the Death Anxiety Scale is useful for evaluating death anxiety in Spanish-speaking subjects in Spain. PMID- 12585523 TI - Psychometric assessment of measures of psychological symptoms, functional status, life events, and context for low income Hispanic patients in a primary care setting. AB - This study assessed the psychometric properties of a set of nine commonly used lists in an English-speaking Hispanic sample taken from a primary care setting. Those were the General Health Questionnaire-12, the Anxiety, Depression, and Somatization scales from the SCL-90, the Medical Outcomes Study Short Form-36, the Holmes and Rahe Family Life Changes, abbreviated versions of the Daily Hassles and Uplifts, the Herth Hope Index, Duke Social Support and Stress scales, Alcohol Use Disorders Identification Test, and Marlowe-Crowne Brief Social Desirability Scale. 68 adult patients completed the measures via structured interview. Internal consistency was measured using Cronbach alpha and the Kuder Richardson-20. Construct validity was assessed using Pearson correlations among sets of scores. Internal consistencies were good-to-excellent for all measures except for the Duke Social Support and Stress scales (Stress scale), and the Brief Social Desirability Scale. Construct validity was suggested for all except the Mental Health and Energy/Fatigue scales of the Short Form-36. Further study is needed to verify these results in other Hispanic populations and to address other forms of reliability and validity. PMID- 12585524 TI - Effectiveness of the Responsible Social Values Program for 6th grade students in one rural school district. AB - The present study reports the effectiveness of an abstinence-based program (the Responsible Social Values Program) designed for 6th grade students in a rural school district. To assess whether this 6-hr. abstinence-based program changed students' knowledge of their sexual development during adolescence, attitudes toward early sexual involvement, intentions regarding sexual involvement, views related to expected consequences of being sexually involved, and views related to expected outcomes of being sexually involved, a pre-experimental one-group, pretest-posttest design was employed. Analysis indicated that the curriculum was effective in terms of improving scores for each of the constructs measured. Immediately after the 6-hr. program, more students stated that they planned to avoid having sex until marriage (70%) than prior to the program (50%). Replication with a control group and longer term follow-up testing is necessary. PMID- 12585525 TI - Meta-analysis of relations of stress propensity with subjective stress and strain. AB - Meta-analyses were used to examine relations of three factors of stress propensity, i.e., cognitive-affective connectivity, psychological magnification, and anger-irritability, with three stress criteria, i.e., subjective stress, psychological strain, and physiological strain. Each of the stress propensity factors was significantly related to each criterion. Sex was a significant moderator for two of these nine relations (cognitive-affective connectivity with subjective stress and anger-irritability with psychological strain). PMID- 12585526 TI - Women's advantage on verbal memory is not restricted to concrete words. AB - It is well established that women perform better than men on tests of verbal memory, but the nature of this advantage is unclear. To examine whether reference to a real object is a factor, we presented several verbal memory tasks, including one containing words high and low in concreteness. Over all tests there was an expected female advantage. Although concrete words were recalled much better than abstract words, the female advantage was equivalent on both. The sex difference was not accounted for by a difference in verbal intelligence. Possible brain and adaptive mechanisms are discussed. PMID- 12585527 TI - The Chinese version of the Marital Comparison Level Index revisited. AB - An attempt was made to tailor the Chinese version of the Marital Comparison Level Index to the Hong Kong Chinese context by adding 11 items. Analysis showed that the amended version is a reasonably reliable and culture-appropriate measure for research. PMID- 12585528 TI - Responses to lost letters about a 2000 General Election amendment to abolish prohibition of interracial marriages in Alabama. AB - A field study using 621 "lost" letters was conducted in the city of Mobile and in small towns in mostly rural Baldwin County, Alabama. Milgram's lost letter technique was validated against the actual votes cast during the November 7, 2000 General Election. The technique was successful as an unobtrusive measure useful for predicting patterns of voting behavior. Rates of return of lost letters "in favor of and opposed to legalizing interracial marriage" agreed with the actual election returns (chi-square "goodness of fit"). Community size seemed associated with return of lost letters. PMID- 12585529 TI - Adjectives, phrases, and sentences: intracorrelation of three five-factor personality inventories. AB - The Eysenck Personality Inventory and three short five-factor personality inventories, the NEO Five Factor Inventory, the Berkeley Personality Profile, and the SONSO Personality Inventory, were administered in pairs to six samples of college subjects. The three samples using the Eysenck inventory each contained 300 subjects, whereas the remaining three samples contained 500 subjects each. Each sample was half women and half men. The inventories appear to have corresponding scales, often with correlations of around .70 among them. Also, measures associated with Neuroticism were weakly and negatively correlated with some measures of Extraversion, Agreeableness, and Conscientiousness. PMID- 12585530 TI - Individualism and collectivism: cultural orientation in locus of control and moral attribution under conditions of social change. AB - This study examined the validity of the view that the constructs of individualism and collectivism are coherent cultural manifestations necessarily reflected in an individual's attribution patterns. It was hypothesized that the attribution patterns of locus of control and moral accountability would show divergent individualistic and collectivistic influences in a culture during change from a collectivist culture to an individualist culture. 98 university students from the United States and Puerto Rico were administered the Singelis Individualism Collectivism Scale, Rotter's Locus of Control Scale, and Miller and Luthar's justice-related moral accountability vignettes. Contrary to expectation, the Puerto Rican sample scored less external in locus of control than the United States sample. No cultural differences in moral accountability were found. No strong correlations were found among the variables at the individual level of analysis. Accounting for these results included the lack of representativeness of the samples, the independence of relation between variables at different levels of analysis, and social change. PMID- 12585531 TI - Evaluating the relation between memory and intelligence in children with learning disabilities. AB - Although both intelligence tests and memory tests are commonly used in neuropsychological examinations, the relationship between memory and intelligence has not been fully explored, particularly for children having learning disabilities. Memory, or the ability to retain information, was evaluated using the Test of Memory and Learning, a recently released test that gives a comprehensive measure of global memory functioning. This, and the Wechsler Intelligence Scale for Children-Third Edition, used to assess intelligence, were given to 80 students with learning disabilities. The correlation between a global measure of memory and a global measure f intelligence was significant (r = .59), indicating that memory should be viewed as an important component when evaluating children with learning disabilities. PMID- 12585532 TI - Survey return rates using a covering letter signed by a graduate student or faculty member. AB - This study assessed differences in response rate when the author of a survey was identified as a graduate student or a faculty member. A survey on research ethics was mailed to 500 nurse faculty members with half of the covering letters signed as a graduate student and half signed as a faculty member. There were 388 (78%) out of a possible 495 surveys returned. The response rate was 78% for graduate student and 77% for faculty identifications, not significant by chi-square test. PMID- 12585533 TI - Attachment to the physical dimension of places. AB - Social relationships had been important in explanation and prediction of attachment to places. Although some have asserted the importance of physical aspects of the environment in the formation of attachment ties to a place, the social environment is required for the formation of bonds to a place, although strong emphasis on the social aspect has been questioned and the importance of the physical environment noted. The present objective in two studies was to test whether college students (ns = 30 and 27) show a preference for a place they know, independently of the social interactions developed in them. Results confirmed the hypothesis, i.e., after a very brief stay in a certain place with nobody else there, these college students preferred that place to another with which they had not had previous contact. PMID- 12585534 TI - Ratings of characteristics of ethnic groups by authoritarians and nonauthoritarians when stereotypes vary in favorability. AB - White non-Jewish participants in the 1990 General Social Survey with low scores on Tolerance of deviation ("Intolerant"), a measure of authoritarianism, rated African Americans more unfavorably than did Tolerant respondents on two traits, intelligence and laziness, when stereotypes about the latter group are unfavorable. But the Intolerant were not more negative in their evaluations than the Tolerant when Jews, a group for whom the stereotypes on these traits are favorable, were rated. The Intolerant rated Jews more favorably on intelligence, and their ratings of laziness did not differ from those by the Tolerant. PMID- 12585535 TI - Job-related motivational factors among Malaysian employees. AB - This study identified job-related motivational factors among Malaysian employees in several telecommunication companies. Responses were obtained from 1,179 employees at all levels up to senior managers and six different functional divisions, sales and marketing, human resources, finance, technical, information, technology, and support division. All employees were asked to rate the importance of Kovach's 10 job-motivational factors. These factors were good wages, job security, opportunity for career growth in the organization, good working conditions, interesting work, company loyalty to employees, tactful discipline, full appreciation of work done, sympathetic help with personal problems, and feeling of being involved in the organization. The top five factors employees identified as motivating them in their jobs were good wages, job security, company loyalty to employees, good working conditions, and full appreciation for work done. Findings were in accordance with Kovach for U.S. employees, in which the top motivational factors were good wages and job security. PMID- 12585536 TI - Cult suicide and physician-assisted suicide. AB - A greater proportion of Dr. Kevorkian's physician-assisted suicides and the Heaven's Gate cult suicides appear to be women than the general population of suicides. PMID- 12585537 TI - Comparison of two exercise programs on general well-being of college students. AB - Responses to life stressors are associated with negative behaviors that may increase risk for illness and injury. The effect of high intensity exercise in reducing reactivity to psychological stress has been well documented among older people. The purpose of this study was to ascertain the effect of weight-training versus aerobic dance on psychological stress in college students. 45 students participated in a weight-training course, 35 students participated in aerobic dance classes, and 34 students served as a control group. The Survey of Recent Life Experiences was used to appraise stressfulness of current experiences before and after exercise intervention. On immediate retest after 8 wk. of weight training perceived stress was significantly reduced when compared with an 8-wk. aerobic dance program, but there were no significant differences between the control group and the weight-training group or the aerobic dance group. These results suggest that a regular routine of low intensity exercise such as weight training may reduce perceived stress on an immediate test. PMID- 12585538 TI - Correlations of sense of humor and sleep disturbance ascribed to worry. AB - Scores on the Sleep Disturbance Ascribed to Worry Scale were correlated with total scores on the Multidimensional Sense of Humor Scale and subscale sleep scores. A significant negative correlation of -.22 (p < .01) was found between the disturbance scores and scores on the humor production subscale. Research is required to replicate and extend these findings. PMID- 12585539 TI - Role of education in generativity differences of employed and unemployed women in Korea. AB - This study investigated generativity differences between employed and unemployed women in Korea and examined the effect of education on generativity. There were 472 participants in this study, 252 employed married women and 220 unemployed married women living in the Kwangju metropolitan area. A questionnaire requesting demographic information and responses to the translated Loyola Generativity Scale was administered individually. Analysis showed significant generativity differences between the two groups, who also differed in education and mother and spouse roles. However, employment status was not a significant predictor for generativity when a stepwise regression analysis was applied. The analysis showed that education was the strongest predictor for generativity, while mother/wife roles, socioeconomic status, health, and childcare stress were also significant predictors. It was concluded that for Korean married women, generativity is more strongly related to education than employment status. PMID- 12585540 TI - Figure-drawing indices of psychological accessibility. AB - We investigated the hypotheses that windows, doors, pathways, and size of the house in the House drawing of the House-Tree-Person projective drawing technique reflect psychological and social accessibility by correlating these variables with the Social Introversion scale of the MMPI-2. Subjects were 153 nonclinical, community adults, and 146 clinical patients currently receiving mental health services. While there were strong correlations among these figure-drawing variables, suggesting some common underlying dimension, these scores were not significantly correlated with psychological accessibility as measured by the Si scale of the MMPI-2. Possible interpretations were suggested. PMID- 12585541 TI - Buying textbooks online. AB - In a sample of 77 undergraduates, purchasing textbooks online was predicted by computer/Internet skills but not by personality traits. PMID- 12585542 TI - Data-dependent analyses in psychological research. AB - A data-dependent analysis assumes that all the information available to a researcher is contained within the observed data. Data-dependent methods for the analysis of experimental designs are shown to provide significant advantages over conventional techniques such as an F test. Two versions of three data-dependent methods based on permutations of the data are described and compared. One version utilizes ordinary least squares regression, and the other version utilizes least absolute deviations regression to analyze experimental designs. Analyses of an unbalanced two-way experimental design illustrate the differences among the six data-dependent approaches and the classical ordinary least squares F test, which depends on the assumptions of normality, homogeneity, and independence. PMID- 12585543 TI - Reliability of coordinate grid data. AB - The reliability of data obtained from a modified repertory grid procedure was examined. The coordinate grid, developed by Chambers (1983), measures the logical inconsistency and integrative complexity of an individual's view of different people. 56 college students completed two forms of the coordinate grid on each of two testing occasions. Analysis indicated low reliability for measures of both logical inconsistency (r = -.04) and integrative complexity (r = .31) with respect to the people entered into the grid. The test-retest results were better (r = .51 for logical inconsistency; r = .35 for integrative complexity) but still well below acceptable reliability. Hence, generalizations beyond a particular grid may be unwarranted. PMID- 12585544 TI - Castration anxiety and phobias. AB - Based on Freud's case study of "Little Hans," the authors tested the hypothesis that men with phobias would score higher on castration anxiety than men without phobias. College men with either average or high scores on the Fears Scale of the MMPI-2 (n = 10 men in each group) responded to the Thematic Apperception Test, which was scored for castration anxiety. Men with high scores on the Fears Scale had higher scores on castration anxiety than men with average scores on the Fears Scale. The findings are consistent with Freud's hypothesis about phobias. PMID- 12585545 TI - Effects of leaders' and evaluators' sex on sex-role stereotyping of charismatic leaders. AB - This study extends research on sex and leadership by examining the relation between evaluators' and leaders' sex and the sex-role stereotyping of charismatic leaders. A total of 219 students (110 men and 109 women) from a large northeastern university rated charismatic leaders depicted in vignettes using the revised Bem Sex-Role Inventory: overall, the leaders were rated higher on masculinity than femininity. Analysis by sex of evaluator and leader showed masculinity was higher in all cases except when male charismatic leaders were evaluated by women. In this case, the results support an androgynous view, i.e., high on both masculinity and femininity. PMID- 12585546 TI - Essentialist thinking about depression: evidence for polarized beliefs. AB - Mixture modeling of 309 undergraduates' beliefs about the nature of depression yielded two polarized latent classes--those who held essentialist beliefs and those who did not--consistent with the view that essentialist thinking is a distinct cognitive mode. PMID- 12585547 TI - An evaluation of the Organizational Learning Survey. AB - A mail survey of a national random sample of 2,000 marketing managers was conducted. The data provided by 221 respondents were analyzed to assess the unidimensionaliry of the 21-item 1997 Organizational Learning Survey developed by Goh and Richards. A confirmatory factor analysis using LISREL 8.30 did not support the unidimensionality of the 21-item survey; however, unidimensionality was established for most of the dimensions. Managerial implications and directions for research were suggested. PMID- 12585548 TI - Comparison of medical students, medical school faculty, primary care physicians, and the general population on attitudes toward psychological help-seeking. AB - This study is a preliminary comparison of the attitudes of osteopathic medical students, medical school faculty, primary care providers, and the general population toward seeking professional psychological help. Attitudes were also studied in the former three groups for those who had and had not previously received mental health services. 103 medical students, 22 faculty, 31 primary care providers, and 395 people from the general population responded to the mail out survey. Attitudes toward help-seeking were more negative among the general population group than among students and providers. For these students, faculty, and providers, attitudes toward seeking help were more positive if they reported having received mental health services in the past. PMID- 12585549 TI - Mathematics self-concept and mathematics anxiety of undergraduate majors in education. AB - This study investigated the relation between mathematics self-concept and mathematics anxiety of a sample of undergraduate majors in education. Scores of 68 students (10 men and 58 women) on mathematics self-concept were positively correlated (.83) with mathematics anxiety. PMID- 12585550 TI - Sibling effects, environmental influences, and school dropout. AB - Relations were examined between sibling variables, environmental influences, and the likelihood of dropping out of school. Data were collected from Australian adolescents (4,096 boys, 4,570 girls). Findings indicated that, after taking into account measures of distal contexts and proximal learning settings, sibling variables continued to have small but significant associations with adolescents' school dropout behavior. PMID- 12585551 TI - Solving nonproblems. PMID- 12585552 TI - Oversight. PMID- 12585553 TI - Targeting obesity. PMID- 12585554 TI - The hazy hormone landscape. PMID- 12585555 TI - Pinning hopes on hormones. PMID- 12585556 TI - Endocrine disrupters as a public health issue. PMID- 12585557 TI - Hormone therapy in menopause: the plot thickens. AB - The termination of one of the trials in the Women's Health Initiative (WHI) study is causing turmoil in women using hormone therapy. Practitioners and patients alike are confronted with conflicting evidence about the safety and effectiveness of both estrogen therapy (ET) and hormone (estrogen/progestin) therapy (HT). Recent investigations of the estrogen receptor, the genome, and cellular cofactors have demonstrated the heterogeneity of molecular response to hormones. This implies a heterogeneous response in the individual as well and casts doubt on the validity of prior research. Recent experiments in monkeys have provided a possible explanation for the adverse effects of HT on the cardiovascular system that were detailed by the Heart and Estrogen/Progestin Replacement Study (HERS) and WHI studies. This article explores nuances of several key studies and discusses potential therapeutic options, including use of lower-dose estrogen, transdermal estrogen, selective estrogen-receptor modulators, and diminished use of progestin. PMID- 12585558 TI - Osteoporosis: alternatives to estrogen treatment. AB - Osteoporosis-related fractures result in significant morbidity, mortality, and cost in the United States. With the aging of our population, the cost to society and number of individuals who will become disabled by hip fractures alone could triple by the year 2040. Many known risk factors allow us to identify individuals with increased susceptibility for osteoporosis. Secondary causes of bone loss should always rigorously be ruled out in patients. Laboratory assessment and measurement of bone mineral density can help aid treatment choices. This article discusses risk factors for and secondary causes of osteoporosis, as well as radiographic and laboratory evaluation for the disease. It also presents nonestrogen treatments for osteoporosis. Lifestyle changes are an important aspect of treatment. In addition, antiresorptive and anabolic agents can markedly improve bone mineral density. Our ability to prevent fractures will be improved in the future as we are better able to assess fracture risk, measure bone strength, and provide new compounds for the prevention and treatment of bone loss. PMID- 12585559 TI - Vitamin D--the steroid hormone prescription for every patient. AB - Physicians in the United States rarely screen for hypovitaminosis D and rarely prescribe vitamin D, even when medically indicated. This is of particular concern in Minnesota. The sun's intensity at Minnesota's latitudes limits vitamin D production, at best, to March through October. A variety of lifestyle situations, including long work hours, may preclude adequate sun exposure. Additionally, people of Northern European background may avoid sun exposure to reduce risk of skin cancer and premature aging. And people of Asian and African heritage may not have sufficient vitamin D production due to increased skin pigmentation. This brief article summarizes key points regarding the importance of vitamin D, including its action as a steroid hormone and its role in cancer, hypertension, and autoimmune disease as well as in perinatal and prenatal health. The potential benefit of hypovitaminosis D screening and vitamin D supplementation is discussed, as are the populations most likely to need screening and supplementation. PMID- 12585560 TI - Hypertension, obesity, and the renin-angiotensin system: a tale of tight associations. AB - Obesity, hypertension, and their co-morbid conditions such as diabetes mellitus and cardiovascular disease are major public health issues. This article reviews research that links hypertension and obesity, particularly the android obesity pattern that characterizes the metabolic syndrome. The article focuses on the relationships between hypertension, obesity, and the renin-angiotensin system (RAS). It explores new understandings about the RAS and the connections between RAS overactivity, hypertension, and obesity. Also discussed are lines of evidence that show that elevated visceral fat levels contribute to renin-angiotensin overactivity and that RAS overactivity contributes to the development of obesity. The bidirectional nature of the obesity-RAS relationship is discussed. PMID- 12585561 TI - And the suit goes on: reporting malpractice settlements and verdicts to the board of medical practice. PMID- 12585562 TI - The great dictator. PMID- 12585563 TI - The pharmacologic management of SSRI-induced side effects: a survey of psychiatrists. AB - Despite the superior side effect profile of the newer antidepressants over the tricyclics and monoamine oxidase inhibitors, all newer antidepressants are associated with a wide array of side effects. Clinicians are constantly confronted with the challenge of managing these side effects in the context of minimal research to prove one management strategy is more effective than another. The purpose of this study was to examine prescribing practices regarding the management of SSRI-associated side effects in a sample of psychiatrists attending a psychopharmacology review course. A total of 439 out of 800 clinicians (55%) attending a psychopharmacology review course responded to our questionnaire that was given prior to beginning the review course, though not all respondents answered all four items on the questionnaire. Among these items were questions designed to assess clinician preference for the management of SSRI-induced side effects. As a treatment for SSRI-induced sexual dysfunction, 43% (143/330) chose adding bupropion, while 36% (120/330) opted to switch agents as their first choice; for SSRI-induced insomnia, 78% (264/326) chose adding trazodone. Switching agents was the first choice of 61% (214/353) of clinicians for managing SSRI-induced agitation, 93% (339/363) for managing SSRI-associated weight gain. In an effort to manage most SSRI-associated side effects (with the exception of sexual dysfunction and insomnia), the majority of the clinicians responding to our survey opted to switch agents rather than add a specific medication to the existing SSRI. In our opinion, this practice may reflect the relative lack of research studies on the role of adjunctive treatments in the management of SSRI induced side effects and/or the tendency to favor monotherapy over polypharmacy. PMID- 12585564 TI - Cigarette smoking among adolescent psychiatric inpatients: prevalence and correlates. AB - The aim of this study was to ascertain the rate of smoking among adolescent (13 17 years old) psychiatric inpatients and to examine factors related to smoking status in this population. The medical records of all adolescents admitted to a private psychiatric hospital during a 1-year period were reviewed. The rate of current smoking was 59.8%. Of the adolescents who reported smoking, 40% smoked at least 140 cigarettes per week, and 44.7% smoked 7 days a week. Smokers were more likely to be Caucasian, fall within the 13-year age group, and meet criteria for substance abuse/dependence. Adolescents who used no alcohol or illicit substances were least likely to report current smoking, followed by those who used alcohol only, followed by those who used illicit substances. Results of the current study suggest the need for both thorough assessment of smoking and specialized smoking interventions in this population, who are likely to mature into heavily dependent, recalcitrant adult smokers in the absence of intervention. PMID- 12585565 TI - Effectiveness of pharmacotherapy for pathological gambling: a chart review. AB - Although pathological gambling is a relatively common disorder, there exists only limited information regarding the effectiveness of pharmacotherapy for this illness. This study examines which medications may be effective, dose and duration of medication trials needed to achieve response, and possible predictors of response. Using a chart review, 50 adult outpatients with DSM-IV pathological gambling treated in clinical practice were assessed regarding response to a variety of medications, including augmentation strategies, and response to concomitant psychotherapy. All subjects received pharmacotherapy for gambling symptoms. Thirty-nine (78%) achieved response to medication treatment. Mean duration of treatment needed for response was 104.9 +/- 85.0 days. Of those treated with an adequate trial of naltrexone as monotherapy, 90.9% were responders, whereas only 45.5% of those treated with an adequate trial of an SSRI achieved response. Patients with poorer social and occupational functioning due to urges and thoughts about gambling were less likely to respond to medication. These findings from a clinical setting suggest that a majority of pathological gamblers improve with medication treatment. Naltrexone, or augmentation of naltrexone with an SSRI, appears to be most effective in relieving gambling symptoms. PMID- 12585566 TI - The effects of atypical antipsychotics on serum prolactin levels. AB - Hyperprolactinemia may be a concern in the treatment of patients with schizophrenia. The side effects associated with high prolactin levels can have a negative impact on patient compliance with treatment. Atypical antipsychotics as a group cause less hyperprolactinemia than conventional antipsychotics, yet there is considerable variation among specific drugs. Risperidone at higher doses has been shown to produce increases in prolactin similar to conventional antipsychotics. At the other end of the spectrum, clozapine and quetiapine produce minimal sustained increases in prolactin that are no different from placebo. However, correlations between prolactin elevations and clinical symptoms have not been well-established. This paper reviews the published literature regarding prolactin levels in treated and untreated patients with schizophrenia and the relationship of prolactin and dopamine. It concludes with an overview of the effects of specific atypical antipsychotics on prolactin levels in patients with schizophrenia. PMID- 12585567 TI - Long-term side effects of newer-generation antidepressants: SSRIS, venlafaxine, nefazodone, bupropion, and mirtazapine. AB - Anecdotal reports have suggested that the long-term use of selective serotonin reuptake inhibitors (SSRIs) may be associated with significant weight gain, sexual dysfunction, drug interactions, and discontinuation symptoms. Are these effects inevitable or can they be managed effectively with the appropriate interventions? In reviewing published, controlled clinical trials, it has been noted that many depressed patients experience weight gain during remission with or without treatment. Most antidepressants appear to produce a 3- to 4-kg weight gain after 6-12 months of therapy, which may be managed with nutritional counseling and exercise. The exception is mirtazapine, which appears to be associated with significant weight gain early in therapy. Antidepressant-induced sexual dysfunction is also common but may be managed with the addition of an antidote or substitution. Drug interactions are most common with fluvoxamine, nefazodone, and fluoxetine because these agents are more likely to affect the metabolism of commonly prescribed medications. It may be possible to prevent discontinuation symptoms with a cross taper to another antidepressant or by slowly tapering the antidepressant. PMID- 12585568 TI - Sleep in posttraumatic stress disorder. AB - Posttraumatic stress disorder (PTSD) is often associated with sleep disturbances. In this review, we focus on the published literature on subjective and objective findings of sleep in patients with PTSD. Insomnia and nightmares are most commonly reported subjective sleep disturbances. Polysomnographic investigations have frequently reported rapid eye movement (REM) sleep abnormalities in PTSD. However, studies have not been consistent about the type of REM sleep dysfunction in PTSD patients. Antidepressants such as nefazodone, trazodone, fluvoxamine, and imagery rehearsal therapy are found to be beneficial in the treatment of PTSD associated sleep disturbances as well as core symptoms of this anxiety disorder. We propose use of such modalities of treatment in PTSD patients with predominant sleep disturbances. Further studies are required to clarify polysomnographic sleep changes especially role of REM sleep dysregulation and treatment of sleep disturbances in PTSD. PMID- 12585569 TI - The National Joint Registry for England and Wales. PMID- 12585570 TI - The management of partial-thickness tears of the rotator cuff. PMID- 12585571 TI - Complications of arthroscopic reconstruction of the anterior cruciate ligament. PMID- 12585572 TI - The 'French paradox.'. PMID- 12585573 TI - Surgical reconstruction of late pelvic post-traumatic nonunion and malalignment. AB - We have retrospectively reviewed the clinical and radiological results in 204 consecutive adult patients who had surgical correction of 70 late post-traumatic pelvic nonunions and 134 malalignments. The deformed pelvises were subdivided into united (true), unstable, ununited, and partially stable malalignments with heterotopic bone. The principal complaints were of pain, pelvic instability, sitting imbalance, and apparent limb-length discrepancy. After surgery, 195 patients (96%) achieved a primary union and 144 (71%) had slight, intermittent or no pelvic pain, while pelvic instability was entirely eliminated. Overall, 131 patients (64.2%) were extremely satisfied, 58 (28.4%) were satisfied and 15 (7.4%) were unsatisfied. After reconstruction of the malaligned pelvises, 67 results (50%) were anatomical, 47 (35%) were satisfactory and 20 (15%) were unsatisfactory. For a pelvic nonunion with local osteopenia and malalignment, stabilisation of all three pelvic columns is recommended. True pelvic (united) malunions were the most satisfactorily realigned and had the fewest complications. Ununited and unstable malalignments, especially those with heterotopic bone, had the poorest corrections and the most neurological complications. A therapeutic alternative, by the local resection of a symptomatic bony prominence, and fixation in situ of a posterior pelvic nonunion, gives highly effective symptomatic relief with fewer complications. Despite this, many patients had persistent low back pain. PMID- 12585574 TI - The relationship between cup design and the radiological signs of aseptic loosening in total hip arthroplasty. AB - We assessed differences in the incidence and appearance of the radiological signs of loosening of the cup for various types of design. This was an observational study based on hip registry data of 15,340 patients with 17,951 total hip arthroplasties collected over a period of 33 years in 49 hospitals in Central Europe. The threaded and the press-fit titanium cups showed significantly less aseptic loosening than the other systems. The direction of migration and the frequency of the radiological signs of loosening differed between the cup systems and were time-dependent. Our findings indicate the superiority of uncemented threaded cups and press-fit titanium cups over other designs of cup. PMID- 12585575 TI - The validity of clinical examination in the diagnosis of loosening of components in total hip arthroplasty. AB - We analysed follow-up data from 18,486 primary total hip arthroplasties performed between 1967 and 2001 to assess the validity of clinical procedures in diagnosing loosening of prosthetic components. Sensitivity, specificity and predictive values were estimated with the radiological definition of loose or not loose as the 'gold standard'. The prevalence of acetabular loosening increased from 0.6% to 13.9% during the period of the study and that of femoral loosening from 0.9% to 12.1%. Sensitivities and positive predictive values were low, suggesting that clinical procedures could not replace radiological assessment in the identification of loose prostheses. Specificities and negative predictive values were constantly above 0.86. The possibility of there being a prosthesis which is not loose in asymptomatic patients was consequently very high, particularly during the first five to six years after operation. The necessity of periodic clinical and radiological follow-up examinations of asymptomatic patients during the first five to six years after operation remains questionable. Symptomatic patients, however, require radiological assessment. PMID- 12585576 TI - Should the cement mantle around the femoral component be thick or thin? AB - We have compared the survival and radiological outcome at ten years after total hip replacement using two techniques for preparing the femoral canal. The same prosthesis was used throughout and all operations were performed by the same surgical team. In technique 1 the canal was over-reamed by 2 mm and in technique 2 it was reamed to the same size as the prosthesis. Technique 1 was performed on 92 patients and technique 2 on 97 patients. The survival at ten years was 97.2% (90.6 to 99.2) for technique 1 and 98.8% (92.9 to 99.8) for technique 2. Vertical migration was greater in technique 1 (1.8 mm versus 1.0 mm at five years; p = 0.36). There were significantly more lytic lesions and radiolucent lines at five years (p = 0.0061) with technique 1. We conclude that technique 2 is not worse and may produce better long-term results than current teaching suggests. PMID- 12585577 TI - Patient demographics as a predictor of the ten-year survival rate in primary total knee replacement. AB - s the surgical indications for total knee replacement (TKR) expand to include younger, heavier and more active patients, knowledge of the effect of these demographic variables on the outcome and survival of the implant is increasingly important. Between November 1986 and September 1990, 402 patients underwent 562 primary cemented cruciate-retaining TKRs carried out by a single surgical team. The overall results showed a survival of 96.8% at 14 years with 1.44% lost to follow-up. Evaluating the demographics of these patients showed that certain groups fared significantly less well. The best results were seen in non-obese women with osteoarthritis who were over 60 years of age in whom there was ten year survival of 99.4%. The worst results were in obese men with osteoarthritis who were less than 60 years of age in whom there was a ten-year survival of 35.7%. Caution should be exercised when considering TKR on a patient with this combination of poor risk factors. By identifying demographic factors at the time of consultation the surgeon is better able to predict the survival of the TKR. This information is important when considering the best options for treatment of a patient and in providing accurate information during preoperative counselling. PMID- 12585578 TI - Arthrodesis of the knee using a custom-made intramedullary coupled device. AB - Nine patients underwent arthrodesis of the knee using customised coupled nail (the Mayday arthrodesis nail), five after infected arthroplasty, one following failed arthrodesis, one for intractable anterior knee pain, one for Charcot instability and one after trauma. Comparison was made with 17 arthrodeses, eight undertaken using external fixation, four with dual compression plates, and five with long Kutntscher nails. Union was achieved in all patients (100%) at a mean time of ten months using the customised implant. There were no complications despite early weight-bearing. No further procedures were required. This contrasted with a rate of union of 53% and a complication rate of 76% with alternative techniques. Of this second group, 76% required a further operative procedure. We compared the Mayday arthrodesis nail with other techniques of arthrodesis of the knee. The differences in the need for further surgery and occurrence of complications were statistically significant (p < 0.001), and differences in the rate of nonunion and inpatient stay of less than three weeks were also significant (p < 0.05) using Fisher's exact test. We conclude that a customised coupled intramedullary nail can give excellent stability allowing early weight-bearing, and results in a high rate of union with minimal postoperative complications. PMID- 12585579 TI - A mobile-bearing total knee prosthesis compared with a fixed-bearing prosthesis. A multicentre single-blind randomised controlled trial. AB - Before proceeding to longer-term studies, we have studied the early clinical results of a new mobile-bearing total knee prosthesis in comparison with an established fixed-bearing device. Patients requiring bilateral knee replacement consented to have their operations under one anaesthetic using one of each prosthesis. They also agreed to accept the random choice of knee (right or left) and to remain ignorant as to which side had which implant. Outcomes were measured using the American Knee Society Score (AKSS), the Oxford Knee Score (OKS), and determination of the range of movement and pain scores before and at one year after operation. Preoperatively, there was no systematic difference between the right and left knees. One patient died in the perioperative period and one mobile bearing prosthesis required early revision for dislocation of the meniscal component. At one year the mean AKSS, OKS and pain scores for the new device were slightly better (p < 0.025) than those for the fixed-bearing device. There was no difference in the range of movement. We believe that this is the first controlled, blinded trial to compare early function of a new knee prosthesis with that of a standard implant. It demonstrates a small but significant clinical advantage for the mobile-bearing design. PMID- 12585580 TI - Minimal internal fixation and calcium-phosphate cement in the treatment of fractures of the tibial plateau. A pilot study. AB - We used calcium-phosphate cement combined with minimal internal fixation to treat 49 fractures of the lateral tibial plateau. There were 25 split depression fractures, 22 pure depression fractures and two bicondylar fractures. Anatomical reduction was obtained in 38 fractures, satisfactory reduction in nine and imperfect reduction in two. Of 44 patients reviewed at one year, 33 were rated as having an excellent reduction. Functional outcome as measured by the Rasmussen score was good or excellent at six months in 92% (44/48) of patients and in 95% (42/44) at one year. Eight (16%) showed some loss of reduction of the plateau. In seven of these the loss of reduction was slight (<3 mm) and no action was taken. One patient with a deep infection had gross loss of reduction and a poor functional outcome. Calcium-phosphate cement is a useful alternative to bone grafting for the treatment of fractures of the tibial plateau. PMID- 12585581 TI - Use of the coventry infant hip screw in the treatment of nonunion of fractures of the distal humerus. AB - Between 1993 and 2000 we treated 14 patients with nonunion of fractures of the distal humerus; 11 had already had previous fixation which had failed. The mean time to surgery was 21 months after injury. All the fractures were very low which made it difficult to obtain a firm hold of the small distal fragment. The problem of fixation was addressed by inserting a Coventry infant hip screw into the humeral condyles. The screw has a thread of wide diameter which gives excellent purchase on the small distal fragment. The condyles can then be compressed on to the humeral shaft using a 4.5 mm narrow tibial dynamic plate. Of these 14 difficult cases of nonunion, 12 progressed to union. PMID- 12585582 TI - Aberdeen Colles' fracture brace as a treatment for Colles' fracture. A multicentre, prospective, randomised, controlled trial. AB - We carried out a randomised, prospective, multicentre clinical trial of the treatment of Colles' fractures. A total of 339 patients was placed into two groups, those with minimally displaced fractures not requiring manipulation (151 patients) and those with displaced fractures which needed manipulation (188 patients). Treatment was by either a conventional Colles' plaster cast (a control group) or with a prefabricated functional brace (the Aberdeen Colles' fracture brace). Similar results were obtained in both groups with regard to the reduction and to pain scores but the brace provided better grip strength in the early stages of treatment. This was statistically significant after five weeks for both manipulated and non-manipulated fractures. At the tenth day the results were statistically significant only in manipulated fractures. There was no significant difference in the functional outcome between the two treatment groups. However, younger patients and those with less initial displacement had better functional results. PMID- 12585583 TI - Leakage of cement in percutaneous transpedicular vertebroplasty for painful osteoporotic compression fractures. AB - e analysed the CT scans and radiographs of 76 vertebrae in 49 patients who underwent vertebroplasty for painful osteoporotic compression fractures. Leaks of cement were classified into three types: those via the basivertebral vein (type B), via the segmental vein (type S), and through a cortical defect (type C). More leaks were identified on CT scans than on radiographs by a factor of 1.5 (74/49). Most type-B (93%) and type-S (86%) leaks were missed or underestimated on a lateral radiograph which is usually the only view used during the injection of cement. Of the leaks into the spinal canal, only 7% (2/28) were correctly identified on radiographs. The areas on lateral radiographs where this type of leak may be observed were divided into four zones, and their diagnostic value in predicting a leak into the spinal canal was evaluated. The results showed that cement in the neural foramina had the highest positive predictive value (86%). PMID- 12585584 TI - Intraoperative pulmonary embolism during spinal instrumentation surgery. A prospective study using transoesophageal echocardiography. AB - Intraoperative pulmonary fat and bone-marrow embolism is a serious complication of bone and joint surgery. We have investigated the occurrence and incidence of intraoperative embolism in patients undergoing elective lumbar spinal surgery with or without instrumentation. Sixty adult patients with lumbar degenerative disease were examined by intraoperative transoesophageal echocardiography while undergoing posterior lumbar surgery. Of these, 40 underwent surgery with instrumentation and 20 without. Moderate to severe (grade 2 or 3 according to the grading scale of Pitto et al) embolic events were seen in 80% of the instrumented patients but in none of the non-instrumented patients (p < 0.001). The insertion of pedicle screws was particularly associated with large numbers of pulmonary emboli, while the surgical approach, laminectomy, disc removal and bone harvesting were associated with small numbers of emboli. We consider that, as in arthroplasty and intramedullary fixation of fractures, these embolic events are relevant to the development of potentially fatal fat embolism during spinal surgery. PMID- 12585585 TI - The knee-spine syndrome. Association between lumbar lordosis and extension of the knee. AB - Degenerative changes of the knee often cause loss of extension. This may affect aspects of posture such as lumbar lordosis. A total of 366 patients underwent radiological examination of the lumbar spine in a standing position. The knee and body angles were measured by physical examination using a goniometer. Limitation of extension of the knee was significantly greater in patients whose lumbar lordosis was 30 degrees or less. Lumbar lordosis was significantly reduced in patients whose limitation of extension of the knee was more than 5 degrees. It decreased over the age of 70 years, and the limitation of extension of the knee increased over the age of 60 years. Our study indicates that symptoms from the lumbar spine may be caused by degenerative changes in the knee. This may be called the 'knee-spine syndrome'. PMID- 12585586 TI - Role of posterior stabilisation in the management of tuberculosis of the dorsal and lumbar spine. AB - We present a prospective study of patients with tuberculosis of the dorsal, dorsolumbar and lumbar spine after combined anterior (radical debridement and anterior fusion) and posterior (instrumentation and fusion) surgery. The object was to study the progress of interbody union, the extent of correction of the kyphosis and its maintenance with early mobilisation, and the incidence of graft and implant-related problems. The American Spinal Injury Association (ASIA) score was used to assess the neurological status. The mean preoperative vertebral loss was highest (0.96) in the dorsal spine. The maximum correction of the kyphosis in the dorsolumbar spine was 17.8 degrees. Loss of correction was maximal in the lumbosacral spine at 13.7 degrees. All patients had firm anterior fusion at a mean of five months. The incidence of infection was 3.9% and of graft-related problems 6.5%. We conclude that adjuvant posterior stabilisation allows early mobilisation and rehabilitation. Graft-related problems were fewer and the progression and maintenance of correction of the kyphosis were better than with anterior surgery alone. There is no additional risk relating to the use of an implant either posteriorly or anteriorly even when large quantities of pus are present. PMID- 12585587 TI - Local and systemic control in Ewing's sarcoma of the femur treated with chemotherapy, and locally by radiotherapy and/or surgery. AB - The role of radiotherapy and/or surgery in the local treatment of Ewing's sarcoma has still to be determined. The outcome of Ewing's sarcoma may differ according to its location and a selection bias towards surgery limits the ability to compare methods of local treatment. We have carried out a retrospective review of 91 consecutive patients treated for non-metastatic Ewing's sarcoma of the femur. They received chemotherapy according to four different protocols. The primary lesion was treated by surgery alone (54 patients), surgery and radiotherapy (13) and radiotherapy alone (23). One was treated by chemotherapy alone. At a median follow-up of ten years, 48 patients (53%) remain free from disease, 39 (43%) have relapsed, two (2%) have died from chemotherapeutic toxicity and two (2%) have developed a radio-induced second tumour. The probability of survival without local recurrence was significantly (p = 0.01) higher in patients who were treated by surgery with or without radiotherapy (88%) than for patients who received radiotherapy alone (59%). The five- and ten-year overall survival rates were 64% and 57%, respectively. Patients who were treated by surgery, with or without radiotherapy, had a five- and ten-year overall survival of 64%. Patients who received only radiotherapy had a five- and ten-year survival of 57% and 44%, respectively. Our results indicate that in patients with Ewing's sarcoma of the femur, better local control is achieved by surgical treatment (with or without radiotherapy) compared with the use of radiotherapy alone. Further studies are needed to verify the impact of this strategy on overall survival. PMID- 12585588 TI - Ultrasonographic imaging of fractures of the clavicle in newborn infants. AB - Fractures of the clavicle in the neonate are usually diagnosed by clinical examination and confirmed by plain radiography. Exposure of newborn infants to irradiation should be avoided if possible. Following the clinical examination of 2978 neonates, 15 had suspected fractures of the clavicle. All were confirmed by ultrasound. In combination with clinical examination, ultrasound is a satisfactory alternative to radiological assessment for the diagnosis of fractures of the clavicle in newborn children. PMID- 12585589 TI - Ultrasound in the management of the position of the femoral head during treatment in a spica cast after reduction of hip dislocation in developmental dysplasia of the hip. AB - In this prospective study of 18 hips we compared the efficacy of ultrasound with CT in determining the position of the femoral head in a spica cast after closed or open reduction in children with developmental dysplasia of the hip. Ultrasound was performed through the perineal opening of the cast. With a transinguinal approach, the superior ramus of the pubis, the acetabulum, the femoral head and the femoral neck can be depicted in one plane. The CT and ultrasound images were blinded and reviewed by two of the authors. Ultrasound was inconclusive in the first two reductions since the perineal opening was too small to see all the landmarks in one plane. In the following 16 reductions the landmarks were well defined and interpretation of the CT and ultrasound was similar. The perineal opening in the spica cast should be made in such a way that the ultrasound probe can be positioned in the groin so that the landmarks can be shown in one plane. PMID- 12585590 TI - Delayed ossification of the proximal capital femoral epiphysis in Legg-Calve Perthes' disease. AB - We studied radiographs of 125 children (105 boys, 20 girls) with unilateral Legg Calve-Perthes' disease to examine the epiphyseal development of the femoral head in the contralateral (unaffected) hip. The epiphyseal height (EH) and width (EW) of the unaffected hip were measured on the initial anteroposterior pelvic radiograph. In 109 of the patients (87.2%) the EH was below the mean for normal Japanese children and a significantly small EH (below -2 SDs) was observed in 23 patients (18.4%). By contrast, the EW of most patients (95.2%) lay within +/- 2 SDs of normal values except for six with a significantly small EW. A strong positive linear correlation (R = 0.87) was observed in the EH:EW ratio in the patients. A smaller EH than expected for EW in our series indicated epiphyseal flattening of the femoral head in Legg-Calve-Perthes' disease. Our findings support the hypothesis that a delay in endochondral ossification in the proximal capital femoral epiphysis may be associated with the onset of Perthes' disease. PMID- 12585591 TI - Small volume hypertonic hydroxyethyl starch reduces acute microvascular dysfunction after closed soft-tissue trauma. AB - A major pathway of closed soft-tissue injury is failure of microvascular perfusion combined with a persistently enhanced inflammatory response. We therefore tested the hypothesis that hypertonic hydroxyethyl starch (HS/HES) effectively restores microcirculation and reduces leukocyte adherence after closed soft-tissue injury. We induced closed soft-tissue injury in the hindlimbs of 14 male isoflurane-anaesthetised rats. Seven traumatised animals received 7.5% sodium chloride-6% HS/HES and seven isovolaemic 0.9% saline (NS). Six non-injured animals did not receive any additional fluid and acted as a control group. The microcirculation of the extensor digitorum longus muscle (EDL) was quantitatively analysed two hours after trauma using intravital microscopy and laser Doppler flowmetry, i.e. erythrocyte flux. Oedema was assessed by the wet-to-dry-weight ratio of the EDL. In NS-treated animals closed soft-tissue injury resulted in massive reduction of functional capillary density (FCD) and a marked increase in microvascular permeability and leukocyte-endothelial cell interaction as compared with the control group. By contrast, HS/HES was effective in restoring the FCD to 94% of values found in the control group. In addition, leukocyte rolling decreased almost to control levels and leukocyte adherence was found to be reduced by approximately 50%. Erythrocyte flux in NS-treated animals decreased to 90 +/- 8% (mean SEM), whereas values in the HS/HES group significantly increased to 137 +/- 3% compared with the baseline flux. Oedema in the HS/HES group (1.06 +/- 0.02) was significantly decreased compared with the NS-group (1.12 +/- 0.01). HS/HES effectively restores nutritive perfusion, decreases leukocyte adherence, improves endothelial integrity and attenuates oedema, thereby restricting tissue damage evolving secondary to closed soft-tissue injury. It appears to be an effective intervention, supporting nutritional blood flow by reducing trauma induced microvascular dysfunction. PMID- 12585592 TI - Microvascular response of striated muscle to metal debris. A comparative in vivo study with titanium and stainless steel. AB - Wear products of metal implants are known to induce biological events which may have profound consequences for the microcirculation of skeletal muscle. Using the skinfold chamber model and intravital microscopy we assessed microcirculatory parameters in skeletal muscle after confrontation with titanium and stainless steel wear debris, comparing the results with those of bulk materials. Implantation of stainless-steel bulk and debris led to a distinct activation of leukocytes combined with a disruption of the microvascular endothelial integrity and massive leukocyte extravasation. While animals with bulk stainless steel showed a tendency to recuperation, stainless-steel wear debris induced such severe inflammation and massive oedema that the microcirculation broke down within 24 hours after implantation. Titanium bulk caused only a transient increase in leukocyte-endothelial cell interaction within the first 120 minutes and no significant change in macromolecular leakage, leukocyte extravasation or venular diameter. Titanium wear debris produced a markedly lower inflammatory reaction than stainless-steel bulk, indicating that a general benefit of bulk versus debris could not be claimed. Depending on its constituents, wear debris is capable of eliciting acute inflammation which may result in endothelial damage and subsequent failure of microperfusion. Our results indicate that not only the bulk properties of orthopaedic implants but also the microcirculatory implications of inevitable wear debris play a pivotal role in determining the biocompatibility of an implant. PMID- 12585593 TI - Cell-mediated bioresorption of sintered carbonate apatite in rabbits. AB - Bone apatite contains carbonate and is therefore not pure hydroxyapatite. We have successfully developed sintered carbonate apatite (CA) with a concentration of carbonate of 6 weight% and have evaluated its osteoconductive and bioresorption characteristics. Cylindrical porous sintered CA and sintered hydroxyapatite (HA) measuring 4 x 4 mm with a porosity of 20% were implanted into surgically-created bone defects in the knees of rabbits. The animals were killed after 1, 3, 6 and 12 months. The defects were evaluated by microfocus CT and histology. Bone growth into and around both materials increased. Newly-formed bone was placed in direct contact with both. Osteoclast-like cells resorbed only CA, and were coupled with osteoblasts. The porosity of sintered CA increased, indicating bioresorption, whereas that of sintered HA did not increase. Our findings indicate that sintered CA may be useful as a bioresorbable bone substitute. PMID- 12585594 TI - The Gotfried percutaneous compression plate compared with the conventional classic hip screw for the fixation of intertrochanteric fractures of the hip. PMID- 12585595 TI - Cardiac output during hemiarthroplasty of the hip. PMID- 12585596 TI - Tuberculosis of the thoracic spine. PMID- 12585597 TI - Salivary and serum immunoglobulin levels in cats with chronic gingivostomatitis. AB - The salivary and serum concentrations of immunoglobulins G, M and A (IgG, IgM and IgA), and the salivary concentrations of albumin were measured by ELISA in 30 cats with chronic gingivostomatitis and 32 healthy cats. The cats with chronic gingivostomatitis had significantly higher salivary concentrations of IgG, IgM and albumin, and higher serum concentrations of IgG, IgM and IgA, but significantly lower salivary concentrations of IgA than the healthy cats. The cats with chronic gingivostomatitis were treated with either methylprednisolone, sodium aurothiomalate, metronidazole and spiramycin, or oral hygiene products. After three months of treatment, the cats receiving methylprednisolone had a significant reduction in serum IgG levels compared to the cats treated with sodium aurothiomalate or metronidazole and spiramycin, but after six months of treatment there were no significant differences between the groups. Before the treatments, the levels of oral inflammation were not correlated significantly with any of the serum or salivary immunoglobulin levels. However, the changes in oral inflammation were correlated significantly with the changes in the salivary IgM concentration after three and six months of treatment, and with the change in the salivary IgA concentration after six months of treatment. PMID- 12585598 TI - Efficacy of ivermectin and moxidectin against Otostrongylus circumlitus and Parafilaroides gymnurus in harbour seals (Phoca vitulina). AB - Verminous bronchopneumonia caused by infection with Otostrongylus circumlitus and Parafilaroides gymnurus is an important cause of death during the rehabilitation of harbour seals (Phoca vitulina). During the winter of 2000/01, 35 juvenile harbour seals with severe clinical signs of verminous bronchopneumonia were treated with either 0.2 mg/kg ivermectin orally or 0.2 mg/kg moxidectin subcutaneously, and monitored for 30 days. The efficacy of the anthelmintics was determined by the pattern of larval excretion (Baermannisation) and the progress of the clinical signs. Both anthelmintics had reduced larval excretion by at least 99 per cent after 10 days, but the seals' rapid breathing rate and and dyspnoea returned to normal more quickly in the animals treated with moxidectin. The pharmacokinetics of the anthelmintics were determined by solid-phase extraction, and high-performance liquid chromatography with fluorescence detection. Moxidectin had a mean (sd) residence time of 9.04 (2.12) days compared with 4.83 (1.14) days for ivermectin. PMID- 12585599 TI - Outbreak of clostridial myocarditis in calves. AB - This paper describes the epidemiological and pathological features of an outbreak of clostridial myocarditis in calves due to Clostridium chauvoei. Four of seven two-month-old Hereford calves died in the course of a week. Their gross postmortem lesions were similar and consisted of irregular dark red areas of myocardial necrosis through the full thickness of the atrial and ventricular myocardium. No lesions were observed in skeletal muscle. The heart muscle had extensive multifocal areas of acute coagulative necrosis. The diagnosis was confirmed by a fluorescent antibody technique on tissue smears, by a streptavidin biotin technique on formalin-fixed, paraffin-embedded tissues, and by a PCR technique specific for the 16S rRNA of C. chauvoei. PMID- 12585600 TI - Early prediction of fetal numbers in sheep based on peripheral plasma progesterone concentrations and season of the year. PMID- 12585601 TI - Bacteriological and mycological findings, and in vitro antibiotic sensitivity of pathogenic staphylococci in equine skin infections. PMID- 12585602 TI - Listeriosis in a free-ranging colobus monkey (Colobus guereza caudatus) in Kenya. PMID- 12585603 TI - Chronological antigenic survey of canine parvovirus in Japan. PMID- 12585604 TI - Stingray injury to a horse in coastal waters off eastern Australia. PMID- 12585605 TI - Poor reproductive performance in pigs. PMID- 12585606 TI - Fees for retired RCVS members. PMID- 12585607 TI - Caesarean sections in cows. PMID- 12585608 TI - Imprint cytological examination in PMWS diagnosis. PMID- 12585609 TI - Research into canine diabetes mellitus. PMID- 12585610 TI - Health insurance rebates in sports medicine should consider scientific evidence. PMID- 12585611 TI - The effect of either a pre or post exercise stretch on straight leg raise range of motion (SLR-ROM) in females. AB - This study examined the order effect of a hamstring muscle stretch and resisted hamstring exercises on straight leg raise range of movement (SLR-ROM), in a group of twenty females aged 20-34 years. The stretch was applied either immediately before or Immediately after a resisted hamstring exercise. Exercise significantly decreased SLR-ROM (p < 0.05) and stretching significantly increased SLR-ROM (p < 0.05). The overall change in SLR-ROM for the two groups showed that a significant increase in SLR-ROM was detected (at a low force level) in Group 1 subjects who performed exercise and then stretched. A non-significant increase in SLR-ROM was detected by the higher force level in Group 1 subjects. In contrast, Group 2 subjects (stretch then exercise) showed a non-significant reduction in SLR-ROM at both levels of force. PMID- 12585612 TI - An Australian survey of in-patient protocols for quadriceps exercises following anterior cruciate ligament reconstruction. AB - Current practices regarding in-patient strengthening exercise prescription following anterior cruclate ligament (ACL) reconstruction are scarcely addressed in the literature and remain unreported for Australian hospitals. An observational, questionnaire-based study was used to Investigate in-patient quadriceps strengthening practices in Australian hospitals after ACL reconstruction. Questionnaires were returned by 248 hospitals (76% response rate) and of these 88 hospitals reported performing ACL reconstructions. These hospitals were surveyed to determine the types of quadriceps strengthening exercises prescribed during the in-patient period. Information was sought regarding routine management strengthening practices. A variety of quadriceps exercises such as static quadriceps contractions (SQC), straight leg raises (SLR) and inner range quadriceps (IRQ) were frequently prescribed, although no standard practice currently exists. Static quadriceps exercises were prescribed by 91% of hospitals surveyed, IRQ exercises were used by 30%, and 49% reported using SLR exercises. Current literature tends to support the performance of SQC and SLR exercises during the early postoperative period following ACL reconstruction. Conversely, current clinical practice is incongruent with regard to the scientific recommendations for IRQ performance. PMID- 12585613 TI - The effects of therapeutic massage on delayed onset muscle soreness and muscle function following downhill walking. AB - This study Investigated the effects of a therapeutic massage on delayed onset muscle soreness and muscle function following downhill walking. Eight male subjects performed a 40-min downhill treadmill walk loaded with 10% of their body mass. A qualified masseur performed a 30-min therapeutic massage to one limb 2 hours post-walk. Muscle soreness, tenderness, isometric strength, isokinetic strength, and single leg vertical jump height were measured on two occasions before, and 1, 24, 72 and 120 hours post-walk for both limbs. Subjects showed significant (p < 0.004) increases in soreness and tenderness for the non-massaged limb 24 hours post-walk with a significant (p < 0.001) difference between the two limbs. A significant reduction In isometric strength was recorded for both limbs compared to baseline 1 hour post-walk. Isokinetic strength at 60 degrees/sec and vertical jump height were significantly lower for the massaged limb at 1 and 24 hours post-walk. No significant differences were evident in the remaining testing variables. These results suggest that therapeutic massage may attenuate soreness and tenderness associated with delayed onset muscle soreness. However it may not be beneficial in the treatment of strength and functional declines. PMID- 12585614 TI - The epidemiology of medically attended sport and recreational injuries in Queensland. AB - The Queensland Sport and Recreation Injury Survey (QSRIS) is a retrospective study describing the annual incidence of injuries in the state of Queensland, Australia, resulting from sport and recreational activity involvement. Data were collected by means of a computer-assisted-telephone-interview (CATI) survey of a representative sample of Queenslanders in the spring of 2000. The sample produced a total of 1337 respondents aged 18 to 94 years. The survey asked information regarding medically attended, non-fatal injuries resulting from sport and recreational activities in the past 12 months. Of the 1337 individuals surveyed, 191 of the respondents reported one or more injuries that required medical attention resulting in a total of 222 Injuries. This represents an overall rate of 1,666 medically attended injuries per 10,000 people. Among those reporting a sport or recreational injury, the most common types of injuries were a strained/pulled muscle (30.9%), sprained/torn ligament (24.1%), and fracture (12.6%). The most common bodily locations of injuries were the shoulders (13.0%) and knees (12.5%). Results detail the nature and type of injury, medical professional attending to the injury and the nature of the sport or recreational activity that led to the reported injury, highlighting the number of injuries associated with general fitness activities achieved through high participation rates and low injury rates. This remains an area of much needed attention given the promotional push towards raising the levels of physical activity at a population level. PMID- 12585615 TI - A comparison of two injury surveillance systems within sports medicine clinics. AB - This study compares a comprehensive method of collecting injury data from sports medicine clinics, with a more simplified method of injury surveillance. The sports medicine injury surveillance (SMIS) project was implemented in a group of five allied sports medicine clinics in Melbourne. over two consecutive years. The injury surveillance method used in the second year (SMIS2) was a simplified version of that used in the first year (SMIS1). Methodological differences in the injury surveillance systems included form design, staff commitment and training, auditing process, financial incentives offered and employment of a project officer. Data were collected on 6479 new sports injury patients during SMIS1 and on 1682 patients during SMIS2. Comparative data from the two years of injury surveillance included patient profile (gender. age. days from injury to treatment, sport and context of injury) and injury information (site, cause and nature of injury). The SMIS2 methodology was associated with a lower sensitivity (p < 0.001) and a higher proportion of missing information (p < 0.001) than the SMIS1 methodology. There was also a significant difference in the nature and cause of injury variables (p < 0.001) between SMIS1 and SMIS2 and this was associated with coding changes. This study shows that the method of data collection influences both the proportion of missing information and the sensitivity of the system. A comprehensive method of injury surveillance will lead to a more complete data collection process. Methodological differences, however, do not appear to substantially alter conclusions about general patient characteristics, but do have some influence on the accuracy with which broad injury data are identified. Notwithstanding these comments, this study shows that injury surveillance activities can be successfully implemented in sports medicine clinics. PMID- 12585616 TI - Five errors about error. PMID- 12585617 TI - High intensity exercise assessment: relationships between laboratory and field measures of performance. AB - The purpose of this study was to examine the relationship between high intensity cycle ergometry when resistive forces were optimised and field tests of high intensity performance in an elite sprint trained group (n = 12). Body mass, stature and age of the group were determined prior to testing (66.3 +/- 9.8kg, 177 +/- 7.6cm and 17 +/- 0.74 yrs respectively). Field tests of high intensity performance examined included sprinting. maximal shuttle running and jumping ability (vertical and horizontal). Significant correlations existed between all field tests (P < 0.01). Only moderate correlations were observed for values generated on the cycle ergometer and performance tests (P > 0.05). The results of the present study suggest that the optimised cycle ergometer test and the performance tests are unrelated when evaluated statistically. These findings also suggest that the sprinting and jumping tests examined are highly related and may be substituted for each other as possible predictors of high intensity ability. PMID- 12585618 TI - A valid and reliable method for measuring the kicking accuracy of soccer players. AB - Kicking accuracy is an important component of soccer performance. We constructed a plywood target measuring 243.5cm wide x 122cm high. Carbon paper applied to the surface of the target allowed measurement by 2 raters from a bull's-eye to 10 ball marks left by kicks. Intraclass correlation coefficients with 95% confidence intervals were used to determine the intra- and interrater reliability of the measurement to each ball mark. Mean and median distances from bulls-eye to ball mark were 89.9cm and 97.9cm, respectively (range, 25.7 to 150.75cm). The intraclass correlation coefficients for intra- and interrater reliability were 0.99. The root mean square error of measurement indicated that accuracy of measurement was within 0.15cm. These results suggest that our method of assessing kicking accuracy is a valid and reliable tool for analysing performance. Because this tool closely replicates kicking into a soccer goal, we feel that it also has validity. To our knowledge, no other tool or measure (e.g.. number of shots on goal or number of goals per game) has comparable validity and reliability. This method can be used as a training tool and for future investigations of kicking accuracy. PMID- 12585619 TI - Reliability of performance in repeated sprint cycling tests. AB - We have estimated the reliability of performance in a commonly employed exercise test consisting of repeated sprints on a cycle ergometer. Eight recreationally active young men completed a practice trial and three more trials at 3- to 6-day intervals. Each trial consisted of two bouts of 30-s maximal-effort cycling on an electromagnetically braked cycle ergometer; the bouts were separated by 4 min of rest. The typical (standard) errors of measurement for peak and mean power between trials 2 to 4 were 2.5 and 1.7% respectively for the first bout and 1.9 and 1.8% for the second bout. These errors are substantially less than those in previous reliability studies of single 30-s sprint tests, probably because of differences in quality of ergometer. The typical errors for the difference between bouts (i.e., fatigue) for peak power and mean power were 3.0 and 2.5%, respectively. Typical errors for the average of the two bouts were 1.6 and 1.2% for peak and mean power respectively, which are small enough to give adequate precision for moderate treatment effects in studies with modest sample sizes. PMID- 12585620 TI - Validity and reliability of a voice-recognition game analysis system for field sports. AB - The purpose of this study was to assess the ability of observers to use voice recognition analysis to accurately classify gait transitions and quantify gait durations typical of team games. Inter-rater and intra-rater reliability was also determined. Four males were filmed performing pre-determined gait protocols. each comprising different sequences of walking, jogging, running and sprinting. Two operators independently classified gait transitions and the time spent in each gait was determined by the voice recognition system. All gait modes as measured by trained observers demonstrated statistically significant correlations (p < 0.01) to pre-determined measurement criteria. The mean absolute error for all gait transitions was less than half a second (0.32-0.36 s) with the maximum percentage error being approximately 4% for the walk, jog and run gaits and 10% for sprinting. Gait classification error was low at 1.9%. The intra-rater and inter-rater reliability was consistently high ranging from r = 0.87 to 0.99. In conclusion, observers using voicerecognition software provided valid measures of time spent in each of the four gait categories with 90% or better accuracy achieved. PMID- 12585621 TI - Re-assessing accumulated oxygen deficit in middle-distance runners. AB - The purpose of this study was to re-assess the accumulated oxygen deficit (AOD), incorporating recent methodological improvements i.e., 4 min submaximal tests spread above and below the lactate threshold (LT). We Investigated the Influence of the VO2 -speed regression, on the precision of the estimated total energy demand and AOD. utilising different numbers of regression points and including measurement errors. Seven trained middle-distance runners (mean +/- SD age: 25.3 +/- 5.4y, mass: 73.7 +/- 4.3kg. VO2max 64.4 +/- 6.1 mL x kg(-1) x min(-1)) completed a VO2max, LT, 10 x 4 min exercise tests (above and below LT) and high intensity exhaustive tests. The VO2 -speed regression was developed using 10 submaximal points and a forced y-intercept value. The average precision (measured as the width of 95% confidence Interval) for the estimated total energy demand using this regression was 7.8mL O2 Eq x kg(-1) x min(-1). There was a two-fold decrease in precision of estimated total energy demand with the Inclusion of measurement errors from the metabolic system. The mean AOD value was 43.3 mL O2 Eq x kg(-1) (upper and lower 95% CI 32.1 and 54.5mL o2 Eq x kg(-1) respectively). Converting the 95% CI for estimated total energy demand to AOD or including maximum possible measurement errors amplified the error associated with the estimated total energy demand. No significant difference in AOD variables were found, using 10,4 or 2 regression points with a forced y-intercept. For practical purposes we recommend the use of 4 submaximal values with a y-intercept. Using 95% CIs and calculating error highlighted possible error in estimating AOD. Without accurate data collection, increased variability could decrease the accuracy of the AOD as shown by a 95% CI of the AOD. PMID- 12585622 TI - The impact of interventional neuroradiology on neurosurgical training. AB - Aneurysmal subarachnoid haemorrhage carries a high mortality and morbidity. Surgical treatment (craniotomy and clipping of the aneurysm) has been, until recently, the gold standard treatment. Endovascular embolisation treatment has rapidly evolved and the evidence available suggests that the results are as good as surgery. Endovascular treatment successfully occludes the aneurysm to prevent re-haemorrhage, whilst reducing the procedural morbidity when compared to craniotomy and clipping. It is perceived to be of particular benefit for aneurysms in the posterior cerebral circulation where operative morbidity and mortality are significantly higher than for aneurysms on the anterior circle of Willis. The establishment of endovascular treatment has reduced the number of cases being treated surgically, and this has had a significant effect on surgical training. We analysed the management of all ruptured aneurysms treated in our unit over a 4-year period. During the same period, an endovascular service was established in the unit. We devised a novel system for the angiographic grading of aneurysms in order to evaluate the impact that coiling has had on surgical training. The results show that as few as four aneurysms per year would be appropriate for specialist registrars to operate upon. We propose some mechanisms for maintaining high quality surgical training. PMID- 12585623 TI - Laparoscopic fundoplication: learning curve and patient satisfaction. AB - AIMS: Laparoscopic fundoplication is now accepted as the optimal surgical option for the management of selected cases of gastro-oesophageal reflux disease. The principal aim of this study was to evaluate the learning curve experience of two consultant surgeons in the technique of laparoscopic fundoplication (LF). Additional variables assessed were total number of cases, preoperative investigations, conversion rate, duration of operation, ASA grade, morbidity, mortality, necessity of further procedures, and patient satisfaction rate. PATIENTS AND METHODS: Retrospective case-note analysis of all adult patients who underwent fundoplication under the care of two consultant general surgeons over a 3-year period from January 1997 to December 1999. RESULTS: A total of 61 patients were included, 31 males and 30 females, with a median age of 46 years (range, 21 73 years). Of the patients, 90% were either ASA 1 or 2. The mean time for which the 24-h pH < 4 was 20.5% (95% CI, 15.3-25.7). Of the 61 patients, 6 were operated on by open technique, for medical reasons and previous abdominal procedures. Out of the remaining 55 patients, 13 had to be converted (23.6%). Mean operating times were 120 min for LF, 85 min for open operation and 142 min for LF plus conversion. There was a significant decline in conversion rate with time (P < 0.002). Mortality was nil. One patient had a perforation of the cricopharyngeus secondary to insertion of a bougie. The mean length of hospital stay following the laparoscopic technique was 3.4 days compared to 8.7 days following the open technique. Overall, 59 patients (96%) were happy with the result, and the operation failed in 2 patients. Five patients (8%) needed endoscopic dilatation in the first few weeks after the operation. CONCLUSIONS: The results show that LF is a safe procedure, takes longer than open procedure, and has an acceptable morbidity. Experience with the technique reduces the need for conversion. The mean length of hospital stay is significantly less and there is a high level of patient satisfaction. PMID- 12585624 TI - Surgical OGD--a dying art? AB - INTRODUCTION: Reductions in surgical training and the increases in medical gastroenterology have raised concerns that surgeons may not be adequately trained in upper gastrointestinal (GI) endoscopy. METHOD: To evaluate this problem, a questionnaire was sent to all current surgical specialist registrars (SpRs) in the South Thames East Region. RESULTS: There was an 82% (52/63) response rate. Only 50% (26/52) of trainees had received more than 6 months' training in upper GI endoscopy. 83% (43/52) were in posts which did not provide adequate elective exposure for training. 50% (26/52) were required to provide an emergency service, despite a paucity of experience and less than 50% were able to perform therapeutic injection. In the main, emergency endoscopy is performed with substandard equipment, poor facilities, and untrained staff. CONCLUSIONS: Surgical trainees are poorly trained and do not have the necessary skills to provide an emergency service for upper GI haemorrhage. Emergency endoscopy facilities are severely under resourced. PMID- 12585625 TI - A prospective study of 1000 hernias: results of the Plymouth Hernia Service. AB - BACKGROUND: A hernia service within a general hospital was prospectively evaluated to establish whether evidence-based protocols could deliver results comparable to those reported from specialist hernia clinics. METHODS: Protocols were devised according to established models. With the support of a nurse specialist, 1015 patients with inguinal hernia were treated. Quality-of-life analysis was undertaken using the Short Form 36. RESULTS: Patients ranged in age from 16-98 years (median, 56 years). Ambulatory day-case surgery was achieved in 820 patients (81%), with local anaesthesia in 891 (88%). Wound infection occurred in 10 patients (0.98%). Wound haematoma requiring surgical intervention occurred in three patients. Two patients formed wound seromas that settled spontaneously. One patient developed ischaemic orchitis resulting in testicular atrophy. At 5 days after operation, 91% of patients had returned to normal activity. At 1 year, 7 patients (0.7%) had pain sufficient to limit normal activity or employment. There were 8 recurrences (0.78%) at a median follow-up of 2.5 years. Quality-of life was enhanced at 1 year postoperatively. CONCLUSION: A protocol-driven hernia service within a general hospital can provide patient outcomes comparable to specialist hernia clinics. PMID- 12585626 TI - Reliability of pre-operative biopsies in the histological grading of colorectal adenocarcinomas. AB - BACKGROUND: Management of colorectal cancer depends on many patient and tumour variables. Decisions are sometimes based on the histological grade of tumour as assessed by initial biopsy. This retrospective study looked at the reliability of grading colorectal cancer from pre-operative biopsies. METHODS: 118 patients with a diagnosis of colorectal cancer, who had adequate pre-operative biopsies, were included in the study. A single consultant histopathologist re-examined all the preoperative biopsy slides, but was blinded to the final grading of the resection specimen. RESULTS: Of the 118 resection specimens, 41 were graded well differentiated, 52 moderately differentiated and 25 poorly differentiated. When compared with postoperative grading, 37 (31%) initial biopsies were undergraded, 49 (43%) were correctly graded and 32 (26%) were overgraded. CONCLUSIONS: These results suggest that the histological grading of pre-operative specimens is inaccurate and should be used with caution to determine treatment. The difficulty with grading pre-operative biopsies in colorectal cancer may be owing to a lack of specific criteria such as is used in, for example, breast cancer. PMID- 12585627 TI - Should 'doughnut' histology be routinely performed following anterior resection for rectal cancer? AB - Stapled low anterior resection is widely employed in the treatment of rectal adenocarcinoma. The technique yields two tissue 'doughnuts' which are often submitted for histological examination. This process is labour intensive and not part of the minimum data set for colorectal cancer histopathology reports. A consecutive series of anterior resection doughnuts from 125 patients was reviewed retrospectively to assess the impact of doughnut pathology on the management of patients. Four doughnuts had a histological abnormality reported but none of these altered treatment. Routine histological examination of 'doughnuts' is not beneficial to the management of patients undergoing surgery for rectal adenocarcinoma. PMID- 12585628 TI - The development of a protocol in using day surgery for minor orthopaedic trauma patients. AB - We examined all minor orthopaedic trauma patients requiring surgical intervention at Manchester Royal Infirmary over a 6-month period to determine whether minor trauma patients meet the day surgery requirements in terms of medical fitness, analgesia requirements and postoperative complications. A total of 83 patients had surgery for minor orthopaedic trauma in the defined period. Of these patients, 79 (95.2%) were medically fit, 45 (70%) were admitted to a hospital ward and 19 (30%) were brought back to day surgery. None of the patients attending day surgery developed postoperative complications or required return to hospital. The majority of patients admitted to a ward required simple or no analgesia pre- (95.5%) and postoperatively (100%), and most were discharged on the same (55.6%) or next (35.6%) day of their operation. We suggest a protocol whereby patients with minor trauma are brought back to day surgery. This could potentially reduce pressures on bed availability. PMID- 12585629 TI - An audit of the out-patient follow-up of hip and knee replacements. AB - Medical records of 100 consecutive patients who underwent hip and knee replacements (56 hips and 44 knees) in 1997-1998 were studied. Particular attention was paid to the out-patient follow-up appointments with regard to any postoperative complication and intervention based on the clinical and radiological assessment conducted during the follow-up visit. The average period of follow-up was 845 days. During this period, these patients had 304 out-patient visits. Twenty-two patients had a problem during this period of whom 10 needed an intervention. Of the 10 patients who needed an intervention, 3 were identified during the routine out-patient visits whereas 7 were either by general practitioner referral or were seen in the accident and emergency department. At these appointments, 187 radiographs were taken. Of these, 8 (4.3%) showed some abnormality. The minimum cost for these follow-up appointments was estimated to be 23,297 pounds sterling. We recommend that the postoperative out-patient appointment of the patients with hip and knee replacements should be restricted to a visit at 6-12 weeks followed by discharge if no problems are anticipated. PMID- 12585630 TI - Research output of a cohort of orthopaedic consultants in Great Britain. AB - There has been renewed emphasis on research following the Calman recommendations for higher training. It is often considered essential for progression in training to have participated in research projects and to have some publications to one's credit. Orthopaedic surgery is usually thought to lag behind other disciplines in this respect. It was decided to conduct a review of the research output of a cohort of orthopaedic consultants in Great Britain to obtain baseline information for future comparisons. A group of surgeons who graduated at least 25 years ago was chosen as it was felt this would provide the pattern of research output throughout the surgical career of the cohort. The type of publication and the various factors that may affect research output were studied. There was a significant difference in the output of consultants working in teaching hospitals and large centres compared to those in district general hospitals. Is the soil as important as the seed? PMID- 12585631 TI - Surgical time and motion: the intermediate equivalent revisited. AB - The relationship between operative time, the intermediate equivalent value (IEV) and the complexity of common general surgical operations was examined. Correlation was found between the BUPA schedule values for procedures categorized as intermediate and major, but complex major vascular reconstruction and oesophagogastric resection for cancer occupied significantly more theatre time than the four intermediate equivalents allocated by the Collins or BUPA schedule. Moreover, anaesthetic preparation time for complex major surgery in the latter surgical subspecialities contributed at least one further intermediate value. Re evaluation of the ideal IEV weighting of all surgical operations including anaesthetic input from larger similar audits would allow more accurate audits of surgeons' work-load, and also facilitate transparent intensive management of operating theatre resource. PMID- 12585632 TI - Finding the best from the rest: evaluation of the quality of patient information on the Internet. AB - INTRODUCTION: Information is of utmost importance for patients at risk of developing cancer who require regular screening. Quality assessment is vital to ensure correct information is published on the Internet. METHOD: A postal questionnaire was sent to patients under follow-up for Barrett's oesophagus and colonic polyps. Questions related to computer/Internet access, where patients had previously sought information, whether web-sites would be of use, and what information they would like displayed. A review of on-line patient literature for Barrett's oesophagus and colorectal adenomas was performed. RESULTS: Of the 200 questionnaires sent, 161 patients responded (80.1%). The majority of patients (88%, n = 141) wanted more information on their condition, with 45% (73) having home Internet access and a further 32% (52) having web access from other sources. Only 8% (12) had used the Internet as a source of information; however, the majority of patients (57%) would access a recommended web-site. The Barrett's search resulted in 10/200 sites with full information (i.e. score > 8/10 points). For colorectal polyps there were 12/200 sites. CONCLUSIONS: Accessing quality Internet health information is very time consuming. Recommended web-sites that provide the best information would help patients avoid being overwhelmed with irrelevant and confusing literature. PMID- 12585633 TI - Microdochectomy in women aged over 50 years. AB - Management of a spontaneous single duct nipple discharge without associated mass and normal mammography remains controversial. Our study examined the pathological results of 52 women of all ages treated with microdochectomy for single duct nipple discharge. Malignant or premalignant lesions were identified in 1 patient under 50 years of age and 2 patients over 50 years. Papilloma was the most frequently identified pathology in both age groups (60% of patients under 50 years and 73% over or equal to 50 years of age). Our results suggest that microdochectomy is a safe effective treatment in women aged over 50 years. PMID- 12585634 TI - Retrograde embolisation of varicocele in the paediatric age group: a review of 10 years' practice. AB - OBJECTIVE: To review the 10 years' practice of retrograde embolisation of varicoceles in the paediatric age group. PATIENTS AND METHODS: 40 boys (age range, 6-16 years), with varicocele diagnosed both clinically and sonographically, were referred to the radiology department for embolisation. Data of these patients were collected and analysed retrospectively. Patients' age, side, treatment, complications, and outcome were noted. The follow-up ranges from 2 months to 6 years (mean, 1.1 years). RESULTS: In 24 patients (60%), embolisation was successful at the first attempt and 11 procedures (27.5%) were technical failures; 4 patients (10%) after venography were considered non embolisable. One patient (2.5%) is yet to be seen in the clinic. Recurrence occurred in 4 patients (10%) of whom 3 had successful repeat embolisation. One patient with a recurrent varicocele did not wish further treatment. Out of the 11 failures, 5 underwent repeat embolisation with success in 3 patients and failure in two. Four patients had successful surgical ligation and 2 patients were treated conservatively. All who were non-embolisable were treated successfully by surgery. Our overall success rate was 65% (n = 26). CONCLUSIONS: These results suggest that embolisation is valuable as a first line of treatment for paediatric varicoceles provided a skilled interventional radiologist is available. This procedure is less invasive, avoids general anaesthetic, involves minimal postoperative pain, and allows an earlier return to normal activities. However, 35% of patients did eventually require conventional surgery for primary failure of embolisation or late recurrence. PMID- 12585635 TI - The outcome of varicose vein surgery at 10 years: clinical findings, symptoms and patient satisfaction. AB - BACKGROUND: Data on long-term outcomes of varicose vein surgery are sparse, and 'success' rates vary substantially depending on the method of assessment. This study used a variety of methods to evaluate patients 10 years after operation. METHODS: A consecutive cohort of 100 patients was identified, of whom 70 (151 operated limbs) were reviewed--50 by clinical and Doppler examination, and all by structured questionnaire. RESULTS: A few months after operation, symptoms were 'cured' or 'much better' in 89% of patients, and remained so at 10 years in 77%. Only 30% patients were completely free of recurrent varicose veins at 10 years (both by self- and clinician-assessment): 44% had 'just a few' varicosities while 26%, had varicose veins 'as badly as before'. Only 24 patients (34%) were not 'generally pleased' at 10 years, because of continuing symptoms (3), recurrent varicosities (13), and other miscellaneous reasons. CONCLUSIONS: Although some recurrence of varicose veins is frequent 10 years after operation, surgery provides long-term relief of symptoms in the great majority of patients. PMID- 12585636 TI - Assessment of the ischaemic leg. PMID- 12585637 TI - Obturator 'kick'--how to avoid an own-goal! PMID- 12585638 TI - Obturator hernioplasty: a new operative approach and technique of repair. PMID- 12585639 TI - Tegaderm technique to protect skin marking. PMID- 12585640 TI - Orientation and labelling: use of an acetate sheet to label tumour resection specimens. PMID- 12585641 TI - Flexion contracture deformities associated with circular frames--an alternative form of splintage. PMID- 12585642 TI - Goggles are more important than face masks. PMID- 12585643 TI - Blind percutaneous insertion of Hickman lines by a specialist team. PMID- 12585644 TI - Consequences of prolonged wait before gallbladder surgery. PMID- 12585645 TI - A comparison of junior hospital doctors' interpretation of acute radiographs using an X-ray box and a window. PMID- 12585646 TI - Can the tips of marker pens act as a source of cross-infection? PMID- 12585647 TI - Mutations of BRCA genes in hereditary breast and ovarian cancer. AB - BRCA1 and BRCA2 are the main genetic elements associated with hereditary susceptibility to breast and ovarian cancer that have been isolated to date. DNA analysis can identify carriers of mutations in these genes, who, accordingly, can benefit by adequate clinical programs for the reduction of cancer risk. Unfortunately, the analysis is complicated by the complexity of associated mutation spectra. The procedure for mutation testing in BRCA genes and its limitations are discussed. PMID- 12585648 TI - Medical counselling in the clinical management of BRCA mutation. AB - Among medical activities, the choice of an intervention is less and less the primacy of experts over persons, whom under the current dominant model of autonomy are the ones in charge of the decision. However, counselling is ethically relevant and acceptable provided that three conditions are fulfilled: 1. Besides information, patient asks for advice; 2. Advice is not given in an authoritative way, with strong incitement or directiveness; 3. It should be explicitly expressed that, if the patient doesn't follow the physician's advice, this will by no means prevent treatment nor the continuation of their relationship. Within an utilitarian perspective, a decision is something like a balance of facts and values. Therefore, the main (insurmountable?) difficulty lays in the assessment of patient's values. Counselling is all the more difficult and should be cautiously delivered as the situation at stake is assumed to exhibit a huge range of inter-individual values. Physicians should have deep respect for genuine individual values and/or cultural factors. The aim of clinical management is to increase life expectancy, taking into account quality of life and dignity of the person. Deliberative, iterative process seems to be a good way to reach a fair decision. While counselling the patient, the physician has to know for whom (or what) (s)he is acting. Indeed it is not always easy to distinguish among the interest of the patient, of its relatives, of the community of patients or even the interest of science or... self-interest. PMID- 12585649 TI - Hereditary risk of breast cancer: not only BRCA. AB - The BRCA1 and BRCA2 genes are involved in genetic susceptibility to breast cancer (BC). Nevertheless, in a relevant number of families displaying a disease pattern suggesting an inherited susceptibility to BC, mutational analysis fails to detect any defect in the BRCA genes. Therefore, women belonging to such families should be considered eligible for programs aimed at BC control in individuals at hereditary risk. A clinico-mammographic surveillance program for women at high genetic risk, as defined on the basis of pedigree, has been carried out at our centre for ten years, leading to the diagnosis of 19 new BC cases. Only in 13% of the families analysed, the underlying genetic defect was evidenced in BRCA1 or 2. Here we describe two BC prone families where, although no mutations were detected in BRCA genes, follow-up confirmed an increased BC incidence. In three women belonging to these families clinico-mammographic surveillance resulted to be successful in detecting early-stage BC, supporting the usefulness of screening women from high-risk families, irrespective of whether a mutation was found. PMID- 12585650 TI - Evaluating breast cancer risk: available models to assess individual breast cancer risk and probability to be a BRCA mutation carrier. AB - Several models have been generated to estimate breast cancer risk and to predict the probability to identify a BRCA1 or BRCA2 germ line mutation. Each model, based on a slightly different study design and purpose, takes into account a definite number of relevant parameters and ultimately pertains to a specific cohort of women or population. Aim of this work is to underline validity and limits of available tools and their appropriate use in the daily practice. PMID- 12585651 TI - Perspectives in the clinical management of BRCA mutations carriers. AB - The recent identification of the breast cancer-associated genes BRCA1 and BRCA2 is changing the clinical care provided to women at high-risk of breast cancer. We briefly review what is currently known about the clinical management of individuals who bear (or are suspected of bearing) genes mutations and which are the prevention strategies that would reduce the incidence and mortality of breast cancer in this subset of women. PMID- 12585652 TI - Tumour detection and staging by contrast enhanced MRI. PMID- 12585653 TI - Assessing response to treatment in breast cancer using magnetic resonance. AB - Magnetic resonance imaging provides a range of powerful techniques for assessing breast cancer. Contrast enhanced MRI has been shown to provide a sensitive method for assessing primary breast cancer, with increasing emphasis on performing dynamic analysis, and using quantitative analysis techniques. Magnetic resonance spectroscopy provides information on tumour metabolism and has been investigated as a method of assessing response to therapy. The response of metastatic disease from bone cancer has been assessed using T1 weighted methods. MRI provides information on response to treatment early in the course of chemotherapy. Following chemotherapy it provides a sensitive method of assessing residual disease. Both, with conventional treatments and with new antiangiogenic, antivascular and cytostatic treatments, there is a growing role for functional methods of assessing tumour response to treatment. PMID- 12585654 TI - Magnetic resonance imaging of breast cancer angiogenesis: a review. AB - The development and growth of solid tumors rely on a process known as perfusion which allows for the delivery and clearance of nutrients through their vasculature. The classical approach of studying tumor vasculature by histologic staining of endothelial cells provides a measure of microvessel density in areas of high vascularization (so-called "hot spots"). More advanced high-resolution, parametric, contrast-enhanced MRI techniques enable quantitative assessment of the vascular distribution and function over the entire tumor. Hence, MRI overcomes the intratumoral variation of the histological method. Clinical testing of the MRI approach, that evaluates the permeability and surface area of the tumor vasculature, demonstrates improved accuracy of breast cancer diagnosis. PMID- 12585655 TI - Present indications for the use of breast MRI. PMID- 12585656 TI - New technologies in breast MRI. PMID- 12585657 TI - Interventional breast MRI: needle localisation and core biopsies. AB - MRI of the breast is increasingly used to detect additional, conventionally occult primary or recurrent breast cancer. To enable a safe and yet tissue sparing histologic sampling of these lesions, techniques are needed to allow the preoperative wire localization or core biopsy under MR guidance. This text serves to provide an overview on the current status of the different approaches that have been developed to allow selective histologic sampling of "MR-only" lesions. PMID- 12585658 TI - Contrast agents and temporal resolution in breast MR imaging. AB - The article summarizes the technique of dynamic Gd-enhanced breast magnetic resonance (MR) imaging and the established clinical indications to this examination. The main features of the four different non-selective Gd-chelates today available for contrast-enhanced MR breast imaging are described. Their typical two-compartment (vascular/interstitial) distribution is related to the temporal resolution used for the MR acquisition; the gain in specificity obtained increasing the temporal resolution during the first phase after the contrast intravenous injection (using both, rapid T1-weighted sequences or ultrafast T2* weighted perfusion imaging) is discussed. Moreover, a survey of new contrast agents is supplied, including a high T1-relaxivity Gd-chelate, blood pool agents, and lymph node targeted agents. In conclusion, future directions are delineated. PMID- 12585659 TI - Maximum intensity projection analysis in magnetic resonance of the breast. AB - The aim of this study was to determine the sensitivity, specificity and accuracy of vascular asymmetries and vessel location seen using MR Maximum Intensity Projection imaging for malignant and benign lesions. We enrolled 101 women, with an age ranging from 29 to 69 years (mean age of 53.1 years), with a mammary nodule and candidate for surgical intervention. MR was carried out using a 1.5 Tesla superconductive magnet, equipped with a double and bilateral Phased Array dedicated coil. All images obtained were processed using MIP. The relationship between an asymmetry due to a greater amount of vascular structures in the breast with a malignant neoplasm and symmetry with benign neoplasms were statistically significant (p < 0.001): sensitivity of 71.8%, specificity 100% and accuracy 76.2%. The relationship between perilesional or intralesional vessels with malignant versus benign neoplasms were statistically significant (p < 0.001): sensitivity 88.5%, specificity 82.6% and accuracy 87.1%. We concluded that asymmetries between breasts and the presence of perilesional or intralesional vessels seen in MIP images are important signs and, therefore, must be reported. PMID- 12585660 TI - Magnetic resonance of the breast: correlation between enhancement patterns and microvessel density in malignant tumors. AB - The continuous growth of a solid tumor is contingent upon the induction and formation of new vascular structures (angiogenesis). Several studies have established a close correlation between the microvessel density (MVD) of a tumor and its growth capacity. Therefore, a non-invasive method to accurately assess tumor MVD in vivo would be clinically useful. We studied 20 consecutive patients (age range: from 39 to 78 yrs; average age: 58.9 yrs), with malignant breast neoplasms using dynamic MR examinations with a paramagnetic contrast agent and post-operative pathological evaluation. We found that the differences in dynamic MRI enhancement are a direct consequence of microvascular distribution within the neoplasm. This study did not yield a statistically significant correlation between high microcirculation neo-formation and high aggressiveness of the neoplasm, but did indicate that further studies with a larger study populations are necessary. PMID- 12585661 TI - Ductal carcinoma in situ: is there a role for MRI? AB - The purpose of our study is to report personal experience about 28 cases of Ductal Carcinoma In Situ (DCIS) studied with Magnetic Resonance Imaging (MRI). From September 1995 to December 2001, 28 women affected by DCIS lesions underwent contrast enhanced MRI. All patients were submitted previously to mammographic examination. The results of histopathological examination included: 19 DCIS and 9 DCIS with associated microinvasive component or microfoci of invasive ductal carcinoma (IDC). MRI was performed with a 1 T system. A three dimensional fast low-angle shot (FLASH) pulse sequence was used, with a repetition time (TR) of 14 ms, an echo time (TE) of 7 ms and a flip angle of 25 degrees. We evaluated the morphologic features of the enhancement, the enhancement rate and the signal time intensity curve. On MRI 26 out of 28 (92.85%) DCIS lesions showed contrast uptake. 17 out of 19 pure DCIS lesions demonstrated contrast enhancement: 5 showed a low, 7 indeterminate and 5 a strong enhancement. Morphologically, the enhancing lesion was focal in 9, segmental in 5 and linear branching in 3. Wash out was found in 5 cases, plateau curve in 9 and Type I curve in 3. There were 5 cases of multifocality. All DCIS with associated microinvasion demonstrated contrast enhancement: 2 out of 9 cases showed a low enhancement, 4 out of 9 showed an indeterminate enhancement and 3 out of 9 showed a strong enhancement. Morphologically the enhancing lesion was focal in 3 out of 9, segmental in 5 and linear branching in 1. The wash out was demonstrated in 4 cases out of 9, plateau curve in 4 and Type I curve in 1. There were 3 cases of multifocality. In conclusion, MR imaging sensitivity for DCIS detection is lower than that achieved for invasive breast cancer; however, contrast enhanced MR imaging can depict mammographically occult foci of DCIS. The MR imaging technique is of complementary value in better description of tumor size and in additional malignant lesions detection. PMID- 12585662 TI - The evaluation of women with familial risk of breast cancer. AB - The life-time risk for getting breast cancer in The Netherlands is about 11%. About 5% to 10% of these cases are hereditary. Because the value of mammography especially in younger women is limited, the significance of MRI of the breasts in this special group of women is evaluated. PMID- 12585663 TI - High-risk screening: multi-modality surveillance of women at high risk for breast cancer (proven or suspected carriers of a breast cancer susceptibility gene). AB - Familial breast cancer accounts for about 10% of all breast cancer cases. Subjects with mutations in one of the breast cancer susceptibility genes face an about 90% lifetime risk to develop breast cancer. In addition, more than half of the patients will develop breast cancer already before the age of 50, and a significant number even before the age of 35. Screening for familial breast cancer needs to start, therefore, at the age of 30. Detection and differential diagnosis of breast cancer in such a young patient cohort is difficult to achieve with mammography. Our experience with a multi-modality screening program tailored to the needs of these specific high-risk individuals are reviewed in this article. Our data suggest that the prospective use of breast MR imaging offers a substantial increase in tumor detection rates compared to the combined use of two view mammography and state-of-the-art breast US. Interestingly, the considerable improvement in diagnostic sensitivity as offered by breast MRI was not traded off by a reduced specificity. On the contrary, MRI helped avoid many unnecessary biopsies that have been caused by false-positive high-frequency breast ultrasound findings. PMID- 12585664 TI - The UK national study of magnetic resonance imaging as a method of screening for breast cancer (MARIBS). AB - The UK national study of magnetic resonance imaging as a method of screening for breast cancer (MARIBS) is in progress. The study design, accrual to date, and related research projects are described. Revised accrual rates and expected recruitment are given. 15 cancers have been detected to date, from a total of 1236 screening measurements. This event rate and the tumour grades reported are compared with recent reports from other studies in women at high risk of breast cancer. PMID- 12585666 TI - BMRI in early detection of breast cancer in patients with increased genetic risk: our preliminary results. AB - Because of the higher risk of developing breast cancer and the early onset of the disease in women proved or suspected to be carriers of a breast cancer susceptibility gene, a dedicated screening should be offered as a less invasive approach with respect to the otherwise suggested prophylactic mastectomy. This should be optimized in order to overcome the limitations of conventional breast imaging with the application of new technologies such as Breast Magnetic Resonance Imaging (BMRI). A diagnostic protocol for routine control in patients with high risk for developing breast cancer has been prepared. Within a 7 months period, 23 patients suspected or proved to carry a breast cancer susceptibility gene underwent BMRI. Four breast cancers were identified with BMRI. In these cases mammography was negative because of the density of the parenchyma or for its fibroglandular pattern. US was negative in two cases, not specific for malignancy in one case and considered as only possibly malignant but with biopsy recommendation on the basis of MR findings in the last one. Clinic analysis was positive for mass in two cases. The accuracy of BMRI is known to be higher than that of conventional imaging in the study of breast parenchyma. High spatial resolution and no breast density influence can give more detailed information about smaller lesions and the right extent of the disease. PMID- 12585665 TI - The Italian multi-centre project on evaluation of MRI and other imaging modalities in early detection of breast cancer in subjects at high genetic risk. AB - This report presents the preliminary results of the first phase (21 months) of a multi-centre, non-randomised, prospective study, aimed at evaluating the effectiveness of contrast-enhanced magnetic resonance imaging (MRI), X-ray mammography (XM) and ultrasound (US) in early diagnosis of breast cancer (BC) in subjects at high genetic risk. This Italian national trial (coordinated by the Istituto Superiore di Sanita, Rome) so far recruited 105 women (mean age 46.0 years; median age 51.0; age range 25-77 years), who were either proven BRCA1 or BRCA2 mutation carriers or had a 1 in 2 probability of being carriers (40/105 with a previous personal history of BC). Eight cases of breast carcinomas were detected in the trial (mean age 55.3 years, median age 52.5; age range 35-70 years; five with previous personal history of BC). All trial-detected BC cases (8/8) were identified by MRI, while XM and US correctly classified only one. MRI had one false positive case, XM and US none. Seven "MRI-only" detected cancers (4 invasive, 3 in situ) occurred in both pre- (n = 2) and post-menopausal (n = 5) women. With respect to the current XM screening programmes addressed to women in the age range 50-69 years, the global incidence of BC in the trial (7.6%) was over ten-fold higher. The cost per "MRI-only" detected cancer in this particular category of subjects at high genetic risk was substantially lower than that of an XM-detected cancer in the general women population. These preliminary results confirmed that MRI is a very useful tool to screen subjects at high genetic risk for breast carcinoma, not only in pre-, but also in post-menopausal age, with a low probability of false positive cases. PMID- 12585667 TI - Usefulness of breast MRI in a patient with genetic risk. AB - We describe an interesting case-report represented by a patient carrying BRCA1 mutation, recruited for the study "Multicenter evaluation of Magnetic Resonance Imaging (MRI) in early diagnosis and prevention of breast cancer in high risk population", diagnosed with breast cancer on the basis of MRI findings but not with conventional mammography and ultrasound (US). She was already affected at 53 years of age by a multifocal Ductal Infiltrating Carcinoma (DIC) in the left breast; then, she had an axillary and sovraclavear nodal recurrence of the disease, three years after the initial diagnosis. Since other relatives were affected by breast cancer (mother, sister and niece) and two arose at early age (< 40 years), BRCA1 mutational analysis was offered to the patient, identifying a nonsense mutation on the exon 13. Furthermore, this patient was recruited to study contralateral breast and at the second round, two little foci, suspicious of malignancy, were identified only with MRI, but not with mammography and ultrasonography. The final diagnosis was multifocal Ductal Carcinoma in situ (DCIS); the major focus measured 3 mm. In our patient MRI has shown a major sensitivity with respect to conventional radiology and US and has provided a very early diagnosis in this woman at genetic risk. PMID- 12585668 TI - Breast MR imaging screening in eight women proved or suspected to be carriers of BRCA1&2 gene mutations. AB - The aim of this study was to evaluate the efficacy of MR imaging for the early detection of breast tumor in women at high genetic risk compared to conventional strategies such as ultrasonography and mammography. This study included 8 women, 5 of which had undergone surgery for breast cancer. BRCA germ line mutations were detected in 7 women, one patient was enrolled for more than 50% probability to be carrier of BRCA mutation. RM imaging screening was negative in 7 patients and strongly indicative of a malignant lesion in one. The gold standard was surgery for the suspicious cases and follow-up with clinical examination and conventional imaging every six months for the others. MR imaging proved itself to be a reliable technique in familial breast cancer high risk women. PMID- 12585669 TI - Observations on dental prostheses and restorations subjected to high temperatures: experimental studies to aid identification processes. AB - In large scale disasters associated with fire the damage caused by heat can make medico-legal identification of human remains difficult. Teeth, restorations and prostheses, all of which are resistant to even quite high temperatures can be used as aids in the identification process. In this project the behaviour and morphology of teeth and dental prostheses exposed to a range of high temperatures was studied. Healthy teeth, dental restorations and prostheses were placed in a furnace and heated at a rate of 30 degrees C/min and the effects of the predetermined temperatures 200, 400, 600, 800, 1000 and 1100 degrees C were examined by stereomicroscopy and scanning electron microscopy (SEM). Our observations show that some prostheses and restorative materials resist higher temperatures than theoretically predictable and that even when a restoration is lost because of detachment or change of state its ante-mortem presence can be confirmed and detected by both stereomicroscopic examination and SEM of the residual cavity. We further conclude that a reasonably reliable estimation of the temperature of exposure can be made from an analysis of the teeth and restorative materials. PMID- 12585670 TI - Fraudulent use of radiographic images. AB - The aim of this study was to evaluate the ability of trained observers to identify altered radiographic images after modifications using an image-editing software. Based on implantology in 10 radiographs, eight panoramic and one linear tomograph were modified while one tomograph was untouched. Implants were placed or removed and bone levels were altered, and seventy dentists were invited to identify these alterations. The results showed that the percentage of the correct answers was 12.5% or 2 identifications per examiner. The rate of false positives in relation to correct answers was at a level of 6:1. We concluded that the professionals have difficulty in identifying altered radiographs after using an image-editing software and that the seriousness of this situation demands that dentists be warned of the dangers of the use and abuse of this technology. PMID- 12585671 TI - Weighing evidence: quantitative measures of the importance of bitemark evidence. AB - Quantitative measures of the importance of evidence such as the "likelihood ratio" have become increasingly popular in the courtroom. These measures have been used by expert witnesses formally to describe their certainty about a piece of evidence. These measures are commonly interpreted as the amount by which the evidence should revise the opinion of guilt, and thereby summarize the importance of a particular piece of evidence. Unlike DNA evidence, quantitative measures have not been widely used by forensic dentists to describe their certainty when testifying about bitemark evidence. There is, however, no inherent reason why they should not be used to evaluate bitemarks. The purpose of this paper is to describe the likelihood ratio as it might be applied to bitemark evidence. We use a simple bitemark example to define the likelihood ratio, its application, and interpretation. In particular we describe how the jury interprets the likelihood ratio from a Bayesian perspective when evaluating the impact of the evidence on the odds that the accused is guilty. We describe how the dentist would calculate the likelihood ratio based on frequentist interpretations. We also illustrate some of the limitations of the likelihood ratio, and show how those limitations apply to bitemark evidence. We conclude that the quality of bitemark evidence cannot be adequately summarized by the likelihood ratio, and argue that its application in this setting may be more misleading than helpful. PMID- 12585672 TI - A critique of age estimation using attrition as the sole indicator. AB - The age determination of skeletal remains has been carried out using anthropological examination of the remaining bones and dentition. The aging of the dentition is based on attrition which, if physiological will correlate with age. Occasionally the only material available is a single tooth or a few teeth, or in the case of a living person, teeth in situ. In certain cases microscopic examination of the teeth may not be possible and the age estimation is then often determined by the degree of attrition associated with the tooth. In more recent times the causes of attrition have involved other factors such as bruxism, diet, environment and medication. The weaknesses and limitations of age estimation by examination of dental attrition as the sole indicator of age are highlighted. PMID- 12585673 TI - The current status of lip prints and their use for identification. AB - The use of lip prints for human identification was first suggested in 1950 and research was carried out on lip prints in the 1960s and early 1970s, resuming in the last few years. Although lip print identification has been utilized in court in isolated cases more research needs to be conducted in this field with regard to confirmation of uniqueness, and the collection and interpretation of evidence. Lip print identification needs to be acceptable in court as scientifically evidence based. PMID- 12585674 TI - The future of Latin American science. PMID- 12585675 TI - Neurosciences in Brazil: Rio de Janeiro. PMID- 12585677 TI - In vivo approach to the cellular mechanisms for sensory processing in sleep and wakefulness. AB - 1. The present review analyzes sensory processing during sleep and wakefulness from a single neuronal viewpoint. Our premises are that processing changes throughout the sleep-wakefulness cycle may be at least partially evidenced in single neurons by (a) changes in the phase locking of the response to the hippocampal theta rhythm, (b) changes in the discharge rate and firing pattern of the response to sound, and (c) changes in the effects of the neurotransmitters involved in the afferent and efferent pathways. 2. The first part of our report is based on the hypothesis that the encoding of sensory information needs a timer in order to be processed and stored, and that the hippocampal theta rhythm could contribute to the temporal organization. We have demonstrated that the guinea pig's auditory and visual neuronal discharge exhibits a temporal relationship (phase locking) to the hippocampal theta waves during wakefulness and sleep phases. 3. The concept that the neural network organization during sleep versus wakefulness is different and can be modulated by sensory signals and vice versa, and that the sensory input may be influenced by the CNS state, i.e., asleep or awake, is introduced. During sleep the evoked firing of auditory units increases, decreases, or remains similar to that observed during quiet wakefulness. However, there has been no auditory unit yet that stops firing as the guinea pig enters sleep. Approximately half of the cortical neurons studied did not change firing rate when passing into sleep while others increased or decreased. Thus, the system is continuously aware of the environment. We postulate that those neurons that changed their evoked firing during sleep are also related to still unknown sleep processes. 4. Excitatory amino acid neurotransmitters participate in the synaptic transmission of the afferent and efferent pathways in the auditory system. In the inferior colliculus, however, the effects of glutamate's mediating the response to sound and the efferent excitation evoked by cortical stimulation failed to show differences in sleep and wakefulness. 5. Considering that neonates and also infants spend most of the time asleep, the continuous arrival of sensory information to the brain during both sleep phases may serve to "sculpt" the brain by activity-dependent mechanisms of neural development, as has been postulated for wakefulness. PMID- 12585676 TI - Sexual differentiation of the brain: genes, estrogen, and neurotrophic factors. AB - Based on evidence obtained during the past 50 years, the current hypothesis to explain the sexual dimorphism of structure and function in the brain of vertebrates maintains that these differences are produced by the epigenetic action of gonadal hormones. However, evidence has progressively accumulated suggesting that genetic mechanisms controlling sexual-specific neuronal characteristics precede, or occur in parallel with, hormonal effects. 1. In cultures of hypothalamic neurons taken from gestation day 16 (GD16) embryos, treatment of sexually segregated cultures with estradiol (E2) induces axon growth in neurons from male neurons, but not from female neurons. In these cultures treatment with E2 increased the levels of tyrosine kinase type B (TrkB) and insulin-like growth factor I (IGF-I) receptors in male but not in female neurons. This and other sex differences cannot be explained by differences in hormonal environment, because the donor embryos were obtained when gonadal secretion of steroids is just beginning, before the perinatal surge of testosterone that determines development of the male brain beginning at GD17/18. 2. The response to estrogen is contingent upon coculture with heterotopic glia (mostly astrocytes) from a target region (amygdala) harvested from same-sex fetuses at GD16, whereas in the presence of homotopic glia or in cultures without glia, E2 had no effect. It was concluded that the axogenic effect of E2 depends on interaction between neurons and glia from a target region and that neurons from fetal male donors appear to mature earlier than neurons from females, a differentiated response that takes place prior to divergent exposure to gonadal secretions. 3. The effects of target and nontarget glia-conditioned media (CM) on the E2-induced growth of neuronal processes of hypothalamic neurons obtained from sexually segregated fetal donors were also studied. Estrogen added to media conditioned by target glia modified the number of primary neurites and the growth of axons of hypothalamic neurons of males but not of females. 4. Neither the Type III steroidal receptor blocker tamoxifen nor Type I antiestrogen ICI 182,780 prevented the axogenic effects of the hormone. Estradiol made membrane impermeable by conjugation to a protein of high molecular weight (E2-BSA) preserved its axogenic capacity, suggesting the possibility of a membrane effect responsible for the action of E2. 5. Western blot analysis of the tyrosine kinase type A (TrkA), type B (TrkB), type C (TrkC), and insulin-like growth factor (IGF I R) receptors in extracts from homogenates of cultured hypothalamic neurons showed that in cultures of male-derived neurons grown with E2 and CM from target glia, the amounts of TrkB and IGF-I R increased notably. Densitometric quantification showed that these cultures had more TrkB than cultures with CM alone or E2 alone. On the contrary, in cultures of female-derived neurons, the presence of CM alone induced maximal levels of TrkB, which were not further increased by E2; female-derived neurons in all conditions did not contain IGF-I R. Levels of TrkC were not modified by any experimental condition in male- or female-derived cultures and Trk A was not found in the homogenates. These results are compared with similar data from other laboratories and integrated in a model for the confluent interaction of estrogen and neurotrophic factors released by glia that may contribute to the sexual differentiation of the brain. PMID- 12585678 TI - Thyroid hormone actions on neural cells. AB - 1. In addition to its role in cellular metabolic activity, thyroid hormone (TH) is critically involved in growth, development, and function of the central nervous system. In the brain, as in other structures, TH is described to exert its major action by the binding of L-3,5,3'-triiodothyronine (T3), considered as the bioactive form of the hormone, to nuclear thyroid hormone receptors (TR) that function as ligand-dependent transcription factors. 2. The transcription of numerous brain genes was indeed shown to be positively or negatively regulated by TH, turning these TR-mediated effects one explanation for the physiological effects of TH. In this context, the knowledge from TR-knockout studies provides some surprising results, since neonatal hypothyroidism is associated to more significant abnormalities than is TR deficiency. Some (nonexclusive) hypotheses include a permissive effect of TH, allowing derepression of unliganded-TR effects and non-TR-mediated effects of the hormone, further emphasizing the importance of a controlled accessibility of neural cells to TH. 3. On the other hand, T3 was demonstrated to directly act not only on neuronal but also on glial cells proliferation and differentiation, contributing to the harmonious development of the brain. Interestingly, in addition to these direct actions on neuronal and glial cells, several lines of evidence, notably developped in our laboratory, point out the role of thyroid hormone in neuronal-glial interactions. PMID- 12585679 TI - Beta-amyloid production, aggregation, and clearance as targets for therapy in Alzheimer's disease. AB - 1. Despite major efforts aimed at elucidating the molecular basis and physiopathology of Alzheimer's disease (AD), there is still no effective treatment available for this devastating disorder. The biological mechanisms underlying the development of AD are complex, as multiple factors appear to modulate (either positively or negatively) the progression of neurodegeneration in the brains of AD patients. Not surprisingly, a number of different therapeutic approaches aimed at distinct aspects of the disease are currently being pursued. Given its central role in the neuropathology of AD, the beta-amyloid peptide (Abeta) is the focus of many such approaches. 2. In this review, we discuss recent developments along three major lines of investigation: (i) identification and characterization of inhibitors of the enzymes involved in proteolytic processing of the amyloid precursor protein and production of Abeta; (ii) identification of the pathways involved in cerebral degradation and clearance of Abeta; and (iii) characterization of small-molecule inhibitors of amyloid aggregation that prevent cerebral amyloid deposition and neurotoxicity. 3. Significant progress has been achieved in these directions, opening up new perspectives toward the development of effective approaches for the treatment or prevention of AD. PMID- 12585681 TI - Phoneutria nigriventer venom: a cocktail of toxins that affect ion channels. AB - 1. We review the pharmacological actions of toxins present in the venom of the aggressive spider Phoneutria nigriventer. 2. This venom is rich in toxins that affect ion channels and neurotransmitter release. Voltage-gated sodium, calcium, and potassium channels have been described as the main targets of these toxins. 3. In addition to these classical actions Phoneutria toxins have also been shown to affect glutamate transporter. 4. It is expected that molecular genetics in addition to biochemical, biophysical and pharmacological approaches will help to further define Phoneutria toxins and their mechanisms of action in the near future. PMID- 12585680 TI - Signals involved in targeting membrane proteins to synaptic vesicles. AB - 1. Synaptic vesicles (SVs) mediate fast regulated secretion of classical neurotransmitters. In order to perform their task SVs rely on a restrict set of membrane proteins. The mechanisms responsible for targeting these proteins to the SV membrane are still poorly understood. 2. Likewise, little is known about the intracellular routes taken by these proteins in their way to SV membrane. Recently, several domains and motifs necessary for correct localization of SV proteins have been identified. 3. In this review we summarize the sequence motifs that have been identified in the cytoplasmic domains of SV proteins that are involved in endocytosis and targeting of SVs. We suggest that the vesicular acetylcholine transporter, a protein found predominantly in synaptic vesicles, is perhaps a model protein to understand the pathways and interactions that are used for synaptic vesicle targeting. PMID- 12585682 TI - Brain oscillations, medium spiny neurons, and dopamine. AB - 1. The striatum is part of a multisynaptic loop involved in translating higher order cognitive activity into action. The main striatal computational unit is the medium spiny neuron, which integrates inputs arriving from widely distributed cortical neurons and provides the sole striatal output. 2. The membrane potential of medium spiny neurons' displays shifts between a very negative resting state (down state) and depolarizing plateaus (up states) which are driven by the excitatory cortical inputs. 3. Because striatal spiny neurons fire action potentials only during the up state, these plateau depolarizations are perceived as enabling events that allow information processing through cerebral cortex basal ganglia circuits. In vivo intracellular recording techniques allow to investigate simultaneously the subthreshold behavior of the medium spiny neuron membrane potential (which is a "reading" of distributed patterns of cortical activity) and medium spiny neuron firing (which is an index of striatal output). 4. Recent studies combining intracellular recordings of striatal neurons with field potential recordings of the cerebral cortex illustrate how the analysis of the input-output transformations performed by medium spiny neurons may help to unveil some aspects of information processing in cerebral cortex-basal ganglia circuits, and to understand the origin of the clinical manifestations of Parkinson's disease and other neurologic and neuropsychiatric disorders that result from alterations in dopamine-dependent information processing in the cerebral cortex-basal ganglia circuits. PMID- 12585683 TI - Considerations on the astroglial architecture and the columnar organization of the cerebral cortex. AB - Present knowledge on astroglial roles in brain organization and function indicates that these cells can regulate the extracellular ionic composition and modulate neuronal activity. In this regard, the "panglial synctium" formed by electrically and chemically coupled stellate astrocytes ("general mammalian" architecture) is believed to provide an intracellular pathway for the redistribution of ions and molecules within the cerebral cortex. Long astroglial interlaminar processes ("primate-specific" architecture) that run parallel to apical dendrites in the cerebral cortex of primates, may provide additional properties to the glial participation in cortical physiology and function. Since these processes are exclusively present within the primate order, functional models of cortical computations for these species should incorporate the astroglial interlaminar architecture in addition to the panglial synctium. This study analyzes possible implications of interlaminar astroglial processes, for the regulation of the extracellular ionic composition and segregation of functional columns in the cerebral cortex. PMID- 12585684 TI - Basal ganglia and functions of the autonomic nervous system. AB - 1. The aim of this mini-review was to describe an underrecognized but important aspect of the basal ganglia diseases, the dysfunction of the autonomic nervous system that patients suffer owing to the degenerative process affecting these structures, mainly Parkinson's disease. 2. We analyze the most prevalent autonomic abnormalities in these patients from an experimental and clinical point of view. PMID- 12585685 TI - Opioidergic effects of nonopioid analgesics on the central nervous system. AB - 1. The analgesic effect of nonsteroidal anti-inflammatory drugs (NSAIDs) is partly due to the fact that they act upon the periaqueductal gray matter (PAG) and the rostral ventromedial medulla of the brain stem and thus activate the descending pain-control system, which inhibits nociceptive transmission at the spinal dorsal horn. 2. The analgesic action of dipyrone (metamizol) and of lysine acetylsalicylate (LASA), two well-known NSAIDs. whether microinjected into the PAG or given systemically, can be reverted by naloxone. Repeated administration of dipyrone or LASA induces tolerance to their antinociceptive effect, with cross tolerance to morphine, and a withdrawal syndrome upon naloxone administration. Dipyrone tolerance can be reverted by proglumide, a cholecystokinin antagonist. 3. These findings reveal a close association between the central action of NSAIDs and endogenous opioids. PMID- 12585686 TI - Signaling mechanisms mediating BDNF modulation of memory formation in vivo in the hippocampus. AB - Given that brain-derived neutrophic factor (BDNF) modulates both short-term synaptic function and activity-dependent synaptic plasticity in the adult hippocampus, here we examined signaling mechanisms in vivo in the hippocampus mediating BDNF modulation of long-term memory (LTM) formation of a one-trial fear motivated learning task in rats. Bilateral infusions of function-blocking anti BDNF antibody into the CA1 region of the dorsal hippocampus decreased extracellular-signal regulated kinase 2 (ERK2) and CREB activation and impaired LTM retention scores. Inhibition of ERK1/2 activation by PD098059 produced similar effects and also reduced CREB phosphorylation. In contrast, intrahippocampal administration of recombinant human BDNF increased ERK1/2 and CREB activation and facilitated LTM. Activated-p38, activated-PKC isoforms, and activated-AKT were unaltered after BDNF or anti-BDNF infusion. In addition, no changes were found on alphaPKA and betaPKA catalytic subunits in nuclear samples. Thus, our results suggest that BDNF exerts its role in LTM formation in vivo in CA1 region of the hippocampus, at least in part, via CREB activation. Moreover, BDNF-induced CREB activation appears to be mediated mainly through the activation of ERK1/2 signaling pathway. PMID- 12585687 TI - Involvement of 5-HT(2A/2B/2C) receptors on memory formation: simple agonism, antagonism, or inverse agonism? AB - 1. The 5-HT2 receptors subdivision into the 5-HT(2A/2B/2C) subtypes along with the advent of the selective antagonists has allowed a more detailed investigation on the role and therapeutic significance of these subtypes in cognitive functions. The present study further analyzed the 5-HT2 receptors role on memory consolidation. 2. The SB-200646 (a selective 5-HT(2B/2C) receptor antagonist) and LY215840 (a nonselective 5-HT(2/7) receptor antagonist) posttraining administration had no effect on an autoshaped memory consolidation. However, both drugs significantly and differentially antagonized the memory impairments induced by 1-(3-chlorophenyl)piperazine (mCPP), 1-naphtyl-piperazine (1-NP), mesulergine, or N-(3-trifluoromethylphenyl) piperazine (TFMPP). 3. In contrast, SB-200646 failed to modify the facilitatory procognitive effect produced by (+/-)-2.5 dimethoxy-4-iodoamphetamine (DOI) or ketanserin, which were sensitive to MDL100907 (a selective 5-HT2A receptor antagonist) and to a LY215840 high dose. 4. Finally, SB-200646 reversed the learning deficit induced by dizocilpine, but not that by scopolamine: while SB-200646 and MDL100907 coadministration reversed memory deficits induced by both drugs. 5. It is suggested that 5-HT(2B/2C) receptors might be involved on memory formation probably mediating a suppressive or constraining action. Whether the drug-induced memory impairments in this study are explained by simple agonism, antagonism, or inverse agonism at 5-HT2 receptors remains unclear at this time. 6. Notably, the 5-HT2 receptor subtypes blockade may provide some benefit to reverse poor memory consolidation conditions associated with decreasedcholinergic, glutamatergic, and/or serotonergic neurotransmission. PMID- 12585688 TI - Glutamate NMDA receptor subunit R1 and GAD mRNA expression in human temporal lobe epilepsy. AB - 1. Molecular mechanisms underlying increased hippocampal excitability in human temporal lobe epilepsy (TLE) are largely unknown. A disturbance of the imbalance between excitatory and inhibitory neurotransmission pathways in the epileptic hippocampus may contribute substantially to a decreased seizure threshold. 2. We have extended the investigation whether TLE is associated with changes in the expression of GAD67 and NMDAR1 by assessing the relative amounts of the mRNAs in human hippocampal samples by means of semiquantitative RT-PCR. The samples included 16 hippocampal slices obtained at surgery from intractable TLE (HS, n = 14; non-HS, n = 2) and 3 postmortem control hippocampi. 3. The ratio for the GAD/NMDAR1 transcripts was significantly higher in TLE cases when compared to the nonepileptic samples. Such findings are mainly a consequence of the increased amounts of GAD mRNA detected in the epileptic hippocampus. Compared with nonepileptic samples, and without correction for neuron losses, the amounts of NMDAR1 mRNA in HS are slightly reduced, and in the non-HS samples they are significantly increased, which is consistent with an increase of NMDAR1 in the hippocampal remaining neurons, as previously reported. 4. Our results also contribute to the indication of GAD67 mRNA upregulation in human TLE. A possible functional implication for the increased GAD mRNA levels could be a mechanism to reduce neuronal hyperexcitability, synchronization, and/or the spread of seizure. PMID- 12585689 TI - Effect of melatonin on changes in locomotor activity rhythm of Syrian hamsters injected with beta amyloid peptide 25-35 in the suprachiasmatic nuclei. AB - 1. Alzheimer's disease is associated with circadian rhythm disturbances, probably because of beta amyloid-induced neuronal damage of hypothalamic suprachiasmatic nuclei (SCN). 2. Since there is no published study on the circadian consequences of injecting beta amyloid peptide in experimental animals, one objective of the present study was to examine circadian locomotor activity in Syrian hamsters injected with beta amyloid peptide 25-35 into both SCN. 3. Because one of the proposed therapies for circadian alterations in dementia is the administration of melatonin, a chronobiotic agent with antioxidant properties, the preventive effect of melatonin on the circadian changes produced by beta amyloid microinjection into SCN was also assessed. 4. Wheel running activity was recorded by using the Dataquest III system in male golden hamsters kept under 14:10 light dark photoperiods. Animals received microinjections of beta amyloid peptide 25-35 (100 microM solution, 1 microL) or saline in each SCN. Only those animals with neuronal lesions larger than 10% of SCN after beta amyloid injection were considered for further analysis. 5. To assess the effect of melatonin on beta amyloid peptide activity, melatonin was given in the drinking water (25 microg/mL) starting 15 days in advance to the microinjection of beta amyloid peptide into SCN. 6. Beta amyloid-treated hamsters exhibited a significant phase advance of onset of running activity of about 22 min as compared to saline injected animals. They also showed a significantly greater variability in onset time of wheel running activity, mainly evident from 6 to 15 days of treatment. 7. Melatonin administration in the drinking water prevented the phase advance of onset time and the increased variability of onset time brought about by beta amyloid peptide. 8. The results support the existence of a neuroprotective effect of melatonin on beta amyloid-induced circadian changes in hamsters. PMID- 12585690 TI - Cytoplasmic c-Jun N-terminal immunoreactivity: a hallmark of retinal apoptosis. AB - 1. We investigated the association of c-Jun with apoptosis within retinal tissue. Explants of the retina of neonatal rats were subject to a variety of procedures that cause apoptosis of specific classes of retinal cells at distinct stages of differentiation. The expression of c-Jun was detected by Western Blot, and immunohistochemistry was done with antibodies made for either N-terminal or C terminal domains of c-Jun, and correlated with apoptosis detected either by chromatin condensation or by in situ nick end labeling of fragmented DNA. 2. c Jun protein content was increased in retinal tissue subject to induction of both photoreceptor and ganglion cell death. 3. c-Jun N-terminal immunoreactivity was found mainly in the cytoplasm of apoptotic cells regardless of cell type, of the stage of differentiation, including proliferating cells, or of the means of induction of apoptosis. 4. The data are consistent with the hypothesis that c-Jun is involved in the control of cell death in retinal tissue, but other proteins that cross-react with c-Jun N-terminal antibodies may also be major markers of retinal apoptosis. 5. Antibodies directed to c-Jun N-terminal (aa 91-105) are useful tools to follow apoptotic changes in retinal tissue. PMID- 12585691 TI - Regulation of vesicular acetylcholine transporter by the activation of excitatory amino acid receptors in the avian retina. AB - 1. Previous studies have shown that phorbol esters induce protein kinase C (PKC) mediated phosphorylation of the vesicular acetylcholine transporter (VAChT) and change its interaction with vesamicol. However, it is not clear whether physiological activation of receptors coupled to PKC activation can alter VAChT behavior. 2. Here we tested whether activation of kaianate (KA) receptors alters VAChT. Several studies suggest that the cholinergic amacrine cells display KA/AMPA receptors that mediate excitatory input to these neurons. In addition, KA in the chicken retina can generate intracellular messengers with the potential to regulate cellular functions. 3. In cultured chicken retina (E8C11) KA reduced vesamicol binding to VAChT by 53%. This effect was potentiated by okadaic acid, a protein phosphatase inhibitor, and was totally prevented by BIM, a PKC inhibitor. 4. Phorbol myristate acetate (PMA), but not alpha-PMA, reduced in more than 85% the number of L-[3H]-vesamicol-specific binding sites in chicken retina, confirming that activation of PKC can influence vesamicol binding to chicken VAChT. 5. The data show that activation of glutamatergic receptors reduces [3H] vesamicol binding sites (VAChT) likely by activating PKC and increasing the phosphorylation of the ACh carrier. PMID- 12585692 TI - Kainic acid modifies mu-receptor binding in young, adult, and elderly rat brain. AB - Mu-receptor binding changes were evaluated following the kainic acid (KA)-induced status epilepticus (SE) in young, adult, and elderly animals. Male Wistar rats were used as follows: young rats (15 days old) were treated with KA (7 mg/kg) and sacrificed 72 h (YKA3d) or 35 days (YKA35d) after SE; adult (90 days old) (AKA1d and AKA40d) and elderly rats (1-year-old) (EKA1d and EKA40d) were injected with KA (10 mg/kg) and then sacrificed 24 h or 40 days following SE. Their brains were processed for an autoradiography assay for mu-receptors. The YKA3d group showed increased values in dentate gyrus (39%) and a decrease in substantia nigra (26%); YKA35d animals had a reduction in caudate putamen (29%) and in substantia nigra (20%). The AKA1d group exhibited increased mu-receptors in caudate putamen (49%), cingulate (415%), frontal (52%), and temporal (53%) cortices: substantia nigra (56%), dentate gyrus (48%). and CA2 field of hippocampus (53%). The AKA40d group showed increased values in sensorimotor cortex (45%), anterior (39%), medial (65%), basolateral (202%), and central (32%) amygdaloid nuclei; dentate gyrus (80%) as well as CA2 (80%) and CA3 (49%) fields of hippocampus. The EKA1d group presented decreased mu-receptor binding in piriform (16%) and enthorinal (22%) cortices as well as in anterior amygdala nucleus (17%). The EKA40d group showed reduced values in sensorimotor cortex (14%) and substantia nigra (27%). The present results indicate that the mu-binding changes following SE depend on the rate of brain maturation. PMID- 12585693 TI - Bile secretion is centrally regulated by C-type natriuretic peptide. AB - 1. Current evidence supports that C-type natriuretic peptide (CNP) is the brain natriuretic peptide. Natriuretic peptide receptors and mRNA CNP have been reported in the liver and in discrete areas and nucleus of the central nervous system involved in the regulation of gastrointestinal physiology. In the present work, we sought to establish the role of CNP in the central regulation of bile secretion in the rat and to delineate the possible pathways and mechanisms involved. 2. To examine the role of CNP on bile secretion, the peptide was applied in the brain lateral ventricle (1, 10, and 100 ng/microL) and bile samples were collected every 15 min for 60 min. The role of the autonomic nervous system in CNP response was assessed by atropine or combined phentolamine and propranolol administration. 3. Centrally applied CNP diminished basal as well as bile salt-evoked bile flow in a dose-dependent manner. CNP reduced bile acid output as well as sodium and potassium excretion, supporting CNP effect on bile acid-dependent flow. CNP also decreased chloride excretion and increased bile pH. The excretion of total glutathione was not affected by centrally applied CNP suggesting that this peptide does not alter bile acid-independent flow. Neither parasympathetic nor sympathetic blockade abolished CNP inhibitory response on bile secretion. Mean arterial pressure and portal venous pressure were not modified by CNP. 4. Present findings show that centrally applied CNP modulates bile secretion in a dose-dependent fashion. CNP alkalinized bile and reduced bile acid-dependent flow without affecting bile acid-independent flow. The inhibitory response of CNP on bile secretion was not mediated by the autonomic nervous system. Present findings give further support to the role of CNP as the brain natriuretic peptide. PMID- 12585694 TI - Atrial natriuretic factor (ANF) effects on L-, N-, and P/Q-type voltage-operated calcium channels. AB - 1. We have previously reported that atrial natriuretic factor (ANF) decreases neuronal norepinephrine (NE) release. The mechanism that mediates NE release from presynaptic membrane to synaptic cleft is a strongly calcium-dependent process. The modulator effect of ANF may be related to modifications in calcium influx at the presynaptic nerve ending by interaction with voltage-operated calcium channels (VOCCs). 2. On this basis we investigated the effects of ANF on K+ induced 45Ca2+ uptake and evoked neuronal NE release in the presence of specific L-, N-, and P/Q-type calcium channel blockers in the rat hypothalamus. 3. Results showed that ANF inhibited K+-induced 45Ca2+ uptake in a concentration-dependent fashion. Concentration-response curves to VOCC blockers nifedipine (NFD, L-type channel blocker), omega-conotoxin GVIA (CTX, N-type channel blocker), and omega agatoxin IVA (AGA, P/Q-type channel blocker) showed that all the blockers decreased NE release. Incubation of ANF plus NFD showed an additive effect as compared to NFD or ANF alone. However, when the hypothalamic tissue was incubated in the presence of ANF plus CTX or AGA there were no differences in neuronal NE release as compared to calcium channel blockers or ANF alone. 4. These results suggest that ANF decreases NE release by an L-type calcium channel independent mechanism by inhibiting N- and/or P/Q-type calcium channels at the neuronal presynaptic level. Thus, ANF modulates neuronal NE release through different mechanisms involving presynaptic calcium channel inhibition. PMID- 12585695 TI - Role of endothelin type B receptor in NO/cGMP signaling pathway in rat median eminence. AB - We studied the effect of endothelins (ETs) on receptor-mediated NO/cGMP signaling in rat arcuate nucleus-median eminence (AN-ME) fragments, an hypothalamic structure known to contain a rich plexus of nitric oxide synthase (NOS) containing neurons and fibers together with densely arranged ET(B)-receptor-like immunoreactive fibers. NOS activity was determined measuring the conversion of [3H] arginine to [3H] citrulline, as an index of NO produced. cGMP production was determined by radio immunoassay. ET-1, ET-3, and the selective ET(B) receptor agonist, IRL1620, significantly increased cGMP formation and NOS activity. Preincubation of AN-ME fragment with L-arginine analog, N-nitro-L-arginine (L NAME), inhibited ET-1 or IRL1620-stimulated cGMP formation. The addition of theselective ET(B) receptor antagonist, BQ788, blocked ET-1-, ET-3-, or IRL1620 induced increase in NOS activity and cGMP generation, while BQ123, a selective ET(A) receptor antagonist, was ineffective. Our results demonstrate that in whole rat AN-ME fragments, ETs stimulate NO/cGMP signaling pathway through the interaction with the ET(B) receptor subtype, supporting the concept that ETs may represent an important regulator of reproductive and neuroendocrine function. PMID- 12585696 TI - Serotonin transporter modulation in blood lymphocytes from patients with major depression. AB - 1. Serotonin is a neurotransmitter in the central nervous system which has been implicated in the aetiology and pathogenesis of affective disorders. The serononergic system also plays several roles in the immune system through the expression of a number of its receptor subtypes in the immune cells. 2. Following release serotonin is inactivated by reuptake into neurons and other cells by a specific serotonin sodium and chloride-dependent transporter molecule, whose structure has been elucidated. 3. Measurement [3H]paroxetine binding showed that human lymphocytes contain a high-affinity serotonin transporter. 4. To assess the serotonin function in major depression, we investigated serotonin transporter density in blood lymphocytes from patients with this disorder and selected according to the interview of the American Psychiatric Association. 5. Patients were divided into two groups and treated with two different antidepressant drugs, one group receiving fluoxetine, a selective serotonin reuptake inhibitor, and another mirtazapine, an antagonist of alpha2-adrenergic auto and heteroreceptors, for a period of 6 weeks. 6. Blood samples were obtained before and after the treatment, lymphocytes were isolated by Ficoll/Hypaque gradient, subjected to differential adhesion to plastic, and cell membranes were prepared for binding assay of [3H]paroxetine. 7. Lymphocytes serotonin transporter number was significantly reduced, while the affinity was unchanged, in patients with major depression disorder as compare to controls. 8. In addition, there was a partial recovery in lymphocytes serotonin (5HT) transporter number in the period posterior to the antidepressants administration, accompanied with clinical and depression rating scales improvement. Serotonin was determined in platelet-poor plasma and in lymphocytes before and after drugs administration, showing a significant decrease in the patients treated compared to untreated and controls. 9. These results are evidence of the potential interaction between the nervous and immune systems. The mechanisms underlying this interaction are under study, and might be related to modifications in the expression or function of the serotonin transporters in lymphocytes of depressed patients. PMID- 12585697 TI - Depolarization-evoked GABA release from myenteric plexus is partially coupled to L-, N-, and P/Q-type calcium channels. AB - 1. There are many evidences suggesting that gamma-aminobutyrate (GABA) is an important neurotransmitter and/or neuromodulator in the gut. 2. Using the myenteric plexus-longitudinal muscle preparation from the guinea pig ileum, we investigated the evoked release of [3H] GABA from enteric neurons by electrical pulses or high KCl, which occurs in a calcium-dependent and -independent way. In addition, using selective calcium channel blockers, we report the participation of distinct subtypes of calcium channels in the evoked release, showing a minor participation of L- and Q-type calcium channels, while N- and P-type have a participation of approximately 15%, each. However, regardless of the combination of Ca2+ channel blockers, we did not observe an inhibition greater than 50% of the calcium-dependent component of [3H] GABA release. 3. Thus, while the observed Ca2+-independent release mostly probable occur via reversal of the membrane GABA transporter, in our conditions, a considerable portion of the Ca2+-dependent evoked release of [3H] GABA is not coupled to L-, N-, or P/Q-type calcium channels, suggesting the involvement of intracellular calcium stores or other ways of getting calcium across the membrane. PMID- 12585698 TI - Analysis of T102C 5HT2A polymorphism in Brazilian psychiatric inpatients: relationship with suicidal behavior. AB - 1. Central serotonergic dysfunction and genetic factors are associated with suicidal behavior in psychiatric patients. The goal of this study was to examine the association between the 5-HT2A gene polymorphism (102T/C) and suicide in a sample of Brazilian psychiatric inpatients. 2. We studied 225 subjects. Genotypic frequencies were obtained after DNA extraction and the region of 5-HT2A/T102C containing the polymorphic site amplified by the polymerase chain reaction and digested with the restriction enzyme HpaII. 3. No differences were found between patients with and without suicide attempt history. Patients with a history of severe suicide attempts also did not exhibit different genotypic frequencies when compared with patients without a suicide attempt history. 4. These results suggest that the 5HT2A gene polymorphism (102T/C) may not be involved in the genetic susceptibility to suicidal behavior. PMID- 12585699 TI - Modulation of Na+-channels by neurotoxins produces different effects on [3H]ACh release with mobilization of distinct Ca2+-channels. AB - 1. Voltage-gated Na+ channels are responsible for initiation and conduction of action potentials. The arrival of an action potential at nerve terminal increases intracellular Na+ and Ca2+ concentrations. Calcium entry into neurons through voltage-dependent calcium channels is associated with a variety of intracellular processes. Scorpion neurotoxins have been used as tools to investigate mechanisms involved in neurotransmitter release. Tityustoxin (TsTX) is an alpha-type toxin that delays Na+-channel inactivation. Toxin-gamma (TiTX-gamma) is a beta-type toxin that induces Na+-channel activation at resting potentials. 2. In the present work, we describe the effects of both toxins on [3H]acetylcholine ([3H]ACh) release from rat cerebrocortical synaptosomes, in the presence or absence of the calcium channels blockers: omega-conotoxin-GVIA (omega-CgTx), 1 microM; omega-agatoxin-IVA (omega-Aga), 30 nM; omega-conotoxin-MVIIC (omega MVIIC), 1 microM; or verapamil, 1 microM. 3. TsTX evokes [3H]ACh release in a concentration-dependent manner with a gradual increase up to saturation at concentrations of 500 nM. However, release of ACh evoked by TiTX-gamma was not linear regarding the toxin concentration. The [3H]-ACh release evoked by TsTX or TiTX-gamma was partially inhibited by omega-CgTx or omega-Aga, and blocked with omega-MVIIC. Verapamil (1 microM) had no effect. Tetrodotoxin blocked [3H]ACh release evoked by both toxins. 4. These results show that different actions on Na+-channels produce different effects on [3H]ACh release with involvement of distinct presynaptic Ca2+-channels, which supports the idea that sodium channels may modulate neurotransmitter release. PMID- 12585700 TI - Treatment with 7-nitroindazole enhances kainic acid induced cholinergic neurotoxicity in the rat striatum: a neuroprotective role for neuronal nitric oxide. AB - 1. In this study we investigated the effect of 7-nitroindazole (7-NI), a preferential inhibitor of neuronal nitric oxide synthase (nNOS), on kainic acid (KA) induced neurotoxicity in rats. Choline acetyltransferase activity (CAT), a cholinergic marker, and histological changes were employed to assess neurotoxicity. 2. In control rats, the local intrastriatal injection of 0.5 microg of KA reduced CAT from 22.9 +/- 2.2 to 14.7 +/- 2.0 nmol/h/mg tissue ((38 +/- 6)% reduction) (P < 0.001). Greater reductions in CAT were observed with 1 and 2 microg of KA ((70 +/- 6)% and (80 +/- 3)%, respectively). 7-NI aggravated KA-induced cholinergic and histological damage. KA reduced CAT by (68.2 +/- 4)% in 7-NI-treated rats, by (38 +/- 6)% in saline-treated controls, and by (41 +/- 4)% in peanut-oil- (7-NI-vehicle-) treated rats (P = 0.0047). 3. After KA, CAT activity averaged 14.3 +/- 2.0 in peanut-oil-treated rats and 7.9 +/- 1.0 nmol/h/mg tissue in 7-NI- (peanut-oil-) treated rats (P = 0.015). Similarly to changes in CAT, 7-NI treatment aggravated KA-induced histological changes indicative of neuronal damage (acute ischemic neuronal changes, disorganization of myelinated fibers bundle, and vacuolation changes of the neuropil). Treatment with 7-NI was not associated with increased mortality. 4. Our findings suggest that neuronal NO plays a neuroprotective action on excitotoxicity. PMID- 12585702 TI - Image processing and analysis in drug discovery and clinical trials. PMID- 12585701 TI - Lesion of caudate-putamen interneurons with kainic acid alters dopamine and serotonin metabolism in the olfactory tubercle of the rat. AB - 1. The existence of functional interrelationships between dorsal and ventral regions of the rat striatum was investigated. Kainic acid (KA) was employed to induce neuronal lesions in the more dorsal striatum, the caudate-putamen (CP). Only one CP (one side) received KA. KA-induced neurotoxicity at the site of injection (CP) was evidenced by reductions in choline-acetyltransferase activity and in GABA levels, and by increases in the ratios metabolite/monoamine for dopamine (DA) and serotonin (5-HT). 2. In addition to the well-known local effects, direct stereotaxic injection of KA into the CP produced distant effects in the ipsilateral olfactory tubercle (OT). A dose-dependent increase in the levels of 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5 hydroxyindoleacetic acid (5-HIAA) and decreases in DA and 5-HT concentrations were observed in the OT ipsilateral to the CP injected with KA. With 1, 2, 3, and 4 microg of KA, the ratio DOPAC+HVA/DA in the OT was 30, 79, 140, and 173% higher, respectively, than control levels. With 2, 3, and 4 microg of KA, the levels of 5-HIAA were approximately 30, 60, and 120% higher than control values, and the changes in 5-HIAA were associated with significant reductions in 5-HT concentrations. 3. Our results suggest that the dorsal part of the striatum exerts important regulatory functions over the most ventral striatal region, the OT. Destruction of CP interneurons by KA leads to disinhibition of DA and 5-HT activities to the OT. The functional interactions between dorsal and ventral striatal regions may play a role in the integration of fundamental life preserving, motivational, and goal-directed olfactory motor behaviors of rodents. PMID- 12585703 TI - Segmentation and tracking of migrating cells in videomicroscopy with parametric active contours: a tool for cell-based drug testing. AB - This paper presents a segmentation and tracking method for quantitative analysis of cell dynamics from in vitro videomicroscopy data. The method is based on parametric active contours and includes several adaptations that address important difficulties of cellular imaging, particularly the presence of low contrast boundary deformations known as pseudopods, and the occurence of multiple contacts between cells. First, we use an edge map based on the average intensity dispersion that takes advantage of relative background homogeneity to facilitate the detection of both pseudopods and interfaces between adjacent cells. Second, we introduce a repulsive interaction between contours that allows correct segmentation of objects in contact and overcomes the shortcomings of previously reported techniques to enforce contour separation. Our tracking technique was validated on a realistic data set by comparison with a manually defined ground truth and was successfully applied to study the motility of amoebae in a biological research project. PMID- 12585704 TI - Tracking leukocytes in vivo with shape and size constrained active contours. AB - Inflammatory disease is initiated by leukocytes (white blood cells) rolling along the inner surface lining of small blood vessels called postcapillary venules. Studying the number and velocity of rolling leukocytes is essential to understanding and successfully treating inflammatory diseases. Potential inhibitors of leukocyte recruitment can be screened by leukocyte rolling assays and successful inhibitors validated by intravital microscopy. In this paper, we present an active contour or snake-based technique to automatically track the movement of the leukocytes. The novelty of the proposed method lies in the energy functional that constrains the shape and size of the active contour. This paper introduces a significant enhancement over existing gradient-based snakes in the form of a modified gradient vector flow. Using the gradient vector flow, we can track leukocytes rolling at high speeds that are not amenable to tracking with the existing edge-based techniques. We also propose a new energy-based implicit sampling method of the points on the active contour that replaces the computationally expensive explicit method. To enhance the performance of this shape and size constrained snake model, we have coupled it with Kalman filter so that during coasting (when the leukocytes are completely occluded or obscured), the tracker may infer the location of the center of the leukocyte. Finally, we have compared the performance of the proposed snake tracker with that of the correlation and centroid-based trackers. The proposed snake tracker results in superior performance measures, such as reduced error in locating the leukocyte under tracking and improvements in the percentage of frames successfully tracked. For screening and drug validation, the tracker shows promise as an automated data collection tool. PMID- 12585705 TI - A contribution of image processing to the diagnosis of diabetic retinopathy- detection of exudates in color fundus images of the human retina. AB - In the framework of computer assisted diagnosis of diabetic retinopathy, a new algorithm for detection of exudates is presented and discussed. The presence of exudates within the macular region is a main hallmark of diabetic macular edema and allows its detection with a high sensitivity. Hence, detection of exudates is an important diagnostic task, in which computer assistance may play a major role. Exudates are found using their high grey level variation, and their contours are determined by means of morphological reconstruction techniques. The detection of the optic disc is indispensable for this approach. We detect the optic disc by means of morphological filtering techniques and the watershed transformation. The algorithm has been tested on a small image data base and compared with the performance of a human grader. As a result, we obtain a mean sensitivity of 92.8% and a mean predictive value of 92.4%. Robustness with respect to changes of the parameters of the algorithm has been evaluated. PMID- 12585706 TI - Digital stereo image analyzer for generating automated 3-D measures of optic disc deformation in glaucoma. AB - The major limitations of precise evaluation of retinal structures in present clinical situations are the lack of standardization, the inherent subjectivity involved in the interpretation of retinal images, and intra- as well as interobserver variability. While evaluating optic disc deformation in glaucoma, these limitations could be overcome by using advanced digital image analysis techniques to generate precise metrics from stereo optic disc image pairs. A digital stereovision system for visualizing the topography of the optic nerve head from stereo optic disc images is presented. We have developed an algorithm, combining power cepstrum and zero-mean-normalized cross correlation techniques, which extracts depth information using coarse-to-fine disparity between corresponding windows in a stereo pair. The gray level encoded sparse disparity matrix is subjected to a cubic B-spline operation to generate smooth representations of the optic cup/disc surfaces and new three-dimensional (3-D) metrics from isodisparity contours. Despite the challenges involved in 3-D surface recovery, the robustness of our algorithm in finding disparities within the constraints used has been validated using stereo pairs with known disparities. In a preliminary longitudinal study of glaucoma patients, a strong correlation is found between the computer-generated quantitative cup/disc volume metrics and manual metrics commonly used in a clinic. The computer generated new metrics, however, eliminate the subjective variability and greatly reduce the time and cost involved in manual metric generation in follow-up studies of glaucoma. PMID- 12585707 TI - A novel measurement technique to assess the effects of coronary brachytherapy in clinical trials. AB - This paper presents a novel measurement technique to assess the effects of coronary brachytherapy. This new technique is based upon the conventional quantitative coronary analysis (QCA) technique, which is accepted worldwide as an accurate and reliable analysis tool for clinical trials. This paper provides the definitions and main issues important for correct brachytherapy analysis. Based on these definitions, this novel technique is implemented as an extension of conventional QCA software, as a multisegmental analysis tool. It allows to follow the influence of radiation on restenosis, and the mutual relation between intervention devices. A pilot interobserver study was performed to assess the reliability and reproducibility of the brachytherapy analysis tool, using 15 patient cases. The validation results show that the segment lengths, minimum lumen diameter, and reference diameters of the user-defined and derived (sub)segments can be assessed reproducible. However, these good results can only be obtained, when strict and extensive image acquisition and image analysis protocols are followed. From this pilot validation study presented in this paper and only based on a small number of patients, we may conclude that the software can be applied to clinical trials. PMID- 12585708 TI - Method for three-dimensional data registration from disparate imaging modalities in the NOGA Myocardial Viability Trial. AB - Region-by-region comparison of data concerning left ventricular (LV) status is difficult to perform quantitatively if the data was acquired from disparate imaging modalities. We validated a method for comparing measurements obtained by electromechanical mapping (EMM) catheter with dobutamine stress echocardiography (DSE) via biplane contrast ventriculography, with the assistance of three dimensional (3-D) echocardiographic data. The ventriculograms were traced and the borders were used to reconstruct the LV in 3-D with the aid of a database of 3-D echocardiographic studies. The 3-D LV was oriented to the EMM data based on the body coordinates and then manually scaled and translated to fit. The EMM data were mapped to the 3-D surface. The 3-D surface was divided into the 16 regions defined for echocardiographic assessment. The mean EMM value for local linear shortening, a parameter of function, was computed in each segment. The EMM and semiquantitative echocardiographic assessments of regional myocardial function were compared by segment, and the volume of the 3-D LV was compared with the volume computed from the ventriculogram. The volume of the 3-D surface correlated closely with that of the ventriculogram (r = 0.97, SEE = 27.4 ml) but with a significant overestimation of 63 +/- 35 ml. There was a highly significant (p < 0.0001) agreement in regional function between EMM and echo. Local linear shortening correlated significantly (p < 0.0001) with echocardiographic severity of wall motion, averaging 9.5 +/- 6.5, 8.1 +/- 5.4, 5.9 +/- 4.8, and 6.2 +/- 3.3 in segments read as normal, hypokinetic, akinetic, and dyskinetic, respectively. The method presented is valid for comparing cardiac parameters derived from disparate image data on a region-by-region basis by employing anatomic landmarks on 3-D reconstructions of the LV endocardial surface. PMID- 12585709 TI - Automated analysis of brachial ultrasound image sequences: early detection of cardiovascular disease via surrogates of endothelial function. AB - Early detection of cardiovascular disease would allow timely institution of preventive measures. Arterial endothelium play a primary role in processes leading to the development of atherosclerotic plaque and cardiovascular disease in general. Determination of flow-mediated dilatation (FMD) of brachial arteries from B-mode ultrasound image sequences offers a noninvasive surrogate index of endothelial function. A highly automated method for analysis of brachial ultrasound image sequences is reported and its performance assessed. The method overcomes the variability of brachial ultrasound images across subjects by incorporating machine learning and quality control steps. The automated method outperformed conventional manual analysis by providing a decreased analysis bias, increased reproducibility, and improved measurement accuracy. Consequently, it decreases inter- and intraobserver as well interinstitution variability. The method has been employed in a number of population studies with thousands of subjects analyzed. PMID- 12585710 TI - Automatic "pipeline" analysis of 3-D MRI data for clinical trials: application to multiple sclerosis. AB - The quantitative analysis of magnetic resonance imaging (MRI) data has become increasingly important in both research and clinical studies aiming at human brain development, function, and pathology. Inevitably, the role of quantitative image analysis in the evaluation of drug therapy will increase, driven in part by requirements imposed by regulatory agencies. However, the prohibitive length of time involved and the significant intraand inter-rater variability of the measurements obtained from manual analysis of large MRI databases represent major obstacles to the wider application of quantitative MRI analysis. We have developed a fully automatic "pipeline" image analysis framework and have successfully applied it to a number of large-scale, multicenter studies (more than 1,000 MRI scans). This pipeline system is based on robust image processing algorithms, executed in a parallel, distributed fashion. This paper describes the application of this system to the automatic quantification of multiple sclerosis lesion load in MRI, in the context of a phase III clinical trial. The pipeline results were evaluated through an extensive validation study, revealing that the obtained lesion measurements are statistically indistinguishable from those obtained by trained human observers. Given that intra- and inter-rater measurement variability is eliminated by automatic analysis, this system enhances the ability to detect small treatment effects not readily detectable through conventional analysis techniques. While useful for clinical trial analysis in multiple sclerosis, this system holds widespread potential for applications in other neurological disorders, as well as for the study of neurobiology in general. PMID- 12585711 TI - Quantification of small cerebral ventricular volume changes in treated growth hormone patients using nonrigid registration. AB - Nonrigid registration can automatically quantify small changes in volume of anatomical structures over time by means of segmentation propagation. Here, we use a nonrigid registration algorithm based on optimising normalized mutual information to quantify small changes in brain ventricle volume in magnetic resonance (MR) images of a group of five patients treated with growth hormone replacement therapy and a control group of six volunteers. The lateral ventricles are segmented from each subject image by registering with the brainweb image which has this structure delineated. The mean (standard deviation) volume change measurements are 1.09 (0.73) cm3 for the patient group and 0.08 (0.62) cm3 for the volunteer group; this difference is statistically significant at the 1% level. We validate our volume measurements by determining the precision from three consecutive scans of five volunteers and also comparing the measurements to previously published volume change estimates obtained by visual inspection of difference images. Results demonstrate a precision of sigma < or = 0.52 cm3 (n = 5) and a rank correlation coefficient with assessed difference images of p = 0.7 (n = 11). To determine the level of shape correspondence we manually segmented subject's ventricles and compared them to the propagations using a voxel overlap similarity index, this gave a mean similarity index of 0.81 (n = 7). PMID- 12585712 TI - A new deformable model for analysis of X-ray CT images in preclinical studies of mice for polycystic kidney disease. AB - This paper describes the application of a new probabilistic shape and appearance model (PSAM) algorithm to the task of detecting polycystic kidney disease (PKD) in X-ray computed tomography images of laboratory mice. The genetically engineered PKD mouse is a valuable animal model that can be used to develop new treatments for kidney-related problems in humans. PSAM is a statistical-based deformable model that improves upon existing point distribution models for boundary-based object segmentation. This new deformable model algorithm finds the optimal boundary position using an objective function that has several unique characteristics. Most importantly, the objective function includes both global shape and local gray-level characteristics, so optimization occurs with respect to both pieces of information simultaneously. PSAM is employed to segment the mouse kidneys and then texture measurements are applied within kidney boundaries to detect PKD. The challenges associated with the segmentation non-rigid organs along with the availability of a priori information led to the choice of a trainable, deformable model for this application. In 103 kidney images that were analyzed as part of a preclinical animal study, the mouse kidneys and spine were segmented with an average error of 2.4 pixels per boundary point. In all 103 cases, the kidneys were successfully segmented at a level where PKD could be detected using mean-of-local-variance texture measurements within the located boundary. PMID- 12585714 TI - Immunofluorescence imaging as a tool for studying the pharmacokinetics of a human monoclonal single chain fragment antibody. AB - We have used immunofluorescence imaging to study binding of a monoclonal antibody fragment to subcutaneously implanted human melanoma cells in nude mice. The data acquired using this nontraditional approach was then analyzed using standard pharmacokinetic methods to produce estimates of k(e) (0.06h(-1)), t 1/2 (16 h), mean residency time (23.4 h) and percent exposure of the antibody to the tumor (40%). To our knowledge this is the first time standard pharmacokinetic analyzes have been conducted on immunofluorescence imaging data. The combination of this novel imaging technique and standard pharmacokinetic analytical methods should prove to be a useful tool for comparing the properties of antibody fragments in animal models. PMID- 12585713 TI - X-ray computed tomography methods for in vivo evaluation of local drug release systems. AB - Recent advances in drug delivery techniques have necessitated the development of tools for in vivo monitoring of drug distributions. Gamma emission imaging and magnetic resonance imaging suffer from problems of resolution and sensitivity, respectively. We propose that the combination of X-ray CT imaging and image analysis techniques provides an excellent method for the evaluation of the transport of platinum-containing drugs from a localized, controlled release source. We correlated local carboplatin concentration with CT intensity, producing a linear relationship with a sensitivity of 62.6 microg/mL per Hounsfield unit. As an example application, we evaluated the differences in drug transport properties between normal and ablated rabbit liver from implanted polymer millirods. The use of three-dimensional visualization provided a method of evaluating the placement of the drug delivery device in relation to the surrounding anatomy, and registration and reformatting allowed the accurate comparison of the sequence of temporal CT volumes acquired over a period of 24 h. Taking averages over radial lines extending away from the center of the implanted millirods and integrating over clinically appropriate regions, yielded information about drug release from the millirod and transport in biological tissues. Comparing implants in normal and ablated tissues, we found that ablation prior to millirod implantation greatly decreased the loss of drug from the immediate area, resulting in a higher average dose to the surrounding tissue. This work shows that X-ray CT imaging is a useful technique for the in vivo evaluation of the pharmacokinetics of platinated agents. PMID- 12585715 TI - PET imaging-based evaluation of aerosol drugs and their delivery devices: nasal and pulmonary studies. AB - Three-dimensional (3-D) positron emission tomography (PET) imaging of inhaled radiolabeled aerosol in the nasal or pulmonary regions provides an in vivo measurement of drug distribution using the drug itself as the tracer. Repeated or dynamic PET scans over the time after inhalation provides us with further information about the fate of the deposited drug. These quantitative measurements are sufficient to describe the performance of a drug or device and they are obtained in a noninvasive fashion, which cannot be achieved by using any other methods. Using this PET-imaging paradigm, we conducted a sequence of drug studies to evaluate the performance of aerosol drugs and delivery devices; to compare the performance of similar drugs from different manufacturers; to assess the similarity between different formulations and propellants for the same drug; to appraise delivery devices such as spacers and nebulizers, etc. This paper reviews only the imaging and data analysis techniques developed for the above-mentioned studies that include multi-modality image registration, region definition and region-based data analysis, and nonregion-based data analysis. We separated the techniques into nasal and pulmonary studies because of the uniqueness of each group. Specific drugs or devices are not identified and no result about drug performance is given because the imaging and data analysis methodology, which is the focus of this paper, applies to all these studies regardless of the drugs or their delivery devices. The quantitative data are used as the scientific basis for evaluation although we also developed visualization techniques to enhance the results drawn from the data. PMID- 12585716 TI - Oxidative stress in the brain of Fukuyama type congenital muscular dystrophy: immunohistochemical study on astrocytes. AB - Astrocytes in the cerebrum and medulla oblongata of cases of Fukuyama type congenital muscular dystrophy were examined by immunohistochemistry of oxidative modification products and free-radical scavenging enzymes because abnormal glia limitans formed by astrocytic end feet is considered to be involved in the genesis of brain lesions of Fukuywama type congenital muscular dystrophy. The study was performed on two fetal cases of Fukuyama type congenital muscular dystrophy of 18 and 20 weeks' gestation and seven patients with Fukuyama type congenital muscular dystrophy ranging in age from 2 to 27 years. Eight age matched control cases were used. Polymerase chain reaction (PCR) was performed to ascertain the gene phenotype of two child cases, in which prenatal gene analysis was not performed. Astrocytes, especially layer I astrocytes, of postnatal cases of Fukuyama type congenital muscular dystrophy were weakly positivefor Nepsilon (carboxymethyl)lysine and argpyrimidine, suggesting that they were sensitive to oxidative stress, and the accumulation may be related to the abnormal glia limitans. Secondary increase of manganese (Mn) superoxide dismutase against the increase of free radicals was considered in patients with Fukuyama type congenital muscular dystrophy more than 14 years old considered to be homozygous for founder haplotype: homozygosity was suggested by PCR in two cases. In contrast, expression of Mn superoxide dismutase was decreased in 2- and 6-year old children with Fukuyama type congenital muscular dystrophy that were heterozygous. Moreover, accumulation of argpyrimidine was exclusively found in astrocytes of the 2-year-old child that exhibited severe brain lesions. Function of astrocytes might be impaired or immature in severe or heterozygous cases. These results may confirm that astrocytes play an important role in the etiology of the brain lesion. PMID- 12585717 TI - Epilepsy, occipital calcifications, and oligosymptomatic celiac disease in childhood. AB - The association of epilepsy, occipital calcifications, and celiac disease has been recognized as a distinct syndrome. The objective of this study was to present the clinical, electrophysiologic, and neuroradiologic features in a series of patients with this syndrome. Thirty-two patients with the constellation of epilepsy, occipital calcifications, and celiac disease were identified in our epilepsy clinic. The mean age was 11 years and the mean length of follow-up was 7.4 years. The 1990 criteria of the European Society of Pediatric Gastroenterology and Nutrition were used to diagnose celiac disease. The Kruskal Wallis statistics test was employed with a signficance of P < .05. Thirty-one patients had partial seizures, 21 of them with symptoms related to the occipital lobe. In most patients, the epilepsy was controlled or the seizures were sporadic. Three developed severe epilepsy. Occipital calcifications were present in all cases. Computed tomography in 7 patients showed hypodense areas in the white matter around calcifications, which decreased or disappeared after a period of gluten-free diet in 3 patients. A favorable outcome of epilepsy was detected in patients with the earliest dietary therapy. This study presents the largest series of children with this syndrome outside Italy. White-matter hypodensities surrounding calcifications are rarely reported. A prompt diagnosis of celiac disease might improve the evolution of the epilepsy and may improve cognitive status. PMID- 12585718 TI - Value of neuroimaging in the evaluation of neurologically normal children with recurrent headache. AB - Headache is one of the most frequent physical complaints in children. Although headaches in children are generally benign, neuroimaging studies are frequently performed in clinical practice for the fear of missing a serious underlying disease. Despite this, limited data exist about the utility of neuroimaging in recurrent headache of children with a normal neurologic examination. This prospective study was planned to determine the value of neuroimaging in neurologically normal children with migraine and tension-type headache. Among 95 consecutive patients presenting with headache, 72 patients receiving a diagnosis of migraine or tension-type headache were included in the study. Neuroimaging procedures were performed in 83%. Magnetic resonance imaging (MRI) was abnormal in 11 of 49 cases. Abnormalities consisted of foci of gliosis in four, sinusitis in two, pineal cyst in one, periventricular leukomalacia in one, arachnoid cyst in one, old traumatic changes in one, and cervical syrinx in one. Two of the 11 computed tomographic (CT) scans revealed sinus disease. The percentage of findings causally related to headache was about 10. None of the patients had undergone surgery because of neuroimaging results. In conclusion, the yield of neuroimaging in recurrent headaches of children with a normal neurologic examination is low, and neuroimaging should not be part of a routine initial examination of these patients. PMID- 12585719 TI - Outcome of severe encephalomyelitis in children: effect of high-dose methylprednisolone and immunoglobulins. AB - Acute encephalomyelitis in children refers to an insult of cortical white matter leading to acute disseminated encephalomyelitis, insult of the spinal cord leading to multifocal myelopathy, or a combined form of encephalomyelitis. We report here the clinical presentations and outcome of 16 children with severe acute encephalomyelitis analyzing the effect of high-dose methylprednisolone or intravenous immunoglobulins, administered separately or in combination. Five children developed acute disseminated encephalomyelitis alone, eight developed severe multifocal myelopathy accompanied in two of them by radiculoneuropathy, and three developed the most severe form of combined encephalomyeloradiculoneuropathy. The indications for treatment with either high dose methylprednisolone, intravenous immunoglobulin, or a combination of the two were severe acute disseminated encephalomyelitis, visual loss, or severe flaccid weakness accompanied by bladder and bowel incontinence. Overall, 10 children had remarkably responded to high-dose methylprednisolone alone and recovered within 10 days. One patient with severe myelopath, developing paraplegia, who failed oral corticosteroids completely recovered following intravenous immunoglobulin. Of the isolated acute disseminated encephalomyelitis group, all patients were initially treated with high-dose intravenous methylprednisolone and recovered within 10 days, including visual remission in the child with severe optic neuritis. All six children with solitary severe multifocal myelopathy were treated with high-dose methylprednisolone alone and recovered within the first week. Two patients had severe myeloradiculoneuropathy and were therefore treated with combined high-dose methylprednisolone and intravenous immunoglobulin: one remains paraplegic, whereas the second was ventilated for 3 weeks and recovered after 2 months. The three children with the most severe form of encephalomyeloradiculoneuropathy were treated with combined high-dose methylprednisolone and intravenous immununoglobulin; two remain severely handicapped, of whom one is paraplegic, and the third unexpectedly recovered within 3 months. Therefore, our experience indicates that either high-dose methylprednisolone or intravenous immunoglobulin, given separately or combined, may be efficacious in severe debilitating pediatric-onset acute encephalomyelitis. In children with the most severe form of encephalomyeloradiculoneuropathy, we suggest initially administering high-dose methylprednisolone and intravenous immunoglobulin combined, given the poorer outcome of our patients with combined severe central and peripheral demyelination. PMID- 12585720 TI - Cerebrospinal fluid levels of nitric oxide and nitrotyrosine in neonates with mild hypoxic-ischemic encephalopathy. AB - The objective of this study was to determine the role of cerebral nitric oxide and its powerful oxidant peroxynitrite following mild birth asphyxia. The cerebrospinal fluid levels of nitric oxide and 3-nitrotyrosine as a marker for peroxynitrite are measured in neonates with mild hypoxic-ischemic encephalopathy. Based on the classification of Sarnat and Sarnat, term neonates with mild hypoxic ischemic encephalopathy and neurologically normal neonates suspected of sepsis were taken as the control group. Nitric oxide measurements were done by chemiluminescence, and nitrotyrosine measurements were made by high-performance liquid chromatography. The Mann Whitney U-test was used, and a Pvalue < .05 was considered significant. Eleven patients with grade 1 hypoxic-ischemic encephalopathy and nine controls were included. The gestational age and birthweights were similar in both groups. Neither of the cerebrospinal fluid levels of nitric oxide (8.60 +/- 0.49 micromol/L) and nitrotyrosine (0.45 +/- 0.33 micromol/L) of the neonates with hypoxic-ischemic encephalopathy showed significant differences from that of the means of nitric oxide (8.66 +/- 1.07 micromol/L) and nitrotyrosine levels (0.25 +/- 0.13 micromol/L) of the controls. These data suggest that the oxidative stress is not overexpressed to lead nitric oxide and peroxynitrite to play a pathologic role in the early phase of mild hypoxic-ischemic encephalopathy of the newborn. PMID- 12585722 TI - Neuron-specific enolase levels and neuroimaging in asphyxiated term newborns. AB - The study was designed to investigate the cerebrospinal fluid and serum levels of neuron-specific enolase, along with cranial ultrasonography, magnetic resonance imaging (MRI), and electroencephalography (EEG), for predicting the clinical state and neurologic outcome of 26 asphyxiated term newborns. The babies were graded according to the Sarnat and Sarnat classification. Cerebrospinal fluid neuron-specific enolase levels of the 18 babies in the whole hypoxic-ischemic encephalopathy group were higher than the 8 babies in the "no encephalopathy" group. Cerebrospinal fluid neuron-specific enolase levels of 13 cases in the hypoxic-ischemic encephalopathy grade 2 and 3 groups (high-risk group) were higher than both the no encephalopathy and hypoxic-ischemic encephalopathy grade 1 groups when pooled. Cerebrospinal fluid neuron-specific enolase levels of the 7 newborns in the hypoxic-ischemic encephalopathy grade 3 group were also significantly higher than the 5 in the hypoxic-ischemic encephalopathy grade 1 group. The findings of cranial MRI, EEG, and cerebrospinal fluid neuron-specific enolase levels were correlated with each other and the clinical grade of the patients and also were predictive of the neurologic outcome at 1 year of age. Cerebrospinal fluid neuron-specific enolase levels, cranial MRI, and EEG are predictive of outcome of hypoxic-ischemic brain damage in asphyxiated newborns, and this predictivity would increase with the combination of these diagnostic parameters. PMID- 12585721 TI - Childhood-onset chronic inflammatory demyelinating polyradiculoneuropathy with cranial nerve involvement. AB - A 17-year-old male presented with chronic diplopia and generalized motor weakness. He was previously diagnosed with chronic inflammatory demyelinating polyradiculoneuropathy and acute disseminated encephalomyelitis in childhood. Cranial magnetic resonance imaging (MRI) revealed a rarely reported finding of thickening and enhancement of multiple cranial nerves. Nerve conduction studies and electromyography showed peripheral nerve demyelination with axonal involvement. There was improvement in the clinical examination, MRI, and electrophysiologic studies after combined corticosteroid and plasma exchange therapy. We review the clinical presentation, course, and response to therapy among children with chronic inflammatory demyelinating polyradiculoneuropathy, with specific emphasis on the frequency and pattern of cranial nerve involvement. PMID- 12585723 TI - Supratentorial tuber location and autism in tuberous sclerosis complex. AB - The high rate of autism in tuberous sclerosis complex provides an opportunity to study the pathogenesis of autism. This study investigated the relationship between a DSM-IV diagnosis of autism and tuber location in a sample of 50 individuals with tuberous sclerosis complex. Chi-square analyses revealed no differences between individuals with autism (n = 15) and those without autism (n = 35) on the occurrence of tubers in the right or left frontal, occipital, parietal, or temporal regions. There were no differences between the two groups in the occurrence of tubers in subcortical or cortical regions. In the largest sample to date, these results fail to support the hypothesis that supratentorial tuber location is a marker for autism. PMID- 12585724 TI - Double-blind, placebo-controlled study of L-carnosine supplementation in children with autistic spectrum disorders. AB - L-Carnosine, a dipeptide, can enhance frontal lobe function or be neuroprotective. It can also correlate with gamma-aminobutyric acid (GABA) homocarnosine interaction, with possible anticonvulsive effects. We investigated 31 children with autistic spectrum disorders in an 8-week, double-blinded study to determine if 800 mg L-carnosine daily would result in observable changes versus placebo. Outcome measures were the Childhood Autism Rating Scale, the Gilliam Autism Rating Scale, the Expressive and Receptive One-Word Picture Vocabulary tests, and Clinical Global Impressions of Change. Children on placebo did not show statistically significant changes. After 8 weeks on L-carnosine, children showed statistically significant improvements on the Gilliam Autism Rating Scale (total score and the Behavior, Socialization, and Communication subscales) and the Receptive One-Word Picture Vocabulary test (all P < .05). Improved trends were noted on other outcome measures. Although the mechanism of action of L-carnosine is not well understood, it may enhance neurologic function, perhaps in the enterorhinal or temporal cortex. PMID- 12585725 TI - Coincidence of rolandic and absence features: rare, but not impossible. AB - The purpose of the study was to evaluate the incidence of a possible combination of rolandic and absence signs in epileptic children, which remains a subject of controversy. The medical files and electroencephalographic (EEG) records of children with rolandic, childhood absence, and juvenile absence epilepsy were retrospectively analyzed. During the antiepilepsy treatment, 6 of 66 patients with rolandic epilepsy, most of them with initial carbamazepine therapy, had absences and generalized spike-wave discharges of a secondarily generalized type. Five cases of 34 children with childhood absence epilepsy and 3 of 11 patients with juvenile absence epilepsy were identified with an EEG focus of the rolandic type. We considered the likely relation of absence features in rolandic epilepsy to the treatment or to an atypical course. The presence of a rolandic focus in absence epilepsies, however, makes the coincidence of these entirely distinct phenomena, even if very rare, not excluded. Further studies are required to elucidate a probable genetic or functional link between partial and primarily generalized EEG discharges in the idiopathic childhood epilepsies. PMID- 12585726 TI - Cobb syndrome: case report. AB - We present a 7-year-old boy in whom cutaneous hemangioma and intramedullary and paraspinal arteriovenous malformations were associated. Magnetic resonance arteriography revealed the presence of the two arterovenous malformations, and the selective intercostal arteriography demonstrated that the intraspinal and paraspinal arteriovenous malformations were supplied by the same intercostal arteries. Rubbing the back was required to detect the cutaneous changes, which were only suspected by casual inspection. PMID- 12585727 TI - Isolated abducens nerve palsy: an unusual presentation of leukemia. AB - We present a case of isolated abducens nerve palsy with negative magnetic resonance imaging (MRI) findings and unusual cerebrospinal fluid results. The patient was treated for a central nervous system infection initially. After the second cerebrospinal fluid examination, a diagnosis of central nervous system leukemia was made. Isolated abducens nerve palsy is not always a stable or benign illness, even with a normal brain MRI study. Abnormal cerebrospinal fluid results should suggest the presence of a central nervous system malignancy. PMID- 12585728 TI - Magnetoencephalographic analysis in a case of early-onset benign childhood occipital seizures. AB - We report a 12-year-old boy who had one seizure comprising deviation of the eyes followed by impairment of consciousness for 30 minutes at the age of 6 years. No visual hallucination or ictal vomiting was observed. Interictal electroencephalography showed repetitive spikes and spike-wave discharges over the left occipital lobe. Magnetoencephalography revealed that the estimated dipoles were clustered in the left cuneus on magnetic resonance imaging, which corresponded to the area of peripheral vision and the associated visual cortex but not to the central visual cortex. Magnetoencephalography is advantageous for determining the electrophysiologic mechanism of early-onset benign childhood occipital seizures. PMID- 12585729 TI - Calcium carbonate consumption during pregnancy: an unusual cause of neonatal hypocalcemia. AB - The occurrence of hypocalcemia in a term infant after the first few days of life is known to result from excessive dietary phosphate, hypomagnesemia, neonatal hypoparathyroidism, hypovitaminosis D, and maternal hyperparathyroidism. Less well recognized is the possibility that maternal ingestion of antacids may cause late neonatal hypocalcemia. Our observations suggest that taking large doses of calcium carbonate during pregnancy may be a potential cause of neonatal hypocalcemia. PMID- 12585730 TI - Pica and the elephant's ear. AB - This is a case report of an otherwise healthy 2-year-old boy with a history of pica, associated with iron deficiency anemia. This boy was referred to our department for a neurologic evaluation because of an acute episode of sialorrhea, difficulty in speaking, dysphagia, and repeated swallowing movements. An uncertain episode of a brief-duration still gaze was also reported. In addition, the history revealed that the child had earlier ingested a leaf from a poisonous houseplant called Colocasia esculenta, also known as "elephant's ear." The habit of pica subsided after treatment with iron supplements. A 9-month follow-up period was uneventful. Neurologic manifestations can accompany accidental intoxications of some non-nutrient substances. Thus, pica must be suspected in children with acute behavior alterations. PMID- 12585731 TI - Migraine-type headaches in children receiving chemotherapy and ondansetron. AB - Six children developed severe daily migraine-type headaches during cancer treatment. In addition to chemotherapy drugs, all received daily doses of ondansetron, a 5-hydroxytryptamine type 3 receptor antagonist. 5 Hydroxytryptamine is considered to play a central role in migraine pathogenesis, and ondansetron may have caused headaches by producing 5-hydroxytryptamine dysfunction in the brain. All six children had either a personal or a family history of migraine, and this may be a risk factor for developing ondansetron associated migraine-type headaches. Ondansetron-induced headaches respond to withholding the drug and to standard antimigraine medications, but further study of a larger group of patients is required to confirm this impression. PMID- 12585734 TI - Clinical practice guidelines--past, present, and future. PMID- 12585733 TI - The cost of doing business. PMID- 12585732 TI - Homocystinuria presenting as psychosis in an adolescent. AB - Homocystinuria usually presents with ectopia lentis, mental retardation, thromboembolic complications, and skeletal abnormalities. Whereas neuropsychiatric abnormalities are often recognized in untreated homocystinuria, initial presentation with acute psychosis has only rarely been reported. We describe a previously well 17-year-old adolescent with an acute psychosis characterized by auditory and visual hallucinations and marked paranoia who was found to have pyridoxine-responsive homocystinuria. His mental state normalized within several weeks of inception of pyridoxine and antipsychotic therapy. Pyridoxine-responsive homocystinuria is commonly missed on neonatal screens and should be recognized as a potentially treatable cause of acute psychosis in childhood and adolescence. PMID- 12585735 TI - Alzheimer's: diagnosis and intervention. PMID- 12585736 TI - Chemical, biological, and radiological weapons: implications for optometry and public health. AB - BACKGROUND: Because of growing concerns of terrorist attacks using chemical, biological, radiological, and nuclear weapons, appropriate diagnosis and management of ocular and systemic effects are important issues for eye care practitioners. OVERVIEW: The literature on the ocular and systemic effects of chemical, biological, radiological, and nuclear weapons is reviewed to provide practicing optometrists with an overview of the issues involved in incident management. A discussion of the public health issues and implications is also presented. Chemical, biological, radiological, and nuclear weapons can exert serious consequences on the eye and the entire body. Acts of terrorism can pose unique challenges to patient care and to the healthcare system infrastructure. The clinician must be familiar with signs of potential terrorist activities to aid in appropriate patient care and in the containment of illness in the community. CONCLUSIONS: Knowledge of the signs and symptoms of exposure to potential agents used by terrorists and a review of the public health concerns of managing terrorist events can provide the necessary background to help the eye care provider make the appropriate diagnosis and understand the role of the clinician in the hierarchy of disaster preparedness, surveillance, and response. PMID- 12585737 TI - Cigarette smoking as a risk factor for cataract and age-related macular degeneration: a review of the literature. AB - BACKGROUND: Cigarette smoking has long been acknowledged as a major risk factor for many of the leading causes of death. Its role in ocular disease has received far less attention. METHODS: This article reviews a large number of recent studies that have investigated a possible link between cigarette smoking and two major causes of blindness--cataract and age-related macular degeneration. RESULTS: The preponderance of evidence shows a causal relationship between cigarette smoking and both cataract and age-related macular degeneration. CONCLUSIONS: As primary eye care providers, optometrists should educate patients regarding the dangers of smoking as it relates to ocular health and consider a role in discouraging patients from starting or continuing this harmful habit. PMID- 12585738 TI - Post-traumatic pseudomyopia. AB - BACKGROUND: Many clinicians have noted that patients demonstrate a myopic refractive change following Traumatic Brain Injury (TBI). This apparent myopic shift disappears with cycloplegia, yet stubbornly reappears as soon as the pharmaceutical effect wears off. We propose that this shift is secondary to an irritative lesion that affects the parasympathetic innervation, resulting in ciliary body contracture. The dilemma for the clinician is whether to provide the immediate relief of clear distance vision by prescribing additional minus lenses, or to work toward attempting to re-establish the baseline refractive error. CASE REPORTS: The natural history of post-traumatic pseudomyopia in our experience involves one of the following three courses: (1) a transient condition that will occasionally resolve; (2) the typical case, a recalcitrant condition that will resolve under cycloplegic intervention, but immediately return as the cycloplegic wears off; or (3) a less-common subgroup of patients who continue to show an increase in myopia over time. Our description of these cases demonstrates management strategies (including atropinization) to relax accommodative spasm, traditional vision therapy techniques aimed at loosening the accommodative system, and refractive corrections. CONCLUSIONS: Pseudomyopia is one of many ocular and behavioral sequelae following TBI. By understanding the natural course and potential management options for post-traumatic pseudomyopia, the clinician will be better prepared to deal with these challenging cases. Flexibility is required, since options that work with one patient may prove ineffective with another. Counseling the patient as to potential outcomes given the natural history of this condition helps establish more-realistic expectations by the patients being treated. PMID- 12585739 TI - Don't put off employee performance appraisals. PMID- 12585740 TI - The antitrust laws and optomety practice. PMID- 12585741 TI - Healthy eyes healthy people: the practitioner's role. PMID- 12585742 TI - Guidelines for laboratory testing and result reporting of antibody to hepatitis C virus. Centers for Disease Control and Prevention. AB - Testing for the presence of antibody to hepatitis C virus (anti-HCV) is recommended for initially identifying persons with hepatitis C virus (HCV) infection (CDC. Recommendations for prevention and control of hepatitis C virus [HCV] infection and HCV-related chronic disease. MMWR 1998;47[No. RR-19] :1-33). Testing for anti-HCV should include use of an antibody screening assay, and for screening test-positive results, a more specific supplemental assay. Verifying the presence of anti-HCV minimizes unnecessary medical visits and psychological harm for persons who test falsely positive by screening assays and ensures that counseling, medical referral, and evaluation are targeted for patients serologically confirmed as having been infected with HCV. However, substantial variation in reflex supplemental testing practices exists among laboratories, and an anti-HCV-positive laboratory report does not uniformly represent a confirmed positive result. These guidelines expand recommendations for anti-HCV testing to include an option for reflex supplemental testing based on screening-test positive signal-to-cut-off (s/co) ratios. Use of s/co ratios minimizes the amount of supplemental testing that needs to be performed while improving the reliability of reported test results. These guidelines were developed on the basis of available knowledge of CDC staff in consultation with representatives from the Food and Drug Administration and public health, hospital, and independent laboratories. Adoption of these guidelines by all public and private laboratories that perform in vitro diagnostic anti-HCV testing will improve the accuracy and utility of reported anti-HCV test results for counseling and medical evaluation of patients by health-care professionals and for surveillance by public health departments. PMID- 12585744 TI - A new era in pharmacy school education. PMID- 12585743 TI - Where is technology taking pharmacy? (Better yet, where is pharmacy taking technology?). PMID- 12585745 TI - The latest impact of technology on pharmacy students. PMID- 12585746 TI - Filling a prescription for the public's health. PMID- 12585747 TI - Autopsy: coaxing secrets from the dead. Every drug is a poison; every poison, a drug. PMID- 12585748 TI - Extent of services provided by pharmacists in the Iowa Medicaid Pharmaceutical Case Management program. AB - OBJECTIVE: To summarize the start-up experience with patients identified as eligible during the first four quarters of the Iowa Pharmaceutical Case Management (PCM) program and to characterize the extent of the services proved by pharmacists in the program. DESIGN: Prospective pharmaceutical care intervention project. SETTING: One hundred seventeen community pharmacies in Iowa. PATIENTS: Medicaid patients at high risk for drug-related problems based on using 4 or more nontopical medications and having 1 of 12 specific disease states. INTERVENTION: To become eligible to provide PCM services, licensed pharmacists had to undergo training and submit five care plans to the Iowa Department of Human Services. Community pharmacists were provided names of newly eligible patents each calendar quarter for 1 year. For each patient, pharmacists were asked to indicate by fax whether they had met with the patient, performed a written work-up of the patient, sent recommendations to the patient's physician, and whether the physician replied. When pharmacists were unable to provide the service, they were asked to state the reason. Both the pharmacist and the physician receive $75 for the initial assessment, with additional payments after each follow-up visit performed. MAIN OUTCOME MEASURES: An intensity score and the percentage of eligible patients for whom all steps were completed were calculated for each pharmacy. RESULTS: Fax survey results were retumed for 2,834 (96.7%) of the 2,931 patients eligible for PCM services. Pharmacists met with 943 (33.3%), worked up 763 (26.9%), sent recommendations to physicians for 500 (17.6%), and received replies from physicians for 327 (11.5%) patients. Pharmacists were unable to provide PCM services for 1,891 (66.7%) patients. The primary reasons given for this inability to provide services were patient access issues for 438 (23.2%) patients, pharmacy staffing or start-up issues for 419 (22.2%) patients, or no reason specified for 575 (30.4%) patients. A PCM intensity score was developed to represent the scope of services provided and the number of patients served. A higher intensity score indicated pharmacies that provided PCM to more patients and/or that offered higher levels of care (e.g., provided a written set of recommendations to the physician rather than simply assessing the patient without preparing or sending recommendations). Future evaluations will determine the validity of the score on the basis of patient outcomes. CONCLUSION: Some pharmacies implemented PCM services very effectively. However, 40% to 60% of the pharmacies provided little or no PCM services within 3 months of notification of patient eligibility. Future investigations will evaluate the quality of prescribing and quality of life for patients who received PCM services. PMID- 12585749 TI - Pain management content in curricula of U.S. schools of pharmacy. AB - OBJECTIVES: To identify individuals in schools of pharmacy in the United States who are responsible for covering the topic of pain management in courses for doctor of pharmacy students and to describe how and at what depth pain management is covered in pharmacy school curricula. DESIGN: One-time qualitative assessment. SETTING: Schools of pharmacy in the United States. PARTICIPANTS Twenty-eight faculty members with the rank of professor, associate professor, or assistant professor who had been employed in their current positions for at least 2 years and who were directly involved in preparing and teaching didactic courses that address pain management. INTERVENTION: In-depth telephone interviews. MAIN OUTCOME MEASURES: Qualitative responses to open-ended interview questions. RESULTS: While pain management was included in the curricula of all 28 schools of pharmacy, it was generally covered in a fragmented way, usually as part of presentations on diseases with pain as a prominent feature (e.g., cancer pain addressed during oncology lectures) or as part of discussions of analgesics. Only two schools offered stand-alone courses in pain management, and both of those courses were electives that were taken by an average of 15 students per year. Three-fourths of respondents believed that pain was being given too little emphasis in their schools' curricula. Palliative care and the use of medications in the treatment of cancer pain was not presented in a standardized manner, and respondents were unsure of how the subject was covered in pharmacy law classes. Instruction about the diagnosis of pain, patient assessment, and physical examination was reported as "minimal" by most respondents. Respondents perceived a need for a single, complete reference and teaching resource that would address the entire spectrum of pain management as it applies to pharmacy. CONCLUSION: The topic of pain management is poorly presented and inadequately developed in the curricula of many U.S. schools of pharmacy. PMID- 12585750 TI - Identification of essential elements in the documentation of pharmacist-provided care. AB - OBJECTIVES: [corrected] To develop guidelines for the documentation elements that need to be included in any record of pharmacist-provided care to allow the quality of the care to be assessed and to describe the use of these guidelines to improve the quality of pharmacist documentation. DESIGN: An initial list of 85 potential documentation elements, developed through a review of the literature, was validated by a group of pharmacy practitioners. Then, through three rounds of a Delphi process and a group meeting, a panel of experts reached consensus on a refined list of 27 documentation data elements. RESULTS: The documentation elements were formatted into a one-page Tool for Evaluation of Documentation (TED). The TED is a checklist for assessing the completeness of the documentation of pharmacist-provided care. CONCLUSION: The TED and the consensus-building methodology used in the development of this tool can serve as cornerstones of a quality assessment process for documentation of pharmacist-provided care, enable further assessment of the quality of care, and, ultimately, be used to measure the impact of pharmacist-provided care on patient outcomes. Our results should provide guidance both to pharmacists providing care and to organizations that assess the quality of that care. PMID- 12585751 TI - Identifying at-risk patients through community pharmacy-based hypertension and stroke prevention screening projects. AB - OBJECTIVE: To demonstrate whether a community pharmacist can be successful in identifying and referring patients with elevated blood pressure and/or increased risk of stroke. SETTING: An independent community pharmacy and well-elderly housing facility in rural Iowa. PRACTICE DESCRIPTION: The pharmacy had dedicated space for patient care activities, had a community pharmacy practice resident, and served as a clerkship site for a local school of pharmacy. One of three well elderly housing facilities in the same community was used as a screening site for the stroke prevention program. PRACTICE INNOVATION: All adults entering the pharmacy duringthe time the blood pressure project was underway were offered a free blood pressure screening. If readings were elevated, patients were referred to their primary care provider. For stroke prevention, a screening using the American Heart Association stroke risk assessment protocol was held at the pharmacy and the well-elderly housing facility. MAIN OUTCOME MEASURES: Blood pressure categories and stroke risk (normal, mild, moderate, and high) categories obtained during the screening. RESULTS: A total of 351 patients werescreened for hypertension. Of these, 216 (62%) had readings greater than 140/90 mm Hg. Of the 121 patients referred to their physician, 43 (36%) had a regimen change. A total of 50 patients were screened for stroke risk. Results of the risk assessments for patients screened were normal, 4%; mild, 26%; moderate, 32%; high, 38%. CONCLUSION: These projects demonstrated that, through ongoing screening programs, community pharmacists are in an ideal position to screen patients at risk for cardiovascular and cerebrovascular disease and refer patients to their physicians for further evaluation. PMID- 12585752 TI - Pharmacist involvement in Healthy People 2010. AB - OBJECTIVES: To review opportunities through which pharmacists can help the United States achieve its public health goals as expressed in Healthy People 2010, a document issued by the federal government that expresses the areas of focus for Americans in the first decade of the 21st century. SUMMARY: Healthy People 2010 provides general goals for 10 leading health indicators (such as tobacco use, over-weight and obesity, and immunizations), and these are then further subdivided into 28 focus areas, many of them with quantifiable goals (such as, "Reduce hospitalization rates for three ambulatory care-sensitive conditions- pediatric asthma, uncontrolled diabetes, and immunization-preventable pneumonia and influenza."). As health care professionals, pharmacists have the responsibility to help the country meet these goals. Ideas for increased pharmacist involvement are described in the article, including the conduct of screening programs and provision of specialized services that focus on such areas as hypertension, diabetes, asthma, patient education, smoking cessation, or general medication management. Pharmacists can build their efforts in these and similar areas by collaborating with physicians and other appropriate professionals, identifying target patients who have obtained services at the pharmacy, contacting patients in at-risk populations within the pharmacy's patient base and/or the community, choosing and monitoring an objective of interest, and maintaining efforts for sustained time periods. CONCLUSION: The message of Healthy People 2010 is that the health of the individual is closely linked to the health of the community and hence the health of the nation. Pharmacists, uniquely positioned as the most accessible health care providers in the community, can dedicate their considerable strengths toward using Healthy People 2010 as a tool to organize their own efforts and motivate their patients. PMID- 12585753 TI - Using health observances to promote wellness in community pharmacies. AB - OBJECTIVES: To provide pharmacists in community practice a framework for using national health observances as opportunities to promote patients' health through education and screenings, to discuss obstacles pharmacists may encounter when developing services within their pharmacies, and to outline examples of activities pharmacists can perform for specific health observances. DATA SOURCES: Articles published between January 1970 and April 2002 were identified through MEDLINE using the search terms wellness, disease prevention, health promotion, Healthy People 2010, treatment of high cholesterol, treatment of high blood pressure, and levels of participation. Additional articles were identified from Web sites and reports from the federal Office of Disease Prevention and Health Promotion (ODPHP), American Heart Association, American Diabetes Association, National Osteoporosis Foundation, National Cancer Institute, American Cancer Society, National Heart, Lung, and Blood Institute, Consumer Product Safety Commission, and Centers for Disease Control and Prevention. DATA SYNTHESIS: Healthy People 2010 is a federal program with the goal of increasing the quality and years of healthy life and eliminating health disparities among populations. ODPHP publishes a list of national health observances each year. Community pharmacists can use these month- and week-long observances as opportunities to work toward achieving Healthy People 2010 goals by advocating, facilitating, and/or providing education and screenings to their patients. This article presents advice for pharmacists who want to develop pharmacy-based health promotion activities at various levels of resources and commitment. Specific suggestions include tips on preparing for and implementing education and screening programs and overcoming potential obstacles. CONCLUSION: As the most accessible health care professionals, pharmacists are in a unique position to help the nation achieve the goals of Healthy People 2010 through their involvement in the promotion of wellness. PMID- 12585754 TI - A proposed ethical framework for prescription drug benefit allocation policy. AB - OBJECTIVES: To present an ethical framework that could aid prescription drug benefit design and to propose that such a framework could be organized around a benefit allocation hierarchy. SUMMARY: Four hierarchical levels of allocation are proposed, as follows: (1) drug categories and subcategories, (2) individual drugs within covered drug categories, (3) specific indications of covered drugs, and (4) special features of drug use, such as quantities covered for specific indications. Rationales for decision making should address the distinct ethical issues apparent at each level. CONCLUSION: Ethical issues associated with designing and managing a pharmacy benefit can be identified by using a hierarchical prescription drug allocation scheme. Articulating these issues is an important first step toward arriving at a fully developed framework that will serve to produce rationales acceptable to typical stakeholders in prescription drug benefits. PMID- 12585755 TI - Issues and barriers related to the provision of pharmaceutical care in community health centers and migrant health centers. PMID- 12585756 TI - Clock drawing test and medication complexity index as indicators of medication management capacity: a pilot study. PMID- 12585757 TI - State of CAM education in U.S. schools of pharmacy: results of a national survey. PMID- 12585758 TI - Sesquicentennial Stepping Stone Summits--Summit Two: pharmacy technicians. PMID- 12585759 TI - White paper on pharmacy technicians (2002): needed changes can no longer wait. PMID- 12585760 TI - Preventing meningococcal outbreaks: an opportunity for pharmacists. PMID- 12585761 TI - New biotechnology approvals for 2002. PMID- 12585762 TI - Evaluation of OTC product information in three drug information databases for PDAs. PMID- 12585763 TI - Aiming for, believing in, and achieving a target A1c of less than 7. PMID- 12585764 TI - Zada Mary Cooper (1875-1961). PMID- 12585766 TI - Provision of behavioral health services by primary care providers in rural areas: opportunity, responsibility, and politics. PMID- 12585767 TI - Rural bioterrorism: are we exempt? PMID- 12585768 TI - Protecting rural communities from terrorism: a statewide, community-based model. AB - Given the number of Americans who live in rural areas and the unique challenges they face in the provision of health care services, special attention to planning for and responding to terrorist acts is warranted. After September 11, 2001, Florida developed a statewide, community-based model that applies the public health principles of assessment, policy development, and assurance. This model can serve as a possible framework for other states and communities. PMID- 12585769 TI - Trends in childhood drowning on U.S. farms, 1986-1997. AB - Computerized mortality data files from the National Center for Health Statistics were analyzed to describe childhood farm drowning from 1986 through 1997. Farm drowning rates were compared to the U.S. unintentional youth drowning rates for the same period. The denominator for the calculation of rates was derived from a series of farm youth estimates published by the Bureau of Census. There were 378 childhood farm drowning cases during the study period, for an average annual rate of 2.3 deaths per 100,000 farm youth resident years. This rate is comparable to unintentional drowning rates for U.S. youth (2.2/100,000 population). Fatality rates declined 28% from 1986 through 1997 (p = .0024) for farm youth and 41% for U.S. youth (p = .0001). An average 32 farm drowning incidents occur to youth annually, making drowning a legitimate concern for farm residents and visitors. PMID- 12585770 TI - Evaluation of a childhood lead questionnaire in predicting elevated blood lead levels in a rural community. AB - The accuracy of a lead screening questionnaire in predicting elevated blood lead levels was examined in a pediatric practice in a rural part of New York state. A retrospective chart review was used to collect data on children ages 9 to 24 months who presented for well-child visits. Children with both questionnaire and lead level results available in the chart were included in the study (n = 171). The mean blood lead level among all children was 1.6 microg/dl (median = 2.0 microg/dl, range 0 to 24 microg/dl). Four children (2.3%) had elevated lead levels (greater than 10 microg/dl), with levels for two of these children being greater than 20 microg/dl. Although our lead screening questionnaire was expanded from the standard 1991 CDC questionnaire by the inclusion of six additional items, it was not especially useful in predicting elevated blood lead levels above 10 microg/dl. However, the questionnaire exhibited some utility in predicting marked elevations in blood lead levels (over 20 microg/dl). Although results in other geographic areas might differ, the lead questionnaire may have value by enhancing parents' awareness of potential lead hazards in their children's environment and may prove to be more useful in areas of high risk to lead exposure. PMID- 12585771 TI - Medicaid managed behavioral health in rural areas. AB - As of 2000, 21 states had implemented Medicaid managed behavioral health (MMBH) programs for a significant portion of their rural population. It is not clear how MMBH programs may work in rural areas since they are primarily designed to control mental health utilization. In rural areas the challenge is often to enhance service delivery, not to reduce it. MMBH programs may also affect important features of rural delivery systems, including access to care and coordination of primary care and specialty mental health providers. This article describes the implementation of MMBH programs in rural areas based on an inventory of states implementing MMBH programs in rural counties conducted between June 1999 and June 2000. The experience of MMBH programs in rural areas is also described based on case studies conducted in six states. All 21 states included the general Medicaid population (Temporary Assistance for Needy Families); 17 states included special Medicaid populations (adults with serious and persistent mental illness and children with serious emotional disturbances). Slightly less than half the states integrated (carved-in) behavioral health with physical health services in serving the general Medicaid population; only one state integrated these services for the special Medicaid population. Access to mental health care in rural areas had generally not been restricted. MMBH had little impact on the linkage between primary care and mental health. Local Managed Behavioral Health Organizations, formed by public sector entities and providers, played an increasingly important role in the evolution of MMBH. PMID- 12585772 TI - Health care needs of the growing Latino population in rural America: focus group findings in one midwestern state. AB - Latinos represent nearly 13% of the U.S. population, surpassing African-Americans as the nation's largest racial/ethnic group. Many rural midwestern communities are seeing unprecedented growth in their Latino populations, creating new challenges and pressures for health and social service providers. This study is based on four focus groups conducted in three rural communities to examine concerns with health care services and access to care. Focus group analysis found several key barriers to health care access, including cost of health care services and frustration with the complexity of the U.S. health care system, as well as language and cultural issues that adversely affect patient-provider relationships. In addition, a number of impediments related to employer-sponsored health coverage were identified, including prohibitive premium costs as well as concerns about occupational injuries and access to care during work hours. The growth of the Latino population in the rural Midwest will require changes in existing health and social service systems to serve as a bridge to new systems in this country. We recommend several policy options including premium subsidies for low-wage jobs, community-based enrollment specialists for public programs, and continued research and data collection to monitor change and progress. PMID- 12585773 TI - AHEC in West Virginia: a case study. Area health education centers. AB - This case study describes the area health education centers (AHEC) program in West Virginia, spanning 30 years from a first-generation project at Charleston in 1972 (AHEC 1) to a newly funded statewide program (AHEC 2). The outcome is an evolving university-community partnership designed to meet changing work-force and community health needs in the heart of rural Appalachia. West Virginia University's (WVU's) application of the original Carnegie Commission AHEC recommendations (1970) resulted in the Charleston AHEC, now part of the Robert C. Byrd Health Sciences Center of WVU. AHEC today trains more than 135 residents and interns, and one-third of the third-year and fourth-year WVU medical students. Charleston offers clinical and continuing education for nurses, dentists, pharmacists, and allied health professionals. A health sciences library, distance learning, and a network of primary care clinics help define Charleston's unique AHEC role. This AHEC hub continues to meet the classic Carnegie goals of recruiting and retaining health professionals, and providing access to care in the original service area and statewide. Based on the Charleston experience, four new federally funded AHECs are being developed to link rural primary care residencies with the state-funded West Virginia rural health education partnerships. These rural consortia AHECs are applying the concept of community competency, a performance-based methodology, to integrate learning while achieving the goals of Healthy People 2010. PMID- 12585774 TI - A cohort study of family practice residency graduates in a predominantly rural state: initial practice site selection and trajectories of practice movement. AB - Limited information is available on long-term patterns of practice location choice among family physicians, although these patterns will significantly affect the quantity, quality, and continuity of health care provided in rural areas. From 1992 through 2000, five biennial surveys were administered to graduates of three family practice residency programs. Graduates were asked the location of their current and previous practice site locations, specific practice information, and global satisfaction with family practice. Identifiable practice sites were assigned a U.S. Department of Agriculture rural-urban continuum code based on the county in which the practice was located. The use of rural-urban continuum codes as a measure of rurality was believed to be particularly applicable to the small rural and frontier counties typical of the Great Plains region. For purposes of this study, practice locations were considered rural if they were in rural-urban continuum codes 7, 8, or 9. Of 593 graduates, 514 (87%) returned at least one questionnaire. Overall, about one-third of graduates had chosen their first practice sites in counties with rural-urban continuum codes of 7, 8, or 9. While most graduates remained in their initial practice site, almost half moved at least once. Those who left nonmetropolitan practices tended to move to less rural locations, though a few moved against this gradient. Moves away from the initial practice site occurred after a median of 3 years, and the likelihood of a first move decreased rapidly 5 to 6 years after graduation. Much remains to be learned about decisions to enter or leave practice in rural-urban continuum code 7, 8, and 9 counties. PMID- 12585775 TI - Rural nonphysician providers' perspectives on palliative care services in northwestern Ontario, Canada. AB - Most palliative care in rural remote areas is provided by nonphysicians. This paper reports a survey of interdisciplinary rural health service providers (not including physicians) to identify the strengths and weaknesses in palliative care service delivery in a rural and remote region in northwestern Ontario, Canada. Questionnaires were sent to 156 nurses, homemakers, social workers, and pastoral care workers who care for terminally ill persons and their families, and 122 were completed and returned (response rate 78%). Consistent with practice in most rural areas, 90% of respondents were generalists. Respondents identified several problems with palliative care services, including inadequate training for caregivers, inadequate support services for family and professional caregivers, inadequate human resources, and lack of organized volunteer programs. Suggestions for improvements included better education for service providers; better availability of palliative care services; more counseling and support services for patients, family members, and professionals; and greater availability of respite beds. Overall, respondents rated clients' needs as being better met than their own. The most frequently reported problems for care providers were related to the lack of supports for care provision. PMID- 12585776 TI - Initial and repeat mammography screening: different behaviors/different predictors. AB - Despite evidence of the efficacy of mammography in early detection of breast cancer, many women choose not to have a mammogram. Some women who have an initial mammogram do not return for a second one. Many others, especially minority women, and those who live in rural areas or in a low socioeconomic class, are not being screened according to recommended guidelines. The full benefits of early detection are available only if women receive regular screening. This study of 830 randomly selected women over age 50 from rural, eastern North Carolina included women who had never had a mammogram (213), women who had had a mammogram but not in the prior 2 years (89), and women who reported having had a mammogram in the previous 2 years (528). Logistic regression was used to identify significant mammography predictor variables. The results support the robust role that provider recommendation plays in encouraging mammography behavior; they also demonstrate that past behavior is also a strong predictor of future screening participation. Only 13% of the women who had never had a mammogram reported obtaining a referral from their provider compared with 79% of the women who had a mammogram in the previous 2 years. Likewise, 75% of the women who had engaged in mammography behavior in the prior 2 years expressed a positive intention to be screened in the next year compared with only 14% of those women who had never had a mammogram. Initial and repeat mammography screenings are different behaviors and are influenced by different factors. For provider counseling to be effective, the strategies employed need to consider past behavior as well as current recommendations, and they must incorporate a discussion of beliefs as well as reinforcement of regular screening behavior. PMID- 12585777 TI - Realized access to physician services among the elderly in West Texas. AB - The purpose of this study is to identify populations in a sparsely populated region that are less likely to obtain medical care. We conducted a cross sectional survey of more than 5,000 elderly persons who participated in telephone interviews after being identified through more than 65,000 calls to residential listings. Subjects were persons aged 65 years and older who resided in 108 counties in western Texas. The response rate was 72%. The probability of seeing a physician in the last 6 months for urban and rural residents was modeled using multiple logistic regression analysis. Among rural residents, characteristics that were significantly (p < 0.05) associated with not recently visiting a physician when health status is held constant included belief in home remedies, having less than a high school education, lack of health insurance, and low income. Among urban residents, Hispanic ethnicity and skepticism about medical care were negatively associated with having a recent visit, whereas being religious was positively associated. Despite the availability of Medicare coverage, several subgroups of the elderly population have impaired access to medical care in this sparsely populated region. Intensified outreach efforts are indicated. PMID- 12585778 TI - Ancillary community pharmacy services provided to older people in a largely rural and ethnically diverse region: a survey of consumers in West Texas. AB - Little is known about the quality of pharmacy services provided to the rural elderly population. This exploratory study examines rural/urban and ethnic differences in perceived access to ancillary pharmacy services among elderly people. Two telephone surveys were conducted using directory listings in West Texas to generate a longitudinal sample. Persons aged 65 years and older who were not cognitively impaired were asked to complete the survey. The number of participants in both rounds of the survey was 3,689. Seven ancillary pharmacy services were examined: delivery of medications, medication counseling, written medication information, blood pressure monitoring, blood glucose monitoring, osteoporosis screening, and immunization. The sample was stratified by county of residence (urban, rural, or frontier) and racial/ethnic background. Chi-square tests were performed to detect rural/urban and racial/ethnic differences in access to the seven ancillary services. The association between proficiency in English and access to the services was also examined. Rural residents were more likely than urban residents to report that their pharmacies provide delivery of medications, medication counseling, and immunization services, but they were less likely than their urban counterparts to report that their pharmacies provide blood pressure monitoring. Access to ancillary pharmacy services was reported as poorer by older Hispanic people compared with non-Hispanics. Deficiency in English was significantly related to inequality in reported access to ancillary pharmacy services. It is essential to consider the special needs of rural and Hispanic elderly people to ensure equitable access to ancillary pharmacy services. PMID- 12585779 TI - State of nursing science: on the edge. AB - Nurse scientists have had great and important successes contributing to science and to patient care. To review the state of nursing science is an enormous and complex challenge, and yet the pace of discovery constantly quickens. The purpose of this article, which was read at the 2002 State of the Science Congress, was to use the human response model to describe the domains of nursing science and note exemplary, innovative developments in the context of the model's "person" and "environmental" domains. Advances are noted in genetics, aging, development, and gender studies; also noted is work in infection care, disaster care, and identification of health disparities. Asked to predict the future, we chose instead to describe societal challenges and speculate how nursing leaders can contribute importantly by applying nursing's unique perspective. Benchmarks of our success will include reversal of the nursing shortage, patients living to their highest potential, and penetration of evidence based care into clinical practice and health policy. Furthermore, the media and the public will view nurse scientists as key informants related to clinical care. Nurse scientists will be elected to lead major interdisciplinary organizations, our training programs will prepare new scientists with the knowledge and skills to enter a competitive and ever-evolving field, our schools will have adequate infrastructure to support the advancement using cutting-edge technology, centers of excellence will provide research consultation and collaboration beyond university boundaries, and nurse scientists will assume a more visible role in translational research. PMID- 12585780 TI - Commentary: The science of human health-domain structure and future vision. PMID- 12585781 TI - Effects of exercise on bone density, balance, and self-efficacy in older women. AB - The effects of weighted vest walking and strength-training exercises on bone mineral density (BMD), balance, strength, and self-efficacy were tested in older women. Eighteen women, age 69.2 +/- 3.5 years, were randomly assigned to an exercise group (EG) (n = 9), or a sedentary control group (CG) (n = 9). The EG participated in 32 weeks (three 1-h sessions/week) of supervised strength training and walking, stair climbing, and balance exercises while wearing weighted vests. The CG did not exercise. All women took Ca2+ and vitamin D during the study period. Measures included 1) BMD of the hip and lumbar spine measured by dual-energy X-ray absorptiometry, 2) strength, 3) balance, and 4) scores on a self-efficacy instrument. The EG had significant improvements in bone density of the femoral neck and balance and a significant weight loss (P < 0.05). There were no changes in self-efficacy in either group. PMID- 12585782 TI - Effects of an off-site walking program on energy expenditure, serum lipids, and glucose metabolism in middle-aged women. AB - The present study aims to identify the effects of systematic walking on exercise energy expenditure (EEE) and blood profiles in middle-aged women. Fifty-two female nurse managers, aged 32 to 57 years (42.0 +/- 6.2), were randomly assigned to an intervention group (IG) and a control group (CG) for a 12-week study of the walking program. EEE was measured using a microelectronic device. Blood profiles were assessed before and after the walking program. The mean EEE (kcal/kg/d) in the IG and CG was 4.73 +/- 1.02 and 3.88 +/- 0.81 (P = 0.01), indicating an increase of 1.17 +/- 0.98 and 0.46 +/- 0.68 from baseline (P = 0.01), respectively. The mean change in high-density lipoprotein cholesterol in the IG and CG was 1.8 +/- 8.3 mg/dL and -2.9 +/- 7.0 mg/dL (P = 0.051); that in insulin was -4.5 +/- 7.5 microU/dL and -0.6 +/- 4.3 microU/dL (P = 0.046), respectively. These results show that systematic walking increases EEE and improves blood profiles. PMID- 12585783 TI - Coping strategies and life satisfaction in subgrouped fibromyalgia patients. AB - The present study describes pain- and stress-coping strategies and life satisfaction in subgroups of fibromyalgia patients. Thirty-two females with fibromyalgia syndrome (FMS) and 21 healthy pain-free women were studied. Those with FMS were classified as thermal (both heat and cold) pain sensitive or slightly cold pain sensitive based on pain thresholds determined using a Thermotest device. Global stress-coping styles, life satisfaction, and specific pain-coping strategies were measured. Patients classified as thermal pain sensitive were affected by physical symptoms to a greater extent than were those classified as slightly cold pain sensitive. The thermal pain sensitive group used more diverting attention coping strategies than the slightly cold pain sensitive group did. Separating fibromyalgia patients into subgroups might increase the potential for improving nursing care of these patients. Through the use of effective coping strategies in dealing with stress and pain, life satisfaction may also be enhanced. PMID- 12585784 TI - Laboratory measures of alcohol (ethanol) consumption: strategies to assess drinking patterns with biochemical measures. AB - Alcohol (ethanol) use is a global, health-related problem that spans a continuum ranging from low-risk, at-risk, and problem drinking to alcohol dependence and chronic abuse. Clinicians and researchers alike have the need to quantify drinking patterns to determine the risk for adverse, health-related events such as injury, liver damage, and cancer Biochemical measures of ethanol consumption are affected by temporal patterns of drinking as well as individual characteristics such as gender and age. The choice of a laboratory analysis to determine ethanol consumption is complex; no single laboratory test will predict drinking accurately across all drinking patterns, across the life span, and across gender. In conjunction with interviews and physical assessment, however, biochemical laboratory tests are sensitive tools used to measure both recent and long-term patterns of alcohol consumption. PMID- 12585785 TI - A theoretical model of the role of brain stem nuclei in alcohol-mediated arrhythmogenesis in older adults. AB - Uncertainty about the mechanism of alcohol-mediated arrhythmogenesis and the effect of alcohol use on arrhythmic risk among older adults is an increasing concern in light of population aging and recent reports that moderate alcohol consumption may protect older adults against coronary artery disease. In this review, a theoretical model of the role of brain stem nuclei in alcohol-mediated arrhythmogenesis in older adults is developed. The model is based on the hypothesis that the effects of alcohol on central autonomic pathways of cardiac control may alter the threshold for alcohol-mediated arrhythmogenesis among older adults. Findings from multiple lines of research including cellular, electrophysiological, epidemiological, experimental, and clinical studies in human, animal, and in vitro models were synthesized in developing the model. Suggestions for future research on the topic of alcohol-mediated arrhythmogenesis in older adults are offered. PMID- 12585786 TI - Providers, outcomes and their determinants. PMID- 12585787 TI - Necrotizing fasciitis secondary to chickenpox infection in children. AB - BACKGROUND: Necrotizing fasciitis is an uncommon but serious complication of chickenpox infection in young children. Because many of these infections affect the musculoskeletal tissues, orthopedic surgeons are often the first caregivers to be involved in diagnosis and treatment. Our objective was to review the diagnostic features of necrotizing fasciitis and analyze treatment methods to control and eradicate the musculoskeletal infection. DESIGN: A review. SETTING: The Children's Hospital of Eastern Ontario, Ottawa, a major Canadian pediatric trauma and referral centre. PATIENTS: Five children who presented with necrotizing fasciitis secondary to chickenpox infection. INTERVENTION: Surgical debridement of the involved area of necrotizing fasciitis and intravenous antibiotic treatment with clindamycin and penicillin. MAIN OUTCOME MEASURES: Complications outcome. RESULTS: The average age of the 5 children at presentation was 3.8 years (range from 2.9-5.8 yr). The necrotizing fasciitis involved the lower extremity in 5 children, the upper extremity in 3, and the abdomen, chest, neck and back in 1 child each. One child presented with involvement of all 4 extremities. In 4 children, culture specimens grew group A beta-hemolytic Streptococcus. They all survived and all limbs were salvaged, although secondary closure and skin grafting were required. At an average follow-up of 1 year, each child had fully recovered with no loss of muscle function. CONCLUSIONS: Necrotizing fasciitis should be suspected in any child with a history of varicella infection and an increasing complaint of pain and swelling in an extremity or other body area associated with increasing fever, erythema, lethargy and irritability. Emergent surgical debridement and intensive antibiotic therapy are essential to prevent muscle necrosis, major limb dysfunction and death. PMID- 12585789 TI - SESAP question. An empyema that has reached the chronic or organized phase. PMID- 12585788 TI - Provider volume and other predictors of outcome after total knee arthroplasty: a population study in Ontario. AB - INTRODUCTION: Because of rationing of the limited pool of health care resources, access to total knee arthroplasty (TKA) is limited, but investigation of variables that predict complications, length of hospital stay, cost and outcomes of TKA may allow us to optimize the available resources. The objective of this study was to examine the effect of various factors on complication rates after TKA in patients managed in Ontario. METHODS: Patients who had undergone an elective TKA between 1993 and 1996, as captured in the Canadian Institute for Health Information (CIHI) database, formed the study cohort. The CIHI dataset was used to obtain information regarding in-hospital complications, hospital length of stay, revision rates, infection rates and mortality. Generalized estimating linear or logistic regression equations were used to model outcomes as a function of age, gender, comorbidity, diagnosis and provider volume. RESULTS: During the study period, 14,352 patients in Ontario underwent TKA. Mortality at 3 months was associated with patient age, gender and comorbidity. There was no association between provider volume and mortality or the infection rate. Higher revision rates at 1 and 3 years were significantly associated with lower patient age and low hospital volume (p < 0.05). Hospitals in which fewer than 48 TKA procedures were done per year (< 40th percentile) had 2.2-fold greater 1-year revision rates than hospitals performing more than 113 TKAs annually (> 80th percentile). Complications during admission were associated with increased patient age and comorbidity, and higher hospital volume. Longer hospital stay was associated with female gender, increasing patient comorbidity and age, and lower provider volume. Surgeons who performed fewer than 14 TKAs annually (< 40th percentile) kept patients in hospital an average of 1.4 days longer than surgeons performing more than 42 TKAs annually (> 80th percentile). CONCLUSIONS: Patient variables significantly affect the rate of complications. Age, sex and comorbidity were significant predictors of complications, length of hospital stay and mortality after TKA. Although low surgeon volume was related to longer hospital stay, there was no association between surgeon volume and complication rates. The increased early revision rate for low-volume hospitals demands further study. PMID- 12585790 TI - Priority setting in a Canadian surgical department: a case study using program budgeting and marginal analysis. AB - INTRODUCTION: A key mandate of Canadian regional health authorities is to set priorities and allocate resources within a limited finding envelope. The objective in this study was to determine how resources within a surgical program in a Canadian rural hospital might be reallocated to better meet the needs of the local community. METHODS: Early in 2001, at the Canmore General Hospital, Canmore, Alta., an expert-panel working group, consisting of a community health service leader, operating-room nurse clinician, acute care head nurse and a general surgeon, assisted by a research assistant and 2 health economists carried out a program budgeting and marginal analysis project to assess multiple data inputs into the decision-making process and to develop recommendations for service expansion and resource release. They considered the cost and benefits of altering the mix of resources used, based on Headwaters Health Authority activity and financial data, and local expert opinion. RESULTS: The primary recommendation was to implement an additional surgery day per week (38 days of major surgery and 12 days of minor surgery over a 50-week year). However, the total dollars to fund such an expansion could not be released from within the Canmore budget, and additional dollars were not forthcoming from the health region. A secondary objective of implementing an additional minor surgery day every 3 weeks was pursued and the required resources were obtained. CONCLUSIONS: Due to resource constraints in health care, efforts by both clinicians and administrators should be made to better spend available resources. The marginal analysis process used in this study served as a useful framework for priority setting, which is generalizable to other surgical and nonsurgical programs in Canada. PMID- 12585791 TI - Effect of adjunctive range-of-motion therapy after primary total knee arthroplasty on the use of health services after hospital discharge. AB - INTRODUCTION: There is controversy as to whether continuous passive motion (CPM) after total knee arthroplasty (TKA), which is the standard treatment, confers significant benefit with respect to outcome. The primary purpose of this study was to determine if CPM or slider-board (SB) therapy, used as adjuncts to standardized exercises (SEs) during the acute-care hospital stay, resulted in a reduced total length of hospitalization and post-discharge rehabilitation in patients who underwent primary TKA. METHODS: We carried out a randomized, clinical trial on 120 patients who received a TKA at the University of Alberta Hospital, Edmonton, a tertiary care institution. The study horizon began at the point of discharge from the hospital and continued up to 6 months after operation. Postoperatively, patients (40 in each group) received CPM and SEs, SB therapy and SEs or SEs alone while in the tertiary Health service use was compared using transfer institution length of stay(LOS), post-discharge rehabilitation, readmission and complication rates and their associated costs. RESULTS: There were no differences in health service use or costs among the 3 groups over the 6-month study. The rates of postoperative complications and readmissions also were similar among the groups. Increased health service use associated with knee flexion that was less than 60 degrees at discharge, but similar proportions of patients with poor knee range of movement (ROM) at discharge were found in each group. CONCLUSIONS: This finding suggests that adjunctive ROM therapy, as used in this study, does not reduce health service use. Further research is required to determine if adjunctive ROM therapy after discharge from the surgical hospital decreases health service utilization in those patients who have poor knee ROM at the time of discharge. PMID- 12585792 TI - Thymoma: a review of the clinical and pathological findings in 65 cases. AB - INTRODUCTION: Although rare, thymoma is the most common tumour of the anterior mediastinum. In an effort to assess the clinical and pathologic characteristics of this tumour and to determine whether clinicopathologic stage or histopathologic classification correlates with clinical outcome, in the Department of Pathology and the Department of Surgery at the University of Saskatchewan we reviewed all cases of thymoma registered in the province of Saskatchewan using the database of the Saskatchewan Cancer Centre. METHODS: In 65 patients with a diagnosis of thymoma or thymic carcinoma identified from the Saskatchewan Cancer Centre database between Jan. 1, 1960, and Dec. 31, 2000, we studied the presentation, diagnostic investigations, therapeutic interventions, tumour size, postoperative course, clinical stage, histopathologic classification, disease recurrence and mortality. RESULTS: Of the 65 patients, 17 (26%) were asymptomatic and 11 (17%) had symptoms consistent with myasthenia gravis. Surgical resection is most commonly performed through a median sternotomy and frequently requires en bloc resection of one or more adjacent structures. The overall survival of patients with thymomawas found to correlate with the clinical stage as described by Masaoka and colleagues and with complete tumour resection. A trend to clinicopathologic correlation was observed when applying the histologic classification systems of Suster and Moran and the World Health Organisation, but this trend was not statistically significant. CONCLUSIONS: Thymoma is a rare tumour with a variable clinical presentation. Clinical outcome correlates with clinical stage and the ability to achieve complete tumour resection. PMID- 12585793 TI - Elective lumbar spinal decompression in the elderly: is it a high-risk operation? AB - INTRODUCTION: Spinal degeneration leading to spinal stenosis is increasingly common in an aging population. Many patients are not referred for operation because of the fear of severe complications. The purpose of this study was to relate the surgical risks involved in elective spinal decompression in elderly patients to those of total hip arthroplasty, a well-accepted procedure whose risks and benefits are well known. METHODS: We reviewed the charts of 6 8 consecutive patients who underwent elective spinal decompression and fitted our inclusion criteria (65-80 yr of age, no spinal or hip operations, no hip fractures, spine fractures or cauda equina syndrome and no bone or metastatic disease). This group was matched with a similar group of 68 randomly selected patients who underwent total hip arthroplasty during the same period. We recorded the age, gender, American Society of Anesthesiologists (ASA) score, early postoperative complication rate, operative time, hospital stay and blood lost. RESULTS: The 2 groups were well matched with respect to age, gender and ASA score. The only significant intraoperative difference was operative time, with the spine procedure taking longer to complete (191 min v. 278 min). Blood loss was not significantly different. Both groups had a similar number of lift threatening complications (12 v. 14). The number of minor complications was greater in the spinal decompression group (62 v. 125). CONCLUSIONS: Elective lumbar spinal decompression in elderly patients suffering significant pain or disability due to degenerative lumbar spinal stenosis is worthwhile. We recommend judicious use of conservative measures, but encourage referral for surgical management when these fail. This operation, even when instrumentation and bone grafting are required, is not associated with more serious or life-threatening risks than a total hip replacement in elderly patients. PMID- 12585794 TI - Musculoskeletal images. Chronic osteomyelitis. PMID- 12585796 TI - Musculoskeletal case 26. Osteoid osteoma. PMID- 12585795 TI - Soft-tissue images. "Phrygian cap" gallbladder. PMID- 12585798 TI - Canadian Association of General Surgeons Evidence Based Reviews in Surgery. 5. Need for preoperative radiation in rectal cancer. Preoperative radiotherapy combined with total mesorectal excision for resectable rectal cancer. PMID- 12585800 TI - East and Central African Journal of Surgery: another Canadian connection. PMID- 12585797 TI - Soft-tissue case 49. Cecal diverticulitis. PMID- 12585799 TI - Pharyngeal perforation caused by blunt trauma to the neck. PMID- 12585801 TI - Complete slipping of the capital femoral epiphysis after hematogenous osteomyelitis. PMID- 12585802 TI - Medical management of osteoid osteoma. PMID- 12585803 TI - The minority slowness effect: subtle inhibitions in the expression of views not shared by others. AB - Five studies revealed that people who hold the minority opinion express that opinion less quickly than people who hold the majority opinion. The difference in speed in the expression of the minority and majority opinions grew as the difference in the size of the minority and majority grew. Also, those with the minority view were particularly slow when they assumed the majority to be large, whereas the opposite was true for those with the majority view. The minority slowness effect was not found to be linked to attitude strength, nor was it influenced by anticipated public disclosure of the attitude. The effect is discussed in the context of implicit conformity pressures and the limited buffering effect of false consensus assumptions. PMID- 12585804 TI - Stereotypes as dominant responses: on the "social facilitation" of prejudice in anticipated public contexts. AB - This article challenges the highly intuitive assumption that prejudice should be less likely in public compared with private settings. It proposes that stereotypes may be conceptualized as a type of dominant response (C. L. Hull, 1943; R. B. Zajonc, 1965) whose expression may be enhanced in public settings, especially among individuals high in social anxiety. Support was found for this framework in an impression formation paradigm (Experiment 1) and in a speeded task designed to measure stereotypic errors in perceptual identification (Experiment 2). Use of the process dissociation procedure (B. K. Payne, L. L. Jacoby, & A. J. Lambert, in press) demonstrated that these effects were due to decreases in cognitive control rather than increases in stereotype accessibility. The findings highlight a heretofore unknown and ironic consequence of anticipated public settings: Warning people that others may be privy to their responses may actually increase prejudice among the very people who are most worried about doing the wrong thing in public. PMID- 12585805 TI - Leading us not unto temptation: momentary allurements elicit overriding goal activation. AB - The present research explored the nature of automatic associations formed between short-term motives (temptations) and the overriding goals with which they interfere. Five experimental studies, encompassing several self-regulatory domains, found that temptations tend to activate such higher priority goals, whereas the latter tend to inhibit the temptations. These activation patterns occurred outside of participants' conscious awareness and did not appear to tax their mental resources. Moreover, they varied as a function of subjective goal importance and were more pronounced for successful versus unsuccessful self regulators in a given domain. Finally, priming by temptation stimuli was found not only to influence the activation of overriding goals but also to affect goal congruent behavioral choices. PMID- 12585806 TI - Blood is thicker than water: kinship orientation across adulthood. AB - The importance of kin relationships was investigated across adulthood with 5 samples (total N = 1,365). Within the personal networks, the genetic relatedness with relationship partners predicted subjective closeness (mean r = .50) and social support (mean r = .13). Effects were robust in 2 samples when controlling for residential proximity and contact frequency. These intraindividual correlations showed considerable variability and were interpreted as individual expressions of nepotism. The heritability of individual nepotism was zero. Variability of nepotism was unrelated to personality traits, but substantially related to sex, and parental and partner status. The authors discuss subjective closeness as 1 proximate cue to kinship, and suggest nepotistic adaptations as powerful mechanisms in social relationships. PMID- 12585807 TI - Preference-consistent evaluation of information in the hidden profile paradigm: beyond group-level explanations for the dominance of shared information in group decisions. AB - Common explanations for the failure of groups to solve so-called hidden profiles focus on group processes, namely insufficient discussion of unshared information and premature consensus on a suboptimal alternative. As 2 experiments show, even in the absence of such group processes, hidden profiles are hardly ever solved. In Experiment 1, participants first received individual information about a personnel selection task and then read a group discussion protocol containing full information exchange. If the individual information was misleading (hidden profile), most participants failed to detect the correct alternative. In Experiment 2, it was determined that this effect is due to preference-consistent evaluation of information that constitutes an individual-level process mediating the failure of group members to solve hidden profiles. PMID- 12585808 TI - Individual and structural origins of friendship and social position among professionals. AB - Friendship affects individual and organizational well-being through direct relations, social positions, and complex network structures. In this study, the authors use longitudinal data from 2 groups of master's of business administration students to increase understanding of how friendship networks develop. The authors propose and test a dynamic model in which attribute similarity facilitates dyadic friendship ties, as well as similar network centrality and social position; early friendship increases later similarity in structural position and centrality; and early structural similarity enhances the likelihood of future friendship. Findings largely supported the model, demonstrating how homophily and early social contacts can jointly shape maturing friendship networks. PMID- 12585809 TI - Was it a phase? Young women's relinquishment of lesbian/bisexual identities over a 5-year period. AB - There have been many anecdotal accounts of individuals who self-identify as lesbian, gay, or bisexual only to relinquish these identities later. The current study examines this phenomenon among a sample of young nonheterosexual women who underwent 3 interviews over a 5-year period. Over a quarter of these women relinquished their lesbian/bisexual identities during this period: half reclaimed heterosexual identities and half gave up all identity labels. These women did not differ from those who maintained lesbian/bisexual identities regarding the age at which they underwent sexual identity milestones, the factors that precipitated their sexual questioning, or their recollection of childhood "indicators" of same sex sexuality. Women who relinquished their identities for heterosexual identities had smaller ratios of same-sex to other-sex attractions across the 5 year assessment period, but their attractions did not significantly change. Only 1 woman described her previous same-sex identification as a phase; the rest emphasized changes in how they interpreted or acted on their attractions. PMID- 12585810 TI - What good are positive emotions in crises? A prospective study of resilience and emotions following the terrorist attacks on the United States on September 11th, 2001. AB - Extrapolating from B. L. Fredrickson's (1998, 2001) broaden-and-build theory of positive emotions, the authors hypothesized that positive emotions are active ingredients within trait resilience. U.S. college students (18 men and 28 women) were tested in early 2001 and again in the weeks following the September 11th terrorist attacks. Mediational analyses showed that positive emotions experienced in the wake of the attacks--gratitude, interest, love, and so forth--fully accounted for the relations between (a) precrisis resilience and later development of depressive symptoms and (b) precrisis resilience and postcrisis growth in psychological resources. Findings suggest that positive emotions in the aftermath of crises buffer resilient people against depression and fuel thriving, consistent with the broaden-and-build theory. Discussion touches on implications for coping. PMID- 12585811 TI - Counting blessings versus burdens: an experimental investigation of gratitude and subjective well-being in daily life. AB - The effect of a grateful outlook on psychological and physical well-being was examined. In Studies 1 and 2, participants were randomly assigned to 1 of 3 experimental conditions (hassles, gratitude listing, and either neutral life events or social comparison); they then kept weekly (Study 1) or daily (Study 2) records of their moods, coping behaviors, health behaviors, physical symptoms, and overall life appraisals. In a 3rd study, persons with neuromuscular disease were randomly assigned to either the gratitude condition or to a control condition. The gratitude-outlook groups exhibited heightened well-being across several, though not all, of the outcome measures across the 3 studies, relative to the comparison groups. The effect on positive affect appeared to be the most robust finding. Results suggest that a conscious focus on blessings may have emotional and interpersonal benefits. PMID- 12585812 TI - Personality and the predisposition to engage in risky or problem behaviors during adolescence. AB - Longitudinal data from a representative sample of 1,978 Black and White adolescents were used to examine the role of personality in multiple risk or problem behaviors. Results indicate that covariation among diverse behaviors (educational underachievement, delinquent behavior, substance use, sexual behavior) can be adequately modeled by a single higher order factor, and that impulsivity and avoidance coping serve as generalized risk factors for involvement in these behaviors. Whereas none of the personality variables examined explained change in problem behaviors once established, avoidance coping prospectively predicted involvement among a subgroup of adolescents with little or no prior involvement. Results suggest that dysfunctional styles of regulating emotions and emotionally driven behaviors are core features of risky or problem behaviors during adolescence. PMID- 12585813 TI - Big Five factors of personality and replicated predictions of behavior. AB - Measures of the Big Five factors of personality were used to predict a variety of criterion variables thought to represent behaviors of some social and cultural significance (e.g., alcohol consumption, grade point average). Analyses focused on replicated predictions across 2 independent samples of participants (Ns = 276 and 142) with 3 different measures of the Big Five (the NEO Five-Factor Inventory, the Revised NEO Personality Inventory, and the Five-Factor Nonverbal Personality Questionnaire, the latter an experimental nonverbal personality inventory). The results indicated substantial consistency in behavior predictions across the different Big Five assessments. The data are interpreted as supporting both the construct validity of the personality measures used and the role of the Big Five factors as determinants of certain complex behaviors. PMID- 12585814 TI - Cognitive and motivational processes underlying coping flexibility: a dual process model. AB - Discriminative facility was proposed as a cognitive process and need for closure was proposed as a motivational process underlying coping flexibility. The dual process model posits that need for closure influences discriminative facility, which in turn modifies coping flexibility and psychological adjustment. In Study 1, results of structural equation modeling provided support for the dual-process model. This model was further examined using experimental methods (Study 2) and a prospective design (Study 3). Consistent with the dual-process model, results from all 3 studies showed that participants who were more motivated to seek alternative coping strategies tended to encode stressful situations in a more differentiated way. These individuals used a greater variety of strategies to fit different situational demands and were better adjusted. PMID- 12585815 TI - Determinants of health knowledge: an investigation of age, gender, abilities, personality, and interests. AB - Ten areas of health knowledge were investigated in 2 studies, 1 of college students (N = 169) and 1 of adults from the community (ages 19-70; N = 176). Measures assessed knowledge of aging, orthopedic/dermatological concerns, common illnesses, childhood/early life, serious illnesses, mental health, nutrition, reproduction, safety, and treatment of illness/disease. Significant gender differences favoring women were found for most areas of health knowledge, especially reproduction and early life. Results showed that cognitive ability accounted for the most variance in health knowledge with nonability (personality and interest traits) and demographic variables accounting for smaller but significant amounts of variance across most knowledge domains. PMID- 12585816 TI - Information processing in autistic children: more sequential or more simultaneous? AB - The objective of this study was to refine the understanding of cognitive troubles in autism and to clarify the modes of information processing mobilized by the subjects. Eight children were examined, aged 4;9-9;1, with a diagnosis of moderate autism. The Kaufman Assessment Battery for Children (K-ABC) was presented to each child. The results support an executive functioning deficit interpretation and a weak drive for central coherence in autism. Practical implications of this study in terms of follow up are also discussed. PMID- 12585817 TI - Do high functioning autistic individuals treated in a residential setting differ in divided attention abilities from those treated in an out-patient setting? AB - The goal of this current study was to test divided attention abilities of a group adults with autism and normal intellectual functioning, treated in a residential setting versus those treated in an out-patient setting. Both groups were compared with a control group using the Sternberg (1969) reaction time paradigm. It appeared that the in-patient group suffered from a divided attention deficit compared to a norm group. The patients treated in an out-patient setting scored in between the norm group and the in-patient group. Findings are discussed with respect to the relevance for the day to day clinical practice. PMID- 12585819 TI - Coping strategies and parental attitudes, a comparison of parents with children with autistic spectrum disorders and parents with non-autistic children. AB - This study focused on the coping strategies of parents' with children with autistic spectrum disorders (ASD) and the relation between these strategies and parenting styles. Coping strategies were measured using the Sense of Coherence Scale (SOC) and the Purpose in Life Test (PIL-R). Parental attitudes toward loving care, stress, worry, and guilt feelings were assessed using the Family Impact Questionnaire. Two groups of participants were included: parents with children with ASD (EG) (n = 66) and a matched control group (CG) (n = 66). Paired Samples t-Test and Pearson's r correlation were used as methods of analysis. Main results distinguished significant (p < .001 to .003) differences between the EG and CG for almost all variables included. The M level of coping strategy was much higher for the CG than for the EG. SOC showed a stress-reducing effect in both the EG and CG. PIL-R explained 50% of the variance in SOC for the EG and 33% for the CG. The only significant gender difference in the EG was on SOC indicating a higher sense of coherence among the fathers and probably an indicator of a stronger burnout effect of the mothers. PMID- 12585818 TI - A quantitative controlled MRI study of the brain in 28 persons with Asperger syndrome. AB - BACKGROUND: As structural brain abnormalities have been reported in infantile autism, the aim of this study was to determine whether such findings also exist in Asperger Syndrome (AS). METHODS: The diagnosis of Asperger Syndrome was based on the criteria in ICD-10 and DSM-IV. Brain magnetic resonance imaging (MRI) was performed with a 1.5 T imager. T2-weighted axial and coronal slices and T1 weighted three dimensional sagittal slices were obtained and visual and quantitative analysis were performed. SUBJECTS: There were 28 Asperger individuals, 17 children and adolescents (age 6-19 years, mean 12.4 years), 11 adults (age 20-60 years, mean 37. 9 years) and 28 healthy age and gender matched controls. RESULTS: Mild inconsistent alterations were detected in 13/28 of the individuals with Asperger Syndrome compared to 6/23 in the comparable controls. There were no differences between the right and left hemispheres, nor was there any abnormality in terms of myelination or migration. The anterior-posterior diameters of the mesencephalon were statistically significantly shorter in the Asperger syndrome individuals than in the controls. CONCLUSIONS: No consistent focal brain abnormalities for Asperger Syndrome were detected. The reduced diameters of the mesencephalon in the Asperger group support the hypothesis that the mesencephalon may be involved in the pathogenesis of Asperger Syndrome. PMID- 12585820 TI - Children and adolescents with developmental disorders and violence. AB - OBJECTIVES: The aim of this preliminary study was to investigate how commonly child-psychiatric inpatients have experienced and seen violence, and whether children with developmental disorders are at an increased risk for such traumatic events. METHODS: The sample consisted of 41 patients, 29 boys and 12 girls, with a mean (SD) age of 11.8 (3.2) years. Ten children had a developmental disorder, 19 an emotional disorder, 9 a behavioural disorder and 3 a psychotic disorder. The study was performed as part of the normal clinical examinations by paying special attention to violence in the lives of these patients. RESULTS: 88% of the patients had experienced some form of violence. 49% had experienced active physical violence and 49% active psychological violence. If suspected violence was also taken into account, active physical violence was more common among the patients with developmental disorders (90%) or behavioural disorders (78%) than among those with emotional disorders (37%) or psychotic disorders (67%). On the other hand, passive physical violence without active physical violence had been experienced by 37% of the patients with emotional disorders but not by the other patients. CONCLUSIONS: Violence was common among all groups of child-psychiatric inpatients. These individuals may be vulnerable to violence, as interaction with them may be especially demanding for peers and adults. PMID- 12585821 TI - The development and evaluation of a computer-based program to test and to teach the recognition of facial affect. AB - Autism is a chronic pervasive neurodevelopmental disorder characterized by the early onset of social and communicative impairments as well as restricted, ritualized, stereotypic behavior. The endophenotype of autism includes neuropsychological deficits, for instance a lack of "Theory of Mind" and problems recognizing facial affect. In this study, we report the development and evaluation of a computer-based program to teach and test the ability to identify basic facially expressed emotions. 10 adolescent or adult subjects with high functioning autism or Asperger-syndrome were included in the investigation. A priori the facial affect recognition test had shown good psychometric properties in a normative sample (internal consistency: rtt=.91-.95; retest reliability: rtt=.89-.92). In a prepost design, one half of the sample was randomly assigned to receive computer treatment while the other half of the sample served as control group. The training was conducted for five weeks, consisting of two hours training a week. The trained individuals improved significantly on the affect recognition task, but not on any other measure. Results support the usefulness of the program to teach the detection of facial affect. However, the improvement found is limited to a circumscribed area of social-communicative function and generalization is not ensured. PMID- 12585822 TI - Some aspects of treatment and habilitation of children and adolescents with autistic disorder in Northern-Finland. AB - Hospital records and data on the treatment/habilitation status of 187 children with autism aged 3-18 years were gathered from Northern Finland. The treatment programs and therapies varied, depending on the trained staff available. One hundred and fifty-two (82.9%) children and adolescents with autism received more than one therapeutic intervention or specific training program. The most common therapies were physiotherapy as well as speech, occupational and music therapy. 43.9% of the children and adolescents with autism received specific training according to TEACCH (Treatment and Education of Autistic and related Communication-Handicapped Children), 10.2% according to Lovaas and 30.5% according to the Portage program. Antiepileptic medication had been prescribed to 23.9% and psychopharmacological interventions to 14.9% of the individuals with autistic disorder (AD). One hundred and seventy-eight subjects out of 187 showed some improvement on the Childhood Autism Rating Scale (CARS), even if no statistically significant difference was found between the outcome of the available habilitation methods. PMID- 12585823 TI - Asperger syndrome with comorbid emotional disorder--treatment with psychoanalytic psychotherapy. AB - The purpose of psychoanalytic psychotherapy is to help the child to grow and develop normally. Children who are able to play, process their problems in the play. The therapist's role is to engage with the play and verbalize the emotions and the interactions demonstrated in it. This case-presentation illustrates how the child can express and work his inner fears and anxieties in play-situations with the therapist and shows how this interaction helps the child to develop and individuate. Children diagnosed as having Asperger syndrome may have emotional problems, in which case psychoanalytical psychotherapy is one of the means of helping the child and the family. PMID- 12585824 TI - Children with Asperger syndrome: specific aspects of their drawings. AB - Free drawings of children with AS, aged 7-16 years, were analysed in relation to the clinical picture comprising their difficulties in communication, social behaviour and cognition. All children showed good abilities in drawing. Pictures had some common traits and were distinctly original, reflecting peculiarities of the syndrome features. Analysis of free drawings was found to be a helpful tool in understanding the inner world and the dynamic changes during the therapy process of these children. PMID- 12585825 TI - The implications of regional variations in Medicare spending. Part 1: the content, quality, and accessibility of care. AB - BACKGROUND: The health implications of regional differences in Medicare spending are unknown. OBJECTIVE: To determine whether regions with higher Medicare spending provide better care. DESIGN: Cohort study. SETTING: National study of Medicare beneficiaries. PATIENTS: Patients hospitalized between 1993 and 1995 for hip fracture (n = 614,503), colorectal cancer (n = 195,429), or acute myocardial infarction (n = 159,393) and a representative sample (n = 18,190) drawn from the Medicare Current Beneficiary Survey (1992-1995). EXPOSURE MEASUREMENT: End-of life spending reflects the component of regional variation in Medicare spending that is unrelated to regional differences in illness. Each cohort member's exposure to different levels of spending was therefore defined by the level of end-of-life spending in his or her hospital referral region of residence (n = 306). OUTCOME MEASUREMENTS: Content of care (for example, frequency and type of services received), quality of care (for example, use of aspirin after acute myocardial infarction, influenza immunization), and access to care (for example, having a usual source of care). RESULTS: Average baseline health status of cohort members was similar across regions of differing spending levels, but patients in higher-spending regions received approximately 60% more care. The increased utilization was explained by more frequent physician visits, especially in the inpatient setting (rate ratios in the highest vs. the lowest quintile of hospital referral regions were 2.13 [95% CI, 2.12 to 2.14] for inpatient visits and 2.36 [CI, 2.33 to 2.39] for new inpatient consultations), more frequent tests and minor (but not major) procedures, and increased use of specialists and hospitals (rate ratio in the highest vs. the lowest quintile was 1.52 [CI, 1.50 to 1.54] for inpatient days and 1.55 [CI, 1.50 to 1.60] for intensive care unit days). Quality of care in higher-spending regions was no better on most measures and was worse for several preventive care measures. Access to care in higher-spending regions was also no better or worse. CONCLUSIONS: Regional differences in Medicare spending are largely explained by the more inpatient-based and specialist-oriented pattern of practice observed in high-spending regions. Neither quality of care nor access to care appear to be better for Medicare enrollees in higher-spending regions. PMID- 12585826 TI - The implications of regional variations in Medicare spending. Part 2: health outcomes and satisfaction with care. AB - BACKGROUND: The health implications of regional differences in Medicare spending are unknown. OBJECTIVE: To determine whether regions with higher Medicare spending achieve better survival, functional status, or satisfaction with care. DESIGN: Cohort study. SETTING: National study of Medicare beneficiaries. PATIENTS: Patients hospitalized between 1993 and 1995 for hip fracture (n = 614,503), colorectal cancer (n = 195,429), or acute myocardial infarction (n = 159,393) and a representative sample (n = 18,190) drawn from the Medicare Current Beneficiary Survey (MCBS) (1992-1995). EXPOSURE MEASUREMENT: End-of-life spending reflects the component of regional variation in Medicare spending that is unrelated to regional differences in illness. Each cohort member's exposure to different levels of spending was therefore defined by the level of end-of-life spending in his or her hospital referral region of residence (n = 306). OUTCOME MEASUREMENTS: 5-year mortality rate (all four cohorts), change in functional status (MCBS cohort), and satisfaction (MCBS cohort). RESULTS: Cohort members were similar in baseline health status, but those in regions with higher end-of life spending received 60% more care. Each 10% increase in regional end-of-life spending was associated with the following relative risks for death: hip fracture cohort, 1.003 (95% CI, 0.999 to 1.006); colorectal cancer cohort, 1.012 (CI, 1.004 to 1.019); acute myocardial infarction cohort, 1.007 (CI, 1.001 to 1.014); and MCBS cohort, 1.01 (CI, 0.99 to 1.03). There were no differences in the rate of decline in functional status across spending levels and no consistent differences in satisfaction. CONCLUSIONS: Medicare enrollees in higher-spending regions receive more care than those in lower-spending regions but do not have better health outcomes or satisfaction with care. Efforts to reduce spending should proceed with caution, but policies to better manage further spending growth are warranted. PMID- 12585827 TI - Interferon therapy after tumor ablation improves prognosis in patients with hepatocellular carcinoma associated with hepatitis C virus. AB - BACKGROUND: Even after the surgical or medical treatment of hepatocellular carcinoma, tumors frequently develop at new foci, leading to a poor prognosis. OBJECTIVE: To assess whether combined tumor ablation and interferon therapy can reduce the occurrence of new foci of hepatocellular carcinoma, thereby improving survival rate. DESIGN: Randomized, controlled study. SETTING: University hospital. PATIENTS: 74 patients with compensated cirrhosis, three or fewer nodules of hepatocellular carcinoma, and low hepatitis C virus RNA loads (< or =2 x 10(6) copies/mL). INTERVENTION: After all patients had complete ablation of lesions by percutaneous ethanol injection therapy, 49 patients were assigned to receive 6 million U of interferon three times weekly for 48 weeks and 25 did not receive treatment. MEASUREMENTS: Abdominal ultrasonography, computed tomography, and determination of blood biochemical measures. RESULTS: Of the 49 patients treated with interferon, 21 showed a sustained biochemical response and 14 showed a sustained virologic response. The rate of first recurrence of new foci of hepatocellular carcinoma was similar in patients treated with interferon and untreated patients; however, the rates of second or third recurrence seemed to be lower in the interferon group than in the untreated group. Patients treated with interferon had a survival rate of 68% at 5 years and 53% at 7 years; untreated patients had a survival rate of 48% at 5 years and 23% at 7 years. CONCLUSION: After tumor ablation by ethanol injection, interferon therapy may enhance patient survival in selected patients with chronic hepatitis C. PMID- 12585828 TI - Single-detector helical computed tomography as the primary diagnostic test in suspected pulmonary embolism: a multicenter clinical management study of 510 patients. AB - BACKGROUND: Helical computed tomography (CT) is a readily available tool for diagnosing pulmonary embolism (PE); however, its role in the management of patients with clinically suspected PE has not been fully established. OBJECTIVE: To determine the effectiveness and safety of using helical CT of the pulmonary arteries as the primary diagnostic test in patients with suspected PE. DESIGN: Multicenter, prospective clinical outcome study. SETTING: Two academic hospitals and one large teaching hospital in the Netherlands. PATIENTS: 510 consecutive inpatients and outpatients with clinically suspected PE followed for 3 months. INTERVENTIONS: Patients underwent helical CT of the pulmonary arteries within 24 hours after presenting with signs and symptoms of PE. If CT results were normal or inconclusive, compression ultrasonography was performed on the same day as CT and repeated on days 4 and 7 if findings on the first compression ultrasonography were normal. When CT or compression ultrasonography results were positive for thromboembolism, anticoagulation was started. Anticoagulation was not started when results of CT were negative for PE or indicated an alternative diagnosis that explained the clinical signs and symptoms, or when results on serial compression ultrasonography were normal. MEASUREMENTS: Patients received instructions to report any symptoms or signs of PE or deep venous thrombosis (DVT) during the 3-month follow-up period. The authors performed compression ultrasonography or phlebography for suspected DVT and pulmonary angiography for suspected PE. RESULTS: Computed tomography identified PE in 124 of 510 patients (24.3%) and an alternative diagnosis in 130 patients (25.5%); CT scans were normal in 248 patients (48.6%). The CT scan could not be interpreted in 8 patients (1.6%) and was not obtained in 2. Compression ultrasonography revealed DVT in 2 patients at the first examination; findings on repeated compression ultrasonography at days 4 and 7 were normal. Mortality in the patients with normal helical CT scans was 4.1% (10 of 246 patients). No patients in this group died of fatal PE, 1 patient developed nonfatal PE, and venous thromboembolism occurred in 0.4% of these patients (95% CI, 0% to 2.2%). In the patients with alternative diagnoses, 1 patient had DVT on objective testing during follow-up. Mortality in this group was 21.5% (28 of 130 patients); in 1 of these patients, PE could not be confidently ruled out as a contributing cause of death. Venous thromboembolism occurred in 1.5% of these patients (CI, 0.2% to 5.6%). CONCLUSIONS: In patients with suspected PE, helical CT can be used safely as the primary diagnostic test to rule out PE. Serial compression ultrasonography has limited additional value. PMID- 12585830 TI - Update in pulmonary medicine. PMID- 12585829 TI - Natural history of benign solid and cystic thyroid nodules. AB - BACKGROUND: Thyroid nodules are common and most often benign. The natural history of benign thyroid nodules, however, is unclear. OBJECTIVE: To determine the natural history of cytologically benign thyroid nodules using ultrasonography. DESIGN: Retrospective case series. SETTING: Single tertiary care clinic. PARTICIPANTS: All patients referred to the Brigham and Women's Hospital Thyroid Nodule Clinic, Boston, Massachusetts, who had benign cytologic results on ultrasonography-guided fine-needle aspiration of a thyroid nodule between 1995 and 2000 and returned for a requested follow-up examination 1 month to 5 years later. MEASUREMENTS: Nodule dimensions were measured at both visits, and growth was defined as an increase in calculated volume of 15% or greater. These results were correlated with the time between examinations, age, sex, baseline serum thyroid-stimulating hormone concentration, and cystic content of each nodule. RESULTS: Nodule volume increased over time (P < 0.001). The estimated proportion of nodules with an increase in volume of 15% or greater after 5 years was 89%. Nodules with greater cystic content were less likely to grow than solid nodules (P = 0.01). Seventy-four of the 330 nodules were reaspirated on the second visit. Despite an average increase in volume of 69%, only 1 of 74 reaspirated nodules was malignant. CONCLUSION: Most solid, benign thyroid nodules grow. Therefore, an increase in nodule volume alone is not a reliable predictor of malignancy. PMID- 12585831 TI - Managing the patient with venous ulcers. AB - Venous disease is the most common cause of leg ulcers. The refractory nature of venous ulcers affects the quality of life and work productivity of those persons afflicted. This, in combination with the high costs of long-term therapy, makes venous ulcers a major health problem in developed countries. Management of venous leg ulcers is based on understanding pathophysiologic abnormalities. In recent years, identifying prognostic factors for healing and developing novel therapeutic approaches for venous ulcers have offered valuable tools for the management of patients with this disorder. PMID- 12585832 TI - Hospice benefits and phase I cancer trials. AB - Medicare denies hospice coverage to patients with terminal illnesses who enroll as participants in phase I studies, which assess the toxicity and dosing of potential treatments for incurable diseases. Federal regulations require patients to forgo curative therapies, and they interpret phase I agents as treatment for the terminal condition for which hospice care was elected. Thus, by enrolling as a participant in a phase I trial, a patient otherwise eligible for hospice is rendered ineligible. Private insurers have similar provisions for children and adults younger than 65 years of age. Such exclusions are not defensible on ethical or clinical grounds. Policymakers, insurers, and institutional review boards all have a role in resolving this problem. PMID- 12585833 TI - Fabry disease, an under-recognized multisystemic disorder: expert recommendations for diagnosis, management, and enzyme replacement therapy. AB - Fabry disease (alpha-galactosidase A deficiency) is an X-linked recessive lysosomal storage disorder. Although the disease presents in childhood and culminates in cardiac, cerebrovascular, and end-stage renal disease, diagnosis is often delayed or missed. This paper reviews the key signs and symptoms of Fabry disease and provides expert recommendations for diagnosis, follow-up, medical management, and the use of enzyme replacement therapy. Recommendations are based on reviews of the literature on Fabry disease, results of recent clinical trials, and expertise of the authors, all of whom have extensive clinical experience with Fabry disease and lysosomal storage disorders and represent subspecialties involved in treatment. All males and female carriers affected with Fabry disease should be followed closely, regardless of symptoms or treatment status. Clinical trials have shown that recombinant human alpha-galactosidase A replacement therapy--the only disease-specific therapy currently available for Fabry disease- is safe and can reverse substrate storage in the lysosome, the pathophysiologic basis of the disease. Enzyme replacement therapy in all males with Fabry disease (including those with end-stage renal disease) and female carriers with substantial disease manifestations should be initiated as early as possible. Additional experience is needed before more specific recommendations can be made on optimal dosing regimens for reversal; maintenance; and prevention of disease manifestations in affected males, symptomatic carrier females, children, and patients with compromised renal function. PMID- 12585834 TI - Geographical variations in Medicare spending. PMID- 12585835 TI - What's enough, what's too much? PMID- 12585836 TI - The implications of regional variations in Medicare--what does it mean for Medicare? PMID- 12585837 TI - Science, public health, and public awareness: lessons from the Women's Health Initiative. PMID- 12585838 TI - Valentine. PMID- 12585839 TI - Screening for colorectal cancer. PMID- 12585840 TI - Screening for colorectal cancer. PMID- 12585841 TI - Screening for colorectal cancer. PMID- 12585842 TI - Acute hepatocellular-cholestatic liver injury after olanzapine therapy. PMID- 12585844 TI - Furosemide challenge in patients with heart failure and adverse reactions to sulfa-containing diuretics. PMID- 12585845 TI - Antral injections of botulinum a toxin for the treatment of obesity. PMID- 12585846 TI - Food and Drug Administration approval of buprenorphine-naloxone for office treatment of addiction. PMID- 12585848 TI - Menopausal hormone therapy: summary of a scientific workshop. PMID- 12585849 TI - Summaries for patients. Are thyroid nodules that grow cancerous? PMID- 12585850 TI - Summaries for patients. Detecting clots in the lungs. PMID- 12585851 TI - Summaries for patients. Interferon therapy improves survival in patients with liver cancer and hepatitis C virus infection. PMID- 12585852 TI - Summaries for patients. The implications of regional variations in Medicare spending: health outcomes and satisfaction with care. PMID- 12585853 TI - Summaries for patients. The implications of regional variations in Medicare spending: the content, quality, and accessibility of care. PMID- 12585854 TI - On the nature of the 'heterogeneous' catalyst: nickel-on-charcoal. AB - Results from aromatic aminations and Kumada couplings, together with spectroscopic analyses (TEM, EDX, ICP-AES, React-IR), reveal that catalysis using nickel-on-charcoal (Ni/C) is most likely of a homogeneous rather than heterogeneous nature. In the course of a reaction with Ni/C, nickel bleed from the support was calculated to be as high as 78%. However, the existence of an equilibrium for this homogeneous species between nickel located inside vs outside the pore system of charcoal strongly favors the former, thus leaving only traces of metal detectable in solution. This accounts for virtually complete recovery of nickel on the charcoal following filtration of a reaction mixture and allows for recycling of the catalyst. TEM and EDX data were used to explain different reactivity profiles of Ni/C, which depended upon the method of reduction used to convert Ni(II)/C to Ni(0) as well as the level of nickel loading on the support. PMID- 12585855 TI - Nickel-on-charcoal-catalyzed aromatic aminations and Kumada couplings: mechanistic and synthetic aspects. AB - Protocols for aromatic aminations and Kumada couplings catalyzed by 'heterogeneous' nickel-on-charcoal (Ni/C) have been revised, making them simpler and more time efficient. For both types of reactions, reduction of the catalyst precursor Ni(II)/C using n-BuLi prior to addition of a substrate can be avoided. Instead, in amination reactions, the amine in combination with lithium tert butoxide was found to convert Ni(II)/C to active Ni(0). For Kumada couplings, direct reduction of Ni(II)/C by the Grignard reagent is easily achieved. Reactions run in the presence of triarylphosphine ligands of varying substitution patterns and with differing electronic properties provided insight into the mechanism of these nickel-catalyzed transformations. Ligandless Kumada couplings were facile with aryl Grignards, which may be a consequence of pi-complexation of nickel by the aryl group in the reagent. Larger scale reactions of both types of couplings have been successfully performed, suggesting that Ni/C-based processes can be scaled-up as needed. PMID- 12585856 TI - Stereoselective alkylation of chiral 2-imidoylphenols with organolithium reagents: synthesis of enantiopure 2-aminoalkylphenols. AB - In this paper the addition of organolithium reagents to chiral imidoylphenols to prepare enantiopure phenolic Mannich-type bases is described. The experimental data show that this kind of imine is surprisingly reactive toward organolithium reagents, differently from classical imines, and does not need any Lewis acid or base activation. Moreover, interesting results have been obtained with aldimines but more unusually with ketimines. This reaction results in high yields and diastereoselectivities and allows the preparation of aminophenols quaternary at the C-1 carbon atom, which cannot be prepared with the methods available till now. The sense of asymmetric induction has been explained and confirmed in agreement with the results previously obtained by hydride reduction of the same substrates. In some cases this procedure is complementary to the reductive one, allowing the preparation of the diastereomers less abundant in the reduction. The reaction allows the synthesis of one or the other of the two diastereomers, choosing the opportune starting imidoylphenol and the organolithium reagent. PMID- 12585857 TI - Synthetic applications of lithiated N-Boc allylic amines as asymmetric homoenolate equivalents. AB - Lithiation of N-(Boc)-N-(p-methoxyphenyl) allylic amines in the presence of (-) sparteine provides asymmetric homoenolate equivalents which react with electrophiles to provide highly enantioenriched enecarbamates. Acidic hydrolysis of the enecarbamates can provide the corresponding beta-substituted aldehydes. A synthetic sequence that involves a stereocontrolled intramolecular nitrone-olefin dipolar cycloaddition has been developed for the preparation of enantioenriched 2 formyl-4-phenyl-1-aminocyclopentanes from one beta-allyl-substituted aldehyde. Further manipulations allow access to an enantioenriched beta-lactam. In another synthetic sequence, transmetalation of the lithiated intermediates and reactions with aldehyde electrophiles can be controlled to afford highly enantioenriched anti homoaldol products. Use of an anti aldehyde homoaldol product as the chiral electrophile in an iterative reaction provides a double homoaldol product containing four stereogenic centers with high diastereoselectivity and enantioselectivity. Reaction pathways are proposed to account for the observed products. PMID- 12585859 TI - Magnetic interaction of pyridyl-substituted thioaminyl stable free radicals. AB - N-(2-Pyridylthio)-2,6-diaryl-4-R-phenylaminyls (R = Ph, 4-ClC(6)H(4), MeCO, CN, EtOCO) and N-(4-pyridylthio)-2,6-diaryl-4-R-phenylaminyls (R = Ph, 4-ClC(6)H(4), EtOCO) were prepared and isolated as radical crystals. Their ESR spectra were measured, and the NS and pyridyl nitrogen and anilino meta and pyridyl ortho and para proton hyperfine coupling constants were determined. The spin-density calculations based on the density functional theory were performed by the UBecke 3LYP hybrid method using the STO 6-31G basis set. X-ray crystallographic analyses were performed for three radicals, and their structures were discussed in detail. The magnetic susceptibility measurements were carried out for the nine kinds of isolated radicals with a SQUID magnetometer. One radical showed ferromagnetic coupling (2J/k(B) = 44 K), and the others showed antiferromagnetic behavior. The magnetic interactions observed are explained on the basis of the crystal structures. PMID- 12585858 TI - First total synthesis of (+/-)-Linderol A, a tricyclic hexahydrodibenzofuran constituent of Lindera umbellata bark, with potent inhibitory activity on melanin biosynthesis of cultured B-16 melanoma cells. AB - The first total synthesis of (+/-)-Linderol A, a hexahydrodibenzofuran constituent of Lindera umbellata bark, with potent inhibitory activity on the melanin biosynthesis of cultured B-16 melanoma cells, was achieved through 19 steps of reaction in 6.6% overall yield, in which the critical step was a tandem reaction of a 3-ethoxycarbonylcoumarin derivative with dimethylsulfoxonium methylide to yield the 2-ethoxycarbonylcyclopenta[b]benzofuran-3-ol derivative. PMID- 12585860 TI - Synthesis of purine- and pyrimidine-substituted heptadienes. The stereochemistry of cyclization and cyclopolymerization products. AB - A series of 1,6-heptadienes, substituted in the 4 position with nucleic acid bases 1-6, have been synthesized via Mitsunobu condensations. Guanine, adenine, thymine, and uracil derivatives can be prepared directly by coupling the protected base with 1,6-heptadien-4-ol (7). However, coupling protected cytosine and 7 gives an O-alkylated product. Thus, the cytosine derivative must be prepared from the uracil-substituted heptadienes via the triazole. The free radical addition of CCl(4) and BrCCl(3) to these adducts was investigated. In all cases, both 1:1 and 1:2 adducts were obtained. The 1:1 adduct was identified as the cyclized product of the initially formed 5-hexen-1-yl radical. The cyclization takes place in a stereospecific manner, with only one of the four possible diastereomers resulting. NMR studies indicate that all substituents are cis in this product. In the case of the addition of CCl(4) to the uracil substituted heptadiene, this conclusion was confirmed by an X-ray structure determination of the isolated cyclized product. The free-radical-initiated cyclocopolymerizations of 1-6 with SO(2) gave 1:1 copolymers with cis-linked five membered rings. Two-dimensional NMR studies on poly(2-SO(2)) showed predominately the cis-syn isomer while poly(6-SO(2)) has an approximately equal amount of cis syn and cis-anti isomers. PMID- 12585861 TI - Synthesis of C-17-functionalized spongiane diterpenes: diastereoselective synthesis of (-)-spongian-16-oxo-17-al, (-)-acetyldendrillol-1, and (-) aplyroseol-14. AB - The diastereoselective synthesis of spongiane diterpenes (-)-spongian-16-oxo-17 al 2, (-)-acetyldendrillol-1 15, and (-)-aplyroseol-14 16 has been completed efficiently via the common intermediate 14. Compound 14 was prepared in five synthetic steps from (+)-podocarp-8(14)-en-13-one 13, easily available from commercial (-)-abietic acid. The key steps in the syntheses were a regioselective reduction of a 1,4-dialdehyde unit, a one-pot acetalization-acetylation, and a translactonization. The synthesis of 15 and 16 has led us to a revision of the configuration at C-17 for natural (-)-acetyldendrillol-1 and a structural reassignment for aplyroseol-14. Thus, aplyroseol-14 16 presents an unprecedented delta-lactone-based structure for spongiane-type diterpenoids. A theoretical study including a series of ab initio calculations for the mechanism involved in the conversion of ester 22 into natural product 2 has also been carried out. PMID- 12585862 TI - One-pot synthetic routes to multiply substituted indene derivatives by hydrolysis of zirconocene-mediated intermolecular coupling reactions of aromatic ketones and alkynes. AB - Two one-pot multicomponent synthetic methods for highly substituted indenes are described. The intermolecular coupling of aromatic ketones with alkynes on low valent zirconocene species generates oxazirconacyclopentenes, which upon hydrolysis with 20% HCl for 3 h afforded indene derivatives in good to excellent yields. Similarly, the pair-selective coupling of two identical or different alkynes bearing at least one aromatic substituent formed zirconacyclopentadienes. Quenching of the reaction mixture with concentrated H(2)SO(4) also results in the formation of highly substituted indenes in high yields. PMID- 12585863 TI - Modular approach to novel chiral aryl-ferrocenyl phosphines by Suzuki cross coupling. AB - Two novel planar chiral and atropisomeric P,N and P,O aryl-ferrocenyl ligand systems have been developed. The strategy is short and involves a new synthetic approach to aryl-ferrocenyl compounds via a Suzuki cross-coupling procedure. The modular design can easily give access to variety of chiral mono- and bidentate ligands. Two simple derivatives of a novel chiral bidenate P,N ligand belonging to the MOPF family have been synthesized and tested in the enantioselective copper-catalyzed addition of diethyl zinc to an enone and a "difficult" diester. Moderate to excellent yields and enantioselectivities up to 58% were obtained using 1 mol % Cu(OTf)(2) and 1.5 mol % chiral ligand. PMID- 12585864 TI - Pentamethylated and pentaphenylated azaferro- and azaruthenocenes: simple and general methodology for the preparation of enantiopure derivatives. AB - A methodology for the enantioselective synthesis of planar chiral 2-substituted 1',2',3',4',5'-pentamethylazaferro- and azaruthenocenes is reported. The key step is the introduction of an enantiopure chiral sulfoxide auxiliary via a selective ortho-lithiation with subsequent chromatographic separation of the resulting diastereomers. Cleaving off the sulfoxide auxiliary by treatment with t-BuLi generates an optically pure anion which may be quenched with a variety of electrophiles to give the optically pure azametallocene derivatives. In addition, it was attempted to extend the methodology to encompass 1',2',3',4',5' pentaphenyl derivatives. It was, however, not possible to attach the chiral sulfoxide auxiliary onto the pentaphenylated azaferrocene, and for the azaruthenocene case, only one diastereomer could be isolated. Most importantly, the sulfoxide group could be cleaved off and the resulting chiral azaruthenocenyl anion was quenched with paraformaldehyde and iodine, resulting in products with ee values of 85% and 99%, respectively. PMID- 12585865 TI - Novel synthesis of protected thiol end-capped stilbenes and oligo(phenylenevinylene)s (OPVs). AB - The first general procedures for preparation of thiol end-capped stilbenes and oligo(phenylenevinylene)s (OPVs) with tert-butyl- and acetyl-protected thiol termini have been developed. These reactions proceed via Br/Li exchange, McMurry, and Wittig-type reactions. The thiol functionality is protected against strong basic and acidic reaction conditions as a t-Bu sulfide. As a key point in the method, reprotection of the thiol group is accomplished by means of acetyl chloride and boron tribromide. The novel strategy forms the basis for stepwise introduction of 4-mercaptostyryl units in OPVs. The new mono-, di-, and trimercapto OPVs have potential applications as one, two, and three terminal molecular devices in gold nanoparticle clusters, self-assembled monolayers, and optoelectronic devices. PMID- 12585866 TI - Photochemical [2 + 2] cycloaddition of acetylene to chiral 2(5H)-furanones. AB - The [2 + 2] photocycloaddition of acetylene to chiral 2(5H)-furanones was investigated. The influence on the chemical yield and facial diastereoselectivity of the substituent at the stereogenic center and also the effect of a 4-methyl group were evaluated. A mechanistic proposal based on a simple theoretical conformational analysis is presented. Using a C(2)-symmetric bis(lactone) as the substrate, a diastereomeric excess higher than 98% was found. PMID- 12585867 TI - Influence of the correlation, aggregation, and solvation on ab initio computed Li C, Li-N, and Li-Li NMR coupling constants. AB - The (1)J and (3)J(C-Li), (1)J(N-Li), and (2)J(Li-Li) NMR coupling constants have been calculated for various homogeneous and heterogeneous aggregates of methyllithium and lithium dimethylamide at the HF and MP2 levels of calculation. Ethereal solvation has also been taken into account either through a continuum model or through the explicit introduction of Me(2)O molecules. The results obtained are in good general agreement with the experimental data available for methyllithium itself or model alkyllithiums and supports the empirical rule proposed by Bauer, Winchester, and Schleyer to evaluate (1)J(C-Li) provided that calculations include solvent and/or aggregation effects. PMID- 12585868 TI - A variable concept for the preparation of branched glycosyl phosphatidyl inositol anchors. AB - A variable concept for the synthesis of branched glycosyl phosphatidyl inositol (GPI) anchors was established. Its efficiency could be shown by the successful synthesis of the GPI anchor of rat brain Thy-1 and of the scrapie prion protein both in the water soluble 1c and lipidated form 1a. Retrosynthesis led to building blocks 2-6 of which 5 could be further disconnected to building blocks 7 9. Trichloroacetimidate 5 was built up in a straightforward manner starting from glycosyl acceptor 9 using known glycosyl donors 7 and 8. The carbohydrate backbone was then assembled by glycosylation of pseudodisaccharide acceptor 6 with donor 5. To ensure high stereoselectivity and good yields in the glycosylation reactions, anchimeric assistance was employed. Successive deprotection and introduction of the various phosphate residues gave the fully protected GPI anchors. Catalytic hydrogenation and acid-catalyzed cleavage of the Boc protecting groups afforded the target molecules, which could be fully structurally assigned. PMID- 12585869 TI - Allylic-type diindium reagents. Reactivity toward electrophiles and cascade coupling reactions with imines. AB - The allylic-type diindium reagents A and B were prepared from 3-bromo-1 iodopropene (1a) and 4-bromo-2-iodobut-2-ene (1b), respectively, and their reactions with electrophiles were investigated. The diindium reagents A and B were initially reacted with imines and the resulting vinylindium compounds were then treated with organic halides in the presence of Pd(PPh(3))(4) to give linear N-aryl and N-tosyl homoallylic amines. Diindium A is stable in a small amount of water in solvent, whereas B is easily protonated to give a crotylindium reagent. The reaction of B with benzaldehyde gives mainly the 1,3- and 1,5-diols via a spontaneous coupling with two molecules of the aldehyde, in contrast to A, which reacts with one molecule of carbonyl compounds to give the vinylindium compounds. PMID- 12585870 TI - Fairly marked enantioselectivity for the hydrolysis of amino acid esters by chemically modified enzymes. AB - The hydrolysis (deacylation) of enantiomeric substrates by the chemically modified enzymes decanoyl-alpha-chymotrypsin and decanoyl-trypsin was studied. Reaction activity for decanoyl-alpha-chymotrypsin was lower than that for the native enzyme, although intriguingly the enantioselectivity was markedly enhanced as compared with the native enzyme. In particular, the apparently complete enantioselective catalysis was attained for the hydrolytic cleavage of p nitrophenyl N-dodecanoyl-D(L)-phenylalaninates. The enhancement of enantioselectivity, however, was not observed for decanoyl-trypsin. These results suggest that the chemically modified alpha-chymotrypsin by addition of hydrophobic groups has promoted enantioselectivity for the hydrolysis of hydrophobic esters. PMID- 12585871 TI - Stereochemistry of the allylation and crotylation reactions of alpha-methyl-beta hydroxy aldehydes with allyl- and crotyltrifluorosilanes. Synthesis of anti,anti dipropionate stereotriads and stereodivergent pathways for the reactions with 2,3 anti- and 2,3-syn-alpha-methyl-beta-hydroxy aldehydes. AB - A new method for the stereoselective synthesis of the anti,anti-dipropionate stereotriad via the reaction of alpha-methyl-beta-hydroxy aldehydes with (Z) crotyltrifluorosilane (24) is described. These reactions were designed to occur through bicyclic transition states (e.g., 31) in which the silane reagent is covalently bound to the beta-hydroxyl group of the aldehyde and the crotyl group is transferred intramolecularly. This methodology was used to synthesize the C(7) C(16) segment (58) of zincophorin, which contains a synthetically challenging all anti stereopentad unit. Surprisingly, 2,3-anti- and 2,3-syn-alpha-methyl-beta hydroxy aldehydes react in a stereodivergent manner with 24: 2,3-anti-beta hydroxy aldehydes give the targeted anti,anti-dipropionate adducts with high selectivity, but the reactions of 2,3-syn-beta-hydroxy aldehydes are poorly selective. The stereodivergent behavior of 2,3-syn- vs 2,3-anti-alpha-methyl-beta hydroxy aldehydes is also exhibited in their reactions with the allyl- (68) and (E)-crotyltrifluorosilanes (27). Competition experiments performed with beta hydroxy aldehydes 37a (anti) and the corresponding p-methoxybenzyl (PMB) ether 48, and between aldehyde 39 (syn) and the PMB ether 90, established that the 2,3 anti-beta-hydroxy aldehydes react predominantly through bicyclic transition states while the 2,3-syn aldehydes react predominantly through conventional Zimmerman-Traxler transition states. NMR studies established that both the 2,3 syn and the 2,3-anti aldehydes form stable, pentavalent silicate intermediates (98 and 100) with PhSiF(3), but chelated structures 99 and 101 could not be detected. The activation energies for the competing bicyclic and conventional Zimmerman-Traxler transition states were calculated by using semiemperical methods (MNDO/d). These calculations indicate that the stereodivergent behavior of the 2,3-syn-beta-hydroxy aldehydes and the 2,3-anti-beta-hydroxy aldehydes is due to differences in nonbonded interactions in the bicyclic transition states. Specifically, nonbonded interactions in the bicyclic transition states for the allylation/crotylation reactions of the 2,3-syn-beta-hydroxy aldehydes permits the traditional Zimmerman-Traxler transition states to be preferentially utilized. PMID- 12585872 TI - Cyclization of aryllithiums tethered to methylenecycloalkanes: stereoselective synthesis of 4a-substituted cis-hexahydrofluorenes. AB - The cyclization of an aryllithium tethered to a methylenecycloalkane, generated from 2-(o-bromobenzyl)-1-methylenecycloalkanes 1, 2, and 3 by low-temperature lithium-bromine exchange, has been found to be a kinetically slow but thermodynamically favorable process that proceeds at a convenient rate in an exclusively 5-exo fashion when solutions of the aryllithium in n-heptane-di-n butyl ether (9:1 v/v) are warmed to 45 degrees C. The cyclization affords stereoisomerically pure cis-fused products (7 and 8) when the methylenecycloalkane is five- or six-membered but it is less stereoselective when the methylenecycloalkane is seven-membered. The ring-closure of the aryllithium derived from 2-(o-bromobenzyl)-1-methylenecyclohexane (2) provides an experimentally convenient route to stereoisomerically pure 4a-substituted cis hexahydrofluorenes in 60-90% isolated yield. PMID- 12585873 TI - Synthesis of unsymmetrically substituted 1,3-butadiynes and 1,3,5-hexatriynes via alkylidene carbenoid rearrangements. AB - Unsymmetrically substituted 1,3-butadiynes and 1,3,5-hexatriynes are synthesized in four steps from commercially available aldehydes or carboxylic acids. The key step in this process involves a Fritsch-Buttenberg-Wiechell rearrangement, in which an alkylidene carbenoid intermediate subsequently rearranges to the desired polyyne. This rearrangement proceeds under mild conditions, and it is tolerant of a range of functionalities. In general, the procedurally facile formation of the dibromoolefinic precursors, in combination with the effectiveness of the rearrangement step, makes this procedure an attractive alternative to traditional methods for di- and triyne synthesis that utilize palladium or copper catalysis. PMID- 12585874 TI - Stereoselective synthesis of conformationally constrained glycosylated amino acids using an enzyme-catalyzed desymmetrization. AB - As part of an effort to probe the mechanism by which glycosyltransferases recognize glycoproteins and assemble the core structures of O-linked oligosaccharides, constrained glycopeptides, compounds 2 and 3, based on the alpha-N-acetylgalactosaminyl serine substructure 1, were designed. In this paper we describe a stereoselective preparation of protected versions of these compounds. A pig liver esterase-catalyzed enzymatic desymmetrization of a diacetate substrate, 10, was employed as a key component in the synthesis. PMID- 12585875 TI - Copper(II)-mediated autoxidation of tert-butylresorcinols. AB - Although copper(II)-mediated oxidation of phenols results in oxidative coupling rather than in oxygenation, it was recently reported that naturally occurring 5 alkylresorcinols undergo oxygenation in the presence of copper(II). To explore the generality of this reaction, the copper(II)-mediated autoxidation of 4-tert butylresorcinol and 4,6-di-tert-butylresorcinol was investigated and was found to result in direct oxygenation at open activated positions and, at the tert-butyl substituted positions, in oxygenation with competing loss of (as isobutylene) and 1,2-rearrangement of the tert-butyl group. 5-tert-Butyl-2-hydroxy-1,4 benzoquinone is the major product from both starting materials, and the final product mixture reflects, in part, coupling of metastable initially formed electrophilic and nucleophilic side products. Mechanisms that are consistent with the observed products and control reactions are proposed. The key step appears to be equilibration of a copper(II)-resorcinolate with a charge-transfer radical form that reacts regioselectively with O(2) as prescribed by resonance. PMID- 12585876 TI - Synthesis of novel hapten derivatives of 1alpha,25-dihydroxy-vitamin D3 and its 20-epi analogue. AB - Hapten derivatives of 1alpha,25-dihydroxyvitamin D(3) and its 20-epimer were synthesized and conjugated to a carrier protein for raising polyclonal antibodies. The haptens were linked through spacers at C-16, thereby exposing both the A-ring and the side chain of the molecules, to maximize antibody specificity. The spacers were introduced via stereoselective hydroboration of 16 ene intermediates as the key step. In immunoassays, the antibodies raised toward the natural hormone were selective to this compound over derivatives with modifications in the A-ring or the side chain. The antibodies toward the 20 epimer, however, were unable to recognize modifications in the side chain. PMID- 12585877 TI - Efficient construction of the bicyclo[5.3.0]decenone skeleton based on the Rh(I) catalyzed allenic Pauson-Khand reaction. AB - A reliable procedure for constructing a bicyclo[5.3.0]deca-1,7-dien-9-one ring system by rhodium-catalyzed Pauson-Khand reaction (PKR) of allenynes with a sulfonyl group has been developed. Investigation of the rhodium-catalyzed PKR on 19 examples of 1,2-nonadien-8-yne derivatives demonstrated that (i) acceptable yields could be consistently achieved through the proper choice of the rhodium catalyst ([RhCl(CO)(2)](2) or [RhCl(CO)dppp](2)) depending on the starting allenyne and that (ii) an ester functionality as well as hydroxy and siloxy groups could be tolerated in this rhodium-catalyzed PKR. PMID- 12585878 TI - A study into the possible homoaromatic nature of some related carbene and cationic intermediates with the potential for transannular interaction. AB - A comparison of the aromatic nature of the cations and carbenes centered at the 8 position of tricyclo[3.2.1.0(2,4)]octane, the 9-position of pentacyclo[4.3.0.0(2,4).0(3,8).0(5,7)]nonane, the 6-position of tricyclo[3.1.1.0(2,4)]heptane, and the 4-position of tetracyclo[3.3.0.0(2,8).0(3,6)]octane has been undertaken. Hybrid density functional theory calculations have been used to examine the geometric, energetic, and magnetic characteristics of each cation, singlet carbene, and triplet carbene. The results shed light on the flexibility of the polycycles to achieve stable intermediates and on the breadth of homoaromaticity. PMID- 12585879 TI - Michael addition of chloroalkyloxazolines to electron-poor alkenes: synthesis of heterosubstituted cyclopropanes. AB - Lithiated 2-(1-chloroethyl)-4,4-dimethyl-2-oxazoline 7 adds to electron-poor alkenyl heterocycles to afford substituted cyclopropanes in excellent yields. A route to chiral nonracemic heterosubstituted cyclopropanes, starting from optically active 2-chloromethyl-2-oxazolines, is highlighted as well. PMID- 12585880 TI - Enantioselective synthesis of spirocyclic aminochroman derivatives according to the CN(R,S) strategy. AB - Enantiomerically pure (3'R)- and (3'S)-3',4'-dihydrospiro[piperidine-2,3'(2'H) benzopyran]s (R)-10 and (S)-10 were successfully synthesized according to the CN(R,S) methodology with the aim of serving as a pattern for the generation of related spirocyclic compounds. Two different synthetic pathways were studied starting from 2-cyano-6-phenyloxazolopiperidine (-)-2. One of them was selected and used for the preparation of amines (R)-17 and (S)-17 starting from (-)-2 and (+)-2, respectively. The enantiomeric purity of all final aminochroman derivatives was determinated by capillary electrophoresis using beta-cyclodextrin as the chiral selector. PMID- 12585881 TI - Functionalized cis- and trans-fused bicyclic alpha-amino acids via stereoselective double annulation and dequaternization reactions. AB - Fused bicyclic alpha-amino acids can be prepared by a double Michael reaction of p-anisyl ethynyl ketone and a tethered diacid, cyclization via hydrogenation or hydration of a CN group, and oxidation of the p-anisyl group. The substitution level of the alpha-amino acids can be adjusted by decyanation or decarboethoxylation of the intermediates. Bicyclic alpha-amino acids prepared in this way include cis- and trans-perhydroisoquinoline-3-carboxylic acids and cis perhydro-2-pyrindine-3-carboxylic acids of various substitutions and oxidation levels. The bicyclic alpha-amino acids may be regarded as functionalized and conformationally restricted analogues of proline, pipecolic acid, 2-aminoadipic acid, or glutamic acid. PMID- 12585882 TI - Concise and stereocontrolled synthesis of pseudo-C2-symmetric diamino alcohols and triamines for use in HIV protease inhibitors. AB - A new protocol is described for the stereocontrolled synthesis of pseudo-C(2) symmetric core units of interest as candidates for HIV protease inhibition. Addition of unbranched and branched organolithium reagents to cyanohydrins from l phenylalaninal and l-isoleucinal, followed by in situ reduction of the intermediate imines and CHT deprotection under MW irradiation, led to 1,3-diamino alcohols 6a and 8a as the major products in satisfactory to good yields. The first preparation of a previously unreported pseudo-C(2)-symmetric triamino derivative was accomplished expeditiously via high-yielding nitro-Mannich addition of the silylnitronate, from 2-phenyl-1-nitroethane, to the PMP imine derived from l-phenylalaninal. Reduction of the nitro group in the moderately unstable nitro diamine adduct, followed by chromatographic separation of the required diastereoisomer and CHT debenzylation under MW irradiation, led to the 2 PMP-protected triamine 19 isolated as a bis(sulfonamide). PMID- 12585884 TI - Highly diastereoselective oxidation of 2-amino-2-deoxy-1-thio-beta-D glucopyranosides: synthesis of imino sulfinylglycosides. AB - A synthetic route to imino thioglycosides and to imino sulfinylglycosides has been developed. A detailed study on the diastereoselective oxidation of 2-amino-2 deoxy-1-thio-beta-D-glucopyranosides is reported. It has been shown that the stereochemical outcome of the oxidation is highly dependent on the protective group of the amine function. While the oxidation of iminothioglycosides is slightly diastereoselective (up to 30% de in favor of the R(S) sulfoxide), a single isomer is obtained in the case of tetrachlorophthalimido derivatives. The absolute configuration of the sulfinyl glycoside was ascertained by NMR analysis using our recent model on the basis of the exo-anomeric effect corroborated by X ray crystallography. PMID- 12585883 TI - Asymmetric synthesis of unusual fused tricyclic beta-lactam structures via aza cycloadditions/ring closing metathesis. AB - Conveniently substituted bis-beta-lactams, pyrrolidinyl-beta-lactams, and piperidinyl-beta-lactams undergo ring-closing methatesis using Grubbs' carbene, Cl(2)(Cy(3)P)(2)Ru=CHPh, to give medium-sized rings fused to bis-2-azetidinone, pyrrolidinyl-2-azetidinone, or piperidinyl-2-azetidinone systems. The diolefinic cyclization precursors can be obtained from optically pure 4-oxoazetidine-2 carbaldehydes bearing an extra alkene tether at position 1 or 3 of the beta lactam ring via [2 + 2] cycloaddition of imino 2-azetidinones, N-metalated azometine ylide [3 + 2] cycloaddition, and subsequent N-acylation of the pyrrolidinyl nitrogen atom, or through aza-Diels-Alder cycloaddition of 2 azetidinone-tethered imines. Under standard reaction conditions, the combination of cycloaddition reactions of 2-azetidinone-tethered imines with ring-closing methatesis offers an asymmetric entry to a variety of unusual fused tricyclic 2 azetidinones bearing two bridgehead nitrogen atoms. PMID- 12585885 TI - Efficient conversion of sulfones into beta-keto sulfones by N-acylbenzotriazoles. AB - Acyclic sulfones 4a-f and alicyclic sulfone 7 react with readily available N acylbenzotriazoles 3a-g (derived from aliphatic, aromatic, and heteroaromatic carboxylic acids) to provide the corresponding beta-keto sulfones 5a-n and 8a-c, respectively, in good to excellent yields. PMID- 12585886 TI - Regioselective photocycloaddition of pyridine derivatives to electron-rich alkenes. AB - Irradiation of a benzene solution of 3-cyano-2,6-dimethoxypyridine in the presence of ethyl vinyl ether (EVE) gave 1:1 photoadducts, 3-cyano-5-ethoxy-2,8 dimethoxy-4,5-dihydroazocine, in good yields, whose structure was established by X-ray single-crystal analysis. The photoadduct was produced via cycloaddition between the C3-C4 position of the pyridine derivatives and an alkene chromophore. On the other hand, 3-cyano-2,6-dimethoxy-4-methylpyridine cycloadds to EVE at the C2-C3 position of the pyridine ring upon irradiation. The difference is explained on the basis of the steric effect. PMID- 12585887 TI - C10-substituted camphors and fenchones by electrophilic treatment of 2 methylenenorbornan-1-ols: enantiospecificity, scope, and limitations. AB - Valuable chiral sources of C(10)-substituted camphors and C(10)-substituted fenchones can be straightforwardly obtained by treatment of an appropriate, easily obtainable, camphor- or fenchone-derived 2-methylenenorbornan-1-ol with an electrophilic reagent. The process takes place via a tandem regioselective carbon carbon double-bond addition/stereocontrolled Wagner-Meerwein rearrangement. A complete study of the enantiospecificity, scope, and limitations of this process, as well as about the role played by the hydroxyl group attached at the C(1) bridgehead position of the starting 2-methylenenorbornan-1-ols, has been realized. The feasibility of the described methodology has been exemplified by the highly efficient enantiospecific preparation of several interesting C(10) halogen-, C(10)-O-, C(10)-S-, C(10)-Se-, or C(10)-C-substituted camphors and fenchones. PMID- 12585888 TI - New mixed phosphonate esters by transesterification of pinacol phosphonates and their use in aldehyde and ketone coupling reactions with nonstabilized phosphonates. AB - Alkylpinacol phosphonates were prepared by rhodium-catalyzed olefin hydrophosphorylation, and attempted alpha-deprotonation of the pinacol derived alkyl phosphonates resulted in ring cleavage. The propensity of the alkylpinacol phosphonates to undergo ring opening was exploited to prepare phosphonic acid monomethyl esters in high yield by transesterification in acidulated methanol. Esterification and alkylation with aldehydes or ketones gave beta-hydroxy mixed phosphonate esters. tert-Butyl and benzylic phosphonate ester protective groups were introduced to improve the efficiency and functional group compatibility of beta-hydroxy phosphonate saponification. The beta-hydroxy phosphonic acid monomethyl esters were dehydrated with diisopropylcarbodiimide, which gave oxaphosphetane intermediates that collapse to an olefin. The overall reaction sequence complements the arsenal of Horner-Wadsworth-Emmons-type coupling reactions. PMID- 12585889 TI - Spin densities in dialkoxy-[16]annulene anion radicals: dimerization of alkoxy [8]annulenes. AB - The anion radicals of alkoxy-substituted cyclooctatetraenes in hexamethylphosphoramide spontaneously dimerize to form the dianions of dialkoxy [16]annulenes. The dianions reveal the expected high-field NMR resonance for the internal protons. After electron transfer, the EPR spectra of the corresponding anion radicals reveal that only the 1,5-dialkoxy systems are formed. Further, the measured proton and (13)C spin densities show that the odd electron resides in a molecular orbital with six hydrogens in "deep" nodal positions that completely hide them from EPR detection. This MO corresponds to the nonbonding (singly occupied) MO of higher energy after splitting of the degenerate nonbonding MOs by the two-electron-withdrawing substituents. The surprising electron-withdrawing nature of the alkoxy substituents is attributed to a rather strong mixing of the sigma and pi systems in [16]annulene. PMID- 12585890 TI - 2-Quinolyl- and 1-isoquinolylnitrenes: ring expansion and ring opening in heteroarylnitrenes. AB - Argon matrix photolysis of tetrazolo[1,5-a]quinoline 8 and tetrazolo[5,1 a]isoquinoline 7 causes nitrogen elimination and ring expansion to 1,3 diazabenzo[d]cyclohepta-1,2,4,6-tetraene 13. The photolysis of tetrazolo[5,1 a]isoquinoline 7 also causes ring opening to o-cyanophenylketenimine 22. Mechanisms of ring opening of heteroarylnitrenes are discussed. PMID- 12585891 TI - Are neutral oxocarbons stable? AB - A set of novel oligomeric polycyclic neutral oxocarbons has been computed at the B3LYP level of density functional theory. On the basis of the calculated dissociation enthalpies, the most stable structures are C(9)O(9) and C(12)O(12) with benzene-like central ring, in agreement with the experimental finding, and the enhanced stability is due to their aromaticity. Several other oxocarbons also have been identified as stable and are targets for further experimental investigations. PMID- 12585892 TI - Directed ring-opening of 1,5-dioxaspiro[3.2]hexanes: selective formation of 2,2 disubstituted oxetanes. AB - 1,5-Dioxaspiro[3.2]hexanes undergo ring-opening reactions with many heteroatom nucleophiles to provide alpha-substituted-beta'-hydroxy ketones. However, certain Lewis acidic nucleophiles provide 2,2-disubstituted oxetanes. Herein, the results of reactions of 3-phenyl-1,5-dioxaspiro[3.2]hexane with a variety of nitrogen containing heteroaromatic bases are reported. There appears to be a correlation between the pK(a) of the nucleophile and the reaction outcome with more acidic nucleophiles providing 2,2-disubstituted oxetanes. Moreover, the mode of ring opening can be directed toward the substituted oxetane by the addition of a Lewis acid. These results are rationalized by calculation of stationary points on the potential energy surfaces for the various possible reaction pathways using ab initio molecular orbital methods. PMID- 12585893 TI - Rational design, synthesis, and pharmacological evaluation of 2-azanorbornane-3 exo,5-endo-dicarboxylic acid: a novel conformationally restricted glutamic acid analogue. AB - The design and synthesis of conformationally restricted analogues of alpha-amino acids is an often used strategy in medicinal chemistry research. Here we present the rational design, synthesis, and pharmacological evaluation of 2-azanorbornane 3-exo,5-endo-dicarboxylic acid (1), a novel conformationally restricted (S) glutamic acid (Glu) analogue intended as a mimic of the folded Glu conformation. The synthesis of 1 was completed in its racemic form in eight steps from commercially available starting materials. As a key step, the first facially selective hydroboration of a 5-methylidene[2.2.1]bicyclic intermediate was investigated. In this transformation, the catalytic methodology of Wilkinson's/catechol borane proved superior to stoichiometric borane or dialkyl borane reagents, in terms of higher diastereomeric excess and chemical yield. To our surprise (+/-)-1 did not show affinity in binding studies on native 2-amino-3 (3-hydroxy-5-methyl-4-isoxazolyl)propionic acid (AMPA) (IC(50) > 300 microM, [(3)H]AMPA) or kainic acid (IC(50) > 160 microM, [(3)H]kainic acid) receptors nor in binding studies on the cloned iGluR5,6 subtypes (IC(50) > 300 microM, [(3)H]kainic acid). PMID- 12585894 TI - Computational study of the aminolysis of esters. The reaction of methylformate with ammonia. AB - The aminolysis of esters is a basic organic reaction considered as a model for the interaction of carbonyl group with nucleophiles. In the present computational study the different possible mechanistic pathways of the reaction are reinvestigated by applying higher level electronic structure theory, examining the general base catalysis by the nucleophile, and a more comprehensive study the solvent effect. Both the ab initio QCISD/6-31(d,p) method and density functional theory at the B3LYP/6-31G(d) level were employed to calculate the reaction pathways for the simplest model aminolysis reaction between methylformate and ammonia. Solvent effects were assessed by the PCM method. The results show that in the case of noncatalyzed aminolysis the addition/elimination stepwise mechanism involving two transition states and the concerted mechanism have very similar activation energies. However, in the case of catalyzed aminolysis by a second ammonia molecule the stepwise mechanism has a distinctly lower activation energy. All transition states in the catalyzed aminolysis are 10-17 kcal/mol lower than those for the uncatalyzed process. PMID- 12585895 TI - Facile one-pot coupling-aminovinylation approach to push-pull chromophores: alkyne activation by sonogashira coupling. AB - A straightforward coupling-aminovinylation sequence of terminal alkynes 1, electron-deficient heteroaryl halides 2, and secondary amines 4 furnishes highly solvochromic push-pull chromophores 5 in good yields. Semiempirical calculations (PM3) suggest that the aminovinylation proceeds in a stepwise fashion through a zwitterionic intermediate with a final rate-determining intramolecular protonation. Crucial parameters for the success of the amine addition are the relative LUMO energies and the charge distribution at the beta-alkynyl carbon atom. PMID- 12585896 TI - Rigid-core fluorescent oligothiophene-S,S-dioxide isothiocyanates. Synthesis, optical characterization, and conjugation to monoclonal antibodies. AB - In this paper we report the synthesis of a new class of fluorescent thiophene based isothiocyanates containing a 3,5-disubstituted dithieno[3,2-b:2',3' d]thiophene-4,4-dioxide moiety as the rigid core, using the palladium-catalyzed cross-coupling reaction of aryl stannanes with aryl bromides (Stille coupling). By changing the molecular structure through the progressive addition of thienylene or phenylene units, light emission from blue to orange was obtained. Photoluminescence quantum yields ranged from 0.65 to 0.90 for blue and green light emitters to 0.10-0.35 for yellow and orange ones. Optically and chemically stable fluorescent bioconjugates were prepared by spontaneous reaction of the isothiocyanates with monoclonal antibodies anti-CD3 and anti-CD8 in slightly basic solutions. PMID- 12585897 TI - Total synthesis of (s)-(+)-citreofuran by ring closing alkyne metathesis. AB - A concise total synthesis of citreofuran 4 is described, a structurally unique octaketide derivative belonging to the curvularin family. Key steps involve the elaboration of orsellinic acid methyl ester 5 to acid 14, which converts, on attempted formation of the corresponding acid chloride, to the 3 alkoxyisocoumarin derivative 20. This heterocycle can be used as an activated ester to give ketone 21 on treatment with 3-pentynylmagnesium bromide in the presence of TMSCl as the activating agent. Ring- closing alkyne metathesis (RCAM) of diyne 21 catalyzed by (tBuO)(3)W[triple bond]CCMe(3) affords the strained cycloalkyne 22. Treatment with acid renders its triple bond susceptible to nucleophilic attack by the adjacent carbonyl group, thus leading to a transannular cycloaromatization with formation of the intact skeleton of citreofuran. An X-ray crystallographic study reveals conformational details about this natural product. Finally, it is shown that 4 as well as its protected precursor 23 are able to cleave double-stranded DNA under oxidative conditions. PMID- 12585898 TI - A repertoire of pyridinium-phenyl-methyl cross-talk through a cascade of intramolecular electrostatic interactions. AB - Direct intramolecular cation-pi interaction between phenyl and pyridinium moieties in 1a(+) has been experimentally evidenced through pH-dependent (1)H NMR titration. The basicity of the pyridinyl group (pK(a) 2.9) in 1a can be measured both from the pH-dependent chemical shifts of the pyridinyl protons as well as from the protons of the neighboring phenyl and methyl groups as a result of electrostatic interaction between the phenyl and the pyridinium ion in 1a(+) at the ground state. The net result of this nearest neighbor electrostatic interaction is that the pyridinium moiety in 1a becomes more basic (pK(a) 2.92) compared to that in the standard 2a (pK(a) 2.56) as a consequence of edge-to-face cation (pyridinium)-pi (phenyl) interaction, giving a free energy of stabilization (DeltaDeltaG(o)pKa) of -2.1 kJ mol(-1). The fact that the pH dependent downfield shifts of the phenyl and methyl protons give the pK(a) of the pyridine moiety of 1a also suggests that the nearest neighbor cation (pyridinium) pi (phenyl) interaction also steers the CH (methyl)-pi (phenyl) interaction in tandem. This means that the whole pyridine-phenyl-methyl system in 1a(+) is electronically coupled at the ground state, cross-modulating the physicochemical property of the next neighbor by using the electrostatics as the engine, and the origin of this electrostatics is a far away point in the molecule-the pyridinyl nitrogen. The relative chemical shift changes and the pK(a) differences show that the cation (pyridinium)-pi (phenyl) interaction is indeed more stable (DeltaDeltaG(o)pKa = -2.1 kJ mol(-1)) than that of the CH (methyl)-pi (phenyl) interaction (DeltaDeltaG(o)pKa = -0.8 kJ mol(-1)). Since the pK(a) of the pyridine moiety in 1a is also obtained through the pH-dependent shifts of both phenyl and methyl protons, it suggests that the net electrostatic mediated charge transfer from the phenyl to the pyridinium and its effect on the CH (methyl)-pi (phenyl) interaction corresponds to DeltaG(o)pKa of the pyridinium ion (approximately 17.5 kJ mol(-1)), which means that the aromatic characters of the phenyl and the pyridinium rings in 1a(+) have been cross-modulated owing to the edge-to-face interaction proportional to this DeltaG(o)pKa change. PMID- 12585899 TI - Computational studies on pseudorotaxanes by molecular dynamics and free energy perturbation simulations. AB - A computational scheme that comprises the utilization of the AMBER force field with RESP charges and an explicit solvent model for acetonitrile proved to be useful for studying the structures and energetics of pseudorotaxanes of benzidine and 4,4'-biphenol with cyclobis(paraquat-p-phenylene). The scheme can be further utilized for modeling [2]rotaxanes. PMID- 12585900 TI - Cyanomethyl anion/carbon dioxide system: an electrogenerated carboxylating reagent. Synthesis of carbamates under mild and safe conditions. AB - A new carboxylating reagent ((-)CH(2)CN/CO(2)) was obtained by bubbling CO(2) in a CH(3)CN-TEAP (tetraethylammonium perchlorate) solution previously electrolyzed under galvanostatic control. Organic carbamates were isolated from these solutions after addition of amines and an alkylating agent. In this paper, we describe the optimized conditions for the electrochemical synthesis of carbamates from amines and CO(2), in mild and safe conditions, without any addition of bases, probases, or catalysts. Carbamates were isolated from primary and secondary aliphatic amines in high to excellent yields and from aromatic amines in moderate yields (dependent on the nucleophilicity of the nitrogen atom). PMID- 12585902 TI - Deoxygenation of dithiirane 1-oxides with Lawesson's reagent leading to the corresponding dithiiranes. AB - 3,3-Disubstituted dithiirane 1-oxides were efficiently reduced with Lawesson's reagent (LR) to give the corresponding dithiiranes. X-ray diffraction analysis of 3,3-di(1-adamantyl)dithiirane is reported. Reaction of (34)S-labeled 3,3-di(1 adamantyl)dithiirane 1-oxide with LR produced the corresponding dithiirane in which the (34)S atoms were retained quantitatively. PMID- 12585901 TI - Application of peptidyl radicals into a new radical cascade leading to unsaturated gamma-lactams. AB - Radical cyclization of dipeptides 1a-h proceeds smoothly to give five- and seven membered rings in good to moderate total yields using Stork's catalytic tin hydride method. A radical is generated on a protecting group and translocated to the peptide moiety. Following a cyclization reaction, the vinyl radical can abstract hydrogen from a benzyl group on an amine, which results in elimination of the protected amine group. Encouraging results have notably been obtained with amino acids other than glycine. PMID- 12585903 TI - Chemical fixation of carbon dioxide catalyzed by binaphthyldiamino Zn, Cu, and Co salen-type complexes. AB - Binaphthyldiamino salen-type Zn, Cu, and Co complexes can efficiently catalyze reactions of epoxides with carbon dioxide in the presence of various catalytic amounts of organic bases. The simplest binaphthyldiamino salen-type Zn complex gave the five-membered cyclic carbonate 2 in excellent yield in the presence of triethylamine. A Lewis acid and Lewis base cocatalyzed mechanism is proposed. PMID- 12585904 TI - Use of a boroxazolidone complex of 3-iodo-L-tyrosine for palladium-catalyzed cross-coupling. AB - Complexation of 3-iodo-L-tyrosine with 9-borabicyclo[3.3.1]nonane (9-BBN) provides a convenient substrate for a palladium-catalyzed coupling reaction. The complex is stable to silica gel chromatography (hexanes/ethyl acetate), dilute triethylamine in THF, and potassium fluoride in DMF. The desired product, 3 ethynyl-L-tyrosine, was released from the complex by simply diluting its solution in methanol with chloroform. Interestingly, the complex remains stable in solutions of either methanol or chloroform individually. PMID- 12585905 TI - Unusual reaction of chloramine-T with araldoximes. AB - Reaction of araldoximes with 4 equiv of chloramine-T in refluxing methanol produces N-(p-tolyl)-N-(p-tosyl)benzamides via addition of 2 equiv of chloramine T to the intermediate nitrile oxide followed by extrusion of sulfur dioxide. PMID- 12585906 TI - Pd(OH)2/C (Pearlman's catalyst): a highly active catalyst for Fukuyama, Sonogashira, and Suzuki coupling reactions. AB - Treatment of thiol esters 1 with zinc reagent 2 in the presence of a small amount ( 500 nM) induces integrin alphaIIbbeta3 activation and irreversible platelet adhesion independent of PKC. Our studies demonstrate the existence of a complex signalling relationship operating between PKC, cytosolic calcium and integrin alphaIIbbeta3 that serves to regulate platelet-adhesion dynamics under flow. Furthermore, we have established the existence of PKC-dependent and -independent pathways regulating integrin alphaIIbbeta3 activation and stable platelet adhesion on vWf. PMID- 12585967 TI - Performance of health-status scales when used selectively or within multi-scale questionnaire. AB - BACKGROUND: Little work has been done to investigate the suggestion that the use of selected scales from a multi-scale health-status questionnaire would compromise reliability and validity. The aim of this study was to compare the performance of three scales selected from the SF-36 generic health questionnaire when administered in isolation or within the entire SF-36 to patients with musculoskeletal disorders. METHODS: Two groups of patients referred to an orthopedic department completed a mailed questionnaire within 4 weeks prior to and a second questionnaire during their visit. The first group completed three SF 36 scales related to physical health (physical functioning, bodily pain, and general health perceptions) on one occasion and all eight SF-36 scales on the other occasion. The second group completed the entire SF-36 on two occasions. Results for patients who reported unchanged health status and had complete scores were analyzed; 80 patients in the first and 62 patients in the second group. RESULTS: The Cronbach alpha reliability and intraclass correlation coefficients exceeded 0.7 for all three scales for both groups. For the first group the mean difference between the scores was 0.4 point for physical functioning, 2.5 points for bodily pain, and 0.5 point for general health perceptions, which did not differ significantly from the corresponding differences for the second group (0.1, 1.9 and 1 point, respectively). CONCLUSION: The use of selected scales from a multi-scale health-status questionnaire seems to yield similar results compared to their use within the entire questionnaire. PMID- 12585969 TI - Poor response to tuberculosis treatment with regimens without rifampicin in immunosuppressed AIDS patients. AB - A prospective study was conducted on 79 advanced immunosuppressed AIDS patients from 1997 to 1999, during which nine cases of tuberculosis (TB) were diagnosed. The main clinical and laboratory characteristics and the response to TB treatment were reviewed. The clinical manifestations of TB were: pulmonary (six cases), extrapulmonary (two cases) and disseminated (one case). These patients were being treated with highly active antiretroviral treatment (HAART) and were not responding. In three cases an optional regimen without rifampicin (RMP) was indicated to maintain HAART during TB treatment. A clinical response to TB treatment (disappearance of fever) was observed in 6/9 patients during a mean of 73 days (SD = 96). The three unresponsive patients were those treated without RMP. A switch to TB regimens containing RMP was proposed and successful. In our study, though it was limited by a small sample size, the response to TB regimens without rifampin was poor in immunosupressed patients failing HAART. PMID- 12585970 TI - A randomized, pilot trial comparing full versus escalating dose regimens for the desensitization of AIDS patients allergic to sulfonamides. AB - Sulfonamides are drugs extensively used in the management of AIDS patients. However, the use of sulfonamides is often associated with the development of allergic reactions, provoking the substitution of the drug (by another that may be less effective); alternatively attempts are made to desensitize the patient. OBJECTIVE: Compare two drug regimens (full vs. escalating doses) for the oral desensitization of AIDS patients allergic to sulfonamides. MATERIAL AND METHODS: AIDS patients with previous allergic reactions to sulfonamides and requiring prophylaxis against Pneumocistis carinii, central nervous system toxoplasmosis and diarrhea caused by Isospora belli were randomly assigned to a group receiving a routine dose of cothrimoxazole, or another that received escalating doses of an oral suspension of the same drug, initiating with 75 mg/day of sulfamethoxazole that was doubled every 48 hours till the full dose was reached, if no allergic reaction occurred. Patients were monitored for at least 6 months after enrollment in the trial. The major end-point was the ability to maintain prophylactic treatment after that period of time. Plasma viral load (PVL) and CD(4)/CD(8) counts were measured at baseline. Liver enzymes and hematological parameters were measured at baseline and after 1, 3 and 6 months. RESULTS: Eighteen patients were enrolled in the study (15 men and 3 women), with ages ranging from 30 to 57 years (mean 39.9). The mean CD(4) counts were slightly higher for patients receiving a full dose; there was also a trend towards higher baseline CD(8) counts among patients developing new reactions. The mean PVL was similar among the patients in both desensitization groups. The incidence of new allergic reactions was identical (40%) in the two groups. All adverse reactions were mild and no significant increase in liver enzymes were observed. CONCLUSION: Dose regimen is not a predictor of the development of new allergic reactions amongst patients challenged with sulfonamides after an initial allergic reaction. PMID- 12585968 TI - Brn3c null mutant mice show long-term, incomplete retention of some afferent inner ear innervation. AB - BACKGROUND: Ears of Brn3c null mutants develop immature hair cells, identifiable only by certain molecular markers, and undergo apoptosis in neonates. This partial development of hair cells could lead to enough neurotrophin expression to sustain sensory neurons through embryonic development. We have therefore investigated in these mutants the patterns of innervation and of expression of known neurotrophins. RESULTS: At birth there is a limited expression of BDNF and NT-3 in the mutant sensory epithelia and DiI tracing shows no specific reduction of afferents or efferents that resembles neurotrophin null mutations. At postnatal day 7/8 (P7/8), innervation is severely reduced both qualitatively and quantitatively. 1% of myosin VIIa-positive immature hair cells are present in the mutant cochlea, concentrated in the base. Around 20% of immature hair cells exist in the mutant vestibular sensory epithelia. Despite more severe loss of hair cells (1% compared to 20%), the cochlea retains many more sensory neurons (46% compared to 15%) than vestibular epithelia. Even 6 months old mutant mice have some fibers to all vestibular sensory epithelia and many more to the cochlear apex which lacks MyoVIIa positive hair cells. Topologically organized central cochlea projections exist at least until P8, suggesting that functional hair cells are not required to establish such projections. CONCLUSION: The limited expression of neurotrophins in the cochlea of Brn3c null mice suffices to support many sensory neurons, particularly in the cochlea, until birth. The molecular nature of the long term survival of apical spiral neurons remains unclear. PMID- 12585971 TI - Sensitivity pattern of Salmonella serotypes in Northern India. AB - BACKGROUND: Enteric fever continues to be a major public health problem, especially in the developing countries of the tropics. We determined the incidence of Salmonella bloodstream infections and their antimicrobial resistance patterns from May to August in the years 1997-2001 in Haryana, a large state of India. The minimum inhibitory concentration (MIC) was also determined for 60 isolates of S. typhi to various commonly used antimicrobial agents. MATERIAL AND METHODS: Blood cultures of 6,956 patients (PUO/septicemia) were processed by standard procedures and the Salmonella spp. isolates were identified with specific antisera and with standard biochemical tests. Antimicrobial susceptibilities were determined by Stokes disc diffusion method. The MIC of 60 randomly isolated strains of S. typhi was determined by agar dilution method using Mueller Hinton Agar medium. RESULTS: Isolation rates of Salmonella spp. increased in 2000 and 2001. Multidrug resistance (MDR) in S. typhi had increased while in S. paratyphi it had decreased markedly. Ninety per cent chloramphenicol sensitivity was seen in S. typhi by MIC method. There was a decrease in the susceptibility to ciprofloxacin of S. typhi with MIC showing an upward trend. All S. typhi tested were sensitive to third generation cephalosporins and aminoglycosides with MIC well below the breakpoint. DISCUSSION: Our study indicates that MDR in S. typhi is on the rise in our area. There is also re emergence of chloramphenicol sensitivity. Rising MIC values of ciprofloxacin may lead to prolonged treatment, delayed recovery or pose treatment failure. Thus, sensitivity pattern of causative organism must be sought before instituting appropriate therapy to prevent further emergence of drug resistance. PMID- 12585972 TI - Staphylococcus aureus bacteremia: comparison of two periods and a predictive model of mortality. AB - Staphylococcus aureus is an important pathogen causing bacteremia, primarily affecting hospitalized patients. We studied the epidemiology of S. aureus bacteremia, comparing two periods (early and mid 1990s) and developed a predictive model of mortality. A nested case-control was done. All 251 patients over 14 years old with positive blood cultures for S. aureus were selected. MRSA (methicillin resistant S. aureus) was isolated in 63% of the cases. When comparing the two periods MRSA community-acquired bacteremia increased from 4% to 16% (p=0.01). There was no significant difference in the mortality rate between the two periods (39% and 33%, p=0.40). Intravascular catheters provoked 24% of the cases of bacteremia and were associated with the lowest rate of mortality. In a logistic regression analysis, three variables were associated with death: septic shock, source of bacteraemia and resistance to methicillin. The probability of dying among patients with MRSA and those with methicillin sensitive S. aureus bacteraemia ranged from 10% to 90% and from 4% to 76%, respectively, depending on the source of the bacteraemia and the occurrence of septic shock. The MRSA found in Brazil may be a particularly virulent strain. PMID- 12585973 TI - Estimating the length of the first antiretroviral therapy regiment durability in Sao Paulo, Brazil. AB - Brazil was the first country to provide unrestricted, cost-free access to antiretroviral (ARV) medicine for AIDS treatment. However, there is little data about the benefits of such a policy for these patients. We evaluated the duration of benefit obtained with the introduction of ARVs, defined as the durability of the first ARV regiment. We reviewed the medical charts of patients attended from 1996-2000, at the outpatient clinics of the Federal University of Sao Paulo, Brazil. A total of 120 drug-naive HIV-1 infected patients were eligible to participate in the study. About half of the individuals (53%) presented with disease symptoms; 59% of them had CD(4) count below 200 cells/mm(3). Mean estimated duration of the benefit of therapy was 14.1 months. The most used regimen in this cohort was Zidovudine/3TC/Indinavir (26%), followed by Zidovudine/DDI (17%), and Zidovudine/3TC/Nelfinavir (13%). The most frequent cause of interruption of therapy was gastrointestinal intolerance. Use of treatment regimens with three drugs was more effective than with two drugs, but only for patients with CD(4)<200 cells/mm(3) or CV>100,000 copies RNA/mL. However, the use of triple therapy was associated with a significantly higher probability of reaching maximum viral suppression, during a longer period (p<0.05). The patients enrolled in the study benefitted from therapy for a limited time, after the introduction of double or triple antiretroviral therapy. The incidence of adverse events was significantly associated with loss of the benefits provided by the initial therapeutic regimen. PMID- 12585974 TI - Herpes simplex type 2 pneumonia. AB - Extensive reviews of pulmonary infections in AIDS have reported few herpetic infections. Generally these infections are due to Herpes simplex type 1. Pneumonia due to herpes type 2 is extremely rare. We describe a 40 year-old HIV positive woman who complained of fever, cough and dyspnea for seven years. She had signs of heart failure and the appearance of her genital vesicles was highly suggestive of genital herpes. Echocardiography showed marked pulmonary hypertension, right ventricular hypertrophy and tricuspid insufficiency. After a few days of hospitalization she was treated with Aciclovir and later with Ganciclovir. An open pulmonary biopsy revealed an interstitial inflammation, localized in the alveolar walls. Some pulmonary arteries had widened walls and focal hyaline degeneration. Immunohistochemistry indicated that the nuclei had herpes simplex virus type 2 in many endothelial cells (including vessels with widened walls), macrophages in the alveolar septa and pneumocytes. There was clinical improvement after treatment for herpes. We concluded that as a consequence of herpes infection, endothelial involvement and interstitial inflammation supervene, with thickening of vascular walls and partial obliteration of the vessel lumen. A direct consequence of these changes in pulmonary vasculature was pulmonary hypertension followed by heart failure. PMID- 12585975 TI - Severe low back pain due to retroperitoneal abscess formation in diabetic patients. AB - We report two cases of diabetic patients with severe low back pain associated with retroperitoneal abscesses. In the first case, multiple retroperitoneal and spinal epidural abscesses were detected. Paraplegia due to the spinal epidural abscess was not relieved by drainage of the abscess and subsequent antibiotic therapy. In the second case, drainage of the retroperitoneal abscess and antibiotics were immediately started, resulting in successful recovery. Thus, we suggest that if a diabetic patient complains of low back pain, potential abscess formations should be considered and given appropriate treatment before administering epidural anesthetic injections for pain relief. PMID- 12585976 TI - Post-kala-azar dermal leishmaniasis associated with AIDS. AB - Post-kala-azar dermal leishmaniasis (PKDL) is rarely reported in South America. In spite of the fact that there are many reports about the association of visceral leishmaniasis and AIDS, PKDL is very uncommon in HIV-positive patients, and so far only four cases have been documented in the literature. We present another case with unusual clinicopathological aspects. The patient, a 28-year-old male, from Salvador, Bahia (an endemic area) presented with clinical manifestations of visceral leishmaniasis three years after the diagnosis of AIDS. During treatment for visceral leishmaniasis he developed disseminated miliary papules. Microscopically, the skin biopsy showed a "saw-tooth" pattern with a lichenoid mononuclear infiltrate simulating lichen planus. The histopathological diagnosis was achieved through the finding of amastigotes. The authors discuss the clinicopathological aspects of this case based on a review of the specific literature. PMID- 12585977 TI - Reactivation of Chagas' disease leading to the diagnosis of acquired immunodeficiency syndrome. AB - Reactivation of chronic Chagas' disease is a rare condition and occurs only in immunosuppressed patients. We report a case of a patient with a rapid and fatal reactivation of Chagas' disease, manifested by meningoencephalitis, which lead to a diagnosis of acquired immunodeficiency syndrome (AIDS). We believe there is sufficient evidence to include the reactivation of Chagas' disease among the diagnostic criteria of AIDS in Human Immunodeficiency Virus (HIV) infection. PMID- 12585978 TI - Hepatitis in dengue shock syndrome. AB - Dengue fever is the most frequent arbovirus disease in the world and the most important one in terms of morbidity and mortality. Atypical manifestations of dengue have become commonplace during the last few years, including hepatic damage, which manifests mainly by pain in the right hypochondrium and an increase in the levels of aminotransferases. We describe a case of acute hepatitis in a patient with Dengue Shock Syndrome Grade III. We analyzed the clinical and laboratory aspects of this atypical complication of dengue as well as the differential diagnoses. PMID- 12585979 TI - Vaccine prevents genital herpes in subgroup of women. AB - The herpes simplex virus (HSV) type-2 vaccine studied here prevented genital herpes only in women who were seronegative for HSV-1 and HSV-2 at baseline. Ten of these women would need to be vaccinated to prevent 1 case of genital herpes. The vaccine did not prevent infection with HSV-2 in these women. It did not prevent genital herpes in women with other HSV serologic status or in men. The usefulness of this vaccine is limited by the small subgroup in which it is efficacious. Determining which women fall into this subgroup could prove costly. It is possible that asymptomatic infected persons may spread HSV more readily. Emphasis on the use of condoms and antiviral agents should still be the first line in preventing the spread of genital herpes. PMID- 12585980 TI - Detriments of tPA for acute stroke in routine clinical practice. AB - Under optimal conditions, tissue plasminogen activator (tPA) may be a viable option for treatment of acute ischemic stroke; however, this study showed that protocol is not adhered to in practice and that these protocol deviations are associated with increased mortality and other adverse events. Based on these findings, tPA should not be used in routine clinical practice to treat acute stroke until individual hospitals develop protocols to guarantee the medication's appropriate use. PMID- 12585981 TI - Comparing celecoxib with traditional nonsteroidal anti-inflammatory drugs. AB - Celecoxib is as effective as other nonsteroidal anti-inflammatory drugs (NSAIDs) for treating the symptoms of osteoarthritis or rheumatoid arthritis. However, patients taking celecoxib are less likely to discontinue the medication because of gastrointestinal upset than patients taking traditional NSAIDs. Nevertheless, celecoxib does not decrease the incidence of serious gastrointestinal adverse events with long-term therapy. PMID- 12585982 TI - Screening for and treating asymptomiatic bacteriuria not useful in women with diabetes. AB - Women with diabetes mellitus should not be screened or treated for asymptomatic bacteriuria. Unlike other clinical conditions in which screening for asymptomatic urinary tract infection (UTI) has proved valuable (pregnancy, urologic surgery, renal transplantation), women with diabetes derive no meaningful benefit. Previous recommendations by the US Preventive Services Task Force neither recommended for or against screening or treatment of asymptomatic bacteriuria in diabetic women. PMID- 12585983 TI - Negative ELISA D-dimer assay can miss pulmonary embolism. AB - This evaluation of the use of enzyme-linked immunosorbent assay (ELISA) D-dimer test in routine clinical practice supports other evidence that the assay has a high sensitivity to exclude pulmonary embolism in patient populations in which there is clinical suspicion. Nevertheless, the assay incorrectly excluded the diagnosis of pulmonary embolism in 2 cases. Other examples of clinical decision making exist for which the acceptable negative predictive value for screening is set at 100%--eg, the diagnosis of phenylketonuria in newborns. Physicians who do not want to miss cases of acute pulmonary embolism when they clinically suspect the diagnosis should not rely solely on negative D-dimer assay results when the value to rule out the diagnosis is set at 500 ng/mL. If a lower value is used to define normal--eg, 250 ng/mL, as used in other studies-no cases of acute pulmonary embolism would have been missed in this group of patients. Regardless of the cutoff used, the assay will yield many false-positive results. PMID- 12585984 TI - Self-examination does not reduce breast cancer mortality. AB - Breast self-examination does not decrease breast cancer mortality, according to the results of this randomized controlled trial of 266,000 women who were given intensive instruction in breast self-examination. These findings correspond with the US Preventive Services Task Force policy not to recommend breast self examination for the reduction of breast cancer mortality. PMID- 12585985 TI - Digoxin increases mortality among women with congestive heart failure. AB - Digoxin increases mortality in women with congestive heart failure, compared with men; however, the clinical significance of this is unknown since gender is a nonmodifiable risk factor. More importantly, there is a suggestion of harm when looking at women treated with digoxin versus placebo. Since there are other therapies with definite benefit in congestive heart failure (angiotensin converting enzyme inhibitors, beta-blockers, spironolactone), it is prudent to reconsider the use of digoxin in women with ejection fractions less than 45%. PMID- 12585986 TI - Duct tape removes warts. AB - Duct tape (or any durable, occlusive, tacky tape) appears to be at least as effective as traditional cryotherapy for removal of the common wart. It is an unusual and welcome event in health care when a common ailment is proven equally amenable to an inexpensive, tolerable, and safe alternative therapy. PMID- 12585987 TI - HRT and vitamins C and E do not improve coronary disease in women. AB - Hormone replacement therapy (HRT) and antioxidant vitamin supplements (vitamins E and C) do not provide cardiovascular benefit for postmenopausal women with known coronary heart disease. Moreover, a potential for harm exists with each of the treatments. Therefore, neither should be prescribed specifically for cardiovascular benefit for postmenopausal women with coronary heart disease. PMID- 12585988 TI - Densitometry identifies women in whom treatment will reduce fracture risk. AB - Despite lack of research on the effectiveness of osteoporosis screening to reduce fractures, there is sufficient evidence that bone density measurements accurately predict short-term fracture risk and that treating asymptomatic women with osteoporosis reduces fracture risk. According to this report, a reasonable recommendation is to screen all women older than 65 years and postmenopausal women younger than 65 years who have low weight (or body mass index) or who have never used hormone replacement therapy. The US Preventive Services Task Force noted that the optimal screening frequency has not been studied, but suggested a frequency of not more than every 2 years for older women or every 5 years for younger postmenopausal women. Also of note: other sources, notably the bisphosphonates package labeling, advise against monitoring therapy with repeated dual-energy x-ray absorptiometry or other methods. PMID- 12585990 TI - A thorough yet efficient exam identifies most problems in school athletes. AB - A complete medical history, preferably from the student and a parent, will reveal approximately 75% of problems affecting initial athletic participation. For asymptomatic athletes with no previous injuries, a 90-second screening musculoskeletal test will detect 90% of significant musculoskeletal injuries. A routine screening need not include noninvasive cardiac testing or laboratory tests such as urinalysis, blood count, chemistry profile, lipid profile, ferritin level, or spirometry. PMID- 12585989 TI - Depression screening: a practical strategy. AB - A 2-stage strategy, combining an assessment of severity with depression criteria, can help a physician focus on the most severe cases without missing less severe ones that still need treatment. Because of its brevity, relatively high positive predictive value, and ability to inform the clinician on both depression severity and diagnostic criteria, the PRIME-MD Patient Health Questionnaire (PHQ-9) is the best available depression screening tool for primary care. One-time screening is cost-effective; physicians may elect to screen more often based on risk factors. PMID- 12585991 TI - Abnormal fingernails. PMID- 12585992 TI - Why do physicians think parents expect antibiotics? What parents report vs what physicians believe. AB - OBJECTIVE: To examine the relation between parent expectations for antibiotics, parent communication behaviors, and physicians' perceptions of parent expectations for antibiotics. STUDY DESIGN: A nested cross-sectional study with parallel measures of parents presenting children for acute respiratory infections (previsit) and physicians (postvisit) and audiotaping of the encounters. POPULATION: Ten physicians in 2 private pediatric practices (1 community-based and 1 university-based) and a consecutive sample of 306 eligible parents (response rate, 86%) who were attending sick visits for their children between October 1996 and March 1997. OUTCOMES MEASURED: Communication behaviors used by parents expecting antibiotics and physicians' perceptions of parents' expectations. RESULTS: Parents' use of "candidate diagnoses" during problem presentation increased the likelihood that physicians would perceive parents as expecting antibiotics (from 29% to 47%; P=.04), as did parents' use of "resistance to the diagnosis" (an increase from 7% to 20%). In the multivariate model, parents' use of candidate diagnoses increased the odds that a doctor would perceive a parental expectation for antibiotics by more than 5 times (odds ratio, 5.23; 95% confidence interval, 3.74-7.31; Plt.001), and parents' use of resistance to a viral diagnosis increased these odds by nearly 3 times (odds ratio, 2.73; 95% confidence interval, 1.97-3.79; Plt.001). CONCLUSIONS: Parents perceived as expecting antibiotics may be seeking reassurance that their child is not seriously ill or that they were correct to obtain medical care. Physicians were significantly more likely to perceive parents as expecting antibiotics if they used certain communication behaviors. This study revealed an incongruity between parents' reported expectations, their communication behaviors, and physicians' perceptions of parents' expectations. PMID- 12585993 TI - Clinical inquiries. Which postmenopausal women should be offered combined HRT? PMID- 12585994 TI - Clinical inquiries. Is pneumococcal vaccine effective in nursing home patients? PMID- 12585995 TI - Clinical inquiries. Does cranberry juice prevent or treat urinary tract infection? PMID- 12585996 TI - Clinical inquiries. Are drug therapies effective in treating Bell's palsy? PMID- 12586006 TI - [Comparison between two 13C-urea breath tests for the diagnosis of Helicobacter pylori infection: isotope ratio mass spectrometer versus infrared spectrometer]. AB - OBJECTIVES: To compare the accuracy of the breath test using the isotope ratio mass spectrometer (IRMS) versus the nondispersive isotope-selective infrared spectrometer (NDIRS) in the diagnosis of Helicobacter pylori infection. METHOD: Multicenter study in 4 Spanish hospitals. One group of dyspeptic patients who had not undergone prior eradication therapy and another group of patients with gastric ulcer or gastrointestinal bleeding due to gastroduodenal ulcer receiving H. pylori eradication therapy were included in the study. A reference standard based on histology and the rapid urease test was used. The breast test (TAU-KIT, Isomed S.L., Madrid, Spain) was performed with citric acid and 100 mg of 13C urea. Samples of expired air were collected in tubes and bags for reading with the IRMS (ABCA, PDZ, Crewe, Manchester, England) and the NDIRS (UBiT-IR200, Otsuka Electronics, Co, Osaka, Japan), respectively. The endoscopist, pathologist and person responsible for reading the urease test and both breath tests were blinded to the results of the other diagnostic methods. RESULTS: Forty-one patients were included. The prevalence of H. pylori was 26%. No differences were found on comparing the mean values obtained with the IRMS and the NDIRS: 13 (standard deviation) (24) and 14 (25) delta units, respectively. The area under the ROC curve for the IRMS and the NDIRS was 0.96. The diagnostic accuracy for the best cut-off point with the IRMS and the NDIRS was, respectively: sensitivity (90 and 100%), specificity (96 and 89%), positive predictive value (90 and 77%), negative predictive value (96 and 100%), + likelihaod ratio (25 and 9.3) and (0.1 and 0). A close correlation was found between the values of the IRMS and those of the NDIRS (lineal regression equation, Y = 1.1 + 1.004. X; r = 0.97). CONCLUSION: Both the spectrometers used to evaluate the breath test, the IRMS and the NDIRS, offer a high degree of accuracy in the diagnosis of H. pylori infection. PMID- 12586008 TI - [Collagenous colitis and small intestinal villous atrophy: manifestations of the same disorder?]. AB - We describe the case of a 64-year-old woman with a previous diagnosis of celiac sprue and no clinical or histological response to gluten withdrawal. The patient presented a history of longstanding recurrent watery diarrhea and was found to have collagenous colitis after further investigation of her diarrhea. Immunological study was incompatible with celiac disease and no other cause of villous atrophy was found. We suggest that this patient may have a separate disease entity unrelated to celiac sprue and consisting of a pan-intestinal inflammatory disorder characterized by the combination of a chronic inflammatory infiltrate in the small and lower bowel together with a subepithelial collagenous band in the colon. PMID- 12586007 TI - [Endoscopic dilatation of caustic esophageal strictures]. AB - OBJECTIVE: To evaluate the results of endoscopic dilatation in caustic esophageal strictures and to analyze the factors associated with a favorable response. PATIENTS AND METHOD: We performed a retrospective study of 33 patients who underwent dilatation with Savary bougie between 1989 and 2001. Response to initial dilatation and outcome during follow-up were analyzed. RESULTS: The mean age was 50 years (8-83) and 58% were women. Intake was accidental in 29 (88%). In all patients, the caustic substance ingested was alkali. Dilatation was started in the acute phase in 12 patients (36%) and 13 presented inflammatory phenomena adjacent to the stenosis. During initial dilatation, 2 1.6 sessions (2-18) were performed and a favorable response was obtained in 18 patients (54%). At the end of follow-up, 68% of the patients presented satisfactory health status. The comparative study revealed that statistically significant variables for a favorable response to treatment were: accidental alkali intake, instauration of treatment in the chronic phase, absence of inflammatory phenomena, and a small number of initial dilatation sessions. CONCLUSIONS: In our series, more than half the patients with esophageal stenosis due to intake of corrosive alkalis showed a favorable initial response. In these patients with acute-phase stenosis who required a greater number of initial dilatation sessions, endoscopic therapy was less effective. PMID- 12586009 TI - [Intrahepatic percutaneous portosystemic shunt in the treatment of refractory ascites]. PMID- 12586010 TI - [Cancer of the rectum: value of mesorectal excision in the prognosis of the disease]. PMID- 12586011 TI - [Biological therapies in inflammatory bowel disease]. PMID- 12586012 TI - [Modulation of immune response in severe acute pancreatitis. Present and future view]. PMID- 12586013 TI - [New therapeutic perspectives in Barretts esophagus]. PMID- 12586014 TI - [Perceived health as a measure of therapeutic effectiveness in functional bowel disorders]. PMID- 12586015 TI - [Value of antibiotic resistance surveillance systems]. PMID- 12586016 TI - [Haemophilus influenzae: phenotype characteristics of strains isolated in 12 Catalan hospitals over one year]. AB - INTRODUCTION: The purpose of this study was to investigate the antigenic and biochemical characteristics, antibiotic susceptibility, and mechanisms for acquiring resistance of Haemophilus influenzae strains isolated in several Catalan hospitals, to determine the current situation regarding this microorganism in our area. METHODS: Serotype, biotype and betalactamase production, as well as susceptibility to eight antimicrobial agents (ampicillin, cefuroxime, cefotaxime, cefixime, clarithromycin, co-trimoxazole, chloramphenicol and ciprofloxacin) were determined in 497 H. influenzae strains isolated from 1 May 1999 to 30 April 2000 in 12 Catalan hospitals. RESULTS: Among the total, 97.5% of strains were nontypable and 50% of the encapsulated strains were serotype b (all isolated from children under 5 years old). There was a predominance of biotype II, though no age or pathologic tropism was found among any of the biotypes. Our series confirms the previously reported trend to decreasing betalactamase mediated ampicillin resistance in our area, mainly in strains from pediatric patients. More betalactamase negative ampicillin-resistant strains (BLNAR) were isolated in children than in adults. One ciprofloxacin resistant strain was detected. CONCLUSIONS: Infections caused by encapsulated H. influenzae strains are infrequent in our area and the relative importance of serotype b is decreasing. Mechanisms for acquiring ampicillin resistance other than betalactamase production are emerging. Surveillance of ciprofloxacin susceptibility is required to predict therapeutic failures with this quinolone. PMID- 12586017 TI - [Sensitivity of Haemophilus influenzae isolates in Spain to 17 oral antibiotics]. AB - INTRODUCTION: The objective of this study was to assess the sensitivity of 400 strains of Haemophilus influenzae isolated in 21 Spanish laboratories in 1999 to 17 oral antibiotics.Methods. An agar dilution method in HT medium was used for sensitivity testing; interpretation of the results followed NCCLS criteria. RESULTS: Percentages of isolates susceptible to the antibiotics tested were as follows: ampicillin 59.5%, amoxicillin/clavulanate 99.2%, loracarbef 66.2%, cefprozil 70.2%, cefaclor 76.2%, cefuroxime 95%, ceftibuten 100%, cefpodoxime 100%, cefixime 100%, rifampin 99.8%, tetracycline 98.2%, chloramphenicol 99.2%, nalidixic acid 97.5%, ciprofloxacin 100%, trovafloxacin 100%, clarithromycin 74%, and azithromycin 100%. CONCLUSIONS: Geographic distribution of sensitivity rates was not homogeneous for some antibiotics. Around 24% of strains were betalactamase producers, requiring higher MICs of antibiotics such as loracarbef, cefprozil and cefaclor than non betalactamase producers. Nevertheless MICs of ceftibuten, cefpodoxime and cefixime were similar in both betalactamase producers and non-producers. Five strains (1.25%) were beta -lactamase (2), but resistant to ampicillin (MIC > or = 8 mg/L) and to amoxicillin/clavulanic acid (MIC > or = 4/2 mg/L). Only three strains showed intermediate sensitivity to chloramphenicol. These strains and four others were inhibited with > or = 4 mg/L of tetracycline. Only one strain was resistant to tetracycline (MIC: 64 mg/L) and to rifampin (MIC: 256 mg/L). All strains were sensitive to azithromycin (MICs < or = 4 mg/L) and all were sensitive to ciprofloxacin and trovafloxacin (MICs < or = 0.5 mg/L). However, ten strains (2.5%) were resistant to nalidixic acid (MIC > or = 4 mg/L). PMID- 12586018 TI - [Association among anti-CagA antibody detection, antibiotic susceptibility, and peptic ulcer in patients with Helicobacter pylori infection]. AB - INTRODUCTION: The aim of this study was to determine the relationship among antibodies against virulence factors (CagA and VacA), clinical status and primary resistance in dyspeptic patients with Helicobacter pylori infection. METHODS: Ninety-eight adult patients with Helicobacter pylori infection who underwent gastric endoscopy for dyspepsia were studied. Specific serum IgG antibodies against CagA and VacA proteins were detected by Western-blot (Helicoblot 2.0). Minimum inhibitory concentrations of metronidazole, amoxicillin, tetracycline and clarithromycin were determined with the E-test. RESULTS: Thirty-nine patients presented peptic ulcer disease and 59 had non-ulcer dyspepsia. CagA protein was detected in 63 patients, and VacA protein in 52 subjects, and both were significantly associated with peptic ulcers (p 5 0.034 and p 5 0.029, respectively). Susceptibility results showed 38.8% of strains resistant to metronidazole and 10.3% resistant to clarithromycin. No resistance to amoxicillin or tetracycline was found. Susceptibility to clarithromycin was more frequent in ulcer patients than in non-ulcer dyspepsia patients (p 5 0.046). CagA protein was more frequent in patients with clarithromycin-sensitive strains (p < 0.001). CONCLUSION: Antibodies against CagA protein were associated with higher antibiotic susceptibility in patients with ulcers or non-ulcer dyspepsia. Thus, anti-CagA antibody detection could be a useful marker of favorable prognosis with antibiotic treatment. PMID- 12586019 TI - [Hepatitis C virus infection in a first level rural hospital: descriptive study in the decade 1991-1999]. AB - INTRODUCTION: To assess epidemiology, clinical manifestations and prognostic factors in subjects diagnosed with hepatitis C virus (HCV) infection in a first level rural hospital. METHODS: This retrospective study includes 142 patients diagnosed with HCV infection at the Hospital de Llerena, from August 1991 to December 1999. Epidemiological and clinical parameters were collected at a mean of 2.7 years after diagnosis and prognostic factors were analyzed. RESULTS: HCV infection predominated in males (69%) and the mean age of patients was 48.3 6 19.3 years. Mechanisms of transmission included unknown (46.5%), intravenous drug use (39.4%), and transfusions (14.1%). Human immunodeficiency virus coinfection was present in 23% of patients and hepatitis B virus (HBV) coinfection in 5.6%. At the time of diagnosis, 111 patients (78.2%) were asymptomatic; 26 (18.3%) presented with complications of portal hypertension and 5 (3.5%) with extrahepatic symptoms. Ultrasonographic signs of portal hypertension were observed in 32.4% of cases. Hepatocarcinoma was detected in 17 patients (12.0%) and extrahepatic neoplasms in 14 (9.9%). Twenty-eight patients died (19.7%). Independent risk factors for mortality included HBV coinfection (OR 26.9; 95% CI 2.19-331.47), ultrasonographic signs of portal hypertension (OR 11.0; 95% CI 3.38 32.61) and diagnosis of hepatocarcinoma (OR 182.7; 95% CI 14.85-2248.21). CONCLUSIONS: Between 1990 and 1999 in our hospital HCV infection was frequently diagnosed in advanced stages and was associated with high mortality, particularly when ultrasonographic signs of portal hypertension or HBV coinfection were present. PMID- 12586021 TI - [Difuse infiltrative lymphocitosis syndrome in a patient with human immunodeficiency virus infection]. PMID- 12586020 TI - [Fever of intermediate duration]. AB - Fever of intermediate duration (FID) is defined as non-localized fever occurring in the community, lasting from one to four weeks, and having no diagnostic orientation after basic clinical, analytical and radiological evaluations are completed. These may include careful recording of clinical history, physical examination, hemogram, plasma creatinine determination, urinalysis, and chest radiography. The etiology of FID is still not completely defined. In our country, 70% of cases are caused by systemic infectious diseases (rickettsiosis, brucellosis, and mononucleosis syndrome) and 7.7% by localized infections; vasculitis and neoplasms account for less than 2%. In most cases FID can be attended on an outpatient basis, with the guarantee of the easy accessibility. In cases without social support or when there is digestive intolerance or criteria of severe disease, hospital care is required. For the diagnostic approach, it is useful to establish basic rules and to organize the study in several stages, taking into account the most frequent etiologies. The first visit would include elaboration of the clinical history, a hemogram, biochemistry analyses, blood cultures, urine culture, serological studies for the most frequent etiologies, a chest radiograph, and other examinations, as indicated by the clinical history data. The care provided and subsequent diagnostic studies performed will depend on the patient's progression and findings from additional studies. Further works conducted in various geographic settings are necessary to define the complete etiological spectrum of FID. PMID- 12586022 TI - [Fever and headache of 5 days' duration, progressing to psychomotor agitation in 48 hours]. PMID- 12586023 TI - [Oxazolidinones and glycopeptides]. AB - We review the chemical structure, pharmacokinetics, antimicrobial spectrum, mechanisms of action and resistance, clinical use and adverse effects of oxazolidinones (linezolid) and glycopeptides. Oxazolidinones inhibit protein synthesis and glycopeptides cell wall synthesis. Antibacterial spectra cover grampositive pathogens, including multiresistant organisms. Linezolid has a 100% absorption and can be administered orally or intravenously; it is cleared by renal and hepatic routes, thus dose adjustments are not needed in moderate renal or hepatic failure. Oral absorption of glycopeptides is minimal. They are excreted by the kidneys (dose adjustments are needed in renal failure). The main clinical indications are the treatment of infections caused by grampositive pathogens; linezolid is also useful for infections caused by glycopeptide resistant enterococcal infections. Linezolid can cause trombocytopenia when treatment lasted longer than two weeks. The main side effect of vancomycin is its nefrotoxicity and teicoplanin can cause fever. PMID- 12586024 TI - [Plasma exchange support therapy in icterohemorrhagic leptospirosis]. PMID- 12586025 TI - [Septic arthritis due to Leclercia adecarboxylata of probable iatrogenic origin]. PMID- 12586026 TI - [Characterization of a new serotype of Salmonella enterica subsp. enterica: serotype Bata]. PMID- 12586027 TI - [Bacteremia due to Capnocytophaga spp.: two cases]. PMID- 12586028 TI - [The clinical significance of broad-spectrum beta-lactamases]. PMID- 12586029 TI - [Risk factors associated with ceftazidime-resistant Klebsiella pneumoniae infection]. AB - INTRODUCTION: Risk factors associated with ceftazidime-resistant Klebsiella pneumoniae (CAZ-R Kp) infection may vary among hospitals and in the same hospital at different time points. Knowledge of these factors is required to establish suitable infection control programs. METHODS: A case-control study was conducted to assess risk factors for CAZ-R Kp infection. Thirty-two cases were compared with 28 controls admitted to a 200-bed general hospital during 1999 and 2000. RESULTS: In the univariate analysis Kp CAZ-R isolates were significantly associated with nosocomial acquisition (OR 5 17.40), prior antibiotic use (OR 5 14.94), particularly ciprofloxacin use (OR 5 5), and hospitalization stay of more than 6 days (OR 5 6.72). Significantly associated variables in the logistic regression analysis included nosocomial acquisition (OR 5 9.29), prior antibiotic use (OR 5 6.21), and particularly, ciprofloxacin use (OR 5 10.84). CONCLUSIONS: Efforts toward more rational overall antibiotic use and particularly ciprofloxacin use, combined with infection control measures are necessary to decrease the prevalence of CAZ-R Kp in our hospital. PMID- 12586030 TI - [Extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in spanish hospitals (GEIH-BLEE Project 2002)]. AB - INTRODUCTION: To describe demographic aspects of a prospective multicentric nationwide study on Escherichia coli and Klebsiella pneumoniae producing extended spectrum b -lactamases (ESBL) in Spain, between March and June 2000. METHODS: 352 E. coli and K. pneumoniae strains showing a phenotype compatible to ESBL production were collected from 40 Spanish Hospitals. A form with demographic and clinical data was filled for every isolate. Identification and susceptibility testing (microdilution, NCCLS guidelines) were performed in a reference centre. RESULTS. 240 strains (170 E. coli and 70 K. pneumoniae) produced ESBL. E. coli ESBL (+) strains were isolated in 33 out of 40 participating hospitals, whereas K. pneumoniae were isolated in 17 hospitals. E. coli ESBL (+) prevalence ranged from 0% to 2.4% and K. pneumoniae ESBL (+) from 0% to 16.7%. Fifty-one percent of E. coli ESBL (+) were from outpatients, whereas 93% of K. pneumoniae ESBL (+) were from inpatients. E. coli ESBL (+) were mainly collected from Medicine and Surgery wards and K. pneumoniae from ICU and Paediatrics. ESBL (+) strains were isolated more frequently from urine samples. Patients' ages ranged from 0 to 93 years for E. coli (median 58 years) and 0 to 83 years for K. pneumoniae (median: 1 year). Paediatric cases excluded, these median values were 64 and 53 years, respectively. CONCLUSIONS: E. coli and K. pneumoniae ESBL (+) were identificated in 90% of the participating hospitals. Although E. coli was more frequently isolated than K. pneumoniae, production of ESBL was more prevalent in K. pneumoniae. Fifty-one percent of ESBL producing E. coli were isolated from outpatients. PMID- 12586032 TI - [Role of bacteria associated with sexually transmitted infections in the etiology of lower urinary tract infection in primary care]. AB - INTRODUCTION: Urinary tract infections (UTI) are the second most frequent type of infectious pathology treated in primary care clinics. The participation of microorganisms associated with sexually transmitted infection has been reported as a cause of UTI; nevertheless this concept is still controversial. To gather data on this subject, we carried out a search for Gardnerella vaginalis, Ureaplasma urealyticum, Mycoplasma hominis and Streptococcus agalactiae besides the common microorganisms involved in UTI. METHODS: A total of 1507 urine cultures from patients with a clinical diagnosis of low UTI were analyzed. Samples were inoculated onto 5% sheep blood agar and McConkey agar, as well as HBT medium for G. vaginalis, and U9B broth and agar E broth for M. hominis and U. urealyticum. The following parameters were analyzed as possible risk factors: age, sex, pregnancy and diabetes status. RESULTS. There were 436 (28.9%) positive urine cultures. Escherichia coli was isolated in 44.34% of cases. Microorganisms associated with sexually transmitted disease were found in 162 (37%): G. vaginalis (25.7%), U. urealyticum (5.9%), S. agalactiae (3.4%) and M. hominis (2%). UTI were more frequent among the 20 to 40 year-old age group, in women and in diabetic patients. CONCLUSIONS: Microorganisms associated with sexually transmitted disease were found in a large percentage of cultures, indicating the need for studies to clarify their role in the etiology of UTI. PMID- 12586031 TI - [Treatment approach for fungal infections in critically ill patients admitted to intensive care units: results of a multicenter survey]. AB - INTRODUCTION: Two consensus conferences taking place in the United States and Spain were organized to optimize diagnosis and treatment of Candida spp. infections. Among other results, clinical scenarios in which early prescription of antifungal agents is indicated were identified. OBJECTIVE: To determine the criteria followed by physicians for prescribing antifungal agents in critically ill patients in our country and to investigate adherence to the guidelines proposed by the consensus conferences. METHODS: A questionnaire was designed and directed to 4th- and 5th-year residents in intensive care medicine and to specialists in intensive care with training in infectious diseases or other medical areas. Four case reports for which expert consensus indicates early antifungal treatment were included in the questionnaire; 1) recurrent peritonitis secondary to perforation of the digestive tract, with mixed flora including fungi; 2) persistent febrile syndrome in a patient with multiple mucosal fungal colonizations treated with broad-spectrum antibiotics; 3) candiduria and pyuria in a febrile patient; and 4) candidemia. RESULTS: A total of 135 questionnaires from 45 different ICUs were returned (60% response rate). In the candidemia and fungal peritonitis examples, early treatment with antifungal agents was indicated in 100% and 85.9% of responses, respectively, whereas for sepsis with multifocal candidiasis and candiduria associated with pyuria and fever, early treatment was prescribed in only 41.5% and 55.6% of responses, respectively. There were no significant differences in response with regard to degree of training of the physicians surveyed. Fluconazole prescription predominated, mainly at doses of 400 mg/day, in mixed peritonitis, disseminated candidiasis and candiduria, whereas amphotericin B lipid formulations were preferentially indicated in cases of candidemia. Antifungal treatment (early or late) was prescribed in all responses for candidemia, in 95.5% for mixed peritonitis (fungi and bacteria), in 79.5% for multifocal candidiasis in patients with persistent sepsis, and in 77.9% for candiduria with fever and pyuria. CONCLUSIONS: Adherence to recommendations from the consensus conferences was high among intensive medicine specialists, with no differences according to level of training in infectious diseases. PMID- 12586033 TI - [Cyclic rotation of antibiotics. Is all that glitters gold?]. AB - Antibiotic cycling refers to the rotation of antimicrobial agents; that is, one specific agent or class of agents is withdrawn from use during a predefined time period, switched to another, and reintroduced at a later time. This strategy of periodic restriction attempts to reduce the selective pressure that antibiotic agents exert on microbial flora, thereby reducing antimicrobial resistance. Such control of antibiotic use has been proposed as an effective measure for preventing the emergence and spread of resistant pathogens, particularly in areas with high levels of antibiotic pressure. Although the first works on antibiotic cycling were published more than two decades ago, the experience with rotational therapy is limited. Most studies on this subject report intriguing and promising results. Nevertheless, a detailed examination of the literature discloses differences in the objectives proposed, the variables analyzed, and the methods for quantification, making generalization of the results difficult. The association between antibiotic use and emergence of multi-drug resistant pathogens has been extensively demonstrated, but the influence of several factors on the mechanisms of emergence and spread of antibiotic resistance in microorganisms makes it difficult to establish a cause-effect relationship. In this article several methodological considerations are suggested for future studies testing this new antibiotic strategy. PMID- 12586035 TI - [Fever and papular eruption with generalized lymph adenopathy and pharyngeal tonsillar inflammation]. PMID- 12586034 TI - [Foliculitis in a patient with Hodkins disease]. PMID- 12586036 TI - [Aminoglycosides]. AB - Aminoglycosides remain as a efficacious class of antimicrobials, commonly used in the clinical practice. In spite of the existence of several mechanisms of resistance, they continue being active against most of the aerobic Gram-negative bacilli. Currently, although aminoglycosides may be used as monotherapy in the urinary tract infections, they are mainly used in combination with b-lactam antibiotics in severe infections caused by Gram-negative bacilli. The knowledge about the pharmacokinetic and pharmacodynamic parameters of aminoglycosides has suggested their use in an once-daily dosing regimen. This dosing has shown as efficacious as multiple-daily dosing regimen in several studies, which also have shown a trend toward a lower toxicity. Among the adverse events, nephrotoxicity and ototoxicity require a careful evaluation during its administration. PMID- 12586037 TI - [Toxic shock due to Staphylococcus lugdunensis]. PMID- 12586038 TI - [Endocarditis due to Micrococcus luteus]. PMID- 12586039 TI - [Endocarditis due to Corynebacterium pseudodiphtheriticum]. PMID- 12586040 TI - [Laboratory diagnosis of parotitis in the post-vaccination era]. PMID- 12586042 TI - [Asthma: from child to adult]. PMID- 12586043 TI - [Experimental models of asthma. Contributions and limitations]. PMID- 12586044 TI - [Primary care physicians' and patients' degree of satisfaction with a new respiratory medicine health care program]. AB - BACKGROUND: No ideal model for coordinating primary care and specialized respiratory medicine has been established. OBJECTIVE: To evaluate primary care physicians' and patients' degree of satisfaction with a new program for coordinating respiratory medicine and primary care. METHOD: A questionnaire survey of the level of satisfaction with a program combining consultation and direct patient care. The study was carried out at a primary care facility whose staff comprised 31 general practitioners. The respiratory medicine staff consisted of one physician and two nurses from the reference hospital. A questionnaire to determine the physicians' and patients' satisfaction with the program asked for a) overall evaluation on a scale of 0 to 10; b) evaluation on a scale naming five levels from "very bad" to "very good", and c) comparison of the new program with previous ones. RESULTS: One thousand one hundred seventy patients were seen in 12 months; 663 (57%) received a first visit, 400 (34%) received a second visit, and 107 (9%) received emergency care. The level of satisfaction was 7.8 (1.2) among physicians and 8.7 (1.5) among patients (p = 0.030); 97% of the physicians and 94% of the patients described the program as "very good" or "good". According to 100% of the physicians and 64% of the patients, the new program was better than the old one (p = 0.110). CONCLUSIONS: The coordinated care plan proposed is accepted by most physicians and patients and uses primary respiratory medicine resources more efficiently. PMID- 12586045 TI - [Assessment of nurses' understanding of tuberculin testing at a general hospital]. AB - BACKGROUND: The last ten years have seen a resurgence of tuberculosis, yet little information is available about the implementation of tuberculin testing and consistency in its use and interpretation. OBJECTIVE: To evaluate knowledge of tuberculin testing among hospital nurses. MATERIAL AND METHOD: A questionnaire on various aspects of the tuberculin test was administered to registered nurses assigned to various services of our university hospital. The questions were grouped by sections (techniques for administering the test, reading the results and interpreting them). RESULTS: One hundred thirty-five nurses were surveyed; 127 (94%) answered all the questions. Overall, only 42% responded to 10 or more of the 14 items on the questionnaire. By sections, questions related to technique of administration, reading the tuberculin test result and interpreting it were answered correctly by 46%, 11% and 7% of the subjects, respectively. Only two variables were independently related to the number of correct responses: 1) working on a medical ward at the time of the survey or 2) administering the test more than once a week. CONCLUSIONS: This study confirms that hospital nurses' understanding of tuberculin testing is unacceptable. A team of professionals skilled in tuberculin testing should be designated at every level of the health care system to assure the reliability of tests performed for clinical or research purposes. PMID- 12586046 TI - [Asthma patients' perception of dyspnea during acute bronchoconstriction]. AB - The aim of this study was to analyze variations in the perception of acute bronchial obstruction among asthmatics in our practice and to try to define the variables that influence inter-individual differences. We studied 153 asthmatics in stable condition, using a Borg scale to measure dyspnea perceived during a histamine bronchial challenge test. To study individual perception we analyzed both absolute magnitude of perception of dyspnea on the Borg scale when forced expiratory volume in 1 second (FEV1) fell 20% (perception score 20-PS20) and the mathematical difference between PS20 and baseline dyspnea (change in Borg, CB). The results were as follows. 1) The factors that affected PS20 according to multiple linear regression were anxiety, baseline dyspnea and the provocative concentration required to produce a 20% fall in FEV1 (PC20). 2) Within each level of asthma severity, there were differences in dyspnea perception when FEV1 fell 20% (analysis of variance of repeated measures), such that patients with mild asthma and no bronchial obstruction perceived more change in dyspnea. 3) CB during the bronchial challenge test distinguished four ways of perceiving dyspnea: 15% were dysperceivers, 13% were hypoperceivers, 48% were normoperceivers and 24% were hyperperceivers. 4) Hypoperceivers and dysperceivers were at greater risk of severe exacerbation whereas hyperperceivers requested unnecessary medical consultations. 5) Subjects with poorer quality of life were usually dysperceivers; hyperperceivers were the second most common type among those with poorer quality of life. In conclusion, a large percentage of asthmatics do not appropriately perceive acute bronchial obstruction, and the simplest way to evaluate their perception is to calculate the change in dyspnea (on a Borg scale) during the bronchial challenge test. The manner of perceiving dyspnea can not be predicted beforehand, yet it has a significant impact on the use of medical resources and patient quality of life, among other aspects. PMID- 12586047 TI - [Sequential use of nocturnal pulse oximetry and respiratory polygraphy (AutoSet) for diagnosing sleep apnea/hypopnea syndrome in high risk patients]. AB - OBJECTIVE: To construct an algorithm for the diagnosis of sleep apnea/hypopnea syndrome (SAHS) based on the optimized sequential use of nocturnal pulse oximetry and respiratory polygraphy (AutoSet, AS) in patients suspected of SAHS. METHODS: Simultaneous performance of pulse oximetry and AS in 145 patients highly suspected of SAHS (at least two of the following signs or symptoms: chronic snoring, observations of repeated apneic events, and abnormal daytime drowsiness). SAHS was diagnosed when the apnea/hypopnea index (AHI) was greater than or equal to 10, based on AS findings. The diagnostic algorithm was obtained by calculating the prevalence of SAHS, calculating the best diagnostic cutoff point for a desaturation index greater than or equal to 4% (ID4%) based on the AHI; analyzing the positive predictive value (PPV) and negative predictive value (NPV) for the cutoff and the correlation and agreement between ID4% and the AHI depending on the presence and severity of SAHS. RESULTS: The overall prevalence of SAHS was 83.4%. The best cutoff point was obtained using ID4% greater than or equal to 10 (sensitivity 82%; specificity 83%). The PPV and NPV for the prevalence calculated were 0.95 and 0.29, respectively. The correlation between ID4% and AHI was 0.84; overall agreement was 0.93. For patients who did not have SAHS, the PPV was 0.49, and the NPV was 0.35; both gradually increased in cases of mild, moderate and severe SAHS (to 0.90 and 0.95, respectively, for severe SAHS). Using these data we created a diagnostic algorithm according to which an ID4% greater than or equal to 30 for nocturnal pulse oximetry (sensitivity 72%, specificity 100%) would indicate that AS testing would be unnecessary. An ID4% less than 30, on the other hand, would lead to further testing (AS, or polysomnography in the event of a negative AS), based on the high prevalence of SAHS in our series. CONCLUSIONS: The sequential use of nocturnal pulse oximetry and AS allows substantial savings of polysomnographic testing in groups at high risk of SAHS. PMID- 12586048 TI - [Non-small cell lung cancer. Survival after radiotherapy and prognostic factors]. AB - OBJECTIVE: To analyze the results of radiotherapy for non-small cell lung cancer and identify the clinical and therapeutic variables that might influence prognosis. PATIENTS AND METHODS: In a population base of 109 patients, 87 met the enrollment criteria. The patients received thoracic radiotherapy over the macroscopic tumor volume and lymph drainage areas, using conventional fractions. Systemic cytostatic therapy was also given to 61% of the patients. RESULTS: Staging was as follows: 33% (28/87) I and II, 20% (18/87) IIIA and 47% (41/87) IIIB. The mean radiotherapy dose was 66.7 Gy (95% CI, 65-67). Full clinical remission was achieved by 22% (18/87), half of whom later suffered recurrences. Partial remission was achieved by 26% (21/87). No response was observed in 52% (42/87). Statistically significant differences in actuarial survival rates were found for staging and tumor response. Mean survival for stages I and II was 29 months (95% CI, 19-39), for IIIA it was 22 (95% CI, 16-28) months, and for IIIB 16 months (95% CI, 12-20). The most common complications were cutaneous, esophageal and pulmonary, with a low incidence of grade 3 toxicity (less than 3%) and absence of grade 4. CONCLUSIONS: No differences in survival were observed for age, tumor location or histological type. Local control of the disease decisively influenced patient survival. PMID- 12586049 TI - [Video-assisted lobectomy through the auscultatory triangle for the surgical treatment of bronchopulmonary carcinoma. Preliminary experience]. AB - OBJECTIVE: To describe a technique for video-assisted lobectomy through the auscultatory triangle (VALAT) and to assess early and medium-term outcome of the procedure in patients with non-small cell bronchopulmonary carcinoma in stage I. PATIENTS AND METHOD: Between April 1999 and May 2002 we performed 25 VALAT procedures (24 lobectomies and 1 bilobectomy) in 25 patients with a mean age of 63.6 years (range 39-80). RESULTS: No deaths occurred during or after surgery. Conversion to conventional thoracotomy was necessary in two cases. One patient was re-operated to resolve hemothorax and one developed a bronchopleural fistula that was treated by pleural drainage. The mean hospital stay was 6.3 days. Twenty three patients have been disease-free throughout a mean follow-up period of 11.5 months (range 1-36). The two-year actuarial survival rate (Kaplan-Meier) was 93% 7%. CONCLUSIONS: VALAT is a safe procedure that nevertheless requires specific training. The rate of complications was low in our patient series and mortality was nil. The outcome for patients with stage I bronchogenic carcinoma over a two year follow-up period has been comparable to results obtained with conventional thoracotomy. PMID- 12586050 TI - [Acute non-cardiogenic pulmonary edema secondary to hydrochlorothiazide therapy]. AB - Several commonly prescribed drugs can cause acute non-cardiogenic pulmonary edema. A cause-effect relationship is usually difficult to establish because symptoms are not specific. We report a case of pulmonary edema induced by a common diuretic, hydrochlorothiazide. This complication can occur after a first dose of the drug or in patients who have been taking it with no side effects. Edema is due to an idiosyncratic reaction rather than an immune response. The clinical course is usually favorable over the first 24 hours with treatment of blood pressure and respiratory support. Given that severity increases with recurrence, we underline the importance of diagnosis in the first episode. PMID- 12586051 TI - [Chronic cough and fever at the onset of giant cell arteritis]. PMID- 12586052 TI - [Pulse alterations related to the performance of flexible bronchoscopy]. PMID- 12586053 TI - [Pulmonary nodular amyloidosis]. PMID- 12586054 TI - Return of the chalones. AB - Members of the TGFbeta superfamily play many roles in embryonic development and adult tissue homeostasis. Now recent work focused on growth and differentiation factors (GDFs) suggest that these TGFbeta-like molecules may also control organ size and may, in fact, be the long sought after chalones, or negative growth regulators. PMID- 12586055 TI - IRE1: a role in UPREgulation of ER degradation. AB - The unfolded protein response entered the mechanistic realm with the discovery of IRE1 as the key signal transducer in yeast. Although also found in mammals, it appeared to function in assisting the work of other players. The featured studies indicate a separate role for IRE1, and highlight the flexibility that bigger eukaryotes possess in this critical pathway. PMID- 12586056 TI - Neutrophils with a license to kill: permeabilized, not stirred. AB - The neutrophil responds to invading microorganisms in part by assembling the NADPH oxidase complex and producing superoxide radicals. Relatively little is known about the intracellular assembly or activation of the oxidase, but Brown et al. in the January issue of Molecular Cell provide a useful strategy involving permeabilized neutrophils to tackle this question. PMID- 12586057 TI - A new trick for Cyclin-Cdk: activation of STAT. AB - New work in Drosophila demonstrates that cdk4 loss causes phenotypes similar to the loss of JAK/STAT pathway components. Cdk4 overexpression can bypass requirements for JAK but not STAT. These results demonstrate a new function for Cdk4 and a new mode of STAT activation. PMID- 12586058 TI - A structural view of integrin activation and signaling. AB - Integrins connect the matrix to the cytoskeleton and propagate structural order between the two systems. A series of elegant structural papers now provides a compelling explanation of how integrins perform this basic function. PMID- 12586059 TI - Where proteins and lipids meet: membrane trafficking on the move. AB - The compartmental organization of eukaryotic cells has fascinated cell biologists for several decades. Detailed morphological, genetic, and biochemical studies have unraveled astonishingly complex molecular machineries involved in establishing and maintaining organelle identity and cell polarization. Many of the transport steps in the secretory and endocytic pathways are subject to manifold regulatory mechanisms, which in turn are interconnected with a plethora of signaling pathways. It therefore does not seem surprising that the cell biology of intracellular protein and lipid transport continues to thrive. The topics covered at the recent meeting on "Protein Transport in the Secretory Pathway" reflect the enormous complexity of how compartmentalization in eukaryotic cells is achieved. PMID- 12586060 TI - A new view of patterning domains in the vertebrate mesoderm. AB - The musculoskeletal system of vertebrates is derived from the embryonic mesoderm. Its structures are categorized as epaxial or hypaxial based on their adult position and innervation. The epaxial/hypaxial terminology is also used to describe regions of the embryonic somites based on fate mapping of somitic derivatives. However, the adult, functional distinctions are not fully consistent with the changing embryonic environments of mesodermal populations during morphogenesis, and the traditional terminology loses accuracy when used to describe certain mutant phenotypes. Here we describe a new terminology naming two mesodermal environments defined by the lineage of the included cells. We discuss how mutant phenotypes may be better explained by consideration of the embryonic context in which genes take their effect and argue that the recognition of these embryonic territories clarifies description and discussion of the morphogenesis and patterning of the musculoskeletal system. PMID- 12586061 TI - A gradient of JAK pathway activity patterns the anterior-posterior axis of the follicular epithelium. AB - The Drosophila egg develops through closely coordinated activities of associated germline and somatic cells. An essential aspect of egg development is the differentiation of the somatic follicle cells into several distinct subpopulations with specific functions. Here we demonstrate that the graded activity of the Janus kinase (JAK) pathway, stimulated by the Unpaired ligand, patterns the anterior-posterior axis of the follicular epithelium. Different levels of JAK activity instruct adoption of distinct anterior cell fates. Further, the coordinated activities of the JAK/STAT and epidermal growth factor receptor (EGFR) pathways are required to specify the posterior terminal cell fate. We propose that Upd secreted from the polar cells may act as a morphogen to stimulate A/P-derived follicular fates through JAK pathway activation. PMID- 12586062 TI - Cyclin D-Cdk4 and cyclin E-Cdk2 regulate the Jak/STAT signal transduction pathway in Drosophila. AB - The JAK/STAT signal transduction pathway regulates many developmental processes in Drosophila. However, the functional mechanism of this pathway is poorly understood. In this report, we identify the Drosophila cyclin-dependent kinase 4 (Cdk4), which exhibits embryonic mutant phenotypes identical to those in the Hopscotch/JAK kinase and stat92E/STAT mutations. Specific genetic interactions between Cdk4 and hop mutations suggest that Cdk4 functions downstream of the HOP tyrosine kinase. We further show that Cyclin D-Cdk4 (as well as Cyclin E-Cdk2) binds and regulates STAT92E protein stability. STAT92E regulates gene expression for various biological processes, including the endocycle S phase. These data suggest that Cyclin D-Cdk4 and Cyclin E-Cdk2 play more versatile roles in Drosophila development. PMID- 12586063 TI - Cholesterol modification of hedgehog is required for trafficking and movement, revealing an asymmetric cellular response to hedgehog. AB - Hedgehog family members are secreted proteins involved in numerous patterning mechanisms. Different posttranslational modifications have been shown to modulate Hedgehog biological activity. We investigated the role of these modifications in regulating subcellular localization of Hedgehog in the Drosophila embryonic epithelium. We demonstrate that cholesterol modification of Hedgehog is responsible for its assembly in large punctate structures and apical sorting through the activity of the sterol-sensing domain-containing Dispatched protein. We further show that movement of these specialized structures through the cellular field is contingent upon the activity of proteoglycans synthesized by the heparan sulfate polymerase Tout-Velu. Finally, we show that the Hedgehog large punctate structures are necessary only for a subset of Hedgehog target genes across the parasegmental boundary, suggesting that presentation of Hedgehog from different membrane compartments is responsible for Hedgehog functional diversity in epithelial cells. PMID- 12586064 TI - P1/HC-Pro, a viral suppressor of RNA silencing, interferes with Arabidopsis development and miRNA unction. AB - The molecular basis for virus-induced disease in plants has been a long-standing mystery. Infection of Arabidopsis by Turnip mosaic virus (TuMV) induces a number of developmental defects in vegetative and reproductive organs. We found that these defects, many of which resemble those in miRNA-deficient dicer-like1 (dcl1) mutants, were due to the TuMV-encoded RNA-silencing suppressor, P1/HC-Pro. Suppression of RNA silencing is a counterdefensive mechanism that enables systemic infection by TuMV. The suppressor interfered with the activity of miR171 (also known as miRNA39), which directs cleavage of several mRNAs coding for Scarecrow-like transcription factors, by inhibiting miR171-guided nucleolytic function. Out of ten other mRNAs that were validated as miRNA-guided cleavage targets, eight accumulated to elevated levels in the presence of P1/HC-Pro. The basis for TuMV- and other virus-induced disease in plants may be explained, at least partly, by interference with miRNA-controlled developmental pathways that share components with the antiviral RNA-silencing pathway. PMID- 12586066 TI - EGF signaling patterns the feather array by promoting the interbud fate. AB - Feather buds form sequentially in a hexagonal array. Bone morphogenetic protein (BMP) signaling from the feather bud inhibits bud formation in the adjacent interbud tissue, but whether interbud fate and patterning is actively promoted by BMP or other factors is unclear. We show that epidermal growth factor (EGF) signaling acts positively to establish interbud identity. EGF and the active EGF receptor (EGFR) are expressed in the interbud regions. Exogenous EGF stimulates epidermal proliferation and expands interbud gene expression, with a concurrent loss of feather bud gene expression and morphology. Conversely, EGFR inhibitors result in the loss of interbud fate and increased acquisition of feather bud fate. EGF signaling acts directly on the epidermis and is independent of BMP signaling. The timing of competence to interpret interbud-promoting signals occurs at an earlier developmental stage than previously anticipated. These data demonstrate that EGFR signaling actively promotes interbud identity. PMID- 12586065 TI - Chordin is required for the Spemann organizer transplantation phenomenon in Xenopus embryos. AB - We analyzed the Chordin requirement in Xenopus development. Targeting of both chordin Xenopus laevis pseudoalleles with morpholino antisense oligomers (Chd-MO) markedly decreased Chordin production. Embryos developed with moderately reduced dorsoanterior structures and expanded ventroposterior tissues, phenocopying the zebrafish chordino mutant. A strong requirement for Chordin in dorsal development was revealed by experimental manipulations. First, dorsalization by lithium chloride treatment was completely blocked by Chd-MO. Second, Chd-MO inhibited elongation and muscle differentiation in Activin-treated animal caps. Third, Chd MO completely blocked the induction of the central nervous system (CNS), somites, and notochord by organizer tissue transplanted to the ventral side of host embryos. Unexpectedly, transplantations into the dorsal side revealed a cell autonomous requirement of Chordin for neural plate differentiation. PMID- 12586067 TI - In vitro generation of long-term repopulating hematopoietic stem cells by fibroblast growth factor-1. AB - The role of fibroblast growth factors and their receptors (FGFRs) in the regulation of normal hematopoietic stem cells is unknown. Here we show that, in mouse bone marrow, long-term repopulating stem cells are found exclusively in the FGFR(+) cell fraction. During differentiation toward committed progenitors, stem cells show loss of FGFR expression. Prolonged culture of bone marrow cells in serum-free medium supplemented with only FGF-1 resulted in robust expansion of multilineage, serially transplantable, long-term repopulating hematopoietic stem cells. Thus, we have identified a simple method of generating large numbers of rapidly engrafting stem cells that have not been genetically manipulated. Our results show that the multipotential properties of stem cells are dependent on signaling through FGF receptors and that FGF-1 plays an important role in hematopoietic stem cell homeostasis. PMID- 12586068 TI - Coordinate regulation of bacterial virulence genes by a novel adenylate cyclase dependent signaling pathway. AB - Type III secretion systems (TTSSs) are utilized by numerous bacterial pathogens to inject effector proteins directly into host cells. Using a whole-genome microarray, we investigated the conditions and regulatory factors that control the expression of the Pseudomonas aeruginosa TTSS. The transcriptional response of known TTSS genes indicates a hierarchical pattern of expression in which a set of secretion apparatus and regulatory genes is constitutively expressed. Further analysis of genes coordinately regulated with those encoding the TTSS led to the identification of a signaling pathway that originates from a membrane-associated adenylate cyclase and controls TTSS gene expression. Transcriptome analysis of mutants lacking the ability to synthesize cAMP or the cAMP binding protein Vfr implicated this pathway in the global regulation of host-directed virulence determinants, including the TTSS. PMID- 12586069 TI - A time-dependent phase shift in the mammalian unfolded protein response. AB - Unfolded or misfolded proteins in the endoplasmic reticulum (ER) must be refolded or degraded to maintain homeostasis of the ER. The ATF6 and IRE1-XBP1 pathways are important for the refolding process in mammalian cells; activation of these transcriptional programs culminates in induction of ER-localized molecular chaperones and folding enzymes. We show here that degradation of misfolded glycoprotein substrates requires transcriptional induction of EDEM (ER degradation-enhancing alpha-mannosidase-like protein), and that this is mediated specifically by IRE1-XBP1 and not by ATF6. As XBP1 is produced after ATF6 activation, our results reveal a time-dependent transition in the mammalian unfolded protein response: an ATF6-mediated unidirectional phase (refolding only) is followed by an XBP1-mediated bidirectional phase (refolding plus degradation) as the response progresses. PMID- 12586073 TI - Proceedings of the Eighth Biennial International Forum on Ovarian Cancer. Houston, Texas, USA. 2001. PMID- 12586070 TI - The Drosophila myosin VI Jaguar is required for basal protein targeting and correct spindle orientation in mitotic neuroblasts. AB - Asymmetric cell divisions generate cellular diversity. In Drosophila, embryonic neuroblasts target cell fate determinants basally, rotate their spindles by 90 degrees to align with the apical-basal axis, and divide asymmetrically in a stem cell-like fashion. In this process, apically localized Bazooka recruits Inscuteable and other proteins to form an apical complex, which then specifies spindle orientation and basal localization of the cell fate determinants and their adapter proteins such as Miranda. Here we report that Miranda localization requires the unconventional myosin VI Jaguar (Jar). In jar null mutant embryos, Miranda is delocalized and the spindle is misoriented, but the Inscuteable crescent remains apical. Miranda directly binds to Jar, raising the possibility that Miranda and its associated proteins are translocated basally by this actin based motor. Our studies demonstrate that a class VI myosin is necessary for basal protein targeting and spindle orientation in neuroblasts. PMID- 12586075 TI - General keynote: hereditary cancer: lessons from Li-Fraumeni syndrome. PMID- 12586076 TI - Specific keynote: hereditary ovarian cancer: what we know. PMID- 12586078 TI - General keynote: expression profiling and the study of ovarian cancer. PMID- 12586081 TI - General keynote: proteomic patterns in sera serve as biomarkers of ovarian cancer. PMID- 12586079 TI - Specific keynote: genome copy number abnormalities in ovarian cancer. PMID- 12586082 TI - Specific keynote: molecular determinants of angiogenesis in ovarian cancer. PMID- 12586084 TI - General keynote: expression of epidermal growth factor receptor family in transgenic mouse models of human breast cancer. PMID- 12586085 TI - Specific keynote: experimental models of epithelial ovarian carcinogenesis. PMID- 12586087 TI - General keynote: the impact of cancer chemoprevention. PMID- 12586088 TI - Specific keynote: chemoprevention of ovarian cancer: the journey begins. PMID- 12586090 TI - General keynote: cancer screening: translation of principles into practice. PMID- 12586091 TI - Specific keynote: ovarian cancer risk assessment and the potential for early detection. PMID- 12586093 TI - General keynote: molecular therapeutics and pharmocogenomics. PMID- 12586094 TI - Specific keynote: molecular therapeutics in ovarian cancer. PMID- 12586096 TI - General keynote: vaccine strategies for the therapy of ovarian cancer. PMID- 12586097 TI - Specific keynote: immunological therapy for ovarian cancer. PMID- 12586099 TI - General keynote: clinical trial design. PMID- 12586100 TI - Specific keynote: clinical trial design problems in ovarian cancer. PMID- 12586103 TI - Specific keynote: in the shadow of illness-supporting women with ovarian cancer. PMID- 12586102 TI - General keynote: patient support. PMID- 12586105 TI - The granulin-epithelin precursor: a putative new growth factor for ovarian cancer. PMID- 12586106 TI - A step further in understanding the biology of the folate receptor in ovarian carcinoma. PMID- 12586107 TI - The different etiologies of mucinous and nonmucinous epithelial ovarian cancers. PMID- 12586108 TI - The anti-proliferative activity of interferon-gamma on ovarian cancer: in vitro and in vivo. PMID- 12586109 TI - Current status of the molecular characterization of the ovarian cancer antigen CA125 and implications for its use in clinical screening. PMID- 12586110 TI - Marine invertebrates and their conservation. PMID- 12586111 TI - Harmonisation of polychlorinated biphenyl (PCB) analyses for ecotoxicological interpretations of southeast Asian environmental media: what's the problem? AB - Within the southeast Asian region, an issue has arisen about how polychlorinated biphenyls (PCBs) should be analysed in environmental samples for risk assessment purposes. Should PCBs be identified as Aroclors (or other commercial mixtures such as Kanechlors), as homologues, as PCB congeners, or as toxic equivalents? The debate appears to centre around separate disciplines that are involved in environmental measurement and analyses, specifically the reporting needs of the environmental chemists and toxicologists. The choice of PCB measurements is generally made on the basis of what question is being asked. This viewpoint presents a general description of the issue, identifies the differences amongst measurement techniques (significance of analytical results, the application and reliability of data, and general costs), characterises the current status of PCB analyses and reporting in the southeast Asian region (specifically, in Cambodia, China, Hong Kong, India, Indonesia, Korea, Japan, the Philippines, Thailand and Vietnam), and proposes a recommended approach for harmonising analytical and toxicological results from laboratories and research groups who are interested in investigating the fate and effects of PCBs on marine/estuarine biota in the region. PMID- 12586112 TI - Review on toxicity testing with marine macroalgae and the need for method standardization--exemplified with copper and phenol. AB - Toxic effects on macroalgae have been compiled. Eighty-two articles have been found in literature during 1959-2000. A total of 120 substances were investigated using 65 different macroalgae species. About one-third of the tested compounds were organic substances (33%), another third metal-organic substances (35%), and the last third were oils (14%), metals (8%), detergents (7.5%) and other inorganic chemicals (2.5%). Half of the substances were only tested once on a single species. Likewise, toxicity data has only been reported for one chemical tested on a single occasion for about half of the 65 species. Thus little is known about the toxic effects on marine macroalgae. The objectives of the previous studies undertaken varied and therefore the toxicity data was presented in numerous ways, e.g. using different exposure times, temperature, light intensity, light regime, salinity, and nutrients, which makes a direct comparison of the data difficult. This review also shows that many stages in the lifecycle of macroalgae are often more sensitive to toxic substances than other aquatic organisms. Consequently, tests using macroalgae may discover toxicity earlier, which would in turn also protect the fauna. If toxic compounds have a negative affect on the distribution and growth of structurally and functionally dominating macroalgae, there may indirectly be a large and harmful influence on the whole marine coastal ecosystem. For this reason tests on macroalgae should be included in control programs along the coasts. PMID- 12586113 TI - The use of biomarkers in environmental monitoring programmes. AB - The monitoring of biological effects has recently become an integral component of environmental monitoring programmes as a supplement to the commonly used contaminant monitoring. Over the years, many biomarkers have been developed that are claimed to be efficient at providing an early warning of deleterious effects on biological systems and for estimating biological effects due to contaminants. Although biomarkers are potentially useful, they have a number of important limitations. In this paper, we examine some of the key assumptions behind the theory and practice of use of biomarkers, and propose a scheme, which may facilitate decisions by environmental managers as to how and when to use biomarkers in their monitoring programmes. PMID- 12586114 TI - Heavy metal concentrations in growth bands of corals: a record of mine tailings input through time (Marinduque Island, Philippines). AB - The impact of copper mining along the western coast of Marinduque Island was investigated. Historic input of mine tailings in the coastal region was traced through variations in heavy metal concentrations in Porites growth bands. Five samples were collected from three reefs showing different modes and extent of exposure to mine tailings. Baseline metal concentrations in Porites were established using a coral from a reef that is least exposed to contamination. The lowest mean values of Cu (0.7 microg/g), Mn (0.8 microg/g), and Zn (1.0 microg/g) were calculated from annual skeletal bands representing five years of growth. Conversely, a sample from a reef adjacent to an old tailings stockpile displayed consistently elevated metal values in its growth bands. Mean Cu, Mn, and Zn values for this coral are 3.1, 1.0 and 1.8 microg/g, respectively. Corals from the Ihatub reef showed a distinct metal concentration peak in their 1996 growth ring. These peaks coincide with a documented release of mine tailings in the Ihatub area during that year. Other metal peaks observed in coral samples correlate with years of high precipitation which may have resulted in increased sediment transport in the region. The metals are presumed to be mostly bound to the aragonite lattice of the coral skeleton, however, contribution from incorporated detrital materials to the observed metal signal (mainly of Fe) could not be easily discounted. PMID- 12586115 TI - The role of the Rio de la Plata bottom salinity front in accumulating debris. AB - The Rio de la Plata, one of the most important South American estuarine environments, is characterized by a bottom salinity front that generates an ecotone between the river and the estuary. Based on bottom trawls and costal sampling we describe the distribution, types, and amount of debris found in the bottom and shoreline across this front. Plastics and plastic bags were the main debris types in both areas. Concentrations of total debris upriver the front were always significantly higher than downriver the front showing that the front acts as a barrier accumulating debris. Moreover, a large part of debris end ups accumulated in the coastal area upriver the frontal position. This area is particularly sensitive because the coastline encompasses an UNESCO Man and the Biosphere Reserve and a Ramsar site, and due to the ecological significance of the front for many valuable species. PMID- 12586116 TI - Cytochrome P450, acetylcholinesterase and gonadal histology for evaluating contaminant exposure levels in fishes from a highly eutrophic brackish ecosystem: the Orbetello Lagoon, Italy. AB - Biochemical markers and ovarian histology were investigated in prespawning females of grass goby (Zosterisessor ophiocephalus) and grey mullet (Mugil cephalus) collected, respectively, in late spring and summer 2000 in four sites of a highly eutrophic brackish ecosystem of central Italy, the Orbetello Lagoon. Exposure to chlorinated and aromatic hydrocarbons was evaluated in fish livers by the somatic liver index (SLI) and by measuring 7-ethoxyresorufin-O-deethylase (EROD) and benzo(a)pyrene monooxygenase (BaPMO) activities. Acetylcholinesterase (AChE) activity was measured in brain and gills to evaluate exposure to organophosphates (OPs) and carbamates (CBs). The gonad somatic index (GSI) was used to confirm ovarian maturation and ovarian histology was investigated as a potential biomarker for environmental effects. Samples from the Western Basin, near a sewage treatment plant (STP) off the town of Orbetello, showed higher SLI values and higher EROD and BaPMO activities than those collected from the Ansedonia Canal (AC) in the Eastern Basin (p<0.05) and respect to those from reference sites: the Albegna River (AR) Delta for grass goby and the Nassa Canal (NC), connected with the sea, for grey mullet both located in the Western Basin as well. Low brain AChE activity was observed in both species from the reference sites (AR and NC) in association with the presence of anomalies in developing oocytes: unexpectedly small in grass goby and irregular disintegrated cytoplasm in grey mullet. The results indicate that the Western Basin is more polluted than the Eastern Basin particularly in the Orbetello where the sewage treatment plant may be a source of aromatic and chlorinated compounds while the Albegna River and the Nassa Canal may be sources of OPs and CBs. PMID- 12586117 TI - Marine introductions in the Southern Ocean: an unrecognised hazard to biodiversity. AB - This study investigated the potential for transport of organisms between Hobart, Macquarie Island and the Antarctic continent by ships used in support of Antarctic science and tourism. Northward transport of plankton in ballast water is more likely than southward transport because ballast is normally loaded in the Antarctic and unloaded at the home port. Culturing of ballast water samples revealed that high-latitude hitchhikers were able to reach greater diversities when cultured at temperate thermal conditions than at typical Southern Ocean temperatures, suggesting the potential for establishment in the Tasmanian coastal environment. Several known invasive species were identified among fouling communities on the hulls of vessels that travel between Hobart and the Southern Ocean. Southward transport of hull fouling species is more likely than northward transport due to the accumulation of assemblages during the winter period spent in the home port of Hobart. This study does not prove that non-indigenous marine species have, or will be, transported and established as a consequence of Antarctic shipping but illustrates that the potential exists. Awareness of the potential risk and simple changes to operating procedures may reduce the chance of introductions in the future. PMID- 12586118 TI - Floating marine debris in coastal waters of the SE-Pacific (Chile). AB - Herein we report on the abundance and composition of floating marine debris (FMD) in coastal waters of the SE-Pacific (off the Chilean coast) during the austral summer 2002. The observed FMD consisted mainly of plastic material (86.9%). Densities of FMD were highest between 20 degrees S and 40 degrees S, corresponding to the main concentrations of human population and activities. Low densities of FMD were found in the south between 40 degrees S and 50 degrees S (<1 item km(-2)). Generally, the highest densities were recorded in nearshore waters of major port cities (>20 items km(-2)), but occasionally high concentrations of debris were also found 50 km offshore. Densities of FMD in coastal waters of the SE-Pacific are of similar magnitudes as those found in coastal waters or inland seas of highly populated regions in the northern hemisphere, indicating the need for improved regulation and legislation in the countries of the SE-Pacific. PMID- 12586119 TI - Cost-efficient methods for marine pollution monitoring at Casey Station, East Antarctica: the choice of sieve mesh-size and taxonomic resolution. AB - Contaminants from sewage discharge and abandoned waste tips enter the marine environment adjacent to Australia's Casey Station, East Antarctica. To establish cost-efficient methods for benthic pollution monitoring the effects of sieve mesh size (0.5 and 1.0 mm) and taxonomic aggregation (family, order and class) on the description of infaunal assemblages were determined. The abundance and taxonomy of fauna retained on a 0.5 mm sieve after passing through a 1.0 mm sieve were examined in this study. The 1.0 mm sieve fraction [Human impacts and assemblages in marine soft-sediments at Casey Station, Antarctica, Ph.D. thesis, University of New England, 2001] contained 70% of individuals and 94% of taxa when compared to combined abundances on the 1.0 and 0.5 mm sieves. Furthermore, the addition of 0.5 mm data did not increase sampling precision or the statistical power to detect differences between locations. Differences between locations were detected when species were aggregated to the family level however, further aggregation to order and class levels altered the perceived pattern of differences. Marine pollution monitoring of the soft-bottom benthos at Casey Station is most cost effective when using a 1.0 mm sieve and identifying fauna to the family level. This is the first reported comparison of sampling techniques using Antarctic benthos. PMID- 12586121 TI - The loss of the chemical tanker Ievoli Sun in the English Channel, October 2000. AB - A chemical product tanker, The Ievoli Sun, was abandoned and later sank approximately 11 miles NW of Alderney in the English Channel in October 2000. The vessel carried a mixed cargo comprising 4000 tonnes of styrene, and 1000 tonnes each of methyl ethyl ketone and isopropyl alcohol. These chemicals are of low to moderate toxicity to aquatic life, are not persistent and have a low potential for bio-accumulation. Both of the solvents are miscible in water, whilst styrene is practically insoluble and, following release, it will rise to the sea surface and rapidly evaporate. More than 1000 tonnes of styrene are believed to have been lost to sea during the incident, resulting in an aerial plume of vapour which crossed Alderney and episodic contamination of air and water in the immediate vicinity of the wreck site. Analysis of edible tissues using coupled gas chromatography/mass spectrometry demonstrated low-level styrene contamination of crabs recovered from pots laid very close to the wreck site prior to the incident. These concentrations posed no risks to human consumers, and no fishery controls were implemented. The remainder of the styrene and the ship's main bunker fuel were recovered from the wreck on the seabed, whilst the two chemical solvents were released to the water column in a slow, controlled manner. PMID- 12586120 TI - Horizontal and vertical distribution of PCBs and chlorinated pesticides in sediments from Masan Bay, Korea. AB - Horizontal and vertical distributions of organochlorine compounds (OCs) were determined in sediments from Masan Bay. The concentrations of polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethanes (DDTs), HCB, hexachlorocyclohexanes (HCHs) and chlordane related compounds (CHLs) in sediments were in the range of 1.24-41.4, 0.28-89.2, 0.02-0.59, nd-1.03, and nd-2.56 ng/g, respectively. The spatial distribution of OCs showed a negative gradient from the inner of the bay to outer part of the bay, indicating that the source of OCs was probably located inside the bay. Compositional pattern of PCB congeners showed a relatively high concentration of high-chlorinated congeners in the inner part of the bay and a relatively low concentration of low-chlorinated congeners in the outer part. In sediment core from Masan Bay maximum concentrations of PCBs and DDTs are observed in the subsurface samples and correspond to an age of early 1980s and late 1960s. The concentration profiles of PCBs and DDTs in sediments of Masan Bay appear to correspond to use of PCBs and DDTs in Korea. PMID- 12586122 TI - Paralytic shellfish toxins in green-lipped mussels, Perna viridis, in Hong Kong. PMID- 12586123 TI - Heavy metals in sediments of the inner shelf off the Gulf of Mannar, South East Coast of India. PMID- 12586124 TI - Perioperative management of oral anticoagulant therapy. PMID- 12586125 TI - Perioperative anticoagulation management in patients who are receiving oral anticoagulant therapy: a practical guide for clinicians. AB - The management of patients who require temporary interruption of oral anticoagulant therapy because of surgery or other invasive procedures is a clinically important topic because of the increasing prevalence of patients who are receiving oral anticoagulants and the availability of low-molecular-weight heparins (LMWHs), which allow out-of-hospital perioperative anticoagulation. The optimal management of such patients has been hampered by the lack of well designed prospective studies investigating the efficacy and safety of different perioperative management strategies. The two main issues that need to be considered in perioperative anticoagulant management is the patient's risk of thromboembolic event when anticoagulant therapy is interrupted and the risk of bleeding that is associated with the surgery or procedure. An assessment of these factors will determine the perioperative management approach. The objectives of this review are to focus on practical issues relating to perioperative anticoagulation and the implementation of a perioperative anticoagulation management approach that can be used in everyday clinical practice. PMID- 12586126 TI - Air travel and risk of venous thromboembolism. PMID- 12586127 TI - Changes of biochemical markers and functional tests for clot formation during long-haul flights. AB - INTRODUCTION: Long-haul flights have been suggested to be associated with an increased risk for thromboembolic events. Until now, changes in the coagulation system during an actual flight have not been investigated. MATERIALS AND METHODS: To explore whether any changes occur in the coagulation system during a real long haul flight molecular markers for coagulation and fibrinolysis were measured in 20 volunteers (10 subjects with a low and 10 with a moderate risk for venous thromboembolism (VTE)) during and after a return flight from Vienna to Washington. In addition, functional measurements of coagulation were performed using activated thrombelastography. RESULTS: Thrombelastographic measurements revealed activation of coagulation in all passengers, who showed an increased activity of FVII and FVIII as well as suppressed fibrinolysis. There was no evidence of a pronounced thrombin and fibrin formation. We did not find any differences between both groups concerning coagulation changes. CONCLUSION: Long haul flights induce a certain activation of the coagulation system. This activated coagulation could be a risk factor for VTE during long-haul flights mainly when other risk factors are present. PMID- 12586128 TI - Correction of INR by prothrombin complex concentrate and vitamin K in patients with warfarin related hemorrhagic complication. AB - We investigated the effect of prothrombin complex concentrate (PCC, median 500 IU) and vitamin K (10-20 mg) or either on blood coagulation and clinical findings in 17 patients with major hemorrhagic complication during warfarin treatment. Their international normalized ratio (INR) at admission was median 2.7 (2.0-above 10.0). In 11 patients treated with PCC and vitamin K, INR decreased to median 1.13 (0.91-1.36) 10 min after the administration with elevation of plasma levels of coagulant factors II, VII, IX, X and protein C.INR decreased abruptly after the administration of PCC without vitamin K in two patients but it increased again 12-24 h after, with decrease of coagulant factors levels. In one of them, a hematoma of the brain enlarged with INR re-increase 12-24 h after the administration. In four patients treated with vitamin K alone, INR decreased slowly from 2.69 (1.03-3.35) to 1.28 (1.25-1.44) 12-24 h after the administration in parallel with gradual increase of the coagulant factors.PCC administration with or without vitamin K seems to be more effective in rapidly correcting increased INR levels than vitamin K treatment without PCC. PCC without vitamin K may result in re-increase of INR and clinical deterioration. PMID- 12586129 TI - Oral-anticoagulant-related intracerebral hemorrhage. AB - BACKGROUND: The characteristics, management and outcomes of patients who suffer intracerebral hemorrhage (ICH) while taking oral anticoagulants (OAC) are relatively unreported. DESIGN: Retrospective cohort study of consecutive cases with ICH associated with OAC. SETTING: A university-affiliated tertiary care hospital in Ontario, Canada. PATIENTS/PARTICIPANTS: 368 charts of individuals with a discharge diagnosis of ICH (ICD-9 code 431) between January 1993 and May 1998 were reviewed. MAIN RESULTS: 20 (5.4%, 95% confidence interval (CI): 3.1 7.7%) of the 368 ICHs occurred in people taking OAC. The median age of patients on OAC was 74 years (S.D.+/-9.8), and 70% (95% CI: 49-91%) were female. The median INR at presentation was 3.4 (intraquartile (IQR) range 2.2-4.4). Nine of 20 (45%) patients had INR values which exceeded the target range. The case fatality rate was 45% (95% CI: 23-67%). Approximately 2.8 years after the initial ICH, 9 of the 11 patients who survived the initial ICH were still alive, and 6 had restarted OAC. CONCLUSIONS: ICH is a serious complication in patients taking OAC, and the case-fatality rate is high. Given the increasing use of OAC in patients with cardiovascular disease, the relative benefits and risks of this therapy must be weighed carefully. PMID- 12586130 TI - Aspirin non-responsiveness as measured by PFA-100 in patients with coronary artery disease. AB - INTRODUCTION: The purpose of the present study was to study the concept of aspirin resistance or non-responsiveness by investigating the response to long term aspirin therapy in patients with a former acute myocardial infarction (AMI). MATERIALS AND METHODS: Patients with an AMI (n=202) randomly assigned to aspirin 160 mg/day (n=71), aspirin 75 mg/day and warfarin (INR 2.0-2.5) (n=58) or warfarin (INR 2.8-4.2) (n=73) were evaluated by the PFA-100(R), biochemical variables and clinical events after a mean treatment period of 4 years. RESULTS: The limit for being an aspirin non-responder was defined as the 95th percentile value in the warfarin alone group (196 s) with the epinephrine cartridge. In patients on aspirin alone 25/71 (35%) were non-responders and on the combination 23/58 (40%). With the adenosine diphosphate (ADP) cartridge only minor differences were found. The levels of thromboxane B(2) in both aspirin groups, in responders as well as in non-responders, were extremely low compared to the warfarin alone group. Evaluating both aspirin groups together (n=129), the levels of soluble P-selectin were significantly higher in non-responders as compared to responders (p=0.012). During the observation period of 4 years with limited number of events, there was a tendency for higher event rates in non-responders as compared to responders (36% vs. 24%, p=0.28). CONCLUSIONS: In our evaluation of the PFA-100(R) a considerable number of post-AMI patients seemed to be non responders to long-term aspirin therapy in doses of 75 and 160 mg/day. Circulating levels of P-selectin were higher in the non-responders. A tendency to higher incidence of clinical events among non-responders was observed. PMID- 12586131 TI - PFA-100 and flow cytometry: can they challenge aggregometry to assess antiplatelet agents, other than GPIIbIIIa blockers, in coronary angioplasty? AB - INTRODUCTION: Platelet response to inhibitors varies widely, leading to a higher risk of abrupt closure events in insufficiently treated-coronary heart disease patients. The aim of this study was to compare, in patients under various antiplatelet regimens, three platelet function assays: aggregometry, PFA-100 and flow cytometry. These assays stand for available tests, as "ready-to-use" device (PFA-100) and sophisticated assay (cytometry). We chose the setting of percutaneous coronary intervention as a standardized procedure to determine which test was appropriate to detect the effect of (1) an aspirin bolus in patients under long-term aspirin treatment, and (2) ticlopidin in case of stent implantation. METHODS: Fifty patients under oral aspirin treatment were randomized to receive a bolus of 500 mg aspirin before angioplasty (n=25). Ticlopidin was given at a 500 mg loading dose in the case of stent implantation (n=38). Platelet function was assessed before, at 2 and 24 h after angioplasty. RESULTS: Considering aspirin antiplatelet effect, the following was observed: (1) a lack of further inhibition after the bolus whatever assay was used and (2) a disagreement between aggregometry and PFA-100 to classify patients as being poor or good aspirin responders (kappa were 0.11 and 0.28 between ADP 4 or 6 microM aggregation, respectively, and PFA-100). Another finding was the good performance of flow cytometry, which evaluated GPIIbIIIa activation, and aggregometry, to detect ticlopidin the day after the loading dose. In contrast, PFA-100 was insensitive to ticlopidin. CONCLUSION: Current assays are not interchangeable to monitor antiplatelet treatment in daily practice. PMID- 12586132 TI - Differential effects of clivarin and heparin in patients undergoing hip and knee surgery for the generation of anti-heparin-platelet factor 4 antibodies. AB - The pathophysiology of heparin-induced thrombocytopenia (HIT) syndrome is mediated via a heterogeneous group of heparin(s)-platelet factor 4 (H-PF4) complexes bound to their antibodies. These anti-H-PF4 (AHPF4) antibodies that are capable of binding to the FcgammaRIIA receptor [cluster of differentiation (CD) 32] on platelets, resulting in platelet activation, widely vary in their specific activities as platelet activation (functionality). Predisposing factors related to specific pathologic conditions may also contribute to the generation of these antibodies and their relative functionality during HIT syndrome. To understand this phenomenon, a sub-study was carried out in patients undergoing elective total hip and knee replacement surgery (ECHOS Study) and who were treated with unfractionated heparin (UFH) and a low-molecular-weight heparin (LMWH; Clivarin). Approximately 600 patients per arm [UFH=7,500 anti-Xa U twice a day (b.i.d.) subcutaneous (s.c.) and clivarin=4200 U once daily (o.d.) s.c.], age >40 years, received prophylactic treatment for a minimum of 11-14 days. Plasma samples were collected at pre-dose, days 2-4, days 11-14 and at follow-up 6-8 weeks after discharge and were analyzed for AHPF4 antibody titers. Functionality of the enzyme-linked immunosorbant assay (ELISA)-positive AHPF4 antibodies to cause platelet activation was tested by 14C-serotonin release assay (SRA). Both UFH and clivarin treatments in orthopedic surgical patients resulted in a progressive generation of AHPF4 antibodies. The relative prevalence/functionality of AHPF4 antibodies in clivarin arm was markedly lower (two- to threefold, p<0.001) as compared to UFH at each time point. Most of the samples in clivarin group were found to be SRA negative, suggesting the presence of AHPF4 antibodies that did not activate platelets (nonfunctional). Within the UFH arm, the relative prevalence/functionality of AHPF4 antibodies was much higher (p<0.002) in knee group compared to the corresponding hip group. This study, for the first time, reports on the elevated levels of AHPF4 antibodies generated by heparin associated with the pathogenesis of knee surgery. Clinical significance of the differential generation of HIT-associated antibodies remains unexplored at this time. PMID- 12586133 TI - Influence of abciximab on the adhesion of platelets on a shielded plasma gradient prepared on polyethylene. AB - INTRODUCTION: Thrombotic effects of biomaterial implants are mediated merely through activation of the platelet glycoprotein IIb-IIIa (GpIIb-IIIa) receptor. Consequently, platelet GpIIb-IIIa receptor inhibitors are successfully used during stent implantation procedures to prevent thrombosis. However, currently a new generation of stents contains surface coating, which changes the surface to more hydrophobic or hydrophilic. This change markedly affects the interaction of platelets and may influence the efficiency of GpIIb-IIIa inhibitors. MATERIALS AND METHODS: To study the influence of the wettability of biomaterials on the effectiveness of abciximab, 5-cm polyethylene gradients with contact angles of 100 degrees to 40 degrees were made by means of glow discharge. Fresh whole blood with or without abciximab was recirculated over this gradient. RESULTS: Inhibition of platelet adhesion by abciximab was maximal, but not complete, on the hydrophobic and moderate hydrophobic part of the gradient, with contact angles of 55 degrees to 90 degrees. Percentage inhibition by abciximab was maximal around 60 degrees. CONCLUSIONS: Intermediate hydrophobicity of currently applied stent materials, such as stainless steel, seems optimal in combination with abciximab. However, on hydrophobic and particularly on hydrophilic materials, abciximab is less effective. PMID- 12586134 TI - PLA1/A2 polymorphism of the platelet glycoprotein receptor IIb/IIIa in young patients with cryptogenic TIA or ischemic stroke. AB - BACKGROUND: The relationship between the PIA2 allele of the Leu(33)Pro polymorphism of glycoprotein IIb/IIIa receptor (GPIIb/IIIa) and ischemic stroke is uncertain. The purpose of this study was to investigate a possible association between the GPIIb/IIIa PIA1/A2 polymorphism and the occurrence of cryptogenic stroke in young patients. METHODS: From a consecutive series of 80 patients aged 45 or less with a recent ischemic stroke or TIA, we selected 45 patients with stroke due to small vessel occlusion or stroke of undetermined etiology (according to the TOAST criteria). Controls were 60 healthy blood donors with a similar age distribution. All patients underwent CT of the brain and were screened for cardiovascular risk factors, cardiac disorders and large vessel disease. The frequency of the PIA2 allele was determined by PCR and Msp1 restriction analysis. RESULTS: Eight patients (16%) and 16 controls (27%) were heterozygous for PIA2 allele. Two patients (4%) were homozygous for PIA2. The relative risk of ischemic stroke associated with PIA2 allele was estimated at 0.8 (95% CI: 0.3-1.9). CONCLUSION: This study does not support the association between the PIA1/A2 polymorphism and cryptogenic stroke or TIA in patients aged 45 or less. PMID- 12586136 TI - Tissue factor regulation and cytokine expression in monocyte-endothelial cell co cultures: effects of a statin, an ACE-inhibitor and a low-molecular-weight heparin. AB - INTRODUCTION: Mounting evidence implies beneficial properties of statins and angiotensin converting enzyme (ACE)-inhibitors beyond those of their original indications in the treatment of coronary artery disease (CAD). Less is known of the mechanisms by which low-molecular-weight (LMW) heparin, also used in unstable CAD, affects the cellular micro-environment. The effects of these drugs in monocyte-endothelial cell co-culture systems have so far been sparsely investigated. MATERIALS AND METHODS: We studied the expression of tissue factor (TF) and the cytokines tumour necrosis factor (TNF)-alpha, interleukin (IL)-6 and IL-10 in a co-culture model with monocytic, vitamin D(3)(vitD(3))-differentiated U-937 cells and human coronary artery endothelial cells (HCAEC), and the effects of the above-mentioned drugs in this system. Cells were co-cultured for 18 h, with or without pre-stimulation of the HCAEC with interferon (IFN)-gamma, and in the presence or absence of simvastatin, enalapril or dalteparin. Analyses of surface tissue factor and intracellular cytokines were done by flow cytometry. RESULTS: Co-culture with activated HCAEC induced tissue factor expression in U 937 cells but not in the endothelial cells. All three drugs significantly reduced tissue factor up-regulation (p<0.001 for each). Co-culture also induced IL-6 expression in U-937 cells and an increase in IL-10 production by HCAEC, none of which was affected by drugs. When cultured separately, both cell types expressed TNF-alpha. This was attenuated in U-937 cells by all three drugs (p<0.001 for each), whereas only enalapril reduced the TNF-alpha content of activated HCAEC (p=0.02). Enalapril also down-regulated the basal expression of IL-6 (p=0.01) and IL-10 (p<0.01) in HCAEC, which simvastatin and dalteparin failed to do. CONCLUSIONS: In this study, we demonstrated for the first time that a statin, an ACE-inhibitor and an LMW-heparin all suppress tissue factor up-regulation in monocyte-endothelial cell co-cultures, thus adding new information regarding the cellular effects of these drugs that may be of importance in the treatment of CAD. PMID- 12586135 TI - The effect of humic acid on the adhesibility of neutrophils. AB - Humic acid (HA), a fluorescent allomelanin, has been implicated as an etiological agent of Blackfoot disease (BFD), a peripheral vascular disease prevailing in the southwest of Taiwan. Clinical and pathological studies reveal that it is similar to atherosclerosis. In this report, the effect of HA on human neutrophils is studied because prolonged and enhanced activation of neutrophils adhered on endothelium may damage the endothelium and initiate the process of thrombosis and vasculitis. METHODS: Neutrophils, treated with various concentrations of HA, were added to culture plates, cultured human umbilical vein endothelial cells (HUVECs), or human umbilical vein endothelium tissue culture for 15 or 30 min. The adhesion of neutrophils was measured qualitatively and quantitatively. The mechanism of neutrophil activation was studied with free radical production and various kinase measurements and their activities' assays. RESULTS: HA was shown to enhance, in a dose-dependent manner, the adhesion of neutrophils on the culture plates, cultured human umbilical vein endothelial cells, and human umbilical vein endothelium tissue culture. The adhesion-enhancing ability of HA is elicited through activation of ERK, P38 mitogen-activated kinase (P38MAPK), and phosphoinositide 3 kinase (PI3K) in neutrophils. HA also induces the NF kappaB activation in neutrophils. CONCLUSION: HA treatment markedly enhanced adhesion and superoxide radical production of neutrophils, the characteristics of activated neutrophils; and all these stimulation effects were blocked by several kinase inhibitors, reflecting the involvement of the ERK, P38MAPK, and PI3K on the activation of neutrophils. The induction of NF-kappaB implied that the consequence of neutrophil activation by HA were similar to other stimulants. The prolonged neutrophil activation will further damage endothelium cell and cause thrombosis, vaculitis, as well as arteriosclerosis. This may partially explain why HA consumption will cause BFD. PMID- 12586137 TI - How to evaluate the influence of blood collection systems on the international sensitivity index. Protocol applied to two new evacuated tubes and eight coagulometer/thromboplastin combinations. AB - Previous reports indicate that the international sensitivity index (ISI) of coagulometer/thromboplastin combinations (measuring systems) may be influenced by blood collection systems. We applied a new protocol to assess the extent with which two new blood collection systems (evaluation tubes) influence the ISI of eight widely used measuring systems. The evaluation tubes were made of plastic and had draw volumes of 1.8 and 2.7 ml; citrate concentration was 0.109 M. Well established collection tubes (from the same manufacturer)-which were made of siliconized glass with draw volume and citrate concentration of 4.5 ml and 0.105 M, respectively-served as control. Plasmas from 20 healthy subjects and 60 patients on oral anticoagulants, collected with the evaluation and control tubes, were tested for prothrombin time (PT) with the eight measuring systems. Plasmas collected with the control tubes were also tested with an international standard for thromboplastin. Results were used to calculate the ISIs of the measuring systems. These were then used to calculate the differences of the crossover international normalized ratio (INR) (i.e., the INR obtained with PT ratios of plasmas collected into the evaluation tubes and the ISI determined with plasmas collected into the control tubes). Overall, the differences were small but measurable, especially for the 2.7-ml draw evaluation tubes and with some of the measuring systems (mean difference, benzo(b)fluoroanthene>pyrene. Special PAH compound ratios, such as phenanthrene/anthracene and fluoroanthene/pyrene, were calculated to evaluate the origin. The collected data suggested that the pyrogenic origins such as motor vehicle exhaust and heavy industry emission were the dominant source of PAH in Korean soils. PMID- 12586161 TI - Chloroform in the environment: occurrence, sources, sinks and effects. AB - The chloroform flux through the environment is apparently constant at some 660+/ 220 Ggyr(-1) (+/-1sigma) and about 90% of the emissions are natural in origin: the largest single source being in offshore sea water (contributing 360+/-90 Ggyr(-1)), with soil processes the next most important (220+/-100 Ggyr(-1)). Other natural sources, mainly volcanic and geological, account for less than 20 Ggyr(-1). The non-natural sources total 66+/-23 Ggyr(-1) and are much better characterised than the natural sources. They are predominantly the result of using strong oxidising agent on organic material in the presence of chloride ion, a direct parallel with the natural processes occurring in soils. Chloroform partitions preferentially into the atmosphere; the equilibrium distribution is greater than 99% and the average global atmospheric concentration has been calculated to be 18.5 pmolmol(-1). Atmospheric oxidation, the principal removal process, is approximately in balance with the identified source fluxes. Chloroform is widely dispersed in the aquatic environment (even naturally present in some mineral waters). Consequently, it is also widely dispersed in the tissue of living creatures and in foodstuffs but there is little evidence of bioaccumulation and the quantities in foodstuffs and drinking water are not problematical for human ingestion at the highest concentrations found. Definitive studies have shown that current environmental concentrations of chloroform do not present an ecotoxicological risk, even to fish at the embryonic and larval stages when they are most susceptible. By virtue of the very small amounts that actually become transported to the stratosphere, chloroform does not deplete ozone materially, nor is it a photochemically active volatile organic compound (VOC). It has a global warming potential that is less than that of the photochemically active VOCs and is not classed as a greenhouse gas. PMID- 12586162 TI - Comparisons of coarse and fine versions of two carbons for reducing the bioavailabilities of sediment-bound hydrophobic organic contaminants. AB - Coarse (whole) and finely ground Ambersorb 1500 and coarse and fine coconut charcoal were compared as to their efficiencies in scavenging organic contaminants desorbed from sediment. Aqueous slurries of a test sediment spiked (1 ppm) with p,p(')-DDE (DDE), 2,2('),5,5(')-tetrachlorobiphenyl (TCB), naphthalene (NAP), or phenanthrene (PHEN), and containing 1% levels of the test carbons were treated by shaking at 35 degrees C while exposed to clusters of low density polyethylene membrane (detox spiders). Controls consisted of spiked sediments and detox spiders but no added carbon of any kind and thus represented unimpeded bioavailabilities (to the spiders). After the treatments--agitation periods from 2.5 to 60 h, depending on contaminant hydrophobicity--the exposed detox spiders were analyzed. The fine carbon of either type was more effective than its coarser variant in obstructing contaminant bioavailabilities. The finer variants of both carbons obstructed the bioavailabilities of NAP and PHEN equally well as did the coarser variants of both. Whole Ambersorb 1500 and coarse coconut charcoal were similarly ineffective in intercepting TCB and DDE. Ground Ambersorb 1500 obstructed virtually all bioavailability of all four contaminants and was far more effective than fine coconut charcoal in intercepting DDE and TCB. An additional experiment compared the effectiveness of ground Ambersorb 1500 and fine coconut charcoal in obstructing the bioavailabilities from sediment of a broad array of spiked organochlorine pesticides. The performance of ground Ambersorb 1500 was again found to be superior; the bioavailable levels of each of the 27 pesticides were markedly lower in the presence of ground Ambersorb 1500 than in the presence of fine coconut charcoal. PMID- 12586163 TI - Pharmaceuticals in STP effluents and their solar photodegradation in aquatic environment. AB - The presence of pharmaceutical compounds in surface waters is an emerging environmental issue. Sewage treatment plants (STPs) are recognized as being the main point discharge sources of these substances to the environment. A monitoring campaign of STP effluents was carried out in four European countries (Italy, France, Greece and Sweden). More than 20 individual pharmaceuticals belonging to different therapeutic classes were found. For six selected pharmaceuticals (carbamazepine, diclofenac, clofibric acid, ofloxacin, sulfamethoxazole and propranolol) present in the STP effluents, the persistence towards abiotic photodegradation was evaluated submitting them to solar experiments at 40 degrees N latitude during spring and summer. Based on experimentally measured quantum yields for the direct photolysis in bi-distilled water, half-life times (t(1/2)) at varying seasons and latitude were predicted for each substance. In salt- and organic-free (bi-distilled) water carbamazepine and clofibric acid are characterized by calculated half-life times of the order of 100 days at the highest latitudes (50 degrees N) in winter, whereas under the same conditions sulphamethoxazole, diclofenac, ofloxacin and propranolol undergo fast degradation with t(1/2) respectively of 2.4, 5.0, 10.6 and 16.8 days. For almost all studied compounds, except propranolol the presence of nitrate ions in aqueous solutions results in a reduction of t(1/2). When present, humic acids act as inner filters towards carbamazepine and diclofenac, and as photosensitizers towards sulphamethoxazole, clofibric acid, oflaxocin and propranolol. PMID- 12586164 TI - Characterisation of the abiotic degradation pathways of oxytetracyclines in soil interstitial water using LC-MS-MS. AB - The fate of oxytetracyclines (OTCs) in soil interstitial water was investigated and the structure of a number of degradation products elucidated in a time related experiment. A previously developed separation method for LC-MS-MS able to base separate and quantify OTC and three of its epimers and degradation products was applied. Compounds detected were 4-epi-oxytetracycline (EOTC) (t(R)=3.0 min), OTC (t(R)=4.4 min), alpha-apo-oxytetracycline (alpha-apo-OTC) (t(R)=11.4 min) and beta-apo-oxytetracycline (beta-apo-OTC) (t(R)=18.4 min). Furthermore, we tentatively identified 4-epi-N-desmethyl-oxytetracycline (E-N-DM-OTC) (t(R)=3.0 min), N-desmethyl-oxytetracycline (N-DM-OTC) (t(R)=3.5), N-didesmethyl oxytetracycline (N-DDM-OTC), 4-epi-N-didesmethyl-oxytetracycline (E-N-DDM-OTC) (t(R)=3.7 and 4.7 min) and 2-acetyl-2-decarboxamido-oxytetracycline (t(R)=8.7) in all samples. Most compounds were only present in trace concentrations (less than 2%) relative to the parent OTC. EOTC was on the other hand formed up to a ratio of 0.6 relative to parent OTC concentration. Only EOTC, E-N-DM-OTC, N-DM-OTC, N DDM-OTC and E-N-DDM-OTC were formed during the time-related experiment. All other compounds were probably only present as impurities in the spiked OTC formulation as they declined in concentration from the start of the experiment. Half-lives (T(1/2), days) of the OTCs in soil interstitial water were in the order of 2 days (EOTC) to 270 days (beta-apo-OTC). PMID- 12586165 TI - Exposure to benzene, toluene, xylenes and total hydrocarbons among snowmobile drivers in Sweden. AB - The exposure to benzene, toluene, xylenes and total hydrocarbons among 25 individuals exposed to exhaust from a snowmobile equipped with a two-stroke engine has been evaluated. Sampling was performed by pumped and diffusive sampling in parallel. There was a relatively bad agreement between the two air sampling methods. The bad agreement can in part be explained by back diffusion of the substances from the samplers, a high face velocity, and deposition of droplets of unburned gasoline onto or in the vicinity of the samplers. The levels of benzene ranged from not detectable (< or =0.01 mgm(-3)) to 2.5 mgm(-3). For toluene, xylenes and total hydrocarbons the exposure was 0.10-12.0, < or =0.05 13.0 and 0.90-273 mgm(-3) respectively. The result from two measurements on individuals travelling on an open sleigh at the rear of the vehicle indicated higher levels of benzene, 0.7-0.8 mgm(-3). Children are often riding as a passenger on a sledge and may thus have a higher exposure than their parents. This study indicates that spare time driving a snowmobile may cause a considerable exposure to benzene. Using a four-stroke engine equipped with a catalyst could reduce the exposure. To reduce the exposure for the passenger on a sleigh an extension of the exhaust pipe may be effective. PMID- 12586166 TI - Kinetics and mechanism of nitrite oxidation by hypochlorous acid in the aqueous phase. AB - The rate coefficient for the reaction of nitrite with hypochlorite and hypochlorous acid has been studied using spectrophotometric measurements. The reaction rate has been determined in a wide range of H(+) concentration (5< or = log[H(+)]< or =11). The kinetics were carried out as a function of NO(2)(-), H(+) and total hypochlorite ([HOCl](total)=[HOCl]+[ClO(-)]+[ClNO(2)]) concentrations. The observed overall rate law is described by: -d[HClO](T)dt=[a[NO(2)( )](2)+b[NO(2)(-)]][H(+)](2)c+d[H(+)]+e[NO(2)(-)][H(+)](2)[HOCl](total)At T=298 K and in Na(2)SO(4) at an ionic strength (I=1.00 M), we obtained using a nonlinear fitting procedure: a=(1.83+/-0.36)x10(7) s(-1), b=(1.14+/-0.23)x10(5) Ms(-1), c=(1.12+/-0.17)x10(-13) M, d=(1.43+/-0.29)x10(-6) M(2) and e=(1.41+/-0.28)x10(3) M where the errors represent 2sigma. According to the overall rate law, a/b=k(1)/k(3), b/e=k(3), c=K(w), d/c=K(a), d=K(a)K(w) and e=K(1)K(a). In Na(2)SO(4) at an ionic strength (I=1.00 M), the values of K(1) and K(a) are (1.1+/-0.1)x10(-4) and 1.28x10(7) M(-1), respectively. A mechanism is proposed for the NO(2)(-) oxidation which involves the reversible initial step: NO(2)( )+HOCl left harpoon over right harpoon ClNO(2)+OH(-) (K(1)), while ClNO(2) undergoes the two parallel reactions: attack by NO(2)(-) (k(1)) and hydrolysis (k(3)). ClNO(2) and N(2)O(4) are proposed as important intermediates as they control the mechanism. The rate coefficients k(1) and k(3) have been determined at different ionic strengths in NaCl and Na(2)SO(4). The influence of the ionic strength and ionic environment has been studied in this work. PMID- 12586167 TI - Characteristics of MnO2 catalytic ozonation of sulfosalicylic acid and propionic acid in water. AB - The characteristics of different types of MnO(2) catalytic ozonation of sulfosalicylic acid (SSal) and propionic acid (PPA) have been investigated in this paper. The experimental results show the dependence of catalytic activity of MnO(2) on organic compounds and the pH of solutions, but it is independent on the type of MnO(2). For example, three types of MnO(2) have not any catalytic activity when ozonation of PPA under the condition of this experiment. All MnO(2) catalytic ozonation of SSal at pH=1.0 have a greater total organic carbon removal than ozonation alone has, however, at pH=6.8 and 8.5, catalytic efficiency is not observed. Furthermore, the batch experimental results indicate that there are no direct relationship between the activity of metal oxide catalytic decomposition of ozone and that of its catalytic degradation of organic compounds. PMID- 12586168 TI - Sorption and desorption hysteresis of organic contaminants by kerogen in a sandy aquifer material. AB - Sorption and desorption hysteresis of 1,2-dichlorobenzene, 1,3,5 trichlorobenzene, naphthalene, and phenanthrene were investigated for the Borden aquifer material with total organic carbon of 0.021% and the isolated natural organic matter (NOM). The isolated NOM is a kerogen type of organic matter with relatively low maturation degree and contained many different types of organic matters including vitrinite particles. The modified Freundlich sorption capacities (logK(')(f) and logK(')(foc)) are very close for the sorption of the four solutes by the isolated NOM and the original sand, respectively. Isotherm non-linearity (n value) and hysteric behaviors are related to solute molecular properties (e.g. K(ow) and molecular size). Kerogen encapsulated by inorganic matrices in the original aquifer may not be accessed fully by solutes. The larger the hydrophobic organic chemical (HOC) (hydrophobic organic contaminant) molecule is, the lower accessibility of the HOC to kerogen. This study disputes widely held hypothesis that sorption to mineral surfaces may play a major role in the overall sorption by low TOC (e.g. 0.1% by mass) geomaterials such as Borden sand. It also demonstrates the importance of the condensed NOM domain, even at very low contents, in the sorption and desorption hysteresis of HOCs in groundwater systems. PMID- 12586169 TI - Development and evaluation of an environmental multimedia fate model CHEMGL for the Great Lakes region. AB - This paper describes the development of a multimedia compartmental model--CHEMGL- which predicts the fate and transport of chemicals in the Great Lakes region and can be used for risk assessment. CHEMGL includes 10 compartments that describe a given region: air boundary layer, free troposphere, lower stratosphere, surface water, sediment, surface soil, vadose soil, groundwater zone, plant foliage and plant root. The model assumes that the compartments are completely mixed and chemical equilibrium between the phases within each compartment is assumed (e.g., suspended solids and biota in water). The attenuation mechanisms include advection, transformation reactions, and diffusive and nondiffusive intermedia transport between compartments. Input parameters include a description of each environmental media, emission rates, and chemical-specific properties and reaction rates. The numerical model results are in good agreement with the analytical solution for an example that examines the fate of benzene. Accordingly, the mathematical and computational components of the model were verified. CHEMGL predicted the concentration of four representative chemicals (atrazine, benzo[a]pyrene, benzene and hexachlorobenzene) in all five basins: Superior, Michigan, Huron, Erie and Ontario. The predicted concentrations fell within one to two orders of magnitude of data reported in the literature. These results suggest that the model is appropriate for estimating the fate and exposure of chemicals for a screening level risk assessment. PMID- 12586170 TI - Comparison of Yatabe-Guilford personality test results in retinitis pigmentosa and glaucoma patients. AB - PURPOSE: To determine the psychological state and personality traits of patients with retinitis pigmentosa (RP) or glaucoma so that a closer and better relationship can be developed with the patients. METHODS: The Yatabe-Guilford personality test was administered to 75 RP patients and 42 glaucoma patients. The latter group included 29 cases of primary open-angle glaucoma, 6 of primary angle closure glaucoma, and 7 of normal-tension glaucoma. The patients were being treated at the Department of Ophthalmology, Chiba University Hospital. As controls, 47 age-matched volunteers were tested. RESULTS: A comparison of these three groups showed that the proportions of patients with cyclic tendency, rhathymia, and lack of cooperativeness traits were significantly higher in the RP group than in the glaucoma group. The lack of cooperativeness value was especially higher in the RP than in the glaucoma group and the control group (Fisher exact test, P <.05). On the other hand, the nervousness value was significantly higher in the glaucoma group than in the RP group and than in controls (Fisher exact test, P <.05). Factors of sex, age, type of disease, corrected visual acuity, and central visual fields did not influence the profiles of the RP and glaucoma groups. CONCLUSION: RP patients had relatively well stabilized personalities and were optimistic, while glaucoma patients tended toward nervousness in comparison with RP patients and controls. PMID- 12586171 TI - Refractive errors and factors associated with myopia in an adult Japanese population. AB - PURPOSE: To investigate the refractive status and factors associated with myopia by a population-based survey of Japanese adults. METHODS: A total of 2168 subjects aged 40 to 79 years, randomly selected from a local community, were assessed in a cross-sectional study. The spherical equivalent of the refractive error was calculated and used in a multiple logistic regression analysis to evaluate the relationships between myopia and possible related factors. RESULTS: The mean (+/- SD) of the spherical equivalent was -0.70 +/- 1.40 diopters (D) in men, and -0.50 +/- 1.44 D in women. Based on +/- 0.5 D cutoff points, the prevalence of myopia, emmetropia, and hypermetropia were 45.7%, 40.8%, and 13.5% in men, and 38.3%, 43.1%, and 18.6% in women, respectively. A 10-year increase in age was associated with reduced risk of myopia [men: odds ratio (OR) = 0.53, 95% confidence interval (CI): 0.44-0.62; women: OR = 0.65, 95% CI: 0.54-0.78]. In men, myopia was significantly associated with higher education (high school: OR = 1.6, 95% CI: 1.1-2.3; college: OR = 2.0, 95% CI: 1.3-3.1) and management occupations (OR = 1.6, 95% CI: 1.0-2.4). For women, high income (OR = 1.5, 95% CI: 1.1-2.2), and clerical (OR = 1.5, 95% CI: 1.0-2.4) and sales/service occupations (OR = 1.7, 95% CI: 1.1-2.6) were also associated with myopia. CONCLUSIONS: The prevalence of myopia in a Japanese population was similar to that in other Asian surveys but higher than in black or white populations. Our study confirmed a higher prevalence of myopia among younger vs. older populations, and a significant association with education levels and socioeconomic factors. PMID- 12586172 TI - A novel nonsense mutation with a compound heterozygous mutation in TGFBI gene in lattice corneal dystrophy type I. AB - PURPOSE: We examined transforming growth factor beta-induced (TGFBI) gene mutations in a family with lattice corneal dystrophy type I. METHODS: The proband was one of the offspring of a consanguineous marriage; 4 affected and 3 unaffected individuals of the family were investigated. Genomic DNA of each case was extracted and used for polymerase chain reaction (PCR). The exon 4, 11, and 12 of the TGFBI gene were directly sequenced. The mutations were confirmed by PCR restriction fragment length polymorphism analysis. RESULTS: There was no significant difference in phenotype between the proband and the other 2 patients, except for progression of the corneal opacity with age. R124C mutation was detected in all affected individuals. In addition, G470X, a novel nonsense mutation, was detected in the proband, resulting in the proband being a compound heterozygote with the TGFBI gene. Her unaffected daughter was found to be heterozygous for G470X. CONCLUSION: It is most likely that the novel nonsense mutation is not pathogenic, and that the mutant keratoepithelin protein with R124C is responsible for the phenotype. PMID- 12586173 TI - Does precipitation reduce tissue staining by indocyanine green dye solutions? AB - PURPOSE: Indocyanine green (ICG) dye precipitates when mixed with certain ophthalmic irrigation solutions. The purpose of this study is to investigate whether this precipitation reduces ICG staining of the anterior lens capsule. METHODS: ICG was diluted with each of the following solutions: BSS Plus, physiological saline, or Opeguard Neo. The products were then examined for green precipitate by light microscopy. The tissue staining capability of each ICG solution was tested at two different concentrations (0.5% and 0.0625%) in porcine lenses, regardless of whether the solution contained precipitate. RESULTS: Green precipitate was observed in both concentrations of ICG solutions diluted with BSS Plus, but not in the solutions diluted with either physiological saline or Opeguard Neo. As assessed with light microscopy, staining of the anterior lens capsule appeared weaker for all 0.0625% ICG solutions compared to the corresponding 0.5% ICG solutions. The precipitate that formed in the 0.5% ICG solution diluted with BSS Plus had little effect on the staining quality of the anterior lens capsule. In contrast, the 0.0625% ICG solution diluted with BSS Plus (w/precipitate) showed weaker staining in the lens capsule compared to the other two 0.0625% ICG solutions (w/o precipitate). CONCLUSION: These results suggest that precipitation of ICG may weaken its capability to stain the anterior lens capsule or other transparent ocular tissues. Therefore, ICG solutions that do not form a precipitate may be more capable of staining tissues at lower concentrations. As for other possibilities to explain the deterioration in staining, the effect of the composition of BSS Plus should also be considered. PMID- 12586174 TI - Study of retinal nerve fiber layer thickness within normal hemivisual field in primary open-angle glaucoma and normal-tension glaucoma. AB - PURPOSE: To determine if nerve fiber layer thickness (NFLT) in glaucoma patients decreases before the development of visual field loss, and if there is a difference in the thinning of NFLT between primary open-angle glaucoma (POAG) and normal-tension glaucoma (NTG) eyes. METHODS: Thirty patients (33 eyes) with POAG and 31 patients (31eyes) with NTG, who had visual field defects localized in either the upper or the lower hemifield verified by Humphrey Field Analyzer (HFA), were measured for NFLT by scanning laser polarimetry (Nerve Fiber Analyzer). Twenty-three normal subjects (23 eyes) matched in refraction and age with the glaucoma patients were recruited as the control group. The total deviation (TD) in each hemifield obtained by HFA and the 180 degrees NFLT of each corresponding hemifield was calculated. Relationships between the TD of the normal or abnormal visual hemifield and the NFLT of the corresponding hemifield were compared among the POAG, NTG, and control groups. RESULTS: The NFLT of the corresponding normal hemifield was decreased both in the POAG group and in the NTG group when compared with the corresponding measurements in control subjects. In POAG eyes, thinning of the NFLT in the corresponding normal hemifield was more remarkable if the TD of the abnormal hemifield was greater, but this tendency was not observed in NTG eyes. CONCLUSION: NFLT is already decreased when the visual field is still normal both in POAG eyes and in NTG eyes. However, the pattern of retinal nerve fiber layer damage in POAG may differ from that in NTG. PMID- 12586175 TI - Retinal changes in Otsuka long-evans Tokushima Fatty rats (spontaneously diabetic rat)--possibility of a new experimental model for diabetic retinopathy. AB - PURPOSE: To study retinal changes in the Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a spontaneously diabetic rat, and evaluate it as a possible new diabetic retinopathy model. METHODS: We examined the retinas of OLETF rats and Long-Evans Tokushima Otsuka (LETO) rats as controls under both a transmission electron microscope (TEM) and a scanning electron microscope (SEM). RESULTS: We detected higher blood sugar level in the OLETF rats than in the LETO rats beginning at 5 months of age. The inner nuclear layers of the retina decreased from 3-4 rows to 2 rows, whereas the photoreceptor cell nuclei decreased from 8 rows to 3-6 rows. Retinal pigment epithelial cells decreased in height, and basal infoldings were poorly developed. Retinal capillary basement membranes were significantly thicker in the OLETF rats than in the LETO rats, and endothelial cell damage was observed. The SEM of vascular corrosion casts of OLETF rats showed tortuosity, microaneurysms, and loop formations. CONCLUSIONS: The development of spontaneous hyperglycemia in OLETF rats was associated with alterations in retina ultrastructure. Changes were similar to those seen in diabetic retinopathy, but did not include either hemorrhages or exudates. The OLETF rat may be a useful animal model for the study of ocular complications in humans. PMID- 12586176 TI - Renin-angiotensin system in proliferative diabetic retinopathy and its gene expression in cultured human muller cells. AB - PURPOSE: To determine if the renin-angiotensin system (RAS) is involved in diabetic retinopathy, we measured the levels of angiotensin-converting enzyme (ACE) and angiotensin II in the vitreous of patients with proliferative diabetic retinopathy (PDR). We also investigated whether the genes of the RAS factors were expressed by cultured human Muller cells. METHODS: Vitreous samples were collected during vitreous surgery from patients with PDR, and from patients with idiopathic macular hole, who served as controls. The concentration of ACE was analyzed, and the level of angiotensin II was quantitatively determined by double antibody radioimmunoassay. In addition, the cDNA, prepared from the mRNA extracted from cultured human Muller cells, was used as a template for reverse transcriptase-polymerase chain reaction with primers selected for renin, angiotensinogen, ACE, and angiotensin receptor type I. RESULTS: The mean concentration of ACE was 0.82 +/- 0.73 IU/L at 37 degrees C in the PDR group, which was significantly higher than the 0.05 +/- 0.07 IU/L at 37 degrees C in the controls. The mean concentration of angiotensin II was 8.77 +/- 4.57 pg/mL in the PDR group, which was significantly higher than the 5.1 +/- 1.7 pg/mL in the controls. mRNA for renin, angiotensinogen, ACE, and angiotensin receptor type I was detected in cultured human Muller cells. CONCLUSIONS: The RAS is locally activated in eyes with PDR, and Muller cells may play a role in this local activation. PMID- 12586177 TI - Immunolocalization of heat shock proteins in the retina of normal monkey eyes and monkey eyes with laser-induced glaucoma. AB - PURPOSE: To examine the expression and localization of heat shock proteins (HSPs) in the retinas of normal and experimentally induced primate glaucoma eyes. These proteins are known to be produced in response to a variety of stresses. METHODS: Experimental glaucoma was induced in the right eyes of three adult monkeys by repeated applications of argon laser to the chamber angle. Immunostaining with a panel of antibodies against HSP 90, 70, 60, 47, and 27 was performed on retinal sections prepared from the normal and glaucomatous monkey eyes. RESULTS: The intensity of immunostaining for HSP 90, 60, and 27 was greatly enhanced in the retinas of glaucomatous eyes. Prominent reactivity was observed in the inner retinal layers, especially in the ganglion cell and nerve fiber layers. The staining intensity for HSP 70 was also moderately increased, while immunoreactivity against HSP 47 remained almost unchanged in glaucomatous retinas. Immunostaining against glial fibrillary acidic protein was increased and the immunolabeling pattern appeared to be identical with that of HSP 90 in glaucoma retinas. CONCLUSIONS: The level of HSP 90, 70, 60, and 27 in primate retinas was increased in experimentally induced ocular hypertension. The differences in expression pattern suggest that each HSP may have its unique role in responding to damage or injury related to intraocular pressure elevation. PMID- 12586178 TI - Measurement of blood flow velocity in feeder vessels of choroidal neovascularization by a scanning laser ophthalmoscope and image analysis system. AB - PURPOSE: To measure the blood flow velocities in feeder vessels of patients with choroidal neovascularization (CNV) secondary to age-related macular degeneration. METHODS: We examined the early images of indocyanine green videoangiography (v ICGA) in 29 patients (29 eyes) with CNV, in whom feeder vessels could be identified by v-ICGA. The v-ICGA images were installed in a personal computer. With original software, optical density measurements were performed for the determination of dye-dilution curves. The flow velocity in a CNV feeder vessel was obtained by analyzing the dye-dilution curves. RESULTS: The mean flow velocity in feeder vessels was 33.8 +/- 32.5 mm/s. The flow velocity in feeder vessels of CNV with a diameter of 1.5 mm or larger was significantly higher than that in feeder vessels of CNV with a diameter smaller than 1.5 mm (P <.05, unpaired t-test). CONCLUSIONS: The flow velocity in CNV feeder vessels can be measured with v-ICGA and a computer-based image analysis system. This system would be useful in the evaluation of choroidal circulation. PMID- 12586179 TI - Neuroimaging analysis of a case with left homonymous hemianopia and left hemispatial neglect. AB - PURPOSE: To correlate the neuro-ophthalmological observations with the magnetic resonance images (MRI) and positron emission tomographic (PET) findings in a case with left homonymous hemianopia and left hemispatial neglect. CASE: A 57-year-old woman underwent surgery for a ruptured anterior communicating artery aneurysm. After she recovered consciousness, it was found that she had left homonymous hemianopia and left hemispatial neglect. Although the hemispatial neglect slowly improved, the homonymous hemianopia persisted. MRI and measurements of cerebral glucose metabolism by 2-fluoro-2-deoxy-D-glucose(FDG)-PET were performed 1 year later. RESULTS: MRI revealed infarctions on the medial surface of the frontal lobe, on the right medial surface of the occipital lobe, and global atrophy of the right cortical hemisphere. FDG-PET disclosed severe glucose hypometabolism in the entire right hemisphere. Glucose metabolism in the right occipital cortex was 61.1% of that in the homologous region on the left side, 62.8% in the right anterior cingulate gyrus, and 93.8% in the temporal-parietal-occipital junction. CONCLUSIONS: The low glucose metabolism in the right visual cortex explains the persistent left hemianopia, and that in the right anterior cingulate gyrus and the right temporal-parietal-occipital junction may be responsible for the left hemispatial neglect. The relatively mild damage in the right temporal-parietal occipital junction explained the recovery of the neglect symptom. Measurements of regional cerebral glucose metabolism by PET are useful for determining the cause of cerebral visual dysfunction and its prognosis after a cerebral lesion. PMID- 12586180 TI - Efficacy of tear eosinophil cationic protein level measurement using filter paper for diagnosing allergic conjunctival disorders. AB - PURPOSE: We investigated the efficacy of the tear sampling method using a filter paper to evaluate eosinophil cationic protein (ECP) levels in patients with allergic disorders. METHODS: Subjects were an allergic group comprising patients with allergic conjunctivitis, vernal keratoconjunctivitis, or atopic keratoconjunctivitis, a Sjogren group comprising patients with secondary Sjogren syndrome, and a control group comprising healthy volunteers. Tears were sampled using the Schirmer Method I and the sample was eluted from the filter paper in 50 microL of elution solution containing phosphate buffer solution with 0.5 M NaCl + 0.1% Tween 20. Then the ECP concentration in the elution sample was determined by the enzyme-linked immunosorbent assay method. RESULTS: Tear ECP level in the allergic group was significantly higher than the levels in the other two groups (P <.01 for the Sjogren group and P <.001 for the control). Furthermore, the tear ECP level of each allergic disease subgroup in the allergic group was significantly higher than that in the control group. CONCLUSIONS: This method of determining tear ECP concentration is useful not only to diagnose allergic conjunctival disorders but also to evaluate their clinical stages. PMID- 12586181 TI - Bilateral iris metastases from prostate cancer. AB - BACKGROUND: Iris metastases of malignant tumors are not common, and most of the cases are unilateral. To our knowledge, there has been no report of bilateral iris tumors metastasized from prostate cancer. CASE: A 71-year-old man with a history of prostate cancer presented with bilateral iris tumors and secondary glaucoma. He had had multiple bone metastases, and had undergone chemotherapy. OBSERVATIONS: Gray-white fleshy tumors spread in the iris of both eyes. Iridocyclitis and secondary glaucoma were present. After external beam radiotherapy, the iris tumors regressed remarkably in volume. CONCLUSIONS: We observed a rare case of bilateral iris tumors, which had probably metastasized from prostate cancer. The tumors were well controlled with conservative therapies including radiation. We believe this article is the first report of bilateral iris tumors metastasized from prostate cancer. PMID- 12586182 TI - Comparison of the intraocular pressure lowering effect of latanoprost and carteolol-pilocarpine combination in newly diagnosed glaucoma. AB - PURPOSE: To evaluate the comparative efficacy of latanoprost monotherapy versus combined carteolol and pilocarpine therapy in patients with newly diagnosed glaucoma. METHODS: Masked randomized prospective trial. This study included 51 patients (64 eyes) with newly diagnosed glaucoma or ocular hypertension. The cases were randomly divided into two treatment groups for administration of latanoprost 0.005% once daily, or of carteolol 2% twice daily and pilocarpine 2% twice daily. Mean diurnal intraocular pressure (IOP) was measured at baseline, week 2, week 4, and month 3 after the beginning of treatment. Changes in mean IOP from baseline to the 3-month visit were determined by an analysis of variance. RESULTS: Mean diurnal IOP values were 25.1 +/- 3.1 mm Hg and 25.5 +/- 2.5 mm Hg at baseline in the latanoprost monotherapy group and in the carteolol-plus pilocarpine group, respectively. Diurnal IOP was significantly decreased from baseline to 3 months in both groups (P <.001). At this time point, latanoprost monotherapy had reduced mean diurnal IOP by 7.2 +/- 2.5 mm Hg (28.7%) and carteolol plus pilocarpine had reduced mean diurnal IOP by 7.4 +/- 2.7 mm Hg (29%). There was no difference between the groups in terms of their IOP reduction effect (P =.51). Decreased visual acuity and twilight vision, blurred vision, and headache were more frequent in the carteolol-plus-pilocarpine group than in the latanoprost group (P <.05). CONCLUSIONS: We concluded that latanoprost monotherapy was at least as effective as the carteolol-pilocarpine combination therapy in reducing mean diurnal IOP in newly diagnosed glaucoma or ocular hypertension. PMID- 12586183 TI - Clinical findings in Japanese patients with Waardenburg syndrome type 2. AB - PURPOSE: To determine the visual characteristics of Japanese subjects with the Waardenburg syndrome type 2. METHODS: The visual functions of 11 albino patients who were identified from the screening of 240 children attending a school for children with a hearing deficit were studied. The ophthalmological examinations included eye position, visual acuity, biomicroscopy, ophthalmoscopy, visual field by confrontation or Goldmann's perimetry, stereoacuity by the Titmus test, and color vision by the Ishihara pseudoisochromatic plates. RESULTS: A combination of congenital sensory deafness and partial ocular albinism without lateral displacement of the lacrimal puncta was observed in 11 (4.6%) of the students with hearing deficit. All these children had sectorial heterochromia irides with local retinal hypopigmentation. Lid deformities were not present. The retinal vasculature was normal, and macular hypoplasty was not found. Other than 1 eye with hyperopic amblyopia, no serious visual disturbance was found in these patients. CONCLUSIONS: The 11 students were classified as having Waardenburg syndrome type 2. None had a critical visual deficit, and all had partial heterochromia irides and retinal hypopigmentation. PMID- 12586184 TI - Health-related quality of life in Behcet patients with ocular involvement. AB - PURPOSE: Health-related quality of life (HRQOL) is an important outcome factor in chronic diseases such as Behcet syndrome. We aimed to investigate the relation of HRQOL to the duration of illness, mental state, and visual acuity of patients with Behcet syndrome. METHODS: We conducted a cross-sectional clinical trial of 45 consecutive Behcet patients with ocular involvement. The control group consisted of an age-, sex-, and education-matched group of 45 healthy individuals. All patients and the controls had been given a complete ophthalmic examination. In addition, they completed a questionnaire comprising the SF-36 Health Survey, Beck Depression Inventory, and Beck Anxiety Inventory. Eight multiple regression analyses were carried out in the patient group to determine whether total anxiety scores, total depression scores, duration of the disease, and visual acuity predicted the dependent variable SF-36 subscales. RESULTS: Using the analysis of variance statistical method, comparisons of the patient and the control groups for depression, anxiety, and the subscales of the SF-36 Health Survey indicated a statistical significance for this battery of tests. CONCLUSIONS: Behcet patients with ocular involvement are susceptible to anxiety and depression when compared to age and sex matched controls. It is important for the ophthalmologist to know that changes in the mental state of his patient may trigger a new ocular attack, and to be aware that these changes may play a critical role in the management and preventive measures for Behcet syndrome. PMID- 12586185 TI - Retinal detachment in the mentally retarded. AB - PURPOSE: To report the clinical features and surgical outcomes of retinal detachment in mentally retarded patients. METHODS: Retrospective review of records of mentally retarded patients who had retinal reattachment surgery at the authors' institutions between February 1994 and February 2000. There were 8 patients with 13 surgically treated eyes. Demographic and clinical data were abstracted from the patients' medical records. RESULTS: The retina of 12/13 (92%) eyes remained reattached after a follow-up ranging from 9 to 78 months. In 6 eyes of the 4 patients whose visual acuity (VA) could be determined, VA improved in 5 eyes of 3 patients. In the remaining 4 patients whose VA could not be determined, improved behavioral patterns suggesting a successful surgical outcome were observed in 3 patients with bilateral retinal detachment, although in one of these patients only unilateral retinal reattachment was achieved. The findings in the eyes in this study agree with the findings in reports on patients with traumatic retinal detachment. CONCLUSIONS: The retinal reattachment rate is fair in the mentally retarded compared with the rate in other segments of the population. Ophthalmological examinations should be provided regularly for mentally retarded persons to keep open the possibility for early sight-improving surgery. PMID- 12586186 TI - Choroidal vascular lesions identified by ICG angiography in a case of familial amyloidotic polyneuropathy. AB - BACKGROUND: To describe a patient with familial amyloidotic polyneuropathy (FAP) whose choroidal vascular lesions were demonstrated dynamically with the use of indocyanine green (ICG) angiography. CASE: A 59-year-old man complained of blurred vision due to vitreal amyloidosis in both eyes. Fundus examination after pars plana vitrectomy showed multiple retinal hemorrhages. OBSERVATIONS: ICG angiography performed after vitrectomy clearly delineated multiple sites of hyperfluorescence indicating tissue staining alongside the major choroidal veins in the lower fundus of his left eye. ICG hyperfluorescence was more evident in the late angiographic phase. Fundus examination and fluorescein angiography revealed no abnormal findings at the corresponding sites of ICG dye leakage. CONCLUSIONS: Choroidal vascular lesions in eyes with FAP were demonstrated in vivo using ICG angiography for the first time. ICG angiography may be very beneficial to evaluate occult choroidal involvement in patients with FAP. PMID- 12586187 TI - Suppression or exacerbation of experimental autoimmune uveoretinitis in Lewis rats by pretreatment with or without an autoantigenic peptide in aluminum hydroxide. AB - PURPOSE: Aluminum hydroxide (Al) is an adjuvant to induce Th2 immune responses. The aim of this study is to investigate the effects of pretreatment with Al on experimental autoimmune uveoretinitis (EAU), a Th1 disease model. METHODS: Lewis rats were pretreated with an uveitogenic peptide (#29)/Al, phosphate-buffered saline (PBS)/Al or PBS. EAU was then induced by immunization of #29 in complete Freund's adjuvant (CFA) with intravenous injection of Bordetella pertussis. Three weeks later, EAU was evaluated histologically and antigen-specific cellular immune responses were assessed. RESULTS: EAU was exacerbated in the PBS/Al group and attenuated in the #29/Al group, compared to the control PBS group. Antigen specific cellular proliferation and interferon-gamma production were augmented in the PBS/Al group and suppressed in the #29/Al group. CONCLUSIONS: Suppression or exacerbation of EAU by the pretreatment in this study is related to inhibition or augmentation of antigen-specific Th1 immunity. PMID- 12586188 TI - A case of acute dacryoadenitis associated with herpes zoster ophthalmicus. AB - BACKGROUND: Acute dacryoadenitis is an uncommon disease. CASE: We present what we believe to be the first reported case of herpes zoster ophthalmicus with the onset of acute dacryoadenitis. OBSERVATIONS: A 30-year-old man complained of severe ocular pain and hyperemia in his right eye. Magnetic resonance imaging (MRI) demonstrated enlargement of the right lacrimal gland and acute dacryoadenitis was diagnosed. Two days after treatment with systemic antibiotics he developed iridocyclitis and skin lesions confined to the first division of the trigeminal nerve; therefore, herpes zoster ophthalmicus was diagnosed. Treatment with acyclovir immediately resolved the ocular pain and swelling of the upper eyelid. MRI conducted in the 4 months after the initial examination showed there was no longer enlargement of the right lacrimal gland. CONCLUSION: Clinicians should be aware that varicella-zoster virus may cause acute dacryoadenitis. PMID- 12586189 TI - Cystoid macular edema associated with latanoprost use in a pseudophakic eye with a history of surgical complications. AB - PURPOSE: To report the development of cystoid macular edema following the use of topical latanoprost in an eye with glaucoma and pseudophakia. CASE: A 73-year-old woman with bilateral pseudophakia and glaucoma had been treated with topical carteolol hydrochloride and isopropyl unoprostone bilaterally. One month later the unoprostone was replaced by latanoprost bilaterally. After 1 month, the patient complained of decreased vision in the right eye, which had a history of surgical complications. RESULTS: Fluorescein angiography showed cystoid macular edema in the right fundus. The edema disappeared 2 weeks after the discontinuation of latanoprost. Cystoid macular edema did not develop in the left eye, which had no history of surgical complications. CONCLUSION: These observations suggest that latanoprost may cause cystoid macular edema in an eye with a history of surgical complications. PMID- 12586201 TI - Flavonoid modulation of ionic currents mediated by GABA(A) and GABA(C) receptors. AB - The modulation of ionotropic gamma-aminobutyric acid (GABA) receptors (GABA-gated Cl(-) channels) by a group of natural and synthetic flavonoids was studied in electrophysiological experiments. Quercetin, apigenin, morine, chrysin and flavone inhibited ionic currents mediated by alpha(1)beta(1)gamma(2s) GABA(A) and rho(1) GABA(C) receptors expressed in Xenopus laevis oocytes in the micromolar range. alpha(1)beta(1)gamma(2s) GABA(A) and rho(1) GABA(C) receptors differ largely in their sensitivity to benzodiazepines, but they were similarly modulated by different flavonoids. Quercetin produced comparable actions on currents mediated by alpha(4)beta(2) neuronal nicotinic acetylcholine, serotonin 5-HT(3A) and glutamate AMPA/kainate receptors. Sedative and anxiolytic flavonoids, like chrysin or apigenin, failed to potentiate but antagonized alpha(1)beta(1)gamma(2s) GABA(A) receptors. Effects of apigenin and quercetin on alpha(1)beta(1)gamma(2s) GABA(A) receptors were insensitive to the benzodiazepine antagonist flumazenil. Results indicate that mechanism/s underlying the modulation of ionotropic GABA receptors by some flavonoids differs from that described for classic benzodiazepine modulation. PMID- 12586202 TI - Identification of cytochrome P450 1A2 as enzyme involved in the microsomal metabolism of Huperzine A. AB - Huperzine A is a reversible and selective cholinesterase inhibitor approved for the treatment of Alzheimer's disease. To identify which cytochrome P450 (CYP) isoenzymes are involved in the metabolism of Huperzine A, an in vitro study was performed with rat liver microsomes and immunoinhibition and chemical inhibition methods. Huperzine A metabolism was analyzed with high-performance liquid chromatography (HPLC) and expressed as Huperzine A disappearance rate. Result showed that 76.2% of Huperzine A metabolism was inhibited by CYP1A2 antibody and 17.8% by CYP3A1/2 antibody. Inhibitory effects produced by CYP2C11 and 2E1 antibodies were minor. The CYP1A2 substrate phenacetin showed an inhibitory effect of 70.3%. In conclusion, Huperzine A metabolism in rat liver microsomes is mediated primarily by CYP1A2, with a probable secondary contribution of CYP3A1/2. CYP2C11 and 2E1 are likely not involved in Huperzine A metabolism. PMID- 12586203 TI - Protective effect of the antipsychotic drug zotepine on dizocilpine-induced neuropathological changes in rat retrosplenial cortex. AB - An atypical antipsychotic drug, zotepine, which is pharmacologically and clinically related to clozapine, has unique therapeutic effects on patients with schizophrenia. It has been demonstrated that clozapine blocks neurotoxicity in the rat retrosplenial cortex induced by administration of the noncompetitive N methyl-D-aspartate (NMDA) receptor antagonist dizocilpine ((+)-MK-801). We examined whether or not zotepine has the ability to block neurotoxicity in the rat retrosplenial cortex induced by administration of dizocilpine. Female Sprague Dawley rats were injected intraperitoneally (i.p.) with vehicle (1 mg/kg), zotepine (5, 10 or 20 mg/kg) or clozapine (20 mg/kg). Fifteen minutes later, animals were injected intraperitoneally (i.p.) with vehicle (1 ml/kg) or dizocilpine (0.5 mg/kg). Neuropathological changes (neuronal vacuolization) were assessed 4 h after administration of dizocilpine. Immunohistochemical analysis of heat shock protein HSP-70, a marker of reversible neuronal injury, was performed 24 h after administration of dizocilpine. The pretreatment with zotepine (5, 10 or 20 mg/kg) significantly decreased the number of vacuolized neurons in the rat retrosplenial cortex 4 h after the administration of dizocilpine (0.5 mg/kg), in a dose-dependent manner. The potency of zotepine (20 mg/kg) for dizocilpine induced neurotoxicity was similar to that of clozapine (20 mg/kg). Furthermore, similar to the case with clozapine (20 mg/kg, i.p.), zotepine (20 mg/kg, i.p.) significantly attenuated the expression of HSP-70 in the rat retrosplenial cortex induced by dizocilpine (0.5 mg/kg, i.p.). The present study suggests that the neuroprotective effects of zotepine- on dizocilpine-induced neurotoxicity are equipotent to those of clozapine. Based on the NMDA receptor hypofunction hypothesis of schizophrenia, the efficacy of zotepine in this study may partly contribute to the unique therapeutic effects of zotepine in patients with schizophrenia. PMID- 12586205 TI - Therapeutically relevant concentrations of neomycin selectively inhibit P-type Ca2+ channels in rat striatum. AB - The effects of neomycin on voltage-activated Ca(2+) channels (VACCs) were studied by Ca(2+)-dependent K(+)- and veratridine-evoked [3H]dopamine release from rat striatal slices. Neomycin (0.01-1 mM) concentration dependently reduced K(+) evoked [3H]dopamine release (IC(50) approximately 25 microM), producing approximately 98% inhibition at 1 mM. Contribution of N-, P- and Q-type Ca(2+) channels to this neomycin-sensitive [3H]dopamine release was tested by the combined application of 100 microM neomycin and selective Ca(2+) channel blockers. The effects of neomycin combined with 1 microM of omega-conotoxin GVIA (N-type Ca(2+) channels) or with 100 nM of omega-conotoxin MVIIC (Q-type Ca(2+) channels) were additive, excluding involvement of N- and Q-type Ca(2+) channels. However, the combined effects of neomycin with 30 nM of omega-agatoxin-IVA (P type Ca(2+) channels) were not additive, suggesting involvement of P-type Ca(2+) channels in neomycin-induced inhibition of [3H]dopamine release. On the other hand, veratridine-evoked [3H]dopamine release was shown to be mediated by Q-type Ca(2+) channels only. In addition, neither the inhibitor of sarcoplasmic reticulum Ca(2+)-ATPase thapsigargin (500 nM) nor the blocker of sarcoplasmic reticulum ryanodine Ca(2+) channels ryanodine (30 microM) modulate veratridine evoked [3H]dopamine release, suggesting no contribution of intracellular Ca(2+) stores. Neomycin (up to 100 microM) did not affect veratridine-evoked [3H]dopamine release, suggesting that intracellular Ca(2+) stores are not a prerequisite for the action of neomycin. Lack of inhibitory effect of neomycin is taken as additional indirect evidence for the involvement of P-type Ca(2+) channels. In conclusion, therapeutically relevant concentrations of neomycin preferentially block P-type Ca(2+) channels which regulate dopamine release in rat striatum. This block could be responsible for aminoglycoside-induced toxicity. PMID- 12586204 TI - Antidepressant-like actions of DOV 21,947: a "triple" reuptake inhibitor. AB - DOV 21,947 [(+)-1-(3,4-dichlorophenyl)-3-azabicyclo-[3.1.0]hexane hydrochloride] inhibits the reuptake of [3H]serotonin, [3H]norepinephrine, and [3H]dopamine in human embryonic kidney (HEK) 293 cells expressing the corresponding human recombinant transporters (IC(50) values of 12, 23, and 96 nM, respectively). This compound also inhibits [125I]RTI 55 (3beta-(4-iodophenyl)tropane-2beta-carboxylic acid methyl ester) binding to the corresponding transporter proteins in membranes prepared from these cells (K(i) values of 99, 262, and 213 nM, respectively). DOV 21,947 reduces the duration of immobility in the forced swim test (using rats) with an oral minimum effective dose of 5 mg/kg. This antidepressant-like effect manifests in the absence of significant increases in motor activity at doses of up to 20 mg/kg. DOV 21,947 also produces a dose-dependent reduction in immobility in the tail suspension test, with a minimum effective oral dose of 5 mg/kg. The ability of DOV 21,947 to inhibit the reuptake of three biogenic amines closely linked to the etiology of depression may result in a therapeutic profile different from antidepressants that inhibit the reuptake of serotonin and/or norepinephrine. PMID- 12586206 TI - The effects of a peripherally acting cholecystokinin1 receptor antagonist on food intake in rats: implications for the cholecystokinin-satiety hypothesis. AB - The observation that systemic administration of the peptide cholecystokinin (CCK) inhibits food intake in mammalian species has led to the hypothesis that endogenous peripheral CCK released from the small intestine during a meal acts as a satiety factor. It was predicted that if CCK does play an important role in satiety, then systemic administration of a specific CCK receptor antagonist should block the effects of the endogenous peptide released during a meal and increase food intake. The present study was undertaken to test the hypothesis by investigating the effects of the cholecystokinin(1) (CCK(1)) receptor antagonist N-alpha-3'-quinolinoyl-D-Glu-N,N-dipentylamide dicyclohexylammonium (A70104), which is unlikely to cross the blood-brain barrier, on food intake in rats. A70104 (20-200 microg/kg, i.p.) had no any significant effect on the intake of a test meal in rats under different experimental conditions. However, pretreatment of rats with A70104 (50 microg/kg, i.p.) abolished the inhibitory effects of exogenous peripheral CCK (5 microg/kg, i.p.) on food intake. The findings that A70104 had no effect on food intake when administered on its own, but abolishes the suppressant effect of exogenous peripheral CCK, suggest that endogenously released peripheral CCK does not play an important role as a satiety factor in rats. PMID- 12586207 TI - Pharmacodynamic mechanisms of monoclonal antibody-based antagonism of (+) methamphetamine in rats. AB - Our studies examined pharmacokinetic mechanisms involved in high-affinity (K(d) approximately 11 nM) monoclonal antibody-based antagonism of (+)-methamphetamine induced locomotor effects. Male rats received (+)-methamphetamine (0.3, 1, or 3 mg/kg i.v.) followed 30 min later by saline or anti-(+)-methamphetamine monoclonal antibody. All groups received a constant dose of monoclonal antibody that was equimolar in binding sites to the body burden of a 1 mg/kg i.v. (+) methamphetamine dose 30 min after administration. The monoclonal antibody antagonized locomotor effects due to 0.3 and 1 mg/kg (+)-methamphetamine. In contrast, monoclonal antibody treatment increased locomotor activity due to 3 mg/kg (+)-methamphetamine. We also investigated the serum and brain pharmacokinetics of (+)-methamphetamine without and with the monoclonal antibody. Rats received (+)-methamphetamine (1 mg/kg i.v.) followed by saline or monoclonal antibody treatment at 30 min. The monoclonal antibody significantly increased serum methamphetamine concentrations and significantly decreased brain methamphetamine concentrations. These data indicate that anti-(+)-methamphetamine monoclonal antibody-induced pharmacodynamics are complex, but are related to time dependent changes in (+)-methamphetamine brain distribution. PMID- 12586208 TI - An endothelium-derived factor modulates purinergic neurotransmission to mesenteric arterial smooth muscle of hamster. AB - The interaction between the endothelium and purinergic perivascular nerves was investigated by measuring the changes in amplitude of excitatory junction potential (EJP) of smooth muscle cells in hamster mesenteric arteries (100-350 microm). Uridin-5'-triphosphate (UTP) (100 microM) applied to endothelium-intact preparations evoked a hyperpolarization of 17.0 +/- 0.7 mV (n=46). During this hyperpolarization, the amplitude of electrically evoked EJPs was inhibited to about 50% of that of the control. In endothelium-denuded preparations, UTP (100 microM) neither hyperpolarized the smooth muscle nor inhibited the amplitude of the EJP. Neither a nitric oxide (NO) synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME) (100 microM), nor a cyclooxygenase inhibitor, indomethacin (1 microM), had an effect on the UTP-evoked hyperpolarization and inhibition of the electrically evoked EJP. The UTP-evoked membrane hyperpolarization and inhibition of the EJP amplitude was antagonized by the P2Y receptor antagonist, cibacron blue (100 microM). Endothelium-derived hyperpolarizing factor (EDHF) mediated hyperpolarization was inhibited by either adventitial or intimal application of apamin (0.1 micro and charybdotoxin (0.1 microM). However, the EJP inhibition was still present. In apamin- and charybdotoxin-treated preparations, focal application of adenosine 5'-triphosphate (ATP) (10 mM) evoked a depolarization of 15.5 +/- 1.3 mV (n=15). This postjunctional response was not modified by UTP (15.3 +/- 1.7 mV, n=4, P>0.05). These results suggest that exogenously applied UTP activates P2Y receptors of endothelium to release endothelium-derived factors, which in turn inhibit ATP release from purinergic nerves. PMID- 12586209 TI - Role of nitric oxide and free radicals in cardioprotection by blocking Na+/H+ and Na+/Ca2+ exchange in rat heart. AB - Inhibition of Na(+)/H(+) (NHE) and Na(+)/Ca(2+) (NCE) exchangers prevents myocardial ischemia/reperfusion injury by preventing cardiomyocyte Ca(2+) overload. We hypothesized that it may influence ischemic/reperfused myocardium also indirectly by preventing endothelial Ca(2+) accumulation, and thereby by attenuating reperfusion-induced formation of nitric oxide (NO) and/or oxygen free radicals. Langendorff-perfused rat hearts were subjected to 30-min ischemia and 30-min reperfusion. Myocardial outflow of NO (nitrite+nitrate) and hydroxyl radical (*OH, salicylate method), and functional recoveries were followed during reperfusion. In all groups, there was a transient rise in NO and *OH outflow upon reperfusion. An inhibitor of NHE, cariporide (10 microM) [(4-Isopropyl-3 methylsulfonyl-benzoyl)-quanidine methanesulfonate], and an inhibitor of the reverse mode of NCE, KB-R7943 (5 microM) (2-[4-(4 Nitrobenzyloxy)phenyl]ethyl]isothiourea mesylate), decreased NO and *OH formation, reduced contracture, and improved the recovery of mechanical function during reperfusion, compared to the untreated hearts. The formation of NO was reduced by 40% by 100 microM N(G)-methyl-L-arginine acetate salt (L-NMMA, NO synthase inhibitor), and not affected by 50 microM L-NMMA. *OH formation, contracture, and the functional recoveries were affected neither by 50 nor by 100 microM L-NMMA. Also, the effects of cariporide and KB-R7943 were unaffected by 100 microM L-NMMA. This study shows for the first time that the inhibition of NHE and NCE attenuates post-ischemic myocardial formation of NO and *OH, suggesting that prevention of Ca(2+) overload is cardioprotective via these mechanisms. The results indicate, however, that NO synthase pathway did not interfere with the protection afforded by NHE or NCE in our model. PMID- 12586210 TI - Selegiline attenuates cardiac oxidative stress and apoptosis in heart failure: association with improvement of cardiac function. AB - We have shown recently that selegiline exerts a cardiac neuroprotective effect in chronic heart failure. Since selegiline has an antioxidant antiapoptotic effect, we proposed to determine whether selegiline attenuates cardiac oxidative stress and myocyte apoptosis in chronic heart failure by modulating Bcl-2 and Bax protein expression, and whether the effects are associated with the improvement of cardiac function. Rabbits with rapid cardiac pacing (360 beats/min) and sham operation without pacing were randomized to receive oral selegiline (1 mg/day) or placebo for 8 weeks. Echocardiography was used to measure left ventricular fractional shortening. After 8 weeks of treatment, animals were studied for arterial norepinephrine and left ventricular systolic function (fractional shortening and dP/dt), and were then sacrificed for measuring the stable oxidative product of myocardial mitochondrial DNA (mtDNA) 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxo-dG), myocyte apoptosis by monoclonal antibody to single stranded DNA, and Bcl-2 and Bax protein expression by Western blot and immunohistochemistry. Rapid cardiac pacing increased plasma norepinephrine, cardiac oxidative stress and myocyte apoptosis, reduced Bcl-2 and the Bcl-2 to Bax ratio. These changes were associated with decreased left ventricular fractional shortening and dP/dt. Selegiline treatment in chronic heart failure animals reduced plasma norepinephrine, cardiac oxidative stress and myocyte apoptosis, prevented the changes of Bcl-2 and Bcl-2 to Bax ratio, and improved left ventricular fractional shortening and dP/dt. The findings suggest that the reduction by selegiline of myocyte apoptosis is related to the decrease of cardiac oxidative stress and the modulation of apoptotic and antiapoptotic proteins. The antioxidant antiapoptotic effects of selegiline are potentially beneficial in the improvement of cardiac function in chronic heart failure. PMID- 12586211 TI - Cardiovascular and renal effects of cyclooxygenase inhibition in transgenic rats harboring mouse renin-2 gene (TGR[mREN2]27). AB - The present study examined the role of cyclooxygenase-synthetized prostanoids in the pathogenesis of angiotensin-II-induced inflammatory response and vascular injury in transgenic rats harboring mouse renin-2 gene (mREN2 rats). Five- to six week-old, heterozygous mREN2 rats received the following drug regimens for 8 weeks: (1) controls; (2) cyclooxygenase-2 inhibitor (MF-tricyclic [3-(3,4 difluorophenyl)-4-(4-(methylsulfonyl) phenyl)-2(5H)-furanone], 14 mg kg(-1) p.o.); (3) cyclooxygenase-1/cyclooxygenase-2 inhibitor (sulindac, 14 mg kg(-1) p.o.); (4) angiotensin II receptor antagonist (losartan 40 mg kg(-1) p.o.); (5) MF-tricyclic + losartan; (6) sulindac + losartan. Normotensive Sprague-Dawley rats served as controls. mREN2 rats developed pronounced hypertension, cardiac hypertrophy, and albuminuria as compared to normotensive Sprague-Dawley controls. mREN2 rats showed pronounced perivascular inflammation and morphological damage in the kidneys and the heart. Both MF-tricyclic and sulindac further increased blood pressure and albuminuria in mREN2 rats. Neither MF-tricyclic nor sulindac were able to prevent angiotensin-II-induced perivascular inflammation and morphological changes in the heart or in the kidneys. Myocardial and renal cyclooxygenase-2 mRNA expressions were decreased in mREN2 rats, whereas no difference was found in cyclooxygenase-1 mRNA expressions. Sulindac increased both cyclooxygenase-1 and cyclooxygenase-2 gene expressions, whereas MF-tricyclic increased only cyclooxygenase-2 gene expressions. Losartan normalized blood pressure, cardiac hypertrophy, albuminuria, inflammatory response and morphological changes in mREN2 rats, both in the presence and absence of cyclooxygenase inhibitors. Our findings indicate that cyclooxygenase does not play a central role in the pathogenesis of angiotensin-II-induced inflammatory response and vascular injury in mREN2 rats. PMID- 12586212 TI - Cardioprotective actions of an N-terminal fragment of annexin-1 in rat myocardium in vitro. AB - We have previously shown that the glucocorticoid dexamethasone prevents the cardiodepressant actions of interferon-gamma plus lipopolysaccharide in cardiac tissue in vitro. We now demonstrate that an N-terminal fragment of annexin-1 (Ac2 26, 1 microM), a putative mediator of glucocorticoid actions, completely protects against interferon-gamma+lipopolysaccharide-induced depression of the inotropic response to isoprenaline in rat isolated papillary muscles. However, Ac2-26 does not preserve resting contractile function. Fifteen hours incubation with interferon-gamma+lipopolysaccharide also markedly induced mRNA expression (by real time polymerase chain reaction, PCR) of both the nitric oxide synthase 2 (NOS2) isoform of nitric oxide synthase (by 6.7 +/- 1.7-fold, P < 0.01) and cyclo oxygenase-2 (by 3.4 +/- 0.6-fold, P < 0.05) in cardiomyocytes. Pretreatment with Ac2-26 (1 microM) prevented the induction of cyclo-oxygenase-2 mRNA, but not NOS2 mRNA, whereas dexamethasone (1 microM) suppressed the expression of both NOS2 mRNA and cyclo-oxygenase-2 mRNA. Co-incubation of dexamethasone with an anti annexin-1 antibody did not attenuate the suppression of NOS2 mRNA. Thus, Ac2-26 reproduces some, but not all, of the cardioprotective effects of glucocorticoids in vitro in the absence of neutrophils. These protective actions are independent of changes in NOS2 expression. PMID- 12586213 TI - Carboxypeptidase B inhibitors reduce tissue factor-induced renal microthrombi in rats. AB - Procarboxypeptidase B (also known as thrombin-activatable fibrinolysis inhibitor) is a recently described plasma zymogen known to be activated by thrombin in plasma. Carboxy-terminal lysine residues from partially degraded fibrin are important for the binding and activation of plasminogen, and carboxypeptidase B, an active form of procarboxypeptidase B, has been shown to inhibit fibrinolysis by eliminating these residues. The present paper investigates the effects of carboxypeptidase B inhibitors, DL-mercaptomethyl-3-guanidinoethylthiopropanoic acid (MGPA) and potato-derived carboxypeptidase inhibitor (CPI), on tissue factor (TF)-induced microthrombosis in rats. Intravenous injection of MGPA (3 mg/kg and higher) or CPI (0.3 mg/kg and higher) after microthrombi formation dramatically attenuated TF-induced glomerular fibrin deposition with an increase in plasma levels of D-dimer. These results indicate that carboxypeptidase B inhibitors can enhance endogenous fibrinolysis and reduce thrombi in the TF-induced microthrombosis model after systemic administration even after thrombi formation. PMID- 12586214 TI - Preventive effect of zelandopam, a dopamine D1 receptor agonist, on cisplatin induced acute renal failure in rats. AB - To elucidate the role of peripheral dopamine D1 receptors in cisplatin-induced acute renal injury, effect of zelandopam (YM435, (-)-(S)-4-(3,4-dihydroxyphenyl) 7,8-dihydroxy-1,2,3,4-tetrahydroisoquinoline hydrochloride hydrate), a selective renal dopamine D1 receptor agonist, on cisplatin-induced acute renal failure in rats was studied. Rats were divided into six groups: control, cisplatin and cisplatin plus zelandopam (30, 100, 300 mg/kg p.o. twice, 75 and 15 min before cisplatin injection) or the free radical scavenger CV-3611 (2-O-octadecylascorbic acid, 10 mg/kg p.o., 75 min before cisplatin injection) treated groups. Rats received intraperitoneal injection of cisplatin at a dose of 5 mg/kg. Four days after cisplatin injection, plasma creatinine, blood urea nitrogen and body weight were measured and the kidneys were removed for histological examination. Cisplatin induced acute renal failure characterized by the increases in plasma creatinine and blood urea nitrogen with tubular damage, and decreased body weight. Zelandopam dose-dependently prevented all these changes. The free radical scavenger CV-3611 significantly attenuated a decrease in body weight and renal dysfunction without reducing tubular damage. The present study is the first demonstration for that a selective dopamine D1 receptor agonist is effective in preventing acute renal failure induced by cisplatin. PMID- 12586215 TI - Antiemetic effect of a tachykinin NK1 receptor antagonist GR205171 on cisplatin induced early and delayed emesis in the pigeon. AB - Cisplatin (4 mg/kg, i.v.) induced both early emesis, which appears within the first 8-h period, and delayed emesis, which appears between 8 and 48 h after its administration to pigeons. GR205171 ([(2S-cis)-N-((2-methoxy-5(5 (trifluoromethyl)-1H-tetrazol-1-yl)-phenyl) methyl)-2-phenyl-3-piperidinamine dihydrochloride]) administered intramuscularly (1-10 mg/kg) reduced significantly the number of emetic response to cisplatin: this reduction was 60-81% (P < 0.05) for early emesis and 48-64% (P < 0.05) for the delayed response. Intracerebroventricularly administered GR205171 (30 microg/kg) also reduced the number of emetic responses: 53% (P < 0.05) in early emesis and 88% (P < 0.05) in the delayed response. However, the latency time to the first emesis was not affected by GR205171. Direct injection of cisplatin (10 microg/kg) into the fourth ventricle produced emesis, which was reduced by GR205171 administered via the peripheral or central route. Substance P-immunoreactive fibres were distributed throughout the dorsal vagal complex. These results suggest that the antiemetic effect of GR205171 on both emetic responses to cisplatin acts on a central site, and that the onset of the emetic response may be mediated partly via GR205171-insensitive mechanisms. PMID- 12586216 TI - 5-HT7 receptor-mediated relaxation of the oviduct in nonpregnant proestrus pigs. AB - The effects of 5-hydroxytryptamine (5-HT) on the muscle tonus of the ampulla and isthmus of the oviduct isolated from nonpregnant proestrus pigs were investigated, and the 5-HT receptor subtype and mechanisms of the responses were analyzed. 5-HT (1 nM-10 microM) caused a relaxation of longitudinal and circular muscles of the isthmus in a concentration-dependent manner. Tetrodotoxin did not change the relaxation, indicating a direct action of 5-HT on smooth muscle cells. The EC(50) value in the longitudinal muscle was significantly lower than that in the circular muscle but the maximum relaxations were similar. 5-HT also caused a relaxation of both muscle layers in the ampulla but the maximum relaxation of both muscles was smaller than that of the isthmus. 5-Carboxamidotryptamine (5 CT), 5-methoxytryptamine (5-MeOT) and (+/-)-8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) mimicked the relaxation of the isthmic longitudinal muscle by 5-HT, and the ranking order was 5-CT>5-HT>5-MeOT>8-OH DPAT. On the other hand, oxymethazoline, 2-methyl-5-hydroxytryptamine (2-methyl-5 HT), alpha-methyl-5-hydroxytryptamine (alpha-methyl-5-HT), [endo-N-8-methyl-8 azabicyclo-(3,2,1) oct-3-yl]-2,3-dihydro-3-ethyl-2-oxo-1H-benzimidazol-1 carboxamide (BIMU-1), ergotamine and dihydroergotamine were less effective. The relaxation by 5-HT was not decreased by ketanserin, 2-methoxy-4-amino-5 chlorobenzoic acid 2-(diethylamino)ethyl ester (tropisetron) or [1[2 (methylsulphonyl) amino ethyl]-4-piperidinyl]methyl-1-methyl-1H-indole-3 carboxylate (GR113808) but was antagonized by the following compounds in a competitive manner (with pK(b) values in parentheses): 2a-[4-(4-phenyl-1,2,3,6 tetrahydropyridyl)butyl]-2a,3,4,5-tetrahydro-benzo[cd]indol-2(1H)-one (DR4004, 9.31), methiothepin (8.91), methysergide (7.95), metergoline (7.98), mianserin (7.69), mesulergine (8.4), spiperone (6.86) and clozapine (7.4). The correlation of these pK(b) values with pK(i) values of cloned 5-HT(7) receptor or pA(2) values of porcine uterus was high and significant. 4-(3-Butoxy-4-methoxybenzyl) imidazolidin-2-one (Ro20-1724) significantly enhanced the relaxation by 5-HT but zaprinast, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) and L-nitroarginine methylester (L-NAME) did not change the responses to 5-HT. 5-HT increased cyclic AMP in the isthmic oviduct. Ampulla and isthmus contained a single class of [3H]5 CT binding sites with a similar K(d) value (0.4 nM), but the density of the receptors in the isthmus was 2.4 times higher than that in the ampulla. A significant correlation was found between the pK(i) values in the oviduct and those of the cloned 5-HT(7) receptors. Isoprenaline, sodium nitroprusside, vasoactive intestinal peptide and pituitary adenylate cyclase activating peptide were less effective in causing the relaxation of the oviduct. In conclusion, the 5-HT receptor, functionally correlated to the 5-HT(7) type, mediates the relaxation of the porcine oviduct by 5-HT through an increase in intracellular cyclic AMP. The degrees of 5-HT-induced relaxation in the isthmus and ampulla of the oviduct were different due to the heterogeneous distribution of 5-HT(7) receptors. The strongest relaxation through 5-HT(7) receptor activation suggests that 5-HT plays an important physiological role in the regulation of porcine oviduct contractility. PMID- 12586217 TI - Is height related to longevity? AB - Over the last 100 years, studies have provided mixed results on the mortality and health of tall and short people. However, during the last 30 years, several researchers have found a negative correlation between greater height and longevity based on relatively homogeneous deceased population samples. Findings based on millions of deaths suggest that shorter, smaller bodies have lower death rates and fewer diet-related chronic diseases, especially past middle age. Shorter people also appear to have longer average lifespans. The authors suggest that the differences in longevity between the sexes is due to their height differences because men average about 8.0% taller than women and have a 7.9% lower life expectancy at birth. Animal experiments also show that smaller animals within the same species generally live longer. The relation between height and health has become more important in recent years because rapid developments in genetic engineering will offer parents the opportunity to increase the heights of their children in the near future. The authors contend that we should not be swept along into a new world of increasingly taller generations without careful consideration of the impact of a worldwide population of taller and heavier people. PMID- 12586218 TI - Transforming growth factor alpha-mediated gastroprotection against stress ulceration in the rat: involvement of capsaicin-sensitive sensory neurons. AB - Exogenously administered TGF alpha has been shown to protect rodent gastric mucosa against injury caused by acid-dependent and acid-independent injury. The present study examined whether the gastroprotective effects of TGF alpha on stress-induced gastric ulceration in the rat involves activation of capsaicin sensitive sensory neurons. Fasted male SD rats were subjected to water restraint stress (WRS) for four hours. Thereafter, rats were euthanized; the stomach opened and macroscopic areas of gastric ulceration quantitated (mm(2)). Gastric tissue contents of TGF alpha and the sensory neuropeptide, calcitonin gene-related peptide (CGRP) were determined by radioimmunoassay. Prior to stress rats received TGF alpha 50, 100 or 200 microg/kg by intraperitoneal injection. Sensory denervation was accomplished by high dose capsaicin treatment. WRS caused severe ulceration in the gastric corpus; 46.1 + 6.6 mm(2). Parenteral administration of TGF alpha caused dose-dependent reduction in gastric injury: 34.7 + 4.9 mm(2) with 50 microg/kg (p < 0.05); 25.4 + 3.6 mm(2) with 100 microg/kg (p < 0.001) and 9.4 + 0.8 mm(2) with 200 microg/kg (p < 0.001). The gastroprotective action of TGF alpha (200 microg/kg, i.p.) was abolished by capsaicin-induced sensory denervation. In addition, WRS ulceration was associated with significant reduction in gastric CGRP (-42%) and TGF alpha (-48%) content. Reduction in CGRP content was prevented by TGF alpha pretreatment. We conclude that: 1) TGF alpha caused dose-dependent gastroprotection against WRS ulceration, 2) TGF alpha mediated gastric mucosal protection was prevented by capsaicin-induced sensory denervation and, 3) stress-induced injury was associated with significant reduction in gastric content of both TGF alpha and CGRP. PMID- 12586219 TI - Upregulation of beta 1-adrenergic receptors in ovariectomized rat hearts. AB - Changes in cardiac myofilament Ca(2+) activation have been demonstrated in ovariectomized rats. The underlying mechanisms responsible for these changes, however, are unknown. Accordingly, we measured both density and binding affinity of cardiac beta(1)-adrenergic receptors in sarcolemmal preparations from 10-week ovariectomized rats, pair-fed ovariectomized rats, and sham-operated control rats. Receptor protein content was also measured by immunoblotting. Deprivation of ovarian sex hormones for 10 weeks induced a significant upregulation of beta(1)-adrenergic receptors without affecting binding affinity. The same magnitude of receptor upregulation was also detected in pair-fed ovariectomized hearts. To determine which hormone is responsible for the observed increase in beta(1)-adrenergic receptor density, various sex hormone supplemental regimens were administered to ovariectomized rats. Subcutaneous injection of estrogen (5 microg/rat), progesterone (1 mg/rat), or estrogen plus progesterone three times a week all effectively prevented the upregulation of the beta(1)-adrenoceptors. Western blot analyses using polyclonal antibody of beta(1)-adrenergic receptors revealed the same pattern of changes in the protein content of the receptors in these various groups of experimental hearts as those obtained from the receptor binding assay. These results suggest a possible direct suppressive effect of ovarian sex hormones on the expression of cardiac beta(1)-adrenergic receptors. PMID- 12586220 TI - Acute ethanol administration profoundly alters poly I:C-induced cytokine expression in mice by a mechanism that is not dependent on corticosterone. AB - Polyinosinic polycytidylic acid (poly I:C) is an analog of double stranded RNA, which is a common replication intermediate for many viruses. It acts through a toll-like receptor (TLR3) to induce a group of cytokines that can mediate host resistance to viruses and some cancers. The effect of ethanol (EtOH) on induction of this set of cytokines has not been determined. Mice were treated with a single dose of EtOH (by gavage) at the same time as poly I:C was administered (intraperitoneally), and cytokine mRNA expression was measured by RNAse protection assay. Concentrations of IFN-alpha, IL-10, and IL-12 in the serum were measured by ELISA. A single dose of EtOH suppressed induction of mRNA for IFN alpha, IFN-beta, IFN-gamma, IL-6, IL-9, IL-12, and IL-15. The concentrations of IFN-alpha and IL-12 in the serum were also decreased. In contrast, IL-10 was minimally induced by poly I:C alone, but it was substantially induced by poly I:C plus EtOH. Dose response and time course studies demonstrated that significant alterations of IFN-alpha, IL-10, and IL-12 expression occurred at dosages as low as 4 g/kg (a dosage previously shown to produce blood EtOH concentrations of approximately 0.2%) and that alterations persisted at least 4-6 hr after administration of EtOH. The glucocorticoid synthesis inhibitor, aminoglutethimide, diminished corticosterone levels to normal, but did not block the effects of EtOH on cytokine expression. These results demonstrate that EtOH affects the expression of poly I:C-induced cytokines and that this action is not mediated by corticosterone. These results plus previously published findings are consistent with the idea that EtOH may be a generalized suppressor of toll-like receptor signaling. PMID- 12586221 TI - Role of intracellular Ca2+ in endothelium-dependent contraction and relaxation of rabbit intrapulmonary arteries. AB - We examined whether Ca(2+) mobilizers induce endothelium-dependent contraction and relaxation (EDC and EDR) in isolated rabbit intrapulmonary arteries. Ionomycin (10(-7) M) and A-23187 (10(-7) M), both Ca(2+) ionophores, and thapsigargin (10(-6) M), an endoplasmic reticulum Ca(2+)-ATPase inhibitor, caused a contraction in the non-contracted preparations, and a transient relaxation followed by a transient contraction and sustained relaxation in the precontracted preparations. Endothelium-removal abolished the contraction and transient relaxation (EDC and EDR) but not sustained relaxation (endothelium-independent relaxation, EIR). In the noncontracted preparations, ionomycin-induced EDC was significantly attenuated by quinacrine (10(-5) M), manoalide (10(-6) M), both phospholipase A(2) inhibitors, indomethacin (10(-5) M) and aspirin (10(-4) M), both COX inhibitors, and ozagrel (10(-5) M), a TXA(2) synthetase inhibitor. In the precontracted arteries, EDR was markedly reduced by L-NAME (10(-4) M), a NOS inhibitor, and methylene blue (10(-6) M), a guanylate cyclase inhibitor, and was enhanced by indomethacin, aspirin and ozagrel, probably due to inhibition of EDC. ZM230487, a 5-lipoxygenase inhibitor, had no effect on EDR. EIR was not affected by L-NAME, indomethacin or ZM230487. Arachidonic acid (10(-6) M) evoked EDC sensitive to indomethacin and ozagrel. L-Arginine (10(-3) M) caused EDR sensitive to L-NAME in the ionomycin-stimulated preparations. In conclusion, Ca(2+) mobilizers cause EDC and EDR via production of TXA(2) and NO, respectively. PMID- 12586222 TI - Inhibition of tumor growth in vitro by the extract of fagopyrum cymosum (fago-c). AB - Fagopyrum cymosum (Trev.) Meisn has long been used in China to treat various ailments of the lung, including lung tumors. This study investigated whether Fagopyrum cymosum extract (Fago-c) has effects on other organs. Human cancer cells derived from 10 different organs were employed, and their growths as affected by Fago-c were investigated. It was found that the growth of cancer cells from lung, liver, colon, leukocytes and bone is inhibited by Fago-c. However, cancer cells derived from prostate, cervix, ovary and brain are not sensitive to Fago-c, and the extract stimulates the growth of cancer cells from breast (MCF-7). Synergistic inhibition effect of Fago-c and daunomycin was observed in human lung cancer cells (H460). Cellular proteins from H460 cells treated with Fago-c were analyzed by 2D-gel electrophoresis. A protein (M.W./pI = 20K/5.9) was induced. The Fago-c extract was analyzed by High Performance Liquid Chromatography (HPLC). Four major and twenty minor components were identified. These studies indicate that the effect of Fago-c in inhibiting the growth of cell lines derived from certain organs. PMID- 12586223 TI - Contribution of endocannabinoids in the endothelial protection afforded by ischemic preconditioning in the isolated rat heart. AB - The aim of the present study was to assess the contribution of endogenous cannabinoids in the protective effect of ischemic preconditioning on the endothelial function in coronary arteries of the rat. Isolated rat hearts were exposed to a 30-min low flow ischemia (1 ml/min) followed by 20-min reperfusion, after which the response to the endothelium-dependent vasodilator, serotonine (5 HT), was compared with that of the endothelium-independent vasodilator, sodium nitroprusside (SNP). In untreated hearts, ischemia-reperfusion diminished selectively 5-HT-induced vasodilatation, compared with time-matched sham hearts, the vasodilatation to SNP being unaffected. A 5-min zero-flow preconditioning ischemia in untreated hearts preserved the vasodilatation produced by 5-HT. Blockade of either CB(1)-receptors with SR141716A or CB(2)-receptors with SR144528 abolished the protective effect of preconditioning on the 5-HT vasodilatation. Perfusion with either palmitoylethanolamide or 2 arachidonoylglycerol 15 min before and throughout the ischemia mimicked preconditioning inasmuch as it protected the endothelium in a similar fashion. This protection was blocked by SR144528 in both cases, whereas SR141716A only blocked the effect of PEA. The presence of CB(1) and CB(2)-receptors in isolated rat hearts was confirmed by Western blots. In conclusion, the data suggest that endogenous cannabinoids contribute to the endothelial protective effect of ischemic preconditioning in rat coronary arteries. PMID- 12586224 TI - Acetylcholine affects rat liver metabolism via type 3 muscarinic receptors in hepatocytes. AB - Although the role of acetylcholine (Ach) in hepatic glucose metabolism is well elucidated, it is still unclear if it influences gluconeogenesis, glycogenolysis and high-energy phosphate metabolism, and if it does what the mechanisms of this influence are. Therefore, using isolated perfused rat liver as a model, we have studied the effect of Ach on oxygen consumption, synthesis of glucose from lactate and pyruvate, glycogen formation, mitochondrial oxidative phosphorylation and ATP-synthesis. We have established that effects of Ach on oxygen consumption depend on its concentration. When used at a concentration of 10(-7) M, Ach exerts maximum stimulatory effect, while its infusion at 10(-6) M causes a decrease of oxygen consumption by the liver. Moreover, when used at a concentration of 10(-6) M or 10(-7) M, Ach increases rates of glucose production from the gluconeogenic substrates lactate and pyruvate, leading to enhanced glycogen content in perfused liver. It was also shown that Ach possesses a stimulating effect on alanine and aspartate aminotransferases. As detected by 31P NMR spectroscopy, continuous liver perfusion with pyruvate and lactate in the presence of Ach leads to a significant decrease of ATP level, implying enhanced energy requirements for gluconeogenesis under these conditions. Elimination of the described effects of Ach by atropine, the antagonist of muscarinic receptors, and identification of the type 3 muscarinic receptors (m3) in isolated hepatocytes as well as in whole liver, imply that Ach may exert its effect on liver metabolism through m3 receptors. PMID- 12586225 TI - New morphinan derivatives with negligible psychotropic effects attenuate convulsions induced by maximal electroshock in mice. AB - Interest in dextromethorphan (DM) has been renewed because of its anticonvulsant and neuroprotective properties. However, DM at supra-antitussive doses can produce psychotomimetic effects in humans. Recently, we demonstrated that DM exerts psychotropic effects in mice [Neurosci. Lett. 288 (2000) 76, Life Sci. 69 (2001) 615]. We synthesized a series of compounds with a modified morphinan ring system, with the intention of developing compounds that retain the anticonvulsant activity with weak psychotropic effects [Bioorg. Med. Chem. Lett. 11 (2001) 1651]. In order to extend our understanding of the pharmacological intervention of these morphinans, we assessed their behavioral effects, and then examined whether they exert protective effects on maximal electroshock convulsions (MES) in mice. Repeated treatment (20 or 40 mg/kg, i.p./day x 7) with DM or dextrorphan (a major metabolite of DM; DX) significantly enhanced locomotor activity in a dose-related manner. This locomotor stimulation was accentuated more in the animals treated with DX, and might be comparable to that of phencyclidine (PCP). By contrast, treatment with a metabolite of DM [3-methoxymorphinan (3MM) or 3 hydroxymorphinan (3HM)], 3-allyloxy-17-methylmorphinan (CPK-5), or 3 cyclopropylmethoxy-17-methylmorphinan (CPK-6) did not significantly alter locomotor activity or patterns. The behavioral effects mediated by these morphinans and PCP paralleled the effects of conditioned place preference. DM, DX, CPK-5, and CPK-6 had anticonvulsant effects against MES, while 3MM and 3HM did not show any anticonvulsant effects. We found that DM, DX, CPK-5 and CPK-6 were high-affinity ligands at sigma(1) receptors, while they all had low affinity at sigma(2) receptors. DX had relatively higher affinity for the PCP sites than DM. By contrast, CPK-5 and CPK-6 had very low affinities for PCP sites, suggesting that PCP sites are not requisites for their anticonvulsant actions. Our results suggest that the new morphinan analogs are promising anticonvulsants that are devoid of PCP-like behavioral side effects, and their anticonvulsant actions may be, in part, mediated via sigma(1) receptors. PMID- 12586226 TI - Intramuscular vascular endothelial growth factor gene therapy in patients with chronic critical leg ischemia. AB - We sought to investigate the safety and efficacy of intramuscular gene therapy with vascular endothelial growth factor (VEGF) in patients with chronic critical leg ischemia.Gene transfer was performed in 24 limbs of 21 patients with rest pain, some of whom also had nonhealing ischemic ulcers (n = 16) due to occlusive peripheral arterial disease. Between 400 microg and 2000 microg of phVEGF(165) (400 microg, n = 2; 800 microg, n = 4; 1200 microg, n = 4; 1600 microg, n = 6; and 2000 microg, n = 8) was injected directly into the muscles of the ischemic limb; the same dose was injected 4 weeks later. The ratio of blood pressures at the ankle and brachial artery was measured before and after treatment. Mean (+/- SD) plasma levels of VEGF increased significantly from 26 +/- 31 pg/mL to 63 +/- 56 pg/mL (P <0.005), and the ankle-brachial index improved significantly from 0.58 +/- 0.24 to 0.72 +/- 0.28 (P <0.001). Magnetic resonance angiography showed qualitative evidence of improved distal flow in 19 limbs (79%). Ischemic ulcers healed or improved markedly in 12 limbs (75%). Rest pain was relieved or improved markedly in 20 limbs (83%). Amputation was performed in two limbs because of wound infection. Complications were limited to transient leg edema in six limbs. Intramuscular gene therapy with VEGF(165) for patients with chronic critical leg ischemia is safe, feasible, and effective. PMID- 12586227 TI - Total hip replacement due to osteoarthritis: the importance of age, obesity, and other modifiable risk factors. AB - We studied whether several modifiable factors were associated with the risk of total hip replacement due to hip osteoarthritis among women.We identified 568 women from the Nurses' Health Study who reported total hip replacement due to primary hip osteoarthritis on questionnaires from 1990 to 1996, using a validated algorithm. The relation of potential risk factors, such as age, body mass index, physical activity, smoking, alcohol intake, and hormone use, to hip replacement was assessed using pooled logistic regression models. Higher body mass index was associated with an increased risk of hip replacement due to osteoarthritis (P for trend = 0.0001). Compared with women in the lowest category of body mass index (<22 kg/m(2)), those in the highest category of body mass index (> or =35 kg/m(2)) had a twofold increased risk (95% confidence interval [CI]: 1.4 to 2.8), whereas those in the highest category of body mass index at age 18 years had more than a fivefold increased risk (95% CI: 2.5 to 10.7). Age also had a positive association; women aged > or =70 years were nine times more likely to have hip replacement than those aged <55 years (95% CI: 5.4 to 13.9). Recreational physical activity, smoking, alcohol use, and postmenopausal hormone use were not associated with an increased risk of hip replacement. In the Nurses' Health Study, higher body mass index and older age significantly increased the risk of total hip replacement due to osteoarthritis. Part of this risk appeared to be established early in life. PMID- 12586228 TI - Value of the electrocardiogram in predicting left ventricular enlargement and dysfunction after myocardial infarction. AB - To identify electrocardiographic predictors of left ventricular enlargement or persistent dysfunction following a myocardial infarction. Baseline and predischarge 12-lead electrocardiograms (ECGs) from 272 patients with anterior myocardial infarction who were enrolled in the Healing and Early Afterload Reducing Therapy trial were evaluated and related to echocardiographic data obtained at baseline and day 90. ST-segment elevation, QRS score, and number of negative T waves were assessed at both time points. The majority of patients (87%; n = 237) received reperfusion therapy. Multivariate models were used to adjust for potential confounders, including maximal creatine kinase level and ejection fraction at baseline. None of the baseline electrocardiographic variables independently predicted ventricular enlargement or recovery of function. In contrast, the sum of ST- and maximum ST-segment elevation, and the number of leads with ST-segment elevation > or =1 mm in the predischarge ECG, were independent predictors of ventricular enlargement from baseline to day 90. Each lead with ST-segment elevation > or =1 mm was associated with 3.5 mL of ventricular enlargement (95% confidence interval [CI]: 1.6 to 5.5 mL; P <0.0001). Similarly, the sum of ST-segment elevation (odds ratio [OR] = 0.78; 95% CI: 0.69 to 0.89; P <0.0001), the maximum ST-segment elevation (OR = 0.25; 95% CI: 0.13 to 0.45; P <0.0001), and the number of leads with ST-segment elevation > or =1 mm (OR = 0.58; 95% CI: 0.45 to 0.74; P <0.0001) were independently associated with a lower likelihood of recovery of function at day 90. Predischarge ECG may be a useful tool for early identification of patients at risk of ventricular enlargement and persistent dysfunction following myocardial infarction. PMID- 12586229 TI - An invasive strategy is associated with decreased mortality in patients with unstable angina and non-ST-elevation myocardial infarction: GUSTO IIb trial. AB - There has been much debate concerning an invasive versus a conservative strategy for patients with acute coronary syndromes. The purpose of this study was to determine whether early in-hospital catheterization reduced mortality in patients with unstable angina and non-ST-elevation myocardial infarction. We performed a retrospective analysis of data collected in the Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO) IIb trial, which compared hirudin and heparin in patients with acute coronary syndromes. We identified 8011 patients with non ST-segment elevation myocardial infarction and unstable angina who were enrolled in the trial. The primary endpoints were all-cause mortality at 30 days and 1 year. Data were analyzed with multivariate hazards models and propensity scores.After accounting for inception time bias, there were 7897 patients identified, of whom 4536 patients (57%) underwent invasive therapy and 3361 (43%) underwent conservative therapy. Adjusting for propensity scores, the adjusted 30 day mortality for the invasive group was 2.5% compared with 2.7% in the conservative group (P = 0.92); at 1 year, the invasive group had a 6.2% mortality, versus 8.6% in the conservative group (P = 0.005). In a multivariate analysis that adjusted for other clinical factors, an invasive strategy was associated with lower 1-year mortality (hazard ratio = 0.46; 95% confidence interval: 0.10 to 0.84). In patients presenting with acute coronary syndromes, an invasive strategy is associated with improved survival at 1 year even after adjusting for baseline differences. PMID- 12586230 TI - The prognostic importance of anemia in patients with heart failure. AB - Physiologic studies have suggested that anemia could adversely affect the cardiovascular condition of patients with heart failure. Yet, the prognostic importance of this treatable condition is not well established by epidemiologic studies. We sought to determine the prognostic value of hematocrit level in a cohort of elderly patients hospitalized with heart failure. We studied a consecutive sample of 2281 patients aged 65 years or older who had been admitted with a principal discharge diagnosis of heart failure. Multivariate Cox proportional hazards regression was conducted to test whether hematocrit level was an independent predictor of 1-year mortality and of hospital readmission. The mean (+/- SD) age of the patients was 79 +/- 8 years; 58% (n = 1324) were women. Their median hematocrit was 38% (25th to 75th percentile, 33% to 42%). Lower hematocrits were associated with a higher mortality. After adjusting for demographic and clinical factors, each 1% lower hematocrit was associated with a 2% greater 1-year mortality (P = 0.007). Compared with patients with a hematocrit >42%, those with a hematocrit < or =27% had a 40% greater 1-year mortality (hazard ratio [HR] = 1.40; 95% confidence interval [CI]: 1.02 to 1.92; P = 0.04). This increased risk was similar to that conferred by traditional risk factors, including a left ventricular ejection fraction < or =20% (HR = 1.50; 95% CI: 1.20 to 1.86). Lower hematocrits were also associated with a greater risk of hospital readmission. Anemia is associated with an increased risk of death and rehospitalization in older patients with heart failure. Whether anemia is a direct cause of worse outcomes, or a marker for other causal factors, is not known. PMID- 12586231 TI - Effect of false-positive mammograms on return for subsequent screening mammography. AB - There has been concern that false-positive mammograms may deter women from future screening mammograms. We sought to determine whether false-positive mammograms affected follow-up for rescreening. We studied a cohort of 41,844 women in the Vermont Mammography Registry. We measured the proportion of women returning for the next screening mammogram for 30 months following an initial screening mammogram, and compared rates of follow-up screening at 18 and 30 months in women with false-positive and true-negative initial mammograms. We adjusted for potential confounders using multivariable logistic regression models. Of the 2469 women aged 50 years or older with false-positive mammograms, 67.2% (n = 1660) returned at 18 months for rescreening, compared with 63.9% (16,948/26,521) of the women with true-negative mammograms (P = 0.001). Similarly, 86.8% (2143/2469) of the women with false-positive mammograms returned at 30 months for rescreening, compared with 84.7% (22,466/26,521) of the women with true-negative mammograms (P = 0.005). After adjusting for age, use of hormone replacement therapy, prior mammography, prior false-positive mammography, and education, women with false positive mammograms were more likely to return at 18 months (odds ratio [OR] = 1.40; 95% confidence interval [CI]: 1.30 to 1.51) and at 30 months (OR = 1.30; 95% CI: 1.18 to 1.44). Despite previous concerns, false-positive screening mammograms did not discourage women from returning for subsequent screening mammography. However, other deleterious effects of false-positive results still warrant improvements in the accuracy of breast cancer screening. PMID- 12586232 TI - Stress echocardiography: a historical perspective. AB - Stress echocardiography is a popular cardiac imaging technique that provides similar diagnostic and prognostic accuracy as radionuclide stress perfusion imaging. Similar accuracy, however, should not be synonymous with being "clinically interchangeable." Because of economic cost, environmental impact, and individual biohazard exposure, a nuclear examination should be performed only when it cannot be replaced by other techniques that do not employ ionizing radiation. Nuclear medical imaging procedures, of which cardiological procedures are an important part, account for 4% of the annual effective radiation dose received by the average person in the United States. At the individual level, the radiological exposure of a single nuclear cardiology procedure conveys a low, but measurable, risk of fatal cancer, which varies from 1 in 1000 to 1 in 10,000, depending on the type of examination (higher for thallium scans; lower for technetium 99m methoxyisobutylisonitrile scans). Because the information provided by stress echocardiography and stress radionuclide perfusion imaging are similar, the choice of test should be made in the context of the environmental, biological, and economic effects of that choice. PMID- 12586233 TI - Contemporary cardiology and hysteric nucleophobia. PMID- 12586235 TI - Inhaled or systemic corticosteroids and the risk of hospitalization for hip fracture among elderly women. PMID- 12586234 TI - Prescription of QT-prolonging drugs in a cohort of about 5 million outpatients. AB - Many drugs prolong the QT interval and increase the risk of torsade de pointes. Concurrent use of two or more of these drugs can further increase the risk, but the prevalence of concurrent prescription of QT-prolonging drugs is not known. Using the administrative claims database of a national pharmaceutical benefit manager, we conducted a retrospective cohort study in 4,825,345 subjects aged 18 years or older. After identifying 50 drugs with QT-prolonging potential, and an additional 26 drugs that inhibit the metabolic clearance of QT-prolonging drugs, we measured the frequency of overlapping prescriptions for two or more of these drugs in the outpatient setting in 1999. Nearly 1.1 million subjects (22.8%) filled 4.4 million prescriptions for QT-prolonging drugs. Of these, 103,119 subjects (9.4%) filled overlapping prescriptions for two or more of the drugs or for a QT-prolonging drug and another drug that inhibits its clearance; 7249 subjects (0.7%) filled overlapping prescriptions for three or more of these drugs. Twenty-two percent of subjects who filled overlapping prescriptions were aged 65 or older; 74% were women. Antidepressants were involved in nearly 50% of the cases. Concurrent prescription of QT-prolonging drugs is common in the outpatient setting, and antidepressants are involved in half of these cases. Large pharmaceutical claims databases are useful for detecting potentially harmful prescribing behaviors, but better clinical evidence on medication safety is needed before such a system can be implemented fully. PMID- 12586236 TI - Sjogren's syndrome-associated myelopathy: response to immunosuppressive treatment. PMID- 12586237 TI - A founder effect in the hyperimmunoglobulinemia D and periodic fever syndrome. PMID- 12586238 TI - Cases from the Osler Medical Service at Johns Hopkins University. AB - A 37-year-old woman presented with increasing abdominal pain and jaundice. Six weeks before admission, she developed persistent diarrhea and jaundice of the skin. She also bruised easily, and her gums bled. In the subsequent weeks, her appetite decreased, she was fatigued, and she had nausea, vomiting, and abdominal distension. She had a history of drinking 1 quart of vodka every day for 20 years, with brief periods of abstinence; she stopped consuming alcohol 11 days before admission because it no longer provided symptomatic relief. Her past medical history was also notable for depression, including a suicide attempt 4 years earlier. She did not smoke, use illicit drugs, or have unprotected sexual intercourse. She had received no blood transfusions and had not traveled recently. She took no medications, except for occasional ibuprofen. On physical examination, she was thin and deeply jaundiced, and she trembled and responded slowly to questions. She was afebrile but tachypneic, and she had orthostatic hypotension. Her HEENT examination was notable for scleral and sublingual icterus, as well as crusted blood on her gums and teeth. The jugular veins were flat. The cardiac examination revealed tachycardia (heart rate, 103 beats per minute) without murmurs, rubs, or gallops. The abdomen was nontender and protuberant, with hypoactive bowel sounds; the spleen was not palpable, and there was no fluid wave or caput medusae. The liver percussed to 18 cm, with a smooth edge extending 10 cm below the costal margin. She had cutaneous telangiectases on her chest and bilateral palmar erythema. There was no peripheral edema. The neurologic examination was notable for asterixis. Her stool was guaiac positive. Laboratory studies revealed the following values: hematocrit, 21.2%; white blood cells, 17,310/mm(3); ammonia, 42 micromol/L; serum creatinine, 3.9 mg/dL; serum urea nitrogen, 70 mg/dL; albumin, 2.1 g/dL; total bilirubin, 26.8 mg/dL; alanine aminotransferase, 14 U/L; aspartate aminotransferase, 77 U/L; alkaline phosphatase, 138 U/L; prothrombin time, 103 seconds (international normalized ratio, 10.6); and urinary sodium, <5 mg/dL. Urinalysis revealed an elevated specific gravity and numerous muddy granular casts. Hepatitis A, B, and C serologies were negative. On abdominal ultrasound examination, there was no ascites, and the liver was echogenic. The portal and hepatic veins were patent, and the hepatic arteries were normal. The spleen measured 14 cm. What is the diagnosis? PMID- 12586239 TI - Intramuscular vascular endothelial growth factor gene therapy: fact or fiction? PMID- 12586240 TI - Obesity and hip osteoarthritis: the weight of the evidence is increasing. PMID- 12586241 TI - Rhabdomyolysis as a consequence of adrenal insufficiency. PMID- 12586242 TI - Acute abdomen due to spontaneous organ rupture in systemic lupus erythematosus. PMID- 12586243 TI - Interferon-induced hypopituitarism. PMID- 12586244 TI - Linear IgA bullous dermatosis induced by angiotensin receptor antagonists. PMID- 12586245 TI - Leptospirosis and the antiphospholipid syndrome. PMID- 12586246 TI - Acute respiratory failure, lactic acidosis, and shock associated with a compressive isolated right pleural effusion following ovarian hyperstimulation syndrome. PMID- 12586247 TI - Colchicine-induced rhabdomyolysis. PMID- 12586248 TI - Medicine as a profession--back to basics: preserving the physician-patient relationship in a challenging medical marketplace. PMID- 12586249 TI - Effects of acute myocardial ischemia on QT dispersion by dipyridamole stress echocardiography. AB - Increased dispersion of the QT interval has been observed during pacing or exercise stress testing in patients with coronary artery disease (CAD). It has not been established whether this phenomenon is a consequence of ischemia. Therefore, we sought to evaluate whether dipyridamole-induced myocardial ischemia, as directly detected by echocardiographic monitoring of regional contractile function, would affect QT dispersion. Twenty-four patients with nonsignificant and 34 patients with significant CAD but no previous myocardial infarction underwent dipyridamole stress echocardiography while not taking medications. QT dispersion was measured on a 12-lead electrocardiogram at baseline and at various times after dipyridamole infusion. Dipyridamole infusion did not influence QT dispersion in patients without CAD. QT dispersion was similarly unaffected in patients with CAD in whom dipyridamole did not induce wall motion abnormalities. In contrast, in patients with positive dipyridamole stress test findings, QT dispersion increased from 60 +/- 17 ms at baseline to 94 +/- 25 ms during peak infusion (p <0.0001), with a time course mirroring that of development of contractile abnormalities. QT dispersion returned to 63 +/- 25 ms upon relief of ischemia by administration of aminophylline. The increase in QT dispersion was significantly related to the extent of contractile dysfunction induced by dipyridamole. Although ST-segment depression occurred in only 40% of patients with positive dipyridamole stress test findings, 88% of such patients had an increase in QT dispersion. Analysis of the receiver-operating characteristic curve showed that a QT dispersion increase of > or =20 ms identified positive findings for dipyridamole stress echocardiography with 68% sensitivity and 91% specificity. Thus, QT dispersion is acutely affected by myocardial ischemia induced by the administration of dipyridamole. Measurement of QT dispersion may improve detection of stress-induced ischemia on surface electrocardiograms. PMID- 12586250 TI - Usefulness of quantitative echocardiographic techniques to predict recovery of regional and global left ventricular function after acute myocardial infarction. AB - The left ventricular response to dobutamine may be quantified using tissue Doppler measurement of myocardial velocity or displacement or 3-dimensional echocardiography to measure ventricular volume and ejection fraction. This study sought to explore the accuracy of these methods for predicting segmental and global responses to therapy. Standard dobutamine and 3-dimensional echocardiography were performed in 92 consecutive patients with abnormal left ventricular function at rest. Recovery of function was defined by comparison with follow-up echocardiography at rest 5 months later. Segments that showed improved regional function at follow-up showed a higher increment in peak tissue Doppler velocity with dobutamine therapy than in nonviable segments (1.2 +/- 0.4 vs 0.3 +/- 0.2 cm/s, p = 0.001). Similarly, patients who showed a >5% improvement of ejection fraction at follow-up showed a greater displacement response to dobutamine (6.9 +/- 3.2 vs 2.1 +/- 2.3 mm, p = 0.001), as well as a higher rate of ejection fraction response to dobutamine (9 +/- 3% vs 2 +/- 2%, p = 0.001). The optimal cutoff values for predicting subsequent recovery of function at rest were an increment of peak velocity >1 cm/s, >5 mm of displacement, and a >5% improvement of ejection fraction with low-dose dobutamine. PMID- 12586251 TI - Incremental value of myocardial contrast echocardiography for the prediction of recovery of function in dobutamine nonresponsive myocardium early after acute myocardial infarction. AB - We hypothesized that the presence of microvascular integrity, detected by myocardial contrast echocardiography (MCE) in dobutamine nonresponsive segments, may enhance identification of recovery of function, which is a surrogate marker of myocardial viability. Accordingly, 96 patients underwent dobutamine echocardiography (DE) and intravenous MCE on the same day, 4.6 +/- 1.5 days after acute myocardial infarction (AMI). Recovery of function of akinetic segments was assessed at 3 months after AMI. Of 387 akinetic segments, 102 (26%) recovered function during follow-up. Sensitivities and specificities of MCE, DE, and the combination of DE and MCE in dobutamine nonresponsive segments were 58%, 59%, and 79%, respectively (p <0.001, compared with MCE and DE) and 76%, 84%, and 69%, respectively (p <0.05 compared with DE) for predicting recovery of function. In anterior AMI, the positive and negative predictive values of MCE, DE, and the combination of DE and MCE were 47% and 88%, 57% and 89%, and 49% and 95%, respectively. Multivariate analysis using clinical characteristics, electrocardiography, biochemical factors, MCE, and DE showed that the combination of DE and MCE in dobutamine nonresponsive segments (p <0.00001) and Q-wave AMI (p = 0.002) were the only independent predictors of recovery of function. Thus, for optimum prediction of recovery of function after AMI, a combination of DE and MCE in dobutamine nonresponsive segments may be utilized. PMID- 12586252 TI - Predicting angiographic distal embolization following percutaneous coronary intervention in patients with acute myocardial infarction. AB - The aim of this study was to investigate the relation between lesion morphology identified by intravascular ultrasound (IVUS) before intervention and angiographic distal embolization after percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI). PCI for AMI has already been established as beneficial therapy, although some complications remain unresolved. Distal embolization is 1 of the important complications of PCI. Recently, some new devices have been developed for the prevention of distal embolization. However, few studies exist that look into the relation between lesion morphology and distal embolization. IVUS was performed safely in 140 consecutive patients with AMI before coronary intervention. No patient received thrombolytic therapy. From the incidence of angiographic distal embolization, patients were divided into 2 groups--an embolization group and a nonembolization group--and clinical background, IVUS, and angiographic information were evaluated. Distal embolization was observed in 12 patients (9%). Peak creatine kinase levels (3,877 +/- 2,285 vs 2,293 +/- 1,792 IU/L, p <0.05) and the incidence of angiographic thrombus (25% vs 5%, p <0.05) and intracoronary mobile mass detected by IVUS (75% vs 16%, p <0.001) were higher for patients in the embolization group. From the multivariate logistic regression analysis, only an intracoronary mobile mass detected by IVUS emerged as a predictor of distal embolization (odds ratio 53, 95% confidence interval 2.7 to 1,040, p <0.01). Patients with an intracoronary mobile mass detected by IVUS are prone to distal embolization after PCI and larger infarction. IVUS imaging before PCI may be useful for determining which patients need a distal protection device. PMID- 12586253 TI - Incremental value of exercise technetium-99m tetrofosmin myocardial perfusion single-photon emission computed tomography for the prediction of cardiac events. AB - Technetium-99m (Tc-99m) tetrofosmin single-photon emission computed tomography (SPECT) is a useful alternative to thallium-201 scintigraphy for the assessment of myocardial perfusion. This study assessed the incremental value of exercise Tc 99m tetrofosmin SPECT for the prediction of cardiac events in patients with known or suspected coronary artery disease. Exercise Tc-99m tetrofosmin SPECT imaging was performed in 655 consecutive patients. Follow-up was successful in 648 patients (98.9%). Ten patients underwent early coronary revascularization and were excluded. End points were cardiac death, nonfatal infarction, and late (>60 days) coronary revascularization. An abnormal study was defined as the presence of fixed and/or reversible perfusion defects. A summed stress score (SSS) was derived to estimate the extent and severity of perfusion defects. An abnormal scan was detected in 344 patients (54%). During a mean follow-up period of 4 +/- 1.3 years, 56 patients (9%) died (22 cardiac deaths). Nonfatal myocardial infarction occurred in 19 patients (3%), and 89 patients (14%) underwent late coronary revascularization. An abnormal scan was an independent predictor of cardiac death (hazard ratio 3.5, confidence intervals [CI] 1.1 to 12.2) and provided incremental information over clinical and exercise test data (log likelihood -133 to -125, p <0.05). The SSS provided incremental prognostic information over clinical data as well (log-likelihood -133 to -127, p <0.05) (hazard ratio 1.23, CI 1.10 to 1.38). An abnormal scan (hazard ratio 3.3, CI 1.1 to 12.2)) and the SSS (hazard ratio 1.25, CI 1.07 to 1.45)) were powerful independent predictors of the combined end point of any cardiac event. Thus, exercise Tc-99m tetrofosmin myocardial perfusion SPECT provides information incremental to clinical data for the prediction of cardiac events in patients with known or suspected coronary artery disease. PMID- 12586254 TI - Comparison of transthoracic Doppler echocardiography to intracoronary Doppler guidewire measurements for assessment of coronary flow reserve in the left anterior descending artery for detection of restenosis after coronary angioplasty. AB - Transthoracic Doppler echocardiography (TDE) has been described as a feasible and accurate technique to noninvasively assess coronary flow reserve (CFR) in the left anterior descending artery (LAD). This study was designed to evaluate whether serial assessment of CFR in the LAD using TDE allows detection of restenosis after previously performed angioplasty. Thirty-three consecutive patients with single-vessel coronary artery disease of the LAD scheduled for angioplasty underwent assessment of coronary flow velocity at rest and during adenosine-induced hyperemia in the distal LAD using high-frequency TDE. CFR was calculated as the ratio of hyperemic to basal systolic/diastolic mean velocity. Investigations were performed before and immediately after angioplasty, and at the time of control angiography after 3 months. CFR results by TDE were compared with intracoronary Doppler guide wire measurements. Adequate pulse-wave Doppler signals to measure CFR were obtained in 30 patients (91%) using TDE. There was close correlation between echocardiographically and intracoronary derived CFR results (r = 0.80, 0.79, and 0.87 before angioplasty, early after, and at 3- month control angiography, respectively; p <0.001). Using a cut-off value of CFR < or =2.0 to identify significant coronary artery disease, TDE detected LAD restenosis with a sensitivity of 89% and specificity of 90%. Thus, high-frequency TDE is a feasible technique to noninvasively assess CFR in the LAD with results closely corresponding to invasive measurements. Defining a cut-off value of CFR < or =2.0, the technique has the potential to reliably detect LAD stenosis after coronary intervention. PMID- 12586255 TI - Efficacy and safety of ezetimibe coadministered with lovastatin in primary hypercholesterolemia. AB - This multicenter, randomized, double-blind, placebo-controlled clinical study assessed the efficacy and safety of ezetimibe administered with lovastatin in primary hypercholesterolemia. After dietary stabilization, a 2- to 12-week washout period, and a 4-week single-blind placebo lead-in period, 548 patients with low-density lipoprotein (LDL) cholesterol > or =145 mg/dl (3.75 mmol/L) and < or =250 mg/dl (6.47 mmol/L) and triglycerides < or =350 mg/dl (3.99 mmol/L) were randomized to one of the following, administered daily for 12 weeks: ezetimibe 10 mg; lovastatin 10, 20, or 40 mg; ezetimibe 10 mg plus lovastatin 10, 20, or 40 mg; or placebo. The primary efficacy variable was percentage decrease in direct LDL cholesterol from baseline to end point for pooled ezetimibe plus lovastatin versus pooled lovastatin alone. Ezetimibe plus lovastatin significantly improved concentrations of LDL cholesterol, high-density lipoprotein (HDL) cholesterol, and triglycerides compared with lovastatin alone (p <0.01). The coadministration of ezetimibe provided an incremental 14% LDL cholesterol decrease, a 5% HDL cholesterol increase, and a 10% decrease in triglycerides compared with pooled lovastatin alone. Ezetimibe plus lovastatin provided mean LDL cholesterol decreases of 33% to 45%, median triglyceride decreases of 19% to 27%, and mean HDL cholesterol increases of 8% to 9%, depending on the statin dose. The coadministration of ezetimibe 10 mg plus the starting dose of lovastatin (10 mg) provided comparable efficacy to high-dose lovastatin (40 mg) across the lipid profile (LDL cholesterol, HDL cholesterol, and triglycerides). Ezetimibe plus lovastatin was well tolerated, with a safety profile similar to both lovastatin alone and placebo. The coadministration of ezetimibe and lovastatin may offer a new treatment option in lipid management of patients with hypercholesterolemia. PMID- 12586256 TI - Effectiveness of percutaneous mechanical mitral commissurotomy using the metallic commissurotome in patients with restenosis after balloon or previous surgical commissurotomy. AB - Balloon mitral valvuloplasty has been reported to give equal or less positive results after previous commissurotomy than after a first procedure. Percutaneous mechanical mitral commissurotomy (PMMC) is a new technique that has not yet been evaluated in this subset of patients. Of 1,175 PMMC procedures (1,175 patients), 173 patients (14.7%) had previous commissurotomy; patients were older (40 vs 35 years of age, p <0.0001) and more often in atrial fibrillation (34% vs 21%, p = 0.0016) than were patients who had not undergone previous commissurotomy. The baseline transmitral gradient was lower (17 +/- 8 vs 19 +/- 8 mm Hg, p <0.002) and the echocardiographic Wilkins score was higher (8.7 +/- 1.9 vs 7.6 +/- 1.8, p <0.0001) for patients who underwent previous commissurotomy. Baseline mitral valve area was comparable between the 2 groups (0.96 +/- 0.21 vs 0.93 +/- 0.24 cm(2)). Immediate results were satisfactory, although slightly less favorable after previous commissurotomy, with a final mitral valve area of 2.01 +/- 0.30 versus 2.12 +/- 0.36 cm(2) (p <0.0001), and a residual transvalvular gradient of 5.0 +/- 3.6 versus 4.2 +/- 4.1 mm Hg (p = 0.003). The rates of procedural success (93%) and severe complications (4.7%) were comparable between the 2 groups. Thus, PMMC is an effective and safe technique for the treatment of mitral restenosis after previous commissurotomy. PMID- 12586257 TI - Prenatal diagnosis by echocardiogram and outcome of absent pulmonary valve syndrome. AB - We reported echocardiographic findings and outcomes of fetuses with absent pulmonary valve syndrome diagnosed during fetal life. Cases were identified from a prospectively acquired computerized database of 18,308 pregnancies referred to a fetal cardiology center between January 1988 and July 2000. Twenty fetuses were identified with a median gestation of 23 weeks (range 18 to 36) at presentation. In 18 cases (90%), there was an associated ventricular septal defect. Eighteen cases (90%) had branch pulmonary artery diameters above the normal range. In four cases (20%), an arterial duct was present. A chromosome 22q11 deletion was identified in 2 of 9 cases (22%) in which this deletion was sought. There were 6 terminations of pregnancy (30%), 3 intrauterine deaths (15%), 5 neonatal deaths (25%), 3 infant deaths (15%), and 3 patients who did not die (15%). Ten of the 11 "liveborn" infants required early ventilation. The outcome of absent pulmonary valve syndrome diagnosed prenatally appears poor. The high morbidity and mortality is due to both cardiac disease and associated bronchomalacia. PMID- 12586258 TI - Comparative prognostic value of systolic, diastolic, and pulse pressure. PMID- 12586259 TI - Comparison of presentation, perception, and six-month outcome between women and men > or =75 years of age with angina pectoris. PMID- 12586261 TI - Atherosclerotic plaque distribution in the left anterior descending coronary artery as assessed by intravascular ultrasound. PMID- 12586260 TI - Usefulness of pharmacologic stress echocardiography for the long-term prognostic assessment of patients with typical versus atypical chest pain. PMID- 12586262 TI - Myocardial contrast echocardiography, single-photon emission computed tomography, and regional function analysis for coronary stenosis description during vasodilator stress. PMID- 12586263 TI - Usefulness of concomitant myoglobin and troponin elevation as a biochemical marker of mortality in non-ST-segment elevation acute coronary syndromes. PMID- 12586264 TI - ST-segment resolution and late (6-month) left ventricular remodeling after acute myocardial infarction. PMID- 12586265 TI - Prediction of long-term outcome by the extent of existing ST-segment deviation in a single electrocardiographic lead shortly after thrombolysis in acute myocardial infarction. PMID- 12586266 TI - Interactions between age, outcome of acute coronary syndromes, and tirofiban therapy. PMID- 12586267 TI - Effects of combined low-density lipoprotein apheresis and aggressive statin therapy on coronary calcified plaque as measured by computed tomography. PMID- 12586268 TI - Comparison of the i-STAT handheld activated clotting time with the Hemochron activated clotting time during and after percutaneous coronary intervention. PMID- 12586269 TI - Relation of Thrombolysis in Myocardial Infarction (TIMI) frame count to coronary flow parameters. PMID- 12586270 TI - Effects of left ventricular volume and ejection on myocardial blood flow measured by oxygen-15 water positron emission tomography in coronary heart disease. PMID- 12586271 TI - Association of body mass index with outcome after percutaneous coronary intervention. PMID- 12586272 TI - Usefulness of endothelin(A) receptor antagonists for the prevention of in-stent restenosis in patients with stable angina pectoris or silent myocardial ischemia. PMID- 12586273 TI - Usefulness of percutaneous left ventricular assistance to support high-risk percutaneous coronary interventions. PMID- 12586274 TI - Effect of elevated triglyceride levels on endothelium-dependent vasodilation in patients with hypercholesterolemia. PMID- 12586276 TI - Effectiveness of prehospital synchronous direct-current cardioversion for supraventricular tachyarrhythmias causing unstable hemodynamic states. PMID- 12586275 TI - Effect of serum albumin and bilirubin on the risk of myocardial infarction (the Framingham Offspring Study). PMID- 12586277 TI - P-wave signal averaging identifies patients prone to alcohol-induced paroxysmal atrial fibrillation. PMID- 12586278 TI - Effect of short- and long-duration spaceflight on QTc intervals in healthy astronauts. PMID- 12586279 TI - Endothelial dysfunction and reduced myocardial perfusion reserve in heart failure secondary to coronary artery disease. PMID- 12586280 TI - Comparison of transthoracic and transesophageal three-dimensional echocardiography for assessment of atrial septal defect diameter in children. PMID- 12586281 TI - Normal aortic arch growth and comparison with isolated coarctation of the aorta. PMID- 12586282 TI - Usefulness of peripheral artery tonometry for determining peripheral vascular responses during exercise. PMID- 12586283 TI - Diagnostic usefulness and impact on management of transesophageal echocardiography in surgical intensive care units. PMID- 12586284 TI - Comparison of one-year outcomes...ESPRIT. Enhanced Suppression of the Platelet IIb/IIIa Receptor With Integrilin Therapy. PMID- 12586285 TI - QRS interval time-related changes and prognosis in heart failure. PMID- 12586287 TI - Disappearance of septal Q waves following percutaneous transluminal septal myocardial ablation for obstructive hypertrophic cardiomyopathy. PMID- 12586289 TI - A study of susceptibility of primary human Kupffer cells to hepatitis C virus. AB - BACKGROUND/AIMS: Histological examination of a liver-biopsy from a patient with chronic hepatitis C shows activated Kupffer cells. In vitro infection of human Kupffer cells (KC) was performed to study their interaction with hepatitis C virus (HCV). METHODS: KC, isolated by collagenase perfusion and centrifugal elutriation, were infected with various HCV positive sera. The presence of the viral genome was followed, at different times, quantitatively by a branched-DNA assay and qualitatively by reverse transcriptase-nested polymerase chain reaction. A strand-specific assay performed with the thermostable enzyme rTth was used to detect the synthesis of a negative replicative intermediate. Cytopathic effect was examined by electron microscopy. Production of cytokines and inducible nitric oxide synthase was evaluated in the supernatants. RESULTS: Quantification of HCV-RNA showed that the level of viral RNA associated with KC after adsorption decreased rapidly. Genomic viral RNA disappeared within 5 days of infection. Negative-strand RNA was never detected in any of these experiments. No cytopathic effects could be detected at any time. KC did not produce inflammatory nor antiviral cytokines. CONCLUSIONS: Our results strongly suggest that primary cultures of KC are not permissive for HCV in vitro. PMID- 12586290 TI - A comparison of fibrosis progression in chronic liver diseases. AB - BACKGROUND/AIMS: No study has compared the liver fibrosis progression rates among chronic liver diseases and the risk factors in order to better organize screening strategies. METHODS: A total of 4852 patients were retrospectively studied (chronic hepatitis C (HCV) [n=2313], human immunodeficiency virus (HIV)-HCV co infection (HIV-HCV [n=180]), hepatitis B (HBV [n=777]), alcoholic liver disease (ALD [n=701]), primary biliary cirrhosis (PBC [n=406]), genetic hemochromatosis (GH [n=383]) auto-immune hepatitis (AIH [n=57]) and delta hepatitis (n=35). The fibrosis progression rates were estimated from birth and from the date of exposure, when known, to the first biopsy. RESULTS: There were highly significant differences in the rates of fibrosis progression, the most rapid being HIV-HCV co infection (50% cirrhosis percentile at 52 years of age) and the slowest being PBC (50% cirrhosis percentile at 81 years). There was an acceleration of fibrosis progression with aging. Fibrosis progression was slower in females compared with males for HCV, HBV, GH, and PBC. In contrast, in ALD, the fibrosis progression was more rapid in females. CONCLUSIONS: Rates of fibrosis progression differ markedly between the predominant causes of chronic liver disease, and according to age and gender. Patients with HIV-HCV co-infection are at particularly high risk of fibrosis progression. PMID- 12586291 TI - Incidence and natural history of small esophageal varices in cirrhotic patients. AB - BACKGROUND/AIMS: The incidence and natural history of small esophageal varices (EV) in cirrhotics may influence the frequency of endoscopies and the decision to start a pharmacological treatment in these patients. METHODS: We prospectively evaluated 206 cirrhotics, 113 without varices and 93 with small EV, during a mean follow-up of 37+/-22 months. Patients with previous gastrointestinal bleeding or receiving any treatment for portal hypertension were excluded. Endoscopy was performed every 12 months. RESULTS: The rate of incidence of EV was 5% (95%CI: 0.8-8.2%) at 1 year and 28% (21.0-35.0%) at 3 years. The rate of EV progression was 12% (5.6-18.4%) at 1 year and 31% (21.2-40.8%) at 3 years. Post-alcoholic origin of cirrhosis, Child-Pugh's class (B or C) and the finding of red wale marks at first examination were predictors for the variceal progression. The two years risk of bleeding from EV was higher in patients with small varices upon enrollment than in those without varices: 12% (95% CI: 5.2-18.8%) vs. 2% (0.1 4.1%); (P<0.01). Predictor for bleeding was the presence of red wale marks at first endoscopy. CONCLUSIONS: In patients with no or small EV, endoscopy surveillance should be planned taking into account cause and degree of liver dysfunction. PMID- 12586292 TI - Bradykinesia in minimal hepatic encephalopathy is due to disturbances in movement initiation. AB - BACKGROUND: One of the predominant symptoms of early stages of hepatic encephalopathy (HE) is bradykinesia. AIMS: To further analyze the pathophysiology of bradykinesia in HE. METHODS: A three-dimensional computer-assisted movement analysis was performed in 36 cirrhotics with grade 0-I HE compared to 18 controls selected with regard to sex and age and 16 patients with Parkinson's disease (PD). Four types of movement were studied: finger tapping, hand tapping, pronation/supination of the forearm and flexion/extension in the hip joint. RESULTS: The patients with PD presented with a decrease of the maximal movement velocity (VMAX) and a prolongation of the time needed to reach VMAX (VTIME). In patients with minimal or grade I HE, the VMAX of all movements was unchanged compared to controls while the VTIME was significantly prolonged. This was caused by a delay before the beginning of each new part of the diadochokinetic movement cycle. CONCLUSIONS: The data suggest an impairment of movement initiation as main cause of bradykinesia in early HE. PMID- 12586293 TI - Influence of leptin in the development of hepatic fibrosis produced in mice by Schistosoma mansoni infection and by chronic carbon tetrachloride administration. AB - BACKGROUND/AIMS: Leptin, a product of the obese (ob) gene is present in activated stellate cells. This study investigated whether leptin is essential for the development of hepatic fibrosis caused by various agents. METHODS: Control and ob/ob mice were infected with Schistosoma mansoni or were administered chronic carbon tetrachloride to cause hepatic fibrosis. RESULTS: Fibrosis developed in both ob/ob and control mice. However, the amount of histologically detectable fibrosis and the increase in liver hydroxyproline content was significantly greater in both models of fibrosis for treated controls than for treated ob/ob mice. Fibrosis was associated with higher secretion of TGFbeta1 from spleen cells of treated control than treated ob/ob mice. Chronic leptin administration in ob/ob mice infected with Schistosoma mansoni resulted in an increase in the amount of fibrosis caused by Schistosoma mansoni, eliminating any significant differences in the amount of fibrosis between infected ob/ob mice and control mice. It also eliminated any significant difference in TGFbeta1 secretion between the infected ob/ob and infected control mice. CONCLUSIONS: This study shows that leptin deficiency decreases but does not eliminate hepatic fibrosis produced by Schistosoma mansoni and carbon tetrachloride administration. The effect of leptin in potentiating fibrogenesis is most likely mediated by TGFbeta1. PMID- 12586294 TI - Phyllanthus amarus has anti-inflammatory potential by inhibition of iNOS, COX-2, and cytokines via the NF-kappaB pathway. AB - BACKGROUND/AIMS: Phyllanthus amarus is a herbal medicine traditionally applied in the treatment of viral hepatitis. Aim of this study was to investigate potential anti-inflammatory properties of standardized P. amarus extracts concerning a potential influence of P. amarus on endotoxin-induced nitric oxide synthase (iNOS), cyclooxygenase (COX-2), and cytokine production in vivo and in vitro. METHODS: Investigations were performed in rat Kupffer cells (KC), in RAW264.7 macrophages, in human whole blood, and in mice. Cells were stimulated with lipopolysaccharides (LPS) in the presence or absence of P. amarus extracts (hexane, EtOH/H(2)O), mice were treated with galactosamine/LPS as a model for acute toxic hepatitis. Nitrite was measured by Griess assay, prostaglandin E(2) (PGE(2)) by radioimmunoassay, and cytokines by enzyme-linked immunosorbent assay. iNOS and COX-2 were determined by Western blot, activation of NF-kappaB and AP-1 by EMSA. RESULTS: P. amarus EtOH/H(2)O and hexane extracts showed an inhibition of LPS-induced production of NO and PGE(2) in KC and in RAW264.7. The extracts also attenuated the LPS-induced secretion of tumor necrosis factor (TNF-alpha) in RAW264.7 as well as in human whole blood. Both extracts reduced expression of iNOS and COX-2 and inhibited activation of NF-kappaB, but not of AP-1. P. amarus inhibited induction of interleukin (IL)-1beta, IL-10, and interferon-gamma in human whole blood and reduced TNF-alpha production in vivo. CONCLUSIONS: This work shows that standardized extracts of P. amarus inhibit the induction of iNOS, COX-2, and TNF-alpha. Therefore, we report for the first time an anti inflammatory potential of this traditionally employed herbal medicine both in vitro and in vivo. PMID- 12586296 TI - Rate of natural disease progression in patients with chronic hepatitis C. AB - BACKGROUND/AIMS: The interval at which liver biopsy should be repeated in untreated patients with chronic hepatitis C is not defined. We examined fibrosis change by METAVIR scoring in these patients in whom two or more liver biopsies were available. METHODS: One hundred and eighty patients with histologically proven chronic hepatitis C were studied. Mean delay between biopsies was 3.67+/ 2.69 years and 3.08+/-1.43 in the 16 patients having three biopsies. Univariate and multivariate analyses were performed to determine factors associated with liver fibrosis progression. RESULTS: Median rate of fibrosis progression per year was 0.04 (0.00-0.55) to first biopsy, 0.00 (-0.84-1.02) between first and second biopsy (NS), and 0.17 (0.00-1.50) between second and third biopsy (P<0.05). In multivariate analysis, only age at first biopsy >40 years (OR=5) (2-12) and alcohol consumption of 1-50 g per day (OR=4) (2-12) and more than 50 g per day (OR=8) (3-23) were associated with severe fibrosis. The number of patients who increased in fibrosis stage was significantly higher after 4 years (P<0.02). CONCLUSIONS: An interval of at least 4-5 years is needed between liver biopsies to measure change in patients with mild liver disease. PMID- 12586297 TI - The influence of hepatitis B virus genotype on the development of lamivudine resistance during long-term treatment. AB - BACKGROUND/AIMS: Genotype-dependent development of lamivudine resistance in hepatitis B virus (HBV) has not been reported. METHODS: We determined the cumulative rate of emergence of YMDD motif mutant in 213 Japanese patients with chronic hepatitis B infected with genotype A, B, or C and treated with lamivudine for more than 1 year. RESULTS: The emergence rate of lamivudine resistance was independent of the genotype (A, B, and C). In contrast, the emergence rate was significantly higher in the Ba ("a" stands for Asia) subgroup of HBV than in Bj ("j" for Japan) subgroup (P<0.05). For genotype C (HBV/C), the emergence rate in hepatitis B e antigen (HBeAg)-positive was significantly higher than in HBV/C HBeAg-negative (P<0.05), but the rate in HBV/B-HBeAg-positive was similar to HBV/B-HBeAg-negative. Only four patients with HBV/C developed severe acute exacerbation of hepatitis accompanied by emergence of YMDD mutant, but none of the patients with other genotypes developed such complication. CONCLUSIONS: Our results suggest that lamivudine resistance in HBV does not seem to depend on the genotype but rather on the subgroup of HBV/B. The results also suggest that lamivudine resistance according to HBeAg state might be different between HBV/B and HBV/C. Large-scale prospective studies of each genotype should be conducted in the future to confirm these findings. PMID- 12586295 TI - Positional mapping for amplified DNA sequences on 1q21-q22 in hepatocellular carcinoma indicates candidate genes over-expression. AB - BACKGROUND/AIMS: Comparative genomic hybridization analysis on hepatocellular carcinoma (HCC) indicated frequent gains of 1q and an amplicon at 1q21-q22. Current cytogenetic evidences confer much importance on 1q21-q22, where a role in drug resistance, tumor metastasis and shorter patient survival had been implicated. METHODS: Using positional mapping by interphase cytogenetics, we investigated the amplicon 1q21-q22 in five HCC cases. Three amplification maxima represented by yeast artificial chromosomes (YACs) 955E11, 876B11 and 945D5 that mapped to regions 1q21.1, 1q21.2 and 1q22, respectively, were indicated. We further investigated candidate genes expression in the mapped YACs by quantitative reverse-transcription-polymerase chain reaction. A panel of genes encoding protein transcripts involved in apoptosis, cell cycle progression, calcium binding and jumping translocation was studied. RESULTS: Among ten HCC cases with the amplicon 1q21-q22 examined, we found a significant gene expression level of JTB, SHC1, CCT3 and COPA in the tumors than the paired adjacent non malignant liver tissues (P< or =0.04). CONCLUSIONS: Our interphase findings on 1q21-q22 pinpointed three affected loci between D1S305 and D1S2369. Up-regulation of candidate genes identified within these over-represented regions may represent targets in the progression of HCC and may carry prognostic significance. PMID- 12586298 TI - The effect of lamivudine therapy in hepatic decompensation during acute exacerbation of chronic hepatitis B. AB - BACKGROUND/AIMS: Severe acute exacerbation (AE) of chronic hepatitis B (CHB) can lead to hepatic decompensation and death. The aim of this study was to investigate the effect of lamivudine therapy in hepatic decompensation during such AEs. METHODS: In a 10-month period, a total of 60 consecutive AE patients with jaundice and prolonged prothrombin time over 3s were treated with lamivudine 150 mg daily. As a historical control, another 31 CHB patients with AE resulting in hepatic decompensation hospitalized in an immediate past 6-month period were enrolled for comparison. RESULTS: Patients in both groups were comparable in clinical and biochemical features. After a median treatment period of 6 weeks (range 1-48 weeks), all of the 25 patients with pretherapy bilirubin level < 20 mg/dl in the treatment group survived, while five (25%) of 20 patients in the control group died (P=0.013; odds ratios, 2.667; 95% confidence interval, 1.787 3.979). However, the mortality rate was similar in patients with pretherapy bilirubin level > or =20 mg/dl in both groups. CONCLUSIONS: These results suggest that lamivudine may prevent fatality in CHB patients with hepatic decompensation if therapy starts early enough or before serum bilirubin level rise over 20 mg/dl, but helps little if serum level already risen over that level. PMID- 12586299 TI - Antiviral effect of adefovir in combination with a DNA vaccine in the duck hepatitis B virus infection model. AB - BACKGROUND/AIMS: Combination of antiviral drugs with immunotherapeutic approaches may be a promising approach for the treatment of chronic hepatitis B. We used the duck HBV (DHBV) infection model to evaluate the efficacy of the combination of adefovir with DNA-immunization by comparison with the respective monotherapies. METHODS: Pekin ducks chronically infected with DHBV received adefovir treatment alone or in association with intramuscular immunization with a plasmid (pCI preS/S) expressing the DHBV large envelope protein. Ducks immunized with pCI preS/S plasmid alone and two control groups receiving empty plasmid injections or no treatment were followed in parallel. RESULTS: All animals treated with adefovir showed a marked drop in viremia titers during drug administration, followed by a rebound of viral replication after drug withdrawal. Eight weeks after the third DNA boost, the median of viremia within the duck group receiving the combination therapy tended to be lower compared to that of the other groups. In addition, our results suggest a trend to an additive effect of adefovir and DNA vaccine since a 51% decrease in DHBV DNA was observed in autopsy liver samples from combination therapy group, whereas pCI-preS/S or adefovir monotherapies decreased intrahepatic viral DNA by 38 and 14%, respectively. This effect was sustained since it was observed 12 weeks after the end of therapy. CONCLUSIONS: Our results suggest that combination of adefovir with DNA-vaccine may be able to induce a sustained antiviral effect in vivo. PMID- 12586300 TI - HFE mutations and chronic hepatitis C: H63D and C282Y heterozygosity are independent risk factors for liver fibrosis and cirrhosis. AB - BACKGROUND/AIMS: The impact of heterozygous HFE mutations on the course of chronic hepatitis C and iron indices was studied. METHODS: Ferritin, transferrin saturation (TS), serum iron, C282Y and H63D mutations were determined in 401 patients with chronic hepatitis C virus (HCV) infection and 295 healthy controls. Liver histologies were available in 217 and HCV genotypes in 339 patients. RESULTS: Allele frequencies of the C282Y and H63D mutation did not differ between HCV patients and healthy controls (6.95 vs. 6.2%; 14.75 vs. 16.4%; n.s.). HFE heterozygous HCV patients had higher ferritin (349+/-37 vs. 193+/-15 microg/l; P<0.0005), TS (38+/-2 vs. 32+/-1%; P<0.0005), serum iron (144+/-6 vs. 121+/-3 microg/dl; P<0.0005), semiquantitative liver iron staining (0.26+/-0.07 vs. 0.09+/-0.03; P<0.006) and fibrosis scores (1.9+/-0.2 vs. 1.4+/-0.1; P<0.003) compared to HFE wildtypes. By multivariate regression analysis odds ratios for liver cirrhosis were 5.9 (confidence interval (CI) 1.6-22.6; P<0.009) for C282Y heterozygotes and 2.9 (CI 1.0-8.4; P<0.05) for H63D heterozygotes compared to HFE wildtypes. Considering all HFE heterozygous HCV patients, odds ratios of 3.6 (CI 1.4-9.3; P<0.009) for cirrhosis and 3.1 (CI 1.3-7.3; P<0.009) for fibrosis were calculated. CONCLUSIONS: C282Y or H63D heterozygosity is an independent risk factor for liver fibrosis and cirrhosis in HCV infected individuals. Screening for HFE mutations should be considered in HCV infection. PMID- 12586301 TI - The expression of hepatitis B spliced protein (HBSP) encoded by a spliced hepatitis B virus RNA is associated with viral replication and liver fibrosis. AB - BACKGROUND/AIMS: We have previously demonstrated the in vivo expression of a new spliced hepatitis B virus (HBV) protein (HBSP) encoded by a singly spliced pregenomic RNA. The present study was designed to evaluate the impact of HBSP expression on the clinical status and liver pathology of HBV infection. METHODS: Sera from 125 chronic HBV carriers were tested for the presence of HBSP antibodies by an indirect enzyme-linked immunosorbent assay test. The severity of liver damage was evaluated using the Knodell score. RESULTS: Anti-HBSP antibody prevalence in HBV chronic carriers was 46%. We highlighted the concomitant expression of HBSP protein and anti-HBSP antibody. An association between anti HBSP antibody detection and serum markers of HBV replication was demonstrated. With respect to HBV-related liver disease, an association was only observed with the severity of fibrosis. Furthermore, an elevation of secreted tumor necrosis factor alpha (TNFalpha), but not of soluble TNFalpha receptor 75, was observed in anti-HBSP-antibody-positive patients. Multivariate analysis showed that anti-HBSP antibody detection was independently associated with viral replication, severity of fibrosis and elevated TNFalpha secretion. CONCLUSIONS: Our data suggest the hypothesis that HBSP might play a role in the natural history of HBV infection and may be involved in the pathogenesis and/or persistence of HBV infection. PMID- 12586302 TI - The presence of an intrahepatic cytotoxic T lymphocyte response is associated with low viral load in patients with chronic hepatitis C virus infection. AB - BACKGROUND/AIMS: The role of cytotoxic T lymphocytes (CTL) in limiting viral replication and producing hepatocellular injury in patients with chronic hepatitis C virus (HCV) infection is controversial. METHODS: Intrahepatic and peripheral blood HCV-specific CTL activity against the entire HCV polyprotein was assessed in 26 patients. CTL responses were assessed after effector lymphocytes were re-stimulated for 6 days in vitro using HCV-vaccinia virus-infected autologous cells expressing HCV antigens. Serum and hepatic viral loads were measured and immunohistochemistry for CD3 and CD8 was performed to localise and enumerate effector cells in liver. RESULTS: A positive CTL response was detected in 39/52 (75%) of assays conducted with intrahepatic mononuclear cells and 21/52 (40%) of peripheral blood assays (P<0.001). The presence of an intrahepatic CTL response was associated with low hepatic viral load (P=0.004). Hepatic lobular infiltration by CD8(+)T cells correlated weakly with serum alanine aminotransferase levels (r=0.42, P=0.04) and no relationship was demonstrated between CTL activity and histological evidence of liver damage. CONCLUSIONS: HCV specific CTL activity is found more commonly in liver than in blood. An inverse relationship between CTL responses and viral load supports the hypothesis that HCV-specific CTL limit viral replication in patients with chronic HCV infection. PMID- 12586303 TI - Rating fibrosis progression in chronic liver diseases. PMID- 12586304 TI - Evaluation and follow-up of patients with cirrhosis and oesophageal varices. PMID- 12586305 TI - Budd-Chiari syndrome: a review by an expert panel. PMID- 12586306 TI - A portal vein cavernoma mimicking a pancreatic mass. PMID- 12586307 TI - Fatal hepatic veno-occlusive disease associated with terbinafine in a liver transplant recipient. PMID- 12586308 TI - Parvovirus B19 and "cryptogenic" chronic hepatitis. PMID- 12586310 TI - The telomere hypothesis for progressive liver cirrhosis. PMID- 12586309 TI - Non HFE-related haemochromatosis: exclusion of the 1q21 zinc-iron regulated transporter-like (ZIRTL) gene in juvenile haemochromatosis. PMID- 12586311 TI - Growth hormone releasing factor decreases long form leptin receptor expression in porcine anterior pituitary cells. AB - Pituitary cells from six pigs, 180-200 days of age, were studied in primary culture to determine if growth hormone releasing factor (GRF) affects long form leptin receptor (Ob-Rl) expression. On Day 4 of culture, 10(5) live cells per well were challenged with either 0, 10(-6), 10(-7) or 10(-8)M [Ala15]-hGRF-(1 29)NH(2) (3 wells per treatment per pig). Secretion of growth hormone (GH) into the media and pituitary Ob-Rl mRNA expression were determined at 4 h after treatment. Media were analyzed for GH by radioimmunoassay, and total RNA was isolated from cells for determination of Ob-Rl expression by semi quantitative reverse transcription PCR. Basal GH concentration was 32+/-2 ng per 10(5) cells per well (n=18 wells) for 4 h. Relative to control at 4 h, 10(-6),10(-7) and 10( 8)M GRF increased (P<0.01) GH secretion by 151, 129 and 120%, but decreased (P<0.05) Ob-Rl expression by 32, 50 and 38%, respectively. These results indicate that GRF directly modulates Ob-Rl expression at the level of the pituitary, and thereby playing a role in regulating pituitary sensitivity to leptin. PMID- 12586312 TI - Glucagon-like peptide 2 function in domestic animals. AB - Glucagon-like peptide 2 (GLP-2) is a member of family of peptides derived from the proglucagon gene expressed in the intestines, pancreas and brain. Tissue specific posttranslational processing of proglucagon leads to GLP-2 and GLP-1 secretion from the intestine and glucagon secretion from the pancreas. GLP-2 and GLP-1 are co-secreted from the enteroendocrine L-cells located in distal intestine in response to enteral nutrient ingestion, especially carbohydrate and fat. GLP-2 secretion is mediated by direct nutrient stimulation of the L-cells and indirect action from enteroendocrine and neural inputs, including GIP, gastrin-releasing peptide (GRP) and the vagus nerve. GLP-2 is secreted as a 33 amino acid peptide and is rapidly cleaved by dipeptidylpeptidase IV (DPP-IV) to a truncated peptide which acts as a weak agonist with competitive antagonistic properties. GLP-2 acts to enhance nutrient absorption by inhibiting gastric motility and secretion and stimulating nutrient transport. GLP-2 also suppresses food intake when infused centrally. The trophic actions of GLP-2 are specific for the intestine and occur via stimulation of crypt cell proliferation and suppression of apoptosis in mucosal epithelial cells. GLP-2 reduces gut permeability, bacterial translocation and proinflammatory cytokine expression under conditions of intestinal inflammation and injury. The effects of GLP-2 are mediated by a G-protein-linked receptor that is localized to the intestinal mucosa and hypothalamus. The intestinal localization of the GLP-2R to neural and endocrine cells, but not enterocytes, suggests that its actions are mediated indirectly via a secondary signaling mechanism. The implications of GLP-2 in domestic animal production are largely unexplored. However, GLP-2 may have therapeutic application in treatment of gastrointestinal injury and diarrheal diseases that occur in developing neonatal and weanling animals. PMID- 12586313 TI - Detection and quantification of mRNA expression of alpha- and beta-adrenergic receptor subtypes in the mammary gland of dairy cows. AB - Adrenergic receptors are pharmacologically classified into the receptor types alpha(1), alpha(2), beta(1), beta(2), and beta(3). Structural differences and varying affinities in radioligand binding studies lead to a further classification of alpha(1)- and alpha(2)-receptors into subtypes which are termed alpha(1A) (formerly alpha(1C)), alpha(1B), and alpha(1D) (formerly alpha(1AD)), and alpha(2AD), alpha(2B), and alpha(2C), respectively. mRNA expression of all but one alpha-adrenergic receptor subtypes and of all beta-adrenergic receptor types was measured quantitatively in total RNA extracted from mammary tissue of 10 lactating dairy cows by real-time reverse transcription (RT) polymerase chain reaction (PCR). mRNA expression of alpha(1)-adrenergic receptors was highest for the alpha(1A)-subtype followed by alpha(1B), whereas the alpha(1D)-subtype could not be detected. The highest mRNA expression of alpha(2)-adrenergic receptors was found for the alpha(2AD)-subtype, followed by alpha(2B) and alpha(2C). Within the beta-adrenergic receptors, the beta(2)-receptor type was most highly expressed, followed by beta(1) and beta(3). In conclusion, eight of nine adrenergic receptors classified to date were detected and relatively quantified in the mammary gland of dairy cows. PMID- 12586314 TI - Thyrotropin-releasing hormone mediates serotonin-induced secretion of GH in cattle. AB - Serotonin stimulates secretion of growth hormone (GH) in cattle, but the mechanism is unknown. In rats, thyrotropin-releasing hormone (TRH) mediates serotonin-induced secretion of GH. We hypothesized that the same is true in cattle. Cattle were fed for 2h daily to synchronize secretion of GH, such that concentrations of GH were high before and low after feeding. Our first objective was to determine whether or not feeding suppresses serotonin receptor agonist (quipazine) induced secretion of GH. Holstein steers were injected with quipazine (0.2 mg/kg BW) either 1 h before or 1 h after feeding. Quipazine-induced secretion of GH which did not differ in magnitude before and after feeding. If TRH mediates serotonin-induced secretion of GH, then magnitude of TRH-induced secretion of GH should not be different before and after feeding (our second objective). Sixteen meal-fed Holstein steers were injected with 0.3 microg TRH/kg BW either 1 h before or 1 h after feeding. Indeed, magnitude of TRH-induced secretion of GH before and after feeding was not different. Our third objective was to inhibit endogenous TRH with 3,5,3'-triiodothyronine (T(3)) and examine basal, GH-releasing hormone (GHRH)-, TRH- and quipazine-induced secretion of GH. Sixteen Holstein steers were injected daily with either T(3) (3 or 6 microg/kg BW) or vehicle for 20 days and then challenged sequentially with vehicle or GHRH, TRH, or quipazine. T(3) did not affect basal, GHRH- or TRH-induced secretion of GH, but reduced basal secretion of thyroxine. T(3) reduced but did not completely block quipazine-induced secretion of GH. In conclusion, TRH mediates, in part, serotonin-induced secretion of GH in cattle. PMID- 12586315 TI - Long-term recombinant porcine somatotropin (PST) treatment mitigates the responses to subchronic lipopolysaccharide in swine. AB - The effect of multiple lipopolysaccharide (LPS) challenges in swine undergoing long-term treatment with porcine somatotropin (PST) was determined. Changes in aspartate serine transaminase (AST) occurred only at 24h following the first LPS challenge dose (P<0.05), while PST treatment moderated any change from occurring. Nonesterified free fatty acid (NEFA) levels were elevated in PST treated animals for the first 3 days following daily LPS treatment (P<0.05), while LPS treatment alone had no effect on plasma NEFA levels. Plasma urea nitrogen (PUN) levels were unchanged by LPS following the initial LPS challenge, but were decreased following the second challenge dose (P=0.014). These changes were long lasting, with a return to normal PUN levels not evident until Day 6. The PST treatment mitigated changes in PUN (P<0.05) when LPS was administered. Haptoglobin plasma levels, along with lipid peroxide production were not affected by LPS challenge or PST administration. LPS challenge reduced the levels of immunoreactive heat shock protein 70 (HSP70) throughout the entire challenge period (P<0.001). PST LPS animals had normal levels of this protein. The results of the present study demonstrate that long-term PST treatment mitigates the adverse effects of subchronic LPS administration. PMID- 12586316 TI - Matrix metalloproteinases-2 and -9 activities in bovine follicular fluid of different-sized follicles: relationship to intra-follicular inhibin and steroid concentrations. AB - Matrix metalloproteinases (MMPs) play very important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, ovulation and atresia. The aim of the present study was to determine the content of gelatinases in follicular fluid in various sized bovine follicles. Bovine ovaries were collected from local slaughterhouse and follicular fluid from follicles of 2 to over 25 mm in diameter was collected. Gelatinase activity within the follicular fluid was analyzed by gelatin zymography. The concentration of inhibin in the follicular fluid was also measured by immunoblot analysis. The proMMP-2 and alpha subunit (alphaN) inhibin was detected in all follicles regardless of their size. The abundance of proMMP-2 varied with follicular size, while alphaN inhibin increased significantly (P<0.01) in follicles of 10-14 and 15-20 mm in size. There was a positive and negative correlation between estradiol (E(2)) and progesterone (P(4)) concentrations with abundance of proMMP-2, respectively. Follicles of diameter over 25 mm had greater proMMP-9 activity than other follicles. These same follicles had significantly (P<0.01) lower inhibin levels than follicles of 10-14 and 15-20 mm in size. In conclusion, these results suggest a significant role of these proteases in growth and development of bovine follicle, particularly proMMP-2 and active MMP-2 activities in the follicular fluid could serve as markers of follicular health while abundance of proMMP-9 may possibly denote a follicular cyst. PMID- 12586317 TI - A 2-year prospective study on the effects of depot medroxyprogesterone acetate on bone mass-response to estrogen and calcium therapy in individual users. AB - The primary aim of this prospective 2-year follow-up study was to investigate the effect of depot medroxyprogesterone acetate (DMPA) on the maintenance of bone mass in women aged 30-45 years. The effects of estrogen or calcium substitution during the second year of follow-up was investigated in seven DMPA users with a high annual bone loss during the first year. The bone mass of 35 users of DMPA and 10 women without hormonal contraception was investigated using peripheral quantitative computed tomography. The baseline cortical and trabecular bone mass (TBM) and the annual change was not different in DMPA users and controls. Over 24 months we measured an increase in TBM of 0.6% and a decrease in cortical bone mass of 0.1% in exposed women. Some but not all of the DMPA users with a bone loss during the first year could be successfully treated with estradiol or calcium. In conclusion, we did not observe an accelerated bone loss in DMPA users aged 30-45 years. PMID- 12586318 TI - Acceptability of the long-term contraceptive levonorgestrel-releasing intrauterine system (Mirena): a 3-year follow-up study. AB - This study investigated the long-term acceptability of a levonorgestrel-releasing intrauterine system (LNG-IUS) in 165 women after 6 and 36 months of use. Changes in menstrual bleeding pattern were experienced by 161 (98%) women, with a cessation or transient absence of menstruation occurring in 75 (47%) and 14 (9%) women, respectively. Amenorrhea was considered by most women (81%) as a positive change. The proportion of women with menstrual pain was reduced from 60% before use to 29% after 36 months of use with the LNG-IUS. Fear of an unwanted pregnancy became less widespread with duration of use and the device had no disturbing effects on the women or their partners during sexual intercourse. The number of women expressing that they were very satisfied with the LNG-IUS was 69% and 77% after six months and 36 months of use, respectively. LNG-IUS is a well-accepted contraceptive method, without negative influences on the sexual relationship between users and their partners, and is suitable for women requiring long-term reversible contraceptive protection. PMID- 12586319 TI - Serum distribution of the major metabolites of norgestimate in relation to its pharmacological properties. AB - Norelgestromin (NGMN) and levonorgestrel (LNG) are the main active metabolites of norgestimate (NGM), but their relative contributions to the pharmacological effects of NGM are unclear. We have therefore determined the serum distribution of these NGM metabolites and assessed their steady-state concentrations in women following >or=3 cycles of oral contraceptive (OC) use. The administration of 250 microg NGM/35 microg ethinyl estradiol (EE) resulted in significantly higher sex hormone-binding globulin (SHBG) levels (p = 0.002), and 30% lower serum non protein-bound (NPB) levels of testosterone, when compared to treatment with 150 microg LNG/30 microg EE. We also confirmed that NGMN does not bind to SHBG, and found that 97.2% of this metabolite is bound to albumin while only 2.8% is in the NPB fraction. In contrast, most of the LNG was bound to SHBG (92.5% and 87.2% after NGM/EE and LNG/EE treatment, respectively), and the NPB fraction of LNG (0.7%) during NGM/EE treatment was lower (p < 0.001) than during LNG/EE treatment (1.4%). Combining these serum distributions with the C(max) and AUC(0-24h) data obtained after NGM/EE treatment gave NPB and albumin-bound values of NGMN that were much greater than the corresponding LNG values. Furthermore, the C(max) and AUC(0-24h) values for NPB LNG during NGM/EE treatment were estimated to be lower than during LNG/EE treatment. Since LNG is primarily bound by SHBG, its access to target tissues is restricted. Moreover, because SHBG does not bind NGMN, it appears to be quantitatively the more important NGM metabolite available to target tissues, and probably accounts for a substantial proportion of the progestogenic activity of NGM/EE OCs. However, it is also possible that simultaneous exposure of NGMN and LNG after treatment with NGM/EE may provide clinical benefits not seen with LNG/EE combinations. PMID- 12586320 TI - A study of co-treatment of nonsteroidal anti-inflammatory drugs (NSAIDs) with misoprostol for cervical priming before suction termination of first trimester pregnancy. AB - This double-blind randomized control study was conducted to evaluate whether a nonsteroidal anti-inflammatory drug (NSAID) could act as an effective pain control method during first trimester suction abortion, and whether co-treatment of NSAID with misoprostol will decrease the efficacy of the cervical ripening effect of misoprostol. Subjects were randomized to receive misoprostol alone or misoprostol together with diclofenac sodium. Both groups of subjects suffered from similar incidence of preoperative side effects. Co-treatment of NSAID with misoprostol did not attenuate the cervical ripening efficacy of misoprostol. There was no significant pain reduction in the group treated with NSAID, except that a marginal benefit was found in the subgroup of multiparous women. About two thirds of the subjects in both treatment groups found that this was a satisfactory pain relief method during the procedure. PMID- 12586321 TI - Contraceptive practice before and after termination of pregnancy: a prospective study. AB - This study is part of a larger prospective research program focusing on termination of pregnancy (TOP). One hundred and three women requesting TOP were interviewed before the intervention and 6 months later using open and closed questions and psychological tests. This paper focuses on contraceptive practices before and after abortion. The analysis took into account specific aspects of contraceptive practices and patients' behaviors. The aims were to assess: the level of women's knowledge and practice of contraception at the time of request for a TOP; the behavioral modifications following professional counseling 6 months after TOP; the influence of psychological and sexual factors, and those linked to the women's use of contraception. Most women (n = 101) had already used recommended contraception. During the cycle that had resulted in pregnancy, more than half (n = 58) had used recommended contraception and one third had not used any contraception. Six months later, 86 women used recommended contraception, and 17 did not. The majority of women reported changes in their contraceptive methods (n = 82). Most changes were within recommended methods. The women (n = 10) who continued to practice unprotected intercourse post-TOP were slightly older, satisfied with their sexual relations with their partner, often involved in a long-term and good relationship. During post-TOP period, it is essential to take into account the psychological dynamics involved in the choice of contraceptive methods. Counseling should emphasize not only protection against an unwanted pregnancy but also protection against sexually transmitted diseases, which is often perceived as a less important issue following TOP. PMID- 12586322 TI - Counseling in the clinical setting to prevent unintended pregnancy: an evidence based research agenda. AB - Unintended pregnancies account for about half of all pregnancies in the United States and, in 1995, numbered nearly 3 million pregnancies. They pose appreciable medical, emotional, social and financial costs on women, their families and society. The US is not attaining national goals to decrease unintended pregnancies, and little is known about effective means for reducing unintended pregnancy rates in adults or adolescents.To examine the evidence about the effectiveness, benefits and harms of counseling in a clinical setting to prevent unintended pregnancy in adults and adolescents and to use the evidence to propose a research agenda.We identified English-language articles from comprehensive searches of the MEDLINE, CINAHL, PsychLit and other databases from 1985 through May 2000; the main clinical search terms included pregnancy (mistimed, unintended, unplanned, unwanted), family planning, contraceptive behavior, counseling, sex counseling, and knowledge, attitudes and behavior. We also used published systematic reviews, hand searching of relevant articles, the second Guide to Clinical Preventive Services and extensive peer review to identify important articles not otherwise found and to assure completeness. Of 673 abstracts examined, we retained 354 for full article review; of these, we used 74 for the systematic evidence review and abstracted data from 13 articles for evidence tables. Four studies addressed the effectiveness of counseling in a clinical setting in changing knowledge, skills and attitudes about contraception and pregnancy; all had poor internal validity and generalizability and collectively did not provide definitive guidance about effective counseling strategies. Nine studies (three in teenage populations) addressed the relationship of knowledge on contraceptive use and adherence. Knowledge of correct contraceptive methods may be positively associated with appropriate use, but reservations about the method itself, partner support of the method, and women's beliefs about their own fertility are important determinants of method adherence that may attenuate the knowledge effect. Many factors influence contraceptive use and adherence; among them are age, marital status, ambivalence about becoming pregnant, attitudes of partner, side effects, satisfaction with provider and costs; however, the impact of such factors may not be consistent across populations defined by cultural, age or other factors. The studies themselves differed materially in outcome variables, populations and methodologies and did not yield a body of work that can reliably identify specific influences on contraceptive use and adherence. No literature reports on harms of counseling or on the costs or cost-effectiveness of different approaches to counseling about unintended conceptions in the primary care setting. Virtually no experimental or observational literature reliably answers questions about the effectiveness of counseling in the clinical setting to reduce rates of unintended (unwanted, mistimed) pregnancies in this country. Existing studies suffer from appreciable threats to internal validity and loss to follow-up and are extremely heterogeneous in terms of populations studied and outcomes measured. The quality of the existing research does not provide strong guidance for recommendations about clinical practice but does suggest directions for future investigations. Numerous issues warrant rigorous investigation. PMID- 12586323 TI - Married men's opinions and involvement regarding family planning in rural areas. AB - As men play a prominent role in reproduction, it is therefore extremely useful to assess and encourage them to be involved in contraception, particularly in developing countries, where contraceptive goals have not been reached. This study was carried out in Kayseri, Turkey, in order to determine the attitudes and behavior of married men concerning family planning. A questionnaire was presented to 123 married men. In our study, 91.9% of men approved of family planning, but only 54.4% actually used any contraceptive methods; 66.7% of the men said that the decision should be a joint one, 66.4% wanted to limit their family size. Approximately one fourth of the men had never heard of voluntary sterilization. No one in the study group was aware of the mini-pill, diaphragm or Norplant. In the study group, 26.8% of the men did not want their wives to use intrauterine devices and 31.7% of them did not agree with women using the contraceptive pills. Among those unwilling to use a condom (46.3%), 70.1% stated that it might interrupt intercourse. If a contraceptive pill for males could be used, 25.2% of members of the group would be prepared to use it. Only 17.5% men in the study group had contacted a doctor or a health foundation to obtain information. The main sources of family planning information were TV/radio, followed by friends and newspapers/magazines. In order to encourage men's involvement in family planning, the use of mass media and continual training programs, to try to reach both men and women, could be very useful. PMID- 12586324 TI - Use of prostate-specific antigen (PSA) to measure semen exposure resulting from male condom failures: implications for contraceptive efficacy and the prevention of sexually transmitted disease. AB - Accurate measurement of semen exposure resulting from condom failures can refine public health messages and improve predictions of condom efficacy in preventing pregnancy and HIV transmission. Eight hundred and thirty couples enrolled in a condom efficacy study were asked to collect a baseline sample of ejaculate from the inside of the first study condom they used and to collect a postcoital vaginal sample whenever a study condom broke or slipped off during intercourse. All samples were quantitatively tested for prostate-specific antigen (PSA), a substance found only in human semen, using rocket immunoelectrophoresis, and inspected microscopically for presence of sperm. Sixty-eight baseline ejaculate samples collected from the inside of the first study condom by couples who subsequently experienced a condom failure averaged 13.4 microg PSA per swab and 79% of the samples averaged one or more sperm per high power field (hpf). Seventy nine postcoital vaginal samples obtained after a condom break averaged 5.7 microg PSA per swab and only 38% averaged one or more sperm per hpf. The PSA results indicated a 50% reduction in semen exposure compared to baseline levels (p = 0.0001). Seventeen samples obtained after a condom slip-off averaged 2.5 microg PSA per swab and none of the samples averaged one or more sperm per hpf. The PSA results indicated an 80% reduction in semen exposure compared to baseline levels (p = 0.0001). Our results suggest that even condoms that fail reduce the risk of pregnancy and the transmission of sexually transmitted disease compared to unprotected intercourse. We also used PSA results to adjust a model designed to predict consistent-use pregnancy rates from condom breakage and slippage data. PMID- 12586325 TI - Antifertility effect of an octapeptide, a fragment of FSH binding inhibitor, in the common marmoset (Callithrix jacchus). AB - The follicle-stimulating hormone (FSH) binding inhibitor (FSHBI) has been identified as one of the factors present in follicular fluid exerting autocrine/paracrine effects on FSH actions in the ovary. Our group has isolated FSHBI from human ovarian follicular fluid and deduced its partial amino acid sequence from the N-terminal region. A synthetic peptide corresponding to this sequence also demonstrated FSH binding inhibitory activity in vitro. The objective of the present study was to elucidate the effect of the octapeptide on ovarian cyclicity and pregnancy in the common marmoset. For the study, three groups of postpartum marmosets were treated with the octapeptide during the follicular phase. Administration of the octapeptide from days 6-10 postpartum (pp) predominantly induced luteal insufficiency in two of seven and two of five marmosets at a dose of 100 microg/day (group 1) and 300 microg/day (group 2), respectively. This was confirmed by the presence of small regressing corpora lutea on day 13 pp. Pregnancy was terminated prematurely in one animal from each of these groups. The treatment impaired fertility by 43% and 60% in the marmosets of groups 1 and 2, respectively. In view of the fact that FSH levels peak on day 2 and day 6 of the follicular phase in marmosets, a third group was administered 200 microg/day octapeptide from days 1-8 pp. The treatment induced luteal insufficiency in one out of four marmosets, while premature termination of pregnancy occurred in two other marmosets of this group, demonstrating a 75% effect on pregnancy. Thus, treatment of marmosets with the octapeptide, a fragment of the FSHBI, predominantly induced luteal insufficiency as well as resorption of the fetus leading to impairment of fertility. PMID- 12586326 TI - Human spermatozoa motility analysis in a Ringer's solution containing cupric ions. AB - The ability of cupric ions to immobilize human spermatozoa was assessed in vitro using an optical microscope. A Ringer's solution containing human spermatozoa and three cupric ion concentrations of 8 x 10(-8) mol/L, 8 x 10(-6) mol/L and 8 x 10( 5) mol/L were tested. The higher cupric ion concentrations (8 x 10(-5) mol/L and 8 x 10(-6) mol/L) significantly reduced spermatozoa motility after about 20 min. However, with the lower cupric ion concentration tested (8 x 10(-8) mol/L), also for 20-min experimentation, the reduction in motility was more limited. PMID- 12586327 TI - The tragic matter. PMID- 12586328 TI - Pharmacological evidence for a role of liver X receptors in atheroprotection. PMID- 12586329 TI - T-0901317, a synthetic liver X receptor ligand, inhibits development of atherosclerosis in LDL receptor-deficient mice. AB - Liver X receptors (LXR alpha and LXR beta) are nuclear receptors, which are important regulators of cholesterol and lipid metabolism. LXRs control genes involved in cholesterol efflux in macrophages, bile acid synthesis in liver and intestinal cholesterol absorption. LXRs also regulate genes participating in lipogenesis. To determine whether the activation of LXR promotes or inhibits development of atherosclerosis, T-0901317, a synthetic LXR ligand, was administered to low density lipoprotein receptor (LDLR)(-/-) mice. T-0901317 significantly reduced the atherosclerotic lesions in LDLR(-/-) mice without affecting plasma total cholesterol levels. This anti-atherogenic effect correlated with the plasma concentration of T-0901317, but not with high density lipoprotein cholesterol, which was increased by T-0901317. In addition, we observed that T-0901317 increased expression of ATP binding cassette A1 in the lesions in LDLR(-/-) mice as well as in mouse peritoneal macrophages. T-0901317 also significantly induced cholesterol efflux activity in peritoneal macrophages. These results suggest that LXR ligands may be useful therapeutic agents for the treatment of atherosclerosis. PMID- 12586330 TI - Promoter analysis of a medaka fish intestinal guanylyl cyclase gene. AB - We characterized the promoter activity of a medaka fish intestinal guanylyl cyclase gene, OlGC6, by assay of enzyme activity in response to various promoter luciferase fusion gene constructs introduced into CACO-2 cells and medaka fish embryos. A transient transfection assay of the various fusion gene constructs showed that the nucleotides between -98 and -89 in the 5'-flanking region of the OlGC6 gene are essential for transcription of the OlGC6 gene in CACO-2, and that the OlGC6 gene fragment between -98 and +50 is sufficient to drive gene expression in the medaka fish intestine. An electrophoretic mobility shift assay and ultraviolet (UV) cross-linking experiments demonstrated that a nuclear protein from CACO-2 cells and the adult medaka fish intestinal cells binds specifically to the AGACCTTTGC nucleotides in the regulatory element. PMID- 12586331 TI - Angiostatin and endostatin inhibit endothelial cell migration in response to FGF and VEGF without interfering with specific intracellular signal transduction pathways. AB - The anti-angiogenic agents angiostatin and endostatin have been shown to affect endothelial cell migration in a number of studies. We have examined the effect of these agents on intracellular signalling pathways known to regulate endothelial cell migration and proliferation/survival. Both agents inhibited fibroblast growth factor (FGF)-, and vascular endothelial growth factor (VEGF)-mediated migration of primary human microvascular endothelial cells and affected vascular formation in the embryoid body model. However, using phosphospecific antibodies we could not detect any effect of angiostatin or endostatin on phospholipase C gamma (PLC-gamma), Akt/PKB, p44/42 mitogen-activated protein kinase (MAPK), p38 MAPK and p21-activated kinase (PAK) activity. Furthermore, using a glutathione S transferase (GST)-PAK pull-down assay, we could not detect any effect on Rac activity. We conclude that angiostatin and endostatin inhibit chemotaxis, without affecting intracellular signalling pathways known to regulate endothelial migration and proliferation/survival. PMID- 12586332 TI - Mutational suppression of transferrin receptor shedding can be compensated by distinct metalloproteases acting on alternative sites. AB - The human transferrin receptor (TfR) is proteolytically cleaved at R100 within the juxtamembrane stalk and to a lesser extent at an alternative site. We examined the effect of stalk mutations on human TfR shedding in transfected CHO cells. Point mutations at R100 led to an increase in alternative shedding while the R100 cleavage product was undetectable. Replacing the TfR-stalk by the corresponding sequences from tumor necrosis factor-alpha or interleukin-6 receptor also led to TfR ectodomain shedding. These results show that cleavage at alternative sites can compensate for suppressed cleavage at the major site and inhibitor studies reveal that at least three metalloproteases are involved in the shedding process. PMID- 12586334 TI - Effect of classic preconditioning on the gene expression pattern of rat hearts: a DNA microarray study. AB - To profile gene expression patterns involved in ischemic preconditioning, we monitored global gene expression changes by DNA microarray analysis of 3200 rat specific genes and by real-time quantitative polymerase chain reaction in rat hearts. Forty-nine genes with altered expression were found after ischemia/reperfusion as compared to control non-ischemic hearts and 31 genes were characteristic for classic preconditioning followed by ischemia/reperfusion as compared to ischemia/reperfusion without preconditioning. Genes with altered expression due to ischemia and/or preconditioning included those controlling protein degradation, stress responses, apoptosis, metabolic enzymes, regulatory proteins, and several unknown cellular functions. Metallothionein, natriuretic peptides, coagulation factor VII, cysteine proteinase inhibitor, peroxisome proliferator activator receptor gamma and myosin light chain kinase genes were previously suspected to be related to several cardiovascular diseases, however, most of these genes have not previously been shown to be related to myocardial ischemia/reperfusion. Some genes were observed to change specifically in response to preconditioning: oligoadenylate synthase, chaperonin subunit epsilon, a cGMP phosphodiesterase (PDE9A1), a secretory carrier membrane protein, an amino acid transporter, and protease 28 subunit. None of these genes has previously been shown to be involved in the mechanism of preconditioning. PMID- 12586333 TI - Smad mediates BMP-2-induced upregulation of FGF-evoked PC12 cell differentiation. AB - We previously reported that bone morphogenetic protein (BMP)-2 augments fibroblast growth factor (FGF)-induced neuronal differentiation of PC12 cells by selectively upregulating FGF receptor (FGFR)-1 expression. Here we describe the underlying mechanism. BMP-2 activated Smad proteins in PC12 cells. Overexpression of Smad7 or Smad1, inhibitory and receptor-regulated isoforms, respectively, suppressed or enhanced BMP-2-induced upregulation of FGFR-1 expression. Smad 7 also inhibited the FGF-induced PC12 differentiation. Our findings indicate that activation of a Smad signaling pathway is required for upregulation of FGFR-1 expression by BMP-2 and for the synergistic induction of PC12 differentiation by BMP-2 and FGF. PMID- 12586335 TI - Phosphatase activity of non-heme chloroperoxidase from the bacterium Serratia marcescens. AB - Haloperoxidases are enzymes capable of formation of carbon-halogen bonds in the presence of hydrogen peroxide and halide ions. A mechanism of halogenation catalyzed by heme- and metal-independent bacterial haloperoxidases differs from other representatives of this group of enzymes. Here we report for the first time that bacterial non-heme haloperoxidases possess a phosphatase activity. Chloroperoxidase from Serratia marcescens W 250 purified up to homogeneity is shown to catalyze p-nitrophenylphosphate hydrolysis (K(m) value, 1.8+/-0.1 mM at pH 5.7). The reaction is activated by Mg(2+) and F(-), and is inhibited by WO(4)(2-), tartrate, acetate and phosphate anions. The irreversible inhibition by phenylmethanesulfonyl fluoride, modifier of serine residue in active site, decreases in the presence of phosphate ions. A mechanism of phosphoesters hydrolysis by non-heme haloperoxidases is proposed. PMID- 12586336 TI - Impaired voltage-gated K+ channel expression in brain during experimental cancer cachexia. AB - Cancer-induced cachexia affects most advanced cancer patients. It is characterized by anorexia, profound metabolic dysfunctions, and severe neurological disorders. Here we show that voltage-gated potassium channel (Kv) expression is impaired in the brain of tumor-bearing animals. Expression of both delayed rectifier (Kv1.1, Kv1.2, Kv1.3, Kv1.5, Kv1.6, Kv2.1, Kv3.1, Kv4.2) and A type potassium channels (Kv1.4, Kv3.3, Kv3.4) was greatly down-regulated in brain from animals bearing a Yoshida AH-130 ascites hepatoma. The possible compensatory mechanisms (Kv1.4/Kv4.2), expression of redundant genes (Kv3.1/Kv3.3) and heteromultimeric channel formation (Kv2.1/Kv9.3) were also affected. The high circulating levels of TNFalpha and the reduced expression of the anti-apoptotic protein Bcl-XL found in the brain of tumor-bearing animals indicate that this response could be mediated by an increase in brain cell death due to apoptosis. The results suggest that brain function is impaired during cancer cachexia, and may account for the cancer-induced anorectic response and other neurological alterations. PMID- 12586337 TI - Mitochondrial ATP-sensitive K+ channel opening decreases reactive oxygen species generation. AB - Mitochondrial ATP-sensitive K(+) channel (mitoK(ATP)) opening was shown previously to slightly increase respiration and decrease the membrane potential by stimulating K(+) cycling across the inner membrane. Here we show that mitoK(ATP) opening reduces reactive oxygen species generation in heart, liver and brain mitochondria. Decreased H(2)O(2) release is observed when mitoK(ATP) is active both with respiration stimulated by oxidative phosphorylation and when ATP synthesis is inhibited. In addition, decreased H(2)O(2) release is observed when mitochondrial Delta pH is enhanced, an effect expected to occur when mitoK(ATP) is open. We conclude that mitoK(ATP) is an effective pathway to trigger mild uncoupling, preventing reactive oxygen species release. PMID- 12586338 TI - Peptides derived from the heptad repeat region near the C-terminal of Sendai virus F protein bind the hemagglutinin-neuraminidase ectodomain. AB - Previously, we showed that Sendai virus fusion protein (F) acts as an inhibitor of neuraminidase activity of hemagglutinin-neuraminidase (HN) protein. Here we report that synthetic peptides derived from the heptad repeat region proximal to the transmembrane domain (HR2) of Sendai virus F inhibit fusion and enhance the enzymatic activity of the HN. This occurs on the virus-bound HN and on its soluble globular head. The enhancing effect on virus-bound HN is reversible and depends on the presence of F. The data indicate that, by binding to the HN ectodomain, the HR2 peptides abolish the F inhibition of HN and disrupt the communication between the F and HN essential to promote virus-cell fusion. PMID- 12586339 TI - Prion-like protein Doppel expression is not modified in scrapie-infected cells and in the brains of patients with Creutzfeldt-Jakob disease. AB - Doppel protein has been discovered in prnp knock-out mouse lines, with overproduction of this protein in the brain causing ataxia and neurodegeneration. We investigated whether Doppel expression (i) affected or was affected by the course of prion propagation in neuroblastoma cells, or (ii) modulated Creutzfeldt Jakob disease pathogenesis. No change in Doppel production was detected in N2a cells, before or after infection. Transient murine Doppel gene expression had no effect on N2a viability or PrP(Sc) production. A sensitive immunometric assay revealed low levels of Doppel in human brain, reflecting weak transcription of the corresponding gene. No difference in brain Doppel levels was observed between Creutzfeldt-Jakob disease patients and controls, adding further evidence that Doppel is unlikely to be involved in prion disease pathogenesis. PMID- 12586340 TI - Cyclin B1 transcription is enhanced by the p300 coactivator and regulated during the cell cycle by a CHR-dependent repression mechanism. AB - Cyclin B is a central regulator of transition from the G(2) phase of the cell cycle to mitosis. In mammalian cells two B-type cyclins have been characterised, cyclin B1 and B2. Both are expressed with a maximum in G(2) and their synthesis is mainly regulated on the transcriptional level. We show that a single cell cycle genes homology region, lacking a functional cell cycle-dependent element in tandem with it, contributes most of the cell cycle-dependent transcription from the cyclin B1 promoter. The coactivator p300 binds to the cyclin B1 promoter and synergises with the transcription factor NF-Y in activating transcription of cyclin B1. PMID- 12586341 TI - Requirement of autophosphorylated tyrosine 992 of EGF receptor and its docking protein phospholipase C gamma 1 for membrane ruffle formation. AB - Stimulation of the epidermal growth factor receptor (EGFR) produces membrane ruffles through the small G protein Rac1; however, the signaling pathway from EGFR to Rac1 has not yet been clarified. Here, we show that autophosphorylation of EGFR at tyrosine 992 is required for EGF-induced membrane ruffle formation in CHO cells. Signaling from the autophosphorylated tyrosine 992 appears to be mediated by phospholipase C (PLC) gamma 1. Furthermore, activation of Rac1 by EGF is inhibited by a PLC inhibitor. These results, taken together, suggest that autophosphorylation of EGFR at tyrosine 992 and the subsequent PLC gamma 1 activation transduce the signal to Rac1 to induce membrane ruffle formation. PMID- 12586342 TI - Unexpected catalytic site variation in phosphoprotein phosphatase homologues of cofactor-dependent phosphoglycerate mutase. AB - The cofactor-dependent phosphoglycerate mutase (dPGM) superfamily contains, besides mutases, a variety of phosphatases, both broadly and narrowly substrate specific. Distant dPGM homologues, conspicuously abundant in microbial genomes, represent a challenge for functional annotation based on sequence comparison alone. Here we carry out sequence analysis and molecular modelling of two families of bacterial dPGM homologues, one the SixA phosphoprotein phosphatases, the other containing various proteins of no known molecular function. The models show how SixA proteins have adapted to phosphoprotein substrate and suggest that the second family may also encode phosphoprotein phosphatases. Unexpected variation in catalytic and substrate-binding residues is observed in the models. PMID- 12586343 TI - H9c2 cardiac myoblasts undergo apoptosis in a model of ischemia consisting of serum deprivation and hypoxia: inhibition by PMA. AB - Cardiac myocytes undergo apoptosis under condition of ischemia. Little is known, however, about the molecular pathways that mediate this response. We show that serum deprivation and hypoxia, components of ischemia in vivo, resulted in apoptosis of rat ventricular myoblast cells H9c2. Hypoxia alone did not induce significant apoptosis for at least 48 h, but largely increased the proapoptotic action of serum deprivation. H9c2 cells apoptosis is evidenced by an increase in terminal (TdT)-mediated dUTP nick end-labeling-positive nuclei and by activation of caspases 3, 6, 7 and 9, and loss of mitochondrial functions. In this model of simulated ischemia, represented by serum deprivation plus hypoxia, cardiomyoblasts apoptosis was associated with a p53-independent Bax accumulation and with a down-regulation of Bcl-xL, whereas the levels of cIAP-1, cIAP-2 and X IAP proteins did not change. Phorbol-12-myristate-13-acetate significantly reduced the induction of apoptosis, inhibiting caspase 3 cleavage, Bax accumulation, Bcl-xL down-regulation as well as restoring cell viability. PMID- 12586344 TI - Identification of a mitochondrial glycerol-3-phosphate dehydrogenase from Arabidopsis thaliana: evidence for a mitochondrial glycerol-3-phosphate shuttle in plants. AB - We report molecular characterization of an Arabidopsis gene encoding a mitochondrial FAD-dependent glycerol-3-phosphate dehydrogenase (FAD-GPDH) that oxidizes glycerol-3-phosphate (G-3-P) to dihydroxyacetone phosphate. We demonstrate through in vitro targeting assays that the encoded gene product can be imported into mitochondrial membrane systems. Enzyme activity of the protein was confirmed through heterologous expression in Escherichia coli. The Arabidopsis gene is expressed throughout plant development, but at the highest level during seed germination. We also show that expression of the Arabidopsis FAD-GPDH gene is coupled to oxygen consumption and affected by ABA and stress conditions. Together with an NAD(+)-dependent GPDH, this enzyme could form a G-3 P shuttle, as previously established in other eukaryotic organisms, and links cytosolic G-3-P metabolism to carbon source utilization and energy metabolism in plants. PMID- 12586345 TI - Use of aminoglycoside adenyltransferase translational fusions to determine topology of thylakoid membrane proteins. AB - We have developed a system to examine the topology of thylakoid membrane proteins using the bacterial aadA gene as a reporter. Translational fusions that place the aminoglycoside adenyltransferase domain in the stroma should provide high antibiotic resistance, while those that place it in the thylakoid lumen should give rise to low resistance. Genes encoding chimeric polypeptides consisting of AadA fused to varying lengths of the PsaA polypeptide, whose topology is known, were introduced into the chloroplast genome of Chlamydomonas reinhardtii. As expected, chimeras with an even number of alpha-helices in general resulted in higher resistance. This effect was not due to differences in expression or in catalytic activity. This system should prove useful in analysis of novel proteins predicted to be localized to the thylakoid membrane. PMID- 12586346 TI - Identification of genes possibly related to storage root induction in sweet potato. AB - To identify genes related to initiation of storage root development in sweet potato, a cDNA library was constructed with early stage storage roots (0.3-1 cm in diameter). Single-pass sequences of the 5' ends of 2859 sweet potato cDNA clones were assembled into 483 clusters and 442 singletons. Comparison of sweet potato expressed sequence tags (ESTs) to nodulation/tumorigenesis-related sequence databases (nodule-, tumor-, potato tuber- and development-related sequences) revealed that homologs of 39 sweet potato EST sequences potentially involved in gene regulation, signal transduction and development were present in at least one of the nodulation/tumorigenesis-related sequence databases. Northern blot analyses of these 39 sequences identified 22 differentially expressed genes in early stage storage root and fibrous root. These differentially expressed genes will be potential candidates for research to elucidate the molecular processes related to sweet potato storage root induction. PMID- 12586347 TI - The crystal structure of a plant lectin in complex with the Tn antigen. AB - The structure of the tetrameric Vicia villosa isolectin B4 (VVLB4) in complex with a cancer antigen, the Tn glycopeptide (GalNAc-O-Ser), was determined at 2.7 A resolution. The N-acetylgalactoside moiety of the ligand binds to the primary combining site of VVLB4 in a similar way as observed for other Gal/GalNAc specific plant lectins. The amino acid moiety of the Tn antigen is largely exposed to the solvent and makes few contacts with the protein. The structure of the complex provides a framework to understand the differences in the strength of VVLB4 binding to different sugars and emphasizes the role of a single protein residue, Tyr127, as a structural determinant of Tn-binding specificity. PMID- 12586348 TI - Involvement of NF-Y and Sp1 binding sequences in basal transcription of the human telomerase RNA gene. AB - The proximal promoter of the telomerase RNA gene, hTR, contains four Sp1 sites and one CCAAT box. We have carried out a functional analysis of the role of these sequence elements. Two Sp1 sites downstream of the CCAAT box mediated negative regulation, while the other two Sp1 sites were positive regulators with the strongest effect mediated by the negative regulatory Sp1 site closely flanking the CCAAT box. Basal transcriptional activity is maintained via the CCAAT box even when all four Sp1 sites are mutated, suggesting nuclear factor-Y (NF-Y) is a fundamental regulator of hTR promoter function. Chromatin immunoprecipitation revealed binding of NF-Y, Sp1 and TFIIB to the promoter in vivo. Thus the interaction of NF-Y at the CCAAT box is pivotal to hTR gene transcription and surrounding sequence elements may provide an environment for the regulation of activity through recruitment of additional protein complexes. PMID- 12586349 TI - Highly efficient transport of carboxyfluorescein diacetate succinimidyl ester into COS7 cells using human papillomavirus-like particles. AB - Human papillomavirus virus-like particles (VLPs) have recently been used to deliver genes into mammalian cells in vitro and in vivo. Here, we investigated whether VLPs may serve as an efficient carrier of low molecular weight compounds (e.g. hormones, vitamins, peptides etc.) into cells. COS7 cells were incubated with recombinant HPV-16L1/L2 VLPs labelled with the fluorescence dye carboxyfluorescein diacetate succinimidyl ester. Using flow cytometry, we demonstrate that labelled VLPs can specifically bind to the cell surface followed by their complete internalisation. Our results indicate that VLPs are promising vehicles for highly efficient delivery of low molecular weight compounds into cells. PMID- 12586350 TI - The kinetics of V-J joining throughout 3.5 megabases of the mouse Ig kappa locus fit a constrained diffusion model of nuclear organization. AB - To gain insight into the nuclear organization of the mouse Ig kappa locus and how it may relate to the formation of synapses during recombination, we have studied the kinetics of rearrangement of different V kappa gene families to J kappa gene segments in the pre-B cell line, 103bcl2. Remarkably, V kappa gene families separated by more than 3.5 Mb from J kappa gene segments rearranged with nearly identical kinetics to those as close as 18 kb to J kappa gene segments. These results fit a model of nuclear organization in which the entire V kappa J kappa region resides within a single nuclear subcompartment and is capable of exhibiting multiple reversible contacts through diffusion and Brownian motion. PMID- 12586351 TI - Detection of bound and free IGF-1 and IGF-2 in human plasma via biomolecular interaction analysis mass spectrometry. AB - Insulin like growth factor (IGF)-1 and IGF-2 were assayed from human plasma via biomolecular interaction analysis mass spectrometry, utilizing antibodies as ligands for affinity retrieval. Detection of both targeted and non-targeted IGFs in the mass spectra indicated possible protein complex retrieval by the individual antibodies. A series of control experiments eliminated the possibility of analyte cross-walking between flow cells, significant antibodies cross reactivity, and direct IGF interactions. To disrupt the putative protein complex and release its constituent proteins, plasma samples were treated with detergents. An SDS-treated plasma yielded IGF signals in a different ratio than the one observed in the mass spectra from the non-treated plasma, suggesting disruption of the protein complex, and its retrieval from non-treated plasma. Novel truncated IGF-2 variant, missing its N-terminal Alanine, was detected in all mass spectra. PMID- 12586352 TI - Structure-function analysis of the A20-binding inhibitor of NF-kappa B activation, ABIN-1. AB - Nuclear factor kappa B (NF-kappa B)-dependent gene expression plays an important role in numerous cellular processes including stress responses, inflammation and cell proliferation. Therefore, the activity of this transcription factor needs to be tightly regulated. Among others, the NF-kappa B-dependent zinc finger protein A20 is involved in the negative feedback regulation of NF-kappa B activation in response to tumor necrosis factor (TNF). We previously demonstrated that A20 can interact with A20-binding inhibitors of NF-kappa B activation (ABINs), which have the potential to inhibit TNF-induced activation of NF-kappa B upon overexpression. The ABIN proteins were therefore proposed to mediate the NF-kappa B inhibiting function of A20. Here we demonstrate the presence of a short homologous region in ABINs and I kappa B kinase gamma, the regulatory subunit of the I kappa B kinase complex. Site-specific mutagenesis of this region abolished the NF-kappa B inhibiting function of ABIN-1, without affecting the interaction with A20. Furthermore, coexpression of these ABIN-1 mutants interfered in a dominant negative manner with the NF-kappa B inhibiting function of ABIN-1, whereas the A20-mediated inhibition was unaffected. These results suggest that A20 and ABIN-1 probably act independently of their mutual interaction. PMID- 12586353 TI - NHERF-1 uniquely transduces the cAMP signals that inhibit sodium-hydrogen exchange in mouse renal apical membranes. AB - Sodium-hydrogen exchanger regulatory factor isoform-1 (NHERF-1) and NHERF-2 are two structurally related PDZ-domain-containing protein adapters that effectively transduce cyclic AMP (cAMP) signals that inhibit NHE3, the sodium-hydrogen exchanger isoform present at the apical surface of kidney and gut epithelia. The mouse renal proximal tubule expresses both NHERF isoforms, suggesting their redundant functions as regulators of renal electrolyte metabolism. To define the role of NHERF-1 in the physiological control of NHE3, we analyzed NHE3 activity in isolated brush border membrane (BBM) preparations from renal proximal tubules of wild-type (WT) and NHERF-1 (-/-) mice. Basal Na(+)-H(+) exchange was indistinguishable in BBMs from WT and NHERF-1 (-/-) mice (0.96+/-0.08 and 0.95+/ 0.10 nmol/mg protein/10 s, respectively). Activation of membrane bound cAMP dependent protein kinase (PKA) by cAMP inhibited NHE3 activity in WT BBMs (0.55+/ 0.07 nmol/mg protein/10 s or 40+/-9%, P<0.01) but had no discernible effect on Na(+)-H(+) exchange in the NHERF-1 (-/-) BBM (0.97+/-0.07 nmol/mg protein/10 s; P=not significant). This was associated with a significant decrease in cAMP stimulated phosphorylation of NHE3 immunoprecipitated from solubilized NHERF-1 ( /-) BBMs. As the protein levels for NHE3, NHERF-2, PKA and ezrin were not changed in the NHERF-1 (-/-) BBMs, the data suggest a unique role for NHERF-1 in cAMP mediated inhibition of NHE3 activity in the renal proximal tubule of the mouse. PMID- 12586355 TI - A modified sensor chip for surface plasmon resonance enables a rapid determination of sequence specificity of DNA-binding proteins. AB - A novel method is described which rapidly determines specificity of DNA-binding proteins using a surface plasmon resonance (SPR) sensor chip. An oligohistidine tagged DNA-binding domain of a transcription factor, NtERF2, was immobilised via nitrilotriacetic acid ligands to a sensor chip with an attenuated degree of carboxymethylation. DNA molecules were selected from a pool of randomised oligomers through binding to the immobilised protein and amplified by PCR. After several cycles of selection, during which binding was monitored by SPR, DNA sequences containing a consensus sequence were determined. The time necessary for one cycle is approximately 50 min, which is shorter than existing methods. PMID- 12586354 TI - The hypolipidemic drug metabolites nafenopin-CoA and ciprofibroyl-CoA are competitive P2Y1 receptor antagonists. AB - Coenzyme A (CoA-SH), endogenous and drug-derived CoA-derivatives were tested as putative antagonists of P2Y receptors expressed in Xenopus laevis oocytes, a method used to determine calcium-activated chloride current, an indicator of the activation of these receptors. CoA-SH antagonized reversibly and in a concentration-dependent manner the ATP-gated currents evoked by the human P2Y(1) but not the P2Y(2) receptor. Palmitoyl-CoA was four-fold more potent than CoA-SH as an antagonist while palmitoyl-carnitine was inactive, highlighting the role of the CoA-SH moiety in the antagonism. The CoA derivatives of nafenopin and ciprofibrate, two clinically relevant hypolipidemic drugs, increased 13 and three fold the potency of CoA-SH, respectively. The K(B)s of nafenopin-CoA and ciprofibroyl-CoA were 58 and 148 nM, respectively; the slopes of the Schild plots were unitary. Neither 100 microM nafenopin nor ciprofibrate alone altered the P2Y(1) receptor activity. Neither CoA-SH nor ciprofibroyl-CoA antagonized the rat P2X(2) or the P2X(4) nucleotide receptors nor interacted with the 5-HT(2A/C) receptors. The bulky drug CoA-SH derivatives identify a hydrophobic pocket, which may serve as a potential target for novel selective P2Y(1) antagonists. PMID- 12586356 TI - BML-190 and AM251 act as inverse agonists at the human cannabinoid CB2 receptor: signalling via cAMP and inositol phosphates. AB - The aminoalkylindole BML-190 and diarylpyrazole AM251 ligands have previously been shown to bind to cannabinoid CB(2) and CB(1) receptors, respectively. In HEK 293 cells stably expressing the human CB(2) receptor, BML-190 and AM251 potentiated the forskolin-stimulated accumulation of cAMP. Moreover, the CB(2) receptor can interact productively with 16z44, a promiscuous G alpha(16/z) chimera. BML-190 and AM251 reduce the basal levels of inositol phosphate production in cells expressing the CB(2) receptor and 16z44. These results demonstrate that BML-190 and AM251 act as inverse agonists at the human CB(2) receptor acting via G alpha(i/o) and G alpha(q) family-coupled pathways. PMID- 12586357 TI - Essential role of protein kinase C zeta in the impairment of insulin-induced glucose transport in IRS-2-deficient brown adipocytes. AB - Insulin receptor substrate-2-deficient (IRS-2(-/-)) mice develop type 2 diabetes. We have investigated the molecular mechanisms by which IRS-2(-/-) immortalized brown adipocytes showed an impaired response to insulin in inducing GLUT4 translocation and glucose uptake. IRS-2-associated phosphatidylinositol 3-kinase (PI 3-kinase) activity was blunted in IRS-2(-/-) cells, total PI 3-kinase activity being reduced by 30%. Downstream, activation of protein kinase C (PKC) zeta was abolished in IRS-2(-/-) cells. Reconstitution with retroviral IRS-2 restores IRS-2/PI 3-kinase/PKC zeta signalling, as well as glucose uptake. Wild type cells expressing a kinase-inactive mutant of PKC zeta lack GLUT4 translocation and glucose uptake. Our results support the essential role played by PKC zeta in the insulin resistance and impaired glucose uptake observed in IRS 2-deficient brown adipocytes. PMID- 12586358 TI - A morphologic study of filamentous phage infection of Escherichia coli using biotinylated phages. AB - Using biotinylated phage (BIO-phages), we observed the infection of filamentous phages into Escherichia coli JM109 morphologically. BIO-phages and BIO-phage derived proteins, mainly pVIII, were detected in E. coli by using the avidin biotin-peroxidase complex method with electron microscopy. Infected cells revealed positive staining on the outer and inner membranes and in the periplasmic space. Some cells showed specific or predominant staining of the outer membrane, whereas others showed predominant staining of the inner membrane or equivalent staining of the outer and inner membranes. The periplasmic spaces in some infected cells were expanded and filled with reaction products. Some cells showed wavy lines of positive staining in the periplasmic space. BIO-phages were detected as thick filaments or clusters covered with reaction products. The ends of the infecting phages were located on the surface of cells, in the periplasmic space, or on the inner membrane. These findings suggest that phage major coat proteins are integrated into the outer membrane and that phages cause periplasmic expansion during infection. PMID- 12586359 TI - Ghrelin and growth hormone secretagogue receptor are expressed in the rat adrenal cortex: Evidence that ghrelin stimulates the growth, but not the secretory activity of adrenal cells. AB - Ghrelin is an endogenous ligand of the growth hormone secretagogue receptor (GHS R), which has been originally isolated from rat stomach. Evidence has been previously provided that adrenal gland possesses abundant ghrelin-displaceable GHS-Rs, but nothing is known about the possible role of ghrelin in the regulation of adrenocortical function. Reverse transcription-polymerase chain reaction demonstrated the expression of ghrelin and GHS-R in the rat adrenal cortex, and high adrenal concentrations of immunoreactive ghrelin were detected by radioimmune assay (RIA). Autoradiography localized abundant [(125)I]ghrelin binding sites in the adrenal zona glomerulosa (ZG) and outer zona fasciculata (ZF). Ghrelin (from 10(-10) to 10(-8) M) did not affect either basal steroid hormone (pregnenolone, progesterone, 11-deoxycorticosterone, corticosterone, 18 hydroxycorticosterone and aldosterone) secretion from dispersed ZG and zona fasciculata/reticularis (ZF/R) cells (as evaluated by quantitative high pressure liquid chromatography), or basal and agonist-stimulated aldosterone and corticosterone production from cultured ZG and ZF/R cells, respectively (as measured by RIA). Ghrelin (10(-8) and 10(-6) M) raised basal, but not agonist stimulated, proliferation rate of cultured ZG cells (percent of cells able to incorporate 5-bromo-2'-deoxyuridine), without affecting apoptotic deletion rate (percent of cells able to incorporate biotinylated nucleosides into apoptotic DNA fragments). The tyrosine kinase (TK) inhibitor tyrphostin-23 and the p42/p44 mitogen-activated protein kinase (MAPK) inhibitor PD-98059 abolished the proliferogenic effect of 10(-8) M ghrelin, while the protein kinase A and C inhibitors H-89 and calphostin-C were ineffective. Ghrelin (10(-8) M) stimulated TK and MAPK activity of dispersed ZG cells, and the effect was abolished by preincubation with tyrphostin-23 and PD-98059, respectively. Tyrphostin-23 annulled ghrelin-induced activation of MAPK activity. Taken together, the present findings indicate that (i) ghrelin and GHS-R are both expressed in the rat adrenal cortex, ghrelin binding sites being very abundant in the ZG; (ii) ghrelin does not affect the secretory activity of rat adrenocortical cells, but significantly enhances the proliferation rate of cultured ZG cells, without affecting apoptotic deletion rate; and (iii) the ZG proliferogenic action of ghrelin involves the TK-dependent activation of the p42/p44 MAPK cascade. PMID- 12586360 TI - Regulation of telomerase activity and anti-apoptotic function by protein-protein interaction and phosphorylation. AB - The enzyme telomerase is necessary for the synthesis and maintenance of telomere length. The catalytic subunit, telomerase reverse transcriptase (TERT), is regulated by interaction with the 90 kDa heat shock protein (HSP90) and by Akt dependent phosphorylation. Here, we demonstrate that HSP90 and Akt physically interact with TERT. Treatment of cells with novobiocin, which blocks C-terminal interaction of HSP90, disrupted HSP90 binding to Akt, induced Akt dephosphorylation and significantly reduced telomerase activity. The reduction of TERT activity by novobiocin was associated with an increase in apoptosis. Likewise, the induction of Akt dephosphorylation by protein phosphatase 2A (PP2A) reduced telomerase activity. HSP90 is known to prevent PP2A-mediated dephosphorylation of Akt. To investigate whether the effect of novobiocin is due to the reduction of Akt or TERT phosphorylation, we overexpressed a phospho mimetic, active Akt (T308D/S473D). Akt (T308D/S473D) prevented novobiocin-induced reduction of telomerase activity and the stimulation of apoptosis. Moreover, overexpression of a dominant negative PP2A construct (PP2A(L199P)) as well as incubation with the PP2A inhibitor okadaic acid blocked the inhibition of telomerase activity by novobiocin. These data suggest that the association between HSP90, Akt and TERT in concert with the phosphorylation of TERT is necessary for maintaining telomerase activity and inhibition of apoptosis. PMID- 12586361 TI - Protein folding: could hydrophobic collapse be coupled with hydrogen-bond formation? AB - A judicious examination of an exhaustive PDB sample of soluble globular proteins of moderate size (N<102) reveals a commensurable relationship between hydrophobic surface burial and number of backbone hydrogen bonds. An analysis of 50,000 conformations along the longest all-atom MD trajectory allows us to infer that not only the hydrophobic collapse is concurrent with the formation of backbone amide-carbonyl hydrogen bonds, they are also dynamically coupled processes. In statistical terms, hydrophobic clustering of the side chains is inevitably conducive to backbone burial and the latter process becomes thermodynamically too costly and kinetically unfeasible without amide-carbonyl hydrogen-bond formation. Furthermore, the desolvation of most hydrogen bonds is exhaustive along the pathway, implying that such bonds guide the collapse process. PMID- 12586362 TI - Calcium regulates the PI3K-Akt pathway in stretched osteoblasts. AB - Mechanical loading plays a vital role in maintaining bone architecture. The process by which osteoblasts convert mechanical signals into biochemical responses leading to bone remodeling is not fully understood. The earliest cellular response detected in mechanically stimulated osteoblasts is an increase in intracellular calcium concentration ([Ca(2+)](i)). In this study, we used the clonal mouse osteoblast cell line MC3T3-E1 to show that uniaxial cyclic stretch induces: (1) an immediate increase in [Ca(2+)](i), and (2) the phosphorylation of critical osteoblast proteins that are implicated in cell proliferation, gene regulation, and cell survival. Our data suggest that cyclic stretch activates the phosphoinositide 3-kinase (PI3K) pathway including: PI3K, Akt, FKHR, and AFX. Moreover, cyclic stretch also causes the phosphorylation of stress-activated protein kinase/c-Jun N-terminal kinase. Attenuation in the level of phosphorylation of these proteins was observed by stretching cells in Ca(2+)-free medium, using intra- (BAPTA-AM) and extracellular (BAPTA) calcium chelators, or gadolinium, suggesting that influx of extracellular calcium plays a significant role in the early response of osteoblasts to mechanical stimuli. PMID- 12586363 TI - Na+,K+-ATPase trafficking in skeletal muscle: insulin stimulates translocation of both alpha 1- and alpha 2-subunit isoforms. AB - We determined insulin-stimulated Na(+),K(+)-ATPase isoform-specific translocation to the skeletal muscle plasma membrane. When rat muscle plasma membrane fractions were isolated by discontinuous sucrose gradients, insulin-stimulated translocation of alpha(2)- but not alpha(1)-subunits was detected. However, using cell surface biotinylation techniques, an insulin-induced membrane translocation of both alpha(1) and alpha(2)-subunits in rat epitrochlearis muscle and cultured human skeletal muscle cells was noted. Na(+),K(+)-ATPase alpha-subunit translocation was abolished by the phosphatidylinositol (PI) 3-kinase inhibitor wortmannin, as well as by the protein kinase C inhibitor GF109203X. Thus, insulin mediates Na(+),K(+)-ATPase alpha(1)- and alpha(2)-subunit translocation to the skeletal muscle plasma membrane via a PI 3-kinase-dependent mechanism. PMID- 12586364 TI - Novel insights into cadherin processing by subtilisin-like convertases. AB - Proprotein convertases (PCs) are known to activate many important molecules and their overexpression plays a significant role in tumor progression. Only little is known about the involvement of PCs in the processing of cadherin adhesion molecules, which are potent tumor suppressors. Here we show in a baculovirus overexpression system that the desmosomal cadherins Dsg1 and Dsg3 are substrates for the PC furin. Accordingly, inhibition of PCs in differentiating mouse keratinocytes by alpha 1-anti-trypsin Portland (alpha 1-PDX) negatively interfered with pro-epithelial (proE)-cadherin processing, but unexpectedly also resulted in a dramatic reduction of E-cadherin, Dsg1 and Dsg3 protein and Dsg1 mRNA. Because loss of intercellular adhesion is a rate-limiting step in the transition from benign to malignant tumors, these results have significant implications for the use of PC inhibitors as possible therapeutic tools. PMID- 12586365 TI - Expression of syntaxin 1C, an alternative splice variant of HPC-1/syntaxin 1A, is enhanced by phorbol-ester stimulation in astroglioma: participation of the PKC signaling pathway. AB - Syntaxin 1C is an alternative splice variant of HPC-1/syntaxin 1A; the latter participates in neurotransmitter release and is assigned to the gene domain responsible for Williams' syndrome (WS). It is expressed in the soluble fraction extracted from human astroglioma cell lines T98G and U87MG. Quantitative immunoblot and indirect immunofluorescence analyses revealed that the expression of syntaxin 1C was upregulated by phorbol 12-myristate 13-acetate (PMA), but not by forskolin. A protein kinase C (PKC) inhibitor suppressed this enhancement. These results suggest that syntaxin 1C expression is regulated via the PKC signal pathway. This is the first report of a signal transduction system that directly affects the expression of syntaxin protein. PMID- 12586366 TI - Synthetic and structural studies on Pyrularia pubera thionin: a single-residue mutation enhances activity against Gram-negative bacteria. AB - The thionin from Pyrularia pubera (Pp-TH), a 47-residue peptide with four internal disulfide bonds, was efficiently produced by chemical synthesis. Its antimicrobial activity in vitro against several representative pathogens (EC(50)=0.3-3.0 microM) was identical to that of natural Pp-TH. This peptide has a unique Asp(32) instead of the consensus Arg found in other thionins of the same family. In order to evaluate the effect of this mutation, the Arg(32) analogue (Pp-TH(D32R)) was also synthesized and showed a significant increase in antibiotic activity against several Gram-negative bacteria, whereas it retained the same activity against other pathogens. The overall structure of Pp-TH(D32R) was maintained, though a slight decrease in the helical content of the peptide was observed. PMID- 12586367 TI - Differences in the ubiquitination of p53 by Mdm2 and the HPV protein E6. AB - The human papillomavirus (HPV) protein E6 can promote the ubiquitination of the p53 tumour suppressor in vitro, providing an explanation for the ability of E6 to induce p53 degradation in vivo and contribute to the potential tumorigenic effect of the virus. Instead, in non-infected cells, p53 levels are primarily destabilised by the ubiquitin E3 ligase activity of the Mdm2 protein. Here we have compared the effects of E6 and Mdm2 on p53 ubiquitination in vivo. We show that whereas in the presence of Mdm2 proteasome inhibitors induce the accumulation of ubiquitinated forms of p53, this does not occur in the presence of E6. Accordingly, we confirm that the effect of E6 and p53 is independent of the six C-terminal lysine residues in p53, which have previously been described to play an important role for effective ubiquitination and degradation of 53 mediated by Mdm2. We also show that other yet unidentified residues in p53 are also susceptible to ubiquitination. These results indicate that E6 does not induce ubiquitination of p53 in the same way as Mdm2 in order to promote its degradation, suggesting important differences between the Mdm2 and E6 effects on p53 degradation. PMID- 12586368 TI - Involvement of cholesterol in the inhibitory effect of dimethyl-beta-cyclodextrin on P-glycoprotein and MRP2 function in Caco-2 cells. AB - We compared the inhibitory effect of various cyclodextrins (CyDs) on P glycoprotein (P-gp) and multidrug resistance-associated protein 2 (MRP2) function and examined the contribution of cholesterol to the inhibitory effect of 2,6-di-O methyl-beta-cyclodextrin (DM-beta-CyD) on the efflux activity of the function in Caco-2 cell monolayers. Of various CyDs, DM-beta-CyD significantly impaired the efflux activity of P-gp and MRP2. DM-beta-CyD released P-gp and MRP2 from the monolayers in the apical side's transport buffer and decreased the extent of cholesterol as well as P-gp and MRP2 in caveolae of Caco-2 cell monolayers, but not caveolin and flotillin-1. On the other hand, DM-beta-CyD did not change MDR1 and MRP2 mRNA levels. Therefore, these results suggest that the inhibitory effect of DM-beta-CyD on P-gp and MRP2 function, at least in part, could be attributed to the release of these transporters from the apical membranes into the medium as secondary effects through cholesterol-depletion in caveolae after treatment of Caco-2 cell monolayers with DM-beta-CyD. PMID- 12586369 TI - A forkhead transcription factor FKHR up-regulates lipoprotein lipase expression in skeletal muscle. AB - Lipoprotein lipase (LPL) plays a role in lipid usage in skeletal muscle by hydrolyzing plasma triglycerides into fatty acids, which are further utilized for beta-oxidation. Lipid usage is stimulated during fasting, diabetes mellitus and exercise, concomitant with enhanced LPL expression in skeletal muscle. Here we show that the forkhead type transcription factor FKHR is strongly induced in skeletal muscle in fasting mice, in mice with streptozotocin-induced diabetes and in mice after treadmill running. Ectopic expression of FKHR enhanced LPL gene expression in C2C12 muscle cells in culture. These results implicate FKHR as an important modulator of lipid metabolism in skeletal muscle. PMID- 12586370 TI - Dynamic structure of pharaonis phoborhodopsin (sensory rhodopsin II) and complex with a cognate truncated transducer as revealed by site-directed 13C solid-state NMR. AB - We have recorded (13)C nuclear magnetic resonance (NMR) spectra of [3-(13)C]Ala, [1-(13)C]Val-labeled pharaonis phoborhodopsin (ppR or sensory rhodopsin II) incorporated into egg PC (phosphatidylcholine) bilayer, by means of site-directed high-resolution solid-state NMR techniques. Seven (13)C NMR signals from transmembrane alpha-helices were resolved for [3-(13)C]Ala-ppR at almost the same positions as those of bacteriorhodopsin (bR), except for the suppressed peaks in the loop regions in spite of the presence of at least three Ala residues. In contrast, (13)C NMR signals from the loops were visible from [1-(13)C]Val-ppR but their peak positions of the transmembrane alpha-helices are not always the same between ppR and bR. The motional frequency of the loop regions in ppR was estimated as 10(5) Hz in view of the suppressed peaks from [3-(13)C]Ala-ppR due to interference with proton decoupling frequency. We found that conformation and dynamics of ppR were appreciably altered by complex formation with a cognate truncated transducer pHtr II (1-159). In particular, the C-terminal alpha-helix protruding from the membrane surface is involved in the complex formation and subsequent fluctuation frequency is reduced by one order of magnitude. PMID- 12586371 TI - Activation of NF-kappa B transcription factor in human neutrophils by sulphatides and L-selectin cross-linking. AB - Sulphated galactocerebroside (sulphatide) has been established as a ligand for L selectin and shown to trigger intracellular signals in human neutrophils. We have found that sulphatide activated transcription factor NF-kappa B in human neutrophils in a concentration-dependent manner whereas non-sulphated galactocerebroside did not demonstrate such an effect. The activation was inhibitable by pretreatment with primary monoclonal anti-L-selectin antibody (clone LAM1-116). Binding of the primary antibody to L-selectin was insufficient to induce NF-kappa B activation but cross-linking of L-selectin with a secondary antibody was effective. alpha-Chymotrypsin, the agent known to shed L-selectin, activated NF-kappa B by itself. The response to sulphatides was inhibited by jasplakinolide, an actin-polymerising agent known to downregulate surface expression of L-selectin, Fc gamma RIIIb, CD43 and CD44. Recently we have reported that sulphatide stimulated the attachment of human neutrophils to collagen via Mac1 (CD11b/CD18) integrin [Sud'ina et al., Biochem. J. 359 (2001) 621-629]. We now show signalling from sulphatide to NF-kappa B activation and discuss its involvement in neutrophil adhesion. PMID- 12586374 TI - Assembly and overexpression of membrane proteins in Escherichia coli. AB - The bacterium Escherichia coli is one of the most popular model systems to study the assembly of membrane proteins of the so-called helix-bundle class. Here, based on this system, we review and discuss what is currently known about the assembly of these membrane proteins. In addition, we will briefly review and discuss how E. coli has been used as a vehicle for the overexpression of membrane proteins. PMID- 12586375 TI - Specific lipid requirements of membrane proteins--a putative bottleneck in heterologous expression. AB - Membrane proteins are mostly protein-lipid complexes. For more than 30 examples of membrane proteins from prokaryotes, yeast, plant and mammals, the importance of phospholipids and sterols for optimal activity is documented. All crystallized membrane protein complexes show defined lipid-protein contacts. In addition, lipid requirements may also be transitory and necessary only for correct folding and intercellular transport. With respect to specific lipid requirements of membrane proteins, the phospholipid and glycolipid as well as the sterol content of the host cell chosen for heterologous expression should be carefully considered. The lipid composition of bacteria, archaea, yeasts, insects,Xenopus oocytes, and typical plant and mammalian cells are given in this review. A few examples of heterologous expression of membrane proteins, where problems of specific lipid requirements have been noticed or should be thought of, have been chosen. PMID- 12586376 TI - Practical aspects of overexpressing bacterial secondary membrane transporters for structural studies. AB - Membrane transporter proteins play critical physiological roles in the cell and constitute 5-10% of prokaryotic and eukaryotic genomes. High-resolution structural information is essential for understanding the functional mechanism of these proteins. A prerequisite for structural study is to overexpress such proteins in large quantities. In the last few years, over 20 bacterial membrane transporters were overexpressed at a level of 1 mg/l of culture or higher, most often in Escherichia coli. In this review, we analyzed those factors that affect the quantity and quality of the protein produced, and summarized recent progress in overexpression of membrane transporters from bacterial inner membrane. Rapid progress in genome sequencing provides opportunities for expressing several homologues and orthologues of the target protein simultaneously, while the availability of various expression vectors allows flexible experimental design. Careful optimization of cell culture conditions can drastically improve the expression level and homogeneity of the target protein. New sample preparation techniques for mass spectrometry of membrane proteins have enabled one to identity the rigid protein core, which can be subsequently overexpressed. Size exclusion chromatography on HPLC has proven to be an efficient method in screening detergent, pH an other conditions required for maintaining the stability and monodispersity of the protein. Such high-quality preparations of membrane transporter proteins will probably lead to successful crystallization and structure determination of these proteins in the next few years. PMID- 12586377 TI - The expression of outer membrane proteins for crystallization. AB - The production of sufficient amounts of chemically and conformationally homogenous protein is a major requirement for successful crystallization and structure determination. With membrane proteins, this constitutes a particular problem because the membrane volume is limited and the organisms are usually very sensitive to changes in membrane properties brought about by massive protein insertion. Moreover, the extraction of membrane proteins from the membrane with detergents is generally a harsh treatment, which gives rise to conformational aberrations. A number of successful procedures for functional expression followed by purification are reviewed here together with nonfunctional expression into inclusion bodies and subsequent (re)folding to produce functional proteins. Most of the data are for prokaryotic outer membrane proteins, but the outer membrane proteins of eukaryotic organelles are also considered as they do show similar features. PMID- 12586378 TI - Monoclonal antibodies for the structural analysis of the Na+/H+ antiporter NhaA from Escherichia coli. AB - Since their advent some 25 years ago, monoclonal antibodies have developed into powerful tools for structural and functional analysis of their cognate antigens. Together with the respective antigen binding fragments, antibodies offer exclusive capacities in detection, characterization, purification and functional assays for every given ligand. Antibody-fragment mediated crystallization represents a major advance in determining the three-dimensional structure of membrane-bound protein complexes. In this review, we focus on the methods used to generate monoclonal antibodies against the NhaA antiporter from Escherichia coli as a paradigm of secondary transporters. We describe examples on how antibodies are helpful in understanding structure and function relationships for this important class of integral membrane proteins. The generated conformation specific antibody fragments are highly valuable reagents for co-crystallization attempts and structure determination of the antiporter. PMID- 12586380 TI - In vitro folding of alpha-helical membrane proteins. AB - For large-scale production, as required in structural biology, membrane proteins can be expressed in an insoluble form as inclusion bodies and be refolded in vitro. This requires refolding conditions where the native form is thermodynamically stable and where nonproductive pathways leading to aggregation are avoided. Examples of successful refolding are reviewed and general guidelines to establish refolding protocols of membrane proteins are presented. PMID- 12586379 TI - The trials and tribulations of membrane protein folding in vitro. AB - Membrane proteins are hard to handle and consequently the purification of functional protein in milligram quantities is a major problem. One reason for this is that once integral membrane proteins are outside their native membrane, they are prone to aggregation, are unstable and are frequently only partially functional. Knowledge of membrane protein folding mechanisms in vitro can help to understand the causes of these problems and work toward strategies to disaggregate and fold proteins correctly. Kinetic and stability studies are emerging on membrane protein folding, mainly on bacterial proteins. Mutagenesis methods have also been used to probe specific structural features or bonds in proteins. In addition, manipulation of lipid properties can be used to improve the efficiency of folding as well as the stability and function of the protein. PMID- 12586381 TI - Overexpression, purification, and functional characterization of ATP-binding cassette transporters in the yeast, Pichia pastoris. AB - The ATP-binding cassette (ABC) transporter superfamily is a large gene family that has been highly conserved throughout evolution. The physiological importance of these membrane transporters is highlighted by the large variety of substrates they transport, and by the observation that mutations in many of them cause heritable diseases in human. Likewise, overexpression of certain ABC transporters, such as P-glycoprotein and members of the multidrug resistance associated protein (MRP) family, is associated with multidrug resistance in various cells and organisms. Understanding the structure and molecular mechanisms of transport of the ABC transporters in normal tissues and their possibly altered function in human diseases requires large amounts of purified and active proteins. For this, efficient expression systems are needed. The methylotrophic yeast Pichia pastoris has proven to be an efficient and inexpensive experimental model for high-level expression of many proteins, including ABC transporters. In the present review, we will summarize recent advances on the use of this system for the expression, purification, and functional characterization of P glycoprotein and two members of the MRP subfamily. PMID- 12586382 TI - G protein-coupled receptor overexpression with the baculovirus-insect cell system: a tool for structural and functional studies. AB - G protein-coupled receptors, whose topology shows seven transmembrane domains, form the largest known family of receptors involved in higher organism signal transduction. These receptors are generally of low natural abundance and overexpression is usually a prerequisite to their structural or functional characterisation. The baculovirus-insect cell system constitutes a versatile tool for the maximal production of receptors. This heterologous expression system also provides interesting alternatives for receptor functional studies in a well controlled cellular context. PMID- 12586383 TI - Semliki Forest virus vectors for rapid and high-level expression of integral membrane proteins. AB - Semliki Forest virus (SFV) vectors have been applied for the expression of recombinant integral membrane proteins in a wide range of mammalian host cells. More than 50 G protein-coupled receptors (GPCRs), several ion channels and other types of transmembrane or membrane-associated proteins have been expressed at high levels. The establishment of large-scale SFV technology has facilitated the production of large quantities of recombinant receptors, which have then been subjected to drug screening programs and structure-function studies on purified receptors. The recent Membrane Protein Network (MePNet) structural genomics initiative, where 100 GPCRs are overexpressed from SFV vectors, will further provide new methods and technologies for expression, solubilization, purification and crystallization of GPCRs. PMID- 12586384 TI - Lactococcus lactis as host for overproduction of functional membrane proteins. AB - Lactococcus lactis has many properties that are ideal for enhanced expression of membrane proteins. The organism is easy and inexpensive to culture, has a single membrane and relatively mild proteolytic activity. Methods for genetic manipulation are fully established and a tightly controlled promoter system is available, with which the level of expression can be varied with the inducer concentration. Here we describe our experiences with lactococcal expression of the mechanosensitive channel, the human KDEL receptor and transporters belonging to the ABC transporter family, the major facilitator superfamily, the mitochondrial carrier family and the peptide transporter family. Previously published expression studies only deal with the overexpression of prokaryotic membrane proteins, but in this paper, experimental data are presented for the overproduction of mitochondrial and hydrogenosomal carriers and the human KDEL receptor. These eukaryotic membrane proteins were expressed in a functional form and at levels amenable to structural work. PMID- 12586385 TI - Downstream coding region determinants of bacterio-opsin, muscarinic acetylcholine receptor and adrenergic receptor expression in Halobacterium salinarum. AB - The aim of this work is to develop a prokaryotic system capable of expressing membrane-bound receptors in quantities suitable for biochemical and biophysical studies. Our strategy exploits the endogenous high-level expression of the membrane protein bacteriorhodopsin (BR) in the Archaeon Halobacterium salinarum. We attempted to express the human muscarinic acetylcholine (M(1)) and adrenergic (a2b) receptors by fusing the coding region of the m1 and a2b genes to nucleotide sequences known to direct bacterio-opsin (bop) gene transcription. The fusions included downstream modifications to produce non-native carboxyl-terminal amino acids useful for protein identification and purification. bop mRNA and BR accumulation were found to be tightly coupled and the carboxyl-terminal coding region modifications perturbed both. m1 and a2b mRNA levels were low, and accumulation was sensitive to both the extent of the bop gene fusion and the specific carboxyl-terminal coding sequence modifications included. Functional a2b adrenergic receptor expression was observed to be dependent on the downstream coding region. This work demonstrates that a critical determinant of expression resides in the downstream coding region of the wild-type bop gene and manipulation of the downstream coding region of heterologous genes may affect their potential for expression in H. salinarum. PMID- 12586386 TI - Calnexin co-expression and the use of weaker promoters increase the expression of correctly assembled Shaker potassium channel in insect cells. AB - Voltage-gated potassium channels control the membrane potential of excitable cells. To understand their function, knowledge of their structure is essential. However, these channels are scarce in natural sources, and overexpression is necessary to generate material for structural studies. We have compared functional expression of the Drosophila Shaker H4 potassium channel in stable insect cell lines and in baculovirus-infected insect cells, using three different baculovirus promoters. Stable insect cell lines expressed correctly assembled channel, which was glycosylated and found predominantly at, or close to, the cell surface. In comparison, the majority of baculovirus-overexpressed Shaker was intracellular and incorrectly assembled. The proportion of functional Shaker increased, however, if the weaker basic protein promoter was used rather than the stronger p10 or polyhedrin promoters. In addition, co-expression of the molecular chaperone, calnexin, increased the quantity of correctly assembled channel protein, suggesting that calnexin can be used to increase the efficiency of channel expression in insect cells. PMID- 12586387 TI - Expression and purification of truncated, non-glycosylated turkey beta-adrenergic receptors for crystallization. AB - In order to purify milligram quantities of turkey beta-adrenergic receptor (betaAR) for structural analysis, we have expressed mutant betaARs using the baculovirus system. The initial betaAR construct was truncated at both N- and C termini thus removing an N-glycosylation site. Cys 116 was mutated to leucine and a histidine tag was added at the C-terminus resulting in the betaAR construct 20 424/His6. Expression of this construct in Sf9 cells produced 0.5 mg of unpurified receptor per liter of culture which necessitated the use of a fermenter for large scale production. The yield was improved more than 2-fold to 1.2 mg/l culture by using Tni cells which facilitated the production of receptor on a 4 litre scale in shake cultures. The receptor was purified to homogeneity with 35% recovery giving a yield of 2 mg receptor. A further deletion at the N-terminus (betaAR 34 424/His6) eliminated proteolysis which had been observed with the original construct and also increased expression more than 5-fold to 360 pmol/mg solubilized membrane protein. This expression level is one of the highest reported for a G protein-coupled receptor (GPCR) and has enabled us to purify 10 mg betaAR for large-scale crystallization experiments. PMID- 12586388 TI - Comparison of seven different heterologous protein expression systems for the production of the serotonin transporter. AB - The rat serotonin transporter (rSERT) is an N-glycosylated integral membrane protein with 12 transmembrane regions; the N-glycans improve the ability of the SERT polypeptide chain to fold into a functional transporter, but they are not required for the transmembrane transport of serotonin per se. In order to define the best system for the expression, purification and structural analysis of serotonin transporter (SERT), we expressed SERT in Escherichia coli, Pichia pastoris, the baculovirus expression system and in four different stable mammalian cell lines. Two stable cell lines that constitutively expressed SERT (Imi270 and Coca270) were constructed using episomal plasmids in HEK293 cells expressing the EBNA-1 antigen. SERT expression in the three different inducible stable mammalian cell lines was induced either by a decrease in temperature (cell line pCytTS-SERT), the addition of tetracycline to the growth medium (cell line T REx-SERT) or by adding DMSO which caused the cells to differentiate (cell line MEL-SERT). All the mammalian cell lines expressed functional SERT, but SERT expressed in E. coli or P. pastoris was nonfunctional as assessed by 5 hydroxytryptamine uptake and inhibitor binding assays. Expression of functional SERT in the mammalian cell lines was assessed by an inhibitor binding assay; the cell lines pCytTS-SERT, Imi270 and Coca270 contained levels of functional SERT similar to that of the standard baculovirus expression system (250,000 copies per cell). The expression of SERT in induced T-REx-SERT cells was 400,000 copies per cell, but in MEL-SERT it was only 80,000 copies per cell. All the mammalian stable cell lines expressed SERT at the plasma membrane as assessed by [3H]-5 hydroxytryptamine uptake into whole cells, but the V(max) for the T-Rex-SERT cell line was 10-fold higher than any of the other cell lines. It was noticeable that the cell lines that constitutively expressed SERT grew extremely poorly, compared to the inducible cell lines whose growth rates were similar to the parental cell lines when not induced. In addition, the cell lines MEL-SERT, Imi270 and T-REx SERT all expressed fully N-glycosylated SERT and no unglycosylated inactive protein, in contrast to the baculovirus expression system where the vast majority of expressed SERT was unglycosylated and nonfunctional. PMID- 12586389 TI - PCR-based identification of Bacillus thuringiensis pesticidal crystal genes. AB - The polymerase chain reaction (PCR) is a molecular tool widely used to characterize the insecticidal bacterium Bacillus thuringiensis. This technique can be used to amplify specific DNA fragments and thus to determine the presence or absence of a target gene. The identification of B. thuringiensis toxin genes by PCR can partially predict the insecticidal activity of a given strain. PCR has proven to be a rapid and reliable method and it has largely substituted bioassays in preliminary classification of B. thuringiensis collections. In this work, we compare the largest B. thuringiensis PCR-based screenings, and we review the natural occurrence of cry genes among native strains. We also discuss the use of PCR for the identification of novel cry genes, as well as the potential of novel technologies for the characterization of B. thuringiensis strains. PMID- 12586390 TI - Photorhabdus: towards a functional genomic analysis of a symbiont and pathogen. AB - Pathogenicity and symbiosis are central to bacteria-host interactions. Although several human pathogens have been subjected to functional genomic analysis, we still understand little about bacteria-invertebrate interactions despite their ecological prevalence. Advances in our knowledge of this area are often hindered by the difficulty of isolating and working with invertebrate pathogenic bacteria and their hosts. Here we review studies on pathogenicity and symbiosis in an insect pathogenic bacterium Photorhabdus and its entomopathogenic nematode vector and model insect hosts. Whilst switching between these hosts, Photorhabdus changes from a state of symbiosis with its nematode vector to one of pathogenicity towards its new insect host and both the bacteria and the nematode then cooperatively exploit the dying insect. We examine candidate genes involved in symbiosis and pathogenicity, their secretion and expression patterns in culture and in the host, and begin to dissect the extent of their genetic coregulation. We describe the presence of several large genomic islands, putatively involved in pathogenicity or symbiosis, within the otherwise Yersinia like backbone of the Photorhabdus genome. Finally, we examine the emerging comparative genomics of the Photorhabdus group and begin to describe the interrelationship between anti-invertebrate virulence factors and those used against vertebrates. PMID- 12586391 TI - Single-strand-specific nucleases. AB - Single-strand-specific nucleases are multifunctional enzymes and widespread in distribution. Their ability to act selectively on single-stranded nucleic acids and single-stranded regions in double-stranded nucleic acids has led to their extensive application as probes for the structural determination of nucleic acids. Intracellularly, they have been implicated in recombination, repair and replication, whereas extracellular enzymes have a role in nutrition. Although more than 30 single-strand-specific nucleases from various sources have been isolated till now, only a few enzymes (S1 nuclease from Aspergillus oryzae, P1 nuclease from Penicillium citrinum and nucleases from Alteromonas espejiana, Neurospora crassa, Ustilago maydis and mung bean) have been characterized to a significant extent. Recently, some of these enzymes have been cloned, their crystal structures solved and their interactions with different substrates have been established. The detection, purification, characteristics, structure function correlations, biological role and applications of single-strand-specific nucleases are reviewed. PMID- 12586392 TI - Fluoride and organic weak acids as modulators of microbial physiology. AB - Fluoride is widely used as an anticaries agent in drinking water and a variety of other vehicles. This use has resulted in major health benefits. However, there are still open questions regarding the mechanisms of anticaries action and the importance of antimicrobial effects in caries reduction. Fluoride acts in multiple ways to affect the metabolism of cariogenic and other bacteria in the mouth. F(-)/HF can bind directly to many enzymes, for example, heme-containing enzymes or other metalloenzymes, to modulate metabolism. Fluoride is able also to form complexes with metals such as aluminum or beryllium, and the complexes, notably AlF(4)(-) and BeF(3)(-).H(2)O, can mimic phosphate with either positive or negative effects on a variety of enzymes and regulatory phosphatases. The fluoride action that appears to be most important for glycolytic inhibition at low pH in dental plaque bacteria derives from its weak-acid properties (pK(a)=3.15) and the capacity of HF to act as a transmembrane proton conductor. Since many of the actions of fluoride are related to its weak-acid character, it is reasonable to compare fluoride action to those of organic weak acids, including metabolic acids, food preservatives, non-steroidal anti-inflammatory agents and fatty acids, all of which act to de-energize the cell membrane by discharging DeltapH. Moreover, with the realization that the biofilm state is the common lifestyle for most microorganisms in nature, there is need to consider interactions of fluoride and organic weak acids with biofilm communities. Hopefully, this review will stimulate interest in the antimicrobial effects of fluoride or other weak acids and lead to more effective use of the agents for disease control and other applications. PMID- 12586393 TI - Mechanism of action of oritavancin and related glycopeptide antibiotics. AB - Oritavancin (LY333328) is a semisynthetic glycopeptide antibiotic having excellent bactericidal activity against glycopeptide-susceptible and -resistant Gram-positive bacteria. Oritavancin is the N-alkyl-p-chlorophenylbenzyl derivative of chloroeremomycin (LY264826) and is currently in phase III clinical trials for use in Gram-positive infections. Studies show that oritavancin and related alkyl glycopeptides inhibit bacterial cell wall formation by blocking the transglycosylation step in peptidoglycan biosynthesis in a substrate-dependent manner. As with other glycopeptide antibiotics, including vancomycin, the effects of oritavancin on cell wall synthesis are attributable to interactions with dipeptidyl residues of peptidoglycan precursors. Unlike vancomycin, however, oritavancin is strongly dimerized and can anchor to the cytoplasmic membrane, the latter facilitated by its alkyl side chain. Cooperative interactions derived from dimerization and membrane anchoring in situ can be of sufficient strength to enable binding to either dipeptidyl or didepsipeptidyl peptidoglycan residues of vancomycin-susceptible and -resistant enterococci, respectively. This review describes the antibacterial activity of oritavancin, and examines the evidence supporting the proposed mechanism of action for this agent and related analogs. PMID- 12586394 TI - Common domains in the initiators of DNA replication in Bacteria, Archaea and Eukarya: combined structural, functional and phylogenetic perspectives. AB - Although DNA replication is the universal process for the transmission of genetic information in all living organisms, until very recently evidence was lacking for a related structure and function in the proteins (initiators) that trigger replication in the three 'Life Domains' (Bacteria, Archaea and Eukarya). In this article new data concerning the presence of common features in the initiators of chromosomal replication in bacteria, archaea and eukaryotes are reviewed. Initiators are discussed in the light of: (i) The structure and function of their conserved ATPases Associated with various cellular Activities (AAA+) and winged helix domains. (ii) The nature of the macromolecular assemblies that they constitute at the replication origins. (iii) Their possible phylogenetic relationship, attempting to sketch the essentials of a hypothetical DNA replication initiator in the micro-organism proposed to be the ancestor of all living cells. PMID- 12586395 TI - The genetics of glycosylation in Gram-negative bacteria. AB - In recent years there has been a dramatic increase in reports of glycosylation of proteins in various Gram-negative systems including Neisseria meningitidis, Neisseria gonorrhoeae, Campylobacter jejuni, Pseudomonas aeruginosa, Escherichia coli, Caulobacter crescentus, Aeromonas caviae and Helicobacter pylori. Although this growing list contains many important pathogens (reviewed by Benz and Schmidt [Mol. Microbiol. 45 (2002) 267-276]) and the glycosylations are found on proteins important in pathogenesis such as pili, adhesins and flagella the precise role(s) of the glycosylation of these proteins remains to be determined. Furthermore, the details of the glycosylation biosynthetic process have not been determined in any of these systems. The definition of the precise role of glycosylation and the mechanism of biosynthesis will be facilitated by a detailed understanding of the genes involved. PMID- 12586396 TI - Cloning and characterization of a gene cluster for geldanamycin production in Streptomyces hygroscopicus NRRL 3602. AB - We illustrate the use of a PCR-based method by which the genomic DNA of a microorganism can be rapidly queried for the presence of type I modular polyketide synthase genes to clone and characterize, by sequence analysis and gene disruption, a major portion of the geldanamycin production gene cluster from Streptomyces hygroscopicus var. geldanus NRRL 3602. PMID- 12586397 TI - Genetic variability of the frameshift region in IS911 transposable elements from Escherichia coli clinical isolates. AB - The IS911 bacterial transposable element has been analyzed for its mechanism of transposition and for the way it controls the expression of its genes by programmed -1 translational frameshifting. In the present study the prevalence of IS911 has been determined in the Enterobacteriaceae family and in other Gram negative bacilli. Three variants, found in Escherichia coli clinical isolates and having mutations in the region implicated in frameshifting, were functionally characterized. All three were altered in their frameshifting and transposition abilities, suggesting that the frameshift region of IS911 may constitute a target for mutations reducing the transposition frequency of this mobile element in natural populations of E. coli. PMID- 12586398 TI - A variant of the hyperthermophile Archaeoglobus fulgidus adapted to grow at high salinity. AB - A variant of Archaeoglobus fulgidus VC-16 was isolated from cultures obtained after a stepwise transfer from media containing 1.8-6.3% NaCl by a plating independent, selected-cell cultivation technique, using a laser microscope. This variant, A. fulgidus VC-16S, had a higher growth rate throughout the salt range of the parental strain, but was also able to grow in media containing NaCl up to 6.3%, whereas the parental strain could not grow above 4.5% NaCl. Diglycerol phosphate (DGP), only encountered in the Archaeoglobales, was the major solute accumulated under supra-optimal salinities, whereas at supra-optimal growth temperatures di-myo-inositol phosphate was the predominant solute. The accumulation of compatible solutes during growth of variant VC-16S was lower than in the parental strain within 1.8-4.5% NaCl, but the levels of compatible solutes, including DGP, increased sharply in the variant at higher salinities (5.5 and 6.0%). This variant represents, at this time, one of the most halophilic hyperthermophiles known, and its ability to grow at high salinity appears to be due to the massive accumulation of DGP. PMID- 12586399 TI - Lactate carbon does not enter the sugars of lipopolysaccharide when gonococci are grown in a medium containing glucose and lactate: implications in vivo. AB - In media containing glucose, lactate stimulates the metabolism of gonococci at concentrations that simulate conditions in vivo. Nuclear magnetic resonance (NMR) spectroscopy of (13)C-labelled lipids obtained from gonococci grown in a synthetic medium with (13)C-labelled lactate and unlabelled glucose (culture A), (13)C-labelled glucose alone (culture B) or (13)C-labelled glucose and unlabelled lactate (culture C) showed lactate carbon was not present in glycerol/ethanolamine residues of lipids from culture A. This indicated that, in the presence of glucose, lactate gluconeogenesis is shut down. Hence, the stimulation of metabolism could result from the production of extra energy because lactate is used solely for conversion to acetyl-CoA, the precursor of fatty acid synthesis and the components of the tricarboxylic acid cycle. In this paper, additional evidence for lack of gluconeogenesis has been sought using a different approach. The carbohydrate moieties of lipopolysaccharide (LPS) have been examined for lactate carbon after gonococci were grown with lactate and glucose. Two methods were used: NMR spectroscopy of (13)C-labelled lipopolysaccharide purified from the three cultures described above showed that, in the presence of glucose, lactate carbon, in contrast to glucose carbon, was not in the carbohydrate moiety. Also, (14)C-labelled lactate was added to a culture containing unlabelled glucose and lactate (culture A) and [(14)C]glucose to cultures containing unlabelled glucose without unlabelled lactate (culture B) and with unlabelled lactate (culture C). When LPS samples purified from these cultures were subjected to hydrazinolysis, the ratio of the radioactivity of water-soluble products (carbohydrate moieties) to those of chloroform-soluble products (fatty acids) was much lower when [(14)C]lactate was used in culture A, than when [(14)C]glucose was used in cultures B and C. Thus, in the presence of glucose, lactate carbon, unlike glucose carbon, is incorporated predominantly into fatty acids of LPS, not into its carbohydrate moieties. There is no doubt, therefore, that gluconeogenesis is shut off when lactate is present with glucose and there is a consequent stimulation of metabolism. This probably occurs in vivo on mucous surfaces, where gonococci are surrounded by a mixture of glucose and lactate in the secretions. PMID- 12586400 TI - Deacetoxycephalosporin C synthase isozymes exhibit diverse catalytic activity and substrate specificity. AB - The biosynthesis of cephalosporins involving a thiozolidine ring expansion is catalyzed by deacetoxycephalosporin C synthase (DAOCS). In this study, three DAOCS isozymes were cloned and expressed as active enzymes together with Streptomyces jumonjinensis DAOCS that was newly isolated and partially characterized. The enzymes showed excellent substrate conversion for penicillin G, phenethicillin, ampicillin and carbenicillin, but they were less effective in the ring expansion of penicillin V, amoxicillin and metampicillin. Streptomyces clavuligerus DAOCS was the most active among the recombinant enzymes. The results also showed that truncation of 20 amino acids at the C-terminus of the Acremonium chrysogenum deacetoxy/deacetylcephalosporin C synthase polypeptide did not affect penicillin ring expansion. PMID- 12586401 TI - Expression of the ech42 (endochitinase) gene of Trichoderma atroviride under carbon starvation is antagonized via a BrlA-like cis-acting element. AB - Expression of the endochitinase encoding ech42 gene of the mycoparasite Trichoderma atroviride is subject to control by several environmental signals, including derepression by carbon starvation. In order to identify promoter areas involved in control by this condition, we prepared fusions of several mutant forms of the ech42 promoter to the Aspergillus niger goxA gene as a reporter. Removal of a 130-bp fragment comprising a binding site for the carbon catabolite repressor Cre1, an AGGGG element and three separate binding sites identical and highly similar, respectively, to those for the Aspergillus nidulans regulator of conidiation BrlA resulted in a three-fold increase in derepression under carbon starvation. A truncation of the promoter to 196 bp, which removed all of the observed DNA binding motifs, resulted in five-fold derepression. In vitro protein DNA binding analyses showed that only the BrlA-like sites, but neither the AGGGG element nor the Cre1 binding site, bound proteins from cell-free extracts from carbon-starved mycelia of T. atroviride. Thus this study identifies a new regulator of chitinase gene expression in Trichoderma, a BrlA-like binding motif. PMID- 12586402 TI - Molecular cloning of the gene for 2,6-beta-D-fructan 6-levanbiohydrolase from Streptomyces exfoliatus F3-2. AB - The gene encoding a 2,6-beta-D-fructan 6-levanbiohydrolase (LF2ase) (EC 3.2.1.64) that converts levan into levanbiose was cloned from the genomic DNA of Streptomyces exfoliatus F3-2. The gene encoded a signal peptide of 37 amino acids and a mature protein of 482 amino acids with a total length of 1560 bp and was successfully expressed in Escherichia coli. The similarities of primary structure were observed with levanases from Clostridium acetobutylicum, Bacillus subtilis, B. stearothermophilus (51.0-54.3%) and with LF2ase from Microbacterium levaniformans (53.9%). The enzyme from S. exfoliatus F3-2 shared the conserved six domains and the completely conserved five amino acid residues with family 32 glycosyl hydrolases, which include levanase, inulinase, and invertase. These observations led to the conclusion that the enzyme belongs to family 32 glycosyl hydrolases. PMID- 12586403 TI - Growth promotion of the edible fungus Pleurotus ostreatus by fluorescent pseudomonads. AB - Bacteria were isolated from the mycelial surface of Pleurotus ostreatus and their role in fruiting body induction (fructification) of the edible mushroom P. ostreatus was investigated. Analysis of the bacterial community that colonized the mycelium showed that the species composition and numbers of culturable bacteria differed according to the developmental stage of P. ostreatus. In particular, the population size of fluorescent pseudomonads increased during fruiting body induction. An experiment showed that inoculation of pure cultures of the mycelium with strains of fluorescent Pseudomonas spp. isolated from the mycelial plane of commercially produced mushrooms promoted the formation of primordia and enhanced the development of the basidiome of P. ostreatus. Results of this research strongly suggest that inoculation of the mycelium with specific bacteria may have beneficial applications for mushroom production. PMID- 12586404 TI - Vacuolar system of ungerminated Colletotrichum graminicola conidia: convergence of autophagic and endocytic pathways. AB - Vacuoles of ungerminated Colletotrichum graminicola conidia engulf cytoplasmic structures by a process analogous to microautophagy, demonstrated by using a vacuolar membrane acid phosphatase marker. Fusion of vesicles with vacuoles, without deposition of the acid phosphatase reaction product has been observed, suggesting other pathways of material delivery to vacuoles than microautophagy. Plasma membrane invaginations, multivesicular bodies and retention of neutral red into small vesicles, which were internalized by the vacuole, were verified. These results provided evidence for endocytosis and an active endosomal system. Together, our findings with C. graminicola demonstrated that vacuoles are very dynamic compartments, playing roles in autophagy and endocytic processes. PMID- 12586405 TI - Cloning, expression and evolution of the gene encoding the elongation factor 1alpha from a low thermophilic Sulfolobus solfataricus strain. AB - The gene encoding the elongation factor 1alpha (EF-1alpha) from the archaeon Sulfolobus solfataricus strain MT3 (optimum growth temperature 75 degrees C) was cloned, sequenced and expressed in Escherichia coli. The structural and biochemical properties of the purified enzyme were compared to those of EF-1alpha isolated from S. solfataricus strain MT4 (optimum growth temperature 87 degrees C). Only one amino acid change (Val15-->Ile) was found. Interestingly, the difference was in the first guanine nucleotide binding consensus sequence G(13)HIDHGK and was responsible for a reduced efficiency in protein synthesis, which was accompanied by an increased affinity for both guanosine diphosphate (GDP) and guanosine triphosphate (GTP), and an increased efficiency in the intrinsic GTPase activity. Despite the different thermophilicities of the two microorganisms, only very marginal effects on the thermal properties of the enzyme were observed. Molecular evolution among EF-1alpha genes from Sulfolobus species showed that the average rate of nucleotide substitution per site per year (0.0312x10(-9)) is lower than that reported for other functional genes. PMID- 12586406 TI - Role of Saccharomyces cerevisiae serine O-acetyltransferase in cysteine biosynthesis. AB - Some strains of Saccharomyces cerevisiae have detectable activities of L-serine O acetyltransferase (SATase) and O-acetyl-L-serine/O-acetyl-L-homoserine sulfhydrylase (OAS/OAH-SHLase), but synthesize L-cysteine exclusively via cystathionine by cystathionine beta-synthase and cystathionine gamma-lyase. To untangle this peculiar feature in sulfur metabolism, we introduced Escherichia coli genes encoding SATase and OAS-SHLase into S. cerevisiae L-cysteine auxotrophs. While the cells expressing SATase grew on medium lacking L-cysteine, those expressing OAS-SHLase did not grow at all. The cells expressing both enzymes grew very well without L-cysteine. These results indicate that S. cerevisiae SATase cannot support L-cysteine biosynthesis and that S. cerevisiae OAS/OAH-SHLase produces L-cysteine if enough OAS is provided by E. coli SATase. It appears as if S. cerevisiae SATase does not possess a metabolic role in vivo either because of very low activity or localization. For example, S. cerevisiae SATase may be localized in the nucleus, thus controlling the level of OAS required for regulation of sulfate assimilation, but playing no role in the direct synthesis of L-cysteine. PMID- 12586407 TI - The Bacillus subtilis transition state regulator AbrB binds to the -35 promoter region of comK. AB - Genetic competence is a differentiation process initiated by Bacillus subtilis as a result of nutritional deprivation, and is controlled by a complex signal transduction cascade. The promoter of comK, encoding the competence transcription factor, is regulated by at least four different transcription factors: Rok, CodY, DegU and ComK itself. Genetic data have shown that comK expression is influenced by the transition state regulator AbrB as well. In this paper we show that AbrB binds specifically to the comK promoter and covers the RNA polymerase binding site, making it the fifth transcription factor regulating the activity of the comK promoter. PMID- 12586408 TI - Molecular and biochemical characterization of mechanosensitive channels in Corynebacterium glutamicum. AB - Database searches in the Corynebacterium glutamicum genome sequence revealed homologs of the mechanosensitive channels MscL and YggB of Escherichia coli. To elucidate the physiological role of these putative channels deletion mutants were constructed. Betaine efflux induced by osmotic downshock of the mscL deletion mutant was nearly identical to that of the wild-type, whereas the yggB deletion mutant showed a reduced efflux rate. Interestingly, the double deletion strain, which was expected to have an even more decreased capability of betaine excretion, had only a slightly reduced efflux rate compared to the wild-type and did not show an increased mortality after osmotic downshift. These results led to the hypothesis that C. glutamicum may possess a third type of mechanosensitive channel not related to the MscL and YggB/KefA families. Furthermore it is unlikely that an MscM-like activity is responsible for the betaine efflux, because of the high transport capacity detected in the double deletion mutant. PMID- 12586409 TI - Human intestinal tissue tropism of intimin epsilon O103 Escherichia coli. AB - Human intestinal in vitro organ culture was used to assess the tissue tropism of human isolates of Escherichia coli O103:H2 and O103:H- that express intimin epsilon. Both strains showed tropism for follicle associated epithelium and limited adhesion to other regions of the small and large intestine. This is similar to the tissue tropism shown by intimin gamma enterohaemorrhagic (EHEC) O157:H7, but distinct from that of intimin alpha enteropathogenic (EPEC) O127:H6. PMID- 12586410 TI - Impala, a transposon from Fusarium oxysporum, is active in the genome of Penicillium griseoroseum. AB - An autonomous impala transposon trapped in Fusarium oxysporum by insertion within the niaD gene encoding nitrate reductase was introduced in the genome of the fungus Penicillium griseoroseum, a producer of pectinase enzymes. Through a phenotypic assay, we demonstrate that this element is able to excise from the niaD gene and to reinsert at new genomic positions. As in the original host, impala inserts into a TA site and footprints left by impala excisions are generally 5 bp. The fact that impala is able to transpose in P. griseoroseum offers the opportunity to develop a gene-tagging system based on this element with the objective to detect and clone genes related in pectinase production. PMID- 12586411 TI - The wbiA locus is required for the 2-O-acetylation of lipopolysaccharides expressed by Burkholderia pseudomallei and Burkholderia thailandensis. AB - Burkholderia pseudomallei and Burkholderia thailandensis express similar O antigens (O-PS II) in which their 6-deoxy-alpha-L-talopyranosyl (L-6dTalp) residues are variably substituted with O-acetyl groups at the O-2 or O-4 positions. In previous studies we demonstrated that the protective monoclonal antibody, Pp-PS-W, reacted with O-PS II expressed by wild-type B. pseudomallei strains but not by a B. pseudomallei wbiA null mutant. In the present study we demonstrate that WbiA activity is required for the acetylation of the L-6dTalp residues at the O-2 position and that structural modification of O-PS II molecules at this site is critical for recognition by Pp-PS-W. PMID- 12586412 TI - Development of a PCR-based assay for rapid and reliable identification of pathogenic Fusaria. AB - Identification of Fusarium species has always been difficult due to confusing phenotypic classification systems. We have developed a fluorescent-based polymerase chain reaction assay that allows for rapid and reliable identification of five toxigenic and pathogenic Fusarium species. The species includes Fusarium avenaceum, F. culmorum, F. equiseti, F. oxysporum and F. sambucinum. The method is based on the PCR amplification of species-specific DNA fragments using fluorescent oligonucleotide primers, which were designed based on sequence divergence within the internal transcribed spacer region of nuclear ribosomal DNA. Besides providing an accurate, reliable, and quick diagnosis of these Fusaria, another advantage with this method is that it reduces the potential for exposure to carcinogenic chemicals as it substitutes the use of fluorescent dyes in place of ethidium bromide. Apart from its multidisciplinary importance and usefulness, it also obviates the need for gel electrophoresis. PMID- 12586413 TI - Phenotypic evidence for inducible multiple antimicrobial resistance in Salmonella choleraesuis. AB - Multiple antimicrobial resistance (MAR) in Salmonella choleraesuis is becoming a major concern. It has been demonstrated that a MAR phenotype can be induced in Escherichia coli and other members of the Enterobacteriaceae by exposing the isolates to salicylates, various antimicrobials, or organic solvents used to combat and control bacterial infection. Therefore the purpose of the present study was to determine whether this marA-associated MAR-phenotype is inducible in S. choleraesuis. Isolates used in the present study were able to withstand toxic effects of the organic solvent cyclohexane naturally, or following exposure to the inducing compounds salicylate, tetracycline, or chloramphenicol. All isolates possessed fragments of marA with the predicted size of 408 bp when amplified using marA-specific primers by PCR. The resulting PCR products that were sequenced revealed that amplified S. choleraesuis marA was 99% and 85% homologous to the published Salmonella typhimurium and E. coli marA sequences respectively. Minimum inhibitory concentrations of tetracycline (P<0.08), chloramphenicol (P<0.001), rifampin (P<0.08), and nalidixic acid (P<0.001) against cyclohexane tolerant mutants were significantly increased when compared with wild-type S. choleraesuis. Northern hybridization signals for both marA and acrB were increased in the induced isolates when compared to uninduced controls while soxS expression did not change between induced and uninduced cultures. The results suggest that marA is present in S. choleraesuis and a MAR-phenotype is inducible in S. choleraesuis presumably due to the overexpression of marA and acrB and not to the overexpression of soxS. PMID- 12586414 TI - CANON and Anammox in a gas-lift reactor. AB - Anoxic ammonium oxidation (Anammox) and Completely Autotrophic Nitrogen removal Over Nitrite (CANON) are new and promising microbial processes to remove ammonia from wastewaters characterized by a low content of organic materials. These two processes were investigated on their feasibility and performance in a gas-lift reactor. The Anammox as well as the CANON process could be maintained easily in a gas-lift reactor, and very high N-conversion rates were achieved. An N-removal rate of 8.9 kg N (m(3) reactor)(-1) day(-1) was achieved for the Anammox process in a gas-lift reactor. N-removal rates of up to 1.5 kg N (m(3) reactor)(-1) day( 1) were achieved when the CANON process was operated. This removal rate was 20 times higher compared to the removal rates achieved in the laboratory previously. Fluorescence in situ hybridization showed that the biomass consisted of bacteria reacting to NEU, a 16S rRNA targeted probe specific for halotolerant and halophilic Nitrosomonads, and of bacteria reacting to Amx820, specific for planctomycetes capable of Anammox. PMID- 12586415 TI - Involvement of a quinoprotein (PQQ-containing) alcohol dehydrogenase in the degradation of polypropylene glycols by the bacterium Stenotrophomonas maltophilia. AB - Previous work has shown that when the bacterium Stenotrophomonas maltophilia is grown on polypropylene glycol, different dye-linked polypropylene glycol dehydrogenase (PPG-DH) activities are induced during growth. Here the purification and characterization of the dehydrogenase activity induced in the stationary phase, and present in the periplasmic space, is described. The homogeneous enzyme preparation obtained consists of a homodimeric protein with a molecular mass of about 123 kDa and an isoelectric point of 5.9. The cofactor of the enzyme appeared to be pyrroloquinoline quinone (PQQ), no heme c was present, and holo-enzyme contained two PQQ molecules per enzyme molecule. In these respects, PPG-DH described here is similar to already known quinoprotein alcohol dehydrogenases, but in other respects, it is different. Therefore, it is suggested that PPG-DH could be a new type of quinoprotein alcohol dehydrogenase. Based on its strong preference for polyols, PPG-DH seems well fitted to carry out the first step in the degradation of PPGs, synthetic polymers containing a variety of hydroxyl groups. PMID- 12586416 TI - Cyanobacterial leader peptides for protein secretion. AB - The leader peptide of the major secreted protein PilA1 of the cyanobacterium Synechocystis sp. strain PCC 6803 and several artificial leader peptides have been used to study secretion of the reporter protein lichenase to the culture medium. The strains of Synechocystis carrying lichenase with the leader sequences of PilA and with the leader sequence of Slr2016 efficiently secreted the reporter protein. The artificial leader sequence that was characterized by the overall positive charge (as PilA1 and Slr2016 leaders) also allowed secretion. The artificial leader with negative charge, however, did not allow secretion of the reporter protein. Moreover, no secreted proteins have been isolated from this strain using conventional techniques for preparation of secreted proteins. These data suggest that the general secretion pathway in cyanobacteria, at least for pilins, recognizes the overall charge of the leader sequences, and operates in a sequence-non-specific manner. PMID- 12586417 TI - Evidence for limited species diversity of bacteriochlorophyll b-containing purple nonsulfur anoxygenic phototrophs in freshwater habitats. AB - Thirteen new isolates of bacteriochlorophyll b-containing purple nonsulfur bacteria were isolated from four freshwater habitats using specific enrichment methods including the use of long wavelength filters and extincting dilution of the inoculum. The new isolates were compared with the type strain of Blastochloris viridis, strain DSM 133(T), as regards pigments, morphology, carbon nutrition, and phylogeny. All new isolates were budding bacteria, and phototrophic mass cultures were green, brown, or brown-green in color. The pattern of carbon sources photocatabolized were similar in all strains; however, sugars, both mono- and disaccharides, were widely used by the new isolates while they did not support growth of strain DSM 133(T). Phylogenetic analysis showed all new strains to cluster tightly with the type strain with the exception of one brown-colored strain and a mildly thermophilic strain. The results suggest that in contrast to purple nonsulfur bacteria containing bacteriochlorophyll a, those containing bacteriochlorophyll b may not be morphologically or phylogenetically diverse, and group into a tight phylogenetic clade distinct from all other anoxygenic phototrophs. PMID- 12586418 TI - The regulatory elements of the Mycobacterium tuberculosis gene Rv3881c function efficiently in Escherichia coli. AB - We report efficient expression of the Mycobacterium tuberculosis gene Rv3881c in Escherichia coli from its M. tuberculosis promoter, attributable to an E. coli consensus Pribnow box and ribosome binding site. The N-terminal sequence of the recombinant E. coli-generated protein was identical to the predicted open reading frame of Rv3881c and transcription of the Rv3881c gene initiated at the same nucleotide position in both bacteria. We demonstrate the utility of this promoter for rapid analysis of expression in E. coli of heterologous gene constructs, for subsequent expression from the genomes of slow-growing mycobacteria such as Mycobacterium bovis-BCG. M. tuberculosis Rv3881c homologues were present in other pathogenic mycobacteria such as M. bovis-BCG, Mycobacterium szulgai and Mycobacterium kansasii. PMID- 12586419 TI - Siderophore production by the magnetic bacterium Magnetospirillum magneticum AMB 1. AB - Siderophore production by the magnetic bacterium Magnetospirillum magneticum AMB 1 is elicited by sufficient iron rather than by iron starvation. In order to clarify this unusual pattern, siderophore production was monitored in parallel to iron assimilation using the chrome azurol sulfonate assay and the ferrozine method respectively. Iron concentration lowered approximately five times less than its initial concentration only within 4 h post-inoculation, rendering the medium iron deficient. A concentration of at least 6 microM Fe(3+) is required to initiate siderophore production. The propensity of M. magneticum AMB-1 for the assimilation of large amounts of iron accounts for the rapid depletion of iron in the medium, thereby triggering siderophore excretion. M. magneticum AMB-1 produces both hydroxamate and catechol siderophores. PMID- 12586420 TI - The effect of cell density and specific growth rate on accessory gene regulator and toxic shock syndrome toxin-1 gene expression in Staphylococcus aureus. AB - A continuous culture technique utilising a variant of Staphylococcus aureus 8325 4 containing transcriptional gene fusions was used to investigate the relationships between cell density (OD(600)), steady-state specific growth rate (mu) and expression of both agr (accessory gene regulator) and tst (toxic shock syndrome toxin-1). The expression of these genes was assessed by two single-copy independently arranged chromosomal-based reporter systems, beta-galactosidase agr P3 promoter fusion and a lux-tst promoter fusion. Cell density and specific agr expression were found to be positively correlated. In the model, the minimum cell density predicted to promote specific agr expression was an OD(600) of 0.14, equivalent to 1.2x10(8) CFU ml(-1). No direct relationship between cell density and specific tst expression was detected. Specific expressions of agr and tst were not correlated with specific growth rate and there appeared to be no direct link between agr and tst specific expression. The results support the hypothesis that agr is a functional unit of quorum sensing and that the amount of specific expression of tst is modulated independently of agr. PMID- 12586421 TI - Regulation of the flavorubredoxin nitric oxide reductase gene in Escherichia coli: nitrate repression, nitrite induction, and possible post-transcription control. AB - Escherichia coli flavorubredoxin is a new type of cytoplasmic nitric oxide (NO) reductase, which shows NO reductase activity within the range of the canonical membrane-bound heme b(3)-iron NO reductases. Using reverse-transcription polymerase chain reaction we show that although the flavorubredoxin gene (flrd) is transcribed in both aerobic and anaerobic conditions, anaerobiosis induced transcription up to 12-fold, under fermentative conditions; a 28-fold stimulation was observed in an E. coli fnr mutant strain, showing that the flavorubredoxin gene is negatively regulated by FNR. The level of anaerobic transcription was repressed three-fold by nitrate, but induced 47-fold by nitrite. The transcription factors NarL and NarP are not essential for flrd expression. Furthermore, the addition of NO within the physiological range of concentrations does not induce anaerobic transcription of flrd. Since two other E. coli proteins are known to exhibit NO reductase activity, flavohemoglobin and the pentaheme cytochrome c nitrite reductase, we have also compared the concentrations of their mRNAs with those of flavorubredoxin, under the same growth conditions. Transcription of the putative transcriptional activator of flavorubredoxin, ygaA, is also regulated by the absence of oxygen and the presence of nitrite. Levels of FlRd protein did not correlate with mRNA levels. The results reveal that a complex regulation of flavorubredoxin expression is operative, possibly by both transcriptional and post-transcriptional mechanisms. PMID- 12586422 TI - Anosmin-1 immunoreactivity during embryogenesis in a primitive eutherian mammal. AB - Kallmann syndrome is hypogonadotropic hypogonadism coupled with anosmia. A morphological study found that the endocrine disorder in X-linked Kallmann syndrome is due to failed migration of gonadotropin releasing-hormone (GnRH) neurons from the olfactory placode to the brain during development. Anosmia results from agenesis of the olfactory bulbs and tracts. The gene responsible for the X-linked form of Kallmann syndrome, KAL-1, has been characterized. The orthologues of KAL-1 have been isolated in the chick and the zebrafish, but still await identification in rodents. In the present study, we used polyclonal and monoclonal antibodies to the human KAL-1 encoded protein, anosmin-1, in a primitive mammal, the Asian musk shrew. Musk shrews are insectivores and are therefore evolutionarily closer to primates than rodents. By immunoblot analysis of musk shrew tissues, a band of the expected apparent molecular mass (95 kDa) was detected in several structures of the central nervous system, but not in liver or muscle, which is consistent with the gene expression pattern previously reported in the chick. By immunohistochemical analysis, anosmin-1 was detected in the developing olfactory epithelium, the olfactory, vomeronasal and terminalis nerves, the olfactory bulbs, the cerebellum and the cerebral cortex and in several other regions of the brain, during musk shrew embryogenesis. Furthermore, migrating gonadotropin releasing-hormone (GnRH)-immunoreactive neurons were seen in close association with anosmin-1-immunoreactive fibers. Assuming that the protein is present at the surface of these fibers, we suggest a possible direct role of anosmin-1 in the migration of GnRH neurons in this species. PMID- 12586424 TI - Integrins are involved in synaptogenesis, cell spreading, and adhesion in the postnatal brain. AB - Integrins are a major family of heterodimeric surface glycoproteins that act as adhesion molecules, have a spectrum of extracellular matrix (ECM) molecules as their ligands, and regulate a variety of cellular functions. Integrins are known to be critical to embryonic brain development, and recent studies have indicated their essential role in adult brain function, although their role in postnatal brain development and function has not been examined. Here, we used the organotypic slice culture system to investigate the role of integrins in postnatal hippocampal development by exposing the tissue to either an integrin competitive antagonist, the peptide GRGDSP containing Arg-Gly-Asp (RGD) attachment site, or to function-blocking beta(1)-integrin antibodies to disrupt integrin interactions. These experiments revealed that beta(1)-integrin antibodies interfered with spreading of the culture, resulting in a rapid and marked diminution of slice area. beta(1)-integrin antibodies and RGD peptide disrupted cell adhesion, causing cell detachment and migration of glial cells from the explant. The majority of the detached cells were of macroglial origin and switched to expression of the intermediate filament proteins vimentin and nestin, suggesting a developmental regression. The organotypic organization of slice cultures was not affected, although exposure to either integrin antagonist or antibody resulted in a statistically significant reduction in the number of synapses measured in the apical dendrites of CA1 pyramidal neurons. The results demonstrate that integrins markedly affect postnatal CNS development, in both ultrastructural construction and organizational processes. PMID- 12586423 TI - Photoreceptor-specific overexpression of platelet-derived growth factor induces proliferation of endothelial cells, pericytes, and glial cells and aberrant vascular development: an ultrastructural and immunocytochemical study. AB - Platelet-derived growth factor (PDGF) is necessary for the normal development of the retinal vasculature and its overexpression is likely to contribute to proliferative retinal disorders, such as proliferative vitreoretinopathy. Transgenic mice that overexpress PDGF-B in the photoreceptors (rho/PDGF-B mice) develop traction retinal detachment. In the present study, a detailed histopathological analysis was performed in rho/PDGF-B mice. In these transgenic mice, endothelial cells, pericytes, and glial cells begin to proliferate at postnatal day 7 (P7). All three cell types increase in numbers, forming a highly vascularized cell mass, which reaches a maximum thickness at P14. Cords of endothelial cells and glia invade the retina and exert traction, generating retinal folds; however, the deep capillary bed never forms. Griffonia simplicifolia isolectin B4 (GSA)-positive endothelial cells form tubes and penetrate the retina to the level of the outer plexiform layer, but they never interconnect to form the deep capillary bed. The vessels within the cell mass are patent, but have a very immature morphology. They often are thin-walled with fenestrations. Pericytes and glial cells are usually found in clusters and are not associated with the abnormal vessels. The lack of this association may account for the failure to form a mature vasculature. PMID- 12586425 TI - Localization of ApoER2, VLDLR and Dab1 in radial glia: groundwork for a new model of reelin action during cortical development. AB - The reelin signaling pathway regulates laminar positioning of radially migrating neurons during cortical development. It has been suggested that reelin secreted by Cajal-Retzius cells in the marginal zone could provide either a stop or an attractant signal for migratory neurons expressing reelin receptors, but the proposed models fail to explain recent experimental findings. Here we provide evidence that the reelin receptor machinery, including the lipoprotein receptors ApoER2 and VLDLR along with the cytoplasmic adaptor protein Dab1, is located in radial glia precursors whose processes span the entire cortical wall from the ventricular zone to the pial surface. Moreover, in reeler mice, defective in reelin, decreased levels of Dab1 in the ventricular zone correspond to an accumulation of the protein in radial end-feet beneath the pia matter. Our results support that neural stem cells receive a functional reelin signal. They are also consistent with a working model of reelin action, according to which reelin signaling on the newborn neuron-inherited radial process regulates perikaryal translocation and positioning. PMID- 12586426 TI - Disturbance of oligodendrocyte development, hypomyelination and white matter injury in the neonatal rat brain after intracerebral injection of lipopolysaccharide. AB - Increasing data provide support for the hypothesis that brain inflammation plays an important role in injury to developing white matter. In the present study, inflammatory responses in the neonatal rat brain were investigated following lipopolysaccharide (LPS) administration at postnatal day 5. LPS-induced brain injury was examined in brain sections 24 h, 3 and 9 days after LPS injection. White matter rarefaction was observed in 50% of the rat brains (three out of six) 24 h after LPS injection. Lateral ventricle enlargement was found in 100% (four out of four) and 89% (eight out of nine) of rat brains 3 and 9 days after LPS administration, respectively. White matter necrosis was found in three out of nine brains injected with LPS on P14. None of these injuries was observed in any control rat brains. No histological changes in gray matter were noted in the LPS injected rat brain. Proinflammatory cytokines, tumor necrosis factor-alpha (TNFalpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS) in the rat brain were greatly induced after LPS administration. Activated astrocytes and microglia/macrophages were found in the affected rat brains. Double-labeling showed that IL-1beta and iNOS expressing cells were microglia/macrophages. Injury to or delayed development of immature oligodendrocytes (OLs) was evident by decreased immunostaining for both O4 and O1 antibodies, markers for developing immature OLs, in the LPS-injected as compared to the control rat brain. LPS also resulted in hypomyelination, as indicated by reduced myelin basic protein (MBP) immunostaining in the P8 rat brain. Co administration of IL-1 receptor antagonist (IL-1Ra) with LPS reduced brain injury by improving myelination and subsequent reduction of lateral ventricle enlargement. These results indicate that developing OLs may be the target cells for LPS-induced brain injury and inflammatory cytokines are possible mediators of LPS-induced brain injury. PMID- 12586427 TI - The effect of a global birth asphyxia on the ontogeny of BDNF and NGF protein expression in the juvenile brain. AB - Neurotrophic growth factors are strongly upregulated following brain injury in order to limit the amount of delayed apoptotic cell death. In particular, the neurotrophins NGF and BDNF are upregulated following injury and offer neuroprotection when administered after brain injury. Further, both growth factors are involved in the control of neural proliferation and plasticity during both development and recovery from injury. We used a model of global birth asphyxia in the rat to follow the ontogeny of BDNF and NGF protein levels within the normal and asphyctic hippocampus and cerebellum for the first 28 days of postnatal life. In contrast to what is seen in the injured adult brain, we see an early and long lasting decrease in NGF content within the asphyctic hippocampus, whereas cerebellar NGF content showed a delayed increase following asphyxia. Asphyxia also caused a delayed increase in BDNF content within the hippocampus but decreased BDNF levels within the cerebellum. Further, a comparison of the ontogeny of plasma corticosterone over development shows that endogenous BDNF protein levels are not sensitive to the dramatic increase in circulating corticosterone that occurs at the end of the stress hyporesponsive period. In summary, we find that perinatal birth asphyxia causes opposing changes in NGF and BDNF protein expression in a spatio-temporal-dependent manner. These results point to the need for more detailed studies on the mechanisms of action of BDNF and NGF within the developing brain before these can be used therapeutically following birth asphyxia in man. PMID- 12586428 TI - Partial segregation of posterior crista and saccular fibers to the nodulus and uvula of the cerebellum in mice, and its development. AB - The projection of the posterior canal crista and saccular afferents to the cerebellum of embryonic and neonatal mice was investigated using carbocyanine dyes. Anterograde tracing from these two endorgans reveals a partial segregation of these two sets of afferents. The saccule projects predominantly to the uvula, with very minor input to the nodulus. The posterior canal projects mainly to the nodulus and, to a lesser extent, to the uvula. Retrograde tracing from the uvula and nodulus confirms this partial segregation for these two endorgans and extends it to other vestibular endorgans. Uvular injections result in many more labeled fibers in the gravistatic maculae than in the canals' cristae. In contrast, nodular injection reveals many more labeled fibers in the canal cristae than in the gravistatic maculae. This partial segregation may play a role in the information processing in these folia. Our developmental data suggest that the initial segregation at E17 coincides with the formation of the postero-lateral fissure. This embryonic segregation of the primary vestibular mossy fibers to the uvula and nodulus commences long before the maturity of their targets, the granule cells and unipolar brush cells. Thus, the segregation of the primary vestibular projection to the uvula and nodulus does not depend on cues related to the target cells. Rather, the segregation may reflect more global cerebellar patterning mechanisms involving guidance for the vestibular afferent fibers independent of the future target cells. PMID- 12586429 TI - Effects of ethanol on neurotrophic factors, apoptosis-related proteins, endogenous antioxidants, and reactive oxygen species in neonatal striatum: relationship to periods of vulnerability. AB - The developing central nervous system is extremely sensitive to ethanol, with well-defined temporal periods of vulnerability. Many brain regions are particularly susceptible to ethanol during the early neonatal period, corresponding to the human third trimester, which represents a dynamic period of growth and differentiation. For this study, neonatal rats were acutely exposed to ethanol or control conditions at a neonatal age when the developing striatum has been shown to be vulnerable to ethanol (postnatal day 3 [P3]), and at a later age (P14), when this developing region is relatively ethanol-resistant. We then analyzed basal levels of neurotrophic factors (NTFs), and ethanol-mediated changes in NTFs, apoptosis-related proteins, antioxidants, and reactive oxygen species (ROS) generation, which may underlie this differential temporal vulnerability. Sequential analyses were made following ethanol exposure on these two postnatal days, with assessments of NTFs nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT 4); apoptosis-related proteins Bcl-2, Bcl-xl, Bax, Akt and c-jun N-terminal kinase (JNK); antioxidants superoxide dismutase, glutathione reductase and catalase; and ROS. The results indicated that basal levels of BDNF, and to some degree NGF, were greater at the older age, and that ethanol exposure at the earlier age elicited considerably more pro-apoptotic and fewer pro-survival changes than those produced at the later age. Thus, differential temporal vulnerability to ethanol in this CNS region appears to be related to differences in both differential levels of protective substances (e.g. NTFs), and differential cellular responsiveness which favors apoptosis at the most sensitive age and survival at the resistant age. PMID- 12586430 TI - Repeated neonatal pain influences maternal behavior, but not stress responsiveness in rat offspring. AB - Early preterm neonates in the Neonatal Intensive Care Unit (NICU) are subjected to repeated painful procedures which could sensitize their responses to pain and potentiate neuroendocrine and behavioral responses to subsequent stressors in the long-term. In this study, we used the model of the neonatal rat to test the effects of repeated pain during the first 2 weeks of life on neuroendocrine responses (CRF, ACTH and corticosterone) to stressors varying in intensity and on maternal behavior in the postnatal period. To closely mimic the type of repeated painful stimulus experienced by preterm neonates (i.e., heelstick), neonatal rats aged day 2-14 were submitted daily to having their rear heels warmed to 34 degrees C and pricked (handled and pain, HP) or not (handled, H) with a needle. For the procedure, all pups were separated from their mothers for a total period of 15 min and reunited afterwards. Unhandled (UH) pups not subjected to daily maternal separation were used as controls. On days 6 and 12, litters from the HP and H groups were videotaped for 90 min upon return with the mother and maternal behavior was analyzed. Frequency of ultrasonic vocalizations (USV) were recorded during the procedure and upon return of pups with the mother. On day 15 and 20, rat pups from all groups were exposed to a 3-min ether vapor stressor or to an openfield for 10 min. Plasma ACTH and corticosterone concentrations were determined at 0, 5, 30, 60 and 120 min after stress onset. Our results show that repeated pain did not modify body weight of the pups, however, on day 6 of life, maternal pup grooming was increased significantly (P<0.05) in the HP group compared to the H group. Frequency of USV was not changed between H and HP rats either during the separation or after reunion with the mother. Plasma ACTH and corticosterone levels under basal or stimulated conditions were not different between UH, H and HP groups. However, the UH pups showed a tendency towards higher ACTH secretion after stress compared to H and HP groups. These results suggest that repeated pain during the first 2 weeks of life in the rat does not lead to significant changes in stress responsiveness in 2-week-old pups, but we suggest that changes in mother-pup interaction (increased grooming) might act as a buffer on the cumulative effect of pain on stress responsiveness. PMID- 12586431 TI - Transient neonatal expression of NR2B/2D subunit mRNAs of the N-methyl-D aspartate receptor in the parasympathetic preganglionic neurons in the rat spinal cord. AB - Physiological studies have shown that lower urinary tract function is regulated through glutamate receptors at the levels of spinal and supraspinal cord. Of the receptor family, N-methyl-D-aspartate (NMDA) receptors mediate activity-dependent changes of synaptic efficacy, underlying synaptic plasticity and synapse development. To know the ontogenic changes of NMDA receptor expression in the visceromotor system innervating pelvic organs, including the bladder, we employed double labeling technique of retrograde neuronal tracing and in situ hybridization for detecting NMDA subunit mRNAs in preganglionic neurons (PGNs) of the lumbosacral cord. Rats at postnatal day 7 (P7), 14 (P14), 21 (P21), and adult were used. In situ hybridization was conducted using 35S-labeled antisense oligonucleotides specific to mRNAs for NMDA receptor subunits. Hybridizing signals in PGNs were detected by a dark-field microscope equipped fluorescence detector. PGNs showed strong signals for NR1 subunit mRNA at each developmental stage examined. Moderate signals for the NR2B and NR2D subunit mRNAs were found in PGNs at P7. However, their expression levels decreased thereafter, reaching the minimal level in adults. No significant signals for NR2A and NR2C subunit mRNAs were detected at any stages. This temporal pattern of expression suggests a possible involvement of NMDA receptors in the development of micturitional neural circuit through activity-dependent mechanisms. PMID- 12586432 TI - Parallel development of blood vessels and mast cells in the lateral geniculate nuclei. AB - The present study examined quantitatively developmental changes of the vasculature in the dorsal (dLGN) and the ventral (vLGN) lateral geniculate nuclei together with concomitant changes in the number of mast cells (MCs), known for their role in angiogenesis. Vascular network, marked after transcardial perfusion of India ink, and MCs detected with conventional histochemical techniques were examined at postnatal days (P) 1, 8, 14, 21, 31, 90 and 300 of Wistar rats. Quantitative analysis by means of an image analysis system showed age-dependent changes in both vascular parameters [vascular area and relative frequency (%) of capillaries and medium- and large-diameter vessels] and mast cells number in the developing dLGN and vLGN. Despite quantitative differences in the vascularization and MC infiltration between the two nuclei at some age points, MC number, vascular area and the percentage frequency of capillaries exhibited similar developmental time courses, especially up to the end of the first postnatal month. Both MC number and the capillary frequency reached maximal levels at P31 and declined thereafter, following a massive or a partial, respectively, decrease up to P300. PMID- 12586433 TI - Neonatal focal denervation of the rat olfactory bulb alters cell structure and survival: a Golgi, Nissl and confocal study. AB - Contact between sensory axons and their targets is critical for the development and maintenance of normal neural circuits. Previous work indicates that the removal of afferent contact to the olfactory bulb affects bulb organization, neurophenotypic expression, and cell survival. The studies also suggested changes to the structure of individual cell types. The current work examines the effects of denervation on the morphology of mitral/tufted, periglomerular, and granule cells. Focal denervation drastically changed mitral/tufted cell structure but had only subtle effects on periglomerular and granule cells. Denervated mitral/tufted cells lacked apical tufts and, in most cases, a primary dendrite. In addition, the denervated cells had more secondary processes whose orientation with respect to the bulb surface was altered. Our results suggest that contact between olfactory axons and the bulb is necessary for cell maintenance and may be critical for the ability of mitral/tufted cells to achieve adult morphology PMID- 12586434 TI - Fructose-1,6-biphosphate prevents excitotoxic neuronal cell death in the neonatal mouse brain. AB - The excitotoxic cascade may represent an important pathway leading to brain damage and cerebral palsy. Brain lesions induced in newborn mice by ibotenate (acting on N-methyl-D-aspartate receptors) and by S-bromowillardiine (acting on alpha-3-amino-hydroxy-5-methyl-4-isoxazole propionic acid and kainate receptors) mimic some aspects of white matter cysts and transcortical necrosis observed in human perinatal brain damage. Fructose 1,6-biphosphate (FBP) is a high-energy glycolytic pathway intermediate which, in therapeutic doses, is non-toxic and neuroprotective in hypoxic-ischemic models of brain injury. Mechanisms of action include modulation of intracellular calcium through phospholipase C (PLC) activation. The goal of this study was to determine the neuroprotective effects of FBP in a mouse model of neonatal excitotoxic brain injury. Mice that received intraperitoneal FBP had a significant reduction in size of ibotenate-induced (80% reduction) or S-bromowillardiine-induced (40% reduction) cortical plate lesions when compared with control animals. Studies of fragmented DNA and cleaved caspase 3 confirmed the survival promoting effects of FBP. FBP had no detectable effect on excitotoxic white matter lesions. The effects of FBP were antagonized by co administration of PLC, protein kinase C or mitogen-associated protein kinase inhibitors but not by protein kinase A inhibitor. A moderate, transient cooling of pups immediately after the insult extended the therapeutic window for FBP, as FBP administered 24 h after ibotenate was still significantly neuroprotective in these pups. This data extends the neuroprotective profile of FBP in neonatal brain injury and identifies gray matter lesions involving N-methyl-D-aspartate receptors as a major target for this promising drug. PMID- 12586435 TI - A dorso-ventral gradient of Pax6 in the developing retina suggests a role in topographic map formation. AB - Expression of the transcription factor Pax6 was assessed immunohistochemically in embryonic chick retina during retino-tectal map formation. A low dorsal to high ventral gradient was found that correlated with expression of the axonal guidance cue EphB2. Furthermore, transfection of Pax6 into undifferentiated P19 cells up regulated EphB2. The results raise the possibility that Pax6 is upstream of EphB2 and that its graded expression defines the dorso-ventral axis of the retino tectal projection. PMID- 12586436 TI - Reelin is expressed in the accessory olfactory system, but is not a guidance cue for vomeronasal axons. AB - Reelin is an extracellular matrix protein that regulates neuronal migration in the developing cerebral cortex, and axon outgrowth in the hippocampus. In the developing vomeronasal system, Reelin mRNA is expressed in perineural cells near the vomeronasal nerve, as well as in the vomeronasal organ, olfactory epithelium and olfactory and accessory olfactory bulbs, suggesting that it might regulate axon guidance or fasiculation. We tested that hypothesis by crossing reeler mice with VN12-IRES-tau-lacZ mice to investigate the role of reelin. The vomeronasal nerves are indistinguishable in normal and reeler mutant mice, strongly suggesting that Reelin does not provide a guidance cue for vomeronasal axons. PMID- 12586437 TI - Postnatal development of the glutamate vesicular transporter VGLUT1 in rat cerebral cortex. AB - The expression of the vesicular glutamate transporter VGLUT1 in the rat neocortex was studied during postnatal development using immunocytochemistry and Western blotting. At all ages, VGLUT immunoreactivity is localized to puncta that coexpress the presynaptic marker synaptophysin. VGLUT1 immunoreactivity is faint at birth, increases in the subplate during the first postnatal week, invades the supragranular layers in the second week and reaches the adult pattern at P20-P30. Its spatial and temporal maturation patterns suggest that VGLUT1 may be the vesicular transporter in developing corticocortical connections. PMID- 12586438 TI - Differentiation and genetic manipulation of human embryonic stem cells and the analysis of the cardiovascular system. AB - Human embryonic stem (ES) cells are pluripotent cells derived from blastocyst stage embryos. The cells are characterized by their self-renewal capability and by their ability to differentiate into a wide range of cell types. In vivo, injection of the human ES cells into immune-deficient mice generates teratomas harboring derivatives of all three embryonic germ layers. In vitro, spontaneous aggregation of human ES cells results in the formation of embryoid bodies (EBs) comprised of differentiated cells from the three embryonic germ layers. Induced differentiation of ES cells into specific subsets of cells may be generated by treatment with several growth factors. Cardiomyocytes and endothelial cells were among the tissue types identified in vitro, and one of the most dramatic examples for the differentiation of human ES cells is the formation of rhythmic contractions of EBs containing pulsing cardiac muscle cells. Cells of the cardiovascular system were characterized by many molecular markers and by their structural and functional properties. The ability to genetically manipulate human ES cells now allows for the purification of specific cell types. Human ES cells have tremendous value as an in vitro model to study embryonic differentiation and as a source of cells for cellular transplantation in various pathologies among them cardiovascular diseases. PMID- 12586439 TI - Leucine/isoleucine zipper coordination of ion channel macromolecular signaling complexes in the heart. Roles in inherited arrhythmias. AB - The sympathetic nervous system controls the force and rate of contraction of the heart. The rapid response to stress and exercise mediated by increased sympathetic nervous system (SNS) activity requires the coordinated regulation of several ion channels in response to activation of beta-adrenergic receptors. The microenvironment of target channels is mediated by the assembly of macromolecular signaling complexes in which targeting proteins recruit phosphatases and kinases and in turn bind directly to the channel protein via highly conserved leucine/isoleucine zippers (LIZs). Disruption of local signaling by disease associated LIZ mutations unbalances the physiologic response to SNS stimulation and increases the risk of arrhythmia in mutation carriers. PMID- 12586440 TI - NFAT regulation in smooth muscle. AB - First identified in activated T cells, the calcium (Ca2+)-dependent transcription factor, nuclear factor of activated T cells (NFAT), has since been shown to play a role in nonimmune cells, including cells of the cardiovascular system. In arterial smooth muscle, the diverse array of calcium-signaling modalities, the functional interplay between smooth muscle and endothelial cells, and the influence of intravascular pressure on calcium and other signaling pathways creates a calcium-regulatory environment that is arguably unique. This review focuses on mechanisms that control the initial Ca2+/calcineurin-dependent events in NFAT activation, with a particular emphasis on NFAT regulation in native vascular smooth muscle. Also addressed is the role of additional mechanisms that act to modulate calcineurin-dependent NFAT nuclear import/export, mechanisms that may have particular relevance in this tissue. PMID- 12586441 TI - Targeting diastolic dysfunction by genetic engineering of calcium handling proteins. AB - Diastolic heart failure (HF) is associated with significant morbidity and mortality, and is a growing medical problem in this country. Diastolic dysfunction is defined as an abnormality in myocardial relaxation that impairs filling during diastole and contributes to the clinical syndrome of HF. Effective clinical strategies to treat diastolic dysfunction are limited. This article focuses on the potential application of parvalbumin--a fast skeletal muscle calcium buffer--for remediation of slow relaxation in the failing heart. PMID- 12586442 TI - A new role for corticotropin-releasing factor receptor-2: suppression of vascularization. AB - The corticotropin-releasing factor (CRF) family of receptors and ligands are known to have potent effects on vasculature function. Our recent data has described a novel role of corticotropin-releasing factor receptor-2 (CRFR2) as a tonic inhibitor of neovascularization. CRFR2 is found in both endothelial and smooth muscle cells (SMC) in the vasculature, where its function has been elusive. From investigation into the role of CRFR2 as a determinant of tissue vascularization, it was discovered that mice deficient for CRFR2 become hypervascularized postnatally and express increased levels of vascular endothelial growth factor (VEGF). Mechanistically, CRFR2 activation in vitro was found to result in reduced VEGF release from SMCs, an inhibition of SMC proliferation, and an inhibition of capillary tube formation in collagen gels. Treatment of a subcutaneously injected gel matrix with a CRFR2 agonist inhibited growth factor-induced vascularization in vivo. Western blots for the cell-cycle retinoblastoma protein (Rb)--essential for cell-cycle progression, showed decreased levels of phosphorylated Rb following CRFR2 agonist treatment in SMCs- supporting a role for CRFR2 in regulation of SMC proliferation. These results suggest that CRFR2 is a critical component of a novel pathway necessary for tonic inhibition of adult neovascularization, and that CRFR2 may be a potential target for therapeutic modulation of angiogenesis. PMID- 12586443 TI - Role of the JAK-STAT pathway in protection against myocardial ischemia/reperfusion injury. AB - The Janus kinase (JAK)-signal transducers and activators of transcription (STAT) pathway is a stress-responsive mechanism that transduces signals from the cell surface to the nucleus, thereby modulating gene expression. Recent studies have demonstrated that myocardial ischemia and reperfusion induce rapid activation of this pathway. Although the functional consequences of this event remain to be elucidated, there is emerging evidence that JAK-STAT signaling plays an important role in the development of the cardioprotected phenotype associated with ischemic preconditioning. Specifically, brief episodes of myocardial ischemia/reperfusion activate JAK1 and JAK2, followed by recruitment of STAT1 and STAT3, resulting in transcriptional upregulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), which then mediate the infarct-sparing effects of the late phase of preconditioning. The present review focuses on this novel cardioprotective role of JAK-STAT signaling and on its potential exploitation for developing therapeutic strategies aimed at limiting ischemia/reperfusion injury. PMID- 12586444 TI - ErbB2 pathways in heart and neural diseases. AB - The proto-oncogene ErbB2 (also known as c-neu or HER2 in humans) encodes a receptor tyrosine kinase that is frequently overexpressed in human tumors. It is the target of a novel and effective antibody-based therapy for malignant mammary tumors (trastuzumab/Herceptin). Biochemical and genetic experiments have shown that ErbB2 acts as a coreceptor for other members of the ErbB family of receptor tyrosine kinases. In particular, signals are transduced by ErbB2/ErbB4, ErbB2/ErbB3, and ErbB2/EGF receptor heteromers. ErbB2/4 and ErbB2/ErbB3 heteromers transmit neuregulin-1 signals in the developing and adult heart, and in the peripheral nervous system, respectively. Of particular medical relevance are recent findings that relied on tissue-specific mutation of ErbB2 in cardiomyocytes, which revealed an essential function of ErbB2 in normal heart physiology and demonstrated that loss of cardiac ErbB2 can cause dilated cardiomyopathy in adult mice. Thus, ErbB2 is important not only in development, but also for the correct functioning of the differentiated myocardium. The conditional ErbB2 mutant mice provide a model for the principal side effects- cardiomyopathy and heart failure--that can be observed in patients undergoing chemotherapy with Trastuzumab. PMID- 12586445 TI - Neuropeptide Y: a novel mechanism for ischemic angiogenesis. AB - Sympathetic nerve activation often accompanies tissue ischemia, which in turn stimulates angiogenesis, but whether the nerves regulate vascular functions beyond vasoconstriction (i.e., by promoting new vessel formation) has never been established. Neuropeptide Y (NPY) is a sympathetic cotransmitter preferentially released during intense or prolonged stress, which causes vasoconstriction and vascular smooth muscle cell proliferation by activating multiple Gi/o-coupled receptors, Y1 and Y5. At nonvasoconstrictive concentrations and through non-Y1 receptors, NPY also stimulates endothelial cell adhesion to matrix, migration, proliferation, capillary tube formation on matrigel, and aortic sprouting. Recent studies also indicate that NPY and its non-Y1 receptors exert powerful angiogenic effects in peripheral limb ischemia, promising a new way of treatment for revascularization of ischemic tissues. PMID- 12586446 TI - Induction of c-Fos expression in specific areas of the fear circuitry in rat forebrain by anxiogenic drugs. AB - BACKGROUND: The fact that induction of anxiety- and panic-related symptoms is a property common to a range of drugs suggests that common neural substrates underlie their behavioral effects. METHODS: We used Fos immunocytochemistry to test the effects of four anxiogenic drugs (FG-7142, yohimbine, m chlorophenylpiperazine [mCPP], and caffeine) on anxiety-related circuitry in rat forebrain. RESULTS: All four drugs commonly increased Fos-like immunoreactivity in 7 of 41 brain areas investigated, namely, central nucleus of the amygdala, bed nucleus of the stria terminalis, lateral septum, paraventricular nucleus of the hypothalamus, lateral hypothalamus, infralimbic and prelimbic cortex. All drugs but one (mCPP) also increased Fos expression in the basolateral and medial amygdala, the dorsomedial hypothalamus, cingulate cortex, and parts of the motor cortex. CONCLUSIONS: The results suggest that the anxiogenic drugs selected activate a restricted set of forebrain areas. Most of these areas have previously been shown to be activated by environmentally evoked anxiety and to have anatomic connections with hindbrain regions that are activated by the same drugs and by environmentally evoked anxiety. Together, these data are consistent with the theory of an integrated forebrain and hindbrain neuronal system that is important for anxiety states evoked by both drug and environmental manipulations. PMID- 12586447 TI - Heritability of fearful-anxious endophenotypes in infant rhesus macaques: a preliminary report. AB - BACKGROUND: Research efforts to discover the genetic underpinnings of anxiety and depression is challenging because of the etiologic heterogeneity inherent to these disorders. These efforts might be aided by the study of related behavioral phenotypes in model organisms, such as monkeys. METHODS: Eighty-five rhesus monkeys (Macaca mulatta) from the Oregon National Primate Research Center were drawn from a standard matriarchal colony and tested for behavioral response in four testing paradigms designed to elicit fearful-anxious reactions. Heritabilities were estimated using variance component-based quantitative genetic analyses with much of the genetic information arising from paternal half-sibs. RESULTS: Individual behaviors reflecting increased distress responses (e.g., vocalizations and teeth grinding) and behavioral inhibition (e.g., latency to leave mother, latency to inspect novel fruit) showed significant heritability, even though a small number of monkeys were assessed. Exploratory factor analyses identified seven clusters of behaviors across tests, some of which were found to be heritable. CONCLUSIONS: These results indicate that several specific fearful anxious behaviors in infant rhesus monkeys are heritable within this colony. Accordingly, these phenotypes, which are believed to represent the genetic liability for anxiety and depression, are good candidates for further genetic investigation in this population. PMID- 12586448 TI - Impact of chronic intermittent challenges in stressor-susceptible and resilient strains of mice. AB - BACKGROUND: Stressors promote altered neurochemical functioning that may be of adaptive value. The sustained elevations of neurochemical activity elicited by chronic stressors may exact excessive demands on biological systems (allostatic load), thereby rendering the organism more vulnerable to pathology, but such effects may be dependent on individual characteristics. METHODS: Stressor reactive BALB/cByJ mice and the relatively resilient C57BL/6ByJ mice were exposed to a variety of psychogenic and neurogenic stressors, twice a day over 60 days. The resultant neurochemical and behavioral changes in these strains was assessed relative to the effects of acute stressors. RESULTS: Acute restraint elicited more pronounced and more widespread variations of norepinephrine and serotonin utilization in BALB/cByJ than in C57BL/6ByJ mice. Following the chronic stressor, BALB/cByJ mice showed marked behavioral alterations thought to be indicative of depression- and anxiety-like states. The chronic stressor was also associated with moderation of amine utilization in the C57BL/6ByJ mice, whereas such an outcome was less prominent or entirely absent in BALB/cByJ mice. CONCLUSIONS: The sustained increase of amine activity in the BALB/cByJ, coupled with the marked depressionogenic-anxiogenic characteristics of this strain, may provide a useful preparation to assess the impact of chronic stressors on the development of behavioral and physical pathology. PMID- 12586449 TI - Evidence for disturbed cortical signal processing and altered serotonergic neurotransmission in generalized anxiety disorder. AB - BACKGROUND: Current pathophysiological concepts of generalized anxiety disorder (GAD) assume a disturbed exteroceptive sensory system. Furthermore, central serotonergic neurotransmission has been shown to play an important role in anxiety disorder. Cortical signal processing as measured by auditory evoked potentials (AEPs) may reflect the integrity of the exteroceptive sensory system. Because a special aspect of AEP, the loudness dependence of the N1/P2-component (LD), has been related to central serotonergic activity, the LD may be useful for investigating serotonergic dysfunctions in GAD. METHODS: The LD was recorded in 31 medication-free patients with GAD without any psychiatric co-morbidity and in 31 matched control subjects. Dipole source analysis was performed to separate the LD of regions including the primary (LD-tangential dipole) and regions including the secondary auditory cortex (LD-radial dipole). RESULTS: A shallower LD tangential was observed in patients with GAD as compared to healthy control subjects [F(1,60) = 6.727, p =.012; one-way analysis of variance]. The LD-radial showed no differences between groups. Severity of the anxiety symptoms was not related to the LDs. CONCLUSIONS: The results indicate an altered exteroceptive sensory system in GAD occurring at the level of the primary but not secondary auditory cortex. Because a shallow LD of the primary auditory cortex was related to a high firing rate of neurons in the dorsal raphe nucleus, the results may support evidence for an enhanced serotonergic activity in GAD. PMID- 12586450 TI - Elevated agonist binding to alpha2-adrenoceptors in the locus coeruleus in major depression. AB - BACKGROUND: Recent postmortem studies demonstrate disrupted neurochemistry of the noradrenergic locus coeruleus (LC) in major depression (MD). Increased levels of tyrosine hydroxylase and decreased levels of norepinephrine transporter implicate a norepinephrine deficiency in the LC in MD. Here we describe a study of alpha2 adrenoceptors in the LC and raphe nuclei of subjects with MD compared with psychiatrically normal control subjects. METHODS: The specific binding of p [125I]iodoclonidine to alpha2-adrenoceptors was measured at multiple levels along the rostrocaudal extent of the LC in postmortem tissue from 14 control and 14 MD subjects. In addition, p-[125I]iodoclonidine binding was measured in the dorsal and median raphe nuclei in the same tissue sections. RESULTS: The specific binding of p-[125I]iodoclonidine to alpha2-adrenoceptors was significantly elevated throughout the LC from MD compared with matched control subjects. No significant differences were observed in p-[125I]iodoclonidine binding to alpha2 adrenoceptors in the raphe nuclei comparing MD and control subjects. CONCLUSIONS: Given that alpha2-adrenoceptors are upregulated in laboratory animals by treatment with drugs that deplete norepinephrine, our findings implicate a premortem deficiency of brain norepinephrine in the region of the locus coeruleus in subjects with MD. PMID- 12586451 TI - A randomized controlled comparison of electroconvulsive therapy and repetitive transcranial magnetic stimulation in severe and resistant nonpsychotic major depression. AB - BACKGROUND: Studies published over the past few years suggest that transcranial magnetic stimulation (TMS) may have significant antidepressant actions. In a previous report, we compared electroconvulsive therapy (ECT) and repetitive TMS (rTMS) and found ECT to be superior for psychotic major depression (MD); however, ECT and rTMS had similar results in nonpsychotic MD. We now report on a controlled randomized comparison of ECT and rTMS in patients with nonpsychotic MD. METHODS: Forty patients with nonpsychotic MD referred for ECT were included. Electroconvulsive therapy was performed according to established protocols. Repetitive TMS was performed over the left dorsolateral prefrontal cortex at 90% motor threshold. Patients were treated with 20 sessions (five times per week for 4 weeks) of 10-Hz treatments (1200 pulses per treatment-day) at 90% motor threshold. Response to treatment was defined as a decrease of at least 50% in the Hamilton Rating Scale for Depression (HRSD) score, with a final HRSD equal or less than 10 points and a final Global Assessment of Function Scale rating of 60 or more points. RESULTS: The overall response rate was 58% (23 out of 40 patients responded to treatment). In the ECT group, 12 responded and eight did not; in the rTMS group, 11 responded and nine did not (chi2 =.10, ns). Thus, patients responded as well to either ECT or rTMS. CONCLUSIONS: This study adds to the growing literature supporting an antidepressant effect for rTMS. This study is particularly relevant because it suggests that rTMS and ECT reach similar results in nonpsychotic major depressive disorder. PMID- 12586452 TI - Effects of tryptophan depletion and catecholamine depletion on immune parameters in patients with seasonal affective disorder in remission with light therapy. AB - BACKGROUND: Altered immunologic parameters are found in symptomatic depressed patients relative to remitted depressed patients and healthy controls. We investigated whether tryptophan depletion and catecholamine depletion induce alterations in immunologic parameters in patients with seasonal affective disorder remitted on light therapy, and whether these changes are associated with changes in mood. METHODS: Remitted patients with seasonal affective disorder underwent tryptophan depletion, catecholamine depletion, and sham depletion in a prospective randomized, double-blind crossover design. Measures of depression, plasma levels of tryptophan and catecholamine metabolites, and plasma levels of cytokines (sIL-4, IL-6, neopterin, sTNF-R1 and sTNF-R2) were obtained at baseline, and 7, 24, and 30 hours after monoamine depletion. RESULTS: Tryptophan depletion decreased plasma total and free tryptophan levels; catecholamine depletion decreased plasma 3-methoxy-4-hydroxyphenylethyleneglycol and homovanillic acid levels. Tryptophan depletion and catecholamine depletion, but not sham depletion, induced a transient exacerbation of depressive symptoms (p <.001); plasma neopterin levels increased during tryptophan depletion and catecholamine depletion (p <.05). Tryptophan depletion and catecholamine depletion induced a transient reduction of plasma sIL-4 levels (p <.05). A significant correlation was found between sIL-4R levels and depression ratings after tryptophan depletion (r = -.61, p <.05). CONCLUSIONS: The monoamine depletion-induced alterations of humoral and cellular immunity suggest a potential role of immunologic parameters in the pathophysiology of seasonal affective disorder; however, the results must be considered preliminary and require further study. PMID- 12586453 TI - Larger amygdala volumes in first depressive episode as compared to recurrent major depression and healthy control subjects. AB - BACKGROUND: The aim of our study was to test the hypothesis that amygdala volumes are reduced in patients with recurrent major depression compared with first episode patients. METHODS: Using structural magnetic resonance imaging, we compared 30 inpatients with first-episode depression and 27 inpatients with recurrent major depression (DSM-IV) with healthy volunteer subjects from the local community matched for age, gender, and handedness. RESULTS: Patients with first-episode depression showed enlarged amygdala volumes compared with patients with recurrent major depression and healthy control subjects. No significant differences were found between patients with recurrent depression and healthy control subjects. No significant correlations were found between amygdala volumes and age of onset, illness duration, or severity of depression. CONCLUSIONS: Larger amygdala volumes in patients with first-episode depression may result from higher amygdala metabolism and blood flow. Additionally, disease progression with stress-related excitotoxic processes during recurrent depressive episodes might result in decreased amygdala volumes. Prospective investigations to investigate amygdala changes during the course of depression are needed. PMID- 12586454 TI - Involvement of NMDA receptors in alcohol-mediated behavior: mice with reduced affinity of the NMDA R1 glycine binding site display an attenuated sensitivity to ethanol. AB - BACKGROUND: Ethanol antagonizes central effects of glutamate by inhibition of the N-methyl-D-aspartate (NMDA) receptor function. The co-agonist glycine has been shown to reverse alcohol-mediated effects. The aim of this study was to evaluate the influence of the glycine binding site of the NMDA receptor on the behavioral effects of alcohol by investigating mice with an 80% reduced affinity of the NMDA R1 subunit for glycine (Grin1(D481N)). METHODS: Free-choice and forced alcohol intake was studied over a period of 52 days. Anxiolytic activity (elevated plus maze, open field) and motor coordination (rotarod) was tested after 3 days of forced alcohol intake and during ethanol withdrawal. RESULTS: In contrast to wild type mice, alcohol-associated anxiolysis and motor impairment was attenuated in Grin1(D481N) mice during intoxication. Free-choice alcohol intake did not differ between wild-type and Grin1(D481N) mice. CONCLUSIONS: Our results give first evidence in vivo for a possible role of an altered NMDA-receptor complex with a hyposensitive glycine binding site for behavioral effects mediated by ethanol. PMID- 12586455 TI - Valproate attenuates hyperactive and perseverative behaviors in mutant mice with a dysregulated dopamine system. AB - BACKGROUND: Dopamine transporter (DAT) knockdown (KD) mice, with approximately 90% loss of expression of the DAT, allow for the examination of the behavioral consequences of a chronically dysregulated dopamine system. The DAT KD mice have hyperdopaminergic tone, are hyperactive, and show impaired response inhibition in a number of paradigms. We hypothesized that the DAT KD mice would also display deficits in prepulse inhibition (PPI) and would be perseverative in their locomotor behavior. METHODS: Basal levels of PPI and patterns of locomotor behavior were measured in two cohorts of DAT KD mice. In addition, measurements of locomotor behavior were recorded after pretreatment with 100 mg/kg valproate in both DAT KD and wildtype mice. RESULTS: The DAT KD mice were hyperactive and displayed perseverative motor behavior but had normal levels of PPI. The clinically effective antimania drug valproate significantly attenuated the hyperactivity and perseverative locomotor behavior in the DAT KD mice and had no effect in control mice. CONCLUSIONS: The DAT KD mice appear to provide a model of some aspects of manic behavior. With limited models of bipolar disorder, the DAT KD mice might provide a vehicle to screen for new psychiatric therapies to treat mania and its related symptoms. PMID- 12586458 TI - Evaluation of matrix-assisted laser desorption ionization mass spectrometry for studying the sec-butyllithium and n-butyllithium initiated ring-opening polymerization of hexamethylcyclotrisiloxane (D3). AB - Matrix assisted laser desorption ionization (MALDI) was used to study the organolithium initiated ring-opening polymerization of hexamethylcyclotrisiloxane (D3) in a mixed solvent system. The mass spectral peak intensities were monitored to determine the effects of polymerization time, initiator concentration, and reaction temperature on the formation of the mono, di, and trisiloxanolate initiator species and the extent of chain redistribution. The three initiator species were formed by reacting n-butyllithium and sec-butyllithium with D3 in nonpolar solvent. The mass spectral results showed that sec-butyllithium and n butyllithium form different populations of initiator species under the same conditions and that the measured mass spectral peak intensities do not accurately represent the population of siloxanolate initiator species prior to propagation. The changes in peak intensities were attributed to chain redistribution. PMID- 12586459 TI - Internal energy deposition in chemical ionization/tandem mass spectrometry. AB - The efficiency of the collision-induced dissociation (CID) process as a function of the internal energy deposited into the ion during the ionization event was evaluated. (M + H)+ ions of pyrrole, pyrrolidine, pyridine and piperidine (five and six-membered ring heterocyclics) were generated by chemical ionization (CI). The internal energy of the ions was varied by using different reagent gases. Both high-energy (keV) and low-energy (eV) CID were performed on these ions. The experiments showed that the (M + H)+ ions of the five-membered ring compounds, pyrrole and pyrrolidine, have higher fragmentation efficiencies than the six membered ring compounds, pyridine and piperidine. Fragmentation efficiencies in high-energy CID clearly correlate with the internal energy deposited by the ionization technique. Experiments showed that the low-energy CID process is more sensitive than high-energy CID to changes in internal energy. PMID- 12586457 TI - H/D exchange of gas phase bradykinin ions in a linear quadrupole ion trap. AB - The gas phase H/D exchange reaction of bradykinin ions, as well as fragment ions of bradykinin generated through collisions in an orifice skimmer region, have been studied with a linear quadrupole ion trap (LIT) reflectron time-of-flight (rTOF) mass spectrometer system. The reaction in the trap takes only tens of seconds at a pressure of few mTorr of D2O or CD3OD. The exchange rate and hydrogen exchange level are not sensitive to the trapping q value over a broad range, provided q is not close to the stability boundary (q = 0.908). The relative rates and hydrogen exchange levels of protonated and sodiated +1 and +2 ions are similar to those observed previously by others with a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer system. The doubly and triply protonated ions show multimodal isotopic distributions, suggesting the presence of several different conformations. The y fragment ions show greater exchange rates and levels than a or b ions, and when water or ammonia is lost from the fragment ions, no exchange is observed. PMID- 12586460 TI - Discovery of long-lived excited electronic states of vinylchloride, vinylbromide, vinyliodide, and acrylonitrile cations. AB - Charge exchange technique has been used to detect the presence of long-lived excited electronic states of some monosubstituted ethene cations. The technique is based on the criterion that charge exchange between polyatomic species is efficient only when the energy of reaction is close to zero or negative (DeltaE < or = 0), or the exoergicity rule. The A2A' states of vinylchloride, vinylbromide, vinyliodide, and acrylonitrile cations have been found to have long lifetimes (tens of microsecond or longer) while all the excited electronic states of vinylfluoride cation have been found to undergo rapid dissociation or nonradiative relaxation to the ground state. The long-lived states found are those displaying well-resolved vibrational structures in the photoelectron spectra. In particular, these are the states generated by removal of an electron from the in-plane nonbonding p orbitals of halogens or the in-plane pi orbital of the triple bond C[triple bound]N. The present findings are very similar to those for the monosubstituted benzene cations reported previously. PMID- 12586461 TI - ESI-mass spectrometry analysis of unsubstituted and disubstituted beta cyclodextrins: fragmentation mode and identification of the AB, AC, AD regioisomers. AB - The study of unsubstituted and disubstituted beta-cyclodextrins (beta-CDs) by ESI mass spectrometry is reported, applying a cone-induced fragmentation in the presence of a twofold excess of sodium chloride, in order to gain information about the fragmentation of the different regioisomers. On the basis of the fragmentation pattern observed for the unsusbstituted beta-CD, a statistical model shows that the fragments generated by every regioisomer of a disubstituted CD (AB, AC, and AD) are expected to differ in their relative intensity and, therefore, they can be used for correctly identifying the three different regioisomers. The model was tested on the three regioisomeric (AB, AC, and AD) diamino-beta-CDs and ditosyl-beta-CD and on the AC and AD regioisomers of dimesitylenesulphonyl-beta-CD, allowing in every case through statistical analysis of the fragmentation pattern the correct assignment of every regioisomer on the basis of an ESI mass spectrum (single quadrupole analyzer, high cone voltage) of the pure compounds. The absolute intensities of the fragmentation peaks were voltage-dependent but their ratios was voltage-independent, indicating that no mass bias in peak ratios is introduced by the analyzer. Given the fast time of analysis and its general applicability, independently from the substituents, we propose our method as an easy way to identify the regioisomers of disubstituted beta-CDs. PMID- 12586462 TI - Why CH3CH3+* formation competes with H* loss from CCCO C3H6O+* isomers. AB - How formation of CH3CH3+* competes with H* loss from C3H6O+* isomers with the CCCO framework has been a puzzle of gas phase ion chemistry because the first reaction has a substantially higher threshold and a supposedly tighter transition state. These together should make CH3CH3+* formation much the slower of the two reactions at all internal energies. However, the rates of the two reactions become comparable at about 20 kJ x mol(-1) above the threshold for CH3CH3+* formation. It was recently shown that losses of atomic fragments increase in rate much more slowly with increasing internal energy than do the rates of competing dissociations to two polyatomic fragments. This occurs because fewer frequencies are substantially lowered in transition states for the former type of reaction than for the latter. The resulting lower transition state sums of states cause the rates of dissociations producing atoms as fragments to increase much more slowly than competing processes with increasing energy. Here we show that this is why CH3CH3+* formation competes with H* loss from CH3CH2CHO+*. These results further establish that the dependence on energy of the rate of a simple unimolecular dissociation is usually directly related to the number of rotational degrees of freedom in the products, a newly recognized factor in determining the dependence of unimolecular reaction rates on internal energy. PMID- 12586463 TI - Ammonium ion mediated resorcarene capsules: ESI-FTICRMS study on gas-phase structure and ammonium ion affinity of tetraethyl resorcarene and its per methylated derivative. AB - The ammonium ion binding affinities of tetraethyl resorcarene (1) and its per methylated derivative (2) were studied by electrospray ionization Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry. Ten different ammonium ions were tested as guests for the resorcarenes. A strong tendency for complex formation was observed with all ammonium ions of size and charge distribution suitable for noncovalent interactions with the cavities of the resorcarene hosts 1 and 2. Although differences in ammonium ion affinities were observed between 1 and 2 due to the dissimilar conformations, the overall tendency was that increase in the degree of substitution and the length of carbon chain of the ammonium cation facilitated the complex formation until the sterical hindrance impeded the complexation. Dimeric as well as monomeric ammonium ion complexes were formed with resorcarene 1, but resorcarene 2 was unable to form the dimeric capsules because of the lack of H-bond donor possibilities. The nature of binding of the guest was further investigated with ion-molecule reactions and by determination of the single crystal X-ray structure of host 1 complexed with tetramethyl ammonium bromide. PMID- 12586464 TI - Recognition and quantification of binary and ternary mixtures of isomeric peptides by the kinetic method: metal ion and ligand effects on the dissociation of metal-bound complexes. AB - The kinetic method is applied to differentiate and quantify mixtures of isomeric tripeptides based on the competitive dissociations of divalent metal ion-bound clusters in an ion trap mass spectrometer. This methodology is extended further to determine compositions of ternary mixtures of the isomers Gly-Gly-Ala (GGA), Ala-Gly-Gly (AGG), and Gly-Ala-Gly (GAG). This procedure also allows to perform chiral quantification of a ternary mixture of optical isomers. The divalent metal ion Ca(II) is particularly appropriate for isomeric distinction and quantification of the isobaric tripeptides Gly-Gly-Leu/Gly-Gly-Ile (GGL/GGI). Among the first-row transition metal ions, Cu(II) yields remarkably effective isomeric differentiation for both the isobaric tripeptides, GGI/GGL using GAG as the reference ligand, and the positional isomers GAG/GGA using GGI as the reference ligand. This is probably due to agostic bonding: alpha-agostic bonding occurs between Cu(II) and GAG and beta-agostic bonding between Cu(II) and GGI, each produces large but different steric effects on the stability of the Cu(II) bound dimeric clusters. These data form the basis for possible future quantitative analyses of mixtures of larger peptides such as are generated, for example, in combinatorial synthesis of peptides and peptide mimics. PMID- 12586465 TI - Gas-phase conformations of deprotonated trinucleotides (dGTT-, dTGT-, and dTTG-): the question of zwitterion formation. AB - The gas-phase conformations of a series of trinucleotides containing thymine (T) and guanine (G) bases were investigated for the possibility of zwitterion formation. Deprotonated dGTT-, dTGT-, and dTTG- ions were formed by MALDI and their collision cross-sections in helium measured by ion mobility based methods. dTGT- was theoretically modeled assuming a zwitterionic and non-zwitterionic structure while dGTT- and dTTG- were considered "control groups" and modeled only as non-zwitterions. In the zwitterion, G is protonated at the N7 site and the two neighboring phosphates are deprotonated. In the non-zwitterion, G is not protonated and only one phosphate group is deprotonated. Two conformers, whose cross-sections differ by 17 +/- 2 A2, are observed for dTGT- in the 80 K experiments. Multiple conformers are also observed for dGTT- and dTTG- at 80 K, though relative cross-section differences between the conformers could not be accurately obtained. At higher temperatures (>200 K), the conformers rapidly interconvert on the experimental time scale and a single "time-averaged" conformer is observed in the ion mobility data. Theory predicts only one low energy conformation for the zwitterionic form of dTGT- with a cross-section 8% smaller than experimental values. Additionally, the extra H+ on G does not bridge both phosphates. Thus, dTGT- does not appear to be a stable zwitterion in the gas phase. Theory does, however, predict two low-energy conformers for the non zwitterionic form of dTGT- that differ in cross-section by 18 +/- 3 A2, in good agreement with the experiment. In the smaller cross-section form (folded conformer), G and one of the T bases are stacked while the other T folds towards the stacked pair and hydrogen bonds to G. In the larger cross-section form (open conformer), the unstacked T extends away from the T/G stacked pair. Similar folded and open conformers are predicted for all three trinucleotides, regardless of which phosphate is deprotonated. PMID- 12586467 TI - Babesiosis: persistence in the face of adversity. AB - Many babesial parasites establish infections of long duration in immune hosts. Among different species, at least four mechanisms are known that could facilitate evasion of the host immune response, although no one species is (yet) known to use them all. This update strives to illustrate the ramifications of these mechanisms and the interplay between them. PMID- 12586468 TI - Characterization and role of protozoan parasite proteasomes. AB - The proteasome, a large non-lysosomal multi-subunit protease complex, is ubiquitous in eukaryotic cells. In protozoan parasites, the proteasome is involved in cell differentiation and replication, and could therefore be a promising therapeutic target. This article reviews the present knowledge of proteasomes in protozoan parasites of medical importance such as Giardia, Entamoeba, Leishmania, Trypanosoma, Plasmodium and Toxoplasma spp. PMID- 12586469 TI - Molecular monitoring in malaria vaccine trials. AB - Molecular techniques offer new approaches for malaria field trials, particularly PCR techniques, which facilitate accurate diagnosis of Plasmodium infections and increase the power of estimates of vaccine effects on malaria prevalence or incidence. Molecular methods also help to assess selective effects of vaccines. Longitudinal genotyping data can be used to initiate novel analyses of parasite dynamics, including estimates of incidence of infection with individual parasite clones and duration of infections. In addition, high-throughput methods can be used to apply these techniques routinely in randomized controlled trials, as well as programme-based evaluations of malaria control. PMID- 12586470 TI - Invasion of skin by schistosome cercariae: some neglected facts. AB - The process of skin invasion by Schistosoma cercariae was reviewed in a recent Trends Research Update, accompanied by a computer animation. Some aspects of that article were misleading and perpetuated misconceptions about parasite migration through the skin that should by now have been dispelled. This article sets out a different interpretation of events, taking account of the extensive data on migration and larval structural changes that have been documented for Schistosoma mansoni over the past 20 years. PMID- 12586472 TI - Fecal mass and parasite strategies. PMID- 12586474 TI - Malaria control and the evolution of drug resistance: an intriguing link. AB - Does antimalarial drug resistance evolve faster in areas of high or low transmission? Suggestions that resistance evolves faster in areas of low transmission cast a cloud over control measures, such as bednet provision and insecticide spraying, by implying that their impact could be offset by the enhanced evolution of drug resistance. Theoretical analyses are ambivalent on this question, but a recent field study has attempted to measure the relationship empirically, and has generated some intriguing data: antimalarial drug resistance could be inhibited in the early stages of control programmes, only starting to resurge as the disease nears eradication. PMID- 12586475 TI - Pre-erythrocytic antigens of Plasmodium falciparum: from rags to riches? AB - A growing number of Plasmodium genomes have joined the sequencing treadmill, and the genome of Plasmodium falciparum has recently been published. Most malaria vaccinologists will soon be confronted by a bewildering array of new potential antigens from the recently completed genome of this parasite. However, for those aiming to target the pre-erythrocytic stages of the hepatic parasite, the wait might be long. In the absence of readily available materials and specific reagents, the selection of pre-erythrocytic antigens from raw sequence data is likely to prove difficult. Here, current knowledge of pre-erythrocytic antigens is updated in the light of recent results, and the post-genomic prospects of completing the antigenic repertoire of these immunologically important and intriguing stages is discussed. PMID- 12586476 TI - Mobile genetic elements colonizing the genomes of metazoan parasites. AB - A substantial fraction of the genome of most eukaryotes, including those of metazoan parasites, is predicted to comprise repetitive sequences. Mobile genetic elements (MGEs) will make up much of these repetitive sequences, particularly the interspersed sequences. This article reviews information on MGEs that have colonized the genomes of metazoan parasites (i.e. parasites of parasites). Helminth and mosquito genomes, in particular, are compared with those of better understood model organisms. MGEs from the genomes of metazoan parasites can be expected to have practical uses in transgenesis and epidemiological studies. PMID- 12586477 TI - T cells as mediators of protective immunity against liver stages of Plasmodium. AB - T cells from different subsets play a major role in protective immunity against pre-erythrocytic stages of malaria parasites. Exposure of humans and animals to malaria sporozoites induces (alphabeta CD8(+) and CD4(+) T cells specific for antigens expressed in pre-erythrocytic stages of Plasmodium. These T cells inhibit parasite development in the liver, and immunization with subunit vaccines expressing the respective antigenic moieties confers protection against sporozoite challenge. gammadelta and natural killer T cells can also play a role in protective immunity. Recent studies with mice transgenic for the alphabeta T cell receptor have revealed the existence of complex mechanisms regulating the induction and development of these responses. PMID- 12586478 TI - Molecular strategies to study Plasmodium-mosquito interactions. AB - It is widely known that malaria kills millions of people every year. Less well recognized is the fact that the situation is steadily deteriorating for a lack of effective means to counter the disease. An essential first step towards the development of new approaches to fight malaria is a thorough understanding of the mechanisms that direct parasite growth and differentiation, including parasite host interactions. This article reviews recent achievements and introduces some promising new technologies and approaches for studying host-parasite interactions. PMID- 12586479 TI - IgG subclass responses in a longitudinal study of canine visceral leishmaniasis. AB - Visceral leishmaniasis (VL), caused by Leishmania infantum, is an important disease of domestic dogs. Here, we present data on the IgG subclass antibody response to crude L. infantum antigen in a cohort of naturally infected Brazilian dogs. Specific IgG1-IgG4 responses could be detected in 98, 58, 70 and 82%, respectively of 57 dogs that were seropositive for specific IgG. Levels of all IgG subclasses were strongly inter-correlated. Levels of all IgG subclasses increased at the time of seroconversion, before reaching a plateau after several months. Levels of all IgG subclasses were higher in sick dogs than healthy dogs, and levels of all except IgG2 were higher in parasite-positive (by PCR) than parasite-negative dogs. However, levels of IgG2 relative to IgG1 were lower in sick or parasite-positive dogs compared to healthy or parasite-negative infected dogs. In contrast to previous studies, the results suggest that canine VL is associated with upregulation of specific antibody of all IgG subclasses, particularly IgG1, IgG3 and IgG4. PMID- 12586480 TI - Immunopathological studies on feline cutaneous and (muco)cutaneous mycobacteriosis. AB - Eight cases of feline (muco)cutaneous mycobacteriosis were studied to identify the causative agent and examine for phenotype and functional characteristics (expression of interleukin (IL)-1beta, IL-6, IL-12, tumour necrosis factor-alpha and inducible nitric oxide synthase) of the inflammatory cells. Polymerase chain reaction and sequencing identified the causative agents as Mycobacterium tuberculosis or M. avium complex in each four cases. Lesions were characterised by pyogranulomatous infiltration, with variability in the presence and size of necrotic areas, the presence of multinucleated giant cells and the degree of lymphocyte infiltration. Macrophages were positive for myeloid/histiocyte antigen (calprotectin), suggesting they represented freshly recruited monocytes; further differentiation to epithelioid cells and multinucleated giant cells was associated with loss of the myeloid/histiocyte antigen. Lymphocytes were found disseminated in the infiltrate (predominantly T cells) and as B cell-dominated accumulations mainly in the periphery of the lesions. Acid-fast bacilli were numerous. In M. tuberculosis complex infection, extracellular bacilli were most prominent, whereas in M. avium complex infection, bacilli were mainly located intracellularly. All cytokines examined as well as inducible nitric oxide synthase (iNOS) were variably expressed by macrophages, epithelioid cells and multinucleated giant cells. Expression was most intense in degenerating macrophages loaded with intracellular bacilli, but was also seen cell-free within necrotic areas. The intense induction of cytokine and iNOS expression especially in infected macrophages suggests a relatively low virulence for these infectious agents in cats. Furthermore, the confinement of the bacilli to lesions indicates a successful response to infection. PMID- 12586481 TI - Characterisation of porcine monocyte-derived dendritic cells according to their cytokine profile. AB - The influence of interferon (IFN)-alpha on the in vitro differentiation of myeloid porcine dendritic cells (DC) was evaluated as the ability of the DC to stimulate to cell proliferation in a mixed leukocyte reaction (MLR), and as their ability to produce cytokines at exposure to bacterial and viral preparations. Porcine monocytes were enriched from purified peripheral blood mononuclear cells (PBMC) by plastic adherence and cultured in the presence of granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 or in GM CSF, IL-4 and IFN-alpha. After 5 days of culture, the cells developed a dendritic morphology and the proportion of cells expressing MHC class II and B7 molecules was increased as determined by flow cytometry. Dendritic cells, differentiated for 5 days in GM-CSF, IL-4 and IFN-alpha, were able to stimulate both allogeneic and syngeneic PBMC to proliferation in an MLR. The DC produced the Th1 associated cytokines IFN-alpha at Sendai virus stimulation, and IL-12 at stimulation with plasmid DNA (pre-incubated in the presence of lipofectin), heat-inactivated Actinobacillus pleuropneumoniae, UV-inactivated Aujeszky's disease virus and live Sendai virus. The heat-inactivated bacteria and Sendai virus also induced production of the Th2 associated cytokines IL-10 and IL-6. The addition of IFN alpha during differentiation of DC in GM-CSF and IL-4 enhanced their ability to stimulate allogeneic and syngeneic MLR, but did not alter their ability to produce cytokines. PMID- 12586482 TI - Synovial MMP-3 and TIMP-1 levels and their correlation with cytokine expression in canine rheumatoid arthritis. AB - The purpose of this study was to investigate whether synovial fluid levels of matrix metalloprotease-3 (MMP-3) and tissue inhibitor of metalloprotease-1 (TIMP 1) are specifically elevated in canine rheumatoid arthritis (CRA) compared to osteoarthritic joint disorders and if these markers are correlated with a specific pattern of cytokine mRNA expression. Synovial fluid samples of 17 dogs with CRA were analysed for MMP-3 and TIMP-1 by two canine sandwich ELISA (enzyme linked immunosorbent assay) systems. The synovial mRNA content of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin 12 (IL-12), transforming growth factor-beta (TGF-beta), and interferon-gamma (IFN gamma) was determined by RT-PCR (reverse transcription-polymerase chain reaction). Dogs with osteoarthritis (n = 50) caused by anterior cruciate ligament rupture (ACLR) were used as controls. A significant rise of MMP-3 was found in the synovial fluid of joints with CRA that could not be balanced by sufficient amounts of TIMP-1. The 30-fold surplus of MMP-3 over TIMP-1 was strongly correlated with the synovial mRNA content of IL-1, IL-12 and TGF-beta. Our results point to the potential use of the synovial levels of MMP-3 as a marker for RA in dogs. PMID- 12586483 TI - Differentiation of porcine dendritic cells by granulocyte-macrophage colony stimulating factor expressed in Pichia pastoris. AB - Dendritic cells (DC) are potent inducers of acquired immunity due to their ability to present antigens in the context of a costimulatory environment and consequently serve an essential role in vaccine efficacy. Strategies to enhance their function, such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4 treatment to induce DC differentiation from peripheral blood monocytes, may therefore be useful as vaccine adjuvants. We now have evaluated the effect of recombinant GM-CSF on the differentiation of DC in swine. GM-CSF mRNA was readily detected in porcine splenocytes, with increased levels following treatment of the cells with ConA and LPS. Porcine GM-CSF was cloned and expressed in the methylotrophic yeast, Pichia pastoris, as a glycosylated protein that induced proliferation of porcine bone marrow cells. P. pastoris-derived GM-CSF induced expression of antigen presenting (MHC class II) and costimulatory (CD80-CD86) molecules and enhanced antigen presenting cell (APC) function consistent with the induction of functional DC. Thus, recombinant GM-CSF produced by P. pastoris may be a potent adjuvant for swine vaccines. PMID- 12586484 TI - Development of an immunoassay to measure the humoral immune response of hybrid striped bass Morone chrysops x M. saxatilis. AB - Hybrid striped bass (HSB) were immunized with bovine serum albumin (BSA) and the specific anti-BSA immunoglobulin (Ig) was affinity purified from the resulting serum by means of an agarose gel-BSA column. The native Ig had an apparent molecular size of 893 KDD, by size exclusion chromatography, and when examined by polyacrylamide gel electrophoresis (PAGE) under denaturing conditions, resolved to heavy (H) and light (L) chains of 76 and 27 KDD, respectively. Affinity purified native HSB Ig was used to immunize a goat which produced specific anti HSB Ig antibody (Ab). Purified native HSB Ig was also used to produce two murine monoclonal antibodies (mAbs) with specific affinities for H and L chain moieties of the HSB Ig molecule. Both polyclonal and monoclonal antibodies could be used individually in an indirect enzyme-linked immunosorbent assay (ELISA) to measure specific anti-BSA Ig in HSB serum. These antibodies could also be used in combination to measure total Ig in a capture ELISA format. Using both assays, the kinetics of the humoral immune response of HSB was measured for 98 days following two injections of BSA. PMID- 12586485 TI - Sequence and expression of the FcRn in the porcine mammary gland. AB - Transport of immunoglobulin G across epithelial cell barriers is thought to occur by a system involving the Fcgamma receptor called the neonatal Fc receptor (FcRn). The FcRn may also play a role in IgG transport in the mammary gland. To determine the presence of FcRn in the porcine mammary gland, biopsies were taken from glands 3 days prepartum and on the day of farrowing. The full length porcine FcRn cDNA sequence was obtained by rapid amplification of cDNA ends and determined to be 1557 base pairs in length that codes for a 359 amino acid peptide. Expression of FcRn mRNA in the porcine mammary gland was determined by reverse transcriptase-PCR and revealed that the mRNA is present prepartum and on the day of farrowing. These results indicate that the FcRn is expressed in porcine mammary tissue and are consistent with the hypothesis that FcRn may have a role in mammary gland immunoglobulin transport during colostrogenesis. PMID- 12586486 TI - Advances in equine immunology: Havemeyer workshop reports from Santa Fe, New Mexico, and Hortobagy, Hungary. AB - The horse has been human kind's most important partner throughout history. Similarly, in the field of immunology, many critical scientific advances have depended on the horse. Equine immunology today is an active and important field of study, with a focus on control of many common infectious diseases and immunopathologic conditions of broad comparative interest. In 2001 two major equine immunology workshops were held, in Santa Fe, USA, and in Hortobagy, Hungary, with major sponsorship from the Havemeyer Foundation. This report summarizes the scientific themes and foci of those meetings. PMID- 12586487 TI - Reproductive performance in Ohio dairy herds in the 1990s. AB - The objective of this study was to evaluate trends in reproductive performance in Ohio dairy herds during the 1990s. Stringent criteria for herd inclusion were applied to DHI records to ensure high quality and reliability of the data. The final data set contained 44,425 monthly herd summary records from 1772 herds from years 1992, 1994, 1996 and 1998. Outcomes of interest were calving interval length, calving-to-conception interval, days to first breeding, services per conception and first service conception rate. PROC MIXED in SAS (version 8.1) was used and the monthly records were treated as repeated measures nested within years and herds. First-order autoregressive covariance structure was used to model the covariance between the repeated measures within herds. The data were analyzed for the entire herd as well as for the first lactation cows separately. During the 1990s reproductive performance declined in Ohio dairy herds. Herd average calving interval lengthened from 13.6 to 14.1 months and calving-to conception interval increased from 136 to 150 days between 1992 and 1998, respectively. Days to first service and number of services increased while first service conception rate declined on a herd level. The reproductive performance of first lactation cows, however, remained constant during this same period. Overall herd reproductive performance was associated with herd size and fat-corrected rolling herd average milk production (FCM). As milk yield increased, first service conception rate decreased and services per conception increased. Larger herds had lower first service conception rate and needed more services per conception, but they started breeding their cows earlier resulting in a slightly shorter calving interval and calving-to-conception interval compared to smaller herds. PMID- 12586488 TI - Relationship between endotoxin and prostaglandin (PGE2 and PGFM) concentrations and ovarian function in dairy cows with puerperal endometritis. AB - Blood concentrations of progesterone, 13,14-dihydro-15-keto-prostaglandin F2alpha (PGFM) and endotoxin, and uterine fluid concentrations of prostaglandin E(2) (PGE(2)), PGFM and endotoxin were evaluated in 14 dairy cows with puerperal endometritis (mild (n=6) and heavy (n=8)). Endotoxin was measured using a quantitative kinetic assay. Cows with heavy endometritis had significantly higher concentrations of plasma PGFM (P<0.01) and uterine fluid PGE(2) and endotoxin (P<0.05) than cows with mild endometritis. Concentrations of PGFM in plasma and uterine fluid, of PGFM and PGE(2), and PGE(2) and endotoxin in uterine fluid were positively and significantly (P<0.05) correlated. The presence of endotoxin in plasma was detected in one out of six mild and in eight out of eight heavy endometritis cows. Peak plasma endotoxin concentrations (0.08-9.14 endotoxin units/ml (EU/ml) were observed between 1 and 12 days postpartum (pp) and thereafter amounts generally remained below 0.1 EU/ml (last day of detection: Day 27 pp). Abnormal ovarian function was observed in six cows (four with prolonged anoestrus and two with long luteal phase after the first postpartum ovulation). Plasma endotoxin concentrations were detected in the anoestric cows. The results suggest that: (i) concentrations of uterine fluid endotoxin and PGE(2) and of plasma PGFM are related to the degree of endometritis; (ii) absorption of endotoxin from the uterus to the bloodstream occurs, mainly in heavy endometritis cows; and (iii) there is a relationship between uterine infection, endotoxin production and resumption of pp ovarian activity. PMID- 12586489 TI - Collection of oocytes through transvaginal ultrasound-guided aspiration of follicles in an Indian breed of cattle. AB - The present study was undertaken in Karan Fries, an Indian breed of cattle to (1) determine the number of follicles available for puncture and (2) explore the potential of this breed as a donor of developmentally competent oocytes. Ovum pick-up (OPU) was performed using an ultrasound machine with a transvaginal convex transducer (5 MHz) with a needle guide, single lumen 19-gauge 60 cm long needle and a vacuum pressure of 90 mmHg. The number and size of follicles in each ovary was determined before puncture. The follicles were characterized on the basis of their diameter as small (3-5 mm), medium (6-9 mm) and large (>/=10 mm). The oocytes recovered were classified by quality. They were matured in vitro, irrespective of their grade, in 50 microl droplets of the in vitro maturation (IVM) medium (TCM-199+10% fetal bovine serum(FBS)+5 microg/ml follicle stimulating hormone (folltropin)+1 microg/ml estradiol-17beta+0.2 mM sodium pyruvate), covered with paraffin oil, in 35 mm petridish for 24 h in a CO(2) incubator (5% CO(2) in air) at 38.5 degrees C. The cleavage rate was recorded at day 2 post-insemination after subjecting the oocytes to in vitro fertilization (IVF). The differences in follicular populations of all size categories among individual donors were not significant. A total of 92 oocytes were recovered by aspiration of 157 follicles, with an overall recovery rate of 59% (range 35-79%). Of these, 32% were of grades A and B and the rest of grades C and D. The mean numbers of total follicles and the oocytes recovered per session did not differ significantly among individual donors. Out of the 73 oocytes subjected to IVM and IVF, 24 reached 2-4 cell stage at day 2 post-fertilization, with a cleavage rate of 33%. The total number of oocytes recovered was correlated with the number of small (R=0.54, P<0.01) but not with the number of medium and large follicles. This study demonstrates the use of OPU as a means of obtaining developmentally competent oocytes from an Indian breed of cattle for obtaining cattle oocytes in India where cow slaughter is not allowed for religious reasons. PMID- 12586490 TI - Synchronisation of oestrus in dairy cows using prostaglandin F2alpha, gonadotrophin-releasing hormone, and oestradiol cypionate. AB - An oestrous synchronisation protocol was developed for use in lactating dairy cows using PGF(2alpha), GnRH, and oestradiol cypionate (ECP). In experiment 1, lactating dairy cows received two injections of PGF(2alpha) (on days 0 and 11) (PP; n=10) or two injections of PGF(2alpha) (days 0 and 11) and 100 microg of GnRH on day 3 (PGP; n=10). In experiment 2, cows were treated with PGP (n=7), or PGP and 1 mg of ECP at the same time (PGPE(0); n=7) or 1 day after the second PGF(2alpha) injection (PGPE(1); n=7). In experiment 3, 101 lactating dairy cows in a commercial herd were assigned to one of three treatments; PP, PGP, or PGPE(1). Follicular growth was measured by ultrasound in experiments 1 and 2. Every cow (experiments 1, 2, and 3) was blood sampled at selected intervals for progesterone and oestradiol assays and inseminated at oestrus. In experiment 1, a higher percentage of GnRH-treated cows ovulated after the first PGF(2alpha) injection (90% versus 50%; P<0.05). The GnRH-treated cows tended to have a larger dominant follicle present at the time of the second PGF(2alpha) injection (16.5+/ 0.5 mm versus 15.0+/-0.7 mm; P<0.10). The percentage of cows that ovulated after the second PGF(2alpha) injection was similar (60%). In experiment 2, cows treated with ECP had higher peak preovulatory concentrations of oestradiol in plasma (6.99+/-0.63 versus 3.63+/-0.63; P<0.01) following the second PGF(2alpha) injection and a higher percentage ovulated (86% versus 43%; P<0.05). A higher percentage of PGPE(1)-treated cows in experiment 3 were observed in standing oestrus and ovulated after the second PGF(2alpha) injection (standing oestrus, 26.4, 34.3, and 62.6%, P<0.01; ovulated, 56, 63, and 78%, P<0.05; PP, PGP, and PGPE(1), respectively). In conclusion, the PGP protocol increased the number of cows that ovulated after the first PGF(2alpha) injection and produced a more mature dominant follicle at the time of the second PGF(2alpha) injection. Adding ECP to PGP (PGPE(1)) enhanced the expression of oestrus and increased ovulation percentage. The combination of PGP and ECP is potentially a new method to routinely synchronise oestrus and ovulation in dairy cows. PMID- 12586491 TI - Effect of treatment with progesterone and oestradiol when starting treatment with an intravaginal progesterone releasing insert on ovarian follicular development and hormonal concentrations in Holstein cows. AB - Ovarian follicular development and concentrations of gonadotrophin and steroid hormones were studied in non-lactating Holstein cows following administration of progesterone (P(4)) or oestradiol benzoate (ODB) at the start of treatment with an intravaginal progesterone releasing insert (IVP(4)) in a 2 by 2 factorial experiment. Cows were treated at random stages of the oestrous cycle with an IVP(4) device (Day 0) and either no other treatment (n=8), 200 mg of P(4) IM (n=9), 2.0 mg of ODB IM (n=8) or both P(4) and ODB (n=9). Seven days later devices were removed and PGF(2alpha) was administered. Twenty-four hours later 1.0mg of ODB was administered IM. Oestrus was detected in 97.1% and ovulation in 64.7% (effect of treatment, P=0.41) of cows within 96 h of removing inserts. In the cows that ovulated, day of emergence of the ovulatory follicle was delayed (P<0.01) and more precise (P<0.05) in cows treated with ODB compared to the cows treated with P(4). Interval from wave emergence to ovulation and the diameter of the ovulatory follicle was less (P<0.05) in cows treated with ODB compared to cows treated with P(4). Combined treatment with P(4) and ODB at the time of starting treatment with an IVP(4) device did not significantly change the pattern of ovarian follicular development compared to treatment with ODB alone. Concentrations of LH and FSH in plasma were less in cows treated with ODB between Days 0 and 4 (P<0.05) while treatment with P(4) increased concentrations of FSH in plasma between Days 0 and 4 (P<0.05). When anovulatory cows were compared to ovulatory cows, diameters of follicles (P<0.001) and growth rate of follicles (P<0.01) were less in anovulatory cows between Days 7 and 9, while concentrations of FSH in plasma were greater (P<0.01), concentrations of LH similar (P>0.90) and concentrations of oestradiol were less (P=0.01) in the anovulatory cows between Days 4 and 10. Our findings support a hypothesis that ovarian follicular development following administration of P(4) or ODB at the start of treatment with an IVP(4) device differs. Anovulatory oestrus may have been associated with reduced maturity and/or later emergence of ovarian follicles. PMID- 12586492 TI - Effects of acetylsalicylic acid and metamizol on hyaluronidase activity and sperm characteristics in rams. AB - The effects of acetylsalicylic acid and metamizol on hyaluronidase activity of semen and sperm characteristics in rams were investigated. Acetylsalicylic acid and metamizol at the doses of 75 and 50 mg/kg were administered to the rams, respectively and then semen samples were taken at 1, 2, 4, 24, 48, 96, 120 and 144 h. The hyaluronidase activities of semen in rams treated with acetylsalicylic acid and metamizol were determined to increase significantly (P<0.001) when compared with control groups at all times. Additionally, the spermatozoa motilities in both groups were measured to increase significantly (P<0.05) when compared with control group. Furthermore, there were significant (P<0.01, <0.05) decreases in the sperm concentrations and semen volumes of rams treated with acetylsalicylic acid and metamizol at all times, respectively. In conclusion, although the use of acetylsalicylic acid and metamizol cause an increase in the hyaluronidase activities and spermatozoa motilities, these drugs decrease the sperm concentrations and semen volumes along 6 days. For these reason, the use of these drugs in breeding rams during ramming season is not suitable. PMID- 12586493 TI - Evaluation of cryopreserved stallion semen from Tori and Estonian breeds using CASA and flow cytometry. AB - Methods to evaluate the quality of frozen-thawed stallion semen are still needed, particularly those considering the sperm function. The present study evaluated sperm motility, membrane and acrosome integrity and the capacitation status of frozen-thawed spermatozoa from seven Tori and six Estonian breed stallions by way of computer assisted sperm analysis (CASA), a triple fluorophore stain combination and Merocyanine 540, respectively, the latter ones using flow cytometry. Two ejaculates from each stallion were cryopreserved using the Hannover method in 0.5 ml plastic straws. Two straws per ejaculate per stallion were thawed at 37 degrees C for 30s. Motility was analysed with CASA immediately after thawing, while for flow cytometry spermatozoa were cleansed by 70:40% Percoll discontinuous density gradient separation before analysed for sperm viability, acrosome integrity (stained with SNARF, PI and FITC-PSA) and capacitation status (stained with Merocyanine 540/Yo-Pro-1). Results (as least square means) were as follows: the motility of frozen-thawed semen was 43.4% for Tori stallions and 42.3% for Estonian stallions (P>0.05). After Percoll separation 79.3% of the spermatozoa from Tori stallions had intact acrosomes and 1.7% of them showed early signs of capacitation. The same parameters for Estonian stallions were 84.5 and 2.3%, respectively. There were no statistically significant differences between breeds or ejaculates within breed for any evaluated parameter. We conclude that triple staining and flow cytometry are valuable techniques to evaluate frozen-thawed stallion spermatozoa, and that no differences in quality of frozen semen were registered between Tori and Estonian breed stallions, allowing implementation of this technology in the Estonian horse population. PMID- 12586494 TI - Effect of daily clenbuterol and exogenous melatonin treatment on body fat, serum leptin and the expression of seasonal anestrus in the mare. AB - A small percentage of mature mares continue to exhibit estrous cyclicity during the non-breeding season which is of interest because of the importance of timing of the breeding season to the equine breeding industry. Previously, it was demonstrated that the continuation of estrous cycles was more likely to occur in mature than young mares. Additionally, an apparent association exists between elevated body fat and increased circulating concentrations of leptin, and the occurrence of estrous cycles during the non-breeding season. Two experiments were conducted to test the hypothesis that pharmacological manipulation of body fat and leptin by administration of the beta(2)-adrenergic receptor agonist clenbuterol, with and without continuous treatment with melatonin, would increase the proportion of mares entering anestrus or advance its timing. In Experiment 1, eight mature mares were administered clenbuterol (3.2 microg/kg) daily from October through January and six mares were untreated. In Experiment 2, eight mares were administered clenbuterol daily from June through December, eight mares were treated with continuous release melatonin implants, and seven mares were treated with both clenbuterol and melatonin. Melatonin treatment was included to confirm previous results that constant treatment with melatonin did not advance the timing of anestrus. In both studies, leptin concentrations (P<0.01, both experiments) and body fat percentage (P<0.01, Experiment 1; P<0.05, Experiment 2) declined in response to daily clenbuterol treatment. Unexpectedly, however, this decrease was attenuated in mares also treated with melatonin (P<0.05). Although treatment with clenbuterol was associated with a decline in body fat and circulating concentrations of leptin, the timing or proportion of mares exhibiting anestrus was not modified (P>0.05). The results demonstrate the manipulation of body condition via pharmacological intervention does not modify the mechanisms controlling seasonal anestrus in the mare. PMID- 12586495 TI - Influence of pre-ovulatory insemination and early pregnancy on the distribution of CD2, CD4, CD8 and MHC class II expressing cells in the sow endometrium. AB - The aim was to investigate the distribution of CD2(+), CD4(+) and CD8(+) lymphocyte subpopulations and MHC class II expressing cells in the sow endometrium following pre-ovulatory insemination and during early pregnancy. Crossbred multiparous sows (Swedish Landrace x Swedish Yorkshire) were inseminated once at 15-20 h before ovulation. The sows were slaughtered at 5-6h (group I, n=4) after AI or at 20-25 h (group II, n=4) and 70 h (group III, n=4) after ovulation, day 11 (group IV, day 1=first day of standing oestrus, n=3) and day 19 (group V, n=3). Uterine horns were flushed to control for the presence of spermatozoa and neutrophils (groups I-IV) and/or for recovery of oocytes and/or embryos (groups II-IV, control of pregnancy). Cryofixed mesometrial uterine samples were analysed by immunohistochemistry with an avidin-biotin-peroxidase method using monoclonal antibodies to lymphocyte subpopulations and MHC class II molecules. The surface (SE) and glandular (GE) epithelia as well as connective tissue layers in subepithelial (SL) and glandular (GL) areas were examined by light microscopy. Taking all groups and different tissue layers together, the most commonly observed positive cells were CD2(+) cells (P500 ng ml(-1)) over 9 h. In comparison to non-gastroretentive CR-particles and oral solution, mean absorption time was significantly extended. These outcomes demonstrate that the CR-GRDF may be used to improve levodopa therapy and can be applied to extend the absorption of other narrow absorption window drugs that require continuous input. PMID- 12586510 TI - Hydrophilic and hydrophobic cyclodextrins in a new sustained release oral formulation of nicardipine: in vitro evaluation and bioavailability studies in rabbits. AB - The feasibility of using complexes with cyclodextrins (CDs) in nicardipine (NC) controlled delivery has been examined, with a view to extending the pharmaceutical applications spectrum of these carriers. For a fast release fraction, a hydrophilic beta-cyclodextrin derivative (hydroxypropyl-beta cyclodextrin) was employed to form a water-soluble complex. For the sustained releasing portion, triacetyl-beta-cyclodextrin (TAbetaCD) was used to provide complexes with appropriate hydrophobicity. An optimal formulation was designed by the combination of each fraction in different mixing ratios. The release behaviour of the complexes, as well as of their mixtures, was examined in simulated gastric (pH 1.2) and intestinal (pH 6.8) fluids. The formulations released the drug rapidly at the initial stage, followed by a slow release. The drug release rate was markedly retarded in the increasing order of the amount of NC/TAbetaCD complex. When NC was administered to rabbits, its absorption was very rapid with a short elimination half-life, while a prolonged maintenance of the plasma levels was obtained for the two selected formulations. The drug bioavailability was considerably improved especially after the administration of the mixture of hydrophilic and hydrophobic complexes, when compared with the NC/TAbetaCD complex. The results suggested that the critical combination of hydrophilic and hydrophobic CDs complexes, in appropriate ratios, could be a promising drug delivery system with a prolonged therapeutic effect coupled with a more balanced bioavailability. PMID- 12586511 TI - Poly(ethylene glycol) as stabilizer and emulsifying agent: a novel stabilization approach preventing aggregation and inactivation of proteins upon encapsulation in bioerodible polyester microspheres. AB - Protein aggregation and inactivation are major problems associated with the encapsulation of pharmaceutical proteins in biodegradable microspheres. The objectives of this study were to identify the causes of aggregation and inactivation of two model enzymes upon solid-in-oil-in-water (s/o/w) encapsulation in poly(lactic-co-glycolic) acid (PLGA) microspheres in order to rationally develop approaches assuring their stability. S/o/w encapsulation of gamma-chymotrypsin in PLGA microspheres caused aggregation of ca. 30% and halved its specific activity. Co-lyophilization with poly(ethylene glycol) (PEG) substantially reduced the loss in enzyme activity but 8% of the protein still aggregated during encapsulation. Model studies performed under conditions relevant to the encapsulation procedure allowed pinpointing the cause of gamma chymotrypsin instability, which was mainly the formation of the oil-in-water emulsion. To prevent aggregation in this encapsulation step, the most commonly used emulsifying agent polyvinyl alcohol (PVA) was replaced by PEG because it is known to reduce protein aggregation at interfaces. The use of PEG as the emulsifying agent in the aqueous and organic phase prevented gamma-chymotrypsin inactivation and aggregation during encapsulation. The stabilization approach also worked for the model protein horseradish peroxidase and thus is of a general nature. PMID- 12586512 TI - Development and validation of an in vitro-in vivo correlation for buspirone hydrochloride extended release tablets. AB - The aim of this study was to develop an in-vitro-in-vivo correlation (IVIVC) for two buspirone hydrochloride extended release formulations and to compare their plasma concentrations over time with the commercially available immediate release (IR) tablets. In vitro release rate data were obtained for each formulation using the USP Apparatus 2, paddle stirrer at 50 and 100 rpm in 0.1 M HCl and pH 6.8 phosphate buffer. A three-way crossover study in 18 healthy subjects studied a 30 mg "Fast" (12 h) and 30 mg "Slow" (24 h) formulation of buspirone hydrochloride given once a day, and 2x15 mg immediate release tablets dosed at a 12 h interval. The similarity factor (f(2)) was used to analyze the dissolution data. A linear correlation model was developed using percent absorbed data and percent dissolved data from the two formulations. Predicted buspirone hydrochloride concentrations were obtained by use of a curve fitting equation for the immediate release data to determine the volume of distribution and fraction absorbed constants. Prediction errors were estimated for C(max) and area under the curve (AUC) to determine the validity of the correlation. pH 6.8 at 50 rpm was found to be the most discriminating dissolution method. Linear regression analyses of the mean percentage of dose absorbed versus the mean in vitro release resulted in a significant correlation (r(2)>0.95) for the two formulations. An average percent prediction error for C(max) was -0.16%, but was 16.1%, for the AUCs of the two formulations. PMID- 12586513 TI - Galactose-PEI-DNA complexes for targeted gene delivery: degree of substitution affects complex size and transfection efficiency. AB - Complexes of galactosylated polyethylenimines (gal-PEI) with DNA have been proposed for gene delivery to hepatocytes. We synthesized gal-PEI with a broad range of degrees of substitution (DS) ranging from 3.5 to 31% of all PEI amino groups by reductive amination to determine physico-chemical and biological properties with respect to the DS. Gel retardation assay for herring testes DNA polymer polyplexes showed that increasing DS compromised DNA complexation and especially condensation. Using photon correlation spectroscopy, gal-PEI complexes formed with plasmid DNA were found to increase in size with increasing galactosylation (156+/-7 nm for 0%, 486+/-76 nm for 3.5%, 467+/-86 nm for 9.7% and 652+/-123 nm for 31% DS). Zeta potentials decreased in inverse proportion to DS (0%: 30+/-3 mV, 3.5%: 22+/-2 mV, 9.7%: 15+/-1 mV, 31%: -26+/-3.5 mV) suggesting a shielding effect by carbohydrate coupling. Cytotoxicity of gal-PEI was found to decrease with increasing galactosylation (MTT and LDH assay), no toxicity was detectable for polyplexes with plasmid DNA (LDH assay). The transfection efficiency of a reporter gene complexed with gal-PEI in a hepatocyte cell culture model (HepG2) expressing the asialoglycoprotein receptor was slightly but not significantly increased for galactosylated PEIs at a nitrogen to phosphate (N/P) ratio of 2 and strongly reduced at higher N/P ratios, compatible with only a minor targeting efficiency, strongly affected by DS. In NIH-3T3 mouse fibroblasts, increasing the DS led to a decreased transfection efficiency for all N/P ratios. Our study highlights the necessity of careful optimization of polyplex composition for active gene targeting. PMID- 12586514 TI - Different synergistic roles of small polyethylenimine and Dosper in gene delivery. AB - Low-molecular-weight PEIs and cationic liposomes can be combined resulting in a synergistic increase in transfection efficiency as we have reported earlier. Here, we have further investigated the potential mechanisms of this synergy. Complex morphology, complex sizes and DNA condensation were studied using transmission electron microscopy, light scattering methods and ethidium bromide exclusion, respectively. Cellular uptake, transfection efficiency, and effect of proton pump inhibitor bafilomycin A1 were examined in cell cultures. The cellular uptake of DNA was negligible with PEI2K-DNA complexes, whereas the uptake of the PEI2K-DNA-Dosper or the Dosper-DNA complexes was maximally about 40%. The number of transfected cells was two times higher with PEI2K-DNA-Dosper complexes than with Dosper-DNA complexes. The PEI2K-DNA-Dosper combination was slightly less sensitive to bafilomycin A1 than the PEI25K-DNA or Dosper-DNA complexes. There were no differences between PEI2K and PEI25K in DNA condensation. Dosper condensed DNA slightly more in PEI2K complexes. The PEI25K-DNA complexes were much smaller (<250 nm) than the PEI2K-DNA complexes (0.5-12 micro m) which were also rather polydisperse. It is suggested that two independent mechanisms would lead to synergistic transfection efficiency: (1) Dosper improves the cellular uptake of PEI2K-DNA complexes, and (2) PEI2K improves a transfer of the complexes from lysosomes to nucleus. PMID- 12586516 TI - Reliability of burst superimposed technique to assess central activation failure during fatiguing contraction. AB - Recording a superimposed electrically-induced contraction at the limit of endurance during voluntary contraction is used as an indicator of failure of muscle activation by the central nervous system and discards the existence of peripheral muscle fatigue. We questioned on the reliability of this method by using other means to explore peripheral muscle failure. Fifteen normal subjects sustained handgrip at 60% of maximal voluntary contraction (MVC) until exhaustion. During sustained contraction, the power spectrum analysis of the flexor digitorum surface electromyogram allowed us to calculate the leftward shift of median frequency (MF). A superimposed 60 Hz 3 s pulse train (burst superimposition) was delivered to the muscle when force levelled off close to the preset value. Immediately after the fatigue trial had ended, the subject was asked to perform a 5 s 60% MVC and we measured the peak contractile response to a 60 Hz 3 s burst stimulation. Recordings of the compound evoked muscle action potential (M-wave) allowed us to explore an impairment of neuromuscular propagation. A superimposed contraction was measured in 7 subjects in their two forearms, whereas it was absent in the 8 others. Despite these discrepancies, all subjects were able to reproduce a 3 s 60% MVC immediately after the fatigue trial ended and there was no post-fatigue decrease of contraction elicited by the 60 Hz 3 s burst stimulation, as well as no M-wave decrease in amplitude and conduction time. Thus, there was no indication of peripheral muscle fatigue. MF decrease was present in all individuals throughout the fatiguing contraction and it was not correlated with the magnitude of superimposed force. These observations indicate that an absence of superimposed electrically-induced muscle contraction does not allow us to conclude the existence of a sole peripheral muscle fatigue in these circumstances. PMID- 12586517 TI - Continuous, intermitted and sporadic motor unit activity in the trapezius muscle during prolonged computer work. AB - The Cinderella hypothesis postulates the continuous activity of specific motor units (MUs) during low-level muscle contraction. The MUs may become metabolically overloaded, with the subject developing muscle pain and strain. The hypothesis requires MUs that are active for a time long enough to actually damage muscle fibers. The aim of this study was to determine if there are continuously active MUs in the right trapezius muscle during normal computer work using a computer mouse. Fourteen healthy subjects executed an interactive computer-learning program (ErgoLight) for 30 min. Six-channel intramuscular EMG and two-channel surface EMG signals were recorded from two positions of the trapezius muscle. Decomposition was achieved with automated, multi-channel, long-term decomposition software (EMG-LODEC). In two out of the 14 subjects, three MUs were continuously active throughout the 30 min. Although the majority of the MUs were active during only part of the experimental session, an ordered on-off behavior (e.g. substitution) pattern was not observed. As long-lasting activity was verified in some subjects, the results support the Cinderella hypothesis. However, it cannot be concluded here how long the MUs could stay active. If continuous activity overloads low threshold MUs, the potential exists for selective fibre injuries in low threshold MUs of the trapezius muscle in subjects exposed to long-term computer work. PMID- 12586518 TI - Simulation analysis of the ability of different types of multi-electrodes to increase selectivity of detection and to reduce cross-talk. AB - Selectivity of different one- and two-dimensional multi-electrodes and their ability to reduce cross-talk were analyzed. Signals from an individual motor unit (MU) were calculated as a single convolution of intracellular action potential (IAP) first temporal derivative and spatially filtered MU impulse response. It was shown that the uptake area (irrespective of the way it was defined) could not characterize electrode properties reliably because its estimate depended on the source parameters. Due to the different decline of individual phases of MU signals with depth, electrode should provide higher spatial and temporal resolution of the main phases for better selectivity and greater suppression of the terminal phases for cross-talk reduction. A two-dimensional normal double differentiating (NDD) electrode provided almost the same or slightly lower selectivity but weaker reduction of cross-talk than a longitudinal double differentiating (LDD) electrode. A transversal double differentiating (TDD) electrode provided a lower selectivity and weaker reduction of cross-talk than a LDD electrode. A new, BiTDD multi-electrode (performing difference between signals detected by two TDD electrodes) provided the best selectivity and reduction of cross-talk. To obtain the smallest cross-talk, a BiTDD electrode should be positioned above the end-plate region, while LDD, TDD, or NDD electrodes-above the ends of muscle that produced it. Signal differentiation improved selectivity but increased cross-talk. PMID- 12586519 TI - Amplitude and spectral characteristics of biceps Brachii sEMG depend upon speed of isometric force generation. AB - In the present study the influence of speed of contraction on the interplay between recruitment and firing rate of motor units (MUs) was assessed. The surface electromyographic (sEMG) signal was recorded in nine healthy subjects from the right biceps brachii using a linear electrode array during ramp isometric contractions (from 0 to 100% of the maximal voluntary force, MVC) at 5, 10, and 20% MVC s(-1) (ramp phase), followed by 10 s of sustained MVC (hold phase). The median frequency (MDF), Root Mean Square (RMS) and conduction velocity (CV) of sEMG, were computed on adjacent epochs covering a force range of 5% MVC each. Full motor unit recruitment (FMUR) point was assessed as the force level at which MDF reached its maximum value; the MDF decay during the hold phase was taken as an index of localized muscle fatigue. At 5% MVC s(-1), FMUR was reached at 52.3% MVC. At 10%MVC s(-1) FMUR was achieved at 58% MVC; while at 20% MVC s(-1) FMUR point was located at 77% MVC, being statistically different from 5 and 10% MVCs(-1) ramps (p<0.05). The MDF decay was steeper at higher speed. CV modifications mirrored those reported for MDF. The RMS increased in a curvilinear fashion and the maximum value was always attained during the hold phase. Our findings suggest that MU recruitment strategies are significantly related to the speed of contraction even in a single muscle. PMID- 12586520 TI - The effects of precision demands during a low intensity pinching task on muscle activation and load sharing of the fingers. AB - High precision demands in manual tasks can be expected to cause more selective use of a part of the muscular synergy involved. To test this expectation, load sharing of the index finger and middle finger was investigated during a pinching task. Myoelectric activation of lower arm and neck-shoulder muscles was measured to see if overall level of effort was affected by precision demands. Ten healthy female subjects performed pinching tasks with three levels of force and three levels of precision demands. The force level did not significantly affect the relative contribution of the index and middle finger to the force. Higher precision demands, however, led to higher contribution of the index finger to the pinch force. Consequently, a more selective load of the forearm and hand occurs during tasks with high precision demands. The variability of the force contribution of the fingers increased during the task. No effects of precision demand on the activation of forearm and neck-shoulder muscles were found. Force level did affect the EMG parameters of several muscles. The effects were most apparent in the muscles responsible for the pinch force, the forearm muscles. Activation of these muscles was higher at higher force levels. In the trapezius muscle at the dominant side EMG amplitudes were lower at the high pinch force compared to the low force and median force conditions. PMID- 12586521 TI - Knee extensor torque and quadriceps femoris EMG during perceptually-guided isometric contractions. AB - The aim of this study was to examine superficial quadriceps femoris (QF) EMG and torque at perceived voluntary contraction efforts. Thirty subjects (15 males, 15 females) performed 9, 5 s, sub-maximal contractions at prescribed levels of perceived voluntary effort at points 1-9 on an 11-point scale (0-10), in a random order. Surface electromyograms (EMG) of the vastus medialis (VM), vastus lateralis (VL), and rectus femoris (RF) muscles, as well as QF peak torque (PT), average torque (AT), and torque coefficient of variation (C.V.), were sampled. The raw EMG signals were full-wave rectified and integrated over the middle three s of each contraction. The sampled EMG signals, and PT and AT at each perceived exertion level were normalized to the average of three maximal voluntary contractions. The normalized EMG and torque values at each perceived exertion level were then compared to equivalent percent values (i.e., 10% at a perceived level of 1). The results demonstrated that at all perceived exertion levels, with the exception of the RF at a level of 2 which was equivalent to 20%, and the VL and RF muscles at a level 1 in which activation was greater than 10%, activation was significantly less than the equivalent percent value at each point on the scale. VM EMG was found to be less than the VL and RF from contraction levels 3 9. PT was shown to be less than the equivalent percent values at contraction levels 6-9. The AT was found to be lower than the expected percent value at perceived effort levels 2-9. Torque C.V. was not found to be different across the range of perceived effort. The major findings of this study suggested that humans over-estimate voluntary QF muscle torque when guided by perceptual sensations. It is also suggested that the produced EMG signals revealed a reliance on the VL muscle for knee extensor torque generation at sub-maximal levels. PMID- 12586522 TI - The effect of head position, electrode site, movement and smoothing window in the determination of a reliable maximum voluntary activation of the upper trapezius muscle. AB - Quantitative measures derived from raw myoelectric signal (MES) data must be normalized to allow for comparisons both within and between subjects. The most common method of normalization involves dividing the root mean square (RMS) amplitude of the MES recorded during a given activity by the RMS of the MES elicited during a maximal voluntary isometric contraction (MVIC) of that particular muscle. The objective of this study was to use surface-recorded MES amplitude to determine the combination of electrode site, test position, head posture, and smoothing window that elicits the highest and most reliable MES amplitudes during an MVIC of the upper trapezius (UT) muscle. Ten volunteers had surface electrodes positioned at five sites on the UT of their dominant side. Three trials of each of three MVIC test positions were performed both with the head in neutral and rotated 45 degrees to the contralateral side. A repeated measures ANOVA was used for statistical hypothesis testing. Coefficients of variation were used to quantify the between-factor variability introduced in each case. In addition, the data were re-analyzed using moving windows of 100 to 500 ms in length, and an ANOVA was used to determine the effect of window length on both the amplitude and variability of the estimates of maximum voluntary activation (MVE). Head position had no significant effect on RMS amplitude of the MVIC in any of the test positions. There was a significant electrode site by test position interaction effect. Bonferroni post-hoc analyses were performed on this interaction by fixing test position and testing electrode site, revealing that Sites 1 (2 cm lateral to the midpoint between C7 spinous process and the posterolateral border of the acromion) and 4 (2 cm posterior to Site 1) recorded significantly higher RMS values for all test positions, and were not significantly different from each other. Fixing electrode site, the test position analysis revealed that abduction of the humerus, and abduction with external rotation of the humerus produced significantly higher RMS values than shoulder elevation at both Sites 1 and 4, and that abduction produced a significantly higher RMS amplitude than abduction in external rotation at Site 1. The results confirmed that Sites 1 and 4 consistently produced the highest MES amplitudes for all movements. Pure abduction consistently elicited maximal RMS values; however there is concern regarding supraspinatus cross talk during this movement. Site 1 was found to produce the most reliable data. A moving window of 100 ms was found to generate MVE estimates that were significantly higher than windows ranging from 200 ms to 500 ms in length. There was no effect of window length on the reliability of the MVEs. Based on this study, it was concluded that abduction or abduction with the arms in lateral rotation should be used as normalization contraction positions for the upper trapezius muscle. During this movement, Site 1 data smoothed with a moving window of 100 ms produces the highest amplitude MVE data but window lengths greater than 200 ms produce more stable estimates in terms of being able to compare studies in which moving windows are used to compute RMS. PMID- 12586523 TI - Lower extremity muscle activities during cycling are influenced by load and frequency. AB - The purpose of this experiment was to investigate the effects of frequency and inertia on lower extremity muscle activities during cycling. Electromyographic (EMG) data of seven lower extremity muscles were collected. Sixteen subjects cycled at 250 W across different cadences (60, 80, and 100 rpm) with different loads (0, 0.5, 1.0, 1.5, and 2.0 kg) attached to distal end of their thighs. Load and cadence interactions were observed for the offset of the biceps femoris (BF), the active duration of the rectus femoris (RF), and the peak magnitudes of the vastus lateralis (VL) and the tibialis anterior (TA). Cadence effects were observed in the onset of the gluteus maximus (GM), RF, BF, VL, and TA; the offset of the GM, RF, BF, VL; the duration of the BF and TA; the peak magnitude of the RF and gastrocnemius (GAS); and the crank angle at which the peak magnitude was achieved of the BF, GAS, and soleus (SOL). Load effect was observed from the onset of RF and SOL, the offset of RF, the duration of SOL, and the peak magnitude of BF. These results indicate that inertial properties influence the lower extremity muscular activity in addition to the cadence effect. PMID- 12586524 TI - Quantification of concentric-needle-induced insertional activity by turn/amplitude analysis. AB - Quantification of needle-induced insertional activity (IA) by means of the turn/amplitude analysis (TAA) has not been attempted so far. IA was recorded from the right brachial biceps and right anterior tibial muscle of 29 healthy subjects, 17 woman, 12 men, aged 24-79 years and 51 patients with neuromuscular disorders (29 neuropathies and 22 myopathies), 24 woman, 27 men, aged 17-81 years. IA was analyzed with regard to IA duration (IAD), turns/second (T/S), amplitude/turn (A/T), T/S:IAD and A/T:IAD. IAD was not significantly different between healthy and diseased subjects in either of the two investigated muscles. On the contrary, T/S, A/T, T/S:IAD and A/T:IAD were significantly increased in the brachial biceps and anterior tibial muscle of patients with neuropathies. In myopathies T/S were increased in both muscles and the T/S:IAD in the anterior tibial muscle. Quantification of IA by TAA increases the sensitivity of conventional EMG from 38 to 69% in neuropathies and from 41 to 73% in myopathies. It is concluded that quantification of IA by TAA is of diagnostic support in the detection of neuromuscular disorders. The most useful parameter in this respect is the T/S, which, contrary to IAD, is increased in patients with neuropathies as well as patients with myopathies. PMID- 12586525 TI - Electromyographic responses to prescribed mastication. AB - The aim was to understand between-volunteer differences in Electromyography (EMG) behaviour during chewing. EMG was used to record the electrical activity of the temporal and masseter muscles of volunteers, who carried out mastication movements by operating calibrated springs held between their incisors. The volunteers coordinated their jaw movements with the signal produced by a metronome, at four rates: 30, 60, 90 and 120 beats per minute (bpm). Raw data were analyzed to examine the distributions of the intervals between chews. For the highest prescribed chew rates, the volunteers' distributions were very similar. The distributions varied most for the 30 bpm data, suggesting that volunteers differed in their ability to carry out and maintain this prescribed chewing pattern. The data were Fourier transformed to give power spectra in the frequency domain. The low frequency (<10 Hz) region contained spectral features related to the prescribed chew rate. Principal component analysis of the power spectra revealed that readings from each volunteer clustered together, and the clusters could be largely separated. Such grouping was found irrespective of whether data from each chew rate were analyzed separately or simultaneously. This indicated that within-volunteer variance, arising from the different chew rates as well as between-session variance, is lower than between-volunteer variance; even when individuals are asked to make jaw movements in the same prescribed manner, they can nevertheless be uniquely distinguished by their muscle activity as recorded by EMG. PMID- 12586527 TI - Sleep apnoea syndrome today: much done, more to do. PMID- 12586528 TI - The upper airway in sleep: physiology of the pharynx. AB - The upper airway is the primary conduit for passage of air into the lungs. Its physiology has been the subject of intensive study: both passive mechanical and active neural influences contribute to its patency and collapsibility. Different models can be used to explain behavior of the upper airway, including the "balance of forces" (airway suction pressure during inspiration versus upper airway dilator tone) and the Starling resistor mechanical model. As sleep is the primary state change responsible for sleep disordered breathing (SDB) and the obstructive apnea/hypopnea syndrome (OSAHS), understanding its effects on the upper airway is critical. These include changes in upper airway muscle dilator activity and associated changes in mechanics and reflex activity of the muscles. Currently SDB is thought to result from a combination of anatomical upper airway predisposition and changes in neural activation mechanisms intrinsic to sleep. Detection of SDB is based on identifying abnormal (high resistance) breaths and events, but the clinical tools used to detect these events and an understanding of their impact on symptoms is still evolving. Outcomes research to define which events are most important, and a better understanding of how events lead to physiologic consequences of the syndrome, including excessive daytime somnolence (EDS), will allow physiologic testing to objectively differentiate between "normal" subjects and those with disease. PMID- 12586529 TI - Obstructive sleep apnoea syndrome--an oxidative stress disorder. AB - Obstructive sleep apnoea syndrome (OSA) is associated with increased cardiovascular morbidity and mortality. However, the underlying mechanisms are not entirely understood. This review will summarize the evidence that substantiates the notion that the repeated apnoea-related hypoxic events in OSA, similarly to hypoxia/reperfusion injury, initiate oxidative stress. Thus, affecting energy metabolism, redox-sensitive gene expression, and expression of adhesion molecules. A limited number of studies substantiate this hypothesis directly by demonstrating increased free radical production in OSA leukocytes and increased plasma-lipid peroxidation. A great number of studies, however, support this hypothesis indirectly. Increase in circulating levels of adenosine and urinary uric acid in OSA are implicated with increased production of reactive oxygen species (ROS). Activation of redox-sensitive gene expression is suggested by the increase in some protein products of these genes, including VEGF, erythropoietin, endothelin-1, inflammatory cytokines and adhesion molecules. These implicate the participation of redox-sensitive transcription factors as HIF 1 AP-1 and NFkappaB. Finally, adhesion molecule-dependent increased avidity of OSA monocytes to endothelial cells, combined with diminished NO bioavailability, lead to exaggerated endothelial cell damage and dysfunction. Cumulatively, these processes may exacerbate atherogenic sequelae in OSA. PMID- 12586530 TI - Home diagnosis of the obstructive sleep apnoea/hypopnoea syndrome. AB - Polysomnography has been accepted by many as a "gold standard" for the diagnosis of the Obstructive Sleep Apnoea/Hypopnoea Syndrome (OSAHS). Although polysomnography is a good method for diagnosing OSAHS, there is no evidence that the results of polysomnography more accurately identify patients with the syndrome than more simple investigations which may be done at lower cost in the patient's home. This article examines the evidence for and against home sleep studies and concludes that home sleep studies have a role. Precisely what that role is will depend on financial and organisational aspects for each sleep centre. PMID- 12586531 TI - Treatment of obstructive sleep apnea in children: do we really know how? AB - Obstructive sleep apnea syndrome (OSAS) is a frequent, albeit underdiagnosed problem in children. If left untreated, OSAS may lead to substantial morbidities affecting multiple target organs and systems. The immediate consequences of OSAS in children include behavioral disturbance and learning deficits, pulmonary hypertension, as well as compromised somatic growth. However, if not treated promptly and early in the course of the disease, OSAS may also impose long-term adverse effects on neurocognitive and cardiovascular function, thereby providing a strong rationale for effective treatment of this condition. This review provides a detailed description of the current treatment modalities for pediatric OSAS, and uncovers the potential limitations of the available data on these issues. Furthermore, we postulate that OSAS will persist relatively often after tonsillectomy and adenoidectomy, and that critical studies need to be conducted to identify such patients and refine the clinical management algorithm for pediatric OSAS. PMID- 12586532 TI - Improving CPAP use by patients with the sleep apnoea/hypopnoea syndrome (SAHS). AB - Though continuous positive airway pressure (CPAP) is the treatment of choice for the sleep apnoea/hypopnoea syndrome (SAHS), suboptimal adherence to CPAP is common. Internationally, some 5-50% of SAHS patients recommended for CPAP either reject this treatment option or discontinue within the first week, and 12-25% of remaining patients can be expected to have discontinued CPAP by 3 years. Biomedical investigations of patients' CPAP use reveal frequent adverse effects, weak prospective relationships between symptomatic or physiological disease severity and CPAP use, and moderate correlations between use and benefit. Relatively expensive high-technological interventions to improve CPAP use (e.g. "intelligent" CPAP, humidification) are the subject of several well-conducted studies favouring their effectiveness. More basic educational and behavioural supports, and low-technological interventions (e.g. chinstraps, mask re-fitting) appear valued, but are currently less rigorously evaluated. In other diseases with demanding treatment regimens, cognitive constructs including health attitudes and beliefs (health value, locus of control, chance, powerful others, self-efficacy) and mental and physical health status are significant predictors of adherence. The enhancement of multidisciplinary models with psychosocial interpretations may provide increased explanatory and interventional potential in models of CPAP use. While acknowledging the scarcity of evidence, a structured, multidisciplinary, cost-efficient model is suggested, containing educational, behavioural and technological components as basic support, and with high expertise cognitive-behavioural intervention in more difficult cases of low CPAP use. PMID- 12586533 TI - On the role of serotonin in sleep regulation and depression. A commentary on "Neurobiological bases for the relation between sleep and depression" (J. Adrien). PMID- 12586535 TI - Effect of amidine derivatives on nitric oxide production by Leishmania amazonensis promastigotes and axenic amastigotes. AB - The effects of pentamidine isethionate (reference drug) and N,N'-diphenyl-4 methoxy-benzamidine (test compound) on NO. production by Leishmania amazonensis promastigotes and axenic amastigotes were investigated by measuring nitrite, a by product of nitric oxide released into culture supernatants. The NO. production by infective promastigotes was inhibited by OCH(3)-amidine in about 23.53% and by pentamidine in only 3.78%. In axenic amastigotes, the inhibition of NO. production by OCH(3)-amidine was significantly higher (52.94%; p=0.01) than that by pentamidine, which inhibited this radical production nonsignificantly (25.29%; p=0.1). The mechanism of amidine derivatives, as an antimicrobial agent, is unknown. However, other amidines, such as a diamidine (pentamidine), contain chemical structures shared by the guanidino group of the nitric oxide synthase substrate L-arginine, suggesting the possibility of an interaction with this enzyme or electronic factors (substituent constant) that alter physical and chemical properties significant for biological activity. PMID- 12586536 TI - Functional characterization of Glu298Asp mutant human endothelial nitric oxide synthase purified from a yeast expression system. AB - The Glu298Asp polymorphism of human endothelial nitric oxide synthase (eNOS) has been reported to be associated with several cardiovascular diseases, including hypertension and myocardial infarction. Therefore, we investigated the effect of the Glu298Asp (E298D) mutation on the function of purified recombinant eNOS expressed in the yeast Pichia pastoris. Wild type (WT) and mutant exhibited comparable affinities for L-arginine (K(m) values 4.4+/-0.6 and 5.2+/-0.8 microM, respectively) and V(max) values (142+/-36 and 159+/-29 nmol of L-citrulline/mg min, respectively). The E298D mutation affected neither electron transfer through the reductase domain (measured as cytochrome c reduction) nor reductive O(2) activation (measured either as NADPH oxidation or as H(2)O(2) formation in the absence of L-arginine and tetrahydrobiopterin (BH4)). The mutant was activated by BH4 with an EC(50) of 0.24+/-0.04 microM, a value comparable to that obtained with WT eNOS (0.22+/-0.02 microM). Activation of the enzyme by Ca(2+) was not affected (EC(50)=0.50+/-0.04 and 0.49+/-0.02 microM for WT and E298D eNOS, respectively). Calmodulin (CaM) affinity, studied by radioligand binding using 125I-labeled CaM, revealed virtually identical K(D) (3.2+/-0.5 and 4.0+/-0.3nM) and B(max) (1.4+/-0.2 and 1.2+/-0.3 pmol/pmol subunit) values for WT and E298D eNOS, respectively. Furthermore, E298D eNOS did not differ from the WT enzyme with respect to heme and flavin content or the ability to form SDS-resistant dimers. To summarize, we obtained no evidence for altered enzyme function of the eNOS mutant that could explain endothelial dysfunction associated with the E298D polymorphism. PMID- 12586537 TI - Mechanism of vasculitis and aneurysms in Kawasaki disease: role of nitric oxide. AB - NO in vivo has both beneficial and nonbeneficial effects depending on site and concentration. Peroxynitrite, resulting from the reaction of NO with superoxide radical, causes cellular damage. Nitrotyrosine, end product of NO's toxic effects on cellular proteins, is a stable compound that can be used to detect evidence of harmful quantities of NO. We sought to detect nitrotyrosine in coronary arterioles of DBA/2 mice injected intraperitoneally with Lactobacillus casei cell wall. The inflammatory response induced occurred in perivascular fashion and involved mainly macrophages. It was variable according to time points, being severe on days 10 and 14 and mild to moderate on days 3 and 7. Few basal inflammatory cells appeared in controls injected with phosphate-buffered saline. Western immunoblots of homogenized hearts on days 10 and 14 demonstrated specific nitrated proteins. Immunohistochemistry of frozen sections of diseased hearts showed positive immunoreactivity for nitrotyrosine in coronary arterioles at the same time points. These findings were absent in the controls. We also determined the expression of inducible nitric oxide synthase (iNOS) in controls on days 10 and 14. iNOS colocalized with nitrotyrosine in perivascular macrophages and coronary arterioles of treated mice. Additionally, aneurysms were found on day 10 and intracardiac hemorrhage with consequent death on day 14. These observations supply evidence that NO through its reactive product, peroxynitrite, and its antigen/tissue marker, nitrotyrosine, is directly involved in coronary arteritis and aneurysm development in mice models of Kawasaki disease (KD). This article shows that macrophages are central to this and bolsters the likelihood of L. casei being the cause of KD. PMID- 12586538 TI - Nitrate in exhaled breath condensate of patients with different airway diseases. AB - There is an increasing interest in the measurement of nitric oxide (NO.) in the airways. NO. is a free radical that reacts rapidly with reactive oxygen species in aqueous solution to form peroxynitrite which can then break down to nitrite (NO(2)(-)) and nitrate (NO(3)(-)). NO(3)(-) is considered a stable oxidative end product of NO. metabolism. The aim of this study was to assay NO(3)(-) in exhaled breath condensate (EBC) of normal nonsmoking and smoking subjects, asthmatics, patients with obstructive pulmonary disease (COPD), and patients with community acquired pneumonia (CAP). EBC was collected using a glass condenser and samples were assayed for NO(3)(-) by ion chromatography followed by conductivity measurement. NO(3)(-) was detectable in EBC of all subjects. NO(3)(-) was elevated in smokers [median (range)] [62.5 (9.6-158.0) microM] and in asthmatics [68.0 (25.8-194.6) microM] compared to controls [9.6 (2.6-119.4) microM; p=0.003 and p=0.006, respectively], whereas NO(3)(-) was not elevated in COPD patients [24.1 (1.9-337.0 microM]. The concentration of NO(3)(-) in patients with CAP [243.4 (26.1-584.5) microM] was higher than that in controls (p=0.002) and NO(3)( ) values decreased after treatment and recovery from illness [40.0 (4.1-167.0) microM, p=0.009]. This study shows that NO(3)(-) is detectable in EBC of healthy subjects and it varies in patients with inflammatory airway diseases. PMID- 12586539 TI - Citrulline immunohistochemistry may not necessarily identify nitric oxide synthase activity: the pitfall of peptidylarginine deiminase. AB - Nitric oxide synthase (NOS) converts L-arginine as a substrate to form nitric oxide and the "by-product" citrulline. To characterize NOS activity in the nervous tissue at the single-cell level, citrulline immunostaining is considered to be a suitable means of working on the principle that in brain tissue, due to the incomplete urea cycle, citrulline is produced exclusively by NOS. This assumption is correct for free citrulline but it does not consider the conversion of arginine to citrulline residues of proteins by the calcium-dependent peptidylarginine deiminase (PAD). Using a polyclonal antiserum against citrulline we observed in cerebellar cell cultures immunopositivity in a few, mostly NOS positive, neurons, in activated microglia, and in oligodendroglia (which under control conditions are in doubt to be able to express NOS), but not in astroglia. Treatment with the excitotoxin kainate substantially enhanced the staining intensity for citrulline in neurons and glial cells. To distinguish between free (NOS-related) and protein-bound (PAD-related) citrulline we blocked NOS activity by 7-nitroindazole or L-N5-(1-iminoethyl)lysine. The results provide evidence that citrulline immunolabeling results partly from PAD-mediated protein citrullination, enhanced pathophysiologically under stimulated conditions by exposure to kainate. Our immunocytochemical observations were corroborated by Western blot analysis showing several bands of citrulline-positive proteins, whose number and staining intensity depended on kainate treatment and calcium ions. PMID- 12586540 TI - Cytosolic and mitochondrial glutathione in microglial cells are differentially affected by oxidative/nitrosative stress. AB - Glutathione (GSH), the major cellular protectant against reactive oxygen and nitrogen species, is compartmentalized in a cytosolic (c) and a mitochondrial (mt) pool. We investigated how c-GSH and mt-GSH are differentially affected by endogenously produced nitric oxide (NO). Microglial cell line (N9) cultures were immunostimulated with lipopolysaccharide/interferon-gamma to elicit the inducible isoform of NO synthase (iNOS). Despite a significant reduction in total GSH, the mt-GSH remained nearly unaffected by iNOS-mediated NO production. To investigate possible consequences of GSH depletion on the mitochondrial membrane potential, we used buthionine sulfoximine (BSO) to reduce separately the c-GSH, whereas ethacrynic acid (EA) was applied to deplete both mt-GSH and c-GSH. The mitochondrial membrane potential was more vulnerable to NO exposure in EA pretreated cultures than in BSO-pretreated cultures, indicated by a potentiated release of tetramethylrhodamine from mitochondria into the cytosol. To relate the EA-mediated decrease in mitochondrial membrane potential to the oxidant buildup after GSH depletion, we loaded the cells with the oxidant-sensitive fluorochrome 2',7'-dihydrodichlorofluorescein (DCF) diacetate. EA treatment caused an increase in DCF fluorescence over time that was potentiated when the iNOS expression was stimulated. Inhibition of NO production abolished this effect. We conclude that endogenous NO production in microglial cells does not compromise the mt-GSH pool which, in turn, might explain the ability of these cells to combat high-output NO production. PMID- 12586541 TI - Nitric oxide induces muscular relaxation via cyclic GMP-dependent and independent mechanisms in the longitudinal muscle of the mouse duodenum. AB - The aim of this study was to investigate, in mouse duodenum, the role of nitric oxide (NO) in the relaxation of longitudinal muscle evoked by nerve activation and the coupled action mechanism. Electrical field stimulation (EFS; 0.5 ms, 10-s train duration, supramaximal voltage, at various frequencies) under nonadrenergic noncholinergic conditions evoked muscular relaxation occasionally followed, at the higher stimulus frequencies, by rebound contractions. Inhibition of the synthesis of NO by N(omega)-nitro-L-arginine methyl ester (L-NAME; 100 microM) virtually abolished the evoked relaxation. The relaxation was reduced also by apamin (0.1 microM) and by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 1 microM), a guanylyl cyclase inhibitor. The coadministration of apamin and ODQ produced additive effects on the responses to EFS. Sodium nitroprusside (0.1-100 microM) produced a concentration-dependent reduction of the phasic spontaneous activity and at the highest dose used suppressed phasic activity and induced muscular relaxation. These effects were tetrodotoxin and L-NAME resistant and were antagonized both by apamin and by ODQ. 8-Bromoguanosine 3',5'-cyclic monophosphate (0.1-100 microM) reduced in a concentration-dependent manner the spontaneous mechanical activity and at 100 microM suppressed the phasic activity and induced muscular relaxation, not antagonized by apamin. This study indicates that NO is the primary transmitter released by inhibitory nerves supplying the longitudinal muscle of mouse duodenum and that guanylate cyclase stimulation and opening of Ca(2+)-dependent K(+) channels are independent mechanisms working in parallel to mediate NO action. PMID- 12586542 TI - Long-term dietary L-arginine supplementation attenuates proteinuria and focal glomerulosclerosis in experimental chronic renal transplant failure. AB - Glomerular endothelial nitric oxide synthase expression is decreased in humans during acute rejection and chronic renal transplant failure (CRTF). This may contribute to vascular damage through changes in the renal hemodynamics and enhanced endothelial adhesion of leukocytes and platelets. Dietary supplementation of L-arginine may increase endothelial NO production, thereby protecting the vascular wall and improving renal hemodynamics. We tested the hypothesis that long-term L-arginine supplementation attenuates the development of CRTF in an experimental model for renal transplantation. In the Fisher 344 to Lewis rat model for renal transplantation, renal function and histology of untreated rats was compared with rats receiving L-arginine in the drinking water (10g/L), starting 2 days before transplantation. Every 4 weeks systolic blood pressure was measured and serum and urine were collected for measurement of nitrite and nitrate (NO(x)), creatinine, and proteinuria. At 34 weeks the histological renal damage was assessed by scoring focal glomerulosclerosis and measurement of alpha-smooth muscle actin (alpha-SMA) expression. Urinary NO(x) was significantly increased in treated animals. Proteinuria was significantly lower in L-arginine-treated animals from week 24 onward (p<0.05). Plasma creatinine and creatinine clearance did not differ between the groups. The focal and segmental glomerulosclerosis (FGS) score (max 400) at week 34 was also significantly lower in treated rats arbitrary U (20+/-21 vs 61+/-67 arbitrary U; p<0.05). The expression of alpha-SMA was lower in L-arginine-treated rats than in untreated rats (1.93+/-0.8% area surface vs 3.64+/-2.5% area surface). In conclusion, in this experimental model for CRTF, L-arginine administration significantly reduced FGS and proteinuria, without affecting renal function. Our data suggest that dietary L-arginine supplementation attenuates progression of CRTF and may therefore be an additional therapeutic option in human renal allograft recipients. PMID- 12586543 TI - Nitric oxide donor increases the efficiency of cytostatic therapy and retards the development of drug resistance. AB - The potentiality to increase the chemotherapeutic effectiveness of some cytostatics in low, subtherapeutic doses in combination with nitric oxide (NO) donor has been shown. This type of combined therapy results in significant increase in life span and number of survivors among mice bearing leukemias P388 and L-1210. A similar effect was observed for intracerebral leukemia P388 transplantation. In this case the life span of mice treated with cyclophosphamide and NO donor increased by three times in comparison to therapy with cyclophosphamide alone. The coinjection of nitric oxide donor and cytostatics improved the antimetastatic activity of the cytostatics: the index of melanoma B16 metastasis inhibition at the cyclophosphamide monotherapy is 50%; on addition of NO donor the index is over 80%. Comparative studies of NO donor (organic nitrate) and a similar compound in which ONO(2) moieties were replaced by OH groups demonstrated that the presence of NO(2) is required for adjuvant activity of compounds and confirmed that nitric oxide modifies the antitumor effects of cytostatics. It is shown also that nitric oxide donor retards the development of drug resistance to cyclophosphamide. PMID- 12586544 TI - Regulation of NF-kappaB activation by protein phosphatase 2B and NO, via protein kinase A activity, in human monocytes. AB - It has been reported previously that a short synthetic immunomodulating peptide (Pa) and the neuropeptide beta-endorphin modulate the immune system. We have found now that NF-kappaB participates in the stimulation of monocytes by both peptides and we investigated the molecular mechanism by which these stimuli activate NF-kappaB. Pa and beta-endorphin induce accumulation of cyclic 3('),5(') adenosine monophosphate (cAMP) in a calcium/calmodulin-dependent fashion since it was completely inhibited by the calmodulin antagonist W-7. The effect of these complexes seems to be mediated, at least in part, by nitric oxide (NO) synthesized by constitutive NO synthase since the NO synthase inhibitor N-methyl L-arginine (NMLA) reduced the elevation of cAMP. Furthermore, the NO donor SIN-1 provoked nitration of G(S)alpha, leading to the cAMP elevation that was suppressed by the G(S)alpha-selective antagonist NF-449. Interestingly, the rapid degradation of NF-kappaB inhibitor IkappaBalpha induced by Pa- and beta-endorphin was reversed by a pretreatment with H-89 and cyclosporin A, inhibitors of protein kinase A (PKA) and protein phosphatase 2B (PP2B), respectively. These observations are consistent with the inhibition caused by W-7, NMLA, H-89, and cyclosporin A on NF-kappaB induction by these agonists, indicating the involvement of PKA and PP2B in the regulation of NF-kappaB in human monocytes. PMID- 12586545 TI - Inhibition of macrophage nitric oxide production by gangliosides derived from a spontaneous T cell lymphoma: the involved mechanisms. AB - Gangliosides (DLG) derived from a spontaneous T cell lymphoma (Dalton's lymphoma) have been shown to impair the ability of lipopolysaccharide-activated macrophages to produce nitric oxide (NO). However, the mechanism and nature of this effect is not known. In this investigation, we sought to (1) determine whether the inhibitory action of DLG on macrophages is through the modulation of inducible nitric oxide synthase (iNOS) expression and (2) identify the possible mechanisms and signal transduction events underlying the inhibitory action of DLG. Immunoblot analysis of DLG-treated macrophages showed a decrease in iNOS expression. DLG also inhibited the production of monokines interleukin-1 and tumor necrosis factor by macrophages. However, the DLG-induced inhibition was reversible in nature. Studies showed that DLG-induced inhibition of macrophage activation could be blocked by sodium orthovanadate, indicating a role of phosphatase activity in ganglioside-induced inhibition. PMID- 12586546 TI - Neurotoxicity of pneumolysin, a major pneumococcal virulence factor, involves calcium influx and depends on activation of p38 mitogen-activated protein kinase. AB - Neuronal injury in bacterial meningitis is caused by the interplay of host inflammatory responses and direct bacterial toxicity. We investigated the mechanisms by which pneumolysin, a cytosolic pneumococcal protein, induces damage to neurons. The toxicity after exposure of human SH-SY5Y neuroblastoma cells and hippocampal organotypic cultures to pneumolysin was time- and dose-dependent. Pneumolysin led to a strong calcium influx apparently mediated by pores on the cell membrane formed by the toxin itself and not by voltage-gated calcium channels. Buffering of intracellular calcium with BAPTA-AM [1, 2-bis (o aminophenoxy) ethane N, N, N', N'-tetraacetic acid tetra(acetomethoxyl) ester] improved survival of neuronal cells following challenge with pneumolysin. Western blotting revealed increased phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) as early as 30 min after challenge with pneumolysin. SB 203580, a potent and selective inhibitor of p38 MAPK, rescued human neuronal cells from pneumolysin-induced death. Inhibition of the mitochondrial permeability transition pore using bongkrekate and caspase inhibition also improved survival following challenge with the toxin. Modulation of cell death pathways activated by pneumolysin may influence the outcome of pneumococcal meningitis. PMID- 12586547 TI - Differential changes in striatal projection neurons in R6/2 transgenic mice for Huntington's disease. AB - In early adult-onset Huntington's disease (HD), enkephalinergic striatopallidal projection neurons show preferential loss, reduced preproenkephalin (PPE) expression in surviving striatopallidal neurons, and loss of fibers in their projection target area. We examined PPE and PPT (preprotachykinin) gene expression in striatal projection neurons and in striatal projection fibers immunoreactive for the PPE product enkephalin (ENK) and the PPT product substance P (SP) in a transgenic HD model, the R6/2 mouse, to see if changes occur in these neuron types similar to those seen in early adult-onset HD. The results show that PPE mRNA level, the number of striatal neurons expressing PPE, and the staining intensity of fibers immunoreactive for ENK in the pallidum were all decreased. By contrast, the SP-containing striatal projection systems to the pallidum and substantia nigra were relatively normal in R6/2 mice. The selective reduction in striatal PPE in R6/2 mice is reminiscent of adult-onset HD, but the preservation of the striatonigral projection system is not. Thus, R6/2 mice do not strictly mimic adult-onset HD in their striatal pathology. PMID- 12586548 TI - Astrocytes accumulate 4-hydroxynonenal adducts in murine scrapie and human Creutzfeldt-Jakob disease. AB - Scrapie-infected mice are considered a model for study in prion diseases, which are characterized by the progressive accumulation in the brain of an abnormal isoform (PrPsc) of the normal cellular prion protein (PrPc). Increasing data suggest that the neurodegenerative process in prion diseases may result, at least partially, from a defect in antioxidant function, but so far in vivo oxidative stress remains poorly documented. We report here that 4-hydroxynonenal, a lipid peroxidation by-product, forms protein adducts in brains of scrapie-infected mice and of Creutzfeldt-Jakob disease affected patients. In scrapie mice, studies on the progression of PrPsc accumulation, glial activation, ubiquitin deposition, and 4-HNE adduct formation allowed us to conclude the late occurrence of oxidative damage in the course of the disease. Massive 4-HNE accumulation was identified in astrocytes, but not in neurons or microglial cells. These findings suggest an important oxidative stress (and subsequent lipid peroxidation) in astrocytes, with possible consequences on their neuronal trophic function. PMID- 12586549 TI - Long-term potentiation is increased in the CA1 area of the hippocampus of APP(swe/ind) CRND8 mice. AB - The present study reports changes in synaptic function and plasticity [long-term potentiation (LTP)] in a recently developed mouse model of Alzheimer's disease (CRND8 line) harboring a double amyloid precursor protein mutation (APP(swe/ind)). In 9-week-old preplaque transgenic (Tg) mice brain slices, basal synaptic function in the hippocampal CA1 area was unchanged. Only one of three different LTP induction protocols revealed early influence of genotype on synaptic plasticity. By 20 weeks of age, there were numerous plaques in the hippocampus from Tg mice associated with more robust evidence for genotype related effects in synaptic function. Field potential maximum slope was consistently decreased and LTP was increased, irrespective of the stimulation protocol used. In addition, there was clear evidence of increased synaptic excitability in Tg mice. Furthermore, the maximum amplitude of evoked IPSCs was decreased whereas the maximum amplitude of evoked EPSCs was increased in 20-week old Tg mice. Collectively, these results suggest a number of APP genotype-related changes in the fine-tuning of the CA1 area circuitry, some of which are likely to contribute to the pathology-dependent effects on LTP observed in CRND8 mice. PMID- 12586551 TI - Impaired eyeblink conditioning and decreased hippocampal volume in PDAPP V717F mice. AB - We examined heterozygous transgenic (Tg) mice that overexpress V717F amyloid precursor protein (APP) for delay eyeblink conditioning (EBC) and hippocampal volume with magnetic resonance imaging (MRI). Platelet-derived APP mice were significantly impaired on EBC relative to wild type (WT) litter-mate controls. T2 weighted spin echo images (62.5 x 125 x 500 microm) of the same mice were acquired under anesthesia using a 9.4T magnet. Tg mice had hippocampal to brain volume ratios that were significantly smaller than WT controls (31% smaller in the rostral dorsal hippocampus, 13-22% smaller among equal dorsal-ventral thirds of a caudal section). These results indicate that overexpression of APP or beta amyloid profoundly affects learning and memory and hippocampal volume. The results also indicate that eyeblink conditioning and quantitative MRI in mice may be useful assays to follow the progression of disease-related changes, and to test the effectiveness of potential therapeutics against Alzheimer's disease. PMID- 12586550 TI - Huntington's disease of the endocrine pancreas: insulin deficiency and diabetes mellitus due to impaired insulin gene expression. AB - In a transgenic mouse model of the neurodegenerative disorder Huntington's disease (HD), age-dependent neurologic defects are accompanied by progressive alterations in glucose tolerance that culminate in the development of diabetes mellitus and insulin deficiency. Pancreatic islets from HD transgenic mice express reduced levels of the pancreatic islet hormones insulin, somatostatin, and glucagon and exhibit intrinsic defects in insulin production. Intranuclear inclusions accumulate with aging in transgenic pancreatic islets, concomitant with the decline in glucose tolerance. HD transgenic mice develop an age dependent reduction of insulin mRNA expression and diminished expression of key regulators of insulin gene transcription, including the pancreatic homeoprotein PDX-1, E2A proteins, and the coactivators CBP and p300. Disrupted expression of a subset of transcription factors in pancreatic beta cells by a polyglutamine expansion tract in the huntingtin protein selectively impairs insulin gene expression to result in insulin deficiency and diabetes. Selective dysregulation of gene expression in triplet repeat disorders provides a mechanism for pleiotropic cellular dysfunction that restricts the toxicity of ubiquitously expressed proteins to highly specialized subpopulations of cells. PMID- 12586552 TI - Evidence for an altered tryptophan metabolism in fibromyalgia. AB - Fibromyalgia (FM) is a prevalent syndrome with chronic pain and a hypothesized underlying disturbance of the tryptophan (TRP) metabolism. We performed a tryptophan depletion (TD) test in 17 FM patients and 17 controls. TRP, 5 hydroxyindoleacetic acid (5-HIAA), kynurenine (KYN), and interleukin-6 (IL-6) were measured. Additionally pain perception was monitored in the FM patients. FM patients and controls exhibited a decrease of TRP and KYN during TD. 5-HIAA levels also decreased in all controls and in 11 FM patients, but showed a marked increase in 6 FM patients. IL-6 significantly increased during TD in the patients, but not in the controls. Pain perception was not affected in the FM patients. These data demonstrate an altered TRP metabolism in a subgroup of FM patients, where the TD seems to activate 5-HT metabolism. Our findings may have diagnostic as well as therapeutic implications in the field of fibromyalgia. PMID- 12586553 TI - Activation of ERK and Akt signaling in focal cerebral ischemia: modulation by TGF alpha and involvement of NMDA receptor. AB - Cerebral ischemia activates ERK and Akt pathways. We studied whether these activations were affected by treatment with the protective growth factor transforming growth factor-alpha (TGF-alpha), and whether they were mediated through N-methyl D-aspartate (NMDA) receptors. The middle cerebral artery was occluded in rats and signaling was studied 1 h later. Noncompetitive NMDA receptor antagonist MK-801 was injected i.p. before the occlusion, whereas in other rats TGF-alpha was given intraventricularly before and after occlusion. Ischemia caused ERK phosphorylation in the nucleus, localized in the endothelium and neurons. Phosphorylation of ERK was prevented by TGF-alpha, but it was enhanced in the nucleus and cytoplasm by MK-801. Also, MK-801 but not TGF-alpha increased p-Akt. Results suggest that preventing ERK activation is related to the protective effect of TGF-alpha, whereas the protective effect of MK-801 is associated with activation of pro-survival Akt. While results support that NMDA receptor signaling precludes Akt activation, we did not find evidence to support that it underlies ischemia-induced ERK phosphorylation. This study illustrates that neuroprotection results from a fine balance between death and survival signaling pathways. PMID- 12586554 TI - Progesterone neuroprotection in the Wobbler mouse, a genetic model of spinal cord motor neuron disease. AB - Motor neuron degeneration characterizes the spinal cord of patients with amyotrophic lateral sclerosis and the Wobbler mouse mutant. Considering that progesterone (PROG) provides neuroprotection in experimental ischemia and injury, its potential role in neurodegeneration was studied in the murine model. Two month-old symptomatic Wobbler mice were left untreated or received sc a 20-mg PROG implant for 15 days. Both light and electron microscopy of Wobbler mice spinal cord showed severely affected motor neurons with profuse cytoplasmic vacuolation of the endoplasmic reticulum and/or Golgi apparatus and ruptured mitochondria with damaged cristae, a profile indicative of a type II cytoplasmic form of cell death. In contrast to untreated mice, neuropathology was less severe in Wobbler mice receiving PROG; including a reduction of vacuolation and of the number of vacuolated cells and better conservation of the mitochondrial ultrastructure. In biochemical studies, we determined the mRNA for the alpha3 subunit of Na,K-ATPase, a neuronal enzyme controlling ion fluxes, neurotransmission, membrane potential, and nutrient uptake. In untreated Wobbler mice, mRNA levels in motor neurons were reduced by half compared to controls, whereas PROG treatment of Wobbler mice restored the expression of alpha3 subunit Na,K-ATPase mRNA. Therefore, PROG was able to rescue motor neurons from degeneration, based on recovery of histopathological abnormalities and of mRNA levels of the sodium pump. However, because the gene mutation in Wobbler mice is still unknown, further studies are needed to unveil the action of PROG and the mechanism of neuronal death in this genetic model of neurodegeneration. PMID- 12586555 TI - A new possible mechanism of human immunodeficiency virus type 1 infection of neural cells. AB - To study the mechanism by which HIV-1 infects neurons we have used human neuroblastoma cell lines (NB). NB (SK-N-SH and SK-N-MC) were found to be susceptible to productive infection by X4 or R5 HIV-1, as detected by viral load and Ag-p24. To identify the putative receptor, we tested the cell surface expression of previously described receptors such as CD4, nucleolin, galactosylceramide, and CCR1, CCR5, and CXCR4 by cytometry and RT-PCR. NB express no CD4 and low levels of galactosylceramide or nucleolin. Furthermore, antibodies to any of these molecules did not affect NB infection. NB express variable levels of CCR5, CCR1, and CXCR4. Interestingly, exogenous heparan sulfate alone was able to substantially inhibit HIV-1 infection, an effect which was potentiated by RANTES or SDF-1 in the HIV-1-infection with R5 or X4 isolates. Besides, anti-CCR5 and anti-CXCR4 significantly blocked HIV-1 infection of R5 and X4 isolates. Our results suggest that HIV-1 entry involves a chemokine-receptor-dependent but CD4 independent entry in neural cells. PMID- 12586557 TI - Evidence for exercise therapy in mechanical neck disorders. AB - In spite of neck disorders being so common in the population, little evidence supporting effective interventions has been identified. The objective of this systematic review was to determine if various exercise methods are effective in treating the different mechanical neck disorders occurring in adults. Sixteen trials were included: nine randomized controlled trials (RCTs) and seven randomized comparative trials (CTs). The average PEDro score indicated moderate methodological quality. PEDro results showed the subject- and therapist-blinding criteria to be inappropriate. Findings revealed relatively strong evidence supporting the effectiveness of proprioceptive exercises and dynamic resisted strengthening exercises of the neck-shoulder musculature for chronic or frequent neck disorders. Moderate evidence was found to support early mobilizing exercises in acute whiplash patients. The evidence identified could not support the effectiveness of group exercise, neck schools or single sessions of extension retraction exercises. Clinicians are encouraged to incorporate these findings into their practice when planning the management of mechanical neck disorders. There is great need for well-designed RCTs to further investigate the topic and perhaps evaluate exercise effectiveness in relation to more specific disorders, e.g., discogenic vs facet joint originated disorder. PMID- 12586558 TI - Neuromuscular control of walking with chronic low-back pain. AB - The reported association of low-back pain and musculoskeletal disorders contributed to the examination of the lumbar spine and hip extensor activation patterns in back pain sufferers during walking. Seventeen idiopathic low-back pain male subjects and 16 healthy volunteers participated in the study. Hip joint ROMs in the sagittal plane and neuromuscular activities of erector spinae [L3, T12], gluteus maximus and biceps femoris were recorded on one randomly selected body side in each group. Analysis using the Student's t-test revealed significant differences for hip joint range of motion, stride time and significantly earlier onsets of the lumbar spine and hip extensors of the back pain sufferers compared with the healthy controls. It is assumed, that low-back disorders are related to changes of the lumbar spine and hip extensor recruitment pattern. PMID- 12586559 TI - Psychometric properties of the Tampa Scale for kinesiophobia and the fear avoidance beliefs questionnaire in acute low back pain. AB - The transition from acute to chronic low back pain (LBP) is influenced by many interacting factors. Pain-related fear, as measured by the Tampa Scale for Kinesiophobia (TSK) and the Fear-Avoidance Beliefs Questionnaire (FABQ), is one of these factors. The objectives of this study were to investigate, in a population with acute LBP, the reliability of TSK and FABQ through evaluation of the internal consistency, the test-retest reliability, and the concurrent validity between TSK and FABQ. One hundred and Seventy-Six patients suffering LBP for no longer than 4 weeks completed a Visual Analogue Scale for pain (VAS), the TSK, the FABQ, and a socio-demographic questionnaire. Each patient completed the VAS, TSK, and FABQ twice within 24 h. Internal consistency of TSK and FABQ scores range from alpha=0.70 to 0.83. Test-retest reliability ranges from r(s)=0.64 to 0.80 (P<0.01). Concurrent validity is moderate, ranging from r(s) =0.33 to 0.59 (P<0.01). It may be concluded that in a population with acute LBP, both the TSK and the FABQ are reliable measures of pain-related fear. In the clinical setting they may provide the practitioner a means of identifying pain-related fear in a patient with acute LBP. PMID- 12586560 TI - Does tape facilitate or inhibit the lower fibres of trapezius? AB - The application of tape to the skin overlying the lower fibres of trapezius is generally thought to facilitate this muscle. However, this facilitation has not been thoroughly investigated. In this study, the effect of tape upon trapezius motoneurone pool excitability was assessed using the trapezius H reflex. The amplitude of the H reflex was measured across four conditions: before tape application, with Endura Fix tape, with the addition of Endura Sports tape and finally with the tape removed. Instead of the expected facilitation of lower trapezius, this tape inhibited lower trapezius activity. On average, the application of Endura Fix tape inhibited trapezius by 4%. The application of Endura Sports tape overlaying the Endura Fix tape inhibited trapezius on average by 22%. This inhibition did not last once the tape was removed. This suggests that any change in shoulder girdle symptoms or movement, which occurs with the application of this particular tape, is not explicable on the basis of the facilitation of the lower fibres of trapezius. PMID- 12586565 TI - Multidisciplinary antimicrobial management and the role of the infectious diseases pharmacist--a UK perspective. AB - Improved clinical outcome, patient safety, cost savings and a reduction in the burden of antimicrobial resistance are outcomes associated with optimizing antimicrobial use. Despite this, the misuse of antimicrobials in the hospital setting remains a huge problem. The development of antimicrobial management teams and the promotion of the role of the clinical pharmacist in antimicrobial prescribing are recommended strategies for improving prescribing practice. It is recognized that there is a lack of published evidence-based research looking at the effects of antimicrobial control programmes and there is a need for more data. In the UK, the role of the hospital pharmacist in promoting responsible antimicrobial prescribing has been largely undervalued and needs to be encouraged and formalized in line with current directives. Managerial structures within hospitals need to endorse multidisciplinary antimicrobial management schemes with appropriate authoritative, administrative and information technology support. PMID- 12586566 TI - Principles of good use of antibiotics in hospitals. AB - Rational use of antimicrobials is a key element for a successful strategy against development of resistance to antimicrobials. The physician should establish the need and the reason for therapy, select the appropriate antimicrobial agent, and then decide on the optimum dose and dosing interval, duration, as well as route of administration. In a particular hospital, the present status of antimicrobial use should be determined, and a strategy should be developed to improve it. This usually encompasses a combination of educative, facilitative and restrictive measures. Good infection control practice is a critical component for success of such a programme. PMID- 12586567 TI - Healthcare-associated outbreak due to pan-drug resistant Acinetobacter baumannii in a surgical intensive care unit. AB - Acinetobacter baumannii is ubiquitous and has recently become one of the most important healthcare-associated (HA) pathogens in hospitals. Infection caused by this organism often leads to significant morbidity and mortality. Outbreaks of pan-drug resistant Acinetobacter baumannii (PDRAB) have rarely been reported. During a two-month period, an outbreak of PDRAB colonization and infection affecting 7 patients occurred in our surgical intensive care unit (SICU). The colonized sites were respiratory tract (N = 7) and central venous catheter (N = 2). One of the patients had a surgical wound infection. Extensive environmental contamination was identified, including sites such as bed rails, bedside tables, surface of ventilators and infusion pump, water for nasogastric feeding and ventilator rinsing and sinks. All of the isolates were analysed by pulsed-field gel electrophoresis (PFGE) and showed an identical pattern. After use of strict cohort nursing, hand hygiene environmental cleaning, and replacement of a dysfunctional high-efficiency particulate air filter (HEPA), the outbreak was controlled. PMID- 12586568 TI - Clonal spread of methicillin-resistant Staphylococcus aureus in a large geographic area of the United States. AB - Methicillin resistance in Staphylococcus aureus has rapidly increased over the last two decades. This increase is paralleled by the emergence of unique multi resistant MRSA clones. In Brazil, Argentina, Uruguay, Portugal and Czech Republic a specific MRSA clone is widely spread, the so-called Brazilian epidemic clone. Another epidemic clone, the Iberian clone, is disseminated in Spain, Portugal, Belgium, Scotland, Italy, Germany and New York. Thus, a large number of hospital acquired infections have been caused by specific MRSA clones. Using different molecular techniques for MRSA typing, we verified that two unique epidemic clones are spread over large geographic area in the US. In addition, we showed that a previously described MRSA clone type, the New York clone (I::A:A), is widely spread beyond the New York frontiers. PMID- 12586569 TI - Nosocomial infections after cardiac surgery in infants and children: incidence and risk factors. AB - This prospective study was undertaken to determine the spectrum, sites and main risk factors for hospital-acquired infections (HAI) in our paediatric cardiothoracic intensive care unit (PCICU), and to determine the main organisms causing bloodstream infection in this setting. All patients admitted between January and December 1999 were prospectively followed for the development of HAI. To define risk factors, patients were grouped by age, complexity score, length of stay in PCICU, and whether the patient's chest was open or closed postoperatively. Three hundred and thirty-five patients underwent cardiac surgery. Fifty-five patients acquired 69 HAIs (HAI patient rate 16.4%). The most common HAI were bloodstream and surgical wound infection in 10 and 8%, respectively. The main causative organisms were Klebsiella spp.,Enterobacter spp. and Pseudomonas spp. in 22, 17 and 16% of episodes, respectively. Staphylococcus spp. accounted for 16% of episodes. The main risk factors for developing HAI were: neonatal age [P < 0.05, odds ratio (OR): 5.89, 95% confidence interval (CI): 2.96-11.58] prolonged PCICU stay (P < 0.05, OR: 6.82, 95% CI: 3.37-14.48), open chest postoperatively (P < 0.05, OR: 3.44, 95% CI: 1.31-8.52) and high complexity score (P < 0.05, OR: 4.03 95% CI: 1.87-8.43). The main causative organisms of bloodstream infections in children hospitalized in the PCICU differ from those in adult and pediatric general intensive care units (ICUs) and include mainly Gram-negative bacilli. High complexity score, neonatal age, prolonged ICU stay, and open chest postoperatively are risk factors of HAI in this patient population. PMID- 12586570 TI - Varicella and paediatric staff: current practice and vaccine cost-effectiveness. AB - This study was undertaken to audit staff varicella policy for UK paediatric centres, and to estimate the cost-effectiveness of implementing a staff varicella vaccine policy. A telephone survey of 22 hospitals was performed to determine the policy regarding varicella immune status among healthcare workers (HCWs). All hospitals surveyed except one recorded immune status of informed HCWs, and had a policy of exclusion from work if they developed chickenpox. The total cost of vaccine implementation for nurses at the John Radcliffe Hospital neonatal unit over 5 years was estimated at 1601 pounds sterling versus 2474 pounds sterling for the estimated total cost of varicella exposure. Therefore it is likely that vaccination represents a cost-effective intervention. PMID- 12586571 TI - Transmission of HCV infection among long-term hospitalized onco-haematological patients. AB - Hepatitis C virus (HCV) infection is becoming a substantial problem in long-term hospitalized patients. Onco-haematological patients undergoing chemotherapy are especially prone to HCV infection. These patients are usually immunosuppressed and therefore antibodies to HCV are not produced despite the presence of HCV RNA in peripheral blood. The aim of the study was to see how often long-term hospitalized patients acquired HCV infection, and what were the possible sources and routes of virus transmission. The study involved 129 children with lymphoproliferative diseases, 36 patients with solid tumours, and 61 healthcare workers from onco-haematological wards. All were HCV RNA and anti-HCV negative at the time of first hospitalization. During a two and a half-year follow-up study among 165 onco-haematological patients, HCV RNA appeared in 87 in subsequent hospitalizations. The majority of infections were (82/87) were 1a genotype, 2 were 1b, 1 was 1a + 1b and 1 was 1a + 3a. In an attempt to establish the origin of HCV infection, healthcare workers were screened for HCV genotyping. All HCV infected staff working on wards had the same genotype (1a). None of the staff was infected with 1b genotype. As the most prevalent genotype in Polish blood donors is 1b, HCV infection in onco-haematological patients is most likely due to horizontal transmission, probably involving genotype 1a, and potential horizontal transmission of HCV is implied by the presence of 1a genotype of HCV in saliva and urine of selected patients. Spread of hospital HCV infection among children may be facilitated by micro-injury of the skin and mucosa. Early detection of HCV RNA is important in such immunosuppressed patients, as they are not able to produce anti-HCV antibodies. This may enable the introduction of prophylactic steps to prevent the spread of HCV infection by horizontal transmission. PMID- 12586572 TI - Comparison of the disinfectant efficacy of Perasafe and 2% glutaraldehyde in in vitro tests. AB - Two percent glutaraldehyde has been the reference disinfectant for high-level disinfection, but its frequent association with adverse effects has stimulated a search for newer disinfectants. We compared the efficacy of 2% glutaraldehyde with that of a new disinfectant formulation, Perasafe (0.2% peracetic acid) in six in vitro tests: germicidal effect in a suspension of micro-organisms; using a microbe-contaminated metallic endodoncy file; an endoscope model; a modified capacity test; a sporicidal test, and corrosiveness on metallic instruments. Both products were effective germicides in 10-20 min, completely destroying microbial inocula except for Mycobacterium and spores. Internal irrigation of endoscopes with 100 mL of either was totally effective. Both substances resisted inactivation after repeated inoculation and did not corrode clean instruments; however, when organic matter was added the 0.2% peracetic acid formulation cleaned without corrosion, while 2% glutaraldehyde fixed the matter to the scalpel, causing corrosion within 2 h. In summary, Perasafe is a good substitute for 2% glutaraldehyde for high-level disinfection. PMID- 12586573 TI - Pharmacokinetics of the antimicrobial agents rifampicin and miconazole released from a loaded central venous catheter. AB - The time course of rifampicin and miconazole concentrations after insertion of a polyurethane catheter loaded with these antibiotics were studied. Data from controlled release experiments in vitro were used, and the concentration time courses of the antimicrobials in serum were calculated by pharmacokinetic simulations. Systemic therapy using typical dosages (rifampicin 600 mg/day iv, miconazole 3 x 200 mg/day iv) results in rifampicin concentrations between 54 and 8424 microg/L, and miconazole concentrations between 3567 and 4676 microg/L. After insertion of a polyurethane catheter loaded with these antibiotics, the maximal concentrations after catheter placement were determined as 6 microg/L at 10.7h for rifampicin, and 13 microg/L at 28.6 h for miconazole. Assuming that the total amount of antibiotics incorporated in the catheter matrix were bioavailable ('worst case'), the resulting maximal concentrations calculated by simulation are 10 microg/L for rifampicin and 65 microg/L for miconazole. Maximal concentrations of rifampicin or miconazole resulting from the insertion of a polyurethane catheter loaded with these antibiotics are, therefore, far below the concentrations resulting from a systemic therapy with the same antimicrobial agents. Even in the worst case, the danger of selecting resistant bacterial strains seems remote because the systemic drug levels are magnitudes of order below subinhibitory concentrations. PMID- 12586574 TI - Bacterial contamination of surgical gloves by water droplets spilt after scrubbing. AB - Wound infection and deep sepsis can have disastrous consequences, particularly in orthopaedic surgery. Strict protocols, ultra-clean air, prophylactic antibiotics, and impervious gowns and drapes, have all been shown to diminish wound infection. However it remains a common and significant problem. The water droplets spilt from the surgeons hands after meticulous scrubbing with povidone iodine were cultured. The permeability of the surgical glove packaging to Gram-positive bacteria was also investigated. The water droplets from the surgeon's arms contained environmental and potentially pathogenic bacteria including a micrococcus, a coliform and coagulase-negative staphylococci. The paper packaging for the range of sterile surgical gloves tested was discovered to be permeable to Gram-positive bacteria. In conclusion accidental water droplet contamination of surgical gloves is a potential source of infection. Alternative recommendations are made. PMID- 12586575 TI - Modified syringe design prevents plunger-related contamination--results of contamination and flow-rate tests. AB - The usual mechanism for syringe contamination is spread of micro-organisms along the inside of the barrel. Two or more full strokes of the plunger will inevitably transport a contaminant from the inside wall into the sterile compartment. We modified syringes by replacing the plunger with a modified version that has an additional central O-ring seal. Conventional and modified syringes were tested for their susceptibility to contamination using standard microbiological and radioisotope methods, and the flow-rate stability of the modified syringe design was assessed in a model with commercially available infusion pumps. Although conventional syringes consistently became contaminated with the second full excursion of the plunger, no contamination was observed in modified syringes, even after 100 aspiration/injection cycles. With respect to flow-rate stability, the new syringe design complies with DIN (German Institute for Standardization) standards. PMID- 12586576 TI - Timing and aetiology of bacterial infections in a liver intensive care unit. AB - We undertook a prospective study of 887 consecutive adult patients admitted over an 11 year period to a liver intensive care unit. One or more bacterial infections occurred in 335 (37.8%) patients. Gram-positive cocci predominated. In relation to the date of admission these infections occurred in a statistically significant sequence. Streptococci infections were earliest (median time to infection two days), followed by Staphylococcus aureus (three days), coagulase negative staphylococci (six days) and enterococci (eight days). Escherichia coli infections occurred earlier than those due to klebsiella-enterobacter (two vs seven days; P = 0.0001) and, overall, Enterobacteriaceae earlier than non fermentative Gram-negatives (four vs. eight days; P = 0.0081). This study contributes to the management of high-dependency patients by confirming statistically the timing and sequence of infecting bacteria in patients with acute liver failure. PMID- 12586577 TI - Should we routinely disinfect floors? PMID- 12586578 TI - Dry cleaning or wet mopping: comparison of bacterial colony counts in the hospital environment. PMID- 12586579 TI - Ultraclean ventilation in operating theatres--beyond laminar flow. PMID- 12586580 TI - Higher disinfectant resistance of nosocomial isolates of Klebsiella oxytoca: indicator organisms in disinfectant testing are not reliable. PMID- 12586581 TI - Sterilization issues in vCJD: significance for prophylaxis. PMID- 12586582 TI - Spread of a dominant methicillin-resistant Staphylococcus aureus (MRSA) clone between Uruguayan and South of Brazil Hospitals. PMID- 12586583 TI - 'I know an old lady who swallowed a fly': a case of (hospital-acquired) human intestinal myiasis. PMID- 12586584 TI - Still learning how to listen to patients. PMID- 12586585 TI - Women's perceptions about treatment decision making for ovarian cancer. AB - OBJECTIVES: To identify in women with advanced epithelial ovarian cancer who had just undergone surgery the extent to which they (1) perceived that they had treatment options, (2) understood the treatment related risks and benefits, and (3) preferred to participate in the treatment decision-making process. METHODS: This qualitative study included women who underwent initial surgery for stage 3 or 4 ovarian cancer and who had received less than two cycles of chemotherapy. In depth semistructured interviews were conducted with 21 patients between June 1999 and February 2001. The interviews were content analyzed according to the themes that arose in the interview. RESULTS: Five themes were identified. (1) Knowledge of treatment benefits and risks. Women understood that the treatment had both survival and quality of life benefits. Women could clearly articulate the risks of chemotherapy. (2) Readiness to make a decision. When making treatment decisions, women described being overwhelmed by the effects of concurrent drugs like analgesics, the severity of the illness, unexpected diagnosis of cancer and grief, and feeling pressured into a decision. (3) Perception of a treatment choice. Most women felt that they made their treatment decision; however, most women did not perceive that they had a treatment choice. Thus, treatment decision making is really a process of coming to terms with the disease and the recommended treatment. (4) Physician-patient relationship. All women suggest that their doctor knew the right treatment for them and they felt confident in their cancer physician. (5) Social supports. Women described supports through decision making processes that included individuals who advocated for them, faith, and past experience with the cancer system. Hindrances to decision making included people who were negative, the cancer label, and employers. CONCLUSIONS: Women with advanced epithelial ovarian cancer did not describe the treatment decision making process as shared; rather they described an interaction that was directed largely by the physician. These women attribute this form of decision-making to their advanced age, severity of illness, immediate ramification of treatment choices, and lack of advocacy for a different model of interaction. Thus, the onus is on the physician to ensure that there is an environment for shared decision-making in the event that the patient is interested in such an interaction. PMID- 12586586 TI - The economic burden of gynecologic cancers in California, 1998. AB - OBJECTIVE: Our aim was to estimate the direct and indirect costs of gynecologic cancers including cervical, ovarian, and uterine cancer in California in 1998. METHODS: Hospitalization costs, including costs of primary and secondary diagnoses of each of the gynecologic cancers, are derived from the California Patient Discharge dataset. Charges are converted to costs using hospital-specific cost-to-charge ratios and an imputed cost for HMO hospitalizations. Other direct medical costs are derived from the 1997 Medical Expenditure Panel Survey. Indirect mortality costs are the product of the number of deaths and the expected value of a female's future earnings, including an imputed value of housekeeping services. RESULTS: The total cost of the three gynecologic cancers in California is estimated at $624 million for 1998. Indirect costs or losses in productivity due to premature death are twice the direct medical care costs. Hospitalization inpatient costs account for more than half of total direct costs for each of these cancers. Total costs for ovarian cancer are highest among the three cancers. CONCLUSIONS: The estimates presented here highlight the need for studies that identify cost-effective screening methods and the relationship between treatment protocols and outcomes. The methods presented here could be used to develop stage- or treatment-specific costs that would be useful for such analyses. Only through early detection and more effective management of gynecologic cancers can there be a reduction in the morbidity, premature death, and high costs of cervical, ovarian, and uterine cancers. PMID- 12586587 TI - Risk of abnormal pregnancy completing chemotherapy for gestational trophoblastic tumor. AB - OBJECTIVE: This study analyzed the outcome of the first pregnancy following chemotherapy for gestational trophoblastic tumor (GTT). METHODS: A total of 387 patients with GTT (85 patients with high-risk GTT and 302 patients with low-risk GTT) underwent chemotherapy at Chiba University Hospital between 1974 and 2000. Of these patients, 130 women (18 with high-risk GTT and 112 with low-risk GTT), who achieved remission and had at least one conception following chemotherapy, were included in the study. RESULTS: The outcomes of all the first subsequent pregnancies in women treated with methotrexate, actinomycin-D, or etoposide (including those switched to other regimens), or combination therapy, were comparable to those in the Japanese general population. However, the incidence of abnormal pregnancies (spontaneous abortion, still birth, repeat mole) was significantly higher in women who conceived within 6 months of completing chemotherapy (4/15; 40%) than in those who conceived after the recommended waiting period of more than 12 months (10/95; 10.5%) (P = 0.028). CONCLUSION: Patients with GTT who achieved remission after chemotherapy with methotrexate, actinomycin-D, or etoposide, or combination therapy, may anticipate a normal future reproductive outcome. As pregnancies occurring within 6 months following remission are at risk of abnormalities, a waiting period of at least 6 months after chemotherapy for GTT is suggested. PMID- 12586588 TI - Mechanisms involved in the evolution of progestin resistance in human endometrial hyperplasia--precursor of endometrial cancer. AB - BACKGROUND: Successful treatment of endometrial hyperplasia with progestins is commonly accompanied by the finding of an inactive or suppressed endometrium after therapy. However, approximately 30% of the endometrial hyperplasia cases do not respond to progestins and hyperplastic glands persist. The Fas/FasL system is known to play a role in tissue remodeling as a result of changes in menstrual hormone levels. The aims of this study are to examine Fas/FasL expression in endometrial hyperplasia of pre- and postprogestin treatment samples and to study the Fas/FasL regulation in vitro with Ishikawa cells after progestin stimulation. DESIGN: Pre- and posttreatment paraffin-embedded endometrial hyperplasia tissue samples from 26 women were examined by immunohistochemistry for changes in Fas/FasL expression related to the administration of progestins. Among 26 patients, 18 were successfully treated with progestins and 8 failed treatment. Fas/ FasL positivity was defined by the presence of 10% or more immunoreactive epithelial cells in each specimen. In positive cases, a percentage or an immunoscore of immunoreactive cells was given by counting 500 cells. Cell viability was evaluated by the MTT assay. The in vitro effects of progesterone on Fas/FasL expression and apoptosis in Ishikawa cells were examined by using Western blot and TUNEL assays, respectively. RESULTS: Fas immunoreactivity was present in 4/26 (15%) preprogestin cases with an average of 16% of the epithelial cells expressing Fas. FasL was expressed in 21/26 (80%) pretreatment cases with an average of 42% of the hyperplastic glandular cells being positive. In postprogestin cases, an increase of Fas expression (14/18, 77%) with an average of 47% stained cells was seen in responders (P < 0.001), while FasL was found in 16/18 (89%) responders with an average of 65% of cells positive (P = 0.587). In nonresponders, no significant changes in Fas/FasL expression were detected compared to pretreatment samples. With in vitro Ishikawa cells, a slight increase (10-20%) of Fas and FasL protein expression was detected after 24 h of progesterone treatment, but a more significant increase (220-343%) of both Fas and FasL expression was found after 48 h of withdrawing progesterone, which parallels apoptotic activity. CONCLUSIONS: The Fas/FasL system may be involved in the development of endometrial hyperplasia. Part of the molecular mechanisms of progestin therapy for endometrial hyperplasia is through upregulation of Fas/FasL expression. Dysregulation of Fas/FasL expression in hyperplastic endometrium may be part of the molecular mechanisms for nonresponders to progestin treatment. Intermittent, rather than continuous, progestin treatment may be more effective clinically for the treatment of endometrial hyperplasia. PMID- 12586589 TI - Altretamine (hexamethylmelamine) in the treatment of platinum-resistant ovarian cancer: a phase II study. AB - OBJECTIVE: To evaluate the activity of oral Altretamine in women with epithelial ovarian carcinoma who responded (PR or CR) to first line chemotherapy but relapsed within 6 months. The protocol was later amended to include patients with relapse within 12 months. METHODS: A multicentric phase II trial. The patients had to have measurable disease. No more than one prior chemotherapy regiment was allowed. The patients were treated with 260 mg/m(2)/day of Altretamine in four divided doses for 2 weeks, repeated every 4 weeks. The response was evaluated after every two courses. RESULTS: Thirty-one eligible patients were treated with a median of 3 courses of Altretamine (range 1-12). Hematological toxicity was minimal. Gastrointestinal toxicity was common. Response evaluation was possible for 26 patients. Three patients (9.7% intent-to-treat) achieved a partial response. Eight patients had stable disease, and 15 patients had progressive disease after two treatment courses. The median time to progression was 10 weeks (range, 5-51 weeks). Medial survival was 34 weeks (range, 7-112+). CONCLUSION: Altretamine should not be chosen as standard treatment in patients with platinum resistant recurrent ovarian cancer. However, Altretamine represents a useful alternative in patients who prefer oral treatment or when socioeconomic considerations are an important issue. PMID- 12586590 TI - Overexpression of the c-Met/HGF receptor and its prognostic significance in uterine cervix carcinomas. AB - OBJECTIVE: The purpose of this study is to evaluate the significance of the c Met/hepatocyte growth factor receptor expression in invasive cervical carcinoma. METHODS: Ninety-Four patients with FIGO stage 1B disease, treated primarily with surgery, were studied immunohistochemically. Of the cases, 67 were squamous carcinoma and 27 were nonsquamous (10 were adenocarcinoma, 15 were adenosquamous carcinoma, and 2 were indifferentiated carcinoma). Immunohistochemically stained c-Met slides of primary malignancies were evaluated blindly of clinical outcome and other histopathological factors. RESULTS: Overexpression of c-Met was found in 56 of 94 specimens. Primary tumors which show recurrences were found to be c Met overexpressors. Univariate survival analysis (Kaplan-Meier) showed that c-Met overexpression is significantly correlated with disease-free survival. Moreover the diameter of the primary tumor, deep cervical stromal invasion, presence of metastatic lymph node, number of metastatic lymph nodes and c-Met overexpression were significantly correlated with overall 5-year survival. Furthermore multivariant analysis with Cox regression showed that the presence of metastatic lymph node and immunopositivity for c-Met are significantly correlated with overall survival, while c-Met overexpression was found to be an independent variable for disease-free survival. CONCLUSION: These results reveal that c-Met oncogene overexpression is an important parameter for disease progression, recurrence, and survival in early-stage invasive uterine cervix carcinomas. PMID- 12586591 TI - A phase II study of docetaxel in paclitaxel-resistant ovarian and peritoneal carcinoma: a Gynecologic Oncology Group study. AB - OBJECTIVES: Docetaxel is an inhibitor of microtubule depolymerization and has demonstrated activity in paclitaxel-resistant breast cancer and gynecologic cancer. The Gynecologic Oncology Group (GOG) conducted a study of docetaxel in paclitaxel-resistant ovarian and peritoneal carcinoma to determine its activity, and nature and degree of toxicity, in this cohort of patients. METHODS: Patients with platinum- and paclitaxel-resistant ovarian or peritoneal carcinoma, defined as progression while on or within 6 months of therapy, were eligible if they had measurable disease and had not received more than one chemotherapy regimen. Docetaxel at a dose of 100 mg/m(2) was administered iv over 1 h every 21 days. A prophylactic regimen of oral dexamethasone 8 mg bid was begun 24 h before docetaxel administration and continued for 48 h thereafter. Hepatic function was strictly monitored. RESULTS: Sixty patients were entered and treated with a total of 256 courses, with all 60 evaluable for toxicity and 58 evaluable for response. Responses were observed in 22.4% of patients, with 5.2% achieving complete response and 17.2% achieving partial response (95% CI, 12.5-35.3%). The median duration of response was 2.5 months. The likelihood of observing a response did not appear to be related to the length of the prior paclitaxel-free interval or duration of prior paclitaxel infusions. The principal adverse effect of grade 4 neutropenia occurred in 75% of patients. There was one treatment-related death. Dose reductions were required in 36% of patients. CONCLUSIONS: Docetaxel is active in paclitaxel-resistant ovarian and peritoneal cancer but, in view of significant hematologic toxicity, further study is warranted to ascertain its optimal dose and schedule. PMID- 12586592 TI - Three-consecutive-day topotecan is an active regimen for recurrent epithelial ovarian cancer. AB - OBJECTIVE: The aim was to determine the response rate and toxicity of topotecan administered Days 1-3 every 21 days for recurrent epithelial cancers of the ovary, peritoneum, or fallopian tube. A 3-day regimen may be more convenient and less expensive than a 5-day schedule. METHODS: Patients with recurrent epithelial cancer of the ovary, peritoneum, or fallopian tube who had adequate hepatic, renal, and hematologic function were eligible for participation. Topotecan (2 mg/m(2)) was administered for 3 consecutive days every 21 days. Response was measured clinically and serologically. Granulocyte colony stimulating factors (GCSF) were not utilized prophylactically, but could be added under specific conditions. RESULTS: Thirty-one patients with recurrent ovarian cancer whose median age was 63 (range 32-84) received 165 cycles of topotecan (median = 6; range 2-8) and are evaluable for toxicity. The median number of prior regimens was 1. Topotecan was administered on schedule in 96.6% of cycles. Grade 3/4 neutropenia was seen in 29.1 and 23.6% of courses, respectively; but only 3.4% of cycles required GCSF support (6 cycles for 2 patients). Grade 4 thrombocytopenia was rare (1% of cycles). Nonhematologic toxicity was mild. The response rate for 28 evaluable patients was 32.1% (10.7% complete response (CR) and 21.4% partial response (PR)); stable disease was seen in 17.9% of patients. The median progression-free interval (PFI) for all patients was 15.5 weeks (range 5-40). Eighteen platinum-sensitive patients demonstrated a 43.4% response rate (12.5% CR and 31.3% PR); stable disease was documented in 18.8%. The median PFI for platinum-sensitive patients was 18.5 weeks (range 5-40). CONCLUSION: Topotecan is an effective regimen with acceptable toxicity for recurrent ovarian cancer when administered for 3 consecutive days (2 mg/m(2)) every 21 days. It can be delivered on schedule without GCSF support in the vast majority of patients. PMID- 12586593 TI - Ovarian cancer in Western Australia (1982-1998): trends in surgical intervention and relative survival. AB - OBJECTIVES: We aimed to review the utilisation and trends in surgical procedures for the primary management of ovarian cancer and the survival outcomes of patients surgically treated in Western Australia. METHODS: The population-based Western Australia Data Linkage System was used to link hospital morbidity and mortality data for all women diagnosed with malignant primary ovarian cancer in the State Cancer Registry in the period 1982-1998. Poisson regression was used to analyse trends in surgical procedure rates. Logistic regression examined the likelihood of having a surgical procedure in the periods 1988-1993 and 1994-1998 compared with 1982-1987. Relative survival was used to adjust survival estimates for other causes of death occurring in the general female population. RESULTS: There were 1,126 women who underwent a primary surgical procedure for ovarian cancer in Western Australia in the period 1982-1998. Women were more likely to undergo surgery in 1994-1998 (87.8%) compared with 1988-1993 (76.8%), but there was no difference when compared to 1982-1987 (89.2%) (P = 0.62). The likelihood of using specific surgical procedures to treat ovarian cancer increased for all but total abdominal hysterectomy. Bilateral salpingo-oophorectomy was 3.7 times more likely to be performed and omentectomy 5 times more likely to be performed in 1994-1998 compared with 1982-1987. The median length of hospital stay decreased from 15 to 12 days and emergency admissions decreased from 26.5 to 15.4% over the three time periods. Thirty-two percent of women were readmitted within 30 days of separation from their primary surgery, 23% of which were for the same-day treatment with either chemotherapy or radiotherapy. A 15% increase in relative survival was observed between the periods 1982-1997 (38.8%) and 1994 1998 (53.5%).Conclusion. CONCLUSION: Surgery remains a cornerstone in the primary management of ovarian cancer. There have been dramatic shifts in surgical practice in Western Australia, with more women undergoing certain surgical procedures today than they were 20 years ago. Coupling the increasing surgical trends are improved outcomes. Fewer women are presenting as an emergency, the length of hospital stay has been reduced, and survival outcomes have shown a significant improvement. PMID- 12586594 TI - Secreting ovarian tumors may protect women from osteoporosis. AB - OBJECTIVE: It is well known that ovarian steroids modulate bone turnover. Conditions associated with low levels of these hormones, such as menopause, hypogonadism, and others, have been related to osteopenia or osteoporosis. On the other hand, hyperandrogenism in premenopausal women, mainly in polycystic ovarian syndrome, has been reported to have a protective effect on bone mass. However, data regarding how bone mass is affected by neoformative processes in which steroids are increased are not as well documented. Our aim was to study the effect of secreting ovarian tumors on bone mass. METHODS: A total of 14 patients were referred to our hospital because of endocrine ovarian tumors. Steroid levels were measured prior to and after surgery. Bone mineral density (BMD) by DEXA was assessed at inclusion in all cases. Additionally, in 7 women bone measurement was repeated after 1-year follow-up. The setting was a tertiary hospital. RESULTS: All patients showed increased levels of testosterone, androstenedione, and free testosterone prior to surgery. BMD was also in the normal-upper range or over normal in all of them. As expected in the subjects with a second DEXA a decrease in bone mass was noted. CONCLUSION: Steroid secreting ovarian tumors increase bone mass and thus may protect women from later osteoporosis. PMID- 12586595 TI - Landmark contribution of Hans-Ludvig Kottmeier to management of early-stage endometrial cancer. PMID- 12586596 TI - Antimetastatic and immunomodulating properties of a new herbal prescription, Bojung-bangam-tang. AB - We investigated the antimetastatic effect of Bojung-bangam-tang, a new herbal prescription, on liver metastasis by the inoculation of colon 26-L5 carcinoma cells into the portal vein. Oral administration of Bojung-bangam-tang for 15 days from day 7 before tumor inoculation significantly inhibited liver metastasis in a dose-dependent manner. Bojung-bangam-tang enhanced the mitogenic activity of BALB/c whole splenocytes induced by various mitogenic stimuli. Oral administration of Bojung-bangam-tang, by itself, could not induce the production of interleukin (IL)-12 or IFN-gamma by macrophages, but enhanced the potential of macrophages to produce cytokines in response to lipopolysaccharide (LPS). Experiments using macrophage- or natural killer (NK) cell-deficient mice revealed that the antimetastatic effect of Bojung-bangam-tang is mediated by macrophages rather than NK cells. Bojung-bangam-tang caused a marked increase of production of Th1 cytokine (IFN-gamma) and decrease of production of Th2 cytokine (IL-4) by splenocytes stimulated with concanavalin A (Con A). These results indicated that oral administration of Bojung-bangam-tang inhibited the liver metastasis of colon 26-L5 cells through a mechanism leading to a Th1 dominant immune state and activation of macrophages. Thus, Bojung-bangam-tang may be useful for the prevention of cancer metastasis. PMID- 12586597 TI - A novel prodigiosin-like immunosuppressant from an alkalophilic Micrococcus sp. AB - A novel red pigment, 2,2'-[3-methoxy-1'amyl-5'-methyl-4-(1-pyrryl)] dipyrryl methene (MAMPDM), which has properties similar to those of prodigiosins, has been isolated for the first time from a bacterium putatively identified as Micrococcus sp. Our studies showed that MAMPDM inhibited proliferation of both human T as well as B cells and murine T cells, in response to polyclonal mitogens, in a concentration-dependent manner while murine B cell proliferation induced by lipopolysaccharide was inhibited only at high concentration. The effect of MAMPDM on constitutive cell cycling was ascertained using four mouse and human tumour cell lines. At 100 nM, the concentration that inhibited con A induced proliferation of mouse spleen cells, the viability of these cell lines was not affected. At 10-100-fold higher concentration of MAMPDM, however, there was a decrease in cell viability with T cell-derived cell lines being more sensitive. MAMPDM did not block the secretion of IL-2 or expression of CD25 though it inhibited the proliferation of con A stimulated T cells. The higher amount of IL 2 in the supernatant of the con A stimulated T cells, cultured in the presence of the immunomodulator, indicated accumulation of IL-2 due to its reduced utilisation. At inhibitory concentration, MAMPDM induced apoptosis in con A stimulated cells. Thus, MAMPDM may have considerable and selective T cell immunosuppressive potential and appears to act by a mechanism distinct from that of other known immunosuppressors. PMID- 12586598 TI - Prednisolone inhibits LPS-induced bone marrow suppressor cell activity in vitro but not in vivo. AB - Glucocorticoids are used clinically to treat a variety of inflammatory diseases including endotoxemia. We hypothesized that injecting mice with the steroid prednisolone (pred) would mitigate the enhanced bone marrow (BM) natural suppressor (NS) cell activity that occurs in mice after receiving an injection of lipopolysaccharide (LPS). In vitro, prednisolone blocked the ability of NS cells to produce the immunosuppressive molecule nitric oxide (NO) and also the ability to suppress T cell proliferation. Prednisolone acted both indirectly, by blocking synthesis of cytokines necessary for NS cell activation, and also directly on NS cells, by blocking production of NO. In vivo, variable results were obtained. Prednisolone at 20 microg/gm did decrease NS activity when injected into normal mice. However, when mice were injected with both LPS and prednisolone (0.2 or 20 microg/gm), a large increase in BM NS activity was observed. The increase was evident in both the ability of the BM cells to suppress T cell proliferation and to produce NO. The data show that, in vivo, the steroid prednisolone in conjunction with the inflammatory compound LPS act to enhance BM NS activity. PMID- 12586599 TI - Specific inhibitors of protein phosphatase 2A inhibit tumor metastasis through augmentation of natural killer cells. AB - Selective augmentation of natural killer (NK) cells can suppress tumor metastasis, but molecular targets for NK cell activation have not been identified. We report here that cytostatin (CTS), a novel specific inhibitor of protein phosphatase (PP) 2A, can inhibit B16 melanoma pulmonary metastasis by the expansion and activation of NK cells. CTS administration in vivo increased mRNA expression of Flt-3 ligand, one of NK-generating cytokines, in bone marrow cells. Phoslactomycin A and leustroducsin H, other specific inhibitors of PP2A, also augmented NK cell activity and inhibited lung metastasis, but a CTS analogue without inhibitory activity on PP2A and calyculin A, a dual inhibitor of PP1 and PP2A, did not. These results suggest that specific inhibition of PP2A can augment NK cells through upregulation of NK-generating cytokine and prophylaxis for pulmonary metastasis. PMID- 12586600 TI - Effects of anti-lymphocytes and anti-thymocytes globulin on human dendritic cells. AB - Dendritic cells (DC) were implicated as the mediators of primary stimulation of the immune response against donor antigen. Anti-lymphocytes globulin (ALG) and anti-thymocytes globulin (ATG) antibodies have been widely used as an immunosuppressive agent in the treatment and prevention of rejection after organ transplantation. In this study, we attempted to determine whether ATG and ALG are able to bind and interfere in human dendritic cell function. ALG and ATG bound to human DCs at least in part by recognising CD1a, MHC I, MHC II, CD11a, CD86, CD32, CD4, CD11b, CD29 and CD51/61. This binding, that was more relevant in mature DCs, induced complement-mediated lysis. ALG and ATG did not influence either the macropynocytosis of Lucifer Yellow (LY) or the receptor-mediated endocytosis of FITC-Dextran. In MLR assay, ATG and ALG were able to significantly inhibit T-cell proliferation by binding on T cell but not on dendritic cells. CONCLUSIONS: we demonstrate that ALG/ATG is able to interfere in the activation of T cells by dendritic cells in two different ways: by inhibiting the capacity of lymphocyte to proliferate after the DC stimulation and by inducing a complement-mediated lysis of DC. PMID- 12586601 TI - Immunomodulatory activity of TNF-alpha during acute liver injury induced by D galactosamine and its protection by PGE1 in rats. AB - Tumour necrosis factor-alpha (TNF-alpha) mediates hepatocyte cell death by D galactosamine (D-GalN) and its protection by prostaglandin E(1) (PGE(1)). The activation of immune system plays an important role in the development of liver injury. TNF-alpha and PGE(1) regulate the activity and cytokine release of different inflammatory cells. The present study was undertaken to determine if the noxious or hepatic protective properties of TNF-alpha during D-GalN-induced liver injury was related to an alteration by PGE(1) of the immunoregulatory activity of TNF-alpha. The role of TNF-alpha was assessed by anti-TNF-alpha antibodies to D-GalN-treated rats in the presence or absence of PGE(1). D-GalN enhanced the percentage of monocytes and T lymphocytes in the total peripheral blood mononuclear cells (PBMCs). D-GalN enhanced the activation degree of monocytes, but reduced that of T lymphocytes. D-GalN also enhanced TNF-alpha, IL 1alpha, IL-6 and IFN-gamma concentrations in blood. Anti-TNF-alpha antibodies abolished all immunological changes and greatly reduced liver damage induced by D GalN. The protection by PGE(1) against D-GalN liver injury was associated with an increase in TNF-alpha concentration and a reduction of IL-1alpha and IL-6. These changes were associated with a reduction of monocyte activation degree and a recovery of that of T lymphocytes. Although anti-TNF-alpha antibodies abolished the protection by PGE(1) against D-GalN-liver injury, they did not essentially counteract the effect of the prostanoid in all immunological parameters studied. The present study showed that the protection against D-GalN liver damage by PGE(1) or anti-TNF-alpha was associated with similar effects on the inflammatory parameters studied. Nevertheless, the abolishment of liver protection by PGE(1) with anti-TNF-alpha in D-GalN-treated rats in the presence of a protective cytokine profile suggests that the release of TNF-alpha induced by PGE(1) pre administration was exerting a direct protective effect on hepatocytes against D GalN injury. Consequently, the effect of PGE(1) on inflammatory parameters studied during liver injury was unrelated to TNF-alpha. PMID- 12586602 TI - The novel differentiation of human blood mononuclear cells into CD1a-negative dendritic cells is stimulated in the absence of exogenous cytokines by an extract prepared from pinecones. AB - The production of dendritic cells, both in-vivo and in-vitro, has become the intense focus of research activities. Common to many of these production protocols is the use of cytokines, typically granulocyte-monocyte colony stimulating factor and either interleukin 4 or tumor necrosis factor alpha or a combination of all three. Herein, we report our findings that a proprietary pinecone extract is capable of in a dose-dependent manner, and in the absence of exogenous cytokines, the rapid differentiation from peripheral blood mononuclear cells of mature CD1a-negative dendritic cells. PMID- 12586603 TI - Inhibition of TNF-alpha-induced RANTES expression in human hepatocyte-derived cells by fibrates, the hypolipidemic drugs. AB - Increased concentrations and activity of plasma cytokines produced by monocytes, macrophages, and hepatocytes in patients with alcoholic liver diseases, correlate with the clinical course of liver diseases and are of prognostic value. Especially, high levels of circulating tumor necrosis factor (TNF)-alpha have been found to correlate with increased mortality in alcoholic hepatitis. Moreover, hepatic RANTES was increased in patients with alcoholic hepatitis. Thus, TNF-alpha-induced RANTES expression may have a critical role in cell mediated liver injury associated with alcoholic hepatitis. Fibrates are widely used in the treatment of hyperlipidemia and lower triglyceride levels in patients with hyperlipidemia. Recently, several groups reported that bezafibrate, one of fibrates, is effective in primary biliary cirrhosis treatment. Additionally, it is reported that bezafibrate is effective in the treatment not only of primary biliary cirrhosis but also of chronic hepatitis C and tamoxifen-induced non alcoholic steatohepatitis. We, here, presented that bezafibrate and fenofibrate repressed TNF-alpha-induced protein production and mRNA expression of RANTES in human hepatocyte-derived cells. Luciferase assay showed that bezafibrate and fenofibrate inhibited RANTES gene expression in response to TNF-alpha. Moreover, bezafibrate repressed TNF-alpha-induced DNA-binding activity of NF-kappaB. Thus, fibrates reduced TNF-alpha-induced NF-kappaB activation and RANTES expression, possibly suggesting that fibrates might be inhibitory agents of migration of inflammatory cells by RANTES to the liver in patients with alcoholic liver diseases. In line of these results, it might be possible that fibrates are therapeutic agents in alcoholic liver diseases. PMID- 12586604 TI - CD28 co-stimulation regulates the effect of transforming growth factor-beta1 on the proliferation of naive CD4+ T cells. AB - Transforming growth factor-beta1 (TGF-beta1) is a critical regulator of T cell responses in vivo. In vitro, TGF-beta1 can either enhance or inhibit T cell proliferative responses, but the relevant factors that determine the T cell response to TGF-beta1 remain obscure. Here, we present evidence that CD28 co stimulation modifies the effects of TGF-beta1 on T cell proliferation. In the absence of CD28 co-stimulation, TGF-beta1 potently suppressed TCR-stimulated proliferation of naive T cells. In the presence of CD28 co-stimulation, TGF-beta1 potently inhibited T cell apoptosis and enhanced TCR-stimulated proliferation. A similar effect of CD28 co-stimulation was not observed in memory/effector cells, whose proliferation was enhanced by TGF-beta1, whether co-stimulated or not. We examined the mechanism by which CD28 modulates naive T cell responses to TGF beta1. Since CD28 co-stimulation classically is a potent enhancer of interleukin (IL)-2 production, we anticipated observing high IL-2 production from naive T cells stimulated with anti-CD3/anti-CD28 and TGF-beta1. Surprisingly, however, TGF-beta1 strongly inhibited production of IL-2 from naive CD4(+) T cells, even when CD28 was engaged. Even though IL-2 levels were strongly suppressed by TGF beta1 to trace levels, antibody neutralization studies showed that IL-2 is still a basic requirement for the proliferation of anti-CD3/anti-CD28/TGF-beta1 stimulated naive T cells. These data show that CD28's modulation of T cell responses to TGF-beta1 is not via the production of high levels of IL-2, and suggest that engagement of CD28 may activate additional downstream pathways that modulate the responses of naive T cells to TGF-beta1. PMID- 12586605 TI - A Limulus anti-LPS factor-derived peptide modulates cytokine gene expression and promotes resolution of bacterial acute infection in mice. AB - Sepsis in experimental animals and humans has been associated with perturbed immune response. A major event contributing to the decrease in immune functions in septic disorders seems to be the inadequate balance of cytokines mediating the interactions between the innate and adaptive immune systems. We previously observed that a cyclic peptide derived from the Limulus anti-LPS factor (LALF), which partially protect mice from endotoxic shock lethality, has the ability to modulate cytokine secretion in vitro. We herein examined the effects of the LALF(31-52) peptide in an experimental model of Gram-negative peritoneal sepsis and analyzed the cytokine gene expression in the spleen and liver of peptide treated mice. The prophylactic administration of LALF(31-52) abrogated the systemic TNF-alpha response, reduced organ damage and increased the survival of infected mice. Histological examination of spleen and liver in peptide-treated mice showed prevention of tissue damage induced by the high dose of Pseudomonas aeruginosa. This treatment modulates the cytokine gene expression in these tissues, stimulating IL-2, IL-12 and IL-13 mRNA synthesis, while IL-4 and IL-10 mRNA expression was not modified. This cytokine profile induced by the LALF derived peptide seems to be favorable for host resistance against Gram-negative bacteria acute infection. In addition, peptide treatment was effective after the initiation of the systemic inflammatory response, promoting a significant increase in mice survival. These results further demonstrate the immunomodulatory potential of LALF(31-52) and are relevant for the design of prophylactic and therapeutic strategies for acute bacteria infection and sepsis, especially for preventing or ameliorating host immunity defects in these disorders. PMID- 12586606 TI - Immunomodulatory effects of fenugreek (Trigonella foenum graecum L.) extract in mice. AB - Immunomodulatory activity of aqueous extract of Trigonella foenum graecum L., a widely used medicinal and dietary herb, was evaluated in male Swiss albino mice. Mice were treated with three doses of extract (50, 100 and 250 mg/kg body weight per os) for 10 days. Body weight, relative organ weight, cellularity of lymphoid organs, delayed type of hypersensitivity (DTH) response, plaque-forming cell (PFC) assay, haemagglutination titre (HT), quantitative haemolysis of SRBC (QHS) assay, phagocytosis, and lymphoproliferation were studied in various groups of animals. At doses of 50 and 100 mg/kg, a significant increase (p < 0.05) in relative organ weight of thymus was observed but there was no effect on kidney and spleen weights. Liver weight also increased significantly at doses of 100 and 250 mg/kg. However, no elevation in the levels of liver function test (LFT) enzymes was observed. As regards lymphoid organ cellularity, spleen recorded no significant increase at any dose, whereas cellularities of thymus and bone marrow were significantly increased. T. foenum graecum extract elicited a significant (p < 0.001) increase in the DTH response at doses of 50 and 100 mg/kg, but the change at higher dose of 250 mg/kg was not statistically significant. Humoral immunity as measured by PFC showed an elevated response at a dose of 100 mg/kg, but at 50 and 250 mg/kg, no significant effect was observed. In the HT test, plant extract also showed modulatory effect at all the doses. Plant extract elicited a significant increase in phagocytic index and phagocytic capacity of macrophages. Stimulatory response of plant extract was also observed in lymphoproliferation assay but the response was weak. Overall, T. foenum graecum showed a stimulatory effect on immune functions in mice. As it is used for a variety of medicinal purposes, its immunostimulatory effect, as reported in this study, strengthens the rationale of its use in several Ayurvedic and Unani drugs. PMID- 12586607 TI - Immunomodulatory effect of fructose-1,6-bisphosphate on T-lymphocytes. AB - Sepsis remains an important and life-threatening problem, and is the most common cause of death in the intensive care unit. One promising therapeutic candidate for protection against injury in sepsis is fructose-1,6-bisphosphate (FBP), a high-energy glycolytic pathway intermediate. The objective of the study was to establish a role for FBP on the immune system, especially in lymphocyte proliferation. Peripheral blood mononuclear cells (PBMCs) were isolated from the blood of healthy humans by gradient centrifugation. T-lymphocytes were stimulated for 96 h with phytohemagglutinin (PHA) and varying concentration of FBP. Fructose 1,6-bisphosphate at concentrations between 1.2 and 10 mM decreased proliferation of T-lymphocytes and reduced the viability only at concentrations 5.0 and 10 mM. The levels of soluble IL-2 receptor were reduced at FBP concentrations between 1.2 and 10 mM. In conclusion, this study demonstrates that FBP has important effect on immunomodulatory and this result can be correlated with the protection against injury in sepsis. PMID- 12586608 TI - Multiple mechanisms involved in the inhibition of proinflammatory cytokine production from human monocytes by N-(p-coumaroyl)serotonin and its derivatives. AB - We have reported that N-(p-coumaroyl)serotonin(CS) isolated from safflower oil cake (Carthamus tinctorius L.) inhibits the production of proinflammatory cytokines by endotoxin (LPS)- stimulated human monocytes. In this study, the effects of CS and its three derivatives, N-(trans-cinnamoyl)serotonin (Cin.S), N (trans-cinnamoyl)tryptamine (Cin.T), and N-(p-coumaroyl)tryptamine (CT) on the production of proinflammatory cytokines were compared. Cin.S possessed radical scavenging activity at a comparable level to CS, while CT and Cin.T exhibited lower activity, suggesting that hydroxyl group in serotonin is essential for the antioxidative activity. CS and CT strongly inhibited the production of proinflammatory cytokines (IL-1alpha, IL-1beta, IL-6, IL-8, and TNF-alpha) from LPS-stimulated human monocytes. However, Cin.S inhibited the production of only IL-1alpha and IL-1beta, and Cin.T inhibited none of these cytokines production. CS and CT markedly inhibited the protein synthesis in monocytes, the inhibitory effect of Cin.S was moderate, and that of Cin.T was quite weak. These results indicate that CS and its derivatives inhibit the production of proinflammatory cytokines through multiple mechanisms. PMID- 12586609 TI - High-dose cyclophosphamide in multiple sclerosis patients undergoing autologous stem cell transplantation. AB - High-dose cyclophosphamide (CTX) is commonly used in preparation for autologous and allogeneic stem cell transplantation. CTX is a pro-drug, which undergoes complex oxidative metabolism with the metabolites being eliminated both renally and hepatically. In the following study, we evaluated the pharmacokinetic characteristics of high-dose CTX in three patients undergoing autologous stem cell transplantation for multiple sclerosis. The plasma concentration-time profiles for CTX and its hydroxy-metabolite were similar in multiple sclerosis patients to those reported in cancer patients undergoing stem cell transplantation. There was an increase in drug clearance after the second CTX dose indicating that the drug induced its own metabolism consistent with reports in other populations receiving high-dose CTX. One of the three patients cleared the drug slowly but this was not associated with greater toxicity. The patient with the slow clearance value and therefore highest drug exposure had stable disability scores at 2 years posttransplant compared with baseline values taken prior to transplantation. In conclusion, in this small case series, there was no indication that CTX metabolism was different than that in other populations undergoing transplantation. PMID- 12586610 TI - Nucleolar localization of RPS19 protein in normal cells and mislocalization due to mutations in the nucleolar localization signals in 2 Diamond-Blackfan anemia patients: potential insights into pathophysiology. AB - Ribosomal protein S19 (RPS19) is frequently mutated in Diamond-Blackfan anemia (DBA), a rare congenital hypoplastic anemia. Recent studies have shown that RPS19 expression decreases during terminal erythroid differentiation. Currently no information is available on the subcellular localization of normal RPS19 and the potential effects of various RPS19 mutations on cellular localization. In the present study, using wild-type and mutant RPS19 cDNA, we explored the subcellular distribution of normal and mutant proteins in a fibroblast cell line (Cos-7 cells). RPS19 was detected primarily in the nucleus, and more specifically in the nucleoli, where RPS19 colocalized with the nucleolar protein nucleolin. Using various N-terminal and C-terminal deletion constructs, we identified 2 nucleolar localization signals (NoSs) in RPS19: the first comprising amino acids Met1 to Arg16 in the NH2-terminus and the second comprising Gly120 to Asn142 in the COOH terminus. Importantly, 2 mutations identified in DBA patients, Val15Phe and Gly127Gln, each of which localized to 1 of the 2 NoS, failed to localize RPS19 to the nucleolus. In addition to their mislocalization, there was a dramatic decrease in the expression of the 2 mutant proteins compared to the wild type. This decrease in protein expression was specific for the mutant RPS19, since expression of other proteins was normal. The present findings enable us to document the nucleolar localization signals in RPS19 and help define the phenotypic consequences of some mutations in RPS19 in DBA. PMID- 12586611 TI - Activated protein C alters cytosolic calcium flux in human brain endothelium via binding to endothelial protein C receptor and activation of protease activated receptor-1. AB - Activated protein C (APC) exerts endothelial protein C receptor (EPCR)-dependent neuroprotective effects in a brain focal ischemia model and direct cellular effects on human umbilical vein endothelial cells (HUVECs) via protease-activated receptor-1 (PAR-1). Because PAR receptors are expressed in brain endothelium and mediate intracellular calcium concentration ([Ca2+]i) signaling, we hypothesized that APC may regulate intracellular [Ca2+] flux in human brain endothelial cells (BECs) via EPCR and PAR-1. Primary cortical BECs derived from human autopsies (early passage) and HUVECs were used for [Ca2+]i imaging fluorometry. Cells were exposed for 1 minute to APC, protein C zymogen, or mutant Ser360Ala-APC, and [Ca2+]i was monitored in the presence or absence of antibodies against PAR-1, PAR 2, PAR-3, or EPCR. APC, but not protein C zymogen or the active site mutant Ser360Ala-APC, induced dose-dependent [Ca2+]i release in human BECs (Delta[Ca2+]i max = 278.3 +/- 19.5 nM; EC50 for APC = 0.23 +/- 0.02 nM, n = 70 measurements). APC-induced [Ca2+]i signaling was abolished by a cleavage site blocking anti-PAR 1 antibody, whereas anti-PAR-2 and -PAR-3 antibodies were without effect. Antibody RCR252 that ablates APC binding to EPCR blocked APC-mediated [Ca2+]i signaling, whereas anti-EPCR antibody RCR92 that does not block APC binding did not abolish the APC-induced [Ca2+]i response. Experiments using HUVECs confirmed the findings for BECs. Thapsigargin inhibited the APC-induced [Ca2+]i signal, implicating the endoplasmic reticulum as a major source for the APC-induced [Ca2+]i release. These data suggest that APC regulates [Ca2+]i in human brain endothelium and in HUVECs by binding to EPCR and signaling via PAR-1. PMID- 12586612 TI - Chronic lymphocytic leukemias utilizing the VH3-21 gene display highly restricted Vlambda2-14 gene use and homologous CDR3s: implicating recognition of a common antigen epitope. AB - The immunoglobulin variable heavy chain (IgVH) gene mutation status is an important prognostic factor in chronic lymphocytic leukemia (CLL), since cases with mutated VH genes show significantly longer survival than unmutated cases. Recently, we reported a preferential use of the VH3-21 gene in mutated CLL and showed that mutated VH3-21 cases had an inferior overall survival compared with other mutated CLL. In order to further characterize this subset, we performed VH gene analysis in 265 CLL cases and identified 31 VH3-21 cases (11.7%); 21 cases had mutated and 10 cases unmutated VH genes. Regardless of VH gene mutation status, a poor overall survival was found in the VH3-21 cases with a median survival of 83 months. These survival data confirm that VH3-21 cases do not fit into the general prognostic grouping of mutated and unmutated CLL. A large fraction of VH3-21 cases also demonstrated unique features with shorter lengths of the third complementarity determining region (CDR3) and CDR3s with highly homologous amino acid sequences. Furthermore, the VH3-21 cases showed a striking dominance of lambda light chain expression, and analysis of the Iglambda gene rearrangements revealed highly restricted use of the Vlambda2-14/Jlambda3 genes in the majority of cases. Taken together, our new findings strengthen the suggestion that VH3-21-using cases comprise a new CLL entity, irrespective of VH gene mutation status, and implicate that a common antigen epitope, perhaps of pathogenic significance, is recognized by the highly homologous VH3-21/Vlambda2 14 Ig molecules expressed in individual tumors. PMID- 12586613 TI - Signaling through MHC in transgenic mice generates a population of memory phenotype cytolytic cells that lack TCR. AB - We constructed a chimeric molecule, composed of the T-cell receptor (TCR)-zeta chain fused to the extracellular domains of a prototypical allogeneic major histocompatibility complex (MHC) class I molecule, Dd, to assess whether such a construct could affect Dd allospecific responses in vitro and in vivo. To generate cytotoxic T lymphocytes (CTLs) expressing the construct, Dd-zeta was targeted to lymphocyte populations in transgenic mice by placing its expression under control of the CD2 promoter. In response to ligation of Dd, lymphocytes from transgenic mice expressing high levels of Dd-zeta are activated to proliferate and kill cells binding to Dd, despite the near total loss of CD8+ T cells in these mice. Thus, the Dd-zeta cytolytic cell was found not to be a conventional CD8+ CTL, but rather an unusual T lineage cell (CD3-CD5+Thy1.1+) that lacked alphabeta or gammadelta TCRs, as well as CD4 and CD8 coreceptors, but expressed surface markers strikingly similar to memory CTLs, including CD44, Ly 6C, and CD122. These cells originate in the thymus and potently veto responses to Dd in vitro. Lacking TCRs, these veto cells are unlikely to mediate graft-versus host disease (GVHD) and thus may be useful as a cellular therapy for therapeutic deletion of alloreactive T cells in the settings of graft rejection and GVHD. PMID- 12586614 TI - Performance- and safety-enhanced lentiviral vectors containing the human interferon-beta scaffold attachment region and the chicken beta-globin insulator. AB - Retroviral vectors are the most efficient means of stable gene delivery to hematopoietic stem cells (HSCs). However, transgene expression from retroviral vectors is frequently subject to the negative influence of chromosomal sequences flanking the site of integration. Toward the development of autonomous transgene expression cassettes, we inserted the human interferon-beta scaffold attachment region (IFN-SAR) and the chicken beta-globin 5' DNase I hypersensitive site 4 (5'HS4) insulator both separately and together into a series of self-inactivating (SIN) lentiviral vector backbones. Transduced cells of the human CD34+ hematopoietic progenitor line KG1a-pooled populations as well as individual clones harboring single integrants--were analyzed for reporter expression during culture periods of up to 4 months. Vectors carrying both the 5'HS4 insulator and the IFN-SAR consistently outperformed control vectors without inserts as well as vectors carrying either element alone. The performance of a set of vectors containing the murine stem cell virus long terminal repeat as an internal promoter was subsequently assessed during in vitro monocytic differentiation of transduced primary human CD34+ cord blood cells. Similar to what was observed in the KG1a hematopoietic progenitor cell model, optimal reporter expression in primary monocytes was obtained with the vector bearing both regulatory elements. These findings indicate that the 5'HS4/IFN-SAR combination is particularly effective at maintaining open chromatin domains permissive for high-level transgene expression at early and late stages of hematopoietic development, and thus could be of utility in HSC-directed retroviral vector-mediated gene transfer applications. PMID- 12586615 TI - Sphingosine 1-phosphate is a novel inhibitor of T-cell proliferation. AB - Sphingosine 1-phosphate (S1P) is a pleiotropic lysosphingophospholipid stored and secreted by platelets. Using reverse transcription-polymerase chain reaction and flow cytometric analyses, we determined the expression of S1P receptors (S1P1, S1P3, S1P4, and S1P5) in peripheral blood T cells. T cells were induced to proliferate in the presence of phorbol 12-myristate 13-acetate (PMA) plus ionomycin, anti-CD3 plus anti-CD28, and allogeneic immature or mature dendritic cells. This activity was inhibited by the addition of S1P. Enhanced T-cell proliferation was observed when these cells were stimulated with the same stimuli, but were incubated in serum-free media (SFM). Addition of S1P to SFM inhibited the stimulation of T cells induced by T-cell stimuli, suggesting that S1P is an important inhibitory molecule present in the serum. T-cell proliferation was also inhibited by the addition of dihydrosphingosine 1 phosphate (DHS1P), sphingosine, and ceramide; however, the latter 2 sphingolipids required higher concentrations than S1P. Pretreatment of T cells with pertussis toxin (PTX) blocked the inhibitory effect of S1P on activation with PMA plus ionomycin, but not on activation with anti-CD3 plus anti-CD28. This is corroborated with the down-regulation of S1P1 in T cells stimulated with anti-CD3 plus anti-CD28. Similarly, PTX did not affect the inhibitory effect of S1P on T cell proliferation when dendritic cells were used as stimuli. Further, S1P or DHS1P but not ceramide or sphingosine enhanced rather than decreased secretion of interleukin 2 and interferon gamma by T cells stimulated with anti-CD3 plus anti CD28. These results show differential effects of S1P on polyclonal T-cell proliferation and cytokine secretion. PMID- 12586616 TI - High expression of activation-induced cytidine deaminase (AID) and splice variants is a distinctive feature of poor-prognosis chronic lymphocytic leukemia. AB - In chronic lymphocytic leukemia (CLL), analysis of immunoglobulin heavy chain variable regions for somatic hypermutation identifies 2 prognostic subsets, mutated and unmutated. Investigators have postulated that unmutated and mutated CLL arises from malignant transformation of pre- and post-germinal center (GC) B cells, respectively. Alternatively, unmutated cases may arise from B cells stimulated by T-cell-independent antigens or from GC B cells with inactive somatic hypermutation. Activation-induced cytidine deaminase (AID), a protein essential for somatic hypermutation, is expressed by GC B cells in which this process occurs. We investigated AID mRNA expression in 20 CLL cases. In 8 cases we detected high expression of wild-type AID mRNA and 2 splice variants; in 12 cases and 5 normal peripheral blood B-cell samples we detected no expression using standard conditions. Of 8 CLL cases that highly expressed AID, 7 were unmutated, suggesting that this subset may arise from GC-experienced B cells with inactive somatic hypermutation, and may predict prognosis. PMID- 12586617 TI - Prevalence, causes, and characterization of factor XI inhibitors in patients with inherited factor XI deficiency. AB - Factor XI deficiency, an injury-related bleeding disorder, is rare worldwide but common in Jews in whom 2 mutations, Glu117Stop (type II) and Phe283Leu (type III), prevail. Mean factor XI activities in homozygotes for Glu117Stop and for Phe283Leu are 1 and 10 U/dL, respectively. Inhibitors to factor XI in patients with severe factor XI deficiency have been reported in a small number of instances. This study was undertaken to determine the prevalence of acquired inhibitors against factor XI in patients with severe factor XI deficiency, discern whether these inhibitors are related to specific mutations, and characterize their activity. Clinical information was obtained from unrelated patients with severe factor XI deficiency, and blood was analyzed for factor XI activity, inhibitor to factor XI, and causative mutations. Immunoglobulin G purified from patients with an inhibitory activity was tested for binding to factor XI, effects on activation of factor XI by factor XIIa and thrombin, and activation of factor IX by exogenous factor XIa. Of 118 Israeli patients, 7 had an inhibitor; all belonged to a subgroup of 21 homozygotes for Glu117Stop who had a history of plasma replacement therapy. Three additional patients with inhibitors from the United Kingdom and the United States also had this genotype and were exposed to plasma. The inhibitors affected factor XI activation by thrombin or factor XIIa, and activation of factor IX by factor XIa. The results imply that patients with a very low factor XI level are susceptible to development of an inhibitor following plasma replacement. PMID- 12586618 TI - Minimal residual disease in adolescent (older than 14 years) and adult acute lymphoblastic leukemias: early immunophenotypic evaluation has high clinical value. AB - Investigation of minimal residual disease (MRD) in acute leukemias by immunophenotyping and/or molecular techniques is proving to be increasingly valuable for disease monitoring. In acute lymphoblastic leukemia (ALL), most MRD studies have focused on children, whereas in contrast, information on the value of MRD on adult ALL is scanty, and almost exclusively restricted to polymerase chain reaction (PCR) studies. Early response to therapy is one of the most important prognostic factors in acute leukemia, which prompted us to investigate whether or not early immunophenotypic assessment of MRD could also be a valuable tool for predicting relapse in adult patients with ALL. For that purpose we have analyzed the level of MRD during the initial phase of treatment (induction phase) by multiparameter flow cytometry in a series of 102 adolescent (older than 14 years) and adult patients with ALL. Immunophenotypic evaluation of the bone marrow (BM) at day +35 showed that patients with low MRD levels (< 0.05% leukemia associated phenotype [LAP+] cells) had a significantly longer relapse-free survival (RFS) than patients with high MRD levels, and this prognostic influence was retained when only those patients in morphologic complete remission (mCR) at day +35 were considered (median RFS: 42 months vs 16 months; P =.001). Moreover, immunophenotyping helped to identify a small subset of patients (n = 12) with negative or low MRD levels (< 0.03% LAP+ cells) by day +14, with an excellent prognosis (projected RFS of 90% at 5 years). The contrary is true of patients who achieved late mCR (after day +35), since immunophenotypic investigation of MRD showed that, in spite of the mCR, none of the cases with more than 0.1% LAP+ cells would be relapse-free after 2 years. Multivariate analysis showed that the immunologic evaluation of MRD at day +35 was the most relevant independent prognostic parameter for adult patients with ALL, and together with age, white blood cell (WBC) count at diagnosis, and presence of the Philadelphia (Ph) chromosome, represented the most informative combination of variables for predicting relapse-free survival. PMID- 12586619 TI - Aberrant DNA methylation of p57KIP2 identifies a cell-cycle regulatory pathway with prognostic impact in adult acute lymphocytic leukemia. AB - P57KIP2 is a cyclin-dependent kinase inhibitor silenced in a variety of human malignancies. DNA methylation of a region surrounding the transcription start site of p57KIP2 was found in acute lymphocytic leukemia (ALL)-derived cell lines. Methylation of this region correlated with gene silencing, and treatment of methylated/silenced cell lines with 5-aza-2'-deoxycytidine resulted in gene re expression. P57KIP2 was methylated in 31 (50%) of 63 patients with newly diagnosed ALL, and in 11 (52%) of 21 patients with relapsed ALL. In 5 of them (25%), methylation was acquired at relapse. No association was observed between methylation of p57KIP2 alone and clinical-biologic characteristics studied, including overall survival (OS) or disease-free survival. Methylation of multiple genes in a cell-cycle regulatory pathway composed of p73, p15, and p57KIP2 occurred in 22% of Philadelphia chromosome (Ph)-negative patients. Ph-negative patients with methylation of 2 or 3 genes of this pathway had a significantly worse median OS compared with those with methylation of 0 or 1 gene (50 vs 467 weeks, respectively; P =.02). Our results indicate that p57KIP2 is frequently methylated in adult patients with ALL, and that inactivation of a pathway composed of p73, p15, and p57KIP2 predicts for poor prognosis in Ph-negative patients. PMID- 12586620 TI - GATA1 mutations in transient leukemia and acute megakaryoblastic leukemia of Down syndrome. AB - Children with constitutional trisomy 21 (Down syndrome) have an approximately 500 fold increased risk of developing acute megakaryoblastic leukemia (AMKL), a form of acute myeloid leukemia. Unique to newborn infants with Down syndrome is a transient leukemia (TL), also referred to as transient myeloproliferative syndrome, that undergoes spontaneous remission in the majority of cases but in approximately 20% is followed by AMKL later in life. Recently mutations of the gene encoding the hematopoietic transcription factor GATA1 were shown to be specific for AMKL of Down syndrome. Here, we demonstrate that GATA1 mutations are present in blasts of TL and show the identical GATA1 mutation in sequential samples collected from a patient during TL and subsequent AMKL. These findings suggest a model of malignant transformation in Down syndrome AMKL in which GATA1 mutations are an early event and AMKL arises from latent TL clones following initial apparent remission. PMID- 12586621 TI - Unrelated cord blood transplantation for adult patients with advanced myelodysplastic syndrome. AB - We report the results of unrelated cord blood transplantation (CBT) for 13 adult patients with advanced myelodysplastic syndrome (MDS). The median age was 40 years, the median weight was 51 kg, and the median number of infused nucleated cells was 2.43 x 107/kg. Twelve patients had myeloid reconstitution, and the median time to more than 0.5 x 109/L (5 x 108/L) absolute neutrophil count was 22.5 days. A self-sustained platelet count more than 50 x 109/L was achieved in 11 patients at a median time of 49 days. Acute graft versus host disease (GVHD) occurred in 9 of 12 evaluable patients and chronic GVHD in 8 of 11 evaluable patients. Ten patients are alive and free of disease at between 171 and 1558 days after transplantation. The probability of disease-free survival at 2 years was 76.2%. These results suggest that adult advanced MDS patients without suitable related or unrelated bone marrow donors should be considered as candidates for CBT. PMID- 12586622 TI - Human cytomegalovirus immediate-early mRNAemia versus pp65 antigenemia for guiding pre-emptive therapy in children and young adults undergoing hematopoietic stem cell transplantation: a prospective, randomized, open-label trial. AB - In the search for better protocols of preemptive therapy of human cytomegalovirus (HCMV) infection in hematopoietic stem cell transplant (HSCT) recipients, we conducted a randomized trial comparing antigenemia with the nucleic acid sequence based assay (NASBA) for determination of HCMV immediate-early messenger RNA (IEmRNA) as the guiding assay for initiation of pre-emptive antiviral treatment. In the IEmRNA arm, antiviral therapy was started upon IEmRNA positivity confirmed the following day, whereas in the antigenemia arm, therapy was started in the presence of either at least 2 pp65-positive leukocytes/2 x 105 examined or a single positive leukocyte confirmed the following day. In both arms, treatment was stopped upon 2 consecutive negative results. All patients were monitored for 3 months after HSCT. The primary end point of the study was duration of anti-HCMV therapy. On the whole, 80 children (41 in the IEmRNA and 39 in the antigenemia arm), recipients of transplants from either a relative or an unrelated donor, completed the study. No patient developed HCMV disease. In the IEmRNA arm, the incidence of HCMV infection was higher compared to the antigenemia arm (80% vs 51%, respectively, P =.0069), as well as the percentage of treated patients (66% vs 44%, respectively, P =.045). However, the percentage of relapses and treated relapses was comparable in the 2 arms. There was no significant difference in median duration of therapy per patient. Although these data indicate that IEmRNA determination does not offer advantages in terms of treatment duration, it can safely replace antigenemia, while semiautomation is the major advantage of the NASBA procedure. PMID- 12586623 TI - Mechanisms and implications of platelet discoid shape. AB - The platelet marginal band consists of a single peripheral microtubule (MT) that is wound in 8 to 12 coils and maintains discoid cell shape. About 90% of beta tubulin in the marginal band is of the divergent, megakaryocyte (MK)/platelet restricted beta1 isoform. beta1-tubulin-null mice show reduced proplatelet formation, thrombocytopenia, and platelet spherocytosis. Here, we show that structural abnormalities in resting beta1-tubulin-/- platelets include frequent kinks and breaks in the marginal band. Platelets derived from mice lacking the transcription factor GATA1 show similar defects, probably as a direct consequence of absent beta1-tubulin. beta1-tubulin+/- platelets have normal ratios of beta tubulin isotypes but the marginal band is half the normal thickness, which is sufficient to maintain elliptical cell shape. Thus, a threshold 50% or less of the normal amount of beta1-tubulin is required to preserve marginal band integrity and cell shape. beta1-tubulin-/- platelets have normal size and contents and show no defects in serotonin release or aggregation. Accordingly, the apparently isolated spherocytosis allows investigation of the role of discoid platelet shape in hemostasis. On agonist stimulation, the disorganized MTs in beta1-tubulin-/- platelets fail to condense into central rings and instead are dispersed in short bundles and linear arrays. Nevertheless, intravital microscopy and flow chamber studies demonstrate full functionality of these spherocytic platelets under physiologic shear conditions. Together, these findings highlight the essential requirements of the MK/platelet-restricted beta1-tubulin isoform in platelet structure and suggest that spherocytosis does not impair many aspects of platelet function. PMID- 12586624 TI - IL-15 is an essential mediator of peripheral NK-cell homeostasis. AB - Several distinct classes of surface receptors can, on ligand binding, transmit signals that modulate the survival, proliferation, and apoptosis of peripheral B, T, and natural killer (NK) cells. At the population level, dynamic changes in lymphocyte cell numbers are strictly regulated to maintain a steady state, a process referred to as homeostasis. Although several studies have investigated the signals that regulate B- and T-cell homeostasis, little is known about the mechanisms that control the survival and proliferation of peripheral NK cells. Using an adoptive transfer system, we have investigated the role of gammac dependent cytokines, in particular interleukin 7 (IL-7) and IL-15, and major histocompatibility complex (MHC) class I molecules in peripheral NK-cell homeostasis. We observed that IL-15 plays a dominant role in the survival of peripheral NK cells, via maintenance of the antiapoptotic factor Bcl-2. IL-15 availability, however, also plays an important role because endogenous NK cells in the recipient mice influence the behavior of adoptively transferred NK cells. Finally, although NK cells bear functional inhibitory Ly49 receptors for MHC class I molecules, the presence or absence of specific ligands on host cells did not influence the survival or homeostatic expansion of donor NK cells. PMID- 12586625 TI - Specific homeodomain-DNA interactions are required for HOX11-mediated transformation. AB - HOX11 encodes a homeodomain protein that is aberrantly expressed in T-cell acute lymphoblastic leukemia as a consequence of the t(10;14) and t(7;10) chromosomal translocations. We previously reported that HOX11 immortalizes murine hematopoietic progenitors and induces pre-T-cell tumors in mice after long latency. It has been demonstrated in a number of studies that HOX11, similar to other homeodomain proteins, binds DNA and transactivates transcription. These findings suggest that translocation-activated HOX11 functions as an oncogenic transcription factor. Here we report that HOX11 represses transcription through both TATA-containing and TATA-less promoters. Interestingly, transcriptional repression by HOX11 is independent of its DNA binding capability. Moreover, a systematic mutational analysis indicated that repressor activity was separable from immortalizing function, which requires certain residues within the HOX11 homeodomain that make base-specific or phosphate-backbone contacts with DNA. We further showed that the pathologic action of HOX11 involves DNA binding-dependent transcriptional pathways that are distinct from those controlling expression of a chromosomal target gene (Aldh-1). We conclude that dysregulated expression of a particular set of downstream target genes by DNA binding via the homeodomain is of central importance for leukemia initiation mediated by HOX11. PMID- 12586626 TI - Synergy between all-trans retinoic acid and tumor necrosis factor pathways in acute leukemia cells. AB - The nuclear receptor ligand all-trans retinoic acid (ATRA) causes dramatic terminal differentiation of acute promyelocytic leukemia (APL) cells in vitro and in patients, but it is less active in other malignancies. However, downstream mediators of the effects of ATRA are not well understood. We used a cDNA microarray to search for ATRA-regulated genes in the APL cell line NB4 and found that ATRA regulated several members of the tumor necrosis factor (TNF) pathway. Here we show that TNF can synergize with ATRA to induce differentiation, showing monocytic characteristics more typical of differentiation mediated by TNF than by ATRA. ATRA and TNF can also induce differentiation of the non-APL cell line U937. Underlying this response was an increase in TNF-induced nuclear factor-kappaB (NF kappaB) DNA binding within 2 hours in the presence of ATRA and activation of NF kappaB DNA binding and transcriptional activity in response to ATRA alone within 48 hours of ATRA treatment. Furthermore, we found a synergistic induction of the NF-kappaB target genes BCL-3, Dif-2, and TNF receptor 2 (TNFR2) in response to the combination of TNF and ATRA. These genes have been previously shown to play a role in TNF signaling, and amplification of such genes may represent a mechanism whereby TNF and ATRA can act synergistically. We propose that ATRA can prime cancer cells for differentiation triggered by TNF and suggest that targeting the TNF pathway in combination with ATRA may represent a novel route to treat leukemias. PMID- 12586627 TI - Oxidized low-density lipoprotein (oxLDL) triggers hypoxia-inducible factor-1alpha (HIF-1alpha) accumulation via redox-dependent mechanisms. AB - Oxidized low-density lipoprotein (oxLDL) and macrophages play a central role in atherosclerosis. Here, we obtained evidence that oxLDL induced hypoxia-inducible factor-1alpha (HIF-1alpha) protein accumulation in human macrophages (Mono-Mac-6) under normoxia. HIF-1alpha accumulation was attenuated by pretreatment with the antioxidant N-acetyl-L-cysteine (NAC), the nitric oxide (NO) donor S nitrosoglutathione (GSNO), and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitors such as diphenyleniodonium (DPI) or 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF), thus implicating the contribution of oxLDL generated reactive oxygen species (ROS). Whereas oxLDL did not modulate HIF 1alpha mRNA levels, experiments with cycloheximide pointed to a translational mechanism in oxLDL action. HIF-1-dependent luciferase reporter gene analysis underscored HIF-1 transactivation. Our results indicate that oxLDL induced HIF 1alpha accumulation and HIF-1-dependent reporter gene activation in human macrophages via a redox-mediated pathway. This finding may suggest a role of HIF 1 in atherosclerosis and oxLDL-induced pathogenesis. PMID- 12586628 TI - Rituximab in lymphocyte-predominant Hodgkin disease: results of a phase 2 trial. AB - Lymphocyte-predominant Hodgkin disease (LPHD) is a unique clinical entity characterized by indolent nodal disease that tends to relapse after standard radiotherapy or chemotherapy. The malignant cells of LPHD are CD20+ and therefore rituximab may have activity with fewer late effects than standard therapy. In this phase 2 trial, 22 patients with CD20+ LPHD received 4 weekly doses of rituximab at 375 mg/m2. Ten patients had previously been treated for Hodgkin disease, while 12 patients had untreated disease. All 22 patients responded to rituximab (overall response rate, 100%) with complete response (CR) in 9 (41%), unconfirmed complete response in 1 (5%), and partial response in 12 (54%). Acute treatment-related adverse events were minimal. With a median follow-up of 13 months, 9 patients had relapsed, and estimated median freedom from progression was 10.2 months. Progressive disease was biopsied in 5 patients: 3 had recurrent LPHD, while 2 patients had transformation to large-cell non-Hodgkin lymphoma (LCL). All 3 patients with recurrent LPHD were retreated with rituximab, with a second CR seen in 1 patient and stable disease in 2. Rituximab induced prompt tumor reduction in each of 22 LPHD patients with minimal acute toxicity; however, based on the relatively short response duration seen in our trial and the concerns about transformation, rituximab should be considered investigational treatment for LPHD. Further clinical trials are warranted to determine the optimal dosing schedule of rituximab, the potential for combination treatment, and the possible relationship of rituximab treatment to the development of LCL. PMID- 12586629 TI - Peroxiredoxin II is essential for sustaining life span of erythrocytes in mice. AB - Peroxiredoxins (Prxs) are a family of antioxidant proteins that reduce peroxide levels by using reducing agents such as thioredoxin. These proteins were characterized to have a number of cellular functions, including cell proliferation and differentiation and protection of specific proteins from oxidative damage. However, the physiological roles of the peroxiredoxins have not been determined. To clarify the physiological relevance of this protein type, we generated a mouse model deficient in Prx II, which is abundantly expressed in all types of cells. The Prx II-/- mice were healthy in appearance and fertile. However, they had splenomegaly caused by the congestion of red pulp with hemosiderin accumulation. Heinz bodies were detected in their peripheral blood, and morphologically abnormal cells were elevated in the dense red blood cell (RBC) fractions, which contained markedly higher levels of reactive oxygen species (ROS). The Prx II-/- mice had significantly decreased hematocrit levels, but increased reticulocyte counts and erythropoietin levels, indicative of a compensatory action to maintain hematologic homeostasis in the mice. In addition, a labeling experiment with the thiol-modifying reagent biotinylated iodoacetamide (BIAM) in Prx II-/- mice revealed that a variety of RBC proteins were highly oxidized. Our results suggest that Prx II-/- mice have hemolytic anemia and that Prx II plays a major role in protecting RBCs from oxidative stress in mice. PMID- 12586630 TI - Platelet factor 4 promotes adhesion of hematopoietic progenitor cells and binds IL-8: novel mechanisms for modulation of hematopoiesis. AB - Platelet factor 4 (PF4) is an abundant platelet alpha-granule C-X-C chemokine that has weak chemotactic potency but strongly inhibits hematopoiesis through an unknown mechanism. We find that PF4 binds to human CD34+ hematopoietic progenitor cells (HPCs) with a median effective concentration of 1 microg/mL but not after exposure to chondroitinase ABC. PF4 enhances adhesion of HPCs to intact stroma. Committed progenitors also adhere avidly to immobilized PF4. This adhesion is time-dependent, requires metabolic activity, causes cytoskeletal rearrangement, and induces cell-cycle inhibition. Using extracellular acidification rate to indicate transmembrane signaling, we find that interleukin-8 (IL-8), but not PF4, activates CD34+ progenitors, and PF4 blocks IL-8-mediated activation. Surface plasmon resonance analysis shows that PF4 binds IL-8 with high (dissociation constant [Kd] = 42 nM) affinity. Nuclear magnetic resonance analysis of IL-8 and PF4 in solution confirms this interaction. We conclude that PF4 has the capacity to influence hematopoiesis through mechanisms not mediated by a classical high affinity, 7-transmembrane domain chemokine receptor. Instead, PF4 may modulate the hematopoietic milieu both directly, by promoting progenitor adhesion and quiescence through interaction with an HPC chondroitin sulfate-containing moiety, and indirectly, by binding to or interfering with signaling caused by other, hematopoietically active chemokines, such as IL-8. PMID- 12586631 TI - The carboxy-terminal region of the granulocyte colony-stimulating factor receptor transduces a phagocytic signal. AB - Granulocyte colony-stimulating factor (G-CSF) induces proliferation, maturation, and functional activities of myeloid progenitors and mature neutrophils through a specific receptor, the G-CSF-R. Different signals are mediated by distinct regions of the cytoplasmic domain of G-CSF-R, but the precise role of each region has not yet been fully clarified. We evaluated the involvement of Syk kinase, essential in mediating phagocytic signals by Fcgamma receptors, in G-CSF-induced phagocytosis, using murine myeloid 32D cells transfected with wild-type (WT) human G-CSF-R (hG-CSF-R) or with a G-CSF-R mutant truncated at cytoplasmic amino acid 715. The G-CSF-R mutant lacks the immunoreceptor tyrosine-based activation motif (ITAM), putative binding site for Syk. Following treatment of WT hG-CSF-R transfectants with IgG-coated particles, there was a significant increase in phagocytosis in G-CSF-stimulated cells, in which Syk tyrosine phosphorylation occurred, paralleled by enhancement of its tyrosine kinase activity. In the mutant transfectants, no significant increase in phagocytosis or Syk tyrosine phosphorylation occurred after stimulation with G-CSF. We also demonstrated that tyrosine phosphorylation of the Src kinases Hck and Lyn occurs following G-CSF stimulation of cells expressing WT G-CSF-R, but that Hck is not phosphorylated in mutant G-CSF-R transfectants. The increase in phagocytosis following G-CSF stimulation cannot be attributed to a rapid de novo increase in expression of Fcgamma receptors. G-CSF induced expression of Fcgamma receptors only after prolonged stimulation. Our data provide evidence that the carboxy-terminal region of G-CSF-R plays a role in the phagocytosis of IgG-coated particles and that Syk and Hck kinase tyrosine phosphorylation is involved. PMID- 12586632 TI - Ectopic hTERT expression extends the life span of human CD4+ helper and regulatory T-cell clones and confers resistance to oxidative stress-induced apoptosis. AB - Human somatic cells have a limited life span in vitro. Upon aging and with each cell division, shortening of telomeres occurs, which eventually will lead to cell cycle arrest. Ectopic hTERT expression has been shown to extend the life span of human T cells by preventing this telomere erosion. In the present study, we have shown that ectopic hTERT expression extends the life span of CD4+ T helper type 1 or 2 and regulatory T-cell clones and affected neither the in vitro cytokine production profile nor their specificity for antigen. In mixed cell cultures, ectopic hTERT-expressing clones were found to expand in greater numbers than untransduced cells of the same replicative age. This ectopic hTERT-induced growth advantage was not due to an enhanced cell division rate or number of divisions following T-cell receptor-mediated activation, as determined in carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeling experiments. Moreover, the susceptibility to activation-induced cell death of both cell types was similar. However, cultures of resting hTERT-transduced T cells contained higher frequencies of Bcl-2-expressing cells and lower active caspase-3 expressing cells, compared with wild-type cells. Furthermore, hTERT-transduced cells were more resistant to oxidative stress, which causes preferential DNA damage in telomeres. Taken together, these results show that ectopic hTERT expression not only protects proliferating T cells from replicative senescence but also confers resistance to apoptosis induced by oxidative stress. PMID- 12586633 TI - VEGF inhibits T-cell development and may contribute to tumor-induced immune suppression. AB - T-cell defects and premature thymic atrophy occur in cancer patients and tumor bearing animals. We demonstrate that exposure of mice to recombinant vascular endothelial growth factor (VEGF) at concentrations similar to those observed in advanced stage cancer patients reproduces this profound thymic atrophy and is highlighted by a dramatic reduction in CD4+/CD8+ thymocytes. We find that VEGF does not induce thymocyte apoptosis, but instead rapidly decreases the number of the earliest observable progenitors in the thymus. VEGF does not inhibit thymocyte development in fetal thymic organ culture, further suggesting a prethymic effect. We also demonstrate that bone marrow progenitors from animals infused with recombinant VEGF and transferred to irradiated untreated animals recolonize the thymus more efficiently than progenitors from control animals. This suggests that VEGF exposure is associated with an increased population of thymus-committed progenitors in the bone marrow. We hypothesize that pathophysiologically relevant concentrations of VEGF may block the differentiation and/or emigration of these progenitors resulting in the observed thymic atrophy. Removal of VEGF via cessation of infusion or adoptive transfer of progenitors to a congenic host induces a preferential commitment of lymphoid progenitors to the T lineage and results in a restoration of the normal composition and cellularity of the thymus. These data demonstrate that at pathophysiologic concentrations, VEGF interferes with the development of T cells from early hematopoetic progenitor cells and this may contribute to tumor associated immune deficiencies. PMID- 12586634 TI - Host stem cells can selectively reconstitute missing lymphoid lineages in irradiation bone marrow chimeras. AB - The regulatory elements governing the process of lymphopoiesis from pluripotential stem cells to mature lymphocytes are not well understood. In this study we found that in bone marrow chimeras made by reconstituting lethally irradiated normal mice with bone marrow taken from genetically B-cell-deficient animals (microMT.B6 --> F1) the B-cell compartment is reconstituted with host derived B cells. Similarly, in animals reconstituted with bone marrow taken from mice with genetic deficiencies in the development of T cells (TCR-/- --> F1) or both B and T cells (RAG-/- --> F1), the missing lymphocyte lineage(s) was specifically reconstituted from host-derived cells. In all chimeras, all other blood lineages were generated from donor-derived stem cells. Control chimeras (B6 --> F1) had only donor-derived hematopoietic cells as expected. The reconstituted, host-derived lymphoid compartments contained normal functional cell populations as determined by the presence of T cells expressing all 16 common TCR Vbeta families, and the presence of all antibody isotypes in the serum. Reconstituted TCR-/- --> F1 chimeras were also able to mount T-cell proliferative responses to foreign antigens equal to those of control animals. This observation would seem to suggest that during lymphopoietic reconstitution, missing lymphoid lineages can dictate their own reconstitution. PMID- 12586635 TI - The resistance of B-CLL cells to DNA damage-induced apoptosis defined by DNA microarrays. AB - B-cell chronic lymphoid leukemia (BCLL) is a highly heterogeneous human malignancy, presumably reflecting specific molecular alterations in gene expression and protein activity that are thought to underlie the variable disease outcome. Most B-CLL cell samples undergo apoptotic death in response to DNA damage. However, a clinically distinct aggressive subset of B-CLL is completely resistant in vitro to irradiation-induced apoptosis. We therefore addressed 2 series of microarray analyses on 4 sensitive and 3 resistant B-CLL cell samples and compared their gene expression patterns before and after apoptotic stimuli. Data analysis pointed out 16 genes whose expression varied at least 2-fold specifically in resistant cells. We validated these selected genes by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) on 7 microarray samples and confirmed their altered expression level on 15 additional B-CLL cell samples not included in the microarray analysis. In this manner, in 11 sensitive and 11 resistant B-CLL cell samples tested, 13 genes were found to be specific for all resistant samples: nuclear orphan receptor TR3, major histocompatibility complex (MHC) class II glycoprotein HLA-DQA1, mtmr6, c-myc, c rel, c-IAP1, mat2A, and fmod were up-regulated, whereas MIP1a/GOS19-1 homolog, stat1, blk, hsp27, and ech1 were down-regulated. In some cases, the expression profile may be dependent on the status of p53. Some of these genes encode general apoptotic factors but also exhibit lymphoid cell specificities that could potentially be linked to the development of lymphoid malignancies (MIP1alpha, blk, TR3, mtmr6). Taken together, our data define new molecular markers specific to resistant B-CLL subsets that might be of clinical relevance. PMID- 12586637 TI - Phasic contractions of the rat portal vein depend on intracellular Ca2+ release stimulated by depolarization. AB - The phasic contraction to phenylephrine of the rat isolated portal vein was investigated using functional studies. Phasic contractions to phenylephrine and caffeine could be produced after several minutes in Ca(2+)-free Krebs solution, which were inhibited by cyclopiazonic acid or ryanodine. The phenylephrine and caffeine contractions were abolished, however, within 10 min in Ca(2+)-free Krebs solution and by nifedipine. This indicated the Ca(2+) stores were depleted in the absence of Ca(2+) influx through voltage-gated channels. The phasic contraction to phenylephrine was also abolished by niflumic acid even in Ca(2+)-free Krebs solution. This showed that the response depended on intracellular Ca(2+) release stimulated directly by depolarization, resulting from opening of Ca(2+)-activated Cl(-) channels, but did not require Ca(2+) influx. In support of this, K(+) induced phasic contractions were also produced in Ca(2+)-free Krebs solution. The phenylephrine but not K(+)-induced phasic contractions in Ca(2+)-free Krebs solution were inhibited by ryanodine or cyclopiazonic acid. This would be consistent with Ca(2+) release from more superficial intracellular stores (affected most by these agents), probably by inositol 1,4,5-trisphospate, being required to stimulate the phenylephrine depolarization. PMID- 12586636 TI - ACE and non-ACE mediated effect of angiotensin I on intracellular calcium mobilization in rat glomerular arterioles. AB - Because renin and angiotensin I (ANG I) level are high in the renal circulation, the conversion of ANG I is a critical step in the regulation of glomerular hemodynamics. We studied this conversion by investigating the effect of ANG I on intracellular Ca(2+) concentration ([Ca(2+)](i)) in rat juxtamedullary glomerular afferent and efferent arterioles (AA and EA, respectively). Two types of EA were considered, thin EA and muscular EA, terminating as peritubular capillaries and vasa rectae, respectively. In all arterioles, ANG I elicited [Ca(2+)](i) elevations. Maximal responses of 171 +/- 28 (AA), 183 +/- 7 (muscular EA), and 78 +/- 11 nM (thin EA) (n = 6), similar to those obtained with ANG II, were observed with 100 nM ANG I. The EC(50) values were 20 times higher for ANG I than for ANG II in AA (10.2 vs. 0.5) and muscular EA (6.8 vs. 0.4 nM) and 150 times higher in thin EA (15.2 vs. 0.1 nM). ANG I effect was blocked by losartan, indicating that AT(1) receptors were involved. The ANG-converting enyzme (ACE) inhibitor lisinopril inhibited the maximal response to ANG I in AA and muscular EA by 75 +/ 9% (n = 13) and 70 +/- 7% (n = 13), respectively, but had no effect in thin EA (n = 14). The serine protease inhibitor aprotinin, the chymase inhibitor chymostatin, and the cysteine protease inhibitors E64 and leupeptin had no effect on ANG I action. These data show that ANG I effects are mainly mediated by ACE in AA and muscular EA but not in thin EA. The lisinopril-insensitive response may be related to conversion by unknown enzyme(s) and/or to activation of AT(1) receptors by ANG I. PMID- 12586638 TI - Effect of ischemia on soluble and particulate guanylyl cyclase-mediated cGMP synthesis in cardiomyocytes. AB - The effect of simulated ischemia [hypoxia, no glucose, extracellular pH (pH(o)) 6.4] on cGMP synthesis induced by stimulation of soluble (sGC) or particulate guanylyl cyclase (pGC) was investigated in adult rat cardiomyocytes. Intracellular cGMP content was measured after stimulation of sGC by S-nitroso-N penicillamine (SNAP) or stimulation of pGC by natriuretic peptides [urodilatin (Uro), atrial natriuretic peptide (ANP), or C-type natriuretic peptide (CNP)] for 1 min in the presence of phosphodiesterase inhibitors. After 2 h of simulated ischemia, a decrease of >50% was observed in pGC-dependent cGMP synthesis, but no significant change was observed in sGC-dependent cGMP synthesis. The reduction in cGMP synthesis caused by simulated ischemia was mimicked by extracellular acidosis (pH(o) 6.4), which decreased pGC-mediated cGMP synthesis without altering sGC-mediated cGMP synthesis. An extreme sensitivity of pGC activity to low pH was also observed in membrane cell fractions. Hypoxia without acidosis (pH(o) 7.4) profoundly depressed cellular ATP content but did not change the response to SNAP, Uro, or ANP (selective agonists of pGC type A receptor). Only cGMP synthesis in response to CNP (a selective agonist of pGC type B receptor) was significantly reduced by ATP depletion. These data support the relevance of intracellular pH as a modulator of cGMP and suggest that, in ischemic cardiomyocytes, synthesis of cGMP would be mainly nitric oxide dependent. PMID- 12586639 TI - Metalloendopeptidases EC 3.4.24.15/16 regulate bradykinin activity in the cerebral microvasculature. AB - Bradykinin is a vasoactive peptide that has been shown to increase the permeability of the cerebral microvasculature to blood-borne macromolecules. The two zinc metalloendopeptidases EC (EP 24.15) and EC (EP 24.16) degrade bradykinin in vitro and are highly expressed in the brain. However, the role that these enzymes play in bradykinin metabolism in vivo remains unclear. In the present study, we investigated the role of EP 24.15 and EP 24.16 in the regulation of bradykinin-induced alterations in microvascular permeability. Permeability of the cerebral microvasculature was assessed in anesthetized Sprague-Dawley rats by measuring the clearance of 70-kDa FITC dextran from the brain. Inhibition of EP 24.15 and EP 24.16 by the specific inhibitor N-[1-(R,S)-carboxy-3-phenylpropyl] Ala-Aib-Tyr-p-aminobenzoate (JA-2) resulted in the potentiation of bradykinin induced increases in cerebral microvessel permeability. The level of potentiation was comparable to that achieved by the inhibition of angiotensin-converting enzyme. These findings provide the first evidence of an in vivo role for EP 24.15/EP 24.16 in brain function, specifically in regulating alterations in microvessel permeability induced by exogenous bradykinin. PMID- 12586640 TI - Chronic effects of angiotensin-converting enzyme inhibition on kinin receptor binding sites in the rat spinal cord. AB - With the use of in vitro receptor autoradiography, this study aims at determining whether the higher level of kinin B(2) receptor density in the spinal cord of the spontaneously hypertensive rat (SHR) is secondary to arterial hypertension and whether chronic treatment with angiotensin I-converting enzyme inhibitors (ACEI) can regulate neuronal B(1) and B(2) receptors. SHR received, from the age of 4 wk, one of the two ACEI (lisinopril or zofenopril, 10 mg x kg(-1) x day(-1)) or for comparison, the selective AT(1) antagonist (losartan, 20 mg x kg(-1) x day( 1)) in their drinking water for a period of 4, 12, and 20 wk. Age-matched untreated SHR and Wistar-Kyoto rats (WKY) were used as controls. B(2) receptor binding sites in most laminae were higher in SHR than in WKY from the age of 8 to 24 wk. Whereas B(1) receptor binding sites were significantly present in young SHR and WKY, they were barely detectable in adult rats. ACEI (16 and 24 wk) and AT(1) antagonist (24 wk) enhanced the number of B(2) without changing B(1) receptor binding sites. However, at 8 wk the three treatments significantly increased B(1) and decreased B(2) receptors in lamina I. It is concluded that 1) the higher density of B(2) receptors in the spinal cord of SHR is not due to hypertension, 2) kinin receptors are regulated differently by ACEI in neuronal and vascular tissues, and 3) aging may have a profound impact on levels of B(1) and B(2) receptors in the rat spinal cord. PMID- 12586641 TI - Intravital microscopy comparing T lymphocyte trafficking to the spleen and the mesenteric lymph node. AB - Lymphocyte rolling velocity is determined largely by interactions between leukocyte alpha(4)-integrin (CD49d) and L-selectin and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in mesenteric postcapillary venules and Peyer's patch high endothelial venules (HEVs). The role of these interactions in other tissue sites of lymphocyte emigration is not known. With the use of real-time intravital confocal microscopy, we found that rolling velocities of T lymphocytes in the murine mesenteric lymph node (MLN) HEV also depend on L-selectin and CD49d. However, in the murine spleen, rolling velocities of T lymphocytes are not influenced by the loss of L-selectin and CD49d. With the use of FITC-dextran and TIE2-GFP mice, we further defined the microvascular compartments of the spleen and showed that adherence of T cells is localized to regions in the white pulp that are not lined by endothelial cells and have shear rates similar to bone marrow sinusoids. These results establish that T cell trafficking to the spleen differs from trafficking to other secondary lymphoid organs and suggest that the mechanical properties of the blood-filtering role of the spleen are important in T cell accumulation in the organ. PMID- 12586642 TI - Congestion charging and the walking classes. PMID- 12586643 TI - New guidelines on the management of asthma. PMID- 12586644 TI - Safety and efficacy of interventional procedures. PMID- 12586645 TI - Not to be taken as directed. PMID- 12586646 TI - Sudden death in the shadows of epilepsy. PMID- 12586647 TI - US seniors group attacks pharmaceutical industry "fronts". PMID- 12586648 TI - United Kingdom finally bans tobacco advertising. PMID- 12586651 TI - US given more time to consider cheap drugs deal for poor nations. PMID- 12586653 TI - Initiative will fast-track vaccine for childhood diarrhoea in developing world. PMID- 12586654 TI - India launches massive polio vaccine programme. PMID- 12586659 TI - Women fail to recognise risk of heart disease. PMID- 12586665 TI - Threat of war is affecting mental health of Iraqi children, says report. PMID- 12586666 TI - Short and long term mortality associated with foodborne bacterial gastrointestinal infections: registry based study. AB - OBJECTIVES: To determine the excess mortality associated with infections with Salmonella, Campylobacter, Yersinia enterocolitica, and Shigella and to examine the effect of pre-existing illness. DESIGN: Registry based, matched cohort study. SETTING: Denmark. PARTICIPANTS: 48 857 people with gastrointestinal infections plus 487 138 controls from the general population. MAIN OUTCOME MEASURE: One year mortality among patients with gastrointestinal infections compared with controls after adjustment for comorbidity. RESULTS: 1071 (2.2%) people with gastrointestinal infections died within one year after infection compared with 3636 (0.7%) controls. The relative mortality within one year was 3.1 times higher in patients than in controls. The relative mortality within 30 days of infection was high in all four bacterial groups. Furthermore, there was excess mortality one to six months after infection with Yersinia enterocolitica (relative risk 2.53, 95% confidence interval 1.38 to 4.62) and from six months to one year after infection with Campylobacter (1.35, 1.02 to 1.80) and Salmonella (1.53, 1.31 to 1.79). CONCLUSIONS: Infections with all these bacteria were associated with an increased short term risk of death, even after pre-existing illnesses were taken into account. Salmonella, Campylobacter, and Yersinia enterocolitica infections were also associated with increased long term mortality. PMID- 12586667 TI - Prognostic indicators of early and late death in children admitted to district hospital in Kenya: cohort study. AB - OBJECTIVES: To identify clinical indicators of immediate, early, and late mortality in children at admission to a sub-Saharan district hospital and to develop prognostic scores. DESIGN: Prospective cohort study. SETTING: One district hospital in Kenya. PARTICIPANTS: Children aged over 90 days admitted to hospital from 1 July 1998 to 30 June 2001. MAIN OUTCOME MEASURES: Prognostic indicators of mortality. RESULTS: Of 8091 children admitted up to 1 June 2000, 436 (5%) died. Sixty (14%) died within four hours after admission (immediate), 193 (44%) after 4-48 hours (early), and 183 (42%) after 48 hours (late). There were marked differences in the clinical features associated with immediate, early, and late death. Seven indicators (neurological status, respiratory distress (subcostal indrawing or deep breathing), nutritional status (wasting or kwashiorkor), severe anaemia, jaundice, axillary temperature, and length of history) were included in simplified prognostic scores. Data from 4802 children admitted from 1 July 2000 to 30 June 2001 were used to validate the scores. For simplified prognostic scores the areas under the receiver operating characteristic curves were 0.93 (95% confidence interval 0.92 to 0.94), 0.82 (0.80 to 0.83), and 0.82 (0.81 to 0.84) for immediate, early, and late death, respectively. CONCLUSION: In children admitted to a sub-Saharan hospital, the prognostic indicators of early and late deaths differ but a small number of simple clinical signs predict outcome well. PMID- 12586668 TI - Cross sectional survey of meningococcal C immunisation in children looked after by local authorities and those living at home. PMID- 12586669 TI - Herd immunity from meningococcal serogroup C conjugate vaccination in England: database analysis. PMID- 12586670 TI - Effect of public education aimed at early diagnosis of malignant melanoma: cohort comparison study. PMID- 12586671 TI - Dying from cancer in developed and developing countries: lessons from two qualitative interview studies of patients and their carers. AB - OBJECTIVE: To describe the experiences of illness and needs and use of services in two groups of patients with incurable cancer, one in a developed country and the other in a developing country. DESIGN: Scotland: longitudinal study with qualitative interviews. Kenya: cross sectional study with qualitative interviews. SETTINGS: Lothian region, Scotland, and Meru District, Kenya. PARTICIPANTS: Scotland: 20 patients with inoperable lung cancer and their carers. Kenya: 24 patients with common advanced cancers and their main informal carers. MAIN OUTCOME MEASURES: Descriptions of experiences, needs, and available services. RESULTS: 67 interviews were conducted in Scotland and 46 in Kenya. The emotional pain of facing death was the prime concern of Scottish patients and their carers, while physical pain and financial worries dominated the lives of Kenyan patients and their carers. In Scotland, free health and social services (including financial assistance) were available, but sometimes underused. In Kenya, analgesia, essential equipment, suitable food, and assistance in care were often inaccessible and unaffordable, resulting in considerable unmet physical needs. Kenyan patients thought that their psychological, social, and spiritual needs were met by their families, local community, and religious groups. Some Scottish patients thought that such non-physical needs went unmet. CONCLUSIONS: In patients living in developed and developing countries there are differences not only in resources available for patients dying from cancer but also in their lived experience of illness. The expression of needs and how they are met in different cultural contexts can inform local assessment of needs and provide insights for initiatives in holistic cancer care. PMID- 12586672 TI - Association between practice size and quality of care of patients with ischaemic heart disease: cross sectional study. PMID- 12586673 TI - Patients' consent preferences for research uses of information in electronic medical records: interview and survey data. AB - OBJECTIVES: To assess patients' preferred method of consent for the use of information from electronic medical records for research. DESIGN: Interviews and a structured survey of patients in practices with electronic medical records. SETTING: Family practices in southern Ontario, Canada. PARTICIPANTS: 123 patients: 17 were interviewed and 106 completed a survey. MAIN OUTCOME MEASURES: Patients' opinions and concerns on use of information from their medical records for research and their preferences for method of consent. RESULTS: Most interviewees were willing to allow the use of their information for research purposes, although the majority preferred that consent was sought first. The seeking of consent was considered an important element of respect for the individual. Most interviewees made little distinction between identifiable and anonymised data. Research sponsored by private insurance firms generated the greatest concern, and research sponsored by foundation the least. Sponsorship by drug companies evoked negative responses during interview and positive responses in the survey. CONCLUSIONS: Patients are willing to allow information from their medical records to be used for research, but most prefer to be asked for consent either verbally or in writing. PMID- 12586675 TI - Fatal dysnatraemia caused by elective colonoscopy. PMID- 12586674 TI - Leishmaniasis: new approaches to disease control. PMID- 12586676 TI - ABC of learning and teaching in medicine: Evaluation. PMID- 12586677 TI - Problems with UK government's risk sharing scheme for assessing drugs for multiple sclerosis. PMID- 12586678 TI - Indiscriminate investigations have adverse effects. PMID- 12586679 TI - Simple presentation of test accuracy may lead to inflated disease probabilities. PMID- 12586680 TI - Effect of computerised evidence based guidelines. Computer support is complex intervention. PMID- 12586681 TI - Novartis was not in breach of code for "inventing" disease. PMID- 12586682 TI - Assortative mating may explain spouses' risk of same disease. PMID- 12586683 TI - Drug misuse should always be considered in young people with impaired consciousness. PMID- 12586684 TI - Unit of analysis errors should be clarified in meta-analyses. PMID- 12586685 TI - GPs can separate oncological wheat from chaff. PMID- 12586686 TI - Look before you flush. PMID- 12586691 TI - One hundred years of mouse genetics: an intellectual history. I. The classical period (1902-1980). PMID- 12586692 TI - Genetic analysis of the interface between Cdc42p and the CRIB domain of Ste20p in Saccharomyces cerevisiae. AB - Mutagenesis was used to probe the interface between the small GTPase Cdc42p and the CRIB domain motif of Ste20p. Members of a cluster of hydrophobic residues of Cdc42p were changed to alanine and/or arginine. The interaction of the wild-type and mutant proteins was measured using the two-hybrid assay; many, but not all, changes reduced interaction between Cdc42p and the target CRIB domain. Mutations in conserved residues in the CRIB domain were also tested for their importance in the association with Cdc42p. Two conserved CRIB domain histidines were changed to aspartic acid. These mutants reduced mating, as well as responsiveness to pheromone-induced gene expression and cell cycle arrest, but did not reduce in vitro the kinase activity of Ste20p. GFP-tagged mutant proteins were unable to localize to sites of polarized growth. In addition, these point mutants were synthetically lethal with disruption of CLA4 and blocked the Ste20p-Cdc42p two hybrid interaction. Compensatory mutations in Cdc42p that reestablished the two hybrid association with the mutant Ste20p CRIB domain baits were identified. These mutations improved the pheromone responsiveness of cells containing the CRIB mutations, but did not rescue the lethality associated with the CRIB mutant CLA4 deletion interaction. These results suggest that the Ste20p-Cdc42p interaction plays a direct role in Ste20p kinase function and that this interaction is required for efficient activity of the pheromone response pathway. PMID- 12586693 TI - Loss of CDC5 function in Saccharomyces cerevisiae leads to defects in Swe1p regulation and Bfa1p/Bub2p-independent cytokinesis. AB - In many organisms, polo kinases appear to play multiple roles during M-phase progression. To provide new insights into the function of budding yeast polo kinase Cdc5p, we generated novel temperature-sensitive cdc5 mutants by mutagenizing the C-terminal domain. Here we show that, at a semipermissive temperature, the cdc5-3 mutant exhibited a synergistic bud elongation and growth defect with loss of HSL1, a component important for normal G(2)/M transition. Loss of SWE1, which phosphorylates and inactivates the budding yeast Cdk1 homolog Cdc28p, suppressed the cdc5-3 hsl1Delta defect, suggesting that Cdc5p functions at a point upstream of Swe1p. In addition, the cdc5-4 and cdc5-7 mutants exhibited chained cell morphologies with shared cytoplasms between the connected cell bodies, indicating a cytokinetic defect. Close examination of these mutants revealed delayed septin assembly at the incipient bud site and loosely organized septin rings at the mother-bud neck. Components in the mitotic exit network (MEN) play important roles in normal cytokinesis. However, loss of BFA1 or BUB2, negative regulators of the MEN, failed to remedy the cytokinetic defect of these mutants, indicating that Cdc5p promotes cytokinesis independently of Bfa1p and Bub2p. Thus, Cdc5p contributes to the activation of the Swe1p-dependent Cdc28p/Clb pathway, normal septin function, and cytokinesis. PMID- 12586694 TI - SOD2 functions downstream of Sch9 to extend longevity in yeast. AB - Signal transduction pathways inactivated during periods of starvation are implicated in the regulation of longevity in organisms ranging from yeast to mammals, but the mechanisms responsible for life-span extension are poorly understood. Chronological life-span extension in S. cerevisiae cyr1 and sch9 mutants is mediated by the stress-resistance proteins Msn2/Msn4 and Rim15. Here we show that mitochondrial superoxide dismutase (Sod2) is required for survival extension in yeast. Deletion of SOD2 abolishes life-span extension in sch9Delta mutants and decreases survival in cyr1:mTn mutants. The overexpression of Sods- mitochondrial Sod2 and cytosolic CuZnSod (Sod1)--delays the age-dependent reversible inactivation of mitochondrial aconitase, a superoxide-sensitive enzyme, and extends survival by 30%. Deletion of the RAS2 gene, which functions upstream of CYR1, also doubles the mean life span by a mechanism that requires Msn2/4 and Sod2. These findings link mutations that extend chronological life span in S. cerevisiae to superoxide dismutases and suggest that the induction of other stress-resistance genes regulated by Msn2/4 and Rim15 is required for maximum longevity extension. PMID- 12586695 TI - Large-scale functional genomic analysis of sporulation and meiosis in Saccharomyces cerevisiae. AB - We have used a single-gene deletion mutant bank to identify the genes required for meiosis and sporulation among 4323 nonessential Saccharomyces cerevisiae annotated open reading frames (ORFs). Three hundred thirty-four sporulation essential genes were identified, including 78 novel ORFs and 115 known genes without previously described sporulation defects in the comprehensive Saccharomyces Genome (SGD) or Yeast Proteome (YPD) phenotype databases. We have further divided the uncharacterized sporulation-essential genes into early, middle, and late stages of meiosis according to their requirement for IME1 induction and nuclear division. We believe this represents a nearly complete identification of the genes uniquely required for this complex cellular pathway. The set of genes identified in this phenotypic screen shows only limited overlap with those identified by expression-based studies. PMID- 12586696 TI - The Rad27 (Fen-1) nuclease inhibits Ty1 mobility in Saccharomyces cerevisiae. AB - Although most Ty1 elements in Saccharomyces cerevisiae are competent for retrotransposition, host defense genes can inhibit different steps of the Ty1 life cycle. Here, we demonstrate that Rad27, a structure-specific nuclease that plays an important role in DNA replication and genome stability, inhibits Ty1 at a post-translational level. We have examined the effects of various rad27 mutations on Ty1 element retrotransposition and cDNA recombination, termed Ty1 mobility. The point mutations rad27-G67S, rad27-G240D, and rad27-E158D that cause defects in certain enzymatic activities in vitro result in variable increases in Ty1 mobility, ranging from 4- to 22-fold. The C-terminal frameshift mutation rad27-324 confers the maximum increase in Ty1 mobility (198-fold), unincorporated cDNA, and insertion at preferred target sites. The null mutation differs from the other rad27 alleles by increasing the frequency of multimeric Ty1 insertions and cDNA recombination with a genomic element. The rad27 mutants do not markedly alter the levels of Ty1 RNA or the TyA1-gag protein. However, there is an increase in the stability of unincorporated Ty1 cDNA in rad27-324 and the null mutant. Our results suggest that Rad27 inhibits Ty1 mobility by destabilizing unincorporated Ty1 cDNA and preventing the formation of Ty1 multimers. PMID- 12586697 TI - Specialization of function among aldehyde dehydrogenases: the ALD2 and ALD3 genes are required for beta-alanine biosynthesis in Saccharomyces cerevisiae. AB - The amino acid beta-alanine is an intermediate in pantothenic acid (vitamin B(5)) and coenzyme A (CoA) biosynthesis. In contrast to bacteria, yeast derive the beta alanine required for pantothenic acid production via polyamine metabolism, mediated by the four SPE genes and by the FAD-dependent amine oxidase encoded by FMS1. Because amine oxidases generally produce aldehyde derivatives of amine compounds, we propose that an additional aldehyde-dehydrogenase-mediated step is required to make beta-alanine from the precursor aldehyde, 3-aminopropanal. This study presents evidence that the closely related aldehyde dehydrogenase genes ALD2 and ALD3 are required for pantothenic acid biosynthesis via conversion of 3 aminopropanal to beta-alanine in vivo. While deletion of the nuclear gene encoding the unrelated mitochondrial Ald5p resulted in an enhanced requirement for pantothenic acid pathway metabolites, we found no evidence to indicate that the Ald5p functions directly in the conversion of 3-aminopropanal to beta alanine. Thus, in Saccharomyces cerevisiae, ALD2 and ALD3 are specialized for beta-alanine biosynthesis and are consequently involved in the cellular biosynthesis of coenzyme A. PMID- 12586698 TI - Genetic diversity in yeast assessed with whole-genome oligonucleotide arrays. AB - The availability of a complete genome sequence allows the detailed study of intraspecies variability. Here we use high-density oligonucleotide arrays to discover 11,115 single-feature polymorphisms (SFPs) existing in one or more of 14 different yeast strains. We use these SFPs to define regions of genetic identity between common laboratory strains of yeast. We assess the genome-wide distribution of genetic variation on the basis of this yeast population. We find that genome variability is biased toward the ends of chromosomes and is more likely to be found in genes with roles in fermentation or in transport. This subtelomeric bias may arise through recombination between nonhomologous sequences because full-gene deletions are more common in these regions than in more central regions of the chromosome. PMID- 12586699 TI - Investigation of the stability of yeast rad52 mutant proteins uncovers post translational and transcriptional regulation of Rad52p. AB - We investigated the stability of the Saccharomyces cerevisiae Rad52 protein to learn how a cell controls its quantity and longevity. We measured the cellular levels of wild-type and mutant forms of Rad52p when expressed from the RAD52 promoter and the half-lives of the various forms of Rad52p when expressed from the GAL1 promoter. The wild-type protein has a half-life of 15 min. rad52 mutations variably affect the cellular levels of the protein products, and these levels correlate with the measured half-lives. While missense mutations in the N terminus of the protein drastically reduce the cellular levels of the mutant proteins, two mutations--one a deletion of amino acids 210-327 and the other a missense mutation of residue 235--increase the cellular level and half-life more than twofold. These results suggest that Rad52p is subject to post-translational regulation. Proteasomal mutations have no effect on Rad52p half-life but increase the amount of RAD52 message. In contrast to Rad52p, the half-life of Rad51p is >2 hr, and RAD51 expression is unaffected by proteasomal mutations. These differences between Rad52p and Rad51p suggest differential regulation of two proteins that interact in recombinational repair. PMID- 12586700 TI - Roles for WHITE COLLAR-1 in circadian and general photoperception in Neurospora crassa. AB - The transcription factors WHITE COLLAR-1 (WC-1) and WHITE COLLAR-2 (WC-2) interact to form a heterodimeric complex (WCC) that is essential for most of the light-mediated processes in Neurospora crassa. WCC also plays a distinct non light-related role as the transcriptional activator in the FREQUENCY (FRQ)/WCC feedback loop that is central to the N. crassa circadian system. Although an activator role was expected for WC-1, unanticipated phenotypes resulting from some wc-1 alleles prompted a closer examination of an allelic series for WC-1 that has uncovered roles for this central regulator in constant darkness and in response to light. We analyzed the phenotypes of five different wc-1 mutants for expression of FRQ and WC-1 in constant darkness and following light induction. While confirming the absolute requirement of WC-1 for light responses, the data suggest multiple levels of control for light-regulated genes. PMID- 12586701 TI - Genetic analysis of Caenorhabditis elegans glp-1 mutants suggests receptor interaction or competition. AB - glp-1 encodes a member of the highly conserved LIN-12/Notch family of receptors that mediates the mitosis/meiosis decision in the C. elegans germline. We have characterized three mutations that represent a new genetic and phenotypic class of glp-1 mutants, glp-1(Pro). The glp-1(Pro) mutants display gain-of-function germline pattern defects, most notably a proximal proliferation (Pro) phenotype. Each of three glp-1(Pro) alleles encodes a single amino acid change in the extracellular part of the receptor: two in the LIN-12/Notch repeats (LNRs) and one between the LNRs and the transmembrane domain. Unlike other previously described gain-of-function mutations that affect this region of LIN-12/Notch family receptors, the genetic behavior of glp-1(Pro) alleles is not consistent with simple hypermorphic activity. Instead, the mutant phenotype is suppressed by wild-type doses of glp-1. Moreover, a trans-heterozygous combination of two highly penetrant glp-1(Pro) mutations is mutually suppressing. These results lend support to a model for a higher-order receptor complex and/or competition among receptor proteins for limiting factors that are required for proper regulation of receptor activity. Double-mutant analysis with suppressors and enhancers of lin 12 and glp-1 further suggests that the functional defect in glp-1(Pro) mutants occurs prior to or at the level of ligand interaction. PMID- 12586702 TI - Control of vulval competence and centering in the nematode Oscheius sp. 1 CEW1. AB - To compare vulva development mechanisms in the nematode Oscheius sp. 1 to those known in Caenorhabditis elegans, we performed a genetic screen for vulva mutants in Oscheius sp. 1 CEW1. Here we present one large category of mutations that we call cov, which affect the specification of the Pn.p ventral epidermal cells along the antero-posterior axis. The Pn.p cells are numbered from 1 to 12 from anterior to posterior. In wild-type Oscheius sp. 1 CEW1, the P(4-8).p cells are competent to form the vulva and the progeny of P(5-7).p actually form the vulva, with the descendants of P6.p adopting a central vulval fate. Among the 17 mutations (defining 13 genes) that we characterize here, group 1 mutations completely or partially abolish P(4-8).p competence, and this correlates with early fusion of the Pn.p cells to the epidermal syncytium. In this group, we found a putative null mutation in the lin-39 HOM-C homolog, the associated phenotype of which could be weakly mimicked by injection of a morpholino against Osp1-lin-39 in the mother's germ line. Using cell ablation in a partially penetrant competence mutant, we show that vulval competence is partially controlled by a gonadal signal. Most other mutants found in the screen display phenotypes unknown in C. elegans. Group 2 mutants show a partial penetrance of Pn.p competence loss and an abnormal centering of the vulva on P5.p, suggesting that these two processes are coregulated by the same pathway in Oscheius sp. 1. Group 3 mutants display an enlarged competence group that includes P3.p, thus demonstrating the existence of a specific mechanism inhibiting P3.p competence. Group 4 mutants display an abnormal centering of the vulval pattern on P7.p and suggest that a specific mechanism centers the vulval pattern on a single Pn.p cell. PMID- 12586703 TI - Population genetics of Caenorhabditis elegans: the paradox of low polymorphism in a widespread species. AB - Caenorhabditis elegans has become one of the most widely used model research organisms, yet we have little information on evolutionary processes and recent evolutionary history of this widespread species. We examined patterns of variation at 20 microsatellite loci in a sample of 23 natural isolates of C. elegans from various parts of the world. One-half of the loci were monomorphic among all strains, and overall genetic variation at microsatellite loci was low, relative to most other species. Some population structure was detected, but there was no association between the genetic and geographic distances among different natural isolates. Thus, despite the nearly worldwide occurrence of C. elegans, little evidence was found for local adaptation in strains derived from different parts of the world. The low levels of genetic variation within and among populations suggest that recent colonization and population expansion might have occurred. However, the patterns of variation are not consistent with population expansion. A possible explanation for the observed patterns is the action of background selection to reduce polymorphism, coupled with ongoing gene flow among populations worldwide. PMID- 12586704 TI - Suppression of the ELO-2 FA elongation activity results in alterations of the fatty acid composition and multiple physiological defects, including abnormal ultradian rhythms, in Caenorhabditis elegans. AB - While the general steps of fatty acid (FA) biosynthesis are well understood, the individual enzymes involved in the elongation of long chain saturated and polyunsaturated FA (PUFA) are largely unknown. Recent research indicates that these enzymes might be of considerable physiological importance for human health. We use Caenorhabditis elegans to study FA elongation activities and associated abnormal phenotypes. In this article we report that the predicted C. elegans F11E6.5/ELO-2 is a functional enzyme with the FA elongation activity. It is responsible for the elongation of palmitic acid and is involved in PUFA biosynthesis. RNAi-mediated suppression of ELO-2 causes an accumulation of palmitate and an associated decrease in the PUFA fraction in triacylglycerides and phospholipid classes. This imbalance in the FA composition results in multiple phenotypic defects such as slow growth, small body size, reproductive defects, and changes in rhythmic behavior. ELO-2 cooperates with the previously reported ELO-1 in 20-carbon PUFA production, and at least one of the enzymes must function to provide normal growth and development in C. elegans. The presented data indicate that suppression of a single enzyme of the FA elongation machinery is enough to affect various organs and systems in worms. This effect resembles syndromic disorders in humans. PMID- 12586705 TI - Positive selection of Caenorhabditis elegans mutants with increased stress resistance and longevity. AB - We developed selective conditions for long-lived mutants of the nematode Caenorhabditis elegans by subjecting the first larval stage (L1) to thermal stress at 30 degrees for 7 days. The surviving larvae developed to fertile adults after the temperature was shifted to 15 degrees. A total of one million F(2) progeny and a half million F(3) progeny of ethyl-methanesulfonate-mutagenized animals were treated in three separate experiments. Among the 81 putative mutants that recovered and matured to the reproductive adult, 63 retested as thermotolerant and 49 (80%) exhibited a >15% increase in mean life span. All the known classes of dauer formation (Daf) mutant that affect longevity were found, including six new alleles of daf-2, and a unique temperature-sensitive, dauer constitutive allele of age-1. Alleles of dyf-2 and unc-13 were isolated, and mutants of unc-18, a gene that interacts with unc-13, were also found to be long lived. Thirteen additional mutations define at least four new genes. PMID- 12586706 TI - Sequence variation of alcohol dehydrogenase (Adh) paralogs in cactophilic Drosophila. AB - This study focuses on the population genetics of alcohol dehydrogenase (Adh) in cactophilic Drosophila. Drosophila mojavensis and D. arizonae utilize cactus hosts, and each host contains a characteristic mixture of alcohol compounds. In these Drosophila species there are two functional Adh loci, an adult form (Adh-2) and a larval and ovarian form (Adh-1). Overall, the greater level of variation segregating in D. arizonae than in D. mojavensis suggests a larger population size for D. arizonae. There are markedly different patterns of variation between the paralogs across both species. A 16-bp intron haplotype segregates in both species at Adh-2, apparently the product of an ancient gene conversion event between the paralogs, which suggests that there is selection for the maintenance of the intron structure possibly for the maintenance of pre-mRNA structure. We observe a pattern of variation consistent with adaptive protein evolution in the D. mojavensis lineage at Adh-1, suggesting that the cactus host shift that occurred in the divergence of D. mojavensis from D. arizonae had an effect on the evolution of the larval expressed paralog. Contrary to previous work we estimate a recent time for both the divergence of D. mojavensis and D. arizonae (2.4 +/- 0.7 MY) and the age of the gene duplication (3.95 +/- 0.45 MY). PMID- 12586707 TI - A P-element insertion screen identified mutations in 455 novel essential genes in Drosophila. AB - With the completion of the nucleotide sequences of several complex eukaryotic genomes, tens of thousands of genes have been predicted. However, this information has to be correlated with the functions of those genes to enhance our understanding of biology and to improve human health care. The Drosophila transposon P-element-induced mutations are very useful for directly connecting gene products to their biological function. We designed an efficient transposon P element-mediated gene disruption procedure and performed genetic screening for single P-element insertion mutations, enabling us to recover 2500 lethal mutations. Among these, 2355 are second chromosome mutations. Sequences flanking >2300 insertions that identify 850 different genes or ESTs (783 genes on the second chromosome and 67 genes on the third chromosome) have been determined. Among these, 455 correspond to genes for which no lethal mutation has yet been reported. The Drosophila genome is thought to contain approximately 3600 vital genes; 1400 are localized on the second chromosome. Our mutation collection represents approximately 56% of the second chromosome vital genes and approximately 24% of the total vital Drosophila genes. PMID- 12586708 TI - Analysis of Ras-induced overproliferation in Drosophila hemocytes. AB - We use the Drosophila melanogaster larval hematopoietic system as an in vivo model for the genetic and functional genomic analysis of oncogenic cell overproliferation. Ras regulates cell proliferation and differentiation in multicellular eukaryotes. To further elucidate the role of activated Ras in cell overproliferation, we generated a collagen promoter-Gal4 strain to overexpress Ras(V12) in Drosophila hemocytes. Activated Ras causes a dramatic increase in the number of circulating larval hemocytes (blood cells), which is caused by cellular overproliferation. This phenotype is mediated by the Raf/MAPK pathway. The mutant hemocytes retain the ability to phagocytose bacteria as well as to differentiate into lamellocytes. Microarray analysis of hemocytes overexpressing Ras(V12) vs. Ras(+) identified 279 transcripts that are differentially expressed threefold or more in hemocytes expressing activated Ras. This work demonstrates that it will be feasible to combine genetic and functional genomic approaches in the Drosophila hematopoietic system to systematically identify oncogene-specific downstream targets. PMID- 12586709 TI - A novel system of fertility rescue in Drosophila hybrids reveals a link between hybrid lethality and female sterility. AB - Hybrid daughters of crosses between Drosophila melanogaster females and males from the D. simulans species clade are fully viable at low temperature but have agametic ovaries and are thus sterile. We report here that mutations in the D. melanogaster gene Hybrid male rescue (Hmr), along with unidentified polymorphic factors, rescue this agametic phenotype in both D. melanogaster/D. simulans and D. melanogaster/D. mauritiana F(1) female hybrids. These hybrids produced small numbers of progeny in backcrosses, their low fecundity being caused by incomplete rescue of oogenesis as well as by zygotic lethality. F(1) hybrid males from these crosses remained fully sterile. Hmr(+) is the first Drosophila gene shown to cause hybrid female sterility. These results also suggest that, while there is some common genetic basis to hybrid lethality and female sterility in D. melanogaster, hybrid females are more sensitive to fertility defects than to lethality. PMID- 12586710 TI - amontillado, the Drosophila homolog of the prohormone processing protease PC2, is required during embryogenesis and early larval development. AB - Biosynthesis of most peptide hormones and neuropeptides requires proteolytic excision of the active peptide from inactive proprotein precursors, an activity carried out by subtilisin-like proprotein convertases (SPCs) in constitutive or regulated secretory pathways. The Drosophila amontillado (amon) gene encodes a homolog of the mammalian PC2 protein, an SPC that functions in the regulated secretory pathway in neuroendocrine tissues. We have identified amon mutants by isolating ethylmethanesulfonate (EMS)-induced lethal and visible mutations that define two complementation groups in the amon interval at 97D1 of the third chromosome. DNA sequencing identified the amon complementation group and the DNA sequence change for each of the nine amon alleles isolated. amon mutants display partial embryonic lethality, are defective in larval growth, and arrest during the first to second instar larval molt. Mutant larvae can be rescued by heat shock-induced expression of the amon protein. Rescued larvae arrest at the subsequent larval molt, suggesting that amon is also required for the second to third instar larval molt. Our data indicate that the amon proprotein convertase is required during embryogenesis and larval development in Drosophila and support the hypothesis that AMON acts to proteolytically process peptide hormones that regulate hatching, larval growth, and larval ecdysis. PMID- 12586711 TI - In vivo introduction of unpreferred synonymous codons into the Drosophila Adh gene results in reduced levels of ADH protein. AB - The evolution of codon bias, the unequal usage of synonymous codons, is thought to be due to natural selection for the use of preferred codons that match the most abundant species of isoaccepting tRNA, resulting in increased translational efficiency and accuracy. We examined this hypothesis by introducing 1, 6, and 10 unpreferred codons into the Drosophila alcohol dehydrogenase gene (Adh). We observed a significant decrease in ADH protein production with number of unpreferred codons, confirming the importance of natural selection as a mechanism leading to codon bias. We then used this empirical relationship to estimate the selection coefficient (s) against unpreferred synonymous mutations and found the value (s >or= 10(-5)) to be approximately one order of magnitude greater than previous estimates from population genetics theory. The observed differences in protein production appear to be too large to be consistent with current estimates of the strength of selection on synonymous sites in D. melanogaster. PMID- 12586712 TI - Common spontaneous sex-reversed XX males of the medaka Oryzias latipes. AB - In the medaka, a duplicated version of the dmrt1 gene, dmrt1bY, has been identified as a candidate for the master male sex-determining gene on the Y chromosome. By screening several strains of Northern and Southern medaka we identified a considerable number of males with normal phenotype and uncompromised fertility, but lacking dmrt1bY. The frequency of such males was >10% in some strains and zero in others. Analysis for the presence of other Y-linked markers by FISH analysis, PCR, and phenotype indicated that their genotype is XX. Crossing such males with XX females led to a strong female bias in the offspring and also to a reappearance of XX males in the following generations. This indicated that the candidate male sex-determining gene dmrt1bY may not be necessary for male development in every case, but that its function can be taken over by so far unidentified autosomal modifiers. PMID- 12586713 TI - Molecular dissection of a quantitative trait locus: a phenylalanine-to-tyrosine substitution in the transmembrane domain of the bovine growth hormone receptor is associated with a major effect on milk yield and composition. AB - We herein report on our efforts to improve the mapping resolution of a QTL with major effect on milk yield and composition that was previously mapped to bovine chromosome 20. By using a denser chromosome 20 marker map and by exploiting linkage disequilibrium using two distinct approaches, we provide strong evidence that a chromosome segment including the gene coding for the growth hormone receptor accounts for at least part of the chromosome 20 QTL effect. By sequencing individuals with known QTL genotype, we identify an F to Y substitution in the transmembrane domain of the growth hormone receptor gene that is associated with a strong effect on milk yield and composition in the general population. PMID- 12586714 TI - Interallelic complementation at the mouse Mitf locus. AB - Mutations at the mouse microphthalmia locus (Mitf) affect the development of different cell types, including melanocytes, retinal pigment epithelial cells of the eye, and osteoclasts. The MITF protein is a member of the MYC supergene family of basic-helix-loop-helix-leucine-zipper (bHLHZip) transcription factors and is known to regulate the expression of cell-specific target genes by binding DNA as homodimer or as heterodimer with related proteins. The many mutations isolated at the locus have different effects on the phenotype and can be arranged in an allelic series in which the phenotypes range from near normal to white microphthalmic animals with osteopetrosis. Previous investigations have shown that certain combinations of Mitf alleles complement each other, resulting in a phenotype more normal than that of each homozygote alone. Here we analyze this interallelic complementation in detail and show that it is limited to one particular allele, Mitf(Mi-white) (Mitf(Mi-wh)), a mutation affecting the DNA binding domain. Both loss- and gain-of-function mutations are complemented, as are other Mitf mutations affecting the DNA-binding domain. Furthermore, this behavior is not restricted to particular cell types: Both eye development and coat color phenotypes are complemented. Our analysis suggests that Mitf(Mi-wh) associated interallelic complementation is due to the unique biochemical nature of this mutation. PMID- 12586715 TI - Effects of genetic background on response to selection in experimental populations of Arabidopsis thaliana. AB - The extent to which genetic background can influence allelic fitness is poorly understood, despite having important evolutionary consequences. Using experimental populations of Arabidopsis thaliana and map-based population genetic data, we examined a multigeneration response to selection in populations with differentiated genetic backgrounds. Replicated experimental populations of A. thaliana with genetic backgrounds derived from ecotypes Landsberg and Niederzenz were subjected to strong viability and fertility selection by growing individuals from each population at high density for three generations in a growth chamber. Patterns of genome-wide selection were evaluated by examining deviations from expected frequencies of mapped molecular markers. Estimates of selection coefficients for individual genomic regions ranged from near 0 to 0.685. Genomic regions demonstrating the strongest response to selection most often were selected similarly in both genetic backgrounds. The selection response of several weakly selected regions, however, appeared to be sensitive to genetic background, but only one region showed evidence of positive selection in one background and negative selection in another. These results are most consistent with models of adaptive evolution in which allelic fitnesses are not strongly influenced by genetic background and only infrequently change in sign due to variation at other loci. PMID- 12586716 TI - The role of 2n gametes and endosperm balance number in the origin and evolution of polyploids in the tuber-bearing Solanums. AB - Polyploidization has played a major role in the origin and evolution of polyploid species. In this article we outline the unique characteristics of 2n gametes and implications of their participation in the evolution of polyploid Solanum species. The genetic consequences of 2n gametes indicate that sexual polyploidization results in greater variability, fitness, and heterozygosity than does somatic doubling. Further, the mechanisms of 2n gamete formation and the frequency of 2n gamete-forming genes in present polyploids and their ancestral species provide additional evidence of their involvement. Equally important is the endosperm, via the endosperm balance number (EBN) incompatibility system, in complementing the role of 2n gametes. In fact, the EBN system acts as a screen for either 1n or 2n gametes, depending on the EBN and chromosome numbers of parental species. EBN in combination with 2n gametes maintains the ploidy integrity of diploid ancestral species, while providing the flexibility for either unilateral or bilateral sexual polyploidization. PMID- 12586717 TI - Cloning of the pleiotropic T locus in soybean and two recessive alleles that differentially affect structure and expression of the encoded flavonoid 3' hydroxylase. AB - Three loci (I, R, and T) control pigmentation of the seed coats in Glycine max and are genetically distinct from those controlling flower color. The T locus also controls color of the trichome hairs. We report the identification and isolation of a flavonoid 3' hydroxylase gene from G. max (GmF3'H) and the linkage of this gene to the T locus. This GmF3'H gene was highly expressed in early stages of seed coat development and was expressed at very low levels or not at all in other tissues. Evidence that the GmF3'H gene is linked to the T locus came from the occurrence of multiple RFLPs in lines with varying alleles of the T locus, as well as in a population of plants segregating at that locus. GmF3'H genomic and cDNA sequence analysis of color mutant lines with varying t alleles revealed a frameshift mutation in one of the alleles. In another line derived from a mutable genetic stock, the abundance of the mRNAs for GmF3'H was dramatically reduced. Isolation of the GmF3'H gene and its identification as the T locus will enable investigation of the pleiotropic effects of the T locus on cell wall integrity and its involvement in the regulation of the multiple branches of the flavonoid pathway in soybean. PMID- 12586718 TI - Unexpected complexity of poly(A)-binding protein gene families in flowering plants: three conserved lineages that are at least 200 million years old and possible auto- and cross-regulation. AB - Eukaryotic poly(A)-binding protein (PABP) is a ubiquitous, essential factor involved in mRNA biogenesis, translation, and turnover. Most eukaryotes examined have only one or a few PABPs. In contrast, eight expressed PABP genes are present in Arabidopsis thaliana. These genes fall into three distinct classes, based on highly concordant results of (i) phylogenetic analysis of the amino acid sequences of the encoded proteins, (ii) analysis of the intron number and placement, and (iii) surveys of gene expression patterns. Representatives of each of the three classes also exist in the rice genome, suggesting that the diversification of the plant PABP genes has occurred prior to the split of monocots and dicots >or=200 MYA. Experiments with the recombinant PAB3 protein suggest the possibility of a negative feedback regulation, as well as of cross regulation between the Arabidopsis PABPs that belong to different classes but are simultaneously expressed in the same cell type. Such a high complexity of the plant PABPs might enable a very fine regulation of organismal growth and development at the post-transcriptional level, compared with PABPs of other eukaryotes. PMID- 12586719 TI - Genetic and functional analysis of DD44, a sex-linked gene from the dioecious plant Silene latifolia, provides clues to early events in sex chromosome evolution. AB - Silene latifolia is a dioecious plant with heteromorphic sex chromosomes. The sex chromosomes of S. latifolia provide an opportunity to study the early events in sex chromosome evolution because of their relatively recent emergence. In this article, we present the genetic and physical mapping, expression analysis, and molecular evolutionary analysis of a sex-linked gene from S. latifolia, DD44 (Differential Display 44). DD44 is homologous to the oligomycin sensitivity conferring protein, an essential component of the mitochondrial ATP synthase, and is ubiquitously expressed in both sexes. We have been able to genetically map DD44 to a region of the Y chromosome that is genetically linked to the carpel suppressing locus. Although we have physically mapped DD44 to the distal end of the long arm of the X chromosome using fluorescence in situ hybridization (FISH), DD44 maps to the opposite arm of the Y chromosome as determined by our genetic map. These data suggest that chromosomal rearrangements have occurred on the Y chromosome, which may have contributed to the genetic isolation of the Y chromosome. We discuss the implications of these results with respect to the structural and functional evolution of the S. latifolia Y chromosome. PMID- 12586720 TI - The inheritance of chemical phenotype in Cannabis sativa L. AB - Four crosses were made between inbred Cannabis sativa plants with pure cannabidiol (CBD) and pure Delta-9-tetrahydrocannabinol (THC) chemotypes. All the plants belonging to the F(1)'s were analyzed by gas chromatography for cannabinoid composition and constantly found to have a mixed CBD-THC chemotype. Ten individual F(1) plants were self-fertilized, and 10 inbred F(2) offspring were collected and analyzed. In all cases, a segregation of the three chemotypes (pure CBD, mixed CBD-THC, and pure THC) fitting a 1:2:1 proportion was observed. The CBD/THC ratio was found to be significantly progeny specific and transmitted from each F(1) to the F(2)'s derived from it. A model involving one locus, B, with two alleles, B(D) and B(T), is proposed, with the two alleles being codominant. The mixed chemotypes are interpreted as due to the genotype B(D)/B(T) at the B locus, while the pure-chemotype plants are due to homozygosity at the B locus (either B(D)/B(D) or B(T)/B(T)). It is suggested that such codominance is due to the codification by the two alleles for different isoforms of the same synthase, having different specificity for the conversion of the common precursor cannabigerol into CBD or THC, respectively. The F(2) segregating groups were used in a bulk segregant analysis of the pooled DNAs for screening RAPD primers; three chemotype-associated markers are described, one of which has been transformed in a sequence-characterized amplified region (SCAR) marker and shows tight linkage to the chemotype and codominance. PMID- 12586721 TI - On marker-assisted prediction of genetic value: beyond the ridge. AB - Marked-assisted genetic improvement of agricultural species exploits statistical dependencies in the joint distribution of marker genotypes and quantitative traits. An issue is how molecular (e.g., dense marker maps) and phenotypic information (e.g., some measure of yield in plants) is to be used for predicting the genetic value of candidates for selection. Multiple regression, selection index techniques, best linear unbiased prediction, and ridge regression of phenotypes on marker genotypes have been suggested, as well as more elaborate methods. Here, phenotype-marker associations are modeled hierarchically via multilevel models including chromosomal effects, a spatial covariance of marked effects within chromosomes, background genetic variability, and family heterogeneity. Lorenz curves and Gini coefficients are suggested for assessing the inequality of the contribution of different marked effects to genetic variability. Classical and Bayesian methods are presented. The Bayesian approach includes a Markov chain Monte Carlo implementation. The generality and flexibility of the Bayesian method is illustrated when a Lorenz curve is to be inferred. PMID- 12586722 TI - Bayesian analysis of genetic differentiation between populations. AB - We introduce a Bayesian method for estimating hidden population substructure using multilocus molecular markers and geographical information provided by the sampling design. The joint posterior distribution of the substructure and allele frequencies of the respective populations is available in an analytical form when the number of populations is small, whereas an approximation based on a Markov chain Monte Carlo simulation approach can be obtained for a moderate or large number of populations. Using the joint posterior distribution, posteriors can also be derived for any evolutionary population parameters, such as the traditional fixation indices. A major advantage compared to most earlier methods is that the number of populations is treated here as an unknown parameter. What is traditionally considered as two genetically distinct populations, either recently founded or connected by considerable gene flow, is here considered as one panmictic population with a certain probability based on marker data and prior information. Analyses of previously published data on the Moroccan argan tree (Argania spinosa) and of simulated data sets suggest that our method is capable of estimating a population substructure, while not artificially enforcing a substructure when it does not exist. The software (BAPS) used for the computations is freely available from http://www.rni.helsinki.fi/~mjs. PMID- 12586723 TI - Bounds on the minimum number of recombination events in a sample history. AB - Recombination is an important evolutionary factor in many organisms, including humans, and understanding its effects is an important task facing geneticists. Detecting past recombination events is thus important; this article introduces statistics that give a lower bound on the number of recombination events in the history of a sample, on the basis of the patterns of variation in the sample DNA. Such lower bounds are appropriate, since many recombination events in the history are typically undetectable, so the true number of historical recombinations is unobtainable. The statistics can be calculated quickly by computer and improve upon the earlier bound of Hudson and Kaplan 1985. A method is developed to combine bounds on local regions in the data to produce more powerful improved bounds. The method is flexible to different models of recombination occurrence. The approach gives recombination event bounds between all pairs of sites, to help identify regions with more detectable recombinations, and these bounds can be viewed graphically. Under coalescent simulations, there is a substantial improvement over the earlier method (of up to a factor of 2) in the expected number of recombination events detected by one of the new minima, across a wide range of parameter values. The method is applied to data from a region within the lipoprotein lipase gene and the amount of detected recombination is substantially increased. Further, there is strong clustering of detected recombination events in an area near the center of the region. A program implementing these statistics, which was used for this article, is available from http://www.stats.ox.ac.uk/mathgen/programs.html. PMID- 12586724 TI - Estimating ancestral population sizes and divergence times. AB - This article presents a new method for jointly estimating species divergence times and ancestral population sizes. The method improves on previous ones by explicitly incorporating intragenic recombination, by utilizing orthologous sequence data from closely related species, and by using a maximum-likelihood framework. The latter allows for efficient use of the available information and provides a way of assessing how much confidence we should place in the estimates. I apply the method to recently collected intergenic sequence data from humans and the great apes. The results suggest that the human-chimpanzee ancestral population size was four to seven times larger than the current human effective population size and that the current human effective population size is slightly >10,000. These estimates are similar to previous ones, and they appear relatively insensitive to assumptions about the recombination rates or mutation rates across loci. PMID- 12586725 TI - Multitrait fine mapping of quantitative trait loci using combined linkage disequilibria and linkage analysis. AB - A novel multitrait fine-mapping method is presented. The method is implemented by a model that treats QTL effects as random variables. The covariance matrix of allelic effects is proportional to the IBD matrix, where each element is the probability that a pair of alleles is identical by descent, given marker information and QTL position. These probabilities are calculated on the basis of similarities of marker haplotypes of individuals of the first generation of genotyped individuals, using "gene dropping" (linkage disequilibrium) and transmission of markers from genotyped parents to genotyped offspring (linkage). A small simulation study based on a granddaughter design was carried out to illustrate that the method provides accurate estimates of QTL position. Results from the simulation also indicate that it is possible to distinguish between a model postulating one pleiotropic QTL affecting two traits vs. one postulating two closely linked loci, each affecting one of the traits. PMID- 12586726 TI - Polymorphism and divergence for island-model species. AB - Estimates of the scaled selection coefficient, gamma of Sawyer and Hartl, are shown to be remarkably robust to population subdivision. Estimates of mutation parameters and divergence times, in contrast, are very sensitive to subdivision. These results follow from an analysis of natural selection and genetic drift in the island model of subdivision in the limit of a very large number of subpopulations, or demes. In particular, a diffusion process is shown to hold for the average allele frequency among demes in which the level of subdivision sets the timescale of drift and selection and determines the dynamic equilibrium of allele frequencies among demes. This provides a framework for inference about mutation, selection, divergence, and migration when data are available from a number of unlinked nucleotide sites. The effects of subdivision on parameter estimates depend on the distribution of samples among demes. If samples are taken singly from different demes, the only effect of subdivision is in the rescaling of mutation and divergence-time parameters. If multiple samples are taken from one or more demes, high levels of within-deme relatedness lead to low levels of intraspecies polymorphism and increase the number of fixed differences between samples from two species. If subdivision is ignored, mutation parameters are underestimated and the species divergence time is overestimated, sometimes quite drastically. Estimates of the strength of selection are much less strongly affected and always in a conservative direction. PMID- 12586727 TI - A diffusion approximation for selection and drift in a subdivided population. AB - The population-genetic consequences of population structure are of great interest and have been studied extensively. An area of particular interest is the interaction among population structure, natural selection, and genetic drift. At first glance, different results in this area give very different impressions of the effect of population subdivision on effective population size (N(e)), suggesting that no single value of N(e) can completely characterize a structured population. Results presented here show that a population conforming to Wright's island model of subdivision with genic selection can be related to an idealized panmictic population (a Wright-Fisher population). This equivalent panmictic population has a larger size than the actual population; i.e., N(e) is larger than the actual population size, as expected from many results for this type of population structure. The selection coefficient in the equivalent panmictic population, referred to here as the effective selection coefficient (s(e)), is smaller than the actual selection coefficient (s). This explains how the fixation probability of a selected allele can be unaffected by population subdivision despite the fact that subdivision increases N(e), for the product N(e)s(e) is not altered by subdivision. PMID- 12586728 TI - Estimating effective population size and migration rates from genetic samples over space and time. AB - In the past, moment and likelihood methods have been developed to estimate the effective population size (N(e)) on the basis of the observed changes of marker allele frequencies over time, and these have been applied to a large variety of species and populations. Such methods invariably make the critical assumption of a single isolated population receiving no immigrants over the study interval. For most populations in the real world, however, migration is not negligible and can substantially bias estimates of N(e) if it is not accounted for. Here we extend previous moment and maximum-likelihood methods to allow the joint estimation of N(e) and migration rate (m) using genetic samples over space and time. It is shown that, compared to genetic drift acting alone, migration results in changes in allele frequency that are greater in the short term and smaller in the long term, leading to under- and overestimation of N(e), respectively, if it is ignored. Extensive simulations are run to evaluate the newly developed moment and likelihood methods, which yield generally satisfactory estimates of both N(e) and m for populations with widely different effective sizes and migration rates and patterns, given a reasonably large sample size and number of markers. PMID- 12586729 TI - Identification of a functional domain within the essential core of histone H3 that is required for telomeric and HM silencing in Saccharomyces cerevisiae. AB - Fourteen novel single-amino-acid substitution mutations in histone H3 that disrupt telomeric silencing in Saccharomyces cerevisiae were identified, 10 of which are clustered within the alpha1 helix and L1 loop of the essential histone fold. Several of these mutations cause derepression of silent mating locus HML, and an additional subset cause partial loss of basal repression at the GAL1 promoter. Our results identify a new domain within the essential core of histone H3 that is required for heterochromatin-mediated silencing. PMID- 12586730 TI - Mosaicism of Solid Gold supports the causality of a noncoding A-to-G transition in the determinism of the callipyge phenotype. AB - To identify the callipyge mutation, we have resequenced 184 kb spanning the DLK1 , GTL2-, PEG11-, and MEG8-imprinted domain and have identified an A-to-G transition in a highly conserved dodecamer motif between DLK1 and GTL2. This was the only difference found between the callipyge (CLPG) allele and a phylogenetically closely related wild-type allele. We report that this SNP is in perfect association with the callipyge genotype. The demonstration that Solid Gold-the alleged founder ram of the callipyge flock-is mosaic for this SNP virtually proves the causality of this SNP in the determinism of the callipyge phenotype. PMID- 12586731 TI - Negative role of cAMP-dependent protein kinase A in RANTES-mediated transcription of proinflammatory mediators through Raf. AB - The chemokine RANTES (regulated on activation normal T cell expressed and secreted) is expressed in several inflammatory diseases of the central nervous system and is a powerful stimulus for astrocyte production of proinflammatory mediators. The mechanism of RANTES-mediated astrocyte activation was investigated. RANTES stimulation decreased both intracellular cyclic AMP (cAMP) levels and cAMP-dependent protein kinase A (PKA) activity in cultures of primary mouse astrocytes. H-89, a potent inhibitor of PKA, mimicked RANTES-mediated chemokine and cytokine transcription. RANTES treatments activated Raf-1 kinase activity, and conversely a dominant negative Raf and a Raf-1 inhibitor blocked RANTES-induced chemokine transcription. Transfection with a constitutively active Raf was sufficient to induce transcription of proinflammatory mediators. The combined data indicate that Raf-1 is required for RANTES-mediated astrocyte activation. Decreases of cAMP and PKA activity contributed to the transcription of proinflammatory mediators by cross-talk with the Raf-1/mitogen-activated protein kinase pathway. The results identify an upstream signaling pathway for amplification of proinflammatory mediators in the central nervous system. PMID- 12586732 TI - Hyperosmolarity and CD95L trigger CD95/EGF receptor association and tyrosine phosphorylation of CD95 as prerequisites for CD95 membrane trafficking and DISC formation. AB - The mechanisms underlying CD95 ligand (CD95L)- and hyperosmolarity-induced activation of the CD95 system [Reinehr, R., Graf, D., Fischer, R., Schliess, F., and Haussinger, D. (2002) Hepatology 36, 602-614] as initial steps of apoptosis were studied. Hyperosmotic exposure (405 mosmol/l) of rat hepatocytes induced within 1 min oxidative stress and antioxidant-sensitive activation of the epidermal growth factor receptor (EGFR) and c-Jun-N-terminal-kinase (JNK). After 30 min of hyperosmotic exposure EGFR associated with CD95 and CD95 became tyrosine phosphorylated. Inhibition of JNK or protein kinase C (PKC) had no effect on EGFR phosphorylation but abolished CD95/EGFR association, CD95-tyrosine phosphorylation, membrane targeting, and Fas-associated death domain/caspase 8 recruitment to CD95 [death-inducing signaling complex (DISC) formation]. Inhibition of EGFR tyrosine kinase activity prevented CD95 tyrosine phosphorylation and DISC formation but not hyperosmolarity-induced EGFR phosphorylation and EGFR association with CD95. Tyrosine-phosphorylated CD95 was enriched in the plasma membrane. All maneuvers preventing CD95 tyrosine phosphorylation inhibited CD95 membrane trafficking and DISC formation. Stimulation of EGFR by EGF induced EGFR phosphorylation but no association with CD95 or CD95 phosphorylation. Addition of CD95L also induced EGFR and JNK activation, EGFR/CD95 association, CD95 tyrosine phosphorylation, DISC formation, and CD95 membrane targeting with an inhibitor sensitivity profile similar to that of hyperosmotic CD95 activation, except that inhibition of PKC was ineffective. The data suggest that moderate hyperosmolarity or CD95L trigger oxidative stress and EGFR activation followed by a JNK-dependent EGFR/CD95association and CD95 tyrosine phosphorylation, probably through EGFR tyrosine kinase activity. This provides a signal for CD95 membrane trafficking and DISC formation. PMID- 12586733 TI - A therapeutic role for cyclooxygenase-2 inhibitors in a transgenic mouse model of amyotrophic lateral sclerosis. AB - Recent studies indicate that the proinflammatory enzyme cyclooxygenase (COX)-2, an enzyme involved in inflammatory cascades but also normal neuronal activities, is elevated in the brain and spinal cord of amyotrophic lateral sclerosis (ALS) patients and ALS mouse model systems. On the basis of this evidence, we explored the impact of COX-2 inhibition on the onset and progression of ALS-like disease in the G93A human superoxide dismutase (SOD)1 mouse model of ALS. We found that prophylactic administration of nimesulide, a preferential COX-2 inhibitor, in the feed resulted in a significant delay in the onset of ALS type motor impairment. This delay of ALS symptomatology temporally overlapped with the inhibition of prostaglandin E2 elevation in the spinal cord of SOD1-G93A transgenic mice relative to untreated SOD1-G93A controls. This study strongly supports a role for COX-2 in the pathophysiology of ALS and provides the first experimental evidence that prophylactic treatment with COX-2 inhibitors can significantly delay the onset of motor dysfunction in the SOD1-G93A transgenic mouse model of ALS. PMID- 12586734 TI - Anoxia and reoxygenation of human endothelial cells decrease ceramide glucosyltransferase expression and activates caspases. AB - Endothelial oxidative stress induces cellular activation and sometimes death. Endothelial death can occur via necrosis or apoptosis. Understanding the mechanisms involved in cellular activation and death may lead to therapeutics designed to increase death or preserve cellular function. In the present study, brief periods of anoxia (3 h) followed by varying lengths of reoxygenation (0-5 h) lead to a time-dependent increase in human umbilical vein endothelial cell (HUVEC) caspase activity. Furthermore, ROCK-1 cleavage, which is dependent on caspase-3 activity, was also increased in cells undergoing oxidative stress compared with normoxic cells. Microarray data demonstrated that glucosylceramide synthase (GCS; glucosylceramide transferase), but not acid sphingomyelinase, was modulated by anoxia and reoxygenation. We confirmed that GCS mRNA and protein expression were significantly decreased in a time-dependent fashion following oxidative stress by real-time polymerase chain reaction and Western blot, respectively. Treatment of normoxic cells with the GCS-specific inhibitor, D,L threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), increased caspase activity to the same degree as cells undergoing oxidative stress. Fumonisin B1, the N-acyl-sphinganine dehydrogenase (e.g., ceramide synthase) inhibitor significantly attenuated caspase activity in HUVECs undergoing oxidative stress. These data suggest that alterations in GCS expression following brief periods of oxidative stress in human endothelial cells lead to increased caspase activity. PMID- 12586735 TI - The phosphatidylinositol 3-kinase inhibitor LY294002 potently blocks K(V) currents via a direct mechanism. AB - Voltage-dependent K+ (Kv) channels negatively regulate Ca2+ entry into pancreatic beta-cells by repolarizing glucose-stimulated action potentials. A role for phosphatidylinositol 3-kinase (PI3K) modulation of Kv channel function was investigated using the PI3K inhibitors wortmannin and LY294002, and LY303511, a negative control compound with respect to PI3K activity. In MIN6 insulinoma cells, wortmannin (100 nM) had no effect on whole-cell outward K+ currents, but LY294002 and LY303511 reversibly blocked currents in a dose-dependent manner (IC50=9.0+/-0.7 microM and 64.6+/-9.1 microM, respectively). Western blotting confirmed the specific inhibitory effects of LY294002 and wortmannin on insulin stimulated PI3K activity. Kv currents in rat beta-cells at near physiological temperatures were inhibited 92% by 25 microM LY294002. Kv2.1 and Kv1.4 are highly expressed in beta-cells, and in Kv2.1-transfected tsA201 cells, 50 microM LY294002 and 100 microM LY303511 reversibly inhibited currents by 99% and 41%, respectively. In Kv1.4-transfected tsA201 cells, 50 microM LY294002 reduced the inactivation time constant from 73 to 18 ms. The insulinotropic properties of LY294002 and its effects in other excitable cells may be caused by inhibition of Kv currents rather than PI3K antagonism. Furthermore, LY294002 may represent a novel structure from which future Kv channel blockers may be developed. PMID- 12586736 TI - Enhanced angiogenesis and improvement of neuropathy by cotransfection of human hepatocyte growth factor and prostacyclin synthase gene. AB - The current therapeutic angiogenesis strategy to treat ischemic disease by using angiogenic growth factors has been limited to use of a single gene. However, as vasodilator substances such as prostacyclin are widely used for the treatment of peripheral arterial disease, it might be useful to combine angiogenesis with vasodilation of new vessels. In a mouse hind limb ischemia model, cotransfection of the hepatocyte growth factor (HGF) gene with the prostacyclin synthase gene demonstrated a further increase in blood flow and capillary density compared with a single gene. Even in the rabbit ischemia model, cotransfection of HGF plasmid with the prostacyclin synthase gene demonstrated a further increase in angiogenic activity compared with HGF alone. Because peripheral neuropathy due to diabetes is common for significant morbidity, we examined the hypothesis that experimental diabetic neuropathy can be reversed by HGF and prostacyclin synthase genes. Severe peripheral neuropathy, characterized by significant slowing of nerve conduction velocity compared with nondiabetic control animals, was ameliorated. Overall, cotransfection of the prostacyclin synthase and HGF genes is more effective than single-gene transfection to stimulate angiogenesis, and it significantly improved neuropathy. These data provide important information relating to the clinical application of therapeutic angiogenesis to treat peripheral arterial disease. PMID- 12586737 TI - Decreased complex III activity in mitochondria isolated from rat heart subjected to ischemia and reperfusion: role of reactive oxygen species and cardiolipin. AB - Reactive oxygen species (ROS) are considered an important factor in ischemia reperfusion injury to cardiac myocites. We have examined the effects of ischemia (30 min) and ischemia followed by reperfusion (15 min) of rat hearts on the activity of complex III and on the cardiolipin content in isolated mitochondria. Mitochondrial production of H2O2 and lipid peroxidation was also measured. The capacity of mitochondria to produce both H2O2 and lipid peroxidation increased upon reperfusion. The activity of complex III was 22% and 46% lower in ischemic and reperfused rat heart mitochondria, respectively, than that of controls. These changes in complex III activity were associated to parallel changes in state 3 respiration. The mitochondrial content of cardiolipin, which is required for optimal activity of complex III, decreased by 28% and by 50% as a function of ischemia and reperfusion, respectively. The lower complex III activity in mitochondria from reperfused rat hearts could be completely restored to the level of normal hearts by exogenously added cardiolipin. It is proposed that the loss of complex III activity in reperfused rat hearts can be mainly ascribed to a loss of cardiolipin content, due to oxidative attack by oxygen free radicals. PMID- 12586738 TI - The p54 cleaved form of the tyrosine kinase Lyn generated by caspases during BCR induced cell death in B lymphoma acts as a negative regulator of apoptosis. AB - Engagement of the B cell receptor antigen (BCR) triggers apoptosis on immature B cell lines. We report here that BCR triggering leads to caspase activation followed by Lyn cleavage and induction of apoptosis. The cleavage process is mitochondrion-dependent and involves caspases 9 and 7. Stable expression of the cleaved form of Lyn (Lyn-Delta-N) in Ramos B cells impairs BCR-mediated apoptosis as judged by loss of Delta(psi)m, caspase activation and PARP cleavage. Activation of the main survival pathways upon BCR-triggering was unaltered in both cell variants. However, the PI3-K inhibitor Ly294002 resensitizes Lyn-Delta N cells to apoptosis. Selected cDNA expression arrays revealed that anti-IgM modulates the expression of approximately 20 genes in both cell variants. Among them, only c-Myc was found to be differentially regulated, which suggests a role for c-Myc in the B cell apoptotic response. Interestingly, c-Myc expression decreased more rapidly in Lyn-Delta-N compared with Lyn-WT cells during the first hours of anti-IgM stimulation. Nevertheless, rapid down-regulation of c-Myc following BCR engagement seems to correlate with the resistance of B cells to apoptosis. Thus, the soluble form of Lyn generated by caspases following BCR triggering acts as an inhibitor of B lymphocyte death likely through the modulation of c-Myc expression. PMID- 12586739 TI - From energy store to energy flux: a study in creatine kinase-deficient fast skeletal muscle. AB - Fast-twitch skeletal muscle of mice deficient in cytosolic and mitochondrial creatine kinase isoforms (CK-/-) lack burst activity but can sustain prolonged contractile activity, suggesting that adaptive mechanisms can regulate local adenine nucleotide turnover. We investigated whether direct energy and signal channeling between mitochondria and sarcoplasmic reticulum (SR) or myofilaments may exist that compensate for the lack of CK isoenzymes. Oxidative capacity of fast-twitch muscle was increased twofold in CK-/- mice. Energy cross talk between organelles was studied in muscle fibers with permeabilized sarcolemma. Energy supply to SR was estimated by analyzing the tension transient induced by caffeine and energy supply to myofilaments was estimated by the relaxation of rigor tension, both under different conditions of energy supply. In normal mice, ATP directly produced by mitochondria was not able to sustain calcium uptake and to relax rigor tension as efficiently as ATP produced by bound CK. However, in CK-/- mice, mitochondria ability to provide ATP for calcium uptake and relaxation of rigor tension was dramatically enhanced, suggesting a direct ATP/ADP channeling between sites of energy production mitochondria) and energy utilization in CK-/- mice. These results demonstrate two possible patterns of energy transport in muscle cells: energy store with phosphocreatine and energy flux through mitochondria. PMID- 12586740 TI - VEGF165 transfection decreases postischemic NF-kappa B-dependent myocardial reperfusion injury in vivo: role of eNOS phosphorylation. AB - Endothelial nitric oxide synthase (eNOS) phosphorylation increases nitric oxide formation, for example, after VEGF stimulation. We investigated whether nitric oxide formed after overexpression of VEGF or of phosphomimetic eNOS (S1177D) affects PMN-induced myocardial detriment after ischemia and reperfusion. Pigs (n=8 per group) were subjected to percutaneous liposome-based gene transfer by retroinfusion of the anterior interventricular vein 48 h before LAD occlusion (60 min) and reperfusion (24 h). Thereafter, regional myocardial function was assessed as subendocardial segment shortening (SES), and infarct size was determined. Tissue from the infarct region, the noninfarcted area at risk, and a control region was analyzed for NF-kappaB activation (EMSA), tumor necrosis factor (TNF)-alpha, and E-selectin mRNA and infiltration of polymorphonuclear neutrophils (PMN). L-NAME was applied in one group of VEGF-transfected animals. NF-kappaB activition, PMN infiltration in the infarct region, and AAR were reduced after transfection of VEGF or eNOS S1177D, but not after VEGF+L-NAME coapplication. Infarct size decreased, whereas SES improved after either VEGF or eNOS S1177D transfection, an effect inhibited by L-NAME coapplication. Retroinfusion of liposomal VEGF cDNA reduces NF-kappaB-dependent postischemic inflammation and subsequent myocardial reperfusion injury, an effect mediated at least in part by enhanced eNOS phosphorylation. PMID- 12586741 TI - Protection against lipopolysaccharide-induced endothelial dysfunction in resistance and conduit vasculature of iNOS knockout mice. AB - Endothelial dysfunction is a characteristic of, and may be pathogenic in, inflammatory cardiovascular diseases, including sepsis. The mechanism underlying inflammation-induced endothelial dysfunction may be related to the expression and activity of inducible nitric oxide synthase (iNOS). This possibility was investigated in isolated resistance (mesenteric) and conduit (aorta) arteries taken from lipopolysaccharide (LPS)-treated (12.5 mg/kg i.v.) or saline-treated iNOS knockout (KO) and wild-type (WT) mice. LPS pretreatment (for 15 h, but not 4 h) profoundly suppressed responses to acetylcholine (ACh) and significantly reduced sensitivity to the NO donor spermine-NONOate (SPER-NO) in aorta and mesenteric arteries of WT mice. This effect was temporally associated with iNOS protein expression in both conduit and resistance arteries and with a 10-fold increase in plasma NOx levels. In contrast, no elevation of plasma NOx was observed in LPS-treated iNOS KO animals, and arteries dissected from these animals did not express iNOS or display hyporeactivity to ACh or SPER-NO. The mechanism underlying this phenomenon may be suppression of eNOS expression, as observed in arteries of WT animals, that was absent in arteries of iNOS KO animals. These results clearly demonstrate that iNOS induction plays an integral role in mediation of the endothelial dysfunction associated with sepsis in both resistance and conduit arteries. PMID- 12586742 TI - Green tea polyphenol epigallocatechin-3 gallate induces apoptosis of proliferating vascular smooth muscle cells via activation of p53. AB - Green tea polyphenols (GTPs), which possess antioxidant properties, have been shown to inhibit the development of atherosclerotic lesions. Epigallocatechin-3 gallate (EGCG), the most abundant GTP, displays antiproliferative effects in a variety of cell types. Here, we examined the effects of GTPs on aortic smooth muscle cell (SMC) proliferation. Treatment with a GTP mixture or EGCG at a dose of 40 to 50 microg/ml slowed SMC growth, while at a higher dose of 80 microg/ml EGCG also induced cell death as judged by TUNEL assay. Apoptosis was mainly observed in proliferating SMCs in subconfluent cultures; whereas at higher confluency, cell viability was largely unaffected. Treatment with 80 microg/ml EGCG induced the tumor suppressor p53, which was functional as judged by activation of the target cyclin-dependent kinase inhibitor p21CIP1. Inhibition of p53 activity with a dominant negative mutant reduced cell death. The increase in p53 protein was due to increased stability. EGCG also induced functional nuclear factor-kappaB (NF-kappaB) complexes, and inhibition of this activity reduced the extent of cell death. Thus, EGCG inhibits growth and induces death of SMCs in a p53- and NF-kappaB-dependent manner. These results provide evidence for a new molecular mechanism whereby green tea polyphenols inhibit SMC proliferation and function to prevent the development of atherosclerosis. PMID- 12586743 TI - Attenuated growth of breast-fed children exposed to increased concentrations of methylmercury and polychlorinated biphenyls. AB - Breast-feeding has been linked to slowed postnatal growth. Although the basis for this "weanling's dilemma" is unclear, environmental contaminants in human milk may be of relevance. We studied a Faroese birth cohort of 182 singleton children, born at term in 1994-95. Concentrations of mercury in cord blood and of polychlorinated biphenyls in maternal milk were measured, and duration of breast feeding was recorded. At 18 months, children who had been exclusively breast-fed for at least 6 months weighed 0.59 kg less [95% confidence interval (CI) = 0.03, 1.16 kg] and were 1.50 cm [95% CI = 0.52, 2.47 cm] shorter than those not breast fed. However, calculated transfer of contaminants from human milk fully explained the attenuated growth. Irrespective of duration of breast-feeding, a doubling of the mercury concentration in cord blood was associated with a decrease in weight at 18 months by 0.19 kg (95% CI = 0.03, 0.35 kg) and in height by 0.26 cm (95% CI = -0.02, 0.55 cm). Weight and height at 42 months showed the same tendencies, but the main effect occurred before 18 months of age. Thus, in communities with increased contaminant exposures, risks associated with lactational transfer of toxicants to the infant must be considered when judging the benefits of prolonged breast-feeding. PMID- 12586744 TI - Cytochrome P450 2C9-derived epoxyeicosatrienoic acids induce angiogenesis via cross-talk with the epidermal growth factor receptor (EGFR). AB - Cytochrome P450 (CYP) epoxygenase products, such as 11,12-epoxyeicosatrienoic acid (EET), stimulate endothelial cell proliferation. We set out to identify the signal transduction cascade linking EET generation to enhanced proliferation and angiogenesis. In human endothelial cells overexpressing CYP 2C9, cell number was increased compared with control cells and was inhibited by the CYP 2C9 inhibitor, sulfaphenazole. CYP 2C9 overexpression was associated with the activation of Akt and an increase in cyclin D1 expression, effects that were abolished by the epidermal growth factor (EGF) receptor inhibitor, AG1478, which also prevented the CYP 2C9-induced increase in cell proliferation. Stimulation of EGF receptor overexpressing cells with 11,12-EET or transfection of these cells with CYP 2C9 enhanced the tyrosine phosphorylation of the EGF receptor. Endothelial tube formation in a fibrin gel was significantly enhanced (6-fold) in CYP 2C9 overexpressing cells and was comparable with the tube formation induced by EGF. In the chick chorioallantoic membrane, 11,12-EET stimulated vessel formation (3.5 fold) and induced vessel convergence, an effect that was abolished by cotreatment with either an EGF receptor-neutralizing antibody or AG1478. These results indicate that CYP 2C9-derived EETs stimulate angiogenesis by a mechanism involving the activation of the EGF receptor. PMID- 12586745 TI - Immortalization of bovine capillary endothelial cells by hTERT alone involves inactivation of endogenous p16INK4A/pRb. AB - Recent studies show that stable expression of the human telomerase catalytic subunit, hTERT, alone can lead several types of normal human somatic cells to bypass replicative senescence and become immortal. The molecular mechanisms by which telomerase immortalizes cells are not fully understood, although a key function of telomerase is to maintain a critical length of telomeres in order to preserve the stability and integrity of the genome. Here we report that stable transfection of hTERT alone was sufficient to allow bovine capillary endothelial (BCE) cells to bypass senescence and acquire immortality. Surprisingly, telomere lengths in these stable transfectants were progressively shortened during an increasing number of population doublings (PDLs), despite high telomerase activity. The expression of the cyclin-dependent kinase inhibitors (CDKIs) p16INK4A and p21CIP1/WAF1 was concomitantly repressed, and the retinoblastoma protein (pRb) was maintained in a hyperphosphorylated state in the telomerase expressing cells. Re-expression of p16INK4A in these cells by either treatment with a demethylating agent or by adenovirus-mediated expression reinduced a senescence-like phenotype, suggesting that the inactivation of p16INK4A was due to DNA methylation and was crucial for the immortalization process. In agreement with this finding, the expression levels of the prototypic DNA methyltransferase DNMT1 were elevated in the hTERT-positive cells. PMID- 12586746 TI - Genotype and age influence the effect of caloric intake on mortality in mice. AB - Long-term caloric restriction (CR) has been repeatedly shown to increase life span and delay the onset of age-associated pathologies in laboratory mice and rats. The purpose of the current study was to determine whether the CR-associated increase in life span occurs in all strains of mice or only in some genotypes and whether the effects of CR and ad libitum (AL) feeding on mortality accrue gradually or are rapidly inducible and reversible. In one experiment, groups of male C57BL/6, DBA/2, and B6D2F1 mice were fed AL or CR (60% of AL) diets beginning at 4 months of age until death. In the companion study, separate groups of mice were maintained chronically on AL or CR regimens until 7, 17, or 22-24 months of age, after which, half of each AL and CR group was switched to the opposite regimen for 11 wk. This procedure yielded four experimental groups for each genotype, namely AL-->AL, AL-->CR, CR-->CR, and CR-->AL, designated according to long-term and short-term caloric regimen, respectively. Long-term CR resulted in increased median and maximum life span in C57BL/6 and B6D2F1 mice but failed to affect either parameter in the DBA/2 mice. The shift from AL-->CR increased mortality in 17- and 24-month-old mice, whereas the shift from CR-->AL did not significantly affect mortality of any age group. Such increased risk of mortality following implementation of CR at older ages was evident in all three strains but was most dramatic in DBA/2 mice. Results of this study indicate that CR does not have beneficial effects in all strains of mice, and it increases rather than decreases mortality if initiated in advanced age. PMID- 12586747 TI - Complementary roles of glycoprotein VI and alpha2beta1 integrin in collagen induced thrombus formation in flowing whole blood ex vivo. AB - Platelets interact vigorously with subendothelial collagens that are exposed by injury or pathological damage of a vessel wall. The collagen-bound platelets trap other platelets to form aggregates, and they expose phosphatidylserine (PS) required for coagulation. Both processes are implicated in the formation of vaso occlusive thrombi. We previously demonstrated that the immunoglobulin receptor glycoprotein VI (GPVI), but not integrin alpha2beta1, is essential in priming platelet-collagen interaction and subsequent aggregation. Here, we report that these receptors have yet a complementary function in ex vivo thrombus formation during perfusion of whole blood over collagen. With mice deficient in GPVI or blocking antibodies, we found that GPVI was indispensable for collagen-dependent Ca2+ mobilization, exposure of PS, and aggregation of platelets. Deficiency of integrin beta1 reduces the GPVI-evoked responses but still allows the formation of loose platelet aggregates. By using mice deficient in G(alpha)q or specific thromboxane A2 and ADP antagonists, we show that these autocrine agents mediated aggregation but not collagen-induced Ca2+ mobilization or PS exposure. Collectively, these data indicate that integrin alpha2beta1 facilitates the central function of GPVI in the platelet activation processes that lead to thrombus formation, whereas the autocrine thromboxane A2 and ADP serve mainly to trigger aggregate formation. PMID- 12586748 TI - HDL counterbalance the proinflammatory effect of oxidized LDL by inhibiting intracellular reactive oxygen species rise, proteasome activation, and subsequent NF-kappaB activation in smooth muscle cells. AB - Oxidized low-density lipoproteins (oxLDL) exhibit proinflammatory properties and play a role in atherosclerosis plaque formation, rupture, and subsequent thrombosis. OxLDL alter the activity of the transcription factor NF-kappaB that is involved in the expression of immune and inflammatory genes. In contrast, high density lipoproteins (HDL) are anti-atherogenic and exhibit anti-inflammatory properties. This work aimed to investigate how oxLDL activate NF-kappaB and whether and how HDL may prevent NF-kappaB activation. In cultured rabbit smooth muscle cells, mitogenic concentrations of mildly oxLDL trigger a rapid and transient NF-kappaB activation, which is strongly inhibited by HDL. Growth factors, phosphatidylinositol 3-kinase/Akt, and sphingosine kinase pathways are not implicated in the oxLDL-induced NF-kappaB activation and are not targets of HDL. OxLDL induce reactive oxygen species (ROS) generation and proteasome activation, which are implicated in NF-kappaB activation, as suggested by the inhibitory effect of the antioxidants N-acetyl-L-cysteine and pyrrolidinedithiocarbamate and the proteasome inhibitor PSI. HDL were able to prevent the intracellular ROS rise triggered by oxLDL or H2O2, thereby inhibiting the subsequent proteasome activation, IkappaB degradation, and NF-kappaB activation. In conclusion, the oxLDL-induced NF-kappaB activation involves ROS generation and proteasome activation, both events being inhibited by HDL. This 'antioxidant' and potentially anti-inflammatory effect of HDL may participate in their general anti-atherogenic properties. PMID- 12586749 TI - Inhibition of F-Box protein p45(SKP2) expression and stabilization of cyclin dependent kinase inhibitor p27(KIP1) in vitamin D analog-treated cancer cells. AB - Treatment of cancer cells with 1,25-dihydroxyvitamin D3 [1,25(OH)(2)D(3)] or its analogs induces growth arrest and expression of the cyclin-dependent kinase inhibitor p27(KIP1). Although 1,25(OH)(2)D(3) transiently enhances p27(kip1) gene transcription in some cells, its effects on p27(KIP1) protein levels are generally more gradual and sustained. This suggests that 1,25(OH)(2)D(3) treatment may be stabilizing p27(KIP1) protein, which is sensitive to modification by the SCF(SKP2) protein ubiquitin ligase and proteosomal degradation. Here, we show that treatment of AT-84 head and neck squamous carcinoma cells with the 1,25(OH)(2)D(3) analog EB1089 increases p27(KIP1) protein levels without significantly affecting expression of its mRNA. EB1089 treatment repressed expression of mRNAs encoding the F-box protein p45(SKP2), a marker of poor head and neck cancer prognosis, and the cyclin kinase subunit CKS1, which is essential for targeting p45(SKP2) to p27(KIP1). This coincided with a reduction of total p45(SKP2) protein, and p45(SKP2) associated with p27(KIP1). Consistent with these findings, turnover of p27(KIP1) protein was strongly inhibited in the presence of EB1089. A similar reduction in p45(SKP2) expression and stabilization of p27(KIP1) protein was observed in 1,25(OH)(2)D(3) sensitive UF-1 promyelocytic leukemia cells, which also respond by transiently increasing p27(kip1) gene transcription. Our results reveal that 1,25(OH)(2)D(3) analogs increase levels of p27(KIP1) in different cell types by inhibiting expression of SCF(SKP2) subunits and reducing turnover of p27(KIP1) protein. PMID- 12586750 TI - The role of ghrelin and growth hormone secretagogues receptor on rat adipogenesis. AB - Recent research progress indicates a close link between ghrelin, a natural ligand of GH secretagogues receptor (GHS-R), and both the metabolic balance and body composition. To clarify the involvement of ghrelin and GHS-R in the process of adipogenesis, we measured the expression of GHS-R and peroxisome proliferator activated receptor gamma 2 (PPAR-gamma 2) mRNA in rat adipocytes using semiquantitative RT-PCR methods. The levels of GHS-R mRNA increased by up to 4 fold in adipose tissue from epididymal and parametrial regions as the rat aged from 4-20 wk and were significantly elevated during the differentiation of preadipocytes in vitro. Ghrelin (10(-8) M for 10 d) stimulated the activity of glycerol-3-phosphate dehydrogenase and the differentiation of rat preadipocytes in vitro. Ghrelin treatment also significantly increased the levels of PPAR-gamma 2 mRNA in primary cultured rat differentiated adipocytes. In addition, isoproterenol (10(-8) M, 40 min)-stimulated lipolysis was significantly reduced by simultaneous ghrelin treatment in a dose-dependent manner in vitro. In conclusion, the expression of GHS-R in rat adipocytes increases with the age and during adipogenesis. Ghrelin in vitro stimulates the differentiation of preadipocytes and antagonizes lipolysis. Ghrelin may therefore play an important role in the process of adipogenesis in rats. PMID- 12586751 TI - Proopiomelanocortin-derived peptides in rat cerebrospinal fluid and hypothalamic extracts: evidence that secretion is regulated with respect to energy balance. AB - Regulation of proopiomelanocortin (POMC) is an important means of controlling the central melanocortin system. It has never been established whether the spectrum of POMC-derived peptides synthesized and secreted from the hypothalamus is altered in response to changes in energy homeostasis in vivo. To monitor secretion, we analyzed peptide content of rat cerebrospinal fluid. Strikingly, both the POMC precursor and ACTH were readily detected. Moreover, levels of both were lower in samples from obese Zucker rats (fa/fa) vs. lean Zucker rats (+/+, fa/+) and from fasted vs. fed rats, whereas alpha MSH could not be detected. POMC levels were also decreased in hypothalamic extracts from obese and fasted animals. In contrast, despite being the most predominant peptide in extracts, alpha MSH levels were not significantly changed in any of the rat models. The ratio of precursor to derived peptides in cerebrospinal fluid was significantly higher in obese vs. lean and fed vs. fasted rats, indicating that secretion of POMC-derived peptides is differentially down-regulated during negative energy balance. In contrast to peptide analysis, we found that POMC gene expression was not significantly decreased in fasted rat hypothalami. We conclude that regulation of peptide secretion is an important mechanism by which the POMC system is controlled. PMID- 12586752 TI - Expression of aryl hydrocarbon receptor and aryl hydrocarbon receptor nuclear translocator messenger ribonucleic acids and proteins in rat and human testis. AB - Dioxins, e.g. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), use the aryl hydrocarbon receptor (AHR)/aryl hydrocarbon receptor nuclear translocator (ARNT) receptor complex to mediate their toxic actions. In addition to interaction with environmental pollutants, several transcription factors, steroid receptors, and growth factors are capable interacting with the AHR/ARNT complex, which suggests a constitutive role for the receptor complex. The testis has been reported to be among the most sensitive organs to TCDD exposure. Our experiments revealed a complex distribution of AHR and ARNT mRNAs and proteins in rat and human testis. AHR and ARNT immunoreactivities could be detected in the nuclei of interstitial and tubular cells. The incubation of seminiferous tubules in a serum-free culture medium resulted in up-regulation of AHR mRNA, which could be depressed by adding FSH to the culture medium. Furthermore, the incubation of tubular segments with a solution of 1 or 100 nM TCDD resulted in a 2- to 3-fold increase in apoptotic cells. Thus, up-regulation of AHR in cultured tubular segments and consecutive depression by FSH suggest a role for AHR in controlled cell death during spermatogenesis. We suggest that AHR and ARNT mediate effects by direct action on testicular cells in the rat and human testis. PMID- 12586753 TI - Regulation of iodothyronine deiodinases in the Pax8-/- mouse model of congenital hypothyroidism. AB - Thyroid hormones are essential for a variety of developmental and metabolic processes. Congenital hypothyroidism (CHT) results in severe defects in the development of different tissues, in particular brain. As an animal model for CHT, we studied Pax8(-/-) mice, which are born without a thyroid gland. We determined the expression of iodothyronine deiodinase D1 in liver and kidney, D2 in brain and pituitary, and D3 in brain, as well as serum T(4), T(3), and rT(3) levels in Pax8(-/-) vs. control mice during the first 3 wk of life. In control mice, serum T(4) and T(3) were undetectable on the day of birth (d 0) and increased to maximum levels on d 15. In Pax8(-/-) mice, serum T(4) and T(3) remained below detection limits. Serum rT(3) was high on d 0 in both groups and rapidly decreased in Pax8(-/-), but not in control mice. Hepatic and renal D1 activities and mRNA levels were low on d 0 and increased in control mice roughly parallel to serum T(4) and T(3) levels. In Pax8(-/-) mice, tissue D1 activities and mRNA levels remained low. Cerebral D2 activities were low on d 0 and increased to maximum levels on d 15, which were approximately 10-fold higher in Pax8(-/-) than in control mice. D2 mRNA levels were higher in Pax8(-/-) than in control mice only on d 21. Cerebral D3 activities and mRNA levels were high on d 0 and showed a moderate decrease between d 3 and 15, with values slightly lower in Pax8(-/-) than in control mice. One day after the injection of 200 ng T(4) or 20 ng T(3)/g body weight, tissue deiodinase activities and mRNA levels were at least partially restored toward control levels, with the exception of cerebral D3 activity. In conclusion, these findings show dramatic age and thyroid state dependent changes in the expression of deiodinases in central and peripheral tissues of mice during the first 3 wk of life. PMID- 12586754 TI - Dose-response effects of 2-methoxyestradiol on estrogen target tissues in the ovariectomized rat. AB - In three experiments, we evaluated the pharmacological effects of 2 methoxyestradiol (2ME(2)) on several estrogen target tissues. Experiment 1: we gavaged recently ovariectomized (OVX) 9.5-wk-old rats with 2ME(2) at doses of 0, 0.1, 1, 4, 20, and 75 mg/kg in a 21-d dose-response study. 2ME(2) reduced body weight and serum cholesterol, increased uterine weight and epithelial cell height, and inhibited longitudinal and radial bone growth compared with values in the untreated OVX rat. All doses of 2ME(2) maintained cancellous bone mass at the baseline level, the lowest effective dose being 20-fold less than a uterotrophic dose. Experiment 2: in an 8-wk experiment in adult OVX rats, a nonuterotrophic dose of 2ME(2) (4 mg/kg x d) suppressed body weight gain, inhibited bone formation in cancellous bone and partially prevented bone loss in the tibial metaphysis. Experiment 3: in weanling rats, ICI 182,780 did not antagonize the effect of 2ME(2). We conclude that 2ME(2) antagonizes the skeletal changes that follow OVX at doses that have minimal or no effects in the uterus in both young and adult rats; 2ME(2) does not appear to act via estrogen receptors and is active on bone at doses well below those required for tumor suppression in mice. 2ME(2), through a novel pathway, may be a useful alternative to conventional hormone replacement therapy for prevention of postmenopausal bone loss. PMID- 12586755 TI - Follicle-stimulating hormone and insulin-like growth factor I synergistically induce up-regulation of cartilage link protein (Crtl1) via activation of phosphatidylinositol-dependent kinase/Akt in rat granulosa cells. AB - FSH and IGF-I are both important determinants of follicle development and the process of cumulus cell-oocyte complex expansion. FSH stimulates the phosphorylation of Akt by mechanisms involving phosphatidylinositol 3-kinase (PI3 K), a pattern of response mimicking that of IGF-I. Cartilage link protein (Crtl1) is confined to the cartilaginous lineage and is assembled into a macroaggregate complex essential for hyaluronan-rich matrix stabilization. The present studies were performed to determine the actions of FSH and IGF-I on Crtl1 production in rat granulosa cells. Primary cultures of granulosa cells were prepared from 24-d old rats. After treatments, cell extracts and media were prepared, and the Crtl1 level was determined by immunoblotting analysis using anti-Crtl1 antibodies. Here we showed that 1) treatment with FSH (> or = 25 ng/ml) or IGF-I (> or = 25 ng/ml) for 4 h increased Crtl1 production; 2) maximal stimulatory effects of FSH or IGF I were observed at 100 or 50 ng/ml, respectively; 3) FSH caused a concentration dependent increase in IGF-I-induced Crtl1 production and vice versa; 4) FSH and IGF-I also up-regulate the expression of Crtl1 mRNA; 5) FSH- and IGF-I-dependent Crtl1 production were abrogated by PI3-K inhibitors (LY294002 and wortmannin), and inhibition of Crtl1 production by p38 mitogen-activated protein kinase inhibitor (SB202190) was partial (approximately 30%), suggesting that PI3-K and, to a lesser extent, p38 mitogen-activated protein kinase are critical for the response. Our study represents the first report that FSH amplifies IGF-I-mediated Crtl1 production, possibly via PI3-K-Akt signaling cascades in rat granulosa cells. PMID- 12586756 TI - Characterization of heme oxygenase 1 (heat shock protein 32) induction by atrial natriuretic peptide in human endothelial cells. AB - BACKGROUND: Atrial natriuretic peptide (ANP) is a cardiovascular hormone possessing antiinflammatory and cytoprotective potential. The aim of this study was to characterize induction of heme oxygenase (HO)-1 by ANP in human umbilical vein endothelial cells (HUVEC). METHODS: HUVEC were treated with ANP, 8-bromo cyclic GMP (cGMP), or cANF in the presence or absence of various inhibitors. HO-1 was determined by Western blot and RT-PCR, c-jun N-terminal kinase (JNK) and ERK by the use of phospho-specific antibodies. Activator protein (AP)-1 activation was assessed by gelshift assay. Reporter gene assays were performed using native or mutated AP-1 binding sites of the HO-1 promoter. TNF-alpha-induced cell death was investigated by Hoechst staining, fluorescence-activated cell sorting analysis, caspase-3-measurement, and 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide test. RESULTS: ANP (10(-9)-10(-6) mol/liter) induced the expression of HO-1 protein and mRNA. Induction was mediated via the guanylate cyclase-coupled receptor because 8-Br-cGMP mimicked the effect of ANP, whereas the clearance receptor agonist cANF did not induce HO-1. Endogenously produced cGMP also induced HO-1 because phosphodiesterase inhibition markedly elevated HO 1. The lack of effect of the cGMP-dependent protein kinase inhibitor 8-(4 chlorophenylthio)guanosine-3',5'-cyclic monophosphorothioate, Rp-isomer (Rp-8-pCT cGMPS) suggested no involvement for this cGMP effector pathway in the signal transduction. ANP lead to activation of the transcription factor AP-1, and subsequently of JNK, as well as of ERK. Cotreatment of the cells with U0126 or SP600125, as well as reporter gene assays revealed the involvement of AP-1/JNK activation in HO-1 induction. Abrogation of HO-1 induction by PD-98059 showed also a role for ERK. Treatment of HUVEC with ANP did not protect from TNF-alpha induced apoptosis. CONCLUSION: This work characterizes the induction of HO-1 by ANP in HUVEC, which is shown to be mediated via JNK/AP-1 and ERK pathways. ANP induced HO-1 does not confer protection against TNF-alpha-induced apoptosis. PMID- 12586757 TI - A role for galanin-like peptide in the integration of feeding, body weight regulation, and reproduction in the mouse. AB - Galanin-like peptide (GALP) shares sequence homology with galanin and binds to galanin receptors in vitro. GALP neurons in the arcuate nucleus coexpress leptin receptors, and GALP mRNA expression is up-regulated by leptin. Based on these observations, we postulated that GALP plays a role in mediating leptin's inhibitory effects on food intake (FI) and body weight (BW), as well as its stimulatory effect on the reproductive axis. To test these hypotheses, we performed several studies in which mice received intracerebroventricular injections of either GALP or vehicle. Acute GALP treatment elicited a dose dependent suppression of FI and BW. Long-term treatment with GALP caused only transient reductions in FI and BW, demonstrating that the mice became refractory to continued exposure to GALP. GALP inhibited FI as early as 1 h post injection. Central injection of GALP suppressed locomotor activity and elicited the formation of a conditioned taste aversion. In male mice, serum levels of LH and testosterone were increased by GALP administration. Although we cannot rule out possible nonspecific effects of GALP on FI, the present observations are consistent with the argument that GALP is a downstream effector of leptin's actions within the central nervous system. PMID- 12586758 TI - Gonadotropin-releasing hormone neurons generate interacting rhythms in multiple time domains. AB - Pulsatile release of GnRH is prerequisite for fertility. The possibility that multiple rhythms interact to generate GnRH pulses was raised by observations of changes in action potential firing and intracellular calcium levels occurring much more frequently than hormone pulses. To examine this further, we analyzed firing patterns from targeted extracellular recordings of green fluorescent protein-expressing GnRH neurons in acute brain slices prepared from adult ovariectomized and ovariectomized +estradiol mice. Fourier spectral analysis identified rhythms in multiple time domains, which we grouped into bursts (a period of <100 sec), clusters (100-1000 sec), or episodes (>1000 sec). Bursts were the fundamental unit of activity and consisted of trains of action currents (the currents during action potentials). Episodes and clusters were lower frequency changes in firing rate resulting from alterations in the time between bursts. Specifically, mean interburst interval during episode peaks was less than during nadirs. In contrast, neither burst duration nor action currents/burst differed between peaks and nadirs. Estradiol increased episode period by changing the patterning of bursts, not burst duration or action currents/burst. We propose a low frequency rhythm that is subject to external influences alters the patterning of a fundamental unit of activity to change ultimately GnRH pulse frequency. PMID- 12586759 TI - Estrogen activation of cyclic adenosine 5'-monophosphate response element mediated transcription requires the extracellularly regulated kinase/mitogen activated protein kinase pathway. AB - The ability of estrogen to rapidly initiate a variety of signal transduction cascades is increasingly recognized as playing an important role in a number of tissue-specific transcriptional actions of the hormone. In vivo, estrogen rapidly elicits phosphorylation of cAMP response element-binding protein (CREB). We have previously shown that both ER alpha and ER beta are capable of activating the MAPK pathway in response to a low dose of 17beta-estradiol. In the present study, the ability of estrogen to act through both ER alpha and ER beta to increase CREB phosphorylation was evaluated in an immortalized hippocampal cell line stably expressing either receptor. Estrogen treatment promoted rapid CREB phosphorylation, reaching a maximum by 15 min. This activation is completely blocked by the antiestrogen ICI 182,780, suggesting an estrogen receptor dependent mechanism. The addition of the mitogen/ERK kinase-1 inhibitor, PD98059, also blocked the ability of estrogen to signal to CREB phosphorylation. Estrogen also caused an increase in p90Rsk activity, a critical mediator of MAPK effects. Surprisingly, blockade of the protein kinase A pathway in cells treated with estrogen did not affect estrogen-mediated CREB phosphorylation. Thus, MAPK and p90Rsk appear to be the primary mediators of estrogen-induced gene transcription through ER alpha and ER beta. PMID- 12586760 TI - c-Jun N-terminal kinase activation of activator protein-1 underlies homologous regulation of the gonadotropin-releasing hormone receptor gene in alpha T3-1 cells. AB - Reproductive function is dependent on the interaction between GnRH and its cognate receptor found on gonadotrope cells of the anterior pituitary gland. GnRH activation of the GnRH receptor (GnRHR) is a potent stimulus for increased expression of multiple genes including the gene encoding the GnRHR itself. Thus, homologous regulation of the GnRHR is an important mechanism underlying gonadotrope sensitivity to GnRH. Previously, we have found that GnRH induction of GnRHR gene expression in alpha T3-1 cells is partially mediated by protein kinase C activation of a canonical activator protein-1 (AP-1) element. In contrast, protein kinase A and a cAMP response element-like element have been implicated in mediating the GnRH response of the GnRHR gene using a heterologous cell model (GGH(3)). Herein we find that selective removal of the canonical AP-1 site leads to a loss of GnRH regulation of the GnRHR promoter in transgenic mice. Thus, an intact AP-1 element is necessary for GnRH responsiveness of the GnRHR gene both in vitro and in vivo. Based on in vitro analyses, GnRH appeared to enhance the interaction of JunD, FosB, and c-Fos at the GnRHR AP-1 element. Although enhanced binding of cFos reflected an increase in gene expression, GnRH appeared to regulate both FosB and JunD at a posttranslational level. Neither overexpression of a constitutively active Raf-kinase nor pharmacological blockade of GnRH induced ERK activation eliminated the GnRH response of the GnRHR promoter. GnRH responsiveness was, however, lost in alpha T3-1 cells that stably express a dominant-negative c-Jun N-terminal kinase (JNK) kinase, suggesting a critical role for JNK in mediating GnRH regulation of the GnRHR gene. Consistent with this possibility, we find that the ability of forskolin and membrane-permeable forms of cAMP to inhibit the GnRH response of the GnRHR promoter is associated with a loss of both JNK activation and GnRH-mediated recruitment of the primary AP-1 binding components. PMID- 12586761 TI - Identification of a new growth hormone family protein, somatolactin-like protein, in the rainbow trout (Oncorhyncus mykiss) pituitary gland. AB - A cDNA encoding a new GH family protein expressed in the rainbow trout (Oncorhyncus mykiss) pituitary gland was identified. Because the deduced amino acid sequence of this cDNA shares 56% homology with rainbow trout (rt) somatolactin (rtSL), we named it as rtSL-like protein (rtSLP). Comparison of the amino acid sequences of rtGH, rainbow trout prolactin (rtPRL), and rtSLP revealed that rtSLP shares 26% and 21% identity with that of rtGH and rtPRL, respectively. Trout SLP contains not only the known GH family protein conserved domains but also all of the four cysteine residues that are responsible for the formation of two disulfide linkages in GH, PRL, and SL. Immunoblot analysis revealed that rtSLP is secreted from the pituitary gland once it is synthesized. Unlike rtSL, rtSLP mRNA was readily detectable in pituitary glands of 2-yr fish (30-34 cm in length) but not in embryos or 1-yr fish (17-25 cm in length). Furthermore, the level of rtSLP mRNA in male fish is 2-fold higher than that in female fish. These results suggest that rtSLP might play a role in regulating the reproductive maturation in rainbow trout. PMID- 12586762 TI - Androgen regulation of parathyroid hormone-related peptide production in human prostate cancer cells. AB - PTHrP is the major pathogenetic factor for hypercalcemia in several malignancies including prostate cancer. In the current study, we have assessed the ability of androgens to regulate PTHrP production in androgen-insensitive human prostate cancer cells PC-3 and cells transfected with androgen receptor (PC-3T). Androgen responsiveness caused a marked decrease in PC-3T cell growth, and treatment of these cells with dihydrotestosterone led to inhibition of PTHrP production. These inhibitory effects were readily reversed by androgen receptor antagonist flutamide. To determine the effect of androgens on tumor growth and PTHrP production in vivo, PC-3 and PC-3T cells were injected into the right flank of male BALB/c nu/nu mice. Animals inoculated with PC-3 and PC-3T cells developed palpable tumors at wk 2 and 4, respectively. Inoculation of PC-3T cells into castrated animals resulted in rapid tumor growth in PC-3T tumors, effects that were reversed in PC-3T tumors grown in castrated hosts. Using PTHrP promoter luciferase reporter, a 30% decrease in luciferase activity was seen following treatment with dihydrotestosterone. These results indicate that PC-3 cell growth correlates inversely with androgen sensitivity and directly with PTHrP production in vitro and in vivo, androgens can regulate PTHrP production, and the androgen effect on PTHrP is mediated at least in part by transcriptional regulation via the androgen receptor. PMID- 12586763 TI - Interaction of the growth hormone receptor with cytokine-induced Src homology domain 2 protein in rat adipocytes. AB - GH stimulates the phosphorylation of tyrosine residues in the GH receptor (GHR), Janus kinase 2 (JAK2), and other signaling proteins in a transient manner that subsides within 1 h. To assess the possible roles of cytokine-induced Src homology domain 2 (SH2) (CIS/SOCS) proteins in these transient responses, we studied the expression and disposition of CIS/SOCS proteins in rat adipocytes, a physiological target of GH action. A tyrosine-phosphorylated protein that appears to be the GHR was coprecipitated from extracts of GH-treated adipocytes with alpha-CIS. In contrast, no tyrosine-phosphorylated adipocyte proteins were recovered after immunoprecipitation with alpha-SOCS3, although coprecipitation of GHR with SOCS3 was readily detected in extracts of 3T3-F442A fibroblasts. Interaction of GHR with CIS peaked between 2 and 10 min after adipocytes were treated with GH, when tyrosine phosphorylation of the GHR was maximal. By 60 min after GH, tyrosine phosphorylation of the GHR declined to very low levels, and its interaction with CIS was reduced correspondingly. Proteasome inhibitors prevented the decline in tyrosine-phosphorylated GHR and prolonged interaction of GHR and CIS for at least 1 h. These findings demonstrate the interaction of CIS with the GHR in vivo and suggest that CIS may enhance degradation of the receptor by a proteasomal pathway. PMID- 12586764 TI - The 5'-flanking region of the murine epididymal protein of 17 kilodaltons gene targets transgene expression in the epididymis. AB - A murine epididymal retinoic-acid-binding protein (mE-RABP) is specifically expressed in the mid/distal caput epididymidis and is androgen regulated. The murine epididymal protein of 17 kDa (mEP17) gene, a novel gene homologous to mE RABP, is located within 5 kb of the 5'-flanking region of the mE-RABP gene. In contrast, expression of the mEP17 gene is restricted to the initial segment and regulated by factor(s) contained in testicular fluid. To identify cis-DNA regulatory element(s) involved in the tissue- and region-specific expression of the mEP17 gene in transgenic mice, we have studied the expression of a transgene containing 5.3 kb of the 5'-flanking region of the mEP17 gene (5.3mEP17) linked to chloramphenicol acetyltransferase (CAT) reporter gene. Significant caput epididymidis-specific CAT activity was detected in transgenic mouse lines; and CAT gene expression is restricted to the initial segment, as is the expression of the endogenous mEP17 gene. Ontogenic expression and testicular factor dependency also mimic that of endogenous mEP17 gene. These results suggest that the 5.3mEP17 fragment contains all the information required for spatial and temporal expression in the mouse epididymis. The 5.3mEP17 fragment will be useful to express a foreign gene of interest in the epididymis in an initial segment specific manner. PMID- 12586765 TI - Identification, immunolocalization, regulation, and postnatal development of the lipocalin EP17 (epididymal protein of 17 kilodaltons) in the mouse and rat epididymis. AB - Several lipocalins are present in the mouse epididymis and are thought to play a role in sperm maturation by transporting lipophilic molecules. We have previously reported that two lipocalin genes, mERABP (mouse epididymal retinoic acid binding protein), and mEP17 (mouse epididymal protein of 17 kDa), derived from an ancestral gene, are specifically expressed in the epididymis. In the present study, a polyclonal antibody was raised against a recombinant protein to investigate the presence and the regulation of mEP17. mEP17 was detected in the supranuclear region of the principal cells of the initial segment, the clear cells of the caput epididymidis, and the lumen of the mid/distal caput but not of the distal epididymis. Initial segment and caput tissue extracts were subjected to HPLC separation. After electrophoresis of the immunoreactive mEP17-enriched fractions, the immunoreactive band was analyzed by mass spectrometry to identified mEP17 unambiguously. After two-dimensional electrophoresis, mEP17 appeared as a train of five 22-kDa spots with a range of pI (isoelectric point) from 5.8-6.7. N-glycanase digestion gave rise to a single spot of 17 kDa and pI 6, the predicted mass and pI. During ontogeny, mEP17 was detected as early as 3 wk of age and increased afterward. After bilateral orchiectomy, mEP17 disappeared 2 d after surgery and was not restored after testosterone replacement. After unilateral orchiectomy, mEP17 levels decreased only in the orchiectomized side. After cryptorchidism or busulfan treatment, mEP17 levels were either greatly diminished or not detected. This suggests that mEP17 is dependent on testicular factor(s) that may have a germ cell origin. Altogether, our data demonstrate that mEP17 spatial expression, regulation, and fate are different from that of the highly related mouse epididymal retinoic acid binding protein. This suggests that these two related proteins exhibit distinct functions in the mouse epididymis. PMID- 12586766 TI - The cystatin-related epididymal spermatogenic protein inhibits the serine protease prohormone convertase 2. AB - The cystatin-related epididymal spermatogenic (CRES) protein is related to the family 2 cystatins of the cystatin superfamily of cysteine protease inhibitors. However, CRES lacks sequences important for cysteine protease inhibitory activity and is specifically expressed in reproductive and neuroendocrine tissues. Thus, CRES is distinct from cystatins and may perform unique tissue-specific functions. The purpose of the present study was to determine whether CRES functions as a protease inhibitor in in vitro assays. In contrast to mouse recombinant cystatin C, recombinant CRES did not inhibit the cysteine proteases papain and cathepsin B, suggesting that it probably does not function as a typical cystatin. CRES, however, inhibited the serine protease prohormone convertase 2 (PC2), a protease involved in prohormone processing in the neuroendocrine system, whereas cystatin C showed no inhibition. CRES did not inhibit subtilisin, trypsin, or the convertase family members, PC1 and furin, indicating that it selectively inhibits PC2. Kinetic analysis showed that CRES is a competitive inhibitor of PC2 with a K(i) of 25 nM. The removal of N-terminal sequences from CRES decreased its affinity for PC2, suggesting that the N terminus may be important for CRES to function as an inhibitor. These studies suggest that CRES is a cross-class inhibitor that may regulate proprotein processing within the reproductive and neuroendocrine systems. PMID- 12586767 TI - Cres2 and Cres3: new members of the cystatin-related epididymal spermatogenic subgroup of family 2 cystatins. AB - The cystatin-related epididymal spermatogenic (CRES) and recently identified testatin and cystatin T proteins define a new subgroup within the family 2 cystatins of cysteine protease inhibitors. Members of the CRES subgroup are predominantly expressed in reproductive tissues and lack critical cystatin active site sequences implying divergent functions. To determine whether there are additional members of the subgroup, we searched nucleotide databases and identified two novel genes that we designated Cres2 and Cres3. These genes, like other subgroup members, encode proteins with four conserved cysteine residues and predicted molecular weights characteristic of family 2 cystatins but have divergent cystatin inhibitory sequences. Furthermore, the genes exhibited reproductive-specific expression with Cres2 exclusively expressed in the epithelial cells of the proximal and midcaput epididymal regions and Cres3 expressed in the proximal caput epididymal epithelium, Sertoli cells of the testis, and early follicles and corpora lutea in the ovary. Additional studies showed that, like Cres, both Cres2 and Cres3 genes are dependent on testicular factors for epididymal expression. Taken together, CRES2 and CRES3 represent new members of a subgroup of cystatin family 2 proteins that likely carry out tissue specific functions distinct from that of typical cystatins. PMID- 12586768 TI - Dose-dependent inhibition by ghrelin of insulin secretion in the mouse. AB - Ghrelin is produced by stomach oxyntic cells and thought to be involved in the regulation of body weight and food intake. We demonstrate here that the peptide inhibits insulin secretion from overnight-incubated mouse islets in the presence of 8.3, 11.1, and 22.2 mmol/liter glucose. Ghrelin was most efficient at 1 nmol/liter and its effect disappeared by raising the dose more than 25 nmol/liter. Also, insulin secretion in the presence of high K(+) concentrations (20 mmol/liter) was inhibited by ghrelin. Furthermore, when administered iv to mice together with glucose (1 g/kg), ghrelin (50 nmol/kg) inhibited both the rapid 1-min insulin response (364 +/- 90 vs. 985 +/- 114 pmol/liter in controls, P < 0.001) and the area under the 50 min curve of insulin concentration (12.6 +/- 1.2 vs. 15.6 +/- 1.2 nmol/liter x 50 min; P = 0.046) without affecting the glucose disposal rate, insulin sensitivity or glucose effectiveness, i.e. glucose disposal independent from any dynamic change in insulin. The insulinostatic effect of ghrelin was inversely related to insulin sensitivity. In contrast, ghrelin had no influence at the lower dose of 5 nmol/kg and only slightly inhibited insulin secretion at the higher dose of 150 nmol/kg. These findings therefore show that ghrelin inhibits glucose-stimulated insulin secretion in the mouse. The effect is dependent on the dose and elicited on distal signaling steps in islet cells. The results suggest that the islet beta-cells are targets for ghrelin. PMID- 12586769 TI - The signaling adapters fibroblast growth factor receptor substrate 2 and 3 are activated by the thyroid TRK oncoproteins. AB - The thyroid TRK oncogenes are generated by chromosomal rearrangements juxtaposing the neurotrophic tyrosine receptor kinase type 1 (NTRK1) tyrosine kinase domain to foreign activating sequences. TRK oncoproteins display a constitutive tyrosine kinase activity resulting in the capability to transform NIH3T3 cells. The TRK oncoproteins' signal transduction has been in part elucidated, and it involves several signal transducers activated by the NGF-stimulated NTRK1 receptor. In this paper, we investigate the role of FRS2 and FRS3, two related adapter proteins activated by fibroblast growth factor and NTRK1 receptors, in the signaling of the thyroid TRK-T1 and TRK-T3 oncogenes. By a combination of in vitro and in vivo assays, we demonstrate that both fibroblast growth factor receptor substrate (FRS)2 and FRS3 are recruited and activated by TRK-T1 and TRK T3. Interaction studies using different TRK-T3 mutants indicate that FRS3 is recruited by the same tyrosine residue interacting with Shc and FRS2. Expression studies show different expression patterns of the FRS adapters in normal and tumor thyroid samples: FRS3 is expressed in both normal and thyroid tumor samples, whereas FRS2 is not expressed in normal thyroid but is differentially expressed in some tumors. Altogether, our data indicate that the FRS2 and FRS3 adapters may have a role in thyroid carcinogenesis triggered by TRK oncogenes. PMID- 12586770 TI - Insulin-like growth factor regulates peak bone mineral density in mice by both growth hormone-dependent and -independent mechanisms. AB - To evaluate the relative contribution of the GH/IGF axis to the development of peak bone mineral density (BMD), we measured skeletal changes in IGF-I knockout (KO), IGF-II KO, and GH-deficient lit/lit mice and their corresponding control mice at d 23 (prepubertal), 31 (pubertal), and 56 (postpubertal) in the entire femur by dual energy x-ray absorptiometry and in the mid-diaphysis by peripheral quantitative computed tomography. Lack of growth factors resulted in different degrees of failure of skeletal growth depending on the growth period and the growth factor involved. At d 23, femoral length, size, and BMD were reduced by 25 40%, 15-17%, and 8-10%, respectively, in mice deficient in IGF-I, IGF-II, and GH compared with the control mice. During puberty, BMD increased by 40% in control mice and by 15% in IGF-II KO and GH-deficient mice, whereas it did not increase in the IGF-I KO mice. Disruption of IGF-I, but not IGF-II, completely prevented the periosteal expansion that occurs during puberty, whereas it was reduced by 50% in GH-deficient mice. At d 56, femoral length, size, and BMD were reduced by 40-55%, 11-18%, and 25-32%, respectively, in mice deficient in IGF-I, IGF-II, and GH compared with the control mice. Our data demonstrate that: 1) mice deficient in IGF-I exhibit a greater impairment in bone accretion than mice deficient in IGF-II or GH; 2) GH/IGF-I, but not IGF-II, is critical for puberty-induced bone growth; and 3) IGF-I effects on bone accretion during prepuberty are mediated predominantly via mechanisms independent of GH, whereas during puberty they are mediated via both GH-dependent and GH-independent mechanisms. PMID- 12586771 TI - In vivo dimerization of types 1, 2, and 3 iodothyronine selenodeiodinases. AB - The goal of the present investigation was to test the hypothesis that types 1, 2, and 3 iodothyronine selenodeiodinases (D1, D2, and D3) can form homodimers. The strategy included transient coexpression of wild-type (wt) deiodinases (target), and FLAG-tagged alanine or cysteine mutants (bait) in human embryonic kidney epithelial cells. SDS-PAGE of the immunoprecipitation pellet of (75)Se-labeled cell lysates using anti-FLAG antibody revealed bands of the correct sizes for the respective wt enzymes, which corresponded to approximately 2-5% of the total deiodinase protein in the cell lysate. Western blot analysis with anti-FLAG antibody of lysates of cells transiently expressing individual FLAG-tagged cysteine deiodinases revealed specific monomeric bands for each deiodinase and additional minor bands of relative molecular mass (M(r)) of 55,000 for D1, M(r) 62,000 for D2, and M(r) 65,000 for D3, which were eliminated by 100 mM dithiothreitol at 100 C. Anti-FLAG antibody immunodepleted 10% of D1 and 38% of D2 activity from lysates of cells coexpressing inactive FLAG-tagged Ala mutants and the respective wt enzymes (D1 or D2) but failed to immunodeplete wtD3 activity. D1 or D2 activities were present in these respective pellets. We conclude 1) that overexpressed selenodeiodinases can homodimerize probably through disulfide bridges; and 2) at least for D1 and D2, monomeric forms are catalytically active, demonstrating that only one wt monomer partner is required for catalytic activity of these two deiodinases. PMID- 12586772 TI - Defective activation of atypical protein kinase C zeta and lambda by insulin and phosphatidylinositol-3,4,5-(PO4)(3) in skeletal muscle of rats following high-fat feeding and streptozotocin-induced diabetes. AB - Insulin-stimulated glucose transport in skeletal muscle is thought to be effected at least partly through atypical protein kinase C isoforms (aPKCs) operating downstream of phosphatidylinositol (PI) 3-kinase and 3-phosphoinositide-dependent protein kinase-1 (PDK-1). However, relatively little is known about the activation of aPKCs in physiological conditions or insulin-resistant states. Presently, we studied aPKC activation in vastus lateralis muscles of normal chow fed and high-fat-fed rats and after streptozotocin (STZ)-induced diabetes. In normal chow-fed rats, dose-dependent increases in aPKC activity approached maximal levels after 15-30 min of stimulation by relatively high and lower, presumably more physiological, insulin concentrations, achieved by im insulin or ip glucose administration. Insulin-induced activation of aPKCs was impaired in both high-fat-fed and STZ-diabetic rats, but, surprisingly, IRS-1-dependent and IRS-2-dependent PI 3-kinase activation was not appreciably compromised. Most interestingly, direct in vitro activation of aPKCs by PI-3,4,5-(PO(4))(3), the lipid product of PI 3-kinase, was impaired in both high-fat-fed and STZ-diabetic rats. Defects in activation of aPKCs by insulin and PI-3,4,5-(PO(4))(3) could not be explained by diminished PDK-1-dependent phosphorylation of threonine-410 in the PKC-zeta activation loop, as this phosphorylation was increased even in the absence of insulin treatment in high-fat-fed rats. CONCLUSIONS: 1) muscle aPKCs are activated at relatively low, presumably physiological, as well as higher supraphysiological, insulin concentrations; 2) aPKC activation is defective in muscles of high-fat-fed and STZ-diabetic rats; and 3) defective aPKC activation in these states is at least partly due to impaired responsiveness to PI-3,4,5 (PO(4))(3), apparently at activation steps distal to PDK-1-dependent loop phosphorylation. PMID- 12586773 TI - Opposing roles for the insulin-like growth factor (IGF)-II and mannose 6 phosphate (Man-6-P) binding activities of the IGF-II/Man-6-P receptor in the growth of prostate cancer cells. AB - The IGF-II/mannose 6-phosphate (Man-6-P) receptor (IGF2R) binds IGF-II and Man-6 P-bearing ligands at distinct binding sites. Analysis of IGF2R expression and function suggested that decreased IGF2R expression could partly account for the increased growth of lymph node carcinoma of the prostate (LNCaP) human prostate cancer cells observed with increasing passage in culture. However, LNCaP cells that expressed a Myc-tagged IGF2R (IGF2RMyc) proliferated more rapidly than control cells transfected with the empty vector. LNCaP cells expressing a mutant IGF2R incompetent to bind IGF-II (IGF2RMyc I/T) proliferated more rapidly than both vector-transfected cells and cells expressing the IGF2RMyc. In contrast, forced expression of IGF2RMyc in PC-3 human prostate cancer cells resulted in decreased proliferation, compared with control cells. As in LNCaP cells, PC-3 cells expressing IGF2RMyc I/T proliferated more rapidly than vector-transfected cells. The subcellular distribution and ability to internalize cell-surface IGF II of IGF2RMyc were indistinguishable from endogenous IGF2R in PC-3 cells. These data suggest that the IGF-II- and Man-6-P-binding functions of the IGF2R have opposing activities, with respect to growth of prostate cancer cells. The magnitude of each activity in a given cell type seems to determine whether the net effect of the IGF2R on cell growth is inhibitory or stimulatory. PMID- 12586774 TI - Interrelationship between the novel peptide ghrelin and somatostatin/growth hormone-releasing hormone in regulation of pulsatile growth hormone secretion. AB - GH is an anabolic hormone that is essential for normal linear growth and has important metabolic effects throughout life. The ultradian rhythm of GH secretion is generated by the intricate patterned release of two hypothalamic hormones, somatostatin (SRIF) and GHRH, acting both at the level of the pituitary gland and within the central nervous system. The recent discovery of ghrelin, a novel GH releasing peptide identified as the endogenous ligand for the GH secretagogue receptor and shown to induce a positive energy balance, suggests the existence of an additional neuroendocrine pathway for GH control. To further understand how ghrelin interacts with the classical GHRH/SRIF neuronal system in GH regulation, we used a combined physiological and histochemical approach. Our physiological studies of the effects of ghrelin on spontaneous pulsatile GH secretion in conscious, free-moving male rats demonstrate that 1) ghrelin, administered either systemically or centrally, exerts potent, time-dependent GH-releasing activity under physiological conditions; 2) ghrelin is a functional antagonist of SRIF, but its GH-releasing activity at the pituitary level is not dependent on inhibiting endogenous SRIF release; 3) SRIF antagonizes the action of ghrelin at the level of the pituitary gland; and 4) the GH response to ghrelin in vivo requires an intact endogenous GHRH system. Our dual chromogenic and autoradiographic in situ hybridization experiments provide anatomical evidence that ghrelin may directly modulate GHRH mRNA- and neuropeptide Y mRNA-containing neurons in the hypothalamic arcuate nucleus, but that SRIF mRNA-expressing cells are not major direct targets for ghrelin. Together, these findings support the idea that ghrelin may be a critical hormonal signal of nutritional status to the GH neuroendocrine axis serving to integrate energy balance and the growth process. PMID- 12586775 TI - Gene expression is differentially regulated in the epididymis after orchidectomy. AB - The epididymis is the site for the transport, maturation, and storage of spermatozoa. Regulation of epididymal structure and function is highly dependent on the ipsilateral testis. At the molecular level, however, few studies have been undertaken to determine which genes are expressed in the epididymis under testicular regulation. The goal of this study was to identify genes for which expression is regulated after orchidectomy, both throughout the epididymis and in a segment-specific manner. Microarrays spotted with 474 rat cDNAs were used to examine gene expression changes over the first 7 d post orchidectomy in the initial segment, caput, corpus, and cauda epididymidis of the adult Brown Norway rat. Using k-means cluster analysis, we show that four patterns of gene expression are activated in each epididymal segment over the first week following orchidectomy. Transient up-regulation of gene expression in the epididymis after orchidectomy is described for the first time. Potential androgen-repressed genes, including Gpx-1, show increased expression in the epididymis after orchidectomy. Several glutathione-S-transferases and calcium-binding proteins decline throughout the epididymis after orchidectomy, indicating that these may be novel androgen-regulated epididymal genes. Other genes coding for metabolism-associated proteins, transporters, and alpha-1 acid glycoprotein show segment-specific regulation in the epididymis after orchidectomy. Finally, we describe the expression of the previously uncharacterized heat shock proteins, and apoptosis associated genes in the epididymis after orchidectomy. Thus, gene expression in the epididymis is differentially affected over time after orchidectomy. These results provide novel insight into androgen-dependent and segment-specific epididymal function. PMID- 12586776 TI - Contrasting effects of prenyltransferase inhibitors on estrogen-dependent cell cycle progression and estrogen receptor-mediated transcriptional activity in MCF 7 cells. AB - Activation of estrogen receptors (ERs) by estrogens triggers both ER nuclear transcriptional activity and Src/Ras/Erks pathway-dependent mitogenic activity. The present study implicates prenylated proteins in both estrogenic actions. The farnesyltransferase and geranylgeranyltransferase I inhibitors (FTI-277 and GGTI 298, respectively) antagonize estradiol-stimulated cell cycle progression, progesterone receptor, cyclin D1, and c-Myc expression. In contrast, the inhibitors markedly stimulate transcription from two genes containing estrogen response elements, both in the absence and presence of estradiol. The pure antiestrogen ICI 182,780 inhibits by more than 85% these effects on transcription. We demonstrate that both FTI-277 and GGTI-298 increase the association of steroid receptor coactivator-1 with ER alpha and FTI-277 decreases the association of ER alpha with the histone deacetylase 1, a known transcriptional repressor. In addition, FTI-277 has no marked effect on the association of the two corepressors, nuclear receptor corepressor and silencing mediator of retinoid and thyroid receptor with ER alpha, whereas GGTI-298, similar to tamoxifen, clearly increased these associations. Together, these results demonstrate that prenylated proteins play a role in estradiol stimulation of proliferation and progesterone receptor expression. However, they antagonize the ability of ER alpha to stimulate estrogen response element-dependent transcriptional activity, acting presumably through coregulator complex formation. PMID- 12586777 TI - Programming of rat muscle and fat metabolism by in utero overexposure to glucocorticoids. AB - In utero overexposure to glucocorticoids may explain the association between low birth weight and subsequent development of the metabolic syndrome. We previously showed that prenatal dexamethasone (dex) exposure in the rat lowers birth weight and programs adult fasting and postprandial hyperglycemia, associated with increased hepatic gluconeogenesis driven by elevated liver glucocorticoid receptor (GR) expression. This study aimed to determine whether prenatal dex (100 microg/kg per day from embryonic d 15 to embryonic d 21) programs adult GR expression in skeletal muscle and/or adipose tissue and whether this contributes to altered peripheral glucose uptake or metabolism. In utero dex-exposed rats remained lighter until 6 months of age, despite some early catch-up growth. Adults had smaller epididymal fat pads, with a relative increase in muscle size. Although glycogen storage was reduced in quadriceps, 2-deoxyglucose uptake into extensor digitorum longus muscle was increased by 32% (P < 0.05), whereas uptake in other muscles and adipose beds was unaffected by prenatal dex. GR mRNA was not different in most muscles but selectively reduced in soleus (by 23%, P < 0.05). However, GR mRNA was markedly increased specifically in retroperitoneal fat (by 50%, P < 0.02). This was accompanied by a shift from peroxisomal proliferator activated receptor gamma 1 to gamma 2 expression and a reduction in lipoprotein lipase mRNA (by 28%, P < 0.02). Adipose leptin, uncoupling protein-3 and resistin mRNAs, muscle GLUT-4, and circulating lipids were not affected by prenatal dex. These data suggest that hyperglycemia in 6-month-old rats exposed to dexamethasone in utero is not due to attenuated peripheral glucose disposal. However, increased GR and attenuated fatty acid uptake specifically in visceral adipose are consistent with insulin resistance in this crucial metabolic depot and could indirectly contribute to increased hepatic glucose output. PMID- 12586778 TI - Decreased expression of tumor necrosis factor-alpha-stimulated gene 6 in cumulus cells of the cyclooxygenase-2 and EP2 null mice. AB - Ovulation, the release of fertilizable oocytes from mature follicles, involves tissue remodeling and increased prostaglandin (PG) signaling. Cyclooxygenase (COX)-2 is the rate-limiting enzyme during PG synthesis. Female mice null for either COX-2 or the PGE(2) receptor EP2 are infertile, show decreased ovulation, and exhibit abnormal cumulus expansion. Cumulus expansion is the production of a complex extracellular matrix surrounding the cumulus-oocyte complex (COC). Matrix components consist of hyaluronan, proteoglycans, and proteins with hyaluronan binding domains. One such hyaluronan binding protein is TNFalpha-stimulated gene 6 (TSG-6). By various methods, we show induction of TSG-6 and hyaluronan synthase 2 mRNA in ovaries of mice treated with pregnant mare serum gonadotropin and human chorionic gonadotropin. By in situ hybridization, we show that both genes are expressed in periantral mural granulosa cells and cumulus cells of the mouse ovary. Notably, RT-PCR and in situ hybridization show that TSG-6 mRNA but not hyaluronan synthase-2 mRNA expression is selectively reduced in cumulus cells of COX-2 and EP2 null mice. Western analysis further confirms that TSG-6 protein is reduced in isolated COCs but remains covalently associated with inter alpha trypsin inhibitor in COX-2 null mice. These observations identify TSG-6 as a target of PG action and show that its production in ovulatory follicles is associated with proper formation of the cumulus-derived extracellular matrix. PMID- 12586779 TI - Hormone-regulated expression and localization of versican in the rodent ovary. AB - During ovulation, production of a specialized hyaluronan (HA)-rich matrix cross linked by associated HA binding factors causes expansion of the cumulus oocyte complex. Versican is a member of the hyalectan family that binds HA, provides structure and elasticity to tissues, and impacts cell motility and adhesion. In these studies, we sought to determine whether versican is synthesized by ovulating follicles and localizes along with HA in the expanded cumulus oocyte complex matrix in rodent ovaries and whether its expression and/or localization is altered in anovulatory mutant mice. Analysis of mRNA and protein identified isoforms V0, V1, and V3 versican in mouse and rat ovaries throughout follicular development. In situ hybridization localized versican mRNA most specifically to the granulosa cells. Expression was not significantly altered by estradiol or FSH treatment but was increased up to 10-fold during the periovulatory period after human chorionic gonadotropin treatment. In cultured granulosa cells, forskolin and phorbol 12 myristate 13-acetate or FSH + testosterone increased expression of versican. Immunohistochemical analyses verified versican protein in ovulating follicles localized to the expanded cumulus matrix as well as adjacent to the basement membrane. After ovulation, versican was localized around newly formed corpora lutea and vasculature. Unexpectedly, immunohistochemical analyses also demonstrated versican protein on granulosa cells in early primary and small antral follicles. Versican expression and localization were not altered in progesterone receptor or cyclooxygenase-2 null mice, suggesting that transcription of the versican gene is not a target of these two ovulatory mediators. These observations suggest that versican (V0, V1, and V3) is a matrix component of the granulosa layer throughout folliculogenesis and is enriched in remodeling matrices during ovulation and neovascularization of the corpora lutea. PMID- 12586780 TI - Elevated levels of epidermal growth factor receptor/c-erbB2 heterodimers mediate an autocrine growth regulatory pathway in tamoxifen-resistant MCF-7 cells. AB - The development of acquired resistance to antihormonal agents in breast cancer is a major therapeutic problem. We have developed a tamoxifen-resistant (TAM-R) MCF 7 breast cancer cell line to investigate the mechanisms behind this condition. Both epidermal growth factor receptor (EGFR) and c-erbB2 mRNA and protein expression were increased in TAM-R compared with wild-type MCF-7 cells, whereas comparable levels of c-erbB3 mRNA and protein were expressed in both cell lines. Under basal conditions, phosphorylated EGFR/c-erbB2, EGFR/c-erbB3 but not c erbB2/c-erbB3 receptor heterodimers were detected in TAM-R cells in association with increased levels of phosphorylated extracellular-signal regulated kinase 1/2 (ERK1/2). Both cell lines were capable of generating a range of EGFR-specific ligands and increased expression of transforming growth factor alpha was observed in TAM-R cells. Treatment of TAM-R cells with ZD1839 (Iressa) or trastuzumab (Herceptin) blocked c-erbB receptor heterodimer formation and phosphorylation, reduced ERK1/2 activity, and strongly inhibited cell growth. The MAPK kinase inhibitor PD098059 specifically reduced phosphorylated ERK1/2 levels and inhibited TAM-R growth. All three agents abolished ERK1/2 activity in wild-type cells but caused only small reductions in cell proliferation. These results demonstrate that TAM-R MCF-7 cell growth is mediated by the autocrine release and action of an EGFR-specific ligand inducing preferential EGFR/c-erbB2 dimerization and downstream activation of the ERK pathway. PMID- 12586781 TI - Divergent expression of type 2 deiodinase and the putative thyroxine-binding protein p29, in rat brain, suggests that they are functionally unrelated proteins. AB - Deiodinases (D1, D2, and D3) are selenoproteins involved in thyroid hormone metabolism. Generation of the active hormone T(3), from T(4), is carried out by D1 and D2, whereas D3 degrades both hormones. The identity of the cloned D2 as a selenoprotein is well supported by biochemical and physiological data. However, an alternative view has proposed that type 2 deiodinase is a nonselenoprotein complex containing a putative T(4) binding subunit called p29, with an almost identity in sequence with the Dickkopf protein Dkk3. To explore a possible functional relationship between p29 and D2, we have compared their mRNA expression patterns in the rat brain. In brain, parenchyma p29 was expressed in neurons. High expression levels were found in all the regions of the blood cerebrospinal fluid (CSF) barrier. p29 was present in different types of cells than D2, with the exception of the tanycytes. Our data do not support that p29 has a functional relationship with D2. On the other hand, expression of p29 in the blood-CSF barrier suggests that it might be involved in T(4) transport to and from the CSF, but further studies are needed to substantiate this hypothesis. PMID- 12586782 TI - Midgestational lethality in mice lacking the parathyroid hormone (PTH)/PTH related peptide receptor is associated with abrupt cardiomyocyte death. AB - PTHrP is a key developmental regulatory protein and a potent vasoactive agent. Previous studies have shown that mice lacking either the Pthrp or the PTH type 1 receptor (Pth1r) gene exhibit severe chondrodysplasia. In addition, in most genetic backgrounds, the receptor null mice die prenatally at midgestation, but the cause of death remains elusive. Here we show the loss of the Pth1r gene in C57BL6 mice leads to massive, abrupt cardiomyocyte death and embryonic lethality between embryonic days (E) E11.5 and E12.5. PTH1R mRNA was abundantly expressed in the developing wild-type mouse heart and cardiomyocytes from E11.5 embryos demonstrated acute increases in cAMP and increased Ca(2+)oscillations in response to PTHrP-(1-34)NH(2). Analyses of more than 300 embryos (E8-E14.5) from C57BL6/PTH1R +/- matings showed that PTH1R-/- mice survived until E11 with no obvious defects in any tissue. By E12, only 10% of the PTH1R-/- embryos survived and all PTH1R null mice were dead by E13. Ultrastructural and histological analysis revealed striking mitochondrial abnormalities at E11.5 and precipitous cardiomyocyte death between E12.0 and E12.5, followed by degenerative changes in the liver and massive necrosis of other tissues. No abnormalities were observed in the yolk sac or placenta implicating the heart degeneration as the primary cause of death. Taken together, these findings indicate that the PTH1R is required for the development of normal cardiomyocyte function. PMID- 12586783 TI - Evidence for a role of the adenosine 5'-triphosphate-binding cassette transporter A1 in the externalization of annexin I from pituitary folliculo-stellate cells. AB - Annexin 1 (ANXA1) has a well-demonstrated role in early delayed inhibitory feedback of glucocorticoids in the pituitary. ANXA1 is located in folliculo stellate (FS) cells, and glucocorticoids act on these cells to externalize and stimulate the synthesis of ANXA1. However, ANXA1 lacks a signal sequence so the mechanism by which ANXA1 is externalized from FS cells was unknown and has been investigated. The ATP-binding cassette (ABC) transporters are a large group of transporters with varied roles that include the externalization of proteins. Glucocorticoid-induced externalization of ANXA1 from an FS cell line (TtT/GF) and rat anterior pituitary was blocked by glyburide, which inhibits ABC transporters. Glyburide also blocked the glucocorticoid inhibition of forskolin-stimulated ACTH release from pituitary tissue in vitro. RT-PCR revealed mRNA and Western blotting demonstrated protein for the ATP binding cassette A1 (ABCA1) transporter in mouse FS, TtT/GF, and A549 lung adenocarcinoma cells from which glucocorticoids also induce externalization of ANXA1. In TtT/GF cells, immunofluorescence labeling revealed a near total colocalization of cell surface ANXA1 and ABCA1. We conclude that ANXA1, which mediates the early delayed feedback of glucocorticoids in the anterior pituitary, is externalized from FS cells by an ABC transporter and that the ABCA1 transporter is a likely candidate. PMID- 12586784 TI - The porcine sodium/iodide symporter gene exhibits an uncommon expression pattern related to the use of alternative splice sites not present in the human or murine species. AB - The sodium/iodide symporter (NIS) is a membrane protein mediating the active transport of iodide into the thyroid gland. NIS, expressed by human, rat, and mouse thyrocytes, is encoded by a single transcript. We identified NIS mRNA species of 3.5 and 3 kb in porcine thyrocytes. Because porcine thyrocytes in primary culture is a widely used experimental system for thyroid iodide metabolism, we further examined the origin and the function of the porcine NIS (pNIS) transcripts. We generated a porcine thyroid cDNA library from which four different clones, pNIS-D, F, J, and Delta J were isolated. pNIS-D encodes a protein of 643 amino acids highly homologous to the human, rat, and mouse NIS. pNIS-F and J differ from each other and from pNIS-D in their C-terminal part. pNIS-Delta J lacks a six-amino-acid segment within the putative transmembrane domain 10. Transiently expressed in Cos-7 cells, the four pNIS-cDNAs led to the synthesis of proteins targeted at the plasma membrane and conferred perchlorate sensitive iodide uptake activities to Cos-7 cells, except pNIS-Delta J, which was devoid of activity. PNIS-D probably derives from the 3.5-kb transcript and pNIS F, J, and Delta J from the 3-kb transcript. The relative abundance of pNIS-D, F, and J transcripts in porcine thyrocytes was about 60%, 35%, and 5%, respectively; the Delta J transcript was not present in detectable amount. By comparing porcine NIS genomic and cDNA sequences, splice donor and acceptor sites accounting for the generation of pNIS-F, J, and Delta J transcripts were identified. None of the combinations of alternative splice sites found in the pig was present in the human, rat or mouse NIS gene. Our data show that porcine NIS gene, contrary to the NIS gene from other species, gives rise to splice variants leading to three active and one inactive NIS proteins. PMID- 12586785 TI - Growth hormone action on proliferation and differentiation of cerebral cortical cells from fetal rat. AB - To define the role of GH during central nervous system development, we performed studies in cultured rat cerebral cortical cells from 14- (E14) and 17-d-old embryos (E17). The expression of GH receptor, IGF-I receptor, and IGF-I mRNAs was confirmed. In E17, GH increased total cell number (3.9-fold), [(3)H]-thymidine incorporation (3.5-fold), proliferating cell nuclear antigen levels (2.5-fold), and bromodeoxyuridine (BrdU)-positive cells (2.5-fold). GH action on nestin/BrdU positive cells was increased in E14 cells at 3 d in vitro (80-fold) but not at 7 d in vitro. In E14 cells, GH increased (9.5-fold) beta-tubulin/BrdU cells. In E17 cells, GH induced neuronal differentiation, as indicated by the absence of beta tubulin/BrdU-positive cells and the 5.9-fold increment of beta-tubulin protein, and increased glial fibrillary acidic protein/BrdU-positive cells (2.5-fold) and glial fibrillary acidic protein expression (4.5-fold). GH-induced proliferation and differentiation was blocked by IGF-I antiserum. GH increased IGF-binding protein-3 (IGFBP-3), IGF-I receptor protein and its phosphorylation. This study shows that GH promotes proliferation of neural precursors, neurogenesis, and gliogenesis during brain development. These responses are mediated by locally produced IGF-I. GH-induced IGFBP-3 may also have a role in these responses. Therefore, GH is able to activate the IGF-I/IGFBP-3 system in these cerebral cells and induce a physiological action of IGF-I. PMID- 12586786 TI - Inducible nitric oxide synthase mediates bone loss in ovariectomized mice. AB - Several clinical studies have shown that bone loss may be attributed to osteoclast recruitment induced by mediators of inflammation. In different experimental paradigms we have recently demonstrated that estrogen exhibits antiinflammatory activity by preventing the induction of inducible nitric oxide synthase (iNOS) and other components of the inflammatory reaction. To verify whether this could explain the estrogen-dependent blockade of osteoporosis, we investigated the effect of ovariectomy in mice in which iNOS activity had been blunted by genetic or pharmacological manipulation. The consequences of iNOS blockade were evaluated initially on bone formation and resorption by histomorphometric analysis. The proximal tibiae of mice with iNOS genotypes revealed that 32 d after ovariectomy bone volume and bone formation rate were significantly decreased, and osteoclast surface was increased. Conversely, in iNOS knockout (iNOSKO) and wild-type (WT) mice treated with a specific inhibitor of iNOS, N-iminoethyl-L-lysine, ovariectomy did not result in bone depletion. In WT mice, ovariectomy also affected bone formation, as shown by a decreased mineral apposition rate. Also in this case, iNOS inactivation prevented the effect of ovariectomy. Immunocytochemical analysis showed that after ovariectomy iNOS protein accumulates in chondrocytes, and a significant increase in nitrotyrosine and poly(ADP-ribose) synthetase staining was observed in the femur metaphyses. The increase in nitrotyrosine and poly(ADP-ribose) synthetase formation induced by ovariectomy was significantly reduced in sections from iNOSKO mice. These data indicate that in WT mice the observed induction of iNOS has functional relevance, because it leads to overproduction of nitric oxide and accumulation of highly reactive molecules, triggering a local inflammatory reaction. These inflammatory foci attract cytokines, well known actors in the mechanism of osteoclastogenesis. In iNOSKO mice the measurements of IL-1 beta, IL 6, and TNFalpha plasma levels showed that ovariectomy fails to elicit the increase observed in WT animals and suggests that iNOS plays a primary role in the protective effects of estrogens. To further support this hypothesis, we show that estradiol-dependent activation of estrogen receptor-alpha blocks phorbol 12 acetate 13-myristate-induced transcription of iNOS promoter in transfected cells, thus demonstrating that the promoter of iNOS is under estrogen negative control. Our findings point to a key role of iNOS in mediating the negative effects of estrogen depletion on bones and provide a novel mechanistic explanation for the effects of menopause in osteoporosis and possibly also in other diseases in which the inflammatory component is elevated. PMID- 12586787 TI - The central role of human chorionic gonadotropin in the formation of human placental syncytium. AB - A number of trophoblast products, including human chorionic gonadotropin (hCG), can increase the formation of human placental syncytium through the differentiation of mononuclear cytotrophoblasts. The present study investigated the central role of hCG in this process by using antisense receptor phosphorothioate oligodeoxynucleotides (ODNs). Culturing cytotrophoblasts with the hCG/LH receptor antisense, but not sense, ODN resulted in a significant decrease in receptor protein levels and inhibited spontaneous as well as exogenous hCG induced increase in differentiation. The hCG/LH receptor antisense ODN also inhibited epidermal growth factor (EGF), TGF-alpha, leukemia inhibitory factor (LIF), but not 8-bromo-cAMP, induced increases in differentiation, suggesting that hCG is required for EGF, TGF-alpha and LIF, but not for the cAMP actions. Although antisense EGF receptor and LIF receptor ODNs inhibited EGF and LIF induced increase in differentiation, respectively, they were ineffective against hCG, suggesting that they use separate pathways, but they both converge on a common pathway requiring the hCG actions. Mechanism of action studies revealed that EGF treatment activates its receptors and MAPK, both of which are required for EGF to increase the differentiation, cAMP levels and activate protein kinase A. In summary, our results demonstrate that hCG is an autocrine and paracrine regulator that is required for EGF, TGF-alpha, and LIF, but not for cAMP to increase human placental syncytium formation. Direct activation of protein kinase A seems to bypass the hCG pathway, perhaps by targeting genes associated with the differentiation. PMID- 12586788 TI - Early inflammatory responses in experimental cardiac hypertrophy and fibrosis: effects of 11 beta-hydroxysteroid dehydrogenase inactivation. AB - In epithelial tissues such as kidney, mineralocorticoid receptors (MR) are protected against glucocorticoid occupancy by the enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta HSD) type 2. If the enzyme is congenitally inactive, or blocked by carbenoxolone, physiologic glucocorticoids act as MR agonists in such tissues. In most nonepithelial tissues, including cardiomyocytes, 11 beta HSD2 is expressed at minimal levels; in these tissues physiologic glucocorticoids act as MR antagonists, with the basis for this tissue selectivity currently unknown. Vascular smooth muscle cells (VSMC) express MR and 11 beta HSD1/2, with 11 beta HSD1 reported to show uncharacteristic oxidase activity, so that VSMC thus constitute a potential physiologic aldosterone target tissue. Because mineralocorticoid/salt administration triggers marked inflammatory responses in coronary vasculature, we reasoned that VSMC (like epithelial) MR may be activated by glucocorticoids if the protective enzyme is blocked. We thus gave uninephrectomized rats 0.9% NaCl solution to drink, and deoxycorticosterone (DOC, as a single 20 mg sc dose) or carbenoxolone (CBX, 2.5 mg/d in the drinking solution). Both DOC and CBX increased systolic blood pressure, heart, and kidney weight, and expression of cyclooxygenase 2, ED-1-positive macrophages, and osteopontin, with effects of both DOC and CBX blocked by the selective MR antagonist eplerenone. These findings suggest that local glucocorticoid excess, reflecting lower VSMC 11 beta HSD1/2 activity may mimic systemic mineralocorticoid excess, and play a direct etiologic role in coronary vascular inflammatory responses under circumstances of a high salt intake. PMID- 12586789 TI - Sequential single-agent chemotherapy for metastatic breast cancer: therapeutic nihilism or realism? PMID- 12586790 TI - Combination chemotherapy for metastatic breast cancer: reaching for the cure. PMID- 12586791 TI - Advanced Hodgkin's disease: ABVD is better, yet is not good enough! PMID- 12586792 TI - How can we tell when cancer vaccines vaccinate? PMID- 12586793 TI - Phase III trial of doxorubicin, paclitaxel, and the combination of doxorubicin and paclitaxel as front-line chemotherapy for metastatic breast cancer: an intergroup trial (E1193). AB - PURPOSE: Between February 1993 and September 1995, 739 patients with metastatic breast cancer were entered on an Intergroup trial (E1193) comparing doxorubicin (60 mg/m(2)), paclitaxel (175 mg/m(2)/24 h), and the combination of doxorubicin and paclitaxel (AT, 50 mg/m(2) and 150 mg/m(2)/24 h, plus granulocyte colony stimulating factor 5 mg/kg) as first-line therapy. Patients receiving single agent doxorubicin or paclitaxel were crossed over to the other agent at time of progression. PATIENTS AND METHODS: Patients were well balanced for on-study characteristics. RESULTS: Responses (complete response and partial response) were seen in 36% of doxorubicin, 34% of paclitaxel, and 47% of AT patients (P =.84 for doxorubicin v paclitaxel, P =.007 for v AT, P =.004 for paclitaxel v AT). Median time to treatment failure (TTF) is 5.8, 6.0, and 8.0 months for doxorubicin, paclitaxel, and AT, respectively (P =.68 for doxorubicin v paclitaxel, P =.003 for doxorubicin v AT, P =.009 for paclitaxel v AT). Median survivals are 18.9 months for patients taking doxorubicin, 22.2 months for patients taking paclitaxel, and 22.0 months for patients taking AT (P = not significant). Responses were seen in 20% of patients crossing from doxorubicin --> paclitaxel and 22% of patients crossing from paclitaxel --> doxorubicin (P = not significant). Changes in global quality-of-life measurements from on-study to week 16 were similar in all three groups. CONCLUSION: (1) doxorubicin and paclitaxel, in the doses used here, have equivalent activity; (2) the combination of AT results in superior overall response rates and time to TTF; and (3) despite these results, combination therapy with AT did not improve either survival or quality of life compared to sequential single-agent therapy. PMID- 12586794 TI - Application of breast cancer risk prediction models in clinical practice. AB - Breast cancer risk assessment provides an estimation of disease risk that can be used to guide management for women at all levels of risk. In addition, the likelihood that breast cancer risk is due to specific genetic susceptibility (such as BRCA1 or BRCA2 mutations) can be determined. Recent developments have reinforced the clinical importance of breast cancer risk assessment. Tamoxifen chemoprevention as well as prevention studies such as the Study of Tamoxifen and Raloxifene are available to women at increased risk of developing breast cancer. In addition, specific management strategies are now defined for BRCA1 and BRCA2 mutation carriers. Risk may be assessed as the likelihood of developing breast cancer (using risk assessment models) or as the likelihood of detecting a BRCA1 or BRCA2 mutation (using prior probability models). Each of the models has advantages and disadvantages, and all need to be interpreted in context. We review available risk assessment tools and discuss their application. As illustrated by clinical examples, optimal counseling may require the use of several models, as well as clinical judgment, to provide the most accurate and useful information to women and their families. PMID- 12586795 TI - Nonadherence to adjuvant tamoxifen therapy in women with primary breast cancer. AB - PURPOSE: Although clinical trials have clearly demonstrated the benefits of tamoxifen in women with primary breast cancer, little is known about how this drug is actually used in the general population. We sought to estimate adherence and predictors of nonadherence in women starting tamoxifen as adjuvant breast cancer therapy. PATIENTS AND METHODS: Subjects were age 18 years or older initiating tamoxifen for primary breast cancer and enrolled in New Jersey's Medicaid or Pharmaceutical Assistance to the Aged and Disabled programs during the study period, from 1990 to 1996 (N = 2,378). Main outcome measures were number of days covered by filled prescriptions for tamoxifen in the first year of therapy with the 4 years after tamoxifen initiation for a subset; predictors of good versus poor adherence. RESULTS: Twenty-three percent of patients missed taking tamoxifen on more than one fifth of days studied, although on average, patients filled prescriptions for tamoxifen for 87% of their first year of treatment. The youngest, oldest, nonwhite, and mastectomy patients had significantly lower rates of adherence; patients who had seen an oncologist before taking tamoxifen had significantly higher rates of adherence. Overall adherence decreased to 50% by year 4 of therapy. CONCLUSION: The mean level of adherence to tamoxifen is high compared with other chronic medications. However, nearly one fourth of patients may be at risk for inadequate clinical response because of poor adherence. Because of the efficacy of tamoxifen therapy in preventing recurrence and death in women with early-stage breast cancer, further efforts are necessary to identify and prevent suboptimal adherence. PMID- 12586796 TI - Randomized comparison of ABVD and MOPP/ABV hybrid for the treatment of advanced Hodgkin's disease: report of an intergroup trial. AB - PURPOSE: In a series of trials, doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) and mechlorethamine, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, and vinblastine (MOPP/ABV) have been identified as effective treatments for Hodgkin's disease. We compared these regimens as initial chemotherapy for Hodgkin's disease. PATIENTS AND METHODS: Adult patients (N = 856) with advanced Hodgkin's disease were randomly assigned to treatment with ABVD or MOPP/ABV. The major end points were failure-free and overall survival, life-threatening acute toxicities, and serious long-term toxicities, including cardiomyopathy, pulmonary toxicity, myelodysplastic syndromes (MDS), and secondary malignancies. RESULTS: The rates of complete remission (76% v 80%, P =.16), failure-free survival at 5 years (63% v 66%, P =.42), and overall survival at 5 years (82% v 81%, P =.82) were similar for ABVD and MOPP/ABV, respectively. Clinically significant acute pulmonary and hematologic toxicity were more common with MOPP/ABV (P =.060 and.001, respectively). There was no difference in cardiac toxicity. There were 24 deaths attributed to initial treatment: nine with ABVD and 15 with MOPP/ABV (P =.057). There have been 18 second malignancies associated with ABVD and 28 associated with MOPP/ABV (P =.13). Thirteen patients have developed MDS or acute leukemia: 11 were initially treated with MOPP/ABV, and two were initially treated with ABVD but subsequently received MOPP-containing regimens and radiotherapy before developing leukemia (P =.011). CONCLUSION: ABVD and the MOPP/ABV hybrid are effective therapies for Hodgkin's disease. MOPP/ABV is associated with a greater incidence of acute toxicity, MDS, and leukemia. ABVD should be considered the standard regimen for treatment of advanced Hodgkin's disease. PMID- 12586797 TI - Interleukin-2 after autologous stem-cell transplantation for adult patients with acute myeloid leukemia in first complete remission. AB - PURPOSE: To determine the disease-free survival (DFS) and toxicity of administering interleukin-2 (IL-2) immunotherapy early after autologous stem-cell transplantation (ASCT) to simulate a graft versus leukemia effect observed in allogeneic transplantation. PATIENTS AND METHODS: Fifty-six patients with acute myeloid leukemia in first remission received a single consolidation of high-dose cytarabine-idarubicin at a median of 1.1 month postremission with the intent to proceed to ASCT and IL-2 9 x 10(6) U/m(2)/24 h for 4 days, followed by 10 days of IL-2 1.6 x 10(6) U/m(2)/24 h on hematologic recovery. RESULTS: Eighty-four percent of patients received the intended ASCT, and 68% of patients received IL-2 treatment. With a median follow-up of 39.4 months (range, 1.2 to 76.3 months), the 2-year cumulative probability of DFS for all 56 patients is 68% (95% confidence interval [CI], 55% to 80%) and 74% (95% CI, 57% to 85%) for the 39 patients undergoing IL-2 treatment after ASCT. The 2-year cumulative probability of DFS for favorable, intermediate, and unfavorable cytogenetics is 88% (95% CI, 59% to 97%), 48% (95% CI, 26% to 67%), and 70% (95% CI, 23% to 93%), respectively. Toxicities from IL-2 were mainly thrombocytopenia, leukopenia, fever, and fluid retention. Two septic deaths occurred during neutropenia, which includes one during consolidation and one during transplant, for an overall 4% mortality rate. CONCLUSION: These results suggest that a moderate dose of IL-2 after high-dose cytarabine-idarubicin-mobilized ASCT is associated with a low regimen-related toxicity and may improve DFS. A phase III study of IL-2 is now warranted. PMID- 12586799 TI - Phase III study of concurrent chemoradiotherapy versus radiotherapy alone for advanced nasopharyngeal carcinoma: positive effect on overall and progression free survival. AB - PURPOSE: Nasopharyngeal carcinoma (NPC) is a radiosensitive and chemosensitive tumor. This randomized phase III trial compared concurrent chemoradiotherapy (CCRT) versus radiotherapy (RT) alone in patients with advanced NPC. PATIENTS AND METHODS: From December 1993 to April 1999, 284 patients with 1992 American Joint Committee on Cancer stage III to IV (M0) NPC were randomly allocated into two arms. Similar dosage and fractionation of RT was administered in both arms. The investigational arm received two cycles of concurrent chemotherapy with cisplatin 20 mg/m(2)/d plus fluorouracil 400 mg/m(2)/d by 96-hour continuous infusion during the weeks 1 and 5 of RT. Survival analysis was estimated by the Kaplan Meier method and compared by the log-rank test. RESULTS: Baseline patient characteristics were comparable in both arms. After a median follow-up of 65 months, 26.2% (37 of 141) and 46.2% (66 of 143) of patients developed tumor relapse in the CCRT and RT-alone groups, respectively. The 5-year overall survival rates were 72.3% for the CCRT arm and 54.2% for the RT-only arm (P =.0022). The 5-year progression-free survival rates were 71.6% for the CCRT group compared with 53.0% for the RT-only group (P =.0012). Although significantly more toxicity was noted in the CCRT arm, including leukopenia and emesis, compliance with the combined treatment was good. The second cycle of concurrent chemotherapy was refused by nine patients and was delayed for > or = 1 week for another nine patients. There were no treatment-related deaths in either arm. CONCLUSION: We conclude that CCRT is superior to RT alone for patients with advanced NPC in endemic areas. PMID- 12586798 TI - Vaccination with irradiated autologous tumor cells engineered to secrete granulocyte-macrophage colony-stimulating factor augments antitumor immunity in some patients with metastatic non-small-cell lung carcinoma. AB - PURPOSE: We demonstrated that vaccination with irradiated tumor cells engineered to secrete granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulates potent, specific, and long-lasting antitumor immunity in multiple murine models and patients with metastatic melanoma. To test whether this vaccination strategy enhances antitumor immunity in patients with metastatic non-small-cell lung cancer (NSCLC), we conducted a phase I clinical trial. PATIENTS AND METHODS: Resected metastases were processed to single-cell suspension, infected with a replication-defective adenoviral vector encoding GM-CSF, irradiated, and cryopreserved. Individual vaccines consisted of 1 x 10(6), 4 x 10(6), or 1 x 10(7) cells, depending on overall yield, and were administered intradermally and subcutaneously at weekly and biweekly intervals. RESULTS: Vaccines were successfully manufactured for 34 (97%) of 35 patients. The average GM-CSF secretion was 513 ng/10(6) cells/24 h. Toxicities were restricted to grade 1 to 2 local skin reactions. Nine patients were withdrawn early because of rapid disease progression. Vaccination elicited dendritic cell, macrophage, granulocyte, and lymphocyte infiltrates in 18 of 25 assessable patients. Immunization stimulated the development of delayed-type hypersensitivity reactions to irradiated, dissociated, autologous, nontransfected tumor cells in 18 of 22 patients. Metastatic lesions resected after vaccination showed T lymphocyte and plasma cell infiltrates with tumor necrosis in three of six patients. Two patients surgically rendered as having no evidence of disease at enrollment remain free of disease at 43 and 42 months. Five patients showed stable disease durations of 33, 19, 12, 10, and 3 months. One mixed response was observed. CONCLUSION: Vaccination with irradiated autologous NSCLC cells engineered to secrete GM-CSF enhances antitumor immunity in some patients with metastatic NSCLC. PMID- 12586800 TI - Treatment of localized nonorbital, nonparameningeal head and neck rhabdomyosarcoma: lessons learned from intergroup rhabdomyosarcoma studies III and IV. AB - PURPOSE: The characteristics and clinical outcomes of children and adolescents with localized nonorbital, nonparameningeal head and neck rhabdomyosarcoma (RMS) treated on national protocols from the Intergroup Rhabdomyosarcoma Group are reported. PATIENTS AND METHODS: We conducted a retrospective review of 164 children and adolescents enrolled in the third and fourth Intergroup Rhabdomyosarcoma Studies. Variables analyzed included age, sex, primary tumor site, histologic subtype, clinical group, therapy, site and rate of treatment failure, and time to initial recurrence. RESULTS: Localized nonorbital, nonparameningeal RMS accounted for 9% of all cases of RMS. The median age at diagnosis was 5 years; the median follow-up was 6.6 years. Estimated 5-year failure-free survival (FFS) and survival (S) rates were 76% (95% CI, 69% to 83%) and 83% (95% CI, 77% to 89%), respectively. In multivariate analysis, patients with clinically involved regional nodes (N1) had worse FFS (P =.02). For patients with embryonal tumors, FFS was significantly improved, especially among patients with Group I/II without nodal disease clinical Group I, II N0. For patients with alveolar/undifferentiated histology, FFS was significantly worse in children under the age of 1 year. Actuarial estimates of recurrences at 15 years were local (19%), regional (5%), and distant (9%). CONCLUSION: More than 80% of patients with RMS of the head and neck are cured of their disease using surgery and vincristine, dactinomycin +/- cyclophosphamide with or without radiotherapy. Our results indicate that early, limited exposure to cyclophosphamide might reduce recurrence in low-risk embryonal patients and that reduced dosages might achieve comparable results with improved toxicity profiles. These hypotheses will be tested in the next generation of trials from the Soft Tissue Committee of the Children's Oncology Group. PMID- 12586801 TI - Phase II trial of temozolomide in patients with progressive low-grade glioma. AB - PURPOSE: Temozolomide (Temodar; Schering-Plough Corp, Kenilworth, NJ) is an imidazole tetrazinone that undergoes chemical conversion to the active methylating agent 5-(3-methyltriazen-1yl)imidazole-4-carboximide under physiologic conditions. Previous studies have confirmed activity of Temodar in the treatment of progressive and newly diagnosed malignant gliomas. We have extended these results, and now we report results of a phase II trial of Temodar for patients with progressive, low-grade glioma. PATIENTS AND METHODS: Temodar was administered orally once a day for five consecutive days (in a fasting state) at a starting dose of 200 mg/m(2)/d. Treatment cycles were repeated every 28 days following the first daily dose of Temodar. Response criteria used a combination of magnetic resonance imaging and physical examination to evaluate activity. RESULTS: Forty-six patients with low-grade glioma have been treated to date. The objective response rate was 61% (24% complete response and 37% partial response), with an additional 35% of patients having stable disease. Median progression-free survival (PFS) was 22 months (95% confidence interval [CI], 15 to infinity months) with a 6-month PFS of 98% (95% CI, 94% to 100%) and a 12-month PFS of 76% (95% CI, 63% to 92%). Toxicity observed during the study was limited to only six patients. Three patients experienced grade 3 neutropenia, with a duration greater than 3 weeks in one patient, and two patients experienced grade 3 thrombocytopenia. One patient experienced > or = grade 4 toxicity, with intracerebral hemorrhage, neutropenia, thrombocytopenia, sepsis, and death. CONCLUSION: Initial results indicate that Temodar may be active in the treatment of low-grade glioma, and thus, further evaluation of this agent in the treatment of these tumors is warranted. PMID- 12586802 TI - Phase II study of the efficacy and safety of cisplatin-epinephrine injectable gel administered to patients with unresectable hepatocellular carcinoma. AB - PURPOSE: To study the efficacy and safety of percutaneous cisplatin-epinephrine (CDDP-EPI) injectable gel in patients with localized unresectable hepatocellular carcinoma (HCC). PATIENTS AND METHODS: Eligible patients had histologically proven HCC, no prior treatment except for surgery, and no more than three tumors (each measured < or = 7 cm, total tumor volume < or = 200 cm(3)). They were treated percutaneously under ultrasound or computed tomography (CT) guidance, with up to 10 mL of CDDP-EPI gel (1 mL contains 4 mg of CDDP and 0.1 mg of EPI) per treatment and four treatments in 6 weeks to a maximum of eight treatments. The primary end points were tumor response, defined by change of percentage of tumor necrosis according to CT criteria, and safety. Survival parameters were secondary end points. RESULTS: From June 1997 to April 2000, 58 patients (median age, 65 years) entered the study. All patients were assessable for safety, and 51 were assessable for efficacy. The median number of treatments was four (range, one to eight treatments). Objective response rate was 53% (27 of 51 patients), including 16 complete and 11 partial responses. Of the 27 responders, 14 (52%) subsequently developed progressive disease, but in most of them (93%), a new tumor arose at untreated liver sites. Median survival was 27 months (range, 18.4 to 35.7 months). The 1-, 2-, and 3-year survival rates were 79%, 56%, and 14% respectively. The procedure was well tolerated with only minor side effects. CONCLUSION: Percutaneous local ablation with CDDP-EPI injectable gel can induce significant tumor necrosis and local control for localized unresectable HCC, and the treatment is well tolerated. PMID- 12586803 TI - Dysadherin overexpression in pancreatic ductal adenocarcinoma reflects tumor aggressiveness: relationship to e-cadherin expression. AB - PURPOSE: The E-cadherin-mediated cell adhesion system is frequently inactivated by multiple mechanisms and is involved in tumor progression in many types of cancer. Recently, we reported the cloning and characterization of dysadherin and showed that it downregulated E-cadherin and promoted metastasis. The aim of this study was to investigate the clinical significance of dysadherin expression and the relationship between dysadherin expression and E-cadherin expression in pancreatic ductal adenocarcinoma. PATIENTS AND METHODS: We examined dysadherin and E-cadherin expression in 125 surgically resected pancreatic ductal adenocarcinoma patients using immunohistochemistry. RESULTS: Dysadherin was expressed at the cell membrane of cancer cells, but not in nontumor duct and acinar cells. Its expression was stronger in infiltrative and poorly differentiated nests compared with well-differentiated nests. Although the correlation between the expression of dysadherin and E-cadherin was not significant, a group of patients showed reduced E-cadherin expression with dysadherin overexpression. Increased dysadherin expression was significantly correlated with distant metastasis (P =.047), high tumor grade (P =.006), positive tumor margins (P =.024), and infiltrative type of growth pattern (P =.014). A survival advantage was observed in patients with 0% to 20% dysadherin positive cells compared with patients with 51% to 100% dysadherin-positive cells, independent of tumor-node-metastasis classification, and World Health Organization tumor grade (P =.019). A combination of increased dysadherin expression and reduced E-cadherin expression (< 90%) further worsened the prognosis. CONCLUSION: In pancreatic ductal adenocarcinoma, dysadherin expression seems to reflect tumor aggressiveness and to be a positive marker of poor prognosis when considered both alone and in combination with downregulation of E cadherin. PMID- 12586804 TI - Effect of lymphatic mapping on the new tumor-node-metastasis classification for colorectal cancer. AB - PURPOSE: Sensitive detection methods and accurate reporting are necessary to determine the prognostic significance of micrometastases (MM) and isolated tumor cells (ITCs) in lymph nodes that drain colorectal cancers (CRCs). This study examined the role of lymphatic mapping (LM) in the application of the new tumor node-metastasis (TNM) classification for MM and ITC. PATIENTS AND METHODS: All patients at the John Wayne Cancer Institute underwent LM immediately before standard resection of primary CRC between 1996 and 2001. Sentinel nodes (SNs) were identified using blue dye and/or radiotracer and were examined by hematoxylin-eosin (H&E) staining, cytokeratin immunohistochemistry, and multilevel sectioning. The comparison group comprised 370 patients whose primary CRCs were resected without LM during the same period at the same institution. RESULTS: LM was successfully performed in 115 of 120 (96%) patients and correctly predicted the tumor status of the nodal basin in 110 of 115 (96%) patients. Thirty-seven patients (32%) were lymph node-positive by H&E, ITC and MM were found in 23 patients (29.4%) whose lymph nodes were negative by H&E. Tumor deposits were found in the SN only in 29 patients (50%). Nodal involvement was identified for 14.3%, 30%, 74.6%, and 83.3% of T1, T2, T3, and T4 tumors, respectively, in the study group, and for 6.8%, 8.5%, 49.3%, and 41.8% of T1, T2, T3, and T4 tumors, respectively, in the comparison group. The study group had a higher percentage of nodal metastases (53% v 36%; P <.01) and a higher incidence of MM and ITC (29.4% v 1.9%; P <.0001). The mean number of lymph nodes found in the study group (14) was also significantly more than the number found in the comparison group (10; P <.00001). CONCLUSION: Conventional examination of lymph nodes for CRC is inadequate for the detection of MM and ITC as described in the new TNM classification. Thus, LM and focused SN analysis should be considered to fully stage CRC. PMID- 12586805 TI - Health-related quality-of-life parameters as independent prognostic factors in advanced or metastatic bladder cancer. AB - PURPOSE: This retrospective analysis examined prognostic significance of health related quality-of-life (HRQoL) parameters combined with baseline clinical factors on outcomes (overall survival, time to progressive disease, and time to treatment failure) in bladder cancer. PATIENTS AND METHODS: Outcome and HRQoL (European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C30) data were collected prospectively in a phase III study assessing gemcitabine and cisplatin versus methotrexate, vinblastine, doxorubicin, and cisplatin in locally advanced or metastatic bladder cancer. Prespecified baseline clinical factors (performance status, tumor-node-metastasis staging, visceral metastases [VM], alkaline phosphatase [AP] level, number of metastatic sites, prior radiotherapy, disease measurability, sex, time from diagnosis, and sites of disease) and selected HRQoL parameters (global QoL; all functional scales; symptoms: pain, fatigue, insomnia, dyspnea, anorexia) were evaluated using Cox's proportional hazards model. Factors with individual prognostic value (P <.05) on outcomes in univariate models were assessed for joint prognostic value in a multivariate model. A final model was developed using a backward selection strategy. RESULTS: Patients with baseline HRQoL were included (364 of 405, 90%). The final model predicted longer survival with low/normal AP levels, no VM, high physical functioning, low role functioning, and no anorexia. Positive prognostic factors for time to progressive disease were good performance status, low/normal AP levels, no VM, and minimal fatigue; for time to treatment failure, they were low/normal AP levels, minimal fatigue, and no anorexia. Global QoL was a significant predictor of outcome in univariate analyses but was not retained in the multivariate model. CONCLUSION: HRQoL parameters are independent prognostic factors for outcome in advanced bladder cancer; their prognostic importance needs further evaluation. PMID- 12586806 TI - Effect of endothelin-A receptor blockade with atrasentan on tumor progression in men with hormone-refractory prostate cancer: a randomized, phase II, placebo controlled trial. AB - PURPOSE: To evaluate the efficacy and safety of atrasentan (ABT-627), an endothelin-A receptor antagonist, in the treatment of asymptomatic, hormone refractory prostatic adenocarcinoma. PATIENTS AND METHODS: A double-blind, randomized, placebo-controlled clinical trial of hormone-refractory prostate cancer (HRPCa) patients was conducted in the United States and Europe. Two hundred eighty-eight asymptomatic patients with HRPCa and evidence of metastatic disease were randomly assigned to one of three study groups receiving a once daily oral dose of placebo, 2.5 mg atrasentan, or 10 mg atrasentan, respectively. Primary end point was time to progression; secondary end points included time to prostate-specific antigen (PSA) progression, bone scan changes, and changes in bone and tumor markers. RESULTS: The three treatment groups were similar in all baseline characteristics. Median time to progression in intent-to-treat (ITT) patients (n = 288) was longer in the 10-mg atrasentan group compared with the placebo group: 183 v 137 days, respectively; (P =.13). Median time to progression in evaluable patients (n = 244) was significantly prolonged, from 129 days (placebo group) to 196 days (10-mg atrasentan group; P =.021). For both ITT and evaluable populations in the 10-mg atrasentan group, median time to PSA progression was twice that of the placebo group (155 v 71 days; P =.002). Patients who received placebo continued to have significant increases from baseline in serum (lactate dehydrogenase [LDH]), a marker of disease burden; elevations in LDH were uniformly attenuated by atrasentan in the ITT population. Headache, peripheral edema, and rhinitis were primary side effects, typically of mild to moderate severity. Quality of life was not adversely affected by atrasentan. CONCLUSION: Atrasentan is an oral, targeted therapy with favorable tolerability and the potential to delay progression of HRPCa. PMID- 12586807 TI - Radical cystectomy for bladder cancer today--a homogeneous series without neoadjuvant therapy. AB - PURPOSE: To investigate the effect of pelvic lymph node dissection and radical cystectomy for transitional cell cancer of the bladder on recurrence-free and overall survival, pelvic recurrences, and metastatic patterns in a homogeneous group. PATIENTS AND METHODS: A consecutive series of patients undergoing pelvic lymphadenectomy and radical cystectomy between 1985 and 2000 was analyzed. All patients were staged N0, M0 preoperatively, and no patient received neoadjuvant radio/chemotherapy. Pathologic characteristics based on the 1997 tumor-node metastasis system, recurrence-free/overall survival, and metastatic patterns were determined. RESULTS: Five hundred seven patients (age 66 +/- 12 years) with a mean follow-up time of 45 months (range, 0.1 to 176 months) were analyzed. Five year recurrence-free and overall survival were, respectively, 73% and 62% for patients with organ-confined, lymph node-negative tumors (n = 217; < or = pT2, pN0) and 56% and 49% for non-organ-confined, lymph node-negative tumors (n = 166; > pT2, pN0). Positive lymph nodes were found in 124 (24%) patients who had a 5 year recurrence-free (33%) or overall (26%) survival. Isolated local recurrences were observed in 3% of patients with organ-confined tumors (< or = pT2, pN0), 11% with non-organ-confined tumors (> pT2, pN0), and 13% with positive lymph nodes (any pT, pN+). Distant metastases developed in 25% of patients with organ confined tumors, 37% with non-organ-confined tumors, and 51% with positive lymph nodes. CONCLUSION: Despite negative preoperative staging, pelvic lymphadenectomy and cystectomy for bladder cancer reveal a high percentage of unsuspected nodal metastases (24%) that have a 25% chance for long-term survival. This procedure also ensures a low pelvic recurrence rate even in lymph node-positive patients, and patients with locally advanced cancer have a 56% probability of 5-year recurrence-free survival. PMID- 12586808 TI - Intravesical gemcitabine therapy for superficial transitional cell carcinoma of the bladder: a phase I and pharmacokinetic study. AB - PURPOSE: To determine maximum-tolerated dose, toxicities, and pharmacokinetics associated with weekly intravesical gemcitabine therapy in patients with superficial bladder cancer. PATIENTS AND METHODS: Fifteen patients with recurrent superficial transitional cell bladder carcinoma who experienced prior intravesical therapy failure were studied. Two to 4 weeks after complete transurethral resection, gemcitabine was administered intravesically, once weekly for 6 consecutive weeks. Dwell time was 2 hours. Pharmacokinetics of gemcitabine and its metabolite, 2'2'-difluorodeoxyuridine (dFdU), were studied in plasma and urine. Cystoscopy was repeated 6 weeks after therapy. RESULTS: Three-patient cohorts were enrolled sequentially at doses of 500, 1,000, and 1,500 mg in 100 mL 0.9% NaCl. Two patients received 2,000 mg in 100 mL. An additional four patients received 2,000 mg in 50 mL. No grade 4 toxicity or clinically relevant myelosuppression was noted. Nine of 13 evaluable patients were recurrence-free at 12 weeks. Low concentrations of gemcitabine (< or = 1 microg/mL) were present transiently in plasma of all patients receiving 2,000 mg in 50 mL. Gemcitabine was undetectable in plasma of other patients. dFdU was undetectable in plasma of patients receiving less than 1,500 mg. At doses > or = 1,500 mg, dFdU concentrations increased until 90 to 120 minutes and then declined little, if any. Plasma dFdU concentrations implied absorption of 0.5% to 5.5% of instilled dose. Between 61% and 100% of the gemcitabine dose was accounted for in voided urine. No dFdU was measured in voided urine. CONCLUSION: Intravesical gemcitabine, at doses up to 2 g/wk, is well tolerated, is associated with minimal systemic absorption, and has promising efficacy in treatment of superficial bladder cancer. PMID- 12586809 TI - Importance of minimal residual disease testing during the second year of therapy for children with acute lymphoblastic leukemia. AB - PURPOSE: A high level of minimal residual disease (MRD) after induction chemotherapy in children with acute lymphoblastic leukemia (ALL) is an indicator of relative chemotherapy resistance and a risk factor for relapse. However, the significance of MRD in the second year of therapy is unclear. Moreover, it is unknown whether treatment intervention can alter outcome in patients with detectable MRD. PATIENTS AND METHODS: We assessed the prognostic value of MRD testing in bone marrow samples from 85 children at 1, 12, and 24 months from diagnosis using clone-specific polymerase chain reaction primers designed to detect clonal antigen receptor gene rearrangements. These children were part of a multicenter, randomized clinical trial, which, in the second year of treatment, compared a 2-month reinduction-reintensification followed by maintenance chemotherapy with standard maintenance chemotherapy alone. RESULTS: MRD was detected in 69% of patients at 1 month, 25% at 12 months, and 28% at 24 months from diagnosis. By univariate analysis, high levels of MRD at 1 month, or the presence of any detectable MRD at 12 or 24 months from diagnosis, were highly predictive of relapse. Multivariate analysis showed that MRD testing at 1 and 24 months each had independent prognostic significance. Intensified therapy at 12 months from diagnosis did not improve prognosis in those patients who were MRD positive at 12 months from diagnosis. CONCLUSION: Clinical outcome in childhood ALL can be predicted with high accuracy by combining the results of MRD testing at 1 and 24 months from diagnosis. PMID- 12586810 TI - Postrelapse survival in osteosarcoma of the extremities: prognostic factors for long-term survival. AB - PURPOSE: To identify factors that influence postrelapse survival (PRS) in patients with nonmetastatic osteosarcoma of the extremity. PATIENTS AND METHODS: One hundred sixty-two patients with recurrent osteosarcoma of the extremity were retrospectively reviewed. The first-line treatment included surgery of the primary lesion and chemotherapy with methotrexate, doxorubicin, cisplatin, and ifosfamide. RESULTS: The projected 5-year PRS rate was 28%. Patients who had complete surgery of recurrence had a 5-year PRS of 39%, whereas for those who did not have complete surgery, PRS was 0% at 3 years (P <.0001). In the latter group, PRS was not influenced by site of recurrence and relapse-free interval (RFI), although it was influenced (P =.006) by the use of second-line chemotherapy (PRS, 53% at 12 months for patients who received chemotherapy v 12% for those who did not). In patients who had complete surgery, PRS was influenced by site of relapse (5-year PRS, lung 44%, other 19%; P <.06), RFI (5-year PRS at < or = 24 months, 20%; at > 24 months, 60%; P <.0001), and number of lung metastases (5-year PRS, two or fewer nodules, 59%; more than two nodules, 14%; P <.0001) but not by the use of a second-line chemotherapy treatment. CONCLUSION: RFI, site of metastases, and number of pulmonary nodules are the main prognostic factors for PRS in osteosarcoma. Complete surgery of recurrence is pivotal in the strategy of treatment. Patients with unresectable recurrence benefit from second-line chemotherapy, whereas our data do not support a generalized use of chemotherapy after complete surgery of first recurrence. PMID- 12586811 TI - Pregnancy outcome of partners of male survivors of childhood cancer: a report from the Childhood Cancer Survivor Study. AB - PURPOSE: This study was undertaken to determine the effect, if any, on pregnancy loss, live births, and birthweight of treatment for cancer diagnosed during childhood or adolescence. PATIENTS AND METHODS: We reviewed pregnancy outcome among sexually active male Childhood Cancer Survivor Study (CCSS) participants who responded to a questionnaire before February 3, 2000. Medical records of all members of the cohort were abstracted to obtain chemotherapeutic agents administered, the cumulative dose of drug administered for several drugs of interest, and the doses, volumes, and dates of administration of all radiotherapy. RESULTS: There were 4,106 sexually active males; 1,227 reported they sired 2,323 pregnancies (69% live births, 1% stillbirths, 13% miscarriages, 13% abortions, 5% unknown or in gestation). The male-to-female ratio of the offspring of the partners of the male survivors was significantly different from that of the offspring of the partners of the male siblings of the survivors (1.0:1.03 v 1.24:1.0) (P =.016). The proportion of pregnancies of the partners of male survivors that ended with a liveborn infant was significantly lower than for the partners of the male siblings of the survivors who were the control group for comparison (relative risk = 0.77, P =.007). There were no significant differences in pregnancy outcome by treatment. CONCLUSION: This large study did not identify adverse pregnancy outcomes for the partners of male survivors treated with most chemotherapeutic agents. The reversal of the sex ratio and the association observed for procarbazine warrant further investigation. PMID- 12586812 TI - Skp2 protein expression in soft tissue sarcomas. AB - BACKGROUND: p45 S phase kinase-associated protein-2 (p45(skp2)), a member of the F-box family of proteins, is an important component of the Skp1-Cullin-F-box protein (SCF) ubiquitin-ligase complex (SCF(skp2)). The latter has been implicated in the ubiquitination and degradation of p27(kip1) (p27) and G(1)-S cell cycle progression. The expression and prognostic role of Skp2 in a large series of soft tissue sarcomas has not been previously investigated. METHODS: Clinicopathologic features and immunohistochemical expression of Skp2, p27, and Ki-67 proteins were studied in 182 cases of soft tissue sarcomas (American Joint Committee on Cancer stages II and III). Survival analyses were performed using the Kaplan-Meier method and the Cox regression model. RESULTS: The male to female ratio was 1.2:1, and the median age at the diagnosis was 53 years. The tumors were predominantly located in the lower extremities (n = 163; 90%) and had a median size of 9 cm. High Skp2 expression (> or = 10% of the cells) was identified in 68 tumors (37%), and was correlated with high grade histology (P =.002) and Ki-67 proliferative index (r = 0.44; P <.0001), but not with p27 expression (r = -0.02; P =.80). By univariate analysis, high Skp2 expression was associated with decreased metastasis-free, disease-free, and overall survival. In a multivariate model, high Skp2 expression was an independent predictor for decreased local recurrence-free, disease-free, and overall survival. CONCLUSION: These results indicate that Skp2 expression is associated with cell proliferation and a worse prognosis in soft tissue sarcomas. The lack of an inverse correlation between Skp2 and p27 suggests that additional molecular events associated with either Skp2 expression or p27 proteolysis may be operating in these tumors. PMID- 12586813 TI - Barriers to hospice care among older patients dying with lung and colorectal cancer. AB - PURPOSE: To identify factors associated with hospice enrollment and length of stay in hospice among patients dying with lung or colorectal cancer. METHODS: We used the Linked Medicare-Tumor Registry Database to conduct a retrospective analysis of the last year of life among Medicare beneficiaries diagnosed with lung or colorectal cancer at age > or = 66 years between January 1, 1973, and December 31, 1996, in the Surveillance, Epidemiology, and End Results Program who died between January 1, 1988, and December 31, 1998. Our outcomes of interest were time from cancer diagnosis to hospice enrollment and length of stay in hospice care. We used Cox proportional hazards regression to adjust for demographic and clinical information. RESULTS: We studied elderly patients dying with lung cancer (n = 62,117) or colorectal cancer (n = 57,260). Overall, 27% of patients (n = 16,750) with lung cancer and 20% of patients (n = 11,332) with colorectal cancer received hospice care before death. Median length of stay for hospice patients with lung and colorectal cancer was 25 and 28 days, respectively. Overall, 20% of patients entered hospice within 1 week of death, whereas 6% entered more than 6 months before death. Factors associated with later hospice enrollment include being male; being of nonwhite, nonblack race; having fee-for-service insurance; and residing in a rural community. Many of these factors also were associated with shorter stays in hospice. CONCLUSION: Although use of hospice care has increased dramatically over time, specific patient groups, including men, patients residing in rural communities, and patients with fee-for-service insurance continue to experience delays in hospice enrollment. PMID- 12586814 TI - Minidose warfarin prophylaxis for catheter-associated thrombosis in cancer patients: can it be safely associated with fluorouracil-based chemotherapy? AB - PURPOSE: The use of prophylactic low-dose oral warfarin in cancer patients with a central venous catheter (CVC) in place has an established role in the prevention of thrombotic complications and is associated with a low hemorrhagic risk. Despite the literature indicating an adverse interaction between warfarin and fluorouracil (FU), the frequency of this interaction and whether it occurs when minidose warfarin is used is unknown. We analyzed the incidence of alterations in the International Normalized Ratio (INR) and bleeding in cancer patients given minidose warfarin during treatment with continuous-infusion FU-based regimens. PATIENTS AND METHODS: Between July 1999 and August 2001, 95 cancer patients were evaluated. Forty-one patients (43%) had liver metastases. Seventy-nine patients (83%) had a Groshong CVC (Bard Access System, Salt Lake City, UT), and 16 (17%) had a Port-a-Cath device (Bard Access System). All patients received oral warfarin at a dose of 1 mg/daily as prophylaxis beginning the day after the catheter was positioned. An INR of more than 1.5 was considered significantly elevated. RESULTS: INR elevation occurred in 31 patients (33%), with 18 patients (19%) having an INR more than 3.0. Twelve (39%) of the 31 patients had liver metastases. Bleeding was observed in eight patients (8%); seven of these patients had elevated INR levels. We observed INR elevations in 12 of 21 patients treated with a FU, folinic acid, and oxaliplatin (FOLFOX) regimen, 11 of 40 treated with a de Gramont regimen (FU and folinic acid), and five of 19 treated with a FU, folinic acid, and irinotecan (FOLFIRI) regimen. CONCLUSION: A high incidence of INR abnormalities was observed in our cohort of patients, especially those treated with FOLFOX regimen. Clinicians should be aware of this interaction and should regularly monitor the prothrombin time in patients receiving warfarin and FU. PMID- 12586816 TI - Comprehensive geriatric evaluation in elderly patients with lymphoma: feasibility of a patient-tailored treatment plan. PMID- 12586815 TI - Hereditary breast-ovarian cancer at the bedside: role of the medical oncologist. AB - PURPOSE: To provide practical considerations for diagnosing, counseling, and managing patients at high risk for hereditary breast cancer. DESIGN: We have studied 98 extended hereditary breast cancer (HBC)/hereditary breast-ovarian cancer (HBOC) families with BRCA1/2 germline mutations. From these families, 1,315 individuals were counseled and sampled for DNA testing. Herein, 716 of these individuals received their DNA test results in concert with genetic counseling. Several challenging pedigrees were selected from Creighton University's hereditary cancer family registry, as well as one family from Evanston/Northwestern Healthcare, to be discussed in this present report. RESULTS: Many obstacles were identified in diagnosis, counseling, and managing patients at high risk for HBC/HBOC. These obstacles were early noncancer death of key relatives, perception of insurance or employment discrimination, fear, anxiety, apprehension, reduced gene penetrance, and poor compliance. Other important issues such as physician culpability and malpractice implications for failure to collect or act on the cancer family history were identified. CONCLUSION: When clinical gene testing emerged for BRCA1 and BRCA2, little was known about the efficacy of medical interventions. Potential barriers to uptake of testing were largely unexplored. Identification and referral of high-risk patients and families to genetic counseling can greatly enhance the care of the population at the highest risk for cancer. However, because premonitory physical stigmata are absent in most of these syndromes, an HBOC diagnosis may be missed unless a careful family history of cancer of the breast, ovary, or several integrally associated cancers is obtained. PMID- 12586818 TI - Timing is everything, or could be. PMID- 12586820 TI - Two subtypes of ecdysis-triggering hormone receptor in Drosophila melanogaster. AB - Insect ecdysis is a hormonally programmed physiological sequence that enables insects to escape their old cuticle at the end of each developmental stage. The immediate events leading to ecdysis, which are initiated upon release of ecdysis triggering hormones (ETH) into the bloodstream, include respiratory inflation and sequential stereotypic behaviors that facilitate shedding of the cuticle. Here we report that the Drosophila gene CG5911 encodes two functionally distinct subtypes of G protein-coupled receptors through alternative splicing (CG5911a and CG5911b) that respond preferentially to ecdysis-triggering hormones of flies and moths. These subtypes show differences in ligand sensitivity and specificity, suggesting that they may play separate roles in ETH signaling. At significantly higher concentrations (>100-fold), certain insect and vertebrate peptides also activate these receptors, providing evidence that CG5911 is evolutionarily related to the thyrotropin-releasing hormone and neuromedin U receptors. The ETH signaling system in insects is a vital system that provides opportunities for the construction of models for the molecular basis of stereotypic animal behavior as well as a target for the design of more sophisticated insect-selective pest control strategies. PMID- 12586821 TI - Reduced affinity for Isoniazid in the S315T mutant of Mycobacterium tuberculosis KatG is a key factor in antibiotic resistance. AB - Catalase-peroxidase (KatG) from Mycobacterium tuberculosis is responsible for the activation of the antitubercular drug isonicotinic acid hydrazide (INH) and is important for survival of M. tuberculosis in macrophages. Characterization of the structure and catalytic mechanism of KatG is being pursued to provide insights into drug (INH) resistance in M. tuberculosis. Site-directed mutagenesis was used to prepare the INH-resistant mutant KatG[S315T], and the overexpressed enzyme was characterized and compared with wild-type KatG. KatG[S315T] exhibits a reduced tendency to form six-coordinate heme, because of coordination of water to iron during purification and storage, and also forms a highly unstable Compound III (oxyferrous enzyme). Catalase activity and peroxidase activity measured using t butylhydroperoxide and o-dianisidine were moderately reduced in the mutant compared with wild-type KatG. Stopped-flow spectrophotometric experiments revealed a rate of Compound I formation similar to wild-type KatG using peroxyacetic acid to initiate the catalytic cycle, but no Compound I was detected when bulkier peroxides (chloroperoxybenzoic acid, t-butylhydroperoxide) were used. The affinity of resting (ferric) KatG[S315T] for INH, measured using isothermal titration calorimetry, was greatly reduced compared with wild-type KatG, as were rates of reaction of Compound I with the drug. These observations reveal that although KatG[S315T] maintains reasonably good steady state catalytic rates, poor binding of the drug to the enzyme limits drug activation and brings about INH resistance. PMID- 12586822 TI - Tamalin is a scaffold protein that interacts with multiple neuronal proteins in distinct modes of protein-protein association. AB - Tamalin is a scaffold protein that comprises multiple protein-interacting domains, including a 95-kDa postsynaptic density protein (PSD-95)/discs-large/ZO 1 (PDZ) domain, a leucine-zipper region, and a carboxyl-terminal PDZ binding motif. Tamalin forms a complex with metabotropic glutamate receptors and guanine nucleotide exchange factor cytohesins and promotes intracellular trafficking and cell surface expression of group 1 metabotropic glutamate receptors. In the present study, using several different approaches we have shown that tamalin interacts with multiple neuronal proteins through its distinct protein-binding domains. The PDZ domain of tamalin binds to the PDZ binding motifs of SAP90/PSD 95-associated protein and tamalin itself, whereas the PDZ binding motif of tamalin is capable of interacting with the PDZ domain of S-SCAM. In addition, tamalin forms a complex with PSD-95 and Mint2/X11beta/X11L by mechanisms different from the PDZ-mediated interaction. Tamalin has the ability to assemble with these proteins in vivo; their protein complex with tamalin was verified by coimmunoprecipitation of rat brain lysates. Interestingly, the distinct protein interacting domains of tamalin are evolutionarily conserved, and mRNA expression is developmentally up-regulated at the postnatal period. The results indicate that tamalin exists as a key element that forms a protein complex with multiple postsynaptic and protein-trafficking scaffold proteins. PMID- 12586823 TI - Mitochondrial Ca2+ uptake requires sustained Ca2+ release from the endoplasmic reticulum. AB - We analyzed the role of inositol 1,4,5-trisphosphate-induced Ca(2+) release from the endoplasmic reticulum (ER) (i) in powering mitochondrial Ca(2+) uptake and (ii) in maintaining a sustained elevation of cytosolic Ca(2+) concentration ([Ca(2+)](c)). For this purpose, we expressed in HeLa cells aequorin-based Ca(2+) sensitive probes targeted to different intracellular compartments and studied the effect of two agonists: histamine, acting on endogenous H(1) receptors, and glutamate, acting on co-transfected metabotropic glutamate receptor (mGluR1a), which rapidly inactivates through protein kinase C-dependent phosphorylation and thus causes transient inositol 1,4,5-trisphosphate production. Glutamate induced a transient [Ca(2+)](c) rise and drop in ER luminal [Ca(2+)] ([Ca(2+)](er)), and then the ER refilled with [Ca(2+)](c) at resting values. With histamine, [Ca(2+)](c) after the initial peak stabilized at a sustained plateau, and [Ca(2+)](er) decreased to a low steady-state value. In mitochondria, histamine evoked a much larger mitochondrial Ca(2+) response than glutamate ( approximately 15 versus approximately 65 microm). Protein kinase C inhibition, partly relieving mGluR1a desensitization, reestablished both the [Ca(2+)](c) plateau and the sustained ER Ca(2+) release and markedly increased the mitochondrial Ca(2+) response. Conversely, mitochondrial Ca(2+) uptake evoked by histamine was drastically reduced by very transient ( approximately 2-s) agonist applications. These data indicate that efficient mitochondrial Ca(2+) uptake depends on the preservation of high Ca(2+) microdomains at the mouth of ER Ca(2+) release sites close to mitochondria. This in turn depends on continuous Ca(2+) release balanced by Ca(2+) reuptake into the ER and maintained by Ca(2+) influx from the extracellular space. PMID- 12586824 TI - A broken alpha -helix in folded alpha -Synuclein. AB - alpha-Synuclein is a small cytosolic protein of presynaptic nerve terminals composed of seven 11-residue repeats and a hydrophilic tail. alpha-Synuclein misfolding and dysfunction may contribute to the pathogenesis of Parkinson's disease and neurodegenerative dementias, but its normal folding and function are unknown. In solution, alpha-synuclein is natively unstructured but assumes an alpha-helical conformation upon binding to phospholipid membranes. We now show that this conformation of alpha-synuclein consists of two alpha-helical regions that are interrupted by a short break. The structural organization of the alpha helices of alpha-synuclein was not anticipated by sequence analyses and may be important for its pathogenic role. PMID- 12586825 TI - An Ile-568 to Asn polymorphism prevents normal trafficking and function of the human P2X7 receptor. AB - The P2X(7) receptor is a ligand-gated channel that is highly expressed on mononuclear cells and that mediates ATP-induced apoptosis of these cells. Wide variations in the function of the P2X(7) receptor have been observed, in part because of a loss-of-function polymorphism that changes Glu-496 to Ala without affecting the surface expression of the receptor on lymphocytes. In this study a second polymorphism (Ile-568 to Asn) has been found in heterozygous dosage in three of 85 normal subjects and in three of 45 patients with chronic lymphocytic leukemia. P2X(7) function was measured by ATP-induced fluxes of Rb(+), Ba(2+), and ethidium(+) into various lymphocyte subsets and was decreased to values of approximately 25% of normal. The expression of the P2X(7) receptor on lymphocytes was approximately half that of normal values as measured by the binding of fluorescein-conjugated monoclonal antibody. Transfection experiments showed that P2X(7) carrying the Ile-568 to Asn mutation was non-functional because of the failure of cell surface expression. The differentiation of monocytes to macrophages with interferon-gamma up-regulated P2X(7) function in cells heterozygous for the Ile-568 to Asn mutation to a value around 50% of normal. These data identify a second loss-of-function polymorphism within the P2X(7) receptor and show that Ile-568 is critical to the trafficking domain, which we have shown to lie between residues 551 and 581. PMID- 12586826 TI - Specific expression and regulation of the new melanoma inhibitory activity related gene MIA2 in hepatocytes. AB - The novel human gene MIA2 encoding a melanoma inhibitory activity (MIA) homologous protein was identified by a GenBank(TM) search. MIA2, together with MIA, OTOR, and TANGO, belongs to the novel MIA gene family sharing important structural features, significant homology at both the nucleotide and protein levels, and similar genomic organization. In situ hybridization, reverse transcriptase-PCR, and Northern blots presented a highly tissue-specific MIA2 expression pattern in the liver. Promoter studies analyzing transcriptional regulation of MIA2 revealed an HNF-1-binding site at position -236 controlling hepatocyte-specific expression. Mutation of the site led to a complete loss of promoter activity in HepG2 cell. Further sites detected in the MIA2 promoter were consensus binding sites for SMAD and STAT3, Consistently, stimulation of MIA2 mRNA expression occurred by treatment with interleukin-6, transforming growth factor-beta, and conditioned medium from activated hepatic stellate cells. In accordance with these results, MIA2 mRNA was found to be increased in liver tissue of patients with chronic hepatitis C infection compared with controls. MIA2 mRNA levels were significantly higher in patients with severe fibrosis or inflammation than in patients with less severe fibrosis or inflammation. In summary our data indicate that MIA2 represents a potential novel acute phase protein and MIA2 expression responds to liver damage. The increased transcription in more severe chronic liver disease suggests that MIA2 may serve as a marker of hepatic disease activity and severity. PMID- 12586827 TI - A novel EID-1 family member, EID-2, associates with histone deacetylases and inhibits muscle differentiation. AB - An EID-1 (E1A-like inhibitor of differentiation-1) inhibits differentiation by blocking the histone acetyltransferase activity of p300. Here we report a novel inhibitor of differentiation exhibiting homology to EID-1, termed EID-2 (EID-1 like inhibitor of differentiation-2). EID-2 inhibited MyoD-dependent transcription and muscle differentiation. Unlike EID-1, EID-2 did not block p300 activity. Interestingly, EID-2 associated with class I histone deacetylases (HDACs). The N-terminal portion of EID-2 was required for the binding to HDACs. This region was also involved in the transcriptional repression and nuclear localization, suggesting the importance of the involvement of HDACs in the EID-2 function. These results indicate a new family of differentiation inhibitors, although there are several differences in the biochemical mechanisms between EID 2 and EID-1. PMID- 12586828 TI - Role of histone methyltransferase G9a in CpG methylation of the Prader-Willi syndrome imprinting center. AB - Imprinted genes in mammals are often located in clusters whose imprinting is subject to long range regulation by cis-acting sequences known as imprinting centers (ICs). The mechanisms by which these ICs exert their effects is unknown. The Prader-Willi syndrome IC (PWS-IC) on human chromosome 15 and mouse chromosome 7 regulates imprinted gene expression bidirectionally within an approximately 2 megabase region and shows CpG methylation and histone H3 Lys-9 methylation in somatic cells specific for the maternal chromosome. Here we show that histone H3 Lys-9 methylation of the PWS-IC is reduced in mouse embryonic stem (ES) cells lacking the G9a histone H3 Lys-9/Lys-27 methyltransferase and that maintenance of CpG methylation of the PWS-IC in mouse ES cells requires the function of G9a. We show by RNA fluorescence in situ hybridization (FISH) that expression of Snrpn, an imprinted gene regulated by the PWS-IC, is biallelic in G9a -/- ES cells, indicating loss of imprinting. By contrast, Dnmt1 -/- ES cells lack CpG methylation of the PWS-IC but have normal levels of H3 Lys-9 methylation of the PWS-IC and show normal monoallelic Snrpn expression. Our results demonstrate a role for histone methylation in the maintenance of parent-specific CpG methylation of imprinting regulatory regions and suggest a possible role of histone methylation in establishment of these CpG methylation patterns. PMID- 12586829 TI - Cyclosporin A prevents the hypoxic adaptation by activating hypoxia-inducible factor-1alpha Pro-564 hydroxylation. AB - The mechanism by which hypoxia induces gene transcription involves the inhibition of hypoxia-inducible factor (HIF)-1alpha prolyl hydroxylase activity, which prevents von Hippel-Lindau (vHL)-dependent targeting of HIF-1alpha to the ubiquitin-proteasome pathway. HIF-1alpha is stabilized, translocates to the nucleus, interacts with hypoxia-responsive elements, and promotes the activation of target genes. This report shows that cyclosporin A (CsA) interferes with the hypoxic signaling cascade in C6 glioma cells. CsA inhibits hypoxia-dependent gene transcription in a reporter gene assay and prevents the hypoxic accumulation of HIF-1alpha. Addition of the 530-603 C-terminal oxygen-dependent degradation (ODD) domain of HIF-1alpha to the green fluorescent protein (GFP) destabilized the protein in an oxygen-dependent manner. CsA prevented the hypoxic stabilization of an ODD.GFP fusion protein. An assay for 2-oxoglutarate-dependent dioxygenases was developed using a light mitochondrial kidney fraction as a source of enzyme. It uses the capacity of specific peptides to stimulate the degradation of [(14)C]2 oxoglutarate. CsA stimulated the enzymatic activity in the presence of a peptide that mimicked the 557-576 sequence of HIF-1alpha. The enzyme promoted [(35)S]vHL binding to glutathione S-transferase (GST).ODD fusion protein. This association increased in the presence of CsA. CsA effects were not observed when the proline residue corresponding to Pro-564 in the HIF-1alpha sequence was replaced by a hydroxyproline or an alanine residue. Finally, CsA increased vHL-ODD interaction during hypoxia. We conclude that CsA destabilizes HIF-1alpha by promoting hydroxylation of Pro-564 in the ODD domain. Such a mechanism may prevent hypoxic adaptation during CsA-induced nephrotoxicity and contribute to the adverse effects of this drug. PMID- 12586830 TI - R-Ras alters Ca2+ homeostasis by increasing the Ca2+ leak across the endoplasmic reticular membrane. AB - Evidence in the literature implicating both Ras-like Ras (R-Ras) and intracellular Ca(2+) in programmed cell death and integrin-mediated adhesion prompted us to investigate the possibility that R-Ras alters cellular Ca(2+) handling. Chinese hamster ovary cells expressing the cholecystokinin (CCK)-A receptor were loaded with indo-1 to study the effects of constitutively active V38R-Ras and dominant negative N43R-Ras on the kinetics of the thapsigargin (Tg)- and CCK(8)-induced Ca(2+) rises using high speed confocal microscopy. In the absence of extracellular Ca(2+), both 1 microm Tg, a potent and selective inhibitor of the Ca(2+) pump of the intracellular Ca(2+) store, and 100 nm CCK(8) evoked a transient rise in Ca(2+), the size of which was decreased significantly after expression of V38R-Ras. At 0.1 nm, CCK(8) evoked periodic Ca(2+) rises. The frequency of these Ca(2+) oscillations was reduced significantly in V38R-Ras expressing cells. In contrast to V38R-Ras, N43R-Ras did not alter the kinetics of the Tg- and CCK(8)-induced Ca(2+) rises. The present findings are compatible with the idea that V38R-Ras expression increases the passive leak of Ca(2+) of the store leading to a decrease in Ca(2+) content of this store, which, in turn, leads to a decrease in frequency of the CCK(8)-induced cytosolic Ca(2+) oscillations. The effect of V38R-Ras on the Ca(2+) content of the intracellular Ca(2+) store closely resembles that of the antiapoptotic protein Bcl-2 observed earlier. Together with reports on the role of dynamic Ca(2+) changes in integrin mediated adhesion, this leads us to propose that the reduction in endoplasmic reticulum Ca(2+) content may underlie the antiapoptotic effect of R-Ras, whereas the decrease in frequency of stimulus-induced Ca(2+) oscillations may play a role in the inhibitory effect of R-Ras on stimulus-induced cell detachment and migration. PMID- 12586831 TI - Maurocalcine and peptide A stabilize distinct subconductance states of ryanodine receptor type 1, revealing a proportional gating mechanism. AB - Maurocalcine (MCa) isolated from Scorpio maurus palmatus venom shares 82% sequence identity with imperatoxin A. Both scorpion toxins are putative mimics of the II-III loop peptide (termed peptide A (pA)) of alpha(1s)-dihydropyridine receptor and are thought to act at a common site on ryanodine receptor type 1 (RyR1) important for skeletal muscle EC coupling. The relationship between the actions of synthetic MCa (sMCa) and pA on RyR1 were examined. sMCa released Ca(2+) from SR vesicles (EC(50) = 17.5 nm) in a manner inhibited by micromolar ryanodine or ruthenium red. pA (0.5-40 microm) failed to induce SR Ca(2+) release. Rather, pA enhanced Ca(2+) loading into SR and fully inhibited Ca(2+)-, caffeine-, and sMCa-induced Ca(2+) release. The two peptides modified single channel gating behavior in distinct ways. With Cs(+)-carrying current, 10 nm to 1 microm sMCa induced long lived subconductances having 48% of the characteristic full open state and occasional transitions to 29% at either positive or negative holding potentials. In contrast, pA stabilized long lived channel closures with occasional burst transitions to 65% (s1) and 86% (s2) of the full conductance. The actions of pA and sMCa were observed in tandem. sMCa stabilized additional subconductance states proportional to pA-induced subconductances (i.e. 43% of pA modified s1 and s2 substates), revealing a proportional gating mechanism. [(3)H]Ryanodine binding and surface plasmon resonance analyses indicated that the peptides did not interact by simple competition for a single class of mutually exclusive sites on RyR1 to produce proportional gating. The actions of sMCa were also observed with ryanodine-modified channels and channels deficient in immunophilin 12-kDa FK506-binding protein. These results provide evidence that sMCa and pA stabilize distinct RyR1 channel states through distinct mechanisms that allosterically stabilize gating states having proportional conductance. PMID- 12586832 TI - Calmodulin oxidation and methionine to glutamine substitutions reveal methionine residues critical for functional interaction with ryanodine receptor-1. AB - Calmodulin (CaM) binds to the skeletal muscle ryanodine receptor Ca(2+) release channel (RyR1) with high affinity, and it may act as a Ca(2+)-sensing subunit of the channel. Apo-CaM increases RyR1 channel activity, but Ca(2+)-CaM is inhibitory. Here we examine the functional effects of CaM oxidation on RyR1 regulation by both apo-CaM and Ca(2+)-CaM, as assessed via determinations of [(3)H]ryanodine and [(35)S]CaM binding to skeletal muscle sarcoplasmic reticulum vesicles. Oxidation of all nine CaM Met residues abolished functional interactions of CaM with RyR1. Incomplete CaM oxidation, affecting 5-8 Met residues, increased the CaM concentration required to modulate RyR1, having a greater effect on the apo-CaM species. Mutating individual CaM Met residues to Gln demonstrated that Met-109 was required for apo-CaM activation of RyR1 but not for Ca(2+)-CaM inhibition of the channel. Furthermore, substitution of Gln for Met-124 increased the apo- and Ca(2+)-CaM concentrations required to regulate RyR1. These results thus identify Met residues critical for the productive association of CaM with RyR1 channels and suggest that oxidation of CaM may contribute to altered regulation of sarcoplasmic reticulum Ca(2+) release during oxidative stress. PMID- 12586833 TI - The lysophospholipid receptor G2A activates a specific combination of G proteins and promotes apoptosis. AB - G2A, a G protein-coupled receptor for which lysophosphatidylcholine (LPC) is a high affinity ligand, belongs to a newly defined lysophospholipid receptor subfamily. Expression of G2A is transcriptionally up-regulated by stress-inducing and cell-damaging agents, and ectopic expression of G2A leads to growth inhibition. However, the G proteins that functionally couple to G2A have not been elucidated in detail. We report here that G2A ligand independently stimulates the accumulation of both inositol phosphates and cAMP. LPC does not further enhance inositol phosphate accumulation but dose-dependently augments intracellular cAMP concentration. Expression of G alpha(q) and G alpha(13) with G2A potentiates G2A mediated activation of a NF-kappa B-luciferase reporter. These results demonstrate that G2A differentially couples to multiple G proteins including G alpha(s), G alpha(q), and G alpha(13), depending on whether it is bound to ligand. G2A-transfected HeLa cells display apoptotic signs including membrane blebbing, nuclear condensation, and reduction of mitochondrial membrane potential. Furthermore, G2A-induced apoptosis can be rescued by the caspase inhibitors, z-vad-fmk and CrmA. Although apoptosis occurs without LPC stimulation, LPC further enhances G2A-mediated apoptosis and correlates with its ability to induce cAMP elevation in both HeLa cells and primary lymphocytes. Rescue from G2A-induced apoptosis was achieved by co-expression of a G alpha(12/13)-specific inhibitor, p115RGS (regulator of G protein signaling), in combination with 2',5'-dideoxyadenosine treatment. These results demonstrate the ability of G2A to activate a specific combination of G proteins, and that G2A/LPC induced apoptosis involves both G alpha(13)- and G alpha(s)-mediated pathways. PMID- 12586834 TI - Complementary impact of paralogous Oxa1-like proteins of Bacillus subtilis on post-translocational stages in protein secretion. AB - In mitochondria, chloroplasts, and Gram-negative eubacteria, Oxa1p(-like) proteins are critical for the biogenesis of membrane proteins. Here we show that the Gram-positive eubacterium Bacillus subtilis contains two functional Oxa1p orthologues, denoted SpoIIIJ and YqjG. The presence of either SpoIIIJ or YqjG is required for cell viability. Whereas SpoIIIJ is required for sporulation, YqjG is dispensable for this developmental process. The stability of two membrane proteins was found to be mildly affected upon SpoIIIJ limitation in the absence of YqjG. Surprisingly, the topology and stability of other membrane proteins remained unaffected under these conditions. In contrast, SpoIIIJ- and YqjG limiting conditions resulted in a strong post-translocational defect in the stability of secretory proteins. Together, these data indicate that SpoIIIJ and YqjG of B. subtilis are involved in both membrane protein biogenesis and protein secretion. However, the reduced stability of secretory proteins seems to be the most prominent phenotype of SpoIIIJ/YqjG-depleted B. subtilis cells. In conclusion, our observations show that SpoIIIJ and YqjG have different, but overlapping functions in B. subtilis. Most importantly, it seems that different members of the Oxa1p protein family have acquired at least partly distinct, species-specific, functions that are essential for life. PMID- 12586835 TI - Mitotic regulation of ribosomal S6 kinase 1 involves Ser/Thr, Pro phosphorylation of consensus and non-consensus sites by Cdc2. AB - During mitosis, the cyclin-dependent kinase, Cdc2, signals the inactivation of major anabolic processes such as transcription, mRNA processing, translation, and ribosome biogenesis, thereby providing energy needed for the radical and energetically costly structural reorganization of the cell. This is accomplished by phosphorylation and inactivation of several key anabolic elements, including TFIIIB, TFIID, RNA polymerase II, poly(A) polymerase, and translation elongation factor 1gamma. We report here that ribosomal S6 kinase 1 (S6K1), a protein kinase linked to the translation of ribosomal protein mRNAs, is also subject to regulation by Cdc2 in mitosis. In mitotic HeLa cells, when the activity of Cdc2 is high, S6K1 is phosphorylated at multiple Ser/Thr, Pro (S/TP) sites, including Ser(371), Ser(411), Thr(421), and Ser(424). Concomitant with this, the phosphorylation of the hydrophobic motif site, Thr(389), is reduced resulting in a decrease in the specific activity of S6K1. The mitotic S/TP phosphorylation sites are readily phosphorylated by Cdc2.cyclin B in vitro. These proline directed phosphorylations are sensitive to chemical inhibitors of Cdc2 but not to inhibitors of mammalian target of rapamycin, phosphatidylinositol 3-kinase, MEK1/2, or p38. In murine FT210 cells arrested in mitosis, conditional inactivation of Cdc2 reduces phosphorylation of S6K1 at S/TP sites while simultaneously increasing phosphorylation of Thr(389) and of the S6K1 substrate, RPS6. A physical interaction exists between Cdc2 and S6K1, and this interaction is enhanced in mitotic cells. These results suggest that Cdc2 provides a signal that triggers inactivation of S6K1 in mitosis, presumably serving to spare energy for costly mitotic processes at the expense of ribosomal protein synthesis. PMID- 12586836 TI - Identification of a region of beta-amyloid precursor protein essential for its gelatinase A inhibitory activity. AB - Because beta-amyloid precursor protein (APP) has the abilities both to interact with extracellular matrix and to inhibit gelatinase A activity, this molecule is assumed to play a regulatory role in the gelatinase A-catalyzed degradation of extracellular matrix. To determine a region of APP essential for the inhibitory activity, we prepared various derivatives of APP. Functional analyses of proteolytic fragments of soluble APP (sAPP) and glutathione S-transferase fusion proteins, which contain various COOH-terminal parts of sAPP, showed that a site containing residues 579-601 of APP(770) is essential for the inhibitory activity. Moreover, a synthetic decapeptide containing the ISYGNDALMP sequence corresponding to residues 586-595 of APP(770) had a gelatinase A inhibitory activity slightly higher than that of sAPP. Studies of deletion of the NH(2)- and COOH-terminal residues and alanine replacement of internal residues of the decapeptide further revealed that Tyr(588), Asp(591), and Leu(593) of APP mainly stabilize the interaction between gelatinase A and the inhibitor. We also found that the residues of Ile(586), Met(594), and Pro(595) modestly contribute to the inhibitory activity. The APP-derived decapeptide efficiently inhibited the activity of gelatinase A (IC(50) = 30 nm), whereas its inhibitory activity toward membrane type 1 matrix metalloproteinase was much weaker (IC(50) = 2 microm). The decapeptide had poor inhibitory activity toward gelatinase B, matrilysin, and stromelysin (IC(50) > 10 microm). The APP-derived inhibitor formed a complex with active gelatinase A but not with progelatinase A, and the complex formation was prevented completely by a hydroxamate-based synthetic inhibitor. Therefore, the decapeptide region of APP is likely an active site-directed inhibitor that has high selectivity toward gelatinase A. PMID- 12586837 TI - Activation of pro-gelatinase B by endometase/matrilysin-2 promotes invasion of human prostate cancer cells. AB - This work has explored a putative biochemical mechanism by which endometase/matrilysin-2/matrix metalloproteinase-26 (MMP-26) may promote human prostate cancer cell invasion. Here, we showed that the levels of MMP-26 protein in human prostate carcinomas from multiple patients were significantly higher than those in prostatitis, benign prostate hyperplasia, and normal prostate glandular tissues. The role of MMP-26 in prostate cancer progression is unknown. MMP-26 was capable of activating pro-MMP-9 by cleavage at the Ala(93)-Met(94) site of the prepro-enzyme. This activation proceeded in a time- and dose dependent manner, facilitating the efficient cleavage of fibronectin by MMP-9. The activated MMP-9 products generated by MMP-26 appeared more stable than those cleaved by MMP-7 under the conditions tested. To investigate the contribution of MMP-26 to cancer cell invasion via the activation of MMP-9, highly invasive and metastatic human prostate carcinoma cells, androgen-repressed prostate cancer (ARCaP) cells were selected as a working model. ARCaP cells express both MMP-26 and MMP-9. Specific anti-MMP-26 and anti-MMP-9 functional blocking antibodies both reduced the invasiveness of ARCaP cells across fibronectin or type IV collagen. Furthermore, the introduction of MMP-26 antisense cDNA into ARCaP cells significantly reduced the MMP-26 protein level in these cells and strongly suppressed the invasiveness of ARCaP cells. Double immunofluorescence staining and confocal laser scanning microscopic images revealed that MMP-26 and MMP-9 were co-localized in parental and MMP-26 sense-transfected ARCaP cells. Moreover, MMP-26 and MMP-9 proteins were both expressed in the same human prostate carcinoma tissue samples examined. These results indicate that MMP-26 may be a physiological and pathological activator of pro-MMP-9. PMID- 12586838 TI - Identification of phospholipid scramblase 1 as a novel interacting molecule with beta -secretase (beta -site amyloid precursor protein (APP) cleaving enzyme (BACE)). AB - beta-Site amyloid precursor protein (APP)-cleaving enzyme (BACE) is an integral membrane aspartic proteinase responsible for beta-site processing of APP, and its cytoplasmic region composed of 24 amino acid residues has been shown to be involved in the endosomal localization of BACE. With the yeast two-hybrid screening, we found that the cytoplasmic domain of phospholipid scramblase 1 (PLSCR1), a type II integral membrane protein, interacts with the cytoplasmic region of BACE. In cultured cells, BACE and PLSCR1 were colocalized in the Golgi area and in endosomal compartments, whereas they were co-redistributed in late endosome-derived multivesicular bodies when treated with U18666A, suggesting that both proteins share a common trafficking pathway in cells. Co-immunoprecipitation analysis showed that both proteins form a protein complex at an endogenous expression level in the human neuroblastoma SH-SY5Ycells, and the dileucine residue of the BACE tail is also revealed to be essential for the physical interaction with PLSCR1 in vitro and in vivo. Moreover, both BACE and PLSCR1 were localized in a low buoyant lipid microdomain in SH-SY5Y cells. The dileucine defective BACE mutant was also fractionated into the lipid microdomain, but much less stably than wild-type BACE. Taken together, our current study suggests the functional involvement of PLSCR1 in the intracellular distribution of BACE and/or recruitment of BACE into the detergent-insoluble lipid raft. PMID- 12586839 TI - Phosphorylation motifs regulating the stability and function of myocyte enhancer factor 2A. AB - The phosphorylation status of the myocyte enhancer factor 2 (MEF2) transcriptional regulator is a critical determinant of its tissue-specific functions. However, due to the complexity of its phosphorylation pattern in vivo, a systematic inventory of MEF2A phosphorylation sites in mammalian cells has been difficult to obtain. We employed modern affinity purification techniques, combined with mass spectrometry, to identify several novel MEF2 phosphoacceptor sites. These include an evolutionarily conserved KSP motif, which we show is important in regulating the stability and function of MEF2A. Also, an indirect pathway in which a protein kinase casein kinase 2 phosphoacceptor site is phosphorylated by activation of p38 MAPK signaling was documented. Together, these findings identify several novel aspects of MEF2 regulation that may prove important in the control of gene expression in neuronal and muscle cells. PMID- 12586840 TI - Physical and functional interactions among AP-2 transcription factors, p300/CREB binding protein, and CITED2. AB - The transcriptional co-activators and histone acetyltransferases p300/CREB binding protein (CBP) interact with CITED2, a transcription factor AP-2 (TFAP2) co-activator. p300/CBP, CITED2, and TFAP2A are essential for normal neural tube and cardiac development. Here we show that p300 and CBP co-activate TFAP2A in the presence of CITED2. TFAP2A transcriptional activity was modestly impaired in p300(+/-) and CBP(+/-) mouse embryonic fibroblasts; this was rescued by ectopic expression of p300/CBP. p300, TFAP2A, and endogenous CITED2 could be co immunoprecipitated from transfected U2-OS cells indicating that they can interact physically in vivo. CITED2 interacted with the dimerization domain of TFAP2C, which is highly conserved in TFAP2A/B. In mammalian two-hybrid experiments, full length p300 and TFAP2A interacted only when CITED2 was co-transfected. N-terminal residues of TFAP2A, containing the transactivation domain, are both necessary and sufficient for interaction with p300, and this interaction was independent of CITED2. Consistent with this, N-terminal residues of TFAP2A were required for p300- and CITED2-dependent co-activation. A histone acetyltransferase-deficient p300 mutant (D1399Y) did not co-activate TFAP2A and did not affect the expression or cellular localization of TFAP2A or CITED2. In mammalian two-hybrid experiments p300D1399Y failed to interact with TFAP2A, explaining, at least in part, its failure to function as a co-activator. Our results suggest a model wherein interactions among TFAP2A, CITED2, and p300/CBP are necessary for TFAP2A-mediated transcriptional activation and for normal neural tube and cardiac development. PMID- 12586841 TI - Triiodothyronine decreases the activity of the proximal promoter (PII) of the aromatase gene in the mouse Sertoli cell line, TM4. AB - Estrogens and thyroid hormones play a significant role in regulating functions and development of the testis. The synthesis of estrogens from androgens is catalyzed by the enzyme complex termed aromatase, which in the testis displays an age-related cellular compartmentalization, primarily in Sertoli cells in immature animals, whereas in adults it is expressed in Leydig and germ cells. T3 induces a precocious terminal differentiation of prepubertal Sertoli cells together with a dramatic decrease of their aromatase activity. In the present work, we have examined the mechanism by which T3 exerts this inhibitory action on aromatase expression. As an experimental model, we used the mouse Sertoli cell line TM4, which conserves a large spectrum of functional features present in immature Sertoli cells. For instance, after revealing the presence of aromatase by immunocytochemistry and measuring its enzymatic activity, we confirmed in this cell line the functional events previously characterized in primary cultures of immature rat Sertoli cells: 1) a strong stimulation of aromatase activity by dibutyryl-cAMP [(Bu)2cAMP] (simulating FSH action); and 2) the inhibition of aromatase activity by incubation with T3 under basal condition and after (Bu)2cAMP stimulation. After identifying promoter II as the regulatory region located immediately upstream of the transcriptional initiation site in the TM4 cell line by rapid amplification of cDNA ends analysis, we conducted experiments to examine the molecular mechanism by which thyroid hormones modulate aromatase gene expression in this cell line. TM4 cells were transfected with plasmids containing different segments of the rat promoter II sequence ligated to a luciferase reporter gene. Analysis of the activities of these promoter fusions demonstrated that T3 inhibits basal and (Bu)2cAMP-stimulated activity of the aromatase promoter. This effect was not revealed in T3-treated cells transfected with construct in which the steroidogenic factor-1 (SF-1) response element was mutated. These results indicate that the inhibitory effect of T3 requires the integrity of the SF-1 response element and are further supported in the EMSA. The EMSA experiments demonstrated that thyroid hormone/thyroid receptor alpha1 complex (TH/TRalpha1) is able to compete with SF-1 in binding to oligonucleotides containing an SF-1 motif, an element essential for the activity of the PII aromatase promoter. The findings suggest that the binding of the thyroid hormone/thyroid receptor alpha1 complex to the SF-1 motif is the molecular mechanism by which T3 exerts an inhibitory effect on aromatase gene expression in the TM4 cell line. PMID- 12586842 TI - Transcriptional activation by thyroid hormone receptor-beta involves chromatin remodeling, histone acetylation, and synergistic stimulation by p300 and steroid receptor coactivators. AB - Transcriptional regulation by heterodimers of thyroid hormone receptor (TR) and the 9-cis retinoid X receptor (RXR) is a highly complex process involving a large number of accessory factors, as well as chromatin remodeling. We have used a biochemical approach, including an in vitro chromatin assembly and transcription system that accurately recapitulates ligand- and activation function (AF)-2 dependent transcriptional activation by TRbeta/RXRalpha heterodimers, as well as in vitro chromatin immunoprecipitation assays, to study the mechanisms of TRbeta mediated transcription with chromatin templates. Using this approach, we show that chromatin is required for robust ligand-dependent activation by TRbeta. We also show that the binding of liganded TRbeta to chromatin induces promoter proximal chromatin remodeling and histone acetylation, and that histone acetylation is correlated with increased TRbeta-dependent transcription. Additionally, we find that steroid receptor coactivators (SRCs) and p300 function synergistically to stimulate TRbeta-dependent transcription, with multiple functional domains of p300 contributing to its coactivator activity with TRbeta. A major conclusion from our experiments is that the primary role of the SRC proteins is to recruit p300/cAMP response element binding protein-binding protein to hormone-regulated promoters. Together, our results suggest a multiple step pathway for transcriptional regulation by liganded TRbeta, including chromatin remodeling, recruitment of coactivators, targeted histone acetylation, and recruitment of the RNA polymerase II transcriptional machinery. Our studies highlight the functional importance of chromatin in transcriptional control and further define the molecular mechanisms by which the SRC and p300 coactivators facilitate transcriptional activation by liganded TRbeta. PMID- 12586843 TI - A novel antiinflammatory maintains glucocorticoid efficacy with reduced side effects. AB - Glucocorticoids (GCs) are commonly used to treat inflammatory disease; unfortunately, the long-term use of these steroids leads to a large number of debilitating side effects. The antiinflammatory effects of GCs are a result of GC receptor (GR)-mediated inhibition of expression of proinflammatory genes as well as GR-mediated activation of antiinflammatory genes. Similarly, side effects are most likely due to both activated and repressed GR target genes in affected tissues. An as yet unachieved pharmaceutical goal is the development of a compound capable of separating detrimental side effects from antiinflammatory activity. We describe the discovery and characterization of AL-438, a GR ligand that exhibits an altered gene regulation profile, able to repress and activate only a subset of the genes normally regulated by GCs. When tested in vivo, AL-438 retains full antiinflammatory efficacy and potency comparable to steroids but its negative effects on bone metabolism and glucose control are reduced at equivalently antiinflammatory doses. The mechanism underlying this selective in vitro and in vivo activity may be the result of differential cofactor recruitment in response to ligand. AL-438 reduces the interaction between GR and peroxisomal proliferator-activated receptor gamma coactivator-1, a cofactor critical for steroid-mediated glucose up-regulation, while maintaining normal interactions with GR-interacting protein 1. This compound serves as a prototype for a unique, nonsteroidal alternative to conventional GCs in treating inflammatory disease. PMID- 12586844 TI - Signal transducer and activator of transcription 5 activation is sufficient to drive transcriptional induction of cyclin D2 gene and proliferation of rat pancreatic beta-cells. AB - Signal transducer and activator of transcription 5 (STAT5) activation plays a central role in GH- and prolactin-mediated signal transduction in the pancreatic beta-cells. In previous experiments we demonstrated that STAT5 activation is necessary for human (h)GH-stimulated proliferation of INS-1 cells and hGH-induced increase of mRNA-levels of the cell cycle regulator cyclin D2. In this study we have further characterized the role of STAT5 in the regulation of cyclin D expression and beta-cell proliferation by hGH. Cyclin D2 mRNA and protein levels (but not cyclin D1 and D3) were induced in a time-dependent manner by hGH in INS 1 cells. Inhibition of protein synthesis by coincubation with cycloheximide did not affect the hGH-induced increase of cyclin D2 mRNA levels at 4 h. Expression of a dominant negative STAT5 mutant, STAT5aDelta749, partially inhibited cyclin D2 protein levels. INS-1 cells transiently transfected with a cyclin D2 promoter reporter construct revealed a 3- to 5-fold increase of transcriptional activity in response to hGH stimulation. Furthermore, coexpression of a constitutive active STAT5 mutant (either CA-STAT5a or CA-STAT5b) was sufficient to drive transactivation of the promoter. CA-STAT5b was stably expressed in INS-1 cells under the control of a doxycycline-inducible promoter. Gel retardation experiments using a probe representing a putative STAT5 binding site in the cyclin D2 promoter revealed binding of the doxycycline-induced CA-STAT5b. Furthermore, induction of CA-STAT5b stimulated transcriptional activation of the cyclin D2 promoter and induced hGH-independent proliferation in these cells. In primary beta-cells, adenovirus-mediated expression of CA-STAT5b profoundly stimulated DNA-synthesis (5.3-fold over control) in the absence of hGH. Our studies indicate that STAT5 activation is sufficient to drive proliferation of the beta-cells and that cyclin D2 may be a critical target gene for STAT5 in this process. PMID- 12586845 TI - Stromal cell-derived factor 1, a novel target of estrogen receptor action, mediates the mitogenic effects of estradiol in ovarian and breast cancer cells. AB - Recent clinical studies estimate that 60-70% of human ovarian and breast cancers overexpress the estrogen receptor (ER). However, despite the established mitogenic effects of estrogen in these tumors, proliferative markers of hormone action are limited. In the current study, we report that the growth stimulatory cytokine stromal cell-derived factor 1 (SDF-1) is a bona fide target of estrogen action in ERalpha-positive human ovarian and breast cancer cells. Notably, estradiol treatment of BG-1 (ovarian carcinoma) and MCF-7 (breast carcinoma) cells leads to rapid and robust induction of the SDF-1alpha and beta isoforms. This response is blocked by the pure ER antagonist ICI 182,780 and is not apparent in ER-negative ovarian cells, indicating that SDF-1 regulation is ERalpha mediated. Treatment with the protein synthesis inhibitor cycloheximide had no effect on estradiol induction of induction of SDF-1 mRNA levels mRNA levels, demonstrating that SDF-1 is a direct target of ERalpha. SDF-1 protein levels, although undetectable under basal conditions, were strikingly increased by hormone both intracellularly and in the media of cultured BG-1 and MCF-7 cells. In cell proliferation assays, the mitogenic effects of estradiol were neutralized by addition of an SDF-1 antibody and mimicked by the addition of exogenous SDF-1 protein, indicating that SDF-1 mediates the proliferative actions of hormone. Furthermore, activation of the SDF-1 receptor CXCR4 stimulated BG-1 and MCF-7 cell proliferation in a manner comparable to estradiol. Taken together, these results demonstrate a novel estrogen-mediated paracrine pathway for inducing cancer cell proliferation and suggest that SDF-1 and CXCR4 may represent novel therapeutic targets in ERalpha-positive ovarian and breast tumors. PMID- 12586846 TI - Robustness into advanced age of atopy-specific mechanisms in atopy-prone families. AB - We evaluated atopy-associated parameters in 1,099 people (aged 6-84 years) from families with history for atopy. All were tested for serum total immunoglobulin E (IgE) and allergen sensitivity by skin prick test. Specific IgE tests were done in randomly selected families. There was a decline with age in serum total IgE values, and relative atopy "incidence rates" were slightly lower among those older than 60 years. However, there was no change with age in sensitivity or severity of atopy. Among those sensitized to ragweed (Ambrosia artemisilfolia), there was no age-associated change in IgE levels specific to Amb a 1, a major allergen extracted from ragweed, and no change in the binding affinity of IgE for the Amb a 1 allergen. Among families with atopic histories, the underlying atopic mechanisms are particularly robust, and the atopic propensity remains into advanced age. In addition, established atopic responses may be focused in an immune system compartment either independent of or minimally influenced by T-cell activity. PMID- 12586847 TI - Gender differences in resistance-training-induced myofiber hypertrophy among older adults. AB - We tested the hypothesis that older men (n = 9, 69 +/- 2 years) would experience greater resistance-training-induced myofiber hypertrophy than older women (n = 5, 66 +/- 1 years) following knee extensor training 3 days per week at 65-80% of one repetition maximum for 26 weeks. Vastus lateralis biopsies were analyzed for myofiber areas, myosin heavy chain isoform distribution, and levels of mRNA for insulin-like growth factor 1 (IGF-1), IGFR1, and myogenin. Gender x Training interactions (p <.05) indicate greater myofiber hypertrophy for all three primary fiber types (I, IIa, IIx) and enhanced one-repetition maximum strength gain in men compared with women (p <.05). Covarying for serum IGF-1, dehydroepiandrosterone sulfate, or each muscle mRNA did not negate these interactions. In both genders, type IIx myofiber area distribution and myosin heavy chain type IIx distribution decreased with a concomitant increase in type IIa myofiber area distribution (p <.05). In summary, gender differences in load induced myofiber hypertrophy among older adults cannot be explained by levels of circulating IGF-1 or dehydroepiandrosterone sulfate, or by expression of the myogenic transcripts examined. PMID- 12586848 TI - Different glial reactions to hippocampal stab wounds in young adult and aged rats. AB - Brain injury induces reactive gliosis. To examine the activation of glial cells after brain injury in young versus aged rats, we used a brain stab-wound model and examined the expression of cells positive for ED1 (ED1(+)) and glial fibrillary acidic protein (GFAP(+)) in the hippocampus in young-mature (3 months) and aged (25 months) Wistar rats at various times following hippocampal stab injury. ED1(+) cells appeared more frequently in the aged rats than in the young mature rats under control conditions, whereas the number of GFAP(+) cells was not different between two groups. Following the stab wound, there was an increase in ED1 expression that was delayed but stronger in the aged rats and that persisted longer; the increase of the number of GFAP(+) cells also persisted longer. We conclude that different glial reactivity in the aged brain suggests that aging is associated with increased glial responsiveness that may enhance susceptibility to injury and disease in the brain. PMID- 12586849 TI - Increased cross-sectional area and reduced tensile stress of the Achilles tendon in elderly compared with young women. AB - The Achilles tendon cross-sectional area (CSA), tensile force, and stress during an isometric contraction were examined in healthy young (n = 9, age = 29 +/- 1 years, mean +/- SEM) and elderly (n = 10, 79 +/- 2 years) women. CSA area was obtained with magnetic resonance imaging 3 cm proximal to the insertion, and tendon force was obtained from the isometric ankle moment. The moment of force about the ankle joint was greater in young women (95 +/- 17 N m) than in elderly women (51 +/- 5 N m; p <.05). The Achilles tendon CSA was significantly greater in elderly women (56.3 +/- 3.0 mm(2)) than in young women (46.0 +/- 1.9 mm(2); p <.01). These data show that young women can exert a greater force than elderly women on the Achilles tendon during voluntary contraction, although elderly women have an increased (22%) tendon CSA, and a lower tendon force than young women. The greater tendon size combines to lower the stress on the tendon markedly, which may reduce the risk of injury to the tendon. PMID- 12586850 TI - Anorexia and weight loss in older persons. PMID- 12586851 TI - The aging-disease dichotomy: true or false? PMID- 12586852 TI - The aging hand. AB - Hand function decreases with age in both men and women, especially after the age of 65 years. A review is presented of anatomical and physiological changes in the aging hand. The age-related changes in prehension patterns (grip and pinch strength) and hand dexterity in the elderly population are considered. Deterioration in hand function in the elderly population is, to a large degree, secondary to age-related degenerative changes in the musculoskeletal, vascular, and nervous systems. Deterioration of hand function in elderly adults is a combination of local structural changes (joints, muscle, tendon, bone, nerve and receptors, blood supply, skin, and fingernails) and more distant changes in neural control. These age-related changes are often accompanied by underlying pathological conditions (osteoporosis, osteoarthritis, rheumatic arthritis, and Parkinson's disease) that are common in the elderly population. Assessment of hand function and prehension patterns is needed in order to determine specific treatment approaches. PMID- 12586853 TI - Direct and indirect effects of everyday emotions on food intake of elderly patients in institutions. AB - BACKGROUND: Decreased food intake is an important risk factor for malnutrition, which is highly prevalent among geriatric patients. The emotional nature of the hospitalization experience and the complex organizational setting involved in meal production and delivery services in institutions increase the risk for decreased food intake. Everyday emotions are known to have a particularly strong influence on decision-making and behavior in the elderly, and have also been shown, in younger populations, to influence food intake and its psychological antecedents, such as quality perception and satisfaction judgments. The objective of this paper is to study the direct impact of elderly patients' everyday emotions on food intake and their indirect effects mediated by quality perceptions and satisfaction judgments. METHODS: Thirty patients (20 women, 10 men, 65-92 age range) in a geriatric rehabilitation unit were observed on repeated meal episodes (average of 46 care episodes per patient) where they provided self-reports for emotions (positive emotions, anger, anxiety, and mild depressed feelings), perceived meal quality, and satisfaction. Food intake was measured in terms of energy and protein content. RESULTS: The impact on food intake was favorable, and both direct and indirect for positive emotions, direct and negative for anxiety, direct and positive for mild depressed feelings, and indirect and negative for anger. Indirect effects were mediated by quality perception judgments but not by satisfaction, which was not significantly related to food intake. CONCLUSION: Results suggest that, given their impact on food intake, measuring and monitoring patients' everyday emotions may be an important innovative strategy to improve food intake of elderly patients in institutions. PMID- 12586854 TI - The influence of protein-calorie malnutrition on quality of life in nursing homes. AB - BACKGROUND: Up to 85% of the older adults living in our nation's nursing homes suffer from protein-calorie malnutrition (PCM). Early identification and treatment of PCM can reduce or prevent hospital stays, reduce complications, and decrease mortality. We describe the influence of PCM on quality of life in nursing homes, using archived data from the Minimum Data Set. METHODS: The study was guided by the Quality Nutrition Outcomes-Long Term Care Model, which posits a pathway whereby organizational issues influence nutritional status, consisting of body mass index (BMI), serum albumin levels, and prealbumin levels, and subsequent quality of life, morbidity, and health care utilization. A cross sectional design was used to analyze Minimum Data Set assessment data already collected from a previous study. The sample for this analysis was 311 nursing home residents, aged 65 years or older, who lived in three nursing homes in eastern Washington. RESULTS: Of the participants, 38.6% were malnourished. PCM (measured by BMI) influenced quality of life for these residents in that there was a significant relationship between BMI and functional status (eating, personal hygiene, and toilet use) and BMI and psychosocial well-being (initiative or involvement, unsettled relationships, and past roles). Depression was not a significant indicator of low BMI in these nursing home residents. CONCLUSIONS: Low BMI, indicating PCM, was found to negatively influence quality of life in this study. Understanding the relationship between quality of life and PCM could lead to improved quality of life for older adults in nursing homes and guide future innovative intervention studies aimed at preventing PCM. PMID- 12586855 TI - Effects of testosterone replacement and/or resistance training on interleukin-6, tumor necrosis factor alpha, and leptin in elderly men ingesting megestrol acetate: a randomized controlled trial. AB - BACKGROUND: Megestrol acetate (MA) has been used to stimulate weight gain in elderly populations with the majority of the gain being adipose tissue. Significant inverse correlations have been reported between weight gain with MA and reductions in circulating the tumor necrosis factor (TNF) alpha receptors. In addition, MA has been shown to reduce circulating interleukin 6 (IL-6) concentrations. We attempted to increase gains in fat-free mass with MA using resistance training and/or testosterone replacement and examined the effects on circulating IL-6, TNF alpha, and leptin in elderly men. METHODS: All subjects received MA and were randomly assigned to one of these four groups: placebo (P) injections; resistance training and P (RT+P); weekly injections of testosterone (T; 100 mg/wk); or RT and T (RT+T). Cytokines were measured by enzyme-linked immunosorbent assay Preinterventions, Midinterventions, and after 12 weeks of the interventions (Post). RESULTS: IL-6 decreased ( p =.03) over time for the T and P groups when compared to the RT+T and RT+P groups. A time effect ( p =.013) was observed for TNF alpha with Mid and Post both being lower than Pre. A hormone by time interaction ( p =.03) was observed for plasma leptin with individuals not on T exhibiting higher concentrations Post than those individuals on T. A positive correlation was observed (r =.60; p <.05) between changes in fat mass and the change in leptin. CONCLUSION: IL-6 was reduced by MA except when RT was undertaken; TNF alpha was reduced over time regardless of group; leptin was higher in individuals not on T than those on T; and the change in plasma leptin was correlated with the change in fat mass. PMID- 12586856 TI - Effect of strength and power training on physical function in community-dwelling older adults. AB - BACKGROUND: The performance of daily tasks, such as stair climbing or lifting an object, requires both muscle strength and power. Age-associated reductions in strength and power can affect an older adult's ability to complete daily tasks such as stair climbing and lifting a child. METHODS: The purposes of this study were to determine whether power training was more efficacious than strength training for improving whole-body physical function in older adults and to examine the relationship between changes in anaerobic power and muscle strength and changes in physical function. Thirty-nine men and women (mean age +/- SD = 72.5 +/- 6.3 years) with below-average leg extensor power were randomly assigned to control (C, n = 15), strength-training (ST, n = 13) or power-training (PT, n = 11) groups. The ST and PT groups met 3 days per week for 16 weeks; the C group maintained usual activity and attended three lectures during the course of the study. Primary outcome measures included the Continuous Scale Physical Functional Performance test, maximal strength, and anaerobic power. RESULTS: After baseline was controlled for, the Continuous Scale Physical Functional Performance test total score was significantly greater for the PT group than for the ST (p =.033) and C (p =.016) groups. Maximal strength was significantly greater for the ST group than for the C group (p =.015) after the intervention. There was no significant difference between groups for peak anaerobic power. CONCLUSIONS: Power training was more effective than strength training for improving physical function in community-dwelling older adults. PMID- 12586857 TI - Aerobic fitness reduces brain tissue loss in aging humans. AB - BACKGROUND: The human brain gradually loses tissue from the third decade of life onward, with concomitant declines in cognitive performance. Given the projected rapid growth in aged populations, and the staggering costs associated with geriatric care, identifying mechanisms that may reduce or reverse cerebral deterioration is rapidly emerging as an important public health goal. Previous research has demonstrated that aerobic fitness training improves cognitive function in older adults and can improve brain health in aging laboratory animals, suggesting that aerobic fitness may provide a mechanism to improve cerebral health in aging humans. We examined the relationship between aerobic fitness and in vivo brain tissue density in an older adult population, using voxel-based morphometric techniques. METHODS: We acquired high-resolution magnetic resonance imaging scans from 55 older adults. These images were segmented into gray and white matter maps, registered into stereotaxic space, and examined for systematic variation in tissue density as a function of age, aerobic fitness, and a number of other health markers. RESULTS: Consistent with previous studies of aging and brain volume, we found robust declines in tissue densities as a function of age in the frontal, parietal, and temporal cortices. More importantly, we found that losses in these areas were substantially reduced as a function of cardiovascular fitness, even when we statistically controlled for other moderator variables. CONCLUSIONS: These findings extend the scope of beneficial effects of aerobic exercise beyond cardiovascular health, and they suggest a strong solid biological basis for the benefits of exercise on the brain health of older adults. PMID- 12586858 TI - Lifestyle intervention of hypocaloric dieting and walking reduces abdominal obesity and improves coronary heart disease risk factors in obese, postmenopausal, African-American and Caucasian women. AB - BACKGROUND: There are few empirical data to support the claim that weight loss improves coronary heart disease (CHD) risk factors in postmenopausal women; nor is it known if there are racial differences in changes of body fat distribution, lipids, glucose tolerance, and blood pressure with weight loss. This study determined the efficacy of a lifestyle weight loss intervention in reducing total and abdominal obesity and improving CHD risk factors in obese Caucasian and African-American postmenopausal women. METHODS: Body composition (dual-energy x ray absorptiometry), abdominal fat areas (computed tomography scan), lipoprotein lipids, insulin, glucose tolerance, and blood pressure were measured before and after 6 months of hypocaloric diet and low-intensity walking in 76 overweight or obese (body mass index > 25 kg/m(2)), Caucasian (72%) or African-American (28%), postmenopausal (age = 60 +/- 5 years) women who completed the study. RESULTS: Absolute amount of body weight lost was similar in Caucasians (-5.4 +/- 3.6 kg) and African Americans (-3.9 +/- 3.6 kg), but Caucasian women lost relatively more fat mass (p <.05). Both groups decreased their subcutaneous abdominal fat, and Caucasian women decreased their visceral fat area, but there were no racial differences in the magnitude of abdominal fat lost. The intervention decreased triglyceride and increased high-density lipoprotein and high-density lipoprotein 2 cholesterol in both races, and it decreased total and low-density lipoprotein cholesterol in Caucasian women (p <.05-.0001). Fasting glucose and glucose area during the oral glucose tolerance test decreased (p <.0001) in Caucasian women, whereas insulin area decreased in both Caucasian (p <.01) and African-American (p <.05) women. Blood pressure decreased the most in women with higher blood pressures at baseline. Changes in lipids, fasting glucose and insulin, their responses during the oral glucose tolerance test, and blood pressure were not different between racial groups. CONCLUSIONS: Weight loss achieved through a lifestyle intervention of energy restriction and increased physical activity is an equally effective therapy in African-American and Caucasian obese, postmenopausal women for improving glucose and lipid CHD risk factors. PMID- 12586859 TI - The network for elderly care in Italy: only a correct use of acute wards allows an overall functioning of the health care system. PMID- 12586861 TI - Development of murine ischemic cardiomyopathy is associated with a transient inflammatory reaction and depends on reactive oxygen species. AB - We examined the effects of daily repetitive brief (15 min) myocardial ischemia and reperfusion (I/R) in WT C57BL6 and extracellular superoxide dismutase (EC SOD)-overexpressing mice. In the absence of myocardial necrosis, I/R resulted in persistent fibrosis in ischemic areas of C57/BL6 mice associated with persistent global and segmental anterior wall dysfunction. The I/R protocol induced chemokines (peak 3 days) followed sequentially by infiltration of macrophages and myofibroblasts (5 days). Fibrosis peaked at 7 days and was stable at 28 days despite regression of the chemokine and cellular response. Discontinuation of I/R at 7 or 28 days led to regression of fibrosis and ventricular dysfunction. In contrast, the EC-SOD mice developed markedly less chemokine induction, cell response, and fibrosis, with no ventricular dysfunction. Reversible fibrosis and ventricular dysfunction are features of human hibernating myocardium. The reduction of the cellular and functional response in EC-SOD mice suggests a role for reactive O(2) in the pathogenesis of ischemic cardiomyopathy. PMID- 12586860 TI - Structural origins of adenine-tract bending. AB - DNA sequences containing short adenine tracts are intrinsically curved and play a role in transcriptional regulation. Despite many high-resolution NMR and x-ray studies, the origins of curvature remain disputed. Long-range restraints provided by 85 residual dipolar couplings were measured for a DNA decamer containing an adenine (A)(4)-tract and used to refine the structure. The overall bend in the molecule is a result of in-phase negative roll in the A-tract and positive roll at its 5' junction, as well as positive and negative tilt inside the A-tract and near its junctions. The bend magnitude and direction obtained from NMR structures is 9.0 degrees into the minor groove in a coordinate frame located at the third AT base pair. We evaluated long-range and wedge models for DNA curvature and concluded that our data for A-tract curvature are best explained by a "delocalized bend" model. The global bend magnitude and direction of the NMR structure are in excellent agreement with the junction model parameters used to rationalize gel electrophoretic data and with preliminary results of a cyclization kinetics assay from our laboratory. PMID- 12586862 TI - Control of guard cell ion channels by hydrogen peroxide and abscisic acid indicates their action through alternate signaling pathways. PMID- 12586863 TI - The development of plant leaves. PMID- 12586864 TI - Nitrate finds a place in the sun. PMID- 12586865 TI - Chlamydomonas reinhardtii genome project. A guide to the generation and use of the cDNA information. AB - The National Science Foundation-funded Chlamydomonas reinhardtii genome project involves (a) construction and sequencing of cDNAs isolated from cells exposed to various environmental conditions, (b) construction of a high-density cDNA microarray, (c) generation of genomic contigs that are nucleated around specific physical and genetic markers, (d) generation of a complete chloroplast genome sequence and analyses of chloroplast gene expression, and (e) the creation of a Web-based resource that allows for easy access of the information in a format that can be readily queried. Phases of the project performed by the groups at the Carnegie Institution and Duke University involve the generation of normalized cDNA libraries, sequencing of cDNAs, analysis and assembly of these sequences to generate contigs and a set of predicted unique genes, and the use of this information to construct a high-density DNA microarray. In this paper, we discuss techniques involved in obtaining cDNA end-sequence information and the ways in which this information is assembled and analyzed. Descriptions of protocols for preparing cDNA libraries, assembling cDNA sequences and annotating the sequence information are provided (the reader is directed to Web sites for more detailed descriptions of these methods). We also discuss preliminary results in which the different cDNA libraries are used to identify genes that are potentially differentially expressed. PMID- 12586866 TI - A sequence-based map of Arabidopsis genes with mutant phenotypes. AB - The classical genetic map of Arabidopsis contains 462 genes with mutant phenotypes. Chromosomal locations of these genes have been determined over the past 25 years based on recombination frequencies with visible and molecular markers. The most recent update of the classical map was published in a special genome issue of Science that dealt with Arabidopsis (D.W. Meinke, J.M. Cherry, C. Dean, S.D. Rounsley, M. Koornneef [1998] Science 282: 662-682). We present here a comprehensive list and sequence-based map of 620 cloned genes with mutant phenotypes. This map documents for the first time the exact locations of large numbers of Arabidopsis genes that give a phenotype when disrupted by mutation. Such a community-based physical map should have broad applications in Arabidopsis research and should serve as a replacement for the classical genetic map in the future. Assembling a comprehensive list of genes with a loss-of-function phenotype will also focus attention on essential genes that are not functionally redundant and ultimately contribute to the identification of the minimal gene set required to make a flowering plant. PMID- 12586867 TI - Comparative analyses of potato expressed sequence tag libraries. AB - The cultivated potato (Solanum tuberosum) shares similar biology with other members of the Solanaceae, yet has features unique within the family, such as modified stems (stolons) that develop into edible tubers. To better understand potato biology, we have undertaken a survey of the potato transcriptome using expressed sequence tags (ESTs) from diverse tissues. A total of 61,940 ESTs were generated from aerial tissues, below-ground tissues, and tissues challenged with the late-blight pathogen (Phytophthora infestans). Clustering and assembly of these ESTs resulted in a total of 19,892 unique sequences with 8,741 tentative consensus sequences and 11,151 singleton ESTs. We were able to identify a putative function for 43.7% of these sequences. A number of sequences (48) were expressed throughout the libraries sampled, representing constitutively expressed sequences. Other sequences (13,068, 21%) were uniquely expressed and were detected only in a single library. Using hierarchal and k means clustering of the EST sequences, we were able to correlate changes in gene expression with major physiological events in potato biology. Using pair-wise comparisons of tuber related tissues, we were able to associate genes with tuber initiation, dormancy, and sprouting. We also were able to identify a number of characterized as well as novel sequences that were unique to the incompatible interaction of late-blight pathogen, thereby providing a foundation for further understanding the mechanism of resistance. PMID- 12586868 TI - Gene expression in autumn leaves. AB - Two cDNA libraries were prepared, one from leaves of a field-grown aspen (Populus tremula) tree, harvested just before any visible sign of leaf senescence in the autumn, and one from young but fully expanded leaves of greenhouse-grown aspen (Populus tremula x tremuloides). Expressed sequence tags (ESTs; 5,128 and 4,841, respectively) were obtained from the two libraries. A semiautomatic method of annotation and functional classification of the ESTs, according to a modified Munich Institute of Protein Sequences classification scheme, was developed, utilizing information from three different databases. The patterns of gene expression in the two libraries were strikingly different. In the autumn leaf library, ESTs encoding metallothionein, early light-inducible proteins, and cysteine proteases were most abundant. Clones encoding other proteases and proteins involved in respiration and breakdown of lipids and pigments, as well as stress-related genes, were also well represented. We identified homologs to many known senescence-associated genes, as well as seven different genes encoding cysteine proteases, two encoding aspartic proteases, five encoding metallothioneins, and 35 additional genes that were up-regulated in autumn leaves. We also indirectly estimated the rate of plastid protein synthesis in the autumn leaves to be less that 10% of that in young leaves. PMID- 12586869 TI - Genomic and proteomic analysis of mitochondrial carrier proteins in Arabidopsis. AB - Plant mitochondria maintain metabolic communication with the cytosol through a family of carrier proteins. In Arabidopsis, a subset of 45 putative genes encoding members of this family have been identified based on generalized mitochondrial carrier features. No gene clusters are apparent and few of the predicted protein products have mitochondrial targeting sequences recognized by bioinformatic predictors. Only nine genes are currently represented by more than 10 expressed sequence tags at The Institute for Genomic Research. Analyses of public microarray experiments reveal differential expression profiles of the more highly expressed members of this gene family in different plant organs and in response to plant hormone application and environmental stresses. A comparison of this Arabidopsis carrier subset (45) to the yeast gene family (35) reveals 10 orthologous groups between the two species. Recent surveys of the Arabidopsis mitochondrial proteome by two-dimensional gel separations have not identified any of these carrier proteins, presumably because of their hydrophobicity and basicity. Isolating integral membrane proteins from Arabidopsis mitochondria, using one-dimensional electrophoresis for protein separation and tandem mass spectrometry-based sequencing of doubly charged peptides, we have unequivocally identified specific carrier gene products located in mitochondria. This approach has identified six of the nine carriers represented highly in expressed sequence tag databases: adenine nucleotide translocator (At3g8580 and At5g13490), dicarboxylate/tricarboxylate carrier (At5g19760), phosphate carrier (At5g14040), uncoupling protein (At3g54110), and a carrier gene of unknown function (At4g01100). Overall, the combined transcript and protein expression data indicates that only a small subset of the carrier family of genes provide the majority of carrier proteins of Arabidopsis mitochondria. PMID- 12586870 TI - Osmotic stress tolerance of transgenic tobacco expressing a gene encoding a membrane-located receptor-like protein from tobacco plants. AB - Tobacco (Nicotiana tabacum) genes regulated during the early stage of responses to wounding were screened by a modified fluorescence differential display method. Among 28 genes initially identified, a particular clone designated NtC7 was subjected to further analysis. Its transcripts were found to accumulate rapidly and transiently within 1 h upon treatments with not only wounding but also salt and osmotic stresses. However, jasmonic and abscisic acids and ethylene did not effectively induce NtC7 transcripts. Amino acid sequence analysis suggested NtC7 to be a new type of transmembrane protein that belongs to the receptor-like protein family, and a membrane location was confirmed in onion (Allium cepa) epidermis cells transiently expressing an NtC7-green fluorescent protein fusion protein. Seeds of transgenic tobacco overexpressing NtC7 normally germinated and grew in the presence of 500 mM mannitol, but not in the presence of 220 mM sodium chloride or 60 mM lithium chloride. Cuttings of mature transgenic leaf exhibited a marked tolerance upon treatment with 500 mM mannitol for 12 h, at which concentration wild-type counterparts were seriously damaged. These results suggested that NtC7 predominantly functions in maintenance of osmotic adjustment independently of ion homeostasis. PMID- 12586871 TI - Subcellular targeting of methylmercury lyase enhances its specific activity for organic mercury detoxification in plants. AB - Methylmercury is an environmental pollutant that biomagnifies in the aquatic food chain with severe consequences for humans and other animals. In an effort to remove this toxin in situ, we have been engineering plants that express the bacterial mercury resistance enzymes organomercurial lyase MerB and mercuric ion reductase MerA. In vivo kinetics experiments suggest that the diffusion of hydrophobic organic mercury to MerB limits the rate of the coupled reaction with MerA (Bizily et al., 2000). To optimize reaction kinetics for organic mercury compounds, the merB gene was engineered to target MerB for accumulation in the endoplasmic reticulum and for secretion to the cell wall. Plants expressing the targeted MerB proteins and cytoplasmic MerA are highly resistant to organic mercury and degrade organic mercury at 10 to 70 times higher specific activity than plants with the cytoplasmically distributed wild-type MerB enzyme. MerB protein in endoplasmic reticulum-targeted plants appears to accumulate in large vesicular structures that can be visualized in immunolabeled plant cells. These results suggest that the toxic effects of organic mercury are focused in microenvironments of the secretory pathway, that these hydrophobic compartments provide more favorable reaction conditions for MerB activity, and that moderate increases in targeted MerB expression will lead to significant gains in detoxification. In summary, to maximize phytoremediation efficiency of hydrophobic pollutants in plants, it may be beneficial to target enzymes to specific subcellular environments. PMID- 12586872 TI - Identification of mutants of Arabidopsis defective in acclimation of photosynthesis to the light environment. AB - In common with many other higher plant species, Arabidopsis undergoes photosynthetic acclimation, altering the composition of the photosynthetic apparatus in response to fluctuations in its growth environment. The changes in photosynthetic function that result from acclimation can be detected in a noninvasive manner by monitoring chlorophyll (Chl) fluorescence. This technique has been used to develop a screen that enables the rapid identification of plants defective at ACCLIMATION OF PHOTOSYNTHESIS TO THE ENVIRONMENT (APE) loci. The application of this screen to a population of T-DNA-transformed Arabidopsis has successfully led to the identification of a number of mutant lines with altered Chl fluorescence characteristics. Analysis of photosynthesis and pigment composition in leaves from three such mutants showed that they had altered acclimation responses to the growth light environment, each having a distinct acclimation-defective phenotype, demonstrating that screening for mutants using Chl fluorescence is a viable strategy for the investigation of acclimation. Sequencing of the genomic DNA flanking the T-DNA elements showed that in the ape1 mutant, a gene was disrupted that encodes a protein of unknown function but that appears to be specific to photosynthetic organisms, whereas the ape2 mutant carries an insertion in the region of the TPT gene encoding the chloroplast inner envelope triose phosphate/phosphate translocator. PMID- 12586873 TI - Sequence analysis of a 282-kilobase region surrounding the citrus Tristeza virus resistance gene (Ctv) locus in Poncirus trifoliata L. Raf. AB - Citrus tristeza virus (CTV) is the major virus pathogen causing significant economic damage to citrus worldwide, and a single dominant gene, Ctv, provides broad spectrum resistance to CTV in Poncirus trifoliata L. Raf. Ctv was physically mapped to a 282-kb region using a P. trifoliata bacterial artificial chromosome library. This region was completely sequenced to about 8x coverage using a shotgun sequencing strategy and primer walking for gap closure. Sequence analysis predicts 22 putative genes, two mutator-like transposons and eight retrotransposons. This sequence analysis also revealed some interesting features of this region of the P. trifoliata genome: a disease resistance gene cluster with seven members and eight retrotransposons clustered in a 125-kb gene-poor region. Comparative sequence analysis suggests that six genes in the Ctv region have significant sequence similarity with their orthologs in bacterial artificial chromosome clones F7H2 and F21T11 from Arabidopsis chromosome I. However, the analysis of gene colinearity between P. trifoliata and Arabidopsis indicates that Arabidopsis genome sequence information may be of limited use for positional gene cloning in P. trifoliata and citrus. Analysis of candidate genes for Ctv is also discussed. PMID- 12586874 TI - WVD2 and WDL1 modulate helical organ growth and anisotropic cell expansion in Arabidopsis. AB - Wild-type Arabidopsis roots develop a wavy pattern of growth on tilted agar surfaces. For many Arabidopsis ecotypes, roots also grow askew on such surfaces, typically slanting to the right of the gravity vector. We identified a mutant, wvd2-1, that displays suppressed root waving and leftward root slanting under these conditions. These phenotypes arise from transcriptional activation of the novel WAVE-DAMPENED2 (WVD2) gene by the cauliflower mosaic virus 35S promoter in mutant plants. Seedlings overexpressing WVD2 exhibit constitutive right-handed helical growth in both roots and etiolated hypocotyls, whereas the petioles of WVD2-overexpressing rosette leaves exhibit left-handed twisting. Moreover, the anisotropic expansion of cells is impaired, resulting in the formation of shorter and stockier organs. In roots, the phenotype is accompanied by a change in the arrangement of cortical microtubules within peripheral cap cells and cells at the basal end of the elongation zone. WVD2 transcripts are detectable by reverse transcriptase-polymerase chain reaction in multiple organs of wild-type plants. Its predicted gene product contains a conserved region named "KLEEK," which is found only in plant proteins. The Arabidopsis genome possesses seven other genes predicted to encode KLEEK-containing products. Overexpression of one of these genes, WVD2-LIKE 1, which encodes a protein with regions of similarity to WVD2 extending beyond the KLEEK domain, results in phenotypes that are highly similar to wvd2-1. Silencing of WVD2 and its paralogs results in enhanced root skewing in the wild-type direction. Our observations suggest that at least two members of this gene family may modulate both rotational polarity and anisotropic cell expansion during organ growth. PMID- 12586875 TI - The maize low-phytic acid mutant lpa2 is caused by mutation in an inositol phosphate kinase gene. AB - Reduced phytic acid content in seeds is a desired goal for genetic improvement in several crops. Low-phytic acid mutants have been used in genetic breeding, but it is not known what genes are responsible for the low-phytic acid phenotype. Using a reverse genetics approach, we found that the maize (Zea mays) low-phytic acid lpa2 mutant is caused by mutation in an inositol phosphate kinase gene. The maize inositol phosphate kinase (ZmIpk) gene was identified through sequence comparison with human and Arabidopsis Ins(1,3,4)P(3) 5/6-kinase genes. The purified recombinant ZmIpk protein has kinase activity on several inositol polyphosphates, including Ins(1,3,4)P(3), Ins(3,5,6)P(3), Ins(3,4,5,6)P(4), and Ins(1,2,5,6)P(4). The ZmIpk mRNA is expressed in the embryo, the organ where phytic acid accumulates in maize seeds. The ZmIpk Mutator insertion mutants were identified from a Mutator F(2) family. In the ZmIpk Mu insertion mutants, seed phytic acid content is reduced approximately 30%, and inorganic phosphate is increased about 3-fold. The mutants also accumulate myo-inositol and inositol phosphates as in the lpa2 mutant. Allelic tests showed that the ZmIpk Mu insertion mutants are allelic to the lpa2. Southern-blot analysis, cloning, and sequencing of the ZmIpk gene from lpa2 revealed that the lpa2-1 allele is caused by the genomic sequence rearrangement in the ZmIpk locus and the lpa2-2 allele has a nucleotide mutation that generated a stop codon in the N-terminal region of the ZmIpk open reading frame. These results provide evidence that ZmIpk is one of the kinases responsible for phytic acid biosynthesis in developing maize seeds. PMID- 12586876 TI - Expression of a bifunctional fusion of the Escherichia coli genes for trehalose-6 phosphate synthase and trehalose-6-phosphate phosphatase in transgenic rice plants increases trehalose accumulation and abiotic stress tolerance without stunting growth. AB - Trehalose plays an important role in stress tolerance in plants. Trehalose producing, transgenic rice (Oryza sativa) plants were generated by the introduction of a gene encoding a bifunctional fusion (TPSP) of the trehalose-6 phosphate (T-6-P) synthase (TPS) and T-6-P phosphatase (TPP) of Escherichia coli, under the control of the maize (Zea mays) ubiquitin promoter (Ubi1). The high catalytic efficiency (Seo et al., 2000) of the fusion enzyme and the single-gene engineering strategy make this an attractive candidate for high-level production of trehalose; it has the added advantage of reducing the accumulation of potentially deleterious T-6-P. The trehalose levels in leaf and seed extracts from Ubi1::TPSP plants were increased up to 1.076 mg g fresh weight(-1). This level was 200-fold higher than that of transgenic tobacco (Nicotiana tabacum) plants transformed independently with either TPS or TPP expression cassettes. The carbohydrate profiles were significantly altered in the seeds, but not in the leaves, of Ubi1::TPSP plants. It has been reported that transgenic plants with E. coli TPS and/or TPP were severely stunted and root morphology was altered. Interestingly, our Ubi1::TPSP plants showed no growth inhibition or visible phenotypic alterations despite the high-level production of trehalose. Moreover, trehalose accumulation in Ubi1::TPSP plants resulted in increased tolerance to drought, salt, and cold, as shown by chlorophyll fluorescence and growth inhibition analyses. Thus, our results suggest that trehalose acts as a global protectant against abiotic stress, and that rice is more tolerant to trehalose synthesis than dicots. PMID- 12586877 TI - Characterization of a plant homolog of hop, a cochaperone of hsp90. AB - The 90-kD molecular chaperone hsp90 is the key component of a multiprotein chaperone complex that facilitates folding, stabilization, and functional modulation of a number of signaling proteins. The components of the animal chaperone complex include hsp90, hsp70, hsp40, Hop, and p23. The animal Hop functions to link hsp90 and hsp70, and it can also inhibit the ATPase activity of hsp90. We have demonstrated the presence of an hsp90 chaperone complex in plant cells, but not all components of the complex have been identified. Here, we report the isolation and characterization of soybean (Glycine max) GmHop-1, a soybean homolog of mammalian Hop. An analysis of soybean expressed sequence tags, combined with preexisting data in literature, suggested the presence of at least three related genes encoding Hop-like proteins in soybean. Transcripts corresponding to Hop-like proteins in soybean were detected under normal growth conditions, and their levels increased further in response to stress. A recombinant GmHop-1 bound hsp90 and its binding to hsp90 could be blocked by the tetratricopeptide repeat (TPR) domain of rat (Rattus norvegicus) protein phosphatase 5. Deletion of amino acids 325 to 395, adjacent to the TPR2A domain in GmHop-1, resulted in loss of hsp90 binding. In a minimal assembly system, GmHop-1 was able to stimulate mammalian steroid receptor folding. These data show that plant and animal Hop homologs are conserved in their general characteristics, and suggest that a Hop-like protein in plants is an important cochaperone of plant hsp90. PMID- 12586878 TI - A no hydrotropic response root mutant that responds positively to gravitropism in Arabidopsis. AB - For most plants survival depends upon the capacity of root tips to sense and move towards water and other nutrients in the soil. Because land plants cannot escape environmental stress they use developmental solutions to remodel themselves in order to better adapt to the new conditions. The primary site for perception of underground signals is the root cap (RC). Plant roots have positive hydrotropic response and modify their growth direction in search of water. Using a screening system with a water potential gradient, we isolated a no hydrotropic response (nhr) semi-dominant mutant of Arabidopsis that continued to grow downwardly into the medium with the lowest water potential contrary to the positive hydrotropic and negative gravitropic response seen in wild type-roots. The lack of hydrotropic response of nhr1 roots was confirmed in a system with a gradient in air moisture. The root gravitropic response of nhr1 seedlings was significantly faster in comparison with those of wild type. The frequency of the waving pattern in nhr1 roots was increased compared to those of wild type. nhr1 seedlings had abnormal root cap morphogenesis and reduced root growth sensitivity to abscisic acid (ABA) and the polar auxin transport inhibitor N-(1-naphtyl)phtalamic acid (NPA). These results showed that hydrotropism is amenable to genetic analysis and that an ABA signaling pathway participates in sensing water potential gradients through the root cap. PMID- 12586880 TI - Powdery mildew-induced Mla mRNAs are alternatively spliced and contain multiple upstream open reading frames. AB - In barley (Hordeum vulgare), the Mla13 powdery mildew resistance gene confers Rar1-dependent, AvrMla13-specific resistance to Blumeria graminis f. sp. hordei (Bgh). We have identified cDNA and genomic copies of Mla13 and used this coiled coil nucleotide-binding site leucine-rich repeat protein-encoding gene as a model for the regulation of host resistance to obligate biotrophic fungi in cereals. We demonstrate quantitatively that a rapid increase in the accumulation of Mla transcripts and transcripts of the Mla-signaling genes, Rar1 and Sgt1, is triggered between 16 and 20 h post inoculation, the same time frame that haustoria of avirulent Bgh make contact with the host cell plasma membrane. An abundance of Mla13 cDNAs revealed five classes of transcript leader regions containing two alternatively spliced introns and up to three upstream open reading frames (uORFs). Alternative splicing of introns in the transcript leader region results in a different number of uORFs and variability in the size of uORF2. These results indicate that regulation of Mla transcript accumulation is not constitutive and that induction is coordinately controlled by recognition specific factors. The sudden increase in specific transcript levels could account for the rapid defense response phenotype conferred by Mla6 and Mla13. PMID- 12586879 TI - AtCSLA7, a cellulose synthase-like putative glycosyltransferase, is important for pollen tube growth and embryogenesis in Arabidopsis. AB - The cellulose synthase-like proteins are a large family of proteins in plants thought to be processive polysaccharide beta-glycosyltransferases. We have characterized an Arabidopsis mutant with a transposon insertion in the gene encoding AtCSLA7 of the CSLA subfamily. Analysis of the transmission efficiency of the insertion indicated that AtCSLA7 is important for pollen tube growth. Moreover, the homozygous insertion was embryo lethal. A detailed analysis of seed developmental progression revealed that mutant embryos developed more slowly than wild-type siblings. The mutant embryos also showed abnormal cell patterning and they arrested at a globular stage. The defective embryonic development was associated with reduced proliferation and failed cellularization of the endosperm. AtCSLA7 is widely expressed, and is likely to be required for synthesis of a cell wall polysaccharide found throughout the plant. Our results suggest that this polysaccharide is essential for cell wall structure or for signaling during plant embryo development. PMID- 12586882 TI - Differences in whole-cell and single-channel ion currents across the plasma membrane of mesophyll cells from two closely related Thlaspi species. AB - The patch clamp technique was used to study the physiology of ion transport in mesophyll cells from two Thlaspi spp. that differ significantly in their physiology. In comparison with Thlaspi arvense, Thlaspi caerulescens (a heavy metal accumulator) can grow in, tolerate, and accumulate very high levels of certain heavy metals (primarily zinc [Zn] and cadmium) in their leaf cells. The membrane conductance of every T. arvense leaf cell was dominated by a slowly activating, time-dependent outward rectifying current (SKOR). In contrast, only 23% of T. caerulescens cells showed SKOR activity, whereas the remaining 77% exhibit a rapidly developing instantaneous K(+) outward rectifier (RKOR) current. In contrast to RKOR, the channels underlying the SKOR current were sensitive to changes in the extracellular ion activity. Single-channel recordings indicated the existence of K(+) channel populations with similar unitary conductances, but distinct channel kinetics and regulation. The correlation between these recordings and the whole-cell data indicated that although one type of channel kinetics is preferentially activated in each Thlaspi spp., both species have the capability to switch between either type of current. Ion substitution in whole cell and single-channel experiments indicated that although the SKOR and RKOR channels mediate a net outward K(+) current, they can also allow a significant Zn(2+) permeation (i.e. influx). In addition, single-channel recordings allowed us to identify an infrequent type of plasma membrane divalent cation channel that also can mediate Zn(2+) influx. We propose that the different K(+) channel types or channel states may result from and are likely to reflect differences in the cytoplasmic and apoplastic ionic environment in each species. Thus, the ability to interchangeably switch between different channel states allows each species to constantly adjust to changes in their apoplastic ionic environment. PMID- 12586881 TI - Comparative transcriptional profiling of placenta and endosperm in developing maize kernels in response to water deficit. AB - The early post-pollination phase of maize (Zea mays) development is particularly sensitive to water deficit stress. Using cDNA microarray, we studied transcriptional profiles of endosperm and placenta/pedicel tissues in developing maize kernels under water stress. At 9 d after pollination (DAP), placenta/pedicel and endosperm differed considerably in their transcriptional responses. In placenta/pedicel, 79 genes were significantly affected by stress and of these 89% were up-regulated, whereas in endosperm, 56 genes were significantly affected and 82% of these were down-regulated. Only nine of the stress-regulated genes were in common between these tissues. Hierarchical cluster analysis indicated that different sets of genes were regulated in the two tissues. After rewatering at 9 DAP, profiles at 12 DAP suggested that two regulons exist, one for genes responding specifically to concurrent imposition of stress, and another for genes remaining affected after transient stress. In placenta, genes encoding recognized stress tolerance proteins, including heat shock proteins, chaperonins, and major intrinsic proteins, were the largest class of genes regulated, all of which were up-regulated. In contrast, in endosperm, genes in the cell division and growth category represented a large class of down regulated genes. Several cell wall-degrading enzymes were expressed at lower levels than in controls, suggesting that stress delayed normal advance to programmed cell death in the central endosperm. We suggest that the responsiveness of placenta to whole-plant stress factors (water potential, abscisic acid, and sugar flux) and of endosperm to indirect factors may play key roles in determining the threshold for kernel abortion. PMID- 12586883 TI - Zinc efficiency is correlated with enhanced expression and activity of zinc requiring enzymes in wheat. AB - Zinc (Zn) is an essential micronutrient for plants. The ability of plants to maintain significant yields under low Zn is termed Zn efficiency (ZE) and its genetic and mechanistic basis is still not well understood. Previously, we showed that root Zn uptake did not play a role in ZE. In the current study, Zn-efficient and -inefficient wheat (Triticum aestivum) genotypes were grown for 13 d in chelate buffer nutrient solutions at low (0.1 pM), sufficient (150 pM), and high (1 microM) Zn(2+) activities and analyzed for root-to-shoot translocation of Zn, subcellular leaf Zn distribution, and activity and expression of the Zn-requiring enzymes in leaves. No correlation between ZE and Zn translocation to the shoot was found. Furthermore, total and water-soluble concentrations of leaf Zn were not associated with ZE, and no differences in subcellular Zn compartmentation were found between Zn-efficient and -inefficient genotypes. However, the expression and activity of the Zn-requiring enzymes copper (Cu)/Zn superoxide dismutase (SOD) and carbonic anhydrase did correlate with differences in ZE. Northern analysis suggested that Cu/ZnSOD gene expression was up-regulated in the Zn-efficient genotype, Kirgiz, but not in inefficient BDME. Under Zn deficiency stress, the very Zn-efficient genotype Kirgiz and moderately Zn-efficient Dagdas exhibited an increased activity of Cu/ZnSOD and carbonic anhydrase when compared with Zn-inefficient BDME. These results suggest that Zn-efficient genotypes may be able to maintain the functioning of Zn-requiring enzymes under low Zn conditions; thus, biochemical Zn utilization may be an important component of ZE in wheat. PMID- 12586884 TI - Functional divergence of a syntenic invertase gene family in tomato, potato, and Arabidopsis. AB - Comparative analysis of complex developmental pathways depends on our ability to resolve the function of members of gene families across taxonomic groups. LIN5, which belongs to a small gene family of apoplastic invertases in tomato (Lycopersicon esculentum), is a quantitative trait locus that modifies fruit sugar composition. We have compared the genomic organization and expression of this gene family in the two distantly related species: tomato and Arabidopsis. Invertase family members reside on segmental duplications in the near-colinear genomes of tomato and potato (Solanum tuberosum). These chromosomal segments are syntenically duplicated in the model plant Arabidopsis. On the basis of phylogenetic analysis of genes in the microsyntenic region, we conclude that these segmental duplications arose independently after the separation of the tomato/potato clade from Arabidopsis. Rapid regulatory divergence is characteristic of the invertase family. Interestingly, although the processes of gene duplication and specialization of expression occurred separately in the two species, synteny-based orthologs from both clades acquired similar organ-specific expression. This similar expression pattern of the genes is evidence of comparable evolutionary constraints (parallel evolution) rather than of functional orthology. The observation that functional orthology cannot be identified through analysis of expression similarity highlights the caution that needs to be exercised in extrapolating developmental networks from a model organism. PMID- 12586885 TI - Recently duplicated maize R2R3 Myb genes provide evidence for distinct mechanisms of evolutionary divergence after duplication. AB - R2R3 Myb genes are widely distributed in the higher plants and comprise one of the largest known families of regulatory proteins. Here, we provide an evolutionary framework that helps explain the origin of the plant-specific R2R3 Myb genes from widely distributed R1R2R3 Myb genes, through a series of well established steps. To understand the routes of sequence divergence that followed Myb gene duplication, we supplemented the information available on recently duplicated maize (Zea mays) R2R3 Myb genes (C1/Pl1 and P1/P2) by cloning and characterizing ZmMyb-IF35 and ZmMyb-IF25. These two genes correspond to the recently expanded P-to-A group of maize R2R3 Myb genes. Although the origins of C1/Pl1 and ZmMyb-IF35/ZmMyb-IF25 are associated with the segmental allotetraploid origin of the maize genome, other gene duplication events also shaped the P-to-A clade. Our analyses indicate that some recently duplicated Myb gene pairs display substantial differences in the numbers of synonymous substitutions that have accumulated in the conserved MYB domain and the divergent C-terminal regions. Thus, differences in the accumulation of substitutions during evolution can explain in part the rapid divergence of C-terminal regions for these proteins in some cases. Contrary to previous studies, we show that the divergent C termini of these R2R3 MYB proteins are subject to purifying selection. Our results provide an in-depth analysis of the sequence divergence for some recently duplicated R2R3 Myb genes, yielding important information on general patterns of evolution for this large family of plant regulatory genes. PMID- 12586886 TI - Fructan 1-exohydrolases. beta-(2,1)-trimmers during graminan biosynthesis in stems of wheat? Purification, characterization, mass mapping, and cloning of two fructan 1-exohydrolase isoforms. AB - Graminan-type fructans are temporarily stored in wheat (Triticum aestivum) stems. Two phases can be distinguished: a phase of fructan biosynthesis (green stems) followed by a breakdown phase (stems turning yellow). So far, no plant fructan exohydrolase enzymes have been cloned from a monocotyledonous species. Here, we report on the cloning, purification, and characterization of two fructan 1 exohydrolase cDNAs (1-FEH w1 and w2) from winter wheat stems. Similar to dicot plant 1-FEHs, they are derived from a special group within the cell wall-type invertases characterized by their low isoelectric points. The corresponding isoenzymes were purified to electrophoretic homogeneity, and their mass spectra were determined by quadrupole-time-of-flight mass spectrometry. Characterization of the purified enzymes revealed that inulin-type fructans [beta-(2,1)] are much better substrates than levan-type fructans [beta-(2,6)]. Although both enzymes are highly identical (98% identity), they showed different substrate specificity toward branched wheat stem fructans. Although 1-FEH activities were found to be considerably higher during the fructan breakdown phase, it was possible to purify substantial amounts of 1-FEH w2 from young, fructan biosynthesizing wheat stems, suggesting that this isoenzyme might play a role as a beta-(2,1)-trimmer throughout the period of active graminan biosynthesis. In this way, the species and developmental stage-specific complex fructan patterns found in monocots might be determined by the relative proportions and specificities of both fructan biosynthetic and breakdown enzymes. PMID- 12586887 TI - Homogentisate phytyltransferase activity is limiting for tocopherol biosynthesis in Arabidopsis. AB - Tocopherols are essential components of the human diet and are synthesized exclusively by photosynthetic organisms. These lipophilic antioxidants consist of a chromanol ring and a 15-carbon tail derived from homogentisate (HGA) and phytyl diphosphate, respectively. Condensation of HGA and phytyl diphosphate, the committed step in tocopherol biosynthesis, is catalyzed by HGA phytyltransferase (HPT). To investigate whether HPT activity is limiting for tocopherol synthesis in plants, the gene encoding Arabidopsis HPT, HPT1, was constitutively overexpressed in Arabidopsis. In leaves, HPT1 overexpression resulted in a 10 fold increase in HPT specific activity and a 4.4-fold increase in total tocopherol content relative to wild type. In seeds, HPT1 overexpression resulted in a 4-fold increase in HPT specific activity and a total seed tocopherol content that was 40% higher than wild type, primarily because of an increase in gamma tocopherol content. This enlarged pool of gamma-tocopherol was almost entirely converted to alpha-tocopherol by crossing HPT1 overexpressing plants with lines constitutively overexpressing gamma-tocopherol methyltransferase. Seed of the resulting double overexpressing lines had a 12-fold increase in vitamin E activity relative to wild type. These results indicate that HPT activity is limiting in various Arabidopsis tissues and that total tocopherol levels and vitamin E activity can be elevated in leaves and seeds by combined overexpression of the HPT1 and gamma-tocopherol methyltransferase genes. PMID- 12586888 TI - The Arabidopsis STICHEL gene is a regulator of trichome branch number and encodes a novel protein. AB - Here, we analyze the STICHEL (STI) gene, which plays an important role in the regulation of branch number of the unicellular trichomes in Arabidopsis. We have isolated the STI locus by positional cloning and confirmed the identity by sequencing seven independent sti alleles. The STI gene encodes a protein of 1,218 amino acid residues containing a domain with sequence similarity to the ATP binding eubacterial DNA-polymerase III gamma-subunits. Because endoreduplication was found to be normal in sti mutants the molecular function of STI in cell morphogenesis is not linked to DNA replication and, therefore, postulated to represent a novel pathway. Northern-blot analysis shows that STI is expressed in all organs suggesting that STI function is not trichome specific. The analysis of sti alleles and transgenic lines overexpressing STI suggests that STI regulates branching in a dosage-dependent manner. PMID- 12586889 TI - Overexpression of Arabidopsis phytochelatin synthase paradoxically leads to hypersensitivity to cadmium stress. AB - Phytochelatin (PC) plays an important role in heavy metal detoxification in plants and other living organisms. Therefore, we overexpressed an Arabidopsis PC synthase (AtPCS1) in transgenic Arabidopsis with the goal of increasing PC synthesis, metal accumulation, and metal tolerance in these plants. Transgenic Arabidopsis plants were selected, designated pcs lines, and analyzed for tolerance to cadmium (Cd). Transgenic pcs lines showed 12- to 25-fold higher accumulation of AtPCS1 mRNA, and production of PCs increased by 1.3- to 2.1-fold under 85 microM CdCl(2) stress for 3 d when compared with wild-type plants. Cd tolerance was assessed by measuring root length of plants grown on agar medium containing 50 or 85 microM CdCl(2). Pcs lines paradoxically showed hypersensitivity to Cd stress. This hypersensitivity was also observed for zinc (Zn) but not for copper (Cu). The overexpressed AtPCS1 protein itself was not responsible for Cd hypersensitivity as transgenic cad1-3 mutants overexpressing AtPCS1 to similar levels as those of pcs lines were not hypersensitive to Cd. Pcs lines were more sensitive to Cd than a PC-deficient Arabidopsis mutant, cad1-3, grown under low glutathione (GSH) levels. Cd hypersensitivity of pcs lines disappeared under increased GSH levels supplemented in the medium. Therefore, Cd hypersensitivity in pcs lines seems due to the toxicity of PCs as they existed at supraoptimal levels when compared with GSH levels. PMID- 12586890 TI - Combining quantitative trait Loci analysis and an ecophysiological model to analyze the genetic variability of the responses of maize leaf growth to temperature and water deficit. AB - Ecophysiological models predict quantitative traits of one genotype in any environment, whereas quantitative trait locus (QTL) models predict the contribution of alleles to quantitative traits under a limited number of environments. We have combined both approaches by dissecting into effects of QTLs the parameters of a model of maize (Zea mays) leaf elongation rate (LER; H. Ben Haj Salah, F. Tardieu [1997] Plant Physiol 114: 893-900). Response curves of LER to meristem temperature, water vapor pressure difference, and soil water status were established in 100 recombinant inbred lines (RILs) of maize in six experiments carried out in the field or in the greenhouse. All responses were linear and common to different experiments, consistent with the model. A QTL analysis was carried out on the slopes of these responses by composite interval mapping confirmed by bootstrap analysis. Most QTLs were specific of one response only. QTLs of abscisic acid concentration in the xylem sap colocalized with QTLs of response to soil water deficit and conferred a low response. Each parameter of the ecophysiological model was computed as the sum of QTL effects, allowing calculation of parameters for 11 new RILs and two parental lines. LERs were simulated and compared with measurements in a growth chamber experiment. The combined model accounted for 74% of the variability of LER, suggesting that it has a general value for any RIL under any environment. PMID- 12586891 TI - Single-cell measurements of the contributions of cytosolic Na(+) and K(+) to salt tolerance. AB - Ion concentrations in the roots of two barley (Hordeum vulgare) varieties that differed in NaCl tolerance were compared after exposure to NaCl. Triple-barreled H(+)-, K(+)-, and Na(+)-selective microelectrodes were used to measure cytosolic activities of the three ions after 5 and 8 d of NaCl stress. In both varieties of barley, it was only possible to record successfully from root cortical cells because the epidermal cells appeared to be damaged. The data show that from the 1st d of full NaCl stress, there were differences in the way in which the two varieties responded. At 5 d, the tolerant variety maintained a 10-fold lower cytosolic Na(+) than the more sensitive variety, although by 8 d the two varieties were not significantly different. At this time, the more tolerant variety was better at maintaining root cytosolic K(+) in the high-NaCl background than was the more sensitive variety. In contrast to earlier work on K(+)-starved barley (Walker et al., 1996), there was no acidification of the cytosol associated with the decreased cytosolic K(+) activity during NaCl stress. These single-cell measurements of cytosolic and vacuolar ion activities allow calculation of thermodynamic gradients that can be used to reveal (or predict) the type of active transporters at both the plasma membrane and tonoplast. PMID- 12586892 TI - A low-starch barley mutant, riso 16, lacking the cytosolic small subunit of ADP glucose pyrophosphorylase, reveals the importance of the cytosolic isoform and the identity of the plastidial small subunit. AB - To provide information on the roles of the different forms of ADP-glucose pyrophosphorylase (AGPase) in barley (Hordeum vulgare) endosperm and the nature of the genes encoding their subunits, a mutant of barley, Riso 16, lacking cytosolic AGPase activity in the endosperm was identified. The mutation specifically abolishes the small subunit of the cytosolic AGPase and is attributable to a large deletion within the coding region of a previously characterized small subunit gene that we have called Hv.AGP.S.1. The plastidial AGPase activity in the mutant is unaffected. This shows that the cytosolic and plastidial small subunits of AGPase are encoded by separate genes. We purified the plastidial AGPase protein and, using amino acid sequence information, we identified the novel small subunit gene that encodes this protein. Studies of the Riso 16 mutant revealed the following. First, the reduced starch content of the mutant showed that a cytosolic AGPase is required to achieve the normal rate of starch synthesis. Second, the mutant makes both A- and B-type starch granules, showing that the cytosolic AGPase is not necessary for the synthesis of these two granule types. Third, analysis of the phylogenetic relationships between the various small subunit proteins both within and between species, suggest that the cytosolic AGPase single small subunit gene probably evolved from a leaf single small subunit gene. PMID- 12586893 TI - Zonal changes in ascorbate and hydrogen peroxide contents, peroxidase, and ascorbate-related enzyme activities in onion roots. AB - Onion (Allium cepa) roots growing hydroponically show differential zonal values for intra- (symplastic) and extra- (apoplastic) cellular ascorbate (ASC) and dehydroascorbate (DHA) contents and for related enzyme activities. In whole roots, ASC and DHA concentrations were higher in root apex and meristem and gradually decreased toward the root base. Guaiacol peroxidase, ASC peroxidase, monodehydroascorbate oxidoreductase, DHA reductase, catalase, and glutathione reductase activities showed differential activity patterns depending on the zone of the root and their apoplastic or symplastic origin. An in vivo staining of peroxidase activity also revealed a specific distribution pattern along the root axis. Using electron microscopy, hydrogen peroxide was found at different locations depending on the root zone but was mainly located in cell walls from epidermal and meristematic cells and in cells undergoing lignification. A balanced control of all of these molecules seems to exist along the root axis and may be directly related to the mechanisms in which the ASC system is involved, as cell division and elongation. The role of ASC on growth and development in relation to its presence at the different zones of the root is discussed. PMID- 12586894 TI - Arabidopsis ICX1 is a negative regulator of several pathways regulating flavonoid biosynthesis genes. AB - Flavonoid biosynthesis gene expression is controlled by a range of endogenous and environmental signals. The Arabidopsis icx1 (increased chalcone synthase expression 1) mutant has elevated induction of CHS (CHALCONE SYNTHASE) and other flavonoid biosynthesis genes in response to several stimuli. We show that ICX1 is a negative regulator of the cryptochrome 1, phytochrome A, ultraviolet (UV)-B, low temperature, sucrose, and cytokinin induction of CHS expression and/or anthocyanin accumulation, demonstrating that these pathways are regulated either directly or indirectly by at least one common component. Expression analysis of CHS and other genes (LTP, CAB, and rbcS) indicates that ICX1 functions in both seedlings and mature leaf tissue and acts principally in the epidermis, consistent with the alterations in epidermal development seen in icx1. The mutant was unaltered in the synergistic interactions between UV-B, blue, and UV-A light that regulate CHS and we propose a model of action of ICX1 in these responses. PMID- 12586895 TI - The role of free histidine in xylem loading of nickel in Alyssum lesbiacum and Brassica juncea. AB - Exposure of the hyperaccumulator Alyssum lesbiacum to nickel (Ni) is known to result in a dose-dependent increase in xylem sap concentrations of Ni and the chelator free histidine (His). Addition of equimolar concentrations of exogenous L-His to an Ni-amended hydroponic rooting medium enhances Ni flux into the xylem in the nonaccumulator Alyssum montanum, and, as reported here, in Brassica juncea L. cv Vitasso. In B. juncea, reducing the entry of L-His into the root by supplying D-His instead of L-His, or L-His in the presence of a 10-fold excess of L-alanine, did not affect root Ni uptake, but reduced Ni release into the xylem. Compared with B. juncea, root His concentrations were constitutively about 4.4 fold higher in A. lesbiacum, and did not increase within 9 h of exposure to Ni. Cycloheximide did not affect root His or Ni concentrations, but strongly decreased the release of His and Ni from the root into the xylem of A. lesbiacum, whereas xylem sap concentrations of Ca and Mg remained unaffected. Near quantitative chelation of Ni with nitrilotriacetate in the rooting medium did not enhance Ni flux into the xylem of A. lesbiacum and B. juncea, suggesting the absence of a significant apoplastic pathway for Ni entry into the xylem. The data suggest that in B. juncea roots, Ni(2+) uptake is independent of simultaneous uptake of His. In both species, enhanced release of Ni into the xylem is associated with concurrent release of His from an increased root free His pool. PMID- 12586897 TI - Xylem ray parenchyma cells in boreal hardwood species respond to subfreezing temperatures by deep supercooling that is accompanied by incomplete desiccation. AB - It has been accepted that xylem ray parenchyma cells (XRPCs) in hardwood species respond to subfreezing temperatures either by deep supercooling or by extracellular freezing. Present study by cryo-scanning electron microscopy examined the freezing responses of XRPCs in five boreal hardwoods: Salix sachalinensis Fr. Schmit, Populus sieboldii Miq., Betula platyphylla Sukat. var japonica Hara, Betula pubescens Ehrh., and red osier dogwood (Cornus sericea), in which XRPCs have been reported to respond by extracellular freezing. Cryo scanning electron microscopy observations revealed that slow cooling of xylem to 80 degrees C resulted in intracellular freezing in the majority of XRPCs in S. sachalinensis, an indication that these XRPCs had been deep supercooled. In contrast, in the majority of XRPCs in P. sieboldii, B. platyphylla, B. pubescens, and red osier dogwood, slow cooling to -80 degrees C produced slight cytorrhysis without clear evidence of intracellular freezing, suggesting that these XRPCs might respond by extracellular freezing. In these XRPCs exhibited putative extracellular freezing; however, deep etching revealed the apparent formation of intracellular ice crystals in restricted local areas. To confirm the occurrence of intracellular freezing, we rewarmed these XRPCs after cooling and observed very large intracellular ice crystals as a result of the recrystallization. Thus, the XRPCs in all the boreal hardwoods that we examined responded by deep supercooling that was accompanied with incomplete desiccation. From these results, it seems possible that limitations to the deep-supercooling ability of XRPCs might be a limiting factor for adaptation of hardwoods to cold climates. PMID- 12586896 TI - A novel small heat shock protein gene, vis1, contributes to pectin depolymerization and juice viscosity in tomato fruit. AB - We have characterized a novel small heat shock protein gene, viscosity 1 (vis1) from tomato (Lycopersicon esculentum) and provide evidence that it plays a role in pectin depolymerization and juice viscosity in ripening fruits. Expression of vis1 is negatively associated with juice viscosity in diverse tomato genotypes. vis1 exhibits DNA polymorphism among tomato genotypes, and the alleles vis1-hta (high-transcript accumulator; accession no. AY128101) and vis1-lta (low transcript accumulator; accession no. AY128102) are associated with thinner and thicker juice, respectively. Segregation of tomato lines heterogeneous for vis1 alleles indicates that vis1 influences pectin depolymerization and juice viscosity in ripening fruits. vis1 is regulated by fruit ripening and high temperature and exhibits a typical heat shock protein chaperone function when expressed in bacterial cells. We propose that VIS1 contributes to physiochemical properties of juice, including pectin depolymerization, by reducing thermal denaturation of depolymerizing enzymes during daytime elevated temperatures. PMID- 12586898 TI - Sudden exposure to solar UV-B radiation reduces net CO(2) uptake and photosystem I efficiency in shade-acclimated tropical tree seedlings. AB - Tree seedlings developing in the understory of the tropical forest have to endure short periods of high-light stress when tree-fall gaps are formed, and direct solar radiation, including substantial UV light, reaches the leaves. In experiments simulating the opening of a tree-fall gap, the response of photosynthesis in leaves of shade-acclimated seedlings (Anacardium excelsum, Virola surinamensis, and Calophyllum longifolium) to exposure to direct sunlight (for 20-50 min) was investigated in Panama (9 degrees N). To assess the effects of solar UV-B radiation (280-320 nm), the sunlight was filtered through plastic films that selectively absorbed UV-B or transmitted the complete spectrum. The results document a strong inhibition of CO(2) assimilation by sun exposure. Light limited and light-saturated rates of photosynthetic CO(2) uptake by the leaves were affected, which apparently occurred independently of a simultaneous inhibition of potential photosystem (PS) II efficiency. The ambient UV-B light substantially contributed to these effects. The photochemical capacity of PSI, measured as absorbance change at 810 nm in saturating far-red light, was not significantly affected by sun exposure of the seedlings. However, a decrease in the efficiency of P700 photooxidation by far-red light was observed, which was strongly promoted by solar UV-B radiation. The decrease in PSI efficiency may result from enhanced charge recombination in the reaction center, which might represent an incipient inactivation of PSI, but contributes to thermal dissipation of excessive light energy and thereby to photoprotection. PMID- 12586899 TI - A new type of a multifunctional beta-oxidation enzyme in euglena. AB - The biochemical and molecular properties of the beta-oxidation enzymes from algae have not been investigated yet. The present study provides such data for the phylogenetically old alga Euglena (Euglena gracilis). A novel multifunctional beta-oxidation complex was purified to homogeneity by ammonium sulfate precipitation, density gradient centrifugation, and ion-exchange chromatography. Monospecific antibodies used in immunocytochemical experiments revealed that the enzyme is located in mitochondria. The enzyme complex is composed of 3 hydroxyacyl-coenzyme A (-CoA) dehydrogenase, 2-enoyl-CoA hydratase, thiolase, and epimerase activities. The purified enzyme exhibits a native molecular mass of about 460 kD, consisting of 45.5-, 44.5-, 34-, and 32-kD subunits. Subunits dissociated from the complete complex revealed that the hydratase and the thiolase functions are located on the large subunits, whereas two dehydrogenase functions are located on the two smaller subunits. Epimerase activity was only measurable in the complete enzyme complex. From the use of stereoisomers and sequence data, it was concluded that the 2-enoyl-CoA hydratase catalyzes the formation of L-hydroxyacyl CoA isomers and that both of the different 3 hydroxyacyl-CoA dehydrogenase functions on the 32- and 34-kD subunits are specific to L-isomers as substrates, respectively. All of these data suggest that the Euglena enzyme belongs to the family of beta-oxidation enzymes that degrade acyl-CoAs via L-isomers and that it is composed of subunits comparable with subunits of monofunctional beta-oxidation enzymes. It is concluded that the Euglena enzyme phylogenetically developed from monospecific enzymes in archeons by non-covalent combination of subunits and presents an additional line for the evolutionary development of multifunctional beta-oxidation enzymes. PMID- 12586901 TI - Metabolic engineering of valine- and isoleucine-derived glucosinolates in Arabidopsis expressing CYP79D2 from Cassava. AB - Glucosinolates are amino acid-derived natural products that, upon hydrolysis, typically release isothiocyanates with a wide range of biological activities. Glucosinolates play a role in plant defense as attractants and deterrents against herbivores and pathogens. A key step in glucosinolate biosynthesis is the conversion of amino acids to the corresponding aldoximes, which is catalyzed by cytochromes P450 belonging to the CYP79 family. Expression of CYP79D2 from cassava (Manihot esculenta Crantz.) in Arabidopsis resulted in the production of valine (Val)- and isoleucine-derived glucosinolates not normally found in this ecotype. The transgenic lines showed no morphological phenotype, and the level of endogenous glucosinolates was not affected. The novel glucosinolates were shown to constitute up to 35% of the total glucosinolate content in mature rosette leaves and up to 48% in old leaves. Furthermore, at increased concentrations of these glucosinolates, the proportion of Val-derived glucosinolates decreased. As the isothiocyanates produced from the Val- and isoleucine-derived glucosinolates are volatile, metabolically engineered plants producing these glucosinolates have acquired novel properties with great potential for improvement of resistance to herbivorous insects and for biofumigation. PMID- 12586900 TI - Expression of cytosolic and plastid acetyl-coenzyme A carboxylase genes in young wheat plants. AB - Expression of cytosolic and plastid acetyl-coenzyme A carboxylase (ACCase) gene families at the mRNA level was analyzed in developing wheat (Triticum aestivum) plants. The major plastid ACCase mRNA level is high in the middle part of the plant and low in roots and leaf blades. An alternative plastid ACCase transcript initiated at a different promoter and using an alternative 5' splice site for the first intron accumulates to its highest level in roots. Cytosolic ACCase mRNA also consists of two species, one of which is present at approximately a constant level, whereas the other accumulates to a high level in the lower sheath section. It is likely that different promoters are also responsible for the two forms of cytosolic ACCase mRNA. The abundances of cytosolic and plastid ACCase mRNAs in the sheath section of the plant are similar. ACCase protein level is significantly lower in the leaf blades, in parallel with changes in the total ACCase mRNA level. Homoeologous ACCase genes show the same expression patterns and similar mRNA levels, suggesting that none of the genes was silenced or acquired new tissue specificity after polyploidization. PMID- 12586902 TI - Winter at the alpine timberline. Why does embolism occur in norway spruce but not in stone pine? AB - Conifers growing at the alpine timberline are exposed to frost drought and freeze thaw cycles during winter-stress factors known to induce embolism in tree xylem. The two dominant species of the European Central Alps timberline were studied: Norway spruce (Picea abies [L.] Karst) and stone pine (Pinus cembra), which usually reaches higher altitudes. We hypothesized to find embolism only at the timberline and to observe less embolism in stone pine than in Norway spruce due to avoidance mechanisms. Seasonal courses of embolism and water potential were studied at 1,700 and 2,100 m during two winter seasons and correlated to vulnerability (to drought-induced embolism), leaf conductance, and micrometeorological data. Embolism was observed only at the timberline and only in Norway spruce (up to 49.2% loss of conductivity). Conductivity losses corresponded to low water potentials (down to -3.5 MPa) but also to the number of freeze-thaw events indicating both stress factors to contribute to embolism induction. Decreasing embolism rates-probably due to refilling-were observed already in winter. Stone pine did not exhibit an adapted vulnerability (50% loss of conductivity at -3.5 MPa) but avoided critical potentials (minimum -2.3 MPa): Cuticulare conductance was 3.5-fold lower than in Norway spruce, and angles between needles and axes were found to decrease in dehydrating branches. The extent of conductivity losses in Norway spruce and the spectrum of avoidance and recovery mechanisms in both species indicates winter embolism to be relevant for tree line formation. PMID- 12586903 TI - cor Gene expression in barley mutants affected in chloroplast development and photosynthetic electron transport. AB - The expression of several barley (Hordeum vulgare) cold-regulated (cor) genes during cold acclimation was blocked in the albino mutant a(n), implying a chloroplast control on mRNAs accumulation. By using albino and xantha mutants ordered according to the step in chloroplast biogenesis affected, we show that the cold-dependent accumulation of cor14b, tmc-ap3, and blt14 mRNAs depends on plastid developmental stage. Plants acquire the ability to fully express cor genes only after the development of primary thylakoid membranes in their chloroplasts. To investigate the chloroplast-dependent mechanism involved in cor gene expression, the activity of a 643-bp cor14b promoter fragment was assayed in wild-type and albino mutant a(n) leaf explants using transient beta-glucuronidase reporter expression assay. Deletion analysis identified a 27-bp region between nucleotides -274 and -247 with respect to the transcription start point, encompassing a boundary of some element that contributes to the cold-induced expression of cor14b. However, cor14b promoter was equally active in green and in albino a(n) leaves, suggesting that chloroplast controls cor14b expression by posttranscriptional mechanisms. Barley mutants lacking either photosystem I or II reaction center complexes were then used to evaluate the effects of redox state of electron transport chain components on COR14b accumulation. In the mutants analyzed, the amount of COR14b protein, but not the steady-state level of the corresponding mRNA, was dependent on the redox state of the electron transport chain. Treatments of the vir-zb63 mutant with electron transport chain inhibitors showed that oxidized plastoquinone promotes COR14b accumulation, thus suggesting a molecular relationship between plastoquinone/plastoquinol pool and COR14b. PMID- 12586904 TI - Polyamine oxidase, a hydrogen peroxide-producing enzyme, is up-regulated by light and down-regulated by auxin in the outer tissues of the maize mesocotyl. AB - Exogenously supplied auxin (1-naphthaleneacetic acid) inhibited light-induced activity increase of polyamine oxidase (PAO), a hydrogen peroxide-producing enzyme, in the outer tissues of maize (Zea mays) mesocotyl. The same phenomenon operates at PAO protein and mRNA accumulation levels. The wall-bound to extractable PAO activity ratio was unaffected by auxin treatment, either in the dark or after light exposure. Ethylene treatment did not affect PAO activity, thus excluding an effect of auxin via increased ethylene biosynthesis. The auxin polar transport inhibitors N(1)-naphthylphthalamic acid or 2,3,5-triiodobenzoic acid caused a further increase of PAO expression in outer tissues after light treatment. The small increase of PAO expression, normally occurring in the mesocotyl epidermis during plant development in the dark, was also inhibited by auxin, although to a lesser extent with respect to light-exposed tissue, and was stimulated by N(1)-naphthylphthalamic acid or 2,3,5-triiodobenzoic acid, thus suggesting a complex regulation of PAO expression. Immunogold ultrastructural analysis in epidermal cells revealed the association of PAO with the secretory pathway and the cell walls. The presence of the enzyme in the cell walls of this tissue greatly increased in response to light treatment. Consistent with auxin effects on light-induced PAO expression, the hormone treatment inhibited the increase in immunogold staining both intraprotoplasmically and in the cell wall. These results suggest that both light and auxin finely tune PAO expression during the light-induced differentiation of the cell wall in the maize mesocotyl epidermal tissues. PMID- 12586905 TI - Rapid alkalinization factors in poplar cell cultures. Peptide isolation, cDNA cloning, and differential expression in leaves and methyl jasmonate-treated cells. AB - A family of peptides inducing rapid pH alkalinization in hybrid poplar (Populus trichocarpa x Populus deltoides) cell culture medium was isolated from hybrid poplar leaves. Five related approximately 5-kD peptides were purified by high performance liquid chromatography and analyzed by matrix-assisted laser desorption ionization-mass spectrometry. The N-terminal sequence of one of the isolated peptides was very similar to a previously characterized peptide from tobacco (Nicotiana tabacum), rapid alkalinization factor (RALF), which causes a rapid increase in culture medium pH when added to tobacco cell cultures (G. Pearce, D.S. Moura, J. Stratmann, C.A. Ryan [2001] Proc Natl Acad Sci USA 98: 12843-12847). Two unique poplar RALF cDNAs (PtdRALF1 and PtdRALF2) were isolated from a poplar cDNA library and used to study RALF expression in poplar saplings and cultured poplar cells. Both genes were found to be expressed constitutively in poplar saplings and cultured cells. However, PtdRALF2 was expressed in leaves at very low levels, and its expression in suspension culture cells was transiently suppressed by methyl jasmonate (MeJa). Although the function of these novel peptides remains enigmatic, our experiments suggest their role may be developmental rather than stress related. Overall, our study confirms the presence of active RALF peptides in other plants, and provides new data on the complexity of the RALF gene family in poplar. PMID- 12586907 TI - Public engagement with science. PMID- 12586906 TI - In plants, 3-o-methylglucose is phosphorylated by hexokinase but not perceived as a sugar. AB - In plants, sugars are the main respiratory substrates and important signaling molecules in the regulation of carbon metabolism. Sugar signaling studies suggested that sugar sensing involves several key components, among them hexokinase (HXK). Although the sensing mechanism of HXK is unknown, several experiments support the hypothesis that hexose phosphorylation is a determining factor. Glucose (Glc) analogs transported into cells but not phosphorylated are frequently used to test this hypothesis, among them 3-O-methyl-Glc (3-OMG). The aim of the present work was to investigate the effects and fate of 3-OMG in heterotrophic plant cells. Measurements of respiration rates, protein and metabolite contents, and protease activities and amounts showed that 3-OMG is not a respiratory substrate and does not contribute to biosynthesis. Proteolysis and lipolysis are induced in 3-OMG-fed maize (Zea mays L. cv DEA) roots in the same way as in sugar-starved organs. However, contrary to the generally accepted idea, phosphorous and carbon nuclear magnetic resonance experiments and enzymatic assays prove that 3-OMG is phosphorylated to 3-OMG-6-phosphate, which accumulates in the cells. Insofar as plant HXK is involved in sugar sensing, these findings are discussed on the basis of the kinetic properties because the catalytic efficiency of HXK isolated from maize root tips is five orders of magnitude lower for 3-OMG than for Glc and Man. PMID- 12586908 TI - Science publishing. The UPSIDE of good behavior: make your data freely available. PMID- 12586910 TI - Cosmology. MAP glimpses universe's rambunctious childhood. PMID- 12586909 TI - Gene therapy. RAC hears a plea for resuming trials, despite cancer risk. PMID- 12586911 TI - Astronomy. Institute faulted on attitudes toward women. PMID- 12586912 TI - Russian science. Academy plucks best biophysicists from a sea of mediocrity. PMID- 12586913 TI - European Union research. Ethics group gives qualified nod to placebos. PMID- 12586914 TI - RNA interference. Mini RNA molecules shield mouse liver from hepatitis. PMID- 12586916 TI - Space shuttle. After Columbia, a new NASA? PMID- 12586915 TI - Agriculture. Study shows richer harvests owe much to climate. PMID- 12586917 TI - Space shuttle. Disaster sets off science scramble. PMID- 12586918 TI - Space shuttle. Columbia disaster underscores the risky nature of risk analysis. PMID- 12586919 TI - Medicine. Tracing the steps of metastasis, cancer's menacing ballet. PMID- 12586920 TI - Medicine. A clash over genes that foretell metastasis. PMID- 12586921 TI - Genomics. Tinker, tailor: can Venter stitch together a genome from scratch? PMID- 12586922 TI - More on the animal rights debate. PMID- 12586923 TI - Caution in China over GM crops. PMID- 12586924 TI - Cerebral hemorrhage and amyloid-beta. PMID- 12586926 TI - Comment on "Determination of deforestation rates of the world's humid tropical forests". PMID- 12586927 TI - Science and the law. Patent swords and shields. PMID- 12586928 TI - Science and the law. Working through the patent problem. PMID- 12586929 TI - Neuroscience. A measured look at neuronal oxygen consumption. PMID- 12586930 TI - Biochemistry. How iron activates O2. PMID- 12586932 TI - Geoscience. Microbial geoengineers. PMID- 12586931 TI - Signal transduction. Imposing specificity on kinases. PMID- 12586933 TI - Archaeology. Invisible clues to New World plant domestication. PMID- 12586934 TI - Immunology. Regulating the regulators. PMID- 12586935 TI - Climate and management contributions to recent trends in U.S. agricultural yields. PMID- 12586936 TI - Crystallographic and spectroscopic characterization of a nonheme Fe(IV)-O complex. AB - Following the heme paradigm, it is often proposed that dioxygen activation by nonheme monoiron enzymes involves an iron(IV)=oxo intermediate that is responsible for the substrate oxidation step. Such a transient species has now been obtained from a synthetic complex with a nonheme macrocyclic ligand and characterized spectroscopically. Its high-resolution crystal structure reveals an iron-oxygen bond length of 1.646(3) angstroms, demonstrating that a terminal iron(IV)=oxo unit can exist in a nonporphyrin ligand environment and lending credence to proposed mechanisms of nonheme iron catalysis. PMID- 12586938 TI - Low-noise current amplifier based on mesoscopic Josephson junction. AB - We used the band structure of a mesoscopic Josephson junction to construct low noise amplifiers. By taking advantage of the quantum dynamics of a Josephson junction, i.e., the interplay of interlevel transitions and the Coulomb blockade of Cooper pairs, we created transistor-like devices, Bloch oscillating transistors, with considerable current gain and high-input impedance. In these transistors, the correlated supercurrent of Cooper pairs is controlled by a small base current made up of single electrons. Our devices reached current and power gains on the order of 30 and 5, respectively. The noise temperature was estimated to be around 1 kelvin, but noise temperatures of less than 0.1 kelvin can be realistically achieved. These devices provide quantum-electronic building blocks that will be useful at low temperatures in low-noise circuit applications with an intermediate impedance level. PMID- 12586937 TI - Crystal structure of naphthalene dioxygenase: side-on binding of dioxygen to iron. AB - Binding of oxygen to iron is exploited in several biological and chemical processes. Although computational and spectroscopic results have suggested side on binding, only end-on binding of oxygen to iron has been observed in crystal structures. We have determined structures of naphthalene dioxygenase that show a molecular oxygen species bound to the mononuclear iron in a side-on fashion. In a complex with substrate and dioxygen, the dioxygen molecule is lined up for an attack on the double bond of the aromatic substrate. The structures reported here provide the basis for a reaction mechanism and for the high stereospecificity of the reaction catalyzed by naphthalene dioxygenase. PMID- 12586939 TI - A sublimation model for martian south polar ice features. AB - In their pioneering work, Leighton and Murray argued that the Mars atmosphere, which at present is 95% carbon dioxide, is controlled by vapor equilibrium with a much larger polar reservoir of solid carbon dioxide. Here we argue that the polar reservoir is small and cannot function as a long-term buffer to the more massive atmosphere. Our work is based on modeling of the circular depressions commonly found on the south polar cap. We argue that a carbon dioxide ice layer about 8 meters thick is being etched away to reveal water ice underneath. This is consistent with thermal infrared data from the Mars Odyssey mission. PMID- 12586940 TI - Phytolith evidence for early Holocene Cucurbita domestication in southwest Ecuador. AB - Cucurbita (squash and gourd) phytoliths recovered from two early Holocene archaeological sites in southwestern Ecuador and directly dated to 10,130 to 9320 carbon-14 years before the present (about 12,000 to 10,000 calendar years ago) are identified as derived from domesticated plants because they are considerably larger than those from modern wild taxa. The beginnings of plant husbandry appear to have been preceded by the exploitation of a wild species of Cucurbita during the terminal Pleistocene. These data provide evidence for an independent emergence of plant food production in lowland South America that was contemporaneous with or slightly before that in highland Mesoamerica. PMID- 12586941 TI - Taming of a poison: biosynthesis of the NiFe-hydrogenase cyanide ligands. AB - NiFe-hydrogenases have an Ni-Fe site in which the iron has one CO and two CN groups as ligands. Synthesis of the CN ligands requires the activity of two hydrogenase maturation proteins: HypF and HypE. HypF is a carbamoyltransferase that transfers the carbamoyl moiety of carbamoyladenylate to the COOH-terminal cysteine of HypE and thus forms an enzyme-thiocarbamate. HypE dehydrates the S carbamoyl moiety in an adenosine triphosphate-dependent process to yield the enzyme thiocyanate. Chemical model reactions corroborate the feasibility of this unprecedented biosynthetic route and show that thiocyanates can donate CN to iron. This finding underscores a striking parallel between biochemistry and organometallic chemistry in the formation of an iron-cyano complex. PMID- 12586942 TI - Single-neuron activity and tissue oxygenation in the cerebral cortex. AB - Blood oxygen level-dependent functional magnetic resonance imaging uses alterations in brain hemodynamics to infer changes in neural activity. Are these hemodynamic changes regulated at a spatial scale capable of resolving functional columns within the cerebral cortex? To address this question, we made simultaneous measurements of tissue oxygenation and single-cell neural activity within the visual cortex. Results showed that increases in neuronal spike rate were accompanied by immediate decreases in tissue oxygenation. We used this decrease in tissue oxygenation to predict the orientation selectivity and ocular dominance of neighboring neurons. Our results establish a coupling between neural activity and oxidative metabolism and suggest that high-resolution functional magnetic resonance imaging may be used to localize neural activity at a columnar level. PMID- 12586943 TI - Contrast tuning in auditory cortex. AB - The acoustic features useful for converting auditory information into perceived objects are poorly understood. Although auditory cortex neurons have been described as being narrowly tuned and preferentially responsive to narrowband signals, naturally occurring sounds are generally wideband with unique spectral energy profiles. Through the use of parametric wideband acoustic stimuli, we found that such neurons in awake marmoset monkeys respond vigorously to wideband sounds having complex spectral shapes, preferring stimuli of either high or low spectral contrast. Low contrast-preferring neurons cannot be studied thoroughly with narrowband stimuli and have not been previously described. These findings indicate that spectral contrast reflects an important stimulus decomposition in auditory cortex and may contribute to the recognition of acoustic objects. PMID- 12586944 TI - Dolly, the so-called clone. AB - The word 'clone' is in the forefront of scientific terms that are familiar to the general public. But what exactly does it mean? Suprisingly, there is no simple answer, for different scientists are using the term for different entities. PMID- 12586945 TI - Toxic residues? PMID- 12586946 TI - Being definite. PMID- 12586948 TI - Harder than wiggling one's ears. PMID- 12586947 TI - Biologists on the loose. PMID- 12586949 TI - No comment - outdoor reared pork sausages. PMID- 12586951 TI - Obesity: part one - the greatest health threat facing mankind. AB - 'In the 15 minutes it will take you to read this article nine people in the USA, and one person in the UK, will have died as a direct consequence of obesity related illnesses.' PMID- 12586950 TI - The spark of life. AB - The sperm activates the egg to begin developing into an embryo by triggering a wave of intracellular calcium release, which sweeps across the egg like a forest fire. The identity of the signaling molecules that mediate this fundamental event has remained a mystery until recently. Now, new information emerging from the genome projects has made it possible to isolate and characterize the proteins responsible. PMID- 12586952 TI - Sinking methane. AB - Concentrations of the powerful greenhouse gas, methane, in our atmosphere have doubled since the beginning of the industrial age. Reducing these levels is a vital part of global efforts to combat global warming. Could we make use of the Earth's own methane sinks? PMID- 12586953 TI - Re-introduction of birds and mammals to the British Isles. AB - Large and exciting raptors like white-tailed eagle and red kite have been restored to the skies of Scotland and England, and now there are advanced plans to return the beaver. These successes encourage debate on other missing animals, with the elusive Eurasian lynx as the next best candidate. PMID- 12586954 TI - The Bloom's syndrome helicase: keeping cancer at bay. AB - Bloom's syndrome, a very rare inherited disorder, predisposes its sufferers to the full range of cancers that afflict humanity. This predisposition is rooted in just one defective gene on chromosome 15. It encodes the BLM helicase - an enzyme that ordinarily protects against DNA damage arising during replication. PMID- 12586955 TI - Ripe for a change. AB - As rich sources of vitamins and minerals, fruits are an important part of our diet. However, they are highly perishable. As the biochemical basis for fruit quality is being unraveled, so genetic approaches are being developed to modify the way in which fruit ripen, for the benefit of the consumer. PMID- 12586956 TI - Insect repellants in urban settings. AB - Why kill bugs when you can just make them go away? Used primarily for biting insect control, repellants may control household pests as well, once their potential is fully realized. Repellents may provide low-dose, specific and low toxicity augmentations to conventional pesticides applied around the home and workplace. PMID- 12586957 TI - Biogeography, a dirty word? AB - The newly developing fields of phylogeography and macroecology are ignoring historical patterns at their peril. PMID- 12586958 TI - Local anesthetics induce human renal cell apoptosis. AB - Renal cell apoptosis contributes significantly to the pathogenesis of acute renal failure. Local anesthetics induce apoptosis in neuronal and lymphocytic cell lines. We examined the effects of chronic (48 h) local anesthetic treatment (lidocaine, bupivacaine and tetracaine) on human proximal tubular (HK-2) cells. Apoptosis induction was assessed by detecting poly(ADP)-ribose polymerase fragmentation, caspase activation, terminal deoxynucleotidyl transferase biotin dUTP nick end labeling (TUNEL) staining, DNA laddering and by cellular morphology. Cell death was quantified by measuring neutral red dye uptake and lactate dehydrogenase released into the cell culture medium. All 3 local anesthetics caused concentration-dependent cell death, induced HK-2 cell apoptosis and potentiated TNF-alpha induced apoptosis. Local anesthetics induced HK-2 cell apoptosis by activation of caspases 3, 6, 7, 8 and 9. ZVAD-fmk, a pan caspase inhibitor, blocked the local anesthetic induced HK-2 cell apoptosis. Local anesthetics also inhibited the activities of anti-apoptotic kinases protein kinase B (Akt) and extracellular signal regulated mitrogen-activated protein kinase. Local anesthetic's pro-apoptotic effects are independent of sodium channel inhibition as tetrodotoxin, a selective voltage-gated sodium channel blocker, failed to mimic local anesthetic-mediated induction or potentiation of HK-2 cell apoptosis. We conclude that local anesthetics induce human renal cell apoptotic signaling by caspase activation and via inhibition of pro-survival signaling pathways. PMID- 12586959 TI - Depressive personality and its relationship to depressed mood, interpersonal loss, negative parental perceptions, and perfectionism. AB - Eighty veteran psychiatric outpatients were evaluated for depressive personality disorder on the Depressive Personality Disorder Inventory (DPDI). It was predicted that those classified with depressive personality would report higher levels of interpersonal loss, negative perceptions of their parents, and higher levels of perfectionism than psychiatric control subjects. Nine of the 12 measures of these variables were significantly greater in those with depressive personality compared with psychiatric control subjects. When statelike depression was controlled for, seven of the nine variables still significantly differed between the two groups. Hierarchical regression analysis and discriminant function analysis found that these variables predicted 9% of the variance in the DPDI above and beyond statelike depression, and that a combination of these variables correctly classified 91% of the depressive personalities and 88% of the psychiatric control subjects. It is concluded that, as hypothesized, depressive personality disorder is associated with loss, negative parental perceptions, and perfectionism, and that these relationships are not accounted for exclusively by a depressed mood. PMID- 12586960 TI - Personality dimensions in schizophrenia: associations with symptoms and coping. AB - While individual differences in personality exist among persons with schizophrenia and predate the onset of illness, less is known about their relationship to outcome. This study examined whether levels of three personality dimensions-neuroticism, extraversion, and agreeableness-are associated with symptomatology and coping in persons with schizophrenia. Symptom, personality, and coping measures were obtained for 59 participants with schizophrenia. Personality and coping measures were obtained for 17 persons in a community comparison group. Relative to the community comparison groups, participants with schizophrenia had higher levels of neuroticism, lower levels of extraversion and agreeableness, and tended to employ more avoidant styles of coping. Participants with schizophrenia who had higher levels of neuroticism had greater positive and emotional discomfort symptoms and greater preferences for avoidant coping strategies. Participants with schizophrenia who had higher levels of agreeableness had lower levels of positive and excitement symptoms. No links were found between extraversion and outcome measures. Implications for understanding how personality may affect outcome are discussed. PMID- 12586961 TI - Initial and final work performance in schizophrenia: cognitive and symptom predictors. AB - Studies have shown that cognitive functioning may limit the rate, tenure, and type of work performed by people with schizophrenia. The present study tested the hypothesis that cognitive abilities needed for initial improvement in work performance would differ from those needed in later vocational development. Ninety-six outpatients with schizophrenia or schizoaffective disorder who participated in a work rehabilitation program were administered neuropsychological testing at intake. Their work performance was evaluated biweekly for 26 weeks. Cognitive test variables were entered into regressions predicting the slope of individual performance curves from weeks 1 to 13 and 13 to 26. Neuropsychological variables accounted for 28% of the variance in slope during the initial period. The strongest predictor was a Continuous Performance Task variable that measures inattentiveness. Other important variables were measures of idiosyncratic thinking, cognitive flexibility, and verbal memory. Neuropsychological variables accounted for 19% of the variance in the slope during the final period. The strongest predictor was verbal learning, while measures of cognitive impulsivity and psychomotor functioning were also significant contributors. There was no relationship between work performance and symptoms for weeks 1 to 13 or 13 to 26. These findings suggest that while attention is more important for initial success, verbal memory becomes more important for sustained improvement. Remediating or accommodating such deficits in a time-sensitive fashion may be a necessary feature for successful rehabilitation. PMID- 12586962 TI - Comorbidity as a predictor of symptom change after treatment in combat-related posttraumatic stress disorder. AB - Posttraumatic stress disorder (PTSD) is a difficult condition to treat, and existing studies show considerable variability in outcome. Investigations of factors that influence outcome have the potential to inform alternate treatment approaches to maximize benefits gained from interventions for the disorder. Because PTSD is commonly associated with comorbidity, it is important to investigate the influence of comorbidity on symptom change after treatment. This article examines pretreatment and 9-month follow-up data for 134 Australian Vietnam veterans who attended a treatment program for combat-related PTSD. A series of analyses were conducted to investigate the influence of the comorbid factors of anxiety, depression, anger, and alcohol use on PTSD symptom change after treatment. Analyses identified anger, alcohol, and depression as significant predictors of symptom change over time, independent of the effect of initial PTSD severity. Further analyses indicated that anger at intake was the most potent predictor of symptom change. Further investigations of anger as an influence on symptom change after treatment of combat-related PTSD is recommended. PMID- 12586963 TI - The TAFES multi-family group intervention for Kosovar refugees: a feasibility study. AB - The object of this study was to describe a feasibility study of the Tea and Families Education and Support (TAFES) intervention used in a group of newly resettled adult refugees from Kosova. The subjects were 86 newly resettled Kosovar refugees in Chicago who gave informed consent to participate in an investigation of the TAFES intervention. All subjects received family home visits, and most participated in the TAFES multi-family groups. The instruments were administered to adult participants before and 3 months after the intervention. The TAFES program had contact with 61 Kosovar refugee families, of which 42 families (69%) engaged in TAFES groups, including families with educated and working members. Several characteristics were associated with engaging in TAFES groups and included lower monthly family income and higher age of the first child. The uncontrolled postintervention assessments demonstrated increases in social support and psychiatric service use associated with engagement in the TAFES group. Participants also showed time changes in scale scores assessing trauma mental health knowledge, trauma mental health attitudes, and family hardiness. This study provides preliminary evidence that multi-family support and education groups are a feasible and possibly beneficial intervention for newly resettled refugees and indicates the need for further studies. PMID- 12586964 TI - Stress symptoms among African-American college students after the September 11, 2001 terrorist attacks. AB - The current study examined stress reactions to the events of September 11, 2001 among African-American college students not directly exposed to the attacks. Within 3 days of September 11, 219 undergraduates (78.3% women) completed self report measures assessing stress symptoms and other reactions to the attacks. The results indicated that many students experienced a variety of stress symptoms and distressing thoughts and feelings in response to the events of September 11, including academic problems, concerns about family and friends in the military, and fear about war. Most students were highly distressed by specific attack related news reports and images. Anger toward persons of Middle Eastern descent was not frequently reported. Later college year and having parents not currently together were predictors of overall stress symptom severity as assessed by the Posttraumatic Stress Disorder Checklist. Later college year also predicted academic problems after September 11. The findings are discussed in terms of intervention implications and suggested directions for future research. PMID- 12586966 TI - Mass hysteria and perceptions of the supernatural among adolescent girl students in Taiwan. PMID- 12586965 TI - Prevalence of dissociative experiences in a community sample: relationship to gender, ethnicity, and substance use. AB - This study examined the relationship between gender, ethnicity, substance use, and dissociation in a community sample of adults. Telephone interviews were conducted using, among others, measures of dissociation (Dissociative Experiences Scale-Taxon [DES-T]) and alcohol use (Alcohol Use Disorders Identification Test [AUDIT]) to screen a representative sample of 1007 adults in Memphis, Tennessee. Six percent of respondents endorsed four or more lifetime dissociative symptoms, whereas approximately one third of respondents endorsed at least one symptom. Only 2% were identified as having pathological dissociation on the DES-T. DES-T scores were significantly associated with gender (male), ethnicity (African American), and harmful alcohol use. Dissociative symptoms appear to be relatively common in the population, but only a small proportion of individuals suffer from significant levels of dissociation. Further assessment of the link between demographic factors and the severity and expression of dissociative phenomena in nonclinical samples is warranted. PMID- 12586967 TI - Quality of life and family history in pathological gambling. PMID- 12586968 TI - Reactions of psychiatric and medical inpatients to terror and security instability in Israel. PMID- 12586969 TI - Perceived patient threat and neuroleptic dosage. PMID- 12586970 TI - Acute otitis media: more trouble with the evidence. PMID- 12586971 TI - Community outbreak of perianal group A streptococcal infection in Denmark. AB - BACKGROUND: Perianal group A streptococcal infection (PASI) occurs primarily in children. There is limited information on the incidence, transmission and treatment of PASI. We report a cluster of cases connected to a Danish kindergarten and observations of the incidence of PASI in the local population. SETTING: A Danish rural community with 1765 children 15 years and younger registered with two general practice clinics. METHODS: After being alerted of a possible cluster of PASI cases, all isolates of group A beta-hemolytic streptococci were collected and subjected to T typing and pulsed field gel electrophoresis (PFGE) if grown from either a rectal swab or an accompanying throat swab obtained in the offices of local general practitioners during the ensuing 4-month period. Clinical data were obtained from the files of the local general practitioners. RESULTS: Twelve cases of PASI were caused by group A beta hemolytic streptococci T type 28 with an identical PFGE profile: 6 of the cases were in children attending the same kindergarten, 4 were connected otherwise to the cluster and 2 cases seemed to be unrelated. Five cases of PASI with different T types and PFGE profiles were diagnosed during the same period giving an estimated annual incidence of 2 to 7 per 1000 children. Penicillin V was ineffective in 3 cases, and no recurrence was seen after change of the treatment to oral clarithromycin. CONCLUSIONS: A clone of T type 28 seemed to be the cause of the largest cluster of PASI cases described thus far. Clarithromycin was effective as second line treatment. An estimated annual baseline incidence of 2 to 7 per 1000 in the local population indicates that PASI may not be as rare as previously estimated. PMID- 12586972 TI - Cefepime use in a pediatric intensive care unit reduces colonization with resistant bacilli. AB - BACKGROUND: Cefepime has activity against many hospital-acquired Gram-negative pathogens resistant to earlier beta-lactam antibiotics. This study was designed to test whether preferential use of cefepime in a pediatric intensive care unit could reduce enteric colonization with antibiotic-resistant Gram-negative rods. METHODS: After a 6-month period of uncontrolled antibiotic use, cefepime was preferentially used during 2 years as treatment for nosocomial or serious community-acquired infection. Rectal swab specimens were obtained daily on every patient regardless of antibiotic exposure during the 6 months of uncontrolled antibiotic use and during the first and last 6 months of the 2 years of cefepime preference. The study outcome was rectal colonization with a facultative Gram negative rod resistant to at least one of four antibiotics: cefepime; ceftazidime; gentamicin; or piperacillin-tazobactam. RESULTS: The incidence of colonization by a resistant organism decreased only slightly during the first 6 months of cefepime use. By contrast, the number of antibiotic-resistant bacilli isolated from rectal swab specimens diminished from 27.6/100 patients during the baseline period to 12.9/100 patients by the last 6 months of the 2 years of cefepime preference (P < 0.01). The proportion of patients harboring at least one resistant organism decreased from 11.6% to 7.4% during the same time period (P < 0.01). A decrease in colonization with resistant organisms occurred for all the tested resistance phenotypes, including cefepime. CONCLUSION: Cefepime may possess a low potential for promoting bacillary resistance in critically ill patients, suggesting that its preferential use might be a key element in limiting the presence of antibiotic resistance in the intensive care unit. PMID- 12586973 TI - Absence of associations between influenza-associated encephalopathy and human herpesvirus 6 or human herpesvirus 7. AB - BACKGROUND: Influenza-associated encephalopathy is a severe complication of influenza virus infection, but its pathogenesis is unknown. It was recently suggested that the neurologic complications of influenza, including encephalopathy, are associated with human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7). AIM To confirm or refute the association between influenza associated encephalopathy and HHV-6 or HHV-7. METHODS: Cerebrospinal fluid and serum from 25 patients with central nervous system complications of influenza (18 patients with encephalopathy and 7 patients with febrile convulsions) were investigated. The specimens were examined by real time polymerase chain reaction (PCR) and nested PCR for HHV-6 and HHV-7 DNA. RESULTS: In the cerebrospinal fluid samples neither HHV-6 DNA nor HHV-7 DNA was detected by real time PCR or nested PCR. HHV-6 DNA was detected in a single serum sample from a patient with febrile convulsions. CONCLUSION: In our study there was no association with HHV-6 or HHV 7 in most patients with central nervous system complications of influenza. PMID- 12586974 TI - Thrombocytopenia after immunization of Canadian children, 1992 to 2001. AB - BACKGROUND: Thrombocytopenia occasionally follows immunization of children, especially after administration of measles-containing vaccines. The purpose of this study was to describe the clinical features of postimmunization thrombocytopenia, with emphasis on the rate of complications and outcome. METHODS: A prospective survey was conducted by 12 pediatric centers in Canada during 1992 to 2001. At each center a nurse monitor searched for inpatient cases. Cases were defined as having onset of clinical signs or laboratory measures of thrombocytopenia (platelet count, <50 x 10(9)/l) within 30 days after immunization. Cases were described in a standardized manner, including follow-up data as available. RESULTS: Sixty-one cases were detected, an average of 6 per year or approximately 1 case per 15,000 general hospital admissions. Median age of cases was 13 months. The mean platelet count at diagnosis was 8.6 x 10(9)/l. Most cases (79%) followed measles-containing vaccines. Only 1 child had a serious (fatal) complication. Platelet counts returned to normal within 30 days of onset in 46 of 57 children (80.7%) with information available. Five children (8.2%) had persistent or intermittent thrombocytopenia for 3 months or more. CONCLUSION: Thrombocytopenia associated with routine immunization of children is rare and usually benign, resolving within 1 month in most children. PMID- 12586975 TI - Antibody response to postexposure prophylaxis in infants born to hepatitis B surface antigen-positive women. AB - BACKGROUND: Annually 20,000 infants are born to hepatitis B surface antigen (HBsAg)-positive US women. Without prophylaxis 30% risk chronic hepatitis B virus infection, and 25% of those risk dying from resulting liver cirrhosis or liver cancer as adults. METHODS: We attempted to interview each HBsAg-positive pregnant woman reported to the health department between 1992 and 1997, to provide their infants with immunoprophylaxis at birth and in the clinic or home and to serotest at 9 to 15 months of age. RESULTS: Of 879 women reported, 92% enrolled; 787 delivered 796 live infants; 91% of infants received hepatitis B immunoglobulin; 98, 95 and 89% received hepatitis B vaccine (HepB) Doses 1, 2 and 3, respectively; and 80% were serotested. Of these 2.2% were HBsAg-positive and 97% had antibody to HBsAg (anti-HBs) of > or =10 mIU/ml. Anti-HBs concentrations measured in 504 infants were 10 to 99 mIU/ml (25%), 100 to 999 mIU/ml (43%) and > or =1000 mIU/ml (29%). Serotesting was less likely among infants of mothers <20 years of age [odds ratio (OR) 2.5]; white, non-Hispanic (OR 2.8); or with a household income of <$15,000/year (OR 2.0). Lower antibody titers were found when serotesting at 4 to 12 months than at <4 months after HepB-3 (OR 1.8 to 4.4), with HepB-3 receipt <6 months after HepB-2 (OR 2.5) and when household income was <$15,000/year (OR 2.1). CONCLUSIONS: Centralized case management with home visits resulted in high rates of complete immunoprophylaxis and postvaccination testing among infants born to HBsAg-positive women. Perinatal immunoprophylaxis was immunogenic under routine public health use, with higher anti-HBs titers occurring in infants tested <4 months postvaccination. Because infants in households with low income had higher rates of nonprotective antibody responses, they may benefit from extra efforts to ensure that serotesting is conducted postvaccination. PMID- 12586976 TI - Roxithromycin treatment of scrub typhus (tsutsugamushi disease) in children. AB - BACKGROUND: Although chloramphenicol and doxycycline have been used for the treatment of scrub typhus, a difficulty exists in determining which drug to use in treating children because of such potential complications as aplastic anemia or tooth discoloration. We evaluated the effect of roxithromycin, a macrolide antibiotic, on scrub typhus in children. METHODS: A retrospective analysis was conducted on 39 children with scrub typhus who were treated with doxycycline (DC), chloramphenicol (CM) or roxithromycin (RM) between 1991 and 2000. We divided the patients into the DC-treated group (DC group; 16 children), CM treated group (CM group; 14 children) and RM- treated group (RM group; 9 children) and compared these groups. RESULTS: Most cases (97%) occurred in October and November. Fever and rash were observed in all 39 cases, and an eschar was noted in 36 cases (92%). No statistical differences could be found between the 3 groups in mean age, duration of fever before admission, white blood cell (WBC) count and complications including abnormal liver enzymes. In most cases defervescence after treatment was within 24 h (34 cases, 87%) and during 24 to 48 h in 2 cases in the DC group, 1 case in the CM group and 2 cases in the RM group (no statistical difference). CONCLUSION: Roxithromycin was as effective as conventional doxycycline or chloramphenicol in children with scrub typhus and may be safer to use. PMID- 12586977 TI - Report from a WHO working group: standard method for detecting upper respiratory carriage of Streptococcus pneumoniae. AB - BACKGROUND: Numerous studies evaluating the efficacy of conjugate pneumococcal vaccines are being conducted or planned throughout the world. Some of these studies are evaluating the effect of vaccine on nasopharyngeal (NP) carriage. METHODS: The World Health Organization established a Working Group composed of representatives from these trials and other NP colonization experts to establish core, standardized methods for the study of pneumococcal NP colonization that could be used in these trials. The intent was to reduce or eliminate variability in key methods which themselves could contribute to variability of observed pneumococcal NP colonization. In this way variability of vaccine effects between trials on NP colonization could more easily be analyzed for population or vaccine differences without the confounding effect caused by differences in study methodology. RESULTS: This paper presents the evidence base supporting the need for standardized NP colonization study methods, the methods themselves (Core Consensus Methods found in the electronic version of this article at www.pidj.com and on the WHO website at http://www.who.int/vaccines-research/rd/docsrd.shtml), including collection techniques, culture media, equipment, serotyping, storage of specimens and transport of isolates agreed on by the Working Group as well as a discussion of research priorities. CONCLUSIONS: The Core Consensus Methods provide a common methodology to conduct pneumococcal NP colonization studies with minimum interstudy method variability. The intention is to allow more meaningful comparisons of study results from conjugate pneumococcal vaccine trials. PMID- 12586978 TI - Greater antibody responses to an eleven valent mixed carrier diphtheria- or tetanus-conjugated pneumococcal vaccine in Filipino than in Finnish or Israeli infants. AB - BACKGROUND: Antibody responses to pneumococcal conjugate vaccines may vary when administered in different populations or epidemiologic settings. Understanding the causes and significance of this variation could help to determine the number of doses and timing required for protection against pneumococcal diseases in each country. METHODS: This report compares antibody responses to aluminum-adjuvanted and nonadjuvanted mixed carrier 11-valent diphtheria- or tetanus-conjugated pneumococcal vaccine (11-PncTD) formulations when given at 6, 10 and 14 weeks and 9 months of age to Filipino infants (n = 51) and at 2, 4, 6 and 12 months of age to Finnish (n = 127) and Israeli (n = 124) infants. The study populations differ in their natural exposure to pneumococcus and risk factors for pneumococcal carriage and disease. RESULTS: Filipino and Israeli infants had slightly but significantly higher prevaccination geometric mean concentration (GMC) of antibodies than did the Finnish infants. After three doses of aluminum-adjuvanted 11-PncTD vaccine, the Filipino infants had 1.8 to 6.7 and 1.5 to 5.0 times higher GMC than the Finnish and Israeli infants, respectively, against pneumococcal serotypes conjugated to tetanus protein. The GMC of serotypes conjugated to diphtheria toxoid was 1.3 to 3.0 and 0.7 to 2.0 times the GMC in Finnish and Israeli infants, respectively. The nonadjuvanted vaccine formulation induced generally lower GMCs. CONCLUSIONS: The antibody responses to the tetanus conjugated polysaccharides were considerably higher in the Filipino than in the Finnish or Israeli infants. This may be a result of several factors including the priming effect of tetanus toxoid given to pregnant women, early pneumococcal nasopharyngeal acquisition and genetic differences among populations. PMID- 12586979 TI - Incidence and clinical features of hospitalization because of respiratory syncytial virus lower respiratory illness among children less than two years of age in a rural Asian setting. AB - BACKGROUND: Lower respiratory illness is the leading cause of child death in the developing world. Despite this few reports on respiratory syncytial virus (RSV) lower respiratory illness disease burden exist from rural areas of the developing world, and none exist for Indonesia. METHODS: We evaluated children living in any of 83 villages on Lombok Island, Indonesia who were <2 years of age when hospitalized for severe lower respiratory illness during 2000 and 2001. All hospitals on Lombok were included in the evaluation. We obtained the number of births and deaths that occurred within the study villages and time frame, allowing for incidence determination. RESULTS: Of 2677 children hospitalized for severe lower respiratory illness whose RSV status was determined, 23% had a positive test; this percentage varied from 50% at the end of the rainy season to 0% shortly before the start of the rainy season. Among children <2 years of age, the confirmed and estimated incidences of severe RSV lower respiratory illness hospitalization were 10 and 14 per 1000 child-years, respectively; values for children <1 year of age were 17 and 25 per 1000 child-years. The confirmed case fatality percentage among RSV-positive cases was 1.9%; however, 84% of children who died were dead before RSV status could be determined, suggesting the actual case fatality percentage was higher. CONCLUSIONS: Lombok has a large burden of severe childhood RSV lower respiratory illness, and death occurs frequently. Novel RSV vaccines thus could have a substantial positive impact on lower respiratory illness morbidity and mortality. PMID- 12586980 TI - Long term antibody response to hepatitis B vaccination beginning at birth and to subsequent booster vaccination. AB - BACKGROUND: Few studies have examined the long term persistence of antibody after hepatitis B immunization beginning at birth and the response to a subsequent challenge with a booster dose of vaccine. METHODS: Two groups of children received hepatitis B vaccine on a schedule of birth and 1 and 6 months of age. Group 1 received recombinant vaccine and a booster dose at 5 years of age. Group 2 received plasma-derived vaccine and a booster dose at 9 years of age. Group 1 children were tested for antibody after the primary vaccine series. All children were tested for antibody before administration of the booster dose and at 2 and 4 weeks and 1 year after the booster. In addition all children were tested for markers of hepatitis B virus infection. RESULTS: Antibody testing conducted after the primary series for children in Group 1 (n = 70) showed that 90% had protective antibody concentrations at 13 months of age, and testing before the booster dose showed that 41% had protective antibody concentrations. All children with protective antibody concentrations after the primary series had an anamnestic antibody response to the booster dose. In Group 2 (n = 41) 39% of children had protective antibody concentrations before the booster dose, and 93% had an anamnestic antibody response to the booster dose. One year after the booster dose there were 26-fold and 11-fold declines in antibody concentration in Groups 1 and 2, respectively. CONCLUSIONS: A primary vaccination series with either plasma-derived or recombinant hepatitis B vaccine affords long term protection for children when vaccinated beginning soon after birth. PMID- 12586981 TI - Influenza diagnosis and treatment in children: a review of studies on clinically useful tests and antiviral treatment for influenza. AB - BACKGROUND: Prompt testing for influenza can help guide clinical management of patients with suspected influenza. Three antiviral medications, amantadine, oseltamivir and zanamivir, are approved for treatment of influenza in children. Rimantadine and ribavirin have also been used. OBJECTIVES: To review the published evidence on clinically useful diagnostic tests and antiviral treatment for influenza virus infections in children. METHODS: Studies published from 1966 through September 2002 were reviewed on clinical diagnosis, immunofluorescence and rapid influenza tests and on antiviral treatment of influenza virus infections among pediatric populations. RESULTS: No studies assessed the accuracy of clinical diagnosis of influenza in children compared with viral culture. Compared with viral culture, direct immunofluorescence antibody and indirect immunofluorescence antibody tests for influenza had fair to moderate median sensitivities and high median specificities, whereas rapid influenza diagnostic tests had moderate median sensitivities and moderately high median specificities. No randomized, placebo-controlled studies were found of amantadine or rimantadine for treatment of influenza A. In a few separate controlled studies, oseltamivir, zanamivir and ribavirin each reduced symptom duration of influenza compared with placebo. CONCLUSIONS: Additional data are needed about the accuracy of clinical diagnosis of influenza in children. Although direct immunofluorescence antibody staining, indirect immunofluorescence antibody staining and rapid tests are moderately to reasonably accurate in detecting influenza virus infections in children, physicians should use clinical judgment and local surveillance data about circulating influenza viruses when interpreting test results. Further controlled studies of the efficacy, adverse effects and emergence of antiviral resistance during treatment of influenza are needed for all of the antiviral drugs. PMID- 12586982 TI - Dengue virus infection. PMID- 12586983 TI - Neurocysticercosis. PMID- 12586984 TI - Agrobacterium radiobacter bacteremia in pediatric patients: case report and review. AB - Agrobacterium radiobacter is an opportunistic pathogen often associated with indwelling catheters. We report five children with central venous catheter associated A. radiobacter bacteremia and review the characteristics of pediatric Agrobacterium infections. Cure was achieved with appropriate antibiotics, often ticarcillin-clavulanate and gentamicin, and removal of the catheter. PMID- 12586985 TI - Intramedullary abscess of the spinal cord in children: a case report and review of the literature. AB - A case report of an intramedullary spinal cord abscess in a 13-month-old boy and a review of relevant existing pediatric literature is presented. Thirty-eight cases of pediatric intramedullary spinal cord abscess are analyzed for presenting signs and symptoms, microbiology of isolated organisms, surgical intervention, antibiotic administration and outcome. The most significant variable on outcome is timely surgical intervention, followed by appropriate antibiotic administration. PMID- 12586986 TI - Need for equivalence testing of efficacy of alternative antibiotics for treatment of pertussis. PMID- 12586987 TI - Report from a WHO Working Group: standard method for detecting upper respiratory carriage of Streptococcus pneumoniae. AB - BACKGROUND: Numerous studies evaluating the efficacy of conjugate pneumococcal vaccines are being conducted or planned throughout the world. Some of these studies are evaluating the effect of vaccine on nasopharyngeal (NP) carriage. METHODS: The World Health Organization established a Working Group comprised of representatives from these trials and other NP colonization experts to establish core, standardized methods for the study of pneumococcal NP colonization that could be used in these trials. The intent was to reduce or eliminate variability in key methods which themselves could contribute to variability of observed pneumococcal NP colonization. In this way variability of vaccine effects between trials on NP colonization could more easily be analyzed for population or vaccine differences without the confounding effect caused by differences in study methodology. RESULTS: This paper presents the evidence base supporting the need for standardized NP colonization study methods, the methods themselves (Core Consensus Methods), including collection techniques, culture media, equipment, serotyping, storage of specimens and transport of isolates agreed on by the Working Group as well as a discussion of research priorities. CONCLUSIONS: The Core Consensus Methods provide a common methodology to conduct pneumococcal NP colonization studies with minimum interstudy method variability. The intention is to allow more meaningful comparisons of study results from conjugate pneumococcal vaccine trials. PMID- 12586988 TI - Anesthesia technologies meet European standards. PMID- 12586989 TI - Guidelines for completing the Perioperative Anesthesia Record. PMID- 12586990 TI - Should disclosure of the danger of awareness during general anesthesia be a part of preanesthesia consent? AB - Awareness during anesthesia (AGA) is known as an intraoperative incident which could lead to a series of untoward effects, among them symptoms compatible with the posttraumatic stress syndrome (PTSS). Incidence of AGA ranges between 0.1% and 0.7%, most of the reports indicating a 0.2% rate of all general anesthesias. Nevertheless, some patients are considered to be in a higher than usual risk for developing this incident. The list of AGA high-risk situations include caesarian section, open heart surgical procedures, marked obesity, major trauma with hemodynamic instability and chronic use of drugs, alcohol or tobacco smoking. The usual preanesthetic informed consent does not mention AGA among the possible undesired effects of general anesthesia, neither in Israel nor in other parts of the world. This paper rises the question of the indication to discuss the AGA matter, as part of the informed consent, with any patient who is prone develop it in a significant higher percentage than the general population. The topic can be discussed by the primary care physician or by the surgeon, but this represents the obvious task of the anesthesiologist during his/her first contact with the patient before anesthesia and surgery. It is the authors belief that a preoperative discussion on AGA might substantially reduce the magnitude of repercussions of AGA among high-risk patients to develop this anesthetic complication. PMID- 12586991 TI - Quality control of sedation for diagnostic radiological procedures in paediatric patients (waiting for guidelines). AB - BACKGROUND: The number of children requiring sedation for radiological procedures is increasing. Anaesthesiologists are increasingly involved in giving sedation or general anaesthesia in the rooms of the Radiology Department. This activity is not easy, and can be dangerous. The procedure is often performed on an ambulatory basis, so the child must be alert and discharged rapidly after the procedure. METHODS: We reviewed the medical charts of 488 patients in order to evaluate the incidence of complications during deep sedation for diagnostic radiological procedures. The patients were sedated with intravenous thiopental or propofol, or with oral chloral hydrate. All the patients were breathing spontaneously and received only supplemental O(2). RESULTS: We found only a few cases of complications, immediately treated without any recourse to tracheal intubation: respiratory failure with arterial desaturation to 94%, regurgitation, vomiting and persistent cough. CONCLUSIONS: On the basis of our experience, we believe that deep sedation with endovenous drugs guarantees safety and rapid discharge after the procedure. PMID- 12586992 TI - [Percutaneous cervical cordotomy in the therapy of incidence pain]. AB - BACKGROUND: Incident pain does not respond to opioid treatment and it is not easily relieved with other therapeutic strategies (local intrapleural or spinal analgesia, phenol blocks etc.). For this reason cervical percutaneous cordotomy at C(1)-C(2) interspace is the only effective antalgic therapy in patients whose life expectancy is more than three to six months. METHODS: This study is a rectrospective review of 22 patients with cancer and incident pain from brachial, lumbar-sacral plexus injury and gluteal ulcer. RESULTS: Cordotomy provided excellent contralateral side pain relief in 21 patients; pain relief was maintained up to death and to the moment of last observation in living patients. In one deaf patient it was impossible to carry out the procedure due to incomplete co-operation and pain returned after 48 hours. Ventilatory depression caused death in one patient. Other complications recorded included ataxia, headache, motor deficit, dysesthesia and orthostatic hypotension. CONCLUSIONS: The conclusion is drawn that percutaneous cordotomy should, in carefully selected cases, be considered the only technique to relieve incident pain. PMID- 12586993 TI - Systemic capillary leak syndrome. Case report. AB - We here report the case of a patient with systemic capillary leak syndrome (SCLS). This syndrome is a rare condition characterized by recurrent episodes of hypotension with hemoconcentration and hypoproteinemia. It is due to unexplained episodic capillary hyperpermeabilty that results in fluid and protein shift from the intravascular to the interstitial space: generalized edema, shock and renal failure follow. A 59 yo man was admitted to our intensive care unit because of unexplained shock with hemoconcentration, renal failure, and metabolic acidosis. Previous attemps to reverse shock in a medical ward with crystalloids and dopamine failed. An abdominal CT scan, a TEE, and chest X ray study were inconclusive. No sign or history of major infections or anaphylaxis were present. The patient was resuscitated with massive fluid infusions and norepinephrine on the guide of a Swan Ganz catheter. The diagnosis was made on the basis of a previous episode of severe shock complicated with renal failure and a compartment syndrome, the hemoconcentration, and the negative cardiopulmonary findings. A small amount of monoclonal immunoglobulin G, kappa chain, found in the serum confirmed the diagnosis. The SCLS should be considered in the differential diagnosis of idiopathic and anaphylactic shock. Patients may benefit from a prophylactic treatment with theophilline and terbutaline. PMID- 12586994 TI - PPARgamma agonists in the treatment of type II diabetes: is increased fatness commensurate with long-term efficacy? AB - The nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the PPAR family. The endogenous activators of all members of the PPAR family are a variety of fatty acids, which suggests that the PPARs are highly involved in lipid metabolism. In the present paper, the current understanding of the involvement of PPARgamma in adipocyte proliferation and adipose tissue formation is extensively reviewed, and it is stressed that PPARgamma seems to be a major regulator in the differentiation of adipocytes. Thiazoledinediones (TZDs) are a group of PPARgamma-agonists used in the treatment of type 2 diabetes (T2D) since 1997. They are characterized by their ability to decrease insulin resistance, and have been suggested to slow down the progression of insulin resistance. Treatment with TZD requires several weeks of treatment to decrease plasma glucose levels, but in addition they markedly decrease plasma triglycerides and free fatty acids. A major drawback of treatment with TZD is body fat gain, but some evidence suggests that the fat is redistributed in a favourable direction, that is, from visceral to subcutaneous depots. However, the effect of long-term treatment on weight gain following TZD treatment is unknown, and it may be questioned whether the use of these 'adipogenic compounds' is appropriate, considering that excess body fat is almost a prerequisite for the development of type 2 diabetes. PMID- 12586995 TI - Early determinants of childhood overweight and adiposity in a birth cohort study: role of breast-feeding. AB - BACKGROUND: The prevalence of adiposity in childhood is increasing. Is breast feeding protective as suggested by cross-sectional studies? OBJECTIVE: In a longitudinal birth cohort study, we tested whether breast-feeding for more than 2 months has preventive effects against overweight and adiposity at 6 y. DESIGN: Of 1314 children representing the catchment areas of six delivery units, 918 could be followed up to the age of 6 y. Height, weight, and skin-fold thickness were measured at regular visits. As the criteria of overweight, obesity, and adiposity in the children, the 90th and the 97th percentiles of BMI and skin-fold values were used. Parents with a BMI at or above the 90th percentile, which was 27 kg/m(2) or more, were considered overweight. Infants bottle-fed from birth or breast-fed for less than 3 months were classified as 'bottle-fed' (BO), and those breast-fed for 3 months and more as 'breast-fed' (BR). Univariate comparisons and logistic regression analysis were performed applying SAS 6.12. The final logistic model consisted of the 480 cases for whom complete data for all variables were available. The potential effect of loss to follow-up was analysed by the Cochran Mantel-Haenzel test: the outcomes were not significantly influenced by loss to follow-up. RESULTS: At birth BMIs were nearly identical in both groups. By 3 months, BO had significantly higher BMIs and thicker skin folds than BR. From 6 months on, compared to BR, a consistently higher proportion of BO children exceeded the 90th and the 97th percentile of BMI and skin-fold thickness reference values. From the age of 4 y to 5 and 6 y, in BO the prevalence of obesity nearly doubled and tripled, respectively. With only minor changes of obesity prevalence in BR, the difference of BMI and skin-fold thickness between groups became statistically significant. Logistic regression analysis revealed that overweight of the mother, maternal smoking during pregnancy, bottle feeding, and low social status remained important risk factors for overweight and adiposity at 6 y of age. CONCLUSION: A maternal BMI of > or =27, bottle-feeding, maternal smoking during pregnancy, and low social status are risk factors for overweight and adiposity at 6 y of age. Early bottle-feeding brings forward the obesity rebound, predictive of obesity in later life. PMID- 12586996 TI - Neonatal anthropometry: the thin-fat Indian baby. The Pune Maternal Nutrition Study. AB - OBJECTIVE: To examine body size and fat measurements of babies born in rural India and compare them with white Caucasian babies born in an industrialised country. DESIGN: Community-based observational study in rural India, and comparison with data from an earlier study in the UK, measured using similar methods. SUBJECTS: A total of 631 term babies born in six rural villages, near the city of Pune, Maharashtra, India, and 338 term babies born in the Princess Anne Hospital, Southampton, UK. MEASUREMENTS: Maternal weight and height, and neonatal weight, length, head, mid-upper-arm and abdominal circumferences, subscapular and triceps skinfold thicknesses, and placental weight. RESULTS: The Indian mothers were younger, lighter, shorter and had a lower mean body mass index (BMI) (mean age, weight, height and BMI: 21.4 y, 44.6 kg, 1.52 m, and 18.2 kg/m(2)) than Southampton mothers (26.8 y, 63.6 kg, 1.63 m and 23.4 kg/m(2)). They gave birth to lighter babies (mean birthweight: 2.7 kg compared with 3.5 kg). Compared to Southampton babies, the Indian babies were small in all body measurements, the smallest being abdominal circumference (s.d. score: -2.38; 95% CI: -2.48 to -2.29) and mid-arm circumference (s.d. score: -1.82; 95% CI: -1.89 to -1.75), while the most preserved measurement was the subscapular skinfold thickness (s.d. score: -0.53; 95% CI: -0.61 to -0.46). Skinfolds were relatively preserved in the lightest babies (below the 10th percentile of birthweight) in both populations. CONCLUSIONS: Small Indian babies have small abdominal viscera and low muscle mass, but preserve body fat during their intrauterine development. This body composition may persist postnatally and predispose to an insulin resistant state. PMID- 12586997 TI - Physical activity in Filipino youth: the Cebu Longitudinal Health and Nutrition Survey. AB - OBJECTIVE: To describe physical activity (PA) and inactivity levels of Filipino youth aged 14-16 y. DESIGN: PA type and patterns were assessed in an adolescent cohort enrolled in the Cebu Longitudinal Health and Nutrition Survey. SUBJECTS: A total of 1053 males (aged 15.6+/-0.5 y; body mass index (BMI)=18.6+/-2.6 kg/m(2)) and 990 females (aged 14.6+/-0.5 y; BMI=18.8+/-2.4 kg/m(2)). MEASUREMENTS: PA was assessed by (1) self-reported usual PA (questionnaire); (2) 24-h objective monitoring (Caltrac accelerometer); and (3) 24 h recalls of PA during the objectively monitored time frame. RESULTS: As measured by Caltrac, males expended 378.0+/-164.5 kcal and females 271.0+/-105.4 kcal. The measurement approaches employed agreed in expected ways, given the limitations of each. Together the approaches used indicated that the primary sources of PA for this population are chores and active commuting. In all, 40% of the sample walked to school, less than 1% rode a bicycle, 22% rode in/on motorized transport, and 37% used some combination of walking and motorized transport. Further, gender differences exist for type, frequency, and duration of physical activities performed by Filipino youth. Specifically, females report engaging in few leisure-time physical activities and more household chores. In total, 48% of Filipino youth report watching > or =2 h/day of television; relatively few youth (7%) report watching >4 h/day at this time. An 18% of males report no vigorous activities compared to 87% of females. CONCLUSION: Clear gender differences in PA patterns and levels exist in Filipino adolescents. Nonsport/exercise type activities (eg chores and active commuting) currently provide a large proportion of daily activity for both males and females, however. It is anticipated that as modernization and technological advancement continue in this developing country, these primary sources of PA will decrease. PMID- 12586998 TI - Deficit in the discrimination of nonverbal emotions in children with obesity and their mothers. AB - OBJECTIVE: Referring to the alexithymia construct and Bruch's clinical observations, this study investigated the ability to decode nonverbal signs of emotion in obese boys and girls, and their mothers. METHOD: A group of 10 boys and 11 girls with obesity and their mothers, and a control group were tested. Both mothers and children were asked to recognize a set of 32 brief film sequences interpreted by four actors expressing four emotions (anger, sadness, fear, happiness) with two intensity levels. Each sequence was presented first without sound, second without video, and finally with video and sound. RESULTS: As expected, boys and girls suffering from obesity and their mothers showed a reduced ability to decode visual and verbal signs of emotion compared to the control group. DISCUSSION: This result may be interpreted in accordance with the alexithymia construct, and suggests the importance of developing therapeutic strategies to face alexithymic characteristics in obese children and their mothers. PMID- 12586999 TI - Relation between calcium intake and fat oxidation in adult humans. AB - OBJECTIVE: To determine if total calcium (Ca(2+)) intake and intake of Ca(2+) from dairy sources are related to whole-body fat oxidation. DESIGN: : Cross sectional study. SUBJECTS: A total of 35 (21 m, 14 f) non-obese, healthy adults (mean+/-s.d., age: 31+/-6 y; weight: 71.2+/-12.3 kg; BMI: 23.7+/-2.9 kg m(-2); body fat: 21.4+/-5.4%). MEASUREMENTS: Daily (24 h) energy expenditure (EE) and macronutrient oxidation using whole-room indirect calorimetry; habitual Ca(2+) intake estimated from analysis of 4-day food records; acute Ca(2+) intake estimated from measured food intake during a 24-h stay in a room calorimeter. RESULTS: Acute Ca(2+) intake (mg. kcal(-1)) was positively correlated with fat oxidation over 24 h (r=0.38, P=0.03), during sleep (r=0.36, P=0.04), and during light physical activity (r=0.32, P=0.07). Acute Ca(2+) intake was inversely correlated with 24-h respiratory quotient (RQ) (r=-0.36, P=0.04) and RQ during sleep (r=-0.31, P=0.07). After adjustment for fat mass, fat-free mass, energy balance, acute fat intake, and habitual fat intake, acute Ca(2+) intake explained approximately 10% of the variance in 24-h fat oxidation. Habitual Ca(2+) intake was not significantly correlated to fat oxidation or RQ. Total Ca(2+) intake and Ca(2+) intake from dairy sources were similarly correlated with fat oxidation. In backwards stepwise models, total Ca(2+) intake was a stronger predictor of 24 h fat oxidation than dairy Ca(2+) intake. CONCLUSION: Higher acute Ca(2+) intake is associated with higher rates of whole-body fat oxidation. These effects were apparent over 24 h, during sleep and, to a lesser extent, during light physical activity. Calcium intake from dairy sources was not a more important predictor of fat oxidation than total Ca(2+) intake. PMID- 12587000 TI - Cardiorespiratory fitness is associated with diminished total and abdominal obesity independent of body mass index. AB - OBJECTIVE: To determine whether, for a given body mass index (BMI), men and women with high cardiorespiratory fitness (HIGH CRF) have lower waist circumference (WC), sum of trunk skinfolds (SUM2) and sum of five skinfolds (SUM5), by comparison to those with LOW CRF. DESIGN: Observational cohort study. SETTING: 1981 Canada Fitness Survey (CFS). PARTICIPANTS: A total of 3719 men and 3854 women 20-59 y of age from the 1981 CFS for whom anthropometric and physical fitness measures were available. MAIN OUTCOME MEASURES: BMI, waist circumference, skinfolds, cardiorespiratory fitness. RESULTS: For a given BMI, men and women in the HIGH CRF group had lower WC, SUM2 and SUM5 by comparison to those with LOW CRF (P<0.001). A significant main effect for CRF group remained for SUM2 in both men (P=0.0001) and women (P=0.007), after statistical control for the effects of extremity fat. In women, but not men, there were significant interaction effects for WC (P=0.0023) and SUM5 (P=0.01), indicating that the difference between the LOW CRF and HIGH CRF groups increased with increasing BMI. CONCLUSIONS: HIGH CRF is associated with lower levels of total and abdominal obesity for a given BMI by comparison to those with LOW CRF, independent of gender. This finding suggests a mechanism by which exercise attenuates the health risks attributed to obesity as measured by BMI. It is suggested that the routine measurement of WC and CRF would substantially improve the ability to identify those at health risk. PMID- 12587001 TI - Influence of undersampling on magnetic resonance imaging measurements of intra abdominal adipose tissue. AB - OBJECTIVE: Single-slice magnetic resonance imaging (MRI) and computed tomography (CT) are finding increasing use as methods to estimate visceral fat content in human studies. To assess the validity of this approach, we have compared single- and multi-slice MRI methods for the measurement of intra-abdominal adipose tissue content. MEASUREMENTS: Multi-slice whole-body MR images and single-slices at the level of L2-L3 and L4-L5 were obtained from 59 healthy female volunteers chosen to cover a wide range of body size, and from 17 healthy female volunteers before and after a 6-month exercise intervention. RESULTS: Taking the group as a whole, significant correlation between multi-slice and single-slices was observed (L2 L3: r=0.56; P<0.01; L4-L5: r=0.76; P<0.01). However, the ranking of subjects according to their intra-abdominal fat content was significantly altered by the choice of MRI strategy, especially using L2-L3 methodology. Whole-body (-23.8+/ 20.0%; P<0.01) and single-slice (L2-L3: -31.03+/-35.0%; P<0.01; L4-L5: -22.1+/ 37.24%; p<0.05) MRI methods also detected a significant decrease in intra abdominal fat following the exercise protocol, although the latter techniques gave rise to increased spreading of the data from the mean. These results suggest that the use of single-slice imaging techniques can lead to inconsistencies in the estimation of intra-abdominal fat content, which in turn can have significant effects on data interpretation. CONCLUSION: Single-slice MRI appears to be suitable for assessing changes in intra-abdominal fat content in interventional studies, especially in large cohort of subjects, where each subject can serve as its own control. However, for accurate determination of an individual's intra abdominal fat content, and intersubject comparison, only multi-slice imaging will give precise results. PMID- 12587002 TI - Suppression of insulin secretion is associated with weight loss and altered macronutrient intake and preference in a subset of obese adults. AB - PURPOSE: Hyperinsulinemia is a common feature of many obesity syndromes. We investigated whether suppression of insulin secretion, without dietary or exercise intervention, could promote weight loss and alter food intake and preference in obese adults. METHODS: Suppression of insulin secretion was achieved using octreotide-LAR 40 mg IM q28d for 24 weeks in 44 severely obese adults (89% female, 39% minority). Oral glucose tolerance testing was performed before and after treatment, indices of beta-cell activity (CIRgp), insulin sensitivity (CISI), and clearance (CP/I AUC) were computed, and leptin levels, 3 day food records and carbohydrate-craving measurements were obtained. DEXA evaluations were performed pre- and post-therapy in an evaluable subgroup. RESULTS: For the entire cohort, significant insulin suppression was achieved with simultaneous improvements in insulin sensitivity, weight loss, and body mass index (BMI). Leptin, fat mass, total caloric intake, and carbohydrate craving significantly decreased. When grouped by BMI response, high responders (HR; DeltaBMI<-3 kg/m(2)) and low responders (LR; DeltaBMI between -3 and -0.5) exhibited higher suppression of CIRgp and IAUC than nonresponders (NR; DeltaBMI 0.5). CISI improved and significant declines in leptin and fat mass occurred only in HR and LR. Conversely, both leptin and fat mass increased in NR. Carbohydrate intake was markedly suppressed in HR only, while carbohydrate-craving scores decreased in HR and LR. For the entire cohort, DeltaBMI correlated with DeltaCISI, Deltafat mass, and Deltaleptin. DeltaFat mass also correlated with DeltaIAUC and DeltaCISI. CONCLUSIONS: In a subcohort of obese adults, suppression of insulin secretion was associated with loss of body weight and fat mass and with concomitant modulation of caloric intake and macronutrient preference. PMID- 12587003 TI - Lipolytic and nutritive blood flow response to beta-adrenoceptor stimulation in situ in subcutaneous abdominal adipose tissue in obese men. AB - OBJECTIVE: beta-Adrenoceptor-mediated whole-body lipolysis is impaired in obesity. This study investigated whether local adipocyte beta-adrenergic sensitivity and changes in nutritive blood flow in subcutaneous abdominal adipose tissue contribute to this impaired response. METHODS: Three microdialysis probes were placed in the subcutaneous abdominal adipose tissue of eight obese and nine lean men. Each probe was perfused with either 0.1, 1 and 10 microM isoprenaline; 1, 10 and 100 microM dobutamine or 1, 10 and 100 microM salbutamol, each dose for 45 min. RESULTS: At baseline, interstitial glycerol concentrations and ethanol out/in ratios were comparable between groups. During nonselective beta-, beta(1)- and beta(2)-adrenergic stimulation, interstitial glycerol concentrations increased and ethanol out/in ratios decreased similarly in obese and lean men. CONCLUSION: The lipolytic and nutritive blood flow response to beta(1)- beta(2)- and nonselective beta-adrenergic stimulation in situ is comparable in lean and obese male subjects. The present data suggest that a blunted beta-adrenergic sensitivity of the fat cell and an impaired local nutritive blood flow response do not contribute to the previously reported diminished whole-body beta adrenoceptor-mediated lipolytic response in obese males. PMID- 12587004 TI - Detection of cardiovascular risk factors by indices of obesity obtained from anthropometry and dual-energy X-ray absorptiometry in Japanese individuals. AB - OBJECTIVE: To determine the cut-off points of indices of obesity for detecting hypertension, dyslipidemia and diabetes mellitus in Japanese individuals. DESIGN: Cross-sectional study. SUBJECTS: A total of 2728 Japanese individuals (768 males and 1960 females, aged 20-79 y) who attended the Fukuoka Health Promotion Center, Japan for health check-up. MEASUREMENTS: Body mass index (BMI), waist circumference (WC) and waist-hip ratio (WHR) were measured. Percentage fat mass (%FM), trunk fat mass (FM(trunk)) and trunk fat mass-leg fat mass ratio (FM(trunk)/FM(legs)) were obtained by dual-energy X-ray absorptiometry (DXA). Cardiovascular risk factors were determined by blood pressure, serum lipids, fasting blood glucose and hemoglobin A(1C). RESULTS: The cut-off points of BMI, WC and WHR were around 23.5 kg/m(2), 84 cm and 0.9 for males, and 22.5 kg/m(2), 72 cm and 0.8 for females. The cut-off points of %FM, FM(trunk) and FM(trunk)/FM(legs) were around 24%, 8 kg and 1.6 for males, and 35%, 9 kg and 1.4 for females. WHR and FM(trunk)/FM(legs) most accurately detected the risk factors. CONCLUSIONS: For Japanese individuals, the cut-off points for detecting cardiovascular risk factors are lower than the criteria by the World Health Organization. Indices of fat distribution detected the cardiovascular risk factors more accurately than those of overall adiposity. The accuracy of detecting the risk factors was comparable between the anthropometric indices and indices obtained by DXA. PMID- 12587005 TI - Waist circumference in relation to history of amount and type of alcohol: results from the Copenhagen City Heart Study. AB - OBJECTIVE: To examine the long-term association between the amount and type of alcohol consumed and subsequent high waist circumference. DESIGN: Prospective population study with baseline assessment of alcohol intake, body mass index, smoking habit, physical activity, education, income and deliveries, and after 10 y, examination of waist circumference. SUBJECTS: A sample of 2916 men and 3970 women aged 20-83 y from Copenhagen City Heart Study, Denmark. MEASUREMENTS: A large waist circumference defined as a waist circumference more than 102 cm in men and 88 cm in women. RESULTS: The odds ratios of having a high waist circumference after 10 y showed a linear increase in both men and women, and they were 1.65 (95% confidence interval (CI) 1.07-2.55) in men and 2.16 (0.86-5.14) in women who drank more than 28 beverages per week of total alcohol compared to those who drank one to six beverages per week. Men drinking more than 21 beers per week had odds ratio of having a large waist circumference after 10 y of 1.63 (0.99-2.67) and women drinking more than 14 beers per week had odds ratio of 2.53 (0.92-6.34), compared to men and women who drank no beer. Also for spirits, there was an increase in both men and women. No linear trend was found for wine in either men or women. CONCLUSIONS: Moderate-to-high consumption of alcohol and of beer and spirits was associated with later high waist circumference, whereas moderate-to-high wine consumption may have the opposite effect. PMID- 12587006 TI - The accuracy of long-term recall of past body weight in Japanese adult men. AB - OBJECTIVE: To examine the influences of recall period, current body weight, weight gain and loss, and weight variability on the accuracy of long-term recall of past weight. DESIGN AND SUBJECTS: Comparison of recalled weights around 25 y old with weights measured at age 25y in health checkup records among 2453 Japanese men (age: 34-61 y). MEASUREMENTS: Deviations between reported and measured weights were examined as to the three indexes: actual error (reported measured), percent error (actual error/measured x 100), and absolute value of the percent error. Weight variability was defined as (1) the coefficient of variation of weight (CV) and (2) the root mean square error around the slope of weight on age (RMSE). RESULTS: Recalled weight strongly correlated with measured weight (r=0.849). Correlation coefficients decreased as age or the elapsed time since age 25y increased. Recalled weight (mean=58.3 kg) was slightly greater than measured weight at age 25y (mean=57.0 kg, mean actual error=1.28 kg). Subjects with a current body mass index (BMI) of less than 28.6 kg/m(2) overestimated their past weights, whereas those with BM1 of 28.6 kg/m(2) or over underestimated it. Subjects with weight loss since age 25y underestimated their past weights, whereas those with stable weight or gain overestimated it. There were monotonic increases in the three indexes of deviation across the CV quartile categories. Concerning the effect RMSE, a similar trend was observed. CONCLUSION: These results indicate that past body weights over a long period seem to be recalled with good accuracy. However, it should be kept in mind when using recalled weight in an epidemiologic study that accuracy of recall is influenced by age or elapsed time, current BMI, weight gain and loss, and weight variability. PMID- 12587007 TI - Low body mass index but high percent body fat in Taiwanese subjects: implications of obesity cutoffs. AB - OBJECTIVES: To compare the different correlations of body mass index (BMI) and percent body fat (BF%) with other ethnic groups and to evaluate the appropriateness of Asia-Pacific redefining obesity criteria in the Taiwanese population. The corresponding BF% to BMI cutoffs of overweight and obesity will also be studied. DESIGN: A cross-sectional epidemiological survey. SUBJECTS: A total of 509 male and 570 female Taiwanese subjects aged > or =20 y sampled by the systematic stratified clustering sampling method were analyzed. MEASUREMENTS: BMI was obtained by body weight (kg) divided by squared body height (m(2)). The estimated BMI (BMIe) was deduced from the Caucasian-based four-compartment equation (4C). BF% measured by the methods of dual-energy X-ray absorptiometry (DXA) or Caucasian-based 4C were used and abbreviated as DXA-BF and 4C-BF, respectively. RESULTS: The curvilinear relationship between age and BMI or DXA-BF was established. When compared by age-stratified groups, Taiwanese subjects had a higher BF% (4C-BF) in any given BMI than Caucasians. That is, the level of 4C-BF for BMI> or =25 kg/m(2) in Taiwanese subjects was similar to BMI> or =30 kg/m(2) in Caucasians. The BMIe values of 25 and 30 kg/m(2) were nearly equal to the BMIs of 23.6 and 25.3 kg/m(2) in males, and 22.7 and 24.8 kg/m(2) in females, respectively. The 4C-BF of 25% was nearly equal to a BMI of 26.2 kg/m(2) in males, and 35% was equal to a BMI of 24.4 kg/m(2) in females. Consequently, the DXA-BF cutoffs for BMIs of 23 and 25 kg/m(2) were compatible to 23 and 25% in males, and 35 and 38% in females, respectively. CONCLUSION: It was demonstrated that Taiwanese subjects had a relatively lower BMI but a higher BF% than Caucasians. In general, the newly proposed Asia-Pacific BMI cutoffs for overweight (> or =23 kg/m(2)) and obesity (> or =25 kg/m(2)) may be acceptable to both male and female Taiwanese subjects. The corresponding BF% (DXA-BF) cutoffs for obesity would be 25% in male and 38% in female Taiwanese subjects, respectively. PMID- 12587008 TI - Lower cognitive function in the presence of obesity and hypertension: the Framingham heart study. AB - OBJECTIVE: To determine the independent effects of obesity and hypertension on cognitive functioning. METHODS: Using a prospective design, male (n=551) and female (n=872) participants of the Framingham Heart Study were classified by presence or absence of obesity and hypertension based on data collected over an 18-y surveillance period. All subjects were free from dementia, stroke, and clinically diagnosed cardiovascular disease up to the time of cognitive testing. Statistical models were adjusted for age, education, occupation, cigarette smoking, alcohol consumption, total cholesterol, and a diagnosis of type II diabetes. Body mass index status (nonobese or obese) and blood pressure status (normotensive or hypertensive) were then related to cognitive performance (learning, memory, executive functioning, and abstract reasoning) on tests administered 4-6 y later. RESULTS: Adverse effects of obesity and hypertension on cognitive performance were observed for men only. Obese and hypertensive men performed more poorly than men classified as either obese or hypertensive, and the best performance was observed in nonobese, normotensive men. CONCLUSIONS: The adverse effects of obesity and hypertension in men are independent and cumulative with respect to cognitive deficit. PMID- 12587009 TI - Weighing the care: patients' perceptions of physician care as a function of gender and weight. AB - OBJECTIVE: To examine patients' reports of the level of care that they receive from their physicians, and determine the influence of weight and gender in these reports. DESIGN: : In a four-cell design, male and female, overweight and nonoverweight patients reported on the medical care that they received immediately following their appointment. SUBJECTS: A total of 125 patients affiliated to one of four large clinics in the Texas Medical Center of Houston completed this study. MEASUREMENTS: Patients reported the positivity of the care that they received, the time that physicians spent with them, and the extent to which physicians discussed weight-related topics with them. RESULTS: Overweight patients, as a whole, did not report poorer levels of care than did their thinner counterparts. Rather, the weight and gender of the patient significantly interacted across each of the measures to reveal some divergence between male and female patients' weight-based experiences. When significant differences in reported perceptions emerged, overweight men reported deficits in care relative to average weight men (eg, physicians spent less time), whereas overweight women reported enhanced care relative to average weight women (eg, better levels of care, more topics discussed). CONCLUSION: Based on patients' reports, this study reveals that physician care may not be as influenced by patient weight as previously thought. Yet, there is a discernable impact of patients' weight on physician behavior. Overweight men, who may comprise the most at-risk population, indicate that less time is spent with them than that indicated by average weight men. While this may be alarming, overweight women do not report reductions in care. We propose that not only might physicians respond to them differently, but overweight female patients may also be engaging in denial strategies or compensatory behaviors that assure them of quality care. PMID- 12587010 TI - The agouti-related protein and body fatness in humans. AB - OBJECTIVE: The objective of this study was to examine the impact of a single nucleotide polymorphism (SNP) (-38C>T) in the promoter of the human agouti related protein (hAgRP) gene on promoter affinity for transcription factors (TFs) and its possible association with body composition phenotypes. DESIGN: Electrophoretic mobility shift assays for the functional studies and association analyses for the population studies. SUBJECTS AND METHODS: Nuclear extracts were isolated from the mouse hypothalamus cell line GT1-7 and subjected to binding assays using oligonucleotide probes corresponding to the -38C>T region and an antibody for the E12/E47 TFs. Individuals (n = 259) from the HERITAGE Family Study were genotyped for the -38C>T SNP and used in the association studies. RESULTS: Electrophoretic mobility shift and supershift assays confirmed binding of the E12/E47 TF to the -38C>T site in a genotype-dependent manner. The T allele was found exclusively in the black subjects while the genotype with the higher binding affinity, CC, was significantly associated with high BMI, fat mass, and percent body fat in the black subjects of the HERITAGE Family Study. CONCLUSIONS: The E12/E47 TF could play a role in the regulation of hAgRP expression while the population studies suggest that the TT genotype of the -38C>T SNP could play a protective role against the development of obesity in the black population of the HERITAGE Family Study. PMID- 12587012 TI - NTP Toxicology and Carcinogenesis Studies of Nickel Sulfate Hexahydrate (CAS No. 10101-97-0) in F344 Rats and B6C3F1 Mice (Inhalation Studies). AB - Nickel sulfate hexahydrate is used in nickel plating, as a mordant in dyeing and printing textiles, as a blackening agent for zinc and brass, and in the manufacture of organic nickel salts. Nickel sulfate hexahydrate was nominated by the National Cancer Institute to the NTP as part of a class study of nickel compounds for which there was little information on the toxic and carcinogenic effects of inhalation exposure. Male and female F344/N rats and B6C3F1 mice were exposed to nickel sulfate hexahydrate (greater than 98% pure) by inhalation for 16 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in L5178Y mouse lymphoma cells. 16-DAY STUDY IN RATS: Groups of five male and five female F344/N rats were exposed to 0, 3.5, 7, 15, 30, or 60 mg nickel sulfate hexahydrate/m(3) (equivalent to 0, 0.7, 1.4, 3.1, 6.1, or 12.2 mg nickel/m(3)). Rats were exposed on weekdays only, for a total of 12 exposure days during a 16 day period. Additional groups of four or five male and female F344/N rats were exposed to 0, 3.5, 15, or 30 mg nickel sulfate hexahydrate/m(3)for tissue burden studies. In the core study, two 60 mg/m(3) males, one 30 mg/m(3) female, and all 60 mg/m(3)females died before the end of the study. Final mean body weights of all exposed groups of males and females were significantly lower than those of the controls, as were mean body weight gains of male rats. Clinical findings included increased rates of respiration and reduced activity levels in rats in all exposure groups, except those exposed to 3.5 mg/m(3). Absolute lung weights of 60 mg/m(3) males and of all exposed groups of females were significantly greater than those of the controls, as were the relative lung weights of all exposed groups of males and females. Inflammation (including degeneration and necrosis of the bronchiolar epithelium) occurred in the lungs of all exposed groups of males and females. Atrophy of the olfactory epithelium occurred in the nasal passages of all exposed groups of males (except 60 mg/m(3)) and in 15, 30, and 60 mg/m(3) females. Lymphoid hyperplasia in the bronchial or mediastinal lymph nodes was observed in 30 mg/m(3) males and in 60 mg/m(3) males and females. The concentration of nickel in the lungs of all exposed groups of males and females was greater than in control animals. 16-DAY STUDY IN MICE: Groups of five male and five female B6C3F1 mice were exposed to 0, 3.5, 7, 15, 30, or 60 mg nickel sulfate hexahydrate/m(3). Mice were exposed on weekdays only, for a total of 12 exposure days during a 16-day period. Additional groups of five male and five female B6C3F1 mice were exposed to 0 or 3.5 mg nickel sulfate hexahydrate/m(3)for tissue burden studies. All core study mice exposed to 7 mg/m(3) or greater died before the end of the study; all control and 3.5 mg/m(3)mice survived to the end of the study. Final mean body weights and weight gains of 7, 15, 30, and 60 mg/m(3)males and females were significantly less than those of the controls, and clinical findings in these groups included emaciation, lethargy, and rapid respiration rates. Absolute and relative lung weights of male and female mice exposed to 7 mg/m(3) or greater were significantly greater than those of the controls. Only tissues from mice exposed to 0, 3.5, or 7 mg/m(3) were examined histopathologically. Inflammation occurred in the lungs of 3.5 and 7 mg/m(3) males and females; necrosis of the alveolar and bronchiolar epithelium was a component of the inflammation in 7 mg/m(3)males and females. In addition, atrophy of the olfactory epithelium of the nasal passages was observed in 3.5 mg/m(3) males and females. Nickel concentrations in the lungs of mice exposed to 3.5 mg/m(3) were greater than those in the controls. 13-WEEK STUDY IN RATS: Groups of ten male and ten female F344/N rats were exposed to 0, 0.12, 0.25, 0.5, 1, or 2 mg nickel sulfate hexahydrate (equivalent to 0, 0.03, 0.06, 0.11, 0.22, or 0.44 mg nickel/m(3)), 5 days per week for 13 weeks. Additional groups of six male and six female F344/N rats were exposed to 0, 0.12, 0.5, or 2 mg nic mg nickel sulfate hexahydrate/m(3)for tissue burden studies. In the core study, one 2 mg/m(3)male rat died before the end of the study; all other males and all females survived until the end of the study. Final mean body weights and body weight gains of all exposed groups were similar to those of the controls. There were no significant clinical findings noted during the study. Exposure-related increases in neutrophil and lymphocyte numbers occurred and were most pronounced in female rats. With the exception of 0.12 mg/m(3)rats, absolute and relative lung weights of all exposed groups were generally significantly greater than those of the controls. Exposure-related increases in the incidence and severity of inflammatory lesions (alveolar macrophages, chronic inflammation, and interstitial infiltration) occurred in the lungs of all exposed groups of males and females. Lymphoid hyperplasia of the bronchial and/or mediastinal lymph nodes occurred in males exposed to 0.5 mg/m(3)or greater. Atrophy of the olfactory epithelium occurred in males and females exposed to 0.5, 1, and 2 mg/m(3)and in 0.25 mg/m(3)females. The concentration of nickel in the lungs of 0.5 and 2 mg/m(3) rats was greater than that in the lungs of control animals at 4, 9, and 13 weeks for males and at 13 weeks for females. 13-WEEK STUDY IN MICE: Groups of ten male and ten female B6C3F1 mice were exposed to 0, 0.12, 0.25, 0.5, 1, or 2 mg nickel sulfate hexahydrate, 5 days per week for 13 weeks. Additional groups of up to five or six male and female B6C3F1 mice were exposed to 0, 0.12, 0.5, or 2 mg nickel sulfate hexahydrate/m(3)for tissue burden studies. In the core study, four control males, three control females, and one 0.12 mg/m(3)male died before the end of the study; the deaths were not considered to be chemical related, and all other mice survived to the end of the study. The final mean body weights and body weight gains of all exposed groups were similar to those of the controls. There were no chemical-related clinical findings. Hematology changes similar to those reported in female rats occurred in female mice, but the mice were minimally affected. The absolute and relative lung weights of 1 mg/m(3)males and 2 mg/m(3)males and females were significantly greater than those of the controls. Increased numbers of alveolar macrophages occurred in all males and females exposed to 0.5 mg/m(3)or greater. Chronic active inflammation and fibrosis occurred in 1 and 2 mg/m(3)males and females. Lymphoid hyperplasia of the bronchial lymph node and atrophy of the olfactory epithelium in the nasal passages were observed in 2 mg/m(3)males and females. Nickel concentration in the lung of 2 mg/m(3)females was significantly greater than in control animals. 2 YEAR STUDY IN RATS: Groups of 63 to 65 male and 63 to 64 female rats were exposed to nickel sulfate hexahydrate by inhalation at concentrations of 0, 0.12, 0.25, or 0.5 mg/m(3) (equivalent to 0, 0.03, 0.06, or 0.11 mg nickel/m(3)). Animals were exposed for 6 hours plus T90 (8 minutes) 5 days per week for 104 weeks. Five male and five female rats from each group were evaluated at 7 months for histopathology; an additional seven males and seven females from each group were evaluated at 7 months for nickel tissue burden in the lung and kidney; and five males and five females from each group were evaluated at 15 months for alterations in hematology, nickel tissue burden in the lung and kidney, and histopathology. Survival, Body Weights, Clinical Findings, and Hematology The survival rates of all exposed groups of males and females were similar to those of the controls. Mean body weights of 0.5 mg/m(3)female rats were slightly lower (6% to 9%) than those of the controls throughout the second year of the study; final mean body weights of all exposed groups of males and 0.12 and 0.25 mg/m(3)females were similar to those of the controls. There were no clinical findings or hematology differences that were considered to be related to nickel sulfate hexahydrate administration. Pathology Findings No exposure-related neoplasms occurred in male or female rats exposed by inhalation to nickel sulfate hexahydrate for 2 years. Increased incidences of inflammatory lung lesions were generally observed in all exposed groups of male and female rats at the end of the study. The incidences of chronic active inflammation, macrophage hyperplasia, alveolar proteinosis, and fibrosis were markedly increased in male and female rats exposed to 0.25 or 0.5 mg/m(3). Increased incidences of lymphoid hyperplasia in the bronchial lymph nodes occurred in 0.5 mg/m(3)male and female rats at the end of the 2-year study. The incidences of atrophy of the olfactory epithelium in 0.5 mg/m(3)males and females were significantly greater than those in controls at the end of the study. Tissue Burden Analyses Lung nickel burdens in exposed male and female rats were greater than those in the controls at the 7- and 15-month interim evaluations, and lung nickel burdens values increased with increasing exposure concentration. 2-YEAR STUDY IN MICE: Groups of 80 male and 80 female mice were exposed to nickel sulfate hexahydrate by inhalation at concentrations of 0, 0.25, 0.5, or 1 mg/m(3) (equivalent to 0, 0.06, 0.11, or 0.22 mg nickel/m(3)). Animals were exposed for 6 hours plus T90 (8 minutes) 5 days per week for 104 weeks. Five male and five female mice from each group were evaluated at 7 months for histopathology; five males and five females from each group were evaluated at 7 months for nickel tissue burden in the lung and kidney; five males and five females from each group were evaluated at 15 months for alterations in hematology and histopathology; and five males and five females from each group were evaluated at 15 months for nickel tissue burden in the lung and kidney. Survival, Body Weights, Clinical Findings, and Hematology The survival rates of all exposed groups of males and females were similar to those of the controls. The mean body weights of 1 mg/m(3)males and of all exposed groups of females were lower than those of the controls during the second year of the study. There were no clinical findings or hematology differences considered to be related to chemical exposure. Pathology Findings Inflammatory lesions of the lung generally occurred in all exposed groups of male and female mice at the end of the 2-year study. These lesions included macrophage hyperplasia, chronic active inflammation, bronchialization (alveolar epithelial hyperplasia), alveolar proteinosis, and infiltrating cells in the interstitium. Incidences of macrophage hyperplasia and/or lymphoid hyperplasia occurred in the bronchial lymph nodes of most of the 1 mg/m(3)males and females and in some 0.5 mg/m(3)females at the end of the 2-year study. Atrophy of the olfactory epithelium was observed in 0.5 and 1 mg/m(3)males and in all exposed groups of females at the end of the 2-year study. Tissue Burden Analyses At the 7- and 15-month interim evaluations, lung nickel burden parameters measured in control and exposed groups were below the limit of detection. Absolute lung weights of 0.5 and 1 mg/m(3)lung burden study females were significantly greater than those of the controls at 15 months. GENETIC TOXICOLOGY: Nickel sulfate hexahydrate (500 to 800 g/mL) was tested for induction of trifluorothymidine resistance in L5178Y mouse lymphoma cells. A positive response was observed in the absence of S9. The test was not performed with S9. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was no evidence of carcinogenic activity of nickel sulfate hexahydrate in male or female F344/N rats exposed to 0.12, 0.25, or 0.5 mg/m(3) (0.03, 0.06, or 0.11 mg nickel/m(3)). There was no evidence of carcinogenic activity of nickel sulfate hexahydrate in male or female B6C3F1 mice exposed to 0.25, 0.5, or 1 mg/ m3 (0.06, 0.11, or 0.22 mg nickel/m(3)). Exposure of rats to nickel sulfate hexahydrate by inhalation for 2 years resulted in increased incidences of chronic active inflammation, macrophage hyperplasia, alveolar proteinosis, and fibrosis of the lung; lymphoid hyperplasia of the bronchial lymph node; and atrophy of the olfactory epithelium. Exposure of mice to nickel sulfate hexahydrate by inhalation for 2 years resulted in increased incidences of chronic active inflammation, bronchialization (alveolar epithelial hyperplasia), macrophage hyperplasia, interstitial infiltration, and alveolar proteinosis of the lung; lymphoid and macrophage hyperplasia of the bronchial lymph node; and atrophy of the olfactory epithelium. Synonyms: Blue salt; hexahydrate, nickel (2+) salt; nickel monosulfate hexahydrate; nickel (2+) sulfate hexahydrate; nickel (II) sulfate hexahydrate; nickel sulphate hexahydrate; nickelous sulfate hexahydrate; nickelous sulphate hexahydrate; single nickel salt, sulfuric acid PMID- 12587011 TI - Body mass index and the risk of mortality in type II diabetic patients from Verona. AB - OBJECTIVE: The relation between body weight and mortality in type II diabetic patients has not been fully elucidated. The aim of the present study was to evaluate the impact of body weight on mortality in a well-characterized type II diabetic cohort. RESEARCH DESIGN AND METHODS: We examined a cohort of 3398 type II diabetic patients, alive on December 1986 and followed up for 10 years, to assess mortality and its causes and to investigate the relationship between body mass index (BMI) and mortality from all and specific causes. For this purpose, survival in the different quartiles of BMI was evaluated by a Cox model, controlling for sex, age, treatment, smoking, duration of diabetes, hypertension, and fasting plasma glucose. The Cox model was applied either excluding (model 1) or including (model 2) the last three variables. RESULTS: During the 10 ys of follow-up, 1212 deaths (639 women, 573 men) occurred in the cohort under study. Since the interaction between BMI and age was statistically significant (P = 0.002), survival was studied separately in people aged <65 and > or =65 y (median age of the cohort = 65.9 y). Under 65 y, a significantly higher all-cause mortality was observed in obese patients, that is, in the IV quartile (BMI> or =30.9 kg/m(2); RR = 1.74; CI 95% = 1.26-2.40), in model 1. The inclusion of hypertension, duration of diabetes, and fasting plasma glucose in the model (model 2) slightly decreased the relative risk (RR = 1.52; CI 95% = 1.10-2.11). After 65 y, higher body weight was associated with a better outcome, especially in patients belonging to the IV quartile of BMI (RR = 0.74; CI 95% = 0.62-0.90). CONCLUSIONS: In older type II diabetic patients, a moderate excess weight predicts a better survival, while obesity is a negative prognostic factor in patients younger than 65 y. In the latter patients, the effect of obesity on mortality seems to be partly mediated by hypertension, duration of diabetes, and fasting plasma glucose. PMID- 12587013 TI - NTP Toxicology and Carcinogenesis Studies of D&C Yellow No. 11 (CAS No. 8003-22 3) in F344/N Rats (Feed Studies). AB - D&C Yellow No. 11 is used to color topical drug preparations and cosmetics. It is also used in spirit lacquers, polystyrenes, polycarbonates, polyamides, acrylic resins, colored smokes, and hydrocarbon solvents. D&C Yellow No. 11 was nominated to the NTP for toxicity and carcinogenesis studies as part of a larger regulatory effort mandated by Congress and undertaken by the Food and Drug Administration to determine the safety of a number of provisionally listed dyes. D&C Yellow No. 11 is currently regulated for external use. The recommendation to study D&C Yellow No. 11 by dietary exposure was based on the fact that it is a contaminant of D&C Yellow No. 10, a candidate for permanent listing as a chemical for which there is a potential for ingestion. First-generation (F(0)) male and female F344/N rats were given D&C Yellow No. 11 (approximately 99% pure) in feed for up to 19 weeks and then mated, and exposure of second-generation (F(1)) males and females began in utero and continued for 2 years after weaning at 28 days of age. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and mouse peripheral blood. REPRODUCTIVE TOXICITY STUDY: Groups of 60 male and 60 female F(0) rats were given 0, 500, 1,700, or 5,000 ppm D&C Yellow No. 11 in feed for up to 19 weeks, which resulted in average daily doses of 35, 120, or 350 mg D&C Yellow No. 11/kg body weight to males and 35, 120, or 370 mg/kg to females. All F(0) males and females survived until the end of the study. Prior to cohabitation, mean body weight gains of males given 500, 1,700, or 5,000 ppm and of females given 5,000 ppm were significantly lower than those of the controls. The mean body weight gains of exposed females during gestation and lactation were generally similar to those of the controls. Feed consumption by exposed groups of rats was generally similar to that by the control groups prior to cohabitation. The duration of gestation, the average litter size, the number of live pups on days 4 (precull) and 21, and the percentage of male pups for each exposure group were similar to those of the controls. The mean body weights of exposed litters were significantly less than those of the control litters on days 14 and 21; this effect was considered to be related to D&C Yellow No. 11 exposure. 2-YEAR STUDY: Groups of 60 male and 60 female F(1) rats were given 0, 500, 1,700, or 5,000 ppm D&C Yellow No. 11 in feed for 105 (males) or 106 (females) weeks after weaning (day 28); 6 to 10 rats per group were evaluated at 12 months. These exposure concentrations resulted in average daily doses of approximately 25, 85, or 250 mg D&C Yellow No. 11/kg body weight to males and 25, 100, or 280 mg/kg to females. Survival, Body Weights, Feed Consumption, and Clinical Findings: Survival of males given 1,700 or 5,000 ppm was significantly less than that of the controls, and survival of 1,700 ppm females was significantly greater than that of the controls. Mean body weights of 1,700 and 5,000 ppm males and females were generally lower than those of the controls throughout the study. Feed consumption by exposed groups was similar to that by the controls. Chemical-related clinical findings included yellow discoloration of the entire body in all exposed males and females from day 1 and head swelling and edema in 1,700 and 5,000 ppm males. One 1,700 ppm and five 5,000 ppm males were moribund and were killed between weeks 49 and 81; these deaths were attributed to extensive edema. Hematology: A few minimal hematology changes occurred in male rats at the 12-month interim evaluation. There was evidence of minimal anemia in exposed males; this anemia was characterized by decreased hematocrit values, hemoglobin concentrations, and erythrocyte counts. The minimal anemia was characterized as normocytic, normochromic, and nonresponsive. There were no biologically or statistically significant differences in hematology parameters between control and exposed females. Pathology Findings: Absolute and relative liver weights of all exposed groups of md relative liver weights of all exposed groups of males and females were significantly greater than those of the controls at 12 months. At 2 years, the incidences of hepatocellular adenoma in 5,000 ppm males and of hepatocellular adenoma or carcinoma (combined) in 5,000 ppm females were significantly greater than those in the controls. At 12 months, the incidences of clear cell foci in 1,700 and 5,000 ppm females were significantly greater than that in the controls. At 2 years, the incidences of mixed cell foci in exposed males and of clear cell foci in exposed males (except 500 ppm) and females were significantly greater than those in the controls. Incidences of cytologic alterations (basophilia and granularity) of hepatocytes, and pigmentation in bile duct epithelium, hepatocytes, and Kupffer cells in exposed males and females were greater than those in the controls at both 12 months and 2 years. Renal tubule adenomas were observed in two 5,000 ppm males, and one renal tubule carcinoma was observed in a 1,700 ppm male. During an extended evaluation, renal tubule adenomas were observed in two additional 5,000 ppm males, four 1,700 ppm males, and two 500 ppm males. Renal tubule hyperplasia was observed in exposed groups of males but not in controls, and the incidences in 1,700 ppm males from both standard and extended evaluations were significantly greater than those in the controls. Necrosis and regeneration of the renal tubule epithelium were observed in all control and exposed male rats and in most female rats at 12 months and 2 years. The severity of nephropathy in exposed males and females was significantly greater than that in the controls. In exposed males and 1,700 ppm females at 2 years, the incidences of hyperplasia of the transitional epithelium in the kidney, which commonly accompanies advanced nephropathy, were greater than those of the controls, and the severity of this lesion in exposed males and females was greater than that in the controls. The incidences of renal tubule pigmentation in all exposed groups of males and females at 12 months and 2 years were significantly greater than those in the controls. Squamous cell carcinomas of the tongue were observed in one 500 ppm male at 12 months and one 5,000 ppm female at 2 years, and one squamous cell carcinoma of the oral mucosa was observed in each group of exposed males and in one 5,000 ppm female at 2 years. At 2 years, squamous cell papillomas were observed in the oral cavity (oral mucosa or tongue) of one control, one 500 ppm, two 1,700 ppm, and four 5,000 ppm males; this lesion was also observed in one control and one 500 ppm female. GENETIC TOXICOLOGY: Results of mutagenicity tests with D&C Yellow No. 11 in Salmonella typhimurium were equivocal in one study, based on responses observed in strain TA100 with induced rat liver S9, and weakly positive in a second study, based on responses observed in strains TA98 and TA100 with induced rat or hamster liver S9. D&C Yellow No. 11 induced sister chromatid exchanges and chromosomal aberrations in cultured Chinese hamster ovary cells, with and without S9. No increase in the frequency of micronucleated normochromatic erythrocytes was observed in peripheral blood samples from male and female B6C3F(1) mice administered D&C Yellow No. 11 in feed for 13 weeks. CONCLUSIONS: Under the conditions of this perinatal exposure followed by a 2-year dosed feed study, there was some evidence of carcinogenic activity of D&C Yellow No. 11 in male F344/N rats based on increased incidences of hepatocellular adenoma, renal tubule neoplasms, and squamous cell neoplasms of the oral cavity. There was some evidence of carcinogenic activity in female F344/N rats based on increased inci dences of hepatocellular neoplasms. Incidences of uncommon squamous cell carcinoma of the oral cavity in females may have been related to chemical treatment. Exposure of rats to D&C Yellow No. 11 in feed for 2 years resulted in increased incidences of nonneoplastic liver lesions including clear cell foci, increased basophilia and granularity in the cytoplasm of hepatocytes, and bile duct, hepatocyte, and Kupffer cell pigmentation in males and females and mixed cell foci in males. In the kidney, there were increased incidences of renal tubule pigmentation and transitional epithelial hyperplasia in males and females and renal tubule hyperplasia in males. The severity of nephropathy was increased in exposed males and females. Synonyms: 2-(2-Quinolinyl)-1H-indene-1,3-(2H)-dione; 2-(2-quinolyl) 1,3-indandione Trade names: Arlosol Yellow S, Chinoline Yellow D (soluble in spirits), Chinoline Yellow ZSS, C.I. 47000, C.I. Solvent Yellow 33, Nitro Fast Yellow SL, Oil Yellow SIS, Petrol Yellow C, Quinoline Yellow A Spirit Soluble, Quinoline Yellow Base, Quinoline Yellow Spirit Soluble, Quinoline Yellow SS, Solvent Yellow 33, Waxoline Yellow T PMID- 12587014 TI - NTP Toxicology and Carcinogenesis Studies of Molybdenum Trioxide (CAS No. 1313-27 5) in F344 Rats and B6C3F1 Mice (Inhalation Studies). AB - Molybdenum is an essential element for the function of nitrogenase in plants and as a cofactor for enzymes including xanthine oxidoreductase, aldehyde oxidase, and sulfide oxidase in animals. Molybdenum trioxide is used primarily as an additive to steel and corrosion-resistant alloys. It is also used as a chemical intermediate for molybdenum products; an industrial catalyst; a pigment; a crop nutrient; components of glass, ceramics, and enamels; a flame retardant for polyester and polyvinyl chloride resins; and a reagent in chemical analyses. Molybdenum trioxide was nominated by the NCI for toxicity and carcinogenicity studies as a representative inorganic molybdenum compound. The production of molybdenum trioxide is the largest of all the molybdenum compounds examined. Male and female F344/N rats and B6C3F1 mice were exposed to molybdenum trioxide (approximately 99% pure) by inhalation for 14 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and cultured Chinese hamster ovary cells. 14-DAY STUDY IN RATS: Groups of five male and five female F344/N rats were exposed to 0, 3, 10, 30, 100, or 300 mg molybdenum trioxide/m(3). Rats were exposed for 6 hours per day, 5 days per week, for a total of 10 exposure days during a 14-day period. All rats survived to the end of the study. The final mean body weights of male rats exposed to 100 mg/m(3) and male and female rats exposed to 300 mg/m(3) were significantly lower than those of the control groups. Male rats exposed to 300 mg/m(3) lost weight during the study. There were no clinical findings related to exposure to molybdenum trioxide. No chemical-related lesions were observed. 14-DAY STUDY IN MICE: Groups of five male and five female B6C3F1 mice were exposed to 0, 3, 10, 30, 100, or 300 mg molybdenum trioxide/m(3). Mice were exposed 6 hours per day, 5 days per week, for a total of 10 exposure days during a 14-day period. All mice survived to the end of the study. Final mean body weights of male and female mice exposed to 300 mg/m(3) were significantly lower than those of the control groups. Male mice exposed to 300 mg/m(3) lost weight during the study. There were no clinical findings related to exposure to molybdenum trioxide. No chemical-related lesions were observed. 13-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were exposed to molybdenum trioxide by inhalation at concentrations of 0, 1, 3, 10, 30, or 100 mg/m(3) for 6.5 hours per day, 5 days per week, for 13 weeks. All rats survived to the end of the study. The final mean body weights of exposed rats were similar to those of the control groups. No clinical findings related to molybdenum trioxide exposure were observed. There were no significant chemical related differences in absolute or relative organ weights, hematology or clinical chemistry parameters, sperm counts or motility, or liver copper concentrations between control and exposed rats. No chemical-related lesions were observed. 13 WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were exposed to molybdenum trioxide by inhalation at concentrations of 0, 1, 3, 10, 30, or 100 mg/m(3) for 6.5 hours per day, 5 days per week, for 13 weeks. All mice survived to the end of the study. The final mean body weights of exposed mice were similar to those of the control groups. There were no chemical-related clinical findings. There were no significant differences in absolute or relative organ weights or sperm counts or motility between control and exposed mice. There were significant increases in liver copper concentrations in female mice exposed to 30 mg/m(3) and in male and female mice exposed to 100 mg/m(3) compared to those of the control groups. No chemical-related lesions were observed. 2-YEAR STUDIES IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to molybdenum trioxide by inhalation at concentrations of 0, 10, 30, or 100 mg/m(3). Rats were exposed for 6 hours per day, 5 days per week, for 106 weeks. Survival, Body Weights, and Special Studies: Survival rates of exposed maleed male and female rats were similar to those of the control groups. Mean body weights of exposed groups of male and female rats were similar to those of the control groups throughout the study. There was a significant exposure-dependent increase in blood molybdenum concentration in exposed rats. Blood concentrations of molybdenum in exposed male rats were greater than those in exposed female rats. There were no toxicologically significant differences in bone density or curvature between control and exposed rats. Pathology Findings: The incidences of alveolar/bronchiolar adenoma or carcinoma (combined) were increased in male rats with a marginally significant positive trend. No increase in the incidences of lung neoplasms occurred in female rats. Incidences of chronic alveolar inflammation in male and female rats exposed to 30 or 100 mg/m(3) were significantly greater than those in the control groups. No nasal or laryngeal neoplasms were attributed to exposure to molybdenum trioxide. Incidences of hyaline degeneration in the nasal respiratory epithelium in 30 and 100 mg/m(3) males and in all exposed groups of females were significantly greater than those in the control groups. The incidences of hyaline degeneration in the nasal olfactory epithelium of all exposed groups of females were significantly greater than that in the control group. In the larynx, incidences of squamous metaplasia of the epithelium lining the base of the epiglottis in all exposed groups of male and female rats were significantly greater than those in the control groups and increased with increasing exposure concentration. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were exposed to molybdenum trioxide by inhalation at concentrations of 0, 10, 30, or 100 mg/m(3). Mice were exposed for 6 hours per day, 5 days per week, for 105 weeks. Survival, Body Weights, and Special Studies: The survival rate of male mice exposed to 30 mg/m(3) was marginally lower than that of the control group; survival rates of 10 and 100 mg/m(3) males and of all exposed groups of females were similar to those of the control groups. Mean body weights of exposed male mice were generally similar to those of the control group throughout the study. Mean body weights of exposed female mice were generally greater than those of the control group from week 11 until the end of the study. There was a significant exposure-dependent increase in blood molybdenum concentration in exposed mice. There were no toxicologically significant differences in bone density or curvature between control and exposed mice. Pathology Findings: The incidences of alveolar/bronchiolar carcinoma in all exposed groups of males were significantly greater than that in the control group. Incidences of alveolar/bronchiolar adenoma in females in the 30 and 100 mg/m(3) groups were significantly greater than that in the control group. Incidences of alveolar/bronchiolar adenoma or carcinoma (combined) in 10 and 30 mg/m(3) males and in 100 mg/m(3) females were significantly greater than those in the control groups and exceeded the historical control ranges for 2-year NTP inhalation studies. Incidences of metaplasia of the alveolar epithelium of minimal severity in the centriacinar region of the lung were significantly increased in all exposed groups of mice. The incidences of histiocyte cellular infiltration in all exposed groups of males were significantly greater than that in the control group. Incidences of hyaline degeneration of the respiratory epithelium of the nasal cavity in 100 mg/m(3) males and females and hyaline degeneration of the olfactory epithelium of the nasal cavity in 100 mg/m(3) females were significantly greater than those in the control groups. The incidences of squamous metaplasia of the epithelium lining the base of the epiglottis were significantly increased in all exposed groups of males and females. In both male and female mice, the incidences of hyperplasia of the laryngeal epithelium in level II of the larynx increased with increasing exposure concentration. The increase was statistically significant only in mice exposed to 100 mg/m(3) with 82% of male and 70% of female mice affected. GENETIC TOXICOLOGY: Molybdenum trioxide was not mutagenic in any of five strains of Salmonella typhimurium, and it did not induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells in vitro. All tests were conducted with and without S9 metabolic activation enzymes. CONCLUSIONS: Under the conditions of these 2-year inhalation studies, there was equivocal evidence of carcinogenic activity of molybdenum trioxide in male F344/N rats based on a marginally significant positive trend of alveolar/bronchiolar adenoma or carcinoma (combined). There was no evidence of carcinogenic activity of molybdenum trioxide in female F344/N rats exposed to 10, 30, or 100 mg/m(3). There was some evidence of carcinogenic activity of molybdenum trioxide in male B6C3F1 mice based on increased incidences of alveolar/bronchiolar carcinoma and adenoma or carcinoma (combined). There was some evidence of carcinogenic activity of molybdenum trioxide in female B6C3F1 mice based on increased incidences of alveolar/bronchiolar adenoma and adenoma or carcinoma (combined). Exposure of male and female rats to molybdenum trioxide by inhalation resulted in increased incidences of chronic alveolar inflammation, hyaline degeneration of the respiratory epithelium, hyaline degeneration of the olfactory epithelium (females), and squamous metaplasia of the epiglottis. Exposure of male and female mice to molybdenum trioxide by inhalation resulted in increased incidences of metaplasia of the alveolar epithelium, histiocyte cellular infiltration (males), hyaline degeneration of the respiratory epithelium, hyaline degeneration of the olfactory epithelium (females), squamous metaplasia of the epiglottis, and hyperplasia of the larynx. Synonyms: Molybdic oxide; molybdic trioxide; molybdic anhydride; molybdenum (VI) oxide; molybdenum peroxide; molybdic acid anhydride; molybdenum anhydride; natural molybdite; molybdena PMID- 12587015 TI - NTP Toxicology and Carcinogenesis Studies of Nitromethane (CAS No. 75-52-5) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Nitromethane is used as a rocket and engine fuel; as a synthesis intermediate for agricultural fumigants, biocides, and other products; as a solvent; and as an explosive in mining, oil-well drilling, and seismic exploration. It has been detected in air, in surface and drinking water, and in cigarette smoke. Nitromethane was studied because of the potential for widespread human exposure and because it is structurally related to the carcinogens 2-nitropropane and tetranitromethane. Male and female F344/N rats and B6C3F1 mice received nitromethane (purity 98% or greater) by inhalation for 16 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and peripheral blood erythrocytes of mice. 16-DAY STUDY IN RATS: Groups of five male and five female rats were exposed to 0, 94, 188, 375, 750, or 1,500 ppm nitromethane by inhalation, 6 hours per day, 5 days per week, for 16 days. All rats survived until the end of the study. The mean body weight gain of male rats in the 1,500 ppm group was slightly but significantly less than that of the controls; the final mean body weights and mean body weight gains of exposed females were similar to those of the controls. Clinical findings in all male and female rats in the 1,500 ppm groups included increased preening, rapid breathing, hyperactivity early in the study, and hypoactivity and loss of coordination in the hindlimbs near the end of the study. The relative liver weights of all exposed groups of male rats and the absolute and relative liver weights of females exposed to 375 ppm or greater were significantly greater than those of the controls. Minimal to mild degeneration of the olfactory epithelium was observed in the nose of males and females exposed to 375 ppm or greater. Sciatic nerve degeneration was present in all male and female rats exposed to 375 ppm or greater; rats exposed to 750 or 1,500 ppm also had reduced myelin around sciatic axons. 16-DAY STUDY IN MICE: Groups of five male and five female mice were exposed to 0, 94, 188, 375, 750, or 1,500 ppm nitromethane by inhalation, 6 hours per day, 5 days per week, for 16 days. All mice survived to the end of the study. The final mean body weights and weight gains of exposed males and females were similar to those of the controls. Clinical findings included hypoactivity and tachypnea in male and female mice in the 1,500 ppm groups. Absolute and relative liver weights of male mice in the 750 and 1,500 ppm groups and female mice in all exposed groups and the relative liver weight of males in the 375 ppm group were significantly greater than those of the controls. Degeneration of the olfactory epithelium of the nose was observed microscopically in all males and females exposed to 375 ppm or greater; this lesion was of minimal severity in males and minimal to mild severity in females. 13-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to 0, 94, 188, 375, 750, or 1,500 ppm nitromethane by inhalation, 6 hours per day, 5 days per week, for 13 weeks. All rats survived to the end of the study. The final mean body weight and weight gain of male rats in the 1,500 ppm group were significantly less than those of the controls. Clinical findings included hindlimb paralysis in rats in the 750 and 1,500 ppm groups. Inhalation exposure of rats to nitromethane resulted in an exposure concentration-dependent, microcytic, responsive anemia; anemia was most pronounced in males and females exposed to 375 ppm or greater. The presence of schistocytes, Heinz bodies, and spherocytes and increased mean cell hemoglobin concentration and methemoglobin concentration were evidence that a hemolytic process was occurring; this hemolytic process could have accounted, in part, for the anemia. Thrombocytosis accompanied the anemia and would be consistent with a reactive bone marrow or could have been due to the erroneous inclusion of small erythrocyte fragments as part of the platelet count. On day 23, transient decreases in serum triiodothyronine, thyroxine, and fr and free thyroxine were observed in male rats exposed to 375 ppm or greater and female rats exposed to 750 or 1,500 ppm. There was little or no pituitary response to the thyroid hormone decreases, as evidenced by the lack of significantly increased concentrations of thyroid stimulating hormone in exposed rats. No biologically significant differences in organ weights were observed. The forelimb and hindlimb grip strengths of males in the 1,500 ppm group were significantly less than those of the controls. The hindlimb grip strengths of females in the 750 and 1,500 ppm groups were also significantly less than the control value. Minimal to mild hyperplasia of the bone marrow was observed microscopically in male rats in the 750 and 1,500 ppm groups and in females exposed to 188 ppm or greater. Nasal lesions in exposed males and females included olfactory epithelial degeneration in males and females exposed to 375 ppm or greater and in one female exposed to 188 ppm and respiratory epithelial hyaline droplets and goblet cell hyperplasia in males and females in the 750 and 1,500 ppm groups; the severity of nasal lesions in males and females was minimal to mild. Males and females exposed to 375 ppm or greater had minimal to mild degeneration of the sciatic nerve and the lumbar spinal cord. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were exposed to 0, 94, 188, 375, 750, or 1,500 ppm nitromethane by inhalation, 6 hours per day, 5 days per week, for 13 weeks. All mice survived to the end of the study. The final mean body weights and weight gains of exposed mice were generally similar to those of the controls. There were no treatment-related clinical findings. The absolute right kidney weights of all groups of exposed male mice except the 1,500 ppm group and of females exposed to 188 ppm or greater and the relative right kidney weights of all groups of exposed males and of females in the 750 and 1,500 ppm groups were significantly greater than those of the controls. The absolute liver weight of male mice in the 750 ppm group and the relative liver weights of males exposed to 375 ppm or greater were significantly greater than those of the controls. Olfactory epithelial degeneration and respiratory epithelial hyaline droplets were observed microscopically in all male and female mice exposed to 375 ppm or greater. Degeneration also occurred in females in the 188 ppm group, and hyaline droplets occurred in females in the 94 and 188 ppm groups. The average severity of the nasal lesions ranged from minimal to mild in males. In females, the average severity of olfactory epithelial degeneration ranged from minimal to mild and the severity of respiratory epithelial hyaline droplets ranged from minimal to moderate. All males and nine females in the 1,500 ppm groups also had minimal extramedullary hematopoiesis of the spleen. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed to 0, 94, 188, or 375 ppm nitromethane by inhalation, 6 hours per day, 5 days per week, for 103 weeks. Survival,Body Weights, and Clinical Findings: There were no significant differences in survival rates between exposed and control male or female rats. The mean body weight of females in the 375 ppm group was slightly greater than that of the control group; the mean body weights of exposed males were generally similar to the mean body weight of the controls throughout the study. Clinical findings were consistent with incidences of mammary gland neoplasms in females exposed to 188 or 375 ppm; no hindlimb paralysis, as occurred in rats in the 13-week study, was observed in male or female rats in the 2-year study. Pathology Findings: The incidences of mammary gland fibroadenoma and fibroadenoma, adenoma, or carcinoma (combined) in female rats in the 188 and 375 ppm groups were significantly greater than those in the controls. Additionally, the incidences of mammary gland carcinoma in the 375 ppm group were significantly greater than those in the controls. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed to 0, 188, 375, or 750 ppm nitromethane by inhalation, 6 hours per day, 5 days per week, for 103 weeks. Survival,Body Weights, and ClinicalFindings The survival rate of females in the 750 ppm group was marginally greater than that of the controls. The mean body weights of exposed females were generally slightly greater than the mean body weights of the controls during the study but were generally similar to the mean body weight of the controls at the end of the study. The mean body weights of exposed males were similar to those of the controls throughout the study. Clinical findings included swelling around the eyes and exophthalmos in exposed males and females; these findings were coincident with harderian gland neoplasms. Pathology Findings: The incidences of harderian gland adenoma and adenoma or carcinoma (combined) in exposed mice increased with increasing exposure concentration and were significantly greater in males and females in the 375 and 750 ppm groups than those in the controls. The incidences of harderian gland carcinoma in males and females in the 375 and 750 ppm groups were also slightly greater than those in the controls. Female mice in the 188 and 750 ppm groups had significantly greater incidences of hepatocellular adenoma and hepatocellular adenoma or carcinoma (combined) than the controls. The incidences of liver eosinophilic focus increased with increasing exposure concentration, and the incidences in the 375 and 750 ppm groups were significantly greater than the control incidence. The incidences of alveolar/bronchiolar carcinoma in male mice in the 750 ppm group and female mice in the 375 ppm group were significantly greater than those in the controls. Females in the 750 ppm group also had a significantly greater incidence of alveolar/bronchiolar adenoma or carcinoma (combined) and a slightly greater incidence of alveolar/bronchiolar adenoma than the controls. Females in the 375 ppm group had a significantly greater incidence of cellular infiltration of histiocytes in the lung than the controls. The incidences of degeneration and metaplasia of the olfactory epithelium and hyaline degeneration of the respiratory epithelium were significantly greater in exposed male and female mice than those in the controls. Additionally, males in the 375 and 750 ppm groups had significantly greater incidences of inflammation of the nasolacrimal duct than did the controls. GENETIC TOXICOLOGY: Nitromethane was not mutagenic in any tests performed by the NTP. It did not induce mutations in Salmonella typhimurium, with or without S9 metabolic activation, and no induction of sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells exposed to nitromethane was noted with or without S9. No increase in the frequency of micronucleated erythrocytes was observed in peripheral blood samples of male and female mice at the end of the 13-week inhalation study of nitromethane. CONCLUSIONS: Under the conditions of these 2 year inhalation studies, there was no evidence of carcinogenic activity of nitromethane in male F344/N rats exposed to 94, 188, or 375 ppm. There was clear evidence of carcinogenic activity of nitromethane in female F344/N rats based on increased incidences of mammary gland fibroadenomas and carcinomas. There was clear evidence of carcinogenic activity of nitromethane in male B6C3F1 mice based on increased incidences of harderian gland adenomas and carcinomas. There was clear evidence of carcin ogenic activity in female B6C3F1 mice, based on increased incidences of liver neoplasms (primarily adenomas) and harderian gland adenomas and carcinomas. Increased incidences of alveolar/bronchiolar adenomas and carcinomas in male and female mice exposed to nitromethane were also considered to be related to chemical administration. Exposure to nitromethane by inhalation for 2 years resulted in increased incidences of nasal lesions including degeneration and metaplasia of the olfactory epithelium and degeneration of the respiratory epithelium in male and female mice. Synonym: Nitrocarbol PMID- 12587016 TI - Effect of Dietary Restriction on Toxicology and Carcinogenesis Studies in F344/N Rats and B6C3F1 Mice. AB - Studies were conducted to compare outcomes when four chemicals were evaluated under typical NTP bioassay conditions as well as under protocols employing dietary restriction. Specific experiments were designed to evaluate the effect of diet restriction on the sensitivity of the bioassay toward chemical-induced chronic toxicity and carcinogenicity and to evaluate the effect of weight-matched control groups on the sensitivity of the bioassays. Two chemicals, butyl benzyl phthalate and t-butylhydroquinone, were administered in feed; one chemical, salicylazosulfapyridine, was administered in corn oil by gavage; and one chemical, scopolamine hydrobromide trihydrate, was administered in distilled water by gavage. In each of four protocols, the effects of the chemical were assessed by a comparison between a group exposed to a single dose concentration of the study chemical and a nonexposed control group. F344/N rats and B6C3F1 mice were fed NIH-07 diet either ad libitum or in amounts that restricted mean body weights according to the following design requirements. For the core bioassay, groups of 50 to 60 ad libitum-fed animals were allotted to a control group and three dosed groups for approximately 104 weeks or up to 128 weeks (t butylhydroquinone study). The comparison between the control group and the group receiving the highest dose was used to represent the outcome of the bioassay under ad libitum feeding protocols. In a second comparison, outcomes from the group receiving the highest dose were compared with a weight-matched group of 50 to 60 untreated controls; the weight-matched controls received feed in amounts restricted so that the mean body weight matched the mean body weight of the dosed group. Two additional groups of 48 to 60 animals (one control and one dosed group) were offered feed in amounts that limited the mean body weight of the control group to approximately 85% that of the controls fed ad libitum under the first protocol. Animals assigned to this dietary restriction paradigm were evaluated after 104 weeks or 130 weeks (t-butylhydroquinone). A fourth protocol was em- loyed to evaluate whether an additional period of exposure (up to 1 year) would influence the neoplasm profile of animals fed a restricted diet. Two groups of approximately 50 animals (one control and one dosed group) in the butyl benzyl phthalate, salicylazosulfapyridine, and scopolamine hydrobromide trihydrate studies received restricted diets, as under the third protocol, for 3 years or until survival in either group was reduced to 20%. Butyl benzyl phthalate caused an increased incidence of pancreatic acinar cell neoplasms in ad libitum-fed male rats relative to ad libitum-fed and weight-m atched controls. This change did not occur in rats in the restricted feed protocol after 2 years; however, acinar cell adenomas were observed in three exposed, feed-restricted males at 30 months. Feed restriction is known to influence the incidence of pancreatic acinar cell neoplasms and may have prevented the full expression of this chemical-induced effect. Butyl benzyl phthalate also caused an increased incidence of urinary bladder neoplasms in female rats in the 32-month restricted feed protocol. The incidences of urinary bladder neoplasms were not significantly increased in female rats in any of the 2-year protocols, suggesting that the length of study, and not body weight, was the primary factor in the detection of this carcinogenic response. Salicylazosulfapyridine caused an increased incidence of urinary bladder papillomas in male rats fed ad libitum relative to ad libitum-fed and weight- matched controls. This increase was associated with an increased incidence of urinary bladder calculi; the incidences of urinary bladder concretions, dilatation, and hyperplasia were also increased in dosed males. The incidences of urinary bladder papillomas and calculi were not increased in male rats receiving salicylazosulfapyridine that were fed restricted diets. In male mice, salicylazosulfapyridine caused an increased incidence of liver neoplasms relative tsms relative to the ad libitum-fed and weight-matched controls. This increase did not occur in the restricted feed protocols. Liver neoplasms in mice are greatly influenced by body weight, and the marked mean body weight reduction observed in dosed male mice in the restricted feed protocols may have overridden the carcinogenic response. Neither t-butylhydroquinone nor scopolamine hydro bromide trihydrate caused increased neoplasm incidences under any of the experimental protocols. Results consistently show that feed restriction caused decreased incidences of neoplasms and nonneoplastic lesions at a variety of anatomic sites in control and dosed animals. Furthermore, the sensitivity of the bioassay to detect a carcinogenic response was altered by dietary restriction: two of the four chemicals caused increased incidences of neoplasms at three sites when evaluated under a standard ad libitum feeding protocol for 104 weeks. When control and dosed groups were subjected to dietary restriction, none of these three sites was detected as a target of carcinogenesis after 2 to 3 years. Rather, one different site of carcinogenesis was detected after 32 months. When dosed animals in the ad libitum feeding protocol were compared to weight-matched control groups, three sites were identified as targets of carcinogenesis and corresponded to the three sites discovered under the ad libitum feeding protocol. The magnitude of the response was greater when the weight-matched controls protocol was used. Dietary restriction of dosed and control animals decreased the sensitivity of these carcinogenesis bioassays. Regarding the future use of dietary restriction regimens in long-term studies, only limited conclusions can be drawn because only four chemicals were evaluated and none of these proved to be a strong carcinogen. However, the results of these studies are consistent with previous findings that dietary restriction increases survival rates and decreases the incidences of neoplasms and nonneoplastic lesions at a variety of sites in rats and mice. This association between reduced body weights and decreased neoplasm incidences underlines the necessity that the doses selected for chronic studies not exceed "minimally toxic doses" so that no marked body weight reductions (or increases) will occur in the dosed groups. Such body weight changes complicate the detection of carcinogenic effects. The following tables summarize and compare the findings from ad libitum-fed, weight-matched, and feed restricted groups for each chemical. Tabular Summary of Dietary Restriction Study of Butyl Benzyl Phthalate is available in web version of this document. TabularSummary of the Dietary Restriction Study of t-Butylhydroquinone is available in web version of this document. TabularSummary of the Dietary Restriction Studies of Salicylazosulfapyridine is available in web version of this document. TabularSummary of the Dietary Restriction Study of Scopolamine Hydrobromide Trihydrate is available in web version of this document. PMID- 12587017 TI - NTP Toxicology and Carcinogenesis Studies of t-Butylhydroquinone (CAS No. 1948-33 0) in F344/N Rats and B6C3F(1) Mice (Feed Studies). AB - t -Butylhydroquinone is used as an antioxidant in cosmetic products such as lipsticks, eye shadows, perfumes, blushers, and skin care preparations at concentrations ranging from 0.1% to 1.0%; the chemical is also used at concentrations up to 0.02% in oils, fats, and meat products to prevent rancidity, and as a polymerization inhibitor for various polyunsaturated polyesters (CIR, 1986). t-Butylhydroquinone was nominated for toxicity and carcinogenicity testing by the Food and Drug Administration. Toxicology and carcinogenicity studies were conducted in F344/N rats and B6C3F(1) mice. Mice were exposed to t-butyl hydroquinone (99% pure) in feed for 13 weeks or 2 years. For rats, exposure to t butylhydroquinone began in utero and continued through lactation. After weaning, pups were fed diets containing the same levels of t-butylhydroquinone as those given to their respective dams for 13 weeks or for up to 30 months. The oral route of administration was selected for these studies because t butylhydroquinone is used as a food additive and human exposure occurs predominantly through this route. In addition to the oral route of exposure, rats were exposed prenatally because perinatal exposure to butylated hydroxytoluene (a structurally related chemical) induced hepatocellular neoplasms in rats. Genetic toxicology studies were conducted in Salmonella typhimurium and cultured Chinese hamster ovary cells in vitro and in mouse bone marrow cells in vivo. 13-WEEK STUDY IN RATS: In the perinatal exposure phase of the 13-week study, groups of 10 female rats (F(0)) were fed 0, 2,500, 5,000, 10,000, 20,000, or 40,000 ppm t butylhydroquinone from 2 weeks prior to cohabitation until the F(1) pups were weaned. F(0) females exposed to 20,000 or 40,000 ppm did not litter. The number of pup deaths in the 5,000 and 10,000 ppm groups was greater than that in the control group, and the average number of surviving pups per litter in the 10,000 ppm group was less than that in the control group. Mean body weights of pups exposed perinatally to 5,000 or 10,000 ppm were lower than that of the controls at the time of weaning. Groups of 10 male and 10 female F(1) rats continued to receive diets containing 0, 2,500, 5,000, or 10,000 ppm t-butylhydroquinone for 13 weeks fol lowing weaning. These dietary levels corresponded to approximately 200, 400, or 800 mg t-butylhydro quinone/kg body weight (males) or 200, 400, or 750 mg/kg (females) per day. All rats survived to the end of the study. The final mean body weights of males and females in the 5,000 and 10,000 ppm groups were significantly lower than those of the controls, as was the mean body weight gain of males exposed to 10,000 ppm. However, interpretation of these findings was complicated by the significantly lower initial mean body weights of the 10,000 ppm groups. Differences in initial body weights were due to in utero exposure to t-butylhydroquinone. Feed consumption by exposed groups of rats was lower than that by controls at week 2, and feed consumption by 5,000 and 10,000 ppm males and 10,000 ppm females was slightly lower than that by controls at the end of the study. Hair discoloration in all exposed groups of rats, except females exposed to 2,500 ppm, was the only clinical observation considered related to chemical exposure. The mean spermatid count, spermatid heads per testis, and spermatid heads per gram of testis were significantly decreased in males exposed to 5,000 ppm. The estrous cycles of females exposed to 2,500 or 5,000 ppm were significantly longer than that of the controls. There were no biologically significant changes in clinical pathology parameters or in organ weights. Increased incidences of hyperplasia of the nasal respiratory epithelium were observed in males exposed to 5,000 ppm and males and females exposed to 10,000 ppm, and an increased incidence of nasal exudate was observed in males in the 10,000 ppm group. Increased incidences of pigmentation were observed in the spleen of male and female rats exposed to 5,000 or 10,000 ppm. Based on lower final mean body weights wer final mean body weights and decreased feed consumption in males and females exposed to 10,000 ppm t-butylhydroquinone, exposure concentrations selected for the long-term rat study were 1,250, 2,500, and 5,000 ppm. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were fed diets containing 0, 2,500, 5,000, 10,000, 20,000, or 40,000 ppm t butylhydroquinone for 13 weeks. These dietary levels corresponded to approximately 440, 880, 1,950, 4,000, and 8,400 mg t-butylhydro quinone/kg body weight (males) or 500, 1,100, 2,200, 4,600, and 9,000 mg/kg body weight (females) per day. There were no exposure-related deaths. Final mean body weights and body weight gains of males and females exposed to 10,000, 20,000, or 40,000 ppm were significantly less than those of the controls. Feed consumption by exposed mice appeared to be similar to that by controls, but there was excessive scatter of feed by mice exposed to 10,000, 20,000, or 40,000 ppm. Therefore, feed consumption by male and female mice in these groups was likely less than that by controls. Significant increases in segmented neutrophil counts occurred at week 3 and at the end of the study in females exposed to 10,000 ppm and males and females exposed to 20,000 or 40,000 ppm. Left caudal, left epididymis, and left testis weights of males exposed to 10,000 or 40,000 ppm were generally significantly lower than those of the controls. The estrous cycle of females exposed to 40,000 ppm was significantly longer than that of the control group. There were no biologically significant differences in organ weights. Increased incidences and severities of mucosal hyperplasia were observed in the forestomach of males exposed to 20,000 or 40,000 ppm and in females exposed to 10,000, 20,000, or 40,000 ppm, and increased incidences of inflammation were observed in the nose and skin of males and females exposed to 10,000, 20,000, or 40,000 ppm. Increased incidences of hyperplasia also occurred in the skin of males and females exposed to 10,000, 20,000, or 40,000 ppm. Based on lower final mean body weights, increased incidences of inflammation of the nose and skin, increased incidences of forestomach mucosal hyperplasia, and increased severity of nonneoplastic lesions observed in mice exposed to 10,000, 20,000, or 40,000 ppm, exposure concentrations selected for the 2-year study were 1,250, 2,500, and 5,000 ppm. LONG-TERM STUDY IN RATS: In the perinatal exposure phase of the long term study, groups of 60 female F(0) rats were fed diets containing 0, 1,250, 2,500, or 5,000 ppm t-butyl hydroquinone, beginning 2 weeks prior to cohabitation and continuing until F(1) pups were weaned. Following weaning, groups of 70 male and 70 female F(1) rats continued to receive diets containing 0, 1,250, or 5,000 ppm, and groups of 68 male and 68 female rats continued to receive diets containing 2,500 ppm. The duration of dosing in feed was 123 weeks post-weaning for males and 129 weeks for females. These exposure concentrations resulted in daily doses of approximately 50, 100, and 200 mg t-butylhydro quinone/kg body weight (males) or 60, 120, and 240 mg/kg (females). Ten male and ten female F(1) rats from each exposure group were evaluated at 3 months. Survival, Body Weights, Feed Consumption, and Clinical Findings: Survival of females exposed to 5,000 ppm was significantly greater than that of the control group. The mean body weights of males and females exposed to 5,000 ppm were generally less than those of the controls throughout the study. Feed consumption by exposed groups was similar to that by controls. Clinical findings of hair discoloration in exposed groups of males and females were considered to be related to chemical exposure. Pathology Findings: No increased neoplasm incidences in male or female rats were attributed to t-butylhydroquinone exposure. The incidences of mammary gland fibroadenoma and fibroadenoma or adenoma (combined) were significantly decreased in males exposed to 1,250 ppm and in all exposed groups of females; and combined incidences of mammary gland fibroadenoma, adenoma, or carcinoma were significantly decreased in all groups of exposed females. The decreases occurred with significant negative trends. Incidences of renal cysts and inflammation were generally increased in exposed groups of male rats. 2-YEAR STUDY IN MICE: Groups of 60 male and 60 female mice received 0, 1,250, 2,500, or 5,000 ppm t-butylhydroquinone in feed for 104 to 105 weeks. These exposure concentrations resulted in daily doses of approximately 150, 300, or 600 mg t-butylhydroquinone/kg body weight (males) or 150, 300, or 700 mg/kg (females). As many as 10 males and 10 females from each exposure group were evaluated at 15 months. Survival, Body Weights and Feed Consumption: Survival of all exposed groups of males and females was similar to that of the control groups. Mean body weights of the 5,000 ppm groups were generally lower than those of the control groups from week 13 until the end of the study. Feed consumption by exposed groups of males and females was similar to that by the controls. There were no biologically significant differences in clinical pathology parameters between control and exposed groups of mice. Pathology Findings: No increased incidences of neoplasms or nonneoplastic lesions in male or female mice were considered to be related to t-butylhydroquinone exposure. GENETIC TOXICOLOGY: t-Butylhydroquinone was not mutagenic in any of four strains of Salmonella typhimurium, with or without liver S9 metabolic activation enzymes. It did, however, induce sister chromatid exchanges and chromosomal aberrations in cultured Chinese hamster ovary cells in the presence, but not the absence, of S9. No increase in the frequency of micronucleated erythrocytes was observed in bone marrow of male mice treated with t butylhydroquinone. CONCLUSIONS: Under the conditions of this long-term feed study, there was no evidence of carcinogenic activity of t-butylhydroquinone in male or female F344/N rats exposed to 1,250, 2,500, or 5,000 ppm. Under the conditions of this 2-year feed study, there was no evidence of carcinogenic activity of t-butylhydroquinone in male or female B6C3F(1) mice exposed to 1,250, 2,500, or 5,000 ppm. Exposure of rats to t-butylhydroquinone in feed resulted in decreased incidences of mammary gland neoplasms in males and females. Synonyms: Tert-butyl-hydroquinone; 2-(1,1-dimethylethyl)-1,4-benzenediol; 2-tert butylhydroquinone; mono-tert-butylhydroquinone; tert-butyl-1,4-benzenediol: mono tertiarybutylhydroquinone; 2-tert-butyl-1,4-benzenediol; 2-(1,1 dimethyl)hydroquinone; 2-(tert-butyl)-p-hydroquinone; TBHQ; MTBHQ Trade Names: Sustane; Tenox TBHQ; Banox 20BA PMID- 12587018 TI - NTP Toxicology and Carcinogenesis Studies of Butyl Benzyl Phthalate (CAS No. 85 68-7) in F344/N Rats (Feed Studies). AB - Butyl benzyl phthalate is a plasticizer added to polymers to give flexibility and softness. It is used extensively in polyvinyl chloride and in cellulose plastics, polyvinyl acetate, polysulfides, and polyurethane. Butyl benzyl phthalate was nominated as part of a class study of phthalates. Previous studies of butyl benzyl phthalate by the NTP (1982a) resulted in chemical-related mortality in male rats beginning at about 14 weeks of exposure and, thus, were inadequate for evaluating carcinogenicity in male rats. The companion studies revealed a marginal increase in leukemia in female rats and no evidence of carcinogenicity in B6C3F1 mice. Consequently, the present evaluations were conducted only in F344/N rats. Male and female F344/N rats were given butyl benzyl phthalate (at least 97% pure) in feed for 10 weeks, 26 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, L5178Y mouse lymphoma cells, cultured Chinese hamster ovary cells, mouse bone marrow cells, and Drosophila melanogaster. 10-WEEK MODIFIED MATING STUDY IN RATS: Groups of 15 male F344/N rats were given 0, 300, 2,800, or 25,000 ppm butyl benzyl phthalate (equivalent to average daily doses of approximately 20, 200, or 2,200 mg butyl benzyl phthalate/kg body weight) in feed for 10 weeks. All rats survived to the end of the study. The final mean body weight and body weight gain of the 25,000 ppm group were significantly less than those of the controls. Feed consumption by the 25,000 ppm group was less than that by the controls at the end of the study. A few minimal hematology changes occurred in the 25,000 ppm male rats. There was some evidence of a minimal anemia characterized by a decreased erythrocyte count and increases in mean cell hemoglobin and platelet count. The absolute and relative prostate gland weights of the 25,000 ppm males were significantly less than those of the controls. Degeneration of the seminiferous tubule germinal epithelium was observed in all males from the 25,000 ppm group. The absolute right cauda, right epididymis, and right testis weights of the 25,000 ppm males were significantly less than those of the controls. The epididymal spermatozoal concentrations in 2,800 and 25,000 ppm males were significantly less than that in the controls. Although 10 females mated to 25,000 ppm males were initially found to be sperm positive, none of these females were pregnant at necropsy. The fertility indices of males and females in the 25,000 ppm group were significantly lower than those of the controls. The maternal body weights of females mated to 300 and 2,800 ppm males were similar to those of females mated to control males. There were no significant differences in litter data between the controls and the 300 and 2,800 ppm groups. 26-WEEK STUDY IN RATS: Groups of 15 male F344/N rats were given 0, 300, 900, 2,800, 8,300, or 25,000 ppm butyl benzyl phthalate in feed for 26 weeks. Dietary levels of 300, 900, 2,800, and 8,300 ppm delivered average daily doses of approximately 30, 60, 180, and 550 mg butyl benzyl phthalate/kg body weight. The final mean body weight and body weight gain of the 25,000 ppm males were significantly less than those of the controls. Except for the 25,000 ppm males, feed consumption by all exposed groups was similar to that by the controls. An exposure-related macrocytic responsive anemia was present in the 25,000 ppm group at all time points. Additionally, minimal erythrocyte count decreases occurred sporadically in the 2,800 and 8,300 ppm groups at various time points. Reticulocyte counts were increased on days 60 and 90. Increases in mean cell hemoglobin and mean cell hemoglobin concentrations occurred in the 8,300 and 25,000 ppm rats. The absolute right cauda, right epididymis, and right testis weights and the sperm concentration of 25,000 ppm males were significantly less than those of the controls. The incidences of hypospermia and of atrophy of the seminiferous tubule in the testis and of hypospermia in the epididymis in 25,000 ppm males were significantly greater than those in the in the controls. Degenerative changes of the testis and epididymis in the 25,000 ppm males were qualitatively and quantitatively similar to those observed in males in the 10 week modified mating study. 2-YEAR STUDY IN RATS: Groups of 60 male F344/N rats were given 0, 3,000, 6,000, or 12,000 ppm butyl benzyl phthalate (equivalent to average daily doses of approximately 120, 240, or 500 mg butyl benzyl phthalate/kg body weight), and groups of 60 female F344/N rats were given 0, 6,000, 12,000, or 24,000 ppm butyl benzyl phthalate (equivalent to average daily doses of approximately 300, 600, or 1,200 mg/kg) in feed for 2 years. Survival, Body Weights, and Feed Consumption: Survival of all exposed groups of male and female rats was similar to that of the controls. Mean body weights of the 12,000 ppm males and 24,000 ppm females were less than those of the controls throughout most of the study. Feed consumption by the females exposed to 24,000 ppm was less than that by the controls at the beginning of the study, but was similar to that by the controls by week 6. Hematology and Hormone Assays: In general, hematology changes were sporadic and minor. At 6 months, a minimal decrease in erythrocyte count and an increase in mean cell hemoglobin, similar to that which occurred in the 26-week study, occurred in male rats in the 12,000 ppm group. In female rats, a decreased hematocrit value occurred at 15 months in the 24,000 ppm group. There was also a mild decrease in triiodothyronine concentrations in the 24,000 ppm females at 6 and 15 months and at the end of the study. Pathology Findings: of pancreatic acinar cell adenoma and adenoma or carcinoma (combined) in 12,000 ppm males were significantly greater than those in the controls. The incidences of adenoma and of adenoma or carcinoma (combined) in 12,000 ppm males exceeded the ranges of historical controls from NTP 2-year feed studies. One carcinoma was observed in one 12,000 ppm male, and two adenomas were observed in 24,000 ppm females. At 2 years, the incidence of focal hyperplasia of the pancreatic acinar cell in 12,000 ppm males was significantly greater than that in the controls. At 2 years, transitional epithelial papillomas in the urinary bladder were observed in one control female and in two 24,000 ppm females. The incidence of this neoplasm exceeded the range of historical controls from NTP 2-year feed studies. The incidence of transitional epithelial hyperplasia in 24,000 ppm females was significantly greater than that in the controls. The absolute right kidney weight of 12,000 ppm females and the relative right kidney weights of all exposed groups of males and of 24,000 ppm females were significantly greater than those of the controls at the 15-month interim evaluation. The severities of renal tubule pigmentation in 12,000 ppm males and in 24,000 ppm females were greater than those in the controls at 15 months and 2 years. At 2 years, the incidences of kidney mineralization in 6,000 and 24,000 ppm females were significantly less than that in the controls, and the severity was decreased in exposed females. The incidence of preputial gland adenoma or carcinoma (combined) in 12,000 ppm male rats was significantly less than in the controls, and the incidences occurred with a negative trend. GENETIC TOXICOLOGY: Results from in vitro mutagenicity tests with butyl benzyl phthalate were uniformly negative. No mutagenic response was obtained in any of several strains of Salmonella typhimurium treated with up to 11,550 mg/plate butyl benzyl phthalate, with or without S9 metabolic activation enzymes. Negative results were also obtained in in vitro studies of mammalian cell systems with and without S9. No induction of trifluorothymidine resistance in L5178Y mouse lymphoma cells or sister chromatid exchanges and chromosomal aberrations in cultured Chinese hamster ovary cells were observed. These assays also were conducted with and without S9. No significant increase in sex-linked recessive lethal mutations was observed in germ cells of male Drosophila melanogaster after administration of butyl benzyl phthalate either in feed or by injection. In contrast to the negative results obtained in vitro and in Drosophila, butyl benzyl phthalate gave positive responses in two in vivo studies with mice. Results of a mouse bone marrow sister chromatid exchange test were positive at sample times of 23 and 42 hours, but no confirmatory test was conducted. Chromosomal aberrations were induced in bone marrow cells of male mice sampled 17 hours after intraperitoneal injection of 5,000 mg/kg butyl benzyl phthalate. CONCLUSIONS: Under the conditions of this 2-year feed study, there was some evidence of carcinogenic activity of butyl benzyl phthalate in male F344/N rats based on the increased incidences of pancreatic acinar cell adenoma and of acinar cell adenoma or carcinoma (combined). There was equivocal evidence of carcinogenic activity of butyl benzyl phthalate in female 344/N rats based on the marginally increased incidences of pancreatic acinar cell adenoma and of transitional epithelial papilloma of the urinary bladder. Exposure of rats to butyl benzyl phthalate in feed for 2 years resulted in focal hyperplasia in the pancreas in male rats and in transitional epithelial hyperplasia in the urinary bladder of female rats. Synonyms: A13-14777; BBP; 1,2-benzenedicarboxylic acid butyl phenylmethyl ester (9CI); benzyl n-butyl phthalate; n-butyl benzyl phthalate; butyl phenylmethyl 1,2-benzenedicarboxylate; NCI-C54375; phthalic acid benzyl butyl ester (8CI) Trade names: Palatinol BB; Santicizer 160; Sicol 160; Unimoll BB PMID- 12587019 TI - NTP Toxicology and Carcinogenesis Studies of Salicylazosulfapyridine (CAS No. 599 79-1) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Salicylazosulfapyridine is widely used for the treatment of ulcerative colitis and Crohn's disease. It has been beneficial in the treatment of psoriasis and rheumatoid arthritis, and it has been used in veterinary medicine for the treatment of granulomatous colitis. Salicylazosulfapyridine was nominated for toxicity and carcinogenicity testing by the National Cancer Institute on the basis of its widespread use in humans and because it is a representative chemical from a class of aryl sulfonamides. Salicylazosulfapyridine is a suspect carcinogen because reductive cleavage of the azo linkage yields a p-amino aryl sulfonamide (sulfapyridine), and a related p-amino aryl sulfonamide (sulfamethoxazole) has been shown to produce thyroid neoplasms in rats. Toxicology and carcinogenicity studies were conducted in F344/N rats and B6C3F1 mice. Rats and mice were administered salicylazosulfapyridine (96% to 98% pure) in corn oil by gavage for 16 days, 13 weeks, or 2 years. The gavage route of administration was selected for these studies because it approximates the typical route of human exposure to the chemical. Genetic toxicology studies were conducted in vitro in Salmonella typhimurium and cultured Chinese hamster ovary cells and in vivo in rat and mouse bone marrow and mouse peripheral blood cells. 16-DAY STUDY IN RATS: Groups of five male and five female rats were administered 0, 675, 1,350, or 2,700 mg salicylazosulfapyridine/kg body weight in corn oil by gavage for 16 days excluding weekends. All rats survived to the end of the study. With the exception of the 675 mg/kg male group, the final mean body weights of all dosed groups of males and females were significantly lower than those of controls. Mean body weight gains of all dosed groups were less than those of controls. Clinical findings included ruffled fur and distended abdomens in male and female rats receiving 2,700 mg/kg. Hypothyroidism, evidenced by decreased serum triiodothyronine and thyroxine concentrations and increased thyroid stimulating hormone concentrations, occurred in 2,700 mg/kg male and female rats. The absolute and relative thymus weights of male rats receiving,350 or 2,700 mg/kg and female rats receiving 2,700 mg/kg were significantly lower than those of controls. At necropsy, all dosed rats had enlarged cecae/large intestines. Male rats receiving 1,350 mg/kg and male and female rats receiving 2,700 mg/kg had red, enlarged thyroid glands. Chemical-related microscopic lesions were present in the forestomach, thymus, thyroid gland, and pituitary gland. Minimal to mild hyperplasia of the forestomach mucosa was present in the 1,350 and 2,700 mg/kg male and female groups. Lymphoid depletion was observed in the thymus of three male and three female rats in the 2,700 mg/kg groups. Male and female rats receiving 1,350 and 2,700 mg/kg had thyroid gland follicular cell hyperplasia and an increase in thyroid-stimulating hormone producing cells in the pars distalis of the pituitary gland. 16-DAY STUDY IN MICE: Groups of five male and five female mice were administered 0, 675, 1,350, or 2,700 mg salicylazosulfapyridine/kg body weight in corn oil by gavage for 16 days excluding weekends. There were no chemical-related deaths, and final mean body weights of dosed mice were similar to those of controls. No chemical-related clinical findings were noted for male or female mice. There were no differences in triiodothyronine, thyroxine, or thyroid-stimulating hormone concentrations between dosed and control mice. There were no biologically significant differences in absolute or relative organ weights between dosed and control male and female mice. At necropsy, male mice receiving 2,700 mg/kg had enlarged cecae/large intestines. There were no biologically significant histopathologic lesions attributed to salicylazosulfapyridine administration. 13-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were administered 0, 84, 168.8, or 337.5 mg salicylazosulfapyridine/kg body weight in corn oil by gavage for 13 weeks. All rats survived to the end of the study. The finaludy. The final mean body weights of dosed male rats were similar to those of controls; the final mean body weights and body weight gains of dosed females were significantly lower than those of controls. No chemical-related clinical findings were noted in dosed male or female rats during the 13-week study. No significant differences in hematology or urinalysis parameters between control and dosed rats were observed. The absolute and relative right kidney weights of 337.5 mg/kg females were significantly greater than those of controls. At necropsy, some 337.5 mg/kg male rats had red, enlarged thyroid glands. Histopathologic changes were noted primarily in the thyroid gland and pituitary gland of males and females in the 337.5 mg/kg groups. The thyroid gland lesions observed were similar to those present in the 16-day study. Nine male rats receiving 168.8 mg/kg and ten male and seven female rats receiving.5 mg/kg had minimal but consistent changes in thyroid gland follicular cells. In the pituitary gland of 337.5 mg/kg males and females, the thyroid stimulating hormone producing cells were enlarged and contained pale-staining cytoplasm and prominent Golgi complexes. Decreased serum triiodothyronine and thyroxine concentrations and increased thyroid-stimulating hormone concentration, similar to differences observed in the 16-day study, occurred in 337.5 mg/kg male rats; thyroid hormone concentrations were not affected in female rats. Sperm motility of all dosed groups of males was significantly lower than that of controls. Vaginal cytology parameters of dosed groups of females were similar to those of controls. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were administered 0, 675, 1,350, or 2,700 mg salicylazosulfapyridine/kg body weight in corn oil by gavage for 13 weeks. All mice survived to the end of the study. The final mean body weights of dosed male and female mice were similar to those of controls. The mean body weight gains of 1,350 and 2,700 mg/kg male mice were less than that of controls. No chemical-related clinical findings were noted in dosed male or female mice during the 13-week study. There was minimal evidence of a responsive anemia in mice in the 13-week study. The anemia was probably related to a methemoglobinemia. There were minimal decreases in thyroxine concentration in all dosed groups of male and female mice in the -week study. There were, however, no differences in triiodothyronine and thyroid-stimulating hormone concentrations between dosed and control animals. Absolute and relative liver weights of all groups of dosed male and female mice were significantly greater than those of controls. There were no chemical-related gross lesions. Microscopic evaluation of the liver revealed centrilobular hypertrophy in five 1,350 mg/kg and all 2,700 mg/kg male mice. The right cauda weight of the 1,350 mg/kg group and the right epididymis weights of all dose groups were significantly lower than those of controls. There was no evidence of chemical related alteration in the vaginal cytology parameters of female mice. 2-YEAR STUDY IN RATS: Groups of 60 male and 60 female rats were administered 84, 168, or 337.5 mg salicylazosulfapyridine/kg body weight in corn oil by gavage for up to 105 weeks. Groups of 70 male and 60 female rats were administered the corn oil vehicle by gavage for up to 105 weeks. A stop-exposure group of 70 male rats was administered 337.5 mg/kg salicylazosulfapyridine in corn oil by gavage for 6 months, after which animals received the corn oil vehicle by gavage for the remainder of the 2-year study. Ten animals from the vehicle control male group and 10 animals from the 337.5 mg/kg stop-exposure group were evaluated at 6 months; animals from each core-study group were evaluated at 15 months. Survival, Body Weights, and Clinical Chemistry: Survival of 337.5 mg/kg male core-study rats was significantly lower than that of controls; survival of 84 and 168 mg/kg core-study males, all groups of dosed females, and the stop-exposure male group was similar to controls. Mean body weights of core-study males and stop-exposure males were similar to controls throughout the study. From week 45 to the end of the study, females in the 337.5 mg/kg group had mean body weights that were lower than those of controls. The serum thyroxine concentration in 337.5 mg/kg core study males at study termination was minimally lower than that of controls; the serum thyroid-stimulating hormone, triiodothyronine, and reverse triiodothyronine concentrations of dosed males and females were similar to those of controls. Pathology Findings: Administration of salicylazosulfapyridine for 2 years was associated with transitional epithelial papilloma in the urinary bladder of male rats and may have been associated with transitional epithelial papilloma of the kidney and of the urinary bladder of female rats. Nonneoplastic effects in the urinary bladder and kidney of male and female rats and in the spleen of male rats were also observed. Dosed male and female rats had increased incidences of grossly and microscopically observed urinary bladder concretions (diagnosed grossly as calculi at necropsy); male and female rats that developed transitional epithelial papillomas of the urinary bladder had grossly observed concretions (calculi) in the urinary bladder at necropsy. The microscopic neoplastic and nonneoplastic urinary bladder and kidney effects observed in dosed male rats during the 2-year continuous study did not occur in dosed rats during the 2-year stop-exposure study, nor were there gross observations of concretions (calculi) at necropsy. The incidences of mononuclear cell leukemia in male and female rats were decreased. The thyroid gland hyperplasia seen in the -week study was not observed in the 2-year study, and there was no evidence of chemical-related thyroid gland follicular cell adenomas or carcinomas. 2-YEAR STUDY IN MICE: Groups of 60 male and 60 female mice were administered 0, 675, 1,350, or 2,700 mg salicylazosulfapyridine/kg body weight in corn oil by gavage for up to 104 weeks. Ten animals from each group were evaluated at 15 months. Survival, Body Weights,and Clinical Chemistry: Survival of all the dosed groups of male and female mice was similar to that of controls. Mean body weights of 675 and 1,350 mg/kg male and female mice were similar to controls throughout the study. From week 12 to the end of the study, 2,700 mg/kg male mice had mean body weights that were lower than those of controls. From week 14 to the end of the study, the 2,700 mg/kg female mice had mean body weights that were lower than those of controls. There were no chemical-related differences in triiodothyronine, reverse triiodothyronine, thyroxine, or thyroid-stimulating hormone concentrations between dosed and control mice at the 15-month evaluation. Pathology Findings: Exposure of mice to salicylazosulfapyridine in corn oil by gavage for 2 years was associated with increased incidences of hepatocellular neoplasms in males and females. Nonneoplastic effects in the liver and spleen were also observed in male and female mice. The incidences of forestomach squamous cell papilloma in females and forestomach hyperplasia in males and females were decreased. GENETIC TOXICOLOGY: Salicylazosulfapyridine was not mutagenic in Salmonella typhimurium strains TA97, TA98, TA100, or TA1535, and it did not induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells. These in vitro assays were performed with and without S9 metabolic activation enzymes. Results from in vivo mouse bone marrow chromo somal aberration tests were uniformly negative, while results of micronucleus assays performed on male or female mice exposed to salicylazosulfapyridine for periods ranging from 3 days to weeks were positive. Micronucleus tests in male mice for shorter exposure times (1 to 2 days) yielded negative or very weakly positive results. A three treatment (72-hour exposure time) micronucleus test performed in male rats yielded equivocal results. Overall, results of these in vivo assays indicate that salicylazosulfa pyridine is capable of inducing chromosomal damage, possibly in the form of aneuploidy, in mouse bone marrow cells after multiple administrations. CONCLUSIONS: Under the conditions of these 2-year gavage studies, there was some evidence of carcinogenic activity of salicylazosulfapyridine in male and female F344/N rats based on increased incidences of neoplasms in the urinary tract. There was an increased incidence of transitional epithelial papilloma of the urinary bladder in males and a low incidence of rare transitional epithelial papillomas of the kidney and of the urinary bladder in females. There was clear evidence of carcinogenic activity of salicylazosulfapyridine in male and female B6C3F1 mice based on increased incidences of hepatocellular neoplasms. Increased incidences of nonneoplastic lesions of the urinary bladder and kidney in male and female rats and of the spleen in male rats were observed. Increased incidences of nonneoplastic lesions of the liver and spleen in male and female mice were observed. Decreased incidences of mononuclear cell leukemia in male and female rats were related to salicylazosulfapyridine administration. Decreased incidences of forestomach squamous cell papilloma in female mice and forestomach hyperplasia in male and female mice were related to salicylazosulfapyridine administration. Synonyms: 2 Hydroxy-5-[[4-[2-(pyridinylamino)sulfonyl]phenyl]azo]benzoic acid; 5-[p- (2 pyridylsulfamoyl)phenylazo]salicylic acid; sulfasalazine; salazosulfapyridine; 5 [4-(2-pyridylsulfamoyl)phenylazo]-2-hydroxybenzoic acid; 4-(pyridyl-2 amidosulfonyl)-3'-carboxy-4'-hydroxyazobenzene; sulphasalazine Trade names: Azopyrin, Azulfidine, Benzosulfa, Colo-Pleon, Reupirin, Salazopyrin PMID- 12587020 TI - NTP Toxicology and Carcinogenesis Studies of 1,2-Dihydro-2,2,4-Trimethylquinoline (CAS No. 147-47-7) in F344/N Rats and B6C3F1 Mice (Dermal Studies) and the Dermal Initiation/Promotion Study in Female Sencar Mice. AB - 1,2-Dihydro-2,2,4-trimethylquinoline (monomer) is used as an antioxidant in styrene-butadiene and nitrile-butadiene rubbers and latexes. It was nominated by the National Cancer Institute as part of a review of chemicals used in the manufacture and processing of rubber, during which potential occupational and consumer exposure to this compound can occur. It was selected for evaluation because it is a derivative of quinoline, a known rodent carcinogen, and was regarded as having potential carcinogenic activity. Because of the pattern of use and exposure, dermal administration was considered most appropriate. Male and female F344/N rats and B6C3F1 mice received topical applications of 1,2-dihydro 2,2,4-trimethylquinoline in acetone (greater than 90% pure) for 13 weeks or 2 years. Groups of female SENCAR mice received 1,2-dihydro-2,2,4-trimethylquinoline (greater than 90% pure) during a 1-year dermal initiation/promotion study to determine the tumor initiation or promotion potential of the chemical. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, and mouse peripheral blood cells. 13-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were topically administered 0, 5, 20, 50, 100, or 200 mg 1,2-dihydro-2,2,4-trimethylquinoline/kg body weight in acetone, 5 days per week for 13 weeks. In addition, there were 10 male and 10 female untreated controls. All rats survived to the end of the study. Final mean body weights and mean body weight gains of treated male and female rats were similar to those of the vehicle controls except those of 200 mg/kg males, which were significantly lower than those of the vehicle controls. The only notable clinical observation was skin discoloration of treated rats. In the 200 mg/kg groups, absolute and relative liver weights of males and absolute liver weights of females were significantly greater than those of the vehicle controls. There were no significant differences in hematology or clinical chemistry parameters, reproductive tissue parameters, or estrous cycle characterization between treated and control groups. Histopathologic lesions of the skin at the site of application included acanthosis and hyperkeratosis in 100 and 200 mg/kg males and 200 mg/kg females. Cytoplasmic vacuolization of hepatocytes of mild to moderate severity was observed in the livers of all 200 mg/kg males and was considered treatment related. Based on the incidence and severity of skin and liver lesions observed in 200 mg/kg rats in the 13-week study, 100 mg/kg was selected as the high dose for the 2-year rat study. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were topically administered 0, 2.5, 5, 10, 20, or 50 mg 1,2 dihydro-2,2,4-trimethylquinoline/kg body weight in acetone, 5 days per week for 13 weeks. In addition, there were 10 male and 10 female untreated controls. All mice except one 2.5 mg/kg female survived to the end of the study. Final mean body weights and mean body weight gains of male and female mice were similar to those of the vehicle controls. There were no treatment-related clinical observations. There were no significant differences between treated and control groups in organ weights, hematology and clinical chemistry parameters, reproductive tissue parameters, or estrous cycle characterization. Histopathologic lesions of the skin at the site of application included acanthosis (epidermal hyperplasia), hyperkeratosis, and parakeratosis, all ranging from minimal to mild in severity. Minimal to mild fibrosis and subchronic inflammation were observed in the dermis. Based on the incidences and severities of skin lesions observed in 20 and 50 mg/kg mice in the 13-week study, 10 mg/kg was selected as the high dose for the 2-year mouse study. 2-YEAR STUDY IN RATS: Groups of 60 male and 60 female F344/N rats were topically administered 0, 36, 60, or 100 mg 1,2-dihydro-2,2,4-trimethylquinoline/kg body weight in acetone, 5 days per week for 103 (males) or 104 (females) weeks. Ten rats per group were evaluated after 15 moed after 15 months of treatment. Survival and Body Weights Survival of treated rats was similar to that of controls. Mean body weights of 60 mg/kg males and 100 mg/kg males and females were slightly lower than those of the controls after week 21. Mean body weights of 36 mg/kg males and females and 60 mg/kg females were generally similar to those of the controls throughout the study. Pathology Findings: No skin neoplasms were attributed to treatment with 1,2-dihydro-2,2,4-trimethylquinoline. Several nonneoplastic skin lesions were determined to be treatment related. Incidences of acanthosis at the site of application in all treated groups of males and in 100 mg/kg females at the 15 month interim evaluation were significantly greater than those in the controls. At the end of the 2-year study, incidences of acanthosis at the site of application in 60 and 100 mg/kg males and females and hyperkeratosis at the site of application in 60 mg/kg females were significantly greater than those in the controls. Absolute and relative right kidney weights of 60 and 100 mg/kg male rats were significantly greater than those of the controls at the 15-month interim evaluation. Incidences of renal tubule adenoma and adenoma or carcinoma (combined) in all treated groups of males were significantly greater than those in the controls. These incidences exceeded the range from the historical controls in 2-year NTP feed studies. An extended (step section) evaluation of the kidneys of male rats did not reveal an additional increase in neoplastic response because additional adenomas and hyperplasias were observed in the controls as well as in treated groups. 2-YEAR STUDY IN MICE: Groups of 60 male and 60 female B6C3F1 mice were topically administered 0, 3.6, 6, or 10 mg 1,2-dihydro-2,2,4 trimethylquinoline/kg body weight in acetone, 5 days per week for 103 (males) or 104 (females) weeks. Nine or ten mice per group were evaluated after 15 months of treatment. Survival and Body Weights: Survival of treated mice was similar to that of controls. Mean body weights of treated male and female mice were similar to those of the controls throughout the study. Pathology Findings: No neoplasms or nonneoplastic lesions were attributed to treatment with 1,2-dihydro-2,2,4 trimethylquinoline. 1-YEAR INITIATION/PROMOTION STUDY IN FEMALE SENCAR MICE: Groups of 30 female SENCAR mice were topically administered varying initiation/promotion treatments as outlined in the table below. Survival, Body Weights, and Clinical Findings: Survival in all treated groups was similar to that of the respective controls, except in the 2.5 mg 7,12 dimethylbenz(a)anthracene (DMBA)/0.5 mg 12-O-tetradecanoylphorbol-13-acetate (TPA) group in which survival was significantly lower than that of the controls. Mean body weights of all treated groups were similar to those of the respective controls throughout the study. No clinical observations were associated with 1,2 dihydro-2,2,4-trimethylquinoline treatment; however, mice promoted with TPA showed signs of irritation and papilloma at the site of application. Pathology Findings: Initiation and promotion with acetone alone was not associated with any skin lesions at the site of application. The incidences of acanthosis and chronic inflammation were increased in all groups promoted with TPA regardless of the initiator treatment; however, the incidences of nonneoplastic lesions were low in all other groups. Incidences of squamous cell papillomas and squamous cell carcinomas were markedly increased in the DMBA/TPA positive control group; however, no response was observed in groups initiated with DMBA and promoted with 5, 10, or 25 mg/kg 1,2-dihydro-2,2,4-trimethylquinoline or in the group initiated with 1,2-dihydro-2,2,4-trimethylquinoline and promoted with TPA. GENETIC TOXICOLOGY: 1,2-Dihydro-2,2,4-trimethylquinoline was not mutagenic in any of several strains of Salmonella typhimurium, with or without S9 metabolic activation. 1,2-Dihydro-2,2,4-trimethylquinoline induced sister chromatid exchanges in cultured Chinese hamster ovary cells in the absence of S9, but not in the presence of S9. However, no increase in the frequency of chromosomal aberrations was observed in cultured Chinese hamster ovary cells treated with 1,2 dihydro-2,2,4-trimethylquinoline, with or without S9. No increase in the frequency of micronucleated erythrocytes was noted in peripheral blood of male or female mice exposed topically to 1,2-dihydro-2,2,4-trimethylquinoline for 13 weeks. CONCLUSIONS: Under the conditions of these 2-year dermal studies, there was some evidence of carcinogenic activity of 1,2-dihydro-2,2,4 trimethylquinoline in male F344/N rats, based on increased incidences of renal tubule adenoma and adenoma or carcinoma (combined). There was no evidence of carcinogenic activity of 1,2-dihydro-2,2,4-trimethylquinoline in female F344/N rats receiving 36, 60, or 100 mg/kg, or in male or female B6C3F1 mice receiving 3.6, 6, or 10 mg/kg. Exposure of rats to 1,2-dihydro-2,2,4-trimethylquinoline by dermal application in acetone for 2 years resulted in acanthosis in males and females and hyperkeratosis in females at the site of application. No nonneoplastic lesions in male or female mice were attributed to treatment with 1,2-dihydro-2,2,4-trimethylquinoline. Synonyms: 2,2,4-Trimethyl-1,2 dihydroquinoline; acetone anil; methylquinoline Trade names: Agerite Resin D; Flectol A; Flectol H; Flectol Pastilles; Vulkanox HS/LG; Vulkanox HS/Powder PMID- 12587021 TI - NTP Toxicology and Carcinogenesis Studies of Codeine (CAS No. 76-57-3) in F344 Rats and B6C3F1 Mice (Feed Studies). AB - Codeine is used in a variety of pharmaceuticals including analgesics, sedatives, hypnotics, antiperistaltics, and antitussive agents. The National Cancer Institute and the Food and Drug Administration nominated codeine for study because it is a widely used drug and it is representative of the morphine class of compounds, for which chronic carcinogenicity studies had not been conducted. The oral route of administration was selected because it is the primary route of human exposure. Male and female F344/N rats and B6C3F1 mice were given codeine (99% pure) in feed for 14 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and cultured Chinese hamster ovary cells. 14-DAY STUDY IN RATS: Groups of five male and five female F344/N rats were given 0, 1,562, 3,125, 6,250, 12,500, or 25,000 ppm codeine in feed for 14 days, which resulted in daily doses of approximately 125, 250, 450, 650, or 750 mg codeine/kg body weight to males and 125, 250, 500, 700, or 300 mg/kg to females. One female exposed to 6,250 ppm, one male and three females exposed to 12,500 ppm, and all males and females exposed to 25,000 ppm died during the study. Final mean body weights and mean body weight gains of all exposed groups except 1,562 ppm females were significantly lower than those of the controls. No chemical related gross lesions were observed in rats at necropsy. Thickening of the forestomach mucosa (hyperplasia and hyperkeratosis) and lymphoid depletion of the thymus in exposed males and females and testicular degeneration in exposed males, observed primarily in the 12,500 and 25,000 ppm groups, were associated with decreased survival and increased morbidity in these groups. 14-DAY STUDY IN MICE: Groups of five male and five female B6C3F1 mice were given 0, 781, 1,562, 3,125, 6,250, or 12,500 ppm codeine in feed for 14 days, which resulted in daily doses of approximately 150, 300, 600, 1,300, or 3,000 mg codeine/kg body weight to males and 200, 400, 750, 1,500, or 3,000 mg/kg to females. All mice survived to the end of the study. The final mean body weight of 3,125 ppm females was significantly greater than that of the controls; the final mean body weight of 12,500 ppm females and the mean body weight gains of 12,500 ppm males and females were significantly lower than those of the controls. Absolute and relative liver weights of 3,125, 6,250, and 12,500 ppm males and of 12,500 ppm females and the absolute and relative right kidney weights of 12,500 ppm males were significantly lower than those of the controls. No gross or histopathologic lesions were attributed to codeine exposure. 13-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were given 0, 390, 781, 1,562, 3,125, or 6,250 ppm codeine in feed for 13 weeks, which resulted in daily doses of approximately 25, 50, 100, 200, or 450 mg codeine/kg body weight to males and 25, 50, 100, 250, or 500 mg/kg to females. There were no chemical-related deaths during the study. Final mean body weights and mean body weight gains of all groups of exposed males and of females exposed to 1,562, 3,125, and 6,250 ppm were significantly lower than those of the controls. Feed consumption decreased with increasing exposure concentration during the first week of the study; however, by the end of the study, feed consumption by most exposed groups was similar to that by the controls. There were alterations of various hematology and clinical chemistry parameters at the end of the study. There was a mild dose-dependent lymphopenia in females receiving 1,562 ppm and above and in 6,250 ppm males. There also was a minimal to mild macrocytosis that occurred in all exposed groups of males and in females exposed to 781, 3,125, or 6,250 ppm. No significant differences between control and exposed rats were observed in sperm morphology or vaginal cytology parameters. Absolute and relative adrenal gland weights of exposed males and of 3,125 and 6,250 ppm females were significantly greater than those of the controls. Absolute and relative liver weights of exposed males werees were significantly lower than those of the controls. Relative thymus weights of 3,125 and 6,250 ppm males were significantly lower than that of the controls. No chemical-related gross or histopathologic lesions were observed in male or female rats. 13-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were given 0, 390, 781, 1,562, 3,125, or 6,250 ppm codeine in feed for 13 weeks, which resulted in daily doses of approximately 60, 120, 260, 460, or 1,000 mg codeine/kg body weight to males and 60, 130, 280, 530, or 1,200 mg/kg to females. Two male mice in the 3,125 ppm group died during week 7. All other mice survived to the end of the study. Final mean body weights of exposed males and females were similar to those of the controls. Feed consumption by exposed males and females was similar to that by the controls. Abnormal posture was observed in all exposed groups of males. There were no significant differences in hematology or urinalysis parameters in male or female mice. Minor, sporadic changes occurred in a few of the clinical chemistry parameters; they were not considered biologically significant. No significant differences in sperm morphology or vaginal cytology were attributed to codeine exposure. Absolute and relative kidney weights of 3,125 and 6,250 ppm males were lower than those of the controls. No chemical related differences in organ weights were observed in females. No chemical related gross or histopathologic lesions were observed in male or female mice. 2 YEAR STUDY IN RATS: Groups of 60 male and 60 female F344/N rats were fed diets containing 0, 400, 800, or 1,600 ppm codeine for up to 106 weeks, with 9 or 10 rats per group evaluated at 15 months. These exposure concentrations resulted in average daily doses of approximately 15, 30, and 70 mg codeine/kg body weight to males and 15, 40, and 80 mg/kg to females. Survival, Body Weights, Feed Consumption, and Clinical Findings Survival of 400 ppm females was significantly greater than that of the controls; survival of all groups of exposed males and of 800 and 1,600 ppm females was similar to that of the controls. There was an exposure-related decrease in mean body weights of males and females. The final mean body weight of 1,600 ppm males was 88% that of the controls, and the final mean body weight of 1,600 ppm females was 89% that of the controls. Feed consumption by exposed groups was similar to that by the controls. Chemical related clinical findings were limited to ocular discharge in exposed males and females. Pathology Findings: Absolute and relative adrenal gland weights of 800 and 1,600 ppm males were significantly greater than those of the controls at 15 months. There were no increased incidences of neoplasms attributable to codeine exposure at any site. At 2 years, there were exposure-related decreases in the incidences of adrenal medulla hyperplasia in males and females. There was an exposure-related decrease in the incidence of benign pheochromocytomas in males, and the incidences in exposed males were significantly lower than that in the controls. In 1,600 ppm females the incidences of mammary gland fibroadenomas and of fibroadenomas or adenocarcinomas (combined) were significantly lower than those in the controls. The decreased incidences of benign pheochromocytomas in males and mammary gland neoplasms in females were considered to be related to codeine exposure. 2-YEAR STUDY IN MICE: Groups of 60 male and 60 female B6C3F1 mice were fed diets containing 0, 750, 1,500, or 3,000 ppm codeine for up to 106 weeks, with 9 or 10 mice per group evaluated at 15 months. These exposure concentrations resulted in average daily doses of approximately 100, 200, or 400 mg codeine/kg body weight to males and females. Survival, Body Weights, Feed Consumption, and Clinical Findings: Survival of exposed males and females was similar to that of the controls. Mean body weights of 750 and 1,500 ppm males and females were similar to those of the controls throughout most of the study. Mean body weights of 3,000 ppm males and females were less than those of the controls from about week 13, and the final mean body weights of these groups were 86% and 82% those of the respective controls. Feed consumption by exposed groups was similar to that by the controls. Pathology Findings: There were no increased incidences of neoplasms attributable to codeine exposure at any site. At 15 months, the incidence of thyroid gland follicular cell hyperplasia in 3,000 ppm males was significantly greater than that of the controls, and this lesion was observed in 1,500 and 3,000 ppm females. At 2 years, the incidences of follicular cell hyperplasia in all exposed groups of mice were significantly greater than those in the controls, but there were no increases in thyroid gland follicular cell neoplasms. The incidence of centrilobular fatty change in the liver of 3,000 ppm males was significantly lower than that in the controls at 15 months, and the decreased incidence appeared to be related to exposure level. At 2 years, the incidences of eosinophilic foci, foci of fatty change, centrilobular cytomegaly, and centrilobular fatty change in 3,000 ppm males were lower than those in the controls. The incidence of hepatocellular adenomas and the incidence of hepatocellular adenomas or carcinomas (combined) in 3,000 ppm males and females were significantly lower than those in the controls; this was considered to be related to lower body weights in these groups. GENETIC TOXICOLOGY: Codeine phosphate was not mutagenic in any of four strains of Salmonella typhimurium, with or without S9 metabolic activation enzymes. In cytogenetic tests with cultured Chinese hamster ovary cells, codeine phosphate induced dose-related increases in sister chromatid exchanges, with and without S9, only at concentration levels that caused cell cycle delay. No induction of chromosomal aberrations was noted in cultured Chinese hamster ovary cells treated with codeine phosphate, with or without S9. CONCLUSIONS: Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity of codeine in male or female F344/N rats exposed to 400, 800, or 1,600 ppm. There was no evidence of carcinogenic activity of codeine in male or female B6C3F1 mice exposed to 750, 1,500, or 3,000 ppm. Thyroid gland follicular cell hyperplasia was increased in exposed male and female mice. Decreased incidences of benign pheochromocytomas of the adrenal medulla in male rats and mammary gland fibroadenomas and fibroadenomas or adenocarcinomas (combined) in female rats were related to codeine exposure. Synonyms: 7,8-didehydro-4,5-epoxy-3-methoxy-17 methylmorphinan-6-ol; methylmorphine; 3-0-methylmorphine monohydrate; N methylnorcodeine; morphine-3-methyl ether; morphine monomethyl ether Trade names: Codeinum, Codicept, Coducept, Metilmorfina PMID- 12587022 TI - Genesis of biochemistry: a problems approach. PMID- 12587024 TI - Approaches in cell biology teaching. PMID- 12587023 TI - Analyzing defects in the Caenorhabditis elegans nervous system using organismal and cell biological approaches. AB - The goal of this laboratory exercise is to increase student understanding of the impact of nervous system function at both the organismal and cellular levels. This inquiry-based exercise is designed for an undergraduate course examining principles of cell biology. After observing the movement of Caenorhabditis elegans with defects in their nervous system, students examine the structure of the nervous system to categorize the type of defect. They distinguish between defects in synaptic vesicle transport and defects in synaptic vesicle fusion with membranes. The synaptic vesicles are tagged with green fluorescent protein (GFP), simplifying cellular analysis. The expected outcome of this experiment is that students will better understand the concepts of vesicle transport, neurotransmitter release, GFP, and the relation between the nervous system and behavior. PMID- 12587025 TI - HHMI: educating tomorrow's scientists. PMID- 12587026 TI - WWW. Cell Biology Education. PMID- 12587028 TI - Report from the ASCB Education Committee. PMID- 12587029 TI - Video views and reviews. PMID- 12587030 TI - Assessing student learning. AB - Biology education research has now reached a level of maturity where the expectation is that researchers will assess the effectiveness of their innovation on student learning. This may include an examination of affective outcomes, such as student attitudes and beliefs, as well as student understanding of discipline based content. A variety of tools are available to generate assessment data, each with certain advantages and disadvantages. They include not only quantitative measures, which lend themselves to familiar statistical analyses, but also qualitative techniques that can provide a rich understanding of complex outcomes. This article describes some of the most commonly used assessment techniques, their advantages and disadvantages, and typical ways such information is reported. PMID- 12587031 TI - Biotechnology apprenticeship for secondary-level students: teaching advanced cell culture techniques for research. AB - The purpose of this article is to discuss small-group apprenticeships (SGAs) as a method to instruct cell culture techniques to high school participants. The study aimed to teach cell culture practices and to introduce advanced imaging techniques to solve various biomedical engineering problems. Participants designed and completed experiments using both flow cytometry and laser scanning cytometry during the 1-month summer apprenticeship. In addition to effectively and efficiently teaching cell biology laboratory techniques, this course design provided an opportunity for research training, career exploration, and mentoring. Students participated in active research projects, working with a skilled interdisciplinary team of researchers in a large research institution with access to state-of-the-art instrumentation. The instructors, composed of graduate students, laboratory managers, and principal investigators, worked well together to present a real and worthwhile research experience. The students enjoyed learning cell culture techniques while contributing to active research projects. The institution's researchers were equally enthusiastic to instruct and serve as mentors. In this article, we clarify and illuminate the value of small-group laboratory apprenticeships to the institution and the students by presenting the results and experiences of seven middle and high school participants and their instructors. PMID- 12587034 TI - [Desfluoroquinolones vs. fluoroquinolones]. PMID- 12587035 TI - [The bacteriophages and their gene products as antimicrobial agents]. AB - The viruses that infect bacteria (bacteriophages or phages) were first isolated about 90 years ago. Phages have been fundamental tools in the development of molecular biology. Phages were early hypothesized as therapeutic agents for combatting pathogenic bacteria. However, the discovery and successful use of antibiotics to treat infectious diseases hindered this aim. the development of bacterial resistance to most available drugs has recently led researchers to test the possibilities of using phages as therapeutic agents. We review here recent achievements in this field, taking into consideration former bias in handling phages as well as previous achievements carried out in Eastern Europe where bacteriophages have been employed for decades as an alternative to antibiotics. PMID- 12587032 TI - Using the two-hybrid screen in the classroom laboratory. AB - The National Science Foundation and others have made compelling arguments that research be incorporated into the learning of undergraduates. In response to these arguments, a two-hybrid research project was incorporated into a molecular biology course that contained both a lecture section and a laboratory section. The course was designed around specific goals for educational outcomes, including introducing research to a wide range of students, teaching students experimental design and data analysis, and enhancing understanding of course material. Additional goals included teaching students to search genomic databases, to access scientific articles, and to write a paper in scientific format. Graded events tested these goals, and a student evaluation indicated student perception of the project. According to our analysis of the data, the yeast two-hybrid screen was a success: several novel clones were identified; students met expectations on graded lab reports, the poster session, and the final paper; and evaluations indicated that students had achieved the outlined goals. Students indicated on the evaluations that the research project increased their interest in research and greatly improved understanding of the course material. Finally, several students in the course intend to submit the findings of the research project to an undergraduate research journal. PMID- 12587036 TI - [Quinolones and Streptococcus pneumoniae. Mechanisms of action and resistance]. AB - Streptococcus pneumoniae is considered the most frequent bacterial cause of community-acquired pneumonia, and is involved in a significant number of cases of acute exacerbations of chronic bronchitis, acute otitis, sinusitis, meningitis and other infectious diseases. Fluoroquinolones have been extensively investigated in recent years in the search for new agents that has been prompted by the emergence of resistance in this microorganism. Furthermore, the study of resistance from a molecular biology standpoint has helped in elucidating almost all the biochemical mechanisms of resistance and the routes of dissemination of genetic information between bacteria. This short review is focused on the mechanism of action of quinolones and on the mechanisms responsible for resistance of S. pneumoniae to them, given their clinical and epidemiological relevance. S. pneumoniae is a case apart because bactericidal activity against this microorganism can be produced through gyrase, topoisomerase IV or both, depending on the quinolone structure, which shows that structure has an influence on the success of treatment. Knowledge of the resistance prototype is therefore important so that the appropriate antibiotic therapy can be recommended when indicated. PMID- 12587037 TI - [In vitro activity of moxifloxacin against respiratory pathogens in Latin America]. AB - We compared the in vitro activity of moxifloxacin, levofloxacin and six other antibiotics frequently used in respiratory tract infections, against 1563 Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Haemophilus influenzae and Moraxella catarrhalis clinical isolates from 21 centers in 10 Latin American countries from March 2000 to April 2001. Moxifloxacin was the most active compound against all the species included. Moxifloxacin was 2- to 4-fold more active than levofloxacin against Gram-positive bacteria. This difference was much higher against levofloxacin-resistant S. pneumoniae isolates (MIC: 0.5 mg/l vs 8 mg/l). The activity of moxifloxacin against H. influenzae and M. catarrhalis was similar to levofloxacin; all the isolates were inhibited at < or = 1 mg/l concentrations of moxifloxacin and levofloxacin. PMID- 12587038 TI - [In vitro activity of posaconazole against yeasts isolated in blood cultures]. AB - The in vitro activity of posaconazole against Candida species isolated from blood cultures and the influence of incubation time was studied and compared with that of fluconazole. A total of 112 isolates were studied: 32 Candida albicans, 33 C. parapsilosis, 17 C. tropicalis, 8 C. glabrata, 8 C. guilliermondii, 3 C. famata, 2 C. lusitaniae, 1 C. lipolytica, 1 C. inconspicua, 1 C. lambica, 3 Saccharomyces cerevisiae, 1 Blastoschizomyces capitatus, 1 Geotricum spp. and 1 Pichia omheri. The MIC was obtained using the M27-A microdilution method described by the NCCLS for Candida spp. and Cryptococcus neoformans. The species most susceptible to posaconazole were C. parapsilosis (MCI90 0.016 mg/l), C. guilliermondii (MIC90 0.12 mg/l), C. glabrata (MCI90 0.5 mg/l) and Candida spp. (MCI90 0.25 mg/l). However, this azole did not improve the activity of fluconazole against C. tropicalis (MIC90 8 mg/l) and C. albicans (MCI90 mg/l). The time of reading was important in the detection of resistance, as the number of strains resistant to fluconazole or posaconazole was higher at 48 hours than at 24 hours for C. albicans and C. tropicalis. All the other species of Candida were susceptible at both reading times. PMID- 12587039 TI - [In vitro activity of clarithromycin and metronidazole against Helicobacter pylori in different incubation atmospheres]. AB - The aim of this study was to determine the in vitro activity of clarithromycin and metronidazole using an agar dilution method to compare two different incubation atmospheres: a CO2 incubator and a jar with a microaerobic gas generating system. Antibiotics were placed on plates in twofold dilutions ranging from 128 to 0.064 mg/l in Mueller-Hinton agar supplemented with 7% horse blood. The inoculum was prepared from 31 Helicobacter pylori isolates and was inoculated using a Steers replicator. Plates were incubated for 3 to 5 days and MICs were recorded as the lowest concentration of antibiotic inhibiting visible growth. Two different incubation atmospheres were used: a CO2 incubator set at 95% humidity and 10% CO2, and a jar with a gas-generating envelope that produces 7-10% O2 and 14% CO2 (BioMerieux). Clarithromycin resistance was found in 19% of strains both in the gas-generating system and the CO2 incubator. Metronidazole resistance was 23% in both atmospheres. MICs for clarithromycin in both atmospheres showed two dilutions of difference for 100% of the strains, and were slightly higher in the jar with a gas-generating envelope. However, MICs for metronidazole were higher when it was incubated in the CO2 incubator, and in 86.7% of strains the MICs showed < or = 2 dilutions of difference. No great discrepancies were found for either metronidazole or clarithromycin using the two methods. PMID- 12587040 TI - [Study of the diversity in treatment of community-acquired pneumonia]. AB - There are a great number of protocols or recommendations focused on clinical radiological criteria for the antibiotic treatment of community-acquired pneumonia (CAP). This report assesses the diversity found in the treatment of CAP in the Internal Medicine and Pneumology Services of our hospital. Seventy-one patients referred from the Emergency Department were treated in both services, and clinical, microbiological and therapeutic variables were collected. It was shown that there are significant differences between the therapeutic guidelines used in each service (antibiotics chosen, their length and form of administration) which could not be explained by the patients' clinical profile. Greater homogeneity in the guidelines was found only for patients with chronic obstructive pulmonary disease. In relation to the adherence to recommendations of scientific societies, we found that 7% of the guidelines were not recommended; there was an average of 2.5 changes (in the antibiotic, dose or administration form) per patient; and a larger period of intravenous administration (7.38 days) than recommended for sequential therapy PMID- 12587041 TI - [Influence of ion pump-inhibiting drugs on the accumulation of ofloxacin and grepafloxacin in human polymorphonuclear leukocytes]. AB - In this study we tested the influence of three ion pump-inhibiting drugs (digoxin, omeprazole and verapamil) on the accumulation of ofloxacin and grepafloxacin in human polymorphonuclear leukocytes. Two assay conditions were established: cell preincubation with the drug for 30 or 60 minutes before addition of quinolone, or addition of both drugs simultaneously. The maximum I/E for ofloxacin is different depending on the assay conditions: 7.69+/-0.88; 5.64+/ 1.91 and 3.56+/-1.04 for the assay without preincubation and with preincubation for 30 or 60 minutes at 37 masculine C, respectively. Similarly, grepafloxacin reached the following maximums: 61.27+/-3.04; 32.18+/-3.25 and 22.52+/-3.86. Digoxin did not significantly modify the accumulation of the quinolones, but it increased the I/E compared with the control. In general, omeprazole reduced the accumulation of both quinolones. When omeprazole and ofloxacin were added together, ofloxacin's I/E was significantly lower; however, for grepafloxacin, 60 minutes of preincubation were necessary. Verapamil induced a significant increase in the I/E for both quinolones when the cells were preincubated at 10 times the plasma concentration. PMID- 12587042 TI - [Pseudomonas aeruginosa bacteremia: clinical and epidemiological patterns and treatment]. AB - Pseudomonas aeruginosa bacteremia has the highest mortality among Gram-negative infections. We studied its clinical and epidemiological patterns in order to achieve an early and accurate diagnosis and treatment. We prospectively studied 211 cases of P. aeruginosa bacteremia admitted to our hospital from 1991 to 1998. All patients had at least one positive blood culture for P. aeruginosa and clinical symptoms of bacteremia. The prognosis of the underlying disease, past illnesses, foci location, initial clinical status, type of bacteremia, complications, and the type of antibiotic treatment were analyzed. Clinical and microbiological controls were carried out at baseline, at the end of the treatment period, and 1 month later. Of the 211 cases, 73% were men with a mean age of 56.5 years. Overall, 45.5% were hospitalized in the ICU, 28% in the surgical ward, and 26% in the medical service, with a mean length of stay of 40.4 days. As risk factors we identified a length of stay more than 14 days (76.7%), previous manipulations (87.6%), and surgery (44.5%). A total of 43.1% had had an infection in the last 6 weeks, and 46.4% were treated with antibiotics, usually broad-spectrum antibiotics without activity against P. aeruginosa. The most frequent underlying diseases were neoplasia (25.5%), postsurgical cardiopathy (24.1%), nephrosis (22.7%), chronic obstructive pulmonary disease (17.5%) and diabetes (13.2%). The most frequent foci were pulmonary (29.3%), urinary (21.8%), and undetermined (38.3%). The mortality rate was 27.9% and was considered related to P. aeruginosa bacteremia in 76.2% of the cases. The antibiotic treatment was considered adequate in 89% of the cases. It was concluded that knowledge of the clinical and epidemiological factors of patients with P. aeruginosa bacteremia may help to optimize its management and to reduce its morbidity and mortality. PMID- 12587043 TI - [ROC curve analysis of factors predictive of response to treatment with interferon plus ribavirin in patients with chronic hepatitis C relapse after previous interferon treatment]. AB - This study aimed to identify the factors predictive of response before the initiation of treatment and throughout the treatment period in patients with chronic hepatitis C relapse after treatment with interferon-a who were retreated with a standard regimen of interferon-a plus ribavirine and followed up for 40 months. Forty-four patients (40 with genotype 1, four without genotype 1) were included in the study. Four patients (genotype 1) were excluded because of adverse effects. The rate of maintained response was 55% (50% genotype 1, 100% non-genotype 1). The stage of histological damage (>2), glutamic-pyruvic transaminase (GPT) concentration (< or = 26 UI/l) and the association between the GPT concentration and the detection of the RNA-HCV in the first and third treatment months were the variables with an area under the ROC curve and a confidence interval >0.5. The probability of predicting a maintained response (negative predictive value) if the stage of histological lesion was <2 was 62.9%, while the positive predictive value was 100%. During the treatment, the disappearance of the RNA-HCV together with GPT values < or =26 in the first treatment month were the best predictive values. In this case, the negative predictive value was 78.3% and the positive predictive value was 76.5% (OR: 11.7, 2.6-52.2). Furthermore, the GPT value with the best predictive value (<26 UI/l) was a more effective predictor of the response to treatment than the normal value of the GPT. Finally, the GPT values >26 UI/l and the detection of RNA/HCV in the first or third treatment month were certain predictors of the absence of response but with low sensitivity (10-12%). It was concluded that is possible to predict the response to the combined treatment with an acceptable level of confidence, although not unequivocally. Ninety percent of the patients would be candidates for maintaining treatment for at least 6-12 months, while approximately 10% could undergo early interruption of treatment due to the absence of response. PMID- 12587044 TI - Who is WHO and what was REAL? AB - The principles of the new WHO classification of haematopoietic and lymphoid tumours are based on those defined in the Revised European American classification of Lymphoid neoplasms (REAL), published by the International Lymphoma Study Group (ILSG) in 1994. Thus, the new WHO classification may be considered an updated version of the REAL classification rather than of the old WHO classification published in 1976. Disease entities are defined on the basis of morphological, phenotypic, genotypic, and clinical data. The relative impact of these characteristics varies among different diseases and there is "no gold standard". Thus, the strict hierarchy among diagnostic criteria, headed by morphology and followed by immunohistochemistry and genetics, has been discontinued. The WHO classification not only encompasses lymphoid tumours but extends to myeloid, mast cell and histiocytic/dendritic cell malignancies. Neoplasms are primarily stratified according to their tumour cell lineage. For each neoplasm a cell of origin is postulated. The classification of lymphoid malignancies recognises three major categories, B-cell neoplasms, T-/NK-cell neoplasms, and Hodgkin lymphomas. B-cell and T-cell lymphomas are further divided into precursor neoplasms and mature neoplasms, the latter being subdivided according to their clinical manifestation into disseminated/leukaemic, extranodal and nodal malignancies. In contrast to previous classifications, the neoplasms are grouped neither according to their histological grade (Kiel classification) nor according to their clinical aggressiveness (International Working Formulation). However, the histological grade is considered a prognostic factor which enters into the description of each disease entity. Hodgkin's disease, now more appropriately termed Hodgkin lymphoma, comprises nodular lymphocyte predominant Hodgkin lymphoma and classical Hodgkin lymphomas of nodular sclerosis, mixed cellularity, lymphocyte-depleted and lymphocyte-rich subtype. For practical purposes this minireview disregards the description of myeloid, macrophage/histiocytic, dendritic cell and mast cell disorders. Furthermore, the present paper is restricted to those lymphoid tumours that are not already identically described in the REAL classification, in order to focus on what is really new in the WHO classification. PMID- 12587045 TI - B-type natriuretic peptide (BNP): can it improve our management of patients with congestive heart failure? AB - Until recently no simple specific test existed for the differentiation of decompensated heart failure from other causes of acute dyspnoea, or to assess the prognosis of patients with severe heart failure or to optimize heart failure therapy in an individual patient. Measurement of B-type natriuretic peptide has become available as an easy-to-per-form bedside test. Several studies have demonstrated it's usefulness in the emergency room to differentiate heart failure from other causes of acute dyspnoea or to guide the complex drug therapy in an individual patient with heart failure. This article gives a short overview on the clinical experience to use BNP-blood levels for the diagnosis and treatment guidance of heart failure. PMID- 12587046 TI - B-type natriuretic peptide for diagnosis and treatment of congestive heart failure. AB - BACKGROUND: Prognostic classification of congestive heart failure (CHF) is difficult and only possible with the help of additional diagnostic tools. B-type natriuretic peptide (BNP) has been used as a diagnostic and prognostic marker for patients (pts) with CHF. In this study, the clinical value of BNP for stratification and treatment of pts with CHF was evaluated. PATIENTS AND METHODS: 33 out-pts with CHF (age 57 +/- 12 years) were included. Left-ventricular (LV) ejection fraction (EF) was 27 +/- 8% (mean +/- SD) and NYHA-class 2.4 +/- 0.7. Following parameters were measured: BNP and sodium from blood samples, exercise performance from 6-minute walking test (6MWT, meters) (n = 18), LV end-diastolic diameter (LVEDD) and LV mass (LVM) from 2D-echocardiography (n = 33), as well as LV end-diastolic pressure (LVEDP, n = 23) and systemic vascular resistance (SVR, n = 20) from heart-catheterisation. Ten pts were hospitalised in the preceding 6 months because of worsening CHF or for optimisation of medical therapy. BNP was measured at the beginning and end of the hospital-stay. Follow-up was for 1 year. RESULTS: Pts with a high NYHA-class had a higher BNP (pg/ml) than those with a low NYHA- class: NYHA I 51 +/- 20, II 281 +/- 223, III 562+/-346 and IV 1061 +/- 126 pg/ml (p = 0.002). BNP correlated with LVEDP (r = 0.50, p <0.02), SVR (r =0.49, p <0.03) and inversely with 6MWT (r =-0.60, p <0.009), LVEF (r = -0.49, p <0.004) and sodium (r = -0.36, p = 0.04). In the hospitalised pts, mean BNP (pg/ml) was 881 +/- 695 at admission,and 532 +/- 435 at discharge (n.s.). Decrease in BNPduring hospitalisation paralleled weight-loss and was significantly greater in patients with >1000 pg/ml BNP at admission (n = 5) as compared to the 5 patients with BNP <1000 (p <0.03). Patients with an adverse event during 1-year follow-up had significantly higher BNP both at steady-state (603 +/-359 pg/ml) and at time of decompensation than patients with a favourable outcome (227 +/- 218 pg/ml,p <0.001). CONCLUSIONS: BNP correlates well with the clinical severity of CHF (NYHA-class) and is directly related to filling pressure (LVEDP), LV function(LVEF) and exercise performance (6 MWT). Furthermore, BNP has prognostic impact with regard to adverse clinical events. PMID- 12587047 TI - Validity of self-reported exercise-induced sweating as a measure of physical activity among patients with coronary artery disease. AB - BACKGROUND: In several epidemiological studies, self-reported exercise-induced sweating has been used as a measure of physical activity (PA). Among healthy subjects it is moderately well associated with other measures of PA and physical fitness, but its validity among patients with coronary artery disease (CAD) has not yet been established. METHODS: In a self-administered questionnaire, 233 patients undergoing coronary angiography replied to the question 'In the past 4 weeks, how often have you exercised vigorously enough to work up a sweat?" Patients also gave the frequency and duration of different PA. We examined the association of the sweat frequency question with light (<4 metabolic equivalents [METs]), moderate (4.0-5.9 METs) and intense (>6 METs) PA, and tested the association with maximum exercise capacity. RESULTS: The frequency of self reported exercise-induced sweating was significantly (p <0.001) associated with both moderate and intense PA. The correlation coefficient of moderate and intense PA with sweat frequency was r = 0.34 (p<0.001), and r = 0.44 (p <0.001) for the respective PA of >30 minutes' duration. The strength of the association between sweat frequency and caloric expenditure per time was similar (moderate and intense PA, r = 0.43, p <0.001). There was a significant correlation between maximum exercise capacity measured in METs and sweat frequency (r = 0.37, p <0.001). CONCLUSIONS: These results suggest that for studies among patients with CAD, assessment of the frequency of self-reported exercise-induced sweating provides useful information regarding moderate and intense PA as well as physical exercise capacity. PMID- 12587048 TI - Penatrating cardiac injuries. Complex injuries and difficult challenges. PMID- 12587049 TI - [The effect of heterologous collagen in neochondrogenesis from free perichondrial grafts, in rabbits: an experimental study]. AB - BACKGROUND: After chondrogenic potential of perichondrium was recognized, perichondrial grafts have been used in reconstruction of cartilage defects. Enhancing neochondrogenesis has many advantages, in this regard. This experimental study was done in the aim of identifying whether the heterologous collagen has an accelerating effect on neochondrogenesis from perichondrial grafts or not. METHODS: We placed perichondrial grafts, onto the dorsal muscles of twelve white New-Zealand rabbits, on both sides. On one side, heterologous collagen sponge was inserted between the graft and the muscle, the other side served as a control. RESULTS: As a result of histological assessment on days 45, 120 and 180, we obtained that heterologous collagen enhanced the neochondrogenesis. CONCLUSION: We believe that, if any cartilage defect would be reconstructed with a Perichondrial graft, additional use of heterologous collagen will be helpful. PMID- 12587050 TI - [The comparison of the results of the conservative treatment between isolated solid organ injuries and those injuries associated with extraabdominal injuries after blunt abdominal trauma between isolated solid organ injuries and those injuries associated with extraabdominal injuries after blunt abdominal trauma]. AB - BACKGROUND: To determine whether conservative treatment in blunt abdominal solid organ injuries associated with extraabdominal trauma is safe or not. METHODS: Between December 1998 and December 2001, 49 patients were admitted with blunt abdominal trauma, 34 of whom were hemodynamically stable. These 34 patients were divided into two groups. Seventeen patients had isolated abdominal solid organ injuries (Group I) and 17 patients had associated extraabdominal injuries (Group II). The results of conservative treatment, blood pressure values, pulse rates, ages, gender, abdominal solid organ injury scores, the etiologies of trauma, the number of the blood transfusions, morbidity and mortality rates, and hospital stays were compared. RESULTS: The blood pressure values and pulse rates as measured in our emergency room, abdominal solid organ injury scores, ages, gender, the results of conservative treatment, the etiologies of trauma, morbidity and mortality rates were similar in both groups. Comparisons between hospital stays and numbers of the blood transfusion have show statistically significant differences in favour of group II. No patients had hollow viscus injury. CONCLUSION: Conservative treatment is safe for hemodynamically stable blunt abdominal trauma patients with solid organ injury but no hollow viscus injury, even if they have extraabdominal injuries. The existence of extraabdominal injuries, however, prolongs the hospital stay and increases the need of the blood transfusion. PMID- 12587051 TI - [Cardiac penetration wounds: three years Adana Numune Hospital experience]. AB - BACKGROUND: Cardiac penetration wounds have high mortality which need a rapid diagnosis and surgical intervention. METHODS: Between May 1999 to May 2002, 196 thoracic penetration cases who were admitted to emergency department were examined and in 12 (6.1%) of them cardiac penetration was diagnosed. In this retrospective study, the cardiac penetration cases were evaluated. RESULTS: All the cases were male, 11 of them were injured by stab wounds, and one case by a gunshot wound. We performed left anterolateral thoracotomy in nine of the cases, right anterolateral thoracotomy in two cases and a median sternotomy in one case. Two of the cases died. CONCLUSION: The survival chance of the cases can be increased with rapid diagnosis and surgical intervention. PMID- 12587052 TI - [Evaluation of severe burns managed in intensive care unit]. AB - BACKGROUND: This study designed to evaluate the results of the patients with severe burns treated in the intensive care unit. METHODS: Between May 1997 and May 2002, fifty-four patients who had thermal and electrical burns managed at the Intensive Care Unit of General Surgery of Istanbul University Medical Faculty. RESULTS: Forty-five of patients were males and nine were females with mean age of 39 years. The mean hospital stay was 26 days. The mean total body surface area burns was 60%. Pseudomonas aeruginosa was detected in wound cultures at the 68% of patients. 83% of cases died. Mortality was due to sepsis in 66% of patients. CONCLUSION: Besides early debridmant, early enteral feeding, woundcare and intensive care unit support, establishing of specific burncenters may reduce morbidity and mortality rates in severe burns. PMID- 12587053 TI - [Venous thromboembolism prophylaxis with low molecular weight heparins in polytraumatized patients in intensive care unit (extended serie)]. AB - BACKGROUND: In our study we aimed to evaluate, retrospectively, the bleeding and the Venousthrombo Emblism (VTE) complications in trauma patients in Intensive Care Unit (ICU) under the prophylaxis of enoxaparin sodium. The results will guide us to establish a protocol for use of Enoxaparin Sodium in trauma patients. In trauma patients, Low Molecular Weight Heparine (LMWH) has better efficiency than unfractionned heparin and intermittant pneumatic compression for VTE prophylaxis. METHODS: 457 polytraumatized patients treated in ICU are included to the study group. All patients received 40 mg/day of Enoxaparin Sodium. Severity of trauma was assessed with Apache II Scoring System and bleeding diagnosed by observing a sudden drop of 2 g/dl in hemoglobin concentration and pulmonary embolism was diagnosed by a sudden change in blood gases and deterioration of the clinical outlook which was confirmed by a spiral CT scan. Statistical correlation was made by Pearson's correlation test. RESULTS: Mean Apache II score was 13.8 with a total mortality of 41%. 42 patients (9,2%) had bleeding due to Enoxiparine Sodium prophylaxis. 12 patients (2,6%) had pulmonary embolism and 8 of them (1,7%) had died. CONCLUSION: Verified by the literature and our study, LMWH prophylaxis significantly reduces the Pulmoner Embolism (PE) incidence in polytraumatized patients (p<0,05). On the other hand, the bleeding risk has slightly increased without showing any significance (p>0.05). Prophylaxis with LMWH is safe and efficient in polytraumatized ICU patients. PMID- 12587054 TI - Foreign body aspiration: a four-years experience. AB - BACKGROUND: Foreign bodies (FB) in the airway require prompt removal in children. We reviewed our experience in patients with suspected airway FB. METHODS: A retrospective study was conducted in 189 consecutive children who admitted to the Department of Pediatric Surgery, SSK Tepecik Training Hospital between 1997-2001. Patients data on presentation, bronchoscopy findings and results were obtained. Of the 189 bronchoscopies, 127 (67.2%) showed FB which are commonly located in the right mainstem bronchus. Most of FB were nonradiopaque. Pips and hazelnuts were the most common FB. Mean hospital stay was 2.5 days. Many patients (151 out of 189; 79.9%) had transient stridor or fever that ceased within 24 hours after bronchoscopy. No mortality was observed rin relation with bronchoscopy. CONCLUSION: Chest radiographs of the children with FB in the airways are inconclusive. Children with a history of small particles in their mouths and subsequently showing wheezing, or choking episode should undergo prompt bronchoscopy. Complications related to bronchoscopy are uncommon. PMID- 12587055 TI - [Intraabdominal and intramural hemorrhage due to warfarin therapy]. AB - BACKGROUND: The study investigated clinical, radiological findings and treatment methods used in patients who developed intraabdominal hemorrhage due to chronic warfarin use. PATIENTS AND METHODS: Eight patients receiving warfarin for cardiac valve replacement, pulmonary thromboemboli, and atrial fibrillation were admitted to our hospital. The patients had abdominal pain, nausea, and vomiting although there was no hematemesis and melena. RESULTS: Abdominal ultrasonography and computerized tomography identified the individuals' problems as intraabdominal hemorrhage (n=2), intestinal intramural and intraabdominal hemorrhage (n=2), bleeding into the sheath of the rectus abdominus muscle (n=1), subcapsular splenic hemorrhage (n=1), and bleeding due to ruptured ovarian cyst (n=2). CONCLUSIONS: Clinicians must be alert for intraabdominal bleeding in patients who are prescribed warfarin treatment. Abdominal ultrasonography and computerized tomography should be used to investigate all such cases of suspected hemorrhage. Blood and plasma replacements are first line of supportive treatment and surgery should be avoided if possible. PMID- 12587056 TI - [Surgical treatment of penil fractures]. AB - BACKGROUND: Patients diagnosis as penil fracture and treated with surgical methods were evaluated retrospectively between January 1990-February 2002. METHODS: Patients were evaluated by age, trauma type, time passed after the trauma, physical examination and radiologic data. RESULTS: The cause of trauma was blunt trauma in all patients. 14 trauma (60.8%) were during sexual activity, 6 trauma (26%) were during handling the erective penis in morning erection, 3 of trauma (13%) were due to the rolling on to the penis. Urethral bleeding was seen in 1 patient and microscopic hematuria was detected in 5 patients. Subcoronal circular incision was carried out in 16 patients (69.5%), semicircular incision was done directly on the injury in 6 patients (26%) and from penis radix to scrotum was done in 1 patient (4.3%). There was unilateral corpus cavernosum injury in 21 patients. Bilateral corpus cavernosum injury in 1 patient, corpuscavernosum, corpus spongiosum and incomplent urethral injury in 1 patient. Wound infection was detected only in 2 patients at postoperatively early period. After evaluating 20 patients with medical history and physical examination in postoperative control penil curvature permitting sexual activity was detected in 3 patients. CONCLUSION: We concluded that patients with penil fracture may be treated with low complication rate by early surgical procedure. PMID- 12587057 TI - [Anterior dural laseration due to thoracolumbar burst fracture]. AB - BACKGROUND: This study investigated the incidence, prediction, and treatment principles of anterior dural laseration due to thoracolumbar burst fracture. METHODS: Three anterior dural tears were found during anterior decompressive spine surgery. The average age was 42. Preoperative neurologic status and radiologic data of the patients were analyzed retrospectively. The average follow up was 20 months. RESULTS: Anterior dural tears detected in 3 (8.2%) of 35 burst fractures. Primary repair were performed in two patients. There were no cerebro spinal fluid leakage perioperatively and all patients were neurologically intact at the final follow-up. CONCLUSION: It is too difficult to detect anterior dural tear before surgery. Anterior dural tear should be suspected in patient with neurologic deficit and asimetric bony retropulsion into the spinal canal. PMID- 12587058 TI - [The treatment of displaced supracondylar fractures of the humerus in children (an evaluation of three different treatment methods)]. AB - BACKGROUND: Supracondylar fractures of the humerus are the most common fracture pattern of the elbow in childhood. They require close follow-up subsequent to successful treatment, because of having a great risk of many complications. Many treatment methods have been described for the treatment of displaced supracondylar fractures of the humerus, however it has not been reached a consensus for the choice of the treatment. The purpose of this study is to investigate the late results of three different treatment methods; closed reduction+cast immobilization, skeletal traction+cast immobilization and open reduction+internal fixation by K-wires. We also aimed to compare these methods with the results of current literature. METHODS: Fifty-three (41 boys and 12 girls) displaced supracondylar humerus fractures were treated between January 1986 and March 1999. The average age was 8.0 years. We evaluated the results of patients according to Flynn's criterias. RESULTS: According to the results of the study, we obtained 31 excellent, 9 good, 5 fair and 8 poor results. The ratio of poor results of closed reduction+cast immobilization, skeletal traction+cast immobilization and open reduction+internal fixation by K-wires were 28.6%, 14.3% and 12.8%, respectively. CONCLUSION: The datas in our study show that closed reduction+cast immobilization results in the most poorest outcome in the treatment of displaced supracondylar humerus fractures. PMID- 12587059 TI - Post-traumatic testicular torsion. AB - Torsion of the testis is the most common urological emergency and should be included in the differential diagnosis of scrotal trauma. Clinical suspicion of testicular torsion is a serious indication for prompt surgical exploration of post-traumatic acute scrotum. A case of post-traumatic testicular torsion, which resulted in orchiectomy, is presented and literature is reviewed. PMID- 12587060 TI - [Traumatic dissection of bilateral vertebral arteries]. AB - 58-year-old male comatose patient following a motor vehicle accident with a history of coronary bypass surgery 5 years ago has been presented. The patient was awake after the accident and he became comatose 30 minutes after the accident. The early diagnostic investigations showed no responsible cranial or spinal lesion. At the 2nd day of the accident, diffusion weighted magnetic resonance scans, magnetic resonance arteriography and digital subtraction arteriography revealed bilateral vertebral artery dissection at the second part of the vertebral arteries. Clinical, radiological features and treatment options of the vertebral artery dissection has been discussed with the ertinent literature. PMID- 12587061 TI - Chance type cervical fracture and neurological deficits in ankylosing spondylitis. AB - Prevention of sudden neck movements is vital in patients with ankylosing spondylitis of the cervical spine. We present a case of ankylosing spondylitis who sustained a cervical fracture. He presented with paraplegia after a minor car collision and died of pulmonary embolism after the operation for anterior stabilisation. We believe that the most important matter in a patient with advanced ankylosing spondylitis is the prevention of the fractures and complications. The need for neck protection in automobiles was emphasized and the literature reviewed about the occurrences of neurological deficits following trauma. PMID- 12587062 TI - [Posttraumatic tension pneumocephalus causing herniation]. AB - A 12 year old boy was admitted to emergency service with loss of consciousness, convulsions and anisocoria of the right eye along the history of upper respiratory tract infection for the last four days. Two months ago he was hospitalized at the intensive care unite due to motorcycle accident with frontal fracture and minimal frontal pneumocephalus. He was treated for ten days and discharged without any neurological impairment. The last computerized tomography showed right frontal air of 5x5x7 cm. multiple airs in the ventricular system and in the upper central canal of the cervical region. The air was removed immediately through a right frontal burr-hole with a bedside procedure. The patient's neurological status rapidly improved and ten days after computerized tomography showed clearly resolution of the air. The presented case is an interesting example of post traumatic pneumocephalus due to herniation and widespread intracranial air. The situation can be explained with acute penetration of huge amount of air into the cranium through the frontobasal fracture and torn dura by strong Valsalva 's maneuver resulting from coughing and sneezing in the course of upper respiratory tract infection. We suggest close follow up, frequent control computerized tomography scans, preventive and effective treatment respiratory tract infections along 3-6 months in similar cases. PMID- 12587063 TI - [Malign melanoma of the small bowel as a cause of occult intestinal bleeding: case report]. AB - Although the metastases from extra- abdominal tumors to the small bowel are rare, cutaneous melanoma is the most common source. A 78 year old man with occult intestinal bleeding due to malign melanoma of the small bowel was admitted to our clinics. The patient was managed successfully by small bowel resection with end to end anastomosis. Patient is alive three months after the surgery and is in pretty good condition. PMID- 12587064 TI - A novel neurotropic expression vector based on the avirulent A7(74) strain of Semliki Forest virus. AB - Semliki Forest virus (SFV), an enveloped alphavirus of the family Togaviridae, infects a wide range of mammalian host cells. Most strains are neurotropic but differ in virulence. The authors took advantage of the nonpathogenic properties of SFV strain A7(74), cloned recently in their laboratory, and constructed a replication-proficient expression vector to target the central nervous system (CNS) for heterologous gene expression. The vector, termed VA7, was engineered to drive expression of foreign inserts through a second subgenomic promoter inserted in the viral 3' nontranslated region (NTR). Infectious virus was obtained by in vitro transcription and transfection into BHK cells, and was shown to direct synthesis of heterologous proteins in several mammalian cell lines. Although novel expression vehicle is not applicable for targeting specific cell populations within the CNS in its present form, in cultured rat hippocampal slices, VA7 encoding enhanced green fluorescent protein (EGFP) efficiently transduced pyramidal cells, interneurons, and glial cells. With prolonged time post infection, the number of EGFP-expressing neurons in hippocampal slices increased. Mice infected intraperitoneally with the recombinant virus remained completely asymptomatic but showed CNS expression of EGFP as evidenced by immunohistochemistry. SFV A7(74) is a nonintegrating virus, which gives rise to a randomly distributed, patchy infection of the adult CNS that is cleared within 10 days. With the advantage of noninvasive administration, the expression vector described in this work is thus applicable for short-term gene expression in the CNS. PMID- 12587065 TI - Semliki Forest virus A7(74) transduces hippocampal neurons and glial cells in a temperature-dependent dual manner. AB - In central nervous system (CNS) tissue preparations, wild-type Semliki Forest virus (SFV) mainly infects neurons, and in vivo it causes lethal encephalitis in neonatal and adult rodents. The SFV strain A7(74), by contrast, is avirulent in adult rodents, triggering only limited CNS infection. To examine A7(74) infection in hippocampal tissue, the authors constructed a replicon, termed SFV(A774nsP) GFP, expressing green fluorescent protein. The results were compared to replication-proficient recombinant A7(74) encoding GFP, named VA7-EGFP. As nonstructural gene mutations can confer temperature sensitivity, the authors also tested whether infection was temperature-dependent. Indeed, at 31 degrees C both viral recombinants transduced significantly more baby hamster kidney cells than at 37 degrees C. When rat hippocampal slices and dissociated cells were incubated at 37 degrees C, SFV(A774nsP)-GFP transduced glial cells but virtually no neurons the opposite of conventional SFV. For VA7-EGFP at 37 degrees C, the preferred GFP positive cells in hippocampal slices were also non-neuronal cells. At 31 degrees C, however, a more wild-type phenotype was found, with 33% and 94% of the GFP positive cells being neurons for SFV(A774nsP)-GFP in slices and dissociated cells, respectively, and 94% neurons for VA7-EGFP in slices. Immunochemical and electrophysiological analyses confirmed that at 37 degrees C virtually all cells transduced by SFV(A774nsP)-GFP in slices were astrocytes, while at 31 degrees C they also contained neurons. These results show that in addition to the developmental age, the temperature determines which cell type becomes infected by A7(74). Our data suggest that A7(74) is avirulent in adult animals because it does not readily replicate in mature neurons at body temperature, whereas it still does so at lower temperatures. PMID- 12587066 TI - Usefulness of proviral load measurement for monitoring of disease activity in individual patients with human T-lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis. AB - High human T-lymphotropic virus type I (HTLV-I) proviral load in peripheral blood mononuclear cells (PBMCs) has been reported in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and the proviral load has been reported to fluctuate in individual patients during the course of the disease. Clinical symptoms usually became stable after a prolonged period of symptom progression. However, the authors have experienced having some patients whose clinical manifestations suddenly became worse during the course of the disease. To clarify the role of high proviral load and its fluctuation in the pathogenesis of HAM/TSP, the authors measured the proviral load of serially taken PBMCs as well as of cerebrospinal fluid (CSF) cells from patients with HAM/TSP on long term follow-up and compared these with their clinical manifestations. There was a wide distribution of proviral load, from 0.3 to 37.8 copies/100 PBMCs; however, the proviral load in individual patients was relatively stable during the course of the disease. Eighty-three percent of the patients with clinical worsening showed an increase in proviral load at the time point when clinical worsening was recorded, or at the preceding time point. The proviral loads in CSF cells were higher than those in PBMCs in individual patients. The ratio of proviral loads in CSF cells/in PBMCs, but not the absolute load, in either compartment, was significantly associated with clinically progressive disease and with recent onset of HAM/TSP. These findings indicate that clinical progression of HAM/TSP is associated with increased proliferation or immigration of HTLV-I-infected lymphocytes in the central nervous system. PMID- 12587067 TI - CD4+ and CD8+ cells accumulate in the brains of acquired immunodeficiency syndrome patients with human immunodeficiency virus encephalitis. AB - To test the hypothesis that CD4+ T lymphocytes accumulate in brains of end-stage acquired immunodeficiency syndrome (AIDS) patients, we examined T-lymphocyte subsets in the CA1, CA3, and CA4 regions of the hippocampus of AIDS patients with (n = 10) and without (n = 11) human immunodeficiency virus encephalitis (HIVE) plus controls (n = 7). HIV p24 antigen was common in monocytic cells and rare in activated/memory CD45RO+ lymphocytes. Hippocampal activated/memory CD45RO+ T lymphocytes significantly increased (P <.001) in seven of the eight hippocampal subregions with hippocampal HIVE (1.14 +/- 1.4 T cells/high-power field [hpf]), but AIDS hippocampus without HIVE were similar to controls (0.03 +/- 0.07 T cells/hpf and 0.03 +/- 0.09 T cells/hpf, respectively). CD45RO+ and CD3+ lymphocytes were similar in numbers and distribution, whereas CD4+ and CD8+ lymphocytes were weakly immunoreactive and less frequent. All four lymphocyte subtypes were present in perivascular spaces and microglial nodules of HIVE, and had direct contact with neurons. Monocytes, microglia, and multinucleated giant cells were immunoreactive for CD4 in AIDS cases with hippocampal HIVE but microglia in remaining AIDS cases and controls were CD4-. CD68+ macrophages significantly increased in hippocampus of HIVE patients (P <.05) and were predominately perivascular in the absence of local HIVE. These studies show that CD4+ T lymphocytes, as well as CD8+ T lymphocytes, participate in the local inflammatory response of HIVE in end-stage AIDS patients, and suggest that their recruitment requires local HIV infection. The perineuronal location of CD4+ cells provides the potential for lymphocyte-mediated neuronal injury or trans-receptor mediated neuronal infection. PMID- 12587068 TI - Human immunodeficiency virus type 1 gp120 and ethanol coexposure in rat organotypic brain slice cultures: Curtailment of gp120-induced neurotoxicity and neurotoxic mediators by moderate but not high ethanol concentrations. AB - Human immunodeficiency virus type 1 (HIV-1) envelope protein gp120, implicated with other retroviral proteins in acquired immunodeficiency syndrome (AIDS) related dementia, causes neuronal degeneration by inciting cascades of neurotoxic mediators from glia. It also may facilitate neuronal glutamate (N-methyl-D aspartate, NMDA) receptor-mediated excitotoxicity by interacting at the glycine coagonist site. The authors reported that preconditioning rat organotypic hippocampal-cortical slice cultures subchronically with ethanol at concentrations occurring during moderate drinking (20 to 30 mM) prevented gp120's induction of neurotoxic mediators and intracellular calcium, as well as neuronal death. The authors now find that the acute copresence of ethanol in moderate as opposed to high concentrations similarly blocks the retroviral protein's neurotoxic effects in brain slice cultures, assessed with lactate dehydrogenase (LDH) release and propidium iodide (PI) labeling. As with ethanol preconditioning, neuroprotection against gp120 by moderate ethanol coexposure appears secondary to abrogation of the retroviral protein's early induction of arachidonic acid (AA), glutamate, and superoxide (but not nitric oxide) elevations/release. Additionally, experiments indicate that 30 mM ethanol is sufficient to inhibit the NMDA receptor, particularly in the presence of added glycine, thus hindering potential direct neuronal stimulation by gp120. However, in contrast to moderate ethanol, 100 mM ethanol, a concentration tolerated only in chronic alcoholics, potentiates gp120 dependent neurotoxicity (PI labeling) in the hippocampal CA1 region, augments LDH release, and fails to curtail gp120's actions on AA, glutamate, and superoxide but does suppress nitric oxide induction. The results indicate dominant roles for AA, superoxide, and glutamate-mediated oxidative stress in gp120's neurotoxic mechanism, but perhaps a less important role for NMDA receptor stimulation, which would be constrained at both ethanol concentrations employed. We suggest that ethanol's concentration-dependent, two-edged sword behavior could alter the development of dementia in HIV-1-infected individuals during social consumption or abuse. Further studies are needed to elucidate the differing apparently glial effects of the two concentrations of ethanol. PMID- 12587070 TI - Impact of direct virus-induced neuronal dysfunction and immunological damage on the progression of flavivirus (Modoc) encephalitis in a murine model. AB - Flavivirus encephalitis is believed to be the result of two main mechanisms: (i) direct damage to and dysfunction of neurons as a result of viral replication and (ii) destruction of the brain tissue by an inflammatory response. The differential impact of both mechanisms on the progression of flavivirus encephalitis has not been clearly determined. We have now studied the encephalitis caused by Modoc virus (MODV) infection in (i) severe combined immunodeficiency (SCID) mice, (ii) immunocompetent NMRI mice, and (iii) NMRI mice under varying immunosuppressive treatment regimens. In SCID mice, Modoc virus infection proved to be uniformly lethal (100%). The virus replicated extensively in neurons and no signs of inflammation of the brain were observed. In immunocompetent NMRI mice, intranasal (but not intraperitoneal) inoculation with MODV caused severe encephalitis accompanied by a fulminate inflammatory response. When NMRI mice, infected with MODV via the intraperitoneal route, were submitted to a brief immunosuppressive treatment, they also developed encephalitis with an obvious inflammatory component. These animals died significantly earlier than NMRI mice, which received immunosuppressive treatment for a longer period of time. In the latter group, no signs of inflammation of the brain were noted. These models thus allow us to distinguish between the impact of direct viral replication and that of immunological factors in the development of MODV encephalitis, and let us to conclude that (i) replication of the virus in neurons is sufficient to cause fatal encephalitis and (ii) immunological factors contribute significantly to disease progression. PMID- 12587069 TI - Structural and functional evolution of human immunodeficiency virus type 1 long terminal repeat CCAAT/enhancer binding protein sites and their use as molecular markers for central nervous system disease progression. AB - The appearance and progression of human immunodeficiency virus type 1 (HIV-1) associated pathogenesis in the immune and central nervous systems is dependent on the ability of the virus to replicate in these compartments, which is, in turn, controlled by numerous factors, including viral binding and entry, receptor and coreceptor usage, and regulation of viral expression by the long terminal repeat (LTR). The LTR promotes viral expression in conjunction with viral and cellular regulatory proteins, including members of the CCAAT/enhancer binding protein (C/EBP) family, which modulate LTR activity through at least two cis-acting binding sites. Previous studies have shown that these sites are necessary for HIV 1 replication in cells of the monocyte/macrophage lineage, but dispensable in T lymphocytes. To establish potential links between this important family of transcription factors and HIV-1-associated pathogenesis, C/EBP site I and II sequence variation in peripheral blood mononuclear cell (PBMC)-derived LTRs from HIV-1-infected patients with varying degrees of disease severity was examined. A high prevalence of C/EBP site variants 3T (site I) and consensus B (site II) within PBMC-derived HIV-1 LTRs was shown to correlate with late stage disease in HIV-1-infected patients. These results suggest that the increased prevalence in the PBMCs of HIV-1 LTRs containing the 3T C/EBP site I variant and the consensus B site II variant may serve as a molecular marker for disease progression within the immune system. The relative low or high binding affinity of C/EBP beta to sites I and II in electrophoretic mobility shift (EMS) analyses correlated with low or high LTR activity, respectively, in transient expression analyses during both early and late disease stages. The 3T C/EBP site I was the only variant examined that was not found in LTRs derived from PBMCs of patients at early stages of HIV-1 disease, but was found at increasing frequencies in patients with late stage disease. Furthermore, the 3T C/EBP site I was not found in brain derived LTRs of patients without HIV-1-associated dementia (HIVD), but was found in increasing numbers in brain-derived LTRs from patients diagnosed with HIVD. The C/EBP site I 3T variant appears to be exclusive to patients progressing to increasingly severe HIV-1-associated immunologic and neurologic disease. PMID- 12587071 TI - Multiple sclerosis-associated retrovirus particles cause T lymphocyte-dependent death with brain hemorrhage in humanized SCID mice model. AB - A retroviral element (multiple sclerosis-associated retrovirus, MSRV) defining a family of genetically inherited endogenous retroviruses (human endogenous retrovirus type W, HERV-W) has been characterized in cell cultures from patients with multiple sclerosis. Recently, MSRV retroviral particles or the envelope recombinant protein were shown to display superantigen activity in vitro, but no animal model has yet been set up for studying the pathogenicity of this retrovirus. In the present study, the pathogenicity of different sources of MSRV retroviral particles has been evaluated in a hybrid animal model: severe combined immunodeficiency (SCID) mice grafted with human lymphocytes and injected intraperitoneally with MSRV virion or mock controls. MSRV-injected mice presented with acute neurological symptoms and died within 5 to 10 days post injection. Necropsy revealed disseminated and major brain hemorrhages, whereas control animals did not show abnormalities (P <.001). In ill animals, reverse transcriptase-polymerase chain reaction (RT-PCR) analyses showed circulating MSRV RNA in serum, whereas overexpression of proinflammatory cytokines such as tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma was evidenced in spleen RNA. Neuropathological examination confirmed that hemorrhages occurred prior to death in multifocal areas of brain parenchyma and meninges. Further series addressed the question of immune-mediated pathogenicity, by inoculating virion to SCID mice grafted with total and T lymphocyte-depleted cells in parallel: dramatic and statistically significant reduction in the number of affected mice was observed in T-depleted series (P <.001). This in vivo study suggests that MSRV retroviral particles from MS cultures have potent immunopathogenic properties mediated by T cells compatible with the previously reported superantigen activity in vitro, which appear to be mediated by an overexpression of proinflammatory cytokines. PMID- 12587072 TI - Elevated peripheral benzodiazepine receptor expression in simian immunodeficiency virus encephalitis. AB - Measurement of central nervous system (CNS) expression of the peripheral benzodiazepine receptor (PBR), a microglia and macrophage activation marker, by positron emission tomography (PET) would aid clinical management of human immunodeficiency virus (HIV)-infected patients. To evaluate the utility of examining PBR expression in the CNS as a cellular activation marker in HIV CNS disease, PBR levels were measured in frontal cortex of simian immunodeficiency virus (SIV)-infected macaques with encephalitis and uninfected animals via PK11195 ligand autoradiography. [(3)H]-(R)-PK11195 binding to both grey matter (P =.017) and white matter (P =.038) was significantly higher in animals with SIV encephalitis (n = 10) versus control animals (n = 3). When PK11195 binding was compared with other microglial/macrophage activation markers (obtained via quantitative image analysis), a strong, significant association was found for both HAM56 (P =.004) and KP-1 (anti-CD68; P =.006) immunostaining in white matter. In contrast, grey matter PK11195 binding did not correlate with HAM56 (P =.46), KP-1 (P =.06), or glial fibrillary acidic protein (GFAP) immunostaining for astrocytic activation (P =.09). The regional nature of these increases in activation within the brain illustrates the crucial need to focus functional neuroimaging analyses of HIV-infected individuals on subcortical white matter to assess activation of microglia and macrophages. PMID- 12587073 TI - Herpes simplex virus type 1-induced encephalitis has an apoptotic component associated with activation of c-Jun N-terminal kinase. AB - Herpes simplex virus type 1 (HSV-1) triggered apoptosis in hippocampal cultures, as determined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and immunohistochemistry with antibody specific for the large fragment of activated caspase 3. The levels of phosphorylated (activated) c-Jun N terminal kinase (JNK) were also increased in HSV-1-infected hippocampal cultures as were the levels of activated c-Jun, its target. JNK activation was involved in HSV-1-induced apoptosis as evidenced by apoptosis inhibition with the JNK inhibitor SP600125. HSV-2 activated the mitogen-activated protein kinase/extracellular regulated protein kinase (MEK/ERK) survival pathway and did not trigger apoptosis in hippocampal cultures. The MEK specific inhibitor U0126 inhibited ERK activation and caused a significant increase in the percent TUNEL(+) cells in HSV-2-infected cultures, indicating that the failure of HSV-2 to trigger apoptosis is due to its ability to activate the MEK/ERK survival pathway. JNK was also activated in brain tissues from patients with HSV associated acute focal encephalitis (HSE) that were positive for HSV-1 antigen. JNK activation correlated with apoptosis, as determined by immunohistochemistry with antibody to activated caspase 3 or cleaved poly (ADP-ribose) polymerase (PARP). The data suggest that HSE has an apoptotic component that may contribute to disease pathogenesis. PMID- 12587074 TI - Multiple sclerosis-associated virus-related pol sequences found both in multiple sclerosis and healthy donors are more frequently expressed in multiple sclerosis patients. AB - In the etiopathogenesis of multiple sclerosis (MS), both genetic and environmental factors play an important role. Among environmental factors, viral infections are most likely connected with the etiology of MS. There are many evidence suggesting possible involvement of retroviruses in the development of autoimmune diseases including MS. Multiple sclerosis-associated retrovirus (MSRV) seems to be the important candidate for viral etiology of MS. The aim of the study was to analyze MSRV pol sequences in patients with MS. As control, groups of myasthenia gravis, Parkinson's disease, and migraine patients, and healthy individuals have been studied. The MSRV pol sequences have been analyzed in RNA isolated from the serum and in DNA and RNA of peripheral blood lymphocytes from untreated MS patients and control groups. The MSRV pol sequences have been detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and PCR technique, using specific oligonucleotide primers. In the serum RNA (cDNA), MSRV pol sequences have been identified in 31/32 MS patients. MSRV pol sequences were detected in serum cDNA of 9/17 myasthenia gravis patients, 7/16 Parkinson's disease patients, 10/21 migraine patients, and 13/27 healthy individuals. MSRV pol sequences were observed also in RNA from lymphocytes of all MS patients, 12/17 myasthenia gravis patients, 9/16 Parkinson's disease patients, 14/21 migraine patients, and 18/27 healthy donors. In the DNA from peripheral blood lymphocytes of all studied patients and healthy individuals, MSRV pol sequences have been found. The observed pattern of fiber-fluorescence in situ hybridization (FISH) signals suggests the presence of multiple copies of MSRV pol sequences, most likely tandemly dispersed in the genome. It can be concluded that MSRV pol sequences are endogenous, widespread in lymphocytes DNA, and transcribed into RNA of MS patients as well as of other studied patients and healthy individuals. However, more frequent expression of MSRV sequences detected in lymphocytes RNA (cDNA), as well as their presence in higher frequency in the serum of MS patients, may suggest the involvement of MSRV in the etiopathogenesis on MS. PMID- 12587076 TI - Molecular characterization of an Enterovirus 71 causing neurological disease in Germany. AB - Enterovirus 71 (EV71) is mainly known as a cause of hand-foot-and-mouth disease (HFMD) but sometimes associated with neurological disease, even as fatal brainstem encephalitis. In Europe, EV71 infections are extremely rare, in contrast to the worldwide situation. This is the first report of molecular characterization of an EV71 strain isolated in Europe that had caused neurological disease. The german strain is closest related to sublineage B2 strains isolated in the United States, which where mainly associated with neurological disease. Phylogenetic analysis also showed that the strain must have been imported to Germany several years ago, and continues to circulate since then. PMID- 12587075 TI - Experimental herpes simplex virus encephalitis: a combination therapy of acyclovir and glucocorticoids reduces long-term magnetic resonance imaging abnormalities. AB - Despite early antiviral treatment, herpes simples virus encephalitis (HSVE) still remains a life-threatening sporadic disease with high mortality and morbidity. In patients and in experimental disease, chronic progressive magnetic resonance imaging (MRI) abnormalities have been found even after antiviral therapy. Secondary autoimmune-mediated and not directly virus-mediated mechanisms might play a key role for the outcome of disease. This study aimed to evaluate a possible beneficial effect of a therapy of acyclovir and corticosteroids versus acyclovir only. In a mouse model of HSVE (intranasal inoculation with 10(5) pfu [plaque-forming units] of HSV-1 strain F), a long-term MRI study was realized. Cranial MRI was performed serially at days 2, 7, 14, 21, 60, and 180 in different therapy groups: 1, saline; 2, acyclovir; 3, acyclovir, subsequently methylprednisolone; 4, sham-infected with saline. Brain viral load peaked at day 7 to decline thereafter to a low baseline value. Viral load in group 1 was significantly higher than in animals with antiviral therapy. In group 4, no viral DNA was detectable. Viral load did not differ significantly between acyclovir and acyclovir/corticosteroid-treated groups, suggesting that the use of corticosteroids in addition to acyclovir does not increase viral burden. MRI findings in untreated and acyclovir-treated animals revealed chronic progressive changes. In contrast, there was a significant reduction of the severity of long term MRI abnormalities in acyclovir/corticosteroid-treated animals. With respect to abnormal MRI findings, this study demonstrates a clear beneficial effect of an acyclovir and corticosteroid therapy without influencing brain viral load. PMID- 12587077 TI - Neurological complications of varicella-zoster virus in human immunodeficiency virus-infected patients: changes in prevalence and diagnostic utility of polymerase chain reaction in cerebrospinal fluid. AB - Neurological complications caused by varicella-zoster virus (VZV) were diagnosed in 13 human immunodeficiency virus (HIV)-infected patients in our hospital. There was a favorable influence of highly active antiretroviral therapy (HAART) in the prevalence of these disorders among acquired immunodeficiency syndrome (AIDS) patients: Since 1996, only 1 of 961 AIDS patients had VZV neurological disease, compared to 9 of 1088 patients before that year (P =.02). Polymerase chain reaction (PCR) detected VZV DNA in cerebrospinal from 4 of 5 patients with VZV neurological disease, and from 2 of 130 HIV-infected patients with other neurological diseases (sensitivity 0.8, specificity 0.98 [95% confidence intervals 0.45-1 and 0.96-1, respectively], positive predictive value 0.94). PMID- 12587078 TI - Search for human herpesvirus 6 DNA in cerebrospinal fluid from AIDS patients with progressive multifocal leukoencephalopathy. PMID- 12587080 TI - Bypass balloon pumping: a newly developed device for mechanical circulatory support in heart failure. AB - Various types of assist devices have been developed for severe heart failures. Among them, intra-aortic balloon pumping (IABP) has achieved popularity, mainly in the treatment of patients with either severe cardiac infarction or low cardiac output syndrome (LOS) after open-heart surgery. However, IABP has a limitation in that although it acts as a pressure support, it cannot directly support flow volume. In cases of severe pump failure, stronger pump support would be required. However, there is a limit in the clinical use of temporary mechanical support using a pump because of the costs of the expensive equipment. With this in mind, we came up with a new idea in which the motion of a balloon in IABP system is converted into a power source for creating a one-way stream. In order to realize our idea, we made a J-shaped sample model of pulsating chamber incorporating two ball valves. In the hydrodynamic experiment using our new device, flows over 2500 ml/min (Max. 3475 ml/min) were obtained. The percent changes in parameters such as TTI, DPTI and bypass flows obtained by the experiment of left ventricular bypass have demonstrated that our assist system is effective for reducing cardiac work. PMID- 12587081 TI - Thoracic epidural anesthesia does not influence the incidence of postoperative atrial fibrillation after beating heart surgery. AB - BACKGROUND: At least 20 - 30 % of patients undergoing coronary artery bypass graft surgery (CABG) or beating-heart surgery develop postoperative atrial fibrillation (AF). We evaluated the effect of thoracic epidural anesthesia (TEA) on the occurrence of postoperative AF in patients submitted to CABG without cardiopulmonary bypass (OPCABG). METHODS: We performed a retrospective analysis of 125 patients undergoing myocardial revascularization. Early postoperative incidence of AF was compared between three groups of patients - 50 after conventional CABG, 45 after OPCABG, and 30 after OPCABG combined with TEA intraoperatively and postoperatively. Clinical profile of the patients, including factors with a potential influence on postoperative AF was matched for groups. RESULTS: Postoperative AF occurred in 13.3 % of the TEA-treated patients, in 17.7 % of the patients in the OPCABG group, and in 26 % of the patients in the CABG group. This difference did not carry any statistical significance. Risk factors and incidence of postoperative complications were comparable in all groups. CONCLUSION: TEA has no effect on the incidence of postoperative AF in patients undergoing beating-heart surgery. PMID- 12587082 TI - Impact of diabetes mellitus on cardiac surgery outcome. AB - BACKGROUND: Diabetes mellitus is an established independent risk factor related to significant morbidity and mortality after cardiac surgical procedures. METHODS: Data on 16,184 patients undergoing cardiac surgery with and without cardiopulmonary bypass between April 1996 and August 2001 were prospectively evaluated. Diabetes mellitus as a patient related risk factor was subjected to univariate analysis to identify potential associations to 28 intra- and postoperative outcome variables. Outcome variables having a significant association with diabetes mellitus (p < 0.05) were then subjected to a stepwise logistic regression model to identify the influence of diabetes mellitus as compared to additional 30 different patient related risk factors and treatment variables. Diabetes mellitus was defined as glucose intolerance treated either dietary, with oral hypoglycemics or with insulin. RESULTS: Overall prevalence of diabetes mellitus was 33.3 %. Compared to non-diabetic patients the group with diabetes mellitus was older (p < 0.0001) and had a significantly lower ejection fraction (p < 0.0001). 15 outcome variables having a significant association with diabetes mellitus were identified. Furthermore, diabetes mellitus could be identified as an independent predictor for 7 postoperative outcome variables (prolonged ICU-stay, sternal instability and/or infection, sternal revision and refixation respiratory insufficiency, postoperative delirium, perioperative stroke, renal dysfunction, postoperative reintubation). CONCLUSION: Diabetes mellitus is a significant independent predictor for several postoperative outcome variables after cardiac surgery associated with higher postoperative morbidity and prolonged hospital stay. PMID- 12587083 TI - Predictors of homologous blood transfusion for patients undergoing open heart surgery. AB - OBJECTIVE: It has become very important to avoid homologous blood transfusions in today's cardiac surgery. We performed a retrospective analysis to find out preoperative factors to predict the risk for transfusion of red-cell concentrate in cardiac surgery. METHODS: This study included 400 consecutive patients undergoing coronary artery bypass grafting. We also included emergency (4 %) and re-operations (8 %). We tried to find out predictive factors for the need of transfusion of red-cell concentrate on the base of logistic regression coefficient and the odds ratio. We looked at the following factors as predictors of transfusion risk: left ventricular ejection fraction < 0.35, age over 70 years, preoperative hemoglobin < 11 g/dl, insulin-dependent diabetes (IDDM), emergency operation, female sex, impaired renal function (creatinine > 1.6 mg/dl), and re-operation. RESULTS: In our group, 132 (33 %) patients received transfusion during hospitalization, while 268 (67 %) did not. On average, 2.2 +/- 0.68 units of red-cell concentrate were transfused per patient. In addition, we found a predictive value for transfusion for the following parameters: age > 70 years, preoperative hemoglobin < 11 g/dl, re-operation and ejection fraction < 0.35. We could not find any significantly increased blood transfusion risk in female cases, insulin dependent diabetes mellitus, or impaired renal function. CONCLUSIONS: We could show that there is normally no need for blood transfusion in (2/3) of the patients in cardiac surgery according to this study's results. Furthermore, it was obvious that some patient variables can be used predict the risk for perioperative transfusion. Based on these results, the prophylactic administration of aprotinin or the use of a cell saver could be useful in selected patients. PMID- 12587084 TI - Simultaneous carotid endarterectomy and cardiac surgery--additional risk factor or safety procedure? AB - BACKGROUND: The occurrence of severe carotid artery disease in more than 12 % of patients requiring CABG results in a discrepancy concerning the best treatment for both diseases. We reviewed the early outcome of patients with CABG and/or valve replacement and simultaneous carotid endarterectomy (CEA). METHODS: We evaluated retrospectively 244 patients operated simultaneously between 7/94 and 10/2001. 209 patients received CABG; 35 patients CABG and/or valve replacement. Mean age was 68 years. 188 patients were male. We analyzed risk factors, morbidity, incidence of neurological complications and 30 day mortality. RESULTS: Perioperative stroke with hemiplegia occurred in 3.3 % (8 patients). 4 of these patients showed contralateral carotid artery occlusion, 2 contralateral severe stenosis. 2 patients (0.8 %) experienced prolonged reversible ischemic neurological deficit (PRIND), 4 patients (1.6 %) transient ischemic attack (TIA). 30-day lethality was 4.5 %. 3 patients died due to low cardiac output, 6 patients due to extracardial reasons, 2 patients (0.8 %) developed a cerebral death. CONCLUSIONS: Simultaneous CEA and cardiac surgery can be performed with an acceptable risk for neurological complications and mortality. Occlusion of contralateral carotid artery could be identified as an evident predictor for increased neurological complications. Compared to two-stage procedures, combined operations yield a reduction of hospital costs. PMID- 12587085 TI - Multiple minimally invasive direct CABG for the complete revascularization: the figure L approach. AB - BACKGROUND: Coronary artery bypass grafting on the beating heart via median sternotomy causes significant hemodynamic compromise during displacement of the heart, especially in patients with low ejection fraction or those who have potential catastrophic sternal reentry injuries. We have developed an innovative minimally invasive surgical approach (figure L Approach) for the treatment of multi-vessel coronary artery disease; especially in cases with huge hearts (> CTR 50 %) including tracheostomy requiring COLD, and redo cases. The objective of this study was to assess the efficacy and safety of this alternative surgical incision. METHODS: From January 1998 to March 2001, 22 patients underwent complete revascularization using this figure L Approach. Left submammary anterior thoracotomy incisions are made with the medial part of the incision vertically extending downward to the upper abdomen. The costal margin was divided. The pleural and peritoneal spaces were then entered. A chest retractor was placed to elevate the chest wall, exposing the heart and stomach. Two to four arterial grafts including LIT A, RGEA, RA and IEA were harvested. Revascularization of the LAD, LCX and distal RCA were performed in 22 patients without cardiopulmonary bypass on complex performed arterial grafts using this approach. In four patients, triple and quadruple vessel grafting was performed. RESULTS: No early deaths or postoperative complications occurred. The mean coronary clamp time was 12.5 +/- 5.1 minutes for the LAD, 10.8 +/- 1.8 minutes for diagonal branch, 14.1 +/- 2.9 minutes for the LCX and 16.0 +/- 5.1 minutes RCA time. There were no late deaths or angina during the mean follow-up of 37.3 +/- 9.0 months. Postoperative coronary angiography demonstrated widely patent grafts in all but one patient. Hemodynamics did not change significantly during the distal anastomoses. No wound healing problems were experienced. CONCLUSIONS: Multiple MIDCABG using the figure L Approach represents a novel way of approaching both the LAD, CX and distal RCA in patients with multi-vessel disease, especially in those with low ejection fraction, huge heart (> CTR 50 %), or hemodynamically unstable patients, without any hemodynamic deterioration or sternal related complication. PMID- 12587086 TI - Predictors of perioperative mortality after coronary artery bypass grafting in the elderly. AB - BACKGROUND: Coronary artery bypass grafting (CABG) is associated with higher operative risk in the elderly compared to younger patients. The aim of this study was to evaluate risk factors for perioperative mortality after CABG in the elderly. METHODS: We investigated 325 consecutive patients aged 75 or over undergoing isolated CABG at our institution. We analyzed the patients' characteristics and perioperative outcome. Patients were divided into survivors and non-survivors; risk factors and complications were compared. Based on this, we performed a multivariate logistic regression analysis to determine independent risk factors for perioperative mortality. RESULTS: Non-survivors of CABG more often suffered from concomitant extracardiac atherosclerosis (non-survivors, 62.2 %; survivors, 40.6 %; p = 0.013) as well as from renal insufficiency preoperatively (non-survivors, 35.1 %; survivors 8.0 %; p < 0.0001). A trend towards higher incidences of impaired left ventricular function (non-survivors, 37.8 %; survivors, 29.9 %; p = 0.105) and a history of recent myocardial infarction (non-survivors, 29.7 %; survivors, 17.0 %; p = 0.061) were found in non-survivors compared to survivors. Furthermore, non-survivors more often underwent CABG with cardiopulmonary bypass (CPB non-survivors, 96.1 %; survivors 70.6 %; p = 0.0005). Multivariate logistic regression analysis revealed that preoperatively impaired renal function (OR: 2.857, p < 0.0001), use of CPB (OR: 5.952, p = 0.0175), extracardiac atherosclerosis (OR: 1.581, p = 0.0228), and recent myocardial infarction (OR: 1.574, p = 0.0405) were independent risk factors for perioperative mortality. Comparison of patients undergoing CABG with or without CPB reveals that patients operated without CPB had a higher preoperative risk than patients undergoing CABG with CPB. CONCLUSION: These results show that besides impaired renal function, extracardiac atherosclerosis, and history of recent myocardial infarction, the use of CPB is a major risk factor for CABG in the elderly. Perioperative mortality and morbidity can be significantly reduced if CPB is avoided. PMID- 12587087 TI - Pneumonectomy for lung metastases: report of ten cases. AB - Today, pulmonary resection for lung metastases is a widely accepted treatment if complete resection can be achieved. However, 5-year survival is only 40 %. Many patients develop recurrences, but some reports have demonstrated that salvage operations can result in a long-term survival. A resection of a complete lung or a resection of more than a lung is still controversial since procedure-related morbidity or mortality does not outweigh the survival benefit. We report on a series of 10 consecutive patients who underwent a primary pneumonectomy or an operation on the residual lung after pneumonectomy with curative intent for pulmonary metastases. 5 year survival rates for the 10 patients after pneumonectomy alone or with additional resection was 45 %, which was not significantly different from those who underwent a more minor resection with a 5 year survival of 39 % (p = 0.40). Since there is currently no alternative proven therapy for patients with isolated pulmonary metastases, a primary or completion pneumonectomy may be offered to selected patients as long as sufficient pulmonary reserve is present, and a complete resection can be achieved. PMID- 12587088 TI - Unusual complication of sternotomy: bone fragment induced right ventricular rupture after mitral valve replacement surgery. AB - Major surgical complications following open cardiac procedures via median sternotomy are infrequent but potentially devastating events. We report on a unique, fatal complication of median sternotomy. A 44-year-old woman underwent mitral valve replacement for endocarditis related to intravenous drug abuse. Twenty days after the surgery, she presented to the emergency department in acute distress, and died of cardiac tamponade soon after admission. Postmortem examination revealed a defect in the right ventricular wall caused by a bone fragment resulting from the median sternotomy. PMID- 12587089 TI - Morgagni hernia and thoracic deformities. AB - We report two cases of Morgagni hernia associated with pectus carinatum. This association is exceptional; only two other cases have been reported so far. In one of our patients, an abdominal surgical approach was used to repair the Morgagni hernia and to perform a Nissen-Rossetti procedure (for an associated endobrachyesophagus); the patient did not require correction of the pectus carinatum. In the other patient, both thoracic deformity and Morgagni hernia were repaired using the same thoracic approach. PMID- 12587090 TI - A simple method to reduce ischemic time of the spinal cord in extensive thoracoabdominal aneurysm operations. AB - OBJECTIVE: Although the incidence of paraplegia from surgery of extensive thoracoabdominal aneurysms Crawford type I - Ill.could be reduced to about 10 % in more recent series, development of additional measures to avoid this disastrous complication seems worthwhile. One of of the major determinants of preventing ischemia is obviously keeping ischemic time of the spinal cord below 30'. In this regard, we introduced a surgical technique that allowed reperfusing the intercostal arteries within 30 min. METHODS: In a consecutive series 17 Patients with thoracoabdominal aneurysms (9 Crawford type II, 7 type I and one type III) underwent thoracoabdominal aortic replacement using either fem.-fem.-or left atrio-fem. bypass with selective perfusion of the visceral vessels. To keep the ischemic time of the spinal cord as short as possible, the first step of aortic replacement consisted of reattachement of the intercostal arteries of the segments Th 10-L 1 followed by immediate reperfusion via a side branch of the prostheses with the aid of the heart and lung machine. RESULTS: Mean ischemic time was 25 min. The mean number of reperfused intercostal arteries was five. One patient died of bleeding complications and could not be evaluated for paraplegia. There was no further hospital mortality. One patient developed paraplegia. With the exception of two patients, all had a very fast recovery and limited stay on the ICU. CONCLUSIONS: Compared to our former experience, we were able to reduce the incidence of paraplegia dramatically. Our observations to date strongly support the hypothesis that an ischemic period of less than 30' is fairly well tolerated by the spinal cord. PMID- 12587091 TI - Novadaq Spy Intraoperative Imaging System--current status. AB - The Novadaq Spy Intraoperative Imaging System is able to depict a fluorescent contrast agent emitting light at 830 nm when passing through the vascular tree or through myocardial chambers. The passage of the contrast agent can be observed in real time, allowing quality control in adult and congenital cardiovascular surgery. PMID- 12587092 TI - Cerebral oxygen monitoring during neonatal cardiopulmonary bypass and deep hypothermic circulatory arrest. PMID- 12587093 TI - Accumulation of mitochondrial DNA mutations in human immunodeficiency virus infected patients treated with nucleoside-analogue reverse-transcriptase inhibitors. AB - Nucleoside reverse-transcriptase inhibitor (NRTI) therapy for human immunodeficiency virus (HIV) infection has been associated with mitochondrial DNA (mtDNA) polymerase-gamma inhibition and subsequent mtDNA depletion. Effects on mtDNA mutation, although suggested by critical involvement of polymerase-gamma in DNA-repair reactions, are unknown. In the present study, we assessed the nature and frequency of mitochondrial genome sequence differences in peripheral-blood samples taken prior to NRTI therapy and after 6-77 mo of treatment in 16 NRTI treated patients. Samples from 10 HIV-infected, treatment-naive control individuals were taken at similar time intervals. Single-stranded conformation polymorphism (SSCP) and DNA-sequencing analysis techniques were used to detect mitochondrial genome sequence variants between paired longitudinal samples, and heteroplasmic populations were quantified after cloning and repeat SSCP/sequencing. Of 16 individuals treated with NRTIs, 5 exhibited altered SSCP profiles associated with the development of novel heteroplasmic DNA sequence changes, whereas no SSCP pattern change within these regions was observed in the control individuals. Heteroplasmic sequence changes were distributed across four regions of the genome: the noncoding region to 12S ribosomal RNA, reduced nicotinamide-adenine-dinucleotide dehydrogenase 1, and cytochrome oxidase subunits I and III. Of the total of 26 patients who were examined in the present study, 4 of 5 patients with detectable mtDNA sequence changes since commencement of therapy developed evidence of peripheral fat wasting (lipoatrophy) between sample intervals (P=.031). One patient, without detectable sequence changes on NRTI therapy, also developed lipoatrophy. Levels of mtDNA copies/cell in blood samples were determined by quantitative PCR for 11 of the 16 NRTI-exposed patients; 7 of these 11 patients showed reduced levels of mtDNA in blood after therapy, including all 3 patients tested with evidence of mtDNA sequence changes on therapy. These data indicate that NRTI therapy provides conditions permissive for the development of peripheral-blood mtDNA mutations in vivo. PMID- 12587094 TI - Use of multivariate linkage analysis for dissection of a complex cognitive trait. AB - Replication of linkage results for complex traits has been exceedingly difficult, owing in part to the inability to measure the precise underlying phenotype, small sample sizes, genetic heterogeneity, and statistical methods employed in analysis. Often, in any particular study, multiple correlated traits have been collected, yet these have been analyzed independently or, at most, in bivariate analyses. Theoretical arguments suggest that full multivariate analysis of all available traits should offer more power to detect linkage; however, this has not yet been evaluated on a genomewide scale. Here, we conduct multivariate genomewide analyses of quantitative-trait loci that influence reading- and language-related measures in families affected with developmental dyslexia. The results of these analyses are substantially clearer than those of previous univariate analyses of the same data set, helping to resolve a number of key issues. These outcomes highlight the relevance of multivariate analysis for complex disorders for dissection of linkage results in correlated traits. The approach employed here may aid positional cloning of susceptibility genes in a wide spectrum of complex traits. PMID- 12587095 TI - A tobit variance-component method for linkage analysis of censored trait data. AB - Variance-component (VC) methods are flexible and powerful procedures for the mapping of genes that influence quantitative traits. However, traditional VC methods make the critical assumption that the quantitative-trait data within a family either follow or can be transformed to follow a multivariate normal distribution. Violation of the multivariate normality assumption can occur if trait data are censored at some threshold value. Trait censoring can arise in a variety of ways, including assay limitation or confounding due to medication. Valid linkage analyses of censored data require the development of a modified VC method that directly models the censoring event. Here, we present such a model, which we call the "tobit VC method." Using simulation studies, we compare and contrast the performance of the traditional and tobit VC methods for linkage analysis of censored trait data. For the simulation settings that we considered, our results suggest that (1) analyses of censored data by using the traditional VC method lead to severe bias in parameter estimates and a modest increase in false-positive linkage findings, (2) analyses with the tobit VC method lead to unbiased parameter estimates and type I error rates that reflect nominal levels, and (3) the tobit VC method has a modest increase in linkage power as compared with the traditional VC method. We also apply the tobit VC method to censored data from the Finland-United States Investigation of Non-Insulin-Dependent Diabetes Mellitus Genetics study and provide two examples in which the tobit VC method yields noticeably different results as compared with the traditional method. PMID- 12587097 TI - Undetected genotyping errors cause apparent overtransmission of common alleles in the transmission/disequilibrium test. AB - The transmission/disequilibrium test (TDT), a family-based test of linkage and association, is a popular and intuitive statistical test for studies of complex inheritance, as it is nonparametric and robust to population stratification. We carried out a literature search and located 79 significant TDT-derived associations between a microsatellite marker allele and a disease. Among these, there were 31 (39%) in which the most common allele was found to exhibit distorted transmission to affected offspring, implying that the allele may be associated with either susceptibility to or protection from a disease. In 27 of these 31 studies (87%), the most common allele appeared to be overtransmitted to affected offspring (a risk factor), and, in the remaining 4 studies, the most common allele appeared to be undertransmitted (a protective factor). In a second literature search, we identified 92 case-control studies in which a microsatellite marker allele was found to have significantly different frequencies in case and control groups. Of these, there were 37 instances (40%) in which the most common allele was involved. In 12 of these 37 studies (32%), the most common allele was enriched in cases relative to controls (a risk factor), and, in the remaining 25 studies, the most common allele was enriched in controls (a protective factor). Thus, the most common allele appears to be a risk factor when identified through the TDT, and it appears to be protective when identified through case-control analysis. To understand this phenomenon, we incorporated an error model into the calculation of the TDT statistic. We show that undetected genotyping error can cause apparent transmission distortion at markers with alleles of unequal frequency. We demonstrate that this distortion is in the direction of overtransmission for common alleles. Therefore, we conclude that undetected genotyping errors may be contributing to an inflated false positive rate among reported TDT-derived associations and that genotyping fidelity must be increased. PMID- 12587096 TI - Reciprocal and nonreciprocal recombination at the glucocerebrosidase gene region: implications for complexity in Gaucher disease. AB - Gaucher disease results from an autosomal recessive deficiency of the lysosomal enzyme glucocerebrosidase. The glucocerebrosidase gene is located in a gene-rich region of 1q21 that contains six genes and two pseudogenes within 75 kb. The presence of contiguous, highly homologous pseudogenes for both glucocerebrosidase and metaxin at the locus increases the likelihood of DNA rearrangements in this region. These recombinations can complicate genotyping in patients with Gaucher disease and contribute to the difficulty in interpreting genotype-phenotype correlations in this disorder. In the present study, DNA samples from 240 patients with Gaucher disease were examined using several complementary approaches to identify and characterize recombinant alleles, including direct sequencing, long-template polymerase chain reaction, polymorphic microsatellite repeats, and Southern blots. Among the 480 alleles studied, 59 recombinant alleles were identified, including 34 gene conversions, 18 fusions, and 7 downstream duplications. Twenty-two percent of the patients evaluated had at least one recombinant allele. Twenty-six recombinant alleles were found among 310 alleles from patients with type 1 disease, 18 among 74 alleles from patients with type 2 disease, and 15 among 96 alleles from patients with type 3 disease. Several patients carried two recombinations or mutations on the same allele. Generally, alleles resulting from nonreciprocal recombination (gene conversion) could be distinguished from those arising by reciprocal recombination (crossover and exchange), and the length of the converted sequence was determined. Homozygosity for a recombinant allele was associated with early lethality. Ten different sites of crossover and a shared pentamer motif sequence (CACCA) that could be a hotspot for recombination were identified. These findings contribute to a better understanding of genotype-phenotype relationships in Gaucher disease and may provide insights into the mechanisms of DNA rearrangement in other disorders. PMID- 12587098 TI - Controlling type I error rate for fast track drug development programmes. AB - The U.S. Food and Drug Administration (FDA) Modernization Act of 1997 has a Section (No. 112) entitled 'Expediting Study and Approval of Fast Track Drugs' (the Act). In 1998, the FDA issued a 'Guidance for Industry: the Fast Track Drug Development Programs' (the FTDD programmes) to meet the requirement of the Act. The purpose of FTDD programmes is to 'facilitate the development and expedite the review of new drugs that are intended to treat serious or life-threatening conditions and that demonstrate the potential to address unmet medical needs'. Since then many health products have reached patients who suffered from AIDS, cancer, osteoporosis, and many other diseases, sooner by utilizing the Fast Track Act and the FTDD programmes. In the meantime several scientific issues have also surfaced when following the FTDD programmes. In this paper we will discuss the concept of two kinds of type I errors, namely, the 'conditional approval' and the 'final approval' type I errors, and propose statistical methods for controlling them in a new drug submission process. PMID- 12587099 TI - Stopping clinical trials because of treatment ineffectiveness: a comparison of a futility design with a method of stochastic curtailment. AB - This paper introduces a simple futility design that allows a comparative clinical trial to be stopped due to lack of effect at any of a series of planned interim analyses. Stopping due to apparent benefit is not permitted. The design is for use when any positive claim should be based on the maximum sample size, for example to allow subgroup analyses or the evaluation of safety or secondary efficacy responses. A final frequentist analysis can be performed that is valid for the type of design employed. Here the design is described and its properties are presented. Its advantages and disadvantages relative to the use of stochastic curtailment are discussed. PMID- 12587100 TI - Sequential designs for phase III clinical trials incorporating treatment selection. AB - Most statistical methodology for phase III clinical trials focuses on the comparison of a single experimental treatment with a control. An increasing desire to reduce the time before regulatory approval of a new drug is sought has led to development of two-stage or sequential designs for trials that combine the definitive analysis associated with phase III with the treatment selection element of a phase II study. In this paper we consider a trial in which the most promising of a number of experimental treatments is selected at the first interim analysis. This considerably reduces the computational load associated with the construction of stopping boundaries compared to the approach proposed by Follman, Proschan and Geller (Biometrics 1994; 50: 325-336). The computational requirement does not exceed that for the sequential comparison of a single experimental treatment with a control. Existing methods are extended in two ways. First, the use of the efficient score as a test statistic makes the analysis of binary, normal or failure-time data, as well as adjustment for covariates or stratification straightforward. Second, the question of trial power is also considered, enabling the determination of sample size required to give specified power. PMID- 12587101 TI - Comparison of stratification and adaptive methods for treatment allocation in an acute stroke clinical trial. AB - Achieving balance on prognostic factors between treatment groups in a clinical trial is important to ensure that any observed treatment effect may be attributed to the treatment itself. Improving the balance on prognostic factors also potentially increases the statistical power attained in a trial. Substantial imbalances may occur by chance if simple randomization is used. Allocation of the treatment according to stratified random blocks based on clinical features is the conventional approach to obtain treatment groups that are as similar as possible. An alternative approach, known as minimization (or more generally as adaptive stratification), has also been proposed. We assessed the feasibility of adaptive stratification in the context of a clinical trial of insulin to control plasma glucose level following acute stroke. We determined suitable settings for the parameters in the adaptive stratification procedure by simulation studies. Specifically, we assessed: the optimal probability for allocating a patient to the preferred (leading to least imbalance on prognostic factors) treatment group; the number of variables that could be incorporated in the adaptive stratification algorithm; the weighting that should be given to each variable; and whether interactions between variables should be included. We then compared the statistical power, across a range of simulated treatment effects, between trials where treatments were allocated by stratified random blocks and by adaptive stratification. Finally, we considered the importance of the method of analysis in realizing the gain in power which may potentially be achieved by allocating treatments using stratified random blocks or adaptive stratification. PMID- 12587102 TI - Group sequential design for comparative diagnostic accuracy studies. AB - In the field of diagnostic medicine, comparative clinical trials are necessary for assessing the utility of one diagnostic test over another. The area under the receiver operating characteristic (ROC) curve, commonly referred to as AUC, is a general measure of a test's inherent ability to distinguish between patients with and without a condition. Standardized AUC difference is the most frequently used statistic for comparing two diagnostic tests. In therapeutic comparative clinical trials with sequential patient entry, fixed sample design (FSD) is unjustified on ethical and economical grounds and group sequential design (GSD) is frequently used. In this paper, we argue that the same reasoning exists for the comparative clinical trials in diagnostic medicine and hence GSD should be utilized in this field for designing trials. Since computation of the stopping boundaries of GSD and data analysis after a group sequential test rely heavily on Brownian motion approximation, we derive the asymptotic distribution of the standardized AUC difference statistic and point out its resemblance to the Brownian motion. Boundary determination and sample size calculation are then illustrated through an example from a cancer clinical trial. PMID- 12587103 TI - Therapeutic equivalence: fallacies and falsification. AB - The number of studies designed specifically to demonstrate therapeutic equivalence or alternatively non-inferiority of pharmaceutical treatments has increased dramatically in recent years, during which time awareness of the methodological issues has increased. Regulatory authorities have been quick to recognize the need for specific support and have either published or initiated the creation of relevant guidance. Common misconceptions prevail however regarding sample size estimation and the choice of the most appropriate patient population to analyse while other areas such as equivalence margin specification and covariate adjustment have been neglected. This paper challenges some of the regulatory advice and the interpretation that others have made of this guidance with the aim of stimulating further debate. PMID- 12587104 TI - Hierarchical Bayesian approaches to phase II trials in diseases with multiple subtypes. AB - We propose a methodology for conducting phase II clinical trials in settings where the disease is categorized into multiple subtypes. A hierarchical Bayesian model is assumed for treatment effects within the subtypes. The hierarchical model, which is tailored to each particular application, allows treatment effects to differ across subtypes while assuming a priori that the effects are exchangeable and correlated. Two applications are described. The first is a trial of imatinib for sarcoma in which treatment activity is characterized by a binary indicator of tumour response. The second is a phase II trial of a new preparative regimen for allogeneic bone marrow transplantation in patients with haematologic malignancies, with treatment effect characterized by the mean time from transplant to disease progression or death. The applications illustrate how the hierarchical Bayesian model borrows strength across subtypes. PMID- 12587105 TI - Adjusting treatment comparisons to account for non-randomized interventions: an example from an angina trial. AB - In a clinical trial where some subjects receive one or more non-randomized interventions during follow-up, primary interest is in the effect of the overall treatment strategies as implemented, but it may also be of interest to adjust treatment comparisons for non-randomized interventions. We consider non randomized interventions, especially surgical procedures, which only occur when the outcome would otherwise have been poor. Focusing on an outcome measured repeatedly over time, we describe the variety of questions that may be addressed by an adjusted analysis. The adjusted analyses involve new outcome variables defined in terms of the observed outcomes and the history of non-randomized intervention. We also show how to check the assumption that the outcome would otherwise have been poor, and how to do a sensitivity analysis. We apply these methods to a clinical trial comparing initial angioplasty with medical management in patients with angina. We find that the initial benefit of a single angioplasty in reducing angina tends to disappear with time, but a policy of additional interventions as required yields a benefit that is maintained over 4 years. Such methods may be of interest to many pragmatic randomized trials in which the effects of the initial randomized treatments and the effects of the overall treatment strategies as implemented are both of interest. PMID- 12587106 TI - Pattern mixture models for longitudinal quality of life studies in advanced stage disease. AB - Analyses of longitudinal quality of life (QOL) for patients with advanced stage disease are frequently plagued by problems of non-random drop-out attributable to deteriorating health and/or death. As an example, Moinpour et al. cite specific challenges which limited their report and assessment of QOL for patients treated for advanced stage colorectal cancer in a clinical trial of several chemotherapeutic regimes performed by the Southwest Oncology Group. A particular source of confusion that arises in studies of advanced stage disease is whether or not to differentiate loss of follow-up due to death from drop-out where the patient is still alive but has dropped from the study. In this paper we examine exploratory data techniques for longitudinal QOL data with non-random missingness due to drop-out and censorship by death. We propose a pattern mixture model for longitudinal QOL, time of drop-out and survival, which allows for straightforward implementation of sensitivity analyses and explicit comparisons to the raw data. Our method is illustrated in the context of analysing the data and addressing the challenges posed by Moinpour et al. PMID- 12587107 TI - Sensitivity analysis for the estimation of rates of change with non-ignorable drop-out: an application to a randomized clinical trial of the vitamin D3. AB - The vitamin D(3) trial was a repeated measures randomized clinical trial for secondary hyperparathyroidism in haemodialysis patients where the efficacy of the vitamin D(3) infusions for suppressing the secretion of parathyroid hormone (PTH) was compared among four dose groups over 12 weeks. In this trial, patients terminated the study before the scheduled end of the study due to their elevated serum calcium (Ca) level, that is, the administration of the vitamin D(3) was expected to cause hypercalcaemia as an adverse event. In this setting of monotone missingness, there is a potential for bias in estimation of mean rates of decline in PTH for each treatment group using the standard methods such as the generalized estimating equations (GEE) which ignore the observed past Ca histories. We estimated the treatment-group-specific mean rates of decline in PTH by the inverse probability of censoring weighted (IPCW) methods which account for the observed past histories of time-dependent factors that are both a predictor of drop-out and are correlated with the outcomes. The IPCW estimator can be viewed as an extension of the GEE estimator that allows for the data to be MAR but not MCAR. With missing data, it is rarely appropriate to analyse the data solely under the assumption that the missing data process is ignorable, because the assumption of ignorable missingness cannot be guaranteed to hold and is untestable from the observed data. We proposed a sensitivity analysis that examines how inference about the IPCW estimates of the treatment-group-specific mean rates of decline in PTH changes as we vary the non-ignorable selection bias parameter over a range of plausible values. PMID- 12587108 TI - Solid-state nuclear magnetic resonance spectroscopy--pharmaceutical applications. AB - Solid-state nuclear magnetic resonance (NMR) spectroscopy has become an integral technique in the field of pharmaceutical sciences. This review focuses on the use of solid-state NMR techniques for the characterization of pharmaceutical solids (drug substance and dosage form). These techniques include methods for (1) studying structure and conformation, (2) analyzing molecular motions (relaxation and exchange spectroscopy), (3) assigning resonances (spectral editing and two dimensional correlation spectroscopy), and (4) measuring internuclear distances. PMID- 12587109 TI - Establishment of a quantitative structure-activity relationship model for evaluating and predicting the protective potentials of phenolic antioxidants on lipid peroxidation. AB - Antioxidant activities of phenolic compounds have been extensively explored, but the determinant factors underlying their mechanisms of action remain to be elucidated. In the present work, a series of phenolic compounds (hydroxylated connamic, benzoic acid, and polyphenol) were studied for their protection against lipid peroxidation (LPO) in two model experiments, pre-emulsified linoleic acid system and phosphate buffered linolenic acid system. The mechanisms of action as well as activity determinants were investigated by computational chemistry and multiple-linear regression analysis. Upon elucidating the LPO inhibition properties and the relationship between their structural natures and antioxidant activities (SAR), a fairly satisfactory multidescriptor quantitative SAR model was derived, which extended our understanding of LPO inhibition mechanisms and should be valuable in assessing or predicting the anti-LPO activity of phenolic antioxidants. PMID- 12587110 TI - Impurities in a morphine sulfate drug product identified as 5-(hydroxymethyl)-2 furfural, 10-hydroxymorphine and 10-oxomorphine. AB - Stability testing of morphine sulfate formulated with nonpareil sugar seeds (consisting of sucrose and starch) and fumaric acid revealed the formation of the three impurities 5-(hydroxymethyl)-2-furfural, 10-hydroxymorphine and 10 oxomorphine. 5-(Hydroxymethyl)-2-furfural was isolated via semipreparative HPLC utilizing volatile mobile phase constituents and was identified by analysis of its HRMS and NMR spectra. 10-Hydroxymorphine and 10-oxomorphine were obtained via semipreparative HPLC and subsequent removal of ion-pair reagents using an anion exchange resin, or by solid phase extraction, and identified by spectroscopic analysis followed by comparison with authentic materials. 5-(Hydroxymethyl)-2 furfural is a degradation product of hexose sugars, and its formation in the presence of morphine sulfate formulated with fumaric acid suggests that caution should be exercised when including nonpareil seeds in formulations that contain acidic drug salts and/or acid excipients. The preliminary results of tests on the interaction of acidic salts of some other drugs with nonpareil seeds are presented. PMID- 12587111 TI - Characteristics of hydroxypropyl methylcellulose influencing compactibility and prediction of particle and tablet properties by infrared spectroscopy. AB - Particle characteristics, chemical substitution, compaction behavior, and tablet properties of hydroxypropyl methylcellulose powders from two different suppliers were related using multivariate data analysis. By Principal Component Analysis it was shown that the the degree of substitution of the HPMC powders did not correlate to the particle and compaction properties as strongly as anticipated. Particle shape and powder surface area seem to be more important for the compaction behaviour of the powders than the degree of substitution. In addition, particle and tablet properties were predicted from infrared spectral data. Fourier transform infrared (FTIR) and near infrared (NIR) spectral data of the powders were combined with measured values of the particle characteristics, compaction behavior, and tablet properties using the multivariate data analysis program SIMCA 7.1. Properties like density, particle shape, tablet tensile strength, and drug release characteristics of the HPMC powders and corresponding tablets in this study could be predicted using Partial Least Squares models. In conclusion, the particle shape and powder surface area of HPMC powders seem to be important factors for the quality of tablet attained. Further, this study confirms that NIR and FTIR analysis used in combination with multivariate analysis are powerful tools for predicting the properties of materials and the quality of the end product. PMID- 12587112 TI - Properties of rapidly dissolving eutectic mixtures of poly(ethylene glycol) and fenofibrate: the eutectic microstructure. AB - Poly(ethylene glycol) or PEG is an ideal inactive component for preparing simple binary eutectic mixtures because of its low entropy of fusion ( approximately 0.0076 J/mol-K), lower melting point (approximately 62 degrees C) compared to most pharmaceuticals, miscibility with drugs at elevated temperatures, and its covalent crystalline lattice. Implication of these physicochemical properties on eutectic crystallization and size reduction of the drug is discussed. Enhancement of the dissolution rate of a poorly soluble compound through the formation of PEG drug eutectics was investigated using fenofibrate. Solid dispersions of PEG fenofibrate when characterized, revealed that PEG and fenofibrate form a simple eutectic mixture containing 20-25%(w/w) fenofibrate at the eutectic point. Eutectic crystallization led to the formation of an irregular microstructure in which fenofibrate crystals were found to be less than 10 microm in size. Dissolution rate improvement of fenofibrate correlated with the phase diagram, and the amount of fenofibrate released from the dispersions that contained fenofibrate as a eutectic mixture was at least 10-fold higher compared to untreated fenofibrate. On aging, the dissolution rate of the dispersion containing 15%(w/w) fenofibrate in PEG remained unaltered. The results indicate that PEG-drug eutectic formation is a valuable option for particle size reduction and subsequent dissolution rate improvement. PMID- 12587113 TI - Effects of excipients on hydrate formation in wet masses containing theophylline. AB - Transformations between solid phases in dosage forms can lead to instability in drug release. Thus, it is important to understand mechanisms and kinetics of phase transformations and factors that may influence them. During wet granulation theophylline shows pseudopolymorphic changes that may alter its dissolution rate. The aim of this study was to investigate whether excipients, such as alpha lactose monohydrate or the highly water absorbing silicified microcrystalline cellulose (SMCC) can influence the hydrate formation of theophylline. In particular, the aim was to study if SMCC offers protection against the formation of theophylline monohydrate relative to alpha-lactose monohydrate in wet masses after an overnight equilibration and the stability of final granules during controlled storage. In addition, the aim was to study the use of spectroscopic methods to identify hydrate formation in the formulations containing excipients. Off-line evaluation of materials was performed using X-ray powder diffractometry, near infrared and Raman spectroscopy. alpha-Lactose monohydrate with minimal water absorbing potential was not able to prevent but enhanced hydrate formation of theophylline. Even though SMCC is able to take large amounts of water into its internal structure, it was able to inhibit the formation of theophylline monohydrate only at low moisture contents, not at the amounts of water needed to form granules. Both the spectroscopic methods used could identify the hydrate formation even though there were excipients in the formulation. PMID- 12587114 TI - In situ moisture determination of a cytotoxic compound during process optimization. AB - A simple and safe prototype apparatus was designed and adapted for the in situ determination of the moisture content of a cytotoxic compound (9-fluorenylmethyl protected doxorubicin-peptide conjugate, or Fm-DPC) by near-infrared absorbance spectroscopy during optimization of the chemical isolation procedure. The cytotoxic nature of the compound restricts one's ability to safely sample such drying processes for more traditional means of moisture determination for fear of hazardous solids dusting, hence in situ sampling approaches are of great importance. These concerns also exist for the process development laboratory, where despite the smaller scale of operations, the volume of experiments (hence cytotoxic samples) required to define a chemical process is often more significant. In this application, partial least squares regression was used with Karl Fischer volumetric titration analysis to generate a calibration model. Although pronounced differences in cake density were observed as a function of the buffer selected for the isolation process, the model still achieved a standard error of calibration of 0.63% w/w and a standard error of prediction of 0.99% (w/w). These results demonstrated the versatility of the prototype apparatus/data processing approach to model Fm-DPC drying under extremely variable conditions, as inherently expected during the investigational laboratory development of a chemical process. PMID- 12587115 TI - Formation of physically stable amorphous drugs by milling with Neusilin. AB - Each of four drugs (ketoprofen, indomethacin, naproxen, and progesterone) was milled with Neusilin (amorphous magnesium aluminosilicate) to effect conversion from crystalline to amorphous states, and the physical stability of the resultant drugs was investigated. Ball milling the drugs alone for 48 h resulted in no amorphization. X-ray powder diffractometry (XPD), birefringence, and differential scanning calorimetry (DSC) data indicated amorphization of all the four drugs on ball milling with Neusilin. Fourier transform infrared spectroscopy (FTIR) data showed a reduction in the absorbance of the free and the hydrogen-bonded acid carbonyl peaks accompanied by a corresponding increase in the absorbance of the carboxylate peak, indicating an acid-base reaction between the carboxylic acid containing drugs and Neusilin on milling. On storage of milled powders (at 40 degrees C and 75% RH for 4 weeks), XPD, birefringence, and DSC data showed the absence of reversion to the crystalline state, and FTIR data revealed continued absence of the carbonyl peaks. Whereas the carboxylic acid-containing drugs convert from their crystalline acid form to amorphous salt form on milling with Neusilin, progesterone seems to interact with Neusilin via hydrogen bonding. The amorphous Neusilin-bound states of all four drugs were physically stable during storage. The water of adsorption seems to mediate the conversion of the crystalline state to an amorphous Neusilin-bound state. PMID- 12587116 TI - Multiple pharmacokinetic parameter prediction for a series of cephalosporins. AB - The goal of quantitative structure-pharmacokinetic relationship analyses is to develop useful models that can predict one or more pharmacokinetic properties of a particular compound. In the present study, a multiple-output artificial neural network model was constructed to predict human half-life, renal and total body clearance, fraction excreted in urine, volume of distribution, and fraction bound to plasma proteins for a series of cephalosporins. Descriptors generated solely from drug structure were used as inputs for the model, and the six pharmacokinetic parameters were simultaneously predicted as outputs. The final 10 descriptor model contained sufficient information for successful predictions using both internal and external test compounds. Descriptors were found to contribute to individual pharmacokinetic parameters to differing extents, such that descriptor importance was independent of the relationships between pharmacokinetic parameters. This technique provides the advantage of simultaneous prediction of multiple parameters using information obtained by nonexperimental means, with the potential for use during the early stages of drug development. PMID- 12587117 TI - Measurement and evaluation of the adhesive force between particles by the direct separation method. AB - We developed a new apparatus to measure the adhesive force between particles with a high resolution of approximately 2 nN. The force was measured directly by applying a pulling force to the particles with a contact needle. In addition, the separation process was observed with a 3CCD camera during the measurement. The adhesive force of six kinds of pharmaceutical particles and glass beads was measured, and the effects of moisture, shape, and triboelectrification of the particles on the force were investigated. In the case of moisture-adsorptive particles, the force of adhesion increased rapidly with moisture content under high relative humidity. In the case of moisture-nonadsorptive particles, the force was affected by the particle shape and triboelectrification. The adhesive force of particles having sharp corners was greatly affected by triboelectrification at the corners of the particles. PMID- 12587118 TI - Altered expression and function of P-glycoprotein in dextran sodium sulfate induced colitis in mice. AB - P-glycoprotein (P-gp), the product of the multiple drug resistance (mdr) gene, can actively pump toxic drugs out of cells, but its pathophysiologic role is not yet fully understood. In this study, we examined the expression of P-gp in dextran sodium sulfate (DSS)-induced colitis in mice. Eight-week-old Balb/c female mice were given drinking water containing 7% DSS ad libitum for 7 days. Mice receiving DSS were sacrificed on days 3, 5, and 7 for histopathologic study. Tissue samples were examined by hematoxylin and eosin (HE) staining, and immunostained against mdr, CD4+, CD8+, and B220+. RNA was isolated from the large intestine and the expression of mdr1a was determined by RT-PCR. The function of P gp was evaluated by rhodamine123 efflux using the everted sac method. The induction of colitis in mice was confirmed by body weight changes, HE staining and immunohistological grading of the large intestine with reference to CD4+, CD8+, and B220+ after 7 days of treatment. Severe inflammation was observed in the large, but not the small, intestine on day 7. The expression of mdr1a in the large intestine was reduced on days 3, 5, and 7. In addition, the P-gp function and the expression of PXR were also reduced in the large intestine of DSS-treated mice on day 3. This reduction was consistent with the immunohistologic observations. The expression of the mdr1a gene was reduced before severe symptoms appeared. These results suggest that P-gp expression may be related to the pathology of colitis. PMID- 12587119 TI - Solid-state characterization of a novel chemotherapeutic drug. AB - We present the results of a thermal, spectroscopic, diffractometric, and microscopic study of a novel DNA intercalator synthesized by Novuspharma S.p.A. (code name BRR 2778, purity by high-performance liquid chromatography: 99.4%). We found that the form that is stable at room temperature contains 1.5 water molecules per unit of formula (or about 4.7% in mass): this water is reversibly lost in two stages below 80 degrees and 90 degrees C in dry and wet nitrogen atmosphere, respectively. The hydrated compound is a pseudo-polymorph and dehydration is accompanied by a structural change that modifies the diffraction pattern without changing the shape of the microcrystals. Annealing above 150 degrees C causes decomposition of the anhydrous form and (above 190 degrees C) amorphization of the solid residue occurs. PMID- 12587120 TI - Reaction of a peptide with polyvinylpyrrolidone in the solid state. AB - During stability studies at high temperature (70 degrees C) and low relative humidity ( approximately 0%), the recovery of an asparagine containing hexapeptide (VYPNGA) and its known deamidation products from solid polyvinylpyrrolidone (PVP) matrices was incomplete. To determine the causes of this mass loss, formulations were prepared by lyophilizing solutions containing PVP, glycerol, and the Asn-hexapeptide in pH 7.5 phosphate buffer, followed by storage at 70 degrees C and 0% relative humidity. Asn-hexapeptide loss was mono exponential and reached a plateau at about 30% remaining. Total recovery of the peptide and its known deamidation products was approximately 30% of peptide load. Size exclusion chromatography with fluorescence detection indicated the formation of a PVP-peptide adduct that was stable in the presence of 6 M guanidine hydrochloride. Similar stability studies using N-acetyl phenylalanine, phenylalanine ethyl ester, and N-acetyl tyrosine ethyl ester demonstrated that the reaction involves the peptide N-terminus. The adduct was disrupted in the presence of carboxypeptidase-A, suggesting the formation of an amide bond between the peptide and PVP. (15)N solid-state nuclear magnetic resonance spectroscopy using (15)N-labeled valine as a model of the peptide N-terminus showed different populations of (15)N, suggesting that noncovalent peptide-polymer interactions precede amide bond formation. PMID- 12587121 TI - Extracellular glucose concentration alters functional activity of the intestinal oligopeptide transporter (PepT-1) in Caco-2 cells. AB - The objective of this study was to determine the effect of different cell culture media glucose concentrations on the functional activity of PepT-1 in Caco-2 cells. Uptake kinetics of Gly-Sar into Caco-2 cells that were maintained in iso osmotic media containing 25 or 5.5 mM glucose were determined in the presence and absence of amino acid-selective chemical modifiers and dithiothreitol. Inhibition of Gly-Sar uptake into Caco-2 cells was measured in the presence of dipeptides and xenobiotics exhibiting various binding affinities for the PepT-1. The effect of extracellular glucose on PepT-1 gene expression was assessed using comparative RT-PCR. Long-term exposure of Caco-2 cells to 25 mM glucose reduced maximum transport capacity for Gly-Sar uptake without altering PepT-1 gene expression. In contrast, binding affinity of Gly-Sar and other dipeptides or xenobiotics was not significantly changed. Chemical modification of Lys and Tyr residues decreased V(max), while Cys modification increased the maximum transport capacity of the carrier. Preincubation of Caco-2 cells with dithiothreitol restored PepT-1 activity in cells maintained at 25 mM glucose. In conclusion, cell culture media containing 25 mM glucose decreases maximum transport capacity of PepT-1 in Caco-2 cells without affecting substrate recognition, at least in part, mediated via an oxidative pathway. PMID- 12587122 TI - Variability of intestinal expression of P-glycoprotein in healthy volunteers as described by absorption of talinolol from four bioequivalent tablets. AB - The beta(1)-selective blocker talinolol is incompletely absorbed in man from an "absorption window" in the upper small intestine which is under control of P glycoprotein. The following single dose, four-period, changeover study with 7 days washout in 36 healthy subjects (21 females, age 20-33 years) was designed to confirm bioequivalence of four marketed tablet formulations of talinolol with identical in vitro liberation and to deduce from the intrasubject and intersubject variability of talinolol pharmacokinetics on the variability of intestinal P-gp function. All point estimates of the primary criteria AUC(0 infinity) and C(max) for the comparison of the galenic forms were within 0.9 1.10. The 90% confidence intervals were entirely within the standard ranges of bioequivalence (0.80-1.25 for AUC(0-infinity), 0.70-1.43 for C(max)). The intra- and intersubject coefficients of variation for AUC(0-infinity) were 14.0% and 20.4-29.5%, respectively. In conclusion, the four talinolol tablets are bioequivalent in extent and rate of absorption. The low intrasubject variability of the AUC(0-infinity) after weekly administration of the tablets refers to a small intrasubject variability of the "absorption window" and elimination of talinolol that most likely depends on the expression of P-gp in the small intestine. PMID- 12587123 TI - The effect of mechanical processing on surface stability of pharmaceutical powders: visualization by atomic force microscopy. AB - Atomic force microscopy was used to investigate the influence of mechanical processing (milling) on the surface stability of salbutamol sulfate. Phase imaging, a development of tapping mode atomic force microscopy, was used to elucidate variations in the physico-mechanical properties on the surface of salbutamol sulfate crystals by measuring the phase lag of an oscillating tip in contact with the surface. Simultaneous measurements of topographical and phase images indicated an increase in disorder on the surface as milling time was increased. Specific regions on the milled samples, independent of topography, showed large variations in phase shift (> 30 degrees). These regions (not observed on the crystalline salbutamol sulfate) suggested large differences in the physical properties on the surface. It is therefore reasonable to conclude that these regions were likely attributed to mechanically induced amorphous domains. PMID- 12587124 TI - Classification structure-activity relations (C-SAR) in prediction of human intestinal absorption. AB - AB/HIA is a "soft" filter for identifying compounds with poor intestinal membrane permeability. The analyzed data set included over 1000 drug-like compounds with experimental human intestinal absorption (HIA) values. A sequence of recursive partitioning analyses based on multiple physicochemical and structural descriptors led to the derivation of the rule-based algorithm (filter). The obtained rules reveal a simple physicochemical model of intestinal permeability; they also account for the specific effects caused by quarternary nitrogens and biphosphonate groups. Comparison of the observed and predicted values revealed very low percent of disagreement (15% false-positives and 3% false-negatives). The unusual absorption of compounds that deviated from the predicted values was explained in terms of active transport, efflux, chemical stability, chelating ability, and solubility. Most of these effects can be accounted for by new, substructure-specific rules that can be added into the existing filter. This can lead to the development of a reliable theoretical model for predicting human intestinal absorption. If combined with other models for predicting first pass metabolism, the updated AB/HIA filter can be very useful in predicting oral bioavailability. PMID- 12587125 TI - Separation and characterization of the colloidal phases produced on digestion of common formulation lipids and assessment of their impact on the apparent solubility of selected poorly water-soluble drugs. AB - Colloidal mixtures containing bile salts (BS), phosphatidylcholine (PC), and medium and long-chain monoglycerides and fatty acids were prepared as model systems to represent typical intestinal contents after digestion of formulation derived lipids under both low (5 mM BS/1.25 mM PC) and high (20 mM BS/5 mM PC) BS and PC conditions. Size-exclusion chromatography of the colloidal species that formed in the medium-chain digests indicated the presence of vesicles, mixed micelles, and simple micelles, whereas the long-chain digests contained only vesicles and mixed micelles. In the long-chain digests the mixed micellar phase was the predominant drug solubilizing species for griseofulvin, danazol, and halofantrine, although for increasingly lipophilic drugs, the vesicular phase contributed an increasing proportion of the solubilization capacity. In contrast, the solubilization capacity of the vesicular phase was predominant in the medium chain digests, and no clear trends were evident in the relationship between drug lipophilicity and proportional solubilization. These data highlight the need to consider the colloidal species that form in the small intestine during the digestion of common formulation lipids and the coincident enhancement in drug solubilization provided under these circumstances. PMID- 12587126 TI - Interaction of artemisinin and its related compounds with hydroxypropyl-beta cyclodextrin in solution state: experimental and molecular-modeling studies. AB - Hydroxypropyl-beta-cyclodextrin (HPBCD) was investigated as a possible solubilizer for a series of poorly water-soluble antimalarial drugs. The solubilities of artemisinin, artether, dihydroartemisinin, and 10 deoxoartemisinin in HPBCD solutions were studied. The phase-solubility profile of these drugs in HPBCD solutions, in the concentration range studied, can be classified as type A(L) or soluble 1:1 complexes. The solubilities of artemisinin, artether, dihydroartemisinin, and 10-deoxoartemisinin in 20% w/v solutions of HPBCD are 4.5, 1.3, 6.0, and 5.2 mg/mL, respectively. The stability constants of artemisinin, dihydroartemisinin, artether, and 10-deoxoartemisinin complexes with HPBCD are 475, 405, 327, and 146 M(-1), respectively. Three different docking methods, SYBYL DOCK, FlexiDock, and DOCK 4.0.1 were evaluated to further understand the complexation modes and applicability of the docking programs for the modeling of inclusion complexes. The results showed that DOCK 4.0.1 offers a better correlation in terms of orientation of molecules inside the cyclodextrin cavity and also in terms of docking scores. PMID- 12587127 TI - Prediction of skin penetration using artificial neural network (ANN) modeling. AB - Artificial neural network (ANN) analysis was used to predict the skin permeability of selected xenobiotics. Permeability coefficients (log k(p)) were obtained from various literature sources. A previously reported equation, which was shown to be useful in the prediction of skin permeability, uses the partial charges of the penetrants, their molecular weight, and their calculated octanol water partition coefficient (log K(oct)). The equation was used to predict the skin permeability for the set of 40 compounds (r(2) = 0.672). A successful ANN was developed and the ANN produced log k(p) values that correlated well with the experimental ones(r(2) = 0.997). The penetration properties of a selection of compounds through human skin that have not been previously investigated, etodolac, famotidine, nimesulide, nizatidine, ranitidine, were investigated. Their permeability coefficients were determined. It was then possible to compare the experimental data with that predicted using the partial charge equation and the trained ANN. ANN modeling for predicting skin permeability was found to be useful for predicting skin permeability coefficients of compounds. In conclusion, the developed and described ANN model in this publication does not require any experimental parameters; it could potentially provide useful and precise prediction of skin penetration for new drugs or toxic penetrants. PMID- 12587129 TI - Microencapsulated liposomes in controlled drug delivery: strategies to modulate drug release and eliminate the burst effect. AB - The release of fluorescein isothiocyanate labeled bovine serum albumin (FITC-BSA) from alginate-microencapsulated liposomes was studied to evaluate the properties of this system for controlled drug delivery. Liposomes composed of phosphatidylcholine (PC) and cholesterol (Chol) (molar ratio 7:3) and of PC, phosphatidylglycerol (PG), and cholesterol (6:1:3) were encapsulated in alginate (Alg) crosslinked with Ca(2+) (Ca-Alg), Al(3+) (Al-Alg), and Ba(2+) (Ba-Alg). Capsules were coated with poly(l-ornithine) followed by a final alginate coat. A rapid initial burst of protein release was observed from liposomes encapsulated in Ca-Alg and Al-Alg. No burst was observed when liposomes were encapsulated in Ba-Alg, indicating that the crosslinking ions could significantly affect the release of entrapped protein. Also, the release from encapsulated liposomes varied significantly with liposome composition, especially with Ca-Alg as observed with encapsulation of PC, dioleoylphosphatidylcholine (DOPC), and DOPC/Chol liposomes. Cholesterol increased the leakiness of the liposomes after encapsulation. In all cases, the release from microencapsulated liposomes was much faster than that from free liposomes suggesting an interaction between the liposomes and the alginate. Differential scanning calorimetry supports the hypothesis that alginate was inserted into the lipid bilayer resulting in a rapid release of protein from microencapsulated liposomes. Moreover, it was observed that the degree of interaction between liposomes and alginate varied with liposome composition. PMID- 12587128 TI - Structural stability of adenovirus type 5. AB - Thermally induced structural changes in adenovirus type 5 (Ad) in the presence of either 2 or 10% sucrose were investigated using a variety of biophysical techniques. In solutions containing 2% sucrose, a highly cooperative transition in the structure of the virus was observed at 45 degrees C as detected by tryptophan fluorescence, derivative UV absorption spectroscopy, circular dichroism (CD), and dynamic and static light scattering. This transition resulted in (at least partial) disassembly of the virus and a concomitant increase in the accessibility of the viral DNA to the fluorescent dye, TOTO-1. Capsid disassembly was verified by transmission electron microscopy, which showed ruptured icosahedral vertices near 45 degrees C followed by complete capsid disassembly at higher temperatures. SDS-PAGE of thermally treated Ad suggests that the penton base (protein III) and protein IIIa (located in the peripentonal region) are significantly more labile than other capsid proteins and may be the initial instigators in capsid disassembly. Additional discrete structural transitions were observed in viral proteins using the aforementioned spectroscopic techniques. Thermally induced rearrangements of the condensed DNA at higher temperatures were also detected by the appearance of "psi"-like features in the CD spectra as well as a dramatic decrease in accessibility of DNA to TOTO-1. These transitions corresponded to discrete endothermic events that are also detected by differential scanning calorimetry. By increasing the concentration of sucrose to 10%, secondary and tertiary structural features of adenoviral proteins were significantly stabilized, although loss of quaternary structure at 45 degrees C was still observed. PMID- 12587130 TI - Imagining the developmentally disabled and mentally retarded: an introduction. PMID- 12587131 TI - Respect for the developmentally disabled and forgoing life-sustaining treatment. AB - A developmentally disabled person should be treated at all times as a unique individual and not as some anonymous "disabled person." The developmentally disabled should not be subjected to invasive medical treatment that is unduly burdensome or nonbeneficial, or be forced to endure a quality of life not meaningful to them as individuals. They have a right to refuse or accept treatment that a surrogate must exercise on behalf of each individual in a responsible and careful manner. Three cases and a preliminary approach to the ethical analysis of decisions to allow developmentally disabled persons to die by forgoing medical treatment are offered. The "best interests" of a developmentally disabled individual, properly understood, can serve as a useful and ethically defensible standard for determining the ethical propriety of surrogate decision making about forgoing life-sustaining medical treatment of the disabled. PMID- 12587133 TI - Attitudes, souls, and persons: children with severe neurological impairment. PMID- 12587132 TI - Withdrawing life-sustaining medical treatment--a physician's personal reflection. AB - The decision to withhold or withdraw artificially provided hydration and nutrition is one which evolves over time and must be made jointly by the medical team and the patient's family. Although withholding nutrition can be argued to be ethical and appropriate for certain clinical scenarios, it is still a decision which can be difficult to make and, because of different social and legal issues, can be difficult to carry out. This is the story of one physician's journey as he worked with the mother and father of a young child who suffered a severe neurological injury and was left in a persistent vegetative state. PMID- 12587134 TI - Involuntary sterilization of persons with mental retardation: an ethical analysis. AB - Legitimate concerns on the part of parents and guardians may lead to requests for sterilization of a mentally retarded person in their care. At the same time, mentally retarded persons must be protected from actions that do not serve their best interests. This paper will review the history of involuntary sterilization in the United States and evaluate the ethical arguments that are relevant to decisions about involuntary sterilization. While other, less permanent forms of contraception might be acceptable, involuntary sterilization ought not be performed on mentally retarded persons who retain the capacity for reproductive decision-making, the ability to raise a child, or the capacity to provide valid consent to marriage. Mentally retarded persons who lack capacity in those three areas should be considered for involuntary sterilization only when the procedure is necessary, sterilization would serve the best interests of the mentally retarded person, less intrusive and temporary methods of contraception or control of menstruation are not acceptable alternatives, and procedural safeguards have been implemented to assure a fair decision-making process. PMID- 12587135 TI - Goodness-of-fit ethic for informed consent to research involving adults with mental retardation and developmental disabilities. AB - This article reviews current theory and research on informed consent policies for adults with mental retardation within a relational ethics framework that re conceptualizes consent vulnerability in terms of the goodness-of-fit between participant decisional capacities and the specific consent context. Conceptualizing informed consent competence as a product of the relationship between person and consent context shifts assessment of decisional capacity away from an exclusive focus on a research participant's cognitive deficiencies to (a) an examination of those aspects of the consent setting that are creating or exacerbating consent vulnerability and (b) consideration of how the setting can be modified to produce a consent process that best reflects and protects the hopes, values, concerns, and welfare of adults with developmental disabilities. PMID- 12587136 TI - The ethics of autism: what's wrong with the dominant paradigms and how to fix them. PMID- 12587137 TI - Disability rights critique of prenatal genetic testing: reflections and recommendations. AB - Using prenatal tests to prevent the birth of babies with disabilities seems to be self-evidently good to many people. Even if the testing will not help bring a healthy baby to term this time, it gives prospective parents a chance to try again to conceive. To others, however, prenatal testing looks rather different. If one thinks about the history of our society's treatment of people with disabilities, it is not hard to see why people identified with the disability rights movement might regard such testing as dangerous. For the members of this movement, living with disabling traits need not be detrimental to an individual's prospects of leading a worthwhile life, or to the families in which they grow up, or to society at large. Although the movement has no one position on prenatal diagnosis, many of its adherents believe that public support for prenatal diagnosis and abortion based on disability contravenes the movement's basic philosophy and goals. PMID- 12587138 TI - Ethical aspects of research into the etiology of autism. AB - Advances in understanding autism and other developmental neuropsychiatric disorders will come from an integration of various research strategies including phenomenologic, functional neuroimaging, and pharmacologic methods, as well as epidemiologic approaches aimed at identifying genetic and environmental risk factors. The highly heritable nature of autism makes it scientifically valuable to involve parents and siblings as research participants. However, many studies on autism pose ethical challenges because they do not offer the prospect of direct benefit to subjects. In this article, we present an in-depth ethical analysis of current nontherapeutic research strategies that are common in autism research. The ethical analysis applies a proposed ethical framework for evaluating clinical research focusing on seven ethical requirements: (1) social or scientific value, (2) scientific validity, (3) fair subject selection, (4) favorable risk-benefit ratio, (5) independent review, (6) informed consent, and (7) respect for potential and enrolled research participants. PMID- 12587139 TI - Perinatal substance abuse and human subjects research: are privacy protections adequate? AB - Privacy incursions in the clinical care of substance abusing pregnant women have gained lay and professional attention recently as the result of a high-profile Supreme Court finding in Ferguson vs City of Charleston et al. In March, 2001 the Supreme Court determined that nonconsensual drug screening of pregnant women by clinicians in a public hospital violated the women's Fourth Amendment rights to be secure against unreasonable search and seizure. Coercive or punitive policy approaches to perinatal substance abuse are often based on mistaken assumptions about the nature of addiction and the outcomes of punitive interventions. Much attention has been given to efforts to criminalize pregnancy for drug using women, and civil laws are also coming under increasing scrutiny. Although no state has passed a law criminalizing pregnancy and drug use, an estimated 250 women in more than 30 states have been prosecuted around the country on theories of "fetal abuse." A growing number of states (eighteen to date) have amended their civil child welfare laws to address specifically the subject of a woman's drug use during pregnancy. No one has examined how these laws and social policies could affect research that includes pregnant and parenting women; women (and their families) who stand to lose a great deal should their drug use be brought to the attention of child welfare or criminal justice authorities. We examine the adequacy of current protective mechanisms, such as federal certificates of confidentiality, in protecting research subjects (and investigators) who may be subject to punitive civil or criminal sanctions. We determine that current protective mechanisms may be insufficient to protect research subjects and that investigators and IRB members are often ignorant of the risks imposed by punitive policy approaches to perinatal substance abuse or fall prey to the same mistaken assumptions that inform punitive policies. We conclude that investigators and IRB members have a moral responsibility to understand local, state and national policies and laws governing perinatal substance abuse. Investigators and IRB members should balance the harms of punitive interventions against the protections that may, or may not be afforded to prospective research subjects as well as the prospective benefits, individual and social, of the research. In situations where criminal or punitive policies are in effect, investigators and IRB members should consider whether adequate protections can be achieved. In the context of inadequate protections, potential risks to prospective research subjects and their families may outweigh the individual or social benefits that accrue from the research. Clinical researchers are professionally obligated to work toward amending laws and policies that are not in the best interests of prospective research subjects. PMID- 12587140 TI - Effect of ecdysone agonists on vitellogenesis and the expression of EcR and USP in codling moth (Cydia pomonella). AB - The effects of tebufenozide and methoxyfenozide on vitellogenin (Vg) synthesis/release in the fat body, translocation in hemolymph, uptake by the ovary, and the expression of the ecdysone receptor (EcR) and its heterodimer partner, ultraspiracle protein (USP) in fat body, were investigated in Cydia pomonella. The results indicated that both ecdysone agonists significantly increased the Vg level in the adult hemolymph when the moths were exposed to agonist-treated surfaces. However, these agonists did not affect Vg release from the fat body nor Vg deposition in the first batch oocytes. Western blot analysis revealed that the expression of EcR and USP was significantly increased in tebufenozide- and methoxyfenozide-treated samples compared to the control, suggesting that ecdysone agonists regulated the Vg synthesis via the EcR and USP proteins complex. PMID- 12587141 TI - Small GTP binding proteins: Rab GTPases from the brain of Bombyx mori. AB - From a mRNA of the brain of Bombyx mori, we isolated 8 cDNA clones (BRabs), each of which encodes a different member of Rab-protein family. Four of them have more than 80% amino acid identity to the corresponding members of Drosophila Rab proteins. The other 4 proteins show low sequence similarity to any of the known Rab proteins. However, all of them contain the region conserved in rab protein. Using RACE (Rapid Amplification of cDNA ends), the one full-length cDNA clone (BRab14) was isolated. The clone was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. After purification, the fusion protein was cut with protease to remove GST-Tag and applied to a glutathione S Sepharose column. The protein bound [(3)H]-GDP with association constant of 1.02 x 10(11) M(-1). Further, the protein was phosphorylated by protein kinase. This result suggests that Rab protein in the brain of Bombyx mori binds GDP or GTP and its function is regulated by phosphorylation. PMID- 12587143 TI - Organ-specific distribution of the calcium sensor CaMKII in Locusta migratoria. AB - The Ca(2+)/calmodulin-dependent kinase CaMKII is a key signaling component in Ca(2+)-dependent physiological processes. The expression and function of CaMKII in insect brain is well documented but less investigated for other tissues of insects. The present study demonstrates that in the locust Locusta migratoria CaMKII is widely expressed in various tissues. Relatively high expression levels of CaMKII were found in the brain, upper part of the digestive tract (pharynx, esophagus), and the flight and leg muscles. The different expression patterns of CaMKII in various tissues, as well as different molecular masses of CaMKII between 48 and 60 kDa indicate a tissue-specific expression of CaMKII variants. The expression was monitored with a polyclonal anti-(rat)CaMKII antibody. About 60% of total CaMKII activity in flight muscle cells is associated to the myofibril-rich, particulate fraction suggesting an important role of CaMKII in sarcomeric function. PMID- 12587142 TI - Impact of cysteine proteinase inhibition in midgut fluid and oral secretion on fecundity and pollen consumption of western corn rootworm (Diabrotica virgifera virgifera). AB - Cysteine proteinases predominate in the midgut fluid (MF) and oral secretion (OS) of adult western corn rootworm (WCR) based on their mild acidic pH optima (pH 6.0), enhanced activities after treatment with thiol reducing agents, and inhibition by selective cysteine proteinase inhibitors (PIs). Four cysteine PIs including E-64, calpeptin, calpain inhibitor II, and leupeptin (also a serine PI) strongly inhibited azocaseinolytic activity in a dose-dependent manner in both the MF and OS. The most significant effect on adult female WCR of cysteine PI consumption with corn pollen was the reduction in fecundity, but female survival was not apparently affected. Mean fresh weights for all PI-fed females were also lower than control groups. All PI-fed groups [E-64, calpain inhibitor I (Cal I) and leupeptin] had a significantly lower daily egg production than respective corn pollen-fed controls. E-64 was more potent than leupeptin and Cal I on inhibiting fecundity, which correlates with their relative anti-proteinase potency in vitro. E-64, Cal I, and leupeptin at 1.5-2 nmol/beetle/day reduced fecundity down to 25-45% of control values. Reduced egg production by PI-fed beetles results from a combination of the direct inhibition of protein digestion and a post-ingestive negative feedback mechanism, which reduces food intake. The supplement of ten essential amino acids into the E-64-treated pollen enhanced up to 3.7-fold the number of eggs laid compared to the E-64-fed group without these amino acids, suggesting that egg production is dependent on the supply of essential amino acids from corn pollen proteolysis. PMID- 12587144 TI - Depression and anxiety--separate or continuum? PMID- 12587145 TI - Antidepressant drug discovery in the postgenomic era. AB - The progress made in genome research raises the question whether the new knowledge bases that have emerged may also lead to better antidepressants. The past has seen many remarkable improvements over traditional drugs, but not a real breakthrough. More recently hypothesis-driven research in depression has focussed upon stress-hormone regulation as a possible target, but validation of new drugs is not yet in sight. In parallel, we see an upsurge of systematic unbiased research in a biotechnology-driven drug discovery effort. This research can only lead to results if clinical research adapts to these new demands by phenotyping depressed patients not only according to psychopathological characteristics but also by utilising functional (e.g. neuroendocrine, neuropsychological, neurophysiological, neuroimaging and clinical drug response) data that are to be correlated with data from genotyping. To achieve the goal of genotype/phenotype based differential therapy, large-scale efforts with regards to both patient samples and genotyping capacities are needed. In the long term, increasingly detailed patient information, if translated into specific pharmacological treatments, will lead to customized drugs and thus to a partial fragmentation of the antidepressant market. Concurrently, the improved genotyping/phenotyping efforts will also lead to more widely applicable drugs that promise to avoid side effects and refractoriness and also to hasten the time to onset of action. Once these goals are achieved notorious undertreatment of depression may come to an end. PMID- 12587146 TI - Psychopharmacogenetics--a challenge for pharmacotherapy in psychiatry. AB - Differences in response to treatment or the incidence of adverse drug effects are quite common in clinical psychopharmacotherapy. Although several factors may account for these discrepancies, there is increasing knowledge that genetic factors play a major role. The aim of pharmacogenetics, a new and rapidly growing field in research, is to elucidate the variability in drug response and metabolism due to hereditary differences. According to the hypotheses on the mechanisms of drug action, several mutations in genes coding for neurotransmitter receptors, degrading enzymes, transport proteins or enzymes of the drug metabolizing system (P-450 isoenzymes) have been identified and investigated in psychiatric disorders over the last years. Although some controversy exists among the results, many studies are supportive of the hypothesis that psychopharmacogenetics will be helpful in predicting an individual patient's drug response while minimising the rate of side effects. PMID- 12587147 TI - The activity of beta-hexosaminidase (uHex) and gamma-glutamyl-transferase (uGGT) in urine as non-invasive markers of chronic alcohol abuse: I. Alcohol-dependent subjects. AB - It has been demonstrated that in alcohol-dependent men the uHex test is one of the most powerful discriminating tools, while uGGT has a discriminating power similar to that of sHex but worse than that of uHex, sGGT and s%CDT. Receiver Operating Characteristic (ROC) areas under the curves (AUC) for uHex, uGGT, s%CDT, sHex and sGGT were 0.92, 0.79, 0.88, 0.79 and 0.92, respectively. Due to their good parameters, low cost, ease of use and non-invasive character the uHex and uGGT tests are useful tools for the detection of chronic alcohol abuse. PMID- 12587148 TI - Inositol treatment has no effect on the dexamethasone suppression test. AB - The dexamethasone suppression test (DST) is a widely studied state marker for endogenous depression. Several drugs cause false positives or negatives in this test. Since inositol is a new treatment for depression it is important to determine if it causes artifacts in the DST. Five patients with major depression diagnosed according to DSM-IV underwent a dexamethasone suppression test before and after one and two weeks of 12 grams daily inositol treatment. Three normal subjects underwent the same procedure before and after one week of inositol treatment. Four depressed patients and all three normal subjects demonstrated pretreatment dexamethasone suppression of plasma cortisol. One or two weeks of inositol treatment had no effect on post-dexamethasone cortisol plasma levels in patients or subjects. One depressed patient was a non-suppressor before treatment and continued to show elevated post dexamethasone cortisol levels after one week of inositol treatment. However, after two weeks on inositol, when substantial clinical improvement was noted, he converted to a normal DST. Chronic inositol treatment does not seem to induce false positive DST results. PMID- 12587149 TI - Obsessive compulsive behaviour in autism--towards an autistic-obsessive compulsive syndrome? AB - A portion of autistic patients exhibit compulsive-like behaviours. In addition it has been suggested that serotonin plays a major role in both obsessive compulsive disorder (OCD) and autistic disorder. Other neurohumors such as endogenous opioids and oxytocin have also been implicated in the two disorders. There is also some pharmacological overlap between the two disorders, as well as some similar neuroimaging studies. These similarities and overlaps have led us to propose a putative OCD-autistic disorder, which should be studied in greater detail. PMID- 12587150 TI - Launch of the Cardiac Safety in Schizophrenia Group (CSISG). PMID- 12587151 TI - Response to Dr. S. Curran and Dr. K. Matthew's Letter to the editor (World J Biol Psychiatry 2001, 2: 107) concerning Yaryura-Tobias et al (2000) negative outcome after neurosurgery for refractory obsessive-compulsive spectrum disorder, World J Biol Psychiatry 1: 197-203. PMID- 12587152 TI - A display of hypomania in a depressed male in response to fluvoxamine. AB - The present report describes the behavioural and psychological changes in a 55 year-old depressed male who displayed hypomania after the use of fluvoxamine in addition to other antidepressant medications. The patient experienced his first major depressive episode after the bankruptcy of his company. When fluvoxamine was prescribed at a dose of 50 mg/day in addition to sulpiride at 150 mg/day and a 50 mg dose of trazodone before sleep seven months after admission, grinning and a violation of ward rules occurred repeatedly. The patient became verbally aggressive to the staff and addicted to gambling and alcohol. Six days after the cessation of fluvoxamine, his condition remitted. None of the neuromuscular abnormalities indicative of serotonin syndrome appeared during the episode. Upon review of previous reports on manic switches induced by SSRIs and other antidepressants, we speculate that the fluvoxamine accounted for his hypomania. PMID- 12587153 TI - [Determination of residues of organic chloride pesticide in Chinese medicinal materials with capillary gas chromatography]. AB - Residues of organic chloride pesticide were separated and detected successfully with high performance capillary column and high sensitive electron capture detector, such as hexachloro-cyclohexane, dichloro-diphenyl-trichloroethane, pentachloronitrobenzene. The method can be used for the determination of residues of organic chloride pesticide in Chinese medicinal materials, and the residue enriching in the different part of plant body was tested preliminary. PMID- 12587154 TI - [Studies on calluses induced from various explants of Cistanche deserticola]. AB - Calluses were induced from various explants (flower, stem, bud and squama) of Cistanche deserticola Y.C.Ma and compared with each other. The result showed that the inducement rate of ovary and stem were higher than that of other explants which the inducement time was short and the characters of calluses were good. The effect of basal culture medium added hormone on callus reproduction was also studied. The result showed that B5 culture medium added 1 mg/L IBA and 2 mg/L 6 BA was optimal. PMID- 12587155 TI - [The cluster analysis of trace elements in fructus cnidii from different region]. AB - The contents of trace elements in Fructus Cnidii from different region were assayed by atom absorption spectrum and analyzed by cluster analysis methods. The results showed the contents of trace elements from different regions were different, which had some relativity with habitats, but should be further studied. PMID- 12587156 TI - [Pharmacognostical studies on the root and rhizome of Globba racemosa]. AB - Morphological and histological characters of the root and rhizome of Globba racemosa Smith were described and illustrated with line drawing. These studies provided referential information for the identification and quality control. PMID- 12587157 TI - [Pharmacognostical identification of Polygonum perfoliatum]. AB - In the paper, phamacognostical identification and UV-identification of Polygonum perfoliatum L. were studied. PMID- 12587158 TI - [Parmacognostical identification of Dodartia orientalis]. AB - The descriptions, microsopical characters, UV of Dodartia orientalis L. were studied. It can provide a basis for the identification of D. orientalis. PMID- 12587159 TI - [Study on quality of crude ramulas cinnamomi in different reserve period and their prepared products]. AB - OBJECTIVE: To compare quality of crude Ramulas Cinnamomi in different reserve period and their prepared products. METHOD: Analyzing the content of volatile oil by pharmacopoeia method. RESULTS: The content of volatile oil in prepared Ramulas Cinnamomi by frying decreased, while that in the prepared Ramulas Cinnamomi with honeybee increased a little. The longer the reserve time was, the less the content of volatile oil were. The density becomed smaller in proper order according to crude Ramules Cinnamomi, product processed with honeybee and product processed by frying. But different store time and preparing methods hardly influenced the component of its volatile oil. CONCLUSION: It's useful for Ramulas Cinnamomi to be prepared by frying or with honeybee. PMID- 12587160 TI - [Determination of chemical constituents in the essential oil from Magnolia sprengeri by GC-MS]. AB - The chemical components of the essential oil from Magnolia sprengri Pamp. were analyzed by GC-MS. 30 components composed of about 80% of the total essential oil were separated and identified. PMID- 12587161 TI - [Pharmacological comparison between virosecurinine and securinine]. AB - Pharmacological activities of virosecurinine (Vse) and securinine (Sec) were studied. The results showed that acute toxicity of Vse was 1/13.6 that of Sec, and Vse had no convulsive effects on rats or frogs, while Sec had. The results also showed that Vse and Sec could elevate blood pressure and excite respiration in cats. PMID- 12587162 TI - [The effect of radix pseudostellariae from 8 habitats on spleen-deficiency and immunologic function]. AB - 75% ethanol extracts of Radix Pseudostellariae could improve spleen-deficiency and immunologic function from Yixing(Jiangsu), Tuorong (Fujian), Guangde(Anhui), Zhonggou(Shandong), Linmu(Shandong), Shanghai, Langxi(Anhui) and Xuanzhou (Anhui). They could decrease spleen-deficienly ratio, increase body weight, anus temperature, thymus and spleen indexes, and prolong swimming time in 15 degrees C water and survival time under anoxic circumstances on resperpine-induced mice. They could also inhibit delayed hypersensitivity induced by prednisolone on mice. Radix Pseudostellariae from Xuanzhou was most effective on spleen-deficiency mice. PMID- 12587163 TI - [The effects of safflor yellow on microcirculation and hemorheology of acute blood stasis rats]. AB - OBJECTIVE: To observe the effects of safflor yellow on microcirculation and hemorheology of acute blood stasis rats. METHODS: The acute blood stasis medel was made by i.p. adrenalin (0.07 ml/(kg.time) for 2 times and the stimulation of ice-water. RESULTS: Safflor could markedly improve the number of net dot in microcirculation, and eta b, eta p, eta f in hemorheology (P < 0.05). CONCLUSION: Safflor yellow played an important role on blood stasis of rats. PMID- 12587164 TI - [Effect of "yang wei kang liu" granula on expression of P53 and PCNA in mice bearing sarcoma 180]. AB - OBJECTIVE: To study possible mechanism of "Yang Wei Kang Liu" (YWKL) granula in treating tumor. METHODS: Mice bearing sarcoma180 and immunohisto-chemical method were adopted. RESULTS: Both YWKL granula and its cooperated chemothrapy could restrain the expression of P53 and PCNA. Compared with the chemotherapy group and the cooperated chemotherapy, YWKL granula group was significantly different. CONCLUSION: YWKL granula could restrain the expression of P53 and PCNA, and increase the effect together with chemotherapy. PMID- 12587165 TI - [Determination of forsythin, chlorogenic acid and baicalin in jieduxuantou decoction by RP-HPLC]. AB - RP-HPLC was adopted in determination of forsythin, chlorogenic acid and baicate in Jieduxuantou decoction. PMID- 12587166 TI - [Application of fingerprint analysis and computer pattern interpretation in identification and quality evaluation of traditional Chinese medicines]. AB - The fingerprint analysis technique used in the quality control of TCM through identifying charactreristics and evaluating stability is set up with the rapid development of instrument analyses and computer pattern interpretation. There are many kinds of methods for establishment of fingerpirnt of TCM mainly including chromatography, spectroscopic method, x-ray diffraction and molecular biological technique. The methods of computer pattern interpretation in fingerprint of TCM includes fuzzy information analysis, artificial neural networks and gray relational grade cluster. PMID- 12587168 TI - [Pharmacognostical studies on the rhizome, root and leaf of Alpinia jianganfeng T.L. Wu]. AB - Morphological and histological characters of the rhizome, root and leaf of Alpinia jianganfeng T.L. Wu used as the origin of a traditional Chinese medicine, were described and illustrated with line drawing. These studies provide referential information for the identification and quality control. PMID- 12587167 TI - [Sequences of 5S-rRNA gene spacer region and comparison of alkaloid content in Fritillaria thunbergii from different habitats]. AB - OBJECTIVE: To study the genetic basis of the formation of indigenous Chinese medicine materials. METHODS: The 5S-rRNA gene spacer regions in F. thunbergii from different habitats were amplified with AS and AS-1 as primers, and then sequenced. Total alkaloid contents were assayed by acid dye colorimetry, and 2 main alkaloid contents were assayed by pre-column derivatization and gas chromatographic method. RESULT: The sequenues of 5S-rRNA gene spacer regions in F. thunbergii from different habitats were same, and the length of them was 588 bp. They had same content total alkaloid. The results of gas chromatography showed that they had same kinds of monomer alkaloids, but the contents of different monomer alkaloids were different. CONCLUSION: The difference of alkaloid content in F. thunbergii from various habitats isn't resulted from base sequence variation, but from microenvironment. PMID- 12587169 TI - [The identification of cortex from Kalopanax septemlobus and its confusions]. AB - Descriptions, TLC and UV of cortex from Kalopanax septemlobus and its confusions were carried out to provide a evidence for their identification. PMID- 12587170 TI - [Isolation and identification of two novel polyhydroxylated sterol from Spongia obligue]. AB - Two novel sterols have been isolated from Spongia obligue collected from South China sea and their structure were identified as 24(28)-methylene-1,3,4,11 tetrohydroxycholest-5-en-18-oic acid and 24(28)-methylene-cholestene-5-en 1,3,8,11-tetraol respectively on the basic of the spectral data of MS, IR, 1HNMR and 13CNMR(DEPT). PMID- 12587171 TI - [Determination of phospholipid in Cibotium barometz by supercritical fluid extraction and RP-HPLC]. AB - A supercritical fluid extraction (SFE) method was used to extract phospholipids in Cibotium barometz. After a homogeneous design experiment, SFE conditions were obtained which were pressure at 31.5 MPa, temperature at 46.2 degrees C and a 0.37 L/kg modifier in supercritical fluid. The sample was extracted for 2 hours. A RP-HPLC method was developed for the determination of phosphatidylcholine. RP HPLC conditions included a RP-HPLC column packed with Waters Symmetry C18(5 microns), a mobile phase composed of methanol-1% phoshporic acid(90:10) and the detection wavelength at 206 nm. The content of phosphatidylcholine was 0.198%(RSD 4.2%), and the method can be a measure of quality control for Cibotium barometz and its preparations. PMID- 12587172 TI - [Analysis of essential oil from Artemisia annul L. by extraction of different methods]. AB - Essential oils from Artemisia annul L. were extracted with CO2 supercritical fluid and steam distilation. The essential oils were analyzed and contrasted by GC-MS. 114 components were identified by computer index. The rate of total components by supercritical CO2 fluid extraction and steam distillation were 81.24% and 85.75% respectively. Some of them such as diphenylene-Methane, Phenyl 1-naphthylamine were discovered from the plant for the first time. PMID- 12587173 TI - [The research on the chemical components of Schizonepeta tenuifolia Briq]. AB - Eicosane acid, beta-sitosterol, oleanolic acid, ursolic acid and daucosterol have been firstly isolated and identified from Schizonepeta tenuifolia Briq.. PMID- 12587174 TI - [Protective effect of total flavonoids of fructus choerospondiatis on adriamycin induced rat cardiac peroxidation]. AB - OBJECTIVE: To study the protection effect of total flavnoids of Fructus Choerospondiatis (TFFC) on rat cardiac muscle peroxidation in vitro and vivo. METHODS: Adriamycin-induced peroxidation model on rat was adopted. Activities of LDH, AST and CK in rat's serum, activities of SOD, GSH-Px and content of MDA in rat cardiac muscle were determinated. Activity of LDH and content of MDA in cultured suckling rat cardiac cell were also determinated. RESULTS: Compared with normal group, activities of LDH, AST, CK in serum and content of MDA in cardiac muscle on adriamycin group increased (P < 0.05, P < 0.001), activities of SOD, GSH-Px decreased in cardiac muscle (P < 0.001). Activity of LDH and content of MDA in cultured fluid of suckling rat cardiac cell were higher than that of normal group (P < 0.001). To various dosages TFFC groups, activity of LDH, AST, CK in serum and content of MDA in cardiac muscle decreased; activities of SOD and GSH-Px in cardiac muscle increased; activites of LDH and content of MDA in cultured fluid decreased (P < 0.01, P < 0.001). CONCLUSION: TFFC can maintain the integrity of cytomembrane by scavenging free radicals and inhibiting lipid peroxidation, namely, TFFC has an effect of protecting cardiac muscle. PMID- 12587175 TI - [Antibacterial activity of 20 kinds of Chinese medicinal materials for Helicobacter pylori in vitro]. AB - OBJECTIVE: To study the antibacterial activity of 20 Chinese medicinal materials for helicobacter pylori in vitro and the culture of hylicobacter pylori. METHODS: Doubled-diluted method and tomato juice culture medium were adopted. RESULTS: The detecting ratio of tomato juice culture medium to hylicobacter pylori was equal to that of skirrow culture medium; Radix Scutellariae, Flos Lonicerae, Radix Isatidis, Indigo Naturalis, Fructus Chebulae, Semen Ginkgo, Cortex Phellodendri, Rhizoma Corydalis and Cortex Fraxini have obvious effect of antibacterium to hyliocobater pylori. PMID- 12587176 TI - [Effect of taibai-ginseng on the mice pathology model of "kidney yindeficiency" induced by excessive throxine]. AB - Effect of Taibai-ginseng on mice pathology model of "kidney yindeficiency" induced by excessive throxine was observed in this paper. The result showed taibai-ginseng could improve the pathology symptoms remarkably. It could restrain the rise of body oxygen consuming, body temperature, capacity of drinking water and the decline of pain threshold. PMID- 12587177 TI - [Selection of optimal extraction process for magnolol by orthogonal design]. AB - To find out the optimal extraction process for magnoliae cortex, the extraction process was studied by orthogonal design with yield of the extracta sicca from magnoliae cortex and magnolol content as indexes. Four factors were chosen in this experiment, including alcohol consistence, the alcohol consumption, duration of extraction and times of extraction. The results showed that the optimal extraction process was 65% alcohol consumpted 8 times the amount of magnoliae cortex, refluxing for 3 times, each time 120 min. PMID- 12587178 TI - Empirical methodology as the main criterion of biological psychiatry. PMID- 12587179 TI - ECT has much to offer our patients: it should not be ignored. AB - ECT is an effective treatment for severe mental disorders, including major depression, delusional depression, bipolar disorder, manic delirium, schizophrenia, malignant catatonia, and neuroleptic malignant syndrome. It reduces suicidality, melancholia, catatonia, aggression, and excitement. Age is no barrier, with ECT successful in children, adolescents, and the old-old. It is effective when other treatments have failed. It is a safe treatment. The mortality rate is less than that associated with normal pregnancies. The fractures, panic, and amnesia that marked early treatments have now been modified. Even amnesia, which is often described as the main objection of patients, and the relapse rates are now comparable to the effects of medicines. It is an efficient treatment, costing less for a course of treatment than conventional new medicines and hospitalization. National and international psychiatric societies ignore ECT in their educational programmes. I believe that the leaders have a duty to our patients to provide a forum and educational opportunities for their members for all interventions that may relieve mental disorders. To ignore ECT, an effective treatment, is to do a disservice to our patients, and to abrogate the Hippocratic Oath to which many practitioners subscribe. PMID- 12587180 TI - The neurophysiological meaning of auditory P300 in subtypes of schizophrenia. AB - Event-related potentials are a powerful tool to investigate the real-time course of brain electrical mass activation during cognitive processing. In several psychiatric disorders, differences compared to healthy subjects have been reliably described. The specificity and the pathophysiological meaning of the findings were unclear in most studies, however. This review summarizes methodological aspects and findings, in healthy subjects and psychiatric patients, of investigations based on the auditory oddball paradigm, which evokes the P300 component of event-related potentials. Recent convergent results from P300 and brain imaging studies allowed the interpretation of P300 findings in psychotic disorders in terms of different specific and meaningful neurophysiological disturbances. Namely, core schizophrenia is characterized by a left-temporal dysfunction associated with deficits in verbal processing. Acute remitting schizophrenia-like psychoses (cycloid psychosis, ICD-10 F23), on the other hand, show normal hemispheric balance but consistent signs of cerebral hyperarousal. Recent studies further indicate that the drive for action of manic patients does not rely on over-excitation but rather on frontal disinhibition. The findings may help to further advance the understandings and sub-grouping of functional psychoses based on pathophysiological mechanisms. PMID- 12587181 TI - 50 Years with lithium treatment in affective disorders: present problems and priorities. AB - Lithium has been used as a treatment for various psychiatric- and somatic illnesses for more than 50 years. Today the main use of lithium is for the prevention of episode recurrences in bipolar disorder. The main emphasis of this review will be on the efficacy and effectiveness of lithium prophylaxis in bipolar disorder but the review will also discuss other indications for lithium treatment, the historical development, pharmacokinetic and -dynamic issues, unwanted effects of lithium and the organisation of treatment services. Finally, although not the main purpose of this review, a short description of alternative mood stabilizing drugs will also be presented. PMID- 12587182 TI - Neuroendocrine effects of a short-term osmotic stimulus in patients with chronic schizophrenia. AB - We studied the effects of a short-term hypertonic stimulus on plasma levels of the stress hormones adrenocorticotropin (ACTH), cortisol, prolactin, and the blood volume- and electrolyte-controlling hormones arginine vasopressin (AVP) and atrial natriuretic peptide (ANP). Seven patients suffering from chronic schizophrenia with negative symptoms and ten healthy control subjects were investigated by a 20-minute infusion of 10 ml/kg body weight of hypertonic (2.5%) versus isotonic (0.9%) saline. All patients, who were medication-free for at least one week prior to the study, and all control subjects participated in two investigations in randomized order according to a single-blind cross-over design. During hypertonic infusion, plasma osmolarity and sodium levels were increased similarly in both groups and significantly more than during isotonic saline. Hypertonic saline caused a significant increase of plasma ACTH, cortisol and prolactin in patients in contrast to controls. AVP and ANP plasma concentrations were elevated after infusion of hypertonic saline, however, only patients showed a significant rise in plasma ANP. These results show that a dysregulation of the hypothalamic-pituitary-adrenal (HPA) system in a subset of patients with chronic schizophrenia may become overt during an osmotic stimulation, indicating an increased sensitivity of patients with schizophrenia to osmotic stress. PMID- 12587184 TI - Methodological issues in psychiatry: psychiatry as an empirical science. AB - The paper reflects the empirical position of psychiatry as a research discipline. Principles of empirical research are discussed, such as the generation of empirical hypotheses, the falsification of hypotheses, the logical structure of theories, etc. It is discussed to what extent these principles can be transferred to psychiatric research. PMID- 12587183 TI - Persistent behavioural changes in rats following inescapable shock stress: a potential model of posttraumatic stress disorder. AB - Behavioural changes in rats two weeks after inescapable shock stress were studied using a shuttle-box task (active avoidance/escape). Rats exposed to inescapable shock stress two weeks beforehand showed more frequent avoidance responses and greater within-group variation in both avoidance response and general activity during the test than rats in a control group. General activity during an unstressful period (before starting the test) did not differ significantly between groups. This relatively prolonged increase in responsiveness to external stimuli in the stressful context may be a useful experimental model of posttraumatic stress disorder, especially in relation to hypervigilance. PMID- 12587185 TI - Severe hirsutism associated with psychopharmacological treatment in major depression. AB - We report the case of a 73-year-old female patient with major depression who developed severe hirsutism during six months of psychopharmacological treatment. Old age and polypharmacotherapy appeared to be significant factors that contributed to a sustained disease progression. The higher prevalence of hirsutism in women who suffer from chronic psychiatric diseases highlights the importance of further investigating the aetiology of hirsutism in major depression. PMID- 12587186 TI - Genes for human personality traits: "endophenotypes" of psychiatric disorders? PMID- 12587187 TI - Thyroid hormone, neural tissue and mood modulation. AB - The successful treatment of affective disorders with thyroid hormone exemplifies the suggested inter-relationship between endocrine and neuronal systems in these disorders. Thyroid hormones have a profound influence on behaviour and appear to be capable of modulating the phenotypic expression of major affective illness. Specifically, there is good evidence that triiodothyronine (T3) may accelerate the antidepressant response to tricylic antidepressants, and some studies suggest that T3 may augment the therapeutic response to antidepressants in refractory depressed patients. Open studies have also indicated that adjunctive supraphysiological doses of thyroxine (T4) can ameliorate depressive symptomatology and help stabilize the long-term course of illness in bipolar and unipolar patients, especially women refractory to standard medications. Despite acceptance of the essential role of thyroid hormone on brain maturation and differentiation, and the clinical and therapeutic observations in association with mood disorders, the molecular action that may underlie the mood-modulating properties of thyroid hormone in the adult brain has only recently become the focus of research. The identification of nuclear T3 receptors, the region specific expression of deiodinase isoenzymes and the molecular analyses of thyroid-responsive genes in the adult brain have provided the biological bases for a better understanding of thyroid hormone action in mature neurons. Also the influence of thyroid hormones on the putative neurotransmitter systems that regulate mood and behaviour, serotonin and norepinephrine, may be helpful in explaining their mood-modulating effects. PMID- 12587188 TI - Neurochemical investigation of the schizophrenic brain by in vivo phosphorus magnetic resonance spectroscopy. AB - Abnormal phospholipid metabolisms may play important roles in the pathophysiology of schizophrenia. Phosphorus magnetic resonance spectroscopy (31P-MRS) offers a new method for studying phosphorus-related metabolism in vivo. A decrease in the level of phosphomonoesters (PME) and an increase in the level of phosphodiesters (PDE) has been demonstrated in the prefrontal lobe of neuroleptic-naive schizophrenic patients. Most of the studies in medicated schizophrenic patients have shown decreased PME and/or increased PDE. The decreased PME in the frontal lobe appears to be associated with negative symptoms and poor working memory performance. 1H-decoupled 31P-MRS revealed a reduction in the phosphocholine element of PME and an elevation in the mobile phospholipids of PDE in the prefrontal region of medicated schizophrenic patients. PDE were elevated in the temporal lobes of neuroleptic-naive schizophrenic patients, and this increase was partially normalized by haloperidol administration. Data about the temporal lobes of medicated schizophrenic patients have not been consistent. Except for the reduction in the adenosine triphosphate (ATP) in the basal ganglia and the correlation between the increase in the frontal lobe phosphocreatine (PCr) and negative symptomatology, data related to changes in high-energy phosphates are contradictory. No consensus on the effect of neuroleptics on phosphorus metabolites has been achieved. Methodological problems inherent in 31P-MRS may have contributed to the confusion in understanding available data. Future directions of MRS studies are suggested in the last section of the paper. PMID- 12587189 TI - Structural neuroimaging studies in late-life depression: a review. AB - Which patients presenting with depression in late life will progress to a dementia syndrome has been an important research question in recent times. In this paper we review selectively structural neuroimaging investigations of late life depression (LLD) that have been performed over the past two decades. These studies indicate that there are neuroimaging changes commonly observed in LLD patients when compared to normal controls. Findings include ventricular enlargement and sulcal widening, and reduction in volume size of frontal lobes, hippocampus and caudate nucleus. White matter lesions are more common in depressed subjects and tend to be more severe. Some studies report these changes to be more pronounced in patients who present with late-onset depression (LOD) but this has been contradicted by other studies. Preliminary work suggests that these changes may be associated with a poor prognosis but there is a dearth of systematic, well-controlled longitudinal studies. PMID- 12587190 TI - Alterations of host defence system after sleep deprivation are followed by impaired mood and psychosocial functioning. AB - In healthy humans, sleep deprivation (SD) has consistently been demonstrated to impair different parameters of the host defence system and of psychosocial functioning. However, the individual timing of these alterations and their possible association have remained unknown so far. We therefore investigated functional measures of the individual host defence system as well as of subjective well-being and psychosocial performance in 10 healthy male adults before and after SD, as well as after recovery sleep. In detail, we examined the number of leukocytes, granulocytes, monocytes, lymphocytes, B cells, T cells, T helper and cytotoxic T cells, natural killer (NK) cells as well as the interleukin-1 beta (IL-1 beta) release from platelets after serotonin (5-HT) stimulation. Mood and psychosocial performance (excitement, energy, ability to work and timidity) were measured by visual analogue scales. Taken together, SD induced a deterioration of both mood and ability to work, which was most prominent in the evening after SD, while the maximal alterations of the host defence system could be found twelve hours earlier, i.e., already in the morning following SD. Our findings therefore suggest an SD-induced alteration of these psychoimmune response patterns in healthy humans preceding deterioration of mood and psychosocial functioning. PMID- 12587191 TI - Inositol monophosphatase activity in brain and lymphocyte-derived cell lines of bipolar patients. AB - BACKGROUND: Inositol monophosphatase (IMPase) activity was reported to be low in lymphocyte-derived cell lines of bipolar patients. METHODS: IMPase activity was measured spectrophotometrically as inorganic phosphate liberated from inositol-1 phosphate. RESULTS: The previously reported reduction was replicated in a new, small group of bipolar patients. The reduction is not present in cell lines of unipolar or schizophrenic patients. IMPase activity in postmortem frontal and occipital cortical samples of unipolar, bipolar and schizophrenic patients was not different from controls. CONCLUSIONS: A reduction in lymphocyte-derived IMPase activity without a parallel reduction in cortical IMPase activity could be due to the fact that most leukocyte IMPase activity is the product of the IMPA-2 gene. PMID- 12587192 TI - Movement disorder, memory, psychiatric symptoms and serum DHEA levels in schizophrenic and schizoaffective patients. AB - OBJECTIVE: Reports of low levels of dehydroepiandrosterone (DHEA) or its sulphate (DHEA-S) in some schizophrenic patients and in some persons with poorer motoric and cognitive functioning led us to examine clinical correlates of serum DHEA and DHEA-S levels in schizophrenic patients. METHOD: Ratings of abnormal movements, memory and psychiatric symptoms in 17 medicated chronic schizophrenic or schizoaffective inpatients at a state hospital were correlated with serum DHEA and DHEA-S levels, and their ratios with serum cortisol. RESULTS: Controlling for age, higher DHEA levels and/or higher DHEA/cortisol ratios were significantly correlated with lower symptom ratings on the Brief Psychiatric Rating Scale, better performance on some measures of memory, and lower ratings of parkinsonian symptoms. CONCLUSION: Relatively low DHEA levels or DHEA/cortisol ratios may identify a particularly impaired subgroup of medicated patients with chronic schizophrenia. Potential implications are discussed. PMID- 12587194 TI - Response to Yaryura-Tobias et al (2000) negative outcome after neurosurgery for refractory obsessive-compulsive spectrum disorder, World J Biol Psychiatry 1: 197 203. PMID- 12587193 TI - Attenuation of HPA axis hyperactivity and simultaneous clinical deterioration in a depressed patient treated with mirtazapine. AB - It has been suggested that hypothalamic-pituitary-adrenal (HPA) system dysregulation plays an important role in the pathophysiology of depression and that normalization of HPA axis hyperactivity precedes successful treatment with antidepressants. We report the case of a 61-year-old patient suffering from a major depressive episode who underwent the combined dexamethasone suppression/CRH stimulation test (DEX/CRH test) before and again after one week of mirtazapine treatment. While the patient showed a marked decrease of cortisol and ACTH secretion during the DEX/CRH test within one week, a pronounced and ongoing deterioration of depressive symptoms with suicidal thoughts occurred that was resistant to antidepressant medication and had to be treated with electroconvulsive therapy. Apparently, mirtazapine rapidly attenuates HPA axis hyperactivity in depressed patients via direct pharmacoendocrinological effects. However, this amelioration of HPA system dysregulation is not necessarily accompanied by clinical improvement. PMID- 12587195 TI - Commentary on the nature and treatment of bipolar disorder. PMID- 12587196 TI - Stress hormone-related psychopathology: pathophysiological and treatment implications. AB - Stress is commonly associated with a variety of psychiatric conditions, including major depression, and with chronic medical conditions, including diabetes and insulin resistance. Whether stress causes these conditions is uncertain, but plausible mechanisms exist by which such effects might occur. To the extent stress-induced hormonal alterations (e.g., chronically elevated cortisol levels and lowered dehydroepiandrosterone [DHEA] levels) contribute to psychiatric and medical disease states, manipulations that normalize these hormonal aberrations should prove therapeutic. In this review, we discuss mechanisms by which hormonal imbalance (discussed in the frameworks of "allostatic load" and "anabolic balance") might contribute to illness. We then review certain clinical manifestations of such hormonal imbalances and discuss pharmacological and behavioural treatment strategies aimed at normalizing hormonal output and lessening psychiatric and physical pathology. PMID- 12587197 TI - A serotonin uptake-stimulating tetra-peptide found in urines from ADHD children. AB - A tetra-peptide has been isolated from the urines of children with Attention Deficit Hyperactivity Disorder (ADHD) that we could not find in control urines. The tetra-peptide (G-S-E-N) stimulates the uptake of serotonin into platelets. The peptide may explain why serotonin is increased in platelets of ADHD children. PMID- 12587199 TI - Past, present and future of biological psychiatry. AB - These excerpts from the Presidential Address at the 7th World Congress of Biological Psychiatry, Berlin, 2001 attempt to define the term "biological psychiatry", the principle relevance of diagnostic systems for biological psychiatry and the relevance of biological psychiatry in the past and future for the development of psychiatry in general. They also cover the problem of misuse of biological psychiatry and the need for the rigorous observation of ethical standards. PMID- 12587198 TI - Clinical characteristics of patients with major affective disorders and comorbid migraine. AB - The present study was undertaken to examine the clinical characteristics of patients with major affective disorders and comorbid migraine. Patients (n = 102) with an index episode of either major depression or mania were interviewed with a semi-structured interview based partly on DSM-IV criteria and partly on Akiskal's criteria for affective temperaments. Compared to the patients without migraine (n = 49), the patients with comorbid migraine (n = 53) had a higher frequency of bipolar II disorder (43% vs. 10%), a lower frequency of bipolar I disorder (11% vs. 33%), an approximately equal frequency of unipolar depressive disorder (45% vs. 57%) and a higher frequency of affective temperaments (45% vs. 22%). The migraine patients also had a greater number of anxiety disorders (3.0 vs. 1.9) and a higher frequency of panic disorder and agoraphobia. Gender distribution, age, age at onset of first affective episode, number of previous episodes and symptoms during depressive episodes were similar in both groups. Based on these findings it is suggested that the presence of migraine may be used to delineate a distinct subgroup of the major affective disorders. PMID- 12587200 TI - Acne, isotretinoin treatment and acute depression. AB - The association between isotretinoin therapy and depressive symptoms in acne patients has generated much recent interest but has not been systematically explored. A 17-year-old man with acne vulgaris developed symptoms of acute depression two weeks after beginning isotretinoin therapy. The depressive symptoms improved with reduction of isotretnoin dose and treatment with sertraline. Of note, however, is that when the isotretinoin dose was again increased, the depressive symptoms recurred despite clearing of the skin, leading to an unsuccessful suicide attempt. Isotretinoin was finally discontinued and the depression rapidly resolved. Although the effects of hypervitaminosis A may be involved aetiologically, the predictive factors of drug-related depression remain unclear. Significant depressive symptoms that develop during the course of treatment need close monitoring and may necessitate both antidepressant therapy and discontinuation of the drug. Given the uncertain causal relationship between isotretinoin and depression, versus the potential psychological benefits of effective acne treatment, systematic studies exploring the impact of isotretinoin on mood are needed. PMID- 12587201 TI - [Comprehensive quality appraisal of "maidong"]. AB - According to the contents of 14 kinds of constituents, 6 samples of Maidong from various habitats were comprehensively appraised by the weighted method with factor analysis and fuzzy mathematic method. The results showed that certified Maidong (Ophiopogon japonicus) was superior to its substitute Shanmaidong(Liriope spp.), and the sample from Xiaoshan county was the best in 3 samples of certified goods. Being consistent with the result of the traditional appraisal, the method used in this paper may provide a new means for appraising traditional medicinal materials. PMID- 12587202 TI - [Influence of different collection time on yield and constituents of essential oil of herba pogostemonis from Gaoyao County]. AB - The constituents of the volatile oil of Herba Pogostemonis from Gaoyao county collected at various different time by GC/MS combination technology were analysed and compared. The results showed that the average dry weight of single plant(i.e. yield) increased gradually from July to next April, the content of volatile oil was highest in November. The contents of pogostemone in the leaf oil were higher in July and September, then decreased gradually with growth time; pachouli alcohol was higher in September, October and November, lower in other months. The contents of pogostmone in stem oil were higher in July, October and next January, lowest in November. The content change of pachouli alcohol in stem oil were not obvious in all growth time, higher slightly only after November. Other compounds were not showed their variation law. So the quality of Herba Pogostemonis collected in November was higher if the content of volatile oil was used as a standard of quality evaluation, the result was in accordance with traditional collection experience. PMID- 12587203 TI - [Water purification of breeding hirudo nipponica with photosynthesis bacteria (PBS)]. AB - This paper reported the studies on the water purification of breeding Hirudo nippanica with photosynthesis bacteria (PBS). The results showed the PBS could reduce effectively the chemical dissolved oxygen, NH(4+)-N, vulcanization and organic matter of water bottom, as well as improve the dissolved oxygen. PMID- 12587204 TI - [Identification of radix rubiae and the root of Humulus scardens (Lour.) Merr]. AB - The paper reported the differences of the Radix Rubiae and the root of Humulus scardens (Lour.) Merr. on morphological and histological characteristics and UV identification. PMID- 12587205 TI - [The identification of Hippocampus by HPCE]. AB - Hippocampus from various origins were identified by HPCE. And their character spectrums were established. PMID- 12587206 TI - [Morphological and histological studies of herba sedum aizoon]. AB - The description characters of Herba Sedum aizoon were recounted. And the histological characters of the root, rhizome, stem, leaf and flower of the plant were reported. The characters of the isolated tissue and powder were also involved. These results provided valuable reference information for putting the medicinal resource to rational exploitage and use. PMID- 12587207 TI - [The comparison between regenerated cortex and primary cortex of cortex cinnamomi on description and microscopic characters]. AB - Regenerated cortex of two-year old and three-year old Cortex Cinnamomi were studied on description and microscopic characters and compared with six-year old primary cortex. This provided a evidence for relarging the sources of Cortex Cinnamomi. PMID- 12587208 TI - [Studies on the chemical constituents of the herba of Cuscuta chinensis]. AB - The chemical constituents of the herb of Cuscuta chinensis Lam. were investigated. Five compounds were isolated from petroleum ether and chloroform fraction. Their structures were identified as beta-sitosterol, d-sesamin, 9(R) hydroxy-d-sesamin, d-pinoresinol and daucosterol by chemical and spectroscopical methods. All these compounds were isolated from the stem for the first time. PMID- 12587209 TI - [Study of chemical constituents of essential oil from flowers of Mikania micrantha H.B.K]. AB - The essential oil from flowers of Mikania micrantha H.B.K were extracted and its chemical constituents were analyzed by GC/MS. Forty-two compounds were identified and the main compounds with high contents were beta-cubebene (12.95%), allo aromadendrene (11.67%), beta-caryophyllene (9.17%), 1H-inden-1-one, 5-(1, 1 dimethylethyl)-2, 3-(6.23%), beta-himaohalene (4.56%), trans-alpha-bergamotene (4.09%), limonene (3.68%), beta-ocimene (2.53%). PMID- 12587210 TI - [Studies on the chemical compound of volatile oils from various species of Citrus grandis]. AB - The volatile oils in Citrus grandis from various species were compared by GC-MS. The results showed that the content of volatile oil from Citrus grandis and Citrus grandis (L.) Osbek var. tomentosa Hort. was from 0.7% to 0.9%. Over 30 chemical compounds were identified from the volatile oils and the types of the compounds were almost the same. But gamma-terpinen could not be identified from Citrus grandis which was rich in Citrus gransis (L.) Osbeck var. tomentosa Hort. PMID- 12587211 TI - [Study on inhibitory effect of ampelopsin on melanoma by serologic pharmacological method]. AB - OBJECTIVE: To study the inhibitory effect of ampelopsin on melanoma. METHODS: Using a serologic pharmacological method, the inhibitory effects of different dose of ampelopsin on B16 melanoma cells were evaluated by a MTT assay and a flow cytometry. RESULTS: The serums of C57BL/6 mices, which were administered by 0.420 mmol/kg dose and 0.560 mmol/kg dose of ampelopsin, could obviously suppress the B16 melanoma cells, and the inhibitory rate was equal to that of the vacant serum adding same consistency ampelopsin. The drug serums of the two doses groups reduced the cleavage and proliferation index of B16 melanoma cells to 19.1% and 21.7% respectively. CONCLUSION: Ampelopsin possibly acts on melanoma B16 cells by original form drug at the period with a peak of the drug concentration, and inhibits the DNA synthesis of B16 cells. PMID- 12587212 TI - [Study on antitumor effect and mechanism of aloe polysaccharides]. AB - OBJECTIVE: To study the antitumor activity and mechanism of aloe polysaccharides (AP). METHODS: AP was administered i.p. or i.v. to Sarcoma 180(S180) bearing mice or Hepatoma22(H22) bearing mice solely or combining with CTX, FU and ADM respectively. 10 days later, for S180 mice, the blood was analyzed, the tumor was peeled off and weighed, and the spleen index, thymus index was calculated. For H22 bearing mice, the survival rate was observed or the IL-2, TNF content in serum was tested. RESULTS: 25 mg/kg.d or 50 mg/kg.d AP group could evidently reduce the tumor weight of S180 bearing mice and prolong the survival time of H22 bearing mice. AP also could improve the antitumor effects of CTX, ADM, FU, and lessen the chemotherapy side-effects. Furthermore, AP could improve the level of IL-2, TNF in the serum of mice bearing S180 or H22. CONCLUSION: AP has the effects of antitumor, enhancing the antitumor activity of chemotherapy drugs and lessening their side-effects. This effect was possibly derived from inducing IL-2 and TNF producing in body and improving the immunity activity. PMID- 12587213 TI - [Study on hepatic-protective effect of "yinzhihuang" granula]. AB - OBJECTIVE: To study hepatic-protective effect of "Yinzhihuang" granula. METHODS: By D-GalN, CCl4 induced acute hepatic damage model, ALT and AST in serum, pathological degree of hepar adopted as indexes, the potency of "Yinzhihuang" granula was observed and compared with "Yinzhihuang" oral solutions. Mice administrated ig 1-naphthyl-isothiocyanate, with bilirubin in serum used as index, the effect of "Yinzhihuang" granula was observed. RESULTS: 1.8 g/kg, 3.6 g/kg "Yinzhihuang" granula groups could decrease the rising of ALT, AST in mice serum induced by D-GalN.3.6 g/kg "Yinzhihuang" granula group could also improve pathological degree of hepar.1.0 g/kg, 2.0 g/kg "Yinzhihuang" granula groups could decrease the rising of ALT, AST in rats serum induced by CCl(4).2.0 g/kg "Yinzhihuang" granula group could also improve pathological degree of hepar.1.8 g/kg, 3.6 g/kg "Yinzhihuang" granula groups could decrease the rising of bilirubin in mice serum induced by 1-naphthyl-isothiocyanate. CONCLUSIONS: "Yinzhihuang" granula had an evident hepatic-protective effect on acute hepatic damage induced by D-GalN or CCl4, and could decrease the rising of bilirubin in mice serum induced by 1-naphthyl-isothiocyanate. PMID- 12587215 TI - [Supercentenarians: geography and epidemiology of longevity]. PMID- 12587216 TI - [Neuropathology of the cerebral vessels of centenarians]. AB - Neuropathological study of brain and brain vessels was performed in two series of 12 and 20 centenarians, focusing on the prevalence of small vessel lesions, infarction, Alzheimer's changes and mental status. These are discussed as a function of vascular risk factors. In the first series (12 cases), there was no correlation between the severity of small vessel lesions: hyalinosis (12/12), mineralisation (10/12), amyloid angiopathy (9/12), vascular risk factors (high blood pressure or diabetes), Alzheimer's lesions. However, there was a tendency for an association between amyloid angiopathy and high density of neurofibrillary tangles. In the second series (20 cases), small infarcts and lacunes were found in 9/20 cases, neurofibrillary tangles and diffuse deposits of A beta peptide were constant, senile plaques were very frequent (19/20). Five patients were demented (one vascular dementia, one Alzheimer dementia, and 3 mixed dementias). These data indicate that: 1) Lesions of the walls of small cerebral vessels do not seem linked to the vascular risk factors observed at the end of the life of centenarians. 2) Cerebral infarcts and lacunes are frequent in these patients, and are responsible, at least in part, for a high proportion of the cognitive dysfunctions. The study of larger series is needed for a better understanding of relationships between vascular and degenerative lesions in the oldest old. PMID- 12587217 TI - [Structure and function of the arteries of very elderly people. The PROTEGER Study (Cardiovascular Prognosis and Therapeutic Optimization in Geriatrics)]. AB - The purpose of PROTEGER, a multicenter prospective observational study, was to determine the contribution of hemodynamic, arterial, echocardiographic and biological parameters to the evaluation of individual cardiovascular risk in the elderly. The study included patients aged over 70 years hospitalized in geriatric units with overt cardiovascular disease. Cross sectional analysis of the first 194 subjects included in the study demonstrated a high rate of arterial alterations involving both structure and function. The principal alterations observed were: high pulse pressure despite normal mean systolic and diastolic pressures, frequent and diffuse arterial calcifications, reduced compliance and distensibility, increased thickness, diameter and incremental elastic modulus of the carotid and increased pulse wave velocity. Analysis of monitoring results in the PROTEGER study will demonstrate the role of hemodynamic measurements and arterial alterations in the prediction of cardiovascular risk in hospitalized elderly. PMID- 12587218 TI - [The paradox of hemostasis in the centenarian]. AB - Vascular aging in humans is associated with heightened coagulation enzyme activity. This state of hypercoagulability is compatible with longevity and health. Mechanisms beyond this equilibrium associated with longevity are unknown and studies on polymorphims associated with vascular disease have not been helpful in understanding this phenomenon. PMID- 12587220 TI - Ruth Rice Puffer: highlights of a rewarding life. PMID- 12587219 TI - [Aging and arterial rigidity]. AB - After 50 years of age, the aging process exhibits a characteristic rise in systolic blood pressure associated with a decrease in diastolic blood pressure, resulting in a greater pulsatile pressure in the elderly. This phenomenon is due to increased aortic rigidity and to disturbed wave reflections. These mechanical alterations are modulated by genetic and environmental factors. Drugs may also alter the aging process, increasing longevity as observed over the last 30 years with drug treatment of hypertension. PMID- 12587221 TI - [What is possible, now and right]. PMID- 12587223 TI - The spectrum of accidental childhood poisoning in the Caribbean. AB - OBJECTIVE: To assess accidental poisoning in children in the Caribbean country of Antigua and Barbuda, including the incidence, the types of substances ingested, the age of the children involved, and the clinical outcomes. The results from Antigua and Barbuda were compared with the results of other reports from the English-speaking Caribbean and from the United States of America. DESIGN AND METHODS: We performed a retrospective review of the charts of all patients less than 13 years old admitted to the Children's Ward at Holberton Hospital in Antigua for accidental poisoning between March 1989 and March 1999. Those data were compared with data from earlier reports from Barbados, Guyana, Jamaica, and the United States of America. RESULTS: In Antigua and Barbuda there were 255 hospital admissions for accidental poisoning among children below 13 years old over that 10-year period. Of the 255 ingestions, 115 of them (45%) were in 1-year old children, 69 (27%) were in 2-year-old children, and 26 (10%) were in 3-year old children. These proportions in Antigua and Barbuda are similar to the age patterns seen in the other countries with which we made comparisons. In Antigua and Barbuda there was an annual average of 26 hospital admissions for poisoning for the roughly 20,000 children below 13 years of age, for a rate of 1.3 per 1,000. In comparing the patterns of childhood poisoning in all the countries we studied, we found that, as economic levels rose, there was a shift in the substances ingested, with hydrocarbon and plant ingestions decreasing and chemical and medication ingestions increasing. CONCLUSIONS: There is an increasing variety and complexity of poisonous substances ingested as economic conditions improve. This trend would make the establishment of a poison control center for the English-speaking Caribbean a logical step. PMID- 12587222 TI - [Change in life expectancy in connection with three large groups of causes of death in Medellin, Colombia, between 1989-1991 and 1994-1996]. AB - OBJECTIVES: To examine changes in life expectancy in Medellin, Colombia, between 1989-1991 y 1994-1996, in connection with four large groups of causes of death commonly employed in studies on the burden of disease: group 1, communicable diseases, perinatal and maternal health problems, and nutritional deficits; group 2, non-communicable diseases; group 3, wounds; group 4, ill-defined causes. The latter were excluded from the analysis because of their ambiguity. METHODS: The calculations were made according to the method described by J. H. Pollard in 1986, with the aid of Microsoft Excel, by using the recommended formulas. The computations were double checked with EPIDAT (version 3.0, unofficial). RESULTS: Between 1989-1991 y 1994-1996, a total gain of 1.93 years in life expectancy was seen in Medellin, with a rise from 62.13 to 64.06 years. The gain was greater in men than in women (2.42 vs. 1.09 years, respectively). The increase noted among females was greatest in the extreme age groups (girls 1 to 4 and women over 54 years of age); in men, it was highest in the middle years (between the ages of 25 and 44). In both sexes, the greatest percentage loss in life expectancy was seen in persons 15 to 19 years of age (23% in men and 4% in women, roughly). In the group comprising communicable diseases, perinatal and maternal health problems, and nutritional deficits, a loss in life expectancy was seen in men (0.04 years), whereas in the group of non-communicable diseases there was a gain in life expectancy in both sexes (0.60 years among men and 0.55 years among women). The greater gain in life expectancy among men was linked to a reduction in mortality from wounds (1.98 years). CONCLUSIONS: If one compares the results obtained in Medellin at the end of the study period with life expectancy at birth in Colombia in 1995, which was 70 years, it is obvious that life expectancy in Medellin is still lagging behind, even though it has risen progressively over the years. This is in keeping with the epidemiological transition the city has experienced, which has been linked with a rise in mortality from degenerative and cardiovascular diseases, chronic respiratory ailments, and diabetes, as well as from diseases resulting from human activity. Nevertheless, this epidemiological transition has been slow in Medellin when compared to Colombia as a whole due to high rates of death from infectious and parasitic diseases, which are more characteristic of the transition in its earlier stages. Despite the fact that mortality from wounds has decreased in Medellin, particularly among young males, it may be worthwhile to reassess the effectiveness of interventions undertaken in recent years to promote peaceful coexistence and tolerance in the community. PMID- 12587224 TI - [Endemic goiter prevalence survey in Brazilian schoolchildren 6 to 14 years old, 1994-1996]. AB - OBJECTIVE: To determine the national and state-level prevalence of endemic goiter associated with iodine deficiency in schoolchildren between 6 and 14 years of age in Brazil, based on data from a national survey carried out from 1994 to 1996. MATERIALS: Thyroid inspection and palpation were performed on 178,774 schoolchildren between 6 and 14 years of age, in all the states of Brazil. A subsample was selected to measure the iodine content in their urine as well as the iodine content of the table salt used for cooking in their homes. RESULTS: The last survey before this one was carried out in 1975. We found an 86% decrease in the median prevalence of grade 1 and 2 grade 2 goiter, from 12% to 1%. In 21% of the municipalities, the observed prevalence was zero. From the 16,803 urine samples collected for iodine measurement, 7,702 were matched with the corresponding clinical record (a loss of 54%). The median urinary iodine level was 14.0 micrograms/dL, and without a significant correlation with the clinical data on goiter. The median iodine excretion level for the population in the states of Acre, Amapa, and Tocantins was equal to or below 9.0 micrograms/dL. In three municipalities, urinary iodine was below 2.5 micrograms/dL in all the samples: Parana (in the state of Tocantins), Conceicao (Paraiba), and Nova Roma (Goias). The supplementary iodine found in the salt collected from households (458 samples collected) was below recommended levels, with 50% of them below 20 mg/kg (20 ppm). In 7% of the samples, the level of iodine was below 10 mg/kg of salt, even in salt-producing states such as Rio Grande do Norte. CONCLUSIONS: There is evidence goiter is expanding to parts of Brazil that more recently have been opened up for agriculture, with children in those areas being exposed to low levels of supplementary iodine in the salt used for cooking. We found an increased risk of goiter in areas with deficient iodine supplementation (< 10 mg/kg) (odds ratio = 1.85; Cornfield 95% confidence interval: 1.68-2.03). PMID- 12587225 TI - Adolescents exposed to physical violence in the community: a survey in Brazilian public schools. AB - OBJECTIVE: To determine the proportion of adolescents who have been exposed to physical violence in the city of Porto Alegre, Brazil. METHODS: Fifty-two Porto Alegre public schools that offer basic primary education (grades 1 to 8) were selected through a random sampling process stratified according to school size. In May-November 2000 a screening survey that identifies exposure to violence- experienced personally, witnessed, or knowing victims of violent acts--was administered to all students who were attending a randomly selected 8th grade class in each of the 52 schools. RESULTS: The total number of adolescents included in the analysis was 1,193, representing 10.3% of the students enrolled in 8th grade classes in the city's public school system. On average, each adolescent had been exposed to 19.8 incidents of violence (standard deviation (SD) = 8.5): 2.0 incidents personally experienced (SD = 2.1), 8.5 incidents witnessed (SD = 4.0), and 9.3 knowing a victim of an incident of violence (SD = 4.1). In our sample, the variables of being male (P < 0.0001), being in an older age group (P < 0.001), and not living with both parents (P < 0.001) were independently associated with exposure to more incidents of violence. CONCLUSIONS: We found a high prevalence of incidents of violence among the adolescents in our sample in Porto Alegre. Considering the particular burden inflicted by violent acts on adolescents, health professionals must develop effective approaches to activity identifying, intervening in, and preventing community violence. PMID- 12587226 TI - [Evaluation of a day surgery unit]. AB - OBJECTIVE: To evaluate the Day Surgery Unit (DSU) of the Medical Union of Uruguay Health Center (MUUHC), in Montevideo. The DSU is located in the MUUHC's Sanatorium 1 hospital, a 107-bed hospital that had previously handled low- and medium-complexity surgeries. The results of the reorganization of the surgical activity were analyzed in terms of service utilization, costs, and patient satisfaction. METHODS: For four surgical specialties that were selected for study, health care data were gathered and used to determine the coverage level with the new approach, the number of days of hospitalization, the costs before and after putting the DSU into place, and the patients' opinions of the quality of the care. RESULTS: For the 1998-1999 period, which was the second full year of operation of the DSU, the DSU was carrying out more than a third of the surgeries of the four surgical specialties being studied. These four specialties accounted for 84% of the total number of surgeries carried out by the DSU in the 1998-1999 period. The total number of days of hospitalization declined by 26%. The cost effectiveness analysis showed that the conventional approach, with hospitalization, would have cost some two and a half times as much as the day surgery approach for the same surgeries. In general, the patients expressed strong satisfaction with the services they received from the DSU. DISCUSSION: Our results indicate that it is definitively possible to recommend this day surgery form of care, since it provides good quality of care, it satisfies patients, and it dramatically lessens costs, thanks to the reduction in hospitalizations and the resources needed for inpatient services. It may be possible to expand this new regimen by including other conditions and surgical techniques that are appropriate for day surgeries. PMID- 12587227 TI - [Dental fluorosis: epidemiological fiction?]. AB - OBJECTIVE: To review the scientific literature concerning dental fluorosis and to evaluate its occurrence among children attending public schools in the city of Curitiba, which is in the state of Parana in Brazil. METHODS: We reviewed the international literature on fluorosis that was published between 1998 and 2001, focusing on systematic critical reviews that were listed in such electronic bibliographical sources as MEDLINE, LILACS, the Cochrane Library, and Sci-ELO Public Health. In addition, in 2000 we carried out a cross-sectional study with 12-year old schoolchildren (n = 1,494) in Curitiba. RESULTS: Our literature review found that there is still much controversy regarding the benefits associated with fluoride supplementation and the impact of fluorosis. In our cross-sectional study with the Curitiba schoolchildren, we found a fluorosis prevalence of 23% for grade 2, 3, 4, or 5 on the Dean index. The observed fluorosis had little impact on the biopsychosocial health of the children studied, as shown by the multivariate logistic regression analysis. The analysis showed that the presence of fluorosis was not significantly associated with dissatisfaction with tooth color. However, there was an association between the independent variable of place of residence (sanitary district within Curitiba) and fluorosis (P = 0.00), in both the bivariate and multivariate analyses. CONCLUSIONS: Based on our results, we concluded that dental fluorosis is not now a crucial epidemiological problem for the population studied in Curitiba. Any initiatives to control fluorosis should take into account the population's perception of the problem. Nevertheless, it is still extremely important to monitor the levels of fluoride in drinking water. PMID- 12587228 TI - A comparative analysis of the changes in nursing practice related to health sector reform in five countries of the Americas. AB - OBJECTIVE: To identify changes in nursing practice and the nursing-practice environment that have occurred with implementation of health sector reform in five countries in the Americas. METHODS: An exploratory study of selected settings in Argentina, Brazil, Colombia, Mexico, and the United States of America was conducted between 1997 and 1999 to collect narrative data from 125 professional nurses about their perceptions of nursing practice and changes in work environments. Descriptions of characteristics and trends in nursing practice in the study sites were also obtained. RESULTS: Reorganization of health services has occurred in all five of the countries, responding to health sector reform initiatives and affecting nursing practice in each country. Respondents from all five countries mentioned an emphasis on private enterprise, changes in payment systems for patients and providers, redistributions in the nursing workforce, changes in the personnel mix and nursing-practice functions, work shifting from the hospital to the community, and greater emphasis on cost control and prevention in practice settings. CONCLUSIONS: The study provides initial information about current nursing issues that have arisen as a result of health care reform initiatives. Regardless of differences in service models or phases of health sector reform implementation, in all the countries the participating nurses identified many common themes, trends, and changes in nursing practice. The driving forces for change and their intensity have been different in the five countries. Nurses maintain their core values despite increased work stress and greater patient care needs in all the countries as well as economic crises in the Latin American countries. PMID- 12587229 TI - [Health economics: should it concern the health sector?]. AB - This paper addresses the most important features of health economics, especially its scope and applications within the sphere of health. Health economics is a field of study which allows countries to gear their health policies toward making more rational use of their resources and expanding and improving their health care services. Such policies should, on the other hand, aim to generate strategies for adequately managing human, technical, economic, and financial resources so as to reap the finest health benefits possible. A knowledge of how economics can be applied to health will enable health professionals to introduce an economics culture into their daily work. In other words, it will allow them to keep in mind the scope of their various working tools--health planning, national health accounts and accounting, and economic assessment methods, which include health and pharmaceutical technology assessment and pharmacoeconomics--as well as the place that economics has in health research. Hopefully, a knowledge of these aspects of economic analysis will provide decision-makers with one more tool they can apply in selecting more efficient options and attaining the highest health benefits at the lowest possible cost for the national health system. PMID- 12587230 TI - Policies and managerial guidelines for national cancer control programs. PMID- 12587231 TI - Early assessment of severity in acute pancreatitis. PMID- 12587232 TI - Patients with severe acute pancreatitis should be more often treated in an Intensive Care Department. AB - BACKGROUND: Acute pancreatitis (AP) is a serious disease with a frustrating mortality rate, but with a very good quality of life reported among survivors, that justifies an optimised allocation of therapy intensity. PURPOSE: To audit monitoring and treatment of severe AP in our Intensive Care Department based upon Atlanta severity classification and following recommendations. METHODS: Retrospective study of all AP admitted to our ICU between 1st January, 1993 and 31st December, 1999 in a tertiary University Hospital in Northern Portugal. RESULTS: Our sample (n = 44) represents less than 1% of all patients observed in our ICU and approximately 3% of all patients with AP admitted to our Hospital between 1993 and 1999. All cases fulfilled at least one Atlanta criteria of severe AP. Mean length of stay was 11.6 days. Diagnosis of AP was established in less than 48 hours in 86% of cases: amylasemia and lypasemia were determined in 84 and 7%, respectively and 64% of cases were submitted to ultrasonography. The median time between diagnosis and ICU admission was 2 days. Biliary calculus was responsible for 38% of cases and ethanol for 14%: Thirty-six per cent were considered idiopathic (in none was ERCP performed). Concerning local complications, necrosis was diagnosed in 56% and pseudocysts or abscesses in 23%. Infection was diagnosed by US/CT guided punction or by the presence of gas in CT (performed in 83% during the first ten days of disease) in 18% of the cases. 68% were put on parenteral nutrition (beginning on the 2nd day after admission to ICU in 50% of patients); and 51% had enteric feeding (median day of start = 8.5 days). Antibiotics were prescribed in 91%. Forty-five per cent of patients were submitted to surgery (median day of surgery was 6 days). No statistically significant differences were found concerning local or systemic complications according to different therapies. Mortality rate in our ICU was 36%, mostly during first and second weeks. Patients admitted to ICU later than the second day after diagnosis seem to die earlier (P < 0.005). Outcome (death) was statistically related with organ dysfunction criteria, namely Atlanta criteria (renal failure), SOFA and proportion of days with organ dysfunction. CONCLUSIONS: In our Institution (a tertiary hospital) AP diagnosis is quickly made, local and systemic complications are clearly diagnosed and monitored, but at least 50% of patient waited for 2 days until ICU admission, representing those who die earlier. PMID- 12587233 TI - Bacteriology of anal fistulae. AB - OBJECTIVE: To evaluate by bacteriological analysis the possible relationship between the chronicity of perianal fistulae and the presence of probable permanent infection of the fistulae. MATERIAL AND METHODS: The study included 27 patients, 21 men and 6 women who had undergone surgery for chronic perianal fistulae type II (according to the Parks classification). A total of 27 samples of their fistulous tracts were sent to the microbiology department for the identification of germs. All samples were taken from the theatre and delivered to the laboratory in the same manner. They were then processed under de same conditions in terms of dilutions, inoculation and culture mediums used for the identification of germs in the tracks of the fistulae. RESULTS: A total of 45 were isolated from 21 different species of microorganisms. Most samples (78.26%) had polymicrobic growth. The predominant species were: Escherichia coli (45.45%), Bacteroides fragilis (16.66%), Staphylococcus aureus (12.12%) and Streptococcus viridans (12.12%). No Mycobacteria were found in any specimen. CONCLUSIONS: Anal fistulae can be colonized by poli- or mononormal intestinal or skin microbacterial flora or a combination of both. This report suggests that neither the type or the number of germs its related to the chronicity of cryptoglandular anal fistulae. PMID- 12587234 TI - Villous adenoma of the appendix. Diagnostic and therapeutic approach. AB - Appendix villous adenoma is a rare pathological entity that shows a particular form of clinical presentation that is different from villous adenomas in other places along the gastrointestinal tract. PATIENTS AND METHODS: We reviewed cases of appendix villous adenoma diagnosed in our hospital during the last five years; we studied age, distribution, sex, clinical presentation, diagnostic methods used, treatment and follow-up of patients. RESULTS: We diagnosed 6 cases of villous adenoma of the appendix, acute appendicitis being the most common form of clinical presentation. Treatment was appendectomy in two cases, right hemicolectomy in two patients, subtotal colectomy in one case, and Hartmann's technique with appendectomy in one more case. CONCLUSIONS: We consider appendectomy the treatment of choice when no malignant degeneration is present in the adenoma and invasion of surgical resection borders is absent. PMID- 12587235 TI - Is it a medical error if we do not screen cirrhotic patients for minimal hepatic encephalopathy? AB - Minimal hepatic encephalopathy (MHE) refers to subtle neurocognitive and neurophysiological defects in patients with liver cirrhosis without clinical signs of hepatic encephalopathy. Using appropriate diagnostic methods the prevalence of MHE is approximately 25-30%. MHE has clinical significance as it results in a diminished daily functioning, precedes overt hepatic encephalopathy and is associated with a poor prognosis. Treatment with non-absorbable disaccharides can reverse the neurocognitive and neurophysiological defects found in MHE. The failure to diagnose MHE in apparently normal cirrhotic patients could, therefore, be considered a medical error. However, whether treatment also improves patients' quality of life and prognosis remains to be determined. PMID- 12587236 TI - Retroperitoneal pelvic hemangiopericytoma. Combined abdominoperineal resection. AB - We report a case of large-size pelvic hemangiopericytoma (HPC) incidentally discovered in a 32-year-old woman. Prior to surgery the patient was studied using a CT scan and magnetic resonance imaging (MRI). Because of the size and scarce extent of the tumour (it expanded downwards, into the right ischiorectal fossa) a combined abdomino-perineal approach was needed for complete resection. Seventy two months after initial surgery the patient remains asymptomatic, with no signs of local recurrence or distant metastasis. PMID- 12587237 TI - [Amebic liver abscess]. PMID- 12587238 TI - [Indomethacin-induced diarrhea]. PMID- 12587239 TI - [Esophageal polypoid neoplasm. Carcinosarcoma]. PMID- 12587240 TI - [Reply to "Primary hepatic abscess caused by Actinomyces meyeri" (Rev Esp Enferm Dig 2002; 94(2): 108-9)]. PMID- 12587241 TI - [Reply to "Relation between the cell differentiation degree of colorectal cancer and its topographic distribution". (Linares E, et al. Rev Esp Enferm Dig 2002; 94(2): 78-87) [corrected]. PMID- 12587242 TI - Diagnostic assessment of laryngeal cancer. AB - Laryngeal carcinoma must be diagnosed expeditiously to maximize curative efforts. Our approach combines useful aspects of the patient's presentation with application of appropriate diagnostic modalities. Newer methods of assessment and follow-up are being scientifically validated and will likely enhance the diagnostic efforts of the head and neck surgical oncologist. PMID- 12587243 TI - Imaging in laryngeal cancer: computed tomography, magnetic resonance imaging, positron emission tomography. AB - Imaging modalities have become increasingly significant in the treatment of laryngeal cancer. The ability of CT and MR imaging to reveal pathologic conditions that are undetectable by palpation or endoscopy has led to the development of several imaging-based systems of anatomic classification and tumor staging. This article provides an up-to-date review of the radiographic evaluation of the normal neck and information for both the radiologist and clinician treating the patient with laryngeal cancer. Technical considerations, imaging-based staging systems, and the radiographic characteristics of laryngeal cancer in various regions of the neck are discussed. PMID- 12587244 TI - The molecular biology of laryngeal cancer. AB - Novel techniques have led to the discovery of many genes and gene products important in the development of HNSC and laryngeal cancer. Tumor suppressive genes and oncogenes have been identified, and many of their roles have been elucidated in a genetic progression model. As these molecular pathways become better understood, the information obtained will increasingly be used to guide patient therapy. Specifically, advances will probably be made in (1) molecular characterization of steps leading to laryngeal cancer; (2) molecular screening, staging, and surveillance; (3) molecularly based therapy, including gene transfer and small molecule therapy directed at specific molecular pathways involved in neoplasia; and (4) characterization of patients who are at high risk for laryngeal cancer. In the final analysis, however, smoking cessation for those at risk for head and neck cancer would have greater effect than all these efforts combined. PMID- 12587245 TI - Definitive radiotherapy for squamous carcinoma of the larynx. AB - Definitive radiotherapy for T1 or T2 laryngeal carcinoma offers good local control and quality of voice. Recent clinical trials with hyperfractionated radiotherapy and concurrent chemoradiotherapy demonstrated improved rates of local control for advanced laryngeal cancer treated with primary radiotherapy and provides the opportunity for organ preservation. The use of amifostine and pilocarpine may prevent or ameliorate radiation-induced xerostomia. Recent advances in 3-D conformal radiotherapy and IMRT lead to a better dose distribution for sparing normal organs while treating target volumes with full dose. PMID- 12587246 TI - Organ preservation trials for laryngeal cancer. AB - Organ-preservation strategies include definitive radiation therapy alone, induction chemotherapy followed by radiotherapy, and concurrent chemoradiotherapy. Over the past decade, induction chemotherapy followed by radiotherapy has been the standard for the nonsurgical management of advanced laryngeal cancer. Over this same period, however, other nonsurgical strategies have been under evaluation. These approaches include radiotherapy with concurrent chemotherapy to take advantage of the radiosensitizing properties of most cytotoxic drugs with activity against squamous cell cancer, altered fractionation radiotherapy, and the incorporation of molecularly targeted therapeutics into multimodality treatment. PMID- 12587247 TI - Minimally invasive techniques for management of early glottic cancer. AB - This article reviews management of early laryngeal squamous cell carcinoma with respect to current standards of practice and recent advances in minimally invasive endoscopic surgery, specifically highlighting powered instrumentation as an alternative to transoral laser resection of early glottic lesions. Limitations of the current staging system, controversy regarding management of lesions involving the anterior commissure, and the role of radiation therapy versus surgical management are also discussed with reference to current outcomes literature. PMID- 12587248 TI - Organ preservation surgery for laryngeal cancer. AB - The open organ preservation surgical procedures are an important part of the head and neck surgeon's armamentarium for treating laryngeal cancer. The principles of organ preservation surgery as they apply to laryngeal cancer must be thoroughly appreciated and strictly applied for oncologic and functional success. The selection of eligible patients for these procedures is an art and requires a keen clinical acumen. The vertical partial laryngectomy and supraglottic laryngectomy have defined clinical applications that are relatively well accepted. The supracricoid laryngectomy continues gaining acceptance as a means of treating more extensive glottic and transglottic lesions while maintaining physiologic speech and swallowing without the need for a permanent tracheostoma. The inability to include and use the open surgical organ preservation approaches in the organ preservation paradigm for larynx cancer severely limits the patient's treatment options. Total laryngectomy and medical organ preservation protocols may not be acceptable to the patient from a quality-of-life standpoint. Therefore, it is incumbent upon the head and neck surgeon to have a thorough understanding of all the options available for treatment in the organ preservation paradigm for laryngeal cancer. These options must be skillfully evaluated as they relate to the patient's disease process and confidently used to provide the best oncologic and functional outcome. PMID- 12587249 TI - Total laryngectomy with tracheoesophageal conduit. AB - Tracheoesophageal puncture has made high-quality voice restoration readily available to most individuals who must undergo TL. Although none of these patients would voluntarily choose to undergo TL if not faced with a life threatening condition, at least some achieve a better quality of life, including better vocal function, after treatment and rehabilitation. The methods involved in placement and maintenance of a tracheoesophageal prosthesis are straightforward and permit successful, long-term voice restoration for most patients. Although far from perfect, TL with TEP affords both quality and quantity of life to a group of individuals who currently have no alternative but to undergo debilitating surgery in an effort to extend their lives. PMID- 12587250 TI - Laryngopharyngectomy with reconstruction. AB - A wide range of reconstructive options allows the ablative surgeon to resect tumors completely with wide margins. Wide resection is especially important because of the rich lymphatic drainage and submucosal spread seen with carcinomas in the hypopharyngeal area. Postoperative stenosis can be a difficult, recurring problem if the neopharynx does not have enough tissue incorporated into the closure. Therefore, most laryngopharyngectomy procedures benefit from the addition of transposed tissue, either pedicled or using free tissue transfer microvascular techniques. Often the location of the tumor is a major factor in determining which reconstruction is best for the patient. Minimizing the donor site morbidity must be carefully considered, also. PMID- 12587251 TI - Swallowing and speech therapy after definitive treatment for laryngeal cancer. AB - As the trend in laryngeal cancer treatment shifts towards organ-conservation surgeries and organ-preservation protocols, patients will more often retain anatomy vital to communication and swallowing. Despite a conservative approach, results of treatment may have debilitating effects. Rehabilitation efforts are directed towards a return to functional, if not normal, status. Although there are predictable trends in voice and swallowing disorders of patients with laryngeal cancer, posttreatment dysphonia and dysphagia are diverse in presentation. Considering the significant diversity of this population, speech pathologists should work closely with otolaryngologists to determine the most appropriate treatment for each patient. As this article demonstrates, voice and swallowing therapy are necessary components of the rehabilitation process following treatment for head and neck cancers. As always, treatment is tailored to the specific individual and based on information obtained during a thorough evaluation by a speech pathologist. Fortunately, with the help of voice and swallowing therapy, many patients return to functional communication and oral feeding. PMID- 12587252 TI - Toxicokinetics and physiologically based toxicokinetics in toxicology and risk assessment. AB - Toxicokinetics is the study of kinetics of absorption, distribution, metabolism, and excretion of a xenobiotic under the conditions of toxicity evaluation. Conventional toxicokinetics uses the hypothetical compartments, and the model is composed of rate equations that describe the time course of drug and chemical disposition. The utility of toxicokinetics in toxicity evaluation and interpretation of animal toxicology data is emerging as an important tool in product discovery and development. With implementation of the International Conference on Harmonization (ICH) guidelines on systemic exposure and dose selection, toxicokinetics have been integrated in routine toxicity evaluations. Although traditional compartmental/noncompartmental models are generally adequate for assessing systemic exposure, they are unable to the predict time course of drug disposition in target tissues and often fail to relate systemic drug levels to a biological response. Physiologically based toxicokinetic (PB-TK) models address this deficiency of traditional compartmental models. PB-TK models are the kinetic models of the uptake and disposition of chemicals based on rates of biochemical reactions, physiological and anatomical characteristics. These models, when developed appropriately, can predict target organ drug distribution in different species under variety of conditions. This minireview discusses the basic principles, and applications of traditional compartmental toxicokinetic and physiologically based toxicokinetics (PB-TK) models in drug development and risk assessment. Special emphasis will be placed on discussion related to interpretation of the ICH guidelines related to toxicokinetics and the utility of toxicokinetics data in dose selection for toxicity and carcinogenicity studies. The utility of PB-TK models in risk assessment of methylene chloride, vinyl chloride, retinoic acid, dioxin, and inhaled organic esters is discussed. PMID- 12587253 TI - Combined exposure to DEET (N,N-diethyl-m-toluamide) and permethrin: pharmacokinetics and toxicological effects. AB - Permethrin and DEET are concurrently used for pests control inside homes, in public places, and in military shelters. Combined exposure to these compounds produced greater biochemical, behavioral, and metabolic alterations in animals compared to each individual compound. Concurrent application of DEET and permethrin induced urinary excretion of 3-nitrotyrosine and 8-hydroxy-2' deoxyguanosine, markers of DNA damage and oxidative stress in rats, increased the release of rat brain mitochondrial cytochrome c, disrupted the blood-brain barrier (BBB) in rats, decreased m2 muscarinic acetylcholine receptor ligand binding density in rat brain, increased urinary excretion of 6 beta hydroxycortisol, a marker CYP3A4 induction, altered sensorimotor and locomotor activities in rats, and changed in vivo and in vitro metabolism and pharmacokinetic profiles of the individual compound. These findings show that more research is needed to examine adverse effects of the combined use of DEET and permethrin on other biochemical/physiological system(s) and to predict mechanistic pathways for these effects, particularly mechanism of action at cellular and molecular levels and alterations of genes transcription. PMID- 12587254 TI - 1,3-Butadiene: exposure estimation, hazard characterization, and exposure response analysis. AB - 1,3-Butadiene has been assessed as a Priority Substance under the Canadian Environmental Protection Act. The general population in Canada is exposed to 1,3 butadiene primarily through ambient air. Inhaled 1,3-butadiene is carcinogenic in both mice and rats, inducing tumors at multiple sites at all concentrations tested in all identified studies. In addition, 1,3-butadiene is genotoxic in both somatic and germ cells of rodents. It also induces adverse effects in the reproductive organs of female mice at relatively low concentrations. The greater sensitivity in mice than in rats to induction of these effects by 1,3-butadiene is likely related to species differences in metabolism to active epoxide metabolites. Exposure to 1,3-butadiene in the occupational environment has been associated with the induction of leukemia; there is also some limited evidence that 1,3-butadiene is genotoxic in exposed workers. Therefore, in view of the weight of evidence of available epidemiological and toxicological data, 1,3 butadiene is considered highly likely to be carcinogenic, and likely to be genotoxic, in humans. Estimates of the potency of butadiene to induce cancer have been derived on the basis of both epidemiological investigation and bioassays in mice and rats. Potencies to induce ovarian effects have been estimated on the basis of studies in mice. Uncertainties have been delineated, and, while there are clear species differences in metabolism, estimates of potency to induce effects are considered justifiably conservative in view of the likely variability in metabolism across the population related to genetic polymorphism for enzymes for the critical metabolic pathway. PMID- 12587256 TI - Standards for privacy of individually identifiable health information. PMID- 12587255 TI - Inhaled formaldehyde: exposure estimation, hazard characterization, and exposure response analysis. AB - Formaldehyde has been assessed as a Priority Substance under the Canadian Environmental Protection Act. Probabilistic estimates of exposure of the general population in Canada to formaldehyde in ambient and indoor air are presented. Critical health effects include sensory irritation and the potential to induce tumors in the upper respiratory tract (the nasal region in rodents and potentially the lungs of humans). The majority of the general population is exposed to airborne concentrations of formaldehyde less than those typically associated with sensory irritation (i.e., 0.1 mg/m3). Based primarily upon data derived from laboratory studies, the inhalation of formaldehyde under conditions that induce cytotoxicity and sustained regenerative proliferation within the respiratory tract is considered to present a carcinogenic hazard to humans. At airborne levels for which the prevalence of sensory irritation is minimal (i.e., 0.1 mg/m3), risks of respiratory-tract cancers for the general population estimated on the basis of a biologically motivated case-specific model are exceedingly low. This biologically motivated case-specific model incorporates two stage clonal expansion and is supported by dosimetry calculations from computational fluid dynamics analyses of formaldehyde flux in various regions of the nose and single-path modeling for the lower respiratory tract. The degree of confidence in the underlying database and uncertainties in estimates of exposure and in characterization of hazard and dose response are delineated. PMID- 12587257 TI - Behavioral medicine in the prevention and treatment of cardiovascular disease. AB - Cardiac behavioral medicine is the application of behavioral and psychosocial principles to the prevention and treatment of heart disease. Most biomedical cardiovascular risk factors (e.g., high blood lipids, high blood pressure, diabetes) require behavioral and medical interventions. Other risks, including obesity, high-fat eating pattern, smoking, and inactivity, clearly require lifestyle change. Behavioral medicine screening and intervention have been applied to psychosocial risk factors such as depression, hostility, and social isolation. Appropriate assessment of risk factors is essential because research has demonstrated successful prevention of heart disease and reduction of morbidity and mortality in patients with existing disease. Behavioral interventions have been beneficial in improving cardiac outcomes by enhancing compliance with medication taking and dietary/exercise recommendations. Future needs include the study of psychosocial factors in women and ethnic minorities with heart disease and the integration of behavioral medicine with newer medical technologies designed to detect subclinical biomarkers of heart disease. PMID- 12587258 TI - Assessing depression in the cardiac patient. When is the appropriate time to assess depression in the patient undergoing coronary revascularization? AB - Depression is a well-established risk factor for cardiovascular disease-related morbidity and mortality. It is common to screen for depression in patients undergoing coronary revascularization prior to revascularization; however, the validity of this assessment is unclear as some patients may experience transient, reactive depression rather than persistent depression. The authors evaluated whether an initial or 1-month postprocedure screen was optimal for identifying consistently depressed patients. Depression at 1-month postprocedure was a stronger predictor of depression at months 2 to 6 than baseline depression. After adjusting potential confounding variables, there was a much stronger relationship between 1-month and 6-month depression status (OR = 28.7 if depressed at 1 month, p < .001) than between baseline and 6-month depression status (OR = 6.5 if depressed at baseline, p < .001). Screening for depression at the time of revascularization is not as predictive of depression at 6 months as it is 1 month postprocedure. PMID- 12587259 TI - Program participation and physical activity maintenance after cardiac rehabilitation. AB - This study examined exercise maintenance among patients after completing cardiac rehabilitation. Subjects were men and women who had completed a Phase II Cardiac Rehabilitation Program approximately 12 months previously. Subjects were classified according to whether they had (a) never participated in a Phase III program (G-I) (N = 37), (b) enrolled and completed a Phase III program (G-II) (N = 30), or (c) were currently enrolled in a Phase III maintenance program (G-III) (N = 33). Subjects were significantly more likely to be participating in regular exercise if they had participated in a Phase III program (p < .05). Individuals in G-II and G-III engaged in more minutes of physical activity per week and were more likely to meet recommended levels of physical activity compared to G-1 subjects. G-1 individuals who had longer Phase II programs were more likely to maintain their exercise habits following graduation (p < .05). Results suggest that Phase III maintenance programs and longer Phase II participation improved exercise maintenance following rehabilitation. PMID- 12587260 TI - Socioeconomic status variables predict cardiovascular disease risk factors and prospective mortality risk among women with chest pain. The WISE Study. AB - This study examined the relationship between socioeconomic status (SES), coronary artery disease (CAD) risk factors, and all-cause mortality in a cohort of women with chest pain. A total of 743 women (mean age = 59.6 years) with chest pain who were referred for coronary angiography completed a diagnostic protocol including CAD risk factor assessment, ischemic testing, psychosocial testing, and queries of SES. Patients were followed for about 2 years to track subsequent all-cause mortality. Results indicated that low SES was associated with CAD risk factors, including higher BMI and waist-hip ratios, cigarette smoking, lower reported activity levels, and a greater probability of hypertension. Low income also predicted all-cause mortality (RR = 2.7, 95% CI 1.4, 5.2), including after adjusting for proposed psychosocial and behavioral variables (RR = 5.9, 95% CI 1.2-29.7). Future research will require a thorough a priori focus on potential mechanisms to better understand SES effects on health. PMID- 12587261 TI - Neurocognitive sequelae following coronary artery bypass graft. A research agenda for behavioral scientists. AB - Several studies have demonstrated that a sizeable proportion of patients undergoing coronary artery bypass graft (CABG) demonstrate persistent declines in cognitive functioning. However, several important questions remain regarding cognitive changes following CABG. First, can patients vulnerable to cognitive decline after CABG be identified, providing valuable information that can be factored into clinical decisions? Second, the specificity of CABG as a cause of cognitive decline, when compared to other coronary procedures such as percutaneous coronary interventions, has not been established. Third, what mechanisms account for the neurocognitive decline after CABG? Several mechanisms have been proposed to lead to post-CABG neurocognitive deficits, including pre CABG neurocognitive deficits, physiological injury, psychosocial factors, and/or the patient's perceptual processes. Finally, no study has demonstrated that cognitive changes, as measured by neuropsychological tests, have ecological validity. Specifically, behavioral sciences research is needed to demonstrate that measured cognitive changes significantly impact health status independent of other risk factors. PMID- 12587262 TI - Negative emotion and coronary heart disease. A review. AB - This article reviews literature regarding the influence of negative emotions, specifically depression, anger/hostility, and anxiety on coronary heart disease (CHD). For each domain, evidence is presented demonstrating the deleterious effects of negative affect on health outcomes in patients with CHD. This is followed by a discussion of the manner in which emotional factors are transduced into cardiac health risk factors. The pathophysiological mechanisms by which negative emotions have been found to exert an influence on CHD are highlighted. Finally, a general overview of the outcomes of interventions designed to ameliorate the effects of these negative emotional states on cardiovascular health are reviewed. Several treatment studies are described in detail for the purpose of elaborating the types of multicomponent interventions that attempt to address negative emotions in populations with CHD. PMID- 12587263 TI - Maintenance of health behavior change in preventive cardiology. Internalization and self-regulation of new behaviors. AB - Long-term health behavior maintenance remains a challenge for patients and health behavior interventionists. Resource-intensive systems of external reinforcement and behavioral cues can support behavior maintenance; an alternative approach is to promote patient internalization and self-regulation of health behaviors. Based in part on organismic internalization theory, self-determination theory, and the experience of patients successful at maintaining health behaviors, the health behavior internalization model (HBIM) is proposed to describe motivational factors associated with internalization processes and hypothesizes that integrated internalization may be associated with long-term health behavior maintenance. The HBIM identifies four self-needs (ownership, self-determination, security, and support) and four behavior-related needs (preference, context, competence, and coping) as motivating health behavior internalization. Behavior change strategies promoting integrated internalization are identified from self determination theory, motivational interviewing, and transtheoretical model interventions. Other health behavior change constructs are reviewed in relation to internalization processes, and potential limits to the model are discussed. PMID- 12587264 TI - [Neuroprotective effects of semax in MPTP-induced disturbances of brain dopamine system]. AB - Effects of an ACTH (4-10) analogue Semax (MEHFPGP) on behaviour of white rats with MPTP-induced disturbances of brain DA-system have been studied. It was shown that MPTP administration (25 mg/kg) reduced motor activity and auhmented the anxiety level in rats. Semax administration (daily intranasal 0.2 mg/kg) attenuated behaviour disturbances induced by neurotoxin. The observed protective action of Semax in rats with MFTP-induced DA system disturbances may be due to both its modulating influence on the brain DA system and peptide neuroprotective effects. PMID- 12587265 TI - [C-fos gene expression in the rat spinal cord and brain cells during stress and the use of different types of halothane anesthesia]. AB - C-fos gene expression was studied as a marker of nervous cells activation in the rat spinal cord and brain cells under different conditions (different kinds of narcosis). Using of 1.5% light halothane narcosis allowed the detection of c-Fos like proteins expression in the spinal cord cells only. Under initial 1.5% halothane narcosis, c-Fos-like proteins expression in the rat spinal cord (lumbar segments) and the brain cells was observed after placing the rats into the hammock, noxious mechanical stimulation (NMS) or high frequency electromagnetic irradiation of the skin (EHF). The pattern of the brain structures reacting on the NMS by c-Fos proteins expression, was determined. It was shown that NMS increases the c-Fos positive cell quantity in the lateral hypothalamic area (LHA), ventro-medial (VMH), dorso-medial (DMH) hypothalamic nuclei and anterior hypothalamic area (AHN) by 116, 199, 101 and 157% resp., in comparison with the c Fos immunoreactive cell quantity in intact animals. EHF irradiation of the skin decreased the intensity of c-Fos-like proteins synthesis induced by NMS in the most of the investigated structures (LHA, VMH, DMH and AHA by 32.8, 29, 15 and 33%, resp.). It was shown that only initial halothane narcosis allowed to determine the hypothalamic structures which realized the responses to the NMS, and modification effects of EHF skin irradiation on the intensity of these responses. PMID- 12587266 TI - [Correlation between cataleptic freezing and prestimulation inhibition of the startle reflex in rats]. AB - In rats of GC strain bred for predisdposition to cataleptic freezing, a significant negative correlation between the duration of freezing and the level of prepulse inhibition (PPI) of the startle reflex, has been found. Besides, in a group of GC rats specific by their "nervousness" and jumpiness, there was also a negative correlation between the duration of freezing and the habituation to the startle reflex. None of this correlation have been found in Wistar rats. Since impairment of the PPI and habituation of the startle reflex is considered to be characteristic of schizophrenia, it is believed than cataleptic freezing in the GC rats may be used as a model of schizophrenic psychopathology. PMID- 12587267 TI - [Activation of dopaminergic system stimulates an immune response in mice with opposite type of behaviour]. AB - It was shown that activation of dopaminergic (Daergic) system induced an increase of the immune responsiveness independent of the CBA mice behaviour typeanimals without experience of victories and defeats (control), with aggression and submission. Administration of SKF-38393, a selective agonist of DA D1-receptors, resulted in enhanced immune response as tested by plaque-forming cells and rosette-forming cells number. Similar immunostimulation was observed after injection of p-chlorophenylalanine realizing its influence on the immune response through DA D2-receptors as shown by us elsewhere. It was suggested that activation of Da-ergic system produces a new neurochemical pattern (Daergic neurochemical set) which are responsible for character and intensity changes of the immune response in mice with alternative form of social behaviour. PMID- 12587268 TI - [Effect of selective protein kinase C inhibitor on synaptic plasticity of command neurons of defensive behaviour during sensitization in snails]. AB - Polymyxin B (proteinkinase C inhibitor) effects on nociceptive sensitization of semiintact preparation were investigated in LP11 and RP11 snail neurons. It was found that application of sensitizing stimuli to control snail head initiated neural membrane depolarization, increase its excitability as well as depression of neural responses evoked by sensory stimulation during short-term stage. Polymyxin B application suppressed neural responses evoked by sensitizing (nociceptive) stimuli. At the same time changes in neural membrane excitability as well as neural responses evoked by tactile stimulation of snail foot or chemical stimulation of snail head were similar with ones in control snails. Polymyxin Bdid does not change the depression of neural responses evoked by tactile stimulation of snail head during short-term stages of sensitization but significantly suppressed facilitation of neural responses evoked by tactile stimulation of snail head during long-term stage of sensitization. It was suggested that proteinkinase C is involved in regulation of nociceptive mechanisms as well as in plasticity selective induction mechanisms in command neuron synaptic inputs activated by tactile stimulation of snail head. PMID- 12587269 TI - [Localization of motor and speech cortex in the brain with functional magnetic resonance tomography]. AB - Functional magnetic resonance imaging (MRI) can be used for non-invasive mapping of cerebral cortex. The purpose of the study was evaluation of applicability of functional MRI for studies of neurosurgical patients. 32 volunteers (mean age 37.8 +/- 20.9 years) and 16 patients with brain tumors (mean age 36.2 +/- 24.2 years) were included in the study. Statistical analysis of the data obtained was performed. Activation maps were superimposed on anatomical images and discussed with neurosurgeons. Functional MRI studies were successful in localising the motor cortex and Broca's area in 89% of cases. In 69% of cases, results of the functional MRI influenced the patients' treatment. PMID- 12587270 TI - [Function of hypothalamic-pituitary-adrenocortical system in hypertensive rats of ISIAH strain]. AB - Functional activity of hypothalamic-pituitary-adrenocortical axis has been studied under control and restraint stress conditions in rats with inherited stress-sensitive arterial hypertension (ISIAH strain) and in normotensive WAG (Wistar Albino Glaxo) strain. The levels of hypothalamic CRH-mRNA (in control and 2 hrs stress), pituitary and plasma ACTH and plasma corticosterone (in control and after 5, 15 or 30 min of restraint stress), were evaluated. Hypothalamic CRH mRNA level was found to be approximately the same in the control rats of both strains. In control conditions, the pituitary and plasma ACTH content in ISIAH rats was significantly lower whereas the corticosterone level in the plasma differed from each other in both strain. The restraint stress resulted in a statistically significant increase of the CRH-mRNA in ISIAH rats and not in the WAG rats. Moreover, in spite of the lower ACTH level in stressed ISIAH rats, the corticosterone blood plasma concentration in hypetensive rats was significantly higher. The data obtained confirm the idea that the stress-dependent hypertension might be related to an enhanced sensitivity of the main endocrine links involved in the stress response organization. PMID- 12587272 TI - [Effect of dipeptide vilon on emotional stress resistance in rats]. AB - Effects of synthetic thymomimetic vilon on open field behaviour, immediate early gene c-Fos expression in paraventricular hypothalamus properties of organs sensitive to emotional stress, and characteristics of albumin in the blood plasma in male Wistar rats, were investigated and are discussed in the article. It is shown that intraperitoneal vilon injection rises the resistance against emotional stress according to prognostic indexes open field behaviour. Vilon administration also inhibits hypertrophy of the adrenals, involution of the thymus, and elevates concentration of albumin in the blood plasma. The number of Fos-immunoreactive neurons in the paraventricular hypothalamus was lower after vilon administration especially in rats resistant against emotional stress. PMID- 12587271 TI - [Effect of striatum destruction on the hypothalamo-pituitary-adrenal axis function in adrenalectomized rats]. AB - The influence of the bilateral striatectomy on activation of the hypothalamus pituitary-adrenal axis after unilateral adrenalectomy was investigated. Increasing of the intact adrenal gland's mass in striatectomized rats was less than in the control group. It was combined with decreasing of the production of corticosteroids by intact gland. Such disturbances were more obvious in rats with destruction of the dorsal striatum than the ventral one. These facts suggest the striatum participation, its dorsal zone in particular, in the processes of regulation of the hypothalamus-pituitary-adrenal axis. PMID- 12587273 TI - [Effect of testosterone and progesterone on carboxypeptidase H activity during stress in mice]. AB - It was found that testosterone propionate (3 mg/kg of body weight) and progesterone (1 mg/kg of body weight) partially prevent an augmentation of the carboxypeptidase H activity in the mouse pituitary gland under stress caused by a single intraperitoneal administration of olive oil. In testicles, testosterone prevented augmentation of the enzyme activity within 0.5 hours but increased it through in 4 and 24 hours after treatment. Progesterone in testicles augmented the enzyme activity within 0.5 hours, but not in 4 and 24 hours. Sex steroids were not affected by stressinduced carboxypeptidase H activity in hypothalamus and adrenal medulla. It was concluded that carboxypeptidase H did not take part in integration of hypothalamus-pituitary-adrenal and hypothalamus-pituitary gonadal axis under the stress. PMID- 12587274 TI - [Effect of glutamate on membrane potential and volume of the skeletal muscle fibers in rats following NO-synthase inhibition in vivo]. AB - Cross-sectional area (CSA) of muscle fibers incubated in culture medium 199 for 3 hours dramatically increases, whereas resting membrane potential (RMP) decreases compared to "freshly-isolated" muscles. Both glutamate and sodium nitroprusside prevent these changes. MK-801, a specific inhibitor of NMDA-receptors, eliminates protective effects of glutamate on both CSA and RMP. NO-synthase inhibition in vivo promotes an increase of initial CSA and decrease of mean RMP. Under these conditions, effects of glutamate and sodium nitroprusside on CSA and RMP of denervated muscles are less obvious. It has been concluded that synaptic glutamate is able to participate in regulation of RMP and cell volume in muscle fibers through the activation of postsynaptic NMDA-receptors and muscle NO synthase. PMID- 12587275 TI - [Effect of thyroliberin on membrane potential and pattern of spontaneous activity of neurons of the respiratory center in rats in vitro]. AB - On frontal brainstem slices of rat by means of whole-clamp recordings, we investigated effects of TRH (10(-8) [symbol: see text]) on membrane potential and firing pattern of the neurones in ventrolateral area of the solitary tract nucleus and pre-Botzinger complex. TRH induced a membrane depolarisation and an increase in spontaneous activity of the respiratory centre neurones. After TRH administration, a shortening of time intervals between the beginning of bursts was found in bursting neurones of the pre-Botzinger complex. In some silent neurones, TRH elicited appearance of firing activity, so the silent neurones of the solitary tract nucleus were transformed into tonic while the silent pre Botzinger complex neurones were transformed into bursting ones. Thus, there is a direct regulatory effect of TRH on the respiratory centre neurones at the level of their membrane. PMID- 12587276 TI - [Development of deep hypothermia in rats with limited motor activity]. AB - Changes of the main organism functions (breathing frequency, heart rate and shivering) were investigated under hypothermia in two groups of rats. Animals of the first group were fixed rigidly on the special platform with fixing of head and limbs, and those of the second one--the rats, were placed in a punched cylindrical chamber, inside which they could move freely forward and back. In 2.5 3.0 hours after anaesthesia the rats were placed in a refrigerator (-5 degrees C) until they stop breathing. Cessation of breathing of the first group rats occurred in 1.7 +/- 0.3 hours from the beginning of cooling at body temperature 17.3 +/- 0.6 degrees C and the brain temperature 15.7 +/- 0.5 degrees C. In the second group, a prolonged activation of the frequency of breathing, heart rate and intensity of electrical activity of muscles during 2.5-3.0 hours, was observed. Only in 4.5-5.0 hours, the breathing stopped at rectal temperature 12.3 +/- 1.1 degrees C and the brain temperature 12.9 +/- 0.9 degrees C. In these animals, the time of survival in the cold environment increased considerably and the temperature thresholds of the termination of breathing were lowered. Thus, the activation in the thermo-regulative muscle tone and in shivering muscles provides the most effective resistance against cooling of rats, reducing a surface of heat, dissipation and keeping the temperature of internal areas of body. PMID- 12587277 TI - [Restoration of the respiratory center functions in rats after its complete cold induced paralysis ]. AB - White rats were cooled until cessation of their breathing (about 17.6 degrees C) and then transferred to a 19-20 degrees C room. The breathless rats were administered EDTA (experimental group) and saline (control groups). Within 6 min., warming of the brain stem to 18.6 degrees C caused restoration of breathing in experimental rats whereas the control rats remained breathless. The EDTA binds calcium ions in the cells' cytosol and thus restores the normal regulation of metabolism in the brain cells. PMID- 12587278 TI - [Effect of temperature on impulse activity of cortical neurons in guinea pigs]. AB - The temperature change of incubating fluid from 37 to 24 degrees C results in significant decrease of neuronal spontaneous firing rate in parietal cortex slices. Evoked spike responses to microiontophoretic application of glutamate practically did not change. PMID- 12587279 TI - Clinical pain management techniques for cats. AB - Although pain management is an emerging and popular topic in veterinary medicine, use of analgesics in cats has received little attention relative to their canine counterparts. Some of the difficulty lies in assessment of whether or not a cat is in pain. Simple observation of a cat in a cage relies upon overt expression of pain, and is often inaccurate. Pain scales have been developed that allow a semiquantitative evaluation of the degree of pain an animal may be experiencing. However, treating pain based upon observation of the painful state is less effective than anticipating and preemptively treating pain. This article reviews specific methods for preemptively treating and alleviating pain in the cat. The traditional approach to pain management involves drug administration. Specific categories of agents used in cats include opioids, nonsteroidal anti inflammatories, or alpha-2 agonists. Other modalities of pain management, which are also reviewed, include use of local anesthetic drugs for local and regional analgesia, as well as acupuncture. PMID- 12587280 TI - Onychectomy and its alternatives in the feline patient. AB - Onychectomy and its alternatives are often sought by owners to provide permanent alleviation of scratching behaviors in cats. These alternatives include behavioral modification, nail caps, and deep digital flexor tendonectomy. Although surgical options are commonly performed, they are controversial and have potential complications. Our role as veterinarians is to educate clients to allow them to make the best decision for themselves and their pets. The purpose of this article is to provide an objective review of the positive and negative aspects of onychectomy, describe and discuss alternatives to onychectomy, and review the specific surgical techniques. PMID- 12587281 TI - Nonhealing subcutaneous wounds in the cat and proposed surgical management techniques. AB - Nonhealing wounds in cats are a source of frustration for veterinarians and pet owners alike. These wounds are known for their insidious onset and tendency to recur after standard surgical closure. The presence of a feline wound that fails to heal in the face of appropriate therapy should initiate an extensive diagnostic evaluation for an etiologic agent. Initially animals should be screened for causes of immunosuppression or systemic disease. The local wound environment should then be evaluated through a combination of wound exploration, cytology, histopathology and tissue culture for evidence of a foreign body, neoplasia, or other less common etiologies of these wounds. Many of the less common etiologic agents require specific tissue handling for successful microscopic identification and culture. It is therefore essential for veterinarians to be familiar with the various causes of nonhealing wounds in cats so that laboratories can be alerted of the need for specific diagnostic techniques. Recent surgical and medical advances have the potential to facilitate the successful treatment of these wounds. With improved veterinary awareness of the potential etiologies of nonhealing wounds in cats and improved medical and surgical management techniques many wounds can now successfully be treated. PMID- 12587282 TI - Injection site sarcomas in cats. AB - Feline injection-site sarcomas have mostly been associated with the administration of vaccines. Although the pathogenesis remains largely unknown, it is believed that the inflammatory reaction caused from the injection of the compound leads to uncontrolled proliferation of fibroblasts and myofibroblasts that in a subset of cats undergo malignant transformation. Pretreatment evaluation and proper planning of the therapy are essential. Because of the aggressive nature of these tumors, a multimodality treatment approach is being recommended. A combination of surgery and radiation therapy with or without chemotherapy appears to be providing the best prognosis. PMID- 12587283 TI - Nasopharyngeal polyps in cats. AB - Nasopharyngeal polyps are non-neoplastic, inflammatory growths that arise from the middle ear or the eustachian tube and extend into the pharynx. The exact etiology of nasopharyngeal polyps is unclear; proposed etiologies include a response to chronic upper respiratory tract infection, chronic otitis media, ascending infection from the nasopharynx, or a congenital origin. Clinical signs usually relate to obstruction of the nasopharynx, with Horner's syndrome and head tilt being consistent with otitis media and otitis interna, respectively. Diagnostic tools include digital or visual examination above the soft palate, flexible fiberoptic caudal rhinoscopy, radiography, computed tomography, and magnetic resonance imaging. Ventral bulla osteotomy combined with traction removal of the polyp is the recommended treatment, although traction only followed by prednisolone therapy can be considered in some cases, especially when there is no evidence of otitis media. PMID- 12587285 TI - Screening criteria for feline renal transplant recipients and donors. AB - Renal transplantation is associated with several potential complications. Early selection and appropriate screening of candidates has reduced the morbidity and mortality associated with this procedure in human beings. In veterinary medicine, the important process of identification and screening of candidates for renal transplantation occurs primarily in private general or specialty practice. This article reviews guidelines for screening of candidates for renal transplantation and related publications in both veterinary and human literature in an attempt to provide comprehensive information to practitioners involved in the selection process. It is anticipated that early referral of stable cats for renal transplantation will further improve the likelihood of a successful outcome. PMID- 12587284 TI - Gastrointestinal obstruction. AB - For cats that present with signs of gastrointestinal disease, obstruction is a primary differential. There are numerous reasons of gastrointestinal obstruction in small animals, yet there are several specific causes that are more commonly associated with the cat. These include linear foreign bodies, trichobezoars, focal intestinal neoplasia, feline infectious peritonitis, and megacolon. Clinical signs related to gastrointestinal obstruction consist of vomiting, diarrhea, constipation, tenesmus, anorexia, or weight loss. The course and onset of disease depends on the rate at which the obstruction develops and whether the obstruction is partial and complete. The diagnosis of obstruction is typically suspected based on clinical presentation and palpation of an abdominal mass. Diagnostics tools are used for definite diagnosis and determination of location within the gastrointestinal tract. Surgical treatment is dependent on the etiology of the obstruction and various techniques are employed to remove the obstruction and prevent recurrence. PMID- 12587286 TI - Feline orthopedics. AB - Although many aspects of canine care do cross over to the feline patient, the adage "cats are not small dogs" holds more true today than ever. The area of feline orthopedics is no different. The purpose of this article is to familiarize the reader with the more common feline orthopedic problems. Although many nonsurgical aspects are covered, the majority of text discusses surgical orthopedic problems. In addition, situations considered unique to the feline patient are addressed, including preoperative, perioperative, and postoperative concerns. Diagnostics and implant systems are also covered. PMID- 12587287 TI - Rapid communication: partitioning of persistent lipophilic compounds, including dioxins, between human milk lipid and blood lipid: an initial assessment. AB - A systematic program of sampling and analysis of blood serum for dioxins, furans, and dioxinlike polychlorinated biphenyls (PCBs) has been initiated in the United States through the National Health and Nutrition Examination Survey (NHANES) program. While such data could potentially be used to estimate population-level changes in human milk lipid concentrations of chemicals, such estimates would depend on understanding the relationship between human blood lipid and milk lipid concentrations of the compounds of interest. For dioxins and furans, extremely limited data in humans currently exist for paired blood/milk samples. These data reviewed in this article, support the hypothesis that, over a population and across time, human milk lipid levels of these compounds generally reflect blood lipid levels. However, these data also suggest that significant variations in these ratios are possible among individuals and at various times. PMID- 12587288 TI - Inflammatory effects of inhaled endotoxin-contaminated metal working fluid aerosols in rats. AB - Exposure to aerosols generated from water-soluble metal-working fluids (MWF) is associated with numerous respiratory symptoms consistent with an acute pulmonary inflammatory event. Previous studies in mice and guinea pigs have implicated endotoxin contamination of MWF as the causative agent responsible for inducing pulmonary neutrophilia and decrements in airway conductance. However, little information is known about the relationship between endotoxin-contaminated MWF exposure and changes in airway physiology. The present study, utilizing a rat model, has demonstrated that exposure to 10 mg/m3 with endotoxin (0 to 3.2 micrograms/m3) resulted in a time- and concentration-dependent migration of neutrophils in the lung tissue's interstitial spaces as well as the lavageable airways. In contrast to other airborne toxicants, where neutrophil infiltration of the lung has been associated with hyperresponsive airways, the endotoxin induced neutrophilia observed in the present study was not associated with airway hyperresponsiveness to challenge with the muscarinic agent methacholine or with permeability damage to the lung. Bronchoalveolar lavage (BAL)-recovered neutrophils demonstrated no adverse effects as a result of endotoxin-contaminated MWF exposure. In contrast, a population of alveolar macrophages was observed to be enlarged in size and demonstrated an increased sensitivity to oxidative metabolism when challenged with phorbol myristate acetate, consistent with being at a relatively high state of activation. These results suggest that while endotoxin contamination of MWF is capable of producing an acute inflammatory event, other predisposition factors may be required to induce alterations in pulmonary physiology. PMID- 12587289 TI - Exposure to particulate 1-->3-beta-glucans induces greater pulmonary toxicity than soluble 1-->3-beta-glucans in rats. AB - 1-->3-beta-Glucans, derived from the inner cell wall of yeasts and fungi, are commonly found in indoor air dust samples and have been implicated in organic dust toxic syndrome. In a previous study, it was reported that 1-->3-beta-glucan (zymosan A) induced acute pulmonary inflammation in rats. This study investigates which form of 1-->3-beta-glucans, particulate or soluble, is more potent in inducing pulmonary inflammation. Zymosan A was suspended in 0.25 N NaOH for 30 min, neutralized, dialyzed for 2 d using deionized water, and particulate and soluble fractions were collected. Male Sprague-Dawley rats were exposed via intratracheal instillation to NaOH-soluble or NaOH-insoluble zymosan A. At 18 h postexposure, various indicators of pulmonary response were monitored, including indicators of lung damage, such as serum albumin concentration and lactate dehydrogenase (LDH) activity in acellular bronchoalveolar lavage fluid. Inflammation was characterized by an increase in lavageable polymorphonuclear leukocytes (PMN). Pulmonary irritation (breathing frequency increase) and oxidant production (nitric oxide and chemiluminescence, CL) were also monitored. Exposure to the particulate form of NaOH-treated zymosan produced a significant increase in all these indicators. In contrast, rats exposed to the NaOH-soluble fraction were not markedly affected except for LDH, PMN, and CL. However, these increases were significantly less than with exposure to NaOH-insoluble zymosan. Therefore, results demonstrate that particulate zymosan A is more potent in inducing pulmonary inflammation and damage in rats than the soluble form of this beta glucan. PMID- 12587290 TI - Impact of coexposure to ozone on the carcinogenic potential of inhaled chromium. 1. effects on retention and on extra- and intracellular distribution. AB - A health hazard to welders is development of lung cancer. It is believed that this is likely due, in part, to the presence in welding fumes of several hexavalent chromium (Cr[VI]) species, whose solubility depends primarily on which process (i.e., manual metal arc verus metal-inert gas) is used. However, inhalation of Cr alone is uncommon in this setting. Thus, an examination of potential contributions from other coinhalants in creating or enhancing conditions whereby inhaled fume-associated Cr (primarily the insoluble forms) may initiate cancer is critical to increasing our understanding and preventing this particular occupational disease. One major chemical species formed and released during welding is ozone (O3). Though implications of adverse pulmonary effects from individual exposure to Cr or O3 have been investigated, those from simultaneous exposure are unclear. To begin to address whether the carcinogenic potential of insoluble Cr[VI] agents might be enhanced in hosts inhaling mixtures of Cr and O3 versus Cr alone, analyses of total lung Cr burden, Cr retention in lung epithelium and interstitium, and potential shifts in lung cell distribution of Cr from the cytoplasm to nuclei were undertaken in F-344 rats exposed nose only (5 h/d, 5 d/wk for up to 48 wk) to an extrapolated occupationally relevant level of Cr (360 micrograms Cr/m3 as calcium chromate) alone and in combination with 0.3 ppm O3. Overall, there was only a nominal effect from O3 on Cr retention or on distribution of Cr particles among extracellular sites and within lung cells. However, there were O3-related effects upon mechanisms for clearing the Cr from the deep lung, specifically at the levels of particle uptake and postphagocytic/endocytic processing by macrophages. This O3 exposure-related shift in normal pulmonary clearance might potentially increase the health risk in workers exposed to other insoluble or poorly soluble carcinogenic Cr compounds. PMID- 12587291 TI - Testicular germ-cell apoptosis in stressed rats following combined exposure to pyridostigmine bromide, N,N-diethyl m-toluamide (DEET), and permethrin. AB - This study reports and characterizes the testicular apoptosis following daily exposure of male Sprague-Dawley rats to subchronic combined doses of pyridostigmine bromide (PB, 1.3 mg/kg/d in water, oral), a drug used for treatment of myasthenia gravis and prophylactic treatment against nerve agents during the Persian Gulf War; the insect repellent N,N-diethyl m-toluamide (DEET, 40 mg/kg/d in ethanol, dermal); and the insecticide permethrin (0.13 mg/kg in ethanol, dermal), with and without stress for 28 d. Combined exposure to these chemicals was implicated in the development of illnesses including genitourinary disorders among many veterans of the Persian Gulf War. Previous studies from this laboratory have shown that exposure to combination of these chemicals produced greater toxicity compared to single components. Exposure to stress alone did not cause any significant histopathological alterations in the testes. Administration of combination of these chemicals induced apoptosis in rat testicular germ cells, Sertoli cells, and Leydig cells, as well as in the endothelial lining of the blood vessels. Testicular damage was significantly augmented when the animals were further exposed to a combination of chemicals and stress. Histopathological examination of testicular tissue sections showed that apoptosis was confined to the basal germ cells and spermatocytes, indicating suppression of spermatogenesis. Increased apoptosis of testicular cells coincided, in timing and localization, with increased expression of the apoptosis-promoting proteins Bax and p53. Furthermore, significant increase of 3-nitrotyrosine immunostaining in the testis revealed oxidative and/or nitrosation induction of cell death. In conclusion, combined exposure to real-life doses of test compounds caused germ cell apoptosis that was significantly enhanced by stress. PMID- 12587292 TI - Physical effects of chlorpyrifos on two stages of xenopus laevis. AB - The effects of a 96-h static exposure to chlorpyrifos were examined in two developmental stages of Xenopus laevis (premetamorph and metamorph). Measures of effect included body length, mass, and swimming ability. Most parameters indicate that metamorphs were more sensitive than premetamorphs. For larvae exposed as premetamorphs, body length and mass were significantly lower after exposure to 0.001 mg/L and 0.1 mg/L, respectively. Premetamorph swimming activity was increased by a concentration of 0.001 mg/L. Body length and swimming activity were the most sensitive endpoints in metamorphs, showing effects at 0.0001 mg/L. The corresponding body mass was not affected until exposure to 0.1 mg/L. Based on recent environmental data, chlorpyrifos has a 0.1-32.8% probability of exceeding a 96-h time-weighted average of 0.0001 mg/L in U.S. surface waters. PMID- 12587294 TI - [Characteristics of modern diagnosis of diseases caused by Chlamydia bacteria]. AB - Chlamidia psitacci, Chlamidia trachomatis, Chlamidia pneumoniae are intracellular bacterial parasites causing ornithosis, trachoma, conjuctivitis, urogenital, respiratory infections, arthritis, Reiter's disease. The role is discussed of the above microorganisms in the development of cardiological, neurological disorders. The past years have seen further progress in microbiological diagnosis but information now available on, in the first place, Chlamidia pneum., is scarce. In most clinical studies they use a serological diagnosis which is not without faults, with the cultural method and polymerization chain reaction being most informative diagnostic tools. The diagnostic value of the above-mentioned reaction will, we believe, increase with its improvement. In the preliminary diagnosis, cytomorphological methods can be made use of. PMID- 12587293 TI - Malondialdehyde-protein adducts in the spleens of aniline-treated rats: immunochemical detection and localization. AB - Previously it was reported that aniline exposure in rats induces increased lipid peroxidation and formation of malondialdehyde (MDA)-protein adducts in the spleen. In order to further elucidate the role of MDA-protein adducts in the splenic toxicity of aniline, studies were conducted to detect and localize these adducts in the spleen. Rabbit polyclonal antisera to MDA-keyhole limpet hemocyanin were employed for immunohistochemical localization and Western blot analyses of MDA-protein adducts in the spleens of aniline-treated (65 mg/kg/d aniline in the drinking water for 30 d) and control rats. For immunohistochemical localization of MDA-protein adducts in the spleen, a new approach using alkaline phosphatase-fast red (red color) to demonstrate bound primary antibodies was adopted, providing a sharper and increased contrast compared to horseradish peroxidase-diaminobenzidine (brown color) methodology. This new approach allowed us to differentiate the changes in aniline-treated spleens, which had extensive brownish deposits of iron proteins. Spleens from aniline-treated rats showed intense staining for these adducts in the red pulp areas (where iron was also localized), especially within the sinusoidal macrophages. Spleens from control rats showed only mild staining for adducts and only traces of iron. Western blot analyses of splenic microsomal proteins from aniline-treated and control rats showed the presence of 13 different MDA-modified proteins. However, 26-, 32-, and 14-kD proteins were more prominent in the aniline-treated rats. The colocalization of MDA-protein adducts with iron in the red pulp of the spleen suggests that iron-catalyzed lipid peroxidation leading to formation of MDA protein adducts could be a potential mechanism for splenic toxicity of aniline. PMID- 12587295 TI - [Approaches to pathogenetic immunotherapy of HIV-infections]. AB - Substantiated in the paper is urgency of development of pathogenetically adequate immunotherapeutic approaches to the treatment of HIV-infection, that is supposed to have good prospects. Consideration is given to experimental and clinical-and immunological findings concerning effects of different immunomoderivative drug preparations and of their synthetic analogues on the trigger mechanisms, with sequelae of their development in the cytokin-chemokin system outlined. A possibility is shown based on principle of our using with an immunotherapeutic purpose some cytokins, soluble receptors, antibodies to cytokins and receptors, those antibodies and ligands combined with toxins, chemical compounds affecting the synthesis of cytokin and other biologically active substances as well. Unspecific active immunotherapy is regarded as a good supplement to antiretroviral therapy preventing development of resistance to drugs. PMID- 12587296 TI - [Antibiotics in the treatment of syphilis (history, modern state of the problem, perspectives)]. AB - Submitted in the report are data in brief outline on therapy of syphylis with mercury, arsenic, bismuth preparations and their use-associated complications. Historical stages of discovery, production, use of penicillin and its durant forms are described together with those of other antibiotics in the treatment os syphylitic infection in the USSR and abroad. A comparative efficiency of antibioticotherapy versus therapeutic modalities (combined and permanent) is given. PMID- 12587297 TI - [Diagnostics and prediction of development of pulmonary complications in acute myocardial infarction (morphological validation of the use of non-invasive investigational methods]. AB - With the aid of gaschromatographic techniques a fatty-acid composition was studied of lipids of the expired air condensate and sweat of patients in uncomplicated/complicated course of acute myocardial infarction versus morphological changes in the blood-air barrier. The complicated course of the condition vs uncomplicated one has been found out to be accompanied by a significant increase in the lipid content of the polyunsaturated fatty acids- linoleic and arachidonic acids--in the studied biological objects, which fact can be used in diagnosis and prognostication of development of complications in the acute period of myocardial infarction. PMID- 12587298 TI - [Morphological changes in the gastric mucosa in patients with peptic ulcer of different localisation]. AB - Morphological changes were studied of gastritis in patients with ulcer localized in the upper, middle thirds of the stomach, in its antral segment, and in patients with ulcer in the bulb of duodenum, the former two groups of patients displaying mild inflammation/activity if any, the latter ones showing a high to moderate degree of inflammation and activity of gastritis identifiable in the antral segment of the stomach. The findings secured can be helpful in the diagnosis of gastritis of different localization, prognostication of the course of the illness, and adoption of adequate therapy. PMID- 12587299 TI - [Spontaneous and Fas-induced apoptosis of mononuclear leukocytes in rheumatoid arthritis]. AB - A study was made into the ability of mononuclear leukocytes recovered from blood of 77 patients with rheumatiod arthritis (RA), 17 patients with osteoarthrosis deformans (OAD), 18 patients with Chlamydia-associated reactive arthritis (ReA), and 20 healthy controls, to precipitate spontaneous and Fas-induced apoptosis. RA patients versus healthy persons and patients with OAD and ReA displayed lower parameters characterizing spontaneous and Fas-induced apoptosis. A correlation was established between the ability of T- and B-lymphocytes to usher in Fas induced apoptosis during incubation with RhHA and KonA and degree of RA activity. PMID- 12587300 TI - [Methodics and aspects of postmortem forensic psychiatric expertise in a civil law suit]. AB - With the purpose of studying issues concerning conducting a postmortem forensic psychiatric examination (FPE) in a lawsuit, getting an expert opinion, an analysis was performed of 55 cases of postmortem FPE with respect to those subjects with mental disturbances secondary to vascular affection of the brain (cerebral atherosclerosis and relevant complications), having been within the competence of experts. Measures are submitted designed to do away with the disadvantages spotted. PMID- 12587301 TI - [Phenomenon of religious experience in the structure of feelings of individuals with mental disturbances]. AB - Topicality is substantiated of development and use of psychomatric methods of investigation into the phenomena of religious experience in a diagnostic and expert psychiatric practice. Worthwhile lines of using such methods are outlined. The author's data are submitted on adaptation of the Religiosity Scale to the Ukrainean population and on studying phenomena revelant to the bioenergoinformation healing practice. PMID- 12587302 TI - [Pathomorphological changes in the uterine wall in gestosis]. AB - With the purpose of studying a morphological substrate of the uterine contractile function disorder in gestosis, an investigation was conducted designed to study biopsy specimens of the uterine wall having been taken during the cesarean section in 80 parturient women. The womb of those parturient women having mild and severe gestosis complicated by uterine inertia (40 observations) or premature dethachement of the normally situated placenta (8 cases) was studied together with that of controls (32 cases). The indentified micro-circulatory disturbances and alterative changes in the myometrium structures in gestosis are regarded as one of the generalized blood circulation disorders, hemostasis (dissaminated intravascular blood coagulation) and systemic abnormalities of metabolism presenting as a multiorgan failure developing in the pathology under consideration. Microcirculatory disturbances and hypoxia-related dystrophic changes in the smooth myoblasts and connective tissue leading to its fibrosis- all these constitute a morphological substrate of changes in the contractile function of the womb. PMID- 12587304 TI - [Clinical and hematological aspects of follicular lymphoma]. AB - A comprehensive clinical-and-laboratory examination of patients with non Hodgkin's malignant lymphomas revealed a lack of correspondence between results of histological and immunophenotypical investigations. The most characteristic feature appeared to be the presence of CD10, CD11a, and CD35 antigens. Absence of CD95 antigen on tumour cells is regarded as an unfavourable prognostic sign in patients with follicular lymphomas as is presence of CD10 antigen on the above cells. PMID- 12587303 TI - [Clinical and immunological studies of the therapeutical effect of cytokines combined with 5-fluorouracil in metastatic kidney cancer]. AB - The authors present the results of a six-year clinicoimmunological study of a therapeutic effect of cytkins combined with 5-fluoruracyl in metastatic renal cell carcinoma. Two therapeutic regiments have been used: rhIFN-a + 5-FU and rhIL 2 + RHifn-A + 5-FU. In cytokin-sensitive patients, both therapy protocols vrs conventional therapeutic alternatives (cytostatics, hormones, irradiation) have been shown to increase the frequency of achievement of remission by objective scoring and life span of the patients. There was an improvement in patients on having received the complex with IL-2 but a higher therapeutic effect appeared to be accompanied by substantial side effects. Recommendatory measures well-targeted to those patients with metastatic renal-cell carcinoma who are to be placed on cytokinotherapy are presented together with immunological indices to monitor the treatments administered and prognosis. PMID- 12587305 TI - [Ultrasonography control of seroma development in patients after radical breast surgery]. AB - Axillary lymphadenectomy, irrespective of volume of surgical intervention, leads to formation of a cavity in the axillary region, its sizes affecting the frequency of postoperative seroma and other complications. Sixty-nine patients with breast cancer underwent ultrasonography for the operative wound, with the latter drained (n-25)/no drain (n = 44). Use of ultrasound for the operative wound permits monitoring the process of seroma formation, carrying out an adequate aspiration of the liquid, avoiding wound draining. PMID- 12587306 TI - [Sparing therapy of aged patients with cancer of the endometrium, uterine cervix, and vulva]. AB - Outlined in the article are main principles of organ-preserving and sparing treatment of aged female patients with carcinoma of the body of the womb and vulva. Indications are determined for surgical management of the above medical problem in elderly women. Revelant treatment options are described. PMID- 12587307 TI - [Ways for improvement of medical, occupational, and social rehabilitation of patients operated for complicated gastroduodenal ulcer]. AB - Based on the analysis of the early post-operative period in 1588 patients and long-term results in 667 patients having been operated for complicated gastroduodenal ulcers, algorithms are submitted of medical, occupational, and social rehabilitation, which fact permitted improving results of surgical treatment. The proposed rehabilitation project envisages staging and continuity of measures aimed to restore the patients' capacity for work and their social status. PMID- 12587308 TI - [Chlamydiosis of the upper sections of the digestive tract]. AB - The experience is submitted gained with the diagnosis and treatment of Chlamydia trachomatis-associated esophagitis. Used in the study were laboratory and instrumental methods of diagnosis, with their merits and demerits outlined. It is pointed out that complex diagnosis of Chlamydia trachomatis-produced esophagitis is important, the treatment involving institution of anti-bacterial and immunomodulating therapies. PMID- 12587309 TI - [The functional status of some endocrine glands in children with enzymopathy of the small intestine]. AB - As many as 53 children with enzymopathies of the small intestine aged 9 months-14 years were examined. Of these, 25 had celiac disease, 27 presenting with Gee Herter syndrome. In all patients, there have been studied patterns of hydrocortisone, thyroid hormones, thyrotrophin, insulin. Ascertained in the above pathology was a decline in the functional activity of the hypophysis, thyroid gland, adrenals, insular apparatus. The degree of the hormonal inadequcy has been found to be related to the form and severity of enzymopathy of the small intestine. The lowest indices for T3, T4, TTH, hydrocortisone, insulin appeared to be in Gee-Herter syndrome. PMID- 12587310 TI - [Biochemical investigation of patients with jaundice of tumor genesis]. AB - The study comprised surgical patients presenting with tumours of the extrahepatic biliferous ducts. The above patients revealed a high activity of gamma glutamyltransferase, alkaline phosphatase at the expense of the hepatic isoenzye, to a lesser extent--of asparaginaminotransferase, alaninaminotransferase, a high level of conjugated bilirubin, an insignificant increase in the level of cholesterol and triglycerides. The above biochemical indices combined with results of other methods of investigation can be used for early diagnosis of mechanical jaundice of the tumour genesis. PMID- 12587312 TI - [Biocontrollable electrostimulation in clinics of nervous diseases]. AB - In the article, mechanisms are described of action of biocontrollable transcutaneous electrostimulation with making use of the device "Skenar"; indications and contraindications for the suggested method to be employed in revelant clinical settings are given. PMID- 12587311 TI - [Long-term results of treatment of tuberculosis in patients who had not received a full basic course of antimycobacterial therapy]. AB - In those TB patients who had not received a full basic course of antimycobacterial therapy, long-term results of treatment are much worse (development of early and late recurrences comes to be more commonly seen--by as high as 6.43 and 5.03% respectively; transition to a chronic form by 20.29%, case mortality by 6.04%, with clinical cure recordable by 37.75% more seldom) than in those patients having received a full basic course of the above therapy. PMID- 12587313 TI - [Clinical and dynamic features of pathological craving for narcotics in patients with opioid (heroin) addiction]. AB - Particular features have been studied of the clinical picture and time-related course of an abnormal urge to consume narcotics in patients with opiumism. The time-related characteristics of the pathological urge undergo qualitative changes with progression of the illness from the discrete character of actualization to a continuous maintenance of the addiction in the active state. Variants of the pathological urge to opiates dynamics, characteristic of different stages of opium addiction are highlighted. PMID- 12587314 TI - [Hormonomodulating effect of the mineral water Naftusia as a basis of efficiency of sanatorium treatment of patients with diabetes mellitus]. AB - Effects were studied of a course intake of the mineral water Naftusya on insulin secretion in patients with latent and mild forms of diabetes mellitus. The gastroenteropancreatic endocrine system takes a direct part in formation of the integral bodily reaction to the mineral water intake. The possibility of our using drinking mineral waters to correct functions of the above system is of a great practical interest in aspects of health-resort therapy of diabetes mellitus and other medical problems. PMID- 12587315 TI - [Clinical and epidemiological characteristics of marseilles (mediterranean tick borne) fever in the Crimea Autonomous Republic]. AB - In the paper, new data are submitted on natural focality of Marseilles fever in the territory of the Crimean Peninsula. Identified in the above territory was a high activity of previously unknown natural foci that manifested themselves by an epidemic outbreak (Saki, 1996) and by sporadic diseases in people. A clinical-and epidemiological characterization is given of rickettsiosis in the Crimea Autonomous Republic. PMID- 12587316 TI - [Dynamics of clinical and biochemical indices in patients with primary liver cancer during endovascular chemotherapy]. AB - Exposed to endovascular chemotherapy were twenty patients with inoperable primary carcinoma of the liver. There was an instant rise in the posttreatment versus basal clinical-and-biochemical indices and their direct correspondence with the functional stage of the primary disease in the liver. The improvement in severity of the condition was more poor in stage III patients. The further the progression of the functional stage, the higher were efficacies of the therapies applied. The author considers it expediant that in functional stage II and III patients, chemotherapy be combined with detoxication therapy. Early correction of the functional condition of the liver will, we believe, help in improving results of the treatments administered. PMID- 12587317 TI - [Thoracoscopy in verification of thoracal endometriosis]. AB - The authors submit data from the published literature, showing that endometriosis is a grave disease in young women of reproductive age. The incidence of endometriosis tends to be on the increase. At the same time, cases have come to be seen of polyorgan localization of the process, the thoracal one included. The paper contains a presentation of a case of thoracal endometriosis accompanied by spontaneous pneumothorax diagnosed after her having been ill with the disease for ten years. Arguments are submitted in favour of a pleural investigation to be conducted in patients with spontaneous pneumothorax, which reasoning determines further therapeutic policy in such patients. In the authors' opinion, it is thoracotomy that is to be embarked upon, the volume of intervention being determined by revision of organs of the pleural cavity. PMID- 12587318 TI - [Morphofunctional aspects of cardioprotective effect of propranolol in modelling of chronic ischemic heart disease]. AB - In experiments involving simulation of ischemic heart disease in adult rabbits, morphofunctional aspects were studied of cardioprotective action of propranolol on the contractile myocardium. With the aid of histological and histochemical, electron microscopic tests, the drug under consideration has been shown to be endowed with membrane-stabilizing properties, quite aside its receptor-mediated effects on Ca2+ metabolism known for. A decrease in its excessive ingress into the sarcoplasma comes, in its turn, to improve energy and plastic maintenance of the myocardial functions, to moderate a pathological reorganization and destructive changes in cardiomyocytes, thus promoting the contractile myocardium resistance to recurrent coronary crises. PMID- 12587319 TI - [Changes in some parameters of nitric oxide synthesis in the early periods of development of hepatocarcinogenesis]. AB - Our objective in this study was to determine some indices for the synthesis of nitric oxide in the early periods of diethylnitrosoamine-induced (10 mg/kg 5 times a week for two months) hepatocarcinogenesis in mature male rats (n = 100) and to gain insights into their relation to lipid peroxidation process. During the time-related course of hepatocarcinogenesis the blood serum level of L arginin has been found to change significantly with time of the study while that of malonic dialdehyde tended to be on the increase. The SOD activity increased somewhat during the period of initiation, with its significant reduction recordable during the period of promotion and a striking increase in the initial period of progression. Unlike SOD, catalase was showing a tendency to increase after some decline in its activity. PMID- 12587320 TI - [Structure in air-blood barrier in experimental pneumonia and chronic alcohol intoxication]. AB - Ultrastructural changes were studied in components of the blood-air barrier in pneumonia in the presence of chronic alcoholic intoxication under exposure to a substitution surfactant therapy. The data obtained suggest to us that use of a replacement surfactant therapy promotes reduction of edematous events and dystrophic-and-destructive changes in cells of the parenchyma of the lungs increasing the functional activity of alveolar macrophages and type II alveolocytes. PMID- 12587322 TI - [The study and comparative evaluation of the actoprotective activity of ATF-LONG in experiment]. AB - In the paper, results are reflected of a swimming test in 78 laboratory rats. ATF LONG has been shown to be capable of augmenting the animals' powers of endurance during loading tests. The established dose-dependent actoprotective activity of ATF-LONG is superior to that of adenosine triphosphate, trimetasidine (preductal), and mildronate. Possible pharmacological mechanisms of its actoprotective activity are discussed. The authors come to the conclusion that performance capability gets improved under exposure to purine nucleotides. PMID- 12587321 TI - [Lysosomal enzymes of the liver and kidney in unspecific purulent kidney disease]. AB - Unspecific inflammatory lesions of the urinary tract are known to be a vital question in urology and nephrology of our country and in foreign ones. Used in experiments were rats of mixed population. Lysosomal enzymes were studied in homogenates of the liver and contralateral kidney tissues and in the blood serum as well 1, 3, 6 days after simulation of the pathology experimental model. Activities were measured of such lysosomal enzymes as acid DNA-ase and RNA-ase, cathepsin, and acid phosphatase. Results of the studies made showed that in unspecific purulent affection of the kidneys there occurs an activation of acid blood hydrolases followed by their activation in the liver. The contralateral kidney, because of it being linked in functioning with the liver, remains relatively "inert" with respect to activation of lysosomal enzymes. PMID- 12587323 TI - [Stability of neurohumoral effects of losartan combined with metoprolol in chronic cardiac low output]. AB - Combined therapy with lozartan and metoprolol in patients with chronic cardiac insufficiency has a consistent effect on the levels of epinephrine, angoitensin II, vasopressin, atrial naturiuretic factor, insulin, endothelline, thromboxane B2, guanosine monophosphate, 6-keto prostaglandin F1 alpha, and bradikinin. As to norepinephrine, there has been noted an inconsistent therapeutic effect. It is suggested that the time-related course of a long-term therapy might be associated with a formation of the aldosterone "escape" phenomenon. PMID- 12587325 TI - [Propofol--new highly effective intravenous anesthetic]. AB - A review is presented of revelant medical literature concerning potentialities of a new intravenous anaesthetic propofol that has come into widespread use as a means of inducing and maintaining anaesthesia, short-term and long-term sedation. Propofol allows an extremely prompt induction of anaesthesia and postanaesthetic recovery to be achieved, with the depth of anaesthesia easily controlled. PMID- 12587324 TI - [Effect of enape combined with nebilet in hypertension]. AB - Patients with stage II hypertensive disease have been shown to derive apparent benefit from treatment with a combination of the beta-adrenoblocker nebivolol and inhibitor of the angiotensin-converting enzyme enap that was evidenced by improvement in hemodynamics, in echocardiographic characteristics promoting a decline in the activity of the basic enzyme of the tissue formation angiotensin II chimase and an increase in the level of its most important inhibitor alpha-2 macroglobulin. PMID- 12587327 TI - [Microbiological validation of the composition of "decaceol", an antiseptic prolonged-release drug]. AB - With the aid of different microbiological methods, antimicrobal properties of decametoxine, a home-produced antiseptic, were studied together with those of the synthetic zoelite NaA-base drug preparation. The antimicrobial activity of the above drug has been shown to be of a synergistic character that makes it a very promising medical agent which will, we believe, come to be widely used in medical practice. PMID- 12587328 TI - [Clinical stages of lead intoxication]. AB - The authors, on performing a critical analysis of reported data on clinical manifestations of lead intoxication, suggest that the initial manifestations thereof presenting as changes in no other indices but laboratory ones, be regarded as a preclinical stage. Aside from generally recognized concepts of "mild" and "prominent" stages in lead intoxication the authors consider it logical and expediant to introduce a moderately severe stage manifested by vegetosensory polyneuropathy, astheno-vegetative syndrome, dyskinesia of the biliary and intestinal tracts, spastic colitis. It is in this very stage of lead intoxication that the patient, to the author's thinking, is to be assigned to some other job where he/she will not be exposed to lead or other adverse occupational factors. This will, we believe, permit preventing development of a clinically significant stage of lead intoxication manifested by encephalopathy, motor form of polyneuropathy, lead colic, anemia and hepatitis. PMID- 12587326 TI - [Immunologic monitoring in transplantation of hydrophilic polyacrylamide gel]. AB - There was no significant difference in the level of basic immunological indices between those female patients having received a breast prothesis with making use of the polyacrilamide gel (PAAG) interfall and controls, subjects who had not received a breast prothesis, and group of patients in whom the gel was administered by injection, which fact corraborates the conclusion about immunological tolerance of the gel, with no dependence on the way in which it is used being noted. PMID- 12587329 TI - [Substantial experience in organisation of around-the-clock angioplasty emergency care in regional clinical hospital]. AB - The submitted indices by separate nozological entities over the period 1996-2000 permit representing patterns of acute disorders and injuries of the great vessels in 779 patients, which fact can be used as a criterion for assessment of quality of work of the angiosurgeon making emergency calls on sick persons in the above category. It will also, we believe, be helpful in developing standards for examination and treatment in order that emergency surgery might be delivered to such patients. PMID- 12587330 TI - [Prevalence, morbidity, and mortality in cerebrovascular diseases in Mariupol]. AB - A comparative analysis was done of prevalence, incidence rates, and mortality from cerebrovascular disorders (CVD), including cerebral insults, in Mariupol over the period 1999-2000. Submitted in the paper are CVD epidemiological findings from the Donetsk region, Ukraine. Of much interest to us was studying prevalence, risk factors for CVD development among those workers occupied in labour at large industrial enterprises in Mariupol, with which purpose sampling population studies were made among workers occupied in labour at the Illyich Integrated Iron-and-Steel Works and seaport. During the course of the above studies epidemiological indices were examined together with risk factors for development of cerebrovascular disorders. PMID- 12587331 TI - [The role of a family doctor in oncological care of the population]. AB - The actual tasks faced by the family physician in the field of oncology can be considered in eight lines of activities according to successive stages of management of patients, that include identification and monitoring of risk factors; screening of early forms of cancer; diagnosis that must be based on results of the screening during the regular health checkups of patients and their applying for medical advice; primary finding out the extent of the process (staging); rendering the patient and his/her family phychological support; the family physician's work together with that of the oncologist; keeping the patient and his/her family under medical surveillance; terminal stages management. PMID- 12587332 TI - [Apollinarii Grigor'evich Podrez, professor of surgery in the Khar'kov university ( on 150th anniversary of the birthday)]. AB - The article is devoted to an outstanding surgeon of the late XIXth century Professor A. G. Podrez. His scientific work is described together with his social activities. PMID- 12587333 TI - [Domestic violence. Epidemiologic data]. AB - Domestic violence has been little studied in France. However a national survey (Enquete Nationale sur les Violences Envers les Femmes en France, ENVEFF) shows that 9% of the women had experienced a domestic violence, including 2.3% very severe. Even if these violences are related to several characteristics or situation of the couples, they are observed in every social or cultural group. Health professionals have a role to play in the identification of the problem and the related care. PMID- 12587334 TI - [Consequences of domestic violence on women's health and their management in primary health care]. AB - Because of its frequency and impact on women's health, domestic violence is an important public health issue. Psychological and psychosomatic disorders, pathological pregnancies as well as traumatic and gynaecological conditions are the main consequences of such violence, which may arise at a distant term after the events that constitute it. Health professionals, particularly general practitioners, emergency doctors, gynaecologists, obstetricians and midwives are on the frontline in identifying the victims and providing care to them. Therefore, there is a need to make a wide range of specific and relevant information readily available to these professionals and a call to renew their practices and increase partnerships. PMID- 12587335 TI - [Consequences of family violence on children's health]. AB - Family violence is frequent. It has significant repercussions upon the child's health whatever his age: functional troubles, psychosocial dwarfism, behavior troubles, depressive condition, teenagers' suicide attempts. PMID- 12587336 TI - [Child abuse]. AB - Child abuse and neglect a very difficult subject for physician as for every body. How can we help them? The part of hospital and reference hospital team is essential. PMID- 12587337 TI - [Incest today]. AB - The issue explored is incest against children. This paper summarizes the changes on knowledge on this topic since the years 1850 to nowadays. The main focus is placed on the specificity of incest and sexual aggression towards minors as criminal behaviours. Finally, the lake of research, which leaves many questions unanswered, is underlined. PMID- 12587338 TI - [Cruel and violent children]. AB - Three to 4% of parents are regularly battered by their children, at the adolescence. Before the appears of violence, there is an escalation of pathologic interaction between parents and child. Comorbidity is usual: problem behavior, impairments in school functionning, antisocial disorders, etc.... Sometimes violence seems to appear at the adolescence in families without any problem. Very often there are many family conflicts and discrepancy between parents about education of their children. Prevention is the better treatment with an early intervention as soon as possible. PMID- 12587339 TI - [Heart failure with preserved left ventricular function: clinical, echocardiographic, and clinical course features. Prognostic factors]. AB - Heart failure is clinically associated with inadequate myocardial contraction, a significant reduction of left ventricular systolic function and ejection fraction and a cardiac enlargement. Some studies have reported that patients with symptomatic heart failure may have an impaired left ventricular filling with a normal or preserved left ventricular systolic function and an ejection fraction > 45%. These patients have a "diastolic heart failure" often neglected or misdiagnosed. The aims of our study is to describe clinical, echocardiographic and hemodynamic characteristics of 64 patients hospitalized for symptomatic heart failure, to determine possible variables with prognosis relevance, and for evaluating the severity of this diastolic left ventricular dysfunction. All patients were assessed by physical and radiographic examination, 12 leads electrocardiogram, and usual laboratory tests. The internal diameter of left atrium and left ventricular end diastolic and tele-systolic diameter were measured following the recommendations of the American Society of Echocardiography, Ejection fraction was determined following Simpson's method. Left ventricular filling patterns were evaluated by pulsed Doppler mitral or venous pulmonary flow. The following parameters were assessed: maximum velocity of E and A waves, E/A ratio, E wave deceleration time and isovolumic relaxation time. The patients were studied following Appleton's classification. 45 patients were submitted to left heart catheterization and coronary angiography. All subjects were routinely followed by cardiologic examinations and the mean follow up is 18 +/- 4, 5 months. 29 women (45.3%) and 35 men with a mean age of 72.5 +/- 3.2 years were included in this study. Left ventricular ejection fraction was in mean 48.5 +/- 4.2%. 65% of patients had ischemic cardiomyopathy with severe coronary stenosis > 50%, often associated with hypertension. 52% of patients had hypertensive heart disease and 38% were diabetics. 34 patients were re hospitalized for recurrent heart failure despite medical treatment with diuretics, ACE inhibitors (90% of patients), beta-blockers, (37%) or nitrates (36%). 24 patients have been treated by coronary angioplasty. In hospital mortality was 6.2% and during the follow up at 18 months the mortality reaches 18.7%. The factors of poor prognosis are age > 75 years, left ventricular restrictive pattern at doppler diastolic trans mitral flow evaluation, (p < 0.001), history of myocardial infarction, and renal insufficiency defined by creatinemia > 150 micromoles (p = 0.002). In conclusion heart failure with preserved left systolic ventricular function is frequent in women with hypertensive heart disease. The prognosis at mean term is better that prognosis of patients with systolic dysfunction but despite medical treatment there is a high morbidity with numerous re hospitalizations. Restrictive left ventricular filling pattern is significantly related to the occurrence of events and mortality. PMID- 12587340 TI - [Post-ischemia neurologic recovery]. AB - Stroke is one of the most common affliction of patients with neurological symptoms. Rehabilitation of stroke patients is a difficult task. Our knowledge on rehabilitation has recently improved with the emergence of data from new neuroimaging techniques. A prospective, double blind, cross over, placebo, controlled study on 8 patients with pure motor hemiparesia, is conducted to determine the influence of a single dose of fluoxetine on motor performance and cerebral activation of patients recovering from stroke. Each patient undergoes two functional magnetic resonance imaging (fMRI) examinations, one under fluoxetine and one under placebo. A single dose of fluoxetine is enough to modulate cerebral sensori-motor activation and significantly improves motor skills of the affected side. Further studies are required to investigate the effect of chronic administration of fluoxetine on motor function. PMID- 12587341 TI - [Dysferlinopathy. Example of a new myopathy]. AB - Over the past 10 years, the impact of modern microscopic pathology and molecular genetics on the knowledge of myopathies has been enormous. Dysferlinopathy is a good example. Dysferlin is a surface membrane protein without homology with known mammalian protein excepted otoferlin. It is encoded by a gene on chromosome 2. Miyoshi myopathy and limb girdle muscular dystrophy 2B have been reported to arise from defects in the same genetic locus (chromosome 2p 13). Some personal different examples are presented with typical features, high level of creatine kinase. Gene mutations, immunoblot and immunohistochemistry allow the diagnosis. Three clinical phenotypes are separated: distal myopathy, proximal myopathy, entire lower limbs posterior compartment amyotrophy. Structural changes are mild. Inflammation is not unusual and leads to the diagnosis of polymyositis. There are no correlation genotype-phenotype. PMID- 12587342 TI - [From clinical medicine to molecular genetics: the new outlook of hereditary muscular dystrophies]. AB - The introduction of molecular genetics has deeply transformed the field of hereditary neuro-muscular disorders. This is illustrated by the example of non myotonic muscular dystrophies. From five classically described entities on a clinico-pathological basis, the number of disorders identified on a genic and molecular basis is to-day about fourty. The consequences of this growing complexity should be carefully analyzed on clinical and nosographical grounds. PMID- 12587344 TI - [Prevention of skin complications from immunodepression in organ transplantation]. AB - As skin diseases after organ transplantation are often disabling and sometimes lethal, it is essential to develop preventive measures. The group "Skin and organ transplantation" of the French Society of Dermatology concentrated one's efforts on skin carcinomas and Kaposis's sarcoma. Skin carcinomas are the most frequent tumors observed in transplant recipients. Many factors participate to their pathogenesis such as genetic, environmental and iatrogenic. Their prevalence is very high in elderly transplant recipients with fair complexion, long time of immunosuppression, long time of sun exposure, multiple keratoses and warts. Preventive measures consist educational training, sun avoidance and treatment of keratoses. Transplant recipients who develop Kaposi's disease are all infected with Human Herpes Virus 8 (HHV-8). HHV-8 promoinfection occurred frequently before transplantation. Cofactors involved in the development of Kaposi' sarcoma will be probably identified by the results of an ongoing national prospective study of kidney transplant recipients. Immunosuppressive therapy should be tapered to the lowest possible level compatible with allograft function. PMID- 12587343 TI - [Three lessons learned from the first double hand transplantation]. AB - The first double hand transplantation was performed in Lyon in a 33 year-old recipient with bilateral amputation following an explosive handling accident. At 2 years, the global and the functional results were considered as very satisfactory. Three aspects are presented in this article: (1) immunosuppressive protocol was efficient and well tolerated. Only 2 skin rejection episodes on post operative days 53 and 82 were easily reversed with an increase in Prednisone doses and local applications of steroid cream; (2) successive functional magnetic resonance imaging demonstrated a global remodelling of the limb cortical map and a reversal of the functional reorganisation induced by amputation; (3) the psychological appropriation of "alien" hands improved with time. These grafts, permanently visible by the patient and others, induced a psychic defence mechanism: "denial", a lack of perception of some features of reality or a perception of reality that is immediately neglected or rejected. Denial tended to lessen as the new grafted hands gained in sensitivity and motricity. PMID- 12587345 TI - [Surgical procedures improvements thanks to navigators and robots]. AB - Navigator is a three-dimensional C-T scan image localization device, which supplies the surgeon with a real time relationship of surgical instruments with pathological lesion and vital organs. Robot controls surgical devices movements according to a various sophisticated programming, in order to palliate the imperfection of their manual use. Navigators and robots, frequently connected, allow a precise control of the instruments course, improving accuracy and efficiency of most surgical procedures. It is necessary to contribute to their improvement and development, taking in count the economic, ethic and legal problems which may be encountered. PMID- 12587346 TI - [Role, mission, and expectations of general physicians]. AB - Considering the importance of the general medicine in the French public health sector, National Academy of Medicine has created a workgroup specifically devoted to this problem (the role, the missions and the expectations of the general practitioners (GP). Several hearings of GP and of members of patients associations were organized and all the medical unions were also asked by letter. The conclusions of the workgroup mainly concern: 1) the need of better GP patients relationships 2) the need of a more appropriate professional education 3) better Hospital-GP or health insurance-GP relationships 4) improvement of the GP "status" with the possibility of a better participation to national health campaigns. Among several proposals, the members of the workgroup suggest the organisation of a national meeting devoted to the place and attempts of GP in the French public health. They consider their report as a preliminary one and they have announced that they will continue trying to make more specific suggestions. PMID- 12587347 TI - [Cognition disorders and psychotic states]. PMID- 12587348 TI - [The evaluation of cognitive dysfunctions in schizophrenia]. PMID- 12587349 TI - [Cognition and emotion in schizophrenia: neuroimaging approach]. PMID- 12587350 TI - [From neuroleptics to antipsychotics: the cognitive and emotional impact]. PMID- 12587351 TI - [Cognitive rehabilitation of psychotic patients]. PMID- 12587352 TI - [New references, new practices]. PMID- 12587353 TI - [Standard: therapeutic antipsychotic neuroleptics in acute approach: a collection of data]. PMID- 12587354 TI - [The change: subjective perception of patients treated by risperidone]. PMID- 12587355 TI - [Longitudinal comparative study of risperidone vs classical neuroleptics in the treatment of schizophrenia: 24 months of observation]. PMID- 12587356 TI - [RODOS: risperidone olanzapine drug outcome studies in schizophrenia]. PMID- 12587357 TI - [The Canadian model]. PMID- 12587358 TI - [The Swiss model]. PMID- 12587359 TI - [The Swedish model]. PMID- 12587360 TI - [Genetics, new myths and permanence of man]. PMID- 12587361 TI - Sleep disorders. PMID- 12587362 TI - Sleep and methods of assessment. AB - Sleep is a complex behavior; adequate sleep is essential for healthy functioning and even for survival. Poor sleep quality and insufficient sleep have been linked to increased risk for various illnesses, as well as with an increased prevalence of excessive daytime sleepiness and reduced quality of life. Daytime sleepiness can lead to dangerous outcomes associated with drowsy driving and has become an important public health issue. Routine health examinations that include questions about sleep habits, daytime sleepiness, and problems with sleep at night can help to educate people about the importance of good sleep habits. They provide a way to identify sleep disorders so that appropriate therapies can be instituted or proper referrals to a sleep specialist can be given. PMID- 12587363 TI - Sleep and circadian schedule disorders. AB - The timing and synchronization of human circadian rhythms is important for health and well-being. Some individuals, for reasons that remain unclear, display less resilience or flexibility in their ability to synchronize to the 24-hour world and are thus diagnosed with a circadian schedule disorder. The objective of this article is to briefly introduce concepts about human circadian timing and to review what is known about chronic, long-term circadian schedule disorders such as delayed sleep phase syndrome, advanced sleep phase syndrome, irregular sleep wake patterns, and non-24-hour sleep-wake disorder. Practical considerations for the clinician caring for these individuals are discussed. PMID- 12587364 TI - Insomnia. AB - Patients with persistent insomnia are long-suffering and in dire need of treatment. These individuals can be a challenge to treat. Duration and severity of the problem, current and prior use of sleeping medications, medical and psychiatric conditions, and experience with rebound insomnia are just some of the factors that affect treatment. Most of the insomnia problems that nurses encounter are secondary to a medical or psychiatric condition. Knowledge of assessment methods and implementation of nonpharmacological interventions can make a substantial contribution to the quality of life of persons with insomnia. PMID- 12587365 TI - Sleep disordered breathing. AB - Cause and effect relationships between sleep disordered breathing (SDB) and illness, poorer quality of life, and public health have been largely overlooked and undertreated by healthcare providers. Obstructive sleep apnea (OSA), central sleep apnea, upper airway resistance syndrome, and obesity hypoventilation are the primary syndromes that fall under the rubric of SDB. Each of these syndromes is defined; however, OSA is the most common form of SDB, and is the focus of this article. Epidemiology, pathophysiology, behavioral manifestations, cardiovascular comorbidity, clinical evaluation, and treatment for OSA are the main topics covered. The article concludes with the role of the nurse in SDB. PMID- 12587366 TI - Restless legs syndrome and periodic limb movement disorder. AB - Restless leg syndrome and PLMD are nocturnal movement disorders associated with significant adverse effects on the health and well-being of patients and their families [66]. Although the pathophysiological basis remains to be fully described, current research points to abnormalities in CNS function and neurotransmitter systems. The accurate diagnosis of RLS and PMD requires a thorough history, physical examination, diagnostic tests, and often, a referral to a sleep disorders specialist. Considering the prevalence of these conditions and their negative impact, nurses should be well-versed in the assessment and management of these problems as well as in the appropriate education of patients and their families. Nursing research is greatly needed, particularly with regard to the development and testing of biobehavioral interventions designed to decrease associated symptoms and improve clinical outcomes. Finally, because of the complexity of the clinical presentation of RLS and PLMD, this population of patients presents nurse clinicians and researchers alike with an extraordinary opportunity for interdisciplinary collaboration. PMID- 12587367 TI - Narcolepsy. AB - Narcolepsy is a chronic disorder of EDS. All patients experience EDS. Other symptoms include cataplexy, sleep paralysis, hypnagogic hallucinations, and disrupted nocturnal sleep. Treatment, usually with stimulants and low-doses of antidepressant medications, can dramatically improve the patient's quality of life. Although only advanced practice nurses may be actively involved in the diagnosis and treatment of this disorder, all nurses can encourage their patients who complain of EDS to consult a specialist in sleep disorders medicine, provide emotional support after diagnosis, and educate patients and their families about narcolepsy and its treatment. PMID- 12587368 TI - Sleep disorders in children. AB - Sleep disorders are common in childhood, and may affect multiple aspects of a child's life and the lives of other family members. A sleep disorder assessment should begin with detailed sleep history and a review of interrelated health issues. Factors contributing to disturbed sleep may be discovered or confirmed by a thorough physical examination. Thereafter, appropriate ancillary testing can provide support for a specific clinical diagnosis. The spectrum of childhood sleep disorders includes OSA, narcolepsy, RLS/PLMD, sleep onset association disorder, and parasomnias. Diagnosing sleep disorders in children remains a challenge; however, a multidisciplinary approach may provide an opportunity for productive collaboration and, thereby, more effective patient management. Centers treating pediatric sleep disorders may include providers from a variety of disciplines in pediatric healthcare, such as child psychology, pulmonology, neurology, psychiatry, nursing, and otolaryngology. Over the last decade, research in pediatric sleep disorders has expanded greatly, paralleled by an increased awareness of the importance of adequate, restorative sleep in childhood. PMID- 12587369 TI - Women and sleep. AB - Naturally fluctuating hormones (menstrual cycle, through pregnancy or menopausal transition) are not related to marked sleep disturbances in women. It is likely, however, that subsets of women will display a central nervous system vulnerability to hormonal fluctuations so that sleep disturbances manifest as a part of a complex of discomforting symptoms. Sleep is impacted directly through the circadian system or brain sleep regulation or through the development of concurrent functional changes and symptoms. Women are susceptible to sleep related disorders that are also common in men, such as primary insomnia and SBD although the contributing factors and manifestations may not be the same. PMID- 12587370 TI - Sleep and aging. AB - Synthesis of studies of sleep and aging show major increases occur in nighttime awakening and major decreases occur in sleep depth over the life span. Fatigue and drowsiness during the day, with increased napping, and earlier bedtimes, are also prevalent in the elderly. Interventions that impact these sleep variables, as well as sleep variables that change less dramatically, are needed. All indications are that the promotion of restorative sleep and daytime well-being in older adults will continue to be an important part of nursing practice affecting even more people as the population ages. Although nurses have only begun to identify and study interventions that promote sleep in nursing environments, nursing is in a position to make major contributions to the health and well-being of older adults who experience sleep disruption, especially sleep disruptions secondary to illness and aging. PMID- 12587371 TI - Birth in the United States: an overview of trends past and present. AB - The picture of birth in the United States today is complex and, as the data above indicates, difficult to describe in simplistic terms. Though many women today have come to believe that there are choices surrounding pregnancy and birth, the beliefs and practices of providers, insurers, and hospital administrators play a major role in either influencing those choices or dictating how they will be manifested. On one hand, technological advances have given women greater options with regard to the outcomes of pregnancy and birth. On the other hand, these very same technological advances place limits on the choices available to the individual. For example, increased efficiency in the placement and use of epidural anesthesia has made this a pain-control option for most of the childbearing women in the United States. The use of an epidural, however, puts limits on the choice of an institution at which to give birth and on the movements/activities of the woman during labor. Twentieth-century developments led to the almost complete demise of midwifery practice in the United States, thus taking birth away from the control of the individual woman and her close, matriarchal support system, and placing it in the hands of the patriarchal world of medicine and the institutions (i.e., hospitals) at which this approach to health care is practiced. Most births went from being normal, home-based events to becoming illness-oriented, hospital-based procedures. Just as some steps were being taken in the latter part of the twentieth century to return some of the control of birth back to pregnant women (e.g., childbirth education classes, the modern home-birth movement, increases in the number of midwifery-tended births), technological advances contributed to continued control by physicians and the hospitals of their practice (e.g., fetal monitoring, epidural anesthesia). Advances in technology have made birth possible for many individuals who otherwise would not have had the opportunity for this experience or a chance for a positive outcome (e.g., sufferers of infertility or the woman whose fetus has congenital yet repairable problems). But the widespread application of many of these advances to almost all pregnant and laboring women has raised questions as to just what is necessary for a healthy pregnancy, and are there limits to the role that technology should play in the course of what is viewed by many as a normal, nonmedical event. Just as the characters in 2001: A Space Odyssey recognized the incredible power of change that occurred in their lives as a result of the monolith being "born" into their world, so too do many women acknowledge the prodigious nature of giving birth in terms of the experience itself and the resulting effect on their lives [14]. But, as with the acknowledgment of Dr. Dave Bowman in 2001: A Space Odyssey that the technology of the fictitious future still could be wrong, thoughts of U.S. women giving birth today tend to be "in-between" a belief in the integrity of nature and a trust in the power of technology [14]. One can certainly conjecture that birth in the United States today, more than any other time in history, is at a crossroads, one in which the mostly natural, matriarchal community system of the sixteenth through nineteenth centuries is intersecting with the mostly technological, patriarchal system of the twentieth century. Will birth return to the home under the guidance of midwives, as was the case for most of America's past and as remains the case throughout much of the world? [32] Or will technology take birth to the other extreme, making it possible to eventually gestate outside the womb and rendering "delivery" a term not referring to passage through the birth canal, but to the dropping off a healthy term baby at the respective mother's doorstep? Certainly no one can predict what is to occur, but whatever that may be, hindsight will likely point to the start of the twenty-first century as the beginning of a new paradigm for birth in the United States. It should make for an interesting future. PMID- 12587373 TI - Antepartum care in the twenty-first century. AB - Major factors impacting on antepartum care in the twenty-first century were reviewed in this article, including the impact of managed care and collaborative health care teams, the malpractice crisis, antepartum visits and the quality of care, increased use of the Internet and computer technology, the increasing age of childbearing, advances in genetic and prenatal diagnosis, the importance of evidence-based medicine, the goals of Healthy People 2010 [11], the Safe Mother Initiative, and the use of complementary and alternative medicine. Recommendations for antepartum care in the twenty-first century include; the formation of integrated collaborative health care teams to deal with the effects of managed care; development of a more sensitive health care system to assist all parties involved with medical malpractice; increased expertise in the use of the Internet and computer technology; acknowledging and preparing for the increased population of older gravidas; becoming aware of advances in genetic and prenatal diagnosis; increasing the ability to critically evaluate research and incorporate well-founded study findings into clinical practice; incorporating the goals of Healthy People 2010 [11] when providing care to antepartum patients; contributing resources and manpower to the Safe Mother Initiative to decrease maternal mortality in developing countries; becoming knowledgeable, obtaining adequate training, and utilizing or performing evidence-based studies to validate the use of complementary and alternative medicine in antepartum practice; and finally to find a balance between the factors impacting on prenatal care and the ability to continue to provide safe, high-quality, continuous, sensitive, and humane antepartum care. PMID- 12587372 TI - Perspectives on the past, view of the present: relationship between nurse midwifery and nursing in the United States. AB - Relationships between U.S. nurses and nurse-midwives developed during the Progressive Era around two major points: the knowledge that lower maternal and infant mortality in England and Europe was associated with well-educated midwives, and the desire of some nursing leaders to substitute nurse-midwives for traditional midwives. In the process, traditional midwives were labeled dirty and dangerous and the practice of nurse-midwifery was marginalized. At the same time, research on nurse-midwifery showed excellent maternal and infant outcomes at the only two demonstration sites where nurse-midwifery was practiced between 1925 and 1941. This data helped the Children's Bureau and MCA push the expansion of nurse midwifery in the South and Southwest. As their numbers expanded, nurse-midwives organized nationally within the national public-health nursing association. When the association merged with other nursing organizations, however, nurse-midwives were denied their request to maintain an autonomous section on the grounds that they practiced medicine. This rejection caused a split between nursing and nurse midwifery that continues to affect interactions between the two groups of professionals. In the 1970s, the ANA, which by now supported the nurse practitioner role, embraced nurse-midwifery as an extension of nursing practice. This permitted regulation of nurse-midwifery under nurse-practice acts in all but a handful of states. Unfortunately, in the majority of these states nurse midwives were not named in statute and their practice became legally invisible. Today, many state boards require a master's of nursing for licensure even though research has shown no difference in certification exam scores between master's- and nonmaster's-prepared nurse-midwives. During the 1970s, middle-class women began to seek the services of a new group of apprentice-trained midwives, and, eventually, these midwives created their own national organization. In the early 1990s, the two national midwifery organizations met formally to craft recommendations for the education and certification of non-nurse midwives and a single standard of midwifery care. Although both organizations had developed processes for accrediting educational programs and certifying non-nurse midwives, the job of drafting a single standard of care remains incomplete. As a result, one of the major tasks facing U.S. nurse-midwives is the need to redefine their relationships with both nursing and apprentice-trained direct-entry midwives. Completion of this task holds the promise of boosting the public image of midwifery, elevating the place of midwives in the care of women and infants, and improving maternal and child health outcomes in America. PMID- 12587374 TI - Intrapartum care in the twenty-first century. AB - Women will continue seeking obstetrical care from nurses, midwives, and physicians throughout the twenty-first century. In many areas of the country, they will be able to find a midwife who will assist them in having a very personal birth experience. The ACNM remains committed to producing more midwives. More midwives may mean that practitioners educated in normal pregnancy will attend the vast majority of normal births, freeing physician colleagues to best use their skills and expertise in caring for women with medical and obstetrical problems. As most midwives are likely to continue working in hospital settings, those settings will continue to change, offering women more of the comforts and amenities of home. Home birth and water birth may continue to be available with midwives in attendance, though the forces of economics and insurers may restrict the availability of these options for women. Women desiring care in a birth center may find it difficult to locate one within a reasonable distance. The in hospital "birthing suite", with a midwife in attendance, will be the most likely setting for the vast majority of midwifery attended births. A collection of more evidence through research will stir debate amongst health care providers. Increased access to that information will bring consumers into the debate as well. In the twenty-first century, information will be a very powerful force of change in obstetrical health care. In recent years, legal liability and economics have strongly influenced obstetrical practice. Though this may continue to be true, the impact of more evidence on which to base practice, and the new access that women have to that information, will undoubtedly affect the way care is delivered. A central slogan of the ACNM is "Listen to Women". That will happen more than ever in the twenty-first century. There will be more midwives, more evidence to support midwifery care, and more women learning that birth can and should be a personal, healthy, and empowering experience. These women will seek midwives who practice wisely, blending science with art and intuition. They will learn that the childbirth wisdom that has been passed down through the ages, from woman to midwife to healer to nurse and to midwife, again, delivers the birth experience back to the mother and the healthy baby to the world [3,7]. PMID- 12587375 TI - Technology in the birthing room. AB - The use of technology is not benign. As with any health care intervention, there are associated risks and benefits. The practitioner needs to constantly consider the benefits of the technology versus the naturalistic birth experience. The use of technology should optimize birth outcomes while maintaining a balance that provides for the best possible human birth experience. Technology, however, does have merit in the birth setting, regardless of location, but its use should be evaluated on an individual, as needed, basis. The most common technological advances currently available for assessment and maternal/fetal care during birth include electronic fetal monitoring, ultrasonography, blood pressure screening, maternal/fetal pulse oximetry, and infusion pumps. All obstetrical care providers must be familiar with the forms of technology currently available and be aware of emerging technologies for use during the birthing process. PMID- 12587376 TI - Politics in the birthing room. AB - Women have always braved their way through childbirth one way or another. The hands surrounding them as they make their journey have offered many things. For some it has been herbs or a totem to hold. For others, it has been a tether to a beeping machine or a drug to transform the experience. No one has yet found the method(s) that addresses every pregnant woman's fear or pain during labor and birth. And no one can truly guarantee that all will be okay. The contemporary medical profession offers to shoulder the burden for women by "taking care of it for them," The managed care industry assures the public that they will provide pregnant women with the highest quality of care and the greatest decision-making power. Elected officials promise that they will save women and newborns from unscrupulous legal practices and from the abuses of managed care. But these goals, for better or worse, have yet to be fully realized. No matter the circumstances, women will continue to have babies. Their labors will be short or long, will occur surrounded by flowered decor or in sterile wards, and will be attended by physicians who are overburdened by the demands of maintaining a practice or by a midwife who must battle with institutional protocols to care for her clients. The battle for the political victories in women's health care will undoubtedly continue for some time to come. The strength and resolve of women and their families will determine the outcome and give voice to the consumer and the midwife in the board-room, in the hearing room, and in the birthing room. PMID- 12587377 TI - Midwifery education today. AB - Today, there are more than 7000 CNMs and CMs in the United States, who attend approximately 9% of American births annually in hospitals, birth centers, and homes. Midwives work in a variety of practice models, including group practices with physicians, HMOs, private practices, rural and urban community health centers, and large managed care organizations. CNMs and CMs also serve as administrators, policy makers, and consultants in international maternal and child health. Midwifery education has come a long way since 1931, when the first education program started. The dedication of CNMs to our clients and the midwifery model of care, as well as these midwifery educational innovations, will continue to support the profession's goals of offering humane, holistic, and safe health care to women and their families. PMID- 12587378 TI - [Chemoradiotherapy in the adjuvant treatment of gastric adenocarcinomas: real progress?]. AB - Frequency of local and distant failures after gastrectomy has led to extended lymph nodes dissection to obtain a better locoregional control. However, five year survival rates were not significantly different between patients undergoing D2 and D1 lymphadenectomy, and higher morbidity and post operative deaths were reported in large randomised trials (respectively 25% vs 48% and 4 vs 13%). Additionally, several metanalysis failed to demonstrate a significant survival advantage with adjuvant chemotherapy. The results of the first trial demonstrating one advantage to adjuvant post-operative chemoradiotherapy should modify the standard care. Disease free and overall survival after surgery alone and after surgery and concurrent chemoradiotherapy were respectively 31% vs 48% and 41% vs 50%. The intergroup trial demonstrate that better local control improve survival if radiation fields include stamps, tumour bed, proximal nodal chains and nodes corresponding to D2 extended lymph nodes dissection. Treatment was feasible with few severe toxic effects (1%). Of the 281 patients, 17% stopped treatment because toxic effects. Technical modalities of radiotherapy and post operative nutrition support, which are critical points of interest for this treatment, are also discussed. PMID- 12587379 TI - [Principal therapeutic uses of monoclonal antibodies in oncology]. AB - In the past few years, new agents based on monoclonal antibodies have been developed in Oncology. Indeed in some case tumor cells express antigenic targets at higher levels than normal cells. There are 2 main types of monoclonal antibodies that can be either conjugated to cytotoxic drugs or radio-active compounds or be non-conjugated. Among the last category, some are currently used in the treatment of patients, including 2 monoclonal antibodies targeting receptors with tyrosine kinase activity (HER2: Herceptin (DCI: trastuzumab), EGFr: Erbitux (DCI: cetuximab). A third monoclonal antibody is commonly used in cancer treatment, which targets CD20, a transmembrane marker of B lymphoma (MabThera (DCI: rituximab). Both Herceptin and MabThera have been associated with improved survival in patients with breast carcinoma and lymphoma, respectively. New promising agents are under investigation such anti EGFr in colon and head and neck carcinoma or new compounds such as anti-VEGF. These examples outline the importance of the recent progress in selectively targeting tumor antigen and the potential impact of these approaches in Oncology. PMID- 12587380 TI - [Target-volume selection and delineation in the cervico-maxillo-facial region: beyond the concepts of the ICRU]. AB - Improvement in irradiation techniques, which allows dose distributions sculpting around volumes of very complex shapes, has revealed the limitations in selection and delineation of target volumes. The use of functional imaging (PET, fMRI) in addition to anatomic imaging, will probably bring an extra level of complexity to this issue. In particular, the use of specific markers to visualize biological pathways known to influence response to ionizing radiation (e.g. hypoxia, proliferation) could lead to the delineation of sub-target volumes for delivering an extra boost dose. Such concept of Image-Guided Radiation Therapy still need to be tested in experimental models and in well defined clinical situations before its use in a routine clinical set-up. PMID- 12587381 TI - Dose conformation in IMRT for head and neck tumors: which solution to apply? AB - At Ghent University Hospital, IMRT for head and neck cancer is routinely performed. The desired dose distribution is defined upfront as a range of acceptable doses assigned to each voxel of volumes of interest. It was found important to specify the range of acceptable doses separately to areas of the PTV either in or outside the buildup zone as well as to areas which do or do not intersect with PTV-dose limiting organs at risk (OAR). To avoid high doses at distance from the PTV, the creation of a "surrounding" OAR which is the whole scanned volume minus the PTV was found efficient, especially if inside this OAR, subvolumes were created at increasing distance from the PTV. By specifying inside these subvolumes maximum dose constraints which decreased with distance from the PTV, conformality is secured. The creation of these additional PTV and OAR subvolumes allows comprehensive and unambiguous definition of the range of acceptable doses and thereby avoids user-interactive assignment of weights to the terms of the objective function during optimization. The efficiency of inverse planning is highly improved. Its outcome is predictable, plan evaluation is objective as the plan either does or does not comply with the predefined range of acceptable doses. Accurate reporting of the planned dose distribution is facilitated by description of the dose range to all volumes. The expense of this procedure is modest and lays mostly 1) in the creation of the subvolumes, which can be done semi-automatically by modern image segmentation tools and 2) in the inclusion of constraints to all subvolumes into the objective function. PMID- 12587382 TI - [Upper aerodigestive tract cancers: clinical benefits of conformal radiotherapy and intensity modulation]. AB - The conformal radiotherapy approach, three-dimensional conformal radiotherapy (3DCRT) or intensity-modulated radiotherapy (IMRT), is based on modern imaging modalities, efficient 3D treatment planning systems, sophisticated immobilization systems and rigorous quality assurance and treatment verification. The central objective of conformal radiotherapy is to ensure a high dose distribution tailored to the limits of the target volume while reducing exposure of normal tissues. These techniques would then allow further tumor dose escalation. Head and-neck tumors are some of the most attractive localizations to test conformal radiotherapy. They combine ballistic difficulties due to particularly complex shapes (nasopharynx, ethmoid) and problems due to the number and low tolerance of neighbouring organs like parotids, eyes, brainstem and spinal cord. The therapeutic irradiation of head-and-neck tumors thus remains a challenge for the radiation oncologist. Conformal radiotherapy does have a significant potential for improving local control and reducing toxicity when compared to standard radiotherapy. However, in the absence of prospective randomized trials, it is somewhat difficult at present to evaluate the real benefits drawn from 3DCRT and IMRT. The published clinical reports on the use of conformal radiotherapy are essentially dealing with dosimetric comparisons on relatively small numbers of patients. Recently, a few publications have emphasized the clinical experience of several precursor teams with a suitable follow-up. This paper describes the current state-of-the-art of 3DCRT and IMRT in order to evaluate the impact of these techniques on head-and-neck cancers irradiation. PMID- 12587383 TI - [Target-volume contours in radiotherapy. General principles]. AB - Target volume delineation remains an important medical step for conformal radiotherapy. In this general presentation are discussed: the main difficulties for evaluation of gross tumor (GTV) and clinical target volume (CTV), the main recommendations for representation according to ICRU 62 report, differences observed during inter-observer volume comparisons. PMID- 12587384 TI - [Basic anatomopathological features of tumors and their extensions. Prognostic and therapeutic implications]. AB - Surgical pathology analysis of biopsies or surgical specimens directly determines the diagnosis of the disease. It gives also the different characteristics that are useful for the establishment of the prognosis and for the treatment of each tumor. We report here a synthesis of the different characteristics of neoplastic lesions, necessary or useful for the establishment of the care program of the patients. These macroscopic and microscopic analyses should result in the most precise possible diagnosis and should take into account a list of other factors that allow the development of an optimal strategy for therapy, and the establishment of prognosis of the tumor. These characteristics include properties of the tumor like the size, shape, exact location, and the grade of the lesion, etc. They also concern the relationship of the tumor with certain structures like the basal membranes, the extracapsular extensions, the stroma reaction, the lymphatic vessels, the nerves, etc. The description of local and loco-regional spreading, as well as the analysis of the surgical margins gives also essential information. Sometimes, the description of modifications induced by neoadjuvant therapy, as chemo or radiotherapy, allows the evaluation of the efficiency of these treatments. PMID- 12587385 TI - [Computerized tomography for dosimetry purposes: technical factors in helical acquisition. Importance and limits of this technique in target-volume delineation and dose calculation]. AB - The computerized tomography (CT) used for target volumes contouring is an essential step of the radiotherapy process. The CT technical factors must be adapted to explored organs and anatomic regions: data acquisition parameters and reconstruction algorithm, digestive or intravenous contrast. The choice of these parameters and its consequences for contouring are discussed. Respiratory motion must be considered for some organs. Helical CT allow a data acquisition during one apnoea. Knowledge of these technical factors is essential for good contouring and for the future use of multiple row detectors. Precise exams protocols depending on lesions, on treatment methods and patients are demanded. Close cooperation between radiation oncologists, physicists and radiologists is not only a wish but becomes a necessity. PMID- 12587386 TI - [Target-volume and critical-organ delineation for conformal radiotherapy of prostate cancer: experience of French dose-escalation trials]. AB - The delineation of target volume and organs at risk depends on the organs definition, and on the modalities for the CT-scan acquisition. Inter-observer variability in the delineation may be large, especially when patient's anatomy is unusual. During the two french multicentric studies of conformal radiotherapy for localized prostate cancer, it was made an effort to harmonize the delineation of the target volumes and organs at risk. Two cases were proposed for delineation during two workshops. In the first case, the mean prostate volume was 46.5 mL (extreme: 31.7-61.3), the mean prostate and seminal vesicles volume was 74.7 mL (extreme: 59.6-80.3), the rectal and bladder walls varied respectively in proportion from 1 to 1.45 and from 1 to 1.16; in the second case, the mean prostate volume was 53.1 mL (extreme: 40.8-73.1), the volume of prostate plus seminal vesicles was 65.1 mL (extreme: 53.2-89), the rectal wall varied proportionally from 1 to 1, 24 and the vesical wall varied from 1 to 1.67. For participating centers to the french studies of dose escalation, a quality control of contours was performed to decrease the inter-observer variability. The ways to reduce the discrepancies of volumes delineation, between different observers, are discussed. A better quality of the CT images, use of urethral opacification, and consensual definition of clinical target volumes and organs at risk may contribute to that improvement. PMID- 12587387 TI - [Cancer of the rectum: target volumes for preoperative radiotherapy]. AB - Preoperative radiotherapy is the standard treatment for locally advanced resectable rectal adenocarcinoma. The total mesorectal excision leads to a dramatic increase of local control rate. Thus, the mesorectal space is the usual field for the spread of rectal cancers cells. It could therefore be considered as the clinical target volume (CTV) in the preoperative conformational radiotherapy. From the anatomical basis and radiological contributions, we propose several CTVs for different locations of rectal carcinoma. PMID- 12587388 TI - [Preoperative chemotherapy and chemoradiotherapy for non-small-cell bronchial cancers]. AB - The very disappointing results obtained by surgery in resectable non-small-cell lung cancer have led to a high active clinical research concerning pre- or postoperative treatment. Preoperative treatment has several distincts goals: to increase survival for patients suitable for surgery, to limit surgery or transform borderline or non resectable cancer into resectable tumors. Available datas on preoperative treatments for non-small-cell lung cancer provide from three types of therapeutics trials: 1/Some phase II studies of neoadjuvant chemotherapy have demonstrated that the neoadjuvant approach was feasible, and didn't compromise surgery. 2/Phase II trials of neoadjuvant chemoradiotherapy, performed for the majority on more extensive cancers, have demonstrated that this approach was also feasible at the expense of higher but still tolerable toxicity. 3/Phase III randomised published trials exclusively deal with preoperative chemotherapy with different results: two of them concerned a small number of patients presenting with non-small-cell lung stage IIIA cancer: they are positive. The third concerned 373 patients presenting with stage I, II, IIIA cancer: the three-year survival was increased by 11%, but this difference is not yet significant. The benefit essentially appeared for stage I and II. One trial comparing preoperative chemotherapy and radiochemotherapy has been reported, concluding to the superiority of the association. These observations suggest that the clinical research should now be different for stages I and II, and stage IIIA. PMID- 12587389 TI - [Surgical morbidity for bronchial cancers after neoadjuvant treatment]. AB - Two randomized trials, in 1994, have demonstrated the benefit of neoadjuvant chemotherapy, in term of median survival, for stage III lung cancer. Further studies have evaluated the potential benefit of chemotherapy or chemotherapy radiotherapy association, either for patients suitable for surgery, or for non resectables tumors. However, these treatments treatments may increase the morbidity of surgery. Such an increase has not been demonstrated, except in one study, after chemotherapy alone before surgery. When radiation therapy is associated with pre-operative chemotherapy, the risk of complications seems to be dose dependent: low for doses below 50 Gy, important for doses over 55 Gy. These datas justify pre-operative lung function measurements and modifications of the surgical technic, especially for the lymphadenectomy extension. Despite this potential increase of morbidity, the benefit of neoadjuvant treatment is real. PMID- 12587391 TI - [Non-small-cell bronchial cancers: improvement of survival probability by conformal radiotherapy]. AB - The conformal radiotherapy approach, three-dimensional conformal radiotherapy (3DCRT) and intensity-modulated radiotherapy (IMRT), is based on modern imaging modalities, efficient 3D treatment planning systems, sophisticated immobilization devices and demanding quality assurance and treatment verification. The main goal of conformal radiotherapy is to ensure a high dose distribution tailored to the limits of the target volume while reducing exposure of healthy tissues. These techniques would then allow a further dose escalation increasing local control and survival. Non-small cell lung cancer (NSCLC) is one of the most difficult malignant tumors to be treated. It combines geometrical difficulties due to respiratory motion, and number of low tolerance neighboring organs, and dosimetric difficulties because of the presence of huge inhomogeneities. This localization is an attractive and ambitious example for the evaluation of new techniques. However, the published clinical reports in the last years described very heterogeneous techniques and, in the absence of prospective randomized trials, it is somewhat difficult at present to evaluate the real benefits drawn from those conformal radiotherapy techniques. After reviewing the rationale for 3DCRT for NSCLC, this paper will describe the main studies of 3DCRT, in order to evaluate its impact on lung cancer treatment. Then, the current state-of-the-art of IMRT and the last technical and therapeutic innovations in NSCLC will be discussed. PMID- 12587390 TI - [Locally advanced non-small-cell bronchial cancer: role of exclusive chemoradiotherapy]. AB - Lung cancer is the most frequent form of malignant tumours. The prognosis is poor with a 5-year cure rate which increased from approximately 6% in the sixties to only 15% in the nineties. Surgery remains the reference treatment but only a small minority of patients (about 25%) present with operable disease. The post surgical 5-year survival is only 25%, providing the rationale for the current research on adjuvant treatments for control of both local and metastatic disease. In that context, the combination of radiotherapy and chemotherapy, commonly referred to as chemo-radiotherapy, has assumed considerable importance: either exclusively in inoperable patients (inoperable tumour or patients inoperable for medical reasons), or pre-operatively. This article reviews the results of the pivotal definitive chemoradiotherapy studies in non-metastatic non-small-cell lung cancer. With exclusive chemoradiation, the concomitant scheme seems to be the most favourable, results issued from randomised studies are expected to confirm that point. An increased toxicity is observed, and the advent of conformal therapy may allow another survival gain. Optimal treatments integration will be necessary. PMID- 12587392 TI - [Thoracic radiotherapy and control of respiration: current perspectives]. AB - Three-dimensional conformal radiotherapy (3D CRT) is adversely affected by setup error and organ motion. In thoracic 3D CRT, breathing accounts for most of intra fraction movements, thus impairing treatment quality. Breath control clearly exhibits dosimetric improvement compared to free breathing, leading to various techniques for gated treatments. We review benefits of different breath control methods--i.e. breath-holding or beam gating, with spirometric, isometric or X-ray respiration sensor--and argument the choice of expiration versus inspiration, with consideration to dosimetric concerns. All steps of 3D-CRT can be improved with breath control. Contouring of organs at risk (OAR) and target are easier and more accurate on breath controlled CT-scans. Inter- and intra-fraction target immobilisation allows smaller margins with better coverage. Lung outcome predictors (NTCP, Mean Dose, LV20, LV30) are improved with breath-control. In addition, inspiration breath control facilitates beam arrangement since it widens the distance between OAR and target, and leaves less lung normal tissue within the high dose region. Last, lung density, as of CT-scan, is more accurate, improving dosimetry. Our institution's choice is to use spirometry driven, patient controlled high-inspiration breath-hold; this technique gives excellent immobilization results, with high reproducibility, yet it is easy to implement and costs little extra treatment time. Breath control, whatever technique is employed, proves superior to free breathing treatment when using 3D-CRT. Breath control should then be used whenever possible, and is probably mandatory for IMRT. PMID- 12587393 TI - [Intracerebral radiotherapy under stereotaxic conditions]. AB - Stereotactic radiosurgery is used for treating several brain diseases. Radiosurgery is a non-invasive alternative to surgery for brain metastases, and randomized trials are on going to assess the role of radiosurgery. Radiosurgery has been advocated for patients with small benign meningioma or with vestibular schwannoma, but there is no proof of efficacy and safety of radiosurgery in comparison with other treatments. Radiosurgery can obliterate 80-90% of small arteriovenous malformations, but no information exists on the survival of treated compared with untreated patients. The limited information available suggests that radiosurgery should be fully evaluated in well-designed prospective studies. PMID- 12587394 TI - [Probabilities of controlling tumors and complications (TCP/NTCP) after radiotherapy: methodologic, physical, and biological aspects]. AB - Radiotherapy is aimed at getting the best possible therapeutic ratio (tumor local control versus morbidity). Physicists and radiation oncologists have to evaluate explicitly or implicitly the probability of induced complications to normal surrounding tissues. This is based on published data and clinician's experience. Quantitative methods have been introduced with different models in order to predict the impact of partial or global irradiation on a normal organ. These models correspond to the Tumor Control Probability (TCP) and Normal Tissue Complication Probability (NTCP). These biological models may be useful to evaluate the quality of a treatment planning or for the optimization process. The methodologies used and the clinical data are developed and discussed. PMID- 12587395 TI - [Accidents in radiotherapy: historical account]. AB - Radiotherapy accidents are exceedingly rare. However, they may have major negative consequences: for health (and sometimes life) of victims as well as for the trust that patients put in radiotherapy and radiation oncologists. Each accident must be pointed out, analysed and reported, in order to allow preventive actions, avoiding repetitive accidents. Through examples of majors accidents occurred all over the world in the last decades, affecting professionals, public or patients themselves, the necessity of transparency is demonstrated. The International Commission of Radiobiological Protection has drawn positive lessons from such accidents and insists on following recommendations: necessity of sufficient number and competent professionals, importance of continuous and initial education, information of professionals and, in general, a strict Quality Assurance program. It is clear that each radiotherapy center remains at risk for errors. It is essential to develop preventive procedures to avoid transformation of errors into accidents. In that context, complete and detailed description and reports of each anomaly or incident must be encouraged as it is done for sectors of aviation or nuclear industry. Radiation oncology must develop such a culture of transparency and of systematic report of all incidents. PMID- 12587396 TI - [Quality control of equipment used for radiotherapy]. AB - Due to the modifications of components, to unexpected breakage of elements or to electronic dysfunctions, the performance of radiotherapy machines may decrease with age. Quality Assurance procedures and maintenance program are necessary to guarantee the performances. For linear accelerators, modus operandi of control tests and their frequency are based on regulations and recommendations widely published, that are presented here. Concerning accessories, especially those recently developed (multileaf collimators, dynamic wedges, ...), recommendations remains to be defined. Simple tests are proposed. Concerning numerical imaging systems, widely used for three dimensional dosimetry, image quality and geometry controls must be performed with phantom tests. For portal imaging, a quality assurance program is proposed. A strict and complete Quality Assurance program is essential to guarantee quality and safety of the treatment. A regular control of linear accelerator is one of the important component of this program. It suppose the implementation of permanent tests procedures, periodically modified following technological progresses and treatment techniques. Measurements must be sensible to variations below the tolerance level defined during the installation process. The analysis of the variations of measurements with time are an objective criterion of quality. PMID- 12587397 TI - [Safety in radiotherapy: Control of software and informatics systems]. AB - The development of computerized systems in radiotherapy opens new challenging possibilities. There is however a major risk of specific errors if the use of such systems is not properly controlled. On the other hand, the software sophistication, such as encountered in modern treatment planning systems (TPS), makes it very difficult to set up efficient quality assurance programs compatible with the workload of users in charge of daily routine. From the analysis of previous accidents involving TPS, we can conclude that the major risks are erroneous calculations of monitor units resulting from a poor understanding of the software capabilities combined to a lack of vigilance with respect to the data obtained from a computer. To reduce the risks, it is necessary to setup a comprehensive quality assurance program covering all steps of the process, including commissioning of a new system and systematic checks of individual treatment plans. Such a program offers the users the opportunity to acquire a deeper understanding of the system and avoid potential pitfalls. There is presently a lack of consensus for an international protocol related to quality assurance of computerized systems in radiotherapy. It is therefore important to define protocols at the national level, with special consideration of the aspects which are more likely to generate significant errors. PMID- 12587398 TI - [Recommendations for quality control in curietherapy]. AB - Brachytherapy consists of sealed radioactive source implantation. The diversity in the nature of radioelements, in their energy and activity requires strict implantation and utilization rules. These rules include radioactive source physical parameters check, after-loading machine and treatment planning system quality assurance and safe and reproducible dosimetric systems. Patient and medical workers information guarantee radioprotection and prevention of accidental exposures. PMID- 12587399 TI - [The patient care experience in radiotherapy: perspectives for better patient support]. AB - This paper reports the patient care experience during the course of the radiotherapy. Plights are multiple. Patients are confronted with a complex and unknown distressing space, an irrelavant information, a banalization of side effects, an isolation with a frequent inadequate support of their family or the caregivers team, with the fear of a definitive abandonment at the end of the treatment without comforting follow-up. It is imperative to state a real policy in order to improve the patient support. Sensibilization and training of the caregivers, in spite of a frequent overbooking technical work, is required in a pluridisciplinary approach to provide a relevant reception with the collaboration of psychologists, social workers and self care groups and associations. The personal implication of the physicians and technologists is also essential. The simple smile is the intangible proof of the reliable emotional support. PMID- 12587400 TI - [Psycho-oncology: methods, uses]. AB - Psycho-oncology is exercised daily in the cancerological care in concert with other practices. It is one of the component of the pluridiscinarity. Preventive actions towards patients and loved ones result from this practice as well as the unrelenting seeking of psychological difficulties. In addition, psycho-oncology enhances the coordination of patients care. Integrated to the "supportive care" it allows a better understanding of the psychosocial dimension in the decision process. Constant collaboration with other medical practitioners ensures awareness and a reciprocal training, which are at the core of the care relationship. PMID- 12587401 TI - [The organization of psycho-oncology]. AB - Many specialists are acting for psychological support: oncologists, psychologists, psychiatrists. Their respective role and the necessary good adjustment between actors are discussed. Quantitative requirements depends on number of patients and dispersion of medical units. The insufficient number of oncological staff cannot be compensated by the psycho-oncological team whose action is complementary of the technical staff support. The psycho-oncological team may be integrated in the oncology unit or autonomous. A network organisation is still uncommon. The intervention of external units of psychiatry, not specialised in oncology, is a less satisfying solution. Actually, 25% of the 94 French centres have neither psychiatrist nor psychologist. The same investigation has shown that 20% of patients request a consultation with a psychiatrist or a psychologist, but only half of them obtain it. There is also a need of formation as well for psychiatrists and psychologists as for oncologists. All that is necessary to satisfy the demand of support of patients and of their family. PMID- 12587402 TI - [Metamorphosis of the uterus from Hippocrates to Ambroise Pare]. AB - The treatise Des monstres et prodiges (1579, 1585) by Ambroise Pare includes a vignette depicting a monstrous embryo in the form of a human head surrounded by snakes. This picture belongs to the iconographic tradition relating to the Graeco Roman mythology of sexuality and procreation. It derives from the belief in the womb's animal nature, illustrated on magic Graeco-Roman and Byzantine gemstones, where the uterus is shown in turn as a cupping vessel, a scarab-beetle, an octopus or the head of Gorgo. PMID- 12587403 TI - [Melancholia and reflection on the genius of Huarte de San Juan. Temperament and powers of the soul in Galenism of the Spanish Renaissance]. AB - Huarte de San Juan's Examen de ingenios (Examination of men's wits) of 1575 is one of the most significant Spanish contributions to Renaissance thought. The treatise reached an enormous popularity in Europe during the 16th and the 17th century as demonstrated by numerous translations in Italian, French, Portuguese, English, German or Dutch. However, it is barely known in modern times. Huart's doctrine illustrates the Renaissance attempt to restore the purity of ancient medicine, while proposing a "scientific" basis for the project of a systematic professional orientation. By means of the ancient theory of humours and that of the philosophical reflection on the genius, Huarte articulates a new pattern for the intellectual ability, seen as highly individual, but also deeply rooted in the biological nature of the human being The intellectual fecundity is explained by recourse to humoural physiology with a special focus on the traditional image of the melancholic temperament, redefined in the spirit of the pre-modern anthropology of the variety. PMID- 12587404 TI - [Drug therapy of psychiatric patients in the middle of the 19th century: the drug armamentarium of Ludwig Binswanger sen. in his "Asyl Bellevue" ]. AB - This paper examines the drugs used in Ludwig Binswanger's private sanatorium "Bellevue" in Kreuzlingen on the Lake of Constance between 1857 and 1870. The available patient records have revealed the administrated drugs, prescriptions have been transcribed, and the guidelines for the use of the drugs have been reconstructed by means of the literature. It becomes obvious that the armamentarium of drugs prescribed was limited and can be connected with the treatment of physical ailments which the "somatist" Binswanger considered as the main cause for psychic disorders. Binswanger was very careful when prescribing one of the few available "psychotropic" drugs like opium and morphine. This restraint in using drugs is in contrast with many recommendations and habits of Binswanger's contemporaries and shows that he put higher emphasis on the effectiveness of the "therapeutic milieu" than on a pharmacotherapy of insanity. PMID- 12587407 TI - Building patient satisfaction. PMID- 12587405 TI - [Owsei Tempkin (1902-2002). Medical historiographer of the 20th century]. AB - An overview is given of the life and work of Owsei Temkin (1902-2002), historian of medicine in Leipzig and Baltimore. PMID- 12587408 TI - Listen closely. Communication helps collaboration with the O.R. PMID- 12587409 TI - Operating information. PMID- 12587410 TI - Finding out who's naughty and nice. PMID- 12587411 TI - Open findings. Resulting statistics of SUD survey. PMID- 12587412 TI - [The sticky platelet syndrome: a frequent but unrecognized cause of thrombophilia]. PMID- 12587413 TI - [Quality of life after ileo-anal anastomosis]. AB - BACKGROUND: The ileo-anal pouch is the surgical procedure of choice for patients with Ulcerative Colitis or Familial Adenomatous Polyposis, but has functional limitations such as a higher frequency of bowel movements, anal leakage, and sometimes the necessity of a protective anal pad. OBJECTIVE: To analyze the functional results and quality of life after the pelvic pouch. MATERIAL AND METHODS: This is a descriptive, prolective, and cross-sectional study that analyzes the clinical variables, functional results and self-reported quality of life of patients after an ileo-anal pouch. A correlation between postoperative clinical variables and quality of life was searched. RESULTS: Twenty-seven patients were included. Mean age was 36 years. Surgical indications for the ileo anal pouch were Ulcerative Colitis in 17 (63%), Familial Adenomatous Polyposis in 9 (33%) and a colo-rectostomy stricture in 1 (4%). Mean number of bowel movements was 4 at day and 1 at night. Eighteen percent of patients referred anal leakage, 11% had pouchitis, and 11% small bowel obstruction. Most of the patients reported high scores in all evaluated quality of life scales. There was a correlation between lower scores of quality of life and a higher number of bowel movements. CONCLUSIONS: The majority of patients reported an adequate quality of life after the ileo-anal pouch; there was a correlation between lower scores of quality of life and a higher number of bowel movements. PMID- 12587414 TI - [Accuracy and diagnostic utility of the Hospital Anxiety and Depression Scale (HAD) in a sample of obese Mexican patients]. AB - The Hospital Anxiety and Depression Scale (HAD) have been used in Mexico in drug abusers, burned patients, older people, with renal insufficiency and high-risk pregnant women. The aim of this study was to determine reproducibility and accuracy of the questionnaire in a sample of obese subjects. A group of 75 obese patients (BMI > 27) without diabetes mellitus were invited to participated in the study. Diagnosis of anxiety or depression was made by an structured interview based on the DSM-IV criteria, and they were requested to complete the HAD. All subjects were randomized for the manoeuvre sequence. Sensibility specificity, positive predictive value and negative value, and unweighted kappa coefficient (for concordance) were calculated for the two procedures. The questionnaire reproducibility was assessed buy test-retest with other 25 independent subjects. Internal validity was estimated by alpha Cronbach, Guttman and intraclass correlation coefficients. Mean age was 39.7 +/- 11.5 years and BMI 39.1 +/- 9.6. The best cut off point for anxiety was 8 points (Kappa 0.68) and for depression 7 points (Kappa 0.73). Mean age for test-retest was 39.2 +/- 14.5 years and BMI 45.3 +/- 14.6. The alpha-Cronbach was 0.84 for the first tes. and 0.86 for the second. Intraclass coefficient correlation was 0.946. The HAD is applicable for obese subjects, it is reproducible and concordant with a structured interview. PMID- 12587415 TI - Detection of human influenza virus in Yucatan, Mexico. AB - OBJECTIVE: Influenza virus is the most common cause of Acute Respiratory Infections (ARI) world wide. In patients with chronic condition, infection by the influenza virus can cause complications such as pneumonia which may have fatal outcome. The aim of this work was to determine the frequency of human influenza virus in outpatients with influenza-like illness (ILI) and in those patients admitted to hospital with community acquired pneumonia (CAP) in Yucatan, Mexico (October 1998-July 1999). MATERIALS AND METHODS: Throat swabs were collected from ILI and CAP patients and processed to detect respiratory viruses. All clinical samples were tested for seven respiratory viruses using a rapid indirect immunofluorescence test (IFI). Clinical samples with positive results for influenza virus by IFI were inoculated into chick embryo eggs and/or MDCK cells for viral isolation. All influenza virus isolates were typed using the WHO influenza Kit 1998-1999. RESULTS: A total of 288 clinical samples were collected. Influenza virus type A was diagnosed in 29 clinical samples (10%), no other respiratory viruses were identified. Influenza virus was present with 8.9% (17 out of 189) in ILI patients, whereas with 12.12% (12 out of 99) in CAP patients. Influenza virus was detected from December to July. Six viral isolates were obtained and identified as influenza A (H3N2). CONCLUSION: Human influenza virus is certainly a cause of ARI and pneumonia in Yucatan, Mexico. The results showed that influenza virus contributes to at least 8.9% of the ARI, and more importantly to 12% of CAP patients. Positive cases were present in a different pattern to temperate zones where the peak of incidence occurs during autumn and winter. PMID- 12587416 TI - [Frequency of apolipoprotein E in a Nahua population]. AB - The presence of different ethnic groups in Mexico may give rise to genetic diversity between the native Indian population and the Mestizos. It is therefore of medical and anthropological interest to analyze the genotypes of disease associated loci, such as polymorphism in the apolipoprotein E gene, whose 4/4 allele increases the risk of Alzheimer's disease and coronary heart disease in other populations. We studied a Nahua Indian-population in the State of Morelos (Santo Domingo Ocotitlan). The ABO blood type of all individuals was determined and compared with the findings of other Nahua group from the State of Puebla. Without statistical significant differences in O, A and AB groups between both populations (p > 0.05). The allelic and genotypic frequency of apolipoprotein E was similar to that observed in other Mexican indian (Mazatecans, Mayans) and Mestizo populations, however there was a statistically significant difference when the results were compared to the allelic frequencies of other Amerinds: The Cayapa (Ecuador) for the epsilon 3 and epsilon 4 alleles (p < 0.002); the Nuuk (Greenland) for epsilon 3 and epsilon 4 alleles (p < 0.0001 and p < 0.002 respectively); and the Ammssalik (Greenland) for both alleles with p < 0.0001 and p = 0.04 respectively. In the case of the genotypes, there was statistically significant difference for the 4/3 genotypes, but a non significant difference for the 4/4 genotype. This is a descriptive study which contributes to the knowledge of the genetic structure of Mexican population. PMID- 12587417 TI - [Opinions of a group of university students about science and technology]. AB - OBJECTIVE: To learn the opinions of university students of four different areas on the impact of science and technology on society. SUBJECTS: One Hundred and sixty three close to graduate students of the Universidad Autonoma Metropolitana campus Iztapalapa, distributed as follows: Administration 59, Biology 50, Social Sciences 36 and Engineering 18. METHODS: For the survey we translated into spanish part of a questionnaire employed in several countries to explore ideas on the impact of science and technology on society of several groups. It contained general questions such as. Do you believe that science and technology are equally good or bad to society, or degree of knowledge of several technologies such as computation or in vitro fertilization. It includes also more specific questions, such as would your have problems with the use of genetically modified vegetables? RESULTS AND DISCUSSION: The results suggested that Administration and Social Sciences students had less interest in Science and Technology than the other, and that in general, the knowledge of all students is rather limited including biotechnology, genetic enginering and gene therapy. We compared the results with those obtained previously in a group of Mexican Physicians and Biology students from India, Thailand and Singapor. PMID- 12587418 TI - [In vitro evaluation of the chemopreventive potential of saffron]. AB - Cancer is a very important national health problem in Mexico, while a significant increase in the total and childhood cancer mortality has been recorded during the last decades. Chemoprevention, defined as the use of natural or synthetic agents to prevent or to block the development of cancer in human beings, is a new and promising strategy in the battle against cancer. Saffron, obtained from the dried red-dark stigmas of Crocus sativus L., an important spice rich in carotenoids, is commonly consumed in different parts of the world and used as a medical drug to treat numerous diseases. OBJECTIVE: To test the toxicity of saffron extract in vivo; to separate different ingredients in saffron extracts; to examine the cytotoxic effect of saffron and its main components on the growth of different human malignant cells in vitro; to evaluate the mutagenic and antimutagenic activities of saffron extract. METHODS: HPLC with photodiode-array detection was used for semi-preparative separation of different ingredients of saffron crude extract. Colony formation assay was used to determinate the cytotoxic activity of saffron extract and its components on human tumor cells in vitro. Mutagenicity and antimutagenicity assays were performed by the Ames method. RESULTS: Saffron is not toxic, non-mutagenic, non-antimutagenic and non-comutagenic. Twelve components were isolated: crocin-1, crocin-2, crocin-3, picrocroein, acid form of picrocrocin, HTCC-diglycosil-kaempferol trans-crocin-4, trans-crocin-2, trans crocin-3, safranal, crocetin and cis-crocin-3. Saffron extract itself and some of its ingredients displayed a dose-dependent inhibitory activity against different types of human malignant cells in vitro. HeLa cells were more susceptible to saffron than other tested cells. CONCLUSIONS: Taken together, our results and literature data indicate that saffron could be used as a potential cancer chemopreventive agent in clinical trials. PMID- 12587419 TI - [HIV glycoprotein 120: possible etiological agent of AIDS-associated dementia]. AB - The AIDS or HIV associated dementia is a cognitive-motor disease, characterized by a strong deficit of several cognitive processes such as attention, memory, sensory perception, motor control among others. The HIV associated dementia affects 30% of adult to 50% of infant HIV positive subjects. Since neurons are not infected by HIV, its principal target in the brain is microglia. The pathophysiology of this syndrome, therefore, remains to be disclosed. Several hypothesis have been proposed, one of them suggests that opportunistic infections can affect the brain. Another hypothesis suggests that microglia secretes toxic products as a result of HIV infection and those are the ones causing the damage and finally, the hypothesis, suggesting that the brain is damaged as a result of the insult caused by HIV-derived proteins. In vitro studies suggest that the HIVgp120, a viral surface protein, is highly neurotoxic. For example HIVgp120 increases cytoplasmic Ca+2 by two ways: facilitating glutamate neurotransmission increasing Ca+2 conductance, and activating the IP3 pathway, facilitating Ca+2 release from the smooth endoplasmic reticulum. This Ca+2 in turn, activates several internal signaling pathways such as the MAPK pathway. We use an animal model to test the HIVgp120 effect on neurophysiological signals and behavior as well as several pharmacological approaches to prevent the HIVgp120 neurotoxic effects. This review updates with the most recent literature discussing the potential mechanisms implicated in the pathophysiology of the AIDS dementia complex. We, in addition, hope the reader will be able to correlate the clinical symptoms observed in the HIV infected subjects and the HIVgp120-induced behavioral changes observed in animal models. Likewise, we discuss the new drugs we are testing, in order to offer a new pharmacological treatment to the patient. PMID- 12587420 TI - [Obesity and asthma]. AB - Obesity and asthma are two disorders of high and increasing worldwide prevalence. A consistent association between obesity and asthma has been recently found in case-control, cross-sectional and longitudinal studies. This association is more consistent in women after the puberty. Moreover, an improvement in the severity of asthma has been described after weight reduction in obese patients. A causal association between asthma and obesity may represent an additional respiratory work that could increase the morbidity and medical expenditures. The most striking studies which demonstrate association between obesity and asthma and the possible causal mechanisms are reviewed. PMID- 12587421 TI - [Syncope. A diagnostic trap]. PMID- 12587422 TI - [The effect of genetic factors on the pathogenesis of the dyslipidemias following kidney transplantation]. PMID- 12587424 TI - [Surgery as a therapeutic tool]. PMID- 12587423 TI - [The concept of illness and kidney diseases in Nahuatl medicine. Synthesis of Mesoamerican pre-Columbian medicine]. AB - Medicine in Mesoamerican cultures began in the year 1,500 BC and ended with the conquest and destruction of Mexico-Tenochtitlan in 1521 by the Spanish conquerors. Mesoamerica began with the Olmec civilization followed by the Teotihuacans, Toltecs and Mayans, and perished during the Nahoa empire. The medicine used by the Aztecs (ticiotl) is undoubtedly the sum of all Mesoamerican medicine. The medical history of the ticiotl was recovered in the years that followed the conquest from the works of Bernardino de Sahagun, Francisco Hernandez and the Cruz-Badiano codex. All these works described the use of plants and herbs in the treatment of diseases, including edema, urinary retention, kidney stones, and podagra. The Aztec doctors (Titici) were well acquainted with innumerable diseases and were excellent healers of wounds and fractures. The works of modern historians confirm the theory of the ticiotl medicine. The later used a complex and philosophically elaborated medical theory based on, religion, astronomy, divination and the polarity cold/warm; different from the four humor theory of galenic medicine. They demonstrated that every culture is capable of understanding and "inventing" the meaning of disease and its cure, even when it is different from our modern medical views. PMID- 12587425 TI - [Memory of Professor Apolinary Z. Szewczuk 1928-2002]. PMID- 12587426 TI - [Biological activity of synthetic analogs of laminin active sequences: YIGSR, RGD, SIKVAV]. AB - The invasion of malignant cells through the basement membrane is a critical step in local infiltration and metastasis. Adhesion and invasion of malignant cells may be modulated by their receptor mediated binding to the basement membrane glycoprotein-laminin. Laminin consists of the sequences with anticancer and antimetastatic activity. Its peptide fragment YIGSR is known to inhibit the experimental metastases of several tumors. This sequence and a tripeptide RGD of laminin inhibits both angiogenesis and tumor growth. In contrary, the sequence SIKVAV initiates angiogenesis and tumor progression. Moreover, laminin plays also other functions in human organism. One of them is the influence on platelet aggregation and thrombogenesis. PMID- 12587427 TI - [Fatty acids as a potentially helpful supplement in cancer therapy]. AB - This review presents pro- and anticancer effects of fatty acids in vitro and in vivo. The epidemiological and experimental data indicate that short-chain saturated and long-chain unsaturated omega-3 fatty acids exert protective effects against some cancers. In the contrary, omega-6 fatty acids are rather procancerous and can increase tumour proliferation. The possible mechanisms are still not fully understood and are very variable including: suppression of neoplastic transformation, direct cytotoxicity, apoptosis induction, antiangiogenicity etc. Experimental data indicate that fatty acid composition of cancer cells in cultures or in animals can be modified to make the cells more susceptible to lipid peroxidation, increasing the cell damage due to oxidant stress and make the cells more sensitive to chemotherapy. It is reasonable to suppose that fatty acids supplementation can make an enrichment of traditional tools for modern anticancer strategies. PMID- 12587428 TI - [The role of membrane glycoproteins CD46, CD55 and CD59 in protection of tumor cells against complement lysis]. AB - Utilization of the complement system offers potential for the elimination of tumor cells by monoclonal antibodies (mAb) immunotherapy. Activation of the complement system causes tumor cell destruction by inducing complement lysis and promoting cell-mediated killing. In addition, complement can induce a strong inflammatory response, which might enhance other antitumor effector mechanisms. An important targets for mAb immunotherapy, however, are membrane bound complement regulatory glycoprotein: CD46, CD55 and CD59, which have been found to be expressed on most tumor cells in vivo and in vitro. Blocking or down regulation of these inhibitors could be an important step in the advancement of mAb immunotherapy. PMID- 12587429 TI - [Effect on manganese on the circulatory system]. AB - This review describe the effect of manganese on the heart and blood vessels. The interaction between manganese and redox systems and manganese contribution to atherosclerosis development were also investigated. The results of the experimental studies on animals, on isolated blood vessels in vitro and on people professionally exposed to manganese were presented. PMID- 12587430 TI - [The role of L-ascorbic acid in free radical reactions]. AB - L-ascorbic acid is one of the basic vitamins, necessary to the normal growth and behaviour of organisms. The primary of this review is to present the chemical characteristics of this substance and to discuss its relation to various biological functions of vitamin C, mainly as an antioxidant barrier (we analysed product of reaction between phenothiazine derivative and L-ascorbic acid). PMID- 12587431 TI - [Multilevel regulation of tyrosine hydroxylase. II. Transcriptional regulation]. AB - The tyrosine hydroxylase gene is subject to very precise regulation which aim is to adjust the level of the catecholamines to current stimuli disturbing the homeostasis. The fine tuning of the TH gene activity is realized by the "cross talk" between the complexes of transcriptional factors with their appropriate regulatory sequences. The transcriptional aspect of that regulation has been reviewed emphasizing the rule of the regulatory sequences in determining cell, tissue and developmental specificity of the TH gene activity. PMID- 12587432 TI - [Privatization of the Croatian health care system: effect on indicators of health care accessibility in general medicine]. AB - The aim of the privatization of the primary health care is to reduce cost and improve the quality of service by introducing the market-based system. In the Republic of Croatia, the privatization of the primary health care started in 1995. It was based on renting the existing facilities at a moderate price to the practitioner. After that, the practitioner registers the private practice and signs a contract with the obligatory health insurance institution. The aim of this article is to present a part of the project of long-term research of privatization related to the health care accessibility in general practices. The research sample consisted of three groups: privatized general practices, to-be privatized general practices and non-privatized general practices during 1997 and 1999. Privatized general practices have exhibited a significant improvement in the number of registered patients, the scheduling of first visit appointments during working hours, the possibilities of scheduling follow-up visit appointments during working hours, scheduling visits by telephone, obtaining telephone advice after working hours and visiting the practitioner after working hours. This kind of research is vital in order to bring about the necessary changes in the existing system by scientifically approved methods. PMID- 12587433 TI - [Epidemiologic characteristics of patients with myocardial infarct in the region of Split during the period of war in the homeland]. AB - The aim of this study was to determine the pattern of myocardial infarction (MI) incidence regarding the age, gender, infarction site and the most important risk factors. Between 1989 and 1997 there were 3454 patients hospitalized in coronary care units of Clinical Hospital Split. In the three-year period preceding the war, from 1989-1991, 1024 patients were hospitalized because of MI. During the three years of full was activities, from 1992-1994, there were 1257 patients (significantly more, p < 0.05), and in the three-year period after the was, from 1995-1997, there were 1173 patients. In the war period there were 12% (151) patients under the 45 years of age (p < 0.05); of that number, 95% (143) were men (significantly more than in other two periods, p < 0.05), and 5% (8) were women. In the period preceding the was there were 6.5% (66) patients under the 45 years; 91% (60) men and 9% (6) women, whereas in the period after the war there were 7.5% (88), 92% (81) and 8% (7), respectively. The patients under 45 (305) more often had MI of inferior than anterior site (49 vs. 28%, p < 0.001), whereas there was no difference in patients over 45 (36 vs. 37%, p > 0.05). The patients over 45 had significantly higher hospital mortality (21 vs. 4%, p < 0.001), and were more likely to have hypertension (51 vs. 15%, p < 0.001) as well as hypercholesterolemia (54 vs. 14%, p < 0.001). Smokers were more prevalent among those under the 45 (75 vs. 51%, p < 0.001). The number of hospitalized patients with MI was the greatest during the war period. It included significant increase in incidence in men under 45 (12 vs. 7%, p < 0.05), with smoking as the most important risk factor, especially for infarctions of inferior site. PMID- 12587434 TI - [Respiratory symptoms, diseases and pulmonary ventilatory capacity in persons in the third stage of life]. AB - The study included 622 persons of the third age, 60-94 years old. The subjects were analyzed by age, smoking habits, profession and place of longest residence. In the studied group, there were 27.2% men smokers and 12.6% women smokers. In relation to profession, women were mostly office employees (35.4%) and men were mostly workers (46.9%). Ventilatory capacity was measured by recording maximum expiratory flow-volume on which forced vital capacity (FVC), one second forced expiratory volume (FEV1), and maximum expiratory flow rates at 50% and the last 25% of vital capacity were read (FEF50' FEF25). All respiratory symptoms were significantly more prevalent in smokers than in nonsmokers. The highest prevalence was found for chronic cough in men (31.1%) and in women (30.1%) as well as for dyspnea in women (54.7%) and in men (43.5%). A higher prevalence of respiratory symptoms was found in the age group 60-69 years old than in subjects older than 70 years. Asthma was more prevalent in workers and farmers, which is probably the result of work exposure. Logistic regression analysis of chronic respiratory symptoms and diseases in older men and women by profession, age, place of longest residence and smoking habits did not show statistically significant difference between effects of those variables on the development of respiratory symptoms and disease. The measured values of ventilatory capacity tests were decreased up to 70-79 years of age. After that there is no decrease expressed as a percentage of reference values. PMID- 12587435 TI - [Successful treatment of refractory hypotension during hemodialysis using high levels of sodium in the dialysis solution]. AB - We report a case of grave hemodialysis associated hypotension refractory to common therapy and successfully resolved with extremely high concentration of sodium in dialysate. We briefly discuss pathogenesis and treatment of hemodialysis associated hypotension, and its prevention with modern hemodialysis machines. PMID- 12587437 TI - [Catheter placement for peritoneal dialysis using videoendoscopy]. AB - From December 1993 to December 2000 at the Department of Surgery, General Hospital "Sveti Duh", and University, Hospitals Split and Osijek the catheter for peritoneal dialysis was placed in 128 patients, in 137 cases. Videoendoscopic procedure, with the trocar made precisely for that purpose, was used. All surgical procedures, performed in the completely equipped operation room and in general anesthesia, were done without any intraoperative or postoperative complications. The procedure showed to have all advantages of endoscopic or minimally invasive surgery. The patients recovered quickly with low consumption of analgesics, got out of the bed and started with feeding sooner. There were no wound complications. It was possible to start with the dialysis 2-3 days after the procedure. Most important, the results showed that using this approach in comparison to others, there were no more dialysis solution leakages, catheter drainage problems, or more tunnel, exit site or peritoneal infections. Along with placing the catheter, other surgical procedures could be also done without increasing the complication rate. Therefore, laparoscopic cholecystectomy was done in 12 cases, adhesiolysis in 25 cases, and right adnexectomy, open hernioplasty and umbilical hernioplasty in one case. Videoendoscopic approach of placing the catheter is a simple, short and patient convenient procedure, with quick recovery and without any increase in complication rate. PMID- 12587436 TI - [X-linked adrenoleukodystrophy--case reports of 4 patients in a family]. AB - A result of a thorough clinical and laboratory study of 27 family members from three generations showed three patients with adrenoleukodystrophy (ALD), one with adrenomyeloneuropathy (AMN) and five females heterozygous for ALD, three of which were psychiatric patients. Four males died at younger age under mysterious circumstances and it is certain that three of them had dark pigment. Based on this information, and on their position in the family tree, it can be presumed that all of them, or three at least, had ALD or AMN. It is necessary to measure very long chain fatty acids level (VLCFA) in the blood of males with Addison's disease in families with ALD or AMN cases, as well as in persons showing signs of demyelinization of white substance followed by progressive neurological symptomatology of unknown cause. All the ALD and AMN patients detected up to now were diagnosed at the Department of Pediatrics, University Hospital Rebro Zagreb. It undoubtedly indicated that there are still a significant number of undetected cases among children and adults in Croatia. In view of the recently provided possibilities of VLCFA level measurements in Croatia, a larger number of detected cases can be expected. Early detection of patients and heterozygotes for ALD, as well as the prenatal diagnostics, enable the families at risk to plan their descendants. Adrenal insufficiency in these patients is very successfully cured with gluco and mineralcorticoid substitution therapy. Unfortunately, there are still no methods of stopping or curing progressive neurological disorders. PMID- 12587438 TI - [Venous thrombosis at the beginning of the new millennium]. AB - The author presents new theories related to the etiopathogenesis, diagnostic procedures and therapy of the deep vein thrombosis, according to latest publication data. Dilemmas related to the diagnosis and therapeutic strategies in hospital as well as the possibilities of treatment on the outpatient basis will be discussed. The intention was to cut the costs of therapy in favour of low molecular weight heparin treatment on the outpatient basis, as the cornerstone of the new therapeutic possibilities besides the importance of fibrinolytic therapy. Hereditary and acquired thrombophilia and venous thrombosis related to pregnancy will be discussed. PMID- 12587439 TI - [BOOP-bronchiolitis obliterans organizing pneumonia]. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) is increasingly recognized in "Jordanovac" Clinical hospital for lung diseases. The characteristic pathohistologic finding is the granulated tissue plugs within the lumen of small airways which extend into the alveolar ducts and alveoli. Numerous conditions and diseases show such histologic picture but the clinician by means of clinical, radiologic and laboratory findings establishes the final diagnosis. By eliminating the known causes and conditions of this syndrome the diagnosis of idiopathic BOOP is established. Idiopathic BOOP is unequivocal clinicopathologic entity which is in typical cases recognized as a pulmonary infiltrate accompanied by febrile illness of a few weeks' duration that is not responsive to a typical course of antibiotics. Corticosteroid therapy causes the complete disappearance of pulmonary infiltrates in 65% to 85% of cases but relapses are common. The therapy of secondary BOOP is less efficient. The aim of this review is to acquaint the readers with the BOOP so it could be incorporated in the differential diagnosis of febrile noninfective diseases that resemble pneumonia. PMID- 12587440 TI - [Medical science during the Nazi period: on the lecture by Daniel C. Angetter "Treatment and medical experimentation in the Dachau concentration camp"]. PMID- 12587441 TI - [Bacterial resistance to antibiotics in Croatia in 2001]. PMID- 12587442 TI - [Surgical anatomy of the spleen with special emphasis on its segmental architecture]. AB - The authors have analyzed several aspects of the surgical anatomy of spleen, commencing with historical data, topography, peritoneal ligaments, variations in shape, embryology and accessory spleens and venous system of the spleen. The mode of splenic artery branching, variations of polar arteries, and intra- and extraparenchymatous arterial anastomoses were thoroughly analyzed. It was shown that the spleen in most cases consists of five vascular territories (segments) clearly demarcated from each other, stressing the practical significance of splenic anatomy in segmental dearterialization of the spleen. PMID- 12587443 TI - [Elective resection of the spleen--overview of resection technics and description of a new technic based on radiofrequency coagulation and dessication]. AB - The authors present a short overview of the development of elective splenic resections. Past and present indications are presented. Contemporary hemostatic technique for elective splenic resection are discussed. An original new technique for transsegmental partial splenic resection using RF generator Radionic Cool Tip(without any aditional hemostatic procedures is presented. This technique is inovative and when use properly it is a practically zero blood loos technique. A patient with transsegmental splenic resection using RF generator is presented. Further clinical application of the technique is necessary. PMID- 12587444 TI - [Preparation of patients with hematologic diseases for splenectomy]. AB - Splenectomy in haematological patients is usually performed as an elective surgery. These patients may have thrombocytopenia or coagulation abnormalities which have to be corrected as much as possible before surgery. The increase of number of platelets may be achieved by increase of dose of steroids, with immunoglobulins, with danazol or by transfusion of platelets. The coagulation abnormalities are to be corrected by appropriate treatment. The risk of postsplenectomy sepsis may be reduced by proper preoperative vaccination and/or by antibiotics. PMID- 12587445 TI - [Late results of splenectomy in patients with chronic immune thrombocytopenic purpura]. AB - Splenectomy is definitive treatment for idiopathic thrombocytopenic purpura (ITP) because it removes both the sites of autoantibody producing cells and also the major site of platelet destruction. The purpose of this study was to evaluate long term results of splenectomised patients with ITP and to determine predictor factors for good response. A 167 patients with chronic ITP (136 females, 31 males), median aged 35 years (17-74) was splenectomised after 2 to 160 months (Median 12) from diagnosis of ITP. Indications for splenectomy were: 6 weeks of steroid therapy with platelet count below 10 x 10(9)/l or 3 months with platelet count under 30 x 10(9)/l, or treatment with prednisone above 30 mg more of 6 months to increase platelet count over 30 x 10(9)/l, or repeated relapses. Postoperative complications developed in 16 patients (9.5%), 3 of them died (1.8%) due to thromboembolism and 17 patients discontinued later controls. During follow up to 172 months (Median 62) 111/147 splenectomised patients were in remission (75.5%), 99 in complete (above 100 x 10(9)/l), 12 in partial (50-100 x 10(9)/l) and 36 patients (24.5%) were relapsed (below 50 x 10(9)/l). Remission was achieved in 79/88 patients (89.8%) with good response to prednisone before splenectomy toward 32/62 patients (51.6%) with poor response to prednisone (p < 0.01). Remission was obtained in 9/11 patients (81.8%) who responded well to intravenous immune globulin (0.4 g/kg x 5 d) and only in 1/8 who did not (p < 0.05). Higher response rate was achieved in patients under 40 years of age (81.6%) than in older ones (63.4%) (p < 0.05). No difference was shown between sex and time intervals (3, 6, 12, 24, 36 or over 36 months) from diagnosis to splenectomy. Splenectomy is an effective treatment of refractory ITP with response rate of 75.5% after median follow up of 62 months. In our patients better results on splenectomy were associated with age less than 40 years, good responses to steroid, and intravenous immune globulin. PMID- 12587446 TI - [Importance of splenectomy in the treatment of patients with hairy cell leukemia]. PMID- 12587447 TI - [Recurrence of immunologic thrombocytopenic purpura due to hypertrophy of retained accessory spleens]. AB - Idiopathic thrombocytopenic purpura is an autoimmune disease in which macrophages of reticuloendothelial system, mainly in the spleen, remove platelets covered by autoantibodies from circulation. By removing the spleen 60-80% of patients are cured. Partial remission is achieved in 10-20% cases. Very few patients do not react on splenectomy. Recurrency of idiopathic thrombocytopenic purpura in a splenectomized patient after already achieved complete remission, may be caused by hypertrophy of one or more of the retained accessory spleens. We present 3 patients, 41, 23 and 44 year old, in whom splenectomy for ITP had been performed 10, 3 and 11 years earlier. After full remission which lasted 10, 2.5 and 10.5 years a full recurrency of ITP took place with signs of severe thrombocytopenia and haemorrhagic syndrome. Using ultrasonography, computed tomography and scintigraphy accessory spleen/s, were discovered. By their removal, a full remission was achieved in all 3 patients, but full favorable effect appeared approximately three months after surgery during which period additional steroid therapy was necessary. PMID- 12587448 TI - [Splenectomy technic]. AB - Splenectomy has been performed in trauma in a number of benign lesions of the spleen and in a certain haematological diseases. When performed for trauma a luxation technique of splenectomy is applied since it allows quick haemostasis. When performed in other indications, the spleen is usually enlarged. In those patients "splenectomy in situ" is recommended. That means, first, division of the short gastric vessels and gastrosplenic ligament, second, ligation of the splenic artery and finally division of the hilar vessels after which the spleen may be simply removed. Details of operative technique, management of accessory spleens and prevention of operative complications are described. PMID- 12587449 TI - [Cesarean section combined with splenectomy in severely resistant immune thrombocytopenia]. AB - Immune thrombocytopenic purpura (ITP) associated with pregnancy often involves considerable risk both for mother and child, and usually worsens in the third trimester of gestation. Pregnancy and delivery are especially difficult in patients with severe ITP (platelet count below 20 x 10(9)/L), who are resistant to prednisone and high dose intravenous immunoglobulin (IVIgG). In those cases we applied cesarean section (CS), to prevent intracranial haemorrhage due to fetal/neonatal ITP, and splenectomy at the same time as an effective therapeutic strategy for ITP. We present 5 patients (4 with chronic ITP and 1 with ITP associated with HIV infection), aged 21-35 years, with severe ITP (platelet count 2-10 x 109/L), resistant to prednisone (1-2 mg/kg), and 2/3 were resistant to IVIgG (0.4 g/kg x 5 d). Four patients with severe resistant ITP were supported with 1-2 doses of platelets from cell separator before CS and 1-3 dose during splenectomy. One patient increased platelet count to 55 x 109/L after treatment with IVIgG and splenectomy following CS were done without platelet transfusion. Splenectomy was performed immediately after CS in all patients, and two of them were hysterectomised (one with HIV infection). After splenectomy, platelet count was normalised in all patients, and they had no haemorrhage, wound haematoma formation or any adverse events. But ITP relapsed in 2/5 patients after 1-2 months. Two newborns had severe thrombocytopenia, which solved spontaneously after 3 days in one or after treatment with IVIgG in other. We propose that splenectomy following cesarean section should be considered as approach for delivery and treatment option for mothers with severe resistant ITP. PMID- 12587450 TI - [Second hemorrhage in patients with splenic injuries]. AB - Although the diagnosis of spleen injuries is not a considerable clinical problem today, subsequent ruptures of this organ may occur in a smaller number of patients (2-5% of total proportion of spleen injuries) following the so-called "free interval". Such injuries are most commonly explained by present hematoma localized in the central spleen, which becomes larger in time, and eventually causes its rupture. This form of lesion may be found both in isolated blunt abdominal injuries and in associated injuries. When it is the question of delayed hemorrhage, our results as well as data obtained from foreign literature, suggest three basic rise factors of the etiology of this type of injury. These are as follows: a) spleen injuries in severe trauma or polytrauma, b) older patients (over 65 years of age), and c) in cases when more than a single blood unit had to be administered for the initial hemodynamic stabilization of a patient. Delayed hemorrhage, which is occult in polytraumatized patients since it is frequently "disguised" by severity of clinical picture and traumatic shock, may subsequently cause sudden fall of hemogram and hemodynamic parameter values, and if immediate surgery is not performed, it may lead to heavy bleeding and lethal outcome of the patient. PMID- 12587451 TI - [Splenic injuries and mortality in polytrauma]. AB - Severe trauma is the leading cause of death in the working population. Traffic accidents are the most frequent etiological factor, with substantially more male than female victims. In this paper we have analyzed 53 cases of dead patients, among 240 severe traumatized treated in Center of emergency surgery, Emergency Center, CCS from January, 2000-June 2002. Spleen is the most frequent injured abdominal organ. In 34% surviving period was less than 72 hours. One-third of patients required at least two surgeries, including laparotomy. The most preciously diagnosis were performed in the cases of spleen injuries. The most common cause of death was cariopulmonal insufficiency, according both clinical diagnosis and forensic findings. PMID- 12587452 TI - [Partial resection of the spleen in the treatment of benign and traumatic lesions]. AB - It has been thought that the spleen is an organ without important functions, until recently. That is, why splenectomy has been the procedure of choice in a treatment of splenic diseases. Even now, when we know the functional [figure: see text] importance of the spleen, splenectomy is performed frequently, regardless of its complications. The need of spleen functions salvage, favours partial resection of the spleen as competitive in a treatment of its traumatic and benign lesions. Improvement in diagnostic procedures, surgical techniques, transfusiology and postoperative treatment, will promote it as a treatment of choice. The authors of this study have experience with 17 partial resections of the spleen for traumatic, 11 for benign lesions of the spleen, and one ectopic spleen with hypersplenism, without mortality and with insignificant complications. PMID- 12587453 TI - [Surgical treatment of hematologic disorders of the spleen]. AB - Splenectomy--the surgical removal of spleen is being performed in cases of: traumatic spleen rupture, as part of other surgical procedures, number of hematological, infectious and metabolic disorders. During the years 1988.-2001., there were 396 splenectomies performed at the First surgical clinic, for the cause of: autoimmune disorders 187 (47.34%), lymphoproliferative diseases 89 (22.59%). Hodgkin disease 35(8.94%), myeloproliferative disease 39 (9.95%), as part a of "staging" laparotomy 37(9.34%), other hematological disorders 7(2.20%). The spleen of [table: see text] 244 patients weighted 500-1500 g(61.62%), in 56 patients (14.14%) weighted less than 500 g, and in 96 patients (24.24%) spleen weighted more than 1500 g. Patients with thrombocytes less than 40,000/l 16 (4.04%) were perioperativly treated with fresh thrombocytes. Postoperative morbidity and mortality were registered in 54 (13.64%), i.e. 8 (2.02%) patients. Delayed results depended on primary disorder, comorbidities and supportive therapy. In this article, the particularities of the operative procedure were discussed, as well as importance of cooperation of surgeon and hematologist in perioperative treatment. PMID- 12587454 TI - [Intraoperative and postoperative complications of splenectomy]. AB - Spleen is being surgically removed because of trauma, in diagnostic and-or therapeutical purposes because of the benignant and malignant diseases. The percentage of morbidity during and after splenectomy is relatively low. During surgery might occur bleeding, trauma of the pancreatic tail, stomach, lineal flexure of the colon, left hemidiafragm, left suprarenal gland and upper pole of the left kidney, which must be correspondingly reclaimed during the same intervention. In the early postoperative period, postoperative bleeding, subfrenic abscess, pulmonal atelectasis, bronchopneumonia and left pleural extravasations might occur. Especially is important notification of these events in due time and adequate conservative and surgical treatment. After splenectomy, there is an increase of the number of trombocytes, which might lead to the tromboembolic complications. In the prevention of these complications in the postoperative period prolonged antiagregation therapy is suggested. Postsplenectomy sepsis is very late, general complication of splenectomy, which occurs because of the lower immunity in the child age. To prevent these complications, partial splenectomies, reimplantations of the spleen, prolonged application of the penicillin medicines after splenectomy and antipneumococcal vaccine are performed. PMID- 12587455 TI - [Experimental and clinical possibilities of transplantation of the injured and totally devascularized spleen]. AB - The most severe spleen lesions with conquasation and devascularisation of entire organ, when it is practically impossible to do any preservating surgical procedure, are the true indications for the transplantation of this extremely important immunological organ. We have performed the evaluation of the surgical procedure of heterotopic auto transplantation in the 30 dogs with severe spleen lesions. Simulation of totally devascularized spleen with the lesions of V degree was performed by disrupting all segmental blood vessels with deep and long longitudinal transhilar incision. During the 3 months follow-up period, animals were subjected to numerous explorations in order to macroscopically and histologically valuate the implant. In most cases (80-85%) implants had complete vitality with the preservation of normal tissue architecture, while 15-20% of implants had partial or total fibrosis. There were no mortality and no complications after this preservation procedure. The presence of fibrosis in some implants suggests that the implant preparation should be better performed and that transplantation of larger tissue volume is needed. Enriched with this experimental experience we have performed heterotopic auto transplantation in 2 patients with spleen lesion of V degree (car accident and injury at work) with very satisfactory results. PMID- 12587456 TI - [Partial resection of the spleen and spleno-renal shunt in the treatment of portal hypertension with splenomegaly and hypersplenism]. AB - Some of serious hepatic diseases with cirrhosis may be complicated by portal hypertension, splenomegaly and hypersplenism. Splenomegaly inhibits regenerative processes of the liver, and also intensifies sequestration of the cellular components of blood up to hypersplenism. Cytopenia caused by hypersplenism is aggravated by negative hepatic influence on bone marrow activity-hemathopoesis, and also by recurrent bleeding from oesophageal varices, and from the other site of gastrointestinal tract. This circle of pathologic conditions may be interrupted only by liver transplantation, until which patients are jeopardized by acute bleeding and chronic anemia. Partial resection of the spleen and splenorenal shunt may correct portal hypertension and hypersplenism, prevent gastrointestinal bleeding, and alleviate hepatic regenerative processes inhibition. In this study, 51 patients with partial resection of the spleen and splenorenal shunt, were analyzed. PMID- 12587457 TI - [Autotransplantation of the spleen]. AB - Auto transplantation of the spleen can be performed in the patients with traumatic rupture of the spleen, in whom spleen could not be conserved in the other way. The right indication for this method is isolated rupture of the spleen (concvasation or complete devascularisation). This method is not recommended in the endangered patients, patients with previous disease of the spleen as well as in the patients with the perforation of the other abdominal organs at the same time. Auto transplantation was performed in 12 patients with isolated splenic rupture and hematoperitoneum, 11 men and one woman. The majority of patients are younger. In 8 patients, autotransplant was placed into big omentum, in three into lipomatous tissue surrounding left kidney, and in one into anterior abdominal wall. In all the patients from this group, following analysis were taken: MCV (middle volume of erythrocytes), HTC, Hb, Le, Glucose, urea, creatinin, sodium, potassium, alkali phosphatasis, target cells, Howell Jolly's bodies, Heinz's bodies, IgG, IgA, IgM, C3, C4, T3, T4, T8, B, segmentated, eosinophiles, lymphocytes, reticulocytes, thrombocytes, fibrinogen, PT, APTT, aggregation of thrombocytes and aggregation of thrombocytes on collagen. The same parameters were taken in 12 patients with surgery similar to splenectomy and in 12 after splenectomy. After splenectomy, there was decrease of the immunologic defending abilities of the organism because of the loss of the childrens function of the spleen, decreased level of the opsonines and tutsin, which leads to the impaired phagocytosis, decreased concentration of IgM and T and B lymphocytes, while in patients after auto transplantation the results were physiological. The most important thing in the assessment of the function of the autotransplanted spleen is scintigraphic investigation using 99mTc-denaturated red blood cells. In our study, auto transplant function was assessed in 10/12 patients by scintigraphy. Five years after surgery, no one patient was proved to have postsplenectomic sepsis. PMID- 12587458 TI - [Laparoscopic splenectomy--indications, specifics of surgical procedures and surgical technics]. AB - The surgical management of splenic disorders has changed considerably over past few years. The increased use of laparoscopic approach for general surgical problems has prompted surgeons to investigate feasibility of laparoscopic splenectomy. However, some unique anatomical features of disordered spleen may limit the application of laparoscopic techniques for this procedure. In this article we analyzed indications for laparoscopic a surgery of the spleen, some surgical particularities of laparoscopic splenectomy, as well as the techniques to accomplish this procedure. PMID- 12587459 TI - Sphincter and nerve preserving total mesorectal excision. AB - The summer of 2002 has seen the introduction of two new key words in the literature on rectal carcinoma. The words are TME Workshop. Papers attesting to a major impact on rectal cancer outcomes in Norway, Sweden, and Holland, are the background to two major papers in the August British Journal of Surgery, one from Ireland and one from Sweden plus a leading article from a professor in Singapore entitled "Adjuvant therapy for rectal cancer cannot be based on the results of other surgeons". PMID- 12587460 TI - What role, if any, for laparoscopic surgery in Crohn's disease of the hindgut? AB - An outsider to the field of surgery would probably take it for granted that surgeons have a highly developed rationale for choosing a laparoscopic approach to Crohns disease. After all, an increasing number of surgeons are performing laparoscopic surgery for Crohns disease as witnessed by several articles published in the 1990s (Table). In fact this is not quite true. Most papers are case reports or series without controls, capable only of suggesting feasibility. Furthermore, comparison studies often feature selection flaws, and therefore beg the question of whether laparoscopic surgery should or not be considered as standard care. An attempt is made herein to give readers a concise insight of the evidence available in the English language literature. It does not pretend to offer a comprehensive review of the topic rather, it highlights some relevant issues, and then outlines what role, if any, laparoscopic surgery should play in Crohn's disease. There are at least 6 categories for discussion. PMID- 12587461 TI - The role of surgery for bowel incontinence. AB - It is now recognised that the results of surgical treatment for bowel incontinence are frequently rather disappointing. It is wrong to raise patients expectations and push patients into surgical treatment for bowel incontinence unless they are aware of the likely outcome measures. Frequently there is deterioration in continence with age, often surgical treatments last only a short period. PMID- 12587462 TI - Adjuvant treatment of colorectal cancer. AB - Colorectal cancer is a leading cause of morbidity and mortality, with approximately 300,000 new cases and 200,000 related deaths in Europe and the USA each year. Adjuvant treatment of colorectal cancer is now widely accepted and can reduce mortality with approximately 10%. This can be considered as one of the major achievements in oncology from the past decade. Current results will be discussed and strategies for the future will be outlined, including on-going or planned large-scale trials with new drugs and approaches. PMID- 12587463 TI - Local recurrence and five year survival after abdominoperineal resection of the rectum due to rectal carcinoma. AB - In the period 01.01.1991-12.31.1996, 523 operations due to rectal carcinoma were performed on the First Surgical Clinic, the Third Department for Colorectal Surgery. Most common localization of tumor was in the distal third of the rectum 65.2%. In the middle third, there were 28.9% and in the upper, intraperitoneal third 5.9%. We performed 286 low anterior stapled resections, 93 anterior resections with hand-sewn anastomosis and 144 Abdominoperineal excisions of rectum (Miles procedure). Pathohistological examination revealed adenocarcinoma in all cases. In this study we analyzed local recurrence and five-year survival after long-term follow-up in the group where Miles procedure was carried out as a potentially curative procedure (except 4.9% cased with Dukes D stage). There were 74.3% males and 23.7% females median age 59.2 years. According to Dukes classification there were 4.9% in stage A, 47.2% in stage B, 43.1% stage C, and 4.9% stage D. There were 4(2.7%) postoperative deaths. Recurrence of the disease was registered in 44 (30.5%) patients. Local recurrence alone was found in 14 (9.7%) patients, while distant spread was registered in 30 (20.8%) patients. At present, the median follow-up is at 72.9 months. Analysis by the Kaplan-Meier's test shows cumulative survival of 61%, and disease free survival of 63.4% at 60 months of the follow-up. Dukes C is associated with a very poor prognosis; survival after 60 months of follow up shows cumulative Survival of 0.35 while Dukes B has far better prognosis (0.86). Analysis of disease free survival by Dukes stage shows that Dukes C has the worst prognosis (disease free survival 0.36 after 60 months), while stage B has much better prognosis (0.84). Local recurrence analysis by the Kaplan-Meier's test shows disease free survival of 84.9% at 60 months of follow-up. Analysis of local recurrence by Dukes stage shows 1.00% disease free survival for cases in stage A, 0.94 for Dukes B and 0.66 for Dukes C, while overall comparison between groups regarding local recurrence using the Wilcoxon (Gehan) statistic shows statistically significant difference (p-0.005). There is no statistical difference between Dukes A and Dukes B cases in distribution of local recurrence. PMID- 12587464 TI - Modern concepts for the treatment of fecal incontinence. AB - Anal incontinence is estimated to be present in approximately 2% of the total population. However, the incidence of this disorder increases with age, affecting up to 11% of men and 26% of women after the age of 50 years. In general, the causes of long-standing fecal incontinence may be divided into anorectal or congenital malformations, perineal trauma (due to surgery or accident), pudendal nerve lesions with or without muscular injury, and low-motor neuron lesions. Classical surgical treatment includes direct repair of the circumscribed gap in the anal sphincter, the so-called overlapping sphincteroplasty or anal repair. In the short term, this method was shown to be very effective in improving continence. Surgical repair of a diffuse weakness of the pelvic floor by application of the postanal repair method has led to more controversial results. While the short-term results are frequently beneficial, full continence is rarely achieved in the long run, especially in patients with imparied pudendal nerve function. All currently used surgical methods focus on the direct mechanical approach to the pelvic floor muscles and/or the anal sphincter. Therefore, the response to this kind of therapy is limited by the presence of a simultaneously existing neurogenic lesion (pudendal nerve damage), as well as by the magnitude and intensity of muscular injury. These problems have been addressed by the development of new methods that focus on the replacement of large muscular defects (Dynamic Graciloplasty) and the treatment of neurogenic causes of fecal incontinence (Sacral Nerve Stimulation-SNS). PMID- 12587465 TI - Preserving the superior rectal artery in laparoscopic [correction of laparoscopis] anterior resection for complete rectal prolapse. AB - Anterior resection for the treatment of full thickness rectal prolapse has been around for over four decades. 1 However, its use has been limited due to fear of anastomotic leakage and related morbidity. It has been shown that high anterior resection is preferable to its low counterpart as the latter increases complication rates. 2 Although sparing the inferior mesenteric artery in sigmoid resection for diverticular disease has been shown to decrease leak rates in a randomized setting, 3 vascular division is current practice. We shall challenged this current practice of dividing the mesorectum in anterior resection for complete rectal prolapse developing a technique that allows the preservation of the superior rectal artery. PMID- 12587466 TI - Sacral magnetic stimulation in puborectalis paradoxical syndrome. AB - BACKGROUND AND PURPOSE: Our earlier studies have demonstrated that sacral magnetic stimulation (MS) in the canine model, in healthy volunteers and in constipated subjects effected rectal pressure rise, decline of the rectal neck (anal canal) pressure as well as rectal evacuation. Based on these results, we studied the effect of sacral MS on defecation in patients with puborectalis paradoxical syndrome (PPS). METHODS: Eleven subjects (8 women, 3 men; age 36-53 years) with PPS were enrolled in the study. The magnetic coil was placed on the back with its center located between L4 and L5. Stimulation parameters were set at 70% of maximum intensity, 40 Hz frequency and 2-second burst length with 2 seconds off. During MS, the rectal neck and gastric (intra-abdominal) pressures were measured. The procedure was performed in the empty and in the full rectum using the balloon expulsion test in the latter. RESULTS: MS of the empty and balloon-filled rectum effected rise of the rectal pressure (p < 0.001), decline of the rectal neck pressure (p > 0.001) and no significant change of the intragastric pressure (p > 0.05). The balloon was expelled to the exterior in all the patients. CONCLUSIONS: Sacral MS succeeded in dispelling to the exterior the water-filled rectal balloon. The method is simple, easy, non-invasive, non radiologic and can be performed on an outpatient basis for the treatment of PPS. PMID- 12587467 TI - Radio (chemo) therapy in locally advanced rectal cancer. AB - Radiotherapy has an role in combined treatment to lower local recurrence in resectable rectal cancer. Radiotherapy also has an established role in nonresectable rectal cancers to increase the operability, but radiochemotherapy is more efficient. Radiotherapy can be administered as a transcutaneous therapy on the megavoltage machines as well as brachytherapy and combined--transcutaneous and brachytherapy. PMID- 12587468 TI - Perspectives and controversies in the treatment of advanced colorectal cancer- from palliation to cure. AB - Advanced colorectal cancer (ACRC) has been traditionally considered resistant to chemotherapy, with no clear benefit in relation to time to progression and overall survival. Nevertheless, nowadays first and second line therapeutic approaches have dramatically changed the natural history of ACRC, and that means that at present the most important decision in a patient is not to treat or no to treat with chemotherapy, but to choose the best chemotherapy schedule at any time. Following this purpose it has been necessary, not only the emergence of new drugs and schedules, but also the understanding of the clinical efficacy. Actually, to the classic parameters such as, objective response, time to progression, and overall survival, new parameters studying clinical benefit have been added. Among these new parameters we can find the weight loss, symptom-free period, quality of life, pain-free interval, etc. Therefore, the really important aim in these situations is the patient clinical status, being less important the survival or the tumor burden reduction. PMID- 12587469 TI - Risk factors in the recurrence of the colorectal cancer. AB - Traditionally, the clinical outcome of colorectal cancer patients may be predicted by pathological staging by either Dukes staging or the UICC-TNM system. However, some of Dukes stage A (approximately 10% of patients) and Dukes B patients (30-40%) will develop local recurrence or distant metastasis years after receiving standard surgical treatments. Therefore it is important to find some other indicators that can predict for recurrence so that we can screen for high risk early-stage patients who may need preventive chemotherapy or other adjuvant therapy. The aim of this study is determination of risk factor for local recurrence in rectal cancer. In this study there has been used and summarized also research records and publications from different clinical hospitals according to actual international literature. Part of elements connected with patient, tumor and genetic and immunological factors remains independent on curative procedures. However better investigation these factors might affect on therapy, frequency of follow-up examinations, and help to detect recurrence at very early phase. Concomitant treatment factors are able to be moderate by surgeons and therapeutics. Therefore precise definition of risk factors might be helpful in decrease recurrence rate in patients with rectal cancer. PMID- 12587470 TI - Do we still need a permanent colostomy in XXI-st century? AB - The aim of this paper is to answer the question: do we still need a permanent colostomy and present the quality of life of patients operated on for rectal cancer. A comparative analysis of patients after abdomino-perineal resection of the rectum vs. patients after low anterior resection of the rectum with preservation of sphincters is presented. When assessing the quality of life of patients, the following issues were considered: a) function of the whole organism and general condition (physical function); b) psychological well-being (psychological function); c) professional activity, relations with relatives and friends, leisure activities (social function) and d) intimate relations (sexual function). In both groups of patients, both after abdomino-perineal resection of the rectum and after low-anterior resection, a significant deterioration in the quality of life was noted. In spite of a good general health condition in the majority of cases (over 60% in both groups), frequent are irregular stools and diarrhoea. Stomy patients complain about uncontrolled passing of gas and urologic problems, while patients with preserved sphincter complain about constipation. Stomy patients significantly more often suffer depression, loneliness and even despair due to low self-esteem and feeling of unfavourable change in body appearance. This feeling is present more often in younger patients and in women. Rectal cancer may cause social disfunction, like reluctance to resume professional activity after surgery, limitation of social contacts, change of model of rest and leisure activities towards more passive forms and forms which do not require the attendance of many people. These phenomena apply to both groups but are more prominent among stomy patients. As stomy patients are usually older, some of these alterations may be due to age. Sexual dysfunction is significantly more frequent in stomy patients. Age may be one of the causes thereof. These problems are significantly more frequent in males. In some cases of colorectal cancer, the best way of management is colostomy. Further studies are needed to elaborate more clear criteria for optimal management of patients with colorectal cancer. PMID- 12587471 TI - New approaches in ambulatory proctology. AB - Ambulatory proctology is till now very underestimated discipline, which is out of interest of big surgeons. But is is a very important field due to incidence of proctological affections and severe social consequences of their inappropriate diagnosis and treatment. We stress the conservative and semi-invasive treatment of hemorrhoids, anal thrombosis and anal fissures. We also mention the other anal pathologies. PMID- 12587472 TI - Micrometastasis in colorectal cancer. AB - Colorectal cancer is one of the most common malignancies in the word and can be usually cured if diagnosed at an early stage. In the European area the estimated number of new cases of cancer in 1995 were approximately 334.000 and 189.000 died of cancer in that year. Despite recent progress in early detection and surgical therapy, the mortality remained unchanged over the past decades. The major reason for this disturbing discrepancy is that occult dissemination of viable cancer cells can occur at any stage of tumorigenesis. Occult dissemination of the tumor cells in patients with operable cancer may be considered a determinant of subsequent metastasis formation. Several groups have therefore designed immunocytochemical and molecular assays to identify such minimal amounts of residual tumor cells that have successfully invaded secondary organs. The question whether circulating tumor cells represent metastatic dissemination or are merely cancer cells without metastatic potential that have detached from the primary tumor, has been debated for over half a century. PMID- 12587473 TI - Septic complications after low anterior rectal resection--is diverting stoma still justified? AB - A retrospective study was designed to determine the effects of faecal diversion on the rate and severity of clinical anastomotic leaks after low anterior resection. The study explored the complications of stoma closure as well. During the period between 1 January 1995 and 30 July 2000, anterior rectal resection was performed on 249 patients with anastomoses created at a 6-cm or smaller distance to the dentate line. In 74 cases, the anastomosis was protected by loop ileostomy. The indications for creating a stoma were evaluated subjectively, by the operating surgeon. In 64 patients, the ileostoma was closed 3 months later. A 'clinical leak' after anterior resection was defined as an anastomotic insufficiency with clinically relevant consequences. The overall rate of anastomotic leak was 6.4 per cent; it was 5.1% (9/175) without and 9.4% (7/74) with a protective stoma. In 8 out of 9 patients, the anastomotic leak that had occurred without a protective stoma warranted laparatomy and defunctioning colostomy. Lavage and drainage of the peritoneal cavity and the presacral space were necessary in 6 out of these 8 cases- and furthermore, the deranged anastomosis had to be removed in 2 patients. Local management was successful in a single case only. Although relaparotomy entails long-term intensive care, all reoperated patients survived anastomotic leakage. Seven patients with a leak despite a protective did not require laparatomy; transanal drainage was appropriate in all cases. There were no fatalities in this group either. Only one fatal complication from suture leakage occurred after stoma-closure. Abdominal exploration was inevitable in almost all patients with a clinical anastomotic leak and without defunctioning stoma. By contrast, patients with anastomotic insufficiency despite a protective stoma were successfully managed without further intra-abdominal intervention. As shown by these results, faecal diversion undoubtedly mitigates the clinical consequences of anastomotic leaks, but cannot prevent its occurrence. When considering the cumulative risk of surgical complications associated with anterior resection, the complications of stoma closure must also be taken into account. Our data confirm that a defunctioning stoma is beneficial for high-risk patients, who are unfit for a second abdominal procedure required to control suture leakage. PMID- 12587474 TI - New approaches to the treatment of anal fissure. AB - According to Antropoli, pathologies of the anal canal are extremely common. About 30 to 40 percent of the population suffers from proctologic pathologies at least once in their lives. In most cases they are more annoying than dangerous. Anal fissure (AF) was recognized as a clinical entity in 1934. It is a longitudinal defect of the anal canal mucosa and anoderm extending usually from the dentate line to the external verge of the anal canal. This defect exposes the lower half or even most of the fibres of internal anal sphincter. AF is almost always accompanied by extensive tension of this muscle. Anal fissures affect all age groups but predominantly occur in the 3rd and 4th decades of life (2, 8, 23). Gathright states that fissure disease causes from 6 to 15% of office visits and 10% of operative procedures in a colorectal practice. The etiology of anal fissure has only been partially explained and remains controversial although spasm of the internal anal sphincter has been recognized to play a main role in the pathogenesis of this disease. Recent studies have cast new light on the pathogenesis of anal fissures. PMID- 12587475 TI - Laparoscopic versus conventional colorectal surgery--a comparative trial. AB - Laparoscopic colorectal surgery had a somewhat difficult evolution. It is technically difficult, with a broad field of mobilization requiring multiple careful prot placement, need to deal with major vascular pedicles and the handling of larger surgical specimens. Aim of the present study is to make a comparison between laparoscopic and conventional colorectal surgery of cancer. PMID- 12587476 TI - [Carcinoma of the anal canal--results of radical radiotherapy]. AB - From February 1995 to August 2001, 34 pts with squamous cell carcinoma of the anal canal (stage T2-35.3%, stage T3-44.1%, stage T4-20.6%, stage N1-29.4%) were treated prospectively by definitive radiotherapy only, delivered with megavoltage linear accelerator, combined with intracavitary brachytherapy in some patients, at the Institute for Oncology and Radiology of Serbia. Total tumor dose was range 55-75 Gy. Median follow-up time was 24.7 mounts (range 6-66 mounts). Overall survival and disease free survival at 5 years were 65.44% and 68.44%. Local control was 82.3% (28/34 pts). Eight pts had local or distant failure. Early and late complication rates were acceptable. In majority of patients the anal sphincter function was preserved. PMID- 12587477 TI - [Theodor Billroth (1829-1894)]. PMID- 12587478 TI - [The role of strictureplasty in the treatment of Crohn's disease]. AB - Crohn's disease is pandigestive disease of unknown aethiology, with tendency to reccurrences. Until now it is impossible to heal this disease either by medical or surgical treatment. All unfavourable consequences of this disease are result of inadequate treatment of complications which are leading to systemic weakening, with further progression of morbid process. Therefore, the principle of timely and minimal invasive surgical procedure has been generally adopted. Introduced in eighties, strictureplasty is a procedure of such characteristics. During the period 1980-2001 this method was used in 1/3 of 126 patients with Crohn's disease. There were 79 strictureplaties performed, 9 long and 70 short. Out of these 42, 12(28.6%) were postoperative recurrences, and there was one case of duodenal strictureplatsty. Postoperative small bowel fistulae were not observed, and there was no mortality in this group. During the follow up period of at least 5 years, in only two cases (4.76%) some functional disturbances of digestive functions were observed, without indications for reoperation. In this article indications and details of operative technique are discussed. PMID- 12587479 TI - Intrathoracic goiter: analysis of 21 cases. AB - Goiters are classified into: the cervical, retrosternal, substernal and intrathoracic. The aim of research is to analysis of intrathoracic goiters (ITG) which include goiters with more than 80% of tissue in the thoracic cavity and intrathoracic thyroid choristomas. METHODS: In prospective non-randomized study were analyzed 21 consecutive patients operated on for intrathoracic non-toxic benign goiters from 1987-98. Fourteen patients with intrathoracic goiters (more than 80% of tissue in the thoracic cavity) and seven patients with intratoracic choristomas were operated. Two groups of ITG were observed according to the expressed symptomatology and surgical approach and complications of operative treatment were compared to the complications of operative treatment of 986 non ITG non-toxic benign goiters. RESULTS: ITG represented 1.1% of the whole number of operatively treated thyroid diseases. One third of patients were asymptomatic. Commonly observed symptoms were dispnea, stridor and dysphagia and there was no significant difference in appearance of these symptoms between the two groups of ITG. In 19% of patients correct preoperative diagnosis wasn't assessed. Cervical approach with sternothomy was used in 11 patients, cervical approach with right thoracotomy in seven, right thoracotomy only in two, and cervical approach only in one patient. Thoracic approach was used in 95% of cases and there was no significant difference between the two groups of ITG. Concerning the operative complications, in two patients transient vocal cord paresis and in one patient postoperative bleeding were verified. There was no statistically significant deference in frequency of operative complications between ITG and non-ITG. CONCLUSIONS: ITG are rare, but might present a difficult diagnostic and complex surgical problem. Common clinical presentation and identical surgical approach in operative treatment justify the common review of the two groups of intrathoracic goiters. With adequate and timely performed surgical approach, in specialized institutions, frequency of complications in operative treatment of ITG is not higher if compared to operative treatment of non-ITG. PMID- 12587481 TI - [Traumatic arteriovenous fistulas]. PMID- 12587480 TI - [Role of the Swan-Ganz pulmonary artery (PA) catheter in evaluation of hemodynamic changes during myocardial revascularization in patients with low left ventricular function]. AB - Coronary artery bypass surgery in patient with bad left ventricular function is a challenge for surgical time. Specially important is monitoring of haemodynamics. We performed this open, prospective, randomized study with the aim to assess haemodynamics and oxygen profile monitoring. 34 pts for coronary surgery (EF < 40%) were divided in two groups. Group A, 17 pts. Received glucose-insulin potassium (GIK) solution. Group B, 127 pts. Received Ringer solution. Haemodynamic and oxygen metabolism parameters were measured in four time points. I after the induction in anesthesia; II after the operation; III 6 hours post op.; IV 24 hour post op. Data are expressed as mean +/- SD, Anova for repeated measures followed by Newman-Keuls testing were used. In both groups were evident deterioration of cardiac function during first 6 h as well as VO2 and DO2, more prominent in Group B. Significant recovery and improvement of cardiac function were evident in Group A after 24 h. CI during the time in Group A improves significantly (2.14 +/- 0.36 v. 3.05 +/- 0.55; p = 0.0002) and difference during the time between groups was p = 0.005. LVSWI improved significantly during the time him Group A (AIII vs. AIV) p = 0.007. Simultaneously. VO2 improves significantly in Group A (103 +/- 21 vs. 164 +/- 30, p = 0.00001) while difference between groups in DO was p = 0.037. Importance of oxygen metabolism monitoring for both, left ventricular function and haemodynamics assessment was evident in our study. PMID- 12587482 TI - [Use of fibrin glue in the prevention of leakage in pancreatico-jejunal anastomoses]. AB - The aim of this study was to test the protective effects of fibrin sealing on the pancreatico-jejunostomy (PJA), the high-risk anastomosis following pancreas head resection. Experimental study was performed on the mongrel dogs, divided in two groups (20 animals each): Experimental group-with end to end "dunking" PJA, protected by temporary occlusion of the pancreatic duct with fibrin sealant/Tissucol/Immuno Ag/, while control group was without any protective procedure. The animals were followed 5 months in order to study: protective effects of such procedure on the PJA quantified with the percent of anastomotic leakage, effects of the exocrine secretion and effects the endocrine function Results: PJA leakage occurred in 13.33% in control group. No leakage was registered in experimental group. Biochemical, histological and electron microscopic study showed slight transitory elevation of amylase levels. Fibrin glue plug was dissolved and pancreatic juice output was reestablished 12th days postoperatively. Long term follow-up showed no damages of the endocrine and exocrine pancreas. Pancreatic duct occlusion with fibrin glue appeared to be an useful method in the prevention of pancreatico-jejunostomy leakage, without negative effects on the exocrine and endocrine pancreas. PMID- 12587483 TI - [Anesthesia in surgery in patients with diabetes mellitus]. PMID- 12587484 TI - [Surgical treatment of lesions of the lacrimal caruncle]. AB - Each lesion which produces a prolong change of the volume of the caruncle necessitates surgical treatment, removal of the caruncle. Caruncular lesions are uncommon. During 15 years on the Institute for eye diseases in Belgrade authors have treated 121 caruncular lesions. Here they present their way of surgical treatment of the caruncle and point out some problems connect with this procedure. PMID- 12587485 TI - [Acute hemolytic transfusion reaction]. AB - Alloogenous blood and/or corresponding haemoproduct transfusion is an efficient and relatively safe supportive treatment. Despite the fact that pre transfusion investigation of both patients and donors ensure high degree of safety of this type of treatment, occurrence of adverse haemotherapy effects is possible and often unpredictable. Acute haemolytic transfusion reaction occur as a consequence of immune conflict between red blood cell membrane agents and specific antibodies present in plasma. Since it is impossible to completely avoid the occurrence of transfusion reactions, wherein acute transfusion haemolytic reaction present a serious, possibly life threatening complication, it is an imperative to continue to improve the knowledge on pathogenesis mechanisms leading to complications associated with these reaction and to define the most efficient therapeutical modalities. PMID- 12587486 TI - [Aspects of anesthesiology in the treatment of the TRALI (transfusion-related acute lung injury) syndrome]. AB - Definition--signs and symptoms which include dispnea, hypertension, high temperature and high productive tracheobronchial secretion. Physical findings are lung oedema in first four hours. Such patients usually require respiratory help. After adequate therapy, symptoms disappeared in 96 hours. In the beginning. TRALI used to be a part of ARDS and it were treated that way. Today, TRALI is understand like substantive group of symptoms. PMID- 12587488 TI - [Combined use of internal mammary and right gastroepiploic arteries in myocardial revascularization with and without extracorporeal circulation]. AB - Use of arterial grafts represent the new approach in coronary artery bypass grafting (CABG) surgery these days. This article represents our experience in use of two or more arterial grafts in combination (internal mammary artery--IMA and right gastroepiploic artery--RGEA). Between March 2000 February 2002, 10 patients underwent CABG with exclusive use of left or both IMAs and RGEA, with or without extracorporal circulation (ECC). In the group without ECC fast truck anesthesia was used. Post CABG catheterization was performed in three patients. There were no 30 day mortality or morbidity. Post CABG catheterization in two patients showed excellent graft patency. One patient continued to have chest pain and after the catheterization we found ostial narrowing of the celiac trunck which was successfully dilated. One of participants had abdominal hernia repair. Our opinion is that use of arterial grafts in CABG surgery has much lower risk, excellent patency and good long term prognosis. PMID- 12587487 TI - [Treatment of severe anemia and gastrointestinal tract hemorrhage in a Jehovah's Witness]. AB - This case report discribes the treatment of 67-year-old Jehovah's Witness with severe anemia and gastrointestinal haemorrhage from gastric leiomyoma and peptic ulcer. Minimal invasive surgery with meticulous hemostasis, controlled hypotension, hyperoxic normovolemia and normotermia were main principles. Minimal blood samples for necessary laboratory parameters and noninvasive onitoring were ways to decrease iatrogenic blood loss. The operative and postoperative period were uneventful and well tolerated. The patient was discharged home after eighteen days and well in follow up period. PMID- 12587489 TI - [Non-functioning tumor of the islets of Langerhans]. AB - About 15% of tumors of Langerhans, islets do not cause any hormone induced syndrome although they sintetise and secrete one or more regulatory peptides. These tumors are most frequently localised in the head and tail of the pancreas. They are usually greater then 5 cm. In diameter and present with pain, jaundice, palpable mass and malaise, rarely with variceal bleeding due to compression of the splenic vein. About 50% of the tumors present with symptoms caused by metastases. We present a 51 year old women in whom during the investigation for paraumbilical pain, predominantly on the left side a tumor of the tail of the pancreas was discovered and subsequently surgically removed. Standard histology showed a neuroendocrine tumor. Immunohistochemistry showed generalised immunoreactivity with antibodies against chromographin A, neuron specific enolasa and glucagon in more then 95% of cells. Somatostatatin was coexpressed in more then 5% of cells, PP in rare scattered cells. No reactivity was found for the other hormone markers. Ten years after surgery the patient has no signs of tumor recurrency. PMID- 12587490 TI - Case report: multidisciplinary treatment of a patient with gas-producing phlegmone. AB - The authors present a 51-year-old patient with a severe case of gas-producing phlegmone following incision of a perianal abscess. Early diagnosis and extensive surgical excision during the first 12 hours from the onset of symptoms are crucial. Treatment of sepsis complicated by multiple organ failure: lung insufficiency (respiratory distress requiring mechanical ventilation), kidney insufficiency (requiring rehydration, furosemid, manitol), circulation (blood derivatives, saline, colloid solutions, cardio tonics, anti-arrhythmic drugs) and liver must be aggressive. Hyperbaric oxygen therapy is essential with repeated identification of aerobic and anaerobic bacteria (hemoculture, tissue sample, wound swab), their sensitivity to antibiotics and repeated surgical debridement of the wound. Following this treatment the patient was transferred to plastic surgery where Thiersh transplants covered skin defects. He survived with an abdominal wall hernia due to a team effort and aggressive multidisciplinary treatment by the general surgeon, anesthesiologist, hyperbaric medicine specialist, microbiologist and plastic surgeon. He refused hernia repair. PMID- 12587491 TI - [News in cardiopulmonary resuscitation in adults]. AB - In the paper it is presented novelties in cardiopulmonary resuscitation of adults. It was indicated to importance of maintenance of cardiac output as main factor of successful resuscitation. In was pointed out defibrilation by automatic external defibrilators (AED), especially of biphasic type. Recapitulation of the nevelties was also stated. PMID- 12587492 TI - Double mucinous cystadenoma of the pancreas associated with thecoma of the ovary. AB - Cystadenomas of the pancreas are rare single and isolated tumors, usually appearing in young and middle aged women. Thecomas are rare usually unilateral, benign, estradiol, much rarer androgens producing ovarian tumors. We present a 19 year old girl in whom we removed two mucinous cystadenomas of the tail of the pancreas, 7 and 2 cm in diameter as well as a thecoma of the right ovary. As far as we know this combination of tumors have not been reported before. PMID- 12587493 TI - Where to incise and/or divide the cystic duct. AB - The study concern was to establish the position of cystic duct incision/division in circumstances of laparoscopic cholecystectomy. Seventy consecutive human cadavers were dissected. Corrosion casting (50) and post-mortem cholangiography (20) were employed. Cystic duct length was 34.6 mm, and in 88.6% cases its length was 1-5 cm. Mean cystic duct diameters next to the gallbladder neck, within the valve and 5 mm proximal to the junction with the common hepatic duct were 1.95, 0.42 and 1.85 mm, respectively. Lateral cysto-hepatic junction was identified in 78.6%, spiral in 10% and parallel in 10%. In 90% cases the cysto-hepatic junction is within 4 cm of the hepatic duct junction. One case (1.43%) of cystic duct entering the right hepatic duct was identified. The valve of Heisteri consisted of three spiral turns in 73% of the cases with a range from 0 to 5. In 3/70 specimens the spiral valve did not exist. PMID- 12587494 TI - [Restoration of elbow joint flexion using pectoral muscle transfer in patients with arthrogryposis multiplex congenita. Part I: surgical method, rehabilitation and clinical results]. AB - PURPOSE OF THE STUDY: In patients with type I arthrogryposis multiplex congenita, elbows are fixed in extension. The m. biceps brachii and m. brachialis are usually atrophic. The main objective of arm treatment is to restore flexion in the elbow that would enable the patient to reach the mouth with the hand as well as maintain active extension of the arm for hygienic purposes. This can be achieved by several techniques of muscle transfer. Of these, we selected and modified the method of Clark based on transfer of distal parts of the greater pectoral muscle. The aim of this study was to develop the surgical procedure in detail, to design the course of rehabilitation and to evaluate the outcomes of treatment. MATERIAL AND METHODS: The procedure for optimal transfer of three fifths of the m. pectoralis major was based on an anatomical study. In addition, a unified course of rehabilitation was developed. Between 1996 and 1999, this approach was applied to nine upper limbs in five patients (age range, 4.3 to 9 years). All the patients were evaluated in terms of their clinical state according to selected subjective and objective criteria. RESULTS: The outcomes of transfer of the m. pectoralis major were evaluated as very good and good in six cases. Active movement of the elbow was in the range of 15 degrees to 95 degrees; useful movement ranged from 40 degrees to 70 degrees. The strength of elbow flexion rated 4 or 4+. In three cases, although muscle transfer restricted active elbow extension it did not prevent the upper limb from being used for hygienic purposes. In the remaining three cases, the method failed to restore the patient's ability of reaching the mouth. The analysis of unsuccessful results showed that these were always related to a very limited pre-operative passive flexion of the elbow, restricted movement of the shoulder joint and a failure in distal fixation of the muscle transposed. DISCUSSION: Our results show that transfer of the distal three fifths of the m. pectoralis major, performed by a modified method of Clark, was an effective approach because the working capacity of this muscle was comparable with those of the m. biceps brachii and m. brachialis. Bilateral transfer enabled the patients to reach the mouth with both hands and to use the remaining elbow extension for hygienic purposes. The best function was achieved when active movement of the shoulder had permitted raising the arm above the horizontal and passive elbow flexion had been 90 degrees. Our results are comparable with those reported in the literature. The critical phase of this transfer involved fixation of the m. pectoralis major to the forearm. CONCLUSIONS: The surgical procedure for transfer of the m. pectoralis major and subsequent rehabilitation in patients with type I arthrogryposis are described in detail. Good results are achieved in patients who, pre-operatively, had passive movement of the elbow joint and active movement of the shoulder joint. The method is not suitable for patients with a markedly limited pre-operative movement of both the elbow and the shoulder in whom an alternative surgical treatment should be used. PMID- 12587495 TI - [Radical surgery in tumors of the thoraco-lumbar spine]. AB - PURPOSE OF THE STUDY: In this retrospective study, the outcomes of anterior and posterior approaches, performed either simultaneously or consecutively, in the radical surgical treatment of tumors of the thoracolumbar spine were compared in terms of surgery duration, intra-operative blood loss, neurological findings and complications. MATERIAL: A total of 547 patients with malignant tumors of the spine were treated between 1981 and 2001. Of these, the thoracolumbar spine was affected in 422 cases. Spondylectomy from the combined anterior and posterior approach with decompression, vertebral body replacement and stabilization was indicated in 117 patients, 69 men and 48 women (59% and 41%, respectively). Etiology included metastases in 63 subjects (54%), primary malignant tumor in 37 (32%), benign tumors in 11 (9%) and tumor-like lesions in six patients (5%). Surgery involving two procedures carried out simultaneously by two teams of surgeons was used in 45 cases (38%) and approaches performed consecutively (in either the anteroposterior or the posteroanterior order) were applied in 72 cases (62%). Both groups were nearly identical in relation to the patients' average age and disease etiology. METHODS: In young patients with a solitary tumor of the thoracolumbar spine whose disease had a good prognosis, radical surgery including complete removal of the vertebra affected, decompression of nervous structures, vertebral body replacement and stabilization with 360 degrees fusion was carried out. The simultaneous and consecutive procedures were compared in terms of operative time, intra-operative blood loss, neurological findings and complications. RESULTS: The approaches carried out simultaneously by two teams reduced the total time of surgery and permitted a better correction of the spine affected. This surgical procedure, however, was more demanding in terms of operative skills and involved increased intraoperative blood losses. In 45 patients treated by this procedure, the average operative time was 244 min and intra-operative blood loss was 3313 ml. In 72 patients undergoing consecutive surgery, the average operative time was 345 min and blood loss was 2500 ml. The neurological finding was generally better or unchanged. Of the patients treated consecutively, four (5.5%) experienced deterioration; of those operated on simultaneously, only one patient (2.2%) got worse. Two patients died in each group (2.8% and 4.4%, respectively). DISCUSSION: In order to provide the optimal therapy, each cancer patient should be considered individually with respect to all basic rules of cancer treatment. In this, the radical approach is nowadays preferred. Some authors, however, use only the posterior approach. At our department, the combined anteroposterior approach under one anesthesia is our method of choice with the exception of a serious intra-operative complication such as large blood loss. In this case, the treatment is completed at a subsequent operation one week later. CONCLUSIONS: We prefer an active and radical approach to the therapy of spinal tumors. The simultaneous surgery resulted in an operative time reduced by about 100 min (29%). The consecutive treatment, on the other hand, produced lower blood losses by 813 ml (24%). Complete surgery under one anesthesia was preferred. PMID- 12587496 TI - [Knee joint synovectomy in treatment of juvenile idiopathic arthritis]. AB - PURPOSE OF THE STUDY: The objective of this study was to assess short-term outcomes of knee joint synovectomy in a group of patients with juvenile rheumatoid arthritis and to present the authors' view on this approach. MATERIAL: Between 1990 and 1999, synovectomy of the knee joint was performed in a group of 46 children with juvenile rheumatoid arthritis. This group comprised 19 girls and 27 boys aged 4 to 16 years (mean 9.8 years) with all forms of the disease. A total of 85 synovectomy procedures, including repeat operations, were performed on 58 knee joints. METHODS: Arthroscopic synovectomy was used to treat 21 knee joints, open synovectomy from two approaches was indicated, as a primary procedure, in 37 knees. The assessment of subjective and functional conditions of patients was based on a modified rating systems of Lysholm and X-ray films were evaluated by the Larsen classification. The evaluation was carried out at 1 and 2 years after the primary operation. RESULTS: Articular lesions corresponding to mere synovitis were found in 10 knee joints (17.2%), a developing pannus without erosion was seen in 16 (27.6%) and erosion of the articular surface in 32 (55.2%) knee joints. At 1 year, the value of Lysholm's score rose from 47.9 to 84.3 points and was followed by a decrease to 73.2 at 2 years. Within 2 years of the primary operation, the condition recurred in 9 out of 21 knees (42.8%) treated by arthroscopic synovectomy and in 12 cases (32.4%) operated on by open synovectomy. The relapse was observed mostly in patients with an overall high inflammatory activity and polyarticular and systemic progression of the disease. DISCUSSION: We do not agree with the view of some authors that surgical intervention is not indicated until erosions are radiologically manifested. In children, erosions usually present at a late stage, as shown by 12 findings of articular surface destruction in our group that were not detected by radiography. For indication purposes, we distinguished between preventive and therapeutic synovectomy. We found a significant association between the overall activity and early recurrence of the disease. In 60.7% of the cases (28) with excellent outcomes, this activity was low at the time of surgery. On the other hand, in 75% of the cases (8) with poor outcomes, the activity of disease was very high and had a lasting tendency to recur. These findings are in agreement with the conclusions of several other authors who consider the presence of systemic disease to be a contraindication for synovectomy. Advantages of arthroscopic synovectomy reported in adult patients seem to be relative in children. A good view of and accessibility to individual articular components, which are made an advantage of in the adult knee, are rather exceptional in the "tight" child knee. The evaluation of our patients at a short-term follow-up did not give convincing results although the early effect of synovectomy was very good. The poor outcomes seen in our group, which corresponded with observations of other authors, allow us to learn more about the potentials of synovectomy and thus to promote our policy of a thorough consideration of indications for surgery. CONCLUSION: Indications for surgical treatment in patients with juvenile rheumatoid arthritis are evaluated in cooperation with a rheumatologist after an appropriate conservative therapy administered for at least 6 months. Cases with clear signs of plastic synovitis and skeletal lesions shown by radiography as well as all recurrent conditions are treated by open synovectomy. The state of low disease activity is preferred for surgical intervention. It has to be borne in mind that, from whole range of curative procedures, conservative therapy supervised by a pediatric rheumatologist is the method of choice. PMID- 12587497 TI - [Personal experience with treatment of pseudoarthrosis of the navicular bone of the carpus]. AB - PURPOSE OF THE STUDY: A displaced fracture of the scaphoid bone of the hand may result, according to some authors, in the development of pseudoarthrosis in 10 to 50% of the patients and aseptic necrosis of the proximal bone fragment may develop in 30 to 50% of them. Them surgical treatment of scaphoid bone pseudoarthrosis has recently made great progress and the results achieved by the authors in this field are reported in this paper. These results are discussed in light of the results and opinions presented at the 8th Congress of the World Federation for Hand Surgery held in Istambul in June 2001 and are compared with the achievements made by other authors and published in the most recent relevant literature. MATERIAL AND METHODS: At our department, two standard surgical procedures are available to treat pseudoarthrosis of the scaphoid bone that is free from deforming changes due to arthritis. One approach involves resection of the pseudoarthrosis and subsequent cancellous bone grafting, using osseous tissue from the styloid process of the radius and osteosynthesis with a navicular screw. The other procedure is a modified arthroplasty according to Dwyer and is applied to cases which show aseptic necrosis of the proximal bone fragment. The follow-up study of the patients treated between 1997 and 2001 included patients in whom the follow-up period was at least six months. The results were evaluated according to Pechlaner's rating system. RESULTS: A total of 26 patients were examined; the average time from the operation was 17 months. Of them, 24 underwent the primary treatment that involved resection of pseudoarthrosis followed by cancellous bone grafting and osteosynthesis with a screw. This primary operation healed the condition in 17 patients, but the rest of them had to undergo one or two more surgical procedures. Evaluation by Pechlaner's scoring system showed 14 excellent, five good, two sufficient and three poor outcomes of the treatment. DISCUSSION: The pseudoarthrosis of the scaphoid bone, its diagnosis, surgical treatment and different operative techniques are discussed. Special attention is paid to the use of either vascularized rotatory grafts or pedicled vascularized grafts. The follow-up results are compared with those of other orthopedic clinics and hospitals. Various aspects of further improvement in the treatment of this disease under current conditions are considered. CONCLUSIONS: The results achieved in the treatment of pseudoarthrosis of the scaphoid bone of the hand are comparable with those of other orthopedic hospitals. Adoption of more recent types of osteosyntheses in considered. For a potential introduction of the vascularized graft technique, the use of a vascularized graft collected from the dorsum of the distal radius is suggested. This method is also regarded as suitable for treatment of certain cases of aseptic necrosis of the lunate bone. PMID- 12587498 TI - [Preoperative planning and surgical technic in achieving stability and leg length equality in total hip joint arthroplasty]. AB - One of the prerequisites for a good outcome of total hip arthroplasty is preoperative planning. Using a roentgenogram, the size of an implant, incision level on the femoral neck, depth required for fitting the cup, restoration of the center of hip rotation and, if necessary, correction of length descrepancy between the legs are determined. The preoperative planning based on an X-ray image was introduced by M. E. Muller and, in 1976, was modified by R. Schneider who used a transparent sheet for a template on which all relevant points guiding the surgical procedure are marked, i.e., the right position for implantation of the cup and stem, and incision lines. In uncomplicated cases, however, this approach is not necessary and the "planning principle of parallel lines" developed by L. Spotorno in 1988 can be used instead. The determination of length discrepancy between the legs is derived from a drawing of three reference lines on the roentgenogram. The lines parallel to each other indicace the same length for both legs. If the legs differ in length, the lines will diverge from each other in a way typical for this condition. PMID- 12587499 TI - [Bilateral fractures of the femoral head (Pipkin I and Pipkin II)]. AB - The author describes a rare case of bilateral dislocation of the hip joint accompanied by fracture of the femur head. The fracture (Pipkin I) on the left side was treated conservatively and the fracture (Pipkin II) on the right side surgically. At 18 months of follow-up, both fractures were healed but the fragment of the operated femoral head had a tendency to develop osteoporosis. Aseptic necrosis was absent, as evidenced by bone scan. Both hip joints, however, showed signs of developing arthritis, with clinical findings being less serious on the non-operated joint. The author discuses all cases of bilateral Pipkin fractures reported in the literature and be concluded that there is no report on a coincident occurrence of type I and type II fractures. When comparing conservative and surgical approaches to similar types of fractures, the author recommends the use of conservative therapy. PMID- 12587500 TI - [The Decade of Bones and Joints 2000-2010. Significance of the Decade of Bones and Joints for orthopedics and traumatology]. PMID- 12587501 TI - [11 beta-hydroxysteroid dehydrogenase type 2 activity in Chilean patients with hypertension]. AB - BACKGROUND: Half of hypertensive patients with low plasma renin activity have a primary hyperaldosteronism. Among the remaining half, 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta HSD2) deficiency plays an important role. This enzyme catalyzes the conversion of cortisol to cortisone, avoiding the interaction of cortisol with the mineralocorticoid receptor. If the enzyme fails, cortisol will stimulate sodium and water reabsorption and increase blood pressure. AIM: To determine biochemical alterations, suggestive of 11 beta HSD2 deficiency, in low-renin hypertensive patients. PATIENTS AND METHODS: Twenty eight hypertensive patients with a plasma renin activity of less than 0.5 ng/ml/h and with a plasma aldosterone of less than 5 ng/dl were studied. Twenty eight normotensive patients were studied as controls. Serum cortisol (RIA), cortisone (ELISA) and the serum cortisol/cortisone ratio were determined in all of them, between 9 and 10 AM. Measurements were confirmed by high pressure liquid chromatography. The serum cortisol/cortisone ratio was considered abnormal when its Ln (cortisol/cortisone) value was over 2 standard deviations of the mean. RESULTS: Serum cortisol was higher in hypertensive subjects than in controls (11.1 +/- 3.3 and 9.2 +/- 2.8 micrograms/dl, respectively; p < 0.05). No differences were observed in serum cortisone (3.4 +/- 1.3 and 3.7 +/- 1.2 micrograms/dl, respectively). Four hypertensive subjects had an abnormally high Ln (cortisol/cortisone) value (1.86; 1.73; 2.07 and 2.01, considering a normal value of less than 1.61). CONCLUSIONS: Four of 28 hypertensive subjects with low plasma renin activity and aldosterone had biochemical alterations suggestive of 11 beta HSD2 deficiency. PMID- 12587502 TI - [Cutaneous electrogastrography: a new incorporated technique for the study of gastric motility]. AB - BACKGROUND: Gastric electrical activity can be accurately recorded and analyzed by cutaneous electrogastrography. Different types of abnormalities have been described in a variety of disorders. AIM: To analyze a group of asymptomatic subjects and assess the prevalence and patterns of dysrrhythmias in patients with type 1 and 2 diabetes and patients with functional dyspepsia. PATIENTS AND METHODS: One hundred subjects were studied (32 male, mean age 45 years old, 10 asymptomatic, 11 type 1 diabetics, 22 type 2 diabetics and 57 subjects with functional gastrointestinal disturbances). Gastric myoelectrical activity was recorded using surface electrogastrography for 1 hour in the fasting state and 1 hour after a test meal (350 kcal). RESULTS: The electrogastrogram was normal in 90% of asymptomatic controls. It was abnormal in 64% of type 1 diabetic patients, in 55.5% of type 2 diabetic patients and in 61% of patients with functional dyspepsia. Different types of dysrrhythmias were observed in each group. CONCLUSIONS: Electrogastrography might define a subgroup of patients with electrical rhythm disturbances, that may have a different approach to treatment than patients with normal gastric electrical activity. PMID- 12587503 TI - [Fontan procedure: early and mid-term results with total cavopulmonary anastomosis]. AB - BACKGROUND: During the last five years, 65 patients with univentricular heart have been treated surgically in our institution, according to a protocol of staged operations that have been previously reported. AIM: To evaluate the early and mid-term outcome of those patients that have completed their staging protocol by means of a Fontan procedure. PATIENTS AND METHODS: Between April 1996 and June 2001, 23 patients (age 16 to 223 months) underwent a Fontan procedure, 15 with an intracardiac lateral tunnel technique and 8 with an extracardiac conduit. A retrospective review of their clinical, surgical, echocardiographic, angiographic and hemodynamic data was performed, trying to identify risk factors for both mortality and functional capacity (FC). Follow up was complete in all survivors. RESULTS: Three patients died early after surgery (13.04%). Excessive pulmonary blood flow was a risk factor for early death (p = 0.03). One patient died at 14 months. Follow up was 29.9 months (1-63). For those who survived the operation, five years survival was 93.3%. The majority of patients are in FC I or II, with no related risk factors. CONCLUSIONS: Our current results are comparable with those of larger series. Patients reach good FC and mid-term survival, irrespective of type of single ventricle or the surgical strategy. PMID- 12587504 TI - [Determination of the insulin resistance index by the Homeostasis Model Assessment in a population of Metropolitan Region in Chile]. AB - BACKGROUND: Insulin resistance is defined as an inappropriate high level of plasma insulin required to maintain metabolic homeostasis. It is associated with type 2 diabetes and cardiovascular diseases. The glucose clamp technique is the standard method for the measurement of insulin resistance. However, this method is laborious, expensive and impractical to perform in epidemiological investigations. The homeostasis model assessment (HOMA) has been proposed to assess insulin resistance and secretion, using fasting glucose and insulin concentrations. AIM: To measure insulin resistance using HOMA (HOMAIR) in a population sample from the Metropolitan Region in Chile. MATERIAL AND METHODS: One hundred twenty subjects (59 female) with a normal body mass index and fasting blood glucose were studied. Fasting plasma glucose was measured by a glucose oxidase method and serum insulin was measured by radio immunoassay. RESULTS: Fasting blood glucose was 81.6 +/- 9.4 mg/dl and serum insulin was 9.7 +/- 2.4 microU/ml. Mean HOMA insulin resistance was 1.96 +/- 0.57 (range 0.5 and 3.0). CONCLUSIONS: These HOMA values can be used as reference for Chilean non obese individuals. PMID- 12587505 TI - [Immunohistochemical flow chart to differentiate between primary adenocarcinoma and the most frequent extragynecological metastases in the ovary]. AB - BACKGROUND: The pathological differential diagnosis between primary and metastatic ovarian malignant tumors is usually difficult, specially for tumors originating in the gastrointestinal system or breast. AIM: To define an immunohistochemical flow chart to discriminate these tumor types. MATERIAL AND METHODS: We performed a immunostaining analysis using a panel of six antibodies (CK 7, CA 125, CEA, CK20, BRST-2 and CA 15-3) in 3 tumor groups: 11 ovarian, 14 breast and 12 colonic primary tumors and in a study group of 38 ovarian tumors whose primary origin was unknown. RESULTS: We defined an ovarian immunohistochemical pattern (CEA-/CK20-, 45% in ovarian tumors, 0% in breast or colonic tumors, p < 0.001); an ovary/breast pattern (CEA+/CK20-, 0% in colonic tumors, p < 0.001): a breast pattern (CEA+/CK20-/BKST-2+, 64% in breast tumors and 0% in colonic and ovarian tumors, p < 0.001) and a colonic pattern (CEA+/CK20+/CK7-, 67% in colonic tumors and 0% in breast and ovarian tumors, p < 0.001). CONCLUSIONS: Employing a simple panel of antibodies, characteristic immunohistochemical patterns were defined for ovarian, breast and colonic tumors. These patterns allowed the identification of the origin of most tumors of the study group. PMID- 12587506 TI - [Health care expenditures among the offspring of cocaine base paste consumers]. AB - BACKGROUND: Health care costs of the offspring of mothers addicted to cocaine are three times higher than those of children not exposed to cocaine during gestation. AIM: To calculate the health care costs of the offspring of addict mothers that consumed cocaine during gestation. To verify the diseases or health conditions in these children, that generated the expenses. MATERIAL AND METHODS: One hundred offspring of addict women consuming cocaine base paste were studied. The health care expenses generated by hospital admissions and ambulatory follow up were calculated. Expenses were expressed in Chilean pesos, according to the value at June, 2000. The fares of the South Orient Metropolitan Health Service were used as a reference. RESULTS: Twenty four of the 100 children were lost from follow up. Among the 76 followed children, 48 were admitted to hospitals in 2.3 (range 1.2) occasions. Mean hospital stay was 21.7 days (range 1-186) and hospital mortality was 4%. Mean expense per hospital discharge was $1,556,098 and per patient was $3,457,995. The monthly expenses per children during ambulatory follow up were $120,372 that increased to $395,200 if family placing was added. CONCLUSIONS: These figures confirm that health care expenses of cocaine addicts offspring are more than three times the cost of a normal child. Primary and secondary prevention of cocaine addiction is urgently needed. PMID- 12587507 TI - [Major depression as a new risk factor for coronary heart disease in Chile]. AB - BACKGROUND: Major depression, a frequent psychiatric disease, is associated with ischemic heart disease. It is usually subdiagnosed and subtreated. AIM: To study the prevalence of major depression among survivors of an acute myocardial infarction. PATIENTS AND METHODS: Retrospective study of 42 survivors of an acute myocardial infarction treated at a regional Chilean Hospital. The presence of major depression in the 6 months previous to the acute myocardial infarction, was investigated using the diagnostic instruments CIDI (Composite International Diagnostic Interview) and DIS (Diagnostic Interview Schedule), psychiatric diagnoses were based on DSMIII-R. The prevalence of depression was compared with that observed in a group of 156 healthy subjects participating in a psychiatric epidemiological study. RESULTS: Major depression was diagnosed in 12 male subjects with an acute myocardial infarction. The prevalence in the control group was significantly lower (15%, p < 0.049). Patients with depression were older and required longer hospital stay than patients without depression. CONCLUSIONS: Patients with acute myocardial infarction, had a significantly greater prevalence of major depression in the previous 6 months, than the general population. Thus, major depression could be an independent and modifiable coronary risk factor. PMID- 12587508 TI - [Prevalence of diabetes mellitus in the Seventh Region of Chile]. AB - BACKGROUND: The prevalence of diabetes mellitus in Latin America is not well known. AIM: To study the real prevalence of diabetes mellitus (DM) and its associated risk factors in the VII region of Chile. MATERIAL AND METHODS: A probabilistic sample of 1,325 subjects over the age of 20, stratified by age and differentiated by place of residence was studied. The criteria of the World Panel of Experts convened by the World Health Organization in 1997, was used to define the presence of diabetes (two fasting blood glucose values over 126 mg/dl or a blood glucose over 200 mg/dl, 2 hours after a 75 g carbohydrate oral load). RESULTS: The global prevalence estimated for DM in this population was 5.39%. The calculated prevalence in subjects between 20 and 44 years was 1.88% (CI 0.39 3.37); between 45 and 64 years, 10.75% (CI 8.35-13.14); 65 years or older 11.30% (CI 8.00-14.60), p < 0.05. The prevalence was 5.8% in urban areas and 4.5% in rural areas. Forty five percent of diabetics were not aware of their condition. Forty eight percent of diabetics and 31% of non diabetics were hypertensive (p < 0.01), whereas 47.8% of diabetics and 24.6% of non diabetics had at least one diabetic parent (p < 0.01). Smoking was less frequent in diabetics than in non diabetics (15.7 and 24.3% respectively, p < 0.05). Ninety percent of both diabetics and non diabetics were sedentary. DISCUSSION: Health care systems require an epidemiological monitoring system to provide information about the prevalence of diabetes mellitus and to control the evolution of patients. PMID- 12587509 TI - [Impact of regulatory measures in the trends of community consumption of antibiotics in Chile]. AB - BACKGROUND: The indiscriminate use of antibiotics increases the appearance of bacterial resistance and forces the use of more expensive alternatives. The Chilean Ministry of Health is regulating the consumption of antibiotics since September 1999. These regulatory measures restricted the sale of antibiotics only when these were prescribed by an MD or an DS. AIM: To study the impact of these regulatory measures on antibiotics sales in pharmacies. MATERIAL AND METHODS: A retrospective analysis of antibiotics sales in pharmacies from 1996 to 2000. The information was obtained from the International Marketing System (IMS Health), an auditing system of pharmacy sales. The consumption unit used was the Defined Daily Dose per 1000 inhabitants/day (DDD). RESULTS: There was an important reduction in DDD, after the introduction of regulatory measures, for amoxicillin, ampicillin, erythromycin, trimethoprim/sulpha, chroramphenicol, cloxacillin and phenoxymethylpenicillin. There was also a important fall in sales, expressed in dollars. CONCLUSIONS: The regulatory measures of the Ministry of Health, had an immediate and great impact on antibiotics sales in Chile. PMID- 12587510 TI - [Idiopathic granulomatous hepatitis with bilateral orchiepididymitis and skin eruption]. AB - We report a 52 years old male admitted for fever lasting one month, dry cough, headache and malaise. Initial laboratory work up showed an AST of 172 U/l, and ALT of 252 U/l, a GGT of 353 U/l and alkaline phosphatases of 952 U/l. An abdominal CAT scan disclosed a mild hepatosplenomegaly. A liver biopsy showed a granulomatous hepatitis. During the evolution, the patient had a left testicle swelling with darkening of the surrounding skin. A testicular ultrasound showed a bilateral orchiepidydimitis. The patient was treated with non steroidal anti inflammatory drugs and fever subsided. Three months later, these drugs were discontinued and the patient remained asymptomatic and with normal laboratory values until 36 months of follow up. PMID- 12587511 TI - [Achilles tendinitis associated to levofloxacin: report of 4 cases]. AB - Fluoroquinolone-associated tendinitis are well described in the literature, but these are not frequently observed and usually are related to the oldest fluoroquinolones. Levofloxacin is a recently introduced fluoroquinolone. In this paper we report four cases (three female, aged 39 to 70 years old) of Achilles tendinitis occurring in a period of few months. Two patients on chronic dialysis, one kidney transplant recipient, and one patient with chronic vasculitis, both with normal renal function. All were chronically using corticosteroids. In the four cases, tendinitis had an acute onset with bilateral involvement and was invalidating. In 3 cases the onset of tendinitis was early during levofloxacin treatment and in 1 case, it began 10 days after the end of the treatment. All patients were treated with rest and physiotherapy, two were treated with non steroidal anti-inflammatory drugs. All patients had a complete recovery alter 3-8 weeks. PMID- 12587512 TI - [Mesenteric acute ischemia caused by spontaneous and isolated dissection of the superior mesenteric artery]. AB - Spontaneous dissection of the superior mesenteric artery is an unusual cause of mesenteric ischemia. Diagnosis can be based on findings of the belicoidal computed tomography or selective angiography of the superior mesenteric artery. We report two male patients, aged 47 and 50 years, who were admitted for abdominal pain. Isolated superior mesenteric artery dissection was confirmed by angiography. Both patients were treated with anticoagulation, conservative measures and recovered uneventfully. PMID- 12587513 TI - [Health needs from the perspective of the client]. AB - BACKGROUND: The perception of the population about their needs must be considered in health care planning. AIM: To describe health care needs from the perspective of potential clients of public health services. MATERIAL AND METHODS: An inquiry to a representative sample of 202 dwellings, ascribed to a Family Health Service, was done. Subjects were asked about the health problems in their family and neighborhood. Twelve families from this sample were randomly chosen and interviewed about their health conception, needs and demands. RESULTS: The mean age of subjects, per dwelling, was 31 years. Fifty percent were nuclear families and 6% were mono parental. Perceived problems were economical in 60% and health related in 12%. Eighty percent of health problems were related to addictions and violence. Interviewed subjects associated health with lack of services, with their experience with illness and pain and with a risk of social isolation and incommunication. Health care demands were referred to a better medical care. People declared needs in environmental hygiene, physical and mental health self care, food and jobs. CONCLUSIONS: Using this information, simple strategies could be implemented to cope with health care demands of the population. PMID- 12587514 TI - [Is gallbladder cancer a disease with bad prognosis?]. AB - Gallbladder cancer is frequent in Chile, but it is not uniformly mortal. The diagnosis is usually made after a cholecystectomy, indicated for a symptomatic cholelithiasis. Global survival of gallbladder cancer can be as high as 40% at five years. In 69% of women of less than 30 years old, the tumor is detected in early stages. In these cases, cholecystectomy is the curative procedure, with a 90% survival at five years. According to our experience, cholecystectomies should be performed between 40 and 50 years of age in men and between 30 and 40 years in women. The prognostic factors that should be considered are symptoms associated to lithiasis, age, parity, obesity, size of stones and the size of the gallbladder. If the tumor is detected in early stages, the survival is good. The natural history of the disease would change significantly if all women with symptomatic stones were operated. PMID- 12587515 TI - [On the control of professional ethics]. AB - Professional colleges lost the ethical control of their members in 1980, when a free market economical model was implemented in Chile. Courts of justice have failed to accomplish this task. To recover such control, the Chilean Medical Association would need to modify its internal organization, separating its functions as a union from those required to control professional ethics. The health reform that is being set forth in the country, proposes a "new instance" for the control of professional ethics, within the Health Services and forming part of the Health Care Authority. PMID- 12587516 TI - [Use of case-parents trio for epidemiological studies of association between genetic polymorphisms and complex diseases]. AB - In case-control studies, spurious associations between allelic variants in candidate genes and disease may arise as a result of population stratification by ethnicity. In epidemiological terms, the ethnic group would act as a confounding factor if both the genotype frequency and the incidence/prevalence of a given chronic disease vary across ethnic groups. Population stratification is a key element in the study of population dynamics and the knowledge of the disparity in health-related conditions among strata has important social and public health implications. However, it would not be scientifically correct to conclude a causal gene-disease association in situations in which the association arises as a consequence of a particular population structure. Although the relevance and magnitude of the bias derived from this confounding, effect would be different depending on the population considered, there has been an increased use of study designs such as the case-parent study, that avoids the effect of population stratification by ethnicity. In this article, we review the analysis of case parent trios using unrelated affected cases as an epidemiological study design for testing association between genetic polymorphisms and disease. PMID- 12587517 TI - [Acute confusional syndrome associated to chlormezanone]. PMID- 12587518 TI - Correlation between hand preference and distance of focusing points of two eyes in the horizontal plane. AB - Relationships among hand preference, ocular dominance, and the degree of ocular shifting were studied in 78 right-handed and 16 left-handed subjects. Ocular dominance was assessed with the Miles test. The shifting degree of eye was assessed using a modified Miles test. The shifting distance of the right-eye was marginally greater, although significant, in the left-handers as compared to the right-handers. The shifting distance of the left-eye was greater in the right handers than in the left-handers. The distance of focusing points of two eyes in the horizontal plane was greater in the right-handers than in the left-handers. In the total sample, there was a significant negative Pearson correlation between hand-preference and the shifting distance of the right-eye, and there was a positive correlation between hand-preference and the shifting distance of the left-eye. In addition, there was a significant positive correlation between hand preference and the distance of focusing points of two eyes. These results suggest that hand preference may be related to the degree of ocular asymmetry. PMID- 12587519 TI - Heart rate and fluency performance among high- and low-anxious men following autonomic stress. AB - Given the well-known associations between the right frontal lobe and negative effective states, as well as the involvement of the right posterior region in the modulation of autonomic nervous system arousal, anxiety is thought to be associated with relative right hemisphere activity (or decreased left hemisphere activity). However, recent evidence suggests that the left hemisphere predominantly modulates parasympathetic efferents, while the right hemisphere predominantly modulates sympathetic efferents of the autonomic nervous system. Thus, the left hemisphere should also play a role in anxiety, as anxiety is associated with decreased parasympathetic activity (increased heart rate). In order to investigate this hypothesis, the present experiment examined the influence of a well-known autonomic stressor (cold pressor) on heart rate (HR), and verbal and nonverbal fluency performance. Fifteen high level (HI) anxious and 15 low level (LO) anxious men completed verbal and nonverbal fluency measures after they were asked to place their left hand in ice water for 45 s. HI anxious subjects demonstrated opposing HR patterns during performance of the verbal (increased HR) and nonverbal (decreased HR) fluency tasks following cold pressor stimulation. In addition, HI anxious men generated fewer words per minute on the verbal fluency task than did LO anxious men. Although the data are supportive of functional right hemisphere activity among anxious individuals, the contribution of decreased left hemisphere activity (i.e., reduced parasympathetic activity and diminished verbal fluency performance) is also discussed. PMID- 12587520 TI - Reduction of central poststroke pain with the selective serotonin reuptake inhibitor fluvoxamine. AB - To investigate the effect of the selective serotonin reuptake inhibitor (SSRI) fluvoxamine on central poststroke pain (CPSP), fluvoxamine (25 to 125 mg daily) was given to 31 patients. Although 3 patients dropped out within 1 week, 28 patients who received fluvoxamine for 2 to 4 weeks showed a significant reduction in the visual analog scale (VAS) for pain from 7.7 +/- 2.2 to 6.0 +/- 3.4 (p < .01). This improvement in VAS was significant in patients within less than 1 year after stroke, but not in those with a duration of more than 1 year. Zung's Self rating Depression Scale (SDS) was also significantly improved after treatment, but there was no significant correlation between the changes in VAS and SDS. Although this is not a double-blind, placebo-controlled trial, these results suggest that fluvoxarnine is useful for the control of CPSP regardless of depression when used relatively early after stroke. PMID- 12587521 TI - Does long-term use of valproate cause weight gain in prepubertal epileptic children? AB - In this experiment, we studied the effect of valproate (VPA) on weight gain, and serum leptin levels in prepubertal epileptic children receiving VPA. Our purpose was to determine whether or not long-term use of VPA causes weight gain in childhood, and to evaluate serum leptin levels in a group of prepubertal children receiving VPA. Our study included 15 patients (9 males, 6 females) with new diagnosed epilepsy and 16 healthy age-matched controls (9 males, 7 females). The subjects' ages ranged from 9 months to 12 years. Weight gain was noted in 9 (60%) of 15 patients in the study group, and 8 (50%) of 16 subjects in the control group (p > .05). There was no difference between the groups for body mass index (BMI) and serum leptin levels. Although higher serum leptin levels were found in the patients treated with VPA weight gaining (5.65 +/- 3.06 ng/ml vs. 3.28 +/- 1.69 ng/ml), we did not find a difference between the patients weight gaining and nonweight gaining (p > .05). While a significant correlation between BMI and serum leptin levels was found in the study group (r = .704; p = .003), it was not significant in the control group (r = .330; p = .211). In conclusion, our findings showed that long-term use of VPA did not cause weight gain in a group of prepubertal children receiving VPA and, parallel to this, serum leptin levels were similar in both the control and study group. PMID- 12587522 TI - Effects of exercise-induced fatigue with and without hydration on static postural control in adult human subjects. AB - Postural instability can be the result of various factors, including fatigue. Although it is well known that exercise-induced fatigue may be responsible for a decrease in performance, its effects on postural control, as well as those of hydration, have been relatively little explored. This study evaluated the effects of fatigue, with and without rehydration, on postural control in 10 healthy subjects who regularly practice sports activities. All subjects were submitted to three types of ergocycle exercises: maximal oxygen uptake (Vo2max) and submaximal exercises in no-hydrated and hydrated conditions at a power corresponding to approximately 60% of the Vo2max of each subject. Static posturographic tests were performed immediately before (control) and after exercises. The postural control performance decreased from the best to the worst: control, hydration, dehydration, and Vo2max. Fast Fourier transformation of the center of foot pressure showed three patterns of amplitude spectral density, with an increase of spectral amplitude for dehydration, more important for Vo2max conditions. Spectral amplitudes for control and hydration conditions were relatively similar. This hierarchy suggests that fatigue mainly alters muscular effectors and sensory inputs, such as proprioception, resulting in poor postural regulation. Moreover, fluid ingestion could be responsible for the preservation of muscular functions and of sensory afferences accurately regulating postural control. PMID- 12587523 TI - XY99-5038 promotes long-term survival of cultured retinal neurons. AB - We have previously reported that XY99-5038, a preparation from a specific formula of Traditional Chinese Medicine, could effectively inhibit hydrogen peroxide induced retinal cell death. In the present study, we investigated the possibility of XY99-5038 to prolong neuronal survival in a long-term retinal neuronal culture. Basic fibroblast survival factor (bFGF), a potent neurotrophic factor, was employed as comparable agent. Retinas of 0-2 days old Sprague-Dawley rats were isolated and dissociated. The cells were maintained in tissue culture for up to 9 weeks in a synthetic serum-free media. XY99-5038 (100 ng/ml) or a vehicle was added to culture every 3-4 days, starting at the first week of culturing. The number of cells were counted and compared for each time point and treatment. Cell viability was also determined by MTT assay, whereas apoptotic cell death was evaluated by the TUNEL assay. XY99-5038 treatment significantly reduced cell loss, increased cell viability, and inhibited apoptosis in this long-term retinal neuronal culture. Our data also show that the protective effect of XY99-5038 is more potent than that of bFGF. Our data suggest that XY99-5038 could be beneficial to the prolongation of neuron survival. PMID- 12587524 TI - Endogenous strychnine: description of hypo- and hyperstrychninergic state in relation to neuropsychiatric diseases. AB - Previous work from our laboratory has demonstrated the presence of endogenous strychnine in the mammalian brain and human serum samples. The present study examines the role of strychnine in neuropsychiatric disorders. Strychnine is synthesized from tryptophan. The blood levels of tyrosine, tryptophan, and strychnine were studied as also RBC membrane Na(+)-K+ ATPase activity. It was found that serum tyrosine levels were reduced and that tryptophan levels were elevated in all neuropsychiatric disorders studied with a reduction in RBC Na(+) K+ ATPase activity. Strychnine was present in significant amounts in the serum of patients with epilepsy, Parkinson's disease, and manic depressive psychosis. The presence of strychnine in significant amounts could be related to elevated tryptophan levels, suggesting the synthesis of these alkaloids from tryptophan. Na(+)-K+ ATPase inhibition present in most of the disorders could be related to increased depolarizing strychninergic transmission. The role of strychnine in the pathogenesis of these disorders, in the setting of membrane Na(+)-K+ ATPase inhibition, is discussed. PMID- 12587525 TI - Coordination in the dynamics of the brain as inferred from EEG analysis. AB - The present study observed the coordination between cortical areas during no task conditions as well as for the pathological condition of epilepsy, by application of the phase synchronization technique to the EEG signal in a multichannel recording. The index obtained from the phase entrainment investigation was properly scaled by a novel method to take into account the effect of nearest neighbor interactions. This scaled index was analyzed temporally to learn about the behavior of regional interactions in time. The results obtained not only corroborate earlier known results, but also give deeper insight into actual brain functioning. PMID- 12587526 TI - Nonlinear interdependence in neural systems: motivation, theory, and relevance. AB - In this article, we motivate models of medium to large-scale neural activity that place an emphasis on the modular nature of neocortical organization and discuss the occurrence of nonlinear interdependence in such models. On the basis of their functional, anatomical, and physiological properties, it is argued that cortical columns may be treated as the basic dynamical modules of cortical systems. Coupling between these columns is introduced to represent sparse long-range cortical connectivity. Thus, neocortical activity can be modeled as an array of weakly coupled dynamical subsystems. The behavior of such systems is represented by dynamical attractors, which may be fixed point, limit cycle, or chaotic in nature. If all the subsystems are perfectly identical, then the state of identical chaotic synchronization is a possible attractor for the array. Following the introduction of parameter variation across the array, such a state is not possible, although two other important nonlinear interdependences- generalized and phase synchronized--are possible. We suggest that an understanding of nonlinear interdependence may assist advances in models of neural activity and neuroscience time series analysis. PMID- 12587527 TI - The health-promoting behaviors and attitude towards menopause and hormone replacement therapy among women on dialysis. AB - The aim of this study was to assess the health-related behaviors, knowledge and attitude towards menopause and hormone replacement therapy (HRT) among women receiving dialysis treatment. The study population consisted of 44 women aged 55 75 years attending the dialysis unit. All women completed a 20-item questionnaire, which covered information on personal health-care habits, life style and attitude regarding menopause and HRT. The majority of participants were of desirable weight (body mass index < 25 kg/m2) and had a positive body image. About 80% had never smoked, and did not engage in regular physical activity. Over half underwent annual Pap smear, mammography, breast examination by a physician, and only a small minority performed breast self-examination. Surprisingly, 45% had used HRT in the past, but only 5% were current users. Although we expected postmenopausal women with a life-threatening disease to be more aware of personal health care than the general population, only a relatively small percentage of the study population safeguarded their health care, and used HRT. Stronger educational efforts may be needed to promote health-related behaviors and a positive attitude to health care, aging and menopause in this subgroup of patients. PMID- 12587528 TI - Effects of pentoxifylline in the prevention of ovarian hyperstimulation syndrome in a rabbit model. AB - Tumor necrosis factor alpha (TNF-alpha) and other cytokines have been implicated in the pathogenesis of ovarian hyperstimulation syndrome (OHSS). Pentoxifylline, a methylxanthine derivative, was found to inhibit TNF-alpha synthesis. The aim of this study was to evaluate whether the use of pentoxifylline would prevent the occurrence of OHSS in a rabbit model. Thirteen rabbits were divided into two groups. The first group (n = 6) were given pentoxifylline 15 mg/kg intravenously and the second group (n = 7) were given physiological serum 15 mg/kg before ovulation induction. Ovarian hyperstimulation was induced in rabbits by 200 IU equine chorionic gonadotropin on day 1 and 100 IU human chorionic gonadotropin on day 3. Blood samples were analyzed for TNF-alpha on days 1, 3 and 5. All animals were autopsied on day 6 to evaluate the ovarian weight, ascites formation and histopathological changes. There was no difference between groups regarding weight gain, ascites formation and plasma TNF-alpha levels (p < 0.05). Ovarian weight and number of ovulations were significantly lower in the pentoxifylline group than the control group (p < 0.05). Pentoxifylline did not prevent ascites formation despite the observed decrease in ovarian weight and number of ovulations in OHSS in a rabbit model. PMID- 12587529 TI - Insulin: does it induce follicular arrest in the rat ovary? AB - The goal of this study was to investigate histological changes of the rat ovary treated with either insulin or insulin plus human chorionic gonadotropin (hCG). The study was conducted in Celal Bayar University, School of Medicine, Animal Research Laboratory. Eighteen adult female Wistar rats were divided into three groups to receive saline, or insulin, or insulin plus hCG for 4 weeks. At the end of treatment the rats were sacrificed and the ovaries were evaluated with hematoxylin and eosin. There was no abnormal change in rats treated with saline. A thickened capsule, stromal hypertrophy and stromal cell hyperplasia, and no developing follicles, were observed in the insulin-only group. A thin capsule, developing follicles and corpora lutea, and normal theca cells and stroma were observed in the insulin-plus-hCG group. We conclude that insulin may lead to histological changes similar to stromal hyperthecosis and polycystic ovary syndrome, and may be one of the factors causing follicular arrest. PMID- 12587530 TI - Pre-eclampsia with fetal growth restriction: placental and serum activin A and inhibin A levels. AB - Activin A (beta A beta A) and inhibin A (alpha beta A) are dimeric glycoproteins secreted from early to term pregnancy in the maternal circulation. They circulate in higher amounts in women with gestational hypertension and/or pre-eclampsia, the most important gestational diseases also causing fetal growth restriction (FGR). Since no data are available in patients with pre-eclampsia and superimposed FGR, by using two-site immunoassays we evaluated serum activin A and inhibin A levels in serum samples collected from: healthy normotensive pregnant controls (n = 42); and women with pre-eclampsia with (n = 19) or without superimposed FGR (n = 21). In addition, by quantitative reverse transcriptase polymerase chain reaction the changes of alpha- and beta A-subunit mRNA expression in placentas collected from healthy controls (n = 7) and pre-eclamptic pregnancies with (n = 6) or without (n = 6) superimposed FGR was also investigated. Activin A and inhibin A serum levels were significantly higher in pre-eclampsia, and the presence of FGR did not significantly modify these concentrations. Similarly, inhibin-subunit mRNA levels in placentas from pre eclampsia were significantly higher than in controls, and FGR did not significantly affect this expression. The present data suggest that the increased placental expression of inhibin subunit mRNAs is part of the mechanism leading to increased serum activin A and inhibin A levels. PMID- 12587531 TI - Dose-dependent inhibition of mitochondrial ATP synthase by 17 beta-estradiol. AB - Mitochondria produce energy through oxidative phosphorylation. A key enzyme in this pathway is F0F1-ATP synthase, catalyzing ATP production from ADP and inorganic phosphate. Recently a subunit of F0F1-ATP synthase, oligomycin sensitivity-conferring protein, was identified as a new estradiol-binding protein. Estradiol could directly modulate mitochondrial ATP synthase activity through this subunit. In addition, intracellular ATP levels play a role in apoptotic death, which is an energy-dependent process requiring functioning mitochondria. Here we examined the effect of 17 beta-estradiol on F0F1-ATP synthase directly (in permeabilized cells) and in intact osteoclastic FLG 29.1 cells, a model of inducible apoptosis. The baseline F0F1-ATP synthase activity of FLG 29.1 cells was 4.485 nmol/min per mg. Estradiol rapidly inhibited F0F1-ATP synthase activity in the physiological range (half-inhibition concentration, IC50, of 30 nmol/l). With 1 nmol/l of estradiol, the inhibition was already significant (8-10% inhibition, p < 0.01) and with 100 nmol/l residual enzyme activity was only 15% (85% inhibition, p < 0.01). In addition, the effect of estradiol appeared to be directed towards F0F1-ATP synthase, since succinate sustained respiration, uncoupled from the electron transport chain, was unaffected by estradiol. We assayed F0F1-ATP synthase activity in FLG 29.1 cells during inducible apoptosis. No significant difference of ATP synthesis was detected in apoptotic cells versus controls. In conclusion, we showed a new non genomic effect of estradiol on a key mitochondrial enzyme, which thereby directly modulates cellular energy metabolism. PMID- 12587533 TI - Polish consensus: procedure in case of estrogen deficiency in women after breast cancer therapy. AB - Women surviving breast cancer in the postmenopausal period suffer from hormonal alternations with adverse effect on mental status and functioning of a number of organs and systems. Two thirds of these women had menopause before the diagnosis of breast cancer. In the remaining one-third ovarian failure is natural or induced by chemotherapy. Doctors cautiously approach the use of estrogen therapy in this group of patients. Their fears are not unsupported bearing in mind known epidemiological data exist linking breast cancer with the use of hormonal therapy. The purpose of this review is to evaluate current data on hormonal use and breast cancer risk. PMID- 12587532 TI - Effects of tamoxifen and estradiol on estrogen binding sites in the urogenital tract: an experimental study in the rabbit. AB - The aim of the study was to investigate the effects of estradiol and tamoxifen alone and in combination on the estrogen binding site status of the urogenital tract in the rabbit. Bilaterally ovariectomized rabbits were divided into four groups of six. Whereas the control group received no treatment, the remaining rabbits were treated with estrogen or/and tamoxifen. Cytosolic and nuclear fractions were isolated from the uterus, vagina, urethra and urinary bladder and used for binding site assay, by radioligand binding. The total weight of the rabbit vagina and uterus was increased significantly by both estradiol, tamoxifen and the combination of the two. The total weight of the urethra was increased only in the combination group. The cytosol binding site was downregulated by estradiol, tamoxifen and combination in the uterus, and in the vagina. Cytosol binding site in the urethra was not detected. The combination of estrogen tamoxifen markedly reduced the nuclear binding site in the urethra and decreased affinity of the nuclear binding sites in all three tissues. The data suggest that tamoxifen has a specific ability to modulate the transcriptional activity of the estrogen binding sites in the rabbit urogenital tract. PMID- 12587534 TI - Gene expression of adhesion molecules and matrix metalloproteinases in endometriosis. AB - Various types of cell adhesion molecules and matrix metalloproteinases (MMPs) seem to play an important role in the invasion process of endometriosis; however, limited investigation has focused on their gene expression in human peritoneal endometriotic lesions. A total of 63 endometriotic tissues were surgically obtained from 35 women with endometriosis, which included 43 pigmented and 20 non pigmented lesions. Gene expression levels of E-cadherin, alpha- and beta-catenin, MMP-2, MMP-9 and membrane-type 1 (MT1)-MMP in these endometriotic lesions were compared with those in normal eutopic endometrium obtained from 12 women without endometriosis. MMP-2, MMP-9 and MT1-MMP mRNA expression in pigmented lesions was significantly higher than that in normal endometrium (p < 0.05), whereas E cadherin, alpha- and beta-catenin mRNA expression was not suppressed in endometriotic lesions. There was a close correlation between MMP-2 or MT1-MMP and E-cadherin, alpha- or beta-catenin gene expression in 63 endometriotic tissues examined (p < 0.01). Immunohistochemical expression of E-cadherin, alpha- and beta-catenin in glandular epithelial cells was positive not only for all of seven cases with normal eutopic endometrium but also for 9 of 11 with ovarian endometriosis. MMP expression in ectopic endometrium was much greater than that in eutopic endometrium. These results suggest that endometriotic tissues expressing MMPs might be invasive and simultaneously possess cell-to-cell adhesion property in pelvic peritoneal foci. PMID- 12587535 TI - Ovarian hyperstimulation-like syndrome after administration of triptorelin to a woman with endometriosis. AB - Ovarian hyperstimulation after a single dose of gonadotropin-releasing hormone (GnRH) analog is a rare phenomenon. A case of ovarian hyperstimulation-like syndrome after sole administration of triptorelin (Decapeptyl 3.75 mg) is reported in a woman who had undergone surgery for an endometriotic cyst. After administration of the drug, abdominal pressure increased with nausea and diffuse pelvic pain. Ultrasound examination showed bilateral enlargement of the ovaries (right 74 x 62 mm, left 62 x 53 mm), more than 10 follicles ranging in diameter from 15-25 mm, proliferative endometrium 7 mm thick and fluid in the Douglas pouch up to 25 x 23 mm thick. Estradiol plasma level was in the normal range. The syndrome spontaneously resolved in the course of treatment and a spontaneous pregnancy occurred when the triptorelin effect disappeared. PMID- 12587537 TI - Pregnancy-associated Cushing's syndrome secondary to a luteinizing hormone/human chorionic gonadotropin receptor-positive adrenal carcinoma. AB - Cushing's syndrome occurring during pregnancy is frequently due to an adrenal neoplasm. Adrenal gland tumors occasionally respond to luteinizing hormone (LH) or human chorionic gonadotropin (hCG). We report a case of Cushing's syndrome during and following pregnancy due to an adrenal carcinoma which expressed the LH/hCG receptor. The presence of these receptors may have led to the growth and function of the tumor during pregnancy. PMID- 12587536 TI - Circulating nitric oxide changes throughout the menstrual cycle in healthy women and women affected by pathological hyperprolactinemia on dopamine agonist therapy. AB - The aim of the study was to evaluate circulating nitric oxide (NO) levels throughout ovulatory cycles in healthy women and women under long-term treatment with dopamine agonists. Fifty women (aged 32.5 +/- 1.2 years) affected by pathological hyperprolactinemia (prolactin (PRL)-secreting microadenoma, 63%; idiopathic, 19%; 'empty sella', 12%; and PRL-secreting macroadenoma, 6%) and on dopamine-agonist therapy (range 1-10 years) were studied; 37 healthy women (aged 30.4 +/- 1.4 years) served as a control group. Blood samples were collected on days 7, 14 and 21 of the menstrual cycle in order to assay NO, PRL, 17 beta estradiol and progesterone. In all subjects, ovulatory cycles were recorded. PRL levels were comparable between the two groups and significantly rose during the luteal phase. NO levels recorded throughout the menstrual cycles of healthy controls were significantly higher than those recorded in subjects treated with dopamine-agonist; NO levels in the latter were no different from those recorded in non-treated, non-ovulatory hyperprolactinemic women. However, in both healthy controls and dopamine-agonist-treated women, NO was negatively correlated with progesterone concentration and significantly reduced on day 21. In dopamine treated patients, NO levels did not correlate with the dose or the duration of dopamine-agonist therapy. We conclude that, in our hyperprolactinemic women on therapy, physiological NO secretion is not fully restored, despite restoration of ovulatory cycles by dopamine-agonist therapy. PMID- 12587538 TI - Hyponatremia and hypoglycemia in acute Sheehan's syndrome. AB - We report the case of a 23-year-old Saudi Arabian woman who presented to the medical intensive care unit with severe hyponatremia and hypoglycemia following a Cesarean section delivery complicated by hemorrhage due to disseminated intravascular coagulopathy. She was treated successfully for adrenal insufficiency acutely, and was later discharged on hormone replacement therapy. To our knowledge, this is the first case report of acute Sheehan's syndrome presenting with both hyponatremia and suggestive hypoglycemia. Pituitary necrosis is an uncommon complication of peripartum hemorrhagic shock. Since the initial description by Sheehan in 1937, the incidence of the syndrome has gradually declined through improved management of hemodynamic complications leading to the infarction of the gland. There are many studies describing complications of late Sheehan's syndrome; however, relatively few contain descriptions of the acute phase. In addition, the diagnosis of this syndrome is often determined after resolution of the acute process with resultant lack of data regarding immediate endocrine and imaging abnormalities. In this report, we describe the complete endocrine and imaging assessment of a patient presenting in critical condition due to necrosis of the pituitary gland in the immediate postpartum period. PMID- 12587539 TI - [New developments in the diagnosis and therapy of acute pulmonary embolism]. AB - Despite the widespread use of thromboprophylaxis, pulmonary embolism (PE) remains one of the most common life-threatening disorder. In fact, the prevalence of detected postmortem PE has not diminished during the last 3 decades. The review summarizes the most important data on the prevalence of acute pulmonary embolism and the difficulties of diagnosis and differential diagnosis. Anticoagulant-, thrombolytic- and surgical therapy of pulmonary embolism is detailed. PMID- 12587540 TI - [Immunohistologic changes of the intestinal mucosa in chronic inflammatory bowel diseases]. AB - In the pathogenesis of inflammatory bowel disease abnormally increased immune reactions in the intestinal mucosa play a basic role. The revealing of these reactions was facilitated by the rapidly spreading immunohistological methods in last decades. This review discusses in detail the characteristic distribution of lymphocyte subsets and plasma cells in Crohn's disease and ulcerative colitis. Results indicating increased expression of adhesion molecules are also presented, as well as the different patterns of cytokine production in Crohn's disease and ulcerative colitis. The rapid epithelial turnover developing as a consequence of increased rate of proliferation and apoptosis in inflammatory bowel disease is also shown. The epithelial expression of MHC II antigen in colonic mucosa which is not observed in healthy subjects is also discussed. In addition, the pathogenetic role of lost oral tolerance in the development of inflammatory bowel is reviewed in detail. PMID- 12587541 TI - [Incidence of diabetes mellitus in patients with malignant tumors of the oral cavity]. AB - Diabetes mellitus affects 5% of the population of Hungary. Its complications in the oral cavity include parodontosis and changes in the oral mucosa (leukoplakia, lichen oris and glossitis). The incidence of leukoplakia (the most frequent precancerous state in the oral cavity) is 2.2% in the normal population, 6.2% among diabetics, and 11.5% among diabetic who smoke. The various forms of leukoplakia in the oral cavity become malignant in 5-20% of the cases, while the erosive, ulcerative lesions do so in 15-20% of the cases. These data led us to examine the blood glucose levels in 318 patients with histologically confirmed malignant tumours in the oral cavity. Diabetes mellitus was found in 56 of them (17.5%), the blood glucose level being elevated in 31 cases (9.75%). The fact that the incidence of diabetes mellitus among these patients was more than 3 times higher than that for the normal population demonstrates that further studies should be performed on the connection between diabetes mellitus and malignant tumours in the oral cavity. PMID- 12587542 TI - [Evaluation of results in transcervical hysteroscopic myoma resection]. AB - INTRODUCTION: The benign uterine fibroid is one of the most common cause for complains of females. The most important aim of the medicine nowadays is to treat the patient with minimally invasive methods, saving the organ. OBJECTS: Authors evaluate the efficacy of the transcervical hysteroscopic fibroid resection (TCRM) among their patients. MATERIALS AND METHODS: Authors performed 66 transcervical hysteroscopic fibroid resection (TCRM) in 61 patient in a six-year period. Based on clinical computed database and mailed questionnaires 51 patients were followed up. RESULTS: 30 out of 51 patients showed no symptoms after the procedure, in 14 cases subsequent fibroid was detected, while in 7 cases other type of complains occurred not related to the myoma. During the procedures complications appeared only in 4 cases, one perforation and three bleeding episodes occurred. CONCLUSIONS: Based on the data authors emphasize that TCRM is a safe and excellent method in treating submucosal fibroids. PMID- 12587543 TI - [Use of the Beck depression scale to screen patients treated at day hospitals for mobility disorders]. AB - OBJECTIVE: Authors analysed the prevalence of depressive symptomatology among the patients of a day time rehabilitation department for mobility disorders. METHODS: Beck's Depression Inventory was used as a screening instrument. 57 subjects completed the inventory both at admission and discharge. RESULTS: 33 subjects of 57 showed depressive symptoms at admission and 13 persons at discharge. The number of patients with mild depressive symptomatology decreased from 24 to 10, with medium level depression from 5 to 2 and with severe depression from 4 to 1. None of them have taken antidepressive drugs. CONCLUSIONS: These results confirm the high importance of the proper psychological conduction of patients with depressive symptoms. PMID- 12587544 TI - [Proportion of alcohols of cholesterol, glycerol and double bonds of fatty acids in blood serum and lipoproteins. Diagnostic significance of hypercholesterolemia]. AB - A new method in quantitative determination of double bonds (DB) in the pool of fatty acids (FA) has been worked out by ozone titration after extraction of lipids according to Folch. A correlation was observed among DB content, the level of cholesterol alcohol (CS, r = +0.612; p < 0.001) and the level of glycerol (GL) alcohol (r = +0.392; p < 0.01). The grown-ups with ischemic heart disease and children have a constant ratio (5.05 +/- 0.18) DB:CS ratio. PMID- 12587545 TI - [A new method for diagnosis of liver damage]. PMID- 12587546 TI - [Optimization of determination of phenylalanine concentration in blood samples of newborns]. PMID- 12587547 TI - [A simple method for estimation of concentration of vitamins E and A in low density lipoproteins]. AB - The aim of the study is to work out a new easy method for determination of fat soluble E and A vitamins concentration in low density lipoproteins (LDL). The method was worked out in the blood serum of 60 healthy men aged 35-55 with monolipidaemia and 55 men aged 45-65 with dislipoproteinemia (DLP). The new method consists of LDL isolation from serum by heparin precipitation method and determination of alpha-tocopherol and retinol concentrations by spectro fluorimetrical method. The new method is informative for estimation of LDL antioxidant system, easy and fast to perform. The new method features a high reproduction and strong positive correlation between levels of E and A vitamins in the blood serum by spectro-fluorimetrical method and levels of E and A vitamins in LDL by our method. The use of both methods shows reliable reduction in E and A vitamins concentration in the blood serum and in LDL in men with DLP type II compared with E and A vitamins concentration in men with normolipidaemia. PMID- 12587548 TI - [End point and kinetics. Pluses and minuses]. AB - There are results of laboratory quality control in determination of transminase activity by two methods--by end point and by kinetic method. Based on the performed dispersion analysis, a conclusion was made that despite the correspondence of both methods to the order # 45 of RF Ministry of Health, the determination of ALT and AST activity by end point does not register the moderately pronounced increase in the level of these enzymes. This point makes it impossible to use the data obtained in the analysis of transminase activity by end point to verify and monitor syndrome of endogenous intoxication. PMID- 12587549 TI - [Comparison of determination methods and application of a computer program using a screening coordinate system to interpret hemoglobin A1c in patients with diabetes types 1 and 2]. AB - The aim of the work is to compare methods of hemoglobin A1c determination in ratio with basic metabolism indices using computer program with graphic interpretation of results in patients with diabetes of 1 and 2 types. It was found out that more specific hemoglobin determination by isoelectrofocusing method more completely reflects the state of carbohydrate metabolism in diabetic patients. The importance of hemoglobin A1c determination in case of 2 type diabetes is particularly emphasized. PMID- 12587550 TI - [Differences in information on biochemical indices among different population groups under conditions of a comprehensive polyclinic examination]. PMID- 12587551 TI - [Morphofunctional condition of peripheral blood erythrocytes in late vascular complications of diabetes type 2 (literature review)]. PMID- 12587552 TI - [Cytological examination of liver damage (lecture)]. PMID- 12587553 TI - [Proliferative activity of lymphocytes of peripheral blood in infectious mononucleosis in children]. AB - Changes in proliferative activity of lymphocytes from peripheral blood may serve an additional diagnostic criterion of etiological variants of infectious mononucleosis (IM)(viral and bacterial). In MN of viral etiology in evident disease the ability of lymphoid cells to blasttransformation in the presence of PHA is low vs healthy children. In convalescence the changes persist, proliferative activity of cells increase in stimulation by lipopolysaccharide. Later blast-transformation of lymphocytes return to normal. In bacterial IM proliferative activity of PHA- and LPS-stimulated lymphocytes is normal. PMID- 12587554 TI - [Quality problems in quantitative determination of hemoglobin in blood]. AB - Sources of errors in determination of hemoglobin were found out. The results were demonstrated to have comparison hemoglobin determination by hemiglobincyanide and hemichrome methods; this or that set of agents is of no significant advantage; to properly determine hemoglobin in the blood it is important to have calibration specimens and high quality control hemoglobin solutions; the equipment in use should be checked and calibrated; calibration hemiglobincyanide specimens produced at SPA "Renam". "Bak. preparations" and hemichrome specimens produced at JSC "Vector-Best" provide high precision of hemoglobin determination; the results of hemoglobin determination in control solutions obtained in autoanalyzers and manually, are comparable. PMID- 12587555 TI - [Integral indices of peripheral blood leukogram in the estimation of non-specific immunological reactivity in patients with ischemic heart disease]. AB - With the help of differential blood count analysis and velocity of erythrocyte sedimentation (VES), 30 healthy persons (donors), 30 patients with chronic ischemic hearty disease (IHD) and 34 patients with acute myocardial infarction (AMI) underwent the procedure of calculation for leukocyte index (LI), leukocyte intoxication index (LII), leukocyte shift index (LSI), leukocyte and VES ratio (LVESR), leukocytic and granulocytic index (LGI), general index (GI), neurophil lymphocyte ratio (NLR), neurophil-monocyte ratio (NMR), lymphocyte-monocyte ratio (LMR) and lymphocyte-eosinophil ratio (LER). Unlike healthy people, patients with chronic IHD had higher indices of LVESR, GI and LER while patients with AMI had increasing indices of LII, LSI, NLR, LER and decreasing indices of LI, LGI, GI, LMI. In case of AMI compared with chronic IHD, average indices of LII, LSI, NLR were higher and indices of LI, LGI, LVESR, GI, LMR were lower. The obtained results show expansion of possibilities to get information about the state of non specific immunologic reactivity in patients with various IHD forms with the help of integral indices in blood leukogram. PMID- 12587556 TI - [Laboratory diagnosis of cytomegaloviral infection in patients with anemia and hemoblastosis in the Omsk region]. AB - Research was carried out in 335 blood specimens of patients in the age of 3-35 y.o. in order to optimize diagnosis and treatment of such patients with aplastic anemia and hemoblastosis who got hemotransfunction to eliminate cytomegaloviral infection (CMVI). IgM were found out in 37.9% cases (2.8 times higher than in donors), low-avide IgG--in 44.8%. "early" proteins CMV--29.9% and DNA--in 36.8% cases. Concerning the DNA presence, preference was given to research of leucocytic suspension compared with blood serum. Of 28 children of 3-13 y.o. with anemia being seropositive in CMV, IgG antibodies were detected in 13 children while IgM antibodies to Parvovirus B19 were found in 10 children. 7 children with a grave form of disease showed combined infection of Parvovirus B19 and CMV with activation signs. It is not excluded that parallel influence of Parvovirus B19 on erythrocytic hemopoiesis growth and that of CMV on lymphocytic-monocytis cells aggravates immunodeficiency and promotes development of infection complications. PMID- 12587557 TI - [An experience of quality evaluation of Corynebacterium species]. PMID- 12587558 TI - [State of the art and outlook for the laboratory diagnosis of brucellosis]. PMID- 12587559 TI - [Obtaining diagnostic fluorescent monoclonal immunoglobulins against the V. cholerae 0139-serovar]. AB - Below is given a procedure of the obtaining diagnostic fluorescent monoclonal immunoglobulin to detect cholera vibrios of O139 serovar. While obtaining preparations it was managed to determine optimal FTTS-MKA ratio, duration of their conjugation, series of fluorochrome. Test specimens of fluorescent monoclonal immunoglobulin provides intensive glow of V cholerae O139 cells in the working dilution 1:16-1:32. Tests of diagnostic FTTS-MKA on the great number of homologic and heterologic strains showed their strict specificity and high sensibility as to cholera vibrios of O139 serogroup. PMID- 12587560 TI - Skin cancer prevention and control. PMID- 12587565 TI - Serious neurological conditions following pertussis immunization: an analysis of endotoxin levels, the vaccine adverse events reporting system (VAERS) database and literature review. AB - The purpose of this study was to determine the potential risks for the development and outcome of serious neurological illnesses following whole-cell DTP vaccination and also to determine if the switch to using acellular DTaP vaccine in the US has had any effect on the incidence rate of serious neurological illnesses following vaccination. This study used the Limulus amebocyte lysate (LAL) endotoxin assay to determine the levels of endotoxin in various commercially available whole-cell and acellular DTaP vaccines, analysed the Vaccine Adverse Events Reporting System (VAERS) database to determine the clinical effects of the use of whole-cell DTP and acellular DTaP vaccines in the US and reviewed recently published pertinent studies that analysed the incidence rates of serious neurological illness following whole-cell DTP and acellular DTaP vaccines. The results indicated that whole-cell DTP vaccine contained high levels of endotoxin and was statistically significantly more reactogenic than acellular DTaP vaccine. The presence of bias in the VAERS database was not borne-out. The recommendation by the American Academy of Paediatrics to use acellular DTaP vaccine for the entire childhood vaccination schedule beginning in 1996 and the absence of the availability of whole-cell DTP in the US beginning in 2001 seems well justified based upon the results of this study. PMID- 12587566 TI - Quranic description of gametes. AB - There are several important characteristic properties of reproductive fluid. Gametes are the reproductive cells consisting of sperms in males and ovum in females. These cells combine to form Zygote in the process of fertilization. The characters of Gametes have been narrated by Holy Quran in a very well-suited and selected terminology. This paper enumerates various Quranic terms related to gametes, and then discusses their scientific significance. PMID- 12587567 TI - A brief on Ayurvedic manuscripts preserved at Utkal University, Bhubaneswar. AB - Amongst different institutions, "Parija Library" of Utkal University, Bhubaneswar has about 101 palm-leaf manuscripts (mss.) on Ayurveda to its credit. Most of these mss. have not been reported earlier. Herewith a brief description of these mss. has been made with the intention of bringing it out for the Ayurvedic fraternity and to enrich the literary stock of Ayurveda. PMID- 12587568 TI - Breast-conserving surgery for breast cancer. PMID- 12587569 TI - Religious practices in South India to cure diseases. AB - There is belief in South Indian villages that epidemics are due to evil spirits surrounding the villages. The propitiating of Gods and Goddesses to get rid of diseases in these villages are discussed. The Christian and Muslin traditions to cure the diseases by practising religion are also discussed. PMID- 12587570 TI - Breast-conserving surgery for breast cancer. PMID- 12587571 TI - [Strategies to reduce the development of bacterial resistance]. PMID- 12587572 TI - [Fungal infection in hematology]. PMID- 12587573 TI - Epidemiology and multiple sclerosis. PMID- 12587574 TI - Digoxin for the treatment of heart failure. PMID- 12587575 TI - Digoxin for the treatment of heart failure. PMID- 12587576 TI - Residents' work hours. PMID- 12587577 TI - Residents' work hours. PMID- 12587578 TI - Residents' work hours. PMID- 12587579 TI - Elecampane and Job's disease. AB - Elecampane (Inula Campana) sometimes referred to as Inula helenium is famous for its successful use as a medication by Prophet Job who had chronic sore boils. Hippocrates described it as a good and effective cure for chronic skin eruptions and itch etc. Elecampane is also reputed to relieve all kinds of pains, especially those arising from chills, and animal bites. In traditional medicine, Elecampane was and is still used in different forms for the cure of many diseases. PMID- 12587580 TI - Look before you quote. PMID- 12587581 TI - Is war necessary? PMID- 12587583 TI - Is war necessary? PMID- 12587582 TI - Is war necessary? PMID- 12587584 TI - Is war necessary? PMID- 12587585 TI - Pain in infants. PMID- 12587589 TI - On a first-name basis. PMID- 12587590 TI - What's happened to American health care? PMID- 12587591 TI - Patients benefit from nursing follow-up. PMID- 12587592 TI - Women constitute half of the cases of HIV and AIDS. PMID- 12587593 TI - New guideline for hand hygiene. Soap and water are still necessary. PMID- 12587594 TI - Diabetes management. PMID- 12587595 TI - Commentary: Matched cohorts can be useful. PMID- 12587596 TI - Effect of computerised evidence based guidelines. Challenge should not be abandoned. PMID- 12587598 TI - A valuable and timely analysis of research publications in 4 premiere physical therapy journals. PMID- 12587599 TI - Effect of computerised evidence based guidelines. Opportunity was missed. PMID- 12587601 TI - Posttransplant diffuse large B-cell lymphoma of "lymphomatoid granulomatosis" type. AB - The exact relationship between lymphomatoid granulomatosis (LyG) and posttransplant lymphoproliferative disorders (PTLDs) is not clear. Both are observed in immunodeficient patients and are Epstein-Barr virus driven. These disorders are, however, considered distinct based upon the immune response elicited; LyG is T-cell rich while PTLDs are T-cell poor. We describe a case of LyG-type diffuse large B-cell lymphoma (DLBCL) in a lung transplant recipient. The unusual features include rare occurrence of LyG in a posttransplant setting, systemic involvement by LyG variant of DLBCL in a solid organ transplant recipient, paucity of T-cells in this LyG type lymphoma, and subcutaneous panniculitic pattern in a B-cell lymphoproliferative disorder. This first report of systemic LyG variant of DLBCL in a posttransplant setting has features suggesting similarities and overlap between LyG and PTLD. PMID- 12587602 TI - Postmortem measurement of serum procalcitonin concentration in Waterhouse Friderichsen syndrome. PMID- 12587603 TI - A limited form of Churg-Strauss syndrome presenting as acute abdominal catastrophe. PMID- 12587604 TI - Evidence-based laboratory medicine: addressing bias, generalisability and applicability in studies on diagnostic accuracy: the STARD initiative. PMID- 12587605 TI - [Reflux-like dyspepsia in obese patients]. AB - The aim of this study was to assess, with a specially prepared questionnaire, the prevalence of reflux-like dyspepsia in a population of 40 patients morbidly obese, Body Mass Index (BMI) 46.2+/- 1.7 kg/m2, comparing the results with those deriving from the analysis of 20 healthy volunteers (BMI 20.9+/-1.7 kg/m2). The prevalence of esophageal symptoms in all obese patients was 45%, versus 15% in controls. There was a significant direct correlation between obesity and reflux like dyspepsia (p<0.05). Patients who are morbidly overweight should be encouraged to lose weight, as the very first step in fighting Gastro-Esophageal Reflux Disease. PMID- 12587606 TI - [Reduction of antibiotic resistance of Enterococcus faecalis in infections of the genitourinary tract]. AB - Antimicrobial susceptibility levels of Enterococcus faecalis isolated from urine were followed up at our University Hospital during three years (1999-2001), in order to evaluate the trend of antibiotic resistance of this common community acquired pathogen of the genito-urinary tract. One thousand and 249 consecutive strains were examined, and a surprisingly low resistance profile was detected. As a result, penicillin , ampicillin, nitrofurantoin and piperacillin may still represent first-line agents in our area when community-acquired urinary tract enterococcal infections is of concern, pending in vitro susceptibility studies. PMID- 12587607 TI - [Diagnosis of malaria with immunochromatographic test The Novara experience]. AB - Microscopic detection of parasites has been the reference standard for malaria diagnosis for decades. Recently rapid antigen assays provide an effective tool for the detection of malaria in symptomatic patients. In Novara Maggiore della Carita Hospital we evaluated the efficacy of the ParaSight F test in patients with malaria. PMID- 12587608 TI - Epidemiology of peripheral arterial disease in women. AB - BACKGROUND: Peripheral arterial disease is a common disease, which increases with age and presence of vascular risk factors. The extended longevity in industrialized nations coupled with the expanding elderly female population is predicted to lead to an increase in the prevalence of this condition. Little attention has been focussed on gender differences in peripheral arterial disease, or its epidemiology in women. METHODS: MEDLINE search of English-language reports published between 1966 and 2002 and search of references of relevant papers. RESULTS: Across various populations of women with different ages and risk factor levels, the prevalence of peripheral arterial disease ranged between 3% and 29%. Diagnosis in women using a sensitive and specific non-invasive test, the ankle brachial index, detects about 3-5 times the cases than those diagnosed by history of intermittent claudication alone. Contrary to earlier beliefs, prevalence of peripheral arterial disease is similar in women and men, and women may have more asymptomatic disease. Importantly, women with peripheral arterial disease have 2 4 fold increases in cardiovascular morbidity and mortality. Risk factors for peripheral arterial disease appear to be similar in men and women, however relative risks vary somewhat. CONCLUSIONS: Performing non-invasive testing (e.g. ankle-brachial index) can better diagnose peripheral arterial disease in women than history alone. These women share risk factors with other vascular diseases (coronary and cerebrovascular disease) and should undergo risk factor modification to reduce their cardiovascular morbidity and mortality. More research is needed including aggressive risk factor management in women with asymptomatic peripheral arterial disease. PMID- 12587609 TI - The association of fatty liver and diabetes risk. AB - We examined whether fatty liver, as diagnosed with abdominal ultrasonography, is an independent risk factor for diabetes mellitus during 10 years of follow-up. A total of 840 subjects (467 men and 373 women) were followed for the entire 10 years. The criteria for being non-diabetic were having no history of diabetes, having a fasting plasma glucose level of less than 110 mg/dl and a serum hemoglobin A1c level of 6.4% or less. We indicated that every examine received all examinations after 12 hours of fasting. Well-trained technicians performed abdominal ultrasonography. Although univariate analysis revealed that the presence of fatty liver was related to hyperglycemia 10 years later, multiple logistic regression analysis did not support this finding. In the multiple logistic regression analysis fasting plasma glucose levels at the baseline and age were significantly related to hyperglycemia (odds ratio [OR] = 1.16, 95% confidence interval [CI]: 1.11-1.21, OR = 1.07, 95% CI: 1.01-1.14, respectively). Fatty liver was not an independent risk factor for hyperglycemia in our follow-up study 10 years after the first diagnosis. The high fasting plasma glucose levels were a risk factor for diabetes, even in the normal range. PMID- 12587610 TI - Menstrual and reproductive factors and risk of lung cancer among Chinese women, Eastern Gansu Province, 1994-1998. AB - There are suggestions that women have higher relative risks of lung cancer than men given similar levels of tobacco exposure, implying that sex-related hormones may influence lung cancer risk. We evaluated the association between lung cancer risk and menstrual and reproductive factors on a subset of self-responding females in a population based case-control study in Eastern Gansu Province, China, 1994-1998. The analysis included 109 incident lung cancer cases and 435 controls selected from the census list and frequency matched to cases on age and prefecture. Odds ratios were lower for later ages at menarche (trend, P = 0.015) and later ages at menopause (trend, P = 0.074). Despite limitations, these findings suggest a possible role of hormones in the etiology of lung cancer in females. PMID- 12587611 TI - The relationship between smoking habits and serum levels of 8-OHdG, oxidized LDL antibodies, Mn-SOD and carotenoids in rural Japanese residents. AB - This study investigated the relationship between smoking habits and serum levels of 8-OHdG, oxidized LDL antibodies (oLAB), Mn-SOD, and carotenoids. Subjects were 79 males (mean age +/- standard deviation; 62.1 +/- 10.0 years) and 79 females (60.3 +/- 10.3 y) who attended a health examination screening in the town of Hokkaido, Japan. Serum 8-OHdG, Mn-SOD, and oLAB levels were measured by ELISA and serum carotenoids levels were measured by HPLC. Smoking habits were assessed by public health nurses using a questionnaire. Serum 8-OHdG levels were significantly higher in males than in females. On the other hand, serum levels of beta-carotene, alpha-carotene, beta-cryptoxanthin, and zeaxanthin and lutein were significantly lower in males than in females. Serum beta-carotene, beta cryptoxanthin, and zeaxanthin and lutein were significantly lower in males who were current smokers, compared to non-smokers. Serum 8-OHdG levels were also significantly higher in current smokers. Furthermore, in males, serum oLAB and beta-carotene levels were significantly and negatively correlated with the number of cigarettes smoked per day. Serum Mn-SOD levels were unrelated to smoking habits in males. In conclusion, this preliminary study suggests that cigarette smoking increases serum 8-OHdG levels and reduces serum levels of oLAB and carotenoids, such as beta-carotene, beta-cryptoxanthin, and zeaxanthin and lutein in healthy male subjects. Serum levels of 8-OHdG, oLAB, and carotenoids may be useful biomarkers of oxidative conditions affected by smoking. PMID- 12587612 TI - Gender and age differences in lifestyle factors related to hypertension in middle aged civil service employees. AB - The aim of this study is to identify lifestyle factors related to hypertension in man and woman workers, and to investigate age and gender differences in the relationships of the factors. From 6,000 civil service employees (4,937 men and 1,063 women) aged 40-69 years, information on lifestyle-related factors such as stress, exercise habits, preference for salty taste, alcohol drinking and smoking habits, and body mass index, as well as age and family history of hypertension was obtained through self-administered questionnaires in 1997. Hypertension was defined as either a systolic blood pressure > or = 140 mmHg, a diastolic blood pressure > or = 90 mmHg, or undergoing treatment for hypertension, and was present by 37.0% in men and 19.6% in women. Only body mass index was a significant lifestyle-related risk factor common to both genders with an odds ratio and its 95% confidence interval in parentheses of 2.2 (2.0-2.5) for men and 3.2 (2.3-4.6) for women. Men and women who preferred salty taste showed multivariate adjusted odds ratios of 0.9 (0.8-1.1) and 1.5 (1.1-2.2) for hypertension, respectively. In the stratified subanalysis, women aged 50 years and over had a significant odds ratio of 2.7 (1.5-4.9), whereas women aged 40-49 years and men of all age classes failed to show significant relationships. Salt intake was suggested to be a key factor for hypertension particularly for women after menopause. PMID- 12587613 TI - Associations of lifestyle factors with bone mineral density among male university students in Japan. AB - To investigate associations of lifestyle factors with bone mineral density among young men in Japan, we measured bone mineral density of the second metacarpal bone in 143 male university students, aged 18-22 years, by the computed X-ray densitometry. The subjects completed a lifestyle questionnaire including a quantitative food frequency questionnaire. Their mean+/-standard deviation of bone mineral density was 2.61+/-0.23 mmAl. Body mass index (Spearman's rho=0.232, p=0.006), daily walking time (rho=0.186, p=0.028), and milk consumption at junior (rho=0.250, p=0.003) and senior (rho=0.195, p=0.020) high school were significantly correlated with the bone mineral density. For nutritional variables, the bone mineral density was positively correlated with energy adjusted intakes of calcium (Pearson's r=0.302, p=0.0002), potassium (r=0.265, p=0.001), saturated fatty acids (r=0.211, p=0.011), and magnesium (r=0.173, p=0.039), and with those of milk and dairy products (r=0.228, p=0.006) and fruits (r=0.205, p=0.014), while being negatively associated with energy-adjusted noodle consumption (r=-0.185, p=0.027). The positive correlation of milk consumption at junior high school with the bone mineral density was not materially altered by adjustment for the body mass index, calcium intake, and walking time. Single-life students had lower bone mineral density compared with those lived with families (p=0.044). Bone mineral density could be increased by modifying dietary habits in young men. PMID- 12587614 TI - An ecological study of the relations between the recent high suicide rates and economic and demographic factors in Japan. AB - The male suicide mortality in Japan reached the highest in 1998, since statistics were first recorded in 1899. We investigated whether the recent high suicide rates were associated with economic and demographic factors, using the Pearson's correlation and multivariate regression analyses. The annual numbers of suicide deaths, economic variables, and the proportion of elderly people between 1995 and 2000 were obtained for all the 47 prefectures in Japan. The correlation analyses showed that the male SMR of suicide was mostly associated with the economic variables and the proportion of the elderly men. The female SMR of suicide was associated with the job application rate, and the proportion of elderly women in several years. The multivariate regression analyses showed that the male SMR of suicide was mostly associated positively with the job application rate and the proportion of elderly men and negatively with the amount of household savings and public assistance rate. The largest proportion of the variance of the male SMR of suicide was explained by the set of independent variables in 1998. The goodness of-fit of the model concerning the female SMR of suicide was poor. PMID- 12587615 TI - Cost performance and efficacy of off-pump coronary artery bypass grafting. AB - Off-pump coronary artery bypass grafting (Off-Pump CABG) may provide an alternative form of surgical revascularization by avoiding the unwanted complications of cardiopulmonary bypass, particularly in high-risk patients. To clarify the efficacy and cost performance of Off-Pump CABG, we studied the postoperative course of Off-Pump CABG and compared it to On-pump coronary artery bypass grafting (On-Pump CABG). From Aug. 1998 to Feb. 2002, twenty-eight patients who had preoperative complications such as cerebral vascular disease (11), chronic renal failure (4), atheromatous aorta (4), one lung (1), severely impaired left ventricular function (6), re-do CABG (1), and cancer (1) underwent Off-Pump CABG. Another thirty-six patients who underwent On-Pump CABG served as a control. The Off-Pump CABG patients were almost the same age as the On-Pump CABG patients (68 +/- 8 vs 64 +/- 8 years, ns). The Number of grafts was similar in both groups (2.6 +/- 1.0 vs 2.9 +/- 1.0, ns). Peak CK, peak CKMB, peak LDH, and peak GOT release were significantly lower in the Off-Pump CABG group compared with the On-Pump CABG group. Graft patency rates were similar in both groups (98% in Off-Pump CABG vs 98% in On-Pump CABG). The total cost for surgery and patient care was significantly lower (p < 0.0001) in the Off-Pump CABG group (dollar 21000 +/- 7000) compared with the On-Pump CABG group (dollar 33000 +/- 4200). Off Pump CABG is less invasive to the myocardium, is less expensive, and has a similar efficacy in comparison with On-Pump CABG. PMID- 12587616 TI - Use of the laryngeal mask airway in combination with regional anesthesia facilitates induction and emergence from general anesthesia in patients undergoing colorectal surgery. AB - The laryngeal mask airway (LMA) is selected as an alternative to the endotracheal tube (ETT) when rapid recovery from general anesthesia is considered. However, the clinical significance of this airway for abdominal surgery is unclear. Thus, we evaluated whether the LMA, in combination with regional anesthesia, facilitates the induction of and emergence from general anesthesia in patients undergoing elective colorectal surgery. Anesthesia-controlled time in a ETT/Epidural Anesthesia (EA) group [n = 11; general anesthesia, combined with epidural anesthesia, was maintained by sevoflurane (< 3%) supplemented with a fixed rate of propofol (3 mg/kg/h) under controlled ventilation using the ETT] was compared with that in a LMA/Combined Spinal-Epidural Anesthesia (CSEA) group [n = 10; in combination with spinal-epidural anesthesia, general anesthesia was maintained as the same protocol as the ETT/EA under spontaneous ventilation using the LMA]. Time for airway placement in the LMA/CSEA group was significantly shorter than that in the ETT/EA group. Intervals from the end of surgery until the removal of the airway or the decision to exit the operating room in the LMA/CSEA group were shorter than those in the ETT/EA group. No practical sign of aspiration pneumonia and/or atelectasis was found in patients in either group. Under the circumstance of regional anesthesia being requested for post-surgical pain management, we concluded that the LMA facilitated the emergence from as well as the induction of anesthesia without any practical complication when used for patients in colorectal surgery. PMID- 12587617 TI - A ruptured distal posterior inferior cerebellar aneurysm. Our case and review of the literature. AB - We present a case of ruptured distal posterior inferior cerebellar artery (PICA) aneurysm, and review the literature and discuss the treatment strategy. A 77-year old woman presented with the sudden onset of severe headache, nausea and vomiting. Computed tomography revealed an intraventricular hemorrhage, predominantly in the fourth ventricle and hydrocephalus with a thin subarachinoid hemorrhage (SAH). Angiography revealed an aneurysm arising at the turning point of the vessel, from the telovelotonsillar segment of the right PICA. On the 17th day after the onset, repeated angiography revealed a smaller aneurysm than the one detected on the first day at the same place and with no spasm. On the 22nd day, the aneurysm was proved to be partially thrombosed and was safely clipped via a right lateral suboccipital approach. SAH with a fourth ventricular hemorrhage or an isolated fourth ventricle hemorrhage should raise the suspicion of a distal PICA aneurysm. Aneurysms of the distal PICA have often been reported to arise at a turning point of the artery rather than at a junction of the vessel. It is suggested that the pathogenesis could be hemodynamic stress that has developed due to embryological factors. Distal PICA aneurysms have often gone detected in many previous cases because of thrombosis inside the aneurysms. Thus, particularly in the case of intentionally delayed surgery, we recommend repeated angiography under various conditions to identify how the aneurysm develops just before surgery. PMID- 12587618 TI - Limited value of routine microbiological diagnostics in patients hospitalized for community-acquired pneumonia. AB - Current guidelines recommend microbiological diagnostic procedures as a part of the management of patients hospitalized for community-acquired pneumonia (CAP), but the value of such efforts has been questioned. Patients hospitalized for CAP were studied retrospectively, focusing on the use of aetiological diagnostic methods and their clinical impact. Adult patients, without known human immunodeficiency virus infection, admitted to hospital for CAP during 12 months, were evaluated with regard to the importance of aetiological diagnosis for tailoring antibiotic therapy, antibiotic-associated diarrhoea, Clostridium difficile disease, length of hospital stay and mortality. Of the 605 studied patients, 482 (80%) were subjected to Mycoplasma pneumoniae and/or respiratory virus serology and/or cultures of blood and/or sputum. They had a better prognosis than patients not subjected to microbiological diagnostics (mortality within 3 months was 9% vs 24%, p = 0.001), apparently reflecting differences in general health (e.g. less dementia diagnosis) but not the outcome of diagnostics. A presumptive aetiology was obtained only in 132 of the 482 patients, Streptococcus pneumoniae and M. pneumoniae being the most common agents (in 49 and 36 patients, respectively). Establishing an aetiological diagnosis had no impact on the number of in-hospital changes of therapy, on the proportion of new regimens having a narrower antimicrobial spectrum than the initial one or on the outcome. Therapy was changed to a drug directed specifically against the identified pathogen in only 16 out of these 132 patients and again without any overall improvement in the outcome variables. In a setting with a low frequency of antibiotic-resistant respiratory tract pathogens current routine microbiological diagnostics were found to be of limited value for the clinical management of patients hospitalized for CAP. Improved diagnostics in CAP are urgently needed, as establishing an aetiological diagnosis carries a potential for optimizing the antibiotic therapy. PMID- 12587619 TI - Upper respiratory tract infections in general practice: diagnosis, antibiotic prescribing, duration of symptoms and use of diagnostic tests. AB - A diagnosis/antibiotic prescribing study was performed in 5 counties in Sweden for 1 week in November 2000. As part of this study, the characteristics and clinical management of patients with upper respiratory tract infections (n = 2899) in primary care were analyzed. Almost half of the patients were aged < 15 y and one-fifth of the patients consulted out of hours. Of all patients seeking primary care for upper respiratory tract infections, 56.0% were prescribed an antibiotic. Almost all patients who were given the diagnoses streptococcal tonsillitis, acute otitis media or acute sinusitis were prescribed antibiotics, compared to 10% of patients with common cold or acute pharyngitis. The most frequently prescribed antibiotic was penicillin V (79.2%) and this was even more pronounced out of hours, when the diagnoses otitis media and streptococcal tonsillitis were more frequently used. In patients with common cold and acute pharyngitis, the percentage who received antibiotics increased with increasing length of symptoms and increasing CRP levels. In patients with acute pharyngitis or streptococcal tonsillitis, antibiotics were prescribed less frequently provided streptococcal tests were performed. The management of patients with upper respiratory tract infections in general practice seems to be in good agreement with current Swedish guidelines. However, the study indicates some areas for improvement. The diagnosis of acute sinusitis seems to have been overestimated and used only to justify antibiotic treatment. PMID- 12587620 TI - Management of patients with community-acquired pneumonia treated in hospital in Sweden. AB - To investigate the management of patients with community-acquired pneumonia (CAP) treated in hospital in Sweden, a multicentre retrospective cohort study was performed with medical record review of 982 patients (mean age 63 y) at 17 departments of infectious diseases at hospitals in Sweden. Information on antimicrobial therapy, demographic characteristics, comorbid conditions, physical examination findings, and laboratory and microbiological test results were recorded. Outcome measures were in-hospital mortality and length of hospital stay (LOS). Cultures were obtained from blood in 80% and from sputum in 22% of the patients. A microbiological aetiology was determined for 23% of the patients, with Streptococcus pneumoniae as the dominating agent (9%). The initial antibiotic treatment was mostly given intravenously (78%). Penicillin (50%) or a cephalosporin (30%) was the most common choice. Both of these drugs were usually given as a single agent. The overall mortality was 3.5% and the mean LOS was 6.4 d. Thus, the outcome was favourable despite the empirical antibiotic treatment having a narrow spectrum compared with the broader approach recommended in most recent guidelines on the management of CAP. These findings suggest that a majority of patients who are hospitalized with moderately severe pneumonia can be treated initially with penicillin alone. PMID- 12587621 TI - Randomized, double-blinded, placebo-controlled trial of early administration of recombinant human granulocyte colony-stimulating factor to non-neutropenic preterm newborns between 33 and 36 weeks with presumed sepsis. AB - A randomized, double-blinded, placebo-controlled trial was conducted of early administration of recombinant granulocyte colony-stimulating factor (rGCSF) to 40 non-neutropenic, preterm infants between 33 and 36 weeks of gestational age with the diagnosis of presumed sepsis. The treatment group (n = 20) received 5 microg/kg per day of intravenous rGCSF once daily for 3 d and the control group (n = 20) received the same volume of physiological serum. Immediately before the first dose and on the 4th day, plasma levels of GCSF and tumour necrosis factor alpha (TNF-alpha), absolute neutrophil counts (ANC), immature neutrophil count (INC), immature/total neutrophil (I/T) ratios and platelet counts were determined. At study entry, the plasma GCSF and TNF-alpha levels were similar. On day 4, there was no significant change in GCSF levels in either groups, whereas there was a significant decrease in TNF-alpha levels in the treatment group. ANC and INC of the treatment group also increased significantly. The I/T ratio continued at the same level in the treatment group, but decreased significantly on days 4 and 7 day in the control group. The length of time on the neonatal intensive care unit (NICU) was significantly shorter in the treatment group. In conclusion, early administration of 3 daily doses of rGCSF (5 microg/kg per day) to non-neutropenic, preterm infants who had presumed sepsis increased circulating ANC and INC, decreased plasma TNF-alpha levels and shortened the length of time on the NICU. PMID- 12587622 TI - Single- and multiple-dose pharmacokinetics of oral moxifloxacin and clarithromycin, and concentrations in serum, saliva and faeces. AB - Moxifloxacin and clarithromycin are important antibacterial drugs in the treatment of community-acquired respiratory tract infections. In a double-blind, randomized, 2-period cross-over study the pharmacokinetics of moxifloxacin versus clarithromycin were determined after single and multiple doses in 12 healthy male volunteers. The concentrations of the antibiotics in serum, saliva and faeces were measured by validated high-performance liquid chromatographic methods. In serum, moxifloxacin exhibited a mean peak concentration of 3.1 +/- 0.6 mg/l after a time to peak concentration of 1.67 +/- 0.96 h on day 1, with a significant increase to 3.98 +/- 1.10 mg/l on day 7 (p < 0.05). The area under the curve-12 revealed a highly significant increase from 28.2 + 4.1 mg*h/l on day 1 to 39.5 +/ 6.6 mg*h/l on day 7 (p < 0.01). There were also significant differences in terminal half-life between day 1 and day 7 [10.6 h (range 9.0-12.8) vs 14.9 h (range 12.6-28.1); p < 0.01] and in mean residence time (15.1 +/- 1.9 vs 18.2 +/- 2.4 h; p < 0.01). The concentrations of moxifloxacin in saliva were well equilibrated with serum at a relatively constant saliva-serum ratio of about 0.8. Pharmacokinetic parameters of clarithromycin and its metabolite, 14-hydroxy clarithromycin, were similar to previously published data. Accumulation was found. No serious adverse events were observed with either study drug. PMID- 12587623 TI - Tick-borne encephalitis in north-eastern Poland in 1997-2001: a retrospective study. AB - The aim of this study was to characterize the epidemiology and clinical features of tick-borne encephalitis in north-eastern Poland. Clinical and epidemiological data were analysed of patients hospitalized with the diagnosis in the Department of the Infectious Diseases and Neuroinfections of the Medical University in Bialystok in 1997-2001. Tick-borne encephalitis was diagnosed in 152 patients: 51 (34%) presented with meningitis, 89 (59%) with meningoencephalitis and 12 (8%) with meningoencephalomyelitis. Headache (84%) and fever (81%) were the most common symptoms. Meningeal signs were present in 137 patients (90%). Most common neurological abnormalities were: Oppenheim and Babinski signs (74 patients, 49%), ataxia (37, 24%), impaired consciousness (37, 24%) and pareses (16, 10%). Of patients examined, 146 (96%) had raised pleiocytosis, frequently accompanied by high cerebrospinal fluid protein concentration (90%), raised erythrocyte sedimentation rate (65%), peripheral blood leucocytosis (26%) and increased aminotransferase activity (16%). There was only 1 forest worker among the patients. Tick-borne encephalitis remains common in north-eastern Poland but, possibly because of effective vaccination, it has virtually disappeared among forest employees. The diagnosis appears difficult in some cases, as meningeal signs may not be present and laboratory findings may not be suggestive of a viral infection. PMID- 12587624 TI - A search for optimal criteria in initiating antiretroviral therapy in chronic human immunodeficiency virus infection focusing on CD4 count and HIV RNA. AB - The study objective was to identify optimal starting criteria regarding levels of CD4 cells and human immunodeficiency virus (HIV) RNA at initiation of highly active antiretroviral therapy (HAART) in chronically HIV-infected people. All 162 treatment-naive patients in the centre who were treated for at least 180 d with 2 nucleoside reverse transcriptase inhibitors plus at least 1 protease inhibitor or 1 non-nucleoside reverse transcriptase inhibitor were included. The patients were stratified according to their levels of CD4 cells and HIV RNA at initiation of therapy. Baseline CD4 groups were: group 1: CD4 < 0.1 x 10(9)/l; group 2: CD4 > or = 0.1 and < 0.2 x 10(9)/l; group 3: CD4 > or = 0.2 and < 0.35 x 10(9)/l; and group 4: CD4 > or = 0.35 x 10(9)/l. Two patients died and 38 developed an HIV related disease (Centers for Disease Control category B or C) during the study. The prevalence of HIV-related disease before HAART was significantly increased in groups 1 and 2 compared with groups 3 and 4. The level of HIV RNA was not associated with HIV-related disease either before or after treatment initiation. Subjects in group 1 had an increased risk of HIV-related disease after treatment initiation both in univariate Cox analysis and after adjustment for HIV RNA, gender, mode of transmission and age, compared with group 2 [adjusted risk ratio with 95% confidence interval: 3.76 (1.48-9.61)], group 3 [5.90 (2.07-16.95)] and group 4 [5.05 (1.96-12.90)]. The association between CD4 count and morbidity appeared to be particularly strong for older subjects. In conclusion, this study suggests that in chronically HIV-infected individuals, in most cases HAART can be withheld until the CD4 cell count falls towards 0.2 x 10(9)/l. PMID- 12587625 TI - Giardiasis treatment in Turkish children with a single dose of ornidazole. AB - This study was designed to compare the treatment efficacy of single dose of ornidazole with 5 d treatments of ornidazole and metronidazole in children with giardiasis. 175 children, between 2 and 15 y old, whose stool samples were found to be positive for Giardia lamblia cysts and/or trophozoites by either saline Lugol, formalin-ethyl acetate or trichrome staining, were enrolled in the study. Of these children, 105 were treated with a single dose of ornidazole: 35 with 30 mg/kg, 35 with 25 mg/kg and 35 with 20 mg/kg; 35 were treated with 25 mg/kg per day of ornidazole for 5 d in 2 doses and 35 children were treated with 20 mg/kg per day metronidazole for 7 d in 3 doses. All cases were examined on the 7th, 10th and 14th days after treatment by the same methods; clinical symptoms were also evaluated. Giardia lamblia was eradicated in 34 of 35 (97%), 34 of 35 (97%) and 33 of 35 (94%) patients treated with 30, 25 and 20 mg/kg single doses of ornidazole, respectively. Eradication was achieved in all 35 patients treated with 25 mg/kg per day ornidazole for 5 d and in 31 of 35 (89%) patients treated with metronidazole. There was no statistically significant difference among doses of ornidazole (p > 0.05); however, all ornidazole treatment regimens were significantly more effective than metronidazole treatment (p < 0.05). No important side-effects were detected in any patients and clinical symptoms disappeared in all. Single-dose ornidazole treatment could be considered as a proper and effective alternative method for the treatment of giardiasis in children. PMID- 12587626 TI - Ophthalmia neonatorum caused by group B Streptococcus. AB - A full-term infant was born with a swollen left conjunctival sac. Ophthalmia neonatorum developed within 5 d after birth. Examination of the purulent discharge showed Group B Streptococcus. The CRP level was found to be elevated and parenteral and topical therapy was started. Ophthalmia neonatorum resolved after 7 d of treatment. PMID- 12587627 TI - Activation of latent Lyme borreliosis concurrent with a herpes simplex virus type 1 infection. AB - The case is reported of a 26-y-old woman with latent Lyme borreliosis that was concurrently activated with a herpes simplex virus type 1 infection. Immune suppression by stress may have caused activation of both infections. PMID- 12587628 TI - Stenotrophomonas maltophilia endocarditis. AB - Stenotrophomonas maltophilia is a gram-negative bacillus that is increasingly associated with serious nosocomial infections, especially in immunocompromised patients; however, the occurrence of endocarditis due to this organism is rare. This report describes a case of S. maltophilia endocarditis associated with a central venous catheter. The literature on Stenotrophomonas endocarditis is reviewed. Given the high morbidity and mortality of these infections, early antibiotic therapy utilizing trimethoprim-sulfamethoxazole, along with a second agent and removal of prosthetic devices, is recommended. PMID- 12587629 TI - Port-site tuberculosis: a rare complication following laparoscopic cholecystectomy. AB - The rare complication of port-site tuberculosis following laparoscopic cholecystectomy is reported. In this case, the source of infection was most probably the laparoscope or its accessories. Thus, proper sterilization of the laparoscope and instruments is of utmost importance in preventing infectious complications and ideally autoclaving should be used for this purpose. PMID- 12587631 TI - Multiple tuberculous brain abscesses. AB - Multiple tuberculous brain abscesses are rare. This report presents a female patient with a rapidly deteriorating clinical course. Computed tomographic scanning revealed multiple brain abscesses. Aspiration of one of the lesions yielded pus and positive acid-fast stained bacilli. The pathogenesis of multiple tuberculous brain abscesses is discussed. PMID- 12587630 TI - Nostril infection due to Mycobacterium marinum in an immunocompetent host. AB - Mycobacterium marinum infection developed in the nostril of an immunocompetent host whose only risk factors were infrequent swimming and cleaning of a small fish bowl on a single occasion. The lesion relapsed after 2 surgical excisions but resolved slowly with a 9-month course of trimethoprim/sulfa therapy. PMID- 12587632 TI - Buttock abscess brucellosis. AB - Human brucellosis is a multisystem disease that may present with a broad spectrum of clinical manifestations, and its complications can affect almost all organs and systems with varying incidence. Soft-tissue infection due to Brucella species is a rare complication of brucellosis. This report presents a case of a 30-y-old previously healthy woman whose right buttock abscess culture grew Brucella spp. The patient was treated successfully with abscess drainage and antibiotic therapy. PMID- 12587634 TI - Azithromycin-induced rash in infectious mononucleosis. AB - Antibiotic-induced skin eruption in the setting of infectious mononucleosis is a common and well-documented clinical scenario. The skin lesions are non-specific and the mechanisms causing them are unclear. Several reports have described this entity with different antibiotic classes, mainly penicillins. Only 1 case of azithromycin-induced skin eruption has previously been described in this setting. Herein, we report the second case. PMID- 12587633 TI - Cryptococcal meningitis and sarcoidosis. AB - This report describes a patient with cryptococcal meningitis and newly diagnosed sarcoidosis not taking corticosteroids. Sarcoidosis is an independent risk factor for cryptococcal infection; most patients with sarcoidosis who develop cryptococcal infection are not on immunosuppressive drugs. Cryptococcal meningitis in sarcoid patients often presents clinically with non-specific features, and should be excluded in patients with sarcoidosis and neurological disturbances. PMID- 12587635 TI - Eubacterium bacteremia and colon cancer. AB - Eubacterium bacteremia is rare. We report a senile patient who presented with 3 episodes of bacteremia, caused by Clostridium perfringens, Eubacterium limosum and Escherichia coli, respectively, which led to the diagnosis of adenocarcinoma of the colon. A differential diagnosis of colon cancer should be considered in patients with eubacterium bacteremia. PMID- 12587636 TI - Frequency analysis of the CCR5delta32 mutation in patients with brucellosis. AB - This study investigated the influence of the host's CCR5 genotype in human brucellosis. Genotype analysis for the CCR5delta32 mutation was performed in 185 brucellosis patients and 141 healthy volunteers, all recruited in northern Greece. In the brucellosis-infected patients the CCR5delta32 genotype distribution showed a significant deviation from the Hardy-Weinberg equilibrium (p = 0.030), owing to the presence of 2 individuals homozygous for delta32 among the 185 brucellosis patients compared with none of the 141 controls. Extended studies should be performed to confirm an association between CCR5 genotype and the risk of acquiring brucellosis. PMID- 12587637 TI - Cosmetic breast surgery--is breastfeeding at risk? PMID- 12587638 TI - Is breastfeeding protective against child obesity? AB - Recent studies suggest that breastfeeding reduces the risk of child obesity to a moderate extent. Of 11 studies that examined prevalence of overweight in children older than 3 years of age and that had a sample size of > or = 100 per feeding group, 8 showed a lower risk of overweight in children who had been breastfed, after controlling for potential confounders. The 3 "negative" studies lacked information on the exclusivity of breastfeeding. A dose-response relationship with duration of breastfeeding was observed in some, but not all, of the "positive" studies. Possible mechanisms include learned self-regulation of energy intake, metabolic programming in early life, and residual confounding by parental attributes. If the association is causal, the effect of breastfeeding is probably small compared to other factors that influence child obesity, such as parental overweight. Nonetheless, it may be of public health significance considering the current epidemic of child overweight. PMID- 12587639 TI - Inquiries regarding Kassing's bottle-feeding article. PMID- 12587640 TI - Breastfeeding must be given priority over circumcision. PMID- 12587641 TI - Culture and ethnicity key considerations for increasing breastfeeding rates. PMID- 12587642 TI - Lactation mastitis: a descriptive study of the experience. AB - This descriptive study of 31 women diagnosed with lactation mastitis in the midwestern United States investigated mastitis symptomatology, self-care and treatment, burden of mastitis, and symptom recurrence and complications. Data collected via telephone interviews soon after diagnosis and daily through the seventh day after symptom onset documented that symptom intensity was moderate, peaked on the second day, and lasted on average 3 to 4 days. Symptoms had greater impact on activities of daily living than on breastfeeding. Sources of advice for self-care included lactation consultants, nurses, nurse practitioners, physicians, family, and friends. Continued breastfeeding was the most frequently used of several self-care practices and was rated as highly effective. Follow-up at 2 and 6 weeks established mastitis recurrence as the most common complication. Breastfeeding women need specific information about mastitis causes, symptoms, and self-care strategies to help prevent and treat the condition. PMID- 12587643 TI - Translation and validation of the breastfeeding self-efficacy scale into Spanish: data from a Puerto Rican population. AB - Many new mothers discontinue breastfeeding prematurely due to difficulties encountered rather than maternal choice. Research has shown that a significant predictor of breastfeeding duration is maternal confidence. Using self-efficacy theory as a conceptual framework to measure breastfeeding confidence, the Breastfeeding Self-Efficacy Scale (BSES) was developed and psychometrically tested among English-speaking mothers. The purpose of this methodological study was to translate the BSES into Spanish and determine the psychometric characteristics of the BSES in a sample of 100 Puerto Rican women. The psychometric assessment of the original study was replicated including internal consistency, principal components factor analysis, and comparison between contrasted groups. This study is the first to examine the psychometric characteristics of the Spanish-version BSES administered in-hospital and provide further evidence of the reliability and validity of the instrument. PMID- 12587644 TI - The impact of breast reduction surgery on breastfeeding performance. AB - The impact of reduction mammoplasty surgery on breastfeeding performance was measured comparing a group of 49 Brazilian women who had undergone breast reduction surgery using transposition techniques with 96 controls. As determined by survival analysis, the women who underwent reduction mammoplasty had a significantly shorter time of breastfeeding duration. The prevalence of exclusive breastfeeding at 1 and 4 months was 21% and 4%, respectively, for women with surgery, and 70% and 22%, respectively, for controls (P < .001). The prevalence of any breastfeeding at 1, 6, and 12 months was 58%, 16%, and 10% for women with mammoplasty, and 94%, 58%, and 42% for controls (P < .001). For women with surgery, the median duration of exclusive and any breastfeeding was 5 days and 2 months, respectively, and 3 months and 6 months for controls. The results of this study suggest that breast reduction surgery may have a negative impact on breastfeeding performance. PMID- 12587645 TI - Characteristics of teenage mothers and predictors of breastfeeding initiation in the Michigan WIC Program in 1995. Women, Infants, and Children. AB - Although breast milk is recommended as the optimal source of infant nutrition, breastfeeding initiation is below recommended levels, especially among teenage mothers. Breastfeeding initiation rates among Michigan (US) teenage mothers (12 19 y) were compared by demographics and health behaviors. Multivariate analyses determined which factors were significant independent predictors of breastfeeding initiation among teenage mothers enrolled prenatally in the Michigan Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) program in 1995. Significant predictors independently associated with breastfeeding initiation were race/ethnicity, education, marital status, postpartum anemia status, parity, prenatal trimester of WIC enrollment, and smoking. The strongest predictor of breastfeeding initiation differed for white mothers (positive predictor: education beyond high school [OR = 3.13]) and black mothers (negative predictor: multiparous [OR = 0.25]). Initiation rates for this population of teenage mothers fall below the national average for mothers of all ages and the US Healthy People 2010 goals. Research is needed concerning how breastfeeding support and education can be improved to reach the US national health goals. PMID- 12587646 TI - Feeding preferences and attitudes to breastfeeding and its promotion among teenagers in Northern Ireland. AB - Northern Ireland has one of the lowest breastfeeding initiation rates in the world. Given that attitudes toward infant feeding are formed at an early age and a high rate of teenage pregnancy, it has become necessary to survey attitudes to infant feeding and breastfeeding promotion preferences in teenagers in Northern Ireland. Questionnaires were distributed to teenagers aged 14 to 18 years (n = 419) based in 7 schools selected by type and location throughout Northern Ireland. Attitudes to breastfeeding in public reflected preferred infant-feeding method and were positively influenced by prior exposure to breastfeeding (P = .024). Females were more positive than males both in relation to breastfeeding in public (P = .002) and breastfeeding promotion (P = .003). Recommendations for breastfeeding promotion include specific targeting of young people (both male and female) and enabling contact between teenagers and nursing mothers as much as possible. PMID- 12587647 TI - Breastfeeding after breast augmentation. PMID- 12587649 TI - Printed advice on initiating and maintaining breastfeeding in mid-20th-century Queensland. AB - This article, which is part of a larger historical study of infant-feeding advice received by mothers in Queensland, Australia, identifies and examines print materials that were used by mothers, or that influenced other texts, during the postwar period from 1945 to 1965. The texts are described within the context of their environmental influences and the medical knowledge and research of the time to assess what effect, if any, these latter materials had on the established system of infant-feeding advice. Two innovations that were employed are the use of interviews to identify the most-used texts and the comparison of popular advice books with medical journal material. PMID- 12587648 TI - Peer counselors for breastfeeding mothers in the hospital setting: trials, training, tributes, and tribulations. AB - Boston Medical Center (BMC), an inner-city, Baby-Friendly teaching hospital with approximately 2000 births per year, has employed breastfeeding peer counselors since 1998. The Breastfeeding Center frequently receives requests for information on our peer counselor program. This article reviews program models, training methods, practicalities, benefits, and challenges associated with employing breastfeeding peer counselors in the hospital setting. Specifically, it focuses on 3 different models of peer counseling programs used at BMC: the telephone model, the postpartum model, and the neonatal intensive care model, and it considers the benefits and issues associated with each model. PMID- 12587650 TI - The past and the present. PMID- 12587651 TI - Patient safety concerns grow in Congress. PMID- 12587652 TI - Factor V Leiden with deep venous thrombosis. AB - Factor V Leiden (FVL) is an autosomal co-dominantly inherited Arg506-->Gly substitution of the activated protein C cleavage site affecting 5% of the Caucasian population. FVL results in impaired anticoagulant function without procoagulant modification. Heterozygotes experience a seven-fold increase in thrombotic events, whereas homozygotes may incur a 50 to 100 fold increase. Even though patients are at increased risk for deep venous thrombi, they experience a smaller risk of pulmonary embolism compared to individuals affected by other coagulopathies. PMID- 12587653 TI - Microbiological threats to health in the home. AB - OBJECTIVE: To explore a range of pathogenic microorganisms and their toxins that can cause disease in the home environment through a review of the literature. DESIGN: Review of the literature. PMID- 12587654 TI - Effect of drawing a discard tube on PT and APTT results in healthy adults. AB - OBJECTIVE: The purpose of this study was to compare results obtained for the prothrombin time (PT) and the activated partial thromboplastin time (APTT) using specimens drawn with and without a discard tube in healthy adults. DESIGN: A specimen of blood in a 3.2% sodium citrate, 5.0 mL tube was drawn from one arm with a discard tube and from the other arm without a discard tube on 35 healthy adults. A PT and APTT were performed on each specimen using a fibrometer. SETTING: The PT and APTT tests were all performed in the student laboratory of the Clinical Laboratory Science Program at East Carolina University. PATIENTS OR OTHER PARTICIPANTS: Study subjects induded technician, technologist, and phlebotomist employees of the clinical laboratory at Pitt County Memorial Hospital, and faculty and junior and senior students in the Clinical Laboratory Science Program at East Carolina University, Greenville, North Carolina. All participants signed an informed consent prior to venipuncture. INTERVENTIONS: Two coagulation specimens were drawn from each subject. Specimens in a discard (no anticoagulant) tube and then an anticoagulated tube were drawn from one arm, and a specimen in a single anticoagulant tube was drawn from the other arm. The PT and APTT were performed using standardized procedures with Pacific Hemostasis reagents and controls and a BBL FibroSystem fibrometer. MAIN OUTCOME MEASURES: The degree of difference between PT results and between APTT results for specimens drawn with and without a discard tube. RESULTS: Based on paired t-test analyses, no difference in mean PT results and no difference in mean APTT results were found between specimens drawn with a discard tube and those drawn without a discard tube at an alpha of 0.05. Paired-samples correlation coefficients were significant for both the PT and the APTT at an alpha of 0.05, showing precision between results with and without a discard tube for both coagulation tests. CONCLUSIONS: Relative to sampling from a population of healthy adults, drawing a discard tube before a sodium citrate tube for coagulation testing appears to make an insignificant difference. Replication of these results with patients receiving anticoagulant therapy and/or patients with abnormal coagulation results, would offer cost savings by justifying elimination of discard tubes for blood draws for coagulation testing only. Such a change in protocol would also reduce the likelihood of nosocomial blood loss in vulnerable patient populations. PMID- 12587655 TI - Motivations for condom use and nonuse. AB - This exploratory study investigates the motivations for condom use and nonuse among a sample of drug users and nonusers. Participants who reported condom use in the previous 30 days rated various reasons for using a condom the last time they had had sex (79 participants). Those who reported not using a condom rated reasons for not using a condom the last time they had had sex (147 participants). Factor analyses of these responses were used to summarize reasons for condom use and nonuse. It was found that a peer norms motivation and a relationship motivation (caring) supported both condom use and nonuse. A moral norms motivation supported condom use, but a motivation that denied moral norms supported nonuse. A self interest motivation of personal pleasure supported nonuse. No strictly self-interest motivation was identified for condom use: self protection was associated with protection of the partner. PMID- 12587656 TI - Dengue fever in the Western Hemisphere. AB - Dengue virus, an arthropod-borne viral agent, causes two distinct diseases: classic dengue fever (DF) and dengue hemorrhagic fever (DHF). There are four dengue virus serotypes: DEN-1, DEN-2, DEN-3, and DEN-4. Although infection with dengue stimulates immunologic response to a serotype, there is no cross-immunity conferred. Hence, a person can potentially be infected with each serotype during his or her lifetime. An infected female Aedes mosquito transmits the virus from person to person while feeding. The disease, now endemic in more than 100 countries in Africa, the Americas, the Eastern Mediterranean, Southeast Asia, and Western Pacific, is spreading to new areas and causing explosive outbreaks. Because of the major impact on lives and local economies epidemics produce, rapid detection of dengue infection has become an important public health research issue. Recently developed serological procedures to detect dengue infections have shown great potential for field use. PMID- 12587657 TI - Hantavirus. AB - Sin Nombre virus (SNV), an emerging pathogen in the United States, was identified in 1993. This hantavirus, a member of the Bunyaviridae family of RNA viruses, is transmitted by its reservoir host Peromyscus maniculatus, the deer mouse. Transmission is by inhalation of aerosolized feces, urine, or saliva from the infected mice. The illness that pursues, hantavirus pulmonary syndrome (HPS), is characterized initially by mild flu-like symptoms, followed by rapid progression to respiratory distress. There is no established therapeutic regimen and treatment is only supportive. Preventive methods include attempts to minimize contact with the rodents since elimination of the virus is not realistic. PMID- 12587658 TI - West Nile virus: an emerging virus in North America. AB - West Nile virus is an emerging virus that first appeared in North America during the summer of 1999 in New York City. There were seven deaths associated with this event. Surveillance reports indicate that the virus had been spreading south and west and in 2002, had been reported in 42 states and the District of Columbia. As of September 2002, there were 2121 total human cases reported, induding 104 deaths. The fatality rate for the West Nile virus is very low and the majority of individuals will have no clinical symptoms; however, individuals at most risk for more serious form of the disease are the elderly, the immunocompromised, and young individuals. The virus is spread by certain mosquito species and certain populations of birds serve as the reservoir hosts. Because person-to-person transmission does not occur, humans are therefore considered dead-end hosts. Confirmation of cases West Nile virus infections in humans are determined based on clinical and laboratory findings. PMID- 12587659 TI - Turnover of myelin lipids in aging brain. AB - Turnover rates of myelin membrane components in mouse brains were determined by a method using stable isotope-labeling and mass spectrometry. The half-replacement times based on incorporation rates of newly synthesized molecules for young adult mice were 359 days for cholesterol, 20 days for phosphatidylcholine, 25 days for phosphatidylethanolamine, 94 days for cerebroside and 102 days for ganglioside GM1. The turnover rates of half-lives of myelin components were calculated from the decay curves of initially labeled molecules, and they were about the same as the half-replacement times. Individual components were thus revealed to be metabolized at different rates, and their turnover rates were differently affected by aging. As was observed with phospholipids, myelin pools appeared to be compartmentalized into rapidly and slowly exchanging pools. The turnover rates of cerebroside and GM1 decreased between the young and adult periods and slightly increased in senescence. The latter phenomenon may indicate an enhanced myelin turnover in senescence. The present study reveals the dynamic aspects of myelin membrane turnover during the life span of mouse. PMID- 12587660 TI - p53-dependent cell death signaling in neurons. AB - The p53 tumor suppressor gene is a sequence-specific transcription factor that activates the expression of genes engaged in promoting growth arrest or cell death in response to multiple forms of cellular stress. p53 expression is elevated in damaged neurons in acute models of injury such as ischemia and epilepsy and in brain tissue samples derived from animal models and patients with chronic neurodegenerative diseases. p53 deficiency or p53 inhibition protects neurons from a wide variety of acute toxic insults. Signal transduction pathways associated with p53-induced neuronal cell death are being characterized, suggesting that intervention may prove effective in maintaining neuronal viability and restoring function following neural injury and disease. PMID- 12587661 TI - Spatiotemporal distribution of the insulin-like growth factor receptor in the rat olfactory bulb. AB - Insulin-like growth factor I (IGF-I) and its receptor (IGF-IR) are involved in growth of neurons. In the rat olfactory epithelium, we previously showed IGF-IR immunostaining in subsets of olfactory receptor neurons. We now report that IGF IR staining was heaviest in the olfactory nerve layer of the rat olfactory bulb at embryonic days 18, and 19 and postnatal day 1, with labeling of protoglomeruli. In the adult, only a few glomeruli were IGF-IR-positive, some of which were unusually small and strongly labeled. Some IGF-IR-positive fibers penetrated deeper into the external plexiform layer, even in adults. In developing tissues, IGF-IR staining co-localized with that for olfactory marker protein and growth associated protein GAP-43, but to a lesser extent with synaptophysin. In the adult, IGF-IR-positive fibers were compartmentalized within glomeruli. IGF-I may play a role in glomerular synaptogenesis and/or plasticity, possibly contributing to development of coding patterns for odor detection or identification. PMID- 12587662 TI - Inhibitors of DNA binding in neural cell proliferation and differentiation. AB - Id proteins function as negative regulators of bHLH transcription factors by disrupting the homo- and/or hetero-dimerization of bHLH-bHLH transcription factors. Recent data from in vitro and in vivo studies have revealed the complex biological functions of Id proteins in the regulation of cell differentiation, the cell cycle, and cell survival. Several advances in the understanding of Id regulated neurogenesis have been made. Basically, Id proteins are positive regulators of neural cell proliferation, are required for neural cell cycle progression, and also play a role in the timing of oligodendroglial differentiation. Here we summarize recent findings regarding the regulation of Id proteins in neural cells and discuss the possible mechanisms of Id-regulated neurogenesis. PMID- 12587663 TI - Role of the JAK/STAT signal transduction pathway in the regulation of gene expression in CNS. AB - Over the last 20 years the JAK/STAT signal transduction pathway has been extensively studied. An enormous amount of data on different cell signal transduction pathways is now available. The JAK/STAT signal transduction pathway is one of the intracellular signaling pathways activated by cytokines and growth factors that was first studied in the hematopoietic system, but recent data demonstrate that this signal transduction is also greatly utilized by other systems. The JAK/STAT pathway is a signaling cascade that links the activation of specific cell membrane receptors to nuclear gene expression. This review is focused on the role of JAK/STAT signal transduction pathway activation in the central nervous system (CNS). PMID- 12587664 TI - The involvement of lipid activators of protein kinase C in the induction of ZIF268 in PC12 cells exposed to lead. AB - Lead (Pb) induces the expression of immediate early genes (IEG) in PC12 cells by a mechanism that involves protein kinase C (PKC). To define the mechanisms, the involvement of two commonly observed lipid activators of PKC, diacylglycerols, and phosphatidylinositols, were examined. A dose-dependent increase in the expression of the IEG zif268 was observed in PC12 cells exposed to Pb. The PKC inhibitor Ro-31-8220 blocked the induction. An increase in levels of diacylglycerols was observed in PC12 cells exposed to Pb, but the increase was inhibited by Ro-31-8220. The phosphatidylinositol 3-kinase inhibitor Wortmannin, but not the inhibitor LY 294002, blocked the induction zif268 in Pb-exposed cells. Small increases in phosphatidylinositol 3-kinase activity were observed after exposure to Pb. In summary, diacylglycerols are elevated in PC12 cells exposed to Pb by a mechanism that requires PKC. It is possible that diacylglycerols contribute to the induction of zif268 by Pb by sustaining PKC activation. PMID- 12587665 TI - Association of seven polymorphisms of the D2 dopamine receptor gene with brain receptor-binding characteristics. AB - Association of alleles at the Taq1 A, Taq1 B, intron 6, Taq1 D, exon 7, exon 8, and promoter-141C sites of the D2 dopamine receptor gene with D2 dopamine receptor binding characteristics in the caudate nucleus of Caucasian alcoholic and nonalcoholic subjects was determined. For the Taq1 D, exon 7, exon 8, and promoter- 141C sites there were no significant allelic differences in Bmax (number of binding sites) or Kd (binding affinity) of the D2 dopamine receptors. However, subjects having the minor alleles at the Taq1 A, Taq1 B, and intron 6 sites had significantly lower Bmax than subjects not having them. None of these three polymorphisms had any significant effect on Kd. Highly significant linkage disequilibria were observed among the Taq1 A, Taq1 B, and intron 6 polymorphic sites, but linkage disequilibria between these three sites and each of the Taq1 D, exon 7, exon 8, and promoter-141C sites were of lesser or of no significance. Taken together, these findings suggest that the Taq1 A, Taq1 B, and intron 6 polymorphisms, but not the Taq1 D, exon 7, exon 8, and promoter-141C polymorphisms, are in linkage disequilibrium with a functional allelic variant that affects D2 dopamine receptor expression. PMID- 12587666 TI - Increased fibrillar beta-amyloid in response to human clq injections into hippocampus and cortex of APP+PS1 transgenic mice. AB - Human C1q when injected directly into hippocampus and cortex of doubly transgenic APP+PS1 mice results in the increase of Congo red-positive fibrillar deposits. Although there was no significant change in overall area stained for Abeta total, qualitatively it appeared that there was less diffuse Abeta in C1q-treated mice versus vehicle. There was no apparent change in astroglial or microglial activation caused by injection of C1q with respect to vehicle injections. These effects of C1q were only found in 50% BUB/BnJ mice, a strain with higher serum complement activity than other mouse lines. These in vivo data were consistent with the effects of C1q to increase fibrillogenesis of Abeta in vitro. In conclusion, complement protein C1q, believed to be involved in the pathogenesis of Alzheimer's disease in humans, can cause increased fibrillogenesis in the APP+PS1 mouse model of amyloid deposition. PMID- 12587668 TI - Morphological changes of myelin sheaths in rats intracranially injected with apotransferrin. AB - Previous findings from our laboratories indicate that the intracranial injection of apotransferrin (aTf) in neonatal rats produces an accelerated oligodendrocyte maturation and an enhanced production and deposition of myelin membranes in the brain. To evaluate the anatomical distribution and the morphological characteristics of the myelin in these rats, we analyzed the optic nerves, cerebellum, and selected areas of brain sections from aTf-treated and control rats by both light and electron microscopy. Microscopic identification of myelin using a specific staining procedure, showed that in aTf-injected rats, in coincidence with previous biochemical studies, there was an increased deposition of myelin in selected areas of the nervous system. Qualitative and quantitative analysis of electron micrographs from areas showing increased myelinaton, such as the optic nerves and the corpus callosum, showed that among other changes, the intracranial treatment with aTf produces ultrastructural evidences of myelin decompaction, consisting of an enlargement in the distance between adjacent major dense lines, a decreased density of the intraperiod line, and an increased electron density of the major dense line, accompanied by a significant increase in its width. The intracranial administration of aTf induces an increased deposition of myelin by oligodeudroglial cells (OLGc), and these myelin membranes, in spite of the changes in composition and in morphology, appear to function normally. Apotransferrin can be considered as a differentiation factor that could be used to stimulate remyelination in cases in which myelin has been destroyed by various pathological processes. PMID- 12587667 TI - Response of chemokine antagonists to inflammation in injured spinal cord. AB - Inflammation is a primary reaction to infection, allergic disorders, autoimmune diseases, and mechanical injury. The goal of an inflammatory response is to rapidly respond to noxious stimuli, such as trauma or pathogen, with a controlled amplification of cellular activation to eliminate, control, or wall off the triggering agent. Although the inflammatory response is necessary for resolution of the pathogenic event, by stander or collateral tissue damage is caused by the toxic nature of many of its by-products. It is characterized by the infiltration of leukocytes into the affected area. Chemokines and their receptors play an essential role as mediators of leukocyte infiltration. In most cases this response is so vigorous that its control, especially in the central nervous system, would inhibit recovery. The benefits of anti-inflammatory therapy based on interference with the chemokine system has been established in animal models and is being pursued with chemokine antibodies and receptor antagonists. Prolonged treatment with a broad-spectrum chemokine antagonist, vMIPII, has been shown to reduce the rate of infiltration of monocytes into injured rat spinal cord and promote survival. PMID- 12587669 TI - Hyperoxia increases AP-1 DNA binding in rat brain. AB - Oxidative stress appears to contribute to neurodegenerative outcomes after ischemia, hypoxia, and hyperoxia. The AP-1 transcription factor is made up of a family of regulatory proteins that can be activated by oxidative stress. In the present study, we examined AP-1 DNA binding activity in terms of specific participating AP-1 proteins in rat brain after hyperoxia. Male Sprague-Dawley rats were exposed to 100% oxygen under isobaric conditions over time. The AP-1 DNA binding activity present in the rat hippocampus and basal forebrain was characterized by electrophoretic mobility shift analysis (EMSA) and the participating AP-1 proteins identified by immunodepletion/supershift and Western blotting analyses. The Fos and Jun proteins were localized by immunohistochemistry to hippocampus. There were significant increases in AP-1 DNA binding in both hippocampus and basal forebrain after hyperoxia. There was also a significant increase in c-Jun protein levels and the proportion of c-Jun present in AP-1 DNA binding complexes in hippocampal nuclei after hyperoxia. These results suggest that AP-1 activation via c-Jun binding to DNA is an important component of brain responses to oxidative stress. PMID- 12587670 TI - Initiation of sodium channel clustering at the node of Ranvier in the mouse optic nerve. AB - Ion fluxes in mammalian myelinated axons are restricted to the nodes of Ranvier, where, in particular, voltage-gated Na+ channels are clustered at a high density. The node of Ranvier is separated from the internode by two distinct domains of the axolemma, the paranode and the juxtaparanode. Each axonal domain is characterized by the presence of a specific protein complex. Although oligodendrocytes and/or myelin membranes are believed to play some instructive roles in the organization of axonal domains, the mechanisms leading to their localized distribution are not well understood. In this paper we focused on the involvement of myelin sheaths in this domain organization and examined the distribution of axonal components in the optic nerves of wild type, hypomyelinatingjimpy mice and demyelinating PLP transgenic mice. The results showed that the clustering of Na+ channels does not require junction-like structures to be formed between the glial processes and axons, but requires mature oligodendrocytes to be present in close vicinity. PMID- 12587671 TI - Intracellular signaling by the neural cell adhesion molecule. AB - Cell adhesion molecules are known to play far more complex roles than mechanically attaching one cell to an adjacent cell or to components of the extracellular matrix. Thus, important roles for cell adhesion molecules in the regulation of intracellular signaling pathways have been revealed. In this review, we discuss the present knowledge about signaling pathways activated upon homophilic binding of the neural cell adhesion molecule (NCAM). Homophilic NCAM binding leads to activation of a signal transduction pathway involving Ca2+ through activation of the fibroblast growth factor receptor, and to activation of the mitogen-activated protein kinase pathway. In addition, cyclic adenosine monophosphate and protein kinase A are involved in NCAM-mediated signaling. Among these pathways the possibility exists of cross talk or convergence, of which different possible mediators have been suggested. Finally, several downstream effector molecules leading to NCAM-mediated cellular endpoints have been demonstrated, including transcription factors and regulators of the cytoskeleton. PMID- 12587672 TI - Protection of mature oligodendrocytes by inhibitors of caspases and calpains. AB - Mature mouse oligodendrocytes (OLs) are susceptible to death in demyelinating diseases such as multiple sclerosis and in brain injury following neurotrauma, ischemia, or stroke. To understand mechanisms leading to death of mature OLs and develop strategies for protection, we utilized cultures of mature mouse OLs to investigate the role of caspases and calpains in OL cell death mediated by different mechanisms. The agents used were (i) staurosporine, which induces apoptotic death via inhibition of protein kinases; (ii) kainate, which activates non-NMDA glutamate receptors; (iii) thapsigargin, which releases intracellular calcium stores; and (iv) SNAP, which releases active NO species and causes necrotic cell death. Inhibitors blocking primary effector caspases (including caspase 3), the FAS (death receptor)-mediated initiator caspases (including caspase 8), and stress-induced caspases (including caspase 9), were tested for their protective effects. Inhibition of caspases 3, 8, and 9 each robustly protected OLs following insult with staurosporine, thapsigargin, or kainate when added at optimal times. The time of addition of the inhibitors for maximal protection varied with the agent, from 1 h of preincubation before addition of staurosporine to 6 h after addition of kainate. Much less protection was seen for the NO generator SNAP under any condition. The role of calcium in OL death in each model was investigated by chelating extracellular Ca++ with EGTA, and by inhibiting the Ca++-activated calpain proteases. Calcium chelation did not protect against staurosporine, but decreased OL death initiated by kainate, thapsigargin, or NO. The calpain inhibitors PD150606 and calpain inhibitor I protected from cell death initiated by staurosporine, kainate, and thapsigargin, but not from cell death initiated by the NO donor SNAP. PMID- 12587673 TI - Lead-induced endoplasmic reticulum (ER) stress responses in the nervous system. AB - Lead (Pb) poisoning continues to be a significant health risk because of its pervasiveness in the environment, its known neurotoxic effects in children, and potential endogenous exposure from Pb deposited in bone. New information about mechanisms by which Pb enters cells and its organelle targets within cells are briefly reviewed. Toxic effects of Pb on the endoplasmic reticulum (ER) are considered in detail, based on recent evidence that Pb induces the expression of the gene for 78-kD glucose-regulated protein (GRP78) and other ER stress genes. GRP78 is a molecular chaperone that binds transiently to proteins traversing through the ER and facilitates their folding, assembly, and transport. Models are presented for the induction of ER stress by Pb in astrocytes, the major cell type of the central nervous system, in which Pb accumulates. A key feature of the models is disruption of GRP78 function by direct Pb binding. Possible pathways by which Pb-bound GRP78 stimulates the unfolded protein response (UPR) in the ER are discussed, specifically transduction by IRE1/ATF6 and/or IRE1/JNK. The effect of Pb binding to GRP78 in the ER is expected to be a key component for understanding mechanisms of Pb-induced ER stress gene expression. PMID- 12587674 TI - Muscle reinnervation and IGF-I synthesis are affected by exposure to heparin: an effect partially antagonized by anti-growth hormone-releasing hormone. AB - Sciatic nerve crush was performed in 2-day-old rats, then reinnervation of the extensor digitorum longus muscle, motor neuron survival, and muscle IGF-I production were monitored. In saline-treated rats, the extent of reinnervation was around 50% and the number of EDL reinnervating motor neurons was significantly reduced. In heparin-treated rats the extent of muscle reinnervation, the recovery of nerve-evoked muscle twitch tension, and the number of motor neurons reinnervating the extensor digitorum longus muscle were greatly enhanced compared to saline-treated rats. In addition, treatment with heparin increased markedly insulin-like growth factor-I levels in denervated muscles. The concomitant exposure to anti-growth hormone releasing hormone partially abolished the stimulatory action of heparin on muscle reinnervation and prevented the increase of insulin-like growth factor-I muscle levels. PMID- 12587675 TI - Quantitative determination of the hydrolysis products of nitrogen mustards in human urine by liquid chromatography-electrospray ionization tandem mass spectrometry. AB - Nitrogen mustards are a public health concern because of their extreme vesicant properties and the possible exposure of workers during the destruction of chemical stockpiles. A sensitive, rapid, accurate, and precise analysis for the quantitation of ultratrace levels of N-ethyldiethanolamine (EDEA) and N methyldiethanolamine (MDEA) in human urine as a means of assessing recent exposure to the nitrogen mustards bis(2-chloroethyl)ethylamine and bis(2 chloroethyl)methylamine, respectively, was developed. The method was based on solid-phase extraction, followed by analysis of the urine extract using isotope dilution high-performance liquid chromatography-mass spectrometry with TurbolonSpray ionization and multiple-reaction monitoring. The method limits of detection were 0.41 ng/mL for EDEA and 0.96 ng/mL for MDEA in 1 mL of urine with coefficients of variation < 10% for both compounds. PMID- 12587676 TI - A general unknown screening procedure for drugs and toxic compounds in serum using liquid chromatography-electrospray-single quadrupole mass spectrometry. AB - A complete general unknown screening procedure was developed using liquid chromatography-mass spectrometry (LC-MS), a coupling that can increase the range of compounds amenable to MS. Sample preparation was by solid-phase extraction on a mixed-mode support in parallel with serum deproteination in order to recover the most hydrophilic compounds. Chromatography employed a reversed-phase narrow bore column (150 x 1-mm i.d.) and a 50-min gradient elution at low flow-rate (50 microL/min), compatible with the electrospray source used without splitting nor heating. The single quadrupole LC-MS instrument used was operated in the 100 to 1100 mu mass range in both the positive and negative modes, with two different, alternated collision-induced dissociation voltages in the source, in order to obtain the molecular or pseudo-molecular ions as well as fragments for the compounds analyzed. The addition of spectra obtained at low and high fragmentation voltages gave reconstructed spectra for each polarity, representing library entries. Finally, a program was created in order to detect the peaks of interest in the chromatographic noise using a very efficient signal processing algorithm, compute their relative retention time with respect to the internal standard (glafenine), draw their reconstructed spectra, search them in the libraries, and edit a report. PMID- 12587677 TI - Direct screening of urine for MDMA and MDA by liquid chromatography-tandem mass spectrometry. AB - There is a limit in the number of substances detected by commercially available reagents. It is therefore important to have other, complementary techniques. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) may offer one possibility. An LC-MS-MS procedure based on the detection of positive ions after atmospheric pressure chemical ionization (APCI) was developed for direct measurement of 3,4-methylenedioxymethamphetamine (MDMA) and 3,4 methylenedioxyamphetamine in urine. It was compared with Online Amphetamines. The LC-MS-MS methodology showed improved sensitivity (1 00-ng/mL cutoff and specificity with a coefficient of variation of 10%. It was compared to the immunochemical analysis using 1000 clinical patient urine samples. The positive samples were confirmed by gas chromatography-mass spectrometry. The LC-MS-MS method detected almost four times as many samples positive for MDMA compared to the immunochemical method, with no false positives and one false negative. Our study suggests that LC-MS-MS offers an alternative to immunochemical methods in drug of abuse screening. PMID- 12587678 TI - Interindividual dose/concentration relationship for methadone in hair. AB - Hair samples were collected from 60 patients receiving long-term methadone maintenance: 50 were taking the drug orally and 10 were receiving the drug by intravenous injection. The amount of methadone present in the hair samples was measured using methanolic extraction, derivatization of the extracts with MTBSTFA, followed by electron impact gas chromatography-mass spectrometry operating in selected ion monitoring mode. The limit of quantitation for the assay was 0.4 ng/mg hair. The dose/concentration relationship for methadone in hair was investigated. No interindividual correlation between prescribed dose and concentration of methadone in hair was observed. PMID- 12587679 TI - A selective and sensitive method for quantitation of lysergic acid diethylamide (LSD) in whole blood by gas chromatography-ion trap tandem mass spectrometry. AB - A gas chromatography-ion trap tandem mass spectrometry (GC-ion trap MS-MS) method for detection and quantitation of LSD in whole blood is presented. The sample preparation process, including a solid-phase extraction step with Bond Elut cartridges, was performed with 2 mL of whole blood. Eight microliters of the purified extract was injected with a cold on-column injection method. Positive chemical ionization was performed using acetonitrile as reagent gas; LSD was detected in the MS-MS mode. The chromatograms obtained from blood extracts showed the great selectivity of the method. GC-MS quantitation was performed using lysergic acid methylpropylamide as the internal standard. The response of the MS was linear for concentrations ranging from 0.02 ng/mL (detection threshold) to 10.0 ng/mL. Several parameters such as the choice of the capillary column, the choice of the internal standard and that of the ionization mode (positive CI vs. EI) were rationalized. Decomposition pathways under both ionization modes were studied. Within-day and between-day stability were evaluated. PMID- 12587680 TI - LC-MS analysis of serotonergic drugs. AB - Separating drugs which are both polar and basic has long been difficult because of the limited operating pH range of conventional HPLC columns. This paper describes a liquid chromatographic method capable of being used with either diode array or mass spectrometric detection for the identification and quantitation of 10 antidepressant and 2 antipsychotic drugs, all of which have serotonergic activity. In developing the method, the effects of varying buffers and mobile phase pH and of adding modifying agents on resolution and capacity factors were investigated. The organic buffers ammonia, glycine, and triethylamine were each used in a mobile phase made up of 32.5% buffer/67.5% methanol (v/v) at a pH of 10.0. Additionally, four different concentrations each of tetrahydrofuran and acetonitrile were added to investigate the effect of a modifying agent on resolution and retention. In general, decreasing mobile phase pH reduced retention times and decreased resolution. Adding tetrahydrofuran in place of the same amount of methanol tended to decrease retention times, and adding acetonitrile tended to slightly increase retention times. However, addition of both marginally improved resolution. This method has been used to satisfactorily analyze brain, blood, liver, urine, vitreous fluid, and stomach contents in subjects known to have used these drugs. PMID- 12587681 TI - Determination of chromate adulteration of human urine by automated colorimetric and capillary ion electrophoretic analyses. AB - Various chemicals can be added to urine specimens collected for drug analysis to abnormally elevate ionic concentrations and/or interfere with either immunoassay urine drug-screening procedures or gas chromatographic-mass spectrometric confirmation techniques. One such adulterant, "Urine Luck" (formula 5.3), has been identified in our previous research to contain potassium dichromate. Screening of suspected adulterated specimens and confirmation of the adulterant are important for forensic drug screening. The application and comparison of automated colorimetric and capillary ion electrophoretic techniques for the detection, confirmation, and quantitation of chromate adulteration of urine specimens were the purpose of this investigation. Thirty-six urine specimens suspected of adulteration were analyzed for chromate by colorimetric analysis with diphenylcarbazide. Duplicate aliquots were analyzed for chromate by capillary ion electrophoresis. Results of the colorimetric chromate analyses revealed a mean chromate concentration of 929 microg/mL with a standard error of 177 microg/mL and a range of 30 to 5634 microg/mL. Results of the capillary ion electrophoresis chromate analyses revealed a mean chromate concentration of 1009 microg/mL with a standard error of 218 microg/mL and a range of 20 to 7501 microg/mL. The correlation coefficient between the capillary ion electrophoretic and colorimetric chromate results was r = 0.9669. Application of the automated diphenylcarbazide colorimetric technique provides rapid determination of chromate adulteration of a urine specimen. Capillary ion electrophoresis offers a separation technique to confirm the presence of chromate in suspected adulterated specimens. The excellent correlation between these methods substantiates their application to forensic testing as screening and/or confirmation techniques. PMID- 12587682 TI - A study of urinary endogenous gamma-hydroxybutyrate (GHB) levels. AB - In recent years, the use of gamma-hydroxybutyrate (GHB), as a recreational drug has prompted forensic toxicology laboratories to incorporate the analysis for GHB into their routine screening procedures. GHB, being a natural occurring constituent of the human body, presents a challenge for forensic toxicologists in that endogenous levels and exogenous levels of GHB need to be differentiated in case samples. This study was designed to determine typical urinary endogenous levels of GHB in humans based on the analysis of urine samples voluntarily provided by 55 male and female subjects ranging in age from 6 to 59 years. All samples were initially screened for the presence of GHB utilizing a hydrolysis method designed to quantitatively convert the GHB in urine samples to gamma butyrolactone (GBL) followed by the liquid-liquid extraction and analysis of any GBL present by gas chromatography-mass spectrometry (GC-MS). As a confirmation test, samples were then extracted by a solid-phase extraction technique, derivatized to GHB di-TMS, and analyzed by GC-MS. The median concentration determined for the 55 subjects was 1.3 mg/mL (mean = 1.65 microg/mL, range 0.9 microg/mL to 3.5 microg/mL, standard deviation 0.68 microg/mL). The results of this study confirm the previously suggested cutoff of 10 microg/mL for routine forensic analyses. PMID- 12587683 TI - An improved HPLC analysis of the metabolite furoic acid in the urine of workers occupationally exposed to furfural. AB - An improved high-performance liquid chromatographic (HPLC) method for the analysis of the metabolite furoic acid in the urine of workers occupationally exposed to furfural is described. The procedure involved an alkaline hydrolysis step followed by solvent extraction using ethyl acetate. HPLC analysis used an acidic acetonitrile/water mobile phase with a C18 column and ultraviolet detection. The overall relative recovery of furoic acid in urine was found to be 98.8% with a relative standard deviation of 9.7%. The limit of quantitation was determined to be 0.01 mmol/L. PMID- 12587684 TI - Analysis of flecainide and two metabolites in biological specimens by HPLC: application to a fatal intoxication. AB - A few days after her admittance to a hospital for a suicide attempt with benzodiazepines, a 15-year-old girl was found dead in bed. At autopsy, no specific anatomo-pathologic cause of death was identified. Systematic toxicological analysis (HPLC-DAD, GC-NPD, and GC-MS) of postmortem blood and urine revealed the presence of high concentrations of flecainide and its two major metabolites. Flecainide is a class IC anti-arrhythmic drug causing a decreased intracardiac conduction velocity in all parts of the heart. To identify and quantitate flecainide together with its metabolites in blood, urine, and other toxicologically relevant matrices, a new method was developed using high performance liquid chromatography with diode-array detection. All compounds were separated on a Hypersil BDS phenyl column using water, methanol, and 1.5M ammonium acetate in a gradient system. Chromatographic analysis was preceded by an optimized solid-phase extraction procedure on RP-C18 extraction columns. The flecainide concentrations in blood and urine were 18.73 and 28.3 mg/L, respectively, and the metabolites were detected only in urine at the following concentrations: 9.4 mg/L for meta-O-dealkylated flecainide and 8.59 mg/L for meta O-dealkylated flecainide lactam. Based on these results, it was concluded that the suicide was consistent with an overdose of this anti-arrhythmic drug. PMID- 12587685 TI - Urinary concentrations of morphine and codeine after consumption of poppy seeds. AB - A quantitative analysis of morphine and codeine in human urine was performed after oral intake of cakes containing commercially available poppy seeds in order to estimate the possibility of positive doping results. Therefore, eight products from different manufacturers (poppy seeds or baking mixtures) and origin were obtained and analyzed by gas chromatography-mass spectrometry for the presence of the alkaloids. One selected batch of poppy seeds was used as an ingredient in a typical cake and was the object of an excretion study with nine volunteers. After application, several urine specimens contained morphine with concentrations higher than 1 microg/mL, and peak values of approximately 10.0 microg/mL were detected. Because the International Olympic Committee set a cutoff limit for morphine at 1 microg/mL, high-performance athletes could possibly test positive in doping control after consumption of products containing poppy seeds. PMID- 12587686 TI - Bromelain reduces mild acute knee pain and improves well-being in a dose dependent fashion in an open study of otherwise healthy adults. AB - There is preliminary clinical evidence to support the contention that the anti inflammatory and analgesic properties of bromelain help to reduce symptoms of osteo- and rheumatoid arthritis. However, there have been no controlled studies of its effects on joint health in healthy subjects who lack such diagnosis. The current study investigated the effects of bromelain on mild acute knee pain of less than 3 months duration in otherwise healthy adults. The study was an open, dose-ranging postal study in volunteers who had been recruited through newspaper and magazine articles. Two validated questionnaires (WOMAC knee health Index and the Psychological Well-Being Index) were completed at baseline and after one month's intervention with bromelain, randomly allocated to volunteers as either 200 mg or 400 mg per day. Seventy seven subjects completed the study. In both treatment groups, all WOMAC symptom dimension scores were significantly reduced compared with baseline, with reductions in the final battery (total symptom score) of 41 and 59% (P = 0.0001 and <0.0001) in the low and high dose groups respectively. In addition, improvements in total symptom score (P = 0.036) and the stiffness (P = 0.026) and physical function (P = 0.021) dimensions were significantly greater in the high-dose (400 mg per day) compared with the low dose group. Compared to baseline, overall psychological well-being was significantly improved in both groups after treatment (P = 0.015 and P = 0.0003 in the low and high dose groups respectively), and again, a significant dose response relationship was observed. We conclude that bromelain may be effective in ameliorating physical symptoms and improving general well-being in otherwise healthy adults suffering from mild knee pain in a dose-dependant manner. Double blind, placebo-controlled studies are now warranted to confirm these results. PMID- 12587687 TI - Choleretic activity and biliary elimination of lipids and bile acids induced by an artichoke leaf extract in rats. AB - The therapeutic properties of artichoke (Cynara scolymus L.) preparations have been known since ancient times. The traditional use of artichoke leaf extract (ALE) in gastroenterology is mainly based upon its strong antidyspeptic actions which are mediated by its choleretic activity. The aim of this study was to investigate the effects of ALE on bile flow and the formation of bile compounds in anaesthetised Wistar rats after acute and repeated (twice a day for 7 consecutive days) oral administration. A significant increase in bile flow was observed after acute treatment with ALE as well as after repeated administration. The choleretic effects of ALE were similar to those of the reference compound dehydrocholic acid (DHCA). Total bile acids, cholesterol and phospholipid were determined by enzymatic assays. There was a strong ALE-induced increase in total bile acid concentration over the entire experiment. With the highest dose (400 mg/kg), a significant increase was obtained after single and repeated administration. The bile acids-increased effects of ALE were much more pronounced than those of reference (DHCA). No significant differences in cholesterol and phospholipid content could be found. PMID- 12587688 TI - Artichoke leaf extract reduces mild dyspepsia in an open study. AB - A recent post-marketing study indicated that high doses of standardised artichoke leaf extract (ALE) may reduce symptoms of dyspepsia. To substantial these findings, this study investigated the efficacy of a low-dose ALE on amelioration of dyspeptic symptoms and improvement of quality of life. The study was an open, dose-ranging postal study. Healthy patients with self-reported dyspepsia were recruited through the media. The Nepean Dyspepsia Index and the State-Trait Anxiety Inventory were completed at baseline and after 2 months of treatment with ALE, which was randomly allocated to volunteers as 320 or 640 mg daily. Of the 516 participants, 454 completed the study. In both dosage groups, compared with baseline, there was a significant reduction of all dyspeptic symptoms, with an average reduction of 40% in global dyspepsia score. However, there were no differences in the primary outcome measures between the two groups, although relief of state anxiety, a secondary outcome, was greater with the higher dosage (P = 0.03). Health-related quality of life was significantly improved in both groups compared with baseline. We conclude that ALE shows promise to ameliorate upper gastro-intestinal symptoms and improve quality of life in otherwise healthy subjects suffering from dyspepsia. PMID- 12587689 TI - Potentiation by saiboku-to of diazepam-induced decreases in hippocampal and striatal acetylcholine release in rats. AB - Effects of saiboku-to, a traditional oriental herbal medicine, on diazepam induced changes in cerebral acetylcholine (ACh) were investigated in rat striatum and hippocampus. Diazepam (10 mg/kg, i.p.) increased tissue concentrations of the ACh in both regions. The increase was enhanced in rats subacutely treated with saiboku-to (2.0 g/kg, p.o., once a day) for 7 days. Diazepam also decreased release levels of ACh in both regions. The release levels were further decreased in saiboku-to-treated rats. On the other hand, no significant changes in ACh synthesizing and the hydrolyzing enzyme activities in either brain region were observed in saiboku-to-, diazepam- and combination-treated rats. These results suggest that not only is the diazepam-induced increase in tissue ACh due to the inhibition of ACh release but also that saiboku-to potentiates diazepam-induced inhibition of ACh release. PMID- 12587690 TI - Central effects of citral, myrcene and limonene, constituents of essential oil chemotypes from Lippia alba (Mill.) n.e. Brown. AB - Citral, myrcene and limonene (100 and 200 mg/kg body wt., i.p.), constituents of essential oils from Lippia alba chemotypes, decreased not only the number of crossings but also numbers for rearing and grooming, as measured by the open field test in mice. Although muscle relaxation detected by the rota rod test was seen only at the highest doses of citral (200 mg/kg body wt.) and myrcene (100 and 200 mg/kg body wt.), this effect was observed even at the lowest dose of limonene (50 mg/kg body wt.). Also, citral and myrcene (100 and 200 mg/kg body wt.) increased barbiturate sleeping time as compared to control. Limonene was also effective at the highest dose, and although citral did not increase the onset of sleep, it increased the duration of sleep, which is indicative of a potentiation of sleeping time. Citral (100 and 200 mg/kg body wt.) increased 2.3 and 3.5 times, respectively, the barbiturate sleeping time in mice. Similar effects were observed for myrcene and limonene at the highest dose (200 mg/kg body wt.) which increased the sleeping time around 2.6 times. In the elevated plus maze, no effect was detected with citral up to 25 mg/kg body wt., while at a high dose it decreased by 46% the number of entries in the open arms. A smaller but significant effect was detected with limonene (5 mg/kg body wt.). While myrcene (10 mg/kg body wt.) decreased only by 22% the number of entries in the open arms, this parameter was decreased by 48% at the highest dose. Our study showed that citral, limonene and myrcene presented sedative as well as motor relaxant effects. Although only at the highest dose, they also produced a potentiation of the pentobarbital-induced sleeping time in mice, which was more intense in the presence of citral. In addition, neither of them showed an anxiolytic effect, but rather a slight anxiogenic type of effect at the higher doses. PMID- 12587691 TI - Cardiovascular effects of the essential oil of Mentha x villosa in DOCA-salt hypertensive rats. AB - The present study investigated the effects of chronic treatment with deoxycorticosterone-acetate (DOCA)-salt on cardiovascular responses to intravenous (i.v.) injection of the essential oil of Mentha x villosa (EOMV) in conscious rats. In both DOCA-salt-hypertensive and uninephrectomized control, conscious rats, i.v. bolus injections of EOMV (1 to 20 mg/kg body wt.) decreased mean aortic pressure (MAP) and heart rate (HR) in a dose-dependent manner. Treatment with DOCA-salt significantly enhanced EOMV-induced decreases in MAP, without affecting bradycardia. Likewise, both maximal percent and absolute decreases in MAP elicited by i.v. injection of the ganglion blocker, hexamethonium (30 mg/kg body wt.), were significantly greater in DOCA-salt hypertensive than in control rats. In DOCA-salt-hypertensive rats, i.v. pretreatment with hexamethonium (30 mg/kg body wt.) reduced the bradycardia elicited by EOMV (1 to 20 mg/kg body wt.) without affecting the enhancement of EOMV-induced hypotension. These results show that i.v. treatment with EOMV decreases blood pressure in conscious DOCA-salt-hypertensive rats dose dependently, and that this action is enhanced when compared with uninephrectomized controls. This enhancement could be related mainly to an increase in EOMV-induced vascular smooth muscle relaxation, rather than to enhanced sympathetic nervous system activity in this hypertensive model. PMID- 12587692 TI - Anti-inflammatory activity of linalool and linalyl acetate constituents of essential oils. AB - Linalool and linalyl acetate are the principal components of many essential oils known to possess several biological activities, attributable to these monoterpene compounds. In this work, we evaluated individually the anti-inflammatory properties of (-) linalool, that is, the natural occurring enantiomer, and its racemate form, present in various amounts in distilled or extracted essential oils. Because in the linalool-containing essential oils, linalyl acetate, is frequently present, we also examined the anti-inflammatory action of this monoterpene ester. Carrageenin-induced edema in rats was used as a model of inflammation. The experimental data indicate that both the pure enantiomer and its racemate induced, after systemic administration, a reduction of edema. Moreover, the pure enantiomer, at a dose of 25 mg/kg, elicited a delayed and more prolonged effect, while the racemate form induced a significant reduction of the edema only one hour after carrageenin administration. At higher doses, no differences were observed between the (-) enantiomer and the racemate; a further increase in the dose of both forms did not result in an increased effect at any time of observation. The effects of equi-molar doses of linalyl acetate on local edema were less relevant and more delayed than that of the corresponding alcohol. These finding suggest a typical pro-drug behavior of linalyl acetate. The results obtained indicate that linalool and the corresponding acetate play a major role in the anti-inflammatory activity displayed by the essential oils containing them, and provide further evidence suggesting that linalool and linalyl acetate producing species are potentially anti-inflammatory agents. PMID- 12587693 TI - Evaluation of antipyretic activity of leaf extracts of Mallotus peltatus (Geist) Muell. arg. var acuminatus: a folk medicine. AB - A study was carried out to evaluate the anti-pyretic potential of the methanol extract of Mallotus peltatus (Geist) Muell. Arg. var acuminatus leaf, a folk medicine of Onge tribes of Bay Islands, on normal body temperature and yeast induced pyrexia in Wister albino rats. The leaf extract at oral doses of 100, 200 and 300 mg kg(-1), p.o., showed significant reduction in normal body temperature and yeast-provoked elevated temperature in a dose-dependent manner and the anti pyretic effect was comparable to that of standard anti-pyretic agent paracetamol (150 mg kg(-1), p.o.). The effect also extended up to 5 hours after the drug administration. PMID- 12587695 TI - Effect of isorhapontigenin on respiratory burst of rat neutrophils. AB - The effect of Isorhapontigenin (Iso) isolated from Belamcanda chinensis on respiratory burst of rat neutrophils was investigated. Iso (1, 10, 100 mmol/l) showed an inhibitory effect on superoxide anion and hydrogen peroxide production in phorbol myristate acetate (PMA) activated rat neutrophils in a concentration dependent manner. Scanning electron microscopy detected that Iso (100 mmol/l) protected against surface changes in rat neutrophils stimulated with PMA. Also, 100 mmol/l Iso inhibited the release of beta-glucuronidase from the activated neutrophils. Electron-spin resonance (ESR) detected that Iso scavenged oxygen free radicals generated in the PMA activated Neutrophils. These results suggest that Iso inhibits respiratory burst of PMA-activated rat neutrophils by scavenging oxygen free radicals. PMID- 12587694 TI - Evaluation of anti-pyretic potential of Ficus racemosa bark. AB - A study was carried out to evaluate the anti-pyretic effect of a methanol extract of stem bark of Ficus racemosa Linn. (MEFR) on normal body temperature and yeast induced pyrexia in albino rats. A yeast suspension (10 ml/kg body wt.) increased rectal temperature 19 h after subcutaneous injection. The MEFR, at doses of 100, 200 and 300 mg/kg body wt. p.o., showed significant dose-dependent reduction in normal body temperature and yeast-provoked elevated temperature. The effect extended up to 5 h after drug administration. The anti-pyretic effect of MEFR was comparable to that of paracetamol (150 mg/kg body wt., p.o.), a standard anti pyretic agent. PMID- 12587696 TI - Evaluation of anti-diarrheal activity of Cleome viscosa L. extract in rats. AB - A study was undertaken to evaluate the effect of a methanol extract of the entire plant Cleome viscosa L. (CVME) (Family; Capparidaceae) for its anti-diarrheal potential against some of the experimental models of diarrhea in rats. CVME showed significant inhibitory activity against castor-oil-induced diarrhea and PGE2 induced enteropooling in rats. The extract also showed a significant reduction in gastrointestinal motility in the charcoal meal test in rats. The results obtained establish the efficacy and substantiate the folklore claim as an anti-diarrheal agent. PMID- 12587697 TI - Development of a paw paw herbal shampoo for the removal of head lice. AB - The development and clinical testing of an herbal lice removal shampoo, containing a standardized extract of paw paw, thymol, and tea tree oil, are described. All of these ingredients were selected for their ability to deplete adenosine triphosphate (ATP) levels and, thus, prevent ATP-dependent pesticide resistance. Optimum concentrations of the ingredients, treatment times, and dosing schedules were established through in vitro tests with head lice. In addition to pilot studies involving 21 participants, a final clinical trial, using the optimum shampoo formulation in 16 participants, demonstrated 100% effectiveness in removing head lice and nits. PMID- 12587698 TI - Pharmacological activities of crude acetone extract and purified constituents of Salvia moorcraftiana Wall. AB - The crude acetone extract of aerial parts of Salvia moorcraftiana Wall. was screened for various biological activities including Lemna bioassay, antifungal, antibacterial, leishmanicidal, insecticidal activities and brine shrimp cytotoxicity. It was found to possess strong phytotoxic activity against Lemna aequinoctials Welve. and moderate antifungal activity against animal and plant pathogens. The purified chemical constituents were tested for enzyme inhibition activity. Two constituents (compounds 3 and 8) were found to be effective inhibitors of alpha-glucosidase. PMID- 12587699 TI - Tribulosin and beta-sitosterol-D-glucoside, the anthelmintic principles of Tribulus terrestris. AB - Successive extracts of Tribulus terrestris prepared using petroleum ether, chloroform, 50% methanol and water were tested for anthelmintic activity in-vitro using the nematode Caenorhabditis elegans. The activity could be detected only in 50% methanol extract which on further bioactivity guided fractionation and chromatographic separation yielded a spirostanol type saponin, tribulosin and beta-sitosterol-D-glucoside. Both the compounds exhibited anthelmintic activity with ED50 of 76.25 and 82.50 microg/ml respectively. PMID- 12587700 TI - A preliminary RAPD-PCR analysis of Cimicifuga species and other botanicals used for women's health. AB - Traditional taxonomic methods of botanical identification that rely primarily on morphological observations cannot be used efficiently when only powdered plant materials are available. Thus, our objectives were to determine if we could apply a molecular approach to: a) produce unique DNA profiles that are characteristic of the species, and b) determine if the geographical area or time of collection influences these DNA profiles. Towards this end, random amplified polymorphic DNA (RAPD) analyses were performed on a number of botanicals currently used for women's health. The test materials included samples from three species each of the genera Cimicifuga (Actaea) and Trifolium, as well as samples of Vitex agnus castus L., Glycyrrhiza glabra L., Gingko biloba L., Valeriana officinalis L., Angelica sinensis (Oliv.) Diels, Viburnum prunifolium L., Humulus lupulus L., Vaccinium macrocarpon Ait., Panax ginseng C.A. Mey. Cimicifuga racemosa (L.) Nutt. and Trifolium pratense L. are currently under clinical investigation in our basic research laboratories and medical clinic for the relief of post-menopausal symptoms. Characteristic profiles produced with the OPC-15 primer could distinguish the three Cimicifuga species: C. racemosa, C. americana and C. rubifolia. Similar results were obtained with the three Trifolium species: Trifolium pratense L., Trifolium incarnatum L., and Trifolium repens L. Accessions of cultivated T. pratense collected from the same field at different times, produced identical profiles. Accessions of Cimicifuga species collected from different geographical areas produced similar but not identical DNA profiles; however, species-specific DNA fragments were identified. These results demonstrate that RAPD analysis can be applied to distinguish species when only powdered material is available for testing. This methodology can be applied to identify species of commercial value regardless of collection time or geographic area. PMID- 12587701 TI - Case reports--disparaging herbal drugs? PMID- 12587702 TI - The quest for nursing excellence. PMID- 12587703 TI - Differences in cardiovascular disease risk factors in elderly and younger patients with type 2 diabetes in the West Indies. AB - OBJECTIVE: To assess the cardiovascular disease (CVD) risk factors in elderly (> or = 60 years) and younger patients with Type 2 diabetes visiting two primary care clinics in Trinidad. MATERIALS AND METHODS: Fasting blood samples were taken from one hundred and ninety-one (127 females, 64 males) patients with Type 2 diabetes visiting two primary care clinics between 1 January and 30 April 2000. Anthropometric indices, blood pressure, plasma glucose, serum lipids and insulin were measured. Homeostasis Model Assessment (HOMA) was used to assess basal insulin resistance (IR). RESULTS: Of the 191 patients studied, 58.6% were <60 years old while 41.4% were > or = 60 years old. The younger patients had higher prevalence rates of cigarette smoking and use of alcoholic drinks (p < 0.05). The female younger patients had significantly higher mean body mass index (BMI), glycated haemoglobin (HbA1c), triglyceride, total-cholesterol, fasting plasma glucose and IR than the elderly female patients (p < 0.05). Similarly, the younger male patients had significantly higher mean HbA1c, creatinine, fasting plasma glucose, IR and lower HDL-cholesterol levels than the elderly male patients (p < 0.05). Generally, the younger patients had significantly higher prevalence rates of hypertriglyceridaemia, obesity, poorer glycaemia and blood pressure control. CONCLUSION: The results indicate that younger patients withType 2 diabetes had poorer metabolic control and higher prevalence rates of CVD risk factors than the elderly patients. The greater risk of CVD in younger patients was not independent of gender and ethnicity. PMID- 12587704 TI - Unplanned extubation: a local experience. AB - OBJECTIVES: To study the outcome of unplanned extubation (UE) in the Medical Intensive Care Unit (MICU) and to identify factors which predict the need for reintubation. METHODS: A prospective study of all mechanically ventilated patients admitted to MICU in 1998. Patients were enrolled into the study at the point of their first UE. The primary endpoint was reintubation after UE and secondary endpoint was death from any cause during hospitalisation. RESULTS: A total of 543 patients were admitted to MICU of which 312 were mechanically ventilated. UE accounted for 8.7% of our mechanically ventilated patients. The mean APACHE 11 score was 20 (+/- 10), mean time between intubation and UE was 3.1 days (+/- 3.1), mean length of MICU stay was 10.1 days (+/- 10.2) and mean hospital stay was 27.0 days (+/- 36.1). Eighty-seven percent of the UE was deliberate. The rate of reintubation after failed UE was 58.3% of which 71.4% had immediate reintubation. Twenty-nine percent of patients were undergoing weaning during UE. The in-hospital mortality was 25%. All deaths occurred in the group who failed UE. Patients who failed UE had a higher mean APACHE 11 score, a higher mean pre-extubation FiO2 level and a lower mean PaO2/ FiO2 ratio (p < 0.05). CONCLUSION: UE accounted for 8.7% of our mechanically ventilated patients and 58.3% of these patients required reintubation. Failed UE was associated with a higher mortality. A higher APACHE 11 score, higher pre-extubation FiO2 level and a lower PaO2/FiO2 ratio were associated with reintubation after failed UE. PMID- 12587705 TI - Knowledge and practice of breast cancer screening amongst public health nurses in Singapore. AB - AIM OF STUDY: The study aimed to examine the knowledge and practice of breast cancer screening amongst Public Health nurses who are patient educators. METHOD: A self-administered questionnaire was sent to all 447 nurses in the Public Health Service to assess their knowledge and practice of breast cancer screening. Response rate was 96.4%. Knowledge scores ranged from 0-17 with one point given to a correct knowledge question, zero for wrong answer. RESULTS: Median knowledge score was nine and 58.3% of nurses scored > or = 9. Statistically significant factors influencing knowledge scores were related to the nursing profession, namely nursing qualifications, current nursing post and current workplace. 76.2% of Nursing Officers/Higher Nursing Officers and 74.5% of midwives had knowledge scores > or = 9 compared with 57.3% of staff nurses and 40.8% of assistant nurses (p < 0.01). 60.1% of nurses who were taught breast-self examination scored > or = 9 (p < 0.05). As for practice, 93.7% of nurses did breast-self-examination, 54.1% had clinical breast examination in the past one year and 50.2% had mammogram done. On multivariate analysis, significant factors affecting practice of clinical breast examination were marital status (Prevalence Rate Ratio 1.94, 95% Confidence Interval 1.20-3.15), doctor's gender (PRR 1.35, CI 1.04-1.75) and those affecting mammogram were age group (PRR 1.78, CI 1.27-2.48), marital status (PRR 1.63, CI 1.03-2.59), history of breast disease (PRR 1.51, CI 1.06-2.16). CONCLUSIONS: The majority of nurses had certain misconceptions in the knowledge of breast cancer and breast cancer screening. A higher proportion of nurses in the Family Health Service had higher knowledge scores than other nurses in the Public Health Service. Most of the nurses did breast-self-examination. Having ever married and the availability of a female doctor influenced the nurses attending clinical breast examination. Nurses who were > or = 50 years old, ever married and those with a positive history of breast disease were more likely to have mammography done. PMID- 12587706 TI - Risk factors for stroke and predictors of one-month mortality. AB - BACKGROUND: Stroke is the third most common cause of death in Malaysia.The prevalence of risk factors and predictors of mortality of stroke in Malaysia are poorly understood. AIM: To identify the prevalence of major risk factors for stroke and to determine predictors of one-month mortality. METHOD: Prospective study of all stroke patients admitted to Penang Hospital between December 1998 and November 1999. All patients were subjected to brain CT. Predictors of one month mortality: systolic and diastolic hypertension, hyperglycaemia, type of stroke, age > or = 70, poor Glasgow coma score (GCS) on admission and deterioration of score were assessed. RESULTS: A total of 246 (139 male and 107 female) patients were included. Median age was 65 years. Hypertension was the commonest risk factor (71.5%) followed by diabetes mellitus (40.2%) and hyperlipidaemia (37%). 74.8% of the cases were ischaemic in origin and 25.2% haemorrhagic. Mortality at one month was 20.3%. Using multivariate analysis and logistic regression, deterioration of GCS (OR = 46.04), poor GCS on admission (OR = 12.35) and haemorrhagic stroke (OR = 3.45) were independent predictors of one month mortality. CONCLUSION: Hypertension and diabetes mellitus are the commonest risk factors of stroke among patients admitted to a tertiary hospital in Malaysia. Significant predictors of one-month mortality include the admission GCS, deterioration of GCS and haemorrhagic stroke. PMID- 12587707 TI - Unanticipated admission after day surgery. AB - Day surgery is becoming more common due to its cost effectiveness as well as patient acceptance. With increasing caseloads, there is a need to maintain quality of care. The purpose of this study is to identify the reasons for unanticipated admissions in our day surgical population, with the aim of improving efficiency of day surgical services, yet maintaining a high standard of patient care. A retrospective review of records of patients who were admitted over the two-year study period was conducted. Unanticipated admission was defined as unplanned admission after a day surgical procedure. Data relating to physical status, perioperative complications and reasons for hospital admission were recorded. A total of 10,801 procedures were done, and 163 patients were admitted. The unanticipated admission rate was 1.5%. Most of the admissions were surgically related (62.8%), followed by anaesthesia (12.2%), social (9.5%) and medical reasons (8.1%). Seventy-five percent of these admissions were potentially preventable. The majority were due to common problems like postoperative pain, admission for surgical observation and for social reasons. Non preventable causes (25%) were mainly due to unrelated medical problems. PMID- 12587708 TI - A case of generalised cutaneous granulocytic sarcoma in an elderly patient with myelodysplastic syndrome. AB - Granulocytic sarcoma is a rare extramedullary malignant mass composed of primitive cells of the granulocytic lineage. It can arise from any part of the body and is frequently associated with haematological diseases, commonly acute myeloid leukaemia. Rarely, it has been found in conjunction with myelodysplastic syndrome. We report a case of cutaneous granulocytic sarcoma in a 73-year-old lady. The patient presented with a two-month history of multiple skin nodules which were confirmed by skin biopsy to be granulocytic sarcoma. Bone marrow examination was consistent with myelodysplastic syndrome. Localised radiotherapy to the skin lesions were given. She died from septicaemia six months after presentation. The management of this condition presents a diagnostic and therapeutic dilemma for both the pathologist and physician. In cases which are poorly differentiated as in this case, histological diagnosis is particularly difficult. Its definitive diagnosis would then require the additional use of a broad panel of immunohistochemical and cytochemical stains. PMID- 12587709 TI - Risperidone and megacolon. AB - Risperidone is an atypical anti-psychotic medication with both 5HT2 receptor and D2 dopamine receptor antagonism. Its use has been reported to be generally safe with very few gastro-intestinal (GI) adverse or side effects. In this paper, we describe a case of megacolon associated with the use of risperidone. A 44-year old man suffering from schizophrenia was treated with risperidone and developed gross abdominal distension after twenty-five days. Abdominal X-ray and colonoscopy showed megacolon. He improved following a surgical decompression and a reduction of risperidone dosage. We discuss the neuro-electro-physiological mechanisms of gastro-intestinal motility and conclude that the risperidone associated megacolon may be dose-related and that there should be a heightened awareness of such possible GI complication when using risperidone. PMID- 12587710 TI - Diagnosis disclosure in cancer patients--when the family says "no!". AB - It is not uncommon for family members to thwart a doctor's attempt at disclosure of cancer diagnosis to the patient. This stems from concern and love as well as fear of the negative psychological impact of such revelation. Indeed, collusion of this nature was the norm in medical practice till the not too distant past. In this era of patient autonomy and the right to information, we must be careful not to allow attitudes of medical staff or patients' families, language difficulties and organisational issues become barriers to patient's access to information and their psychological coping of the illness. At the same time, we also need to be acutely aware of the unique cultural environment we work in and the families' emotional needs even as we aim to match the patient's desire for information. PMID- 12587711 TI - Article: Somatic cell nucleartransfer (cloning): implications for the medical practitioner. PMID- 12587712 TI - Clinics in diagnostic imaging (78). Cardiac rhabdomyoma in tuberous sclerosis. AB - A seven-day-old boy with a family history of tuberous sclerosis presented with congestive heart failure. Cardiac-gated magnetic resonance (MR) imaging showed a mass in the right atrium, which was T1-isointense and T2-hyperintense relative to myocardium. Follow-up echocardiographical studies showed a decrease in the number and size of the nodules. Finally, no cardiac nodules were seen when he was two years and eleven months old. The echocardiographical studies, MR imaging findings and the clinical history are consistent with cardiac rhabdomyomas. MR imaging of the brain also showed multiple subependymal nodules, white matter abnormalities, and cortical tubers. The varied imaging appearances of tuberous sclerosis are discussed and illustrated with additional examples. PMID- 12587713 TI - Immune system and antioxidants, especially those derived from Indian medicinal plants. AB - During the functioning of the immune system, such as in phagocytosis, reactive oxygen and nitrogen species are generated. If they are left unchecked they can affect the components of the immune system by inducing oxidative damage. This is more so in the elderly or during inflammation where there is excess generation of these reactive species than can be taken care of by the defenses in the form of antioxidants. Dietary supplementation with antioxidants may greatly help in such conditions. There are some indications of possible benefits of antioxidant supplementation. Natural compounds from medicinal plants having antioxidant and immunomodulatory activities have potential as therapeutic agents in this regard. Indian medicinal plants with these activities have been identified and their antioxidant and immunomodulatory effects reviewed. The possible future prospects in this regard are also outlined. PMID- 12587714 TI - Oxidative stress and experimental carcinogenesis. AB - The focus of this review is to provide state-of-the-art knowledge on the involvement of oxygen free radicals (OFR) in carcinogenesis with a particular reference to skin model system as the process of cancer development is best understood in this organ. However, a brief description of the role of OFR in other organs is also provided. The term OFR refers to forms of oxygen exhibiting high reactivity and having at least one unpaired electron. The role of OFR in different stages of carcinogenesis such as initiation, promotion and progression is described. Out of many mechanisms described for the chemical initiation of tumorigenesis, a number of them may involve free radicals in the cascade of reactions. Evidences that support the involvement of free radicals in tumor promotion include (i) a number of free radical-generating compounds are found to be tumor promoters in various animal model systems, (ii) ROS generating systems can mimic the biochemical action of tumor promoters, (iii) some tumor promoters stimulate the production of ROS, (iv) tumor promoters modulate the cellular antioxidant defense systems, and (v) free radical scavengers, detoxifiers and antioxidants inhibit the process of tumor promotion. The role of ROS in the progression stage of carcinogenesis is evident from the fact that a number of different free radical generating compounds enhance the malignant conversion of benign papillomas into carcinoma and their effectiveness may be related to the type of radicals produced into the biological system. PMID- 12587715 TI - Melatonin oxidative stress and neurodegenerative diseases. AB - Oxidative Stress is implicated as one of the primary factors that contribute to the development of neurodegenerative diseases like Alzheimer's Disease, Parkinsonism and neurological conditions like epileptic seizures, stroke, brain damage, neurotrauma etc. The increased formation and release of oxygen free radicals coupled with the rather low antioxidative potential of the central nervous system are the major reasons that account for the enhanced oxidative stress seen in neuronal cells. In addition to this, brain is also enriched with polyunsaturated fatty acids that render neuronal cells easily vulnerable to oxidative attack. The fact that there is increased incidence of neurodegenerative disorders in aged individuals, has prompted many investigators to search for a common factor whose progressive decline with increase in age could account for increased oxidative stress resulting in senescence and age associated degenerative diseases. Since melatonin, the hormone secreted from the pineal gland has a remarkable anti-oxidant property and whose rate of production declines with increase in age, has prompted many to suggest that this hormone plays a crucial role in the genesis of neurodegenerative diseases. Melatonin cannot only scavenges oxygen free radicals like super oxide radical (O2-), hydroxyl radical (*OH), peroxyl radical (LOO*) and peroxynitrite anion (ONOO-), but can also enhance the antioxidative potential of the cell by stimulating the synthesis of antioxidative enzymes like super oxide dismutase (SOD), glutathione peroxidase (GPX), and also the enzymes that are involved in the synthesis of glutathione. In many instances, melatonin increases the expression of m RNA's of the antioxidative enzymes. Melatonin administration has been shown to be effective in counteracting the neurodegenerative conditions both in experimental models of neurodegenerative diseases and in patients suffering from such diseases. A disturbance of melatonin rhythm and secretion also has been noted in patients suffering from certain neurodegenerative diseases. From all these, it is evident that melatonin has a neuroprotective role. PMID- 12587716 TI - Biological significance of singlet oxygen. AB - The biological significance of singlet oxygen (1O2), an electronically excited species of oxygen, has been realized only in the last two decades. This was mainly due to the lack of proper methodology to generate this reactive oxygen species (ROS) in pure form and its reactions with biological molecules. Recent studies, using newly developed detection methods, show that 1O2 being generated in many biological systems, can significantly and quite often adversely alter several crucial biomolecules including DNA, proteins and lipids with undesirable consequences including cytotoxicity and/or disesase development. The reactions of 1O2 with the biological molecules are rather specific, as compared to other ROS. There are various compounds, mainly derived from natural sources that offer protection against damage induced by 1O2. Among the antioxidants carotenoids are the most effective singlet oxygen quenchers followed by tocopherols and others. The same reactive species if generated specifically in diseased states such as cancer can lead to the cure of the disease, and this principle is utilized in the newly developing modality of cancer treatment namely photodynamic therapy. Singlet oxygen, in low concentrations can also act as signaling molecule with several biological implications. This review clearly brings out the biological significance of 1O2. PMID- 12587717 TI - Involvement of reactive oxygen species in gastric ulceration: protection by melatonin. AB - Uncontrolled hydrochloric acid secretion and ulceration in the stomach due to various factors are serious global problems today. Although the mechanism of acid secretion from the parietal cell is now fairly known, the mechanism of gastric ulceration is still not clear today. Among various causes of gastric ulceration, lesions caused by stress, alcohol consumption, Helicobacter pylori infection and use of nonsteroidal antiinflammatory drugs have been shown to be mediated largely through the generation of reactive oxygen species especially hydroxyl radical (*OH). A number of excellent drugs have been proved useful in controlling hyperacidity and ulceration but their long term uses are not devoid of disturbing side-effects. Hence, the search is still on to find out a compound possessing antisecretory, antiulcer and antioxidant properties which will serve as a powerful therapeutic agent to cure gastric hyperacidity and ulcer. This article describes the role of reactive oxygen species in gastric ulceration, drugs controlling them with their merits and demerits and, the role of melatonin, a pineal hormone in protecting the gastric lesions with a final commentary on how melatonin research with respect to gastric pathophysiology can be taken forward with a view to projecting this indole as a promising therapeutic agent to control gastric ulceration in humans. PMID- 12587718 TI - Inevitable glutathione, then and now. AB - Glutathione a predominant tripeptide thiol compound of many prokaryotes and eukaryotes, is synthesized from its precursor amino acids eg. gamma-glutamate, cysteine and glycine. It is mainly involved in detoxication mechanisms through conjugation reactions. Other functions include thiol transfer, destruction of free radicals and metabolism of various exogenous and endogenous compounds. It becomes mandatory for a cell to manage high concentration of intracellular GSH to protect itself from chemical/dug abuse. Glutathione dependent enzymes viz: glutathione-S-transferases, glutathione peroxidase, glutathione reductase and gamma-glutamate transpeptidase facilitate protective manifestations. Liver serves as a glutathione-generating factor which supplies the kidney and intestine with other constituents of glutathione resynthesis. The principal mechanism of hepatocyte glutathione turnover appears to be cellular efflux. Kidney too plays an important role in organismic GSH homeostasis. Role of GSH in organs like lung, intestine and brain has recently been described. GSH involvement in programmed cell death has also been indicated. Immense interest makes the then "thee glutathione" as "inevitable glutathione". This article describes the role of this vital molecule in cell physiology and detoxication mechanisms in particular. PMID- 12587719 TI - Mechanistic pathways of antioxidant cytoprotection by a novel IH636 grape seed proanthocyanidin extract. AB - To understand the bioavailability and mechanistic pathways of cytoprotection by IH636 grape seed proanthocyanidin extract (GSPE, commercially known as ActiVin) a series of in vitro and in vivo studies were conducted. Comparative protective abilities of GSPE, and vitamins C and E, singly and in combination, were assessed against smokeless tobacco extract (STE)-induced oxidative stress, DNA fragmentation and apoptotic cell death in a primary culture of normal human oral keratinocytes. GSPE protected against STE-induced oxidative stress, DNA damage and apoptotic cell death, and provided better protection as compared to vitamins C and E, singly and in combination. The bioavailability and protective ability of GSPE were examined against acetaminophen (AP)-induced hepato- and nephrotoxicity, amiodarone (AM)-induced lung toxicity, doxorubicin (DX)-induced cardiotoxicity and dimethylnitrosamine (DM)-induced spleenotoxicity in mice. GSPE-fed animals were compared with GSPE-untreated mice to evaluate the protective ability of GSPE against these structurally diverse drugs/chemicals. Serum chemistry changes, histopathology and DNA damage were evaluated. Results indicate that GSPE preexposure prior to the drugs/chemicals such as AP, AM, DX or DM treatment, provided near complete protection in terms of serum chemistry changes and inhibition of both forms of cell death, e.g., apoptosis and necrosis. DNA damage in various tissues triggered by these agents was significantly reduced in GSPE fed animals. Histopathological examination of multiple target organs provided similar data. The results suggest that GSPE exposure is bioavailable and provides significant multiorgan protection against structurally diverse drug- and chemical induced toxic assaults. Further, these studies exhibited a series of mechanistic information including free radical scavenging ability, anti-endonucleolytic activity, cytochrome P450 2E1 inhibitory activity, anti-necrotic, anti-apoptotic and anti-carcinogenic activities, modulatory effects on antioxidative and apoptotic regulatory genes such as Bcl2, c-myc and p53, which may be responsible for the novel chemoprotective properties exhibited by GSPE. PMID- 12587720 TI - Free radical scavenging and metal chelation by Tinospora cordifolia, a possible role in radioprotection. AB - Aqueous extract of T. cordifolia inhibited Fenton (FeSO4) reaction and radiation mediated 2-deoxyribose degradation in a dose dependent fashion with an IC50 value of 700 microg/ml for both Fenton and radiation mediated 2-DR degradation. Similarly, it showed a moderate but dose dependent inhibition of chemically generated superoxide anion at 500 microg/ml concentration and above with an IC50 value of 2000 microg/ml. Aqueous extract inhibited the formation of Fe2+ bipiridyl complex and formation of comet tail by chelating Fe2+ ions in a dose dependent manner with an IC50 value of 150 microg/ml for Fe2+-bipirydyl formation and maximally 200 microg/ml for comet tail formation, respectively. The extract inhibited ferrous sulphate mediated lipid peroxidation in a dose-dependent manner with an IC50 value of 1300 microg/ml and maximally (70%) at 2000 microg/ml. The results reveal that the direct and indirect antioxidant actions of T. cordifolia probably act in corroboration to manifest the overall radioprotective effects. PMID- 12587721 TI - Effect of antioxidants (vitamin C, E and turmeric extract) on methimazole induced hypothyroidism in rats. AB - The study was to investigate the protective effect of antioxidants against methimazole (MMI) induced hypothyroidism in rats. Male Wistar rats were fed MMI, MMI plus vitamin C, MMI plus vitamin E and MMI plus turmeric extract (TE) supplemented diet. At the end of the experiments, thyroid weights, thyroxine (T4), triiodothyronine (T3) and cholesterol levels were determined. It was observed that MMI treated rats showed increase in thyroid weights, very low levels of circulating T4, T3 and increased levels of total cholesterol as compared to controls (P< 0.001). However, rats which received Vit. C, Vit. E or TE along with MMI showed reduced weights (38-55% less) in thyroid glands (P < 0.01), less suppressed T4 and T3 levels (2-6% and 7-35% respectively) and less increase in total cholesterol levels (19-52%) which are statistically significant. The data suggest the positive effect of antioxidants on thyroid gland which could be due to direct involvement of antioxidants on thyroid gland. PMID- 12587722 TI - Caffeine, quercetin and alizarin stimulate the exhalation of metabolic products of [14C]-N-nitrosodiethylamine in mice. AB - Naturally occurring plant products belonging to different chemical classes namely alizarin, an anthraquinone, caffeine, a methylxanthine derivative and quercetin, a flavonol were studied for their effect on elimination of metabolites of [14C]-N nitrosodiethylamine (14C-NDEA) through respiration in mice. Treatment with caffeine, quercetin and alizarin at doses of 200, 9 and 9 microg/ml respectively, in drinking water enhanced the exhalation of 14CO2, one of the major end products of NDEA metabolism. Radioactive CO2 exhaled in 60 min increased by 2, 1.61 and 1.4-folds in animals treated with caffeine, quercetin and alizarin for 8 weeks respectively. This increase in exhalation in caffeine-treated animals was achieved even in 2 weeks. These compounds had no adverse effects on the absorption of radioactive NDEA from the gut of the animals as shape and time of 14CO2 peak was similar in i.p. and orally administered [14C-NDEA]. Increased detoxification/elimination of the carcinogen could be one of the mechanisms for the anticarcinogenic properties of these phytochemicals in lung tumorigenesis induced by orally administered NDEA. PMID- 12587723 TI - National seminar on biotechnology--challenges and prospects. PMID- 12587724 TI - Nitrogen control of bacterial signal production in Rhizobium meliloti-alfalfa symbiosis. AB - Under nitrogen-depleted conditions nitrogen-fixing soil bacteria of the family Rhizobiaceae are able to induce symbiotic nodules on the roots of leguminous plants where bacteroids convert atmospheric nitrogen to ammonia. The presence of exogenous nitrogen source inhibits the development and the functioning of bacterium-plant symbiosis. Earlier experiments demonstrated that nitrate inhibited all stages of symbiotic interaction, affecting primarily the host functions. The investigation of the possible involvement of the microsymbiont in nitrogen regulation showed that two signalling steps were controlled by ammonium. The synthesis of the first bacterial signal, the Nod factor was repressed by ammonium. The nitrogen signal is conveyed to nodulation (nod) genes by the general nitrogen regulatory (ntr) system and by the nodD3-syrM self-amplifying system. The fine control also involves a negative regulatory factor, ntrR. When ntrR is mutated, more efficient nodule formation and nitrogen fixation is observed in symbiosis with alfalfa even in the presence of ammonium. The biosynthesis of the second bacterial signal succinoglycan is also controlled by ammonium. SyrM, a common regulatory factor for nod and exo gene expression, may contribute to the adjustment of the amount of succinoglycan and the ratio of its biologically active form. PMID- 12587725 TI - DNA mismatch repair, microsatellite instability and cancer. AB - Mismatch (MMR) repair system plays a significant role in restoration of stability in the genome. Mutations in mismatch repair genes hamper their activity thus bring about a defect in mismatch repair (MMR) mechanism thereby conferring instability in the microsatellite sequences of both the coding and non-coding regions of the genome. Mutated mismatch repair genes result in the expansion or contraction of microsatellite sequence and confer microsatellite unstable or replication error positive phenotype. Hypermethylation of promoter regions of some of the MMR genes also causes inactivation of these genes and thus contribute to MSI. Microsatellite instability is an indicator of MMR deficiency and is a prime cause of varied tumorogenesis. PMID- 12587726 TI - Role of macrophage-colony stimulating factor and osteoclast differentiation factor in osteoclastogenesis of bone marrow derived stem cells. AB - Macrophage colony stimulating factor (M-CSF) and osteoclast differentiation factor (ODF) regulate osteoclastogenesis in vivo. Regulation of osteoclast development in vitro by these cytokines has been reported in the present study. Simultaneous addition of ODF and M-CSF during initiation of bone marrow culture inhibited osteoclastogenesis. However, delayed addition of ODF (three days after initiation of the culture) resulted in dramatic increase in phenotypically and functionally mature osteoclast cells. Delayed addition of ODF beyond day three decreased osteoclastogenesis. Further, removal of M-CSF as early as day three inhibited ODF-induced osteoclastogenesis. These studies provided evidence for the importance of co-ordinated regulation of osteoclastogenesis by M-CSF and ODF. PMID- 12587727 TI - Nuclear zinc in liver of cadmium and zinc treated rats. AB - X-ray microanalysis was performed to detect quantitatively, the variation of the nuclear zinc in the liver cells of rats. The nuclear zinc concentration showed statistical decrease and increase in response to cadmium and zinc treatments, respectively. The results suggest that the liver responds differently to cadmium and zinc treatments. The difference in response to either treatment may reflect different mechanisms of zinc transport and metabolism in the liver. The difference in binding affinity of metallothionein (MT) may suggest the involvement of Mt in the metabolism and transport of zinc, an effect, which may be modified by treatment. PMID- 12587728 TI - Antidiabetic and antioxidant effects of S-methyl cysteine sulfoxide isolated from onions (Allium cepa Linn) as compared to standard drugs in alloxan diabetic rats. AB - Antidiabetic and antoxidant effects of S-methyl cysteine sulfoxide (SMCS) isolated from A. cepa and two standard drugs, glibenclamide and insulin were studied and compared in alloxan diabetic rats after using each of them for treatment for two months. These drugs ameliorated the diabetic condition significantly, viz. maintenance of body weight and control of blood sugar in rats. Further they lowered the levels of malondialdehyde, hydroperoxide and conjugated dienes in tissues exhibiting antioxidant effect on lipid peroxidation in experimental diabetes. This is achieved by their stimulating effects on glucose utilization and the antioxidant enzymes, viz. superoxide dismutase and catalase. The probable mechanism of action of SMCS and glibenclamide may be partly dependent on the stimulation of insulin secretions and partly due to their individual actions. In the amelioration of diabetes the standard drugs showed a better action, but as an antioxidant SMCS proved to be a better one. PMID- 12587729 TI - Effect of oxidative stress on serum and antioxidant enzymes in liver and kidney of rats and their modulation through dietary factors. AB - Modulatory effect of a formulated diet based on cereals, pulses and spices incorporated with crude palm oil (CPO), soybean oil (SBO) or cod liver oil (CLO) at 10% dietary level on oxidative stress and antioxidant enzymes was studied in liver and kidney tissues. Activity of alkaline phosphatase (ALP) and acid phosphatase (ACP) increased significantly in serum in various experimental groups. Significant increase in hepatic antioxidant enzymes, catalase, glutathione peroxidase (GPx) was also seen in the experimental groups. SOD activity showed a mixed response. Further, kidney antioxidant enzymes did not show much change compared to those in liver. The results indicated dietary lipid as the key players in determining cellular susceptibility to oxidative stress, which could be modulated by cereals, pulses and spices in the diet. PMID- 12587730 TI - Response of liver antioxidant system to taurine in rats fed high fructose diet. AB - Fructose-fed rats were more susceptible to peroxidative damage as measured by thiobarbituric acid reactive species. The concentrations of lipid peroxides, diene conjugates, lipofuscin and hydroperoxides were significantly higher. The levels of enzymic antioxidants such as vitamin C, vitamin E and glutathione and activities of antioxidant enzymes were significantly lower in fructose-fed rats. When these rats received taurine in drinking water, peroxidative damage was minimal in both plasma and liver. Taurine was effective in inducing the antioxidant potential in fructose-fed rats. Increased peroxidative damage in liver is likely to be associated with fructose dependent pathology, which could be reduced by taurine by enhancing the antioxidant potential. PMID- 12587731 TI - Efficiency of Coleus aromaticus extract in modifying cyclophosphamide and mitomycin-C induced clastogenicity in mouse bone marrow cells. AB - The anticlastogenic potency of the ethanolic extract of a medicinal plant, C. aromaticus was investigated by taking bone marrow chromosomal aberration assay and micronucleus (MN) test as the test parameters. Swiss albino mice were fed orally with different doses (10,15, 25, 50 and 100 mg/kg body weight) of ethanolic extract for 7 days and on the 7th day, two doses each of anticancer drugs cyclophosphamide (CP; 25 and 50 mg/kg body weight) and mitomycin-C (MMC; 4 and 8 mg/kg body weight) were injected, ip, to different groups of animals. Bone marrow MN preparations were made at 24 and 48 hr time intervals. Coleus extract reduced CP and MMC induced MN and lower doses of the extract were found to be more effective than higher doses. The effective doses of extract in MN test were selected to study the anticlastogenic effects against CP (25 and 50 mg/kg body weight) and MMC (2 and 4 mg/kg body weight) induced chromosomal aberrations. The results indicate the protective effect of C. aromaticus against CP and MMC induced cytogenetic damage. PMID- 12587732 TI - Comparative immunopotentiating properties of saponin and incomplete Freund's adjuvant coupled to affinity purified larval antigen of Hyalomma anatolicum anatolicum. AB - Comparative immunostimulatory properties of saponin and incomplete Freund's adjuvant (IFA) were studied using affinity purified 39 kDa larval antigen of Hyalomma anatolicum anatolicum. Significant antibody response to 39 kDa antigen was detected in the sera of rabbits immunized with both 39 kDa plus saponin and 39 kDa plus IFA in comparison to their corresponding control animals. Insignificant differences were noted in the antibody response between the animals of two immunized groups. Upon challenge, the animals immunized with 39 kDa plus IFA rejected 76.2 +/- 9.7% of larvae and 45.8 +/- 4.1% of adults while in group of animals injected with 39 kDa plus saponin rejected 80.9 +/- 11.2% of larvae and 47.2 +/- 5.7% of adults. PMID- 12587733 TI - Beta-fructofuranosidase production by 2-deoxyglucose resistant mutants of aspergillus niger in submerged and solid-state fermentation. AB - Aspergillus niger produces extracellular beta-fructofuranosidase under submerged (SmF) and solid state fermentation (SSF) conditions. After UV mutagenesis of conidiospores of A. niger, 2-deoxyglucose (10 g/l) resistant mutants were isolated on Czapek's minimal medium containing glycerol as a carbon source and the mutants were examined for improved production of beta-fructofuranosidase in SmF and SSF conditions. One of such mutant DGRA-1 overproduced beta fructofuranosidase in both SmF and SSF conditions. In SmF, the mutant DGRA-1 showed higher beta-fructofuranosidase productivity (110.8 U/l/hr) than the wild type (48.3 U/l/hr). While in SSF the same strain produced 322 U/l/hr of beta fructofuranosidase, 2 times higher than that of wild type (154.2 U/l/hr). In SmF, both wild type and mutants produced relatively low level of beta fructofuranosidase in medium containing sucrose with glucose than from the sucrose medium. However in SSF, the DGRA-1 mutant grown in sucrose and sucrose+ glucose did not show any difference with respect to beta-fructofuranosidase production. These results indicate that the catabolite repression of beta fructofuranosidase synthesis is observed in SmF whereas in SSF such regulation was not prominent. PMID- 12587734 TI - Infra-red spectroscopic analyses of banana waste degraded by oyster mushroom. AB - Carbon, hydrogen and nitrogen analyses of banana leaf and pseudostem biomass revealed their potentiality as substrates for microorganisms. Infra-red (IR) spectra of both biomass show presence of cellulose, xylan and lignin. IR spectra of leaf and pseudostem biomass degraded in solid state fermentation (SSF) by two Pleurotus species (P. sajor-caju and P. ostreatus) for 40 days showed the utilization of cellulose, xylan and lignin by these microbes. Dynamics of various lignocellulolytic enzymes of Pleurotus species and analyses of carbon, hydrogen and nitrogen contents of degraded biomass supported the same. Both the Pleurotus species exhibited lignin consumption ability on both the substrates. PMID- 12587735 TI - A novel strategy of oxygen restriction by some diazotrophic enteric bacteria. AB - Protection of nitrogenase against oxygen inactivation in diazotrophs involves numerous strategies. Glutathione is known to play an important role in scavenging oxyradicals in many living systems. The involvement of glutathione (reduced) (GSH), glutathione peroxidase (GPX) and glutathione reductase (GR) in the protection of nitrogenase in free living diazotrophs is reported here for the first time. Reduced glutathione content and the activity of glutathione peroxidase and glutathione reductase increased with increase in oxygen concentration under nitrogen fixing conditions but decreased under anaerobic and nitrogenase repressed conditions. This correlation is used to postulate a protecting role for GSH-GPX-GR system against oxygen inactivation of nitrogenase. PMID- 12587736 TI - In vitro plantlet regeneration from seedling nodal explants of Acacia catechu. AB - Multiple shoots were initiated after 20 days in stem nodes excised from in vitro grown seedlings of Acacia catechu, on Murashige and Skoog's medium adjuvanted with 1 to 100 microM of N6-benzyladenine (BA). Explants were subcultured on the same medium augmented with 1.5 g l(-1) of polyvinylpyrrolidone (PVP) after 30 days. In the second subculture, after 30 days, the explants were transferred to a medium lacking PVP, but containing 10 microM of BA, where nine or ten shoots differentiated per explant within next 30 days. If individual shoots along with some callus were subcultured on BA (10 microM), nearly 15 shoots per explant regenerated in 90 days. Thus, the average number of shoots obtained from each node was 142 after 180 days. Since a seedling develops four nodes after 20 days, theoretically an average of 568 shoots can be obtained from a single seed. If shoots were individually subcultured on 1/2-strength MS medium with 14.7 microM of indole-3-butyric acid (IBA), roots developed in 20 days. Addition of 40 mg l( 1) of glutamic acid to the rooting medium prevented leaf senescence. These plantlets thrived well in garden soil, sand and silica (1:1:1). PMID- 12587737 TI - Esterase as molecular marker for salt tolerance in regenerated plants of rice, Oryza sativa L. AB - Esterase variation was studied in plants regenerated from callus cultures of four rice (Oryza sativa) varieties, viz. pokkali, which is a moderately salt tolerant variety and three salt sensitive varieties MI 48, annapoorna and jyothi. Variation was studied at tillering stage of plants regenerated from callus culture and germinated from seeds. Somaclonal variants for salt tolerance could be detected using variation in esterase banding pattern and activity. PMID- 12587738 TI - Bioregulation of carbohydrate metabolism in relation to source-sink operation during grain-filling phase of growth in wheat. AB - Mobilization of free sugars from vegetative tissues to grain and their transformation to starch in relation to activities of some relevant enzymes during growth and development were investigated in wheat (Triticum aestivum L.). Vegetative tissues, viz. flag-leaf, flag-leaf sheath, nodes and internodes contained high concentration of free sugars from 70 DAS to 18 DPA and that was in the order of accumulation--flag-leaf sheath> flag-leaf and internodes > nodes. In these tissues, major portion of 14C appeared in endogenous sucrose, irrespective of the nature of (U-14C]-sugars supplied. In photosynthetic structures above flag leaf node, namely peduncle, rachis and bracts, the free sugar make-up was maximum at anthesis (90 DAS). Activity of soluble acid invertase (EC 3.2.1.26) was high in these tissues during early stages of grain growth but reverse was true for soluble neutral invertase (EC 3.2.1.27) activity. In apical and basal portions of grain, free sugars were more or less similarly distributed in concentration. Linear and rapid accumulation of starch in endosperm paralleled with a decline in accumulation of this polymer in pericarp-aleurone. In the latter tissue, the activities of starch hydrolyzing enzymes, i.e alpha- and beta-amylases (3.2.1.1 and 3.2.1.2) were high during initial stages of grain growth. During active grain filling, alkaline inorganic pyrophosphatase (EC 3.6.1.1) seemed to play a vital role during starch accumulation in endosperm, whereas the involvement of 3-PGA phosphatase (EC 3.1.3.38) was almost confined to pericarp-aleurone. Impairement of ear head photosynthesis by shading depressed starch synthesis (approximately 50%) indicating, thereby, the significant role of current photosynthates during grain-filling. The results suggested that grain growth in wheat was influenced by an efficient operation of source as well as regulatory factors, including enzymes, constituting intrinsic potential of grain sink. PMID- 12587739 TI - In vitro formation of organ specific proximate carcinogen of benzo(a)pyrene by rat homogenates. AB - In order to determine the organ specific carcinogenicity of benzo(a)pyrene (B(a)P), its metabolites, formed in vitro by incubation with the homogenates from liver, lungs, kidneys, intestine and brain of rats, were isolated by TLC and spectroscopy. B(a)P was found to be converted into a number of metabolites by different tissue homogenates. The results showed that the proximate carcinogenic metabolite, 7,8-dihydro-7,8-dihydroxy B(a)P was formed only when rat lung and kidney homogenates were incubated with B(a)P in vitro. The UV spectral analysis also confirmed the formation of this metabolite only on incubation of B(a)P with rat lung and kidney homogenates. As the proximate carcinogenic metabolite was only formed by incubating B(a)P with the homogenates from target organs, its organ specific carcinogenicity may be explained. PMID- 12587740 TI - Impact of hypercholesterolemia on in vitro toxicity of N-nitrosodiethylamine: effect on lipidperoxidation of blood and tissue. AB - In vitro treatment of erythrocytes of normal and hypercholesterolemic rats with N nitrosodiethylamine (NDEA), an important carcinogen frequently present in human environment and food chain resulted in a marginal increase in osmotic fragility of erythrocytes without affecting their antioxygenic potential as evidenced by insignificant effect on lipid peroxidation (LPO). However, (LPO) of all the tissues (heart, lung, liver, kidney and spleen) increased significantly on in vitro treatment with NDEA. The effects were different in different tissues under different dietary conditions PMID- 12587741 TI - Frequency of feeding and formation of bone growth marks in frog, Rana cyanophlyctis (Schn.). AB - Frogs, R. cyanophlyctis (n = 45) divided into four groups, were exposed to different feeding regimens (live guppies were used as food) such as, daily, alternate day, every fourth day and weekly feeding for 5 months, during wet months of the year (April-September). Two toe clippings were made, one at the beginning and the other at the termination of the experiment. Clipped toes were demineralized, and processed for histology. In 6 out of 45 frogs one line of arrested growth (LAG) was present in the phalangeal histology at the beginning of the experiment while, at the termination of experiment 34 out of 43 frogs exhibited one LAG each indicating that in 26 frogs LAG appeared freshly during the experimental period. The fact that LAGs are formed in regularly fed frogs suggests the humid weather/seasonal rainfall may play relatively important role than the feeding in cyclic bone growth and formation of growth marks in this frog. PMID- 12587742 TI - Urinary proteins and pheromonal communication in mammals. AB - Urinary proteins play a significant role as pheromones and pheromone-binders in mammalian reproduction and social behaviour. The present study was carried out to quantify the urinary proteins in five different mammalian species viz mouse, rat, rabbit, bovine and human. The results revealed that the male rodents excrete large amounts of urinary protein as compared to that of other mammals. In addition, the male mammals excrete a higher quantity of protein than do the females., suggesting the role of androgens in excretion of protein. The presence of higher concentration of urinary proteins in rodents suggests that the rodents depend more on urinary proteins for olfactory/social communication. PMID- 12587744 TI - Production of glycolipids containing biosurfactant by Pseudomonas species. AB - Microorganisms, that degrade hydrocarbon were isolated and screened for their biosurfactant activity. A total of 68 strains were isolated and tested for their glycolipid activity of which 4 isolates showed good glycolipid activity. Isolate K10 gave the maximum biosurfactant production in medium A (containing kerosene as a sole carbon source) as compared to medium B (containing glucose as a sole carbon source). Characterization of isolate K10 showed that it belongs to Pseudomonas species. PMID- 12587743 TI - Ocimum sanctum aqueous leaf extract provides protection against mercury induced toxicity in Swiss albino mice. AB - HgCl2 (5.0 mg/kg body weight) induced toxicity led to significant elevation of lipid peroxidation (LPO) level but decline in the glutathione content in liver of Swiss albino mice. In serum of HgCl2 treated mice there was significant elevation in serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) activities but significant decline in the alkaline phosphatase activity. Animals treated with O. sanctum extract (10 mg/kg body weight, po) before and after mercury intoxication showed a significant decrease in LPO level, SGOT and SGPT activities and increase in serum alkaline phosphatase activity and glutathione (GSH) content. Ocimum treatment alone did not alter SGOT, SGPT and alkaline phosphatase activities but significantly enhanced reduced glutathione. The results suggest that oral administration of Ocimum extract provides protection against HgCl2 induced toxicity in Swiss albino mice. PMID- 12587745 TI - Monoxenic in vitro production and colonization potential of AM fungus Glomus intraradices. AB - The paper reports the establishment of mycorrhizal infection of a non-mycorrhizal Ri-T-DNA transformed carrot root when co-cultured with a surface sterilized sweet potato root segment colonized by arbuscular mycorrhizal (AM) fungus G. intraradices on minimal M medium. Extensive fungal hyphal emergence from each cut end of the mycorrhizal sweet potato root piece was observed in one week old cultures. These hyphae caused infection on contacting the transformed-carrot- root segment and produced many hyphae and spores both inside and outside the zone of the root after 6 week of growth. Axenically produced fungal propagules proliferated on the surface of fresh minimal M medium when sub-cultured without any root segment. On repeated sub-culturing, these propagules did not lose their ability to grow and produced many juvenile small spore-like vesicles during the non-symbiotic phase. Although these spores were morphologically and anatomically similar to their soil borne counter parts, they were much smaller. When placed in the vicinity of a fresh hairy root on the minimal medium or a Sudan grass seedling in sand culture, the axenically produced AM fungal propagules caused root infection, but the infection characteristics were significantly different to the original culture in terms of shape (spherical vs oval) and size (20 microm vs 45 microm) of the intraradical vesicles, and absence of 'H' branches. Sudan grass seedlings inoculated with the axenically cultured fungus showed significantly (P < 0.05) higher dry weights plant'. When compared to the plants inoculated with sand cultures, the growth parameters and the percentage infection were not significantly different. However, when both sources of inocula were used together, a synergistic effect on plant growth as well as root infection was observed. PMID- 12587746 TI - Prevalence of dental caries in India--and its trends. PMID- 12587748 TI - Dental maturity as an indicator of chronological age: radiographic evaluation of dental age in 6 to 13 years children of Belgaum using Demirjian methods. AB - The study was designed to determine dental age from orthopantomograph using Demirjian method and to investigate applicability of Demirjian method for estimation of chronological age in children of Belgaum. The sample for the study consisted of 197 subjects between 6-13 years of age. When Demirjian method was applied to Belgaum children, mean difference between true and assessed age for males showed overestimation of 0.14 years (51 days) and females showed overestimation of 0.04 years (15 days). Demirjian method showed high accuracy when applied to Belgaum children. PMID- 12587747 TI - Comparison of resin-dentine interface in primary and permanent teeth for three different durations of dentine etching. AB - Hybridization of dentine is identified as a basic bonding mechanism between resin and dentine, hence studying the micro morphology of hybrid layer may explain lower bond strength that has been observed in primary teeth. The aim of the present study was to compare the thickness and micromorphology of Hybrid layer formed in primary and permanent teeth for 3 different durations of dentine etching time (5, 10 and 15 sec) with 10% phosphoric acid as etchant. Thickness and micromorphology of hybrid layer was observed on SEM photographs obtained at 750X. Results indicated that primary teeth presented with a significantly thicker hybrid layer compared to permanent teeth at 10 sec (P<0.05) and 15 sec (P<0.01) of dentine etching. It was concluded that owing to the high reactivity of primary teeth to etchants, the etchants be applied for shorter period on primary dentine compared to permanent dentine for better performance of composite restoration in primary dentition. PMID- 12587749 TI - Prevalence of dental caries and treatment needs among children of Cuttack (Orissa). AB - An epidemiological investigation was carried out to know the prevalence of Dental Caries amongst 1257 children in the age group of 5, 8, 11 & 15 years respectively attending schools in the city of Cuttack, Orissa. The examination was carried out under natural light and dental caries was diagnosed according to W.H.O. Criteria 1983. The point prevalence of dental caries was recorded to be 64.3% with an average DMFT of 2.38. The prevalence of caries showed a pattern of occurrence i.e. prevalence consistantly increased from 5 years to 8 years age group and subsequently decreased at 11 years and 15 years age. Regarding treatment needs, 63.6% children required dental treatment for various reason and it is in accordance with dental caries prevalence of different age group. PMID- 12587750 TI - A comparison of oral hygiene status and dental caries in children on long term liquid oral medications to those not administered with such medications. AB - Young chronically ill children receive a greater sugar load from liquid medications than do healthy children. They receive variety of oral liquid medications that healthy children do not This study was planned to know the levels of oral hygiene and Dental caries in children on long term liquid oral medicines (LOM) and to know out if any difference existed between these and children not on LOM. 51 children on LOM were compared to 54 not on LOM after equalising for age, oral hygiene and diet intake. Highly significant difference was found for dmft and dmfs in 2-6 year age group and dmft+ DMFT for 6-13 years age group, mostly posterior teeth were affected and for this the difference was statistically significant in 2-6 year old children. Percentage of children with dmft>5 significantly increased in 2-6 years old children on LOM, when compared to that of control. Percentage of various grades of lesions (relating to the severity) also increased with an increase in the duration of LOM. PMID- 12587751 TI - Dental caries and oral hygiene status of school children in Davangere related to their socio-economic levels: an epidemiological study. AB - Dental caries and periodontal disease, the most commonly seen disease show striking geographic variation, socio-economic patterns and severity of distribution all over the world. Hence, an attempt has been made to determine the relationship of oral health status with socio-economic status in Davangere town. A total of 2007 children of 13 to 14 years age belonging to both sexes were examined. Type III examination was carried out during the survey. DMFcaries Index and Oral Hygiene Index was used to assess caries experience and oral hygiene status. Prasad's classification was used to know the social classification of the children. It was concluded that dental caries experience and oral hygiene status of children are strongly correlated to socio-economic status. PMID- 12587752 TI - A comparative study of microleakage below cemento-enamel junction using light cure and chemically cured glass ionomer cement liners. AB - Posterior composite resins have gained considerable importance over last few years. However, improper placement techniques continue to be a prime cause of failure of these restorations. This study was done to compare the microleakage at the tooth restoration interface extending below cementoenamel junction when posterior composite restorations were placed directly in the cavity or when placed over a glass ionomer cement liner, both chemically cured and light cured. The results have shown that light cured glass ionomer when used as a liner under a composite restoration used below gingival cementum/dentin margin has the best sealing capability. PMID- 12587753 TI - "Cariogenic potential of homemade weaning diet v/s commercial formula feeds" an in-vitro study. AB - The universal phenomenon of introducing weaning diet to the growing child sets its onset by the age of 4-6 months. This preliminary diet of the child governs his or her dental health in the long run. The main objective of this study is to investigate the histopathological and clinical invasion of deciduous teeth exposed to both the homemade weaning diet and commercial formula feeds. The study also aims at motivating the consumption of the most appropriate diet for the weaning baby. Extracted deciduous teeth were incubated with the test solutions of homemade diet and commercial formula feeds for the period of 6 weeks. Fewer carious invasion both clinically and histopathologically were seen in homemade feeds. Thus, homemade diet can be a strong candidate for the status of 'THE IDEAL WEANING FOOD'. PMID- 12587754 TI - Supplemental premolars in a 13 year old child--a case report. AB - A case of 13 old year is presented with supernumerary maxillary premolars one on either side. Radiograph revealed presence of five more supernumerary teeth unerupted in both maxilla and mandible. The implications of early diagnosis and management to prevent further complications are discussed. PMID- 12587755 TI - Streptococcal antibodies among rural school children in Bangladesh. AB - The upper limit of normal values of group A streptococcal antibodies should be known for a population concerned because it is influenced by many local conditions. As yet the reference value of the these antibodies has not been determined by using a quantitative method among Bangladeshi children. We determined the reference value of anti-streptolysin O and anti-deoxyribonuclease B among 361 apparently healthy rural Bangladeshi primary school children (aged 5 to 14 years, mean 9.2 years). Anti-streptolysin O was measured by an auto analyzer and antideoxyribonuclease B was measured by microtitre method. The geometric mean titres for the entire group was 241 IU/ml and 222 IU/ml for anti streptolysin O and anti-deoxyribonuclease B respectively. The upper limit of normal values (80th percentile) was 390 and 340 for anti-streptolysin O and anti deoxyribonuclease B, respectively. These limits should be of value to physicians, epidemiologists and clinical laboratory personnel as well. PMID- 12587756 TI - Prevalence of hypertension in Bangladesh: effect of socioeconomic risk factor on difference between rural and urban community. AB - Several studies reported that migrant Bangladeshis had greater risk for hypertension, diabetes and coronary heart disease compared with the Europeans and other migrant South Asians. So far, there has been no such study among the native population. This paper reports the hypertension prevalence and related risks among native Bangladeshis. A total of 2,361 subjects over 20 years of age were investigated. Overall prevalence rates of systolic and diastolic hypertension in the study population were 14.4 and 9.1 percent respectively. The prevalence of systolic hypertension was significantly higher in rural than in urban participants (P < 0.001). Compared with the poor the rich class had significantly higher prevalence of both systolic (P = 0.002) and diastolic (P = 0.041) hypertension. With increase of age, body mass index (BMI) and blood glucose level were significantly related to hypertension (P < 0.0001); whereas the trend for increasing waist-to-hip ratio (WHR), adjusting for social class, was not significant. Regression analysis showed that age, BMI, rural area and rich class were the strong predictors for hypertension. This study explored that hypertension prevalence in the native Bangladeshis is almost comparable to that of other Asian populations and South Asian migrants. PMID- 12587757 TI - Prognostic factors analysis in a therapeutic trail of acute lymphoblastic leukaemia (ALL). AB - It is common practice in therapeutic trials in Acute Lymphoblastic Leukaemia (ALL) to analyse the impact of prognostic factors on clinical outcome. In this study, 30 newly diagnosed ALL patients were enrolled and the median age was 14.5 years. 22 patients were male and 8 patients were female. Prognostic factors analysed include age, sex, lymphomatous presentations (bulky peripheral lymphadenopathy, mediastinal mass and enlargement of spleen and liver), initial leucocyte count, blast cell morphology and bone marrow response to chemotherapy. Remission induction and maintenance therapy with conventional combination chemotherapy and CNS prophylaxis with intra thecal methotrexate and radiotherapy were instituted to all patients for long term event free survival. Results of induction therapy and overall outcomes of treatment were observed. The impacts of analyzed prognostic factors on clinical outcomes were assessed. 3 patients (10%) failed to go into remission after 12 weeks of therapy. 27 patients (90%) obtained remission. Among 27 patients those obtained remission, 17 patients continued event (failure to remit, relapse and death) free survival, 7 patients suffered relapse of their leukaemia and 3 patients died during maintenance therapy. Events were observed more among patients age group > or = 20 yrs (84%), initial WBC count > or = 50 x 10(9)/L (58%) and FABL2 morphology (80%). PMID- 12587758 TI - Incidence and prognostic significance of right bundle branch block complicating acute myocardial infarction. AB - The incidence and hospital courseofacute myocardial infarction wereobserved among randomly selected 560 patients. The mean age of the patients was 53 +/- 6.67 years with male-female ratio 2.6:1. The incidence of right bundle branch block (RBBB) was 15% (84 out of 560 patients). New RBBB, old RBBB and RBBB of indeterminate age were observed in 33 (39.25%), 23 (27.40%) and 28 (33.25%) cases respectively. RBBB was isolated in 50 (60%) cases and bi-fascicular in remaining 34 (40%) cases. The rate of use of thrombolytic was greater in RBBB group than non-RBBB group (52% vs 30.67%, P < 0.05). The complications were more frequently observed in patients with RBBB: in-hospital mortality, 27.40% vs 10.90% (P < 0.01); use of temporary pacemaker, 15% vs 9% (P < 0.05); and heart failure, 50% vs 35% (P < 0.05). In sub-group analysis, in-hospital mortality rate was higher among bi-fascicular group than isolated RBBB group (P < 0.05). Thus it appeared from the results that the incidence of RBBB is relatively common i.e. 15% over all (84 out of 560) and 5.90% new (33 out of 560) and despite greater use of thrombolytics, some specific in-hospital complications were significantly higher in patients with RBBB than without, RBBB. PMID- 12587759 TI - Comparative study between tubotympanic and atticoantral types of chronic suppurative otitis media. AB - This prospective study was done to compare the different aspects of tubotympanic (group I) and atticoantral types (group II) of chronic suppurative otitis media. The study included 200 patients of chronic suppurative otitis media, of them 100 were of tubotympanic type and 100 were with atticoantral disease. The age (mean +/- SD) of group I and II were 17 +/- 2.7 years and 12 +/- 2.3 years respectively (P < 0.05). The majority of the patients having poor socio-economic condition 69% in group I and 87% in-group II. In both groups, slum dwellers with bathing habit in pond or river and illiterate were more sufferer (P < 0.05). There was central perforation in all patients in-group I, and in-group II, 67% were with attic and 33% were with marginal perforations. In group I, aural discharge was mucoid or mucopurulent in majority of cases 80% and in-group II foul smelling scanty ear discharge was from 88% of subjects. In atticoantral type, hearing impairment was more than tubotympanic disease. Complications were more in atticoantral type of disease. In-group II, post auricular sinus 25% was the commonest extracranial complication followed by subperiosteal abscess 18%, and meningitis 10% was the commonest intracranial complication followed by brain abscess 8%. In tubotympanic type conservative treatment with reconstructive surgery were the treatment of choice, whereas radical or some modifications of radical surgery (with or without reconstruction) were the treatment plan for atticoantral disease. Among the groups the mortality rate was only 4% in-group II and was due to intracranial complications. The study recommends early detection and effective surgical treatment of the disease to improve the situation. PMID- 12587760 TI - Verbal autopsy for maternal death. AB - This cross sectional study was done in Gopalganj district from September 1997 to August 1998. To ascertain the magnitude and causes of maternal deaths and perinatal and neonatal outcome of the babies. Two hundred five in maternal deaths occurring from 1994 to 1997 were investigated by verbal autopsy. Teenager mothers had more deaths in first pregnancy. Maternal deaths increased with gravidity above 30 years. Haemorrhage and eclampsia were common causes of death. Eclampsia was prominent in primigravids <20 years while haemorrhage occurred more in multigravidas between 25-35 years of age (P = 0.029 for age, P = <0.001 for gravidity). Nearly 64% of maternal deaths occurred in postpartum period. Among live births, 45.6% babies died before their first birth day. The findings support the need of increasing the age of marriage and first childbirth, through strengthened family planning programme and ensuring skilled birth attendance during delivery. It also stresses need for communication on danger signs of pregnancy and childbirth. PMID- 12587761 TI - Is evidence-based practice compatible with best practice? PMID- 12587762 TI - Current concepts in the management of oral squamous cell carcinoma. AB - This paper focuses on how patients with oral squamous cell carcinoma (SCC) are assessed and managed once they have been referred for specialist care. Current and future treatment options are presented and basic management principles are discussed. It is our hope that this will enable the general dental practitioner to gain insight into the process and reasoning behind the multidisciplinary treatment of these complex cases. PMID- 12587763 TI - Oral lichen planus: causes, diagnosis and management. AB - Oral lichen planus (OLP) is a chronic inflammatory disease of unknown etiology. In this paper we review the clinical and histological features of OLP, process of OLP diagnosis, causes of OLP, management of OLP patients and medical treatment of OLP lesions. Approximately 0.2 per cent OLP patients develop intra-oral carcinoma each year compared with approximately 0.005 per cent Australian adults. Possible mechanisms of increased oral cancer risk in OLP patients are presented. The aims of current OLP therapy are to eliminate mucosal erythema and ulceration, alleviate symptoms and reduce the risk of oral cancer. Patient education may improve the outcomes of OLP therapy and further reduce the risk of oral cancer in OLP patients. Although OLP may be diagnosed clinically, appropriate specialist referral is required for: (i) histological diagnosis; (ii) assessment of causative/exacerbating factors, associated diseases and oral cancer risk; (iii) patient education and management; (iv) medical treatment; and (v) long-term review and re-biopsy as required. PMID- 12587764 TI - Dental erosion in asthma: a case-control study from south east Queensland. AB - BACKGROUND: Asthma medication places patients at risk of dental erosion by reducing salivary protection against extrinsic or intrinsic acids. But patterns of lesions in asthmatics may differ from patterns in non-asthmatics, because gastro-oesophageal reflux (GOR) is found in 60 per cent of asthmatics. METHODS: The lesions in 44 asthma cases were compared to those of age and sex match controls with no history of asthma or medications drawn from the dental records of 423 patients referred concerning excessive tooth wear. The subjects were 70 males age range 15 to 55 years and 18 females age range 18 to 45. Anamnestic clinical data were compared between the two groups. Models of all 88 subjects were examined by light microscopy, and wear patterns were recorded on permanent central incisor, canine, premolar and first molar teeth. RESULTS: Clinical differences were a higher incidence of tooth hypersensitivity, xerostomia, salivary gland abnormalities, gastric complaints, and self induced vomiting in the cases. No differences were found between the cases and controls on citrus fruit and acid soft drink consumption. More occlusal erosion sites were found in cases, whereas more attrition sites were found in the controls. There were no significant differences in palatal erosion on maxillary anterior teeth found between cases and controls. Lingual erosion of the mandibular incisors, found only in GOR patients, was not observed. CONCLUSIONS: A higher incidence of erosion was found in asthmatics. Gastro-oesophageal reflux symptoms were not associated with the sign of lingual mandibular incisor erosion. The clinical significance is that asthmatics are at risk of dental erosion from extrinsic acid, but GOR does not appear to contribute in a site-specific manner. PMID- 12587765 TI - Studies on dental erosion: An in vivo-in vitro model of endogenous dental erosion -its application to testing protection by fluoride gel application. AB - BACKGROUND: The objective in this study was to develop an in vivo-in vitro model of endogenous erosion, with a view to exploring the potential for some degree of its control by the use of topical fluoride gel application to teeth. METHODS: Six volunteers each wore a small clasp retained palatal acrylic appliance to which six sterilized enamel tiles were bonded. Three tiles on each appliance were coated extra-orally with either 1.23 per cent acidulated phosphate fluoride (APF) or 2.2 per cent sodium fluoride (NaF) neutral gel for four minutes prior to multiple periods of exposure to the simulated gastric acid, cumulating in 16, 36, 80 and 150 minutes of exposure. Impressions of the enamel tiles prior to and following acid exposure permitted dies to be prepared. These were sectioned through the exposed areas and examined under a stereomicroscope to assess maximum depths of erosion. RESULTS: The depth of erosive demineralization of enamel was found to be greatly reduced with increased frequency of APF gel application. The reduction in enamel loss was less following topical application of NaF gel. CONCLUSIONS: It was concluded that fluoride gels significantly reduced enamel erosion using this in vivo-in vitro model and therefore, if prescribed appropriately, should help reduce tooth tissue loss from endogenous erosion. PMID- 12587766 TI - Patterns of referral of patients undergoing surgical management for oral cancer. AB - BACKGROUND: Oral cancer is most often diagnosed by dentists or dental specialists. We were interested in analyzing demographic and referral data for a cohort of patients referred for the management of oral cancer to our unit. METHODS: A consecutive review of all patients treated by oral and maxillofacial surgery at the Austin and Repatriation Medical Centre over the last 10 years. Inclusion criteria included those patients with malignant oral and perioral disease that underwent surgical management. Exclusion criteria included those patients who did not have malignant oral disease, who did not have surgical management and those patients who were not treated by our unit as the primary surgeon. RESULTS: A total of 113 patients were identified. The mean age for presentation for the group was 61.6 years and the male to female ratio was 1.2:1. Approximately 70 per cent of patients were smokers or had smoked in the past. Over the last 10 years there has been a steady increase in referrals to our unit. Over half of all patients referred were from the Royal Dental Hospital of Melbourne while the majority of the remainder of the referrals were from private oral and maxillofacial surgeons. A significantly increased number of referrals from other sources were identified. Surveillance techniques that can be used in general dental practice are also described. CONCLUSIONS: A high proportion of referrals from dental sources highlights the importance of the dentist and dental specialist in the diagnosis and referral of patients with suspected oral cancer. Patients can either be referred to a dental hospital, private oral and maxillofacial surgeons or directly to a public hospital oral and maxillofacial surgery unit treating oral cancer, such as the one at the Austin and Repatriation Medical Centre. PMID- 12587767 TI - Caries inhibitory effect of fluoridated sugar in a trial in Indonesia. AB - BACKGROUND: In some regional areas of Indonesia, caries prevalence is increasing rapidly. As water, salt or milk fluoridation were not considered suitable for use throughout Indonesia, and fluoridated tooth paste is mostly too expensive, a fluoride co-crystallised sugar containing 10 ppm fluoride was prepared. Its efficacy in inhibiting caries development was tested in a field trial. METHODS: The field trial was established in Medan, Sumatera. All dietary background data necessary to ensure the safety of a trial were collected. Subjects chosen were 176 children who were residents of two orphanages and a boarding school for children of poor rural families. The trial used a double-blind format. Close monitoring of fluoride consumption was maintained, and fluoride excretion rates were assessed six monthly by urinary fluoride analysis. RESULTS: Records of total tooth surface caries present initially and after 18 months of sugar supply showed that the children using fluoridated sugar had significantly fewer carious lesions than those who used normal sugar. CONCLUSION: This result indicates that sugar might be considered as a further vehicle for supplementary dietary fluoride in communities where there is a high caries prevalence or high caries risk and little exposure to fluoride. PMID- 12587768 TI - Dental screening of older adults living in residential aged care facilities in Perth. AB - BACKGROUND: This study aimed to estimate the dental treatment needs and oral health status of a sample of older adults in residential aged care facilities in Perth. METHODS: The 348 participants (> or = 65 years) were interviewed and screened in 25 facilities. The screenings were carried out by one examiner using a mirror and a portable light. RESULTS: Over half (52 per cent) of the participants were edentulous and 45 per cent of those edentulous participants for whom a recording was made (n=174) had oral mucosal conditions. The 164 dentate participants had a mean of 12 disease-free standing teeth, a mean decayed, missing or filled teeth (DMFT) of 24.7 (mean DT 0.8, mean FT 5.3) and half of them required the removal of supragingival calculus. Of those with root caries experience (n=127), a mean of 1.3 untreated decayed roots and a mean of 1.9 roots covered in plaque were recorded. The majority of the participants (83 per cent) were pensioners eligible for government subsidized dental care and 47 per cent were reported by the Directors of Nursing to have dementia. CONCLUSIONS: The data collected here demonstrate poor oral health conditions and a substantial treatment need in a neglected population. More people in nursing homes and hostels are keeping their natural teeth compared with a similar population studied 13 years ago. PMID- 12587769 TI - A study of the comparative efficacy of three common analgesics in the control of pain after third molar surgery under local anaesthesia. AB - BACKGROUND: The aim of this study was to evaluate the comparative efficacy of three commonly used analgesics (Panadeine, Diflunisal and Etodolac) in the control of pain after third molar surgery under local anaesthesia. METHODS: A randomized control study. Outcome of primary efficacy was judged by overall assessment of the area under the curve of graphs for pain intensity, measured from serial visual analogue scales over a 24-hour period. Other measures of efficacy included the number (per cent) of patients who took 'additional' analgesics and the incidence of adverse effects occurring in each treatment group over the study period. RESULTS: The three drugs were effective in the control of post-operative pain (p<0.01). Variations in pain intensity and the use of additional medication between the treatment groups were observed over the study period. The Diflunisal group experienced less pain than the Panadeine or Etodolac group (p<0.01). Furthermore, a lesser number of those in the Diflunisal group used additional medication compared to the other two groups (p<0.01). The incidence of side effects from all three drugs was low. CONCLUSION: Diflunisal is superior in the control of pain following third molar surgery under local anaesthesia than either Panadeine or Etodolac, and has few side effects. PMID- 12587770 TI - Dental caries experience of preschool children from the North Brisbane region. AB - BACKGROUND: To report the dental caries experience of preschool children within the north Brisbane region and to investigate the association between selected social and demographic variables and disease presence. METHODS: A cross-sectional sample of 2515 children aged four to six years was examined in a preschool setting using decayed, missing, filled teeth/surface (dmft/dmfs) and percentage caries free indices. A self-administered questionnaire obtained information regarding selected social and demographic variables. The data were analysed using the chi-square and one way analysis of variance procedures at the 5 per cent level of significance. RESULTS: Overall, 1668 (66.3 per cent) children were disease free at the examination and mean dmft was 1.4+/-2.77 and dmfs was 2.28+/ 6.00. Dental caries occurred more frequently and severely in children from non Caucasian background (p<0.000), family language other than English (p=0.001) and lower socio-economic status (p<0.000). CONCLUSIONS: Significant associations between child ethnicity, language spoken at home, socio-economic status and caries presence have been identified. Epidemiological data can be used for improved public oral health service planning and resource allocation within the region. PMID- 12587771 TI - Resin modified glass-ionomers: strength, cure depth and translucency. AB - BACKGROUND: According to the classification suggested by McLean, Wilson and Nicholson only three materials, currently marketed, fit the designation of resin modified glass-ionomers. These materials will undergo an acid/base setting reaction but also contain a limited quantity of a monomer that will polymerize as a result of irradiation. The quantity of polymer is limited to the extent that it will not interfere with the normal acid/base setting reaction and will therefore allow for the ion exchange adhesion with tooth structure that is typical of glass ionomer. There is a third setting reaction incorporated to ensure remaining monomer, that is not affected by irradiation, will still polymerize. A series of experiments were carried out to determine the effect of the three types of setting reaction on the strength, depth of cure and translucency of these three materials. METHODS: Specimens, both with and without irradiation, were subjected to a shear punch strength test. To determine the depth of cure brought about through irradiation, specimens were tested immediately after construction according to the test in ISO - 4049 2000 (E). Translucency is clinically significant and will vary according to whether the material has been subjected to irradiation or cured through the acid/base reaction alone. RESULTS: The strength of all materials tested was higher in specimens subject to irradiation. The depth of cure was found to be both shade and irradiation time dependent. Irradiated specimens were found to be only marginally more translucent than those allowed to set without irradiation. CONCLUSIONS: It was concluded that, for cavities more than 3 mm deep, these materials should be placed incrementally to allow for a full irradiation initiated cure. PMID- 12587772 TI - Foreign bodies in the maxillary antrum: a case report. AB - A case is presented where six gutta percha (GP) points were introduced into the right maxillary antrum during routine endodontic treatment on an upper second premolar tooth in a poorly controlled insulin-dependent diabetic. The surgical management of the case is described and the possibly more serious sequelae are discussed. Comments are made on the appropriateness of the treatment plan. PMID- 12587773 TI - Submandibular gland aplasia and progressive dental caries: a case report. AB - When severe caries occurs in mandibular permanent incisor teeth, the clinician should consider the possibility of associated submandibular gland aplasia or salivary hypofunction. Early diagnosis of submandibular gland disease is essential, as operative problems involving restoration of mandibular incisor teeth are considerable. Furthermore, progressive severe dental caries can present a dilemma for the clinician in affected individuals, despite intensive preventive and restorative therapy. A case report describing severe progressive dental caries and enamel demineralization of the permanent mandibular incisor teeth in a young girl is presented. Further investigation revealed absence of functional bilateral submandibular salivary glands contributing to the rapid breakdown of the teeth despite intensive preventive measures. PMID- 12587774 TI - Defibrillation of a patient/restoration of a single tooth. PMID- 12587775 TI - A radiographic assessment of the prevalence of pulp stones. PMID- 12587776 TI - Multiple doses of an antibiotic over a series of dental appointments--the problem. Part 1. PMID- 12587777 TI - Multiple doses of an antibiotic over a series of dental appointments--the problem. Part 2. PMID- 12587778 TI - Immunosuppressive effect of polycyclic aromatic hydrocarbons by induction of apoptosis of pre-B lymphocytes of bone marrow. AB - Polycyclic aromatic hydrocarbons (PAH) are ubiquitous environmental pollutants, distinguished by genotoxic, hepatotoxic, nephrotoxic and immunotoxic effects. Especially secondary toxicity after bioactivation by microsomal monooxygenases (dependent on cytochromes P450) is characteristic of them. The immunotoxic effect is the result of very global impact on immunological reactivity of an organism and immunosuppression by induction of apoptosis of pre-B lymphocytes represents one of its particular forms. It has been proved that the effect of PAH is caused mostly by the following mechanisms: enzymatic induction by the way of activation of AhR (Aromatic hydrocarbon Receptor); alteration of cellular DNA; development of oxidative stress; increase in the concentration of intercellular calcium and decline of activity of NF-kappaB (Nuclear Factor-kappa B). Most sensitive to these changes are particularly B-lymphocytic precursors and pre-B lymphocytes. Intensity of entire manifestations is also considerably dependent on the presence and intensity of mechanisms of active or passive resistance of cells. PMID- 12587779 TI - Microelements and inherited metabolic diseases. AB - In addition to the main groups of inherited metabolic diseases, including mitochondrial, peroxisomal and lysosomal defects, organic acidurias, porphyrias, defects of amino acids, saccharides and fatty acids metabolism, disorders of transport and utilisation of microelements have also been recognized. Recent findings concerning hereditary hemochromatosis (iron), Wilson and Menkes diseases (copper), molybdenum cofactor deficiency (molybdenum), defects of cobalamine synthesis (cobalt) and acrodermatitis enteropathica (zinc) are reviewed. PMID- 12587780 TI - Time dependent appearance of selected apoptotic markers and usefulness of their detection in vitro. AB - Many experiments have demonstrated that some cell lines are resistant to chemically induced apoptosis in vitro, and that apoptosis itself is far from being a homogenous phenomenon. Here we show that 10 microg/ml etoposide elicited only minor changes in Bowes human melanoma cells (temporary decrease in cell viability and proliferation, transient phospatidylserine externalization and caspase-3 activation), which weren't clearly capable to start apoptotic pathway in the entire treated population. On the other hand, potassium chromate at concentration of 150 microg/ml executed cell death bearing some features of apoptosis (cell blebbing, caspase-3 activation and cytoskeletal changes) but lacking or showing weakly others (DNA fragmentation and phospatidylserine externalization). Our results suggest that in detecting apoptosis several fault proof detection systems are to be used to avoid misleading results and conclusions in each experimental setting. PMID- 12587781 TI - Antiacetylcholinesterase activity of cyclosporine--a comparison of single and repeated administration and effect of 7-methoxytacrine. AB - The aim of this work is a comparison of single and repeated peroral administration of cyclosporine (CsA) and the interaction of repeated administration of CsA and 7-methoxytacrine (MEOTA) on the activity of acetylcholinesterase (AChE) in the frontal cortex, hippocampus, septum, and basal ganglia in rats. Both single and repeated administration of CsA diminished the activity of AChE in the frontal cortex, septum and basal ganglia, while the enzyme activity in the hippocampus was diminished only in the case of repeated CsA, as well as repeated CsA + MEOTA administration. Repeated administration of CsA led to a further augmentation of anticholinesterase activity only in the frontal cortex and--in a lesser extent--in the basal ganglia. No augmentation of AChE activity was observed in the hippocampus and septum. PMID- 12587782 TI - The impairment of spatial memory following low-level sarin inhalation exposure and antidotal treatment in rats. AB - 1. To study the influence of antidotes on low-level sarin-induced impairment of cognitive functions, the rats were exposed to three various low concentrations of sarin (LEVEL 1-3) for 60 minutes in the inhalation chamber. In addition, one group of rats was exposed to LEVEL 2 of sarin repeatedly. 2. Testing of cognitive functions was carried out using the Y-maze evaluating learning and spatial memory. The correct averse behavior of sarin-exposed rats in the Y-maze was tested several times within four weeks following sarin inhalation exposure and antidotal treatment to look for any cognitive impairments. 3. The results were compared to the Y-maze performance of sarin-exposed rats without antidotal treatment and control rats exposed to pure air instead of sarin with or without antidotal treatment. While antidotal treatment was able to eliminate a short-term deficiency in the Y-maze performance in rats exposed to the LEVEL 1 of sarin, a significant decrease in the Y-maze performance in rats exposed to sarin at the LEVEL 2 and 3 was only shortened. Sarin-induced spatial memory impairments in rats exposed repeatedly to sarin at the LEVEL 2 was also shortened when rats were treated following each sarin inhalation exposure. 4. The findings confirm that antidotes currently used for nerve agent poisonings are beneficial for the treatment of rats singly or repeatedly exposed to non-convulsive symptomatic or even clinically asymptomatic concentrations of sarin. PMID- 12587783 TI - Diagnostic value of C-reactive protein in patients with angiographically documented coronary heart disease. AB - AIM: The aim of this study was to evaluate the diagnostic value of serum C reactive protein (CRP) level measurement in predicting coronary artery disease (CAD) that can be shown angiographically. METHODS: CRP levels were determined in the blood of 198 patients (patients group, PG) with angiographically documented coronary artery disease and compared with that of 85 patients (control group, CG) who had a clinical indication for coronary angiography but have no angiographically determined coronary artery stenosis, as well as with that of 41 healthy volunteers as a healthy control group (HG) who did not have any complaint and did not have coronary angiography. CRP levels were measured 24 hours prior to angiography in PG and CG patients, and in the morning after not having eaten for same time. Any coronary artery stenosis or plaque formation was defined as CAD. Severity of the disease was assessed by both the number of diseased vessels (0 to 3) and the degree of stenosis (<50% mild, 50-70% moderate and >70% severe). RESULTS: Receiver Operating Characteristics (ROC) curves of CRP in angiographically documented CAD group showed a diagnostic value of 0.659 in female patients, followed by 0.542 in male patients, in predicting CAD. CRP levels were found to be significantly different between groups, higher in PG (6.2 +/- 0.86 mg/L) than those of CG (3.7 +/- 0.92 mg/L) and HG (0.854 +/- 0.2 mg/L) (p<0.05). CRP levels were not associated with the number of diseased vessels, neither with the degree of the occlusion (p>0.05). Multiple logistic regression analysis after adjustment for the established coronary risk factors showed CRP as an independent discriminating risk factor for CAD. CONCLUSION: It is concluded that CRP measurement has a value in predicting the presence of angiographically documented CAD. However, CRP levels were not associated with the degree or severity of CAD. PMID- 12587784 TI - Some recruitment aspects of population photoscreening of amblyogenic factors at children younger one year. AB - The aim of the study was to evaluate the keystone role of paediatric general practitioners (PGPs) in our system of photoscreening of amblyogenic factors at children younger one year. The parental involvement on the participation of their children at photoscreening was also analysed. From June 2000 to February 2001 we have performed 780 photoscreening tests of children aged from 3 months to 31 months (mean age 9.7 months). The photoscreening test was voluntary. Parents was invited to visit screening center by PGP's recommendation and by offering the Invitation/information brochure. The brochure was distributed by 56 paediatric general practitioners during obligatory examinations at 5., 8. resp. 12. month of age from June to November 2000. Based on the questionnaire (return rate 89%) we documented following data: the number of children of particular age in the care of PGPs was 2060, 1458 Invitation/information brochures was distributed by PGPs. PGPs attended for more children at particular age distributed relatively less number of brochures if compared to PGPs with lower number of relative children. The willingness to the photoscreening programe participance of parents of children belonging to PGPs attended our educational seminary exhibited independence on distance between PGP office and our department in the strong contrast to families belonging with PGPs not participating on educational programme. Our further effort on elevating the percentage of children population participation on screening must be focused on education of PGP with large number of children at particular age and on ensuring the issuing of brochures or other forms of invitation at PGPs' offices. PMID- 12587785 TI - Maxillary sinus augmentation with deproteinized bovine bone (Bio-Oss) and Impladent dental implant system. Part II. Evaluation of deprotienized bovine bone (Bio-Oss) and implant surface. AB - The objective of this clinical study was to determine the predictability of endosseous implant placed in a maxillary sinus augmented with deproteinized bovine bone (Bio-Oss). A total of 185 implants (109 titanium and 76 hydroxyapatite-coated) were placed in 77 patients representing 92 sinuses either a one- or two-stage surgical technique. A mixture of venous patient's blood and Bio-Oss was used alone within 20 sinuses (Group 1), or in combination with autogenous bone within 72 sinuses (Group 2). Thirty-nine implants were placed in Group 1 and 147 implants were inserted in Group 2. The grafted sinuses were evaluated clinically and radiographically at second stage surgery. According to certain criteria, of the implants placed, only two titanium implants (1.08%) failed with 98.91% implant survival. There was no statistically variable difference for the use of hydroxyapatite-coated or titanium implants. The two failed implants were from Group 2. No clinical benefit has been achieved from the combination with autogenous bone (P<0.05). All the grafted sinuses were sufficient to place dental implants of at least 12 mm length (100% graft success). The results of this short-term study support the hypothesis that Bio Oss can be a suitable material for sinus augmentation. PMID- 12587786 TI - Evaluation of the accuracy of panoramic radiograph in the definition of maxillary sinus septa. AB - The purpose of this study was to determine the accuracy of panoramic radiography in identification of maxillary sinus septa. Out of 68 sinuses were radiographically examined using both panoramic and computerized tomographic radiographs (CT scan). Using CT scan, 24 (35.9%) out of 68 cases maxillae showed at least one septum, 22 sinuses (32.3%) showed one septum, whereas two sinuses (2.9%) exhibited two septa. Panoramic radiograph led to a false diagnosis regarding the presence or absence of sinus septa in 18 of 68 sinuses (26.5%). On the other hand, they gave negative diagnosis of sinus septa in 12 of 24 septa (50%). There was fully agreement between the two methods (positive septa) only in 12 of 24 septa (50%). We cannot depend on panoramic radiograph for the detection of sinus septa because it can lead to false or negative results. PMID- 12587787 TI - Low dose ketoconazole therapy and thyroid functions in rats. AB - To determine whether low dose ketoconazole (KTZ) has antithyroid action, we studied thyroid function tests in the 13 rats treated with KTZ (20 mg/kg twice daily) for thirty days. Serum triiodothyronine and thyroxine levels were decreased (P<0.05) and serum thyroid-stimulating hormone levels were mildly increased (P>0.05) at the end of treatment. Histopathological analysis of the thyroid glands demonstrated an increase in cylindrical cells in study group, but the epithelial cells were mainly cubical in control group. These findings showed that low dose KTZ had antithyroid effect in rats. The responsible mechanisms may be direct effect of the drug on thyroid gland. PMID- 12587788 TI - Yellow nail syndrome or diffuse lymphatic network disease. AB - We report a man aged 68 years old with pneumothorax and chronic bilateral pleural effusion in association with a history of yellow nails. The diagnosis of yellow nail syndrome based on yellow nails, lymphedema, chronic pleural effusion and intestinal lymphangiectasia. PMID- 12587789 TI - Mitochondrial DNA as a clonal tumor cell marker: gliomatosis cerebri. AB - The aim of this study was a clonal analysis of gliomatosis cerebri (GC), a rare disease characterized by diffuse, extensively infiltrating glial tumors of the central nervous system. Two females of the series were not informative in assays for X-chromosomal inactivation, and a polycytosine tract of the mitochondrial DNA (mtDNA) was tested as a clonal marker. Following fluorescent PCR, a fraction of human individuals shows several electrophoretic bands in normal tissues, some of which can be lost in corresponding glial tumors. Two male patients of our series fulfilled this prerequisite and were thus informative. In patient 1, four tumor samples from the left temporal and occipital cortex, histologically corresponding to WHO grades III and IV, showed an identical loss of bands, which was not observed in tumor-free brain and in tumors from the left cerebellum, from fornix and corpus callosum, and from the right occipital cortex, corresponding to WHO grades III and IV. Since this patient exhibited a TP53 mutation in exon 7, we sequenced this exon in all tissue samples of this individual. The mutation was found selectively in the tumor samples with a loss of mtDNA bands. In patient 2, all tumors (histologically corresponding to WHO grade II) from putamen, thalamus, midbrain and right parietal cortex exhibited an identical loss of bands in the mtDNA analysis. Taken together, these results support that even distant tumors in a patient with GC can share a common clonal origin. They demonstrate the extraordinary mobility and infiltrative power of these tumor cells. PMID- 12587790 TI - Adenoviral (full-length) Apo2L/TRAIL gene transfer is an ineffective treatment strategy for malignant glioma. AB - Death ligand-mediated apoptosis is a promising strategy of gene therapy for human malignant glioma. We here report that the infection of human malignant glioma cell lines with an adenoviral vector encoding full length human Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand (Ad-Apo2L/TRAIL) results in strong Apo2L/TRAIL transgene expression and the release of full-length Apo2L/TRAIL into the cell culture medium. However, Ad-Apo2L/TRAIL is a poor inducer of cell death, even in the presence of inhibitors of protein synthesis, in human glioma cell lines which are sensitive to soluble recombinant human His tagged Apo2L/TRAIL (amino acids 114-281). Moreover, Ad-Apo2L/TRAIL gene transfer inhibits soluble His-tagged Apo2L/TRAIL-induced apoptosis, strongly suggesting that the adenovirally encoded full-length Apo2L/TRAIL is not a suitable molecule for glioma cancer gene therapy. This study has important implications for the future development of therapeutic strategies aiming at death receptor activation in refractory cancers such as malignant glioma. PMID- 12587791 TI - Masson's vegetant hemangioendothelioma: case report and literature review. AB - Masson's vegetant intravascular hemangioendothelioma has only been reported intracranially in 12 patients. The pathological diagnosis is important given its benign natural history. We report the 13th case in a woman who presented with headaches and dysphasia. A thorough literature review is presented and an appropriate management strategy is proposed. PMID- 12587792 TI - Primary dural low grade BCL-2 negative follicular lymphoma: a case report. AB - We report a case of low-grade follicular lymphoma presenting as a primary lesion in the intracranial dura. Based on the preoperative MRI, our patient was presumed to have meningioma. A surgical excision was performed and the histology showed features of a grade 1 follicular lymphoma predominately small-cleaved cell type. Postoperative investigations confirmed that it was the only site of involvement. The patient then received cranial radiotherapy and she is doing well with a follow up period of 18 months. To the best of our knowledge, it is the first case of a primary dural grade 1 follicular lymphoma, small-cleaved cell type. PMID- 12587793 TI - Spinal cord hemangioblastoma: surgical strategy and clinical outcome. AB - BACKGROUND AND AIM: The authors present a retrospective analysis of 14 patients treated for spinal cord hemangioblastoma (SCH). This study was conducted to evaluate and compare the difference of functional outcomes associated with the extent of surgical removal of the tumor. METHODS: Eleven patients were male and three were female, with a mean age of 37.2 years (19-62 years). Pre- and postoperative magnetic resonance imaging was performed in all cases. Preoperative angiography was performed in 11 cases and preoperative embolization in four. The follow-up period ranged from 15 to 161 months (median, 47 months). RESULTS: Thirteen patients had intramedullary tumor, and the remaining one had an extradural lesion. All patients underwent surgical removal, and total removal (TR) was achieved in 10 cases. In four patients with preoperative embolization, intraoperative bleeding was minimal and TR was possible. In three of four patients without TR, their functional outcomes were aggravated postoperatively. At the last follow-up (15-161 months), eight patients were improved, three were stationary, and three deteriorated. All patients who showed improvements underwent TR. The statistical analysis showed that TR produced a significantly better outcome than incomplete removal (p = 0.015). CONCLUSION: TR resulted in a better outcome for SCH. Preoperative embolization could be effective in the reduction of intraoperative bleeding and facilitate TR with an improved surgical outcome. PMID- 12587794 TI - Intra-arterial carboplatin and intravenous etoposide for the treatment of metastatic brain tumors. AB - Metastatic brain tumors (MBT) are the most frequent complication of systemic cancer and often respond poorly to treatment. Median survival is only 16-24 weeks after conventional radiation therapy. Regional intra-arterial (IA) administration of chemotherapy results in increased tumor uptake of drug and may improve response rates and survival. Twenty-seven patients with MBT who had received prior irradiation were treated with IA carboplatin (200 mg/m2/d) and intravenous (i.v.) etoposide (100 mg/m2/d) for 2 days every 3-4 weeks. Eighteen patients (67%) had received prior systemic chemotherapy for their primary tumor. Patients ranged in age from 19 to 68 years (mean 48.1). Thirteen of 24 evaluable patients had objective responses (54.2%). There were 6 complete responses (25%), 6 partial responses (25%), 1 minor response (4.2%), 7 stable disease (32%), and 5 progressive disease (20.8%). Some patients with multifocal tumors had a mixture of responses. The median time to progression was 16.0 weeks overall and 30.0 weeks in responders (range 6-118 weeks). Overall median survival from the time of protocol initiation was 20.0 weeks. In six responders, death occurred due to systemic illness unrelated to MBT progression. Therapy was well tolerated, with predominantly hematologic toxicity. Angiographic complications were rare. Although these are preliminary results, IA carboplatin and IV etoposide is safe and well tolerated, appears to be active against brain metastases, and warrants further study. PMID- 12587795 TI - High-dose-rate stereotactic brachytherapy for patients with newly diagnosed glioblastoma multiformes. AB - PURPOSE: To evaluate high-dose-rate (HDR) stereotactic brachytherapy (STBT) for glioblastoma multiforme (GBM). MATERIALS AND METHODS: Between August 1994 and December 1998, 28 patients with newly diagnosed GBM underwent surgery, external beam radiotherapy (EBRT) and HDR STBT. STBT eligibility criteria included unifocal lesions, residual tumor < or = 6 cm in maximum diameter, supratentorial lesions, tumors not crossing the midline, tumors without subependymal spread and Karnofsky performance status (KPS) > 60. STBT was delivered over five consecutive days with two fractions per day for a total median dose of 30 Gy. Twenty-eight STBT eligible GBM patients treated with surgery and EBRT only over the same period were matched controls. RESULTS: Median survival times for the STBT group and controls were 19.5 versus 12.5 months; one and two year survival rates were 89% versus 42% and 61% versus 28%, respectively (p = 0.12). Using multivariate analysis, age, KPS and HDR STBT were significant factors predicting survival. By RPA class, 2-year survival rates for STBT and controls were: III--78% versus 50%; IV--40% versus 0%; V--21% versus 15%, respectively. Corresponding median survival times in months were: 41.6 versus 21.2 (p = 0.39); 16.7 versus 12.1 (p = 0.36); 18.7 versus 10.6 (p = 0.02). No major complications were found in the STBT arm. CONCLUSIONS: Because of small patient numbers, median survival time increases were only statistically significant in the RPA Class V patients, but a strong survival time trend emerged favoring patients undergoing HDR STBT. Further prospective study is warranted to fully assess the merits of this technique for GBM management. PMID- 12587796 TI - Clinicopathological study of seven cases of symptomatic supratentorial subependymoma. AB - Subependymomas are rare, slow-growing tumors, the majority of which are found incidentally at postmortem examination. The authors retrospectively analyzed seven cases of symptomatic supratentorial subependymomas. Five were females and two were males, ranging in age at operation of 6-50 years (median 45). The follow up period ranged from 1.5 to 8.3 years. Tumors were intraventricularly located as a lobulated mass with cystic changes: four in the frontal horn, two in the trigone, and one in the third ventricle. Moderate to marked enhancement was noted in two tumors of the trigone and in one tumor of the frontal horn on both CT scan and MR imaging. MR spectroscopy of a recurrent subependymoma demonstrated a higher Cho/Cr ratio of 2.66, compared with a Cho/Cr ratio (0.48) of a non recurrent subependymoma. Angiography, which was performed in four patients, revealed no staining in two and delayed modest staining in two. Radiosurgery was performed in two patients but was ineffective. Five patients with gross total tumor resection showed no evidence of tumor recurrence to the last follow-up. The two subtotally resected trigonal tumors progressed two years after operation. No histological difference except MIB-1 index was noted between recurrent and non recurrent cases. In conclusion, we suggest that subependymoma could show progressive biological behavior, especially in cases of markedly enhancing, irregularly contoured, large tumors located in the trigone. For symptomatic supratentorial subependymomas, gross total resection is the treatment of choice and radiation has little effect on tumor control. PMID- 12587797 TI - Dural cerebellopontine angle metastasis from malignant parotid oncocytoma. AB - Malignant oncocytoma of the parotid gland is a quite rare tumor, with only 40 cases with unequivocal histological diagnosis reported in the literature. No cases with intracranial metastasis have been described. The authors report a very unusual case of malignant parotid gland oncocytoma with a large dural extracerebellar metastasis occurring in the contiguous cerebellopontine angle six months after surgery for the primary tumor. Only a partial removal of the dural metastasis was possible because of the cranial nerve and vessel encasement within the mass. Surgery and irradiation resulted in one-year survival. Interestingly, no destruction and infiltration of the petrous and temporal bones were found both at radiological and surgical exploration. We may suggest that metastatic spread from the parotid region to the cerebellopontine angle occurred through the mastoid cells or by haematogenous diffusion through the meningeal branches to the posterior fossa dura from the occipital or ascending pharyngeal arteries. PMID- 12587798 TI - Properly selected patients with multiple brain metastases may benefit from aggressive treatment of their intracranial disease. AB - To determine whether properly selected patients with multiple brain metastases benefit from aggressive treatment of their intracranial disease, we reviewed 52 patients having stereotactic radiosurgery (SRS), tumor resection, or both between April 1997 and March 2000. Tumor histology included lung (n = 18, 35%), breast (n = 11, 21%), renal (n = 6, 12%), melanoma (n = 6, 12%), and other (n = 11, 21%). The median patient age was 58 years, the median Karnofsky performance status (KPS) was 90, and the median number of tumors was three. Twenty patients (39%) had progressed after prior radiation therapy. Treatment included multiple craniotomies and tumor resection (n = 5, 10%), radiosurgery (n = 31, 60%), or resection and radiosurgery (n= 16, 30%). Median survival was 15.5 months. The one and two-year actuarial survivals were 63% and 27%, respectively. Multivariate analysis found radiation therapy oncology group recursive partitioning analysis (RTOG RPA) Class (1 vs. 2/3) correlated with improved survival (Relative risk = 2.60, 95% CI 1.13-5.97, p = 0.03). Class 1 patients (KPS > or = 70, age < 65 years, and controlled primary with no extracranial metastases) survived a median of 19 months whereas Class 3 patients (KPS < 70) survived 8 months. Class 2 patients (all other patients) survived a median of 13 months. Thirty-five patients (67%) had intracranial progression at a median of 8.0 months. Intracranial progression was local (n = 6), distant (n = 23), or local and distant (n = 6); 26 patients with intracranial progression underwent additional brain tumor treatments. Multivariate analysis found patients with radiosensitive tumors (lung, breast, other) had fewer intracranial recurrences compared to patients with radio-resistant tumors (melanoma, renal, sarcoma) (Relative risk = 2.43, 95% CI 1.13-5.10, p = 0.02). The length of survival in our series is quite comparable to historical reports on the management of brain metastasis patients, and supports aggressive intervention for RTOG RPA Class 1 patients and Class 2 patients with controlled primary disease who have a limited number of brain metastases. PMID- 12587799 TI - Effects of 4-aminopyridine on cardiac repolarization, PR interval, and heart rate in patients with spinal cord injury. AB - STUDY OBJECTIVE: To determine the effects of 4-aminopyridine (4-AP) on heart rate and PR, QT, and QTc intervals in patients with longstanding spinal cord injury (SCI). DESIGN: Randomized, active-treatment-controlled, dose level-blinded study, with allocation concealed. SETTING: University-affiliated, tertiary care medical center. PATIENTS: Sixty otherwise healthy male and female outpatients with traumatic SCI of more than 1 year's duration. Intervention. Oral administration and dose titration to tolerance of an immediate-release formulation of 4-AP. MEASUREMENTS AND MAIN RESULTS: The PR interval, heart rate, QT interval, and QTc interval obtained from standard 12-lead electrocardiograms (ECGs) at baseline (before administration of 4-AP) and after 1 month of treatment were compared. The QTc intervals were derived by using Bazett's formula (equation) incorporated into standard computerized analyses of 12-lead ECG printouts. The paired t test was performed to test for the significance of differences between means and variances. No statistically significant differences were noted in heart rate or between ECG time intervals measured at baseline and after 1 month of treatment with 4-AP among all patients with SCI or between subgroups stratified by injury level (tetraplegia, paraplegia) or sex. CONCLUSION: During the 1-month period that 4-AP was administered, the heart rate and PR, QT, and QTc intervals all remained unchanged and stayed well within normal range in comparison to literature-derived control values. 4-Aminopyridine does not appear to influence the length of cardiac time intervals or heart rate and, hence, is unlikely to cause potentially life-threatening ventricular dysrrhythmias when administered long-term and taken orally in dosages of up to 30 mg/day. Specifically, cardiac repolarization (QTc interval) is unaffected in patients with SCI who continuously receive 4-AP for up to 1 month. PMID- 12587800 TI - Reteplase for dysfunctional hemodialysis catheter clearance. AB - STUDY OBJECTIVE: To evaluate the efficacy of reteplase administration in clearing hemodialysis catheters. DESIGN: Open-label, uncontrolled, observational trial. SETTING: University medical center. PATIENTS: Thirty-four patients with end-stage renal disease undergoing long-term hemodialysis. INTERVENTION: Patients had dual lumen, cuffed, tunneled dialysis catheters placed for long-term vascular access. Reteplase 3 U was instilled into each catheter lumen (total dose 6 U) in the first 20 episodes, 2 U in each catheter lumen (total dose 4 U) in the next 20, and 0.5 U in each catheter lumen (total dose 1 U) in the final 45. MEASUREMENTS AND MAIN RESULTS: Over 12 months, 85 episodes of catheter dysfunction were documented. Catheter dysfunction was defined as absence of flow from the catheter lumen, inability to aspirate heparin from the lumen, blood flow rates below 150 ml/minute, or venous pressure greater than 250 mm Hg at blood flow rates below 200 ml/minute. Reteplase was instilled into the catheter lumens and allowed to dwell there until the next hemodialysis session. Successful catheter recanalization was defined as return of aspiration and infusion function allowing dialysis to be completed at blood flow rates above 300 ml/minute. Reteplase restored catheter function in 74 (87%) instances of catheter dysfunction. In the first 40 episodes in which 4- or 6-U doses were given, catheter function was restored in 36 instances (90%). There was no difference in restoration of catheter function between 4 U (18/20, 90%) and 6 U (18/20, 90%). In the last 45 cases in which 1 U was administered, function was restored in 38 catheters (84%). Mean overall dwell times were not different between the first 40 (32 +/- 7 hrs) and the last 45 episodes (33 +/- 10 hrs). The overall mean duration of catheter patency was 45 +/- 39 days. Durations of patency in the three dose groups were not significantly different (44 +/- 38, 46 +/- 40, 45 +/- 39 days). No patient suffered adverse effects related to reteplase. CONCLUSION: Reteplase installation in dysfunctional hemodialysis catheters was effective in restoring catheter function in 87% of episodes. A dose of 1 U appears to be as effective as 4 and 6 U. PMID- 12587801 TI - Heparin resistance as detected with an antifactor Xa assay is not more common in venous thromboembolism than in other thromboembolic conditions. AB - STUDY OBJECTIVE: To identify whether heparin resistance when measured with an antifactor Xa assay is more frequent in patients with venous thromboembolic disease than in patients with other thromboembolic conditions. DESIGN: Prospective observational cohort study. SETTING: A 625-bed, adults-only, private teaching hospital. PATIENTS: Three hundred seventy-two patients receiving intravenous unfractionated heparin for a variety of indications classified as either venous thromboembolism or arterial thromboembolism. INTERVENTION: All patients received an initial unfractionated heparin dosage according to a protocol based on age and estimated blood volume. Targeted antifactor Xa was 0.35 0.65 U/ml. MEASUREMENTS AND MAIN RESULTS: A modest increase in heparin dosage requirement was noted in patients with venous thromboembolism that was entirely related to lower hemoglobin concentration in that group compared with the arterial thromboembolism group. Low hemoglobin concentration was significantly associated with increased heparin resistance regardless of treatment indication. CONCLUSIONS: True heparin resistance seen with an antifactor Xa assay is not more common in venous thromboembolism than in other conditions. Low hemoglobin concentration, rather than treatment indication, appears to be associated with increased heparin resistance. PMID- 12587802 TI - A 24-hour comparison of serum growth hormone concentrations in patients with heart failure versus healthy controls. AB - STUDY OBJECTIVE: To compare endogenous serum growth hormone concentrations over a 24-hour period in patients with chronic heart failure (CHF) and matched controls. DESIGN: Prospective, 24-hour, endogenous concentration comparison. SETTING: Hospital research center. PATIENTS: Eight evaluable patients with nonischemic dilated cardiomyopathy and 10 healthy control subjects, matched for age and sex. INTERVENTION: Over a 24-hour period, blood was drawn from the study participants every 20 minutes for determination of growth hormone. MEASUREMENTS AND MAIN RESULTS: For each patient, the area under the concentration-time curve from time 0-24 hours (AUC0-24), maximum concentration (Cmax), and minimum concentration (Cnadir) of growth hormone were determined. The AUC0-24 and Cmax were 74% (p < 0.05) and 62% (p < 0.05) lower in patients with CHF than in controls, respectively. The Cnadir for all participants was 0 microg/L. Variability in growth hormone concentrations over the 24 hours was considerable for all study participants. CONCLUSIONS: Growth hormone concentrations are suppressed over a 24 hour period in patients with CHF versus healthy controls. Variability in levels throughout the day suggests that a single point evaluation cannot be used to determine deficiency or abundance of growth hormone. PMID- 12587803 TI - Hydromorphone transfer into breast milk after intranasal administration. AB - STUDY OBJECTIVES: To determine the distribution of hydromorphone into breast milk and the potential exposure of the suckling infant, and whether the distribution of hydromorphone into milk can be predicted accurately by a passive diffusion model. DESIGN: Single-dose, pharmacokinetic study. SETTING: University clinical research unit. PATIENTS: Eight lactating, nonsmoking, healthy women aged 24-32 years. INTERVENTION: Hydromorphone HCl 2 mg was given intranasally to the women to characterize its pharmacokinetics and extent of its transfer into breast milk. MEASUREMENTS AND MAIN RESULTS: Plasma and milk samples were analyzed using liquid chromatography with tandem mass spectrometry detection. The milk:plasma ratio (M:P) was calculated as the total area under the concentration-time curve (AUC) of the milk divided by the total AUC of the plasma. Predicted in vitro M:P ratios were calculated using a diffusion model. Protein binding in milk and plasma, partitioning into milk fat (whole milk:skim milk ratios), as well as pH partitioning between plasma and milk were incorporated in the model. Protein binding was determined by equilibrium dialysis. Protein binding was minimal in both milk and plasma, with unbound fractions of 1 and 0.84, respectively There was little partitioning into milk fat, as demonstrated by the whole milk:skim milk ratio of 0.98. The observed and predicted M:P ratios +/- SD for hydromorphone were 2.57 +/- 0.47 and 1.11 +/- 0.28, respectively. The 95% confidence interval for the observed M:P ratio overlapped the confidence interval of the predicted M:P ratio, a finding that supports a role for both passive diffusion and active transport as mechanisms of hydromorphone transfer into milk. CONCLUSION: Hydromorphone distributes rapidly from plasma into breast milk; however, the drug does not partition into fat. The suckling infant would receive approximately 0.67% of the maternal dose of hydromorphone (adjusted for body weight). As this is a limited exposure, further studies are needed to determine any potential impact to an infant who is fed breast milk from a mother treated with hydromorphone. PMID- 12587804 TI - Risk factors for arthralgias or myalgias associated with quinupristin dalfopristin therapy. AB - STUDY OBJECTIVE: To evaluate risk factors for the development of arthralgias or myalgias associated with quinupristin-dalfopristin. DESIGN: Retrospective chart review and case-control analysis. SETTING: An 850-bed tertiary care medical center. PATIENTS: All adult and pediatric patients who had received quinupristin dalfopristin through either a compassionate-use protocol (February 1996-October 1999) or in the year after quinupristin-dalfopristin was added to the hospital formulary (November 1999-October 2000) were included in this study. Case patients were those who developed arthralgias or myalgias while receiving quinupristin dalfopristin therapy; control patients were those who received quinupristin dalfopristin but did not develop arthralgias or myalgias. INTERVENTION: Medical records, pharmacy dispensing information, and microbiology data were reviewed by a physician and a pharmacist, both of whom specialized in infectious diseases. Presence or absence of arthralgias or myalgias was the primary outcome assessed. MEASUREMENTS AND MAIN RESULTS: Quinupristin-dalfopristin was administered to 68 patients during the period defined by the study. Arthralgias and myalgias could not be assessed in 18 of the 68 patients because they were sedated and paralyzed, or they were young children who could not communicate the presence of pain. Univariate analysis demonstrated that significant risk factors for arthralgias or myalgias associated with quinupristin-dalfopristin were female sex, chronic liver disease, receipt of liver transplant, elevated bilirubin level at baseline, major surgery, and receipt of either mycophenolate or cyclosporine. Multivariate analysis demonstrated a strong association with chronic liver disease, receipt of liver transplant, elevated bilirubin level at baseline, and receipt of either cyclosporine or mycophenolate. Of 50 evaluable patients receiving quinupristin dalfopristin, 25 had pain that may have been associated with this antimicrobial agent. CONCLUSION: The mechanism for development of arthralgias or myalgias associated with quinupristin-dalfopristin remains unknown, but these adverse events are more likely to occur in patients with chronic liver disease and those who have received a liver transplant or are receiving cyclosporine or mycophenolate. PMID- 12587805 TI - 2-Methoxyestradiol, a promising anticancer agent. AB - Estrogens occurring naturally in the body are metabolized to catecholestrogens (2 and 4-hydroxyestradiol) by the cytochrome P450 enzymes. 2-Hydroxy catecholestrogens are further metabolized by catechol-O-methyltransferase to 2 methoxyestradiol, which is known to be protective against tumor formation. 2 Methoxyestradiol exhibits potent apoptotic activity against rapidly growing tumor cells. It also possesses antiangiogenic activity through a direct apoptotic effect on endothelial cells. Other molecular mechanisms, including microtubule stabilization by inhibition of the colchicine-binding site, have been reported. The exact mechanism of action of 2-methoxyestradiol is still unclear, but it has been shown to be effective in preventing tumor growth in a variety of cell lines. 2-Methoxyestradiol also possesses cardioprotective activity by inhibiting vascular smooth muscle cell growth in arteries. It has a lower binding affinity for estrogen receptor alpha compared with that of estradiol, and its affinity for estrogen receptor beta is even lower than that of estrogen receptor alpha, thus it has minimal estrogenic activity. 2-Methoxyestradiol is distinct because of its inability to engage estrogen receptors as an agonist, and its unique antiproliferative and apoptotic activities are mediated independently of estrogen receptors alpha and beta. A phase I clinical trial of 2-methoxyestradiol 200, 400, 600, 800, and 1,000 mg/day in 15 patients with breast cancer showed significant reduction in bone pain and analgesic intake in some patients, with no significant adverse effects. Another phase I study of 2-methoxyestradiol 200 1,000 mg/day in combination with docetaxel 35 mg/m2/week for 4-6 weeks performed in 15 patients with advanced refractory metastatic breast cancer showed no serious drug-related adverse effects. A phase II randomized, double-blind trial of 2-methoxyestradiol 400 and 1,200 mg/day in 33 patients with hormone-refractory prostate cancer showed that it was well tolerated and showed prostate specific antigen stabilizations and declines. We have started a phase I clinical trial to explore dosages greater than 1,000 mg/day. PMID- 12587806 TI - Angiotensin II receptor blockers in heart failure. AB - Heart failure remains a significant cause of morbidity and mortality, despite major advances in therapy. Angiotensin II, the principal mediator of the renin angiotensin system, exerts both short-term (e.g., hemodynamic, renal) and long term (e.g., inflammation, cardiac remodeling) effects in the pathophysiology of cardiovascular disease. The effects of angiotensin II appear to be more completely inhibited by angiotensin II receptor blockers (ARBs), which act at the subtype 1 receptor level, than by angiotensin-converting enzyme (ACE) inhibitors because pathways other than that of ACE contribute to the generation of angiotensin II. Evidence demonstrates that ARBs, when added to conventional treatment for patients with heart failure, are associated with a reduction in morbidity and mortality as well as an improvement in quality of life. Clinical trials of ARB therapy indicate that these agents are generally well tolerated, both alone and in combination with other neurohormonal inhibitors. The current role of ARBs in heart failure is as an alternative for patients who cannot tolerate therapy with an ACE inhibitor. A number of ongoing clinical studies are likely to further define or expand the role of ARBs in the treatment of cardiovascular disease. PMID- 12587807 TI - Tiotropium: an inhaled, long-acting anticholinergic drug for chronic obstructive pulmonary disease. AB - Inhaled anticholinergic drugs are considered one of the principal bronchodilator treatments for chronic obstructive pulmonary disease (COPD). Ipratropium bromide is an anticholinergic drug frequently administered for the treatment of COPD. Unfortunately, ipratropium has a short duration of action, requiring administration every 6 hours; this regimen affects adherence to drug therapy. Tiotropium bromide is structurally similar to ipratropium and is under development in the United States. The duration of action of tiotropium is approximately 24 hours, allowing for once-daily dosing. Other than xerostomia being more common with tiotropium than with ipratropium, the safety profiles of these drugs were similar in studied populations. On the basis of its improvements in trough spirometric measurements and improved pharmacokinetic profile compared with that of ipratropium, tiotropium is likely to become the first-line anticholinergic agent in the treatment of patients with COPD. PMID- 12587808 TI - Lack of diagnosis and treatment of osteoporosis in men and women after hip fracture. AB - STUDY OBJECTIVE: To determine whether men and women admitted to a university teaching hospital for a low-trauma hip fracture were diagnosed, evaluated, or treated for osteoporosis during admission or for up to 1 year after admission. DESIGN: Retrospective chart review. SETTING: University of Colorado Hospital, Denver, Colorado. PATIENTS: One hundred eighteen patients admitted with a low trauma hip fracture from January 1993-December 1998. MEASUREMENTS AND MAIN RESULTS: Demographics, medical and social history, prescribed drugs, clinical outcomes, and information regarding the diagnosis, evaluation, and treatment of osteoporosis were abstracted from inpatient medical records for the index hip fracture. Similar data for the first year after the index hip fracture were abstracted from outpatient medical records of patients who had follow-up visits within the hospital system. Mean +/- SD age at the time of fracture was 70 +/- 15 years; 43 patients were men and 75 were women. Eighteen percent of patients had experienced a previous hip fracture, 4% had a history of vertebral fracture, and 6% reported a previous wrist fracture. The diagnosis of osteoporosis was noted in the charts of 14% of the patients at discharge and 26% of patients at follow-up. Only 4% of patients during hospitalization and 9% during follow-up received any evaluative tests for osteoporosis, including bone densitometry. Subsequent fractures occurred in 12.5% of patients. Documented treatment of osteoporosis was uncommon, with approximately 75% of patients receiving no therapy for osteoporosis on discharge or during follow-up. Women were more likely than men to receive a diagnosis of osteoporosis, bone mineral density testing, and osteoporosis drug therapy. CONCLUSION: In patients with hip fractures, osteoporosis is commonly not diagnosed or treated appropriately PMID- 12587809 TI - Pneumococcal vaccination in hospitalized elderly patients: role of the pharmacist. AB - OBJECTIVES: To identify factors associated with receiving the pneumococcal vaccine before admission among hospitalized elderly patients and to determine patient perceptions about the role of the pharmacist in advocating its use. METHODS: One hundred sixty elderly (> or = 65 yrs) patients hospitalized during the first 4 days of each month over a 5-month period were interviewed in person to obtain data regarding their attitudes toward receiving the vaccine, knowledge about it, access to health care, health status, and interaction with a pharmacist. RESULTS: Half of the patients had received the pneumococcal vaccine before hospital admission. Patient awareness of the vaccine and physician recommendation significantly predicted vaccination; health status and access to care did not. Provider communication to patients regarding the benefit of the vaccine was a significant factor leading to awareness about the vaccine, thereby resulting in a higher likelihood of vaccination. Half of the nonvaccinated patients were willing to receive the vaccine according to the pharmacist's recommendation. Although many patients preferred to receive the vaccine at their physician's office, nearly 40% had no preference or even preferred the pharmacy. However, only half of the patients had interacted with a pharmacist in the past, and only 20% were aware that some pharmacists are certified to administer the vaccine. CONCLUSIONS: Pneumococcal vaccine coverage is low among hospitalized elderly patients despite the presence of many comorbid conditions that place them at risk and are indications for the vaccine. The pharmacist as a provider can help increase vaccination coverage by recommending and offering the vaccine. Improvements in the level of pharmacist-patient interaction and promoting patient care services to the public are needed. PMID- 12587811 TI - Pharmacologic management by clinical pharmacists of behavioral and psychological symptoms of dementia in nursing home residents: results from a pilot study. AB - A pharmacist-based consulting service was developed for the pharmacologic management of behavioral and psychological symptoms of dementia (BPSD) in a nursing home setting. Patients were evaluated using the Behavioral Pathology in Alzheimer's Disease (BEHAVE-AD) rating scale, and pharmacotherapy was selected using a structured approach. Eleven patients were evaluated and treated with various psychotropic drugs. The most commonly administered drug was trazodone at a mean dosage of 70 mg/day (range 50-100 mg/day). Nine of the patients demonstrated satisfactory treatment responses as shown by a decreased BEHAVE-AD score of 30% or more (average BEHAVE-AD scores at baseline and 1 month after treatment were 13 +/- 4 and 4 +/- 3, respectively), and no clinical side effects were observed. The service was well received by the facility staff and primary care providers. These preliminary results suggest that pharmacists can play an important role in the pharmacotherapy of BPSD with positive clinical outcomes. PMID- 12587810 TI - Physician-pharmacist comanagement of hypertension: a randomized, comparative trial. AB - OBJECTIVE: To compare the effectiveness of an evidence-based, systematic approach to hypertension care involving comanagement of patients by primary care physicians and clinical pharmacists versus usual care in reducing blood pressure in patients with uncontrolled hypertension. METHODS: Patients in a staff model medical group with uncontrolled hypertension were randomized to either a usual care (UC) or a physician-pharmacist comanagement (PPCM) group. All physicians in the study received both group and individual education and participated in the development of an evidence-based hypertension treatment algorithm. Physicians were then given the names of their patients whose medical records documented elevated blood pressures (defined as systolic > or = 140 mm Hg and/or diastolic > or = 90 mm Hg for patients aged < 65 yrs, and systolic > or = 160 mm Hg and/or diastolic > or = 90 mm Hg for those aged > or = 65 yrs). Patients randomized to the UC group were managed by primary care physicians alone. Those randomized to the PPCM group were comanaged by their primary care physician and a clinical pharmacist, who provided patient education, made treatment recommendations, and provided follow-up. Blood pressure measurements, antihypertensive drugs, and visit costs/patient were obtained from medical records. RESULTS: One hundred ninety-seven patients with uncontrolled hypertension participated in the study. Both PPCM and UC groups experienced significant reductions in blood pressure (systolic -22 and -11 mm Hg, respectively, p < 0.01; diastolic -7 and -8 mm Hg, respectively, p < 0.01). The reduction in systolic blood pressure was greater in the PPCM group after adjusting for differences in baseline blood pressure between the groups (p < 0.01). More patients achieved blood pressure control in the PPCM than in the UC group (60% vs 43%, p = 0.02). Average provider visit costs/patient were higher in the UC than the PPCM group ($195 vs $160, p = 0.02). CONCLUSIONS: An evidence-based, systematic approach using physician-pharmacist comanagement for patients with uncontrolled hypertension resulted in improved blood pressure control and reduced average visit costs/patient. PMID- 12587812 TI - A survey of herbal use in children with attention-deficit-hyperactivity disorder or depression. AB - OBJECTIVE: To examine whether herbal medicines were given to children or adolescents receiving care for attention-deficit-hyperactivity disorder or depression. METHODS: Between October 2000 and July 2001, a 23-item questionnaire was administered in five community mental health centers in Texas. Parents or primary caregivers of children who received a psychiatric assessment were sought for participation. One hundred seventeen caregivers completed a questionnaire. The main outcome measure was primary caregivers' self-report of the use of herbal therapy in their children. RESULTS: The lifetime prevalence of herbal therapy in patients was 20% (23 patients). Eighteen patients (15%) had taken herbal medicines during the past year. Recommendations from a friend or relative resulted in the administration of herbal medicines by 61% of 23 caregivers. Herbal medicines were given most frequently for a behavioral condition, with ginkgo biloba, echinacea, and St. John's wort most prevalent. Almost 83% of caregivers gave herbal medicines alone, whereas 13% gave herbal medicines with prescription drugs. Most caregivers (78%) supervised the administration of herbal therapy in their children; the children's psychiatrists (70%), pediatricians (56%), or pharmacists (74%) typically were not aware of the use. CONCLUSIONS: Most caregivers supervised herbal therapy in their children, without communication with a health professional. A need exists for better communication between health professionals and caregivers regarding the use of herbal therapy. PMID- 12587813 TI - Impact of hospitalization on blood pressure control in Italy: results from the Italian Group of Pharmacoepidemiology in the Elderly (GIFA). AB - STUDY OBJECTIVES: To evaluate whether hospitalization affects blood pressure control in hypertensive patients, and to identify factors associated with attainment of adequate blood pressure control and with aggressive pharmacologic treatment. DESIGN: Retrospective study. SETTING: Eighty-one hospitals throughout Italy. PATIENTS: A total of 3,304 patients (59% women, 41% men) with a diagnosis of hypertension and uncontrolled blood pressure values at hospital admission. MEASUREMENTS AND MAIN RESULTS: Patients' blood pressures were surveyed during study periods from 1988-1997. Controlled blood pressure was defined according to the sixth report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (systolic < 140 mm Hg, diastolic < 90 mm Hg). Aggressive pharmacologic treatment was defined as an increase in the number of antihypertensive drugs that the patient was receiving at hospital discharge compared with the number of drugs on admission. Adequate blood pressure control was achieved in only 767 (23.2%) patients, but the proportion increased in each subsequent survey Age (odds ratio [OR] for 10-year increase 0.82, 95% confidence interval [CI] 0.76-0.88) and male sex (OR 0.79, 95% CI 0.65-0.94) were associated with reduced likelihood of achieving adequate blood pressure control. In contrast, coronary heart disease (OR 1.38, 95% CI 1.14-1.66), year of survey (1995 vs 1988: OR 1.47, 95% CI 1.19-1.82; 1997 vs 1988: OR 1.27, 95% CI 1.02 1.42), length of hospital stay (medium vs lower tertile OR 1.20, 95% CI 0.96 1.49; higher vs lower tertile OR 1.42, 95% CI 1.13-1.77), and increase in number of antihypertensive drugs prescribed (OR 1.21, 95% CI 1.02-1.42 for one drug increase) were associated with improved blood pressure control. In 1753 (53.1%) patients, the number of antihypertensive drugs increased during their hospital stay. Younger age, fewer drugs on admission, lower comorbidity index, diagnosis of chronic heart failure, lengthy hospital stay, and increasing baseline values of systolic and diastolic blood pressure were associated with aggressive pharmacologic treatment. CONCLUSION: Adequate blood pressure control was achieved in only 25% of patients with hypertension despite a trend toward improvement in recent years. Advanced age was one of the main factors associated with less aggressive pharmacologic treatment and with inadequate blood pressure control. PMID- 12587814 TI - Perceptions of pharmacists as providers of immunizations for adult patients. AB - Pharmacists have been involved with patient care at the Family Medicine Center, affiliated with the Medical University of South Carolina, for over 20 years. In 1999, to add to existing clinical services, pharmacists administered immunizations (influenza and pneumonia) to over 400 adult patients during clinic visits in designated patient care rooms. A few months after the immunization period, both health care providers and immunized patients were asked to respond to a survey regarding their opinions of pharmacist-administered immunizations. Response rates were 71% for health care providers and 16% for all immunized patients. Most (90%) of the health care respondents felt comfortable with pharmacists providing immunizations and thought it was appropriate for pharmacists to provide this service. However, 35% of the providers did not agree that pharmacists should provide immunizations in local pharmacies. Most (97%) of the immunized patients felt comfortable with their provider but did not recall that a pharmacist had administered the immunization. In addition, 64% questioned the qualifications of a pharmacist to administer immunizations, and only 43% felt comfortable having a community pharmacist administer a vaccine. By extrapolation of these data, one can determine that patients do not regard pharmacists as qualified providers of immunizations. Further study of patient perception of pharmacists in this role is being conducted. PMID- 12587815 TI - Neuroleptic malignant syndrome in an adolescent receiving olanzapine-lithium combination therapy. AB - A 16-year-old boy developed fever, generalized rigidity, leukocytosis, and increased serum transaminase and creatine kinase levels while receiving treatment with olanzapine and lithium. When both drugs were discontinued, his fever and rigidity subsided and biochemical irregularities spontaneously returned to normal, without any complications. Classic neuroleptic malignant syndrome (NMS) was diagnosed. Concomitant administration of lithium with olanzapine may place patients at risk for NMS. Clinicians need to be aware of this rare but potentially fatal side effect in patients of all ages, and especially in adolescents receiving both drugs. PMID- 12587816 TI - Diminished anticoagulant effects of warfarin with concomitant mercaptopurine therapy. AB - The drug interaction between mercaptopurine and warfarin is documented, but case reports of the existence or magnitude of this interaction are rare. An increased warfarin dosage was required for a patient receiving 12-week cycles of mercaptopurine for acute promyelocytic leukemia. At the start of mercaptopurine therapy, an upward titration of 25% beyond the warfarin maintenance dosage was required to achieve a therapeutic international normalized ratio. When a cycle of mercaptopurine was completed, a sharp reduction in warfarin dosage was required. The mechanism behind this interaction is unclear. Mercaptopurine may inhibit gastrointestinal absorption of warfarin, or it may induce hepatic enzymes that metabolize the anticoagulant. With dramatic changes in warfarin or anticoagulant requirements, practitioners should be aware of potential thromboembolic sequelae or bleeding complications when these agents are prescribed concomitantly. Frequent monitoring and careful dosage titration are warranted during concomitant administration. PMID- 12587817 TI - Erythropoietin-induced hypertensive urgency in a patient with chronic renal insufficiency: case report and review of the literature. AB - Hypertension is a potentially dangerous side effect of erythropoietin treatment; however, extreme elevations in blood pressure are rare. A 75-year-old woman with chronic renal insufficiency was treated with subcutaneous erythropoietin. Three weeks before she started receiving erythropoietin, her hematocrit was 27.2%; after 5 weeks of treatment, it rose to 45.7%. The patient came to the emergency department and was admitted with hypertensive urgency. During her hospital stay she was treated with nitroglycerin and nitroprusside infusions, extended-release nifedipine, a variety of beta-blockers, clonidine, and furosemide. By day 3, her blood pressure was adequately controlled. Her renal insufficiency may have progressed as a result of the hypertensive episode, which probably was related to erythropoietin administration and the resultant rapid increase in her hematocrit. Erythropoietin dosing should be titrated to increase the hematocrit gradually, and blood pressure should be monitored closely to avoid serious side effects such as hypertensive emergencies. PMID- 12587818 TI - Low-dose pulse cyclophosphamide in the treatment of neuropsychiatric lupus. PMID- 12587819 TI - Neuropsychiatric lupus favourable response to low dose i.v. cyclophosphamide and prednisolone (pilot study). AB - Sixty SLE patients with only primary neuropsychiatric manifestations (NP-SLE), (54 female and six male; mean age 44.5) were compared consecutively. Forty-six of the patients (78.3%) were presented with a combination of neuropsychiatric symptoms. Except for the standard immunoserological test, all patients underwent clinical, neurological and psychiatric examination, electrophysiological tests [EEG (electro-encephalography involves recording and analysis of electrical signals generated by the brain), EP (the evoked potentials method involves analysis of a series of electrical signals generated in parts of the nervous system following stimulation of sense organs and peripheral nerves) and EMNG (electro-myo-neography is a method of measurement of electrical activity arising from muscle fibers and peripheral nerves)]. This method presents a valuable tool for assessment of functional state of peripheral nervous systems and muscles)), and neuroimaging (MRI of the brain). Thirty-seven out of 60 patients with NP-SLE (group I) were treated with a low dose of i.v. cyclophosphamide (200-400 mg per month). The average daily oral dose of prednisone in these patients was 20.5 mg. Group II consisted of 23 patients treated with pridnisone (mean dose 20.5 per day) only or with antimalarials. Patients in the first group showed a considerable clinical and electrophysiological improvement of cerebral function, while there was only a slight or no improvement in the second group. The difference between groups was statistically significant. PMID- 12587820 TI - Enhancement of T cell apoptosis correlates with increased serum levels of soluble Fas (CD95/Apo-1) in active lupus. AB - Peripheral T cell apoptosis is upregulated in active SLE, in parallel with high expression of both membrane-bound and soluble (s) Fas. Previous studies postulated that sFas down-regulates apoptosis in vitro through its blockade of the Fas-L of cytotoxic cells. We have investigated the extent of apoptosis and sFas levels in 14 patients with active (group A) and 11 with inactive SLE (group B). Fas was predominantly expressed by CD3+ cells from group A, whose increased serological levels of sFas were linearly correlated with the TUNEL positive cell population, whereas low titers paralleled a mild level of apoptosis in group B. This association was also investigated by measuring the effect of sFas on both cell proliferation and caspase activation. We found that incubation with sFas greatly suppressed proliferation of CD3+ cells, especially in group B, and in control cells from healthy donors whose content of CPP32 active products was significantly increased. We postulate that sFas promotes a pro-apoptogen effect, which would explain the high susceptibility to apoptosis in active lupus, and that the apoptosis program itself includes release of sFas to spread the death signal. PMID- 12587821 TI - Lack of evidence of foetal microchimerism in female Spanish patients with systemic sclerosis. AB - Our objective was to study the presence of microchimerism in a series of 47 female Spanish patients with scleroderma (SSc) and to compare with a control group. Polymerase chain reaction was used to identify Y-chromosome sequences in DNA extracted from peripheral blood cells. Y-chromosome sequences were found in DNA from peripheral blood cells in four out of 47 (8.5%) patients with scleroderma (two limited and two diffuse) and in two out of 40 (5%) healthy women (no statistical differences were found). When we compared SSc patients and healthy controls who had had at least one male child, four out of 29 (13.7%) and two out of 26 (7.6%) had microchimerism respectively (no statistically significant differences were found). Patients with both scleroderma and persistent microchimerism had had a male offspring. Foetal microchimerism does not seem to play a major role in most cases of female Spanish patients with SSc. PMID- 12587822 TI - Polymorphisms of the interleukin-4 gene in chinese patients with systemic lupus erythematosus in Taiwan. AB - We aimed to evaluate the relationship between two polymorphisms of the IL4 gene ( 590T/C and intron 3) and systemic lupus erythematosus in Chinese patients in Taiwan. This study included 91 patients with systemic lupus erythematosus (SLE) and 163 unrelated, age matched healthy controls living in the same area. The typing of -590T/C and intron 3 VNTR (variable number of tandem repeats) polymorphisms were performed by PCR-RFLP and PCR, respectively. Allelic frequencies and carriage rates between SLE patients and controls were compared, and the relationship between allelic frequencies and clinical manifestations of SLE was evaluated. The genotype frequencies of IL-4 intron 3 were found to differ significantly between SLE patients with and without discoid rash (chi-square test, P = 0.03 5). The allelic frequency of intron 3 RP1 was significant different in the patients with discoid rash when compared to patients without this clinical feature (OR = 3.70, 95% CI 2.04-6.72, chi2 test, P = 0.029). The RP1/RP1 homozygous carriage was significantly associated with patients with discoid rash when compared to patients without this clinical feature (OR = 6.04, 95% CI 2.81-12.95, P = 0.01). The allelic frequency of -590T was significant different in the patients with discoid rash when compared to patients without this clinical feature (OR = 3.44, 95% CI 1.88-6.31, chi-square test, P=0.04). The T/T homozygous carriage was significantly associated with patients with discoid rash when compared to patients without this clinical feature (OR = 5.41, 95% CI 2.50-11.68, P = 0.02). We describe a novel association between RPI/RPI and T/T homozygous carriage and patients with discoid rash. The role of the intron 3 polymorphism of the IL4 gene in SLE remains unclear and further substantiation based on larger patient samples is needed. PMID- 12587823 TI - Silent nephritis in systemic lupus erythematosus. AB - Silent lupus nephritis (SLN) was investigated in 42 renal asymptomatic patients and compared with 49 untreated patients with overt lupus nephropathy (OLN). Urinary sediment, quantitative proteinuria, creatinine clearance, antinuclear antibodies (ANA), complement, circulating immune complexes (CIC) and renal biopsies were evaluated in all of the patients. Forty-one out of the 42 (97.6%) patients had SLN according to histopathological findings. Results showed that the mean age, female/male ratio and the clinical activity index (SLEDAI) were similar in both groups (P > 0.05). The prevalence of ANA, anti-ds DNA, anti-ENA autoantibodies and C4 serum levels showed no statistical differences between the two groups (P > 0.05). Conversely, in the OLN group, elevated CIC and diminished CH50 and C3 serum levels were significantly different (P < 0.01). WHO class II was the predominant renal lesion in the group with SLN (P < 0.0001), whereas class IV was in the OLN patients (P < 0.0001). We conclude that, in our series, SLN was highly prevalent in renal asymptomatic patients with otherwise systemic lupus erythematosus. Furthermore, abnormal levels of CIC, CH50 and C3 associated with WHO class II suggest a moderate but ongoing activation of immune-mediated renal injury mechanisms. PMID- 12587824 TI - Anti-dsDNA antibody avidity determination by a simple reliable ELISA method for SLE diagnosis and monitoring. AB - High avidity anti-dsDNA antibodies are more specific for SLE diagnosis, and more closely associated with renal involvement than intermediate or low-affinity anti dsDNA antibodies. ELISA methods are largely used to detect anti-dsDNA, but their high sensitivity is inversely related to specificity because they also detect low avidity antibodies. We developed an ELISA assay based on the law of mass action and the competitive binding of dsDNA in solution and coated to microwells with anti-dsDNA antibodies. A simplified Scatchard plot analysis system was used to measure anti-dsDNA antibody avidity which was expressed as apparent affinity constant (Kaa), and quantified in liters per unit (I/U). We prospectively studied 101 consecutive SLE patients, who were followed for 3 years; three serum samples were sequentially collected from each patient during follow-up for determination of IgG anti-dsDNA antibody concentration, and anti-dsDNA avidity. SLE disease activity was estimated using the European Consensus Lupus Activity Measure (ECLAM) index. Sera from 100 healthy subjects and 133 patients with other connective tissue diseases or infectious diseases were also assayed as controls. The mean Kaa in SLE patients was 65.2 +/- 47.3 l/U, with no variations over time. Anti-dsDNA-positive SLE patients had higher Kaa values (79.1 +/- 46.8) than anti dsDNA negative patients (27.2 +/- 20.1; P < 0.001). No correlation emerged between anti-dsDNA avidity and the ECLAM activity index score. Avidity was significantly higher in patients with renal involvement vs patients without this complication (78.2 +/- 50 vs 59.9 +/- 45.6 l/U; P = 0.0013). This simple ELISA method could be very useful in the diagnostic phase to differentiate high avidity anti-dsDNA autoantibodies that are characteristically found in SLE patients from low avidity antibodies that can also be found in other inflammatory diseases. Moreover, our data confirm the predictive value of high avidity anti-dsDNA antibodies for the development of lupus nephritis. PMID- 12587825 TI - Anti-beta2-glycoprotein I autoantibodies require an antigen density threshold, consistent with divalent binding. AB - Autoantibodies binding beta2-glycoprotein I (B2GPI) are an important finding in the antiphospholipid syndrome. While antibodies from mice or rabbits immunized with B2GPI readily bind B2GPI coated on a polystyrene microwell plate, anti-B2GPI autoantibodies only do so when using a modified microwell plate with a negatively charged surface. This study demonstrates that, for the detection of anti-B2GPI autoantibodies in an ELISA using modified plates, an antigen coating concentration threshold exists, such that minimal or no binding occurs below a certain coating concentration of antigen, even though antigen is easily demonstrable on the plate. This is consistent with the hypothesis that autoantibodies require divalent binding to B2GPI for detection, as sufficient antigen density for two protein molecules to be sufficiently close to enable divalent binding would only be expected to occur at higher coating concentrations. Several mutant forms of B2GPI developed for epitope determination experiments are shown to have decreased binding to microtitre plates compared to wild-type. If wild-type and mutants are assayed for antibody binding near the threshold a significant diminution in binding to mutants occurs that is the result of inadequate binding to the plate, but could be misinterpreted as the result of interruption of an epitope by the mutation. PMID- 12587826 TI - Prevalence of T cell receptor zeta chain deficiency in systemic lupus erythematosus. AB - T cells from patients with systemic lupus erythematosus (SLE) display antigen receptor-mediated signaling aberrations associated with defective T cell receptor (TCR) zeta chain expression. We determined the prevalence of TCR zeta chain deficiency in SLE from a large cohort of unselected racially diverse patients with different levels of clinical disease activity as determined by SLE Disease Activity Index (SLEDAI). Our data show that the occurrence of TCR zeta chain deficiency is 78% in SLE patients. There was no relationship between the deficiency of TCR zeta chain and the SLEDAI scores or theapy. TCR zeta chain deficiency was also not associated with age, race or gender and persisted over a 3 year follow-up period. Thus, there is a high prevalence of TCR zeta chain deficiency in SLE patients that is independent of disease activity, and persists over time indicating an important role for TCR zeta chain deficiency in SLE pathogenesis. PMID- 12587827 TI - Long-term effectiveness of danazol corticosteroids and cytotoxic drugs in the treatment of hematologic manifestations of systemic lupus erythematosus. AB - The purpose of this study was to compare the long-term effectiveness among danazol, corticosteroids, cytotoxics, and dapsone in the treatment of hematological manifestations of systemic lupus erythematosus (SLE). Medical charts of all patients seen at the Rheumatic Disease Unit from January to December of 1998 were reviewed. Patient characteristics, disease and treatment information were collected. The main outcome measures were the cause of and time to discontinuation of drugs used to treat hematological manifestations of SLE resulting from all causes, mainly toxicity and inefficacy or both. Bivariate analysis including one-way ANOVA and chi2 tests were used to compare differences between means and proportions, respectively. Survival curves among the different drugs were evaluated using the Kaplan-Meier method. Multivariate analysis (Cox regression) was used to adjust for potential confounders. After all medical records were reviewed 41 cases were eligible. Two cases had hemolytic anemia, 34 had thrombocytopenia, and five had both. These cases had received a total of 121 cycles of treatment at different times and they represent the study population (corticosteroids n = 37, danazol n = 51, citotoxic drugs n = 29, and dapsone n = 4). Crude rates of discontinuations due to any cause, toxicity and inefficacy werenot statistically significant among the drugs. However, the Kaplan-Meier curves showed statistically significant difference for discontinuations due to all causes as well as inefficacy. Prednisone and cytotoxic drugs had the lowest probability of continuation. In contrast, there were not statistically significant differences among the drugs with respect to first relapse. This is the first study examining the long-term termination rates of several drugs used to treat hematological manifestations of SLE. Using rates of discontinuation adjusted for time there were statistically significant differences among the drugs. Danazol had the highest probability of continuation. PMID- 12587828 TI - Antiphospholipid antibodies and stroke in Down syndrome. AB - Two patients with Down syndrome and the primary antiphospholipid antibody are described. One patient had a vasculopathy similar to that previously described as Moyamoya. Down syndrome is characterized by immune defects including a tendency to autoimmune phenomena. This report extends the scope of these observations and particularly draws out the potential role of antiphospholipid antibodies. Indeed antiphospholipid antibodies may well explain the well-known association of Down syndrome and stroke. PMID- 12587829 TI - Bullous SLE: response to methotrexate and relationship with disease activity. AB - We report on a case of 40-year-old female with systemic lupus erythematosus (SLE) who developed a severe bullous eruption on sun-exposed areas. The bullous manifestation was associated with a flare of lupus serologies, whereas the previous manifestations of the disease were quiescent. Due to prior intolerance to many drugs, she was given oral methotrexate (10 mg/week). The drug administration was followed by a rapid and full resolution of cutaneous lesions. Five months later she developed a class III lupus nephritis. Our case is the first report of efficacy of methotrexate in bullous SLE and rises the question of the relationship between bullous eruption and SLE activity. PMID- 12587830 TI - Primary antiphospholipid antibody syndrome masquerading as progressive supranuclear palsy. AB - We report a case of a patient with clinical features of progressive supranuclear palsy and raised titres of anticardiolipin antibodies in blood, thrombocytopenia and livedo reticularis on skin. Magnetic resonance imaging of brain and isotope scan of brain were distinctive of vascular disorder. PMID- 12587831 TI - Angio-Behcet with severe cardiac involvement. PMID- 12587832 TI - Metalaxyl: persistence, degradation, metabolism, and analytical methods. AB - Metalaxyl is a systemic fungicide used to control plant diseases caused by Oomycete fungi. Its formulations include granules, wettable powders, dusts, and emulsifiable concentrates. Application may be by foliar or soil incorporation, surface spraying (broadcast or band), drenching, and seed treatment. Metalaxyl registered products either contain metalaxyl as the sole active ingredient or are combined with other active ingredients (e.g., captan, mancozeb, copper compounds, carboxin). Due to its broad-spectrum activity, metalaxyl is used world-wide on a variety of fruit and vegetable crops. Its effectiveness results from inhibition of uridine incorporation into RNA and specific inhibition of RNA polymerase-1. Metalaxyl has both curative and systemic properties. Its mammalian toxicity is classified as EPA toxicity class III and it is also relatively non-toxic to most nontarget arthropod and vertebrate species. Adequate analytical methods of TLC, GLC, HPLC, MS, and other techniques are available for identification and determination of metalaxyl residues and its metabolites. Available laboratory and field studies indicate that metalaxyl is stable to hydrolysis under normal environmental pH values, It is also photolytically stable in water and soil when exposed to natural sunlight. Its tolerance to a wide range of pH, light, and temperature leads to its continued use in agriculture. Metalaxyl is photodecomposed in UV light, and photoproducts are formed by rearrangement of the N-acyl group to the aromatic ring, demethoxylation, N-deacylation, and elimination of the methoxycarbonyl group from the molecule. Photosensitizers such as humic acid, TiO2, H2O2, acetone, and riboflavin accelerate its photodecomposition. Information is provided on the fate of metalaxyl in plant, soil, water, and animals. Major metabolic routes include hydrolysis of the methyl ester and methyl ether oxidation of the ring-methyl groups. The latter are precursors of conjugates in plants and animals. In soils the most relevant metabolite is the metalaxyl acid, which is formed predominantly by soil microorganisms. Plant uptake, microbial degradation, photodecomposition, and leaching are the major route of metalaxyl dissipation. It has a tendency to migrate to deeper soil horizons with a potential to contaminate groundwater, particularly in soils with low organic matter and clay content. Therefore, precautions should be taken for the continuous application of metalaxyl to crops. If use of metalaxyl is greately increased, the risk of occurrence in groundwater must be reassessed, as by monitoring studies in the most vulnerable areas in main use regions. The R-isomer of metalaxyl (mefenoxam) has recently been registered as the only active compound. Therefore, quantitative studies on the fate of this specific isomer are needed, including appropriate analytical methods. As the use rates of mefenoxam are approximately one-half those recommended for metalaxyl and mefenoxam dissipates more rapidly, concerns for mefenoxam reaching groundwater are even less justified. PMID- 12587833 TI - Indoor household pesticides: hazardous waste concern or not? AB - Many indoor household pesticides are efficient and useful tools for a variety of functions necessary to maintain clean, sanitary, and pleasant homes and institutional facilities, and to provide significant public health benefits. They do so by incorporating active ingredients and formulation technology that have not been associated with significant environmental impact in use or when disposed in landfills. Chemical and environmental fate properties, toxicological characteristics, and use patterns of indoor household pesticides that distinguish them from other categories of pesticides which have been associated with environmental contamination should be recognized when HHW policy is debated and established by governmental agencies. Most indoor household pesticides as defined here should not be considered hazardous waste or HHW because those relatively few containers, often no longer full, that have been disposed with MSW over the years have not been associated with environmental contamination. The tiny amounts of those product residues that will reach MSW landfills have been shown, in general, not to have chemical or environmental fate characteristics that would make them susceptible to leaching. Those that do have the potential to leach based on these characteristics, in most cases, do not represent a threat to human health based on toxicological considerations. However, compounds such as propoxur, which are very mobile and relatively persistent in soil and in addition have been associated with significant potential health effects, may be targeted by the screening process as described here and could be selected for further investigation as candidates for special waste management status (such as HHW). Our analysis and recommendations have not been extended to the many types of lawn and garden pesticides that are commonly used by homeowners and are frequently brought to HHW programs. However, their potential for groundwater contamination could also be judged using the same technical considerations as applied in this review to indoor household pesticides. In light of the very high costs of diverting wastes from the MSW stream and into HHW programs, it is recommended that, as a matter of public policy, all categories of household waste that might be considered as HHW be carefully and objectively evaluated for their potential to harm public health or the environment after disposal at MSW landfills. PMID- 12587834 TI - Mercury modeling to predict contamination and bioaccumulation in aquatic ecosystems. AB - In response to increasing scientific evidence of the environmental toxicity of mercury and its organic compounds, this study reviews the state of knowledge about the mercury cycle in aquatic systems. It describes the aquatic chemistry of mercury and discusses the importance of biological and physicochemical parameters such as pH, temperature, dissolved organic carbon, oxygen concentration, mercury and methylmercury concentration and availability, as well as sulfate, manganese, and iron concentration in surfaced waters. There is still a paucity of information on environmental dependency of factors that regulate the formation, destruction, and trophic transfer of methylmercury. This lack has led to numerous studies to define the factors that can influence its bioconcentration and bioaccumulation. This review presents some alternative models for mercury cycling, speciation, and partitioning based on the trend toward "site-specific water quality standards," in which chemical speciation is considered on a site-by site basis. PMID- 12587835 TI - Biomarkers in terrestrial invertebrates for ecotoxicological soil risk assessment. AB - This review has served to present the most recent information on a selected series of biomarker studies undertaken on soil invertebrates during two extensive European-funded scientific consortia, BIOPRINT and BIOPRINT-II. The goals were to develop and validate methods for the analysis of markers of stress in a range of soil-dwelling organisms. We have discussed the potential and limitations of the following invertebrate biomarkers for soil risk assessment purposes: heat shock proteins, histological and ultrastructural markers, metallothioneins and metal binding proteins, esterases, lysosomal integrity, and the novel biomarker histidine. The hsp response in soil invertebrates is especially suitable to indicate the effects of exposure to comparatively low concentrations for a range of toxicants and can be regarded as a biomarker of general stress. The application of MTs and other metal-binding proteins as biomarkers for exposure in soil invertebrates has been well described, and new methods are being developed for analyzing MT induction both at the protein and molecular level, and reliable and reproducible methods are now available. (Cd)-MT is well characterized for the springtails and its MT concentration is a useful biomarker for exposure as well as for effect. For snails, (Cd)-MT can accumulate in the midgut gland over extended periods of time and therefore its concentration is a biomarker not only for recent intoxication but also for events of cadmium exposure that snails may have experienced a long time before the measurement took place. Cellular and histological alterations can be regarded as reflecting the "health" state of a cell, which may be a measure for the presence of toxicants. Histopathological work on terrestrial invertebrates, however, is still scarce. Isozymes have been poorly studied in soil invertebrates despite their promising role as potential biomarkers in aquatic organisms. Among the large diversity of isozymes, the most well studied are esterases that are frequently used a biomarkers of exposure to various classes of pesticides. Many other isozymes offer potentials for biomarker research, such as glucosephosphate isomerase and phosphoglucomutase, both enzymes necessary for the glycolytic pathway. The lysosomal system has been identified as a particular target for the toxic effects of xenobiotics, although it has yet a limited application in soil invertebrates. This marker is nonspecific, responding equally sensitively to organic or inorganic contamination; however, if used in combination with an earthworm immnunocompetence assay such as total immunoactivity of the coelomocytes, then it is possible to be more specific as to the likely nature of contamination. Free histidine was positively correlated with increasing copper exposure and total copper burden in earthworms from a semifield study. Histidine may thus act as a biomarker of exposure. The transient responses and confounding factors of biomarkers obscure a proper interpretation of biomarker responses under field conditions. These factors are still very poorly understood and require more study. For risk assessment purposes it is recommended that the aforementioned biomarkers may show promise when included in a suite of biomarkers among different soil invertebrate species. It is recommended that a risk assessment protocol draw upon ranking of biomarker responses on a defined scale. It is also hoped that the problems outlined in this review will aid the direction of future research on soil invertebrate biomarkers. PMID- 12587836 TI - Role of allergy in the therapeutic response of nasal polyps. AB - The objective of this study was to determine whether the allergy factor affects therapeutic response of nasal polyps. A total of 68 patients were enrolled between 1 October 1999 and 1 January 2002 at the Allergy and Rhinology Clinic, Faculty of Medicine, Songklanagarind Hospital. Allergy skin prick test was performed in order to divide patients into a positive skin test group and a negative skin test group. Their medical history was recorded including age, sex, nasal symptoms, concomitant diseases and medications. Patients in both groups were treated over a 6 week period with Budesonide nasal spray. Nasal symptoms, polyp size, nasal and oral expiratory peak flow were evaluated at each visit. Overall assessment of treatment efficacy was evaluated by patients at 3 and 6 weeks after treatment. The mean value of these variables during treatment and a baseline period were compared within and between groups. After 3 and 6 weeks of treatment of nasal polyps with topical Budesonide nasal spray, nasal symptoms, polyp size, nasal and oral expiratory peak flow index and overall response to treatment were improved within both groups. Comparing the two groups, there were greater improvements in the negative skin test group compared to the positive skin test group in all variables. These differences in variable scores between groups showed a tendency to increase overtime after treatment was terminated. The results demonstrate that nasal polyps with positive allergen skin test had less improvement compared to nasal polyps with negative allergen skin test in all nasal signs and symptoms and these differences in improvement showed a tendency to increase over time after treatment. PMID- 12587837 TI - Longitudinal follow-up of lymphocyte subsets during the first year of life. AB - A longitudinal study of lymphocyte subsets during infancy was evaluated by using the flow cytometric immunophenotyping method. Two hundred and thirteen blood samples were obtained from 92 healthy, full-term infants of the following ages: 1 7 days old (n = 43), 3 months old (n = 55), 6 months old (n = 57) and 11 months old (n = 58). The absolute numbers of CD3+ and CD3+/CD4+ T lymphocytes increased from birth to 3 months of age, and remained stable thereafter. The absolute number of CD3+/CD8+ T lymphocytes increased from birth to 11 months of age. The absolute number of CD19+ B lymphocytes and NK cells increased rapidly (3 months) after birth and continued to increase throughout the study period. However, the changes in the relative counts of lymphocyte subsets did not always correspond with the changes in their absolute numbers. These results demonstrate the age related changes in lymphocyte subpopulations and provide reference ranges for lymphocyte subsets during infancy. PMID- 12587838 TI - Measurement of IGF-1, IGFBP-3 and free IGF-1 levels by ELISA in growth hormone (GH) deficient children before and after GH replacement. AB - Serum insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) levels reflect the growth hormone (GH) status. A few percent of IGF-1 circulate in a free form which is believed to represent the IGF biological activity. We retrospectively studied the changes of serum IGF-1, serum IGFBP-3, and plasma free IGF-1 levels in growth hormone deficient (GHD) children before and after treatment with recombinant human growth hormone (rhGH) for a period of 6 months and 1 year. Twenty-one GHD children (16 boys and 5 girls) who had the mean chronological and bone ages of 7.7 +/- 0.7 and 4.8 +/- 0.6 years, respectively, were treated with a mean rhGH dose of 11.66 +/- 0.42 U/m2 body surface area/week. Serum IGF-1 level increased from 162.5 +/- 42.9 ng/ml before treatment to 252.8 +/- 49.5 ng/ml (p = 0.007) and 282.7 +/- 86.9 ng/ml after treatment for 6 months and 1 year, respectively. Plasma free IGF-1 also increased from 0.38 +/- 0.30 ng/ml before treatment to 1.21 +/- 0.30 (p = 0.001) and 1.17 +/- 0.42 ng/ml after 6 months and 1 year of treatment. However, serum IGFBP-3 did not significantly increase after treatment. In addition, the free/total IGF-1 ratio decreased after treatment with rhGH. The height velocities at 6 months and 1 year after treatment were negatively correlated with plasma free IGF-1 before treatment. In conclusion, therefore, plasma free IGF-1 levels could serve as a good predictor of growth hormone responses. Furthermore, their circulating levels would be modified by serum IGF-1 status, and possibly, IGFBP-3 protease activity. PMID- 12587839 TI - Hepatitis C virus nonstructural 3 protein: recombinant NS3 protein of the Thai isolates as an antigen in a diagnostic assay. AB - Nonstructural 3 (NS3) protein of hepatitis C virus (HCV) is one of the antigens commonly used in diagnostic assays for antibody to hepatitis C virus. However, immune response to the NS3 protein from one genotype may not cross-react with that from other genotypes. In the development of an anti-HCV assay, the NS3 genes from genotypes 1 and 3 commonly found in Thailand were amplified and cloned into a bacterial expression system. These recombinant NS3 proteins were immunogenic and reacted with plasma samples of Thai patients infected with various HCV genotypes. Interestingly, the NS3 proteins from the Thai genotypes could react with 3 plasma samples from HCV infected Thai blood donors, which could not bind to the NS3.1 protein in the commercial HCV immunoblot kit using antigen from HCV genotype 1. This finding supports our prior observation that the appropriate HCV antigens used in a diagnostic assay should be derived from the virus genotypes commonly found in that geographical region. PMID- 12587840 TI - Synergistic cytotoxicity and apoptosis induced in human cholangiocarcinoma cell lines by a combined treatment with tumor necrosis factor-alpha (TNF-alpha) and triptolide. AB - Cholangiocarcinoma is known to be relatively resistant to chemotherapy. One alternative approach is to use a combination of an immunomodulating agent with an anticancer drug. Here we studied the synergistic actions of TNF-alpha and triptolide (a diterpene epoxide prepared from Tripterygium wilfordii), previously shown to have antitumor activity against hamster cholangiocarcinoma (CCA) cells. Three human CCA cell lines (HuCCA-1, HubCCA-1, KKU-100 cell lines) were subjected to a combined treatment of TNF-alpha (0.1-10 ng/ml) and triptolide (5-50 ng/ml) for 24 hours in microculture plates. The combination of TNF-alpha and triptolide had a significantly increased cytotoxic activity over that of triptolide alone (p < 0.05). Under the same conditions, TNF-alpha by itself was not cytotoxic to these cell lines. Similarly, the combined treatment could also accelerate apoptotic cell death in all three human cholangiocarcinoma cell lines. The combined treatment of TNF-alpha at 10 ng/ml and triptolide at 50 ng/ml for 6-10 hours achieved a percentage of apoptotic cells shown by DAPI staining of 18-65%, compared to only 6-20% apoptotic cells for triptolide alone. Analyzing the possible mechanisms of the combined treatment, we found by Western blot that at 6 hours, there was a poly (ADP-ribose) polymerase (PARP) cleavage which was not detectable by the treatment of either TNF-alpha or triptolide alone. The cleavage of PARP was inhibited when the cells were pretreated with the enzyme inhibitor AC DEVD-CMK, suggesting that apoptosis induced by the combination of TNF-alpha and triptolide involved activation of caspase 3. These results indicate that apoptosis of human cholangiocarcinoma cell lines as induced by a combination of TNF-alpha and triptolide is mediated through caspase 3 activation. PMID- 12587841 TI - Erythema nodosum as a manifestation of HIV infection. AB - Various musculoskeletal syndromes have been well described in patients infected with the human immunodeficiency virus (HIV). Surprisingly, erythema nodosum, an inflammation of the adipose tissue of the skin, has rarely been described. We report a 24-year-old known case of HIV infection, who developed fever and multiple tender subcutaneous nodules that were proven to be erythema nodosum. The patient also had polymyositis. She responded well to a high dose of corticosteroids. Erythema nodosum should be listed as possible the rheumatic manifestation of HIV infection. PMID- 12587842 TI - Neuropsychiatric manifestations in Thai patients with systemic lupus erythematosus. AB - Neuropsychiatric (NP) manifestations in patients with systemic lupus erythematosus (SLE) [NPSLE] and prognostic factors were studied in 91 patients. There were 98 NP episodes, of which 78 (79.6%) occurred within the first year of the disease. Twenty-six patients (6.7%) had NPSLE as an initial presentation of the disease. There were seizures in 53 episodes (54.1%), psychosis in 13 (13.3%), acute confusion state in 11 (11.2%), abnormal consciousness in 6 (6.1%), transverse myelitis in 6 (6.1%), peripheral neuropathy in 5 (5.1%), cerebral infarction in 2 (2.0%) and aseptic meningitis in 2 (2.0%). Most forms of NPSLE responded well to high dose corticosteroids. Anti-convulsant therapy could be discontinued within a median duration of 3 months after the SLE activity was under control, and without significant recurrence of seizures. The 5-year and 10 year survival rates of patients with NPSLE were 75.9% and 50.6%, respectively. Patients with NPSLE had significantly more cutaneous vasculitis and less arthritis than those without. PMID- 12587843 TI - Paraproteins: a regional South Australian experience. AB - We have performed a systematic review of all new serum and urinary paraproteins detected over a six year period in an immunodiagnostic laboratory serving a population of 400,000 people. Clinical diagnoses and associated laboratory features were ascertained from a computerized laboratory database or from clinical notes. Over the period of study, serum or urine paraproteins were detected in 613 new patients. These consisted of 568 patients with serum paraproteins and 45 patients with urinary monoclonal free light chain (in the absence of a serum paraprotein). These paraproteins occurred more commonly in males and the frequency increased with age. Approximately 30% of the serum paraproteins and 60% of urinary monoclonal free light chain were associated with B cell lymphoproliferative disorders (multiple myeloma, plasmacytoma, Waldenstrom's macroglobulinemia, non-Hodgkins lymphoma, chronic lymphocytic leukemia, etc) with the remainder being labeled as monoclonal gammopathies of uncertain significance (MGUS). At clinical presentation, patients with lymphoproliferative disorders tended to have higher levels of paraprotein, B2 microglobulin, the presence of free urinary light chain and demonstrated molecular size heterogeneity of the paraprotein but there was considerable overlap. A good correlation was noted between paraprotein concentration and viscosity in most patients. In conclusion paraproteins were most frequently encountered in the context of a gammopathy of uncertain significance. Features which suggested lymphoproliferative disorders included higher levels of serum paraprotein (> 15 g/l), elevated levels of B2-microglobulin and the presence of urinary free high chain. However, as much overlap was seen with patients with MGUS, regular monitoring of paraprotein level is considered mandatory in the management of these patients. PMID- 12587844 TI - Anaphylactic shock after traditional Russian beauty-treatment-unpleasant surprise in a strongly penicillin-sensitized patient. AB - Beta-lactam drugs can induce allergic immune responses due to their antigenic determinants, promoting IgE-binding and anaphylactic reactions to penicillin. We report a case of a 44-year-old woman who experienced several severe systemic reactions after being exposed directly or indirectly to penicillin. An anaphylactic shock occurred after anal installation of her daughter's urine, who had been treated with penicillin. Skin testing revealed immediate type reactions to major and minor determinants of penicillin indicating an IgE-mediated sensitization. In vitro tests showed elevated levels of penicillin specific IgE. Clinical features, difficulties in taking history and test options for patients with IgE-mediated sensitizations are briefly reviewed in the context of the presented case of unexpected reactions to penicillin due to an immediate type hypersensitivity. PMID- 12587845 TI - IgA deficiency: a report of three cases from Thailand. AB - Selective IgA deficiency has been reported to be the most common primary immunodeficiency disease in Western countries. A markedly lower frequency of this condition has been reported in the Japanese population. While most of the IgA deficient cases are healthy, some patients develop significant recurrent sinopulmonary infections, allergic disorders and autoimmune diseases. Herein, we report three cases of IgA deficiency among Thai patients, all of whom suffered from chronic sinopulmonary infections. Two of the three patients had absolute IgA deficiency while the third had a partial IgA deficiency. The associated conditions found in these three patients were deficiencies of an IgG subclass, allergic rhinitis and lupus nephritis. The youngest child (5 years old boy with lupus nephritis) expired from Pneumocystis carrinii pneumonia complicated with adult respiratory distress syndrome. PMID- 12587846 TI - Treating mood disorders. PMID- 12587847 TI - Correlates of response to acetylcholinesterase inhibitor therapy in Alzheimer's disease. AB - Cholinesterase inhibitors improve cognition and behaviour in some patients with Alzheimer's disease (AD). Studies that have focused on methods to predict response to anticholinesterase therapy and markers for response are reviewed. Among the possible predictors of improvement in cognitive outcomes are apolipoprotein genotype, pretreatment postural blood pressure drop, quantitative electroencephalography (qEEG) and disease progression rate. Of these, qEEG profile after a single dose of an acetylcholinesterase inhibitor was consistently found to be a good predictor of cognitive response. Studies have assessed baseline behavioural profiles and baseline single-photon emission computed tomographic profiles as possible predictors of improvement of behavioural symptoms of AD, but these require further study. Possible markers of response during drug treatment include red blood cell cholinesterase inhibition, cerebrospinal fluid monoamine measurement, pupillary response and platelet amyloid precursor protein analyses. Although they, too, require further study, the analysis of platelet amyloid precursor protein may have value as a correlate of the putative disease-modifying effects of long-term treatment. Studying correlates of response may help to elucidate the mechanism of action of acetylcholinesterase inhibitors. PMID- 12587848 TI - Dopamine genes and attention-deficit hyperactivity disorder: a review. AB - OBJECTIVE: To review the results of genetic studies investigating dopamine related genes in attention-deficit hyperactivity disorder (ADHD). DATA SOURCES: Papers (association/linkage, meta-analyses and animal model studies) were identified through searches of the PubMed database and systematically reviewed. DATA SYNTHESIS: Consistent results from molecular genetic studies are pointing strongly to the possible link between 2 specific genes, the dopamine transporter (SLC3A6) and the dopamine receptor 4 (DRD4), and ADHD. CONCLUSIONS: The implication of SLC6A3 and DRD4 genes in ADHD appears to be one of the most replicated in psychiatric genetics and strongly suggests the involvement of the brain dopamine systems in the pathogenesis of ADHD. However, more work is required to further these findings by genotype-to-phenotype correlations and identify the functional allelic variants/mutations that are responsible for these associations. The role of other dopamine genes, which may have smaller effects than SLC6A3 and DRD4, needs also to be determined. PMID- 12587849 TI - Effect of loxapine on peripheral dopamine-like and serotonin receptors in patients with schizophrenia. AB - OBJECTIVE: To investigate the effect of loxapine on peripheral dopamine D2-like and serotonin receptor binding and on psychotic symptoms. PATIENTS: Patients (n = 24) meeting the diagnostic criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision, for schizophrenia were selected from an outpatient clinic (age range 18-70 yr). METHODS: Patients were given loxapine (dose determined by a physician) for a period of 12 weeks. There were clinic visits at before treatment began and at 6 weeks and 12 weeks of treatment. Scores on a variety of efficacy and safety scales were recorded at each visit, and blood was drawn for receptor assays. RESULTS: Patients showed significant improvement on most psychiatric assessment scales after 6 and 12 weeks of treatment with loxapine, and both lymphocyte D2-like and 5-HTL2A platelet receptor binding were down-regulated after 6 and 12 weeks. The degree of receptor down-regulation was not significantly correlated with improvements in psychotic symptoms, however. CONCLUSION: Loxapine down-regulated both lymphocyte D2-like and platelet 5-HT2A receptors to the same extent, suggesting that both receptors are involved in the mechanism of action of loxapine in patients with schizophrenia. PMID- 12587852 TI - What treatment options exist for alcohol abuse? PMID- 12587851 TI - Mirtazapine for treatment-resistant depression: a preliminary report. AB - OBJECTIVE: To describe the effectiveness and tolerability of mirtazapine, a noradrenergic and specific serotonergic antidepressant, in the open-label treatment of patients with depression who were resistant to other antidepressant agents. METHODS: The charts of 24 patients who met the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, (DSM-IV) criteria for major depressive disorder and were treated with mirtazapine after partial or nonresponse to standard antidepressants were reviewed for clinical response. Outcome was determined by using the Clinical Global Impressions of Improvement (CGI-I) Scale. RESULTS: Symptomatic improvement was observed in 9 (38%) of 24 patients during an average of 14.1 months of mirtazapine treatment at a mean dose of 36.7 mg/day. Five (21%) patients discontinued mirtazapine because of side effects such as fatigue, weight gain and nausea. Five (21%) patients were receiving combination therapy with another antidepressant when mirtazapine treatment was initiated. CONCLUSIONS: This open-label study suggests that a subgroup of patients with treatment-resistant depression may benefit from mirtazapine treatment. Further controlled studies are required to demonstrate the efficacy of mirtazapine in treatment-resistant depression. PMID- 12587850 TI - Substance use and cognition in early psychosis. AB - OBJECTIVE: To determine the relation between substance use and cognition in individuals experiencing their first episode of psychosis. DESIGN: Prospective cross-sectional and longitudinal study. SETTING: An Early Psychosis Treatment and Prevention Program, an outpatient clinic in a psychiatry department at a university-affiliated hospital. PARTICIPANTS: Individuals with a psychotic illness who were admitted to an Early Psychosis Program; 266 patients were assessed at initial presentation, 159 at 1 year and 90 at 2 years. Most were outpatients. MEASURES: The effects of substance use (alcohol, cannabis, hallucinogens, cocaine, stimulants) on cognition were assessed. Substance use was determined by DSM-IV criteria, and the Case Manager Rating Scale was used to determine the level of substance use. A comprehensive cognitive battery of tests was used, and the Positive and Negative Syndrome Scale for schizophrenia was administered to all subjects to determine levels of positive and negative symptoms. RESULTS: Overall, both cross-sectionally and longitudinally, there were no significant associations between cognitive functioning and the use of various substances. Substance use was associated with higher positive symptoms. CONCLUSIONS: Individuals with psychotic disorders who show mild-to-moderate abuse of substances, in particular alcohol and cannabis, do not exhibit more cognitive impairment than those who do not do use the substances. However, substance use may have other detrimental effects on the process of the psychotic illness. PMID- 12587853 TI - Annotation: The use of psychotropic medications in children: an American view. AB - BACKGROUND: Psychotropic medications have become an integral component in the treatment of children with mental illnesses. METHODS: Selective reviews of the empirical evidence for the efficacy of psychotropic medications and studies of their use patterns were reviewed. RESULTS: Very strong efficacy for at least the short-term benefits and safety of stimulant medications was found and some good efficacy and safety evidence for the treatment of anxiety and depressive disorders with seratonin reuptake inhibitors (SSRI) was also found. Efficacy for tricyclic antidepressants to treat attention deficit hyperactivity disorder was found but the presence of significant side effects makes them less the drugs of choice. Other medications are presented but with less rigorous evidence. Studies of use found that stimulant medications are extensively prescribed in the US by both psychiatrists and primary care physicians. SSRI are also prescribed extensively but not to the extent of stimulants and are more frequently prescribed by psychiatrists. CONCLUSIONS: There is now good evidence for the efficacy of some psychotropic agents and their use is an integral component in the management of childhood mental illnesses. PMID- 12587854 TI - Annotation: The use of psychotropic medications in children: a British view. AB - BACKGROUND: Prescribing practices relating to the use of psychotropic medication with mentally disordered children have changed significantly in Britain over recent years. METHODS: I conducted a review of the modest body of empirical data available relating to the prescribing practices of child psychiatrists, paediatricians and general practitioners (primary care physicians). The data were obtained primarily from postal questionnaire studies but also from British drug studies and a government-sponsored evaluation of the efficacy of stimulant medication. Postgraduate training guidelines for the three principal clinical disciplines are also discussed. RESULTS: Systematic evaluation of prescribing practices has a relatively short history. All the studies reviewed demonstrated consistent methodological weaknesses, the most important of which was reliance upon retrospective reports of prescribing practices from clinicians with no analysis of actual prescription data. No studies relating to the general use of psychotropic medication by paediatricians were found. Child psychiatrists and general practitioners appear to be using a range of drugs for a range of conditions; however, there was evidence of intra- and interdisciplinary variations in practice. It was also evident from the general practitioner data that drug treatments were frequently used for conditions best managed with behavioural methods (e.g., common sleep problems and enuresis). Government prescription data relating to methylphenidate use in ADHD reveal a dramatic rise over the past ten years. Currently, most child psychiatrists use this treatment compared to approximately half the profession only seven years ago. The use of newer antipsychotic agents as well as the SSRI antidepressants appears to be growing in child psychiatric practice. A majority of clinicians surveyed believed that medication was an important treatment modality but also felt that they were relatively unskilled in the field and requested further training. CONCLUSIONS: Overall, a picture of both a growing and better informed use of psychotropic medication is emerging in Britain despite shortcomings in postgraduate training. Future research needs to evaluate prescribing practice in a more objective manner in order to improve training and also service developments in the field. PMID- 12587855 TI - Normal variation and abnormality: an empirical study of the liability distributions underlying depression and delinquency. AB - BACKGROUND: Scale scores in studies of emotional and behavioural problems often possess highly skewed distributions. The long upper tails of these distributions place a small proportion of the population at some distance from the main body of the distribution. This invites an interpretation of their forming an abnormal group, one that may be qualitatively distinct. METHODS: Item-response models were fitted to data on parent and self-rated depression and delinquency from four large samples of children or adolescents. RESULTS: We found that underlying liability distributions show very little or no evidence of non-normality. CONCLUSIONS: The results suggest that (i) the skewed nature of the scale scores may be largely measurement artefacts, (ii) the distributions provide no evidence of a qualitatively distinct process generating abnormality as compared to normal variation and (iii) for characterising the whole distribution, including normality and abnormality, the selection of items in typical current assessments of emotional and behavioural problems is not optimal. PMID- 12587856 TI - Perceptions of stressful life events in Northern Irish school children: a longitudinal study. AB - BACKGROUND: Adults' perceptions of stressful life events have been acknowledged as important moderators of the stress adjustment relationship. Until recently, however, there has been a lack of research on children's perceptions of negative life events. This study assesses children's own perceptions of the stressfulness of negative familial, academic and social events as well as events related to the political conflict in Northern Ireland. METHOD: Developmental changes in children's perceptions of events are traced over time. One hundred and sixty 8 year-old children completed a self-report measure of the perceived stressfulness of a range of negative life events. The sample was drawn from schools in the Greater Belfast area to include children of both genders, primary religious affiliations in Northern Ireland (i.e., Protestant and Roman Catholic) and of varying socio-economic status. Three years later, 113 of these children, then aged 11, were traced through the school system and completed the same measure. RESULTS: Children's perceptions of stressful events are related to a host of social factors. Girls viewed many negative events as more stressful than their male counterparts. Roman Catholic and Protestant children differed in their perceptions of conflict-related events. Perceptions of various types of negative experiences were differentially related to socio-economic status and age. CONCLUSION: Personal, social and situational factors differentially determine children's perceptions of negative life experiences. PMID- 12587857 TI - Reliability of the ICD-10 classification of adverse familial and environmental factors. AB - BACKGROUND: The tenth revision of the International Classification of Diseases (ICD-10) contains a number of categories and guidelines for coding different types of adverse familial and environmental situations. These categories have been selected on the basis of empirical evidence that the adverse situations might represent important psychiatric risk factors. Prior studies, using written case vignettes or video- or audiotaped semi-structured interviews, showed that the interrater reliability of these classifications is satisfactory. METHOD: We have tested the interrater reliability of the categories in a multicentre study by using an 'in vivo' design. Classifications were performed in day-to-day practice, using information that is normally available. Three hundred children from 0 to 13 years were involved in the study. These cases had been admitted to various institutes for children with psychiatric disorders, developmental delays and/or adverse psychosocial circumstances. Two clinicians and a student classified the psychosocial situation of each child. In total, 51 clinicians and 6 undergraduate students were involved in the study. RESULTS: It was found that, with the exception of more or less objective categories, the reliability is not satisfactory. Post hoc analyses showed that the insufficient reliability is only partly due to confounding factors: no clear indications were found for information variance, but observation variance may have played a part. Having additional information from two family questionnaires hardly contributes to a better reliability. CONCLUSIONS: The reliability of the psychosocial axis of the ICD-10 is not satisfactory if tested in day-to-day practice. In comparing this result with other reliability studies, it seems that the absence of adequate information to code the psychosocial axis may be a fundamental problem in obtaining sufficient interrater agreement. Apparently, the classification of this axis requires information that is often not available in common practice. PMID- 12587858 TI - Sensitivity to interference and response contingencies in attention deficit/hyperactivity disorder. AB - BACKGROUND: Current theories on ADHD suggest executive and motivational deficits, but it remains unclear whether these are separate deficits or a unitary deficit. METHOD: ADHD children and matched controls performed on a hybrid disjunctive choice reaction time task in which target stimuli could be surrounded by flankers signaling either the appropriate response, the competing response, or response inhibition. The task was performed under three conditions; reward only, reward and occasional punishment, and equal probability of reward and punishment. Heart rate and skin conductance measures were taken during task performance. RESULTS: Contrary to control children, ADHD children slowed their responses when flankers cueing the appropriate response surrounded the stimulus. Flankers cueing incorrect responses further slowed ADHD children relative to control children. ADHD children also responded less accurately under the threat of punishment. Phasic heart rate did not differ between groups, but immediate reward feedback induced greater heart rate responses in control than in ADHD children. Contrary to expectations, groups did not differ in skin conductance responses. CONCLUSIONS: ADHD children appear deficient in approach tendencies in the presence of imminent reward, rather than unresponsive to punishment or negative feedback. Executive inhibition and motivational inhibition seemed to exert separate effects on behavior of children with ADHD. PMID- 12587859 TI - Parental mental health and children's adjustment: the quality of marital interaction and parenting as mediating factors. AB - BACKGROUND: Research has put emphasis on the process of transmission of mental health problems from parents to children. This study examines the specificity of the interpersonal relationships mediating these symptoms. METHOD: Information about parent and child mental health, marital interaction, and parenting was received from 527 mothers and fathers. Information about child mental health was also received from their 12-year-old children (260 girls and 267 boys). RESULTS: The results confirm that parental mental-health problems can compromise a mother's and father's parenting abilities and represent a threat to their children's adjustment. The results suggest that the different types of parental mental-health problems initiate specific paths between parental and child mental health problems. The results also reveal examples of how the mediation may depend on both the parents' and the children's gender. CONCLUSIONS: The results further suggest that opposite-sex parenting is important to children's adjustment during the years of early adolescence. KEYWORDS: Child development, epidemiology, gender, marital relationships, mental health, parenting. PMID- 12587860 TI - Expressed emotion, attributions and depression in mothers of children with problem behaviour. AB - BACKGROUND: This was an initial study seeking to examine the relationship between Expressed Emotion (EE), spontaneous causal attributions and depression in mothers of children referred for problem behaviour. METHOD: Sixty-one mothers were interviewed using the Camberwell Family Interview (CFI). The CFI was coded for maternal EE and spontaneous causal attributions regarding the child's behaviour. Self-report measures of child problem behaviour and maternal depressive symptoms were also completed. RESULTS: Consistent with previous research, high EE mothers, compared to low EE mothers, were more likely to make attributions thatjudged the cause of problem behaviour to be personal to and controllable by the child and also made more 'child-blaming' attributions than low EE mothers. Mothers' scores on the Beck Depression Inventory were found to be associated with 'child-blaming' attributions and higher levels of EE. Regression analyses did not support the hypothesised role of attributions as a mediator between depression and EE but did identify EE as a potential mediator in the relationship between maternal depressed mood and ratings of child problem behaviour. CONCLUSIONS: These results indicate the relevance of both EE and attributions in mothers of children with problem behaviour and suggest that maternal depressed mood is an important factor which is related to both of these. PMID- 12587861 TI - The pitch of maternal voice: a comparison of mothers suffering from depressed mood and non-depressed mothers reading books to their infants. AB - BACKGROUND: Research suggests that storybook reading promotes language development and that there is a relationship between maternal affective responses in relation to infant affect and language development. The purpose of this study is to relate maternal paralinguistic and verbal behaviour during storybook reading to maternal mood state. METHOD: Mothers (n = 32) reporting depressed mood (as measured by the Edinburgh Postnatal Depression Scale) were matched on age of baby (mean age = 6 months, mean age = 10 months), sex of baby, educational status of mother and parity with 32 non-depressed mothers. They were video- and audio taped in their homes while reading a picture-book to their infants. Maternal textual and extra-textual utterances were transcribed and analysed in terms of mean length utterance (MLU), fundamental frequency and pitch modulation. RESULTS: There was an interaction between psychological well being and age group with regard to MLU for text read. Non-depressed mothers had a smaller MLU for younger babies in comparison with older babies, while depressed mothers showed no difference in their MLU. There was a main effect of psychological well being with depressed mothers speaking with a higher mean pitch and more modulations in their pitch, in comparison with non-depressed mothers. Furthermore, there was a significant interaction of the psychological well being of the mother and the mean fundamental frequency used when reading the text and when speaking to their child during the picture-book session. CONCLUSIONS: These differences in maternal speech indicate that mothers who are depressed are less attuned to their infants which might force the infant into self-regulatory patterns that eventually compromise the child's development. PMID- 12587862 TI - How similar are correlates of different subclinical eating problems and bulimia nervosa? AB - BACKGROUND: This study examined the physical, parental, and psychological correlates of subtypes of high EAT-26 scores and bulimia nervosa (BN) in 155 girls/young women seen at two times over a 6-year period (ages 16 and 22). Participants were from white, upper-middle-class families. METHODS: In young adulthood, participants were interviewed for lifetime history for an eating disorder; participants completed questions assessing subclinical problems and correlates at both times of assessment. Based on their EAT-26 scores and diagnostic status for BN at each time, girls were classified into the following groups: True Positive (BN and high EAT-26 scores), False Positive (high EAT-26 scores and no BN), and True Negative (low EAT-26 scores and no BN). RESULTS: In adolescence, 4% of girls were classified as True Positives BN and 23% were classified as False Positives. Similar distributions were found at young adulthood (i.e., 5% True Positives BN and 21% False Positives). By using cluster analysis, two types of subclinical problems (False Positives) were identified at each time with some girls having only high Eating symptoms and others also having high psychopathology. The 'high Eating symptoms only' cluster was more similar to the True Negative group than was the 'high Psychopathology' cluster at each time. CONCLUSIONS: Individuals in the latter group experienced co-occurrence with depression, resulting in pervasive impairments in psychosocial functioning during both adolescence and young adulthood. PMID- 12587863 TI - Autism and a deficit in broadening the spread of visual attention. AB - BACKGROUND: This study examines if visual attention in autism is spatially overfocused (Townsend & Courchesne, 1994) and if there is an associated deficit in broadening the spatial spread of attention. METHOD: Two crosshairs were presented on each trial separated by a brief (500 ms) interval. There was a modest difference in the lengths of the two hairs in each crosshair and participants had to decide which one was longest. Previous research (Mack & Rock, 1998) has revealed that in making this judgement people spread their visual attention to embrace the whole crosshair. Varying the overall size of each crosshair was intended to control participants' spread of attention. The impact of the size of the first crosshair gave an indication of participants' default setting for the spread of attention. The impact of the size transition between the first and second crosshair gave an indication of the fluency with which participants could change the spatial spread of visual attention. RESULTS: Based on the first proposal it was predicted that individuals with autism (N = 13), relative to ability-matched moderately learning disabled (N = 15) and typically developing individuals (N = 15), would be more accurate and quicker to respond when the first crosshair was small rather than large. However, the results revealed no effects of the size of the first crosshair and no group differences. Based on the second proposal it was predicted that individuals with autism, relative to both control groups, would be less accurate and slower to respond to the second crosshair when the size transition from the first crosshair involved a change from small to large (in comparison with large to large), but would not differ when the change was from large to small (in comparison with small to small). This prediction was confirmed. CONCLUSION: Autism is associated with a deficit in broadening the spatial spread of visual attention. The implications of this for other visual and attentional anomalies observed in autism are discussed. PMID- 12587864 TI - Do children with autism fail to process information in context? AB - BACKGROUND: This research investigated the proposal that children with autism are impaired in processing information in its context. To date, this proposal rests almost exclusively on evidence from verbal tasks. Given evidence of visuo-spatial proficiency in autism in other areas of functioning, it is possible that the ability to use context is spared in the visual domain but impaired in the verbal domain. METHOD: Fifteen children with autism and 16 age and IQ-matched typically developing children were tested on their ability to take account of visual context information (Experiment 1) and verbal context information (Experiment 2) using an adaptation of Palmer's (1975) visual context task. They were also given an adaptation of Tager-Flusberg's (1991) visual and verbal semantic memory task (Experiment 3) and Frith and Snowling's (1983) homograph task (Experiment 4). RESULTS: Experiment 1 showed that children with autism were facilitated by the provision of visual context information. Experiments 2 and 3 showed that the same children were also able to use both verbal context information when identifying words and semantic category information in a verbal task when naming and recalling words. However, in Experiment 4 these children had difficulties with a sentence-processing task when using sentence context to disambiguate homographs. CONCLUSIONS: These findings demonstrate that children with autism do not have a general difficulty in connecting context information and item information as predicted by weak central coherence theory. Instead the results suggest that there is specific difficulty with complex verbal stimuli and in particular with using sentence context to disambiguate meaning. PMID- 12587865 TI - Crop protection science at IACR. PMID- 12587866 TI - Molecular diagnostics for fungal plant pathogens. AB - Accurate identification of fungal phytopathogens is essential for virtually all aspects of plant pathology, from fundamental research on the biology of pathogens to the control of the diseases they cause. Although molecular methods, such as polymerase chain reaction (PCR), are routinely used in the diagnosis of human diseases, they are not yet widely used to detect and identify plant pathogens. Here we review some of the diagnostic tools currently used for fungal plant pathogens and describe some novel applications. Technological advances in PCR based methods, such as real-time PCR, allow fast, accurate detection and quantification of plant pathogens and are now being applied to practical problems. Molecular methods have been used to detect several pathogens simultaneously in wheat, and to study the development of fungicide resistance in wheat pathogens. Information resulting from such work could be used to improve disease control by allowing more rational decisions to be made about the choice and use of fungicides and resistant cultivars. Molecular methods have also been applied to the study of variation in plant pathogen populations, for example detection of different mating types or virulence types. PCR-based methods can provide new tools to monitor the exposure of a crop to pathogen inoculum that are more reliable and faster than conventional methods. This information can be used to improve disease control decision making. The development and application of molecular diagnostic methods in the future is discussed and we expect that new developments will increase the adoption of these new technologies for the diagnosis and study of plant disease. PMID- 12587867 TI - A genomics approach to crop pest and disease research. AB - Genome-wide analyses of gene function and gene expression are beginning to yield valuable information in many areas of biological research, and these genomic tools are now being applied to crop pest and disease research. DNA sequencing of cDNA libraries to generate sets of expressed sequence tags (ESTs) are allowing gene compendiums for crop diseases to be compiled. Annotation of such data collections is also providing a wealth of functional information about gene products through similarities to proteins with known function. The next phase of the functional genomics era will be to employ large-scale techniques to knock out or silence genes in order to synthesize gene-specific mutants for phenotypic analysis and to use micro-array methodology to analyze global gene expression, protein turnover and protein processing during the processes of parasitism and colonization. Application of these technologies promises to accelerate the pace that biological information relevant to crop protection accrues. The ability of researchers to assimilate this information into complex models and workable hypotheses is, thus, set to revolutionize the way we study pests and diseases of crop plants. PMID- 12587868 TI - Manipulation of parasitoids for aphid pest management: progress and prospects. AB - This paper describes research at IACR-Rothamsted on aphid parasitoid responses to semiochemical foraging stimuli, aimed at developing novel ways of manipulating these behaviours to overcome ecological constraints to biological and integrated pest control. Female parasitoids respond both to aphid sex pheromones acting as kairomones, and to aphid-induced plant volatiles, acting as synomones. A range of economically important parasitoid species respond to aphid sex pheromones, and their potential for enhancing parasitization of aphid populations has been demonstrated in the field. Commercial production of the pheromone from the plant Nepeta cataria L has been developed and strategies for its use in arable crops are being investigated. Aphid-induced plant volatiles are released systemically throughout the plant and are aphid species specific, probably induced by elicitors in aphid saliva. Aphid-infested plants can induce uninfested neighbours to release damage-related volatiles, plant-to-plant communication occurring via the rhizosphere. The plant compound cis-jasmone has been identified as a plant signal with potential for aphid control, inducing plant defence mechanisms that both deter colonising aphids and attract parasitoids and predators. Such compounds may represent a new generation of crop protectants and their further investigation and development will be aided by the tools generated by genomic and post-genomic biology. PMID- 12587869 TI - Laboratory evaluation of temperature effects on the germination and growth of entomopathogenic fungi and on their pathogenicity to two aphid species. AB - As part of an approach to select potential mycoinsecticides for aphid biocontrol, we investigated the effects of temperature on the growth, germination and pathogenicity of some hyphomycete fungi. Commercially available mycoinsecticides (based on Beauveria bassiana (Balsamo) Vuillemin and Verticillium lecanii (Zimmermann) Viegas) and other isolates of B bassiana, V lecanii, Metarhizium anisopliae (Metschnikoff) Sorokin and Paecilomyces fumosoroseus (Wize) Brown & Smith were evaluated. The rate of in vitro conidial germination of all isolates was slower at 10 and 15 degrees C than at 20 and 25 degrees C. Similarly, in vitro growth of most isolates was adversely affected at 10 and 15 degrees C. The greatest reduction at 10 degrees C in rates of conidial germination and colony growth, compared with other temperatures, was for M anisopliae isolates. Germination of V lecanii (isolate HRI 1.72) was fastest at 10 degrees C compared with the other fungi. It was also the most pathogenic of three isolates tested against Aphis fabae Scopoli and Myzus persicae Sulzer at 10, 18 and 23 degrees C. Generally, A fabae was more susceptible than M persicae to infection by the fungal isolates tested. A significant interaction between aphid species and temperature indicated that the pathogenic nature of an isolate was dependent not only on the target aphid species but also the temperature conditions of the bioassay. The series of studies, detailed above, allowed a temperature profile to be formed for the different isolates. Verticillium lecanii isolate HRI 1.72 (commercialised as Vertalec) was the most promising isolate selected from results of the series of experiments. Temperature profiles in conjunction with infectivity assays can be useful in selecting appropriate isolates for a particular thermal environment. PMID- 12587870 TI - Variation in response to neonicotinoid insecticides in peach-potato aphids, Myzus persicae (Hemiptera: Aphididae). AB - Laboratory bioassays applying the neonicotinoid insecticides imidacloprid, acetamiprid and nitenpyram against clones of the peach-potato aphid Myzus persicae (Sulzer) demonstrated that these compounds effectively circumvent the known carboxylesterase, modified acetylcholinesterase (MACE) and knock-down (kdr) insecticide resistance mechanisms in this species. However, some clones showed cross-tolerance (up to 18-fold) of these compounds relative to susceptible standards. A survey assessing the frequency of neonicotinoid tolerance in M persicae in the UK, based on samples collected from the field and glasshouses between 1997 and 2000, showed that such tolerance is still rare. Experiments on neonicotinoid-susceptible and -tolerant populations of M persicae under simulated field conditions in the laboratory showed that, although the latter were well controlled by imidacloprid applied at recommended application rates, they were more likely to survive and reproduce when this compound was applied at lower concentrations. Such conditions are probably periodically present in imidacloprid treated field and glasshouse crops. Selection favouring tolerant forms of M persicae could lead to increases in their frequency and the evolution of more potent resistance to neonicotinoids. PMID- 12587871 TI - Mode of action and pesticidal activity of the natural product dunnione and of some analogues. AB - This paper reports the investigation of the insecticidal and fungicidal activity of dunnione, a natural product obtained inadvertently as a by-product of a synthesis programme. Dunnione exhibits no insecticidal activity but has an unusually broad spectrum of antifungal activity. In vitro and in vivo (preventative) activities were comparable to those of several long-established fungicides (eg carbendazim). However, in whole-plant assays, its eradicant activity was unexpectedly low, probably due to poor dose-transfer from leaf surface to fungus. The level of residual activity appears to be influenced by the formulation. Finally, its potential as a lead structure was assessed, and several analogues synthesised which exhibited high activity in the in vitro assays. Mode of-action studies revealed that dunnione exerts its action primarily through initiation of redox cycling. This contrasts to the activity of BTG 505, the biochemical/chemical precursor, which does not initiate redox cycling but instead exhibits both insecticidal and fungicidal activity by inhibiting mitochondrial Complex III. PMID- 12587872 TI - Development of a new management strategy for the control of root-knot nematodes (Meloidogyne spp) in organic vegetable production. AB - The nematophagous fungus, Pochonia chlamydosporia (Goddard) Zare & Gams, has been investigated as a potential biological control agent for use in integrated pest management strategies for Meloidogyne incognita (Kof & White) Chitwood in vegetable crops. The release of the fungus as a biological control agent requires a diagnostic method to monitor its spread in the environment and to gain knowledge of its ecology. Only molecular methods are sufficiently discriminating to enable the detection of specific isolates of fungi in soil. A method to extract DNA from soil was developed to increase the efficacy of PCR-based diagnostic tests that use specific primers. A selected isolate of P chlamydosporia var catenulata was applied at densities similar to those that occur naturally in nematode-suppressive soils. The fungus significantly reduced nematode infestations in soil following a tomato crop, in a strategy that combined the use of the fungus with crop rotation. The survival of the fungus in soil was also examined in controlled conditions in which it remained in soil in densities significantly greater than its original application rate for at least 5 months. Hence, it seems that populations of this fungus may be built up in soil and have significant effects on the regulation of root-knot nematode populations. PMID- 12587873 TI - Characterisation of target-site resistance to ACCase-inhibiting herbicides in the weed Alopecurus myosuroides (black-grass). AB - Resistance to aryloxyphenoxypropionate (AOPP), cyclohexanedione (CHD) and phenylurea herbicides was determined in UK populations of Alopecurus myosuroides Huds. Two populations (Oxford AA1, Notts. A1) were highly resistant (Resistance indices 13-->1000) to the AOPP and CHD herbicides fenoxaprop, diclofop, fluazifop P and sethoxydim, but only marginally resistant to the phenylurea, chlorotoluron. Analyses of acetyl coenzyme A carboxylase (ACCase) activity showed that an insensitive ACCase conferred resistance to all the AOPP/CHD herbicides investigated. Another population, Oxford S1, showed no resistance to sethoxydim at the population level, but contained a small proportion of plants (<10%) with an insensitive ACCase. Genetic studies on the Notts A1 and Oxford S1 populations demonstrated that target site resistance conferred by an insensitive ACCase is monogenic, nuclearly inherited with the resistant allele showing complete dominance. Investigations of the molecular basis of resistance in the Notts A1 population showed that sethoxydim resistance in A myosuroides was associated with the substitution of an isoleucine in susceptible with a leucine in resistant plants, which has also been found in three other resistant grass-weed species (Setaria viridis (L) Beauv, Avena fatua L, Lolium rigidum Gaud). PMID- 12587874 TI - Co-induction of glutathione-S-transferases and multidrug resistance associated protein by xenobiotics in wheat. AB - Herbicide safeners are known to protect monocotyledonous crops from herbicide injury by accelerating the metabolism of herbicides. We have investigated the effects of the safener cloquintocetmexyl, which protects small-grain cereals against the graminicidal herbicide, clodinafop-propargyl. Subtractive suppression hybridisation was used to identify wheat genes which are up-regulated by treatment not only with cloquintocet-mexyl but also with phenobarbital, which is known to stimulate xenobiotic metabolism in animals and plants. DNA sequences of five glutathione transferases (GSTs) belonging to three different classes and a multidrug resistance associated protein (MRP) homologue were identified in the screen. The chemical inducibility of these clones was confirmed by Northern analysis. The MRP protein was shown to be induced by treatments with cloquintocet mexyl and phenobarbital and to be localised to the tonoplast. Since clodinafop propargyl is not known to be metabolised by glutathionylation, the significance of GST induction is interpreted in terms of a generalised response to chemical stress, particularly the generation of active oxygen species. This work establishes herbicide safeners as useful tools for the identification of genes encoding herbicide-metabolising enzymes. PMID- 12587875 TI - Variation in the effects of take-all disease on grain yield and quality of winter cereals in field experiments. AB - Relationships between take-all intensity and grain yield and quality were determined in field experiments on cereal crops using regression analyses, usually based on single-point disease assessments made during anthesis or grain filling. Different amounts of take-all were achieved by different methods of applying inoculum artificially (to wheat only) or by using different cropping sequences (in wheat, triticale or barley) or sowing dates (wheat only) in crops with natural inoculum. Regressions of yield or thousand-grain weight on take-all intensity during grain filling were similar to those on accumulated disease (area under the disease progress curve) when these were compared in one of the wheat experiments. Regressions of yield on take-all intensity were more often significant in wheat than in the less susceptible crops, triticale and barley, even when a wide range of disease intensities was present in the latter crops. The regressions usually had most significance when there were plots in the severe disease category. Thousand-grain weight and hectolitre weight usually responded similarly to total grain yield. Decreased yield was often accompanied by a significant increase in the percentage of small grains. When severe take-all was present in wheat, regressions showed that nitrogen uptake was usually impaired. This was sometimes accompanied, however, by increased percentage nitrogen in the grain as a consequence of smaller grain size with decreased endosperm. Significant effects of take-all, both positive and negative, on Hagberg falling number in wheat sometimes occurred. Significant regressions of yield on take-all assessed earlier than usual, ie during booting rather than grain-filling in wheat and triticale and during anthesis/grain-filling rather than ripening in barley, had steeper slopes. This is consistent with observations that severe disease that develops early can be particularly damaging, whilst the crops, especially barley, can later express tolerance by producing additional, healthy roots. The regression parameters, including maximum potential yield (y-axis intercept) and the extrapolated maximum yield loss, also varied according to the different growing conditions, including experimental treatments and other husbandry operations. These differences must be considered when assessing the economic potential of a control measure such as fungicidal seed treatment. PMID- 12587876 TI - Identification and exploitation of novel disease resistance genes in sugar beet. AB - Sugar beet is a valuable arable crop providing a large proportion of world sugar supplies. It is susceptible to a range of fungal and viral diseases, to most of which there is inadequate genetic resistance. Control instead relies largely on the use of pesticides. To increase the sustainability of sugar-beet cultivation, strong and durable genetic resistance is urgently needed. This review describes an IACR-Broom's Barn programme devoted to evaluating Beta germplasm for novel resistance to major viral and fungal diseases, and its progress in elucidating the numbers and locations of the genes responsible. PMID- 12587877 TI - Herbicide risk assessment for non-target aquatic plants: sulfosulfuron--a case study. AB - Herbicides entering the aquatic environment by spray drift, run-off and leaching to field drains may cause adverse effects on non-target aquatic vegetation. The potential for such effects has typically been evaluated from tests with floating, monocotyledonous Lemna sp. However, concern has been expressed as to whether this species could be used to indicate potential effects on other vegetation types, particularly rooted, submerged, emergent or dicotyledonous species. In 1997, the Centre for Aquatic Plant Management undertook development of new tests based on the additional species, Glyceria maxima (Hartm) Holmb, Lagarosiphon major (Ridl) Moss and Myriophyllum spicatum L. The resulting methodology was used to assess the effects of the sulfonylurea herbicide, sulfosulfuron on these species. Data presented here demonstrate that exposure to initial sulfosulfuron concentrations of 3.33 microg litre(-1) for up to 21 days was tolerated by these species and that adverse effects were observed only when plants were exposed to initial concentrations of 3.33 and 10 microg litre(-1) for 70 days. As the occurrence of such high initial concentrations for long periods is unlikely in the aquatic environment, sulfosulfuron is not expected to have adverse effects on the growth of these species. This study has also demonstrated that G maxima, L major and M spicatum grown in small outdoor tanks can be used successfully to assess the effects of crop-protection products on non-target aquatic flora. PMID- 12587879 TI - A revised guide to abbreviations in peptide science and a plea for conformity. PMID- 12587878 TI - The influence of lipophilicity and formulation on the distribution of pesticides in laboratory-scale sediment/water systems. AB - Pesticide reaching surface waters will be sorbed by sediment. This sorption process and the influence of pesticide formulation have been examined at 10 degrees C in small-scale systems having 2-cm depth of sediment and 8-cm overlying water stirred gently. Eight pesticides (triasulfuron, isoproturon, chlorotoluron, phenmedipham, difenoconazole, chlorpyrifos, pendimethalin and permethrin), spanning a range of physicochemical properties, were applied individually to the water. Sorption equilibrium was reached at between 15 and 30 days, the proportion of pesticide then in the sediment ranging from 20% for the acidic and therefore polar triasulfuron to 97% for the lipophilic permethrin; this behaviour was not influenced by formulation. Sorption coefficients measured in batch tests over 2 h gave good estimates of the equilibrium distribution. Some degradation was observed for all compounds over 90 days; for some compounds and formulations, enhanced degradation occurred after 20 to 60 days. It is concluded that lipophilicity is the chief determinant of pesticide distribution in sediment/water systems. PMID- 12587880 TI - Diketopiperazines in peptide and combinatorial chemistry. AB - Diketopiperazines (DKPs), the smallest cyclic peptides, represent an important class of biologically active natural products and their research has been fundamental to many aspects of peptide chemistry. The advent of combinatorial chemistry has revived interest in DKPs for two reasons: firstly, they are simple heterocyclic scaffolds in which diversity can be introduced and stereochemically controlled at up to four positions; secondly, they can be prepared from readily available alpha-amino acids using very robust chemistry. Here synthetic methods, conformation, as well as applications of DKPs are summarized and discussed critically. PMID- 12587881 TI - The aspartimide problem in Fmoc-based SPPS. Part I. AB - A variety of Asp beta-carboxy protecting groups, Hmb backbone protection and a range of Fmoc cleavage protocols have been employed in syntheses of the model hexapeptide H-VKDGYI-OH to investigate the aspartimide problem in more detail. The extent of formation of aspartimide and aspartimide-related by-products was determined by RP-HPLC. This study included three new Fmoc-Asp-OH derivatives: the beta-(4-pyridyl-diphenylmethyl) and beta-(9-phenyl-fluoren-9-yl) esters and also the orthoester Fmoc-beta-(4-methyl-2,6,7-trioxabicyclo[2.2.2]-oct-1-yl)-alanine. 3-Methylpent-3-yl protection of the Asp side chain resulted in significant improvements with respect to aspartimide formation. Complete suppression was achieved using the combination OtBu side chain protection and Hmb backbone protection for the preceding Gly residue. PMID- 12587882 TI - HRMAS NMR observation of beta-sheet secondary structure on a water swollen solid support. AB - In this paper HRMAS NMR was used to investigate whether peptides on a peptidyl resin swollen in aqueous solution can adopt an intramolecular beta-sheet structure. A model peptide YQNPDGSQA, that was previously shown to adopt such a secondary structure in solution, (Blanco et al, J. Am. Chem. Soc., 1993) was grafted onto three different solid supports that swell in aqueous solution to examine the influence of the resin on the structure. Both parameters of resin loading and pH inside the swollen peptidyl resin proved to be important for the physicochemical behaviour of the peptide on the support. PMID- 12587883 TI - Role of a two-residue spacer in an alpha,beta-Didehydrophenylalanine containing hexapeptide: crystal and solution structure of Boc-val-deltaPhe-Leu-Ala-deltaPhe Ala-OMe. AB - The peptide Boc-Val1-deltaPhe2-Leu3-Ala4-deltaPhe5-Ala6-OMe has been examined for the structural consequence of placing a two-residue segment between the deltaPhe residues. The peptide is stabilized by four consecutive beta-turns. The overall conformation of the molecule is a right-handed 3(10)-helix, with average (phi, psi) values (-67.7 degrees, -22.7 degrees), unwound at the C-terminus. The 1H NMR results also suggest that the peptide maintains its 3(10)-helical structure in solution as observed in the crystal state. The crystal structure is stabilized through head-to-tail hydrogen bonds and a repertoire of aromatic interactions laterally directed between adjacent helices, which are antiparallel to each other. The aromatic ring of deltaPhe5 forms the hub of multicentred interactions, namely as a donor in aromatic C-H...pi and aromatic C-H...O=C interactions and as an acceptor in a CH3...pi interaction. The present structure uniquely illustrates the unusual capability of a deltaPhe ring to host such concerted interactions and suggests its exploitation in introducing long-range interactions in the folding of supersecondary structures. PMID- 12587884 TI - Conformational investigation of alpha, beta-dehydropeptides. XI. Molecular and crystal structure of Ac-(Z)-deltaPhe-NMe2 as compared to those of related molecules. AB - A series of three homologous dimethyldiamides Ac-(Z)-deltaPhe-NMe2, Ac-L-Phe-NMe2 and Ac-DL-Phe-NMe2 have been synthesized and their structures determined from single-crystal X-ray diffraction data. To learn more about the conformational preferences of the compounds studied, the fully relaxed phi, psi conformational energy maps on the free molecules of Ac-deltaAla-NMe2 and Ac-(Z)-deltaPhe-NMe2 were obtained with the HF/3-21G method and the calculated minima re-optimized with the DFT/B3LYP/6-31G** method. The crystal state results have been compared with the literature data. The studied dimethyldiamide Ac-deltaXaa-NMe2 combines the double bond in positions alpha, beta and the C-terminal tertiary amide within one molecule. As the representative probe with deltaXaa = deltaAla, (Z)-deltaLeu and (Z)-deltaPhe shows, in the solid state they adopt the conservative conformation with phi, psi approximately -45 degrees, approximately 130 degrees and with a non-planar tertiary amide bond, whatever the packing forces are. This conformation is located on the Ramachandran map in region H/F, which is of high energy for common amino acids, but not so readily accessible to them. The free molecule calculations on Ac-deltaAla-NMe2 and Ac-(Z)-deltaPhe-NMe2 reveal that, in spite of dissimilar overall conformational profiles of these molecules, this structure is one of their low-energy conformers and for Ac-(Z)-deltaPhe-NMe2 it constitutes the global minimum. So, the theoretical results corroborate those experimental results proving that this structure is robust enough to avoid conformational distortion due to packing forces. In contrast to Ac-deltaXaa-NMe2, the saturated Ac-L/DL-Xaa-NMe2 shows the constancy of the associative patterns but do not prefer any molecular structure in the solid state. PMID- 12587885 TI - More on ethics committees. PMID- 12587886 TI - Energy cost of different skating techniques in cross-country skiing. AB - The aims of this study were to compare the aerobic energy cost of four 'on-snow' skating techniques in cross-country skiing and to examine the relationships between performance and aerobic energy cost. Twelve male skiers from recreational to national standard performed four level skating trials of 6 min duration in random order, each at the same submaximal velocity but with a different skating technique: (1) 'offset' (V1), using a double asymmetrical and asynchronous pole plant as weight is transferred to one ski; (2) 'two-skate' (V2A), where the pole plant is symmetrical; (3) 'one-skate' (V2), where there is a pole plant as weight is transferred to each ski; and (4) 'conventional', without poles. Oxygen uptake (VO2), pulmonary ventilation, the respiratory exchange ratio and heart rate were measured using a K4(b2) portable gas analyser. The aerobic energy cost (VO2/mean speed) and heart rate were higher (P < 0.05) in the one-skate than in the offset condition. This may be explained by the greater and more efficient use of the upper body and the lower variation in centre of gravity velocity in the offset condition. The aerobic energy cost was 5-9% higher (P < 0.01) in the conventional than in the other techniques, probably because of the shorter duration of propulsive forces within a cycle in the conventional skating condition. Moreover, in ski skating, the mechanical efficiency (propulsive forces/total forces) was shown to be higher in the upper than in the lower limbs. The correlation coefficient between performance and aerobic energy cost was significant in the two-skate (r = 0.68, P = 0.02), one-skate (r = 0.72, P = 0.01) and conventional (r = 0.62, P = 0.04) conditions, but not in the offset condition (r = 0.50, P = 0.10). Our results stress the importance of the upper body component in cross country skiing and that the aerobic energy cost discriminates between skiers of different standards. PMID- 12587887 TI - The effect of an intermittent, high-intensity warm-up on supramaximal kayak ergometer performance. AB - It has previously been shown that the metabolic acidaemia induced by a continuous warm-up at the 'lactate threshold' is associated with a reduced accumulated oxygen deficit and decreased supramaximal performance. The aim of this study was to determine if an intermittent, high-intensity warm-up could increase oxygen uptake (VO2) without reducing the accumulated oxygen deficit, and thus improve supramaximal performance. Seven male 500 m kayak paddlers, who had represented their state, volunteered for this study. Each performed a graded exercise test to determine VO2max and threshold parameters. On subsequent days and in a random, counterbalanced order, the participants then performed a continuous or intermittent, high-intensity warm-up followed by a 2 min, all-out kayak ergometer test. The continuous warm-up consisted of 15 min of exercise at approximately 65% VO2max. The intermittent, high-intensity warm-up was similar, except that the last 5 min was replaced with five 10 s sprints at 200% VO2max, separated by 50 s of recovery at approximately 55% VO2max. Significantly greater (P < 0.05) peak power (intermittent vs continuous: 629 +/- 199 vs 601 +/- 204 W) and average power (intermittent vs continuous: 328 +/- 39.0 vs 321 +/- 42.4 W) were recorded after the intermittent warm-up. There was no significant difference between conditions for peak VO2, total VO2 or the accumulated oxygen deficit. The results of this study indicate that 2 min all-out kayak ergometer performance is significantly better after an intermittent rather than a continuous warm-up. PMID- 12587888 TI - Prediction of distance in hammer throwing. AB - The aim of this study was to determine how much the predicted distance of a hammer throw is affected by (1) ignoring air resistance and (2) assuming that the centre of mass of the hammer coincides with the centre of the ball. Three dimensional data from actual throws (men: 72.82 +/- 7.43 m; women: 67.78 +/- 4.02 m) were used to calculate the kinematic conditions of the hammer at release. A mathematical model of the hammer was then used to simulate the three-dimensional airborne motion of the hammer and to predict the distance of the throw. The distance predicted for vacuum conditions and using the ball centre to represent the hammer centre of mass was 4.30 +/- 2.64 m longer than the official distance of the throw for the men and 8.82 +/- 3.20 m longer for the women. Predictions using the true centre of mass of the hammer reduced the discrepancy to 2.39 +/- 2.58 m for the men and 5.28 +/- 2.88 m for the women. Predictions using air resistance and the true centre of mass of the hammer further reduced the discrepancy to -0.46 +/- 2.63 m for the men and 1.16 +/- 2.31 m for the women. Approximately half the loss of distance produced by air resistance was due to forces made on the ball and the remainder to forces made on the cable and handle. Equations were derived for calculation of the effects of air resistance and of the assumption that the centre of mass of the hammer coincides with the centre of the ball, on the distance of the throw. PMID- 12587889 TI - The effects of prior incremental cycle exercise on the physiological responses during incremental running to exhaustion: relevance for sprint triathlon performance. AB - It is common for the physiological working capacity of a triathlete when cycling and running to be assessed on two separate days. The aim of this study was to establish whether an incremental running test to exhaustion has a negative effect after a 5 h recovery from an incremental cycling test. Eight moderately trained triathletes (age, 26.2 +/- 3.4 years; body mass, 67.3 +/- 9.1 kg; VO2max when cycling, 59 +/- 13 ml x kg x min(-1); mean +/- s) completed an incremental running test 5 h after an incremental cycling test (fatigue) as well as an incremental running test without previous activity (control). Maximum running speed, maximal oxygen uptake (VO2max) and the lactate threshold were determined for each incremental running test and correlated with the average speed during a 5 km run, which was performed immediately after a 20 km cycling time-trial, as in a sprint triathlon. There were no significant differences in maximum running speed, VO2max or the lactate threshold in either incremental running test (control or fatigue). Furthermore, good agreement was found for each physiological variable in both the control and fatigue tests. For the fatigue test, there were significant correlations between the average speed during a 5 km run and both VO2max expressed in absolute terms (r = 0.83) and the lactate threshold (r = 0.88). However, maximum running speed correlated most strongly with the average speed during a 5 km run (r = 0.96). The results of this study indicate that, under controlled conditions, an incremental running test can be performed successfully 5 h after an incremental cycling test to exhaustion. Also, the maximum running speed achieved during an incremental running test is the variable that correlates most strongly with the average running speed during a 5 km run after a 20 km cycling time-trial in well-trained triathletes. PMID- 12587890 TI - Sodium bicarbonate ingestion does not alter the slow component of oxygen uptake kinetics in professional cyclists. AB - We examined the effects of pre-exercise sodium bicarbonate (NaHCO3) ingestion on the slow component of oxygen uptake (VO2) kinetics in seven professional road cyclists during intense exercise. One hour after ingesting either a placebo or NaHCO3 (0.3 g x kg body mass(-1)), each cyclist (age, 25 +/- 2 years; VO2max, 74.7 +/- 5.9 ml x kg(-1) x min(-1); mean +/- s) performed two bouts of 6 min duration at an intensity of 90% VO2max interspersed by 8 min of active recovery. Gas exchange and blood data (pH, blood lactate concentration and [HCO3-]) were collected during the tests. In both bouts, the slow component of VO2 was defined as the difference between end-exercise VO2 and the VO2 at the end of the third minute. No significant difference was found in the slow component of VO2 between conditions in the first (NaHCO3, 210 +/- 69 ml; placebo, 239 +/- 105 ml) or second trial (NaHCO3, 123 +/- 88 ml; placebo, 197 +/- 101 ml). In conclusion, pre exercise NaHCO3 ingestion did not significantly attenuate the VO2 slow component of professional road cyclists during high-intensity exercise. PMID- 12587891 TI - Lunge performance and its determinants. AB - For activities such as squash, badminton and fencing, the ability to quickly complete a lunge and return to the start or move off in another direction is critical for success. Determining which strength qualities are important predictors of lunge performance was the focus of this study. Thirty-one male athletes performed: (1) a unilateral maximal squat (one-repetition maximum, 1-RM) and unilateral jump squat (50% 1-RM) on an instrumented supine squat machine, and (2) a forward lunge while attached to a linear transducer. We performed stepwise multiple regression analysis with lunge performance as the dependent variable and various strength, flexibility and anthropometric measures as the independent variables. From the many strength and power measures calculated, time to peak force was the best single predictor of lunge performance, which accounted for 55% of the explained variance. The best three-variable model for predicting lunge performance accounted for 76-85% of the explained variance. The models differed, however, according to whether lunge performance was expressed relative to body mass (time to peak force, mean power and relative strength = 76%) or taken as an absolute value (time to peak force, leg length and flexibility = 85%). We conclude that one to two trials were reliable for strength diagnosis and that one strength measure cannot accurately explain functional performance because other factors, such as body mass, flexibility and leg length, have diverse effects on the statistical models. PMID- 12587892 TI - The effects of bungy weight training on muscle function and functional performance. AB - Eccentric strength training is thought to be important for improving functional performance. A form of training that may enhance the eccentric training stimulus is the attachment of a rubber bungy to the strength-training apparatus in such a way that the return velocity and, therefore, the force required to decelerate the load at the end of the eccentric phase are increased. To determine the effects of elastic bungy training, we performed two studies. In the first, we examined the electromyographic (EMG) and kinematic characteristics of three different squat techniques: traditional squat, non-bungy jump squat and bungy jump squat. In the second study, we examined whether jump squat training with and without the attachment of a rubber bungy to an isoinertial supine squat machine affects muscle function, multidirectional agility, lunge ability and single leg jump performance. The EMG activity of the vastus lateralis and gastrocnemius muscles was recorded. An instrumented isoinertial supine squat machine was used to measure maximal strength and various force, velocity and power measures in both studies. Participants were randomly assigned to one of three groups: a control group and two weight-trained groups, one of which performed bungy squat jumps and one of which performed non-bungy squat jumps. The two experimental groups performed 10 weeks of ballistic weight training. The kinematic and EMG characteristics of the bungy and non-bungy squat techniques differed significantly from those of the traditional squat on all the variables measured. The only difference between the bungy squat and non-bungy squat training was greater EMG activity during the later stages (70-100%) of the eccentric phase of the bungy squat condition. The 10 weeks of bungy squat and non-bungy squat jump weight training were found to be equally effective in producing improvements in a variety of concentric strength and power measures (10.6-19.8%). These improvements did not transfer to improved performance for the single leg jump and multidirectional agility. However, bungy weight training did lead to a significant improvement in lunge performance (21.5%) compared with the other groups. PMID- 12587894 TI - Automatic activation of multiplication facts: evidence from the nodes adjacent to the product. AB - Adult observers are widely assumed to be equipped with a specific memory store containing arithmetic facts. The present study was aimed at exploring the possibility of obtaining an automatic activation of multiplication facts by using the number-matching paradigm (LeFevre, Bisanz, & Mrkonjic, 1988), in which mental arithmetic is task irrelevant. In particular, we were interested in exploring whether the nodes that precede or follow the product node in the multiplication table can also be automatically activated as a consequence of the mere presentation of two numbers. In Experiments 1 and 2, we showed that participants were slower in responding "no" to probes that were numbers adjacent to the product in the table related to the first operand of the initial pair than to probes that were unrelated to the initial pair. In Experiments 3 and 4, we showed a similar pattern for probes that were numbers adjacent to the product in the table related to the second operand of the initial pair. Experiments 5 and 6 ruled out alternative accounts and confirmed the results of the previous experiments. Taken together the present findings suggest that multiplication facts are stored in a highly related network in which activation spreads from the product node to adjacent nodes. PMID- 12587893 TI - Comprehension of algebraic expressions by experienced users of mathematics. AB - Little is known about how people comprehend mathematical expressions. In the present study we investigate the internal representations used by experienced users of mathematics to encode algebraic expressions. Initially, a memory recognition task was conducted that examined the role of mathematical syntax in the encoding of algebraic expressions. The results indicate that participants could more readily identify those parts of a previously seen expression that were syntactically well formed than those that were not well formed, suggesting that syntax plays an important role. To determine the level of syntactic structure involved, a second recognition task was conducted. The results indicate that algebraic expressions are encoded into components that represent the phrasal constituents of the expression. However, the results of these experiments did not rule out the possibility that the visual processes of perceptual organization were used to encode the expressions, or that the data were a consequence of a mathematical "lexicon" of common mathematical "phrases". Three further experiments were conducted to examine this, the results of which indicate that the encoding of algebraic expressions is based primarily on processes that occur beyond the level of visual or "lexical" processing. This is consistent with our finding that syntactic structure plays a key role. PMID- 12587895 TI - The impact of reader skill on phonological processing in visual word recognition. AB - There has been much debate about the role of phonology in reading. This debate has been fuelled, in part, by mixed findings for phonological effects in lexical decision tasks. In the present research we investigated the impact of reader skill on three phonological effects (homophone, homograph, and regularity effects) in a lexical decision task and in a phonological lexical decision task. In both tasks, the more skilled readers showed different patterns of phonological effects from those of the less skilled readers; in particular, less skilled readers showed regularity effects in both tasks whereas more skilled readers did not. We concluded that more skilled readers activate phonology in these tasks but do so more efficiently, with less spurious phonological activation. PMID- 12587896 TI - Phonological information in immediate and delayed sentence recall. AB - Potter and Lombardi (1990) state in their conceptual regeneration hypothesis that immediate sentence recall is only based on conceptual and lexical information; phonological information does not contribute. As experimental evidence for this hypothesis, they reported that if a sentence is followed by a word list that included a lure word similar to one of the content words of the sentence (target word), the lure word frequently intrudes into sentence recall. We demonstrated that Potter and Lombardi did not observe any influence of phonological information because list presentation followed sentence presentation, and phonological information was discarded. We observed that phonological information influenced the intrusion rate if recall was not delayed by the subsequent presentation of a word list. With immediate recall, the lure intrusion effect disappeared in auditorily presented sentences. This shows that, if available, phonological information contributes to sentence recall. PMID- 12587897 TI - The use of control groups in artificial grammar learning. AB - Experimenters assume that participants of an experimental group have learned an artificial grammar if they classify test items with significantly higher accuracy than does a control group without training. The validity of such a comparison, however, depends on an additivity assumption: Learning is superimposed on the action of non-specific variables-for example, repetitions of letters, which modulate the performance of the experimental group and the control group to the same extent. In two experiments we were able to show that this additivity assumption does not hold. Grammaticality classifications in control groups without training (Experiments 1 and 2) depended on non-specific features. There were no such biases in the experimental groups. Control groups with training on randomized strings (Experiment 2) showed fewer biases than did control groups without training. Furthermore, we reanalysed published research and demonstrated that earlier experiments using control groups without training had produced similar biases in control group performances, bolstering the finding that using control groups without training is methodologically unsound. PMID- 12587898 TI - Measuring learning using an untrained control group: comment on R. Reber and Perruchet. AB - R. Reber and Perruchet (this issue) argue that use of control groups without training is unsound for establishing that learning has occurred. We show that inferring learning from a difference between a trained group and an untrained control in no way relies on their implausible additivity assumption, and that untrained control groups can be an invaluable aid to the researcher. PMID- 12587899 TI - Belief bias and relational reasoning. AB - When people evaluate categorical syllogisms, they tend to reject unbelievable conclusions and accept believable ones irrespective of their validity. Typically, this effect is particularly marked for invalid conclusions that are possible, but do not necessarily follow, given the premises. However, smaller believability effects can also be detected for other types of conclusion. Three experiments are reported here, in which an attempt was made to determine whether belief bias effects can manifest themselves on the relational inference task. Subjects evaluated the validity of conclusions such as William the Conqueror was king after the Pyramids were built (temporal task) or Manchester is north of Bournemouth (spatial task) with respect to their premises. All of the major findings for equivalent categorical syllogism tasks were replicated. However, the overall size of the main effect of believability appears to be related to task presentation, a phenomenon not previously identified for categorical syllogisms and which current theories of belief bias have difficulty explaining. PMID- 12587900 TI - Functional representation of 3D space in endogenous attention shifts. AB - The aim of this study was to explore whether the attentional system, as far as an endogenous orienting is concerned, allocates resources along the sagittal plane and whether such a process is affected by, and is likely to be based on, different functional representations of 3D space in the brain. Several models make a main action-based distinction between representations of peripersonal space and of those extrapersonal space. Accordingly, if attention has to move from one representation to another, it should be possible to observe a decrease in performance during such a transition. To test this hypothesis three experiments were run in which participants performed a cued detection task. Cue stimuli were informative and were centrally located around the fixation point. Target stimuli were displayed at four different depth planes. In the first experiment, assuming that the border between the peripersonal space and the extrapersonal space was at 1 m from the observer, half the target stimuli were located in the peripersonal space and half in the extrapersonal space. The fixation point was located at 1 m from the observer. In the second experiment, the fixation point was moved at 2 m from the observer in order to rule out the possible effects of ocular motor programming. In the third experiment, in order to rule out effects related to the spatial layout of target stimuli (i.e., centre of mass effect) two target stimuli were located in the peripersonal space and six in the extrapersonal space. In all the experiments, besides a validity effect, we observed greater reaction times when attention shift was across spatial representations than when it was within the same representation. The implications for action-oriented models of attention are discussed. PMID- 12587901 TI - The effects of delays in radiotherapy treatment on tumour control. AB - There is often a considerable delay from initial tumour diagnosis to the start of radiotherapy treatment, which may be due to factors such as waiting lists and referral delays. This paper uses widely published models and clinical parameters to calculate the effect of delays in treatment on local tumour control for four different types of tumour-squamous cell carcinoma (head and neck), breast, cervix and prostate. The Poisson model for tumour control probability (TCP), an exponential function for tumour growth and the linear quadratic model of cell kill are used to calculate the change in TCP for delays between diagnosis and treatment of up to 100 days. Typical values of the clinical parameters have been taken from the literature; these include alpha and beta, sigma(alpha), tumour size at diagnosis, pre-treatment doubling time, delay in onset of accelerated repopulation and doubling time during treatment. It is acknowledged that there are limitations in the reliability of these data for predicting absolute values of tumour control, but models are still useful for predicting how changes in treatment parameters are likely to affect the outcome. It is shown that for fast growing tumours a delay of 1-2 months can have a significant adverse effect on the outcome, whereas for slow-growing tumours such as Ca prostate a delay of a few months does not significantly reduce the probability of tumour control. These calculations show the importance of ensuring that delays from diagnosis through to treatment are minimized, especially for patients with rapidly proliferating tumours. PMID- 12587902 TI - Dosimetric verification of x-ray fields with steep dose gradients using an electronic portal imaging device. AB - Regions with steep dose gradients are often encountered in clinical x-ray beams, especially with the growing use of intensity modulated radiotherapy (IMRT). Such regions are present both at field edges and, for IMRT, in the vicinity of the projection of sensitive anatomical structures in the treatment field. Dose measurements in these regions are often difficult and labour intensive, while dose prediction may be inaccurate. A dedicated algorithm developed in our institution for conversion of pixel values, measured with a charged coupled device camera based fluoroscopic electronic portal imaging device (EPID), into absolute absorbed doses at the EPID plane has an accuracy of 1-2% for flat and smoothly modulated fields. However, in the current algorithm there is no mechanism to correct for the (short-range) differences in lateral electron transport between water and the metal plate with the fluorescent layer in the EPID. Moreover, lateral optical photon transport in the fluorescent layer is not taken into account. This results in large deviations (>10%) in the penumbra region of these fields. We have investigated the differences between dose profiles measured in water and with the EPID for small heavily peaked fields. A convolution kernel has been developed to empirically describe these differences. After applying the derived kernel to raw EPID images, a general agreement within 2% was obtained with the water measurements in the central region of the fields, and within 0.03 cm in the penumbra region. These results indicate that the EPID is well suited for accurate dosimetric verification of steep gradient x-ray fields. PMID- 12587903 TI - One-dimensional phased array with mechanical motion for conformal ultrasound hyperthermia. AB - This paper investigates the feasibility of conformal heating for external ultrasound hyperthermia by using a phased array transducer with mechanical motion. In this system, a one-dimensional phased array is arranged on a shaft and moves along the shaft, while dynamically focusing on the planning target volume (PTV) with numerous focal spots. To prevent overheating in the intervening tissue between the skin and the PTV, the shaft and the phased array are rotated together to enlarge the acoustical window. With the purpose of conformal heating, the power deposition of the PTV is constructed by combinations of the focal spots and an iterative gradient descent method is then used to determine an optimal set of power weightings for the focal spots. Different tumour shapes are evaluated and the simulation results demonstrate that the volume percentage of the PTV with temperatures higher than 43 degrees C is over 95%. The overheating volume outside the PTV is less than 25% of the PTV. This method provides good conformal heating for external ultrasound hyperthermia. The concept of combining electrical focusing and mechanical motion has the advantages of both enlarging the acoustic window and providing dynamic focusing ability, which is essential for successful conformal heating. PMID- 12587904 TI - Absence of multiple local minima effects in intensity modulated optimization with dose-volume constraints. AB - This paper reports on the analysis of intensity modulated radiation treatment optimization problems in the presence of non-convex feasible parameter spaces caused by the specification of dose-volume constraints for the organs-at-risk (OARs). The main aim was to determine whether the presence of those non-convex spaces affects the optimization of clinical cases in any significant way. This was done in two phases: (1) Using a carefully designed two-dimensional mathematical phantom that exhibits two controllable minima and with randomly initialized beamlet weights, we developed a methodology for exploring the nature of the convergence characteristics of quadratic cost function optimizations (deterministic or stochastic). The methodology is based on observing the statistical behaviour of the residual cost at the end of optimizations in which the stopping criterion is progressively more demanding and carrying out those optimizations to very small error changes per iteration. (2) Seven clinical cases were then analysed with dose-volume constraints that are stronger than originally used in the clinic. The clinical cases are two prostate cases differently posed, a meningioma case, two head-and-neck cases, a spleen case and a spine case. Of the 14 different sets of optimizations (with and without the specification of maximum doses allowed for the OARs), 12 fail to show any effect due to the existence of non-convex feasible spaces. The remaining two sets of optimizations show evidence of multiple minima in the solutions, but those minima are very close to each other in cost and the resulting treatment plans are practically identical, as measured by the quality of the dose-volume histograms (DVHs). We discuss the differences between fluence maps resulting from those similar treatment plans. We provide a possible reason for the observed results and conclude that, although the study is necessarily limited, the annealing characteristics of a simulated annealing method may not be justified in clinical optimization in the presence of dose-volume constraints. The results of optimizations by the Newton gradient (NG) method with a quadratic cost function are reported in detail. An adaptive simulated annealing method, optimizing the same function, and the dynamically penalized likelihood method, optimizing a log likelihood function, have also been used in the study. The results of the latter two methods have only been discussed briefly, as they yielded the same conclusions as the NG method. PMID- 12587905 TI - Stereotactic imaging for radiotherapy: accuracy of CT, MRI, PET and SPECT. AB - CT, MRI, PET and SPECT provide complementary information for treatment planning in stereotactic radiotherapy. Stereotactic correlation of these images requires commissioning tests to confirm the localization accuracy of each modality. A phantom was developed to measure the accuracy of stereotactic localization for CT, MRI, PET and SPECT in the head and neck region. To this end. the stereotactically measured coordinates of structures within the phantom were compared with their mechanically defined coordinates. For MRI, PET and SPECT, measurements were performed using two different devices. For MRI, T1- and T2 weighted imaging sequences were applied. For each measurement, the mean radial deviation in space between the stereotactically measured and mechanically defined position of target points was determined. For CT, the mean radial deviation was 0.4 +/- 0.2 mm. For MRI, the mean deviations ranged between 0.7 +/- 0.2 mm and 1.4 +/- 0.5 mm, depending on the MRI device and the imaging sequence. For PET, mean deviations of 1.1 +/- 0.5 mm and 2.4 +/- 0.3 mm were obtained. The mean deviations for SPECT were 1.6 +/- 0.5 mm and 2.0 +/- 0.6 mm. The phantom is well suited to determine the accuracy of stereotactic localization with CT, MRI, PET and SPECT in the head and neck region. The obtained accuracy is well below the physical resolution for CT, PET and SPECT, and of comparable magnitude for MRI. Since the localization accuracy may be device dependent, results obtained at one device cannot be generalized to others. PMID- 12587906 TI - MRI-guided gas bubble enhanced ultrasound heating in in vivo rabbit thigh. AB - In this study, we propose a focused ultrasound surgery protocol that induces and then uses gas bubbles at the focus to enhance the ultrasound absorption and ultimately create larger lesions in vivo. MRI and ultrasound visualization and monitoring methods for this heating method are also investigated. Larger lesions created with a carefully monitored single ultrasound exposure could greatly improve the speed of tumour coagulation with focused ultrasound. All experiments were performed under MRI (clinical, 1.5 T) guidance with one of two eight-sector, spherically curved piezoelectric transducers. The transducer, either a 1.1 or 1.7 MHz array, was driven by a multi-channel RF driving system. The transducer was mounted in an MRI-compatible manual positioning system and the rabbit was situated on top of the system. An ultrasound detector ring was fixed with the therapy transducer to monitor gas bubble activity during treatment. Focused ultrasound surgery exposures were delivered to the thighs of seven New Zealand while rabbits. The experimental, gas-bubble-enhanced heating exposures consisted of a high amplitude 300 acoustic watt, half second pulse followed by a 7 W, 14 W or 21 W continuous wave exposure for 19.5 s. The respective control sonications were 20 s exposures of 14 W, 21 W and 28 W. During the exposures, MR thermometry was obtained from the temperature dependency of the proton resonance frequency shift. MRT2-enhanced imaging was used to evaluate the resulting lesions. Specific metrics were used to evaluate the differences between the gas-bubble-enhanced exposures and their respective control sonications: temperatures with respect to time and space, lesion size and shape, and their agreement with thermal dose predictions. The bubble-enhanced exposures showed a faster temperature rise within the first 4 s and higher overall temperatures than the sonications without bubble formation. The spatial temperature maps and the thermal dose maps derived from the MRI thermometry closely correlated with the resulting lesion as examined by T2-weighted imaging. The lesions created with the gas-bubble-enhanced heating exposures were 2-3 times larger by volume, consistently more spherical in shape and closer to the transducer than the control exposures. The study demonstrates that gas bubbles can reliably be used to create significantly larger lesions in vivo. MRI thermometry techniques were successfully used to monitor the thermal effects mediated by the bubble-enhanced exposures. PMID- 12587907 TI - Methods for measuring the infrared spectra of biological cells. AB - Infrared (IR) spectroscopy of biological cells is a growing area of research, with many papers focusing on differences between the spectra of cancerous and noncancerous cells. Much of this research has been performed using a monolayer of dehydrated cells. We posit that the use of monolayers can introduce artefacts that lead to an apparent but inaccurate measurement of differences between cancerous and noncancerous cells. Additionally, the use of dried cells complicates the extraction of biochemical information from the IR spectra. We demonstrate that using suspensions of viable cells in aqueous suspension reduces measurement artefacts and facilitates determining the concentration of the major biochemical components via a linear least-squares fit of the component spectra to the spectrum of the cells. PMID- 12587908 TI - The energy-dependent electron loss model: backscattering and application to heterogeneous slab media. AB - Electron backscattering has been incorporated into the energy-dependent electron loss (EL) model and the resulting algorithm is applied to predict dose deposition in slab heterogeneous media. This algorithm utilizes a reflection coefficient from the interface that is computed on the basis of Goudsmit-Saunderson theory and an average energy for the backscattered electrons based on Everhart's theory. Predictions of dose deposition in slab heterogeneous media are compared to the Monte Carlo based dose planning method (DPM) and a numerical discrete ordinates method (DOM). The slab media studied comprised water/Pb, water/Al, water/bone, water/bone/water, and water/lung/water, and incident electron beam energies of 10 MeV and 18 MeV. The predicted dose enhancement due to backscattering is accurate to within 3% of dose maximum even for lead as the backscattering medium. Dose discrepancies at large depths beyond the interface were as high as 5% of dose maximum and we speculate that this error may be attributed to the EL model assuming a Gaussian energy distribution for the electrons at depth. The computational cost is low compared to Monte Carlo simulations making the EL model attractive as a fast dose engine for dose optimization algorithms. The predictive power of the algorithm demonstrates that the small angle scattering restriction on the EL model can be overcome while retaining dose calculation accuracy and requiring only one free variable, chi, in the algorithm to be determined in advance of calculation. PMID- 12587909 TI - Light propagation in dentin: influence of microstructure on anisotropy. AB - We investigated the dependence of light propagation in human dentin on its microstructure. The main scatterers in dentin are the tubules, the shape of which can be approximated as long cylinders. We calculated the scattering of electromagnetic waves by an infinitely long cylinder and applied the results in a Monte Carlo code that simulates the light propagation in a dentin slab considering multi-scattering. The theory was compared with goniometric measurements. A pronounced anisotropic scattering pattern was found experimentally and theoretically. In addition, intensity peaks were measured which are shown to be caused by light diffraction by the tubules. PMID- 12587910 TI - Spectra from 2.5-15 microm of tissue phantom materials, optical clearing agents and ex vivo human skin: implications for depth profiling of human skin. AB - Infrared measurements have been used to profile or image biological tissue, including human skin. Usually, analysis of such measurements has assumed that infrared absorption is due to water and collagen. Such an assumption may be reasonable for soft tissue, but introduction of exogenous agents into skin or the measurement of tissue phantoms has raised the question of their infrared absorption spectrum. We used Fourier transform infrared spectroscopy in attenuated total reflection mode to measure the infrared absorption spectra, in the range of 2-15 microm, of water, polyacrylamide, Intralipid, collagen gels, four hyperosmotic clearing agents (glycerol, 1,3-butylene glycol, trimethylolpropane, Topicare), and ex vivo human stratum corneum and dermis. The absorption spectra of the phantom materials were similar to that of water, although additional structure was noted in the range of 6-10 microm. The absorption spectra of the clearing agents were more complex, with molecular absorption bands dominating between 6 and 12 microm. Dermis was similar to water, with collagen structure evident in the 6-10 microm range. Stratum corneum had a significantly lower absorption than dermis due to a lower content of water. These results suggest that the assumption of water-dominated absorption in the 2.5-6 microm range is valid. At longer wavelengths, clearing agent absorption spectra differ significantly from the water spectrum. This spectral information can be used in pulsed photothermal radiometry or utilized in the interpretation of reconstructions in which a constant mu(ir) is used. In such cases, overestimating mu(ir) will underestimate chromophore depth and vice versa, although the effect is dependent on actual chromophore depth. PMID- 12587911 TI - Estimation of a radiation time prolongation factor for intensity-modulated radiotherapy. AB - In general, the deposition of a given target dose requires a longer radiation time for intensity-modulated photon beams (IMBs) than for unmodulated beams. Hence, the routine use of intensity-modulated radiotherapy (IMRT) has repercussions both on the exposure of the patient to scatter and institutional radiation safety. A rule of thumb is presented to assess the maximum prolongation of radiation time for a case class in an idealized setting using static superimposed field segments. The method considers only the degree to which risk structures have to be blocked to meet specified dose restrictions. PMID- 12587912 TI - On biologically conformal boost dose optimization. AB - A method is described that allows the inclusion of biological imaging data in the optimization of intensity-modulated radiotherapy to produce dose boosts that conform with target subvolumes of potentially reduced radiosensitivity. The biological image (e.g. PET, fMRI, etc) is transformed into a dose efficiency distribution using a piecewise linear calibration function with a prescribed maximum boost factor. Instead of dose alone, the cost function of the optimization algorithm depends on the product of the physical dose times dose efficiency. An example case of a base-of-tongue tumour which was imaged with the hypoxia tracer fluoro-misonidazole is presented, showing the excellent capability of IMRT to produce dose distributions that conform to spatially variable dose prescriptions. PMID- 12587913 TI - The traps and pitfalls inherent in the correlation of changes in the fibre diffraction pattern of hair with breast cancer. AB - After a correlation between breast cancer and changes in the fibre x-ray diffraction of hair was first published, a number of groups reported their failure to reproduce this phenomenon. There is evidence that these failures resulted from either diffraction images of insufficient quality or analysis techniques unable to distinguish features resulting from the presence of breast cancer from those resulting from mechanical damage. Using an independent analysis of the same dataset reported by one of these unsuccessful groups, the author demonstrates some of the critical flaws inherent in this type of intensity-based analysis. PMID- 12587914 TI - Patterning the limb before and after SHH signalling. AB - The vertebrate limb is one of the most relevant experimental models for analysing cell-cell signalling during patterning of embryonic fields and organogenesis. Recently, the combination of molecular and genetic studies with experimental manipulation of developing limb buds has significantly advanced our understanding of the complex molecular interactions co-ordinating limb bud outgrowth and patterning. Some of these studies have shown that there is a need to revise some of the textbook views of vertebrate limb development. In this review, we discuss how signalling by the polarizing region is established and how limb bud morphogenesis is controlled by both long-range and signal relay mechanisms. We also discuss recent results showing that differential mesenchymal responsiveness to SHH signalling is established prior to its expression by the polarizing region. PMID- 12587915 TI - Sonic hedgehog: restricted expression and limb dysmorphologies. AB - Sonic hedgehog, SHH, is required for patterning the limb. The array of skeletal elements that compose the hands and feet, and the ordered arrangement of these bones to form the pattern of fingers and toes are dependent on SHH. The mechanism of action of SHH in the limb is not fully understood; however, an aspect that appears to be important is the localized, asymmetric expression of Shh. Shh is expressed in the posterior margin of the limb bud in a region defined as the zone of polarizing activity (ZPA). Analysis of mouse mutants which have polydactyly (extra toes) shows that asymmetric expression of Shh is lost due to the appearance of an ectopic domain of expression in the anterior limb margin. One such polydactylous mouse mutant, sasquatch (Ssq), maps to the corresponding chromosomal region of the human condition pre-axial polydactyly (PPD) and thus represents a model for this condition. The mutation responsible for Ssq is located 1 Mb away from the Shh gene; however, the mutation disrupts a long-range cis-acting regulator of Shh expression. By inference, human pre-axial polydactyly results from a similar disruption of Shh expression. Other human congenital abnormalities also map near the pre-axial polydactyly locus, suggesting a major chromosomal region for limb dysmorphologies. The distinct phenotypes range from loss of all bones of the hands and feet to syndactyly of the soft tissue and fusion of the digits. We discuss the role played by Shh expression in mouse mutant phenotypes and the human limb dysmorphologies. PMID- 12587916 TI - Developmental mechanisms underlying polydactyly in the mouse mutant Doublefoot. AB - The pre-axial polydactylous mouse mutant Doublefoot has 6-9 digits per limb but lacks anteroposterior polarity (there is no biphalangeal digit 1). It differs from other polydactylous mutants in showing normal Shh expression, but polarizing activity (shown by mouse-chick grafting experiments) and hedgehog signalling activity (shown by expression of Ptc1) are present throughout the distal mesenchyme. The Dbf mutation has not yet been identified. Here we review current understanding of this mutant, and briefly report new results indicating (1) that limb bud expansion is concomitant with ectopic lhh expression and with extension of the posterior high cell proliferation rate into the anterior region, and (2) that the Dbf mutation is epistatic to Shh in the limb. PMID- 12587917 TI - Why study human limb malformations? AB - Congenital limb malformations occur in 1 in 500 to 1 in 1000 human live births and include both gross reduction defects and more subtle alterations in the number, length and anatomy of the digits. The major causes of limb malformations are abnormal genetic programming and intra-uterine disruption to development. The identification of causative gene mutations is important for genetic counselling and also provides insights into the mechanisms controlling limb development. This article illustrates some of the lessons learnt from the study of human limb malformation, organized into seven categories. These are: (1) identification of novel genes, (2) allelic mutation series, (3) pleiotropy, (4) qualitative or (5) quantitative differences between mouse and human development, (6) physical and teratogenic disruption, and (7) unusual biological phenomena. PMID- 12587918 TI - Congenital talipes equinovarus (clubfoot): a disorder of the foot but not the hand. AB - Idiopathic (non-syndromic) congenital talipes equinovarus, or clubfoot, is a poorly understood but common developmental disorder of the lower limb, which affects at least 2 per 1000 Scottish births (ISD data). It is defined as a fixation of the foot in a hand-like orientation--in adduction, supination and varus--with concomitant soft tissue abnormalities. Despite advances in treatment, disability often persists. The aetiology of the condition has been little studied and is poorly understood. Neurological, muscular, bony, connective tissue and vascular mechanisms have been proposed, but the only firm evidence is that the mildest cases appear to be associated with intra-uterine posture. There is evidence for a genetic contribution to congenital talipes equinovarus aetiology. Its incidence varies with ethnic group, and we found that a family history is present in 24-50% of cases, depending on the population studied. Complex segregation analysis suggests that the most likely inheritance pattern is a single gene of major effect operating against a polygenic background. Possible mechanisms for congenital talipes equinovarus are discussed. PMID- 12587919 TI - Epithelial mesenchymal interactions, the ECM and limb development. AB - It has been long since recognized that cellular interactions are not always direct, i.e. they do not always take place between cells contacting each other, or between cells that emit soluble factors and other cells, which respond to it. In contrast, cross-talk between cells is frequently based on signals attached to the extracellular matrix (ECM). Thus besides proximate cell-to-cell contact, certain interactions are mediated by the ECM in a sequence: cell-to-matrix, matrix-to-cell. ECM-mediated interactions may take place within a group or sheet of cells or across adjacent cell sheets. A modified mat-like ECM, the basement membrane, separates adjacent cell sheets and mediates their interactions. Since cell sheets separated by basement membranes are an elementary feature of metazoan histology, interactions with the basement membrane have considerable importance. Recently accumulated evidence emphasizes the importance of ECM-mediated interactions. It is becoming increasingly evident that the ECM functions not only as an architectural component, but it is involved also in signal transduction. This evidence derives from four main sources: from the structure of receptor ligand complexes, from Drosophila and C elegans genetics, from cell biological observations and from the analysis of mammalian development. In this review, I will touch upon recent evidence, illustrated by examples of FGF signalling in vertebrate limb development. Although the involvement of matrix components is not yet proven for all cases directly, the strength of multiple indications suggests that a better understanding of ECM-mediated interactions will shed new light on cell differentiation. PMID- 12587920 TI - Digital development and morphogenesis. AB - Signalling interactions between the polarizing region, which produces SHH, and the apical ectodermal ridge, which produces FGFs, are essential for outgrowth and patterning of vertebrate limbs. However, mechanisms that mediate translation of early positional information of cells into anatomy remain largely unknown. In particular, the molecular and cellular basis of digit morphogenesis are not fully understood, either in terms of the formation of the different digits along the antero-posterior axis or in the way digits stop growing once pattern formation has been completed. Here we will review recent data about digit development. Manipulation of morphogenetic signals during digit formation, including application of SHH interdigitally, has shown that digit primordia possess a certain plasticity, and that digit anatomy becomes irreversibly fixed during morphogenesis. The process of generation of joints and thus segmentation and formation of digit tips is also discussed. PMID- 12587921 TI - The formation of skeletal muscle: from somite to limb. AB - During embryogenesis, skeletal muscle forms in the vertebrate limb from progenitor cells originating in the somites. These cells delaminate from the hypaxial edge of the dorsal part of the somite, the dermomyotome, and migrate into the limb bud, where they proliferate, express myogenic determination factors and subsequently differentiate into skeletal muscle. A number of regulatory factors involved in these different steps have been identified. These include Pax3 with its target c-met, Lbx1 and Mox2 as well as the myogenic determination factors Myf5 and MyoD and factors required for differentiation such as Myogenin, Mrf4 and Mef2 isoforms. Mutants for genes such as Lbx1 and Mox2, expressed uniformly in limb muscle progenitors, reveal unexpected differences between fore and hind limb muscles, also indicated by the differential expression of Tbx genes. As development proceeds, a secondary wave of myogenesis takes place, and, postnatally, satellite cells become located under the basal lamina of adult muscle fibres. Satellite cells are thought to be the progenitor cells for adult muscle regeneration, during which similar genes to those which regulate myogenesis in the embryo also play a role. In particular, Pax3 as well as its orthologue Pax7 are important. The origin of secondary/fetal myoblasts and of adult satellite cells is unclear, as is the relation of the latter to so-called SP or stem cell populations, or indeed to potential mesangioblast progenitors, present in blood vessels. The oligoclonal origin of postnatal muscles points to a small number of founder cells, whether or not these have additional origins to the progenitor cells of the somite which form the first skeletal muscles, as discussed here for the embryonic limb. PMID- 12587923 TI - The innervation of FGF-induced additional limbs in the chick embryo. AB - Motoneurones that supply the vertebrate limb innervate their muscle targets in a highly reproducible manner. As development proceeds, these limb-specific motoneurones send out axons, which grow towards the developing limb and then congregate at its base to form the plexus. In the plexus, in response to unknown positional cues, these axons rearrange, often changing their original spatial relationships, before sorting out to emerge in the defined nerve trunks that innervate the limb. Several proposals have been put forward to explain how this reproducible innervation pattern is achieved. These include (1) that early differences in the motoneurone identity dictate their future axonal trajectories, (2) that axons actively respond to attractive or repulsive positional cues provided by the limb bud itself, or (3) that motor axons are passively deployed, following pathways of least mechanical resistance. We have addressed the question of the relative roles of motoneurone identity and the signals that the axons encounter on their journey towards the limb bud. Using the developing chick embryo as our experimental model we tested the effect of providing an additional limb target for motor axons leaving the flank level of the spinal cord. To do this we placed FGF-soaked beads in the presumptive flank of 2-day-old chick embryos. This treatment induces an additional limb containing muscles. We investigated whether such additional limbs are innervated and by which neurones. We show that rather than the additional limbs being solely supplied by axons diverted from the two existing limb plexuses, motoneurones that normally supply the flank alter their trajectories to enter the induced limb. Once in the limb, axons respond to positional cues within the bud to generate the stereotypical innervation pattern. Our results show that the tendency of 'flank' motoneurones to innervate flank can be overcome by the presence of an additional limb. PMID- 12587922 TI - Regulation of myogenic differentiation in the developing limb bud. AB - The limb myogenic precursors arise by delamination from the lateral dermomyotome in response to signals from the lateral plate mesoderm. They subsequently migrate into the developing limb bud where they switch on the expression of the myogenic regulatory factors, MyoD and Myf5, and coalese to form the dorsal and ventral muscle masses. The myogenic cells subsequently undergo terminal differentiation into slow or fast fibres which have distinct contractile properties determining how a muscle will function. In general, fast fibres contract rapidly with high force and are characterized by the expression of fast myosin heavy chains (MyHC). These fibres are needed for movement. In contrast, slow fibres express slow MyHC, contract slowly and are required for maintenance of posture. This review focuses on the molecular signals that control limb myogenic development from the initial delamination and migration of the premyogenic cells to the ultimate formation of the complex muscle pattern and differentiation of slow and fast fibres. PMID- 12587925 TI - Molecular control of arterial-venous blood vessel identity. AB - Recent research has demonstrated that not only haemodynamic factors but also genetic programmes control arterial-venous cell fate and blood vessel identity. The identification of arteries and veins was previously based solely on morphological criteria and is now greatly facilitated by specific molecular markers. Moreover, signalling pathways controlling the arterial-venous decision during embryonic development have been outlined for the first time. This review gives an up-to-date overview of differentially expressed genes and the regulatory processes leading to the differentiation of arteries and veins. PMID- 12587924 TI - Vascularization of the developing chick limb bud: role of the TGFbeta signalling pathway. AB - The developing vertebrate limb has fascinated developmental biologists and theoreticians for decades as a model system for investigating cell fate, cell signalling and tissue interactions. We are beginning to understand the mechanisms and signalling pathways that control and regulate the outgrowth and formation of the limb bud into a differentiated identifiable limb by late embryogenesis. However, the mechanisms underlying the development and maintenance of the vasculature of the developing limb are far from being completely understood. The vasculature supplies oxygen, nutrients and signals to developing tissues, allowing them to develop and grow. Moreover, a lot of evidence recently points to molecules involved in morphological development also controlling vascular development. Thus understanding how the vasculature forms and is patterned in the developing limb may further our understanding of limb development. In this review I outline how blood vessels are formed and maintained and how the developing chick limb is vascularized. I also review the role of the TGFbeta superfamily signalling pathway in the development of the chick limb vasculature: in particular, how antagonizing TGFbeta signalling in the developing chick limb has shed new light on the role vascular smooth muscle cells play in vessel calibre control and how this work has added to our understanding of TGFbeta superfamily signal transduction. PMID- 12587926 TI - Covering the limb--formation of the integument. AB - An organism's outermost covering, the integument, has evolved to fulfil a diverse range of functions. Skin provides a physical barrier, an environment for immunological surveillance, and also performs a range of sensory, thermoregulatory and biosynthetic functions. Examination of the skin of limb digits reveals a range of skin types including the thickened hairless epidermis of the toe pads (palmar or plantar epidermis) and thinner epidermis between the hair follicles (interfollicular epidermis) of hairy skin. An important developmental function of skin is to give rise to a diverse group of appendages including hair follicles, with associated sebaceous glands (or feathers and scales in chick), eccrine sweat glands and the nail. A key question is how does this morphological variety arise from the single-layered epithelium covering embryonic limb buds? This review will attempt to address this question by linking the extensive morphological/anatomical data on maturation of epidermis and its appendages with (1) current research into the range, plasticity and location of the putative epidermal stems cells; (2) molecular/microenvironmental regulation of epidermal stem cell lineages and lineage choice; and (3) regulation of the differentiation pathways, focusing on differentiation of the interfollicular epidermis. PMID- 12587927 TI - Mouse models for human hair loss disorders. AB - The outer surface of the hand, limb and body is covered by the epidermis, which is elaborated into a number of specialized appendages, evolved not only to protect and reinforce the skin but also for social signalling. The most prominent of these appendages is the hair follicle. Hair follicles are remarkable because of their prolific growth characteristics and their complexity of differentiation. After initial embryonic morphogenesis, the hair follicle undergoes repeated cycles of regression and regeneration throughout the lifetime of the organism. Studies of mouse mutants with hair loss phenotypes have suggested that the mechanisms controlling the hair cycle probably involve many of the major signalling molecules used elsewhere in development, although the complete pathway of hair follicle growth control is not yet understood. Mouse studies have also led to the discovery of genes underlying several human disorders. Future studies of mouse hair-loss mutants are likely to benefit the understanding of human hair loss as well as increasing our knowledge of mechanisms controlling morphogenesis and tumorigenesis. PMID- 12587928 TI - Genetic disorders of palm skin and nail. AB - The outer part of the skin, the epidermis, is specialized to protect the human body from its environment. Because of the high levels of physical stress experienced by the human hand in everyday use, the epidermis of the hand is especially toughened. In particular, the epidermis of the palm is highly specialized to resist mechanical trauma. Like the epidermis, the nails are composed of specialized epithelial cells and are especially strong. In recent years it has become apparent that the physical strength of epithelial cells comes from the keratin cytoskeleton--a dense meshwork of filaments extending throughout the cytoplasm. Keratins are a large family of intermediate filament proteins encoded by more than 50 distinct genes in humans. These different keratin genes are expressed in well-defined combinations in specific epithelial tissues. Several keratin genes are expressed in palmoplantar epidermis and in the stratified epithelia of the nail bed. Genetic mutations in these genes lead to fragility of these tissues and result in a range of genetic disorders characterized by blistering and thickening of palm and sole skin and/or nails. Study of these diseases has shed new light on the vital structural role of keratins in maintaining the integrity of epithelial cells. PMID- 12587929 TI - Postnatal development of the fore- and hindlimbs in the grey short-tailed opossum, Monodelphis domestica. AB - Marsupials are good experimental animals for developmental studies as their offspring are born at a stage comparable to embryonic stages of eutherian species. The South American opossum, Monodelphis domestica, is particularly useful because of its small size and easy maintenance. This study was carried out to compare development of opossum fore- and hindlimbs during postnatal life, using light microscopy and whole mount alizarin staining. At birth, well developed mobile forelimbs show cartilage models of bones and myotubular striated muscle fibres. However, hindlimbs are relatively underdeveloped paddle-like outgrowths. Two days later mesodermal condensations form models of the future hindlimb bones and mononucleate myoblast aggregates are present; by 6 days post partum (dpp) the hindlimb has reached a stage of development similar to that of the forelimb at birth. At this stage, periosteal buds have invaded forelimb long bones and nuclei in forelimb muscle fibres have become displaced to the periphery. The 16 dpp hindlimb shows long bones invaded by periosteal buds and closely packed, striated muscle fibres. Epiphyseal plates are now seen in the forelimb long bones and forelimb muscle fibres show mature characteristics. Musculoskeletal development is well correlated with the functional demands of the limbs during postnatal development in the opossum, which provides an excellent model for investigations into the genes and molecules controlling limb development. PMID- 12587930 TI - Tendon morphogenesis in the developing avian limb: plasticity of fetal tendon fibroblasts. AB - The differentiation and patterning of tendon fibroblasts is at present a poorly understood aspect of musculoskeletal development in the vertebrate limb. Precursors of tendon fibroblasts originate in the somatic mesoderm adjacent to the early limb bud and gradually become incorporated into the limb mesenchyme as development proceeds. It is unclear whether these progenitor cells are committed to the tendon lineage at this early stage, or whether cells become committed only as they are incorporated into a developing tendon. Following a review of our current knowledge of early tendon development, we present recent results from our preliminary studies looking at tendon cell commitment. Using a lacZ encoding replication-deficient retrovirus, we have mapped regions of the early limb bud that contain presumptive tendon progenitor cells, and later used these sites for implanting labelled fetal tendon fibroblasts into developing limbs. Following implantation, we found that these cells successfully re-incorporated into developing proximal and distal tendons, but also surprisingly contributed to other tissue lineages within the limb. Our results suggest that fetal tendon fibroblasts may not be irreversibly committed to a tendon cell fate in the limb and may be somewhat plastic in their ability to integrate into other tissue lineages during development. PMID- 12587931 TI - Evolution of the human hand: the role of throwing and clubbing. AB - It has been proposed that the hominid lineage began when a group of chimpanzee like apes began to throw rocks and swing clubs at adversaries, and that this behaviour yielded reproductive advantages for millions of years, driving natural selection for improved throwing and clubbing prowess. This assertion leads to the prediction that the human hand should be adapted for throwing and clubbing, a topic that is explored in the following report. It is shown that the two fundamental human handgrips, first identified by J. R. Napier, and named by him the 'precision grip' and 'power grip', represent a throwing grip and a clubbing grip, thereby providing an evolutionary explanation for the two unique grips, and the extensive anatomical remodelling of the hand that made them possible. These results are supported by palaeoanthropological evidence. PMID- 12587932 TI - Female sexual dysfunction: the BMJ fails to educate and fails to debate. PMID- 12587933 TI - Abciximab: IIb or not IIb, is that still the question? PMID- 12587934 TI - Clinical guidelines versus clinical practice in the management of low back pain. AB - To date, there have been limited data on the implementation of evidenced-based clinical guidelines for low back pain (LBP). The aim of this study was to assess current management of LBP and evaluate to what extent clinical practice now reflects clinical guidelines. This survey involved the collection and analysis of data from the records of 200 patients who had been referred to a large teaching hospital with LBP Analysis indicated a high use of X-rays, with little evidence of initial biopsychosocial assessment. The most popular treatments were advice, active exercises and McKenzie therapy. Manipulation was rarely used. Overall, a low use of electrotherapy was recorded. The results emphasise how little the clinical guidelines have influenced the decisions of clinicians, and highlight the need to address the barriers to adopting an evidence-based approach in this area. PMID- 12587935 TI - Orbital cellulitis: a review of 21 cases from Ibadan, Nigeria. AB - This study was set up to review cases of orbital cellulitis admitted to the eye ward of our hospital between 1994 and 1998, and to compare the findings with similar studies. Twenty-one (14 males, 7 females, age range 45 days to 50 years [mean 13 years]) of 35 patients with an initial diagnosis of orbital cellulitis were reviewed (14 patients were excluded from the final analysis). Most of the cases were associated with pansinusitis, the commonest organism isolated being Staphylococcus aureus. All the patients were co-managed with ortorhinolaryngologists, and sometimes also with bacteriologists and paediatricians. Seventy-five per cent of the patients made a full recovery. Complications included orbital abscess (28.6%), optic atrophy (10%), cornea opacities (10%) and seizures (5%). Orbital cellulitis remains a childhood infection with potentially devastating consequences. There appears to have been a reduction not only in the number of cases admitted to the eye ward but also in the number and severity of complications. Prompt treatment and a multidisciplinary approach to management may be partly responsible for the improved outcome. This approach therefore needs to be encouraged. PMID- 12587936 TI - Thrombocytopenia in patients with HCV-positive chronic hepatitis: efficacy of leucocyte interferon-alpha treatment. AB - Treating patients with HCV-associated thrombocytopenia is a problem, because the pathogenesis of thrombocytopenia is still unclear. We evaluated the clinical and haematological response to leucocyte interferon-alpha in 20 naive patients with chronic hepatitis C and thrombocytopenia (platelet count <140 x 10(9)/l for at least six months) without portal hypertension and/or hypersplenism. They were treated with leucocyte interferon-alpha (3 MU three times per week) for 12 months and followed up for 12 months. Biochemical (ALT) and virological (HCV-RNA) responses were determined. Two patients discontinued treatment because of hyperthyroidism. Of the 18 patients who completed treatment, 12 (66%) showed a biochemical response, 10 of whom (55.5%) also showed a virological response. At the end of follow-up, four patients (22%) showed a complete (biochemical and virological) response. During treatment, platelet counts decreased to less than 10-20% of pretreatment values in most patients. Three of the four patients with a complete response showed a platelet increase during treatment and throughout the follow-up period. In HCV-associated thrombocytopenia leucocyte interferon-alpha is well tolerated and in cases of sustained virological inhibition is able to ameliorate the disease by increasing the platelet count. PMID- 12587937 TI - Epidemiological, clinical and laboratory features of brucellosis: a prospective evaluation of 120 adult patients. AB - This prospective study was carried out in the department of infectious diseases of Gaziantep University between January 1997 and December 1999 to evaluate the epidemiological, clinical and laboratory features of brucellosis in south-eastern Turkey. One hundred and twenty consecutive patients with active brucellosis were enrolled. The commonest way of transmission was ingestion of milk products from diseased animals. Brucella melitensis was isolated in the specimens of 31 (45.5%) of 68 patients. The commonest abnormalities on physical examination were fever (66.6%), hepatomegaly (63.3%) and splenomegaly (56.6%). Osteoarticular involvement was found in 34 patients (28.3%). Fifteen (12.5%) patients had ocular involvement. Hepatitis, orchiepididymitis, pulmonary involvement and meningitis were found in one (0.8%), four (6.8%), three (2.5%) and one (0.8) patient, respectively. The commonest haematological abnormalities were relative lymphomonocytosis (71.6%) and anaemia (36.6%). In conclusion, brucellosis continues to be a common health problem in communities where the consumption of unpasteurised dairy products is common. Since prevention is as important as early diagnosis in reducing the morbidity of brucellosis, we suggest that improving current health policies with additional educational programmes is essential. PMID- 12587938 TI - Normal values for inner intercanthal, interpupillary, and outer intercanthal distances in the Indian population. AB - The purpose of this study was to establish a set of normal values for inner intercanthal distance (IICD), interpupillary distance (IPD) and outer intercanthal distance (OICD) in a normal Indian population aged 3-80 years and to find whether the difference between the values of these parameters is statistically significant. We also aimed to compute a regression equation for calculating these. Two thousand five hundred clinically normal Indians were enrolled in the study. A millimeter scale was used to measure IPD and IICD. OICD was measured from the bar reading of Hertel's exophthalmometer. Statistical methods were used to calculate the mean and standard deviation values for these parameters. T-test was applied to determine whether difference between the mean values for these parameters in both sexes was statistically significant or not. A regression equation was also computed to find the values for these parameters in both sexes. The IICD, IPD and OICD ranges for males were 20-36 mm, 46-70 mm and 76-105 mm, and for females 20-36 mm, 46-75 mm and 71-105 mm, respectively The difference between the mean values for these parameters was statistically significant in some age groups but not in others. Gender did not usually influence the normal values for IICD, IPD and OICD in the Indian population. The normal values of these parameters were usually lower than those observed for other races. A knowledge of normal values in this population subgroup can help in studying orbito-cranial growth patterns, syndrome diagnosis, surgical management of cranio-facial deformities and trauma, and manufacture of spectacles. PMID- 12587939 TI - Quality of life after use of the patient activated atrial defibrillator. AB - The aim of the study was to evaluate the effects of long-term use of the patient activated atrial defibrillator for recurrent persistent atrial fibrillation (AF) on quality of life (QOL). Fifteen patients were implanted with the Medtronic Jewel AF 7250 device (dual chamber atrial and ventricular defibrillator) for AF only. AF recurrences were treated by out-of-hospital patient-activated atrial defibrillation shocks following the self-administration of oral sedation. QOL was assessed at pre-implant and up to one year with SF36, symptom checklist and HADS questionnaires. A total of 238 (median 10) out-of-hospital patient-activated atrial defibrillation shocks were performed. The SF36 demonstrated a trend toward improvement over the 12-month period compared with baseline values. There was no significant change in the symptom frequency or severity scores. Pre-implant levels of both anxiety and depression were within the predefined range of normality (6 +/- 3 and 3 +/- 2, respectively) and no significant change was seen at 6 months (5 +/- 4 and 3 +/- 3) or 12 months post implant (5 +/- 4 and 2 +/- 2, respectively). After one year of follow-up, 13 (87%) patients said they would have the device implanted again (two were undecided). The study demonstrates that patient-activated atrial defibrillation is a well tolerated therapeutic strategy for maintaining sinus rhythm. PMID- 12587940 TI - Inflammation and Alzheimer's disease. AB - Alzheimer's disease (AD) is classified mainly as a neurodegenerative disease. Recent research, however, has shown that inflammatory mechanisms are also associated with AD and that they could have a role in contributing to the pathogenesis of this disease. The evidence is based on histopathological brain studies, laboratory studies of peripheral inflammation and the fact that certain anti-inflammatory drugs could modify the course of AD. There is wide scope for further research using anti-inflammatory therapies in the prevention/treatment of AD, thereby reducing the burden of this widely prevalent condition in the community. PMID- 12587941 TI - How to use PSA to screen for prostate cancer. AB - Prostate cancer screening of asymptomatic men is not recommended by the National Screening Council at present and is not encouraged in the NHS. A number of randomised controlled trials are under way to establish the place of routine screening of asymptomatic men. We report the possible practice of prostate cancer screening with reference to the appropriate age range for screening, how to screen for prostate cancer and how often, and what constitutes an abnormal result that would merit referral to a urologist for a prostate biopsy. PMID- 12587942 TI - Abciximab and atherosclerotic heart disease: use in percutaneous coronary intervention, acute coronary syndromes and acute myocardial infarction. AB - Abciximab irreversibly binds to the glycoprotein IIb/IIIa receptor on both activated and unactivated platelets inhibiting platelet aggregation. It has been studied in a variety of clinical settings including percutaneous coronary intervention (PCI), ST-elevation myocardial infarction, and non ST-elevation acute coronary syndromes. Abciximab has been demonstrated to reduce acute ischaemic events in the setting of percutaneous intervention with both percutaneous transluminal coronary angioplasy and stenting. It has been shown to be particularly effective when used in patients with acute myocardial infarction undergoing primary PCI. The data for its effective use in the medical phase of therapy for patients with acute coronary syndromes, however, is not as consistent. In this article we review the major trials evaluating the use of abciximab in these clinical scenarios compared with placebo and alternative glycoprotein IIb/IIIa inhibitors. PMID- 12587943 TI - Aripiprazole: a review of its pharmacology and clinical use. AB - Atypical antipsychotics generally have a lower propensity for extrapyramidal side effects (EPSE), hyperprolactinaemia and tardive dyskinesia than that associated with typical antipsychotics but may still produce troublesome side-effects, such as weight gain, cardiac rhythm changes and impaired glucose tolerance. Aripiprazole is a new atypical antipsychotic with a unique receptor binding profile that combines partial agonist activity at D2 and 5HT1A receptors with potent antagonism at 5HT2A receptors. Clinical studies in acute schizophrenic relapse, chronic schizophrenia and acute mania show it is robustly more effective than placebo. Once-daily aripiprazole 15-30 mg is as effective as haloperidol 10 mg/day and risperidone 6 mg/day in short-term treatment of schizophrenia and more effective than haloperidol 7-10 mg/day in maintenance of response in chronic schizophrenia. Aripiprazole appears to be well tolerated, with most studies suggesting a frequency of adverse effects similar to placebo. Aripiprazole seems not to cause significant EPSE, hyperprolactinaemia, excessive weight gain or cardiac rhythm disturbance. Limited data suggest that aripiprazole is not associated with impaired glucose tolerance. PMID- 12587944 TI - Upper airway obstruction in a young adult. AB - Acute epiglottitis is a potentially rapidly fatal illness. Although commonly thought of as a disease affecting children, it is increasingly being diagnosed in adults. We report a case of upper airway obstruction due to epiglottitis in a 37 year-old female. The need for early diagnosis because of the risk of a fatal outcome is highlighted. PMID- 12587945 TI - The express study: preliminary results. AB - The phase III trial EXPRESS was a randomised, double-blind comparison of two anticoagulant therapies used as prophylaxis against venous thromboembolism (VTE) during major elective orthopaedic surgery. The investigational treatment was ximelagatran/melagatran (Exanta), a new direct thrombin inhibitor in both its oral and subcutaneously injectable forms; the comparator was the low molecular weight heparin enoxaparin. Results showed ximelagatran/melagatran was highly statistically significantly superior to enoxaparin in preventing VTE with a relative risk reduction in proximal deep vein thrombosis plus pulmonary embolism of 63.3% (p < 0.000002). The relative risk reduction for total VTE was 23.6% (p < 0.0003). There were no differences in clinically important bleeding events (fatal bleeding, critical organ bleeding and bleeding requiring reoperation) but excess bleeding assessed subjectively was more common in the ximelagatran group than in the enoxaparin group. PMID- 12587946 TI - Management of life-threatening epistaxis in a Jehovah's Witness. AB - The religious beliefs of Jehovah's Witnesses forbid them to receive blood or blood products. We report a case of life-threatening epistaxis in a Jehovah's Witness, causing severe anaemia (Hb 2.7 g/dl). The successful management of this case and literature relating to this are discussed. PMID- 12587947 TI - Amphetamine-related acute myocardial infarction due to coronary artery spasm. AB - A 27-year-old man developed acute myocardial infarction after intravenous amphetamine use. A coronary angiogram showed plaques in the mid-portion of the left anterior descending artery which developed coronary artery spasm after administration of intracoronary ergonovine. The findings in this case suggest that these coronary artery plaques played a role in the endothelial dysfunction resulting from amphetamine use, and that induction of coronary arterial spasm was the likely mechanism of amphetamine-related acute myocardial infarction. PMID- 12587948 TI - Neoadjuvant radiotherapy in anorectal malignant melanoma. AB - Primary anorectal malignant melanoma is a rare neoplasm that carries a poor prognosis with a high metastatic potential. A case of a 60-year-old male who was admitted to the emergency service with signs of colonic obstruction forced us to reconsider diagnostic and therapeutic implications in anorectal melanoma. Following urgent decompression with laparoscopic sigmoid loop colostomy, the patient was treated with preoperative radiotherapy and local excision. He has since been free of disease for 30 months. The previously published reports are reviewed and the current therapeutic options are discussed. PMID- 12587949 TI - Late colonic obstruction after barium meal examination in a myxoedematous patient. AB - A case in which inspissated barium was retained and produced sigmoid obstruction after one year is described. The patient was elderly and myxoedematous. Prolonged retention of barium producing a bowel obstruction requiring surgery is a rare complication and, to our knowledge, is only the second case described in which the symptoms of obstruction appeared so late after the examination. PMID- 12587950 TI - Comparative study of donepezil and rivastigmine. PMID- 12587951 TI - Stress-induced lidocaine modification in serum and tissues. AB - The aim of this study is to examine the influence of acute (trauma) and chronic (cold swimming and adjuvant rheumatoid arthritis) stress on lidocaine concentrations in plasma. Forty male Wistar rats were used. The animals were divided into four groups. Group A served as control. Group B underwent mandible osteotomy. Group C was submitted to swimming stress in cold water 4 degrees C for ten minutes daily for 15 minutes, while group D underwent experimental arthritis with Freud's adjuvant. All groups received lidocaine i.m (2.5 mg/kg). Blood samples were collected and FFA (free fatty acid), unbound-lidocaine, albumin and a1-acid glycoprotein concentrations were estimated. Furthermore, the adrenals, heart and liver were isolated. The adrenals' relative weight (adrenal weight/body weight) was assessed, while lidocaine concentrations in the heart and the liver incubation medium were measured by intertechnic a-counter. Lidocaine and FFA levels in serum as well as the adrenal weights demonstrated a significant elevation in stress-groups as compared to the control group. Furthermore, in the stress-groups, lidocaine concentrations in heart tissue were significantly increased, whereas in the liver they were significantly reduced as compared to the control group. Our results indicate that stress can alter lidocaine levels in plasma and tissues, suggesting that stress should be considered an important factor when determining the dosage of lidocaine in clinical application. PMID- 12587952 TI - Comparison of in vitro hepatic models for the prediction of metabolic interaction between simvastatin and naringenin. AB - The study of potential interaction at a very early stage of drug development requires suitable in vitro models that describe drug interactions both qualitatively and quantitatively. The purpose of this work was to help assessing the predictive value of in vitro drug interaction test with liver microsomal fractions and hepatocytes by determination of enzymatic parameters such as the inhibition constant (Ki) and the intrinsic clearance (Clint). This study was conducted to compare different methods of Ki calculation and to determine the most suitable parameter for describing drug interactions. The metabolic interaction between SV and NRG was used as a model to help verifying the suitability of the in vitro model for predicting the kind and degree of metabolic drug interactions. The method of Ki calculation using linearized versions of Michaelis Menten equations based on the simultaneous non linear regressions and the "km app" approach accurately estimated the Ki values. The linear representation of an inherently non linear relationship was only used to establish the mechanism of inhibition and the Clint could be used to predict drug interactions. To further prediction in humans, it seems likely that the simultaneous application of both systems, microsomal fractions and hepatocytes will yield conclusions. PMID- 12587953 TI - Comparison of MEGX (monoethylglycinexylidide) and antipyrine tests in patients with liver cirrhosis. AB - The aim of the study was to compare the feasibility of the MEGX (monoethylglycinexylidide) and antipyrine tests in reference to standard biochemical parameters used for liver assessment in cirrhotic patients. The study was carried out in 44 subjects: 14 healthy controls and 30 cirrhotic patients classified according to the Child-Pugh's score to subgroups A (n=11), B (n=12) and C (n=7). All subjects underwent two dynamic liver tests, i.e. MEGX (monoethylglycinexylidide) and antipyrine test in a crossover schedule with at least 5-day interval. For the MEGX, lidocaine was administrated intravenously, at a dose of 1 mg/kg, and blood samples for MEGX assay were collected after 15 minutes. MEGX concentrations were measured by fluorescence polarization immunoassay. The antipyrine concentrations were evaluated following a single oral administration of 1000 mg antipyrine. The blood was sampled for 24 hours after the drug administration, and antipyrine concentrations were measured spectrophotometrically. Standard biochemical parameters used for liver assessment were measured by means of routine laboratory methods. It was concluded that in patients liver with cirrhosis, liver dynamic tests were better predictors of hepatic function. The MEGX test was more feasible in clinical setting, but it was noted that antipyrine test was more sensitive in staging liver cirrhosis. PMID- 12587955 TI - Calculation of steady-state distribution delay between central and peripheral compartments in two-compartment models with infusion regimen. AB - A lag time may exist between blood drug concentration and drug effect. Various factors can contribute to the lag time, among which the drug distribution delay is a significant one. The drug distribution delay can also exist between different compartments. An equation was derived to calculate the steady-state drug concentration delay between central and peripheral compartments in a two compartment model with infusion regimen. PMID- 12587954 TI - Absolute bioavailability of [14C] genistein in the rat; plasma pharmacokinetics of parent compound, genistein glucuronide and total radioactivity. AB - The systemic plasma pharmacokinetics of genistein were determined in rats to evaluate the absolute oral bioavailability and make comparison with similar data in the literature derived from humans subjects. The plasma concentrations of genistein, genistein glucuronide and carbon-14 were determined by LC-MS/MS and liquid scintillation counting following oral and intravenous dosing with [14C]genistein (4 mg kg(-1) body weight). The absorption of total radioactivity from the gut, (parent compound and metabolites), was 56 and 111% in male and female rats, respectively. In contrast, the absolute oral bioavailability of genistein in male and female rats was 7 and 15%. There was a significant (P<0.001) difference between Cmax of genistein after intravenous (6921 and 4392 ng/ml) and oral (21 and 22 ng/ml) dosing in male and female rats, respectively. After oral administration, the concentration profile of genistein glucuronide in plasma greatly exceeded that of parent compound during the absorption/distribution phase suggesting extensive first pass metabolism, and provided evidence of entero-hepatic circulation. Selective plasma analysis by LC MS/MS, without prior enzymatic hydrolysis, enabled ready discrimination between parent and conjugated metabolites and prevented gross overestimation of genistein bioavailability. Pharmacokinetic parameters Cmax, Tmax and AUC were similar to those reported in humans, which supports the use of the rat model for genistein toxicity studies. PMID- 12587956 TI - Tissue pharmacokinetics of florfenicol in pigs experimentally infected with Actinobacillus pleuropneumoniae. AB - The aim of this study has been to determine the tissue pharmacokinetic parameters of florfenicol in the pigs experimentally infected with Actinobacillus pleuropneumoniae. 21 crossed-bred (Duroc x Landrace x Yorkshire) local species of pigs were infected experimentally with Actinobacillus pleuropneumoniae serotype 1 and confirmed as typical sub-acute pleuropneumonia. A single dose of 20 mg/kg body weight of florfenicol, a novel animal-using antibiotic, was administrated intramuscularly in the pigs and then samples of blood, lung, trachea with bronchi, liver, kidney and muscle were taken at scheduled time points. Drug concentrations were determined by high performance liquid chromatography (HPLC) with an ultraviolet detector via extraction with ethyl acetate under nitrogen flow. The statistic moment theory (SMT) mathematic package was applied to calculate the tissue pharmacokinetic parameters of florfenicol in the infected model. AUC of lung, trachea with bronchi, liver, kidney and muscle were 121.69, 79.37, 81.05, 181.2, and 94.07 mg/l x h, respectively, MRT were from 34.66 to 90.17 h, and t1/2beta from 24.75 to 69.34 h, respectively. CONCLUSIONS: Florfenicol was widely distributed in these tissues and maintained the effective therapeutic concentrations especially in the respiratory tract tissues that are the target organs of Actinobacillus pneuropneumoniae. CLINICAL SIGNIFICANCE: Tissue pharmacokinetic data could be evidence for regime designing of florfenicol in treatment of porcine pleuropneumonia. PMID- 12587957 TI - In vivo evaluation of guargum-based colon-targeted oral drug delivery systems of celecoxib in human volunteers. AB - The present study involved the in vivo evaluation of orally administered guar gum based colon-targeted tablet formulations of celecoxib (colon-targeted tablet-20 or colon-targeted tablet-30) as compared with an immediate release capsule in 15 human volunteers. Blood samples were obtained at different time intervals and the plasma concentration of celecoxib was estimated by reversed phase HPLC. The immediate release capsules of celecoxib might have disintegrated very fast in GI tract and absorbed quickly from stomach and small intestine thereby producing peak plasma concentration (Cmax of 478 +/- 57 ng/ml) within 3.8 +/- 0.1 h (Tmax). Though celecoxib could be seen in plasma after oral administration of colon targeted tablet-20 or colon-targeted tablet-30 between 1 and 2 h, low levels of drug were observed upto 8 h resulting in peak concentration (Cmax) of 78 +/- 6 ng/ml or 88 +/- 15 ng/ml at 10.5 +/- 1.9 h or 13.5 +/- 1.4 h (Tmax) respectively, whereas the immediate release capsules produced peak plasma concentration (Cmax) of 478 +/- 57 ng/ml at 3.8 +/- 0.1 h (Tmax). Colon-targeted tablets showed decreased AUC(0-infinity), Cmax and absorption rate constant, prolonged absorption time (ta), and increased t1/2 in comparison with the immediate release capsules. The results of the study indicated that the guar gum-based colon targeted tablets of celecoxib did not release the drug significantly in stomach and small intestine, but delivered to the colon resulting in a slow absorption of the drug and making it available for local action in the colon. PMID- 12587958 TI - Rapidly distinguishing reversible and irreversible CYP450 inhibitors by using fluorometric kinetic analyses. AB - In this study we have evaluated the reliability of a fluorescence-based method used for rapid identification of irreversible CYP inhibitors (mechanism-based inhibitors). This was accomplished by comparing the time-dependence pattern of IC50 values from fluorometric kinetic measurements. For irreversible CYP inhibitors, IC50 values decreased as incubation proceeded. This was due to progressive inactivation of corresponding enzymes by reactive metabolites generated during the incubation. This change pattern was confirmed using a number of known irreversible CYP inhibitors, including furafylline, midazolam, erythromycin, clarithromycin, oleandomycin, 17alpha-ethynylestradiol and verapamil. The pattern was different in reversible inhibition, depending upon the compounds tested in the fluorometric kinetic assay. For some compounds, such as clotrimazole, IC50 values remained relatively stable, whereas other compounds, such as miconazole, terfenadine and ketoconazole showed a significant increase with incubation time. Monitoring tested compounds by LC-MS/MS during the incubation confirmed that increases of IC50 were probably caused by the loss of inhibitors, resulting from either metabolic degradation, or non-specific binding to microsomal proteins. PMID- 12587959 TI - Is asthma over- or under-diagnosed in athletes? AB - A high prevalence of asthma has been reported in athletes. However, studies in this population usually show an even higher prevalence of airway hyperresponsiveness (AHR) and exercise-induced bronchoconstriction (EIB). This report compares studies on self-reported or physician-diagnosed asthma in athletes with those using objective measures of airflow limitation or airway responsiveness. The higher prevalence of AHR (or EIB) measured in athletes, when compared with the prevalence of self-reported or physician-diagnosed asthma, suggests that abnormal airway responses are common in athletes, although they are infrequently associated with troublesome respiratory symptoms. This may indicate underdiagnosis of asthma in athletes, possibly due to an underreporting of respiratory symptoms or a reduction in perception of nociceptive sensations with repeated exercise over time, or it may simply mean that high-level training is associated with asymptomatic AHR. In athletes, as in the general population, the use of subjective methods such as surveys and questionnaires results in an underestimation ofthe prevalence of airway dysfunction when compared with objective measurements. The significance of these observations is unknown, and there is a need to determine their long-term consequences for athletes. PMID- 12587960 TI - Not 15 but 50% of smokers develop COPD?--Report from the Obstructive Lung Disease in Northern Sweden Studies. AB - BACKGROUND: The prevalence of chronic obstructive pulmonary disease (COPD) according to guidelines of today seems considerably higher than has been reported also in recent literature. AIM: To estimate the prevalence of COPD as defined by British Thoracic Society (BTS) criteria and the recent global initiative for chronic obstructive lung disease (GOLD) criteria. Further aims were to assess the proportion of underdiagnosis and of symptoms in subjects with COPD, and to study risk factors for COPD. METHODS: In 1996, 5892 of the Obstructive Lung Disease in Northern Sweden (OLIN) Study's first cohort could be traced to a third follow-up survey, and 5189 completed responses (88%) were received corresponding to 79% of the original cohort from December 1985. Of the responders, a random sample of 1500 subjects were invited to a structured interview and a lung function test, and 1237 of the invited completed a lung function test with acceptable quality. RESULTS: In ages >45 years, the prevalence of COPD according to the BTS guidelines was 8%, while it was 14% according to the GOLD criteria. The absolutely dominating risk factors were increasing age and smoking, and approximately a half of elderly smokers fulfilled the criteria for COPD according to both the BTS and the GOLD criteria. Family history of obstructive airway disease was also a risk factor, while gender was not. Of those fulfilling the BTS criteria for COPD, 94% were symptomatics, 69% had chronic productive cough, but only 31% had prior to the study been diagnosed as having either chronic bronchitis, emphysema, or COPD. The corresponding figures for COPD according GOLD were 88, 51, and 18%. CONCLUSIONS: In ages >45 years, the prevalence of COPD according to the BTS guidelines was 8%, and it was 14% according to the GOLD criteria. Fifty percent of elderly smokers had developed COPD. The large majority of subjects having COPD were symptomatic, while the proportion of those diagnosed as having COPD or similar diagnoses was small. PMID- 12587961 TI - Omalizumab (Xolair) improves quality of life in adult patients with allergic asthma: a review. AB - Physicians are increasingly aware that asthma causes significant impairment of the patients physical, psychological, and social well-being. Whilst standard clinical endpoints provide significant information on airway status during treatment, it is important to determine whether such improvements overcome the functional impairment that patients have to deal with on a daily basis. As such, assessment of health-related quality of life (QoL) is an important aspect of asthma management in clinical practice. Omalizumab (Xolair) is a recombinant humanized monoclonal anti-immunoglobulin E (IgE) antibody that represents a new therapeutic approach to IgE-mediated diseases such as allergic asthma. Clinical studies show that omalizumab improves the control of allergic asthma whilst reducing steroid consumption, and enhances long-term disease control in patients with recurrent symptoms. Using established and validated QoL methodology, two placebo-controlled clinical studies in adults with moderate-to-severe allergic asthma have shown that patients treated with omalizumab experience a clinically relevant improvement in all aspects of their asthma-related QoL, changes that were significantly superior to those observed for placebo. Such improvements were apparent when omalizumab was added to existing therapy with inhaled corticosteroids, and maintained during a subsequent steroid-reduction phase. Through effective disease control, omalizumab therefore leads to significant improvements in health-related QoL that are meaningful to patients with allergic asthma. PMID- 12587962 TI - Hyperuricemia as a prognostic factor in pulmonary arterial hypertension. AB - Pulmonary arterial hypertension is a malignant disease with a median survival of 3 years. Uric acid levels are elevated in severe heart failure and in states of hypoxemia. Early data suggest a correlation between hyperuricemia and severe pulmonary arterial hypertension. We studied 29 patients with pulmonary arterial hypertension diagnosed and treated between 1998 and 2001. Clinical characteristics (6 min walk test and New York Heart Association class) and hemodynamic parameters (pulmonary artery pressure, pulmonary vascular resistance and cardiac output) were evaluated and correlated to uric acid level in a retrospective study. Uric acid levels correlated positively with New York Heart Association class (r=0.66, P<0.001) and negatively with 6 min walk test (r=-0.35, P=0.03). Uric acid levels were higher in patients who died than in patients who survived at the end ofthe follow-up period (8.8 vs. 5.7 mg/dl, P=0.001). This study shows that uric acid levels are elevated in severe pulmonary arterial hypertension and can be used as a prognostic marker of disease severity. PMID- 12587963 TI - Dietary vitamin C intake is inversely related to cough and wheeze in young smokers. AB - We aimed to investigate whether dietary vitamin C intake, an important antioxidant, is inversely related to self-reported respiratory symptoms in young adults of a community. A random sample of 4300 subjects, aged 20-44 years, living in Bergen, Norway, received a postal questionnaire on respiratory symptoms; 80% responded. Vitamin C intake (mg per week) was estimated from a food-frequency questionnaire asking how often the subject, during the last year, had consumed units of orange juice, oranges, potatoes, carrots and tomatoes. Significant differences in the intake of vitamin C were observed across smoking categories with current smokers having the lowest intake, while there was no variation by gender, age or occupational dust exposure. Dietary vitamin C intake was in univariate analyses inversely related to "morning cough", "chronic cough", "wheeze" and "wheeze ever". After adjusting for gender, age, body mass index, "occupational exposure" pack-years as well as having and stratified on smoking habits in multiple logistic regression analyses, the relationship between dietary vitamin C intake and "cough" and "wheeze" tended to be associated to smoking. The odds ratio (OR) for "morning cough" was 0.68 (95% CI: 0.35-0.95), "chronic cough" OR 0.69 (95% CI: 0.47-1.04) and "wheeze ever" OR 0.75 (95% CI: 0.56-1.01) in current-smokers with dietary vitamin C intake in the upper (> or =395 mg/ week) vs. the lower (<209 mg/week) tertile. The OR for "wheeze" was 0.56 (95% CI: 0.35 0.88) in ex-smokers. The magnitude ofthese effects remained after excluding subjects with supplementary vitamin C intake (n=199) from the statistical analyses. Among young Norwegian adults, having a low prevalence of asthma and high prevalences of smoking-related respiratory symptoms, dietary vitamin C intake may act as an antioxidant and thereby reduce cough and wheeze in smokers having high oxidant stress. PMID- 12587964 TI - Impact of comorbidities on the duration of COPD patients' hospital episodes. AB - The duration of inpatient episodes due to COPD and the factors that affect it have recently been an object of increasing attention, as the aim has been to shorten inpatient periods and thereby to cut health-care costs. All hospital episodes of patients aged over 45 for a primary diagnosis of COPD equal or less than 150 days in duration were drawn from the treatment register maintained by the National Research and Development Centre for Welfare and Health. The lengths of these 152569 inpatient periods were analysed for sex, age and secondary diagnoses by covariance analysis. The mean age of men at the beginning of the hospital episode was 70.6 years and that of women 70.1 years. Men accounted for 76.9% of all inpatient episodes. Covariance analysis ofthe data with age standardised as 70.5 years yielded a mean hospital episode length of 8.9 (95% confidence interval (CI) 8.8-9.0) days. The mean length of hospital episodes without a secondary diagnosis was 7.7 (95% CI 7.6-7.7) days and that with a secondary diagnosis was 10.5 (95% CI 10.5-10.6) days. The longest inpatient episodes were recorded for the patients with secondary diagnoses of pneumonia, 14.7 (95% CI 14.2-15.2) days, and cerebral ischaemia, 14.2 (95% CI 13.5-14.9) days. Concurrent diseases prolonged the hospital episodes of COPD patients. At the beginning of a hospital episode, it is possible to estimate its duration and the need for different treatments based on the patients age and secondary diagnoses. PMID- 12587965 TI - Peroxiredoxin VI in human respiratory system. AB - Peroxiredoxins (Prxs) constitute a novel family of antioxidant proteins, which specifically prevent enzymes from metal-catalyzed oxidation. The localization of a member of the mono-cystein subfamily of Prxs, Prx VI in human respiratory system and its antioxidant properties were investigated. By immunoblotting, the Prx VI was found to be present in human respiratory epithelium. Immunostaining with rabbit polyclonal antibody raised against the Prx VI revealed that the said protein was present in apical areas and mucus of all respiratory airways from trachea to bronchioles. Immunodepletion of the Prx VI profoundly decreased the antioxidant activity of the respiratory epithelium extract. PMID- 12587966 TI - No impairment of peripheral deposition in novel asthmatics treated with an MDI corticosteroid with spacer. AB - Pulmonary distribution and lung functions were evaluated during a 4-month inhaled corticosteroid treatment period in 10 steroid-naive novel asthmatics with normal or slightly reduced lung functions. Patients were given a total daily dose of 1000 microg of beclomethasone dipropionate aerosol twice a day via a pressured metered dose inhaler with a large-volume chamber device (Volumatic, GlaxoSmith Kline, U.K.). Gamma lung scintigraphy and lung function tests were performed before and after 2 months and 4 months. Inhaled 99mTc-labelled beclomethasone dipropionate liposomes were used to assess lung deposition patterns during inhaled steroid therapy. Serum eosinophil cationic protein (ECP) concentration was used as a surrogate marker of asthmatic inflammation. Following beclomethasone treatment, all lung functions were enhanced, but only FVC values showed significant improvement. The FEV1/FVC ratio remained slightly reduced in spite of inhaled corticosteroid therapy. However, the association between changes in improved FVC values and reduced ECP levels proved to be statistically significant. In lung scintigraphy, no evidence of changes in pulmonary deposition patterns were seen during the follow-up period. We conclude that inhaled corticosteroid therapy can lead to improvements in lung functions and surrogate markers of airway inflammation in novel asthma without affecting the peripheral deposition pattern of aerosols. PMID- 12587967 TI - Effects of PEEP on inspiratory and expiratory mechanics in adult respiratory distress syndrome. AB - The purpose of the present study was to assess the mechanical behavior of the respiratory system separately during inspiration and expiration in adult respiratory distress syndrome (ARDS) and the influence of PEEP on any phasic variations ofthe mechanical respiratory parameters. Airways pressure (P), flow (V), and volume (V) signals were recorded in nine patients with ARDS and 10 patients without known respiratory disorder (control group). All patients were artificially ventilated at three levels of positive end-expiratory pressure (PEEP): 0, 5, and 10 hPa. Data were analyzed separately for inspiratory and expiratory records using multiple linear regression analysis (MLRA) according to the equation: P=Ers V+Rrs V'+P0, where Ers and Rrs represent, respectively, the intubated respiratory system elastance and resistance, and P0 the end-expiratory pressure. In the ARDS group expiratory Ers (ErsEXP=45.58 +/- 4.24 hPa/L) was substantially higher (p<0.01) than inspiratory Ers (ErsINSP=36.76 +/- 2.55) with a marked effect of applied PEEP in diminishing the difference between ErsEXP and ErsINSP (p<0.01). For the ARDS group inspiratory Rrs (RrsINSP) decreased significantly with increasing PEEP (PEEP=0: RrsINSP=16.43, PEEP=10: RrsINSP=13.28, p<0.01). The found differences between ErsEXP and ErsINSP could be attributable to an influence of mechanical ventilation by positive airway pressure on pulmonary edemaand interstitial fluid during the inspiratory phase of the respiratory cycle. PMID- 12587968 TI - Tuberculosis and HIV seroprevalence in Lambeth, Southwark and Lewisham, an area of South London. AB - Since the mid-1980s the number of cases of TB notified within the U.K. has continued to rise although the contribution of HIV to this rise remains unclear. A 12-month prospective cohort study was conducted at chest and HIV clinics in four hospitals in Lambeth, Southwark and Lewisham (LSL), an area of South London, to determine the proportion of patients with culture-proven TB infected with HIV. Secondary aims were to determine the proportion of patients with TB and undiagnosed HIV at first presentation to chest clinics, to determine the proportion of patients presenting with TB as an AIDS defining illness (ADI) and to identify risk factors for co-infection with TB and HIV. In chest clinics, demographic data and left-over blood from patients aged 16 or over with culture proven TB was collected, anonymised and HIV tested. In HIV clinics, demographic data on patients with TB already known to be HIV seropositive were also obtained. Twenty-one patients (13%, 95% CI-8-19%) of 159 with culture-proven TB were infected with HIV Four (3%) of 133 patients at first presentation to chest clinics had undiagnosed HIV; two were subsequently diagnosed. Of the 21 patients withTB and HIV, nine (43%) presented with TB as an ADI. Patients with TB and HIV were significantly more likely to be aged between 35 and 55 years compared to HIV seronegative patients [12/21 (57%) vs. 38/138 (28%), P=0.006]. None of the patients from the Indian Subcontinent were HIV seropositive [0/21 vs. 25/138 (18%), P=0.047]. At the present time, universal HIV testing of patients with culture-provenTB in chest clinics within the U.K. is unlikely to significantly reduce the number of patients with undiagnosed HIV. PMID- 12587969 TI - Longitudinal trends in exercise capacity and health status after pulmonary rehabilitation in patients with COPD. AB - Pulmonary rehabilitation (PR) programmes produce initial improvements in exercise tolerance and health status in patients with chronic obstructive pulmonary disease (COPD). However, there is limited data on the longer term effects of PR. This study has examined whether the initial benefits gained in exercise tolerance and health status may be maintained after a 1-year follow-up programme. Sixty-six patients with COPD were assessed with the MRC Dyspnoea Scale and found to be moderately disabled due to dyspnoea (MRC Grades 3 and 4). These patients were then randomised to an 8 week outpatient programme of either exercise training and education (Exercise group) or to education alone (Control group). Exercise performance was assessed with the shuttle walking test and health status assessed with two disease-specific measures, the St Georges Respiratory Questionnaire and the Chronic Respiratory Disease Questionnaire. After PR, all patients were invited to attend monthly follow-up sessions for 1 year. Fifty-six patients were available for follow-up immediately after the programme and were assessed at 6 months and 1 year. This study has shown that the patients in the Exercise group maintained improvements in exercise capacity and health status up to 6 months after an 8 week programme. At 1 year there was a significant difference between the Exercise and Control groups in terms of exercise tolerance due to a considerable decline experienced by the Control group. However, neither group had maintained improvements in health status at 1 year. Further study is required to assess whether benefit may be sustained for a longer period using alternative follow-up strategies. PMID- 12587970 TI - Is inhalation rate important for a dry powder inhaler? Using the In-Check Dial to identify these rates. AB - The fraction of the emitted dose from an inhaler that has the potential to be deposited into the lungs is known as the fine particle dose (also the respirable dose). During inhalation all dry powder inhalers require a 'force' to be created inside the device so that a fine particle dose is generated from the formulation in the metering chamber. This 'force' is formed by the inhalation rate used together with the resistance (and hence design) inside an inhaler. Studies have shown that the fine particle dose is related to the clinical effect whilst other studies have reported that this dose can be dependent on the inhalation rate used. The inhalation technique recommended by the manufacturer of an inhaled device should, therefore, be used. For those dry powder inhalers that demonstrate significant flow dependent dosage emission it is important that patients use the most desirable rate that has been reported. The In-Check Dial is a simple and ease to use meter that can be used to measure the inhalation rate of a patient when they use each of the commonly prescribed inhalers that are currently available. This meter can be used to identify the most suitable inhaler for each individual. PMID- 12587971 TI - Mesenchymal cystic hamartoma of the lung in Cowden's disease. PMID- 12587972 TI - Video-assisted thoracoscopic thyroidectomy for obstructive sleep apnoea. PMID- 12587973 TI - Inhibitors of apoptosis proteins in injury and disease. AB - The inhibitor of apoptosis (IAP) gene family prevents cell death by binding to and inhibiting caspases. In addition to negatively regulating apoptosis, IAPs bind to signaling intermediates and receptors, and participate in diverse cellular functions. Here, we describe the physiological functions of IAPs and their participation in injury and disease processes. PMID- 12587974 TI - The energetics of ion distribution: the origin of the resting electric potential of cells. AB - The relation between the energies of ion movement and ATP hydrolysis is unknown in tissues with widely varying electric potentials. Consequently, we measured the concentration of the nine major inorganic ions in the extra- and intracellular phases in heart, liver, and red cells with resting electrical potentials, E(N), of -86, -28, and -6 mV, respectively, under six different physiological conditions. We calculated the Nernst electric potential and the energy of ion movement between the phases. We found that the energy of ATP hydrolysis was essentially constant, between -54 and -58 kJ/mol, in all tissues and conditions. In contrast, as E(N) decreased, the energies of the Na+ and K+ gradients decreased, with slopes approximating their valence. The difference between the energies of Na+ and K+ gradients remained constant at 17 kJ/mol, which is approximately one third of the energy of ATP hydrolysis, demonstrating near equilibrium of the Na+/K+ ATPase in all tissues under all conditions. All cations, except K+, were pumped out of cells and all anions, except Cl- in liver and red cell, were pumped into cells. We conclude that the energy of ATP was expressed in Na+/K+ ATPase and its linked inorganic ion transporters to create a Gibbs-Donnan near-equilibrium system, an inherent part of which was the electric potential. PMID- 12587975 TI - Novel mechanisms for feedback regulation of phospholipase C-beta activity. AB - The receptor-regulated phospholipase C-beta (PLC-beta) signaling pathway is an important component in a network of signaling cascades that regulate cell function. PLC-beta signaling has been implicated in the regulation of cardiovascular function and neuronal plasticity. The Gq family of G proteins mediate receptor stimulation of PLC-beta activity at the plasma membrane. Mitogens stimulate the activity of a nuclear pool of PLC-beta. Stimulation of PLC beta activity results in the rapid hydrolysis of phosphatidylinositol-4,5 bisphosphate, with production of inositol-1,4,5-trisphosphate and diacylglycerol, intracellular mediators that increase intracellular Ca2+ levels and activate protein kinase C activity, respectively. Diacylglycerol kinase converts diacylglycerol to phosphatidic acid, a newly emerging intracellular mediator of hormone action that targets a number of signaling proteins. Activation of the Gq linked PLC-beta signaling pathway can also generate additional signaling lipids, including phosphatidylinositol-3-phosphate and phosphatidylinositol-3,4,5 trisphosphate, which regulate the activity and/or localization of a number of proteins. Novel feedback mechanisms, directed at the level of Gq and PLC-beta, have been identified. PLC-beta and regulators of G protein signaling (RGS) function as GTPase-activating proteins on Gq to control the amplitude and duration of stimulation. Protein kinases phosphorylate and regulate the activation of specific PLC-beta isoforms. Phosphatidic acid regulates PLC-beta1 activity and stimulation of PLC-beta1 activity by G proteins. These feedback mechanisms coordinate receptor signaling and cell activation. Feedback mechanisms constitute possible targets for pharmacological intervention in the treatment of disease. PMID- 12587976 TI - The overexpression catalase reduces NO-mediated inhibition of endothelial NO synthase. AB - Previously, we have demonstrated that increased superoxide generation plays a role in the nitric oxide (NO)-mediated inhibition of endothelial NO synthase (eNOS) in endothelial cells (ECs) and that the overexpression of SOD1 could reduce the inhibitory effect of NO. However, SOD1 overexpression did not completely abolish the inhibition of eNOS by NO, indicating the presence of other inhibitory mechanisms. Because superoxide can be dismutated into hydrogen peroxide (H2O2), in this study we determined whether exposure of ECs to NO resulted in increased generation of H2O2 and the potential role of H2O2 in eNOS inhibition. Our results indicated that H2O2 levels were increased in response to NO. Using adenoviral-mediated infection, we demonstrated that catalase overexpression both increased basal eNOS activity in the absence of NO and provided a significant protective effect on eNOS activity in the presence of NO. This protective effect was associated with a significant decrease in H2O2 levels in the presence of NO. In conclusion, our results indicate that increased levels of H2O2 may be involved in the inhibition of eNOS by NO and that the scavenging of H2O2 may be useful to prevent eNOS inhibition during treatments that involve inhaled NO or NO donors. PMID- 12587977 TI - Nitric oxide as a modulator of gastric mucin synthesis: role of ERK and p38 mitogen-activated protein kinase activation. AB - Nitric oxide (NO) is an important biological messenger in the regulation of tissue homeostasis and pathophysiological processes. Here, we investigated the effect of NO on gastric mucus glycoprotein (mucin) synthesis, apoptotic processes, and the involvement of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK). Exposure of gastric mucosal cells to NO donor led to a dose-dependent decrease (up to 48%) in mucin synthesis, accompanied by a marked increase in caspase-3 activity and apoptosis. Inhibition of ERK with PD98059 accelerated (up to 23.8%) the NO-induced decrease in mucin synthesis, and cause further enhancement in caspase-3 activity and apoptosis. Blockade of p38 kinase with SB203580 produced reversal in the NO-induced reduction in mucin synthesis, and substantially countered the induced increase in caspase-3 activity and apoptosis. Moreover, caspase-3 inhibitor not only blocked the NO-induced increase in caspase-3 activity but also produced an increase in mucin synthesis. Thus, the detrimental influence of NO on mucin synthesis is closely linked to caspase-3 activation and apoptosis, and involves ERK and p38 kinase participation. Activation of p38 kinase leads to the upregulation of proapoptotic signal, while ERK activation stimulates the anti-apoptotic pathway. PMID- 12587978 TI - Effects of epidermal growth factor and linoleic acid on lipid contents in human intestinal C2BBe1 cells. AB - Epidermal growth factor (EGF) was reported to regulate triacyl glycerol synthesis in various cells. Linoleic acid and its metabolites were thought to modulate the signal transduction of growth factors. This study determined whether linoleic acid regulated the effect of EGF on lipid contents in human intestinal C2BBe1 cells. Confluent cells were incubated with serum-free medium (control), EGF (45 ng/mL), linoleic acid (42 microg/mL), or combined EGF (45 ng/mL) and linoleic acid (42 microg/mL) for 48 h. The results showed EGF and linoleic acid significantly increased intracellular cholesterol and triglyceride levels compared with the control and combined groups. EGF was a more potent stimulator for triacyl glycerol synthesis in C2BBe1 cells than linoleic acid. However, intracellular cholesterol and triglyceride levels did not differ between the control and combined groups. The secretion of cholesterol and triglyceride into the medium by C2BBe1 cells did not differ among four groups. Both EGF and linoleic acid strongly stimulated the expression of EGF receptor mRNA in C2BBe1 cells at 48 h compared with the control and combined groups. Therefore, EGF and linoleic acid increased triacyl glycerol synthesis in C2BBe1 cells through stimulating the expression of EGF receptor mRNA. The effect of EGF and linoleic acid on this lipogenesis was reversed in the presence of both EGF and linoleic acid by downregulating the expression of EGF receptor mRNA. PMID- 12587979 TI - Structural features of proteins responsible for resistance of tryptophan residues to nitrosylation. AB - It is known that potentially reactive groups of the protein molecule may be most efficiently nitros(yl)ated only when located within hydrophobic globules or built into the membrane. N1-nitrosotryptophan (NOW) is a stable product of nitrosation in vitro. However, the NOW fraction in proteins is small in ordinary proteins. It suggests the existence of unknown mechanisms preventing the accumulation of NOW. Here we show that these mechanisms are underlain by the protein structure. Analysis of protein structure databases to explore the atomic surroundings of tryptophan residues revealed preferential selection of certain surroundings. N(E) atoms of tryptophan residues, which are the targets for nitrosation, have usually polar and nucleophilic groups in their environment. Residues of Asp, Glu, Cys, His, and Met act as catalysts of denitrosation (internal denitrosilase). We found that short peptides with the same residues possessed denitrosilase activity even in solution. This selection might explain both the resistance of tryptophan residues in proteins to nitrosation and the mechanisms of chemical communication by means of reversible nitrosation of proteins. PMID- 12587980 TI - p38 kinase and c-Jun N-terminal kinase oppositely regulates tumor necrosis factor alpha-induced vascular cell adhesion molecule-1 expression and cell adhesion in chondrosarcoma cells. AB - We investigated signaling pathways leading to tumor necrosis factor (TNF) alpha induced intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 expression in chondrosarcoma cells, and determined the functional significance of their expression by examining Jurkat T cell adhesion. TNFalpha induced VCAM-1 and ICAM-1 expression and Jurkat T cell binding. Antibody blocking assay indicated that VCAM-1 mediates TNFalpha-induced Jurkat T cell adhesion. TNFalpha caused activation of mitogen-activated protein (MAP) kinase subtypes, extracellular signal-regulated protein kinase, p38 kinase, and c-jun N terminal kinase (JNK). ICAM-1 expression was not altered by the inhibition of MAP kinases. However, VCAM-1 expression and Jurkat T cell adhesion was blocked by the inhibition of p38 kinase, whereas inhibition of JNK enhanced VCAM-1 expression and cell adhesion without any modulation of NFkappaB activation. Our results, therefore, indicate that p38 kinase mediates TNFalpha-induced VCAM-1 expression and cell adhesion, whereas JNK suppresses VCAM-1 expression that is independent to NFkappaB activation. PMID- 12587981 TI - Clonidine displacement from type 1 imidazoline receptor by p-aminobenzamidine, the prototype of trypsin-like serine protease inhibitors. AB - p-Aminobenzamidine inhibits competitively the catalytic activity of enzymes that recognize preferentially the L-arginyl side chain and related structures. Notably, p-aminobenzamidine is considered as the prototype of trypsin-like serine protease inhibitors. Furthermore, p-aminobenzamidine inhibits the catalytic activity of nitric oxide synthase type I and type II as well as copper amine oxidase. Taking into account the structural similarity between p aminobenzamidine, agmatine (the putative endogenous ligand of the membrane type 1 imidazoline receptor (I1-R)), and N-amidino-2-hydroxypyrrolidine (the product of agmatine oxidation by copper amine oxidase), the [3H]clonidine displacement from I1-R in rat heart membranes by p-aminobenzamidine was investigated. p Aminobenzamidine is as effective as agmatine and N-amidino-2-hydroxypyrrolidine and more effective than the antihypertensive drug clonidine to displace [3H]clonidine from I1-R. Therefore, trypsin-like serine protease inhibitors structurally related to p-aminobenzamidine should be administrated under careful control. PMID- 12587982 TI - Q. How can DNA patterns of somatically acquired immunity be imprinted on the germline of immunoglobulin variable (V) genes? PMID- 12587983 TI - How I became a biochemist. PMID- 12587984 TI - Tomatoes and cardiovascular health. AB - Diet is believed to play a complex role in the development of cardiovascular disease, the leading cause of death in the Western world. Tomatoes, the second most produced and consumed vegetable nationwide, are a rich source of lycopene, beta-carotene, folate, potassium, vitamin C, flavonoids, and vitamin E. The processing of tomatoes may significantly affect the bioavailability of these nutrients. Homogenization, heat treatment, and the incorporation of oil in processed tomato products leads to increased lycopene bioavailability, while some of the same processes cause significant loss of other nutrients. Nutrient content is also affected by variety and maturity. Many of these nutrients may function individually, or in concert, to protect lipoproteins and vascular cells from oxidation, the most widely accepted theory for the genesis of atherosclerosis. This hypothesis has been supported by in vitro, limited in vivo, and many epidemiological studies that associate reduced cardiovascular risk with consumption of antioxidant-rich foods. Other cardioprotective functions provided by the nutrients in tomatoes may include the reduction of low-density lipoprotein (LDL) cholesterol, homocysteine, platelet aggregation, and blood pressure. Because tomatoes include several nutrients associated with theoretical or proven effects and are widely consumed year round, they may be considered a valuable component of a cardioprotective diet. PMID- 12587985 TI - Nutritional responses to the presence of inulin and oligofructose in the diets of domesticated animals: a review. AB - Inulin and oligofructose are prebiotic oligosaccharides fermented in the large intestine. This article provides an extensive review of the effects of these oligosaccharides on gastrointestinal characteristics (microflora, pathogen control, epithelial cell proliferation, putrefactive compound production, fecal characteristics, and nutrient digestibility) and systemic metabolism of carbohydrates, nitrogen, lipids, and minerals in dogs, cats, horses, calves, pigs, poultry, and rabbits. In addition, intake of inulin and oligofructose and considerations in their supplementation to animal diets are discussed. Growth performance and meat production in livestock in response to inulin and oligofructose supplementation are addressed. Finally, the possible substitution of antibiotics with fructans in animal diets and directions for future research are presented. PMID- 12587986 TI - Chitin--the undisputed biomolecule of great potential. AB - Of the truly abundant polysaccharides in Nature, only chitin has yet to find utilization in large quantity. Chitin is the second most abundant natural biopolymer derived from exoskeletons of crustaceans and also from cell walls of fungi and insects. Chitin is a linear beta 1,4-linked polymer of N-acetyl-D glucosamine (GlcNAc), whereas chitosan, a copolymer of GlcNAc (approximately 20%) and glucosamine (GlcN, 80%) residues, is a product derived from de-N-acetylation of chitin in the presence of hot alkali. Chitosan is, in fact, a collective name representing a family of de-N-acetylated chitins deacetylated to different degrees. Both chitin/chitosan and their modified derivatives find extensive applications in medicine, agriculture, food, and non-food industries as well. They have emerged as a new class of physiological materials of highly sophisticated functions. Their application versatility is a great challenge to the scientific community and to industry. All these are the result of their versatile biological activity, excellent biocompatibility, and complete biodegradability in combination with low toxicity. Commercial availability of high-purity forms of chitin/chitosan and the continuous appearance of new types of chitin/chitosan derivatives with more and more useful and specific properties have led to an unlimited R&D efforts on this most versatile amino polysaccharide, chitin to find new applications, which are necessary to realize its full potential. Incidentally, this too has become an environmental priority. No doubt, chitin is surely an undisputed biomolecule of great potential. PMID- 12587987 TI - Tea catechins and polyphenols: health effects, metabolism, and antioxidant functions. AB - Increasing interest in the health benefits of tea has led to the inclusion of tea extracts in dietary supplements and functional foods. However, epidemiologic evidence regarding the effects of tea consumption on cancer and cardiovascular disease risk is conflicting. While tea contains a number of bioactive chemicals, it is particularly rich in catechins, of which epigallocatechin gallate (EGCG) is the most abundant. Catechins and their derivatives are thought to contribute to the beneficial effects ascribed to tea. Tea catechins and polyphenols are effective scavengers of reactive oxygen species in vitro and may also function indirectly as antioxidants through their effects on transcription factors and enzyme activities. The fact that catechins are rapidly and extensively metabolized emphasizes the importance of demonstrating their antioxidant activity in vivo. In humans, modest transient increases in plasma antioxidant capacity have been demonstrated following the consumption of tea and green tea catechins. The effects of tea and green tea catechins on biomarkers of oxidative stress, especially oxidative DNA damage, appear very promising in animal models, but data on biomarkers of in vivo oxidative stress in humans are limited. Larger human studies examining the effects of tea and tea catechin intake on biomarkers of oxidative damage to lipids, proteins, and DNA are needed. PMID- 12587988 TI - How does homeopathy work: are we looking in the right place? PMID- 12587989 TI - Users of homeopaths in Norway in 1998, compared to previous users and GP patients. AB - AIM: Homeopathy is the form of complementary medicine most frequently used in Norway. This study describes complaints and characteristics of patients who visited Norwegian homeopaths in 1998, comparing them with those who visited homeopaths in 1985 and general practice patients. METHODS: We conducted a survey of 1097 patients visiting 80 Norwegian homeopaths in 1998 and compared them with a similar survey in 1985 (1072 consultations) and a 1989 survey of general practice patients (90,458 consultations). RESULTS: One in four of patients visiting homeopaths in 1998 were children between 0 and 9 years of age, compared to one in ten in 1985 and in general practice. Almost half of the patients in 1998 had used prescription drugs provided by a medical doctor the previous month for the same complaints they presented to the homeopath. In 1998 patients sought homeopathy most often because of respiratory and skin complaints. In 1985 the most common reasons were musculo-skeletal and digestive problems. Four of the five commonest reasons for encounter in homeopathic practice in 1998 were also found among the five commonest reasons for general practice consultations. CONCLUSION: Patients currently visiting homeopaths differ in age and to some extent in complaints compared to previous users of homeopathy and general practice patients. PMID- 12587990 TI - Influence of the diluent on the effect of highly diluted histamine on basophil activation. AB - BACKGROUND: In modern pharmaceutical practice, it is common to use purified ethanol and purified water for the preparation of homeopathic dilutions. Hahnemann in 1827 recommended good brandy as a diluent. Brandy contains a lot of accompanying substances in addition to ethanol. PURPOSE OF THE STUDY: The research question was whether different diluents influence the effectiveness of high dilutions, especially above Avogadro's number. We compared two dilution media to investigate the diluent's influence. Within the limitations of the test system, the dilution media were as similar to good brandy as possible and like purified ethanol. Dilutions of histamine were prepared with both media. As test system, we used modified basophil activation in an in vitro cell system. Basophils are activated by anti-immunoglobulin E (anti-IgE). The activation of basophils is inhibited by prior incubation with histamine. The reduction in activation was measured with different dilutions of histamine. The test system used a 3-colour flow cytometric method. The interleukin-3 (IL-3) receptor CD123 was used to identify basophils in the leukocyte mixture. The CD63 surface marker was used for quantification of activated basophils. RESULTS: With higher concentrations of histamine, we observed inhibition on optimally anti-IgE stimulated basophil activation with a clear concentration dependency. With low concentrations of histamine (up to 10(-31)), we also observed inhibition of IgE mediated basophil activation. Differences were observed between the dilution media. CONCLUSION: The preliminary results support the hypothesis that the dilution medium may influence the effects of high dilutions. This could be of importance for homeopathic pharmaceutical practice as well as for ultra-high dilution experiments. The refined basophil test system proved to be highly sensitive and reliable. Further studies are needed. PMID- 12587991 TI - A kinetic approach to caffeine--Coffea cruda interaction. AB - The biological effect of Coffea cruda 30c was investigated in rats pre and post treated with caffeine. The experimental subjects were male Wistar rats. Caffeine was administered intraperitoneally at the beginning of a sleep period. Coffea cruda 30c (0.1 ml) was administered orally, a contemporaneous control group was tested. The Electroencephalogram (EEG) was recorded in the parietal region during the following sleep cycle. The effect was evaluated by three EEG parameters: the spectral power in delta (0.5-2.5 Hz) and slow 0.32-0.48 Hz bands and the slow/delta power ratio. These markers were analyzed vs time for control and homeopathic groups, blind. In the pretreated set, a similar pattern was identified for control and verum groups up to the 4th hour. From the 5th hour on, power in the delta band was statistically higher in the verum. Spectral power in the slow band and power ratio for the verum group was smaller than the control group from the 6th hour on. In the post-treated set, two verum sub-groups were identified: Post v-A: did not exhibit significant differences from control; Post v-B: displayed an opposite tendency than pre-treatment verum. We conclude that Coffea cruda 30c modifies sleep pattern increasing sleep intensity with pre treatment. In a subset of the post-treated animals Coffea 30c appeared to reinforce the effects of caffeine. PMID- 12587992 TI - Mutual information and the homeopathic effect. AB - We explore the feasibility of using mutual information to characterize the homeopathic effect. This quantity measures the information gained about a signal at time (t + tau), from its value at an earlier time t; it quantifies the predictability of data. We illustrate our method with an analysis of the homeopathic effect of Strophanthus hispidus on the cardiac rhythm of healthy human subjects, using data from a previous experiment. Our results allow an intuitively clear rendering and agree with the similitude principle applied to this case. They also show that the solvent has a significant effect on the signal; hence, it does not act as an ideal placebo and we discuss some therapeutic corollaries to this observation. PMID- 12587993 TI - Patient-practitioner-remedy (PPR) entanglement. Part 2: Extending the metaphor for homeopathy using molecular quantum theory. AB - A quantum metaphor developed previously for homeopathy, involving triadic patient practitioner-remedy (PPR) entanglement, is extended by importing concepts used in chemistry to describe the electronic structures of molecules. In particular, the electronic energy states of triangular tri-atomic molecules are used metaphorically to predict that (a) the more a homeopathic medicine is potentised, the deeper the level of cure is likely to be, and (b) the practitioner can be included as a beneficiary of the therapeutic process. The model also predicts that remedy attenuation and degree of PRR interaction could (in the quantum theoretical sense) represent a pair of complementary conjugate variables. PMID- 12587994 TI - Homeopathic treatment in emergency medicine: a case series. AB - Following a multiple-casualty construction disaster in Israel, members of The Center of Integrated Complementary Medicine joined in the emergency activity of the Shaare Zedek Medical Center. They administered homeopathic treatment to injured patients to supplement conventional orthopaedic treatment. This was to our knowledge the first time that complementary medicine had been used officially in conjunction with conventional medicine in an emergency situation. Our objective is to report and summarize the rationale, procedures and outcome of the complementary medicine intervention. Fifteen orthopaedic patients were included. They were treated by homeopathy in two phases starting 24 h post-trauma. All patients initially received Arnica montana 200CH in a single dose. Anxiety was treated with Aconite 200CH in nine patients, Opium 200CH in three, Ignatia 200CH in two and Arsenicum album 200CH in one according to type of anxiety. One day later, most patients reported a lessening of pain, 58% felt improvement, 89% had reduced anxiety, and overall, 61% felt that homeopathic treatment was helpful. In the second phase, 48 h post-trauma, specific complaints were addressed with classical homeopathy. At discharge patients rated the homeopathic treatment successful in 67% of the specific complaints. Several issues relating to the use of homeopathy in emergency medicine and its relation to conventional treatment are discussed. These include compliance, the conduct of rounds, shortage of time and staff, and the procurement of medicines. PMID- 12587995 TI - 20 years ago: British Homoeopathic Journal, January 1983. PMID- 12587996 TI - Clinical outcomes research: contributions to the evidence base for homeopathy. PMID- 12587997 TI - A role for small GTPase RhoA in regulating intracellular membrane traffic of lysosomes in invasive rat hepatoma cells. AB - Small GTPase RhoA regulates signal transduction from receptors in the membrane to a variety of cellular events related to cell morphology, motility, cytoskeletal dynamics, cytokinesis, and tumour progression, but it is unclear how RhoA regulates intracellular membrane dynamics of lysosomes. We showed previously by confocal immunofluorescence microscopy that the transfection of dominant active RhoA in MM1 cells causes the dispersal translocation of lysosomes stained for cathepsin D throughout the cytoplasm. Y-27632, a selective inhibitor of p160ROCK, impeded the cellular redistribution of lysosomes and promoted reclustering of lysosomes toward the perinuclear region. Here we have further investigated whether the acidic lysosomal vesicles dispersed throughout the cytoplasm are applied to the early endosomes in the endocytic pathway, and we demonstrate that the dispersed lysosomes were accessible to endocytosed molecule such as dextran, and their acidity was not changed, as determined by increased accumulation of the acidotropic probe LysoTracker Red. Brefeldin A did not induce the tabulation of these dispersed lysosomes, but it caused early endosomes to form an extensive tubular network. The dispersed lysosomes associated with cathepsin D and LIMPII were not colocalized with early endosomes, and these vesicles were not inaccessible to the endocytosed anti-transferrin receptor antibody. Moreover, wortmannin, an inhibitor of phosphatidylinositol 3-kinase, induced a dramatic change in LIMPII-containing structures in which LIMPII-positive swollen large vacuoles were increased and small punctate structures disappeared in the cytoplasm. These swollen vacuoles were not doubly positive for LIMPII and transferrin receptor, and were not inaccessible to the internalized anti transferrin receptor antibody. Therefore, our novel findings presented in this paper indicate that RhoA activity causes a selective translocation of lysosomes without perturbing the machinery of endocytic pathway. PMID- 12587998 TI - Improvement of supersensitive immunohistochemistry with an autostainer: a simplified catalysed signal amplification system. AB - The ImmunoMax/catalysed signal amplification (CSA) system is a supersensitive method of paraffin immunohistochemistry. It incorporates antigen retrieval, the streptavidin-biotin complex (sABC) method, and the catalysing reporter deposition/catalysing biotinylated tyramide reaction. Strong, non-specific cytoplasmic reaction in the ImmunoMax/CSA is due to endogenous biotin unmasked in the antigen retrieval step. We examined procedures to diminish this non-specific immunoreaction and improved the ImmunoMax/CSA. Antigen retrieval in a hot water bath yielded a smaller endogenous biotin immunoreaction than antigen unmasking in an autoclave. Post-antigen retrieval fixation in buffered 10% formalin solution suppressed the biotin immunoreaction but masked the target antigen, Ki67. Post reaction washing with 0.1% Tween 20 in Tris-HCl buffer at 35 degrees C did not diminish the endogenous biotin immunoreaction. Animal serum also did not suppress the non-specific immunoreactivity of biotin and antibodies. Because endogenous biotin is detected by duplicated biotin-streptavidin reactions in the ImmunoMax/CSA, we replaced the sABC step with a labelled polymer secondary antibody (the EnVision system)--a simplified CSA system--because the sensitivity of the EnVision system was the same as that of the sABC method. The non-specific immunoreaction induced by the EnVision system was masked competitively by blocking protein. By using an antibody against Ki67 antigen that can react only with the nucleus, we were able to evaluate the non-specific cytoplasmic immunoreaction induced by the detection system. We believe that the simplified CSA system will open up the field of supersensitive paraffin immunohistochemistry. PMID- 12587999 TI - The expression of Ras-GTPase activating protein SH3 domain-binding proteins, G3BPs, in human breast cancers. AB - Ras-GTPase activating protein SH3 domain-binding proteins 1 and 2 (G3BP1 and G3BP2) have recently been reported to be encoded by two separate genes on human chromosomes 5 and 4 respectively and have been implicated in Ras signalling, NFkappaB signalling, the ubiquitin proteosome pathway and RNA processing. In addition, G3BP1 has recently been implicated in cancer biology. The transcripts for these genes have been shown to be universally expressed; however, this is not the case for the proteins which appear to be tissue and cell type specific. We report here the expression of G3BP1 and 2 in human breast cancers and present the first data showing that G3BP2 expression is specific in human breast cancer tissue and was over-expressed in 88% of tumours examined (n = 58). PMID- 12588000 TI - Visualization of the uptake of high-density lipoprotein by rat aortic endothelial cells and smooth muscle cells in vitro. AB - High-density lipoproteins (HDL) were conjugated to Fluorescein 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) or colloidal gold for the investigation of ultrastructural aspects of binding and uptake of HDL by cholesterol-loaded cultured endothelial and smooth muscle cells from rat aorta. When cells were incubated for 2 h at 4 degrees C, HDL-DiI and HDL-gold conjugates were seen only on the cell surface. When cells were returned to incubation at 37 degrees C for 5 min, HDL-DiI appeared in the cytoplasm and colocalized with the fluorescent cholesteryl ester tag BODIPY-FL-C12. HDL-gold conjugates appeared in the plasmalemmal invaginations and plasmalemmal vesicles. After incubation for 15 min, most of the HDL-gold conjugates reappeared on the cell surface. After incubation for 30 min, only a few conjugates were observed and they localized in lysosomal-like bodies. Quantitative data indicated that when the cholesterol loaded cells were incubated at 4 degrees C for 2 h, the numbers of HDL-gold associated in clusters on the endothelial cell surface was 1.18 clusters/microm. When cells were returned to incubation at 37 degrees C for 5 min, this value decreased to 0.7, increased again to 1.13 at 15 min, and decreased to 0.29 at 30 min. The numbers of clusters in the plasmalemmal invaginations were 0.06 clusters/microm at 4 degrees C for 2 h, increased to 0.34 at 37 degrees C for 5 min and decreased gradually to 0.19 and 0.04 at 15 and 30 min, respectively. The incidence of clusters in the plasmalemmal vesicles per non-nuclear cytoplasm was 0.01 clusters/microm2 at 4 degrees C for 2 h, increased significantly to 1.08 at 37 degrees C for 5 min, and decreased to 0.43 and 0.14 at 15 and 30 min, respectively. This work supports that the plasmalemmal invaginations and plasmalemmal vesicles are linked to the HDL uptake in cholesterol-loaded aortic endothelial cells and smooth muscle cells. PMID- 12588001 TI - Preproenkephalin-like immunoreactive and calcium-binding proteins-like immunoreactive double-labelled neurons in the spinal trigeminal nucleus caudalis of the rat. AB - Immunofluorescence histochemical double-staining for preproenkephalin (PPE) and calbindin-D28k (CB), calretinin (CR) or parvalbumin (PV) were performed in the spinal trigeminal nucleus caudalis (Vc) of the rat. Neuronal cell bodies exhibiting PPE-like immunoreactivity were present in all laminae of the Vc, with a higher concentration in lamina II. Most of the CB-, CR- and PV-like immunoreactive neurons were located in lamina II, and some of them were also found in laminae I and III of the Vc. Some PPE-like immunoreactive neurons also showed CB-, CR-, or PV-like immunoreactivities. CB/PPE, CR/PPE and PV/PPE double labelled neurons were mainly observed in lamina II. The percentages of CB/PPE double-labelled neurons in the total numbers of the CB- and PPE-like immunoreactive neurons were 3.5-1.5% and 3.3-15.7%, respectively. Of all CR- and PPE-like immunoreactive neurons, 4.7-13.5% and 3.7-14.2% showed both CR- and PPE like immunoreactivities. The ratios of PV/PPE double-labelled neurons in all PV- and PPE-like immunoreactive neurons were 9.7-28.1% and 2.1-8.7%, respectively. The present results indicate that some enkephalinergic neurons in the Vc of the rat also contain calcium-binding proteins. PMID- 12588002 TI - Analysis of binding of mannosides in relation to Langerin (CD207) in Langerhans cells of normal and transformed epithelia. AB - Tandem-repeat C-type lectins (pattern-recognition receptors) with specificity for mannosides are intimately involved in antigen recognition, uptake, routing and presentation in macrophages and dendritic cells. In Langerhans cells, Langerin (CD207), a type-II transmembrane protein with a single C-type carbohydrate recognition domain attached to a heptad repeat in the neck region, which is likely to establish oligomers with an alpha-coiled-coil stalk, has been implicated in endocytosis and the formation of Birbeck granules. The structure of Langerin harbours essential motifs for Ca2+-binding and sugar accommodation. Lectin activity has previously been inferred by diminished antibody binding to cells in the presence of the glycan ligand mannan. In view of the complexity of the C-type lectin/lectin-like network, it is unclear what role Langerin plays for Langerhans cells in binding mannosides. In order to reveal in frozen tissue sections to what extent mannose-binding activity co-localizes with Langerin, we have used a synthetic marker, i.e. a neoglycoprotein carrying mannose maxiclusters, as a histochemical ligand, and computer-assisted fluorescence monitoring in a double-labelling procedure. Mannoside-binding capacity was detected in normal epithelial cells. Double labelling ensured the unambiguous assessment of the binding of the neoglycoprotein in Langerhans cells. Light microscopically, its localization profile resembled the pattern of immunohistochemical detection of Langerin. This result has implications for suggesting rigorous controls in histochemical analysis of this cell type, because binding of kit reagents, i.e. mannose-rich glycoproteins horseradish peroxidase or avidin, to Langerin (or a spatially closely associated lectin) could yield false-positive signals. To show that recognition of carbohydrate ligands in dendritic cells is not restricted to mannose clusters, we have also documented binding of carrier-immobilized histo-blood group A trisaccharide, a ligand of galectin-3, which was not affected by the presence of a blocking antibody to Langerin. Remarkably, access to the carbohydrate recognition domain of Langerin appeared to be impaired in proliferatively active environments (malignancies, hair follicles), indicating presence of an endogenous ligand with high affinity to saturate the C-type lectin under these conditions. PMID- 12588003 TI - In situ localization and in vitro expression of osteoblast/osteocyte factor 45 mRNA during bone cell differentiation. AB - The in situ localization of osteoblast/osteocyte factor 45 (OF45) mRNA during bone formation has been examined in the rat mandible from embryonic day 14 (E14) up to postnatal 90-day-old Wistar rats. Gene expression was also examined during cell culture not only in primary rat osteoblast-like cells but also in two clonal rat osteoblastic cell lines with different stages of differentiation, ROB-C26 (C26) and ROB-C20 (C20) using Northern blot analysis. The C26 cell is a potential osteoblast precursor cell line, whereas the C20 cell is a more differentiated osteoblastic cell line. At E15 osteoblast precursor cells differentiated into a group of osteoblasts, some of which expressed the majority of non-collagenous proteins, whereas no expression of OF45 was observed in these cells. Intercellular matrices surrounded by osteoblasts were mineralized at E16. Subsequently, the number of osteoblasts differentiated from osteoblast precursor cells was increased in association with bone formation. At E17, the first expression of OF45 mRNA was observed only in a minority of mature osteoblasts attached to the bone matrix, but not in the rest of less mature osteoblasts. At E20, concomitant with the appearance of osteocytes, OF45 mRNA expression was observed not only in more differentiated osteoblasts that were encapsulated partly by bone matrix but also in osteocytes. Subsequently, osteocytes increased progressively in number and sustained OF45 mRNA expression in up to 90-day-old rats. Northern blot analysis of the cultured cells with or without dexamethasone treatment revealed that the gene expression of OF45 correlated well with the increased cell differentiation. These results indicate that OF45 mRNA is transiently expressed by mature osteoblasts and subsequently expressed by osteocytes throughout ossification in the skeleton and this protein represents an important marker of the osteocyte phenotype and most likely participates in regulating osteocyte function. PMID- 12588004 TI - HlV/STD risks in young men who have sex with men who do not disclose their sexual orientation--six U.S. cities, 1994-2000. AB - To avoid social isolation, discrimination, or verbal or physical abuse, many men who have sex with men (MSM), especially young and minority MSM, do not disclose their sexual orientation. Young MSM who do not disclose their sexual orientation (nondisclosers) are thought to be at particularly high risk for human immunodeficiency virus (HIV) infection because of low self-esteem, depression, or lack of peer support and prevention services that are available to MSM who are more open about their sexuality (disclosers). However, the risks for HIV infection and other sexually transmitted diseases (STDs) are unknown for nondisclosers. To better understand the prevention needs of young MSM, CDC analyzed data from the Young Men's Survey (YMS) to compare HIV/STD risk differences between nondisclosers and disclosers. This report summarizes the results of that analysis, which indicate that 8% of 637 nondisclosers were infected with HIV compared with 11% of 4,952 disclosers. Among blacks, the prevalence of HJV infection was 14% among 199 nondisclosers compared with 24% among 910 disclosers. PMID- 12588005 TI - Hypothermia-related deaths--Philadelphia, 2001, and United States, 1999. AB - Hypothermia is defined as the unintentional lowering of the deep body (core) temperature below 95.0 degrees F (35.0 degrees C). Hypothermia can be mild (90.0 degrees F-<95.0 degrees F [32.2 degrees C-<35.0 degrees C]), moderate (82.5 degrees F-<90.0 degrees F [28.0 degrees C-<32.2 degrees C]), or severe (<82.5 degrees F [<28.0 degrees C]). Common risk factors for hypothermia include exposure to cold while under the influence of alcohol or drugs, altered mental status, and immersion in cold water. During 1979-1998, approximately 700 persons (range: 420-1,024) died annually in the United States from hypothermia; approximately half of these deaths were attributed to extremely cold weather. This report presents three cases of hypothermia-related deaths in Philadelphia during 2001 as examples of risk factors for hypothermia and summarizes information about hypothermia-related deaths in the United States during 1999. Hypothermia deaths are preventable; by avoiding hypothermia, persons also can prevent other adverse health effects of cold weather. PMID- 12588006 TI - Outbreaks of community-associated methicillin-resistant Staphylococcus aureus skin infections--Los Angeles County, California, 2002-2003. AB - During 2002, the Los Angeles County Department of Health Services (LACDHS) investigated three community outbreaks of skin infections associated with methicillin-resistant Staphylococcus aureus (MRSA). MRSA commonly has occurred in health-care settings; however, recent investigations of community-associated MRSA (CA-MRSA) have identified infection in various settings, including correctional facilities, athletic teams, and others (CDC, unpublished data, 2002). This report describes investigations of CA-MRSA in Los Angeles County. PMID- 12588007 TI - Smallpox Vaccine Adverse Events Monitoring and Response System for the first stage of the smallpox vaccination program. AB - Smallpox vaccination of civilian volunteer health-care workers began on January 24, 2003. As of February 4, a total of 37 states and counties have received shipments of smallpox vaccine, and 18 states and counties have begun smallpox vaccination; no serious adverse events have been reported. To monitor the occurrence of adverse events associated with vaccination, both those expected on the basis of previous experience and possible new unexpected adverse events, CDC and state health departments have established the Smallpox Vaccine Adverse Events Monitoring and Response System. The system also will be used to monitor the effectiveness of contraindication screening, identify new contraindications, and coordinate the distribution of vaccinia immune globulin (VIG) and cidofovir to the civilian population. This notice describes the components of the system, delineates roles and responsibilities, and explains how data from the system will be compiled and communicated. PMID- 12588008 TI - WHO and Russia: the turning point in joint efforts against TB. PMID- 12588009 TI - An exploration of the concept of directly observed treatment (DOT) for tuberculosis patients: from a uniform to a customised approach. AB - Tuberculosis (TB) management has moved from chaotic systems and low patient adherence with treatment regimens to the directly observed therapy, short course (DOTS) strategy, which has been described as a new paradigm of TB control. Directly observed treatment (DOT) is only one component of the full DOTS strategy. DOT versus self-administered treatment (SAT) has been the subject of extensive debate, particularly about what approach improves treatment adherence. This debate has been complicated by different case-holding rates and cure outcomes in different contexts where DOT is in place. The increasing range of DOT applications in different settings, including the choice of provider, place, target population, and the extent to which DOT is part of a wider approach, has not been sufficiently taken into account. However, the concrete reality of DOT is an important determinant of the overall success or failure of the programme, and has implications in terms of equity and accessibility of care during treatment. This article aims to go beyond the frequently polarised debate of DOT versus SAT and document the diversity of ways in which DOT has been implemented internationally. We also aim to raise key issues for further discussion, including 1) viewing DOT as part of a complex and lengthy set of interventions that are context-specific, 2) incorporating an equity approach that discusses individual patients' needs and the relationship between the patient and provider, and 3) the role of incentives and enablers. It is anticipated that this exchange of opinion and experiences from different parts of the world will be useful for those involved in the policy formulation and practice of TB management. PMID- 12588010 TI - Low CD4+ T-cell count and high HIV viral load precede the development of tuberculosis disease in a cohort of HIV-positive ethiopians. AB - SETTING: Prospective cohort study, Ethiopia. OBJECTIVE: To study changes in biological markers of HIV infection progression before and after development of TB disease. DESIGN: A longitudinal study of 804 adult factory workers (95 HIV positive, 709 HIV-negative), who were followed every 6 months for a median of 3.8 years. RESULTS: Overall, the incidence rate of TB was 10/ 222 = 45.1 (95%CI 24.3 83.9) per 1000 person-years of observation (PYO) among HIV-1-positive participants, compared to 14/2,054 = 6.8 (95%CI 4.0-11.5) per 1,000 PYO among HIV 1-negative participants (incidence rate ratio 6.62, 95%CI 2.94-14.9). Among the 10 HIV-positive participants who subsequently developed TB disease, the CD4 count was low (median 201/microl, range 45-419), and viral load high (median 4.97 log copies/ml, range 3.70-5.58), at the routine follow-up visit prior to TB diagnosis. Following TB treatment, plasma viral load remained persistently elevated despite clinical resolution of TB disease, and seven of the 10 patients died within a median time of 8 months. CONCLUSION: In this cohort, HIV-infected Ethiopians who developed TB disease already had low CD4 counts and high viral load prior to the diagnosis of TB. Viral load did not decrease following TB treatment, leading to a poor overall prognosis in these patients. PMID- 12588011 TI - Causes of lower respiratory infection in HIV-infected Ugandan adults who are sputum AFB smear-negative. AB - SETTING: Mulago national referral and teaching hospital. OBJECTIVE: To assess the etiology of lower respiratory infections in HIV-infected Ugandan adults who are sputum acid-fast bacilli (AFB) smear-negative. DESIGN: A descriptive cross sectional study. Participants included consecutive adult patients admitted to medical wards with respiratory symptoms of at least 3 weeks and infiltrates on chest radiograph. Those who were sputum AFB smear-negative and HIV-positive underwent bronchoscopy and bronchoalveolar lavage (BAL). BAL fluid was obtained and stained for AFB and Pneumocystis carinii, and cultured for bacteria and fungi. RESULTS: Of 198 patients screened, 48 were sputum smear-positive for AFB. Sixty-seven were excluded for various reasons, leaving 83 patients who met the inclusion criteria and underwent bronchoscopy: 32 (38.6%) patients had P. carinii infection, 20 (24%) had pulmonary tuberculosis, nine (11%) had pulmonary Kaposi's sarcoma and pyogenic bacteria were identified in seven (8%). No aetiological diagnosis was found in 24 (30%) patients. CONCLUSION: P. carinii and Mycobacterium tuberculosis were the commonest causes of disease among sputum AFB smear-negative, HIV-positive adults presenting to Mulago Hospital. Pulmonary Kaposi's sarcoma and pyogenic bacteria occurred with less frequency. PMID- 12588012 TI - Outcome of tuberculosis treatment in Hamburg: a survey, 1997-2001. AB - SETTING: Federal State of Hamburg, Germany, 1997-2001. OBJECTIVE: To determine risk factors affecting the treatment outcome for tuberculosis according to the WHO/IUATLD classification. DESIGN: Prospective evaluation among patients with culture-confirmed pulmonary disease due to Mycobacterium tuberculosis during the period 1997-1999. RESULTS: Five hundred and eighteen (467 new and 51 re treatment) cases started a course of treatment (average duration 36.1 +/- 15.5 weeks), resulting in cure for 416 (80.3%) and treatment completed for three (0.6%) patients; 449 patients (86.7%) initially received a three-drug regimen. Treatment interruption occurred in 54 (10.4%), and failure in 12 (2.3%) cases; 32 (6.2%) patients died (irrespective of cause). Alcohol dependence appeared to be the strongest risk factor for persistence of disease, followed by homelessness and unemployment. The risk of treatment interruption was six times higher among alcoholics (OR = 6.0), five times higher among drug abusers (OR = 5.2) and three times higher among the homeless (OR = 3.0) than in other patients. CONCLUSION: Although the current treatment management in Hamburg is considered to be effective, a further improvement in the proportion of patients who complete treatment can be achieved by increased public health surveillance of subpopulations with the above-mentioned risk factors. PMID- 12588013 TI - The molecular epidemiology of tuberculosis in western Canada. AB - OBJECTIVE: To define the molecular epidemiology of TB in western Canada, and in particular the risk factors for clustering. MEASUREMENTS: We prospectively identified all positive cultures from newly diagnosed cases of TB diagnosed between February 1995 and January 1997 and carried out restriction fragment length polymorphism (RFLP) testing on all isolates. RESULTS: Of 956 cases identified, 944 fulfilled the entry criteria. The mean age was 49.65 years (+/- 22.33), and 508 (53.6%) were males. Three hundred and three (32.1%) subjects were clustered; this varied from 20.2% of the foreign born, 48.4% of Canadian non Aboriginal and 61.1% of all Aboriginal persons. Younger persons (P = 0.0001), males (P = 0.015), those with pulmonary disease (P < 0.001), living in a shelter in the past year (P < 0.001), drug-susceptible disease (P < 0.036), predisposing factors (P < 0.001), prior contact (P < 0.001), and prior skin test (P < 0.002) were more likely to cluster. Among specific risk factors, HIV infection, injection drug use, alcohol excess, and weight loss were all significant. CONCLUSIONS: In this description of the molecular epidemiology of TB in Western Canada, previous results have been confirmed and extended. These results highlight the importance of identifying specific high risk groups, especially in the context of renewed efforts to target persons for treatment of latent TB infection. PMID- 12588014 TI - The epidemiology of tuberculosis among asylum seekers in The Netherlands: implications for screening. AB - OBJECTIVE: To identify low-risk groups among asylum seekers in the Netherlands that may be excluded from tuberculosis (TB) screening at entry or during follow up. METHODS: A retrospective cohort study of medical records of asylum seekers entering the country between January 1994 and March 1997. RESULTS: Medical records were available for 46,424 of the 96,000 asylum seekers (48%) in this period. One hundred and three pulmonary TB cases were diagnosed at entry (prevalence 222/100,000). Risk factors were age >11 years, history of imprisonment and country of origin at war or with TB incidence >100/100,000. During a mean follow-up period of 10 months, 51 pulmonary TB cases were diagnosed (incidence 134/100,000 person-years). Risk factors were age >11 years, old lesions on entry X-ray, and country of origin whose asylum seekers had a prevalence of TB at entry >200/100,000. CONCLUSIONS: We conclude that 1) those with abnormal X-ray at entry should receive preventive therapy after exclusion of active TB, or undergo intensive follow-up, 2) periodic screening is not indicated for immigrants from countries whose asylum seekers have a low prevalence of pulmonary TB at entry, and 3) children <12 years can be excluded from screening. PMID- 12588015 TI - Exogenous re-infection as a cause of recurrent tuberculosis in a low-incidence area. AB - SETTING: Surveillance data from the National Tuberculosis Register for the period 1993-1997 complemented with DNA fingerprinting results of Mycobacterium tuberculosis isolates. OBJECTIVE: To estimate the proportion of disease attributable to recent re-infection among Dutch tuberculosis patients with reported tuberculosis infection or disease before 1981. DESIGN: Data from 1,547 Dutch patients diagnosed between 1994 and 1997 in the Netherlands were studied. Cases with reported tuberculosis infection or disease before 1981 were attributed to reactivation if their M. tuberculosis isolate was unclustered based on DNA fingerprinting or if they were the first case in a cluster, and to re-infection if they were clustered, but not as the first case. RESULTS: In total, 183 Dutch tuberculosis patients (12%) had reported tuberculosis infection or disease before 1981. Tuberculosis in 29 of these patients (16%) was attributed to recent re infection. CONCLUSION: In this setting with a low tuberculosis incidence, approximately one in six new disease episodes among patients with previous tuberculosis infection or disease may be attributable to recent re-infection. PMID- 12588016 TI - Tuberculosis in Nazilli District Prison, Turkey, 1997-2001. AB - SETTING: Nazilli District Prison, Turkey. OBJECTIVE: To evaluate the tuberculosis situation in a Turkish prison. DESIGN: Data obtained between 1997 and 2001 during the systematic annual tuberculosis screening programme undertaken in Nazilli District Prison were evaluated retrospectively. The screening programme consisted of yearly miniature chest radiography, and clinical and bacteriological evaluation of prisoners with abnormalities. Active tuberculosis cases among prisoners diagnosed passively at any time during the period were also studied. RESULTS: Over the 5-year period, 99.8% of the prisoners were screened. The mean point prevalence of tuberculosis in Nazilli District Prison was 341 per 100,000 prisoners. The total number of active tuberculosis cases was 13; five were symptomatic, of whom three were detected during screening. The remaining eight cases were determined only by screening. Treatment outcomes were treatment completion for six cases and cure for seven cases. CONCLUSION: Because tuberculosis prevalence is high among prisoners, it is necessary to continue screening for tuberculosis in prison in addition to passive case-finding activities. Treatment success is satisfactory in Nazilli District Prison. Screening of prison staff, contact examination and preventive approaches need further investigation. PMID- 12588017 TI - Evaluation of undergraduate training on tuberculosis at Istanbul Medical School. AB - SETTING: Undergraduate training on tuberculosis at Istanbul Medical School. OBJECTIVE: To assess whether tuberculosis-related questions asked in chest medicine examinations comply with the World Health Organization's (WHO) learning objectives for tuberculosis training, and to investigate students' skills in interpreting radiology and smears with respect to tuberculosis. DESIGN: Tuberculosis questions set for 4th year medical students (n = 838) were compared with WHO objectives (n = 23): 122 students were each asked to interpret 10 chest X-rays which had been read by three experts as 'typical tuberculosis lesions', 'normal' or 'non-tuberculosis lesions'; 114 students were also each given five smears which had been interpreted by an expert as 'negative' or 'positive' with respect to acid-fast-bacilli (AFB), and were asked to interpret them. RESULTS: Questions covered only nine of the WHO objectives. Among 117 types of questions, 91 (77.8%) were about clinical-radiological findings, treatment and drugs; 334 (65.1%) of 513 X-rays with typical tuberculosis lesions, 77 (21.1%) of 364 normal X-rays and 87 (25.4%) of 343 X-rays with non-tuberculosis lesions were identified as tuberculosis (kappa 0.45). Of 369 AFB-positive smears, 149 (40.4%) were evaluated as false negative, and of 185 AFB-negative smears, 48 (25.9%) were evaluated as false positive (kappa 0.49). CONCLUSION: Examination questions set on tuberculosis at Istanbul Medical School do not adequately reflect WHO learning objectives. Students' skills in interpreting radiology and smears suggest that their practical training on tuberculosis is insufficient. PMID- 12588018 TI - Private tuberculosis care provision associated with poor treatment outcome: comparative study of a semi-private lung clinic and the NTP in two urban districts in Ho Chi Minh City, Vietnam. National Tuberculosis Programme. AB - SETTING: Ho Chi Minh City, Vietnam. OBJECTIVE: To compare tuberculosis case management and treatment outcome between a semi-private chest clinic and a publicly run national tuberculosis programme (NTP). METHOD: Prospective, non randomised, comparative cohort study. Case-management and treatment outcome was determined for 176 patients treated in the semi-private clinic and 326 patients treated in the NTP. RESULTS: In the semi-private clinic cohort, significantly fewer patients completed treatment and/or were cured than in the NTP cohort (48.9% vs. 85.0%, P < 0.001). Among patients with sputum-positive pulmonary TB, significantly fewer were cured in the semi-private clinic cohort compared to the NTP cohort (22.2% vs. 79.2%, P < 0.001), and treatment success was significantly lower (35.2% vs. 79.7%. P < 0.001). Adjustment for a number of potential confounders did not change these findings significantly. CONCLUSIONS: Treatment outcome was considerably better in the NTP than in the semi-private clinic. The difference is not likely to be due to differences in patient characteristics or in provider knowledge. Different financial incentives for the providers in the two settings and ways of paying for services by patients are possible reasons for the observed difference in the quality of case management and treatment outcome. PMID- 12588019 TI - PPD RT23 for tuberculin surveys in India. AB - SETTING: Tuberculosis sanatoria and villages in Bangalore district. OBJECTIVES: To study the appropriateness of continuing to use 1TU dilutions prepared by the BCG Laboratory, Guindy, in Chennai, India, from a freeze-dried form of PPD RT23 with Tween 80 received from Statens Serum Institut (SSI), Copenhagen, for tuberculin surveys in India. DESIGN: The responses to dual tuberculin tests were compared among: 1) 63 smear-positive cases using 2TU PPD prepared by the Guindy laboratory (Dilution-G), and 2TU PPD prepared by the SSI (Dilution-S); 2) 124 smear-positive cases using 1TU and 2TU Dilution-G; and 3) 1,338 apparently healthy children using 1TU and 2TU Dilution-G. Test sites were allocated randomly using the double-blind technique. Tuberculin responses obtained during studies conducted in India and in other countries were compared. RESULTS: The differences in sensitivity of tuberculin testing using the different preparations were found to be small and statistically non-significant. Among children, a higher proportion of reaction sizes in 10-14 mm and 15 + mm categories was observed to 2TU compared to 1TU of Dilution-G. This could reflect lower specificity of 2TU in the study area where non-specific sensitivity is highly prevalent. Studies in India and other countries do not suggest any loss in potency of 1TU PPD RT23 with Tween 80. CONCLUSION: 1TU dilutions of PPD RT23 with Tween 80 provided by the BCG Laboratory, Guindy, may continue to be used for tuberculin surveys in India. PMID- 12588020 TI - Epidemiology of Mycobacterium bovis disease in San Diego County, 1994-2000. AB - SETTING: Local public health department. OBJECTIVE: To examine the current epidemiology of Mycobacterium bovis disease in San Diego County and to compare patient characteristics to those of patients with M. tuberculosis disease. DESIGN: Retrospective review of surveillance and epidemiologic data. RESULTS: Between 1994 and 2000, 1,931 evaluable cases of culture-positive TB were identified; 129 were infected with M. bovis and 1,802 were infected with M. tuberculosis. More than 90% of M. bovis cases occurred in Hispanic persons. About 25% of patients were children, and the main site of disease was extra-pulmonary in 53% of patients. Twenty-three per cent had concurrent HIV infection. Compared to patients with M. tuberculosis, patients with M. bovis were more likely to be Hispanic, aged 0-14 years, have extra-pulmonary disease, or have HIV co-infection in a multivariate model. They were less likely to be born in countries outside of the US or Mexico. CONCLUSION: While human disease due to M. bovis has essentially been eliminated in many parts of the US and other industrialized countries, it persists in certain demographic groups, including Hispanic children and HIV infected adults, in San Diego County. PMID- 12588021 TI - A comprehensive study of the efficiency of the routine pulmonary tuberculosis diagnostic process in Nairobi. AB - Tuberculosis (TB) suspects from Rhodes Chest Clinic, Nairobi, Kenya, were subjected to three sputum smear microscopy (Ziehl-Neelsen) examinations and a chest X-ray (CXR). Results were compared with Lowenstein-Jensen culture as the gold standard to establish the efficiency of the routine diagnostic process. All laboratory tests and the CXR were available for 993 (71%) of the 1,398 enrolled suspects. Of these, 554 (56%) were culture-positive. The routine diagnostic process was very sensitive, able to detect 92% of culture-positive cases but missing 8%. The specificity was low (66%), and 23% of the patients started on treatment were culture-negative, mainly due to the low specificity of the CXR. It may be possible to increase the efficiency of the diagnostic process by specifying better criteria for CXR examination, improving the quality of CXR reading and counselling patients to return when complaints persist. PMID- 12588022 TI - Mycobacterium tuberculosis complex is not detected by DNA amplification assay in sputum specimens of patients with lung scars due to past pulmonary tuberculosis. AB - OBJECTIVE: To evaluate detection of false-positive sputum amplification assay results in former tuberculosis patients with residual pulmonary scars. DESIGN: A total of 268 sputum specimens from 25 war veterans with tuberculosis during 1940 1959, without adequate chemotherapy, and 19 subjects effectively treated for cavitary tuberculosis during 1980-1993 were tested by smear, culture and DNA amplification, as were 34 controls with no history of tuberculosis or pulmonary scars. RESULTS: No active tuberculosis cases were identified. All specimens were negative on DNA amplification and smear. Eight specimens from six subjects were positive on culture, revealing atypical mycobacteria. CONCLUSION: No genetic Mycobacterium tuberculosis material in sputum specimens of subjects with residual lesions of pulmonary tuberculosis and no false-positive amplification results were detected. PMID- 12588023 TI - Standard errors in mixture studies of tuberculous infection prevalence. AB - Tuberculous infection status in individuals assessed by skin injections of 5 TU PPD-S tuberculin are subject to error. Some truly infected persons may respond with a small skin induration, and some not infected with Mycobacterium tuberculosis can respond with a large induration. Rust and Thomas used data from 701,317 white US Navy recruits seen during 1961-1968 to describe induration size distributions among those infected and not infected, and to provide prevalence estimates. Here, standard errors of prevalence and induration size probabilities are presented through use of the jack-knife methodology. PMID- 12588024 TI - The utility of an antibiotic trial for diagnosis of AFB-negative tuberculosis. PMID- 12588025 TI - Tuberculosis and HIV co-infection in children under 15 years of age in Rio de Janeiro, Brazil. PMID- 12588026 TI - Prediction of residual pleural thickening in tuberculous pleural effusion. PMID- 12588027 TI - DOTS-S: directly observed therapy short course-with smiles. PMID- 12588028 TI - Validation of medical image processing in image-guided therapy. PMID- 12588029 TI - Automatic centerline extraction for virtual colonoscopy. AB - In this paper, we introduce a concise and concrete definition of an accurate colon centerline and provide an efficient automatic means to extract the centerline and its associated branches (caused by a forceful touching of colon and small bowel or a deep fold in twisted colon lumen). We further discuss its applications on fly-through path planning and endoscopic simulation, as well as its potential to solve the challenging touching and colon collapse problems in virtual colonoscopy. Experimental results demonstrated its centeredness, robustness, and efficiency. PMID- 12588030 TI - Edge displacement field-based classification for improved detection of polyps in CT colonography. AB - Colorectal cancer can easily be prevented provided that the precursors to tumors, small colonic polyps, are detected and removed. Currently, the only definitive examination of the colon is fiber-optic colonoscopy, which is invasive and expensive. Computed tomographic colonography (CTC) is potentially a less costly and less invasive alternative to FOC. It would be desirable to have computer aided detection (CAD) algorithms to examine the large amount of data CTC provides. Most current CAD algorithms have high false positive rates at the required sensitivity levels. We developed and evaluated a postprocessing algorithm to decrease the false positive rate of such a CAD method without sacrificing sensitivity. Our method attempts to model the way a radiologist recognizes a polyp while scrolling a cross-sectional plane through three dimensional computed tomography data by classification of the changes in the location of the edges in the two-dimensional plane. We performed a tenfold cross validation study to assess its performance using sensitivity/specificity analysis on data from 48 patients. The mean specificity over all experiments increased from 0.19 (0.35) to 0.47 (0.56) for a sensitivity of 1.00 (0.95). PMID- 12588031 TI - Characterization of tomographic sampling in hybrid PET using the Fourier crosstalk matrix. AB - Hybrid positron emission tomography (PET) cameras can be used to measure the distribution of positron emitting radionuclides. An important system parameter for Hybrid PET is the appropriate tomographic sampling requirements. In this paper, a previously developed theoretical formulation for quantifying sampling in continuous-to-discrete tomographic systems, termed the "crosstalk matrix," is used to provide information on the recoverability of the Fourier coefficients that represent the continuous object. In addition, the crosstalk matrix can be related to image quality assessment. Here, we use the crosstalk matrix to evaluate tomographic sampling for Hybrid PET systems. Dual-and triple-head systems were compared, with emphasis placed on studying how system performance changes as the number of gantry stops is increased, and as the line-of-response acceptance angle is reduced. Examination of the crosstalk matrix, as well as figures-of-merit measuring task performance that are computed using the crosstalk matrix, show that increasing angular sampling improves Fourier coefficient recoverability and reduces aliasing. PMID- 12588032 TI - Feature-guided shape-based image interpolation. AB - A feature-guided image interpolation scheme is presented. It is an effective and improved, shape-based interpolation method used for interpolating image slices in medical applications. The proposed method integrates feature line-segments to guide the shape-based method for better shape interpolation. An automatic method for finding these line segments is given. The proposed feature-guided shape-based method can manage translation, rotation and scaling situations when the slices have similar shapes. It can also interpolate intermediate shapes when the successive slices do not have similar shapes. This method is experimentally evaluated using artificial and real two-dimensional and three-dimensional data. The proposed method generated satisfactory interpolated results in these experiments. We demonstrate the practicality, effectiveness and reproducibility of the proposed method for interpolating medical images. PMID- 12588033 TI - A high-resolution technique for ultrasound harmonic imaging using sparse representations in Gabor frames. AB - Over the last few decades there were dramatic improvements in ultrasound imaging quality with the utilization of harmonic frequencies induced by both tissue and echo-contrast agents. The advantages of harmonic imaging cause rapid penetration of this modality to diverse clinical uses, among which myocardial perfusion determination seems to be the most important application. In order to effectively employ the information, comprised in the higher harmonics of the received signals, this information should be properly extracted. A commonly used method of harmonics separation is linear filtering. One of its main shortcomings is the inverse relationship between the detectability of the contrast agent and the axial resolution. In this paper, a novel, nonlinear technique is proposed for separating the harmonic components, contained in the received radio-frequency images. It is demonstrated that the harmonic separation can be efficiently performed by means of convex optimization. It performs the separation without affecting the image resolution. The procedure is based on the concepts of sparse signal representation in overcomplete signal bases. A special type of the sparse signal representation, that is especially suitable for the problem at hand, is explicitly described. The ability of the novel technique to acquire "un-masked," second (or higher) harmonic images is demonstrated in series of computer and phantom experiments. PMID- 12588034 TI - A knowledge-based approach to automatic detection of the spinal cord in CT images. AB - Accurate planning of radiation therapy entails the definition of treatment volumes and a clear delimitation of normal tissue of which unnecessary exposure should be prevented. The spinal cord is a radiosensitive organ, which should be precisely identified because an overexposure to radiation may lead to undesired complications for the patient such as neuronal disfunction or paralysis. In this paper, a knowledge-based approach to identifying the spinal cord in computed tomography images of the thorax is presented. The approach relies on a knowledge base which consists of a so-called anatomical structures map (ASM) and a task oriented architecture called the plan solver. The ASM contains a frame-like knowledge representation of the macro-anatomy in the human thorax. The plan solver is responsible for determining the position, orientation and size of the structures of interest to radiation therapy. The plan solver relies on a number of image processing operators. Some are so-called atomic (e.g., thresholding and snakes) whereas others are composite. The whole system has been implemented on a standard PC. Experiments performed on the image material from 23 patients show that the approach results in a reliable recognition of the spinal cord (92% accuracy) and the spinal canal (85% accuracy). The lamina is more problematic to locate correctly (accuracy 72%). The position of the outer thorax is always determined correctly. PMID- 12588035 TI - Computerized detection of pulmonary embolism in spiral CT angiography based on volumetric image analysis. AB - A fully automated method for computerized detection of pulmonary embolism in spiral computed tomography angiography was developed based on volumetric image analysis. The detection method is based on segmentation of pulmonary vessels to limit the search space, and analysis of several three-dimensional features inside segmented vessel volume. The features utilized are vascular size, local contrast based on mathematical morphology, degree of curvilinearity based on second derivatives, and geometric features such as volume and length. Detection results were obtained for 19 clinical data sets and the performance of the method was evaluated. Using the number and locations of thrombi diagnosed by radiologists as the gold standard, 100% sensitivity was achieved with 7.7 false positives per case, and 85% sensitivity was obtained with 2.6 false positives. For identification of all the positive cases as positive, i.e., detection of at least one thrombus per positive case, 1.9 false positives per case were obtained. These preliminary results suggest that the method has potential for fully automated detection of pulmonary embolism. PMID- 12588036 TI - Implementation, calibration and accuracy testing of an image-enhanced endoscopy system. AB - This paper presents a new method for image-guided surgery called image-enhanced endoscopy. Registered real and virtual endoscopic images (perspective volume renderings generated from the same view as the endoscope camera using a preoperative image) are displayed simultaneously; when combined with the ability to vary tissue transparency in the virtual images, this provides surgeons with the ability to see beyond visible surfaces and, thus, provides additional exposure during surgery. A mount with four photoreflective spheres is rigidly attached to the endoscope and its position and orientation is tracked using an optical position sensor. Generation of virtual images that are accurately registered to the real endoscopic images requires calibration of the tracked endoscope. The calibration process determines intrinsic parameters (that represent the projection of three-dimensional points onto the two-dimensional endoscope camera imaging plane) and extrinsic parameters (that represent the transformation from the coordinate system of the tracker mount attached to the endoscope to the coordinate system of the endoscope camera), and determines radial lens distortion. The calibration routine is fast, automatic, accurate and reliable, and is insensitive to rotational orientation of the endoscope. The routine automatically detects, localizes, and identifies dots in a video image snapshot of the calibration target grid and determines the calibration parameters from the sets of known physical coordinates and localized image coordinates of the target grid dots. Using nonlinear lens-distortion correction, which can be performed at real-time rates (30 frames per second), the mean projection error is less than 0.5 mm at distances up to 25 mm from the endoscope tip, and less than 1.0 mm up to 45 mm. Experimental measurements and point-based registration error theory show that the tracking error is about 0.5-0.7 mm at the tip of the endoscope and less than 0.9 mm for all points in the field of view of the endoscope camera at a distance of up to 65 mm from the tip. It is probable that much of the projection error is due to endoscope tracking error rather than calibration error. Two examples of clinical applications are presented to illustrate the usefulness of image-enhanced endoscopy. This method is a useful addition to conventional image-guidance systems, which generally show only the position of the tip (and sometimes the orientation) of a surgical instrument or probe on reformatted image slices. PMID- 12588037 TI - Phase-based multidimensional volume registration. AB - We present a method for accurate image registration and motion compensation in multidimensional signals, such as two-dimensional (2-D) X-ray images and three dimensional (3-D) computed tomography/magnetic resonance imaging volumes. The method is based on phase from quadrature filters, which makes it robust to noise and temporal intensity variations. The method is equally applicable to signals of two, three or higher number of dimensions. We use parametric models, e.g., affine models, finite elements or local affine models with global regularization. Experimental results show high accuracy for 2-D and 3-D motion compensation. PMID- 12588038 TI - Cortex segmentation: a fast variational geometric approach. AB - An automatic cortical gray matter segmentation from a three-dimensional (3-D) brain images [magnetic resonance (MR) or computed tomography] is a well known problem in medical image processing. In this paper, we first formulate it as a geometric variational problem for propagation of two coupled bounding surfaces. An efficient numerical scheme is then used to implement the geodesic active surface model. Experimental results of cortex segmentation on real 3-D MR data are provided. PMID- 12588040 TI - A proof of the spherical homeomorphism conjecture for surfaces. AB - The human cerebral cortex is topologically equivalent to a sphere when it is viewed as closed at the brain stem. Due to noise and/or resolution issues, magnetic resonance imaging may see "handles" that need to be eliminated to reflect the true spherical topology. Shattuck and Leahy present an algorithm to correct such an image. The basis for their correction strategy is a conjecture, which they call the spherical homeomorphism conjecture, stating that the boundary between the foreground region and the background region is topologically spherical if certain associated foreground and background multigraphs are both graph-theoretic trees. In this paper, we prove the conjecture, and its converse, under the assumption that the foreground/background boundary is a surface. PMID- 12588039 TI - A support vector machine approach for detection of microcalcifications. AB - In this paper, we investigate an approach based on support vector machines (SVMs) for detection of microcalcification (MC) clusters in digital mammograms, and propose a successive enhancement learning scheme for improved performance. SVM is a machine-learning method, based on the principle of structural risk minimization, which performs well when applied to data outside the training set. We formulate MC detection as a supervised-learning problem and apply SVM to develop the detection algorithm. We use the SVM to detect at each location in the image whether an MC is present or not. We tested the proposed method using a database of 76 clinical mammograms containing 1120 MCs. We use free-response receiver operating characteristic curves to evaluate detection performance, and compare the proposed algorithm with several existing methods. In our experiments, the proposed SVM framework outperformed all the other methods tested. In particular, a sensitivity as high as 94% was achieved by the SVM method at an error rate of one false-positive cluster per image. The ability of SVM to out perform several well-known methods developed for the widely studied problem of MC detection suggests that SVM is a promising technique for object detection in a medical imaging application. PMID- 12588041 TI - High-dose calcium stimulation test in a case of insulinoma masquerading as hysteria. AB - It is reported that some cases with insulinoma present with neuropsychiatric symptoms and are often misdiagnosed as psychosis. Here we report a case of insulinoma masquerading as hysteria, whose final diagnosis could be made using high-dose calcium stimulation test. A 28-yr-old woman was referred presenting with substupor, mutism, mannerism, restlessness, and incoherence. Laboratory examinations revealed hypoglycemia (33 mg/dL) and detectable insulin levels (9.7 microU/mL), suggesting the diagnosis of insulinoma. However, neither imaging studies nor selective arterial calcium injection (SACI) test with a conventional dose of calcium (0.025 mEq/kg) indicated the tumor. High-dose calcium injection (0.05 mEq/kg) evoked insulin secretion when injected into superior mesenteric artery. A solitary tumor in the head of the pancreas was resected, and her plasma glucose returned to normal. Postoperatively, iv injection of secretin resulted in a normal response of insulin, which was not found preoperatively. This case suggests the usefulness of the SACI test with high-dose of calcium in the case of insulinoma when the standard dose fails to detect such a tumor. PMID- 12588042 TI - Blockade of menstrual cycle by thyroidectomy in Japanese monkeys (Macaca fuscata fuscata). AB - To examine the role of thyroid hormones in the seasonal breeding cycle in Japanese monkeys (Macaca fuscata fuscata), sexually mature females were thyroidectomized (n = 6) in early December, during the midbreeding season, or they received sham operations (n = 4). They were housed indoors individually, and blood samples were collected two to three times a week to monitor gonadotropin and gonadal steroid hormone secretions. Control monkeys exhibited ovulatory cycles during the breeding season. The mean dates of onset and end of the ovulatory cycles were October 22 +/- 13 d and February 25 +/- 14 d, respectively. These dates coincided well with those of our colonies under captivity. By contrast, three of the six thyroidectomized monkeys terminated ovulatory cycles immediately after operations; the remaining three monkeys ovulated only once or twice after thyroid removal. The mean dates of onset and end of the ovulatory cycles of thyroidectomized monkeys were October 18 +/- 4 d and December 31 +/- 4 d, respectively. This was a significantly earlier termination of the ovulatory cycles than in controls. Mean concentrations of plasma thyroxine of control monkeys were maintained throughout the experimental period, whereas plasma thyroxine concentrations of thyroidectomized monkeys decreased abruptly to undetectable levels. Thyroidectomized monkeys exhibited significantly higher levels of plasma prolactin (PRL) than controls. Moreover, even in control monkeys, plasma PRL increased during the transition out of the breeding season. These results suggest that thyroid hormones play an important role in the regulation of ovulatory cycles in Japanese monkeys, directory or indirectly, possibly by mediating the changes of PRL secretion. PMID- 12588043 TI - Antiprolactin autoantibodies are associated with hyperprolactinemic status in men infected with human immunodeficiency virus. AB - High serum prolactin (PRL) levels and even hyperprolactinemia are a common finding in human immunodeficiency virus (HIV) infection. However, little is known regarding the mechanisms that may contribute to the rise of PRL. We measured serum PRL levels in 54 HIV-infected and 85 healthy age-matched men. The association between PRL levels among anti-PRL autoantibodies and other clinical variables in HIV-infected men was studied. We also evaluated the changes in serum PRL levels by chromatographic separation (affinity with protein G and gel filtration) after a 10-mg iv bolus of metoclopramide. HIV-infected men had higher serum PRL levels compared with healthy men. Sera from 9 of the 54 (16.7%)HIV infected men were found to have hyperprolactinemia. Moreover, the anti-PRL autoantibody was present in four of nine (44.4%)HIV-infected men with hyperprolactinemia; it was also associated with hyperprolactinemic status. Serum total PRL levels were higher in HIV-infected men with anti-PRL autoantibodies than hyperprolactinemic HIV-infected men without anti-PRL autoantibodies; by contrast, free PRL levels were lower. In HIV-infected men with anti-PRL autoantibodies, gel filtration showed that big big PRL isoform was present as the predominant circulating form of PRL throughout each measurement after iv metoclopramide. By contrast, the predominant isoform of PRL in serum from healthy men and HIV-infected men who were anti-PRL autoantibody negative was little PRL. On the other hand, high serum total PRL levels were observed at each measurement throughout the metoclopramide test in HIV-infected men with anti-PRL autoantibodies; however, the serum free PRL levels were similar to those found in subjects without anti-PRL autoantibodies. These data demonstrated that anti-PRL autoantibodies are associated with hyperprolactinemic status in HIV-infected subjects, particularly in those with high serum PRL levels. PMID- 12588044 TI - Differential responses of an invariant region in the ectodomain of three glycoprotein hormone receptors to mutagenesis and assay conditions. AB - The glycoprotein hormone receptors-luteinizing hormone receptor (LHR), follicle stimulating hormone receptor (FSHR), and thyroid-stimulating hormone receptor (TSHR)--are G-protein-coupled receptors with an invariant 10-amino acid residue sequence in the ectodomain proximal to transmembrane helix 1. A Glu-Asp, located at the midpoint of this conserved sequence, has been suggested to be important in ligand-mediated signaling of LHR and/or receptor expression or stability, but not binding. One goal of this study was to expand the studies on LHR and determine whether the invariant Glu and Asp residues were functional in FSHR and TSHR as well. Another goal was to investigate systematically the role of ionic strength, particularly Na+, which appears to have enigmatic functions in the three receptors regarding ligand binding and receptor activation, and to ascertain whether any of the purported effects of Na+ could involve the conserved pair of acidic side chains in the ectodomain. COS-7 cells were transiently transfected with cDNAs to the wild-type (WT) receptor (rat) and identical single and double mutants of each (Glu --> Ala, Asp; Asp --> Ala, Glu; and Glu-Asp--> Asp-Glu), followed by characterization of cognate ligand binding and signaling (basal and hormone mediated) in two commonly used buffer systems: Waymouth's medium, containing a near-physiologic concentration of Na+ (132 mM); a low ionic strength buffer with a 1 mM concentration of Na+. The three receptors exhibited differential responses to mutagenesis and the two buffers. Notably, a comparison of basal cyclic adenosine monophosphate (cAMP) production showed that the buffer of lower ionic strength resulted in increased basal cAMP production in WT TSHR but not LHR and FSHR; that the maximal ligand-mediated cAMP production was greatest in the buffer of higher ionic strength for the three WT receptors; that functionality of the conserved Glu and Asp residues in ligand-mediated signaling was buffer dependent in LHR, whereas it did not appear to be particularly important in FSHR and TSHR signaling; and that apparent ligand binding in WT and mutant TSHRs seemed to be particularly diminished in the buffer of higher ionic strength. These results demonstrate that identical amino acid residues in homologous receptors can exhibit distinct functions; moreover, the role of ionic strength (Na+) on signaling differs in the three receptors. PMID- 12588045 TI - Intrinsic responses of rat coronary arteries in vitro: influence of testosterone, calcium, and effective transmural pressure. AB - Administration of testosterone to men with angina has been shown to reduce myocardial ischemia. The mechanism of this effect is not clear but could be via an influence on coronary artery tone. We therefore employed an animal model to study the intrinsic responses of coronary arteries mounted in the wire myograph and evaluated the effect of testosterone on coronary artery tone in vitro. Intrinsic responses of the vessels and response to addition of testosterone were observed. Immediately after loading, vessels relaxed, an effect that was dependent on the baseline transmural pressure of vessels and was attenuated by pretreatment with N-nitro-L-arginine methyl ester. Subsequent contraction to a peak wall tension (intrinsic tone) was abolished by removal of extracellular calcium. Addition of testosterone produced a significant dose-dependent relaxation of intrinsic tone in all groups studied. The lowest concentration at which relaxation occurred was 10-6M. We conclude that rat coronary arteries exhibit calcium-dependent intrinsic responses and develop spontaneous tone. Furthermore, addition of testosterone reduces intrinsic coronary artery tone. These findings may have important implications for men with angina and low plasma testosterone levels. PMID- 12588046 TI - Cardiovascular action of insulin-like growth factor-1 is not mediated by calcitonin gene-related peptide neurons. AB - Calcitonin gene-related peptide (CGRP) is a potent vasodilator located in the peripheral nerves including the perivascular nerves. Previous studies in our laboratory determined that the vasodilatory action of insulin is mediated in part by CGRP-containing neurons. Since insulin-like growth factor-1 (IGF-1) and insulin share molecular and receptor structural similarity as well as functional similarity, we investigated the role of the CGRP-containing neurons in IGF-1 mediated vasodilation. Wistar rats were made CGRP deficient by treatment with capsaicin (50 mg/kg) 1-3 d after birth. Vehicle-treated controls and CGRP deficient rats were maintained for 12 to 13 wk. At this time rats were fasted overnight, anesthetized with urethane and chloralose, and prepared for cardiovascular recordings. The basal mean arterial pressure (MAP) was higher in CGRP-deficient rats when compared with controls. The infusion of IGF-1 resulted in an equivalent decrease in MAP in both the CGRP-deficient and control rats. IGF 1 infusion did not change the heart rate in control rats but decreased it in CGRP deficient rats. IGF-1 also increased flow as determined by conductance in the iliac, renal, and superior mesenteric vascular beds in both vehicle controls and CGRP-deficient rats. We concluded that unlike insulin the IGF-1-mediated vasodilatory response is not mediated by the CGRP-dependent perivascular neurons. PMID- 12588047 TI - Estrogen-induced gonadotropin surge in rhesus monkeys is not inhibited by cortisol synthesis inhibition or hypoglycemia. AB - Acute administration of corticotrophin-releasing hormone (CRH) has been shown to inhibit gonadotropin secretion in several species including rodents, sheep, humans, and nonhuman primates. Similarly, a variety of acute stressors have been shown to inhibit tonic gonadotropin secretion and may do so through a CRH mechanism. Stress-induced inhibition of tonic gonadotropin secretion below levels required for follicular maturation would be expected to inhibit ovulation. An additional mechanism whereby acute stressors could interfere with ovulation is through inhibition of the preovulatory gonadotropin surge. In the present study, we determined the effect of acute activation of the hypothalamic-pituitary adrenal (HPA) axis on phasic gonadotropin secretion in female rhesus monkeys. Activation of the HPA axis was achieved by either a hypoglycemic challenge or blockage of cortisol synthesis with metyrapone, 24 h after an estradiol benzoate challenge. Neither metyrapone nor insulin-induced hypoglycemia inhibited gonadotropin secretion. In fact, the initiation of the luteinizing hormone and follicle-stimulating hormone surge was advanced by 7.4 +/- 0.4 h (p < 0.001) and 4.8 +/- 1.4 h (p = 0.04) respectively, in metyrapone-treated monkeys compared with saline controls. By contrast, hypoglycemia did not affect the gonadotropin surge. The gonadotropin surge was preceded by increased progesterone secretion in metyrapone-treated but not insulin-treated monkeys. This difference in progesterone secretion likely explains the advancement of the gonadotropin surge in the metyrapone-treated animals. PMID- 12588049 TI - Polymorphism in intron 2 of islet amyloid polypeptide gene is associated with lower low-density lipoprotein cholesterol in nondiabetic subjects and in type 2 diabetic patients. AB - The aim of this study was to investigate the presence of mutations in the islet amyloid polypeptide (IAPP) gene in a Spanish population with type 2 diabetes and gestational diabetes mellitus (GDM). Using polymerase chain reaction single stranded conformation polymorphism, we examined the coding region and the 5' untranslated region (UTR) of the IAPP gene in 177 unrelated type 2 diabetic patients, 110 healthy control subjects, 38 women with GDM, and 38 gestational control subjects. Mutations were confirmed by DNA sequencing. A heterozygous C-to A nucleotide substitution at +79 bp in intron 2 of the IAPP gene was detected. The frequencies of the +79-bp polymorphism (A allele) were 6.8% in type 2 diabetic patients, 7.7% in nondiabetic control subjects, 11.8% in women with GDM, and 9.2% in gestational control subjects. No AA genotypes were detected. Nondiabetic subjects and patients with type 2 diabetes bearing the CA genotype had lower low-density lipoprotein (LDL) cholesterol levels than subjects bearing wild genotype. Multivariate logistic regression analysis showed an independent association (p < 0.001; odds ratio: 0.33; 95% confidence interval: 0.17-0.63). We did not detect any sequence variant within exons 1 or 2. One diabetic patient was heterozygous for a silent mutation at codon 31 of exon 3 (Asn31 AAC --> AAT). Our findings indicate that the presence of the +79-bp polymorphism of the IAPP gene in nondiabetic subjects and in patients with type 2 diabetes is associated with lower levels of LDL cholesterol. Furthermore, abnormalities of the coding regions or the 5'-UTR of the IAPP gene are not associated with type 2 diabetes or GDM in the Spanish population. PMID- 12588048 TI - Oral tungstate treatment improves only transiently alteration of glucose metabolism in a new rat model of type 2 diabetes. AB - It has been shown that tungstate is an effective hypoglycemic agent in several animal models of diabetes. In this study, we examined the effectiveness of oral tungstate treatment in a new experimental diabetic syndrome, induced by streptozotocin (STZ) and nicotinamide in adult rats, that shares several features with human type 2 diabetes. Sodium tungstate was administered in the drinking water (2 mg/mL) of control and diabetic rats for 15, 30, 60, and 90 d. Glucose metabolism was explored in vivo by intravenous glucose tolerance test. Insulin secretion and action were assessed in vitro in the isolated perfused pancreas and isolated adipocytes, respectively. Two weeks of tungstate treatment did not modify the moderate hyperglycemia of diabetic rats but reduced their intolerance to glucose, owing to an enhancement of postloading insulin secretion. However, this effect was transient, since it declined after 30 d and vanished after 60 and 90 d of tungstate administration, whereas a trend toward a reduction in basal hyperglycemia was observed on prolonged treatment. Oral tungstate was unable to modify glucose-stimulated insulin secretion in the isolated perfused pancreas, as well as muscle glycogen levels, hepatic glucose metabolism, and insulin stimulated lipogenesis in isolated adipocytes. Nevertheless, the decreased insulin content of pancreatic islets of diabetic rats was partially restored on prolonged tungstate treatment. In conclusion, in the STZ-nicotinamide model of diabetes, tungstate was unable to permanently correct the alterations in glucose metabolism, despite some indirect evidence of a trophic effect on beta-cells. The ineffectiveness of tungstate could be related to the absence, in this diabetic syndrome, of relevant metabolic alterations in the liver, which thus appear to constitute the major target of tungstate action. PMID- 12588050 TI - Decreased blood activity of glucose-6-phosphate dehydrogenase associates with increased risk for diabetes mellitus. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency predisposes affected individuals highly susceptible to oxidative stress, which is one of the risk factors for diabetes. To evaluate the relationship between blood level of G6PD activity and diabetes in Taiwan, blood G6PD activity was analyzed among 237 patients with diabetes and 656 healthy subjects. A significant difference in the distribution of G6PD activities as grouped by an increment of 100 U/10(12) red blood cells (RBCs) was observed between diabetic patients and healthy subjects. The odds ratio for diabetes was 1.46 (95% confidence interval = 1.11-1.92) for every decrement of 100 U/10(12) RBC G6PD activities in these subjects. The data indicate that low G6PD activity is another risk factor for diabetes. PMID- 12588052 TI - Regulation of uncoupling protein-2 mRNA in L6 myotubules: II: Thyroid hormone amplifies stimulation of uncoupling protein-2 gene by thiazolidinediones and other peroxisome proliferator-activated receptor ligands in L6 myotubules: evidence for a priming effect. AB - The stimulation of the uncoupling protein-2 gene (ucp2) by thyroid hormone (triiodothyronine [T3]) in vivo is variable, suggesting complex interactions and even the possibility of indirect effects. We investigated the effect of T3 on ucp2 expression in L6 myotubules. Alone, T3 did not significantly stimulate ucp2 expression in L6 cells, but it amplified the stimulation by thiazolidinediones (TZDs). L6 cells expressed both alpha1 and beta1 thyroid hormone receptors and the data were consistent with the effect being mediated by these receptors. T3 also enhanced the stimulation of ucp2 by the nonselective peroxisome proliferator activated receptor (PPAR) ligands bezafibrate and carbacyclin, but not that by oleic acid or norepinephrine. L6 cells expressed PPARbeta and PPARgamma, but not PPARalpha. As short as a 1-h preexposure of L6 cells to T3 was sufficient to amplify the effect of PPAR ligands. Neither transcription nor translation was needed for this effect of T3. T3 did not affect the t1/2 of UCP2 mRNA. The histone deacetylases inhibitor trichostatin A (TSA) stimulated the expression of ucp2 but did not add to the effect of T3 nor did this hormone enhance the effect of TSA. These results suggest that T3 selectively enhances the transcriptional stimulation of ucp2 by TZDs and nonselective PPAR ligands by priming the gene to a transactivating signal(s) generated by such ligands. PMID- 12588051 TI - Regulation of uncoupling protein-2 mRNA in L6 myotubules: I: Thiazolidinediones stimulate uncoupling protein-2 gene expression by a mechanism requiring ongoing protein synthesis and an active mitogen-activated protein kinase. AB - The mitochondrial uncoupling protein-2 (UCP2) can uncouple phosphorylation to subserve several functions. It has been reported that the insulin sensitizers, thiazolidinediones (TZDs), increase UCP2 mRNA levels and, more recently, that TZDs stimulate UCP2 reporter genes but that the sequences involved do not bind peroxisome proliferator-activated receptor gamma (PPARgamma). We report here that TZDs stimulated UCP2 gene (ucp2) transcription in L6 myotubules involving an indirect mechanism. L6 cells contained comparatively small amounts of PPARgamma mRNA but clearly detectable amounts of PPARgamma2 protein. UCP2 mRNA levels were increased in a time- and concentration-dependent manner by TZDs. UCP2 mRNA had slow turnover (t 1/2 approximately 38 h), and this was not affected by TZDs. Bisphenol A diglycidyl ether, a PPARy antagonist, concentration dependently inhibited the TZD-induced increase in UCP2 mRNA. Blockade of protein synthesis with cycloheximide as well as abrogation of mitogen-activated protein kinase (MAPK) activity with PD98059 or U0126 also prevented the TZD-induced increase in UCP2 mRNA. As with autologous UCP2 mRNA, TZDs stimulated reporter gene expression directed by ucp2 sequences in transiently transfected L6 cells. The effect was enhanced by cotransfection of PPARgamma + retinoid X receptor gamma and prevented by MEK blockade. TZDs, however, did not increase the activation of MAPK, nor did its activation by other means (change of medium, insulin-like growth factor-1, insulin) increase UCP2 mRNA, indicating that phosphorylation is not limiting. These results suggest that TZDs indirectly stimulate ucp2 transcription by inducing-via PPARgamma-limiting amounts of a protein, which must be phosphorylated by MAPK to stimulate the gene. PMID- 12588053 TI - Nutritional interventions for individuals with bulimia nervosa. AB - Many physical and psychological effects of bulimia nervosa are caused by the patient's partial starvation and chaotic nutritional cycle. Attention should thus be initially directed to correcting nutritional deficiencies and abnormal eating patterns, and providing dietary counselling. Nevertheless, very little has been written about the nutritional management of this eating disorder. Nutritional counselling for bulimia patients is reviewed in this paper. Current knowledge about nutritional therapy and its efficacy, goals and objectives is presented, along with recommendations used in treatment programmes. Lastly, the key steps of nutritional management are summarised. PMID- 12588054 TI - Interdisciplinary treatment of a female outpatient population. Organizational model and preliminary results. AB - OBJECTIVE: To compare the efficacy of diet alone vs interdisciplinary therapy. METHOD: 256 women were divided into an interdisciplinary therapy group composed of 102 subjects and a control group composed of 154 subjects treated by means of diet alone. RESULTS: 21.56% of the interdisciplinary group did not come for the 2 check-ups compared with 49.35% of the diet group (p < 0.001). A weight loss of 5.88 kg and 4.18 kg for the interdisciplinary and the diet group was recorded 8 weeks after the start of the programme (second check-up). DISCUSSION: Interdisciplinary therapy is necessary to personalize treatment and satisfy the patient's desire and need for appropriate and effective management. PMID- 12588056 TI - The influence of definitions on the prevalence of eating problems in an adolescent population. AB - OBJECTIVE: The definitions of eating problems vary widely as they integrate cognitive, behavioural and physiological components to different degrees. The aim of this study was to show how much these differences affect the prevalence of eating problems in an adolescent population. METHOD: All of the 9131 adolescents in a Norwegian county participated in a health study, 8042 (88.1%) of whom gave valid responses to a questionnaire covering various aspects of eating problems, and had their weight and height measured. RESULTS: The prevalence of eating problems ranged from 0.3 to 47.0% depending on the definitions used. The various definitions also gave different gender ratios, although the prevalences increased with age in females only. CONCLUSIONS: The various definitions of eating problems had a low degree of correlation and led to quite variable prevalences. All of them showed that the prevalence of eating problems was higher in females, but their prevalence in males was also considerable. PMID- 12588055 TI - Weight and eating concerns in outpatient men and women being treated for substance abuse. AB - This study examined the specific features of weight and eating concerns, and explored gender differences, in a racially diverse group of outpatients (45 men and 39 women) currently being treated for substance abuse but abstinent from substance use. Overweight was prevalent and similarly distributed across gender: 60% of the men and 69% of the women were overweight. Most of the overweight participants did not identify themselves as such. No significant gender differences were observed in terms of the behavioural features of eating disorders: 8% of the men and 11% of the women reported binge eating on at least one day per week; inappropriate weight compensatory behaviours were reported by 8% of the men vs 16% of the women, but the women had significantly higher attitudinal scores than the men. Our findings suggest that, in patients recovering from substance abuse, overweight and features of eating disorders are common in both men and women, and occur in poor and minority groups. PMID- 12588057 TI - Hospitalized anorexics and resistance training: impact on body composition and psychological well-being. A preliminary study. AB - Ritualised exercise commonly accompanies the clinical presentation of anorexia nervosa (AN) sufferers, but there is a paucity of research on structured exercise as a therapeutic intervention. This study examined the usefulness of resistance training as part of a treatment programme for hospitalized anorexics. The study sample consisted of 21 subjects: seven anorexics participating in the exercise programme, seven non-participating anorexics, and seven subjects unaffected by an eating disorder but who participated in the exercise programme. The results show that resistance training is associated with an improvement in body composition and psychological well-being and, although it does not confer an outcome advantage in this regard, it seems to be a useful adjunct in the treatment of hospitalized anorexics. PMID- 12588058 TI - What happens to eating disorder outpatients who withdrew from therapy? AB - OBJECTIVE: Dropouts are frequent among eating disorder (ED) patients, but less is known about their natural history. This paper assesses the outcome of outpatients who dropped out from a therapy programme and its possible causes. MATERIAL AND METHODS: From 1992 to 1994, we assessed 222 ED subjects. Psychiatrists expert in EDs evaluated these subjects by defining baseline parameters and diagnosis was made according to the 3rd revisioned edition of the Diagnostic and Statistical Manual of Mental Disorders. One hundred and twenty-eight subjects (57%) dropped out during the treatment. In 1997, we contacted them, reassessed the same baseline parameters and asked for a self-judgment about their social and clinical condition during the previous 2-5 years. Patients were classified as "improved" and "not improved" (stationary or worse) according to their social, physical and psychological condition. The relation between baseline condition and outcome was determined statistically. RESULTS: Seventy-one percent of subjects were "improved" and no deaths were recorded. A significant correlation was found between "duration of illness" and no treatment following a dropout. DISCUSSION: The high percentage of improvement among dropouts was unexpected. Shorter duration of illness and lack of specific therapy in the improved patients suggest the existence of a subset of ED patients with acute onset and a spontaneous tendency to improve. This point obviously requires further investigation. PMID- 12588059 TI - A survey of anorexia nervosa using the Arabic version of the EAT-26 and "gold standard" interviews among Omani adolescents. AB - OBJECTIVE: The purpose of this study was to assess the validity of the Eating Attitude Test (EAT) in identifying the presence and severity of eating pathology in male and female Omani urban adolescents and to establish cut-off scores that matched those of anorexia identified by gold standard interviews without fear of fatness criteria. METHODS: Both females (n=126) and males (n=136) were screened using the Arabic version of the EAT-26 and interviewed using a semi-structured, Composite International Diagnostic Interview (CIDI) in order to investigate the relationship between false positives and false negatives at various EAT-26 cut off points. A receiver operating characteristics (ROC) curve was calculated to discriminate the power of the EAT-26 for every possible threshold score. RESULTS: The EAT-26 identified 29% of the subjects as probable anorexic cases as against 9.5% identified during the structured interview based on the anorexia gold standard (32% males and 68% females). The sensitivity and specificity of the EAT 26 were respectively 24% and 69.6%. When using the ROC curve, a cut-off score of 10 gave the best compromise between sensitivity (64%) and specificity (38%). DISCUSSION: Although the EAT-26 is the most widely used screening instrument in cross-cultural studies, it does not appear to be reliable in identifying probable cases of anorexia among Omani adolescents. The use of a gold standard interview without fat phobia criteria indicated that the rate of anorexia nervosa may be more prevalent among males than previously estimated. This intriguingly high preponderance of males is discussed in terms of prevailing demographic trends in Oman. PMID- 12588060 TI - An examination of perceptual and cognitive set shifting tasks in acute anorexia nervosa and following recovery. AB - OBJECTIVES: People with anorexia nervosa have a broad spread of symptoms including disturbances in perception, cognition, emotions and behaviour. The aim of this study was to examine tests of executive function (in particular, perceptual and cognitive set shifting tasks) in patients with a current or past diagnosis of anorexia nervosa (AN). METHOD: 30 AN patients, 16 subjects recovered from anorexia nervosa (ANR) and 23 healthy controls were examined using tests of executive function (initiation tasks and perceptual and cognitive set shifting). RESULTS: The AN and ANR subjects had significantly higher perceptual and cognitive set shifting scores than the controls. CONCLUSIONS: Impaired executive function in terms of set shifting tasks could represent a vulnerability factor for AN. PMID- 12588061 TI - Body size dissatisfaction among male and female triathletes. AB - Body size dissatisfaction was assessed among 583 male and female triathletes residing in the USA. The males perceived themselves to be larger than their calculated body mass index (BMI discrepancy index: -1.92) and desired to be larger (BMI discrepancy index: -0.7), whereas the females perceived themselves to be smaller than their calculated BMI (BMI discrepancy index: 1.67) and desired to be much smaller (BMI discrepancy index: 3.61). The male triathletes categorised as underweight showed no discrepancy between their perceived and desired BMI, whereas 10% of the females in the underweight category desired to be larger than perceived and 13% desired to be smaller. Among the males whose BMI fell into the healthy range, 8.0% indicated a desire to be larger than perceived and 19.3% desired to be smaller; the majority of female triathletes in the healthy BMI category desired to be smaller, with only 2.4% desiring to be larger. PMID- 12588062 TI - The cumulative impact of being overweight on women's body esteem: a preliminary study. AB - As much research indicates that a woman's current weight status is an important predictor of her body esteem, we examined the relationship between weight status history (retrospective self-report since the age of 5 years) and current body esteem among 266 female college students, who completed a measure of body esteem and reported whether they had been "underweight", "normal weight", or "overweight" during 3 age ranges. Analysis of variance (ANOVA) was used to examine differences in current body esteem between groups characterised by the following weight history trajectories: "always overweight", "always normal/underweight", "increase in weight over time" and "decrease in weight over time". Although the women in the groups "always overweight" and "increase in weight over time" were currently overweight [according to self-reported body mass index (BMI)], the current body esteem of the former was significantly lower (worse) than that of the latter. We introduce the idea that being overweight for a long period of time has a cumulatve negative impact on adult body esteem. PMID- 12588063 TI - Attachment theory, metacognitive functions and the therapeutic relationship in eating disorders. AB - J. Bowlby's attachment theory is used to explore a fundamental motivational force in human behaviour, namely the search for physical and emotional protective intimacy according to an integrated viewpoint remote from Freud's drive-based approach. This theory has received contributions from ethology, the neurosciences, psychoanalysis, cognitive psychology and evolutionary epistemology. Attachment patterns or styles express both the mental and behavioural strategies adopted by individuals to cope with their attachment needs, and their relational history in this area. Experiences of adequate quality promote metacognitive functions, namely a set of mental processes whose role is to protect the individual from psychopathological suffering and construct and articulate a sense of self. Attachment theory and metacognition are the underlying assumptions of reflections on the dynamics marking the development of sometimes very difficult relationships between therapists and eating disorder (ED) patients, who are being increasingly classed as "severe patients" on account of their interpersonal characteristics. PMID- 12588064 TI - Disability pension for a middle-aged woman with anorexia nervosa. PMID- 12588065 TI - Calcaneal ultrasonography for bone assessment. PMID- 12588066 TI - Vitamin D as a risk reduction factor for colorectal cancer. PMID- 12588067 TI - Managing asthma should not focus on dust mite control. PMID- 12588068 TI - Guidelines for proper use of child safety seats. PMID- 12588069 TI - Dermatitis herpetiformis and a gluten-free diet. PMID- 12588070 TI - What is the most effective approach to chronic pain? PMID- 12588071 TI - Medical needs of foster children. PMID- 12588072 TI - Are low-fat diets better than other weight-reducing diets in achieving long-term weight loss? PMID- 12588073 TI - Common dental emergencies. AB - Dental caries, a bacterial disease of teeth characterized by destruction of enamel and dentine, is often the underlying cause of dental pain. When a carious lesion impinges on the dental pulp, pulpitis follows and, ultimately, necrosis of the pulp occurs. Untreated necrosis may lead to a localized abscess or a spreading infection into the surrounding soft tissue that results in cellulitis. Immediate treatment involves antibiotic therapy for cellulitis, perhaps with drainage of abscesses, while definitive treatment requires root canal therapy or extraction of the involved tooth. Pericoronitis is an inflammation of the soft tissue overlying a partially erupted tooth. Localized cases respond to irrigation. Secondary cellulitis can develop. Definitive treatment may require surgical extraction of the underlying tooth or excision of the gum flap. Avulsion of a permanent tooth secondary to trauma is a true dental emergency. The tooth should be reimplanted on the spot, and the patient should be seen immediately by a dentist for splinting and antibiotic prophylaxis. Most dental problems can be prevented with regular dental care and steps to minimize risks of oral trauma. PMID- 12588074 TI - Cytomegalovirus. AB - Cytomegalovirus (CMV) is a prevalent viral pathogen. The majority of persons with acute CMV will experience an inapparent infection. Primary CMV infection will cause up to 7 percent of cases of mononucleosis syndrome and will manifest symptoms almost indistinguishable from those of Epstein-Barr virus-induced mononucleosis. CMV, or heterophil-negative mononucleosis, is best diagnosed using a positive IgM serology. Complications of acute CMV infection in immunocompetent persons are rare, except in newborns. The virus usually is spread through close personal contact; transmission risk can be reduced by following simple hygienic and handwashing techniques. Severe illness can occur after reactivation of the latent virus in immunosuppressed persons. The retina is the most common site of CMV-induced pathology in persons with human immunodeficiency virus infection. Advances in the treatment of human immunodeficiency virus infection with highly active antiretroviral therapy (HAART) have decreased the incidence of CMV retinitis but have resulted in a new set of ophthalmologic complications induced by restoration of immune competency and the pro-inflammatory response of the patient to CMV. If HAART restores the patient's CD4 cell count to above 100 to 150 per mm3 (100 to 150 x 10(6) per L), it may preclude lifelong treatment for CMV retinitis. PMID- 12588075 TI - Information from your family doctor. Cytomegalovirus. PMID- 12588076 TI - Hip fractures in adults. AB - Patients with hip fracture typically present to the emergency department or their physician's office after a fall. They are often unable to walk, and they may exhibit shortening and external rotation of the affected limb. Frequently, they have hip pain. In some instances, however, patients with hip fracture may complain only of vague pain in their buttocks, knees, thighs, groin, or back. Their ability to walk may be unaffected, and initial radiographic findings may be indeterminate. In these patients, additional studies, such as magnetic resonance imaging or bone scanning, may be necessary to confirm the presence of hip fracture. A high index of suspicion often is required for prompt diagnosis and treatment of an occult hip fracture. Even when a patient is able to walk and has no documented trauma, localized hip pain, or typical shortening and malrotation deformity, the family physician should be alert to the possibility of hip fracture, particularly in a patient who is older than 65 years, presents with nonspecific leg discomfort, and complains of difficulty bearing weight on the affected limb. A heightened suspicion for hip fracture should lead to further diagnostic evaluation, especially if the patient has additional risk factors, such as use of a complicated drug regimen, impaired vision, physical or neurologic impairment, or comorbid condition (e.g., osteoporosis, malignancy). When hip fracture is detected early, appropriate treatment can minimize morbidity and mortality and prevent the rapid decline in quality of life that often is associated with this injury. PMID- 12588077 TI - Antidepressants: update on new agents and indications. AB - A number of antidepressants have emerged in the U.S. market in the past two decades. Selective serotonin reuptake inhibitors have become the drugs of choice in the treatment of depression, and they are also effective in the treatment of obsessive-compulsive disorder, panic disorder, and social phobia. New indications for selective serotonin reuptake inhibitors include post-traumatic stress disorder, premenstrual dysphoric disorder, and generalized anxiety disorder. Extended-release venlafaxine has recently been approved by the U.S. Food and Drug Administration for the treatment of generalized anxiety disorder. Mirtazapine, which is unrelated to the selective serotonin reuptake inhibitors, is unique in its action--stimulating the release of norepinephrine and serotonin. The choice of antidepressant drug depends on the agent's pharmacologic profile, secondary actions, and tolerability. Sexual dysfunction related to the use of antidepressants may be addressed by reducing the dosage, switching to another agent, or adding another drug to overcome the sexual side effects. Augmentation with lithium or triiodothyronine may be useful in patients who are partially or totally resistant to antidepressant treatment. Finally, tapering antidepressant medication may help to avoid discontinuation syndrome or antidepressant withdrawal. PMID- 12588079 TI - Information from your family doctor. Thyroid nodules. PMID- 12588078 TI - Thyroid nodules. AB - Palpable thyroid nodules occur in 4 to 7 percent of the population, but nodules found incidentally on ultrasonography suggest a prevalence of 19 to 67 percent. The majority of thyroid nodules are asymptomatic. Because about 5 percent of all palpable nodules are found to be malignant, the main objective of evaluating thyroid nodules is to exclude malignancy. Laboratory evaluation, including a thyroid-stimulating hormone test, can help differentiate a thyrotoxic nodule from an euthyroid nodule. In euthyroid patients with a nodule, fine-needle aspiration should be performed, and radionuclide scanning should be reserved for patients with indeterminate cytology or thyrotoxicosis. Insufficient specimens from fine needle aspiration decrease when ultrasound guidance is used. Surgery is the primary treatment for malignant lesions, and the extent of surgery depends on the extent and type of disease. Ablation by postoperative radioactive iodine is done for high-risk patients--identified as those with metastatic or residual disease. While suppressive therapy with thyroxine is frequently used postoperatively for malignant lesions, its use for management of benign solitary thyroid nodules remains controversial. PMID- 12588080 TI - Depression in children and adolescents. PMID- 12588081 TI - Photo quiz. Woody edema of the legs. PMID- 12588082 TI - AHA releases scientific statement on cardiovascular health in childhood. American Heart Association. PMID- 12588083 TI - A mother who refuses to vaccinate her child. PMID- 12588084 TI - Postmenopausal breast cancer is associated with high intakes of omega6 fatty acids (Sweden). AB - OBJECTIVE: To estimate the postmenopausal breast cancer risk associated with total fat intake, different types and relative proportions of dietary fat using a nested, matched case-control study within the Malmo Diet and Cancer (MDC) cohort, Sweden. METHODS: Among women 50 years or older at baseline (n = 12,803), incident breast cancer cases (n = 237) were matched to controls (n = 673) on age and screening date. Data were obtained by a "novel" diet history method, a structured questionnaire, and anthropometric measurements. Conditional logistic regression examined breast cancer risks associated with quintiles of fat intake residuals adjusted for energy and potential confounders. RESULTS: Saturated fat and the omega3-omega6 fatty acid ratio were not related to increased risks, but positive trends were seen for total (p = 0.031), monounsaturated (p = 0.002), and polyunsaturated fat (p = 0.0009), especially omega6 fatty acids and the polyunsaturated-saturated fat ratio (p = 0.004). With mutual adjustment for different types of fat, an elevated risk remained significant in the highest omega6 fatty acid quintile (RR= 2.08, 95% CI 1.08-4.01). CONCLUSIONS: Postmenopausal breast cancer was positively associated with total, monounsaturated, and polyunsaturated fat. However, with mutual adjustment for other types of fat, specifically high intakes of omega6 fatty acids were associated with an increased risk. PMID- 12588085 TI - Testicular cancer and electromagnetic fields (EMF) in the workplace: results of a population-based case-control study in Germany. AB - OBJECTIVES: In a population-based case-control study, we examined the association of testicular cancer and electromagnetic fields (EMF) in the workplace. METHODS: Incident cases (n = 269) were recruited between 1995 and 1997. A total of 797 controls matched on age and region were randomly selected from mandatory registries of residents. EMF exposure was assessed for five categories in standardized face-to-face interviews using closed questions. For each exposure category, odds ratios (OR) were calculated, stratified by age and region, and in a more complex model weighted by duration and distance using conditional logistic regression. Subgroup analyses were conducted for seminoma and non-seminoma and for blue- and white-collar workers. Additionally, potential radar exposure was individually assessed by experts based on all available information including free text. RESULTS: There was no excess risk for cases who reported to have ever worked near the following: radar units (OR = 1.0; 95% CI = 0.60-1.75); radiofrequency emitters (OR = 0.9; 95% CI = 0.60-1.24); electrical machines (OR = 1.0; 95% CI = 0.72-1.33): high-voltage lines or high-voltage electrical transmission installations (OR = 0.7; 95% CI = 0.38-1.18); or visual display units or complex electrical environments (OR = 0.9; 95% CI = 0.67-1.21). The results for the weighted exposure and subgroup analyses did not differ substantially. For radar exposure as assessed by the experts, the OR was 0.4 (95% CI = 0.13-1.16). CONCLUSIONS: EMF exposure in the workplace does not seem to be a relevant risk factor for testicular cancer in our study. PMID- 12588086 TI - Using hierarchical spatial models for cancer control planning in Minnesota (United States). AB - OBJECTIVE: Region-specific maps of cancer incidence, mortality, late detection rates, and screening rates can be very helpful in the planning, targeting, and coordination of cancer control activities. Unfortunately, past efforts in this area have been few, and have not used appropriate statistical models that account for the correlation of rates across both neighboring regions and different cancer types. In this article we develop such models, and apply them to the problem of cancer control in the counties of Minnesota during the period 1993-1997. METHODS: We use hierarchical Bayesian spatial statistical methods, implemented using modern Markov chain Monte Carlo computing techniques and software. RESULTS: Our approach results in spatially smoothed maps emphasizing either cancer prevention or cancer outcome for breast, colorectal, and lung cancer, as well as an overall map which combines results from these three individual cancers. CONCLUSIONS: Our methods enable us to produce a more statistically accurate picture of the geographic distribution of important cancer prevention and outcome variables in Minnesota, and appear useful for making decisions regarding targeting cancer control resources within the state. PMID- 12588087 TI - Relation of impaired glucose tolerance and diabetes mellitus to colorectal adenomas in Japan. AB - OBJECTIVE: Because a link between hyperinsulinemia and colorectal carcinogenesis has been hypothesized, we explored the relation between glucose tolerance status and colorectal adenomas. METHODS: We conducted a case-control study of 560 cases of histologically confirmed colorectal adenomas and 829 controls of normal total colonoscopy in Japanese middle-aged men. Subjects were classified into normal, impaired fasting glucose (IFG), impaired glucose tolerance (IGT), newly diagnosed type 2 diabetes mellitus (DM), and known DM based on fasting blood glucose, 75 g glucose tolerance test, and medical history. Odds ratios (ORs) and 95% confidence intervals (CIs) were obtained from logistic regression analysis with subjects with normal glucose tolerance and IFG as a referent group. RESULTS: IGT, new type 2 DM, and known DM were each associated with a modest increase in the risk of colorectal adenomas after adjustment for possible confounding factors. An increased risk of large, but not small, adenomas was observed among men with IGT and new type 2 DM. An increased risk associated with new type 2 DM was observed for proximal adenomas only. CONCLUSIONS: Not only type 2 DM but also IGT is associated with increased risk of colorectal adenomas. Hyperinsulinemia may be involved in colorectal carcinogenesis. PMID- 12588088 TI - Toenail selenium concentration and lung cancer in male smokers (Finland). AB - BACKGROUND: The objective of this investigation was to evaluate the association between toenail selenium concentration and lung cancer risk in male smokers. METHODS: We conducted a nested case-control study within the Alpha-Tocopherol Beta-Carotene Cancer Prevention Study cohort. This substudy included 250 randomly selected incident lung cancer cases and 250 controls matched on age (up to +/- 5 years), intervention group assignment, and date of randomization (+/- 15 days). Odds ratios (ORs) and 95% confidence intervals (CIs) were determined using conditional logistic regression methods. Finland began fortification of agricultural fertilizers in the fall of 1984, increasing the dietary intake, plasma, and toenail selenium concentrations for the population. The present analyses were based on the calculated residual of toenail selenium after regressing it on date of randomization. The selenium residual and the interaction of the residual with date of randomization were included in models with smoking status and body mass index as covariates. RESULTS: We observed a suggestion of a protective association for higher selenium status among men who entered the trial early (when the range of selenium values included very low levels). The OR for men with adjusted toenail selenium concentrations at the 75th percentile compared to those with the lowest selenium concentrations ranged between 0.20 (0.09-0.44) for men randomized earliest in the trial and 0.61 (0.27-1.41) for men randomized in the fifth year. CONCLUSIONS: These results suggest that low selenium status may be associated with increased risk for lung cancer. PMID- 12588089 TI - Effect of diet and exercise on serum insulin, IGF-I, and IGFBP-1 levels and growth of LNCaP cells in vitro (United States). AB - OBJECTIVE: Accumulating evidence indicates that prostate cancer is associated with high levels of serum IGF-I. This study was conducted to determine whether a low-fat diet and exercise (DE) intervention may modulate the IGF axis and reduce prostate cancer cell growth in vitro. METHODS: Fasting serum was obtained from 14 men (age 60 +/- 3 years) participating in an 11-day DE program and from eight similarly aged men who had followed the DE program for 14.2 +/- 1.7 years (long term). Insulin, IGF-I, IGFBP-1, and IGFBP-3 were measured by ELISA, and serum was used to stimulate LNCaP cell growth in vitro. RESULTS: Serum IGF-I levels decreased by 20% while IGFBP-1 increased by 53% after 11-day DE. In the long-term group, IGF-I was 55% lower, while IGFBP-1 was 150% higher relative to baseline. Serum insulin decreased by 25% after 11-day DE and was 68% lower in the long-term group, relative to baseline. No changes in serum IGFBP-3 were observed. Serum stimulated LNCaP cell growth was reduced by 30% in post-11-day serum and by 44% in long-term serum relative to baseline. LNCaP cells incubated with post-DE serum showed increased apoptosis/ necrosis, compared to baseline. CONCLUSIONS: A low fat diet and exercise intervention induces in-vivo changes in the circulating IGF axis and is associated with reduced growth and enhanced apoptosis/necrosis of LNCaP tumor cells in vitro. PMID- 12588090 TI - Increased risk of early-stage breast cancer related to consumption of sweet foods among women less than age 45 in the United States. AB - OBJECTIVES: To evaluate the associations of dietary macronutrients, food groups, and eating patterns with risk of breast cancer in a population-based case-control study. METHODS: In this study among women 20-44 years of age, 568 cases with breast cancer and 1451 population-based controls were included. They completed a detailed in-person interview, a self-administered food-frequency questionnaire and were measured for anthropometric indices. Logistic regression was used to estimate odds ratios (OR) and their 95% confidence intervals (CI) of breast cancer, adjusted for age, study site, race, education, alcohol consumption, oral contraceptive usage, smoking status, and body mass index. RESULTS: There was no association between breast cancer risk and intake of calories, macronutrients, or types of fat. Risk of breast cancer was unrelated to intakes of a variety of food groups, including red meats, dairy, high-fat snacks and desserts, or foods high in animal fat. Increased risk was observed for high intake of a food group composed of sweet items, particularly sodas and desserts. Risk increased linearly with percent of calories from sweets and frequency of sweets intake. Consumption of sweets 9.8 or more times per week compared with <2.8 times per week was associated with an adjusted OR of 1.32 (95% CI = 1.0-1.8). This association did not appear to be due to the high-fat foods or carbonated beverages that comprised the food group. Compared with women reporting one or two meals and snacks per day, reduced risks were noted for women reporting six or more (OR = 0.69, 95% CI = 0.4-1.1). CONCLUSIONS: These data suggest a modest relationship between intakes of sweet items with risk of in-situ and localized breast cancer in young women. This relation is consistent with the hypothesized link of high insulin exposure and risk of breast cancer. There was some suggestion that women who ate many times during the day were at reduced risk of disease, which is also consistent with an insulin-related mechanism. PMID- 12588091 TI - Urinary estrogen metabolites and prostate cancer: a case-control study in the United States. AB - OBJECTIVE: To analyze the association of prostate cancer risk with estrogen metabolism, expressed as the ratio of 2-hydroxyestrone (2-OHE1) to 16alpha hydroxyestrone (16alpha-OHE1), in a case-control study conducted in Buffalo, NY, between 1998 and 2001. METHODS: One hundred and thirteen men, aged 45-85 years, with incident, primary pathologically confirmed prostate cancer were enrolled in the study; 317 residence-matched controls were also enrolled. Cases were enrolled and the specimens collected before starting any therapy. To exclude latent prostate carcinomas, the present study included only patients with clinically apparent disease (stage B and higher). Prostate cancer cases and control subjects were excluded if on hormonal treatment, or affected with metabolic or endocrine diseases. Control subjects with a prostate-specific antigen (PSA) value higher than 4 ng/ml were excluded from the control group. Urine was used for the determinations of 2-OHE1 and 16alpha-OHE1. Age, race, body weight, waist-to-hip ratio, and smoking were analyzed as possible confounders. RESULTS: Althpugh the results were not statistically significant, elevated 2-OHE1 urinary levels suggested a reduced prostate cancer risk: men in the highest tertile had an adjusted odds ratio (OR) for prostate cancer of 0.83 (95% confidence interval (CI) 0.43-1.44). Conversely, elevated 16alpha-OHE1 urinary levels were associated with an increased risk of prostate cancer: the highest tertile had an adjusted OR for prostate cancer of 1.69 (95% CI 0.93-3.06, p for linear trend = 0.02). Finally, the 2-OHE1 to 16alpha-OHE1 ratio was associated with a reduced risk of prostate cancer with an OR of 0.61 (95% CI 0.33-1.15, p for linear trend = 0.04). CONCLUSION: Results of this case-control study suggest that the estrogen metabolic pathway favoring 2-hydroxylation over 16alpha-hydroxylation may reduce risk of clinically evident prostate cancer. PMID- 12588092 TI - Combined effect of tobacco and alcohol on laryngeal cancer risk: a case-control study. AB - OBJECTIVE: To provide information on the effects of alcohol and tobacco on laryngeal cancer and its subsites. METHODS: This was a case-control study conducted between 1992 and 2000 in northern Italy and Switzerland. A total of 527 cases of incident squamous-cell carcinoma of the larynx and 1297 hospital controls frequency-matched with cases on age, sex, and area of residence were included. Odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were estimated using multiple logistic regression. RESULTS: In comparison with never smokers, ORs were 19.8 for current smokers and 7.0 for ex-smokers. The risk increased in relation to the number of cigarettes (OR = 42.9 for > or = 25 cigarettes/day) and for duration of smoking (OR = 37.2 for > or = 40 years). For alcohol, the risk increased in relation to number of drinks (OR = 5.9 for > or = 56 drinks per week). Combined alcohol and tobacco consumption showed a multiplicative (OR = 177) rather than an additive risk. For current smokers and current drinkers the risk was higher for supraglottis (ORs 54.9 and 2.6, respectively) than for glottis (ORs 7.4 and 1.8) and others subsites (ORs 10.9 and 1.9). CONCLUSIONS: Our study shows that both cigarette smoking and alcohol drinking are independent risk factors for laryngeal cancer. Heavy consumption of alcohol and cigarettes determined a multiplicative risk increase, possibly suggesting biological synergy. PMID- 12588093 TI - Use of anti-inflammatory and non-narcotic analgesic drugs and risk of non Hodgkin's lymphoma (NHL) (United States). AB - OBJECTIVE: To examine whether exposures to anti-inflammatory and non-narcotic analgesic drugs are associated with risk of non-Hodgkin's lymphoma (NHL). METHODS: A case-control study was conducted among women living in upstate New York. The study involved 376 cases of NHL identified through the New York State Cancer Registry and 463 controls randomly selected from the Medicare beneficiary files and New York State driver's license records. Information regarding use of common medications in the past 20 years and potential confounding variables was obtained by telephone interview. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using an unconditional logistic regression model. RESULTS: There were non-significant increases in risk associated with ever use of cortisone injections and oral cortisone (OR = 1.44 (CI 0.98-2.11) for injections and 1.21 (CI 0.73-2.00) for oral cortisone, although there was no clear dose response relationship with either type. On the other hand, the risk of NHL progressively increased with the frequency of use of non-steroidal anti inflammatory and non-narcotic analgesic drugs (NSAID/NNAD) (p-value for trend 0.008). Women who used any of these medications daily for more than 10 years had an OR of 1.90 (CI 1.01-3.57), compared with those who used it less than once a month on average. The risk associated with long-term use was most pronounced for ibuprofen, intermediate for aspirin, and least for acetaminophen. CONCLUSIONS: Because the population-attributable risk associated with NSAID/NNAD use is potentially large, our results need to be verified in further epidemiologic studies. PMID- 12588094 TI - Re: Diagnostic radiation and the risk of multiple myeloma (United States). PMID- 12588095 TI - Green fluorescent protein as a molecular marker in microbiology. AB - Molecular markers such as: lacZ (b-galactosidase), xylE (catechol 2,3 dioxygenase), lux (bacterial luciferase), luc (insect luciferase), phoA (alkaline phosphatase), gusA and gurA (beta-glucuronidase), gfp (green fluorescent protein), bla (beta-lactamase) and other antibiotic resistance markers, heavy metals resistance genes are commonly used in environmental microorganisms research (Errampaii et al., 1998; Kohler et al., 1999). Most of these markers require one or more substrates, complex media and/or expensive equipment for detection. The gfp gene is widely used as a marker because of its very useful properties such as high stability, minimal toxicity, non-invasive detection and the ability to generate the green light without addition of external cofactors and without application of expensive equipment. Various applications of that reporter gene were showed starting from monitoring of microorganism's survival in complex biological systems such as activated sludge to biodegradation of chemical compounds in soil. GFP allowed the detection, determination of spatial location and enumeration of bacterial cells from diverse environmental samples such as biofilm and water. The gfp as a biomarker was very useful in monitoring of gene expression and protein localisation in bacterial cells, too. The techniques with using gfp marker promise to supply a better understanding of environmental processes. It can make possible to use that knowledge in designing more effective and more efficient methods of biodegradation of toxic compounds from different environments. PMID- 12588096 TI - Effect of null mutations in dnaK and dnaJ genes on conjugational DNA transfer, proteolysis and novobiocin susceptibility of Escherichia coli. AB - Escherichia coli null dnaJ and dnaKdnaJ mutants, when introduced to Hfr donor, impair its ability to DNA transfer during conjugation. The additive effect of both mutations was shown. Lack of DnaK and DnaJ chaperones also decrease the extent of proteolysis in mutant strains. This effect is seen only at 42 degrees C. The influence of double dnaKdnaJ deletion but not single dnaJ deletion on novobiocin susceptibility was also demonstrated. PMID- 12588097 TI - Numerical modeling of ferrous-ion oxidation rate in Acidithiobacillus ferrooxidans ATCC 23270: optimization of culture conditions through statistically designed experiments. AB - Statistically designed experimental strategy has been performed in order to evaluate and optimize nutritional and environmental parameters that affect ferrous ion oxidation rate in Acidithiobacillus ferrooxidans ATCC 23270. Plackett Burman design was carried out to evaluate efficiently the biological significance of 10 culture conditions influencing ferrous-ion oxidation rate of A. ferrooxidans grown for 5 days in shake-flask batch mode on the newly modified 9-K media. Among ten fermentation factors examined, the most significant variables influencing ferrous-ion oxidation rate were statistically elucidated to be pH and calcium nitrate as positive contributors, whereas trace metals solution and potassium chloride were the most significant negative contributors. The optimal levels of the most significant three nutritional factors were further predicted from a polynomial model created from the data obtained from three level factorial design, a Box-Behnken design. Predicted optimal ferrous-ion oxidation rate Q(Fe2+) was recorded to be 0.148 (g Fe2+/l/hr). On verifying the predicted value, an experiment was performed under optimal predicted conditions and showed an actual experimental Q(Fe2+) of 0.152 g/l/hr, which was 2.7% over the predicted value. Our optimized medium formula gave overall five folds increase in ferrous ion oxidation rates over the previously published data of standard 9-K medium on batch culture of A. ferrooxidans ATCC 23270 with higher mu(max) (hr(-1)) of 0.177 which was achieved within 75 h incubation in shake-flask culture. PMID- 12588098 TI - Purification and general properties of pectin methyl esterase from Curvularia inaequalis NRRL 13884 in solid state culture using orange peels as an inducer. AB - Pectin methyl esterase (PME) [E.C.3. 1.1.11] production by Curvularia inaequalis (Shear) Boedijn NRRL 13884 was investigated using solid-state culture. The highest level of extracellular pectin methyl esterase was detected with orange peels as an inducing substrate and as a sole carbon source. The enzyme was partially purified using Sephadex G-100 and DEAE-Cellulose column chromatography. It was purified about 40 fold with optimum activity at pH 4.4 and 45 degrees C. The enzyme was activated by Co++, Mg++, Na+, whereas it was slightly activated in the presence of Cu++, K+, Mn++, Zn++. On the other hand Ag++, Ca++ and Hg++ inhibited the activity of the enzyme. The Km was calculated to be 0.52 mM. PMID- 12588099 TI - Ability of Pseudomonas sp. to synthesize aminopeptidases in the presence of various carbon and nitrogen sources. AB - A soil strain of Pseudomonas sp. is able to synthesize at least two aminopeptidases exhibiting high activity in the presence of Phe-beta-NA and Ala beta-NA as substrates. Irrespective of the used substrate, total activity of studied enzymes was strongly related to concentrations of organic components (peptone, glutamic acid, glucose) in mineral media and was the higher, the higher the concentration. Tendency of changes in total activity was similar for alanyl- and phenylalanylaminopeptidase though their response to different concentrations of organic components was different. Specific activity measured in the presence of Phe-beta-NA and Ala-beta-NA as the substrates was not strictly dependent on increasing concentrations of organic components in the media. The highest specific activity of aminopeptidase was obtained in the presence of Phe-beta-NA as a substrate on the fifth day of culture in medium containing 1% glucose. The obtained results seem to indicate the inductive character of the studied aminopeptidases. On the other hand, however, they do not exclude other regulatory mechanisms of their synthesis, including catabolic repression. PMID- 12588100 TI - Primary resistance of Helicobacter pylori to antimicrobial agents in Polish children. AB - Helicobacter pylori resistance to antimicrobial agents is an important factor compromising the efficacy of treatment. Therefore the aims of our study were: to determine the prevalence of H. pylori resistance to clarithromycin, metronidazole, amoxycillin and tetracycline in children prior to eradication therapy, to compare different methods of susceptibility testing and to detect mutations responsible for clarithromycin resistance. During 1996-2000, 259 H. pylori strains were isolated from antral gastric biopsies. Susceptibility to antimicrobials was determined by the agar dilution method and the Etest. Mutations in the 23S rRNA gene associated with clarithromycin resistance were analysed by PCR-RFLP and direct sequencing. Overall, ninety-six strains (37%) were resistant to metronidazole, 50 strains (19.3%) were resistant to clarithromycin, and 20 strains (7.7%) were simultaneously resistant to both drugs. All cultured isolates were sensitive to amoxycillin and only one isolate (0.4%) was resistant to tetracycline. The agar dilution method and the Etest showed a perfect category correlation for clarithromycin and 4% discrepancies for metronidazole. Primary resistance to clarithromycin was mainly associated with an A2143G mutation in the 23S rRNA gene of H. pylori. The study highlights the high prevalence of H. pylori primary resistance to clarithromycin in Polish children, which implies a need for pretreatment susceptibility testing. PMID- 12588101 TI - Antimicrobial activity of substituted 2-trifluoromethyl- and 2 pentafluoroethylbenzimidazoles. AB - Antibacterial and antifungal activity of 2-trifluoromethyl- and 2 pentafluoroethylbenzimidazoles, including a number of newly obtained derivatives, were examined by diffusion method (inhibition area diameter in solid agar medium) and minimum inhibitory concentration (MIC, in liquid and agar medium). Some of the derivatives tested affected fungal colony morphology and exerted genotoxic effects in bacteria. Of the tested compounds, 5,6-dichlorosubstituted derivatives appeared the most active against the majority of microorganisms used. PMID- 12588102 TI - Antifungal activity of Bacillus coagulans against Fusarium sp. AB - The antifungal activity of Bacillus coagulans against three pathogenic species of Fusarium was examined. Fungal growth was determined by colony forming units, dry matter and ergosterol level. Biosynthesis of Fusarium mycotoxins was also investigated. The strongest inhibition of fungal growth was noticed when Bacillus coagulans was co-inoculated at the beginning of culture. Estimation of ergosterol level as a determinant of fungal growth showed the greatest degree of Fusarium sp. inhibition. Addition of Bacillus coagulans to Fusarium culmorum culture inhibits the DON (deoxynivalenol) production. PMID- 12588103 TI - Studies on the effect of inoculation of activated sludge with bacteria actively degrading hydrocarbons on the biodegradation of petroleum products. AB - Eighteen strains of bacteria were isolated from activated sludge purifying petroleum-refining wastewaters. These strains were plated on solidified mineral medium supplemented with oil fraction in concentration 1000 mg/l. Four of the strains that grew best in the presence of oil were selected for further studies. The strains were identified based on Bonde's scheme and microscopic observations. Three of them belonged to the genus Arthrobacter and one to the genus Micrococcus. Stationary cultures of single strains and their mixtures were set up in mineral medium containing oil (sterile and non-sterile) as sole carbon source in concentration 1000 mg/l. The oils were found to be removed the most efficiently by a mixture of the strains. After 14 days of culture the amount of oil was utilized by from 63 to 95%. In the next stage of the studies the bacteria were used to inoculate activated sludge. Stationary cultures of the activated sludge were set up in mineral medium with oil. The utilisation of petroleum products by non-inoculated activated sludge (control), activated sludge inoculated with a single strain or a mixture of all four strains was examined. In both inoculated activated sludge cultures approximately 80% of the oils were removed, compared to 60% in the control activated sludge. Therefore, inoculated activated sludge showed 20% higher effectiveness of removal of petroleum derivatives. PMID- 12588104 TI - Impact of industrial waste water effluents on mycoflora of the shore sediments of the 3rd oxidation pond, with reference to biosorption of heavy metals. AB - The third oxidation pond at 10th of Ramadan desert receives a number of industrial waste water effluents contaminated with the heavy metal ions Zn, Cd, Cu and Ni. The species diversity and fungal community structure of seven different sites at the onshore sediments and offshore were studied. Mycological analysis resulted in isolation of 3912 fungal colonies, 11.7% of this count were recovered from the onshore sediment sites (4 sites) whereas 88.3% were from the offshore sites (3 sites), in the desert. Fungal counts and species diversity at the onshore sites tend to increase with increasing distance far from the waste water input. A complete accordance was observed among the total fungal counts and species variabilities with organic matter content at each sampling site. This relationship was reversed in case of heavy metal contents with both counts and diversity. Seventeen fungal species belonging to seven genera were isolated from all sites under study. Aspergillus spp. constituted the majority of the isolates (51.7% of the total isolates), followed by Curvularia, Cephalosporium, and Humicola. Of nine isolated Aspergillus spp., A. humicola was the most dominant (37.4% of the total catch) and appeared at all polluted sites. Therefore, A. humicola was chosen to investigate its potential for heavy metals sorption from the contaminated waste water effluent. Four days old biomass pellets could sorb a large amount of heavy metals according to the following sequence: Zn>Cd>Cu>Ni ions. Agitation significantly increased Zn and Cd sorption, but not Cu and Ni. Heavy metals sorption took place at a wide pH range and particularly increased at alkaline pH (8-9). PMID- 12588105 TI - Quantitative [18F]fluorodopa/PET and histology of fetal mesencephalic dopaminergic grafts to the striatum of MPTP-poisoned minipigs. AB - The functional restoration of the dopamine innervation of striatum in MPTP poisoned Gottingen minipigs was assessed for 6 months following grafting of fetal pig mesencephalic neurons. Pigs were assigned to a normal control group and a MPTP-poisoned group, members of which received no further treatment, or which received bilateral grafts to the striatum of tissue blocks harvested from E28 fetal pig mesencephalon with and without immunosuppressive treatment after grafting, or with additional co-grafting with immortalized rat neural cells transfected to produce GDNF. In the baseline condition, and again at 3 and 6 months postsurgery, all animals were subjected to quantitative [18F]fluorodopa PET scans and testing for motor impairment. At the end of 6 months, tyrosine hydroxylase (TH)-containing neurons were counted in the grafts by stereological methods. The MPTP poisoning persistently reduced the magnitude of k3(D), the relative activity of DOPA decarboxylase in striatum, by 60%. Grafting restored the rate of [18F]fluorodopa decarboxylation to the normal range, and normalized the scores in motor function. The biochemical and functional recovery was associated with survival of approximately 100,000 TH-positive graft neurons in each hemisphere. Immunosuppression did not impart a greater recovery of [18F]fluorodopa uptake, nor were the number of TH-positive graft neurons or the volumes of the grafts increased in the immunosuppressed group. Contrary to expectation, co-grafting of transfected GDNF-expressing HiB5 cells, a rat-derived neural cell line, tended to impair the survival of the grafts with the lowest values for graft volumes, TH-positive cell numbers, behavioral scores, and relative DOPA decarboxylase activity. From the results we conclude that pig ventral mesencephalic allografts can restore functional dopamine innervation in adult MPTP-lesioned minipigs. PMID- 12588106 TI - Neovascularization induced by autologous bone marrow cell implantation in peripheral arterial disease. AB - Neovascularization has recently been used as a new treatment for severe ischemic disease. We tried to induce therapeutic neovascularization by autologous bone marrow cell implantation (BMCI) in eight selected patients with chronic peripheral arterial disease (PAD), in whom traditional treatments had failed. Improvement of subjective symptoms was seen in seven patients after treatment. Of three limbs with toe or finger ulceration, complete healing was achieved in two, while the other one became less severe after treatment. No relative toxicity was observed in any of the patients. BMCI might be a feasible treatment for selected patients with chronic PAD. PMID- 12588107 TI - Embryonic stem cells attenuate viral myocarditis in murine model. AB - We used mice to test our hypothesis that in response to viral invasion, stem cells may migrate into the heart and attenuate the effect of viral myocarditis. Male BALB/c mice were divided into three groups: mouse embryonic stem (ES) cell control, encephalomyocarditis virus (EMCV), and EMCV + ES cells. After administration of ES cells via tail vein, mice were immediately inoculated with EMCV. Mice were sacrificed at different days after EMCV inoculation. Mortality was recorded. Inflammatory cell infiltration and necrosis (major pathological changes of viral myocarditis) were evaluated by hematoxylin-eosin staining. ES cell migration and differentiation were identified by immunofluorescence. The survival rate in the EMCV + ES cell group (80%) was significantly increased (p < 0.05) over the EMCV-alone group (64%). Also, the incidence of inflammatory cell infiltration and myocardial lesions was lower in the EMCV + ES cell mice. Furthermore, the result of green fluorescent protein (GFP) and alpha-actinin analysis indicated that ES cells migrated into the heart and differentiated into myocytes after virus inoculation. In conclusion, ES cells significantly increased the survival of viral myocarditis mice and also decreased the necrosis and infiltration of inflammatory cells. These results demonstrated the ability of stem cells to mitigate the effects of viral infection on the heart and illustrated their potential therapeutic application to other mammalian species, including humans. PMID- 12588108 TI - Autotransplantation in mdx mice of mdx myoblasts genetically corrected by an HSV 1 amplicon vector. AB - Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder, characterized by a lack of dystrophin. To eliminate the need for immunosuppressive drugs, transplantation of genetically modified autologous myoblasts has been proposed as a possible therapy for this myopathy. An HSV-1 amplicon vector (HSVDGN), containing a 17.3-kb full-length MCK-driven mouse dystrophin cDNA, an eGFP gene, and a neomycin resistance gene driven by CMV or SV40 promoters, respectively, was constructed and used to transduce mdx primary myoblasts. The presence of the eGFP and neomycin resistance genes facilitated the evaluation of the initial transduction efficiency and the permanent transduction frequency. At low multiplicities of infection (MOI 1-5), the majority of myoblasts (60-90%) expressed GFP. The GFP-positive mdx myoblasts were sorted by FACS and selected with neomycin (300 microg/ml) for 2 weeks. Up to 2% of initially infected mdx myoblasts stably expressed the three transgenes without further selection at that time. These altered cells were grafted into the tibialis anterior muscles of 18 mdx mice. Some of the mice were immunosuppressed with FK506 due to the anticipation that eGFP and the product of neomycin resistance gene might be immunogenic. One month after transplantation, numerous muscle fibers expressing mouse dystrophin were detected by immunohistochemistry, in both immunosuppressed (10-50%) and nonimmunosuppressed (5-25%) mdx mice. Our results demonstrated the capability of permanently expressing a full-length dystrophin in dystrophic myoblasts with HSV-1 amplicon vector and raised the possibility of an eventual treatment of DMD based on the transplantation of genetically modified autologous myoblasts. PMID- 12588109 TI - Efficacy of the oxygen-charged static two-layer method for short-term pancreas preservation and islet isolation from nonhuman primate and human pancreata. AB - Previous reports indicate that the two-layer method (TLM) of human pancreas preservation is superior to University of Wisconsin solution (UW) when pancreata are preserved for extended periods (i.e., >24 h) prior to islet isolation. In this study, the efficacy of using the TLM for preserving pancreata for short periods (i.e., <13 h) was evaluated using both nonhuman primate and human pancreata preserved with a TLM kit precharged with oxygen. An oxygen precharged TLM (static TLM) was established and compared with the original TLM with continuous oxygen supply. For the static TLM, the perfluorochemical was fully oxygenated and the oxygen supply removed prior to pancreas preservation. In the primate model, pancreata were preserved by the static TLM, the original TLM, and UW for 5 h prior to islet isolation. In the human model, pancreata were preserved with the static TLM or the original TLM or UW for 4-13 h. Both primate and human pancreata were processed by intraductal collagenase injection and digestion followed by continuous density gradient purification to isolate islets. Islets were assessed for islet yield, purity, viability, and in vitro functionality. In the primate model, islet yield, viability, and in vitro functionality were significantly improved by both the static TLM and the original TLM with similar results. Postculture islet yields were 23,877 +/- 3619 IE/g in the static TLM, 21,895 +/- 3742 IE/g in the original TLM, and 6773 +/- 735 IE/g in UW. In the human model, both the static TLM and the original TLM significantly increased islet yield compared with UW with postculture islet yields of 2659 +/- 549 IE/g in the static TLM, 2244 +/- 557 IE/g in the original TLM, and 1293 +/- 451 IE/g in UW. Nonhuman primate and human pancreata stored in the static TLM, immediately upon procurement, yield isolated islets of a substantially higher quantity than when pancreata are stored in UW. Thus, the use of the static TLM should replace the use of UW for storage of pancreata during transport prior to islet isolation. PMID- 12588110 TI - Induction of chimerism in mice using human MHC class I-mismatched Hoechst 33342 side population donor stem cells. AB - A population of Hoechst 33342-stained cells, termed side population (SP) cells, can reconstitute the hematopoietic system of syngeneic mice. This study examined whether limiting numbers of SP cells can repopulate mice across a xenogeneic MHC class I barrier. SP cells were isolated from HLA.B7 and HLA.A2.1 transgenic mice by FACS and placed in colony assays or transplanted into irradiated C57BL/6 (B/6) recipients. SP cells contained few colony-forming cells when placed directly in culture. The number of GM-CFC and HPP-CFC increased up to 3000- and 300-fold, respectively, after 7 days in IL-3- and SCF-stimulated liquid culture. BMC derived GM-CFC increased up to only 12-fold and HPP-CFC decreased after 7 days in culture. HLA-B7 SP cells (2500-5000) were transplanted into lethal-irradiated B/6 mice. Two-color flow analysis, 4-6 weeks after transplantation, showed that HLA B7 expression in granulocyte-, macrophage-, and lymphocyte-specific lineages from reconstituted mice was similar to that in B7 transgenic mice. Secondary transplanted B/6 mice also showed a pattern of HLA-B7 expression similar to that in transgenic mice and were followed for longer than 16 weeks with stable chimerism. When HLA-A2.1 SP cells were transplanted into sublethally irradiated mice, 50% of the mice expressed HLA-A2 by PCR analysis in short-term repopulation studies. These data confirm that limiting numbers of SP cells can repopulate the major hematopoietic lineages in lethal and sublethally irradiated mice across a human MHC class I barrier. Therefore, SP cells may be useful for establishing mixed chimerism, which may induce immunologic nonresponsiveness to donor antigens in solid organ transplantation. PMID- 12588111 TI - Development of a novel cytomedical treatment that can protect entrapped cells from host humoral immunity. AB - Cell therapy is expected to relieve the shortage of donors needed for organ transplantation. When patients are treated with allogeneic or xenogeneic cells, it is necessary to develop a means by which to isolate administered cells from an immune attack by the host. We have developed "cytomedicine," which consists of functional cells entrapped in semipermeable polymer, and previously reported that alginate-poly-L-lysine-alginate microcapsules and agarose microbeads could protect the entrapped cells from injury by cellular immunity. However, their ability to isolate from humoral immunity was insufficient. It is well known that the complement system plays an essential role in rejection of transplanted cells by host humoral immunity. Therefore, the goal of the present study was to develop a novel cytomedical device containing a polymer capable of inactivating complement. In the screening of various polymers, polyvinyl sulfate (PVS) exhibited high anticomplement activity and low cytotoxicity. Murine pancreatic beta-cell line (MIN6 cell) entrapped in agarose microbeads containing PVS maintained viability and physiological insulin secretion, replying in response to glucose concentration, and resisted rabbit antisera in vitro. PVS inhibited hemolysis of sensitized sheep erythrocytes (EAs) and rabbit erythrocytes by the complement system. This result suggests that PVS inhibits both the classical and alternative complement pathways of the complement system. Next, the manner in which PVS exerts its effects on complement components was examined. PVS was found to inhibit generation of C4a and Ba generation in activation of the classical and alternative pathways, respectively. Moreover, when the EAC1 cells, which were carrying C1 on the EAs, treated with PVS were exposed to C1-deficient serum, hemolysis decreased in a PVS dose-dependent manner. These results suggest that PVS inhibits C1 in the classical pathway and C3 convertase formation in the alternative pathway. Therefore, PVS may be a useful polymer for developing an anticomplement device for cytomedical therapy. PMID- 12588112 TI - The immunoprotective effect of Sertoli cells coencapsulated with islet xenografts is not dependent upon Fas ligand expression. AB - Coencapsulation with Sertoli-enriched testicular cell fractions prolongs islet graft survival time compared with islet encapsulation alone in a highly discordant tilapia (fish)-to-mouse xenotransplantation model. Here we investigate whether Fas ligand (Fas-L) expression by testicular Sertoli cells is responsible for this additional protective effect. Sertoli-enriched testicular cell fractions (7 x 10(6) cells) harvested from either Fas-L-defective (group I) or Fas-L positive (group II) mice were coencapsulated in alginate gel spheres with fish islets and then transplanted into streptozotocin-diabetic Balb/c recipients. Group III mice received encapsulated islets without coencapsulated Sertoli cells. After transplantation, blood glucose levels were monitored three times per week. Mean graft survival times for the three groups were: group I = 35.6 +/- 10.2 days (n = 9), group II = 31.3 +/- 9.4 days (n = 7), and group III = 23.3 +/- 2.2 days (n = 6) (ANOVA, p = 0.043). Coencapsulation, regardless of the Fas-L status of the Sertoli cell donors, modestly prolonged graft survival. There was no significant difference between Fas-L-deficient and Fas-L-positive donors. Our results suggest that Fas/Fas-L interaction is not responsible for the additional protection afforded to encapsulated discordant islet xenografts by coencapsulation with Sertoli cells. PMID- 12588113 TI - Effects of carboxypeptidase E overexpression on insulin mRNA levels, regulated insulin secretion, and proinsulin processing of pituitary GH3 cells transfected with a furin-cleavable human proinsulin cDNA. AB - We recently developed two rat pituitary GH3 cell clones engineered to secrete human insulin (InsGH3). InsGH3 cells convert proinsulin into mature insulin, which is partially stored into a readily releasable pool of secretory granules. The efficiency of these processes, however, is relatively low in these cells, either in vitro or in vivo. This study was aimed at determining whether carboxypeptidase E (Cpe) overexpression can increase proinsulin processing and regulated secretion by InsGH3 clones. Indeed, in its membrane-bound form Cpe works as sorting receptor for the regulated secretory pathway of many hormones while, in its soluble form, Cpe takes part to the late step of insulin maturation. We obtained two Cpe-overexpressing cell lines from two different InsGH3 clones (InsGH3/C1 and C7). In the Cpe-overexpressing cell lines, derived from InsGH3 of clone 1 (InsGH3/C1-HACpe), in which the membrane-bound form of exogenous Cpe is accounted for by 90% of total Cpe immunoreactivity, we observed an increase in proinsulin gene expression, and in basal and stimulated insulin secretion compared with the original clone. In contrast, in the Cpe overexpressing cell line derived from InsGH3 of clone 7 (InsGH3/C7-HACpe), where the exogenous membrane-bound form was only 60% of total Cpe, we detected a decrease in basal insulin release and a modest, albeit significant, increase in intracellular proinsulin processing. In conclusion, Cpe overexpression can increase regulated insulin secretion and proinsulin processing in InsGH3 cells; however, such improvements appear quantitatively and qualitatively modest. PMID- 12588114 TI - Oxygen tension and blood flow in relation to revascularization in transplanted adult and fetal rat pancreatic islets. AB - We have previously recorded a decreased oxygen tension and blood flow in syngeneically transplanted rat pancreatic islets. The present study related measurements of oxygen tension and blood flow to the vascular density in such grafts implanted beneath the renal capsule. We also evaluated whether transplanted fetal islets are better revascularized than adult islets, and if the degree of revascularization is directly related to the islet vascular endothelial growth factor (VEGF) production. Tissue pO2 was measured using Clark microelectrodes, whereas islet graft blood flow was measured with laser-Doppler flowmetry. The vascular density of endogenous and transplanted islets was quantified in histological specimens stained with the lectin Bandeiraea simplicifolia (BS-1). Tissue pO2 in the transplanted adult and fetal islet grafts was similar and markedly lower than in the endogenous islets. The blood perfusion of both the adult and fetal islet grafts was 60-65% of that in the renal cortex. Administration of D-glucose did not affect tissue pO2 in either the endogenous or transplanted islets, nor graft blood perfusion. The number of capillaries found in the transplanted adult and fetal islets was similar and markedly lower than in endogenous islets. However, in the connective tissue stroma, which constituted approximately 20% of all islet grafts, the vascular density was higher than in the corresponding endocrine parts of these grafts. Incubated adult islets released higher amounts of VEGF than fetal islets. In conclusion, the previously described low oxygen tension of syngeneically transplanted adult rat islets is related to a low vascular density. Similar low oxygen tension and vascular density are seen in grafted fetal islets. The amount of VEGF production does not correlate to the degree of revascularization of the grafts. PMID- 12588115 TI - Evaluating the effect of serine proteases on collagenase activity during human islet isolation. AB - Inconsistencies in human islet yields after collagenase digestion have been attributed to the activation of endogenous enzymes of the donor pancreas. It has been suggested that pancreatic serine proteases contribute to the proteolysis of collagenase. This study defined the effects of endogenous enzymes within the pancreas on pancreas dissociation during collagenase digestion. Levels of collagenase activity from samples taken throughout several steps in islet isolation procedures, both with and without the addition of the serine protease inhibitor Pefabloc, were determined by a spectrophotometric assay using N-[3-(2 furyl)acryloyl]-Leu-Gly-Pro-Ala as the substrate. Results clearly demonstrated that the level of collagenase activity remains stable throughout the isolation procedure despite differences in the donor factors from several cadaveric donor pancreases. This was further demonstrated by observing no difference in activity levels after incubating commercial collagenase preparations with serine proteases and analyzing by means of collagenase activity and SDS-PAGE. These data show that the presence of serine proteases does not affect the level of collagenase activity; however, they likely damage the islet cells upon prolonged digestion of the pancreatic tissue. Further efforts at examining exogenous and endogenous enzyme levels may result in the development of an enzyme cocktail that is both stable and effective for digesting the human pancreas while preserving islet function and viability. PMID- 12588116 TI - The morphology of islets within the porcine donor pancreas determines the isolation result: successful isolation of pancreatic islets can now be achieved from young market pigs. AB - Clinical islet allotransplantation has become an increasingly efficient "routine" therapy in recent years. Shortage of human donor organs leads to porcine pancreatic islets as a potential source for islet xenotransplantation. Yet it is still very difficult to isolate sufficient numbers of intact porcine islets, particularly from young market pigs. In the following study islets were successfully isolated from retired breeders [4806 +/- 720 islet equivalents per gram organ (IEQ/g); n = 25; 2-3 years old; RB] and also from young hybrid pigs [2868 +/- 260 IEQ/g; n = 65; 4-6 months old; HY] using LiberasePI and a modified version of Ricordi's digestion-filtration technique. As expected, isolations from RB showed significantly better results (p < 0.002). A retrospective histological analysis of almost all donor pancreases showed that the majority of organs from RB (80%) contained mainly large islets (diameter > 200 microm), in contrast to only 35% of all pancreases from HY. Remarkably, the islet size in situ, regardless whether detected in RB or HY, strongly determined the isolation result. A donor organ with predominantly large islets resulted in significantly higher numbers of IEQs compared with a donor organ with predominantly small islets [RB(Large Islets): 5680 +/- 3,318 IEQ/g (n= 20); RB(Small Islets): 1353 +/ 427 IEQ/g (n = 5); p < 0.02]. In addition, isolation results were strongly influenced by the quality of the LiberasePI batch, and therefore single batch testing is invariably required. Purification was performed using Ficoll or OptiPrep density gradient centrifugation manually or in the COBE cell processor. Although islet purity was highest when OptiPrep was used, final islet yields did not differ between the different purification methods. Our study demonstrates that islet size in situ is an extremely critical parameter for highly successful islet isolation; consequently, we are now performing a morphological screening of each donor organ prior to the isolation process. Under these conditions highly successful isolations can reliably be performed even from young market pigs. PMID- 12588117 TI - Infrared activity of symmetric bridging ligand modes in pyrazine-bridged hexaruthenium mixed-valence clusters. AB - A fully symmetric (A(g)) vibrational mode of pyrazine is observed in the infrared spectrum of four pyrazine-bridged hexaruthenium mixed-valence complexes with varying degrees of electronic coupling between clusters. Deuteration of the bridging pyrazine ligand and the accompanying shift in frequency confirm the assignment of this mode. Previous observation of infrared line coalescence in the carbonyl stretching region assigns all of these complexes to Robin-Day class II (partial localization of charge) on the picosecond time scale. The infrared activity of the fully symmetric bridging ligand mode could provide a complementary assignment of these complexes to class II on a faster, femtosecond time scale. However, the extinction coefficient for this band is much greater than that observed in similar asymmetric, non-mixed-valence complexes and suggests that its strong IR activity is due to vibronic enhancement rather than electronic asymmetry. PMID- 12588118 TI - A mono-diazenide complex from perrhenate: toward a new core for rhenium radiopharmaceuticals. AB - A new method for the synthesis of low to intermediate oxidation state rhenium complexes containing a bifunctional ligand has been developed. Reaction of [ReO(4)](-) with substituted phenylhydrazines and triphenylphosphine in acetonitrile in the presence of HCl allows the isolation of [ReCl(2)(NNC(6)H(4)-4 R)(NCCH(3))(PPh(3))(2)] (where R = OCH(3), Cl, or CO(2)CH(3)). The substituted hydrazine acts as both a reductant and source of a monodentate diazenide ligand. The compounds have all been characterized in the solid state by X-ray crystallography and in the solution state by NMR, electrospray mass spectrometry, and HPLC. Cyclic voltammetry measurements show that the mono-diazenide complexes undergo a reversible oxidation. PMID- 12588119 TI - Perfluoroalkyl-substituted triazapentadienes and their metal complexes. AB - Triazapentadienides, C(3)F(7)-C(=NR)-N=C(NHR)-C(3)F(7), result from the reaction of primary amines RNH(2) with the fluorinated imine C(3)F(7)-CF=N-C(4)F(9). The aniline derivative (R = Ph) is a weak monoprotic acid in dmso. Its conjugate base exhibits an extensive coordination chemistry. It acts as a bidentate ligand toward the molecular fragments Pd(C(3)H(5)), Rh(c-C(8)H(12)), Ir(c-C(8)H(12)), and Rh(CO)(2). The chelates [C(3)F(7)-C(NPh)-N-C(NPh)-C(3)F(7)](2)M, M = Mg, Mn, Fe, Co, Ni, Cu, Zn, and Pd, were prepared. In the crystallographically characterized Co complex, the metal is 3d(7), S = (3)/(2) and tetrahedrally coordinated. Spin densities at carbon in the C(6)H(5) and C(3)F(7) groups were estimated from the (1)H and (19)F contact shifts. Spin delocalization onto phenyl sp(2) carbons is approximately 10 times greater than onto the fluorinated sp(3) carbons. PMID- 12588120 TI - Photochromic isomerization of a dinuclear molybdenum complex with ethylene-1,2 dithiolate and disulfur ligands: x-ray structures of the two isomers. AB - A photochromic complex with disulfur and dimethyl-ethylene-1,2-dithiolate ligands, [Mo(2)(mu-S(2))(mu-S(2)C(2)Me(2))(2)(S(2)C(2)Me(2))(2)] (3), was synthesized and characterized. Photoirradiation of 3 with visible light resulted in the formation of the isomer (3'). The electronic spectrum of 3' has a new intense peak in the near infrared region, and in the dark, the spectrum returns to that of 3. X-ray structural analyses of 3.C(6)H(6) and 3' revealed a large conformational change of the bridging dithiolate ligands: the two ligands in 3' come very close to each other compared to those in 3.C(6)H(6). Crystal data: 3.C(6)H(6), monoclinic, space group C2/c, a = 15.193(4) A, b = 14.287(3) A, c = 14.685(4) A, beta = 105.30(1) degrees, V = 3074(1) A(3), Z = 4; 3', monoclinic, space group C2/c, a = 21.5400(8) A, b = 9.5232(5) A, c = 13.9828(2) A, beta = 118.924(1) degrees, V = 2510.5(2) A(3), Z = 4. (1)H NMR spectra of 3 (3.06, 3.05, 1.66, and 1.31 ppm) and 3' (2.90, 2.75, 2.14, and 1.97 ppm) are also reported: each spectrum has four signals due to methyl groups, which accords well with the fact that each of the molecules, 3.C(6)H(6) and 3', has a crystallographic 2-fold axis. PMID- 12588122 TI - Unprecedented reactivity of the bridged borylene complexes [mu-BCl((eta(5) C(5)H(4)Me)Mn(CO)(2))(2)] toward pyridine. AB - The reactivity of the bridged chloroborylene complex [mu-BCl((eta(5) C(5)H(4)Me)Mn(CO)(2))(2)] (1) toward pyridine was investigated under various conditions. In the presence of protic reagents such as H[Co(CO)(4)] or H[BF(4)], the formation of the aminoborylene complex [1-(mu-B)-4-H (NC(5)H(5))((C(5)H(4)Me)Mn(CO)(2))(2)] (2) was observed. Compound 2 represents the product of an unprecedented formal 1,4-hydroboration of pyridine. Corresponding reactions of 1 with pyridine and Tl[PF(6)] afforded 2 in similar yields, thus providing evidence that the abstraction of the boron bound chloride initiates the observed reaction. Complex 2 was fully characterized in solution and in the crystal. PMID- 12588121 TI - Formation of a stable cyano-bridged dinuclear iron cluster following oxidation of the superoxide reductases from Treponema pallidum and Desulfovibrio vulgaris with K(3)Fe(CN)(6). AB - Superoxide reductases catalyze the monovalent reduction of superoxide anion to hydrogen peroxide. Spectroscopic evidence for the formation of a dinuclear cyano bridged adduct after K(3)Fe(CN)(6) oxidation of the superoxide reductases neelaredoxin from Treponema pallidum and desulfoferrodoxin from Desulfovibrio vulgaris was reported. Oxidation with K(3)Fe(CN)(6) reveals a band in the near-IR with lambda(max) at 1020 nm, coupled with an increase of the iron content by almost 2-fold. Fourier transform infrared spectroscopy provided additional evidence with CN-stretching vibrations at 2095, 2025-2030, and 2047 cm(-)(1), assigned to a ferrocyanide adduct of the enzyme. Interestingly, the low temperature electronic paramagnetic resonance (EPR) spectra of oxidized TpNlr reveal at least three different species indicating structural heterogeneity in the coordination environment of the active site Fe ion. Given the likely 6 coordinate geometry of the active site Fe(3+) ion in the ferrocyanide adduct, we propose that the rhombic EPR species can serve as a model of a hexacoordinate form of the active site. PMID- 12588123 TI - Photoluminescent metal-organic polymer constructed from trimetallic clusters and mixed carboxylates. AB - The solvothermal reaction of zinc acetate dihydrate with a mixture of benzene-1,4 dicarboxylic acid (H(2)BDC) and benzene-1,3,5-tricarboxylic acid (H(3)BTC) in a solution containing N,N'-dimethylformamide (DMF), absolute ethanol, and chlorobenzene gave rise to a metal-organic polymer, Zn(3).BDC.2BTC.2NH(CH(3))(2).2NH(2)(CH(3))(2). The structure of this polymer possesses a unique three-dimensional framework with tri-zinc clusters, and BDC and BTC units colinking the clusters. Moreover, this metal-organic polymer exhibits strong photoluminescence at room temperature, and the main emission band is at about 430 nm (lambda(ex) = 325 nm). Crystal data for this compound (C(17)H(20)N(2)O(8)Zn(1.5)): monoclinic, space group P2(1)/n, cell dimensions a = 11.6171(3) A, b = 14.2456(4) A, c = 12.6426(3) A, beta = 107.030(2) degrees, V = 2000.51(9) A(3), and Z = 4. PMID- 12588124 TI - Crystal growth of novel osmium-containing triple perovskites. AB - Single crystals of two new osmium-containing triple perovskites, Ba(3)LiOs(2)O(9) and Ba(3)NaOs(2)O(9), were grown from reactive molten hydroxide fluxes in sealed silver tubes. They crystallize in the space group P6(3)/mmc with lattice parameters of a = 5.8025(1) A, c = 14.1468(4) A for Ba(3)LiOs(2)O(9) and a = 5.8858(1) A, c = 14.3451(5) A for Ba(3)NaOs(2)O(9). The magnetic susceptibility of these osmates indicates significant Os-Os coupling within the octahedra pair. PMID- 12588125 TI - Synthesis and characterization of rhenium(V) oxo complexes with N-[N-(3 diphenylphosphinopropionyl)glycyl]cysteine methyl ester. X-ray crystal structure of (ReO[Ph(2)P(CH(2))(2)C(O)-Gly-Cys-OMe(P,N,N,S)]). AB - The PN(2)S chelate N-[N-(3-diphenylphosphinopropionyl)glycyl]-S-tritylcysteine methyl ester [PN(2)S(Trt)-OMe] was synthesized and reacted with ReOCl(3)(PPh(3))(2) and Ph(4)P[ReOCl(4)]. The reactions of both tritylated and detritylated ligands with Re(V)O precursors gave two diastereomers, 9a and 9b, of the ReO(PN(2)S-OMe) complex. The two isomers, produced in a 1:1 molar ratio, are stable and do not interconvert. They were separated by reverse-phase HPLC and characterized by NMR, FT-IR, and UV-visible spectroscopy and electrospray mass spectrometry. X-ray analysis established for 9a the presence in the solid of the syn isomer. Compound 9a, C(21)H(23)N(2)O(5)PSRe, crystallized from warm acetonitrile in the triclinic space group Ponemacr;, a = 9.828(2) A, b = 11.163(2) A, c = 11.641(2) A, alpha = 106.48(3) degrees, beta = 109.06(3) degrees, gamma = 102.81(3) degrees, V = 1085.7(4) A(3), Z = 2. The PN(2)S coordination set is in the equatorial plane, and the complex shows a distorted square pyramidal coordination. The anti configuration assigned to 9b is consistent with all the available physicochemical data. Follow-up of the reaction of the detritylated ligand with Ph(4)P[ReOCl(4)] in ethanol or acetonitrile indicated that the phosphorus atom of the chelate binds first to the metal and that this bond acts as the driving force for coordination. PMID- 12588126 TI - Water displacement by cyanogold complexes in binuclear nickel(II) compounds based on bridging oxalate. Synthesis, structural diversity, magnetic properties, and DFT calculations. AB - Several cyanogold complexes react with the binuclear nickel complex [(Ni(dien)(H(2)O))(2)(mu-ox)](PF(6))(2).2H(2)O to give the compounds [(Ni(dien)(H(2)O))(2)(mu-ox)]Br(2) (1), [(Ni(dien)(Au(CN)(2)))(2)(mu-ox)] (2), and [(Ni(dien))(2)(mu-ox)(mu-Au(CN)(4))](PF(6)) (3) (dien, diethilenetriamine; ox, oxalate). In the case of compounds 2 and 3, water displacement by the corresponding cyanogold complex takes place, whereas compound 1 is formed by a substitution of the anion. The crystal structures of compounds 1 and 2 present a 2D arrangement where the layers are connected by van der Waals forces (1) or N H.Ntbd1;C hydrogen bonds (2), where each binuclear complex is hydrogen bonded to its neighbors, whereas compound 3 presents a novel structure where the tetracyanoaurate acts as a bridging ligand to give a polymeric compound. Magnetic studies of these compounds reveal an antiferromagnetic behavior. Finally, density functional theory (DFT) calculations have been performed on isolated models of compounds 2 and 3 in order to gain some insight about the different behavior of the [Au(CN)(2)](-) and [Au(CN)(4)](-) groups as ligands and proton acceptors in hydrogen bonds. PMID- 12588127 TI - Synthesis and x-ray crystal structure of [(THF)Zn(O(2)(OH)SiR)](4) (R = (2,6-i Pr(2)C(6)H(3))N(SiMe(3))): enroute to larger aggregates. AB - Reaction of aminosilanetriol RSi(OH)(3) (1) (R = (2,6-i-Pr(2)C(6)H(3))N(SiMe(3))) with diethyl zinc at room temperature in 1:1 stoichiometric ratio affords [(THF)Zn(O(2)(OH)SiR)](4) (2) (R = (2,6-i-Pr(2)C(6)H(3))N(SiMe(3))) in good yield. The single-crystal X-ray diffraction studies reveal that 2 is monoclinic, P2(1), with a = 17.117(3) A, b = 16.692(5) A, c = 17.399(4) A, alpha = gamma = 90 degrees, beta = 91.45(7) degrees, and Z = 2. The molecular structure of 2 contains two puckered eight-membered Zn(2)Si(2)O(4) rings, which are connected by the Zn-O bonds and form two planar four-membered Zn(2)O(2) rings. Compound 2 contains an unreacted hydroxyl group on each silicon atom, and hence, we carried out the reactions of 2 with dimethylzinc and methyllithium to form [Zn(4)(THF)(4)(MeZn)(4)(O(3)SiR)(4)] (3) (R = (2,6-i-Pr(2)C(6)H(3))N(SiMe(3))) and [(L)ZnLi(O(3)SiR)](4) (4) (L = 1,4-(Me(2)N)(2)C(6)H(4), R = (2,6-i Pr(2)C(6)H(3))N(SiMe(3))), respectively. This suggested that 2 could be an intermediate product formed during the synthesis of 3 and 4. PMID- 12588128 TI - A complete family of isostructural cluster compounds with cubane-like M(3)S(4)M' cores (M = Mo, W; M' = Ni, Pd, Pt): comparative crystallography and electrochemistry. AB - By reaction of the geometrically incomplete cubane-like clusters [(eta(5) Cp')(3)Mo(3)S(4))][pts] and [(eta(5)-Cp')(3)W(3)S(4)][pts] (Cp' = methylcyclopentadienyl; pts = p-toluenesulfonate) with group 10 alkene complexes, three new heterobimetallic clusters with cubane-like cluster cores were isolated: [(eta(5)-Cp')(3)W(3)S(4)M'(PPh(3))][pts] ([5][pts], M' = Pd; [6][pts], M' = Pt); [(eta(5)-Cp')(3)Mo(3)S(4)Ni(AsPh(3))][pts] ([7][pts]). The compounds [5][pts] [7][pts] are completing the extensive series of clusters [(eta(5) Cp')(3)M(3)S(4)M'(EPh(3))][pts] (M = Mo, W; M' = Ni, Pd, Pt; E = P, As) which allows the consequences of replacing a single type of atom on structural and NMR and UV/vis spectroscopic as well as electrochemical properties to be determined. Single-crystal X-ray structure determinations of [5][pts]-[7][pts] revealed that [5][pts] was not isomorphous to the other members of the series [(eta(5) Cp')(3)M(3)S(4)M'(EPh(3))][pts] due to distinctly different cell parameters, which in the molecular structure of [5](+) is reflected in a slightly different orientation of the PPh(3) ligand. Electrochemical measurements on the series showed that the Mo-based clusters were more difficult to oxidize than their W based analogues. The Pd-containing clusters underwent two-electron oxidation processes, whereas the Ni- and Pt-containing clusters underwent two separated one electron oxidation processes. PMID- 12588129 TI - A high-pressure iron K-edge x-ray absorption spectral study of the spin-state crossover in (Fe[HC(3,5-(CH(3))(2)pz)(3)](2))I(2) and (Fe[HC(3,5 (CH(3))(2)pz)(3)](2))(BF(4))(2). AB - The room temperature iron K-edge X-ray absorption near edge structure spectra of (Fe[HC(3,5-(CH(3))(2)pz)(3)](2))I(2) and (Fe[HC(3,5 (CH(3))(2)pz)(3)](2))(BF(4))(2) have been measured between ambient and 88 and 94 kbar, respectively, in an opposed diamond anvil cell. The iron(II) in (Fe[HC(3,5 (CH(3))(2)pz)(3)](2))I(2)undergoes the expected gradual spin-state crossover from the high-spin state to the low-spin state with increasing pressure. In contrast, the iron(II) in (Fe[HC(3,5-(CH(3))(2)pz)(3)](2))(BF(4))(2) remains high-spin between ambient and 78 kbar and is only transformed to the low-spin state at an applied pressure of between 78 and 94 kbar. No visible change is observed in the preedge peak in the spectra of (Fe[HC(3,5-(CH(3))(2)pz)(3)](2))I(2) with increasing pressure, whereas the preedge peak in the spectra of ((e[HC(3,5 (CH(3))(2)pz)(3)](2))(BF(4))(2) changes as expected for a high-spin to low-spin crossover with increasing pressure. The difference in the spin-state crossover behavior of these two complexes is likely related to the unusual behavior of (Fe[HC(3,5-(CH(3))(2)pz)(3)](2))(BF(4))(2) upon cooling. PMID- 12588130 TI - Structural and physical properties of the ferromagnetic tris-dithiooxalato compounds, A[M(II)Cr(III)(C(2)S(2)O(2))(3)], with A(+) = N(n C(n)()H(2)(n)(+1))(4)(+) (n = 3-5) and P(C(6)H(5))(4)(+) and M(II) = Mn, Fe, Co, and Ni. AB - The structural and magnetic properties of the tris-dithiooxalato salts, A[M(II)Cr(C(2)S(2)O(2))(3)], have been investigated with A(+) = PPh(4)(+), N(n C(n)()H(2)(n)()(+1))(4)(+), with n = 3-5, where M(II) is Mn, Fe, Co, and Ni. With the exception of A[MnCr(C(2)S(2)O(2))(3)], all the salts are ferromagnets with Curie temperatures, T(c), between 5 and 16 K. In contrast to the corresponding oxalates which are ferromagnetic, the A[MnCr(C(2)S(2)O(2))(3)] compounds are paramagnetic above 2 K. Powder neutron diffraction studies of d(20) PPh(4)[FeCr(C(2)S(2)O(2))(3)] indicate that no structural phase transitions occur between 2.4 and 285 K and that the coefficient of linear expansion is four times larger for the c-axis than for the a-axis. The crystal structure refined from powder neutron diffraction data confirms the honeycomb layer arrangement observed in the related bimetallic tris-oxalate salts. The Mossbauer spectra reveal that the iron(II) in PPh(4)[FeCr(C(2)S(2)O(2))(3)] is coordinated mainly to six oxygen atoms of the dithiooxalato ligand but with a minor component of sulfur coordination that increases with aging of the sample; the iron(II) is high-spin in both cases. Powder neutron diffraction profiles of d(20) PPh(4)[FeCr(C(2)S(2)O(2))(3)] below T(c) show magnetic intensity with a q = 0 propagation vector, confirming the presence of ferromagnetic order. PMID- 12588131 TI - Ground-state stability and rotational activation parameters for individual rotamers of (R,S,S,R)-(N,N'-dimethyl-2,3-diaminobutane)PtG(2) complexes (G = 9 EtG, 3'-GMP, and 5'-GMP). AB - Rate constants for guanine rotation about the Pt-N7 bond in (R,S,S,R) Me(2)DABPtG(2) complexes (Me(2)DAB = N,N'-dimethyl-2,3-diaminobutane; G = 9-EtG, 3'-GMP, and 5'-GMP) were evaluated from line-shape analysis of H8 resonances. Three diastereomers, two in head-to-tail (DeltaHT and LambdaHT) and one in head to-head (HH) conformations, exist in equilibrium in solution. The two guanines are equivalent in DeltaHT and LambdaHT conformers and nonequivalent in the HH form; therefore, four rate constants (k(Delta)(HT), k(Lambda)(HT), k(HH)()s, and k(HH)()d; sub-subscripts s and d stand for H8-shielded and -deshielded guanine, respectively) were evaluated. Activation parameters (DeltaH and DeltaS) were evaluated from the rate constant dependence on temperature. High values of DeltaH (78-93 kJ mol(-)(1)) and DeltaS (51-71 J K(-)(1) mol(-)(1)) were found for G rotation in the preferred DeltaHT rotamer having the six-membered ring of each guanine more canted toward the cis-G and a favorable dipole-dipole internucleotide interaction. Lower values of DeltaH() (64-76 kJ mol(-)(1)) and very small values of DeltaS() (-7-11 J K(-)(1) mol(-)(1)) were found for G rotation in the less favorable LambdaHT rotamer, indicating that the ground-state entropy of this rotamer is close to that of the activated complex and the ground state enthalpy closer to that of the activated complex than for the DeltaHT rotamer. For the two guanines in the HH rotamer there is no large difference in activation parameters. In general DeltaH falls in the range 66-84 kJ mol(-)(1) (rather close to the values for the LambdaHT rotamer) and DeltaS in the range 14 41 J K(-)(1) mol(-)(1). The equilibrium constant between HT and HH rotamers was also evaluated together with the corresponding thermodynamic parameters (DeltaH and DeltaS). It is found that the low enthalpy is the major stabilizing factor for DeltaHT as compared to HH, while the entropy factor would favor the latter rotamer. In contrast the greater entropy is the stabilizing factor for the LambdaHT rotamer (the second most abundant conformer for 9-EtG and 3'-GMP) over the HH rotamer. In the latter case the enthalpy would favor the HH rotamer. In the case of the 5'-GMP derivative the greater entropy of the LambdaHT rotamer is not such to compensate for the lower enthalpy of the HH rotamer, and the latter remains the second most abundant rotamer. This investigation has allowed for the first time the enthalpic and entropic contributions favoring different rotamers to be distinguished. PMID- 12588132 TI - 31P NMR spectroscopy as a powerful tool for the determination of enantiomeric excess and absolute configurations of alpha-amino acids. AB - An easy method for the determination of the enantiomeric excess (ee) of mixtures of alpha-amino acids, and also for the elucidation of the absolute configuration of each component of the mixture, is reported. The method is based on the formation of diastereoisomers by reaction of the enantiomerically pure acetylacetonate derivative [Pd(acac-O,O')(P(2)-dach)]ClO(4) (4) [P(2)-dach = (1R,2R)-C(6)H(10)(NHPPh(2))(2)] with d,l-mixtures of alpha-amino acids AaH (Pd:AaH = 1:1 molar ratio, refluxing MeOH). The reaction occurs with protonation of the acac ligand and N,O-coordination of the amino acidate group, giving the corresponding [Pd(Aa-N,O)(P(2)-dach)]ClO(4) complexes l-5 and d-6. The composition of these mixtures of amino acidate complexes was analyzed by integration of the corresponding peaks (four doublets, two for each diastereomer) in their (31)P((1)H) NMR spectra. A series of 14 alpha-amino acids was studied (a, alanine; b, 2-aminobutyric acid; c, valine; d, phenylalanine; e, proline; f, leucine; g, isoleucine; h, norleucine; i, serine; j, threonine; k, methionine; l, aspartic acid; m, glutamine; n, cysteine), and excellent agreement between the expected values of ee and those obtained from integration of the (31)P((1)H) NMR spectra was obtained. Moreover, the position of the signals of each isomer is diagnostic, in such a way that the outer doublets are always due to the l derivatives 5a-l, while the inner ones are due to the d-derivatives 6a-l, allowing the assignation of absolute configurations to each isomer in the mixture. PMID- 12588133 TI - Aqueous synthesis of derivatized cyclopentadienyl complexes of technetium and rhenium directed toward radiopharmaceutical application. AB - Half-sandwich complexes of the type [(RCOCp)M(CO)(3)] with M = Re and (99(m))Tc were synthesized from [M(OH(2))(3)(CO)(3)](+) in water. The R group can be an organic residue or a receptor binding biomolecule with a spacer to cyclopentadienyl (Cp). This provides a general route to Cp complexes of technetium without the need for starting from [TcBr(CO)(5)]. The X-ray structure of [(C(6)H(5)CH(2)COC(5)H(4))Tc(CO)(3)] has been elucidated. The compound crystallizes in the monoclinic space group P2(1)/c with a = 16.1454(9), b = 7.6300(6), and c = 12.3922(7) A and beta = 107.792(6) degrees. We have chosen a serotonergic receptor ligand (WAY) as an example for the derivatization of Cp with a bioactive molecule. WAY is linked to Cp by an aliphatic chain of variable length. The half-sandwich complexes were prepared from water and organic solvents. The structure of [(WAY4-Cp)Re(CO)(3)] could be elucidated. The compound crystallizes in the monoclinic space group P2(1)/c with a = 15.7112(6), b = 6.8775(3), and c = 25.5217(12) A and beta = 103.778(5) degrees. Quantification of inhibition constants gave a clear structure-activity relationship. A single methylene group between the receptor binding site and the half-sandwich complex gave an IC(50) of 217 nM for HT(1A), whereas a butylene linker resulted in retention of the inhibition constant with an IC(50) of 6 nM with respect to underivatized WAY. For use as radiopharmaceuticals, the compounds have also been prepared with (99m)Tc in quantitative yield. PMID- 12588135 TI - Electrochemical and spectroelectrochemical studies on UO(2)(saloph)L (saloph = N,N'-disalicylidene-o-phenylenediaminate, L=dimethyl sulfoxide or N,N dimethylformamide). AB - To examine properties of pentavalent uranium, U(V), we have carried out electrochemical and spectroelectrochemical studies on UO(2)(saloph)L [saloph = N,N'-disalicylidene-o-phenylenediaminate, L = dimethyl sulfoxide (DMSO) or N,N dimethylformamide (DMF)]. The electrochemical reactions of UO(2)(saloph)L complexes in L were found to occur quasireversibly. The reduction processes of UO(2)(saloph)L complexes were followed spectroelectrochemically by using an optical transparent thin layer electrode cell. It was found that the absorption spectra measured at the applied potentials from 0 to -1.650 V versus ferrocene/ferrocenium ion redox couple (Fc/Fc(+)) for UO(2)(saloph)DMSO in DMSO have clear isosbestic points and that the evaluated electron stoichiometry equals 1.08. These results indicate that the reduction product of UO(2)(saloph)DMSO is [U(V)O(2)(saloph)DMSO](-), which is considerably stable in DMSO. Furthermore, it was clarified that the absorption spectrum of the [U(V)O(2)(saloph)DMSO](-) complex has a very small molar absorptivity in the visible region and characteristic absorption bands due to the 5f(1) orbital at around 750 and 900 nm. For UO(2)(saloph)DMF in DMF, the clear isosbestic points were not observed in the similar spectral changes. It is proposed that the UO(2)(saloph)DMF complex is reduced to [U(V)O(2)(saloph)DMF](-) accompanied by the dissociation of DMF as a successive reaction. The formal redox potentials of UO(2)(saloph)L in L (E(0), vs Fc/Fc(+)) for U(VI)/U(V) couple were determined to be -1.550 V for L = DMSO and 1.626 V for L = DMF. PMID- 12588134 TI - A new zinc(II) fluorophore 2-(9-anthrylmethylamino)ethyl-appended 1,4,7,10 tetraazacyclododecane. AB - A new 2-(9-anthrylmethylamino)ethyl-appended cyclen, L(3) (1-(2-(9 anthrylmethylamino)ethyl)-1,4,7,10-tetraazacyclododecane) (cyclen = 1,4,7,10 tetraazacyclododecane), was synthesized and characterized for a new Zn(2+) chelation-enhanced fluorophore, in comparison with previously reported 9 anthrylmethylcyclen L(1) (1-(9-anthrylmethyl)-1,4,7,10-tetraazacyclododecane) and dansylamide cyclen L(2). L(3) showed protonation constants log K(a)(i)() of 10.57 +/- 0.02, 9.10 +/- 0.02, 7.15 +/- 0.02, <2, and <2. The log K(a3) value of 7.15 was assigned to the pendant 2-(9-anthrylmethylamino)ethyl on the basis of the pH dependent (1)H NMR and fluorescence spectroscopic measurements. The potentiometric pH titration study indicated extremely stable 1:1 Zn(2+)-L(3) complexation with a stability constant log K(s)(ZnL(3)) (where K(s)(ZnL(3)) = [ZnL(3)]/[Zn(2+)][L(3)] (M(-)(1))) of 17.6 at 25 degrees C with I = 0.1 (NaNO(3)), which is translated into the much smaller apparent dissociation constant K(d) (=[Zn(2+)](free)[L(3)](free)/[ZnL(3)]) of 2 x 10(-)(11) M with respect to 5 x 10(-)(8) M for L(1) at pH 7.4. The quantum yield (Phi = 0.14) in the fluorescent emission of L(3) increased to Phi = 0.44 upon complexation with zinc(II) ion at pH 7.4 (excitation at 368 nm). The fluorescence of 5 microM L(3) at pH 7.4 linearly increased with a 0.1-5 microM concentration of zinc(II). By comparison, the fluorescent emission of the free ligand L(1) decreased upon binding to Zn(2+) (from Phi = 0.27 to Phi = 0.19) at pH 7.4 (excitation at 368 nm). The Zn(2+) complexation with L(3) occurred more rapidly (the second-order rate constant k(2) is 4.6 x 10(2) M(-)(1) s(-)(1)) at pH 7.4 than that with L(1) (k(2) = 5.6 x 10 M(-)(1) s(-)(1)) and L(2) (k(2) = 1.4 x 10(2) M(-)(1) s(-)(1)). With an additionally inserted ethylamine in the pendant group, the macrocyclic ligand L(3) is a more effective and practical zinc(II) fluorophore than L(1). PMID- 12588137 TI - Dinuclear cyano complexes of cobalt(III) and Iron(III) containing noninnocent 1,2,4,5-tetrakis(2-pyridinecarboxamido)benzene bridging ligands. AB - Two new dinucleating ligands 1,2,4,5-tetrakis(2-pyridinecarboxamido)benzene, H(4)(tpb), and 1,2,4,5-tetrakis(4-tert-butyl-2-pyridinecarboxamido)benzene, H(4)(tbpb), have been synthesized, and the following dinuclear cyano complexes of cobalt(III) and iron(III) have been isolated: Na(2)[Co(III)(2)(tpb)(CN)(4)] (1); [N(n-Bu)(4)](2)[Co(III)(2)(tbpb)(CN)(4)] (2); [Co(III)(2)(tbpb(ox2))(CN)(4)] (3); [N(n-Bu)(4)](2)[Fe(III)(2)(tpb)(N(3))(4)] (4); [N(n Bu)(4)](2)[Fe(III)(2)(tpb)(CN)(4)] (5); [N(n-Bu)(4)](2)[Fe(III)(2)(tbpb)(CN)(4)] (6). Complexes 2-4 and 6 have been structurally characterized by X-ray crystallography at 100 K. From electrochemical and spectroscopic (UV-vis, IR, EPR, Mossbauer) and magnetochemical investigations it is established that the coordinated central 1,2,4,5-tetraamidobenzene entity in the cyano complexes can be oxidized in two successive one-electron steps yielding paramagnetic (tbpb(ox1))(3)(-) and diamagnetic (tbpb(ox2))(2)(-) anions. Thus, complex 6 exists in five characterized oxidation levels: [Fe(III)(2)(tbpb(ox2))(CN)(4)](0) (S = 0); [Fe(III)(2)(tbpb(ox1))(CN)(4)](-) (S = (1)/(2)); [Fe(III)(2)(tbpb)(CN)(4)](2)(-) (S = 0); [Fe(III)Fe(II)(tbpb)(CN)(4)](3)(-) (S = (1)/(2)); [Fe(II)(2)(tbpb)(CN)(4)](4)(-) (S = 0). The iron(II) and (III) ions are always low-spin configurated. The electronic structure of the paramagnetic iron(III) ions and the exchange interaction of the three-spin system [Fe(III)(2)(tbpb(ox1))(CN)(4)](-) are characterized in detail. Similarly, for 2 three oxidation levels have been identified and fully characterized: [Co(III)(2)(tbpb)(CN)(4)](2)(-) (S = 0); [Co(III)(2)(tbpb(ox1))(CN)(4)](-) (S = (1)/(2)); [Co(III)(2)(tbpb(ox2))(CN)(4)](0). The crystal structures of 2 and 3 clearly show that the two electron oxidation of 2 yielding 3 affects only the central tetraamidobenzene part of the ligand. PMID- 12588136 TI - 5' modification of duplex DNA with a ruthenium electron donor-acceptor pair using solid-phase DNA synthesis. AB - Incorporation of metalated nucleosides into DNA through covalent modification is crucial to measurement of thermal electron-transfer rates and the dependence of these rates with structure, distance, and position. Here, we report the first synthesis of an electron donor-acceptor pair of 5' metallonucleosides and their subsequent incorporation into oligonucleotides using solid-phase DNA synthesis techniques. Large-scale syntheses of metal-containing oligonucleotides are achieved using 5' modified phosporamidites containing [Ru(acac)(2)(IMPy)](2+) (acac is acetylacetonato; IMPy is 2'-iminomethylpyridyl-2'-deoxyuridine) (3) and [Ru(bpy)(2)(IMPy)](2+) (bpy is 2,2'-bipyridine; IMPy is 2'-iminomethylpyridyl-2' deoxyuridine) (4). Duplexes formed with the metal-containing oligonucleotides exhibit thermal stability comparable to the corresponding unmetalated duplexes (T(m) of modified duplex = 49 degrees C vs T(m) of unmodified duplex = 47 degrees C). Electrochemical (3, E(1/2) = -0.04 V vs NHE; 4, E(1/2) = 1.12 V vs NHE), absorption (3, lambda(max) = 568, 369 nm; 4, lambda(max) = 480 nm), and emission (4, lambda(max) = 720 nm, tau = 55 ns, Phi = 1.2 x 10(-)(4)) data for the ruthenium-modified nucleosides and oligonucleotides indicate that incorporation into an oligonucleotide does not perturb the electronic properties of the ruthenium complex or the DNA significantly. In addition, the absence of any change in the emission properties upon metalated duplex formation suggests that the [Ru(bpy)(2)(IMPy)](2+)[Ru(acac)(2)(IMPy)](2+) pair will provide a valuable probe for DNA-mediated electron-transfer studies. PMID- 12588138 TI - Cationic sigma-phenylplatinum(II) complexes with carboxylic acid functionality: pK(a) determinations and x-ray structures. AB - The preparation and characterization of a novel series of cationic sigma phenylplatinum(II) complexes of the type trans-[Pt(sigma-C(6)H(5))(L)(2)A]OTf (A = picolinic acid, L = PPh(3) (4) and PMePh(2) (7); A = nicotinic acid, L = PPh(3) (5) and PMePh(2) (8); A = isonicotinic acid, L = PPh(3) (6), PMePh(2) (9), and PEt(3) (10)) are described. The pK(a) value for the carboxylic acid functionality in selected complexes was found to follow the order 7 (pK(a) = 5.23 +/- 0.09) > 8 (4.85 +/- 0.10) > 9 (3.51 +/- 0.08) > 6 (3.26 +/- 0.07) approximately 10 (3.21 +/ 0.08) by means of potentiometric titration experiments in 50% (v/v) EtOH/H(2)O solution at 295 K. The X-ray crystal structures of 9 and 10 were also determined. The asymmetric unit of each of 9 and 10 comprises a univalent complex cation, a triflate anion, and a solvent CH(2)Cl(2) molecule of crystallization. Centrosymmetrically related pairs of complex cations in 9 associate via the familiar carboxylic acid dimer motif, whereas with 10, the carboxylic acid dimer motif is absent. Instead, the carboxylic acid residue forms both donor and acceptor interactions to the triflate anion and CH(2)Cl(2) solvent of crystallization, respectively, to afford a 10-membered ring structure. Possible reasons for the observed differences in the solid-state structures of 9 and 10 are presented. PMID- 12588139 TI - Heterobimetallic oxalato-bridged M(II)Re(IV) complexes (M = Mn, Fe, Co, Ni): synthesis, crystal structure, and magnetic properties. AB - Four rhenium(IV)-M(II) bimetallic complexes of formula [ReCl(4)(mu ox)M(dmphen)(2)].CH(3)CN with M = Mn (1), Fe (2), Co (3), and Ni (4) (ox = oxalate anion, dmphen = 2,9-dimethyl-1,10-phenanthroline) have been synthesized and the crystal structures of 1 and 3 determined by single-crystal X-ray diffraction. 1 and 3 are isostructural and crystallize in the monoclinic system, space group P2(1)/c, with a = 16.008(4) A, b = 12.729(2) A, c = 18.909(5) A, beta = 112.70(2) degrees, and Z = 4 for 1 and a = 15.998(4) A, b = 12.665(2) A, c = 18.693(5) A, beta = 112.33(2) degrees, and Z = 4, for 3. The structure of 1 and 3 is made up of neutral [ReCl(4)(mu-ox)M(dmphen)(2)] bimetallic units (M = Mn (1), Co (3)) and acetonitrile molecules of crystallization. M(II) and Re(IV) metal ions exhibit distorted octahedral coordination geometries being bridged by a bis(bidentate) oxalato ligand. The magnetic behavior of 1-4 has been investigated over the temperature range 2.0-300 K. A very weak antiferromagnetic coupling between Re(IV) and Mn(II) occurs in 1 (J = -0.1 cm(-)(1)), whereas a significant ferromagnetic interaction between Re(IV) and M(II) is observed in 2-4 [J = +2.8 (2), +5.2 (3), and +5.9 cm(-)(1) (4)]. PMID- 12588140 TI - Coordination of BF(4)(-) to oxovanadium(V) complexes, evidenced by the redox potential of oxovanadium(IV/V) couples in CH(2)Cl(2). AB - The oxidation of oxovanadium(IV) complexes [LV(IV)O] (L = tetradentate Schiff base ligands such as N,N'-ethylenebis(salicylideneaminate)(2-) (salen) and N,N' 2,2-dimethylpropylenebis(salicylideneaminate)(2-) (salpn)) to [LV(V)O](+), believed to be responsible for the voltammetric response near 0.6 V vs Ag/AgCl in CH(2)Cl(2) in the presence of tetrabutylammonium tetrafluoroborate as a supporting electrolyte, is in fact coupled to a homogeneous process where [LVO](+) coordinates BF(4)(-) to form a neutral complex formulated as [LVOBF(4)]. The formation constants for [VO(salen)BF(4)] and [VO(salpn)BF(4)] are evaluated to be K(salen)(-)(1) = 1.1 x 10(2) M(-)(1) and K(salpn)(-)(1) = 1.4 x 10 M(-)(1), respectively. Crystal structure of [VO(salen)BF(4)] reveals that one of the fluorine atoms in BF(4)(-) is so close to the vanadium(V) atom as to be practically bound in the solid state. PMID- 12588141 TI - Reduction pathway of end-on coordinated dinitrogen. 3. Electronic structure and spectroscopic properties of molybdenum/tungsten hydrazidium complexes. AB - The spectroscopic properties and electronic structure of the hydrazidium complexes [MF(NNH(3))(depe)(2)](BF(4))(2), M = Mo and W, are investigated (depe = 1,2-bis(diethylphosphino)ethane). Vibrational spectroscopic data for both compounds are evaluated with a quantum-chemistry-assisted normal coordinate analysis, giving an N-N force constant of 6.03 mdyn/A and metal-N force constants of 8.01 (Mo-N) and 7.31 mdyn/A (W-N), respectively. On the basis of these results and DFT calculations on a [MoF(NNH(3))(PH(3))(4)](2+) model system, the N-N bond order in these systems is 1 (single sigma bond) and metal-N bonding corresponds to a triple bond. The metal centers are assigned a +IV oxidation state (d(2) configuration) and the NNH(3) ligand is assigned a -1 formal charge which by sigma- and pi-donation to the metal is reduced to +0.48. The two metal-d electrons are located in the nonbonding (n) d(xy)() orbital. This bonding description is supported by the results of optical absorption spectroscopy showing the n --> (metal-ligand)pi transition at 536 nm (not observed in the tungsten compound) and the (metal-ligand)pi --> (metal-ligand)pi transition at 251 nm for the MoNNH(3) and at 237 nm for the WNNH(3) complex. The activation enthalpy for splitting of the N-N bond in these systems to generate NH(4)(+) is estimated to be larger than 40 kcal/mol. Hydrazidium complexes with diphosphine coligands are therefore inert with respect to N-N cleavage and thus represent the ultimate stage of N(2) reduction at six-coordinate d(6) metal centers in the absence of external reductants. PMID- 12588142 TI - Nitrogen ligands on phosphorus(III) Lewis acceptors: A versatile new synthetic approach to unusual N-P structural arrangements. AB - A series of Lewis acid-base adducts of phosphines with amines or pyridines have been prepared and structurally characterized, demonstrating the importance of Lewis acceptor behavior in the chemistry of electron-rich (lone-pair-bearing) centers. The neutral and cationic complexes all contain examples of N-P coordinate bonds that are void of N-P pi-bonding and therefore represent definitive N-P single bonds. The structures are examples of N-P topologies that are rare or have not previously been observed, highlighting the synthetic viability of Lewis acid-base coupling in the still developing field of nitrogen phosphorus chemistry. PMID- 12588143 TI - Syntheses, reactivity, and crystal structures of molybdenum complexes with pyridine-2-thionate (pyS)-containing ligands: crystal structures of [Mo(eta(3) C(3)H(5))(CO)(2)](2)(mu-eta(1),eta(2)-pyS)(2), exo-[Mo(eta(3) C(3)H(5))(CO)(eta(2)-pyS)(eta(2)-dppe)], [Mo(CO)(3)(eta(1)-SC(5)H(4)NH)(eta(2) dppm)], and [Mo(CO)(eta(2)-pyS)(2)(eta(2)-dppm)]. AB - The doubly bridged pyridine-2-thionate (pyS) dimolybdenum complex [Mo(eta(3) C(3)H(5))(CO)(2)](2)(mu-eta(1),eta(2)-pyS)(2) (1) is accessible by the reaction of [Mo(eta(3)-C(3)H(5))(CO)(2)(CH(3)CN)(2)Br] with pySK in methanol at room temperature. Complex 1 reacts with piperidine in acetonitrile to give the complex [Mo(eta(3)-C(3)H(5))(CO)(2)(eta(2)-pyS)(C(5)H(10)NH)] (2). Treatment of 1 with 1,10-phenanthroline (phen) results in the formation of complex [Mo(eta(3) C(3)H(5))(CO)(2)(eta(1)-pyS)(phen)] (3), in which the pyS ligand is coordinated to Mo through the sulfur atom. Four conformational isomers, endo,exo-complexes [Mo(eta(3)-C(3)H(5))(CO)(eta(2)-pyS)(eta(2)-diphos)] (diphos = dppm, 4a-4d; dppe, 5a-5d), are accessible by the reactions of 1 with dppm and dppe in refluxing acetonitrile. Homonuclear shift-correlated 2-D (31)P((1)H)-(31)P((1)H) NMR experiments of the mixtures 4a-4d have been employed to elucidate the four stereoisomers. The reaction of 4 and pySK or [Mo(CO)(3)(eta(1) SC(5)H(4)NH)(eta(2)-dppm)] (6) and O(2) affords allyl-displaced seven-coordinate bis(pyridine-2-thionate) complex [Mo(CO)(eta(2)-pyS)(2)(eta(2)-dppm)] (7). All of the complexes are identified by spectroscopic methods, and complexes 1, 5d, 6, and 7 are determined by single-crystal X-ray diffraction. Complexes 1 and 5d crystallize in the orthorhombic space groups Pbcn and Pbca with Z = 4 and 8, respectively, whereas 6 belongs to the monoclinic space group C2/c with Z = 8 and 7 belongs to the triclinic space group Ponemacr; with Z = 2. The cell dimensions are as follows: for 1, a = 8.3128(1) A, b = 16.1704(2) A, c = 16.6140(2) A; for 5d, a = 17.8309(10) A, b = 17.3324(10) A, c = 20.3716(11) A; for 6, a = 18.618(4) A, b = 16.062(2) A, c = 27.456(6) A, beta = 96.31(3) degrees; for 7, a = 9.1660(2) A, b = 12.0854(3) A, c = 15.9478(4) A, alpha = 78.4811(10) degrees, beta = 80.3894(10) degrees, gamma = 68.7089(11) degrees. PMID- 12588144 TI - Synthesis, crystal structure, and characterization of the first Nb(3) triangular cluster compound bonded to fluorine ligands: association of Nb(3)I(i)F(i)(3)F(a)(8)L(a) units and Nb(IV)L(6) Octahedra with L=O and F. AB - The synthesis and the crystal structure of the first compound containing Nb(3) triangular clusters bonded to fluorine ligands are presented in this work. The structure of Nb(3)IF(7)L(NbL(2))(0.25) with L = O and F, determined by single crystal X-ray diffraction, is based on a Nb(3)I(i)F(i)(3)F(a)(8)L(a) unit and a NbL(6) octahedron (tetragonal, space group I4/m, a = 13.8638(3) A, c = 8.9183(2) A, V = 1714.14(7) A(3), Z = 8). Two crystallographic positions (noted L5 and L6) are randomly occupied by fluorine and oxygen with two different F:O occupancies. These L ligands build an octahedral site for a single niobium atom, located between the units. The four L5 ligands of the NbL(6) octahedron are shared with four Nb(3) cluster units, while the two other L6 ligands are terminal. The Nb(3) cluster is face-capped by one iodine and edge-bridged by three fluorine ligands. Two of the three niobium atoms constituting the cluster are bonded to three additional apical fluorine ligands, while the third one is bonded to two fluorines and one L5 ligand. The Nb(3) cluster is linked to six adjacent ones via all the apical fluorine ligands. The developed formula of the unit is therefore Nb(3)I(i)F(i)(3)F(a)(-)(a)(8/2)L(a) according to the Schafer and Schnering notation. The oxidation state of the single niobium and the random distribution of fluorine and oxygen on the two L sites will be discussed on the basis of structural analysis, the bond valence method, and IR and EPR measurements. The structural results will be compared to those of previously reported niobium compounds containing NbF(6) or Nb(F,O)(6) octahedra. PMID- 12588145 TI - A novel hexanuclear mixed oxidation state Cu(II)(4)Cu(I)(2) cluster complex exhibiting weak ferromagnetic exchange. AB - The complex [(PAH)(4)Cu(II)(4)Cu(I)(2)Br(10)] (1) (PAH = picolinamide hydrazone) containing a Cu(II)(4)Cu(I)(2) hexanuclear cluster, with two well-separated Cu(II)(2) dinuclear centers, results from a redox reaction involving a hydrolytically unstable ligand, salicilyl picolinamide hydrazone, and CuBr(2) in aqueous acetonitrile. The Cu(II) centers are axially bridged via long bromine contacts, leading to ferromagnetic exchange (2J = 4.04 cm(-)(1)). Density functional calculations have been carried out, giving a comparable singlet triplet splitting energy. 1 crystallizes in the triclinic system, space group Ponemacr;, with a = 9.253(3) A, b = 18.159(3) A, c = 7.199(5) A, alpha = 91.31(3) degrees, beta = 107.35(4) degrees, gamma = 104.22(2) degrees, and Z = 2. PMID- 12588146 TI - Anion dependent structures of luminescent silver(i) complexes. AB - The reaction of AgX, where X = trifluoroacetate (CF(3)CO(2)(-), tfa), nitrate (NO(3)(-)), trifluoromethanesulfonate (triflate, CF(3)SO(3)(-), OTf), hexafluorophosphate (PF(6)(-)), or perchlorate (ClO(4)(-)), with 2,2',3' ' tripyridylamine (tpa) yields five novel silver(I) complexes, which have been structurally characterized. The five complexes have the same 1:1 stoichiometry of Ag/tpa but exhibit different modes of coordination, depending upon the counterion present in the compound. Compound 1, [Ag(tpa)(tfa)](n)(), forms a 1D coordination polymer of [Ag(tpa)(tfa)](2) dimer units linked through bridging tfa counterions. Compound 2, [Ag(tpa)(CH(3)CN)(NO(3))](n), forms a zigzag chain 1D coordination polymer exclusively through Ag-N bonds. In compounds 1 and 2, each tpa ligand is bound to two Ag(I) ions via a 2-py and a 3-py group. Compound 3, [Ag(tpa)(OTf)](n), forms a ribbonlike 1D coordination polymer, in which each tpa ligand binds to three different silver centers via all three pyridyl groups, and the counterion remains coordinated to the Ag(I) center. Compounds 4, [Ag(tpa)(CH(3)CN)](n)(PF(6))(n), and 5, [Ag(tpa)(CH(3)CN)](n)() (ClO(4))(n), display ribbonlike structures resembling that of 3, except that the counterions are not coordinated. All complexes are luminescent in acetonitrile solution, with emission maxima in the near-UV region (lambda(max) = 366, 368, 367, 367, and 368 nm for 1-5, respectively). At 77 K, the emission maxima are red-shifted to lambda(max) = 452, 453, 450, 450, and 454 nm for 1-5, respectively. PMID- 12588147 TI - Mixed valence in YBaFe(2)O(5). AB - YBaFe(2)O(5) has been synthesized by heating a nanoscale citrate precursor in a carefully controlled reducing environment. Successful synthesis of a single-phase sample can only be achieved in a narrow window of oxygen partial pressures and temperatures. YBaFe(2)O(5) adopts an oxygen-deficient perovskite-type structure, which contains double layers of corner sharing FeO(5) square pyramids separated by Y(3+) ions. At T(N) congruent with 430 K, tetragonal (P4/mmm) and paramagnetic YBaFe(2)O(5) orders antiferromagnetically (AFM) experiencing a slight orthorhombic distortion (Pmmm). Around this temperature, it can be characterized as a class-III mixed valence (MV) compound, where all iron atoms exist as equivalent MV Fe(2.5+) ions. The magnetic structure is characterized by AFM Fe-O Fe superexchange coupling within the double layers and a ferromagnetic Fe-Fe direct-exchange coupling between neighboring double layers. Upon cooling below approximately 335 K, a premonitory charge ordering (2Fe(2.5+) --> Fe(2.5+delta) + Fe(2.5)(-delta)) into a class-II MV phase takes place. This transition is detected by differential scanning calorimetry, but powder diffraction techniques fail to detect any volume change or a long-range structural order. At approximately 308 K, a complete charge ordering (2Fe(2.5+) --> Fe(2+) + Fe(3+)) into a class-I MV compound takes place. This charge localization triggers a number of changes in the crystal, magnetic, and electronic structure of YBaFe(2)O(5). The magnetic structure rearranges to a G-type AFM structure, where both the Fe-O-Fe superexchange and the Fe-Fe direct-exchange couplings are antiferromagnetic. The crystal structure rearranges (Pmma) to accommodate alternating chains of Fe(2+) and Fe(3+) running along b and an unexpectedly large cooperative Jahn-Teller distortion about the high-spin Fe(2+) ions. This order of charges does not fulfill the Anderson condition, and it rather corresponds to an ordering of doubly occupied Fe(2+) d(xz) orbitals. Comparisons with YBaMn(2)O(5) and YBaCo(2)O(5) are made to highlight the impact of changing the d-electron count. PMID- 12588148 TI - Double-stranded W/Ag/S chain structure generated from [syn-(WO(NCS)(2))(2)(mu S)(2)] building blocks. AB - The dimeric W(V) complex [Et(4)N](4)[syn-(O=W(NCS)(3))(2)(mu-S)(2)], 1, prepared from [Et(4)N](2)[WS(4)], SCN(-), and Cd(2+), shows interesting reactivity patterns in that the thiocyanate trans to the oxo group can in part be replaced, initiated by Mn(2+), by dimethylformamide (DMF) to form [Et(4)N](2.5)[(O=W(NCS)(2.25)(DMF)(1.25))(2)(mu-S)(2)], 2. With Ag(+), 1 undergoes partial replacement of SCN(-) by DMF and coordinates to the silver ions to generate ([Et(4)N](2.5)[(W(2)O(2)(NCS)(2)(mu-S)(2))(mu NCS)(2)(DMF)(Ag(0.5)(SCN))])(n), 3. Compound 3 constitutes a polymeric double stranded chain, with normal bonding interactions [via W-(mu-NCS)-Ag] between the two strands, and moderate intrastrand [W-(mu-NCS).Ag] bonding. The crystal and molecular structures of the three compounds are described. PMID- 12588149 TI - Heteropolymolybdates of phosphate, phosphonate, and phosphite functionalized by glycine. AB - The novel, functionalized heteropolymolybdates [RPMo(6)O(21)(O(2)CCH(2)NH(3))(3)](2)(-) (R = OH, CH(3), C(2)H(5), H) have been synthesized and characterized by IR, (31)P NMR spectroscopy, and elemental analysis. Single-crystal X-ray analysis was carried out on K(2)[HOPMo(6)O(21)(O(2)CCH(2)NH(3))(3)].8.5H(2)O, which crystallizes in the orthorhombic system, space group Pnma, with a = 14.118(2) A, b = 20.660(3) A, c = 12.191(2) A, and Z = 4; K(2)[H(3)CPMo(6)O(21)(O(2)CCH(2)NH(3))(3)].8.5H(2)O, which crystallizes in the orthorhombic system, space group Pnma, with a = 14.1643(6) A, b = 20.8658(8) A, c = 12.2235(5) A, and Z = 4; and K(2)[HPMo(6)O(21)(O(2)CCH(2)NH(3))(3)].8H(2)O, which crystallizes in the orthorhombic system, space group Pnma, with a = 14.092(3) A, b = 20.696(2) A, c = 12.199(4) A, and Z = 4. We also report on the synthesis and characterization of the isostructural derivative K(2)[H(5)C(2)PMo(6)O(21)(O(2)CCH(2)NH(3))(3)]. The four title polyanions consist of an RP (R = OH, CH(3), C(2)H(5), H) hetero group surrounded by a ring of six MoO(6) octahedra sharing edges and corners alternatingly. Three glycine molecules are each bound to two edge-sharing Mo centers via their carboxylate functionality on the same side of the ring. The central phosphorus atom is located slightly above the plane of the six molybdenums, and its terminal R group is on the same side of the ring as the glycines. NMR studies show that the solid state structures of the title compounds are preserved in solution. PMID- 12588150 TI - Tris(tert-butoxy)siloxy derivatives of boron, including the boronous acid HOB[OSi(O(t)Bu)(3)](2) and the metal (siloxy)boryloxide complex Cp(2)Zr(Me)OB[OSi(O(t)Bu)(3)](2): a remarkable crystal structure with 18 independent molecules in its asymmetric unit. AB - Silanolysis of B(O(t)Bu)(3) with 2 and 3 equiv of HOSi(O(t)Bu)(3) led to the formation of (t)BuOB[OSi(O(t)Bu)(3)](2) (1) and B[OSi(O(t)Bu)(3)](3) (2), respectively. Compounds 1 and 2 are efficient single-source molecular precursors to B/Si/O materials via thermolytic routes in nonpolar media, as demonstrated by the generation of BO(1.5).2SiO(2) (BOSi2(xg)) and BO(1.5).3SiO(2) (BOSi3(xg)) xerogels, respectively. Use of a block copolymer template provided B/Si/O materials (BOSi2(epe) and BOSi3(epe)) with a broad distribution of mesopores (by N(2) porosimetry) and smaller, more uniform particle sizes (by TEM) as compared to the nontemplated materials. Hydrolyses of 1 and 2 with excess H(2)O resulted in formation of the expected amounts of (t)BuOH and HOSi(O(t)Bu)(3); however, reaction of 1 with 1 equiv of H(2)O led to isolation of the new boronous acid HOB[OSi(O(t)Bu)(3)](2) (3). This ligand precursor is well suited for the synthesis of new metal (siloxy)boryloxide complexes via proton-transfer reactions involving the BOH group. The reaction of 3 with Cp(2)ZrMe(2) resulted in formation of Cp(2)Zr(Me)OB[OSi(O(t)Bu)(3)](2) (4) in high yield. This rare example of a transition metal boryloxide complex crystallizes in the triclinic space group Ponemacr; and exhibits a crystal structure with an unprecedented number of independent molecules in its asymmetric unit (i.e., Z' = 18 and Z = 36). This unusual crystal structure presented an opportunity to perform statistical analyses of the metric parameters for the 18 crystallographically independent molecules. Complex 4 readily converts to Cp(2)Zr[OSi(O(t)Bu)(3)](2) (5) upon thermolysis or upon dissolution in Et(2)O at room temperature. PMID- 12588151 TI - Nickel(II) macrocyclic complexes with long alkyl pendant chain: synthesis, x-ray structure, and anion exchange property in the solid state. AB - A nickel(II) pentaaza macrocyclic complex containing a 1-hexadecyl pendant chain, [Ni(C(25)H(55)N(5))](ClO(4))(2).H(2)O (1), was synthesized by a one-pot metal template condensation reaction. Crystal data for 1: triclinic, Ponemacr;, a = 8.333(4) A, b = 8.356(3) A, c = 28.374(9) A, alpha = 81.865(19) degrees, beta = 86.242(18) degrees, gamma = 63.871(17) degrees, Z = 2. Solid 1 forms hydrophobic layers that are constructed by the long alkyl chains of the macrocycles. Solid 1 exchanges ClO(4)(-) with NCS(-), PF(6)(-), C(2)O(4)(2-), NO(3)(-), and CF(3)SO(3)(-) that are dissolved in water. From the reaction of [Ni(C(25)H(55)N(5))Cl(2)] with Et(3)NH(TCNQ)(2) in EtOH/DMF/acetone solution, [Ni(C(25)H(55)N(5))(TCNQ)(2)](TCNQ).(CH(3)COCH(3)) (2) (TCNQ = 7,7,8,8-tetracyano p-quinodimethane) was prepared. Crystal data for 2: triclinic, Ponemacr;, a = 8.459(0) A, b = 13.945 (1) A, c = 26.833(2) A, alpha = 88.744(2) degrees, beta = 84.536(2) degrees, gamma = 80.089(4) degrees, Z = 2. In 2, TCNQ anions coordinate nickel(II) at the axial sites, which form pi-stacked TCNQ(-) dimers to give rise to 1-D chains. The neutral TCNQ molecules are included between the dimerized TCNQ(-) species, which construct a pi-stacked group of six TCNQ units as blocked by the long alkyl chains. Compound 2 is an electric insulator. It shows a weak signal in the EPR spectrum. The magnetic susceptibility data of 2 measured at 5 300 K exhibit a simple paramagnetism at low temperatures (<100 K) but an increase in the magnetic moment at higher temperatures due to the contribution of a thermally accessible triplet state for the antiferromagnetically coupled [TCNQ](2)(2-). PMID- 12588152 TI - Polyoxomolybdates functionalized with phosphonocarboxylates. AB - The novel, functionalized heteropolymolybdate [(O(2)CCH(2)PO(3))(2)Mo(5)O(15)](6 ) (1) has been synthesized and characterized by IR and (31)P NMR spectroscopy and elemental analysis. Single-crystal X-ray analysis was carried out on Rb(4)KNa[(O(2)CCH(2)PO(3))(2)Mo(5)O(15)].H(2)O, which crystallizes in the monoclinic system, space group P2(1)/n, with a = 10.146(2) A, b = 13.704(3) A, c = 20.577(4) A, beta = 94.88(3) degrees, and Z = 4. The title polyanion consists of a ring of five MoO(6) octahedra with four edge junctions and one corner junction. This nonplanar arrangement is stabilized by two phosphonocarboxylate groups that are bound via their phosphonate functionalities on opposite sides of the ring. As a result the two dangling arms with their terminal carboxylate groups protrude away from the molybdenum-oxo framework in diametrically opposed directions. The solid-state structure of 1 is preserved in solution on the basis of NMR. We also report on the synthesis and characterization of the isostructural derivative [(O(2)CC(2)H(4)PO(3))(2)Mo(5)O(15)](6-) (2). PMID- 12588153 TI - Multi-iron tungstodiarsenates. Synthesis, characterization, and electrocatalytic studies of alphabetabetaalpha (Fe(III)OH(2))(2)Fe(III)(2)(As(2)W(15)O(56))(2)(12)(-). AB - Reaction of the trivacant lacunary complex, alpha-Na(12)[As(2)W(15)O(56)], with an aqueous solution of Fe(NO(3))(3).9H(2)O yields the sandwich-type polyoxometalate, alphabetabetaalpha Na(12)(Fe(III)OH(2))(2)Fe(III)(2)(As(2)W(15)O(56))(2) (Na1). The structure of this complex, determined by single-crystal X-ray crystallography (a = 13.434(1) A, b = 13.763(1) A, c = 22.999(2) A, alpha = 90.246(2) degrees, beta = 102.887(2) degrees, gamma = 116.972(1) degrees, triclinic, Ponemacr;, R1 = 5.5%, based on 25342 independent reflections), consists of an Fe(III)(4) unit sandwiched between two trivacant alpha-As(2)W(15)O(56)(12)(-) moieties. UV-vis, infrared, cyclic voltammetry, and elemental analysis data are all consistent with the structure determined from X-ray analysis. Magnetization studies confirm that the four Fe(III) centers are antiferromagnetically coupled. A cyclic voltammogram of Na1 reveals that a three-wave W(VI) system replaces the two-wave W(VI) system found in the precursor alpha-As(2)W(15)O(56)(12)(-) complex. The observed modifications in the CV patterns of Na1 and alpha-As(2)W(15)O(56)(12)(-) are most likely due to subsequent changes in the acid-base properties of two reduced POMs that occur as a result of Fe(III) incorporation. Na1 is shown to be more efficient than the monosubstituted complex alpha(2)-As(2)(Fe(III)OH(2))W(17)O(61)(7)(-) in the electrocatalytic reduction of dioxygen. This is attributed to cooperativity effects among the adjacent Fe(III) centers in Na1. PMID- 12588154 TI - An organically templated cobalt-vanadium oxide with beta cage units: hydrothermal synthesis and x-ray structural characterization of (C(2)H(10)N(2))[Co(2)(C(2)O(4))V(4)O(12)]. AB - A novel organically templated cobalt-vanadium oxalate, (C(2)H(10)N(2))[Co(2)(C(2)O(4))V(4)O(12)], was synthesized under mild hydrothermal conditions and characterized by single-crystal/powder X-ray diffraction, infrared spectroscopy, and thermogravimetric analysis. The compound crystallizes in an orthorhombic system with space group Cmcm and cell parameters a = 11.527(2) A, b = 9.9476(18) A, c = 14.780(3) A. The compound possesses 3 dimensional topologies with sodalite analogue structure and is constructed by C(2)O(4)-incorporated beta cage units. On the basis of the results of TG/DTA analyses, the structure is thermally stable up to approximately 573 K. PMID- 12588155 TI - The solid state structure of [b(10)h(11)](-) and its dynamic NMR spectra in solution. AB - The structure of [PPh(3)(benzyl)][B(10)H(11)] was determined at -123 degrees C and 24 degrees C by single-crystal X-ray analyses. The B(10) core of [B(10)H(11)](-) is similar in shape to that of [B(10)H(10)](2)(-). The 11th H atom asymmetrically caps a polar face of the cluster and shows no tendency for disorder in the solid state. Variable temperature multinuclear NMR studies shed light on the dynamic nature of [B(10)H(11)](-) in solution. In addition to the fluxionality of the cluster H atoms, the boron cage is fluxional at moderate temperatures, in contrast to [B(10)H(10)](2)(-). Multiple exchange processes are believed to take place as a function of temperature. Results of ab initio calculations are presented. Crystal data: [PPh(3)(benzyl)][B(10)H(11)] at -123 degrees C, P2(1)/c, a = 9.988(2) A, b = 18.860(2) A, c = 15.072(2) A, beta = 107.916(8) degrees, V = 2701.5(7) A(3), Z = 4; [PPh(3)(benzyl)][B(10)H(11)] at 24 degrees C, P2(1)/c, a = 10.067(5) A, b = 19.009(9) A, c = 15.247(7) A, beta = 107.952(9) degrees, V = 2775(2) A(3), Z = 4. PMID- 12588156 TI - Magnetic superstructures of cupric oxide CuO as ordered arrangements of one dimensional antiferromagnetic chains. AB - In cupric oxide CuO, each Cu(2+) ion has 12 nearest-neighbor Cu(2+) ions grouped into six pairs related by inversion symmetry. The relative strengths of the Cu-O Cu superexchange interactions in cupric oxide CuO were estimated by spin dimer analysis to confirm that the strongest superexchange interactions form one dimensional antiferromagnetic chains along the [101] direction, and the remaining interactions are weak. We analyzed ordered arrangements of these one-dimensional antiferromagnetic chains to examine why the antiferromagnetic phase transition of CuO below 212.5 K adopts a (2a, b, 2c) superstructure. The local spin arrangement around each Cu(2+) ion is more balanced in the ordered spin structures leading to a (2a, b, 2c) supercell than in any other ordered spin structures. PMID- 12588157 TI - Structural modulation of Cu(I) and Cu(II) complexes of sterically hindered tripyridine ligands by the bridgehead alkyl groups. AB - Structures of Cu(I) and Cu(II) complexes of sterically hindered tripyridine ligands RL = tris(6-methyl-2-pyridyl)methane (HL), 1,1,1-tris(6-methyl-2 pyridyl)ethane (MeL), and 1,1,1-tris(6-methyl-2-pyridyl)propane (EtL), [Cu(RL)(MeCN)]PF(6) (1-3), [Cu(RL)(SO(4))] (4-6), and [Cu(RL)(NO(3))(2)] (7-9), have been explored in the solid state and in solution to gain some insights into modulation of the copper coordination structures by bridgehead alkyl groups (CH, CMe, and CEt). The crystal structures of 1-9 show that RL binds a copper ion in a tridentate facial-capping mode, except for 3, where EtL chelates in a bidentate mode with two pyridyl nitrogen atoms. To avoid the steric repulsion between the bridgehead alkyl group and the 3-H(py) atoms, the pyridine rings in Cu(I) and Cu(II) complexes of MeL and EtL shift toward the Cu side as compared to those in Cu(I) and Cu(II) complexes of HL, leading to the significant differences in the nonbonding interatomic distances, H.H (between the 3-H(py) atoms), N.N (between the N(py) atoms), and C.C (between the 6-Me carbon atoms), the Cu-N(py), Cu N(MeCN), and Cu-O bond distances, and the tilt of the pyridine rings. The copper coordination geometries in 4-6, where a SO(4) ligand chelates in a bidentate mode, are varied from a square pyramid of 4 to distorted trigonal bipyramids of 5 and 6. Such structural differences are not observed for 7-9, where two NO(3) ligands coordinate in a monodentate mode. The structures of 1-9 in solution are investigated by means of the electronic, (1)H NMR, and ESR spectroscopy. The (1)H NMR spectra show that the structures of 1-3 in the solid state are kept in solution with rapid coordination exchange of the pyridine rings. The electronic and the ESR spectra reveal the structural changes of 5 and 6 in solution. The bridgehead alkyl groups and 6-Me groups in the sterically hindered tripyridine ligand play important roles in modulating the copper coordination structures. PMID- 12588158 TI - New insights into an old reaction. High-resolution x-ray powder diffraction of Wiberg's aminoalane intermediate. AB - In accordance with the procedure described by E. Wiberg, Me(3)Al-NH(3) was heated as a bulk material in inert atmosphere to give a colorless liquid which slowly loses methane. Close to the end of this elimination reaction, the melt crystallized to give a microcrystalline powder of (Me(2)AlNH(2))(x)(). The structure of this intermediate has been solved by the method of high-resolution X ray powder diffraction. The compound crystallizes in the monoclinic space group C2/c with the cell parameters of a = 15.0047(6) A, b = 8.7500(2) A, c = 24.4702(8) A, and beta = 107.290(2) degrees, with eight trimers (Me(2)AlNH(2))(3) per unit cell. These trimers crystallize in a boat conformation in contrast to the known trimers of the same composition where a twist-boat conformation had been found by single crystal determination. Different conformers of (Me(2)AlNH(2))(3) have been investigated by theoretical methods (HF/6-31G(d), B3LYP/6-31G(d), B3LYP/6-311G(d,p), MP2(fc)/6-31G(d), and MP2(fc)/6-311G(d,p)). The twist-boat and the chair conformer correspond to minima at the potential energy surface, whereas the boat conformer corresponds to a first-order transition state (relative energies of 0.45-2.56 kJ/mol (boat) and 6.66-11.91 kJ/mol (chair)). Relaxed scans of the potential energy surface at the HF/6-31G(d) and B3LYP/6-31G(d) levels have shown that the boat conformer (C(s)() symmetry) connects two enantiomers of the twist-boat form (C(2) symmetry). PMID- 12588159 TI - Iron-arylimide clusters [Fe(m)()(NAr)(n)Cl(4)](2)(-) (m, n = 2, 2; 3, 4; 4, 4) from a ferric amide precursor: synthesis, characterization, and comparison to Fe S chemistry. AB - Tetrahedral FeCl[N(SiMe(3))(2)](2)(THF) (2), prepared from FeCl(3) and 2 equiv of Na[N(SiMe(3))(2)] in THF, is a useful ferric starting material for the synthesis of weak-field iron-imide (Fe-NR) clusters. Protonolysis of 2 with aniline yields azobenzene and [Fe(2)(mu-Cl)(3)(THF)(6)](2)[Fe(3)(mu-NPh)(4)Cl(4)] (3), a salt composed of two diferrous monocations and a trinuclear dianion with a formal 2 Fe(III)/1 Fe(IV) oxidation state. Treatment of 2 with LiCl, which gives the adduct [FeCl(2)(N(SiMe(3))(2))(2)](-) (isolated as the [Li(TMEDA)(2)](+) salt), suppresses arylamine oxidation/iron reduction chemistry during protonolysis. Thus, under appropriate conditions, the reaction of 1:1 2/LiCl with arylamine provides a practical route to the following Fe-NR clusters: [Li(2)(THF)(7)][Fe(3)(mu-NPh)(4)Cl(4)] (5a), which contains the same Fe-NR cluster found in 3; [Li(THF)(4)](2)[Fe(3)(mu-N-p-Tol)(4)Cl(4)] (5b); [Li(DME)(3)](2)[Fe(2)(mu-NPh)(2)Cl(4)] (6a); [Li(2)(THF)(7)][Fe(2)(mu NMes)(2)Cl(4)] (6c). [Li(DME)(3)](2)[Fe(4)(mu(3)-NPh)(4)Cl(4)] (7), a trace product in the synthesis of 5a and 6a, forms readily as the sole Fe-NR complex upon reduction of these lower nuclearity clusters. Products were characterized by X-ray crystallographic analysis, by electronic absorption, (1)H NMR, and Mossbauer spectroscopies, and by cyclic voltammetry. The structures of the Fe-NR complexes derive from tetrahedral iron centers, edge-fused by imide bridges into linear arrays (5a,b; 6a,c) or the condensed heterocubane geometry (7), and are homologous to fundamental iron-sulfur (Fe-S) cluster motifs. The analogy to Fe-S chemistry also encompasses parallels between Fe-mediated redox transformations of nitrogen and sulfur ligands and reductive core conversions of linear dinuclear and trinuclear clusters to heterocubane species and is reinforced by other recent examples of iron- and cobalt-imide cluster chemistry. The correspondence of nitrogen and sulfur chemistry at iron is intriguing in the context of speculative Fe-mediated mechanisms for biological nitrogen fixation. PMID- 12588160 TI - Kinetics and mechanisms of the oxidation of phenols by a trans-dioxoruthenium(VI) complex. AB - The kinetics of the oxidation of phenols by trans-[Ru(VI)(L)(O)(2)](2+) (L = 1,12 dimethyl-3,4:9,10-dibenzo-1,12-diaza-5,8-dioxacyclopentadecane) have been studied in aqueous acidic solutions and in acetonitrile. In H(2)O the oxidation of phenol produces the unstable 4,4'- biphenoquinone, as evidenced by a rapid increase and then a slow decrease in absorbance at 398 nm. The first step is first-order in both Ru(VI) and phenol, and rate constants are dependent on [H(+)] according to the relationship k(f) = k(x) + (k(y)K(a)/[H(+)]), where k(x) and k(y) are the rate constants for the oxidation of PhOH and PhO(-), respectively. At 298 K and I = 0.1 M, k(x) = 12.5 M(-1) s(-1) and k(y) = 8.0 x 10(8) M(-1) s(-1). At I = 0.1 M and pH = 2.98, the kinetic isotope effects are k(H(2)O)/k(D(2)O) = 4.8 and 0.74 for k(x) and k(y), respectively, and k(f)(C(6)H(5)OH)/k(f)(C(6)D(5)OH) = 1.1. It is proposed that the k(x) step occurs by a hydrogen atom abstraction mechanism, while the k(y) step occurs by an electron-transfer mechanism. In both steps the phenoxy radical is produced, which then undergoes two rapid concurrent reactions. The first is a further three-electron oxidation by Ru(VI) and Ru(V) to give p benzoquinone and other organic products. The second is a coupling and oxidation process to give 4,4'-biphenoquinone, followed by the decay step, k(s). A similar mechanism is proposed for reactions in CH(3)CN. A plot of log k(x) vs O-H bond dissociation enthalpies (BDE) of the phenols separates those phenols with bulky tert-butyl substituents in the ortho positions from those with no 2,6-di-tert butyl groups into two separate lines. This arises because there is steric crowding of the hydroxylic groups in 2,6-di-tert-butyl phenols, which react more slowly than phenols of similar O-H BDE but no 2,6-tert-butyl groups. This is as expected if hydrogen atom abstraction but not electron transfer is occurring. PMID- 12588161 TI - Solution chemistry of uranyl ion with iminodiacetate and oxydiacetate: A combined NMR/EXAFS and potentiometry/calorimetry study. AB - The solution chemistry of uranyl ion with iminodiacetate (IDA) and oxydiacetate (ODA) was investigated using NMR and EXAFS spectroscopies, potentiometry, and calorimetry. From the NMR and EXAFS data and depending on stoichiometry and pH, three types of metal:ligand complex were identified in solution in the pH range 3 7: 1:1 and 1:2 monomers; a 2:2 dimer. From NMR and EXAFS data for the IDA system and previous studies, we propose the three complex types are [UO(2)(IDA)(H(2)O)(2)], [UO(2)(IDA)(2)](2)(-), and [(UO(2))(2)(IDA)(2)(mu OH)(2)](2)(-). From EXAFS spectroscopy, similar 1:1, 2:2, and 1:2 complexes are found for the ODA system, although (13)C NMR spectroscopy was not a useful probe in this system. For the 1:1 and 1:2 complexes in solution, EXAFS spectroscopy is ambiguous because the data can be fitted with either a long U-N/O(ether) value (ca. 2.9 A) suggesting 1,7-coordination of the ligand or a U-C interaction at a similar distance, consistent with terminal bidentate coordination. However, the NMR data of the IDA system suggest that 1,7-coordination is the more likely. The stability constants of the three complexes were determined by potentiometric titrations; the log beta values are 9.90 +/-, 16.42 +/-, and 10.80 +/- for the 1:1, 1:2, and 2:2 uranyl-IDA complexes, respectively, and 5.77 +/-, 7.84 +/-, and 4.29 +/- for the 1:1, 1:2, and 2:2 uranyl-ODA complexes, respectively. The thermodynamic constants for the complexes were calculated from calorimetric titrations; the enthalpy changes (kJ mol(-)(1)) and entropy changes (J K(-)(1) mol(-)(1)) of complexation for the 1:1, 1:2, and 2:2 complexes respectively are the following. IDA: 12 +/- 2, 230 +/- 8; 8 +/- 2, 151 +/- 9; -33 +/- 3, -283 +/- 11. ODA: 26 +/- 2, 198 +/- 12; 20 +/- 2, 106 +/- 8; -24 +/- 2; -219 +/- 8. PMID- 12588162 TI - Complexation of the triply-bonded dirhenium(II) complex Re(2)Cl(4)(mu-dppm)(2) (dppm = Ph(2)PCH(2)PPh(2)) by up to three acetylene molecules. AB - The triply bonded dirhenium(II) synthons Re(2)X(4)(mu-dppm)(2) (X = Cl, Br; dppm = Ph(2)PCH(2)PPh(2)) react with acetylene at room temperature in CH(2)Cl(2) and acetone to afford the bis(acetylene) complexes Re(2)X(4)(mu dppm)(2)(mu:eta(2),eta(2)-HCCH)(eta(2)-HCCH) (X = Cl (3), Br(4)). Compound 3 has been derivatized by reaction with RNC ligands in the presence of TlPF(6) to give unsymmetrical complexes of the type [Re(2)Cl(3)(mu-dppm)(2)(mu:eta(2),eta(2) HCCH)(eta(2)-HCCH)(CNR)]PF(6) (R = Xyl (5), Mes (6), t-Bu (7)), in which the RCN ligand has displaced the chloride ligand cis to the eta(2)-HCCH ligand. The reaction of 3 with an additional 1 equiv of acetylene in the presence of TlPF(6) gives the symmetrical all-cis isomer of [Re(2)Cl(3)(mu-dppm)(2)(mu:eta(2),eta(2) HCCH)(eta(2)-HCCH)(2)]PF(6) (8). The two terminal eta(2)-HCCH ligands in 8 are very labile and can be displaced by CO and XylNC to give the complexes [Re(2)Cl(3)(mu-dppm)(2)(mu:eta(2),eta(2)-HCCH)(L)(2)]Y (L = CO when Y = PF(6) (9); L = CO when Y = (PF(6))(0.5)/(H(2)PO(4))(0.5) (10); L = XylNC when Y = PF(6) (11)). These substitution reactions proceed with retention of the all-cis stereochemistry. Single-crystal X-ray structure determinations have been carried out on complexes 3, 5, 8, 10, and 11. In no instance have we found that the acetylene ligands undergo reductive coupling reactions. PMID- 12588163 TI - Realizing green phosphorescent light-emitting materials from rhenium(i) pyrazolato diimine complexes. AB - Two neutral pyrazolato diimine rhenium(I) carbonyl complexes with formula [Re(CO)(3)(N-N)(btpz)] where N-N = 2,2'-bipyridine (1) and 1,10-phenanathroline (2), and btpz = 3,5-bis(trifluoromethyl) pyrazolate, were synthesized and characterized by elemental analysis, routine spectroscopic methods, and single crystal X-ray diffraction study. Ground and excited state properties of these complexes were investigated by steady-state and time-resolved spectroscopies. Complexes 1 and 2 show photoluminescent emission in both solution and solid-state at room temperature, arising from metal to ligand charge-transfer (MLCT) transition with strong overlapping of intraligand pi --> pi transitions. The long lived excited state lifetimes of complexes 1 and 2, which are on the order of microseconds, indicate the presence of phosphorescent emission. As these complexes hold the potential to serve as phosphors for organic light-emitting diodes (OLEDs), their electroluminescent performances were evaluated by employing them as dopants of various electron transport layer (ETL) or hole transport layer (HTL) hosts. For complex 1, a green electrophosphorescence emission centered at lambda(max) = 530 nm was observed at low turn-on voltage ( approximately 6 V) with luminous power efficiency of 0.72 lm/W, external quantum efficiency of 0.82%, and luminance of 2300 cd/m(2) at a current density of 100 mA/cm(2). PMID- 12588164 TI - 51V solid-state magic angle spinning NMR spectroscopy and DFT studies of oxovanadium(V) complexes mimicking the active site of vanadium haloperoxidases. AB - A series of 11 oxovanadium(V) complexes mimicking the active site of vanadium haloperoxidases have been investigated by (51)V magic angle spinning NMR spectroscopy and density functional theory (DFT). The MAS spectra are dominated by the anisotropic quadrupolar and chemical shielding interactions; for these compounds, C(Q) ranges from 3 to 8 MHz, and delta(sigma) is in the range 340-730 ppm. The quadrupolar coupling and chemical shielding tensors as well as their relative orientations have been determined by numerical simulations of the spectra. The spectroscopic NMR observables appear to be very sensitive to the details of the electronic and geometric environment of the vanadium center in these complexes. For the four crystallographically characterized compounds from the series, the quadrupolar and chemical shielding anisotropies were computed at the DFT level using two different basis sets, and the calculated tensors were in general agreement with the experimental solid-state NMR data. A combination of (51)V solid-state NMR and computational methods is thus beneficial for investigation of the electrostatic and geometric environment in diamagnetic vanadium systems with moderate quadrupolar anisotropies. PMID- 12588165 TI - Inhibition of transcription in vitro by anticancer active dirhodium(II) complexes. AB - The DNA binding and inhibition of transcription in vitro by neutral Rh(2)(mu O(2)CCH(3))(4) and cationic cis-[Rh(2)(mu-O(2)CCH(3))(2)(phen)(2)](2+) complexes were investigated. The binding constants of the two complexes to calf-thymus DNA were estimated from absorption titrations to be 4.6 x 10(2) M(-)(1) and 1.7 x 10(4) M(-)(1) for Rh(2)(mu-O(2)CCH(3))(4) and cis-[Rh(2)(mu O(2)CCH(3))(2)(phen)(2)](2+), respectively. The shift to higher energies of the low-energy absorption of the complexes upon addition of DNA is consistent with axial binding of the complexes to duplex DNA. The relative concentrations, [complex]/[DNA], of Rh(2)(mu-O(2)CCH(3))(4) and cis-[Rh(2)(mu O(2)CCH(3))(2)(phen)(2)](2+) at which 50% of the transcription is inhibited (R(inh)(50)), are 0.0031 and 0.0011, respectively. These concentrations are significantly lower than that required for activated cisplatin, cis [Pt(NH(3))(2)(H(2)O)(2)](2+), with R(inh)(50) = 0.0085 under similar experimental conditions. Upon incubation of cis-[Pt(NH(3))(2)(H(2)O)(2)](2+) with the template DNA prior to the addition of the enzyme and nucleobases necessary for the transcription reaction for 30 min at 37 degrees C, significantly lower concentrations of the complex were required to attain 50% inhibition. In contrast, similar incubation of the DNA with the dirhodium complexes did not result in better transcription inhibition. Experiments designed to elucidate the mechanism of the observed inhibition indicate that, unlike cis [Pt(NH(3))(2)(H(2)O)(2)](2+), Rh(2)(mu-O(2)CCH(3))(4) and cis-[Rh(2)(mu O(2)CCH(3))(2)(phen)(2)](2+) appear to interact directly with the enzyme T7-RNA polymerase as their mode of inhibition. PMID- 12588166 TI - Insertion of carbon fragments into P(III)-N bonds in aminophosphines and aminobis(phosphines): synthesis, reactivity, and coordination chemistry of resulting phosphine oxide derivatives. Crystal and molecular structures of (Ph(2)P(O)CH(2))(2)NR (R = Me, (n)Pr, (n)Bu), Ph(2)P(O)CH(OH)(n)()Pr, and cis [MoO(2)Cl(2)((Ph(2)P(O)CH(2))(2)NEt-kappaO,kappaO)]. AB - Reactions of N-aryl and N-alicyclic derivatives of aminophosphines with paraformaldehyde lead to methylene insertion into P-N bond followed by oxidation of phosphorus from the P(III) to P(V) state. When N-alkyl derivatives are reacted with paraformaldehyde, dimerization takes place to afford bis(phosphine oxide)s of the type Ph(2)P(O)CH(2)N(R)CH(2)P(O)Ph(2) (R = Me, (n)Pr, (n)Bu). Aminobis(phosphines) also undergo methylene insertion when treated with paraformaldehyde to give bis(phosphine oxides) Ph(2)P(O)CH(2)N(R)CH(2)P(O)Ph(2) (R = Me, Et, (n)()Pr, (i)()Pr, (n)Bu) in good yield. The reaction of aminophosphines with aromatic aldehydes ArCHO leads to insertion of "ArCH" into the P-N bond to give Ph(2)P(O)CH(R)N(H)Ph (R = C(6)H(5), furfuryl, o-C(6)H(4)OH), but with aliphatic aldehydes such as butanal, P-N bond cleavage takes place to afford alpha-hydroxy phosphine oxide. The reaction of aminobis(phosphines) with both aromatic and aliphatic aldehydes leads to the formation of alpha-hydroxy phosphine oxides through P-N bond cleavage whereas the reaction with furfural leads to the P-N bond insertion. The structure of the alpha-hydroxy derivative Ph(2)P(O)CR(H)(OH)(n)()Pr shows intermolecular hydrogen bonding between OH and P=O oxygen. The phosphine oxide derivatives act as bidentate ligands and form chelate complexes with Co(II), Mo(VI), Th(IV), and U(VI) derivatives. The crystal structure of the molybdenum complex, cis-[MoO(2)Cl(2)((OPPh(2)CH(2))(2)NEt kappaO,kappaO)], shows the distorted octahedral geometry around Mo with two oxo groups cis to each other. PMID- 12588167 TI - Platinum and palladium imido and oxo complexes with small natural bite angle diphosphine ligands. AB - Treatment of L(2)MCl(2) (M = Pt, Pd; L(2) = Ph(2)PCMe(2)PPh(2) (dppip), Ph(2)PNMePPh(2) (dppma)) with AgX (X = OTf, BF(4), NO(3)) in wet CH(2)Cl(2) yields the dinuclear dihydroxo complexes [L(2)M(mu-OH)](2)(X)(2), the mononuclear aqua complexes [L(2)M(OH(2))(2)](X)(2), the mononuclear anion complexes L(2)MX(2), or mixtures of complexes. Addition of aromatic amines to these complexes or mixtures gives the dinuclear diamido complexes [L(2)Pt(mu NHAr)](2)(BF(4))(2), the mononuclear amine complexes [L(2)M(NH(2)Ar)(2)](X)(2), or the dinuclear amido-hydroxo complex [Pt(2)(mu-OH)(mu NHAr)(dppip)(2)](BF(4))(2). Deprotonation of the Pd and Pt amine or diamido complexes with M'N(SiMe(3))(2) (M' = Li, Na, K) gives the diimido complexes [L(2)M(mu-NAr)](2) associated with M' salts. Structural studies of the Li derivatives indicate association through coordination of the imido nitrogen atoms to Li(+). Deprotonation of the amido-hydroxo complex gives the imido-oxo complex [Pt(2)(mu-O)(mu-NAr)(dppip)(2)].LiBF(4).LiN(SiMe(3))(2), and deprotonation of the dppip Pt hydroxo complex gives the dioxo complex [Pt(mu O)(dppip)](2).LiN(SiMe(3))(2).2LiBF(4). PMID- 12588168 TI - Chalcogenide-halides of niobium (V). 1. Gas-phase structures of NbOBr(3), NbSBr(3), and NbSCl(3). 2. Matrix infrared spectra and vibrational force fields of NbOBr(3), NbSBr(3), NbSCl(3), and NbOCl(3). AB - The molecular structures of NbOBr(3), NbSCl(3), and NbSBr(3) have been determined by gas-phase electron diffraction (GED) at nozzle-tip temperatures of 250 degrees C, taking into account the possible presence of NbOCl(3) as a contaminant in the NbSCl(3) sample and NbOBr(3) in the NbSBr(3) sample. The experimental data are consistent with trigonal-pyramidal molecules having C(3)(v)() symmetry. Infrared spectra of molecules trapped in argon or nitrogen matrices were recorded and exhibit the characteristic fundamental stretching modes for C(3)(v)() species. Well resolved isotopic fine structure ((35)Cl and (37)Cl) was observed for NbSCl(3), and for NbOCl(3) which occurred as an impurity in the NbSCl(3) spectra. Quantum mechanical calculations of the structures and vibrational frequencies of the four YNbX(3) molecules (Y = O, S; X = Cl, Br) were carried out at several levels of theory, most importantly B3LYP DFT with either the Stuttgart RSC ECP or Hay-Wadt (n + 1) ECP VDZ basis set for Nb and the 6-311G basis set for the nonmetal atoms. Theoretical values for the bond lengths are 0.01-0.04 A longer than the experimental ones of type r(a), in accord with general experience, but the bond angles with theoretical minus experimental differences of only 1.0-1.5 degrees are notably accurate. Symmetrized force fields were also calculated. The experimental bond lengths (r(g)/A) and angles ( 90 degree angle (alpha)()/deg) with estimated 2sigma uncertainties from GED are as follows. NbOBr(3): r(Nb=O) = 1.694(7), r(Nb-Br) = 2.429(2), 90 degree angle (O=Nb-Br) = 107.3(5), 90 degree angle (Br-Nb-Br) = 111.5(5). NbSBr(3): r(Nb=S) = 2.134(10), r(Nb-Br) = 2.408(4), 90 degree angle (S=Nb-Br) = 106.6(7), 90 degree angle (Br-Nb-Br) = 112.2(6). NbSCl(3): r(Nb=S) = 2.120(10),r(Nb-Cl) = 2.271(6), 90 degree angle (S=Nb-Cl) = 107.8(12), 90 degree angle (Cl-Nb-Cl) = 111.1(11). PMID- 12588169 TI - Novel pb and zn coordination polymers: synthesis, molecular structures, and third order nonlinear optical properties. AB - Three novel coordination polymers [Pb(bbbm)(2)(NO(3))(2)](n) (bbbm = 1,1'-(1,4 butanediyl)bis-1H-benzimidazole) 1, [Zn(bbbt)(NCS)(2)](n) (bbbt = 1,1'-(1,4 butanediyl)bis-1H-benzotriazole) 2, and [Zn(pbbt)(NCS)(2)](n) (pbbt = 1,1'-(1,3 propylene)bis-1H-benzotriazole) 3 were synthesized and structurally characterized. Polymer 1 exhibits a two-dimensional rhombohedral grid network structure, the dimensions of the grid are 14.274 x 14.274 A, and the diagonal-to diagonal distances are 24.809 x 14.125 A. Polymer 2 possesses a concavo-convex chain structure different from those of the known one-dimensional polymers, which are linear chain, zigzag chain, helical chain, double-stranded chain, and ladder chain. Polymer 3 exhibits a one-dimensional zigzag chain structure, and these chains were packed as an.ABAB. layered structure. The third-order nonlinear optical (NLO) properties of polymers1, 2, and 3 were determined with a 7-ns pulsed laser at 532 nm. 1 shows strong third-order NLO absorptive and refractive properties, and its alpha(2) and n(2) values were calculated to be 5.8 x 10(-)(9) m W(-)(1) and 4.67 x 10(-)(18) m(2) W(-)(1) in a 3.4 x 10(-)(4) mol dm(-)(3) DMF solution, respectively. Both 2 and 3 exhibit weaker NLO absorption and strong refractive properties, and their n(2) values are 4.53 x 10(-)(18) m(2) W(-)(1) for 2 in a 5.2 x 10(-)(4) mol dm(-)(3) DMF solution and 3.02 x 10(-)(18) m(2) W( )(1) for 3 in a 4.35 x 10(-)(4) mol dm(-)(3) DMF solution. The chi((3)) values of 1, 2, and 3 were calculated to be 1.67 x 10(-)(11), 1.62 x 10(-)(11), and 1.08 x 10(-)(11) esu, respectively, and the values are larger than those of the reported coordination polymers. We deduce that the valence shell structures of metal ions may have some influence on the strength of NLO properties, and discuss the relationships between the crystal structures of coordination polymers and the observed NLO properties. PMID- 12588170 TI - Metal-dependent ferro- versus antiferromagnetic interactions in molecular crystals of square Planar (M(II) imino-nitroxide radical) complexes (M = Pt, Pd). AB - The synthesis and structural, spectral, and magnetic characterizations of two new complexes of formula [Pt(IM(2)Py)Cl(2)] (A) and [Pd(IM(2)Py)Cl(2)] (B) are reported. IM(2)Py stands for the imino-nitroxide radical ligand 2-(ortho-pyridyl) 4,4,5,5-tetramethylimidazoline-1-oxyl. Their crystal structures were solved at room temperature and at 120 K revealing structural phase transitions from pseudo orthorhombic to monoclinic systems for the two compounds which remain isostructural in the whole temperature range explored. Structural parameters for A: T = 293 K [120 K], monoclinic (P2(1)/n) [P2(1)/c], a = 7.906(2) [7.989(3)] A, b = 17.872(9) [10.168(4)] A, c = 10.357(3) [17.623(6)] A, beta = 90.732(13) degrees [95.940(2)] degrees, Z = 4 [4]. Structural parameters for B: T = 293 K [120 K], monoclinic (P2(1)/n) [P2(1)/c], a = 7.900(3) [7.9730(2)] A, b = 17.907(9) [10.1806(3)] A, c = 10.299(3) [17.7171(4)] A, beta = 90.524(14) degrees [95.747(2)] degrees, Z = 4 [4]. In both complexes, the metal coordination is essentially planar. The average Pt-N, Pt-Cl and Pd-N, Pd-Cl bond lengths are 1.996(6) [1.88], 2.295(2) [2.248(8)] A and 2.015(7) [2.029(8)], 2.287(3) [2.294(3)] A, respectively. The solid state structure is characterized by a pairlike molecular packing stacked in columns parallel to the a axis; this dimer character is reinforced at low temperature. Despite their structural similarity, the investigation of the magnetic properties revealed that dominant ferromagnetic interactions govern the behavior of the Pt derivative A, whereas antiferromagnetic interactions take place for the Pd compound B. A rationalization for this rather intriguing difference is proposed in light of the spin population deduced from density functional theory calculations. The electronic absorption spectra of A and B present structured absorption bands in the visible which are attributed to MLCT transitions. Both compounds are nonluminescent at room temperature. However, a weak emission is detected for A in butyronitrile glasses at 77 K, indicating that the MLCT excited state is strongly quenched at low temperature. PMID- 12588171 TI - First example of mu(3)-sulfido bridged mixed-valent triruthenium complex triangle Ru(III)(2)Ru(II)(O,O-acetylacetonate)(3)(mu-O,O,gamma-C-acetylacetonate)(3)(mu(3) S) (1) incorporating simultaneous O,O- and gamma-C-bonded bridging acetylacetonate units. Synthesis, crystal structure, and spectral and redox properties. AB - The reaction of mononuclear ruthenium precursor [Ru(II)(acac)(2)(CH(3)CN)(2)] (acac = acetylacetonate) with the thiouracil ligand (2-thiouracil, H(2)L(1) or 6 methyl -2-thiouracil, H(2)L(2)) in the presence of NEt(3) as base in ethanol solvent afforded a trinuclear triangular complex Ru(3)(O,O-acetylacetonate)(3)(mu O,O,gamma-C-acetylacetonate)(3)(mu(3)-sulfido) (1). In 1, each ruthenium center is linked to one usual O,O-bonded terminal acetylacetonate molecule whereas the other three acetylacetonate units act as bridging functions: each bridges two adjacent ruthenium ions through the terminal O,O-donor centers at one end and via the gamma-carbon center at the other end. Moreover, there is a mu(3)-sulfido bridging in the center of the complex unit, which essentially resulted via the selective cleavage of the carbon-sulfur bond of the thiouracil ligand. In diamagnetic complex 1, the ruthenium ions are in mixed valent Ru(III)Ru(III)Ru(II) state, where the paramagnetic ruthenium(III) ions are antiferromagnetically coupled. The single crystal X-ray structure of 1 showed two crystallographically independent C(3)-symmetric molecules, Ru(3)(O,O acetylacetonate)(3)(mu-O,O,gamma-C-acetylacetonate)(3)(mu(3)-S) (1), in the asymmetric unit. Bond distances of both crystallographically independent molecules are almost identical, but there are some significant differences in bond angles (up to 6 degrees ) and interplanar angles (up to 8 degrees ). Each ruthenium atom exhibits a distorted octahedral environment formed by four oxygen atoms, two from each of the terminal and bridging acetylacetonate units, one gamma-carbon of an adjacent acetylacetonate ligand, and the sulfur atom in the center of the complex. In agreement with the expected 3-fold symmetry of the complex molecule, the (1)H and (13)C NMR spectra of 1 in CDCl(3) displayed signals corresponding to two types of ligand units. In dichloromethane solvent, 1 exhibited three metal center based successive quasireversible redox processes, Ru(III)Ru(III)Ru(III)-Ru(III)Ru(III)Ru(II) (couple I, 0.43 V vs SCE); Ru(III)Ru(III)Ru(IV)-Ru(III)Ru(III)Ru(III) (couple II, 1.12 V); and Ru(III)Ru(III)Ru(II)-Ru(III)Ru(II)Ru(II) (couple III, -1.21 V). However, in acetonitrile solvent, in addition to the three described couples [(couple I), 0.34 V; (couple II), 1.0 V; (couple III), -1.0], one irreversible oxidative response (Ru(III)Ru(III)Ru(IV) --> Ru(III)Ru(IV)Ru(IV) or oxidation of the coordinated sulfide center) appeared at E(pa), 1.50 V. The large differences in potentials between the successive couples are indicative of strong coupling between the ruthenium ions in the mixed-valent states. Compound 1 exhibited a moderately strong charge-transfer (CT) transition at 654 nm and multiple ligand based intense transitions in the UV region. In the Ru(III)Ru(III)Ru(III) (1(+)) state, the CT band was slightly blue shifted to 644 nm; however, the CT band was further blue shifted to 520 nm on two-electron oxidation to the Ru(III)Ru(III)Ru(IV) (1(2+)) state with a reduction in intensity. PMID- 12588172 TI - Germanium-rich pentaruthenium carbonyl clusters including Ru(5)(CO)(11)(mu GePh(2))(4)(mu(5)-C) and its reactions with hydrogen. AB - The reaction of Ru(5)(CO)(15)(mu(5)-C), 1, with Ph(3)GeH at 150 degrees C has yielded two new germanium-rich pentaruthenium cluster complexes: Ru(5)(CO)(11)(mu CO)(mu-GePh(2))(3)(mu(5)-C), 2; Ru(5)(CO)(11)(mu;-GePh(2))(4)(mu(5)-C), 3. Both compounds contain square pyramidal Ru(5) clusters with GePh(2) groups bridging three and four of the edges of the Ru(5) square base, respectively. When treated with 1 equiv of Ph(3)GeH at 150 degrees C compound 2 is converted to 3. Reaction of 3 with H(2) at 150 degrees C yielded Ru(5)(CO)(10)(mu-GePh(2))(4)(mu(5)-C)(mu H)(2), 4, containing two hydride ligands and one less CO ligand. Reaction of 4 with hydrogen at 150 degrees C yielded the compound Ru(5)(CO)(10)(mu GePh(2))(2)(mu(3)-GePh)(2)(mu(3)-H)(mu(4)-CH), 5, by loss of benzene and conversion of two of the bridging GePh(2) groups into triply bridging GePh groups. Compound 5 contains one triply bridging hydride ligand and a quadruply bridging methylidyne ligand formed by addition of one hydrogen atom to the carbido carbon atom. PMID- 12588173 TI - Stability of the gold(i)-phosphine bond. A comparison with other group 11 elements. AB - The stability of gold phosphine complexes of the form [Au(PH(3))(n)()](+) (n = 1 4) and [AuCl(PH(3))(n)()] (n = 1-3) is analyzed in detail by applying quantum theoretical methods and compared to the coordination behavior of the lighter group 11 elements copper and silver. It is shown that, once [M(PH(3))(2)](+) or [MClPH(3)] (M = Cu, Ag, and Au) is formed, further coordination by PH(3) ligands is relatively weak; i.e., the energy gain to form [M(PH(3))(3)](+) from [M(PH(3))(2)](+) is less than 60 kJ mol(-)(1), and less than 100 kJ mol(-)(1) to form [MCl(PH(3))(2)] from [MClPH(3)]. Relativistic effects in gold significantly influence these factors and reduce the tendency for phosphine coordination beyond two-coordination. This implies that the most favored coordination number for gold is two with either a linear P-Au-P or P-Au-X arrangement (X = a strongly coordinating ligand like Cl(-)). Instead, X-Au-PH(3) units prefer to interact via close Au-Au contacts (aurophilic interactions) keeping the linear structure approximately intact, while the corresponding copper and silver compounds prefer PH(3) coordination to strongly bound M(2)Cl(2) units (M = Cu or Ag) where two chlorine atoms bridge the two metal atoms thus having the formal coordination number of three for copper or silver. PMID- 12588174 TI - Aqua Ions. 1. The structures of the [Ru(OH(2))(6)](3+) and [V(OH(2))(6)](3+) cations in aqueous solution: an EPR and UV-Vis study. AB - Spectroscopic data are presented for the [V(OH(2))(6)](3+) and [Ru(OH(2))(6)](3+) cations, from which inferences are drawn regarding their structures in aqueous solution. EPR and absorption spectra of solutions and glasses are supplemented by spectra of the aqua ions in various crystalline environments, and the electronic and molecular structures inter-related through elementary angular overlap model calculations. It is concluded that in aqueous solution the [Ru(OH(2))(6)](3+) cation is localized in the all-horizontal D(3)(d)geometry, whereas the structure of the [V(OH(2))(6)](3+) cation is close to T(h) symmetry. These results are consistent with the most energetically favored geometries predicted by ab initio calculations. PMID- 12588175 TI - Aqua Ions. 2. Structural manifestations of the Jahn-Teller effect in the beta alums. AB - Variable-temperature single-crystal neutron diffraction structures of the alums CsM(III)(SO(4))(2).12D(2)O, where M(III) = Ti, V, Mn, and Ga, are reported. Structural differences are highlighted by the titanium and manganese alums, which undergo cubic (Pathremacr;) to orthorhombic (Pbca) phase transitions at approximately 13 and approximately 156 K, respectively. The structural instability exhibited by these salts is interpreted as arising from cooperative Jahn-Teller interactions, and these measurements characterize the structural changes that result from the coupling between the electronic and vibrational states. Although the symmetry changes associated with the phase transformations are analogous for the Ti and Mn alums, the low-temperature geometries of the tervalent hexaaqua cations are markedly different. Whereas the MnO(6) framework is subject to a pronounced tetragonal elongation, changes in the Ti-O bond lengths are very modest; but significant changes in the O-Ti-O bond angles and in the disposition of the coordinated water molecules are identified. The large differences in the transition temperatures and in the low-temperature stereochemistries of the [Ti(OD(2))(6)](3+) and [Mn(OD(2))(6)](3+) cations are related to the sensitivity of the energies of the t(2g) (O(h)) and e(g) (O(h)) orbitals to the various asymmetric vibrations of the hexaaqua complex. PMID- 12588176 TI - K(2)MM'(3)Se(6) (M = Cu, Ag; M' = Ga, In), A new series of metal chalcogenides with chain-sublayer-chain slabs: (infinity)(1)[M'Se(4)] (infinity)(2)[(MSe(4))(M'Se(4))]-(infinity)(1)[M'Se(4)]. AB - A new series of novel isostructural metal chalcogenides, K(2)CuIn(3)Se(6) (1), K(2)CuGa(3)Se(6) (2), and K(2)AgIn(3)Se(6) (3), were obtained by a reactive flux technique and structurally characterized. Compounds 1, 2, and 3 crystallize in the space group C2/c of the monoclinic system with eight formula units in a cell: a = 11.445(2) A, b = 11.495(2) A, c = 21.263(4) A, beta = 97.68(3) degrees, V = 2772(1) A(3), R1/wR2 = 0.0676/0.1652 for 1; a = 11.031(2) A, b = 11.050(4) A, c = 20.808(7) A, beta = 97.71(2) degrees, V = 2513(1) A(3), R1/wR2 = 0.0301/0.0511 for 2; and a = 11.633(1) A, b = 11.587(1) A, c = 21.355(1) A, beta = 98.010(8) degrees, V = 2850.4(4) A(3), R1/wR2 = 0.0471/0.0732 for 3. These isostructural compounds are characterized by a chain-sublayer-chain slab structure. The sublayer, composed of alternative corner-sharing mixed-metal tetrahedra, is sandwiched by parallel corner-sharing tetrahedral chains. Optical absorption spectra of compounds 1, 2, and 3 reveal the presence of a sharp optical gap of 1.68, 1.72, and 1.64 eV, respectively, suggesting that these materials are semiconductors and suitable for efficient absorption of solar radiation in solar cell applications. IR spectra show no obvious absorption in the range 800-4000 cm(-)(1). PMID- 12588177 TI - Redox active macrocyclic receptors for neutral guests. AB - A novel series of triazine-appended macrocyclic complexes has been investigated as potential hydrogen bonding receptors for complementarily disposed heterocycles. Cocrystallization of a melamine-appended azacyclam complex of Cu(II) has been achieved with barbitone, the barbiturate anion and thymine. In each case, a complementary DAD/ADA hydrogen bonding motif between the melamine group and the heterocycle has been identified by X-ray crystallography. Electrochemical studies of the copper macrocycles in both nonaqueous and aqueous solution show anodic shifts of the Cu(II/)(I) redox couple of more than 60 mV upon addition of guest molecules with matching H-bonding motifs. The Zn(II) analogues have been synthesized via transmetalation of the Cu(II) complex, and their guest binding properties investigated by NMR spectroscopy. (1)H NMR shifts of up to 0.8 ppm were observed upon addition of guest, and stability constants are similar to those obtained electrochemically. PMID- 12588178 TI - Enantiospecific synthesis of delta and lambda [Ru(bpy)(2)ppy](+) and [Ru(bpy)(2)quo](+) (bpy = 2,2'-bipyridine, ppy = phenylpyridine-H(+), quo = 8 hydroxyquinolate): (1)H and (13)C NMR studies and X-ray structure determination of rac-[Ru(bpy)(2)quo]PF(6). AB - In this paper, we describe the enantiospecific synthesis and the complete characterization of the two hexacoordinated ruthenium(II) monocations [Ru(bpy)(2)ppy](+) and [Ru(bpy)(2)quo](+) (bpy = 2,2'-bipyridine, ppy = phenylpyridine-H(+), quo = 8-hydroxyquinolate) in their enantiomeric Delta and Lambda forms. The corresponding enantiomeric excesses (ee's) are determined by (1)H NMR using pure Delta-Trisphat (tris(tetrachlorobenzenedialato)phosphate(V) anion) as a chiral (1)H NMR shift reagent. A complete (1)H and (13)C NMR study has been carried out on rac-[Ru(bpy)(2)ppy]PF(6) and rac-[Ru(bpy)(2)quo]PF(6). Additionally, the X-ray molecular structure of rac-[Ru(bpy)(2)quo]PF(6) is reported; this latter species crystallizes in the monoclinic C2/c space group (a = 22.079 A, b = 16.874 A, c = 17.533 A, alpha = 90 degrees, beta = 109.08 degrees, gamma = 90 degrees ). PMID- 12588180 TI - Analysis of cyanobacterial toxins by immunological methods. PMID- 12588181 TI - Mechanism of ascididemin-induced cytotoxicity. AB - Some marine animals are rich sources of unique polycyclic aromatic alkaloids that are cytotoxic against tumor cell lines and effective in mouse tumor xenograft models. Ascididemin is a pyridoacridine alkaloid originally derived from a Didemnum sp. tunicate. It has potent cytotoxicity against tumor cells in vitro and in vivo. Preclinical screening at NCI revealed the antineoplastic activities of ascididemin and a synthetic analogue 48. Ascididemin has been reported to inhibit topoisomerase II and induce topoisomerase II-mediated DNA cleavage. This study, however, focuses on the unique ability of ascididemin and two synthetic analogues (48 and 109) to cleave DNA in the absence of topoisomerase I or II. An in vitro assay revealed their concentration-dependent ability to cleave DNA and identified dithiothreitol as the sole requirement for maximal activity. On the basis of shared structural features of the three analogues, a double N-bay region and iminoquinone heterocyclic ring, two possible mechanisms of action were hypothesized: (1) generation of reactive oxygen species facilitated by metal binding to the common phenanthroline bay region, and (2) production of reactive oxygen species by direct reduction of the iminoquinone moiety. Experimental results supported direct iminoquinone reduction and ROS generation as the mechanism of ascididemin cytotoxicity. Antioxidants protected against DNA cleavage in vitro and protected cultured Chinese hamster ovary cells from toxicity. Additionally, it was shown that cells deficient in the ability to repair reactive oxygen species damage to their DNA were more susceptible to ascididemin and analogues than repair competent cells. Ascididemin-treated cells were also shown to induce oxygen-stress related proteins, further implicating the production of reactive oxygen species as the mechanism of cytotoxicity for these molecules. PMID- 12588182 TI - In vitro metabolism of tolcapone to reactive intermediates: relevance to tolcapone liver toxicity. AB - Tolcapone is a catechol-O-methyltransferase (COMT) inhibitor used for control of motor fluctuations in Parkinson's disease (PD). Since its entry onto the market in 1998, tolcapone has been associated with numerous cases of hepatotoxicity, including three cases of fatal fulminant hepatic failure. The cause of this toxicity is not known; however, it does not occur with the use of the structurally similar drug entacapone. It is known that tolcapone is metabolized to amine (M1) and acetylamine (M2) metabolites in humans, but that the analogous metabolites were not detected in a limited human study of entacapone metabolism. We hypothesized that one or both of these tolcapone metabolites could be oxidized to reactive species and that these reactive metabolites might play a role in tolcapone-induced hepatocellular injury. To investigate this possibility, we examined the ability of M1 and M2 to undergo in vitro bioactivation by electrochemical and enzymatic methods. Electrochemical experiments revealed that M1 and M2 are more easily oxidized than the parent compound, in the order M1 > M2 > tolcapone. There was a general correlation between oxidation potential and the half-lives of the compounds in the presence of two oxidizing systems, horseradish peroxidase and myeloperoxidase. These enzymes catalyzed the oxidation of M1 and M2 to reactive species that could be trapped with glutathione (GSH) to form metabolite adducts (C1 and C2). Each metabolite was found to only form one GSH conjugate, and the structures were tentatively identified using LC-MS/MS. Following incubation of M1 and M2 with human liver microsomes in the presence of GSH, the same adducts were observed, and their structures were confirmed using LC MS/MS and (1)H NMR. Experiments with chemical P450 inhibitors and cDNA-expressed P450 enzymes revealed that this oxidation is catalyzed by several P450s, and that P450 2E1 and 1A2 play the primary role in the formation of C1 while P450 1A2 is most important for the production of C2. Taken together, these data provide evidence that tolcapone-induced hepatotoxicity may be mediated through the oxidation of the known urinary metabolites M1 and M2 to reactive intermediates. These reactive species may form covalent adducts to hepatic proteins, resulting in damage to liver tissues, although this supposition was not investigated in this study. PMID- 12588184 TI - Computational and experimental studies on the distribution of addition and substitution products of the microsomal glutathione transferase 1-catalyzed conjugation of glutathione with fluoroalkenes. AB - The glutathione transferase-catalyzed reaction of glutathione with haloalkenes results in the formation of addition or substitution products or both. Glutathione conjugates of haloalkenes may be metabolized and excreted at different rates, may follow different metabolic pathways, and may exhibit different toxicities. Microsomal glutathione transferase 1 (MGST1)-catalyzed conjugation of chlorotrifluoroethene, hexafluoropropene, and 2-(fluoromethoxy) 1,1,3,3,3-pentafluoro-1-propene results in differing proportions of addition and substitution products. The aim of the present study was to develop a computational model to predict the outcome of the MGST1-catalyzed reaction of glutathione with haloalkenes. An ab initio computational study of the reaction of ethanethiolate, a surrogate for glutathione, with the chlorotrifluoroethene, hexafluoropropene, and 2-(fluoromethoxy)-1,1,3,3,3-pentafluoro-1-propene was conducted. An empirical study was also conducted to quantify the distribution of addition and substitution products that resulted from the MGST1-catalyzed reaction of glutathione with these fluoroalkenes. The results show that this computational model accurately predicted the distribution of the addition and substitution products that result from the MGST1-catalyzed reaction of glutathione with these fluoroalkenes. PMID- 12588183 TI - Mutation of tyrosine 190 to alanine eliminates the inactivation of cytochrome P450 2B1 by peroxynitrite. AB - We have previously reported that cytochrome P450 2B1 was inactivated by peroxynitrite and that the decrease in the catalytic activity correlated with an increase in the nitration of tyrosine. Digestion of the peroxynitrite-treated P450 2B1 with Lys C followed by amino acid sequencing of the major nitrotyrosine containing peptide demonstrated that it spanned residues 160-225. This peptide contains two tyrosine residues at positions 190 and 203. In this study, we mutated Tyr 190 to Ala (Y190A) and Tyr 203 to Ala (Y203A) in wild-type recombinant P450 2B1 (WT) in order to identify the specific residue(s) that is nitrated and to determine whether nitrotyrosine formation is reponsible for the peroxynitrite-mediated inactivation of P450 2B1. All three P450s were expressed in Escherichia coli, purified to homogeneity, and characterized. The catalytic activities for four different substrates of P450 2B1 increased approximately 2 fold for the Y203A mutant, but decreased by about 60% for the Y190A mutant when compared to WT. The addition of peroxynitrite to the P450s resulted in concentration-dependent decreases in the catalytic activities of WT and Y203A, but no loss of the catalytic activities of Y190A. The extent of tyrosine nitration of Y190A by peroxynitrite decreased by approximately 75% as compared with WT or the Y203A protein. Following digestion of the peroxynitrite-modified proteins with Lys C, a major nitrotyrosine-containing peptide was detected from WT and Y203A, but not from Y190A. Collectively, these results indicate that Tyr 190 is the target residue for peroxynitrite-mediated nitration and that nitration of this tyrosine is a responsible for the inactivation of P450 2B1. Modeling studies suggest that Tyr 190 may play a structural role in maintaining the integrity of the protein for maximal activity through hydrogen bonding with Glu 149. PMID- 12588185 TI - Reactions of formaldehyde plus acetaldehyde with deoxyguanosine and DNA: formation of cyclic deoxyguanosine adducts and formaldehyde cross-links. AB - We investigated the reactions of formaldehyde plus acetaldehyde with dGuo and DNA in order to determine whether certain 1,N(2)-propano-dGuo adducts could be formed. These adducts-3-(2'-deoxyribosyl)-5,6,7,8-tetrahydro-8 hydroxypyrimido[1,2-a]purine-(3H)-one (1) and 3-(2'-deoxyribosyl)-5,6,7,8 tetrahydro-6-hydroxypyrimido[1,2-a]purine-(3H)-one (3a,b)-have been previously characterized as products of the reaction of acrolein with dGuo and DNA. Adduct 1 predominates in certain model lipid peroxidation systems [Pan, J., and Chung, F. L. (2002) Chem. Res. Toxicol. 15, 367-372]. We hypothesized that this could be due to stepwise reactions of formaldehyde and acetaldehyde with dGuo, rather than by reaction of acrolein with dGuo. The results demonstrated that adducts 1 and 3a,b were relatively minor products of the reaction of formaldehyde and acetaldehyde with dGuo and that there was no selectivity in their formation. These findings did not support our hypothesis. However, substantial amounts of previously unknown cyclic dGuo adducts were identified in this reaction. The new adducts were characterized by their MS, UV, and NMR spectra as diastereomers of 3 (2'-deoxyribosyl)-6-methyl-1,3,5-diazinan[4,5-a]purin-10(3H)-one (10a,b). Adducts 10a,b were apparently formed by addition of formaldehyde to N1 of N(2)-ethylidene dGuo, followed by cyclization. An analogous set of four diastereomers of 3-(2' deoxyribosyl)-6,8-dimethyl-1,3,5-diazinan[4,5-a]purin-10(3H)-one (12a-d) were formed in the reactions of acetaldehyde with dGuo. These products are the first examples of exocyclic dGuo adducts of the pyrimido[1,2-a]purine type in which an oxygen atom is incorporated into the exocyclic ring. Formaldehyde-derived adducts were the other major products of the reactions of formaldehyde plus acetaldehyde with dGuo. Prominent among these were N(2)-hydroxymethyl-dGuo (9) and the cross link di-(N(2)-deoxyguaonosyl)methane (13). We did not detect adducts 1, 3a,b, or 10a,b in enzymatic hydrolysates of DNA that had been allowed to react with formaldehyde plus acetaldehyde. However, we did detect substantial amounts of the formaldehyde cross-links di-(N(6)-deoxyadenosyl)methane (17), with lesser quantities of (N(6)-deoxyadenosyl-N(2)-deoxyguanosyl)methane (18), di-(N(2) deoxyguanosyl)methane (13), and N(6)-hydroxymethyl-dAdo (19). Schiff base adducts of formaldehyde and acetaldehyde were also detected in these reactions. These results demonstrate that the reactions of formaldehyde plus acetaldehyde with dGuo are dominated by newly identified cyclic adducts and formaldehyde-derived products whereas the reactions with DNA result in the formation of formaldehyde cross-link adducts. The carcinogens formaldehdye and acetaldehyde occur in considerable quantities in the human body and in the environment. Therefore, further research is required to determine whether the adducts described here are formed in animals or humans exposed to these agents. PMID- 12588186 TI - Development of binary classification of structural chromosome aberrations for a diverse set of organic compounds from molecular structure. AB - Classification models are generated to predict in vitro cytogenetic results for a diverse set of 383 organic compounds. Both k-nearest neighbor and support vector machine models are developed. They are based on calculated molecular structure descriptors. Endpoints used are the labels clastogenic or nonclastogenic according to an in vitro chromosomal aberration assay with Chinese hamster lung cells. Compounds that were tested with both a 24 and 48 h exposure are included. Each compound is represented by calculated molecular structure descriptors encoding the topological, electronic, geometrical, or polar surface area aspects of the structure. Subsets of informative descriptors are identified with genetic algorithm feature selection coupled to the appropriate classification algorithm. The overall classification success rate for a k-nearest neighbor classifier built with just six topological descriptors is 81.2% for the training set and 86.5% for an external prediction set. The overall classification success rate for a three descriptor support vector machine model is 99.7% for the training set, 92.1% for the cross-validation set, and 83.8% for an external prediction set. PMID- 12588187 TI - Photochemical and photobiological studies of tirapazamine (SR 4233) and related quinoxaline 1,4-Di-N-oxide analogues. AB - Tirapazamine, 3-amino-1,2,4-benzotriazine 1,4-di-N-oxide (TPZ; SR 4233), is currently undergoing phase II and III clinical trials as an antitumor agent. We have studied the photochemical properties of TPZ, and the related analogues 3 amino-2-quinoxalinecarbonitrile 1,4-di-N-oxide (TPZCN) and quinoxaline-1,4-di-N oxide (quindoxin) with respect to their potential to photodamage DNA both oxidatively and reductively. We have found that TPZ, TPZCN, and quindoxin photosensitized the generation of singlet oxygen with quantum yields of 0.007, 0.19, and 0.02, respectively, in acetonitrile. Irradiation (lambda > 300 nm) of TPZ at pH 9.4 in the presence of a reducing agent, NADH, generated the corresponding nitroxide radical. At pH 7.4, photoirradiation of either TPZ or TPZCN in the presence of NADH in air saturated buffer gave the superoxide radical, which was trapped by 5,5-dimethyl-1-pyrroline N-oxide (DMPO). In the absence of a reducing agent, singlet oxygen generated from TPZCN oxidized DMPO to 5,5-dimethyl-2-oxopyrrolin-1-oxyl (DMPOX). No spin adducts were detected during photoirradiation of TPZ, NADH, and DMPO in nitrogen-saturated buffer. However, when DMSO was also present, the DMPO/(*)CH(3) adduct was observed, indicating the generation of the free hydroxyl radical. Both TPZ and TPZCN photooxidized reduced glutathione and azide to the glutathiyl and azidyl radicals, respectively. Under anaerobic conditions, NADH increased photoinduced strand breaks in pBR322 plasmid DNA caused by TPZ or TPZCN. For TPZ, the reactive species is probably the aforementioned nitroxide radical or the hydroxyl radical generated from its decomposition. In contrast, DNA damage by quindoxin was not affected by NADH, suggesting a different mechanism, possibly involving a photogenerated oxaziridine intermediate. These studies show that the photochemistry of TPZ, TPZCN, and quindoxin is complex and depends on the redox environment and whether oxygen is present. PMID- 12588188 TI - Molecular characterization of mitomycin C-induced large deletions and tandem-base substitutions in the bone marrow of gpt delta transgenic mice. AB - Deletion mutations constitute an important class of mutations that may result in a variety of human diseases, including cancer. Although many chemicals and ionizing radiations induce deletions, this class of mutation has been poorly characterized at the molecular level, particularly in vivo. Here we report the molecular nature of deletions as well as base substitutions induced by antitumor antibiotic mitomycin C (MMC) in the bone marrow using a novel transgenic mouse, gpt delta. In this mouse model, deletions and point mutations in lambda DNA integrated in the chromosome are individually selected as Spi(-) (sensitive to P2 interference) phages and 6-thioguanine-resistant bacterial colonies, respectively. The mice were treated with MMC (1 mg/kg/day) for five consecutive days. One week after the last treatment, lambda phage was rescued from the genomic DNA of the bone marrow by in vitro packaging reactions and subjected to Spi(-) and 6-thioguanine selections. The mutant frequency of Spi(-) with large deletions increased more than 20-fold over that of the control. Molecular sizes of the large deletions were mostly more than 2,000 base pairs. The large deletions frequently occurred between two short direct repeat sequences from 2 to 6 base pairs, suggesting that they are generated during the end-joining repair of double-strand breaks induced by interstrand cross-links in DNA. In 6-thioguanine selection, tandem-base substitutions, such as 5'-GG-3' to 5'-AT-3', were induced. It highlights the relevance of intrastrand cross-links as genotoxic lesions. Previous in vitro studies report the induction of single-base substitutions and single-base deletions by MMC. However, no such mutations were identified in vivo. Thus, our results strongly caution that in vitro mutation spectra do not necessarily reflect genotoxic events in vivo and emphasize the importance of transgenic rodent genotoxicity assays to examine the roles of DNA adducts in mutagenesis and carcinogenesis. PMID- 12588189 TI - Identification of adducts produced by the reaction of 4 (acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanol with deoxyguanosine and DNA. AB - 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is a metabolite of the tobacco specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). NNAL is present in the blood and urine of people exposed to tobacco products and has carcinogenic activity in rodents similar to that of NNK. DNA adducts specific to NNAL have not been previously identified. Metabolic activation of NNAL by alpha-methyl hydroxylation, a pathway known to occur in rodent and human microsomes, would produce pyridylhydroxybutylating agents that could react with DNA. We investigated this possibility in the present study by allowing 4 (acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone (NNALCH(2)OAc) to react with dGuo and DNA. Products were identified by HPLC with UV detection, liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS) and LC/ESI tandem mass spectrometry (LC/ESI-MS/MS). In the dGuo reactions, selected ion monitoring for m/z 417, corresponding to pyridylhydroxybutylated dGuo, showed several peaks. One adduct was identified as 7-[1-hydroxy-1-(3-pyridyl)but-4 yl]dGuo (21) by neutral thermal hydrolysis, which converted it to 7-[1-hydroxy-1 (3-pyridyl)but-4-yl]Gua (22) and 4-hydroxy-1-(3-pyridyl)-1-butanol (16). Adduct 22 was identified by comparison of its LC/ESI-MS and LC/ESI-MS/MS properties to those of standard 22. Two other adducts, O(6)-[1-hydroxy-1-(3-pyridyl)but-4 yl]dGuo (17) and N(2)-[1-hydroxy-1-(3-pyridyl)but-4-yl]dGuo (19), were identified by comparison of their LC/ESI-MS and LC/ESI-MS/MS properties to those of standard 17 and 19. Further evidence for the identity of 17 and 19 was obtained by mild acid hydrolysis, which converted them to the corresponding Gua bases 18 and 20, identified by comparison to synthetic standards. Neutral thermal hydrolysis of DNA that had been reacted with NNALCH(2)OAc produced 22, identified by comparison to a standard. Adducts 17 and 19 were identified in enzyme hydrolysates of this DNA by comparison to standards. Thus, DNA that had been allowed to react with NNALCH(2)OAc contained adducts 17, 19, and 21. The results of this study provide markers for investigating the role of specific NNAL-DNA adducts in carcinogenesis by NNAL and NNK. PMID- 12588190 TI - Hydroxyl radical is not the main reactive species involved in the degradation of DNA bases by copper in the presence of hydrogen peroxide. AB - Copper is an important biological metal that tightly binds to DNA. Its reaction with endogenously generated hydrogen peroxide may thus lead to the formation of DNA damage. To gain insights into the underlying mechanisms, a comparative study of the damage produced within isolated DNA upon exposure to gamma-radiation in aqueous solution, a source of hydroxyl radicals, and incubation with Cu(I) or Cu(II) complexes in the presence of hydrogen peroxide was carried out. Several relevant base modifications were quantified by HPLC-tandem mass spectrometry. It was first shown that addition of copper ions only slightly modified the profile of radiation-induced lesions within DNA. However, the distribution of base modifications was drastically different upon incubation of DNA with Cu(I) or Cu(II) complexes in the presence of H(2)O(2). Indeed, guanine degradation products were produced in much higher yield than lesions of the other bases. These observations are rationalized in terms of the occurrence of one electron oxidation with Cu(I) complexes, as confirmed by the study of the degradation of free thymidine. In contrast, the formation of the sole 8-oxo-7,8-dihydroguanine upon incubation of DNA with Cu(II) ions and H(2)O(2) strongly suggests the production of singlet oxygen as the predominant reactive oxygen species. PMID- 12588191 TI - Metabolic activation of a pyrazinone-containing thrombin inhibitor. Evidence for novel biotransformation involving pyrazinone ring oxidation, rearrangement, and covalent binding to proteins. AB - Compound I, (2-[3-[(2,2-difluoro-2(2-pyridyl)ethyl)amino]-6-methyl-2 oxohydropyrazinyl]-N-[(3-fluoro(2-pyridyl))methyl]acetamide, is a potent competitive inhibitor of thrombin that reacts stoichiometrically with the protease. Compounds of this class possess therapeutic potential as anticoagulation agents. During the metabolic characterization of compound I, evidence was obtained for extensive metabolic activation of the pyrazinone ring system. Following administration of (14)C-labeled I to rats, significant levels of irreversibly bound radioactivity to proteins were detected in rat plasma and liver. LC/MS/MS analysis of metabolites formed in rat and human liver microsomes fortified with glutathione (GSH) revealed the presence of two structurally distinct GSH adducts. It is proposed that the first of these GSH conjugates derives from a two electron oxidation of the 6-methyl-2-oxo-3-aminopyrazinone moiety to afford an electrophilic imine-methide intermediate, while the second is formed by addition of GSH to an epoxide formed by P-450-mediated oxidation of the double bond at the 5-6 position of the pyrazinone ring. The addition of GSH to the proposed epoxide facilitates opening of the pyrazinone ring and a rearrangement to afford a stable, rearranged imidazole-containing metabolite. Elucidation of the metabolic activation pathways of I provides structural guidance for the design of thrombin inhibitors with decreased potential for the generation of chemically reactive intermediates. PMID- 12588192 TI - Reactivity of 1,4-dihydropyridines toward alkyl, alkylperoxyl radicals, and ABTS radical cation. AB - A series of C4-substituted 1,4-dihydropyridines (DHP) with either secondary or tertiary nitrogen in the dihydropyridine ring were synthesized. All of these compounds together with some commercial DHP derivatives were tested for potential scavenger effects toward alkyl, alkylperoxyl radicals, and ABTS radical cation in aqueous media at pH 7.4. Kinetic rate constants were assessed either by UV/vis spectroscopy or GC/MS techniques. Tested compounds reacted faster toward alkylperoxyl radicals and ABTS radical cation than alkyl ones. N-Ethyl substituted DHPs showed the lowest reactivity. Kinetic results were compared with either trolox or nisoldipine. Using deuterium kinetic isotope effect studies, we have proved that the hydrogen of the 1-position of the DHP ring is involved in the proposed mechanism. This fact is mostly noticeable in the case of alkyl radicals. In all cases, the respective pyridine derivative was detected as the main product of the reaction. PMID- 12588193 TI - Glutathione transferase theta 1-1-dependent metabolism of the water disinfection byproduct bromodichloromethane. AB - Bromodichloromethane (CHBrCl(2)), a prevalent drinking water disinfection byproduct, was previously shown to be mutagenic in Salmonella that express rat GSH transferase (GST) theta 1-1 (GST T1-1). In the present study, in vitro experiments were performed to study the kinetics of CHBrCl(2) reactions mediated by GST in different species as well as the isoform specificity and reaction products of the GST pathway. Conjugation activity of CHBrCl(2) with GSH in mouse liver cytosol was time- and protein-dependent, was not inhibited by the GST alpha, mu and pi inhibitor S-hexyl-GSH, and correlated with GST T1-1 activity toward the substrate 1,2-epoxy-3-(4'-nitrophenoxy)propane. Conjugation activities in hepatic cytosols of different species toward CHBrCl(2) followed the order mouse > rat > human. As compared with CH(2)Cl(2), the catalytic efficiency (k(cat)/K(m)) of conjugation of CHBrCl(2) with GSH by pure recombinant rat GST T1 1 was approximately 3-6-fold less. Taken together, this suggests that GST T1-1 is the primary catalyst for conjugation of CHBrCl(2) with GSH and that flux through this pathway is less than for CH(2)Cl(2). The initial GSCHCl(2) conjugate formed was unstable and degraded to several metabolites, including GSCH(2)OH, S-formyl GSH, and HCOOH. Addition of NAD(+) to cytosol did not alter the rate of conjugation of CHBrCl(2) with GSH; however, it did increase the amount of [(14)C]HCOOH produced ( approximately 10-fold). A similar result was seen in a reaction containing pure rat GST T1-1 and GSH-dependent formaldehyde dehydrogenase, indicating that GSCH(2)OH was formed as a precursor to S-formyl GSH. The half-life of synthetic S-formyl-GSH in pH 7.4 buffer was approximately 1 h at ambient temperature and decreased to approximately 7 min in pH 9.0 buffer, and it does not react with deoxyguanosine. In conclusion, GST T1-1 conjugation of CHBrCl(2) has been definitively demonstrated and the kinetics of conjugation of CHBrCl(2) with GSH characterized in mouse, rat, and human hepatic cytosols. The significance of this GST pathway is that reactive GSH conjugates are produced resulting in possible formation of DNA adducts. Comparisons with CH(2)Cl(2) suggest that the reactive intermediates specific to GSH conjugation of CHBrCl(2) are more mutagenic/genotoxic than those derived from CH(2)Cl(2). PMID- 12588194 TI - Metabolism of benzo[c]chrysene and comparative mammary gland tumorigenesis of benzo[c]chrysene bay and fjord region diol epoxides in female CD rats. AB - Benzo[c]chrysene (BcC), an environmental pollutant, is a unique polycyclic aromatic hydrocarbon that possesses both a bay region and a fjord region in the same molecule. We previously demonstrated that both bay region and fjord region terminal rings are involved in the in vitro metabolism of BcC. In the present investigation, we prepared [14-(3)H]BcC and tested the hypothesis that BcC can be activated to both bay region and fjord region diol epoxides in female CD rats. At 6 weeks of age, rats were gavaged with a single dose of [14-(3)H]BcC (5 mg/rat; specific activity, 6.7 Ci/mmol) in 0.5 mL of trioctanoin. During the first 48 h, 20.3% of the dose was eliminated in the feces and 2.8% was eliminated in the urine. After 1 week, cumulatively, 23.2 and 3.5%, respectively, were eliminated. 3-Hydroxybenzo[c]chrysene, 10-hydroxybenzo[c]chrysene, and trans-7,8-dihydroxy 7,8-dihydrobenzo[c]chrysene were the major fecal metabolites. In urine, trans-1,2 dihydroxy-1,2-dihydrobenzo[c]chrysene, 2-hydroxybenzo[c]chrysene, (+/-)-1,t-2,t 3,c-4-tetrahydroxy-1,2,3,4-tetrahydrobenzo[c]chrysene, and (+/-)-9,t-10,t-11,c-12 tetrahydroxy-9,10,11,12-tetrahydrobenzo[c]chrysene were detected, primarily as glucuronic acid and sulfate conjugates. The identification of the two tetraols clearly indicates that both bay region and fjord region diol epoxides are formed as intermediates in the metabolism of BcC in vivo. The second goal of this study was to test the hypothesis that the location of the epoxide moiety (fjord vs bay region) determines the carcinogenic activity. Thus, we compared the carcinogenicity of the bay region (+/-)-anti-1,2-dihydroxy-3,4-epoxy-1,2,3,4 tetrahydrobenzo[c]chrysene and the fjord region (+/-)-anti-9,10-dihydroxy-11,12 epoxy-9,10,11,12-tetrahydrobenzo[c]chrysene in the rat mammary gland. The results clearly showed that the fjord region diol epoxide is a potent mammary carcinogen, while the bay region diol epoxide lacks activity in this model assay. This is the first report on a comparison of mammary cancer induction by fjord and bay region diol epoxides derived from the same molecule. It further supports previous observations that fjord region diol epoxides are more carcinogenic than structurally related bay region diol epoxides. PMID- 12588195 TI - Model studies on the modification of proteins by lipoxidation-derived 2 hydroxyaldehydes. AB - 2-hydroxyaldehydes have been previously identified as products of lipid peroxidation, and although they represent the simplest reducing sugars, their potential for modification of proteins under physiological conditions has not been investigated. Here, 2-hydroxyaldehydes were found to condense with amines in two ways, implicating potential pathways for modification of lysine residues. A fluorescent 4,5-dialkyl-3-hydroxypyridinium with ex/em 327/390 nm and a nonfluorescent 4-alkylimidazolium cross-linking product were isolated and characterized by (1)H NMR, (13)C NMR, high-resolution mass spectrometry, and, in the former case, through independent synthesis. Both reactions appear to proceed through Amadori rearrangement of the initial Schiff base. On the basis of the UV absorbance of the 3-hydroxypyridinium, the latter was estimated to represent modification of 1.5% of the lysines of RNase incubated with 0.5 mM 2 hydroxyheptanal for 10 days at 25 degrees C. The 4-alkylimidazolium is proposed to contribute to the protein cross-linking observed by gel electrophoresis in the incubation of RNase with higher concentrations of 2-hydroxyheptanal. PMID- 12588196 TI - Mechanism of nickel assault on the zinc finger of DNA repair protein XPA. AB - Xeroderma pigmentosum group A complementing protein (XPA) is a member of the protein complex of the nucleotide excision repair (NER) pathway of DNA repair, participating in the assembly of the incision complex. The 4S zinc finger domain of XPA is involved the interactions with other NER proteins. As demonstrated previously, the activity of XPA is compromised by several metal ions implicated in DNA repair inhibition, including Ni(II), Cd(II), and Co(II) (Asmuss, M., Mullenders, L. H. F., Elker, A., and Hartwig, A. (2000) Carcinogenesis 21, 2097 2104). To study the possible molecular mechanisms of XPA inhibition, we investigated Zn(II) and Ni(II) interactions with the synthetic 37 peptide (XPAzf), representing the XPA zinc finger sequence AcDYVICEECGKEFMDSYLMNHFDLPTCDNCRDADDKHKam. The binding constants were determined using fluorescence and UV-vis spectroscopies, structural insights were provided by CD, and oxidative damage to XPAzf was studied with HPLC. The binding constants for Zn(II) and Ni(II) are (8.5 +/- 1.5) x 10(8) (log value 8.93(7)) and (1.05 +/- 0.07) x 10(6) M(-)(1) (6.02(3)), respectively, in 10 mM phosphate buffer, pH 7.4, and (6 +/- 4) x 10(9) (9.8(2)) and (2.9 +/- 0.5) x 10(6) M(-)(1) (6.46(8)) in 50 mM phosphate buffer, pH 7.4, yielding binding constant ratios Zn(II)/Ni(II) of 800 +/- 100 and 2300 +/- 500, respectively. The Ni(II) ion forms a square planar complex with the sulfurs of XPAzf, opposed to the tetrahedral structure of the native Zn(II) complex. Consequently, the overall zinc finger structure is lost in the Ni(II)-substituted peptide. Zn(II)-saturated XPAzf is remarkably resistant to air oxidation and is only slowly oxidized by 0.01 mM, 0.1 mM, and 1 mM H(2)O(2) in a concentration-dependent fashion. However, the presence of just 10-fold molar excess of Ni(II) is sufficient to accelerate this process for all three H(2)O(2) concentrations tested. Overall, our results indicate that XPAzf can undergo Ni(II) assault in specific conditions. PMID- 12588197 TI - What to do about the metabolic syndrome? PMID- 12588198 TI - Trimethoprim-sulfamethoxazole revisited. AB - During the past 3 decades, the combination of trimethoprim and sulfamethoxazole has occupied a central role in the treatment of various commonly encountered infections and has also been particularly useful for several specific clinical conditions. However, changing resistance patterns and the introduction of newer broad-spectrum antibiotics have led to the need to carefully redefine the appropriate use of this agent in clinical practice. While trimethoprim sulfamethoxazole's traditional role as empirical therapy for several infections has been modified by increasing resistance, it remains a highly useful alternative to the new generation of expanded-spectrum agents if resistance patterns and other clinical variables are carefully considered. It also seems to have an increasing role as a cost-effective pathogen-directed therapy with the potential to decrease or delay development of resistance to newer antibiotics used for empirical treatment. In addition, trimethoprim-sulfamethoxazole continues to be the drug of choice for several clinical indications. PMID- 12588199 TI - Detection of proximal adenomatous polyps with screening sigmoidoscopy: a systematic review and meta-analysis of screening colonoscopy. AB - BACKGROUND: The relative effectiveness of flexible sigmoidoscopy compared with colonoscopy to screen for colorectal cancer depends on the magnitude of the association between findings in the proximal and distal colon and the false negative rate of screening sigmoidoscopy for proximal neoplasia. To address this, we performed a systematic review and meta-analysis of screening colonoscopy studies. METHODS: Published studies through July 31, 2000, of asymptomatic patients undergoing screening colonoscopy were identified from the MEDLINE database. We generated pooled estimates of the odds ratio for the association between findings in the distal and proximal colon and the prevalence of isolated proximal adenomatous neoplasia. RESULTS: Using the sigmoid-descending colon junction to identify the beginning of the distal colon, the pooled odds ratio for the association between distal adenomatous polyps and any proximal neoplasia was 2.40 (95% confidence interval [CI], 1.42-4.05). Diminutive distal adenomatous polyps were also associated with proximal neoplasia (odds ratio, 2.36; 95% CI, 1.30-4.29). Distal hyperplastic polyps were not associated with proximal neoplasia (odds ratio, 1.44; 95% CI, 0.79-2.62). The prevalence of isolated advanced proximal neoplasia in the 3 studies was 2%, 3%, and 5%. Using the sigmoid-descending colon junction to identify the beginning of the distal colon yields a pooled estimate of isolated proximal neoplasia of 16.3% (95% CI, 13.6% 19.1%). CONCLUSIONS: Distal adenomatous polyps, including diminutive distal adenomatous polyps, are associated with an increased prevalence of synchronous proximal neoplasia. Two percent to 5% of patients undergoing screening colonoscopy may have isolated advanced proximal neoplasia. Even more patients may have isolated nonadvanced proximal neoplasia. PMID- 12588200 TI - Colon polyp recurrence in a managed care population. AB - BACKGROUND: Detection, treatment, and subsequent screening of patients with colorectal polyps have considerable implications for public health. Understanding the natural history of colorectal polyps from biologic and health care services standpoints is critical in determining optimal treatment. METHODS: Using computerized data from a large Midwestern health maintenance organization, we identified patients 50 years or older who underwent a polypectomy (index polypectomy) between January 1, 1989, and December 31, 1999. We followed up patients to identify subsequent polypectomies through September 1, 2001. RESULTS: Our final study sample consisted of 8865 individuals with an index polypectomy. Overall, 2704 patients (30.5%) were diagnosed as having recurrent polyps. Kaplan Meier projections estimate that 50% of patients will have a recurrence within 7.6 years (95% confidence interval [CI], 7.2-8.1 years). Among patients who underwent colon screening at least 9 months after the index polypectomy (52% of original cohort), Kaplan-Meier projections estimate that 50% will have a recurrent polyp within 3.9 years (95% CI, 3.8-4.1 years). CONCLUSIONS: The results demonstrate that, even when screening and treatment are received by those who need it, the risk of colon polyp recurrence is high and many patients do not undergo additional screening. Efforts to increase and monitor ongoing screening of postpolypectomy patients are warranted. In addition, further research is needed to better identify patients at risk for recurrence so that continued screening and other interventions can be targeted at these groups. PMID- 12588201 TI - The metabolic syndrome: prevalence and associated risk factor findings in the US population from the Third National Health and Nutrition Examination Survey, 1988 1994. AB - BACKGROUND: The metabolic syndrome is an important cluster of coronary heart disease risk factors with common insulin resistance. The extent to which the metabolic syndrome is associated with demographic and potentially modifiable lifestyle factors in the US population is unknown. METHODS: Metabolic syndrome associated factors and prevalence, as defined by Adult Treatment Panel III criteria, were evaluated in a representative US sample of 3305 black, 3477 Mexican American, and 5581 white men and nonpregnant or lactating women aged 20 years and older who participated in the cross-sectional Third National Health and Nutrition Examination Survey. RESULTS: The metabolic syndrome was present in 22.8% and 22.6% of US men and women, respectively (P =.86). The age-specific prevalence was highest in Mexican Americans and lowest in blacks of both sexes. Ethnic differences persisted even after adjusting for age, body mass index, and socioeconomic status. The metabolic syndrome was present in 4.6%, 22.4%, and 59.6% of normal-weight, overweight, and obese men, respectively, and a similar distribution was observed in women. Older age, postmenopausal status, Mexican American ethnicity, higher body mass index, current smoking, low household income, high carbohydrate intake, no alcohol consumption, and physical inactivity were associated with increased odds of the metabolic syndrome. CONCLUSIONS: The metabolic syndrome is present in more than 20% of the US adult population; varies substantially by ethnicity even after adjusting for body mass index, age, socioeconomic status, and other predictor variables; and is associated with several potentially modifiable lifestyle factors. Identification and clinical management of this high-risk group is an important aspect of coronary heart disease prevention. PMID- 12588202 TI - Physician, public, and policymaker perspectives on chronic conditions. AB - BACKGROUND: Chronic conditions are the leading reason why people seek medical care, yet the current financing and delivery system has been criticized for not promoting ongoing care. The perceptions of physicians, policymakers, and the general public were compared on how well the current system addresses the needs of people with chronic conditions. METHODS: National surveys of 1238 physicians and 1663 Americans and a convenience sample of 155 policymakers were compared. All 3 groups were given the same definition of a chronic condition and asked similar questions. RESULTS: There was strong agreement that chronic medical conditions affect men and women of all ages, ethnicities, and income levels (>90% strongly or somewhat agree). However, compared with the public and physicians, policymakers were less likely to respond that people with chronic conditions usually receive adequate medical care, that health insurance pays for most needed services, or that government programs are adequate. The public was most positive about the current system and policymakers the least. A majority of all 3 groups agreed that it is somewhat or very difficult for people with chronic conditions to obtain adequate care from primary care physicians, medical specialists, and other health care professionals. CONCLUSIONS: A majority of physicians, policymakers, and the general public are concerned that the current health care system is not addressing the needs of people with chronic conditions. Changes in how medical care is financed and delivered are necessary to respond to these concerns. PMID- 12588203 TI - Trends in the mortality burden associated with diabetes mellitus: a population based study in Rochester, Minn, 1970-1994. AB - BACKGROUND: The prevalence of diabetes mellitus (DM) has increased markedly in recent decades, but trends in the mortality burden associated with DM are unclear. Therefore, we analyzed population-based longitudinal data to address this issue. METHODS: The community-based medical records of all Rochester residents 45 years and older who died between January 1, 1970, and December 31, 1994, were reviewed to identify those who met the standardized criteria for DM before death. Trends over successive quinquenniums were assessed for the proportion of all deaths in the community of persons with prevalent DM, for mortality rates for persons with and without DM, and for the distribution of causes of death among decedents with and without DM. RESULTS: Of 10 152 total deaths in 1970-1994, 1384 (13.6%) met the criteria for prevalent DM. Between 1970 1974 and 1990-1994, the proportion of decedents with DM increased by 48.2%. Mortality rates for persons with and without DM declined by 13.8% and 21.4%, respectively. This disparity in mortality trends was most apparent for older women and younger men. There were temporal declines in the proportion of all persons dying of cardiovascular disease, but temporal declines in persons dying of cerebrovascular disease were found only in decedents without DM. CONCLUSIONS: The mortality burden associated with DM increased significantly between 1970 and 1994, probably due to increases in DM incidence and smaller declines in mortality for persons with DM relative to those without DM. In the absence of improved DM prevention and treatment, the steady declines in mortality observed for the general population since the 1960s will likely begin to slow or even reverse. PMID- 12588204 TI - Imaging characteristics of a novel technetium Tc 99m-labeled platelet glycoprotein IIb/IIIa receptor antagonist in patients With acute deep vein thrombosis or a history of deep vein thrombosis. AB - BACKGROUND: The diagnosis of recurrent deep vein thrombosis (DVT) is challenging. Imaging with radiolabeled peptides offers a new approach for detecting acute DVT. Technetium Tc 99m ((99m)Tc)-apcitide binds with high affinity and specificity to the glycoprotein IIb/IIIa receptors expressed on activated platelets and, therefore, (99m)Tc-apcitide scintigraphy should be negative with residual abnormalities caused by old, inactive thrombi and positive with new, active thrombi. METHODS: In a prospective multicenter study, (99m)Tc-apcitide imaging was performed on 38 patients with a newly diagnosed first DVT (group 1) and 40 patients with previous DVT, symptoms of postthrombotic syndrome, and chronic intraluminal abnormalities on ultrasonography (group 2). Images were interpreted in a blinded fashion by 2 experts and by newly trained nuclear medicine physicians. The sensitivity and specificity of (99m)Tc-apcitide were determined by calculating the proportion of scans in group 1 patients that were read as "positive for acute DVT" and the proportion of scans in group 2 patients that were read as "negative for acute DVT," respectively. RESULTS: When read by 2 experts, ( 99m)Tc-apcitide had a sensitivity of 92% for both readers and specificities of 82% and 90%. Agreement between the experts was excellent. However, the accuracy and interreader agreement for newly trained nuclear medicine physicians were lower. CONCLUSIONS: Technetium Tc 99m-apcitide scintigraphy has potential utility in suspected recurrent DVT because it detects most acute thrombi and has few false-positive results in patients with previous DVT. However, the accuracy appears to depend on the training and experience of the interpreters. PMID- 12588205 TI - Acute myocardial infarction in human immunodeficiency virus-infected patients. AB - BACKGROUND: Patients infected with human immunodeficiency virus (HIV) are at an increased risk for premature coronary artery disease. However, the clinical outcome of HIV-infected patients who have had an acute myocardial infarction (AMI) is unknown. METHODS: We studied 24 consecutive HIV-infected patients admitted because of AMI. During the hospital phase, the patients were examined for recurrent ischemia, congestive heart failure, arrhythmia, and death. Patients were followed up for an average of 15 months after discharge for reinfarction; recurrent angina; the need for any angioplasty, bypass surgery, or target vessel revascularization for restenosis and stent thrombosis; HIV-related complications; and death. For comparison, we included a matched control group of non-HIV infected patients. RESULTS: The HIV-infected patients with AMI were predominantly male (21 [88%]), 47 +/- 9 years of age. Twenty-two (92%) were receiving antiretroviral treatment; 17 (71%), protease inhibitors; and 13 (54%), lipid lowering therapy. With aggressive therapy, the lipid profile was similar in HIV infected patients treated with protease inhibitors and those who were not. Twenty one (88%) of 24 patients underwent immediate angiography and 20 (83%) had angioplasty or bypass surgery. The HIV-infected patients with AMI had a benign in hospital course, with no deaths or reinfarction. The admission characteristics, treatment strategy, and in-hospital outcome were similar in the matched uninfected patients with AMI. After discharge, HIV-infected patients had a higher incidence of reinfarction (4/20 [20%] vs 2/45 [4%]; P =.07), and 6 (43%) of 14 HIV-infected patients who had successful percutaneous coronary intervention and were available for follow-up required target vessel revascularization compared with 4 (11%) of 38 uninfected patients who had successful percutaneous coronary intervention and were available for follow-up (P =.02). CONCLUSIONS: Patients infected with HIV sustain AMI at a young age and have a benign in-hospital course. Although HIV-infected patients have a higher incidence of postdischarge ischemic events, restenosis, and stent thrombosis, the intermediate-term mortality is low. PMID- 12588206 TI - Differential use of coronary revascularization and hospital mortality following acute myocardial infarction. AB - BACKGROUND: The effect of racial/ethnic disparity in the use of cardiac procedures on short-term outcomes, such as hospital mortality, is limited. We sought to determine the association of revascularization procedures (percutaneous transluminal coronary angioplasty or coronary artery bypass graft) to hospital mortality in non-Hispanic black and white patients and Hispanic patients with acute myocardial infarction. METHODS: Analysis of the New York State Department of Health Statewide Planning and Research Cooperate System (SPARCS) data for 12 555 patients admitted to New York City hospitals with acute myocardial infarction in 1996. Revascularization procedure frequencies and adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for hospital mortality were calculated. RESULTS: Whites were older than Hispanics and blacks (mean +/- SD age, 70 +/- 13.3 vs 64 +/- 13.3 and 64 +/- 12.9 years, respectively; P<.001) and more likely to have heart failure (36.3% vs 29.1% and 29.6%, respectively; P<.001). Blacks were least likely to be revascularized compared with Hispanics and whites (15.8% vs 25.8% and 25.2%, respectively; P<.001). Hispanics were more likely to survive than whites (adjusted OR, 0.73 [95% CI, 0.59-0.91]); this difference was not significant for blacks (adjusted OR, 0.83 [95% CI, 0.69-1.00]). Nonrevascularized blacks and Hispanics were more likely to be discharged alive than nonrevascularized whites (OR, 0.80 [95% CI, 0.66-0.98] for blacks; OR, 0.74 [95% CI, 0.59-0.93] for Hispanics). There were no significant racial/ethnic differences in hospital survival among revascularized patients. CONCLUSIONS: Nonclinical and clinical factors appear to account for blacks being least likely to have been revascularized. Despite these differences in revascularization rates, survival was similar for blacks and whites, whereas Hispanics were more likely to survive than whites. PMID- 12588207 TI - Clinical and epidemiologic features of group a streptococcal pneumonia in Ontario, Canada. AB - BACKGROUND: Since the 1960s, group A streptococcus (GAS) has accounted for less than 1% of cases of community-acquired pneumonia. During the past 2 decades there has been a resurgence of invasive GAS infection, but no large study of GAS pneumonia has been performed. METHODS: To determine the clinical and epidemiologic features of GAS pneumonia, we conducted prospective, population based surveillance of all invasive GAS infection in residents of Ontario from January 1, 1992, through December 31, 1999. RESULTS: Of 2079 cases of invasive GAS infection, 222 (11%) represented GAS pneumonia. The incidence of GAS pneumonia ranged from 0.16 per 100 000 in 1992 to 0.35 per 100 000 in 1999. Most cases were community acquired (81%). Forty-four percent of nursing home-acquired cases occurred during outbreaks. The case fatality rate was 38% for GAS pneumonia, compared with 12% for the entire cohort with invasive GAS infection and 26% for patients with necrotizing fasciitis. The presence of streptococcal toxic shock syndrome (odds ratio, 19; 95% confidence interval, 8.4-42; P =.001) and increasing age (odds ratio per decade, 1.45; 95% confidence interval, 1.2 1.7; P<.001) were associated with fatal outcome. Time to death was rapid, with a median of 2 days despite antimicrobial therapy and supportive measures. CONCLUSIONS: Group A streptococcal pneumonia is a common form of invasive GAS disease but remains an uncommon cause of community-acquired pneumonia. Progression is rapid despite appropriate therapy. The incidence is similar to, and the case fatality rate higher than, that of necrotizing fasciitis. PMID- 12588208 TI - Role of organizational factors in poor blood pressure control in patients with type 2 diabetes: the QuED Study Group--quality of care and outcomes in type 2 diabetes. AB - BACKGROUND: A large body of evidence supports the need for reducing the cardiovascular burden of diabetes. Only indirect and occasional data describe the adequacy of routine management of hypertension in patients with diabetes. The aim of this study was to explore the interplay of some potential key determinants of quality of antihypertensive care, including the settings, physicians' beliefs about blood pressure (BP) control, and patient-related factors. METHODS: We evaluated physicians' beliefs about BP control using questionnaire responses at study entry. A sample of 3449 patients with type 2 diabetes mellitus, of whom 1782 (52%) were considered to have hypertension, was recruited by 212 physicians practicing in 125 diabetes outpatients clinics (DOCs) and 106 general practitioners (GPs). We evaluated the type and number of antihypertensive agents used and the BP values at study entry and after 1 year of follow-up. We used multilevel analysis to investigate correlates of poor BP control (> or =160/90 mm Hg). RESULTS: Only 16% of GPs and 14% of DOC physicians targeted BP values of less than 130/85 mm Hg. At study entry, 6% of the patients had values below 130/85 mm Hg, whereas 52% showed values of 160/90 mm Hg or greater. Only 12% of subjects were treated with more than 2 drugs at study entry, compared with 16% at the 1-year follow-up (P =.001). Multilevel analysis showed that patients attending DOCs had a more than 2-fold increased risk for inadequate BP control, compared with those treated by GPs. The risk for poor BP control was 2 times higher for patients treated by male physicians compared with those treated by female physicians, and it was halved when the physician responsible for the diabetes care specialized in diabetology or endocrinology. CONCLUSION: In a model situation of comorbidity, the overall quality of care depends on structural and organizational factors, which are likely to be more influential than existing guidelines. PMID- 12588209 TI - Effect of selective cyclooxygenase 2 inhibitors and naproxen on short-term risk of acute myocardial infarction in the elderly. AB - BACKGROUND: Recent debate has emerged regarding the cardiovascular safety of selective cyclooxygenase 2 inhibitors and the possible cardioprotective effect of naproxen sodium. We compared the rates of acute myocardial infarction (AMI) among elderly patients dispensed selective cyclooxygenase 2 inhibitors, naproxen, and nonselective nonnaproxen nonsteroidal anti-inflammatory drugs (NSAIDs). METHODS: We conducted a population-based retrospective cohort study using administrative health care data from Ontario, Canada, from April 1, 1998, to March 31, 2001. We identified NSAID-naive cohorts of subjects aged 66 years and older in whom treatment was initiated with celecoxib (n = 15 271), rofecoxib (n = 12 156), naproxen (n = 5669), and nonnaproxen nonselective NSAIDs (n = 33 868), along with a randomly selected control cohort not exposed to NSAIDs (n = 100 000). Multivariate Cox proportional hazards models were used to compare AMI rates between study drug groups while controlling for potential confounders. RESULTS: Relative to control subjects, the multivariate model showed no significant differences in AMI risk for new users of celecoxib (adjusted rate ratio [aRR], 0.9; 95% confidence interval [CI], 0.7-1.2), rofecoxib (aRR, 1.0; 95% CI, 0.8 1.4), naproxen (aRR, 1.0; 95% CI, 0.6-1.7), or nonnaproxen nonselective NSAIDs (aRR, 1.2; 95% CI, 0.9-1.4). CONCLUSIONS: The findings of this observational study suggest no increase in the short-term risk of AMI among users of selective cyclooxygenase 2 inhibitors as commonly used in clinical practice. Furthermore, the findings do not support a short-term reduced risk of AMI with naproxen. PMID- 12588210 TI - The economic burden of non-influenza-related viral respiratory tract infection in the United States. AB - BACKGROUND: Viral respiratory tract infection (VRTI) is the most common illness in humans. Despite the high incidence, the economic impact of non-influenza related VRTI has not been rigorously explored. Our objectives were to obtain an updated incidence of non-influenza-related VRTI in the United States and to quantify the health care resource use (direct costs) and productivity losses (indirect costs) associated with these infections. METHODS: A nationwide telephone survey of US households (N = 4051) was conducted between November 3, 2000, and February 12, 2001 to obtain a representative estimate of the self reported incidence of non-influenza-related VRTI and related treatment patterns. Direct treatment costs measured included outpatient clinician encounters, use of over-the-counter and prescription drugs, and associated infectious complications of non-influenza-related VRTI. Absenteeism estimates for infected individuals and parents of infected children were extrapolated from National Health Interview Survey data. RESULTS: Of survey respondents, 72% reported a non-influenza-related VRTI within the past year. Respondents who experienced a self-reported non influenza-related VRTI averaged 2.5 episodes annually. When these rates are extrapolated to the entire US population, approximately 500 million non-influenza related VRTI episodes occur per year. Similarly, if the treatment patterns reported by the respondents are extended to the population, the total economic impact of non-influenza-related VRTI approaches $40 billion annually (direct costs, $17 billion per year; and indirect costs, $22.5 billion per year). CONCLUSIONS: Largely because of the high attack rate, non-influenza-related VRTI imposes a greater economic burden than many other clinical conditions. The pending availability of effective antiviral therapies warrants increased attention be paid to this common and expensive illness. PMID- 12588211 TI - Risk of hepatitis C virus transmission from infected sanitary staff to patients. PMID- 12588213 TI - Medical treatment of acute illnesses in end-stage dementia. PMID- 12588215 TI - Fondaparinux requires further study before firm recommendation. PMID- 12588216 TI - Depression and negative outcomes in patients with heart failure. PMID- 12588218 TI - Successful treatment of granulomatous cheilitis with thalidomide. PMID- 12588219 TI - Fluoroscopy-induced skin necrosis. PMID- 12588220 TI - A prospective evaluation of the incidence of complications associated with Mohs micrographic surgery. AB - BACKGROUND: Because outpatient surgery is being increasingly scrutinized in the lay press, it is important that dermatologists and dermatologic surgeons accurately characterize the safety of office-based surgery. Although there is abundant anecdotal evidence to support the inherent safety of dermatologic surgery, there are few data that support the safety of Mohs micrographic surgery (MMS) as performed by appropriately trained dermatologic surgeons in outpatient settings. DESIGN: All patients presenting for MMS micrographic surgery during the calendar year 2000 were prospectively enrolled in this study designed to evaluate the incidence of multiple complications associated with scalpel-based cutaneous surgery (postoperative hemorrhage, hematoma formation, wound infection, wound dehiscence, and flap/graft necrosis). SETTING: An academic MMS practice. PATIENTS: A total of ,1052 patients (1,358 MMS cases) were prospectively enrolled. Complete follow-up information was available for 1,343 cases (98.9%). RESULTS: Complications associated with MMS were very infrequent, with an overall complication incidence of 1.64% (22/1,343). Most surgical complications involved difficulties with hemostasis. No complications were significant enough to involve the assistance of another specialist or to require the hospitalization of the patient. CONCLUSIONS: Mohs micrographic surgery is a very safe outpatient procedure when performed by appropriately trained physicians. The types of complications seen in our patients were identical to those seen in hospitalized patients described in previous studies. Our complication rates were equal to or lower than the published complication rates from specialists in other surgical disciplines. PMID- 12588221 TI - Narrowband (312-nm) UV-B suppresses interferon gamma and interleukin (IL) 12 and increases IL-4 transcripts: differential regulation of cytokines at the single cell level. AB - OBJECTIVE: To determine whether 312-nm UV-B alters production of effector and regulatory cytokines by viable T cells that remain in psoriatic lesions during UV B phototherapy. DESIGN: Prospective study. SETTING: General clinical research center of The Rockefeller University Hospital. PATIENTS: Ten adult patients with moderate to severe psoriasis vulgaris that was difficult to manage were sequentially enrolled in our protocols, and biopsies were taken at various time points from resolving lesions. INTERVENTION: Narrowband (312-nm) UV-B was given starting at 50% of a minimum erythema dose, then increased daily 10% to 15% if no apparent erythema was induced. Patients continued with treatment until maximal benefit was noted. In some experiments, T cells were irradiated ex vivo with standard TL-01 fluorescent bulbs (Philips Lighting Co, Somerset, NJ). MAIN OUTCOME MEASURES: Intracellular cytokine staining was done using flow cytometry to quantify numbers of cytokine-producing cells from epidermal and peripheral T cells. The production of messenger RNA for interleukin (IL) 12, interferon (IFN) gamma, tumor necrosis factor alpha, IL-4, and IL-10 was measured by quantitative reverse transcription-polymerase chain reaction. RESULTS: Ultraviolet-B treatment eliminated production of IL-12 messenger RNA and decreased production of IFN gamma messenger RNA by more than 60% in irradiated psoriasis lesions (P<.03 for both). Within 1 to 2 weeks of starting UV-B treatment, the frequency of viable T cells producing IFN-gamma decreased 40% to 65%. In contrast, mRNA for IL-4 increased by 82% (P =.05) during UV-B treatment, and the number of IL-4-producing cells increased by 228% after 1 week of treatment. In vitro experiments established that, on the single-cell level, survival and cytokine production by type 1 T cells were differentially regulated by UV-B. CONCLUSIONS: Therapeutic UV B suppresses the type 1 (proinflammatory) axis as defined by IL-12, IFN-gamma, and IL-8, and can selectively reduce proinflammatory cytokine production by individual T cells. Knowledge of the immunomodulatory effects of UV-B will help to integrate this modality in future therapeutics for psoriasis based on deliberate blockade of inflammatory molecular pathways in the type 1 T-cell pathway. PMID- 12588222 TI - Topical nitrogen mustard in the management of mycosis fungoides: update of the Stanford experience. AB - OBJECTIVE: To evaluate and update the response and survival outcomes and toxic effects in patients treated with topical nitrogen mustard (mechlorethamine hydrochloride) as primary therapy. DESIGN: A single-center, retrospective cohort analysis. SETTING: Academic referral center for cutaneous lymphoma. PATIENTS: A total of 203 patients with mycosis fungoides (clinical stages I-III) treated with topical nitrogen mustard as initial therapy. MAIN OUTCOME MEASURES: Long-term actuarial survival, freedom-from-relapse, and freedom-from-progression results as calculated by the Kaplan-Meier method. RESULTS: The overall response rate for the 203 patients was 83%, with a complete response rate of 50%. The median time to achieve complete response was 12 months (T1, 10 months; T2, 19 months), and the median time to relapse was 12 months. The duration of complete response increased with longer maintenance therapy; however, after completion of therapy, the response duration or relapse rate was similar regardless of maintenance regimen. Patients with T1 disease had better response and survival outcomes than those with T2 disease, with overall and complete response rates in T1 of 93% and 65%, respectively, and in T2, 72% and 34%, respectively. A similar clinical response was seen for patients with stage IIA vs IB. Sixty-eight percent of 203 patients received only topical nitrogen mustard therapy throughout their follow-up course, including most of the patients who achieved an initial complete response. The clinical response to topical nitrogen mustard as salvage therapy was similar to initial response rates. The efficacy results were similar in patients treated with aqueous vs ointment preparations. Freedom-from-progression rates in T1 disease (no progression to higher T classification or worse clinical stage) at 5 and 10 years were 92% and 85%, respectively, and in T2, 83% at 5 and 10 years. Fewer than 10% of patients experienced contact hypersensitivity reactions when topical nitrogen mustard was used as an ointment preparation. Only 8 patients (4%) developed secondary cutaneous malignancy, none attributable to topical nitrogen mustard monotherapy. Pediatric patients experienced no significant toxic effects with topical nitrogen mustard therapy. CONCLUSIONS: Topical nitrogen mustard remains an effective primary initial or salvage therapy in mycosis fungoides for patients with T1 and T2 disease. Long-term follow-up results confirm its safety. PMID- 12588223 TI - 9-cis-retinoic acid capsules in the treatment of AIDS-related Kaposi sarcoma: results of a phase 2 multicenter clinical trial. AB - OBJECTIVE: To evaluate the safety, dose tolerance, and anti-tumor effects of 9 cis-retinoic acid in the treatment of Kaposi sarcoma (KS) related to acquired immunodeficiency syndrome (AIDS). DESIGN: Phase 2, open-label clinical trial of oral doses of 9-cis-retinoic acid increasing in 40-mg increments every 2 weeks from 60 mg/m(2) per day to a maximum of 140 mg/m( 2) per day. SETTING: Five hospital or health maintenance organization outpatient clinics. PATIENTS: Fifty seven adult male patients with human immunodeficiency virus and biopsy-proven KS. MAIN OUTCOMES MEASURES: Safety was evaluated by adverse events, physical examination, laboratory test abnormalities, treatment-limiting toxic effects, and reasons for early withdrawal. Response (>/=50% improvement) was evaluated by an overall KS response and by the area and height from 6 index lesions selected at baseline. RESULTS: Patients tolerated 60 and 100 mg/m(2) per day. Most patients found 140 mg/m(2) per day intolerable owing to headache. Common treatment-related adverse events were headache, xerosis, rash, alopecia, and hyperlipemia. The patient response rate for the overall KS disease was 19% (11/57), including 1 patient with clinically complete response. The response rate assessed by measuring 6 index lesions during treatment was 39% (22/57). Sixteen responding patients (73%) were refractory to at least 1 previous anti-KS therapy. Patients with CD4( +) counts of 150 cells/ micro L or lower were as likely to respond as patients with counts of higher than 150 cells/ micro L. The median time to response was 8.5 weeks (range, 4.0-21.1 weeks). The median duration of treatment was 15.1 weeks (range, 0.14 to >/=62 weeks). CONCLUSION: 9-cis-retinoic acid capsules have moderate activity and provide durable responses, but substantial toxic effects at higher doses limit its suitability as an anti-KS therapy. PMID- 12588225 TI - Improved identification of potentially dangerous pigmented skin lesions by computerized image analysis. AB - BACKGROUND: Melanoma is completely curable if resected early. Unfortunately, early melanoma can be difficult to differentiate from other pigmented lesions. Computerized image analysis instruments have now been developed to assist in determining whether a pigmented lesion is potentially dangerous and requires biopsy. To evaluate whether one such instrument can improve the management of pigmented lesions, we obtained biopsy specimens from 52 pigmented lesions that appeared clinically benign to an experienced dermatologist but were suspicious by image analysis. OBSERVATION: Histologically, 9 (17%) of the lesions that were removed based solely on computer recommendation were potentially dangerous and should have been removed. These included 1 malignant melanoma in situ and 8 dysplastic nevi with moderate to severe cytologic atypia. CONCLUSION: The results of the present study indicate that computerized image analysis can improve the evaluation of pigmented skin lesions by identifying clinically unsuspicious, but potentially dangerous, lesions that might have otherwise have been neglected. PMID- 12588224 TI - Clinical risk factors for mortality in patients with neurofibromatosis 1: a cohort study of 378 patients. AB - OBJECTIVE: To identify the main clinical features associated with mortality in patients with neurofibromatosis 1. DESIGN: Cohort study. SETTING: Referral center for neurofibromatosis. PATIENTS: Three hundred seventy-eight patients with neurofibromatosis 1 who had more than 1 year of follow-up in the center. MAIN OUTCOME MEASURES: Mortality. Clinical features, especially dermatological, were evaluated as potential factors associated with mortality. RESULTS: Factors associated independently with mortality were the presence of subcutaneous neurofibromas (odds ratio, 10.8; 95% confidence interval, 2.1-56.7; P<.001), the absence of cutaneous neurofibromas (odds ratio, 5.3; 95% confidence interval, 1.2 25.0; P =.03), and facial asymmetry (odds ratio, 11.4; 95% confidence interval, 2.6-50.2; P<.01). CONCLUSIONS: Some features that can be found by a routine clinical examination are associated with mortality in patients with neurofibromatosis 1. Clinical follow-up should be focused on patients with subcutaneous neurofibromas and/or the absence of cutaneous neurofibromas and/or facial asymmetry. PMID- 12588226 TI - Late recurrence of inflammatory first-stage lesions in incontinentia pigmenti: an unusual phenomenon and a fascinating pathologic mechanism. AB - BACKGROUND: Incontinentia pigmenti (IP) is an X-linked genodermatosis that is manifested by neonatal inflammatory vesicles localized along the lines of Blaschko. These lesions usually clear spontaneously within a few months, leaving hyperpigmentation. Ophthalmologic and neurologic symptoms can be associated with IP. Late recurrences of the first-stage inflammatory lesions after the initial rash are uncommon and have been reported infrequently. The mechanism involved in this phenomenon is unclear. However, the recent identification of NEMO/IKKgamma as the gene responsible for IP sheds new light on its pathophysiologic origins. OBSERVATIONS: We report 5 cases of children who experienced episodes of late reactivation of IP. In all cases, the recurrences occurred on the previously hyperpigmented streaks several months or years after resolution of the initial eruptions. In most cases, the recurrences were preceded by an infectious episode. CONCLUSIONS: These IP recurrences suggest that mutated cells can persist a long time in the epidermis. We theorize that infections trigger the reactivations. The NEMO/IKKgamma gene encodes a protein essential in nuclear factor kappaB activation, which is required for resistance to tumor necrosis factor alpha induced apoptosis. We discuss the role of a proinflammatory cytokine such as tumor necrosis factor alpha as a triggering factor for the reactivation. PMID- 12588227 TI - Congenital dermatofibrosarcoma protuberans: variability in presentation. AB - BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is an uncommon low-grade fibrohistiocytic tumor that usually occurs on the trunk or proximal extremities and typically appears during the second to fifth decade of life. It most commonly begins as a red-blue plaque that grows slowly and ultimately becomes nodular. The tumor is associated with a high recurrence rate but low metastatic potential. It rarely presents in childhood and is even more rarely present at birth. The clinical diagnosis of DFSP in infancy or childhood may be difficult because, in its early stages, the tumor often resembles a vascular birthmark. OBSERVATIONS: We studied 6 patients with congenital DFSP who were initially thought to have other diagnoses, highlighting the potential clinical variability in presentation. Half of the cases in this series occurred in areas of the body outside of the typically reported distribution pattern of acquired DFSP and in locations that, therefore, may not arouse suspicion of congenital DFSP. CONCLUSIONS: Given the aggressive local potential and high recurrence rate of DFSP, early diagnosis is preferable to facilitate appropriate excision. We recommend that any infant or child presenting with a cutaneous plaque or nodule, even congenital, that does not have characteristic or diagnostic clinical features undergo tissue biopsy for histologic evaluation. PMID- 12588228 TI - Complications in dermatologic surgery: how safe is safe? PMID- 12588229 TI - Violaceous plaques in a patient with acquired immunodeficiency syndrome. PMID- 12588230 TI - Multiple bilateral vesiculobullous lesions on the lower extremities. PMID- 12588231 TI - Multiple facial papules in an adult. PMID- 12588232 TI - Bilateral periorbital eruption in an immunocompromised host. PMID- 12588233 TI - Narrowband UV-B vs medium-dose UV-A1 phototherapy in chronic atopic dermatitis. PMID- 12588234 TI - The acute edema/cutaneous distension syndrome. PMID- 12588236 TI - Topical corticosteroids and bullous pemphigoid. PMID- 12588237 TI - Systemic adverse effects after botulinum toxin type B (myobloc) injections for the treatment of palmar hyperhidrosis. PMID- 12588238 TI - Pseudoporphyria induced by oral contraceptive pills. PMID- 12588239 TI - Direct immunofluorescence of plucked hair in pemphigus. PMID- 12588240 TI - A rosacea-like granulomatous eruption in a patient using tacrolimus ointment for atopic dermatitis. PMID- 12588245 TI - A piece of my mind. An essay on desire. PMID- 12588247 TI - Web site showcases science-based information on herbs, other supplements. PMID- 12588246 TI - Survey suggesting that prescription drug ads help public is met with skepticism. PMID- 12588248 TI - South Korean physician named to lead WHO. PMID- 12588254 TI - Exposure to pets in childhood and risk of atopic disorders. PMID- 12588256 TI - Exposure to pets in childhood and risk of atopic disorders. PMID- 12588257 TI - Exposure to pets in childhood and risk of atopic disorders. PMID- 12588258 TI - Exposure to pets in childhood and risk of atopic disorders. PMID- 12588259 TI - Lifestyle modifications to prevent hypertension. PMID- 12588260 TI - Lifestyle modifications to prevent hypertension. PMID- 12588263 TI - Risk of cutaneous vaccinia from health care workers who receive smallpox vaccine. PMID- 12588264 TI - Public disclosure of health plan quality of care. PMID- 12588265 TI - Public disclosure of health plan quality of care. PMID- 12588266 TI - Public disclosure of health plan quality of care. PMID- 12588267 TI - Public disclosure of health plan quality of care. PMID- 12588269 TI - Bivalirudin and provisional glycoprotein IIb/IIIa blockade compared with heparin and planned glycoprotein IIb/IIIa blockade during percutaneous coronary intervention: REPLACE-2 randomized trial. AB - CONTEXT: The direct thrombin inhibitor bivalirudin has been associated with better efficacy and less bleeding than heparin during coronary balloon angioplasty but has not been widely tested during contemporary percutaneous coronary intervention (PCI). OBJECTIVE: To determine the efficacy of bivalirudin, with glycoprotein IIb/IIIa (Gp IIb/IIIa) inhibition on a provisional basis for complications during PCI, compared with heparin plus planned Gp IIb/IIIa blockade with regard to protection from periprocedural ischemic and hemorrhagic complications. DESIGN, SETTING, AND PARTICIPANTS: The Randomized Evaluation in PCI Linking Angiomax to Reduced Clinical Events (REPLACE)-2 trial, a randomized, double-blind, active-controlled trial conducted among 6010 patients undergoing urgent or elective PCI at 233 community or referral hospitals in 9 countries from October 2001 through August 2002. INTERVENTIONS: Patients were randomly assigned to receive intravenous bivalirudin (0.75-mg/kg bolus plus 1.75 mg/kg per hour for the duration of PCI), with provisional Gp IIb/IIIa inhibition (n = 2999), or heparin (65-U/kg bolus) with planned Gp IIb/IIIa inhibition (abciximab or eptifibatide) (n = 3011). Both groups received daily aspirin and a thienopyridine for at least 30 days after PCI. MAIN OUTCOME MEASURES: The primary composite end point was 30-day incidence of death, myocardial infarction, urgent repeat revascularization, or in-hospital major bleeding; the secondary composite end point was 30-day incidence of death, myocardial infarction, or urgent repeat revascularization. RESULTS: Provisional Gp IIb/IIIa blockade was administered to 7.2% of patients in the bivalirudin group. By 30 days, the primary composite end point had occurred among 9.2% of patients in the bivalirudin group vs 10.0% of patients in the heparin-plus-Gp IIb/IIIa group (odds ratio, 0.92; 95% confidence interval, 0.77-1.09; P =.32). The secondary composite end point occurred in 7.6% of patients in the bivalirudin vs 7.1% of patients in the heparin-plus-Gp IIb/IIIa groups (odds ratio, 1.09; 95% confidence interval 0.90-1.32; P =.40). Prespecified statistical criteria for noninferiority to heparin plus Gp IIb/IIIa were satisfied for both end points. In-hospital major bleeding rates were significantly reduced by bivalirudin (2.4% vs 4.1%; P<.001). CONCLUSIONS: Bivalirudin with provisional Gp IIb/IIIa blockade is statistically not inferior to heparin plus planned Gp IIb/IIIa blockade during contemporary PCI with regard to suppression of acute ischemic end points and is associated with less bleeding. PMID- 12588270 TI - Enrollee appeals of preservice coverage denials at 2 Health Maintenance Organizations. AB - CONTEXT: Congress and state legislatures are considering patient bills of rights that seek to strengthen opportunities for patients to have denials of coverage reconsidered by their health plans. Little is publicly known about such appeals systems. OBJECTIVE: To improve understanding of the sources, types, and outcomes of conflicts between patients and managed care organizations over coverage of services. DESIGN AND SETTING: Descriptive study of information abstracted from 1774 preservice appeals out of a larger stratified random sample of 3519 appeals lodged between January 1998 and June 2000 at 2 large US health maintenance organizations. MAIN OUTCOME MEASURES: Classification of preservice appeals according to whether they contested access to out-of-network care, the contractual limits of coverage, or the medical necessity of services; analysis of contractual coverage and medical necessity appeals by the services in dispute and out-of-network appeals by enrollees' reasons for seeking care; and comparison of the proportions of appeals won by enrollees across types of appeals and services. RESULTS: Approximately one third (36.9%) of preservice appeals involved medical necessity determinations, another third (36.6%) centered on the scope of contractually covered benefits, and most of the remainder (19.7%) involved out-of network care. Enrollee wins were significantly more frequent among medical necessity appeals than out-of-network or contractual coverage appeals (52.2% vs 35.4% and 33.2%, respectively; P<.001). Appeals were concentrated among relatively few services and among therapies that are generally regarded as nonessential. CONCLUSIONS: A majority of preservice appeals disputed choice of provider or contractual coverage issues, rather than medical necessity. Medical necessity disputes proliferate not around life-saving treatments but in areas of societal uncertainty about the legitimate boundaries of insurance coverage. Greater transparency about the coverage status of specific services, through more precise contractual language and consumer education about benefits limitations, may help to avoid a large proportion of disputes in managed care. PMID- 12588271 TI - Association of serum digoxin concentration and outcomes in patients with heart failure. AB - CONTEXT: The Digitalis Investigation Group (DIG) trial reported that digoxin provided no overall mortality benefit and only a modest reduction in hospitalizations among patients with heart failure and depressed left ventricular systolic function. The clinical outcomes associated with digoxin therapy at different serum concentrations in the DIG trial have not been assessed. OBJECTIVE: To assess variations in serum digoxin concentration (SDC) and their association with mortality and hospitalization in patients with heart failure. DESIGN, SETTING, AND PATIENTS: Post hoc analysis of the randomized, double blinded, placebo-controlled DIG trial, conducted from August 1991 to December 1995, with the main analysis restricted to men with a left ventricular ejection fraction of 45% or less (n = 3782). Patients randomly assigned to receive digoxin were divided into 3 groups based on SDC at 1 month (0.5-0.8 ng/mL, n = 572; 0.9 1.1 ng/mL, n = 322; and > or =1.2 ng/mL, n = 277) and compared with patients randomly assigned to receive placebo (n = 2611). MAIN OUTCOME MEASURE: All-cause mortality at a mean follow-up of 37 months. RESULTS: Higher SDCs were associated with increased crude all-cause mortality rates (0.5-0.8 ng/mL, 29.9%; 0.9-1.1 ng/mL, 38.8%; and > or =1.2 ng/mL, 48.0%; P =.006 for trend). Patients with SDCs of 0.5 to 0.8 ng/mL had a 6.3% (95% confidence interval [CI], 2.1%-10.5%) lower mortality rate compared with patients receiving placebo. Digoxin was not associated with a reduction in mortality among patients with SDCs of 0.9 to 1.1 ng/mL (2.6% increase; 95% CI, - 3.0% to 8.3%), whereas patients with SDCs of 1.2 ng/mL and higher had an 11.8% (95% CI, 5.7%-18.0%) higher absolute mortality rate than patients receiving placebo. The association between SDC and mortality persisted after multivariable adjustment (SDC 0.5-0.8 ng/mL hazard ratio [HR] 0.80, 95% CI, 0.68-0.94; SDC 0.9-1.1 ng/mL HR 0.89, 95% CI, 0.74-1.08; SDC > or =1.2 ng/mL HR 1.16, 95% CI, 0.96-1.39; and HR of 1.00 [referent] for placebo). CONCLUSIONS: Our findings demonstrate that higher SDCs were associated with increased mortality and suggest that the effectiveness of digoxin therapy in men with heart failure and a left ventricular ejection fraction of 45% or less may be optimized in the SDC range of 0.5 to 0.8 ng/mL. PMID- 12588272 TI - Use of child booster seats in motor vehicles following a community campaign: a controlled trial. AB - CONTEXT: Once children have outgrown car seats, booster seats protect from injury better than lap and shoulder belts alone. However, the majority of children aged 4 to 8 years use only an adult seat belt. OBJECTIVE: To evaluate the effectiveness of a multifaceted community booster seat campaign in increasing observed booster seat use among child passengers in motor vehicles. DESIGN: Prospective, nonrandomized, controlled community intervention trial. SETTING AND PARTICIPANTS: The campaign was initiated in 4 communities in the greater Seattle, Wash, area between January 2000 and March 2001. Eight communities in Portland, Ore, and Spokane, Wash, served as control sites. We observed 3609 booster eligible children (those aged 4-8 years and weighing 18-36 kg [40-80 lb]). MAIN OUTCOME MEASURE: Observed booster seat use 15 months after the start of the campaign. RESULTS: Before the campaign began, 13.3% of eligible children in the intervention communities and 17.3% in the control communities were using booster seats, adjusting for child age, driver seat belt use, and sex of driver. Fifteen months after the start of the campaign, adjusted booster seat use had increased to 26.1% in the intervention communities and 20.2% in the control communities (P =.008 for the difference in time trends between intervention and control communities). CONCLUSION: These data suggest that a multifaceted community education campaign can significantly increase the use of child booster seats. PMID- 12588273 TI - Antibiotic resistance among gram-negative bacilli in US intensive care units: implications for fluoroquinolone use. AB - CONTEXT: Previous surveillance studies have documented increasing rates of antimicrobial resistance in US intensive care units (ICUs) in the early 1990s. OBJECTIVES: To assess national rates of antimicrobial resistance among gram negative aerobic isolates recovered from ICU patients and to compare these rates to antimicrobial use. DESIGN AND SETTING: Participating institutions, representing a total of 43 US states plus the District of Columbia, provided antibiotic susceptibility results for 35 790 nonduplicate gram-negative aerobic isolates recovered from ICU patients between 1994 and 2000. MAIN OUTCOME MEASURES: Each institution tested approximately 100 consecutive gram-negative aerobic isolates recovered from ICU patients. Organisms were identified to the species level. Susceptibility tests were performed, and national fluoroquinolone consumption data were obtained. RESULTS: The activity of most antimicrobial agents against gram-negative aerobic isolates showed an absolute decrease of 6% or less over the study period. The overall susceptibility to ciprofloxacin decreased steadily from 86% in 1994 to 76% in 2000 and was significantly associated with increased national use of fluoroquinolones. CONCLUSIONS: This study documents the increasing incidence of ciprofloxacin resistance among gram negative bacilli that has occurred coincident with increased use of fluoroquinolones. More judicious use of fluoroquinolones will be necessary to limit this downward trend. PMID- 12588274 TI - Medical monitoring for pharmaceutical injuries: tort law for the public's health? AB - A remarkable development in personal injury litigation in recent years involves attempts to expand legal claims beyond existing injuries to anticipated future harms. Attorneys have begun to sue on behalf of individuals exposed to defective pharmaceutical products who have no current injury, but who may be at risk for developing one after a latency period. This strategy seeks to make drug manufacturers pay for medical monitoring, a court-ordered program that provides diagnostic tests to exposed individuals to facilitate early detection of adverse health effects. Because medical monitoring does not depend on the existence of an actual injury and large populations may be exposed, some commentators have warned that it has the potential to spiral out of control. We examine medical monitoring in the context of 2 major cases involving diet drugs and an oral hypoglycemic drug. We conclude that this expansion of tort law should be applied sparingly, but that the performance of courts to date in these cases gives cause for optimism. Judges appear to be paying close attention to sophisticated epidemiological, clinical, and cost-effectiveness considerations. Medical monitoring arms the courts with a new mechanism for addressing harms proactively rather than reactively, which could yield new victories for public health. PMID- 12588275 TI - Is this woman perimenopausal? AB - CONTEXT: Perimenopause is a time of transition for women at midlife. Women want to know whether they are starting this change and physicians need to know the accuracy of a clinical examination in identifying perimenopausal women. These women should be counseled about alleviating climacteric symptoms, using contraception, and preventing diseases such as osteoporosis. OBJECTIVE: To systematically review the accuracy of self-assessment, symptoms, signs, and laboratory tests in diagnosing women in perimenopause. DATA SOURCES: English language articles that presented data relevant to diagnosis of perimenopause were identified in a MEDLINE search from 1966 to 2001. References of these articles and other publications also were reviewed. STUDY SELECTION: Cross-sectional or longitudinal studies of women aged 40 years or older that used the definition of perimenopause as 3 to 11 months of amenorrhea or irregular periods, included a premenopausal control group, and reported a clinical examination finding. Of 1246 articles identified, 16 studies were included in the analysis. DATA EXTRACTION: Two authors independently reviewed articles for quality (L.A.B. and C.M.S.). Discrepancies were resolved by a third author (K.N.). DATA SYNTHESIS: The prior probability of perimenopause is directly related to a woman's age. After considering age, the following yielded the greatest positive likelihood ratios (LRs+): self assessment of going through the transition (LR+ range, 1.53-2.13), symptoms of hot flashes (LR+ range, 2.15-4.06), night sweats (LR+ 1.90; 95% confidence interval [CI], 1.63-2.21), vaginal dryness (LR+ range, 1.48-3.79), high follicle-stimulating hormone levels (LR+ 3.06; 95% CI, 2.06-4.54), and low inhibin B levels (LR+ 2.05; 95% CI, 0.96-4.39). Self-assessment of perimenopausal status had the smallest negative LR (range, 0.18-0.36). CONCLUSIONS: No one symptom or test is accurate enough by itself to rule in or rule out perimenopause. Clinicians should diagnose perimenopause based on menstrual history and age without relying on laboratory test results. PMID- 12588276 TI - Should bivalirudin replace heparin during percutaneous coronary interventions? PMID- 12588277 TI - Managed care and patients' rights. PMID- 12588278 TI - The origins, history, and design of the resident match. AB - In the early 1900s, competition among hospitals for interns and among medical students for good internships led to increasingly early offers of internships to students. By the 1940s, appointments were often made as early as the beginning of the junior year of medical school. Hospitals thus had little information about students' performance, and students frequently had to make a final decision to accept or reject an offer without knowing which other offers might be forthcoming. From 1945 through 1951, efforts were made to enforce a uniform date for accepting offers. However, students were still faced with offers having very short deadlines, compelling them to accept or reject offers without knowing what other offers might be forthcoming. Hospitals often had to scramble for available students, since if an offer was rejected, it was often too late for them to reach their next preferred candidate. A centralized clearinghouse was thus developed as a way of alleviating this chaos and allowing a larger role to the preferences of both students and hospitals. This evolved into the current matching program, whose algorithm continues to be updated to take account of changing needs of applicants, such as growth in the number of couples who seek 2 positions in the same vicinity. PMID- 12588280 TI - JAMA patient page. Perimenopause: beginning of menopause. PMID- 12588279 TI - The National Resident Matching Program and antitrust law. AB - In May 2002, a group of physicians filed a class action lawsuit alleging that the National Resident Matching Program (NRMP) violates antitrust laws. The plaintiffs contend that NRMP practices have stabilized lower-than-competitive wages and imposed exhausting working conditions on residents. They also maintain that NRMP procedures virtually force applicants for house officer positions to forfeit their right to negotiate for better wages and conditions. The plaintiffs also allege that the defendants have collectively fostered anticompetitive accreditation standards through the Accreditation Council for Graduate Medical Education. Jung v Association of American Medical Colleges could present antitrust law with some difficult challenges. Although the matching program on its face appears to limit competition in a manner that previous cases have found illegal, it operates in the context of important professional activities (medical education and quality improvement) that may generate some judicial deference. At this early stage, no confident prediction can be made about the outcome of the case if it goes to trial; however, the plaintiffs appear to have a plausible case under existing antitrust doctrine, and lengthy litigation is possible. Given the important questions that the litigation will not address, such as the potential costs of a finding of illegality to the government and other payers, and the impact of such a finding on the health care system as a whole, a legislative solution seems highly desirable. PMID- 12588281 TI - Effects of adrenaline and potassium on QTc interval and QT dispersion in man. AB - BACKGROUND: Hypoglycaemia alters cardiac repolarization acutely, with increases in rate-corrected QT (QTc) interval and QT dispersion (QTd) on the electrocardiogram (ECG); such changes are related to the counterregulatory sympatho-adrenal response. Adrenaline produces both QTc lengthening and a fall in plasma potassium (K+) when infused into healthy volunteers. Hypokalaemia prolongs cardiac repolarization independently however, and therefore our aim was to determine whether adrenaline-induced repolarization changes are mediated directly or through lowered plasma K+. MATERIALS AND METHODS: Ten healthy males were studied on two occasions. At both visits they received similar l-adrenaline infusions but on one occasion potassium was also administered; infusion rates were adjusted to maintain circulating K+ at baseline. The QTc interval, QTd, peripheral physiological responses and plasma adrenaline and potassium concentrations were measured during both visits. RESULTS: The QTc interval and QTd increased both with and without potassium clamping. Without K+ replacement, mean (SE) QTc lengthened from 378 (5) ms to a final maximum value of 433 (10) ms, and QTd increased from 36 (5) ms to 69 (8) ms (both P < 0.001). During K+ replacement, QTc duration at baseline and study end was 385 (7) ms and 423 (11) ms, respectively (P < 0.001), and QTd 38 was (4) ms and 63 (5) ms (P = 0.001). CONCLUSIONS: These data suggest that disturbed cardiac repolarization as a result of increases in circulating adrenaline occurs independently of extracellular potassium. A direct effect of adrenaline upon the myocardium appears the most likely mechanism. PMID- 12588282 TI - Functional approach to investigate Lp(a) in ischaemic heart and cerebral diseases. AB - BACKGROUND: Lp(a), a major cardiovascular risk factor, contains a specific apolipoprotein, apo(a), which by virtue of structural homology with plasminogen inhibits the formation of plasmin, the fibrinolytic enzyme. A number of clinical reports support the role of Lp(a) as a cardiovascular or cerebral risk factor, and experimental data suggest that it may contribute to atherothrombosis by inhibiting fibrinolysis. DESIGN: A well-characterized model of a fibrin surface and an apo(a)-specific monoclonal antibody were used to develop a functional approach to detect pathogenic Lp(a). The assay is based on the competitive binding of Lp(a) and plasminogen for fibrin, and quantifies fibrin-bound Lp(a). High Lp(a) binding to fibrin is correlated with decreased plasmin formation. In a transversal case-control study we studied 248 individuals: 105 had a history of ischaemic cardiopathy (IC), 52 had cerebro-vascular disease (CVD) of thrombotic origin, and 91 were controls. RESULTS: The remarkably high apo(a) fibrin-binding in CVD (0.268 +/- 0.15 nmol L-1) compared with IC (0.155 +/- 0.12 nmol L-1) suggests the existence of peculiar and poorly understood differences in pro- or anti-thrombotic mechanisms in either cerebral and/or coronary arteries. CONCLUSIONS: Our results demonstrated that Lp(a) fibrin-binding and small Apo(a) isoforms are associated with athero-thrombotic disease. PMID- 12588283 TI - Vitamin D receptor gene polymorphism BsmI is not associated with the prevalence and severity of CAD in a large-scale angiographic cohort of 3441 patients. AB - BACKGROUND: Recent studies found a relationship between Vitamin D and atherosclerosis. A common genetic polymorphism of the Vitamin D receptor (VDR) has been associated with coronary artery disease (CAD) in small study populations. To assess its influence on the prevalence and severity of CAD we studied a large-scale population. METHODS: A total of 3441 consecutive patients were referred for diagnostic coronary angiography. The BsmI Vitamin D receptor polymorphism was analyzed by polymerase chain reaction. Angiography was used to define phenotypes with clear coronary arteries (n = 775), coronary sclerosis (diameter stenosis < 50%; n = 579), CAD (diameter stenosis > 50% in at least one vessel; n = 1524). Patients with CAD at a young age (females aged less than 65 years, males aged less than 55 years; n = 563) were specially defined as premature CAD. The risk profile of traditional cardiovascular risk factors was obtained for every patient. RESULTS: The genotype frequencies of the VDR BsmI polymorphism did not differ between all four phenotypes (P = 0.756). The allele frequencies for the B allele were 0.43 vs. 0.44 vs. 0.42 vs. 0.45 in the four phenotypic groups (P = 0.827). All traditional cardiovascular risk factors (hypercholesterolaemia, smoking, hypertension, diabetes mellitus, severe obesity, male gender) were significantly (P < 0.001) associated with the angiographic phenotype. CONCLUSIONS: The VDR gene variant BsmI was not associated with prevalence and severity of CAD in a large-scale cohort phenotyped by angiography. PMID- 12588284 TI - Intima media thickness of carotid arteries is reduced in heterozygous carriers of the Gly972Arg variant in the insulin receptor substrate-1 gene. AB - BACKGROUND: The Gly972Arg mutation in the IRS-1 gene has been found to be associated with insulin resistance and type II diabetes. A recently published study described an association between the Arg allele and an increased risk for coronary artery disease. In the present study we asked whether the presence of the codon 972 mutation in the IRS-1 gene is associated with higher IMT values of the carotid arteries. MATERIALS AND METHODS: To address this question, genotypes of the codon 972 polymorphism were determined in 1018 healthy unrelated individuals aged 40-65 years. Three homozygous carriers of the mutation were excluded for statistical analysis. In all subjects, intima media thickness (IMT) and B-scores of carotid arteries as well as a large number of metabolic parameters were determined. RESULTS: Heterozygous carriers of the Arg972 allele exhibited significantly lower IMT and B-score values than noncarriers. Total cholesterol, LDL-cholesterol and serum levels of apolipoprotein B were significantly lower in the carriers. Furthermore, a significant interaction between Gly972Arg-carrier status and mean daytime 24-h systolic blood pressure with regard to IMT could be observed; carriers with a systolic blood pressure above the median had lower IMT values than carriers with a systolic blood pressure equal or below the median. All these effects were more pronounced in females and remained significant after adjustment for sex, age, BMI, systolic blood pressure and serum apolipoprotein B levels. No significant differences between the carriers and the noncarriers could be found for BMI, insulin sensitivity or frequency of type II diabetes. CONCLUSIONS: The results of our study demonstrate that the presence of the Arg972 allele is associated with lower IMT values of the carotid arteries. This finding is partly explained by lower serum levels of apolipoprotein B in carriers. The protective effect of the Gly972 Arg mutation seems to be stronger in the presence of a higher systolic blood pressure. Our data contradict previous findings suggesting an increased risk for insulin resistance, type II diabetes and atherosclerotic vascular disease in carriers of the mutation. PMID- 12588285 TI - The effect of lipoproteins on endothelial nitric oxide synthase is modulated by lipoperoxides. AB - BACKGROUND AND AIM: The effect of low-density lipoproteins (LDLs) on endothelial nitric oxide synthase (eNOS) is debated. By coupling in vivo and in vitro experiments we evaluated the role of oxidized lipid substrates in the modulation of eNOS activity by LDLs. MATERIALS AND METHODS: Plasma lipids, nitrite/nitrates (NO2/NO3), and malondialdehyde (MDA) were measured in 14 controls, and in 13 patients with familial hypercholesterolemia (FH) before and after 12 weeks of treatment with atorvastatin (20 mg u.i.d.). Nitric oxide synthase in cell lysate and NO2/NO3 into the medium were measured in human microvascular (HMEC-1) and umbilical vein (HUVEC) endothelial cells after 24 h of incubation with increasing concentrations of mildly oxidized LDLs with and without atorvastatin and in HMEC 1 with and without vitamin C. In HMEC-1, NO2/NO3 was also determined after exposure to more intensively oxidized LDLs. RESULTS: At baseline, plasma NO2/NO3 (56 +/- 7 vs. 35 +/- 3 micro M) and MDA (5.6 +/- 0.7 vs. 2.9 +/- 0.3 micro M), were significantly (P < 0.02 for both) higher in the FH patients. In the whole study group, NO2/NO3 was more strongly correlated with plasma MDA (Rho = 0.70) than LDL-cholesterol (Rho = 0.57). In the FH patients, atorvastatin induced a significant decline in plasma total and LDL-cholesterol (-3.1 +/- 0.5 and -2.9 +/ 0.5 mM, respectively), NO2/NO3 (-35 +/- 8 microM) and MDA (-3.4 +/- 0.7 microM) (P < 0.001 for all). Changes in plasma NO2/NO3 were related to the concomitant changes in plasma MDA (Rho = 0.79, P < 0.006) and not to changes in LDL cholesterol. In HMEC-1 and in HUVEC, mildly oxidized LDLs stimulated both e-NOS and NO2/NO3 accumulation; the effect on e-NOS was potentiated by vitamin C in HMEC-1. Atorvastatin had no effect in HMEC-1 while it stimulated eNOS but not NO2/NO3 in HUVEC. The accumulation of NO2/NO3 in HMEC exposed to increasing concentrations of more intensively oxidized-LDLs showed a nonlinear dose-response curve. CONCLUSIONS: In uncomplicated patients with FH, plasma NO2/NO3 concentrations are elevated; the cross-sectional data, intervention study and in vitro experiments indicate that oxidized lipids exert a tonic stimulatory action on e-NOS and NO2/NO3 generation not mediated through superoxide anion formation. Atorvastatin amplify this eNOS response in HUVEC, but not in HMEC, and this effect is not associated with a parallel increased NO2/NO3 generation. PMID- 12588286 TI - Low-density-lipoprotein peak particle size in a Mediterranean population. AB - BACKGROUND: The predominance of small, dense low-density lipoprotein (LDL) particles ('LDL phenotype B') has been associated with a three-fold increased risk of myocardial infarction, but the feasibility of the identification of small, dense LDL as independent predictors of coronary artery disease risk in population studies remains questioned. Design We evaluated the LDL peak particle size and its relation with other established risk factors for coronary heart disease in a group of 156 randomized subjects living on the Mediterranean island of Ustica (71 males and 85 women, range of age 20-69 years), representing approximately 30% of the total population. RESULTS: The prevalence of LDL phenotype B subjects was low (approximately 15% in both men and women) and there was a clear trend for both genders in reducing the LDL peak particle size with age. Moreover, LDL phenotype B subjects had higher BMI values, prevalence of diabetes and plasma triglyceride (TG) levels and lower plasma HDL-C concentrations in comparison with LDL phenotype A individuals; in a multivariate analysis, plasma TG levels were the only variable independently associated with LDL peak particle size. CONCLUSIONS: In this population, which appears to be somewhat protected by premature coronary artery disease, a low prevalence of the LDL pattern B was found in both men and women, and plasma TG could have a key role in regulating the LDL peak particle size. The follow up, still ongoing, will provide useful information on the predictive role of LDL peak particle size on cardiovascular risk, at least in a low-risk population. PMID- 12588287 TI - No acute impact of lipid apheresis treatment on free radical scavenging enzyme gene expression in white blood cells. AB - BACKGROUND: Lipid apheresis (LA) treatment has been suggested to cause oxidative stress. Defense against oxygen-radical-mediated damage is provided by nonenzymatic and enzymatic antioxidants. In the present investigation we have investigated whether gene expression of free radical scavenging enzymes (FRSE) is affected in leukocytes of patients undergoing LDL-apheresis. MATERIALS AND METHODS: For this purpose cellular glutathione peroxidase (GPx-1), phospholipid glutathione peroxidase (GPx-4), glutathione reductase (GSSG-R), glutathione synthetase (GSH-S), Cu/Zn-superoxide dismutase (SOD-1) and catalase (CAT) mRNA expression were followed at the start (SA) and immediately after (EA) LA treatment (n = 25). Gene expression was determined by quantitative RT-PCR with the LightCycler(R) instrument (Roche Diagnostics, Mannheim, Germany) and transcription elongation factor-2 as reference gene. RESULTS: The expression of GPx-1, GPx-4, GSSG-R, GSH-S, SOD-1, CAT mRNA was not affected by a single LA treatment. Free radical scavenging enzymes mRNAs were significantly (P < 0.05) increased in the LA patients (GPx-1: 2.00 +/- 1.37; GPx-4: 0.52 +/- 0.46; GSSG-R: 0.07 +/- 0.03; GSH-S: 0.04 +/- 0.03; SOD-1: 1.12 +/- 0.74; CAT: 0.15 +/- 0.07) when compared with 26 healthy blood donors (GPx-1: 1.1 +/- 0.6; GPx-4: 0.35 +/- 0.19; GSSG-R: 0.02 +/- 0.01; GSH-S: 0.03 +/- 0.01; SOD-1: 0.16 +/- 0.08; CAT: 0.09 +/- 0.05; mean +/- SD). CONCLUSIONS: These results show that the LA procedure does not acutely affect the antioxidant defense system on the gene level but suggests that the chronic stress resulting from hyperlipidaemia and/or LA may cause FRSE gene induction. PMID- 12588288 TI - Transcapillary insulin transfer in human skeletal muscle. AB - BACKGROUND: Transcapillary insulin transfer is considered a rate-limiting step in insulin action at supraphysiological insulin concentrations. However, it remains unclear whether this concept also applies for physiological conditions. MATERIALS AND METHODS: In the present study we set out to characterize transcapillary insulin transfer by measuring insulin concentrations in plasma and interstitial space fluid of skeletal muscle during an oral glucose tolerance test and euglycaemic hyperinsulinaemic clamp conditions, respectively. For this purpose we employed in vivo microdialysis of skeletal muscle in conjunction with an ultrasensitive insulin assay in eight healthy lean male volunteers (aged 25 +/- 1 years). RESULTS: Insulin concentrations at baseline were 48 +/- 8 pmol x L(-1) in plasma and 19 +/- 4 pmol x L(-1) in the interstitium (P = 0.002). The mean interstitium to plasma ratio at baseline was 0.48 +/- 0.09 pmol x L(-1). During the oral glucose tolerance test the interstitium to plasma ratio remained unchanged (0.43 +/- 0.12, P = NS vs. baseline), but was significantly reduced during euglycaemic hyperinsulinaemic clamp conditions at steady-state hyperinsulinaemia (0.12 +/- 0.01, P = 0.01 vs. baseline). CONCLUSION: In summary there is a substantial transcapillary insulin gradient in healthy human skeletal muscle under baseline and glucose-stimulated conditions. Our findings support the hypothesis of a saturable transcapillary insulin transport representing a partly rate-limiting step for insulin action. PMID- 12588290 TI - Mouse models for genetic dissection of polygenic gastrointestinal diseases. AB - Many diseases with a major public health impact are the result of complex interactions between environmental factors and multiple genes. In the past decade, methods for genome analysis, in particular quantitative trait locus (QTL) analysis in animal models, were developed to identify and localize the genes responsible for multifactorial (polygenic) diseases; QTL analysis is based on experimental crosses between inbred strains with high and low genetic susceptibility. Recently the genes underlying several QTLs could be cloned successfully. Here we describe the impact of these genomic approaches in mice on our understanding of the multifactorial genetics of three gastrointestinal diseases related to metabolism (cholesterol cholelithiasis), development (gastroschisis), and colorectal cancer. The examples demonstrate how mouse models continue to be an invaluable tool in unravelling complex pathomechanisms and unlocking our understanding of human diseases. PMID- 12588289 TI - Ratio between serum IL-8 and pepsinogen A/C: a marker for atrophic body gastritis. AB - BACKGROUND AND AIMS: Elevated serum gastrin and a low pepsinogen A/C ratio are well-recognized markers for atrophic body gastritis (ABG). We have shown that the presence of body atrophy is also associated with elevated serum pro-inflammatory cytokines. This study tested the hypothesis that serum cytokines provide additional information to gastrin and pepsinogens in screening for ABG. METHODS: Two hundred and twenty-six consecutive patients were investigated on referral for upper gastrointestinal endoscopy: 150 were patients with gastro-oesophageal reflux disease, receiving acid inhibitory medication either with proton pump inhibitors (n = 113) or with histamine2-receptor antagonists (n = 37), and 76 were nontreated controls, who had normal endoscopic findings. Gastric mucosal biopsies were sampled for histological examination (Sydney classification). Serum samples were analyzed for gastrin, chromogranin A (CgA), and pepsinogens A and C by RIA, and for the interleukins (IL)-1beta, IL-6, and IL-8 by ELISA. RESULTS: Subjects with ABG had significantly higher serum gastrin (P < 0.01) and serum CgA (P < 0.01) levels and significantly lower pepsinogen A/C ratios (P < 0.001) than those without ABG. Additionally, serum IL-1beta, IL-6 and, especially, IL-8 levels were significantly higher in the subjects with than in those without ABG (P < 0.0001, for all cytokines). To optimize the detection of body atrophy we defined the ABG index: the ratio between the simultaneously measured IL-8 and pepsinogen A/C. The area under the ROC curve for the ABG index was significantly greater than that for serum gastrin and for serum pepsinogen A/C alone (0.91 +/- 0.029 vs. 0.72 +/- 0.042, and vs. 0.83 +/- 0.031, P = 0.018 and P = 0.049). Using the ABG index at a cut-off value of 1.8 pg mL-1, 91% of the cases were classified correctly. CONCLUSIONS: The ratio between serum IL-8 and pepsinogen A/C accurately predicts the presence of ABG. We therefore propose the ABG index as a noninvasive screening test for ABG in population-based studies. PMID- 12588291 TI - Anti-cardiolipin antibodies in patients with chronic viral hepatitis are independent of beta2-glycoprotein I cofactor or features of antiphospholipid syndrome. AB - BACKGROUND: Although controversial, some authorities have implicated hepatitis C virus (HCV) as a cause of anti-phospholipid syndrome (APLS). Anti-cardiolipin antibodies (anti-CLAbs) in APLS are cofactor-dependent ('pathogenic' antibodies). We conducted a study in order to determine the prevalence of anti-CLAbs in HCV patients, and furthermore to address whether these autoantibodies are cofactor dependent or not and whether they are associated with features of APLS. Patients with hepatitis B virus (HBV) were also evaluated in order to assess whether there are differences in the prevalence and the clinical significance of anti-CLAbs between these two major types of chronic viral hepatitis. MATERIALS AND METHODS: One hundred and seventy-four consecutive HCV patients, 50 HBV patients and 267 healthy were investigated for the presence of anti-CLAbs and antibodies against beta2-glycoprotein I (beta2-GPI), which is the most important cofactor of the 'pathogenic' anti-CLAbs in APLS. IgG anti-CLAbs were determined by an in-house quantitative ELISA and anti-beta2-GPIAbs using a commercial ELISA kit. RESULTS: 21.3% of the HCV and 14% of the HBV patients tested positive for IgG anti-CLAbs (P < 0.0001 compared with healthy controls). Neither age, sex, certain epidemiologic and laboratory parameters nor the clinical status and the histologic findings were associated with anti-CLAbs detection in both diseases. 2.3% of the HCV (P < 0.05 compared with healthy controls) and 2% of the HBV patients tested positive for anti-beta2-GPIAbs. Presence of anti-CLAbs was not associated with features of APLS. CONCLUSIONS: A significant proportion of the HCV and HBV patients had detectable IgG anti-CLAbs. However, the anti-CLAbs titres were relatively low, and in most cases seem to be cofactor-independent ('nonpathogenic'). The latter is further supported by the lack of their association with clinical features of APLS. Furthermore, anti-CLAbs appear to be detected irrespective of the demographic, laboratory, clinical and histologic status in both HCV and HBV. However, prospective studies of longer duration may be required in order to address whether anti-CLAbs in patients with chronic viral hepatitis are or are not of clinical importance. PMID- 12588292 TI - Pneumococcal immune adherence to human erythrocytes. AB - BACKGROUND: Human red blood cells bind various C3b-coated microorganisms via their C3b/CR1 receptor, a phenomenon referred to as immune adherence. The aim of the present study was to measure pneumococcal adherence to human red blood cells by flow cytometry and to study kinetic aspects of this binding. MATERIAL AND METHODS: We quantified pneumococcal adherence to human erythrocytes by FACS analysis and tested the involvement of antibodies and complement activation in this process. RESULTS: Pneumococci are able to bind to human red blood cells in the presence of human serum. Coating with C3b/C4b appeared obligatory for pneumococcal adherence to red blood cells. The ligand on erythrocytes was confirmed to be complement receptor 1. Kinetic studies showed that innate (mannose-binding lectin) and specific immune factors (IgG antibodies) contributed to the binding of C3b-coated pneumococci to human erythrocytes. After initial binding, serum-derived factor I was found to induce bacterial detachment from the erythrocyte. CONCLUSIONS: Pneumococci are able to adhere to red blood cells. Both the classical and lectin complement pathways are important for optimal C3b coating of pneumococci for immune adherence. Bound pneumococci are detached from red blood cells by factor I. These findings are in line with the hypothesis of immune adherence in which human erythrocytes are able to bind pneumococci and target the bacteria to the reticulo-endothelial system in the spleen. PMID- 12588293 TI - Intracellular glutathione and lipid peroxide availability and the secretion of vasoactive substances by human umbilical vein endothelial cells after incubation with TNF-alpha. AB - BACKGROUND: The major pathophysiologic changes observed in preeclampsia suggest that endothelial cell dysfunction plays an important role in this disorder. The pathway mediating to endothelial cell dysfunction is unknown, however, the pathogenesis of preeclampsia is thought to be related to increased oxidative stress and increased vasoconstriction. The concentration of tumor necrosis factor alpha (TNF-alpha), a cytokine produced by macrophages and many other cell types, has been observed to be significantly increased in preeclampsia. It has been hypothesized that TNF-alpha overproduction by the placenta may then may produce an increase in plasma levels and subsequent endothelial dysfunction in preeclampsia. This study investigated the effect of TNF-alpha on glutathione and lipid peroxide levels and on the secretion of vasoactive substances by human umbilical vein endothelial cells (HUVECs). METHODS: Human umbilical vein endothelial cells were incubated for 24 h in the presence of different concentrations of TNF-alpha (0-1000 pg mL-1) that were shown in an earlier experiment to have no effects on the vitality and proliferation rate of HUVECs. The levels of reduced glutathione (GSH) and lipid peroxides (LPOs), assessed by malondialdehyde and 4-hydroxyalkenal, were measured in endothelial cell lysates. For the measurement of vasoactive substances, levels of prostacyclin (PGI2), determined by 6-keto-prostaglandin F1a, thromboxane A2 (TXA2), measured by thromboxane B2, endothelin-1 (ET-1), and nitric oxide (NO), measured by total nitrite, were assessed in endothelial cell supernatants. RESULTS: At lower concentrations (10-100 pg mL-1), TNF-alpha increases the intracellular content of LPO and GSH, stimulates the secretion of ET-1 and TXA2, but inhibits the secretion of PGI2 in endothelial cells compared with control cells. At concentration of 1000 pg mL-1, TNF-alpha increases the secretion of PGI2 and TXA2, but it decreases the ET-1 concentration. TNF-alpha has no effect on NO secretion. CONCLUSION: These findings demonstrate that at concentrations corresponding to values in plasma from preeclamptic women, TNF-alpha induces oxidative stress and results in altered secretion of vasoactive substances in favour of vasoconstrictors in human endothelial cells. We conclude that TNF-alpha may participate in the pathway leading to endothelial cell dysfunction seen in preeclampsia. PMID- 12588294 TI - Bacteria and Archaea Group II introns: additional mobile genetic elements in the environment. AB - Self-splicing group II introns are present in the organelles of lower eukaryotes, plants and Bacteria and have been found recently in Archaea. It is generally accepted that group II introns originated in bacteria before spreading to mitochondria and chloroplasts. These introns are thought to be related to the progenitors of spliceosomal introns. Group II introns are also mobile genetic elements. In bacteria, they appear to spread using either other mobile genetic elements or low-expression regions as target sites. Bacteria and Archaea genome sequence annotations have revealed the diversity of group II intron classes and that they are involved in vertical and horizontal inheritance. PMID- 12588295 TI - The impact of grassland management on archaeal community structure in upland pasture rhizosphere soil. AB - The community structure of rhizosphere soil Archaea from three grassland types, associated with different management practices, was examined at a site in the Borders region of Scotland, by analysis of 16S rRNA gene fragments amplified from 16S rDNA and from rRNA. Denaturing gradient gel electrophoresis (DGGE) and sequence analysis of amplified products indicated high relative abundance within the archaeal community of two distinct lineages of non-thermophilic (group 1) Crenarchaeota. Grassland management practices influenced archaeal community structure, as characterized by both 16S rRNA- and 16S rDNA-derived DGGE profiles. One band dominated DGGE profiles in all three grassland types examined, and reproducible differences in the presence and intensity of bands were observed between profiles from managed and natural grassland sites. Analysis of 16S rRNA derived amplicons from managed and natural grasslands at sites in the north of England and the north of Wales also indicated high relative abundance of non thermophilic crenarchaeotes within the archaeal community. The band dominating the Scottish grassland site also dominated DGGE profiles from the English and Welsh sites, and similar differences were seen between profiles derived from soils subjected to different management regimes. The study indicates that grassland archaeal communities are dominated by Crenarchaeota, with closely related members of this lineage ubiquitous in distribution in UK upland pasture, and indicate that management practices influence the nature of the crenarchaeotal community. PMID- 12588296 TI - Evolution of a chlorobenzene degradative pathway among bacteria in a contaminated groundwater mediated by a genomic island in Ralstonia. AB - The genetic structure of two Ralstonia spp., strain JS705 and strain JS745, isolated from the same groundwater aquifer, was characterized with respect to the degradation capacities for toluene and chlorobenzene degradation. Cosmid library construction, cloning, DNA sequencing and mating experiments indicated that the genes for chlorobenzene degradation in strain JS705 were a mosaic of the clc genes, previously described for Pseudomonas sp. strain B13, and a 5 kb fragment identical to strain JS745. The 5 kb fragment identical to both JS705 and JS745 was flanked in JS705 by one complete and one incomplete insertion (IS) element. This suggested involvement of the IS element in mobilizing the genes from JS745 to JS705, although insertional activity of the IS element in its present configuration could not be demonstrated. The complete genetic structure for chlorobenzene degradation in strain JS705 resided on a genomic island very similar to the clc element (Ravatn, R., Studer, S., Springael, D., Zehnder, A.J., van der Meer, J.R. 1998. Chromosomal integration, tandem amplification, and deamplification in Pseudomonas putida F1 of a 105-kilobase genetic element containing the chlorocatechol degradative genes from Pseudomonas sp. strain B13. J Bacteriol 180: 4360-4369). The unique reconstruction of formation of a metabolic pathway through the activity of IS elements and a genomic island in the chlorobenzene-degrading strain JS705 demonstrated how pathway evolution can occur under natural conditions in a few 'steps'. PMID- 12588297 TI - Cloning of Baeyer-Villiger monooxygenases from Comamonas, Xanthobacter and Rhodococcus using polymerase chain reaction with highly degenerate primers. AB - To clone novel type 1 Baeyer-Villiger monooxygenase (BVMO) genes, we isolated or collected 25 bacterial strains able to grow on alicyclic compounds. Twelve of the bacterial strains yielded polymerase chain reaction (PCR) fragments with highly degenerate primers based on the sequences of known and putative BVMOs. All these fragments were found to encode peptides homologous to published BVMO sequences. The complete BVMO genes and flanking DNA were cloned from a Comamonas, a Xanthobacter and a Rhodococcus strain using the PCR fragments as probes. BVMO genes cloned from the first two strains could be expressed to high levels in Escherichia coli using standard expression vectors, and the recombinants converted cyclopentanone and cyclohexanone to the corresponding lactones. The Rhodococcus BVMO, a putative steroid monooxygenase, could be expressed after modification of the N-terminal sequence. However, recombinants expressing this protein did not show activity towards progesterone. An esterase homologue located directly upstream of the Xanthobacter BVMO gene and a dehydrogenase homologue encoded directly downstream of the Comamonas sp. NCIMB 9872 BVMO gene were also expressed in E. coli and shown to specify lactone hydrolase and cyclohexanol dehydrogenase activity respectively. PMID- 12588298 TI - Microbial community and physicochemical analysis of an industrial waste gas biofilter and design of 16S rRNA-targeting oligonucleotide probes. AB - A study was conducted to investigate the microbial community structure, the physicochemical properties, and the relationships between these parameters of a full-scale industrial biofilter used for waste gas abatement in an animal rendering plant. Fluorescence in situ hybridization (FISH) was successfully combined with digital image analysis to study the composition of the microbial community. Several new nucleic acid probes were designed and established based on published 16S rDNA sequences and on ones retrieved from the biomass of the biofilter under investigation. Bacterial detection rates varied greatly over time and filterbed depth between 27.2% and 88.1% relative to DAPI counts. Overall, members of the Betaproteobacteria followed by Actinobacteria, Alphaproteobacteria, Cytophaga-Flavobacteria, Firmicutes and Gammaproteobacteria were the most abundant groups. Among the groups below phylum level, members of the Alcaligenes/Bordetella lineage were on average the most abundant group accounting for up to 8.5% of DAPI-stained cells. Whereas the community composition generally showed no vertical gradient, the lower 50 cm of the biofilter proved to be the most active part for the degradation of aldehydes such as 2- and 3-methylbutanal, 2-methylpropanal, and hexanal. This zone of the filterbed being operated in up-flow direction degraded about 80% of these compounds. Dimethyldisulphide was the most common reduced sulphur compound. Statistical analysis of microbial versus waste gas parameters generally revealed only weak or non-significant correlations between the two. Possible explanations for this finding are discussed. PMID- 12588299 TI - Quantification of cell-specific substrate uptake by probe-defined bacteria under in situ conditions by microautoradiography and fluorescence in situ hybridization. AB - A technique based on quantitative microautoradiography (QMAR) and fluorescence in situ hybridization (FISH) was developed and evaluated in order to determine the quantitative uptake of specific substrates in probe-defined filamentous bacteria directly in a complex system. The technique, QMAR-FISH, has a resolution of a single cell and is based on an improved fixation protocol and the use of an internal standard of bacteria with known specific radioactivity. The method was used to study the in situ ecophysiology of the filamentous bacteria 'Candidatus Meganema perideroedes' and Thiothrix sp. directly in an activated sludge system. The cellular uptake rate of tritium-labelled substrates revealed an average cell specific uptake rate of 4.1 yen 10-15 mol of acetate cell-1 h-1 and 3.1 yen 10-15 mol of acetate cell-1 h-1 for the two filamentous species respectively. The two filamentous species had very similar activity in all cells along each filament. Surprisingly, the filaments within both probe-defined populations had threefold variation in activity between the different filaments, demonstrating a large variation in activity level within a single population in a complex system. The substrate affinity (Ks) for uptake of acetate of the cells within the two filamentous bacteria was determined by incubation with variable concentrations of labelled acetate. The Ks values of the 'Candidatus Meganema perideroedes' and the Thiothrix filamentous bacteria were determined to be 1.8 micro M and 2.4 micro M acetate respectively. PMID- 12588300 TI - Molecular diversity among marine picophytoplankton as revealed by psbA analyses. AB - Photosynthetic microorganisms play a crucial role in the marine environment. In vast areas of the oceans, marine primary productivity is performed by cells smaller than 2-3 micro m (picoplankton). Here, we report on molecular analyses of the conserved photosynthetic psbA gene (coding for protein D1 of photosystem II reaction centre) as a diversity indicator of naturally occurring marine oxygenic picophytoplankton. The psbA genes proved to be good indicators of the presence of a wide variety of photosynthetic marine microbial groups, including new cyanobacterial groups and eukaryotic algae (prasinophytes). Furthermore, using environmental bacterial artificial chromosome (BAC) libraries, we were able to correlate psbA genes with small subunit rRNAs and, therefore, to confirm their phylogenetic affiliation. PMID- 12588303 TI - Informed decision making for cancer screening--not all of the ethical issues have been considered. PMID- 12588304 TI - Abnormal chromosome 8 copy number in cytological smears from breast carcinomas detected by means of fluorescence in situ hybridization (FISH). AB - Numerical change in chromosome 8 is an acquired abnormality associated with high clinical stage and may be involved in the conversion of carcinoma in situ in the breast to invasive carcinoma. Fine needle aspiration smears from 53 cases of breast carcinoma were hybridized with centromeric probes for chromosome 8 and the X chromosome. Thirty-eight cases revealed chromosome 8 copy gain. Of the 45 grade II and III tumours, 28 showed polysomy (>3 signals) and six showed trisomy. Of the eight grade I tumours, four were trisomic, none were polysomic. There were only two cases of chromosome 8 copy loss (one each of grade I and III). X chromosome polysomy was also a frequent finding although the signal counts were similar to those for chromosome 8 in only a few cases. Chromosome 8 polysomy occurs frequently in breast carcinoma and high copy number (>3) is associated with high malignancy grade. PMID- 12588305 TI - Cytology suggestive of glandular neoplasia: outcomes and suggested management. AB - Eighty-three cases having a cervical smear result showing abnormal glandular cells were identified and matched up with the diagnostic histology result. Thirty four (41.0%) were associated with malignancy and 26 (31.3%) with a cervical intraepithelial lesion without invasion. Thirty-eight (45.8%) had conditions of the cervix as follows: 12 cases had invasive disease of the cervix; nine (10.8%) adenocarcinoma of cervix and three (3.6%) squamous carcinoma of cervix. Nineteen (22.9%) had cervical intraepithelial neoplasia (CIN/SIL) alone and seven (8.4%) had cervical glandular intraepithelial neoplasia (CGIN) +/- CIN. There were 16 (19.3%) cases with malignancies of the uterine corpus and six (7.2%) had a malignancy arising from another primary site. Twenty-three (27.7%) had no malignant or pre-malignant condition. The risk of malignancy was related to age and ranged from 18.2% in those under 35 years to 67.9% in those 55 years and over. A protocol for the management of these cases is described. PMID- 12588306 TI - Cytology of abnormal nipple discharge: a cyto-histological correlation. AB - Spontaneous or expressible nipple discharge may occur in palpable and nonpalpable breast lesions. The aim of the study was to evaluate the sensitivity and specificity of nipple discharge cytology in palpable and nonpalpable breast lesions. One hundred and seventy-four nipple discharge specimens were reviewed, of which 82 had corresponding surgical pathology, including 34 palpable breast lesions and 48 nonpalpable breast lesions. There was good correlation between nipple discharge cytology and concomitant fine needle aspiration (FNA) cytology. Nipple discharge cytology is as specific as concomitant FNA cytology but is slightly less sensitive in detecting papillomas or malignant lesions. The sensitivity and specificity of the nonpalpable and palpable breast lesions were similar. Nipple discharge cytology is very helpful in detecting an underlying breast lesion even if the case has no palpable mass in the breast. PMID- 12588307 TI - Dipeptidyl peptidase IV expression in thyroid cytology: retrospective histologically confirmed study. AB - Fine needle aspiration cytology (FNAC) of the thyroid gland is a well-established method. However, it has inherent limitations, especially in the diagnosis of follicular and oncocytic tumours and in distinguishing between nuclear atypia in colloid goitre with regressive changes and cystic papillary carcinoma. The aim of our study was to evaluate dipeptidyl peptidase IV (DPP IV) as a marker of malignancy in FNAC. We tested 254 thyroid specimens (intraoperative imprint smears) for DPP IV. The sensitivity was 71%, the specificity was 96%, and the diagnostic accuracy was 93%, respectively, with a threshold of 50% of positive cells. To the best of our knowledge it is the largest histologically confirmed study reported in the literature. We suggest the assessment of DPP IV as an adjunct diagnostic marker of malignancy in thyroid specimens suspicious of papillary carcinoma. However, the value of the marker in follicular lesions is very limited. PMID- 12588308 TI - SM047 immunoreactivity in peritoneal fluids. AB - SM047 is a recently developed monoclonal antibody generated against an ovarian adenocarcinoma cell line. A recent immunohistochemical study has shown that SM047 is strongly expressed in tissue sections of most ovarian serous adenocarcinomas. This study aimed to ascertain whether SM047 staining is of value in cytological preparations of peritoneal fluid. A total of 206 consecutive peritoneal fluids were stained immunocytochemically with SM047, CA125, monoclonal carcinoembryonic antigen (mCEA), Ber-EP4 and cytokeratins (CK7 and 20). SM047 positivity was present in reactive mesothelial cells in 117 of 141 (83%) benign cases in which these were present. SM047 positive tumour cells were present in 22 of 23 (96%) ovarian serous adenocarcinomas and in small numbers of gastric adenocarcinomas (two of three), mesotheliomas (one of two) and pancreatic adenocarcinomas (one of one). All six colorectal and two breast adenocarcinomas were negative with SM047. Reactive mesothelial cells in all cases were positive with CK7 and in most cases with CA125. They were negative with CEA, Ber-EP4 and CK20. All adenocarcinomas were positive with Ber-EP4 and mesothelial cells were always negative. All colorectal adenocarcinomas were positive with CK20. This study shows that SM047 staining may be of value in the diagnosis of an ovarian serous adenocarcinoma in peritoneal fluids. Negative staining helps to exclude a primary ovarian serous adenocarcinoma and is characteristic of colorectal adenocarcinoma. The small numbers of other malignancies in the study precludes a judgement of the value of SM047 staining in these neoplasms. SM047 staining may be useful, as part of a larger panel, in the work up of patients with peritoneal effusions. PMID- 12588310 TI - Fine needle aspiration from the upper arm in a postmastectomy patient. PMID- 12588311 TI - Alveolar soft part sarcoma: a rare entity with unique cytomorphological features. PMID- 12588312 TI - Evaluation of liquid-based cytology in cervical screening of high-risk populations. PMID- 12588314 TI - Stroke in renal transplant recipients: epidemiology, predictive risk factors and outcome. AB - BACKGROUND: Cerebrovascular and cardiovascular diseases are the most important causes of increased morbidity and mortality in patients with end-stage renal disease. Stroke has been widely reported in chronic dialysis patients, but there is scarce information about stroke in renal transplant recipients (RTR), although cerebrovascular events are the most common and potentially life-threatening neurological complications in them. Our aim is to analyze the prevalence, risk factors, etiopathogenia, clinical aspects and outcome of stroke in RTR. METHODS: We analyzed 403 patients who received one or more renal grafts between 1979 and 2000: group A = patients who had stroke (n = 19); group B = those who did not (n = 384). Medical records and pertinent data were compiled. The risk of stroke was studied using univariate and multivariate Cox regression models. RESULTS: prevalence of stroke in RTR was 7.97% at 10 yr. Time elapsed between renal transplantation (RT) and stroke: 49.3 months. Possible risk factors based on the univariate analyses were: diabetic nephropathy (DN) (p < 0.001) and autosomal dominant-polycystic-kidney-disease (p = 0.049) as original nephropathies, peripheral vascular disease (PVD) (p < 0.001), diabetes mellitus prior to RT (p = 0.005), age older than 40 yr (p = 0.037) and hypertension (p = 0.049). Other analysed risk factors such as gender, renal function, cytomegalovirus infection, hyperlipidemia, hyperuricemia, erythrocytosis or hypertensive donor failed to show any significant predictive value for stroke in these patients. When multivariate analyses were carried out, we found that DN (OR = 4.8; p = 0.010), PVD (OR = 8.2; p < 0.001) and age > 40 yr (OR = 3.3; p = 0.019) were predictive risk factors for stroke. For group A, hypertension was present in all patients, 68.4% had hyperlipidemia and 42.1% reported previous stroke. Cerebral hemorrhage occurred in seven of 19 (36.84%) of the stroke patients, but no subarachnoid hemorrhage occurred in them. Seven of 12 ischemic strokes were atherotrombotic. Considering all strokes, basal ganglia was the predominant localization. The outcome was poor, as nearly half of the patients died in the 3 months following stroke. CONCLUSIONS: Prevalence of stroke in our RTR population was 7.97%. Cerebral hemorrhage appears to be more prevalent in RTR than in general population. More than that, the cerebral hemorrhage rate we found is higher than that reported elsewhere in RTR. The main predictors of stroke were DN, PVD and age. No patient with interstitial nephropathy suffered stroke. Mortality is high in RTR with stroke. PMID- 12588315 TI - Renal allograft arteriovenous fistula and large pseudoaneurysm. AB - The patient was a 51-year-old female. Post-biopsy arteriovenous fistula (AVF) and pseudoaneurysm in a renal allograft were diagnosed 5 yr and 4 months after she received a renal transplantation. Four years after the diagnosis, interventional treatment for the AVF and pseudoaneurysm was performed because of a high risk of pseudoaneurysm rupture. Although the longitudinal diameter of the pseudoaneurysm was more than 5 cm, this AVF and pseudoaneurysm were treated successfully by a percutaneous transluminal embolization, and renal function has remained stable after embolization. A selective interventional procedure proved effective for the large pseudoaneurysm in the renal allograft. Therefore, when a transcutaneous needle biopsy of the renal allograft is performed, although there are no apparent symptoms or signs of vascular complications during the clinical course, periodical examinations such as echo-Doppler imaging should be made on the allograft. PMID- 12588316 TI - Predominant factors associated with bone loss in liver transplant patients - after prolonged post-transplantation period. AB - INTRODUCTION: Osteoporosis is a major cause of morbidity in liver transplant recipients and is associated with multiple factors. OBJECTIVES: To evaluate bone mineral density (BMD), bone turnover and calcium-regulating hormones in 29 patients (17 men, 12 women) 2-12 yrs following liver transplantation for non alcoholic liver diseases. RESULTS: Fifteen patients (52%) were on immunosuppressive treatment with tacrolimus and 14 (48%) with cyclosporine. Eleven patients (38%) were currently on prednisone, 18 patients (62%) had stopped glucocorticoid treatment 6 months to 11 yrs prior to the study. Nineteen patients (65.5%) had decreased BMD according to WHO criteria, 17 (58.2%) at the femoral neck, 13 (44.8%) at the lumbar spine. Nineteen patients (65.5%) had a subnormal (<15 ng/mL) serum level of 25 (OH) D3. These patients had significantly lower BMD at the femoral neck (p = 0.02). Femoral neck BMD negatively correlated with serum parathyroid hormone level (p = 0.06, r = -0.35), length of the post transplantation period (p = 0.025, r = -0.416) and duration of glucocorticoid treatment (p = 0.029, r = -0.406), regardless of its cumulative dose. Symptomatic fractures were less frequent in tacrolimus treated patients than in cyclosporine users (p = 0.03). CONCLUSIONS: Decreased BMD is frequent following liver transplantation and is affected by vitamin D deficiency, cyclosporine use, and the duration of glucocorticoid therapy, but not by its cumulative dose. Achievement and maintenance of optimal vitamin D status and shortening of glucocorticoid treatment period may have a favorable effect on bone preservation. PMID- 12588317 TI - Failure of cyclosporin A to induce transforming growth factor beta (TGF-beta) synthesis in activated peripheral blood lymphocytes. AB - Induction of transforming growth factor beta (TGF-beta) by the immunosuppressive drug cyclosporin A (CsA) in activated lymphocytes has been claimed to add to the renal pro-fibrotic effects of CsA. The aim of this study was to evaluate CsA mediated TGF-beta induction in a larger number of lymphocyte preparations from different donors. Peripheral blood lymphocytes (PBL) were obtained from healthy blood donors. The cells were stimulated with phytohemagglutinin E (PHA) and phorbol ester (tetradecanoyl phorbol acetate, TPA) in the presence or absence of CsA. TGF-beta, interleukin-2 (IL-2) and cyclooxygenase-2 (Cox-2) mRNA were detected by Northern blot analysis or by real time reverse transcriptase polymerase chain reaction (RT-PCR). TGF-beta and IL-2 protein were determined in the cellular supernatants by enzyme-linked immunosorbent assay. TGF-beta mRNA and protein were up-regulated when the cells were stimulated with PHA/TPA. Cyclosporin A at high concentrations (500 ng/mL) caused a transient increase in TGF-beta mRNA which was significant after 2 h. CsA did not induce sustained TGF beta protein expression (24-72 h) in any of the preparations (n = 14), whereas the up-regulation of IL-2 mRNA and protein was prevented by CsA in the same preparations. Furthermore, up-regulation of Cox-2 mRNA was inhibited by CsA. Taken together, there was no evidence for TGF-beta as a clinically relevant mediator being induced by CsA in activated peripheral blood T-lymphocytes. PMID- 12588318 TI - Urodynamic testing predicts long-term urological complications following simultaneous pancreas-kidney transplantation. AB - INTRODUCTION: Combined pancreas-kidney transplantation is the treatment of choice for patients with type I diabetes mellitus associated with chronic renal failure. The introduction of the bladder drainage technique constituted a marked improvement of the surgical technique with a reduction of life-threatening complications. However, drainage of pancreatic secretions via the urinary bladder causes urological complications leading, in some cases, to cystoenteric conversion. We retrospectively analysed whether pre-operative urodynamic findings may predict the subsequent development of urological complications and influence the choice of exocrine secretion drainage. PATIENTS AND METHODS: From 1987 to 1997, 39 bladder-drained simultaneous pancreas-kidney transplantations were performed in 16 men and 23 women with a mean age of 38.5 yr. All patients underwent a complete urological assessment prior to surgery, including medical history, physical examination, urethrocystography and urodynamic assessment. RESULTS: Twenty-eight patients are alive with a mean follow-up of 62 +/- 8 months. In 60% of cases, both kidney and pancreas remain functional. Seven patients experienced recurrent lower urinary tract infections. Six patients suffered from chemical urethritis (four men and two women) and six suffered from recurrent haematuria (blood transfusions were required in two patients). One patient had incrusted stones at the site of duodenal staples. Urological complications were mostly observed in the 22 patients (79%) with abnormal urodynamic characteristics (Relative risk: 5.1). Intravenous Somatostatin failed to definitively cure these complications in most cases. Seven patients (17%) (five with urethritis, two with haematuria) required cystoenteric conversion. Two patients developed post-operative ileal fistula, one cutaneous and one into the bladder. All urinary symptoms resolved in these seven patients. CONCLUSION: The frequency of specific urinary complications is high (28%) in bladder-drained simultaneous pancreas-kidney transplantation patients. These complications are statistically more frequent in the case of an abnormal pre-transplant urodynamic assessment. PMID- 12588319 TI - Acute pancreatitis following kidney transplantation - role of viral infections. AB - Acute pancreatitis following renal transplantation is an unusual complication that carries a high mortality. Over the last 10 yr, five of 185 patients at our center developed acute pancreatitis. All had live related donors and were on conventional triple drug immunosuppression. Pancreatitis was classified according to the computed tomography scan based on Atlanta Classification. All five patients who developed acute pancreatitis had evidence of symptomatic or serologically active viral infection (chicken pox in two, cytomegalovirus infection in two, hepatitis E virus in one) and no patient without viral infection developed pancreatitis. Overall, 45 patients developed symptomatic or serologically active viral infection. There was a significant association between viral infection and pancreatitis (chi-square test, p < 0.001). Three patients with severe acute pancreatitis died while both patients with mild pancreatitis survived. An active search for viral infections should be made in all patients with acute pancreatitis. Specific antiviral measures may help reduce the mortality of acute pancreatitis in these patients. Consideration must be given to varicella immunization in patients with renal failure. PMID- 12588320 TI - Technique of bile duct reconstruction and management of biliary complications in right lobe living donor liver transplantation. AB - From December 1999 to January 2002, 50 right lobe living donor liver transplantations were performed. The donor operations included an intraoperative cholangiography to elicit variations in bile duct anatomy. The biliary reconstruction was done whenever possible as an end-to-end microanastomosis of the donor right hepatic duct with the recipient's bile duct. As a result of the early segmental branching of the donor biliary tree, two segment bile ducts had to be anastomosed in 20 patients and three segment bile ducts in three patients. In 12 patients, a Roux-en-Y hepaticojejunostomy was performed. All anastomoses were drained externally. We observed two leakages at the resection surface which could be treated successfully by an external drainage. Six leaks occurred at the site of end-to-end biliary anastomoses. Twice the problem could be conservatively solved placing a stent percutaneously. In two patients a hepaticojejunostomy was performed after a bile duct necrosis. In two patients with an anastomotic leak, occurring 3 d, respectively, 3 month after the original transplantation, the bile duct could be directly reconstructed over a T-tube. Two anastomotic stenoses were observed, one in combination with a leak treated by percutaneous stent implantation and the second, 3 month after transplantation which was treated surgically. Biliary reconstruction after living donor liver transplantation requires microsurgical techniques and can be performed as a direct end-to-end anastomosis in most cases. Biliary complications were treated by percutaneous drainage or surgical revision in all cases. PMID- 12588321 TI - Five-year results of renal transplantation on immunosuppressive triple therapy with mycophenolate mofetil. AB - Mycophenolate mofetil (MMF) combined with cyclosporine and prednisolone significantly lowers acute rejection frequency in the early post-renal transplantation phase. To date only registry data with very high transplant numbers have shown that MMF significantly influences long-term outcome. A comparative retrospective analysis of the 5-yr results with MMF in a single transplant center was thus undertaken vs. other standard immunosuppressive regimens. The results of 1579 renal transplantations were grouped by treatment modality, subjected to Kaplan-Meier analysis, and compared using the log rank test. Both the total population and subgroups showed a non-significant trend towards better graft survival with MMF, evident at 2 yr and persisting for 5 yr. Extrapolation indicates that on combination therapy with MMF vs. azathioprine, approximately 10% more patients will be alive at 10 yr with a functional graft. PMID- 12588322 TI - Haemodynamic monitoring and liver function evaluation by pulsion cold system Z 201 (PCS) during orthotopic liver transplantation. AB - Pulsion cold system (PCS, COLD) is a haemodynamic monitoring system that allows measurement of cardiac output (CO), partial blood volumes, lung water, and liver function. The aim of the study was to evaluate this monitoring system during human orthotopic liver transplantation (OLT) for the following: (a) to determine agreement between CO measurements via pulmonary artery thermodilution (CO TDpa), and aortic transpulmonary thermodilution (CO TDa); (b) to compare the preload dates obtained with the COLD with central venous pressure (CVP) and pulmonary capillary wedge (PCWP); and (c) to assess the use of the plasma disappearance rate (PDR) of indocyanine green (ICG) as a measure of graft function. Fifteen consecutive patients undergoing OLT were studied. Each patient received a pulmonary artery catheter and a 5F aortic catheter with an integrated thermistor. The thermistor of the aortic catheter were connected to one computer system (COLD Z201, Pulsion Medical Systems, Munich, Germany). Haemodynamic data were registered an all the phases of OLT. PDR was measured during surgery in 12 patients. Correlations between PDR and the other markers of graft function (transaminases, protrombine time, and bile production) were sought. The correlation coefficient between CO TDa (COLD) and CO TDpa was r = 0.766 (p < 0.001), and an additional analysis according to Bland-Altman was also performed. There was a better correlation between the cardiac index (determined by two monitoring systems) and the volume measurements than the correlation observed with pressure preload parameters. The best correlations were found between the cardiac index in the femoral artery and intrathoracic blood volume index (ITBVI) and pulmonary blood volume index (PBVI) (r = 0.79 and r = 0.72, respectively; p < 0.01). PDR measured in the group patients with bad early graft function were lower (13.6 +/- 2.7) than those in the group with a good graft function (21.6 +/- 9) (p < 0.05). The degree of discrepancy between femoral and pulmonary thermodilution cardiac output measures is very wide during OLT so as to make the techniques using the COLD machine clinically useless. On the other hand, the volumes measured by COLD, specially ITBVI and PBVI, are more useful to asses the pre-load than pressure measurements. In OLT, the PDR measured within the first few hours after liver reperfusion may become a useful tool for early diagnosis of primary graft dysfunction (PDF). PMID- 12588324 TI - The changes in serum leptin, body fat mass and insulin resistance after renal transplantation. AB - BACKGROUND: In this prospective-controlled study, we aimed to investigate the effect of changes in insulin resistance and anthropometrical parameters on serum leptin levels (SLL) after renal transplantation (Tx). PATIENTS AND METHODS: Thirty-four patients (M/F: 19/15, mean age: 29 +/- 9 yr) and 30 age and sex matched healthy controls (C) were included. Body weight, subscapular, suprailiac, periumbilical, biceps and triceps skinfold thicknesses, neck, wrist, hip and waist circumferences, as well as body mass index and body fat mass were measured as anthropometrical parameters. In order to measure the serum glucose, insulin and SLL, blood samples were obtained before and 1 wk, 1 and 6 months after Tx. Homeostasis Model Assessment (HOMA) values were calculated as an index of insulin resistance. RESULTS: Serum leptin levels (SLL) of the patients at pre-Tx were significantly higher than C (21.5 +/- 3.5 vs. 7.8 +/- 0.9 ng/mL, p = 0.002) and decreased at first week after Tx (from 21.5 +/- 3.5 to 8.4 +/- 1.5 ng/mL, p < 0.001). Thereafter, it gradually increased to 12.8 +/- 2.1 ng/mL in the first month and to 14.4 +/- 2.1 ng/mL in the sixth month after Tx. Serum leptin levels at sixth month were significantly higher than C (p = 0.005). Serum insulin and HOMA values changed similar to SLL after Tx. Correlations between SLL and HOMA persisted during the study period [pre-Tx (r: 0.40) and at first (r: 0.38) and sixth (r: 0.47) months]. In linear regression analysis, HOMA and fat mass were found as independent variables for predicting SLL at the sixth month after Tx. CONCLUSION: Serum leptin levels dramatically decreased immediately after Tx and significantly correlated with serum insulin levels and HOMA during the entire study. Increase in SLL at sixth months was probably because of increase in fat mass, insulin resistance and steroid use in renal transplant recipients. PMID- 12588323 TI - The apo A-I gene promoter region polymorphism determines the severity of hyperlipidemia after heart transplantation. AB - BACKGROUND: To study whether the Apolipoprotein A-I (apo A-I) promoter region gene polymorphism produces changes in the lipid profile of heart transplant recipients. METHODS: One hundred and three heart transplant recipients (93 men and 10 women, with a mean age of 47 +/- 13 yr) receiving triple immunosuppressive therapy were submitted to a genetic study of the apo A-I gene promoter region. Anthropometric and analytical data, including lipid profile, arterial blood pressure, were collected prior to transplantation and 3, 6, 12, and 24 months after transplantation. RESULTS: Sixty-three subjects had the GG genotype and 40 the GA genotype. Carriers of the GA genotype had higher triglyceride levels at 6 months and 2 yr (2.50 +/- 1.20 versus 1.93 +/- 0.98 mmol/L and 2.46 +/- 1.58 versus 1.60 +/- 0.68 mmol/L, respectively, p < 0.001), and a greater rise in LDL cholesterol at 1 yr than the GG subjects (4.57 +/- 1.16 versus 4.16 +/- 1.18 mmol/L, p < 0.05). Multiple regression analyses showed that genetic variants at the apo A-I promoter region are responsible for 11% of the variability in triglyceride levels at 6 months (p = 0.005). CONCLUSIONS: The GA genotype of the apo A-I promoter region produces a greater rise in plasma triglyceride and LDL cholesterol levels in heart transplant patients. PMID- 12588325 TI - Individualized T cell monitored administration of ATG versus OKT3 in steroid resistant kidney graft rejection. AB - Acute steroid-resistant rejection episodes are recommended to be treated with set doses of anti-thymocyte globulin (ATG) or anti-CD3 monoclonal antibody (OKT3). Individualized T cell monitoring has been proposed as a tool for dose finding. A randomized study comparing the efficacy and safety of ATG (n = 27) with OKT3 (n = 28) in the treatment of biopsy verified acute steroid-resistant rejection (ASRR) when both drugs were administered on the basis of daily individualized T cell measurements. A drop to below 50 cells/mm3 CD2+ T cells was considered adequate and used to guide the dose of ATG/OKT3. Demographic, clinical and histopathological severities of rejections were equal in the two groups. During the 10 days of T cell monitoring and antibody treatment, 13 patients were in need of dialysis (ATG = 7/OKT3 = 6). Two grafts did not respond to antibody treatment and were lost due to rejection (ATG = 1/OKT3 = 1). There were 26 biopsy verified re-rejections (ATG = 12/OKT3 = 14) within the first 3 months following antibody treatment. Mean serum creatinine (micromol/L) was similar in the two groups (ATG/OKT3: before rejection 157 +/- 72/151 +/- 88, at start of antibody treatment 308 +/- 125/330 +/- 94, end of antibody treatment 254 +/- 122/246 +/- 144 and at follow-up after a mean of 32 months 166 +/- 55 (n = 24)/164 +/- 57(n = 23)). To keep the T cell count below 50 cells/mm3, average dose ATG given was 354 +/- 151 mg (2.3 administrations, range 1-4) and average OKT3 was 32.5 +/- 6.8 mg in 10 doses. In conclusion, individualized T cell monitored administration of ATG and OKT3 is safe and seems as effective as a standard set dose in treatment of ASRR. Tailoring the dose for each individual patient lowers the cost. PMID- 12588326 TI - Long-term outcome of kidney transplantation in patients with hepatitis B or C positive. PMID- 12588327 TI - Carbohydrate-protein conjugate vaccines. AB - Various pathogenic bacteria have coats of polysaccharide, many with repeating epitopes. Though polysaccharide vaccines have been available for some time, they induce mainly IgM production, and are only moderately protective in adults and ineffective in young children. It was originally shown in 1931 that the immunogenicity of polysaccharides could be enhanced by conjugating to a protein. The last two decades have witnessed the production and clinical testing of polysaccharide-protein conjugates specific for at least four different bacteria which normally cause considerable mortality and morbidity, especially in young children. In some cases, immunizing children from 4 months of age, with a booster early in the second year, has resulted in remarkably high success rates in protecting them from disease. For one pathogen, Haemophilus influenza type b, the success rate has been sufficiently high (> 95%) to suggest that this disease might, in time, be globally controlled in this way. The results of immunization with conjugate vaccines to Streptococcus pneumoniae, Neisseria meningiditis and Salmonella typhi are also very encouraging. More conjugate preparations are under development. PMID- 12588328 TI - High-throughput epidemiologic typing in clinical microbiology. AB - Mapping, and ultimately preventing, the dissemination of infectious agents is an important topic in public health. Newly developed molecular-microbiological methods have contributed significantly to recent advances in the efficient tracking of the nosocomial and environmental spread of microbial pathogens. Not only has the application of novel technologies led to improved understanding of microbial epidemiology, but the concepts of population structure and dynamics of many of the medically significant microorganisms have advanced significantly also. Currently, genetic identification of microbes is also within the reach of clinical microbiology laboratory professionals including those without specialized technology research interests. This review summarizes the possibilities for high-throughput molecular-microbiological typing in adequately equipped medical microbiology laboratories from both clinical and fundamental research perspectives. First, the development and application of methods for large-scale comparative typing of serially isolated microbial strains are discussed. The outcome of studies employing these methods allows for long-term epidemiologic surveillance of infectious diseases. Second, recent methods enable an almost nucleotide-by-nucleotide genetic comparison of smaller numbers of strains, thereby facilitating the identification of the genetic basis of, for instance, medically relevant microbiological traits. Whereas the first approach provides insights into the dynamic spread of infectious agents, the second provides insights into intragenomic dynamics and genetic functionality. The current state of technology is summarized, and future perspectives are sketched. PMID- 12588329 TI - Infecton: a 99mTc-ciprofloxacin radiopharmaceutical for the detection of bone infection. AB - OBJECTIVE: To evaluate Infecton scintigraphy, with technetium-99m-radiolabeled ciprofloxacin, as a means to detect bone infection, in comparison with other conventional scintigraphic and radiologic methods. METHODS: Forty-five patients with known or suspected bone infection underwent 50 scans with Infecton. Almost all were also subjected to a three-phase 99mTc-methylene diphosphonate bone scan and most of them to a 99mTc-human polyclonal immunoglobulin scan as well as to a gallium-67-citrate scan, plus computerized tomography or magnetic resonance imaging or both. Clinical laboratory criteria for the presence of osteomyelitis were based on the definitions of the Centers for Disease Control and Prevention. RESULTS: Staphylococcus aureus and Pseudomonas aeruginosa were the most frequently isolated pathogens. Based on the CDC clinical laboratory criteria as well as on conventional scan results, Infecton was characterized in 35 studies as 'true positive', in eight as 'true negative', in two as 'false positive', in one as 'false negative', and in four as 'indeterminate'. The sensitivity and specificity of Infecton scintigraphy were found to be 97.2% and 80%, respectively, with positive and negative predictive values of 94.6% and 88.9%. CONCLUSIONS: It is concluded that Infecton is a very sensitive and quite specific marker of bone infection, but care must be taken in cases of excessive new bone formation and primary bone tumors, where false-positive results may be obtained. PMID- 12588330 TI - Dientamoeba fragilis, a neglected cause of diarrhea, successfully treated with secnidazole. AB - OBJECTIVE: To evaluate the pathogenicity of Dientamoeba fragilis by comparing it with Giardia lamblia and to investigate the effect of a single dose of secnidazole in dientamoebiasis. METHODS: Stool samples of 400 patients, admitted to the Department of Parasitology, Celal Bayar University, were examined by direct wet mount, formalin-ethyl acetate concentration and trichrome staining methods on three consecutive days. All cases positive for D. fragilis were treated with a single dose of secnidazole, 30 mg/kg for children, and 2 g for adults. On the seventh and fourteenth days, at the end of the treatment, stool samples were examined by the same methods, and clinical symptoms were again evaluated. RESULTS: D. fragilis and G. lamblia were detected in 35 (8.8%) and 34 (8.5%) cases, respectively. The most frequent symptoms were found to be abdominal pain and diarrhea in both infections. D. fragilis was eradicated in 34 (97.1%) patients with a single dose of secnidazole, but a second dose was necessary in one patient. Clinical symptoms disappeared in 27 (77.1%) and decreased in eight (22.9%) cases, after eradication. CONCLUSIONS: These data suggested that D. fragilis is as prevalent and pathogenic as G. lamblia, and secnidazole seems to be highly effective in achieving parasitologic and clinical cure. To our knowledge, this is the first report of secnidazole being used in the treatment of dientamoebiasis. PMID- 12588331 TI - Reliability of the ica, aap and atlE genes in the discrimination between invasive, colonizing and contaminant Staphylococcus epidermidis isolates in the diagnosis of catheter-related infections. AB - OBJECTIVES: To evaluate the usefulness of detecting two genes involved in biofilm formation (icaA and aap) and one gene involved in initial adhesion (atlE) for discrimination between contaminant, colonizing and invasive Staphylococcus epidermidis isolates involved in catheter-related infections. PATIENTS: The first group contained 29 isolates that were isolated from the skin of healthy volunteers (contaminant isolates). The second group contained 16 isolates recovered from catheters (>1000 CFUs on quantitative catheter culture) from asymptomatic patients without bacteremia. These isolates were considered to be colonizing isolates. The third group contained 34 isolates grown in >or=2 different blood cultures from patients with a systemic inflammatory response. These isolates were considered to be invasive isolates. RESULTS: The prevalence of atlE did not differ between the three groups. The icaA and aap genes were significantly more prevalent in colonizing isolates (88% aap; 88% icaA) than in invasive isolates (68% aap, P = 0.179; 59% icaA, P = 0.055) and than in skin isolates (52% aap, P = 0.02; 38% icaA, P = 0.002). CONCLUSIONS: The high prevalence of aap and icaA in skin isolates and their higher prevalence in colonizing than in invasive isolates led to a low specificity when these genes were used to differentiate between contamination, colonization and invasive infection. We conclude that, although the prevalence of these genes differs in the three groups, their presence cannot be used for clinical decision-making. PMID- 12588333 TI - Clinical microbiological case: acute osteomyelitis in a previously healthy child. PMID- 12588332 TI - A new method for normalized interpretation of antimicrobial resistance from disk test results for comparative purposes. AB - OBJECTIVE: To evaluate a calibration method for disk diffusion antibiotic susceptibility tests, using zone diameter values generated in the individual laboratory as the internal calibrator for combinations of antibiotic and bacterial species. METHODS: The high-zone side of zone histogram distributions was first analyzed by moving averages to determine the peak position of the susceptible population. The accumulated percentages of isolates for the high zone diameter values were calculated and converted into probit values. The normal distribution of the ideal population of susceptible strains was then determined by using the least-squares method for probit values against zone diameters, and the ideal population was thereby defined, including mean and standard deviation. Zone diameter values were obtained from laboratories at the Karolinska Hospital (KS) and Vaxjo Hospital (VX), and from two laboratories (LabA, LabB) in Argentina. The method relies on well standardized disk tests, but is independent of differences in MIC limits and zone breakpoints, and does not require the use of reference strains. Resistance was tentatively set at below 3 SD from the calculated, ideal mean zone diameter of the susceptible population. RESULTS: The method, called normalized interpretation of antimicrobial resistance, was tested on results from the KS and VX clinical microbiology laboratories, using the disk diffusion method for antimicrobial susceptibility tests, and for two bacterial species, Staphylococcus aureus and Escherichia coli. In total, 114 217 test results were included for the clinical isolates, and 3582 test results for control strains. The methodology at KS and VX followed the standard of the Swedish Reference Group for Antibiotics (SRGA). Zone diameter histograms for control strains were first analyzed to validate the procedure, and a comparison of actual means with the calculated means showed a correlation coefficient of r = 0.998. Results for clinical isolates at the two laboratories showed an excellent agreement for 54 of 57 combinations of antibiotic and bacterial species between normalized interpretations and the interpretations given by the laboratories. There were difficulties with E. coli and mecillinam, and S. aureus and tetracycline and rifampicin. The method was also tested on results from two laboratories using the NCCLS standard, and preliminary results showed very good agreement with quality-controlled laboratory interpretations. CONCLUSIONS: The normalized resistance interpretation offers a new approach to comparative surveillance studies whereby the inhibition zone diameter results from disk tests in clinical laboratories can be used for calibration of the test. PMID- 12588334 TI - Extraction of Chlamydia pneumoniae DNA from vascular tissue for use in PCR: an evaluation of four procedures. AB - The objective of this study was to compare four procedures for Chlamydia pneumoniae DNA extraction from vascular tissue. The NucliSens Kit, the QIAamp tissue DNA MiniKit, buffer-saturated phenol and the Geneclean II Kit were evaluated, based on the yield of recovered DNA, using PCR to detect C. pneumoniae in vascular tissue. The QIAamp tissue procedure had the highest detection level (0.004 inclusion-forming units/sample). All methods, except NucliSens (70 min), had a short handling time (30-40 min). Costs varied from 0.5 to 3.2 Euro. PMID- 12588335 TI - Evidence for efflux pumps, other than PmrA, associated with fluoroquinolone resistance in Streptococcus pneumoniae. AB - Fluoroquinolone resistance in pneumococci is known to be associated with the efflux pump, PmrA. However, there may be other efflux systems that also cause drug resistance. Two types of mutants were studied. The efflux phenotype from mutants selected by sub-MIC levofloxacin or gemifloxacin was transformed into R6. These transformants did not show increased pmrA transcripts in Northern blots; insertional inactivation of pmrA in the transformants did not abolish the efflux phenotype. A second set of efflux phenotype mutants was selected in R6:cat by ethidium bromide but not by norfloxacin; accumulation of ethidium bromide in the one among these mutants studied was reduced in comparison to its parent. This evidence suggests that systems other than PmrA can contribute to efflux-mediated resistance in pneumococci. PMID- 12588336 TI - Human papillomavirus DNA in cervical lesions from Morocco and its implications for cancer control. AB - To determine the types of human papillomaviruses (HPV) in northern Morocco, information which is needed for the design and use of HPV vaccines, we have analysed 129 cervical biopsies from this region. In our study, 91 cases were HPV positive, 45 cases had HPV-16 DNA, and 20 cases had HPV-18 DNA. This distribution of virus type was similar in inflammatory cervical lesions and in invasive cervical carcinomas. In conclusion, the HPV type distribution in Morocco is similar to that in other African Mediterranean countries, where the proportion of HPV-18 cases is significantly higher than in Europe. Determination of virus-type distribution is essential for vaccination programs. PMID- 12588337 TI - Resistance to beta-lactams among blood isolates of Salmonella spp. in European hospitals: results from the SENTRY Antimicrobial Surveillance Program 1997-98. AB - The susceptibility to beta-lactams and the beta-lactamase content of 110 Salmonella spp. blood isolates collected during 1997-98 in 19 European centers participating in the SENTRY Surveillance Program were studied. Thirty-one isolates (28%) were resistant to penicillins, due to production of TEM-1 (27 isolates), OXA-1 (three isolates) or TEM-1 + OXA-1 (one isolate). All OXA-1 producers and 10 TEM-1-producing isolates were also resistant to penicillin clavulanic acid combinations. In the latter isolates, this phenotype was associated with increased production of TEM-1. Sixteen TEM-1-producing Salmonella Enteritidis isolates and one OXA-1-producing S. Typhimurium isolate were able to transfer beta-lactam resistance by conjugation. PMID- 12588338 TI - Prevalence of methicillin-resistant Staphylococcus aureus in eight African hospitals and Malta. AB - Methicillin-resistant Staphylococcus aureus (MRSA) poses a serious therapeutic problem worldwide, and its frequency in most African countries has not been reported. This study was aimed at determining the prevalence and antibiotic susceptibility patterns of MRSA in eight large hospitals (>500 beds) in Africa and Malta, from 1996 to 1997. Susceptibility to methicillin (oxacillin) and to other drugs was determined by E test (AB Biodisk, Solna, Sweden) on a total of 1440 clinical isolates of S. aureus. Methicillin resistance was detected in 213 (15%) of the 1440 isolates tested. The rate of MRSA was relatively high in Nigeria, Kenya, and Cameroon (21-30%), and below 10% in Tunisia, Malta, and Algeria. All MRSA isolates were sensitive to vancomycin, with MICs 60%) of MRSA strains were multiresistant. There is a need to maintain surveillance and control of MRSA infections in Africa. PMID- 12588340 TI - Cerebellar involvement in anaplastic large cell lymphoma. PMID- 12588341 TI - Chronic myelomonocytic leukaemia (CMML) presenting as a thyroid mass. PMID- 12588342 TI - A history of St Jude Children's Research Hospital. PMID- 12588343 TI - Guidelines on the diagnosis and management of the thrombotic microangiopathic haemolytic anaemias. PMID- 12588344 TI - Guidelines for the investigation and management of idiopathic thrombocytopenic purpura in adults, children and in pregnancy. PMID- 12588345 TI - Diagnosis of thrombotic thrombocytopenic purpura based on modulation by patient plasma of normal platelet adhesion under flow condition. AB - We have designed a simple test for the early diagnosis and treatment monitoring of thrombotic thrombocytopenic purpura (TTP). We examined plasma from 24 TTP patients and normal plasma using a cone and plate(let) analyser (CPA). Test plasma was mixed with citrated normal whole blood (group O) and subjected to flow at a shear rate of 1800/s. Mixing normal plasma (12.5, 25, 50 or 75 microl) with heterologous normal whole blood (final volume of 200 microl) resulted in a decrease of surface coverage (SC, maximally by 63%) and, to a lesser extent, of average size (AS, maximally by 37%) due to dilution of the blood sample. In contrast, mixing the same quantities of acute TTP plasma with normal blood yielded an increase in both SC (up to 125%) and AS (up to 130%). Increased SC and/or AS were detected in all 15 patients in acute phase and in three out of 14 patients in remission. Following repeated plasmapheresis, the enhanced platelet deposition in five patients with acute TTP returned to almost normal patterns. Mixing plasma from patients with other thrombocytopenic conditions in this way resulted in a decrease in both SC and AS, and did not differ from control subjects. In conclusion, the CPA is a simple and specific laboratory test that can be used for the diagnosis and monitoring of plasma exchange therapy in TTP. PMID- 12588346 TI - A peptide (P2) derived from the variable heavy chain of an anti-P-selectin monoclonal antibody (LYP20) inhibits leucocyte adhesion to thrombin-activated platelets and endothelial cells. AB - P-selectin, a member of the selectin family of adhesion molecules, is present in endothelial Weibel-Palade bodies and platelet alpha-granules, and is rapidly expressed on their surface upon activation, resulting in leucocyte adhesion. LYP20 is a functional monoclonal antibody previously generated in our laboratory that binds with high affinity and specificity directed against P-selectin. This binding is largely imparted by the specific sequence of amino acids present on the hypervariable portions of the IgG chains. We now show that a peptide derived from the heavy chain of mAb LYP20 dose dependently inhibits the adhesion of poly morphonuclear cells to resting and thrombin-activated endothelial cells (EC) and platelets. The scrambled form of this peptide, identical in amino acid composition to the authentic peptide but with altered sequence, was not inhibitory at corresponding concentrations. Binding studies revealed that this peptide also dose dependently bound to both resting and thrombin-activated EC and platelets. Our results may prove useful for the development of new therapeutic inhibitors to modulate leucocyte interactions in inflammatory disorders. PMID- 12588347 TI - Inhibition of factor VIIa generation and prothrombin activation by treatment with enoxaparin in patients with unstable angina. AB - Factor VIIa (FVIIa) and thrombin generation occur in patients suffering an acute coronary event. We studied the effect of treatment with enoxaparin on FVIIa and prothrombin activation in patients with unstable angina. Anti-Xa activity, FVIIa, FVII coagulant activity (FVII:C) and FVII antigen (FVII:Ag), free tissue factor pathway inhibitor (TFPI), and prothrombin fragments 1 + 2 (F1+2) were measured in patients' plasma, over a 24-h treatment period with enoxaparin. All 14 patients recruited in the study (mean age 68 years) were treated with a subcutaneous injection of enoxaparin, 1 mg/kg twice daily. Blood was drawn just before, and at different time intervals after, the first injection. Before enoxaparin administration, the levels of FVIIa (4.02 +/- 0.8 ng/ml) and F1+2 (2.68 +/- 0.2 nmol/l) were significantly increased as compared with control subjects (2.3 +/- 0.3 ng/ml and 0.9 +/- 0.1 nmol/l respectively, P < 0.05). Free TFPI, FVII:C and FVII:Ag were within normal ranges. One hour after the first injection of enoxaparin, FVIIa and F1+2 levels decreased by 65% and 50%, respectively, and no significant fluctuations were noted throughout the observation period. The concentrations of FVII:C and FVII:Ag were not modified as compared with baseline values. After each injection, the peak concentrations of free TFPI and anti-Xa activity were observed at 2 and 4 h respectively. The kinetics of FVIIa and F1+2 inhibition did not follow those of anti-Xa activity and TFPI release. PMID- 12588348 TI - Factor VII/VIIa: a new antigen in the anti-phospholipid antibody syndrome. AB - We investigated antibodies to factor VII/VIIa (FVII/VIIa) and five other common target antigens in 33 patients with a history of anti-phospholipid syndrome (APS) and 50 healthy controls using an enzyme-linked immunosorbent assay (ELISA) technique. We found that antibody to FVII/VIIa, a previously unrecognized and common antigen in APS, was present in 67% of patients. Frequencies of antibodies to other target antigens were: anti-beta-2 glycoprotein 1 (anti-beta 2GP1), 88%; anti-cardiolipin (anti-CL), 76%; anti-phosphatidylethanolamine (anti-PE), 67%; anti-phosphatidylserine (anti-PS), 64%; and anti-phosphatidylcholine (anti-PC), 59%. Most patients had antibodies against multiple antigens, but a few were positive for only anti-beta 2GP1 (12%) or anti-CL (3%). Positivity for anti FVII/VIIa was significantly associated with positivity for anti-PE, anti-PS and/or anti-PC (P < 0.05) but not anti-beta 2GP1. When frequencies of immunoglobulin G (IgG) versus immunoglobulin M (IgM) antibodies were compared, anti-beta 2GP1 IgG correlated with the lupus anticoagulant (P < 0.05) and was significantly more prevalent than IgM, but the reverse was seen for all other antigens. In arterial thrombosis, IgM was more prevalent for all antigens, and was significantly associated with FVII/VIIa, PE and PS, whereas in venous thrombosis, IgG was frequently prevalent, especially in association with FVII/VIIa, beta 2GP1 and CL. In summary, FVII/VIIa is a new and common antigen in APS. Anti-FVII/VIIa is often associated with anti-PE, anti-PS and anti-PC. The IgM class is more frequently associated with arterial thrombosis and the IgG class with venous thrombosis. PMID- 12588349 TI - Two novel mutations, Q1053H and C1060R, located in the D3 domain of von Willebrand factor, are responsible for decreased FVIII-binding capacity. AB - In type 2N von Willebrand disease (VWD), von Willebrand factor (VWF) is characterized by a markedly decreased affinity for Factor VIII (FVIII), and the mutations responsible are essentially located in the D' domain of VWF. We report the identification, in seven unrelated French families, of two novel type 2N VWD mutations, Q1053H and C1060R (Gln290His and Cys297Arg in mature VWF sequence), in exon 24 of the VWF gene. These missense mutations have been identified in the heterozygous, homozygous or hemizygous states. Using site-directed mutagenesis and transient expression in COS-7 cells, we showed that both mutations, although located in the D3 domain of VWF, outside the tryptic fragment containing the FVIII domain, dramatically decrease the binding of VWF to FVIII. In contrast, the R924Q substitution, which was identified in a patient who was heterozygous for C1060R, was shown to be a polymorphism. PMID- 12588350 TI - Effect of clopidogrel administration to healthy volunteers on platelet phosphorylation events triggered by ADP. AB - The action of clopidogrel on platelet receptors was analysed using platelets obtained from 11 healthy volunteers given 75 mg of clopidogrel daily for 8 d. Samples of blood were taken before treatment and after 8 d of medication. Determination of 2-methylthioadenosine diphosphate trisodium (2MesADP)-induced platelet aggregation, serine/threonine and tyrosine phosphorylations were performed in the absence or presence of the P2Y1-receptor-specific antagonist: adenosine 3'-phosphate 5'-phosphate (A3P5P) or the strong inhibitor of GPIIb/IIIa activation: SR121566. MAJOR CONCLUSIONS: 1). Serine and threonine phosphorylations of the myosin light chain (P20) and pleckstrin (P47) do not behave similarly, although they are both recognized as the result of phospholipase C pathway stimulation triggered by the P2Y1 receptor. P47 is strongly affected by the A3P5P, and this appears to be highly dependent on P2Y12. However, P20 phosphorylation occurs in the presence of A3P5P, suggesting that the P2Y12 receptor signal contributes to P20 phosphorylation mediated by a calcium independent pathway. The results suggest that P2Y1 and P2Y12 receptors interact to modulate the phosphorylation of P20 and P47. 2). The inside-out signalling dependent on both P2Y12 and P2Y1 is necessary for GPIIb/IIIa activation. 3). Clopidogrel and SR121566 inhibited the increase in tyrosine phosphorylation induced by 2MesADP and concomitantly inhibited platelet aggregation, indicating that most of the phosphorylations are GPIIb/IIIa dependent. However, neither clopidogrel nor SR121566 inhibited the first wave of 80 kDa substrate (cortactin) which is involved in the reorganization of the cytoskeleton necessary for shape change and which appeared to be essentially P2Y1 dependent. PMID- 12588351 TI - A new mutation, S1285F, within the A1 loop of von Willebrand factor induces a conformational change in A1 loop with abnormal binding to platelet GPIb and botrocetin causing type 2M von Willebrand disease. AB - We report the identification of a new mutation in exon 28 of the von Willebrand factor (VWF) gene in two related patients with type 2M von Willebrand disease (VWD). The molecular abnormality changes the Ser 1285 to Phe within the A1 loop of VWF. The S1285F mutation was reproduced by site-directed mutagenesis on the full-length VWF cDNA. The mutated recombinant VWF (rVWF), F1285rVWF, and the hybrid, S/F1285rVWF, were expressed in COS-7 cells. F1285rVWF exhibited a slight decrease of high-molecular-weight multimers and markedly reduced ristocetin- or botrocetin-induced binding of VWF to platelets in association with a decreased binding to botrocetin. The hybrid S/F1285rVWF showed a normal multimeric profile and bound to platelets in a similar way to the patients' plasma VWF, in the presence of ristocetin or botrocetin. Thus, the new S1285F mutation within the A1 loop was responsible for the type 2M VWD observed in these patients, and was involved in the binding of VWF to botrocetin and to platelet glycoprotein Ib (GPIb). Three anti-VWF monoclonal antibodies, with conformational epitopes within the A1 loop but distinct GPIb binding inhibitory properties, showed a different interaction with F1285rVWF. These results indicate that the S1285F substitution alters the folding of the A1 loop and prevents the correct exposure of the VWF binding sites to botrocetin and GPIb. PMID- 12588352 TI - Glycoprotein Ib polymorphisms influence platelet plug formation under high shear rates. AB - Platelet polymorphisms (Kozak, VNTR and HPA-2) within glycoprotein (GP)Ib alpha may be associated with an increased risk of arterial thrombosis. However, the functional role of these polymorphisms has not been clarified. Their influence on platelet plug formation under high shear rates was, therefore, examined in 233 healthy individuals. Collagen-adrenaline-induced closure time was shorter in carriers of the C/D versus C/C VNTR allele and in homozygotes with the (-5)T/T versus (-5)C/T Kozak genotype as determined by novel polymerase chain reaction methods. The HPA-2 genotype had no effects, and the density of GPIb alpha molecules was not influenced by GPIb alpha genotypes. PMID- 12588353 TI - Molecular pathology of haemophilia B in Turkish patients: identification of a large deletion and 33 independent point mutations. AB - Heterogeneous mutations in the coagulation factor IX (FIX) gene result in a bleeding tendency known as haemophilia B. The haemophilia B mutation database has a total of 2353 patient entries, including 10 of the estimated 1000 Turkish patients. In this study, a more comprehensive analysis of the molecular pathology of haemophilia B in Turkey revealed one large deletion and 33 point mutations in the FIX gene of 34 unrelated patients. Haplotype analysis using six polymorphic sites showed that the mutations identified in a total of 45 patients occurred on 13 different haplotypes and that each mutation was family specific. PMID- 12588354 TI - Burkitt's and Burkitt-like lymphoma in children and adolescents: a review of the Children's Cancer Group experience. AB - Historically, the survival of children and adolescents with Burkitt's and Burkitt like lymphoma had been poor. Recently, short and intensive chemotherapy appears to have improved disease outcome. We therefore reviewed the results of four successive Children's Cancer Group trials conducted on 470 children with disseminated Burkitt's and Burkitt-like lymphoma. Of the patients studied, the median age was 8 years (0-21 years), the male:female ratio was 4:1, 58% had lactate dehydrogenase (LDH) > or = 500 IU/l, 23% had M2 or M3 bone marrow (BM), and 12% demonstrated central nervous system involvement. In a multivariate analysis, the 4-year event-free survival (EFS) in patients > or = 15-years-old compared with < 15-year-old was 34 +/- 7 versus 59 +/- 2% (P < 0.05), the 4-year EFS of M2/M3 compared with M1 BM was 38 +/- 5 versus 63 +/- 3% (P < 0.001), and the 4-year EFS with LDH > or = 500 IU/l compared with LDH < 500 IU/l was 49 +/- 3 versus 71 +/- 4% (P < 0.001). Furthermore, patients treated on the most recent protocol, which was short and more intensive, had a significantly improved survival compared with those on previous trials (4-year EFS 80 +/- 6 versus 54 +/ 2%, P < 0.001). In summary, the outcome for childhood Burkitt's and Burkitt-like lymphoma has recently improved with the use of short and intensive B-cell non Hodgkin's lymphoma-directed therapy. PMID- 12588355 TI - Leucocyte-specific protein (LSP1) in malignant lymphoma and Hodgkin's disease. AB - Biopsies from 319 haematopoietic neoplasms were immunostained for intracellular leucocyte-specific protein 1 (LSP1) to assess its distribution and to compare its diagnostic value with that of CD45 (leucocyte common antigen: LCA). Most small B cell neoplasms expressed LSP1, but one third of diffuse large B-cell lymphomas (DLBCL) were LSP1 negative. Among the cases with DLBCL (76 samples) tested for both LSP1 and CD45, one fifth expressed only CD45, but five samples were LSP1 positive and negative for CD45. The latter pattern was also seen in four of nine myelomas. Five out of 14 T-lymphoblastic lymphomas co-expressed LSP1 and CD45, and three cases were LSP1 negative and CD45-positive. Most peripheral T-cell lymphomas co-expressed LSP1 and CD45. All anaplastic lymphoma kinase (ALK) negative lymphomas of anaplastic large cell morphology (T and null phenotype) expressed LSP1 although the percentage of LSP1-positive tumour cells was variable, however, only seven out of 30 cases with ALK-positive lymphoma were LSP1 positive. LSP1 was expressed on Reed-Sternberg cells in 60 out of 66 cases with classic Hodgkin's disease but neoplastic cells were usually negative in lymphocyte predominant Hodgkin's. This study confirms the wide expression of LSP1 within haematopoietic neoplasms and its diagnostic value for a minority of lymphoid tumours that have lost CD45 expression. Furthermore, the strong expression of LSP1 in classic Hodgkin's disease, contrasting with its heterogeneous expression in ALK-negative anaplastic lymphomas, may help to distinguish the latter lymphomas from patients with tumour cell-rich Hodgkin's disease. PMID- 12588356 TI - A pilot study of antithymocyte globulin (ATG) in the treatment of patients with 'low-risk' myelodysplasia. AB - We report 30 'low-risk' patients with myelodysplasia (MDS) (defined as < 10% bone marrow blasts) who were treated with antithymocyte globulin (ATG). In total, 20 patients were evaluable at the study end-point (response to treatment at 6 months). The diagnosis in these 20 patients was refractory anaemia (RA) in 13, RA with excess blasts in four, and RA with ringed sideroblasts in three. Median age was 54.5 years (range, 31-73 years). There were two cases of secondary MDS. The bone marrow was hypocellular in eight cases and cytogenetics were abnormal in four cases. All patients received lymphoglobuline (horse ATG; Sangstat, France) at a dose of 1.5 vials/10 kg/day for 5 d. The treatment was well tolerated. Three patients in the study died (disease progression, invasive aspergillosis and lung carcinoma respectively); 10 out of 20 evaluable patients (50%) responded to treatment and became transfusion independent; eight out of 13 (62%) patients with RA responded. The median duration of response was 15.5 months (2-42+ months) at the time of analysis. PMID- 12588357 TI - Long-term follow-up of granulocyte colony-stimulating factor receptor mutations in patients with severe congenital neutropenia: implications for leukaemogenesis and therapy. AB - Severe congenital neutropenia (SCN) is characterized by profound neutropenia, recurrent severe bacterial infections and maturation arrest in the myeloid lineage. Granulocyte colony-stimulating factor (G-CSF) treatment results in clinical improvement in over 90% of cases. Point mutations of the G-CSF receptor (G-CSFR) have been implicated in the progression of SCN to acute myeloid leukaemia (AML). Data are presented here on the 9-year follow-up of seven patients and the further screening of 18 other cases. One of the two original cases with a G-CSFR mutation has improved clinically; nevertheless, mutant DNA could still be detected at a very low level > 8 years after identification. The second child with a mutation progressed to myelodysplasia/AML 5 years after her mutation was detected. No mutations were found in the 18 new cases. One of three transformed cases had a G-CSFR mutation. This work is in agreement with the suggestion that G-CSFR mutations may provide a survival advantage to haemopoietic stem cells, but argues against the inevitability of leukaemic progression in their presence. Furthermore, the low frequency of G-CSFR mutations in SCN and the importance of regular screening and close clinical and laboratory follow-up if a mutation is found were demonstrated. PMID- 12588358 TI - Differential expression of CD2 on neoplastic mast cells in patients with systemic mast cell disease with and without an associated clonal haematological disorder. AB - Recently, aberrant coexpression of CD2 and CD25 has been reported to reliably distinguish neoplastic mast cells from normal or so-called reactive mast cells. Such expression is included in the consensus diagnostic criteria for systemic mast cell disease (SMCD). In our study of patients with SMCD, we found CD2 expression to be more prevalent on mast cells from patients without an associated haematological disorder (P = 0.04). Furthermore, no correlation was found between mast cell CD2 expression and other clinicopathological features in these patients. PMID- 12588359 TI - Angiogenesis occurs in hairy cell leukaemia (HCL) and in NOD/SCID mice transplanted with the HCL line Bonna-12. AB - Microvessel density (MVD), a surrogate marker for angiogenesis, was evaluated by anti-CD34 and CD105 monoclonal antibodies (Abs), and found to be increased in the bone marrow (BM) of hairy cell leukaemia (HCL) patients and in a preclinical model of non-obese diabetic/severe combined immunodeficient mice transplanted with the HCL line Bonna-12. The anti-CD105 Ab was significantly more sensitive than anti-CD34 Ab in identifying blood vessels. The BM tumour burden significantly decreased in patients treated with interferon-alpha, but the mean value of MVD remained unchanged. These data suggest that angiogenesis may be involved in the pathogenesis of HCL. PMID- 12588360 TI - Neutropenia and anaemia associated with T-cell large granular lymphocyte leukaemia responds to fludarabine with minimal toxicity. AB - T-cell large granular lymphocyte leukaemia (T-LGL) is a clonal disorder of T cells associated with neutropenia and anaemia. The clinical consequences are recurrent infections and transfusion dependence. The optimum treatment for severely affected patients remains to be defined. Current therapies require long term administration to maintain an effect. We report the reversal of severe neutropenia and/or anaemia in four patients treated with fludarabine which has been maintained since stopping treatment. The therapeutic side-effects were restricted to one episode of fever not associated with neutropenia. We conclude that fludarabine is effective in T-LGL, may be given safely despite severe neutropenia and induces durable treatment-free remissions. PMID- 12588361 TI - Changes of isoagglutinin titres after ABO-incompatible allogeneic stem cell transplantation. AB - We investigated the changes in isoagglutinin titres in 62 patients who underwent ABO-incompatible allogeneic stem cell transplantation. After major [and/or (+/-) minor] ABO-incompatible transplantation, recipient-derived isoagglutinins against donor-type red blood cells (RBCs) disappeared more rapidly in unrelated recipients (P = 0.006) and in patients with acute graft-versus-host disease (GVHD, P = 0.025) than in sibling recipients and in patients without acute GVHD respectively. Pure red cell aplasia (PRCA) developed in 10 out of 35 evaluable patients who underwent major (+/- minor) ABO-incompatible transplantation, and the post-transplant increase of isoagglutinin titres was a significant predictor for the occurrence of PRCA. In five out of 36 patients who underwent minor (and/or (+/-) major) ABO-incompatible transplantation, donor-derived isoagglutinins against recipient RBCs were detectable without clinically overt haemolysis. Isoagglutinin titres against ABO antigens absent both on recipient and donor RBCs decreased during the early post-transplant period then rose subsequently in 24 out of 29 patients at (median) d 59 post transplant. Our study showed that changes in isoagglutinin titres might have clinical implications in the occurrence of immunohaematological complications such as PRCA or immune mediated haemolysis, and might reflect immunohaematological reconstitution after transplantation. Furthermore, our data regarding time to disappearance of recipient-derived isoagglutinins against donor-type RBCs after major ABO incompatible transplantation suggest the presence of a graft-versus-plasma cell effect. PMID- 12588362 TI - Residual leukaemia after bone marrow transplant in children with acute lymphoblastic leukaemia after first haematological relapse or with poor initial presenting features. AB - Relapse is the major obstacle to cure for children with acute lymphoblastic leukaemia (ALL) after allogeneic bone marrow transplant (BMT). Development of salvage therapy for post-transplant relapse could be expedited by understanding the post-transplant behaviour of microscopically undetectable leukaemia and the ability to predict impending relapse. We have used a quantitative polymerase chain reaction method (sensitivity of 5.0 x 10-6) to measure residual leukaemia before the conditioning regimen, and at five time-points after transplantation. In total, 18 patients with ALL transplanted in first or second remission were studied for 1 year: For the first year post BMT, 12 remained in remission, four had haematological relapses, one had a cutaneous relapse, and one died of severe graft-versus-host disease. The post-engraftment levels of the leukaemia-specific immunoglobulin heavy (IgH) chain gene rearrangement for patients with haematological relapses were significantly different from those who remained in remission. The levels for the patients who remained in remission decreased with time, although there were occasional increases consistent with the known standard deviation of the measurement assay. In contrast, all clinical relapses were preceded by a rapid increase in levels. Both the rate of this increase and its timing were variable. These results suggest that residual leukaemia measurements can be used to direct post-transplant interventions and measure their effects. PMID- 12588363 TI - Heterozygotes in the bernard-soulier syndrome do not necessarily have giant platelets or thrombocytopenia. PMID- 12588367 TI - Quality of life measurements. PMID- 12588364 TI - What is the role of cytogenetic and molecular genetic analysis in the diagnosis of chronic myeloproliferative disorders? PMID- 12588368 TI - Basal cell carcinoma: a dermatopathological and molecular biological update. AB - The ideal classification of basal cell carcinoma (BCC) should be able to identify subtypes which correlate with clinical behaviour and treatment requirements. Unfortunately, however, such a classification has yet to be defined. In the interim, the currently most favoured classification is one based predominantly on histological growth pattern. This classification contributes to the useful concept of low- and high-risk histological subtypes of BCC. The latter are characterized by an increased probability of subclinical extension and/or incomplete excision and/or aggressive local invasive behaviour and/or local recurrence. The Royal College of Pathologists has published a minimum dataset for the histopathological reporting of BCC and this has been written to be compatible with the British Association of Dermatologists' management guidelines. Growth patterns to be reported include nodular, superficial, infiltrative/morphoeic and micronodular types, together with differentiation when of severely atypical or malignant squamous type (basosquamous carcinoma). Deep and peripheral excision margins will be reported to be either involved or clear. The latter will include a comment of a clearance of less than 1 mm for close margins and a measured distance in whole millimetres for other excisions. Clinical assessment and histology remain the 'gold standard' for evaluating BCC and cancers in general. However, in the postgenomic era emphasis is changing from the gathering and archiving of genomic data to its analysis and use in guiding clinical practice. In this context, a current goal is to define cancer phenotype in terms of molecular abnormalities and use this as a new gold standard. One way to assess whether this goal is being achieved for BCC is to determine whether our knowledge of its molecular pathology has any relevance to the minimum dataset for histological reporting. Knowledge of BCC molecular pathology has been fuelled by the recent discovery that deregulation of the Hedgehog (Hh) signalling pathway, a key player in embryonic patterning, appears to be fundamental to tumour growth. But despite accrual of a large amount of data concerning Hh pathway molecular alterations in neoplasia, little is known about the functional consequences of these changes in BCC, how they lead to tumour development, or how they relate to non-Hh pathway alterations such as TP53 mutation. Recent work suggests that the cellular localization of beta-catenin gives a degree of credence to the growth pattern classification of BCC. Furthermore, it is possible that beta-catenin may have a pathogenetic role in the invasive behaviour of BCC. This review draws on current evidence to discuss these issues and assess whether they are relevant to the minimum dataset. PMID- 12588369 TI - High level of thymus and activation-regulated chemokine in blister fluid and sera of patients with bullous pemphigoid. AB - BACKGROUND: Bullous pemphigoid (BP) is an autoimmune blistering disease characterized by eosinophilia and high serum IgE levels. The accumulated evidence suggests that various cytokines are involved in the lesional skin of patients with BP. Recently, thymus and activation-regulated chemokine (TARC/CCL17), a CC chemokine, was identified as a selective chemoattractant for CC chemokine receptor 4 (CCR4)-expressing cells. OBJECTIVE: In this study, we examined the involvement of TARC in patients with BP. METHODS: We determined the fluid and serum TARC levels in patients with BP by enzyme-linked immunosorbent assay and compared the serum TARC levels with the eosinophil numbers in peripheral blood. We also compared the serum TARC levels in five patients with BP before and after they were treated. Moreover, we examined TARC, CCR4 and CXC chemokine receptor 3 (CXCR3) expression in the lesional skin of patients with BP by immunohistochemical procedures. Furthermore, we measured CCR4 positivity in CD4+ CD45RO+ cells of peripheral blood mononuclear cells (PBMCs) in patients with BP and healthy control subjects. RESULTS: The fluid TARC levels in patients with BP were significantly higher than those in blisters from burn patients or suction blisters of healthy control subjects. The serum TARC levels in patients with BP were also significantly higher than those in pemphigus vulgaris (PV) patients and healthy control subjects, and decreased after the treatment. The serum TARC levels in patients with BP significantly correlated with the eosinophil numbers in peripheral blood (r = 0.72, P < 0.002). Immunohistochemistry showed a strong reactivity of TARC in the epidermal keratinocytes (KCs) of BP. Moreover, both CCR4 and CXCR3 were expressed on the dermal infiltrating CD4+ T cells mainly beneath the bullae of patients with BP. Fluorescence-activated cell sorting analysis showed a higher percentage of CCR4 positivity in CD4+ CD45RO+ cells of PBMCs in patients with BP than that in healthy control subjects, while there was no significant difference of CXCR3 positivity in CD4+ CD45RO+ cells of PBMCs between patients with BP and healthy control subjects. CONCLUSIONS: These findings strongly suggest that TARC may be one of the important chemokines that are involved in the pathogenesis of BP. PMID- 12588371 TI - Pattern analysis of human cutaneous mast cell populations by total body surface mapping. AB - BACKGROUND: Mast cells (MCs), critical effector cells in allergic inflammation and innate immunity to bacteria, are located in large numbers in tissues that interface the external environment, including the skin. However, little is known about the distribution and numbers of human skin MCs. OBJECTIVES: To assess the influence of age, sex and skin region on size and spatial distribution of MC populations in normal human skin. METHODS: Biopsies of healthy skin were obtained from 150 male and female individuals (age range 10-86 years). MCs were quantified and mapped planimetrically by histomorphometry in 15 anatomical sites (abdomen, thorax, lower and upper back, lower and upper arm, lower and upper leg, foot, hand, chin, nose, cheek, forehead, back of head). RESULTS: No differences in skin MC numbers or distribution were found when comparing skin obtained from male or female and from young or old individuals. At all skin sites, regardless of age or sex, MC numbers were highest in the most superficial skin layers where up to 10 fold more MCs were found as compared with the subcutis, which consistently contained the lowest numbers of MCs. Interestingly, MC numbers were highest at peripheral skin sites (maximum: chin and nose) and lowest at central skin sites (minimum: abdomen). Thus, healthy human skin exhibits a proximal/distal and a central/peripheral MC gradient and 'skin MC numbers' may vary by a factor of more than 20, depending on the skin layer and skin site analysed. CONCLUSIONS: These findings support the recently identified role of MCs in the elicitation of protective immune responses against infectious microorganisms. One possible explanation for these unexpected findings is that skin site/layer-specific factors (e.g. the density of nerves or vessels, exposure to ultraviolet irradiation, frequency of minimal trauma) are involved in the regulation of skin MC numbers and distribution. PMID- 12588370 TI - Selective cyclooxygenase-2 inhibition does not affect the healing of cutaneous full-thickness incisional wounds in SKH-1 mice. AB - BACKGROUND: The inducible cyclooxygenase-2 (COX-2) enzyme is upregulated in inflammatory diseases, as well as in epithelial cancers, and has an established role in angiogenesis and tissue repair. OBJECTIVE: Because of these physiological effects and the widespread use of the selective COX-2 inhibitor, celecoxib, we wanted to determine if inhibition of COX-2 would affect incisional skin wound healing. METHODS: Using a cutaneous full-thickness, sutured, incisional wound model in hairless SKH-1 mice, we evaluated the role of COX-2 in the wound healing process by comparing the effects of a nonselective COX inhibitor, diclofenac, with a selective COX-2 inhibitor, SC-791. Healing was monitored for up to 28 days postincision histologically and for recovery of wound strength. RESULTS: COX-2 expression was observed over the first week of healing, peaking at day 3 and was not affected by treatment with the selective COX-2 or nonselective COX inhibitors. Infiltrating macrophages, as well as keratinocytes and dermal fibroblasts at the wound site, expressed COX-2. Neither selective COX-2, nor nonselective COX inhibition had a significant effect on the macroscopic or microscopic morphology of the wounds, whereas dexamethasone treatment resulted in epidermal and granulation tissue atrophy. In addition, neither selective COX-2, nor nonselective COX inhibition altered keratinocyte proliferation and differentiation, dermal angiogenesis or the recovery of wound tensile strength, whereas dexamethasone reduced the tensile strength of the wounds by 30-38% throughout the healing period. CONCLUSIONS: These data indicate that selective COX-2 inhibition does not affect the healing of surgical skin wounds. PMID- 12588372 TI - Antimicrobial photodynamic therapy: assessment of genotoxic effects on keratinocytes in vitro. AB - BACKGROUND: Work has shown that cutaneous microbial species associated with skin conditions of microbial aetiology are susceptible to killing by antimicrobial photodynamic therapy (APDT) using visible light and methylene blue. OBJECTIVES: To evaluate immediate and delayed genotoxicity of APDT on keratinocytes in vitro. METHODS: A combination of methylene blue (100 microg mL(-1)) and visible light (42 mW cm(-2)), as used in studies of microbe and keratinocyte cytotoxicity, was employed to test a human keratinocyte cell line (H103) for genotoxic damage by comet assay. RESULTS: The comet assay was able to detect genotoxic damage in H2O2 treated keratinocytes (positive control). APDT did not cause either immediate or delayed genotoxic damage in keratinocytes following APDT of up to 180 min. CONCLUSIONS: APDT sufficient to reduce microbes by seven log cycles showed no detectable genotoxic effects on keratinocytes. APDT applied in vivo may represent a useful low-risk alternative to conventional antimicrobial treatment in dermatology. PMID- 12588373 TI - Psoriasis patients who are homozygous for the HLA-Cw*0602 allele have a 2.5-fold increased risk of developing psoriasis compared with Cw6 heterozygotes. AB - BACKGROUND: Psoriasis is strongly associated with certain human leucocyte associated antigens, especially HLA-Cw*0602. Patients who are HLA-Cw*0602 positive have been reported to have more active disease and a younger age at disease onset than HLA-Cw6-negative patients. OBJECTIVES: To ascertain whether there are differences in the clinical features and relative risk between HLA Cw*0602 homozygous and heterozygous psoriasis patients. METHODS: One thousand and six patients with chronic plaque psoriasis were evaluated clinically and HLA-C typed. In addition, 512 unrelated controls were typed for HLA-C. RESULTS: Of the patients 646 (64.2%) were HLA-Cw*0602 positive, and 68 (6.8%) were homozygous for this allele. Heterozygosity was associated with a relative risk of developing psoriasis of 8.9 compared with 23.1 for the Cw6 homozygous patients. The homozygous patients also had an earlier disease onset (mean 15.0 vs. 17.8 years, P = 0.04). However, the Cw6 homozygotes did not differ from the heterozygotes with respect to disease severity, guttate onset, distribution of plaques, nail changes or any other clinical parameter recorded. CONCLUSIONS: Homozygosity for the gene in the major histocompatibility complex region has a major additive impact on the risk of developing psoriasis and predisposes to an earlier disease onset, but does not have any marked influence on the phenotype or the severity of the disease. PMID- 12588374 TI - Wound healing effects of porcine placental extracts on rats with thermal injury. AB - BACKGROUND: Placental extracts have been used as Chinese folk medicines to accelerate wound healing. However, the molecular mechanism of placental extracts on wound healing has not been identified. It is known that fibroblast growth factors (FGF) and transforming growth factors (TGF) are two key factors involved in wound healing. OBJECTIVES: To determine the molecular mechanism of placental extracts on wound healing. METHODS: The protein levels of both growth factors in rat skins with thermal injury were therefore studied to explore the molecular mechanism of placental extracts on wound healing. As cell proliferation is essential for wound healing, effects of placental extracts on fibroblast proliferation were also determined. RESULTS: As compared with the controls, the S phase of fibroblasts was significantly increased by 1.5-, 1.7- and 4.7-fold for 1, 10 and 30 mg mL(-1) of placental extracts, respectively. The increase of the S phase was not due to the minute amount of sex hormones in the placental extracts as the addition of equivalent amounts of hormones showed no increase of the S phase. In addition, a 2.5-fold increase of TGF-beta1 in wound skin biopsy was noticed with 30 mg mL(-1) of porcine placental extracts. The FGF levels in the wound skin receiving 30 mg mL(-1) of porcine placental extracts were also significantly increased compared with the controls. CONCLUSIONS: These ex vivo data support the observation that the application of 30 mg mL(-1) of placental extracts reduced the wound healing time by about 50%. To the best of our knowledge, this is the first report to explore the molecular mechanisms of porcine placental extracts on wound healing. These results may provide the insight into the potential use of porcine placental extracts as an alternative medicine for accelerating wound healing. PMID- 12588375 TI - Rare occurrence of CD30+ circulating cells in patients with cutaneous CD30+ anaplastic large cell lymphoma: a study of nine patients. AB - BACKGROUND: The presence of a significant percentage of circulating atypical lymphocytes in peripheral blood has already been demonstrated in systemic CD30+ anaplastic large cell lymphoma (ALCL), which implies that a leukaemic component may be present in this subset of lymphomas. However, no similar data are available for the cutaneous counterpart of this particular lymphoproliferation. OBJECTIVES: To assess the presence of atypical cells, CD30+ lymphocytes and of a dominant T-cell clone in peripheral blood in a series of patients with cutaneous CD30+ ALCL. MATERIALS AND METHODS: Nine patients with either primary (four) or secondary (five) cutaneous CD4+ CD30+ ALCL were selected. The percentage of CD30+ CD4+ lymphocytes among peripheral blood mononuclear cells (PBMC) was determined by flow cytometry and the presence of a dominant circulating T-cell clone was assessed by polymerase chain reaction targeting the T-cell receptor gamma chain. A control group composed of apparently healthy individuals was similarly studied at the same time. RESULTS: The mean percentage of CD30+ cells in PBMC was slightly higher in patients than in controls (3.9% vs. 2.7%) but the difference was not statistically significant. Only two patients displayed more than 5% CD30+ cells, both of whom had a minor tumour burden. A dominant circulating T-cell clone was detected in only three cases, including these two latter patients. CONCLUSIONS: The occurrence of a significant percentage of CD30+ CD4+ circulating cells is rare in active cutaneous CD30+ ALCL, either primary or secondary. This percentage is not related to the apparent skin tumour burden but a significant figure appeared to be correlated with the detection of a dominant T-cell clone in peripheral blood. Overall, these data show that, unlike mycosis fungoides, peripheral blood involvement seems infrequent in cutaneous CD30+ ALCL. The hypothesis that a high percentage of CD30+ circulating cells might be related to the presence of a cryptic systemic disease cannot be ruled out. PMID- 12588376 TI - Comparison of diagnostic accuracy for cutaneous malignant melanoma between general dermatology, plastic surgery and pigmented lesion clinics. AB - BACKGROUND: Since the 1980s there have been dedicated pigmented lesion clinics (PLCs) in the U.K. Important considerations when comparing the efficacy of the PLC with other referral clinics include diagnostic accuracy. OBJECTIVES: To compare the false-negative rate of clinical diagnosis (FNR) in the PLC with that in the other clinics of primary referral of malignant melanoma (MM) in the same geographical area. We have previously shown that certain clinical features are risk factors for diagnostic failure of MM. A further aim of this study was to correct for any differences in frequency of these factors in the melanoma populations between clinics and to estimate the false-positive diagnostic rate (FPR) in the PLC. METHODS: To compare the FNR between clinics, the case notes of all patients presenting with histologically proven cutaneous MM in Leicestershire between 1987 and 1997 were examined retrospectively. A false-negative diagnosis was defined as documentation of another diagnosis and/or evidence in the case notes that the diagnosis was not considered to be MM. The FNR was estimated as the number of false-negative clinical diagnoses/number of true-positive histological diagnoses. To estimate the diagnostic FPR, which was defined as the number of false-positive clinical diagnoses of MM/total number of positive clinical diagnoses, in the PLC, the outcome of 500 consecutive patients attending the PLC was surveyed. RESULTS: The case notes of 731 patients were available, of whom approximately two-thirds initially attended the PLC, one-fifth the General Dermatology clinics (D) and the remainder were divided approximately equally (one twentieth each) between Plastic Surgery clinics (P), other clinics (O) and the surgery of the general practitioner (GP). The last was regarded as the primary referral clinic if the lesion were excised there prior to any referral. The FNR was lowest for the PLC, at 10%, compared with 29% (D), 19% (P), 55% (O) and 54% (GP) (P < 0.0001). Lesions with risk factors for diagnostic failure were under represented in the PLC (P < 0.0001), the mean frequencies of the risk factors being 20% (PLC), 25% (D), 22% (P), 31% (O) and 30% (GP). Differences were not large but still could partially explain the lower FNR of the PLC. However, when the FNR was estimated for lesions exhibiting each of these risk factors, the PLC was found to have the lowest rate in every case (PLC vs. all clinics combined, P = 0.04 to P < 0.0001). The mean FNR for the risk factors combined was 18% (PLC), 45% (D), 50% (P), 68% (O) and 71% (GP). Also on logistic multivariable analysis of the PLC vs. all the other clinics on FNR and the above factors, the higher FNR of the other clinics retained significance (odds ratio 5.9, P < 0.0001). In the 500 patients surveyed separately in the PLC, the MM pick-up rate on biopsy was 32% and the diagnostic FPR was 41%. CONCLUSIONS: The FNR of MM was lower in the PLC than in the other clinics, while the pick-up rate for MM on biopsy and the FPR were acceptably low. PMID- 12588377 TI - The British standard series of contact dermatitis allergens: validation in clinical practice and value for clinical governance. AB - BACKGROUND: All centres use an empirically determined set of 'standard' test allergens for patch testing that contain the commoner environmental sensitizers. Objectives To assess the validity of the British standard series of 12 allergens used in addition to the 23 already in the European standard series. PATIENTS AND METHODS: Results for 3062 consecutive patients patch tested in seven centres across the United Kingdom during the year 2000 were analysed. RESULTS: The additional allergens from the British series and positive rates were: methyl dibromoglutaronitrile 2.4%, carba mix 1.6%, tixocortol pivalate 1.5%, ethylenediamine 1.3%, cetearyl alcohol 0.8%, 2-bromo-2-nitropane-1,3-diol 0.8%, diazolidinyl urea 0.7%, chlorocresol 0.6%, budesonide 0.6%, fusidic acid 0.5%, imidazolidinyl urea 0.5%, and chloroxylenol 0.4%. The allergens with the lowest positive rate in the European standard series were primin at 0.6% and isopropyl phenyl-para-phenylenediamine at 0.4%. CONCLUSIONS: The 12 allergens in the British series should continue being tested as a standard addition to the European series within the U.K. The collection of data in this manner to allow comparisons between centres shows differences that reflect selection criteria and interpretation of results, and offers a useful tool for audit and clinical governance. Testing fewer than 1 : 2150 population may indicate underprovision of service. Similarly, rates of sensitization for nickel contact allergy above 26% and for fragrance mix above 16% (the upper 95% confidence intervals) should stimulate inquiry into the reasons behind this. PMID- 12588378 TI - Minimal residual disease in mycosis fungoides follow-up can be assessed by polymerase chain reaction. AB - BACKGROUND: T-cell receptor (TCR) gene rearrangement analysis, i.e. T-cell clonality, using polymerase chain reaction (PCR) is a routine method used to assess the presence of a cutaneous dominant T-cell clone in mycosis fungoides (MF). OBJECTIVES: To compare the outcome of cutaneous lesions of MF after treatment with the fate of the cutaneous T-cell clonality, and to determine whether minimal residual disease can be detected in patients in clinical complete remission. METHODS: Fifty-one patients histologically diagnosed as having MF (17 stage IA, 21 stage IB and 13 stage III) were included in this retrospective study. T-cell clonality was analysed by GC-clamp multiplex PCRgamma-denaturing gradient gel electrophoresis. Every patient had two cutaneous biopsies at least 3 months apart. The second biopsy was performed at the site of a treated lesion. RESULTS: The presence or absence of a dominant T-cell clone in the skin remained identical in 26 of the 31 (84%) patients with persistent disease. Thirteen patients with a detectable dominant T-cell clone at diagnosis went into complete clinical remission. In nine of these 13 (69%) patients, the T-cell clone was no longer detectable after treatment. The remaining four (31%) patients had an unchanged T-cell clonality. CONCLUSIONS: The TCR gene rearrangement imprint is a stable and reliable tumour marker of MF disease. One-third of patients in complete clinical remission had a cutaneous molecular residual disease, the prognostic value of which will be analysed in an ongoing prospective study. PMID- 12588379 TI - Computerized morphometry and three-dimensional image reconstruction in the evaluation of scalp biopsy from patients with non-cicatricial alopecias. AB - BACKGROUND: A major challenge in the histopathological examination of scalp biopsies is to perform an adequate evaluation of all the hair follicles present in the tissue. Transverse sectioning is currently the preferred technique to demonstrate every follicular structure in a punch biopsy specimen, although diagnostic accuracy is dependent on subjective evaluation of follicular morphology and hair size. OBJECTIVES: To determine if computer-based morphometry and three-dimensional (3D) image reconstruction software can be used to evaluate scalp biopsies from patients with non-cicatricial alopecias. METHODS: Nine 4-mm scalp punches were taken from nine patients with noncicatricial alopecias and step-sectioned transversely at 0.1-mm intervals from the epidermal surface to the subcutaneous fat. Each tissue section was then digitized and analysed using morphometric and 3D image reconstruction software. Morphometric data and 3D images were collated with clinical and conventional light microscopic diagnoses, as well as follow-up information. RESULTS: In four of the nine patients, results of morphometric analysis concurred with conventional clinicopathological diagnoses. In the remaining five patients, morphometry revealed a lower telogen count in one patient and higher telogen count in four patients. One of the four patients with a higher telogen count also had a low mean hair diameter and miniaturized anagen follicles in the 3D image that were suggestive of early androgenetic alopecia (AGA). 3D virtual microscopic imagery allowed the direct visualization of colour-coded, scaled hair follicles which demonstrated characteristic changes in alopecia areata, AGA and telogen effluvium. CONCLUSIONS: Our study demonstrated the feasibility of using morphometric and 3D reconstruction software to evaluate scalp biopsies. With further validation, this technique may prove to be more sensitive to detect subtle quantitative and qualitative follicular changes in non-cicatricial alopecias. PMID- 12588380 TI - Dermatology Life Quality Index: influence of an illustrated version. AB - BACKGROUND: An illustrated version of the Dermatology Life Quality Index (DLQI) was developed in order to improve the ease with which this standard questionnaire is answered. OBJECTIVES: To compare the illustrated version with the text-only version of the DLQI. METHODS: The two versions were administered to 206 patients attending Dermatology outpatient clinics. The time taken to complete either the illustrated or the text-only version was noted in 52 additional patients. RESULTS: One hundred and ninety-one of 206 patients completed both questionnaires satisfactorily. Their mean DLQI scores were 6.4 (median 4.0, lower quartile 1.0, upper quartile 11.0) for the illustrated version and 6.3 (median 4.0, lower quartile 1.0, upper quartile 10.0) for the text-only version. For the 98 patients who answered the illustrated version first, the mean DLQI scores were 6.9 (median 4.0, lower quartile 1.9, upper quartile 11.9) for the illustrated version and 6.4 (median 4.0, lower quartile 1.8, upper quartile 11.0) for the text-only version. For the 93 patients who answered the text-only version first, the mean DLQI scores were 5.9 (median 4.0, lower quartile 1.7, upper quartile 10.2) for the illustrated version and 6.2 (median 5.0, lower quartile 0.5, upper quartile 9.5) for the text-only version. The median time taken to complete the text-only version was 124 s (mean +/- SD 126 +/- 65, n = 27) and 88 s (mean +/- SD 101 +/- 52, n = 25) for the illustrated version (P = 0.08). There was a very close correlation (r = 0.98) between the scores of the text-only and the illustrated versions; 82.2% of patients either scored the same or had a DLQI score difference of only 1. One hundred and seven (59.8%) patients preferred using the illustrated version and 66 (36.9%) preferred the text-only version. One hundred and four (57.5%) found the illustrated version easier to use. CONCLUSIONS: The illustrated version was superior to the text-only version in terms of patient preference and ease of use, but it was not possible to demonstrate exact equivalence between the two versions. PMID- 12588381 TI - The Children's Dermatology Life Quality Index: validation of the cartoon version. AB - BACKGROUND: In 1995 the Children's Dermatology Life Quality Index (CDLQI) was developed as a tool to allow quality of life assessment of children with skin conditions. This initial questionnaire was in a written format. OBJECTIVES: Using the same validated questions, a full-colour cartoon version has been developed. The aim of this study was to validate this against the initial written questionnaire in a three-part study. METHODS: The first part of the study piloted the use of both versions in an outpatient setting. One hundred and one children completed both versions of the CDLQI in a random order. A further 66 children completed the cartoon CDLQI in outpatients, and subsequently completed the cartoon version on the same day at home, which was returned by post. The scores were compared. In the second part, in more controlled conditions to eliminate parental and investigator bias, 107 children with current dermatological problems were administered both versions of the CDLQI in a random order. The scores were analysed, and time to complete each version, and the child and parental preferences, were recorded. The third part assessed compliance by asking 546 children recently reviewed in dermatology clinics to return a single completed postal CDLQI. Half of the children were given the text, and half the cartoon version. RESULTS: The median age of participating children was 11 years. There was no significant difference in scores between the two versions in both parts 1 and 2, but the cartoon version was completed faster (median 90 s) than the written version (median 120 s) (P < 0.0001). Both children and their parents significantly preferred the cartoon version and found it easier to use. Forty-six per cent of the postal CDLQI questionnaires were returned; there was no difference in compliance between the two versions. CONCLUSIONS: The cartoon CDLQI is equivalent to the previously validated written CDLQI version, but is faster and easier for children to use, and is preferred by both children and parents. PMID- 12588382 TI - Alterations in human epidermal Langerhans cells by ultraviolet radiation: quantitative and morphological study. AB - BACKGROUND: Ultraviolet (UV) exposure of human skin induces local and systemic immune suppression. This phenomenon has been well documented when UVB radiation (290-320 nm) is used. The mechanism is thought to involve Langerhans cells (LCs), the epidermal dendritic cells that play a crucial role in antigen presentation. A variety of studies have clearly demonstrated that UVB radiation decreases LC density and alters their morphology and immunological function, but little is known about the effects of the entire UV spectrum (ultraviolet solar simulated radiation, UV-SSR or UVB + UVA) or UVA (320-400 nm) radiation alone. OBJECTIVES: The purpose of this study was to analyse and compare the effects of a single exposure of human volunteers to UV-SSR, total UVA or UVA1 (340-400 nm) in the human epidermal LC density and morphology. METHODS: Immunohistochemistry on epidermal sheets with various antibodies and transmission electron microscopy (TEM) were used. RESULTS: Immunostaining for class II antigen revealed that a single UV-SSR exposure, corresponding to twice the minimal erythemal dose (MED), induced a significant reduction in LC density with only slight morphological alterations of remaining cells. After a single UVA exposure, LC density showed a dose-dependent reduction with a significant effect at 60 J cm(-2) (well above the MED). Moreover, the reduction of LC dendricity was also dose-dependent and significant for doses exceeding 30 J cm(-2). UVA1 radiation was as effective as total UVA for the later endpoint. As demonstrated by TEM, the location of Birbeck granules containing epidermal cells was modified in UVA-exposed areas. They were located in the spinous rather than in the suprabasal layer. In addition, the morphology of these cells was altered. We observed a rounding up of the cell body with a reduction of dendricity. Alterations of mitochondrial membrane and ridges were also seen. CONCLUSIONS: A single exposure of human skin in vivo to UV-SSR, UVA or UVA1 radiation results in different alterations of density and/or morphology of LCs. All these alterations may impair the antigen-presenting function of LCs leading to an alteration of immune response. PMID- 12588383 TI - The effect of ultraviolet radiation exposure on the prevalence of mast cells in human skin. AB - BACKGROUND: Dermal mast cells have been implicated as important effector cells in innate immunity, hypersensitivity responses and ultraviolet (UV)B-induced suppression of cell-mediated immune responses to contact allergens. Humans, like mouse strains, display variations in dermal mast cell prevalence. The factors determining these differences are yet to be fully elucidated. In mice, expression of the receptor for stem cell factor, c-kit, on dermal mast cells correlates with prevalence. OBJECTIVES: To evaluate dermal mast cell prevalence and mast cell c kit expression in non-sun-exposed and sun-exposed skin in the same donor. METHODS: In 14 subjects, biopsies of skin (4 mm) were sampled from the skin sites of buttock, inner arm, shoulder and back of hand skin and dermal mast cell prevalence quantified. Non-sun-exposed buttock and chronically sun-exposed hand skin were evaluated for mast cell expression of c-kit and elastin content, a feature of photoageing and surrogate marker of UV exposure. RESULTS: The prevalence of dermal mast cells was significantly higher in hand skin than in the three other anatomically different skin sites. Significant correlations were observed in hand but not buttock skin between increasing dermal mast cell densities, extent of elastin content in the papillary dermis and age of the subject. Cellular expression of c-kit correlated with mast cell prevalence in hand skin. However, no relationship was observed in hand skin between c-kit expression, elastin content and age. CONCLUSIONS: The prevalence of mast cells in human skin is altered by factors that are intrinsic (mechanisms regulating c-kit expression) and extrinsic (chronic sun exposure), and the fact that the associations of mast cell prevalence with age is explained by the latter being a correlate of cumulative sun exposure. PMID- 12588384 TI - Randomized, double-blind, placebo-controlled trial of autologous blood therapy for atopic dermatitis. AB - BACKGROUND: Autologous blood therapy (ABT) is used for treating atopic dermatitis (AD) in some European countries and is promoted on internet sites for this condition. However, there is little evidence from rigorous clinical trials to suggest that it is effective. OBJECTIVES: To test the effectiveness of ABT for the symptomatic treatment of patients with AD. METHODS: Fifty subjects responded to press advertisements, and 31 were randomized within strata of severity at recruitment. Patients were included into a double-blind, placebo-controlled trial and received ABT or placebo once weekly for 5 weeks. Assessments were performed at baseline, at weekly intervals and after a 5-week follow up. The Six Area, Six Sign AD (SASSAD) severity index was predefined as the primary outcome measure. The Dermatology Life Quality Index and patient ratings of pruritus, quality of sleep and skin appearance on 100-mm visual analogue scales were defined as secondary outcome measures. Success of patient blinding and adverse events were assessed. RESULTS: Data were analysed on an intention-to-treat basis. Analysis of covariance suggested a significant differential change of the SASSAD score between baseline and the end of the follow-up period in favour of ABT. The mean reduction in SASSAD score was 13.5 points (95% confidence interval, CI 6.6-20.4, P < 0.001) over and above placebo; the corresponding value at the end of treatment was 9.6 (95% CI 4.2-14.9, P = 0.001). No clear significant intergroup differences in any of the secondary outcome measures were found. Six patients in the ABT group and seven in the placebo group reported minor and transient adverse events. CONCLUSIONS: These data suggest that, according to the SASSAD score, ABT has beneficial effects in the treatment of AD, although this was not confirmed by the patient-rated assessments. The improvement in observer-rated skin condition suggested by this study needs confirmation in larger trials. PMID- 12588385 TI - A randomized, placebo-controlled, double-blind trial of ondansetron in renal itch. AB - BACKGROUND: Renal itch is a relatively common and distressing problem for patients with chronic renal failure. Ondansetron, a serotonin type 3 receptor antagonist was developed for relief of chemotherapy induced nausea. Recently, anecdotal reports describe relief of renal itch with ondansetron. OBJECTIVES: We performed a double-blind randomized placebo-controlled trial to objectively assess the effectiveness of ondansetron in renal itch. PATIENTS AND METHODS: With approval from the local ethical committee, 24 patients on haemodialysis were enrolled in the trial. On a random basis 14 patients were blindly allocated to the ondansetron-placebo sequence and 10 to the placebo-ondansetron sequence. Baseline values for itch were obtained for 7 days before the treatment period and there was a 7-day washout between the treatment periods. During the treatment patients received either 8 mg of ondansetron three times a day or a placebo tablet three times a day for 2 weeks. Patients were asked to record the severity of their pruritus on a visual analogue scale (VAS) twice a day. At the end of the study patients were asked blindly which treatment they had preferred. RESULTS: Seventeen patients completed the trial. Pruritus decreased by 16% (95% CI: 0.5 32%) during active treatment and by 25% (95% CI: 9-41%) during treatment with placebo. The change in VAS scores during treatment with ondansetron (P = 0.04) and placebo (P = 0.01) were both significant. Eleven patients expressed a preference, seven for placebo and four for ondansetron. CONCLUSIONS: Our results show that ondansetron is no better than placebo in controlling renal itch. PMID- 12588386 TI - The combination of calcipotriol and methotrexate compared with methotrexate and vehicle in psoriasis: results of a multicentre placebo-controlled randomized trial. AB - BACKGROUND: A multicentre, randomized, double-blind, vehicle-controlled, parallel group study was carried out to study the effect of the addition of calcipotriol ointment to methotrexate (MTX) therapy in patients with psoriasis vulgaris. OBJECTIVES: To investigate whether the addition of calcipotriol to treatment with MTX has an MTX-sparing effect, and whether the combination of treatments is safe. Additionally, to compare the effect of calcipotriol or vehicle on the duration of the relapse-free interval after cessation of MTX. METHODS: Patients on maintenance therapy with MTX with controlled psoriasis were selected. The study was divided into three phases: (i) an MTX-free phase with double-blind treatment with either calcipotriol ointment or vehicle; (ii) an MTX titration phase with open MTX treatment and additional double-blind treatment with either calcipotriol or vehicle until target response; and (iii) follow-up phase: in a group of 97 patients, psoriasis was assessed using the modified psoriasis severity score, patients' assessment and safety parameters were monitored as well. RESULTS: The combined use of calcipotriol with MTX resulted in an MTX-sparing effect of 3.4 mg week-1 (phase (II) and 2.6 mg week-1 (phase I and II taken together), while still maintaining efficacy. Calcipotriol treatment increased the time to relapse of psoriasis following discontinuation of MTX: 113 days vs. 35 days. A decrease in aspartate aminotransferase and alanine aminotransferase was seen during the study of 8% (calcipotriol) and 12% (vehicle). CONCLUSIONS: The combination of calcipotriol and MTX was safe and well tolerated. The combination resulted in lower cumulative dosages of MTX compared with MTX and vehicle. Therefore the risk of side-effects is substantially decreased. PMID- 12588387 TI - Intra-individual comparison of the cutaneous safety and efficacy of calcitriol 3 microg g(-1) ointment and calcipotriol 50 microg g(-1) ointment on chronic plaque psoriasis localized in facial, hairline, retroauricular or flexural areas. AB - BACKGROUND: Psoriasis involving sensitive skin areas remains difficult to treat because of the side-effects of topical corticosteroids and the irritancy potential of vitamin D3 derivatives. Several clinical trials have demonstrated that calcitriol, the naturally occurring and hormonally active form of vitamin D3, is effective and safe at the dose of 3 microg g(-1) for the treatment of psoriasis affecting the trunk and limbs. METHODS: We compared the safety and efficacy of calcitriol 3 microg g(-1) ointment and calcipotriol 50 microg g(-1) ointment in a multicentre, randomized, investigator-blinded, left-right comparison in mild to moderate chronic plaque psoriasis affecting sensitive areas, defined as being the face, hairline, retroauricular and flexural areas. One pair of symmetrical and bilateral target lesions was selected from each area and assessed for perilesional erythema, oedema, and stinging/burning. Global assessment of local tolerability and global improvement were rated by the investigator, and the subjects were asked to evaluate the tolerability and efficacy of each product and to express their global preference. RESULTS: In the 75 subjects, calcitriol and calcipotriol both led to clearing of at least one target lesion in 21 (28%) of the subjects each. Perilesional erythema (P < 0.001), perilesional oedema (P < 0.02) and stinging/burning (P < 0.001) were all significantly less severe with calcitriol than with calcipotriol. The subjects' evaluation of local tolerability was significantly (P < 0.0001) in favour of calcitriol. Ten treatment-related dermatological events occurred in eight subjects, including one subject who experienced skin discomfort on both sides. All other events occurred only on the calcipotriol-treated side (irritant dermatitis, six subjects; contact dermatitis, one subject). Global assessment of improvement from baseline by the investigators was significantly greater for the calcitriol-treated lesions (P < 0.02). The subjects' global preference was significantly in favour of calcitriol (P < 0.02). CONCLUSIONS: In the present study, calcitriol ointment was found to be better tolerated and would appear to be more effective than calcipotriol ointment in the treatment of psoriasis in sensitive areas. PMID- 12588388 TI - Direct stimulatory effect of low-intensity 670 nm laser irradiation on human endothelial cell proliferation. AB - BACKGROUND: Endothelial cell (EC) proliferation plays a key role in the process of tissue repair. Low-intensity laser irradiation has been demonstrated to accelerate wound healing and to improve microvascularization. OBJECTIVES: The present study evaluated a possible stimulatory influence of low-intensity laser irradiation on human umbilical vein endothelial cell (HUVEC) proliferation in a systematic manner. METHODS: Subconfluent cultures of HUVEC were irradiated every other day with a 670-nm diode laser (intensity: 10-65 mW cm(-2), dose: 2-8 J cm( 2)) during a period of 6 days. Cell proliferation was evaluated quantitatively by counting in a haemocytometer. RESULTS: Our data demonstrate a dose-dependent and intensity-dependent stimulatory effect of laser irradiation on HUVEC cell proliferation. Doses of between 2 and 8 J cm(-2) induced statistically significant cell proliferation. Testing different intensities at a constant dose of 8 J cm(-2), 20 and 65 mW cm(-2) induced most pronounced cell proliferation. CONCLUSIONS: Low-intensity laser irradiation influences EC proliferation and might thereby contribute to the increase in angiogenesis and the acceleration of wound healing in vivo. PMID- 12588389 TI - A possible paracrine hedgehog signalling pathway in neurofibromas from patients with neurofibromatosis type 1. AB - BACKGROUND: Hedgehogs (Hhs) and their receptors are involved in organ development as well as in tumorigenesis observed in basal cell carcinoma. Among Hhs, Desert hedgehog secreted from Schwann cells mediates the formation of peripheral nerve sheaths. However, there has been no study on the role of Hhs and their receptors in tumorigenesis of neurofibromas in neurofibromatosis type 1 (NF1). OBJECTIVES: To clarify the expression and localization of Hhs and their receptors in neurofibromas of NF1 patients. METHODS: Expression of Hhs and their receptors was studied by immunohistochemistry using neurofibromas from NF1 patients and control normal skin samples. RESULTS: In neurofibromas, CD57-positive tumour cells with delicate elongated processes were positive for the receptor PTCH2. Perineurial cells of involved nerves within neurofibromas as well as those of normal cutaneous nerves expressed Indian hedgehog and Sonic hedgehog. Schwann cells of normal cutaneous nerves were positive for PTCH2. CONCLUSIONS: Our study suggests that a paracrine Hh signalling pathway may be involved in tumorigenesis of neurofibromas in NF1. PMID- 12588390 TI - Cutis marmorata telangiectatica congenita and extensive mongolian spots: type 5 phacomatosis pigmentovascularis. AB - Two patients with an unusual association of extensive cutis marmorata telangiectatica congenita and aberrant mongolian spots are reported. We believe that such association is best explained as a phenomenon of non-allelic twin spotting. We found only one previous similar case in a textbook, and we support the classification of this condition as phacomatosis pigmentovascularis type V. PMID- 12588391 TI - Erythroderma: an unusual presentation of pulmonary tuberculosis. AB - We report an elderly Chinese man who presented with erythroderma. He was treated with topical corticosteroids and emollients, but did not improve. Chest X-ray revealed cavitation in the right upper pulmonary lobe, and laryngeal swab culture was positive for Mycobacterium tuberculosis complex. Polymerase chain reaction performed on a skin biopsy was positive for M. tuberculosis DNA. Within a month of starting antituberculous medication, his erythroderma had almost completely resolved. Pulmonary tuberculosis should be included in the list of differential diagnosis for erythroderma. PMID- 12588392 TI - Erythrodermic syringotropic cutaneous T-cell lymphoma. AB - Syringotropic cutaneous T-cell lymphoma (CTCL) is a rare localized variant of CTCL, characterized histologically by eccrine gland and ductal hyperplasia surrounded by a dense syringotropic lymphocytic infiltrate. Previously reported only in men, we describe the first woman with syringotropic CTCL. Unusually, she presented with erythroderma, cutaneous nodules, poikilodermatous patches, widespread alopecia and lymphadenopathy. PMID- 12588393 TI - Efficacy of topical tacrolimus for treating the malar rash of systemic lupus erythematosus. AB - The rash of systemic lupus erythematosus (SLE) is usually treated with topical corticosteroids, but prolonged use causes adverse cutaneous side-effects. We assessed the efficacy of topical tacrolimus for treating the skin lesions of SLE. Three patients with SLE affecting their facial skin applied 0.1% tacrolimus ointment on one side of their face twice daily for 3 weeks, in conjunction with a sunscreen cream. After 3 weeks, erythema on the treated side was ameliorated in all three patients compared with the untreated side. Although the study is preliminary, the results demonstrate that topical tacrolimus may be useful for treating the malar rash of SLE. PMID- 12588394 TI - Nodular sclerotic change in dermatofibrosarcoma protuberans: a potential diagnostic problem. AB - Nodular sclerotic change in dermatofibrosarcoma protuberans (DFSP) is not a well recognized event. Sclerotic change is reported in three patients with DFSP: a 33 year-old woman with a left subclavian mass, a 29-year-old woman with a mass on the left crus and a 31-year-old man with a mass on the left cheek. Histologically, lesions consisted of two types of feature, one type being typical of DFSP and the other being rich in collagen. In the first patient, each type formed a distinct compartment, each of which contained scattered cells containing melanin pigment. In the second patient, foci with typical DFSP histology were distributed within the collagen-rich area and also formed a substantial compartment in the periphery of the lesion, with pigmented cells being distributed mainly in and around the cellular area. In the third patient, the collagenous area occupied about half of the lesion. There was no pigmentation. In all three cases the collagen-rich area contained sparsely distributed fibroblast like cells that stained positive with CD34. Sclerotic change in DFSP could cause substantial difficulty in diagnosis if the biopsy material were partial and small. The clinical history and knowledge about the existence of this kind of histological variation in DFSP are important. PMID- 12588395 TI - An unusual clinical pattern of cutaneous metastasis: target-like lesions. PMID- 12588396 TI - Cheilitis glandularis: a rare entity. PMID- 12588397 TI - An unusual variant of granulomatous adnexotropic cutaneous T-cell lymphoma. PMID- 12588398 TI - Comparison of melanocytic naevi with photographs, a recommended method. PMID- 12588399 TI - Antibiotic-resistant Propionibacteria acnes may not be the major issue clinically or microbiologically in acne. PMID- 12588400 TI - Propionibacterial biofilms cannot explain antibiotic resistance but might contribute to some cases of antibiotic recalcitrant acne. PMID- 12588401 TI - Squamous cell carcinoma in a patient with non-bullous congenital ichthyosiform erythroderma. PMID- 12588402 TI - A case of subcutaneous sparganosis: therapeutic assessment by an indirect immunofluorescence antibody titration using sections of the worm body obtained from the patient. PMID- 12588403 TI - Audit of basal cell carcinoma: registration practice. PMID- 12588404 TI - Late (> 10 years) recurrence of melanoma: the Scottish experience. PMID- 12588405 TI - Dermatology Life Quality Index score in vitiligo and its impact on the treatment outcome. PMID- 12588406 TI - Oral lichen planus: diagnostic immunofluorescence testing on routine histological material. PMID- 12588407 TI - Severe pruritus in a haemodialysed patient: dramatic improvement with granisetron. PMID- 12588408 TI - Borrelia afzelii evidenced by polymerase chain reaction in a biopsy of nipple lymphocytoma. PMID- 12588409 TI - Arterial embolization caused by injection of hyaluronic acid (Restylane). PMID- 12588411 TI - Prevalence of hypertension among patients with atopic dermatitis. PMID- 12588413 TI - The early erosive vesicular stage of lipoid proteinosis: clinical and histopathological features. PMID- 12588416 TI - The role of bacteriolysis in the pathophysiology of inflammation, infection and post-infectious sequelae. AB - The literature dealing with the biochemical basis of bacteriolysis and its role in inflammation, infection and in post-infectious sequelae is reviewed and discussed. Bacteriolysis is an event that may occur when normal microbial multiplication is altered due to an uncontrolled activation of a series of autolytic cell-wall breaking enzymes (muramidases). While a low-level bacteriolysis sometimes occurs physiologically, due to "mistakes" in cell separation, a pronounced cell wall breakdown may occur following bacteriolysis induced either by beta-lactam antibiotics or by a large variety of bacteriolysis inducing cationic peptides. These include spermine, spermidine, bactericidal peptides defensins, bacterial permeability increasing peptides from neutrophils, cationic proteins from eosinophils, lysozyme, myeloperoxidase, lactoferrin, the highly cationic proteinases elastase and cathepsins, PLA2, and certain synthetic polyamino acids. The cationic agents probably function by deregulating lipoteichoic acid (LTA) in Gram-positive bacteria and phospholipids in Gram negative bacteria, the presumed regulators of the autolytic enzyme systems (muramidases). When bacteriolysis occurs in vivo, cell-wall- and -membrane associated lipopolysaccharide (LPS (endotoxin)), lipoteichoic acid (LTA) and peptidoglycan (PPG), are released. These highly phlogistic agents can act on macrophages, either individually or in synergy, to induce the generation and release of reactive oxygen and nitrogen species, cytotoxic cytokines, hydrolases, proteinases, and also to activate the coagulation and complement cascades. All these agents and processes are involved in the pathophysiology of septic shock and multiple organ failure resulting from severe microbial infections. Bacteriolysis induced in in vitro models, either by polycations or by beta lactams, could be effectively inhibited by sulfated polysaccharides, by D-amino acids as well as by certain anti-bacteriolytic antibiotics. However, within phagocytic cells in inflammatory sites, bacteriolysis tends to be strongly inhibited presumably due to the inactivation by oxidants and proteinases of the bacterial muramidases. This might results in a long persistence of non biodegradable cell-wall components causing granulomatous inflammation. However, persistence of microbial cell walls in vivo may also boost innate immunity against infections and against tumor-cell proliferation. Therapeutic strategies to cope with the deleterious effects of bacteriolysis in vivo include combinations of autolysin inhibitors with combinations of certain anti inflammatory agents. These might inhibit the synergistic tissue- and- organ damaging "cross talks" which lead to septic shock and to additional post infectious sequelae. PMID- 12588417 TI - Soluble membrane-type 1 matrix metalloproteinase (MT1-MMP) and gelatinase A (MMP 2) in induced sputum and bronchoalveolar lavage fluid of human bronchial asthma and bronchiectasis. AB - The aim of this study was to investigate the involvement of the MT1-MMP/MMP-2 cascade in induced sputum (IS) and bronchoalveolar lavage fluid (BALF) from bronchial asthma (BA) and bronchiectasis (BE) patients and healthy controls. The molecular forms and cellular origins of MT1-MMP and MMP-2 were determined by Western immunoblotting, immunohistochemistry and in situ hybridization. Elevated levels of soluble activated and autocatalyzed MT1-MMP species as well as activated forms of MMP-2 in IS and BALF samples from BA and BE patients were evidenced. The activation degrees of soluble MT1-MMP and MMP-2 were significantly correlated in BA and BE IS and BALF. Only low levels of both these MMPs were observed in healthy control IS and BALF. The co-expression of MMP-2 with MT1-MMP was evidenced by double immunostaining in bronchial epithelial cells, submucosal glandular cells, smooth muscle cells and monocyte/macrophages. The MT1-MMP/MMP-2 cascade is present and active in human inflammatory lung disease fluid and tissue samples. This cascade seemingly reflects the active destructive phases of these chronic lung diseases. PMID- 12588418 TI - Morules with optically clear nuclei in ovarian borderline endometrioid tumor. AB - Optically clear nuclei (OCN) have been observed in morules of some neoplasms and in some conditions unrelated to the development of the morules. We first report a case of ovarian borderline endometrioid tumor (BET) showing the morules associated with OCN. The patient was a 47-year-old premenopausal woman with a left ovarian cystic tumor, atypical endometrial hyperplasia, and elevated serum levels of FSH, LH, estradiol, and CA 125. The resected ovarian tumor measured 6 cm in diameter, and showed a papillary growth. Histologically, the ovarian tumor was consistent with BET, and the morules with OCN were scattered. Immunohistochemically, OCN were proven to be rich in biotin. An aberrant nuclear expression of beta-catenin was observed in both the tumor cells and the morular cells. Our case may suggest the possibility that the appearance of OCN with or without morules in ovarian tumors is related to endometrioid differentiation of the tumor cells, and should be recognized as a diagnostic clue of ovarian endometrioid tumors. Although female sex hormones have been reported to play a role in the occurrence of OCN, the participation of beta-catenin mutation has also been suggested. PMID- 12588419 TI - Localized malignant mesothelioma of the epithelial type occurring as a primary hepatic neoplasm: a case report with review of the literature. AB - We report a localized malignant mesothelioma of the epithelial type, occurring as a primary hepatic neoplasm in a 64-year-old male. He was found to have a mass located in the right lobe of the liver. Surgery was carried out with resection of the mass from the right hepatic lobe, with partial resection of the diaphragm. Grossly, an ill-defined tumor was present in the hepatic parenchyma. Histologically, the tumor displayed a predominant tubular pattern of growth with a desmoplastic stroma. The tubules were lined by a single layer of cuboidal or flattened cells with pleomorphic vesicular nuclei. A hyaluronidase-digestible, mucin-like substance was demonstrated in the lumen and tumor cytoplasm. The tumor cells were immunohistochemically positive for calretinin, HBME-1, cytokeratin, i.e. AE1/AE3 and CAM 5.2, but negative for carcinoembryonic antigen, CD 34 and Leu M1. Moreover, the tumor cells showed nuclear accumulation of the p53 oncopotein and reacted frequently with Ki-67 antibody. These findings support the concept that malignant mesothelioma of the epithelial type may occur at extrapleural sites. To the best of our knowledge, this is the first reported case of localized malignant primary mesothelioma arising in the liver. PMID- 12588420 TI - Topical application of benzalkonium chloride to the stomach serosa increases gastric emptying time, acid secretion, serum gastrin and size of the mucosa. AB - AIM: In the present study we evaluated the effects of gastric myenteric denervation using benzalkonium chloride (BAC) on the time for gastric emptying, as well as gastric secretion, and mucosal epithelial cell size and population in rats. METHODS AND RESULTS: Wistar rats were treated with topical serosal application of BAC to the stomach. Control animals received saline. Ninety days after surgery, gastric emptying time, gastric acid secretion and serum gastrin levels were studied. Next, the animals were sacrificed and the stomachs were removed, fixed in formalin and histologically processed for histomorphometry of the height, area and volume of the glandular portion, and volume and population of mucous, chief, parietal, G- and labelled cells. BAC animals showed a significant delay in gastric emptying and an increase in gastric acid secretion and serum gastrin levels. These animals also presented a significant reduction of myenteric neuron number, hypertrophy of parietal and chief cells, hyperplasia of G cells and an increase in the gastric mucosa area. CONCLUSION: The absence of the myenteric plexus seems to protect the stomach from the hyperplastic effects of hypergastrinemia. Gastric food stasis may act as a factor triggering morphological and functional alterations of the gastric epithelium. Although gastric food stasis is a common finding in medical practice, its physiopathological consequences are poorly understood and have not been frequently discussed in the literature. PMID- 12588421 TI - Identification of randomly selected colonies of lactobacilli from normal vaginal fluid by pyrosequencing of the 16S rDNA variable V1 and V3 regions. AB - The present study aimed to characterize lactobacilli in vaginal fluid from 23 adult healthy women by using high-throughput DNA sequencing for identification of a large number of randomly selected colonies appearing on Rogosa and blood agar. The typing method was based on broad-range PCR of 16S rRNA gene variable regions V1 and V3, pyrosequencing, and classification of the fragments by alignment with NCBI-catalogued sequences and type strain sequences. Four major groups of sequences were found among the 402 isolates clearly corresponding to Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii when compared to the sequences obtained for type strains. Our results indicate that pyrosequencing of 16S rRNA gene fragments as used here is a fast and reliable method well suited for identification to the species level, even within the Lactobacillus acidophilus complex. PMID- 12588422 TI - Trimethylamine content in vaginal secretion and its relation to bacterial vaginosis. AB - The presence of a fishy odor emanating from women who present with a malodorous vaginal discharge is well known. The odor is due to bacterial reduction of trimethylamine oxide to trimethylamine (TMA) in vaginal secretion. The release of TMA from specimens of vaginal fluid following the addition of alkali is often used in making a clinical diagnosis of bacterial vaginosis (BV). We now report a sensitive gas chromatographic method for analysis and quantification of TMA in vaginal fluid in which weighed samples were used. In addition, a proper diagnosis of BV was obtained using Gram-stained smears of the vaginal fluid according to the method of Nugent et al. (R. P. Nugent et al., J Clin Microbiol 1991;29:297 301). We also diagnosed BV according to Hallen et al. (A. Hallen et al. Genitourin Med 1987;63:386-9). TMA was present in all women with a Nugent score between 7 and 10 and in almost all women diagnosed with BV according to the method of Hallen et al. TMA was not found or was only found in very low concentrations in vaginal fluid from women with Nugent scores of 0 to 3. TMA was also found in four women with a negative sniff test. It seems that high levels of TMA in samples of vaginal fluid are typical for BV regardless of the scoring method used for diagnosis. However, low levels of TMA, <5 microg/g vaginal fluid, do not always correlate with BV. PMID- 12588423 TI - Stomach cancer highly expresses both initiator and effector caspases; an immunohistochemical study. AB - Caspases play an essential role during apoptotic cell death. While caspases 8 and 10 act as initiator caspases of the extrinsic apoptosis pathway, caspase 9 acts as an initiator caspase of the intrinsic apoptosis pathway. Caspase 3 is considered to be the main effector caspase involved in both intrinsic and extrinsic pathways. Alteration of apoptosis is essential for cancer development. Thus, analysis of the expression status of caspases, the main executioners of apoptosis, in cancer tissues is needed for a sophisticated understanding of cancer biology. In the current study, we analyzed the expression of caspases 3, 8, 9 and 10 in 60 advanced gastric adenocarcinomas by immunohistochemistry using a tissue microarray approach. Immunopositivity was observed for caspase 3 in 57 (95%), caspase 8 in 56 (93%), caspase 9 in 54 (90%), and caspase 10 in 58 (97%) of the 60 cancers. While 46 cancers (77%) expressed all of the caspases examined, 14 cancers (23%) showed loss of expression in one or more caspases examined. Normal gastric mucosal cells showed no or weak expression of caspases 3, 8, 9 and 10. Taken together, these results suggest that stomach cancer cells in vivo may need caspase expression for apoptosis. Also, higher expression of the caspases in stomach cancer cells than in normal gastric mucosal cells suggests that apoptosis in susceptible stomach cancer cells might be easily triggered, thereby producing selective pressure to make more apoptosis-resistant cells during tumor development. PMID- 12588424 TI - Public health nursing credentialing and the "pseudo-shortage". PMID- 12588425 TI - Parish nursing: nurturing body, mind, spirit, and community. AB - Parish nursing is a model of nursing care that focuses on health promotion and disease prevention within a faith community. A descriptive study was conducted in the intermountain West to determine how faith communities form parish nursing programs and what their effect is. Thirteen congregations representing eight denominations with parish nurse/health ministries participated. Parish nurses, parish nurse coordinators from two medical centers, pastors, and hospital chaplains (n = 24) were interviewed and provided documents from their programs. Over time, congregations formed parish nursing/health ministries using strategies in a developmental process. The process involved significant support from the pastor, congregation members, and the parish nurses. Collaboration between faith communities and health organizations were successful using a limited domain approach to attain specific health goals. Parish nursing is making a contribution to integrating faith and health practices, promoting health, and increasing accessibility to health care and congregational activities. Conclusions and recommendations are included for future research, practice, and education. PMID- 12588426 TI - The roles of contemporary Mexican American women in domestic health work. AB - Domestic health work is defined as the day-to-day household activities, which are often invisible, that create the backbone of healthy environments and healthy individuals. This article describes the roles of a sample of contemporary Mexican American women in domestic health work. Using an ethnographic design, 13 moderately to highly acculturated women were interviewed to determine their roles in domestic health work. Women's roles fell into two broad categories: being a parent and caring for the family. The findings from this research highlight the burden and conflict of multiple roles in this sample and provide insight into the processes by which contemporary Mexican American women maintain culturally defined roles that they feel are important. It also addresses the burdens that they feel are culturally unnecessary. This research has significance for nurses working in the context of the community and the household in that it explicates women's roles in domestic health work and points to the need for community-based nurses to be aware of the many voices of Mexican American women. PMID- 12588427 TI - Adolescent health promotion scale: development and psychometric testing. AB - The purpose of this study was to assess the psychometric properties of a newly devised instrument, the Adolescent Health Promotion scale (AHP), a 40-item Likert type self-report instrument used to detect unhealthy lifestyles in adolescents. Content validity was considered to be supported based on the findings of previous studies and the observations of a panel of 14 content experts. This study examined the construct validity and reliability of the instrument. The psychometric properties of the AHP, including item analysis, factor analysis, and reliability measures, were assessed based on the responses of 1,128 Taiwanese adolescents. Kaiser-Meyer-Olkin (KMO) measures and Bartlett's sphericity test showed that the samples met the criteria for factor analysis. Factor analysis yielded a six-factor instrument that explained 51.14% of the variance in the 40 items. The six factors were social support, life appreciation, health responsibility, nutritional behaviors, exercise behaviors, and stress management. The Cronbach alpha reliability coefficient for the total scale was 0.932, and alpha coefficients for the subscales ranged from 0.75 to 0.88. The results of this study indicate that the AHP has good construct validity and reliability in Taiwanese and that its use by school health nurses to assess adolescent health promotion programs is warranted. PMID- 12588428 TI - Cultural and noncultural predictors of health outcomes in Korean daughter and daughter-in-law caregivers. AB - In Western cultures, adverse health effects resulting from providing care for impaired elders is well documented for family caregivers, but little is known about the health of Korean caregivers. This study examined the level of depression and physical health of 120 daughter and daughter-in-law caregivers who cared for cognitively or functionally impaired elderly in Korea. It was hypothesized that cultural factors would have a greater effect on caregivers' health outcomes than noncultural factors, but, contrary to this expectation, the effects of noncultural factors were found to outweigh those of cultural factors. Moreover, the caregivers in this study reported a relatively high level of depression, and more caregivers rated their own health as "poor" than did Western caregivers in previous studies. Family caregiving for the impaired elderly is stressful and negatively affects Korean caregivers' health outcomes regardless of societal values such as filial piety and familism regarding parent care in Korea. Culturally acceptable and sensible support programs may be useful in sustaining long-term care at home by Korean daughter and daughter-in-law caregivers. Further family caregiving studies in the Korean sociocultural context are recommended. PMID- 12588429 TI - Adult day care: northern perspectives. AB - This qualitative study explores older adults', caregivers', and nurses' perceptions regarding adult day care (ADC). The study took place in two small towns and one small city in the northern interior region of British Columbia, Canada, with the intent to develop ADC programs that reflect the needs of older persons and their caregivers. The 32 participants contributed their perceptions in focus groups and individual interviews. The interviews, in this descriptive/exploratory study, were audiotaped, transcribed verbatim, and analyzed using inductive, qualitative techniques. The participants acknowledged ADC services as essential to the health and well-being of older adults and their caregivers. The major themes that emerged were need for respite; aging in place; ADC programming; program characteristics; staff knowledge, skills, and attitudes; and northern perspectives. The study participants identified a number of possible reasons for underuse of ADC programming in the north. This study provides information that can facilitate the grounding of ADC policy within the clients' perspectives and a northern context. The participants' thoughts also highlight areas of policy that have broad applicability to the provision of services to the elderly in any setting. PMID- 12588430 TI - Stress and coping in high-risk mothers: difficult life circumstances, psychiatric mental health symptoms, education, and experiences in their families of origin. AB - A pilot study was conducted to test methods and measures used to assess factors related to stress and coping in high-risk mothers and their effect on parenting. Twenty mothers with 12-month-old toddlers were recruited from a WIC population; they came into an observational laboratory where they were interviewed about stressors in their lives and their symptoms of stress and were videotaped interacting with their children. Maternal difficult life circumstances, psychiatric-mental health symptoms, education, maternal experiences in their families of origin, and parenting stress explained 74% of the variance in maternal sensitive-responsiveness with their toddlers in the laboratory setting. The findings support the methods of the study. Clinical implications and implications for future research to assist in the development of interventions for this population are discussed. Increased attention to screening for maternal psychiatric-mental health symptoms and for negative experiences in mothers' families of origin may provide important opportunities for intervention with these mothers. PMID- 12588431 TI - Southern women's response to a walking intervention. AB - The need to change the sedentary habits of many American adults is well recognized. Middle-aged women are an important target group for increased physical activity because of certain health risks such as osteoporosis. In the current study, 31 women between the ages of 30 and 60 from high- and low-income groups (high-income >$50,000; low-income <$50,000 per year) took part in a physical activity intervention. The goal was to increase walking activity to a minimum of 90 min per week. Each woman received 16 telephone calls over a 6-month period in which she was asked to reflect upon the benefits of walking, goal setting, restructuring plans, social support, exercise efficacy, relapse prevention, and maintenance. Content analysis revealed a number of themes emerging from intervention conversations. There were differences between races in walking location and walking partners. Furthermore, there were differences between income groups in beliefs about the benefits of walking and social support. Overall, the intervention appeared to provide a basis for women to develop a walking routine. The women were able to reflect upon their walking routine and attempts to begin a walking routine and to identify how each component of the intervention affected their individual daily routine. PMID- 12588432 TI - Older women and exercise: theory of planned behavior beliefs. AB - Despite well-documented benefits of exercise, aging women remain largely sedentary. Further understanding of beliefs associated with exercise could result in more-effective public health interventions to increase exercise in this vulnerable population. This study examined the relationships between theory of planned behavior constructs and exercise behavior and exercise intention in older women. Constructs from the theory of planned behavior (behavioral beliefs, perceived control beliefs, and normative beliefs) were examined in a sample of 225 women aged 65 and older. Exercise was measured with the Baecke Physical Activity Scale. All women were interviewed, to prevent literacy and vision problems from hampering participation. Significant predictors of exercise behavior were perceived control beliefs and behavioral beliefs. Significant predictors of exercise intentions were perceived control beliefs, behavioral beliefs, and normative beliefs. Specific belief items predicting exercise behavior were that exercise is good for health and that exercise is difficult because of tiredness, as well as the lack of commitment and time. These findings provide partial support for the application of the theory of planned behavior to exercise in older women. The findings suggest that interventions should focus on increasing women's confidence that they can overcome barriers to exercise. PMID- 12588433 TI - Immunohistochemical analysis for histopathological subtypes in pediatric medulloblastomas. AB - Medulloblastomas occurring in children represent a histological spectrum of varying anaplasia and nodularity. In order to determine whether immunohistochemical markers might be useful parameters in subclassifying these tumors, 17 pediatric medulloblastomas, including nine diffuse/non-anaplastic, four diffuse/anaplastic, three nodular/non-anaplastic and one nodular/anaplastic subtypes, were studied. In the present report, we investigate the expression of neural cell adhesion molecule (NCAM), nerve growth factor receptor (NGFR), neurofilament (NF), synaptophysin (SYN), glial fibrillary acidic protein (GFAP), S100, Bcl-2, and Ki-67 by using the immunohistochemistry against specific antibodies. This study showed that NGFR, NF, GFAP and S100 were not detected in anaplastic subtypes of medulloblastomas (0/5), while non-anaplastic subtypes were mainly expressed within the nodules. All 17 tumors were reactive for NCAM, SYN and Bcl-2. In addition, Ki-67 labeling indices for anaplastic subtypes (39.0 +/- 7.42%) were significantly higher than that of non-anaplastic medulloblastomas (11.4 +/- 8.04%; P < 0.0001). These results suggest that immunohistochemical markers are a useful adjunct in characterizing subtypes of pediatric medulloblastomas. PMID- 12588434 TI - Prognostic value of combined analysis of cyclin D1 and estrogen receptor status in breast cancer patients. AB - The amplification of cyclin D1, located on chromosome 11q13, in breast cancer patients has been found to be associated with reduced relapse-free and overall survival; however, there still exists strong controversy about these findings. In order to evaluate the prognostic value of cyclin D1 and other prognostic variables in human breast cancers, we have assessed estrogen receptor (ER) status, cyclin D1, c-erbB2 and p53 overexpression in 175 primary breast carcinomas, and investigated the relationships of prognostic variables to the patient clinical outcome and the association between cyclin D1 overexpression and other prognostic variables. There was some degree of variability in staining intensities and proportions within the same tumor. The overexpression of both cyclin D1 and ER revealed a significantly prolonged survival in univariate analysis (P = 0.020). Among the various prognostic variables, distant metastasis showed a statistically significant association with overall survival. A significant correlation was observed between cyclin D1 overexpression and small size of the primary tumor (P = 0.031), low Bloom and Richardson's histological grade (P = 0.001), and positive ER status (P = 0.000). In contrast to what was previously expected, the present study suggests that the overexpression of cyclin D1 has a tendency to have a positive clinical outcome and a potential role in identifying a subset of patients predicting a good prognosis, particularly when ER is coexpressed. PMID- 12588435 TI - Mucosal high apoptotic activity and low p21WAF1/CIP1 expression and submucosal low proliferative activity in superficially spreading early gastric cancers: comparison with the penetrating growth type. AB - In order to investigate cell kinetics and cell cycle regulator protein expression with reference to the growth pattern of early gastric carcinomas (EGCs), we evaluated a total of 240 EGCs with submucosal invasion clinicopathologically and 106 submucosal invasive lesions immunohistochemically. The incidence of lymph node metastasis was relatively high (36.4%) in the superficially spreading growth (SUP) type tumors whereas the penetrating growth (PEN) type had a low incidence (5.7%, P < 0.001) and correlated with submucosal tumor size. Ki67 labeling was lower in submucosal areas of the SUP-type tumors (median, 37.3%) than the PEN type tumors (51.0%, P < 0.001). ssDNA labeling in the lamina propria, indicative of apoptotic activity, was higher in the SUP-type tumors (0.55%) than in PEN-type (0.30%, P < 0.01) lesions. The expression of cell cycle regulator p21WAF1/CIP1 was lower in the SUP-type tumors (lamina propria 15.6%, submucosa 2.6%) than in PEN-type tumors (lamina propria 26.5%, submucosa 4.4%, P < 0.05-0.001). In conclusion, differences in cell kinetics and p21WAF1/CIP1 expression might influence the growth pattern of EGCs. The SUP-type EGC, characterized by high apoptotic in the lamina propria and low proliferative activities in the submucosa, is associated with frequent lymph node metastasis, suggesting a strong correlation between tumor size in the submucosa and metastatic potential. PMID- 12588436 TI - Combined hepatocellular and cholangiocarcinoma with marked squamous cell carcinoma components arising in non-cirrhotic liver. AB - We report a surgical case of liver tumor, 40 x 35 mm in size, with squamous cell carcinoma (SCC) and hepatocellular carcinoma (HCC) components in a 60-year-old Japanese man with steatohepatitis. Most of the SCC component showed typical intercellular bridge and keratinization, while most of the HCC components showed a thick trabecular pattern with mild to moderate nuclear atypia. Both components transit each other without undifferentiated foci; however, a small foci showing glandular structure was intermediated. No cyst formation was found in the liver. The primary site of the squamous cell carcinoma was not detected in general clinical and radiological examination. Immunohistochemical analysis revealed that part of the HCC components neighboring the SCC showed patchy and weak expression of cytokeratin 7. There are several possibilities for the origin of squamous cell carcinoma in this case: marked squamous metaplastic change of cholangiocarcinoma and/or HCC, and carcinoma originating from pleuripotential stem cells. Irregular fatty changes, scattered giant mitochondria and acellular fibrosis with bridging were seen in the liver; however, this patient had no episode of hepatitis associated viral infection. This is an interesting case of combined hepatocellular and cholangiocarcinoma with marked SCC components arising in a non cirrhotic fibrotic liver. PMID- 12588437 TI - Lipomatous pseudohypertrophy of the pancreas: further evidence of advanced hepatic lesion as the pathogenesis. AB - An 80-year-old Japanese man with liver cirrhosis was suspected of having lipomatous pseudohypertrophy of the pancreas on the basis of results of ultrasonography and a computed tomography scan. He eventually died of hepatic failure. He had no obesity, diabetes mellitus or pancreatic symptoms during his entire clinical course. Autopsy results confirmed lipomatous pseudohypertrophy of the pancreas, cholecystolithiasis, and postnecrotic liver cirrhosis associated with submassive hepatic necrosis was suspected. Although the pathogenesis of lipomatous pseudohypertrophy of the pancreas is not clear, the findings in the present case provide further evidence to support the hypothesis that advanced hepatic lesions cause this lesion. PMID- 12588438 TI - Carcinoid tumor in the female urethral orifice: rare case report and a review of the literature. AB - We experienced an extremely rare tumor in the female urethral orifice in a 57 year-old Japanese woman. To our knowledge, only two cases of primary urethral carcinoid tumor have been reported. The previous reports of urethral carcinoid tumor were recognized in the male middle urethra and penile urethra. The present case was resected, and diagnosed as a carcinoid tumor by histological, immunohistochemical and ultrastructural findings. The tumor cells were stained by chromogranin A, synaptophsin and neuron-specific enolase, and neurosecretory granules were confirmed with electron microscopy. The patient did not complain of any symptoms until 5 years after the resection of the tumor. Therefore, the case we describe here is the first known report of carcinoid tumor in the Japanese female urethra. PMID- 12588439 TI - Primary carcinosarcoma of the vagina. AB - Primary carcinosarcoma of the vagina is a very rare tumor, with only eight cases diagnosed as carcinosarcoma in the literature that we are aware of. We recently encountered a case of primary carcinosarcoma of the vagina in a 75-year-old woman. The patient had a history of hysterectomy and bilateral ovariectomy for uterine corpus cancer at 55 years of age. Recurrence of the cancer was suspected 17 years after the operation and irradiation therapy was performed, but the patient died 3 years after the recurrence. Autopsy revealed a mass lesion in the pelvic cavity that originated in the vagina. Histological examination showed that the tumor contained anaplastic carcinoma, rhabdomyosarcoma, leiomyosarcoma and chondrosarcoma components, and it was diagnosed as carcinosarcoma. The histological diagnosis of the uterine corpus cancer was well-differentiated adenocarcinoma, and there was no sarcomatous component. The carcinosarcoma occurred 17 years after the hysterectomy, and it was concluded to be a primary carcinosarcoma of the vagina. This is the first case of primary vaginal carcinosarcoma in which the epithelial and sarcomatous components were clearly identified histologically and immunohistochemically. PMID- 12588440 TI - Adenomyoma of the small intestine: report of two cases and review of the literature. AB - Adenomyoma of the small intestine is a rare tumor-like lesion composed of exocrine-type ducts and hypertrophic smooth muscle. We describe two cases of adenomyoma of the small intestine. One was an ileal adenomyoma that presented with intussusception occurring in a 7-month-old boy. The other was a jejunal adenomyoma found incidentally in a 63-year-old man with colon cancer. Histologically, the lesions composed of benign ducts and bundles of smooth muscle. The second case was detected on contrast-enhanced computed tomography scan as a small enhancing polypoid mass. We review the previous literature of adenomyoma of the small intestine. PMID- 12588441 TI - Low-grade fibrosarcoma of the proximal humerus. AB - We present the clinical, radiographical and pathological features of low-grade fibrosarcoma of the left proximal humerus in a 23-year-old man in whom it was necessary to distinguish the tumor from desmoplastic fibroma, malignant fibrous histiocytoma and intramedullary well-differentiated osteosarcoma. The patient presented with a 10-day history of pain in his left upper arm sustained when trying to break his fall with his left hand when slipping in the street. Plain radiography revealed an expanding multilobular osteolytic lesion from the proximal metaphysis to the diaphysis of his left humerus, accompanied by a pathological fracture at the distal portion of the lesion. Open biopsy of the lesion was performed twice; however, a conclusive diagnosis could not be obtained. The patient underwent wide excision and prosthetic replacement of the left proximal humerus. Histologically, the resected tumor was composed of both cellular areas and hypocellular areas. Cellular areas revealed a proliferation of bundles of uniform fibroblastic spindle-shaped cells with minimal cellular atypia, mixed with abundant intercellular collagenization. Mitotic figures were occasionally seen. Hypocellular areas showed myxoid features with loose bundles of collagen fibers. The patient demonstrates no evidence of disease 42 months after surgery. It is important to detect the scant atypical cells for the differential diagnosis of low-grade fibrosarcoma and desmoplastic fibroma of bone. PMID- 12588442 TI - Primary sclerosing lipogranuloma with broad necrosis of the scrotum. AB - A-25-year-old man was admitted because of a painless tumor of the scrotum. The patient denied a history of exogenous material injection and trauma in the scrotum. Physical and radiological examination revealed a mass in the scrotum, and blood laboratory tests showed no significant findings except for mild eosinophilia (5.6%). Resection of the mass was performed. The mass was isolated and located in the subcutaneous tissue of the scrotum. The mass was rectangular and symmetrical, and measured 65 x 45 x 15 mm. Histologically, the mass was composed of adipose tissue with fibrosis. Many epithelioid granulomas with multinucleated giant cells of foreign body and Langhans' types and heavy infiltrates of lymphocytes and eosinophils were recognized. Characteristically, the lesion showed broad coagulative and lytic necrosis. Congestion and edema suggestive of ischemia were seen in some areas. Special stains for acid-fast bacteria, gram-positive bacteria and fungi failed to detect any microorganisms. Polymerase chain reaction for mycobacterium tuberculosis revealed no reaction products. Immunohistochemically, the majority of lymphocytes were CD45RO-positive T cells, and S-100 protein-positive cells and CD68-positive macrophages were scattered in small amounts. The appearances were typical for sclerosing lipogranuloma except for the necrosis. Although the pathological mechanism of the broad necrosis is unclear, the necrosis might be the result of ischemia. Our case suggests that primary sclerosing lipogranuloma of the scrotum might show broad necrosis, and that T-cell-mediated immune response might play a part in the formation of lipogranuloma. PMID- 12588443 TI - Rearrangement of immunoglobulin heavy and light chains and VH family in thyroid and salivary gland lymphomas. AB - It is often difficult to differentiate extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) from non-neoplastic inflammatory conditions. Demonstration of clonal lymphoid proliferation by molecular procedures is important for accurate diagnosis. We examined the clonal population of B-cell lymphomas in nine cases of thyroid and two cases of salivary gland B-cell lymphoma using semi-nested polymerase chain reaction (PCR)-based assay for IgH gene arrangement and reverse transcription (RT)-PCR single-strand conformation polymorphism (SSCP) for the detection of IgL gene rearrangement. Clonality was evident in nine out of 11 cases of B-cell lymphomas examined by PCR, and in six of eight cases by RT-PCR SSCP. In addition, analysis of VH families was performed in eight cases. Although VH3 family was frequently used, each case demonstrated the VH4, VH5 or VH6 family. It is possible that the normal counterpart of thyroid or salivary gland lymphoma might be different from peripheral blood B lymphocytes, which usually use VH3 family. Our results indicate that although no clonality was noted in one case by both PCR and SSCP, these molecular methods are useful as supplementary diagnostic tests for both thyroid and salivary gland lymphomas. PMID- 12588444 TI - Chlamydia pneumoniae and multiple infections in the aorta contribute to atherosclerosis. AB - Our previous study on herpesvirus infection including Herpes simplex virus type 1 (HSV-1), Epstein-Barr virus (EBV), cytomegalovirus (CMV) and atherosclerosis revealed that the prevalence of herpesvirus is higher in atherosclerotic aorta than in non-atherosclerotic aorta. Infections with two or three forms of the virus have been found only in atherosclerotic aorta. In our current study, we examined both Chlamydia pneumoniae and Chlamydia trachomatis in herpesvirus infected aortic tissues, by means of immunohistochemistry, polymerase chain reaction, Southern hybridization, in situ hybridization, electron microscopy and electron-microscopic immunohistochemistry. In particular, the bacteria were found in atherosclerotic lesions. In atherosclerotic aorta, 40% of tissues examined were positive for C. pneumoniae in contrast to absence of this bacteria in non atherosclerotic aorta. Elementary bodies of C. pneumoniae were found in macrophage-like cells in the intima of atherosclerotic aorta by electron microscopy. Chlamydia trachomatis was not found in both atherosclerotic and non atherosclerotic aorta. Our findings suggest that multiple infections in aortic tissue contribute to the development of atherosclerosis. Furthermore, the absence of C. pneumoniae compared to herpesviruses in normal arterial tissue suggests that C. pneumoniae is specific for atherosclerotic lesions. In contrast to 'abortive infection' of viruses in arteries, C. pneumoniae infection was demonstrated in macrophages by electron microscopy and electron-microscopic immunohistochemistry in atherosclerotic lesion. Chlamydia pneumoniae may be the most important pathogen related to the development of atherosclerosis. PMID- 12588445 TI - Profile of methylated CpG sites of hMLH1 promoter in primary gastric carcinoma with microsatellite instability. AB - A recent study using colorectal cancer cell lines has identified methylation on a small region of hMLH1 promoter (-248 to -178 relative to the transcription start site) to be critical for gene silencing, but shown that methylation on a more upstream region is frequent in cell lines with hMLH1 expression. Because cultured cell lines have a higher degree of CpG methylation than primary tumors, we attempted to examine methylation profiles of CpG sites of hMLH1 promoter in primary gastric carcinomas with or without microsatellite instability (MSI). Seven cases with MSI and six cases without MSI were assessed for the methylation status of hMLH1 promoter by bisulfite-sequencing. All of the MSI-positive cases previously showed loss of hMLH1 expression and six cases displayed methylated alleles in methylation-specific PCR (MSP) for hMLH1. Sequencing analysis revealed that: (i) CpG sites were overall methylated in MSI-positive tumors with positive MSP results; (ii) a small region (-248-178) was almost invariably methylated in MSI-positive tumors; and (iii) the vast majority of CpG sites were unmethylated in MSI-negative tumors, including a more upstream region (proximal to -248). Our study suggests that methylation of a more upstream region observed in colon cancer cell lines may be an acquired change during cell line establishment and it was not identified in primary gastric carcinomas without MSI. PMID- 12588446 TI - Immunohistochemical localization of endothelial cell markers in solitary fibrous tumor. AB - Solitary fibrous tumor (SFT) is an uncommon tumor first reported in the pleura, but recently described in other tissues. CD34, which is expressed in hematopoietic stem cells, endothelial progenitor cells and vascular endothelial cells, is observed in most SFT and some investigators believe that its expression is a definitive marker of this tumor. In the present study, the expression of vascular endothelial cell markers, such as vascular endothelial growth factor receptor (VEGFR)-1 (flt-1), VEGFR-2 (flk-1/KDR), Tie-2 and c-Met, was examined in SFT to clarify the relationship between SFT and endothelial cells. By immunohistochemical staining of tumor cells from 26 patients, VEGFR-1 was detected in 24 (92%), VEGFR-2 in five (19%), Tie-2 in 14 (54%), and c-Met, a specific receptor of hepatocyte growth factor (HGF) in 23 patients (88%). Furthermore, VEGFR-3 (flt-4) immunoreactivity was detected in eight of 26 patients (31%). In contrast, VEGF, VEGF-C and HGF, which are ligands for the receptors, were not localized in the SFT cells. These findings indicate that most SFT may closely relate to vascular or lymphatic endothelial cells and the endothelial growth factors may contribute to the growth of SFT in a paracrine manner. PMID- 12588447 TI - Malignant phyllodes tumor of the prostate: retrospective review of specimens obtained by sequential transurethral resection. AB - A case of malignant phyllodes tumor of the prostate in a 67-year-old man is reported. The patient was referred to a hospital for urinary retention. From material taken at three transurethral resections of the prostate (TURP), a histological diagnosis of benign prostatic hyperplasia was made. However, at the fourth TURP, phyllodes tumor was diagnosed due to the presence of elongated epithelial ducts and proliferating cellular stroma with mitosis and nuclear atypia. Two months later, total cystoprostatectomy was performed. Histologically, the tumor was composed of dysplastic stromal cells and irregularly elongated epithelial ducts. Five months later the patient developed multiple lung and pelvic lymph node metastases and died. This report documents progression to a higher histological grade of prostatic phyllodes tumor documented with sequential pathological findings obtained from four TURP and surgical specimens over about 3 years. PMID- 12588448 TI - Diffuse mucosal carcinoma of intrahepatic and extrahepatic bile ducts including gallbladder. AB - An autopsy case with a widespread mucosal carcinoma of the biliary tree was reported. A biochemical profile of the bile duct damage was noticed in a woman in her seventies during a gastric examination. Imaging procedures depicted irregular dilatations of intrahepatic bile ducts with a bead-like appearance. Elevated levels of serum alkaline phosphatase and gamma-glutamyltransferase with a negative antimitochondrial antibody persisted. The patient was diagnosed as primary sclerosing cholangitis, she was followed up for 4 years under preservative therapies, and died of anasarca and heart failure. Post-mortem examination showed a diffuse mucosal carcinoma of both intrahepatic and extrahepatic biliary passages including the gallbladder with a minimal invasion and scattered foci of adenoma-like area in part. There was no evidence of gallstones or pre-existing sclerosing cholangitis. The striking features of the tumor were extensive papillary growth, mucus secretion and irregular dilatation of bile ducts. The tumor may bear biological and morphological homology with intraductal papillary mucinous tumor of the pancreas. PMID- 12588449 TI - Malacoplakia of the ovary, fallopian tube and uterus: a case associated with diabetes mellitus. AB - Malacoplakia is a chronic xanthogranulomatous inflammation that most commonly affects the urinary tract and the gastrointestinal system of middle-aged women. It is rarely encountered in a female genital tract, and only a handful of cases of malacoplakia of the ovary have been described. We report an unusual case of malacoplakia extensively involving the ovary, fallopian tube and uterus of a 47 year-old woman with poorly controlled diabetes mellitus. Escherichia coli was cultured from the ovarian lesion. To our knowledge, such an extensive female genital malacoplakia associated with diabetes mellitus has not been reported before. Widespread or atypical site malacoplakia occurring in a patient with systemic disease may result from a diminution of macrophagocytic function, either under the influence of the systemic illness or related to corticosteroid excess. We propose that diabetes mellitus without appropriate medical control may have resulted in impaired leukocyte function which, when combined with E. coli infection, led to the development of extensive malacoplakia in the genital tract of this patient. PMID- 12588450 TI - Unusual case of follicular lymphoma with hyaline-vascular follicles. PMID- 12588452 TI - Oral microbiota associated with hyposalivation of different origins. AB - We analysed and compared the oral microbial flora in four groups with hyposalivation caused by radiation therapy (RT), primary Sjogren's syndrome (pSS), medication or unknown factors (Unknown), or neuroleptic treatment (Neuro). A control group with normal salivary secretion was also included. The subjects included were 54 +/- 8 years old and had 25 +/- 4 teeth. We analysed their microflora in rinsing samples using a cultivation technique. A marked increase in Lactobacillus spp. and Candida albicans was characteristic of the RT group. In the pSS group, 85% of subjects had high numbers of mutans streptococci despite good oral hygiene, frequent dental visits and fluoride use. The Unknown group had an oral flora similar to that of the controls. In the Neuro group, with a stimulated secretion rate similar to that of the Unknown group, the numbers of aciduric and acidogenic microorganisms were close to those in the pSS group. The results indicate that changes in the oral microflora associated with hyposalivation are related to the reason for the hyposalivation rather than to the magnitude of the decrease in the salivary secretion rate. PMID- 12588453 TI - Interleukin-6 and granulocyte-macrophage colony-stimulating factor in apical periodontitis: correlation with clinical and histologic findings of the involved teeth. AB - Apical periodontitis is characterized by the presence of immunocompetent cells producing a wide variety of inflammatory mediators. Releasing cytokines with long range action, such as interleukin-6 (IL-6) and granulocyte-macrophage colony stimulating factor (GM-CSF), apical periodontitis may induce changes in remote organs of the host. This study quantified the levels of IL-6 and GM-CSF in symptomatic and asymptomatic human periradicular lesions. Lesions were also characterized by size and histologic findings. Tissue samples were homogenized and supernatants were assayed using an enzyme-linked immunosorbent assay (ELISA). Correlations between cytokine levels and characteristic features (as single variables) of the lesions were analysed. There was a trend for higher levels of IL-6 and GM-CSF in symptomatic than in asymptomatic lesions, but the difference was not significant. Levels also tended to be higher in large than in small lesions, in polymorphonuclear (PMN) cell-rich than in PMN cell-poor samples, and in epithelialized than in non-epithelialized lesions. Significantly higher levels of IL-6 (778.1 +/- 220.5 pg/microg) and GM-CSF (363.3 +/- 98.4 pg/microg) were found in samples coincidentally possessing symptomatic and epithelialized features than in asymptomatic, small, PMN cell-poor, non-epithelialized lesions (IL-6: 45.2 +/- 13.1 pg/microg and GM-CSF: 135.1 +/- 26.4 pg/microg). These results suggest that symptomatic lesions containing epithelial cells represent an immunologically active stage of apical periodontitis, whereas asymptomatic, small, PMN cell-poor, non-epithelialized lesions represent healing apical lesions. PMID- 12588454 TI - Contrasting responses of human gingival and periodontal ligament fibroblasts to bacterial cell-surface components through the CD14/Toll-like receptor system. AB - We compared human periodontal ligament fibroblasts with human gingival fibroblasts isolated from the same donor to examine interleukin-8 (IL-8) responses of the cells to Salmonella lipopolysaccharide, a water-soluble peptidoglycan from Staphylococcus epidermidis and the synthetic muramyldipeptide, with special reference to the possible involvement of the CD14/Toll-like receptor (TLR) system of the cells in the responses. Human gingival fibroblasts expressed CD14 on their surfaces and strongly expressed CD14 mRNA, while human periodontal ligament fibroblasts showed considerably lower levels of expression in both respects. Both cells expressed mRNA of TLR-related molecules, i.e. TLR2, TLR4, MD 2 and MyD88, although human periodontal ligament fibroblasts expressed TLR2 more strongly than human gingival fibroblasts. Human gingival fibroblasts exhibited a stronger IL-8 response than human periodontal ligament fibroblasts to lipopolysaccharide, while human periodontal ligament fibroblasts exhibited a response comparable to, or slightly stronger than, that of human gingival fibroblasts to S. epidermidis peptidoglycan and muramyldipeptide. The IL-8 responses of both cells to lipopolysaccharide and S. epidermidis peptidoglycan were completely inhibited by antihuman CD14 monoclonal antibody (MAb). The responses of both cells to lipopolysaccaride were significantly inhibited by antihuman TLR4 MAb, while those to S. epidermidis peptidoglycan were inhibited by antihuman TLR2 MAb. In contrast, muramyldipeptide activated both types of cells in a TLR2- and TLR4-independent manner, although the activities of muramyldipeptide on human gingival fibroblasts, but not human periodontal ligament fibroblasts, were significantly inhibited by anti-CD14 MAb. PMID- 12588455 TI - Tracing genotypes of mutans streptococci on tooth sites by random amplified polymorphic DNA (RAPD) analysis. AB - The aim of this study was to clarify the distribution and persistence of mutans streptococci on different tooth sites in the same oral cavity. Thirteen subjects, aged 20-40 years, were examined. Salivary levels of mutans streptococci, caries prevalence, oral hygiene habits and status of tooth surfaces sampled were recorded. Plaque samples were obtained from four sites, the mesial and buccal surfaces of the first permanent molar on the right side of the lower jaw (46m and 46b), the distal surface of the first permanent premolar (24d) and the mesial surface of the second permanent premolar (25m) on the left side of the upper jaw, using sterile toothpicks on two occasions at 4-7-month intervals. The samples were cultivated on site-specific Strip mutans. Up to 10 colonies/site were isolated when present and genotyped by random amplified polymorphic DNA (RAPD) analysis, after species identification with PCR. Genotyping was also performed by restriction endonuclease analysis (REA) on 148 isolates, and results were consistent with the RAPD results. Most mutans streptococcus-positive samples were obtained from 46m. Within each individual, the same genotype occurred on at least two sites on all but one sampling occasion. A maximum of seven different genotypes were found in an individual. For a particular tooth site, four genotypes occurred simultaneously and taking both sampling occasions together the maximum was six different types. The same genotypes/types were found again after 4-7 months on 25 sites in 12 subjects. Fifteen sites were mutans streptococcus positive on only one sampling occasion. The results indicate that several different genotypes of mutans streptococci colonize a tooth site, and that the same genotype colonizes several sites in the same oral cavity. Persistence of genotypes on a site for several months and interindividual differences in the occurrence of genotypes were also found. PMID- 12588456 TI - Porphyromonas gingivalis antigen preferentially stimulates T cells to express IL 17 but not receptor activator of NF-kappaB ligand in vitro. AB - Although T cells have been implicated in the pathogenesis and are considered to be central to both their progression and control of chronic inflammatory periodontal diseases, the precise contribution of T cells to tissue destruction has not been fully clarified. Recently, interleukin (IL)-17 and receptor activator of Nuclear factor kappaB NF-kappaB ligand (RANKL) have received much attention as a result of their proinflammatory and bone metabolic roles, respectively. We therefore investigated the effect of outer membrane protein (OMP) from Porphyromonas gingivalis (P. gingivalis) on the expression of IL-17 and RANKL in peripheral blood mononuclear cells (PBMCs) and compared these between gingivitis and periodontitis, which are representative of stable and progressive lesions, respectively. The in situ expression of these molecules was also examined. P. gingivalis OMP stimulated PBMCs to express IL-17 at both the mRNA and protein level. Although the mean expression of mRNA was not different between the two groups, the mean level of IL-17 in the culture supernatants was higher in gingivitis patients than in periodontitis patients. However, the frequency of IL-17-positive samples was higher in the periodontitis patients. This stimulatory effect was not evident for RANKL expression in either periodontitis or gingivitis patients. In gingival tissue samples, IL-17 mRNA was detected in gingivitis more frequently than in periodontitis. The expression of RANKL mRNA was much lower than that of IL-17 in terms of both level and frequency. These results suggest that IL-17 but not RANKL may be involved in the pathogenesis of periodontal diseases. However, there may be negative regulatory mechanisms for IL-17 in gingivitis. PMID- 12588457 TI - Subgingival microbiota of renal transplant recipients. AB - Renal transplant patients undergoing immunosuppressive therapy may experience periodontal side-effects such as gingival overgrowth. This study evaluated the subgingival microbiota of renal transplant recipients with or without periodontal tissue destruction who may have concurrent gingival enlargement. Subgingival paper point samples taken from the deepest probing sites of 38 subjects (one per patient) were examined using direct microscopy and culture techniques. A complex microflora comprising gram-positive and gram-negative cocci, rods and filaments, fusiforms, curved rods and spirochetes was observed using microscopy. Yeasts were occasionally detected. Significantly higher proportions of gram-positive morphotypes, including gram-positive cocci, were observed in samples from periodontally healthy patients. The predominant cultivable microflora from anaerobic culture comprised several species of facultative and obligate anaerobes. Colonization of the subgingival sites by 'foreign' microbes that are normally dermal, intestinal or vaginal flora was detected in up to 50% of the samples. High mean proportions of lost or unidentified species were also occasionally noted. The results showed that the subgingival biofilm of renal transplant recipients with chronic periodontitis comprised mainly gram-negative rods and spirochetes. Besides the usual predominant cultivable subgingival microbiota associated with periodontitis, the high prevalence of unidentified and 'foreign' microbes indicates the possibility of subgingival microbial alteration in renal transplant patients. PMID- 12588458 TI - Characterization of in vitro oral bacterial biofilms by traditional and molecular methods. AB - The aim of this study was to compare culture-based bacterial isolation methods with direct amplification and cloning of 16S rRNA genes from oral biofilms grown in an in vitro model. The model used was a constant depth film fermentor which was inoculated with pooled human saliva. The use of culture techniques and cloning resulted in the identification of 36 different bacterial species from the saliva inoculum and from the biofilms. Of these, only five were detected solely by molecular methods. Three taxa were detected which, according to the databases, were unidentified. Using the molecular methods of detection, differences in the number of species observed were found using different 16S rRNA gene primers and numbers of PCR cycles. We have shown that microcosm supragingival plaque biofilms grown in a fermentor consisted of a community most of the members of which could be cultivated on laboratory media. PMID- 12588459 TI - Comparison of different techniques of quantitative PCR for determination of Streptococcus mutans counts in saliva samples. AB - Saliva samples from 16 children with current caries activity were investigated for Streptococcus mutans using three different PCR techniques, and the results were compared with those of selective cultivation on mitis salivarius agar with bacitracin (MSB) (I, II: LightCycler - competitive PCR end-point analysis; III: LightCycler - kinetic real-time analysis; IV, V: block cycler - competitive PCR end-point analysis; VI: cultivation on MSB agar). In groups I, III, IV and VI the saliva samples were analyzed directly. A DNA preparation before PCR with added competitors was carried out in groups II and V to exclude the influence of PCR inhibitors. The coefficients of correlation ranged from 0.97 to 0.98 among the competitive PCR methods, 0.8 to 0.85 for competitive vs. real-time PCR and 0.5 to 0.65 for PCR vs. cultivation methods. Competitive PCR on the real-time instrument was found to be more rapid than, comparably sensitive to, but less reproducible than competitive PCR on a block cycler. PMID- 12588460 TI - Immunohistochemical localization of Toll-like receptors 2 and 4 in gingival tissue from patients with periodontitis. AB - The present study investigated the expression of Toll-like receptor (TLR) 2, TLR4, cluster of differentiation (CD) 14 and CD1a in human periodontitis gingiva using immunohistochemical methods. The specimens were classified according to the degree of inflammation into three groups (mild, moderate and severe). We established three zones in which to evaluate the ratios of TLR2-, TLR4-, CD14- and CD1a-positive cells to total cells in the connective tissues of each section. TLR2 and TLR4 were expressed in human periodontal tissues, and the ratio of TLR2 positive cells was highest overall in zone 1 (connective tissue subjacent to pocket epithelium) of the severe group and that of TLR4-positive cells was higher in the severe group than in the other groups. These results suggest that TLR2 and TLR4 participate in the innate immune response to stimulation by bacterial products in periodontal tissues. The ratio of CD14-positive cells was lowest overall in zone 1 of the severe group and that of CD1a was higher in the severe group than in the other groups. These results suggest that CD14 may be down regulated during the development of inflammation and/or dendritic cells might infiltrate chronically inflamed gingival tissue. PMID- 12588461 TI - Denaturing gradient gel electrophoresis analysis to study bacterial community structure in pockets of periodontitis patients. AB - Bacteria are involved in the onset and progression of periodontitis. A promising molecular technique, denaturing gradient gel electrophoresis (DGGE), to study microbial population dynamics in the subgingival pocket is presented. Twenty three samples were taken from the subgingival pockets of nine patients and six healthy family members. From four periodontitis patients, 12 samples were evaluated before, 1 day after and 3 months after treatment. Part of the 16S rRNA gene of all bacteria was amplified by PCR and separated by DGGE, creating banding patterns representative of the community structure. Shifts in composition and diversity of the microbial population could be determined semiquantitatively, and this showed that treatment resulted in a decrease in the diversity of the population. After 3 months a microbial population 33-47% different from the population before treatment had re-established. Intense bands representing Exiguobacterium aurantiacum were present in 13 out of 25 samples, indicating that this species may play a role in periodontal disease. PMID- 12588463 TI - The effect of glucagon-induced gastric relaxation on TLOSR frequency. AB - This study aimed to determine the effect of glucagon-induced gastric relaxation on the frequency of transient lower oesophageal sphincter relaxations (TLOSRs). Eight normal subjects (four male, age 18-52 y) were studied after a 6-h fast using a combined manometric barostat assembly. The recording was divided into two 1-h sessions: (1) a baseline period with the barostat set at minimal distending pressure (MDP) + 2 mmHg and (2) a period with continuous glucagon or placebo infusion with barostat set at MDP + 2 mmHg. Patients were studied on two different days and randomly received glucagon (4.8 microg kg(-1) bolus followed by 9.6 microg kg(-1) h(-1) infusion) on 1 day and placebo (saline) on another. Lower oesophageal sphincter (LOS) pressure, frequency of TLOSRs, and barostat bag volumes were determined for both placebo and glucagon infusion. Glucagon induced significant fundal relaxation compared with placebo (P < 0.05) and significantly decreased baseline LEOS pressure (P < 0.05). The frequency of TLOSRs was not altered by glucagon infusion compared with placebo. Despite causing substantial proximal stomach relaxation, glucagon did not increase TLOSR frequency. This suggests that the relevant gastric mechanoreceptors responsible for triggering TLOSRs do not respond to passive elongation. PMID- 12588464 TI - Effect of the 5-HT1 agonist sumatriptan on oesophageal motor pattern in patients with ineffective oesophageal motility. AB - The 5-HT1 agonist sumatriptan (SUM) elicits an increase in amplitude of oesophageal motor waves and of lower oesophageal sphincter (LOS) tone in healthy subjects. The aim of the study was to evaluate whether such an effect occurs also in patients with ineffective oesophageal motility (IOM). 16 patients (nine males and seven females, age range 34-55 years) with chest pain and mild to moderate dysphagia were studied; all had undergone previous cardiologic, radiologic and upper gastrointestinal endoscopic exams that were normal. An oesophageal manometry was performed using an electronic probe to record swallows, oesophageal, LOS and gastric motility. The patients whose motor pattern were compatible with IOM (>30% of motor waves with amplitude <30 mmHg and/or non transmitted) received SUM or placebo 6 mg s.c., injected in the morning and in the afternoon in a random order. The data analysis was limited to 1 h before and 1 h after the drug injections. Ten out of the 16 patients showed an IOM motor pattern. The administration of SUM caused a significant increase in the number of swallows (SUM 99.5 +/- 15.4 vs 78.6 +/- 16.1 basal, P = 0.03) and of primary oesophageal motor waves (SUM 89.6 +/- 13.4 vs 67.2 +/- 12.9 basal, P = 0.04) with no significant changes in the percentage of swallows associated with propagation. Placebo was not associated with increase in the number of swallows (80.3 +/- 14.6, P = 0.9) or of primary oesophageal motor waves (70.1 +/- 12.3, P = 0.7). The amplitude and the percentage of propagated oesophageal motor waves as well as the mean basal LOS tone were unaltered by SUM. There was no change in the symptoms reported after SUM. Although effective in healthy subjects, SUM 6 mg s.c. improves only the numbers but not the amplitude or propagation of oesophageal motility of patients with IOM. The 5-HT1 pathway and its acute stimulation seem to play only a minor role in the pathogenesis of such a disease. PMID- 12588465 TI - Gastric electrical stimulation at proximal stomach induces gastric relaxation in dogs. AB - Gastric electrical stimulation (GES) improves symptoms in patients with gastroparesis. However, the underlying mechanisms remain unclear. To determine if GES at proximal and distal stomach could affect the biomechanical properties of the stomach, thus contributing to the beneficial effect of GES. Four pairs of electrodes were implanted along the greater curvature of the stomach in seven dogs. Gastric tone and compliance was assessed with a barostat. Measurements were obtained randomly during control and proximal and distal stimulation (4 mA, 375 ms and 6/18 cpm). Data as mean or median (25-75th percentiles). Gastric compliance was not affected by proximal and distal GES. Gastric tone was significantly reduced during proximal GES: 82.0 (66.8, 89.1) mL vs control 49.7 (39.6,75.9) mL at 6 cpm (P = 0.016), and 90.6 (54.5, 117.9) mL vs control 62.8 (39.6, 75.9) mL at 18 cpm (P = 0.031). Tone was not affected by distal GES at 6 cpm: 95.8 (46.3, 106.7) mL vs control 75.2 (49.7, 86.1) mL (P = 0.47) and at 18 cpm: 80.4 (38.1, 170.3) mL vs control 62.8 (44.6, 156.3) mL (P = 0.44). Proximal GES induces gastric relaxation. This effect, if seen also in humans, may explain, in part, the symptomatic improvement associated with GES therapy in patients with gastroparesis. PMID- 12588466 TI - Colorectal transport during defecation in patients with lesions of the sacral spinal cord. AB - Normal defecation involves reflexes between the colorectum and sacral spinal cord. Lesions of the conus medullaris or cauda equina interrupt such reflex arches and cause constipation. The aim of the study was to compare colorectal transport during defecation in patients with sacral spinal cord lesions and healthy volunteers. Ten patients with sacral spinal cord lesions (six men and four women, age 21-57 y, median = 36) and 16 healthy volunteers (10 men and six women, age 22-42 y, median = 30) took one or two doses of 111In-labelled polystyrene pellets perorally to mark colorectal contents. Abdominal scintigraphy was performed before and after defecation. Total colorectal emptying and segmental antegrade or retrograde transport was computed. Median colorectal emptying during normal defecation was 81% of the rectosigmoid (range: 53% of the rectosigmoid to complete emptying of the rectosigmoid and 40% of the descending colon) in healthy volunteers and 27% of the rectosigmoid (range: 0-44% of the rectosigmoid) in patients with conal/cauda equina lesions (P < 0.001). Median antegrade transport was 82% (control group) vs 27% (patients) of the rectosigmoid (P < 0.001), 38% vs 4% of the descending colon (P < 0.02), 13% vs 1% of the transverse colon (P = 0.28), and 4% vs 2% of the caecum/ascending colon (P = 0.76). It is concluded that damage to reflex arches between the colorectum and the sacral spinal cord significantly reduces emptying of the rectosigmoid and descending colon during defecation. PMID- 12588467 TI - Capsaicin-sensitive extrinsic afferents are involved in acid-induced activation of distinct myenteric neurons in the rat stomach. AB - Challenge of the rat gastric mucosa with 0.5 mol L(-1) HCl activates nitrergic neurons in the myenteric plexus as visualized by c-Fos immunohistochemistry. In the present study, we characterized the activated neurons more extensively by their chemical coding and investigated whether a neural pathway that involves capsaicin-sensitive extrinsic afferents and/or cholinergic neurons transmitting via nicotinic receptors contributes to the activation of myenteric neurons. In multiple labelling experiments, c-Fos was examined for co-localization with nitric oxide synthase (NOS), vasoactive intestinal peptide (VIP), neuropeptide Y (NPY), enkephalin (ENK), gastrin-releasing peptide (GRP), substance P (SP), calbindin D-28k (CALB) and neurofilament 145 (NF 145). All c-Fos-positive neurons were immunoreactive for NOS, VIP, NPY and NF 145, but not for SP, ENK, GRP and CALB. Nerve fibres co-expressing NOS, VIP and NPY were predominantly found in the external muscle layer and in the muscularis mucosae but rarely in the mucosa. Pre treatment with capsaicin or hexamethonium or a combination of both pre-treatments reduced HCl-induced c-Fos expression by 54, 66 and 63%, respectively. Acid challenge of the stomach, therefore, leads to activation of presumably inhibitory motor neurons responsible for muscle relaxation. Activation of these neurons is partly mediated by capsaicin-sensitive afferents and involves ganglionic transmission via nicotinic receptors. PMID- 12588468 TI - Acid challenge delays gastric pressure adaptation, blocks gastric emptying and stimulates gastric fluid secretion in the rat. AB - Functional dyspepsia can be associated with impaired gastric relaxation in response to food intake and delayed gastric emptying. In this study, we investigated whether luminal hydrochloric acid (HCl) may reproduce these motor alterations in phenobarbital-anaesthetized rats via activation of extrinsic neural pathways. Intragastric pressure (IGP) changes induced by a 2-mL fluid bolus were recorded with an oesophageal catheter, and gastric emptying was determined via the fluid volume recovered from the stomach 30-min post-bolus. Experiments involving acute nerve transections or pharmacological blockade of nitric oxide synthesis revealed that the initial increase of IGP after a 0.35 mol L(-1) HCl bolus is dampened by duodenogastric and gastrogastric relaxation reflexes depending on vagal and splanchnic pathways as well as nitric oxide. Compared with saline, HCl (0.15-0.5 mol L(-1)) delayed the subsequent decrease (adaptation) of IGP, inhibited gastric emptying and stimulated gastric fluid secretion as seen in stomachs with ligated pylorus. The acid-evoked delay in IGP adaptation and inhibition of gastric emptying involved duodenogastric and duodenopyloric extrinsic nerve reflexes, whereas the gastric fluid secretion was independent of the extrinsic innervation. It is proposed that the gastropyloric motor changes induced by luminal acid challenge have a bearing on the motor disturbances underlying functional dyspepsia. PMID- 12588469 TI - Effect of successive swallows on oesophageal motility of normal volunteers, patients with Chagas' disease and patients with idiopathic achalasia. AB - Most frequently, ten swallows of a 5-mL bolus of water are performed during oesophageal manometry. Our hypothesis is that five swallows may produce the same results. We studied the oesophageal contraction parameters of 40 volunteers, 75 patients with Chagas' disease and 14 patients with idiopathic achalasia. Motility was recorded at 5, 10 and 15 cm above the lower oesophageal sphincter. The subjects performed ten swallows of a 5-mL bolus of water alternated with ten dry swallows with an interval of at least 30 s. We measured the amplitude, duration, peristaltic velocity, number of failed and number of simultaneous contractions of the initial five and final five dry and wet swallows. The comparison of dry and wet swallows showed the differences already known. The comparison of the parameters of the initial five swallows with those of the final five swallows showed no differences. Thus, when the initial five or the final five swallows were considered, there was no change in the conclusions reached by the comparison of patients and volunteers and of dry and wet swallows. We conclude that five swallows may be sufficient for the manometric examination of oesophageal parameters in Chagas' disease and idiopathic achalasia. PMID- 12588470 TI - Effects of position on oesophageal function: studies using combined manometry and multichannel intraluminal impedance. AB - Multichannel intraluminal impedance (MII) allows assessment of intraoesophageal bolus transit. In the supine position, bolus transit is produced almost exclusively by peristaltic contractions; in the upright position, gravity also contributes to bolus transit. MII and peristaltic pressures were measured in four positions (0, 30, 60 and 90 degrees ) using ten swallows (5 cc each) of both water and viscous liquid with body position determined by random choice. Tracings were analysed for total bolus transit time: time interval between bolus entry at 20 cm above and bolus exit at 5 cm above the lower oesophageal sphincter (LOS) and contraction amplitudes at 5 and 10 cm above the LOS. Statistical comparison of mean values of all four body positions was done using anova and Bonnferoni post-test. Ten normal subjects (five females and five males, age 24-45 years) completed the study. At each body position, liquid material transited faster (P < 0.001) than viscous material. Both liquid and viscous materials transited at lower inclinations (0 and 30 degrees ) significantly slower than at higher inclinations (60 and 90 degrees ). There was an almost perfect inverse linear correlation between angle of inclination and bolus transit time for both liquid (r = -0.99) and viscous (r = -1.00) boluses (Spearman correlation r > 0.99 and P < 0.02 for both substances). Contraction amplitudes for liquid vs viscous material were not significantly different at a given degree of inclination. Mean distal oesophageal amplitude declined with increasing inclination. Combined MII OM identifies and quantifies the effects of gravity on the dichotomy between specific pressures measured by OM and function assessed as transit measured by MII. PMID- 12588471 TI - Modulation of human swallowing behaviour by thermal and chemical stimulation in health and after brain injury. AB - Few data support thermal or chemical stimulation as therapy for neurogenic dysphagia. Our aims were to explore the behavioural effects of thermal (cold) and chemical (citrus) modalities on water swallowing in health (n = 65, mean age 45 years, 44 females) and acute stroke (n = 22, mean age 67 years, eight females). Multiple randomized timed 50-mL swallowing tests were performed for each of four water conditions: (a) room temperature (RT), (b) cold (CD), (c) citrus (CT) and (d) combined cold and citrus (CD + CT). The inter-swallow interval (ISI), swallowing volume velocity (speed), and volume per swallow (capacity) were measured. In health, compared to RT, only CD + CT slowed the speed (12.3 +/- 0.5 vs 10.3 +/- 0.5 mL s(-1), P < 0.03) and decreased the capacity (16.4 +/- 0.9 vs 14.6 +/- 0.7 mL per swallow, P < 0.02) of swallowing. ISI was unaffected, except by CD + CT in healthy young subjects (<60 years) where it was reduced (1.44 +/- 0.02 vs 1.30 +/- 0.02 s, P < 0.02). Despite smaller volumes ingested by stroke patients, CD + CT, compared to RT, again slowed both the speed (3.8 +/- 0.4 vs 4.5 +/- 0.5 mL s(-1), P < 0.03) and capacity (7.6 +/- 0.7 vs 8.5 +/- 0.7 mL per swallow, P < 0.03) of swallowing but had no effect on ISI. We conclude that combined thermal and chemical modification of water consistently alters swallowing behaviour in health and after cerebral injury. These findings have relevance in the management of neurogenic swallowing problems. PMID- 12588472 TI - Efficacy of alosetron in irritable bowel syndrome: a meta-analysis of randomized controlled trials. AB - The 5HT3 receptor antagonist alosetron has been tested in several trials on irritable bowel syndrome (IBS) patients. The aim of the present meta-analysis was to determine its effect on adequate relief of pain or global improvement of symptoms in IBS patients. Six large, multicentre, randomized, placebo-controlled trials fulfilled pre-set criteria for high quality and were included in the meta analysis; 1762 patients were randomized to alosetron treatment and 1356 to placebo. Seventy-five per cent of the patients experienced diarrhoea-predominant IBS and 93% were females. The pooled odds ratio for adequate relief of pain or global symptoms improvement was 1.81 [95% confidence interval (CI) 1.57-2.10). The average number of patients needed to treat with alosetron for one patient to achieve improvement over placebo treatment was seven (95% CI 5.74-9.43). The present analysis shows that alosetron 1 mg b.i.d. positively impacts global symptoms, and pain and discomfort in non-constipated IBS female patients. One in four patients treated with alosetron may develop constipation. The efficacy of alosetron is unclear in male patients. PMID- 12588474 TI - Total quality management in dermato-mycology in Germany. AB - Mycoses of the skin are extremely frequent. The clinical presentation often resembles the one of other frequent inflammatory skin diseases not caused by fungi. Adequate diagnostics are critical for the establishment of proper treatment. A total quality management system has been devised in Germany during the last 5 years. There are several guidelines addressing various aspects of the management of various types of disease as well as structure quality of laboratory diagnostics. Continuous medical education and so-called ring trials are major aspects of implementation. PMID- 12588476 TI - Identification of clinically relevant Trichosporon species. AB - A dichotomous identification key to pathogenic species of the basisiomycetous genus Trichosporon Behrend is provided. It is based on growth tests with carbon sources not traditionally used in yeast taxonomy, viz. uric acid, ethylamine, L-4 hydroxyproline, tyramine and L-phenylalanine as sources of carbon and nitrogen, and polygalacturonate, quinate, 4-ethylphenol, 2,3-dihydroxybenzoate and orcinol as sole carbon sources. Of the standard growth tests, assimilation of L-rhamnose and the maximum growth temperature proved to be useful. In addition to medically relevant species, other species able to grow at 37 degrees C were treated as well. PMID- 12588477 TI - Mycological and histological investigations in humans with middle ear infections. AB - The aim of our investigations was to characterize fungal colonization of the ear in immunocompetent patients. From 1993 to 2000, 128 patients supposed to suffer from otomycosis were included. Mycological examination conducted by direct microscopy and fungal cultures was performed on 139 specimens. Among these, 115 patients suffered from chronic otitis media with persisting tympanum perforation and otorrhea. A further 13 patients had clinical signs of an otitis externa only. Out of 139 samples, fungi were identified in the auditory canal (n = 54), on the tympanic membrane (n = 5), and in the middle ear (n = 5). Two-thirds were as moulds and one-third yeasts. The dominating species were Aspergillus niger and Candida parapsilosis. Samples from 15 patients supposed to have mastoiditis or cholesteatoma were examined histologically. Fungal hyphae were observed in the middle ear cavity and/or between horny lamellae of cholesteatoma in four patients. In the middle ear of immunocompetent patients chronic-hyperplastic (polypoid) inflammation was detected with increased production of mucus, which probably promotes colonization by pathogenic fungi in the middle ear as well as in the auditory canal. Invasive fungal growth into the subepithelial connective tissue was not observed. PMID- 12588478 TI - Prevalence of Aspergillus fumigatus and other fungal species in the sputum of adult patients with cystic fibrosis. AB - Aspergillus fumigatus is often found in the respiratory tract secretions of patients with cystic fibrosis (CF), although the role of the fungus for progression of pulmonary disease remains unclear. This study aimed to investigate the frequency of A. fumigatus and other fungi in sputum of adult CF patients using different methods for culture and microscopy. Results from the analysis of 369 samples from 94 patients showed that A. fumigatus could be isolated in 45.7% of patients. Other moulds were rare, but the yeast Candida albicans was another frequent isolate, detected in 75.5% of patients. A comparison of different culture media showed no difference between a selective medium developed to specifically inhibit Pseudomonas aeruginosa and a standard fungal culture medium for growth of A. fumigatus, although both were more efficient for detection of fungi than other bacterial culture media. Fluorescent microscopy with calcofluor white was more sensitive for detection of fungal hyphae in undiluted sputum than standard methylene blue staining. This study shows that A. fumigatus and C. albicans have a high frequency in adult CF patients. Microbiological analysis should routinely include methods for specific identification of fungi to monitor for potential complications arising from fungal disease in these patients. PMID- 12588479 TI - Clinical study on liposomal amphotericin B (Ambisome) in deep fungal infections in China. AB - Ambisome (L-Amb) was used to treat nine cases of meningitis or menigoencephalitis by Cryptococcus neoformans and 28 cases of other deep fungal infections. A retrospective study on conventional amphotericin B (C-Amb) was performed as the control. A series of indices was observed including curative effect, fungal clearance rate, course of treatment, daily dose, cumulative dose and adverse effects. Nine cases of cryptococcal meningitis or menigoencephalitis treated with Ambisome were clinically cured with an effective rate of 100%, within a mean course of 50 days, which was shorter than that of C-Amb, by a mean cumulative dose of 1807.2 mg, which was not statistically significant in comparison with C Amb. Fungal clearance rate on the second month of treatment was 89% with Ambisome, which was higher than that of C-Amb. Twenty-eight cases of other deep fungal infections treated with Ambisome were clinically cured with an effective rate of 92%, within a mean course of 19.3 days, by a mean cumulative dose of 907.5 mg, and fungal clearance rate on the second and third month was 75 and 92%, respectively. The adverse effects by Ambisome decreased evidently compared with those by C-Amb. PMID- 12588480 TI - Inhibition of human pathogenic fungi by ethnobotanically selected plant extracts. AB - In this study, 36 extracts derived from 29 plant species selected using an ethnobotanical approach were tested for antifungal activity against a taxonomically diverse group of 13 human pathogenic fungi. We compared the inhibitory characteristics of these plant extracts with those of the commonly used antifungals, amphotericin B and ketoconazole, and the plant-derived antifungal, berberine. Several plant extracts, notably those from Zingiber officinale (ginger) and Juglans cinerea (butternut), had pronounced antifungal activity against a wide variety of fungi, including strains that were highly resistant to amphotericin B and ketoconazole. Further exploration of Z. officinale as an antifungal is warranted as this species is generally regarded as safe for human consumption. PMID- 12588481 TI - Tinea pedis observed in Cagliari, Italy, between 1996 and 2000. AB - The aim of this study was to verify the incidence of tinea pedis in patients observed in the Department of Dermatology of the University of Cagliari, Italy, in the period from 1996 to 2000. We examined 722 patients, 536 with lesions and 186 without lesions of the feet and in 169 of them (23.4%) we diagnosed tinea pedis. All patients suffering from tinea pedis belonged to the group with lesions. In the 536 patients with clinical manifestations which were evident to a certain degree, microscope and cultural examination gave positive results for dermatophytes in 169 cases (31.5%). The most frequently isolated dermatophyte was Trichophyton mentagrophytes (51.5%), followed by Trichophyton rubrum (45.2%) and Epidermophyton floccosum (3.3%). In the 186 patients without lesions, direct microscope examination was consistently negative while cultural examination showed the growth of sparse colonies of Candida albicans in two cases (1.1%) We analysed distribution by sex, age, residence, occupation and clinical manifestations. Stressing the high frequency of tinea pedis in this region, the discovery of a dermatophytic infection of the feet with an absence of signs and symptoms is an exceptional event. PMID- 12588482 TI - Relapse of coccidioidomycosis despite immune reconstitution after fluconazole secondary prophylaxis in a patient with AIDS. AB - We report a case of relapse of coccidioidomycosis in an HIV-infected person who discontinued maintenance azole therapy following a response to antiretroviral therapy. This experience raises concern about the safety of withholding secondary prophylaxis for coccidioidomycosis in persons with HIV infection who have achieved immunological responses to antiretroviral therapy. PMID- 12588483 TI - Paecilomyces variotii central nervous system infection in a patient with cancer. AB - Paecilomyces variotii was isolated from two subsequent cerebrospinal fluid (CSF) specimens of a cancer patient. Identification was confirmed through beta-tubulin and rDNA ITS sequencing. MICs were determined for seven antifungal agents; the isolate was found to be susceptible to amphotericin B (AMB), itraconazole (ITZ), ketaconazole (KTZ) and 5-fluorocytosine (5FC) but resistant to fluconazole (FLZ) and miconazole (MCZ). Despite antimycotic therapy, the infection proved to be fatal. PMID- 12588484 TI - Pseudomembranous tracheobronchial aspergillosis: a rare manifestation of invasive aspergillosis in a non-neutropenic patient with Hodgkin's disease. AB - Pseudomembranous tracheobronchial aspergillosis coincident with systemic pulmonary aspergillosis represents a rare manifestation of fungal infection in immunocompromized hosts. We report on a patient with recurrent Hodgkin's disease, showing this infectious pattern after treatment with corticosteroids within the antineoplastic schedule, whereas neutropenia--the main risk factor for mold infections--had not occurred. An impaired number of helper T lymphocytes was merely detected as an additional, but hypothetical risk factor, when investigating the status of immunosuppression. Treated systemically with amphotericin B, the patient recovered quickly, although reported mortality rates are disastrous. What is crucial for the clinical management is an early diagnosis by bronchoscopy and cultural proof of the pathogen followed by an adequate antifungal treatment. PMID- 12588485 TI - Primary cutaneous aspergillosis in an immunocompetent patient. AB - A case of primary cutaneous aspergillosis in a 39-year-old woman with apparently normal immune status is reported. The woman had an occasionally suppurating nodular lesion which developed in the site of a trauma on the back of the right wrist. Diagnosis was based on histological and mycological examination. Systemic involvement was not found. Surgical excision led to clinical and mycological recovery, confirmed at follow-up 18 months later. PMID- 12588486 TI - Tinea barbae caused by a zoophilic strain of Trichophyton interdigitale. AB - A deep absceding infection is reported of the inframandibular part of the face of a 22-year-old male student due to a zoophilic strain of Trichophyton interdigitale. The fungus was probably acquired from the cat of the patient. Initial therapy by a general practitioner was with topical glucocorticoids and oral antihistaminica. The patient developed a severe phlegmoneous inflammation of the bearded part of the face. Later, the patient was successfully treated by a combination of itraconazole and fluconazole. Identification of the species was confirmed by light and scanning microscopy as well as sequence analysis of the internal transcribed spacer region of the ribosomal DNA. PMID- 12588487 TI - An unusual case of tinea capitis due to Trichophyton soudanense. AB - A case of tinea capitis due to Trichophyton soudanense observed in a 4-year-old African girl is described. The infection manifested with diffuse pustular lesions of the scalp. Diagnosis was based on culture. Genome study by single primer PCR fingerprinting was also performed. A younger sister, aged 10 months, was the healthy carrier of the mycete. PMID- 12588488 TI - Mycosis on mycosis fungoides: zoophilic dermatophytosis selectively superimposed on pre-existing cutaneous T-cell lymphoma (mycosis fungoides) plaques. AB - We report a case of tinea corporis caused by a cattle-derived strain of Trichophyton mentagrophytes in a 44-year-old male affected by cutaneous T-cell lymphoma (CTCL, so-called mycosis fungoides). Fungal colonization of glabrous skin was strictly confined within pre-existing lymphomatous plaques. Either oral itraconazole or griseofulvin, or topical terbinafine were ineffective until the patient, who was treated with systemic retinoids and interferon-alpha for his CTCL, was shifted from leucocyte to lymphoblastoid interferon. The hypothesis that a local immunodisturbance could be responsible for the selective superimposition of tinea on CTCL lesions ('mycosis on mycosis'), and that such an immunodisturbance could be partially corrected by the interferon switch is discussed. PMID- 12588489 TI - Fatal peritonitis due to Trichoderma sp. in a patient undergoing continuous ambulatory peritoneal dialysis. AB - We describe a rare case of peritonitis caused by an unusual fungus, Trichoderma sp., in a patient on continuous ambulatory peritoneal dialysis. Management of the patient consisted of Tenckhoff catheter removal and antifungal chemotherapy, but the patient died. PMID- 12588492 TI - Measurement of condylar motion: a plea for the use of the condylar kinematic centre. AB - In this study, a plea is given for the use of the kinematic centre in studies of the kinematics of the human temporomandibular condyle. The concept of the kinematic centre is based upon the assumption that the movements of the condyle disc complex within the temporomandibular joint can reasonably well be described by those of a ball-shaped condyle-disc complex. The kinematic centre is then the centre of the sphere. Its movement traces have the advantage that they are smooth and have a good reproducibility between consecutive movements. Moreover, the open and close traces are just a few tenths of a millimetre apart and show no crossings. This makes the kinematic centre a suitable choice in order to avoid false-positive diagnoses in the study of internal derangements by means of condylar movement recordings. However, the kinematic centre has the disadvantage that the mandibular movements have to be recorded by rather complicated six degrees of freedom recording equipment and that the exploration algorithm for its location may sometimes have difficulties in finding the right location. PMID- 12588493 TI - The effect of the addition of poly(methyl methacrylate) fibres on some properties of high strength heat-cured acrylic resin denture base material. AB - The self-reinforcement of acrylic resin with butadiene styrene surface treated poly(methyl methacrylate) fibres has been reported to have the potential to substantially improve the transverse bend strength of conventional heat-cured acrylic resin. The aim of this study was to investigate the effect of the addition of butadiene styrene surface treated poly(methyl methacrylate) fibres in cross-ply arrangement to high impact acrylic resin on the transverse and impact strength. Specimens were prepared as specified in the International Standard Organization and British Standards for the Testing of Denture Base Resins (ISO 1567, 1988; BS 2487, 1989) and the British Standard Specification for Orthodontic resins (BS 6747, 1987) for transverse bend and impact testing. The impact strength was measured using a Zwick pendulum impact tester and the transverse bend strength measured using a Lloyds Instruments testing machine. The results showed that the impact strength was not improved with the addition of fibres, high impact acrylic resin with fibres (LF) 11.1 kJ m-2 and high impact acrylic resin (L) (12.5 kJ m-2). The modulus of rupture was decreased with the addition of fibres (57.8 MPa) for (LF) compared with (60.4 MPa) for (L). The modulus of elasticity was also reduced with the addition of fibres (1834.9 MPa) (LF) and 2086.2 MPa (L) as was the peak load (LF) (50.8 N) and (L) (55.8 N). It was concluded that the addition of surface treated poly(methyl methacrylate) fibres in cross-ply arrangement to high strength acrylic resin did not produce an improvement in the impact or transverse strength and cannot be recommended as a method of reinforcement. PMID- 12588494 TI - Marginal gap formation of composites in dentine: effect of water storage. AB - This study investigated the effects of water storage on the marginal adaptation of two composite resins (Spectrum TPH and Ariston AT), two polyacid-modified composite resins (Dyract AP and F2000) and a new PRG composite (Reactmer) to dentine over time. Two cylindrical dentine cavities (1.5 mm diameter and 1.5 mm deep) were prepared on the horizontally sectioned surfaces of freshly extracted teeth. In each tooth, one cavity was restored using composites with their respective bonding system and the other without the bonding system. The sample size for each material with/without bonding system was 6. The composites were placed in one increment, bulk-polymerized, immediately finished/polished with the Sof-lex discs system and fine polished on 30 microm silicon carbide/9 microm aluminium oxide lapping film discs in a microgrinding system. The specimens were then stored in distilled water at 37 degrees C and the maximum marginal gap width between the material and the dentine wall was determined at 24 h, 1 week, 2 weeks, 3 weeks and 4 weeks using a measurescope at x500 magnification. Results were subjected to statistical analysis using monva, anova/Scheffe's post hoc test and independent samples of t-test at significance level 0.05. At all time intervals, no significant difference in marginal gap formation was observed between materials. Where bonding systems were not used, a decrease in gap widths over time was observed with most materials but only Dyract AP exhibited a significant decrease. The latter was observed after 2 weeks storage in water. The use of bonding systems reduced dentine marginal gaps significantly and is therefore mandatory for all composites evaluated. Marginal gaps arising from polymerization contraction of conventional, polyacid-modified and PRG composites cannot be fully compensated by hydroscopic expansion. Clinical relevance Bonding systems are mandatory for all composites when bonding to dentin. In the event of a bond failure at placement, hygroscopic expansion of composites can reduce marginal gaps but not completely. PMID- 12588495 TI - Biofilm formation of Candida albicans on the surfaces of deteriorated soft denture lining materials caused by denture cleansers in vitro. AB - Candidal colonization and subsequent biofilm formation on denture materials are important in the development of pathogenesis, such as denture stomatitis. Routine use of denture cleansers is one of the most effective methods of denture plaque control, although the incompatibility of soft liners and denture cleansers cause damage to the materials. The present study, biofilm formation of Candida albicans on the surfaces of soft denture lining materials, immersed in denture cleansers for 180 days were studied. Seven commercially available soft denture lining materials, were artificially deteriorated by immersion into three commercially available denture cleansers for 180 days, and subsequent fungal growth and biofilm formation were studied by measuring pH of the media and by the use of adenosine triphosphate (ATP) analysis. Fungal biofilm formation on the deteriorated soft liners varied depending upon the combination of the soft liners and denture cleansers. Several combinations of soft liners with denture cleansers exhibited the significantly high colonization capacity as compared with each sample immersed in distilled water, used as individual controls. The relationship between the biofilm formation on the samples of each material and the surface roughness of the soft lining materials was analyzed. However, no significant correlation was observed. The results, taken together, suggested that fungal colonization could be predominantly regulated by the combination of lining material with denture cleansers. In clinical terms, our findings suggests that daily cleansing of soft lining materials with mismatched denture cleansers promoted the subsequent biofilm formation of fungi on the materials. PMID- 12588496 TI - Experimental salivary pellicles formed on the surface of self-curing resin. AB - The aim of this study was to identify the salivary components present in the pellicles formed on self-curing resin and to investigate the qualitative variations in adsorbed salivary pellicle compositions according to different exposure time to saliva. Experimental pellicles were formed by the incubation of polymerized resin particles with fresh human parotid or submandibular-sublingual saliva for either 20 min or 2 h. Pellicles were extracted using formic acid and lyophilized, they were then subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblotting to identify the adsorbed salivary components. The amino acid profiles of the 2 h-pellicles were analysed and compared with those of fresh glandular salivas. There was a difference in the 2 h pellicle components on the self-curing resin compared with those of other dental materials as well as tooth enamel. The amino acid profiles of the 2 h-pellicles were also different from those of fresh glandular salivas. In the case of submandibular-sublingual saliva, the components of the 2 h-pellicle showed a different pattern compared with those of the 20 min-pellicle. However, there was no significant difference between the components of the 2 h- and 20 min-pellicles in the case of parotid saliva. A distinct difference was found in the surface binding affinities of immunoglobulin (IgA) from different glandular salivas. The findings of this study provide information concerning the initial bacterial adhesion on the surfaces of self-curing resin. PMID- 12588497 TI - Correlation between periodontal status and biting ability in Chinese adult population. AB - This study investigated the relation between periodontal condition and biting ability in a Chinese population using the pressure-detecting sheet. A total of 142 subjects residing in Nanchang, Jiangxi province, China, participated in the study. The examination included probing pocket depth (PD), clinical attachment level (CAL), bleeding on probing and coronal caries teeth. Biting abilities per person, biting force, biting pressure and occlusal contact area were measured using the sheet. Number of teeth present showed positive correlation with biting force and occlusal contact area, but were negatively correlated with biting pressure. No significant correlation was observed between the mean PD, percentage of pocket > or = 4 mm, bleeding index and any of biting abilities. The mean CAL showed a significantly negative correlation with biting force and occlusal contact area. Multiple stepwise regression analysis selected the number of teeth present, sex and age as the significant factors affecting the biting ability. However, no periodontal indices were picked up as the significantly contributing factor. There appears to be little effect of periodontal condition on biting ability. Ageing was selected as a contributing factor to reduction in the Chinese adults, but not in the Japanese population. PMID- 12588498 TI - Fracture reasons in ceramic-fused-to-metal restorations. AB - Ceramic-fused-to-metal restorations are widely used in dentistry with a high degree of general success. Fracture of the ceramic veneers as a result of oral function or trauma is not an uncommon problem in clinical practice. Although fractures of such restorations do not necessarily mean the failure of the restoration, the renewal process is both costly and time consuming and therefore remains a clinical problem. Fractures in the anterior region pose an aesthetic problem but when they are in the posterior, chewing function could also be affected. The published literature reveals that reasons for failures cover a wide spectrum from iatrogenic factors to laboratory mistakes or because of factors related to the inherent structure of the ceramics or simply to trauma. PMID- 12588499 TI - Time-series analyses of mandibular and perioral soft tissue movements during mastication. AB - Masticatory movements are rhythmically repeated and coordinated movements of the jaw, tongue and facial muscles. Thus, we considered that the elucidation of movements that are specific to perioral soft tissue, as a result of perioral facial muscle activities, should be useful for evaluation of the smoothness of masticatory movements. The aim of this study was to evaluate the smoothness of masticatory movements from the component of movements that are specific to perioral soft tissue during mastication by the application of time-series analysis. The subjects were 15 healthy persons with complete natural dentition. The experimental food used for mastication in this study was sufficiently softened chewing gum. The results showed that the component of movements that are specific to perioral soft tissue during mastication are the equal repetition spatially and stable movements temporally, and that these movements have the same accurate rhythm as that of mandibular movements and cooperate with mandibular movements temporally. Moreover, the results suggested, from the viewpoint of kinematics, that the innervation of the central pattern generator was concerned with the neural basis of rhythm generation of perioral facial muscles. Therefore, the component of movements that are specific to perioral soft tissue during mastication is useful for evaluation of the smoothness of masticatory movements. PMID- 12588500 TI - Association between masticatory performance and maximal occlusal force in young men. AB - The purpose of this study was to examine the relationship between masticatory performance and maximal occlusal force in dentate subjects, using test foods of varying hardness. Subjects were 20 young males with natural dentitions. Gummy jellies with two different degrees of hardness have been used as test foods to evaluate masticatory performance. Masticatory performance was evaluated by the increase of the surface area of expectorated pieces of comminuted gummy jelly that was calculated from the concentration of gelatin. Maximal occlusal force was measured with pressure sensitive sheets (DENTAL PRESCALE 50H R type). The results suggested that maximal occlusal force had a significant positive correlation to the masticatory performance both with soft gummy jelly and hard gummy jelly. The correlation coefficient (r) with hand gummy jelly tended to be larger than with soft gummy jelly. PMID- 12588501 TI - Prevalence study of signs and symptoms of temporomandibular disorders in university students. AB - The aim of this study was to evaluate the prevalence of signs and symptoms of temporomandibular disorders (TMD) in university students from 19 to 25 years old, male and female, through the distribution of frequency of the data obtained from a questionnaire and physical examination. The results showed that 68% of the subjects exhibited some degree of TMD, and the women were the most affected. Signs and symptoms such as articular sounds, pain to palpation of the masticatory, cervical and scapular girdle musculature, subjective sensation of emotional stress, and postural and occlusal changes were more evident in the group with TMD, although they were also present in subjects classified as TMD free. Limitations in the mandibular movements were not found. The association of the obtained data allowed identifying a high prevalence of signs and symptoms of TMD in the Brazilian university population. PMID- 12588502 TI - The effect of disinfectant agents in eliminating the contamination of dental unit water. AB - High concentrations of water-borne organisms cause multiple public health problems. Contamination of water exiting the dental unit water lines could be inhibited with the use of some disinfectants. The purpose of this investigation was to establish the effect of two disinfectants and to test their capacity to eliminate colony forming units (CFU) per mL. Vacuum lines of four chairs were treated for a total of 2 weeks. Two disinfectants (Bio 2000 and Alpron) were used as per manufacturer's instructions. Water samples for hetereothrophic counts from each unit's air/water syringe line were collected before treating the first patient of the day. Baseline, daily, first and second week samples of 10 mL were plated on blood agar plates and eosin ethylene blue agar. For meosifilic bacterial counts, Mueller Hinton agar plates with 1 mL direct and 1/10 were used in sterile serum and CFU were counted. The suspected colonies were further evaluated using API 20E and API 20NE. No Gram(-) opportunistic pathogens were found during the entire observation. Baseline contamination level (>102 CFU mL-1) without use of disinfectants was significantly higher (P < 0.0001) than at both first and second weeks when disinfectants were used. No colony was formed when Bio 2000 was used after both first and second weeks, whereas small number of CFU mL-1 was found at the end of the first week when Alpron (<10) was used. In conclusion, when used daily, both disinfectants prevent the development of bacterial contamination after first and second weeks with no significant differences (P=0.35). PMID- 12588503 TI - The relationship between the occlusal interference side and the symptomatic side in temporomandibular disorders. AB - This study aimed to investigate the relationship between the facial side of pain or clicking symptoms and the side of occlusal interference, and to examine the features of pain and clicking patients in terms of frequencies of occlusal interferences in relation to the symptomatic side. Occlusal conditions in 31 pain patients and 40 clicking patients were investigated just after the relief of pain or clicking solely by means of bite plane therapy. The symptomatic side did not associate with the side of fewer occlusal contacts in intercuspal position, no canine contact on the working side, occlusal contact on the non-working side, or unilateral contact in retruded contact position. The results of this study suggest that pain or clicking does not necessarily tend to appear on the side of these occlusal interferences. Additionally, characteristic inclination for pain and clicking patients in terms of frequencies of occlusal interferences in relation to the symptomatic side was not identified. PMID- 12588504 TI - Influence of working side contacts on masticatory function for mandibular distal extension removable partial dentures. AB - The aim of this study was to clarify the influence of working side contacts for masticatory function for a distal extension removable partial denture. Five subjects who had edentulous arches from second premolar to second molar opposing natural teeth participated. Working side contacts were altered by using three types of artificial teeth attached to the denture base. A-form teeth made contact with opposite teeth while acting as the working side in a lateral excursion. The cusp angles of B- and C-form teeth were decreased by 10 degrees and 20 degrees, respectively compared with A-form teeth. B-and C-form teeth discluded with opposite teeth on the working side. Subjects were asked to perform two kinds of masticatory performance test. Obtained data were evaluated by the repeated measures anova and the Tukey test for multiple comparisons. Results revealed that there was a significant difference (P=0.001) in the three types of teeth in the mixing ability test and the value of A-form teeth was statistically less value than those of the other teeth. However, there was no significant difference in the comminuting ability test. These results suggested that working side contacts between artificial teeth and opposite natural teeth influenced mixing ability, but not comminuting ability. PMID- 12588505 TI - Relation between clenching strength and occlusal force distribution in primary dentition. AB - The aim of this study was to investigate the relation between clenching strength and occlusal force distribution in primary dentition. Twenty healthy children with normal occlusions: 11 boys and 9 girls, ages 3.2-5.8 years (avg. 4.5 years) were selected. Setting the bilateral masseter muscular activity at maximum clenching in full intercuspation as 100%, the occluding forces at 20, 40, 60, 80 and 100% clenching were recorded with pressure-sensitive sheets (Dental Prescale 50H, type R, Fuji Photo Film Co.), and the force of each primary tooth was analysed by computer (Occluzer FPD703). Occlusal force distribution was expressed as a percentage of the total occlusal force of each tooth and was compared between each clenching. There were no significant differences between various clenching strengths in the occlusal force distribution in primary dentition [one way repeated-measures analysis of variance (ANOVA)]. Thus, the results of the present study suggest that the distribution of occluding forces on a primary dental arch had its own pattern and that the clenching strength had no effect on that pattern. These patterns may be useful in determining occlusal function in children. PMID- 12588506 TI - The effect of custom tray material type and fabrication technique on tensile bond strength of impression material adhesive systems. AB - The purpose of this investigation was to evaluate the tensile bond strengths of two impression material systems (polysulphide and polyvinyl siloxane) to two custom tray materials [autopolymerizing and visible light-cured (VLC) acrylic resin]. The effect of polymerizing the tray materials directly against wax spacer and tin foil was evaluated for each material. Polymerizing tray materials against tin foil significantly increased the bond strengths of polysulphide and polyvinyl siloxane impression materials to VLC and autopolymerizing acrylic resin tray materials. Polyvinyl siloxane VLC cured against tin foil combination produced the strongest bond. The VLC resin tray material generated greater bond strength than autopolymerizing acrylic resin when polymerized against tin foil. PMID- 12588507 TI - Tooth contacts at the mandibular retruded position, influence of operator's skill on bite registration. AB - Occlusal conditions at the retruded contact position (RCP) have been thought to be among the aetiological factors for temporomandibular disorders. However, the role of these occlusal factors still remain unclear. The purpose of a series of studies was to investigate the locations of RCP contacts and to discuss their implication by the authors. In the study reported here, the influence of the operator's skill on bite registration was surveyed. Eighty-six dental students were selected as subjects. One specialist in prosthetic dentistry moved the subject's mandible up and down while guiding it gently backward until it rotated on its retruded axis. Once the initial tooth contacts were established, the operator checked them by using a thin articulating paper (direct method). On the other hand, six younger operators following precise instruction from the specialist about the technique of jaw guidance examined each of the subjects for location of RCP contacts using the indirect method. Silicone-based impression material was used for bite registration while the subject kept the jaw at this contact position (indirect method). Wilcoxon signed rank test was applied to evaluate differences between the expert operator and six younger operators regarding locations of RCP contacts. Results from the expert operator (direct method) showed that tooth contact at the first premolar was most frequently observed and the number of subjects who possessed unilateral tooth contacts was more than that with bilateral tooth contacts. On the other hand, in the case of the indirect method performed by younger operators, tooth contact patterns significantly differed from those of the direct method. PMID- 12588508 TI - Effect of nitrous oxide on dental patients with cerebral palsy -- using an electromyogram (EMG) from orofacial muscles as an index. AB - The efficacy of nitrous oxide (N2O) inhalation on the elevated orofacial muscle tonus during dental treatment in patients with cerebral palsy (CP) was studied by measuring the mean frequency of electromyographic (EMG) discharge, blood pressure (BP) and heart rate (HR) taking mentally retarded patients (MR) as a control. At a pre-treatment stage, the mean frequency of the EMG was higher in the CP patients than in the MR patients. By forcing the jaw open, the mean frequency of the EMG, BP and HR was elevated significantly in both types of patients. The enhanced level of the mean frequency of the EMG was higher in the CP patients than in the MR patients. After inhalation of N2O, the mean frequency was reduced significantly in both types of patients, however, the reduction rate was higher in the CP patients than the MR patients; BP and HR were not affected. These results demonstrate the selective efficacy of N2O in reducing the orofacial muscle tonus in CP patients during dental treatment probably because of the N2O inhibiting the function of the central nervous system. The present study therefore suggests the beneficial properties of N2O induced sedation performed during dental treatment on CP patients. PMID- 12588509 TI - Cocaine- and amphetamine-regulated transcript is present in hypothalamic neuroendocrine neurones and is released to the hypothalamic-pituitary portal circuit. AB - Cocaine- and amphetamine-regulated transcript (CART) is present in a number of hypothalamic nuclei. Besides actions in circuits regulating feeding behaviour and stress responses, the hypothalamic functions of CART are largely unknown. We report that CART immunoreactivity is present in hypothalamic neuroendocrine neurones. Adult male rats received a systemic injection of the neuronal tracer Fluorogold (FG) 2 days before fixation, and subsequent double- and triple labelling immunoflourescence analysis demonstrated that neuroendocrine CART containing neurones were present in the anteroventral periventricular, supraoptic, paraventricular (PVN) and periventricular nuclei of the hypothalamus. In the PVN, CART-positive neuroendocrine neurones were found in all of cytoarchitectonically identified nuclei. In the periventricular nucleus, approximately one-third of somatostatin cells were also CART-immunoreactive. In the medial parvicellular subnucleus of the PVN, CART and FG coexisted with thyrotrophin-releasing hormone, whereas very few of the corticotrophin-releasing hormone containing cells were CART-immunoreactive. In the arcuate nucleus, CART was extensively colocalized with pro-opiomelanocortin in the ventrolateral part, but completely absent from neuroendocrine neurones of the dorsomedial part. To assess the possible role of CART as a hypothalamic-releasing factor, immunoreactive CART was measured in blood samples from the long portal vessels connecting the median eminence with the anterior pituitary gland. Adult male rats were anaesthetized and the infundibular stalk exposed via a transpharyngeal approach. The long portal vessels were transected and blood collected in 30-min periods (one prestimulatory and three poststimulatory periods). Compared to systemic venous plasma samples, baseline concentrations of immunoreactive CART were elevated in portal plasma. Exposure to sodium nitroprusside hypotension triggered a two-fold elevation of portal CART42-89 immunoreactivity throughout the 90-min stimulation period. In contrast, the concentration of portal plasma CART immunoreactivity dropped in the vehicle infused rats. The current study provides further evidence that CART is a neuroendocrine-releasing factor with a possible impact on anterior pituitary function during states of haemodynamic stress. PMID- 12588510 TI - Opioid receptor subtypes involved in the regulation of prolactin secretion during pregnancy and lactation. AB - Afferent endogenous opioid neuronal systems facilitate prolactin secretion in a number of physiological conditions including pregnancy and lactation, by decreasing tuberoinfundibular dopamine (TIDA) inhibitory tone. The aim of this study was to investigate the opioid receptor subtypes involved in regulating TIDA neuronal activity and therefore facilitating prolactin secretion during early pregnancy, late pregnancy and lactation in rats. Selective opioid receptor antagonists nor-binaltorphimine (kappa-receptor antagonist, 15 micro g/5 micro l), beta funaltrexamine (mu-receptor antagonist, 5 microg/5 microl) and naltrindole (delta-receptor antagonist, 5 microg/5 microl) or saline were administered intracerebroventricularly (i.c.v.) on day 8 of pregnancy during a nocturnal prolactin surge, on day 21 of pregnancy during the ante partum prolactin surge or on day 7 of lactation before the onset of a suckling stimulus. Serial blood samples were collected at regular time intervals, via chronic indwelling jugular cannulae, before and after drug administration and plasma prolactin was determined by radioimmunoassay. TIDA neuronal activity was measured using the 3,4-dihydroxyphenylacetic acid (DOPAC) : dopamine ratio in the median eminence 2 h 30 min after i.c.v. drug injection. In each experimental condition, plasma prolactin was significantly inhibited by both kappa- and mu-receptor antagonists, whereas the delta-receptor antagonist had no effect compared to saline-injected controls. Similarly, nor-binaltorphimine and beta funaltrexamine significantly increased the median eminence DOPAC : dopamine ratio during early and late pregnancy, and lactation whereas naltrindole had no effect compared to saline-injected controls. These data suggest that TIDA neuronal activity, and subsequent prolactin secretion, is regulated by endogenous opioid peptides acting at both kappa- and mu-opioid receptors during prolactin surges of early pregnancy, late pregnancy and lactation. PMID- 12588511 TI - Involvement of the entorhinal cortex in the stress response to immobilization, but not to insulin-induced hypoglycaemia. AB - Although the involvement of the limbic system in the neuroendocrine responses to some stressors has been documented, the specific role of the entorhinal cortex has not been elucidated. In this study, we investigated the involvement of the entorhinal cortex in stress responses. Fos immunoreactivity, a widely used marker for neuronal activation, was detected in the entorhinal cortex of rats subjected to immobilization stress, whereas no marked staining was observed in the entorhinal cortex of the control and insulin-induced hypoglycaemia groups. Lesion of the entorhinal cortex produced by ibotenic acid significantly attenuated the adrenocorticotropic hormone (ACTH) release evoked by immobilization; however, no significant change in ACTH release was observed in insulin-induced hypoglycaemia. No significant difference between entorhinal-lesioned rats and control rats was observed in blood glucose concentrations when subjected to either immobilization or to insulin-induced hypoglycaemia. Together, these results indicate that the entorhinal cortex is closely involved in the stress response to immobilization but not to insulin-induced hypoglycaemia. PMID- 12588512 TI - Serotonin receptors involved in vasopressin and oxytocin secretion. AB - Serotonin (5-HT), 5-HT agonists, the 5-HT precursor 5-hydroxytryptophan, 5-HT releasers and -reuptake inhibitors stimulate the release of vasopressin and oxytocin. We investigated the involvement of 5-HT receptors in the serotonergic regulation of vasopressin and oxytocin secretion. Vasopressin and oxytocin secretion was stimulated by 5-HT, the 5-HT(1A+1B+5A+7) agonist 5 carboxamidotryptamine (5-CT), the 5-HT(2A+2C) agonist DOI, the 5-HT(2C+2A) agonist mCPP, the 5-HT(2C) agonist MK-212, the 5-HT(3) agonist SR 57277 and the 5 HT(4) agonist RS 67506. The 5-HT(1A) agonist 8-OH-DPAT, which had no effect on vasopressin secretion, stimulated oxytocin secretion. The 5-HT-induced release of vasopressin and oxytocin was inhibited by central infusion of the 5-HT antagonists WAY 100635 (5-HT(1A)), LY 53857 (5-HT(2A+2C)), ICS 205-930 (5 HT(3+4)) and RS 23597 (5-HT(4)). The 5-HT2+6+7 antagonist metergoline in combination with the 5-HT1A+2+7 antagonist methysergide inhibited the stimulatory effect of 5-CT on both hormones, whereas the 5-HT1A+1B antagonist cyanopindolol only inhibited the oxytocin response. The 5-HT(2A) antagonist 4-(4-flourobenzoyl) 1-(4-phenylbutyl)-piperidine oxalate had no effect on DOI-induced hormone response. The 5-HT(2C) antagonist Y 25130 partly inhibited the stimulating effect of MK-212. ICS 205-930 and RS 23597 inhibited vasopressin and oxytocin secretion induced by RS 67506. WAY 100635 inhibited 8-OH-DPAT-induced oxytocin secretion. We conclude that 5-HT-induced vasopressin secretion primarily is mediated via 5 HT(2C), 5-HT(4) and 5-HT(7) receptors, whereas 5-HT(2A), 5-HT(3) and 5-HT(5A) receptors seem to be of minor importance. 5-HT-induced oxytocin secretion involves 5-HT(1A), 5-HT(2C) and 5-HT(4) receptors; in addition an involvement of 5-HT(1B), 5-HT(5A) and 5-HT(7) receptors seems likely, whereas 5-HT(2A) and 5 HT(3) receptors seem to be less important. PMID- 12588513 TI - Inducible nitric oxide synthase is responsible for nitric oxide release from murine pituicytes. AB - This study investigated whether pituicytes were able to produce and release nitric oxide (NO), and which type of nitric oxide synthase (NOS) would be responsible for this phenomenon. Lipopolysaccharide (LPS) 1 micro g/ml was used as inflammatory mediator. Because pituicytes are known to secrete interleukin (IL)-6 upon stimulation with LPS, this parameter was also investigated. Cultured pituicytes, from 4-week-old male mice, were stimulated with LPS for 6 h or 24 h. At 24 h, there was a significant increase in accumulated nitrite indicating NO formation. In contrast, IL-6 release was already significantly higher 6 h after stimulation and further increased at 24 h. The correlation between accumulated nitrite and secreted IL-6 was 0.84 after 24 h of incubation with LPS. The expression of inducible NOS (iNOS) mRNA in the pituicytes was significantly higher than the control level after 6 h and 24 h of exposure to LPS, with levels at 6 h being significantly higher than those at 24 h. There was no detected expression of endothelial NOS or neuronal NOS mRNA. Cultured pituicytes were also subjected to immunocytochemistry for iNOS protein at 6, 12, and 24 h after stimulation with LPS. Most cells were positive for iNOS, but there were no observable differences with the time points that we used. Collectively, these results show that pituicytes are able to produce NO, and that the inducible form of NOS is responsible for this production. Furthermore, there is a weak correlation between NO and IL-6 released from pituicytes after 24 h of stimulation with LPS. PMID- 12588514 TI - Genetic selection for coping style predicts stressor susceptibility. AB - Genetically selected aggressive (SAL) and nonaggressive (LAL) male wild house mice which show distinctly different coping styles, also display a differential regulation of the hypothalamic-pituitary-adrenal axis after exposure to an acute stressor. To test the hypothesis that coping style predicts stressor susceptibility, the present study examined line differences in response to a chronic stressor. Chronic psychosocial stress was evoked using two paradigms. In the first paradigm, a SAL or LAL male was living in sensory contact (except tactile contact) with a dominant SAL male for 25 days (sensory contact stress). In the second paradigm, a SAL or LAL male was, in addition to the first paradigm, defeated by a SAL male for 21 consecutive days (defeat stress). The sensory contact stressor induced in LAL mice chronic body weight loss and increased plasma adrenocorticotropic hormone levels compared to SAL mice and increased corticosterone levels, thymus involution and lower hippocampal mineralocorticoid receptor (MR) : glucocorticoid receptor (GR) ratio compared to LAL controls. The defeat stressor increased corticosterone secretion and caused adrenal hypertrophy and thymus involution in both mouse lines. Defeated LAL mice showed long-lasting body weight loss and higher corticosterone concentrations than SAL mice and lower hippocampal MR : GR ratio and decreased immobility behaviour in the forced swimming test than LAL controls. Hypothalamic corticotropin-releasing hormone mRNA expression was higher in defeated SAL than in controls. The present data show that both stress paradigms induced line-dependent physiological and neuroendocrine changes, but that the sensory contact stressor produced chronic stress symptoms in LAL mice only. This latter stress paradigm therefore seems promising to analyse the role of genetic factors in the individual differences in stress-related psychopathology. PMID- 12588515 TI - Effect of direct injection of melanin-concentrating hormone into the paraventricular nucleus: further evidence for a stimulatory role in the adrenal axis via SLC-1. AB - Melanin-concentrating hormone (MCH) is implicated in the control of a number of hormonal axes including the hypothalamic-pituitary adrenal (HPA) axis. Previous studies have shown that there is evidence for both a stimulatory and an inhibitory action on the HPA axis; therefore, we attempted to further characterize the effects of MCH on this axis. Intracerebroventricular injection of MCH increased circulating adrenocorticotropic hormone (ACTH) at 10 min post injection. Injection of MCH directly into the paraventricular nucleus (PVN) was found to increase both circulating ACTH and corticosterone 10 min after injection. Additionally, MCH was found to increase corticotropin-releasing factor (CRF) release from hypothalamic explants, and this effect was abolished by the specific SLC-1 antagonist SB-568849. Neuropeptide EI, a peptide from the same precursor as MCH was also found to increase CRF release from explants. These results suggest that MCH has a stimulatory role in the HPA axis via SLC-1, and that MCH exerts its effects predominantly through the PVN CRF neuronal populations PMID- 12588516 TI - In vitro effects of catecholamines and catecholestrogens on brain tyrosine hydroxylase activity and kinetics in the female catfish Heteropneustes fossilis. AB - Effects of catecholamines and catecholestrogens on tyrosine hydroxylase (TH) activity and kinetics were investigated in the telencephalon and hypothalamus of female Heteropneustes fossilis in gonad quiescent (resting) and recrudescent (preparatory) phases. Dopamine, noradrenaline and adrenaline and the catecholestrogen, 2-hydroxyestradiol-17 beta inhibited TH activity in a concentration-dependent manner in both resting and preparatory phases, with a higher effect in the resting phase. Two- methoxyestradiol-17 beta did not alter TH activity in any season. The catecholamines inhibited TH in a competitive manner increasing apparent K(m) values significantly without altering the apparent V(max). Two-hydroxyestradiol-17 beta inhibited significantly the enzyme in a noncompetitive manner and decreased apparent V(max) without altering apparent K(m) values. The apparent K(i) is higher for dopamine than noradrenaline or adrenaline. The apparent K(i) for 2-hydroxyestradiol-17 beta is not significantly different from that of noradrenaline. The present results suggest an interaction between oestradiol-17beta (E2) and catecholamine metabolism at the level of tyrosine hydroxylation and E2 effects on catecholamines may be mediated through its 2-hydroxylation. PMID- 12588517 TI - Growth hormone-releasing hormone-producing and dopaminergic neurones in the mouse arcuate nucleus are independently regulated populations. AB - Differentiation of hypophysiotropic neurones that regulate the secretion of growth hormone (GH) and prolactin is influenced by GH and prolactin. Genetic GH and prolactin deficiency in mutant rodent models such as the Ames dwarf (df/df) mouse results in an increase in the number of GH-stimulatory GH-releasing hormone (GHRH) neurones and a reduction of prolactin-inhibitory tuberoinfundibular dopaminergic (TIDA) neurones in the arcuate nucleus during postnatal development. The present study tested the hypothesis that these concomitant changes in numbers of tyrosine hydroxylase (TH)- and GHRH-immunoreactive neurones in df/df hypothalamus might represent a neuronal population of fixed number that undergoes a partial change in phenotype during postnatal development. To evaluate this possibility, the postnatal reduction of the df/df TIDA population was prevented by administering prolactin neonatally to preserve TH phenotype; dwarf and normal sibling mice were treated with daily injections of ovine prolactin or vehicle starting at postnatal day 12 and continuing for 30 days. Following this treatment, numbers of arcuate neurones containing GHRH or TH, or both, were quantified using immunocytochemistry. It was hypothesized that prolactin preservation of TH-immunoreactive cell number would be accompanied by either a decrease in the GHRH-producing population or an increase in numbers of cells producing both TH and GHRH. In prolactin-treated normal (DF/df) mice, numbers of arcuate TH-immunoreactive neurones were similar to those in vehicle-treated normals. Numbers of TH-positive neurones in prolactin-treated dwarfs were higher than in vehicle-treated dwarfs, and did not differ from numbers in DF/df. Numbers of GHRH-immunoreactive cells in vehicle-treated df/df were higher than in vehicle treated DF/df, and were not different in prolactin-treated groups of either dwarf or normal mice. Neurones containing both TH and GHRH constituted 15% of the TH population, and 76% of the GHRH population, in control normal mice; in control dwarfs, double-labelled cells were 9.3% of TH and 9.9% of GHRH. Numbers of cells immunoreactive for both TH and GHRH were not affected by prolactin treatment in either mouse type. These results demonstrate that the increase in number of GHRH expressing neurones in the df/df arcuate nucleus does not occur at the expense of the TH phenotype, and that this increase is not influenced by prolactin feedback. Although coexpression of TH and GHRH in a subpopulation indicates that TIDA and GHRH populations are not exclusive, they appear to be influenced independently by prolactin and GH signals during development. PMID- 12588518 TI - Gender specific effect of neonatal handling on stress reactivity of adolescent rats. AB - Early neonatal handling of rat pups produces dampened hypothalamic-pituitary adrenal axis reactivity to stress in adult male offspring. However, less is known about whether there is a similar effect for females. Although, most studies of neonatal handling have examined subsequent effects during adulthood, adolescence is an important developmental stage for stress responsivity. To address these issues, the effect of neonatal handling on the endocrine stress response and brain activity of male and female rats was determined in response to acute restraint stress during adolescence. Consistent with previous findings in adult males, neonatal handling reduced restraint stress-induced hormone levels in adolescent males. However, in contrast, we found elevated plasma hormone concentrations in handled females. A gender-specific handling effect on brain activity was also evident, with significantly increased stress-induced activation of the posterior cingulate cortex of handled females, as measured by c-fos mRNA expression. The striking gender difference in the effect of early neonatal handling provides evidence that this must be considered as an important variable in subsequent stress responsivity induced by early manipulations. PMID- 12588519 TI - Oestradiol-dependent and -independent modulation of tyrosine hydroxylase mRNA levels in subpopulations of A1 and A2 neurones with oestrogen receptor (ER)alpha and ER beta gene expression. AB - Oestradiol (E2) induces luteinizing hormone-releasing hormone (LHRH) hypersecretion, thereby triggering LH surge release in ovariectomized (OVX) rats. Neural signals responsible for the surge are marked by a morning increase in LHRH gene expression and an afternoon increase in LHRH release. Evidence suggests that subpopulations of noradrenergic neurones may be responsible for one or both of these signals. To further investigate this issue, we examined effects of E2 on the activity of A1 and A2 noradrenergic neurones, as reflected in changes in tyrosine hydroxylase (TH) mRNA expression, on the day of LH surge release. We then used dual-label in situ hybridization to determine whether E2-induced changes occurred primarily in A1 and A2 subdivisions wherein most noradrenergic neurones expressed oestrogen receptor (ER)alpha and/or ER beta mRNA. We found that in all subdivisions, levels of TH mRNA were higher in E2- than oil-treated rats at 12.00 h. These differences resulted from a decline in TH mRNA expression in oil-treated rats, as well as a rise in levels in E2-treated rats between 10.00 h and 12.00 h. During the afternoon, TH mRNA expression in most A1 and A2 subdivisions peaked at 14.00 h when LH surge release began. However, in all but the middle and caudal A2 subdivisons, levels were similar in E2-treated and control rats at this time. This was attributable to a widespread increase in TH mRNA expression between 12.00 h and 14.00 h in OVX rats. There was no evidence that E2 induced changes in TH mRNA expression preferentially in regions wherein most neurones contained ER alpha or ER beta mRNA. Our findings suggest that E2 activation of middle and caudal A2 neurones, in conjunction with the widespread E2-independent activation of noradrenergic neurones in other subdivisions, may play a role in the induction of LH surge release. PMID- 12588520 TI - NF kappa B activation in mouse pituitary: comparison of response to interleukin-1 beta and lipopolysaccharide. AB - The mouse anterior pituitary contains both types of interleukin (IL)-1 receptors, IL-1 receptor type I (IL-1RI) and IL-1 receptor type II (IL-1RII). These receptors are expressed mainly on somatotroph cells. In the present study, the ability of the mouse pituitary to respond in vivo to IL-1 or to lipopolysaccharide (LPS) was demonstrated by measuring, with an electrophoretic mobility shift assay, the presence of an active NF kappa B complex in cell nuclei from pituitaries of mice injected intraperitoneally with recombinant rat-IL-1 beta or LPS. Using immunohistochemistry with an antibody directed against the p65 NF kappa B subunit, a rapid and transient NF kappa B response to LPS was observed. This response was present predominantly in the nuclei of glial fibrillary acidic protein (GFAP)-positive cells and F4/80-labelled cells of the posterior and the anterior pituitary 15 min after stimulation and became faint after 2 h. In comparison, the early and strong NF kappa B response to IL-1 beta treatment was localized into somatotroph cells, GFAP positive cells and F4/80 labelled cells of the posterior and anterior pituitary. Activation of NF kappa B in response to IL-1 beta was no longer apparent in IL-1RI knockout mice, confirming that this receptor is essential for the transduction of IL-1 signal in the pituitary, but remained after LPS treatment. In addition, we investigated the effect of IL-1 on target genes by measuring the mRNA and proteins synthesis of growth hormone (GH), IL-6 and IL-1ra in the pituitary and the plasma. IL-1 beta was shown to induce a rapid and strong synthesis of IL-6 and IL-1ra in the pituitary but failed to regulate GH contents or release. These data suggest that the pituitary is able to respond to a systemic infection via cytokine-mediated responses transduced by IL-1. PMID- 12588521 TI - Daily rhythms in glucose metabolism: suprachiasmatic nucleus output to peripheral tissue. AB - The body has developed several control mechanisms to maintain plasma glucose concentrations within strict boundaries. Within those physiological boundaries, a clear daily rhythm in plasma glucose concentrations is present; this rhythm depends on the biological clock, which is located in the hypothalamic suprachiasmatic nucleus (SCN), and is independent of the daily rhythm in food intake. Interestingly, there is also a daily rhythm in glucose uptake, which also depends on the SCN and follows the same pattern as the daily rhythm in plasma glucose concentrations; both rise before the onset of activity. Thus, the SCN prepares the individual for the upcoming activity period in two different ways: by increasing plasma glucose concentrations and by facilitating tissue glucose uptake. In addition to this anticipation of glucose metabolism to expected glucose demands, the SCN also influences, depending on the time of the day, the responses of pancreas and liver to abrupt glucose changes (such as a glucose rise after a meal or hypoglycaemia). This review presents the view that the SCN uses different routes to (i) maintain daily glucose balance and (ii) set the level of the endocrine response to abrupt blood glucose changes. PMID- 12588522 TI - Puberty: mind and body. AB - Puberty is the attainment of fertility -- the ability to reproduce. It occurs because our brains begin to secrete one key hormone, gonadotropin-releasing hormone. This causes the pituitary gland to induce maturation of the testis and the ovary, which then produce sex steroid hormones and mature sperm and eggs. We should be concerned that the age at which puberty begins is decreasing in many countries. Puberty starts when adequate growth and energy storage has occurred, so the earlier age of puberty may reflect recent increases in adolescent obesity resulting from more sedentary lifestyles. PMID- 12588523 TI - A new system for laser-UVA-microirradiation of living cells. AB - Local nuclear irradiation of living cells has been used to gain insight into the dynamic changes that cell nuclei undergo in response to DNA damage. In particular, the effects of DNA double-strand breaks (DSBs), a major threat to the genomic integrity of cells, have been studied by local nuclear irradiation with ionizing radiation. This method has the disadvantage that it requires expensive equipment to generate a sufficiently high density of focused or collimated ionizing radiation. After appropriate sensitization of the cellular DNA, nuclear microirradiation with UVA can also produce DSBs. In this communication we present a semi-automatic system for laser-UVA-microirradiation based on a commercial laser scanning microscope. The system allows the convenient selection and precise irradiation of living cells, and could provide the basis for a more widespread availability of microirradiation facilities for DNA-repair research. PMID- 12588524 TI - Cryofixation of epithelial cells grown on sapphire coverslips by impact freezing. AB - Rapid cryofixation of cells cultured on coverslips without the use of chemical fixatives has proved advantageous for the immunolocalization of antigens by electron microscopy. Here, we demonstrate the application of sapphire-attached tissue culture cells (PtK2 epithelial cells and mouse myoblasts) to metal-mirror impact freezing. The potential of the Leica EM-CPC cryoworkstation for routine freezing and for safe transfer of the cryofrozen samples into a sapphire disc magazine for freeze-substitution (SD-FS unit) has been exploited. Subsequently, the SD-FS unit has been tested for its use in methanol freeze-substitution and low temperature embedding for immunoelectron microscopy. The structural preservation of Lowicryl HM20-embedded cells has been assessed as being free of damage by large ice crystals. PMID- 12588525 TI - The effect of the reinforcing carbon on the microstructure of pitch-based granular composites. AB - Carbon composites were prepared with four pitches (a commercial impregnating coal tar pitch, two thermally treated pitches and an air-blown pitch) and four granular carbons (anthracite, graphite, green petroleum coke and foundry coke). Granular carbon/pitch proportions were optimized for each composite and differed in the characteristics of the single components. Interactions of the pitch with the granular carbons during pyrolysis and their subsequent effects on the microstructure of the final composite were monitored by light microscopy. The results show that the light texture of the matrix and the porosity of the composite depend not only on the chemical composition of the pitch but also on the specific granular carbon used as reinforcing material. The same pitch may generate different light textures depending on the characteristics of the carbon. Composites from thermally treated pitches and graphite show highly ordered matrices orientated in the direction of graphite planes. Graphite particles seem to exert a huge influence on mesophase development during the pyrolysis of the treated pitches, affecting not only the orientation of the mesophase, but also reducing the rate of mesophase formation. On the other hand, when green petroleum coke is used with the thermally treated pitches, matrices show a small size light texture, due to the high reactivity of the pitch in the presence of this granular carbon. The porosity of the composites is controlled by both the pitch and the granular carbon. PMID- 12588526 TI - A novel light source for SICM-SNOM of living cells. AB - We have developed a novel light source for use in a scanning near-field optical microscope (SNOM or NSOM) based on a nanopipette whose distance from the sample surface is controlled using scanning ion conductance microscopy. The light source is based on the general principle of the chemical reaction between a fluorophore in the pipette and ligand in the bath, to produce a highly fluorescent complex that is continually renewed at the pipette tip. In these experiments we used fluo 3 and calcium, respectively. This complex is then excited with an Ar+ laser, focused on the pipette tip, to produce the light source. This method overcomes the transmission problem of more traditional SNOM probes and has been used to acquire simultaneous high-resolution topographic and optical images of biological samples in physiological buffer. A resolution of approximately 220 nm topographic and approximately 190 nm optical was determined through imaging fixed sea-urchin sperm flagella. Live A6 cells were also imaged, demonstrating the potential of this system for SNOM imaging of living cells. PMID- 12588527 TI - X-ray tomographic imaging of Ti/SiC composites. AB - In this paper, high-resolution tomographic synchrotron X-ray imaging is applied to study the occurrence and evolution of damage in Ti-6Al-4V/SCS6 SiC fibre composite materials. Three composite morphologies of increasing complexity have been studied, namely single fibre, single-ply and multi-ply composites. The single fibre composite was strained to full fibre fragmentation and the progressive introduction of damage monitored. For the single-ply composite, damage was introduced deliberately by laser drilling to establish the effect of damaged fibres on their neighbours, whereas for the multi-ply composite the morphology of a fibre bridging fatigue crack was studied. In addition to traditional mode I fibre fractures, subsequent fibre wedge cracks were observed presumably nucleating from damage introduced into the fibre surface by the first fracture event. In addition to these crack morphologies, spiral defects were observed for the single ply during failure. Finally, for the multi-ply composite, the matrix crack front showed a number of characteristic features, including advancement in fibre-free regions, crack bifurcation near fibres and different crack plane heights either side of a fibre. PMID- 12588528 TI - Characterization of the cutting edge of glass and diamond knives for ultramicrotomy by scanning force microscopy using cantilevers with a defined tip geometry. Part II. AB - The cutting edge of glass as well as diamond knives was studied at high resolution using a scanning force microscope (SFM). The local shape of the cutting edge was estimated from single line profiles of the SFM topographs taking into account the exact shape of the probing tip estimated by a high-resolution field emission scanning electron microscope (FESEM). The glass knives were prepared by 'balanced breaking'. The radius of the investigated cutting edges was found to be 3.2-4.4 nm and 4.3-6.0 nm for the 35 degrees and 45 degrees diamond knife, respectively, and 3.4-4.3 nm for the glass knives. Besides the opening angle and the cutting edge radius, the friction of a knife during sectioning represents a significant factor influencing the quality of sections. Thus, the roughness of both the diamond clearance angle side and the back side was characterized as well. Corresponding RMS values of the roughness were found to be smaller on the back side (approximately 0.14 nm) than on the clearance angle side (approximately 0.26 nm). PMID- 12588529 TI - Estimation of tissue layer level by sequential morphological reconstruction. AB - An automated method for labelling layers of clustered elements in two dimensional images based on morphological operations is presented here. Unlike the various distance transforms for regular lattices, the procedure deals successfully with clusters composed of different shaped and sized elements and provides a distance measure, in number of layers, from each element in the cluster to any other reference element (or elements). The method was applied to histological sections of two different types of polystratified epithelia. Local clusters in the different cell profile layers can subsequently be used to define the orientation of the tissue layers relative to each element in it. PMID- 12588530 TI - Precise 3D image alignment in micro-axial tomography. AB - Micro (micro-) axial tomography is a challenging technique in microscopy which improves quantitative imaging especially in cytogenetic applications by means of defined sample rotation under the microscope objective. The advantage of micro axial tomography is an effective improvement of the precision of distance measurements between point-like objects. Under certain circumstances, the effective (3D) resolution can be improved by optimized acquisition depending on subsequent, multi-perspective image recording of the same objects followed by reconstruction methods. This requires, however, a very precise alignment of the tilted views. We present a novel feature-based image alignment method with a precision better than the full width at half maximum of the point spread function. The features are the positions (centres of gravity) of all fluorescent objects observed in the images (e.g. cell nuclei, fluorescent signals inside cell nuclei, fluorescent beads, etc.). Thus, real alignment precision depends on the localization precision of these objects. The method automatically determines the corresponding objects in subsequently tilted perspectives using a weighted bipartite graph. The optimum transformation function is computed in a least squares manner based on the coordinates of the centres of gravity of the matched objects. The theoretically feasible precision of the method was calculated using computer-generated data and confirmed by tests on real image series obtained from data sets of 200 nm fluorescent nano-particles. The advantages of the proposed algorithm are its speed and accuracy, which means that if enough objects are included, the real alignment precision is better than the axial localization precision of a single object. The alignment precision can be assessed directly from the algorithm's output. Thus, the method can be applied not only for image alignment and object matching in tilted view series in order to reconstruct (3D) images, but also to validate the experimental performance (e.g. mechanical precision of the tilting). In practice, the key application of the method is an improvement of the effective spatial (3D) resolution, because the well-known spatial anisotropy in light microscopy can be overcome. This allows more precise distance measurements between point-like objects. PMID- 12588531 TI - A prism combination for near isotropic fluorescence excitation by total internal reflection. AB - Total internal reflection fluorescence (TIRF) microscopy is finding increasing application for selectively detecting molecules at or near a glass-water surface. As with all fluorescence methods, the efficiency of excitation of a fluorophore is potentially sensitive to the polarization state of the source. In TIRF, s polarized excitation produces an evanescent field that is perpendicular to the incident plane (y direction), whereas p-polarized light generates a more complex pattern but one dominated by a field that is vertical to the surface (z direction). Thus, fluorophores whose absorption dipoles are fixed in the x direction are not favourably aligned for excitation. Here we describe a beam splitting prism arrangement that allows excitation by two orthogonal beams, thus giving isotropic excitation in the x-y plane with s-polarized light. With linearly polarized light at the magic angle, near isotropic excitation in three dimensions should be achieved. This prism design should find application in polarized fluorescence microscopy to investigate the rotational motions of macromolecules or to minimize flickering of fluorescence emission arising from molecular rotations in single molecule studies. PMID- 12588533 TI - Aetiology, pathogenesis and treatment of sarcoidosis. PMID- 12588534 TI - Is there a role for microorganisms in the pathogenesis of sarcoidosis? AB - Sarcoidosis is a granulomatous disease that has the immunopathological features of being antigen-driven. It is a complex disease that appears to arise from the interaction of one or more triggers with an immunologically predisposed host. Previous reports of familial clustering and varying prevalence of sarcoidosis in different populations could reflect differences in ethnic predisposition or differences in local environmental exposures. This review focuses specifically on these areas that have been the subjects of intensive investigation recently. Specific focus is provided on the issue of an infective trigger and highlights popular candidates. It is concluded that microbes are a likely trigger (but not as an infection) in a genetically predisposed individual and that this initial event culminates in the sarcoidosis granulomatous response. PMID- 12588535 TI - The cytokine network in sarcoidosis and its clinical relevance. AB - In recent years, analysis of the cytokine network has substantially improved our knowledge of the immunopathogenesis of sarcoidosis. There is increasing evidence from clinical immunology that analysis of the cytokine network may be helpful for clinicians to assess the extent and activity of sarcoid inflammation. Genetic polymorphisms may contribute to interindividual differences in the regulation of cytokine release. Thus, disease phenotype-associated haplotypes should exist and their analysis might disclose risk profiles of individual patients. Furthermore, serological assessment of cytokines or soluble cytokine receptors may become suitable parameters in clinical practice to detect an ongoing inflammation in chronic sarcoidosis. PMID- 12588536 TI - Treatment of sarcoidosis -- from a basic science point of view. AB - Progress in our understanding of the scientific basis of granulomatous inflammation in sarcoidosis provides a framework for enlightened treatment decisions. Current evidence supports the concept that the pathogenesis of sarcoidosis involves a highly polarized T-helper 1 (Th1) immune response to pathogenic tissue antigens. Conventional treatment is focused on attenuating granuloma formation with antimalarial drugs that inhibit antigen presentation or with nonspecific anti-inflammatory agents such as glucocorticosteroids, methotrexate, or azathioprine. Anti-tumour necrosis factor (TNF)-alpha agents such as pentoxifylline, thalidomide, etanercept and remicade, have recently shown some successes in sarcoidosis. Designing future therapies depends on improved knowledge of the critical immunological processes operative in different stages of disease. PMID- 12588537 TI - Difficult treatment issues in sarcoidosis. AB - The management of sarcoidosis includes several crucial decisions. Not all patients with sarcoidosis need treatment. At least a third of patients will never be treated. It is unclear whether asymptomatic patients ever need therapy, even if they have extensive lung disease. One reason that clinicians are reluctant to start therapy is that many patients who are started on corticosteroids have a difficult time getting off therapy, even after 2 years. In the chronic patient, alternatives to corticosteroids have been developed. These include drugs such as methotrexate, azathioprine and hydroxychloroquine. These agents have been the standard second line of therapy for patients with chronic disease. However, these drugs do not always work. In addition, they are associated with their own toxicities. Another group of sarcoidosis patients have also emerged. These are the refractory patients, who have progressive disease whilst on therapy. For these patients, new agents such as thalidomide and the monoclonal antibodies to tumour necrosis factor have been occasionally helpful. This paper reviews several important issues in the management of sarcoidosis. PMID- 12588538 TI - Overview of the clinical efficacy of investigational anticancer drugs. AB - The purpose of this overview was to make a broad inventory of investigational drugs for medicinal cancer treatment and, specifically, to indicate the evidence of clinical efficacy. Information was retrieved from electronic database searches in Medline and CANCERLIT and relevant published reviews. As the most recent findings are first reported as conference abstracts, an important basis for identification of new drugs and clinical results was a hand search of 13,392 abstracts from five major recent cancer conferences. A total of 209 investigational approaches or drugs were identified and classified into one of eight groups according to proposed mechanism of action. For 28 drugs/approaches survival data were available from randomized controlled trials. Statistically significant benefit was observed for only 12. In earlier phases no or modest anticancer activity was reported. It is speculated that the expanding knowledge in tumour biology might not easily translate into new substantially better anticancer drugs. PMID- 12588539 TI - Duration of warfarin sodium therapy prior to electrical cardioversion of atrial fibrillation. AB - OBJECTIVES: To evaluate the duration of anticoagulation treatment with warfarin sodium before elective DC-cardioversion and to identify clinical variables predicting short-term versus long-term waiting times. DESIGN: Retrospective. SUBJECTS: Patients with a known start date for warfarin sodium, a known duration of atrial fibrillation (AF) and who underwent DC-cardioversion were included. MAIN OUTCOME MEASURES: Duration of treatment with warfarin sodium prior to DC cardioversion. METHODS: The hospital records of 288 consecutive patients with AF scheduled for elective cardioversion at two hospitals in Stockholm were reviewed. Only patients with a known start date for warfarin sodium and known duration of AF were included in the study. RESULTS: The median age was 70 (26-85) years and the duration of AF at time of cardioversion were 18 weeks (5-273) weeks. The median treatment duration prior to cardioversion with warfarin sodium was 12 weeks. Sinus rhythm was established in 224 (78%) patients of which 90 (40%) remained in sinus rhythm 1 month after cardioversion. In multivariate analysis, the only independent predictor of short waiting times for cardioversion (8 vs. 15 weeks) was if a cardiologist instituted the treatment with warfarin sodium (P < 0.001, 95% CI 5.0-9.0). CONCLUSION: The average waiting time from start of warfarin sodium treatment to elective cardioversion exceeds by far the recommended 3-4 weeks on therapeutic international normalized ratio (INR). In order to minimize the time period until cardioversion significant changes in the out-of-hospital care logistics has to be undertaken. PMID- 12588540 TI - Physical activity in relation to indices of endothelial function and angiogenesis factors in hypertension: a substudy of the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT). AB - BACKGROUND: Hypertension is an important risk factor for cardiovascular disease. The latest guidelines recommend regular physical exercise as initial step or adjunct in the treatment of hypertension. We investigated the association between physical activity and the degree of hypertension, as well as the relation to indices of endothelial damage/dysfunction and angiogenesis. METHODS: We studied 234 patients with hypertension (198 males; mean age 64 years; mean blood pressure 166/90 mmHg), who were compared with 60 age and sex-matched healthy normotensive controls. We assessed the patient's physical activity using the validated Baecke physical activity questionnaire and measured flow-mediated dilatation (FMD) of the brachial artery and von Willebrand factor (vWf) as indices of endothelial damage/dysfunction, whilst angiogenesis was assessed by measurement of plasma vascular endothelial growth factor (VEGF) and its soluble receptor (sFlt-1) both by ELISA. RESULTS: When hypertensive patients were compared with the controls, there was no statistically significant difference in total physical activity score using the Baecke questionnaire although plasma VEGF and vWf levels were higher, but sFlt-1 levels and FMD lower (all P < 0.001). Patients with high physical activity were younger, and had lower mean diastolic blood pressure and 10-year Framingham stroke risk, when compared with those with low physical activity; but indices of endothelial damage/dysfunction and angiogenesis were not significantly different. CONCLUSION: Physical activity scores in hypertensive patients are not significantly different from healthy normotensive controls, and there appears to be no relation to the abnormal processes of endothelial damage/dysfunction and angiogenesis seen in hypertensives. PMID- 12588541 TI - Atrial fibrillation or flutter and stroke: effectiveness of oral anticoagulation in clinical practice. PMID- 12588542 TI - Fermentative activity and production of volatile compounds by Saccharomyces grown in synthetic grape juice media deficient in assimilable nitrogen and/or pantothenic acid. AB - AIMS: To understand the impact of assimilable nitrogen and pantothenic acid on fermentation rate and synthesis of volatile compounds by Saccharomyces under fermentative conditions. METHODS AND RESULTS: A 2 x 3 factorial experimental design was employed with the concentrations of yeast assimilable nitrogen (YAN) (60 and 250 mg l(-1)) and pantothenic acid (10, 50 and 250 microg l(-1)) as variables. In media containing 250 microg l(-1) pantothenic acid, H2S production by two different species of Saccharomyces decreased when YAN was increased from 60 to 250 mg l(-1). Conversely, H2S production was significantly higher when the concentration of assimilable nitrogen was increased if pantothenic acid was deficient (10 or 50 microg l(-1)). Yeast synthesis of other volatile compounds were impacted by both assimilable nitrogen and pantothenic acid. CONCLUSIONS: While growth and fermentative rate of Saccharomyces was more influenced by nitrogen than by pantothenic acid, complicated interactions exist between these nutrients that affect the synthesis of volatile compounds including H2S. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has important implications for the winemaking industry where a better understanding of the nutritional requirements of Saccharomyces is necessary to reduce fermentation problems and to improve final product quality. PMID- 12588543 TI - Stimulation of ligninolytic enzyme production in Phanerochaete chrysosporium by polyoxyalkanes. AB - AIMS: Poly(ethylene glycol) (PEG) and some substances similar to PEG in chemical structure were tested as stimulators of ligninolytic enzyme production in shaken culture of Phanerochaete chrysosporium. METHODS AND RESULTS: The substances that caused high enzymatic activity were linear polymers [poly(ethylene glycol), poly(propylene glycol), poly(butylene glycol) and poly(vinyl alcohol)] and cyclic polymers (crown ether). They can have terminal groups other than -OH [PEG (di)methyl ether, PEG sulphate, PEG derivative with the amino group and xanthate]. The maximum lignin peroxidase activities were compared with the surface pressure caused by the stimulator. Addition of polymers composed of charged monomer units did not increase the enzymatic activity and the fungi did not grow at all on addition of polymers having a fixed positive charge. CONCLUSIONS: Lignin peroxidase activity was increased after the addition of polymers with uncharged monomer units. It was higher and its maximum was reached in a shorter time on addition of polymers with higher molecular weights. SIGNIFICANCE AND IMPACT OF STUDY: Beside Tweens there are several polymers that stimulate ligninolytic enzyme production in shaken culture of P. chrysosporium. Their characteristics are: similarity to PEG in chemical structure, having uncharged monomer units and high molecular weight. PMID- 12588544 TI - Identification of polyhydroxyalkanoate (PHA)-producing Bacillus spp. using the polymerase chain reaction (PCR). AB - AIMS: The aim of the work was to develop efficient method to identify polyhydroxyalkanoate (PHA)-producing species of Bacillus from numerous soil isolates of bacteria. Identification of the isolates and characterization of the PHA produced by strains positive on the polymerase chain reaction (PCR) was envisaged. METHODS AND RESULTS: Different bacteria isolated from soil were screened by PCR using two sets of primers designed for Bacillus megaterium. Amongst 23 isolates examined, the DNA of 12 isolates reacted positively with the primers giving amplicons identical in size to that obtained from B. megaterium. The isolates which were identified as strains of B. sphaericus, B. circulans, B. brevis and B. licheniformis, produced 11- 41% of PHA in biomass, in sucrose containing medium, over a growth period of 24-72 h. The nature of the PHA thus produced was analyzed by Fourier transform infrared spectroscopy, gas chromatography and by nuclear magnetic resonance (NMR) and found to contain polyhydroxy butyrate and polyhydroxyvalerate. CONCLUSIONS: The results indicate that most of our isolates from different species contained the B. megaterium type of PHA synthase. Bacillus licheniformis appeared to belong to another group as it did not react with both sets of primers. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the universality of the B. megaterium type of PHA synthase in soil isolates of Bacillus. Some variations were also found. PMID- 12588545 TI - Expression of a Bacillus thuringiensisdelta-endotoxin cry1Ab gene in Bacillus subtilis and Bacillus licheniformis strains that naturally colonize the phylloplane of tomato plants (Lycopersicon esculentum, Mills). AB - AIMS: To introduce a cry gene into microorganisms that naturally colonize the phylloplane of tomato plants to improve the persistence of the Cry proteins for controlling a South American tomato moth (Tuta absoluta, Meyrick, 1917). METHODS AND RESULTS: A cry1Ab gene isolated from a native Bacillus thuringiensis strain (LM-466), showing a relevant activity against T. absoluta larvae, was cloned into the shuttle vector pHT315 (Arantes and Lereclus 1991). The construct was introduced by electroporation into native Bacillus subtilis and Bacillus licheniformis strains, both natural inhabitants of the tomato phylloplane. Western analysis and toxicity assays against the target larvae proved that the successful expression of the gene was accomplished in host bacteria. Recombinant toxin displayed a similar LC50 value in comparison to native donor strain LM-466. Both transformed Bacillus survived for at least 45 days on the tomato leaf surface. CONCLUSIONS: Plant-associated microorganisms that naturally colonize the phylloplane could be useful as recombinant microbial delivery systems of toxin genes of B. thuringiensis. SIGNIFICANCE AND IMPACT OF THE STUDY: Modified microorganisms capable of surviving on leaf surfaces for several weeks with insecticidal activity should allow for a reduction in pesticide application. PMID- 12588546 TI - Evaluation of a DNA probe test kit for detection of Salmonellae in biosolids. AB - AIMS: Current US regulations (40 CFR 503) for 'Class A' biosolids (treated sewage sludge) requires use of multiple-tube fermentation techniques for fecal coliform or multiple tube enrichment techniques for Salmonella spp. followed by isolation and biochemical and serologic confirmation. The technical difficulties and the time required to complete the procedure for enumeration of Salmonellae in biosolids and sludges has limited the use of this assay. This study was conducted to determine if a commercially available molecular probe system could be used to isolate and enumerate Salmonella spp. in biosolids or sludges in less time than cultural techniques with biochemical confirmation. METHODS AND RESULTS: Several types of treated and untreated municipal sludges were assayed for Salmonellae using a cultural technique with biochemical and serologic confirmation and a DNA probe diagnostic test kit. The results indicate that the molecular probe and the conventional fermentation tube technique yielded equivalent results. Interestingly, the probe technique yielded results within 52 h following initiation of sample analysis compared with the conventional fermentation tube technique with confirmation which required approx. 120 h. CONCLUSIONS: These results suggest that the molecular probe system used for this work may be used to determine the presence or absence of Salmonella spp. in biosolids within a relatively short time frame. SIGNIFICANCE AND IMPACT OF THE STUDY: The ease of using the DNA probe test kit, along with its ability to produce results in less than half the time of conventional culture techniques, suggests that this assay is useful for determining the presence or absence of Salmonellae in biosolids samples. PMID- 12588547 TI - Properties of polysaccharide produced by Azotobacter vinelandii cultured on 4 hydroxybenzoic acid. AB - AIMS: Characterization of the exopolysaccharide produced by Azotobacter vinelandii grown on 4-hydroxybenzoic acid (EPS I), and the comparison between this exopolysaccharide and commercial alginate, constituted the main objective of this work. METHODS AND RESULTS: Total carbohydrates, uronic acids, acetyl and pyruvyl groups and proteins were determined by colorimetric methods and composition was confirmed by Nuclear Magnetic Resonance studies. Rheological properties were analysed under different physical and chemical conditions. Results showed differences between EPS I and commercial alginate, in relation to both composition and viscosity. Higher amount of guluronnosyl residues were found in EPS I, whereas commercial alginate contained the same proportion of mannuronosyl and guluronnosyl residues. In accordance with this result, EPS I gave rise to solutions of higher viscosity than commercial alginate, although solutions of this polysaccharide showed greater stability when conditions were altered. CONCLUSIONS: The exopolysaccharide produced by A. vinelandii grown on 4 hydroxybenzoic acid showed a different composition in comparison with commercial alginate, which leads to higher viscosity values for the aqueous solutions of EPS I. SIGNIFICANCE AND IMPACT OF STUDY: This work describes for the first time the characteristics of an exopolysaccharide produced by A. vinelandii from 4 hydroxybenzoic acid, a substrate rarely used as sole carbon source. PMID- 12588548 TI - Degradation of proteins during the fermentation of African locust bean (Parkia biglobosa) by strains of Bacillus subtilis and Bacillus pumilus for production of Soumbala. AB - AIMS: To examine isolates of Bacillus subtilis and B. pumilus predominant in Soumbala for their ability to degrade African locust bean proteins (ALBP). METHODS AND RESULTS: Agar diffusion test in casein and ALBP agar was used for screening of isolates. The profiles of water-soluble proteins and free amino acids (FAA) during the fermentation of ALBP by the Bacillus isolates were studied by SDS-PAGE and cation exchange chromatography. The profile of soluble proteins changed with the fermentation time and varied depending on the isolate. The quantity of total FAA and essential FAA such as lysine was increased sharply between 24 and 48 h of fermentation and differed among the isolates. Simultaneously, a pH increase was observed. Cysteine, methionine, leucine, isoleucine, tyrosine and phenylalaline appeared during fermentation. CONCLUSION: The Bacillus isolates studied degraded ALBP leading to a profile of soluble proteins and FAA specific for each isolate. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to the selection of Bacillus strains to be used as starter cultures for controlled production of Soumbala. PMID- 12588549 TI - Characterization of intestinal lactobacilli as putative probiotic candidates. AB - AIMS: To use antioxidative activity and antagonistic properties of lactobacilli against selected pathogens and members of the normal microflora as a basis for screening probiotic candidates. METHODS AND RESULTS: Antagonistic activity of lactobacilli against target bacteria in both microaerobic and anaerobic environments was tested. Production of antagonistic metabolites (ethanol, hydrogen peroxide (H2O2), acetic, lactic and succinic acid) by lactobacilli as well as their total antioxidative activity were assessed. In general, the lactobacilli tested were most effective against Gram-negative bacteria and their antagonistic activity was strain-specific. However, obligately heterofermentative lactobacilli had the strongest activity when tested in a microaerobic environment. Additionally, facultatively heterofermentative lactobacilli were equally effective in either milieu and produced significant levels of acetic and lactic acid. Moreover, obligately homofermentative lactobacilli had high H2O2 production and total antioxidative activity but weak antagonistic activity. CONCLUSIONS: Antioxidative and antagonistic activity of intestinal lactobacilli is strain-specific but typically can be related to their fermentation type which may be used for rapidly screening large numbers of lactobacilli for probiotic candidates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents the first report on the utilization of group characteristics to screen lactobacilli intended for specific probiotic use. Such uses include the targeting of particular gut niches and pathogens as well as allowing for long-term benefits to the host. PMID- 12588550 TI - Comparison of adhesion of wound isolates of Staphylococcus aureus to immobilized proteins. AB - AIMS: To determine the ability of 149 clinical isolates of Staphylococcus aureus from burns, other wounds and environmental isolates to adhere to immobilized proteins. METHODS AND RESULTS: The ability to bind to immobilized fibrinogen, fibronectin, laminin, collagen, IgG and lysozyme was studied using a microtitre plate assay. The strains were very diverse. Binding to fibrinogen was most frequent, followed by fibronectin, collagen and laminin. Binding to IgG and lysozyme was weak and few strains showed strong binding. Numerical analysis showed that 65% of the strains infecting burns had similar properties and bound to fibrinogen, fibronectin, collagen and IgG. The strains infecting other wounds had more variable characteristics. CONCLUSIONS: The ability to adhere to proteins is important in wound infection, but clinical isolates were diverse in their ability to bind to the proteins tested. Burn wounds were more likely to be infected with strains showing multiple binding characteristics. SIGNIFICANCE AND IMPACT OF THE STUDY: The study confirms the importance of adhesins in clinical infection. PMID- 12588551 TI - Detection and quantification of Escherichia coli O157:H7 in environmental samples by real-time PCR. AB - AIMS: To apply the real-time Polymerase chain reaction (PCR) method to detect and quantify Escherichia coli O157:H7 in soil, manure, faeces and dairy waste washwater. METHODS AND RESULTS: Soil samples were spiked with E. coli O157:H7 and subjected to a single enrichment step prior to multiplex PCR. Other environmental samples suspected of harbouring E.coli O157:H7 were also analysed. The sensitivity of the primers was confirmed with DNA from E.coli O157:H7 strain 3081 spiked into soil by multiplex PCR assay. A linear relationship was measured between the fluorescence threshold cycle (C T ) value and colony counts (CFU ml( 1)) in spiked soil and other environmental samples. The detection limit for E.coli O157:H7 in the real-time PCR assay was 3.5 x 10(3) CFU ml(-1) in pure culture and 2.6 x 10(4) CFU g(-1) in the environmental samples. Use of a 16-h enrichment step for spiked samples enabled detection of <10 CFU g(-1) soil. E. coli colony counts as determined by the real-time PCR assay, were in the range of 2.0 x 10(2) to 6.0 x 10(5) CFU PCR (-1) in manure, faeces and waste washwater. CONCLUSIONS: The real-time PCR-based assay enabled sensitive and rapid quantification of E. coli O157:H7 in soil and other environmental samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to quantitatively determine cell counts of E.coli O157:H7 in large numbers of environmental samples, represents considerable advancement in the area of pathogen quantification for risk assessment and transport studies. PMID- 12588552 TI - Transcription profile of brewery yeast under fermentation conditions. AB - AIMS: Yeast strains, used in the brewing industry, experience distinctive physiological conditions. During a brewing fermentation, yeast are exposed to anaerobic conditions, high pressure, high specific gravity and low temperatures. The purpose of this study was to examine the global gene expression profile of yeast subjected to brewing stress. METHODS AND RESULTS: We have carried out a microarray analysis of a typical brewer's yeast during the course of an 8-day fermentation in 15 degrees P wort. We used the probes derived from Saccharomyces cerevisiae genomic DNA on the chip and RNA isolated from three stages of brewing. This analysis shows a high level of expression of genes involved in fatty acid and ergosterol biosynthesis early in fermentation. Furthermore, genes involved in respiration and mitochondrial protein synthesis also show higher levels of expression. CONCLUSIONS: Surprisingly, we observed a complete repression of many stress response genes and genes involved in protein synthesis throughout the 8 day period compared with that at the start of fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This microarray data set provides an analysis of gene expression under brewing fermentation conditions. The data provide an insight into the various metabolic processes altered or activated by brewing conditions of growth. This study leads to future experiments whereby selective alterations in brewing conditions could be introduced to take advantage of the changing transcript profile to improve the quality of the brew. PMID- 12588553 TI - Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract. AB - AIMS: The study of two human strains of Lactobacillus to be used as probiotics in the gastrointestinal tract. METHODS AND RESULTS: The Lactobacillus acidophilus UO 001 and Lact. gasseri UO 002, were resistant to the gastrointestinal conditions (pH 2 and 3, presence of pepsin, pancreatin or bile salts), the resistance was enhanced in the presence of skimmed milk. Additionally, adhered to Caco-2 cells through glycoproteins in Lact. gasseri and carbohydrates in the case of Lact. acidophilus. These strains are able to inhibit the growth of certain enteropathogens: Salmonella, Listeria and Campylobacter without interfering with the normal microbiota of the gastrointestinal tract, as stated by using the mixed culture and the spot agar test. Finally, strongly adherent Lact. gasseri were found to inhibit the attachment of Escherichia coli O111 to intestinal Caco-2 cells under the condition of exclusion. CONCLUSIONS: These results indicate that the two strains of Lactobacillus from human origin present important properties for survival in, and colonization of, the gastrointestinal tract, that give them potential probiotic. SIGNIFICANCE AND IMPACT OF THE STUDY: Two strains of Lactobacillus isolated from human vagina of healthy premenopausal women could be promising candidates to be used in the preparation of probiotic products and for their use as health-promoting bacteria. PMID- 12588554 TI - Moderation of oral bacterial adhesion on saliva-coated hydroxyapatite by polyaspartate. AB - AIMS: Synthetic sodium alpha,beta-polyaspartate (PA) has been investigated as a moderator of adhesion and the subsequent biofilm formation by oral bacteria. METHODS AND RESULTS: The inhibition of bacterial adhesion by PA was assessed by (i) a 30-min incubation with Streptococcus sanguis in a microtitre assay with the wells coated with hydroxyapatite (HAP) and (ii) an 18-h challenge with human salivary microflora in a HAP disc assay. In contrast to HAP-coated surfaces, clean polystyrene surfaces in the microtitre assay exhibited no anti-adhesion properties. It has been found that PA significantly and similarly adsorbs onto HAP surfaces in the presence and absence of salivary coating. The HAP disc assay also showed that PA, both in aqueous solutions and in toothpaste, reduced the level of adhered microflora and this effect was enhanced by added propylene oxide ethylene oxide copolymers. CONCLUSION: The principal finding from this work is the potential role for PA as an inhibitor of dental plaque formation. PA may significantly modify the salivary pellicle. SIGNIFICANCE AND IMPACT OF THE STUDY: This work indicates the use of PA in controlling the development of dental plaque and the formation of bacterial biofilm in general. PMID- 12588555 TI - Volatile compounds produced by Lactobacillus fermentum, Saccharomyces cerevisiae and Candida krusei in single starter culture fermentations of Ghanaian maize dough. AB - AIMS: To identify and compare the volatile compounds associated with maize dough samples prepared by spontaneous fermentation and by the use of added starter cultures in Ghana. METHODS AND RESULTS: The starter cultures examined were Lactobacillus fermentum, Saccharomyces cerevisiae and Candida krusei. For identification of aroma volatiles, extracts by the Likens-Nickerson simultaneous distillation and extraction technique were analysed by gas chromatography-mass spectrometry (GC-MS) and using a trained panel of four judges by GC-Olfactometry (GC-sniffing). Compounds identified by GC-MS in maize dough samples after 72 h of fermentation included 20 alcohols, 22 carbonyls, 11 esters, seven acids, a furan and three phenolic compounds. Of the total 64 volatile compounds, 51 were detected by GC-sniffing as contributing to the aroma of the different fermented dough samples. Spontaneously fermented maize dough was characterized by higher levels of carbonyl compounds while fermentations with added L. fermentum recorded the highest concentration of acetic acid. S. cerevisiae produced higher amounts of fusel alcohols and increasing levels of esters with fermentation time and C. krusei showed similarity to L. fermentum with lower levels of most volatiles identified. CONCLUSION: The present study has given a detailed picture of the aroma compounds in fermented maize and demonstrated that the predominant micro organisms in fermented maize dough can be used as starter cultures to modify the aroma of fermented maize dough. SIGNIFICANCE AND IMPACT OF THE STUDY: The study has documented the advantage of using starter cultures in African traditional food processing and provided a scientific background for introducing better controlled fermentations. PMID- 12588556 TI - Temperature effect on Fusarium oxysporum f.sp. melonis survival during horticultural waste composting. AB - AIMS: The aim of this work was to study the effect of high temperatures generated during composting process, on the phytopathogen fungus Fusarium oxysporum f.sp. melonis. This investigation was achieved by both in vivo (semipilot-scale composting of horticultural wastes) and in vitro (lab-scale thermal treatments) assays. METHODS AND RESULTS: Vegetable residues infected with F. oxysporum f.sp. melonis were included in compost piles. Studies were conducted in several compost windrows subjected to different treatments. Results showed an effective suppression of persistence and infective capacity, as this process caused complete fungal elimination after 2-3 days of composting. In order to confirm the effect of high temperature during this process, in vitro experiments were carried out. Temperature values of 45, 55 and 65 degrees C were tested. All three treatments caused the elimination of fungal persistence. Treatment at 65 degrees C was especially effective, whereas 45 degrees C eliminated fungal persistence only after 10 days. CONCLUSIONS: The composting process is an excellent alternative for the management of plant wastes after harvesting, as this procedure is able to suppress infective capacity of several harmful phytopathogens such as F. oxysporum f.sp. melonis. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium oxysporum f.sp. melonis is a plant pathogen fungus specially important in the province of Almeria (south-east Spain), where intensive greenhouse horticulture is very extended. High temperatures reached during composting of horticultural plant wastes ensure the elimination of phytopathogen microorganisms such as F. oxysporum f.sp. melonis from vegetable material, providing an adequate hygienic quality in composts obtained. PMID- 12588557 TI - Image analysis method for evaluation of specific and non-specific hand contamination. AB - AIMS: To evaluate a quantifying image analysis method for assessing the degree of hand contamination and efficacy of hand washing procedures. METHODS AND RESULTS: Two types of experimental design were used. In one, different concentrations of pure cultures of Escherichia coli, Listeria innocua and Pseudomonas flourescens were applied to hands. In the other, hands were contaminated by handling various raw foods. Imprints of the contaminated palms were made on 24.5 x 24.5 cm agar plates using appropriate agars. After incubation, digital photographs of the plates were analysed using image analysis. In pure culture studies with selective agars, levels from 1 to 10(6) CFU cm(-2) palm could be monitored. For aerobic, mesophilic organisms from raw chicken, levels from 10(3) to 10(6) CFU cm(-2) palm were correlated linearly to image analysis data. CONCLUSIONS: The image analysis of palm imprints made on agar plates was suitable for assessing the degree of contamination from foods on the palms. Sensitivity and specificity depended on the agar used and the type of contamination encountered. SIGNIFICANCE AND IMPACT OF THE STUDY: Data capture by the image analysis method is simple and can be partly automated. Sampling time is short for the person to be tested, which makes it an attractive method for assessing hand hygiene status in larger field trials. PMID- 12588558 TI - Antimicrobial resistance and genomic screening of clinical isolates of thermophilic Campylobacter spp. from south-east Queensland, Australia. AB - AIM: To screen 90 clinical isolates of thermophilic Campylobacter species for putative resistance to ampicillin, erythromycin and tetracycline and perform numerical analysis to determine isolate relatedness. METHODS AND RESULTS: Disc diffusion, E-test MIC and agar dilution methods were performed. Disc diffusion testing showed 87 (97%) isolates appeared resistant to ampicillin at 10 microg; 14 (16%) resistant to tetracycline at 30 microg; and three (3.4%) resistant to erythromycin at 15 microg. E-test MICs showed a range of 0.5 to >256 mg l(-1) for ampicillin; 16 to >256 mg l(-1) for tetracycline; and >256 mg l(-1) for erythromycin. E-test showed 68% correlation (+/-1 log2 dilution) with agar dilution for ampicillin, 100% for erythromycin and 64% for tetracycline. Disc diffusion testing showed 100% correlation with agar dilution for erythromycin and tetracycline, and 77% for ampicillin. Numerical analyses of restriction endonuclease (RE) fragment profiles suggested a high level of isolate variation. CONCLUSION: The incidence of resistance of thermophilic Campylobacter spp. to erythromycin and tetracycline is low in south-east Queensland. SIGNIFICANCE AND IMPACT OF THE STUDY: Disc diffusion susceptibility testing may be used to screen thermophilic Campylobacter spp. for putative resistance to erythromycin and tetracycline. Agar dilution should be used to determine ampicillin susceptibility. PMID- 12588559 TI - The influence of the chemical composition of drinking water on cuprosolvency by biofilm bacteria. AB - AIMS: This study investigated the influence of water chemistry on copper solvation (cuprosolvency) by pure culture biofilms of heterotrophic bacteria isolated from copper plumbing. METHODS AND RESULTS: Heterotrophic bacteria isolated from copper plumbing biofilms including Acidovorax delafieldii, Flavobacterium sp., Corynebacterium sp., Pseudomonas sp. and Stenotrophomonas maltophilia were used in laboratory coupon experiments to assess their potential for cuprosolvency. Sterile copper coupons were exposed to pure cultures of bacteria to allow biofilm formation and suspended in drinking waters with different chemical compositions. Sterile coupons not exposed to bacteria were used as controls. After 5 days of incubation, copper release and biofilm accumulation was quantified. The results demonstrated that cuprosolvency in the control experiments was influenced by water pH, total organic carbon (TOC) and conductivity. Cuprosolvency in the presence of biofilms correlated with the chemical composition of the water supplies particularly pH, Langeliers Index, chloride, alkalinity, TOC and soluble phosphate concentrations. CONCLUSIONS: The results suggest water quality may influence cuprosolvency by biofilms present within copper plumbing pipes. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential for water chemistry to influence cuprosolvency by biofilms may contribute to the sporadic nature of copper corrosion problems in distribution systems. PMID- 12588560 TI - Drying of Epicoccum nigrum conidia for obtaining a shelf-stable biological product against brown rot disease. AB - AIMS: The effects of freeze-drying, spray-drying and fluidized bed-drying on survival of Epicoccum nigrum conidia were compared. METHODS AND RESULTS: Viability of E. nigrum conidia (estimated by measuring its germination) was 100% after fluidized bed-drying and freeze-drying, but it was determined that skimmed milk must be added in the case of freeze-drying conidia. Addition of other protectants (Tween-20, peptone, sucrose, glucose, starch and peptone + starch) to skimmed milk before freeze-drying did not improve the conidial viability which was obtained with skimmed milk alone. Glycerol had a negative effect on the lyophilization of E. nigrum conidia. Epicoccum nigrum conidia freeze-dried with skimmed milk, or fluidized bed-dried alone maintained an initial viability for 30 and 90 days, respectively, for storage at room temperature. Epicoccum nigrum conidial viability after spray-drying was lower than 10%. CONCLUSIONS: The best method to dry E. nigrum conidia was fluidized bed-drying. Conidia without protectants dried by this method had 100% viability and survived for 90 days at room temperature. SIGNIFICANCE AND IMPACT OF STUDY: This paper deals with methods for the potential formulation of a biocontrol agent which is being tested for eventual commercialization. PMID- 12588561 TI - PCR detection of psychrophilic Clostridium spp. causing 'blown pack' spoilage of vacuum-packed chilled meats. AB - AIMS: To develop a practical molecular procedure that directly, without isolation, and specifically detects the presence of clostridia which cause 'blown pack' spoilage of vacuum-packed meat. METHODS AND RESULTS: Primer sets and PCR amplification procedures were developed that detect the presence of 16S rDNA gene and/or 16S-23S rDNA internal transcribed spacer fragments of 'blown pack' causing clostridia in meat. The specificity of the developed procedures was evaluated with DNA obtained from close phylogenetic neighbours of 'blown pack' causing clostridia, food clostridia and common meat spoilage microorganisms. The sensitivity of detection was assessed in non-enriched and low-temperature enriched beef mince inoculated with serially diluted pure cultures of Clostridium estertheticum DSMZ 8809T and Cl. gasigenes DB1AT. The efficacy of detection procedures was evaluated for naturally contaminated vacuum-packed meat samples. Three primer sets, 16SE, 16SDB and EISR, produced amplicons of the expected size with DNA templates from target clostridia, but failed to yield PCR products with DNAs from any other microorganisms tested. With 16SE and 16SDB primers, minimum levels of detection were 104 CFU g(-1) for non-enriched, and 102 CFU g(-1) for enriched meat samples. Based on the established specificity of these primers, as well as DNA sequencing of amplicons, Cl. gasigenes was confirmed as the causative agent of 'blown pack' spoilage in two packs, and Cl. estertheticum as the causative agent in the third. CONCLUSIONS: The developed method can be used for rapid detection of 'blown pack' causing clostridia in commercial blown packs, or following low temperature enrichment, for detection of these microorganisms in meat containing as few as 100 clostridial cells per gram. SIGNIFICANCE AND IMPACT OF THE STUDY: The paper reports practical procedures that can be used for rapid confirmation of the causative agents of clostridial 'blown pack' spoilage in commercial spoiled packs, or for detection of psychrophilic clostridia in epidemiological trace back of 'blown pack' spoilage incidents in meat processing plants. PMID- 12588562 TI - Partial characterization of bacteriocins produced by environmental strain Enterococcus faecium EK13. AB - AIMS: The partial characterization of bacteriocins produced by an environmental strain Enterococcus faecium EK13, isolated from cattle dung water. METHODS AND RESULTS: A bacteriocin was partially purified by ammonium sulphate precipitation, followed by a SP-Sepharose column, reverse-phase chromatography and N-terminal region sequenced. The anti-microbial substance produced was found to be a heat stable polypeptide with molecular mass 4.83 kDa, which was determined by N terminal amino acid sequencing to be enterocin A. A second substance was specified by PCR as enterocin P. Bacteriocins were stable at 4 and -20 degrees C for long storage periods. The optimum of bacteriocin production was observed in the range of pH 5.0-6.5 at 30 and 37 degrees C. The most active substances are produced by strain EK13 in logarithmic growth phase and bacteriocins are produced after 1 h of fermentation. The highest activity detected in fermentation experiments was 51 200 AU ml(-1) and the most sensitive indicator strain was found to be Listeria innocua LMG 13568. Differences in bacteriocin activity against two indicators could be explained by more than one type of enterocin production by strain EK13, or with different mode of action or in different sensitivity of strains. CONCLUSION: Enterococcus faecium strain EK13 isolated from cattle dung water produces two bacteriocins, enterocin A and P, with an inhibitory effect against the strain of the genera Enterococcus, Leuconostoc, Lactobacillus, Streptococcus, Staphylococcus, Bacillus and Listeria (in different origin). SIGNIFICANCE AND IMPACT OF THE STUDY: Enterococcus faecium EK13 environmental strain is a new producer of enterocin A and P. The E. faecium EK13, isolated from cattle dung water, is presented with the further aim to utilize it for waste treatment by biotechnological processes. PMID- 12588563 TI - Solar inactivation of mesophilic Aeromonas by exogenous photooxidation in high rate algal pond treating waste water. AB - AIMS: Investigations were carried out to observe the effect of sunlight on the survival of mesophilic Aeromonas (A. caviae, A. hydrophila and A. sobria) in high rate algal pond. METHODS AND RESULTS: Light damage was estimated by loss of bacterial culturability, using simple beaker experiments. Survival of the strains studied under illuminated conditions was highly affected by the physico-chemical conditions within the microcosms. The inactivation of Aeromonas increased strongly as dissolved oxygen was increased, and was dependent on exogenous sensitizers. This process was enhanced by the high pH of water. Die-off of bacteria was prevented by adding catalase and pyruvate (H2O2 scavengers) into the microcosms. CONCLUSIONS: The damage of Aeromonas was mainly due to exogenous photooxidation. Hydrogen peroxide at least was involved in light-induced damage of bacteria. Aeromonas sobria appeared to be slightly less sensitive to photooxidation than A. hydrophila and A. caviae. SIGNIFICANCE AND IMPACT OF STUDY: The present study could explain the relative resistance of A. sobria to treatment in wastewater stabilization ponds. PMID- 12588564 TI - Anemia in the elderly: clinical impact and practical diagnosis. PMID- 12588565 TI - Epidemiology of anemia in the elderly: information on diagnostic evaluation. AB - A rise in the aging population has been predicted, and, as a result, it is expected that the incidence of age-related health conditions will also increase. Although common in the elderly, anemia is often mild and asymptomatic and rarely requires hospitalization. However, untreated anemia can be detrimental, because it is associated with increased mortality, poor health, fatigue, and functional dependence and can lead to cardiovascular and neurological complications. Several factors have been suggested to cause anemia in this population, for example, blood loss or chronic disease. In some cases, the cause is unknown. It has been suggested that this is a result of the presence of comorbid conditions that can mask the symptoms of anemia. Therefore, appropriate diagnosis and management strategies of anemia in the elderly need to be identified, particularly because anemia may indicate the presence of other serious diseases. PMID- 12588566 TI - Medical and functional consequences of anemia in the elderly. AB - Anemia is a common problem in the elderly, accounting for significant morbidity and mortality in this population. It also has a negative effect on quality of life. Recent findings have shown that anemia can lead to cardiovascular and neurological complications, such as congestive heart failure and impaired cognitive function. In addition, anemia has been implicated in functional impairment and falls. Available data have shown that the successful management of this condition will not only improve patients' quality of life, but may also prevent the anemia from worsening. PMID- 12588567 TI - Cost of anemia in the elderly. AB - As the aging population and the incidence of age-related health conditions increase, the cost of healthcare is also expected to rise. Anemia commonly occurs in the elderly, and is associated with a number of health conditions such as falls, weakness, and immobility. It can also lead to more-serious complications such as cardiovascular and neurological impairments. Consequently, anemia can have a significant effect on healthcare requirements and healthcare expenditure. Research is needed into the opportunity to reduce costs. PMID- 12588568 TI - Biological interactions of aging and anemia: a focus on cytokines. AB - Anemia is one of the characteristics of the frailty phenotype and is often observed in elderly patients. Although anemia in people of advancing age can often be attributed to underlying etiologies such as iron deficiency or chronic disease, some cases do not have any identifiable cause. Therefore, it has been suggested that the aging process itself might be an intrinsic factor in the development of anemia, possibly through the age-related dysregulation of certain proinflammatory cytokines such as interleukin-6 (IL-6). Although the mechanism underlying the association between increased IL-6 and anemia has not been fully elucidated, it has been suggested that, like with other cytokines, it involves direct inhibition of erythropoietin production or interaction with the erythropoietin receptor. PMID- 12588569 TI - Disordered hematopoiesis and myelodysplasia in the elderly. AB - Normal hematopoiesis constitutes the process of producing diverse, differentiated blood cell types in a manner related to physiological requirement. During aging, modulation of hematopoiesis becomes disordered, impairing the ability of older people to respond appropriately to the physiological demand for blood cell replacement triggered by stimuli such as blood loss or cytoreductive chemotherapy. This may contribute to the increase in the prevalence of anemia that is observed during aging. In addition, various age-related events, such as genomic mutations secondary to oxidative stress and impaired regulation of cytokine production, may contribute to or cause the emergence of abnormal clones of hematopoietic cells. Therefore, normal hematopoiesis is disrupted, and the hematopoietic system is populated with cells that are quantitatively and functionally deficient and are also subject to leukemic transformation. These defects in the production and maturation of the various differentiated blood cells are referred to as myelodysplastic syndromes. These syndromes are so tightly associated with aging that they are considered to be geriatric disorders; they can lead to anemia, neutropenia, and thrombocytopenia and to the development of acute nonlymphoblastic leukemia. Dysregulation of mechanisms controlling hematopoiesis is therefore an important characteristic of the hematopoietic system in the elderly, but the response of progenitor cells to humoral stimulators is preserved and accounts for the effectiveness of recombinant hematopoietic growth factors used as emerging treatment modalities for hematopoietic disorders in the elderly. PMID- 12588571 TI - A randomized, controlled trial of quadriceps resistance exercise and vitamin D in frail older people: the Frailty Interventions Trial in Elderly Subjects (FITNESS). AB - OBJECTIVES: To determine the effectiveness of vitamin D and home-based quadriceps resistance exercise on reducing falls and improving the physical health of frail older people after hospital discharge. DESIGN: Multicenter, randomized, controlled trial with a factorial design. SETTING: Five hospitals in Auckland, New Zealand, and Sydney, Australia. PARTICIPANTS: Two hundred forty-three frail older people. INTERVENTIONS: Patients were randomized to receive a single dose of vitamin D (calciferol, 300,000 IU) or placebo tablets and 10 weeks of high intensity home-based quadriceps resistance exercise or frequency-matched visits. MEASUREMENTS: The primary endpoints were physical health according to the short form health survey at 3 months and falls over 6 months. Physical performance and self-rated function were secondary endpoints. Assessments took place in the participants' homes at 3 and 6 months after randomization and were performed by blinded assessors. RESULTS: There was no effect of either intervention on physical health or falls, but patients in the exercise group were at increased risk of musculoskeletal injury (risk ratio = 3.6, 95% confidence interval = 1.5 8.0). Vitamin D supplementation did not improve physical performance, even in those who were vitamin D deficient (<12 ng/mL) at baseline. CONCLUSION: Neither vitamin D supplementation nor a home-based program of high-intensity quadriceps resistance exercise improved rehabilitation outcomes in frail older people after hospitalization. There was no effect of vitamin D on physical performance, and the exercises increased the risk of musculoskeletal injury. These findings do not support the routine use of these interventions at these dosages in the rehabilitation of frail older people. PMID- 12588572 TI - Preventing falls in community-dwelling frail older people using a home intervention team (HIT): results from the randomized Falls-HIT trial. AB - OBJECTIVES: To evaluate the effect of an intervention by a multidisciplinary team to reduce falls in older people's homes. DESIGN: Randomized, controlled trial with follow-up of subjects for 1 year. SETTING: University-affiliated geriatric hospital and older patients' homes. PARTICIPANTS: Three hundred sixty subjects (mean age +/- standard deviation = 81.5 +/- 6.4) admitted from home to a geriatric hospital and showing functional decline, especially in mobility. INTERVENTION: The participants were randomly assigned to receive a comprehensive geriatric assessment followed by a diagnostic home visit and home intervention or a comprehensive geriatric assessment with recommendations and usual care at home. The home intervention included a diagnostic home visit, assessing the home for environmental hazards, advice about possible changes, offer of facilities for any necessary home modifications, and training in the use of technical and mobility aids. An additional home visit was made after 3 months to reinforce the recommendations. After 12 months of follow-up, a home visit was made to all study participants. MEASUREMENTS: Number of falls, type of recommended home modifications, and compliance with recommendations. RESULTS: After 1 year, there were 163 falls in the intervention group and 204 falls in the control group. The intervention group had 31% fewer falls than the control group (incidence rate ratio (IRR) = 0.69, 95% confidence interval (CI) = 0.51-0.97). The intervention was most effective in a subgroup of participants who reported having had two or more falls during the year before recruitment into the study. In this subgroup, the proportion of frequent fallers and the rate of falls was significantly reduced for the intervention group compared with the control group (21 vs 36 subjects with recurrent falls, P =.009; IRR = 0.63, 95% CI = 0.43-0.94). The compliance rate varied with the type of change recommended from 83% to 33% after 12 months of follow-up. CONCLUSION: Home intervention based on home visits to assess the home for environmental hazards, providing information about possible changes, facilitating any necessary modifications, and training in the use of technical and mobility aids was effective in a selected group of frail older subjects with a history of recurrent falling. PMID- 12588573 TI - Effectiveness of a multifaceted intervention on falls in nursing home residents. AB - OBJECTIVES: To evaluate the effectiveness of a multifaceted, nonpharmaceutical intervention on incidence of falls and fallers. DESIGN: Prospective, cluster randomized, controlled 12-month trial. SETTING: Six community nursing homes in Germany. PARTICIPANTS: Long-stay residents (n = 981) aged 60 and older; mean age 85; 79% female. INTERVENTIONS: Staff and resident education on fall prevention, advice on environmental adaptations, progressive balance and resistance training, and hip protectors. MEASUREMENTS: Falls, fallers, and fractures. RESULTS: The incidence density rate of falls per 1,000 resident years (RY) was 2,558 for the control group (CG) and 1,399 for the intervention group (IG) (relative risk (RR) = 0.55, 95% confidence interval (CI) = 0.41-0.73). Two hundred forty-seven (52.3%) fallers were detected in the CG and 188 (36.9%) in the IG (RR = 0.75, 95% CI = 0.57-0.98). The incidence density rate of frequent fallers (>2/year) was 115 (24.4%) for the CG and 66 (13.0%) for the IG (RR = 0.56, 95% CI = 0.35-0.89). The incidence density rate of hip fractures per 1,000 RY was 39 for the CG and 43 for the IG (RR = 1.11, 95% CI = 0.49-2.51). Other fractures were diagnosed with an incidence density rate of 52 per 1,000 RY for CG and 41 per 1,000 RY for IG (RR = 0.78, 95% CI = 0.57-1.07). CONCLUSION: The incidence density rate of falls and fallers differed considerably between the control and intervention groups. The study was underpowered to demonstrate a significant difference of hip or nonhip fractures. Because of a low fracture rate in both groups, the investigation of fracture rates would have required a larger sample size to detect an effect of the intervention. PMID- 12588574 TI - Physical performance measures in the clinical setting. AB - OBJECTIVES: To assess the ability of gait speed alone and a three-item lower extremity performance battery to predict 12-month rates of hospitalization, decline in health, and decline in function in primary care settings serving older adults. DESIGN: Prospective cohort study. SETTING: Primary care programs of a Medicare health maintenance organization (HMO) and Veterans Affairs (VA) system. PARTICIPANTS: Four hundred eighty-seven persons aged 65 and older. MEASUREMENTS: Lower extremity performance Established Population for Epidemiologic Studies of the Elderly (EPESE) battery including gait speed, chair stands, and tandem balance tests; demographics; health care use; health status; functional status; probability of repeated admission scale (Pra); and primary physician's hospitalization risk estimate. RESULTS: Veterans had poorer health and higher use than HMO members. Gait speed alone and the EPESE battery predicted hospitalization; 41% (21/51) of slow walkers (gait speed <0.6 m/s) were hospitalized at least once, compared with 26% (70/266) of intermediate walkers (0.6-1.0 m/s) and 11% (15/136) of fast walkers (>1.0 m/s) (P <.0001). The relationship was stronger in the HMO than in the VA. Both performance measures remained independent predictors after accounting for Pra. The EPESE battery was superior to gait speed when both Pra and primary physician's risk estimate were included. Both performance measures predicted decline in function and health status in both health systems. Performance measures, alone or in combination with self-report measures, were more able to predict outcomes than self-report alone. CONCLUSION: Gait speed and a physical performance battery are brief, quantitative estimates of future risk for hospitalization and decline in health and function in clinical populations of older adults. Physical performance measures might serve as easily accessible "vital signs" to screen older adults in clinical settings. PMID- 12588575 TI - Strength and muscle quality in a well-functioning cohort of older adults: the Health, Aging and Body Composition Study. AB - OBJECTIVES: To determine whether lower lean mass and higher fat mass have independent effects on the loss of strength and muscle quality in older adults and might explain part of the effect of age. DESIGN: Single-episode, cross sectional analyses of a cohort of subjects in the Health, Aging and Body Composition (Health ABC) Study. SETTING: Ambulatory clinic and research laboratory. PARTICIPANTS: Two thousand six hundred twenty-three men and women aged 70 to 79 from the Health ABC Study. MEASUREMENTS: Upper and lower extremity strength was measured using isokinetic (knee extension) and isometric (grip strength) dynamometers. Body composition (lean mass and fat mass) was determined by measuring lean mass of upper and lower extremities and the total body by dual energy x-ray absorptiometry. Muscle quality was ascertained by taking the ratio of strength to muscle mass for both upper and lower extremities. RESULTS: Upper and lower extremity strength and muscle quality decreased as age increased. Most of the explained variance in strength was due to differences in muscle mass, but, in those at the extremes of body fat and lower leg muscle quality, the association with body fat was independent of the effect of age. Although blacks had greater muscle strength and mass than whites, leg muscle quality tended to be lower in blacks than in whites. Upper extremity strength adjusted for lean mass and muscle quality were also associated inversely and independently with age, body fat, and black race. CONCLUSION: In this older cohort, lower strength with older age was predominantly due to a lower muscle mass. Age and body fat also had significant inverse associations with strength and muscle quality. Both preservation of lean mass and prevention of gain in fat may be important in maintaining strength and muscle quality in old age. PMID- 12588576 TI - Baseline balance and functional decline in older adults with knee pain: the Observational Arthritis Study in Seniors. AB - OBJECTIVES: To examine the cross-sectional and longitudinal effect of baseline balance, independent of multiple covariates (sociodemographic variables, comorbidities), on self-reported disability and several measures of functional performance. DESIGN: Observational Arthritis Study in Seniors was a community based longitudinal study. SETTING: Data collection was conducted at an exercise science research laboratory. PARTICIPANTS: A cohort of 245 women and 235 men, aged 65 and older, with self-reported knee pain. MEASUREMENTS: Measurements of balance, self-reported disability, functional performance, strength, sociodemographic characteristics, disease burden (including radiographic knee osteoarthritis (OA)), and functional limitations were obtained on participants at baseline and 30 months. The four outcome measures (self-reported disability, stair climb time, car time, preferred walking speed) were also collected at 15 months. RESULTS: In cross-sectional analyses, baseline balance was significantly associated with each of the four outcome measures before adjusting for baseline knee strength. After adjusting for knee strength, baseline balance remained significantly associated with all functional performance outcomes and self reported disability. In longitudinal analyses, baseline balance had a significant effect on car-time performance, whereby individuals in the lowest quartile of baseline balance scores declined in a quadratic manner from baseline to Month 30. CONCLUSION: Baseline balance, independent of strength and various other covariates, is an important component of performance for transfer and ambulatory tasks and for self-reported disability. Balance was predictive of change in performance over time only in a task that presented a significant challenge to the lateral mobility/stability of participants. Balance was not predictive of decline in ambulatory performance-based tasks or in self-reported disability. PMID- 12588577 TI - Effect of low-dose thiazide diuretics on plasma lipids: results from a double blind, randomized clinical trial in older men and women. AB - OBJECTIVES: To determine the effects of low doses of hydrochlorothiazide (HCTZ) on plasma lipids in healthy older men and women. DESIGN: Randomized, double blind, placebo-controlled clinical trial for 3 years followed by a 1 year extension without medication. SETTING: Research clinic of a nonprofit health maintenance organization. PARTICIPANTS: Two hundred five women and 115 men aged 60 to 79 with normal blood pressure, bone density within two standard deviations of mean for age, and low-density lipoprotein cholesterol (LDL-C) levels less than 190 mg/dL. INTERVENTION: Placebo or HCTZ 12.5 or 25 mg/d. MEASUREMENTS: Plasma cholesterol, LDL-C, high-density lipoprotein cholesterol, triglycerides. RESULTS: There were no significant differences in the change in the plasma lipids between placebo and either group. The mean changes +/- standard deviation in LDL-C after 3 years in men were -5.2 +/- 3.9 mg/dL (placebo), -4.6 +/- 3.7 (12.5 mg), and 1.1 +/- 3.8 (25 mg) and in women were 1.3 +/- 2.8 (placebo), -1.1 +/- 2.8 (12.5 mg), and -6.8 +/- 2.8 (25 mg). No significant differences were seen when analysis was done only for subjects who adhered to study medication. CONCLUSION: This study provides evidence that low doses of thiazide diuretics have minimal effects on cholesterol metabolism in healthy older adults. Therefore, effects on lipid levels need not limit use of thiazides for prevention of osteoporotic fractures. PMID- 12588578 TI - The effects of an exercise and incontinence intervention on skin health outcomes in nursing home residents. AB - OBJECTIVES: To examine skin health outcomes of an exercise and incontinence intervention. DESIGN: Randomized controlled trial with blinded assessments of outcomes at three points over 8 months. SETTING: Four nursing homes (NHs). PARTICIPANTS: One hundred ninety incontinent NH residents. INTERVENTION: In the intervention group, research staff provided exercise and incontinence care every 2 hours from 8:00 a.m. to 4:30 p.m. (total of four daily care episodes) 5 days a week for 32 weeks. The control group received usual care from NH staff. MEASUREMENTS: Perineal skin wetness and skin health outcomes (primarily blanchable erythema and pressure ulcers) as measured by direct assessments by research staff, urinary and fecal incontinence frequency, and percentage of behavioral observations with resident engaged in standing or walking. RESULTS: Intervention subjects were significantly better in urinary and fecal incontinence, physical activity, and skin wetness outcome measures than the control group. However, despite these improvements, differences in skin health measures were limited to the back distal perineal area, which included the sacral and trochanter regions. There was no difference between groups in the incidence rate of pressure ulcers as measured by research staff, even though those residents who improved the most on fecal incontinence showed improvement in pressure ulcers in one area. CONCLUSION: A multifaceted intervention improved four risk factors related to skin health but did not translate into significant improvements in most measures of skin health. Even if they had adequate staffing resources, NHs might not be able to improve skin health quality indicators significantly if they attempt to implement preventive interventions on all residents who are judged at risk because of their incontinence status. PMID- 12588579 TI - Visual risk factors for hip fracture in older people. AB - OBJECTIVES: To determine the association between poor vision and risk of hip fracture in the Blue Mountains Eye Study. DESIGN: Prospective population-based cohort study. SETTING: Two post code areas in the Blue Mountains, west of Sydney, Australia. PARTICIPANTS: Three thousand six hundred fifty-four community-dwelling Australians aged 49 and older. MEASUREMENTS: At baseline, subjects had an extensive eye examination, including refraction, contrast sensitivity and visual field testing, photographs of the lens and retina, and an interview. Hip fractures during the 5-year follow-up were identified by self-report and review of medical records and were radiologically confirmed. RESULTS: For 2-year follow up (17 hip fractures), the adjusted hazard ratio (HR) for risk of hip fracture in those with corrected visual acuity worse than 20/60 was 8.4 (95% confidence interval (CI) = 1.5-48.5, population attributable risk (PAR) = 27%); for presence of posterior subcapsular cataract, the adjusted HR was 5.0 (95% CI = 1.1-23.0, PAR = 24%); and for visual field loss, the adjusted HR was 5.5 (95% CI = 1.0 29.8, PAR = 55%). In those aged 75 and older, visual acuity worse than 20/60 gave an adjusted HR of 40.6 (95% CI = 5.6-292.5, PAR = 49%). Visual impairment of any type did not predict risk of hip fracture after a 2-year follow-up. CONCLUSION: Visual impairment is strongly associated with risk of hip fracture in the next 2 years but not over a longer period of time. PMID- 12588580 TI - Estimating hip fracture morbidity, mortality and costs. AB - OBJECTIVES: To estimate lifetime morbidity, mortality, and costs from hip fracture incorporating the effect of deficits in activities of daily living. DESIGN: Markov computer cohort simulation considering short- and long-term outcomes attributable to hip fractures. Data estimates were based on published literature, and costs were based primarily on Medicare reimbursement rates. SETTING: Postacute hospital facility. PARTICIPANTS: Eighty-year-old community dwellers with hip fractures. MEASUREMENTS: Life expectancy, nursing facility days, and costs. RESULTS: Hip fracture reduced life expectancy by 1.8 years or 25% compared with an age- and sex-matched general population. About 17% of remaining life was spent in a nursing facility. The lifetime attributable cost of hip fracture was $81,300, of which nearly half (44%) related to nursing facility expenses. The development of deficits in ADLs after hip fracture resulted in substantial morbidity, mortality, and costs. CONCLUSION: Hip fractures result in significant mortality, morbidity, and costs. The estimated lifetime cost for all hip fractures in the United States in 1997 likely exceeded $20 billion. These results emphasize the importance of current and future interventions to decrease the incidence of hip fracture. PMID- 12588581 TI - Predictors of health resource use by disabled older female Medicare beneficiaries living in the community. AB - OBJECTIVES: To identify specific clinical factors that could best predict resource use by disabled older women. DESIGN: Cross-sectional. SETTING: Urban community in Baltimore, Maryland. PARTICIPANTS: One thousand two community dwelling, moderately to severely disabled, female Medicare beneficiaries aged 65 and older, from the Women's Health and Aging Study I (WHAS). MEASUREMENTS: WHAS data were merged with participants' 1992-1994 Medicare claims data for the year after baseline evaluation, reflecting inpatient, outpatient, home-based, and skilled-nursing services. The independent contributions of factors hypothesized to predict health expenditures were assessed, using chi-square and regression analyses, with the logarithm of Medicare expenditures as the primary outcome. RESULTS: Demographic factors were not associated with Medicare expenditures. Factors associated with expenditures in bivariate analyses included heart disease (1.4x), chronic obstructive pulmonary disease (1.3x), diabetes mellitus (1.1x), smoking, comorbidity, and severity of disability, as well as low creatinine clearance, serum albumin, caloric expenditure, or skinfold thickness. Heart disease, diabetes mellitus, and low skinfold thickness remained significant after adjustment for other factors. CONCLUSION: Heart disease, diabetes mellitus, and low skinfold thickness are important independent predictors of 1-year Medicare expenditures by disabled older women. Many other variables that reflect disease, disability, nutrition, or personal habits have less predictive ability. Most demographic factors are not predictors of expenditures in this population. Focusing on the best predictors may facilitate more-effective risk adjustment and creation of related health policies. PMID- 12588582 TI - Predictors of alcohol-treatment seeking in a sample of older veterans in the GET SMART program. AB - OBJECTIVES: To examine the predictive value of demographic characteristics and substance abuse indicators to explain treatment seeking for substance abuse problems by older male medical patients. DESIGN: Longitudinal analysis of screening data and treatment-seeking behavior. SETTING: Inpatient medical and outpatient substance abuse treatment center. PARTICIPANTS: Participants in the study were 855 medically ill male veterans aged 55 and older, who were screened for alcohol problems during inpatient medical treatment after clinician referral. MEASUREMENTS: The CAGE alcohol screen (Cut down on your drinking, Annoyed by criticism of your drinking, Guilty about your drinking, Eye-opener), drug use, and demographic measures administered at time of screening. Predictors of treatment seeking in the sample were examined using structural equation modeling. RESULTS: Expressed interest in treatment and later attendance at a pretreatment evaluation were associated with younger age and a higher CAGE alcohol screening score. Being unmarried and using drugs in addition to alcohol were associated with treatment interest but not with evaluation attendance. In the path model tested, the effect of higher CAGE score partially explained the effect of younger age on treatment seeking. CONCLUSION: The model examined shows utility in predicting alcohol-treatment seeking in this sample. Age-related factors may deter treatment seeking by older male medical inpatients. PMID- 12588583 TI - A home-based pedometer-driven walking program to increase physical activity in older adults with osteoarthritis of the knee: a preliminary study. AB - OBJECTIVES: To determine whether a home-based pedometer-driven walking program with arthritis self-management education (Walk +) would increase physical activity, muscle strength, and functional performance in older adults with osteoarthritis (OA) of the knee as opposed to arthritis self-management education alone (EDU). DESIGN: A randomized two-by-three (group-by-time) design with participants assigned to Walk + (n = 17, mean age +/- standard deviation = 69.6 +/- 6.7) or EDU (n = 17, age = 70.8 +/- 4.7). SETTING: Community located in the Baltimore-Washington area. PARTICIPANTS: Thirty-four community-dwelling adults, aged 60 and older with symptomatic knee OA and self-reported functional impairment. INTERVENTIONS: Both groups received 12 hours of the Arthritis Self Management program over 12 weeks and were followed for an additional 12 weeks. In addition, the Walk + group received individualized instruction in the use of a pedometer, with the goal of increasing their step count by 30% of their baseline step count. MEASUREMENTS: The outcome measures were physical activity (daily step counts and total activity vector magnitude as measured by a pedometer and Tritrac R3D accelerometer), quadriceps femoris strength (isometric peak torque), and functional performance tasks (100-foot walk-turn-walk, timed stair climb, timed chair rise, and pain status). RESULTS: Daily steps walked showed a significant group-by-time interaction (P =.04) after controlling for age. From baseline to completion of training, a 23% increase in daily steps occurred in the Walk + group and a 15% decrease in the EDU group. Although steps increased in the Walk + group, total activity vector magnitude was maintained, suggesting a more efficient gait. The Walk + group became quicker than the EDU group in the normal pace walk-turn-walk (P =.04). An isometric strength gain of 21% postintervention was seen in the Walk + group, compared with a loss of 3.5% in the EDU group. CONCLUSION: In older adults with symptomatic knee OA, Walk + appears to increase walking, with improvements in muscle strength and walking performance. The use of a home-based pedometer-driven program to increase physical activity, strength, and function in this population warrants further research. PMID- 12588584 TI - Environmental components of mobility disability in community-living older persons. AB - OBJECTIVES: To examine the relationship between characteristics of the physical environment and mobility disability in community-living older persons. DESIGN: Cross-sectional study conducted on three groups of community-dwelling older adults. SETTING: Community-dwelling older people in Seattle, Washington, and Waterloo, Ontario, Canada. PARTICIPANTS: Fifty-four older adults (> or =70) were recruited from two geographic sites and grouped according to level of physical function (elite, physically able, physically disabled). MEASUREMENT: Subjects reported on frequency of encounter versus avoidance of 24 features of the physical environment, grouped into eight dimensions, using a five-point ordinal scale (never, rarely, sometimes, often, always). Never and rarely responses were combined and coded as not encountered or not avoided, whereas the sometimes, often or always responses were combined and coded as encountered or avoided. RESULTS: Disabled older adults reported fewer encounters with and concomitantly greater avoidance of physical challenges to mobility than nondisabled older adults. However, both encounter and avoidance varied by environmental dimension. CONCLUSION: Results support the hypothesis that mobility disability results from an interaction of individual and environmental factors. Mobility disability is associated with avoidance of some, but not all, physically challenging features within the environment, suggesting that some environmental features may disable community mobility more than others. PMID- 12588585 TI - Hospital readmissions after hospital discharge for hip fracture: surgical and nonsurgical causes and effect on outcomes. AB - OBJECTIVES: To examine the causes of hospital readmission after hip fracture and the relationships between hospital readmission and 6-month physical function and mortality. DESIGN: Prospective, multisite, observational cohort study. SETTING: Four hospitals in the New York City metropolitan area. PARTICIPANTS: Five hundred sixty-two patients hospitalized for hip fracture aged 50 and older and discharged alive in 1997-1998. MEASUREMENTS: Patient demographic characteristics, type of fracture and repair, comorbid conditions, postoperative complications, do not resuscitate status, and active clinical problems at the time of hospital discharge. Prefracture and 6-month mobility were measured using the Functional Independence Measure. Hospital readmissions and International Classification of Diseases, Ninth Revision principal diagnoses were ascertained from hospital admission/discharge databases, the New York Statewide Planning and Research Cooperative System, medical record review, and patient self-report. RESULTS: Eighty-two percent of participants were women, and 93% were white. Within 6 months after hospital discharge, 178 (32%) patients were readmitted to the hospital, with 45 (8%) readmitted more than once. Forty-seven of 233 readmissions (20%) occurred within the first 2 weeks after discharge, and 80 (34%) occurred within 4 weeks. Over 6 months, 89% of readmissions were for nonsurgical problems, of which infectious (21%) and cardiac (12%) diseases were the most common. In multivariate analyses, patients who were readmitted were more likely to require total assistance with ambulation at 6 months (odds ratio (OR) = 2.7, 95% confidence interval (CI) = 1.6-4.6) and to die (OR = 4.0, 95% CI = 2.2-7.3) than those not readmitted. CONCLUSION: Hospital readmissions after hip fracture are largely due to nonsurgical illness and are associated with increased morbidity and mortality. PMID- 12588586 TI - Sociodemographic and health characteristics of older Chinese on admission to a nursing home: a cross-racial/ethnic study. AB - OBJECTIVES: To investigate sociodemographic characteristics (SDCs) and health status of older Chinese newly admitted to a nursing home (NH) and to compare them with the characteristics of residents from other racial/ethnic groups. DESIGN: Secondary analysis of the admission Minimum Data Set Plus (MDS+). SETTING: A New York City municipal NH near Chinatown. PARTICIPANTS: Two hundred fifty-eight (125 Chinese, 57 white, 53 Hispanic, and 23 black) of 292 residents consecutively admitted from November 1992 to May 1997 were selected after excluding those younger than 60 or transferred from another NH. MEASUREMENTS: SDCs, health status parameters (cognitive performance, physical functioning, mood/behavior patterns, and psychosocial well-being), and morbidity information (most-frequent diagnoses/conditions and medication use) documented in or generated from the MDS+. RESULTS: The majority of these Chinese were first-generation immigrants and spoke primarily Cantonese or Mandarin Chinese. Compared with whites, they were more likely to be married, less likely to have lived alone, more likely to be using Medicaid, less likely to make medical decision alone, and more likely to depend on family members for decision-making. Nearly three-quarters of Chinese had cognitive impairment. There was an underdiagnosis of dementia in the Chinese subjects on admission. Severe dependence in activity of daily living was identified in more than one-third of Chinese. Fewer Chinese were using psychotropic medications on admission than the whites. Similar to other groups, many of the Chinese subjects were incontinent of bowel and bladder and had chewing or swallowing problems, hypertension, anemia, and stroke. CONCLUSION: This is the first systematic report of the SDCs and health status of a group of newly admitted older Chinese to an urban NH in the United States using the Minimum Data Set database. These findings suggest that Chinese residents are as frail as other racial/ethnic residents on admission. NHs caring for older Chinese need to be sensitive to the presence of dementia, and require a staff that can speak Cantonese and Mandarin Chinese and is comfortable negotiating with families who are more likely to be the designated decision makers. PMID- 12588587 TI - Association between dementia and midlife risk factors: the Radiation Effects Research Foundation Adult Health Study. AB - OBJECTIVES: To investigate the association between midlife risk factors and the development of vascular dementia (VaD) or Alzheimer's disease (AD) 25 to 30 years later. DESIGN: A prevalence study within a longitudinal cohort study. SETTING: Subjects in the Adult Health Study (a prospective cohort study begun in 1958) have been followed through biennial medical examinations in Hiroshima, Japan. PARTICIPANTS: One thousand seven hundred seventy-four subjects in Hiroshima, Japan born before September 1932 (1,660 with no dementia, 114 with dementia (51 with AD, and 38 with VaD) diagnosed from 1992 to 1997 according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria). MEASUREMENTS: The subjects were examined for effect on dementia of sex, age, education, atomic bomb radiation dose, and midlife factors associated with risk (smoking, alcohol intake, physical activity, dietary habits, systolic blood pressure (SBP), body mass index, and history of diabetes mellitus) that had been evaluated in 1965 1970. RESULTS: VaD prevalence increased significantly with age, higher SBP, and lower milk intake. The odds ratios of VaD for age (in 5-year increments), SBP (10 mmHg increments), and milk intake (almost daily/less than four times a week) were 1.29, 1.33, and 0.35, respectively. The risk factors for VaD were compatible with the risk factors for stroke in this study population. AD prevalence increased significantly with age and lower education. Other midlife factors and radiation dose did not show any significant association with VaD or AD. CONCLUSION: Increased SBP and low milk intake in midlife were associated with VaD detected 25 to 30 years later. Early behavioral control of the risk factors for vascular disease might reduce the risk of dementia. PMID- 12588588 TI - Cognitive screening using a tape recorder: a pilot study. AB - OBJECTIVES: To determine whether a tape recorder can be used to administer cognitive tests efficiently and yield valid results. DESIGN: Convenience sample. Administration of cognitive test materials by tape recorder and conventional technique. SETTING: Outpatient clinic. PARTICIPANTS: Subjects from memory disorder clinic, hostel accommodation, and community. MEASUREMENTS: Responses to Hopkins Verbal Learning Test-revised, verbal fluency items from the controlled oral word association test, 10-item naming task, a construction task, and speed writing task. RESULTS: Performances on the tape- and clinician-administered battery of tests were highly correlated. Memory impairment was accurately detected using the tape battery. Data from 30 minutes of testing via tape were obtained at the cost to the clinician of 2 to 3 minutes of scoring time. CONCLUSION: Tape-administration of cognitive test material warrants further study as an efficient means of cognitive screening. PMID- 12588589 TI - An evaluation of a low-intensity intervention to introduce medical students to healthy older people. AB - This is a report of a controlled, prospective, longitudinal trial of an intervention to affect medical students' attitudes toward aging. Members of the Class of 2002 at the University of Oklahoma College of Medicine were assigned a senior mentor (a community-dwelling older person) upon matriculation into medical school. Students were required to perform a structured interview with the mentor once per semester for the first 2 preclinical years and to discuss these interviews in small groups mediated by geriatrics faculty. Members of the Class of 2001 were controls. Attitudes toward aging were determined using the Aging Semantic Differential (ASD) attitude scale in August 1998 and again at the end of the second year of medical school. Initial mean ASD scores were not significantly different for the two groups. Although both classes experienced improvements in their ASD scores from Time 0 to Time 1, the improvement for the class of 2002 was significantly greater than that for the class of 2001 (2001 class mean = 0.17, 2002 class mean = 0.40, t = -3.09, degrees of freedom (df) = 219, P =.002). This difference held up under controls for sex, age, prior visits to a nursing home, prior work/volunteering in an old-age environment, and a prior course on aging (Model F = 3.00, df = 6/214, P =.008; class F = 9.70, df = 1, P =.002). It was concluded that a low-intensity intervention to introduce entering medical students to healthy older people might have a positive effect on attitudes toward aging. PMID- 12588590 TI - Effect of terazosin on clinical benign prostatic hyperplasia in older adults. AB - PURPOSE: The objective of this meta-analysis of randomized controlled trials (RCTs) of terazosin versus placebo in men with benign prostatic hyperplasia (BPH) was to evaluate the effect of terazosin on lower urinary tract symptoms. BACKGROUND: BPH is common in older men; more than 90% of men aged 70 and older suffer from BPH. (1,2) BPH has substantial negative effect on quality of life, with symptoms that interfere with activities of daily living. (3) The symptoms of BPH are primarily due to bladder outlet obstruction and are caused by dynamic prostatic smooth muscle tone and static prostatic obstruction. (4,5) The smooth muscle tone is dependent on the degree of the alpha-1 adrenergic receptors present in the bladder neck, prostrate capsule, and prostrate gland. The blockade of alpha-1 receptors with drugs such as terazosin reduces bladder outlet obstruction by reducing the smooth muscle tone. STUDY SELECTION AND DATA SOURCES: Selected studies were all RCTs of terazosin with two primary end points, namely, changes in the peak urinary flow rates and urinary symptom scores from baseline. Nine studies were selected using a Medline search, the details of which were not provided. The studies were conducted in North America and Europe between 1992 and 1996. Two of the studies included unpublished data that were obtained from Abbott Laboratories, which is the manufacturer of terazosin (Hytrin). The company also provided data on the other seven studies. DATA EXTRACTION: The meta-analysis did not provide details of the data abstraction process. Urinary symptom score was measured using two different tools: the Boyarsky Symptom Index (BSI) and American Urology Association Symptom Index (AUA-SI). The BSI was used in seven studies and AUA-SI in two studies. The authors calculated a common symptom score, based on the common items from both questionnaires, that had all but one item used in the AUA-SI. Questions from AUA-SI are scored on a 0 to 5 scale and those on BSI on a 0 to 3 scale. To adjust for this difference, the authors performed a nonlinear rescaling of the AUA-SI to adjust for these scoring differences. MAIN RESULTS: The ages of the patients (N = 3,948) ranged from 62 to 66. The duration of the studies varied from 8 to 52 weeks. Four of the studies used fixed dosages, and the rest used titration-to-response doses. The effect of terazosin versus placebo on peak urinary flow rate was similar in all the studies. The estimated mean increase in peak urinary flow rate was 1.39 mL/s greater in men receiving terazosin than in those receiving placebo, and the improvements were observed within 8 weeks of therapy. However, the changes in the symptom scores varied across the studies. Patients treated with terazosin had an estimated 1.9-point reduction (95% confidence interval (CI) = 1.6-2.3) in BSI and 3.5-point reduction (95% confidence interval (CI) = 2.8-4.1) in AUA-SI score from baseline. Improvements in scores were greater in studies that were conducted for longer duration. CONCLUSIONS: The authors concluded that treatment with terazosin was associated with greater peak urinary flow rate and lower symptom score than treatment with placebo. There was no variability of efficacy of terazosin as a result of prostrate volume, and measurement of prostrate volume was not essential before initiation of therapy. PMID- 12588591 TI - Vitamin D and quadriceps exercise--got milk? Got a chair? PMID- 12588592 TI - Enhanced ambulation and quality of life in advanced Alzheimer's disease. PMID- 12588593 TI - Preparation for healthy retirement. PMID- 12588594 TI - Alterations in cognitive function during the menopausal transition. PMID- 12588595 TI - Periventricular white matter lesions and sleep alteration in older people. PMID- 12588596 TI - Azathioprine and low-dose corticosteroids for the treatment of cryptogenic organizing pneumonia in an older patient. PMID- 12588599 TI - Incidence and growth pattern of simple cysts of the kidney in patients with asymptomatic microscopic hematuria. AB - BACKGROUND: We examined the incidence and natural history of simple renal cysts found by ultrasonography (US) in patients referred for asymptomatic microscopic hematuria. METHODS: Among the 906 patients aged 18-78 years, 743 patients who had undergone US were included in the present study. The natural history of simple renal cysts was investigated in 55 patients who underwent periodical US examinations for more than 3 years. RESULTS: The incidence of simple renal cysts was 4.3% for ages 29 years or younger, 15.3% for ages 30-39, 21.8% for ages 40 49, 23.3% for ages 50-59 and 32.6% for ages 60 years or older; thus the incidence increased in older age groups (P = 0.0005 for men, P = 0.0020 for women). Men tended to have a higher incidence than women. The degree of hematuria did not influence the incidence of renal cysts (P = 0.9044). The annual growth rate of the mean maximum diameter was 4.2% during a 3-year follow-up period in 55 patients and 5.1% during a 6-year follow-up in 31 patients. CONCLUSION: Since the diameter of a renal cyst may increase by 5% annually, the diameter of the cyst may increase by 1.6 times in 10 years. The scheduling of follow-up examinations depends on the size at the time of disclosure, the effects on calyceal systems, or the suspicion of a concurrent malignant disease. However, the most simple renal cysts may be followed-up at an interval of more than 10 years, once a diagnosis has been established. PMID- 12588600 TI - Predictors of prostate cancer on repeat transrectal ultrasound-guided systematic prostate biopsy. AB - BACKGROUND: We analyzed the outcome of repeated transrectal ultrasound (TRUS) guided systematic prostate biopsy in Japanese men whose clinical findings were suspected of prostate cancer after previous negative biopsies. METHODS: Between January 1993 and March 2002, 1045 patients underwent TRUS-guided prostate biopsy. Among them, 104 patients underwent repeat biopsy due to indications of persistent elevated serum prostate-specific antigen (PSA), abnormal digital rectal examination (DRE) or TRUS, increased PSA velocity, and/or previous suspicious biopsy findings. Several clinicopathological factors were evaluated for their ability to predict the detection of prostate cancer on repeat biopsy. RESULTS: Prostate cancer was detected in 22 of 104 patients (21.2%) who underwent repeat biopsies. PSA concentration and PSA density at both the initial and repeat biopsies, and PSA velocity in men with positive repeat biopsy were significantly greater than those in men with negative repeat biopsy. The incidence of abnormal findings in DRE and TRUS at initial biopsy in men with positive repeat biopsy was also significantly higher than that in men with negative repeat biopsy. However, neither the presence of prostatic intraepithelial neoplasia nor number of biopsy cores at initial biopsy had a significant association with the results of the repeat biopsy. Furthermore, multivariate analysis revealed that PSA and PSA density at both the initial and repeat biopsies, PSA velocity, and DRE and TRUS findings at initial biopsy were independent predictors of malignant disease on repeat biopsy. CONCLUSION: Despite an initial negative biopsy, repeat TRUS-guided biopsy should be carried out to exclude prostate cancer in cases of suspicious clinical findings, such as elevated PSA or PSA-related parameters, or abnormal findings of DRE or TRUS. PMID- 12588601 TI - Protecting spermatogonia from apoptosis induced by doxorubicin using the luteinizing hormone-releasing hormone analog leuprorelin. AB - PURPOSE: The present study was performed to investigate the protective effect of leuprorelin (LH-RH analog), on spermatogonia apoptosis induced by doxorubicin (DXR) in the Sprague-Dawley rat model. METHODS: Twenty-four adult male rats were divided into the following four groups: (i) control group; (ii) group given doxorubicin (intravenous injection, 8 mg/kg); (iii) group given leuprorelin (subcutaneous injection, 3 mg/kg); and (iv) group given both doxorubicin (intravenous injection, 8 mg/kg) and leuprorelin (subcutaneous injection, 3 mg/kg). Evaluation for quantification of apoptotic spermatogonia was made by the ratio of TUNEL-labeled spermatogonia versus 100 Sertoli cells in each seminiferous tubule. Two hundred seminiferous tubules of each rat were assessed. RESULTS: The ratio of apoptotic spermatogonia versus 100 Sertoli cells at stages II-IV of the groups given DXR (groups 2 and 4) were significantly higher than those of the other groups. However, the value at stages II-IV of the group given both DXR and leuprorelin (group 4) was significantly lower than that of the group given DXR (group 2). CONCLUSION: The significant prophylactic effect (P < 0.05) of LH-RH analog against doxorubicin-induced spermatogonial apoptosis was observed in a stage specific manner by microscopic evaluation with TUNEL. PMID- 12588603 TI - Concentration gradient of oxalate from cortex to papilla in rat kidney. AB - BACKGROUND: The kidney eliminates the major fraction of plasma oxalate. It is well known that oxalate is freely filtered by glomeruli and secreted by the proximal tubules. However, the renal handling of oxalate in distal nephrons, which is considered as playing an important role in stone formation, remains obscure. METHODS: At 15-180 min after intravenous injection of 14C-oxalate to rats, the intrarenal localization of radioactivity was quantitatively measured by the radioluminographic method using a bioimaging analyzer. Tissue radioactivity was compared with plasma, and urinary radioactivities were measured by a liquid scintillation counter. The control study was conducted with 14C-inulin. RESULTS: The radioactivity of 14C-oxalate in the papilla was 10 times greater than in the cortex and eight times greater than in the medulla 180 min after injection when almost no radioactivity was present in the urine. In contrast, the radioactivity of 14C-inulin was nine times less in the papilla than in the cortex at the same time. CONCLUSION: Oxalate remains in the renal papilla for an extended period. This accumulation of oxalate may be attributed to calcium oxalate crystal fixation along the deep nephron which is considered to be the first step of stone formation. PMID- 12588602 TI - Inhibitory effect of clotrimazole on angiogenesis associated with bladder epithelium proliferation in rats. AB - BACKGROUND: Up-regulation of epithelial vascular endothelial growth factor (VEGF) expression was recently reported at an early stage of bladder carcinogenesis in rats. Subsequently, clotrimazole (CLT) was reported to exert an inhibitory effect on the in vitro proliferation of vascular endothelial cells stimulated by VEGF. The present study was performed to evaluate the effect of CLT on the initial stage of N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN)-induced carcinogenesis of the urinary bladder in rats. METHODS: Seven-week-old-female Wistar rats were divided into both a control group and the other four groups that were allowed free access to drinking water containing 0.05%BBN. Two groups of rats (10-CLT and 20-CLT) received CLT (100 mg/kg per day) once daily by gavage for 10 and 20 weeks, respectively. The other two groups of rats (10-BBN and 20-BBN) were maintained on the same BBN-containing drinking water for 10 and 20 weeks, respectively, without receiving CLT. After removing the urinary bladder of each rat, expression of VEGF and VEGF receptor FLT1, and new capillaries were examined by immunohistochemical staining. VEGF mRNA was detected by the reverse transcription polymerase chain reaction. RESULTS: The ratio of VEGF-positive cells and the expression of VEGF mRNA were lower in the 20-CLT group than in the 10-BBN group. Newly formed capillaries in the 20-CLT group were significantly decreased compared with those in the 10-BBN group. CONCLUSION: These quantitative data suggested that CLT can inhibit VEGF expression in vivo and hence inhibit angiogenesis in the early stages of bladder carcinoma. PMID- 12588604 TI - Laparoscopic correction of vesicoureteral reflux using the Lich-Gregoir technique: initial experience and technical aspects. AB - We report two female cases with vesicoureteral reflux (VUR) who underwent laparoscopic correction of VUR by the Lich-Gregoir technique. One patient was 10 years of age, with international grade III reflux of the left ureter; the other was 27 years of age, with grade II reflux of the right ureter. The respective operating times were 6 h 40 min and 6 h 10 min. There were no complications during surgery. Although post-surgical ultrasonography revealed mild hydronephrosis in the adult patient, the condition resolved spontaneously 2 months after surgery. The disappearance of reflux was confirmed in both cases by voiding cystography 6 months after surgery. PMID- 12588605 TI - Extraperitoneal laparoscopic Lich-Gregoir antireflux plasty for primary vesicoureteral reflux. AB - We performed laparoscopic Lich-Gregoir antireflux plasty on 4 patients with primary vesicoureteral reflux. All procedures were conducted using the extraperitoneal approach. The average surgical time was 230 min. There were no complications. After surgery, voiding cysturethrograms showed no reflux in all patients. PMID- 12588607 TI - Chromophobe cell renal carcinoma with acquired cystic disease of the kidney in a long-term hemodialysis patient. AB - We report a rare case of chromophobe cell renal carcinoma found in a 52-year-old female who had received hemodialysis therapy for 13 years. She was diagnosed as having a left renal tumor 7.5 cm in diameter with acquired cystic disease of the kidney (ACDK) by ultrasonographic examination during periodical systemic screening. As abdominal computed tomography scanning and enhanced color Doppler ultrasonography suspected that the hypervascular tumor was renal cell carcinoma, she underwent translumbar nephrectomy in July 2000. The histopathological diagnosis was chromophobe cell carcinoma with pT2 and grade 2 malignancy. Chromophobe cell carcinoma is uncommon among renal tumors with ACDK found in long term hemodialysis patients. PMID- 12588608 TI - Extensive cutaneous metastasis of transitional cell carcinoma of the bladder. AB - Cutaneous metastasis of bladder carcinoma is extremely rare. Iatrogenic implantations have been the main cause in the majority of cases of transitional cell carcinoma with cutaneous metastasis. Otherwise, primary cutaneous metastasis is accepted as the late manifestation of systemic spread. The present paper describes a case of relatively early and extensive skin metastasis of transitional cell carcinoma in a 78-year-old man. The patient had a histopathological diagnosis of poorly differentiated (grade III) muscle invasive transitional cell carcinoma with a staging of T2NOMO 6 months prior to presenting. He presented to our outpatient clinic with a 3-month history of skin lesions as multiple, rubbery subcutaneous nodules. Radiological reinvestigation revealed no other metastatic site (including bone and lung), except for a metastatic nodule in the liver. The present paper reports an interesting and rare case of extensive skin metastasis of transitional cell carcinoma as the primary complaint. PMID- 12588609 TI - Advanced ureteral cancer with complete remission achieved by taxan containing systemic chemotherapy. AB - We report a case of advanced ureteral cancer successfully treated with systemic chemotherapy combined with irradiation. A 47-year-old man was diagnosed as having a right ureteral cancer at the clinical stage of T4, N2 and M1 (liver). A papillary tumor was also found in the bladder and the resected specimen showed a grade 1 transitional cell carcinoma. Although three cycles of methotrexate, vinblastine, pirarubicin and cisplatin (MVAC) gave partial response to the ureteral tumor, new metastases to the lung and pelvic bone were observed. The patient received 50 Gy external irradiation to the pelvis, 11 cycles of paclitaxel (270 mg) and cisplatin (60-80 mg) followed by four cycles of docetaxel (100 mg) and cisplatin. Thereafter, he underwent bone biopsy, partial hepatectomy, total nephroureterectomy and lymph node resection, by which a complete response was achieved pathologically. The patient has been alive without evidence of disease for 12 months. PMID- 12588610 TI - Prostatic carcinosarcoma: a case report and review of literature. AB - A unique case of carcinosarcoma of the prostate detected in a 71-year-old man is presented. Pelvic exenteration was performed, and the resected prostatic mass was found to consist of two histologically distinct elements; adenocarcinoma and sarcoma with focal osteosarcomatous element. The patient is still alive with neither metastasis nor recurrence. This is the 42nd case of carcinosarcoma of the prostate to be reported in the literature. PMID- 12588611 TI - Squamous cell carcinoma of the prostate. AB - Squamous cell carcinoma of the prostate is rare, accounting for 0.5-1% of all prostatic cancers. It is highly aggressive and responds poorly to any mode of therapy. We present a case of squamous cell carcinoma of the prostate that developed in a patient with prostatic adenocarcinoma following radiation therapy. PMID- 12588612 TI - Telomerase activity in urine after transurethral resection is not a predictive marker for recurrence of superficial bladder cancer. PMID- 12588614 TI - Focus group study of endometriosis: struggle, loss and the medical merry-go round. AB - Women with endometriosis experience a range of problems for which they may or may not be adequately supported. This paper reports on one aspect of a study conducted at the Epworth Hospital, Melbourne, to identify the information needs of women facing laparoscopy for endometriosis. A number of focus groups were conducted that provided women with a forum for communicating their experiences of endometriosis and laparoscopy. The findings include the experiences of 61 women who described the lack of support, the struggles and the losses involved in living with endometriosis. By far the worst experience that these women described was the encounter with health professionals and the ways in which their symptoms were trivialised and dismissed. There is a great deal for nurses to learn about the experience of living with endometriosis if they are to support women with this chronic illness in their search for well-being. PMID- 12588615 TI - Women's experiences of Caesarean section and vaginal birth after Caesarian: a Birthrites initiative. AB - This paper reports the findings of a small pilot study undertaken to identify women's perceptions of their Caesarean section and/or their experience of attempting to achieve a vaginal birth after Caesarean. Fifty-nine women replied to a survey designed and distributed by the consumer organisation Birthrites. The survey consisted of 22 closed and open-ended questions. Descriptive statistics and content analysis were used to analyse the data. Six major factors that impacted on women's perceptions of their birthing experience were identified: (i) 'being supported'; (ii) 'violated expectations'; (iii) 'loss of control'; (iv) 'health professionals' language, attitudes and care practices'; (v) 'the labour experience and the cascade of intervention'; and (vi) 'surgical birth and the separation from the baby'. The results support previous work in this area and demonstrate that some women suffer from psychological trauma following an emergency Caesarean section. The experiences of the women in this small study indicate a need for health professionals to not only better understand the distress that may occur but to also develop a greater understanding of the impact of their interactions with women. The study also provides a consumer 'voice' on current routine hospital policies surrounding vaginal birth after Caesarean and confirms the importance of choice, control and continuity of care for all women regardless of their 'risk' status. PMID- 12588616 TI - Quality of life of elderly Japanese patients with glaucoma. AB - To evaluate the quality of life (QOL) of elderly Japanese glaucoma patients, we performed a survey of 114 outpatients of a university eye clinic aged from 47 to 86 years. We examined 110 cases using one-way anova, Bonferroni's multiple comparison, non-parametric Mann-Whitney U-test and Pearson's correlation coefficient analysis. The group with better traditional visual measures had better QOL than the group with worse visual acuity. In the elderly groups of patients, the loss of hopes for future life and its consequences caused greater reduction in QOL than did the symptoms of glaucoma. The reliability of our questionnaire was high as determined using alpha-coefficients of Cronbach reliability analysis. Our questionnaire was reliable and useful for the assessment of QOL of elderly Japanese glaucoma patients. PMID- 12588617 TI - Nurses' influence on quality of care in postoperative pain management: a phenomenological study. AB - The aims of this study were to describe a group of nurses' influence on the quality of care in postoperative pain management and to elucidate the meaning of exercising this influence. Fifteen protocols, written by experienced clinical nurses, describing complex postoperative pain situations where the nurses' actions influenced the outcome were analysed by using two different phenomenological methods. The results showed that the general meaning structure of the nurses' influence on the quality of care in postoperative pain management consisted of (i) the nurses' perception of an unsatisfactory situation concerning the pain management of the surgical patients; (ii) that the nurses personally intervened; and (iii) that they changed the outcome of the situation in a positive direction. The meaning of exercising this influence was interpreted as an aspiration to relieve the patients from their suffering by exercising professional skills and knowledge in a creative, problem-solving, caring process. PMID- 12588618 TI - Development of a group model of clinical supervision to meet the needs of a community mental health nursing team. AB - Clinical supervision is an important tool in the development of quality nursing care. It involves a process of reflection upon practice, the aim of which is to improve clinical practices and hence improve patient outcomes. The term 'clinical supervision' is itself problematic in that it implies an hierarchical, rather than a nurse-centred and reflective, process. In addition there are a variety of models of supervision which range from the purely managerial to the clinical. This gives rise to confusion and in some cases suspicion, in clinicians. This paper reports on the development, implementation and evaluation of a group model of clinical supervision developed by a small team of mental health nurses in a community mental health setting. This team recognised the need for a formal clinical supervision model but was unsure as to the model which best suited their practice situation and needs. Through collaboration with a university department of nursing, this group developed its own model of group clinical supervision. This paper reports on the development of the model and its evaluation. The model was developed with a small team of community nurses and hence may not be applicable to other teams and other settings. However, the methods described may be useful as a guide to other nurses who wish to plan, implement and evaluate a model of clinical supervision in their workplace. PMID- 12588619 TI - Evaluation of quality of life in Chinese patients with gynaecological cancer: assessments by patients and nurses. AB - Quality of life (QOL) has recently become one of the most important focuses in oncology nursing practice and research. The aims of this study were to evaluate the QOL in Chinese cancer patients under chemotherapy and to explore the discrepancy between patients' and nurses' assessments of the patients' conditions. The study participants included gestational trophoblastic disease patients (n = 68), ovarian cancer patients (n = 105), patients with other gynaecological cancer (n = 18), and their attending nurses (n = 25) at hospitals affiliated with universities in Beijing. The high level of some symptom subscales/items in patients suggested that innovative and more effective interventions should be developed and utilised in nursing practice. Patients with metastasis reported lower levels of QOL than those without metastasis. The nurses in this study tended to estimate the patients' QOL inaccurately, except for a few objective aspects. The results of this study can help nurses in planning interventions to enhance the QOL of patients with gynaecological cancer undergoing chemotherapy. The nurses need to be trained to become more sensitive and competent in assessing patients' subjective information. PMID- 12588620 TI - Effects of sleep loss in men and women with insufficient sleep suffering from chronic disease: a model for supportive nursing care. AB - This study evaluated self-reported subjective health and effects of sleep loss according to perceived interfering cognitive anxiety related to falling asleep in patients with and without insufficient sleep and gender differences in these aspects 5 years after coronary artery bypass graft and transluminal coronary angioplasty. A total of 145 patients, five years after intervention, responded to a mailed questionnaire. Nearly 60% had severe combined sleep disturbances; 35.9% of these had complained of insufficient sleep and 15% also perceived difficulty falling asleep related to cognitive anxiety. Measurable gender differences were small. A theoretical framework is presented which can increase understanding among nurses, patients and their relatives concerning the quality and quantity of sleep and sleep loss related to quality of life. These results suggest that there are significant relationships between sleep quality, resilience to stress and coping strategy in patients with a chronic disease, indicating the need for more individualised supportive nursing care. PMID- 12588621 TI - Pulmonary thromboendarterectomy in the UK. AB - This paper describes the disease of chronic thromboembolic pulmonary hypertension. This is a disease with an extremely poor prognosis, and the surgical procedure of pulmonary thromboendarterectomy represents the only possibility of a cure for patients with this condition. Further the paper describes the role and responsibilities of critical care nurses, specialist nurses in palliative care and pulmonary hypertension when caring for these patients before and after surgery. PMID- 12588622 TI - Nursing management of fever in children: a systematic review. AB - OBJECTIVES: The aim of the present review was to determine whether the best available evidence supports the types and timing of the various nursing interventions that are commonly used to reduce fever in non-critically-ill children, and to what extent the outcomes are influenced by these nursing actions. METHODS: Studies included were randomised or quasi-randomised controlled trials that involved non-critically-ill children with a fever aged between 3 months and 16 years. The search strategy sought to identify both published and unpublished research reports in the English language and covered all major databases up to 1998. The methodological quality of each study was assessed by two independent reviewers using a piloted critical appraisal checklist. Despite all studies being randomised, heterogeneity precluded conduction of a meta analysis; therefore, evidence was synthesised using narrative summaries. RESULTS: Ten studies were assessed as being of sufficient quality to be included in the review. These studies addressed two of the intervention categories identified in the protocol: (i) administration of antipyretics (paracetamol); and (ii) direct cooling measures on the outcome measure (reduction of or prevention of increase in fever). The review found little benefit from sponging in temperate climates and usually at the expense of the child's comfort. There may be situations in high environmental temperatures and high humidity, or where there is a need for immediate temperature reduction, in which sponging may be warranted. Risks were identified when paracetamol was administered on a sustained basis over even a short period of time and above a relatively low total daily dosage. There was a lack of evidence to support the administration of antipyretics to reduce the incidence of febrile convulsions. There is a need for parental education that focuses on knowledge of the body's protective physiological responses and how to support these responses. CONCLUSION: The primary purpose for intervening when a child has a fever is to increase the child's comfort. This consideration should be weighed against any harm that might result from intervening. There was a lack of evidence to support the routine use of sponging. The administration of paracetamol should be used selectively and with caution. In summary, care needs to be individualised, based on current knowledge of the effectiveness and risks of interventions. PMID- 12588625 TI - Nitric oxide: therapeutic opportunities. AB - Fifteen years after the discovery of nitric oxide as a biological mediator how close are new therapies? This article describes the roles of nitric oxide, illustrates how its discovery is altering the way in which certain established drugs are being used and reviews new therapeutic developments. PMID- 12588626 TI - The therapeutic potential of NO-NSAIDs. AB - NSAIDs, including those that are selective for cyclooxygenase-2, are among the most widely used drugs. However, these drugs produce significant side effects in the gastrointestinal and cardiorenal systems, which greatly limit their utility. In recent years, a new type of anti-inflammatory agent has been developed that appears to offer significant advantages over conventional and Cox-2-selective NSAIDs. No-NSAIDs are derivatives of conventional NSAIDs, which are able to release nitric oxide over prolonged periods of time. The combination of balanced inhibition of the two main isoforms of COX with controlled release of nitric oxide yields a series of drugs that exert anti-inflammatory and analgesic activities in a wide range of settings, and have markedly reduced gastrointestinal and cardiorenal toxicity. Recent clinical trials of NO-NSAIDs have provided a 'proof of concept' that is completely consistent with pre clinical characterization of these compounds. PMID- 12588627 TI - Internationalization of regulatory requirements. AB - The aim of harmonisation of medicines regulatory requirements is to allow the patient quicker access to new drugs and to avoid animal and human duplications. Harmonisation in the European Union (EU) is now completed, and has led to the submission of one dossier in one language study leading to European marketing authorizations, thanks in particular to efficacy guidelines published at the European level. With the benefit of the European experience since 1989, more than 40 guidelines have been harmonised amongst the EU, Japan and the USA through the International Conference on Harmonisation (ICH). ICH is a unique process gathering regulators and industry experts from the three regions. Its activity is built on expertise and trust. The Common Technical Document (CTD), an agreed common format for application in the three regions, is a logical follow-up to the ICH first phase harmonising the content of the dossier. The CTD final implementation in July 2003 will have considerable influence on the review process and on the exchange of information in the three regions. PMID- 12588628 TI - Pharmacogenetics of the major polymorphic metabolizing enzymes. AB - There is increasing information available on the existence of polymorphisms in genes encoding xenobiotic metabolizing enzymes and the functional significance of many of these. In addition to genes long recognized as being polymorphic, such as CYP2D6, CYP2C19 and CYP2C9, there is now information available on the existence of polymorphisms in other cytochrome P450 genes such as CYP2A6, CYP2B6 and CYP2C8. With respect to phase II metabolism, polymorphisms in GSTM1, GSTT1, NAT2 and TPMT are well understood but information is also emerging on other GST polymorphisms and on polymorphisms in the UDP-glucuronosyltransferases and sulfotransferases. The availability of comprehensive information on the occurrence and functional significance of polymorphisms affecting drug metabolism should facilitate their application to pharmacogenomic profiling. PMID- 12588629 TI - Impairment of vascular and platelet levels of nitric oxide and cyclic guanosine 3',5'-monophosphate in cyclosporin A-induced hypertensive rats. AB - The therapeutic use of the immunosuppressive agent cyclosporin A (CsA) is associated with arterial hypertension and increased risk of thromboembolism. Impaired endothelium-mediated relaxation is one of the main hypotheses explaining the CsA-induced vascular hyper-reactivity. Since nitric oxide (NO) modulates both vascular and platelet activity, we studied the effects of CsA on the levels of arterial and platelet NO as well as 3',5'-cyclic guanosine monophosphate (cGMP) levels which are influenced by NO. An animal model of CsA-induced hypertension was used. Wistar rats were treated with a clinically relevant, oral dose of 5 mg/kg CsA, daily, for 4 weeks. CsA increased both systolic and diastolic blood pressures compared to non-treated rats (P < 0.01). Nitrite, a NO metabolite, decreased in the entire aorta wall (30%, P < 0.05) and in the aorta wall without the endothelial layer (70 %, P < 0.05) in CsA-treated vs. control groups. cGMP content was also decreased in the CsA-treated group (67%, P < 0.01) vs. control. Taken together, these results suggest a defect on the endothelial NO generation, acceleration of breakdown and/or consumption of NO, as well as marked alterations directly on cGMP metabolism. Conversely, platelet nitrite and cGMP content significantly increased in the CsA-treated rats, which was also observed in in vitro studies of platelet nitrite release following CsA treatment. This suggests a platelet self-regulation mechanism against CsA-induced platelet hyper reactivity, which, in turn, could compensate vascular impairment. PMID- 12588630 TI - Trimetazidine effect on phospholipid synthesis in ventricular myocytes: consequences in alpha-adrenergic signaling. AB - The anti-anginal drug trimetazidine (TMZ) has been shown to increase the synthesis of phospholipids in ventricular myocytes, including phosphatidyl inositol (PI). This study focused on the consequences of increasing PI metabolism on alpha-adrenergic signaling pathway in cultured rat cardiomyocytes. In the cells treated with TMZ, the synthesis of PI from inositol was largely increased as compared with the control (+55% in 60 min). The stimulation of alpha adrenergic receptors by phenylephrine (PE) induced a dose-dependent production of inositide phosphates (IPs) by phospholipase C (PLC) activation. However, the amount of available IPs was significantly lower in TMZ-treated cells, in a dose dependent manner. This effect was observed in the presence and absence of the IP1 phosphatase inhibitor LiCl. The in vitro determination of PLC activity revealed that this effect could not be attributed to the direct inhibition of the enzyme by TMZ. The TMZ-induced reduction of IPs in the PE-stimulated cardiomyocytes should be attributed to the increase of inositol recycling and incorporation in membrane structures, elicited by increased phospholipid synthesis. The consequences of this reduction in IPs availability were investigated on the cardiomyocyte hypertrophy induced by alpha-adrenergic chronic stimulation. Acute stimulation with PE increased protein synthesis (+50%), but this increase was largely prevented by TMZ. In conclusion, TMZ reduces cell available IPs, by accelerating their recycling in membranes as PI. This effect results in a cytoprotection in the pathological process of hypertrophy elicited by chronic alpha-adrenergic stimulation. PMID- 12588631 TI - Differential effects of sulphonylureas on the vasodilatory response evoked by K(ATP) channel openers. AB - The potency of three sulphonylureas, glibenclamide, glimepiride and gliclazide in antagonizing the vasorelaxant action of openers of adenosine triphosphate (ATP) regulated K+ channel (KATP) was studied in vivo and in vitro in micro- and macrovessels, respectively. In the hamster cheek pouch, the vasodilatation and the increase in vascular diameter and blood flow induced by diazoxide were markedly reduced by the addition of either glibenclamide or glimepiride (0.8 microm) while they were not affected by gliclazide up to 12 microm. Similarly, in rat and guinea-pig isolated aortic rings, glibenclamide, glimepiride and gliclazide reduced the vasodilator activity of cromakalim. However, the inhibitory effect of gliclazide was considerably less when compared with either glimepiride or glibenclamide. These results suggest that, in contrast to glibenclamide and glimepiride, therapeutically relevant concentrations of gliclazide do not block the vascular effects produced by KATP channel openers in various in vitro and in vivo animal models. PMID- 12588632 TI - Effects of aspirin on N-alpha-tosyl L-arginine methyl ester [TAME]-esterase induced contractions on rat aorta in vitro. AB - The present study was designed to investigate whether N-alpha-tosyl L-arginine methyl ester [TAME]-esterase activation could be the result of endothelial dysfunction. Thoracic aorta from rats was mounted in an organ bath containing Krebs solution. Intact and endothelium denuded aortic strips were challenged with different concentrations of TAME (10(-15)-10(-1) m). The effects of aspirin, a cyclo-oxygenase pathway inhibitor, were also studied on [TAME]-esterase induced contraction on rat aorta strips. Our results showed that aspirin definitely blocked TAME-esterase induced contractions on rat aortic strips. In conclusion, the present work supported the hypothesis that [TAME]-esterase induced contraction in rat aorta in vitro was mediated through release of prostaglandin(s) as a result of endothelial dysfunction. PMID- 12588633 TI - Warming and response to contractile agents in calf cardiac vein: role of the nitric oxide. AB - The effects of warming on the response to various contractile agents of calf cardiac vein were studied using 2.5-mm long cylindrical segments. Concentration response curves for carbachol (10(-9)-3 x 10(-4) m), 5-hydroxytryptamine (5-HT; 10(-8)-3 x 10(-3)), potassium chloride (KCl; 10(-4)-5 x 10(-2) m) and calcium chloride (CaCl2; 10(-4)-10(-2)) were isometrically recorded at 37 and 41 degrees C (warming). During warming the sensitivity, but not the maximal response, of carbachol 5-HT, KCl, and CaCl2 was significantly higher than at 37 degrees C. Warming to 41 degrees C after treatment with NG-nitro-L arginine methyl esther (10(-5) m) did not modify the effect of warming. These results suggest that nitric oxide seems to have no role in the warming-induced responses in calf cardiac vein. PMID- 12588634 TI - Sympatho-inhibitory actions of irbesartan in pithed spontaneously hypertensive and Wistar-Kyoto rats. AB - Angiotensin II (Ang II) can enhance sympathetic neurotransmission by acting on (AT1) receptors that are located on sympathetic nerve terminals. We investigated presynaptic blockade by the selective AT1-receptor antagonist irbesartan in pithed spontaneously hypertensive rats and normotensive Wistar-Kyoto rats (WKY). We compared the presynaptic inhibitory dose with that required for the blockade of AT1-receptors on vascular smooth muscle in both strains. To investigate blockade of presynaptic AT1-receptors, we studied the effect of irbesartan on the sequelae of electric stimulation of the thoraco-lumbar sympathetic outflow (0.25 8 Hz). To study the interaction between postsynaptic AT1-blockers and alpha adrenoceptors, the effects of irbesartan on pressor responses to exogenous noradrenaline (NA) were established. Additionally, we studied the effect of irbesartan on dose-response curves for the vasoconstriction induced by exogenous Ang II. Pressor responses to electrical stimulation of thoracolumbar sympathetic neurones, to exogenous Ang II, as well as to (NA) were enhanced in spontaneously hypertensive rats (SHR) compared with WKY. The stimulation-induced rise in DBP could be dose-dependently reduced by irbesartan (0.3-10 mg/kg) in both SHR and WKY. The pIC50 values (doses which suppress the rise in DBP by 50% compared with control) were 5.60 +/- 0.09 and 5.72 +/- 0.08 for SHR and WKY, respectively (P > 0.05). In SHR, no effect of irbesartan (3 mg/kg) on pressor responses to exogenous NA was observed. In contrast, in WKY, irbesartan (3 mg/kg) caused a rightward shift of the dose-response curve to exogenous NA. Irbesartan (0.3-3 mg/kg) caused a depression of E(max) values and a rightward shift of the dose response curves to exogenous Ang II in a similar fashion in both SHR and WKY. From these results we conclude that both in SHR and in WKY, Ang II exerts a facilitatory effect on sympathetic neurotransmission, which is mediated by prejunctional AT1-receptors in both strains. Irbesartan displays comparable sympatho-inhibitory potency in the normotensive and hypertensive pithed rat preparations. A facilitatory effect via postsynaptically located AT1-receptors on alpha-adrenoceptor-mediated responses exists in WKY, but not in SHR. In both strains the required dose to inhibit presynaptic effects is somewhat higher than the dose required to inhibit postsynaptic effects. No differences, therefore, seem to exist between the two strains regarding the affinity of irbesartan for pre- and postjunctional AT1-receptors, respectively. PMID- 12588635 TI - A clinical pharmacological study of the potential beneficial effects of a propolis food product as an adjuvant in asthmatic patients. AB - The aqueous extract of propolis has been formulated as a nutritional food product and administered, as an adjuvant to therapy, to patients with mild to moderate asthma daily for 2 months in the framework of a comparative clinical study in parallel with a placebo preparation. The diagnosis of asthma was made according to the criteria of patient classification of the National Institutes of Health and Global Initiative for Asthma Management. At inclusion, the pulmonary forced expiratory volume in the first second (FEV1) as a percentage of the forced vital capacity (FVC) was more than 80% in mild persistent cases, and between 60 and 80% in moderate persistent cases, showing an increase in the degree of reversibility of > 15% in FEV1. All patients were on oral theophylline as controller therapy, none was receiving oral or inhaled corticosteroids, none had other comorbidities necessitating medical treatment, and all were from a middle-class community and had suffered from asthma for the last 2-5 years. Twenty-four patients received the placebo, with one drop-out during the study, while 22 received the propolis extract, with no drop-outs. The age range of the patients was 19-52 years; 36 were male and 10 female. The number of nocturnal attacks was recorded on a weekly basis, while pulmonary function tests were performed on all patients at the beginning of the trial, 1 month later and at the termination of the trial. Immunological parameters, including various cytokines and eicosanoids known to play a role in asthma, were measured in all patients at the beginning of the trial and 2 months later. Analysis of the results at the end of the clinical study revealed that patients receiving propolis showed a marked reduction in the incidence and severity of nocturnal attacks and improvement of ventilatory functions. The number of nocturnal attacks dropped from an average of 2.5 attacks per week to only 1. The improvement in pulmonary functions was manifested as a nearly 19% increase in FVC, a 29.5% increase in FEV1, a 30% increase in peak expiratory flow rate (PEFR), and a 41% increase in the forced expiratory flow rate between 25 and 75% of the vital capacity (FEF25-75). The clinical improvement was associated with decreases by 52, 65, 44 and 30%, respectively, of initial values for the pro-inflammatory cytokines tumor necrosis factor (TNF) alpha, ICAM-1, interleukin (IL)-6 and IL-8, and a 3-fold increase in the 'protective' cytokine IL-10. The levels of prostaglandins E2 and F2alpha and leukotriene D4 were decreased significantly to 36, 39, and 28%, respectively, of initial values. Patients on the placebo preparation showed no significant improvement in ventilatory functions or in the levels of mediators. The findings suggest that the aqueous propolis extract tested is potentially effective as an adjuvant to therapy in asthmatic patients. The benefits may be related to the presence in the extract of caffeic acid derivatives and other active constituents. PMID- 12588636 TI - Instant centre frequency at anaesthetic induction--a new way to analyse sympathovagal balance. AB - The instant centre frequency (ICF) of RR interval has been proposed as a global index to analyse the sympathovagal interaction in the heart. The aim of this study was to assess the ICF during anaesthesia to test if it can reliably capture the neural control of the cardiovascular system. Twenty-four ASA II or III patients scheduled for cardiac surgery were included in the study. They were allocated in two groups: control, no treatment (group 1, n = 12), and beta adrenergic blockade by atenolol (group 2, n = 12). Spectra of pulse interval series were computed with a time-frequency method and they were divided into: very low frequency (VLF, 0.000-0.040 Hz), low frequency (LF, 0.050-0.150 Hz) and high frequency (HF, 0.160-0.500 Hz). Normalized power was obtained by dividing the cumulative power within each frequency band (LF or HF) by the sum of LF and HF; the ratio of LF/HF was also calculated. Instant centre frequency is a time varying parameter that the evolution along time of the gravity centrum of a local spectrum. All spectral indexes were recorded at the following time points: before induction, after induction and before intubation, during intubation, and after intubation. The atenolol group had lower normalized LF and the LF/HF ratio (P < 0.05) higher HF before induction; and lower LF/HF ratio after induction and before intubation (P < 0.05). The ICF was higher in atenolol group at all times. The ICF shifted towards HF frequency after induction and before intubation and shifted towards LF during intubation in both groups. The autonomic nervous system control on the heart through the interaction of sympathetic and parasympathetic reflex mechanisms could be studied by the ICF. The ICF may assess the autonomic cardiac modulation and may provide useful information for anaesthetic management. PMID- 12588637 TI - Excessive daytime sleepiness and 'sleep attacks' induced by entacapone. AB - Three patients with advanced Parkinson's disease, all of whom developed excessive daytime sleepiness and 'sleep attacks' after the administration of entacapone, are described. This is another demonstration that inappropriate daytime sleep episodes are not exclusive to dopamine agonists. PMID- 12588638 TI - Piribedil-induced sleep attacks in Parkinson's disease. AB - 'Sleep attacks', episodes of sudden onset of sleep without any prodromal symptoms, were initially described in patients with Parkinson's disease (PD) taking the newer dopamine agonists pramipexole and ropinirole. Piribedil, a nonergot agonist with both D2 and D3 agonist action, is an effective antiparkinsonian medication. However, there are very few reports of Piribedil induced sleep attacks in PD. Among 50 PD patients seen at our Movement Disorder Clinic who had recently taken Piribedil, we identified three (6%) who satisfied the clinical description of sleep attacks. Here we provide details of the clinical characteristics of Piribedil-induced sleep attacks in these PD patients. PMID- 12588639 TI - Sleep attacks in Parkinson's disease induced by Entacapone, a COMT-inhibitor. AB - We present a patient who suffered from sleep attacks after starting entacapone in addition to levodopa. Entacapone, a catechol-O-methyl transferase inhibitor, alters the pharmacokinetics of levodopa, leading to increase of levodopa concentration in plasma and brain. This mechanism is suspected to be involved in the pathophysiology of sleep attacks in this case. PMID- 12588640 TI - Unlicensed and off-label uses of drugs in paediatrics: a review of the literature. AB - The off-label and unlicensed use of drugs to treat children is a common practice that occurs either in hospital or in the community. This derives from the fact that research for establishing drug efficacy and safety in children has not been carried out due to ethical problems, logistical difficulties, financial and legal concerns. In this work we report the studies available in literature documenting the extent of drug use in the paediatric field outside the recommendations of the license. From our analysis, a widespread attitude to prescribe medicines to children outside their product license either in the hospitals or in the community is confirmed. This suggests an immediate action for a more rationale use of drugs in paediatrics, to avoid exposing children and infants to unnecessary risks, but also to avoid depriving them of potentially effective and sometimes life-saving therapies. PMID- 12588642 TI - Cellular implants: pioneers in xenotransplantation? PMID- 12588643 TI - B cell tolerance to xenoantigens. AB - Xenotransplantation of pig organs to humans is a possible solution to the shortage of donor organs for transplantation. Multiple immunologic barriers need to be overcome if pig-to-primate transplantation is to be successful. The presence, in humans, of natural antibodies (Abs) directed against Galalpha1 3Galbeta1-4GlcNAc epitopes on pig vascular endothelium provides the major barrier, as antibody-antigen binding initiates the process of hyperacute rejection. Even if hyperacute rejection is prevented, acute vascular rejection develops. Acute vascular rejection is also mediated, in part, by xenoreactive Abs and may be complement-independent. Efforts being made to overcome antibody mediated rejection include depletion of antibody by extracorporeal immunoadsorption, prevention of an induced Ab response by pharmacologic reagents, B-cell and/or plasma cell depletion, depletion or inhibition of complement, and the use of organs from pigs transgenic for human complement regulatory proteins. The ultimate solution would be the induction of B-cell tolerance to xenogeneic antigens, which is being explored by attempting to induce xenogeneic hematopoietic chimerism. Here, we review the properties of the B cell types responding to xenoantigens and the strategies for tolerizing those B cells. PMID- 12588644 TI - Analysis of human CD4 T lymphocyte proliferation induced by porcine lymphoblastoid B cell lines. AB - BACKGROUND: This study was undertaken to characterize the two porcine lymphoblastoid cell lines L23 and L35, derived from a pig inoculated by the retrovirus Tsukuba-1, and to determine how they induce a strong human lymphocyte proliferation. METHODS: Phenotypic characterization was performed by flow cytometry and reverse transcriptase-polymerase chain reaction analyses. Xenogeneic mixed lymphocyte reactions (XMLR) were performed using unfractionated human peripheral blood mononuclear cells (huPBMC) and purified CD4+ T lymphocytes as responding cells, in the presence of blocking antibodies and fusion proteins. RESULTS: The immunoglobulin genes were demonstrated to be rearranged in L23 and L35 cell lines, in agreement with the expression of a B cell phenotype. Both induced a similar proliferation of huPBMCs and purified human CD4+ lymphocytes from adult or cord blood (naive cells). Proliferation of CD4+ T lymphocytes was completely blocked by anti-SLA-DR plus anti-SLA-DQ mAbs, excluding human lymphocyte transformation by porcine viruses. The frequency of proliferative precursors was inconsistent with that induced by a retroviral superantigen but similar to classical direct xenoantigen presentation as observed with other porcine antigen-presenting cells. Extensive analysis of costimulatory signals led to the identification of the CD28 pathway, in agreement with membrane expression of B7 molecules on L23 and L35 cells, and of the CD2 pathway in L35 cells. CONCLUSION: These two porcine lymphoblastoid cell lines have been further characterized and clearly identified as belonging to the B cell lineage. By expressing major histocompatibility complex class II antigens and costimulatory molecules, they induce a vigorous proliferative response of human CD4+ lymphocytes through a direct presentation pathway. PMID- 12588645 TI - Hyperacute lung rejection in the pig-to-human model. 2. Synergy between soluble and membrane complement inhibition. AB - BACKGROUND: The role of complement in hyperacute lung xenograft rejection has not been elucidated. The present study evaluates the effect of complement (C) C3/C5 convertase inhibition on hyperacute rejection of pig lung by human blood. METHODS: In an established ex-vivo model, lungs from pigs heterozygous for human decay accelerating factor (hDAF), non-transgenic littermate control pigs, or farm bred pigs were perfused with fresh human blood that was either unmodified or treated with soluble complement receptor type 1 (sCR1: TP10, 100 microg/ml). RESULTS: Non-transgenic lungs from littermate controls had a median survival time of 35 min (range 5 to 210; P = 0.25 vs. farm-bred piglets: median 5 min, range 5 to 10). Lungs expressing hDAF survived for a median of 90 min (range 10 to 161; P = 0.5 and 0.01 vs. littermate and farm-bred controls, respectively), with sCR1, whereas hDAF (-) lungs failed by 35 min (range 6 to 307), hDAF (+) lungs survived for 330 min (range 39 to 577) [P = 0.002 vs. farm-bred; P = 0.08 vs. hDAF (-); P = 0.17 vs. sCR1/hDAF (-)]. The rise in pulmonary vascular resistance (PVR) at 5 min was blunted only by hDAF (+) with sCR1 (0.26 +/- 0.2 vs. 0.5 to 0.7 mmHg/ml/min for other groups). Plasma C3a and sC5b-9 and tissue deposition of C5b 9 were dramatically diminished using sCR1, and further decreased in association with hDAF. Histamine and thromboxane were produced rapidly in all groups. CONCLUSION: Complement plays an important role in lung HAR. However, even potent inhibition of C3/C5 convertase, both membrane bound in lung and by a soluble phase inhibitor in the blood, does not prevent activation of inflammatory responses known to be particularly injurious to the lung. Our findings implicate a role for innate immune pathways resistant to efficient complement regulation. The role of anti-species antibody, coagulation pathway dysregulation, and additional environmental or genetic influences remain to be defined. PMID- 12588646 TI - Effect of various forms of the C1 esterase inhibitor (C1-INH) and DAF on complement mediated xenogeneic cell lysis. AB - The purpose of the present study was to assess the effect of various forms of the surface-bound form of the C1 esterase inhibitor (C1-INH-PI) and decay accelerating factor (DAF) on xenogenic cells. cDNAs of various deletion mutants of the C1-INH-PI, such as delta-1-99 amino acid (AA), delta-108-183AA loop, delta whole loop, delta-exon5, delta-exon6 + 7, and delta-exon5 + 6 + 7, and that of DAF, the delta-short consensus repeat (SCR) 1-DAF were established. While all deletion mutants of C1-INH-PI except the delta-1-99AA were expressed in the cytoplasm but not on the cell surface, the delta-1-99AA was clearly expressed on the xenogeneic cell surface. Amelioration of complement-mediated xenogeneic cell lysis by delta-1-99AA was next tested, and compared with delta-SCR1 DAF. Both molecules blocked human complement-mediated cell lysis by approximately 57 to 90 and 93 to 98%, respectively, in Chinese hamster ovarian tumor (CHO) cells and pig endothelial cells (PECs). The CHO cell transfectants were incubated with 20% normal human serum, and the amounts of C4 and C3 deposition on the cell surface were analysed by flow cytometry. The DAF transfectant showed a large amount of C4 deposition and much less C3-deposition than the controls (approximately 85% suppression), whereas the delta-1-99AA showed approximately a 40% suppression in both C4- and C3-deposition. Consequently, both the delta-1-99AA C1-INH-PI and delta-SCR1 DAF molecules are quite effective in down-regulating the xenogeneic cell lysis, but accomplished this in different manners. PMID- 12588647 TI - Porcine cytomegalovirus in pigs being bred for xenograft organs: progress towards control. AB - In human medicine, human cytomegalovirus (HCMV) is readily transmitted by organ transplant causing end-organ disease and triggering graft rejection in recipients. Because of a chronic shortage of human organs, pigs transgenic for human complement control proteins are being considered as potential donors. Such xenotransplantation raises concerns about the potential zoonotic transmission of viruses including porcine cytomegalovirus (PCMV), an endemic infection of pigs. Similar to HCMV and PCMV transmission is thought to occur in utero and perinatally. We used quantitative polymerase chain reaction to examine the prevalence, organ distribution and viral load of PCMV in human decay accelerating factor (CD55) transgenic pigs. In animals reared under conventional farm conditions, virus was identified in a wide range of organs including potential xenografts (liver, kidney and heart). The spleen was PCMV DNA positive in all infected pigs. Examination of foetal spleens failed to identify evidence of transplacental infection and prospective monitoring of two litters showed that infection occurred in the postnatal period. This transmission was prevented by hysterotomy derivation and barrier rearing. Our findings demonstrate that PCMV could be eradicated from pig herds being bred for xenotransplantation and argue that the spleen from donor animals should be examined as part of quality control procedures if clinical trials proceed. PMID- 12588648 TI - Multivalent Galalpha1,3Gal-substitution makes recombinant mucin-immunoglobulins efficient absorbers of anti-pig antibodies. AB - Hyperacute organ xenograft rejection can be prevented by removing anti-pig antibodies by extracorporeal absorption prior to transplantation. A novel recombinant absorber of anti-pig antibodies was developed by fusing the cDNA encoding the extracellular part of a mucin-type protein, P-selectin glycoprotein ligand-1, with an antibody Fc fragment cDNA, which upon coexpression with the porcine alpha1,3 galactosyltransferase carried the xenogeneic epitope, Galalpha1,3Gal (Liu J., Qian Y., Holgersson J., Transplantation 1997, 63, 1673 1682). The biochemical characterization of the mucin/Ig and its absorption efficacy compared with that of porcine thyroglobulin and Galalpha1,3Gal conjugated beads are reported. The carbohydrate portion of the mucin/Ig constituted 43% of its molecular weight and the majority of the Galalpha1,3Gal epitopes were O-linked as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting following N-glycosidase F digestion. Gas chromatography-mass spectrometry of reduced and acetylated saccharides released by alpha-galactosidase treatment revealed that the fusion protein carried approximately 140 mol of terminal, alpha-linked galactose per mole protein. Based on the reduction in pig aortic endothelial cell cytotoxicity, Galalpha1,3Gal substituted mucin/Igs on agarose beads were, on a carbohydrate molar basis, shown to be approximately 20 times more efficient than agarose-conjugated pig thyroglobulin, and approximately 5000 and 30,000 times more efficient than Galalpha1,3Gal-substituted agarose and macroporous glass beads, respectively. Structural features of the mucin backbone and its carbohydrate core saccharide chains determine the structural context, spatial orientation and spacing of Galalpha1,3Gal epitopes and are likely to explain the superior absorption efficacy of the recombinant mucin-type chimera. PMID- 12588649 TI - The role of chemokines and their receptors in the rejection of pig islet tissue xenografts. AB - The mechanism by which inflammatory cells are recruited to pig islet tissue (proislet) xenografts was investigated by examining the intragraft mRNA expression of murine alpha- and beta-chemokines in CBA/H mice from days 3 to 10 post-transplant. Xenograft rejection was associated with early intragraft transcript expression for monocyte chemotactic protein-1 (MCP-1) (3 to 5 days), IP-10 (3 to 4 days) and macrophage inflammatory protein-1alpha (MIP-1alpha) (3 to 5 days) and subsequent expression of eotaxin (days 4 to 10), MIP-1beta (days 4 and 5) and regulated on activation, normal T cell expressed and secreted (RANTES) (days 4 to 6) mRNA. This pattern was consistent with the early recruitment of macrophages (MCP-1, MIP-1alpha), the influx of CD4 T cells (MCP-1, MIP-1alpha, MIP-1beta, IP-10 and RANTES) and the characteristic infiltrate of eosinophils (eotaxin and RANTES) associated with islet xenograft rejection. Inhibition of beta-chemokine signaling in CCR2-/- mice (which lack the major co-receptor for MCP-1) resulted in retarded macrophage and CD4 T cell recruitment, enhanced eosinophil influx and a minor delay in rejection, compared with wildtype mice; there was little effect on leukocyte infiltration in xenografts harvested from CCR5-/- mice (lacking the co-receptor for MIP-1alpha, MIP-1beta and RANTES). The impeded migration of leukocytes into xenografts in CCR2-/- hosts was associated with delayed intragraft expression of MCP-1 and RANTES mRNA; absence of MCP 1/CCR2-mediated signaling led to enhanced intragraft expression of MCP-1, MIP 1alpha and MIP-1beta mRNA. These findings suggest that MCP-1 plays an important role in regulating macrophage and CD4 T cell infiltration to xenograft sites via the CCR2 signaling pathway. Additional treatment of xenografted CCR2-/- transplant recipients with anti-interleukin-(IL)-4 and anti-IL-5 mAbs further delayed xenograft rejection demonstrating the potential for combined antirejection strategies in facilitating pig islet xenotransplantation. PMID- 12588650 TI - Pig islet xenografts are resistant to autoimmune destruction by non-obese diabetic recipients after anti-CD4 treatment. AB - In addition to providing a large source of donor tissue, xenogeneic islet transplantation might avoid recurrent autoimmunity in patients with type 1 diabetes. To examine this possibility further, xenogeneic pig islets were transplanted into recipient mice in the presence or absence of autoimmunity. Spontaneously, non-obese diabetic (NOD) recipients rejected isografts rapidly whether or not the recipients were depleted of CD4+ T-cells. Young NOD mice made diabetic with streptozotocin accepted islet isografts without immunosuppression, indicating that destructive autoimmunity did not develop in these recipients. Pig xenografts were rejected equally quickly in the two types of NOD recipients in the absence of immunosuppression and survived for up to 9 weeks in both types of NOD recipients after CD4 depletion. BALB/c mice often accepted pig xenografts indefinitely after anti-CD4 antibody treatment. These results suggest that pig islets are resistant to recurrent autoimmunity when CD4+ T-cells are depleted. The difficulty in obtaining indefinite islet xenograft survival in NOD recipients occurs independently from the development of destructive autoimmunity. PMID- 12588651 TI - Graft thrombosis in hDA-transgenic pig hearts transplanted into rhesus monkeys. PMID- 12588652 TI - Neurotoxicity of leflunomide in baboons. PMID- 12588654 TI - Calcineurin phosphatase activity and immunosuppression. A review on the role of calcineurin phosphatase activity and the immunosuppressive effect of cyclosporin A and tacrolimus. AB - The mode of immunosuppressive action of tacrolimus (FK506) and cyclosporin A has been elucidated. Both drugs bind to proteins in the cytoplasm to form complexes, which in turn inhibit the phosphatase activity of calcineurin, an important limiting step in the activation of T cells. The association between drug uptake (pharmacokinetics) and enzyme inhibition (pharmacodynamics) is under current investigation. Great variations in the correlation between blood drug levels and enzyme inhibition could indicate that monitoring calcineurin phosphatase activity for treatment might be superior to monitoring blood drug levels. PMID- 12588655 TI - Pharmacological disintegration of lipid rafts decreases specific tetramer binding and disrupts the CD3 complex and CD8 heterodimer in human cytotoxic T lymphocytes. AB - Accumulating evidence strongly supports the role of lipid rafts in the regulation of T-lymphocyte activation, but the organization and molecular composition of these cholesterol- and sphingolipid-rich membrane microdomains in different subsets of T cells remain poorly investigated. Here, we show that pharmacological disruption of lipid rafts in human CD8+ cytotoxic T-lymphocyte (CTL) clones disturbs the integrity of CD3 complex and CD8 heterodimer, without affecting the reactivity with T-cell receptor (TCR)-specific antibodies. This demonstrates that interaction with completely assembled CD3 complex is not required for the stable expression of TCR at the cell surface. The effect of raft disruption on CD3 and CD8 expression correlates with failure to bind specific tetrameric complexes by a proportion of surface TCR molecules. However, the interaction of specific tetramer with the rest of surface TCR pools appears to be unaffected, demonstrating that TCR-signalling complexes may differ in their requirement for cholesterol to stably maintain their composition and to rearrange for efficient tetramer binding. Together with previously published data, our results support the existence of molecular and/or structural heterogeneity of lipid rafts that may play an important role in controlling distinct functional properties of T cell subsets. PMID- 12588656 TI - Transfer of macrophage-derived mycobacterial antigens to dendritic cells can induce naive T-cell activation. AB - Mycobacteria are capable of surviving and replicating in host macrophages, where they can release antigenic material into the environment. However, unlike dendritic cells (DCs), macrophages do not appear to be capable of activating naive T cells. Therefore, this work investigated antigen transfer between macrophages and DCs. We generated culture supernatants from bacille Calmette Guerin (BCG)-infected and uninfected macrophages and then determined whether DCs could present these extracellular mycobacterial antigens to T cells. Here, we show that DCs pulsed with antigens released from BCG-infected macrophages can stimulate primed T cells in vitro and initiate naive T-cell responses in vivo. These results suggest that antigen transfer can occur between macrophages and DCs. PMID- 12588657 TI - Cytokine production and expression of leucocyte-differentiation antigens by human mononuclear cells in response to mycobacterium tuberculosis antigens. AB - The aim of this work was to characterize a leucocyte-differentiation antigen or chemokine receptor that allows the identification of type 1 (T helper 1 (Th1), Tc1) and type 2 (Th2, Tc2) lymphocytes in short-term-cultured human peripheral blood mononuclear cells. In addition, we assessed the type of response induced by mycobacterial antigens in tuberculosis patients and healthy contacts. Cells were stimulated with an unfractionated culture filtrate or 30 kDa antigen from Mycobacterium tuberculosis. Then, CD4 and CD8 cell labelling was combined with CD30, CD27, CD28, CD45RA or CD45R0 staining, detection of intracellular interferon-gamma (IFN-gamma) or interleukin-4 (IL-4) and analysis by three-colour flow cytometry. In separate experiments, the expression of different chemokine receptors (CCR1, CCR3, CCR5, CXCR3 and CXCR4) was also studied. We found that none of the cell-surface molecules studied was preferentially expressed by Th1 or Th2 cells. Thus, our results indicate that these lymphocyte subsets cannot be identified in short-term-cultured mononuclear cells on the basis of preferential expression of the cell markers studied, and that it is necessary to look for additional molecules that allow the discrimination of Th1 and Th2 cells. PMID- 12588658 TI - Human Th1 cell lines recognize the Mycobacterium tuberculosis ESAT-6 antigen and its peptides in association with frequently expressed HLA class II molecules. AB - We have used a synthetic-peptide approach to map epitope regions of the Mycobacterium tuberculosis ESAT-6 antigen recognized by human T cells in relation to major histocompatibility complex (MHC) restriction. ESAT-6-specific CD4+ T cell lines were established by stimulating peripheral blood mononuclear cells from 25 HLA-DR-typed tuberculosis patients with complete antigen in vitro. The established T-cell lines were then screened for proliferation and interferon gamma (IFN-gamma) secretion in response to eight overlapping 20-mer peptides covering the ESAT-6 sequence. The response of the T-cell lines to ESAT-6 and peptides from a human leucocyte antigen (HLA)-heterogeneous group of donors suggested the presence of multiple epitopes and promiscuous recognition of the antigen. Analysis of antigen and peptide recognition in the presence of anti-HLA class I and class II antibodies suggested that the T-cell lines recognized ESAT-6 in association with HLA-DR and -DQ molecules. Furthermore, testing of selected T cell lines with ESAT-6 and the peptides in the presence of autologous and allogeneic HLA-DR- and -DQ-typed antigen-presenting cells identified HLA-DR2, DR52 and -DQ2 amongst the HLA molecules involved in the presentation of ESAT-6 and its peptides to human Th1 cells. In addition, the T-cell lines were cytotoxic for monocytes and macrophages pulsed with ESAT-6 and peptides. In conclusion, the recognition of ESAT-6 by IFN-gamma-secreting and cytotoxic CD4+ T cells in association with frequently expressed HLA class II molecules supports the application of this antigen to either specific diagnosis or subunit vaccine design. PMID- 12588659 TI - Differences in chicken major histocompatibility complex (MHC) class Ialpha gene expression between Marek's disease-resistant and -susceptible MHC haplotypes. AB - The expression of chicken major histocompatibility complex (MHC) class Ialpha genes was investigated in spleen cells from a panel of chickens with well-defined MHC haplotypes, and two class Ialpha transcripts of 1.9 and 1.5 kb were detected in various amounts. In BW1, B130 and B21, the two transcripts were almost equally expressed. In B2, B6, B12 and B19, the ratio between the two transcripts was 4 : 1, with the 1.9 kb transcript having the strongest expression. In B14 and B15, the 1.5 kb transcript was undetectable and the 1.9 kb transcript appeared to be exclusively expressed. Thus, haplotypes considered to have an MHC-determined resistance to Marek's disease (MD) had the highest relative amount of the 1.5 kb transcript, whereas haplotypes considered to be MD-susceptible had the lowest. In order to address a possible correlation between MHC-Ialpha transcriptional patterns and MD resistance, a larger animal material experimentally infected with MD virus (MDV) was examined. The expression of MHC class Ialpha genes was investigated in spleens as well as in other organs, 9 weeks post-infection (p.i.), from animals of the two MD-resistant haplotypes B21 and BW1 as well as from the MD-susceptible haplotype B19. In the spleen cells of infected animals, the relative amount of the 1.5 kb transcript in the haplotypes BW1 and B21 was shown to be significantly higher than that in B19. Interestingly, in infected BW1 and B21 animals, the relative amount of the 1.5 kb transcript was also significantly higher than that in healthy MHC-matched controls. In B19, no differences were detected between uninfected and infected animals. Furthermore, it was shown in BW1 and B21 that the two classical MHC-Ialpha genes located in the MHC region were both able to produce both mRNA transcripts. Hybridization experiments, using specific probes upstream and downstream of the polyadenylation signals in the 3' end of the MHC-Ialpha genes, demonstrated that alternate use of these signals is probably involved in the production of the two mRNA transcripts. PMID- 12588660 TI - Effects of various adjuvants and a viral infection on the antibody specificity toward native or cryptic epitopes of a protein antigen. AB - An immunization protocol that induces antibodies (Abs) directed to cryptic epitopes of a protein antigen (Ag) reduces the efficacy of vaccines that ideally should induce Abs against native epitopes. We have shown earlier that viral infections concomitant with immunization against a protein tend to shift the Ab specificity toward cryptic epitopes and tend to induce the production of autoantibodies (autoAbs). Here, we show the effects of three adjuvants on the Ab specificity in the absence or presence of a viral infection (lactate dehydrogenase-elevating virus or LDV), with human growth hormone (hGH) being, as before, the protein Ag. Pathogen-free CBA/Ht and BALB/c mice were immunized with hGH in the presence of complete Freund's adjuvant (CFA), monophosphoryl lipid A (MPL) or alum, with the animals being either infected with LDV or not infected with LDV. Conventional and competition enzyme-linked immunosorbent assays (ELISAs) indicated that in noninfected mice, CFA induced higher titres of anti hGH Ab than did MPL or alum, with the Ab being almost totally directed to cryptic hGH epitopes. Strikingly, CFA plus LDV infection in CBA/Ht mice shifted the specificity of the anti-hGH Ab toward native epitopes, whereas the virus decreased the Ab titre when MPL or alum was used. Our Western blot results showed that 70% of mice immunized with hGH in the presence of any adjuvant produced autoAbs against a variety of tissue Ags. The amount of autoAb and the concentration of Ab to hGH cryptic epitopes did correlate, suggesting a relationship between both kinds of Ab. Significant differences were observed in the various effects of adjuvants and the viral infection between the two mouse strains used in this work. PMID- 12588661 TI - T- and B-cell epitopes in the secreted Mycobacterium bovis antigen MPB70 in mice. AB - MPB70 is a soluble secreted protein highly expressed in Mycobacterium bovis and strains of bacille Calmette-Guerin (BCG); as such, it is a candidate for subunit and DNA vaccines against tuberculosis. MPB70 was screened for T-cell epitopes in four different inbred mouse strains. Major histocompatibility complex (MHC) H-2b expressing mice (C57BL/6) secreted interferon-gamma (IFN-gamma) after stimulation with peptides from the regions 1-20, 41-50, 81-110, 121-150 and 161-193 of the MPB70 sequence. H-2db mouse (B6D2) splenocytes secreted IFN-gamma after stimulation with some of the same peptides, whereas H-2d mice (BALB/c and DBA/2) did not secrete IFN-gamma upon stimulation with the peptides. Sera from H-2db mice immunized with native MPB70 in incomplete Freund's adjuvant (IFA), mpb70 DNA or live BCG Moreau were found to contain antibodies against the native MPB70 antigen. H-2db mice immunized with native MPB70 in IFA exhibited high titres of peptide-reactive immunoglobulin G1 (IgG1) antibodies, whereas DNA-immunized mice reacted with IgG2a antibodies against some of the same peptides. As some of the epitopes recognized by mouse T and B cells have previously been found to stimulate immune responses in humans, cattle and rabbits, we conclude that these epitopes may be good general epitopes for the stimulation of T- and B-cell responses and candidates for a DNA vaccine with a broad applicability. PMID- 12588662 TI - Effect of flavone derivatives on interleukin-1beta (IL-1beta) mRNA expression and IL-1beta protein synthesis in stimulated RAW 264.7 macrophages. AB - It is known that the redox status of cells affects gene expression. Flavones, as natural antioxidants, efficiently modulate this status and may play a role in the regulation of inducible gene expression of inflammatory mediators. This study was designed to investigate the effect of five flavone derivatives variously substituted with hydroxyl groups (chrysin, galangin, kaempferol, quercetin and myricetin) on interleukin-1beta (IL-1beta) gene expression in stimulated RAW 264.7 macrophages. The cells were incubated with tested hydroxyflavones and stimulated with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). Then, the following were estimated: the level of IL-1beta mRNA in these cells and the concentration of IL-1beta protein in cell-culture supernatants and cell lysates. Each of the tested compounds significantly decreased IL-1beta mRNA expression. The most potent inhibitor was chrysin (hydroxyflavone with two hydroxyl groups and a weak antioxidant activity). The effects of galangin and kaempferol were similar. Myricetin (hydroxyflavone with a strong antioxidant activity) significantly decreased the level of IL-1beta mRNA, but it had no effect on the IL-1beta protein synthesis. The results indicated that hydroxyflavones could modulate the IL-1beta gene expression in activated RAW 264.7 macrophages via inhibiting gene transcription. This action seems unlikely to be the result of antioxidant properties of tested compounds. PMID- 12588663 TI - Umbilical cord mannan-binding lectin and infections in early childhood. AB - To investigate whether mannan-binding lectin (MBL) in umbilical cord blood (UCB) is associated with the incidence rate of hospitalizations for infections during early childhood. A follow-up study from birth to 31 months of age, with endpoint data from the Danish National Hospital Discharge Registry. The concentration of MBL and immunoglobulin A (IgA) in UCB was measured in a time-resolved immunofluorometric assay. Information on possible confounding factors was obtained from questionnaires completed by mothers during their pregnancy, and characteristics of the child at birth were registered by a midwife on a structured coding sheet. A total of 2104 infants born in the period between 1 February 1990 and 25 May 1991 to mothers living in the municipality of Aarhus. Concentration of MBL and IgA in UCB, gestational age, birth weight, number of siblings less than 3 years old, mother's smoking habits, alcohol and coffee consumption, educational and marital status and previous spontaneous abortions and stillbirths. Of 2104 children, 626 were hospitalized at least once, and 346 of these were hospitalized with infection. The hazard ratio (HR) of hospitalization with infections in children with low levels of MBL (<120 ng/ml) was 1.4 (95% confidence interval (CI), 1.0-1.8), which was similar to our findings on hospitalization because of other diseases. The HR of hospitalization for viral infections was 2.8 (CI, 1.3-5.9). Low levels of MBL in UCB were associated with hospitalization in general and particularly with hospitalization for viral infections. PMID- 12588664 TI - Cytokines in patients with lung cancer. AB - Lung cancer is one of the most common malignant diseases and is amongst the leading causes of death. Cell-mediated immune response and cytokines could play an important role in antitumour immunity. The aim of the study was to evaluate the cytokines', tumour necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL 1beta) and IL-6, releasing capacity in patients with lung carcinoma and benign lung disease. A group of 41 patients were tested for the production of TNF-alpha, IL-1beta and IL-6 in bronchoalveolar lavage (BAL) and blood. The levels of cytokines in the lung cancer patients were: (1) in BAL - IL-6, 173 +/- 85 pg/ml; TNF-alpha, 170 +/- 116 pg/ml; and IL-1beta, 473 +/- 440 pg/ml; (2) in the blood - IL-6, 197 +/- 53 pg/ml; TNF-alpha, 311 +/- 202 pg/ml; and IL-1beta, 915 +/- 239 pg/ml. Alveolar macrophages of the patients with a lung cancer secreted significantly more cytokines, IL-6 (P = 0.0004) and IL-1beta (P = 0.0047), than alveolar macrophages of the patients with a nonmalignant lung cancer. However, significantly lower levels of cytokine production by the BAL cells were found in patients with small cell lung cancer. This production decreased further in phase IV of nonsmall cell lung cancer. PMID- 12588665 TI - Antineutrophil cytoplasmic antibodies induce decreased CD62L expression and enhanced metabolic activity in monocytes. AB - Monocyte in vitro activation by antimyeloperoxidase (anti-MPO)- and antiproteinase-3 (anti-PR3)-positive sera, corresponding immunoglobulin G (IgG) fractions and monoclonal antibodies against MPO and PR3 was evaluated. The expression of adhesion molecules, l-selectin (CD62L) and CR3 (CD11b), involved in leucocyte endothelial adhesion, and metabolic activity, measured as the production of hydrogen peroxide, were analysed. Decreased expression of CD62L was demonstrated in monocytes after incubation with antineutrophil cytoplasmic antibody (ANCA)-positive sera. This finding was not accompanied by changes in CD11b expression. Metabolic activity was increased in monocytes after incubation with ANCA-positive IgG fractions as well as after incubation with monoclonal anti MPO and anti-PR3. These findings support the concept that the pathophysiological effect of ANCA is partly mediated through the action on crucial events in monocyte activation, such as CD62L downregulation and oxygen radical production. PMID- 12588666 TI - Indoleamine 2,3-dioxygenase expression in patients with acute graft-versus-host disease after allogeneic stem cell transplantation and in pregnant women: association with the induction of allogeneic immune tolerance? AB - Indoleamine 2,3-dioxygenase (IDO) is an interferon-gamma (IFN-gamma)-induced enzyme, which is suggested to play an important role in the prevention of allogeneic fetal rejection. IDO effects the suppression of T-cell activity by catabolizing the essential amino acid l-tryptophan. We studied IDO expression by reverse transcription polymerase chain reaction (RT-PCR) in dendritic cells and by real-time RT-PCR in monocytes of patients undergoing allogeneic transplantation for leukaemia, who developed acute graft-versus-host disease (aGvHD), and compared the IDO expression with that of pregnant women and healthy volunteers. A spontaneous IDO expression was detected in the monocytes of 20 pregnant women with an IDO/glyceraldehyde-3-phosphate dehydrogenase (GAPDH) ratio at a median of 1.0%, whereas none of 15 healthy volunteers or patients after allogeneic transplant had any detectable spontaneous IDO expression. The IDO expression increased by in vitro IFN-gamma stimulation in pregnant women (median 116%), healthy volunteers (median 11.7%) and patients with a low-grade aGvHD (grades 0-II) 28 days after transplant (median 433%) but not in patients with a severe aGvHD (grades III-IV) (median 0%), which was highly significant (P < 0.01). IDO expression was also measured in dendritic cells by qualitative RT-PCR, where a spontaneous IDO expression was detected in 16 of 31 (52%) pregnant women versus none of 17 healthy volunteers and none of 62 studied patients after transplant. IFN-gamma-induced IDO expression was detected in all pregnant women, all volunteers and 47 of 49 (96%) patients with a low-grade aGvHD (grades 0-II) after transplant, whereas only in two of 13 (16%) patients with aGvHD grade III IV was IFN-gamma-induced IDO expression observed. These data suggest that IDO expression might be involved in the development of allogeneic immune tolerance. PMID- 12588667 TI - The chemokines CCL5, CCL2 and CXCL12 play significant roles in the migration of Th1 cells into rheumatoid synovial tissue. AB - As the T-cell population in the synovial tissue (ST) in rheumatoid arthritis (RA) is dominated by T helper (Th) 1 cells, this study was designed to examine whether there is a preferential migration of polarized T cells to ST, and to identify the chemokines responsible for the migration. This was done by developing 10 T-cell clones specific for an arbitrary antigen (mouse immunoglobulin G (IgG)) from the peripheral blood (PB) of a healthy donor sensitized to mouse IgG. The Th polarizations of the clones were determined by measuring secreted interferon gamma and interleukin-4, following anti-CD3 stimulation. Migration to pools of RA ST cell-derived supernatants was analysed. Expression of the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CXCR3 and CXCR4 were analysed by flow cytometry. Th1 clones showed significantly higher migration to RA ST cell derived supernatant compared with Th2 clones. Blocking of either of the chemokines, CCL5 or CCL2, strongly inhibited migration of the Th1 cells between 56 and 77%, while blocking of CXCL12 inhibited migration between 44 and 61%. Blocking of CXCL10 had only a minor inhibitory effect. Our results demonstrate a selective migration of Th1 cells to RA ST supernatant and that blocking either CCL5, CCL2 or CXCL12 significantly inhibits T-cell migration. This indicates that CCL5, CCL2 and CXCL12 play significant roles in attracting Th1 cells towards the RA ST, and may prove potent targets for obstructing T-cell migration to the synovium. PMID- 12588671 TI - New aspects of metal allergy. AB - A common clinical manifestation of metal allergy is allergic contact dermatitis, however, there are some other exceptional manifestations such as lichen planus, pustulosis palmaris et plantaris including dyshidrotic eczema and severe form of alopecia areata. Evidence for such exceptional cases are: reproducible positive patch test reactions to various metal ions; analytical results of dental metal testing demonstrating the presence of metals to which the patients are undoubtedly hypersensitive; and dramatic curative effects of allergenic metal elimination. These dental metals are presumed to elute thus leading to the disease when they electrochemically turn to be a cathode. Cases which could not have been cured by antisymptomatic treatment but by allergen elimination are reported. PMID- 12588669 TI - Membrane androgen binding sites are preferentially expressed in human prostate carcinoma cells. AB - BACKGROUND: Prostate cancer is one of the most frequent malignancies in males. Nevertheless, to this moment, there is no specific routine diagnostic marker to be used in clinical practice. Recently, the identification of a membrane testosterone binding site involved in the remodeling of actin cytoskeleton structures and PSA secretion, on LNCaP human prostate cancer cells has been reported. We have investigated whether this membrane testosterone binding component could be of value for the identification of prostate cancer. METHODS: Using a non-internalizable testosterone-BSA-FITC analog, proven to bind on membrane sites only in LNCaP cells, we have investigated the expression of membrane testosterone binding sites in a series of prostate carcinomas (n = 14), morphologically normal epithelia, taken from areas of the surgical specimens far from the location of the carcinomas (n = 8) and benign prostate hyperplasia epithelia (n = 10). Isolated epithelial cells were studied by flow cytometry, and touching preparations, after 10-min incubation. In addition, routine histological slides were assayed by confocal laser microscopy. RESULTS: We show that membrane testosterone binding sites are preferentially expressed in prostate carcinoma cells, while BPH and non-malignant epithelial cells show a low or absent binding. CONCLUSIONS: Our results indicate that membrane testosterone receptors might be of use for the rapid routine identification of prostate cancer, representing a new diagnostic marker of the disease. PMID- 12588672 TI - Itching in patients suffering from psoriasis. AB - Some psoriatic patients suffer from intensive itching, however, literature data on its prevalence and especially on clinical manifestation are very limited. This study was undertaken to evaluate the frequency and clinical characteristics of itching in patients with psoriasis and to correlate the presence and intensity of pruritus with clinical severity of psoriasis. One hundred psoriatic individuals (psoriasis vulgaris in 77% and arthropatic psoriasis in 23%) were included in the study. The severity of psoriasis was assessed according to PASI score. Itching was evaluated using two methods: visual analog scale (VAS) and a specially designed questionnaire method. Itching was found in 80% of psoriatic patients. The severity of psoriasis in pruritic patients was significantly (p<0.004) higher as compared to non-pruritic subjects. Significant correlations were found between PASI scores and intensity of itching, as assessed by both scales: VAS and the questionnaire method (r=0.29, p<0.01 for both analyses). The presence and intensity of itching did not depend on age and gender of patients, type of psoriasis, duration of disease, and last outbreak of psoriasis. Generalized itching was reported by 28.7% of pruritic patients. The most common sites of itching were lower limbs (50%), trunk (48.7%), upper limbs (48.7%) and scalp (35%). Face appeared to be the least commonly affected skin area by itching (only 1.2%). We conclude that itching is a common symptom in patients with psoriasis, and its intensity correlates with clinical severity of the disease. PMID- 12588673 TI - Corrective dermatosurgery in childhood--own experience. AB - Congenital and acquired lesions on children's skin impose great burden upon the children and their parents. These children are frequently faced with various kinds of embarrassment in communication with their peers, ranging from sneering and mockery to isolation. That is why these lesions, especially if readily observable, should be completely removed or at least corrected as soon as possible to enable the child to establish normal communication with the environment and to prevent psychic trauma that may entail serious consequences later in life. Seven pediatric patients burdened with such lesions are presented (naevus papillomatous et verrucosus colli, atrophia linearis cutis faciei, naevus hyperkeratoticus systemicus dorsi, ichthyosis nigra, poikiloderma congenitum, hyalinosis cutis et mucosae, and alopecia post combustionem). Dermabrasion was used in six patients, whereas occipital hair graft transplantation to the alopecic sites was used in a girl with posttraumatic alopecia. PMID- 12588674 TI - Services for sexually transmitted infections in the new millennium--some thoughts from the United Kingdom. AB - Individuals who have acquired sexually transmitted infections (STIs) have been cared for, over time, by a wide variety of practitioners. In western Europe, most care for STIs is delivered in the primary care sector with dermatovereologists providing the specialist care. With a rising incidence of STIs across Europe, it is appropriate to look at different arrangements for the future. The developing plans in the United Kingdom (UK) are described with its national sexual health strategy, targets to be met, plans for greater use of nurses and primary care and possible future developments using the internet. PMID- 12588675 TI - Attitudes and behaviors of food donors and perceived needs and wants of food shelf clients. AB - OBJECTIVE: Explore the attitudes and behaviors of individuals who donate foods to food shelves and the perceived needs of the clientele using the food shelves in terms of cultural, health, and nutritional concerns. DESIGN: Qualitative methods (focus group interviews). SETTING: Focus groups (5 client and 7 donor) were conducted at food shelves, churches, and community centers. PARTICIPANTS: Client focus group members (n = 31) were mostly middle-aged and elderly individuals of mixed ethnicity and donor focus group members (n = 64) were mostly white and well educated and had incomes > $50 000. MAIN OUTCOME MEASURE(S): In-depth focus groups were conducted to obtain information regarding attitudes and beliefs of individuals who donate food and perceptions of needs among food shelf clients. ANALYSIS: Transcripts were analyzed systematically by coding comments according to categories and examining for common themes. RESULTS: Food donations did not match client needs for people with different ethnic backgrounds or age groups and food safety concerns. CONCLUSIONS AND IMPLICATIONS: This study demonstrates the need for nutrition educators to work with food shelf and food bank directors regarding the education of staff and general population on appropriate food donations. PMID- 12588676 TI - Personal attributes and job competencies needed by EFNEP paraprofessionals as perceived by EFNEP professionals. AB - OBJECTIVE: To determine the personal attributes and job competencies that are necessary for the job success of Expanded Food and Nutrition Education Program (EFNEP) paraprofessionals as perceived by EFNEP professionals. DESIGN: A qualitative descriptive design and a 3-round modified Delphi methodology was used. PARTICIPANTS: A convenience sample of 14 state and 20 county EFNEP professionals participated in all 3 Delphi rounds. The response rates for state and county professionals for all 3 Delphi rounds were 70% and 91%, respectively. VARIABLES MEASURED: Personal attributes and job competencies were generated and the level of importance was determined. ANALYSIS: An independent t test was employed to determine differences between importance scores by professional position. RESULTS: Thirty-seven personal attributes, 18 job competencies prior to hire, and 43 job competencies after training were generated and the majority were very important to important. County EFNEP professionals tended to score personal attributes and job competencies as more important than state EFNEP professionals. IMPLICATIONS: These personal attributes and job competencies prior to hire can guide hiring decisions and initial training content. The job competencies after training can shape content for inservice training. PMID- 12588677 TI - Associations between perceived family meal environment and parent intake of fruit, vegetables, and fat. AB - OBJECTIVE: To describe the family mealtime environment and assess associations with adult fruit, vegetable, and fat intake. DESIGN: Telephone survey. PARTICIPANTS: A convenience sample of 277 adults in the Minneapolis/St. Paul area were recruited through 4 schools. The sample was 85% female and 70% married. The mean number of children in the household was 2.6 (range 1 to 9). VARIABLES MEASURED: Adult fruit and vegetable intake, fat intake, and perceptions of the mealtime environment. ANALYSIS: Descriptive and mixed-model linear regression. RESULTS: Participants reported that the television was frequently on during dinner meals and almost one third felt that their family was too busy to eat dinner together. A higher frequency of television viewing during dinner was associated with lower fruit and vegetable consumption and higher fat consumption. Planning meals in advance was associated with higher fruit and vegetable consumption; however, 46% of the adults did not plan meals in advance. Arguments concerning eating behavior during dinner were associated with higher fat consumption. CONCLUSIONS AND IMPLICATIONS: The family meal environment is associated with adult eating patterns and should be considered when designing nutrition messages for families. PMID- 12588678 TI - Lessons learned from the "Spend Less. Eat Well. Feel Better." program efficacy trial. AB - "Spend Less. Eat Well. Feel Better." (SLEWFB) is a 3-hour intervention designed to improve food resource management ability, health, and food security status. This program efficacy trial targeted clients seeking financial aid to prevent homelessness. A randomized controlled design using a 1-hour food safety intervention as a control group was used. Four variables were targeted over a 30 day time period: (1) food security status, (2) basic financial management, (3) average fruit and vegetable intake, and (4) goal accomplishment. Repeated measures analysis of variance and chi-square analysis were used. Compared with control group participants (n = 47), SLEWFB graduates (n = 48) were less likely to have to choose between food and rent expenses (chi2 = 16.5, P =.00005) and improved their fruit and vegetable intake (F of time = 8.03, P <.006) regardless of financial assistance received. The findings imply that further investigation of coupling financial aid with effective life skills education is warranted. PMID- 12588679 TI - Community food security: background and future directions. AB - Nutrition educators research, teach, and conduct outreach within the field of community food security (CFS), yet no clear consensus exists concerning what the field encompasses. Nutrition education needs to be integrated into the CFS movement for the fundamental reason that optimal health, well-being, and sustainability are at the core of both nutrition education and CFS. Establishing commonalities at the intersection of academic research, public policy development, and distinctive nongovernmental organizations expands opportunities for professional participation. Entry points for nutrition educators' participation are provided, including efforts dedicated to education, research, policy, programs and projects, and human rights. PMID- 12588680 TI - Expression of potential lymphocyte trafficking mediator molecules in the mammary gland. AB - The mammary gland performs a variety of immunological functions, including protecting itself from mastitis and protecting neonates from infectious agents. Several molecules that mediate lymphocyte trafficking in the immune system are also expressed in the mammary gland. This review is focused on the immunological function of these molecules, especially glycosylation-dependent cell adhesion molecule-1 (GlyCAM-1) and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in the mammary gland. GlyCAM-1 is expressed in the lactating mouse mammary gland. Endothelial cells produce this protein and secrete it into milk. The glycosylated modification of mammary gland GlyCAM-1 is different from that of the lymph nodes, and lacks the binding ability for L-selectin on lymphocytes. GlyCAM-1 in the mammary gland is not involved in lymphocyte migration, and probably has another function besides that of the lymph nodes. MAdCAM-1 is expressed on endothelial cells of small venules around mouse mammary lobules during lactation. This molecule has the ability to interact with alpha4beta7 integrin on lymphocytes and mediates lymphocyte recruitment to the mammary gland. The density of beta7+/CD3+ T-cells is correlated with the density of the MAdCAM-1-stained area, suggesting that MAdCAM-1 may mediate the migration of these cells. In contrast, there is no relationship between MAdCAM-1 expression and the number of beta7+/c-IgA+ B-cells, implying that some other factor is involved in lymphocyte migration to the mammary gland. Chemokines, such as IL-8, GRO-alpha, MCP-1, RANTES and MEC, have been detected in human and mouse mammary glands. Although little information is available, these molecules may contribute to lymphocyte migration to the mammary gland. PMID- 12588681 TI - Metabolic changes in early lactation and impaired reproductive performance in dairy cows. AB - This review addresses the suggestion that the decline in dairy reproductive performance, as increasingly observed these days, may be due to a hampered process of metabolic adaptation in early lactating cows. In our opinion, adaptation to the negative energy balance is a gradual process. Because almost all cows do adapt in the long run, it is not possible to classify animals as adapted or non-adapted. The use of risk factors is more appropriate in this case and is discussed in this review. Among them are the body condition score and its derivatives, feed intake, the calculated negative energy balance, and metabolic parameters like the plasma concentration of insulin or the triacylglycerol content in the liver. Moreover, factors that play a role in the link between declined reproductive performance and the metabolic situation of the cow during the early lactating period are discussed. Among these are insulin, insulin-like growth factors, leptin, neuropeptide Y, non-esterified fatty acids, thyroid hormones, urea, and ammonia. PMID- 12588682 TI - The current status of major tick borne diseases in Zambia. AB - Tick-borne diseases occurring in Zambia are assuming more importance as they continue to be a major economic problem not only in Zambia, but in many parts of Eastern, Southern and Central Africa. The current control methods, which include the use of toxic acaricides to kill ticks, and the virulent sporozoite infection and treatment method have limitations. Recombinant vaccines, currently in their experimental stages, offer hope for the future. The use of acaricides is hampered by the development of acaricide resistance and live vaccines are dependent on cold chain facilities, which are a formidable obstacle in the poorly developed infrastructure in parts of Zambia where the vaccine is most needed. Amidst these drawbacks are the results of the recent research on parasites and vector recombinant vaccines which promise to circumvent these problems. The history, current status and attitudes regarding the control of these diseases, taking into account their complexity, are reviewed. The establishment of the well-designed Central Veterinary Research Institute (CVRI) and Japanese International Cooperation Agency (JICA) sponsored veterinary school, both have a potential for high quality research, with access to a wealth of specimens a veritable goldmine of research material. It is thus hoped that this review will stimulate the desire to maximize the value of the tick and tick-borne disease research in both Zambia and the international research community. PMID- 12588683 TI - Left displacement of the abomasum in dairy cattle: recent developments in epidemiological and etiological aspects. AB - The research with respect to displacement of the abomasum (DA) in dairy cattle is reviewed. Evaluated articles describe epidemiological and experimental studies. The occurrence is elevated with regard to breed, gender, age, concurrent diseases, environmental aspects and production levels as contributing factors and emphasis is placed on the effects of nutrition and metabolism. Reviewing the experimental work, distinction is made between the research into gas production in the abomasum and hypomotility of the abomasum, since both represent presumed pathways in the development of DA. Although the different fields of research have positive contributions to the understanding of the pathogenesis of DA, contradictions in the different studies are present. This is partly due to extrapolation of results from sheep to cows, or because of a low number of cows in the experiments. Finally, general suggestions are made for further research in the field of the pathogenesis of DA. PMID- 12588684 TI - Analysis of selective mobilization of L-selectin and Mac-1 reservoirs in bovine neutrophils and eosinophils. AB - Following activation of granulocytes, L-selectin (CD62L) is generally shed from the cellular surface, whereas Mac-1 (CD11b/CD18) expression is well known to increase. However, a number of studies in bovines and humans show that the expression of L-selectin may increase as well. This urged us to examine the possible existence of both L-selectin and Mac-1 reservoirs in bovine neutrophil and eosinophil populations through the use of flow cytometry in combination with an optimized method for cell membrane permeabilization. Augmented L-selectin and Mac-1 expression was detected in both granulocyte populations upon saponin treatment. Confocal microscopic studies indicated that both molecules exhibit a different pattern of subcellular localization. Incubation with sialidase revealed the existence of hidden L-selectin epitopes at the cell surface, while no additional Mac-1 epitopes were exposed. Platelet-activating factor stimulation decreased surface and total expression of L-selectin to the same extent in both populations, but solely affected Mac-1 surface expression on eosinophils. Moreover, cytoskeletal actin filaments and microtubules were found to be involved in the regulation of Mac-1 surface expression on bovine neutrophils and eosinophils. In marked contrast, expression of L-selectin was minimally affected by cytoskeleton perturbing agents. The present study indicates that L-selectin and Mac-1 adhesion molecules reside in distinctly located reservoirs in bovine granulocytes and can be selectively mobilized upon in vitro stimulation. PMID- 12588685 TI - Standardisation and comparison of serial dilution and single dilution enzyme linked immunosorbent assay (ELISA) using different antigenic preparations of the Babesia (Theileria) equi parasite. AB - Serial dilution and single dilution enzyme linked immunosorbent assays (ELISA) were standardised and their sensitivity and specificity were compared for serodiagnosis of Babesia equi infection. The antibody titres of 24 donkey sera of known identity were determined separately by serial dilution ELISA using three different B. equi antigens namely whole merozoite (WM), cell membrane (CM) and high speed supernatant (HSS). The ratios of the optical density (OD) of known positive and known negative sera at different serum dilutions were calculated and termed as the positive/negative (P/N) ratio. The coefficients of correlation (r) were calculated between the P/N ratios at different dilutions of sera and the log10 antibody titres of the same sera were ascertained by serial dilution ELISA. The highest value of 'r' was obtained at a serum dilution of 1:200. From log10 antibody titre of sera (y) and their P/N ratio at a dilution of 1:200 (x), regression equations (y = a + bx) were calculated separately for the three antigens. Test sera were diluted to 1:200, their OD were read in duplicate wells and were converted to the P/N ratio. Antibody titres were predicted from the P/N ratio using a regression equation separately for the three antigens. Titres obtained by both ELISAs were not significantly different from each other, thus confirming that single dilution ELISA could be successfully used to replace conventional serial dilution ELISA. The sensitivity, specificity and predictive value of single dilution ELISA was validated statistically using 42 B. equi disease-positive sera and 106 B. equi disease-negative sera. The WM antigen was found to be the most sensitive with a higher predictive value for negative test sera as compared to the CM or HSS antigens. Sera positive for other equine infections including Babesia caballi showed no cross-reaction with the three B. equi antigens in ELISA, thus the test was immunologically specific. Antibody titres of 109 unknown field donkey/horse sera obtained by serial and single dilution ELISA using the WM antigen did not show any significant difference. Since the single dilution ELISA was found to be more economical, convenient, sensitive, specific than the serial dilution ELISA and has a high predictive value, it is suitable for use in sero-epidemiological studies on B. equi infections in the field. PMID- 12588686 TI - An individual modelling tool for within and between lactation consecutive cases of clinical mastitis in the dairy cow: an approach based on a survival model. AB - Clinical mastitis in dairy cows has for many years been the subject of numerous epidemiological surveys to determine the main risk factors. In most cases this data has been analysed using a standard Poisson model without taking into consideration possible dependence between consecutive pathological events. These analyses have brought to light a great many potential risk factors without making it possible to clarify a certain amount of confusion surrounding the effects. The extension of an individual within a lactation model, considering dependence between clinical cases of mastitis within lactation so as to take into account inter-lactation dependence (which has already been published) is presented in the form of mixed distributions within the same survival model framework. By introducing new parameters, infection rate at calving and the identification of a higher exogenous infection rate indoors than at pasture, it is possible to take into consideration what had previously appeared to be a lactation stage factor, a calving month factor or even part of a parity factor. By considering these two types of dependence within the same model, it appears to be possible to obtain a simpler model in terms of the factors to be taken into account, and one that is based on generally acknowledged and easily understandable biological considerations. Lastly, a possible way of extending the model is to consider the dry period before calving and this is presented. This would make it possible to envisage developing a complete model of the animal's lifetime in the not-too distant future. It is still necessary, however, to determine the farming system factors in the general sense of the term, which specifically affect one or the other of the different model parameters, before one can draw conclusions as to the potential extension of this type of model. A national survey is currently being carried out on approximately 600 French breeding farms that will help meet this last objective. PMID- 12588687 TI - Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis. AB - A reverse transcription-PCR (RT-PCR) method is presented for the highly sensitive and specific detection of foot-and-mouth disease virus (FMDV). A primer pair flanking a region of the viral polymerase gene (3D) corresponding to the C terminus of the protein was designed and a single step RT-PCR reaction was developed. The assay allowed the detection of viral RNA from a variety of animal samples and from a wide range of FMDV isolates of different origins and serotypes. The presence of an Ahd I restriction site within the amplicon in 96% of the isolates analyzed allowed an additional confirmation step of the positive reactions by a simple digestion yielding characteristic fragment sizes. The set of primers described here was suitable for direct sequencing of the PCR product (290 bp), and the nucleotide sequences corresponding to the SAT 1 and SAT 3 strains were determined. The segment amplified, when used in phylogenetic studies, allowed the clustering of SAT isolates and the rest of FMDV strains as two separate lineages. PMID- 12588688 TI - Proteic boost enhances humoral response induced by DNA vaccination with the dnaK gene of Chlamydophila abortus but fails to protect pregnant mice against a virulence challenge. AB - In order to enhance the quantity and the protective properties of the antibodies induced by DNA vaccination with the heat shock protein dnaK gene of Chlamydophila abortus AB7 as well as to elicit an efficient cellular immune response, we vaccinated mice with a DNA prime followed by a boost with the recombinant DnaK protein. In non-pregnant mice, this strategy induced the same predominance of the IgG2a isotype as DNA immunization alone with a substantial increased antibody level. The induced antibodies had no in vitro neutralizing properties on C. abortus infectivity. Moreover, the proteic boost probably failed to elicit an efficient cellular immune response since the pregnant or non-pregnant mice were not protected against the bacterial challenge. PMID- 12588689 TI - Virulence stability in Flavobacterium psychrophilum after storage and preservation according to different procedures. AB - Experimental infections and lethal dose 50% (LD 50) evaluation were conducted in rainbow trout fingerlings, using a virulent strain of Flavobacterium psychrophilum processed and stored or maintained in different ways; lyophilisation, freezing at -80 degrees C, maintenance in enriched Anacker and Ordal (EAO) medium at 4 degrees C, revival and subsequent in vivo passages in fish. Experiments were performed 1, 8 and 23 months after storing the bacteria. Out of a total of 12 cultures revived for experimentation, one failed to grow and another was found to express modified properties including decreased virulence in spite of in vivo passages. In all other cases, whatever the conditions of preservation, virulence was fairly well maintained after 1 and 8 months of storage. In the last test, after 23 months, the bacteria maintained in the EAO medium at 4 degrees C were found significantly attenuated. Conversely, lyophilised and frozen bacteria only expressed a slight increase in LD0. It was concluded that virulent strains of F. psychrophilum were likely to retain their properties without special provisions within limited periods of time, and that both lyophilisation and freezing at -80 degrees C were reliable methods for long term preservation of virulence. PMID- 12588690 TI - Eating frequency and the risk of colon cancer. AB - Eating frequency has been found in most previous studies to have a positive association or no association with colon cancer. We report data from a large case control study to determine the effect of eating frequency on colon cancer risk. Data were analyzed from interviews of 1,966 cases of colon cancer and 2,380 controls from selected areas in Northern California, Utah, and Minnesota. Respondents were asked whether they usually ate or drank something besides water at eight different occasions during the day. We controlled for age, family history of colorectal cancer, body mass index, physical activity, intake of non steroidal anti-inflammatory medication, and dietary intake of energy, fiber, and calcium. In fully adjusted models, we found no significant associations between number of daily eating occasions and colon cancer in women. In men, risk of overall colon cancer was lower for one to two times per day (odds ratio = 0.54, 95% confidence interval = 0.36-0.83) than for three times per day, but risk was not increased for more than three times per day. Compared with three times per day, we found no evidence for an association between colon cancer risk and eating frequencies more than three times per day. The increased risk of colon cancer limited to men eating less than three times per day may be due to uncontrolled confounding. PMID- 12588692 TI - Diet quality and cancer incidence in Nova Scotia, Canada. AB - Cancer rates in the province of Nova Scotia, Canada, are among the highest in the country and coincide with elevated rates of risk factors such as smoking, poor diet, and obesity. To investigate the importance of diet on cancer, using data from the 1990 Nova Scotia Nutrition Survey, we developed a diet quality score reflecting compliance with 17 nutrient recommendations. The survey data were subsequently linked with the provincial cancer registry, and the relationship between diet quality and cancer was quantified using logistic regression. Our results support an inverse relationship between diet quality and cancer, although limited statistical power resulting from our small study sample did not reveal any statistically significant relationships. We estimated that cancer incidence could potentially be reduced by approximately 35% through improved diet quality. On the basis of poor diet, nutrition-related factors (high body mass index), our estimates of the preventable fraction of cancer, and the high provincial cancer rates, we recommend health promotion strategies aimed at improving diet quality in Nova Scotia. PMID- 12588693 TI - Dietary and reproductive factors associated with benign breast disease in Mexican women. AB - Benign breast disease (BBD) is common in women in the reproductive age group. The results of epidemiological studies that have examined the relationship between diet and BBD are controversial and scarce. The aim of the present study was to identify and evaluate the impact of dietary and reproductive risk factors in the development of BBD in Mexican women. Between 1994 and 1996, 121 women with BBD and 121 age-matched (+/- 3 yr) clinical controls with non-breast-related diseases were identified. The study population was directly interviewed about their reproductive history, and a semiquantitative food frequency questionnaire was used to obtain dietary information. The population was originally identified as part of a hospital-based case-control study examining exposure to 1,1-bis(p chlorophenyl)-2,2,2-trichloroethane and breast cancer. A significant reduction in the risk of BBD was observed when comparing the upper with the lower tertiles of consumption of citrus fruit [odds ratio (OR) = 0.43, 95% confidence interval (CI) = 0.21-0.88], non-citrus fruit (OR = 0.35, 95% CI = 0.16-0.76), diary products (OR = 0.36, 95% CI = 0.18-0.75), and food sources of lignans (OR = 0.48, 95% CI = 0.24-0.98). A marginally significant reduction in the risk of BBD associated with the consumption of vitamin B-12 (OR = 0.48, 95% CI = 0.23-1.02) was also observed. Our results add new information about the role of diet in the etiology of BBD. PMID- 12588694 TI - Methods to increase fruit and vegetable intake with and without a decrease in fat intake: compliance and effects on body weight in the nutrition and breast health study. AB - Dietary patterns that involve a decrease in fat and an increase in fruit and vegetable (FV) intake have been suggested to decrease cancer risks. In this study, intervention methods to selectively modify dietary fat and/or FV intakes were developed. Compliance to the diets and the effects on body weight are shown, because both of these dietary changes can impact on and be confounded by changes in energy intake. A total of 122 women with a family history of breast cancer were randomized onto one of four diets for 12 mo. Counseling methods were devised to increase amount and variety of FV consumed with or without a decrease in fat intake using modified exchange list diets. Women on the low-fat and combination low-fat/high-FV diet arms decreased their fat intakes to approximately 16% of energy. Women on the high-FV and the combination low-fat/high-FV diet arms increased FV intakes to approximately 11 servings/day. Despite counseling efforts to maintain baseline energy intakes, mean body weight increased significantly by 6 pounds in women in the high-FV diet arm and decreased significantly by 5 pounds in women in the low-fat diet arm. Percent body fat also was increased in the high FV diet arm and decreased in the low-fat diet arm. Body weight and percent body fat in the combination diet arm did not change significantly. Control of energy intake, therefore, appears to have been achieved only when the addition of FV to the diet was balanced by a decrease in fat intake and both dietary components were enumerated daily. Maintenance of energy intake, therefore, did not appear to be attained intrinsically when individuals were counseled to make changes in the composition of their diets. PMID- 12588695 TI - Relationship of folate, vitamin B-6, vitamin B-12, and methionine intake to incidence of colorectal cancers. AB - It is hypothesized that diets deficient in folate, methionine, and vitamins B-6 and B-12 cause DNA hypomethylation and, as a result, increase risk of colorectal cancers. Furthermore, it is proposed that alcohol, a methyl group antagonist, increases risk of colorectal cancers among those with low intake of folate. Data from the Iowa Women's Health Study, a population-based cohort of incident cancer, were used to examine the relationship of folate, methionine, and vitamins B-6 and B-12 to occurrence of cancers of the colon (n = 598) and rectum (n = 123) over 13 yr of follow-up. There were no independent associations of folate, methionine, or vitamins B-6 and B-12 derived from a food frequency questionnaire with incidence of colon cancer. Adjusted relative risks (RRs) of rectal cancer were similar across categories of folate, vitamin B-12, and methionine intake, but RRs increased progressively with increasing intake of vitamin B-6 [P (for trend) = 0.03]. RRs suggested that incidence of cancer of the proximal colon was lower among those with 1) high folate and high vitamin B-12 intake [RR = 0.59, 95% confidence interval (CI) = 0.39-0.89] and 2) high folate and high vitamin B-6 intake (RR = 0.65, 95% CI = 0.50-0.84) than among those with the lowest intake of these nutrients. Incidence of cancer of the proximal colon was also somewhat lower among those with high folate and low alcohol intake (RR = 0.44, 95% CI = 0.22-0.89). Findings provide limited support for an association between dietary factors involved in DNA methylation and risk of cancers of the colon and rectum. PMID- 12588696 TI - Breast cancer, serum antioxidant vitamins, and p53 protein overexpression. AB - Although several studies have suggested that the relationship between the various risk factors and breast cancer may differ according to the breast cancer subtypes, it is not clear whether the p53 protein expression status of breast cancer represents an etiologically distinct form of the disease with different risk factor profiles. This study was carried out to investigate whether the relationship between breast cancer and serum antioxidant vitamins would differ according to p53 expression status (p53-positive and p53-negative). Breast cancer cases (n = 92) and controls (n = 122) were recruited between January 1993 and April 1994 at the Asan Medical Center. p53 overexpression in tissue sections from 92 women with breast cancer was determined using immunohistochemistry. The serum concentrations of the carotenoids and alpha-tocopherol were measured using liquid chromatography. For serum antioxidant vitamins, odds ratios with respect to the common control group for breast cancers of different p53 protein overexpression status were compared using multiple polytomous logistic regression. Serum concentrations of beta-carotene and zeaxanthin + lutein were significantly associated with the risk of breast cancer in p53-positive and p53-negative cancers. The adjusted odds ratios for the highest compared with the lowest quartile were 0.19 [95% confidence interval (CI) = 0.01-2.51] for beta-carotene and 0.34 (95% CI = 0.06-1.87) for zeaxanthin + lutein in p53-positive cancer cases and 0.05 (95% CI = 0.00-0.57) for beta-carotene and 0.06 (95% CI = 0.01 0.40) for zeaxanthin + lutein in p53-negative cancer cases. However, none of these associations differed significantly between p53-positive and p53-negative cancers. The results of this study showed that the relationship of antioxidant vitamins with breast cancer does not differ according to the presence or absence of the p53 mutation. PMID- 12588697 TI - Detection of Bowman-Birk inhibitor and anti-Bowman-Birk inhibitor antibodies in sera of humans and animals treated with Bowman-Birk inhibitor concentrate. AB - The Bowman-Birk inhibitor (BBI) is a soybean-derived serine protease inhibitor with anticarcinogenic activities. BBI, in the form of BBI concentrate (BBIC), is currently being evaluated in clinical trials as a human cancer-preventive agent. In the present study, an enzyme-linked immunosorbent assay was used to measure BBI concentrations in serum samples collected from human subjects and animals treated with BBIC. The results demonstrate that the serum BBI concentration was higher than the baseline level for the patients after treatment with BBIC at 100 800 chymotrypsin-inhibitor units/day for 0.5, 1, 2, 4, and 6 mo. The increase in serum BBI concentration was also observed in dogs treated with BBIC at 100-1,000 mg/kg/day for 52 wk, and the increase was dose dependent. The results also indicate that anti-BBI antibodies were present in animals and the serum levels of anti-BBI antibodies increased significantly in mice treated with BBIC at 100 1,000 mg/kg/day for 15 and 26 wk. The increase in the serum level of anti-BBI antibodies in dogs treated with BBIC was not statistically significant, and no increase in the serum level of anti-BBI antibodies was observed in human subjects after BBIC treatment. These results suggest that orally ingested BBI is absorbed by human subjects and animals and that some animals develop antibodies to BBI in response to treatment with BBIC. PMID- 12588698 TI - Effects of Bowman-Birk inhibitor on rat colon carcinogenesis. AB - The present study was undertaken to determine whether the Bowman-Birk inhibitor (BBI) could prevent colon carcinogenesis in rats treated with dimethylhydrazine (DMH) and whether there were adverse side effects associated with treatment with BBI for cancer prevention. BBI was evaluated in the forms of purified BBI (PBBI) or an extract of soybeans enriched in BBI, termed BBI concentrate (BBIC). The results demonstrate that PBBI and BBIC reduced the incidence and frequency of tumors in DMH-treated rats compared with animals treated with DMH alone. Autoclaved BBIC, in which the protease inhibitor activity of BBI was destroyed, had a weak and statistically insignificant, suppressive effect on DMH-induced colon carcinogenesis in rats, suggesting that the protease inhibitor activity of BBI is likely to be responsible for the anticarcinogenic activity of BBIC. Soy molasses, which contains soy isoflavones, did not have an effect on colon cancer carcinogenesis in DMH-treated rats. Similar to results from previous studies (Nauss et al. JNCI 73, 915-924, 1984), the most aggressive, malignant colon adenocarcinomas developed within or in association with gut-associated lymphoid tissue aggregates. No adverse side effects on the pancreas or animal growth were observed in rats treated with PBBI or BBIC. These results demonstrate that PBBI and BBIC may be used to prevent colon cancer without significant adverse side effects. PMID- 12588699 TI - Dietary flaxseed inhibits human breast cancer growth and metastasis and downregulates expression of insulin-like growth factor and epidermal growth factor receptor. AB - Recent studies indicate that diets rich in phytoestrogens and n-3 fatty acid have anticancer potential. This study determined the effect of flaxseed (FS), the richest source of lignans and alpha-linolenic acid, on growth and metastasis of established human breast cancer in a nude mice model. Estrogen receptor-negative human breast cancer cells, MDA-MB-435, were injected into the mammary fat pad of mice (Ncr nu/nu) fed a basal diet (BD). At Week 8, mice were randomized into two diet groups, such that the groups had similar tumor size and body weight. One continued on the BD, while the other was changed to BD supplemented with 10% FS, until sacrifice at Week 15. A significant reduction (P < 0.05) in tumor growth rate and a 45% reduction (P = 0.08) in total incidence of metastasis were observed in the FS group. Lung metastasis incidence was 55.6% in the BD group and 22.2% in the FS group, while the lymph node metastasis incidence was 88.9% in the BD group and 33.3% in the FS group (P < 0.05). Mean tumor number (tumor load) of total and lymph node metastasis was significantly lower in the FS than in the BD group (P < 0.05). Metastatic lung tumor number was reduced by 82%, and a significantly lower tumor trend (P < 0.01) was observed in the FS group. Lung weight, which also reflects metastatic tumor load, in the FS group was reduced by 20% (P < 0.05) compared with the BD group. Immunohistochemical study showed that Ki-67 labeling index and expression of insulin-like growth factor I and epithelial growth factor receptor in the primary tumor were lower in the FS (P < 0.05) than in the BD group. In conclusion, flaxseed inhibited the established human breast cancer growth and metastasis in a nude mice model, and this effect is partly due to its downregulation of insulin-like growth factor I and epidermal growth factor receptor expression. PMID- 12588700 TI - Increased cellular carotenoid levels reduce the persistence of DNA single-strand breaks after oxidative challenge. AB - Dietary antioxidants, such as the carotenoids, may protect DNA from oxidative damage. This has been proposed to explain the epidemiological association between higher consumption of fruits and vegetables, which are rich in antioxidants, and lower incidence of cancer. However, this remains to be demonstrated conclusively. The effects of carotenoid supplementation on 1) baseline DNA damage, 2) susceptibility of cellular DNA to oxidative attack, and 3) DNA repair were measured in the human lymphocyte cell line Molt-17. Baseline DNA damage, susceptibility to oxidant attack (100 mumol/l H2O2 for 5 min at 4 degrees C), and disappearance of DNA single-strand breaks (SSB) after oxidative challenge were monitored by single-cell gel electrophoresis. DNA repair patch synthesis activity in cell extracts was determined using assays that measure nucleotide incorporation during repair of oxidative lesions in template DNA. Unlike single cell gel electrophoresis, the parameters measured with these assays are not dependent on strand break religation. There was no evidence that beta-carotene, lutein, or beta-cryptoxanthin supplementation protected cellular DNA from oxidation under basal conditions or after oxidative challenge. However, only carotenoid-supplemented cells exhibited a significant decrease in numbers of SSB over a 2-h period after treatment with H2O2. Carotenoid supplementation did not provoke any detectable change in repair patch synthesis activity. We conclude that supplementation with carotenoids at 8 mumol/l does not provide significant antioxidant protection for DNA in Molt-17 lymphocytes but may enhance recovery of cells from oxidative challenge, as measured by loss of SSB. We argue that these data are most consistent with carotenoids acting to enhance DNA strand break repair. PMID- 12588701 TI - Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by fermented soy milk. AB - The effect of a fermented soy milk product (FSP) on various human breast carcinoma cell lines was investigated, and it was shown to have a growth inhibitory effect, especially on MCF-7 cells. Thus the MCF-7 cell line was used to study the mechanism of action. In female severe combined immune deficiency mice implanted with MCF-7 cells, pretreatment with FSP significantly inhibited tumor growth. The inhibitory effect of FSP on MCF-7 cells seemed to be caused by the additive effects of a wide variety of constituents. The active components of FSP are mainly in the water phase, and the lipid-soluble fraction, which includes the soy isoflavones such as genistein and daidzein, is relatively ineffective. A variety of methods were used to demonstrate that FSP caused apoptotic cell death in MCF-7 cells. FSP induced generation of reactive oxygen species (ROS). Growth inhibition and ROS generation induced by FSP could be inhibited by catalase and deferoxamine, indicating that the ROS production probably was the cause of this apoptotic cell death. This study suggests that FSP retards tumor growth in vivo and can trigger apoptosis in vitro. It may, therefore, be a potential nutritional supplement in chemotherapy. PMID- 12588702 TI - RRR-alpha-tocopheryl succinate induction of DNA synthesis arrest of human MDA-MB 435 cells involves TGF-beta-independent activation of p21Waf1/Cip1. AB - RRR-alpha-tocopheryl succinate (vitamin E succinate, VES), a derivative of vitamin E, is a potent antitumor agent. Cellular events involved in VES-induced DNA synthesis arrest of human MDA-MB-435 breast cancer cells were studied. VES induces a dose- and time-dependent inhibition of DNA synthesis and a G0/G1 cell cycle arrest. VES induces expression of p21Waf1/Cip1 mRNA and protein, and antisense oligomers to p21 block VES-induced growth arrest. Evidence suggesting that VES modulates p21 expression in a transforming growth factor-beta (TGF-beta) independent fashion includes failure of TGF-beta-neutralizing antibodies to block VES-induced DNA synthesis arrest or VES activation of a p21 promoter-regulated reporter gene; VES is not capable of inducing the translocation of green fluorescent protein-Smad2 into the nucleus and is not capable of stimulating a TGF-beta-dependent reporter gene, and VES induces growth inhibition and upregulates p21 mRNA levels in TGF-beta receptor-defective cells. PMID- 12588703 TI - Interleukin-9 influences chemokine release in airway smooth muscle: role of ERK. AB - Interleukin (IL)-9 is a pleiotropic cytokine that has been proposed as a candidate gene for asthma. As IL-9 expression is correlated with airway hyperresponsiveness in animals, we examined the effects of IL-9 on cultured human airway smooth muscle (HASM) cells. IL-9 alone had no effect on IL-8 release, but at concentrations of > or =30 ng/ml, IL-9 significantly increased IL-8 release induced by TNF-alpha. IL-9 increased phosphorylation of extracellular signal regulated protein kinase (ERK, p42 and p44) in a concentration- and time dependent fashion, and U-0126 (10 micro M), which inhibits ERK phosphorylation, abolished the synergism between TNF-alpha and IL-9 on IL-8 release. IL-9 alone had no effect on eotaxin release into HASM cell supernatants but at concentrations of > or =10 ng/ml caused an approximately 50% increase in release of eotaxin evoked by IL-13 (10 ng/ml). U-0126 blocked the synergism between IL-9 and IL-13 on eotaxin release. IL-9 had no effect on cyclooxygenase-2 (COX-2) expression or PGE(2) release and did not augment the COX-2 expression that was induced by IL-1beta. Our results indicate that airway smooth muscle is a target for IL-9 and that IL-9 amplifies the potential for these cells to recruit eosinophils and neutrophils into the airways by a mechanism involving ERK. PMID- 12588705 TI - IFN-gamma inhibits human airway smooth muscle cell proliferation by modulating the E2F-1/Rb pathway. AB - Elucidating the factors that inhibit the increase in airway smooth muscle (ASM) mass may be of therapeutic benefit in asthma. Here, we investigated whether interferon-gamma (IFN-gamma), a potent inducer of growth arrest in various cell types, regulates mitogen-induced ASM cell proliferation. IFN-gamma (1-100 U/ml) was found to markedly decrease both DNA synthesis and ASM cell number induced by the mitogens epidermal growth factor (EGF) and thrombin. Interestingly, IFN-gamma had no effect on mitogen-induced activation of three major mitogenic signaling pathways, phosphatidylinositol 3-kinase, p70(S6k), or mitogen-activated protein kinases. Mitogen-induced expression of cell cycle regulator cyclin D1 was increased by IFN-gamma, whereas no effect was observed on degradation of p27(Kip1). Expression array analysis of 23 cell cycle-related genes showed that IFN-gamma inhibited EGF-induced increases in E2F-1 expression, whereas induction of c-myc, cyclin D2, Egr-1, and mdm2 were unaffected. Induction of E2F-1 protein and Rb hyperphosphorylation after mitogen stimulation was also suppressed by IFN gamma. In addition, IFN-gamma decreased activation of cdk2 and expression of cyclin E, upstream signaling molecules responsible for Rb hyperphosphorylation in the late G1 phase. IFN-gamma also increased levels of IFI 16 protein, whose mouse homolog p202 has been associated with growth inhibition. Together, our data indicate that IFN-gamma is an effective inhibitor of ASM cell proliferation by blocking transition from G1-to-S phase by acting at two different levels: modulation of cdk2/cyclin E activation and inhibition of E2F-1 gene expression. PMID- 12588704 TI - Mucin secretion and PKC isoforms in SPOC1 goblet cells: differential activation by purinergic agonist and PMA. AB - SPOC1 cells, which are a mucin-secreting model of rat airway goblet cells, possess a luminal P2Y2 purinoceptor through which UTP, ATP, and ATPgammaS stimulate secretion with EC50 values of approximately 3 microM. PMA elicits mucin secretion with high EC50 (75 nM) and saturation (300 nM) values. For the first time in airway mucin-secreting cells, the PKC isoforms expressed and activated by a secretagogue were determined using RT-PCR/restriction-enzyme mapping and Western blotting. Five isoforms were expressed: cPKCalpha, nPKCdelta and -eta, and aPKCzeta and -iota/lambda. PMA caused cPKCalpha and nPKCdelta to translocate to the membrane fraction of SPOC1 cells; only nPKCdelta so responded to ATPgammaS. Membrane-associated nPKCdelta and mucin secretion increased in parallel with ATPgammaS concentration and yielded EC50 values of 2-3 microM and maximum values of 100 microM. Effects of PMA to increase membrane-associated cPKCalpha and nPKCdelta saturated at 30 nM, whereas mucin secretion saturated at 300 nM, which suggests a significant PKC-independent effect of PMA on mucin secretion. A prime alternate phorbol ester-receptor candidate is the C1-domain protein MUNC13. RT-PCR revealed the expression of ubiquitous (ub)MUNC13-2 and its binding partner, DOC2-gamma. Hence, P2Y2 agonists activate nPKCdelta in SPOC1 cells. PMA activates cPKCalpha and nPKCdelta at high affinity and stimulates a lower affinity PKC-independent pathway that leads to mucin secretion. PMID- 12588706 TI - Antimacrophage chemokine treatment prevents neutrophil and macrophage influx in hyperoxia-exposed newborn rat lung. AB - Macrophage-derived cytokines may provoke the inflammatory response in lung injury. Because macrophage influx is a prominent feature of the cellular inflammatory response accompanying the development of bronchopulmonary dysplasia, we hypothesized that blocking macrophage influx would reduce overall cellular influx and oxidative damage. Newborn rats were exposed at birth to 95% O(2) or air for 1 wk, and hyperoxia-exposed pups were injected with anti-monocyte chemoattractant protein-1 (MCP-1) or IgG control on days 3-5. MCP-1 was increased in bronchoalveolar lavage fluid and in histological sections from the 95% O(2) exposed, IgG-injected pups compared with air-exposed controls. At 1 wk, anti-MCP 1-treated pups had reduced leukocyte numbers, both macrophages and neutrophils, in bronchoalveolar lavage fluid compared with IgG-treated controls. Cytokine induced neutrophil chemoattractant-1, the rat analog of IL-8, was not significantly decreased in lavage fluid but was reduced in lung cells in anti-MCP 1-treated pups. Tissue carbonyls, a measure of protein oxidation, were decreased in anti-MCP-1-treated pups. Anti-MCP-1 treatment prevented neutrophil influx and reduced protein oxidation in hyperoxia-exposed newborn rats. PMID- 12588707 TI - Mild hypoxia impairs alveolarization in the endothelial nitric oxide synthase deficient mouse. AB - In addition to its vasodilator properties, nitric oxide (NO) promotes angiogenesis in the systemic circulation and tumors. However, the role of NO in promoting normal lung vascular growth and its impact on alveolarization during development or in response to perinatal stress is unknown. We hypothesized that NO modulates lung vascular and alveolar growth and that decreased NO production impairs distal lung growth in response to mild hypoxia. Litters of 1-day-old mouse pups from parents that were heterozygous for endothelial nitric oxide synthase (eNOS) deficiency were placed in a hypobaric chamber at a simulated altitude of 12,300 ft (Fi(O(2)) = 0.16). After 10 days, the mice were killed, and lungs were fixed for morphometric and molecular analysis. Compared with wild-type controls, mean linear intercept (MLI), which is inversely proportional to alveolar surface area, was increased in the eNOS-deficient (eNOS -/-) mice [51 +/ 2 micro m (eNOS -/-) vs. 41 +/- 1 micro m (wild type); P < 0.01]. MLI was also increased in the eNOS heterozygote (+/-) mice (44 +/- 1 micro m; P < 0.03 vs. wild type). Vascular volume density was decreased in the eNOS -/- mice compared with wild-type controls (P < 0.03). Lung vascular endothelial growth factor (VEGF) protein and VEGF receptor-1 (VEGFR-1) protein content were not different between the study groups. In contrast, lung VEGFR-2 protein content was decreased from control values by 63 and 34% in the eNOS -/- and eNOS +/- mice, respectively (P < 0.03). We conclude that exposure to mild hypoxia during a critical period of lung development impairs alveolarization and reduces vessel density in the eNOS deficient mouse. We speculate that NO preserves normal distal lung growth during hypoxic stress, perhaps through preservation of VEGFR-2 signaling. PMID- 12588708 TI - PKA inhibits RhoA activation: a protection mechanism against endothelial barrier dysfunction. AB - Much evidence indicates that cAMP-dependent protein kinase (PKA) prevents increased endothelial permeability induced by inflammatory mediators. We investigated the hypothesis that PKA inhibits Rho GTPases, which are regulator proteins believed to mediate endothelial barrier dysfunction. Stimulation of human microvascular endothelial cells (HMEC) with thrombin (10 nM) increased activated RhoA (RhoA-GTP) within 1 min, which remained elevated approximately fourfold over control for 15 min. The activation was accompanied by RhoA translocation to the cell membrane. However, thrombin did not activate Cdc42 or Rac1 within similar time points, indicating selectivity of activation responses by Rho GTPases. Pretreatment of HMEC with 10 micro M forskolin plus 1 micro M IBMX (FI) to elevate intracellular cAMP levels inhibited both thrombin-induced RhoA activation and translocation responses. FI additionally inhibited thrombin mediated dissociation of RhoA from guanine nucleotide dissociation inhibitor (GDI) and enhanced in vivo incorporation of (32)P by GDI. HMEC pretreated in parallel with FI showed >50% reduction in time for the thrombin-mediated resistance drop to return to near baseline and inhibition of approximately 23% of the extent of resistance drop. Infection of HMEC with replication-deficient adenovirus containing the protein kinase A inhibitor gene (PKA inhibitor) blocked both the FI-mediated protective effects on RhoA activation and resistance changes. In conclusion, the results provide evidence that PKA inhibited RhoA activation in endothelial cells, supporting a signaling mechanism of protection against vascular endothelial barrier dysfunction. PMID- 12588709 TI - A new role for glucocorticoids in septic shock: balancing the immune response. PMID- 12588710 TI - Immunity to nontypeable Haemophilus influenzae: elucidating protective responses. PMID- 12588711 TI - Broadening the therapeutic options in acute asthma. PMID- 12588712 TI - Ischemia-reperfusion-induced lung injury. AB - Ischemia-reperfusion-induced lung injury is characterized by nonspecific alveolar damage, lung edema, and hypoxemia occurring within 72 hours after lung transplantation. The most severe form may lead to primary graft failure and remains a significant cause of morbidity and mortality after lung transplantation. Over the past decade, better understanding of the mechanisms of ischemia-reperfusion injury, improvements in the technique of lung preservation, and the development of a new preservation solution specifically for the lung have been associated with a reduction in the incidence of primary graft failure from approximately 30 to 15% or less. Several strategies have also been introduced into clinical practice for the prevention and treatment of ischemia-reperfusion induced lung injury with various degrees of success. However, only three randomized, double-blinded, placebo-controlled trials on ischemia-reperfusion induced lung injury have been reported in the literature. In the future, the development of new agents and their application in prospective clinical trials are to be expected to prevent the occurrence of this potentially devastating complication and to further improve the success of lung transplantation. PMID- 12588713 TI - Pulmonary matrix metalloproteinase excess in hospital-acquired pneumonia. AB - In hospital-acquired pneumonia, extracellular matrix destruction is common and may be caused by excessive activity of matrix metalloproteinases (MMPs). Thirty patients with hospital-acquired pneumonia and 16 control subjects were studied. We evaluated the concentrations of MMP-8, MMP-9, and tissue inhibitor of metalloproteinase-1 in mini-bronchoalveolar lavage fluid (mini-BALF) and blood using zymography and specific immunoassays. In patients with hospital-acquired pneumonia concentrations of MMP-8 and MMP-9 in mini-BALF were increased 10-fold, whereas their specific inhibitor tissue inhibitor of metalloproteinase-1 was not concomitantly increased. In 80% of patients with pneumonia, but in none of the control subjects, the active form of MMP-9 was detected by zymography. Zymography furthermore showed the banding pattern of neutrophil-derived MMP-9, indicating that neutrophils were the main source of MMP-9. Comparison of neutrophils from blood and mini-BALF showed higher basal release of MMPs by pulmonary neutrophils. Stimulation analysis indicated that pulmonary neutrophils were already maximally activated. In patients with detection of potentially pathogenic microorganisms, concentrations of MMPs were fivefold increased compared with patients with negative cultures. Furthermore, MMP-levels were related to clinical severity. These are the first data suggesting that neutrophil-derived MMPs are increased in hospital-acquired pneumonia in association to the detection of causative microorganisms and clinical severity. PMID- 12588714 TI - American Thoracic Society/Centers for Disease Control and Prevention/Infectious Diseases Society of America: treatment of tuberculosis. PMID- 12588715 TI - Bactericidal activity of antituberculosis drugs. PMID- 12588716 TI - A reiterative method for calculating bactericidal activity. PMID- 12588717 TI - Cardiopulmonary resuscitation in pulmonary hypertension. PMID- 12588718 TI - Exhaled nitric oxide as a diagnostic tool. PMID- 12588719 TI - Time for standardization. PMID- 12588720 TI - Growth habit and mechanical architecture of the sand dune-adapted climber Clematis flammula var. maritima L. AB - Clematis flammula var. maritima is a woody lianoid plant that grows on coastal sand dunes in the Mediterranean region. Older perennial stems are present as extensive underground axes. These generate surface growth of shorter-lived stems producing monospecific trellises above the surface of the sand. Despite its sand dune habitat and shortage of host support plants, this variety of Clematis shows mechanical characteristics during growth that are closely comparable with those of scandent woody lianas. A significant decrease in the value of structural Young's modulus is observed from the aerial trellis-forming shoots (1.619 +/- 0.492 GN m(-2)) to emergent axes (0.855 +/- 0.253 GN m(-2)) and underground woody stems (0.470 +/- 0.113 GN m(-2)). Biomechanical and developmental observations indicate that most emergent branches are optimized geometrically and mechanically in relation to their points of emergence from the sand, with increases in structural Young's modulus and the second moment of area around the surface of the sand. Lianoid plants, physiologically capable of withstanding sand dune environments, might represent acceptable natural or introduced species for dune stabilization and conservation. PMID- 12588721 TI - Refilling of a hydraulically isolated embolized xylem vessel: model calculations. AB - When they are hydraulically isolated, embolized xylem vessels can be refilled, while adjacent vessels remain under tension. This implies that the pressure of water in the refilling vessel must be equal to the bubble gas pressure, which sets physical constraints for recovery. A model of water exudation into the cylindrical vessel and of bubble dissolution based on the assumption of hydraulic isolation is developed. Refilling is made possible by the turgor of the living cells adjacent to the refilling vessel, and by a reflection coefficient below 1 for the exchange of solutes across the interface between the vessel and the adjacent cells. No active transport of solutes is assumed. Living cells are also capable of importing water from the water-conducting vessels. The most limiting factors were found to be the osmotic potential of living cells and the ratio of the volume of the adjacent living cells to that of the embolized vessel. With values for these of 1.5 MPa and 1, respectively, refilling times were in the order of hours for a broad range of possible values of water conductivity coefficients and effective diffusion distances for dissolved air, when the xylem water tension was below 0.6 MPa and constant. Inclusion of the daily pattern for xylem tension improved the simulations. The simulated gas pressure within the refilling vessel was in accordance with recent experimental results. The study shows that the refilling process is physically possible under hydraulic isolation, while water in surrounding vessels is under negative pressure. However, the osmotic potentials in the refilling vessel tend to be large (in the order of 1 MPa). Only if the xylem water tension is, at most, twice atmospheric pressure, the reflection coefficient remains close to 1 (0.95) and the ratio of the volume of the adjacent living cells to that of the embolized vessel is about 2, does the osmotic potential stay below 0.4 MPa. PMID- 12588722 TI - Time-course of tomato whole-plant respiration and fruit and stem growth during prolonged darkness in relation to carbohydrate reserves. AB - To evaluate the relevance of a simple carbon balance model (Seginer et al., 1994, Scientia Horticulturae 60: 55-80) in source-limiting conditions, the dynamics of growth, respiration and carbohydrate reserves of tomato plants were observed in prolonged darkness. Four days prior to the experiments, plants were exposed to high or low light levels and CO(2) concentrations. The concentration of carbohydrates in vegetative organs was 30-50 % lower in plants that were exposed to low carbon assimilation conditions compared with those exposed to high carbon assimilation conditions. During prolonged darkness, plants with low carbohydrate reserves exhibited a lower whole-plant respiration rate, which decreased rapidly to almost zero after 24 h, and carbohydrate pools were almost exhausted in leaves, roots and flowers. In plants with high carbohydrate reserves, the whole plant respiration rate was maintained for a longer period and carbohydrates remained available for at least 48 h in leaves and flowers. In contrast, fruits maintained fairly stable and identical concentrations of carbohydrates and the reduction in their rate of expansion was moderate irrespective of the pre treatment carbon assimilation conditions. The time-course of asparagine and glutamine concentrations showed the occurrence of carbon stress in leaves and flowers. Estimation of source and sink activities indicated that even after low carbon assimilation, vegetative organs contained enough carbohydrates to support fruit growth provided their own growth stopped. The time of exhaustion of these carbohydrates corresponded grossly to the maintenance stage simulated by the model proposed by Seginer et al. (1994), thus validating the use of such a model for optimizing plant growth. PMID- 12588723 TI - Lipoidal labellar secretions in Maxillaria ruiz & pav. (Orchidaceae). AB - The labella of Maxillaria acuminata Lindl., M. cerifera Barb. Rodr. and M. notylioglossa Rchb.f., all members of the M. acuminata alliance, produce a viscid wax-like secretion. Histochemical analysis revealed that the chemical composition of the secretion is similar in all three species, consisting largely of lipid and protein. Light microscopy and low-vacuum scanning electron microscopy were used to investigate the secretory process. In a fourth taxon, M. cf. notylioglossa, transmission electron microscopy showed that lipid bodies are associated with smooth endoplasmic reticulum or occur as plastoglobuli within plastids. Lipid bodies vary in appearance and this may reflect differences in chemical composition. They become associated with the plasmalemma and eventually accumulate between the latter and the cell wall. The wall contains no pits or ectodesmata, and it is speculated that lipid passes through the wall as small lipid moieties before eventually reassembling to form lipid globules on the external surface of the cuticle. These globules are able to coalesce forming extensive viscid areas on the labellum. The possible significance of this process to pollination is discussed. PMID- 12588724 TI - Responses of soybean to oxygen deficiency and elevated root-zone carbon dioxide concentration. AB - Root flooding is damaging to the growth of crop plants such as soybean (Glycine max L.). Field flooding for 3 d often results in leaf chlorosis, defoliation, cessation of growth and plant death. These effects have been widely attributed solely to a lack of oxygen in the root-zone. However, an additional damaging factor may be CO(2), which attains levels of 30 % (v/v) of total dissolved gases. Accordingly, the effects of root-zone CO(2) on oxygen-deficient soybean plants were investigated in hydroponic culture. Soybean plants are shown to be very tolerant of excess water and anaerobiosis. No oxygen (100 % N(2) gas) and low oxygen (non-aerated) treatments for 14 d had no effect on soybean survival or leaf greenness, but plants became severely chlorotic and stunted when the roots were exposed to no oxygen together with CO(2) concentrations similar to those in flooded fields (equilibrium concentrations of 30 %). When root-zone CO(2) was increased to 50 %, a quarter of soybean plants died. Those plants that survived showed severe symptoms of chlorosis, necrosis and root death. In contrast, rice (Oryza sativa L.) plants were not affected by the combination of no oxygen and elevated root-zone CO(2.) A concentration of 50 % CO(2) did not affect rice plant survival or leaf colour. These results suggest that the high susceptibility of soybean to soil flooding, compared with that of rice, is an outcome of its greater sensitivity to CO(2). PMID- 12588725 TI - Effects of plant traits on ecosystem and regional processes: a conceptual framework for predicting the consequences of global change. AB - Human activities are causing widespread changes in the species composition of natural and managed ecosystems, but the consequences of these changes are poorly understood. This paper presents a conceptual framework for predicting the ecosystem and regional consequences of changes in plant species composition. Changes in species composition have greatest ecological effects when they modify the ecological factors that directly control (and respond to) ecosystem processes. These interactive controls include: functional types of organisms present in the ecosystem; soil resources used by organisms to grow and reproduce; modulators such as microclimate that influence the activity of organisms; disturbance regime; and human activities. Plant traits related to size and growth rate are particularly important because they determine the productive capacity of vegetation and the rates of decomposition and nitrogen mineralization. Because the same plant traits affect most key processes in the cycling of carbon and nutrients, changes in plant traits tend to affect most biogeochemical cycling processes in parallel. Plant traits also have landscape and regional effects through their effects on water and energy exchange and disturbance regime. PMID- 12588726 TI - Inhibition of the indole-3-acetic acid-induced epinastic curvature in tobacco leaf strips by 2,4-dichlorophenoxyacetic acid. AB - It has been reported that auxin induces an epinastic growth response in plant leaf tissues. Leaf strips of tobacco (Nicotiana tabacum L. 'Bright Yellow 2') were used to study the effects of indole-3-acetic acid (IAA), the principal form of auxin in higher plants, and a synthetic auxin, 2,4-dichlorophenoxyacetic acid (2,4-D), on epinastic leaf curvature. Incubation of leaf strips with 10 micro M IAA resulted in a marked epinastic curvature response. Unexpectedly, 2,4-D showed only a weak IAA-like activity in inducing epinasty. Interestingly, the presence of 2,4-D resulted in inhibition of the IAA-dependent epinastic curvature. In vivo Lineweaver-Burk kinetic analysis clearly indicated that the interaction between IAA and 2,4-D reported here is not a result of competitive inhibition. Using kinetic analysis, it was not possible to determine whether the mode of interaction between IAA and 2,4-D was non-competitive or uncompetitive. 2,4-D inhibits the IAA-dependent epinasty via complex and as yet unidentified mechanisms. PMID- 12588727 TI - Characteristics of floral organs related to reliable self-pollination in rice (Oryza sativa L.). AB - In many varieties of rice, the length of basal pores on the thecae just after anthesis is strongly correlated both with the percentage of florets receiving adequate pollen and with the number of pollen grains deposited on the stigmata and its variation (coefficient of variation). Therefore, the size of the basal pores is considered to be an important factor for the reliable self-pollination of rice. We discuss how long basal pores may facilitate self- pollination. PMID- 12588728 TI - Analysing branching pattern in plantations of young red oak trees (Quercus rubra L., Fagaceae). AB - Branching patterns of the growth units of monocyclic or bicyclic annual shoots on the main axis of 5-year-old red oaks were studied in a plantation in south western France. For each growth unit, the production of axillary structures associated with each node was described in the form of a sequence. For a given category of growth units, homogeneous zones (i.e. zones in which composition in terms of type of axillary production does not change substantially) were identified on such sequences using a dedicated statistical model called a hidden semi-Markov chain. For instance, on the first growth unit of bicyclic annual shoots, a zone with 1-year-delayed branches was found systematically below a zone with buds and one-cycle-delayed branches. Branching patterns shown by the growth unit of monocyclic annual shoots and on the second growth unit of bicyclic annual shoots were very similar. Branches with a 1-year delay in development tended to be polycyclic at the top of the growth unit and monocyclic lower down. The number of nodes shown by the branched zone of the growth unit of monocyclic annual shoots was stable, irrespective of the total number of nodes of the growth unit. In contrast, the second growth unit of bicyclic annual shoots exhibited a correlation between the number of nodes in the branching zone and the total number of nodes. The contribution made by this method to understanding plant functioning is discussed. PMID- 12588729 TI - Variability of the essential oil of Viola etrusca. AB - Essential oils obtained from different populations of Viola etrusca from Italy have been analysed to verify the phenotypic discontinuity observed in a previous study. All of the essential oils contained methyl salicylate as a main constituent. However, multivariate analysis showed differences among some populations, in particular between northern and southern ones. Results suggest that this species could be undergoing a slow schizogenetic differentiation process due to its genetic isolation. PMID- 12588732 TI - Sensitivity-dependent hierarchical receptor codes for odors. AB - In order to comprehend the strategy of odor encoding by odorant receptors, we isolated 2740 mouse receptor neurons from four olfactory epithelial zones and classified them in terms of their sensitivities and tuning specificities to a chiral pair of odorants, S(+)-carvone (caraway-like odor) and R(-)-carvone (spearmint-like odor). Our approach revealed that the majority of receptors at the lowest effective stimulus concentration represented the principal odor qualities characteristic of each enantiomer by means of the principal odor qualities of the odorants for which the receptors were most sensitive. The chiral non-discriminating receptors were newly recruited 3.7 times of R(-)-carvone sensitive receptors and totally became 2.8 times (39/14) of R(-)carvone-sensitive receptors in the subpopulations when the stimulus concentration was increased 10 fold [corrected]. More than 80% of the responsive receptors (an estimated 70 +/- alpha types) exhibited overlapping sensitivities between the enantiomers. The signals from the non-discriminating receptors may be reduced to decode the characteristic odor identity for R(-)-carvone in the brain over an adequate range of stimulus strengths. The information processing of odors appears to involve the selective weighting of the signals from the most sensitive receptors. An analysis of the overall receptor codes to carvones indicated that the system employs hierarchical receptor codes: principal odor qualities are encoded by the most sensitive receptors and lower-ranked odor qualities by less sensitive receptors. PMID- 12588733 TI - SURE, why not? The SUbstitution-REciprocity method for measurement of odor quality discrimination thresholds: replication and extension to nonhuman primates. AB - Recently, Olsson and Cain (2000, Chem. Senses, 25: 493) introduced a psychometric method which, for the first time, allows the standardized determination of odor quality discrimination (OQD) thresholds. The method defines a threshold value that is an average fraction by which one odorant has to be substituted with another to reach a criterion level of discrimination. This measure of discrimination is reciprocal in the sense that it is a result of two separate psychometric functions involving two different standards but the same comparison stimuli. Using the same odor stimuli as Olsson and Cain, with six human subjects but adopting a slightly different experimental design, we were able to replicate their finding that the proportion of correct discriminations changes monotonically with the proportion of adulterant in mixtures of eugenol and citral. As the SURE (SUbstitution-REciprocity) method is based on discriminative responses, it should also be applicable with nonhuman species which can be trained to give unequivocal discriminative responses at the behavioral level. Using an olfactory conditioning paradigm, we therefore trained four squirrel monkeys to discriminate between exactly the same pairs of odor stimuli as our human subjects. We found the psychometric functions of the monkeys to be similar to those of the human subjects. Our results show that the SURE method can successfully be employed with nonhuman primates and thus offers a new approach to study the odor spaces of nonhuman species. Future studies should elucidate whether the SURE method allows for direct comparisons of OQD thresholds and of similarities and differences between odor quality perception of different species. PMID- 12588734 TI - Projections to higher olfactory centers from subdivisions of the antennal lobe macroglomerular complex of the male silkmoth. AB - The macroglomerular complex (MGC) is the first-order center for synaptic processing of olfactory information about the female sex pheromone in the male moth brain. We have investigated the MGC subdivisions of the male silkmoth Bombyx mori by use of three-dimensional reconstruction of the MGC from sequential series of confocal slice images. The B. mori MGC consists of three subdivisions similar to those of Manduca sexta: the cumulus, toroid and horseshoe. Intracellular recording and staining revealed that responses of MGC projection neurons to pheromonal stimulation correlate with their dendritic arborizations in the subdivisions of the MGC (the cumulus, toroid and horseshoe) and each subdivision specific projection neuron transmits information to different regions in the calyces of the mushroom body and the inferior lateral protocerebrum. We revealed that major pheromone component information is transferred to the medial part of the inferior lateral protocerebrum through three different antennocerebral pathways. Although it is generally accepted that the calyces of the mushroom body and the inferior lateral protocerebrum are the target sites for pheromone information from the MGC in moths, our results suggest that the medial part of the inferior lateral protocerebrum may be a more important processing site for major pheromonal information in B. mori. PMID- 12588736 TI - Inducible defences in Daphnia depend on latent alarm signals from conspecific prey activated in predators. AB - Some water fleas (Daphnia spp.) undergo phenotypic changes when exposed to chemical signals from predators. The chemical signals have been assumed to be of predator origin (i.e. kairomones), since juices of crushed Daphnia have been found ineffective. We speculated that latent alarm signals could be present in Daphnia, to be activated in predators following ingestion. Accordingly, fish predators were fed earthworms for 10 weeks to remove Daphnia remains from their gastro-intestinal tracts. Following another 6 days of earthworm feeding, water conditioned by fish induced no morphological changes in D. galeata. When fish were alternatively fed Daphnia for 6 days, changes were induced with fish conditioned water. Extracts made from intestines of earthworm-fed fish, homogenized with earthworms, gave no morphological changes, but intestines of the same origin homogenized with Daphnia did. Similar results were found when earthworms and Daphnia were homogenized with fish liver. Freshly frozen extracts of homogenized Daphnia gave no detectable changes at first instar stage in test animals, whereas extracts of Daphnia that had been kept at room temperature did induce such changes. Our results suggest that Daphnia respond to latent conspecific alarm signals (i.e. 'dormant' pheromones) that are activated by intestinal or bacterial enzymes in predators or in the water. PMID- 12588735 TI - Responsiveness of the cortical taste area neurons to a mixture of the four basic tastants in rats. AB - The taste coding mechanism in the cortical taste area was investigated by analyzing the responses of 59 neurons in the cortical taste area of the anesthetized rat to a mixture of the four basic tastants in both absence and presence of bicuculline methiodide, a specific antagonist to the GABA(A) receptors. The mixture caused response suppression more frequently than response facilitation, both in the control state and during bicuculline application. Cluster analysis revealed that only a group of the neurons with the best response to both NaCl and HCl (group NH) showed the best response to the mixture in the control state, whereas during bicuculline application, in addition to group NH, two other groups of neurons responding to sucrose, or to HCl and quinine responded vigorously to the mixture. Multidimensional scaling located the mixture outside the space of the four basic tastants facing an NaCl-HCl line in both states. GABAergic inhibition caused the group NH to represent the taste of the mixture in the control state. Thus, the mixture probably tastes salty and sour to rats. No cortical neuron was found which specifically responded to the mixture. PMID- 12588737 TI - Descending influences from the lateral hypothalamus and amygdala converge onto medullary taste neurons. AB - The lateral hypothalamus (LH) and the central nucleus of the amygdala (CeA) exert an influence on many aspects of ingestive behavior. These nuclei receive projections from several areas carrying gustatory and viscerosensory information, and send axons to these nuclei as well, including the nucleus of the solitary tract (NST). Gustatory responses of NST neurons are modulated by stimulation of the LH and the CeA, and by several physiological factors related to ingestive behavior. We investigated the effect of both LH and CeA stimulation on the activity of 215 taste-responsive neurons in the hamster NST. More than half of these neurons (113/215) were modulated by electrical stimulation of the LH and/or CeA; of these, 52 cells were influenced by both areas, often bilaterally. The LH influenced more neurons than the CeA (101 versus 64 cells). Contralateral stimulation of these forebrain areas was more often effective (144 responses) than ipsilateral (74). Modulatory effects were mostly excitatory (102 cells); 11 cells were inhibited, mostly by ipsilateral LH stimulation. A subset of these cells (n = 25) was examined for the effects of microinjection of DL-homocysteic acid (DLH), a glutamate receptor agonist, into the LH and/or CeA. The effects of electrical stimulation were completely mimicked by DLH, indicating that cell somata in and around the stimulating sites were responsible for these effects. Other cells (n = 25) were tested for the effects of electrical stimulation of the LH and/or CeA on the responses to taste stimulation of the tongue (32 mM sucrose, NaCl and quinine hydrochloride, and 3.2 mM citric acid). Responses to taste stimuli were enhanced by the excitatory influence of the LH and/or CeA. These data demonstrate that descending influences from the LH and CeA reach many of the same cells in the gustatory NST and can modulate their responses to taste stimulation. PMID- 12588738 TI - Heightened sour preferences during childhood. AB - Basic research has revealed that the chemical sensory world of children is different from that of adults, as evidenced by their heightened preferences for sweet and salty tastes. However, little is known about the ontogeny of sour taste preferences, despite the growing market of extreme sour candies. The present study investigated whether the level of sourness most preferred in a food matrix and the ability to discriminate differences in sour intensity differed between 5- to 9-year-old children and their mothers, by using a rank-by-elimination procedure embedded in the context of a game. Mothers also completed a variety of questionnaires and children were asked several questions to assess whether children's temperament and food preferences and habits related to sour preferences. The results indicated that, although every mother and all but two of the children (92%) were able to rank the gelatins from most to least sour, more than one-third (35%) of the children, but virtually none of the adults, preferred the high levels of sour taste (0.25 M citric acid) in gelatin. Those children who preferred the extreme sour tastes were significantly less food neophobic (P < 0.05) and tended to experience a greater variety of fruits when compared with the remaining children (P = 0.11). Moreover, the children's preference for sour tastes generalized to other foods, such as candies and lemons, as reported by both children and mothers. These findings are the first experimental evidence to demonstrate that sour taste preferences are heightened during childhood and that such preferences are related to children's food habits and preferences. Further research is needed to unfold the relationship between the level of sour taste preferred and the actual consumption of sour-tasting foods and flavors in children. PMID- 12588739 TI - Iron-dependent hydrogenases of Entamoeba histolytica and Giardia lamblia: activity of the recombinant entamoebic enzyme and evidence for lateral gene transfer. AB - Entamoeba histolytica and Spironucleus barkhanus have genes that encode short iron-dependent hydrogenases (Fe-hydrogenases), even though these protists lack hydrogenosomes. To understand better the biochemistry of the protist Fe hydrogenases, we prepared a recombinant E. histolytica short Fe-hydrogenase and measured its activity in vitro. A Giardia lamblia gene encoding a short Fe hydrogenase was identified from shotgun genomic sequences, and RT-PCR showed that cultured entamoebas and giardias transcribe short Fe-hydrogenase mRNAs. A second E. histolytica gene, which encoded a long Fe-hydrogenase, was identified from shotgun genomic sequences. Phylogenetic analyses suggested that the short Fe hydrogenase genes of entamoeba and diplomonads share a common ancestor, while the long Fe-hydrogenase gene of entamoeba appears to have been laterally transferred from a bacterium. These results are discussed in the context of competing ideas for the origins of genes encoding fermentation enzymes of these protists. PMID- 12588740 TI - Molecular genetic evidence that dinoflagellates belonging to the genus Symbiodinium freudenthal are haploid. AB - Microscopic and cytological evidence suggest that many dinoflagellates possess a haploid nuclear phase. However, the ploidy of a number of dinoflagellates remains unknown, and molecular genetic support for haploidy in this group has been lacking. To elucidate the ploidy of symbiotic dinoflagellates belonging to the genus Symbiodinium, we used five polymorphic microsatellites to examine populations harbored by the Caribbean gorgonians Plexaura kuna and Pseudopterogorgia elisabethae; we also studied a series of Symbiodinium cultures. In 690 out of 728 Symbiodinium samples in hospite (95% of the cases) and in all 45 Symbiodinium cultures, only a single allele was recovered per locus. Statistical testing of the Symbiodinium populations harbored by P. elisabethae revealed that the observed genotype frequencies deviate significantly from those expected under Hardy-Weinberg equilibrium. Taken together, our results confirm that, in the vegetative life stage, members of Symbiodinium, both cultured and in hospite, are haploid. Furthermore, based on the phylogenetics of the dinoflagellates, haploidy in vegetative cells appears to be an ancestral trait that extends to all 2,000 extant species of these important unicellular protists. PMID- 12588741 TI - Amebocyte production begins at stage 18 during embryogenesis in Limulus polyphemus, the American horseshoe crab. AB - Limulus polyphemus, the American horseshoe crab, has a single type of circulating blood cell, the granular amebocyte, which is the horseshoe crab's primary cellular defense against microbial infection. On exposure to gram-negative bacteria or their endotoxins, the amebocytes degranulate, releasing the clotting protein coagulogen and a number of proteases. The protease cascade converts the soluble coagulogen to insoluble coagulin, which forms fibrous clots that seal off the site of infection. The first description of this clotting reaction in the 1950s initiated development of Limulus amebocyte lysate and spurred an intensive study of the amebocytes. However, the site or sites and timing of amebocyte production have yet to be determined. We report here that during embryonic development in Limulus polyphemus, amebocyte production begins at stage 18. The first amebocytes detected are found in developing hemocoel cavities, and the cells may derive from previously undifferentiated yolk nuclei. PMID- 12588742 TI - High-speed video analysis of the escape responses of the copepod Acartia tonsa to shadows. AB - The copepod Acartia tonsa exhibits a vigorous escape jump in response to rapid decreases in light intensity, such as those produced by the shadow of an object passing above it. In the laboratory, decreases in light intensity were produced using a fiber optic lamp and an electronic shutter to abruptly either nearly eliminate visible light or reduce light intensity to a constant proportion of its original intensity. The escape responses of A. tonsa to these rapid decreases in visible light were recorded on high-speed video using infrared illumination. The speed, acceleration, and direction of movement of the escape response were quantified from videotape by using automated motion analysis techniques. A. tonsa typically responds to decreases in light intensity with an escape jump comprising an initial reorientation followed by multiple power strokes of the swimming legs. These escape jumps can result in maximum speeds of over 800 mm s(-1) and maximum accelerations of over 200 m s(-2). In A. tonsa, photically stimulated escape responses differ from hydrodynamically stimulated responses mainly in the longer latencies of photically stimulated responses and in the increased number of power strokes, even when the stimulus is near threshold; these factors result in longer escape jumps covering greater distances. The latency of responses of A. tonsa to this photic stimulus ranged from a minimum of about 30 ms to a maximum of more than 150 ms, compared to about 4 ms for hydrodynamically stimulated escape jumps. Average response latency decreased with increasing light intensity or increasing proportion of light eliminated. Little change was observed in the vigor of the escape response to rapid decreases in visible light over a wide range of adaptation intensities. PMID- 12588743 TI - Behavioral thermoregulation in Hemigrapsus nudus, the amphibious purple shore crab. AB - The thermoregulatory behavior of Hemigrapsus nudus, the amphibious purple shore crab, was examined in both aquatic and aerial environments. Crabs warmed and cooled more rapidly in water than in air. Acclimation in water of 16 degrees C (summer temperatures) raised the critical thermal maximum temperature (CTMax); acclimation in water of 10 degrees C (winter temperatures) lowered the critical thermal minimum temperature (CTMin). The changes occurred in both water and air. However, these survival regimes did not reflect the thermal preferences of the animals. In water, the thermal preference of crabs acclimated to 16 degrees C was 14.6 degrees C, and they avoided water warmer than 25.5 degrees C. These values were significantly lower than those of the crabs acclimated to 10 degrees C; these animals demonstrated temperature preferences for water that was 17 degrees C, and they avoided water that was warmer than 26.9 degrees C. This temperature preference was also exhibited in air, where 10 degrees C acclimated crabs exited from under rocks at a temperature that was 3.2 degrees C higher than that at which the 16 degrees C acclimated animals responded. This behavioral pattern was possibly due to a decreased thermal tolerance of 16 degrees C acclimated crabs, related with the molting process. H. nudus was better able to survive prolonged exposure to cold temperatures than to warm temperatures, and there was a trend towards lower exit temperatures with the lower acclimation (10 degrees C) temperature. Using a complex series of behaviors, the crabs were able to precisely control body temperature independent of the medium, by shuttling between air and water. The time spent in either air or water was influenced more strongly by the temperature than by the medium. In the field, this species may experience ranges in temperatures of up to 20 degrees C; however, it is able to utilize thermal microhabitats underneath rocks to maintain its body temperature within fairly narrow limits. PMID- 12588744 TI - Synthesis of a high-density lipoprotein in the developing blue crab (Callinectes sapidus). AB - An important lipoprotein in the hemolymph of crustaceans is LpI. It transports lipid to peripheral tissues and also has a role in crustacean immune recognition. We employed a monoclonal antibody specific for the LpI peptide to demonstrate by ELISA, western blot and immunohistochemistry the appearance of LpI during development of Callinectes sapidus, the blue crab. LpI was first found in stage 5 embryos and appeared to be synthesized by lateral basophilic cuboidal cells that demonstrated cytoplasmic immunoreactivity for LpI at their interface with the yolk mass. The embryonic cuboidal cells bore a strong cytologic resemblance to the hepatopancreas cells of later stages (zoea, megalopae, adults), which were also immunoreactive for LpI. PMID- 12588745 TI - Reproductive biology of Hemiramphus brasiliensis and H. balao (hemiramphidae): maturation, spawning frequency, and fecundity. AB - Analyses of life-history data show that both the size-specific batch fecundities and the age-specific spawning frequencies differ for two halfbeak species, Hemiramphus brasiliensis, the ballyhoo, and H. balao, the balao. Halfbeak ages were determined from sectioned otoliths; histological data was used to describe oocyte development and estimate spawning frequency; and batch fecundity was measured from counts of whole oocytes in final maturation. Hemiramphus brasiliensis lived longer (4 versus 2 years) and had a higher survival rate (14.9% versus 7.5% annually) than H. balao did. Of the two species the larger and longer-lived congener, H. brasiliensis, reached sexual maturity at a larger size (fork length 198 versus 160 mm). The spawning period of age-0 females was strongly related to season, whereas spawning by older females occurred throughout the year. Reproduction by both species peaked during late spring or early summer, and all mature females were spawning daily during April (H. brasiliensis) or June (H. balao). This is the first demonstration of iteroparity for the family Hemiramphidae. H. brasiliensis had a lower batch fecundity (about 1164 versus 3743 hydrated oocytes for a 100-g female) than H. balao did. Such low batch fecundities are typical of the order Beloniformes, but quite different from those of other fishes that live in association with coral reef habitats. H. balao's higher batch fecundity is consistent with the life-history theory that predicts higher numbers of eggs for shorter-lived species; this is possible because H. balao produces smaller hydrated oocytes than H. brasiliensis (modal diameter about 1.6 versus 2.4 mm). The high spawning frequency of Hemiramphus species compensates for their low batch fecundity. The annual fecundity of both species is similar to that of other reef fish species, after adjusting for body size and spawning frequency. The lifetime fecundity of H. balao was very similar to that of H. brasiliensis, after accounting for the differences in survival for each species. This suggests a fine tuning of different reproductive traits over the entire life cycle that results in roughly equivalent lifetime fecundity for both species. PMID- 12588746 TI - Collection and culture techniques for gelatinous zooplankton. AB - Gelatinous zooplankton are the least understood of all planktonic animal groups. This is partly due to their fragility, which typically precludes the capture of intact specimens with nets or trawls. Specialized tools and techniques have been developed that allow researchers and aquarists to collect intact gelatinous animals at sea and to maintain many of these alive in the laboratory. This paper summarizes the scientific literature on the capture, collection, and culture of gelatinous zooplankton and incorporates many unpublished methods developed at the Monterey Bay Aquarium in the past 15 years. PMID- 12588747 TI - Branchial musculature of a venerid clam: pharmacology, distribution, and innervation. AB - This study was meant to analyze the neural control of the branchial muscles of the clam Mercenaria mercenaria. Gills isolated from the animal contract in response to 5-hydroxytryptamine (5HT), dopamine (DA), and acetylcholine (ACh); but the ACh contraction occurred only if the gills had been pretreated with the cholinesterase inhibitor eserine. The 5HT antagonists cyproheptadine and mianserin blocked the contractile effects of all of the agonists. However, gills exposed to the 5HT antagonists and eserine relaxed in response to ACh. The DA antagonist SCH-83566 inhibited the effects of DA, but had no effect on contractions induced by 5HT and ACh. The ACh antagonist hexamethonium inhibited both the excitatory and inhibitory effects of ACh, but had no effect on contractions induced by 5HT and DA. 5HT and DA in gill tissue were visualized by using immunohistochemistry. Within each gill filament are dorsoventral neurons running adjacent to the epithelium and containing immunoreactive 5HT and DA. A complex network of 5HT-positive fibers is associated with the septa, blood vessels, and muscles, whereas DA-positive fibers are restricted to the septa. We propose that 5HT is the excitatory transmitter to the gill muscles, and that DA and ACh exert their excitatory effects by stimulating 5HT motor nerves. ACh may also be an inhibitory transmitter of the muscles. PMID- 12588748 TI - Salinity tolerance of larval Rapana venosa: implications for dispersal and establishment of an invading predatory gastropod on the North American Atlantic coast. AB - The lack of quantitative data on the environmental tolerances of the early life history stages of invading species hinders estimation of their dispersal rates and establishment ranges in receptor environments. We present data on salinity tolerance for all stages of the ontogenetic larval development of the invading predatory gastropod Rapana venosa, and we propose that salinity tolerance is the dominant response controlling the potential dispersal (=invasion) range of the species into the estuaries of the Atlantic coast of the United States from the current invading epicenter in the southern Chesapeake Bay. All larval stages exhibit 48-h tolerance to salinities as low as 15 ppt with minimal mortality. Below this salinity, survival grades to lower values. Percentage survival of R. venosa veligers was significantly less at 7 ppt than at any other salinity. There were no differences in percentage survival at salinities greater than 16 ppt. We predict that the counterclockwise, gyre-like circulation within the Chesapeake Bay will initially distribute larvae northward along the western side of the DelMarVa peninsula, and eventually to the lower sections of all major subestuaries of the western shore of the Bay. Given the observed salinity tolerances and the potential for dispersal of planktonic larvae by coastal currents, establishment of this animal over a period of decades from Cape Cod to Cape Hatteras is a high probability. PMID- 12588749 TI - Short-distance spawning migration of tropical freshwater eels. PMID- 12588750 TI - Omega-3 fatty acids and cardiovascular disease: new recommendations from the American Heart Association. PMID- 12588751 TI - Triglyceride-rich lipoproteins and vascular function. PMID- 12588752 TI - Rapid effects of statins: from prophylaxis to therapy for ischemic stroke. PMID- 12588753 TI - Advantages of fast-acting ADP receptor blockade in ischemic heart disease. PMID- 12588754 TI - Cholesteryl ester transfer protein: a novel target for raising HDL and inhibiting atherosclerosis. AB - Cholesteryl ester transfer protein (CETP) promotes the transfer of cholesteryl esters from antiatherogenic HDLs to proatherogenic apolipoprotein B (apoB) containing lipoproteins, including VLDLs, VLDL remnants, IDLs, and LDLs. A deficiency of CETP is associated with increased HDL levels and decreased LDL levels, a profile that is typically antiatherogenic. Studies in rabbits, a species with naturally high levels of CETP, support the therapeutic potential of CETP inhibition as an approach to retarding atherogenesis. Studies in mice, a species that lacks CETP activity, have provided mixed results. Human subjects with heterozygous CETP deficiency and an HDL cholesterol level >60 mg/dL have a reduced risk of coronary heart disease. Evidence that atherosclerosis may be increased in CETP-deficient subjects whose HDL levels are not increased is difficult to interpret and may reflect confounding or bias. Small-molecule inhibitors of CETP have now been tested in human subjects and shown to increase the concentration of HDL cholesterol while decreasing that of LDL cholesterol and apoB. Thus, it seems important and timely to test the hypothesis in randomized trials of humans that pharmacological inhibition of CETP retards the development of atherosclerosis. PMID- 12588755 TI - Endothelial dysfunction: a marker of atherosclerotic risk. AB - Endothelial dysfunction is a systemic disorder and a key variable in the pathogenesis of atherosclerosis and its complications. Current evidence suggests that endothelial status is not determined solely by the individual risk factor burden but rather, may be regarded as an integrated index of all atherogenic and atheroprotective factors present in an individual, including known as well as yet unknown variables and genetic predisposition. Endothelial dysfunction reflects a vascular phenotype prone to atherogenesis and may therefore serve as a marker of the inherent atherosclerotic risk in an individual. In line with this hypothesis, dysfunction of either the coronary or peripheral vascular endothelium was shown to constitute an independent predictor of cardiovascular events, providing valuable prognostic information additional to that derived from conventional risk factor assessment. Interventions like risk factor modification and treatment with various drugs, including statins and angiotensin-converting enzyme inhibitors, may improve endothelial function and thereby, potentially prognosis. Hence, given its reversibility and granted the availability of a diagnostic tool to identify patients at risk and to control the efficacy of therapy in clinical practice, endothelial dysfunction may be an attractive primary target in the effort to optimize individualized therapeutic strategies to reduce cardiovascular morbidity and mortality. PMID- 12588756 TI - HMG-CoA reductase inhibitor increases GTP cyclohydrolase I mRNA and tetrahydrobiopterin in vascular endothelial cells. AB - OBJECTIVE: Endothelial nitric oxide synthase (eNOS) activity is supported by tetrahydrobiopterin (BH4), which appears to be important for generating protective NO but decreases uncoupling formation of superoxide. We investigated the effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, or statins, in terms of BH4 metabolism in human umbilical vein endothelial cells (HUVECs). METHODS AND RESULTS: We measured the mRNA levels of GTP cyclohydrolase I (GTPCH), the rate-limiting enzyme in the first step of de novo BH4 synthesis, by real-time polymerase chain reaction. The mRNA of GTPCH, as well as of eNOS, was upregulated in HUVECs treated with cerivastatin. This increase was time and dose dependent. Fluvastatin was also observed to enhance GTPCH and eNOS mRNA levels. In parallel with this observation, cerivastatin increased intracellular BH4. Incubating HUVECs with tumor necrosis factor (TNF-alpha) was observed to increase GTPCH mRNA while decreasing eNOS mRNA. In the presence of cerivastatin, the TNF-alpha-mediated increase in GTPCH mRNA was enhanced, and the TNF-alpha mediated decrease in eNOS mRNA was attenuated. Cerivastatin increased the stability of eNOS mRNA. However, it did not alter the stability of GTPCH mRNA but increased GTPCH gene transcription, as shown by nuclear run-on assays. Preteatment of HUVECs with the selective GTPCH inhibitor, 2,4-diamino-6 hydroxypyrimidine, caused a decrease in intracellular BH4 and decreased citrulline formation after stimulation with ionomycin. Furthermore, the potentiating effect of cerivastatin was decreased by limiting the cellular availability of BH4. CONCLUSIONS: Our data demonstrate that statins elevate GTPCH mRNA, thereby increasing BH4 levels in vascular endothelial cells. In addition to augmenting eNOS expression, statins potentiate GTPCH gene expression and BH4 synthesis, thereby increasing NO production and preventing relative shortages of BH4. PMID- 12588757 TI - Fluvastatin prevents renal dysfunction and vascular NO deficit in apolipoprotein E-deficient mice. AB - OBJECTIVE: The objective of this study was to investigate the effects of fluvastatin on atherosclerosis, systemic and regional hemodynamics, and vascular reactivity in apolipoprotein E-deficient (ApoE(-/-)) mice. METHODS AND RESULTS: Hemodynamics (fluospheres) and vasomotor responses of thoracic aorta and carotid artery were evaluated in male wild-type (WT) and untreated (ApoE(-/-) Control) or fluvastatin-treated (50 mg/kg per day for 20 weeks) ApoE(-/-) mice, all fed a Western-type diet. Plasma cholesterol and aortic root atherosclerotic lesions (ALs) were greater in ApoE(-/-) Control mice (19+/-1 mmol/L and 63,0176+/-38,785 micro m(2), respectively) than in WT mice (2+/-1 mmol/L and 1+/-1 micro m(2), respectively, P<0.01). Fluvastatin significantly decreased plasma cholesterol ( 53%) but failed to limit ALs. Renal blood flow was significantly reduced in ApoE( /-) Control versus WT (-25%, P<0.05) mice. This reduction was prevented by fluvastatin. Aortic and carotid endothelium-dependent relaxations to acetylcholine were not altered in ApoE(-/-) Control versus WT mice. In carotid arteries from WT mice, these responses were abolished after nitro-L-arginine (L NA), whereas those from ApoE(-/-) Control were only partially inhibited after L NA but fully abolished after L-NA+diclofenac. Thus, in carotid arteries from ApoE(-/-) mice, vasodilating prostanoids compensate the deficit in NO availability. Fluvastatin prevented this carotid NO deficit. CONCLUSIONS: In ApoE(-/-) mice, chronic fluvastatin treatment preserved renal perfusion and vascular NO availability independently from atherosclerotic lesion prevention. PMID- 12588758 TI - Distinct roles of ephrin-B2 forward and EphB4 reverse signaling in endothelial cells. AB - OBJECTIVE: The transmembrane ligand ephrin-B2 and its receptor tyrosine kinase EphB4 are specifically expressed on arterial and venous endothelial cells, respectively, and bidirectional signals mediated by both proteins play an important role in vascular development. However, how such bidirectional signals are required for cell-cell adhesion or repulsion remains unclear. METHODS AND RESULTS: Using a cell line and sorted primary endothelial cells, we show that ephrin-B2 forward signaling through the EphB4 receptor inhibits cell adhesion, whereas EphB4 reverse signaling by the transmembrane ephrin-B2 ligand does not. Cell migration is also inhibited on immobilized ephrin-B2-Fc but not on EphB4-Fc protein. CONCLUSIONS: Ephrin-B2 forward signaling and EphB4 reverse signaling differentially affect cell adhesion and migration between arterial and venous endothelial cells. PMID- 12588759 TI - Molecular basis of cell membrane estrogen receptor interaction with phosphatidylinositol 3-kinase in endothelial cells. AB - OBJECTIVE: Nontranscriptional signaling mechanisms mediate some of the biological effects of estrogen, such as the rapid actions on the blood vessels. By interacting with phosphatidylinositol 3-kinase (PI3K), estrogen receptor (ER) alpha leads to activation of protein kinase Akt and to subsequent increase in endothelial nitric oxide synthase activity. Because PI3K is mainly a cytoplasmic complex, we studied the cellular site of interaction between this enzyme and ERalpha, and we dissected the molecular mechanisms that mediate this interaction. METHODS AND RESULTS: By using cultured human saphenous vain endothelial cells, we found that cell membrane-bound ERalpha colocalizes with PI3K and may be responsible for PI3K activation. Furthermore, we characterized the subsequent steps in the activation of the PI3K/Akt signaling cascade, comparing the molecular events that follow insulin or estradiol activation of PI3K. CONCLUSIONS: We provide novel evidence for an important role of nonnuclear estrogen receptor in rapid, nontranscriptional responses of human endothelial cells to estrogen. PMID- 12588760 TI - Role of p44/p42 MAP kinase in the age-dependent increase in vascular smooth muscle cell proliferation and neointimal formation. AB - OBJECTIVE: Age-dependent increase in vascular smooth muscle cell (VSMC) proliferation is thought to contribute to the pathology of atherosclerotic diseases. In this study, we investigated the role of mitogen-activated protein kinases (MAPKs) on VSMC proliferation and neointimal formation in the context of aging. METHODS AND RESULTS: VSMCs were isolated from the aorta of young and old rabbits. The proliferative index after serum stimulation was significantly increased in old versus young VSMCs. This was associated with a significant and specific age-dependent increase in p44/p42 MAPK activation. Treatment with MEK inhibitor PD98059 successfully inhibited p44/p42 MAPK activities and VSMC proliferation. These results were confirmed in vivo using a model of balloon injury in rabbit iliac arteries. p44/p42 MAPK activities were rapidly induced by angioplasty in young and old animals. However, the levels of p44/p42 MAPK activities achieved in arteries of old rabbits were significantly higher than those of young rabbits. This was associated with a higher cellular proliferative index and a significant increase in neointimal formation in old animals. Local delivery of PD98059 in old rabbits successfully inhibited p44/p42 MAPK activities after angioplasty, which led to a significant reduction in cellular proliferation and neointimal formation in treated animals. CONCLUSIONS: Our study suggests for the first time that increased p44/p42 MAPK activation contributes to augmented VSMC proliferation and neointimal formation with aging. p44/p42 MAPK inhibition could represent a novel therapeutic avenue against atherosclerotic diseases. PMID- 12588761 TI - Involvement of RhoA/Rho kinase signaling in VEGF-induced endothelial cell migration and angiogenesis in vitro. AB - OBJECTIVE: Growth factor-induced angiogenesis involves migration of endothelial cells (ECs) into perivascular areas and requires active remodeling of the endothelial F-actin cytoskeleton. The small GTPase RhoA previously has been implicated in vascular endothelial growth factor (VEGF)-induced signaling pathways, but its role has not been clarified. METHODS AND RESULTS: VEGF induced the activation of RhoA and recruited RhoA to the cell membrane of human ECs. This increase in RhoA activity is necessary for the VEGF-induced reorganization of the F-actin cytoskeleton, as demonstrated by adenoviral transfection of dominant negative RhoA. Rho kinase mediated this effect of RhoA, as was demonstrated by the use of Y-27632, a specific inhibitor of Rho kinase. Inhibition of Rho kinase prevented the VEGF-enhanced EC migration in response to mechanical wounding but had no effect on basal EC migration. Furthermore, in an in vitro model for angiogenesis, inhibition of either RhoA or Rho kinase attenuated the VEGF mediated ingrowth of ECs in a 3-dimensional fibrin matrix. CONCLUSIONS: VEGF induced cytoskeletal changes in ECs require RhoA and Rho kinase, and activation of RhoA/Rho kinase signaling is involved in the VEGF-induced in vitro EC migration and angiogenesis. PMID- 12588762 TI - Lysophosphatidylcholine inhibits endothelial cell migration by increasing intracellular calcium and activating calpain. AB - OBJECTIVE: Endothelial cell (EC) migration, essential for reestablishing arterial integrity after vascular injury, is inhibited by oxidized LDL (oxLDL) and lysophosphatidylcholine (lysoPC) that are present in the arterial wall. We tested the hypothesis that a mechanism responsible for lysoPC-induced inhibition is increased intracellular free calcium concentration ([Ca(2+)](i)). METHODS AND RESULTS: LysoPC, at concentrations that inhibit in vitro EC migration to 35% of control, increased [Ca(2+)](i) levels 3-fold. These effects of lysoPC were concentration dependent and reversible. LysoPC induced Ca(2+) influx within 10 minutes, and [Ca(2+)](i) remained elevated for 2 hours. The calcium ionophore A23187 also increased [Ca(2+)](i) and inhibited EC migration. Chelators of intracellular Ca(2+) (BAPTA/AM and EGTA/AM) and nonvoltage-sensitive channel blockers (lanthanum chloride and gadolinium chloride) blunted the lysoPC-induced [Ca(2+)](i) rise and partially preserved EC migration. After lysoPC treatment, calpain, a calcium-dependent cysteine protease, was activated, and cytoskeletal changes occurred. Calpain inhibitors (calpastatin, MDL28170, and calpeptin) added before lysoPC prevented cytoskeletal protein cleavage and preserved EC migration at 60% of control levels. CONCLUSIONS: LysoPC increases [Ca(2+)](i). In turn, activating calpains that can alter the cytoskeleton are activated and EC migration is inhibited. PMID- 12588763 TI - Lysophosphatidic acid induction of tissue factor expression in aortic smooth muscle cells. AB - OBJECTIVE: Tissue factor (TF), the initiator of the coagulation cascade, is expressed by cells in atherosclerotic lesions. Lysophosphatidic acid (LPA) is a component of oxidized lipoproteins and an agent released by activated platelets. The present study investigated whether and how TF expression is regulated by LPA. METHODS AND RESULTS: Northern blotting, Western blotting, and TF activity assays demonstrated that LPA markedly induced TF mRNA, protein, and activity in vascular smooth muscle cells. LPA-induced TF expression is primarily controlled at the transcriptional level. Phosphorylation of mitogen-activated protein kinase kinase (MEK) and extracellular signaling-regulated kinases (ERK1/2) was rapidly and markedly induced by LPA. MEK inhibitors U0126 and PD98059 blocked both ERK activation and the increase in TF mRNA. In contrast, the specific p38 MAP kinase inhibitor SB203580 had no effect on LPA-induced TF mRNA increase. The Galpha(i) protein inhibitor, pertussis toxin, abolished LPA-induced phosphorylation of MEKs and ERKs, as well as the induction of TF mRNA. CONCLUSIONS: Our data demonstrate that a Galpha(i) protein and activation of MEKs and ERKs mediate LPA-induced TF expression. Our data suggest that elevated LPA could be a thrombogenic risk factor by upregulating TF expression. These results may have important implications in vascular remodeling and vascular diseases. PMID- 12588764 TI - HEX acts as a negative regulator of angiogenesis by modulating the expression of angiogenesis-related gene in endothelial cells in vitro. AB - OBJECTIVE: The hematopoietically expressed homeobox (HEX) is transiently expressed in endothelial cells (ECs) during vascular formation in embryo. Here, we investigated whether HEX played any role in angiogenesis-related properties of ECs in vitro. METHODS AND RESULTS: We transiently overexpressed HEX in human umbilical vein ECs (HUVECs). To our surprise, HEX completely abrogated the response of HUVECs to vascular endothelial growth factor (VEGF) with regard to proliferation, migration, and invasion and abolished network formation by HUVECs on Matrigel. cDNA microarray analysis and quantitative real-time reverse transcription-polymerase chain reaction combined with Western blotting revealed that HEX significantly repressed the expression of VEGF receptor-1, VEGF receptor 2, neuropilin-1, tyrosine kinase with Ig and EGF homology domains (TIE)-1, TIE-2, and the integrin alpha(v) subunit, whereas it augmented the expression of endoglin in HUVECs. We established murine embryonic stem cells that were stably transfected with HEX sense cDNA or antisense cDNA, and we examined the in vitro differentiation to ECs. Although the expression of VEGF receptor-2 was decreased in sense transfectants, the number of cells expressing VE-cadherin, a specific marker of ECs, was not altered. CONCLUSIONS: Our present results suggest that HEX may not affect the differentiation of ECs but acts as a negative regulator of angiogenesis. PMID- 12588765 TI - Mast cell chymase induces smooth muscle cell apoptosis by a mechanism involving fibronectin degradation and disruption of focal adhesions. AB - OBJECTIVE: Chymase released from activated mast cells has been shown to induce apoptosis of vascular smooth muscle cells (SMCs) in vitro. The proteolytic activity of chymase is essential for the proapoptotic effect, but the mechanism of chymase-induced apoptosis has remained unknown. METHODS AND RESULTS: Here we show by means of FACS analysis, immunohistochemistry, and Western blotting that mast cell-derived chymase induces SMC apoptosis by a mechanism involving degradation of an extracellular matrix component, fibronectin (FN), with subsequent disruption of focal adhesions. The FN degradation products induced SMC apoptosis of similar magnitude and with similar changes in outside-in signaling, as did chymase. Sodium orthovanadate, an inhibitor of tyrosine phosphatases, inhibited the chymase-induced SMC apoptosis. Focal adhesion kinase (FAK), one of the key mediators of integrin-extracellular matrix interactions and cell survival, was rapidly degraded in the presence of chymase or FN degradation products. Loss of phosphorylated FAK (p-FAK) resulted in a rapid dephosphorylation of the p-FAK-dependent downstream mediator Akt. CONCLUSIONS: The results suggest that chymase-secreting mast cells can mediate apoptosis of neighboring SMCs through a mechanism involving degradation of pericellular FN and disruption of the p-FAK-dependent cell-survival signaling cascade. PMID- 12588766 TI - Disparity of MCP-1 mRNA and protein expressions between the carotid artery and the aorta in WHHL rabbits: one aspect involved in the regional difference in atherosclerosis. AB - OBJECTIVE: This study was designed to examine why in WHHL rabbits, muscular arteries, such as the carotid artery, are relatively resistant to atherosclerosis compared with the aorta, with a special reference to monocyte chemoattractant protein (MCP)-1. METHODS AND RESULTS: MCP-1 mRNA expression was quantitated by Northern blot analysis, and its protein expression was quantitated by immunostaining and ELISA at the age of 1, 3, 6, and 12 months (n=5 to 6 each). In the aorta, atherosclerotic lesions were progressively developed with aging, and MCP-1 was highly expressed in endothelial cells and infiltrating macrophages. By contrast, in the carotid artery, atherosclerotic lesions and MCP-1 immunoreactivity were not evident throughout the experimental period. Unexpectedly, however, the extent of MCP-1 mRNA expression was comparable between the aorta and the carotid artery throughout the experimental period. Endothelial cells in primary culture from the aorta and the carotid artery expressed the same extent of MCP-1 mRNA on stimulation by oxidized LDL. There was no abnormality in primary structure of MCP-1 cDNA in WHHL. CONCLUSIONS: These results suggest that in WHHL, the atherosclerosis process, including MCP-1 protein expression, may be reduced in the carotid artery (and possibly in other muscular arteries), accounting in part for the regional resistance to atherosclerosis. PMID- 12588767 TI - ACE-versus chymase-dependent angiotensin II generation in human coronary arteries: a matter of efficiency? AB - OBJECTIVE: The objective of this study was to investigate ACE- and chymase dependent angiotensin I-to-II conversion in human coronary arteries (HCAs). METHODS AND RESULTS: HCA rings were mounted in organ baths, and concentration response curves to angiotensin II, angiotensin I, and the chymase-specific substrate Pro(11)-D-Ala(12)-angiotensin I (PA-angiotensin I) were constructed. All angiotensins displayed similar efficacy. For a given vasoconstriction, bath (but not interstitial) angiotensin II during angiotensin I and PA-angiotensin I was lower than during angiotensin II, indicating that interstitial (and not bath) angiotensin II determines vasoconstriction. PA-angiotensin I increased interstitial angiotensin II less efficiently than angiotensin I. Separate inhibition of ACE (with captopril) and chymase (with C41 or chymostatin) shifted the angiotensin I concentration-response curve approximately 5-fold to the right, whereas a 10-fold shift occurred during combined ACE and chymase inhibition. Chymostatin, but not captopril and/or C41, reduced bath angiotensin II and abolished PA-Ang I-induced vasoconstriction. Perfused HCA segments, exposed luminally or adventitially to angiotensin I, released angiotensin II into the luminal and adventitial fluid, respectively, and this release was blocked by chymostatin. CONCLUSIONS: Both ACE and chymase contribute to the generation of functionally active angiotensin II in HCAs. However, because angiotensin II loss in the organ bath is chymase-dependent, ACE-mediated conversion occurs more efficiently (ie, closer to AT(1) receptors) than chymase-mediated conversion. PMID- 12588768 TI - Expansive remodeling is a response of the plaque-related vessel wall in aortic roots of apoE-deficient mice: an experiment of nature. AB - OBJECTIVE: In the present study, we (1) evaluated the apolipoprotein E-deficient (apoE(-/-)) mouse aortic root as a model for expansive remodeling in atherosclerosis and (2) examined whether remodeling at this site was due to dilation of the vessel wall beneath or flanking the plaque. METHODS AND RESULTS: Plaque area and length of the internal elastic lamina (IEL) were measured in aortic roots of apoE(-/-) mice at 11 weeks, 6 months, and 13 months of age. Expansive remodeling, indicated by IEL lengthening exceeding that of normal growth in wild-type C57BL/6 mice, was evident at 6 months of age and showed overcompensation (luminal enlargement) by 13 months of age. When the left coronary (LC), right coronary (RC), and noncoronary (NC) sinuses of the aortic root were looked at individually, remodeling was located to the LC at 6 months of age and to the LC and RC at 13 months of age. The remodeling response was closely related to local plaque area but unrelated to plaque formation in flanking sinuses. CONCLUSIONS: The aortic root of the apoE(-/-) mouse features a consistent remodeling response. In this model, expansive remodeling was a strictly local response of the plaque-related vessel wall. PMID- 12588769 TI - Macrophage-expressed group IIA secretory phospholipase A2 increases atherosclerotic lesion formation in LDL receptor-deficient mice. AB - OBJECTIVE: Transgenic mice expressing human group IIA secretory phospholipase A(2) (group IIA sPLA(2)) spontaneously develop atherosclerotic lesions. The mechanism for this proatherogenic effect is likely multifactorial, because HDL cholesterol is significantly lower and LDL/VLDL cholesterol is slightly higher in transgenic mice compared with nontransgenic littermates. In the present study, we show for the first time that elicited peritoneal macrophages from transgenic mice express human group IIA sPLA(2). This study tested whether macrophage-expressed sPLA(2) contributes to atherogenesis. METHODS AND RESULTS: Bone marrow cells from either sPLA(2) transgenic mice or control C57BL/6 mice were transplanted into LDL receptor-deficient mice. After hematopoietic engraftment, animals were fed a diet enriched with saturated fat and cholesterol for 12 weeks. Despite a lack of effect on serum lipoprotein concentrations, the presence of bone marrow-derived cells expressing human group IIA sPLA(2) resulted in a significant increase in the extent of atherosclerosis in the aortic arch (12.8+/-1.4% versus 7.4+/-0.9%; P<0.005) and aortic sinus (0.3+/-0.03 mm(2) versus 0.2+/-0.04 mm(2); P<0.05). CONCLUSIONS: Group IIA sPLA(2) can contribute to atherosclerotic lesion development through a mechanism that is independent of systemic lipoprotein metabolism. PMID- 12588770 TI - Isoform-dependent cholesterol efflux from macrophages by apolipoprotein E is modulated by cell surface proteoglycans. AB - OBJECTIVE: Apolipoprotein E (apoE) mediates cellular cholesterol efflux and plays a crucial role in the inhibition of atherogenesis. We investigated whether there is an isoform-specific difference in its function for cholesterol efflux from cholesterol-loaded RAW264.7 cells, a murine macrophage cell line that lacks endogenous apoE expression. METHODS AND RESULTS: When human apoE was expressed in RAW264.7 cells, apoE2 reduced cellular total cholesterol (TC) and esterified cholesterol (EC) levels significantly, whereas apoE3 and apoE4 had no effect. However, treatment of cells with 4-methylumbelliferyl-7-beta-D-xyloside (beta-DX) resulted in all 3 isoforms' reducing cellular TC and EC contents significantly. We also investigated the effect of exogenously derived apoE on cholesterol efflux by utilizing the medium harvested from HeLa cells expressing apoE. ApoE2 and E3 reduced both cellular TC and EC contents significantly, whereas apoE4 did not. However, treatment of the cells with beta-DX resulted in all 3 exogenously derived apoE isoforms' reducing TC and EC contents significantly. The binding ability of apoE to heparan sulfate proteoglycans examined by heparinase I treatment revealed less binding ability of apoE2 compared with that of apoE3 or apoE4. CONCLUSIONS: The present study clarified the differential cellular cholesterol-modulating effect of apoE isoforms in macrophages, which would be due to the difference in their binding to proteoglycans. PMID- 12588771 TI - Phospholipid hydroxyalkenals: biological and chemical properties of specific oxidized lipids present in atherosclerotic lesions. AB - OBJECTIVE: Phosphatidylcholine hydroxyalkenals (PC-HAs) are a class of oxidized PCs derived from lipid peroxidation of arachidonate or linoleate at the sn-2 position to form terminal gamma-hydroxy, alpha-, and beta-unsaturated aldehydes. The aim of this study was to characterize some of their biological properties, ascertain the mechanism of their action, and assess whether they have in vivo relevance. METHODS AND RESULTS: Combinations of cell biological approaches with radiolabels, mass spectroscopy, and immunochemical as well as immunohistochemical techniques were used to show that PC-HAs reduce the proteolytic degradation by mouse peritoneal macrophages (MPMs) of internalized macromolecules, such as maleylated bovine serum albumin, and that the activity of the lysosomal protease, cathepsin B, in MPMs form Michael adducts with MPM proteins and with N-acetylated cysteine in vitro form pyrrole adducts with MPM proteins and reduce the maturation of Rab5a, thereby impairing phagosome-lysosome fusion (maturation) in phagocytes; they are present unbound and as pyrrole adducts in human atherosclerotic lesions. CONCLUSIONS: PC-HAs are present in vivo and possess multiple functions characteristic of oxidized LDL and 4-hydroxynonenal. PMID- 12588772 TI - Antidiabetic PPAR gamma-activator rosiglitazone reduces MMP-9 serum levels in type 2 diabetic patients with coronary artery disease. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are critically involved in the development of unstable plaques. Although arteriosclerotic lesions in patients with diabetes mellitus are more unstable than those of nondiabetic subjects, nothing is known about serum levels of MMPs in these patients or about mechanisms to modulate MMP levels. We investigated MMP levels in diabetic and nondiabetic coronary artery disease (CAD) patients and performed a clinical trial to assess the effect of the PPARgamma-activating, antidiabetic thiazolidinedione rosiglitazone on MMP levels in diabetic CAD patients. METHODS AND RESULTS: In CAD patients, MMP-2, -8, and -9 serum levels were significantly higher in type 2 diabetic subjects compared with age-, sex-, and body mass index-matched nondiabetics. Thirty-nine diabetic patients with CAD were randomized to receive rosiglitazone 4 mg (twice daily) or placebo for 12 weeks. Rosiglitazone treatment, but not placebo, significantly reduced MMP-9 levels already after 2 weeks by -19.6% (-38.3% to 8.6%, P<0.05), and levels remained suppressed until the end of the study. In addition, rosiglitazone significantly decreased serum amyloid A (SAA) and tumor necrosis factor-alpha levels. CONCLUSION: MMP-9 levels are increased in type 2 diabetic patients with CAD, and treatment of these patients with the antidiabetic PPARgamma-activator rosiglitazone significantly reduces MMP-9, tumor necrosis factor-alpha, and SAA serum levels. These data support anti-inflammatory and potential antiatherogenic effects of thiazolidinediones. PMID- 12588773 TI - A functional polymorphism in a STAT5B site of the human PPAR gamma 3 gene promoter affects height and lipid metabolism in a French population. AB - OBJECTIVE: The peroxisome proliferator-activated receptor-gamma (PPARgamma) plays a role in adipocyte differentiation and insulin sensitization. It has been shown that genetic variation in the PPARgamma gene alters body weight control, lipid and insulin homeostasis, and the susceptibility to type 2 diabetes. Four PPARgamma isoforms are generated by alternative splicing and promoter usage. PPARgamma3 is only expressed in adipose tissue, colon, and macrophages and therefore seems to be a good candidate gene for metabolic and cardiovascular associated diseases. In the present study, we looked for genetic variation in the PPARgamma3 promoter. METHODS AND RESULTS: The proximal PPARgamma3 promoter was sequenced in 20 individuals. We detected a C/G polymorphism at position -681 from exon A2. Interestingly, it was located in a signal transducer and activator of transcription 5B (STAT5B) binding consensus site. In a French population (n=836), the -681G allele was associated with increased height and plasma low-density lipoprotein cholesterol concentrations. In vitro, we showed that the -681G allele completely abolished the binding of STAT5B to the cognate promoter element as well as the transactivation of the PPARgamma3 promoter by the growth hormone/STAT5B pathway. CONCLUSIONS: Our results suggest that PPARgamma3 may regulate the control of height and lipid homeostasis via the STAT5B pathway. PMID- 12588774 TI - Association between insulin resistance and carotid arteriosclerosis in subjects with normal fasting glucose and normal glucose tolerance. AB - OBJECTIVE: We examined the possible association between insulin resistance and carotid arteriosclerosis in subjects who had both normal fasting glucose and normal glucose tolerance after intake of a glucose load. METHODS AND RESULTS: Our subjects were individuals who underwent general health screening at our institute, which included carotid ultrasound and oral glucose tolerance testing. Of the 1238 subjects enrolled in our study, 738 (60%) were classified as normal, defined as a normal fasting glucose level and normal glucose tolerance, and 334 (27%) and 166 (13%) were classified as borderline and diabetic, respectively, according to the criteria of the Japan Diabetes Society. The homeostasis model assessment of insulin resistance (HOMA-IR) was used as the index to measure insulin resistance. In normal-type subjects, univariate analysis showed that insulin resistance, but not insulin secretion, was associated with the presence of carotid plaque. Multivariate analysis showed that HOMA-IR was positively associated with carotid plaque in normal-type subjects, with an odds ratio of 1.19 (95% confidence interval, 1.00 to 1.41; P<0.05). CONCLUSIONS: These data suggest the possibility that the presence of higher insulin resistance could be a risk factor for carotid arteriosclerosis in subjects with normal fasting glucose and normal glucose tolerance. PMID- 12588775 TI - Early carotid atherosclerosis and family history of vascular disease: specific effects on arterial sites have implications for genetic studies. AB - OBJECTIVE: Carotid artery intima-media thickness (IMT) is an intermediate phenotype for atherosclerosis. In a community population (n=5400), we determined whether a parental history of myocardial infarction (MI) or stroke is associated with increased IMT and whether associations differ at specific sites in the carotid arterial tree. METHODS AND RESULTS: Using regression modeling, the proportion of IMT that remains unexplained after controlling for vascular risk factors was determined. A parental history of stroke was associated with both increased common carotid artery (CCA) and increased internal carotid artery (ICA) IMT, but in young individuals (or=5.2%) than in the first quartile (<2.5%, P=0.03). Stepwise multiple regression analysis showed that LDL-IVb was the single best predictor of stenosis change. CONCLUSIONS: LDL-IVb was the single best lipoprotein predictor of increased stenosis, an unexpected result, given that LDL-IVb represents only a minor fraction of total LDL. PMID- 12588778 TI - Treatment with statins after induction of focal ischemia in rats reduces the extent of brain damage. AB - OBJECTIVE: In the present study, MRI has been used to investigate therapeutic intervention with statins in a model of permanent focal cerebral ischemia in rat. METHODS AND RESULTS: Brain ischemia was induced in rats by the permanent occlusion of middle cerebral artery (MCAO) and the brain infarct size followed up in alive animals 2, 24, and 48 hours after MCAO, using the trace of apparent diffusion coefficient [Tr(D)] maps and T2-weighted images. In vehicle-treated rats, the infarct volumes increased by 38.5% and 89% after 24 and 48 hours, respectively, compared with the damage detected at 2 hours after MCAO. Treatment with simvastatin (20 mg/kg) after MCAO prevented the increase in brain infarct volume occurring at 24 hours and induced a 46.6% reduction after 48 hours. This effect was similar to that observed when simvastatin was administered before the induction of focal ischemia. T2W-MRI images confirmed these findings. The neuroprotective effects of simvastatin were paralleled by an increase in endothelial NO synthase immunoreactivity, detectable in the brain of simvastatin treated rats. CONCLUSIONS: Statins, in addition to their preventive effect on cerebral ischemia, exert a neuroprotective role in the attenuation of brain damage after acute stroke. PMID- 12588779 TI - Extensive association analysis between polymorphisms of PON gene cluster with coronary heart disease in Chinese Han population. AB - OBJECTIVE: An extensive association analysis of PON gene cluster (PONs) with coronary heart disease (CHD) was performed in Chinese Han population. METHODS AND RESULTS: Thirty polymorphisms of PON1, PON2, and PON3 gene were identified by direct sequencing of genomic DNA derived from 48 randomly selected patients. Twelve polymorphisms were additionally investigated for association with CHD in 474 male patients and 475 controls. Univariate analyses showed the cases had significantly higher frequencies of PON1 192Q allele, 160R allele, -162A allele, and PON2 311C allele than were seen in the controls. Logistic regression analyses revealed only the PON1 R160G and -162G/A polymorphisms remained significantly associated with CHD (P=0.0054 and P=0.0002). Haplotype analyses for various polymorphism combinations additionally confirmed the results of individual polymorphism analyses. Only the frequencies of haplotypes containing -162A allele were significantly higher, whereas only the frequencies of haplotypes containing 160G allele were significantly lower in cases than in controls in various polymorphism combinations. CONCLUSIONS: This extensive association study has identified the PON1 -162G/A and R160G polymorphisms to be independently associated with CHD in Chinese Han population and warrants additional study to elucidate the biological mechanism. PMID- 12588780 TI - Systematic family screening for familial hypercholesterolemia in Iceland. AB - OBJECTIVE: This study compares a novel approach using systematic family screening for patients in Iceland who have familial hypercholesterolemia (FH) with conventional proband screening and assesses the sensitivity and specificity of diagnosing FH by cholesterol measurements compared with mutational testing of family members. METHODS AND RESULTS: Probands with the I4T+2C mutation were traced to common ancestors. A downtracing of each family lineage was performed back to the oldest living offspring (key individuals); these individuals were recruited for cholesterol measurement and mutation testing. The sensitivity and specificity of cholesterol measurements was assessed against mutational analysis. Eleven probands clustered into 4 families. There were 364 key individuals identified among their descendants. Eighty-four percent responded, and 11% were positive for the mutation. There were 78 offspring of the positive key individuals, and 40 of those were carriers. Compared with use of the conventional first-degree relative approach, an additional 19% of FH individuals, including key individuals and their descendants, were identified. As diagnostic criteria, cholesterol measurements in the families had 95% specificity and 94% sensitivity. CONCLUSIONS: Tracing FH probands to common ancestors and screening the oldest offspring in each family lineage adds considerably to the conventional method of FH screening (testing first-degree relatives). This may have relevance in other founder populations. PMID- 12588781 TI - A quantitative trait locus on chromosome 16q influences variation in plasma HDL-C levels in Mexican Americans. AB - OBJECTIVE: We conducted a whole-genome, multipoint linkage screen to localize a previously reported major locus accounting for 56% to 67% of the additive genetic effects on covariate-adjusted plasma HDL cholesterol (HDL-C) levels in Mexican Americans from the San Antonio Family Heart Study (SAFHS). METHODS AND RESULTS: After using complex segregation analysis to recover the major locus in 472 SAFHS participants from 10 genotyped families, we incorporated covariates required to detect that major locus, including plasma levels of triglycerides and apolipoprotein A-I, in a maximum-likelihood-based variance-components linkage screen. Only chromosome 16 exhibited convincing evidence for a quantitative trait locus (QTL), with a peak multipoint log of the odds (LOD)=3.73 (P=0.000034). Subsequent penetrance model-based linkage analysis, incorporating genotypes at the marker locus nearest the multipoint peak (D16S518) into the segregation model, detected linkage with the previously detected major locus (LOD=2.73, P=0.000642). Initial estimates place this QTL within a 15-cM region of chromosome 16q near the structural loci for lecithin:cholesterol acyltransferase (LCAT) and cholesteryl ester transfer protein (CETP). CONCLUSIONS: A QTL influencing plasma levels of HDL-C in Mexican Americans from San Antonio maps to a region of human chromosome 16q near LCAT and CETP. PMID- 12588782 TI - A novel nonobstructive intravascular MRI coil: in vivo imaging of experimental atherosclerosis. AB - OBJECTIVE: MRI is being used to characterize the composition of atherosclerotic plaques. However, the resolution achievable using surface radiofrequency coils is limited by the signal to noise ratio. We studied the utility of a new intravascular (IV) MRI probe for high-resolution in vivo imaging of atherosclerotic lesions. METHODS AND RESULTS: Balloon-injured Watanabe heritable hyperlipidemic rabbits served as the experimental model of atherosclerosis. The newly developed IV MRI probe is 1.3 mm in diameter and can be positioned over a guidewire. MRI was performed with both an external phased-array coil and the IV MR coil. MR observations were correlated with histopathology. After MRI, the animals were killed and analysis of agreement between MR and histopathology was performed. The IV MR coil allows aortic images to be obtained with 156x156 micro m(2) in-plane resolution versus 352x352 micro m(2) when used with the external phased-array coil. No significant motion artifacts were noted, despite the continuation of arterial blood flow during image acquisition around the IV probe. The different components of the atherosclerotic lesions (lipid core and fibrous cap) were easily identified. There was an excellent agreement between MRI with the IV coil and histopathology by simple linear regression for both the mean wall thickness (r=0.88, slope 0.82, P<0.0001) and vessel wall area (r=0.86, slope 1.08, P<0.0001). CONCLUSIONS: The new nonobstructive design for this intravascular coil provides great promise for additional work in high-resolution MRI characterization of atherosclerotic plaques in vivo. The ability to position the probe with a guidewire allows its placement under fluoroscopic or MRI guidance, whereas its size is compatible with human coronary arteries. PMID- 12588783 TI - Bicuspid aortic valve associated with aortic dilatation: a community-based study. AB - OBJECTIVE: This study was undertaken to examine the association between bicuspid aortic valve (BAV) and aortic dilatation in the community. The association between BAV and aortic dilatation has been reported predominantly in retrospective studies in patients mostly with valvular dysfunction or selected surgical patients from tertiary referral centers. An independent association of BAV and aortic dilatation in a community-based study has not been demonstrated. METHODS AND RESULTS: In a geographically defined population of Olmsted County, Minnesota, residents with BAV (n=44, age 35+/-13 years) without hemodynamically significant obstruction or regurgitation and matched controls with normal tricuspid aortic valves were identified by transthoracic echocardiography. The two groups were compared with respect to measurements of the aorta. The BAV and control groups differed with respect to size of the aortic anulus (23.2+/-2.4 versus 21.6+/-2.4 mm; P=0.002), aortic sinus (33.5+/-4.6 versus 30.3+/-4.1 mm; P=0.0001), and proximal ascending aorta (33.3+/-6.5 versus 27.9+/-3.6 mm; P=0.0001). There was no difference in the size of the aortic arch (24.2+/-3.6 versus 25.3+/-3.4 mm; P=0.16). These differences were maintained when the groups were stratified by sex and blood pressure. The relationship between bicuspid aortic valve and aortic dilatation was maintained when adjusting for factors related to fluid mechanics and hemodynamics such as systolic blood pressure, diastolic blood pressure, left ventricular ejection time, and peak aortic valve velocity. CONCLUSIONS: In a community-based study, BAV is associated with an alteration of aortic dimensions even in the absence of hemodynamically significant aortic valve stenosis or regurgitation. PMID- 12588784 TI - Blockade of the platelet P2Y12 receptor by AR-C69931MX sustains coronary artery recanalization and improves the myocardial tissue perfusion in a canine thrombosis model. AB - OBJECTIVE: Reperfusion therapy for myocardial infarction is limited by a significant reocclusion rate and less optimal myocardial tissue perfusion due to excessive platelet accumulation and recruitment at the sites of vascular injury. We assessed the influence of a selective P2Y(12)-receptor antagonist (AR C69931MX), in conjunction with thrombolytic therapy, on the prevention of platelet aggregation and thrombus formation. METHODS AND RESULTS: A canine coronary electrolytic injury thrombosis model was used. Tissue-type plasminogen activator (t-PA; 1 mg/kg in phase I, 0.5 mg/kg in phase II in the AR-C69931MX group, and 1 mg/kg in the placebo group in phase I and II) was administered 30 minutes after thrombus formation; either saline or AR-C69931MX (4 micro g x kg( 1) x min(-1)) was given to all animals intravenously 10 minutes before t-PA administration for a total of 2 hours. All animals received heparin (80 U/kg) as an intravenous bolus followed by a continuous infusion of 17 U x kg(-1) x h(-1). Myocardial tissue perfusion was evaluated by use of the colored microsphere technique and real-time myocardial contrast echocardiography. The incidences of reocclusion and cyclic flow variation were significantly decreased in the AR C69931MX group (P<0.05). Myocardial tissue flow with AR-C69931MX treatment improved significantly at 20 and 120 minutes after reflow, whereas tissue flow with placebo remained at a level similar to that during occlusion (P<0.05). CONCLUSIONS: The adjunctive administration of AR-C69931MX blocked ADP-mediated platelet aggregation and recruitment and prevented platelet-mediated thrombosis, resulting in prolongation of reperfusion time and a decrease in reocclusion and cyclic flow variations. Importantly, myocardial tissue perfusion was significantly improved in the P2Y(12) antagonist group. PMID- 12588785 TI - Fish consumption, fish oil, omega-3 fatty acids, and cardiovascular disease. PMID- 12588786 TI - ATP consumption rate per cross bridge depends on myosin heavy chain isoform. AB - In the present study, we tested the hypothesis that intrinsic differences in ATP consumption rate per cross bridge exist across rat diaphragm muscle (Dia(m)) fibers expressing different myosin heavy chain (MHC) isoforms. During maximum Ca(2+) activation (pCa 4.0) of single, Triton X-permeabilized Dia(m) fibers, isometric ATP consumption rate was determined by using an NADH-linked fluorometric technique. The MHC concentration in single Dia(m) fibers was determined by densitometric analysis of SDS-PAGE gels and comparison to a standard curve of known MHC concentrations. Isometric ATP consumption rate varied across Dia(m) fibers expressing different MHC isoforms, being highest in fibers expressing MHC(2X) (1.14 +/- 0.08 nmol. mm(-3). s(-1)) and/or MHC(2B) (1.33 +/- 0.08 nmol. mm(-3). s(-1)), followed by fibers expressing MHC(2A) (0.77 +/- 0.11 nmol. mm(-3). s(-1)) and MHC(Slow) (0.46 +/- 0.03 nmol. mm(-3). s(-1)). These differences in ATP consumption rate also persisted when it was normalized for MHC concentration in single Dia(m) fibers. Normalized ATP consumption rate for MHC concentration varied across Dia(m) fibers expressing different MHC isoforms, being highest in fibers expressing MHC(2X) (2.02 +/- 0.19 s(-1)) and/or MHC(2B) (2.64 +/- 0.15 s(-1)), followed by fibers expressing MHC(2A) (1.57 +/- 0.16 s( 1)) and MHC(Slow) (0.77 +/- 0.05 s(-1)). On the basis of these results, we conclude that there are intrinsic differences in ATP consumption rate per cross bridge in Dia(m) fibers expressing MHC isoforms. PMID- 12588787 TI - CT-based assessment of regional pulmonary microvascular blood flow parameters. AB - To determine regional pulmonary microvascular mean transit times (MTTs), we used electrocardiogram-gated X-ray computed tomographic imaging to follow bolus radiopaque contrast material through the lungs in anesthetized animals (7 dogs and 1 pig, prone and supine). By deconvolution/reconvolution of regional time attenuation curves obtained from parenchyma and large lobar arteries, we estimated the microvascular residue function and reconstituted the regional microvascular time-attenuation curves and, thus, regional microvascular MTTs. The mean microvascular MTTs in the supine and prone postures were 3.94 +/- 1.0 and 3.40 +/- 0.84 (mean +/- SD), respectively. The dependent-nondependent vertical gradient of MTT was greater in the supine [slope = 0.25 +/- 0.10 (SD), P < 0.001 by t-test] than in the prone (-0.03 +/- 0.06 in 6 of 8 animals; 2 outliers had positive slopes) posture. In both postures, there was a trend toward faster transit times in the dorsal-basal lung region in six of the eight animals, suggesting gravity-independent higher vascular conductance dorsocaudally. We conclude that deconvolution methods, in association with electrocardiogram-gated high-speed X-ray computed tomography, can provide insights into regional heterogeneity of pulmonary microvascular MTT in vivo. PMID- 12588788 TI - Effect of exogenous growth hormone and exercise on lean mass and muscle function in children with burns. AB - We tested the hypothesis that the administration of recombinant human growth hormone (rHGH) and exercise would increase lean body mass (LBM) and muscle strength in burned children to a greater extent than rHGH or exercise separately. Children, ages 7-17 yr, with >40% body surface area burned, were randomized into groups. One group (GHEX, n = 10) participated in a 12-wk in-hospital physical rehabilitation program supplemented with an exercise program and received 0.05 mg. kg(-1). day(-1) of rHGH. A second exercising group (SALEX, n = 13) received saline. A third group (GH, n = 10) received a similar dose of rHGH as GHEX and participated in a 12-wk, home-based physical rehabilitation program without exercise. The fourth group (Saline, n = 11) received saline and participated in a 12-wk, home-based physical rehabilitation program without exercise. The mean (+/ SE) percent change in lean body mass after 12 wk was not significantly different between GHEX (9.0 +/- 2.1%), SALEX (5.4 +/- 1.6%), and GH (5.8 +/- 1.8%) groups (P = 0.33). However, the mean percent change in muscle strength was significantly greater in the GHEX (36.2 +/- 5.4%) and SALEX (42.6 +/- 10.0%) groups than in the GH (-7.4 +/- 4.7%) or Saline (6.7 +/- 4.4%) groups (P = 0.008). In summary, rHGH GHEX, SALEX, and GH alone produced similar improvements in LBM. However, muscle strength was only increased via exercise. PMID- 12588789 TI - Facilitation of triceps brachii muscle contraction by tendon vibration after chronic cervical spinal cord injury. AB - One way to improve the weak triceps brachii voluntary forces of people with chronic cervical spinal cord injury may be to excite the paralyzed or submaximally activated fraction of muscle. Here we examined whether elbow extensor force was enhanced by vibration (80 Hz) of the triceps or biceps brachii tendons at rest and during maximum isometric voluntary contractions (MVCs) of the elbow extensors performed by spinal cord-injured subjects. The mean +/- SE elbow extensor MVC force was 22 +/- 17.5 N (range: 0-23% control force, n = 11 muscles). Supramaximal radial nerve stimuli delivered during elbow extensor MVCs evoked force in six muscles that could be stimulated selectively, suggesting potential for force improvement. Biceps vibration at rest always evoked a tonic vibration reflex in biceps, but extension force did not improve with biceps vibration during triceps MVCs. Triceps vibration induced a tonic vibration reflex at rest in one-half of the triceps muscles tested. Elbow extensor MVC force (when >1% of control force) was enhanced by vibration of the triceps tendon in one-half of the muscles. Thus triceps, but not biceps, brachii tendon vibration increases the contraction strength of some partially paralyzed triceps brachii muscles. PMID- 12588790 TI - Physical exercise increases urinary excretion of lipoxin A4 and related compounds. AB - Lipoxins (LX) are lipoxygenase-derived eicosanoids with potent anti-inflammatory activities and vascular bed-dependent vasodilatory actions. LX can be formed in vitro and in vivo in a number of conditions, and we have reported that immunoreactive LXA(4) (iLXA(4)) is physiologically excreted with human urine. Using a recently developed LX extraction method coupled to an ELISA, we examined whether iLXA(4) excretion was modified by strenuous exercise, which is known to trigger potential LX-forming events. Maximal exertion significantly increased iLXA(4) urinary excretion in nine healthy volunteers (0.061 +/- 0.023 vs. 0.113 +/- 0.057 ng/mg creatinine; P = 0.028). iLXA(4) levels returned to baseline after 6 h and increased, although at a smaller extent, after 24 h. A significant correlation (r = 0.988) was denoted between iLXA(4) ELISA measurements and reversed-phase high-performance liquid chromatography quantitation of a previously described urinary tetraene, confirming its LXA(4)-related nature. These findings show for the first time that an increase in excretion of LXA(4) related compounds can be observed in response to strenuous exercise. This may be the reflection of an enhanced LX biosynthesis, which may represent a safeguard mechanism that keeps the inflammatory reaction triggered by physical stress under control. PMID- 12588791 TI - Intracellular sodium in mammalian muscle fibers after eccentric contractions. AB - The effect of eccentric contractions on intracellular Na(+) concentration ([Na(+)](i)) and its distribution were examined in isolated rat and mouse muscle fiber bundles. [Na(+)](i) was measured with either Na(+)-binding benzofuran isophthalate or sodium green. Ten isometric contractions had no significant effect on force (measured after 5 min of recovery) and caused no significant change in the resting [Na(+)](i) (7.2 +/- 0.5 mM). In contrast 10 eccentric contractions (40% stretch at 4 muscle lengths/s) reduced developed force at 100 Hz to 45 +/- 3% of control and increased [Na(+)](i) to 16.3 +/- 1.6 mM (n = 6; P < 0.001). The rise of [Na(+)](i) occurred over 1-2 min and showed only minimal recovery after 30 min. Confocal images of the distribution of [Na(+)](i) showed a spatially uniform distribution both at rest and after eccentric contractions. Gd(3+) (20 microM) had no effect on resting [Na(+)](i) or control tetanic force but prevented the rise of [Na(+)](i) and reduced the force deficit after eccentric damage. These data suggest that Na(+) entry after eccentric contractions may occur principally through stretch-sensitive channels. PMID- 12588792 TI - Effects of the nasal valve on acoustic rhinometry measurements: a model study. AB - The influence of nasal valve on acoustic rhinometry (AR) measurements was evaluated by using simple nasal cavity models. Each model consisted of a cylindrical pipe with an insert simulating the nasal valve. The AR-determined cross-sectional areas beyond the insert were consistently underestimated, and the corresponding area-distance curves showed pronounced oscillations. The area underestimation was more pronounced in models with inserts of small passage area. The experimental results are discussed in terms of theoretically calculated "sound-power reflection coefficients" for the pipe models. The reason for area underestimation is reflection of most of the incident sound power from the barrier at the front junction between the pipe and the insert. It was also demonstrated that the oscillations are due to low-frequency acoustic resonances in the portion of the pipe beyond the insert. The results suggest that AR does not provide reliable information about the cross-sectional areas of the nasal cavity posterior to a significant constriction, such as pathologies narrowing the nasal valve area. When the passage area of the nasal valve is decreased, the role of AR as a diagnostic tool for the entire nasal cavity becomes limited. PMID- 12588793 TI - Mutations in a new scaffold protein Sans cause deafness in Jackson shaker mice. AB - The Jackson shaker (js) mouse carries a recessive mutation causing phenotypes such as deafness, abnormal behavior (circling and/or head-tossing) and degeneration of inner ear neuroepithelia. Two alleles have been identified so far, the original js and js(seal). A contig of three BAC clones was isolated by positional cloning. Two of the clones rescue the js phenotype by BAC transgenesis. Analysis of transcripts in an overlapping region of the two clones revealed a gene encoding a new scaffold-like protein, Sans, that showed mutations in the two js mutants. One was a guanine nucleotide insertion in the original js allele and the other a 7-base insertion in the js(seal) allele. Both insertions are predicted to inactivate the Sans protein by frameshift mutations resulting in a truncated protein lacking the C-terminal SAM domain. Cochlear hair cells in the js mutants show disorganized stereocilia bundles, and Sans were highly expressed in inner and outer hair cells of cochlea. The existence of major motifs, ankyrin repeats and a SAM domain suggests that Sans may have an important role in the development and maintenance of the stereocilia bundles through protein-protein interaction. PMID- 12588794 TI - Usher syndrome type I G (USH1G) is caused by mutations in the gene encoding SANS, a protein that associates with the USH1C protein, harmonin. AB - Usher syndrome type I (USH1) is the most frequent cause of hereditary deaf blindness in humans. Seven genetic loci (USH1A-G) have been implicated in this disease to date, and four of the corresponding genes have been identified: USH1B, C, D and F. We carried out fine mapping of USH1G (chromosome 17q24-25), restricting the location of this gene to an interval of 2.6 Mb and then screened genes present within this interval for mutations. The genes screened included the orthologue of the Sans gene, which is defective in the Jackson shaker deaf mutant and maps to the syntenic region in mice. In two consanguineous USH1G-affected families, we detected two different frameshift mutations in the SANS gene. Two brothers from a German family affected with USH1G were found to be compound heterozygotes for a frameshift and a missense mutation. These results demonstrate that SANS underlies USH1G. The SANS protein contains three ankyrin domains and a sterile alpha motif, and its C-terminal tripeptide presents a class I PDZ-binding motif. We showed, by means of co-transfection experiments, that SANS associates with harmonin, a PDZ domain-containing protein responsible for USH1C. In Jackson shaker mice the hair bundles, the mechanoreceptive structures of inner ear sensory cells, are disorganized. Based on the known interaction between USH1B (myosin VIIa), USH1C (harmonin) and USH1D (cadherin 23) proteins and the results obtained in this study, we suggest that a functional network formed by the USH1B, C, D and G proteins is responsible for the correct cohesion of the hair bundle. PMID- 12588795 TI - A mouse model for hereditary hemorrhagic telangiectasia (HHT) type 2. AB - Hereditary hemorrhagic telangiectasia (HHT) is an autosomal-dominant disorder characterized by the age-dependent development of focal arteriovenous malformations and telangiectases. HHT type 2 is caused by loss of function mutations in activin receptor-like kinase 1 (ACVRL1 or ALK1). However, the factors that initiate lesion formation and those that influence disease progression remain unknown. Because heterozygous mice contain the appropriate genotype for an animal model of this disorder, mice heterozygous for a loss-of function mutation in Acvrl1 were carefully examined for an HHT-like phenotype. These mice developed age-dependent vascular lesions in the skin, extremities, oral cavity and in the internal organs (lung, liver, intestine, spleen and brain), similar to those seen in HHT patients. Major histopathological features of the lesions included thin-walled dilated vessels in close proximity to each other, hemorrhage and fibrosis. Similar to HHT patients, the mice also exhibited gastrointestinal bleeding, as evidenced by positive fecal occult blood tests. An Acvrl1(+/-) mouse with profound liver involvement also displayed a secondary cardiac phenotype, similar to that observed in human patients. The similarity of affected organs, age-dependent penetrance, histological similarity of the lesions and recapitulation of a secondary phenotype suggest that the Acvrl1(+/-) mice are an appropriate animal model for the identification of additional genetic and environmental factors that cause pathology in HHT type 2 patients. In addition, studies utilizing this animal model can yield valuable information on the role of ALK1 in maintenance of adult vascular architecture including arteriovenous identity. PMID- 12588796 TI - Defective integrin switch and matrix composition at alpha 7-deficient myotendinous junctions precede the onset of muscular dystrophy in mice. AB - Force transmission at the myotendinous junction requires a strong link between the muscle cytoskeleton and the extracellular matrix. At the adult junction, two splice variants of the laminin-binding integrins, alpha7Abeta1D and alpha7Bbeta1D, are highly enriched. The alpha7 subunits are critical for the integrity of the junctional sarcolemma because integrin alpha7-deficient mice develop muscular dystrophy, primarily affecting this site of the muscle. Here, we report that beta1D integrin coimmunoprecipitates and colocalizes with the alpha5 subunit at alpha7-deficient junctions, but does not associate with alpha3, alpha6 or alphav integrins. By immunogold labelling we show that the basement membranes of integrin alpha7-deficient muscles recruit abnormally high levels of fibronectin, the ligand of alpha5beta1D. Finally, we demonstrate that alpha5beta1D is down-regulated at the normal postnatal junction and is displaced by alpha7beta1D. These results suggest that the alpha7 subunit is implicated in the down-regulation of alpha5beta1D and in the removal of fibronectin from the maturing myotendinous junction, thus providing an alpha7beta1D-based link to laminin. We propose that the persistence of alpha5beta1D in alpha7-deficient mice is not compatible with normal muscle function and leads to muscle wasting. PMID- 12588797 TI - Specific progressive cAMP reduction implicates energy deficit in presymptomatic Huntington's disease knock-in mice. AB - Defects in gene transcription and mitochondrial function have been implicated in the dominant disease process that leads to the loss of striatal neurons in Huntington's disease (HD). Here we have used precise genetic HD mouse and striatal cell models to investigate the hypothesis that decreased cAMP responsive element (CRE)-mediated gene transcription may reflect impaired energy metabolism. We found that reduced CRE-signaling in Hdh(Q111) striatum, monitored by brain derived neurotrophic factor and phospho-CRE binding protein (CREB), predated inclusion formation. Furthermore, cAMP levels in Hdh(Q111) striatum declined from an early age (10 weeks), and cAMP was significantly decreased in HD postmortem brain and lymphoblastoid cells, attesting to a chronic deficit in man. Reduced CRE-signaling in cultured STHdh(Q111) striatal cells was associated with cytosolic CREB binding protein that mirrored diminished cAMP synthesis. Moreover, mutant cells exhibited mitochondrial respiratory chain impairment, evidenced by decreased ATP and ATP/ADP ratio, impaired MTT conversion and heightened sensitivity to 3-nitropropionic acid. Thus, our findings strongly suggest that impaired ATP synthesis and diminished cAMP levels amplify the early HD disease cascade by decreasing CRE-regulated gene transcription and altering energy dependent processes essential to neuronal cell survival. PMID- 12588798 TI - A major locus on mouse chromosome 18 controls XX sex reversal in Odd Sex (Ods) mice. AB - We have previously reported a dominant mouse mutant, Odd sex (Ods), in which XX Ods/+ mice on the FVB/N background show complete sex reversal, associated with expression of Sox9 in the fetal gonads. Remarkably, when crossed to the A/J strain approximately 95% of the (AXFVB) F(1) XX Ods/+ mice developed as fully fertile, phenotypic females, the remainder developing as males or hermaphrodites. Using a (AXFVB) F(2) population, we conducted a genome-wide linkage scan to identify the number and chromosomal location of potential Ods modifier genes. A single major locus termed Odsm1 was mapped to chromosome 18, tightly linked to D18Mit189 and D18Mit210. Segregation at this locus could account for the presence of sex reversal in 100% of XX Ods/+ mice which develop as males, for the absence of sex reversal in approximately 92% of XX Ods/+ mice which develop as females, and for the mixed sexual phenotype in approximately 72% of XX Ods/+ mice that develop with ambiguous genitalia. We propose that homozygosity for the FVB derived allele strongly favors Ods sex reversal, whereas homozygosity for the A/J derived allele inhibits it. In mice heterozygous at Odsm1, the phenotypic outcome, male, female or hermaphrodite, is determined by a complex interaction of several minor modifying loci. The close proximity of Smad2, Smad7 and Smad4 to D18Mit189/210 provides a potential mechanism through which Odsm1 might act. PMID- 12588799 TI - Parkin prevents mitochondrial swelling and cytochrome c release in mitochondria dependent cell death. AB - Parkin gene mutations have been implicated in autosomal-recessive early-onset parkinsonism and lead to specific degeneration of dopaminergic neurons in midbrain. To investigate the role of Parkin in neuronal cell death, we overproduced this protein in PC12 cells in an inducible manner. In this cell line, neuronally differentiated by nerve growth factor, Parkin overproduction protected against cell death mediated by ceramide, but not by a variety of other cell death inducers (H(2)O(2), 4-hydroxynonenal, rotenone, 6-OHDA, tunicamycin, 2 mercaptoethanol and staurosporine). Protection was abrogated by the proteasome inhibitor epoxomicin and disease-causing variants, indicating that it was mediated by the E3 ubiquitin ligase activity of Parkin. Interestingly, Parkin acted by delaying mitochondrial swelling and subsequent cytochrome c release and caspase-3 activation observed in ceramide-mediated cell death. Subcellular fractionation demonstrated enrichment of Parkin in the mitochondrial fraction and its association with the outer mitochondrial membrane. Together, these results suggest that Parkin may promote the degradation of substrates localized in mitochondria and involved in the late mitochondrial phase of ceramide-mediated cell death. Loss of this function may underlie the degeneration of nigral dopaminergic neurons in patients with Parkin mutations. PMID- 12588800 TI - Worldwide distribution and broader clinical spectrum of muscle-eye-brain disease. AB - Muscle-eye-brain disease (MEB), an autosomal recessive disorder prevalent in Finland, is characterized by congenital muscular dystrophy, brain malformation and ocular abnormalities. Since the MEB phenotype overlaps substantially with those of Fukuyama-type congenital muscular dystrophy (FCMD) and Walker-Warburg syndrome (WWS), these three diseases are thought to result from a similar pathomechanism. Recently, we showed that MEB is caused by mutations in the protein O-linked mannose beta1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) gene. We describe here the identification of seven novel disease-causing mutations in six of not only non-Finnish Caucasian but also Japanese and Korean patients with suspected MEB, severe FCMD or WWS. Including six previously reported mutations, the 13 disease-causing mutations we have found thus far are dispersed throughout the entire POMGnT1 gene. We also observed a slight correlation between the location of the mutation and clinical severity in the brain: patients with mutations near the 5' terminus of the POMGnT1 coding region show relatively severe brain symptoms such as hydrocephalus, while patients with mutations near the 3' terminus have milder phenotypes. Our results indicate that MEB may exist in population groups outside of Finland, with a worldwide distribution beyond our expectations, and that the clinical spectrum of MEB is broader than recognized previously. These findings emphasize the importance of considering MEB and searching for POMGnT1 mutations in WWS or other congenital muscular dystrophy patients worldwide. PMID- 12588801 TI - Loss of imprinting of IGF2 and H19 in osteosarcoma is accompanied by reciprocal methylation changes of a CTCF-binding site. AB - The adjacent insulin-like growth factor 2 (IGF2) and H19 genes are imprinted in most normal human tissues, but imprinting is often lost in tumors. The mechanisms involved in maintenance of imprinting (MOI) and loss of imprinting (LOI) are unresolved. We show here that osteosarcoma (OS) tumors with IGF2/H19 MOI exhibit allele-specific differential methylation of a CTCF-binding site upstream of H19. LOI of IGF2 or H19 in OS occurs in a mutually exclusive manner, and occurs with monoallelic expression of the other gene. Bisulfite sequencing reveals IGF2 LOI occurs with biallelic CpG methylation of the CTCF-binding site, while H19 LOI occurs with biallelic hypomethylation of this site. Our data demonstrate that IGF2 LOI and H19 LOI are accompanied by reciprocal methylation changes at a critical CTCF-binding site. We propose a model in which incomplete gain or loss of methylation at this CTCF-binding site during tumorigenesis explains the complex and often conflicting expression patterns of IGF2 and H19 in tumors. PMID- 12588802 TI - The voltage-gated potassium channel Kv1.3 regulates energy homeostasis and body weight. AB - Voltage-gated potassium (Kv) channels regulate cell membrane potential and control a variety of cellular processes. Kv1.3 channels are expressed in several tissues and believed to participate in cell volume regulation, apoptosis, T cell activation and renal solute homeostasis. Examination of Kv1.3-deficient mice (Kv1.3(-/-)), generated by gene targeting, revealed a previously unrecognized role for Kv1.3 in body weight regulation. Indeed, Kv1.3(-/-) mice weigh significantly less than control littermates. Moreover, knockout mice are protected from diet-induced obesity and gain significantly less weight than littermate controls when placed on a high-fat diet. While food intake did not differ significantly between Kv1.3(-/-) and controls, basal metabolic rate, measured at rest by indirect calorimetry, was significantly higher in knockout animals. These data indicate that Kv1.3 channels may participate in the pathways that regulate body weight and that channel inhibition increases basal metabolic rate. PMID- 12588803 TI - Mutations in the human melanocortin-4 receptor gene associated with severe familial obesity disrupts receptor function through multiple molecular mechanisms. AB - Mutations in the melanocortin-4 receptor gene (MC4R) represent the commonest monogenic cause of human obesity. However, information regarding the precise effects of such mutations on receptor function is very limited. We examined the functional properties of 12 different mutations in human MC4R that result in severe, familial, early-onset obesity. Of the nine missense mutants studied, four were completely unable to generate cAMP in response to ligand and five were partially impaired. Four showed evidence of impaired cell surface expression and six of reduced binding affinity for ligand. One mutation in the C-terminal tail, I316S, showed reduced affinity for alpha-MSH but retained normal affinity for the antagonist AgRP. None of the mutations inhibited signaling through co-transfected wild-type receptors. Thus, in the most comprehensive study to date of the functional properties of naturally occurring MC4R mutations we have (1) established that defective expression on the cell surface is a common mechanism impairing receptor function, (2) identified mutations which specifically affect ligand binding affinity thus aiding the definition of receptor structure-function relationships, (3) provided evidence against the notion that these receptor mutants act as dominant-negatives, and (4) identified a potentially novel molecular mechanism of receptor dysfunction whereby a mutation alters the relative affinities of a receptor for its natural agonist versus antagonist. PMID- 12588804 TI - Identification of CDH1 germline missense mutations associated with functional inactivation of the E-cadherin protein in young gastric cancer probands. AB - E-cadherin is involved in the formation of cell-junctions and the maintenance of epithelial integrity. Direct evidence of E-cadherin mutations triggering tumorigenesis has come from the finding of inactivating germline mutations of the gene (CDH1) in hereditary diffuse gastric cancer (HDGC). We screened a series of 66 young gastric cancer probands for germline CDH1 mutations, and two novel missense alterations together with an intronic variant were identified. We then analysed the functional significance of the two exonic missense variants found here as well as a third germline missense variant that we previously identified in a HGDC family. cDNAs encoding either the wild-type protein or mutant forms of E-cadherin were stably transfected into CHO (Chinese hamster ovary) E-cadherin negative cells. Transfected cell-lines were characterized in terms of aggregation, motility and invasion. We show that a proportion of apparently sporadic early-onset diffuse gastric carcinomas are associated with germline alterations of the E-cadherin gene. We also demonstrate that a proportion of missense variants are associated with significant functional consequences, suggesting that our cell model can be used as an adjunct in deciding on the potential pathogenic role of identified E-cadherin germline alterations. PMID- 12588807 TI - The pathophysiological consequences of somatostatin receptor internalization and resistance. AB - Somatostatin receptors expressed on tumor cells form the rationale for somatostatin analog treatment of patients with somatostatin receptor-positive neuroendocrine tumors. Nevertheless, although somatostatin analogs effectively control hormonal hypersecretion by GH-secreting pituitary adenomas, islet cell tumors, and carcinoid tumors, significant differences are observed among patients with respect to the efficacy of treatment. This may be related to a differential expression of somatostatin receptor subtypes among tumors. In addition, the property of somatostatin receptor subtypes to undergo agonist-induced internalization has important consequences for visualizing, as well as for therapy, of receptor-positive tumors using radioisotope- or chemotherapeutic compound-coupled somatostatin analogs. This review covers the pathophysiological role of somatostatin receptor subtypes in determining the efficacy of treatment of patients with somatostatin receptor-positive tumors using somatostatin analogs, as well as the preclinical and clinical consequences of agonist-induced receptor internalization for somatostatin receptor-targeted radio- and chemotherapy. Herein, the development and potential role of novel somatostatin analogs is discussed. PMID- 12588808 TI - The sodium/iodide Symporter (NIS): characterization, regulation, and medical significance. AB - The Na(+)/I(-) symporter (NIS) is an integral plasma membrane glycoprotein that mediates active I(-) transport into the thyroid follicular cells, the first step in thyroid hormone biosynthesis. NIS-mediated thyroidal I(-) transport from the bloodstream to the colloid is a vectorial process made possible by the selective targeting of NIS to the basolateral membrane. NIS also mediates active I(-) transport in other tissues, including salivary glands, gastric mucosa, and lactating mammary gland, in which it translocates I(-) into the milk for thyroid hormone biosynthesis by the nursing newborn. NIS provides the basis for the effective diagnostic and therapeutic management of thyroid cancer and its metastases with radioiodide. NIS research has proceeded at an astounding pace after the 1996 isolation of the rat NIS cDNA, comprising the elucidation of NIS secondary structure and topology, biogenesis and posttranslational modifications, transcriptional and posttranscriptional regulation, electrophysiological analysis, isolation of the human NIS cDNA, and determination of the human NIS genomic organization. Clinically related topics include the analysis of congenital I(-) transport defect-causing NIS mutations and the role of NIS in thyroid cancer. NIS has been transduced into various kinds of cancer cells to render them susceptible to destruction with radioiodide. Most dramatically, the discovery of endogenous NIS expression in more than 80% of human breast cancer samples has raised the possibility that radioiodide may be a valuable novel tool in breast cancer diagnosis and treatment. PMID- 12588810 TI - Peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1 alpha): transcriptional coactivator and metabolic regulator. AB - Investigations of biological programs that are controlled by gene transcription have mainly studied the regulation of transcription factors. However, there are examples in which the primary focus of biological regulation is at the level of a transcriptional coactivator. We have reviewed here the molecular mechanisms and biological programs controlled by the transcriptional coactivator peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1 alpha). Key cellular signals that control energy and nutrient homeostasis, such as cAMP and cytokine pathways, strongly activate PGC-1 alpha. Once PGC-1 alpha is activated, it powerfully induces and coordinates gene expression that stimulates mitochondrial oxidative metabolism in brown fat, fiber-type switching in skeletal muscle, and multiple aspects of the fasted response in liver. The regulation of these metabolic and cell fate decisions by PGC-1 alpha is achieved through specific interaction with a variety of transcription factors such as nuclear hormone receptors, nuclear respiratory factors, and muscle-specific transcription factors. PGC-1 alpha therefore constitutes one of the first and clearest examples in which biological programs are chiefly regulated by a transcriptional coactivator in response to environmental stimuli. Finally, PGC-1 alpha's control of energy homeostasis suggests that it could be a target for anti-obesity or diabetes drugs. PMID- 12588811 TI - When translation meets metabolism: multiple links to diabetes. AB - Type 2 diabetes is a polygenic disorder characterized by multiple biochemical defects including transcriptional, translational, and posttranslational abnormalities. Although major progress has been made in elucidation of factors at the transcriptional and posttranslational levels, defects at the translational level remain elusive. Mutation of a kinase that regulates translation initiation has been implicated in the etiology of a monogenic form of diabetes known as Wolcott-Rallison syndrome. Characterization of mice rendered deficient in eukaryotic initiation factors has provided model systems to study the involvement of translation in regulating insulin synthesis and secretion, hepatic function, peripheral insulin resistance, and diabetic complications. Recent progress in the understanding of endoplasmic reticulum overload by unfolded proteins has begun to uncover mechanisms leading to pancreatic beta-cell exhaustion. Future advances in this area may lead to identification of the missing links in the pathogenesis of beta-cell failures due to conditions such as hyperinsulinemia, hyperglycemia, and long-term treatment with sulfonylureas, and thus may identify novel therapeutic targets for diabetes. PMID- 12588812 TI - Management of simple nodular goiter: current status and future perspectives. AB - The simple nodular goiter, the etiology of which is multifactorial, encompasses the spectrum from the incidental asymptomatic small solitary nodule to the large intrathoracic goiter, causing pressure symptoms as well as cosmetic complaints. Its management is still the cause of considerable controversy. The mainstay in the diagnostic evaluation is related to functional and morphological characterization with serum TSH and (some kind of) imaging. Because malignancy is just as common in patients with a multinodular goiter as patients with a solitary nodule, we support the increasing use of fine-needle aspiration biopsy (cytology). Most patients need no treatment after malignancy is ruled out. In case of cosmetic or pressure symptoms, the choice in multinodular goiter stands between surgery, which is still the first choice, and radioiodine if uptake is adequate. In addition to surgery, the solitary nodule, whether hot or cold, can be treated with percutaneous ethanol injection therapy. If hot, radioiodine is the therapy of choice. Randomized studies are scarce, and the side effects of nonsurgical therapy are coming into focus. Therefore, the use of the optimum option in the individual patient cannot at present be based on evidence. However, we are of the view that levothyroxine, although widely used, should no longer be recommended routinely for this condition. Within a few years, the introduction of recombinant human TSH and laser therapy may profoundly alter the nonsurgical treatment of simple nodular goiter. PMID- 12588813 TI - Disseminating evidence-based mental health practices. PMID- 12588805 TI - The role of prolactin in mammary carcinoma. AB - The contribution of prolactin (PRL) to the pathogenesis and progression of human breast cancer at the cellular, transgenic, and epidemiological levels is increasingly appreciated. Acting at the endocrine and autocrine/paracrine levels, PRL functions to stimulate the growth and motility of human breast cancer cells. The actions of this ligand are mediated by at least six recognized PRL receptor isoforms found on, or secreted by, human breast epithelium. The PRL/PRL receptor complex associates with and activates several signaling networks that are shared with other members of the cytokine receptor superfamily. Coupled with the recently identified intranuclear function of PRL, these networks are integrated into the in vitro and in vivo actions induced by ligand. These findings indicate that antagonists of PRL/PRL receptor interaction or PRL receptor-associated signal transduction may be of considerable utility in the treatment of human breast cancer. PMID- 12588815 TI - 9% of a US community sample had a DSM-IV personality disorder. PMID- 12588814 TI - Strategies for implementing evidence-based practices in routine mental health settings. PMID- 12588816 TI - 22% of preschool children, 8% of primary school students and 3% of adolescents have tics. PMID- 12588817 TI - 17% of people with traumatic brain injury attempted suicide. PMID- 12588818 TI - Parents under-report children's suicide ideation and attempts. PMID- 12588820 TI - Existing psychological problems predict depression in primary care better than sociodemographic factors. PMID- 12588819 TI - Young men who do not drink, as well as those who drink heavily, have high levels of depression and distress. PMID- 12588821 TI - Mothers depression in early childhood increases the risk of adolescent anxiety and depression. PMID- 12588822 TI - There was no sex difference in genetic correlations between neuroticism and major depression. PMID- 12588823 TI - Persistent early childhood developmental impairment may be associated with later schizophreniform disorder. PMID- 12588824 TI - Childhood motor impairment is associated with male anxiety at 11 and 16 years. PMID- 12588825 TI - Markers of minor brain damage are associated with developmental performance in preschool children. PMID- 12588826 TI - Toddler temperament, cognition and caregiver interaction predict impulsive functioning. PMID- 12588827 TI - Three new childhood anxiety scales have comparable reliability and validity to traditional diagnostic scales. PMID- 12588828 TI - RBANS has reasonable test-retest reliability in schizophrenia. PMID- 12588829 TI - Review: in obsessive-compulsive disorder, clomipramine may be more effective than selective serotonin reuptake inhibitors after controlling for other factors. PMID- 12588830 TI - Long term citalopram prevents recurrent depression in the elderly and is well tolerated. PMID- 12588831 TI - Review: cognitive therapy may be no more effective than other bona fide psychological therapies for depression. PMID- 12588832 TI - Brief cognitive behavioural intervention delivered by nurses reduces overall symptoms in schizophrenia. PMID- 12588833 TI - 10 mg intramuscular olanzapine reduces acute agitation in schizophrenia more effectively than lower doses. PMID- 12588834 TI - Olanzapine reduces mania more effectively than divalproex, but has more adverse effects. PMID- 12588835 TI - Sertraline with or without interpersonal psychotherapy reduces dysthymia symptoms over psychotherapy alone. PMID- 12588837 TI - At 10 year follow up, brief interventions had no effect on harmful alcohol consumption. PMID- 12588836 TI - High dose buprenorphine may be an effective interim treatment for long term heroin users waiting for drug-assisted rehabilitation. PMID- 12588838 TI - Personality disorders have a differential effect on the outcome of anxiety disorders. PMID- 12588839 TI - The C. elegans che-1 gene encodes a zinc finger transcription factor required for specification of the ASE chemosensory neurons. AB - Chemotaxis to water-soluble chemicals such as NaCl is an important behavior of C. elegans when seeking food. ASE chemosensory neurons have a major role in this behavior. We show that che-1, defined by chemotaxis defects, encodes a zinc finger protein similar to the GLASS transcription factor required for photoreceptor cell differentiation in Drosophila, and that che-1 is essential for specification and function of ASE neurons. Expression of a che-1::gfp fusion construct was predominant in ASE. In che-1 mutants, expression of genes characterizing ASE such as seven-transmembrane receptors, guanylate cyclases and a cyclic-nucleotide gated channel is lost. Ectopic expression of che-1 cDNA induced expression of ASE-specific marker genes, a dye-filling defect in neurons other than ASE and dauer formation. PMID- 12588840 TI - Tbx4-Fgf10 system controls lung bud formation during chicken embryonic development. AB - The respiratory primordium is positioned and its territory is defined in the foregut. The visceral mesoderm of the respiratory primordium acquires the inducing potential that is necessary for endodermal budding morphogenesis and respiratory endoderm formation. Tbx4, a member of the T-box transcription factor gene family, was specifically expressed in the visceral mesoderm of the lung primordium. To analyze the function of Tbx4, we ectopically expressed Tbx4 in the visceral mesoderm of the foregut using in ovo electroporation. Ectopic Tbx4 induced ectopic bud formation in the esophagus by activating the expression of Fgf10. Conversely, interference of Tbx4 function resulted in repression of Fgf10 expression and in failure of lung bud formation. In addition, ectopic Tbx4 or Fgf10 also induced ectopic expression of Nkx2.1, a marker gene specific for the respiratory endoderm, in the underlying esophagus endoderm. When the border of the Tbx4 expression domain, which demarcates the respiratory tract and the esophagus, was disturbed by misexpression of Tbx4, formation of the tracheo esophageal septum failed. These results suggested that Tbx4 governs multiple processes during respiratory tract development; i.e. the initial endodermal bud formation, respiratory endoderm formation, and septation of the respiratory tract and the esophagus. PMID- 12588841 TI - The p27cip/kip ortholog dacapo maintains the Drosophila oocyte in prophase of meiosis I. AB - Animal oocytes undergo a highly conserved developmental arrest in prophase of meiosis I. Often this marks a period of rapid growth for the oocyte and is necessary to coordinate meiotic progression with the developmental events of oogenesis. In Drosophila, the oocyte develops within a 16-cell germline cyst. Throughout much of oogenesis, the oocyte remains in prophase of meiosis I. By contrast, its 15 mitotic sisters enter the endocycle and become polyploid in preparation for their role as nurse cells. How germline cysts establish and maintain these two independent cell cycles is unknown. We demonstrate a role for the p21(CIP)/p27(Kip1)/p57(Kip2)-like cyclin-dependent kinase inhibitor (cki) dacapo in the maintenance of the meiotic cycle in Drosophila oocytes. Our data indicate that it is through the differential regulation of the cki Dacapo that two modes of cell-cycle regulation are independently maintained within the common cytoplasm of ovarian cysts. PMID- 12588842 TI - Genome-wide identification of in vivo Drosophila Engrailed-binding DNA fragments and related target genes. AB - Chromatin immunoprecipitation after UV crosslinking of DNA/protein interactions was used to construct a library enriched in genomic sequences that bind to the Engrailed transcription factor in Drosophila embryos. Sequencing of the clones led to the identification of 203 Engrailed-binding fragments localized in intergenic or intronic regions. Genes lying near these fragments, which are considered as potential Engrailed target genes, are involved in different developmental pathways, such as anteroposterior patterning, muscle development, tracheal pathfinding or axon guidance. We validated this approach by in vitro and in vivo tests performed on a subset of Engrailed potential targets involved in these various pathways. Finally, we present strong evidence showing that an immunoprecipitated genomic DNA fragment corresponds to a promoter region involved in the direct regulation of frizzled2 expression by engrailed in vivo. PMID- 12588843 TI - Polarization of the C. elegans zygote proceeds via distinct establishment and maintenance phases. AB - Polarization of the C. elegans zygote along the anterior-posterior axis depends on cortically enriched (PAR) and cytoplasmic (MEX-5/6) proteins, which function together to localize determinants (e.g. PIE-1) in response to a polarizing cue associated with the sperm asters. Using time-lapse microscopy and GFP fusions, we have analyzed the localization dynamics of PAR-2, PAR-6, MEX-5, MEX-6 and PIE-1 in wild-type and mutant embryos. These studies reveal that polarization involves two genetically and temporally distinct phases. During the first phase (establishment), the sperm asters at one end of the embryo exclude the PAR-3/PAR 6/PKC3 complex from the nearby cortex, allowing the ring finger protein PAR-2 to accumulate in an expanding 'posterior' domain. Onset of the establishment phase involves the non-muscle myosin NMY-2 and the 14-3-3 protein PAR-5. The kinase PAR 1 and the CCCH finger proteins MEX-5 and MEX-6 also function during the establishment phase in a feedback loop to regulate growth of the posterior domain. The second phase begins after pronuclear meeting, when the sperm asters begin to invade the anterior. During this phase (maintenance), PAR-2 maintains anterior-posterior polarity by excluding the PAR-3/PAR-6/PKC3 complex from the posterior. These findings provide a model for how PAR and MEX proteins convert a transient asymmetry into a stably polarized axis. PMID- 12588844 TI - BDNF regulates spontaneous correlated activity at early developmental stages by increasing synaptogenesis and expression of the K+/Cl- co-transporter KCC2. AB - Spontaneous neural activity is a basic property of the developing brain, which regulates key developmental processes, including migration, neural differentiation and formation and refinement of connections. The mechanisms regulating spontaneous activity are not known. By using transgenic embryos that overexpress BDNF under the control of the nestin promoter, we show here that BDNF controls the emergence and robustness of spontaneous activity in embryonic hippocampal slices. Further, BDNF dramatically increases spontaneous co-active network activity, which is believed to synchronize gene expression and synaptogenesis in vast numbers of neurons. In fact, BDNF raises the spontaneous activity of E18 hippocampal neurons to levels that are typical of postnatal slices. We also show that BDNF overexpression increases the number of synapses at much earlier stages (E18) than those reported previously. Most of these synapses were GABAergic, and GABAergic interneurons showed hypertrophy and a 3-fold increase in GAD expression. Interestingly, whereas BDNF does not alter the expression of GABA and glutamate ionotropic receptors, it does raise the expression of the recently cloned K(+)/Cl(-) KCC2 co-transporter, which is responsible for the conversion of GABA responses from depolarizing to inhibitory, through the control of the Cl(-) potential. Together, results indicate that both the presynaptic and postsynaptic machineries of GABAergic circuits may be essential targets of BDNF actions to control spontaneous activity. The data indicate that BDNF is a potent regulator of spontaneous activity and co-active networks, which is a new level of regulation of neurotrophins. Given that BDNF itself is regulated by neuronal activity, we suggest that BDNF acts as a homeostatic factor controlling the emergence, complexity and networking properties of spontaneous networks. PMID- 12588845 TI - XOtx5b and XOtx2 regulate photoreceptor and bipolar fates in the Xenopus retina. AB - Photoreceptor and bipolar cells are molecularly related cell types in the vertebrate retina. XOtx5b is expressed in both photoreceptors and bipolars, while a closely related member of the same family of transcription factors, XOtx2, is expressed in bipolar cells only. Lipofection of retinal precursors with XOtx5b biases them toward photoreceptor fates whereas a similar experiment with XOtx2 promotes bipolar cell fates. Domain swap experiments show that the ability to specify different cell fates is largely contained in the divergent sequence C terminal to the homeodomain, while the more homologous N-terminal and homeodomain regions of both genes, when fused to VP16 activators, promote only photoreceptor fates. XOtx5b is closely related to Crx and like Crx it drives expression from an opsin reporter in vivo. XOtx2 suppresses this XOtx5b-driven reporter activity providing a possible explanation for why bipolars do not express opsin. Similarly, co-lipofection of XOtx2 with XOtx5b overrides the latter's ability to promote photoreceptor fates and the combination drives bipolar fates. The results suggest that the shared and divergent parts of these homologous genes may be involved in specifying the shared and distinct characters of related cell types in the vertebrate retina. PMID- 12588846 TI - Dp1 is required for extra-embryonic development. AB - Release of E2F1/DP1 heterodimers from repression mediated by the retinoblastoma tumor suppressor (pRB) triggers cell cycle entry into S phase, suggesting that E2F1 and DP1 proteins must act in unison, either to facilitate or to suppress cell-cycle progression. In stark contrast to the milder phenotypes that result from inactivation of E2Fs, we report that loss of Dp1 leads to death in utero because of the failure of extra-embryonic development. Loss of Dp1 compromises the trophectoderm-derived tissues - specifically, the expansion of the ectoplacental cone and chorion, and endoreduplication in trophoblast giant cells. Inactivation of p53 is unable to rescue the Dp1-deficient embryonic lethality. Thus, DP1 is absolutely required for extra-embryonic development and consequently embryonic survival, consistent with E2F/DP1 normally acting to promote growth in vivo. PMID- 12588847 TI - Dock and Pak regulate olfactory axon pathfinding in Drosophila. AB - The convergence of olfactory axons expressing particular odorant receptor (Or) genes on spatially invariant glomeruli in the brain is one of the most dramatic examples of precise axon targeting in developmental neurobiology. The cellular and molecular mechanisms by which olfactory axons pathfind to their targets are poorly understood. We report here that the SH2/SH3 adapter Dock and the serine/threonine kinase Pak are necessary for the precise guidance of olfactory axons. Using antibody localization, mosaic analyses and cell-type specific rescue, we observed that Dock and Pak are expressed in olfactory axons and function autonomously in olfactory neurons to regulate the precise wiring of the olfactory map. Detailed analyses of the mutant phenotypes in whole mutants and in small multicellular clones indicate that Dock and Pak do not control olfactory neuron (ON) differentiation, but specifically regulate multiple aspects of axon trajectories to guide them to their cognate glomeruli. Structure/function studies show that Dock and Pak form a signaling pathway that mediates the response of olfactory axons to guidance cues in the developing antennal lobe (AL). Our findings therefore identify a central signaling module that is used by ONs to project to their cognate glomeruli. PMID- 12588848 TI - Bicarbonate actions on flagellar and Ca2+ -channel responses: initial events in sperm activation. AB - At mating, mammalian sperm are diluted in the male and female reproductive fluids, which brings contact with HCO(3)(-) and initiates several cellular responses. We have identified and studied two of the most rapid of these responses. Stop-motion imaging and flagellar waveform analysis show that for mouse epididymal sperm in vitro, the resting flagellar beat frequency is 2-3 Hz at 22-25 degrees C. Local perfusion with HCO(3)(-) produces a robust, reversible acceleration to 7 Hz or more. At 15 mM the action of HCO(3)(-) begins within 5 seconds and is near-maximal by 30 seconds. The half-times of response are 8.8+/ 0.2 seconds at 15 mM HCO(3)(-) and 17.5+/-0.4 seconds at 1 mM HCO(3)(-). Removal of external HCO(3)(-) allows a slow return to basal beat frequency over approximately 10 minutes. Increases in beat symmetry accompany the accelerating action of HCO(3)(-). As in our past work, HCO(3)(-) also facilitates opening of voltagegated Ca(2+) channels, increasing the depolarization-evoked rate of rise of intracellular Ca(2+) concentration by more than fivefold. This action also is detectable at 1 mM HCO(3)(-) and occurs with an apparent halftime of approximately 60 seconds at 15 mM HCO(3)(-). The dual actions of HCO(3)(-) respond similarly to pharmacological intervention. Thus, the phosphodiesterase inhibitor IBMX promotes the actions of HCO(3)(-) on flagellar and channel function, and the protein kinase A inhibitor H89 blocks these actions. In addition, a 30 minute incubation with 60 micro M cAMP acetoxylmethyl ester increases flagellar beat frequency to nearly 7 Hz and increases the evoked rates of rise of intracellular Ca(2+) concentration from 17+/-4 to 41+/-6 nM second( 1). However, treatment with several other analogs of cAMP produces only scant evidence of the expected mimicry or blockade of the actions of HCO(3)(-), perhaps as a consequence of limited permeation. Our findings indicate a requirement for cAMP-mediated protein phosphorylation in the enhancement of flagellar and channel functions that HCO(3)(-) produces during sperm activation. PMID- 12588849 TI - A role for the mesenchymal T-box gene Brachyury in AER formation during limb development. AB - During limb development, several signaling centers organize limb pattern. One of these, the apical ectodermal ridge (AER), is critical for proximodistal limb outgrowth mediated by FGFs. Signals from the underlying mesoderm, including WNTs and FGFs, regulate early steps of AER induction. Ectodermal factors, particularly En1, play a critical role in regulating morphogenesis of a mature, compact AER along the distal limb apex, from a broad ventral ectodermal precursor domain. Contribution of mesodermal factors to the morphogenesis of a mature AER is less clear. We previously noted that the chick T gene (Brachyury), the prototypical T box transcription factor, is expressed in the limb bud as well as axial mesoderm and primitive streak. Here we show that T is expressed in lateral plate mesoderm at the onset of limb bud formation and subsequently in the subridge mesoderm beneath the AER. Retroviral misexpression of T in chick results in anterior extension of the AER and subsequent limb phenotypes consistent with augmented AER extent and function. Analysis of markers for functional AER in mouse T(-/-) null mutant limb buds reveals disrupted AER morphogenesis. Our data also suggest that FGF and WNT signals may operate both upstream and downstream of T. Taken together, the results show that T plays a role in the regulation of AER formation, particularly maturation, and suggest that T may also be a component of the epithelialmesenchymal regulatory loop involved in maintenance of a mature functioning AER. PMID- 12588850 TI - Endothelin 1-mediated regulation of pharyngeal bone development in zebrafish. AB - Endothelin 1 (Edn1), a secreted peptide expressed ventrally in the primordia of the zebrafish pharyngeal arches, is required for correct patterning of pharyngeal cartilage development. We have studied mutants and morpholino-injected larvae to examine the role of the Edn1 signal in patterning anterior pharyngeal arch bone development during the first week after fertilization. We observe a remarkable variety of phenotypic changes in dermal bones of the anterior arches after Edn1 reduction, including loss, size reduction and expansion, fusion and shape change. Notably, the changes that occur appear to relate to the level of residual Edn1. Mandibular arch dermal bone fusions occur with severe Edn1 loss. In the dorsal hyoid arch, the dermal opercle bone is usually absent when Edn1 is severely reduced and is usually enlarged when Edn1 is only mildly reduced, suggesting that the same signal can act both positively and negatively in controlling development of a single bone. Position also appears to influence the changes: a branchiostegal ray, a dermal hyoid bone normally ventral to the opercle, can be missing in the same arch where the opercle is enlarged. We propose that Edn1 acts as a morphogen; different levels pattern specific positions, shapes and sizes of bones along the dorso-ventral axis. Changes involving Edn1 may have occurred during actinopterygian evolution to produce the efficient gill-pumping opercular apparatus of teleosts. PMID- 12588851 TI - Two endothelin 1 effectors, hand2 and bapx1, pattern ventral pharyngeal cartilage and the jaw joint. AB - A conserved endothelin 1 signaling pathway patterns the jaw and other pharyngeal skeletal elements in mice, chicks and zebrafish. In zebrafish, endothelin 1 (edn1 or sucker) is required for formation of ventral cartilages and joints in the anterior pharyngeal arches of young larvae. Here we present genetic analyses in the zebrafish of two edn1 downstream targets, the bHLH transcription factor Hand2 and the homeobox transcription factor Bapx1, that mediate dorsoventral (DV) patterning in the anterior pharyngeal arches. First we show that edn1-expressing cells in the first (mandibular) and second (hyoid) pharyngeal arch primordia are located most ventrally and surrounded by hand2-expressing cells. Next we show that along the DV axis of the early first arch primordia, bapx1 is expressed in an intermediate domain, which later marks the jaw joint, and this expression requires edn1 function. bapx1 function is required for formation of the jaw joint, the joint-associated retroarticular process of Meckel's cartilage, and the retroarticular bone. Jaw joint expression of chd and gdf5 also requires bapx1 function. Similar to edn1, hand2 is required for ventral pharyngeal cartilage formation. However, the early ventral arch edn1-dependent expression of five genes (dlx3, EphA3, gsc, msxe and msxb) are all present in hand2 mutants. Further, msxe and msxb are upregulated in hand2 mutant ventral arches. Slightly later, an edn1-dependent ventral first arch expression domain of gsc is absent in hand2 mutants, providing a common downstream target of edn1 and hand2. In hand2 mutants, bapx1 expression is present at the joint region, and expanded ventrally. In addition, expression of eng2, normally restricted to first arch dorsal mesoderm, expands ventrally in hand2 and edn1 mutants. Thus, ventral pharyngeal specification involves repression of dorsal and intermediate (joint region) fates. Together our results reveal two critical edn1 effectors that pattern the vertebrate jaw: hand2 specifies ventral pharyngeal cartilage of the lower jaw and bapx1 specifies the jaw joint. PMID- 12588852 TI - The zinc-finger protein CNBP is required for forebrain formation in the mouse. AB - Mouse mutants have allowed us to gain significant insight into axis development. However, much remains to be learned about the cellular and molecular basis of early forebrain patterning. We describe a lethal mutation mouse strain generated using promoter-trap mutagenesis. The mutants exhibit severe forebrain truncation in homozygous mouse embryos and various craniofacial defects in heterozygotes. We show that the defects are caused by disruption of the gene encoding cellular nucleic acid binding protein (CNBP); Cnbp transgenic mice were able to rescue fully the mutant phenotype. Cnbp is first expressed in the anterior visceral endoderm (AVE) and, subsequently, in the anterior definitive endoderm (ADE), anterior neuroectoderm (ANE), anterior mesendoderm (AME), headfolds and forebrain. In Cnbp(-/-) embryos, the visceral endoderm remains in the distal tip of the conceptus and the ADE fails to form, whereas the node and notochord form normally. A substantial reduction in cell proliferation was observed in the anterior regions of Cnbp(-/-) embryos at gastrulation and neural-fold stages. In these regions, Myc expression was absent, indicating CNBP targets Myc in rostral head formation. Our findings demonstrate that Cnbp is essential for the forebrain induction and specification. PMID- 12588853 TI - Cell fate specification and competence by Coco, a maternal BMP, TGFbeta and Wnt inhibitor. AB - Patterning of the pre-gastrula embryo and subsequent neural induction post gastrulation are very complex and intricate processes of which little, until recently, has been understood. The earliest decision in neural development, the choice between epidermal or neural fates, is regulated by bone morphogenetic protein (BMP) signaling within the ectoderm. Inhibition of BMP signaling is sufficient for neural induction. Many secreted BMP inhibitors are expressed exclusively within the organizer of the Xenopus gastrula embryo and therefore are predicted to act as bona fide endogenous neural inducers. Other cell-autonomous inhibitors of the BMP pathway are more widely expressed, such as the inhibitory Smads, Smad6 and Smad7. In this report we describe the biological and biochemical characterization of 51-B6, a novel member of Cerberus/Dan family of secreted BMP inhibitors, which we identified in a screen for Smad7-induced genes. This gene is expressed maternally in an animal to vegetal gradient, and its expression levels decline rapidly following gastrulation. In contrast to known BMP inhibitors, 51 B6 is broadly expressed in the ectoderm until the end of gastrulation. The timing, pattern of expression, and activities of this gene makes it unique when compared to other BMP/TGFbeta/Wnt secreted inhibitors which are expressed only zygotically and maintained post-gastrulation. We propose that a function of 51-B6 is to block BMP and TGFbeta signals in the ectoderm in order to regulate cell fate specification and competence prior to the onset of neural induction. In addition, we demonstrate that 51-B6 can act as a neural inducer and induce ectopic head-like structures in neurula staged embryos. Because of this embryological activity, we have renamed this clone Coco, after the Spanish word meaning head. PMID- 12588854 TI - Delta-Notch signaling controls the generation of neurons/glia from neural stem cells in a stepwise process. AB - We examined the role of Notch signaling on the generation of neurons and glia from neural stem cells by using neurospheres that are clonally derived from neural stem cells. Neurospheres prepared from Dll1(lacZ/lacZ) mutant embryos segregate more neurons at the expense of both oligodendrocytes and astrocytes. This mutant phenotype could be rescued when Dll1(lacZ/lacZ) spheres were grown and/or differentiated in the presence of conditioned medium from wild-type neurospheres. Temporal modulation of Notch by soluble forms of ligands indicates that Notch signaling acts in two steps. Initially, it inhibits the neuronal fate while promoting the glial cell fate. In a second step, Notch promotes the differentiation of astrocytes, while inhibiting the differentiation of both neurons and oligodendrocytes. PMID- 12588855 TI - Hedgehog signalling is required for correct anteroposterior patterning of the zebrafish otic vesicle. AB - Currently, few factors have been identified that provide the inductive signals necessary to transform the simple otic placode into the complex asymmetric structure of the adult vertebrate inner ear. We provide evidence that Hedgehog signalling from ventral midline structures acts directly on the zebrafish otic vesicle to induce posterior otic identity. We demonstrate that two strong Hedgehog pathway mutants, chameleon (con(tf18b)) and slow muscle omitted (smu(b641)) exhibit a striking partial mirror image duplication of anterior otic structures, concomitant with a loss of posterior otic domains. These effects can be phenocopied by overexpression of patched1 mRNA to reduce Hedgehog signalling. Ectopic activation of the Hedgehog pathway, by injection of sonic hedgehog or dominant-negative protein kinase A RNA, has the reverse effect: ears lose anterior otic structures and show a mirror image duplication of posterior regions. By using double mutants and antisense morpholino analysis, we also show that both Sonic hedgehog and Tiggy-winkle hedgehog are involved in anteroposterior patterning of the zebrafish otic vesicle. PMID- 12588856 TI - Rac promotes epithelial cell rearrangement during tracheal tubulogenesis in Drosophila. AB - Cell rearrangement, accompanied by the rapid assembly and disassembly of cadherin mediated cell adhesions, plays essential roles in epithelial morphogenesis. Various in vitro and cell culture studies on the small GTPase Rac have suggested it to be a key regulator of cell adhesion, but this notion needs to be verified in the context of embryonic development. We used the tracheal system of Drosophila to investigate the function of Rac in the epithelial cell rearrangement, with a special attention to its role in regulating epithelial cadherin activity. We found that a reduced Rac activity led to an expansion of cell junctions in the embryonic epidermis and tracheal epithelia, which was accompanied by an increase in the amount of Drosophila E-Cadherin-Catenin complexes by a post-transcriptional mechanism. Reduced Rac activity inhibited dynamic epithelial cell rearrangement. Hyperactivation of Rac, on the other hand, inhibited assembly of newly synthesized E-Cadherin into cell junctions and caused loss of tracheal cell adhesion, resulting in cell detachment from the epithelia. Thus, in the context of Drosophila tracheal development, Rac activity must be maintained at a level necessary to balance the assembly and disassembly of E Cadherin at cell junctions. Together with its role in cell motility, Rac regulates plasticity of cell adhesion and thus ensures smooth remodeling of epithelial sheets into tubules. PMID- 12588857 TI - Quiescent center formation in maize roots is associated with an auxin-regulated oxidizing environment. AB - Embedded within the meristem of all Angiosperm roots is a population of slowly dividing cells designated the quiescent center (QC). In maize roots the QC can constitute upwards of 800-1200 cells, most of which spend an extended period of time (180-200 hours) in the G(1) phase of the cell cycle. How the QC forms and is maintained is not known. Here we report that cells of the QC are characterized by their highly oxidized status. Glutathione and ascorbic acid occur predominately in the oxidized forms in the QC. This is contrasted with the status of these redox intermediates in adjacent, rapidly dividing cells in the root meristem, in which the reduced forms of these two species are favored. Using a redox sensitive fluorescent dye we were able to visualize an overall oxidizing environment in the QC, and we also made comparisons with the adjacent, rapidly dividing cells in the root meristem. Altering the distribution of auxin and the location of the auxin maximum in the root tip activates the QC, and cells leave G(1) and enter mitosis. Commencement of relatively more rapid cell division in the QC is preceded by changes in the overall redox status of the QC, which becomes less oxidizing. We discuss how the position of the auxin maximum may influence the redox status of the QC and thereby modulate the cell cycle. PMID- 12588858 TI - Induction of neuron-specific glycosylation by Tollo/Toll-8, a Drosophila Toll like receptor expressed in non-neural cells. AB - Specific glycan expression is an essential characteristic of developing tissues. Our molecular characterization of a mutation that abolishes neural-specific glycosylation in the Drosophila embryo demonstrates that cellular interactions influence glycan expression. The HRP epitope is an N-linked oligosaccharide expressed on a subset of neuronal glycoproteins. Embryos homozygous for the TM3 balancer chromosome lack neural HRP-epitope expression. Genetic and molecular mapping of the relevant locus reveals that Tollo/Toll-8, a member of the Toll like receptor family, is altered on the TM3 chromosome. In wild-type embryos, Tollo/Toll-8 is expressed by ectodermal cells that surround differentiating neurons and precedes HRP-epitope appearance. Re-introduction of Tollo/Toll-8 into null embryos rescues neural-specific glycan expression. Thus, loss of an ectodermal cell surface protein alters glycosylation in juxtaposed differentiating neurons. The portfolio of expressed oligosaccharides in a cell reflects its identity and also influences its interactions with other cells and with pathogens. Therefore, the ability to induce specific glycan expression complements the previously identified developmental and innate immune functions of Toll-like receptors. PMID- 12588859 TI - Developing with lethal RA levels: genetic ablation of Rarg can restore the viability of mice lacking Cyp26a1. AB - We have previously reported that the retinoic acid (RA) catabolizing enzyme CYP26A1 plays an important role in protecting tail bud tissues from inappropriate exposure to RA generated in the adjacent trunk tissues by RALDH2, and that Cyp26a1-null animals exhibit spina bifida and caudal agenesis. We now show that, in the absence of Cyp26a1, retinoic acid receptor gamma (RARgamma) mediates ectopic RA-signaling in the tail bud. We also show that activated RARgamma results in downregulation of Wnt3a and Fgf8, which integrate highly conserved signaling pathways known for their role in specifying caudal morphogenesis. Ablation of the gene for RARgamma (Rarg) rescues Cyp26a1-null mutant animals from caudal regression and embryonic lethality, thus demonstrating that CYP26A1 suppresses the RA-mediated downregulation of WNT3A and FGF8 signaling pathways by eliminating ectopic RA in gastrulating tail bud mesoderm. PMID- 12588860 TI - Ca2+ oscillations at fertilization in mammals are regulated by the formation of pronuclei. AB - In mammals, the sperm triggers a series of cytosolic Ca(2+) oscillations that continue for approximately 4 hours, stopping close to the time of pronucleus formation. Ca(2+) transients are also seen in fertilized embryos during the first mitotic division. The mechanism that controls this pattern of sperm-induced Ca(2+) signalling is not known. Previous studies suggest two possible mechanisms: first, regulation of Ca(2+) oscillations by M-phase kinases; and second, regulation by the presence or absence of an intact nucleus. We describe experiments in mouse oocytes that differentiate between these mechanisms. We find that Ca(2+) oscillations continue after Cdk1-cyclin B1 activity falls at the time of polar body extrusion and after MAP kinase has been inhibited with UO126. This suggests that M-phase kinases are not necessary for continued Ca(2+) oscillations. A role for pronucleus formation in regulating Ca(2+) signalling is demonstrated in experiments where pronucleus formation is inhibited by microinjection of a lectin, WGA, without affecting the normal inactivation of the M-phase kinases. In oocytes with no pronuclei but with low M-phase kinase activity, sperm-induced Ca(2+) oscillations persist for nearly 10 hours. Furthermore, a dominant negative importin beta that inhibits nuclear transport, also prevents pronucleus formation and causes Ca(2+) oscillations that continue for nearly 12 hours. During mitosis, fluorescent tracers that mark nuclear envelope breakdown and the subsequent reformation of nuclei in the newly formed two-cell embryo establish that Ca(2+) oscillations are generated only in the absence of a patent nuclear membrane. We conclude by suggesting a model where nuclear sequestration and release of a Ca(2+)-releasing activity contributes to the temporal organization of Ca(2+) transients in meiosis and mitosis in mice. PMID- 12588861 TI - Drosophila necrotic mutations mirror disease-associated variants of human serpins. AB - Polymerization of members of the serpin superfamily underlies diseases as diverse as cirrhosis, angioedema, thrombosis and dementia. The Drosophila serpin Necrotic controls the innate immune response and is homologous to human alpha(1) antitrypsin. We show that necrotic mutations that are identical to the Z deficiency variant of alpha(1)-antitrypsin form urea-stable polymers in vivo. These necrotic mutations are temperature sensitive, which is in keeping with the temperature-dependent polymerization of serpins in vitro and the role of childhood fevers in exacerbating liver disease in Z alpha-antitrypsin deficiency. In addition, we identify two nec mutations homologous to an antithrombin point mutation that is responsible for neonatal thrombosis. Transgenic flies carrying an S>F amino-acid substitution equivalent to that found in Siiyama-variant antitrypsin (nec(S>F.UAS)) fail to complement nec-null mutations and demonstrate a dominant temperature-dependent inactivation of the wild-type nec allele. Taken together, these data establish Drosophila as a powerful system to study serpin polymerization in vivo. PMID- 12588862 TI - The human L(3)MBT polycomb group protein is a transcriptional repressor and interacts physically and functionally with TEL (ETV6). AB - H-L(3)MBT, the human homolog of the Drosophila lethal(3)malignant brain tumor protein, is a member of the polycomb group (PcG) of proteins, which function as transcriptional regulators in large protein complexes. Homozygous mutations in the l(3)mbt gene cause brain tumors in Drosophila, identifying l(3)mbt as a tumor suppressor gene. The h-l(3)mbt gene maps to chromosome 20q12, within a common deleted region associated with myeloid hematopoietic malignancies. H-L(3)MBT contains three repeats of 100 residues called MBT repeats, whose function is unknown, and a C-terminal alpha-helical structure, the SPM (SCM, PH, MBT domain, which is structurally similar to the SAM (sterile alpha motif) protein-protein interaction domain, found in several ETS transcription factors, including TEL (translocation Ets leukemia). We report that H-L(3)MBT is a transcriptional repressor and that its activity is largely dependent on the presence of a region containing the three MBT repeats. H-L(3)MBT acts as a histone deacetylase independent transcriptional repressor, based on its lack of sensitivity to trichostatin A. We found that H-L(3)MBT binds in vivo to TEL, and we have mapped the region of interaction to their respective SPM/SAM domains. We show that the ability of TEL to repress TEL-responsive promoters is enhanced by the presence of H-L(3)MBT, an effect dependent on the H-L(3)MBT and the TEL interacting domains. These experiments suggest that histone deacetylase-independent transcriptional repression by TEL depends on the recruitment of PcG proteins. We speculate that the interaction of TEL with H-L(3)MBT can direct a PcG complex to genes repressed by TEL, stabilizing their repressed state. PMID- 12588863 TI - Selective regulation of ptsG expression by Fis. Formation of either activating or repressing nucleoprotein complex in response to glucose. AB - Transcription of ptsG encoding glucose-specific permease, enzyme IICB(Glc), in Escherichia coli is initiated from two promoters, P1 and P2. ptsG transcription is repressed by Mlc, a glucose-inducible regulator of carbohydrate metabolism. The regulation of ptsG P1 transcription is also under positive control by cyclic AMP receptor protein and cyclic AMP complex (CRP.cAMP) as observed in other Mlc regulon. We report here that Fis, one of the nucleoid-associated proteins, plays a key role in glucose induction of Mlc regulon. ptsG transcription was induced when wild-type cells were grown in the presence of glucose. However, in a fis mutant, the basal level of ptsG transcription was higher but decreased when cells were grown in the presence of glucose, which implies the possibility of regulatory interactions among Fis, Mlc, and CRP.cAMP. Footprinting experiments with various probes and transcription assays revealed that Fis assists both Mlc repression and CRP.cAMP activation of ptsG P1 through the formation of Fis.CRP.Mlc or Fis.CRP nucleoprotein complexes at ptsG P1 promoter depending on the availability of glucose in the growth medium. ptsG P2 transcription was inhibited by Fis and Mlc. Tighter Mlc repression and enhanced CRP.cAMP activation of ptsG P1 by Fis enable cells to regulate Mlc regulon efficiently by selectively controlling the concentration of enzyme IICB(Glc) that modulates Mlc activity. PMID- 12588864 TI - Characterization of the heparin binding sites in human apolipoprotein E. AB - Apolipoprotein (apo) E mediates lipoprotein remnant clearance via interaction with cell-surface heparan sulfate proteoglycans. Both the 22-kDa N-terminal domain and 10-kDa C-terminal domain of apoE contain a heparin binding site; the N terminal site overlaps with the low density lipoprotein receptor binding region and the C-terminal site is undefined. To understand the molecular details of the apoE-heparin interaction, we defined the microenvironments of all 12 lysine residues in intact apoE3 and examined their relative contributions to heparin binding. Nuclear magnetic resonance measurements showed that, in apoE3 dimyristoyl phosphatidylcholine discs, Lys-143 and -146 in the N-terminal domain and Lys-233 in the C-terminal domain have unusually low pK(a) values, indicating high positive electrostatic potential around these residues. Binding experiments using heparin-Sepharose gel demonstrated that the lipid-free 10-kDa fragment interacted strongly with heparin and a point mutation K233Q largely abolished the binding, indicating that Lys-233 is involved in heparin binding and that an unusually basic lysine microenvironment is critical for the interaction with heparin. With lipidated apoE3, it is confirmed that the Lys-233 site is completely masked and the N-terminal site mediates heparin binding. In addition, mutations of the two heparin binding sites in intact apoE3 demonstrated the dominant role of the N-terminal site in the heparin binding of apoE even in the lipid-free state. These results suggest that apoE interacts predominately with cell-surface heparan sulfate proteoglycans through the N-terminal binding site. However, Lys-233 may be involved in the binding of apoE to certain cell-surface sites, such as the protein core of biglycan. PMID- 12588865 TI - Inhibition of transfer messenger RNA aminoacylation and trans-translation by aminoglycoside antibiotics. AB - Transfer messenger RNA (tmRNA) directs the modification of proteins of which the biosynthesis has stalled or has been interrupted. Here, we report that aminoglycosides can interfere with this quality control system in bacteria, termed trans-translation. Neomycin B is the strongest inhibitor of tmRNA aminoacylation with alanine (K(i) value of approximately 35 micro m), an essential step during trans-translation. The binding sites of neomycin B do not overlap with the identity determinants for alanylation, but the aminoglycoside perturbs the conformation of the acceptor stem that contains the aminoacylation signals. Aminoglycosides reduce the conformational freedom of the transfer RNA like domain of tmRNA. Additional contacts between aminoglycosides and tmRNA are within the tag reading frame, probably also disturbing reprogramming of the stalled ribosomes prior protein tagging. Aminoglycosides impair tmRNA aminoacylation in the presence of all of the transfer RNAs from Escherichia coli, small protein B, and elongation factor Tu, but when both proteins are present, the inhibition constant is 1 order of magnitude higher. SmpB and elongation factor Tu have RNA chaperone activities, ensuring that tmRNA adopts an optimal conformation during aminoacylation. PMID- 12588866 TI - Non-steroidal anti-inflammatory drugs inhibit nitric oxide-induced apoptosis and dedifferentiation of articular chondrocytes independent of cyclooxygenase activity. AB - Nitric oxide (NO) causes apoptosis and dedifferentiation of articular chondrocytes by the modulation of extracellular signal-regulated kinase (ERK), p38 kinase, and protein kinase C (PKC) alpha and -zeta. In this study, we investigated the effects and mechanisms of non-steroidal anti-inflammatory drugs (NSAIDs), such as indomethacin, ketoprofen, ibuprofen, sulindac sulfide, and flurbiprofen, in NO-induced apoptosis and dedifferentiation of articular chondrocytes. We found that all of the examined NSAIDs inhibited apoptosis and dedifferentiation. NO production in chondrocytes caused activation of ERK-1/2 and p38 kinase, which oppositely regulate apoptosis and dedifferentiation. NO production also caused inhibition of PKCalpha and -zeta independent of and dependent on, respectively, p38 kinase, which is required for apoptosis and dedifferentiation. Among the signaling molecules modulated by NO, NSAIDs blocked NO-induced activation of p38 kinase, potentiated ERK activation, and blocked inhibition of PKCalpha and -zeta. NSAIDs also inhibited some of the apoptotic signaling that is downstream of p38 kinase and PKC, such as NFkappaB activation, p53 accumulation, and caspase-3 activation. The inhibitory effects of NSAIDs on apoptosis and dedifferentiation were independent of the inhibition of cyclooxygenase (COX)-2 and prostaglandin E(2) (PGE(2)) production, as evidenced by the observation that specific inhibition of COX-2 activity and PGE(2) production or exogenous PGE(2) did not affect NO-induced apoptosis and dedifferentiation. Taken together, our results indicate that NSAIDs block NO induced apoptosis and dedifferentiation of articular chondrocytes by the modulation of ERK, p38 kinase, and PKCalpha and -zeta in a manner independent of their ability to inhibit COX-2 and PGE(2) production. PMID- 12588867 TI - Crystal structure of Thermotoga maritima alpha-glucosidase AglA defines a new clan of NAD+-dependent glycosidases. AB - Glycoside hydrolase family 4 represents an unusual group of glucosidases with a requirement for NAD+, divalent metal cations, and reducing conditions. The family is also unique in its inclusion of both alpha- and beta-specific enzymes. The alpha-glucosidase A, AglA, from Thermotoga maritima is a typical glycoside hydrolase family 4 enzyme, requiring NAD+ and Mn2+ as well as strongly reducing conditions for activity. Here we present the crystal structure of the protein complexed with NAD+ and maltose, refined at a resolution of 1.9 A. The NAD+ is bound to a typical Rossman fold NAD+-binding site, and the nicotinamide moiety is localized close to the maltose substrate. Within the active site the conserved Cys-174 and surrounding histidines are positioned to play a role in the hydrolysis reaction. The electron density maps indicate that Cys-174 is oxidized to a sulfinic acid. Most likely, the strongly reducing conditions are necessary to reduce the oxidized cysteine side chain. Notably, the canonical set of catalytic acidic residues common to other glucosidases is not present in the active site. This, combined with a high structural homology to NAD-dependent dehydrogenases, suggests an unusual and possibly unique mechanism of action for a glycoside-hydrolyzing enzyme. PMID- 12588868 TI - Ataxia-telangiectasia-mutated (ATM) and NBS1-dependent phosphorylation of Chk1 on Ser-317 in response to ionizing radiation. AB - In mammals, the ATM (ataxia-telangiectasia-mutated) and ATR (ATM and Rad3 related) protein kinases function as critical regulators of the cellular DNA damage response. The checkpoint functions of ATR and ATM are mediated, in part, by a pair of checkpoint effector kinases termed Chk1 and Chk2. In mammalian cells, evidence has been presented that Chk1 is devoted to the ATR signaling pathway and is modified by ATR in response to replication inhibition and UV induced damage, whereas Chk2 functions primarily through ATM in response to ionizing radiation (IR), suggesting that Chk2 and Chk1 might have evolved to channel the DNA damage signal from ATM and ATR, respectively. We demonstrate here that the ATR-Chk1 and ATM-Chk2 pathways are not parallel branches of the DNA damage response pathway but instead show a high degree of cross-talk and connectivity. ATM does in fact signal to Chk1 in response to IR. Phosphorylation of Chk1 on Ser-317 in response to IR is ATM-dependent. We also show that functional NBS1 is required for phosphorylation of Chk1, indicating that NBS1 might facilitate the access of Chk1 to ATM at the sites of DNA damage. Abrogation of Chk1 expression by RNA interference resulted in defects in IR-induced S and G(2)/M phase checkpoints; however, the overexpression of phosphorylation site mutant (S317A, S345A or S317A/S345A double mutant) Chk1 failed to interfere with these checkpoints. Surprisingly, the kinase-dead Chk1 (D130A) also failed to abrogate the S and G(2) checkpoint through any obvious dominant negative effect toward endogenous Chk1. Therefore, further studies will be required to assess the contribution made by phosphorylation events to Chk1 regulation. Overall, the data presented in the study challenge the model in which Chk1 only functions downstream from ATR and indicate that ATM does signal to Chk1. In addition, this study also demonstrates that Chk1 is essential for IR-induced inhibition of DNA synthesis and the G(2)/M checkpoint. PMID- 12588869 TI - High precision NMR structure and function of the RING-H2 finger domain of EL5, a rice protein whose expression is increased upon exposure to pathogen-derived oligosaccharides. AB - EL5, a RING-H2 finger protein, is rapidly induced by N acetylchitooligosaccharides in rice cell. We expressed the EL5 RING-H2 finger domain in Escherichia coli and determined its structure in solution by NMR spectroscopy. The EL5 RING-H2 finger domain consists of two-stranded beta-sheets (beta1, Ala(147)-Phe(149); beta2, Gly(156)-His(158)), one alpha-helix (Cys(161) Leu(166)), and two large N- and C-terminal loops. It is stabilized by two tetrahedrally coordinated zinc ions. This structure is similar to that of other RING finger domains of proteins of known function. From structural analogies, we inferred that the EL5 RING-H2 finger is a binding domain for ubiquitin conjugating enzyme (E2). The binding site is probably formed by solvent-exposed hydrophobic residues of the N- and C-terminal loops and the alpha-helix. We demonstrated that the fusion protein with EL5-(96-181) and maltose-binding protein (MBP) was polyubiquitinated by incubation with ubiquitin, ubiquitin activating enzyme (E1), and a rice E2 protein, OsUBC5b. This supported the idea that the EL5 RING finger domain is essential for ubiquitin-ligase activity of EL5. By NMR titration experiments, we identified residues that are critical for the interaction between the EL5 RING-H2 finger and OsUBC5b. We conclude that the RING-H2 finger domain of EL5 is the E2 binding site of EL5. PMID- 12588870 TI - Nicotine modulates the expression of a diverse set of genes in the neuronal SH SY5Y cell line. AB - Nicotine exposure can have long lasting effects on nervous system function, some of which must contribute to nicotine dependence. Up-regulation, an increase in numbers of radioligand-binding nicotinic acetylcholine receptors (nAChR), occurs on exposure to nicotine at high concentrations. To determine whether altered gene expression might account for long term changes and up-regulation following nicotine exposure, we assessed effects of 1 h of 1 mm nicotine exposure on alteration of gene expression in the neuron-like SH-SY5Y neuroblastoma clonal line. Repeat and cross-controlled microarray analyses yielded a list of 17 genes from the initially screened approximately 5,000 whose expression was consistently altered following nicotine treatment. Subsequent quantitative, real time reverse transcriptase PCR analyses confirmed altered expression in 14 of 16 genes tested. Further, the general nAChR antagonist, d-tubocurarine, blocked all but two of the observed changes in gene expression, indicating that these changes are dependent on nAChR activation. Use of other antagonists revealed that nAChR subtypes can differentially affect gene expression. The genes affected code for proteins that may be broadly categorized into four groups: transcription factors, protein processing factors, RNA-binding proteins, and plasma membrane-associated proteins. Our results suggest that nicotinic activation of nAChR may have a broad role in affecting cellular physiology through modulating gene expression. PMID- 12588871 TI - Src-mediated RGS16 tyrosine phosphorylation promotes RGS16 stability. AB - The amplitude of signaling evoked by stimulation of G protein-coupled receptors may be controlled in part by the GTPase accelerating activity of the regulator of G protein signaling (RGS) proteins. In turn, subcellular targeting, protein protein interactions, or post-translational modifications such as phosphorylation may shape RGS activity and specificity. We found previously that RGS16 undergoes tyrosine phosphorylation on conserved tyrosine residues in the RGS box. Phosphorylation on Tyr(168) was mediated by the epidermal growth factor receptor (EGFR). We show here that endogenous RGS16 is phosphorylated after epidermal growth factor stimulation of MCF-7 cells. In addition, p60-Src or Lyn kinase phosphorylated recombinant RGS16 in vitro, and RGS16 underwent phosphorylation in the presence of constitutively active Src (Y529F) in EGFR(-) CHO-K1 cells. Blockade of endogenous Src activity by selective inhibitors attenuated RGS16 phosphorylation induced by pervanadate or receptor stimulation. Furthermore, the rate of RGS16 degradation was reduced in cells expressing active Src or treated with pervanadate or a G protein-coupled receptor ligand (CXCL12). Induction of RGS16 tyrosine phosphorylation was associated with increased RGS16 protein levels and enhanced GAP activity in cell membranes. These results suggest that Src mediates RGS16 tyrosine phosphorylation, which may promote RGS16 stability. PMID- 12588872 TI - Mutations in autolytic loop-2 and at Asp554 of human prothrombin that enhance protein C activation by meizothrombin. AB - Thrombin acts on many protein substrates during the hemostatic process. Its specificity for these substrates is modulated through interactions at regions remote from the active site of the thrombin molecule, designated exosites. Exosite interactions can be with the substrate, cofactors such as thrombomodulin, or fragments from prothrombin. The relative activity of alpha-thrombin for fibrinogen is 10 times greater than that for protein C. However, the relative activity of meizothrombin for protein C is 14 times greater than that for fibrinogen. Modulation of thrombin specificity is linked to its Na(+)-binding site and residues in autolytic loop-2 that interact with the Na(+)-binding site. Recombinant prothrombins that yield recombinant meizothrombin (rMT) and rMT des fragment 1 (rMT(desF1)) enable comparisons of the effects of mutations at the Na(+)-binding residue (Asp(554)) and deletion of loop-2 (Glu(466)-Thr(469)) on the relative activity of meizothrombin for several substrates. Hydrolysis of t butoxycarbonyl-VPR-p-nitroanilide by alpha-thrombin, recombinant alpha-thrombin, or rMT(desF1) was almost identical, but that by rMT was only 40% of that by alpha thrombin. Clotting of fibrinogen by rMT and rMT(desF1) was 12-16% of that by alpha-thrombin, as already known. Strikingly, however, although meizothrombins modified by substitution of Asp(554) with either Ala or Leu or by deletion of loop-2 had 6-8 and <1%, respectively, of the clotting activity of alpha-thrombin, the activity of these meizothrombins for protein C was increased to >10 times that of alpha-thrombin. It is proposed that interactions within thrombin that involve autolytic loop-2 and the Na(+)-binding site primarily enhance thrombin action on fibrinogen, but impair thrombin action on protein C. PMID- 12588873 TI - Three-dimensional reconstruction of a light-harvesting complex I-photosystem I (LHCI-PSI) supercomplex from the green alga Chlamydomonas reinhardtii. Insights into light harvesting for PSI. AB - A supercomplex containing the photosystem I (PSI) and chlorophyll a/b light harvesting complex I (LHCI) has been isolated using a His-tagged mutant of Chlamydomonas reinhardtii. This LHCI-PSI supercomplex contained approximately 215 chlorophyll molecules of which 175 were estimated to be chlorophyll a and 40 to be chlorophyll b, based on P700 oxidation and chlorophyll a/b ratio measurements. Its room temperature long wavelength absorption peak was at 680 nm, and it emitted chlorophyll fluorescence maximally at 715 nm (77 K). The LHCI was composed of four or more different types of Lhca polypeptides including Lhca3. No LHCII proteins or other phosphoproteins were detected in the LHCI-PSI supercomplexes suggesting that the cells from which they were isolated were in State 1. Electron microscopy of negatively stained samples followed by image analysis revealed the LHCI-PSI supercomplex to have maximal dimensions of 220 A by 180 A and to be approximately 105 A thick. An averaged top view was used to model in x-ray and electron crystallographic data for PSI and Lhca proteins respectively. We conclude that the supercomplex consists of a PSI reaction center monomer with 11 Lhca proteins arranged along the side where the PSI proteins, PsaK, PsaJ, PsaF, and PsaG are located. The estimated molecular mass for the complex is 700 kDa including the bound chlorophyll molecules. The assignment of 11 Lhca proteins is consistent with a total chlorophyll level of 215 assuming that the PSI reaction center core binds approximately 100 chlorophylls and that each Lhca subunit binds 10 chlorophylls. There was no evidence for oligomerization of Chlamydomonas PSI in contrast to the trimerization of PSI in cyanobacteria. PMID- 12588874 TI - YY1 is regulated by O-linked N-acetylglucosaminylation (O-glcNAcylation). AB - YY1 is a zinc finger DNA-binding transcription factor that influences expression of a wide variety of cellular and viral genes. YY1 is essential for the development of mammalian embryos. It regulates the expression of genes with important functions in DNA replication, protein synthesis, and cellular response to external stimuli during cell growth and differentiation. How YY1 accomplishes such a variety of functions is unknown. Here, we show that a subset of the nuclear YY1 appears to be O-GlcNAcylated regardless of the differentiation status of the cells. We found that glucose strongly stimulates O-linked N acetylglucosaminylation (O-GlcNAcylation) on YY1. Glycosylated YY1 no longer binds the retinoblastoma protein (Rb). Upon dissociation from Rb, the glycosylated YY1 is free to bind DNA. The ability of the O-glycosylation on YY1 to disrupt the complex with Rb leads us to propose that O-glycosylation might have a profound effect on cell cycle transitions that regulate the YY1-Rb heterodimerization and promote the activity of YY1. Our observations provide strong evidence that YY1-regulated transcription is very likely connected to the pathway of glucose metabolism that culminates in the O-GlcNAcylation on YY1, changing its function in transcription. PMID- 12588875 TI - MAPK signaling up-regulates the activity of hypoxia-inducible factors by its effects on p300. AB - Hypoxia-inducible factors (HIF) are a family of heterodimeric transcriptional regulators that play pivotal roles in the regulation of cellular utilization of oxygen and glucose and are essential transcriptional regulators of angiogenesis in solid tumor and ischemic disorders. The transactivation activity of HIF complexes requires the recruitment of p300/CREB-binding protein (CBP) by HIF-1 alpha and HIF-2 alpha that undergo oxygen-dependent degradation. HIF activation in tumors is caused by several factors including mitogen-activated protein kinase (MAPK) signaling. Here we investigated the molecular basis for HIF activation by MAPK. We show that MAPK is required for the transactivation activity of HIF-1 alpha. Furthermore, inhibition of MAPK disrupts the HIF-p300 interaction and suppresses the transactivation activity of p300. Overexpression of MEK1, an upstream MAPK activator, stimulates the transactivation of both p300 and HIF-1 alpha. Interestingly, the C-terminal transactivation domain of HIF-1 alpha is not a direct substrate of MAPK, and HIF-1 alpha phosphorylation is not required for HIF-CAD/p300 interaction. Taken together, our data suggest that MAPK signaling facilitates HIF activation through p300/CBP. PMID- 12588876 TI - Structural analysis of Bacillus subtilis SPP1 phage helicase loader protein G39P. AB - The Bacillus subtilis SPP1 phage-encoded protein G39P is a loader and inhibitor of the phage G40P replicative helicase involved in the initiation of DNA replication. We have carried out a full x-ray crystallographic and preliminary NMR analysis of G39P and functional studies of the protein, including assays for helicase binding by a number of truncated mutant forms, in an effort to improve our understanding of how it both interacts with the helicase and with the phage replisome organizer, G38P. Our structural analyses reveal that G39P has a completely unexpected bipartite structure comprising a folded N-terminal domain and an essentially unfolded C-terminal domain. Although G39P has been shown to bind its G40P target with a 6:6 stoichiometry, our crystal structure and other biophysical characterization data reveal that the protein probably exists predominantly as a monomer in solution. The G39P protein is proteolytically sensitive, and our binding assays show that the C-terminal domain is essential for helicase interaction and that removal of just the 14 C-terminal residues abolishes interaction with the helicase in vitro. We propose a number of possible scenarios in which the flexibility of the C-terminal domain of G39P and its proteolytic sensitivity may have important roles for the function of G39P in vivo that are consistent with other data on SPP1 phage DNA replication. PMID- 12588877 TI - Oxidative stress and diabetic neuropathy: a new understanding of an old problem. PMID- 12588878 TI - Insulin's effect on glucose production: direct or indirect? PMID- 12588879 TI - Insights into channel function via channel dysfunction. PMID- 12588880 TI - The role of uncoupling protein 3 in human physiology. PMID- 12588881 TI - RGS2: a "turn-off" in hypertension. PMID- 12588882 TI - Hypertension and prolonged vasoconstrictor signaling in RGS2-deficient mice. AB - Signaling by hormones and neurotransmitters that activate G protein-coupled receptors (GPCRs) maintains blood pressure within the normal range despite large changes in cardiac output that can occur within seconds. This implies that blood pressure regulation requires precise kinetic control of GPCR signaling. To test this hypothesis, we analyzed mice deficient in RGS2, a GTPase-activating protein that greatly accelerates the deactivation rate of heterotrimeric G proteins in vitro. Both rgs2+/- and rgs2-/- mice exhibited a strong hypertensive phenotype, renovascular abnormalities, persistent constriction of the resistance vasculature, and prolonged response of the vasculature to vasoconstrictors in vivo. Analysis of P2Y receptor-mediated Ca2+ signaling in vascular smooth muscle cells in vitro indicated that loss of RGS2 increased agonist potency and efficacy and slowed the kinetics of signal termination. These results establish that abnormally prolonged signaling by G protein-coupled vasoconstrictor receptors can contribute to the onset of hypertension, and they suggest that genetic defects affecting the function or expression of RGS2 may be novel risk factors for development of hypertension in humans. PMID- 12588883 TI - Inhibition of adipogenesis and development of glucose intolerance by soluble preadipocyte factor-1 (Pref-1). AB - Preadipocyte factor-1 (Pref-1) is a transmembrane protein highly expressed in preadipocytes. Pref-1 expression is, however, completely abolished in adipocytes. The extracellular domain of Pref-1 undergoes two proteolytic cleavage events that generate 50 and 25 kDa soluble products. To understand the function of Pref-1, we generated transgenic mice that express the full ectodomain corresponding to the large cleavage product of Pref-1 fused to human immunoglobulin-gamma constant region. Mice expressing the Pref-1/hFc transgene in adipose tissue, driven by the adipocyte fatty acid-binding protein (aP2, also known as aFABP) promoter, showed a substantial decrease in total fat pad weight. Moreover, adipose tissue from transgenic mice showed reduced expression of adipocyte markers and adipocyte secreted factors, including leptin and adiponectin, whereas the preadipocyte marker Pref-1 was increased. Pref-1 transgenic mice with a substantial, but not complete, loss of adipose tissue exhibited hypertriglyceridemia, impaired glucose tolerance, and decreased insulin sensitivity. Mice expressing the Pref-1/hFc transgene exclusively in liver under the control of the albumin promoter also showed a decrease in adipose mass and adipocyte marker expression, suggesting an endocrine mode of action of Pref-1. These findings demonstrate the inhibition of adipogenesis by Pref-1 in vivo and the resulting impairment of adipocyte function that leads to the development of metabolic abnormalities. PMID- 12588884 TI - Insulin signaling is required for insulin's direct and indirect action on hepatic glucose production. AB - We and others have suggested that insulin predominantly acts indirectly to inhibit hepatic glucose production (HGP) via suppression of gluconeogenic precursors, FFAs, and glucagon. To test that hypothesis, we performed high-dose hyperinsulinemic-euglycemic clamps using [3-(3)H]-glucose in liver-specific insulin receptor knockout (LIRKO) mice, LIRKO mice treated with streptozotocin (LIRKO+STZ), and controls. In LIRKO mice, fasted glucose was normal, but insulin levels were elevated tenfold. STZ treatment reduced insulinemia by 60% with resulting hyperglycemia. Interestingly, basal HGP was similar in all three groups. During the clamp, HGP was suppressed by 82 +/- 17% in controls, but was not suppressed in either LIRKO or LIRKO+STZ mice. Glucose infusion and utilization were impaired ( approximately 50%) in LIRKO and LIRKO+STZ mice versus controls. Insulin suppressed FFAs similarly in all groups ( approximately 46%). Glucagon was not significantly suppressed during the clamp. Thus, in LIRKO mice, (a) high-dose insulin fails to suppress HGP indicating that both direct and indirect effects of insulin require an intact insulin-signaling pathway in the liver; (b) primary hepatic insulin resistance leads to hyperinsulinemia and secondary extrahepatic insulin resistance; and (c) lowering insulin levels with STZ tended to improve extrahepatic insulin sensitivity but failed to reveal the previously postulated indirect role of insulin in suppressing HGP. PMID- 12588885 TI - The suppressor of cytokine signaling-1 (SOCS1) is a novel therapeutic target for enterovirus-induced cardiac injury. AB - Enteroviral infections of the heart are among the most commonly identified causes of acute myocarditis in children and adults and have been implicated in dilated cardiomyopathy. Although there is considerable information regarding the cellular immune response in myocarditis, little is known about innate signaling mechanisms within the infected cardiac myocyte that contribute to the host defense against viral infection. Here we show the essential role of Janus kinase (JAK) signaling in cardiac myocyte antiviral defense and a negative role of an intrinsic JAK inhibitor, the suppressor of cytokine signaling (SOCS), in the early disease process. Cardiac myocyte-specific transgenic expression of SOCS1 inhibited enterovirus-induced signaling of JAK and the signal transducers and activators of transcription (STAT), with accompanying increases in viral replication, cardiomyopathy, and mortality in coxsackievirus-infected mice. Furthermore, the inhibition of SOCS in the cardiac myocyte through adeno-associated virus-mediated (AAV-mediated) expression of a dominant-negative SOCS1 increased the myocyte resistance to the acute cardiac injury caused by enteroviral infection. These results indicate that strategies directed at inhibition of SOCS in the heart and perhaps other organs can augment the host-cell antiviral system, thus preventing viral-mediated end-organ damage during the early stages of infection. PMID- 12588886 TI - Increased uncoupling protein 3 content does not affect mitochondrial function in human skeletal muscle in vivo. AB - Phosphocreatine (PCr) resynthesis rate following intense anoxic contraction can be used as a sensitive index of in vivo mitochondrial function. We examined the effect of a diet-induced increase in uncoupling protein 3 (UCP3) expression on postexercise PCr resynthesis in skeletal muscle. Nine healthy male volunteers undertook 20 one-legged maximal voluntary contractions with limb blood flow occluded to deplete muscle PCr stores. Exercise was performed following 7 days consumption of low-fat (LF) or high-fat (HF) diets. Immediately following exercise, blood flow was reinstated, and muscle was sampled after 20, 60, and 120 seconds of recovery. Mitochondrial coupling was assessed by determining the rate of PCr resynthesis during recovery. The HF diet increased UCP3 protein content by approximately 44% compared with the LF diet. However, this HF diet-induced increase in UCP3 expression was not associated with any changes in the rate of muscle PCr resynthesis during conditions of maximal flux through oxidative phosphorylation. Muscle acetylcarnitine, free-creatine, and lactate concentrations during recovery were unaffected by the HF diet. Taken together, our findings demonstrate that increasing muscle UCP3 expression does not diminish the rate of PCr resynthesis, allowing us to conclude that the primary role of UCP3 in humans is not uncoupling. PMID- 12588887 TI - Enhanced growth of tumors in SPARC null mice is associated with changes in the ECM. AB - SPARC, a 32-kDa glycoprotein, participates in the regulation of morphogenesis and cellular differentiation through its modulation of cell-matrix interactions. Major functions defined for SPARC in vitro are de-adhesion and antiproliferation. In vivo, SPARC is restricted in its expression to remodeling tissues, including pathologies such as cancer. However, the function of endogenous SPARC in tumor growth and progression is not known. Here, we report that implanted tumors grew more rapidly in mice lacking SPARC. We observed that tumors grown in SPARC null mice showed alterations in the production and organization of ECM components and a decrease in the infiltration of macrophages. However, there was no change in the levels of angiogenic growth factors in comparison to tumors grown in wild type mice, although there was a statistically significant difference in total vascular area. Whereas SPARC did inhibit the growth of tumor cells in vitro, it did not have a demonstrable effect on the proliferation or apoptosis of tumor cells in vivo. These data indicate that host-derived SPARC is important for the appropriate organization of the ECM in response to implanted tumors and highlight the importance of the ECM in regulating tumor growth. PMID- 12588888 TI - Mutation causing severe myasthenia reveals functional asymmetry of AChR signature cystine loops in agonist binding and gating. AB - We describe a highly disabling congenital myasthenic syndrome (CMS) associated with rapidly decaying, low-amplitude synaptic currents, and trace its cause to a valine to leucine mutation in the signature cystine loop (cys-loop) of the AChR alpha subunit. The recently solved crystal structure of an ACh-binding protein places the cys-loop at the junction between the extracellular ligand-binding and transmembrane domains where it may couple agonist binding to channel gating. We therefore analyzed the kinetics of ACh-induced single-channel currents to identify elementary steps in the receptor activation mechanism altered by the alphaV132L mutation. The analysis reveals that alphaV132L markedly impairs ACh binding to receptors in the resting closed state, decreasing binding affinity for the second binding step 30-fold, but attenuates gating efficiency only about twofold. By contrast, mutation of the equivalent valine residue in the delta subunit impairs channel gating approximately fourfold with little effect on ACh binding, while corresponding mutations in the beta and epsilon subunits are without effect. The unique functional contribution of the alpha subunit cys-loop likely owes to its direct connection via a beta strand to alphaW149 at the center of the ligand-binding domain. The overall findings reveal functional asymmetry between cys-loops of the different AChR subunits in contributing to ACh binding and channel gating. PMID- 12588889 TI - Therapeutic efficacy of sonic hedgehog protein in experimental diabetic neuropathy. AB - Hedgehog proteins modulate development and patterning of the embryonic nervous system. As expression of desert hedgehog and the hedgehog receptor, patched-1, persist in the postnatal and adult peripheral nerves, the hedgehog pathway may have a role in maturation and maintenance of the peripheral nervous system in normal and disease states. We measured desert hedgehog expression in the peripheral nerve of maturing diabetic rats and found that diabetes caused a significant reduction in desert hedgehog mRNA. Treating diabetic rats with a sonic hedgehog-IgG fusion protein fully restored motor- and sensory-nerve conduction velocities and maintained the axonal caliber of large myelinated fibers. Diabetes-induced deficits in retrograde transport of nerve growth factor and sciatic-nerve levels of calcitonin gene-related product and neuropeptide Y were also ameliorated by treatment with the sonic hedgehog-IgG fusion protein, as was thermal hypoalgesia in the paw. These studies implicate disruption of normal hedgehog function in the etiology of diabetes-induced peripheral-nerve dysfunction and indicate that delivery of exogenous hedgehog proteins may have therapeutic potential for the treatment of diabetic neuropathy. PMID- 12588890 TI - Albumin stimulates interleukin-8 expression in proximal tubular epithelial cells in vitro and in vivo. AB - Renal tubulointerstitial injury is characterized by inflammatory cell infiltrate; however, the stimuli for leukocyte recruitment are not fully understood. IL-8 is a potent chemokine produced by proximal tubular epithelial cells (PTECs). Whether nephrotic proteins stimulate tubular IL-8 expression remains unknown. Acute exposure of human PTECs to albumin induced IL-8 gene and protein expression time- and dose-dependently. Apical albumin predominantly stimulated basolateral IL-8 secretion. Electrophoretic mobility shift assay demonstrated nuclear translocation of NF-kappaB, and the p65/p50 subunits were activated. NF-kappaB activation and IL-8 secretion were attenuated by the NF-kappaB inhibitors pyrrolidine dithiocarbamate and cell-permeable peptide. Albumin upregulated intracellular reactive oxygen species (ROS) generation, while exogenous H2O2 stimulated NF-kappaB translocation and IL-8 secretion. Albumin-induced ROS generation, NF-kappaB activation, and IL-8 secretion were endocytosis- and PKC dependent as these downstream events were abrogated by the PI3K inhibitors LY294002 and wortmannin, and the PKC inhibitors GF109203X and staurosporin, respectively. In vivo, IL-8 mRNA expression was localized by in situ hybridization to the proximal tubules in nephrotic kidney tissues. The intensity of IL-8 immunostaining was higher in nephrotic than non-nephrotic subjects. In conclusion, albumin is a strong stimulus for tubular IL-8 expression, which occurs via NF-kappaB-dependent pathways through PKC activation and ROS generation. PMID- 12588891 TI - Identification of Hodgkin and Reed-Sternberg cell-specific genes by gene expression profiling. AB - Hodgkin lymphoma (HL) is a malignancy of unknown pathogenesis. The malignant Hodgkin and Reed/Sternberg (HRS) cells derive from germinal center B cells (or rarely, T cells) but have a heterogeneous and largely uncharacterized phenotype. Using microarrays, we compared the gene expression profile of four HL cell lines with profiles of the main B cell subsets and B cell non-HLs to find out whether HRS cells, despite their described heterogeneity, show a distinct gene expression, to study their relationship to other normal and malignant B cells, and to identify genes aberrantly or overexpressed by HRS cells. The HL lines indeed clustered as a distinct entity, irrespective of their B or T cell derivation, and their gene expression was most similar to that of EBV-transformed B cells and cell lines derived from diffuse large cell lymphomas showing features of in vitro-activated B cells. Twenty-seven genes, most of which were previously unknown to be expressed by HRS cells, showed aberrant expression specifically in these cells, e.g., the transcription factors GATA-3, ABF1, EAR3, and Nrf3. For five genes, expression in primary HRS cells was confirmed. The newly identified HL-specific genes may play important roles in the pathogenesis of HL, potentially represent novel diagnostic markers, and can be considered for therapeutic targeting. PMID- 12588892 TI - Histone deacetylase inhibitors modulate renal disease in the MRL-lpr/lpr mouse. AB - Studies in human systemic lupus erythematosus (SLE) suggest a possible role for histone deacetylases (HDACs) in skewed gene expression and disease pathogenesis. We used the MRL-lpr/lpr murine model of lupus to demonstrate that HDACs play a key role in the heightened levels of both Th1 and Th2 cytokine expression that contribute to disease. The availability of specific HDAC inhibitors (HDIs) such as trichostatin A (TSA) and suberonylanilide hydroxamic acid (SAHA) permits the study of the role of HDACs in gene regulation. Our results indicate that HDIs downregulate IL-12, IFN-gamma, IL-6, and IL-10 mRNA and protein levels in MRL lpr/lpr splenocytes. This effect on gene transcription is associated with an increased accumulation of acetylated histones H3 and H4 in total cellular chromatin. To elucidate the in vivo effects of TSA on lupuslike disease, we treated MRL-lpr/lpr mice with TSA (0.5 mg/kg/d) for 5 weeks. Compared with vehicle-treated control mice, TSA-treated mice exhibited a significant reduction in proteinuria, glomerulonephritis, and spleen weight. Taken together, these findings suggest that increased expression of HDACs leading to an altered state of histone acetylation may be of pathologic significance in MRL-lpr/lpr mice. In addition, TSA or other HDIs may have therapeutic benefit in the treatment of SLE. PMID- 12588893 TI - Regulation of Stat3 nuclear export. AB - Stat3 is the most pleiotropic member of the signal transducer and activator of transcription (STAT) family of transcription factors and mediates pivotal responses for the cytokine family. In resting cells, STATs, including Stat3, reside largely in the cytoplasm. Upon cytokine stimulation, they rapidly translocate to the nucleus, where they promote the expression of target genes. During the subsequent period of signal decay they are re-exported back to the cytoplasm in preparation for the next round of signaling. This process of nuclear export can be blocked by the fungal toxin leptomycin B (LMB). In contrast to what appears to be the case for Stat1, LMB treatment not only blocks the poststimulation export of Stat3 from the nucleus back to the cytoplasm, but also promotes the nuclear accumulation of Stat3 in resting cells. Remarkably, the LMB dependent nuclear accumulation of Stat3 in resting cells is independent of tyrosine phosphorylation, highlighting the existence of a "basal" signaling pathway. Subsequent studies identified three nuclear export signal (NES) elements. Two of these elements, Stat3(306-318) and Stat3(404-414), corresponded to those recently identified in Stat1, and a third, Stat3(524-535), is novel. Stat3(306-318) appears to be important in the rapid nuclear export seen after stimulation (poststimulation export), whereas the Stat3(404-414) and Stat3(524 535) play a more important role in regulating basal nuclear export. In summary, these studies indicate that the process of Stat3 nuclear export is dependent on multiple NES elements. PMID- 12588894 TI - Mechanisms of the cerebrovascular response to apnoea in humans. AB - We measured ventilation, arterial O2 saturation, end-tidal CO2 (PET,CO2), blood pressure (intra-arterial catheter or photoelectric plethysmograph), and flow velocity in the middle cerebral artery (CFV) (pulsed Doppler ultrasound) in 17 healthy awake subjects while they performed 20 s breath holds under control conditions and during ganglionic blockade (intravenous trimethaphan, 4.4 +/- 1.1 mg min-1 (mean +/- S.D.)). Under control conditions, breath holding caused increases in PET,CO2 (7 +/- 1 mmHg) and in mean arterial pressure (MAP) (15 +/- 2 mmHg). A transient hyperventilation (PET,CO2 -7 +/- 1 mmHg vs. baseline) occurred post-apnoea. CFV increased during apnoeas (by 42 +/- 3 %) and decreased below baseline (by 20 +/- 2 %) during post-apnoea hyperventilation. In the post-apnoea recovery period, CFV returned to baseline in 45 +/- 4 s. The post-apnoea decrease in CFV did not occur when hyperventilation was prevented. During ganglionic blockade, which abolished the increase in MAP, apnoea-induced increases in CFV were partially attenuated (by 26 +/- 2 %). Increases in PET,CO2 and decreases in oxyhaemoglobin saturation (Sa,O2) (by 2 +/- 1 %) during breath holds were identical in the intact and blocked conditions. Ganglionic blockade had no effect on the slope of the CFV response to hypocapnia but it reduced the CFV response to hypercapnia (by 17 +/- 5 %). We attribute this effect to abolition of the hypercapnia-induced increase in MAP. Peak increases in CFV during 20 s Mueller manoeuvres (40 +/- 3 %) were the same as control breath holds, despite a 15 mmHg initial, transient decrease in MAP. Hyperoxia also had no effect on the apnoea induced increase in CFV (40 +/- 4 %). We conclude that apnoea-induced fluctuations in CFV were caused primarily by increases and decreases in arterial partial pressure of CO2 (Pa,CO2) and that sympathetic nervous system activity was not required for either the initiation or the maintenance of the cerebrovascular response to hyper- and hypocapnia. Increased MAP or other unknown influences of autonomic activation on the cerebral circulation played a smaller but significant role in the apnoea-induced increase in CFV; however, negative intrathoracic pressure and the small amount of oxyhaemoglobin desaturation caused by 20 s apnoea did not affect CFV. PMID- 12588895 TI - Effect of ageing on the electrical and mechanical properties of human soleus motor units activated by the H reflex and M wave. AB - This study was designed to investigate the effect of ageing on the mechanical and electromyographic (EMG) characteristics of the soleus motor units (MUs) activated by the maximal Hoffmann reflex (Hmax) and by the direct muscle compound action potential (Mmax). Eleven young (mean age 25 +/- 4 years) and ten elderly (mean age 73 +/- 5 years) males took part in this investigation. The senior group presented lower amplitudes of Mmax (-57 %, P < 0.001) and Hmax (-68 %, P < 0.001) waves compared to the younger population. These were associated with a depression of relative twitch torque of the plantar flexors. The average values of the Hmax/Mmax ratio did not statistically differ between the two populations, despite a tendency for lower values (~23 %) in the senior group. However, the older adults showed a greater relative amplitude of the sub-maximal M wave evoked at Hmax (MatHmax) than did the younger males (young 5 % vs. elderly 29 % of the Mmax, P < 0.01). This finding suggests an increased homogeneity between the excitability threshold of sensory and motor axons. The twitch torque at Hmax (PtH M) was subsequently calculated by subtraction from the total twitch torque of the mechanical contamination associated with MatHmax. The resulting PtH-M was significantly lower in the elderly (-59 %, P < 0.001). Despite a discrepancy of 20 % between the two groups, the mechanical ratio (PtH-M/PtM; PtM, twitch tension related to the Mmax compound action potential), like the EMG ratio, did not statistically differ between the young and older individuals. Nevertheless, the senior subjects exhibited a higher twitch/EMG ratio for the reflexively activated MUs (PtH-M/Hmax) than the younger individuals (+40 %, P < 0.05). This finding suggests an on-going neuromuscular remodelling, resulting in an increased innervation ratio. The neural rearrangement may be viewed as a compensatory adaptation of the motor system to preserve the mechanical efficiency of the surviving MUs, despite the age-related impairment of the segmental reflex system. This phenomenon is confirmed by the maintenance, with senescence, of the approximately constant values of the twitch/EMG ratio for the entire motor pool (PtM/Mmax). PMID- 12588896 TI - Negative charges in the DIII-DIV linker of human skeletal muscle Na+ channels regulate deactivation gating. AB - Charge reversing, neutralizing and substituting mutations at D1309 and EE1314,15 in the DIII-DIV linker of the human skeletal muscle sodium channel hNav1.4 were constructed and expressed in Xenopus oocytes. The effects of these mutations on conductance, inactivation and deactivation were determined using on-cell macropatches. D1309R caused a depolarizing shift of the conductance-voltage (g(V)) curve and increased the apparent valency of activation. D1309R and EE1314,15RR increased time to peak activation. D1309R caused a depolarizing shift of the steady-state fast inactivation curve, whereas EE1314,15RR produced a hyperpolarizing shift and decreased the apparent valency. Charge reversal at either D1309 or EE1314,15 slowed open-state fast inactivation and accelerated closed-state fast inactivation. D1309R accelerated recovery from fast inactivation, whereas EE1314,15RR and EE1314,15QQ slowed recovery. Deactivation from the inactivated state was determined by the delay in the onset to recovery from fast inactivation. Recovery delay was abbreviated for D1309R but was prolonged for EE1314,15RR and EE1314,15QQ. Open-state deactivation was determined from the time constant of the decay (tau D) of tail currents. tau D was slowed by D1309R, D1309E, EE1314,15RR and EE1314,15QQ. Our findings suggest an important role in deactivation gating in hNav1.4 for the negative cluster of charge at EE1314,15. These and previous findings suggest that clusters of negatively and positively charged residues in the hNav1.4 DIII-DIV linker differentially regulate the kinetics of fast inactivation. PMID- 12588897 TI - Developmental changes in the expression of potassium currents of embryonic, neonatal and mature mouse inner hair cells. AB - Developmental changes in electrophysiological membrane properties of mouse cochlear inner hair cells (IHCs) were studied from just after terminal differentiation up to functional maturity. As early as embryonic day 14.5 (E14.5) newly differentiated IHCs express a very small outward K+ current that is largely insensitive to 4-aminopyridine (4-AP). One day later the inward rectifier, IK1, is first observed. These immature cells initially exhibit only slow graded voltage responses under current clamp. From E17.5 spontaneous action potentials occur. During the first week of postnatal development, the outward K+ current steadily increases in size and a progressively larger fraction of the current is sensitive to 4-AP. During the second postnatal week, the activation of the 4-AP sensitive current, by now contributing about half of the outward K+ current, shifts in the hyperpolarizing direction. Together with an increase in size of IK1, this hyperpolarizes the cell, thus inhibiting the spontaneous spike activity, although spikes could still be evoked upon depolarizing current injection. Starting at about the onset of hearing (postnatal day 12, P12) immature IHCs make the final steps towards fully functional sensory receptors with fast graded voltage responses. This is achieved mainly by the expression of the large-conductance Ca2+-activated K+ current IK,f, but also of a current indistinguishable from the negatively activating IK,n previously described in mature outer hair cells (OHCs). The 4-AP-sensitive current continues to increase after the onset of hearing to form the major part of the mature delayed rectifier, IK,s. By P20 IHCs appear mature in terms of their complement of K+ conductances. PMID- 12588898 TI - Mitochondrial Ca2+ uptake prevents desynchronization of quantal release and minimizes depletion during repetitive stimulation of mouse motor nerve terminals. AB - We investigated how inhibition of mitochondrial Ca2+ uptake affects transmitter release from mouse motor terminals during brief trains of action potentials (500 at 50 Hz) in physiological bath [Ca2+]. When mitochondrial Ca2+ uptake was inhibited by depolarizing mitochondria with antimycin A1 or carbonyl cyanide m chlorophenyl-hydrazone, the stimulation-induced increase in cytosolic [Ca2+] was greater (> 10 microM, compared to < or = 1 microM in control solution), the quantal content of the endplate potential (EPP) depressed more rapidly (approximately 84 % depression compared to approximately 8 % in controls), and asynchronous release during the stimulus train reached higher frequencies (peak rates of approximately 6000 s-1 compared to approximately 75 s-1 in controls). These effects of mitochondrial depolarization were not accompanied by a significant change in EPP quantal content or the rate of asynchronous release during 1 Hz stimulation, and were not seen in oligomycin, which blocks mitochondrial ATP synthesis without depolarizing mitochondria. Inhibition of endoplasmic reticular Ca2+ uptake with cyclopiazonic acid also had little effect on stimulation-induced changes in cytosolic [Ca2+] or EPP amplitude. We hypothesize that the high rate of asynchronous release evoked by stimulation during mitochondrial depolarization was produced by the elevation of cytosolic [Ca2+], and contributed to the accelerated depression of phasic release by reducing the availability of releasable vesicles. During mitochondrial depolarization, the post-tetanic potentiation of the EPP observed under control conditions was replaced by a post-tetanic depression with a slow time course of recovery. Thus, mitochondrial Ca2+ uptake is essential for sustaining phasic release, and thus neuromuscular transmission, during and following tetanic stimulation. PMID- 12588899 TI - Phosphorylation of protein kinase C sites in NBD1 and the R domain control CFTR channel activation by PKA. AB - Activation of the cystic fibrosis transmembrane conductance regulator (CFTR) channel by protein kinase A (PKA) is enhanced by protein kinase C (PKC). However, the mechanism of modulation is not known and it remains uncertain whether PKC acts directly on CFTR or through phosphorylation of an ancillary protein. Using excised patches that had been pre-treated with phosphatases, we found that PKC exposure results in much larger PKA-activated currents and shifts the PKA concentration dependence. To examine if these effects are mediated by direct PKC phosphorylation of CFTR, a mutant was constructed in which serines or threonines at nine PKC consensus sequences on CFTR were replaced by alanines (i.e. the '9CA' mutant T582A/T604A/S641A/T682A/S686A/S707A/S790A/T791A/S809A). In excised patches, 9CA channels had greatly reduced responses to PKA (i.e. 5-10 % that of wild-type), which were not enhanced by PKC pre-treatment, although the mutant channels were still functional according to iodide efflux assays. Stimulation of iodide efflux by chlorophenylthio-cAMP (cpt-cAMP) was delayed in cells expressing 9CA channels, and a similar delay was observed when cells expressing wild-type CFTR were treated with the PKC inhibitor chelerythrine. This suggests that weak activation by PKA in excised patches and slow stimulation of iodide efflux from intact cells are specifically due to the loss of PKC phosphorylation. Finally, PKC caused a slight activation of wild-type channels when added to excised patches after phosphatase pre-treatment but had no effect on the mutant. We conclude that direct phosphorylation of CFTR at one or more of the nine sites mutated in 9CA is required for both the partial activation by PKC and for its modulation of CFTR responses to PKA. PMID- 12588900 TI - Multiple modes of GABAergic inhibition of rat cerebellar granule cells. AB - Cerebellar granule cells are inhibited phasically by GABA released synaptically from Golgi cells, but are inhibited more powerfully by tonic activity of high affinity alpha 6 subunit-containing GABAA receptors. During development the tonic activity is generated by the accumulation of GABA released by action potentials, but in the adult the tonic activity is independent of action potentials. Here we show that in adult rats the tonic activation of GABAA receptors is produced by non-vesicular transmitter release and is reduced by the activity of GAT-1 and GAT 3 GABA transporters, demonstrating that alterations of GABA uptake will modulate information flow through granule cells. Acetylcholine (ACh) evokes a large Ca2+ dependent but action potential-independent release of GABA, which activates alpha 6 subunit-containing GABAA receptors. These data show that three separate modes of transmitter release can activate GABAA receptors in adult cerebellar granule cells: action potential-evoked exocytotic GABA release, non-vesicular release, and ACh-evoked Ca2+-dependent release independent of action potentials. The relative magnitudes of the inhibitory charge transfers generated by action potential-evoked release (during high frequency stimulation of the mossy fibres), tonic inhibition and superfused ACh are 1:3:12, indicating that tonic and ACh mediated inhibition may play a major role in regulating granule cell firing. PMID- 12588901 TI - Neurosteroid regulation of oxytocin and vasopressin release from the rat supraoptic nucleus. AB - In adult rats somato-dendritic release of oxytocin and vasopressin from magnocellular neurones in the supraoptic nucleus of the hypothalamus has important autoregulatory actions on the neuronal electrical activity, and in neonatal rats it plays a role in the development of dendritic arborisation. In the adult, oxytocin effects are modulated by allopregnanolone via an interaction with inhibitory GABAA receptors. This study examined the effects of allopregnanolone, progesterone and 17beta-oestradiol on oxytocin and vasopressin release from intact isolated supraoptic nuclei and from the neurophypophyses in rats of differing ages. In supraoptic nuclei from rats of 3-4 weeks old or less, all three neurosteroids induced oxytocin release from the isolated supraoptic nucleus, but only allopregnanolone induced significant release of vasopressin. Surprisingly, in these very young rats, allopregnanolone-induced oxytocin release was inhibited by GABAA receptor antagonists as well as by an oxytocin receptor antagonist. By contrast, in supraoptic nuclei from adult rats allopregnanolone induced oxytocin release was much smaller, and was enhanced in the presence of bicuculline. The GABAA receptor agonist muscimol also induced oxytocin release from supraoptic nuclei in young rats, but had no effect in adult rats. Oxytocin cells isolated from young rats showed an increase in [Ca2+]i in response to both allopregnanolone and muscimol. Allopregnanolone had no effect on [Ca2+]i or on the release of oxytocin or vasopressin from neurohypophysial axon terminals in either young or old rats. We conclude that, in very young rats, (i) neurosteroids induce oxytocin release from the supraoptic nucleus by a mechanism that partly depends on the presence of GABA, which in young rats is depolarising to oxytocin cells, and which also partly depends upon endogenous oxytocin, and (ii) the effect of allopregnanolone upon oxytocin release changes with age, as the functional activity of GABAA receptors changes from excitation to inhibition of oxytocin cells. PMID- 12588902 TI - alpha1- and alpha2-adrenergic vasoconstriction is blunted in contracting human muscle. AB - Sympathetic vasoconstriction is blunted in the vascular beds of contracting skeletal muscles. We sought to determine whether this blunted vasoconstriction is specific for post-junctional alpha1- or alpha2-adrenergic receptors. We measured forearm blood flow (Doppler ultrasound) and calculated the vascular conductance (FVC) responses to brachial artery infusions of tyramine (which evokes endogenous noradrenaline release), phenylephrine (an alpha1 agonist) and clonidine (an alpha2 agonist) in 10 healthy men during rhythmic handgrip exercise (10-15 % of maximum) and during a control non-exercise vasodilator condition (intra-arterial adenosine). Steady-state FVC during exercise and adenosine was similar in all trials (range: 243-272 and 234-263 ml min-1 (100 mmHg)-1, respectively; P > 0.5). During exercise the percentage reductions in FVC in response to tyramine (-24 +/- 7 vs. -55 +/- 6 %), phenylephrine (-12 +/- 8 vs. -37 +/- 8 %) and clonidine (-17 +/- 6 vs. -49 +/- 4 %) were significantly less compared with adenosine (all P < 0.05). The magnitude of the blunted vasoconstrictor responses was similar for both receptor subtypes. These findings are in contrast to those from studies in animals demonstrating that alpha2-adrenergic receptor-mediated vasoconstrictor responses are much more sensitive to contraction-induced inhibition than alpha1 mediated responses. We conclude that vasoconstrictor responses mediated via both post-junctional alpha1- and alpha2-adrenergic receptors are blunted in contracting human skeletal muscles. PMID- 12588903 TI - A second mechanism of increase of cerebellar hypermetria in humans. AB - So far, there is only one procedure known to increase hypermetria in cerebellar patients. Facing an increased inertia of the moving limb, patients presenting a lesion of the lateral cerebellum are able to increase appropriately the intensity of the agonist electromyographic (EMG) activity (the launching force), but are unable to adapt the intensity of the antagonist activity (the braking force). As a result, hypermetria is larger when the inertial load is artificially increased. Recent studies have demonstrated that hyperventilation increases hypermetria in patients presenting a spinocerebellar ataxia type 6 (SCA 6), a disorder associated with polyglutamine expansions in the alpha1A-voltage-dependent calcium channel. The mechanism of this increase of hypermetria has not been identified so far. In the present work, we combined kinematic, EMG and transcranial Doppler studies to understand the effects of hyperventilation on fast goal-directed movements in patients presenting a SCA 6. Both in the normal mechanical state and after increasing the inertial load of the moving hand, hyperventilation induced an increase of hypermetria. Hyperventilation increased the delay of the onset latency of the antagonist EMG activity and decreased the rate of rise of both the agonist and the antagonist EMG activities. Hyperventilation induced a marked decrease in cerebral blood flow velocities. The mechanism of this provocative test is original and is distinct from the mechanism of the load-induced increase of hypermetria. PMID- 12588904 TI - Patch-clamp analysis in canine cardiac Purkinje cells of a novel sodium component in the pacemaker range. AB - A putative Na+ component playing a role in the initiation and maintenance of spontaneous discharge in Purkinje fibres was studied by means of the whole-cell patch-clamp technique in canine cardiac single Purkinje cells. In 4 mM [K+]o, during depolarising clamp steps, a slowly inactivating current appeared at approximately -58 mV, negative to the threshold for the fast Na+ current (INa; approximately -50 mV). During depolarising ramps, the current underwent inward rectification with a negative slope region that began at approximately -60 mV. The current underlying the negative slope increased during faster ramps, decreased as a function of time when the initial depolarising ramp was over, decreased during depolarisations positive to approximately -35 mV and was much larger than the current during the symmetrical repolarising ramp. Increasing biphasic ('oscillatory') voltage ramps required much smaller currents at a holding potential (Vh) of -60 mV than at -80 mV and were associated with a marked decrease in slope conductance. At Vh -50/-40 mV, the oscillatory ramp currents and superimposed pulse currents reversed direction. The negative slope in the I-V relation as well as the change in current direction at -50/-40 mV were markedly reduced by tetrodotoxin (15 microM) and lidocaine (lignocaine, 100 microM) and therefore are due to a slowly inactivating Na+ current, labelled here INa3. Lower [K+]o (2.7 mM) reduced the steady state slope conductance as well as the current in the diastolic range, and increased as well as shifted INa3 in a negative direction. High [K+]o had the opposite effects. Cs+ (2 mM) and Ba2+ (2 mM) reduced the initial current during depolarising ramps but not INa3. In current clamp mode, current-induced voltage oscillations elicited action potentials through a gradual transition between diastolic depolarisation and upstroke, consistent with the activation of INa3. Thus, the initiation and maintenance of spontaneous discharge in Purkinje strands appear to involve a voltage- and K+ dependent decrease in K+ conductance as well as the activation of a voltage- and time-dependent inward Na+ current (INa3) with slow inactivation kinetics. PMID- 12588905 TI - Letter from Cuba. PMID- 12588906 TI - Vigabatrin, tiagabine, and visual fields. PMID- 12588907 TI - Quality of life in motor neurone disease--towards a more practical assessment tool? PMID- 12588908 TI - Objective measures for the progression of Parkinson's disease. PMID- 12588910 TI - Vestibular reflexes and positional manoeuvres. AB - Dizziness and vertigo are some of the more frequently encountered symptoms in neurology clinics. In turn, one of the most common causes of vertigo is benign paroxysmal positional vertigo (BPPV), accounting for a quarter of all patients with dizziness and vertigo. Reviewing the value of the positional manoeuvres available is relevant, particularly in the light of the efficient treatments available for BPPV. In this article I will deal with positional manoeuvres first, and then with how vestibulo-ocular reflexes (VOR) can be tested in the clinic. I will not discuss VOR suppression assessment. PMID- 12588912 TI - Discrete and dynamic scaling of the size of continuous graphic movements of parkinsonian patients and elderly controls. AB - OBJECTIVES: To systematically investigate the ability of Parkinson's disease patients to discretely and dynamically scale the size of continuous movements and to assess the impact of movement size on outcome variability. METHODS: Ten patients with Parkinson's disease (mean age 72 years) were compared with 12 healthy elderly controls (mean age 70 years). The subjects wrote with a stylus on a graphics tablet. In experiment 1 they drew circles, matching the size of five target circles ranging in magnitude from a radius of 0.5 cm up to 2.5 cm. In experiment 2 they drew spirals with a radius of at least 2 cm. In both experiments the drawings were initially performed as accurately as possible then as fast and accurately as possible. RESULTS: In both experiments the patients and controls drew at a similar speed. The within trial variability of the pen trajectory was greater for patients than controls, and increased disproportionately with the size of the movement. When the emphasis was on size rather than variability (circles), the patients' drawing movements were the same size as controls. When the emphasis was on accuracy of pen trajectory (that is, minimum variability) rather than size (spirals), the patients' drawing movements were smaller than controls. CONCLUSIONS: The movements made by Parkinson's disease patients are hypometric partly as an adaptive strategy used to reduce movement variability. This strategy is used primarily when the requirement to make accurate movements outweighs the need to make large movements. PMID- 12588911 TI - [(123)I]beta-CIT SPECT is a useful method for monitoring dopaminergic degeneration in early stage Parkinson's disease. AB - OBJECTIVES: To examine the validity of [(123)I]beta-CIT SPECT for monitoring the progression of dopaminergic degeneration in Parkinson's disease; to investigate the influence of short term treatment with D(2)receptor agonists on striatal [(123)I]beta-CIT binding; and to determine the sample size and frequency of SPECT imaging required to demonstrate a significant effect of a putative neuroprotective agent. METHODS: A group of 50 early stage Parkinson's disease patients was examined. Two SPECT imaging series were obtained, 12 months apart. The mean annual change in the ratio of specific to non-specific [(123)I]beta-CIT binding to the striatum, putamen, and caudate nucleus was used as the outcome measure. RESULTS: A decrease in [(123)I]beta-CIT binding ratios between the two images was found in all regions of interest. The average decrease in [(123)I]beta CIT binding ratios was about 8% in the whole striatum, 8% in the putaminal region, and 4% in the caudate region. Comparison of scans done in nine patients under two different conditions-in the off state and while on drug treatment showed no significant alterations in the expression of striatal dopamine transporters as measured using [(123)I]beta-CIT SPECT. Power analysis indicated that to detect a significant (p < 0.05) effect of a neuroprotective agent with 0.80 power and 30% of predicted protection within two years, 216 patients are required in each group when the effects are measured in the whole putamen. CONCLUSIONS: [(123)I]beta-CIT SPECT seems to be a useful tool to investigate the progression of dopaminergic degeneration in Parkinson's disease and may provide an objective method of measuring the effectiveness of neuroprotective treatments. Short term treatment with a D(2)agonist does not have a significant influence on [(123)I]beta-CIT binding to dopamine transporters. If the latter finding is replicated in larger groups of patients, it supports the suitability of [(123)I]beta-CIT SPECT for examining the progression of neurodegeneration in patients being treated with D(2)receptor agonists. PMID- 12588913 TI - Neuropsychological and quality of life outcomes 12 months after unilateral thalamic stimulation for essential tremor. AB - OBJECTIVES: To evaluate the one year cognitive, mood state, and quality of life (QoL) outcomes of unilateral thalamic deep brain stimulation (DBS) for essential tremor (ET). METHODS: 40 patients diagnosed with ET completed comprehensive neuropsychological assessments about one month before and three and 12 months after DBS electrode implantation. Data were subjected to multivariate analyses, and significant results were further analysed using univariate techniques. RESULTS: Analyses revealed statistically significant improvements on a cognitive screening measure and in aspects of fine visuomotor and visuoperceptual functions, verbal memory, mood state, and QoL. No group-wise declines in cognition were observed, but more patients showed declines than improvements on language and visual memory tests. Semantic verbal fluency declined significantly in four (10%) of the patients. In these four patients, diminished lexical verbal fluency was present at baseline. CONCLUSION: Cognitive, mood, and QoL outcomes after one year of DBS for ET are favourable; there were no overall deleterious effects on cognition, and DBS was accompanied by a significant reduction in anxiety and improvements in quality of life. However, preoperative verbal fluency diminution may predispose to further fluency declines after DBS. PMID- 12588914 TI - Raised plasma oxidised LDL in acute cerebral infarction. AB - BACKGROUND: The association between oxidised low density lipoprotein (OxLDL) and cerebral infarction is suspected but not established. OBJECTIVES: To determine whether plasma OxLDL is a useful marker for monitoring oxidative stress in stroke patients. METHODS: Plasma OxLDL concentrations were determined in 56 stroke patients with cerebral infarction (n = 45) or cerebral haemorrhage (n = 11), and in 19 age matched controls, using a novel sandwich enzyme linked immunosorbent assay. RESULTS: Compared with the controls (0.130 (0.007) ng/ micro g LDL, mean (SEM)), OxLDL was significantly raised in patients with cerebral infarction (0.245 (0.022); p < 0.0001) but not in those with haemorrhage (0.179 (0.023)). Patients with cortical ischaemic infarcts (n = 22) had higher OxLDL levels than either the controls (p < 0.0001) or the patients with non-cortical ischaemic infarcts (n = 23) (p < 0.001). Increased OxLDL concentrations in patients with cortical infarcts persisted until the third day after stroke onset. The National Institutes of Health stroke scales in patients with cortical infarction were higher than in those with non-cortical infarction (p < 0.01). CONCLUSIONS: There is a significant association between raised plasma OxLDL and acute cerebral infarction, especially cortical infarction. Plasma OxLDL may reflect oxidative stress in stroke patients. PMID- 12588915 TI - Incidence, risk, and case fatality of first ever stroke in the elderly population. The Rotterdam Study. AB - OBJECTIVE: To estimate the incidence, survival, and lifetime risk of stroke in the elderly population. METHODS: The authors conducted a study in 7,721 participants from the population based Rotterdam Study who were free from stroke at baseline (1990-1993) and were followed up for stroke until 1 January 1999. Age and sex specific incidence, case fatality rates, and lifetime risks of stroke were calculated. RESULTS: Mean follow up was 6.0 years and 432 strokes occurred. The incidence rate of stroke per 1,000 person years increased with age and ranged from 1.7 (95% CI 0.4 to 6.6) in men aged 55 to 59 years to 69.8 (95% CI 22.5 to 216.6) in men aged 95 years or over. Corresponding figures for women were 1.2 (95% CI 0.3 to 4.7) and 33.1 (95% CI 17.8 to 61.6). Men and women had similar absolute lifetime risks of stroke (21% for those aged 55 years). The survival after stroke did not differ according to sex. CONCLUSIONS: Stroke incidence increases with age, also in the very old. Although the incidence rate is higher in men than in women over the entire age range, the lifetime risks were similar for both sexes. PMID- 12588916 TI - S100B as a surrogate marker for successful clot lysis in hyperacute middle cerebral artery occlusion. AB - OBJECTIVES: The astroglial protein S100B is a marker of cerebral tissue damage. This study investigated whether the serum kinetic of S100B may serve as a surrogate marker of successful clot lysis and early recanalisation (<6 hours) in hyperacute proximal middle cerebral artery (MCA/M1) occlusion. METHODS: The authors prospectively included 23 patients (mean (SD) age, 70.2 (11.0) years) presenting with MCA/M1 occlusion on magnetic resonance angiography (n=18), intra arterial angiography (IA; n=2), or transcranial Doppler sonography (TCD; n=3) within five hours after symptom onset. Rates of recanalisation and their point of time were determined using TCD or IA. Individual S100B values were determined at hospital admission, every eight hours within the first three days, and at 12 hour intervals from day 4 to day 6. Additionally, the S100B area under the curve (AUC) and the S100B peak value were obtained. RESULTS: Early recanalisation (<6 hours after symptom onset, n=7) was associated with a significantly lower mean S100B AUC compared with no recanalisation (22.2 (40.1) versus 406.8 (284.4) micro g/l per hour; p<0.001). Using receiver operating calculations, a single S100B value obtained 48-96 hours after stroke onset of less than 0.4 micro g/l (cut off point) provided a 86% sensitivity and 100% specificity for sufficient MCA/M1 clot lysis <6 hours. The overall accuracy for a single S100B value obtained in the 48 96 hours time window was as high as for the AUC (95.7%). CONCLUSION: A single S100B value <0.4 micro g/l obtained 48-96 hours after stroke onset indicates successful clot lysis <6 hours in MCA/M1 occlusion with a high degree of accuracy. Thus, determination of a single S100B value may serve as a surrogate marker of early and sufficient MCA/M1 recanalisation in large scale thrombolytic studies. PMID- 12588917 TI - A study of persistent post-concussion symptoms in mild head trauma using positron emission tomography. AB - BACKGROUND: Complaints of persistent cognitive deficits following mild head trauma are often uncorroborated by structural brain imaging and neuropsychological examination. OBJECTIVE: To investigate, using positron emission tomography (PET), the in vivo changes in regional cerebral uptake of 2 [(18)F]fluoro-2-deoxy-D-glucose (FDG) and regional cerebral blood flow (rCBF) in patients with persistent symptoms following mild head trauma. METHODS: Five patients with mild head trauma and five age and education matched healthy controls were imaged using FDG-PET to measure differences in resting regional cerebral glucose metabolism. Oxygen-15 labelled water (H(2)(15)O)-PET was also used to measure group differences in rCBF changes during a spatial working memory task. In addition, neuropsychological testing and self report of dysexecutive function and post-concussion symptoms were acquired to characterise the sample. RESULTS: There was no difference between patients and controls in normalised regional cerebral FDG uptake in the resting state in frontal and temporal regions selected a priori. However, during the spatial working memory task, patients had a smaller increase in rCBF than controls in the right prefrontal cortex. CONCLUSIONS: Persistent post-concussive symptoms may not be associated with resting state hypometabolism. A cognitive challenge may be necessary to detect cerebral changes associated with mild head trauma. PMID- 12588918 TI - Second harmonic imaging: a new ultrasound technique to assess human brain tumour perfusion. AB - BACKGROUND: Second harmonic imaging is a new ultrasound technique that allows evaluation of brain tissue perfusion after application of an ultrasound contrast agent. OBJECTIVE: To evaluate the potential of this technique for the assessment of abnormal echo contrast characteristics of different brain tumours. METHODS: 27 patients with brain tumours were studied. These were divided into four groups: gliomas, WHO grade III-IV (n = 6); meningiomas (n = 9); metastases (n = 5); and others (n = 7). Patients were examined by second harmonic imaging in a transverse axial insonation plane using the transtemporal approach. Following intravenous administration of 4 g (400 mg/ml) of a galactose based echo contrast agent, 62 time triggered images (one image per 2.5 seconds) were recorded and analysed off line. Time-intensity curves of two regions of interest (tumour tissue and healthy brain tissue), including peak intensity (PI) (dB), time to peak intensity (TP) (s), and positive gradient (PG) (dB/s), as well as ratios of the peak intensities of the two regions of interest, were derived from the data and compared intraindividually and interindividually. RESULTS: After administration of the contrast agent a marked enhancement of echo contrast was visible in the tumour tissue in all patients. Mean PI and PG were significantly higher in tumour tissue than in healthy brain parenchyma (11.8 v 5.1 dB and 0.69 v 0.16 dB/s; p < 0.001). TP did not differ significantly (37.1 v 50.2 s; p = 0.14). A tendency towards higher PI and PG as well as shorter TP was apparent in malignant gliomas. When comparing different tumour types, however, none of these variables reached significance, nor were there significant differences between malignant and benign tumours in general. CONCLUSIONS: Second harmonic imaging not only allows identification of brain tumours, but may also help in distinguishing between different tumour types. It gives additional and alternative information about tumour perfusion. Further studies are needed to evaluate the clinical potential of this technique in investigating brain tumours-for example in follow up investigations of patients undergoing radiation or chemotherapy-especially in comparison with neuroradiological and neuropathological findings. PMID- 12588919 TI - Muscle hypertrophy in myotonia congenita. PMID- 12588920 TI - A controlled study comparing visual function in patients treated with vigabatrin and tiagabine. AB - OBJECTIVE: Vigabatrin treatment is frequently associated with irreversible retinal injury and produces retinal electrophysiological changes in nearly all patients. Concern has been raised that tiagabine and other antiepilepsy drugs (AEDs) that increase brain gamma-aminobutyric acid (GABA) might produce similar electrophysiological and clinical changes in visual function. The study compared visual function between groups of patients with epilepsy treated long term with tiagabine, vigabatrin, and patients treated with other AEDs. METHODS: A cross sectional study comparing visual acuity, colour vision, static and kinetic perimetry, and electroretinograms between groups of patients treated with tiagabine, vigabatrin, and other AEDs (control patients). Patients were adults receiving stable AED treatment for >6 months. RESULTS: Vigabatrin treated patients had marked visual field constrictions in kinetic perimetry (mean radius 39.6 degrees OD, 40.5 degrees OS), while tiagabine patients had normal findings (mean 61 degrees OD, 62 degrees OS) (differences OD and OS, p=0.001), which were similar to epilepsy control patients (mean 60 degrees OD, 61 degrees OS). Vigabatrin patients had abnormal electroretinographic photopic B wave, oscillatory, and flicker responses, which correlated with visual field constrictions. These electroretinographic responses were normal for tiagabine patients and control patients. Patients were treated with vigabatrin for a median of 46 months compared with 29 months for tiagabine. Patients taking other AEDs that may change brain GABA had normal visual function. CONCLUSION: Unlike vigabatrin, tiagabine treatment is associated with normal electroretinography and visual fields and ophthalmological function similar to epilepsy control patients. Differences between vigabatrin and other GABA modulating AEDs in retinal drug concentrations and other effects might explain why tiagabine increases in GABA reuptake do not cause retinal injury. PMID- 12588921 TI - Allvar Gullstrand (1862-1930). PMID- 12588922 TI - Hypothesis on the pathophysiology of syringomyelia based on simulation of cerebrospinal fluid dynamics. AB - OBJECTIVES: Despite many hypotheses, the pathophysiology of syringomyelia is still not well understood. In this report, the authors propose a hypothesis based on analysis of cerebrospinal fluid dynamics in the spine. METHODS: An electric circuit model of the CSF dynamics of the spine was constructed based on a technique of computational fluid mechanics. With this model, the authors calculated how a pulsatile CSF wave coming from the cranial side is propagated along the spinal cord. RESULTS: Reducing the temporary fluid storage capacity of the cisterna magna dramatically increased the pressure wave propagated along the central canal. The peak of this pressure wave resided in the mid-portion of the spinal cord. CONCLUSIONS: The following hypotheses are proposed. The cisterna magna functions as a shock absorber against the pulsatile CSF waves coming from the cranial side. The loss of shock absorbing capacity of the cisterna magna and subsequent increase of central canal wall pressure leads to syrinx formation in patients with Chiari I malformation. PMID- 12588923 TI - Agreement among neurologists on the clinical diagnosis of dystonia at different body sites. AB - OBJECTIVE: To study the reliability of the diagnosis of blepharospasm, oromandibular dystonia, cervical dystonia, and writer's cramp among neurologists. METHODS: 12 patients with adult onset focal segmental dystonia were videotaped in a standardised way. The tape was sent to six neurologists who are involved in clinical practice without a specific interest in movement disorders (general neurologists), and to four neurologists expert in movement disorders. The observers had to recognise whether the patients were affected by dystonia and to distinguish among blepharospasm, oromandibular dystonia, cervical dystonia, and writer's cramp. Interobserver reliability was assessed by kappa statistics, and the degree of agreement was classified according to the Landis classification. RESULTS: The 10 neurologists reached slight to moderate agreement on the diagnosis of these four disorders. When the observers were subdivided according with their professional experience in the field, a moderate to perfect agreement on the diagnosis was achieved by specialists in movement disorders, and a fair to moderate agreement by the general neurologists. CONCLUSIONS: Neurologists may have different ability to recognise adult onset focal dystonia, depending on their experience and on the type of dystonia. PMID- 12588924 TI - Diagnostic criteria and the use of ICD-10 codes to define and classify minor head injury. AB - BACKGROUND: Epidemiological research on the incidence of traumatic head injuries relies on the correct definition and classification of the injury. OBJECTIVE: To address the use of diagnostic criteria and ICD-10 codes to define minor head injury in Swedish hospitals managing patients with head injury. METHODS: A questionnaire was mailed to all 76 Swedish hospitals managing head injuries. The hospitals were asked what diagnostic criteria they use to define minor head injury, and which ICD-10 codes they use to classify such injuries. RESULTS: 72 hospitals (95%) responded to the survey. The most common criterion was loss of consciousness (76%), followed by post-traumatic amnesia (38%). Almost half the hospitals used other signs and symptoms to define minor head injury. The ICD-10 code S.06 (intracranial injury) was used by 51 of the hospitals (91%). CONCLUSIONS: It is essential that there should be common definitions, classifications, and registration of minor head injuries. The wide variation in definition and classification found in this study emphasises the importance of improved implementation of the present guidelines. PMID- 12588925 TI - Central pontine myelinolysis associated with hypokalaemia in anorexia nervosa. AB - A 31-year-old man was admitted to hospital with of anorexia, binge eating, and self induced vomiting. On admission, he showed a pronounced low weight and disturbance of the body image and was diagnosed as having anorexia nervosa. In addition, electrolyte abnormalities, mainly hypokalaemia, and increased serum renin and aldosterone concentrations were recorded, suggesting pseudo-Bartter syndrome as a complication. Under frequent monitoring of the serum potassium and sodium concentrations, serum electrolytes were gradually corrected, but brain magnetic resonance imaging revealed reversible central pontine myelinolysis (CPM). Although attention has been mainly paid to the association of CPM with rapid correction of hyponatraemia and abnormal osmolality, this case report strongly suggested the involvement of hypokalaemia in the pathogenesis of CPM. PMID- 12588926 TI - Eyelid tremor in a patient with a unilateral paramedian thalamic lesion. AB - A patient with a circumscribed infarction of the right paramedian thalamus developed a tremor of both eyelids on voluntary eye closure. Co-registration of the magnetic resonance image to a stereotactic atlas of the human thalamus revealed that the lesion was confined to a small subgroup of paramedian nuclei, including the parvocellular part of the mediodorsal nucleus. It is concluded that this region provides inhibitory input to cortical and/or subcortical regions controlling eyelid movements. Voluntary eye closure may involve direct cortico nuclear connections and indirect pathways through the paramedian thalamus, most probably through the mediodorsal nucleus. PMID- 12588927 TI - Is neurocysticercosis a risk factor in coexistent intracranial disease? An MRI based study. AB - BACKGROUND: Previous reports have suggested that neurocysticercosis is associated with glioma and Japanese encephalitis, and that it is a risk factor for stroke. OBJECTIVE: To determine if neurocysticercosis has a significant association with, or is a risk factor for, coexistent pathologies such as Japanese encephalitis, glioma, abscess, tuberculoma, or infarction. SUBJECTS: 10 350 patients from the hospital population who underwent 1.5 T cranial magnetic resonance imaging during the previous 12 years were evaluated for the presence of neurocysticercosis and coexisting pathology. DESIGN: Retrospective cohort analysis. RESULTS: The prevalence of neurocysticercosis in cases with dual pathology was significantly less than in a control group (1.1% v 8.3%; z = 11.05; p < 0.001, power of test = 1). Neurocysticercosis lesions were less common (p < 0.05) in the different subgroups of coexistent pathology than in the control group except in the case of Japanese encephalitis, where the difference was non-significant (z = 0.69, p = 0.49). The relative risk was less than 1 in all subgroups except Japanese encephalitis, where it was 1.23. The location of neurocysticercosis lesions and the presence of perilesional oedema did not affect coexistent lesion location or severity on a particular side (p = 0.413 and 0.623 for location and perilesional oedema, respectively). When the above factors were analysed separately in patients with Japanese encephalitis, they also did not affect coexistent lesion location or severity (p = 0.659 and 0.548, respectively). CONCLUSIONS: The coexistence of neurocysticercosis and other lesions may be an incidental observation in a few patients referred from areas of high prevalence and endemicity. It appears unlikely that neurocysticercosis is a risk factor for other intracerebral pathology. The location of neurocysticercosis lesions and whether or not there is surrounding perilesional oedema do not appear to affect the location or severity of coexisting lesions. PMID- 12588928 TI - Saitohin gene is not associated with Alzheimer's disease. AB - BACKGROUND: The deposition of tau protein in neurofibrillary tangles constitutes an important feature of many neurodegenerative disorders, including Alzheimer's disease. A polymorphic gene, saitohin (STH), nested within the tau gene (microtubule associated protein tau, MAPT), was recently identified and an association of a non-synonymous polymorphism in STH with increased risk for Alzheimer's disease was suggested. OBJECTIVE AND METHODS: To test the above hypothesis in a case-control association study of two independent white populations within Switzerland and Greece, comparing genotype and allele frequencies from 225 Alzheimer's disease patients and 144 healthy control subjects. RESULTS: No differences in allelic or genotypic distributions between Alzheimer's disease patients and controls was found in the individual samples (Swiss/Greek) or in the combined sample. Stratification for the presence of apolipoprotein E (APOE) epsilon 4 allele, sex, or age did not show significant effects in the populations studied, nor was there an effect on the age of onset. CONCLUSIONS: No evidence was found for an association of the non-synonymous polymorphism (Q7R) in STH and Alzheimer's disease. This finding is in line with earlier studies showing no association between MAPT and Alzheimer's disease. PMID- 12588929 TI - A case of gliosarcoma appearing as ischaemic stroke. AB - OBJECTIVES: Ischaemic stroke attributable to malignant brain tumour is a rarely reported phenomenon and even various imaging techniques including angiography do not necessarily lead to an accurate diagnosis. CASE DESCRIPTION: A 46-year-old, previously healthy man developed apoplectic symptoms with slight right sided hemiparesis and global aphasia. The computed tomography (CT) scan showed lesions of the left temporal lobe and the paraventricular white matter suggestive of left middle cerebral artery (MCA) infarction. Carotid angiography demonstrated compression of the M1 segment of the MCA and occlusion of temporal MCA. The patient initially refused magnetic resonance imaging (MRI) because of claustrophobia. Because of fluctuating symptoms and successive worsening of the condition over weeks an MRI scan was conducted under general anaesthesia. Beneath temporal, opercular, and subcortical infarctions it revealed a left temporal tumour. A tumour biopsy disclosed a gliosarcoma (WHO grade IV). Microscopical examination of the surgical specimen demonstrated invasion of tumour cells into the wall of a greater pre-existing blood vessel. CONCLUSIONS: Malignant brain tumours may cause ischaemic infarction. This is a rare but important differential diagnosis for the origin of strokes. The authors describe the first case with infiltration of intracranial blood vessels by tumour cells of a gliosarcoma. PMID- 12588931 TI - Endplate dysfunction causing respiratory failure in a patient with prior paralytic poliomyelitis. AB - A 56-year-old man with late amyotrophic sequelae from poliomyelitis experienced progressive dyspnoea requiring intubation and artificial ventilation in the intensive care unit. Repetitive stimulation studies showed a marked decrement of the trapezius muscle response reversible with edrophonium. Ventilatory function considerably and lastingly improved under anticholinesterase treatment. In the absence of biological evidence for autoimmune myasthenia gravis, it is suggested that a mechanism implying endplate dysfunction related to postpolio syndrome. Repetitive stimulation procedure should be considered in postpolio syndrome patients as some of them may benefit from anticholinesterase treatment. PMID- 12588930 TI - Seizures arising from the inferior parietal lobule can show ictal semiology of the second sensory seizure (SII seizure). AB - A 52-year-old right handed man presented with medically intractable partial seizures consisting of numbness on the left upper back spreading to the left upper as well as lower limbs. Head computed tomography and magnetic resonance imaging showed a round calcified lesion in the depth of the superior ramus of the right sylvian fissure. Ictal electrocorticographic recording with chronically implanted subdural electrodes showed low voltage fast activities starting exclusively from an electrode located on the right inferior parietal lobule. No apparent ictal activities were observed from the depth electrodes inserted in the parietal operculum. Somatosensory evoked potentials of 75 ms to 145 ms latency were recorded from the ictal onset zone, which was 2 cm caudal to the perisylvian area corresponding to the second somatosensory area. Seizures arising from the inferior parietal lobule including the angular and supuramarginal gyri can produce partial seizures whose ictal semiology and scalp electroencephalography are indistinguishable from the ones originating from the second somatosensory area. PMID- 12588932 TI - Improvement of central motor conduction after bone marrow transplantation in adrenoleukodystrophy. AB - The case is described of a 20-year-old man with adrenoleukodystrophy who showed right spastic hemiparesis and gait disturbance. Brain magnetic resonance imaging disclosed predominant involvement of the left corticospinal pathway. The clinical symptoms improved after bone marrow transplantation. Transcranial magnetic stimulation disclosed significant improvement in various parameters of central motor conduction. PMID- 12588933 TI - Decision making in humans: the effect of manipulating the central noradrenergic system. AB - OBJECTIVES: Damasio has proposed an influential model of human decision making the Somatic Marker Hypothesis (SMH), where he argues that somatic feedback to the brain influences decision making in humans. It is proposed that when choosing between options that differ in relative risk, a somatic marker (for example, a "gut feeling") feeds back to the brain and influences cognitive appraisal. This study tested the hypothesis that the noradrenergic system may play a part in this afferent feedback process. METHODS: Thirty healthy young adults were randomly allocated to receive placebo, 4 mg, or 8 mg reboxetine (a new selective noradrenergic reuptake inhibitor that acts to increase central noradrenergic activity). It is hypothesised that enhancement of central noradrenergic activity may amplify the effect of afferent feedback, and lead to altered decision making. All subjects completed the Iowa Gambling Task (IGT), a computerised card playing game where the player is instructed to try and win as much money as possible over 100 selections from one of four decks. RESULTS: There were no significant differences between the three groups in gambling task performance. CONCLUSION: This result suggests that central noradrenergic activity may not be critically involved in the afferent feedback mechanism that is central to the somatic marker hypothesis. PMID- 12588934 TI - Repetitive stimulation of the long thoracic nerve in myasthenia gravis: clinical and electrophysiological correlations. AB - OBJECTIVE: To investigate repetitive stimulation of the long thoracic nerve as a reliable and technically undemanding method for evaluating respiratory muscle function in myasthenia gravis. METHODS: 15 patients with myasthenia gravis and positive single fibre electromyography had repetitive stimulation of the long thoracic nerve together with serratus anterior recordings. There were 20 normal controls. RESULTS: Eight patients with respiratory symptoms all had abnormal results, with an increased response decrement. These patients required intensive monitoring and supplemental oxygen. The remaining seven cases with no respiratory symptoms all had negative studies. CONCLUSIONS: This is a simple well tolerated method that shows good correlation with respiratory symptoms and management requirements in myasthenia gravis. It can alert the physician to serious respiratory complications in this disease. PMID- 12588935 TI - Diffusion weighted imaging in radiation necrosis. AB - This case report suggests that magnetic resonance imaging with diffusion weighted imaging may help distinguish between tumour recurrence and radiation induced necrosis in patients previously treated for a brain tumour. PMID- 12588936 TI - Neurofibromatosis. PMID- 12588937 TI - Neuroleptic malignant syndrome in Kufs' disease. AB - A patient with adult neuronal ceroid lipofuscinosis (ANCL; Kufs' disease) is described in whom neuroleptic malignant syndrome occurred, initially presenting as catatonic syndrome. Comprehensive neuroimaging studies were conducted including FDG-PET, IBZM-SPECT, and beta-CIT-SPECT, electrophysiological examinations and an ex vivo contracture test exposing muscle biopsy specimens to neuroleptics. Collectively the results argued for an involvement of the muscle in neuroleptic malignant syndrome at least in ANCL. PMID- 12588938 TI - Acute rotatory vertigo caused by a small haemorrhage of the vestibular cortex. PMID- 12588939 TI - Reversible callosal disconnection syndrome in internal hydrocephalus. PMID- 12588940 TI - Coexistent Lewy body disease in a case of "visual variant of Alzheimer's disease". PMID- 12588941 TI - Trigeminal autonomic cephalalgia-tic-like syndrome associated with a pontine tumour in a one-year-old girl. PMID- 12588942 TI - Autoimmune myasthenia gravis after cardiac surgery. PMID- 12588943 TI - Expanding cerebral cysts (lacunae): a treatable cause of progressive midbrain syndrome. PMID- 12588944 TI - Chiari I malformation mimicking myasthenia gravis. PMID- 12588945 TI - Conflict and integration of spatial attention between disconnected hemispheres. PMID- 12588946 TI - Measurement of intestinal cholesterol absorption by plasma and fecal dual-isotope ratio, mass balance, and lymph fistula methods in the mouse: an analysis of direct versus indirect methodologies. AB - The rate of intestinal cholesterol (Ch) absorption is an important criterion for quantitation of Ch homeostasis. However, studies in the literature suggest that percent Ch absorption, measured usually by a fecal dual-isotope ratio method, spans a wide range, from 20% to 90%, in healthy inbred mice on a chow diet. In the present study, we adapted four standard methods, one direct (lymph collection) and three indirect (plasma and fecal dual-isotope ratio, and sterol balance) measurements of Ch absorption and applied them to mice. Our data establish that all methodologies can be valid in mice, with all methods supporting the concept that gallstone-susceptible C57L mice absorb significantly more Ch (37 +/- 5%) than gallstone-resistant AKR mice (24 +/- 4%). We ascertained that sources of error in the literature leading to marked differences in Ch absorption efficiencies between laboratories relate to a number of technical factors, most notably expertise in mouse surgery, complete solubilization and delivery of radioisotopes, appropriate collection periods for plasma and fecal samples, and total extraction of radioisotopes from feces. We find that all methods provide excellent interexperimental agreement, and the ranges obtained challenge previously held beliefs regarding the spread of intestinal Ch absorption efficiencies in mice. The approaches documented herein provide quantifiable methodologies for exploring genetic mechanisms of Ch absorption, and for investigating the assembly and secretion of chylomicrons, as well as intestinal lipoprotein metabolism in mice. PMID- 12588947 TI - A procedure for preparing oxazolines of highly unsaturated fatty acids to determine double bond positions by mass spectrometry. AB - A convenient, mild, reliable method has been developed for preparing oxazolines of fatty acids and for using these derivatives to determine double bond locations in long-chain polyunsaturated and polyconjugated fatty acids. Fatty acyl mixed anhydrides are prepared using isobutylchloroformate and then converted to their ethanolamides by treatment with ethanolamine. Ethanolamides are subsequently cyclized to the corresponding oxazolines in >or=85% yields by treatment with trifluoroacetic anhydride under mild conditions (>50 degrees for 30-60 min). This general protocol can also be used to synthesize 4,4-dimethyloxazoline and benzoxazole derivatives of fatty acids. Gas chromatography-mass spectrometry of oxazoline derivatives of fatty acids yields prominent ions diagnostic of the structures of the parent fatty acids and, in the case of unsaturated fatty acids, indicating the positions of the double bonds. The utility of the method is illustrated with several fatty acids, including the conjugated 4E,6E,8E,10E,13Z,16Z,19Z-docosaheptaenoic acid. PMID- 12588948 TI - A CYP7A promoter binding factor site and Alu repeat in the distal promoter region are implicated in regulation of human CETP gene expression. AB - The cholesteryl ester transfer protein (CETP) plays a key role in reverse cholesterol transport in mediating the transfer of cholesteryl ester from HDL to atherogenic apolipoprotein B-containing lipoproteins (VLDL, IDL, and LDL). Variation in plasma CETP mass in both normolipidemic and dyslipidemic individuals may reflect differences in CETP gene expression. As the 5' flanking sequence up to 3.4 kb of the human CETP gene contributes to transcriptional activity and tissue-specific gene expression, we evaluated the role of the distal promoter region in the modulation of CETP gene expression. In transfection experiments in HepG2 cells, we presently demonstrate that an Alu repeat (-2,153/-2,414) acts as a repressive element, whereas a binding site for the orphan nuclear receptor CYP7A promoter binding factor (CPF), at position -1,042, facilitates activation of human CETP promoter activity. Cotransfection of liver receptor homolog, the mouse homologue of CPF in HEK293 cells that lack CPF, indicated that the -1,042 CPF site is sufficient to induce CPF-mediated activation of CETP promoter activity. Taken together, our results indicate that the distal-promoter region is a major component in the modulation of human CETP promoter activity, and that it may contribute to the liver-specific expression of the CETP gene. PMID- 12588949 TI - Covalent binding of hydroxy-alkenals 4-HDDE, 4-HHE, and 4-HNE to ethanolamine phospholipid subclasses. AB - Lipid oxidation is implicated in a wide range of pathophysiogical disorders, and leads to reactive compounds such as fatty aldehydes, of which the most well known is 4-hydroxy-2E-nonenal (4-HNE) issued from 15-hydroperoxyeicosatetraenoic acid (15-HpETE), an arachidonic acid (AA) product. In addition to 15-HpETE, 12(S) HpETE is synthesized by 12-lipoxygenation of platelet AA. We first show that 12 HpETE can be degraded in vitro into 4-hydroxydodeca-(2E,6Z)-dienal (4-HDDE), a specific aldehyde homologous to 4-HNE. Moreover, 4-HDDE can be detected in human plasma. Second, we compare the ability of 4-HNE, 4-HDDE, and 4-hydroxy-2E-hexenal (4-HHE) from n-3 fatty acids to covalently modify different ethanolamine phospholipids (PEs) chosen for their biological relevance, namely AA- (20: 4n-6) or docosahexaenoic acid- (22:6n-3) containing diacyl-glycerophosphoethanolamine (diacyl-GPE) and alkenylacyl-glycerophosphoethanolamine (alkenylacyl-GPE) molecular species. The most hydrophobic aldehyde used, 4-HDDE, generates more adducts with the PE subclasses than does 4-HNE, which itself appears more reactive than 4-HHE. Moreover, the aldehydes show higher reactivity toward alkenylacyl-GPE compared with diacyl-GPE, because the docosahexaenoyl-containing species are more reactive than those containing arachidonoyl. We conclude that the different PE species are differently targeted by fatty aldehydes: the higher their hydrophobicity, the higher the amount of adducts made. In addition to their antioxidant potential, alkenylacyl-GPEs may efficiently scavenge fatty aldehydes. PMID- 12588950 TI - Hypercholesterolemia and changes in lipid and bile acid metabolism in male and female cyp7A1-deficient mice. AB - Cholesterol 7alpha-hydroxylase, a rate-limiting enzyme for bile acid synthesis, has been implicated in genetic susceptibility to atherosclerosis. The gene, CYP7A1, encoding a protein with this activity, is expressed normally only in hepatocytes and is highly regulated. Our cyp7A1 gene knockout mouse colony, as young adults on a chow diet, is hypercholesterolemic. These mice were characterized extensively to understand how cyp7A1 affects lipid and bile acid homeostasis in different tissue compartments and whether gender plays a modifying role. Both male and female cyp7A1-deficient mice had decreased hepatic LDL receptors, unchanged hepatic cholesterol synthesis, increased intestinal cholesterol synthesis and bile acid transporters, and decreased fecal bile acids but increased fecal sterols. In females, cyp7A1 deficiency also caused changes in hepatic fatty acid metabolism, decreased hepatic canalicular bile acid transporter, Bsep, and gallbladder bile composition altered to a lithogenic profile. Taken together, the data suggest that cyp7A1 deficiency results in a proatherogenic phenotype in both genders and leads to a prolithogenic phenotype in females. PMID- 12588951 TI - Quantitative trait loci that determine lipoprotein cholesterol levels in DBA/2J and CAST/Ei inbred mice. AB - To investigate genetic contributions to individual variations of lipoprotein cholesterol concentrations, we performed quantitative trait locus/loci (QTL) analyses of an intercross of CAST/Ei and DBA/2J inbred mouse strains after feeding a high-cholesterol cholic acid diet for 10 weeks. In total, we identified four QTL for HDL cholesterol. Three of these were novel and were named Hdlq10 [20 centimorgans (cM), chromosome 4], Hdlq11 (48 cM, chromosome 6), and Hdlq12 (68 cM, chromosome 6). The fourth QTL, Hdl1 (48 cM, chromosome 2), confirmed a locus discovered previously using a breeding cross that employed different inbred mouse strains. In addition, we identified one novel QTL for total and non-HDL cholesterol (8 cM, chromosome 9) that we named Chol6. Hdlq10, colocalized with a mutagenesis-induced point mutation (Lch), also affecting HDL. We provide molecular evidence for Abca1 as the gene underlying Hdlq10 and Ldlr as the gene underlying Chol6 that, coupled with evidence generated by other researchers using knockout and transgenic models, causes us to postulate that polymorphisms of these genes, different from the mutations leading to Tangier's disease and familial hypercholesterolemia, respectively, are likely primary genetic determinants of quantitative variation of lipoprotein levels in mice and, by orthology, in the human population. PMID- 12588952 TI - Blocking microsomal triglyceride transfer protein interferes with apoB secretion without causing retention or stress in the ER. AB - Microsomal triglyceride transfer protein (MTP) is an intraluminal protein in the endoplasmic reticulum (ER) that is essential for the assembly of apolipoprotein B (apoB)-containing lipoproteins. In this study, we examine how the livers of mice respond to two distinct methods of blocking MTP function: Cre-mediated disruption of the gene for MTP and chemical inhibition of MTP activity. Blocking MTP significantly reduced plasma levels of triglycerides, cholesterol, and apoB containing lipoproteins in both wild-type C57BL/6 and LDL receptor-deficient mice. While treating LDL receptor-deficient mice with an MTP inhibitor for 7 days lowered plasma lipids to control levels, liver triglyceride levels were increased by only 4-fold. Plasma levels of apoB-100 and apoB-48 fell by >90% and 65%, respectively, but neither apoB isoform accumulated in hepatic microsomes. Surprisingly, loss of MTP expression was associated with a nearly complete absence of apoB-100 in hepatic microsomes. Levels of microsomal luminal chaperone proteins [e.g., protein disulfide isomerase, glucose-regulated protein 78 (GRP78), and GRP94] and cytosolic heat shock proteins (HSPs) (e.g., HSP60, HSC, HSP70, and HSP90) were unaffected by MTP inhibition. These findings show that the liver responds rapidly to inhibition of MTP by degrading apoB and preventing its accumulation in the ER. The rapid degradation of secretion-incompetent apoB in the ER may block the induction of proteins associated with unfolded protein and heat shock responses. PMID- 12588953 TI - Effect of apoC-III gene polymorphisms on the lipoprotein-lipid profile of viscerally obese men. AB - Abdominal visceral adipose tissue (AT) accumulation is associated with an atherogenic metabolic profile that includes increased plasma triglyceride (TG), low HDL cholesterol levels, and an insulin-resistant hyperinsulinemic state. Whereas the apolipoprotein (apo) C-III C3238G gene variant, often referred to as the SstI polymorphism, has been related to variations in plasma TG concentrations, another variation within the insulin responsive element (C-482T) of the apoC-III gene has been associated with greater glucose and insulin responses to an oral glucose tolerance test (OGTT); however, these results were obtained in nonobese individuals. We therefore investigated the effects of three apoC-III gene polymorphisms, namely SstI, C-482T, and T-455C, on fasting plasma lipoprotein-lipid levels and response to a 75 g OGTT in a sample of 122 viscerally obese men (abdominal visceral AT area >or=130 cm(2)). Among the three gene variants that were examined, the SstI variation was the only one found to be associated with hypertriglyceridemia. Indeed, S1/S2 heterozygotes (n = 24) were characterized by increased fasting plasma TG concentrations compared with S1/S1 homozygotes (n = 98) (mean +/- SD: 3.03 +/- 1.58 vs. 2.34 +/- 0.95 mmol/l respectively, P < 0.05). The higher TG concentrations in S1/S2 were associated with the presence of smaller, denser LDL particles compared with S1/S1 subjects (LDL peak particle diameter: 24.8 +/- 0.5 nm vs. 25.1 +/- 0.5 nm respectively, P < 0.05). Furthermore, there was no association between the response to the OGTT and any of the apoC-III gene variants (SstI, T-455C, or C-482T) examined. Results of the present study support the notion of a hypertriglyceridemic effect associated with the apoC-III SstI polymorphism that could modulate the magnitude of the dyslipidemic state in abdominally obese patients. PMID- 12588954 TI - Visualization of identified GFP-expressing cells by light and electron microscopy. AB - We have developed a procedure for visualizing GFP expression in fixed tissue after embedding in LR White. We find that GFP fluorescence survives fixation in 4% paraformaldehyde/0.1% glutaraldehyde and can be visualized directly by fluorescence microscopy in unstained, 1 microm sections of LR White-embedded material. The antigenicity of the GFP is retained in these preparations, so that GFP localization can be visualized in the electron microscope after immunogold labeling with anti-GFP antibodies. The ultrastructural morphology of tissue fixed and embedded by this protocol is of quality sufficient for subcellular localization of GFP. Thus, expression of GFP constructs can be visualized in living tissue and the same cells relocated in semithin sections. Furthermore, semithin sections can be used to locate GFP-expressing cells for examination by immunoelectron microscopy of the same material after thin sectioning. PMID- 12588955 TI - Immunocytochemical detection of phosphatidylinositol 3-kinase activation by insulin and leptin. AB - Intracellular signaling mediated by phosphatidylinositol 3-kinase (PI3K) is important for a number of cellular processes and is stimulated by a variety of hormones, including insulin and leptin. A histochemical method for assessment of PI3K signaling would be an important advance in identifying specific cells in histologically complex organs that are regulated by growth factors and peptide hormones. However, current methods for detecting PI3K activity require either homogenization of the tissue or cells or the ability to transfect probes that bind to phosphatidylinositol 3,4,5 trisphosphate (PIP3), the reaction product of PI3K catalysis. Here we report the validation of an immunocytochemical method to detect changes in PI3K activity, using a recently developed monoclonal antibody to PIP3, in paraformaldehyde-fixed bovine aortic endothelial cells (BAECs) in culture and in hepatocytes of intact rat liver. Treatment with either insulin or leptin increased BAEC PIP3 immunoreactivity, and these effects were blocked by pretreatment with PI3K inhibitors. Furthermore, infusion of insulin into the hepatic portal vein of fasted rats caused an increase of PIP3 immunostaining in hepatocytes that was associated with increased serine phosphorylation of the downstream signaling molecule protein kinase B/Akt (PKB/Akt). We conclude that immunocytochemical PIP3 staining can detect changes in PI3K activation induced by insulin and leptin in cell culture and intact liver. PMID- 12588956 TI - The collagen type XVIII endostatin domain is co-localized with perlecan in basement membranes in vivo. AB - The C-terminal globular endostatin domain of collagen type XVIII is anti angiogenic in a variety of experimental tumor models, and clinical trials to test it as an anti-tumor agent are already under way. In contrast, many of its cell biological properties are still unknown. We systematically localized the mRNA of collagen type XVIII with the help of in situ hybridization (ISH) and detected it in epithelial and mesenchymal cells of almost all organ systems throughout mouse development. Light and electron microscopic immunohistochemistry (IHC) revealed that the endostatin domain is a widespread component of almost all epithelial basement membranes in all major developing organs, and in all basement membranes of capillaries and blood vessels. Furthermore, quantitative immunogold double labeling demonstrated a co-localization of 50% of the detected endostatin domain together with perlecan in basement membranes in vivo. We conclude that the endostatin domain of collagen type XVIII plays a role, even in early stages of mouse development, other than regulating angiogenesis. In the adult, the endostatin domain could well be involved in connecting collagen type XVIII to the basement membrane scaffolds. At least in part, perlecan appears to be an adaptor molecule for the endostatin domain in basement membranes in vivo. PMID- 12588957 TI - NAD(P)H: quinone oxidoreductase 1 expression in kidney podocytes. AB - NAD(P)H:quinone oxidoreductase 1 (NQO1; DT-diaphorase; DTD) is a cytosolic two electron reductase, and compounds of the quinone family such as mitomycin C are efficiently bioactivated by this enzyme. The observation that DT-diaphorase is highly expressed in many cancerous tissues compared to normal tissues has provided us with a potentially selective target that can be exploited in the design of novel anticancer agents. Because of the relative lack of information about the cell-specific expression of DT-diaphorase, the purpose of this study was to map the distribution of this enzyme in normal human tissues. Fifteen tissue samples from normal human kidney were analyzed for expression of DT diaphorase by immunohistochemistry (two-step indirect method). We found a specific high expression of DT-diaphorase in glomerular visceral epithelial cells (podocytes). These results suggest that a high expression of DT-diaphorase in podocytes could play a major role in the pathogenesis of renal toxicity and mitomycin C-induced hemolytic uremic syndrome, in which injury to the glomerular filtration mechanism is the primary damage, leading to a cascade of deleterious events including microangiopathic hemolytic anemia and thrombocytopenia. This observation has potential therapeutic implications because the DT-diaphorase metabolic pathway is influenced by many agents, including drugs, diet, and environmental cell factors such as pH and oxygen tension. PMID- 12588958 TI - DNA ploidy and markovian analysis of neoplastic progression in experimental pancreatic cancer. AB - Computer-assisted analysis of DNA ploidy and nuclear morphology were used to elucidate changes in the cell nucleus that occur during the development of experimental pancreatic cancer. Ductal pancreatic adenocarcinoma was induced in 49 Syrian hamsters by SC injection of N-nitrosobis (2-oxopropyl) amine; twenty hamsters served as controls. Groups of animals were sacrificed every 4 weeks for 20 weeks and adjacent sections of pancreatic tissue were H&E and Feulgen-stained for light microscopy and computer assisted cytometry. Pancreatic ductal cells were classified as normal, atypical, or malignant; tissue inflammation (pancreatitis) was also noted when present. DNA ploidy and nuclear morphology evaluation (Markovian analysis) identified an atypical cell stage clearly distinguishable from either normal or malignant cells; pancreatitis preceded this atypia. The DNA ploidy histogram of these atypical cells revealed a major diploid peak and a minor aneuploid peak. The receiver operator characteristic curve areas for a logistic regression model of normal vs atypical cells was 0.94 and for atypical vs malignant was 0.98, numbers indicative of near-perfect discrimination among these three cell types. The ability to identify an atypical cell population should be useful in establishing the role of these cells in the progression of human pancreatic adenocarcinoma. PMID- 12588959 TI - Immunolocalization of aromatase in stallion Leydig cells and seminiferous tubules. AB - High levels of plasma estrogens constitute an endocrine peculiarity of the adult stallion. This is mostly due to testicular cytochrome p450 aromatase, the only irreversible enzyme responsible for the bioconversion of androgens into estrogens. To identify more precisely the testicular aromatase synthesis sites in the stallion, testes from nine horses (2-5 years) were obtained during winter or spring. Paraplast-embedded sections were processed using rabbit anti-equine aromatase, followed by biotinylated goat anti-rabbit antibodies, and amplified with a streptavidin-peroxidase complex. Immunoreactivity was detected with diaminobenzidine. Immunofluorescence detection, using fluoroisothiocyanate conjugated goat anti-rabbit antibodies, was also applied. Specific aromatase immunoreactivity was observed intensely in Leydig cells but also for the first time, to a lesser extent, in the cytoplasm surrounding germ cells at the junction with Sertoli cells. Interestingly, the immunoreactivity in Sertoli cells appears to vary with the spermatogenic stages in the basal compartment (with spermatogonia) as well as in the adluminal one (with spermatids). Relative staining intensity in Leydig and Sertoli cells and testicular microsomal aromatase activity increased with age. The present study in stallions indicates that in addition to Leydig cells, Sertoli cells also appear to participate in estrogen synthesis, and this could play a paracrine role in the regulation of spermatogenesis. PMID- 12588960 TI - Quantitative fluorescence imaging approach for the study of polyploidization in hepatocytes. AB - We applied automatic quantitative fluorescence imaging of nuclear DNA to rat liver cells obtained from animals at various times after birth up to 3 months of age. We show that, in conditions best preserving the native cellular structures, DNA content measurements, performed on whole single cells in situ after Hoechst staining, were precise and accurate. Cells in the various ploidy and nuclearity classes could thus be identified correctly and their percentages were estimated on a total of 300 cells or more. DNA synthesis was shown to occur asynchronously in all ploidy and nuclearity classes around weaning time. Observation of the labeling patterns, after in vivo BrdU pulse and short-term culture (chase), showed that the cell cycle was shorter in diploid cells compared with cells undergoing polyploidization. These results show that the approach of fluorescence imaging is well suited to investigations on polyploidization mechanisms. PMID- 12588961 TI - Lymphatic network and lymphangiogenesis in the gastric wall. AB - A family of growth factors highly specific for endothelial cells was identified more than 10 years ago, in which the receptor of vascular endothelial growth factor C (VEGFR-3) is implicated in the regulation of lymphatic development and regeneration. Comparative studies on the lymphatic network and lymphangiogenesis have been done mainly using combined 5'-nucleotidase (5'-Nase) enzyme and VEGFR-3 immunohistochemical approaches in adult and fetal gastric walls. Developing lymphatic networks represented fewer blind ends and branches than mature networks in whole-mount preparations. Many circular lymphatic-like structures exhibited VEGFR-3 expression and weak 5'-Nase activity in the early embryonic stage, showing visible morphological properties in the lymphatic endothelium. These newly formed lymphatics showed an obvious accumulation in the submucosa and serosa and a variation in the intensity of VEGFR-3 binding to endothelial cells among samples. A reaction product for anti-VEGFR-3 was found on the luminal surface of endothelial cells and on the membrane of some organelles and intraluminal lymphocytes. These findings indicate that an active proliferating feature of the clustered developing lymphatics may create a favorable environment for their sprouting and growth, which may serve as a functional requirement for lymph drainage in the region. PMID- 12588962 TI - Expression of the giant protein AHNAK (desmoyokin) in muscle and lining epithelial cells. AB - Here we report a detailed analysis of the expression and localization of the giant protein AHNAK in adult mouse tissues. We show that AHNAK is widely expressed in muscle cells, including cardiomyocytes, smooth muscle cells, skeletal muscle, myoepithelium, and myofibroblasts. AHNAK is also specifically expressed in epithelial cells of most lining epithelium, but is absent in epithelium with more specialized secretory or absorptive functions. In all adult tissues, the main localization of AHNAK is at the plasma membrane. A role for AHNAK in the specific organization and the structural support of the plasma membrane common to muscle and lining epithelium is discussed. PMID- 12588963 TI - Induction of a storage phenotype and abnormal intracellular localization of apical glycoproteins are two independent responses to GalNAcalpha-O-bn. AB - Our previous studies on an inhibitor of O-glycosylation of glycoproteins, GalNAcalpha-O-bn, in the model of enterocytic HT-29 cells, have shown at the cellular level an alteration of the normal localization of apical glycoproteins, and at the biochemical level an in situ synthesis and storage of sialylated GalNAcalpha-O-bn oligosaccharides. The purpose of this study was to examine if a relation existed between these two events, using different cell lines. Intracellular storage of GalNAcalpha-O-bn metabolites occurred in HT-29 and CAPAN 1 cells but not in Caco-2 cells. On the other hand, an accumulation of endosomal/lysosomal compartments was observed in HT-29 and CAPAN-1 cells but not in Caco-2 cells. These data focused on a GalNAcalpha-O-bn-derived storage phenotype in HT-29 and CAPAN-1 cells. The apical membrane glycoproteins MUC1 and CEA showed an abnormal localization inside intracytoplasmic vesicles in HT-29 cells, whereas they kept their normal localization in Caco-2 and CAPAN-1 cells. Studies on the glycosylation of these apical glycoproteins showed that GalNAcalpha-O-bn inhibited the glycosylation in a cell-specific manner. The alteration in the apical targeting of glycoproteins, and the appearance of a GalNAcalpha-O-bn-derived storage phenotype are two independent and cell type specific events. The former depends on the inhibition pattern of the glycosylation of endogenous glycoproteins, whereas the latter is connected to the intracellular accumulation of GalNAcalpha-O-bn metabolites. PMID- 12588964 TI - Expression of the cystathionine beta synthase (CBS) gene during mouse development and immunolocalization in adult brain. AB - Hyperhomocysteinemia, caused by a lack of cystathionine beta synthase (CBS), leads to elevated plasma concentrations of homocysteine. This is a common risk factor for atherosclerosis, stroke, and possibly neurodegenerative diseases. However, the mechanisms that link hyperhomocysteinemia due to CBS deficiency to these diseases are still unknown. Early biochemical studies describe developmental and adult patterns of transsulfuration and CBS expression in a variety of species. However, there is incomplete knowledge about the regional and cellular expression pattern of CBS, notably in the brain. To complete the previous data, we used in situ hybridization and Northern blotting to characterize the spatial and temporal patterns of Cbs gene expression during mouse development. In the early stages of development, the Cbs gene was expressed only in the liver and in the skeletal, cardiac, and nervous systems. The expression declined in the nervous system in the late embryonic stages, whereas it increased in the brain after birth, peaking during cerebellar development. In the adult brain, expression was strongest in the Purkinje cell layer and in the hippocampus. Immunohistochemical analyses showed that the CBS protein was localized in most areas of the brain but predominantly in the cell bodies and neuronal processes of Purkinje cells and Ammon's horn neurons. PMID- 12588965 TI - Temporal analysis of rat growth plates: cessation of growth with age despite presence of a physis. AB - Despite the continued presence of growth plates in aged rats, longitudinal growth no longer occurs. The aims of this study were to understand the reasons for the cessation of growth. We studied the growth plates of femurs and tibiae in Wistar rats aged 62-80 weeks and compared these with the corresponding growth plates from rats aged 2-16 weeks. During skeletal growth, the heights of the plates, especially that of the hypertrophic zone, reflected the rate of bone growth. During the period of decelerating growth, it was the loss of large hydrated chondrocytes that contributed most to the overall decrease in the heights of the growth plates. In the old rats we identified four categories of growth plate morphology that were not present in the growth plates of younger rats: (a). formation of a bone band parallel to the metaphyseal edge of the growth plate, which effectively sealed that edge; (b). extensive areas of acellularity, which were resistant to resorption and/or remodeling; (c). extensive remodeling and bone formation within cellular regions of the growth plate; and (d). direct bone formation by former growth plate chondrocytes. These processes, together with a loss of synchrony across the plate, would prevent further longitudinal expansion of the growth plate despite continued sporadic proliferation of chondrocytes. PMID- 12588966 TI - Homeobox protein, Hmx3, in postnatally developing rat submandibular glands. AB - Homeobox-containing (Hox) genes play important roles in development, particularly in the development of neurons and sensory organs, and in specification of body plan. The Hmx gene family is a new class of homeobox-containing genes defined by a conserved homeobox region and a characteristic pattern of expression in the central nervous system that is more rostral than that of the Hox genes. To date, three closely related members of the Hmx family, Hmx1, Hmx2, and Hmx3, have been described. All three Hmx genes are expressed in the craniofacial region of developing embryos. Here we show, for the first time, the expression of the transcription factor Hmx3 in postnatally developing salivary glands. Hmx3 protein is expressed in a cell type-specific manner in rat salivary glands. Hmx3 is present in both the nuclei and cytoplasm of specific groups of duct cells of the submandibular, parotid, and sublingual glands. Hmx3 expression increases during postnatal development of the submandibular gland. The duct cells show increasing concentrations of Hmx3 protein with progressive development of the submandibular gland. In contrast, the acinar cells of the three salivary glands do not exhibit detectable levels of Hmx3 protein. PMID- 12588967 TI - Unspecific labeling of pancreatic islets by antisera against fibroblast growth factors and their receptors. AB - Six distinct fibroblast growth factors (FGFs) have been detected in pancreatic islets by immunohistochemistry (IHC) using commercially available antisera. We show here that these antisera are useful for Western blotting but that only two are suited for IHC. By Western blotting, these antisera detect recombinant FGFs. Detection can be eliminated by preabsorption with immunizing peptide but not with irrelevant peptide. By IHC we find specific labeling of islets with anti-FGF1 and anti-FGF2 antisera. Labeling can be abolished by preabsorption with the immunizing peptides. In contrast, prominent staining of islets by anti-FGF4, FGF5, -FGF7, and -FGF10 antisera is unspecific because the staining cannot be competed by preabsorption with the immunizing peptides. PMID- 12588968 TI - Localization of GFP in frozen sections from unfixed mouse tissues: immobilization of a highly soluble marker protein by formaldehyde vapor. AB - Green fluorescent protein (GFP) and its variants, such as enhanced GFP (EGFP), have been introduced into mammalian cells by transgenes, e.g., to distinguish donor from host cells after transplantation. Free GFP is extremely soluble and leaks out from liquid-covered cryostat sections so that fixation of whole organs before sectioning has been mandatory. This precludes the analysis of serial sections with respect to fixation-sensitive enzyme activities and antigens. We describe here a vapor fixation for sections from unfixed cryostat blocks of tissue that allows unrestricted enzyme and immunohistochemistry on adjacent sections, as demonstrated for cross-striated muscle and other tissues from EGFP transgenic "green mice" and for a transplantation experiment. PMID- 12588969 TI - Selective removal of the selenocysteine tRNA [Ser]Sec gene (Trsp) in mouse mammary epithelium. AB - Mice homozygous for an allele encoding the selenocysteine (Sec) tRNA [Ser]Sec gene (Trsp) flanked by loxP sites were generated. Cre recombinase-dependent removal of Trsp in these mice was lethal to embryos. To investigate the role of Trsp in mouse mammary epithelium, we deleted this gene by using transgenic mice carrying the Cre recombinase gene under control of the mouse mammary tumor virus (MMTV) long terminal repeat or the whey acidic protein promoter. While both promoters target Cre gene expression to mammary epithelium, MMTV-Cre is also expressed in spleen and skin. Sec tRNA [Ser]Sec amounts were reduced by more than 70% in mammary tissue with either transgene, while in skin and spleen, levels were reduced only with MMTV-Cre. The selenoprotein population was selectively affected with MMTV-Cre in breast and skin but not in the control tissue, kidney. Moreover, within affected tissues, expression of specific selenoproteins was regulated differently and often in a contrasting manner, with levels of Sep15 and the glutathione peroxidases GPx1 and GPx4 being substantially reduced. Expression of the tumor suppressor genes BRCA1 and p53 was also altered in a contrasting manner in MMTV-Cre mice, suggesting greater susceptibility to cancer and/or increased cell apoptosis. Thus, the conditional Trsp knockout mouse allows tissue specific manipulation of Sec tRNA and selenoprotein expression, suggesting that this approach will provide a useful tool for studying the role of selenoproteins in health. PMID- 12588970 TI - Triple decoding of hepatitis C virus RNA by programmed translational frameshifting. AB - Ribosomes can be programmed to shift from one reading frame to another during translation. Hepatitis C virus (HCV) uses such a mechanism to produce F protein from the -2/+1 reading frame. We now report that the HCV frameshift signal can mediate the synthesis of the core protein of the zero frame, the F protein of the -2/+1 frame, and a 1.5-kDa protein of the -1/+2 frame. This triple decoding function does not require sequences flanking the frameshift signal and is apparently independent of membranes and the synthesis of the HCV polyprotein. Two consensus -1 frameshift sequences in the HCV type 1 frameshift signal facilitate ribosomal frameshifts into both overlapping reading frames. A sequence which is located immediately downstream of the frameshift signal and has the potential to form a double stem-loop structure can significantly enhance translational frameshifting in the presence of the peptidyl-transferase inhibitor puromycin. Based on these results, a model is proposed to explain the triple decoding activities of the HCV ribosomal frameshift signal. PMID- 12588971 TI - A methyltransferase targeting assay reveals silencer-telomere interactions in budding yeast. AB - We have designed a modified version of the Dam identification technique and used it to probe higher-order chromatin structure in Saccharomyces cerevisiae. We fused the bacterial DNA methyltransferase Dam to the DNA-binding domain of TetR and targeted the resulting chimera to Tet operators inserted in the yeast genome at the repressed locus HML. We then monitored the methylation status of HML and other sequences by a quantitative technique combining methylation-sensitive restriction and real-time PCR. As expected, we found that TetR-Dam efficiently methylated HML in cis. More strikingly, when TetR-Dam was present at HML, we observed increased methylation in the III-L subtelomeric region but not in intervening sequences. This effect was lost when the HML silencers were inactivated by mutations. When the HM silencers and the Tet operators were transferred to a plasmid, strong methylation was clearly observed not only in the III-L subtelomeric region but also at other telomeres. These data indicate that HM silencers can specifically associate with telomeres, even those located on different chromosomes. PMID- 12588972 TI - Transcript-selective translational silencing by gamma interferon is directed by a novel structural element in the ceruloplasmin mRNA 3' untranslated region. AB - Transcript-selective translational control of eukaryotic gene expression is often directed by a structural element in the 3' untranslated region (3'-UTR) of the mRNA. In the case of ceruloplasmin (Cp), induced synthesis of the protein by gamma interferon (IFN-gamma) in U937 monocytic cells is halted by a delayed translational silencing mechanism requiring the binding of a cytosolic inhibitor to the Cp 3'-UTR. Silencing requires the essential elements of mRNA circularization, i.e., eukaryotic initiation factor 4G, poly(A)-binding protein, and poly(A) tail. We here determined the minimal silencing element in the Cp 3' UTR by progressive deletions from both termini. A minimal, 29-nucleotide (nt) element was determined by gel shift assay to be sufficient for maximal binding of the IFN-gamma-activated inhibitor of translation (GAIT), an as-yet-unidentified protein or complex. The interaction was shown to be functional by an in vitro translation assay in which the GAIT element was used as a decoy to overcome translational silencing. Mutation analysis showed that the GAIT element contained a 5-nt terminal loop, a weak 3-bp helix, an asymmetric internal bulge, and a proximal 6-bp helical stem. Two invariant loop residues essential for binding activity were identified. Ligation of the GAIT element immediately downstream of a luciferase reporter conferred the translational silencing response to the heterologous transcript in vitro and in vivo; a construct containing a nonbinding, mutated GAIT element was ineffective. Translational silencing of Cp, and possibly other transcripts, mediated by the GAIT element may contribute to the resolution of the local inflammatory response following cytokine activation of macrophages. PMID- 12588974 TI - Inhibitory and stimulatory regulation of Rac and cell motility by the G12/13-Rho and Gi pathways integrated downstream of a single G protein-coupled sphingosine-1 phosphate receptor isoform. AB - The G protein-coupled receptors S1P2/Edg5 and S1P3/Edg3 both mediate sphingosine 1-phosphate (S1P) stimulation of Rho, yet S1P2 but not S1P3 mediates downregulation of Rac activation, membrane ruffling, and cell migration in response to chemoattractants. Specific inhibition of endogenous Galpha12 and Galpha13, but not of Galphaq, by expression of respective C-terminal peptides abolished S1P2-mediated inhibition of Rac, membrane ruffling, and migration, as well as stimulation of Rho and stress fiber formation. Fusion receptors comprising S1P2 and either Galpha12 or Galpha13, but not Galphaq, mediated S1P stimulation of Rho and also inhibition of Rac and migration. Overexpression of Galphai, by contrast, specifically antagonized S1P2-mediated inhibition of Rac and migration. The S1P2 actions were mimicked by expression of V14Rho and were abolished by C3 toxin and N19Rho, but not Rho kinase inhibitors. In contrast to S1P2, S1P3 mediated S1P-directed, pertussis toxin-sensitive chemotaxis and Rac activation despite concurrent stimulation of Rho via G12/13. Upon inactivation of Gi by pertussis toxin, S1P3 mediated inhibition of Rac and migration just like S1P2. These results indicate that integration of counteracting signals from the Gi- and the G12/13-Rho pathways directs either positive or negative regulation of Rac, and thus cell migration, upon activation of a single S1P receptor isoform. PMID- 12588973 TI - Mutations in the v-Rel transactivation domain indicate altered phosphorylation and identify a subset of NF-kappaB-regulated cell death inhibitors important for v-Rel transforming activity. AB - Consistent with the constitutive activation of Rel/NF-kappaB in human hematopoietic tumors, the v-Rel oncoprotein induces aggressive leukemia/lymphomas in animal models. v-Rel is thus a valuable tool to characterize the role of Rel/NF-kappaB in cancer and the mechanisms involved. Prior studies by our group identified a serine-rich domain in v-Rel that was required for biological activity. Here, we investigated the molecular basis for the transformation defect of specific serine mutants. We show that the transforming efficiency of these mutants in primary lymphoid cells is correlated with their ability to mediate kappaB site-dependent transactivation and with specific changes in phosphorylation profiles. Interestingly, coexpression of the death antagonists Bcl-xL and Bcl-2 significantly increased their oncogenicity, whereas other NF kappaB-regulated death inhibitors showed little or no effect. The fact that a subset of apoptosis inhibitors could rescue v-Rel transactivation mutants suggests that their reduced transcriptional activity may critically affect expression of defined death antagonists essential for oncogenesis. Consistent with this hypothesis, we observed selection for high endogenous expression of Bcl 2-related death antagonists in cells transformed by weakly transforming v-Rel mutants. These results emphasize the need for Rel/NF-kappaB to efficiently activate expression of a subset of antiapoptotic genes from the Bcl-2 family to manifest its oncogenic phenotype. PMID- 12588975 TI - The C terminus of initiation factor 4E-binding protein 1 contains multiple regulatory features that influence its function and phosphorylation. AB - Eukaryotic initiation factor 4E (eIF4E) binds the mRNA cap structure and forms eIF4F complexes that recruit 40S subunits to the mRNA. Formation of eIF4F is blocked by eIF4E-binding proteins such as 4E-BP1, which interacts with eIF4E via a motif in the center of its 118-residue sequence. 4E-BP1 plays key roles in cell proliferation, growth, and survival. Binding of 4E-BP1 to eIF4E is regulated by hierarchical multisite phosphorylation. Here we demonstrate that three different features in the C terminus of 4E-BP1 play distinct roles in regulating its phosphorylation and function. Firstly, we identify a new phosphorylation site in its C terminus (S101). A serine or glutamate at this position is required for efficient phosphorylation at Ser65. A second C-terminal site, S112, directly affects binding of 4E-BP1 to eIF4E without influencing phosphorylation of other sites. Thirdly, a conserved C-terminal motif influences phosphorylation of multiple residues, including rapamycin-insensitive sites. These relatively long range effects are surprising given the reportedly unstructured nature of 4E-BP1 and may imply that phosphorylation of 4E-BP1 and/or binding to eIF4E induces a more-ordered structure. 4E-BP2 and -3 lack phosphorylatable residues corresponding to both S101 and S112. However, in 4E-BP3, replacement of the alanine at the position corresponding to S112 by serine or glutamate did not confer the ability to be released from eIF4E in response to insulin. PMID- 12588976 TI - In exponentially growing Saccharomyces cerevisiae cells, rRNA synthesis is determined by the summed RNA polymerase I loading rate rather than by the number of active genes. AB - Genes encoding rRNA are multicopy and thus could be regulated by changing the number of active genes or by changing the transcription rate per gene. We tested the hypothesis that the number of open genes is limiting rRNA synthesis by using an electron microscopy method that allows direct counting of the number of active genes per nucleolus and the number of polymerases per active gene. Two strains of Saccharomyces cerevisiae were analyzed during exponential growth: a control strain with a typical number of rRNA genes ( approximately 143 in this case) and a strain in which the rRNA gene number was reduced to approximately 42 but which grows as well as controls. In control strains, somewhat more than half of the genes were active and the mean number of polymerases/gene was approximately 50 +/ 20. In the 42-copy strain, all rRNA genes were active with a mean number of 100 +/- 29 polymerases/gene. Thus, an equivalent number of polymerases was active per nucleolus in the two strains, though the number of active genes varied by twofold, showing that overall initiation rate, and not the number of active genes, determines rRNA transcription rate during exponential growth in yeast. Results also allow an estimate of elongation rate of approximately 60 nucleotides/s for yeast Pol I and a reinitiation rate of less than 1 s on the most heavily transcribed genes. PMID- 12588977 TI - Identification of p21-activated kinase specificity determinants in budding yeast: a single amino acid substitution imparts Ste20 specificity to Cla4. AB - Two closely related p21-activated kinases from Saccharomyces cerevisiae, Ste20 and Cla4, interact with and are regulated by Cdc42, a small Rho-like GTPase. These kinases are argued to perform a common essential function, based on the observation that the single mutants are viable whereas the double mutant is inviable. Despite having a common upstream regulator and at least one common function, these molecules also have many distinct cellular signaling roles. Ste20 signals upstream of several mitogen-activated protein kinase cascades (e.g., pheromone response, filamentous growth, and high osmolarity), and Cla4 signals during budding and cytokinesis. In order to investigate how these kinases are directed to distinct functions, we sought to identify specificity determinants within Ste20 and Cla4. To this end, we constructed both chimeric fusions and point mutants and tested their ability to perform unique and shared cellular roles. Specificity determinants for both kinases were mapped to the C-terminal kinase domains. Remarkably, the substitution of a single amino acid, threonine 818, from Ste20 into an otherwise wild-type Cla4, Cla4D772T, conferred the ability to perform many Ste20-specific functions. PMID- 12588978 TI - Distinct mechanisms of receptor and nonreceptor tyrosine kinase activation by reactive oxygen species in vascular smooth muscle cells: role of metalloprotease and protein kinase C-delta. AB - Reactive oxygen species (ROS) are implicated in cardiovascular diseases. ROS, such as H2O2, act as second messengers to activate diverse signaling pathways. Although H2O2 activates several tyrosine kinases, including the epidermal growth factor (EGF) receptor, JAK2, and PYK2, in vascular smooth muscle cells (VSMCs), the intracellular mechanism by which ROS activate these tyrosine kinases remains unclear. Here, we identified two distinct signaling pathways required for receptor and nonreceptor tyrosine kinase activation by H2O2 involving a metalloprotease-dependent generation of heparin-binding EGF-like growth factor (HB-EGF) and protein kinase C (PKC)-delta activation, respectively. H2O2-induced EGF receptor tyrosine phosphorylation was inhibited by a metalloprotease inhibitor, whereas the inhibitor had no effect on H2O2-induced JAK2 tyrosine phosphorylation. HB-EGF neutralizing antibody inhibited H2O2-induced EGF receptor phosphorylation. In COS-7 cells expressing an HB-EGF construct tagged with alkaline phosphatase, H2O2 stimulates HB-EGF production through metalloprotease activation. By contrast, dominant negative PKC-delta transfection inhibited H2O2 induced JAK2 phosphorylation but not EGF receptor phosphorylation. Dominant negative PYK2 inhibited H2O2-induced JAK2 activation but not EGF receptor activation, whereas dominant negative PKC-delta inhibited PYK2 activation by H2O2. These data demonstrate the presence of distinct tyrosine kinase activation pathways (PKC-delta/PYK2/JAK2 and metalloprotease/HB-EGF/EGF receptor) utilized by H2O2 in VSMCs, thus providing unique therapeutic targets for cardiovascular diseases. PMID- 12588979 TI - Phosphorylation of stem-loop binding protein (SLBP) on two threonines triggers degradation of SLBP, the sole cell cycle-regulated factor required for regulation of histone mRNA processing, at the end of S phase. AB - The replication-dependent histone mRNAs, the only eukaryotic mRNAs that do not have poly(A) tails, are present only in S-phase cells. Coordinate posttranscriptional regulation of histone mRNAs is mediated by the stem-loop at the 3' end of histone mRNAs. The protein that binds the 3' end of histone mRNA, stem-loop binding protein (SLBP), is required for histone pre-mRNA processing and is involved in multiple aspects of histone mRNA metabolism. SLBP is also regulated during the cell cycle, accumulating as cells enter S phase and being rapidly degraded as cells exit S phase. Mutation of any residues in a TTP sequence (amino acids 60 to 62) or mutation of a consensus cyclin binding site (amino acids 99 to 104) stabilizes SLBP in G2 and mitosis. These two threonines are phosphorylated in late S phase, as determined by mass spectrometry (MS) of purified SLBP from late S-phase cells, triggering SLBP degradation. Cells that express a stable SLBP still degrade histone mRNA at the end of S phase, demonstrating that degradation of SLBP is not required for histone mRNA degradation. Nuclear extracts from G1 and G2 cells are deficient in histone pre mRNA processing, which is restored by addition of recombinant SLBP, indicating that SLBP is the only cell cycle-regulated factor required for histone pre-mRNA processing. PMID- 12588981 TI - Balance between acetylation and methylation of histone H3 lysine 9 on the E2F responsive dihydrofolate reductase promoter. AB - Epigenetic marks that specify silent heterochromatic domains in eucaryotic genomes include methylation of histone H3 lysine 9. Strikingly, active loci in the vicinity of silent domains are sometimes characterized by acetylation of histone H3 lysine 9, suggesting that the balance between these two competitive modifications is important for the establishment of specific chromatin structures. Some euchromatic genes, targeted by the retinoblastoma protein Rb, are also believed to be regulated by histone H3 lysine 9 methylation. Here, we study the dihydrofolate reductase promoter, which is repressed in G0 and at the beginning of G1 by p107 or p130, two Rb-related proteins. We found that these two pocket proteins share with Rb the ability to associate with the histone methyl transferase SUV39H1. SUV39H1 can be recruited to the E2F transcription factor and functions as a transcriptional corepressor. With ChIP assays followed by real time PCR, we showed that K9 of histone H3 evolves from a hypermethylated state in G0 to a hyperacetylated state at the G1/S transition. Taken together, these results indicate that the temporal regulation of euchromatic promoters may involve controlling the balance between methylation and acetylation of histone H3 lysine 9, a feature previously described for the spatial regulation of chromatin function. PMID- 12588980 TI - Delayed rRNA processing results in significant ribosome biogenesis and functional defects. AB - mof6-1 was originally isolated as a recessive mutation in Saccharomyces cerevisiae which promoted increased efficiencies of programmed -1 ribosomal frameshifting and rendered cells unable to maintain the killer virus. Here, we demonstrate that mof6-1 is a unique allele of the histone deacetylase RPD3, that the deacetylase function of Rpd3p is required for controlling wild-type levels of frameshifting and virus maintenance, and that the closest human homolog can fully complement these defects. Loss of the Rpd3p-associated histone deacetylase function, either by mutants of rpd3 or loss of the associated gene product Sin3p or Sap30p, results in a delay in rRNA processing rather than in an rRNA transcriptional defect. This results in production of ribosomes having lower affinities for aminoacyl-tRNA and diminished peptidyltransferase activities. We hypothesize that decreased rates of peptidyl transfer allow ribosomes with both A and P sites occupied by tRNAs to pause for longer periods of time at -1 frameshift signals, promoting increased programmed -1 ribosomal frameshifting efficiencies and subsequent loss of the killer virus. The frameshifting defect is accentuated when the demand for ribosomes is highest, suggesting that rRNA posttranscriptional modification is the bottleneck in ribosome biogenesis. PMID- 12588982 TI - Collaborative competition mechanism for gene activation in vivo. AB - The mechanism by which gene regulatory proteins gain access to their DNA target sites is not known. In vitro, binding is inherently cooperative between arbitrary DNA binding proteins whose target sites are located within the same nucleosome. We refer to such competition-based cooperativity as collaborative competition. Here we show that arbitrarily chosen foreign DNA binding proteins, LexA and Tet repressor, cooperate with an adjacently binding endogenous activator protein, Gcn4, to coactivate expression of chromosomal reporter genes in Saccharomyces cerevisiae. Coactivation requires that the cooperating target sites be within a nucleosome-length distance; it leads to increased occupancy by Gcn4 at its binding site; and it requires both Gcn5 and Swi/Snf which, at an endogenous Gcn4 dependent promoter, act subsequent to Gcn4 binding. These results imply that collaborative competition contributes to gene regulation in vivo. They further imply that, even in the presence of the cell's full wild-type complement of chromatin remodeling factors, competition of regulatory proteins with histone octamer for access to regulatory target sites remains a quantitative determinant of gene expression levels. We speculate that initial target site recognition and binding may occur via spontaneous nucleosomal site exposure, with remodeling factor action required downstream to lock in higher levels of regulatory protein occupancy. PMID- 12588983 TI - Identification of a structural determinant necessary for the localization and function of estrogen receptor alpha at the plasma membrane. AB - Estrogen receptors (ER) have been localized to the cell plasma membrane (PM), where signal transduction mediates some estradiol (E2) actions. However, the precise structural features of ER that result in membrane localization have not been determined. We obtained a partial tryptic peptide/mass spectrometry analysis of membrane mouse ERalpha protein. Based on this, we substituted alanine for the determined serine at amino acid 522 within the E domain of wild-type (wt) ERalpha. Upon transfection in CHO cells, the S522A mutant ERalpha resulted in a 62% decrease in membrane receptor number and reduced colocalization with caveolin 1 relative to those with expression of wt ERalpha. E2 was significantly less effective in stimulating multiple rapid signals from the membranes of CHO cells expressing ERalpha S522A than from those of CHO cells expressing wt ERalpha. In contrast, nuclear receptor expression and transcriptional function were very similar. The S522A mutant was also 60% less effective than wt ERalpha in binding caveolin 1, which facilitates ER transport to the PM. All functions of ERalpha mutants with other S-to-A substitutions were comparable to those of wt ER, and deletion of the A/B or C domain had little consequence for membrane localization or function. Transfection of ERalpha S522A into breast cancer cells that express native ER downregulated E2 binding at the membrane, signaling to ERK, and G1/S cell cycle events and progression. However, there was no effect on the E2 transactivation of an ERE-luciferase reporter. In summary, serine 522 is necessary for the efficient translocation and function of ERalpha at the PM. The S522A mutant also serves as a dominant-negative construct, identifying important functions of E2 that originate from activating PM ER. PMID- 12588984 TI - YPXL/I is a protein interaction motif recognized by aspergillus PalA and its human homologue, AIP1/Alix. AB - The zinc finger transcription factor PacC undergoes two-step proteolytic activation in response to alkaline ambient pH. PalA is a component of the fungal ambient pH signal transduction pathway. Its mammalian homologue AIP1/Alix interacts with the apoptosis-linked protein ALG-2. We show that both PalA and AIP1/Alix recognize a protein-protein binding motif that we denote YPXL/I, where Tyr, Pro, and Leu/Ile are crucial for its interactive properties. Two such motifs flanking the signaling protease cleavage site mediate direct binding of PalA to PacC, required for the first and only pH-regulated cleavage of this transcription factor. PalA can bind the "closed" (i.e., wild-type full-length) conformer of PacC, suggesting that PalA binding constitutes the first stage in the two-step proteolytic cascade, recruiting or facilitating access of the signaling protease, presumably PalB. In addition to recognizing YPXL/I motifs, both PalA and AIP1/Alix interact with the Aspergillus class E Vps protein Vps32 homologue, a member of a protein complex involved in the early steps of the multivesicular body pathway, suggesting that this interaction is an additional feature of proteins of the PalA/AIP1/Alix family. PMID- 12588985 TI - MBDin, a novel MBD2-interacting protein, relieves MBD2 repression potential and reactivates transcription from methylated promoters. AB - We have identified a human gene encoding a novel MBD2-interacting protein (MBDin) that contains an N-terminal GTP-binding site, a putative nuclear export signal (NES), and a C-terminal acidic region. MBDin cDNA was isolated through a two hybrid interaction screening using the methyl-CpG-binding protein MBD2 as bait. The presence of the C-terminal 46-amino-acid region of MBD2 and both the presence of the acidic C-terminal 128-amino-acid region and the integrity of the GTP binding site of MBDin were required for the interaction. Interaction between MBD2 and MBDin in mammalian cells was confirmed by immunoprecipitation experiments. Fluorescence imaging experiments demonstrated that MBDin mainly localizes in the cytoplasm but accumulates in the nucleus upon disruption of the NES or treatment with leptomycin B, an inhibitor of NES-mediated transport. We also found that MBDin partially colocalizes with MBD2 at foci of heavily methylated satellite DNA. An MBD2 deletion mutant lacking the C-terminal region maintained its subnuclear localization but failed to recruit MBDin at hypermethylated foci. Functional analyses demonstrated that MBDin relieves MBD2-mediated transcriptional repression both when Gal4 chimeric constructs and when in vitro methylated promoter-reporter plasmids were used in transcriptional assays. Southern blotting and bisulfite analysis showed that transcriptional reactivation occurred without changes of the promoter methylation pattern. Our findings suggest the existence of factors that could be targeted on methylated DNA by methyl-CpG-binding proteins reactivating transcription even prior to demethylation. PMID- 12588986 TI - BRCT domain-containing protein PTIP is essential for progression through mitosis. AB - The Pax transactivation domain-interacting protein (PTIP) is a large nuclear protein with multiple BRCT domains that was identified on the basis of its interaction with transcription factors of the Pax and Smad families. To address the function of PTIP during mouse development, we generated a constitutive null allele. Homozygous PTIP mutants are developmentally retarded, disorganized, and embryonic lethal by day 9.5 of embryonic development (E9.5). PTIP mutant cells appear to replicate DNA but show reduced levels of mitosis and widespread cell death by E8.5. DNA damage appears to precede nuclear condensation at E7.5, suggesting a defect in DNA repair. Neither embryonic fibroblast nor embryonic stem cells from PTIP mutants proliferate in culture, suggesting a fundamental defect in cell proliferation. Trophoblast cells from PTIP mutants are more sensitive to DNA-damaging agents. Condensation of chromatin and expression of phospho-histone H3 are also affected in PTIP mutants, and this may underlie the inability of PTIP mutants to progress through mitosis. Given the role of BRCT domain proteins in DNA repair and cell cycle control, we propose that PTIP is an essential element of the cell proliferation machinery, perhaps by functioning in the DNA repair pathways. PMID- 12588987 TI - Beta-catenin binds to the activation function 2 region of the androgen receptor and modulates the effects of the N-terminal domain and TIF2 on ligand-dependent transcription. AB - Beta-catenin is a multifunctional molecule that is activated by signaling through WNT receptors. beta-Catenin can also enhance the transcriptional activity of some steroid hormone receptors such as the androgen receptor and retinoic acid receptor alpha. Androgens can affect nuclear translocation of beta-catenin and influence its subcellular distribution. Using mammalian two-hybrid binding assays, analysis of reporter gene transcription, and coimmunoprecipitation, we now show that beta-catenin binds to the androgen receptor ligand-binding domain (LBD) and modulates the transcriptional effects of TIF2 and the androgen receptor N-terminal domain (NTD). In functional assays, beta-catenin bound to androgen receptor only in the presence of ligand agonists, not antagonists. Beta-catenin binding to the androgen receptor LBD was independent of and cooperative with the androgen receptor NTD and the p160 coactivator TIF2, both of which bind to the activation function 2 (AF-2) region of the androgen receptor. Different mutations of androgen receptor helix 3 amino acids disrupted binding of androgen receptor NTD and beta-catenin. beta-Catenin, androgen receptor NTD, and TIF2 binding to the androgen receptor LBD were affected similarly by a subset of helix 12 mutations, but disruption of two sites on helix 12 affected only binding of beta catenin and not of TIF2 or the androgen receptor NTD. Mutational disruption of each of five LXXLL peptide motifs in the beta-catenin armadillo repeats did not disrupt either binding to androgen receptor or transcriptional coactivation. ICAT, an inhibitor of T-cell factor 4 (TCF-4), and E-cadherin binding to beta catenin also blocked binding of the androgen receptor LBD. We also demonstrated cross talk between the WNT and androgen receptor signaling pathways because excess androgen receptor could interfere with WNT signaling and excess TCF-4 inhibited the interaction of beta-catenin and androgen receptor. Taken together, the data show that beta-catenin can bind to the androgen receptor LBD and modulate the effects of the androgen receptor NTD and TIF2 on transcription. PMID- 12588989 TI - Biphasic activation of Aurora-A kinase during the meiosis I- meiosis II transition in Xenopus oocytes. AB - Xenopus Aurora-A (also known as Eg2) is a member of the Aurora family of mitotic serine/threonine kinases. In Xenopus oocytes, Aurora-A phosphorylates and activates a cytoplasmic mRNA polyadenylation factor (CPEB) and therefore plays a pivotal role in MOS translation. However, hyperphosphorylation and activation of Aurora-A appear to be dependent on maturation-promoting factor (MPF) activation. To resolve this apparent paradox, we generated a constitutively activated Aurora A by engineering a myristylation signal at its N terminus. Injection of Myr Aurora-A mRNA induced germinal vesicle breakdown (GVBD) with the concomitant activation of MOS, mitogen-activated protein kinase, and MPF. Myr-Aurora-A injected oocytes, however, appeared to arrest in meiosis I with high MPF activity and highly condensed, metaphase-like chromosomes but no organized microtubule spindles. No degradation of CPEB or cyclin B2 was observed following GVBD in Myr Aurora-A-injected oocytes. In the presence of progesterone, the endogenous Aurora A became hyperphosphorylated and activated at the time of MPF activation. Following GVBD, Aurora-A was gradually dephosphorylated and inactivated before it was hyperphosphorylated and activated again. This biphasic pattern of Aurora-A activation mirrored that of MPF activation and hence may explain meiosis I arrest by the constitutively activated Myr-Aurora-A. PMID- 12588988 TI - The growth factor granulin interacts with cyclin T1 and modulates P-TEFb dependent transcription. AB - Cyclin T1, together with the kinase CDK9, is a component of the transcription elongation factor P-TEFb which binds the human immunodeficiency virus type 1 (HIV 1) transactivator Tat. P-TEFb facilitates transcription by phosphorylating the carboxy-terminal domain (CTD) of RNA polymerase II. Cyclin T1 is an exceptionally large cyclin and is therefore a candidate for interactions with regulatory proteins. We identified granulin as a cyclin T1-interacting protein that represses expression from the HIV-1 promoter in transfected cells. The granulins, mitogenic growth factors containing repeats of a cysteine-rich motif, were reported previously to interact with Tat. We show that granulin formed stable complexes in vivo and in vitro with cyclin T1 and Tat. Granulin bound to the histidine-rich domain of cyclin T1, which was recently found to bind to the CTD, but not to cyclin T2. Binding of granulin to P-TEFb inhibited the phosphorylation of a CTD peptide. Granulin expression inhibited Tat transactivation, and tethering experiments showed that this effect was due, at least in part, to a direct action on cyclin T1 in the absence of Tat. In addition, granulin was a substrate for CDK9 but not for the other transcription-related kinases CDK7 and CDK8. Thus, granulin is a cellular protein that interacts with cyclin T1 to inhibit transcription. PMID- 12588990 TI - The N-terminal 24 amino acids of the p55 gamma regulatory subunit of phosphoinositide 3-kinase binds Rb and induces cell cycle arrest. AB - Although phosphoinositide 3-kinase (PI 3-kinase) is essential for cell cycle progression, the molecular mechanisms that regulate its diverse biological effects are poorly understood. We demonstrate here that Rb, a key regulator of cell cycle progression, associates with p55 kDa (p55alpha and p55gamma) regulatory subunits of PI 3-kinase in vivo and in vitro. Both confocal microscopy and biochemical analysis demonstrated the presence of p55gamma in the nucleus. The 24-amino-acid N-terminal end of p55gamma, which is unique among PI 3-kinase regulatory subunits, was sufficient to bind Rb. Addition of serum or growth factors to quiescent cells triggered the dissociation of Rb from p55. Ectopic expression of the 24-amino-acid N-terminal end of p55gamma inhibited cell cycle progression, as evidenced by induction of cell growth arrest at the G0/G1 phase, inhibition of DNA synthesis, inhibition of cyclin D and cyclin E promoter activity, and changes in the expression of cell cycle-related proteins. The inhibitory effects of the N-terminal end of p55gamma on cell cycle progression depended on the presence of functional Rb. These data demonstrate for the first time an association of p55gamma with Rb and show that modification of this association can lead to cell cycle arrest. PMID- 12588991 TI - Karyopherin-mediated nuclear import of the homing endonuclease VMA1-derived endonuclease is required for self-propagation of the coding region. AB - VMA1-derived endonuclease (VDE), a site-specific endonuclease in Saccharomyces cerevisiae, enters the nucleus to generate a double-strand break in the VDE negative allelic locus, mediating the self-propagating gene conversion called homing. Although VDE is excluded from the nucleus in mitotic cells, it relocalizes at premeiosis, becoming localized in both the nucleus and the cytoplasm in meiosis. The nuclear localization of VDE is induced by inactivation of TOR kinases, which constitute central regulators of cell differentiation in S. cerevisiae, and by nutrient depletion. A functional genomic approach revealed that at least two karyopherins, Srp1p and Kap142p, are required for the nuclear localization pattern. Genetic and physical interactions between Srp1p and VDE imply direct involvement of karyopherin-mediated nuclear transport in this process. Inactivation of TOR signaling or acquisition of an extra nuclear localization signal in the VDE coding region leads to artificial nuclear localization of VDE and thereby induces homing even during mitosis. These results serve as evidence that VDE utilizes the host systems of nutrient signal transduction and nucleocytoplasmic transport to ensure the propagation of its coding region. PMID- 12588992 TI - Asymmetric recognition of nonconsensus AP-1 sites by Fos-Jun and Jun-Jun influences transcriptional cooperativity with NFAT1. AB - Many regulatory elements in eukaryotic promoters do not correspond to optimal recognition sequences for the transcription factors that regulate promoter function by binding to the elements. The sequence of the binding site may influence the structural and functional properties of regulatory protein complexes. Fos-Jun heterodimers were found to bind nonconsensus AP-1 sites in a preferred orientation. Oriented Fos-Jun heterodimer binding was attributed to nonidentical recognition of the two half-sites by Fos and Jun. Jun bound preferentially to the consensus half-site, whereas Fos was able to bind nonconsensus half-sites. The orientation of heterodimer binding affected the transcriptional cooperativity of Fos-Jun-NFAT1 complexes at composite regulatory elements in mammalian cells. Jun dimerization with Fos versus ATF2 caused it to bind opposite half-sites at nonconsensus AP-1 elements. Similarly, ATF2 bound to opposite half-sites in Fos-ATF2-NFAT1 and ATF2-Jun-NFAT1 complexes. The orientations of nonconsensus AP-1 sites within composite regulatory elements affected the cooperativity of Fos-Jun as well as Jun-Jun binding with NFAT1. Since Jun homodimers cannot bind to AP-1 sites in a preferred orientation, the effects of the orientations of nonconsensus AP-1 sites on the stabilities of Jun Jun-NFAT1 complexes are likely to be due to asymmetric conformational changes in the two subunits of the homodimer. Nonconsensus AP-1 site orientation also affected the synergy of transcription activation between Jun homodimers and NFAT1 at composite regulatory elements. The asymmetric recognition of nonconsensus AP-1 sites can therefore influence the transcriptional activities of Fos and Jun both through effects on the orientation of heterodimer binding and through differential conformational changes in the two subunits of the dimer. PMID- 12588993 TI - Far3 and five interacting proteins prevent premature recovery from pheromone arrest in the budding yeast Saccharomyces cerevisiae. AB - In budding yeast, diffusible mating pheromones initiate a signaling pathway that culminates in several responses, including cell cycle arrest. Only a handful of genes required for the interface between pheromone response and the cell cycle have been identified, among them FAR1 and FAR3; of these, only FAR1 has been extensively characterized. In an effort to learn about the mechanism by which Far3 acts, we used the two-hybrid method to identify interacting proteins. We identified five previously uncharacterized open reading frames, dubbed FAR7, FAR8, FAR9, FAR10, and FAR11, that cause a far3-like pheromone arrest defect when disrupted. Using two-hybrid and coimmunoprecipitation analysis, we found that all six Far proteins interact with each other. Moreover, velocity sedimentation experiments suggest that Far3 and Far7 to Far11 form a complex. The phenotype of a sextuple far3far7-far11 mutant is no more severe than any single mutant. Thus, FAR3 and FAR7 to FAR11 all participate in the same pathway leading to G1 arrest. These mutants initially arrest in response to pheromone but resume budding after 10 h. Under these conditions, wild-type cells fail to resume budding even after several days whereas far1 mutant cells resume budding within 1 h. We conclude that the FAR3-dependent arrest pathway is functionally distinct from that which employs FAR1. PMID- 12588994 TI - Hsc70 regulates accumulation of cyclin D1 and cyclin D1-dependent protein kinase. AB - The cyclin D-dependent kinase is a critical mediator of mitogen-dependent G1 phase progression in mammalian cells. Given the high incidence of cyclin D1 overexpression in human neoplasias, the nature and complexity of cyclin D complexes in vivo have been subjects of intense interest. Besides its catalytic partner, the nature and complexity of cyclin D complexes in vivo remain ambiguous. To address this issue, we purified native cyclin D1 complexes from proliferating mouse fibroblasts by affinity chromatography and began to identify and functionally characterize the associated proteins. In this report, we describe the identification of Hsc70 and its functional importance for cyclin D1 and cyclin D1-dependent kinase maturation. We demonstrate that Hsc70 associates with newly synthesized cyclin D1 and is a component of a mature, catalytically active cyclin D1/CDK4 holoenzyme complex. Our data suggest that Hsc70 promotes stabilization of newly synthesized cyclin D1, thereby increasing its availability for assembly with CDK4. In addition, our data demonstrate that Hsc70 remains bound to cyclin D1 following its assembly with CDK4 and Cip/Kip proteins, where it ensures the formation of a catalytically active complex. PMID- 12588995 TI - War1p, a novel transcription factor controlling weak acid stress response in yeast. AB - The Saccharomyces cerevisiae ATP-binding cassette (ABC) transporter Pdr12p effluxes weak acids such as sorbate and benzoate, thus mediating stress adaptation. In this study, we identify a novel transcription factor, War1p, as the regulator of this stress adaptation through transcriptional induction of PDR12. Cells lacking War1p are weak acid hypersensitive, since they fail to induce Pdr12p. The nuclear Zn2Cys6 transcriptional regulator War1p forms homodimers and is rapidly phosphorylated upon sorbate stress. The appearance of phosphorylated War1p isoforms coincides with transcriptional activation of PDR12. Promoter deletion analysis identified a novel cis-acting weak acid response element (WARE) in the PDR12 promoter required for PDR12 induction. War1p recognizes and decorates the WARE both in vitro and in vivo, as demonstrated by band shift assays and in vivo footprinting. Importantly, War1p occupies the WARE in the presence and absence of stress, demonstrating constitutive DNA binding in vivo. Our results suggest that weak acid stress triggers phosphorylation and perhaps activation of War1p. In turn, War1p activation is necessary for the induction of PDR12 through a novel signal transduction event that elicits weak organic acid stress adaptation. PMID- 12588996 TI - Overexpression of kinase suppressor of Ras upregulates the high-molecular-weight tropomyosin isoforms in ras-transformed NIH 3T3 fibroblasts. AB - The down-regulation of the high-molecular-weight isoforms of tropomyosin (TM) is considered to be an essential event in cellular transformation. In ras transformed fibroblasts, the suppression of TM is dependent on the activity of the Raf-1 kinase; however, the requirement for other downstream effectors of Ras, such as MEK and ERK, is less clear. In this study, we have utilized the mitogen activated protein kinase scaffolding protein Kinase Suppressor of Ras (KSR) to further investigate the regulation of TM and to clarify the importance of MEK/ERK signaling in this process. Here, we report that overexpression of wild-type KSR1 in ras-transformed fibroblasts restores TM expression and induces cell flattening and stress fiber formation. Moreover, we find that the transcriptional activity of a TM-alpha promoter is decreased in ras-transformed cells and that the restoration of TM by KSR1 coincides with increased transcription from this promoter. Although ERK activity was suppressed in cells overexpressing KSR1, ERK inhibition alone was insufficient to upregulate TM expression. The KSR1-mediated effects on stress fiber formation and TM transcription required the activity of the ROCK kinase, because these effects could be suppressed by the ROCK inhibitor, Y27632. Overexpression of KSR1 did not directly regulate ROCK activity, but did permit the recoupling of ROCK to the actin polymerization machinery. Finally, all of the KSR1-induced effects were mediated by the C-terminal domain of KSR1 and were dependent on the KSR-MEK interaction. PMID- 12588997 TI - Nob1p is required for cleavage of the 3' end of 18S rRNA. AB - We report the characterization of a novel factor, Nob1p (Yor056c), which is essential for the synthesis of 40S ribosome subunits. Genetic depletion of Nob1p strongly inhibits the processing of the 20S pre-rRNA to the mature 18S rRNA, leading to the accumulation of high levels of the 20S pre-rRNA together with novel degradation intermediates. 20S processing occurs within a pre-40S particle after its export from the nucleus to the cytoplasm. Consistent with a direct role in this cleavage, Nob1p was shown to be associated with the pre-40S particle and to be present in both the nucleus and the cytoplasm. This suggests that Nob1p accompanies the pre-40S ribosomes during nuclear export. Pre-40S export is not, however, inhibited by depletion of Nob1p. PMID- 12588998 TI - Cascade of distinct histone modifications during collagenase gene activation. AB - Gene activation in eukaryotes requires chromatin remodeling, in part via histone modifications. To study the events at the promoter of a mitogen-inducible gene, we examined the induction of expression of the collagenase gene. It has been established that the collagenase gene can be activated by c-Jun and c-Fos and that the transcriptional coactivator p300 is involved in the activation. As expected, we found histone acetyltransferase activity at the collagenase promoter during activation. Interestingly, we also found histone methyltransferase and kinase activity. Strikingly, the first modification observed is methylation of histone H3 lysine 4, which correlates with the binding of the SET9 methyltransferase and the assembly of a complex consisting of c-Jun, c-Fos, TATA binding protein, and RNA polymerase II. The assembly of the preinitiation complex also shows an ordered binding of the acetyltransferase p300, the RSK2 kinase, and the SWI/SNF component Brg-1. Our results suggest that collagenase gene activation involves a dynamic recruitment of different factors and that in addition to acetylation, histone H3 lysine 4 di- and trimethylation and histone H3 serine 10 phosphorylation are important steps in the activation of this gene. PMID- 12588999 TI - A mutant receptor tyrosine phosphatase, CD148, causes defects in vascular development. AB - Vascularization defects in genetic recombinant mice have defined critical roles for a number of specific receptor tyrosine kinases. Here we evaluated whether an endothelium-expressed receptor tyrosine phosphatase, CD148 (DEP-1/PTPeta), participates in developmental vascularization. A mutant allele, CD148(DeltaCyGFP), was constructed to eliminate CD148 phosphatase activity by in frame replacement of cytoplasmic sequences with enhanced green fluorescent protein sequences. Homozygous mutant mice died at midgestation, before embryonic day 11.5 (E11.5), with vascularization failure marked by growth retardation and disorganized vascular structures. Structural abnormalities were observed as early as E8.25 in the yolk sac, prior to the appearance of intraembryonic defects. Homozygous mutant mice displayed enlarged vessels comprised of endothelial cells expressing markers of early differentiation, including VEGFR2 (Flk1), Tal1/SCL, CD31, ephrin-B2, and Tie2, with notable lack of endoglin expression. Increased endothelial cell numbers and mitotic activity indices were demonstrated. At E9.5, homozygous mutant embryos showed homogeneously enlarged primitive vessels defective in vascular remodeling and branching, with impaired pericyte investment adjacent to endothelial structures, in similarity to endoglin-deficient embryos. Developing cardiac tissues showed expanded endocardial projections accompanied by defective endocardial cushion formation. These findings implicate a member of the receptor tyrosine phosphatase family, CD148, in developmental vascular organization and provide evidence that it regulates endothelial proliferation and endothelium-pericyte interactions. PMID- 12589000 TI - Multiple functional elements comprise a Mammalian chromosomal replicator. AB - The structure of replication origins in metazoans is only nominally similar to that in model organisms, such as Saccharomyces cerevisiae. By contrast to the compact origins of budding yeast, in metazoans multiple elements act as replication start sites or control replication efficiency. We first reported that replication forks diverge from an origin 5' to the human c-myc gene and that a 2.4-kb core fragment of the origin displays autonomous replicating sequence activity in plasmids and replicator activity at an ectopic chromosomal site. Here we have used clonal HeLa cell lines containing mutated c-myc origin constructs integrated at the same chromosomal location to identify elements important for DNA replication. Replication activity was measured before or after integration of the wild-type or mutated origins using PCR-based nascent DNA abundance assays. We find that deletions of several segments of the c-myc origin, including the DNA unwinding element and transcription factor binding sites, substantially reduced replicator activity, whereas deletion of the c-myc promoter P1 had only a modest effect. Substitution mutagenesis indicated that the sequence of the DNA unwinding element, rather than the spacing of flanking sequences, is critical. These results identify multiple functional elements essential for c-myc replicator activity. PMID- 12589001 TI - IGF-I isoforms and ageing skeletal muscle: an 'unresponsive' hypertrophy agent? PMID- 12589004 TI - Coherent quantum dynamics of a superconducting flux qubit. AB - We have observed coherent time evolution between two quantum states of a superconducting flux qubit comprising three Josephson junctions in a loop. The superposition of the two states carrying opposite macroscopic persistent currents is manipulated by resonant microwave pulses. Readout by means of switching-event measurement with an attached superconducting quantum interference device revealed quantum-state oscillations with high fidelity. Under strong microwave driving, it was possible to induce hundreds of coherent oscillations. Pulsed operations on this first sample yielded a relaxation time of 900 nanoseconds and a free induction dephasing time of 20 nanoseconds. These results are promising for future solid-state quantum computing. PMID- 12589005 TI - Magmatic dike resonances inferred from very-long-period seismic signals. AB - Very-long-period (VLP) signals showing simple decaying harmonic oscillations with periods near 10 seconds and lasting for about 300 seconds were observed in association with an earthquake swarm that occurred beneath Hachijo Island, Japan. Results from the source-mechanism analysis and waveform simulation based on a fluid-filled crack model consistently point to the resonance of a dike filled with a basaltic magma as the source of the VLP signals. Thus, VLP signals can be used to probe the state of the fluid and dynamic processes within a volcanic system. PMID- 12589003 TI - Spread of HTLV-I between lymphocytes by virus-induced polarization of the cytoskeleton. AB - Cell contact is required for efficient transmission of human T cell leukemia virus- type 1 (HTLV-I) between cells and between individuals, because naturally infected lymphocytes produce virtually no cell-free infectious HTLV-I particles. However, the mechanism of cell-to-cell spread of HTLV-I is not understood. We show here that cell contact rapidly induces polarization of the cytoskeleton of the infected cell to the cell-cell junction. HTLV-I core (Gag protein) complexes and the HTLV-I genome accumulate at the cell-cell junction and are then transferred to the uninfected cell. Other lymphotropic viruses, such as HIV-1, may similarly subvert normal T cell physiology to allow efficient propagation between cells. PMID- 12589006 TI - Epilepsy: time for review. PMID- 12589007 TI - Clinical advances in therapies targeting the interleukin-2 receptor. PMID- 12589008 TI - Comparative efficacy of thrombolytics in acute myocardial infarction: a systematic review. AB - BACKGROUND: The comparative clinical effectiveness of new (reteplase, tenecteplase) vs. older (alteplase, streptokinase) thrombolytic agents in the treatment of acute myocardial infarction is uncertain. AIM: To examine 30-35 day mortality and major adverse effects of thrombolytic agents in the treatment of acute myocardial infarction. DESIGN: Systematic review of randomized controlled trials comparing the clinical efficacy of included drug regimens. METHODS: We searched MEDLINE, EMBASE, Science Citation Index/Web of Science from 1980 to December 2001, and the Cochrane Library (2001, Issue 4). Reference lists of included studies and a number of medical journals were hand searched. Randomized controlled trials that compared any two of the included drugs provided to patients in the early stages of acute myocardial infarction, were included. Outcome measures included: mortality, bleeding, stroke, reinfarction, allergy and anaphylaxis. RESULTS: We found 14 studies, total study population 142 907. For available comparisons (all alteplase vs. streptokinase, reteplase vs. streptokinase or alteplase, tenecteplase vs. alteplase), meta-analysis showed no significant differences in mortality at 30-35 days. The GUSTO-I study showed an apparent benefit of accelerated alteplase over streptokinase, but its inclusion or exclusion made little difference. Total stroke and haemorrhagic stroke rates were lower for streptokinase than for all alteplase combined (total stroke, OR 1.29, 95%CI 1.13-1.46; haemorrhagic stroke OR 1.83, 95%CI 1.14-2.93). DISCUSSION: All thrombolytic drugs appear to be of similar efficacy in reducing mortality, and the apparent benefits of accelerated alteplase in GUSTO-I are consistent with this. Whether accelerated alteplase is sufficiently different from other regimens of administering alteplase to be excluded from a meta-analysis, and whether more weight should be placed on a meta-analysis than on a single trial, are matters for debate. PMID- 12589009 TI - Characteristics of patients presenting to a cardiac clinic with palpitation. AB - BACKGROUND: Palpitation is a very common presenting symptom in primary care and in cardiac clinics, associated with marked disability. Although serious arrhythmias are uncommon causes, treatment of persistent palpitation is difficult. AIM: To describe the cardiological, behavioural and psychological characteristics of consecutive patients presenting to a cardiac clinic with the main complaint of palpitation. DESIGN: Prospective evaluation of consecutive out patients. METHODS: Participants were 184 consecutive patients with the complaint of palpitation referred to an out-patient cardiac clinic. Three assessments were used. Three to four weeks prior to clinic attendance, measures of symptoms, distress and disability were gathered, and a heart rate perception test was conducted. At the out-patient clinic, a routine clinical assessment was made. Three months later, patients received a questionnaire which included baseline measures of symptoms, distress and disability. RESULTS: Palpitation was associated with arrhythmias in 62 patients (34%), extrasystoles in 75 patients (41%) and awareness of sinus rhythm in 47 patients (26%). Distress and disability were common and persistent. There were significant differences in the characteristics of the three groups. DISCUSSION: Most patients presenting to secondary care with palpitation do not have serious underlying cardiovascular conditions. Concurrent psychological problems are common and persistent. Aetiology may be seen as an interaction of pathology, awareness of normal physiology, and psychological variables. Few patients require specialist cardiological treatment, but simple reassurance is of limited effectiveness. A stepped care approach may improve outcomes and needs rigorous evaluation. PMID- 12589010 TI - Changes in paracetamol, antidepressants and opioid poisoning in Scotland during the 1990s. AB - BACKGROUND: Overdose is one of the commonest causes of medical admissions to UK hospitals. In Scotland (pop. 5.1 million), all NHS hospital discharge data is uniquely linked to enable identification of individuals re-presenting with the same diagnosis. AIM: To examine trends in discharges for poisoning, in particular paracetamol, antidepressants and opioids from 1990-99. DESIGN: Retrospective analysis. METHODS: Discharge data from the Scottish Morbidity Record (SMR01) and mortality data from the General Register Office for Scotland (GROS) were analysed for 1990-99 by age and gender for the relevant codes. RESULTS: Overall discharge rates increased until 1997, after which they fell. This pattern was seen in paracetamol-related discharges, but not for antidepressants or for opioids. Overdose was more common in females, except for opioids. Discharges related to opioids increased in an exponential manner over the decade, five-fold in women and six-fold in men in 10 years. DISCUSSION: Increases in opioid-related presentations are of major concern. Changes in paracetamol pack-size have been associated with reduced discharge rates. In Scotland the age group with the highest rate of discharge (15-24 years) with paracetamol overdose is not the one with the highest mortality. PMID- 12589011 TI - The head-up tilt test with haemodynamic instability score in diagnosing chronic fatigue syndrome. AB - BACKGROUND: Studying patients with chronic fatigue syndrome (CFS), we have developed a method that uses a head-up tilt test (HUTT) to estimate BP and HR instability during tilt, expressed as a 'haemodynamic instability score' (HIS). AIM: To assess HIS sensitivity and specificity in the diagnosis of CFS. DESIGN: Prospective controlled study. METHODS: Patients with CFS (n=40), non-CFS chronic fatigue (n=73), fibromyalgia (n=41), neurally mediated syncope (n=58), generalized anxiety disorder (n=28), familial Mediterranean fever (n=50), arterial hypertension (n=28), and healthy subjects (n=59) were evaluated with a standardized head-up tilt test (HUTT). The HIS was calculated from blood pressure (BP) and heart rate (HR) changes during the HUTT. RESULTS: The tilt was prematurely terminated in 22% of CFS patients when postural symptoms occurred and the HIS could not be calculated. In the remainder, the median(IQR) HIS values were: CFS +2.14(4.67), non-CFS fatigue -3.98(5.35), fibromyalgia -2.81(2.62), syncope -3.7(4.36), generalized anxiety disorder -0.21(6.05), healthy controls 2.66(3.14), FMF -5.09(6.41), hypertensives -5.35(2.74) (p<0.0001 vs. CFS in all groups, except for anxiety disorder, p=NS). The sensitivity for CFS at HIS >-0.98 cut-off was 90.3% and the overall specificity was 84.5%. DISCUSSION: There is a particular dysautonomia in CFS that differs from dysautonomia in other disorders, characterized by HIS >-0.98. The HIS can reinforce the clinician's diagnosis by providing objective criteria for the assessment of CFS, which until now, could only be subjectively inferred. PMID- 12589012 TI - Cluster-randomized, controlled trial of computer-based decision support for selecting long-term anti-thrombotic therapy after acute ischaemic stroke. AB - BACKGROUND: Identifying the appropriate long-term anti-thrombotic therapy following acute ischaemic stroke is a challenging area in which computer-based decision support may provide assistance. AIM: To evaluate the influence on prescribing practice of a computer-based decision support system (CDSS) that provided patient-specific estimates of the expected ischaemic and haemorrhagic vascular event rates under each potential anti-thrombotic therapy. DESIGN: Cluster-randomized controlled trial. METHODS: We recruited patients who presented for a first investigation of ischaemic stroke or TIA symptoms, excluding those with a poor prognosis or major contraindication to anticoagulation. After observation of routine prescribing practice (6 months) in each hospital, centres were randomized for 6 months to either control (routine practice observed) or intervention (practice observed while the CDSS provided patient-specific information). We compared, between control and intervention centres, the risk reduction (estimated by the CDSS) in ischaemic and haemorrhagic vascular events achieved by long-term anti-thrombotic therapy, and the proportions of subjects prescribed the optimal therapy identified by the CDSS. RESULTS: Sixteen hospitals recruited 1952 subjects. When the CDSS provided information, the mean relative risk reduction attained by prescribing increased by 2.7 percentage units (95%CI 0.3 to 5.7) and the odds ratio for the optimal therapy being prescribed was 1.32 (0.83 to 1.80). Some 55% (5/9) of clinicians believed the CDSS had influenced their prescribing. CONCLUSIONS: Cluster-randomized trials provide excellent frameworks for evaluating novel clinical management methods. Our CDSS was feasible to implement and acceptable to clinicians, but did not substantially influence prescribing practice for anti-thrombotic drugs after acute ischaemic stroke. PMID- 12589013 TI - Superiority and equivalence in thrombolytic drugs: an interpretation. PMID- 12589014 TI - Hypokalaemia and paralysis. AB - A patient with a severe degree of hypokalaemia (1.8 mmol/l) and paralysis was brought to the emergency department. Hypokalaemic periodic paralysis was an unlikely diagnosis, because an acid-base disorder (metabolic alkalosis) and a high rate of potassium (K(+)) excretion were present. During an imaginary consultation with Professor McCance, the combination of emphasis on principles of integrative physiology, a deductive analysis, common sense, and clinical skills led to an obvious diagnosis. Nevertheless, a surprise was in store, because renal K(+) wasting persisted for almost 2 weeks after removal of the causative agent. Possible explanations for the continued kaliuresis, as well as therapeutic strategies to avoid potential complications, were considered. This case illustrates the value of applying principles of physiology in a quantitative fashion at the bedside. PMID- 12589015 TI - Type 1 autoimmune hepatitis revealed by a dysphonia related to cricoarytenoid arthritis. PMID- 12589016 TI - The trials of alternative medicine. PMID- 12589017 TI - Familial juvenile hyperuricaemic nephropathy. PMID- 12589018 TI - Failing dogma and a sloppy code. PMID- 12589020 TI - Rnf2 (Ring1b) deficiency causes gastrulation arrest and cell cycle inhibition. AB - The highly homologous Rnf2 (Ring1b) and Ring1 (Ring1a) proteins were identified as in vivo interactors of the Polycomb Group (PcG) protein Bmi1. Functional ablation of Rnf2 results in gastrulation arrest, in contrast to relatively mild phenotypes in most other PcG gene null mutants belonging to the same functional group, among which is Ring1. Developmental defects occur in both embryonic and extraembryonic tissues during gastrulation. The early lethal phenotype is reminiscent of that of the PcG-gene knockouts Eed and Ezh2, which belong to a separate functional PcG group and PcG protein complex. This finding indicates that these biochemically distinct PcG complexes are both required during early mouse development. In contrast to the strong skeletal transformation in Ring1 hemizygous mice, hemizygocity for Rnf2 does not affect vertebral identity. However, it does aggravate the cerebellar phenotype in a Bmi1 null-mutant background. Together, these results suggest that Rnf2 or Ring1-containing PcG complexes have minimal functional redundancy in specific tissues, despite overlap in expression patterns. We show that the early developmental arrest in Rnf2-null embryos is partially bypassed by genetic inactivation of the Cdkn2a (Ink4aARF) locus. Importantly, this finding implicates Polycomb-mediated repression of the Cdkn2a locus in early murine development. PMID- 12589021 TI - Functional copper at the acetyl-CoA synthase active site. AB - The bifunctional CO dehydrogenase/acetyl-CoA synthase (CODH/ACS) plays a central role in the Wood-Ljungdahl pathway of autotrophic CO(2) fixation. A recent structure of the Moorella thermoacetica enzyme revealed that the ACS active site contains a [4Fe-4S] cluster bridged to a binuclear Cu-Ni site. Here, biochemical and x-ray absorption spectroscopic (XAS) evidence is presented that the copper ion at the M. thermoacetica ACS active site is essential. Depletion of copper correlates with reduction in ACS activity and in intensity of the "NiFeC" EPR signal without affecting either the activity or the EPR spectroscopic properties associated with CODH. In contrast, Zn content is negatively correlated with ACS activity without any apparent relationship to CODH activity. Cu is also found in the methanogenic CODH/ACS from Methanosarcina thermophila. XAS studies are consistent with a distorted Cu(I)-S(3) site in the fully active enzyme in solution. Cu extended x-ray absorption fine structure analysis indicates an average Cu-S bond length of 2.25 A and a metal neighbor at 2.65 A, consistent with the Cu-Ni distance observed in the crystal structure. XAS experiments in the presence of seleno-CoA reveal a Cu-S(3)Se environment with a 2.4-A Se-Cu bond, strongly implicating a Cu-SCoA intermediate in the mechanism of acetyl-CoA synthesis. These results indicate an essential and functional role for copper in the CODH/ACS from acetogenic and methanogenic organisms. PMID- 12589022 TI - Androgen-induced recruitment of RNA polymerase II to a nuclear receptor-p160 coactivator complex. AB - The androgen receptor, like other nuclear receptors, activates target genes by binding to hormone-responsive enhancers. Here we demonstrate that androgen induces robust recruitment of androgen receptor, members of the p160 coactivator family, and CREB-binding protein p300 specifically at the distant enhancer of prostate-specific antigen (PSA) gene. Unexpectedly, we found that RNA polymerase II (Pol II) is directly recruited to the enhancer in a hormone-dependent manner, independent of the proximal promoter, and that the isolated PSA enhancer can mediate efficient androgen induction of transcription. Inhibition of the Pol II carboxyl-terminal domain kinase activity with low concentrations of flavopiridol blocks Pol II transfer from the enhancer to the promoter and selectively abolishes PSA induction by androgen. Moreover, elevated levels of the p160 coactivator ACTR/AIB1 increase both androgen-dependent and -independent PSA expression, by facilitating Pol II recruitment to the enhancer. These results support a model in which nuclear receptors and their coactivators mediate hormone induction by serving as a staging platform for Pol II recruitment. PMID- 12589023 TI - Mitotic spindle rotation and mode of cell division in the developing telencephalon. AB - The mode of neural stem cell division in the forebrain proliferative zones profoundly influences neocortical growth by regulating the number and diversity of neurons and glia. Long-term time-lapse multiphoton microscopy of embryonic mouse cortex reveals new details of the complex three-dimensional rotation and oscillation of the mitotic spindle before stem cell division. Importantly, the duration and amplitude of spindle movement predicts and specifies the eventual mode of mitotic division. These technological advances have provided dramatic data and insights into the kinetics of neural stem cell division by elucidating the involvement of spindle rotation in selection of the cleavage plane and the mode of neural stem cell division that together determine the size of the mammalian neocortex. PMID- 12589024 TI - A Saccharomyces cerevisiae mutant with increased virulence. AB - Saccharomyces cerevisiae, bakers' yeast, is not a pathogen in healthy individuals, but is increasingly isolated from immunocompromised patients. The more frequent isolation of S. cerevisiae clinically raises a number of questions concerning the origin, survival, and virulence of this organism in human hosts. Here we compare the virulence of a human isolate, a strain isolated from decaying fruit, and a common laboratory strain in a mouse infection model. We find that the plant isolate is lethal in mice, whereas the laboratory strain is avirulent. A knockout of the SSD1 gene, which alters the composition and cell wall architecture of the yeast cell surface, causes both the clinical and plant isolates to be more virulent in the mouse model of infection. The hypervirulent ssd1 Delta/ssd1 Delta yeast strain is a more potent elicitor of proinflammatory cytokines from macrophages in vitro. Our data suggest that the increased virulence of the mutant strains is a consequence of unique surface characteristics that overstimulate the proinflammatory response. PMID- 12589025 TI - Immunotherapy for choroidal neovascularization in a laser-induced mouse model simulating exudative (wet) macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of blindness after age 55 in the industrialized world. Severe loss of central vision frequently occurs with the exudative (wet) form of AMD, as a result of the formation of a pathological choroidal neovasculature (CNV) that damages the macular region of the retina. We tested the effect of an immunotherapy procedure, which had been shown to destroy the pathological neovasculature in solid tumors, on the formation of laser-induced CNV in a mouse model simulating exudative AMD in humans. The procedure involves administering an Icon molecule that binds with high affinity and specificity to tissue factor (TF), resulting in the activation of a potent cytolytic immune response against cells expressing TF. The Icon binds selectively to TF on the vascular endothelium of a CNV in the mouse and pig models and also on the CNV of patients with exudative AMD. Here we show that the Icon dramatically reduces the frequency of CNV formation in the mouse model. After laser treatment to induce CNV formation, the mice were injected either with an adenoviral vector encoding the Icon, resulting in synthesis of the Icon by vector-infected mouse cells, or with the Icon protein. The route of injection was i.v. or intraocular. The efficacy of the Icon in preventing formation of laser induced CNV depends on binding selectively to the CNV. Because the Icon binds selectively to the CNV in exudative AMD as well as to laser-induced CNV, the Icon might also be efficacious for treating patients with exudative AMD. PMID- 12589026 TI - Ends-out, or replacement, gene targeting in Drosophila. AB - Ends-in and ends-out refer to the two arrangements of donor DNA that can be used for gene targeting. Both have been used for targeted mutagenesis, but require donors of differing design. Ends-out targeting is more frequently used in mice and yeast because it gives a straightforward route to replace or delete a target locus. Although ends-in targeting has been successful in Drosophila, an attempt at ends-out targeting failed. To test whether ends-out targeting could be used in Drosophila, we applied two strategies for ends-out gene replacement at the endogenous yellow (y) locus in Drosophila. First, a mutant allele was rescued by replacement with an 8-kb y(+) DNA fragment at a rate of approximately 1/800 gametes. Second, a wild-type gene was disrupted by the insertion of a marker gene in exon 1 at a rate of approximately 1/380 gametes. The I-SceI endonuclease component alone is not sufficient for targeting: the FLP recombinase is also needed to generate the extrachromosomal donor. When both components are used we find that ends-out targeting can be approximately as efficient as ends-in targeting, and is likely to be generally useful for Drosophila gene targeting. PMID- 12589027 TI - Dietary restriction normalizes glucose metabolism and BDNF levels, slows disease progression, and increases survival in huntingtin mutant mice. AB - In addition to neurological deficits, Huntington's disease (HD) patients and transgenic mice expressing mutant human huntingtin exhibit reduced levels of brain-derived neurotrophic factor, hyperglycemia, and tissue wasting. We show that the progression of neuropathological (formation of huntingtin inclusions and apoptotic protease activation), behavioral (motor dysfunction), and metabolic (glucose intolerance and tissue wasting) abnormalities in huntingtin mutant mice, an animal model of HD, are retarded when the mice are maintained on a dietary restriction (DR) feeding regimen resulting in an extension of their life span. DR increases levels of brain-derived neurotrophic factor and the protein chaperone heat-shock protein-70 in the striatum and cortex, which are depleted in HD mice fed a normal diet. The suppression of the pathogenic processes by DR in HD mice suggests that mutant huntingtin promotes neuronal degeneration by impairing cellular stress resistance, and that the body wasting in HD is driven by the neurodegenerative process. Our findings suggest a dietary intervention that may suppress the disease process and increase the life span of humans that carry the mutant huntingtin gene. PMID- 12589028 TI - Activation of presynaptic NMDA receptors coupled to NaV1.8-resistant sodium channel C-fibers causes retrograde mechanical nociceptor sensitization. AB - The present study investigated whether activation of presynaptic N-methyl-d aspartate (NMDA) receptors in the spinal cord produces a retrograde nociceptor sensitization (hypernociception) to mechanical nonnoxious stimulus. By using an electronic version of the von Frey hair test (pressure meter), s.c. intraplantar administration of prostaglandin E(2) (PGE(2)) (50-400 ng per paw) evoked a dose related ipsilateral paw hypernociception. In contrast, intrathecal (i.t.) administration of NMDA (5-80 ng) and PGE(2) (15-150 ng) evoked dose-related bilateral paw hypernociception. The s.c. intraplantar administration of dipyrone (80-320 microg per paw) or morphine (3 and 9 microg per paw), usually used to antagonize peripheral PGE(2) (100 ng per paw), induced hypernociception and also antagonized the ipsilateral (without affecting the contralateral) paw hypernociception induced by i.t. injections of NMDA (40 ng) or PGE(2) (50 ng). These doses of drugs did not modify the basal mechanical sensitivity of control paws. This result shows that intraspinal NMDA or PGE(2) produces sensitization of the primary sensory neuron in response to mechanical stimulation. In a second series of experiments it was shown that the i.t. treatment with NaV1.8 (SNS/PN3) sodium channel antisense oligodeoxynucleotides, but not mismatch oligodeoxynucleotides, decreased the mRNA expression of sodium tetrodotoxin resistant channels on the dorsal root ganglia and abolished the mechanical hypernociception induced by i.t. administration of NMDA. Thus, our results support the suggestion that glutamate release in the spinal cord during inflammation causes retrograde hypernociception of nociceptors associated with sodium tetrodotoxin-resistant channels in primary nociceptive sensory neurons. PMID- 12589029 TI - 17AAG: low target binding affinity and potent cell activity--finding an explanation. AB - The ansamycin geldanamycin (GM) and its derivative, 17AAG, now in early clinical trials in cancer patients, have potent activity against several cancer cells at low nanomolar concentrations. The main target of these drugs is the molecular chaperone heat shock protein 90. Contrary to the high antitumor potency, the affinity of these drugs for the chaperone was determined to be approximately 1 microM. We propose that this difference can partly be explained by the physicochemical characteristics of the ansamycins. GM and 17AAG accumulate in cells, producing higher intracellular concentrations than expected. We conclude that although apparent activity for ansamycins can be seen at low nanomolar concentration, their real activity correlates with the heat shock protein 90 binding affinity and is in the low micromolar concentration range. We suggest that in the clinic, micromolar concentrations of 17AAG must accumulate in the tumor cells to achieve antitumor effects in patients comparable with ones achieved in tissue culture settings. PMID- 12589030 TI - Auditing the pharmacological accounts for Hsp90 molecular chaperone inhibitors: unfolding the relationship between pharmacokinetics and pharmacodynamics. PMID- 12589031 TI - Flavopiridol induces apoptosis in glioma cell lines independent of retinoblastoma and p53 tumor suppressor pathway alterations by a caspase-independent pathway. AB - Flavopiridol is a synthetic flavone, which inhibits growth in vitro and in vivo of several solid malignancies such as renal, prostate, and colon cancers. It is a potent cyclin-dependent kinase inhibitor presently in clinical trials. In this study, we examined the effect of flavopiridol on a panel of glioma cell lines having different genetic profiles: five of six have codeletion of p16(INK4a) and p14(ARF); three of six have p53 mutations; and one of six shows overexpression of mouse double minute-2 (MDM2) protein. Independent of retinoblastoma and p53 tumor suppressor pathway alterations, flavopiridol induced apoptosis in all cell lines but through a caspase-independent mechanism. No cleavage products for caspase 3 or its substrate poly(ADP-ribose) polymerase or caspase 8 were detected. The pan caspase inhibitor Z-VAD-fmk did not inhibit flavopiridol-induced apoptosis. Mitochondrial damage measured by cytochrome c release and transmission electron microscopy was not observed in drug-treated glioma cells. In contrast, flavopiridol treatment induced translocation of apoptosis-inducing factor from the mitochondria to the nucleus. The proteins cyclin D(1) and MDM2 involved in the regulation of retinoblastoma and p53 activity, respectively, were down regulated early after flavopiridol treatment. Given that MDM2 protein can confer oncogenic properties under certain circumstances, loss of MDM2 expression in tumor cells could promote increased chemosensitivity. After drug treatment, a low Bcl-2/Bax ratio was observed, a condition that may favor apoptosis. Taken together, the data indicate that flavopiridol has activity against glioma cell lines in vitro and should be considered for clinical development in the treatment of glioblastoma multiforme. PMID- 12589032 TI - Gene expression profiling of multiple histone deacetylase (HDAC) inhibitors: defining a common gene set produced by HDAC inhibition in T24 and MDA carcinoma cell lines. AB - Acetylation of histones in chromatin is one mechanism involved in the regulation of gene transcription and is tightly controlled by the balance of acetyltransferase and deacetylase (HDAC) activities. In cancer, some genes are repressed by the inappropriate recruitment of HDACs, e.g., tumor suppressor genes. To understand the genomic effects of HDAC inhibition on gene transcription we studied the gene expression profiles of T24 bladder and MDA breast carcinoma cells treated with three HDAC inhibitors, suberoylanilide hydroxamic acid, trichostatin A, and MS-27-275. The gene expression profiles of the HDAC inhibitors were generally similar to one another and differed substantially from those produced by structurally related inactive analogues; consequently, the changes in gene expression are mechanism-based. Hierarchical clustering of expression profiles demonstrated a greater similarity between the two hydroxamate containing inhibitors (suberoylanilide hydroxamic acid and trichostatin A) than with MS-27-275. This difference was also supported by cell phenotypic experiments. As many genes were down-regulated as up-regulated by HDAC inhibitor treatment. Comparison of the data sets defined a common ("core") set of 13 genes regulated by all of the HDAC inhibitors in three cell lines, 8 up-regulated and 5 down-regulated. Ten of 13 genes were confirmed in dose response studies in T24 cells by quantitative-PCR. The core regulated genes are involved predominantly in cell cycle/apoptosis and DNA synthesis in response to HDAC inhibitors. These data will aide in understanding the complex set of events in cells in response to chromatin remodeling induced by HDAC inhibition, which may be responsible for antitumor effects. PMID- 12589033 TI - p53-independent response to cisplatin and oxaliplatin in MMTV-ras mouse salivary tumors. AB - A transgenic mouse tumor model was used to investigate the role of p53 in tumor response to two different platinum-based chemotherapeutic agents: (a) cisplatin and (b) oxaliplatin, a diaminocyclohexane platine recently introduced into the clinic. MMTV-v-Ha-ras transgenic mice were interbred to p53-deficient mice to generate mice that develop salivary tumors either possessing or lacking p53. Tumor-bearing mice were then treated on either a 9-day schedule to assess overall tumor growth response or on a short-term treatment schedule to assess effects on cell cycle parameters and apoptosis. Both agents induced significant apoptosis and promoted overall tumor regression, regardless of the p53 status of the tumor. This is in contrast to previous studies using this model in which treatment with paclitaxel or doxorubicin promoted tumor growth arrest but not apoptosis. These findings indicate that even in the context of an activated ras gene that potentially mediates suppression of apoptosis, both cisplatin and oxaliplatin are capable of promoting an efficient p53-independent tumor response. PMID- 12589034 TI - BCR/ABL-mediated increased expression of multiple known and novel genes that may contribute to the pathogenesis of chronic myelogenous leukemia. AB - The BCR/ABL chimeric protein plays a central role in the pathogenesis of chronic myelogenous leukemia (CML). Intensive research has elucidated many signal transduction pathways activated by BCR/ABL. However, few studies addressed BCR/ABL-dependent alterations in gene expression that may contribute to the pathobiology of CML. To additionally define such downstream genes, we performed a subtractive hybridization between cord blood (CB) CD34(+) cells transduced with an MSCV-retrovirus vector containing either enhanced green fluorescent protein (eGFP) alone or p210(BCR/ABL)-internal ribosome entry site-eGFP. Thirty-four subtracted clones expressed in p210-eGFP but not eGFP-transduced CD34(+) cells have been confirmed by Northern blot and sequenced. Fifty-nine percent represent novel proteins, and 41% are homologous to known genes. Quantitative real-time PCR analysis confirmed that 14 of 14 genes tested were also overexpressed in additional populations of p210(BCR/ABL)-transduced CB CD34(+) cells, as well as in CD34(+) cells from primary newly diagnosed CML patients versus GFP-transduced CB or samples from normal donors. Western blot analysis showed that the known sequences were also overexpressed at the protein level. Treatment of BCR/ABL(+) cells with the Abl-specific tyrosine kinase inhibitor STI571 decreased expression at the mRNA as well as protein level of some but not all of the gene products. This suggests that increased gene expression is in some cases tyrosine kinase independent. Some of the overexpressed genes are implicated in cellular processes known to be disturbed in CML, including the mitogen-activated protein kinase or the ubiquitin pathway, whereas overexpression of other genes, including RAN and NUP98, may implicate new cellular pathways involved in CML. Additional characterization of downstream genes activated by BCR/ABL may lead to important new insights in the molecular mechanisms underlying CML and identify potentially novel therapeutic targets for CML. PMID- 12589035 TI - Effects of the cyclooxygenase inhibitor, piroxicam, in combination with chemotherapy on tumor response, apoptosis, and angiogenesis in a canine model of human invasive urinary bladder cancer. AB - The objectives of this study were: (a) to determine the antitumor activity and toxicity of a cyclooxygenase inhibitor (piroxicam) combined with cisplatin chemotherapy in dogs with naturally-occurring, invasive transitional cell carcinoma (TCC) of the urinary bladder; and (b) to determine the effects of this treatment on prostaglandin E(2) concentration, tumor cell proliferation and apoptosis, and angiogenesis. Pet dogs with naturally-occurring invasive TCC underwent complete tumor staging before and after 10 weeks of piroxicam/cisplatin treatment. Prostaglandin E(2) concentrations were determined by immunoassay in snap-frozen tumor tissues. Apoptosis (terminal deoxynucleotidyl transferase mediated nick end labeling assay), proliferation (proliferating cell nuclear antigen), and microvessel density were determined in formalin-fixed tissues. Urine basic fibroblast growth factor and vascular endothelial cell growth factor concentrations were determined by immunoassay. Partial remission (> or =50% reduction in tumor volume) was noted in 6 of 12 dogs treated with piroxicam/cisplatin. Renal toxicity was dose-limiting. Apoptotic index doubled with treatment in 11 of 12 dogs but was not associated with tumor response. Proliferative index decreased in five dogs, and tumor decreased in size in three of the five dogs. Change in urine basic fibroblast growth factor and vascular endothelial cell growth factor was associated with tumor response. microvessel density was not associated with tumor response. In conclusion, piroxicam/cisplatin had antitumor activity against canine TCC, a disease that closely mimics human invasive urinary bladder cancer. Strategies to prevent renal toxicity of this protocol are needed. Induction of tumor apoptosis and reduction in angiogenic factor concentrations were observed, but additional studies are needed to further define the mechanisms of the antitumor activity of piroxicam/cisplatin. PMID- 12589036 TI - Activation of adriamycin by the pH-dependent formaldehyde-releasing prodrug hexamethylenetetramine. AB - Previous studies have shown that Adriamycin can react with formaldehyde to yield an activated form of Adriamycin that can further react with DNA to yield Adriamycin-DNA adducts. Because hexamethylenetetramine (HMTA) is known to hydrolyze under cellular conditions and release six molecules of formaldehyde in a pH-dependent manner, we examined this clinical agent for its potential as a formaldehyde-releasing prodrug for the activation of Adriamycin. In IMR-32 neuroblastoma cells in culture, increasing levels of HMTA resulted in enhanced levels of Adriamycin-DNA adducts. These adducts were formed in a pH-dependent manner, with 4-fold more detected at pH 6.5 compared with pH 7.4, consistent with the known acid lability of HMTA. The resulting drug-DNA lesion was shown to be cytotoxic, with combined Adriamycin and prodrug treatment resulting in a 3-fold lower IC(50) value compared with that of Adriamycin alone. Given the acidic nature of solid tumors and the preferential release of formaldehyde from HMTA in acidic environments, HMTA therefore has some potential for localized activation of Adriamycin in solid tumors. PMID- 12589037 TI - Relevance network between chemosensitivity and transcriptome in human hepatoma cells. AB - Generally, hepatoma is not a chemosensitive tumor, and the mechanism of resistance to anticancer drugs is not fully elucidated. We aimed to comprehensively evaluate the relationship between chemosensitivity and gene expression profile in human hepatoma cells, by using microarray analysis, and analyze the data by constructing relevance networks. In eight hepatoma cell lines (HLE, HLF, Huh7, Hep3B, PLC/PRF/5, SK-Hep1, Huh6, and HepG2), the baseline expression levels of 2300 genes were measured by cDNA microarray. The concentrations of eight anticancer drugs (nimustine, mitomycin C, cisplatin, carboplatin, doxorubicin, epirubicin, mitoxantrone, and 5-fluorouracil) needed for 50% growth inhibition were examined and used as a measure of chemosensitivity. These data were combined and comprehensive pair-wise correlations between gene expression levels and the 50% growth inhibition values were calculated. Significant correlations with significance were used to construct networks of similarity. Fifty-two relations, including 42 genes, were selected. Among them, nearly 20% were various types of transporters, and most of them negatively correlated with chemosensitivity. Transporter associated with antigen processing 1 was associated with resistance to mitoxantrone, consistent with previous reports. Other transporters were not reported previously to associate with chemosensitivity. Resistance to doxorubicin and its analogue, epirubicin, were positively correlated with topoisomerase II beta expression, whereas it negatively correlated with expression of carboxypeptidases A3 and Z. Response to nimustine was associated with expression of superoxide dismutase 2. Relevance networks identified several negative correlations between gene expression and resistance, which were missed by hierarchical clustering. Our results suggested the necessity of systematically evaluating the transporting systems that may play a major role in resistance in hepatoma. This may provide useful information to modify anticancer drug action in hepatoma. PMID- 12589038 TI - Integrin-mediated tyrosine phosphorylation of Shc in T cells is regulated by protein kinase C-dependent phosphorylations of Lck. AB - Integrin receptor signals are costimulatory for mitogenesis with the T-cell receptor during T-cell activation. A subset of integrin receptors can link to the adapter protein Shc and provide a mitogenic stimulus. Using a combination of genetic and pharmacological approaches, we show herein that integrin signaling to Shc in T cells requires the receptor tyrosine phosphatase CD45, the Src family kinase member Lck, and protein kinase C. Our results suggest a model in which integrin-dependent serine phosphorylation of Lck is the critical step that determines the efficiency of Shc tyrosine phosphorylation in T cells. Serine phosphorylation of Lck is dependent on PKC and is also linked to CD45 dephosphorylation. Mutants of Lck that cannot be phosphorylated on the critical serine residues do not signal efficiently to Shc and have greatly reduced kinase activity. This signaling from integrins to Lck may be an important step in the costimulation with the T-cell receptor during lymphocyte activation. PMID- 12589039 TI - The AtC-VPS protein complex is localized to the tonoplast and the prevacuolar compartment in arabidopsis. AB - Plant cells contain several types of vacuoles with specialized functions. Although the biogenesis of these organelles is well understood at the morphological level, the machinery involved in plant vacuole formation is largely unknown. We have recently identified an Arabidopsis mutant, vcl1, that is deficient in vacuolar formation. VCL1 is homologous to a protein that regulates membrane fusion at the tonoplast in yeast. On the basis of these observations, VCL1 is predicted to play a direct role in vacuolar biogenesis and vesicular trafficking to the vacuole in plants. In this work, we show that VCL1 forms a complex with AtVPS11 and AtVPS33 in vivo. These two proteins are homologues of proteins that have a well-characterized role in membrane fusion at the tonoplast in yeast. VCL1, AtVPS11, and AtVPS33 are membrane-associated and cofractionate with tonoplast and denser endomembrane markers in subcellular fractionation experiments. Consistent with this, VCL1, AtVPS11, and AtVPS33 are found on the tonoplast and the prevacuolar compartment (PVC) by immunoelectron microscopy. We also show that a VCL1-containing complex includes SYP2-type syntaxins and is most likely involved in membrane fusion on both the PVC and tonoplast in vivo. VCL1, AtVPS11, and AtVPS33 are the first components of the vacuolar biogenesis machinery to be identified in plants. PMID- 12589040 TI - Synthetic lethal interaction of the mitochondrial phosphatidylethanolamine biosynthetic machinery with the prohibitin complex of Saccharomyces cerevisiae. AB - The majority of mitochondrial phosphatidylethanolamine (PtdEtn), a phospholipid essential for aerobic growth of yeast cells, is synthesized by phosphatidylserine decarboxylase 1 (Psd1p) in the inner mitochondrial membrane (IMM). To identify components that become essential when the level of mitochondrial PtdEtn is decreased, we screened for mutants that are synthetically lethal with a temperature-sensitive (ts) allele of PSD1. This screen unveiled mutations in PHB1 and PHB2 encoding the two subunits of the prohibitin complex, which is located to the IMM and required for the stability of mitochondrially encoded proteins. Deletion of PHB1 and PHB2 resulted in an increase of mitochondrial PtdEtn at 30 degrees C. On glucose media, phb1Delta psd1Delta and phb2Delta psd1Delta double mutants were rescued only for a limited number of generations by exogenous ethanolamine, indicating that a decrease of the PtdEtn level is detrimental for prohibitin mutants. Similar to phb mutants, deletion of PSD1 destabilizes polypeptides encoded by the mitochondrial genome. In a phb1Delta phb2Delta psd1(ts) strain the destabilizing effect is dramatically enhanced. In addition, the mitochondrial genome is lost in this triple mutant, and nuclear-encoded proteins of the IMM are assembled at a very low rate. At the nonpermissive temperature mitochondria of phb1Delta phb2Delta psd1(ts) were fragmented and aggregated. In conclusion, destabilizing effects triggered by low levels of mitochondrial PtdEtn seem to account for synthetic lethality of psd1Delta with phb mutants. PMID- 12589041 TI - Dendritic fibroblasts in three-dimensional collagen matrices. AB - Cell motility determines form and function of multicellular organisms. Most studies on fibroblast motility have been carried out using cells on the surfaces of culture dishes. In situ, however, the environment for fibroblasts is the three dimensional extracellular matrix. In the current research, we studied the morphology and motility of human fibroblasts embedded in floating collagen matrices at a cell density below that required for global matrix remodeling (i.e., contraction). Under these conditions, cells were observed to project and retract a dendritic network of extensions. These extensions contained microtubule cores with actin concentrated at the tips resembling growth cones. Platelet derived growth factor promoted formation of the network; lysophosphatidic acid stimulated its retraction in a Rho and Rho kinase-dependent manner. The dendritic network also supported metabolic coupling between cells. We suggest that the dendritic network provides a mechanism by which fibroblasts explore and become interconnected to each other in three-dimensional space. PMID- 12589042 TI - Actin filament polymerization regulates gliding motility by apicomplexan parasites. AB - Host cell entry by Toxoplasma gondii depends critically on actin filaments in the parasite, yet paradoxically, its actin is almost exclusively monomeric. In contrast to the absence of stable filaments in conventional samples, rapid-freeze electron microscopy revealed that actin filaments were formed beneath the plasma membrane of gliding parasites. To investigate the role of actin filaments in motility, we treated parasites with the filament-stabilizing drug jasplakinolide (JAS) and monitored the distribution of actin in live and fixed cells using yellow fluorescent protein (YFP)-actin. JAS treatment caused YFP-actin to redistribute to the apical and posterior ends, where filaments formed a spiral pattern subtending the plasma membrane. Although previous studies have suggested that JAS induces rigor, videomicroscopy demonstrated that JAS treatment increased the rate of parasite gliding by approximately threefold, indicating that filaments are rate limiting for motility. However, JAS also frequently reversed the normal direction of motility, disrupting forward migration and cell entry. Consistent with this alteration, subcortical filaments in JAS-treated parasites occurred in tangled plaques as opposed to the straight, roughly parallel orientation observed in control cells. These studies reveal that precisely controlled polymerization of actin filaments imparts the correct timing, duration, and directionality of gliding motility in the Apicomplexa. PMID- 12589043 TI - Podosomes display actin turnover and dynamic self-organization in osteoclasts expressing actin-green fluorescent protein. AB - Podosomes, small actin-based adhesion structures, differ from focal adhesions in two aspects: their core structure and their ability to organize into large patterns in osteoclasts. To address the mechanisms underlying these features, we imaged live preosteoclasts expressing green fluorescent protein-actin during their differentiation. We observe that podosomes always form inside or close to podosome groups, which are surrounded by an actin cloud. Fluorescence recovery after photobleaching shows that actin turns over in individual podosomes in contrast to cortactin, suggesting a continuous actin polymerization in the podosome core. The observation of podosome assemblies during osteoclast differentiation reveals that they evolve from simple clusters into rings that expand by the continuous formation of new podosomes at their outer ridge and inhibition of podosome formation inside the rings. This self-organization of podosomes into dynamic rings is the mechanism that drives podosomes at the periphery of the cell in large circular patterns. We also show that an additional step of differentiation, requiring microtubule integrity, stabilizes the podosome circles at the cell periphery to form the characteristic podosome belt pattern of mature osteoclasts. These results therefore provide a mechanism for the patterning of podosomes in osteoclasts and reveal a turnover of actin inside the podosome. PMID- 12589044 TI - Convergence of non-clathrin- and clathrin-derived endosomes involves Arf6 inactivation and changes in phosphoinositides. AB - The trafficking of two plasma membrane (PM) proteins that lack clathrin internalization sequences, major histocompatibility complex class I (MHCI), and interleukin 2 receptor alpha subunit (Tac) was compared with that of PM proteins internalized via clathrin. MHCI and Tac were internalized into endosomes that were distinct from those containing clathrin cargo. At later times, a fraction of these internalized membranes were observed in Arf6-associated, tubular recycling endosomes whereas another fraction acquired early endosomal autoantigen 1 (EEA1) before fusion with the "classical" early endosomes containing the clathrin dependent cargo, LDL. After convergence, cargo molecules from both pathways eventually arrived, in a Rab7-dependent manner, at late endosomes and were degraded. Expression of a constitutively active mutant of Arf6, Q67L, caused MHCI and Tac to accumulate in enlarged PIP(2)-enriched vacuoles, devoid of EEA1 and inhibited their fusion with clathrin cargo-containing endosomes and hence blocked degradation. By contrast, trafficking and degradation of clathrin-cargo was not affected. A similar block in transport of MHCI and Tac was reversibly induced by a PI3-kinase inhibitor, implying that inactivation of Arf6 and acquisition of PI3P are required for convergence of endosomes arising from these two pathways. PMID- 12589045 TI - Visualizing lipid raft dynamics and early signaling events during antigen receptor-mediated B-lymphocyte activation. AB - Recent biochemical evidence indicates that an early event in signal transduction by the B-cell antigen receptor (BCR) is its translocation to specialized membrane subdomains known as lipid rafts. We have taken a microscopic approach to image lipid rafts and early events associated with BCR signal transduction. Lipid rafts were visualized on primary splenic B lymphocytes from wild-type or anti-hen egg lysozyme BCR transgenic mice, and on a mature mouse B-cell line Bal 17 by using fluorescent conjugates of cholera toxin B subunit or a Lyn-based chimeric protein, which targets green fluorescent protein to the lipid raft compartment. Time-lapse imaging of B cells stimulated via the BCR with the antigen hen egg lysozyme, or surrogate for antigen anti-IgM, demonstrated that lipid rafts are highly dynamic entities, which move laterally on the surface of these cells and coalesce into large regions. These regions of aggregated lipid rafts colocalized with the BCR and tyrosine-phosphorylated proteins. Microscopic imaging of live B cells also revealed an inducible colocalization of lipid rafts with the tyrosine kinase Syk and the receptor tyrosine phosphatase CD45. These two proteins play indispensable roles in BCR-mediated signaling but are not detectable in biochemically purified lipid raft fractions. Strikingly, BCR stimulation also induced the formation of long, thread-like filopodial projections, similar to previously described structures called cytonemes. These B-cell cytonemes are rich in lipid rafts and actin filaments, suggesting that they might play a role in long-range communication and/or transportation of signaling molecules during an immune response. These results provide a window into the morphological and molecular organization of the B-cell membrane during the early phase of BCR signaling. PMID- 12589046 TI - Myosin motors and not actin comets are mediators of the actin-based Golgi-to endoplasmic reticulum protein transport. AB - We have previously reported that actin filaments are involved in protein transport from the Golgi complex to the endoplasmic reticulum. Herein, we examined whether myosin motors or actin comets mediate this transport. To address this issue we have used, on one hand, a combination of specific inhibitors such as 2,3-butanedione monoxime (BDM) and 1-[5-isoquinoline sulfonyl]-2-methyl piperazine (ML7), which inhibit myosin and the phosphorylation of myosin II by the myosin light chain kinase, respectively; and a mutant of the nonmuscle myosin II regulatory light chain, which cannot be phosphorylated (MRLC2(AA)). On the other hand, actin comet tails were induced by the overexpression of phosphatidylinositol phosphate 5-kinase. Cells treated with BDM/ML7 or those that express the MRLC2(AA) mutant revealed a significant reduction in the brefeldin A (BFA)-induced fusion of Golgi enzymes with the endoplasmic reticulum (ER). This delay was not caused by an alteration in the formation of the BFA-induced tubules from the Golgi complex. In addition, the Shiga toxin fragment B transport from the Golgi complex to the ER was also altered. This impairment in the retrograde protein transport was not due to depletion of intracellular calcium stores or to the activation of Rho kinase. Neither the reassembly of the Golgi complex after BFA removal nor VSV-G transport from ER to the Golgi was altered in cells treated with BDM/ML7 or expressing MRLC2(AA). Finally, transport carriers containing Shiga toxin did not move into the cytosol at the tips of comet tails of polymerizing actin. Collectively, the results indicate that 1) myosin motors move to transport carriers from the Golgi complex to the ER along actin filaments; 2) nonmuscle myosin II mediates in this process; and 3) actin comets are not involved in retrograde transport. PMID- 12589047 TI - Chl4p and iml3p are two new members of the budding yeast outer kinetochore. AB - Kinetochore proteins contribute to the fidelity of chromosome transmission by mediating the attachment of a specialized chromosomal region, the centromere, to the mitotic spindle during mitosis. In budding yeast, a subset of kinetochore proteins, referred to as the outer kinetochore, provides a link between centromere DNA-binding proteins of the inner kinetochore and microtubule-binding proteins. Using a combination of chromatin immunoprecipitation, in vivo localization, and protein coimmunoprecipitation, we have established that yeast Chl4p and Iml3p are outer kinetochore proteins that localize to the kinetochore in a Ctf19p-dependent manner. Chl4p interacts with the outer kinetochore proteins Ctf19p and Ctf3p, and Iml3p interacts with Chl4p and Ctf19p. In addition, Chl4p is required for the Ctf19p-Ctf3p and Ctf19p-Iml3p interactions, indicating that Chl4p is an important structural component of the outer kinetochore. These physical interaction dependencies provide insights into the molecular architecture and centromere DNA loading requirements of the outer kinetochore complex. PMID- 12589048 TI - Chemical genetic analysis of Apg1 reveals a non-kinase role in the induction of autophagy. AB - Macroautophagy is a catabolic membrane trafficking phenomenon that is observed in all eukaryotic cells in response to various stimuli, such as nitrogen starvation and challenge with specific hormones. In the yeast Saccharomyces cerevisiae, the induction of autophagy involves a direct signal transduction mechanism that affects membrane dynamics. In this system, the induction process modifies a constitutive trafficking pathway called the cytoplasm-to-vacuole targeting (Cvt) pathway, which transports the vacuolar hydrolase aminopeptidase I, from the formation of small Cvt vesicles to the formation of autophagosomes. Apg1 is one of the proteins required for the direct signal transduction cascade that modifies membrane dynamics. Although Apg1 is required for both the Cvt pathway and autophagy, we find that Apg1 kinase activity is required only for Cvt trafficking of aminopeptidase I but not for import via autophagy. In addition, the data support a novel role for Apg1 in nucleation of autophagosomes that is distinct from its catalytic kinase activity and imply a qualitative difference in the mechanism of autophagosome and Cvt vesicle formation. PMID- 12589049 TI - IkappaBalpha and p65 regulate the cytoplasmic shuttling of nuclear corepressors: cross-talk between Notch and NFkappaB pathways. AB - Notch and NFkappaB pathways are key regulators of numerous cellular events such as proliferation, differentiation, or apoptosis. In both pathways, association of effector proteins with nuclear corepressors is responsible for their negative regulation. We have previously described that expression of a p65-NFkappaB mutant that lacks the transactivation domain (p65DeltaTA) induces cytoplasmic translocation of N-CoR leading to a positive regulation of different promoters. Now, we show that cytoplasmic sequestration of p65 by IkappaBalpha is sufficient to both translocate nuclear corepressors SMRT/N-CoR to the cytoplasm and upregulate transcription of Notch-dependent genes. Moreover, p65 and IkappaBalpha are able to directly bind SMRT, and this interaction can be inhibited in a dose dependent manner by the CREB binding protein (CBP) coactivator and after TNF alpha treatment, suggesting that p65 acetylation is modulating this interaction. In agreement with this, TNF-alpha treatment results in downregulation of the Hes1 gene. Finally, we present evidence on how this mechanism may influence cell differentiation in the 32D myeloid progenitor system. PMID- 12589050 TI - Reorganization of actin cytoskeleton by the phosphoinositide metabolite glycerophosphoinositol 4-phosphate. AB - Glycerophosphoinositol 4-phosphate (GroPIns-4P) is a biologically active, water soluble phospholipase A metabolite derived from phosphatidylinositol 4-phosphate, whose cellular concentrations have been reported to increase in Ras-transformed cells. It is therefore important to understand its biological activities. Herein, we have examined whether GroPIns-4P can regulate the organization of the actin cytoskeleton, because this could be a Ras-related function involved in cell motility and metastatic invasion. We find that in serum-starved Swiss 3T3 cells, exogenously added GroPIns-4P rapidly and potently induces the formation of membrane ruffles, and, later, the formation of stress fibers. These actin structures can be regulated by the small GTPases Cdc42, Rac, and Rho. To analyze the mechanism of action of GroPIns-4P, we selectively inactivated each of these GTPases. GroPIns-4P requires active Rac and Rho, but not Cdc42, for ruffle and stress fiber formation, respectively. Moreover, GroPIns-4P induces a rapid translocation of the green fluorescent protein-tagged Rac into ruffles, and increases the fraction of GTP-bound Rac, in intact cells. The activation of Rac by GroPIns-4P was near maximal and long-lasting. Interestingly, this feature seems to be critical in the induction of actin ruffles by GroPIns-4P. PMID- 12589051 TI - Adaptor and clathrin exchange at the plasma membrane and trans-Golgi network. AB - We previously demonstrated, using fluorescence recovery after photobleaching, that clathrin in clathrin-coated pits at the plasma membrane exchanges with free clathrin in the cytosol, suggesting that clathrin-coated pits are dynamic structures. We now investigated whether clathrin at the trans-Golgi network as well as the clathrin adaptors AP2 and AP1 in clathrin-coated pits at the plasma membrane and trans-Golgi network, respectively, also exchange with free proteins in the cytosol. We found that when the budding of clathrin-coated vesicle is blocked without significantly affecting the structure of clathrin-coated pits, both clathrin and AP2 at the plasma membrane and clathrin and AP1 at the trans Golgi network exchange rapidly with free proteins in the cytosol. In contrast, when budding of clathrin-coated vesicles was blocked at the plasma membrane or trans-Golgi network by hypertonic sucrose or K(+) depletion, conditions that markedly affect the structure of clathrin-coated pits, clathrin exchange was blocked but AP2 at the plasma membrane and both AP1 and the GGA1 adaptor at the trans-Golgi network continue to rapidly exchange. We conclude that clathrin coated pits are dynamic structures with rapid exchange of both clathrin and adaptors and that adaptors are able to exchange independently of clathrin when clathrin exchange is blocked. PMID- 12589052 TI - Transforming growth factor-beta1 (TGF-beta)-induced apoptosis of prostate cancer cells involves Smad7-dependent activation of p38 by TGF-beta-activated kinase 1 and mitogen-activated protein kinase kinase 3. AB - The inhibitory Smad7, a direct target gene for transforming growth factor-beta (TGF-beta), mediates TGF-beta1-induced apoptosis in several cell types. Herein, we report that apoptosis of human prostate cancer PC-3U cells induced by TGF beta1 or Smad7 overexpression is caused by a specific activation of the p38 mitogen-activated protein kinase pathway in a TGF-beta-activated kinase 1 (TAK1)- and mitogen-activated protein kinase kinase 3 (MKK3)-dependent manner. Expression of dominant negative p38, dominant negative MKK3, or incubation with the p38 selective inhibitor [4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4 pyridyl)1H-imidazole], prevented TGF-beta1-induced apoptosis. The expression of Smad7 was required for TGF-beta-induced activation of MKK3 and p38 kinases, and endogenous Smad7 was found to interact with phosphorylated p38 in a ligand dependent manner. Ectopic expression of wild-type TAK1 promoted TGF-beta1-induced phosphorylation of p38 and apoptosis, whereas dominant negative TAK1 reduced TGF beta1-induced phosphorylation of p38 and apoptosis. Endogenous Smad7 was found to interact with TAK1, and TAK1, MKK3, and p38 were coimmunoprecipitated with Smad7 in transiently transfected COS1 cells. Moreover, ectopically expressed Smad7 enhanced the coimmunoprecipitation of HA-MKK3 and Flag-p38, supporting the notion that Smad7 may act as a scaffolding protein and facilitate TAK1- and MKK3 mediated activation of p38. PMID- 12589053 TI - Differential roles of Smad1 and p38 kinase in regulation of peroxisome proliferator-activating receptor gamma during bone morphogenetic protein 2 induced adipogenesis. AB - Bone morphogenetic protein 2 (BMP2) promotes the differentiation of undifferentiated mesenchymal cells into adipocytes. To investigate the molecular mechanisms that regulate this differentiation process, we studied the relationship between BMP2 signaling and peroxisome proliferator-activating receptor gamma (PPARgamma) during adipogenesis of mesenchymal cells by using pluripotent mesenchymal cell line C3H10T1/2. In C3H10T1/2 cells, BMP2 induced expression of PPARgamma along with adipogenesis. Overexpression of Smad6, a natural antagonist for Smad1, blocked PPARgamma expression and adipocytic differentiation induced by BMP2. Overexpression of dominant-negative PPARgamma also diminished adipocytic differentiation of C3H10T1/2 cells, suggesting the central role of PPARgamma in BMP2-induced adipocytic differentiation. Specific inhibitors for p38 kinase inhibited BMP2-induced adipocytic differentiation and transcriptional activation of PPARgamma, whereas overexpression of Smad6 had no effect on transcriptional activity of PPARgamma. Furthermore, activation of p38 kinase by overexpression of TAK1 and TAB1, without affecting PPARgamma expression, led the up-regulation of transcriptional activity of PPARgamma. These results suggest that both Smad and p38 kinase signaling are concomitantly activated and responsible for BMP2-induced adipocytic differentiation by inducing and up-regulating PPARgamma, respectively. Thus, BMP2 controls adipocytic differentiation by using two distinct signaling pathways that play differential roles in this process in C3H10T1/2 cells. PMID- 12589054 TI - Telomeric protein distributions and remodeling through the cell cycle in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, telomeric DNA is protected by a nonnucleosomal protein complex, tethered by the protein Rap1. Rif and Sir proteins, which interact with Rap1p, are thought to have further interactions with conventional nucleosomic chromatin to create a repressive structure that protects the chromosome end. We showed by microarray analysis that Rif1p association with the chromosome ends extends to subtelomeric regions many kilobases internal to the terminal telomeric repeats and correlates strongly with the previously determined genomic footprints of Rap1p and the Sir2-4 proteins in these regions. Although the end-protection function of telomeres is essential for genomic stability, telomeric DNA must also be copied by the conventional DNA replication machinery and replenished by telomerase, suggesting that transient remodeling of the telomeric chromatin might result in distinct protein complexes at different stages of the cell cycle. Using chromatin immunoprecipitation, we monitored the association of Rap1p, Rif1p, Rif2p, and the protein component of telomerase, Est2p, with telomeric DNA through the cell cycle. We provide evidence for dynamic remodeling of these components at telomeres. PMID- 12589055 TI - Identification of a novel "chromosome scaffold" protein that associates with Tec elements undergoing en masse elimination in Euplotes crassus. AB - During macronuclear development in the ciliate Euplotes crassus, the highly repetitive, transposon-like Tec elements possess an unusual chromatin structure. We observed that the Tec element chromatin is highly resistant to salt extraction and behaves like a nuclear matrix/chromosome scaffold-associated structure. Standard matrix/scaffold extraction procedures identified two major proteins: 1) an ~140-kDa protein that seems to be topoisomerase II based on its reactivity with anti-topoisomerase II antibodies, and 2) an 85-kDa protein that we further purified by acid extraction and have shown to be a novel protein by sequence analysis of its gene. The 85-kDa protein (p85) is a developmental stage-specific protein and is located exclusively in the developing macronucleus. Immunolocalization studies of p85 show that it colocalizes with topoisomerase II in chromatin. In addition, in situ hybridization combined with immunofluorescence localization of the proteins indicates that 100% of the Tec elements colocalize with 70% of the p85, whereas no significant colocalization with a total macronuclear sequence-specific probe is observed. p85 is the first developmental stage-specific protein identified as being specifically associated with sequences undergoing elimination in E. crassus. PMID- 12589056 TI - IKKalpha regulates mitogenic signaling through transcriptional induction of cyclin D1 via Tcf. AB - The Wnt/beta-catenin/Tcf and IkappaB/NF-kappaB cascades are independent pathways involved in cell cycle control, cellular differentiation, and inflammation. Constitutive Wnt/beta-catenin signaling occurs in certain cancers from mutation of components of the pathway and from activating growth factor receptors, including RON and MET. The resulting accumulation of cytoplasmic and nuclear beta catenin interacts with the Tcf/LEF transcription factors to induce target genes. The IkappaB kinase complex (IKK) that phosphorylates IkappaB contains IKKalpha, IKKbeta, and IKKgamma. Here we show that the cyclin D1 gene functions as a point of convergence between the Wnt/beta-catenin and IkappaB pathways in mitogenic signaling. Mitogenic induction of G(1)-S phase progression and cyclin D1 expression was PI3K dependent, and cyclin D1(-/-) cells showed reduced PI3K dependent S-phase entry. PI3K-dependent induction of cyclin D1 was blocked by inhibitors of PI3K/Akt/IkappaB/IKKalpha or beta-catenin signaling. A single Tcf site in the cyclin D1 promoter was required for induction by PI3K or IKKalpha. In IKKalpha(-/-) cells, mitogen-induced DNA synthesis, and expression of Tcf responsive genes was reduced. Reintroduction of IKKalpha restored normal mitogen induction of cyclin D1 through a Tcf site. In IKKalpha(-/-) cells, beta-catenin phosphorylation was decreased and purified IKKalpha was sufficient for phosphorylation of beta-catenin through its N-terminus in vitro. Because IKKalpha but not IKKbeta induced cyclin D1 expression through Tcf activity, these studies indicate that the relative levels of IKKalpha and IKKbeta may alter their substrate and signaling specificities to regulate mitogen-induced DNA synthesis through distinct mechanisms. PMID- 12589057 TI - Nup98 localizes to both nuclear and cytoplasmic sides of the nuclear pore and binds to two distinct nucleoporin subcomplexes. AB - The vertebrate nuclear pore is an enormous structure that spans the double membrane of the nuclear envelope. In yeast, most nucleoporins are found symmetrically on both the nuclear and cytoplasmic sides of the structure. However, in vertebrates most nucleoporins have been localized exclusively to one side of the nuclear pore. Herein, we show, by immunofluorescence and immunoelectron microscopy, that Nup98 is found on both sides of the pore complex. Additionally, we find that the pore-targeting domain of Nup98 interacts directly with the cytoplasmic nucleoporin Nup88, a component of the Nup214, Nup88, Nup62 subcomplex. Nup98 was previously described to interact with the nuclear-oriented Nup160, 133, 107, 96 complex through direct binding to Nup96. Interestingly, the same site within Nup98 is involved in binding to both Nup88 and Nup96. Autoproteolytic cleavage of the Nup98 C terminus is required for both of these binding interactions. When cleavage is blocked by a point mutation, a minimal eight amino acids downstream of the cleavage site is sufficient to prevent most binding to either Nup96 or Nup88. Thus, Nup98 interacts with both faces of the nuclear pore, a localization in keeping with its previously described nucleocytoplasmic shuttling activity. PMID- 12589058 TI - Raft-mediated trafficking of apical resident proteins occurs in both direct and transcytotic pathways in polarized hepatic cells: role of distinct lipid microdomains. AB - In polarized hepatic cells, pathways and molecular principles mediating the flow of resident apical bile canalicular proteins have not yet been resolved. Herein, we have investigated apical trafficking of a glycosylphosphatidylinositol-linked and two single transmembrane domain proteins on the one hand, and two polytopic proteins on the other in polarized HepG2 cells. We demonstrate that the former arrive at the bile canalicular membrane via the indirect transcytotic pathway, whereas the polytopic proteins reach the apical membrane directly, after Golgi exit. Most importantly, cholesterol-based lipid microdomains ("rafts") are operating in either pathway, and protein sorting into such domains occurs in the biosynthetic pathway, largely in the Golgi. Interestingly, rafts involved in the direct pathway are Lubrol WX insoluble but Triton X-100 soluble, whereas rafts in the indirect pathway are both Lubrol WX and Triton X-100 insoluble. Moreover, whereas cholesterol depletion alters raft-detergent insolubility in the indirect pathway without affecting apical sorting, protein missorting occurs in the direct pathway without affecting raft insolubility. The data implicate cholesterol as a traffic direction-determining parameter in the direct apical pathway. Furthermore, raft-cargo likely distinguishing single vs. multispanning membrane anchors, rather than rafts per se (co)determine the sorting pathway. PMID- 12589059 TI - EpsinR: an ENTH domain-containing protein that interacts with AP-1. AB - We have used GST pulldowns from A431 cell cytosol to identify three new binding partners for the gamma-adaptin appendage: Snx9, ARF GAP1, and a novel ENTH domain containing protein, epsinR. EpsinR is a highly conserved protein that colocalizes with AP-1 and is enriched in purified clathrin-coated vesicles. However, it does not require AP-1 to get onto membranes and remains membrane-associated in AP-1 deficient cells. Moreover, although epsinR binds AP-1 via its COOH-terminal domain, its NH(2)-terminal ENTH domain can be independently recruited onto membranes, both in vivo and in vitro. Brefeldin A causes epsinR to redistribute into the cytosol, and recruitment of the ENTH domain requires GTPgammaS, indicating that membrane association is ARF dependent. In protein-lipid overlay assays, the epsinR ENTH domain binds to PtdIns(4)P, suggesting a possible mechanism for ARF-dependent recruitment onto TGN membranes. When epsinR is depleted from cells by RNAi, cathepsin D is still correctly processed intracellularly to the mature form. This indicates that although epsinR is likely to be an important component of the AP-1 network, it is not necessary for the sorting of lysosomal enzymes. PMID- 12589060 TI - Microtubule organization requires cell cycle-dependent nucleation at dispersed cytoplasmic sites: polar and perinuclear microtubule organizing centers in the plant pathogen Ustilago maydis. AB - Growth of most eukaryotic cells requires directed transport along microtubules (MTs) that are nucleated at nuclear-associated microtubule organizing centers (MTOCs), such as the centrosome and the fungal spindle pole body (SPB). Herein, we show that the pathogenic fungus Ustilago maydis uses different MT nucleation sites to rearrange MTs during the cell cycle. In vivo observation of green fluorescent protein-MTs and MT plus-ends, tagged by a fluorescent EB1 homologue, provided evidence for antipolar MT orientation and dispersed cytoplasmic MT nucleating centers in unbudded cells. On budding gamma-tubulin containing MTOCs formed at the bud neck, and MTs reorganized with >85% of all minus-ends being focused toward the growth region. Experimentally induced lateral budding resulted in MTs that curved out of the bud, again supporting the notion that polar growth requires polar MT nucleation. Depletion or overexpression of Tub2, the gamma tubulin from U. maydis, affected MT number in interphase cells. The SPB was inactive in G2 phase but continuously recruited gamma-tubulin until it started to nucleate mitotic MTs. Taken together, our data suggest that MT reorganization in U. maydis depends on cell cycle-specific nucleation at dispersed cytoplasmic sites, at a polar MTOC and the SPB. PMID- 12589061 TI - RACK1 regulates integrin-mediated adhesion, protrusion, and chemotactic cell migration via its Src-binding site. AB - Mammalian cDNA expression cloning was used to identify novel regulators of integrin-mediated cell-substratum adhesions. Using a focal adhesion morphology screen, we identified a cDNA with homology to a receptor for activated protein kinase C (RACK1) that induced a loss of central focal adhesions and stress fibers in CHO-K1 cells. The identified cDNA was a C-terminal truncated form of RACK1 that had one of the putative protein kinase C binding sites but lacked the region proposed to bind the beta integrin cytoplasmic domain and the tyrosine kinase Src. To investigate the role of RACK1 during cell spreading and migration, we tagged RACK1, a C-terminal truncated RACK1 and a point mutant that does not bind Src (RACK Y246F) with green fluorescent protein and expressed them in CHO-K1 cells. We found that RACK1 regulates the organization of focal adhesions and that it localizes to a subset of nascent focal complexes in areas of protrusion that contain paxillin but not vinculin. We also found that RACK1 regulates cell protrusion and chemotactic migration through its Src binding site. Together, these findings suggest that RACK1 regulates adhesion, protrusion, and chemotactic migration through its interaction with Src. PMID- 12589062 TI - Involvement of the Arp2/3 complex and Scar2 in Golgi polarity in scratch wound models. AB - Cell motility and cell polarity are essential for morphogenesis, immune system function, and tissue repair. Many animal cells move by crawling, and one main driving force for movement is derived from the coordinated assembly and disassembly of actin filaments. As tissue culture cells migrate to close a scratch wound, this directional extension is accompanied by Golgi apparatus reorientation, to face the leading wound edge, giving the motile cell inherent polarity aligned relative to the wound edge and to the direction of cell migration. Cellular proteins essential for actin polymerization downstream of Rho family GTPases include the Arp2/3 complex as an actin nucleator and members of the Wiskott-Aldrich Syndrome protein (WASP) family as activators of the Arp2/3 complex. We therefore analyzed the involvement of the Arp2/3 complex and WASP family proteins in in vitro wound healing assays using NIH 3T3 fibroblasts and astrocytes. In NIH 3T3 cells, we found that actin and Arp2/3 complex contributed to cell polarity establishment. Moreover, overexpression of N-terminal fragments of Scar2 (but not N-WASP or Scar1 or Scar3) interfere with NIH 3T3 Golgi polarization but not with cell migration. In contrast, actin, Arp2/3, and WASP family proteins did not appear to be involved in Golgi polarization in astrocytes. Our results thus indicate that the requirement for Golgi polarity establishment is cell-type specific. Furthermore, in NIH 3T3 cells, Scar2 and the Arp2/3 complex appear to be involved in the establishment and maintenance of Golgi polarity during directed migration. PMID- 12589063 TI - In vitro and in vivo interactions of DNA ligase IV with a subunit of the condensin complex. AB - Several findings have revealed a likely role for DNA ligase IV, and interacting protein XRCC4, in the final steps of mammalian DNA double-strand break repair. Recent evidence suggests that the human DNA ligase IV protein plays a critical role in the maintenance of genomic stability. To identify protein-protein interactions that may shed further light on the molecular mechanisms of DSB repair and the biological roles of human DNA ligase IV, we have used the yeast two-hybrid system in conjunction with traditional biochemical methods. These efforts have resulted in the identification of a physical association between the DNA ligase IV polypeptide and the human condensin subunit known as hCAP-E. The hCAP-E polypeptide, a member of the Structural Maintenance of Chromosomes (SMC) super-family of proteins, coimmunoprecipitates from cell extracts with DNA ligase IV. Immunofluorescence studies reveal colocalization of DNA ligase IV and hCAP-E in the interphase nucleus, whereas mitotic cells display colocalization of both polypeptides on mitotic chromosomes. Strikingly, the XRCC4 protein is excluded from the area of mitotic chromosomes, suggesting the formation of specialized DNA ligase IV complexes subject to cell cycle regulation. We discuss our findings in light of known and hypothesized roles for ligase IV and the condensin complex. PMID- 12589065 TI - A truncated form of KlLsm4p and the absence of factors involved in mRNA decapping trigger apoptosis in yeast. AB - The LSM4 gene of Saccharomyces cerevisiae codes for an essential protein involved in pre-mRNA splicing and also in mRNA decapping, a crucial step for mRNA degradation. We previously demonstrated that the first 72 amino acids of the Kluyveromyces lactis Lsm4p (KlLsm4p), which contain the Sm-like domains, can restore cell viability in both K. lactis and S. cerevisiae cells not expressing the endogenous protein. However, the absence of the carboxy-terminal region resulted in a remarkable loss of viability in stationary phase cells (). Herein, we demonstrate that S. cerevisiae cells expressing the truncated LSM4 protein of K. lactis showed the phenotypic markers of yeast apoptosis such as chromatin condensation, DNA fragmentation, and accumulation of reactive oxygen species. The study of deletion mutants revealed that apoptotic markers were clearly evident also in strains lacking genes involved in mRNA decapping, such as LSM1, DCP1, and DCP2, whereas a slight effect was observed in strains lacking the genes DHH1 and PAT1. This is the first time that a connection between mRNA stability and apoptosis is reported in yeast, pointing to mRNA decapping as the crucial step responsible of the observed apoptotic phenotypes. PMID- 12589064 TI - Mammalian ykt6 is a neuronal SNARE targeted to a specialized compartment by its profilin-like amino terminal domain. AB - SNAREs are required for specific membrane fusion throughout the endomembrane system. Here we report the characterization of rat ykt6, a prenylated SNARE selectively expressed in brain neurons. Immunofluorescence microscopy in neuronal and neuroendocrine cell lines revealed that membrane-associated ykt6 did not colocalize significantly with any conventional markers of endosomes, lysosomes, or the secretory pathway. However, ykt6-containing membranes displayed very minor overlaps with lysosomes and dense-core secretory granules and were similar to lysosomes in buoyant density. Thus, ykt6 appears to be specialized for the trafficking of a unique membrane compartment, perhaps related to lysosomes, involved in aspects of neuronal function. Targeting of this SNARE to the ykt6 compartment was mediated by its profilin-like amino-terminal domain, even in the absence of protein prenylation. Although several other R-SNAREs contain related amino-terminal domains, only the ykt6 version was able to confer the specialized localization. Rat ykt6, which contains an arginine in its SNARE motif zero-layer, was found to behave like other R-SNAREs in its SNARE assembly properties. Interestingly, cytosolic ykt6, constituting more than half of the total cellular pool, appeared to be conformationally inactive for SNARE complex assembly, perhaps indicative of a regulatory mechanism that prevents promiscuous and potentially deleterious SNARE interactions. PMID- 12589066 TI - Cdc50p, a conserved endosomal membrane protein, controls polarized growth in Saccharomyces cerevisiae. AB - During the cell cycle of the yeast Saccharomyces cerevisiae, the actin cytoskeleton and the growth of cell surface are polarized, mediating bud emergence, bud growth, and cytokinesis. We identified CDC50 as a multicopy suppressor of the myo3 myo5-360 temperature-sensitive mutant, which is defective in organization of cortical actin patches. The cdc50 null mutant showed cold sensitive cell cycle arrest with a small bud as reported previously. Cortical actin patches and Myo5p, which are normally localized to polarization sites, were depolarized in the cdc50 mutant. Furthermore, actin cables disappeared, and Bni1p and Gic1p, effectors of the Cdc42p small GTPase, were mislocalized in the cdc50 mutant. As predicted by its amino acid sequence, Cdc50p appears to be a transmembrane protein because it was solubilized from the membranes by detergent treatment. Cdc50p colocalized with Vps21p in endosomal compartments and was also localized to the class E compartment in the vps27 mutant. The cdc50 mutant showed defects in a late stage of endocytosis but not in the internalization step. It showed, however, only modest defects in vacuolar protein sorting. Our results indicate that Cdc50p is a novel endosomal protein that regulates polarized cell growth. PMID- 12589067 TI - Hepatocyte growth factor switches orientation of polarity and mode of movement during morphogenesis of multicellular epithelial structures. AB - Epithelial cells form monolayers of polarized cells with apical and basolateral surfaces. Madin-Darby canine kidney epithelial cells transiently lose their apico basolateral polarity and become motile by treatment with hepatocyte growth factor (HGF), which causes the monolayer to remodel into tubules. HGF induces cells to produce basolateral extensions. Cells then migrate out of the monolayer to produce chains of cells, which go on to form tubules. Herein, we have analyzed the molecular mechanisms underlying the production of extensions and chains. We find that cells switch from an apico-basolateral polarization in the extension stage to a migratory cell polarization when in chains. Extension formation requires phosphatidyl-inositol 3-kinase activity, whereas Rho kinase controls their number and length. Microtubule dynamics and cell division are required for the formation of chains, but not for extension formation. Cells in the monolayer divide with their spindle axis parallel to the monolayer. HGF causes the spindle axis to undergo a variable "seesaw" motion, so that a daughter cells can apparently leave the monolayer to initiate a chain. Our results demonstrate the power of direct observation in investigating how individual cell behaviors, such as polarization, movement, and division are coordinated in the very complex process of producing multicellular structures. PMID- 12589068 TI - Geldanamycin treatment ameliorates the response to LPS in murine macrophages by decreasing CD14 surface expression. AB - Geldanamycin (GA) is an antibiotic produced by Actinomyces, which specifically inhibits the function of the heat shock protein 90 family. Treatment of a murine macrophage cell line (J774) with GA resulted in a reduced response to Escherichia coli lipopolysaccharide (LPS) as visualized by a decrease of NF-kappaB translocation into the nucleus and secretion of tumor necrosis factor alpha (TNF alpha). To elucidate the mechanism of this effect, the expression of CD14, the formal LPS receptor, was analyzed. Cells treated with GA showed a reduced level of surface CD14 detected by immunostaining, whereas the expression of other surface receptors, such as FC-gamma receptor and tumor necrosis factor receptors (TNF-R1 and TNF-R2), was unaffected. The reduced surface level of CD14 was not due to a reduction in its expression because CD14 steady state mRNA levels or the total cellular pool of CD14 was not altered by GA treatment. Surface CD14 was more rapidly internalized after GA treatment (2-3 h) than after incubation with cycloheximide. Immunostaining of permeabilized cells after GA treatment revealed a higher intracellular content of CD14 colocalizing with calnexin, an endoplasmic reticulum (ER) protein. These results suggest that the decrease in CD14 surface expression after GA treatment is due to rapid internalization without new replacement. These effects may be due to the inhibition of Hsp90 and Grp94 by GA in macrophages. PMID- 12589069 TI - Identification of a novel leucine-rich repeat protein as a component of flagellar radial spoke in the Ascidian Ciona intestinalis. AB - Axonemes are highly organized microtubule-based structures conserved in many eukaryotes. In an attempt to study axonemes by a proteomics approach, we selectively cloned cDNAs of axonemal proteins by immunoscreening the testis cDNA library from the ascidian Ciona intestinalis by using an antiserum against whole axonemes. We report here a 37-kDa protein of which cDNA occurred most frequently among total positive clones. This protein, named LRR37, belongs to the class of SDS22+ leucine-rich repeat (LRR) family. LRR37 is different from the LRR outer arm dynein light chain reported in Chlamydomonas and sea urchin flagella, and thus represents a novel axonemal LRR protein. Immunoelectron microscopy by using a polyclonal antibody against LRR37 showed that it is localized on the tip of the radial spoke, most likely on the spoke head. The LRR37 protein in fact seems to form a complex together with radial spoke protein 3 in a KI extract of the axonemes. These results suggest that LRR37 is a component of the radial spoke head and is involved in the interaction with other radial spoke components or proteins in the central pair projection. PMID- 12589071 TI - Direct evidence for a critical role of myosin II in budding yeast cytokinesis and the evolvability of new cytokinetic mechanisms in the absence of myosin II. AB - In the budding yeast Saccharomyces cerevisiae, an actomyosin-based contractile ring is present during cytokinesis, as occurs in animal cells. However, the precise requirement for this structure during budding yeast cytokinesis has been controversial. Here we show that deletion of MYO1, the single myosin II gene, is lethal in a commonly used strain background. The terminal phenotype of myo1Delta is interconnected chains of cells, suggestive of a cytokinesis defect. To further investigate the role of Myo1p in cytokinesis, we conditionally disrupted Myo1 function by using either a dominant negative Myo1p construct or a strain where expression of Myo1p can be shut-off. Both ways of disruption of Myo1 function result in a failure in cytokinesis. Additionally, we show that a myo1Delta strain previously reported to grow nearly as well as the wild type contains a single genetic suppressor that alleviates the severe cytokinesis defects of myo1Delta. Using fluorescence time-lapse imaging and electron microscopy techniques, we show that cytokinesis in this strain is achieved through formation of multiple aberrant septa. Taken together, these results strongly suggest that the actomyosin ring is crucial for successful cytokinesis in budding yeast, but new cytokinetic mechanisms can evolve through genetic changes when myosin II function is impaired. PMID- 12589070 TI - Pyruvate carboxylase is an essential protein in the assembly of yeast peroxisomal oligomeric alcohol oxidase. AB - Hansenula polymorpha ass3 mutants are characterized by the accumulation of inactive alcohol oxidase (AO) monomers in the cytosol, whereas other peroxisomal matrix proteins are normally activated and sorted to peroxisomes. These mutants also have a glutamate or aspartate requirement on minimal media. Cloning of the corresponding gene resulted in the isolation of the H. polymorpha PYC gene that encodes pyruvate carboxylase (HpPyc1p). HpPyc1p is a cytosolic, anapleurotic enzyme that replenishes the tricarboxylic acid cycle with oxaloacetate. The absence of this enzyme can be compensated by addition of aspartate or glutamate to the growth media. We show that HpPyc1p protein but not the enzyme activity is essential for import and assembly of AO. Similar results were obtained in the related yeast Pichia pastoris. In vitro studies revealed that HpPyc1p has affinity for FAD and is capable to physically interact with AO protein. These data suggest that in methylotrophic yeast pyruvate carboxylase plays a dual role in that, besides its well-characterized metabolic function as anapleurotic enzyme, the protein fulfils a specific role in the AO sorting and assembly process, possibly by mediating FAD-binding to AO monomers. PMID- 12589072 TI - Pex7p and Pex20p of Neurospora crassa function together in PTS2-dependent protein import into peroxisomes. AB - Recruiting matrix proteins with a peroxisomal targeting signal type 2 (PTS2) to the peroxisomal membrane requires species-specific factors. In Saccharomyces cerevisiae, the PTS2 receptor Pex7p acts in concert with the redundant Pex18p/Pex21p, whereas in Yarrowia lipolytica, Pex20p might unite the function of both S. cerevisiae peroxins. Herein, the genome of the filamentous fungus Neurospora crassa was analyzed for peroxin-encoding genes. We identified a set of 18 peroxins that resembles that of Y. lipolytica rather than that of S. cerevisiae. Interestingly, proteins homologous to both S. cerevisiae Pex7p and Y. lipolytica Pex20p exist in N. crassa. We report on the isolation of these PTS2 specific peroxins and demonstrate that NcPex20p can substitute for S. cerevisiae Pex18p/Pex21p, but not for ScPex7p. Like Pex18p, NcPex20p did not bind PTS2 protein or the docking proteins in the absence of ScPex7p. Rather, NcPex20p was required before docking to form an import-competent complex of cargo-loaded PTS2 receptors. NcPex7p did not functionally replace yeast Pex7p, probably because the N. crassa PTS2 receptor failed to associate with Pex18p/Pex21p. However, once NcPex7p and NcPex20p had been coexpressed, it proved possible to replace yeast Pex7p. Pex20p and Pex18p/Pex21p are therefore true orthologues, both of which are in need of Pex7p for PTS2 protein import. PMID- 12589073 TI - His-Asp phosphorelay signal transduction in higher plants: receptors and response regulators for cytokinin signaling in Arabidopsis thaliana. AB - Bacteria have devised phosphotransfer signaling mechanisms for eliciting a variety of adaptive responses to their environment. These mechanisms are collectively referred to as two-component regulatory systems. Each system generally consists of a sensor protein histidine kinase, which is anchored in the cell membrane, and a cytoplasmic response regulator, whose activity is modulated by the sensor. Most response regulators are transcription factors. In this review, we briefly introduce the established concept on bacterial two-component regulatory systems, using the Agrobacterium VirA-VirG system as an example, and give the evidence for the existence of quite similar systems in higher plants, such as the signal transduction induced by the phytohormone cytokinin. The Arabidopsis CRE1 histidine kinase and its related proteins AHK2 and AHK3 perceive cytokinins in the environment and transduce a signal, presumably through the AHP bridge components that carry the histidine-containing phosphotransfer (HPt) domain, to the ARR1 response regulator that transcriptionally activates genes immediately responsive to cytokinins. In addition, this signal transfer process appears to participate in cross-talk with signaling systems that respond to daylight and another phytohormone, ethylene, through an intracellular pool of several ARR1-like molecular species and the AHP components. PMID- 12589074 TI - A plant Y chromosome-STS marker encoding a degenerate retrotransposon. AB - The dioecious plant Silene latifolia has both X and Y sex chromosomes. Male specific random amplified polymorphic DNA (RAPD) fragments were analyzed to identify Y-chromosome-linked sequences. One of the RAPD fragments, MS4, was converted into a more reliable and reproducible sequence-tagged site (STS) marker. A set of MS4 STS primers was used to amplify two genomic DNA fragments (MS4a and MS4b) from a male plant and one (MS4a) from a female plant, which indicates that MS4b is located on the Y chromosome. Sequence analysis revealed that MS4a encoded a gag protein of a Ty3-gypsy-like retrotransposon. A 147-bp region from the middle of MS4a was deleted in MS4b. The MS4b sequence was not detected in the most closely related dioecious species, S. dioica. This suggests that a retrotransposon with the MS4b sequence has degenerated recently on the Y chromosome. PMID- 12589075 TI - Two new Fagopyrum (Polygonaceae) species, F. gracilipedoides and F. jinshaense from Yunnan, China. AB - Two new Fagopyrum (Polygonaceae) species, F. gracilipedoides and F. jinshaense, are described. Fagopyrum gracilipedoides was only found at Baoshan village, Lijiang district, Yunnan province, China. This species resembles the weedy species, F. gracilipes in morphology but they differ in both mating system and ploidy level; F. gracilipedoides is a heterostylous self-incompatible diploid species (2n = 16), whereas F. gracilipes is a self-compatible tetraploid species (2n = 32). Fagopyrum jinshaense was found along the Jinsha River valley near the border of Sichuan, Yunnan, and Tibet in China. Fagopyrum jinshaense is morphologically similar to F. gilesii, but they differ in the morphology of their inflorescences. Fagopyrum jinshaense has long spike-like inflorescences, whereas F. gilesii has a compact head-like one. Both F. jinshaense and F. gilesii are self-incompatible with heterostylous flowers and are diploid. Interspecific crosses between the two new species and already known species in the urophyllum group of Fagopyrum resulted in the production of self- and cross-sterile hybrids in the species combinations attempted. These results indicated that both new species are reproductively isolated from other Fagopyrum species. Molecular phylogenetic analysis based on the nucleotide sequence of the trnK gene intron in chloroplast DNA revealed the following points: 1) F. gracilipedoides, F. rubifolium and F. gracilipes / F. capillatum showed a trifurcating relationship and 2) F. jinshaense formed a clade with F. pleioramosum, F. callanthum, and F. macrocarpum, and was only distantly related to F. gilesii. In the clade of F. capillatum, F. gracilipes, F. gracilipedoides and F. rubifolium, the breakdown of self-incompatibility accompanying genome duplication seems to have occurred twice independently. PMID- 12589076 TI - Plasmon analysis of Triticum (wheat) and Aegilops. 2. Characterization and classification of 47 plasmons based on their effects on common wheat phenotype. AB - This article comprises our final remarks on the phenotypic effects of alien plasmons on common wheat. Twenty-one vegetative, reproductive, and seed characters of 551 alloplasmic lines of 12 common wheat genotypes with 46 alloplasmons, and as the control, their euplasmic lines were investigated. Effects of genotype, plasmon, and their interaction had high statistical significance for all the characters investigated, whereas phenotypic variations attributable to the alien plasmons were relatively small. Individual plasmon types are characterized by their primary effects on 21 characters. Genotype x plasmon effects on two representative characters, heading date and plant height, are described in detail. Cluster and principal component analyses of the phenotypic effects of the 47 plasmons yielded 22 groups. The relationships between these phenotype-based groups and those defined by molecular differences in organellar genomes were examined. A significant correlation was found with some explainable discrepancies. For efficient plasmon identification, use of six of the present 12 genotypes is proposed. The key for plasmon classification is provided. Our findings indicate that alien plasmons may be of limited value in future wheat breeding, but that the plasmon diversity that exists in Triticum and Aegilops species is of great significance for understanding the evolution of these genera. PMID- 12589077 TI - Genetic diversity of three Elymus species indigenous to Japan and East Asia (E. tsukushiensis, E. humidus and E. dahuricus) detected by AFLP. AB - With the aim of broadening the gene pool of Triticeae for crop improvement, the genetic diversity of three hexaploid Elymus species indigenous to Japan and East Asia (E. tsukushiensis, E. humidus and E. dahuricus) was evaluated using amplified fragment length polymorphism (AFLP) markers. A total of 34 accessions: five of E. tsukushiensis, 13 of E. humidus, 15 of E. dahuricus, and one tetraploid Elymus species as an outgroup, were analyzed. The AFLP analysis was conducted with 12 selective primer combinations. In all, 329 bands were scored. The average number of bands scored per accession was 135.3. Among the 34 accessions, no polymorphism was detected among 10 accessions of E. humidus and among three accessions of E. tsukushiensis. Each species showed a diagnostic band pattern. One accession of E. humidus showed a band pattern intermediate between those of E. humidus and E. tsukushiensis, suggesting the occurrence of interspecific introgression. The level of intraspecific variation was highest in E. dahuricus (1-psb = 0.17, pi = 0.010), followed by E. tsukushiensis (1-psb = 0.14, pi = 0.008) and E. humidus (1-psb = 0.03, pi = 0.002). The level of interspecific variation was higher than that of intraspecific variation in all the pairs compared. The level of divergence between E. humidus and E. tsukushiensis was relatively low, but even so it was two times higher than that of the intraspecific variation for each species. The phylogenetic tree constructed here showed clear separations of all the species. The results of this study indicate a clear divergence of E. humidus from E. tsukushiensis in spite of the morphological similarity between these two species. Based on the values of nucleotide diversity, the time of divergence between E. humidus and E. tsukushiensis was estimated to be 0.9-1.2 million years ago. PMID- 12589078 TI - Stages of institutional review board activities. PMID- 12589079 TI - Current status of the institutional review boards in Korea: constitution, operation, and policy for protection of human research participants. AB - The institutional review board is crucial to ensure the scientific and ethical quality of human participant research. This paper analyzes a survey on the current constitution and operation of institutional review boards (IRBs) in Korea, conducted by the Korean Association of Institutional Review Boards in April 2002. Out of 74 IRBs, 63 responded to the survey (85.1% response rate). IRB membership has a male-to-female ratio of approximately 80:20, a predominance of male clinicians (60%) and an underrepresentation of community people unaffiliated to the institutions (less than 10%). Most IRBs (around 80%) confine the scope of their reviews to the clinical evaluation of drugs or devices, leaving the remaining areas of research involving human participants untouched. As their role is limited, the majority of IRBs do not operate actively: 72% of responding IRBs reviewed less than one protocol per month in 2001. Sixty two percent of institutions have never discussed the need for insuring research participants' risks or making indemnity arrangements. This survey reveals many shortcomings and points for improvement by the institutional support bodies, including the need to establish regular education programs for IRB members and investigators. PMID- 12589080 TI - Recently occurring adult tetanus in Korea: emphasis on immunization and awareness of tetanus. AB - Since a nationwide childhood vaccination with tetanus toxoid, tetanus has become a rare disease in Korea. However, we recently experienced 17 cases of adult tetanus in a university hospital during a 21-month period. Seventy percent of the patients were female, and the mean age was 63 yr (range, 29-87). The majority (88.2%) of the patients did not get primary vaccinations for tetanus and decennial tetanus-diphtheria toxoid booster. Most patients (88.2%), who sustained acute injury, did not seek medical care for their wounds or did not receive the prophylaxis for tetanus. Tetanus was found most frequently among farmers. Tetanus was diagnosed initially only in 53% of patients. The case-fatality ratio was 23.5%. These cases show that recently occurring tetanus in Korea is a disease, affecting the elderly and the female who may have a lower immunity against tetanus, and the farmers who are likely to be exposed to Clostridium tetani. In addition, diagnosis of tetanus is often delayed in area where cases are seen infrequently. Therefore, improved education among patients and physicians, emphasis of anti-tetanus immunization and awareness of tetanus respectively, may be essential for the prevention of disease and the reduction of its mortality. PMID- 12589081 TI - Combined administration of glutamine and growth hormone synergistically reduces bacterial translocation in sepsis. AB - We investigated the combined effect of glutamine (GLN) and growth hormone (GH) on bacterial translocation (BT) in sepsis. After single intraperitoneal injection of lipopolysaccharide (10 mg/kg), 48 rats were divided randomly into four groups of 12 animals each: the control group received chow orally; the GLN group received chow plus 10% GLN; GH group received chow plus GH; and the GLN/GH group received chow, 10% GLN, and GH. Twenty-four and 96 hr later, rats were sacrificed. Portal blood culture, bacterial colony counts of cultured mesenteric lymph nodes, mucosal thickness, malondialdehyde (MDA), and glutathione (GSH) levels in the gut mucosa were measured. There was no significant change of the rate of portal blood culture between all treatment groups at 24 and 96 hr. At 24 hr, the rats receiving combined treatment of GLN and GH showed lower bacterial colony counts and mucosal MDA levels than the control rats, and higher mucosal GSH levels than the control and GLN-treated rats. At 96 hr, rats treated with both GLN and GH exhibited lower bacterial colony counts and mucosal MDA levels, and higher mucosal thickness and GSH levels than control, GLN, or GH-treated rats. This study suggests that the combination of GLN and GH may synergistically reduce BT over time in sepsis. PMID- 12589082 TI - Nicardipine augments local myocardial perfusion after coronary artery reperfusion in dogs. AB - Nicardipine is a potent coronary and systemic vasodilator without depression of ventricular function. We investigated the changes in local myocardial perfusion (LMP) according to the nicardipine administration after coronary reperfusion in a beating canine model. A Doppler probe was placed around the left anterior descending coronary artery (LAD) and thermal diffusion microprobe was implanted in the myocardium perfused by the exposed LAD. To define the nicardipine effects, we compared the two groups (control group, n=7 vs nicardipine group, n=7). In nicardipine group, 5 microgram/kg/min nicardipine was infused continuously. After the release of the LAD occlusion, LAD blood flow were increased compared to the baseline of both groups. However, there was no difference between groups in the LAD blood flow. The LMP after LAD reperfusion did not recover to the baseline level until 30 min after LAD reperfusion in control group (74%, 52% and 70% at 10, 20 and 30 min after LAD reperfusion, respectively). In nicardipine group, however, the LMP recovered to the baseline level at 20 min (99%), and increased more than the baseline level at 30 min (141%) after LAD reperfusion. Our findings suggest that the nicardipine augments the LMP following the release of a coronary occlusion. PMID- 12589083 TI - Clinical usefulness of the hematopoietic progenitor cell counts in predicting the optimal timing of peripheral blood stem cell harvest. AB - Although enumeration of CD34+ cells in the peripheral blood (PB) on the day of apheresis predicts the quantity of those cells collected, the flow cytometric techniques used are complex and expensive, and several hours are required to obtain the result in the clinical practice setting. The Sysmex SE-9000 automated haematology analyzer provides an estimate of immature cells, called hematopoietic progenitor cells (HPC). The aim of this study was to evaluate the clinical usefulness of HPC in predicting the optimal timing of peripheral blood progenitor cells (PBPC) harvest. Studies were performed on 628 aphereses from 160 patients with hematologic or solid malignancies. Spearman's rank statistics was used to assess correlation between HPC, WBC, mononuclear cells (MNC), and CD34+ cells. A receiver operating characteristic (ROC) curve was drawn for cutoff value of HPC, and predictive values of the chosen cutoff value of HPC for different target CD34+ cell collections were calculated. The PB HPC had a stronger correlation (rho=0.592, p<0.001) with collected CD34+ cells than did PB WBC and PB MNC. The ROC curve showed that the best cutoff value of HPC was 50 x 10(6)/L for the target CD34+ cells > or =1 x 10(6)/kg with sensitivity of 75%. Positive and negative predictive values of HPC > or =50 x 10(6)/L for CD34+ cells > or =1 x 10(6)/kg were 59.7% and 81.1%, respectively. In the clinical practice setting, applying variable cutoff values of HPC would be a useful tool to predict the optimal timing of PBPC collection. PMID- 12589084 TI - Chemotaxis of blood neutrophils from patients with primary ciliary dyskinesia. AB - Primary ciliary dyskinesia is characterized by chronic upper and lower respiratory infections which are caused by the grossly impaired ciliary transport. Since the cilia and neutrophils both utilize microtubular system for their movement, it has been speculated that neutrophil motility such as chemotaxis might be impaired in patients with primary ciliary dyskinesia. Neutrophils were purified from whole blood from 16 patients with primary ciliary dyskinesia and from 15 healthy controls. Chemotactic responses of neutrophils to leukotriene B(4) (LTB(4)), complement 5a (C5a), and formylmethionylleucylphenylalanine (fMLP) were examined using the under agarose method. The chemotactic differentials in response to LTB4, C5a, and fMLP in neutrophils from the patient group were significantly lower than the corresponding values in neutrophils from the control group (p<0.05 for all comparisons). The difference in chemotactic index between the two groups was statistically significant for LTB4 and fMLP (p<0.05 for both comparisons), but not for C5a (p=0.20). Neutrophils from patients with primary ciliary dyskinesia showed a decreased chemotactic response as compared with those from normal subjects. It is concluded that the increased frequency of respiratory tract infection in patients with primary ciliary dyskinesia is possibly due to the defective directional migration of neutrophils, as well as to the defective mucociliary clearance of the airways. PMID- 12589085 TI - Enhanced serum neutrophil chemotactic activity was noted in both early and late asthmatic responses during lysine-aspirin bronchoprovocation test in ASA sensitive asthmatic patients. AB - To investigate the pathogenic mechanism of late asthmatic response in comparison to early asthmatic response, changes of serum neutrophil chemotactic activity (NCA) using the Boyden chamber method and histamine level using the automated fluorometric analyzer were observed in 13 aspirin (ASA)-sensitive asthma subjects (group I: 7 early responders and group II: 6 dual responders) during lysine aspirin bronchoprovocation test (L-ASA BPT). Sera were collected before, and 30 min and 240 min after L-ASA BPT. Serum NCA increased significantly after 30 min (p=0.02) and decreased significantly at 240 min (p=0.02) in group I, while serum NCA of group II increased significantly at 30 min (p=0.04), tending to increase further up to 240 min with no statistical significance. NCA at 240 min in group II subjects was significantly higher than baseline NCA (p=0.02). The serum NCAs collected before and 240 min were significantly higher in group II than in group I (p<0.05, respectively). There were no significant changes in serum histamine levels during L-ASA BPT in both groups. NCA derived from mast cell may contribute to the development of early asthmatic response induced by L-ASA inhalation. There may be a possible involvement of NCA derived from mononuclear cells during late asthmatic response. PMID- 12589086 TI - The validity of the ISAAC written questionnaire and the ISAAC video questionnaire (AVQ 3.0) for predicting asthma associated with bronchial hyperreactivity in a group of 13-14 year old Korean schoolchildren. AB - To validate the prevalence rate of symptoms of asthma produced by the phase I ISAAC (International Study of Asthma and Allergies in Childhood) study, hypertonic saline challenge test was carried out during the phase II study at a year after the phase I study. For the phase II study, six middle schools from three cities in the phase I study were selected. Finally, 499 children who responded to both studies were analyzed. All subjects were asked to complete the written questionnaire (WQ) first, followed by a video questionnaire (AVQ 3.0) during the phase I study. Of the 499 children, only 19 (3.8%) were positive to the hypertonic saline bronchial challenge test. The degree of agreement between responses to the two corresponding questions "wheezing at rest" and "nocturnal wheeze" in the AVQ 3.0 and WQ were moderate and weak with a Kappa indices of 0.45 and 0.23, respectively. The question on "severe wheeze" in the AVQ 3.0 had the highest Youden's index among the five questions related to asthma symptoms in the previous 12 months, but its specificity was low whereas its sensitivity was 1.0. There was no consistency of priority between the two questionnaires in predicting bronchial hyperreactivity in a group of Korean schoolchildren. Therefore we need to develop more appropriate WQ or AVQ to compare the prevalences of asthma to other countries. PMID- 12589087 TI - No changing trends in incidence of gastric cardia cancer in Korea. AB - Although there is a controversy, several reports have shown that gastric cardia cancer has tended upwards in recent years. The aim of this study was to investigate the changing trend of stomach cancer according to the location of cancer and histologic type in Korea, where is known to be one of the most prevalent areas for gastric cancer. All the patients newly diagnosed as gastric cancer from 1991 through 2000 were recruited in Hallym University Medical Center. The periods were divided into the first (1991 through 1995) and the second (1996 through 2000) period. We analyzed the characteristics of cancer in the aspects of age, sex, tumor locations (cardia and non-cardia), and tumor grade of differentiation (well, moderate, and poor differentiated type). The number of cases was 2,395 and that of cardia cancer was 158 (6.6%). There was no evidence about the changes in the incidence of cardia cancer and histologic differentiation between two periods. The development of cardia cancer was not associated with the factors of age and sex with having no difference in the point of tumor grade. We could not find an upward tendency of cardia cancer development in recent 10 yr in Korea. PMID- 12589088 TI - Tissue plasminogen activator and plasminogen activator inhibitor type 1 gene polymorphism in patients with gastric ulcer complicated with bleeding. AB - Tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) may be involved in the pathogenesis of peptic ulcers through suppression of fibrinolysis. This study was designed to investigate associations of t-PA and PAI-1 genes with clinical features of the patients with bleeding gastric ulcers. Eighty-four patients with peptic ulcers and 100 controls were studied between January 1998 and April 2000. We used polymerase chain reaction and endonuclease digestion to genotype for 4G/5G polymorphism in the promoter region of the PAI-1 gene and the Alurepeat insertion/deletion (I/D) polymorphism in intron h of the t PA gene. Various clinical features, including lesion site, bleeding event, recurrence of ulcer, and rebleeding, were assessed using a multiple logistic regression model. The genotype distributions of both the t-PA and PAI-1 genes did not differ between the patient and control groups. The occurrence of the I/D or D/D genotype of t-PA was significantly higher in cases of duodenal ulcer (adjusted OR=4.39, 95% CI=1.12-17.21). When a dominant effect (i.e., 4G/4G or 4G/5G versus 5G/5G) of the 4G allele was assumed, the PAI-1 4G/4G genotype was independently associated with rebleeding after hemostasis (adjusted OR=5.07, 95% CI=1.03-24.87). Our data suggest that t-PA gene polymorphism is associated with duodenal ulcers, and that the PAI-1 gene may be a risk factor leading to recurrent bleeding after initial hemostasis. PMID- 12589089 TI - Heterozygous mutations of the gene for Kir 1.1 (ROMK) in antenatal Bartter syndrome presenting with transient hyperkalemia, evolving to a benign course. AB - Bartter-like syndrome encompasses a set of inherited renal tubular disorders associated with hypokalemic metabolic alkalosis, renal salt wasting, hyperreninemic hyperaldosteronism, and normal blood pressure. Antenatal Bartter syndrome, a subtype of Bartter-like syndrome, is characterized by polyhydramnios, premature delivery, life-threatening episodes of fever and dehydration during the early weeks of life, growth retardation, hypercalciuria, and early-onset nephrocalcinosis. Mutations in the bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2) and ATP-sensitive inwardly rectifying potassium channel (ROMK) of the thick ascending limb of Henle's loop have been identified in the antenatal Bartter syndrome. We report the identification of two heterozygous mutations of the gene for Kir 1.1 (ROMK) from an antenatal Bartter syndrome patient who presented at birth with mild salt wasting and a biochemical findings that mimicked primary pseudohypoaldosteronism type 1, such as hyperkalemia and hyponatremia, and evolved to a relatively benign course. We have identified amino acid exchanges Arg338Stop and Met357Thr in the gene exon 5 for ROMK by PCR and direct sequencing. Both mutations alter the C-terminus of the ROMK protein, and can affect channel function. PMID- 12589090 TI - Anti-heparin-platelet factor 4 antibody is a risk factor for vascular access obstruction in patients undergoing hemodialysis. AB - Since heparin is an anticoagulant commonly used in hemodialysis and the patients on hemodialysis are repeatedly exposed to heparin, heparin may be the cause of the development of heparin-dependent antibodies and thrombotic complications in patients on hemodialysis. The purpose of this study was to determine the prevalence and the clinical significance of the antibodies against heparin platelet factor 4 complexes as determined by enzyme immunoassay in patients on maintenance hemodialysis. The prevalence of anti-heparin-platelet factor 4 antibodies was higher in hemodialysis patients than in normal subjects (8.8 vs 0.0%, p<0.05). The number of past episodes of vascular access obstruction per year was significantly higher in the anti-heparin-platelet factor 4 antibody positive group than antibody negative group. Anti-heparin-platelet factor 4 antibody positive patients experienced more frequent vascular access obstructions than control subjects. In conclusion, anti-heparin-platelet factor 4 antibody might be a risk factor for vascular access obstructions in patients with end stage renal disease on maintenance hemodialysis. PMID- 12589091 TI - Parental concerns on the circumcision for elementary school boys: a questionnaire study. AB - To evaluate the parental concerns for elementary school boys (7-12 yr) on the circumcision, a randomly selected cross-sectional survey was performed in each elementary school from 16 urban wards in Busan. We asked 10,861 parents to answer the questionnaires on the circumcision such as the benefits and fallbacks of circumcision, proper time and knowledge of the surgery, and neonatal circumcision. The overall response rate to the questionnaire was 38.9%. The overall circumcision rate of elementary school boys was 43.2%, which increased from 18.7% at 7 yr old to 64.8% at 12 yr old. The significant reason for and against circumcision was "hygiene benefits (88.1%)" and "unreliable medical benefits (38.5%)", respectively. 74.9% of parents thought that elementary school age is the optimal time of circumcision. Only 11.2% of boys were circumcised during neonatal period. The main reason for parents to oppose neonatal circumcision was "their babies feel pain (35.8%)". About 50% of parents thought that circumcision will prevent medical diseases. Besides the medical basis, the circumcision is emerging as a kind of social custom in Busan. For parents making the decisions on the circumcision of their boys, physician or health care providers should provide helpful and honest facts about circumcision. PMID- 12589092 TI - Epidemiologic survey of head and neck cancers in Korea. AB - Head and neck cancers have never been systematically studied for clinical purposes yet in Korea. This epidemiological survey on head and neck cancer patients was undertaken from January to December 2001 in 79 otorhinolaryngology resident-training hospitals nationwide. The number of head and neck cancer patients was 1,063 cases in the year. The largest proportion of cases arose in the larynx, as many as 488 cases, which accounted for 45.9%. It was followed by, in order of frequency, oral cavity (16.5%), oropharynx (10.0%), and hypopharynx (9.5%). The male:female ratio was 5:1, and the mean age was 60.3 yr. Surgery was the predominant treatment modality in head and neck cancers: 204 (21.5%) cases were treated with only surgery, 198 (20.8%) cases were treated with surgery and radiotherapy, 207 cases (21.8%) were treated with combined therapy of surgery, radiotherapy, and chemotherapy. Larynx and hypopharynx cancers had a stronger relationship with smoking and alcohol drinking than other primary site cancers. Of them, 21 cases were found to be metastasized at the time of diagnosis into the lung, gastrointestinal tract, bone, or brain. Coexisting second primary malignancies were found in 23 cases. At the time of diagnosis, a total of 354 cases had cervical lymph node metastasis accounting for 42.0%. PMID- 12589093 TI - Survey and validation of the criteria for Behcet's disease recently used in Korea: a suggestion for modification of the International Study Group criteria. AB - The criteria employed for Korean Behcet's disease (BD) from January 1990 to December 2000 have been investigated, and the diagnostic validity for those criteria was determined. For the generation of a modified set of preliminary criteria from the International Study Group (ISG) criteria, the diagnostic values for individual feature of BD were calculated. The criteria by the Behcet's Disease Research Committee of Japan appeared to be widely employed with the ISG criteria. However, because the ISG criteria revealed a relatively valid outcome in Korea, the application of this criteria will be needed for the universal unification until the criteria with better performance comes out. On the other hand, the modified set of preliminary criteria that consisted of the clinical items with better results seemed to improve some pitfalls of the ISG criteria. Although that criteria showed better performance than the preexisting criteria, it should be necessary to validate its effectiveness in other areas. PMID- 12589094 TI - Comparison of transepidermal water loss, capacitance and pH values in the skin between intrinsic and extrinsic atopic dermatitis patients. AB - Atopic dermatitis (AD), with the prevalence rate of around 10 to 15%, is characterized by an intensely pruritic skin lesions with typical distribution and morphology. Recently, AD is divided into extrinsic type (ADe) and intrinsic type (ADi) according to the laboratory findings and associated diseases. ADe is well known for high IgE level, positive response to food- or aero-allergens, whereas ADi has clinically similar skin lesions and distribution patterns of AD with normal serum IgE levels, negative in vitro test for environmental or food allergens and without associated atopic diseases. To instrumentally evaluate the differences of skin involvement and functions between ADi and ADe, we checked the transepidermal water loss (TEWL), capacitance and pH in both types of childhood AD and age-matched control. The proportion of ADi was around 20% in all AD patients (10/51). Our experiment suggested possible differences between ADi and ADe. Antecubital fossa is a famous involvement site of childhood type of AD, where both types of AD patients showed higher TEWL and decreased capacitance. ADe patients showed increased TEWL in all sites and lower hydration in 4 sites, whereas ADi patients showed no significant differences of TEWL and hydration in forehead, cheek, and back of leg. PMID- 12589096 TI - Clinical usefulness of the two-site Semmes-Weinstein monofilament test for detecting diabetic peripheral neuropathy. AB - The present study was done to validate the two-site Semmes-Weinstein (SW) monofilament test in identifying patients at risk of lower-extremity complications in clinical setting. The SW monofilament test and nerve conduction study were conducted on type 2 diabetic patients (n=37) at Pusan National University Hospital in Korea. As the duration of diabetes mellitus was longer, neuropathy identified by nerve conduction study and complications of diabetes were more severe (p<0.01). The number of sites unable to perceive SW monofilament (p<0.001) was larger in patients with lower-extremity neuropathy symptoms than those without symptoms. Sensitivity and specificity at two sites (the third and fifth metatarsal head sites) were 93% and 100%, respectively. In conclusion, the two-site SW monofilament test was a sensitive, specific, simple, and inexpensive screening tool for identifying diabetic peripheral neuropathy in clinical setting. PMID- 12589095 TI - Eosinophil inflammation of nasal polyp tissue: relationships with matrix metalloproteinases, tissue inhibitor of metalloproteinase-1, and transforming growth factor-beta1. AB - Eosinophil and mast cell infiltrations are consistent findings in nasal polyp tissue. Previous studies have shown that matrix metalloproteinases (MMPs) may be involved in eosinophil infiltration in airway mucosa of asthmatic patients, and that transforming growth factor-beta1 (TGF-beta1) induces extracellular matrix deposition in nasal polyp tissue. The aim of this study was to evaluate the role of MMPs and tissue-inhibitor of metalloproteinase-1 (TIMP-1) in association with TGF-beta1, eosinophils and mast cell activation in nasal polyp tissue. Nasal polyp tissues from 20 patients who underwent polypectomies were collected and prepared into tissue homogenate. Eosinophil cationic protein (ECP) and tryptase levels were measured by CAP system (Pharmacia, Sweden). MMP-2, MMP-9, TIMP-1 and TGF-beta1 levels were measured by enzyme-liked immunosorbent assay. MMP-2 was the predominant form of MMPs, followed by MMP-9 and TIMP-1. There were significant correlations between ECP, and MMP-9, MMP-2, TGF-beta1 and tryptase, but not with TIMP-1. Significant correlations were noted between tryptase, and MMP-2, MMP-9, and TGF-beta1, but not with TIMP-1. Close correlations were noted between TGF beta1, and MMP-9 and MMP-2, but not with TIMP-1. MMP-2, MMP-9, and TGF-beta1 may contribute to eosinophil and mast cell migrations into nasal polyp tissue. PMID- 12589097 TI - Distribution of adenoviral vector in brain after intravenous administration. AB - The delivery of transgenes to the central nervous system (CNS) can be a valuable tool to treat CNS diseases. Various systems for the delivery to the CNS have been developed; vascular delivery of viral vectors being most recent. Here, we investigated gene transfer to the CNS by intravenous injection of recombinant adenoviral vectors, containing green fluorescence protein (GFP) as a reporter gene. Expression of GFP was first observed 6 days after the gene transfer, peaked at 14 days, and almost diminished after 28 days. The observed expression of GFP in the CNS was highly localized to hippocampal CA regions of cerebral neocortex, inferior colliculus of midbrain, and granular cell and Purkinje cell layers of cerebellum. It is concluded that intravenous delivery of adenoviral vectors can be used for gene delivery to the CNS, and hence the technique could be beneficial to gene therapy. PMID- 12589098 TI - Partial trisomy 2q(2q37.3-->qter) and monosomy 7q(7q34--->qter) due to paternal reciprocal translocation 2;7: a case report. AB - We report an unbalanced translocation involving chromosome 2 and 7 due to a balanced reciprocal translocation 2;7 in the father. The female fetus had a partial trisomy of the long arm of chromosome 2 with a partial monosomy of distal 7q. Ultrasound at the first trimester had indicated normal fetal anatomy, including normal intracranial structures. Parental karyotypes showed a paternal balanced translocation: 46,XY,t(2;7)(q37.3;-->q34). The unbalanced translocation in the fetus resulted in trisomy for 2q37.3 qter and monosomy for 7q34-->qter. Postnatal examination showed that the female abortus had a cleft lip and palate, and mild dysmorphic features. The clinical phenotype was in agreement with previous descriptions and allowed us to propose a fetal phenotype for this chromosomal abnormality. PMID- 12589099 TI - Electrical injury as a possible cause of sick sinus syndrome. AB - Electrical injury is a serious public health problem. Heart is one of the most frequently affected organs. Electrical injury can cause life-threatening cardiac complications such as asystole, ventricular fibrillation, and myocardial rupture. In this case report, we present a 20-yr-old male patient with sick sinus syndrome that developed years after electrical injury. PMID- 12589100 TI - Cervical epidural abscess secondary to aorto-duodenal fistula: a case report. AB - Although cervical epidural abscess is rare, it should be strongly suspected in any patient with unexplainable neck pain and fever, especially when the patient has a predisposing factor for this infectious process. The authors report a case of cervical epidural abscess in a 39-yr-old man with an aorto-duodenal fistula, which complicated the interposition of artificial graft for abdominal aortic aneurysm rupture, which had undertaken 40 months before. Timely detection and intervention rendered him a full neurological recovery. This extremely rare case is presented with a literature review. PMID- 12589101 TI - A case of primary esophageal B-cell lymphoma of MALT type, presenting as a submucosal tumor. AB - The primary esophageal lymphoma is extremely rare, and shows various morphologic characteristics. Only a single case of mucosa-associated lymphoid tissue (MALT) type lymphoma confined to the esophagus has been reported in the literature. A 61 yr-old man was referred to our hospital for evaluation of an esophageal submucosal tumor (SMT) that had been detected incidentally by endoscopy. He had a history of pulmonary tuberculosis with long-term anti-tuberculosis medication 15 yr before, and also had a history of syphilis, which had been treated one year before. He had been taking a synthetic thyroid hormones for the past 10 months because of an autoimmune thyroiditis. Endoscopy showed a longitudinal round and tubular shaped smooth elevated lesion, which was covered with intact mucosa and located at the mid to distal esophagus, 31 cm to 39 cm from the incisor teeth. Endoscopic ultrasonography (EUS) showed a huge longitudinal growing intermediate- to hypo-echoic mass located in the submucosal layer with internal small, various sized honeycomb-like anechoic lesions suggesting germinal centers. Subsequently, he underwent a surgery, which confirmed the mass as a primary esophageal low grade B-cell lymphoma of MALT type. PMID- 12589102 TI - Infravesical obstruction due to benign intraurethral prostatic cyst. AB - We report a case of symptomatic intraurethral prostatic cyst in a 42-yr-old man without clinical evidence of benign prostatic hyperplasia. The intraurethral cyst makes it unique from the all previously reported cases of prostatic cysts located medially within the prostate. Transurethral resection of the cyst with limited resection of the prostatic tissue at the base of the cyst was performed with successful resolution of voiding symptoms. PMID- 12589103 TI - Extraskeletal osteochondroma of the buttock. AB - Osteochondromas are common and typically arise from the metaphyseal ends of long bones. An osteochondral neoplasm of the soft tissue, which is a lesion of uncertain pathogenesis, is uncommon and usually arises from the synovial tissue in joints and tendon sheaths. Rarely, extraskeletal osteochondromas also arise outside of synovial compartments. Most of the reported cases were presented in the hands and feet, especially in the fingers. Here we describe a 44-yr-old female patient who presented with a pain in the left buttock. A well-defined osseous mass was detected in the buttock. It consisted of sharply demarcated, mature hyaline cartilage that was covered with a fibrous capsule, which changed gradually into cancellous bone, more pronouncedly at the center. The diagnosis of an extraskeletal osteochondroma should be considered when a discrete, ossified mass is localized in the soft tissues. A case of pathologically proven extraskeletal osteochondroma of the buttock is presented with a literature review, magnetic resonance imaging, and radiological findings. PMID- 12589104 TI - Successful combination therapy of cyclosporine and methotrexate for refractory polymyositis with anti-Jo-1 antibody: a case report. AB - Although corticosteroids have been the initial agent for the treatment of inflammatory myopathies (IM), immunosuppressive agents such as azathioprine, methotrexate, cyclophosphamide, or cyclosporine are commonly required to control the disease except mild cases. On the other hand, the efficacy of combination therapy of cyclosporine and methotrexate in severe rheumatoid arthritis has been proven without serious side effects. However, in treatment-resistant myositis, the experience of such a therapy is very limited, and has not been described in refractory polymyositis with anti-Jo-1 antibody. Here, we report a young female patient with recalcitrant polymyositis and anti-Jo-1 antibody who was successfully treated with the combination therapy of cyclosporine and methotrexate. At first, the myositis did not respond to several agents, such as corticosteroid, monthly pulse cyclophosphamide, azathioprine, or cyclosporine. Methotrexate was initially avoided as treatment regimen because of its potential pulmonary toxicity in the case with preexisting lung disease. PMID- 12589105 TI - Tubular aggregate myopathy: a case report. AB - We report a first Korean case of presumably dominantly inherited primary tubular aggregate myopathy in a 19-yr-old man, who presented with slowly progressive proximal muscle stiffness and weakness. In hematoxylin and eosin stain, it showed subsarcolemmal, or central pale basophilic granular vacuoles, which stained red with modified Gomori's trichrome and intensive blue with nicotinamide adenonine dinucleotide-tetrazolium reductase, respectively. Ultrastructurally, aggregates of 60 nm-sized hexagonal tubules were found in both type 1 and type 2 fibers. We briefly review the pathologic findings of the previously reported cases of tubular aggregate myopathy and discuss the possible pathogenesis of this disease. We briefly discuss the possible pathogenesis of sarcoplasmic reticulum and review the ultrastructural characteristics. PMID- 12589107 TI - Conn's syndrome associated with Behcet's disease. AB - A 39-yr-old woman, who had been treated for Behcet's disease for 4 yr, was admitted for further investigation of recently identified hypokalemia and hypertension. Suppressed plasma renin activity with elevated plasma aldosterone concentration and an anomalous postural decrease in plasma aldosterone were observed. An abdominal CT scan revealed a right adrenal mass. The patient was diagnosed with Conn's syndrome. The association of Conn's syndrome with Behcet's disease was thought to be coincidental. To our knowledge, this is the first case of Conn's syndrome associated with Behcet's disease. PMID- 12589106 TI - A novel mutation (C67Y)in the NOTCH3 gene in a Korean CADASIL patient. AB - We report a 52-yr-old Korean woman with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) whose diagnosis was confirmed by skin biopsy and the presence of a novel mutation in the NOTCH3 gene. The patient's clinical features were rather unusual in that 1) clinical presentations were only two episodes of stroke and mild dementia unaccompanied by mood disturbances or migraine, and 2) there was no family history. Brain MRI showed T2 hyperintensities in both temporal pole areas in line with the recent suggestion by O'Sullivan et al. that the abnormality could be a radiologic marker of CADASIL. FDG-PET also showed a hypometabolism in the temporal pole areas with an abnormal finding on MRI in addition to the hypometabolism in cortical and subcortical regions. We could learn from this case that CADASIL may be included in the differential diagnoses in patients with vascular dementia associated with a small vessel disease, even in the absence of a family history, especially when there are no known stroke risk factors and when the MRI shows T2 hyperintensity in the temporal pole regions. PMID- 12589108 TI - Impact of the human genome project on pediatric endocrinology. AB - A genomic approach to human biology involves examination of the entire complement of human genes and their protein products in contradistinction to the conventional phenomena-driven examination of individual components, one at a time. This approach may have limitations, but it also has the potential to increase our knowledge by an order of magnitude or more. It is expected that it will leave no field of biology and medicine unaffected, and this includes endocrinology. The structural and functional aspects of the human genome are reviewed, and the nature of the novel knowledge from the genome effort (acquired or expected) is described. More importantly, an overview is given of new ways of thinking and new approaches to endocrine research using genomic concepts and tools. Although these research breakthroughs have relatively few applications in clinical practice at the present moment, the clinician must expect drastic changes in diagnostics and therapeutics in the next decade or two, and endocrine- specific examples of such applications are given. Finally, the question is raised of social and ethical issues that these developments are generating. PMID- 12589109 TI - Emergence and evolution of the circadian rhythm of melatonin in children. AB - OBJECTIVE: To assess the age at which the circadian rhythm of melatonin begins. METHODS: 55 children, divided into groups from the neonatal period to 24 months of life, were studied. Urine samples were taken from 28 newborn babies to measure 6-sulfatoxymelatonin (aMT6s). Salivary samples were collected from infants (27 cases), to measure melatonin (aMT). aMT was measured by RIA and aMT6s by ELISA using commercial kits. Changes in the levels of aMT6s and aMT were evaluated using the Friedman test and Wilcoxon matched pair test. RESULTS: The group aged 27-41 days showed statistically significant differences in daily aMT6s and aMT concentrations. The highest values were always found between 24.00 and 8.00 h. This day/night difference persisted from 2-3 to 13-24 months of age. CONCLUSION: The data indicate that the circadian melatonin rhythm appears at the end of the neonatal period and persists thereafter. PMID- 12589110 TI - Disproportionately elevated fasting proinsulin levels in normoglycemic patients with thalassemia major are correlated to the degree of iron overload. AB - OBJECTIVE: To analyze the secretion of the insulin precursor proinsulin in patients with beta-thalassemia and its possible relation to iron overload. METHODS: We assessed fasting proinsulin, insulin, C-peptide and glucose levels from 34 patients with beta-thalassemia and 33 healthy controls. The correlation to age, body mass index, hepatic iron concentration, serum ferritin and serum AST was analyzed. RESULTS: Fasting proinsulin (p < 0.002) and proinsulin-to-insulin ratio (p < 0.02) were significantly increased in patients with thalassemia irrespective of the degree of glucose tolerance. They correlated positively to serum ferritin, liver iron, patient age and serum AST (all p < 0.05). CONCLUSIONS: Disproportionately elevated proinsulin levels in thalassemic patients indicate early beta-cell dysfunction due to siderosis. An additional biological significance of hyperproinsulinemia and its possible ability to predict long-term iron toxicity in these patients remain to be clarified. PMID- 12589111 TI - One-year spinal bone change in pre- and perimenopausal Japanese women. A prospective observational study. AB - BACKGROUND: This prospective observational study was designed to determine whether the bone mineral density (BMD) of the lumbar spine decreases before menopause. METHODS: The change in BMD of the second through fourth lumbar vertebrae (delta%L2-4BMD) over the course of 12 months was measured in 197 pre- and perimenopausal Japanese women aged 48.2 +/- 2.3 (mean +/- SD) years. RESULTS: Overall, delta%L2-4BMD decreased significantly, with a greater decrease seen in perimenopausal women. This group also had a significantly higher level of FSH (p < 0.05, t = 7.356), a significantly lower level of estradiol (p < 0.05, t = 4.245), and significantly higher levels of the bone metabolic markers, alkaline phosphatase (p < 0.05, t = 3.841), calcium (p < 0.05, t = 3.939), and osteocalcin (p < 0.05, t = -3.295). Overall, there was a significant positive correlation between osteocalcin and delta%L2-4BMD (r = -0.194, p = 0.0479). CONCLUSION: A subset of perimenopausal women with transient decreases in ovarian function that do not respond to increased FSH may be at increased risk for abnormally low BMD, and may benefit from early management of bone mass. PMID- 12589112 TI - Suppression of TSH in congenital hypothyroidism is significantly related to serum levels and dosage of thyroxine. AB - AIM: To assess thyrotropin (thyroid-stimulating hormone; TSH) suppression and serum thyroxine (T(4)) concentrations in infants with congenital hypothyroidism in relation to T(4) dose and pretreatment parameters. METHOD: A retrospective study of all cases treated in a single centre since neonatal screening began was performed. RESULTS: In 54 infants treated with a mean daily T(4) dose of 9.8 microg/kg, the TSH concentration was suppressed (<6 mU/l) in 65% of the cases by 6 months with the serum T(4) level at the upper end of the infant reference range. Infants who suppressed their TSH later did not differ in pretreatment serum TSH or T(4) concentration. T(4) dose and serum T(4) level were lower in infants whose TSH was not suppressed. CONCLUSIONS: TSH suppression in congenital hypothyroidism is significantly related to serum levels and dosage of T(4). We suggest that a delay in TSH suppression is mainly due to undertreatment. PMID- 12589113 TI - Effect of heavy increase in training stress on the plasma leptin concentration in highly trained male rowers. AB - AIM: To characterize the effects of a heavy increase in training stress followed by a reduced stress on fasting plasma leptin levels in highly trained male rowers. METHODS: 12 rowers underwent a 3-week period of maximally increased training stress followed by a 2-week tapering period. RESULTS: A mean 22% increase in training stress caused a significant decrease (by 8%) in the leptin concentrations. A further increase in training stress by 25% significantly reduced leptin further by 35%. A 1st tapering week, during which the training stress was rapidly reduced by approximately 50%, significantly increased the plasma leptin concentrations by 29%. Plasma leptin was significantly increased further (by 4%) during the 2nd tapering week. CONCLUSION: Leptin is sensitive to pronounced changes in training stress in highly trained male rowers. PMID- 12589114 TI - The serum concentration of estradiol after embryo transfer and the decline from preovulatory levels may influence the success of IVF treatment. AB - AIM: To establish the influence that serum estradiol concentrations prior to oocyte retrieval and 3 days after embryo transfer have on the establishment of in vitro fertilization (IVF) pregnancy. METHOD: Preovulatory (day-0) and luteal phase (day-6) estradiol concentrations were retrospectively analyzed in 310 infertile patients, undertaking 1st-cycle conventional IVF or intracytoplasmic sperm injection. RESULTS: The IVF treatment success is significantly reduced in patients with an estradiol level <600 pg/ml and also when a rapid decline in luteal-phase estradiol concentrations from preovulatory concentrations (day 0:day 6 ratio) was exhibited. CONCLUSION: A day 0:day 6 estradiol ratio >5 and a serum estradiol concentration <600 pg/ml may adversely impact on the establishment of pregnancy in IVF. PMID- 12589115 TI - Dynamic change in plasma leptin level during the perioperative period. AB - OBJECTIVE: To examine the association of plasma leptin level with insulin, which is known as a metabolic regulator of leptin, and various stress-related factors during the perioperative period. METHODS: Thirty-one patients undergoing gastrectomy were enrolled and blood samples were obtained preoperatively, intraoperatively, immediately after operation, and on the first and second postoperative days. RESULTS: The plasma leptin level showed a triphasic response. Immediately after operation, the leptin level was the lowest, while the serum cortisol and interleukin-6 (IL-6) levels had their peak (PHASE 1). On the first postoperative day, the leptin level had its peak with the serum cortisol and IL-6 level remaining elevated (PHASE 2). On the second postoperative day, while the serum cortisol and IL-6 levels still remained elevated, the plasma leptin level fell to the preoperative value (PHASE 3). The plasma leptin level correlated well with the insulin level at all time points during the study period. CONCLUSION: Our results suggest a possible role of leptin in harmonizing neuroendocrine and immune responses with energy balance. PMID- 12589116 TI - Hoffman's syndrome: muscle stiffness, pseudohypertrophy and hypothyroidism. AB - Primary hypothyroidism is a chronic and insidious disease caused by failure of thyroid hormone production. We observed a 38-year-old woman admitted to our hospital due to progressive proximal weakness, muscle pain and fatigue during mild exercise. Laboratory tests showed features of rhabdomyolysis and hypothyroidism. After examination of the thyroid, we reached a diagnosis of Hashimoto's thyroiditis and hypothyroid myopathy. Hypothyroidism should be considered as a differential diagnosis of creatine kinase elevation; actually, neuromuscular symptoms and signs occur in most newly diagnosed patients with thyroid diseases. Hypothyroidism presenting as muscle stiffness and pseudohypertrophy is called 'Hoffman's syndrome'. PMID- 12589117 TI - The heart in human dystrophinopathies. AB - Dystrophinopathies are due to mutations in the dystrophin gene on chromosome Xp21.1 and comprise the allelic entities Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) and X-linked dilative cardiomyopathy (XLDCM). In all three entities, the heart is affected to various degrees, depending on the stage of the disease and the type of the mutation (cardiac involvement, CI). The pathoanatomic evidence of CI in dystrophinopathies is the replacement of myocardium by connective tissue or fat. In DMD/BMD, the left ventricular posterobasal and lateral walls are most extensively affected, sparing the right ventricle and the atrium. Degree and dynamics of CI vary among the three entities. In DMD/BMD, CI usually remains subclinical in the early stages of the disease. Typical initial manifestations of CI in DMD/BMD are sinus tachycardia, tall R1 in V1, prominent Q in I, aVL, V6 or in II, III, and aVF, increased QT dispersion and possibly autonomic dysfunction. Initially, echocardiography is normal or shows regional wall motion abnormalities in areas of fibrosis. With spreading of fibrosis, left ventricular dysfunction and ventricular arrhythmias additionally occur. In the final stages of the disease, systolic function may lead to heart failure and sudden death. Subclinical or clinical CI is present in about 90% of the DMD/BMD patients but is the cause of death in only 20% of the DMD and 50% of the BMD patients. XLDCM is a rapidly progressive, almost exclusively myocardial disorder, starting in teenage males as heart failure due to dilative cardiomyopathy (CMP), leading to death from intractable heart failure within 1-2 years after diagnosis. Therapy of arrhythmias and CMP in all three disorders follows the established cardiological recommendations. Due to its protective effect, ACE inhibitors are recommended already at the early stages of the disease. Beta-blockers may be an additional option if indicated. PMID- 12589118 TI - Cost-effectiveness of spinal cord stimulation versus coronary artery bypass grafting in patients with severe angina pectoris--long-term results from the ESBY study. AB - The present study is a 2-year follow-up of the 104 patients participating in the ESBY study (Electrical Stimulation versus Coronary Artery Bypass Surgery in Severe Angina Pectoris), a randomised prospective study including patients with increased surgical risk and no prognostic benefit from revascularisation. Hospital care costs, morbidity and causes of death after spinal cord stimulation (SCS) and coronary artery bypass grafting (CABG) were assessed, as well as the complication rate of SCS treatment. SCS proved to be a less expensive symptomatic treatment modality of angina pectoris than CABG (p < 0.01). The SCS group had fewer hospitalisation days related to the primary intervention (p < 0.0001) and fewer hospitalisation days due to cardiac events (p < 0.05). The groups did not differ with regard to causes of death. There were no serious complications related to the SCS treatment. PMID- 12589119 TI - Mechanical complications after myocardial infarction reliably predicted using C reactive protein levels and lymphocytopenia. AB - We assessed the accuracy of C-reactive protein (CRP) levels and lymphocyte counts to predict a mechanical complication (MC) after myocardial infarction (MI). Within 10 years, we identified 36 patients with 39 echocardiographically confirmed MC within 30 days of MI: ventricular septal defect (17 cases), papillary muscle rupture (10 cases), and left ventricular free wall rupture (12 cases). They were compared to 41 controls with an uncomplicated hospital course after MI. Peak CRP levels and minimum relative lymphocyte counts obtained within 96 h of the acute MI (AMI) and before diagnosis of the complication were compared with clinical parameters. Prior to the MC, peak CRP levels were significantly higher (p < 0.001) and relative lymphocyte counts lower (p < 0.001) than in controls while creatine kinase levels did not differ (p = nonsignificant). Using multivariate logistic regression, the following score was identified to have excellent prognostic significance for MC: CRP (mg/l) - 10 x Lyc (%). The area under the receiver-operating characteristic curve was 0.90 +/- 0.05 (p < 0.001). Combined use of CRP levels and relative lymphocyte counts may be helpful in accurately predicting an MC after AMI and should therefore be routinely assessed. PMID- 12589120 TI - Myocardial angiogenesis resulting in functional communications with the left cavity induced by intramyocardial high-frequency ablation: histomorphology of immediate and long-term effects in pigs. AB - A previous study in rabbit hearts demonstrated the feasibility of creating transmyocardial channels by application of temperature-regulated high-frequency (HF) energy. Feasibility of creating non-transmural channels using a transvascular approach was tested in 14 pigs which were followed for 1 h, and 3 and 9 weeks (group A, B and C). Myocardial channels were found to be highly reproducible and patent in more than two thirds in group A. Channels were replaced by connective tissue during follow-up which contained newly formed small vessels. A functional communication between the left ventricular cavum and newly formed vessels of the channel remnants could be demonstrated in one heart of group C. Thus, intramyocardial HF ablation can be performed with high reproducibility to induce local angiogenesis. PMID- 12589121 TI - Age and sex differences in the treatment of patients with initial acute myocardial infarction: a community-wide perspective. AB - PURPOSE: The goal of this observational study was to examine overall and age specific differences between women and men in the use of five beneficial cardiac medications in patients hospitalized with acute myocardial infarction (AMI) from a community-wide perspective. The objectives of our study were to determine whether women are treated differently than men and whether age acts as a potential effect modifier of any observed associations. SUBJECTS: A total of 2,461 women and 3,454 men with validated AMI comprised the study sample. METHODS: Our study was an observational investigation of metropolitan Worcester (Mass., USA) residents who were hospitalized with initial AMI in all area hospitals during 12 1-year periods between 1975 and 1999. Four age-specific subgroups (<55, 55-64, 65-74 and > or =75 years) were studied. RESULTS: Differences in the use of angiotensin-converting enzyme (ACE) inhibitors, aspirin, beta-blockers, lipid lowering medications and thrombolytic agents during hospitalization for AMI were examined. The results of a multivariable regression analysis indicated that women were significantly less likely to receive aspirin and ACE inhibitors as compared to men. There were no significant gender differences in the prescribing of the other cardiac medications. Increasing age in both women and men was associated with a reduced likelihood of receiving effective cardiac therapies including aspirin, beta-blockers, lipid-lowering therapy and thrombolytic agents. CONCLUSIONS: These data suggest that the reasons for the marked age-related differences, in men and women, in the use of cardiac medications be more systematically explored. Previously observed gender differences in the management of patients with AMI essentially no longer exist. PMID- 12589123 TI - Mechanisms of syncope and Stokes-Adams attacks in bradyarrhythmias: asystole and torsade de pointes. PMID- 12589122 TI - Early use of beta-blockers is associated with attenuation of serum C-reactive protein elevation and favorable short-term prognosis after acute myocardial infarction. AB - BACKGROUND: We have reported that a marked elevation in serum C-reactive protein (CRP) level is a predictor for infarct expansion and cardiac rupture after AMI. Although beta-blockers prevent cardiac rupture after AMI, their effect on serum CRP elevation has not been determined. METHODS: We studied a total of 154 patients with first Q-wave AMI. Patients complicated by pump failure were excluded from this study. Eighty-two patients received beta-blocker treatment within 24 h of the onset of AMI, while 72 patients received no beta-blocker treatment. Peak serum creatine kinase (CK) and CRP levels were determined by serial measurements. RESULTS: There was no difference between the groups according to age, sex, coronary risk factors, pre-infarction angina, infarct site, prior use of cardiovascular drugs, use of revascularization therapy, and prevalence of multivessel disease. Beta-blocker treatment was associated with a lower peak CRP level (6.9 +/- 6.1 vs.10.8 +/- 9.3 mg/dl, p = 0.002), a shorter duration from the onset to the peak CRP level (2 +/- 1 vs. 3 +/- 2 days, p < 0.0001), a lower incidence of cardiac rupture (p = 0.03) and lower in-hospital cardiac mortality (p = 0.02), despite similar peak CK levels. CONCLUSION: The early use of beta-blockers is associated with decreased serum CRP level and a favorable clinical outcome after first Q-wave AMI, suggesting some beneficial effects of beta-blockers on infarct healing after AMI. PMID- 12589124 TI - Left atrial myxoma presenting as acute myocardial infarction in a child. PMID- 12589125 TI - Cortical blindness: a rare but dramatic complication following coronary angiography. PMID- 12589126 TI - Wolff-Parkinson-White syndrome: pseudo myocardial infarction and pseudo ventricular hypertrophy patterns on electrocardiogram. PMID- 12589129 TI - Ethics and rationale of living-donor liver transplantation in Asia. AB - Living-donor liver transplantation took root in Asia as a natural result of circumstances, because the supply of organs from the cadaveric pool remained scarce over the years. In contrast to Western countries, the majority of organs for liver transplantation in Asia come from live donations. As the expertise of the transplant teams grows, patient outcomes improve, and public awareness increases, the option of live donation of the liver is increasingly chosen. Although no live liver donor death has yet been reported from Asia, the risk is not eliminated and remains a major consideration in the potential donor's decision to donate. The low morbidity and mortality rate could be attributed to the extensive experience of surgeons in liver surgery, because surgical liver disease is highly prevalent in Asia. Although the donor risk is estimated to be low, live organ donation should be absolutely voluntary, with consent given on the basis of unbiased information and chosen only when the option for obtaining a cadaveric graft is practically nil. It is only under these conditions that living donor liver transplantation should be perpetuated. Although the disease-donation transplantation process involves a complex interplay of psychosocial and family dynamics, the potential candidate's perception will necessarily depend on the surgeon's explanation. The ethical soundness of the practice of living-donor liver transplantation rests primarily on the ones who deliver the service. PMID- 12589130 TI - Living-donor liver transplantation: 12 years of experience in Asia. AB - Living-donor liver transplantation has provided a solution to the severe lack of cadaveric grafts for the replacement of livers afflicted with end-stage cirrhosis, fulminant disease, or inborn errors of metabolism. The pioneering experience in Japan in the early 1990s helped open wide the avenues of a new branch of science that is technically demanding and whose benefits are clearly dramatic. The need for alternative sources of liver grafts was common to the entire Asian region and, fortunately, the option of obtaining partial liver grafts from live donors had already become tenable. By the second half of the past decade, living-donor liver transplant programs had been successfully established in Hong Kong, Taiwan, and Korea. More than 1,500 cases have been performed over a 12-year period. This report describes the cumulative experience in living-donor liver transplantation in Asia on the basis of data from five major liver transplant centers. PMID- 12589131 TI - Complications and long-term outcome of living liver donors: a survey of 1,508 cases in five Asian centers. AB - A multicenter survey was conducted regarding the safety and outcome of living liver donors in Asia. Five Asian liver transplant centers reported a total of 1,508 living donor liver transplantations (LDLT) performed between January 1990 and December 2001. The recipients consisted of 766 adults and 742 children. The graft types were left lateral or extended left lateral segment in 605, left lobe in 334, right lobe or right lateral sector in 561, and not classified in eight cases. The donor blood loss was less than 1,000 mL in 94.1% of the cases, and 0.53% of the donors received banked blood transfusion. The overall donor complication rate was 15.8%, and 1.1% of donors underwent reoperation. The complication rate was higher in right lobe (28%) than in left lateral segment (9.3%) or left lobe (7.5%) donors. In particular, right lobe donors had more serious complications such as cholestasis (7.3%), bile leakage (6.1%), biliary stricture (1.1%), portal vein thrombosis (0.5%), intra-abdominal bleeding (0.5%), and pulmonary embolism (0.5%). There was no hospital mortality, but there was one late donor death 3 years after operation. Long-term follow-up beyond 3 months was available in 15.1% of the donors only. The data from the multicenter study set the standard for the safety of living liver donors in high-volume liver transplant centers in Asia and provide further justification for the continued application of LDLT in the face of the critical organ shortage in Asia. The long term outcome of liver donation, however, remains unknown, and transplant centers should continue their follow-up of donors. PMID- 12589132 TI - Evaluation of living liver donors. AB - Preoperative evaluation of donors for living-donor liver transplantation aims to select a suitable donor with optimal graft quality and to ensure donor safety. There are minor variations in the donor selection process among different centers, but the safety of the donor remains central to the entire process. The potential donors are evaluated in a stepwise manner including medical, physical, laboratory, psychosocial, and imaging assessment to disqualify unsuitable donors as early as possible in the evaluation process. The main goal of the imaging study is to provide an accurate picture of liver vascular anatomy and liver volume measurement for surgical guidance or for exclusion of unsuitable donors. All imaging studies can now be obtained using noninvasive modalities, thereby decreasing the risk associated with the donor evaluation process. This article describes the donor selection practice in our center including the details of the imaging evaluation. PMID- 12589133 TI - Small-for-size graft problems in adult-to-adult living-donor liver transplantation. AB - The number of adult patients undergoing living donor liver transplantation (LDLT) has recently increased. According to a recent Japanese survey, the 5-year survival rate in adults after surgery is only 69.7%, which is significantly poorer than that in pediatric series. Small-for-size grafts remain a problem in adult LDLT. The most commonly used liver graft for adult patients has shifted from the left liver to the right, which alleviates the problem of size disproportion. Right hepatectomy, however, increases the extent of the donor operation and raises an important ethical issue in LDLT. Patients who truly need a right liver graft should be carefully selected using evidence-based criteria. PMID- 12589134 TI - Living-donor liver transplantation using the left liver, with special reference to vein reconstruction. AB - The authors describe their techniques for hepatic vein reconstruction, devised for safe living-donor liver transplantation using a left liver graft. End-to-end anastomosis of the hepatic veins is performed to prevent an outflow occlusion. To ensure adequate hepatic venous flow, it is necessary to obtain a wide ostium and sufficient length of the hepatic vein for anastomosis, which should be secured by venoplasty of the hepatic veins of the graft and the recipient. A left liver with a caudate lobe graft is useful for overcoming the problem of a small graft. Reconstruction of the short hepatic vein of the caudate lobe may allow this portion to regenerate at the same rate as the left liver. In a left liver graft without the trunk of the middle hepatic vein, reconstruction of a tributary of this vein might be necessary to prevent graft congestion in segment IV. Color Doppler ultrasonography or a hepatic arterial clamping test should be performed in donor hepatectomy to evaluate the need for middle hepatic vein reconstruction. PMID- 12589135 TI - Right anterior sector drainage in right-lobe live-donor liver transplantation. AB - The necessity of providing venous drainage for the right anterior sector of a right-lobe graft in adult-to-adult right-lobe live-donor liver transplantation has been controversial. Inclusion of the middle hepatic vein in the right-lobe graft to ensure better early graft function is also under debate. This report summarizes the views of five Asian centers on the necessity of providing venous drainage to the right anterior sector in a right-lobe graft as presented at the Asian Living Donor Transplantation Symposium 2002. All five centers recognize the importance of adequate drainage of the right anterior sector, but they adopt different approaches in including the middle hepatic vein in the graft. Tokyo University uses an occlusion test of the right hepatic artery and middle hepatic vein to define whether the right anterior sector is dusky or regurgitation of blood flow is present in the right anterior portal vein before the decision is made for middle hepatic vein reconstruction. The Asan Medical Center uses hydrostatically dilated saphenous venous graft to anastomose prominent segment V and VIII hepatic vein branches to the inferior vena cava. The University of Hong Kong Medical Centre includes the middle hepatic vein in every graft and anastomoses it to the recipient's middle or left hepatic vein. Kyoto University uses venous jump graft for anastomosing prominent middle hepatic vein branches to the inferior vena cava for recipients receiving a small-for-size graft or graft with dominant middle hepatic vein drainage. The Chang Gung Memorial Hospital adopts a flexible approach in inclusion of the middle hepatic vein in the graft depending on the donor size and the hepatic venous configuration of the right lobe graft. In summary, the criteria for inclusion and reconstruction of the middle hepatic vein vary. Further analysis is needed to confirm the importance of adequate drainage of the right anterior sector in right-lobe live-donor liver transplantation. PMID- 12589136 TI - Approach to anatomic variations of the graft portal vein in right lobe living donor liver transplantation. AB - Right lobe living-donor liver transplantation (LDLT) is often not attempted in donors with anomalous portal venous branching (APVB). The authors describe their experience with portal vein (PV) reconstruction in 17 cases of APVB in right lobe LDLT. From July 1997 to December 2001, 214 right liver LDLT were performed at the Asan Medical Center. Seventeen of the donors had APVB and successfully underwent right lobectomy. The APVB were type II (trifurcation) in nine cases, type III (independent posterior segmental branching from main PV trunk) in seven, and unclassified in one. All 17 donors and recipients are alive, with good liver function. In type II APVB, the donor PV branches were obtained with separate openings that were joined as a common orifice at the back table in two, with a discoid-patch single opening in four, and with one common opening in three. In type III APVB, the donor PV were divided with two openings in four and with a discoid-patch single opening in three. The discoid-patch defect in the remnant PV was repaired with a vein patchplasty in two donors and resected with end-to-end anastomosis in five. However, one donor developed portal vein thrombosis (PVT) that was managed successfully by re-exploration and insertion of a metallic vascular stent. Of the four type III APVB obtained with two separate PV openings, the first two liver grafts were each reconstructed as double PV anastomoses. One of them required re-exploration because of PVT. In the two succeeding cases, a Y graft interposition technique using a cryopreserved cadaveric iliac vein or the recipient's own portal confluence was successfully applied. To minimize the risk of PVT in donors with APVB, discoid-patch excision followed by repair with vein patchplasty or segmental resection should be avoided. Individual division of the PV branches creating two separate openings instead is recommended. To decrease the recipient's risk of PVT, interposition Y-graft venous reconstruction at the back table is superior to double PV anastomoses. PMID- 12589137 TI - Living-donor liver transplantation for high-urgency situations. AB - Patients in high-urgency situations for liver transplantation have a high mortality rate while on the waiting list for cadaveric grafts. In countries where cadaveric organ donation is scarce, the use of living-donor liver transplantation (LDLT) provides the only chance of survival for both pediatric and adult patients. It results in a satisfactory overall survival, from approximately 60% to 75% in pediatric patients and from 70% to 90% in adult patients. Patients who had early LDLT were shown to have a better survival outcome than those who waited for cadaveric organ donation, because a timely graft was available to them before they deteriorated further to multiple organ failure. Patients who were in high urgency situations for liver transplantation and opted for LDLT were also shown to have significantly better survival outcomes than those who did not opt for this procedure. Although left lobe liver graft can be used successfully in adult to-adult LDLT in high-urgency situations, there is a trend toward a more frequent use of the larger mass provided by the right lobe of the liver. Adequate venous drainage of the anterior segment of the right lobe liver graft is also considered essential for the favorable outcome of the recipients. Although donor morbidity has been reported to be low in LDLT and no donor death has been reported from Asian transplant centers, standardized definitions of morbidity and better methods for observing and measuring outcomes are necessary to understand and to potentially reduce morbidity. PMID- 12589138 TI - Living-donor liver transplantation for hepatocellular carcinoma. AB - In cadaveric liver transplantation, the Milan criteria have been accepted as the selection criteria for hepatocellular carcinoma (HCC) patients in considering organ allocation. However, the situation is different in living-donor liver transplantation (LDLT), in which the donor has a strong preference for altruism. The authors describe herein their experience with LDLT for HCC patients using their patient selection criteria. From February 1999 to March 2002, right lobe LDLT was performed in 56 patients with HCC. The authors' exclusion criteria included only those with extrahepatic metastasis or vascular invasion detected during preoperative evaluation. Thirty patients (54%) were in tumor, node, metastases stage IVa and 25 patients (45%) did not meet the Milan criteria at the time of LDLT. The follow-up period was 1 to 39 months (median, 11 months). The overall survival rates at 1 and 3 years were 73% and 55%, respectively, and the latter was significantly lower than that of adult right lobe LDLT without HCC (71% at 3 years). Fourteen patients died because of postoperative complications without tumor recurrence. Thirty-six patients survived without recurrence and six patients had recurrence. Among the six patients with recurrence, four had survived for 11 to 36 months after LDLT. In the analysis of patients who survived longer than 3 months after transplantation, 19 of 20 within the Milan criteria survived without recurrence. However, 15 of 20 patients beyond the criteria also survived without recurrence for 3 to 33 months (median, 12 months) and three of five patients with recurrence were alive for 11 to 36 months (median, 20 months). Histopathologic grading and microscopic portal venous invasion had significant negative impact on tumor recurrence. LDLT was an effective treatment for uncontrollable hepatocellular carcinoma. Because many patients who did not meet the Milan criteria survived without tumor recurrence after transplantation, different patient selection criteria are necessary in LDLT to save those with advanced HCC. PMID- 12589139 TI - Prophylaxis and treatment of recurrent hepatitis B after liver transplantation. AB - Chronic hepatitis B virus (HBV) infection is the leading cause of end-stage liver disease in Asia. The results of liver transplantation in these patients have significantly improved to a level comparable to those of other recipients as a result of the rapid evolution in the strategies of prevention and treatment of recurrent hepatitis B over the past decade. Hepatitis B immune globulin (HBIG) and lamivudine, either alone or in combination, are effective in preventing reinfection, but the most cost-effective dosing regimen with optimum efficacy without the prohibitive cost remains to be determined, an issue that is particularly relevant to liver transplant centers with serious financial constraints in Asia. The idea of active immunization is attractive, but the results have been conflicting to date. The newer nucleoside analogs appear most promising, and a combination of two or more antiviral agents is likely to represent the future strategy of choice in the prophylaxis and treatment of recurrent hepatitis B after liver transplantation. PMID- 12589140 TI - Safety of anti-hepatitis B core antibody-positive donors for living-donor liver transplantation. AB - Serologic evidence of resolved hepatitis B virus (HBV) infection (HBV surface antigen negative, anti-HBV core antibody [HBc] positive) in a liver donor can be regarded as an occult infection with episomal HBV in the liver. The purpose of this study was to evaluate the safety of anti-HBc-positive living donors. Between March 2001 and January 2002, 127 donors underwent hepatectomy for living-donor liver transplantation at Asan Medical Center. They were classified as members of an anti-HBc-positive group (n=50) or an anti-HBc-negative group (n=77). The two groups were subdivided into right lobectomy (n=86) and left lobectomy (n=34) groups to compare operative risk. Perioperative clinical profiles were compared by anti-HBc status and extent of donor hepatectomy. There were no statistical differences of preoperative liver function and liver steatosis between the anti HBc-positive and anti-HBc-negative groups. Operation time and blood loss did not show any differences between the hepatectomy-matched anti-HBc-positive and anti HBc-negative groups. Postoperative recovery of liver function, incidence of complication, and regeneration rate of the remnant liver after right lobectomy also did not show significant differences. The anti-HBc-positive group did not exhibit any adverse preoperative, intraoperative, or postoperative outcomes compared with the anti-HBc-negative group. This indicates that anti-HBc-positive donors can be assessed to have the same degree of risk for donor operation as anti-HBc-negative donors. PMID- 12589141 TI - Hepatitis B core antibody-positive grafts: recipient's risk. AB - The transmission of hepatitis B virus infection through hepatitis B core antibody (anti-HBc)-positive liver grafts in hepatitis B surface antigen (HBsAg)-negative recipients has been established. The mandatory use of immunosuppression in transplant patients favors reactivation of latent virus that may be present in grafts from HBsAg-negative anti-HBc-positive donors. With the persistent organ donor scarcity, the use of these grafts cannot be avoided, especially in urgent cases and in areas where the prevalence of the hepatitis B virus is high, as in Asia. The recognition of posttransplant de novo hepatitis B from core antibody positive liver donors has, therefore, led to modifications in graft allocation policies and the introduction of strategies for prophylaxis. The risk of developing this type of new-onset hepatitis B virus infection in liver transplant recipients and the various approaches to minimize this risk are reviewed. The peculiar implications of using core antibody-positive grafts in the context of living donor liver transplantation in Asia are discussed. PMID- 12589142 TI - Nonmelanoma skin cancer in organ transplant patients. AB - Nonmelanoma skin cancer (NMSC) is more frequent in immunocompromised patients, for example, patients with organ transplants. A number of studies have been published from different countries that present a similar picture of tumors in transplant patients. In addition, the behavior of these tumors is often more aggressive in this group of high-risk patients. The multitude of NMSC and precancerous lesions presents a clinical diagnostic and therapeutic challenge to the managing dermatologists. Technology is being developed to cope with the clinical diagnosis and medical adjunct treatment to broaden the therapeutic options. It is suggested that the optimal use of these new developments occurs if patients are seen in specialized clinics aimed at providing preventive measures, diagnosis, and treatment. PMID- 12589143 TI - Antibody-mediated accommodation of heart grafts expressing an incompatible carbohydrate antigen. AB - BACKGROUND: Accommodation in patients transplanted with ABO incompatible allografts describes a state in which antibodies are produced against the incompatible blood group carbohydrate antigen; however, the graft is not rejected. The present study describes an experimental model for antibody-mediated accommodation of organs expressing incompatible carbohydrate antigens. METHODS: The model includes alpha1,3galactosyltransferase knockout mice that lack the alpha-gal epitope (Galalpha1-3Galbeta1-4GlcNAc-R), transplanted heterotopically with wild-type (WT) hearts expressing this epitope. The mice are irradiated and receive memory anti-Gal B cells by adoptive transfer. Immunization of these mice with pig-kidney membranes induces the production of large amounts of anti-Gal, which binds specifically to alpha-gal epitopes. RESULTS: Under the described accommodation protocol, transplanted mice produce anti-Gal that binds to alpha gal epitopes on endothelial cells of the grafted WT heart; however, the WT hearts continued to function for months. Second WT hearts transplanted into accommodating, anti-Gal producing mice, were not rejected. Anti-Gal in accommodating mice was not cytolytic, whereas anti-Gal in rejecting mice readily induced complement-mediated lysis of cells expressing alpha-gal epitopes. In addition, accommodating mice displayed a preferential increase in the anti-Gal immunoglobulin (Ig)G2b subclass. CONCLUSIONS: The immune system may be manipulated to accommodate grafts expressing incompatible carbohydrate antigens by preferential production of noncytolytic anticarbohydrate antibodies. PMID- 12589144 TI - Adoptive immunotherapy against kidney targets in dog-leukocyte antigen-identical mixed hematopoietic canine chimeras. AB - BACKGROUND: Stable mixed-donor-host-hematopoietic chimerism can serve as a platform for adoptive immunotherapy. Infusions of donor lymphocytes (DLI) sensitized against hematopoietic cells converted mixed hematopoietic into full donor chimerism in dog-leukocyte antigen (DLA)-identical littermates. Whether sensitization against tissue of solid organs leads to organ-specific immunity that can be transferred by DLI was unknown and was investigated in these experiments using the kidney as target. METHODS: DLA-identical recipients with established stable mixed-donor-host-hematopoietic chimerism were used. In five pairs, hematopoietic stem-cell transplant (HSCT) donors were sensitized by kidney transplantation from the respective chimeras. In a second group, five HSCT donors received vaccinations that were generated from kidney cells of the respective mixed chimeras. Twenty-eight days after sensitization, DLI were administered to the mixed-hematopoietic chimeras. RESULTS: All HSCT donors rejected their kidney grafts from their mixed-chimeric recipients within 22 to 45 days. DLI caused no sustained graft-versus-kidney effects in the mixed-chimeric recipients. However, DLI donors sensitized by kidney transplantation converted 4 of 5 mixed chimeras into virtually complete (>95%) donor-type chimeras, compared with 1 of 5 mixed chimeras given DLI by vaccination from sensitized donors. CONCLUSION: Although DLA-identical kidney grafts from mixed-hematopoietic chimeras were readily rejected by their HSCT donors, subsequent transfusions of sensitized-donor lymphocytes into mixed chimeras converted mixed to all-donor chimerism but failed to induce graft-versus-kidney effects. Vaccination strategies in lieu of kidney grafts failed to convert mixed chimerism. PMID- 12589145 TI - Combined gene therapy with adenovirus vectors containing CTLA4Ig and CD40Ig prolongs survival of composite tissue allografts in rat model. AB - BACKGROUND: The blockade of costimulatory signal pathway by anti-CD40 ligand antibody or cytotoxic T lymphocyte antigen 4 immunoglobulin (CTLA4Ig) prolongs allograft survival in various vascularized organ transplantations. Because of the short half life of these agents, repeated administration of proteins is required to achieve significant graft survival. Furthermore, there is limited information regarding the effect of cosimulatory blockade on the survival of composite tissue allografts. Therefore, we examined the effect of adenovirus-mediated gene transfer of CTLA4Ig or CD40Ig gene or both in composite tissue allotransplantation. METHODS: The hind limbs removed from male ACI rats (RT1 ) were transplanted into female Lewis rats (RT1 ) heterotopically. The recombinant adenovirus carrying CTLA4Ig (AxCTLA4Ig) or CD40Ig (AxCD40Ig) was intravenously administered after limb transplantation. RESULTS: Limb allograft survival was significantly prolonged by either AxCTLA4Ig or AxCD40Ig treatment at 1 x 10 plaque forming unit (mean survival time [MST] of 39.4+/-6.0 and 13.0+/-2.9, respectively) compared with the adenovirus vector containing beta-galactosidase treated group (MST of 4.8+/-0.8). Combination of AxCTLA4Ig and AxCD40Ig led to significant prolongation of graft survival (MST of 49.2+/-6.6). Serum levels of CD40Ig were higher in rats treated with combination therapy than those treated with AxCD40Ig alone, whereas the serum levels of CTLA4Ig in rats treated with AxCTLA4Ig alone and AxCTLA4Ig and AxCD40Ig combined were very similar. CONCLUSION: This study indicates that an adenovirus-mediated gene therapy of CTLA4Ig or CD40Ig has a therapeutic potential for preventing rejection in composite tissue transplantation. Furthermore, a combination therapy of AxCTLA4Ig and AxCD40Ig was even more effective in preventing acute rejection and prolonging the survival of allografted limbs without apparent complication. PMID- 12589146 TI - Xenogeneic thymus transplantation in a pig-to-baboon model. AB - BACKGROUND: We have tested whether fetal porcine thymic tissue transplantation can lead to tolerance across a discordant (pig-to-baboon) xenogeneic barrier. METHODS: Six baboons underwent a conditioning regimen with thymectomy, splenectomy, and anti-monkey CD3 antibody conjugated to a diphtheria toxin binding site mutant (FN18-CRM9). Porcine fetal or neonatal thymic tissue was transplanted into three baboons. Three control baboons received either no transplanted pig tissue (n=1) or adult pig lymph node (n=2). Cellular responses and skin xenografts were used to test for tolerance. RESULTS: After T-cell depletion and thymic transplantation, recovery of thymus-dependent naive-type CD4 cells (CD4/CD45RA ) and in vitro xenogeneic hyporesponsiveness were observed. No sensitization of alpha-galactosyl antibody responses was observed. The thymic grafts survived up to 48 days. Porcine skin xenografts were performed in two of these animals with survival of 22 and 24 days. Only two of these animals were completely T-cell depleted, and both failed to recover thymus-dependent T cells (CD4/CD45RA ). In one animal, general in vitro hyporesponsiveness was observed, with subsequent death from infection. The second animal demonstrated delayed recovery of T cells and prolonged general hyporesponsiveness in vitro. Neither animal demonstrated prolongation of porcine skin grafts compared with allografts (both rejected by day 13). CONCLUSIONS: Porcine thymic tissue is able to induce xenogeneic hyporesponsiveness. More efficient thymic engraftment may allow this approach to induce xenograft tolerance. PMID- 12589147 TI - Allogeneic adrenocortical transplantation: glucocorticosteroid-independent immunomodulatory properties of adrenal cortex cells. AB - BACKGROUND: Hormone substitution for the treatment of adrenocortical insufficiency does not adequately substitute the physiologic circadian secretion of corticosteroids and leads to long-term sequelae and reduced quality of life. The lack of adaptation to physical and psychologic stress situations may lead to life-threatening Addison's crises. Allogeneic transplantation of adrenal cortex could offer an intriguing alternative. Adrenocortical grafts were demonstrated to proliferate and produce corticosteroids in physiologic concentrations after transplantation. METHODS: K -transgenic murine lymphocytes and allogeneic adrenal cortex cells were cocultured in mixed lymphocyte reactions to examine the alloimmune response; lymphocytes from T-cell receptor transgenic mice and normal mice, respectively, served as responder cells. The effects of corticosteroids secreted by adrenocortical cells were antagonized by the steroid receptor antagonist mifepristone, whereas the impact of cell-cell interactions was differentiated with transwell culture systems. RESULTS: Coculture of adrenal cortex cells in mixed lymphocyte reactions markedly suppressed lymphocyte proliferation. Transwell cultures demonstrated that adrenocortical cells exerted their effects by a soluble factor that was only partially antagonized by mifepristone. CONCLUSION: In vitro, the presence of adrenocortical cells potently suppressed allogeneic immune responses. This effect was not exclusively the result of the secretion of corticosteroids, indicating an additional immunomodulatory property of adrenocortical cells. PMID- 12589148 TI - Intraportal infusion of 99mtechnetium-macro-aggregrated albumin particles and hepatocytes in rabbits: assessment of shunting and portal hemodynamic changes. AB - BACKGROUND: Partial correction of metabolic liver disease by hepatocyte transplantation requires infusion of a large number of cells into the portal vein. Uncontrolled infusion of cells leads to extrahepatic shunting. Obstruction of the sinusoidal space may result in hemodynamic changes and impairment of liver function. METHODS: Catheters connected to a port were placed into the caudal mesenteric vein of rabbits. After injection of 99mtechnetium-macroaggregated albumin (99mTc-MAA) surrogates or 99mTc-MAA/hepatocyte (Hc) mixtures (1:125), shunting into the lung was scintigraphically monitored. Volume flow (mL/min) and maximum velocity of the portal vein were recorded by color-coded Doppler ultrasound during intraportal application of 2.5 x 10(7) MAA particles, 2.5 x 10(7) isolated hepatocytes, and saline solution without particles or cells. RESULTS: 99mTc-MAA particles (2.5 x 10(7)) or equivalent MAA/Hc mixtures were completely retained in the liver. With additional application of 2.5 x 10(7) particles, shunting into the lung was observed in two animals of the MAA group. All animals in the hepatocyte group have received 5 x 10(7) MAA/Hc mixtures, and three of these received 10(8) mixtures without shunting. Maximum velocity and volume flow increased with saline infusion. Hepatocyte suspended in the same volume blunted the increase observed in the control group, but parameters remained normal. Liver enzymes increased after hepatocyte application but returned to normal values within 5 days. CONCLUSIONS: Sinusoidal uptake capacity for hepatocyte or MAA particles varies at a wide range in normal rabbits. Scintigraphic monitoring of transplanted cells allows efficient monitoring of cell translocation into the lungs. No significant impairments of portal hemodynamics and liver function were detected. PMID- 12589149 TI - Long-term liver allograft survival induced by combined treatment with rAAV hCTLA4Ig gene transfer and low-dose FK506. AB - BACKGROUND: Recombinant adeno-associated virus vector (rAAV) is a promising vehicle for gene delivery, but few reports have documented its application in solid organ transplantation. In a rat orthotopic liver transplantation model, we investigated the efficacy of rAAV-mediated human cytotoxic T-lymphocyte associated antigen 4 and immunoglobulin G (hCTLA4Ig) gene transfer to induce long term allograft survival. METHODS: Dark Agouti and Lewis rats were used as donors and recipients, respectively, in six experimental groups: (a) syngeneic control, (b) no treatment, (c) rAAV-green fluorescent protein, (d) rAAV-hCTLA4Ig, (e) low dose FK506 for 7 days, and (f) rAAV-hCTLA4Ig and low-dose FK506 for 7 days. RESULTS: The liver allografts were rejected within 10 days when no treatment was given or rAAV-green fluorescent protein was delivered. rAAV-hCTLA4Ig transduction slightly prolonged the survival time to 11 days. Long-term survival was achieved using the combined treatment of rAAV-hCTLA4Ig and low-dose FK506, whereas grafts were rejected on day 33 in the low-dose FK506 group. A sustained hCTLA4 level in plasma was detected in the combined treatment group from day 5 to day 180. On postoperative day 5, combined treatment significantly decreased the interleukin-2 and interferon-gamma protein levels in the grafts and the number of infiltrating B, T, CD25+, CD4+, CD8+, and NK cells. CONCLUSIONS: This study shows that rAAV hCTLA4Ig gene transfer combined with low-dose FK506 can achieve long-term liver allograft survival. PMID- 12589150 TI - Synergistic effects of mycophenolate mofetil and losartan in a model of chronic cyclosporine nephropathy. AB - BACKGROUND: Combined treatments of mycophenolate mofetil (MMF) and losartan (LSRT) have synergistic effects on various renal diseases through their hemodynamic and anti-inflammatory effects. This study investigated whether MMF treatment is effective in inhibiting inflammatory processes in chronic cyclosporine A (CsA) nephrotoxicity, and whether combined treatment using MMF and LSRT affords superior protection compared with the respective monotherapies. METHODS: Rats on a low-salt diet were given vehicle (VH group, olive oil, 1 mg/kg per day), CsA (15 mg/kg per day), CsA and LSRT (CsA+LSRT group, 100 mg/L per day), CsA and MMF (CsA+MMF group; 40 mg/kg per day), or CsA, LSRT and MMF (CsA+LSRT MMF group). Control groups received each drug without CsA treatment. Renal function, histologic parameters (arteriolopathy, tubulointerstitial fibrosis, and inflammatory cell infiltration), and mediators of CsA-induced nephrotoxicity (angiotensin-II, osteopontin, and transforming growth factor [TGF] beta1) were studied. RESULTS: The CsA-treated rats showed decreased renal function and increased histologic parameters compared with the VH-treated rats. The CsA+MMF treatment significantly improved renal function and histopathologic parameters compared with the CsA group, and combined treatment with MMF and LSRT further improved those parameters compared with the CsA+LSRT and CsA+MMF groups. At a molecular level, increased expression of angiotensin II protein, osteopontin, and TGF-beta1 mRNAs in the CsA group were significantly decreased with MMF, and further decrease was observed with the combined treatment using MMF and LSRT. CONCLUSIONS: MMF treatment decreases CsA-induced nephrotoxicity, and combined treatment with LSRT has a synergistic effect in preventing chronic CsA nephrotoxicity. PMID- 12589151 TI - Bone disease after renal transplantation. AB - Bone disease is common after renal transplantation. The main syndromes are bone loss with a consequent fracture rate of 3% per year, osteonecrosis of the hip, and bone pain. The causes of disease include preexisting uremic osteodystrophy (hyperparathyroidism, aluminum osteomalacia, beta2-associated amyloidosis, and diabetic osteopathy), postoperative glucocorticoid therapy, poor renal function, and ongoing hyperparathyroidism, as the result of either autonomous transformation of the parathyroid gland or ongoing physiologic stimuli. Cyclosporine A treatment, hyperphosphaturia, and a pathogenic vitamin D allele have also been implicated. Bone loss is particularly pronounced during the first year after operation, amounting to up to 9% of bone mass. The clinical and biochemical picture is consistent with a high turnover bone disease, but histomorphometric studies do not completely support this. Principal prophylactic options include preoperative osteodystrophy prophylaxis; postoperative calcium, vitamin D, or calcitriol therapy; estrogen therapy for postmenopausal women; and parathyroidectomy for medically intractable hyperparathyroidism. Recently, prophylactic biphosphonate treatment has shown promise, but the exact indications for treatment remain to be determined. PMID- 12589152 TI - The isolation of Aspergillus fumigatus from respiratory tract specimens in heart transplant recipients is highly predictive of invasive aspergillosis. AB - BACKGROUND: Prompt recognition of invasive pulmonary aspergillosis (IPA) after heart transplantation is essential for achieving a successful outcome. However, the significance of the isolation of from respiratory specimens in heart transplant recipients is not established. METHODS: From 1990 to 1999, we analyzed first respiratory specimens with spp. growth from heart transplant patients in our institution. All specimens were cultured in both fungal and conventional media. Diagnosis of proven and probable IPA or colonization was made according to criteria of the Mycoses Study Group. RESULTS: During the 10-year study period, spp. was recovered from 30 episodes from 27 heart transplant recipients (incidence: 10.5%). Three episodes were classified as indeterminate and were included in the analysis in a double way, first considering them as true positives and afterward as true negatives, so ranges were obtained. After applying diagnostic criteria, 18 of 30 episodes were proven or probable IPA, and 9 episodes were colonizations. Accordingly, 7 to 8% of heart transplant recipients suffered an IPA, and the overall positive predictive value (PPV) was 60% to 70%. When analyzed by species, the PPV of recovering was 78% to 91%, whereas it was 0% for other species. The PPV increased to 88% to 100% when was recovered from a respiratory specimen other than sputum and decreased to 50% to 67% when it was recovered from sputum. The sensitivities of fungal and conventional media for the recovery of spp. were 95% to 100% and 33% to 38%, respectively. CONCLUSION: The isolation of from the respiratory tract of a heart transplant recipient is highly predictive of invasive aspergillosis. PMID- 12589153 TI - Clodronate treatment of established bone loss in cardiac recipients: a randomized study. AB - BACKGROUND: Bone loss has been reported as a complication after heart transplantation (HTx), and the increase in bone fractures is an effective problem. Treatment of osteoporosis has obtained mixed results. In this study we evaluate the effect of treatment with an oral bisphosphonate. METHODS: Sixty-four patients with low mineral density 6 months after HTx were randomized as follows: Group A received oral clodronate (1600 mg/day in two divided doses), and Group B received placebo. Every patient was also treated with 2000 mg/day of oral calcium carbonate. Bone mineral density (BMD) was measured by dual energy x-ray absorptiometry at the lumbar spine, 1/3 and 1/10 of the distal nondominant forearm before and after 12 months of treatment. Laboratory tests were performed at 3, 6, and 12 months of treatment. RESULTS: All patients demonstrated manifest bone loss 6 months after HTx compared with normal non-HTx controls (P=0.0001). After 1 year of clodronate therapy, BMD at the lumbar spine increased from 0.77+/ 1.4 g/cm(2) to 0.86 g/cm(2) (P=0.02). Laboratory tests did not show any significant variation, except for the bone isoenzyme of alkaline phosphatase, which showed a significant decrease after 1 year of treatment. The incidence of new fractures was 9.3% in the placebo group and 0% in the clodronate group. Therapy was well tolerated without impact on graft function. CONCLUSIONS: One year of clodronate therapy induced a significant increase in BMD at the lumbar spine in our HTx patients. Treatment was well tolerated without onset of new bone fractures. PMID- 12589154 TI - Platelet-derived growth factor receptor inhibition reduces allograft arteriosclerosis of heart and aorta in cholesterol-fed rabbits. AB - BACKGROUND: Crosstalk between pro-inflammatory cytokines and platelet-derived growth factor (PDGF) regulates smooth-muscle-cell proliferation in cardiac allograft arteriosclerosis. In this study, we tested the effect of STI 571, a novel orally active protein tyrosine kinase (PTK) inhibitor selective for PDGF receptor (PDGF-R) on transplant and accelerated arteriosclerosis in hypercholesterolemic rabbits. METHODS: Cardiac allografts were transplanted heterotopically from Dutch Belted to New Zealand White rabbits. A 0.5% cholesterol diet was begun 4 days before transplantation. Recipients received STI 571 5 mg/kg per day or vehicle intraperitoneally throughout the study period of 6 weeks. Cyclosporine A was given as background immunosuppression. RESULTS: In cardiac allografts of vehicle-treated rabbits, 76.2+/-2.1% of medium-sized arteries were affected by intimal thickening, and the percentage of arterial occlusion was 45.0+/-5.0%. Treatment with STI 571 reduced the incidence of affected medium-sized arteries to 41.2+/-8.1% (P <0.05) and the arterial occlusion to 27.6+/-5.0% ( P<0.05). In addition, we observed that STI 571 treatment reduced intimal lesion formation in proximal ascending aorta of transplanted hearts from 72.3+/-19.9 to 12.7+/-1.9 microm ( P<0.05). Our results also show that STI 571 significantly inhibited accelerated arteriosclerosis in medium-sized arteries of recipients' own hearts. CONCLUSIONS: The results of the present study suggest that PDGF-R activation may regulate the development of transplant and accelerated arteriosclerosis in hypercholesterolemic rabbits. Thus, PTK inhibitors may provide new strategies for prevention of these fibroproliferative vascular disorders. PMID- 12589155 TI - Successful toxoplasmosis prophylaxis after orthotopic cardiac transplantation with trimethoprim-sulfamethoxazole. AB - BACKGROUND: The efficacy of trimethoprim-sulfamethoxazole (TMP/SMX) in the prevention of toxoplasmosis after orthotopic cardiac transplantation has been the subject of some controversy, with many transplant groups preferring to use the combination of pyrimethamine and sulfadiazine. Although effective, this latter regimen does not offer equal protection against other pathogens, such as or. To assess the value of TMP/SMX, we reviewed the experience in our heart transplant patients, all of whom received TMP/SMX (160/800 mg) three times weekly for approximately 8 months after transplantation. METHODS: We report on 417 orthotopic cardiac transplants during a 17-year period. We have 100% one-year patient follow-up after transplantation. Data was collected on pretransplantation donor and recipient anti- serology, immunosuppression, allograft rejection, survival, yearly posttransplantation anti- serology, development of acute toxoplasmosis, and the occurrence of other infections. RESULTS: In this cohort, acute toxoplasmosis developed after transplantation in one case (0.2%). Among the highest risk patients (D+R-) who were treated for at least one episode of rejection, the risk of acute toxoplasmosis was 5% (1 of 22 patients). No change in survival was found between the different anti- IgG serogroups (D-R-, D-R+, D+R , or D+R+). Anti- IgG seroconversion occurred in eight -seronegative recipients after transplantation; all patients, except the case already noted, were asymptomatic and required no specific anti- therapy. No cases of, or infections were identified. Five proven and two suspected cases of pneumonia were found (only 2 of these 7 patients were receiving TMP/SMX at the time of pneumonia diagnosis). CONCLUSIONS: These data demonstrate that TMP/SMX prophylaxis (160/800 mg) three times per week is effective prophylaxis after orthotopic cardiac transplantation and has prophylactic benefits against other posttransplantation opportunistic pathogens. PMID- 12589156 TI - The presence of glomerular sclerosis at time zero has a significant impact on function after cadaveric renal transplantation. AB - INTRODUCTION: The aim was to determine the impact of donor glomerulosclerosis on allograft outcome. METHODS: The percentage of glomerular sclerosis (%GS) was calculated in protocol biopsies taken at engraftment. Clinical variables were obtained from the Welsh Transplantation Research Group (WTRG) database. RESULTS: Of 210 allografts, 129 showed %GS=0, but 81 kidneys showed %GS between 1 and 60. Patients with %GS=0 had the highest glomerular filtration rate (GFR) at 1 year (62.0 mL/min) and the slowest deterioration of function (-3.8 mL/min per year). Patients with %GS greater than 20 had the lowest GFR at 1 year (36.0 mL/min) and the steepest rate of deterioration (-9.0 mL/min per year). The %GS of 10 alone can reduce GFR at 4 years by 8 mL/min, a similar reduction to a single rejection episode or an increase in donor age of 30 years. Actuarial 5-year graft survival for %GS=0 was 80%, and for %GS greater than 20 was 35% ( P=0.04). CONCLUSION: The findings indicate that a biopsy taken at procurement will provide information for the most appropriate allocation of a kidney. PMID- 12589157 TI - Highly urgent liver transplantation: possible impact of donor-recipient ABO matching on the outcome after transplantation. AB - BACKGROUND: Survival after liver transplantation for fulminant hepatic failure has been reported to be less favorable than survival for patients with chronic liver diseases. METHODS: We have studied all patients (n=229) undergoing highly urgent liver transplantation from 1990 to 2001 in the Nordic countries. The impact of patient and donor characteristics, with emphasis on donor-recipient ABO matching (identical, compatible, incompatible), has been studied. RESULTS: One year and 3-year patient survival rates were 73% and 70% for the total period and 86% and 78% for the last 4-year period. Patients receiving an ABO-compatible liver allograft had significantly lower patient survival rates than those receiving an ABO-identical donor organ (1-year patient survival rates 66% of vs. 79%, P=0.03). Graft survival rates varied less (1-year graft survival rates of 64% vs. 74%, P=0.09). Patients receiving an ABO-incompatible liver allograft had patient survival rates of 70% at 1 year and 60% at 3 years but low graft survival rates (40% and 30% at 1 and 3 years). In a multiple regression analysis, significant independent predictors of poor patient survival were early year of transplantation, ABO-compatible donor, high donor age, and waiting time more than 3 days and less than 9 days. CONCLUSION: Survival after highly urgent liver transplantation has improved and is comparable to that observed in patients receiving a liver allograft because of chronic liver disease. Patients receiving an ABO-identical donor organ had significantly higher patient survival rates compared with those receiving an ABO-compatible donor liver. PMID- 12589158 TI - Rabbit antithymocyte globulin as induction immunotherapy in pediatric heart transplantation. AB - BACKGROUND: There is little published data on the use of antithymocyte globulins in children. This retrospective study describes the use of Thymoglobulin (Imtix, SangStat, Lyon, France) in pediatric cardiac transplantation over a 13-year period in a single center that adjusted the dose of Thymoglobulin according to platelet count monitoring and examines the short-term hematological effects as well as longer-term outcomes. METHODS: Data for all children who received a heart transplant at the Hopital Cardiologique at Lyon from 1984 to 2001 and who were given Thymoglobulin as part of their immunosuppressive protocol were extracted. The dose of Thymoglobulin given depended on baseline platelet count and was 2, 1.5, or 1 mg/kg per day over 5 days for the following platelet count groups: greater than 150,000/mm (normal group), 100 to 150,000/mm (mild thrombocytopenia group), and 50 to 100,000/mm (moderate thrombocytopenia group). RESULTS: Thirty children of median age 14.2 years were given a median cumulative dose of Thymoglobulin of 8 mg/kg per patient; the moderate thrombocytopenia subgroup was given significantly less (6.4 mg/kg) ( P=0.032). Immediate tolerability of Thymoglobulin was good, with no cases of first-dose syndrome, anaphylaxis, or serum sickness. The platelet count decreased at the start of therapy, but recovered after discontinuation, and did not give rise to clinical concern. Patients were followed up for a median of 6.3 years (7 days-15.5 years); actuarial survival was 90%, 86%, and 74.5%, respectively, at 1, 5, and 10 years. In the first year, 50% of patients suffered an episode of rejection. The overall incidence of infection in the month following transplantation was 40%. One lymphoma occurred at 5 months. CONCLUSIONS: The use of Thymoglobulin in pediatric heart-transplant patients as part of an immunosuppressive protocol, with dose adjustment according to platelet levels, has been shown to be effective in terms of rejection rate and patient survival and safe in terms of the incidence of infections and malignancy. PMID- 12589159 TI - Hepatic venoplasty in living-donor liver transplantation using right lobe graft with middle hepatic vein. AB - Inclusion of the middle hepatic vein in a right lobe graft from a living-donor may improve venous drainage and avoid graft dysfunction, but reconstruction of the middle hepatic vein is technically difficult. We developed a hepatic venoplasty technique, which was applied in eight consecutive right lobe liver transplantations. The right and middle hepatic veins of the graft were joined together to form a triangular cuff for a single anastomosis to the recipient's inferior vena cava. Hepatic venoplasty was successful in all cases, and no interposition graft was required. Venovenous bypass was not used. All grafts showed immediate function, and no hepatic venous outflow obstruction was observed. There was no reoperation and the graft survival rate was 100%. This hepatic venoplasty technique can be applied systemically as a standard one in right lobe liver graft with the middle hepatic vein to simplify the recipient hepatectomy and to obviate venous outflow obstruction. PMID- 12589160 TI - Multivariate analysis of donor risk factors for graft survival in kidney transplantation. AB - BACKGROUND: The results of the transplantation of marginal donor kidneys remain controversial. This study aimed to investigate the impact of donor risk factors as predictors of kidney-graft outcome. METHODS: Allograft failure risk factors were studied in 7,209 cadaveric kidney-transplant recipients reporting to the Etablissement francais des Greffes (EfG) from 1996 to 2000, of which 544 (7.6%) were from donors aged over 60. Both univariate and multivariate analysis were used to assess the effect of donor risk factors and were stratified according to recipient age. RESULTS: Overall graft survival was 91.1% (95% confidence interval [CI] 90.5-91.8) at 1 year, 88.6% (95% CI 87.8-89.4) at 2 years, and 85.6% (95% CI 84.6-86.6) at 3 years posttransplant. Univariate analysis of risk factors showed a significant reduction of graft survival in recipients transplanted with kidneys coming from donors older than 60 years, donors with a history of hypertension, a cerebrovascular cause of death, and a preharvesting serum creatinine greater than 150 micromol/L. Multivariate analysis revealed significantly higher failure rate associated with cerebrovascular cause of death (RR=1.2, P=0.02), history of hypertension (RR=1.2, P=0.04), and elevated serum creatinine (RR=1.3, P=0.03), whereas donor age greater than 60 years was not found as an independent risk factor. CONCLUSIONS: Our results suggest that cerebrovascular cause of death, history of hypertension, and elevated creatinine are significant independent donor risk factors for graft survival, whereas donor age is a statistically significant, but dependent, risk factor. This result is important for the design of allocation and transplantation strategies for kidneys procured in elderly donors. PMID- 12589161 TI - A short course of cyclosporine immunosuppression inhibits rejection but not tolerance of rat liver allografts. AB - BACKGROUND: Orthotopic liver transplants in many animal models are spontaneously accepted without requiring immunosuppression. Liver transplant acceptance is associated with early immune activation, and immunosuppressive drugs such as methylprednisolone inhibit acceptance. We investigated whether cyclosporine (CsA) inhibits rat liver transplant acceptance. We also examined the effects of CsA on infiltration and cytokine gene expression. METHODS: Orthotopic liver transplantation was performed in the PVG donor to Dark Agouti recipient rat strain combination, which accepts the graft (tolerance; TOL), and in the PVG-to Lewis combination, which rejects the graft in 9 to 16 days (rejection; REJ). CsA (1.5 mg/kg per day subcutaneously) was given to recipients for 5 days, starting from the day of transplantation to day 4 or from day 3 to day 7. In a separate experiment, transplanted livers were collected at days 1, 3, 5, and 7 after transplantation and examined for infiltration by immunohistochemistry and for expression of interleukin (IL)-2, IL-4, IL-10, and interferon-gamma mRNA by quantitative reverse transcriptase-polymerase chain reaction. RESULTS: Both early and delayed treatment with CsA significantly increased survival in the REJ strain combination, with a median survival time of 81 days and more than 100 days, respectively, compared with 13 days in the untreated group. Neither treatment affected survival of TOL animals, and all TOL groups had a median survival time of more than 100 days. Delayed treatment did not reduce survival; more animals survived for greater than 100 days after delayed treatment, although this did not reach significance ( P=0.08). T-cell infiltrate was inhibited in CsA-treated TOL animals compared with untreated animals at all times after treatment, whereas CD25 cells were only inhibited on day 3. CsA treatment of TOL grafts markedly reduced expression of IL-2, IL-4, and interferon-gamma compared with untreated recipients. CONCLUSIONS: CsA did not significantly inhibit liver transplant acceptance and allowed some activation of T cells and CD25 expression but almost completely inhibited IL-2 and IL-4, which are required for survival of activated T cells. PMID- 12589162 TI - Inhibition of complement factor C5 protects against renal ischemia-reperfusion injury: inhibition of late apoptosis and inflammation. AB - BACKGROUND: Complement has been implicated in the pathophysiology of renal ischemia-reperfusion (I/R) injury. However, the mechanism underlying complement mediated renal I/R injury is thus far unknown. To investigate the involvement of complement in I/R injury, we studied the activation and deposition of complement in a murine model of renal I/R injury. Furthermore, we examined the effect of inhibition of complement-factor C5 on renal I/R injury. METHODS: Mice were subjected to 45 min of unilateral ischemia and subsequent contralateral nephrectomy and reperfusion for 2, 12, or 24 hr. Mice were control treated or treated with BB5.1, a monoclonal antibody that prevents cleavage of complement factor C5, thereby preventing C5a generation and formation of the membrane attack complex (MAC). RESULTS: Renal I/R induced extensive deposition of C3 early after reperfusion, whereas C6 and C9 deposition (MAC formation) occurred relatively late. I/R-induced complement deposition was mainly localized to tubular epithelium. Treatment with BB5.1 totally prevented MAC formation but also reduced C3 deposition. Inhibition of C5 strongly inhibited late inflammation, as measured by neutrophil influx and induction of the murine CXC chemokines macrophage inflammatory protein-2, KC, and lipopolysaccharide-induced CXC chemokine. Anti-C5 treatment furthermore abrogated late I/R-induced apoptosis, whereas early apoptosis was not affected. Moreover, BB5.1 treatment significantly protected against I/R-induced renal dysfunction. CONCLUSIONS: Renal I/R is followed by activation of the complement system and intrarenal deposition of C3 and MAC. Complement activation plays a crucial role in the regulation of inflammation and late apoptosis. Complement inhibition, by preventing C5 activation, abrogates late apoptosis and inflammation, being strongly protective against renal function loss. PMID- 12589163 TI - Dendritic cells exposed to nacystelyn are refractory to maturation and promote the emergence of alloreactive regulatory t cells. AB - BACKGROUND: Drugs blocking dendritic cell (DC) maturation might be useful in transplantation by inhibiting the induction of primary alloimmune responses and promoting the emergence of regulatory T lymphocytes (Treg). We investigated the effects of Nacystelyn (NAL), an N-acetyl-L-cysteine derivative, on human DCs, paying attention to the T-cell responses elicited by NAL-treated DCs in vitro. METHODS: Lipopolysaccharide (LPS) was used to induce the maturation of DCs naturally present in blood or generated from human monocytes cultured in interleukin-4 and granulocyte-macrophage colony-stimulating activity. We first analyzed the consequences of NAL on cytokine production and expression of major histocompatibility complex class II and costimulatory molecules. Monocyte-derived DCs were then used as stimulators in mixed leukocyte cultures with naive CD4 T cells. Cytokine levels were measured in culture supernatants; the phenotype of T cells and their capacity to inhibit the proliferation of third-party T-cell responders was determined at the end of the culture. RESULTS: NAL proved to be a potent inhibitor of DC maturation in whole blood experiments and on monocyte derived DCs. Alloreactive T cells stimulated with DCs pretreated with LPS in the presence of NAL produced much less interferon-gamma but similar levels of interleukin-13 compared with DCs treated with LPS alone. Immature DCs induced Treg, which was not observed with mature DCs. DCs cultured with LPS in the presence of NAL were as efficient as immature DCs to generate alloreactive T cells with regulatory activity. CONCLUSIONS: NAL is a potent inhibitor of DC maturation, which might be useful to promote allograft acceptance by inducing the differentiation of allospecific Treg. PMID- 12589164 TI - Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. AB - BACKGROUND: Marrow stromal cells (MSC) can differentiate into multiple mesenchymal tissues. To assess the feasibility of human MSC transplantation, we evaluated the in vitro immunogenicity of MSC and their ability to function as alloantigen presenting cells (APC). METHODS: Human MSC were derived and used in mixed cell cultures with allogeneic peripheral blood mononuclear cells (PBMC). Expression of immunoregulatory molecules on MSC was analyzed by flow cytometry. An MSC-associated suppressive activity was analyzed using cell-proliferation assays and enzyme-linked immunoassays. RESULTS: MSC failed to elicit a proliferative response when cocultured with allogeneic PBMC, despite provision of a costimulatory signal delivered by an anti-CD28 antibody and pretreatment of MSC with gamma-interferon. MSC express major histocompatibility complex (MHC) class I and lymphocyte function-associated antigen (LFA)-3 antigens constitutively and MHC class II and intercellular adhesion molecule (ICAM)-1 antigens upon gamma interferon treatment but do not express CD80, CD86, or CD40 costimulatory molecules. MSC actively suppressed proliferation of responder PBMC stimulated by third-party allogeneic PBMC as well as T cells stimulated by anti-CD3 and anti CD28 antibodies. Separation of MSC and PBMC by a semipermeable membrane did not abrogate the suppression. The suppressive activity could not be accounted for by MSC production of interleukin-10, transforming growth factor-beta1, or prostaglandin E2, nor by tryptophan depletion of the culture medium. CONCLUSIONS: Human MSC fail to stimulate allogeneic PBMC or T-cell proliferation in mixed cell cultures. Unlike other nonprofessional APC, this failure of function is not reversed by provision of CD28-mediated costimulation nor gamma-interferon pretreatment. Rather, MSC actively inhibit T-cell proliferation, suggesting that allogeneic MSC transplantation might be accomplished without the need for significant host immunosuppression. PMID- 12589165 TI - Skew in T-cell receptor usage and clonal T-cell expansion in patients with chronic rejection of transplanted kidneys. AB - BACKGROUND: It is not known whether alloreactive T cells are involved in chronic rejection of transplanted kidneys. The aim of the present study was to determine the involvement of T cells in the chronic graft rejection. METHODS: T-cell receptor (TCR) variable region alpha-chain and TCR variable region beta-chain repertoires were analyzed in peripheral blood mononuclear cells. T-cell clonalities were analyzed by complementarity-determining region 3 size spectratyping. RESULTS: A significant increase in the frequencies of one or more TCR variable region alpha-chain and TCR variable region beta-chain segments was detected in 13 and 15 of the 24 kidney transplant recipients, respectively. The extent of the skew in the TCR usage was correlated with the levels of clonal T cell expansion, indicating that the clonally expanding T cells were responsible for the skew in the TCR usage. The levels of the skew in the TCR usage and clonal T-cell expansion were significantly greater in the recipients with a graft failure than in those with a stable graft function ( P=0.0081 and P=0.012, respectively). These results indicate that the clonally expanding T cells in the periphery may be related to graft rejection. The percent increase in the serum creatinine levels, which reflected the deterioration of the kidney functions, was significantly higher in the recipients who showed high levels of clonal T-cell expansion than in those who did not ( P=0.021). CONCLUSIONS: The results demonstrate that clonal T-cell expansion in the periphery has a negative impact on the long-term graft functions, and that analysis of the clonal T-cell expansion in peripheral blood mononuclear cells provide significant information on the fate of the transplanted kidney. PMID- 12589166 TI - Bone marrow-derived immature dendritic cells prime in vivo alloreactive T cells for interleukin-4-dependent rejection of major histocompatibility complex class II antigen-disparate cardiac allograft. AB - BACKGROUND: Dendritic cells (DC) at the immature state express low levels of major histocompatibility complex and costimulatory molecules and are poor stimulators of primary T-cell response in vitro. Injection of immature bone marrow-derived DC, however, was shown to prime in vivo alloreactive CD4 T lymphocytes toward type 2 cytokine-producing cells in the absence of CD8 T-cell activation. METHODS: We undertook the present study to determine whether Th2 immunization by immature DC could lead to allograft rejection. We first analyzed, in the major histocompatibility complex class II antigen-disparate B6-anti-bm12 combination, the capacity of immature DC to regulate the activity of alloreactive CD4 T cells. We then determined, in this model of weak antigenicity, whether injection of bm12 DC in B6 recipients before transplantation could modify the survival of vascularized bm12 cardiac allografts. RESULTS: We confirmed that in vitro immature DC are poor stimulators of T-cell alloresponse. However, when given in vivo, immature bm12 DC primed anti-bm12 T cells for the production of interleukin (IL)-4. Moreover, they induced the acute rejection of bm12 cardiac allograft. The process of rejection was dependent on IL-4 because immunization of IL-4-deficient mice did not trigger rejection. CONCLUSIONS: Allogeneic immature DC generated with granulocyte-macrophage colony-stimulating factor are potent stimulators of primary alloreactive response in vivo and prime for transplant rejection. Our results indicate that strategies based on immature DC for the induction of transplantation tolerance should be considered with caution. PMID- 12589167 TI - Simultaneous kidney-pancreas transplantation in a patient with small lymphocytic lymphoma. AB - BACKGROUND: Experience with organ transplantation in patients with indolent lymphoma is limited, and it is unknown how the natural history of the disease is altered by chronic immunosuppressive therapy. METHODS: A patient with type 1 diabetes and renal failure who underwent simultaneous kidney-pancreas transplantation was found to have stage IV small lymphocytic lymphoma at the time of transplantation. He received quadruple immunosuppressive therapy using interleukin (IL)-2 receptor antibody, tacrolimus, mycophenolate mofetil, and prednisone. RESULTS: Patient is doing well 3 years posttransplant with excellent graft function of both the kidney and pancreas without any evidence of progression of the disease. CONCLUSION: Indolent lymphoma should not be considered an absolute contraindication to organ transplantation. PMID- 12589168 TI - Cholangitis caused by Roux-en-Y hepaticojejunostomy obstruction by a biliary stone after liver transplantation. AB - Biliary stones may occur after liver transplantation, mostly in relation with biliary strictures. We describe an unusual case of biliary stone developing 18 years after liver transplantation and biliary-enteric anastomosis for biliary atresia, responsible for an acute cholangitis caused by obstruction of the jejunal loop. Surgical exploration allowed the removal of the stone and did not reveal any enteric stricture. Both biliary and enteric anastomosis were patent. An hepatobiliary scintigraphy performed 2 weeks after surgery demonstrated a delayed bile excretion through the jejunal limb and a focal accumulation of the tracer at the level where the stone was found. In conclusion, biliary stone formation could develop after liver transplantation and biliary-enteric anastomosis, in the absence of mechanical stricture, secondarily to bile stasis caused by functional disorder of the efferent jejunal loop. Hepatobiliary scintigraphy plays a central role in the diagnosis and comprehension of such a disorder. PMID- 12589169 TI - Differential immunogenicity of HLA mismatches: HLA-A2 versus HLA-A28. AB - The immunogenicity of human leukocyte antigen (HLA)-A2 versus HLA-A28 was analyzed by antibody production, cytotoxic T-lymphocyte (CTL) induction, and graft survival. We observed that an HLA-A2 mismatched child in HLA-A28 women leads to HLA-A2 specific antibodies in 32% of the women (n=31), whereas in the case of an HLA-A28 child and HLA-A2 women (n=30), no HLA-A28 specific antibodies were found ( P<0.002). Also, the CTL precursor frequencies were significantly lower against HLA-A28 compared with CTLp frequencies against HLA-A2 ( P=0.012). Finally, the kidney graft survival was slightly better in HLA-A2 positive recipients transplanted with HLA-A28 mismatches. We can conclude that single HLA A28 mismatches are less immunogenic in HLA-A2 individuals compared with single HLA-A2 mismatches in HLA-A28 individuals, which is probably because the mismatched epitopes on the HLA-A2 molecule are unique epitopes, whereas the mismatched epitopes on HLA-A28 are shared by other HLA-A and HLA-B molecules. PMID- 12589170 TI - Evaluation of posttransplantation soluble CD30 for diagnosis of acute renal allograft rejection. AB - BACKGROUND: Posttransplantation measurement of soluble CD30 (sCD30) may be useful for identifying kidney graft recipients at risk of impending graft rejection in the early posttransplantation period. METHODS: We measured plasma sCD30 levels and evaluated the levels in relation to the diagnosis of rejection. RESULTS: Receiver operating characteristic curves demonstrated that on posttransplantation days 3 to 5, sCD30 allowed a differentiation of recipients who subsequently developed acute allograft rejection (n=25) from recipients with an uncomplicated course (n=20, P<0.0001) (area under the receiver operating characteristic curve 0.96, specificity 100%, sensitivity 88%) and recipients with acute tubular necrosis in the absence of rejection (n=11, P=0.001) (area under the receiver operating characteristic curve 0.85, specificity 91%, sensitivity 72%). CONCLUSIONS: sCD30 measured on posttransplantation days 3 to 5 offers a noninvasive means for differentiating patients with impending acute allograft rejection from patients with an uncomplicated course or with acute tubular necrosis. PMID- 12589171 TI - Calcium-channel blockers and end-stage renal disease: pharmacokinetic and pharmacodynamic considerations. AB - PURPOSE OF REVIEW: To characterize the pharmacokinetics and pharmacodynamics of the different calcium-channel blockers. RECENT FINDINGS: Calcium-channel blockers have been in use for some time in the end-stage renal disease population. Their primary use has been as antihypertensive and antianginal therapies. In this regard, they are effective agents. Recently, it has been noted that dialysis related hypotension occurs less frequently in calcium-channel blocker treated patients. Also, access patency and overall patient survival are improved with calcium-channel blocker therapy. SUMMARY: Calcium-channel blockers are useful agents for the control of hypertension in end-stage renal disease patients and appear to favorably influence survival in this population. Calcium-channel blockers are not dialyzable and their pharmacokinetics do not substantially change with renal failure therefore they do not require dose adjustment based on level of renal function. Too few studies exist to determine if individual calcium channel blockers differ in their effects. Prospective, randomized, controlled clinical trials are needed in the end-stage renal disease population to better understand the role of calcium-channel blockers in the excess cardiovascular disease burden of this population. PMID- 12589172 TI - Anaemia, cardiovascular disease and kidney disease: integrating new knowledge in 2002. AB - PURPOSE OF REVIEW: The present review assesses recent publications, from 2001 until the present, which address the relationship between cardiovascular disease (CVD) and anaemia in patients with chronic kidney disease. RECENT FINDINGS: Insights from the recently published basic science literature have helped to place findings from clinical studies into a new context, and thereby assist us to understand and further explore the complex relationship between haemoglobin level and survival in chronic kidney disease. The effects of erythropoietin molecules and the presence of receptors in vascular endothelium, myocardium and other tissues are described. Both observational and interventional clinical studies are examined, and limitations in the methodology and statistical analysis of clinical studies are emphasized, but are given context within the body of literature preceding the past year's publications. SUMMARY: Data suggest that development of CVD in patients with kidney disease is multifactorial. Several factors associated with CVD are also associated with anaemia, thereby making causal arguments for the role of anaemia in CVD and survival difficult. Arguments are made for the importance of prevention of anaemia and of individualizing therapeutic goals for its treatment. Well designed prospective studies with both CVD events and mortality as outcomes, and with enrollment beginning before the start of dialysis, are essential if we are to determine the optimal therapeutic strategies. PMID- 12589173 TI - Anaemia as a risk factor for the progression of chronic kidney disease. AB - PURPOSE OF REVIEW: About a dozen controlled clinical trials examined the effect of anaemia correction on the progression of chronic kidney disease. None of these studies fulfilled the stringent criteria of a randomized controlled trial as suggested by the CONSORT statement, yet evidence emerged that anaemia sustains mitogenic and fibrogenic stimuli by lowering local partial oxygen tension. This review addresses the question of why and how anaemia could possibly enhance the progression of chronic kidney disease, and summarizes relevant clinical trials. RECENT FINDINGS: The discovery of hypoxia-inducible factor, a transcription factor stabilized under hypoxic conditions, with DNA-binding properties towards about 50 target genes including erythropoietin, has largely encouraged the hypothesis that tissue hypoxia may serve as another common mechanism for the progression of chronic kidney disease besides hypertension or proteinuria. In addition, anaemia-mediated alterations of renal sympathetic nerve activity and anaemia-related increments of oxidative stress may contribute to a progressive nephron loss. Conclusive evidence from clinical trials is scarce. SUMMARY: Pathophysiological concepts suggest some impact of anaemia on the progression of chronic kidney disease. The urge for more sound clinical intervention trials is met by the ongoing ECAP study (Effect of early Correction of Anaemia on the Progression of chronic kidney disease). PMID- 12589174 TI - Iron metabolism in end stage renal failure: rationale for re-evaluation of parenteral iron therapy. AB - PURPOSE OF REVIEW: In this article we will examine the basis for using chronic high dose parenteral iron therapy in dialysis patients. RECENT FINDINGS: There are increasing data that dialysis patients fare better in many respects if they have higher hematocrit values although the real optimal hematocrit has not been defined. There is an increasing tendency to use parenteral iron to achieve this goal. SUMMARY: Although parenteral iron achieves seemingly favourable short results, there are no data for its safety in the long term. On the contrary, there are reasons to suggest possible iron overload with chronic use. PMID- 12589175 TI - Aldo is back: recent advances and unresolved controversies in hyperaldosteronism. AB - PURPOSE OF REVIEW: Hyperaldosteronism in its various forms is a recognized secondary cause of hypertension, yet the frequency of these disorders and the appropriate evaluation of suspected patients remain controversial. This review will summarize recent literature concerning the frequency of hyperaldosteronism in the hypertensive population, insight from uncommon forms of hyperaldosteronism, and new developments in the diagnosis and treatment of this condition. RECENT FINDINGS: Several series report that around 10% of hypertensive patients have some form of hyperaldosteronism, but aldosterone-producing adenomas are rare. Diagnostic criteria for idiopathic hyperaldosteronism remain controversial, as is the wisdom of widespread screening. Patients with even mild hyperaldosteronism, however, which could be a continuum with low-renin hypertension, may respond exceptionally well to mineralocorticoid antagonism. Eplerenone, a new mineralocorticoid receptor antagonist without antiandrogen side effects, has been an effective antihypertensive in clinical trials and appears to be particularly suitable for low-renin hypertensives. Accumulating evidence suggests that aldosterone excess is cardiotoxic and nephrotoxic, suggesting that mineralocorticoid blockade has specific benefits beyond blood pressure reduction. For patients with severe, confirmed hyperaldosteronism, selective adrenal vein sampling is the only reliable method for determining the source of the aldosterone. SUMMARY: Hyperaldosteronism, when defined with liberal criteria, could account for a substantial portion of hypertension. Few of these patients will harbor adrenal adenomas, but those with severe hypertension and hypokalemia often require adrenal vein sampling to direct surgery. With more precise diagnostic strategies, better treatments, and evolving evidence of pathological consequences of aldosterone excess, subtle disorders of aldosterone excess demand precise definition and specific treatment. PMID- 12589176 TI - Aldosterone in renal disease. AB - PURPOSE OF REVIEW: Interruption of the renin-angiotensin-aldosterone system, chiefly with angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers, has yielded beneficial results in retarding injury and progression in numerous intrinsic renal diseases. The renoprotection offered by these agents is incomplete and far from optimal. Studying mediators of progression other than angiotensin II is therefore extremely important. The emerging role of aldosterone in progression of renal disease and the utility of its antagonism is discussed here. RECENT FINDINGS: The experimental evidence linking aldosterone to renal disease is discussed. The exciting results from clinical studies employing mineralocorticoid receptor blockers are also described. SUMMARY: Aldosterone antagonism offers additional antiproteinuric benefits to those achieved with angiotensin-converting enzyme inhibition. Long-term trials addressing effectiveness and safety, especially in regards to hyperkalemia, are greatly needed. PMID- 12589177 TI - Recent advances in intracellular signalling in hypertension. AB - PURPOSE OF REVIEW: Transmission of external signals from the cell surface to the internal cellular environment occurs via tightly controlled complex transduction pathways. Alterations in these highly regulated signalling cascades in vascular smooth cells may play a fundamental role in the structural, mechanical and functional abnormalities that underlie vascular pathological processes in hypertension. The present review focuses on recent developments relating to two novel signalling pathways: angiotensin II signalling through tyrosine kinases; and oxidative stress and redox-dependent signal transduction. These pathways are emerging as critical mediators of hypertensive vascular disease because they influence multiple cellular responses that are involved in structural remodelling, vascular inflammation and altered tone. RECENT FINDINGS: A recent advance in the field of angiotensin II signalling was the demonstration that, in addition to its vasoconstrictor properties, angiotensin II has potent mitogenic like and proinflammatory-like characteristics. These actions are mediated through phosphorylation of both nonreceptor tyrosine kinases and receptor tyrosine kinases. It is also becoming increasingly apparent that many signalling events that underlie abnormal vascular function in hypertension are influenced by changes in intracellular redox status. In particular, increased bioavailability of reactive oxygen species (oxidative stress) stimulates growth-signalling pathways, induces expression of proinflammatory genes, alters contraction excitation coupling and impairs endothelial function. SUMMARY: A better understanding of the molecular pathways that regulate vascular smooth muscle cell function will provide further insights into the pathophysiological mechanisms that contribute to vascular changes and end-organ damage associated with high blood pressure, and could permit identification of potential novel therapeutic targets in the prevention and management of hypertension. PMID- 12589178 TI - Sympathetic control of the circulation in hypertension: lessons from autonomic disorders. AB - PURPOSE OF THE REVIEW: Inappropriate sympathetic overactivity is consistently observed in patients with essential hypertension. The present review summarizes the recent advances that have been made in our understanding of the role of the sympathetic nervous system in hypertension. RECENT FINDINGS: Studies in patients with autonomic disorders underscore the role of the sympathetic nervous system in the long-term maintenance of hypertension. Abnormalities in the afferent limb of the sympathetic nervous system, in the regulation of central neurons where sympathetic outflow originates, and in the modulation of efferent sympathetic function, can all produce autonomic disorders that are associated with hypertension. More subtle dysfunctions in any of these components have been described in essential hypertension and can contribute to its pathogenesis. These include impaired buffering capacity of arterial baroreflexes, increased central sympathetic outflow, and enhanced norepinephrine release (or decrease reuptake) from sympathetic nerve terminals. Whether genetic polymorphisms of adrenoreceptors are associated with essential hypertension is an area of active research. SUMMARY: Increased sympathetic activity can contribute to sustained hypertension not only because of its hemodynamic effects (increased cardiac output and vascular resistance), but also by altering renal and water handling by the kidney, and by inducing cardiac and vascular remodeling. Antihypertensive agents that directly or indirectly target this sympathetic overactivity may be particularly useful in long-term treatment of essential hypertension. PMID- 12589179 TI - Vascular inflammation: a role in vascular disease in hypertension? AB - PURPOSE OF REVIEW: In this review, we summarize the more recent clinical evidence highlighting the importance of vascular inflammation in terms of clinical risk prediction, and the mechanisms mediating the upregulation of inflammatory mediators in cardiovascular disease and hypertension. RECENT FINDINGS: Markers of inflammation have been shown to be upregulated in different forms of cardiovascular disease, and to correlate with vascular risk. Atherosclerosis is characterized by chronic inflammation of the vascular wall. The I-kappaB/nuclear factor-kappaB system is considered a major intracellular inflammatory pathway, mediating most of the vascular inflammatory responses. Increasing evidence indicates that hypertension, through the vasoactive peptides angiotensin and endothelin-1, promotes and accelerates the atherosclerotic process via inflammatory mechanisms. In animal and human studies proinflammatory properties of angiotensin II have been demonstrated in large conduit and small arteries, in the kidney as well as in the heart. The angiotensin II receptors involved in the inflammatory process and the interaction between angiotensin II and nitric oxide in mediating vascular inflammation have been identified. In addition, recent advances concerning the role of endothelin-1 as another important mediator of chronic inflammation in the vascular wall has been documented, and the relationship between endothelin-1 and angiotensin II on vascular inflammation demonstrated. SUMMARY: Inflammatory mechanisms are important participants in the pathophysiology of hypertension and cardiovascular disease. The identification of useful markers of inflammation, of new therapeutic targets to interfere with these mechanisms, and the evaluation of the efficacy of antiinflammatory treatments will allow progress in our ability to combat cardiovascular disease and the complications of hypertension. PMID- 12589180 TI - Dopamine and the kidney: a role in hypertension? AB - PURPOSE OF REVIEW: Defective transduction of the dopamine receptor signal in the kidney has been shown to be important in the pathogenesis of hypertension This review will discuss the genetic mechanism for the defective renal dopaminergic function and the interaction with other gene variant products in the pathogenesis of salt sensitivity and essential hypertension. RECENT FINDINGS: Single nucleotide polymorphisms of G protein-coupled receptor kinase type 4 (GRK4) phosphorylate, desensitize, and diminish the inhibitory action of D receptors on sodium transport in the kidney. Inhibition of GRK4 expression normalizes renal proximal tubule D receptor function in humans and rodents and ameliorates the hypertension in genetically hypertensive rats. Expression of the GRK4 variant, GRK4gammaA142V, produces hypertension and impairs the natriuretic effect of D receptor stimulation in mice. In humans, GRK4 single nucleotide polymorphisms are associated with essential hypertension, particularly salt sensitive hypertension. The prediction of the hypertensive phenotype is most accurate when elements of the renin-angiotensin system and GRK4 are included in the analysis. SUMMARY: GRK4 single nucleotide polymorphisms, by preventing the natriuretic function of the dopaminergic system and by allowing the antinatriuretic function of angiotensin II type 1 receptors to predominate, may be responsible for salt sensitivity. Hypertension develops with additional perturbations caused by the variants of other genes (e.g., alpha-adducin, angiotensin converting enzyme, angiotensinogen, angiotensin II type 1 receptor, aldosterone synthase, 11beta-hydroxysteroid dehydrogenase type 2), the quantitative interaction of which may vary depending upon the genetic background. PMID- 12589181 TI - Obesity-associated hypertension and kidney disease. AB - PURPOSE OF REVIEW: The worldwide prevalence of obesity and its associated metabolic and cardiovascular disorders has risen dramatically during the past two decades. Our objective is to review the mechanisms that link obesity with hypertension and altered kidney function. RECENT FINDINGS: Current evidence suggests that excess weight gain may be responsible for 65-75% of the risk for essential hypertension. Abnormal renal pressure natriuresis, due initially to increased renal tubular sodium reabsorption, is a key factor linking obesity with hypertension. Obesity increases renal sodium reabsorption by activating the renin angiotensin and sympathetic nervous systems, and by altering intrarenal physical forces. Adipose tissue functions as an endocrine organ, secreting hormones/cytokines (e.g. leptin) that may activate the sympathetic nervous system and alter kidney function. Excess visceral adipose tissue may physically compress the kidneys, increasing intrarenal pressures and tubular reabsorption. Sustained obesity eventually causes structural changes in the kidneys and loss of nephron function, further increasing arterial pressure and leading to severe renal disease in some cases. SUMMARY: Despite considerable progress in understanding the pathophysiology of obesity, there are still no specific guidelines for the treatment of obesity hypertension other than weight reduction. Special considerations for obese hypertensive patients, in addition to controlling blood pressure, are correcting the metabolic abnormalities and protecting the kidneys from injury. This remains an important area for further research, especially in view of the current 'epidemic' of obesity in most industrialized countries. PMID- 12589182 TI - Fat as a physiological regulator: the news gets better. PMID- 12589183 TI - Lipids and the immune response: from molecular mechanisms to clinical applications. AB - PURPOSE OF REVIEW: This review critically evaluates recent studies investigating the effects of fatty acids on immune and inflammatory responses in both healthy individuals and in patients with inflammatory diseases, with some reference to animal studies where relevant. It examines recent findings describing the cellular and molecular basis for the modulation of immune function by fatty acids. The newly emerging area of diet-genotype interactions will also be discussed, with specific reference to the anti-inflammatory effects of fish oil. RECENT FINDINGS: Fatty acids are participants in many intracellular signalling pathways. They act as ligands for nuclear receptors regulating a host of cell responses, they influence the stability of lipid rafts, and modulate eicosanoid metabolism in cells of the immune system. Recent findings suggest that some or all of these mechanisms may be involved in the modulation of immune function by fatty acids. SUMMARY: Human studies investigating the relationship between dietary fatty acids and some aspects of the immune response have been disappointingly inconsistent. This review presents the argument that most studies have not been adequately powered to take into account the influence of variation (genotypic or otherwise) on parameters of immune function. There is well documented evidence that fatty acids modulate T lymphocyte activation, and recent findings describe a range of potential cellular and molecular mechanisms. However, there are still many questions remaining, particularly with respect to the roles of nuclear receptors, for which fatty acids act as ligands, and the modulation of eicosanoid synthesis, for which fatty acids act as precursors. PMID- 12589184 TI - High- versus low-fat diets in human diseases. AB - PURPOSE OF REVIEW: To evaluate the evidence with regard to high-versus low-fat diets in the context of the prevention and management of obesity, type 2 diabetes and coronary heart disease. RECENT FINDINGS: Despite the increasing prevalence of obesity, there is no evidence to support the view that this is caused by an increased intake of fat. Fat sensors play an important role in regulating energy balance and lipid metabolism, and hypoenergetic diets containing 30-35% energy from fat promote weight loss. High intakes of carbohydrates with a high glycaemic index can result in insulin resistance, but this effect can be modulated by increased physical activity. SUMMARY: Although arguments to decrease the intake of trans and saturated fatty acids are cogent, the scientific basis for a reduction in the proportion of energy from fat below 30% energy is not supported by experimental evidence. A modest reduction in fat intake to 30-35% energy, with the bulk of carbohydrates being derived from complex carbohydrates from unrefined sources, would appear to be the best option for the prevention of obesity and cardiovascular disease. Increased physical activity appears to be particularly important in modulating the adverse effects associated with high-carbohydrate low fat diets. PMID- 12589185 TI - Oxidized fats in foods. AB - PURPOSE OF REVIEW: Lipid oxidation is the cause of important deteriorative changes in chemical, sensory and nutritional food properties. In particular, the question of whether oxidized fats in the diet may be detrimental to health is nowadays of the upmost concern, but finding an answer is not easy and requires careful consideration of different aspects of lipid oxidation. RECENT FINDINGS: In this review, the most recent works on the formation, nature and evaluation of oxidized dietary lipids are addressed; important issues such as the difficulties encountered in estimating their intake and the relationships between oxidants and antioxidants in the diet are discussed, and the latest studies on health implications of oxidized lipids are summarized. SUMMARY: The current literature reflects various important points. At present, there is no information on the intake of oxidized fats, which is essential to know if the amount of oxidized lipids in normal diets is sufficient to cause the physiological effects claimed. Recently, relevant advances in analytical methodologies for quantitation of specific oxidation compounds have been reported, although their application to improve the analytical definition of the oxidized substrate used in nutritional studies is still a goal to be reached. Alternatively, one of the most promising current tendencies in this field is the study of the molecular targets by which dietary oxidized lipids can influence health. Overall, more selected research based on coordinated multidisciplinary studies is needed to define the role of dietary oxidized fats in health. PMID- 12589186 TI - Fat versus carbohydrate in insulin resistance, obesity, diabetes and cardiovascular disease. AB - PURPOSE OF REVIEW: This review assesses the relative effect of fat versus carbohydrate and the differences between fatty acids and types of carbohydrate on insulin resistance and associated risk factors for diabetes and cardiovascular disease. RECENT FINDINGS: The debate continues over whether high-carbohydrate or high-fat diets have the more deleterious metabolic effects. Large randomized controlled trials have shown that a reduction of fat intake as part of a healthy lifestyle combined with weight reduction and exercise reduce the risk of type 2 diabetes. Carbohydrate as fruit and vegetable together with low-fat dairy products reduce blood pressure. The results of trials of fatty acid type continue to favor the use of monounsaturated fats. However, the advantages over carbohydrate have not always been clear. In terms of carbohydrate, the glycemic index appears to be a better predictor of the metabolic effects of a diet than the sugar content. The fiber content of the carbohydrate food appears to confer benefits in terms of diabetic control. Lower cholesterol and postprandial blood glucose results are associated with viscous fibers. SUMMARY: Diets that are higher in monounsaturated fatty acids, fiber and low glycemic index foods appear to have advantages in insulin resistance, glycemic control and blood lipids in a number of studies. The division of nutrients into total fat (regardless of fatty acids) versus carbohydrate (type and quantity not specified) appears to be less helpful in predicting outcomes. PMID- 12589187 TI - Apolipoprotein and apolipoprotein receptor genes, blood lipids and disease. AB - PURPOSE OF REVIEW: Apolipoproteins and their receptors are the main controllers of lipid metabolism and, as such, have a major impact not only on the risk of cardiovascular disease but also on the development and degeneration of the central nervous system. Variations in the genes coding for these apolipoproteins and their receptors and the interaction with the environment determine individual susceptibility to metabolic disturbances, the response to dietary or pharmacological intervention and, finally, to disease. RECENT FINDINGS: This review will focus on recent findings, such as the latest concepts regarding apolipoprotein E in neurodevelopment, the newly identified apolipoprotein A-V and its influence in triglyceride metabolism, and the improved understanding of apolipoprotein A-I and HDL metabolism in the light of the discovery of the ABC family of transporters. Other key aspects of lipoprotein metabolism and cardiovascular disease risk such as apolipoprotein B-100, the LDL receptor, apolipoprotein C-III or apolipoprotein (a) will be updated. SUMMARY: Variations in these genes will be analysed in relation to plasma lipid levels, their interactions with diet, treatment or other environmental stimuli, and their influence on the risk of cardiovascular disease and neurological disorders. PMID- 12589188 TI - Immune-enhancing diets for stressed patients with a special emphasis on arginine content: analysis of the analysis. AB - PURPOSE OF REVIEW: After two decades of intensive research, whether arginine enriched diets for oral/enteral administration are beneficial or harmful for stressed patients remains uncertain. An American consensus and a meta-analysis provide divergent conclusions. The main goal of the present review is to analyze these documents. RECENT FINDINGS: The so-called immune-enhancing diets have been found to be beneficial to postoperative patients. Nothing proves, however, that arginine is responsible for these beneficial effects since immune-enhancing diets contain other pharmacologically active components (e.g. omega3 free fatty acids, RNAs, antioxidant vitamins). In fact, arginine-enriched diets may be harmful in hemodynamically unstable patients and those presenting with multiple organ failure. SUMMARY: In light of the current doubts and until convincing data are produced, immune-enhancing diets should not be used in unstable critically ill patients. PMID- 12589189 TI - Metabolic and nutritional support in acute cardiac failure. AB - PURPOSE OF REVIEW: Cardiovascular disease is one of the most important causes of morbidity and mortality in western countries, generating an increasing number of admissions to intensive care units. Cardiac failure has long been associated with nutritional disorders, malnutrition and cachexia being frequent during the late phases of congestive heart failure: undernutrition is also a determinant of outcome, even after cardiac transplantation. RECENT FINDINGS: It has been shown that early metabolic support can improve the recovery of the ischaemic heart. This paper reviews recent findings on substrates that can support the failing myocardium, which are mainly glucose-insulin, glutamine, taurine, selenium, thiamine, folic acid, and omega-3 fatty acids. Ischaemia-reperfusion generates tissue lesions that can be partly prevented through substrate manipulation. SUMMARY: Shifting the substrate metabolism from lipids to carbohydrates and reinforcing the antioxidant status reduces the deleterious biological and clinical consequences of acute ischaemic events. The use of the glucose-insulin potassium infusion has become widespread with the re-discovery of its value in modulating cellular metabolism and accelerating recovery of the ischaemic myocardium. Antioxidants have gained acceptance in the perioperative phase, as well as in chronic heart failure. This constitutes another piece of evidence in favour of early metabolic and nutritional intervention. There also appears to be room for the prevention of acute deterioration of cardiac function after surgery with the preoperative administration of oral supplements containing omega-3 fatty acids. PMID- 12589190 TI - Nutrition support in critically ill septic patients. AB - PURPOSE OF REVIEW: Infection and sepsis remain major challenges in the critically ill. How nutritional therapy can effect real clinical outcomes is not easily apparent from the clinical data. Despite reducing infectious incidence, many studies show little difference in meaningful clinical outcomes. RECENT FINDINGS: Confusion over the role that nutrition and nutrition practice in intensive care plays in preventing infection, and later how nutrition aids recovery from sepsis perhaps explains many of the misconceptions and difficulties with the evidence. SUMMARY: Encouraging new evidence will help our decision making and shows that outcome can be improved by performing relatively simple therapies well. PMID- 12589191 TI - The role of visceral proteins in the nutritional assessment of intensive care unit patients. AB - PURPOSE OF REVIEW: This review analyses the recently published literature focusing on nutritional assessment in intensive care unit patients. The metabolic response to nutritional intervention is difficult to evaluate in critically ill patients whose body weight is influenced largely by massive fluid administration or losses. Visceral protein plasma levels have been proposed for this purpose, because they reflect hepatic synthesis in response to nutrient supply. However, in acute inflammatory states, liver activity is converted to the synthesis of acute-phase response proteins, resulting in a dramatic drop in visceral proteins, despite nutritional support. RECENT FINDINGS: The data regarding visceral protein levels were examined in relation to nutritional supplementation, and compared with other nutritional parameters and clinical outcomes. Transthyretin and retinol-binding protein levels seem to be the most sensitive to nutritional intervention. They are also the earliest to rise at the decrease of acute-phase protein levels, therefore representing a good index of the reversing reprioritization of hepatic protein synthesis. An inconsistent relationship was found between visceral protein plasma levels and clinical outcome in intensive care unit patients, probably because of the difficulty in demonstrating clearly a beneficial effect of nutritional supplementation in highly catabolic conditions. SUMMARY: In the acute stage of critical illness, the bi-weekly measurement of transthyretin together with acute-phase response protein plasma levels seems to be a 'window' on the metabolic condition (anabolism versus catabolism). However, only in the presence of stable inflammatory parameters do transthyretin levels reflect the adequacy of nutritional coverage. PMID- 12589192 TI - Role of L-glutamine in critical illness: new insights. AB - PURPOSE OF REVIEW: This review will attempt to summarize recent clinical and experimental data on glutamine's use in critical illness. It will try to present the concept of glutamine as a 'drug' or 'nutraceutical', given in addition to standard nutritional support. RECENT FINDINGS: Glutamine, traditionally considered to be a nonessential amino acid, is now considered as 'conditionally essential' following critical illness and injury. States of critical illness lead to significant decreases in plasma levels of glutamine and when this decrease is severe it has been correlated with increased mortality. Laboratory data have demonstrated numerous benefits of glutamine in experimental models of critical illness, including attenuated proinflammatory cytokine expression, improved gut barrier function, enhanced ability to mount a stress response, improved immune cell function, and decreased mortality. Over the last 10 years clinical trials of glutamine supplementation in critical illness have shown benefit with regard to mortality, length of stay, and infectious morbidity. However, data demonstrating a lack of benefit with glutamine supplementation in the critically ill have been presented as well. It appears that dose and route of administration clearly influence the benefit observed from glutamine administration, with high-dose, parenteral glutamine demonstrating an advantage over low-dose, enteral glutamine. SUMMARY: High-dose or parenteral (>0.20-0.30 g/kg/day or > or =30 g/day) glutamine appears to demonstrate the greatest potential for benefit in critically ill patients. No evidence of harm has been observed in studies conducted to date, thus further clinical trials utilizing glutamine as a pharmacologic supplement to standard nutrition are warranted. PMID- 12589193 TI - Arginine availability, arginase, and the immune response. AB - PURPOSE OF REVIEW: Arginine, often found in immunonutrition regimens, is an important modulator of immune system activation. However, the mechanism of how arginine may be beneficial in immunonutrition is poorly understood. This review details the importance of arginine, its metabolism, and ultimately, its physiologic role in critically ill and immunocompromised patients. RECENT FINDINGS: The metabolism of arginine is determined by the expression of the arginine metabolizing enzymes inducible nitric oxide synthase and two arginase isoforms (arginase I and II). Inducible nitric oxide synthase is induced by T helper I cytokines (interleukin-1, tumor necrosis factor and gamma-interferon), while arginases are induced by T helper II cytokines and other immune regulators such as interleukins 4, 10, and 13, transforming growth factor-beta and prostaglandin E2. Endotoxin induces inducible nitric oxide synthase and arginases I and II. Arginase plays an important role in the production of ornithine, a precursor of proline and polyamines, both of which are necessary for cellular proliferation and wound healing. Arginase also induces nitric oxide synthase activity by competing for arginine availability in the extracellular environment, and producing polyamines, which may modulate macrophage activation. Through limitation of arginine availability in the extracellular environment, arginases also potentially regulate other 'arginine-dependent' immune functions such as T lymphocyte activation, although this hypothesis remains to be proven. SUMMARY: The availability of arginine during critical illness may be regulated by arginase activity. Thus, arginase expression appears to be essential in the regulation of the cellular immune response and the inflammatory process during critical illness. PMID- 12589194 TI - L-Glycine: a novel antiinflammatory, immunomodulatory, and cytoprotective agent. AB - PURPOSE OF REVIEW: In recent years, evidence has mounted in favor of the antiinflammatory, immunomodulatory and cytoprotective effects of the simplest amino acid L-glycine. This article will focus on the recent findings about the responsible mechanisms of protection and review the beneficial effects of glycine in different disease states. RECENT FINDINGS: Glycine protects against shock caused by hemorrhage, endotoxin and sepsis, prevents ischemia/reperfusion and cold storage/reperfusion injury to a variety of tissues and organs including liver, kidney, heart, intestine and skeletal muscle, and diminishes liver and renal injury caused by hepatic and renal toxicants and drugs. Glycine also protects against peptidoglycan polysaccharide-induced arthritis and inhibits gastric secretion and protects the gastric mucosa against chemically and stress induced ulcers. Glycine appears to exert several protective effects, including antiinflammatory, immunomodulatory and direct cytoprotective actions. Glycine acts on inflammatory cells such as macrophages to suppress activation of transcription factors and the formation of free radicals and inflammatory cytokines. In the plasma membrane, glycine appears to activate a chloride channel that stabilizes or hyperpolarizes the plasma membrane potential. As a consequence, agonist-induced opening of L-type voltage-dependent calcium channels and the resulting increases in intracellular calcium ions are suppressed, which may account for the immunomodulatory and antiinflammatory effects of glycine. Lastly, glycine blocks the opening of relatively non-specific pores in the plasma membrane that occurs as the penultimate event leading to necrotic cell death. SUMMARY: Multiple protective effects make glycine a promising treatment strategy for inflammatory diseases. PMID- 12589228 TI - Hemochromatosis and Iron Overload Screening (HEIRS) study design for an evaluation of 100,000 primary care-based adults. AB - BACKGROUND: The HEIRS Study will evaluate the prevalence, genetic and environmental determinants, and potential clinical, personal, and societal impact of hemochromatosis and iron overload in a multiethnic, primary care-based sample of 100,000 adults over a 5-year period. Participants are recruited from 5 Field Centers. Laboratory testing and data management and analysis are performed in a Central Laboratory and Coordinating Center, respectively. METHODS: Participants undergo testing for serum iron measures and common mutations of the hemochromatosis gene ( ) on chromosome 6p and answer questions on demographics, health, and genetic testing attitudes. Participants with elevated values of transferrin saturation and serum ferritin and/or C282Y homozygosity are invited to undergo a comprehensive clinical examination (CCE), as are frequency-matched control subjects. These examinations provide data on personal and family medical history, lifestyle characteristics, physical examination, genetic counseling, and assessment of ethical, legal, and social implications. Primary and secondary causes of iron overload will be distinguished by clinical criteria. Iron overload will be confirmed by quantification of iron stores. Recruiting family members of cases will permit DNA analysis for additional genetic factors that affect iron overload. RESULTS: Of the first 50,520 screened, 51% are white, 24% are African American, 11% are Asian, 11% are Hispanic, and 3% are of other, mixed, or unidentified race; 63% are female and 37% are male. CONCLUSIONS: Information from the HEIRS Study will inform policy regarding the feasibility, optimal approach, and potential individual and public health benefits and risks of primary care based screening for iron overload and hemochromatosis. PMID- 12589229 TI - Orthostatic intolerance is not necessarily related to a specific mutation (Ala457Pro) in the human norepinephrine transporter gene. AB - BACKGROUND: Orthostatic intolerance (OI) is a syndrome characterized by lightheadedness, palpitations, fatigue, blurred vision, dizziness, chest discomfort, cognitive impairment, and occasionally syncope. These symptoms usually occur after upright posture and are associated with tachycardia and high plasma concentrations of norepinephrine. It has been proposed that a mutation in exon 9 of the norepinephrine transporter gene (Ala457Pro), resulting in more than 98% loss of function compared with the wild type, might provide a pathogenetic mechanism to explain the clinical symptoms of patients with OI. METHODS: We studied 46 young men from military service who had sought medical advice because of dizziness while standing. Every patient underwent a tilt-table test, with monitoring of blood pressure, heart rate, and plasma catecholamines in supine position and during 30 minutes of standing. Fourteen patients showing the full blown OI syndrome (30 bpm increase in heart rate and 600 pg/mL plasma norepinephrine levels while standing) underwent direct DNA sequencing of exon 9 of the norepinephrine-transporter gene. RESULTS AND CONCLUSIONS: The specific mutation (Ala457Pro) was not detected in any of the 14 OI patients. Based on these findings, we doubt that this specific genetic transport defect is a frequent cause of the impaired uptake of norepinephrine in OI patients. Its routine determination will therefore not be helpful to establish the clinical diagnosis of OI. PMID- 12589230 TI - Beneficial effects of a glyburide/metformin combination preparation in type 2 diabetes mellitus. AB - BACKGROUND: Type 2 diabetes mellitus is characterized by both insulin deficiency and insulin resistance. Effective treatment often requires therapy directed at both abnormalities. Patients on monotherapy might benefit from a combination agent such as glyburide/metformin, which increases insulin secretion and reduces insulin resistance. METHODS: All patients taking a glyburide/metformin preparation at the Carl T. Hayden VAMC were identified from pharmacy records. Patients with documented hemoglobin A values within 31 weeks prior and between 3 and 33 weeks after initiation of therapy (92 subjects) were examined. RESULTS: Glyburide/metformin combination therapy reduced hemoglobin A levels from 0.087 to 0.083 (P < 0.06). Significant reductions were seen in those patients with initial levels higher than 0.08 (0.094 to 0.087; P < 0.01). No significant reductions were seen in those patients with initial levels lower than 0.08. CONCLUSIONS: In patients on monotherapy or on dual oral therapy with inadequate control, changing to a glyburide/metformin combination preparation may improve glucose control. PMID- 12589231 TI - A 50-year-old man with chest pain. PMID- 12589232 TI - New indications for treatment of chronic inflammation by TNF-alpha blockade. AB - The impressive anti-inflammatory effects of the tumor necrosis factor (TNF)alpha blockers etanercept and infliximab have led to their use in multiple inflammatory diseases besides their original indication, rheumatoid arthritis (RA). The well studied clinical effects of both agents in RA are the reduction of signs and symptoms of joint inflammation as well as the arrest of bone destruction. Infliximab has also been Food and Drug Administration-approved in the treatment of Crohn disease; etanercept is now FDA-approved for juvenile chronic arthritis and psoriatic arthritis. Favorable initial clinical trials have been reported in other rheumatic diseases, including ankylosing spondylitis and adult Still disease. In addition, TNF alpha blockade is being studied in the treatment of uveitis, myelodysplastic syndromes, and graft-versus-host disease. Studies in sepsis and septic shock have identified small subsets of patients that may benefit from TNF alpha blockade, but broader use in septic patients has not improved survival. The TNF alpha blockers have had relatively infrequent serious side effects, especially compared with the immunosuppressive and cytotoxic agents otherwise employed to treat these diseases. Further studies of optimal dosing, combination with other therapies, and long-term benefits and side effects will emerge from future trials. PMID- 12589233 TI - A patient with sodium- and potassium-losing nephropathy. AB - A young man with salt-losing nephropathy had the unusual coexistence of hypokalemia caused by secondary hyperaldosteronism. When his NaCl intake was supplemented by 12 g/day (205 mmol), hyperaldosteronism was suppressed and so were his urinary K wasting, hypokalemia, and episodic falling, during an extended follow-up. His findings are compared and contrasted with those in other reported patients with renal salt wasting, and also those in patients with Gitelman and Bartter syndromes. PMID- 12589234 TI - Monoclonal gammopathy and spurious hypophosphatemia. AB - BACKGROUND: Spuriously low levels of plasma phosphate have been reported previously in patients with multiple myeloma and polyclonal gammopathy. We report 2 cases of spurious hypophosphatemia in patients with elevated concentrations of serum monoclonal immunoglobulins, 1 of whom had monoclonal gammopathy of undetermined significance and the other multiple myeloma. METHODS: Plasma phosphate concentrations were measured using nondeproteinized and deproteinized plasma samples from patients with monoclonal gammopathies. RESULTS: In 2 patients with monoclonal gammopathy, the levels of plasma inorganic phosphate were reported as <1.0 mg/dL when the phosphate concentration was determined using an analyzer that employs nondeproteinized plasma. When the samples were reanalyzed using a laboratory method that removes serum proteins, normal or elevated concentrations of phosphate were found. Plasma levels of phosphate in 4 other patients with monoclonal gammopathy were normal by both methods. CONCLUSIONS: These data confirm previous reports that spurious hypophosphatemia occurs in some patients with increased levels of serum monoclonal immunoglobulins when laboratory methods using nondeproteinized samples are employed. The occurrence of unusually low plasma phosphate concentrations in patients without symptoms or clinically apparent causes of hypophosphatemia should alert physicians to search for monoclonal gammopathy. PMID- 12589235 TI - Churg-Strauss syndrome: diagnostic difficulties and pathogenesis. AB - A 52-year-old woman with a history of chronic obstructive pulmonary disease presented with symmetrical polyarthritis involving her metacarpophalangeal and proximal interphalangeal joints, knees, ankles, and hips and with a purpuric rash involving her lower extremities. She had a history of recurrent episodes of purulent otitis often associated with myalgias and arthralgias. Laboratory studies at presentation included leukocytosis with 16% eosinophils, an elevated rheumatoid factor titer, and an elevated antineutrophil cytoplasmic antibody titer. Cultures from the right ear canal grew. Skin biopsy revealed leukocytoclastic vasculitis with pericapillary eosinophils. The patient was treated with prednisone and then with azathioprine after the rash relapsed during the tapering of prednisone. Four months after her initial presentation, she developed bilateral foot drop. A sural nerve biopsy revealed vasa nervosum vasculitis. The diagnosis of Churg-Strauss syndrome was established, and she was treated with an increased dose of azathioprine and a slowly tapering prednisone regimen. This case report suggests that patients with Churg-Strauss syndrome can present with a syndrome suggesting rheumatoid arthritis. In this particular patient, recurrent staphylococcal infections may have triggered the vasculitic process. PMID- 12589236 TI - Evidence-based nursing and community practice. PMID- 12589237 TI - Smoking cessation: working in partnership. PMID- 12589238 TI - A protocol for treating acute constipation in the community setting. AB - Constipation is a distressing symptom, common in many patient groups and traditionally treated by nurses. Ambiguity concerning limits of the nurse's role in treating constipation exists and guidelines are scarce. Theoretically constipation is largely preventable, but acute episodes arise for a number of reasons and often need prompt treatment. This article describes the development of a local protocol which empowers appropriately trained community nurses to treat adult patients with acute constipation. A flow chart was used to define the guidelines. Implementation of the protocol is briefly discussed in relation to other current local initiatives. PMID- 12589239 TI - Legal issues arising in community nursing 5: elder abuse. PMID- 12589240 TI - An overview of the development of nurse prescribing. AB - This article provides an overview of nurse prescribing in the UK. The initial recommendations for nurse prescribing, perceived benefits of prescribing and the findings from evaluation studies are reported. Current education and training courses available for nurse prescribers are described. The development of nurse prescribing has been slow and, it could be argued, out-dated, as many practitioners are excluded from this initiative. It is important that nurse prescribers receive adequate time and funding to undertake appropriate training and education in this area. PMID- 12589241 TI - Pharmacist's perceptions of nurse prescribing of emergency contraception. AB - Proposed strategies to reduce the rate of unplanned pregnancy in the UK include widening access to hormonal emergency contraception (HEC) through nurse prescribing and over-the-counter sales by pharmacists. This article reports on the perceptions of pharmacists regarding the appropriateness of nurse prescribing of HEC drawn from a survey of pharmacists' attitudes towards HEC deregulation. A 10% sample (n = 3999) of all members of the Royal Pharmaceutical Society of Great Britain was surveyed and a response rate of 38.6% (1543 respondents) achieved. Pharmacists were asked whether or not nurses should prescribe HEC and in what capacity. In respect of the statement that: 'Nurses should not be allowed to prescribe post-coital contraception', 42% of respondents agreed while 31.6% disagreed. This indicates that pharmacists do not have a consensus view regarding nurse prescribing of HEC. Arrangements for pharmacists prescribing under protocol received the support of 60.2% of respondents, whereas 41% expressed support for nurse prescribing under protocol. There was also relatively little support for 'specifically trained' nurses prescribing HEC. The findings are considered in relation to the development of a coherent and accessible provision of HEC and the wider implications for interprofessional relationships concerning prescribing. PMID- 12589242 TI - A review of the psychosocial aspect of multiple sclerosis. AB - The psychosocial aspects of multiple sclerosis are far-reaching and may have devastating effects on individuals and carers alike. Issues such as the impact of uncertainty, the need to maintain hope and control, and a person's ability to adjust his/her lifestyle to the disease progression are highlighted by recent research. The individualized nature of this chronic illness experience makes comprehensive and ongoing assessment crucial. Evidence from practice suggests that the district nurse's contribution to psychosocial support is hindered by poor role definition. Similarly, while the literature acknowledges the benefits of empowerment, difficulties exist in practice. Collaboration between district nurses and clinical nurse specialists is advocated as a means of integrating best practice and clinical knowledge with community expertise. The need for improved research methodology is identified. PMID- 12589243 TI - A study of health services for the Chinese minority in Manchester. AB - This article describes a small-scale study which examined the views of both users and providers of primary healthcare services for the Chinese minority in Manchester. Thirty Chinese mothers of pre-school children were selected from immunization records and interviewed face-to-face at their homes using structured questionnaires and unstructured discussions. Thirty-eight GPs who had the largest number of Chinese patients (n = 10) registered with them, and 26 health visitors who worked with these GPs were also interviewed, using semi-structured questionnaires. The findings indicate that a discrepancy exists between the views of GPs, health visitors and Chinese mothers on the use of primary healthcare services. The health professionals' awareness of the effects of socio-economic characteristics, demographic profile and indigenous health beliefs and attitudes on the quality of primary healthcare services for Chinese families in Manchester is discussed. PMID- 12589244 TI - Primary care research needs community nurses. PMID- 12589245 TI - Pharmacy proposals show muddled thinking. PMID- 12589246 TI - Barriers to smoking cessation in pregnancy: a qualitative study. AB - The magnitude of smoking as a public health concern in Britain is reflected in the recent government investment to develop smoking cessation services. Smoking in pregnancy is a priority. This article presents the findings of a small qualitative research study exploring barriers to pregnant women stopping smoking. The research was conducted in deprived areas of South Yorkshire, with levels of smoking-related ill health higher than the national average. The study participants revealed a belief system resulting in a self-fulfilling prophecy of relapse or failure to quit. The role and meaning of smoking for women with a high caring burden and socio-economic problems resonate strongly with earlier studies. Other barriers were the influence of family and friends, how women interpreted facts relating to smoking risks and the nature of smoking cessation service delivery. Community nursing staff have a potential in the delivery of smoking cessation interventions. Effective training should be a priority. PMID- 12589247 TI - Accountability and drug administration in community care. AB - Medicines are a key therapeutic intervention in community care but their adverse effects can give rise to concern for patient safety. This article considers the legal regulation of medicines with emphasis on the right to administer medicines and the standard of administration. It further considers the consequences for primary care staff of the inappropriate use of widespread practices such as tablet crushing and covert administration of medicine. PMID- 12589248 TI - District nurses' experiences and perceptions of cancer patient referrals. AB - Traditionally palliative care of cancer patients has been seen as an important and defining aspect of district nursing. Care of the dying patient has been used to describe the ability and scope of the district nursing service to holistically provide for patient need. However health and social service changes in the last decade have refocused the district nurse's role away from holistic delivery to more specialized care. There is also research evidence that there are several contradictions in referral processes to the district nursing service. We conducted a study to explore district nurses' perceptions and experiences of referral of cancer patients to gain insight into these referral processes. We interviewed 20 nurses from three primary care trusts to explore the referral process of cancer patients to the district nursing service from the district nurses' point of view. The nurses expressed concerns regarding completeness, accuracy and appropriateness of referral and suggested improvements that could be made. PMID- 12589249 TI - How effective is training in compression bandaging techniques? AB - The aim of this review was to investigate whether formalized compression bandage training is required for nurses to achieve compression bandaging of high quality, that is bandaging which produces sustained graduation from toe to knee. Medline, CINAHL and Embase databases were used in the search. Randomized controlled trials and quasi-experimental trials were included where (at a minimum) nurses experienced in compression bandaging had their skills reviewed before and after training, and then reviewed once more at a later follow-up session. Three studies matched these criteria. In all three, nurses in general did not achieve sustained graduated compression before training, despite being defined as experienced bandagers. There was a substantial improvement in bandaging skills immediately following training. Two trials found that this improvement was sustained for between 2 and 4 weeks. The results of the third study suggested that improvements might not be sustained in the longer term. This review raises issues for further research including the current standard of compression bandaging, by those individuals who consider themselves competent in this skill. In addition, there needs to be argument on how nurses' bandaging skills can be improved and maintained, including the depth of role of feedback. In the meantime, nurses need to be as diligent as possible in maintenance of their compression bandaging skills, and the training they give junior staff. The effectiveness of current training cannot be assessed by short-term improvements and must not be assumed. PMID- 12589250 TI - Older people as health service consumers 2: health and illness in a village context. AB - Involving users in the future shaping and development of health care services is part of the drive to restore public confidence in the NHS (NHS Executive 2002). A fundamental part of this process is working in partnership with patients, and attempting to include marginalized groups. Illness, infirmity and lack of knowledge may prevent older people from being actively involved in decisions about health care services, and the culture to which they belong may act as a further barrier. For true partnership and patient involvement, there needs to be a common understanding of the meaning of health and illness. This article discusses the prevailing cultural health beliefs within an elderly, white village community in South England. PMID- 12589251 TI - Learning ought to follow mistakes. PMID- 12589253 TI - Peripheral benzodiazepine receptor: structure and function in health and disease. AB - In vitro studies using biochemical, pharmacological and molecular approaches demonstrated that the peripheral-type benzodiazepine receptor (PBR) is a mitochondrial protein, involved in the regulation of cholesterol transport from the outer to the inner mitochondrial membrane, the rate-determining step in steroid biosynthesis. In vivo animal models and ontogeny studies validated the role of PBR in steroidogenesis. Targeted disruption of the PBR gene in Leydig cells resulted in the arrest of cholesterol transport into mitochondria and steroid formation. Molecular modeling of PBR suggested that it might function as a channel for cholesterol. Indeed, cholesterol uptake and transport by bacteria cells was induced upon PBR expression. Amino acid deletion and site-directed mutagenesis studies identified a cholesterol recognition/interaction amino acid consensus sequence in the cytoplasmic carboxy-terminus of the receptor. In vitro reconstitution experiments demonstrated that the 18 kDa PBR protein binds with high affinity both drug ligands and cholesterol, suggesting that this protein might serve numerous functions considering the critical role of cholesterol in membrane biogenesis and human pathology. In this context, PBR expression correlated with the quality of kidney preservation, indicating that it might serve as an index of kidney and mitochondrial viability during ischemia reperfusion injury. PBR overexpression was also found to be a prognostic indicator of the aggressive phenotype in breast, colorectal and prostate cancers. Moreover, in Alzheimer's disease brain specimens, PBR levels were increased and paralleled the elevated neurosteroid synthesis observed in specific brain areas. The role for PBR in these pathological conditions remains to be elucidated. paralleled the elevated neurosteroid synthesis observed in specific brain areas. The role for PBR in these pathological conditions remains to be elucidated. PMID- 12589254 TI - [Synthesis and binding affinity of 3-aryl-7-hydroxycoumarins to human alpha and beta estrogen receptors]. AB - The synthesis of a set of substituted 3-aryl-7-hydroxycoumarins was performed. The study of the relations between their structure and their relative binding affinity (RBA) to human alpha and B estrogen receptors was achieved. PMID- 12589255 TI - [Synthesis and insecticidal activity of some benzimidazolic and benzothiazolic derivatives]. AB - The benzimidazolic and benzothiazolic derivatives with a (CH(2))n radical in 2 position of the heterocycle had been synthesized and then tested for their insecticidal activity on Culex pipiens larvae. The benzothiazolic derivatives were found to be the most active. The study has also showed that this activity was related to the nature of the substitute in 2 position of the heterocycle. This activity increased when (CH(2))n chain increased until n=6; above this value, the activity decreased. PMID- 12589256 TI - [In vitro uranyle affinity of per (3,6-anhydro-2-O-carboxymethyle)-alpha cyclodextrin and conditions required for in vivo application]. AB - Per (3.6-anhydro-2-O-carboxymethyle)- alpha-cyclodextrin ([1]) is a polydentate analog of EDTA, a well-known cation chelating reagent. [1] exhibits strong affinities in vitro for lanthanids, cobalt and also for uranyl cations. Hence, a 1:1 stoechiometry and a high affinity for uranyle (60.01; angular values are more physiological in the second position with pedal unit behind the saddle axis. CONCLUSIONS: The incidence and importance of low back pain in cyclists can be reduced with appropriate pedal unit position; the position with pedals behind the saddle axis permits more physiological spine angles in comparison with the classic one having the pedals in front of the saddle axis; this fact is due to a different pelvic position which coincides with lumbar angles. PMID- 12589271 TI - MRI demonstration of spinal cord ischemia in traumatic spinal injury without radiographic abnormalities. Case report. PMID- 12589272 TI - Ultrasound pattern of glomus tumor of the shoulder. PMID- 12589273 TI - Chronic granulomatosis mastitis. A case report. PMID- 12589274 TI - Granular cell tumor of the breast. Report of a case. PMID- 12589275 TI - Ingested medications reteined in the colon. PMID- 12589276 TI - A tamponade with a happy ending. PMID- 12589277 TI - An interdisciplinary approach for improved esthetic results in the anterior maxilla. AB - This clinical report describes an interdisciplinary (orthodontic, periodontic, and prosthodontic) approach for the coordinated treatment of a patient diagnosed with peg-shaped maxillary lateral incisors, diastemata, short clinical crowns, excessive gingival display, and orthodontic malocclusion. The patient's specific esthetic expectations for the anterior maxilla were successfully met through phased treatment, including orthodontic tooth movement, provisional restorations, periodontal surgery for lengthening of the clinical crown, porcelain laminate veneers, and direct composite restorations. Such coordinated interdisciplinary evaluation and treatment are necessary for improved esthetic results in maxillary anterior areas esthetically compromised in several aspects. PMID- 12589278 TI - Implant retained overdentures for two patients with severe lichen planus: a clinical report. AB - Lichen planus is a common inflammatory mucocutaneous disease that often manifests itself intraorally. Oral lichen planus can appear in many forms; the most significant form for the edentulous patient is the erosive variety. For the patient, wearing a complete denture is quite dramatic because of the friability of the tissue. Implant-supported overdentures are a predictable treatment for edentulous patients, but this has been discouraged for the patient with erosive lichen planus. This article describes 2 patients with oral erosive lichen planus who were successfully treated with implant-retained mandibular overdentures. PMID- 12589279 TI - The use of distraction osteogenesis to treat hemifacial microsomia: a clinical report. AB - Distraction osteogenesis is a method of forming bone through osteotomy and sequential stretching of the healing callus. This process is accomplished with the aid of a distraction device, which is secured with screws placed directly into bone for a predetermined length of time. This clinical report describes the use of distraction osteogenesis to treat mandibular asymmetry in a patient with Goldenhar's syndrome, a form of hemifacial microsomia. After treatment, the patient had a more rounded facial contour and corrected occlusal plane. PMID- 12589280 TI - Use of computed tomography for the fabrication of a custom brachytherapy carrier: a clinical report. AB - Early methods of fabricating customized radiation carriers for inaccessible areas, such as the nasopharyngeal space, normally required the patient to be under conscious sedation or general anesthesia to allow impressions for indirect processing techniques. This article describes the use of computed tomography to design and fabricate a carrier for the intracavity application of iodine 125 seeds for a patient with rhabdomyosarcoma of the nasopharynx. With tomography printed copies, a wax pattern was formed, flasked, boiled out, and eventually replaced with silicone material. The patient had to be available only for a conventional thin-cut computed tomography scan without contrast; no fitting appointments or anesthesia were necessary. The indirect procedure eliminated discomfort from impression making and allowed for precise fabrication of the nasopharyngeal carrier. The described technique can be used for other intracavitary radiation applications. PMID- 12589281 TI - A comparison of the depths produced using three different tooth preparation techniques. AB - STATEMENT OF PROBLEM: Inadequate preparation of teeth for metal ceramic crowns can significantly influence the ultimate form and longevity of the definitive restoration, as well as adversely affect the supporting tissues. PURPOSE: The purpose of the study was to determine the effect that 3 different tooth preparation techniques had on an operator's ability to appropriately and consistently prepare teeth for metal ceramic crowns. MATERIAL AND METHODS: Thirty typodont maxillary central incisor teeth were mounted individually and randomly allocated to 3 equal groups (A, B, and C). One operator was asked to prepare each tooth for a metal ceramic crown. A freehand approach was used to prepare the teeth in Group A, which acted as a control. Groups B and C were prepared with the assistance of a silicone index and a suitable depth gauge bur, respectively. A silicone index of the unprepared tooth, into which contrasting silicone was injected to occupy the space created by tooth preparation, was sectioned in the midline. Images of the sectioned index were captured with an optical microscope (resolution +/- 0.02 mm), which was attached to a personal computer. A calibrated image analysis program was used to measure the depth of preparation (in millimeters) at 5 points (labial cervical, mid-labial, incisal, mid-palatal, and palatal cervical) on 2 occasions. These results were pooled and averaged to give a mean labial and palatal preparation depth (in millimeters) and incisal edge reduction. The data were analyzed by use of a 1-way analysis of variance and Scheffe's post hoc statistical test (P<.05). RESULTS: The mean depths of labial and palatal preparation for Groups A, B, and C were 1.28 and 0.47 mm; 1.46 and 0.56 mm; and 1.45 and 0.63 mm, respectively. The difference between the groups' labial preparation depth was not significant (P=.06), but the difference for palatal preparation depth was significant (P=.01). The mean incisal reduction was 3.00 mm for Group A, 2.74 mm for Group B, and 2.13 mm for Group C (P=.00). CONCLUSION: Within the limitations of this study, it was concluded that preparation of teeth for metal ceramic crowns without the use of devices to help gauge reduction depth can result in insufficient labial reduction and incisal overreduction. PMID- 12589282 TI - The influence of working cast residual moisture and temperature on the fit of vacuum-forming athletic mouth guards. AB - STATEMENT OF PROBLEM: The comfort and effectiveness of athletic mouth guards are believed to depend on their degree of fit to oral tissues. Vacuum-forming machines are simpler and less expensive than pressure-forming machines. However, it is thought that vacuum-formed mouth guards often do not exhibit adequate adaptation. PURPOSE: The purpose of this study was to evaluate the effects of cast residual moisture and temperature on the fit of athletic mouth guards made with a vacuum-forming machine. MATERIAL AND METHODS: A metal master model simulating the cross section of the maxillary molar region was used to form 20 working plaster casts. The casts represented 4 (5 specimens each) conditions before the forming of the mouth guard specimens: storing in a wet environment at room temperature and storing in a dry environment at room temperature, 5 degrees C, and 40 degrees C. Mouth guard specimens were fabricated with ethylene vinyl acetate sheets (3.8-mm thick) with the use of a vacuum-forming machine. Test casts were created by pouring hand-mixed type III dental stone into each of the mouth guard specimens. The differences in the sagittal cross-sectional heights at the line angle area of the test casts and the working casts were compared. This was achieved by superimposing their 3-dimensional images scanned by a laser scanner. The air permeability was also measured for the hand-mixed stone casts under wet and dry conditions, the vacuum-mixed stone cast, and the high-strength stone specimen. This was achieved by measuring the volume of transmitted air passing through the specimen in the testing tube. One-way analysis of variance with the Scheffe post hoc test (P <.05) was applied to determine the conditions of the working cast required to achieve the best fit. RESULTS: With regard to the fit of the mouth guard specimens to the working cast, those with dry and heated working casts showed a significantly better fit than those with wet working casts (P<.05). A significantly larger volume of transmitted air was found in the dry stone specimen (P<.05) followed by the dry high-strength stone cast and then the wet stone cast. CONCLUSION: Within the limitations of this study, residual moisture in the working cast was the most critical factor in determining the fit of the mouth guard made by vacuum-forming machines. The best fit was achieved when the working cast was thoroughly dried and its surface temperature was elevated. PMID- 12589283 TI - In vitro evaluation of fluoride varnish on overdenture abutments. AB - STATEMENT OF PROBLEM: No clinical study has evaluated the efficacy of fluoride varnish as a topical fluoride agent for the protection of overdenture abutments from dental caries. PURPOSE: This study assessed the efficacy of a fluoride varnish in protecting overdenture abutments from demineralization in an in vitro environment. MATERIAL AND METHODS: Eighty caries-free, extracted teeth were sectioned and domed immediately below the cementoenamel junction, simulating overdenture abutment preparations. Acid resistant nail varnish was painted on all surfaces except for a window (1 x 4 mm) on the prepared occlusal dentin surfaces. Teeth were randomly divided into 4 groups of 20 specimens: control, washed with deionized/distilled water; Daily Gel, treated with Karigel-N (5,000 ppm) for 4 minutes daily; Weekly Gel, treated with Karigel-N for 4 minutes weekly; and Weekly Varnish, treated with Duraflor (22,600 ppm) weekly. The varnish was removed using a blade 24 hours after each application to simulate the clinical loss of the varnish. Teeth were then placed in a cycle of demineralization (2.2 mMol/L CaCl(2)2H(2)O, 2.2 mMol/L KH(2)PO(4), and 50 mMol/L acetic acid at pH 4.3) for 6 hours and remineralization (1.5 mMol/L CaCl(2)2H(2)O, 0.9 mMol/L KH(2)PO(4), and 150 mMol/L KCl at pH 7.0 for 17 hours) for 21 days. Half the teeth in each group were brushed with no dentifrice for 10 seconds, twice daily. Teeth were sectioned at 100 +/- 10 microm buccolingually and evaluated under polarized light microscopy and contact microradiography. The depth of each lesion and the width of the remineralization bands were measured (mm). Analysis of variance models and T-tests were used to assess the effects of different treatments (alpha=.05). Duncan multiple range tests were then chosen as post hoc tests to evaluate the statistical significance of all pairwise comparisons. RESULTS: The control group had the deepest lesions (mean depth 400 +/- 39 microm). The lesions from the varnish and daily gel groups were significantly shallower than the lesions in both the control group and the weekly gel group. The remineralization bands in the varnish group were 52% wider than the control group bands. However, the daily gel group had the largest increase in band width (117% increase) compared with the control group. There were significant differences between all groups for net depth (lesion depth minus the remineralization band) (P<.0001). Daily application of fluoride gel was significantly more effective than the other treatments tested. Brushing was not a significant factor. CONCLUSION: Within the limitations of this in vitro study, treatment of overdenture abutment with various topical fluorides significantly inhibited demineralization and enhanced remineralization of cut occlusal dentin surfaces. Daily use of Karigel-N was the most effective treatment, followed by the weekly application of Duraflor varnish. PMID- 12589284 TI - Dimensional change in complete dentures fabricated by injection molding and microwave processing. AB - STATEMENT OF PROBLEM: Acrylic resin complete dentures undergo dimensional changes during polymerization. Techniques with injection molding and polymerization and microwave polymerization are reported to reduce these changes and thereby improve clinical fit. These dimensional changes need to be quantified. PURPOSE: The purpose of this study was to compare differences in dimensional changes of simulated maxillary complete dentures during polymerization and storage in water after injection molding and conventional polymerization, or microwave polymerization against a control of conventionally packed and polymerized simulated maxillary complete dentures. MATERIAL AND METHODS: Forty identical maxillary denture bases were prepared in dental wax with anatomic teeth. They were invested and the wax eliminated from the molds. Ten specimens each were randomly assigned to 1 of 4 groups. Group 1 was compression molded and conventionally polymerized; group 2 was injection molded and conventionally polymerized (Success); group 3 was injection molded and microwave polymerized (Acron MC); and group 4 was injection molded and microwave polymerized (Microbase). Intermolar width and changes in vertical dimension of occlusion, were determined after polymerization and after storage in water for 28 days. Measurements in triplicate were made between points scribed on the second molar teeth with a traveling microscope (accurate to 0.005 mm). Vertical dimension of occlusion was measured between points scribed on the upper and lower members of an articulator by use of an internal micrometer (accurate to 0.05 mm). Data were analyzed by use of a 1-way analysis of variance with Tukey post-hoc contrasts (P <.05). RESULTS: Polymerization contractions (intermolar widths) for each group were: group 1, -0.24%; group 2, -0.27%; group 3, -0.35%; and group 4, -0.37%. The Microbase specimens had greater shrinkage than conventionally polymerized specimens, but there were no significant differences between the groups. All injection methods had less postpolymerization increase in vertical dimension of occlusion (0.63 to 0.41 mm) than the conventional Trevalon control (0.74 mm), but only group 4 was significantly different (P<.004). After storage in water for 28 days, all specimens increased in vertical dimension of occlusion (0.10% to 0.16%) from polymerization techniques, but there were no significant differences between groups. CONCLUSION: Within the limitations of this study, injection molding resulted in a slightly less increase of vertical dimension of occlusion than conventional polymerization techniques, the difference being significant for Microbase compared with the conventional Trevalon control. PMID- 12589285 TI - In vitro kinetic study of absorbency of retraction cords. AB - STATEMENT OF PROBLEM: The soaking time required for liquid uptake by retraction cords is a crucial factor in the successful gingival retraction procedure. PURPOSE: The aim of this investigation was to determine the optimal soaking time for 3 retraction cords of different thickness to ensure adequate uptake of the hemostatic solution. MATERIAL AND METHODS: Braided ULTRAPAC retraction cords of 3 different thicknesses (No. 00, No. 0, and No. 1) with identical lengths (35 mm) were used. The capability of the cords to absorb liquids was measured by a gravimetric method. Different sulcus retraction solutions (epinephrine, aluminum chloride, and ferric sulfate) were tested, with physiological saline solution used as the control. The cords were soaked for various time intervals (2 seconds; 1, 5, and 60 minutes; and 24 hours) in the medicament solutions at room temperature. Immediately before immersion, air inclusions that may have inhibited the inner moistening of the cords were manually pressed out. In each of the 4 treatment groups, 75 pieces of cord were tested by determining the grams of fluid absorbed by grams of dry cord (gram/gram). Before the cord was weighed, excess fluid accumulating on the outer surface of the cord was removed by filter paper saturated in the corresponding test solution. The data were analyzed by F test analysis, and P<.05 was regarded as significant. RESULTS: In each group, regardless of the cord type (No. 00, No. 0, or No. 1) or medicament solutions tested, the amount of fluid absorbed increased with the soaking time, but to different extents. A logarithmic relationship in a linear plot between the amount of fluid absorbed and the soaking time was demonstrated. The relationship established offered an exact determination of both the rate and the saturation level of liquid uptake. The rate of liquid uptake calculated from the saturation equations exhibited significant correlation with the cord thickness (P<.05). The saturation levels of the solutions did not show correlation with the cord thickness (P>.30). CONCLUSION: Within the limitations of this study, the results indicated that 20 minutes of soaking time was necessary for saturation of the cords before use, provided that air trapped within the cords was removed. In addition to the soaking time, the saturation of the cords with the solutions largely depended on the wetting of the cords. PMID- 12589286 TI - Intrarater repeatability of shade selections with two shade guides. AB - STATEMENT OF PROBLEM: Color research has shown that shade guides do not always represent the color of natural teeth. Moreover, visual evaluation has been found to be unreliable and inconsistent. PURPOSE: This investigation evaluated the effects of 2 shade guides on the intrarater repeatability (reliability) of prosthodontists and general practitioners with regard to shade selection. MATERIAL AND METHODS: Ten prosthodontists and ten general practitioners (all men, 35-45 years old) with an average practice experience of 14 years participated in this study. Examiners were tested to eliminate color blindness. Each clinician used Vita Lumin Vacuum and Vitapan 3D-Master shade guides to determine the shades of the maxillary right canines of 20 patients following a standard protocol. The identification codes of the shade tabs were masked to prevent shade memory. All teeth were polished before each shade selection, and the selection process was standardized for controlled lighting and procedures. Shade selections were randomly repeated 1 month later by the same practitioners on the same group of patients in accordance with the same shade-selection protocol. Analysis of variance and t tests for individual comparisons among means were performed (P<.05). RESULTS: Significant interactions were found between the effects of shade guide system and specialty training on intrarater repeatability (P<.0001, analysis of variance). The intrarater repeatability of prosthodontists was significantly higher than that of general practitioners when the Vita Lumin Vacuum shade guide was used (P<.0001, t test). Use of the Vitapan 3D-Master shade guide significantly improved the intrarater repeatability of general practitioners compared with the Vita Lumin Vacuum shade guide (P<.0005). This improvement was not significant, however, among prosthodontists (P=.2861). CONCLUSION: Within the limitations of this study, the prosthodontists demonstrated superior intrarater repeatability in shade selection, especially when the Vita Lumin Vacuum shade guide was used. Use of the Vitapan 3D-Master shade guide notably improved intrarater repeatability among the general practitioners. PMID- 12589287 TI - Comparison of condylar guidance setting obtained from a wax record versus an extraoral tracing: a pilot study. AB - STATEMENT OF PROBLEM: Protrusive condylar angles can be determined by use of wax protrusive records (WPR), pantographic tracings (PT), and occlusal wear facets. These methods generate different angles in the same patient. PURPOSE: The purpose of this study was to compare the condylar inclination angles found by use of the WPR in a Hanau articulator with those found by use of the Whip-Mix PT quick-set recorder. The occlusal wear facet method was not part of this study. MATERIAL AND METHODS: Ten subjects were chosen at random from a dental school patient population. All subjects had a majority of maxillary and mandibular teeth present and according to the examination criteria used in the school's Division of Occlusion, they were healthy with no signs/symptoms of temporomandibular disorders. The WPR was accepted when it was evident that the patient had protruded straight forward at least 6 mm anterior to centric relation, as shown on the Hanau articulator by the condylar spheres having moved anteriorly an equal distance of 6 mm on both sides. The condylar inclination on the articulator was adjusted accordingly. The PT was used according to the manufacturer's directions, which included the use of an intraoral clutch adapted to the mandibular arch, connected to an extraoral facebow with bilateral inscribing pointers. A holder frame was adapted to the face with bilateral flags where the inscribing pointers traced the protrusive movement of the jaw. All the readings were in degrees. No control was used in this project. To test whether there was a significant difference between the 2 independent samples, a Mann-Whitney U test was performed (P=.01). RESULTS: The mean results for the WPR were as follow: right side (28.1 degrees, SD 8.94); left side (31.50 degrees, SD 9.73). For the PT, results were: right side (41.10 degrees, SD 7.53); left side (35.5 degrees, SD 6.43). When right and left side protrusive condylar angles were combined, the values were: WPR (29.80 degrees, SD 9.25); PT (38.30 degrees, SD 6.98). The differences on the right condylar values were significant (P=.01). There was no statistical difference on the left side. CONCLUSIONS: Within the limitations of this study, it was concluded that the PT technique yielded greater values for the protrusive condylar inclination than the WPR. PMID- 12589288 TI - Effect of the silicone disclosing procedure on the shear bond strength of composite cements to ceramic restorations. AB - STATEMENT OF PROBLEM: There is no evidence-based information on how ceramic restorations with an adhesive bond between restoration material and composite cement may be influenced by a silicone disclosing agent. PURPOSE: The aim of this study was to determine the effects of the silicone disclosing procedure on the shear bond strength of composite cements in the luting of industrial sintered and laboratory sintered ceramic restorations. MATERIAL AND METHODS: Thirty standardized (15 x 10 x 9 mm) prefabricated ceramic specimens (Groups 1, 3, 5) and 30 standardized (15 x10 x 9 mm) conventionally sintered ceramic specimens (Groups 2, 4, 6) were roughened with sandpaper (800-grit). Each group contained 10 specimens. Groups 3 and 4 were conditioned with hydrofluoric acid and primed with silane solution after the use of a silicone disclosing procedure. Groups 1 and 2 served as the control groups, where no silicone disclosing procedure was performed. Groups 5 and 6 were insulated with glycerine before the silicone disclosing procedure. A glass tube (4.5 mm in diameter) was used to apply a cylinder of dual-polymerized composite cement to the conditioned surfaces. All specimens were submitted to 5000 thermocycles (5 degrees to 55 degrees C) to simulate the in vivo situation. The specimens were subjected to a shear-pull test at a constant crosshead speed of 5 mm/min with a universal testing machine. The comparative shear bond strengths were analyzed by use of Duncan's test (alpha=0.05). RESULTS: Shear bond strength values for Groups 1 (9.86 +/- 4.97 MPa) and 2 (9.56 +/- 4.47 Mpa) were obtained with no significant differences. Lower but significantly undifferent values were obtained for Groups 3 (7.49 +/- 4.67 MPa) and 4 (7.62 +/- 3.49 MPa) after the use of a silicone disclosing procedure. In Groups 5 (8.21 +/- 4.75 MPa) and 6 (8.22 +/- 3.59 MPa), including insulation with glycerine before the silicone disclosing procedure, no significant differences were obtained. CONCLUSION: Within the limitations of this study, the use of silicone disclosing procedures before conditioning the ceramic surface did not lead to a significant reduction of the shear bond strength between ceramic and composite cement. The ceramic materials used (industrial sintered versus laboratory-sintered ceramic) had no significant influence on adhesion. PMID- 12589289 TI - Orthodontic bonding to porcelain: a comparison of bonding systems. AB - STATEMENT OF PROBLEM: Direct bonding of orthodontic brackets to porcelain surfaces has been plagued by failure. PURPOSE: The purpose of this study was to compare the bond strengths of several different bonding systems when bonding orthodontic brackets to porcelain-fused-to-metal surfaces. MATERIAL AND METHODS: Fifty natural glazed feldspathic porcelain-fused-to-noble metal disks 6 mm in diameter and 3 mm in height (1 mm metal and 2 mm porcelain) were fabricated and divided into 5 groups of 10. A different bonding system (GC America Fuji LC, American Ortho Spectrum, 3M Transbond, TP Orthodontics Python, and Kerr Herculite) was assigned to each group, and 50 identical orthodontic brackets were bonded (with the above mentioned systems) to each disk according to each manufacturer's instructions. Each system except TP Orthodontics Python conditioned with phosphoric acid (35% to 37.5%) and all systems were primed with silane before bonding. The specimens were subjected to gradual shear forces up to 123 N in a universal testing machine (Instron Corp, Canton, Mass.) until fracture. The shear bond strength of the bonding systems between the porcelain surface and the bracket was measured in megapascals (MPa). Failures were observed via a Zeiss optical microscope (10x); Tukey's HSD Test and analysis of variance were used to determine significance between the bonding systems at P<.05 level of significance. RESULTS: Failure of all of specimens was adhesive between the porcelain surface and the bonding agents. On the basis of a current literature review, bonding systems were categorized as clinically acceptable if they had a shear bond strength of 6 to 8 MPa. The 3M Transbond Bonding System, American Orthodontics Spectrum Bonding System, and GC America Fuji Ortho LC Bonding System performed within this clinically acceptable range (6 to 8 MPa), whereas Kerr Herculite Bonding System and TP Orthodontics Python Bonding System did not (2 to 4 MPa). The bond strengths of GC America Fuji Ortho LC, 3M Transbond, and American Orthodontics Spectrum were significantly greater (mean = 2.3 times) than TP Orthodontics Python or Kerr Herculite bonding systems. CONCLUSION: Within the limitations of this study, the results reaffirm the regimen of conditioning with phosphoric acid and priming with silane before bonding orthodontic brackets to feldspathic porcelain fused to noble metal. All products indicated for this purpose may not achieve satisfactory bond strengths; however, because they do not all include the critical steps of conditioning with phosphoric acid and priming with silane. The 3M Transbond Bonding System, American Orthodontics Spectrum Bonding System, and GC America Fuji Ortho LC Bonding System performed within the clinically acceptable range (6 to 8 MPa), whereas Kerr Herculite Bonding System and TP Orthodontics Python Bonding System did not (2 to 4 MPa). PMID- 12589290 TI - Effect of a fluoride varnish on the margin leakage and retention of luted provisional crowns. AB - STATEMENT OF PROBLEM: Provisional crowns cemented with provisional luting agents are susceptible to washout, margin leakage, and secondary caries when placed for a prolonged period. PURPOSE: This study was conducted to determine the effect of combining a varnish containing 2.26% NaF with 2 provisional luting agents on the margin leakage and retention of provisional crowns. MATERIAL AND METHODS: Acrylic resin provisional crowns were fabricated for 8 shoulder-prepared molars. The eight provisional (N=24) crowns were luted individually with Temp-Bond (TB), Freegenol (FG), or Duraphat (DU). Specimens were thermocycled 500 times (5 degrees and 60 degrees C) with a 1-minute dwell time, stored in 100% relative humidity at 37 degrees C for 6 days, and then immersed in a 0.5% Gentian violet solution for 24 hours. Seven days after cementation, a removal test of the crowns (shear retention test) was conducted with a universal testing machine at a crosshead speed of 5 mm/min. Retention was determined as the maximum recorded force needed for crown dislodgment. DU varnish was applied to the inner surface of the dislodged crowns with no removal of the cement layer TB, FG (N=16). The crowns were relined with a 0.5-mm layer of acrylic resin and luted with a combination of luting agent and DU (TB, FG) N=16. No luting agent (NC) served as the control (N=8). Results were analyzed with the Wilcoxon matched-pairs signed ranks test. Leakage at the margins was assessed with a 4-level dye penetration scale, and statistical differences were identified with a chi(2) test. All hypothesis testing was conducted at the 95% level of confidence. RESULTS: The mean 7-day retention forces were as follows: 44.5 N (Temp-Bond), 51.6 N (Freegenol), and 35.9 N (Duraphat). There were no significant differences among these values. Duraphat combined with Freegenol decreased the retention of provisional crowns, but Duraphat combined with Temp-Bond increased the retention of provisional crowns by 69-145%. Duraphat alone and in combination with both provisional luting agents significantly reduced margin leakage (P<.05). The least margin leakage was evident when the provisional crowns were luted with Duraphat alone. CONCLUSION: With regard to retention and margin leakage, the results of this study suggest that Duraphat varnish can be successfully used as provisional luting agent for single provisional crowns. PMID- 12589291 TI - A new method for recording and reproducing cast orientation on a survey table. AB - Cast position on a survey table is determined with an inclinometer, allowing tilt to be measured in 1-degree increments in the frontal and sagittal planes. With this device, cast reorientation is simplified and tilt modifications can be accomplished accurately because of direct visualization of tilt measurements in the 2 planes. PMID- 12589292 TI - Metal framework modifications to accommodate wrought wire clasps in distal extension removable partial dentures. AB - The article discusses minor modifications of the metal framework of distal extension removable partial dentures to accommodate the wrought wire assembly. PMID- 12589293 TI - Assessing the space limitation inside a complete denture for implant attachments. AB - Accurate determination of the space defined by the physiological contouring of a complete denture (before and after implant surgery) is required during the treatment-planning phase for an implant overdenture. This determination assists in formulating the proper surgical treatment plan and in selecting the appropriate independent implant overdenture attachment. This article presents a technique that provides clear, visual space limitation analysis of the removable prosthesis. The described method allows the clinician to determine the available space before the implant or attachment is placed. PMID- 12589294 TI - Composite indirect-direct method for fabricating multiple-unit provisional restorations. AB - This article describes a method for fabricating multiple-unit provisional restorations by use of the indirect-direct method without the need for abutment preparation on diagnostic casts. Composite is used with fiber reinforcement to create a provisional shell in a vinyl polysiloxane matrix. This method reduces chair time, as well as laboratory time required to fabricate provisional restorations. Control of restoration contours and incisal edge position is also enhanced. PMID- 12589295 TI - A crown venting technique for the cementation of implant-supported crowns. AB - With the increased use of implant systems for the replacement of single teeth, some dental practitioners are choosing to permanently cement the final restoration instead of using the screw-retention modality. Many of these restorations have subgingival margins; therefore, the cementation technique becomes a critical procedure because incomplete seating of the restoration or excess cement can be lodged in the gingival sulcus. The use of a cement escape way or venting technique for the cementation of an implant-supported restoration is described. PMID- 12589296 TI - Technique for repair of multiple abutment teeth under preexisting crowns. AB - This technique describes a simple procedure for repairing multiple fractured or broken-down abutment teeth under existing crowns. The technique uses a Teflon tape polytetrafluoroethylene (PTFE) as a matrix and separating medium for fabrication of an autopolymerizing composite core material with digital pressure. This technique enables the dentist to solve an emergency clinical situation in a relatively short amount of time, pending further restoration with endodontic treatment and a cast post and core procedure. PMID- 12589297 TI - Special technique for attachment incorporation with an implant overdenture. AB - The Hader bar system is a popular bar and clip concept because of its 20-degree clip rotation, simplicity, and versatility. Two procedures may be used to incorporate the Hader nylon clip into a denture base. The direct technique consists of attaching the clip to the denture base as a clinical procedure. With the indirect technique, the clip is attached during laboratory processing. An alternative method of attaching the clip with a metal superstructure is presented. This procedure combines the advantages of the direct and indirect techniques for incorporation of the nylon clip of the Hader bar into the denture base. PMID- 12589298 TI - An implant-retained auricular impression technique to minimize soft tissue distortion. AB - Achieving adaptation of an auricular prosthesis begins with an accurate impression. It is important to consider how the selection of the impression materials will affect the final outcome of the prosthesis. A procedure is presented to minimize the distortion of the soft tissues caused by the impression materials and procedure. The procedure consists of splinting the implant impression copings, then recording the soft tissue in silicone impression material, followed by the application of acrylic tray resin to provide rigidity. PMID- 12589300 TI - Length of normal labor in women of Hispanic origin. AB - Emanuel Friedman in the 1950s established means and statistical guidelines for normal lengths of labor. The childbearing population in the United States has changed considerably since Friedman's research was conducted. This study documented the duration of labor in a cohort of 240 Hispanic women who had normal vaginal births of singleton term infants from January 1995 through December 1998 and compared these results with the mean duration of the first and second stages of labor as established by Friedman. The mean duration of the active phase first stage labor duration for nulliparous Hispanic women was 6.2 hours, and for multiparous Hispanic women was 4.4 hours, both significantly longer than Friedman's group (P <.01). The mean duration of the second stage of labor in nulliparous Hispanic women was 54.2 minutes and for multiparous Hispanic women was 22.2 minutes, not significantly different from Friedman's group (P =.5 and P =.09, respectively). PMID- 12589301 TI - Where is the "E" in MCH? The need for an evidence-based approach in safe motherhood. AB - Measuring the impact of obstetric interventions on maternal mortality and/or morbidity is especially difficult in developing countries, where most maternal deaths occur. Therefore, program planning has been based on theory rather than proved effectiveness. After reviewing both the strategies that have been promoted to reduce maternal mortality and the adequacy of existing evidence used to justify their selection, the investigators highlight reasons why rigorous criteria for the selection and evaluation of interventions should be adopted. Adequate evaluation of intervention effectiveness under real-life conditions in developing countries is an efficient way to identify interventions for large scale program replication and could speed progress in reducing maternal deaths. PMID- 12589302 TI - Diethylstilbestrol (DES) update: recommendations for the identification and management of DES-exposed individuals. AB - Diethylstilbestrol (DES) was etiologically linked to clear cell adenocarcinoma of the vagina in 1971. This article reviews on-going research and emerging information relevant to DES-related health risks, thereby enabling women's health care providers to maintain an evidence-based practice for their DES-exposed patients. To accomplish these goals, the Center for Disease Control and Prevention (CDC) has initiated a national education campaign. This article describes the reasons for this new initiative, the target audiences, the DES historical framework (including major studies and findings), and populations that are affected. Clinical steps for the identification and management of the DES exposed individual and resultant implications for midwifery and women's health practices are reviewed. PMID- 12589303 TI - Lead poisoning in pregnancy: a case study with implications for midwives. AB - Lead poisoning remains a significant problem in the United States affecting the health of women and children. Although the damage is greatest at higher levels of accumulation, no level of lead has been found to be safe. Over the last 25 years, the blood level of lead thought to be associated with toxicity has dropped dramatically, from to 60 mcg/dL in 1960 to 10 mcg/dL today. Studies confirm that exposure to lead causes kidney damage, encephalopathy, and impaired cognitive function in children and in adults. Recent evidence indicates children with levels less than 10 mcg/dL may suffer from compromised development and intellectual performance later in life. This article discusses the case of a woman found to have lead poisoning during pregnancy. Environmental sources of lead, implications for the immediate and future health of the fetus and mother, and techniques clinicians can use in their practice to minimize the adverse effect of lead on their clients are reviewed. PMID- 12589304 TI - Pharmacotherapeutics for osteoporosis prevention and treatment. AB - Osteoporosis is a silent disease that affects 10 million Americans; 80% of those affected are women. Although the disease is more common in postmenopausal Caucasian women, all ages and races are at risk. Osteoporosis can be a debilitating disease that can cause pain, fractures, depression, and social withdrawal. Signs of osteoporosis include kyphosis, loss of height, and protrusion of the abdomen. Because symptoms generally do not occur until after the disease has progressed, clinicians should include osteoporosis screening and preventative education as part of the regular gynecologic care. Diagnosis is typically made by a dual energy x-ray absorptiometry (DEXA) scan. Treatment consists of dietary and lifestyle changes, along with pharmacologic intervention. Although hormone therapy has been shown to be effective in preventing osteoporosis, the risks of long-term treatment with HRT are discussed. The following effective treatment options for women who have been diagnosed with the disease are discussed: bisphosphonates, calcitonin, and selective estrogen receptor modulators (SERMs). Because midwives regularly care for women of all ages, they are ideal candidates to provide women with preventative education, screening, counseling, and treatment. PMID- 12589305 TI - Changes in postpartum perineal muscle function in women with and without episiotomies. AB - This report presents results of a comparison perineal muscle function between antepartum and postpartum measurements in a cohort of women with different perineal conditions after childbirth. Data were obtained by using prospective electromyographic perineometry measurements to objectively determine perineal muscle function before and after delivery in 102 women. In addition, 24 nulliparous, non-pregnant women were studied to determine the effect of pregnancy on perineal muscle function. Pregnancy is associated with a decrease in perineal muscle strength and endurance compared with the postpartum state. The degree to which women improved or did not improve perineal muscle function after birth was related to perineal trauma at delivery. After controlling for parity, maternal age, birthweight, smoking status, and antepartum scores, the order of best to worst performance was cesarean birth, intact perineum, first-degree perineal injury, second- or third-degree perineal injury, and episiotomy. Pre- and post delivery scores were compared for each woman and analyzed according to perineal outcome. Although all other perineal outcome groups increased muscle function by 6 months postpartum, women with an episiotomy had a mean net loss of perineal muscle performance after birth. These observations do not support the use of episiotomy for the purpose of preserving perineal muscle function. PMID- 12589306 TI - Early diagnosis and treatment of genuine stress urinary incontinence in women after pregnancy: midwives as detectives. AB - Genuine stress incontinence is often a hidden problem in that many women suffering from genuine stress incontinence after delivery do not seek medical advice. This article reviews signs and symptoms of genuine stress incontinence that, when identified, may enable midwives to initiate or suggest treatment as needed. A comprehensive literature search was performed in relevant medical databases. The following adverse risk factors for the development of genuine stress incontinence are vaginal delivery, multiparity, and obesity, with an increased risk for women who did not actively exert pelvic floor training. Prolonged second stage of labor and heavier babies were two factors associated with an increase in the risk of damage to the pelvic floor innervation as well as genuine stress incontinence. Several conservative treatment options, such as special instructions for pelvic floor training, the use of weight cones, and electrical stimulation, are reported to alleviate genuine stress incontinence symptoms. PMID- 12589307 TI - An unexpected adverse drug effect. AB - Adverse drug-drug interactions can occur between active and/or inactive ingredients in different formulations. The occurrence of a disulfiram reaction that developed postpartum following a 7-day course of metronidazole is presented. The case is presented, followed by a discussion of the mechanism of action and treatment. Recommendations for prevention of adverse drug effects are reviewed. PMID- 12589309 TI - Share with women. PMID- 12589315 TI - Art and science of midwifery. PMID- 12589318 TI - Are health plans changing their views on nutrition service coverage? PMID- 12589319 TI - A picture is worth a thousand words: digital photography and diet assessment. PMID- 12589320 TI - Good reading. PMID- 12589321 TI - A ripple that became a tsunami. PMID- 12589322 TI - The need for a nutrition-related quality-of-life measure. PMID- 12589323 TI - Folate intakes and awareness of folate to prevent neural tube defects: a survey of women living in Vancouver, Canada. AB - OBJECTIVE: To estimate folate intake and knowledge in women of childbearing age, in relation to risk of neural tube defects. SUBJECTS/SETTING: One hundred forty eight women (aged 18 to 45 years) in the Vancouver area of British Columbia, Canada. DESIGN: Using an interviewer-administered survey, we examined women for folate knowledge and the relation of folate knowledge to intake in a random sample. Contribution of folate from food, fortified grain products, and supplements was assessed by validated semiquantitative food frequency questionnaire. STATISTICAL ANALYSIS PERFORMED: Descriptive statistics, t tests, Chi;(2), Pearson correlation analysis. RESULTS: Mean daily folate intake from food, fortified foods, and supplementation was 812+/-710 Dietary Folate Equivalents (DFE)/day. Fortification of bread and grain products contributed 104+/-68 microg synthetic folic acid (SFA)/day (equal to 174+/-114 DFE), and supplements contributed 205+/-388 microg SFA/day. Although 86% of women met the Estimated Average Requirement (320 DFE/day) for folate, only 26% met the recommendation (400 microg SFA/day) for women capable of becoming pregnant. Most (95%) of the women had heard of folate, but only 25% knew that it could prevent birth defects. One-fourth of the women had good or very good knowledge of folate rich foods. However, folate knowledge was not related to its intake. The most common sources of folate information were magazines/newspapers, doctors, and television/radio. Lack of awareness of the importance of folate was the most common reason given for choosing not to use folic acid supplements before pregnancy. However, 78% of the women indicated that, with knowledge of the benefits of folate, they would use supplemental folic acid daily to reduce the risk of birth defects. APPLICATIONS/CONCLUSIONS: Educational strategies are required to increase folate awareness among women and to promote the benefits of periconceptional folic acid supplementation. Targeting physicians to educate women on the importance of folate could be a potentially successful route. J PMID- 12589324 TI - Reproducibility and validity of the secondary level School-Based Nutrition Monitoring student questionnaire. AB - OBJECTIVE: To evaluate the reproducibility and validity of the School-Based Nutrition Monitoring (SBNM) secondary level student questionnaire. DESIGN: Reproducibility was evaluated using a test-retest study design by administering the questionnaire on the same day (morning and afternoon, n=254) and, when appropriate, 9 to 14 days apart (n=259). Validity was assessed by comparing foods selected on the questionnaire with foods reported from a single 24-hour recall covering the same referent period (yesterday) in 209 students. SUBJECTS/SETTING: Eighth grade students in middle schools from central Texas were used: male and female, approximately 75% white, for the reproducibility study, with 15% Hispanic, 6% African American; for the validation study, approximately 38% white, 41% Hispanic, and 17% African American. STATISTICAL ANALYSIS: Spearman rank order correlation, kappa statistic, and percentage agreement were used for both reproducibility and the validation. RESULTS: For the reproducibility study, agreement for questions about food intake "yesterday" were 70% to 98%, with kappa statistics ranging from 0.54 to 0.93 and correlations between 0.66 and 0.97. Questions on recent physical activity had high agreement (66% to 89%) as did "usual" physical activity items, weight loss, and food selection behaviors. Nutrition knowledge items showed relatively weaker reliability: agreements ranged from 47% to 92%, with kappa statistics between 0.30 and 0.56. Attitude questions had weaker agreement (50% to 87%), kappa statistics (0.27 to 0.52), and correlations (0.33 to 0.63). For the validation study, correlation coefficients ranged from 0.32 for breads to 0.68 for milk and beans. Percentage agreement ranged from 38% for breads to 89% for gravy. APPLICATIONS/CONCLUSIONS: Most questions on the SBNM secondary questionnaire were found to have acceptable reproducibility, whereas validation of food choice behaviors showed results similar to or better than other dietary assessment instruments for this age group. This questionnaire is a useful epidemiologic tool for surveillance, assessing broad intervention effects among groups or providing needs assessment data on selected nutrition and physical activity-related constructs. PMID- 12589325 TI - Comparison of three methods for the determination of body fat in patients on long term hemodialysis therapy. AB - OBJECTIVE: To compare three simple methods of body composition analysis for the assessment of body fat in patients on long-term hemodialysis therapy. DESIGN: Cross-sectional study using the skinfold thickness, bioelectrical impedance analysis, and near-infrared interactance techniques after a hemodialysis session. SUBJECTS/SETTING: Ninety clinically stable patients (57 male/33 female) undergoing hemodialysis at the Dialysis Unit of the Federal University of Sao Paulo. STATISTICAL ANALYSIS: Analysis of variance, intraclass correlation coefficient, and Bland-Altman plot analysis were used for the comparative analysis between the methods. RESULTS: Body fat measurements obtained by skinfold thickness (13.5+/-6.2 kg) and bioelectrical impedance analysis (13.7+/-6.7 kg) were similar, whereas those measured by near-infrared interactance (11.3+/-5.1 kg) were significantly lower in comparison with skinfold thickness and bioelectrical impedance analysis (P<.001). The strongest intraclass correlation coefficient was found between bioelectrical impedance analysis and skinfold thickness (r=0.87), and near-infrared interactance vs skinfold thickness and bioelectrical impedance analysis methods yielded r=0.78 and r=0.76, respectively. Near-infrared interactance showed a progressive underestimation of body fat values in comparison with the bioelectrical impedance analysis technique in patients with higher amount of adiposity. CONCLUSION: In our study, we cannot consider that one method of body composition analysis is more accurate than the other because we did not apply a gold standard method. However, the most simple, long-established, and inexpensive method of skinfold thickness seems to be still very useful to the dietitians' routine for assessing body fat in patients on long term hemodialysis therapy. PMID- 12589326 TI - Heterogeneity in the prevalence of methylenetetrahydrofolate reductase gene polymorphisms in women of different ethnic groups. AB - OBJECTIVE: To determine the prevalence of methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms in women of different ethnic groups and to relate these common mutations to plasma homocysteine, red cell folate, and serum folate. DESIGN: A one-time fasting blood sample was obtained for MTHFR genotype (C677T and A1298C) determinations (n=433). Serum folate, red cell folate, and homocysteine analyses were performed in nonfolic acid supplement users (n=215). SUBJECTS/SETTING: This study involved 433 women from four ethnic groups, including 193 Hispanic women of Mexican descent, 139 white women, 53 Asian women of mixed descent, and 48 African American women. STATISTICAL ANALYSIS PREFORMED: Chi;(2), t Test, and analysis of variance were used. RESULTS: Mexican women (18.1%) had a higher frequency of the 677 TT genotype compared with white (7.2%), Asian (3.8%), and African American (0%) women. White women (7.9%) had a higher frequency of the 1298 CC genotype than the other ethnic groups (range=1.9% to 2.6%). The frequency of compound heterozygosity (677 CT + 1298 AC) was higher in Mexican (17.6%) and white (15.1%) women than Asian and African American ( approximately 4% to 6%) women. In the era of folic acid fortification, neither genotype, independently or together, was associated with homocysteine or blood folate concentrations when ethnic groups were combined. In Mexican women, however, a linear trend (P or =0.016 mg/kg of IgE [in international unit per milliliter] per 4 weeks) in 525 adults with severe allergic asthma. A 16-week steroid-stable phase was followed by a 12-week steroid reduction phase and a 24-week double-blind extension phase. The effect of treatment on asthma-related QOL was evaluated by using the Asthma Quality of Life Questionnaire (AQLQ) administered at baseline and at weeks 16, 28, and 52. RESULTS: The 2 treatment groups were comparable in terms of baseline AQLQ scores. At weeks 16, 28, and 52, omalizumab-treated patients demonstrated statistically significant improvements across all AQLQ domains, as well as in overall score. Moreover, a greater proportion of patients receiving omalizumab achieved a clinically meaningful improvement in asthma-related QOL during each phase of the study. Greater than 50% of both patients and investigators rated treatment similarly with omalizumab as excellent or good compared with less than 40% of placebo recipients. CONCLUSION: In patients requiring moderate-to-high doses of ICSs for severe allergic asthma, the measurably improved disease control afforded by add-on omalizumab therapy is paralleled by clinically meaningful improvements in asthma-related QOL. PMID- 12589346 TI - Allergen detection from 11 fungal species before and after germination. AB - BACKGROUND: Allergens dispersed by airborne fungal spores play an important but poorly understood role in the underlying cause and exacerbation of asthma. Previous studies suggest that spores of Alternaria and Aspergillus release greater quantities of allergen after germination than before germination. It is unknown whether this is true of other allergenic fungi. OBJECTIVE: Our purpose was to investigate the release of allergen from a range of individual fungal spores before and after germination. METHODS: Allergen expression from spores of Alternaria alternata, Cladosporium herbarum, Aspergillus fumigatus, Botrytis cinerea, Epicoccum nigrum, Exserohilum rostratum, Penicillium chrysogenum, Stemphylium botryosum, Curvularia lunata, Trichoderma viride, and Bipolaris spicifera was examined by halogen immunoassays through the use of pooled serum IgE from patients allergic to fungus. Spores were deposited onto protein-binding membranes direct from culture. To germinate spores, samples were incubated in high humidity at room temperature for 48 hours. Ungerminated and germinated samples were then laminated with an adhesive film and immunostained by the halogen assay. The samples were examined by light microscopy, and positive counts (haloed particles) were expressed as percentages of total spores. RESULTS: For 9 of 11 species, between 5.7% and 92% of spores released allergen before germination. Spores of Penicillium and Trichoderma did not release detectable allergen. After germination, all spores that germinated had allergen elution from their hyphae. Eight of 11 species showed a significant increase (P <.05) in the percentage of spores eluting detectable allergen. Localization of allergen along the hyphae varied with species, such that some eluted allergen mainly from hyphal tips and septal junctions whereas others eluted allergen along the entire length. CONCLUSIONS: Increased elution of allergen after germination might be a common feature of many species of allergenic fungi. Although allergens from both spores and hyphae were recognized by human IgE, the extent to which human exposure occurs to allergens eluted from inhaled spores or from hyphae that germinate after deposition in the respiratory tract remains to be explored. The patterns of allergen expression might affect the clinical response to such exposure. PMID- 12589347 TI - Cities as harbingers of climate change: common ragweed, urbanization, and public health. AB - BACKGROUND: Although controlled laboratory experiments have been conducted to demonstrate the sensitivity of allergenic pollen production to future climatic change (ie, increased CO(2) and temperature), no in situ data are available. OBJECTIVE: The purpose of this investigation was to assess, under realistic conditions, the impact of climatic change on pollen production of common ragweed, a ubiquitous weed occurring in disturbed sites and the principal source of pollen associated with seasonal allergenic rhinitis. METHODS: We used an existing temperature/CO(2) gradient between urban and rural areas to examine the quantitative and qualitative aspects of ragweed growth and pollen production. RESULTS: For 2000 and 2001, average daily (24-hour) values of CO(2) concentration and air temperature within an urban environment were 30% to 31% and 1.8 degrees to 2.0 degrees C (3.4 degrees to 3.6 degrees F) higher than those at a rural site. This result is consistent with most global change scenarios. Ragweed grew faster, flowered earlier, and produced significantly greater above-ground biomass and ragweed pollen at urban locations than at rural locations. CONCLUSIONS: Here we show that 2 aspects of future global environmental change, air temperature and atmospheric CO(2), are already significantly higher in urban relative to rural areas. In general, we show that regional urbanization-induced temperature/CO(2) increases similar to those associated with projected global climatic change might already have public health consequences; we suggest that urbanization, per se, might provide a low-cost alternative to current experimental methods evaluating plant responses to climate change. PMID- 12589348 TI - Direct costs of allergic rhinitis in the United States: estimates from the 1996 Medical Expenditure Panel Survey. AB - BACKGROUND: Previous estimates of the cost of allergic rhinitis predate the substantial increase in the use of second-generation antihistamines and intranasal corticosteroids. OBJECTIVE: We sought to update estimates of the direct costs of allergic rhinitis in the United States and to estimate prescription medication expenditures by type of insurance coverage. METHODS: Data from the 1996 Medical Expenditure Panel Survey were used in a cross-sectional analysis of resource use and costs. RESULTS: Approximately 7.7% of the population are estimated to have had allergic rhinitis in 1996. The total direct medical cost of allergic rhinitis was estimated at $3.4 billion, with the majority attributable to prescription medications (46.6%) and outpatient visits (51.9%). Fifty-one percent of the prescription medication expenditures were for second generation antihistamines, 25% for intranasal corticosteroids, and 5% for first generation antihistamines. Fifty-eight percent of patients with allergic rhinitis received 1 or more prescription drugs for its treatment during the study year. Among these patients, mean prescription expenditures were $131 (95% CI, $119 $143), of which $50 (95% CI, $43-$56) were paid out of pocket. The mean prescription medication expenditure was $103 (95% CI, $70-$136) for persons with Medicaid, $155 (95% CI, $140-$169) for private insurance, $213 (95% CI, $0-$521) for other insurance, and $69 (95% CI, $57-$80) for no prescription drug insurance. CONCLUSION: The direct costs of allergic rhinitis have increased substantially since the introduction of second-generation antihistamines and intranasal corticosteroids, especially costs attributable to prescription medications. Individuals with no insurance coverage have higher total out-of pocket prescription expenditures than those with coverage. PMID- 12589349 TI - Exercise-induced bronchoconstriction in children: montelukast attenuates the immediate-phase and late-phase responses. AB - BACKGROUND: Montelukast, a leukotriene receptor antagonist, attenuates exercise induced bronchoconstriction. We and others have shown that there is a late-phase response 3 to 8 hours after exercise in a subset of asthmatic patients. OBJECTIVE: We sought to evaluate the protective effect of montelukast on immediate-phase and late-phase responses after exercise challenges. METHODS: Twenty-two atopic asthmatic children aged 7 to 16 years with reproducible exercise-induced bronchoconstriction (minimum of 15% decrease of FEV(1) from baseline) were enrolled in this placebo-controlled crossover study. Exercise challenges were performed while breathing cold dry air, and FEV(1) measurements were taken up to 480 minutes after exercise. Patients underwent exercise challenges on a screening day and 1 week after placebo treatment. Subsequently, after a week with no treatment, pulmonary function was assessed after breathing dry cold air (control day). Finally, an exercise challenge was carried out after a week of treatment with montelukast. RESULTS: Reproducible late-phase reactions occurred in 5 of 22 patients, which correlated with the extent of the immediate response (P <.05). After 1 week of treatment with montelukast, a significant decrease of immediate responses was observed. Montelukast treatment compared with placebo was associated with a lower mean maximum decrease of FEV(1) (mean +/- SEM: 17.3% +/- 2.4% and 35.1% +/- 2.6%, respectively), decrease of the area above the curve (267.8% +/- 42.7%/min and 868.0% +/- 103.8%/min, respectively), and shorter time for recovery (6.9 +/- 1.1 minutes and 30.9 +/- 4.0 minutes, respectively; P <.05). Treatment with montelukast also abolished late-phase responses. CONCLUSION: Once daily treatment with oral montelukast attenuated the immediate-phase response and abolished the late-phase response induced by means of exercise challenge in asthmatic children. PMID- 12589350 TI - Circadian variation of sputum inflammatory cells in mild asthma. AB - BACKGROUND: Asthma, like many conditions, demonstrates a circadian rhythm with a worsening of lung function in the early morning hours compared with in the late afternoon. OBJECTIVE: Because eosinophilic airway inflammation is a proposed mechanism for worsening asthma, we characterized circadian variation in airway eosinophils and determined its relationship to variability in airway function. METHODS: Pulmonary function testing, sputum induction, and phlebotomy were performed at 7 am and 4 pm in 11 allergic subjects with mild asthma. Sputum was analyzed for cell viability, differential, and eosinophil-derived neurotoxin levels. IL-5 levels in serum were measured by means of ELISA. RESULTS: Subjects had a significant decrease in FEV(1) (median [interquartile range] = 80% [70% 86%] vs 85% [82%-94%], P =.009) and a greater beta-agonist reversibility (median [interquartile range] = 13% [7%-32%] vs 8% [5%-14%], P =.024) in the early morning compared with in the late afternoon. Sputum analysis showed an increase in early morning total sputum leukocytes (median [interquartile range] = 4.3 x 10(6) [2.3 x 10(6) to 6.1 x 10(6)] vs 2.6 x 10(6) [1.7 x 10(6) to 3.6 x 10(6)], P =.044) and eosinophils (median [interquartile range] = 7.0 x 10(4) [2.7 x 10(4) to 18.7 x 10(4)] vs 3.6 x 10(4) [1.0 x 10(4) to 8.2 x 10(4)], P =.024). Furthermore, sputum eosinophils correlated with beta-agonist reversibility (R (s) = 0.665, P =.019). Finally, levels of IL-5 in serum and eosinophil-derived neurotoxin in sputum were significantly increased at 7 am. CONCLUSION: These data suggest that circadian variability in pulmonary function in asthma could be related to changes in airway eosinophil recruitment and activation. PMID- 12589351 TI - Effect of nasal triamcinolone acetonide on lower airway inflammatory markers in patients with allergic rhinitis. AB - BACKGROUND: Allergic rhinitis (AR) and asthma are commonly associated, and similar underlying inflammatory processes link both diseases. AR, even in the absence of asthma, is associated with increased levels of exhaled nitric oxide (ENO) and hydrogen peroxide (H(2)O(2)) in exhaled breath condensate, 2 noninvasive markers of lower airway inflammation. OBJECTIVE: We sought to evaluate the effect of treatment with the nasal steroid triamcinolone acetonide on ENO and exhaled H(2)O(2) in subjects with AR. METHODS: We allocated 23 subjects in a randomized, double-blind, parallel-controlled fashion to 4-week treatment with triamcinolone acetonide (220 microg/d) or matching placebo. RESULTS: ENO levels were greater in the subgroup with concomitant asthma (16/23 subjects) and decreased significantly with triamcinolone acetonide treatment in this subgroup of patients in comparison with patients receiving placebo. Breath condensate levels of H(2)O(2) were higher in patients with AR without asthma than in those with asthma but decreased significantly with triamcinolone acetonide treatment in both subgroups. No changes were observed in bronchial hyperresponsiveness, nasal and asthma symptoms, or peak expiratory flow with active treatment or placebo. CONCLUSION: We conclude that treatment of AR with triamcinolone acetonide results in decrease of 2 noninvasive markers of lower airway inflammation, ENO and H(2)O(2), supporting that upper and lower airway inflammation should be seen as a continuum in subjects with AR with and without asthma. ENO might be a more specific marker of the lower airway inflammation present in asthma. PMID- 12589352 TI - Efficacy of liposomal budesonide in experimental asthma. AB - BACKGROUND: Inhaled corticosteroids, such as budesonide, attenuate the inflammatory response in asthma. However, patient noncompliance and side effects of available inhaled corticosteroids limit their use. Liposomes are currently used in medicine to deliver a variety of drugs. OBJECTIVE: The objective of our study was to determine whether weekly therapy with budesonide encapsulated in sterically stabilized (stealth) liposomes would be comparable to daily budesonide therapy in reducing allergic inflammation. METHODS: Ovalbumin-sensitized C57/Black 6 mice received aerosolized (1) budesonide encapsulated in stealth or conventional liposomes, administered weekly, (2) budesonide (without liposomes), administered either daily or weekly, or (3) empty stealth liposomes, administered weekly. All treatment groups were compared with sensitized untreated or unsensitized mice. Histopathologic examination of the lung tissues and measurements of eosinophil peroxidase activity, peripheral blood eosinophil counts, and total serum IgE levels were done weekly for 4 weeks. RESULTS: Weekly therapy with budesonide encapsulated in stealth liposomes was as effective as daily budesonide therapy in decreasing lung inflammation and lowering eosinophil peroxidase activity, peripheral blood eosinophils, and total serum IgE levels. In none of the other groups was there a significant decrease in the inflammatory parameters evaluated. CONCLUSION: We conclude that weekly therapy with budesonide encapsulated in stealth liposomes is as effective as daily budesonide in reducing markers of lung inflammation in experimental asthma. This novel strategy offers an effective alternative to standard daily budesonide therapy in asthma and has the potential to reduce toxicity and improve compliance. PMID- 12589353 TI - Breast-feeding and the prevalence of asthma and wheeze in children: analyses from the Third National Health and Nutrition Examination Survey, 1988-1994. AB - BACKGROUND: Asthma prevalence has increased dramatically in recent years, especially among children. Breast-feeding might protect children against asthma and related conditions (recurrent wheeze), and this protective effect might depend on the duration and exclusivity of the breast-feeding regimen. OBJECTIVE: We sought to determine whether there is an association between breast-feeding and asthma, recurrent wheeze, or both in children up to 72 months of age and whether the duration and exclusivity of breast-feeding affect this association. METHODS: Data were from the third National Health and Nutrition Examination Survey, a nationally representative cross-sectional survey conducted from 1988 to 1994. We tested for significant associations between breast-feeding and physician diagnosed asthma and recurrent wheeze (> or =3 episodes in the past 12 months) before and after adjusting for potential confounders. RESULTS: Crude analyses showed that breast-feeding was associated with significantly reduced risks for asthma and recurrent wheeze in children 2 to 71 months of age, but after adjusting for potential confounders, these overall protective associations attenuated and were no longer statistically significant. However, 2 new and important associations were revealed after adjusting for confounders: (1) compared with never breast-fed children, ever breast-fed children had significantly reduced odds of being diagnosed with asthma and of having recurrent wheeze before 24 months of age, and (2) among children 2 to 71 months of age who had been exposed to environmental tobacco smoke, those who had ever been breast fed had significantly reduced risks of asthma and wheeze compared with those who had never been breast-fed. CONCLUSIONS: Breast-feeding might delay the onset of or actively protect children less than 24 months of age against asthma and recurrent wheeze. Breast-feeding might reduce the prevalence of asthma and recurrent wheeze in children exposed to environmental tobacco smoke. PMID- 12589354 TI - Comparative in vivo bioactivity of modern H1-antihistamines on AMP challenge in atopic asthma. AB - BACKGROUND: Modern H(1)-antihistamines differ in their in vitro binding affinity, but their comparative in vivo bioactivity in asthmatic airways is unknown. OBJECTIVES: We compared clinically recommended doses of 3 H(1)-antihistamines on airway hyperresponsiveness to AMP challenge (the primary outcome variable). METHODS: Sixteen atopic patients with mild-to-moderate asthma of whom 10 were receiving inhaled corticosteroid therapy (all had positive results to house dust mite on skin prick testing) were randomized in a double-blind, placebo controlled, cross-over fashion to receive single doses of 5 mg of desloratadine, 180 mg of fexofenadine hydrochloride (FEX), 5 mg of levocetirizine dihydrochloride (LEV), or placebo, with AMP challenge performed 12 hours after dosing. RESULTS: All H(1)-antihistamines demonstrated significantly greater (P <.05) geometric mean +/- SEM AMP PC(20) values compared with that of placebo (86 +/- 29 mg/mL): desloratadine, 189 +/- 54 mg/mL; FEX, 176 +/- 57 mg/mL; and LEV, 163 +/- 48 mg/mL. Prechallenge forced expiratory flow at 25% to 75% of maximal lung volume (percent predicted) but not FEV(1) was significantly higher (P <.05) for all H(1)-antihistamines compared with that of placebo (53% +/- 4%): desloratadine, 62% +/- 4%; FEX, 62% +/- 4%; and LEV, 59% +/- 3%. There were no significant differences in either AMP PC(20) or lung function values among the H(1)-antihistamines. CONCLUSION: Single doses of H(1)-antihistamines improved airway hyperresponsiveness and small-airways caliber to a similar degree. Data for in vitro binding affinity do not therefore translate into commensurate differences in in vivo bioactivity at clinically recommended doses. PMID- 12589355 TI - Cysteinyl leukotriene expression in chronic hyperplastic sinusitis-nasal polyposis: importance to eosinophilia and asthma. AB - BACKGROUND: Chronic hyperplastic eosinophilic sinusitis (CHS) results from the unregulated proliferation of eosinophils, T(H)2-like lymphocytes, goblet cells, mast cells, and fibroblasts and is present in most patients with asthma. The frequent coexpression of these disorders and their shared pathophysiology suggests that these are similar disorders affecting the upper and lower airways. OBJECTIVE: We evaluated the expression of cysteinyl leukotrienes (CysLTs) in sinus tissue from subjects with CHS compared with that seen in healthy sinus tissue. METHODS: Nasal polyp and sinus tissue was evaluated from 58 individuals undergoing elective functional endoscopic sinus surgery. The diagnosis of CHS was demonstrated through the presence of eosinophilia and activated (EG2(+)) eosinophils, as determined by means of tissue immunohistochemistry. Data were compared with those from both nasal polyp tissue without eosinophilic inflammation and healthy control sinus tissue obtained from the sinus ostiomeatal complex at the time of surgery for unrelated disorders. CysLTs were quantified by means of ELISA in lipid-extracted tissue. Activation of the metabolic pathway leading to CysLT synthesis was demonstrated by ribonuclease protection. Subjects were genotyped for leukotriene C(4) (LTC(4)) synthase C-to-A promoter polymorphism. RESULTS: CysLT concentrations were significantly higher in tissue obtained from subjects with CHS (776.7 +/- 201.9 pg/g tissue) compared with that seen in healthy sinus tissue (355.7 +/- 101.6 pg/g tissue, P <.03). CysLT concentrations within noneosinophilic nasal polyps (328.0 +/- 116.4 pg/g tissue) were similar to those in control tissue. The presence of CysLTs in CHS was associated with increased expression of LTC(4) synthase mRNA. The C-to-A promoter polymorphism was associated with trends toward the increased presence of CHS and CysLTs. CONCLUSIONS: CHS is characterized by the increased presence of CysLTs when compared with concentrations seen in tissue from patients with chronic inflammatory sinusitis or healthy sinus tissue. These studies support the use of LT modifiers as anti-inflammatory agents that might have clinical benefit in patients with these disorders. PMID- 12589356 TI - Identification of grape and wine allergens as an endochitinase 4, a lipid transfer protein, and a thaumatin. AB - BACKGROUND: Few allergic reactions to grape are reported in the literature. In some cases an association with peach and cherry allergy was observed. No IgE mediated reactions to wine have been described, and no grape major allergens have yet been identified. OBJECTIVE: We describe several severe reactions to grape or wine. We characterized the grape major allergens and tried to identify the allergen in wine. METHODS: We collected documented histories of allergic reactions to grape and wine. Grape allergens were identified by means of SDS-PAGE and immunoblotting and purified by means of HPLC. Using amino acid sequencing and mass spectrometry, we identified the family of proteins to which the allergens belong. Cross-reactivity with peach and cherry was evaluated by means of cross wise inhibition experiments. RESULTS: Eleven patients with reactions to grape and 3 with anaphylactic reactions to wine were recruited. The major allergens were an endochitinase 4A and a lipid-transfer protein (LTP) that was homologous to and cross-reactive with peach LTP. A 24-kd protein homologous to the cherry thaumatin like allergen was a minor allergen. Endochitinase 4A is very likely the allergen in vino novello and in vino Fragolino. CONCLUSIONS: Grape and wine might cause severe allergic reactions in sensitive patients. The major allergens of grape are endochitinase 4A, which is also the allergen of wine, and an LTP cross-reacting with the peach major allergen. PMID- 12589357 TI - Corticosteroid-induced apoptosis in mouse airway epithelium: effect in normal airways and after allergen-induced airway inflammation. AB - BACKGROUND: Damage to the airway epithelium is one prominent feature of the damage seen in chronic asthma. Cortico-steroids induce apoptosis in inflammatory cells, which in part explains their ability to suppress airway inflammation. However, corticosteroid therapy does not necessarily reverse the epithelial damage seen in asthmatic airways. OBJECTIVE: We hypothesized that corticosteroids might induce airway epithelial cell apoptosis as one potential explanation for this persistent damage. METHODS: BALB/c mice were treated with 1 mg/kg dexamethasone by means of intraperitoneal injection for 3 days to 4 weeks. Additional mice were sensitized and challenged with ovalbumin and then followed for 14 days with or without dexamethasone treatment starting on day 4 after challenge. Apoptosis was measured by using terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling and immunohistochemistry for the p85 cleavage product of poly (adenosine diphosphate-ribose) polymerase. Shed epithelial cells were counted in bronchoalveolar lavage fluid. RESULTS: In a time dependent manner dexamethasone treatment increased epithelial cell apoptosis and shedding into the airway lumen. This was not associated with any change in the abundance of the apoptotic regulator Bcl-x(L). In animals sensitized and challenged with ovalbumin, treatment with 1 mg/kg dexamethasone starting 4 days after challenge reversed the inflammatory changes but did not reverse either epithelial cell shedding or apoptosis seen after allergen challenge. CONCLUSION: Corticosteroids might induce apoptotic cell death of airway epithelium in vivo and fail to mitigate epithelial cell shedding and apoptosis elicited by means of allergen challenge. This raises the possibility that at least one of the major components of chronic airway damage in asthma, epithelial shedding, might in part result from a major therapy used for disease control. PMID- 12589358 TI - Effect of acute and chronic inflammatory stimuli on expression of protease activated receptors 1 and 2 in alveolar macrophages. AB - BACKGROUND: Protease-activated receptors 1 and 2 (PAR-1 and PAR-2) are 7 transmembrane G protein-coupled receptors activated by serine proteases in many cell types, including monocytes-macrophages, leading to the production of pro inflammatory mediators, cytokines, and growth factors. OBJECTIVE: We determined the influence of chronic smoking and asthma on the expression of PAR-1 and PAR-2 receptors on alveolar macrophages (AMs). METHODS: We used RT-PCR and immunocytochemistry with confocal microscopy to determine mRNA and protein expression of PAR-1 and PAR-2 in AMs obtained from healthy smokers, asthmatic patients, and healthy subjects. In addition, we examined the effect of IL-1beta and LPS. RESULTS: PAR1 mRNA was decreased, whereas PAR2 mRNA was increased in 24 hour cultured AMs from smokers when compared with values in AMs from healthy subjects. Paradoxically, there was a higher degree of PAR-1 protein staining in AMs from smokers, whereas PAR-2 staining was similar in smokers and healthy subjects. PAR-1 and PAR-2 mRNA and protein expression were similar in asthmatic patients and control subjects. IL-1beta and LPS had no effect on PAR1 and PAR2 gene expression by AMs. CONCLUSIONS: There is a dissociation between gene and protein expression of PAR-1 and PAR-2. PAR-1 protein overexpression in AMs from smokers might be important in the pathophysiology of chronic airways disease. PMID- 12589359 TI - Diagnostic paradigm for evaluation of male patients with chronic granulomatous disease, based on the dihydrorhodamine 123 assay. AB - BACKGROUND: Chronic granulomatous disease (CGD) is a phagocyte disorder caused by mutations in nicotinamide dinucleotide phosphate (NADPH) oxidase subunits. The dihydrorhodamine 123 (DHR) assay is an effective test for CGD that for most patients also might help to differentiate between the 2 most common forms, X linked (X) gp91(phox) defect CGD and autosomal recessive (AR) p47(phox) defect CGD. However, some male patients with X-CGD have DHR patterns that overlap the AR CGD pattern. OBJECTIVE: The purpose of this investigation was to develop a diagnostic paradigm to delineate male patients with X-CGD expressing a DHR pattern suggestive of p47(phox) deficiency. METHODS: The DHR assay measured change in fluorescence of DHR-loaded granulocytes after phorbol myristate acetate (PMA) stimulation. Western blot analysis measured the presence of NADPH oxidase subunits gp91(phox), p47(phox), p67(phox), and p22(phox). CYBB exonic sequencing was performed on PCR-amplified genomic DNA through use of intronic flanking primers. Ferricytochrome-c assay evaluated specific superoxide production by PMA stimulated granulocytes. RESULTS: Although 83% of patients with X-CGD have virtually no neutrophil DHR activity, we found that 17% of patients, proven to have X-CGD by other criteria, have modest DHR activity that is most consistent with p47(phox) deficiency. We describe a diagnostic paradigm to deal with such patients, and we present 2 cases, along with results of additional studies, including carrier evaluation, protein assessment, and mutation analysis, that are useful in establishing the genotype under these circumstances. DHR assays from the 2 patients described showed a fluorescence shift most characteristic of p47(phox)-AR-CGD; however, each of the patients' mothers showed mosaicism with a bimodal DHR pattern. Patient 1 had some gp91(phox) protein with a Y41D mutation and modest superoxide activity. Patient 2 had a normal level of gp91(phox) protein with a C537R mutation without detectable superoxide activity, as determined by ferricytochrome-c assay, despite the modest DHR activity. CONCLUSIONS: Evaluation of male patients with CGD with modest DHR activity should initially include evaluation of potential female carriers for mosaicism with the use of the DHR assay. In circumstances in which this is uninformative, patients should be referred to centers capable of additional testing, including Western blot analysis and CYBB mutation analysis, to clarify the disease genotype. PMID- 12589360 TI - The novel synthetic immune response modifier R-848 (Resiquimod) shifts human allergen-specific CD4+ TH2 lymphocytes into IFN-gamma-producing cells. AB - BACKGROUND: In experimental models, imidazoquinolines exhibit several immunomodulatory activities via Toll-like receptor signaling on cells of the innate immunity. OBJECTIVE: The aim of our study was to investigate whether R-848 (Resiquimod), a small-molecular-weight synthetic compound belonging to the imidazoquinoline family and known for its ability to substantially delay the onset of recurrent genital herpes lesions in both animals and human beings, could influence, at least in vitro, the cytokine production profile of human hapten- or allergen-specific T cells. METHODS: Ampicillin- and Der p 1-specific T-cell lines were derived from peripheral blood of allergic donors in the absence or presence of R-848 and assessed by flow cytometry at the single-cell level for their ability to produce IL-4 and/or IFN-gamma. RESULTS: R-848 induced both hapten- and allergen-specific circulating T cells, including T(H)2 effectors, to produce IFN gamma and even to lose the ability to produce IL-4, thus shifting their phenotype of cytokine production to a type 0 (T(H)0) or even T(H)1 profile. This effect was associated with an increase in the production of IL-12, IFN-alpha, IL-18, TNF alpha, IL-10, and IL-15 by CD14(+) cells, as well as an increase in the proportions of IFN-gamma-producing CD3(-)CD16(+) (natural killer) cells. CONCLUSIONS: These results suggest that R-848, and probably other imidazoquinolines, might be used as adjuvants in view of novel allergen-specific immunotherapeutic regimens for the treatment of allergic disorders. PMID- 12589361 TI - Effect of probiotic Lactobacillus strains in children with atopic dermatitis. AB - BACKGROUND: Recent studies suggest that oral bacteriotherapy with probiotics might be useful in the management of atopic dermatitis (AD). OBJECTIVE: The purpose of this investigation was to evaluate the clinical and anti-inflammatory effect of probiotic supplementation in children with AD. METHODS: In a double blind, placebo-controlled, crossover study, 2 probiotic Lactobacillus strains (lyophilized Lactobacillus rhamnosus 19070-2 and Lactobacillus reuteri DSM 122460) were given in combination for 6 weeks to 1- to 13-year-old children with AD. The patients' evaluations were registered after each intervention (ie, better, unchanged, or worse). The clinical severity of the eczema was evaluated by using the scoring atopic dermatitis (SCORAD) score. As inflammatory markers, eosinophil cationic protein in serum and cytokine production by PBMCs were measured. RESULTS: After active treatment, 56% of the patients experienced improvement of the eczema, whereas only 15% believed their symptoms had improved after placebo (P =.001). The total SCORAD index, however, did not change significantly. The extent of the eczema decreased during active treatment from a mean of 18.2% to 13.7% (P =.02). The treatment response was more pronounced in allergic patients (at least one positive skin prick test response and elevated IgE levels), and in this group the SCORAD score decreased (P =.02 compared with nonallergic patients). During active treatment, serum eosinophil cationic protein levels decreased (P =.03). No significant changes in the production of the cytokines IL-2, IL-4, IL-10, or IFN-gamma were found. CONCLUSIONS: A combination of L rhamnosus 19070-2 and L reuteri DSM 122460 was beneficial in the management of AD. The effect was more pronounced in patients with a positive skin prick test response and increased IgE levels. PMID- 12589362 TI - Allergic potency of recombinant Fel d 1 is reduced by low concentrations of chlorine bleach. AB - BACKGROUND: Sodium hypochlorite (NaOCl), the primary component of household bleach, has been shown to alter the purified mouse allergen Mus m 1, such that antibody recognition, or immunogenicity, is lost. Results of initial experiments suggest that antibody recognition is lost at lower concentrations of NaOCl than those required to fragment Mus m 1. OBJECTIVE: We sought to determine whether NaOCl had similar effects on recombinant (r)Fel d 1 and whether the loss of antibody recognition correlated with the loss of biologic activity, as measured with a basophil histamine release assay. METHODS: Recombinant Fel d 1 was treated with increasing amounts of NaOCl, and the product of the reaction was analyzed by using SDS-PAGE, Western blotting, and ELISA. The biologic activity of NaOCl treated rFel d 1 was analyzed with a basophil histamine release assay. RESULTS: The protein fragmented at an NaOCl/rFel d 1 molar ratio of 7000, whereas cat specific IgG recognition was lost at a lower molar ratio of 560. Basophil histamine release assays were performed to determine the effect of NaOCl on the biologic activity of rFel d 1. An NaOCl/protein molar ratio of 70 caused a significant reduction in histamine release from basophils of subjects with cat allergy. A molar ratio of 140 further inhibited histamine release by rFel d 1, suggesting a dose-response relationship between NaOCl and loss of biologic activity. CONCLUSIONS: NaOCl modifies rFel d 1, resulting in loss of immunogenicity and attenuation of biologic activity, as measured by its ability to stimulate basophil histamine release. PMID- 12589363 TI - Current mite, cat, and dog allergen exposure, pet ownership, and sensitization to inhalant allergens in adults. AB - BACKGROUND: Simultaneous exposure to more than one allergen might modify the effect of individual allergens. OBJECTIVE: The purpose of this study was to investigate the effect of current exposures to mite, cat, and dog allergen and pet ownership on sensitization in adults. METHODS: Questionnaires, skin tests, and home visits (Der p 1, Fel d 1, and Can f 1, ELISA; mattresses, living room floors) were performed in 2502 adults. Allergen exposure was treated as a continuous variable and divided into quartiles. To investigate the interaction between allergens, quartiles for 3 allergens were added, creating arbitrary combined exposure categories. RESULTS: In the univariate analysis, mite sensitization was associated with Der p 1 in mattresses (odds ratio [OR], 1.10; 95% CI, 1.01 to 1.19; P =.03) and with Can f 1 in living room floors (OR, 1.08; 95% CI, 1.00 to 1.17; P =.05). In a multivariate regression analysis, Der p 1 in mattresses remained an independent associate of mite sensitization (OR, 1.12; 95% CI, 1.02 to 1.23; P =.03) and pollen sensitization (OR, 1.23; 95% CI, 1.11 to 1.36; P =.0001). The proportion of subjects sensitized to mite increased significantly with the increasing combined exposure categories (P <.0001). The highest prevalence of sensitization to cat and dog was in the medium combined exposure categories. Cat ownership was associated with a reduced prevalence of sensitization to cats (P =.002) and a reduced prevalence of sensitization to dog (P =.003) but had no effect on sensitization to mite and pollen. CONCLUSIONS: Sensitization to dust mites increased with the increasing combined exposure. Cat ownership was associated with a lower prevalence of sensitization to cat and dog but not to mite and grass pollen. PMID- 12589364 TI - House dust mite allergen in US beds: results from the First National Survey of Lead and Allergens in Housing. AB - BACKGROUND: Although exposure to house dust mite allergen is a major risk factor for allergic sensitization and asthma, nationwide estimates of dust mite allergen levels in US homes have not been reported. OBJECTIVE: The purpose of this study was to estimate the prevalence of dust mite allergen in beds of US homes and to identify predictors of dust mite allergen concentration. METHODS: Data were obtained from the first National Survey of Lead and Allergens in Housing, a cross sectional survey of 831 permanently occupied noninstitutional housing units that permitted resident children. Dust mite allergen concentration (Der f 1 plus Der p 1) was determined from a dust sample collected from a bed. The percentages of homes with concentrations at or greater than detection, 2.0 microg/g bed dust, and 10.0 microg/g bed dust were estimated. Independent predictors of allergen concentration were assessed with multivariable linear regression. RESULTS: The percentages of US homes with dust mite allergen concentrations at or greater than detection, 2.0 microg/g, and 10.0 microg/g were 84.2% (SE, 1.73), 46.2% (SE, 2.0), and 24.2% (SE, 2.1), respectively. Independent predictors of higher levels were older homes, non-West census regions, single-family homes, no resident children, lower household income, heating sources other than forced air, musty or mildew odor, and higher bedroom humidity. CONCLUSION: Most US homes have detectable levels of dust mite allergen in a bed. Levels previously associated with allergic sensitization and asthma are common in US bedrooms. Predictors can be used to identify conditions under which homes are more likely to have increased dust mite allergen levels. PMID- 12589365 TI - Carmine (E-120)-induced occupational asthma revisited. AB - BACKGROUND: Occupational asthma (OA) caused by carmine (E-120) has been reported. OBJECTIVE: We sought to evaluate the prevalence of sensitization and OA at a natural dye processing factory in which 2 workers had been given a diagnosis of carmine-induced OA 6 years previously. METHODS: The 24 current employees and one worker who had recently left work because of asthma completed a questionnaire and underwent skin testing (carmine, cochineal, carminic acid, curcuma, annato, and chlorophyll), carmine IgE dot-blot analysis, and methacholine inhalation testing. Workers exhibiting positive occupational skin test responses, work-related asthma, or bronchial hyperresponsiveness underwent specific inhalation challenge and serial peak expiratory flow rate recording. RESULTS: Positive skin test responses to carmine (41.7%), cochineal (29.2%), and carminic acid (4.2%) were observed. Carmine IgE dot-blot results were positive in 4 subjects. No difference in atopy or smoking was observed between occupationally sensitized and nonsensitized subjects. Among the 5 employees reporting work-related asthma, 2 had positive skin test responses, and 4 had bronchial hyperresponsiveness. Five subjects underwent specific inhalation challenges: 2 workers had early asthma responses to carmine and cochineal challenges, and the remaining subjects did not have suggestive peak expiratory flow recordings. The subject who had left his job was given a diagnosis of carmine-induced OA. CONCLUSION: The prevalence of sensitization and OA caused by carmine was 41.6% and 8.3%, respectively. When the 3 workers who had left their jobs were included, the cumulative incidence of sensitization and OA was 48.1% and 18.5%, resembling the healthy worker effect. Prevention programs to establish the permissible levels of airborne allergen should be implemented. PMID- 12589366 TI - Patients with anaphylaxis to pea can have peanut allergy caused by cross-reactive IgE to vicilin (Ara h 1). AB - BACKGROUND: Serologic cross-reactivity among legumes has been described; however, it is rarely clinically significant. In this study 3 patients with a history of anaphylaxis to pea are described who subsequently had symptoms after ingestion of peanut. OBJECTIVE: We investigated whether the peanut-related symptoms were due to cross-reactivity between pea and peanut proteins. METHODS: Peanut-related symptoms were documented according to case history or double-blind, placebo controlled food challenge results. Skin prick tests were performed, and specific IgE levels were determined for pea and peanut with the CAP system FEIA. IgE binding proteins in pea and peanut were identified by using immunoblot analysis. Cross-reactivity was studied by means of immunoblot and ELISA inhibition studies with whole extracts and purified allergens. RESULTS: Peanut-related symptoms consisted of oral symptoms in all patients, with additional urticaria and dyspnea or angioedema in 2 patients. All patients had a positive skin prick test response and an increased IgE level to pea and peanut. Immunoblotting revealed strong IgE binding to mainly vicilin in pea extract and exclusively to Ara h 1 in crude peanut extract. Immunoblot and ELISA inhibition studies with crude extracts, as well as purified proteins, showed that IgE binding to peanut could be inhibited by pea but not or only partially the other way around. CONCLUSION: Clinically relevant cross-reactivity between pea and peanut does occur. Vicilin homologues in pea and peanut (Ara h 1) are the molecular basis for this cross-reactivity. PMID- 12589367 TI - Sex differences in the pathogenesis of chronic urticaria. PMID- 12589368 TI - Structural similarity between the bee venom peptides and the immunodominant human myelin basic proteins: role for pathogenesis of acute disseminated encephalomyelitis. PMID- 12589369 TI - Treatment with omalizumab normalizes the number of myeloid dendritic cells during the grass pollen season. PMID- 12589370 TI - Gross pathology and histopathology of asthma. PMID- 12589372 TI - Efficacy of repetitive transcranial magnetic stimulation (rTMS) in the treatment of affective disorders. AB - Transcranial magnetic stimulation (TMS) is a relatively noninvasive technique to interfere with the function of small cortical areas through currents induced by alternating magnetic fields emanating from a handheld coil placed directly above the targeted area. This technique has clear effects on a whole range of measures of brain function and has become an important research tool in neuropsychiatry. More recently, TMS has been studied in psychiatry mainly to assess its putative therapeutic effects in treatment refractory major depression. Most studies indicate that both low-frequency TMS and higher (20 Hz) frequency repetitive TMS may have some antidepressant properties. However, definite therapeutic effects of clinical significance still remain to be demonstrated. PMID- 12589373 TI - A neurochemically distinct dorsal raphe-limbic circuit with a potential role in affective disorders. AB - The serotonergic system arising from the dorsal raphe nucleus (DR) has long been implicated in psychiatric disorders, and is considered one site of action of classical anxiolytic and antidepressant agents. Recent studies implicate the DR as a site of action of novel anxiolytic and antidepressant agents that target neuropeptide systems, such as corticotropin-releasing factor (CRF) and neurokinin 1 (NK1) antagonists. The present study identified unique characteristics of the dorsomedial DR that implicate this particular subregion as a key component of a circuit, which may be targeted by these diverse psychotherapeutic agents. First, it was observed that a cluster of CRF-containing cell bodies was present in the dorsomedial DR of colchicine-treated rats. Dual-labeling immunohistochemistry revealed that almost all CRF-containing neurons were serotonergic, implicating CRF as a cotransmitter with serotonin in this subpopulation of DR neurons. Moreover, dendrites laden with immunoreactivity for NK1 had a striking topographic distribution surrounding and extending into the dorsomedial subregion of the DR, suggesting that NK1 receptor ligands may selectively impact the dorsomedial DR. Finally, anterograde tract tracing from the dorsomedial DR combined with CRF immunohistochemistry revealed that CRF-containing axons from this subregion project to CRF-containing neurons of the central nucleus of the amygdala. Taken together, the present results reveal a circuit whereby NK1 receptor activation in the dorsomedial DR can impact on limbic sources of CRF that have been implicated in emotional responses. This circuit may be relevant for understanding the mechanism of action of novel psychotherapeutic agents that act through NK1 or CRF receptors. PMID- 12589374 TI - Nicotine induces glutamate release from thalamocortical terminals in prefrontal cortex. AB - It has been proposed that activation of nicotinic acetylcholine receptors (nAChRs) can activate the prefrontal cortex, enhancing attention and cognition. Nicotine can stimulate the release of several different neurotransmitters in many brain regions. In the present study, we found that stimulation of nAChRs by nicotine or the endogenous agonist, acetylcholine (ACh), induces a large spontaneous increase in glutamate release onto layer V pyramidal neurons of the prefrontal cortex. This release of glutamate, measured by spontaneous excitatory postsynaptic currents (sEPSCs) in the prefrontal cortical slice, depends on intact thalamocortical terminals. It can be suppressed by mu-opioids or eliminated by blocking action potentials. The increase in sEPSCs is sensitive to low concentrations of nicotine, suggesting the involvement of high-affinity (eg alpha(4)beta(2)) nAChRs. Recent work has shown alterations in prefrontal alpha(4)beta(2) nAChRs in autism and schizophrenia, two conditions that are distinguished by abnormal prefrontal cortical activation as well as difficulty in certain aspects of cognition and integrating social and emotional cues. We show that mice lacking the beta(2) nAChR subunit do not show increased sEPSCs with either nicotine or ACh, again implicating high-affinity nicotinic receptors. These findings give new insight into the mechanism by which nicotine affects excitatory neurotransmission to the output neurons of the cerebral cortex in a pathway that is critical for cognitive function and reward expectation. PMID- 12589375 TI - Sensitivity to sensorimotor gating-disruptive effects of apomorphine in two outbred parental rat strains and their F1 and N2 progeny. AB - Sensorimotor gating, measured by prepulse inhibition (PPI) of the startle reflex, is reduced in schizophrenia patients and in rats treated with dopamine agonists. Strain and substrain differences in the sensitivity to the PPI-disruptive effects of dopamine agonists may provide insight into the genetic basis for human population differences in sensorimotor gating. We have reported greater sensitivity to the PPI-disruptive effects of the D1/D2 agonist apomorphine in Harlan Sprague-Dawley (SDH) vs Wistar (WH) rats. In the present study, we assessed the inheritance pattern of this phenotypic difference. Sensitivity to the PPI-disruptive effects of apomorphine was compared across parental SDH and WH strains, offspring (F1) of an SDH x WH cross, and subsequent offspring (N2) of an SDH x F1 cross. Apomorphine sensitivity followed a gradient of SDH>N2>F1>WH. Parental SDH and WH strains exhibited comparable sensitivity to the PPI disruptive effects of phencyclidine. The nature of this gradient of APO sensitivity suggests relatively simple additive effects of multiple genes on the phenotype of PPI sensitivity. PMID- 12589376 TI - Reduction of hypothalamic vasopressinergic hyperdrive contributes to clinically relevant behavioral and neuroendocrine effects of chronic paroxetine treatment in a psychopathological rat model. AB - The neuroendocrine and behavioral effects of chronic paroxetine treatment were investigated in two rat lines selectively bred for high anxiety-related behavior (HAB) or low anxiety-related behavior (LAB) emotionality. In addition to a characteristic behavioral phenotype with markedly passive stress-coping strategies, HAB rats show a hypothalamic vasopressinergic hyperdrive that is causally related to hypothalamic-pituitary-adrenocortical dysregulation as demonstrated in the combined dexamethasone (DEX)/corticotropin-releasing hormone (CRH) test. A total of 8 weeks of chronic paroxetine treatment induced a more active coping strategy in the forced swim test in HAB rats only. In contrast, paroxetine-treated LAB rats did not change their swimming behavior. To investigate the neuroendocrine alterations linked to these behavioral changes, a combined DEX/CRH test was performed. In HAB rats, the paroxetine-induced behavioral changes towards more active coping strategies were accompanied by a normalization of the CRH-stimulated increase in corticotropin (ACTH) and corticosterone secretion. Concomitantly, the hypothalamic vasopressinergic hyperdrive was found to be reduced in HAB but not LAB rats, as indicated by a decrease in vasopressin mRNA expression, whereas vasopressin 1a receptor binding was unaffected. These findings provide the first evidence that the vasopressinergic system is likely to be critically involved in the behavioral and neuroendocrine effects of antidepressant drugs. This novel mechanism of action of paroxetine on vasopressin gene regulation renders vasopressinergic neuronal circuits a promising target for the development of more causal antidepressant treatment strategies. PMID- 12589377 TI - Imaging brain phospholipase A2 activation in awake rats in response to the 5 HT2A/2C agonist (+/-)2,5-dimethoxy-4-iodophenyl-2-aminopropane (DOI). AB - Incorporation coefficients k(*) of intravenously injected [(3)H]arachidonic acid from blood into brain reflect the release from phospholipids of arachidonic acid by receptor-initiated activation of phospholipase A(2) (PLA(2)). In unanesthetized adult rats, 2.5 mg/kg intraperitoneally (i.p.) (+/-)2,5-dimethoxy 4-iodophenyl-2-aminopropane (DOI), which is a 5-HT(2A/2C) receptor agonist, has been reported to produce the behavioral changes of what is known as the 5-HT(2) syndrome, but only a few small regional decrements in brain glucose metabolism. In this study, 2.5 mg/kg i.p. DOI, when administered to unanesthetized rats, produced widespread and significant increases, of the order of 60%, in k(*) for arachidonate, particularly in neocortical brain regions reported to have high densities of 5-HT(2A) receptors. The increases could be entirely blocked by chronic pretreatment with mianserin, a 5-HT(2) receptor antagonist. The results suggest that the 5-HT(2) syndrome involves widespread brain activation of PLA(2) via 5-HT(2A) receptors, leading to the release of the second messenger, arachidonic acid. Chronic mianserin, a 5-HT(2) antagonist, prevents this activation. PMID- 12589378 TI - Selective 5-HT1A antagonists WAY 100635 and NAD-299 attenuate the impairment of passive avoidance caused by scopolamine in the rat. AB - Systemic administration of the muscarinic-receptor antagonists atropine and scopolamine produces cognitive deficits in humans, nonhuman primates and rodents. In humans, these deficits resemble symptoms of dementia seen in Alzheimer's disease. The passive avoidance (PA) task has been one of the most frequently used animal models for studying cholinergic mechanisms in learning and memory. The present study examined the ability of two selective 5-HT(1A) receptor antagonists WAY 100635 and NAD-299 (robalzotan) and two acetylcholinesterase (AChE) inhibitors tacrine and donepezil to attenuate the impairment of PA retention caused by the nonselective muscarinic receptor antagonist scopolamine in the rat. Although demonstrating differences in their temporal kinetics, both WAY 100635 and NAD-299 attenuated the impairment of PA caused by scopolamine (0.3 mg/kg s.c.). Donepezil did not block the PA deficit caused by the 0.3 mg/kg dose of scopolamine, but it prevented the inhibitory effects of the 0.2 mg/kg dose of scopolamine. In contrast, tacrine was effective vs both the 0.2 and 0.3 mg/kg doses of scopolamine. These results indicate that (1). a functional 5-HT(1A) receptor antagonism can attenuate the anterograde amnesia produced by muscarinic receptor blockade, and (2). the AChE inhibitors tacrine and donepezil differ in their ability to modify muscarinic-receptor-mediated function in vivo. These results suggest that 5-HT(1A) receptor antagonists may have a potential in the treatment of cognitive symptoms in psychopathologies characterized by reduced ACh transmission such as Alzheimer's disease. PMID- 12589379 TI - Induction of metabolic hypofunction and neurochemical deficits after chronic intermittent exposure to phencyclidine: differential modulation by antipsychotic drugs. AB - Numerous human imaging studies have revealed an absolute or relative metabolic hypofunction within the prefrontal cortex, thalamus and temporal lobes of schizophrenic patients. The former deficit correlates with cognitive deficits and negative symptoms, whereas the latter correlates with positive symptomologies. There is also general consensus that schizophrenia is associated with decreased parvalbumin expression in the prefrontal cortex. Since the drug phencyclidine can induce a psychosis resembling schizophrenia in humans, we have examined whether repeated phencyclidine (PCP) treatment to rats could produce similar metabolic and neurochemical deficits to those occurring in schizophrenia and whether these deficits could be modulated by antipsychotic drugs. We demonstrate here that chronic intermittent exposure to PCP (2.58 mg kg(-1) i.p.) elicits a metabolic hypofunction, as demonstrated by reductions in the rates of glucose utilization, within the prefrontal cortex, reticular nucleus of thalamus and auditory system, key structures displaying similar changes in schizophrenia. Moreover, chronic PCP treatment according to this regime also decreases parvalbumin mRNA expression in the rat prefrontal cortex and reticular nucleus of the thalamus. Chronic coadministration of haloperidol (1 mg kg(-1) day(-1)) or clozapine (20 mg kg(-1) day(-1)) with PCP did not modulate PCP-induced reductions in metabolic activity in the rat prefrontal cortex, but reversed deficits in the structures of the auditory system. Clozapine, but not haloperidol, reversed PCP-induced decreases in parvalbumin expression in prefrontal cortex GABAergic interneurons, whereas both drugs reversed the deficits in the reticular nucleus of the thalamus. These data provide important new information, which strengthen the validity of chronic PCP as a useful animal model of schizophrenia, when administered according to this protocol. Furthermore, we propose that reversal of PCP-induced reductions in parvalbumin expression in the prefrontal cortex may be a potential marker of atypical antipsychotic activity in relation to amelioration of cognitive deficits and negative symptoms of schizophrenia. PMID- 12589380 TI - Inhibition of 5-HT neurotransmission increases clonidine protective effects on naloxone-induced conditioned place aversion in morphine-dependent rats. AB - Previous pharmacological studies have implicated serotonergic brain systems in opiate-withdrawal-precipitated conditioned place aversion. To assess this hypothesis, we tested the effects of either (i). a near-total 5,7 dihydroxytryptamine-induced lesion (90% depletion) or (ii). an acute serotonin (5 HT) inhibition induced by the specific stimulation of 5-HT1A autoreceptors (8 OHDPAT 5-100 microg/kg), on naloxone-induced conditioned place aversion in morphine-dependent rats. Morphine dependence was induced by the implantation of morphine slow-release pellets. The protective properties of clonidine (an alpha-2 adrenergic agonist classically given for opiate detoxification) were also tested after inhibition of 5-HT transmission. Serotonergic lesions in morphine-dependent rats failed to alter naloxone-induced conditioned place aversion but increased the sensitivity to the protective effects of clonidine. Acute neuropharmacological blockade of serotonin transmission also potentiated the clonidine effects on naloxone-induced conditioned place aversion. When combined with the 5-HT1A agonist 8-OHDPAT, clonidine was also found to be more potent. Further understanding of this serotonin/noradrenaline interaction might help devise new therapeutic treatments for the acute opiate withdrawal syndrome. PMID- 12589381 TI - Apomorphine enhances conditioned responses induced by aversive stimulation of the inferior colliculus. AB - Consistent evidence has shown that learning may be produced in paradigms using electrical stimulation of the inferior colliculus (IC) as unconditioned stimulus (UCS). Recent reports have also demonstrated that aversive stimulation of the IC, at the escape threshold, enhances dopamine (DA) release in the prefrontal cortex. The purpose of the present study was to determine whether dopaminergic mechanisms are involved in the Pavlovian conditioning and latent inhibition using IC stimulation as UCS and light as conditioned stimulus (CS). Rats were placed inside a shuttle box and subjected to a two-way avoidance paradigm. IC aversive electrical stimulation was used as UCS and shuttle box illumination as CS. The rats quickly learned to avoid or interrupt the IC stimulation. Apomorphine injections produced a dose-dependent increase in the number of avoidance responses. On the other hand, chlorpromazine administration promoted a dose dependent reduction of the avoidance responses. Previous injections of chlorpromazine inhibited the effects of apomorphine. Also, previous exposure to unreinforced light weakened the strength of the conditioning. Apomorphine blocked this latent inhibition effect, which was antagonized by previous injections of chlorpromazine. These findings bring evidence for the involvement of DA in the setting up of adaptive responses to aversive states generated at the IC level, which may underlie stressful situations present in anxiety. PMID- 12589382 TI - Enhanced morphine preference following prolonged abstinence: association with increased Fos expression in the extended amygdala. AB - We previously found that chronically morphine-pretreated, abstinent rats show stronger preferences for morphine-associated environments than placebo-pretreated rats. Here we show that this increased preference persisted for at least 5 weeks after withdrawal of chronic morphine. To determine brain regions involved in this behavior, we examined neural activation (as indexed by Fos-like proteins) induced by a morphine-conditioned place preference test. Placebo-pretreated (P) morphine conditioned rats showed significantly elevated Fos in the anterior cingulate cortex (Cg), nucleus accumbens core (Ac-C) and shell (Ac-S), ventral lateral and dorsal lateral bed nucleus of the stria terminialis (BNST-VL and -DL), and central and basolateral amygdala nuclei (ACE, ABL) when compared to nonconditioned P rats. Chronically morphine-pretreated (M) rats that exhibited enhanced morphine preference 5 weeks after morphine withdrawal showed significantly greater Fos in all the same areas except the BNST-DL relative to conditioned P or nonconditioned M rats. Place preference measures and Fos expression were positively correlated in the Cg and ABL, for conditioned P animals, and in the Cg, ABL and BNST-VL for conditioned M animals. These results indicate a relationship between place preference behavior and neural indices of activation in the forebrain in response to morphine-conditioned cues that may be chronically modulated by prior morphine exposure. PMID- 12589383 TI - Differential effects of haloperidol, risperidone, and clozapine exposure on cholinergic markers and spatial learning performance in rats. AB - Haloperidol (HAL), a potent typical antipsychotic, continues to be a frequently prescribed medication for behavioral disturbances associated particularly with schizophrenia despite well-documented adverse effects associated with its chronic use. Animal experiments have even indicated that HAL can damage cholinergic pathways and thus could be especially deleterious to those experiencing cognitive deficits. However, several recent clinical studies indicate that atypical antipsychotics may actually improve cognitive function in some patients, although this assertion requires further investigation. The purpose of this study was to compare the effects of prior chronic (45- or 90-day) oral exposure to HAL and the atypical antipsychotics risperidone (RISP) and clozapine (CLOZ) on cognitive performance and central cholinergic markers in rats. All analyses were done after 4 days of drug washout in order to minimize direct drug effects. Learning performance and choline acetyltransferase (ChAT) levels were assessed in a water maze task and with immunofluorescence staining, respectively. HAL significantly impaired learning performance after 90 but not after 45 days of treatment when compared to both vehicle controls and the atypical agents, while RISP slightly improved task performance. Both 45 and 90 days of previous HAL exposure reduced ChAT staining in several brain regions, including the cortex, caudate-putamen, and hippocampus. ChAT staining in the caudate-putamen and hippocampus was also decreased after 90 days of RISP exposure, raising the possibility of deleterious cognitive effects after exposure to this dosage for longer periods of time. The results suggest that antipsychotic drugs exert differential and temporally dependent effects on central cholinergic neurons and learning performance. PMID- 12589384 TI - Zaprinast, a phosphodiesterase 5 inhibitor, overcomes sexual dysfunction produced by fluoxetine, a selective serotonin reuptake inhibitor in hamsters. AB - A high incidence of sexual dysfunction among women is reported in the clinical literature. Little experimental investigation has been initiated on the ability of phosphodiesterase (PDE) inhibitors to overcome deficits in sexual functioning because of selective serotonin reuptake inhibitors (SSRIs). The effects of fluoxetine, an SSRI, and zaprinast, a PDE-5 inhibitor, on the lateral displacement response (used as a measure of sensitivity to reproductively relevant stimuli) of hamsters in behavioral estrus were investigated. In Experiment 1, hamsters that were maximally sensitive to reproductively relevant stimuli because they were at the peak of behavioral estrus were administered fluoxetine (10 mg/kg, i.p.); they had significantly decreased lateral displacement responses compared to vehicle-administered hamsters. In Experiment 2, hamsters that were relatively less sensitive to sexual stimuli because they were at the termination of behavioral estrus were administered zaprinast (3 mg/kg; i.p.); they had significantly enhanced lateral displacement responses compared to responses seen following vehicle administration. In Experiment 3, fluoxetine-induced deficits in the lateral displacement of hamsters at the peak of behavioral estrus were overcome by the coadministration of zaprinast. These data confirm previous findings that sexual dysfunction can be induced by SSRIs and extend the current knowledge to suggest that administration of a PDE-5 inhibitor can override SSRI-induced deficits in sexual functioning. PMID- 12589385 TI - Long-lasting changes in morphine-induced locomotor sensitization and tolerance in Long-Evans mother rats as a result of periodic postpartum separation from the litter: a novel model of increased vulnerability to drug abuse? AB - Daily postpartum separations from the litter produce enduring changes in anxiety and sensitivity to the antinociceptive effects of morphine in Long-Evans dams. We tested whether postpartum experience alters sensitivity to the effects of morphine on locomotor activity. Dams were tested 4-6 weeks after their pups were weaned, and had one of the following backgrounds: daily separation from the litter on postpartum days 2-14 for either 3 h (prolonged separation-LS) or 15 min (brief separation-BS), or no separation (nonhandled control-NH). After 2 consecutive days (B1-2) of baseline activity measurements, subjects were tested daily after s.c. injections of either morphine (10 mg/kg) or saline for 7 days and again on day 10. Beginning 5 days later, saline and 1.0-10 mg/kg of morphine were tested in all dams. On B1, LS and BS dams habituated slower than NH controls, yielding higher horizontal counts. LS dams failed to habituate across baseline days and were more active than other dams on B2. Sensitization, a progressive increase in horizontal activity, was more rapid and robust in LS and BS dams compared to NH animals. LS was the only group that developed tolerance to morphine-induced decreases in vertical activity. In LS dams with the history of morphine treatment, injection of saline resulted in higher horizontal activity and center time compared to saline-treated counterparts, indicative of conditioning. Among animals with a history of saline treatment, LS dams were more sensitive to morphine challenges than BS and NH dams. As a result of the robust and long-lasting increases in the ability of morphine to induce behavioral sensitization in litter-separated dams, periodic postpartum separation may represent a new animal model of increased vulnerability to substance abuse. PMID- 12589386 TI - Attenuation of cue-controlled cocaine-seeking by a selective D3 dopamine receptor antagonist SB-277011-A. AB - Conditioned stimuli (CS) previously paired with drugs of abuse can elicit cravings in humans, relapse to drug use, and can also reinforce drug-seeking behavior in both humans and animals, events that are believed to be subserved in part by activation of the mesolimbic dopamine system. Converging anatomical, pharmacological, and behavioral evidence implicates dopamine D(3) receptors in the mechanisms underlying cue-controlled behaviors. The purpose of the present study was therefore to investigate the effects on cocaine-seeking behavior of a novel D(3) receptor antagonist, SB-277011-A, which is 100-fold more selective for D(3) over D(2) dopamine receptors. We have established previously that second order schedules of reinforcement provide an animal model of cue-controlled drug seeking both prior to and after cocaine has been self-administered. SB-277011-A dose-dependently decreased cocaine-seeking maintained by a cocaine-associated conditioned reinforcer in both the first, drug-free interval and also following self-administration of cocaine. At higher doses, SB-277011-A also increased the latency to receive the first CS presentation and cocaine infusion, thereby decreasing the number of cocaine infusions self-administered under the second order schedule of reinforcement. SB-277011-A had no effect on cocaine intake under an FR-1 schedule of reinforcement, or on responding for sucrose under a second-order schedule of reinforcement, at any dose tested. These results therefore suggest that D(3) dopamine receptors may be critically involved in cue controlled drug-seeking behavior independently of any interaction with the reinforcing effects of cocaine itself, and may therefore provide a therapeutic target in the treatment of relapse to cocaine use induced by CSs. PMID- 12589387 TI - Acute changes in cerebrospinal fluid 5-HIAA following oral paroxetine challenge in healthy humans. AB - A number of studies have reported decreased human lumbar cerebrospinal fluid (CSF) concentrations of the major serotonin metabolite, 5-hydroxyindoleacetic acid (5-HIAA), following chronic administration of selective serotonin reuptake inhibitors (SSRIs). This decrease has been thought to be a consequence of elevated extracellular serotonin and to be mediated through terminal autoreceptor feedback inhibition of serotonin turnover. We wished to study the previously unexamined acute effects of SSRI administration on human CSF 5-HIAA. A serial lumbar puncture (LP) procedure was used to collect CSF samples before and after a single oral 40 mg dose of the SSRI paroxetine (PAR) or matching placebo in eight healthy adult humans in a randomized, double-blind fashion. CSF 5-HIAA concentrations did not change following placebo, but showed a statistically significant 27% mean increase 3 h following PAR. Our findings stand in contrast to the decreases reported for CSF 5-HIAA after chronic SSRI treatment in humans and the decreases seen in brain extracellular 5-HIAA after acute or chronic administration of SSRIs to animals. PMID- 12589388 TI - State markers of depression in sleep EEG: dependency on drug and gender in patients treated with tianeptine or paroxetine. AB - Tianeptine enhances while paroxetine inhibits serotonin reuptake into neurons; however, both show an antidepressive action. A subgroup of 38 depressed patients from a drug trial comparing the efficacy of tianeptine with that of paroxetine was studied with regard to their effects on sleep regulation, especially in relation to treatment response. We recorded sleep EEGs at day 7 and day 42 after the start of treatment with either compound, which allows measurement of changes due to the antidepressive medication in relation to the duration of treatment. Spectral analysis of the non-REM sleep EEG revealed a strong decline in the higher sigma frequency range (14-16 Hz) in male treatment responders independent of medication, whereas nonresponders did not show marked changes in this frequency range independent of gender. The patients receiving paroxetine showed less REM sleep and more intermittent wakefulness compared to the patients receiving tianeptine. REM density after 1 week of treatment was a predictor of treatment response in the whole sample. Psychopathological features with regard to the score in single items of the HAMD revealed predictive markers for response, some of which were opposite in the gender groups, especially those related to somatic anxiety. Changes in REM density were inversely correlated to the changes in HAMD in the paroxetine, but not the tianeptine, group. Our data suggest the importance of taking gender into account in the study of the biological effects of drugs. The study further points to the importance of the higher sigma frequency range in the sleep EEG of non-REM sleep and REM density as a marker of treatment response. PMID- 12589389 TI - A self-administered questionnaire to measure dependence on cigarettes: the cigarette dependence scale. AB - A valid measure of dependence on cigarettes is a useful tool for clinicians and researchers. The aim of this study was to develop a new, self-administered measure of cigarette dependence, and to assess its validity. The content of the instrument was generated in qualitative surveys. A long version (114 items) was tested on the internet in 3009 smokers. Subsamples provided retest data after 18 days (n=578), follow-up data after 45 days (n=990) and saliva cotinine (n=105). The study resulted in a 12-item scale labelled the Cigarette Dependence Scale (CDS-12), and in a 5-item version of this scale (CDS-5). Except for tolerance, CDS-12 covers the main components of DSM-IV and ICD-10 definitions of dependence: compulsion, withdrawal symptoms, loss of control, time allocation, neglect of other activities, and persistence despite harm. CDS-5 has similar measurement properties but less comprehensive content. Both scales had a high test-retest reliability (r>or=0.83), and a high internal consistency (Cronbach's alpha>or=0.84). CDS-12 scores were higher in daily smokers than in occasional smokers (+1.3SD units), and were associated with the strength of the urge to smoke during the last quit attempt (R(2)>or=0.25), and with saliva cotinine (R(2)>or=0.17). CDS-12 and CDS-5 scores decreased in daily smokers who switched to occasional smoking at 18-day retest. Dependence scores did not predict smoking abstinence at follow-up. In conclusion, CDS-12 and CDS-5 are reliable measures of cigarette dependence which fulfill several criteria of content validity and construct validity and are sensitive to change over time. PMID- 12589390 TI - Caffeine reversal of ethanol effects on the multiple sleep latency test, memory, and psychomotor performance. AB - Caffeine has been shown to reverse some of the performance-impairing effects of ethanol. However, it is not known whether this antagonistic effect of caffeine is mediated by a reduction in sleepiness. The present study assessed physiological alertness/sleepiness, memory, and psychomotor performance following the administration of placebo, ethanol, and caffeine+ethanol combinations. A total of 13 healthy individuals (21-35 years old) underwent four conditions presented in a Latin Square Design: placebo-placebo, ethanol (0.5 g/kg)-placebo, ethanol (0.5 g/kg)-caffeine 150 mg, and ethanol (0.5 g/kg)-caffeine 300-mg. The Multiple Sleep Latency Test (MSLT), psychomotor performance battery, memory test, and mood/sleepiness questionnaires were administered following each condition. The peak breadth ethanol concentration (BrEC) was 0.043+/-0.0197% and did not differ among the three caffeine treatments. As expected, ethanol reduced mean latency on the MSLT. The lowest caffeine dose reversed this effect and the highest dose increased mean latency (greater alertness) significantly beyond placebo levels. Ethanol also impaired psychomotor performance and memory. The 300-mg caffeine dose restored performance and memory measures to placebo levels. Although visual analog ratings of dizziness were increased by ethanol, they were not diminished by either caffeine dose. In conclusion, Low-dose caffeine prevented the sleepiness and performance impairment associated with a moderate dose of ethanol. Thus, caffeine, similar to other stimulants, can reverse the physiologically sedating effects of ethanol, although other negative effects remain. PMID- 12589391 TI - Steroid synthesis inhibition with ketoconazole and its effect upon the regulation of the hypothalamus-pituitary-adrenal system in healthy humans. AB - Steroid synthesis inhibitors are commonly used in the treatment of patients with Cushing's disease, but may also improve psychopathology in hypercortisolemic depressed patients. Since glucocorticoids exert a negative feedback at pituitary and supra-pituitary levels, the inhibition of steroid synthesis may lead to increased expression of corticotropin-releasing hormone (CRH) and arginine vasopressin (AVP). We studied the effect of treatment with 800 mg ketoconazole (3 weeks) upon the concentrations of basal plasma cortisol in the evening, corticosteroid-binding globulin (CBG), dehydroepiandrosterone-sulfate (DHEA-S), and ACTH as well as the concentrations of cortisol, CRH, and AVP in cerebrospinal fluid (CSF) at 8.30 h in 10 healthy, male volunteers. While we found cortisol plasma concentrations to be unchanged, we noted a significant increase in ACTH (post: 45.1+/-43.5; pre: 14.2+/-5.2 pmol/l; F(1,8)=9.78, p<0.02) and CBG concentrations (post: 38.8+/-4.3; pre: 31.9+/-4.2 microg/l), but DHEA-S plasma concentrations declined (post: 1.75+/-1.83; pre: 2.75+/-2.80 mg/l; F(1,8)=7.9, p<0.03). CRH concentrations in CSF were unchanged after treatment (post: 62.5+/ 15.9; pre: 63.7+/-13.9 pg/ml), while there was a trend for AVP concentrations to rise during treatment (post: 2.52+/-1.18; pre: 1.92+/-0.96 pg/ml; paired t=-1.9, p<0.1). Cortisol CSF concentrations declined in the elderly (pre: 52.5+/-23.2; post: 26.7+/-4.6 nmol/l), but not in the young subgroup (pre: 15.6+/-11.3; post: 27.7+/-9.4 nmol/l). We thus conclude that the treatment of healthy controls with steroid-synthesis inhibitors does not lead to a major increase in CRH secretion. PMID- 12589392 TI - Treatment of Tourette syndrome with delta-9-tetrahydrocannabinol (delta 9-THC): no influence on neuropsychological performance. AB - Previous studies provide evidence that marijuana (Cannabis sativa) and delta-9 tetrahydrocannabinol (Delta(9)-THC), the major psychoactive ingredient of marijuana, respectively, are effective in the treatment of tics and behavioral problems in Tourette syndrome (TS). It, therefore, has been speculated that the central cannabinoid receptor system might be involved in TS pathology. However, in healthy marijuana users there is an ongoing debate as to whether the use of cannabis causes acute and/or long-term cognitive deficits. In this randomized double-blind placebo-controlled study, we investigated the effect of a treatment with up to 10 mg Delta(9)-THC over a 6-week period on neuropsychological performance in 24 patients suffering from TS. During medication and immediately as well as 5-6 weeks after withdrawal of Delta(9)-THC treatment, no detrimental effect was seen on learning curve, interference, recall and recognition of word lists, immediate visual memory span, and divided attention. Measuring immediate verbal memory span, we even found a trend towards a significant improvement during and after treatment. Results from this study corroborate previous data suggesting that in patients suffering from TS, treatment with Delta(9)-THC causes neither acute nor long-term cognitive deficits. Larger and longer-duration controlled studies are recommended to provide more information on the adverse effect profile of THC in patients suffering from TS. PMID- 12589393 TI - Saccade and cognitive function in chronic kava users. AB - Kava is an extract from the Piper methysticum Forst. f. plant that has been consumed in the Pacific islands for millennia and more recently, among indigenous populations, in northern Australia and throughout the Western world as an herbal medicine. Through alterations on neuronal excitation, kava induces muscle relaxation, anasthesia, and has anxiolytic properties. There have been several isolated reports of psychotic syndromes, severe choreoathetosis and possible seizures following kava use. However, there is no conclusive evidence that kava interferes with normal cognitive processes. We tested a group of current, ex, and nonkava users among an indigenous population in northern Australia, using saccade and cognitive tests that have proven cross-cultural validity and are sensitive to subtle disruptions of the brain arising from substance abuse or neuropsychiatric illness. Despite collecting data from among the heaviest reported kava drinkers in the world, we found no impairment in cognitive or saccade function in individuals who were currently heavy kava users (and had been for up to 18 years), nor was there any impairment in individuals who had been heavy kava users in the past but had abstained for longer than 6 months. Current and ex-kava users showed a higher rate of kava dermopathy, lower body mass index, lowered blood lymphocytes and, in addition, current kava users showed elevated liver enzymes. While there has recently been increasing concern about potentially fatal liver damage attributed to kava use, we have found no evidence of brain dysfunction in heavy and long-term kava users. PMID- 12589394 TI - A BDNF coding variant is associated with the NEO personality inventory domain neuroticism, a risk factor for depression. PMID- 12589395 TI - Synergistic action of 5-HT2A antagonists and selective serotonin reuptake inhibitors in neuropsychiatric disorders. AB - Recently, the addition of drugs with prominent 5-HT(2) receptor antagonist properties (risperidone, olanzapine, mirtazapine, and mianserin) to selective serotonin reuptake inhibitors (SSRIs) has been shown to enhance therapeutic responses in patients with major depression and treatment-refractory obsessive compulsive disorder (OCD). These 5-HT(2) antagonists may also be effective in ameliorating some symptoms associated with autism and other pervasive developmental disorders (PDDs). At the doses used, these drugs would be expected to saturate 5-HT(2A) receptors. These findings suggest that the simultaneous blockade of 5-HT(2A) receptors and activation of an unknown constellation of other 5-HT receptors indirectly as a result of 5-HT uptake inhibition might have greater therapeutic efficacy than either action alone. Animal studies have suggested that activation of 5-HT(1A) and 5-HT(2C) receptors may counteract the effects of activating 5-HT(2A) receptors. Additional 5-HT receptors, such as the 5-HT(1B/1D/5/7) receptors, may similarly counteract the effects of 5-HT(2A) receptor activation. These clinical and preclinical observations suggest that the combination of highly selective 5-HT(2A) antagonists and SSRIs, as well as strategies to combine high-potency 5-HT(2A) receptor and 5-HT transporter blockade in a single compound, offer the potential for therapeutic advances in a number of neuropsychiatric disorders. PMID- 12589396 TI - Changes in human in vivo serotonin and dopamine transporter availabilities during chronic antidepressant administration. AB - Few studies have demonstrated in vivo alterations of human serotonin and dopamine transporters (SERTS and DATS) during antidepressant treatment. The current study measured these transporter availabilities with [(123)I]beta-CIT single photon emission computed tomography (SPECT) during administration of selective serotonin reuptake inhibitors (SSRIs) or a non-SSRI, bupropion. A total of 17 healthy human subjects were randomly assigned to two different treatment protocols: (1). citalopram (40 mg/day) followed by augmentation with bupropion (100 mg/day) or (2). bupropion (100-200 mg/day) for 16 days. Citalopram significantly inhibited [(123)I]beta-CIT binding to SERT in brainstem (51.4%) and diencephalon (39.4%) after 8 days of administration, which was similarly observed after 16 days. In contrast, citalopram significantly increased striatal DAT binding by 15-17% after 8 and 16 days of administration. Bupropion and its augmentation to citalopram did not have a significant effect on DAT or SERT. In 10 depressed patients who were treated with paroxetine (20 mg/day), a similar increase in DAT and inhibition of SERT were observed during 6 weeks treatment. The results demonstrated the inhibition of SERT by SSRI in human in vivo during the chronic treatment and, unexpectedly, an elevation of DAT. This apparent SSRI-induced modulation of the dopamine system may be associated with the side effects of these agents, including sexual dysfunction. PMID- 12589397 TI - Reversible disassembly of somatic nucleoli by the germ cell proteins FRGY2a and FRGY2b. AB - Egg cytoplasm has the capability to reprogramme differentiated somatic nuclei, as shown by nuclear transplantation in animal cloning. The nucleoli of donor nuclei are rapidly disassembled on injection into interphase eggs and are correctly reassembled when donor transcription initiates in the early embryos of frogs and mammals, recapitulating the physiological nucleolar dynamics of early embryogenesis. This is one of the most remarkable structural reorganizations of somatic nuclei in nuclear cloning. Despite the long history of nuclear cloning, almost nothing is known about the molecular mechanism of nucleolar disassembly in egg cytoplasm. Here we show that the Xenopus germ cell proteins FRGY2a and FRGY2b reversibly disassemble somatic nucleoli in egg cytoplasm, independently of continuing ribosomal RNA transcription. The carboxy-terminal domain of FRGY2a, which localizes to the nucleoli, is sufficient for nucleolar disassembly in transfected cells. Our results show that a single protein fragment can trigger reversible disassembly of the complex nucleolar structure. PMID- 12589398 TI - [Inhibitory effects of melatonin on the development of 17-beta-estradiol induced prolactinoma in relation to plasma prolactin and peroxidative lipid contents]. AB - In the present study, we have examined inhibitory effects of melatonin on the development of pituitary prolactin-producing tumors (prolactinoma) induced by 17 beta-estradiol (E(2)) in vivo. The prolactinomas were established by implanting E(2)-laden silastic capsules subcutaneously in Sprague-Dawley male rats weighing 80 approximately 100 g. Melatonin (0.05, 0.25, 0.50, 1.00, and 2.00 mg/0.1 ml/rat) was administrated subcutaneously at l8:00 h for 90 days, beginning from d 7 prior to tumor induction. Controls were given equal volumes of 4 percent; alcohol in 0.9 percent; saline. Our results showed: (1) In control group and groups given respectively 0.05, 0.25, 0.50, 1.00 and 2.00 mg melatonin, the weight of prolactinoma was 115.0+/-71.0, 85.2+/-41.0, 58.9+/-24.1, 72.7+/-23.6, 79.3+/-56.1, 74.5+/-46.8 mg respectively; the plasma prolactin (PRL) content was 493.46+/-33.3, 373.78+/-26.5, 125.13+/-13.3, 201.79+/-11.2, 418.88+/-41.3, 281.94+/-36.4 ng/ml respectively; the plasma peroxidative lipid content was 1.21+/-0.23, 0.89+/-0.32, 0.92+/-0.27, 0.64+/-0.24, 0.41+/-0.14 and 0.43+/-0.21 delta233/ml respectively. (2) The correlation coefficients between tumor weight and plasma PRL content, tumor weight and plasma peroxidative lipid content, and plasma PRL content and plasma peroxidative lipid content were 0.8738, 0.5550 and 0.2141 respectively. These results indicate: (1) The dosages of 0.25 (P<0.01) and 0.50 (P<0.05) mg, but not 0.05 (P>0.05), 1.00 (P>0.05) and 2.00 (P>0.05) mg, melatonin significantly inhibited the development of the E(2)-induced prolactinoma and the secretion of PRL in comparison with the matched control. (2) The levels of 0.05 approximately 2.00 (P<0.05 approximately 0.00l) mg melatonin showed a dose-dependent antioxidative action. (3) There are positive correlation between tumor weight and plasma PRL content (P<0.05), but no correlation between tumor weight and plasma peroxidative lipid content (P>0.05), and plasma PRL content and plasma peroxidative lipid content (P>0.05). Therefore, our experiments demonstrate that the inhibition of the development of E(2)-induced prolactinoma by adequate dosage of melatonin may be related to the inhibitory effects of MLT on the secretion of PRL, but not to the antioxidative action of MLT. PMID- 12589399 TI - [Effect of estrogen on dopamine release evoked by electric stimulation of central amygdaloid nucleus]. AB - In vivo fast cyclic voltammetry (FCV) was used to investigate dopamine (DA) release from central amygdaloid nucleus (CAN) of ovariectomized (OVX), estrogen treated and normal female rats. Radioimmunoassay was used to measure the serum concentration of estradiol (E(2)). DA release was significantly higher in OVX rats treated with estradiol benzoate (EB) as compared with controls. The DA release due to electric stimulation is dose-dependent on serum concentration of estradiol, suggesting that estrogen may play an important role in regulating DA release from CAN. PMID- 12589400 TI - [Hydrogen peroxide augments the injury effect of iron on the isolated rat heart and cardiomyocytes]. AB - By using Langendorff perfused rat heart and enzymatically isolated cardiomyocytes, we investigated the augmented injury effect of iron on the myocardium by hydrogen peroxide and the underlying mechanisms. Cell-permeable iron (Fe-HQ) decreased the contractile amplitude, velocity and end-diastolic cell length of the cardiomyocyte but increased the contents of lactate dehydrogenase (LDH) and creatine kinase (CK) in the coronary effluent and the myocardial malondialdehyde (MDA) while the left ventricular developed pressure (LVDP), +/ dp/dt(max), heart rate and coronary flow showed biphasic alterations. Hydrogen peroxide augmented the injury effect of iron accompanied by increases of coronary LDH, CK release and myocardial MDA content and decreases of LVDP, +/-dp/dt(max), and heart rate. Reduced glutathione could antagonize the injury effect of iron and hydrogen peroxide on the myocardium while dimethyl sulfoxide had no injury effect on the isolated heart. It is suggested that the functional injury of sulfhydryl group containing proteins may be involved in the augmentation of myocardial injury due to the increase of intracellular iron by hydrogen peroxide, but hydroxyl radicals may not. PMID- 12589401 TI - [Therapeutic angiogenesis with the use of vascular endothelial growth factor 165 gene in the myocardium of miniature swine]. AB - The purpose of the present study was to examine whether adenovirus-mediated vascular endothelial growth factor l65 (VEGFl65) can enhance collateral vessel formation of coronary artery and improve regional myocardial perfusion and function. Using a miniature swine model of chronic myocardial ischemia, the replication-deficient recombinant adenovirus vector containing complementary deoxyribonucleic acid (cDNA) for human VEGFl65 (Ad-VEGFl65) or for beta galactosidase (Ad-Gal) was administered directly into the ischemic myocardium in the left circumflex (LCX) distribution. Myocardial perfusion and function were assessed by electrocardiogram-gated single photon emission computed tomography (SPECT) imaging and collateral vessel development of coronary artery was assessed by ex vivo coronary angiography (CAG). Four weeks after Ad-VEGF165 administration, SPECT imaging demonstrated a significant reduction in ischemic area (P<0.01) and ischemic severity (P<0.01), and a substantial improvement in left ventricular ejection fraction (P<0.01) and regional wall motion in the LCX distribution (P<0.05), as compared with that of control animals and that before administration of Ad-VEGFl65. Collateral vessel development with Rentrop Grading was also significantly greater in Ad-VEGF165 animals than in the Ad-Gal control animals (P<0.05). It's concluded that Ad-VEGFl65 can induce collateral vessel development in ischemic myocardium and result in significant improvement in myocardial perfusion and function. PMID- 12589402 TI - [Characteristics of alpha2-adrenoceptor mediated contractile response in isolated aortae of rats]. AB - In order to investigate the characteristics of vascular alpha(2)-adrenoceptor (alpha(2)-AR) and its relation to alpha(1)-AR, isolated Wistar rat (10 weeks) aortae were used as a model for testing contractile function in vivo. It was found that both alpha(1) and alpha(2)-AR (mainly alpha(1)-AR) mediated the contractile response. While alpha(1)-AR enhanced the contractile response mediated by alpha(2)-AR, alpha(2)-AR had no effect on that mediated by alpha(1) AR. When alpha(1)-AR was irreversibly blocked and the activity of alpha(2)-AR remained intact, the contractile response mediated by alpha(2)-AR was no longer observed. The contractile response only reappeared in the presence of KCl of the threshold level, and the extent of the maximum contraction decreased, compared with control (with alpha(1)-AR being intact). The results show that there exists a functional alpha(2)-AR in rat aorta, however, the contractile effect of which depends on the stimulation of alpha(1)-AR. PMID- 12589403 TI - [Relationship between heat stress suppression of neuroapoptosis and activation of nuclear kappa B in primary cultured rat cerebellar granule cells]. AB - It has been well demonstrated that heat stress response (HSR) plays a crucial role in protecting cells from injury induced by various pathological stimuli. However, the protective mechanism of HSR is only poorly understood. The object of this article was to further investigate the relationship between the protective role of heat stress response and activation of NF-kappa B in primary cultured rat cerebellar granule cells. Heat stress was induced by hyperthermia (43+/-0.5 degrees centigrade), and DNA binding activity of NF-kappa B was determined with electrophoretic mobility shift assay (EMSA). Neuroapoptosis was measured by Hoechst 33258, agarose gel electrophoresis and flow cytometry (FCM) analysis. The results showed that the neurons treated with low potassium for l6 h could induce neuroapoptosis and promote the activity of nuclear kappa B. Heat stress treatment for 30, 60 and 90 min could suppress neuroapoptosis and the activity of nuclear kappa B induced by low potassium in a time-dependent manner. Activation of NF kappa B using 100 nmol/L phorbol 12-myristate l3-acetate (PMA) could promote antiapoptotic action of heat stress response. In contrast, when NF-kappa B activation was inhibited by 10 micromol/L pyrrolidine dithiocarbamate derivatives (PDTC), heat stress did not provide protection against cell apoptosis induced by low potassium. The results suggest that the neuroprotection of heat stress has no relation to the suppression of NF-kappa B activity, and activation of NF-kappa B may promote antiapoptotic action of heat stress. PMID- 12589404 TI - Suppression of large conductance Ca2+-activated K+ channels by amyloid beta protein fragment 31-35 in membrane patches excised from hippocampal neurons. AB - To clarify the shortest essential active sequence in amyloid beta-protein (AbetaP) responsible for affecting neuronal electrophysiological properties, the effects of fragments 31-35 and 25-35 of AbetaP on the large conductance Ca(2+) activated potassium (BK) channels were investigated in the "inside-out" membrane patches excised from hippocampal neurons of rats. After application of AbetaP 3l 35 (5 micromol/L, n=10), the mean P(o) and open frequency of BK channels decreased by 85.8+/-l3.5 percent;percent; (P<0.01) and 72.1+/-22.8 percent; (P<0.01), respectively, and the mean open time decreased by 41.l+/-l8.5 percent; (P<0.0l), while the mean current amplitude was not significantly affected (P>0.05). Application of AbetaP 25-35 (5 micromol/L) also induced a decrease of 85.5+/-22.l percent; (P<0.0l) in mean P(o) and of 5l.4+/-18.3 percent; (P<0.05) in mean open time within l~3 min after application. These results suggest that the functional alteration in BK channels elicited by AbetaP fragments may play an important role in the mechanisms underlying AbetaP neurotoxicity, and AbetaP 31 35 may be the shortest active sequence in AbetaP responsible for affecting the electrophysiological properties of neurons. PMID- 12589405 TI - [Inhibition of ATP-activated current by met-Enk in isolated DRG neurons of the rat]. AB - The present study aimed to explore modulation of ATP-activated currents (I(ATP)) by met-Enk in rat DRG neurons. I(ATP) was recorded using the whole-cell patch clamp technique. The majority of the neurons examined responded to ATP (90.0 percent;, 45/50) with inward currents. In the 45 ATP sensitive neurons three kinds of responses to application of met-Enk were distinguished: (l) inward currents (29/45), (2) outward currents (9/45), and (3) no effect (7/45). Pretreatment with met-Enk (10(-9)~10(-5) mol/L) suppressed I(ATP) (10(-4) mol/L) in 29 neurons responding to met-Enk with inward currents. The inhibition by met Enk of I(ATP) could be blocked by naloxone (10(-7) mol/L) in a concentration dependent manner. Met-Enk of 10(-9), 10(-8), 10(-7), 10(-6) and 10(-5) mol/L suppressed I(ATP) by l3.2+/-5.4 percent; (n=5); 39.2+/-8.6 percent; (n=8), 54.l+/ 8.6 percent; (n=8),43.3+/-7.9 percent; (n=7) and 43.l+/-7.9 percent; (n=7) (mean+/-MSE), respectively. A comparison of concentration - response relations of ATP with and without preapplication of met-Enk indicated that after pretreatment with met-Enk (10(-7) mol/L) the curve shifted downward markedly with a decrease of 25 percent; of the maximum value of I(ATP) and unchanged K(d) value. The suppression of I(ATP) by met-Enk was reversed as evidenced by intracellular dialysis of H-9 by using the repatch technique. Taken together, it is suggested that the inhibition by met-Enk of I(ATP) is caused by activation of opiate receptor, which eventually results in phosphorylation of ATP receptor, mediated by modulation of G protein coupling and intracellular signal transduction. PMID- 12589406 TI - [Modulatory effect of orphanin FQ on the IL-1beta transcripts in hippocampus]. AB - Using in situ hybridization and immuno-fluorescent double labeling method, we investigated the function of orphanin FQ on interlenkin-lbeta (IL-lbeta) transcripts in hippocampus. Intracerebroventricularly (icv) administrated IL lbeta antibody reduced IL-l and TNF-alpha secreted from peritoneal macrophage. The high level of IL-lbeta transcript in hippocampus elicited by traumatic stress was blocked by icv injection of orphanin FQ (0.55 nmol), which was reversed by orphanin FQ receptor antagonist ([phe(1)Psi(CH(2)-NH)Gly(2)] nociceptin-(1-l3) NH(2)). Opioid receptor-like receptor transcripts were found in neuron, astrocyte and microglia. Based on the results, we conclude that orphanin FQ functions as a neuroimmune modulator, and provokes immune response via mediation of IL-lbeta derived from neuron, astrocyte and microglia in hippocampus. PMID- 12589407 TI - [Effect of nucleus raphe obscurus on myoelectric activity of sphincter of Oddi in rabbits]. AB - The aim of the present study was to investigate the role of nucleus raphe obscurus (NRO) in regulating the motility of sphincter of Oddi (SO). After fasting about l8~24 h, the rabbits were anesthetized with urethane (1.0 g/kg), and the myoelectric signals of SO were induced by a pair of copper electrodes inserted into the subsera. The results of microinjection of various drugs into NRO are as follows. After glutamate (340 mmol/L, 0.1 microl) was injected, activity of SO was excited. With microinjection of GABA (1 mol/L,0.1 microl), the spike burst of SO was inhibited. Following microinjection of ketamine (180 mmol/L, 0.1 microl), a kind of N-methyl-D-aspartate (NMDA) receptor antagonist, SO motility was inhibited and the effect of glutamate was abolished. Injection of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (2 mmol/L, 0.1 microl), a non-NMDA receptor antagonist, also excited the myoelectric activity of SO, but did not inhibit the effect of glutamate. The effect of glutamate was abolished by intravenous injection of atropine (0.2 mg/kg) or bilateral vagotomy, but not by injection of phentolamine (1.5 mg/kg) or propronalol (1.5 mg/kg), or by transection of the spinal cord. The above results indicate that NRO mediation of SO activity is due to the effect of glutamate on the NMDA receptors in the nucleus, the output of which is sent through vagal nerve and peripheral M cholinergic receptor to exert excitation of gallbladder motility. PMID- 12589408 TI - [Changes in SR Ca2+-ATPase activity, Ca2+ release and uptake kinetics of diaphragm muscle after different-frequency chronic electrical stimulation of diaphragmatic nerve]. AB - To study adaptation of rabbit diaphragm muscle after different-frequency chronic electrical stimulation, Ca(2+)-ATPase activity and Ca(2+) release-uptake kinetics of sarcoplasmic reticulum (SR) were respectively measured by detecting inorganic phosphorus ion and Furo-2 fluorescence. SR Ca(2+)-ATPase activity of the low frequency stimulation group was significantly lower than that of the control group (P<0.0l), but it was significantly higher in the high-frequency stimulation group against control (P<0.0l). The kinetics of Ca(2+) release and Ca(2+) uptake was significantly lower in low-frequency group than that of the control (P<0.0l), but the kinetics of Ca(2+) release and Ca(2+) uptake was significantly higher than that of the control (P<0.01). It is thought that different-frequency electrical stimulation induced different adaptative changes in SR Ca(2+)-ATPase activity, and Ca(2+) release and uptake kinetics of rabbit diaphragm muscle. PMID- 12589409 TI - Characteristic behavioral seizures and abnormal signal asymmetry of magnetic resonance imaging in an electrogenic rat model of chronic epilepsy. AB - Chronic tetani (60 Hz, 2 s, 0.4~0.6 mA) were administered to the dorsal hippocampus (DHPC) or the medial temporal lobe neocortex (MTNC) of rats, to study the role of the entorhinal cortex (EC)-hippocampal loop in temporal lobe epileptogenesis. This was repeated once a day for 7 or 10 days. Magnification of hyper-intensity was induced by tetanization of the HPC or the MTNC, as detected by contralateral T(2) weighed magnetic resonance imaging (T(2)-WI). The effects were associated with an enlarged volume of the lateral ventricle (LV), which was verified histologically. T(2)-WI hper-intensities, contralateral to the tetanized hemispheres, were observed with high frequency primary wet dog shakes (WEDS) in the DHPC-stimulated rats and with low frequency WEDS in the MTNC-stimulated rats. It seems likely that the same neural mechanisms are shared by chronic tetanization of the right HPC and the righ MTNC, involving the closed EC-HPC loop. Poor correlation between contralateral T(2)-WI hper-intensities and light primary behavioral seizures in the MTNC-stimulated rats might be attributed to a controlled information flow into or out of this loop because of potential EC gating. In addition, asymmetric T(2)-WI hyper-intensities in the LV area reflected a hemispheric dependence, contralateral to the electrogenic focus in our model of rat epilepsy. PMID- 12589410 TI - [Roles of Galphaq/11 mediated- and platelet-derived growth factor mediated-signal transduction pathways in rat aorta restenosis]. AB - To observe the roles of Galphaq/11 mediated- and platelet-derived growth factor (PDGF) mediated-signal transduction pathways in proliferation and migration of vascular smooth muscle cells (VSMC) after arterial injury, a vascular cell proliferation model was established by balloon injury in rat aorta and the morphologic changes in injured vascular walls after the injury were studied. Activities of angiotensin converting enzyme (ACE) and aorta phospholipase C (PLC) were tested, and levels of platelet-derived growth factor receptor-beta (PDGFR beta) and Galphaq/11 protein were measured by Western blot analysis. At l day after operation, injured aortic segments showed denudation in endothelial cells. Medial VSMC proliferation and intimal hyperplasia were not observed. As compared with the sham group, ACE activities were increased by 382.7 percent; (P<0.0l), but the expression of PDGFR-beta and PLC activities did not show significant changes (P>0.05). In addition, the level of Galphaq/11 protein was decreased by 20.0 percent; (P<0. 05). At l4 days after operation, sections of injured aorta showed marked intimal thickening with large numbers of VMSCs proliferating throughout intima and media. In comparison with the sham group, ACE activity, PLC activity and the level of PDGFR-beta were increased by 420.2 percent; (P<0.01), 186.2 percent; (P<0.05) and 85.0 percent; (P<0.05), respectively. While the level of Galphaq/11 protein was decreased by 33.1 percent; (P<0.01). The above data suggest that the PDGF-mediated signal transduction pathway plays an important role in VSMC proliferation. PMID- 12589411 TI - Possible role of dentate gyrus in greneration of rat temporal lobe epilepsy induced by electrical stimulation. AB - Possible role of the dentate gyrus (DG) and the hippocampus (HPC) in temporal lobe epileptogensis was investigated in an electrogenic model of rat epilepsy. Chronic tetani (60 Hz, 0.4-0.6 mA. 2 s) were administered once daily for 7 days to the right dorsal hippocampus (DHPC) or the right DG. Animal behavior was observed and depth electro-graphic seizures and T(2)-weighted magnetic resonance images (T(2)-WI) were measured. Results indicated that the frequency of primary wet dog shakes (WEDS) in the DG-stimulated rats was much lower than that in the DHPC-stimulated rats (P<0.05). The mean maximal wave-amplitude in DG electrographs was also much lower than that in HPC electrographs (P<0.05). The oscillations proportion of DHPC electrographs increased after DHPC-tetanization (from 2/9 up to 7/9 rats). T(2)-WI hyperintensity in the lateral ventricle area was detected only in the DHPC-tetanized rats, not in the DG-tetanized rats (P<0.05). These results suggest that the DG acts as a filtering site in the entorhinal cortex-HPC neuronal circuitry and its dysfunction causes damage to the HPC and the generation of temporal lobe epilepsy. PMID- 12589412 TI - [Protective effect of Angelica on ECV(304) from injury induced by hyperlipidemic serum in vitro]. AB - The aim of this article was to examine the protective effect of Chinese traditional medicine Angelica on human umbilical vein endothelial cells (HUVECs, ECV(304)) from injury induced by hyperlipidemic serum (HLS) and to study the underlying mechanisms. Microstructures of HUVECs were observed by a scanning electron microscope. Spectrophotometer and immunocytochemical methods were used to detect the content of NO in the suspension and expression of ICAM-1, TGFbeta(1), bFGF on the cell surface, respectively. After being incubated with HLS for 24 hours, HUVECs exhibited pronounced morphological changes, such as disappearance of microvilli on the endothelial cell (EC) surface, rupture of cell membranes, etc. Expression of ICAM-l and bFGF in ECs was significantly increased, while expression of TGFbeta(1) and the release of NO from ECs were significantly decreased. All these effects of HLS on ECs can be reversed by Angelica significantly. The above effects of Angelica may be related to its anti atherosclerotic action. PMID- 12589413 TI - [Is the clinical application of stem cells within reach?]. PMID- 12589414 TI - [Objective diagnosis of central auditory processing and perception disorder]. PMID- 12589415 TI - [Teleteaching in otorhinolaryngology. Part 2: the European database "medicstream"]. AB - Searching the medical literature is mainly characterized by the use of databases. The time for reviewing and printing a paper causes a delay between the first presentation of scientific results in a lecture and the appearance of the appropriate journal publication. Moreover, the possibilities for the integration of multimedia data are limited. Because of the development of new compression algorithms for audiovisual digitization software, the near simultaneous transmission of scientific presentations, both visually and aurally, has been possible since 1999. The section telemedicine of the interdisciplinary study group on image guided surgical navigation from the University of Leipzig has developed "medicstream" as a unique, audiovisual, scientific internetdatabase. It allows the documentation of presentations at congresses and courses, including the discussion,over a freely accessible, widely available homepage, using a home developed streaming technology. All presentations are examined by an authorized editorial committee consisting of representatives with different medical specializations for content before the admission to the data base. A total of 392 presentations from seven scientific meetings can be selected by an integrated search machine and viewed with Windows Media Player or Real Player using a conventional or faster internet connection to the homepage www.medicstream.de. The quality of the audiovisual transmission depends on the receiver-lateral data transmission rate,whereby the minimum variant of 56 kB/s is characterized by good detectability of the contents. In an analysis period of 242 days,we registered 44,199 accesses and 4,488 attendances. The telemedicine data base "medicstream" can optimize quality and extend medical education by live-streaming as well as by archiving scientific presentations as audiovisual data. PMID- 12589416 TI - [Deafness in the cultural-historical context. Part 2]. PMID- 12589417 TI - [Argon plasma coagulation and topically applied estriol. Long-term results in the treatment of hereditary hemorrhagic telangiectasia of the nasal mucosa]. AB - BACKGROUND AND OBJECTIVE: Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant fibrovascular dysplasia with the main symptom of recurrent epistaxis. At present, only limited data are available on long-term results in the treatment of epistaxis. A part from the surgical treatment of the bleeding telangiectasia with argon plasma, an additional postoperative long-term treatment with estriol nose-ointment was performed and the results were analysed. PATIENTS AND METHODS: 69 patients with HHT were treated with APC and estriol nose ointment. 43 patients could be examined over a follow-up period of more than 20 months. The bleeding frequency and intensity as well as the patient's satisfaction with the treatment result were evaluated and the serum estriol level determined. RESULTS: After more than 20 months, the bleeding frequency and intensity were reduced in 95% of the patients compared to their preoperative findings. Under the influence of estriol, former berry-like telangiectasias became flatter and more even. Neither systemic side effects under topically applied estriol nor a relevant increase of serum estriol levels were observed. CONCLUSION: The combined treatment regimen with APC and topically applied estriol provides a new treatment concept of epistaxis in HHT. PMID- 12589418 TI - [Examination of binaural signal processing in normally hearing subjects using electrophysiological and psychoacoustical measurements]. AB - BACKGROUND AND OBJECTIVE: At present, only a small number of validated, clinically usable methods for the assessment of binaural hearing capabilities exist. A proposed electrophysiological measure is the registration of the brainstem-based binaural difference potentials (BDP). PATIENTS/METHODS: The BDP is calculated as the difference between the binaurally evoked registration and the sum of the two monaural registrations. Detection and stability of the BDP were examined in 24 normally hearing adults within the framework of conventional registration of auditory brainstem responses. Furthermore, the influence of interaural time differences (ITD) on the BDP was determined. In addition, lateralization of the subjects was assessed using a psychoacoustical method. RESULTS: The components of the BDP could be detected in almost all of the subjects. Moreover, they showed sufficient test-retest reliability. The impact of ITD,which causes lateralization of the stimulus,was clearly detectable for the latencies and the amplitudes of the BDP. CONCLUSIONS: Binaural difference potentials, which are easily and reliably detectable reveal a relationship to the outcome of psychoacoustical assessment of lateralization and have the potential to provide a measure for binaural hearing capacity. PMID- 12589419 TI - [Preservation of mucosal specimens before processing in the alkaline single cell microgel electrophoresis assay]. AB - BACKGROUND: Human mucosal biopsies are established in ecogenotoxicological studies, but up until now they have demanded immediate processing after harvesting. We report our experience with the preservation of specimens either for 24 h at 4 degrees C or for longer periods at -80 degrees C and compare the results with fresh specimens using the alkaline single cell microgel electrophoresis assay. PATIENTS AND METHODS: Nasal mucosa was harvested from ten patients, transferred to the laboratory and divided into groups for immediate processing,24 h preservation at 4 degrees C and cryopreservation at -80 degrees C. Alkaline single cell microgel electrophoresis assays were performed after separating the specimens into single cells and after exposure to benzo[a]pyrene,benzo[a]pyrene-diolepoxide, N-nitrosodiethylamine, or sodium dichromate. The trypan blue exclusion test was used to assess cytotoxicity. RESULTS: Despite of the fact that cell viability remained stable, after cryopreservation DNA-migration increased significantly for the negative control and benzo[a]pyrene. Although an increase was also seen for sodium dichromate, this was not significant. For benzo[a]pyrene-diolepoxide, N-nitrosodiethylamine and N-methyl-N'-nitro-N-nitrosoguanidine there were no significant changes in DNA migration. After 24 h in cell medium at 4 degrees C,DNA-migration did not rise compared to the samples which were immediately processed. CONCLUSIONS: Preservation of mucosal specimens at 4 degrees C for 24 h may be legitimate in order to facilitate laboratory practice. However, cryopreservation should not be applied because it leads to higher rates of DNA migration in some tested substances in the alkaline single cell microgel electrophoresis assay. PMID- 12589420 TI - [An intraosseous hemangioma in the perpendicular lamina of the ethmoid bone]. AB - The incidence of intraosseous hemangiomas is very low. Most of them occur in adult females. The mandible,the zygoma,the maxilla, and the frontal and nasal bones are the areas of most frequent localization in the facial region. The intraosseous hemangioma is a benign, slowly growing,bony hard tumor causing facial deformity. It is diagnosed histologically. We report a case of an 18-year old male with an intraosseous cavernous hemangioma located in the perpendicular lamina of the ethmoid bone. Intraosseous hemangioma often has characteristic signs on a CT scan, with either coarsened trabeculae lying adjacent to the vascular channels or multifocal lytic areas creating a honeycomb pattern. Because of the age of the patient we considered a midline granuloma and a chondrosarcoma. The therapy of choice is surgical excision; radiation is done in exceptional cases only. PMID- 12589421 TI - [Management of unilateral traumatic bulb luxation with disruption of the optic nerve]. AB - Luxation of the eyeball, which means that the eye protrudes between the eyelids,may occur spontaneously or as a rare sequel to major trauma to the orbital wall. It can be accompanied by a disruption of the optic nerve, which is called optic nerve avulsion. In most cases primary enucleation is performed as therapeutic treatment. In our case of a 1 1/2-year-old child bulb luxation with disruption of the optic nerve followed a relatively slight trauma,presumably a fall onto a toy. An irreversible amaurosis was diagnosed. To guarantee symmetrical growth of the orbita and face, the eyeball was replaced into the orbit under microscopic and endoscopic vision via a transnasal and transethmoidal approach.Postoperative examinations showed an exotropy with unrestricted motility of the globe and amaurosis. Five years after the trauma we found symmetrical facial development with uniform orbital cavities. The clinical features and management of globe luxation are discussed, importance of a rapid reposition of the eyeball in contrast to primary enucleation for functional, cosmetic and psychological reasons is emphasized. PMID- 12589422 TI - [Normal nasalance for the German language. Nasometric values for clinical use in patients with cleft lip and palate]. AB - OBJECTIVE: Rhinophonia is often present in patients with cleft lip and palate (CLP) many years after palatoplasty. It may be caused either by organic deficits, functional disorders, or both. NasalView is known as an instrument for the objective assessment of nasalance in cases of velopharyngeal insufficiency or incompetence.However,normative values for the German language are not yet available. In order to overcome this problem, this study evaluated such norms using NasalView. METHODS: A total of 50 individuals with normal speech development were examined with NasalView. The median age was 14 years (range 11 20 years). The tone material used comprised the vowels /a:/, /e:/, /i:/, /o:/, /u:/, the sentences S(1): "Die Schokolade ist sehr lecker."("This chocolate is very tasty.") and S(2): "Nenne meine Mamma Mimmi."("Call my mummy Mimmi."), and the text passages of LT(1): "Nordwind und Sonne"("Northwind and sun"), LT(2): "Kindergeburtstag"("A child's birthday party") and LT(3): "Der grosse Gesang"("A famous song"). RESULTS: The mean nasalance for the vowels was 35.9% (+/-8.4), for S(1) (containing no nasal consonants) 24.9% (+/-5.3), and for S(2) 69.6% (+/-5.5) (with many nasal sounds). The results for the text passages were 42.1% (+/-4.2) for LT(1), 36.9% (+/-4.3) for LT(2) and 38.2% (+/-4.4) for LT(3). CONCLUSION: The norms of nasalance for the German language presented here may be useful in measuring the long-term outcome of cleft lip and palate. PMID- 12589423 TI - [Reduction in fees according to GOA section 6a also applies to external physicians]. PMID- 12589424 TI - [Systematic investigation technique in videolaryngoscopy and stroboscopy]. PMID- 12589425 TI - [Haemostatic disorders in ENT patients. Part 1]. PMID- 12589426 TI - Gene rearrangements in the glucocerebrosidase-metaxin region giving rise to disease-causing mutations and polymorphisms. Analysis of 25 Rec NciI alleles in Gaucher disease patients. AB - The glucocerebrosidase and metaxin genes lie in a gene-rich region that also includes two corresponding pseudogenes. This gives rise to recombinant alleles. We analysed two groups of patients from Argentina and Spain: 25 bearing the Rec NciI allele and 36 carrying L444P. The mutational mechanism is described and the crossover site precisely defined. Most of the Rec NciI alleles were generated by gene conversion. Rearranged alleles involving the metaxin gene were also identified. The high frequency of Rec NciI alleles associated with a polymorphic rearrangement at the metaxin level is probably due to a founder effect. PMID- 12589427 TI - Notch4, a non-HLA gene in the MHC is strongly associated with the most severe form of alopecia areata. AB - Alopecia areata (AA) is a disorder primarily affecting the hair and nails in which associated autoimmune or atopic disease is common. Genetically, it is a complex trait with evidence of a role for genes of the major histocompatibility complex (MHC), the interleukin-1 cluster and chromosome 21 in the pathogenesis. The strongest association is with HLA class II alleles, although whether this indicates a direct contribution to the pathogenesis or results merely from linkage disequilibrium with nearby disease genes is unknown. Notch4 is a recently defined gene in the HLA class III region. Notch signalling is a direct determinant of keratinocyte growth arrest and entry into differentiation. A possible role for Notch in hair growth has been indicated by transgenic mouse findings that activation of the Notch pathway in the hair cortex leads to aberrant differentiation of adjacent hair-shaft layers. Notch4 is therefore a plausible candidate gene for AA. We have examined two polymorphisms in the coding sequence of the Notch4 gene at positions +1297 and +3063 in a case-control study of 116 AA patients and 142 ethnically matched, healthy control subjects. The initial analysis showed a significant association of AA in the overall data set with the Notch4(T+1297C) polymorphism (P<0.001) but not with Notch4(A+3063G). To confirm this association, we genotyped an additional 62 patients and found that the risk for disease was higher in Notch4(+1297C) homozygotes [odds ratio (OR) 3.43 (1.63, 7.19)] than in heterozygotes [OR 2.58 (1.57, 4.24)]. On classifying the patients by severity of disease, the association appeared to be confined to the severest form (alopecia universalis) [OR 4.02 (1.64, 9.88), P=0.0014]. These results support previous findings showing that different HLA susceptibility alleles are associated with mild and severe AA. PMID- 12589428 TI - Biallelic expression of HRAS and MUCDHL in human and mouse. AB - At least eight genes clustered in 1 Mb of DNA on human chromosome (Chr) 11p15.5 are subject to parental imprinting, with monoallelic expression in one or more tissues. Orthologues of these genes show conserved linkage and imprinting on distal Chr 7 of mice. The extended imprinted region has a bipartite structure, with at least two differentially methylated DNA elements (DMRs) controlling the imprinting of two sub-domains. We previously described three biallelically expressed genes ( MRPL23, 2G7 and TNNT3) in 100 kb of DNA immediately downstream of the imprinted H19 gene, suggesting that H19 marks one border of the imprinted region. Here we extend this analysis to two additional downstream genes, HRAS and MUCDHL (mu-protocadherin). We find that these genes are biallelically expressed in multiple fetal and adult tissues, both in humans and in mice. The mouse orthologue of a third gene, DUSP8, located between H19 and MUCDHL, is also expressed biallelically. The DMR immediately upstream of H19 frequently shows a net gain of methylation in Wilms tumors, either via Chr 11p15.5 loss of heterozygosity (LOH) or loss of imprinting (LOI), but changes in methylation in CpG-rich sequences upstream and within the MUCDHL gene are rare in these tumors and do not correlate with LOH or LOI. These findings are further evidence for a border of the imprinted region immediately downstream of H19, and the data allow the construction of an imprinting map that includes more than 20 genes, distributed over 3 Mb of DNA on Chr 11p15.5. PMID- 12589429 TI - The interleukin-6 (-174) G/C promoter polymorphism is associated with type-2 diabetes mellitus in Native Americans and Caucasians. AB - Chronic low-grade activation of the immune system may play a role in the pathogenesis of type-2 diabetes mellitus (T2DM). Interleukin-6 (IL6), a powerful inducer of hepatic acute phase response, has been implicated in the etiology of insulin resistance and T2DM. Recently, an IL6 promoter polymorphism (G/C) at position -174 was found to be associated with measures of insulin sensitivity. Because we have previously found an association between high IL6 levels and insulin resistance in both Pima Indians - a population with high rates of insulin resistance and T2DM - and Caucasians, we aimed to assess whether the IL6 promoter polymorphism is associated with T2DM in these populations. We genotyped the IL6 ( 174) G/C polymorphism using pyrosequencing in 463 Native Americans and by PCR RFLP in 329 Spanish Caucasians. Among the Spanish Caucasian subjects, there was a significant difference in genotypic distribution between diabetic and non diabetic subjects (P=0.028); the GG genotype was more common in diabetic (0.40) than in non-diabetic (0.29) subjects. The G allele was much more frequent in the Native American sample, and among a sample of 143 cases and 145 controls, the GG genotype was significantly more common in diabetic subjects (P=0.019). When this sample population was stratified according to ethnic heritage, all 211 subjects who were of full Pima Indian heritage had the GG genotype, whereas in the 77 American Indian subjects with non-Pima admixture, T2DM was associated with IL6 genotype (P=0.001). These findings are consistent with a role for genetic determinants of inflammation in the development of T2DM in both Native Americans and Caucasians. PMID- 12589431 TI - The ATPase ClpX is conditionally involved in the morphological differentiation of Streptomyces lividans. AB - ATP-dependent proteases of the ClpP type are widespread in eubacteria. These proteolytic complexes are composed of a proteolytic subunit and an ATPase subunit. They are involved in the degradation of denatured proteins, but also play a role in specific regulatory pathways. In Streptomyces lividans strains which lack the proteolytic subunit ClpP1, cell cycle progression has been shown to be blocked at early stages of growth. In this study, we examined the role of the ATPase subunit ClpX, a possible partner of the products of the clpP1 operon. A clpX mutant was obtained and it was shown that its growth was impaired only on acidic medium. Thus, the clpX phenotype differs from the clpP1 phenotype, indicating that these two components have only partially overlapping roles. We also analyzed the expression of clpX. Although clpX expression is increased under heat-shock conditions in many bacteria, we found that this is not the case in S. lividans. PMID- 12589432 TI - The 79,370-bp conjugative plasmid pB4 consists of an IncP-1beta backbone loaded with a chromate resistance transposon, the strA-strB streptomycin resistance gene pair, the oxacillinase gene bla(NPS-1), and a tripartite antibiotic efflux system of the resistance-nodulation-division family. AB - Plasmid pB4 is a conjugative antibiotic resistance plasmid, originally isolated from a microbial community growing in activated sludge, by means of an exogenous isolation method with Pseudomonas sp. B13 as recipient. We have determined the complete nucleotide sequence of pB4. The plasmid is 79,370 bp long and contains at least 81 complete coding regions. A suite of coding regions predicted to be involved in plasmid replication, plasmid maintenance, and conjugative transfer revealed significant similarity to the IncP-1beta backbone of R751. Four resistance gene regions comprising mobile genetic elements are inserted in the IncP-1beta backbone of pB4. The modular 'gene load' of pB4 includes (1) the novel transposon Tn 5719 containing genes characteristic of chromate resistance determinants, (2) the transposon Tn 5393c carrying the widespread streptomycin resistance gene pair strA-strB, (3) the beta-lactam antibiotic resistance gene bla(NPS-1) flanked by highly conserved sequences characteristic of integrons, and (4) a tripartite antibiotic resistance determinant comprising an efflux protein of the resistance-nodulation-division (RND) family, a periplasmic membrane fusion protein (MFP), and an outer membrane factor (OMF). The components of the RND-MFP OMF efflux system showed the highest similarity to the products of the mexCD-oprJ determinant from the Pseudomonas aeruginosa chromosome. Functional analysis of the cloned resistance region from pB4 in Pseudomonas sp. B13 indicated that the RND-MFP-OMF efflux system conferred high-level resistance to erythromycin and roxithromycin resistance on the host strain. This is the first example of an RND MFP-OMF-type antibiotic resistance determinant to be found in a plasmid genome. The global genetic organization of pB4 implies that its gene load might be disseminated between bacteria in different habitats by the combined action of the conjugation apparatus and the mobility of its component elements. PMID- 12589433 TI - Mkp1 and Mkp2, two MAPKAP-kinase homologues in Schizosaccharomyces pombe, interact with the MAP kinase Sty1. AB - Mkp1 ( MAPKAP kinase Schizosaccharomyces pombe 1) and Mkp2 are two members from fission yeast of the sub-class of putative MAPK-activated protein kinases in yeasts, the other known members being Rck1 and Rck2 from Saccharomyces cerevisiae. The Mkp1 protein is readily co-immunoprecipitated with Sty1 from S. pombe extracts; Mkp2 shows a weaker interaction with Sty1. In mkp1 mutants, conjugation and meiosis proceed more readily and rapidly than in wild-type cells, in analogy to what was previously found for S. cerevisiae rck1 mutants. Conversely, overexpression of mkp1(+) delays meiosis. Mkp1 is phosphorylated in vivo in a sty1(+)-dependent manner; this modification is removed when cells are starved for nitrogen, a condition that is conducive to entry into stationary phase and meiosis. Overexpression of mkp1(+), like a sty1 mutation, also causes vegetative cells to elongate. The level of Mkp1 phosphorylation drops as cells enter mitosis. We have localised Mkp1 to the cytoplasm, excluded from the nucleus, in vegetative cells. The Mkp1 protein accumulates in zygotic asci and is concentrated within spores. The mkp2(+) gene has no noticeable impact on meiosis. Mkp2 is excluded from the nucleus in vegetative cells, and is concentrated at the septa of dividing cells. Mkp2 does not accumulate in meiotic cells. PMID- 12589434 TI - Analysis of the petunia MADS-box transcription factor family. AB - Transcription factors are key regulators of plant development. One of the major groups of transcription factors is the MADS-box family, of which at least 80 members are encoded in the Arabidopsis genome. In this study, 23 members of the petunia MADS-box transcription factor family were investigated by Northern hybridisation, phylogenetic and yeast two-hybrid analyses. Many of the genes characterised appeared to have one or more close relatives that shared similar expression patterns. Comparison of the binding interactions of these proteins revealed that some show similar interaction patterns, and hence are likely to be functionally redundant. From an evolutionary point of view, their coding genes are probably derived from a recent duplication event. Furthermore, protein protein interaction patterns, in combination with expression patterns and phylogenetic classification, appear to offer good criteria for the identification of functional homologues. Based on comparison of such data between petunia and Arabidopsis, functions can be predicted for several MADS-box transcription factors in both species. PMID- 12589435 TI - Identification and characterisation of regions in the cellular protein LaXp180 and the Listeria monocytogenes surface protein ActA necessary for the interaction of the two proteins. AB - The Listeria monocytogenes surface protein ActA is an important virulence factor that plays an essential role in intracellular movement of Listeria cells by inducing actin polymerisation. The ActA protein is known to interact with several mammalian proteins including the phosphoprotein VASP, actin and the Arp2/3 complex. In a search for additional ActA-binding proteins we recently employed the yeast two-hybrid system to search for proteins that interact with ActA, and identified, among others, the mammalian protein LaXp180 as a binding partner. In the present study the interaction of the two proteins was investigated in more detail. A number of variants were tested in the yeast two-hybrid system for their ability to interact. On the basis of these assays, the 14 C-terminal amino acids of LaXp180 were identified as being necessary for the interaction with ActA. The proline-rich repeat (PRR) region of ActA was found to be necessary for the interaction with LaXp180, but upstream or downstream sequences are also required to enhance the specificity of the interaction. The second and third repeats in ActA are especially important, and the minimal sequence of ActA capable of interacting with LaXp180 was a proline- and glutamate-rich stretch of PRR3 fused to part of the N-terminal sequence of ActA. Further analysis using site-specific mutations located in either the C-terminal region of LaXp180 or the proline-rich motif of PRR3 of ActA showed that three positively charged amino acids in LaXp180 and two negatively charged amino acids in ActA are critical for the interaction of the two proteins. PMID- 12589436 TI - Amino acid residues involved in reversible thiol formation and zinc ion binding in the Streptomyces reticuli redox regulator FurS. AB - Streptomyces reticuli produces a mycelium-associated enzyme, CpeB, whose N terminal and C-terminal portions mediate heme-dependent catalase-peroxidase and heme-independent manganese-peroxidase activities, respectively. The regulator FurS governs transcription of the furS- cpeB operon. The thiol form of FurS contains one zinc ion per monomer and binds in this state to its cognate operator. Oxidation of SH groups within FurS induces the release of the zinc ion. Substitution of the codons for the amino acids cysteine 96, histidine 92 and 93, and tyrosine 59 in furS disrupts the in vivo repressor activity of FurS and results in enhanced synthesis of CpeB in corresponding S. lividans transformants. Biochemical and footprinting studies with FurS and its mutant derivatives revealed that the cysteine residues 96 and 99 are involved in reversible S-S bond formation, while cysteine 96 and the histidine residues 92 and 93 are required for zinc coordination, and tyrosine 59 is necessary for the binding of FurS to DNA. On the basis of these data, functional predictions can be made for the mycobacterial regulator FurA, a close homologue of FurS. PMID- 12589437 TI - Diversity of group II introns in the genome of Sinorhizobium meliloti strain 1021: splicing and mobility of RmInt1. AB - The number and diversity of known group II introns in eubacteria are continually increasing with the addition of new data from sequencing projects, but the significance of these introns in the evolution of bacterial genomes is unknown. We analyzed the main features of the group II introns present in the genome of the soil microorganism Sinorhizobium meliloti (strain 1021), the nitrogen-fixing symbiont of alfalfa, the DNA sequence of which was recently determined. Strain 1021 harbors three different classes of group II introns: RmInt1, of bacterial class D; SMb2147/SMb21167, which cluster within bacterial class C; and SMa1875, the phylogenetic class of which is uncertain. The group II introns SMb2147/SMb21167 and SMa1875 are widely distributed in S. meliloti, but are present in lower copy numbers than RmInt1. Strain 1021 harbors three copies of RmInt1, which is pSym-specific. Although RmInt1 is spliced in strain 1021, mobility assays suggested that, in contrast to other S. meliloti strains, the genetic background of strain 1021 does not support intron homing events. PMID- 12589438 TI - Structure, organization and expression of the genes encoding mitochondrial cytochrome c(1) and the Rieske iron-sulfur protein in Chlamydomonas reinhardtii. AB - The sequence and organization of the Chlamydomonas reinhardtii genes encoding cytochrome c(1) ( Cyc1) and the Rieske-type iron-sulfur protein ( Isp), two key nucleus-encoded subunits of the mitochondrial cytochrome bc(1) complex, are presented. Southern hybridization analysis indicates that both Cyc1 and Isp are present as single-copy genes in C. reinhardtii. The Cyc1 gene spans 6404 bp and contains six introns, ranging from 178 to 1134 bp in size. The Isp gene spans 1238 bp and contains four smaller introns, ranging in length from 83 to 167 bp. In both genes, the intron/exon junctions follow the GT/AG rule. Internal conserved sequences were identified in only some of the introns in the Cyc1 gene. The levels of expression of Isp and Cyc1 genes are comparable in wild-type C. reinhardtii cells and in a mutant strain carrying a deletion in the mitochondrial gene for cytochrome b (dum-1). Nevertheless, no accumulation of the nucleus encoded cytochrome c(1) or of core proteins I and II was observed in the membranes of the respiratory mutant. These data show that, in the green alga C. reinhardtii, the subunits of the cytochrome bc(1) complex fail to assemble properly in the absence of cytochrome b. PMID- 12589439 TI - Efficient disruption of a polyketide synthase gene ( pks1) required for melanin synthesis through Agrobacterium-mediated transformation of Glarea lozoyensis. AB - Glarea lozoyensis produces pneumocandin B(0), a potent inhibitor of fungal glucan synthesis. This industrially important filamentous fungus is slow-growing, is very darkly pigmented, and has not been easy to manipulate genetically. Using a PCR strategy to survey the G. lozoyensis genome for polyketide synthase (PKS) genes, we have identified pks1, a gene that consists of five exons interrupted by four introns of 56, 400, 50 and 341 bp. It encodes a 2124-amino acid protein with five catalytic modules: ketosynthase, acyltransferase, two acyl carrier sites, and thioesterase/Claisen cyclase. The transcriptional initiation and termination sites were found 43 bp upstream of the translational start codon and 295 bp downstream of the translational stop codon, respectively. Cluster analysis of 37 fungal ketosynthase modules grouped the Pks1p with PKSs involved in the biosynthesis of 1,8-dihydroxynaphthalene melanin. Disruption of pks1 yielded knockout mutants that displayed an albino phenotype, suggesting that pks1 encodes a tetrahydroxynaphthalene synthase. Gene replacement was achieved by Agrobacterium-mediated transformation, which proved to be simple and efficient. Loss of pigmentation occurred in more than half the transformants, and examination of six non-pigmented transformants showed that the functional genomic copy of the pks1 gene had been replaced by the disruption cassette in each case. A putative 1215-bp ORF (dsg) devoid of introns was present downstream from pks1. BLAST analysis of the 405-amino acid sequence of its predicted product showed a high degree of similarity with Zn(II)(2)Cys(6) binuclear cluster DNA-binding proteins, a class of fungal transcription factors involved in the regulation of polyketide production and other pathways. PMID- 12589440 TI - Chromosomal mapping of Brassica oleracea based on ESTs from Arabidopsis thaliana: complexity of the comparative map. AB - Expressed sequence tags (ESTs) from the Arabidopsis thaliana sequencing project were used to construct a genetic RFLP map for Brassica oleracea. Of the 110 A. thaliana ESTs tested, 95 were found to be informative RFLP probes in map construction. In total, 212 new loci corresponding to the 95 ESTs were added to the existing genetic map of B. oleracea. The enriched map covers all nine basic linkage groups and confirms that the chromosomes of B. oleracea and A. thaliana are similar in linear organization. However, varying levels of sequence conservation between the chromosomes of B. oleracea and A. thaliana were detected in different regions of the genomes. Long conserved regions encompassing entire chromosome arms in both genomes were identified; these are probably shared by descent. On the other hand, extensive rearrangements were observed in numerous chromosome regions, producing a mosaic of A. thaliana-like segments in the genome of Brassica. The presence of extensive chromosome duplication in A. thaliana was taken into consideration in the construction of the comparative maps of B. oleracea and A. thaliana. PMID- 12589441 TI - Construction of a BAC library of Rosa rugosaThunb. and assembly of a contig spanning Rdr1, a gene that confers resistance to blackspot. AB - A BAC library to serve as a general tool for the physical mapping and positional cloning of rose genes has been constructed from Rosa rugosa DNA. With 27,264 clones the library contains 5.2 genome equivalents. The library was used to assemble a contig of BAC clones spanning Rdr1, a locus that confers resistance to blackspot. For this purpose fine-scale mapping of the target locus was achieved by bulked segregant analysis using 816 AFLP primer combinations. The target region around Rdr1 comprises about 400 kb and is covered by a minimum of six BAC clones. Furthermore, the detection of at least five resistance gene analogs of the TIR-NBS-LRR family on the contig indicates the presence of a cluster of resistance genes around Rdr1. These results will not only allow the isolation and identification of Rdr1 in the near future, but also provide the tools for the physical mapping and positional cloning of other horticulturally interesting genes in roses. PMID- 12589442 TI - Deletion of Aspergillus nidulans aroC using a novel blaster module that combines ET cloning and marker rescue. AB - Blaster cassettes are of significant value in functional genomics, as they represent tools with which to inactivate duplicated or homologous genes in an individual organism. We have constructed a novel blaster module which allows repeated gene deletion in the filamentous fungus Aspergillus nidulans. Because bacterial resistance marker cassettes are employed as flanking repeats in direct orientation, the blaster cassette is suited for recombinogenic engineering by ET cloning in Escherichia coli. The functionality of the blaster module was demonstrated by deleting the chorismate mutase-encoding gene aroC of A. nidulans, followed by marker rescue based on mitotic recombination. The resulting aroCDelta strains are auxotrophic for phenylalanine but not tyrosine, and display a limited capacity for fruit body formation and ascosporogenesis, which depends on the phenylalanine/tyrosine supply. The data support the notion that amino acid status has a strong impact on cleistothecium development in A. nidulans. PMID- 12589443 TI - Isolation and characterisation of a calnexin homologue, clxA, from Aspergillus niger. AB - We describe the isolation of a gene (clxA) encoding calnexin from laboratory and industrial strains of Aspergillus niger. Calnexin is a chaperone, which specifically recognises monoglucosylated glycoproteins in the endoplasmic reticulum, and is thus an essential component of the process that assesses the folded state of nascent secreted glycoproteins. Manipulation of chaperones has previously been adopted in attempts to overcome some of the problems associated with the secretion of heterologous proteins from filamentous fungi. The A. niger clxA gene encodes a 562-residue protein with strong homology to the calnexin of Schizosaccharomyces pombe. The clxAgene product complements a S. pombe cnx1 mutant. Motifs associated with genes controlled via the Unfolded Protein Response (UPR) were identified by sequence homology in the promoter of clxA. Steady-state levels of clxA mRNA were elevated in a strain expressing bovine prochymosin fused to the catalytic domain of glucoamylase. The ORF is punctuated by four introns, and contains two sets of four repeated peptide motifs that are characteristic of the calnexin family, together with a putative membrane-spanning domain. Deletion studies indicate that clxA is not an essential gene in A. niger. PMID- 12589444 TI - The clot gene of Drosophila melanogaster encodes a conserved member of the thioredoxin-like protein superfamily. AB - The conversion of pyruvoyl-H(4)-pterin to pyrimidodiazepine (PDA), which is an essential step in the biosynthesis of the red components of Drosophila eye pigments known as drosopterins, requires the products of the genes sepia and clot. While the product of sepia has been shown to correspond to the enzyme PDA synthase, the role of clot remains unknown, although the clot(1) allele was one of the first eye-color mutants to be isolated in Drosophila melanogaster,and much genetic and biochemical data has become available since. Here we report the cloning of the clot gene, describe its molecular organization and characterize the sequence alterations associated with the alleles cl(1) and cl(2). The coding properties of the gene show that it encodes a protein related to the Glutaredoxin class of the Thioredoxin-like enzyme superfamily, conserved members of which are found in human, mouse and plants. We suggest that the Clot protein is an essential component of a glutathione redox system required for the final step in the biosynthetic pathway for drosopterins. PMID- 12589445 TI - Knowledge of word length does not constrain word identification. AB - Use of word length for word identification was examined in three naming experiments and one sentence reading experiment in which a foveally presented cue either matched or mismatched the length of a subsequently presented target word. Properties of the target were also manipulated so that it was either a high- or low-frequency word or so that its exterior letters were either consistent with a large or small pool of candidate words. The experiments converged in showing that effects of prior word length cuing were either negligible or absent. The word frequency of the target influenced its recognition but not the lexical constraint of its exterior letters. Importantly, effects of target word properties did not interact with effects of prior target length cuing. Together, these results indicate that knowledge of a word's spatial properties does not constrain its lexical processing. PMID- 12589446 TI - Planning of saccadic eye movements. AB - Most theories of the programming of saccadic eye movements (SEM) agree that direction and amplitude are the two basic dimensions that are under control when an intended movement is planned. But they disagree over whether these two basic parameters are specified separately or in conjunction. We measured saccadic reaction time (SRT) in a situation where information about amplitude and direction of the required movement became available at different moments in time. The delivery of information about either direction or amplitude prior to another reduced duration of SRT demonstrated that direction and amplitude were specified separately rather than in conjunction or in a fixed serial order. All changes in SRT were quantitatively explained by a simple growth-process (accumulator) model according to which a movement starts when two separate neural activities, embodying the direction and amplitude programming, have both reached a constant threshold level of activity. Although, in isolation, the amplitude programming was faster than the direction programming, the situation reversed when two dimensions had to be specified at the same time. We conclude that beside the motor maps representing the desired final position of the eye or a fixed movement vector, another processing stage is required in which the basic parameters of SEM, direction and amplitude, are clearly separable. PMID- 12589447 TI - Increasing the distance of an external focus of attention enhances learning. AB - Previous studies (e.g., Wulf, Hoss, & Prinz, 1998) have shown that motor learning can be enhanced by directing performers' attention to the effects of their movements ("external focus"), rather than to the body movements producing the effect ("internal focus"). The purpose of the present study was to test the hypothesis that increasing the distance between the body and the action effects might further enhance the learning advantages associated with an external focus of attention. The distance of the external effect was manipulated by instructing three groups of participants learning to balance on a stabilometer to focus on markers attached to the platform located at different distances from their feet. Specifically, two groups were to focus on distant markers on the outside ("far outside") or inside ("far-inside") of the platform, respectively, whereas another group was instructed to focus on markers close to their feet ("near"). In a retention test administered after two days of practice, all three external-focus groups showed generally more effective balance learning than an internal-focus control group. In addition, the far-outside and far-inside groups demonstrated similar performances, and both were more effective than the near group. Furthermore, the far-outside and far-inside groups showed higher-frequency movement adjustments than the near group. These results suggest that focusing on more distant effects results in enhanced learning by promoting the utilization of more natural control mechanisms. The findings are in line with a "constrained action" hypothesis that accounts for the relatively poorer learning associated with an attentional focus directed towards effects in close proximity to the body, or towards the body itself. PMID- 12589448 TI - Stroop dilution but not word-processing dilution: evidence for attention capture. AB - Stroop dilution refers to the observation that the impact of a color word on the naming of a color bar is reduced if another word-like object is displayed simultaneously. Recently, Brown, Roos-Gilbert, and Carr (1995) concluded that Stroop dilution is due to early-visual interference. This conclusion was evaluated in three experiments. Experiment 1 showed that, contrary to the predictions of an early-visual interference account, (a) diluters that are similar in terms of visual complexity induced different amounts of dilution and (b) the size of the dilution effect is proportional to the size of the Stroop interference effect when the diluters are used as single distractors. Experiments 2 and 3 revealed that when the position of the color bar is pre-cued, Stroop dilution disappears. We argue that these findings support Van der Heijden's (1992) attention-capture account of Stroop dilution. PMID- 12589449 TI - The role of instructions, practice, and stimulus-hand correspondence on the Simon effect. AB - Numerous studies of two-choice reaction tasks, including auditory and visual Simon tasks (i.e., tasks in which stimulus location is irrelevant) and visual compatibility tasks, have found that only spatial stimulus-response (S-R) correspondence affected S-R compatibility. Their results provided no indication that stimulus-hand correspondence was a significant factor. However, Wascher et al. (2001) suggested that hand coding plays a role in visual and auditory Simon tasks when the instructions are in terms of the finger/hand used for responding. The present experiments examined whether instructing subjects in terms of response locations or fingers/hands influenced the Simon effect for visual and auditory tasks. In Experiments 1-3, only spatial S-R correspondence contributed significantly to the Simon effect, even when the instructions were in terms of the fingers/hands. However, in Experiment 4, which used auditory stimuli and finger/hand instructions, the contribution of stimulus-hand correspondence increased with practice. PMID- 12589450 TI - Bimanual cross-talk during reaching movements is primarily related to response selection, not the specification of motor parameters. AB - Simultaneous reaching movements made with the two hands can show a considerable increase in reaction time (RT) when they differ in terms of direction or extent, compared to when the movements involve the same direction and extent. This cost has been attributed to cross-talk in the specification of the motor parameters for the two hands. However, a recent study [Diedrichsen, Hazeltine, Kennerley, & Ivry, (2001). Psychological Science, 12, 493-498] indicates that when reaching movements are cued by the onset of the target endpoint, no compatibility effects are observed. To determine why directly cued movements are immune from interference, we varied the stimulus onset asynchrony for the two movements and used different combinations of directly cued and symbolically cued movements. In two experiments, compatibility effects were only observed when both movements were symbolically cued. No difference was found between compatible and incompatible movements when both movements were directly cued or when one was directly cued and the other was symbolically cued. These results indicate that interference is not related to the specification of movement parameters but instead emerges from processes associated with response selection. Moreover, the data suggest that cross-talk, when present, primarily shortens the RT of the second movement on compatible trials rather than lengthening this RT on incompatible trials. PMID- 12589451 TI - [The molecular mechanisms of neovascular age-related macular degeneration]. AB - Age-related macular degeneration is the leading cause of irreversible vision loss in industrialized countries. While early forms of this disease with drusen and focal pigment alterations generally do not lead to relevant functional limitations, later forms of the disease, either through atrophy or choroidal neovascularization, are associated with significant visual impairment. A significant increase in knowledge about the molecular mechanisms of new vessel formation from the choriocapillaries has occurred over the past few years. This has already allowed for the clinical testing of pharmacological agents which inhibit the formation of new vessels in AMD. This article describes current research in the pathophysiology of choroidal neovascularization secondary to age related macular degeneration. PMID- 12589452 TI - [Pharmacological therapy for age-related macular degeneration. Current developments and perspectives]. AB - Current therapeutic options for age-related macular degeneration are very limited and are, at best, only capable of slowing visual loss. Because of this an intensive search for prophylactic agents capable of inhibiting the progression of this disease from early into late forms, as well as for new therapeutic approaches has been undertaken. While neuroprotective substances are hoped to prevent cellular death in this disease process, multiple substances capable of inhibiting neovascularization, such as VEGF inhibitors, are in clinical trials. Inhibitors of matrix-metalloproteinases (MMP) and chemotherapeutic agents are also being clinically tested as novel therapies for AMD. Other targets include the inhibition of toxic compound formation in lipofuscin granules such as AZ-E. What follows is an overview of different substances and their stages of development in clinical trials. PMID- 12589453 TI - [Angiogenesis research--quo vadis?]. AB - The field of angiogenesis research has seen an explosion of knowledge within the last 10 years. More than 3500 angiogenesis-related papers are presently being published per year compared to the less than 200 annual papers published in the early 1990s. Paralleling the progress in the field of basic angiogenesis research, translational research has led to the identification of more than 100 angiomanipulatory compounds. Presently, more than 40 substances are in various phases of clinical trials. The prospect of these exciting developments is presently dampened by the negative outcome of some advanced clinical trials. Thus, following euphoria and disillusion, the field is presently experiencing that translational clinical research requires endurance to eventually accomplish the successful implementation of angiomanipulatory therapies in the clinical setting. The present article provides an overview of the field of angiogenesis research and summarizes ongoing efforts aimed at developing angiomanipulatory therapies. PMID- 12589454 TI - [Transpupillary thermotherapy in exudative, age-related macular degeneration]. AB - OBJECTIVE: To assess the effectiveness of a single transpupillary thermotherapy (TTT) in patients with exudative, age-related macular degeneration (AMD). METHODS: In a prospective pilot study, 14 patients with a mean age of 78 years (range: 70-92 years) with subfoveal choroidal neovascularisation (CNV) due to exudative, age-related macular degeneration were treated with a single TTT using a diode laser (810 nm). Seven patients had a classic and seven an occult CNV. Laser beam size was 4.5 mm,the power setting was 800 mW and the exposure lasted 60 s. RESULTS: Twelve out of 14 patients could be followed for a period of 18 months. Stabilisation of the visual acuity was achieved in two patients, ten patients lost three or more Snellen lines. In none of the patients a regression of the CNV could be observed immediately after TTT. Inactive fibrotic scars developed in six patients at the end of the study. CONCLUSION: Our results suggest that TTT, according to our protocol, has no beneficial effect on the spontaneous course of the CNV in patients with AMD. PMID- 12589455 TI - [Retinal photoablation with the Erbium:YAG laser. Initial experimental results for traction-free removal of tissue]. AB - BACKGROUND: To investigate the potential of an Er:YAG laser for precise and traction-free removal of retinal layers in vitro. MATERIAL AND METHODS: Retinal ablation in porcine retinal explants was performed using a free running Er:YAG laser focused either into a low-OH quartz fiber or a sapphire fiber. The explants were treated under air or perfluorodecaline (PFD). The ablation depth was evaluated by optical coherence tomography (OCT) and histology sections. RESULTS: A radiant exposure of 5.0 J/cm(2) under air and PFD resulted in complete transsection of the neurosensory retina. Between 3.5 and 2.0 J/cm(2) the ablation depth and the defect patterns varied markedly and adjacent thermal zones areas were seen. Below 2.0 J/cm(2) no defects could be created in air, whereas under PFD the ablation extended into the ganglion cell layer. Ablations using a sapphire fiber and 2.0 J/cm(2) showed a significantly higher reproducibility of ablation depth, and homogeneous defect patterns limited to the nerve fiber layer could be produced without thermal damage. CONCLUSIONS: The Er:YAG laser system with a low-OH quartz fiber allowed ablation of inner retinal layers in vitro, but revealed a variable ablation depth and low reproducibility.However, a sapphire fiber showed markedly improved results. Therefore its use during vitreoretinal surgery seems possible not only as a cutting device but also as a tool for the ablation of fine retinal structures. PMID- 12589456 TI - [Stereotactic radiosurgery using the Gamma Knife for large uveal melanomas]. AB - BACKGROUND: We report the results over 3 years with stereotactic radiosurgery using the Gamma Knife for large and unsuitably located uveal melanomas. PATIENTS AND METHODS: A total of 100 patients (51 male, 49 female) have been treated since 1997 following a standardised treatment protocol (outpatient single-shot treatment, maximum dose 50 Gy, tumour margin dose min.25 Gy, retrobulbar anaesthesia alone for globe fixation). The localisation and/or dimension of the tumours did not allow radiation brachytherapy with Ru106 plaques. Of the tumours 18 were located in the ciliary body, 61 were located at the posterior pole, and 21 were located in the mid-periphery. All patients were followed and tested ophthalmologically and neuroradiologically at regular intervals. The 1-year follow-up data were available for 73 patients, 2-year follow-up data for 33 patients and 3-year follow-up-data for 17 patients. RESULTS: Before therapy the maximum apical tumour height (MAH) was median 7.8 mm (95% CI 2.9-12.5 mm): 1 year after treatment (73 patients) the MAH was median 5.7 mm (95% KI 2.4-10.2 mm),2 years after treatment (33 patients) the MAH was median 4.3 mm (95% KI 2.2-8.8 mm),and 3 years after treatment (17 patients) the MAH was median 4.6 mm (95% KI 2.4-8.5 mm). All differences to the MAH of the corresponding patients before treatment were statistically significant (paired t-test). Within the first year after treatment seven patients were enucleated due to a painful secondary glaucoma,within the second year after radiation two patients (one tumour recurrence, and one secondary glaucoma) and within the third year one more patient (tumour recurrence) was enucleated. CONCLUSIONS: Our 3-year results demonstrate that radiosurgery using the Gamma Knife is beneficial in achieving a local tumour control in 98% of eyes with large and unsuitably located uveal melanomas. The risk for a secondary enucleation is highest in the first year after treatment with a favourable overall rate of 10%. Due to the excellent local tumour control rate we decreased the maximum dose to 40 Gy (min.tumour margin dose 20 Gy) in the subsequently treated patients. PMID- 12589457 TI - [Piggyback lens implantation for anisometropia in pseudophakic eyes]. AB - OBJECTIVE: Long standing pseudophakia or previous laser capsulotomy make lens exchange for anisometropia or incorrect target refraction difficult. Piggyback lens implantation might be a less traumatic surgical alternative. PATIENTS AND METHODS: Eight patients had a secondary posterior chamber lens implant (PMMA) into the ciliary sulcus. Lens power was estimated by the preoperative refraction according to the refractive vergence formula and was controlled intraoperatively by an automatic refractometer. Surgical access minimized preoperative astigmatism. RESULTS: All patients had disturbing anisometropia of 4.57+/-2.86 D and had undergone cataract surgery 1-13 years previously. Piggyback lens implantation was combined with surgical aspiration of the secondary cataract in three patients. Due to recurrent PVR retinal detachment, one patient received a silicon oil refilling procedure and piggyback lens implantation. The preoperative spherical equivalent was -3.34+/-4.9 D. Mean follow up time was 16.6 months. Postoperative anisometropia was reduced to 1.1+/-1.01 D. The mean absolute deviation from target refraction was 0+/-0.92 D. Elevated IOP or secondary cataract formation in the lens interface were not observed. One patient developed pigment dispersion. CONCLUSION: Secondary piggyback lens implantation into the ciliary sulcus in front of an existing posterior chamber lens is a safe and effective refractive surgical procedure for special cases with predictable refraction. PMID- 12589458 TI - [Influence of long term occupational exposure to solvents on colour vision]. AB - BACKGROUND: The study was designed to determine the influence of chronic occupational exposition of organic solvent mixtures on colour vision of car painters. SUBJECTS AND METHODS: The 123 subjects (2 groups differing in organic solvents exposure and 1 control group) were examined using Ishihara-Panel, Lanthony Desaturated Panel D-15, Velhagen-Panel, and Tritan-Album. RESULTS: In the Velhagen-Panel 3% of the probands currently exposed to organic solvents, and 11% of formerly exposed probands developed a blue/yellow vision defect for the right eye. All control subjects perfectly finished this panel. In the Tritan Album 3% of currently exposed subjects and 26% of formerly exposed painters expressed a blue/yellow vision defect for the right eye, but also 7% of controls showed anomalies. Similar results were found for both panels with the left eye. The CCI difference in the D-15 test was significant between all three groups. CONCLUSION: The impaired colour vision may also be an important indicator of neuro-ophthalmological effects after long-term occupational exposure to organic solvents. PMID- 12589459 TI - [Immunochemical demonstration of human herpes virus 8 in conjunctival Kaposi's sarcoma]. AB - BACKGROUND: Angiomatous lesions of the conjunctiva and chronic conjunctivitis are frequently difficult to differentiate. CASE: A 28 year old white male sought help because of a conjunctival swelling. A biopsy made elsewhere had revealed a capillary haemangioma of the conjunctiva. Recently striking alterations of the skin led to the suspicion of a Kaposi's sarcoma. Immunohistochemical and dermatological examinations confirmed the diagnosis of a Kaposi's sarcoma and viral testing of serum revealed an HIV-infection. Besides intensive antiviral therapy, our patient underwent local irradiation of the right eye, the nose and of the left shoulder with an external beam. CONCLUSIONS: Kaposi's sarcoma is the most frequent Aids-associated malignant tumor. In about 30% of all patients with Aids, a Kaposi's sarcoma is diagnosed. The angiomatous character may lead to the histopathological diagnosis of a haemangioma. Human herpes virus 8 is known to be the transforming element. PMID- 12589460 TI - [Intravitreal chemotherapy for intraocular lymphoma]. AB - Primary CNS and intraocular non-Hodgkin's lymphoma is currently mostly treated with systemic chemotherapy. After initially successful tumor regression, recurrence is common and usually treated with radiotherapy. However, after good primary therapeutic response, new tumor growth is frequently observed. Because of actinic complications, radiotherapy can be applied only once. For an intraocular recurrence of primary CNS and intraocular lymphoma, intravitreal chemotherapy (0.4 mg methotrexate and 0.4 mg dexamethasone weekly for 4 weeks and once a month thereafter) can be performed. A few weeks after initiation, regression of intraocular tumors and eventually improvement of vision can be achieved. Eyes remained free of recurrence up to 2 years after initiation of intravitreal injections. Intravitreal chemotherapy is an effective treatment for ocular recurrence of primary CNS and intraocular lymphoma after systemic chemotherapy and radiotherapy. PMID- 12589461 TI - [Differences between clinical trials according to the German law on pharmaceuticals (Arzneimittelgesetz) and trials according to the German law on medical products (Medizinproduktegesetz)]. AB - PURPOSE: Similar to the registration process for pharmaceutical agents, medical devices have to undergo standardized clinical evaluation before being marketed and used in routine therapy or diagnostics. However, conduction, submission, and reporting of such clinical evaluations to authorities has to follow the German law on medical products(Medizinproduktegesetz,MPG), which in some central aspects differs remarkably from the German law on pharmaceuticals (Arzneimittelgesetz,AMG). METHODS: Relevant deviations of MPG requirements from those of the AMG are reviewed with particular emphasis on submission, conduction, and reporting of trials to the authorities in charge. RESULTS: Whereas AMG-based trials focus on the proof of efficacy of pharmaceutical agents, the MPG demands instead proof of functionality of the medical devices; the MPG therefore concentrates more on technically satisfactory results in the context of function and patient safety. The aim of MPG trials is thus CE marking instead of AMG-based registration. However, this focus on functionality implies that medical devices need not necessarily be tested in a clinical trial--in some settings evidence based evaluation alone will be sufficient. The decision on the necessity of a clinical trial is based mainly on the risk profile and invasive character of the device at hand. CONCLUSIONS: The early consideration of differences between AMG and MPG concerning the role and conduction of clinical trials will remarkably increase the (CE) certification process's outcome quality and juridical validity. PMID- 12589462 TI - [Amblyopia. Part 2: Precautionary measures and therapy]. PMID- 12589463 TI - Pre-mRNA splicing in Schizosaccharomyces pombe: regulatory role of a kinase conserved from fission yeast to mammals. AB - Most primary messenger RNA transcripts (pre-mRNAs) in eukaryotes contain intervening sequences that must be precisely removed to generate a functional mRNA. The excision of the intervening sequences, the introns, from a pre-mRNA and the concomitant joining of the flanking sequences, the exons, is called pre-mRNA splicing. Pre-mRNA splicing takes place in large ribonucleoprotein machinery, the spliceosome. Although the function and components of this machinery appear to be highly conserved between organisms, many distinct differences between budding yeast, Saccharomyces cerevisiae, and fission yeast, Schizosaccharomyces pombe, have been found, emphasizing their evolutionary distance. Most interestingly, fission yeast appears to reflect the more conservative evolutionary development regarding pre-mRNA splicing. Many spliceosomal components, including the five small nuclear RNAs, which most likely form the catalytic core of the spliceosome, show a higher degree of similarity with the components of the splicing machinery found in mammals. In addition, several regulatory components of the spliceosome detected in mammals are absent in Sac. cerevisiae, but present in Sch. pombe. Here, we review recent progress made in our understanding of the control of pre mRNA splicing in Sch. pombe. The focus is on Prp4p kinase, first discovered in fission yeast and also present in mammals, but absent in Sac. cerevisiae. Results from both mammals and Sch. pombe suggest that Prp4p plays a key role in regulating pre-mRNA splicing and in connecting this process with the cell cycle. PMID- 12589464 TI - Heterologous expression and characterization of Schizosaccharomyces pombe vacuolar carboxypeptidase Y in Saccharomyces cerevisiae. AB - To investigate the intracellular transport mechanism of the vacuolar carboxypeptidase of Schizosaccharomyces pombe (SpCPY), SpCPY was expressed in Saccharomyces cerevisiae and its biosynthesis and sorting were examined. When Sac. cerevisiae prc1Delta, devoid of intrinsic (Sc) CPY activity, was transformed with a plasmid carrying the Sch. pombe cpy1(+) gene, CPY activity was restored. Pulse-chase experiments revealed that SpCPY is initially synthesized in a pro precursor form and then converted to a heterodimer, the mature form, in Sac. cerevisiae cells. SpCPY was not processed into intermediate or mature forms in pep4 mutant cells, indicating that SpCPY was proteolytically cleaved in a PEP4 dependent manner in Sac. cerevisiae. Several vps mutants, which are defective in vacuolar protein-sorting, exhibited a defect in the maturation of SpCPY. Moreover, the maturation of SpCPY was severely inhibited in a vps10 strain, although the pro- segment of SpCPY does not contain a QRPL-like sequence, which is the putative targeting signal of ScCPY. When SpCPY was expressed in a wild type strain, more than 90% of ScCPY was normally sorted to the vacuole, indicating that SpCPY does not compete with ScCPY for vacuolar sorting. In contrast, expression of SpCPY resulted in a missorting of a ScCPY-invertase fusion protein to the cell surface. These results suggested that there are two different binding sites for SpCPY and ScCPY on Vps10p and that the binding of SpCPY to Vps10p interferes with the binding of a ScCPY-invertase fusion protein. PMID- 12589465 TI - Cooperative action of the NIT2 and NIT4 transcription factors upon gene expression in Neurospora crassa. AB - In Neurospora crassa, the nit-3 gene, which encodes nitrate reductase, an enzyme required for the utilization of inorganic nitrate, is subject to a high degree of genetic and metabolic regulation as a member of the nitrogen control circuit. The nit-3 gene promoter contains binding sites for a globally acting protein NIT2 and a pathway-specific protein NIT4. Expression of the nit-3 gene absolutely requires both the NIT2 and NIT4 transcription factors and only occurs under conditions of nitrogen source derepression and nitrate induction. In the sulfur control circuit, the cys-14 gene encodes sulfate permease II, which facilitates the assimilation of sulfate. Expression of cys-14 is strongly regulated by only a single positive-acting factor, CYS3. It was of interest to determine whether NIT2 or NIT4 alone was capable of turning on the expression of cys-14, since this structural gene is normally controlled by only one regulatory protein. NIT2- and/or NIT4-binding elements were introduced into the promoter of a wild-type cys 14 gene and these constructs were transformed into a cys-13(-) cys-14(-) mutant strain and into a nit-2(-) mutant host. We examined whether any of these cys-14 genes in these transformants could now be controlled as a nitrogen-regulated gene. Sulfate permease assays revealed that both NIT2 and NIT4 were required for cys-14 expression upon nitrate induction, while neither alone activated any detectable cys-14 expression. We thus conclude that neither NIT2 nor NIT4 is capable alone of activating gene expression in this context, but together they can cooperate to elicit strong activation. PMID- 12589466 TI - Evolution of multispecific mating-type alleles for pheromone perception in the homobasidiomycete fungi. AB - The evolution of multiple, independent and multispecific mating-type loci is a feature unique to homobasidiomycete fungi. To propose a model of evolution, data assembled for the wood-rotting fungus Schizophyllum commune were analyzed. In one mating-type locus, pheromone receptors and several pheromones are encoded which have been investigated in some detail and can be used to understand the ligand receptor interactions and activation of signal transduction which are essential to sexual propagation. Previous models for the evolution of new alleles were complicated and involved three subsequent steps (without selectable phenotype) prior to the establishment of a new stable pheromone-receptor pair. This paper presents a model for the evolution of new specificities by recombination and selection that incorporates the multi-state receptor activation recently established for S. commune, explaining differential responses to different pheromones in one receptor molecule. The model takes into account the occurrence of multiple pheromone genes in each locus and unilateral nuclear donor/acceptor strains that may in nature act as steps in the evolution of new specificities. A second homobasidiomycete fungus, Coprinus cinereus, was similarly characterized at the molecular level. Data acquired in this system support the conclusion that the presented model can be generalized. PMID- 12589467 TI - The little difference: in vivo analysis of pheromone discrimination in Schizophyllum commune. AB - The B mating type of Schizophyllum commune is defined by a multi-specific pheromone/receptor system. The interaction of pheromone receptors and their ligands, encoded by the Balpha locus, triggers sexual development. The receptors belong to the family of G protein-coupled seven-transmembrane-domain receptors, while the ligands are small lipopeptide pheromones. A productive interaction is only possible between molecules derived from different specificities. There is no induction of sexual development by pheromones of self-specificity. Since there are nine versions of different specificity for pheromones and receptors in Balpha, this system can be used to study multi-ligand discrimination. We investigated pheromone discrimination using chimeric receptor molecules and the influence of single point mutations on activation profiles of the receptor. PMID- 12589468 TI - Expression of a hypovirulence-causing double-stranded RNA is associated with up regulation of quinic acid pathway and down-regulation of shikimic acid pathway in Rhizoctonia solani. AB - We reported previously that a 3.6-kb double-stranded RNA, designated as M2, is associated with hypovirulence in the Rhizoctonia solani isolate Rhs 1A1 and proposed that the M2-encoded putative polypeptide A (pA) might interfere with the regulation of the quinate and shikimate pathways. In this study, Western blot analysis showed that a protein band of the predicted size (83 kDa) binds antibodies specific to a pA epitope and is detectable in M2-containing but not in M2-lacking cultures. A mRNA, associated with Rhs 1A1 polysomes immunoprecipitated with anti-pA antibodies, has a sequence basically identical to that of the sense strand of M2. The normally inducible quinate pathway was constitutively expressed, whereas the shikimate pathway was down-regulated in the M2-containing, hypovirulent Rhs 1A1. Finally, the relative concentration of phenylalanine, precursor of the virulence determinant phenylacetic acid, was correlated with the degree of pathogenicity in the virulent Rhs 1AP but not in the hypovirulent Rhs 1A1. PMID- 12589469 TI - Proteobacteria-like ferrochelatase in the malaria parasite. AB - A gene encoding the heme biosynthetic enzyme ferrochelatase (FC) was found in the genomic DNA databases of Plasmodium spp. The predicted amino acid sequence of malarial FC is highly conserved and fairly well conserved by comparison with other orthologues. The FC genes of P. falciparum and P. yoelii are transcribed and the mRNAs are processed to encode polypeptides of the expected amino acid sequence. The cloned cDNA for the FC of P. falciparum successfully rescued a FC null mutant of Escherichia coli, indicating that it encodes an active enzyme. Unlike eukaryotic FCs, the malarial enzyme lacks a characteristic extension at the C-terminus. In addition, the sequence of the malarial FC resembles proteobacterial orthologues rather than eukaryotic enzymes. Strikingly, the malarial FC lacks a bipartite presequence at its N-terminus, unlike delta aminolevulinic acid dehydratase of the same organism. This suggests an unusual intracellular distribution of heme biosynthetic enzymes, involving multiple subcellular compartments. PMID- 12589470 TI - Mitotic recombination in Saccharomyces cerevisiae. AB - Mitotic homologous recombination (HR) is an important mechanism for the repair of double-strand breakS and errors occurring during DNA replication. It is likely that the recombinational repair of DNA lesions occurs preferentially by sister chromatid exchanges that have no genetic consequences. However, most genetically detectable HR events occur between homologous DNA sequences located at allelic positions in homologous chromosomes, or between DNA repeats located at ectopic positions in either the same, homologous or heterologous chromosomes. Mitotic recombination may occur by multiple mechanisms, including double-strand break repair, synthesis-dependent strand annealing, break-induced replication and single-strand annealing. The occurrence of one recombination mechanism versus another depends on different elements, including the position of the homologous partner, the initiation event, the length of homology of the recombinant molecules and the genotype. The genetics and molecular biology of the yeast Saccharomyces cerevisiae have proved essential for the understanding of mitotic recombination mechanisms in eukaryotes. Here, we review recent genetic yeast data that contribute to our understanding of the different mechanisms of mitotic recombination and the in vivo role of the recombination proteins. PMID- 12589471 TI - Investigating the importance of proteasome-interaction for Rad23 function. AB - Rad23 contributes to diverse cellular functions that include DNA repair, stress response and growth control. An amino-terminal ubiquitin-like (UbL) domain in Rad23 interacts with catalytically active proteasomes and internal sequences bind multi-ubiquitinated proteins. Rad23 regulates the assembly of substrate-linked multi-ubiquitin chains, promotes efficient degradation of model substrates, and plays an overlapping role with the proteasome subunit, Rpn10. These and other results led to the hypothesis that Rad23 translocates proteolytic substrates to the proteasome to promote degradation. It was previously shown that the UbL domain in Rad23 could be functionally replaced by ubiquitin. However, monomeric ubiquitin does not bind the proteasome efficiently, and we therefore investigated whether proteasome interaction was required for all Rad23 functions. We report here that the ubiquitin moiety in Ub-rad23 is ubiquitinated in vivo and could provide an alternate mechanism for binding the proteasome. These results suggest that the localization of Rad23 to the proteasome, either by its UbL domain, or following ubiquitination of an amino-terminal ubiquitin moiety (Ub-rad23), is necessary for full activity. PMID- 12589472 TI - Impact of the cross-pathway control on the regulation of lysine and penicillin biosynthesis in Aspergillus nidulans. AB - The non-proteinogenic amino acid, alpha-aminoadipate, defines the biosynthetic branch-point of lysine and penicillin biosynthesis in the filamentous fungus, Aspergillus nidulans. Regulation of both pathways was analysed in response to amino acid limitation. The lysF-encoded homoaconitase acts upstream of the alpha aminoadipate branch point, whereas the lysA gene product, saccharopine dehydrogenase, catalyses the ultimate step of the lysine-specific branch. The lysA gene from A. nidulans was identified and isolated. Amino acid starvation resulted in significantly increased transcription of lysA but not lysF. Starvation-dependent changes in transcription levels of lysA were dependent on the presence of the central transcriptional activator of the cross-pathway control (CPCA). The effect of amino acid starvation under penicillin-producing conditions was analysed in A. nidulans strains with reporter genes for the penicillin-biosynthesis genes, acvA and ipnA, and genetically altered activity of the cross-pathway control. Overproduction of CPCA decreased expression of ipnAand acvA reporter genes and even more drastically reduced penicillin production. This work suggests that, upon amino acid starvation, the cross-pathway control overrules secondary metabolite biosynthesis and favours the metabolic flux towards amino acids instead of penicillin in A. nidulans. PMID- 12589473 TI - The QDE-3 homologue RecQ-2 co-operates with QDE-3 in DNA repair in Neurospora crassa. AB - The post-transcriptional gene silencing mechanism in Neurospora crassa, called quelling, was shown to involve the products of three genes termed quelling defective. A homologue to the qde-3 gene encoding a putative RecQ-type DNA helicase was isolated and was named RecQ-2. Characterisation of the RecQ-2 gene has revealed that it is not involved in quelling, but may co-operate with the qde 3 gene product in a pathway that repairs damage to DNA caused by the chemical mutagens methyl methanesulfonate and N-methyl- N'-nitro- N-nitrosoguanidine. These results indicate that the qde-3 RecQ helicase may have a dual role in N. crassa, either acting alone as an essential component of the quelling mechanism or together with the RecQ-2 RecQ helicase, as part of a process to repair DNA lesions during replication. PMID- 12589474 TI - Stable transformation of the oomycete, Phytophthora infestans, using microprojectile bombardment. AB - Germinated asexual sporangia, zoospores, and mycelia of Phytophthora infestans were transformed to G418-resistance by microprojectile bombardment. After optimization, an average of 14 transformants/shot were obtained, using 10(6) germinated sporangia and gold particles coated with 1 microg of vector. Transformants displayed tandem or simple insertions of vector sequences within chromosomes. Most primary transformants were heterokaryons of transformed and wild-type nuclei, a state which generally persisted for generations, even with G418 selection. Transgenic homokaryons were easily obtained from primary transformants through G418 selection of zoospores. To facilitate the optimization of transformation, experiments were performed using a vector containing neomycin phosphotransferase (npt) and beta-glucuronidase (GUS) genes fused to oomycete transcriptional regulatory sequences. To indicate which orientations of transgenes would maximize their expression, head-to-head, head-to-tail, or tail to-tail orientations of npt and GUS were compared. Each yielded similar rates of transformation and levels of GUS activity, indicating little transcriptional interference. PMID- 12589475 TI - Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants. AB - Protoplasts of the pathogenic plant fungus, Crinipellis perniciosa, were transformed to hygromycin B resistance using the pAN7-1 plasmid, which contains the Escherichia coli hph gene under the control of Aspergillus nidulans regulatory sequences. The pAN7-1 plasmid was introduced by PEG/CaCl(2) treatment. Transformation frequencies of 1.6-2.5 transformants/microg of DNA were achieved. About 54% of the transformants were abortive and 40 analyzed transformants were mitotically stable and showed different hygromycin B resistance levels. The presence of the hph gene was checked by PCR in five transformants and the integration of multiple plasmid copies into different genome sites was observed by Southern analysis. This is the first report of a C. perniciosa transformation system and represents an important step for further research into genetic manipulation of this fungal plant pathogen. PMID- 12589476 TI - Positron emission tomography with 11C-acetate and 18F-FDG in prostate cancer patients. AB - Visualisation of primary prostate cancer, its relapse and its metastases is a clinically relevant problem despite the availability of state-of-the-art methods such as CT, MRI, transrectal ultrasound and fluorine-18 fluorodeoxyglucose positron emission tomography ((18)F-FDG PET). The aim of this study was to evaluate the efficacy of carbon-11 acetate and (18)F-FDG PET in the detection of prostate cancer and its metastases. Twenty-five patients were investigated during the follow-up of primary prostate cancer, suspected relapse or metastatic disease using (11)C-acetate PET; 15 of these patients were additionally investigated using (18)F-FDG PET. Fourteen patients were receiving anti-androgen treatment at the time of the investigation. Lesions were detected in 20/24 (83%) patients using (11)C-acetate PET and in 10/15 (75%) patients using (18)F-FDG PET. Based on the results of both PET scans, one patient was diagnosed with recurrent lung cancer. Median (18)F-FDG uptake exceeded that of (11)C-acetate in distant metastases (SUV =3.2 vs 2.3). However, in local recurrence and in regional lymph node metastases, (11)C-acetate uptake (median SUVs =2.9 and 3.8, respectively) was higher than that of (18)F-FDG (median SUVs =1.0 and 1.1, respectively). A positive correlation was observed between serum PSA level and both (11)C-acetate uptake and (18)F-FDG uptake. (11)C-acetate seems more useful than (18)F-FDG in the detection of local recurrences and regional lymph node metastases. (18)F-FDG, however, appears to be more accurate in visualising distant metastases. There may be a role for combined (11)C-acetate/(18)F-FDG PET in the follow-up of patients with prostate cancer and persisting or increasing PSA. PMID- 12589477 TI - Assessment of clinical utility of 18F-FDG PET in patients with head and neck cancer: a probability analysis. AB - The purpose of this study was to calculate disease probabilities based on data of patients with head and neck cancer in the register of our institution and to perform a systematic review of the available data on the accuracy of PET in the primary assessment and follow-up of patients with head and neck cancer. The pre test probability of head and neck cancer among patients in our institutional data registry was assessed. Then the published literature was selected and appraised according to a standard protocol of systematic reviews. Two reviewers independently selected and extracted data on study characteristics, quality and accuracy. Accuracy data were used to form 2 x 2 contingency tables and were pooled to produce summary receiver operating characteristic (ROC) curves and summary likelihood ratios for positive and negative testing. Finally post-test probabilities were calculated on the basis of the pre-test probabilities of this patient group. All patients had cytologically or histologically proven cancer. The prevalence of additional lymph node metastases on PET in staging examinations was 19.6% (11/56), and that of locoregional recurrence on restaging PET was 28.6% (12/42). In the primary assessment of patients, PET had positive and negative likelihood ratios of 3.9 (2.56-5.93) and 0.24 (0.14-0.41), respectively. Disease probabilities were therefore 49.4% for a positive test result and 5.7% for a negative test result. In the assessment of recurrence these values were 3.96 (2.8 5.6) and 0.16 (0.1-0.25), resulting in probabilities of 49.7% and 3.8%. PET evaluation for involvement of lymph nodes had positive and negative likelihood ratios of 17.26 (10.9-27.3) and 0.19 (0.13-0.27) for primary assessment and 11.0 (2.93-41.24) and 0.14 (0.01-1.88) for detection of recurrence. The probabilities were 81.2% and 4.5% for primary assessment and 73.3% and 3.4% for assessment of recurrence. It is concluded that in this clinical setting the main advantage of PET is the ability to reliably rule out the presence of disease in both staging and restaging. Further research is required to derive probabilities for individual patients from sequential testing as applied in the diagnostic work-up of patients with head and neck cancer. PMID- 12589478 TI - Whole-body distribution and dosimetry of O-(2-[18F]fluoroethyl)-L-tyrosine. AB - The whole-body distribution of O-(2-[(18)F]fluoroethyl)- l-tyrosine (FET) was studied in seven patients with brain tumours by positron emission tomography (PET). Based on the IMEDOSE and MIRDOSE procedures, radiation absorbed doses were estimated from whole-body PET scans acquired approximately 70 and 200 min after i.v. injection of 400 MBq FET. After injection of FET, the peak of radioactivity in the blood was observed after 1.5 min, and a plateau of nearly constant radioactivity was reached at 20 min. The whole-body distribution of FET showed the highest activities in the urinary tract. All other organs exhibited only moderate FET uptake (SUV /=0.99). Segmental wall motion analysis revealed good agreement between gated FDG PET data and cMRI (kappa =0.62+/-0.03). In conclusion, despite small systematic differences which contributed mainly to the lower temporal resolution of gated FDG PET, agreement between gated FDG PET and cMRI was good across a wide range of volumes and LVEF values as well as for wall motion analysis. Therefore, gated FDG PET provides clinically relevant information on function and volumes, using the commercially available software package QGS. PMID- 12589482 TI - Dynamic gadolinium-enhanced MRI evaluation of porcine femoral head ischemia and reperfusion. AB - OBJECTIVE: To examine the potential of gadolinium (Gd)-enhanced dynamic MRI in the detection of early femoral head ischemia. Furthermore, to apply a three compartment model to achieve a clinically applicable MR index for femoral head perfusion during the steady state and arterial hip joint tamponade. DESIGN AND MATERIALS: In a porcine model femoral head perfusion was measured by radioactive tracer microspheres and by using a dynamic Gd-enhanced MRI protocol. Femoral head perfusion measurements and MRI tests were performed unilaterally before, during and after the experimentally induced ischemia of one of the hip joints. Ischemia was induced by increasing intra-articular pressure to 250 mmHg. RESULTS: All pigs showed ischemia of the femoral head epiphysis under hip joint tamponade followed by reperfusion to the same level as before joint tamponade. In two cases perfusion after removal of tamponade continued to be low. In dynamic MRI measurements increases in signal intensity were seen after intravenous infusion of Gd-DTPA, followed by a slow decrease in signal intensity. The signal-intensity curve during femoral head ischemia had a minor increase. Also the coefficient determined was a helpful indicator of femoral head ischemia. CONCLUSIONS: Femoral head blood flow as measured by microspheres fell significantly under joint tamponade. Early detection of this disturbed regional blood flow was possible using a dynamic MRI procedure. A biomathematical model resulted from the evaluation of the intervals of signal intensity over time which allows detection of bone blood flow changes at a very early stage. Using this new method earlier detection of femoral head necrosis may be possible. PMID- 12589483 TI - Vertebral chondroblastoma. AB - OBJECTIVE: To determine the age distribution, gender, incidence, and imaging findings of vertebral chondroblastoma, and to compare our series with findings from case reports in the world literature. DESIGN AND PATIENTS: Case records and imaging findings of nine histologically documented vertebral chondroblastomas were retrospectively reviewed for patient age, gender, vertebral column location and level, morphology, matrix, edema, soft tissue mass, spinal canal invasion, and metastases. Our findings were compared with a total of nine patients identified from previous publications in the world literature. The histologic findings in our cases was re-reviewed for diagnosis and specifically for features of calcification and secondary aneurysmal bone cyst (ABC). Clinical follow-up was requested from referring institutions. RESULTS: Nine of 856 chondroblastomas arose in vertebrae (incidence 1.4%; thoracic 5, lumbar 1, cervical 2, sacral 1). There were six males and three females ranging in age from 5 to 41 years (mean 28 years). Satisfactory imaging from seven patients revealed the tumor to arise from the posterior elements in four and the body in three. All tumors were expansive, six of seven were aggressive, and the spinal canal was significantly narrowed by bone or soft tissue mass in six. In one patient canal invasion was minimal. Calcification was pronounced in two and subtle in four. The sole nonaggressive appearing tumor was heavily mineralized. Bony edema and secondary ABC were not seen on MR imaging. None of the cases had microscopic features of significant secondary ABC. Calcification, and specifically "chicken wire" calcification, was identified in two patients. Pulmonary metastases occurred in none. CONCLUSIONS: Vertebral chondroblastoma is a rare neoplasm that presents later in life than its appendicular counterpart. On imaging it is aggressive in appearance with bone destruction, soft tissue mass, and spinal canal invasion. The lesions contain variable amounts of mineral. Secondary aneurysmal cyst bone formation was not a feature in our study group. PMID- 12589484 TI - Enlarged spinoglenoid notch veins causing suprascapular nerve compression. AB - OBJECTIVE: To report the magnetic resonance (MR) imaging findings of enlarged veins in the spinoglenoid notch as a cause of suprascapular nerve compression. DESIGN AND PATIENTS: Six patients presented to MR imaging for evaluation of chronic shoulder pain. Clinical information and MR imaging studies were reviewed. The spinoglenoid notch vascular structures were compared with measurements made in 10 age-matched controls. RESULTS: Spinoglenoid notch vascular structures measured in 10 asymptomatic age-matched control patients ranged from 1 to 4 mm in diameter with an average of 2.2 mm. The six study patients had vascular structures that ranged from 6 to 10 mm in diameter with an average of 8.4 mm. Atrophy and fatty infiltration of the infraspinatus muscle was noted as an associated finding at MR imaging in all six patients. Surgery was performed in three of the six patients, at which time a venous varix was identified in the spinoglenoid notch in all three patients. CONCLUSION: We describe distended veins in the spinoglenoid notch. These may be readily apparent at MR imaging and should be distinguished from paralabral ganglion cysts compressing the suprascapular nerve in the absence of labral tears, especially if percutaneous aspiration of a ganglion cyst is entertained. PMID- 12589485 TI - Intermittent dislocation of the flexor hallucis longus tendon. AB - Dislocation of the flexor hallucis longus tendon is an exceptional occurrence. To our knowledge, this is the first case ever reported of an intermittent dislocation in a 17-year-old woman; she was a synchronised swimmer. She consulted for a right internal retro-malleolar syndrome. Voluntary "snap" was triggered by a mechanism which combined maximal ankle dorsiflexion and interphalangeal plantar flexion of the toes. Non-enhanced dynamic helical CT and axial MRI were performed, which revealed the dislocation of the right flexor hallucis longus tendon outside the posterior intertubercular talar groove. Static and dynamic imaging would appear to be required to make this uncommon diagnosis. PMID- 12589486 TI - Calcific tendinitis of the rotator cuff as a cause of drooping shoulder. AB - We describe a case of inferior glenohumeral subluxation or drooping shoulder secondary to acute calcific tendinitis of the rotator cuff. The various etiologies of drooping shoulder and the specific causes determining glenohumeral widening in our report are discussed. The importance in recognizing this uncommon complication of a common abnormal finding and correction by aspiration is stressed. PMID- 12589487 TI - Progression from calcifying tendinitis to rotator cuff tear. AB - This report documents the clinical, radiographic and histologic findings in a 46 year-old man with calcifying tendinitis in his left shoulder which progressed to rotator cuff tear. The patient had a 1-year history of repeated calcifying tendinitis before being referred to our hospital. On the initial visit, radiographs and magnetic resonance imaging (MRI) revealed calcium deposition localized in the supraspinatus tendon without apparent tear. Three months after the first visit, MRI revealed a partial-thickness rotator cuff tear at the site of calcium deposition. Surgical and histologic findings demonstrated that calcium deposition was the cause of cuff rupture. To our knowledge, based on a review of the English literature, this is the first case report in which the progression from calcifying tendinitis to rotator cuff tear has been serially observed. PMID- 12589488 TI - Acetabular paralabral cyst: an uncommon cause of sciatica. AB - The association between tears of the acetabular labrum and paralabral cysts has been well documented, and magnetic resonance imaging (MRI) has been shown to be the most accurate noninvasive method of depicting not only the normal anatomic structures of the hip, but also the common pathologic processes such as labral tears and paralabral cysts. We present the case of an acetabular paralabral cyst that resulted in clinically symptomatic compression of the sciatic nerve. PMID- 12589489 TI - Palmar dislocation of the trapezoid. AB - Palmar dislocation of the trapezoid is a rare injury, which results from substantial trauma to the wrist. It is associated with other bony or ligamentous injuries in the wrist. If unrecognized, loss of function of the hand and early osteoarthritis may result. Treatment of choice is open reduction and internal fixation, which generally achieves good functional results. We describe a new case of palmar dislocation of the trapezoid and review the world literature with emphasis on the radiographic findings. PMID- 12589490 TI - Tenosynovial osteochondromatosis of the tarsal tunnel. AB - A case of tenosynovial osteochondromatosis in the tarsal tunnel in a 23-year-old man is presented. The lesion was treated surgically, and multiple osteochondromas were excised, which had no continuity with any tarsal bone or joint cavity but did with the sheaths of the flexor digitorum longus and flexor hallucis longus. Histologic examination of the lesion showed a fibrous capsule, hyaline cartilage and extensive areas of cancellous bone. Necrosis and mitosis were absent in the hyaline cartilage and there were no synovial nodules indicative of synovial metaplasia. The macroscopic findings showed "end-stage" tenosynovial osteochondromatosis. There was no evidence of recurrence 5 years after operation, and the patient remains free of symptoms. PMID- 12589491 TI - Cervical involvement in SAPHO syndrome: imaging findings with a 10-year follow up. AB - Osteoarticular manifestations of SAPHO syndrome include vertebral lesions, typically in the thoracic segment. Chronic inflammatory changes are well depicted by MRI. We report the imaging findings with a 10-year follow-up in a case of SAPHO syndrome with marked cervical lesions. PMID- 12589492 TI - Giant cell tumor of the rib. AB - A 27-year-old woman with a giant cell tumor of the rib with a cystic-hemorrhagic appearance underwent surgery consisting of en bloc resection and reconstruction of the thoracic wall with Marlex mesh, reinforced with two titanium plates. When possible this type of tumor requires resection, instead of radiotherapy, since the majority of cases of malignant transformation are linked to prior radiation therapy. PMID- 12589493 TI - A new syndrome of "spondylo-epi-metaphyseal dysplasia: mixed type". AB - A new type of rare bone dysplasia is described, which shares some common features with spondylo-meta-epiphyseal dysplasia: short limb-abnormal calcification type and lethal metatropic dysplasia. Besides these features, the present case has some additional unusual features. Facial malformation was very obvious and of a different type. The nose and nares were completely flattened. Hypertrophied acetabular bones, round densities on the ilia, premature ossification of many epiphyses and carpal bones, curvilinear calcifications in some joints, fusion of the ischiopubic rami, calcification of many costal cartilages and thick sclerotic base of the skull were a few of the significant findings. On the basis of the clinical and radiological features, the condition has been named "spondylo-epi metaphyseal dysplasia: mixed type". PMID- 12589494 TI - Bilateral elastofibroma of the thighs with concomitant subscapular lesions. AB - We report a case of elastofibroma that occurred in both thighs and subscapular regions. The MR signal intensity of the lesions was typical for elastofibroma. Although bilateral involvement of the thighs is exceptional, the presence of concomitant characteristic bilateral subscapular lesions was helpful in making the diagnosis of elastofibroma. PMID- 12589495 TI - Specialist or generalist? PMID- 12589496 TI - Multiplex PCR design strategy used for the simultaneous amplification of 10 Y chromosome short tandem repeat (STR) loci. AB - The simultaneous amplification of multiple regions of a DNA template is routinely performed using the polymerase chain reaction (PCR) in a process termed multiplex PCR. A useful strategy involving the design, testing, and optimization of multiplex PCR primer mixtures will be presented. Other multiplex design protocols have focused on the testing and optimization of primers, or the use of chimeric primers. The design of primers, through the close examination of predicted DNA oligomer melting temperatures ( T(m)) and primer-dimer interactions, can reduce the amount of testing and optimization required to obtain a well-balanced set of amplicons. The testing and optimization of the multiplex PCR primer mixture constructed here revolves around varying the primer concentrations rather than testing multiple primer combinations. By solely adjusting primer concentrations, a well-balanced set of amplicons should result if the primers were designed properly. As a model system to illustrate this multiplex design protocol, a 10 loci multiplex (10plex) Y chromosome short tandem repeat (STR) assay is used. PMID- 12589497 TI - HPLC assay for methylmalonyl-CoA epimerase. AB - Methylmalonyl-CoA epimerase (MCE) is broadly distributed in nature and has diverse cellular roles. Many MCE homologues are represented in public databases, but the biochemical function and physiological roles of the majority of these putative proteins have not been investigated. Here, a simplified assay for MCE is described. In this assay, MCE converted (2S)-methylmalonyl-CoA to (2R) methylmalonyl-CoA which in turn was converted to succinyl-CoA by methylmalonyl CoA mutase, an enzyme specific for the 2 R isomer. MCE activity was quantified by measuring the disappearance of methylmalonyl-CoA by HPLC. To obtain the methylmalonyl-CoA mutase which was required as a reagent for the assay, an Escherichia coli strain was constructed that expressed high levels of this enzyme as a fusion protein with an 8x histidine tag. This allowed purification of the mutase in a single affinity chromatography step. Previously reported MCE assays required radioactive substrates and/or multiple reagent enzymes that were difficult to obtain. The assay reported here overcomes these difficulties and hence will facilitate studies of MCEs. Such enzymes play important roles in the metabolism of both prokaryotes and higher eukaryotes including humans. PMID- 12589498 TI - Gramicidin A interaction at a dioleoyl phosphatidylcholine monolayer on a mercury drop electrode. AB - A biosensor where the sensing surface is a fluid dioleyl phosphatidylcholine monolayer (DOPC) deposited on a mercury drop was used. The lipid monolayer was held in 0.1 M NaCl and a concentration of gramicidin A in the range 0-12 nM was used. Electrochemical impedance spectroscopy in the frequency range 0.1-65 kHz was employed to investigate how the defect-free monolayer responds to interactions of gramicidin A in solution. The data was analyzed both with multivariate data analysis and classical electrochemical methods. The principal component analysis of the resulting impedance spectra gave a linear dependence on the concentration of gramicidin A. An increasing permittivity was observed in the low-frequency regime with increasing concentration of gramicidin A in solution. PMID- 12589499 TI - Determination of siloxanes, silicon, and platinum in tissues of women with silicone gel-filled implants. AB - Silicone [poly(dimethylsiloxane)] gel used in breast implants has been known to migrate through intact silicone elastomer shells, resulting in the clinically observable "gel bleed" on the implant surface. Although silicon concentrations in capsular tissues of women with silicone prostheses have been measured with element-specific silicon analyses, no silicone-specific investigation of these tissues has been performed as yet.A combination of element-specific inductively coupled plasma high-resolution isotope dilution mass spectrometry (ICP-HR-IDMS) and species-specific gas chromatography coupled mass spectrometry (GC-MS) was used to analyze silicon, platinum, and siloxanes in prosthesis capsule, muscle, and fat tissues of women (n=3) who had silicone gel-filled breast implants and in breast tissue of non-augmented women (n=3) as controls.In all tissues of augmented women, siloxanes, in particular octamethylcyclotetrasiloxane (D4), decamethylcyclopentasiloxane (D5), and dodecamethylcyclohexasiloxane (D6) were identified. Depending on the siloxane species and type of tissue analyzed, siloxane levels in the range of about 10-1,400 ng g(-1) were detected; total silicon was found in all tissue samples in the range of about 8,900-85,000 ng g( 1). Higher platinum levels ranging from 25-90 ng g(-1 )were detected in fibrin layer and fat tissue of two patients with prostheses. No siloxanes were detected in control breast tissue samples. This investigation of human tissues by a combination of element-specific and species-specific analytical techniques clearly demonstrates for the first time that platinum and siloxanes leak from prostheses and accumulate in their surrounding tissues. PMID- 12589500 TI - Study of chromium-containing proteins in subcellular fractions of rat liver by enriched stable isotopic tracer technique and gel filtration chromatography. AB - Ten male Wistar rats were intravenously injected with a single approximately physiological dose of enriched stable isotopic Cr-50 tracer solution (200 ng (50)Cr(3+)/100 g body wt). The fundamental distribution patterns of the chromium containing proteins in the nucleic, mitochondrial, lysosomal, microsomal, and cytosolic subcellular fractions of the rat liver were investigated by means of Sephadex G-100 gel chromatography combined with neutron activation analysis via (50)Cr (n, gamma) (51)Cr reaction. In total, nine kinds of Cr-containing proteins were found in the five subcellular fractions, whose relative molecular masses were 96.6+/-6.2, 68.2+/-1.4, 57.9+/-4.7, 36.6+/-1.2, 24.2+/-1.8, 14.0+/-1.5, 8.8+/-0.6, 6.9+/-0.4, and 4.2+/-0.4 kDa. Approximately 64.5% of Cr proteins accumulated in the cytosolic fraction. The second enriched part was the nucleic fraction; about 12.2% Cr proteins were stored in this section. The 4.2-kDa molecular mass might contain the so-called low molecular weight chromium containing substance; however, in this research, it was only observed in the mitochondria, lysosome, and microsome. In the mitochondrial fraction, most of the Cr proteins were present as relatively low molecular weight substances: about 56% of chromium-containing proteins had molecular masses < or =6.9 kDa. Nevertheless, more than 69% of Cr-containing proteins were observed with molecular masses > or =57.9 kDa in the liver cytosolic fraction. PMID- 12589501 TI - Cathodic adsorptive stripping square-wave voltammetric determination of nifedipine drug in bulk, pharmaceutical formulation and human serum. AB - Nifedipine is a calcium-channel antagonist drug used in the management of angina pectoris and hypertension through inhibition of calcium influx. A fully validated sensitive cathodic adsorptive stripping square-wave voltammetry procedure was optimized for the determination of the drug at trace levels. The procedure was based on the reduction of the nitrophenyl group after the interfacial accumulation of the drug onto a hanging mercury drop electrode in Britton Robinson buffer of pH 11.0. The optimal conditions of the procedure were found to be: accumulation potential=-0.9 V vs. Ag/AgCl/KCl(s)), accumulation time=30 s, scan increment=10 mV, pulse amplitude=50 mV and frequency=120 Hz. Under these conditions, a well-defined peak was obtained; its peak current showed a linear dependence on drug concentration in the range of 2x10(-9)-2x10(-7) mol L(-1) bulk nifedipine. The mean recoveries based on eight replicate measurements for 1x10( 8) and 5x10(-8) mol L(-1) bulk nifedipine solutions were 98.46+/-0.86% and 98.23+/-0.92%, respectively. A detection limit of 3.42x10(-10) mol L(-1) bulk nifedipine was achieved. The procedure was successfully applied for assay of the drug in tablets and spiked human serum with mean recoveries of 101.95+/-1.42% and 98.70+/-0.63%, respectively. The limit of detection of the drug in spiked human serum was found to be 3.90x10(-10) mol L(-1). PMID- 12589502 TI - Determination of antioxidant activity of phenolic antioxidants in a Fenton-type reaction system by chemiluminescence assay. AB - The hydroxyl radical (*OH) has been implicated in various diseases, and it is therefore important to establish efficient methods to screen hydroxyl radical scavengers for antioxidant therapy. In this paper, a simple chemiluminescence assay was established to evaluate the *OH-scavenging capacity of phenolic compounds. This assay took advantage of the transient property of the Fenton reaction and the reaction between luminol and the hydroxyl radical, and effectively avoided the pro-oxidant action of some phenolic compounds. Fifteen phenolic compounds were assessed for their antioxidant activity in the Fenton reaction system, and even in the case of "pro-oxidants" that were excluded from the widely used deoxyribose (DR) assay. Since it overcomes the challenges that the traditional DR assay encounters, our method has promising applicative values: it is low-cost, time-saving, and reliable. It would also be more favorable than electron spin resonance (ESR) and radiolysis technology, which are known to be expensive and not commonly available to those specialized in free radical biology and medicine. PMID- 12589503 TI - Determination of nitrated polycyclic aromatic hydrocarbons in diesel particulate related standard reference materials by using gas chromatography/mass spectrometry with negative ion chemical ionization. AB - Gas chromatography/mass spectrometry (GC/MS) with negative ion chemical ionization (NICI) detection was utilized for quantitative determination of nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) in diesel particulate related standard reference materials (SRMs). Prior to GC/MS analysis, isolation of the nitro-PAHs from the complex diesel particulate extract was accomplished using solid phase extraction (SPE) and normal-phase liquid chromatographic (LC) fractionation using an amino/cyano stationary phase. Concentrations of eight to ten mononitro-PAHs and three dinitropyrenes were determined in three diesel particulate-related SRMs: SRM 1650a Diesel Particulate Matter, SRM 1975 Diesel Particulate Extract, and SRM 2975 Diesel Particulate Matter (Industrial Forklift). The results from GC/MS NICI using two different columns (5% phenyl methylpolysiloxane and 50% phenyl methylpolysiloxane) were compared to each other and to results from two other laboratories for selected nitro-PAHs. 1-Nitropyrene was the most abundant nitro-PAHs in each of the diesel particulate SRMs (19.8+/ 1.1 micro g g(-1) particle in SRM 1650a and 33.1+/-0.6 micro g g(-1) particle in SRM 2975). Three dinitropyrene isomers were measured in SRM 1975 at 0.5-1.4 micro g g(-1) extract and in SRM 2975 at 1-3 micro g g(-1) particle. PMID- 12589504 TI - Analysis of PAH compounds in soil with on-line coupled pressurised hot water extraction-microporous membrane liquid-liquid extraction-gas chromatography. AB - Pressurised hot water extraction (PHWE) was coupled on-line with microporous membrane liquid-liquid extraction (MMLLE) and gas chromatography (GC) in the analysis of polycyclic aromatic hydrocarbon (PAH) compounds in soil. The MMLLE serves as a trapping device after the PHWE. Water from PHWE is directed to the donor side of the membrane unit and the analytes are extracted to the acceptor solution on the other side of the membrane. The role of MMLLE is to clean and concentrate the extract, which is then transferred on-line to the GC via a sample loop and an on-column interface using partially concurrent solvent evaporation. Separate optimisation of MMLLE and simulations of the PHWE-MMLLE connection were carried out before the actual on-line coupling. After optimisation of the whole on-line system, the efficiencies of the PHWE-MMLLE-GC and PHWE-solid-phase trap extractions were compared. The PHWE-MMLLE-GC method allowed on-line analysis of soil samples. The method was linear, with limits of detection in the range 0.05 0.13 ng and limits of quantification 0.65-1.66 microg g(-1). Comparison of the results with those obtained by other techniques confirmed the good performance. PMID- 12589505 TI - Labile rhizosphere soil solution fraction for prediction of bioavailability of heavy metals and rare earth elements to plants. AB - A labile rhizosphere soil solution fraction has been recommended to predict the bioavailability of heavy metals and rare earth elements to plants. This method used moist rhizosphere soil in combination with a mixture of 0.01 mol L(-1) of low-molecular-weight organic acids (LMWOAs) as extractant. The extracted soil solutions were fractionated into two colloidal fractions of <0.45 microm (F(3)) and <0.2 microm (F(2)), and one truly dissolved fraction including free metal ions and inorganic and organic complexes (fractionr(0.2 microm, LMWOAs) approximately r(0.45 microm, LMWOAs). In the case of rare earth elements the good correlation was obtained for both the wheat roots and shoots. Generally, the correlation coefficients obtained by LMWAOs were better than that obtained by the first step of BCR method. Therefore, LMWAOs and F(lrss) were strongly recommended to predict the bioavailability of metals in soil pools to plants. PMID- 12589506 TI - Highly sensitive and selective spectrophotometric method for determination of trace gold in geological samples with 5-(2-hydroxy-5-nitrophenylazo)rhodanine. AB - A excellent sensitive and selective method for spectrophotometric determination of trace gold has been developed, the method is based on the color reaction of gold(III) with new reagent 5-(2-hydroxy-5-nitrophenylazo)rhodanine (HNAR). Under optimal conditions, HNAR reacts with gold(III) to form a 1:5 orange complex, which has an maximum absorption peak at 480 nm. Maximum enhancement of the absorbance of the complex was obtained in the presence of the mixed surfactant of Triton X-100 and CTMAB; the reaction completed rapidly and the absorbance is stable for 5 h at least at 20 degrees C; 0-48 microg L(-1) Au(III) obeyed Beer's law. The apparent molar absorptivity of the complex, Sandell's sensitivity, the limit of quantification, the limit of detection and relative standard deviation were found to be 2.0x10(6) L mol(-1) cm(-1), 0.000,098,483 micro g cm(-2), 1.02 ng mL(-1), 0.35 ng mL(-1) and 1.09%, respectively. The effect of co-existing ions was studied seriously; most metal ions can be tolerated in considerable amounts. Its sensitivity and selectivity are remarkably superior to other reagents in the literature. The proposed method was used successfully to determine trace gold in geological samples. Moreover, the synthesis, characteristics and analytical reaction of HNAR with gold are also described in detail. PMID- 12589507 TI - Applying non-parametric statistical methods to the classical measurements of inclusion complex binding constants. AB - A study on using non-parametric statistical methods was carried out to calculate the binding constant of an inclusion complex and to estimate its associated uncertainty. First, a correct evaluation of the stoichiometry was carried out in order to ensure an accurate determination of the binding constant. For this purpose, the modified Benesi-Hildelbrand method had been previously applied. Then, four statistical methods (three non-parametric methods: two bootstrap approaches, the jackknife method and a parametric one: Fieller's theorem) were employed in order to compute the binding constant. The results obtained from applying these methods and the combination of the methods: jackknife after bootstrap and bootstrap after jackknife were compared. The best results in terms of accuracy were obtained from the application of a bootstrap method: the resampling residuals approach. These procedures were applied to the inclusion complex 2-hydroxil-propyl-beta-cyclodextrin-2,4-dichloro-phenoxyacetic, which shows photochemically-induced fluorescence. PMID- 12589508 TI - Flow-injection chemiluminescence determination of chlorinated isocyanuric acids. AB - A rapid and sensitive flow-injection chemiluminescence method is described for the determination of dichloro- and trichloroisocyanuric acids based on the chemiluminescence produced during their reaction with luminol in alkaline medium. The effects of analytical and flow-injection variables on these chemiluminescence systems and determination of both oxidants are discussed. The optimized method yielded 3sigma detection limits of 8x10(-8) and 5x10(-8) mol L(-1) for the sodium dichloroisocyanurate and trichloroisocyanuric acid, respectively. The optimum conditions were found to be as follows: NaOH, 1x10(-1) mol L(-1); luminol, 5x10( 3) mol L(-1); KI, 2x10(-3) mol L(-1) and flow rate, 3.5 mL min(-1). PMID- 12589509 TI - Speciation of mercury in soil and sediment by selective solvent and acid extraction. AB - In order to characterize the mercury hazard in soil, a sequential extraction scheme has been developed to classify mercury species based on their environmental mobility and/or toxicity for either routine lab analysis or on-site screening purposes. The alkyl mercury species and soluble inorganic species that contribute to the major portion of potential mercury toxicity in the soil are extracted by an acidic ethanol solution (2% HCl+10% ethanol solution) from soil matrices as "mobile and toxic" species. A High-Performance Liquid Chromatography (HPLC) system coupled with Inductively Coupled Plasma Mass Spectrometry (ICP-MS) detection has been developed to further resolve the species information into soluble inorganic species (Hg(2+)), methylmercury(II) (MeHg(+)) and ethylmercury(II) (EtHg(+)) species. Alternatively, these species can be separated into "soluble inorganic mercury" and "alkyl mercury" sub-categories by Solid Phase Extraction (SPE). A custom Sulfydryl Cotton Fiber (SCF) material is used as the solid phase medium. Optimization of the SCF SPE technique is discussed. Combined with a direct mercury analyzer (DMA-80), the SCF SPE technique is a promising candidate for on-site screening purposes. Following the ethanol extraction, the inorganic mercury species remaining in soil are further divided into "semi-mobile" and "non-mobile" sub-categories by sequential acid extractions. The "semi-mobile" mercury species include mainly elemental mercury (Hg) and mercury-metal amalgams. The non-mobile mercury species mainly include mercuric sulfide (HgS) and mercurous chloride (Hg(2)Cl(2)). PMID- 12589510 TI - Superheated water extraction of cholesterol from solid food. AB - A method based on superheated water extraction has been developed for the removal of cholesterol from solid food thus providing a clean approach by avoiding the use of organic solvents. A preconcentration step was also studied with the aim of solving the problem derived from low-cholesterol-content samples and dilution effect. The research also involved the optimisation of the parameters affecting the extraction process by a central composite experimental design as well as a univariate study of the optimisation of the preconcentration step. The time required for total removal of the target compound was 60 min. The method was validated using a certified reference material (NIST-CRM 1845) and was used to analyse food samples within a wide range of cholesterol concentrations. The efficiency for the CRM was 105%. The precision of the method yielded values less than 6.5% (expressed as relative standard deviation) in all instances. PMID- 12589511 TI - Use of factorial design and Doehlert matrix for multivariate optimisation of an on-line preconcentration system for lead determination by flame atomic absorption spectrometry. AB - A system for on-line preconcentration and determination of lead by flame atomic absorption spectrometry (FAAS) was proposed. It was based on the sorption of lead(II) ions on a minicolumn of polyurethane foam loaded with 2-(2-thiazolylazo) 5-dimethylaminophenol (TAM). The optimisation step was carried out using two level full factorial and Doehlert designs for the determination of the optimum conditions for lead preconcentration. The proposed procedure allowed the determination of lead with a detection limit of 2.2 microg L(-1), and a precision, calculated as relative standard deviation (RSD), of 2.4 and 6.8 for a lead concentration of 50.0 and 10.0 microg L(-1), respectively. A preconcentration factor of 45 and a sampling frequency of 27 samples per hour were obtained. The recovery achieved for lead determination in the presence of several cations demonstrated that this procedure has enough selectivity for analysis of environmental samples. The validation was carried out by analysis of certified reference material. This procedure was applied to lead determination in natural food. PMID- 12589512 TI - Sorption of trace metals on human hair and application for cadmium and lead pre concentration with flame atomic absorption determination. AB - Human hair shavings were characterized as a sorbent for trace metals. At pH 7.0 metal sorption follows the order Pb(II)>Cd(II)>Cr(VI)>Fe(III)>Cu(II)>Ni(II)>Mn(VI). Metal recovery is quantitative for Pb and Cd after 30 min of equilibration. Recovery of other metals is less quantitative and varies with pH. For example, while Cu is best recovered at pH 5, Ni and Mn are sorbed optimally in the basic pH region. Sorbed metals can be washed off the sorbent with 0.5 mol L(-1) strong mineral acids or more completely with 0.1 mol L(-1) ethylenediaminetetraacetic acid (EDTA). Typical sorption isotherms were obtained for Cd and Pb with sorption capacities of 39 and 26 micromol g(-1), respectively. Hair sorbent was used for 40-fold pre-concentration of Cd and Pb from treated wastewater samples followed by flame atomic absorption spectroscopic (FAAS) determination. Comparison of the data obtained for lead and cadmium by the proposed pre-concentration method with that by graphite furnace atomic absorption spectroscopy (GFAAS) showed 79 to 86% recovery and comparable analytical precision. Common cations and anions at the levels normally present in natural water do not interfere in the proposed pre-concentration-FAAS method. PMID- 12589513 TI - Condenser-type diffusion denuders for the collection of sulfur dioxide in a cleanroom. AB - High-efficiency condenser-type diffusion denuders of cylindrical and planar geometries are described. The film condensation of water vapor onto a cooled denuder surface can be used as a method for collecting water-soluble gases. By using SO(2) as the test gas, the planar design offers quantitative collection efficiency at air sampling rates up to 5 L min(-1). Coupled to ion chromatography, the limit of detection (LOD) for SO(2) is 0.014 ppbv with a 30 min successive analysis sequence. The method has been successfully applied to the analysis of temperature- and humidity-controlled cleanroom air. PMID- 12589514 TI - Solid-phase micro extraction (SPME) and headspace derivatization of clenbuterol followed by GC-FID and GC-SIMMS quantification. AB - Solid-phase micro extraction (SPME) and on-fiber derivatization followed by Gas Chromatography coupled with Flame Ionization Detection (GC-FID) or Selected Ion Monitoring Mass Spectrometry (GC-SIMMS) allows for simple yet sensitive quantification for the hexamethyldisilazane derivative of the beta-agonist clenbuterol. Using an 85- micro m polyacrylate fiber, the analysis method is optimized with respect to extraction time, derivatization time and temperature, and solution pH. In addition, the use of a rapid temperature ramping injection port allows for optimization of fiber desorption conditions. Under optimal conditions, the limits of detection for the hexamethyldisilazane derivative of clenbuterol are 1.1 ppb by FID and 0.20 ppb by SIMMS. PMID- 12589515 TI - Investigation of antioxidant and catalytic properties of some biologically active substances by voltammetry. AB - Antioxidants play a major role in protecting biological systems against many incurable diseases. The biological activity of 12 plant aqueous-alcohol extracts, some standard antioxidants (vitamin C, glucose, resorcinol, and catechol), Na(2)SO(3), humic acids, phthalocyanines, and chlorophyll have been investigated in this work together with evaluation of their influence on the kinetics of the oxygen electroreduction. Finally the use of these substances for prophylactic purposes has been recommended. PMID- 12589516 TI - Occupancy of 5-HT1A receptors by clozapine in the primate brain: a PET study. AB - RATIONALE: The pharmacological mechanism underlying the atypical properties of the antipsychotic drug clozapine remains to be identified. The serotonin 5 hydroxytryptamine-1A (5-HT(1A)) receptor subtype has been suggested to play a role in the pathophysiology of schizophrenia and is one among several central neuroreceptors for which clozapine has moderate affinity in vitro. OBJECTIVE: The aim of this positron emission tomography (PET) study was to determine 5-HT(1A) receptor occupancy in the monkey brain after IV injection of clozapine in doses that previously have been shown to give plasma concentrations representative of the 200 to 800 mg oral dose range recommended for clinical management of patients. METHODS: Each of four cynomolgus monkeys was examined three times on the same day with PET and the radioligand [carbonyl-(11)C]WAY-100635. The first measurement was performed at baseline conditions, the second after clozapine 1.5 mg/kg and the third after 6 mg/kg. Two additional monkeys were examined at baseline and after 15 mg/kg IV. Central 5-HT(1A) receptor occupancy was calculated using an equilibrium-ratio analysis. RESULTS: The occupancy ranged from 23 to 34% after 1.5 mg/kg clozapine and from 36 to 49% after 6 mg/kg in different brain regions of the four monkeys. The regional receptor occupancy values after 15 mg/kg were between 39 and 51% in the two monkeys. There was no evident difference between the frontal cortex, the temporal cortex and the raphe nucleus. CONCLUSION: The study shows that clozapine occupies 5-HT(1A) receptors in the primate brain at clinically representative plasma concentrations. The results support that the 5-HT(1A) receptor is a candidate target for the atypical drug actions of clozapine. PMID- 12589517 TI - Age differences in the sensitivity to clomipramine in an animal model of obsessive-compulsive disorder. AB - RATIONALE: Subtypes of obsessive-compulsive disorder (OCD) related to age could determine differential response to treatment. OBJECTIVES: To explore possible age differences in the effect of clomipramine in an animal model of OCD. METHODS: The deficits on spontaneous alternation produced by 8-OH-DPAT and the preventing actions of clomipramine, desipramine and WAY 100635 were compared between young and adult rats. RESULTS: No age differences were found in spontaneous alternation. The 5-HT(1A) agonist, 8-OH-DPAT (0.031, 0.125, 0.5 and 2.0 mg/kg, 15 min) produced perseveration in young and adult rats. However, young rats were sensitive to a lower dose of 8-OH-DPAT. Clomipramine (10 mg/kg per three administrations) completely prevented the action of 8-OH-DPAT (0.5 mg/kg) in adult rats. However, this treatment as well as higher doses (15 mg/kg 3 administrations) or injected for longer periods (10 mg/kg 5 administrations) produced weak protective effects (versus 0.125 mg/kg 8-OH-DPAT) or had no action (versus 0.5 mg/kg 8-OH-DPAT) in young animals. WAY 100 635 (0.5 mg/kg) blocked the action of 8-OH-DPAT (0.5 mg/kg) in both young and adult rats. Desipramine (10 mg/kg/3 administrations) lacked of a preventive effect on the 8-OH-DPAT (0.5 mg/kg) action. This result indicated that the 5-HT(1A) receptor is involved in the deficits on spontaneous alternation produced by 8-OH-DPAT. CONCLUSIONS: The present data shows important age differences in the effect of clomipramine in a model of OCD. Such differences could be relevant for the age variations in the response to treatment in clinical practice. PMID- 12589518 TI - Relationships between individual sensitivity to CS- and cocaine-induced reinstatement in the rat. AB - RATIONALE: Maintaining abstinence is highly challenging for cocaine ex-users. Exposure to drug conditioned stimuli (CS) and to low doses of cocaine can provoke craving in humans and reinstate self-administration (SA) behavior in animal models. Whether drug- and CS-induced reinstatement depend on the same biological substrates remains controversial. OBJECTIVES: We investigated the relationships between cocaine- and CS-induced SA reinstatement within the same individuals as a function of the duration of the withdrawal period after cessation of extended cocaine SA. METHODS: Sprague-Dawley rats were trained for cocaine intravenous SA (0.8 mg/kg per infusion) during 74 sessions (2 h daily exposure to cocaine) and submitted to withdrawal. Five and 30 days after the end of SA, cocaine- and CS induced reinstatement were tested. RESULTS: Both after a short and a long withdrawal, CS- and cocaine-induced reinstatement were not related. Furthermore, cocaine-induced reinstatement measured after a short and a long withdrawal was positively related while CS-induced reinstatement was not. The sensitivity of an individual to cocaine-induced reinstatement is not related to its sensitivity to CS-induced reinstatement. Furthermore, vulnerability to cocaine-induced reinstatement is determined quickly after SA cessation and is a long lasting state, whilst vulnerability to CS-induced reinstatement develops quickly or slowly depending on the individual. CONCLUSIONS: These results support the view that cocaine and CS induce reinstatement through different mechanisms. They imply that reinstatement in drug abuse is a heterogeneous condition with some individuals being more sensitive to one factor than to the other. Research for effective anti-relapse therapies should take these elements into account. PMID- 12589519 TI - The effect of neurotoxic doses of methamphetamine on methamphetamine-conditioned place preference in rats. AB - RATIONALE: Methamphetamine has been shown to produce neurotoxicity demonstrated by depletions of dopamine and serotonin in the striatum and nucleus accumbens. OBJECTIVE: The current study examined the effects of neurotoxic doses of methamphetamine on the rewarding effect of subsequent administration of methamphetamine assessed by the conditioned place preference (CPP) procedure. METHODS: Male and female rats were treated with a neurotoxic regimen of methamphetamine (4 x 10 mg/kg, s.c., once every 2 h) or saline, and concentrations of dopamine, 3,4-dihydroxyphenylacetic acid, serotonin, and 5 hydroxyindoleacetic acid were measured 15 days later in the striatum, nucleus accumbens, and prefrontal cortex (PFC). In another experiment, male rats were given methamphetamine neurotoxic treatment or saline and were then conditioned 7 days later with methamphetamine (0.1, 0.3, or 1.0 mg/kg, s.c.) or saline using a four-trial CPP procedure. Locomotor activity was also measured during the conditioning sessions to investigate whether or not the neurotoxic methamphetamine treatment altered locomotor activity following a subsequent methamphetamine challenge. RESULTS: Males and females did not differ significantly in the amount of neurochemical depletion produced by methamphetamine in any brain region. Collapsed across sex, dopamine was significantly depleted in nucleus accumbens (25%) and striatum (51%); serotonin was significantly depleted in nucleus accumbens (35%), striatum (34%) and PFC (33%). The methamphetamine challenge dose dependently increased locomotor activity, but the increase was not affected by treatment with neurotoxic doses of methamphetamine. In contrast, treatment with neurotoxic doses of methamphetamine enhanced CPP at the intermediate conditioning dose (0.3 mg/kg). CONCLUSIONS: These results indicate that the rewarding effect of methamphetamine is enhanced by prior treatment with neurotoxic doses of methamphetamine, suggesting either a compensatory hyperfunctioning of spared dopamine neurons or a loss of inhibitory control from serotonergic input. PMID- 12589520 TI - Assessment of a combination of physostigmine and scopolamine as pretreatment against the behavioural effects of organophosphates in the common marmoset (Callithrix jacchus). AB - RATIONALE: There is a requirement to ensure that UK armed forces are provided with the best possible medical countermeasures to prevent or mitigate the effects of exposure to nerve agents. When pretreatments are under consideration, it is of particular importance to ensure that they do not in themselves give rise to adverse effects and do not exacerbate the effects of agent exposure. OBJECTIVES: The present study was designed to address these considerations for a combination of physostigmine and scopolamine as a potential pretreatment regimen. METHODS: Common marmosets were trained to perform a two-choice discrimination serial reversal task, and baseline data were collected. Subjects received a dose of either soman or sarin after 2 weeks of pretreatment with either saline or physostigmine and scopolamine via miniosmotic pump. RESULTS: No effects of physostigmine and scopolamine were seen on task accuracy or response rates. Neither accuracy of reversal performance nor number of responses made were significantly changed by administration of either soman or sarin subsequent to pretreatment with physostigmine/scopolamine. In the groups pretreated with saline, performance of the behavioural task, in terms of responses made, was virtually abolished on the day the OP was administered, but a significant increase in accuracy of performance was seen over the 2- to 14-day period following administration. CONCLUSIONS: A combination of physostigmine and scopolamine, which is known to protect against nerve-agent lethality, offers protection against the effects of soman and sarin on behavioural performance, as measured by a discrimination reversal task. The improved performance observed following nerve agent requires further investigation. PMID- 12589521 TI - EEG effects of buspirone and pindolol: a method of examining 5-HT1A receptor function in humans. AB - RATIONALE: An involvement of 5-HT(1A) receptors is postulated in the pathophysiology of affective disorders and mechanism of action of antidepressants. Methods for studying their functional integrity in humans are, however, limited. Preliminary data suggests that activation of somatodendritic 5 HT(1A) receptors cause a negative shift in the EEG frequency spectrum. Animal research suggests that pindolol is an agonist at these receptors but an antagonist at postsynaptic 5-HT(1A) receptors. OBJECTIVE: We postulated that while pindolol would antagonise known postsynaptic mediated neuroendocrine responses to the 5-HT(1A) agonist buspirone, both drugs would have a similar effect on the EEG frequency spectrum. METHODS: Fourteen healthy men were administered placebo or pindolol (20 mg orally) 90 min before placebo or buspirone (30 mg orally) in a double blind cross-over study. Plasma prolactin and growth hormone were assayed and EEGs recorded before and after drug administration. RESULTS: A significant negative shift in the EEG frequency spectrum was found for both buspirone and pindolol, with the combination producing a similar effect to each drug alone. In contrast, the neuroendocrine response to buspirone was significantly attenuated by pindolol. CONCLUSIONS: The data obtained are consistent with the EEG effects of buspirone and pindolol being mediated by somatodendritic 5-HT(1A) receptors, in contrast to the neuroendocrine response, which is known to be mediated by postsynaptic receptors. The development of this novel method of assessing somatodendritic 5-HT(1A) receptors in humans is a potentially important advance which may allow the testing of hypotheses of its involvement in depression and response to antidepressants. PMID- 12589522 TI - Cardiovascular effects of methylphenidate in humans are associated with increases of dopamine in brain and of epinephrine in plasma. AB - RATIONALE: The cardiovascular effects of psychostimulant drugs (methylphenidate, amphetamine, cocaine) have been mostly associated with their noradrenergic effects. However, there is some evidence that dopaminergic effects are involved in the cardiovascular actions of these drugs. Here, we evaluated this association in humans. METHODS: Positron emission tomography (PET) and [(11)C]raclopride, a dopamine (DA) D2 receptor radioligand that competes with endogenous DA for occupancy of the D2 receptors, were used to measure changes in brain DA after different doses of intravenous methylphenidate in 14 healthy subjects. Cardiovascular (heart rate and blood pressure) and catecholamine (plasma epinephrine and norepineprhine) responses were determined in parallel to assess their relationships to methylphenidate-induced changes in brain DA. RESULTS: Methylphenidate administration significantly increased heart rate, systolic and diastolic blood pressures and epinephrine concentration in plasma. The increases in blood pressure were significantly correlated with methylphenidate-induced increases of DA in striatum (r>0.78, P<0.001) and of plasma epinephrine levels (r>0.82, P<0.0005). In turn methylphenidate-induced DA increases in striatum were correlated with increases of epinephrine in plasma (r=0.85, P<0.0001). Subjects in whom methylphenidate did not increase DA had no change in blood pressure or in plasma epinephrine concentration. DISCUSSION: These results are consistent with the hypothesis that methylphenidate-induced increases in blood pressure are in part due to its central dopaminergic effects. They also suggest that methylphenidate's pressor effects may be in part mediated by DA-induced increases in peripheral epinephrine. PMID- 12589523 TI - Suppression of cocaine- and food-maintained behavior by the D2-like receptor partial agonist terguride in squirrel monkeys. AB - RATIONALE: The D2-like receptor partial agonist terguride has a profile of behavioral effects in rats that suggests potential benefit as a pharmacotherapy for cocaine addiction. OBJECTIVES: The present study investigated the effects of terguride on cocaine- and food-maintained behavior in squirrel monkeys. METHODS: Squirrel monkeys were trained to respond on a second-order schedule (FI 10 min, FR 10 or 30:S) of either i.v. cocaine injection or food pellet delivery. Additional monkeys were studied using quantitative observational techniques to construct side effect profiles. Under each procedure, the effects of terguride were compared with those of the reference D2-like receptor antagonist nemonapride and the D2-like receptor full agonist quinpirole. RESULTS: Terguride and nemonapride, but not quinpirole, dose-dependently reduced cocaine-maintained behavior. In animals self-administering food, terguride decreased response rates at doses lower than those required to suppress cocaine-maintained behavior. Effective doses of terguride had no systematic effect on motor activity or muscle rigidity, whereas effective doses of nemonapride virtually eliminated motor activity and induced severe catalepsy. The primary observable effects of terguride were a modest increase in self-directed behavior (a D2-receptor agonist like effect) at intermediate doses and a small increase in static posture (a D2 receptor antagonist-like effect) at the highest dose tested. CONCLUSIONS: The results suggest that terguride has advantages over conventional D2-like receptor antagonists and agonists as a candidate pharmacotherapy for cocaine abuse; however, terguride significantly reduces food-maintained behavior at lower doses than those needed to decrease cocaine-maintained behavior suggesting limitations on the utility of terguride as a medication for cocaine addiction. PMID- 12589524 TI - Differences in platelet serotonin transporter sites between African-American tobacco smokers and non-smokers. AB - RATIONALE: The serotonin transporter (5HTT) regulates the magnitude and duration of serotonergic neurotransmission. Although nicotine and other constituents of tobacco smoke may influence serotonin turnover among animals, few studies have examined whether smoking is associated with alteration in 5HTT in humans. OBJECTIVE: We investigated whether tobacco smokers and non-smokers differed in platelet tritiated paroxetine binding, a measure of 5HTT sites, and whether severity of nicotine dependence (ND) was related to 5HTT measures. METHODS: Tritiated paroxetine binding sites on platelets were assayed in 26 African American smokers and 30 non-smokers. Severity of smoking was assessed using the Fagerstrom Test for Nicotine Dependence (FTND). Relationships between FTND scores and maximum number of transporter sites (B(max)) and affinity constant (K(d)) of paroxetine binding were determined. RESULTS: B(max) values showed a significant negative correlation with FTND scores (rho=-0.28, P<0.01). Notably, smokers with higher ND had significantly lower B(max) compared to those with lower ND and non smokers; the latter two groups did not differ in B(max) ( F=3.92, P<0.05). Smokers scored higher on impulsivity than non-smokers, however, behavioral variables did not influence the relationship of smoking with B(max). Age, gender and K(d) values were not associated with smoking or B(max). CONCLUSIONS: Smoking, in particular higher nicotine dependence, appears to be correlated with decreased density of platelet 5HTT sites in African-Americans. The nature of the relationship and whether similar changes occur in the brain merit further investigation. PMID- 12589525 TI - Caffeine reinforces flavour preference in caffeine-dependent, but not long-term withdrawn, caffeine consumers. AB - RATIONALE: Previous studies have shown that caffeine can reinforce flavour liking in overnight deprived moderate caffeine consumers (e.g. average of 250 mg/day) but not in low consumers (<120 mg/day). However, it is not possible to determine whether the difference between moderate and low caffeine consumers results from pre-existing individual differences in response to caffeine, or results directly from the different amounts of caffeine they habitually consume. If the former were true, then moderate consumers who are completely withdrawn should still manifest the flavour conditioning effect. Conversely, if the latter were true, consumers who are completely withdrawn should not manifest the effect. OBJECTIVES: To examine whether moderate caffeine consumers who have been fully withdrawn from caffeine manifest the flavour conditioning effect. METHODS: In a double-blind study, 48 moderate caffeine consumers refrained from consuming caffeine for 4 weeks and were given replacement drinks to consume, which were either caffeinated (maintained group) or decaffeinated (withdrawn group). In the final 2 weeks, all subjects evaluated a novel drink containing either 100 mg caffeine or placebo on four non-consecutive days. RESULTS. The rated pleasantness of the novel drink containing caffeine increased over the four test days in the group maintained on caffeine, but pleasantness of the same drink fell significantly in the withdrawn group. CONCLUSIONS: These data suggest that the ability of caffeine to reinforce changes in flavour liking are driven by the alleviation of withdrawal symptoms among habitual caffeine consumers and provide further support for the negative reinforcement theory. PMID- 12589526 TI - Chronic treatment with caffeine blunts the hyperlocomotor but not cognitive effects of the N-methyl-D-aspartate receptor antagonist MK-801 in mice. AB - RATIONALE: Administration of N-methyl- d-aspartate (NMDA) receptor antagonists produce hyperlocomotion and cognitive deficits in rodents. Activation of NMDA receptors promotes adenosine release, and adenosine agonists prevent central effects of NMDA receptor antagonists. We hypothesized that if NMDA receptor antagonists require adenosine to produce behavioral effects, mice tolerant to the adenosine receptor antagonist caffeine would have a diminished response to NMDA receptor antagonists. OBJECTIVES: To evaluate MK-801-induced hyperlocomotion and cognitive deficits after chronic caffeine treatment in mice. METHODS: Locomotor activity was analyzed in a computerized system, spontaneous alternation was assessed in the Y-maze and long-term memory was assessed with the inhibitory avoidance task in mice. RESULTS: Mice chronically treated with caffeine in drinking solution (1 mg/ml for 7 days) presented normal habituation and substantial tolerance to acute caffeine (30 mg/kg, i.p.) locomotor effects. MK 801 (0.25 mg/kg, i.p.) produced pronounced hyperlocomotion in water-treated mice, but this effect was abolished in caffeine-drinking mice. Chronic caffeine treatment had no influence on either normal or MK-801-induced deficits in spontaneous alternation and inhibitory avoidance tasks. CONCLUSION: Hyperlocomotion induced by MK-801 may be mediated by reduced adenosinergic activity. These results also suggest that locomotor and cognitive effects of MK 801 can be dissociated and are distinctly modulated. Finally, these findings agree with the adenosine hypofunction model of schizophrenia, since NMDA receptor antagonists are a pharmacological model for this disorder. PMID- 12589527 TI - Antagonism of nitrous oxide-induced anxiolytic-like behavior in the mouse light/dark exploration procedure by pharmacologic disruption of endogenous nitric oxide function. AB - RATIONALE: Previous studies have shown the anxiolytic-like effects of nitrous oxide (N(2)O) to be sensitive to antagonism by non-specific inhibitors of nitric oxide synthase (NOS). OBJECTIVES: The present study was conducted to demonstrate further the involvement of nitric oxide (NO) and ascertain whether a specific isoform of NOS is involved in N(2)O-induced behavior in mice. METHODS: Male NIH Swiss mice were tested in the light/dark exploration test to determine how N(2)O induced behavior was affected by the following pretreatments: the NO scavenger hemoglobin (Hb); the selective nNOS-inhibitor S-methyl- l-thiocitrulline (SMTC); the selective eNOS-inhibitor N(5)-(1-iminoethyl)- l-ornithine ( l-NIO); and the selective iNOS-inhibitor 2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine (AMT). Furthermore, NOS activity was assessed in the whole brain as well as five brain areas of N(2)O- versus room air-exposed mice to determine the effects of N(2)O on NOS activity. RESULTS: The behavioral effects of N(2)O in the light/dark exploration test were significantly attenuated following pretreatment with Hb (2.0 nmol, i.c.v.), SMTC (0.3 micro g and 1.0 micro g per mouse, i.c.v.) and the higher dose of l-NIO (30 mg/kg, s.c.). However, the N(2)O-induced behavioral effect was unaltered by pretreatment with either the lower dose of l-NIO (10 mg/kg, s.c.) or AMT (1.0 mg/kg and 3.0 mg/kg, s.c.). Finally exposure to 50% N(2)O for 15 min significantly increased NOS activity in the cerebellum and corpus striatum but not in other brain regions or whole brain. CONCLUSION: These findings provide further support for the hypothesis that NO is involved in N(2)O induced anxiolytic-like behavior and that this NO is the product of nNOS enzyme activity. PMID- 12589528 TI - Cardiac troponin I in patients with severe exacerbation of chronic obstructive pulmonary disease. AB - OBJECTIVES: Co-morbid conditions including risk factors for cardiovascular diseases and left ventricular dysfunction are common in patients with chronic obstructive pulmonary disease (COPD). This study assessed the incidence of cardiac troponin I (cTnI) elevation, a specific marker for cardiac injury, and its prognostic significance during severe exacerbation of COPD. DESIGN: Prospective cohort study. SETTING: Two intensive care units. PARTICIPANTS: Seventy-one consecutive patients admitted for severe exacerbation of COPD. INTERVENTION: None. MEASUREMENTS AND RESULTS: Cardiac troponin I was assayed in blood samples obtained on admission and 24 h later (Stratus II immunoassay analyser, Dade International). Levels above 0.5 ng/ml were considered positive. The following data were recorded prospectively: clinical symptoms, co morbidities, cause of the exacerbation, diagnostic procedures and treatment, general severity score (SAPS II) and in-hospital outcome. CTnI was positive in 18% of patients (95% confidence interval (CI(95)), 11-29%), with a median value at 1.00 ng/ml; CI(95 )(0.60-1.70). Eighteen patients died in the hospital (25%; CI(95), 17-37%). Only cTnI (adjusted odds ratio (ORa), 6.52; CI(95),1.23-34.47) and SAPS II 24 h after admission (ORa, 1.07; CI(95), 1.01-1.13) were independent predictors of in-hospital mortality. CONCLUSION: Elevated cTnI is a strong and independent predictor of in-hospital death in patients admitted for acutely exacerbated COPD. PMID- 12589529 TI - Treatment with bovine surfactant in severe acute respiratory distress syndrome in children: a randomized multicenter study. AB - OBJECTIVE: To determine whether bovine surfactant given in cases of severe pediatric acute respiratory distress syndrome (ARDS) improves oxygenation. DESIGN: Single-center study with 19 patients, followed by a multicenter randomized comparison of surfactant with a standardized treatment algorithm. Primary endpoint PaO(2)/FIO(2) at 48 h, secondary endpoints: PaO(2)/FIO(2) at 2, 4, 12, and 24 h, survival, survival without rescue, days on ventilator, subgroups analyzed by analysis of variance to identify patients who might benefit from surfactant. SETTING: Multicenter study in 19 reference centers for ARDS. PATIENTS: Children after the 44th postconceptional week and under 14 years old, admitted for at least 4 h, ventilated for 12-120 h, and without heart failure or chronic lung disease. In the multicenter study 35 patients were recruited; 20 were randomized to the surfactant group and 15 to the nonsurfactant group. Decreasing recruitment of patients led to a preliminary end of this study. INTERVENTIONS: Administration of 100 mg/kg bovine surfactant intratracheally under continuous ventilation and PEEP, as soon as the PaO(2)/FIO(2) ratio dropped to less than 100 for 2 h (in the pilot study increments of 50 mg/kg as long as the PaO(2)/FIO(2) did not increase by 20%). A second equivalent dose within 48 h was permitted. RESULTS: In the pilot study the PaO(2)/FIO(2) increased by a mean of 100 at 48 h (n=19). A higher PaO(2)/FIO(2) ratio was observed in the surfactant group 2 h after the first dose (58 from baseline vs. 9), at 48 h there was a trend towards a higher ratio (38 from baseline vs. 22). The rate of rescue therapy was significantly lower in the surfactant group. Outcome criteria were not affected by a second surfactant dose (n=11). A significant difference in PaO(2)/FIO(2) in favor of surfactant at 48 h was found in the subgroup with an initial PaO(2)/FIO(2) ratio higher than 65 and in patients without pneumonia. CONCLUSIONS. Surfactant therapy in severe ARDS improves oxygenation immediately after administration. This improvement is sustained only in the subgroup of patients without pneumonia and that with an initial PaO(2)/FIO(2) ratio higher than 65 PMID- 12589530 TI - Effectiveness of non-invasive positive pressure ventilation differs between decompensated chronic restrictive and obstructive pulmonary disease patients. AB - OBJECTIVE: To compare the efficiency of non-invasive positive pressure ventilation (NPPV) in decompensated patients with either chronic obstructive pulmonary disease (COPD) or chronic restrictive pulmonary disease. DESIGN: Retrospective study. SETTING: A 17-bed intensive care unit in a university teaching hospital. SETTING: Sixty-four patients with COPD (age: 70+/-13 years, sex ratio: 37 male to 27 female patients, forced expiratory volume in 1 s: 31+/ 13% predicted) and 20 patients with chronic restrictive pulmonary disease (age: 75+/-9 years, sex ratio: 9 male to 11 female patients, total lung capacity: 57+/ 17% predicted) consecutively treated with NPPV (facial mask, pressure support ventilation (PSV) +/- PEEP) for acute respiratory failure. MEASUREMENTS AND RESULTS: There were no statistically significant differences between COPD and patients with chronic restrictive pulmonary disease in terms of cause of exacerbation, use of oxygen therapy or NPPV at home, severity of acute respiratory failure (ARF), mean delay from intensive care admission to initiation of NPPV and total duration of NPPV. Patients with chronic restrictive pulmonary disease had a lower success rate on NPPV (without need of tracheal intubation) than COPD (35% vs 67%, p=0.01). Causes of NPPV failure were not different between COPD and patients with restrictive disease. After 12 h of NPPV, restrictive patients who succeeded with NPPV had similar respiratory rate, minute ventilation and arterial blood gas to COPD patients. At the 3rd and 12th h of NPPV, improvements in pH and PaCO(2) were predictive of NPPV success in COPD, but not in restrictive patients. CONCLUSION: The results of this retrospective study suggest that the effectiveness of NPPV for acute decompensation is less in patients with chronic restrictive pulmonary disease as compared to COPD. PMID- 12589531 TI - Safety of percutaneous dilational tracheostomy in patients ventilated with high positive end-expiratory pressure (PEEP). AB - OBJECTIVE: To assess the impact of bronchoscopically guided percutaneous dilational tracheostomy (PDT) on oxygenation in patients with severe respiratory failure ventilated with high positive end-expiratory pressure (PEEP). DESIGN: Prospective clinical study. SETTING: Anaesthesiological ICU, referral centre for acute respiratory distress syndrome (ARDS) therapy, university hospital. PATIENTS: Mechanically ventilated patients with indication for PDT. Two hundred three consecutive PDTs were performed in 198 patients on either high (>10 mbar, n=88) or low (45 mmHg) in patients with neuromuscular disorders. SETTING: A rehabilitation hospital. PATIENTS: The study was performed in 95 neuromuscular patients; 52 were tracheostomized with a cuffless tracheostomy tube (invasive ventilation), and 43 received noninvasive ventilation. MEASUREMENTS AND RESULTS: The volume of air leaked (VL) and arterial carbon dioxide (PaCO(2)) were routinely measured during mechanical ventilation; PaCO(2) was also measured during spontaneous breathing. VL, expressed as a percentage of tidal volume, was higher in the hypercapnic group (32+/-14%, n=20) than the nonhypercapnic group ( vs. 20+/-14%). PaCO(2) during mechanical ventilation was correlated with both VL and the duration of ventilatory support per day; PaCO(2) during spontaneous breathing was correlated only with the volume of air leaked. In stepwise multiple regression analysis, air leaks contributed to 8% of the variance in PaCO(2) during mechanical ventilation, and daily duration of ventilatory support contributed 5%. In addition, reduction in VL with normalization of PaCO(2) was achieved in five of the noninvasively ventilated patients with persistent hypercapnia by using a chin strap. CONCLUSIONS: Air leaks during wakefulness are an important cause of persistent hypercapnia in both invasively and noninvasively ventilated neuromuscular patients. However, simple practical measures to reduce the volume of air leaks improve the efficacy of ventilation in these patients. PMID- 12589534 TI - Acinetobacter baumannii: a threat for the ICU? PMID- 12589535 TI - CC-chemokine activation in acute pancreatitis: enhanced release of monocyte chemoattractant protein-1 in patients with local and systemic complications. AB - OBJECTIVE: Systemic leukocyte activation is claimed to trigger inflammatory response and remote organ dysfunction in acute pancreatitis. Chemokines are inflammatory mediators with potent leukocyte-activating properties and have been shown to be involved in the pathophysiological process of experimental acute pancreatitis. However, as little is known about their role in human disease we investigated local and systemic concentrations of different CC-chemokine members in patients with acute pancreatitis. PATIENTS AND METHODS: We included 68 patients with acute pancreatitis in the present study. Local complications in terms of necrosis were present in 37 (54%) patients of whom 21 (57%) developed pancreatic infections. Monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1alpha (MIP-1alpha) and macrophage inflammatory protein 1beta (MIP-1beta) concentrations were measured daily over 2 weeks after study inclusion by ELISA in sera and lesser sac aspirates. RESULTS: MCP-1 serum concentrations showed a dramatic increase in patients who developed local complications and/or remote organ failure. Herein, a close correlation was found between the severity of remote organ failure and the degree of MCP-1 elevation. Multiple regression analysis identified pancreatic infections as well as renal and cardiocirculatory failure as independent variables associated with enhanced systemic MCP-1 release. MIP-1alpha levels remained unaffected by local complications and showed a significant increase only; if multiple organ dysfunction syndrome (MODS) developed or patients subsequently died. In contrast, MIP-1beta concentrations correlated with neither the presence nor the severity of any complication. Compared with systemic concentrations, local lesser sac aspirates revealed significantly higher levels of MCP-1, whereas MIP-1alpha and MIP-1beta were not different. CONCLUSIONS: Complicated acute pancreatitis is associated with significantly elevated local and systemic concentrations of the CC-chemokine MCP-1. Our results suggest that, among the CC-chemokine members investigated, MCP-1 might play a pivotal role in the pathological mechanism of complicated acute pancreatitis. PMID- 12589536 TI - Effects of non-invasive ventilation on middle ear function in healthy volunteers. AB - OBJECTIVES: To evaluate the effects of non-invasive ventilation (NIV) with facial mask or helmet on middle ear (ME). DESIGN. Prospective, randomised study. SETTING: University hospital. PARTICIPANTS: Ten healthy subjects randomly allocated in two groups of five subjects each. INTERVENTIONS: NIV for 1 h, with helmet (group H) or facial mask (group M). Flow-triggered pressure support was 10 cmH(2)O, PEEP 5 cmH(2)O, FiO(2) 0.21. MEASUREMENTS AND RESULTS: Impedenzometry was performed before NIV and 5 min after NIV ended; it was repeated 60 min later. In group H the acoustic compliance increased after NIV from 2.0+/-.6 ml to 2.3+/ .6 ml ( P<.01), suggesting that the tympanic membrane became less stiff; 1 h later the compliance returned to basal values (2.0+/-.7 ml); in group M the compliance was unaffected (from 2.0+/-.5 ml to 2.0+/-.4 ml; 1.9+/-.4 ml 1 h later). The acoustic reflex, i.e., the contraction of the stapedial muscle in response to an auditory stimulus, involving the acoustic and facial nerves, was also evaluated during impedenzometry at 250 Hz, 500 Hz, 1,000 Hz, and 4,000 Hz; no significant change of the threshold was observed. CONCLUSIONS: The tympanic membrane is tighten by the tensor tympani and a reversible loosening suggests muscle fatigue in response to the application of intermittent positive pressure applied to the external ear during NIV with helmet. The loss of tensor tympani protective action could theoretically predispose the middle and inner ear to mechanical damage during NIV with helmet, suggesting the use of protective devices (ear plugs) in selective cases requiring long-term, high-pressure treatment. PMID- 12589537 TI - Development of CAPS markers based on three key genes of the phenylpropanoid pathway in tea, Camellia sinensis (L.) O. Kuntze, and differentiation between assamica and sinensis varieties. AB - The genetic diversity of tea, Camellia sinensis (L.) O. Kuntze, including the two main cultivated sinensis and assamica varieties, was investigated based on PCR RFLP analysis of PAL, CHS2 and DFR, three key genes involved in catechin and tannin synthesis and directly responsible for tea taste and quality. Polymorphisms were of two types: amplicon length polymorphism (ALP) due to the presence of indels in two introns of PAL and DFR, and point mutations detected after restriction of amplified fragments with appropriate enzymes. A progeny test showed that all markers segregated in a Mendelian fashion and that polymorphisms were exclusively co-dominant. CHS2, which belongs to a multi-gene family, allowed for greater variation than the single-copy PAL gene. Based on Nei's gene diversity index, var. sinensis was revealed to be more variable than var. assamica, and that a higher proportion of overall diversity resided within varieties as compared to between varieties. Even though no specific DNA profile was found for either tea varieties following any single PCR-RFLP analysis, a factorial correspondence analysis carried out on all genotypes and markers separated the tea samples into two distinct groups according to their varietal status. This reflects the large difference between var. sinensis and var. assamica in their polyphenolic profiles. The STS-based markers developed in this study will be very useful in future mapping, population genetics and fingerprinting studies of this important crop species and other Camellia species, as the primers have also proven successful in the three other subgenera of this genus. PMID- 12589538 TI - QTL analysis of genotype x environment interactions affecting cotton fiber quality. AB - Cotton is unusual among major crops in that large acreages are grown under both irrigated and rainfed conditions, making genotype x environment interactions of even greater importance than usual in designing crop-improvement strategies. We describe the impact of well-watered versus water-limited growth conditions on the genetic control of fiber quality, a complex suite of traits that collectively determine the utility of cotton. Fiber length, length uniformity, elongation, strength, fineness, and color (yellowness) were influenced by 6, 7, 9, 21, 25 and 11 QTLs (respectively) that could be detected in one or more treatments. The genetic control of cotton fiber quality was markedly affected both by general differences between growing seasons ("years") and by specific differences in water management regimes. Seventeen QTLs were detected only in the water-limited treatment while only two were specific to the well-watered treatment, suggesting that improvement of fiber quality under water stress may be even more complicated than improvement of this already complex trait under well-watered conditions. In crops such as cotton with widespread use of both irrigated and rainfed production systems, the need to manipulate larger numbers of genes to confer adequate quality under both sets of conditions will reduce the expected rate of genetic gain. These difficulties may be partly ameliorated by efficiencies gained through identification and use of diagnostic DNA markers, including those identified herein. PMID- 12589539 TI - Ecological-genomic diversity of microsatellites in wild barley, Hordeum spontaneum, populations in Jordan. AB - We analyzed the ecological-genomic diversity of microsatellites of wild barley, Hordeum spontaneum (C. Koch) Thell., at 18 loci in 306 individuals of 16 populations from Jordan across a southward transect of increasing aridity. The 18 microsatellites revealed a total of 249 alleles, with an average of 13.8 alleles per locus (range 3-29), with nonrandom distribution. The proportion of polymorphic loci per population averaged 0.91 (range 0.83-1.00); gene diversity, He, averaged 0.512 (range 0.38-0.651). We compared the number of alleles of the 18 loci to those found in Israel populations by Turpeinen et al. Out of the 280 alleles, 138 (49.3%) were unique (i.e. occurred in only one of the countries). The percentage of unique alleles in Jordan and Israel populations was 43.0% and 17.9%, respectively, suggesting that Jordan is an important center of origin and diversity of wild barley. Estimates of mean gene diversity were highest in the populations collected near the Golan Heights, such as Shuni North, Shuni South and Jarash. Sixty nine percent of the microsatellite variation was partitioned within populations and 31% between populations. Associations between ecogeographical values and gene diversity were established for eight microsatellite loci. The cluster produced by simple sequence repeat (SSR) data is mostly coincidence with the result of the dendrogram of the Spalax ehrenbergi superspecies of subterranean mole rats in Jordan based on allozyme gene loci. The major soil type in the wild barley habitat of each ecological group was different. Stepwise multiple regression analysis indicated that the variance of gene diversity was explained by altitude (R(2) = 0.362**). These observations suggest that microsatellites are at least partly adaptive and subject to natural selection. PMID- 12589540 TI - Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.). AB - A software tool was developed for the identification of simple sequence repeats (SSRs) in a barley ( Hordeum vulgare L.) EST (expressed sequence tag) database comprising 24,595 sequences. In total, 1,856 SSR-containing sequences were identified. Trimeric SSR repeat motifs appeared to be the most abundant type. A subset of 311 primer pairs flanking SSR loci have been used for screening polymorphisms among six barley cultivars, being parents of three mapping populations. As a result, 76 EST-derived SSR-markers were integrated into a barley genetic consensus map. A correlation between polymorphism and the number of repeats was observed for SSRs built of dimeric up to tetrameric units. 3'-ESTs yielded a higher portion of polymorphic SSRs (64%) than 5'-ESTs did. The estimated PIC (polymorphic information content) value was 0.45 +/- 0.03. Approximately 80% of the SSR-markers amplified DNA fragments in Hordeum bulbosum, followed by rye, wheat (both about 60%) and rice (40%). A subset of 38 EST derived SSR-markers comprising 114 alleles were used to investigate genetic diversity among 54 barley cultivars. In accordance with a previous, RFLP-based, study, spring and winter cultivars, as well as two- and six-rowed barleys, formed separate clades upon PCoA analysis. The results show that: (1) with the software tool developed, EST databases can be efficiently exploited for the development of cDNA-SSRs, (2) EST-derived SSRs are significantly less polymorphic than those derived from genomic regions, (3) a considerable portion of the developed SSRs can be transferred to related species, and (4) compared to RFLP-markers, cDNA SSRs yield similar patterns of genetic diversity. PMID- 12589541 TI - Mapping QTL for popping expansion volume in popcorn with simple sequence repeat markers. AB - Popping expansion volume is the most important quality trait in popcorn ( Zea mays L.), but its genetics is not well understood. The objectives of this study were to map quantitative trait loci (QTLs) responsible for popping expansion volume in a popcorn x dent corn cross, and to compare the predicted efficiencies of phenotypic selection, marker-based selection, and marker-assisted selection for popping expansion volume. Of 259 simple sequence repeat (SSR) primer pairs screened, 83 pairs were polymorphic between the H123 (dent corn) and AG19 (popcorn) parental inbreds. Popping test data were obtained for 160 S(1) families developed from the [AG19(H123 x AG19)] BC(1) population. The heritability ( h(2)) for popping expansion volume on an S(1) family mean basis was 0.73. The presence of the gametophyte factor Ga1(s) in popcorn complicates the analysis of popcorn x dent corn crosses. But, from a practical perspective, the linkage between a favorable QTL allele and Ga1(s) in popcorn will lead to selection for the favorable QTL allele. Four QTLs, on chromosomes 1S, 3S, 5S and 5L, jointly explained 45% of the phenotypic variation. Marker-based selection for popping expansion volume would require less time and work than phenotypic selection. But due to the high h(2) of popping expansion volume, marker-based selection was predicted to be only 92% as efficient as phenotypic selection. Marker-assisted selection, which comprises index selection on phenotypic and marker scores, was predicted to be 106% as efficient as phenotypic selection. Overall, our results suggest that phenotypic selection will remain the preferred method for selection in popcorn x dent corn crosses. PMID- 12589542 TI - Genetics of resistance to anthracnose and identification of AFLP and RAPD markers linked to the resistance gene in PI 320937 germplasm of lentil (Lens culinaris Medikus). AB - Anthracnose, caused by Colletotrichum truncatum, is a major disease problem and production constraint of lentil in North America. The research was conducted to examine the resistance to anthracnose in PI 320937 lentil and to identify molecular markers linked to the resistance gene in a recombinant inbred line (RIL) population developed from a cross of Eston lentil, the susceptible parent, and PI 320937, the resistant parent. A total of 147 F(5:6) RILs were evaluated for resistance to anthracnose in the greenhouse using isolate 95B36 of C. truncatum. Bulked segregant analysis (BSA) strategy was employed and two contrasting DNA bulks were constructed based on greenhouse inoculation of F(5) derived F(6) RILs. DNA from the parents and bulks were screened with 700 RAPD primers and seven AFLP primer combinations. Analysis of segregation data indicated that a major dominant gene was responsible for resistance to anthracnose while variations in the resistance level among RILs could be the influences of minor genes. We designate the major gene as LCt-2. MapMaker analysis produced two flanking RAPD markers OPEO6(1250) and UBC-704(700) linked to LCt-2 locus in repulsion (6.4 cM) and in coupling (10.5 cM), respectively. Also, three AFLP markers, EMCTTACA(350) and EMCTTAGG(375) in coupling, and EMCTAAAG(175) in repulsion, were linked to the LCt-2 locus. These markers could be used to tag the LCt-2 locus and facilitate marker-assisted selection for resistance to anthracnose in segregating populations of lentil in which PI 320937 was used as the source of resistance. Also, a broader application of the linked RAPD markers was also demonstrated in Indianhead lentil, widely used as a source of resistance to anthracnose in the breeding program at the Crop Development Centre, University of Saskatchewan. Further selection within the few F(5:6) lines should be effective in pyramiding one or several of the minor genes into the working germplasm of lentil, resulting in a more durable and higher level of resistance. PMID- 12589543 TI - Simple sequence repeat (SSR) analysis for assessment of genetic variability in apricot germplasm. AB - Thirty SSR primer combinations, developed from peach SSR-enriched genomic libraries and BAC libraries of peach [ Prunus persica (L.) Batsch.], were tested for cross amplification with 74 apricot ( Prunus armeniaca L.) germplasm accessions. Twelve primer pairs amplified 14 polymorphic SSR loci useful for discriminating most apricot cultivars, as well as for investigating patterns of variation in apricot germplasm. Levels of polymorphism were higher than the levels described using other codominant marker systems (i.e., isozymes, RFLP markers). Overall, 107 alleles were identified, and all but 11 accessions were unambiguously discriminated. Genetic differentiation of native germplasm into traditional ecogeographical groups was low, with a high level of genetic identity (> 0.75) between the groups. However, neighbor joining cluster analysis of marker distances between cultivars reflected the complex history of apricot domestication, producing groupings not evidently based on the geographical origin of the cultivars. Distant positioning of Chinese cultivars on UPGMA and neighbor joining dendrograms supports the authors' consideration of Chinese apricots as subspecies, Prunus armeniaca var. ansu Maxim., rather than a separate species. PMID- 12589544 TI - Soybean germplasm pools in Asia revealed by nuclear SSRs. AB - Soybean was domesticated in East Asia, where various kinds of landraces have been established as a result of adaptation to different environments and the diversification of food cultures. Asia is thus an important germplasm pool of soybean. In order to evaluate the genetic structure of the Asian soybean population, we analyzed allelic profiles at 20 simple-sequence repeat (SSR) loci of 131 accessions introduced from 14 Asian countries. The SSR loci produced an average of 11.9 alleles and a mean gene diversity of 0.782 in the accessions tested. Quantification theory III analysis and cluster analysis with the UPGMA method clearly separated the Japanese from the Chinese accessions, suggesting that the Japanese and Chinese populations formed different germplasm pools. The Korean accessions were involved in both germplasm pools, whereas most of the accessions from southeast and south/central Asia were derived from the Chinese pool. Relatively high genetic diversity and the absence of region-specific clusters in the southeast and south/central Asian populations suggest that soybean in these areas has been introduced repeatedly and independently from the diverse Chinese germplasm pool. The present study indicates that the two germplasm pools can be used as exotic genetic resources to enlarge the genetic bases of the respective Asian soybean populations. PMID- 12589545 TI - The tomato dark green mutation is a novel allele of the tomato homolog of the DEETIOLATED1 gene. AB - A comprehensive, multi-generation, allele test, carried out in this study, suggests that the tomato mutations dark-green (dg) and high pigment 2(j) (hp 2(j)) are allelic. The hp-2(j) mutant is caused by a mutation in the tomato homolog of the DEETIOLATED1 (DET1) gene, involved in the signal transduction cascade of light perception and morphogenesis. This suggestion is in agreement with the exaggerated photomorphogenic de-etiolation response of homozygous dg mutants grown under modulated light conditions. Sequence analysis of the DET1 gene was carried out in dg mutants representing two different lines, and revealed a single A-to-T base transversion in the second exon of the DET1 gene in comparison with the normal wild-type sequence. This transversion results in a conserved Asparagine(34)-to-Isoleucine(34) amino-acid substitution, and eliminates a recognition site for the AclI restriction endonuclease, present in the wild-type and in the other currently known tomato mutants at the DET1 locus. This polymorphism was used to develop a PCR-based DNA marker, which enables an early genotypic selection for breeding lycopene-rich tomatoes. Using this marker and sequence analysis we demonstrate that an identical base transversion also exists in dg mutants of the cultivar Manapal, in which the natural dg mutation was originally discovered. A linkage analysis, carried out in a F(2) population, shows a very strong linkage association between the DET1 locus of dg mutant plants and the photomorphogenic response of the seedlings, measured as hypocotyl length (12 < LOD Score < 13, R(2) = 51.1%). The results presented in this study strongly support the hypothesis that the tomato dg mutation is a novel allele of the tomato homolog of the DET1 gene. PMID- 12589546 TI - Inheritance and fine mapping of fertility restoration for cytoplasmic male sterility in Gossypium hirsutum L. AB - Genetics of CMS fertility restoration was presented through the analysis of classic genetics and molecular markers. Based on F(2) segregation of the crosses between CMS and the restoring lines, the testcrosses and F(1) x F(1) populations, together with RAPD and SSR mapping, one dominant gene was identified to control the CMS fertility restoration in cotton. The strategy of genotype representation analysis (GRA) was put forward to screen the markers linked with the Rf(1) locus. Using 1,025 random decamer primers and 282 pairs of SSR primers, two RAPD and three SSR markers were identified to be closely linked to the Rf(1) gene. Among the five markers, three were co-dominantly inherited. Additionally, based on the analysis of monosomic and telesomic lines with one SSR maker, the Rf(1) locus could be located on the long arm of chromosome 4. The molecular markers available here are helpful in the development of the elite restoring lines in cotton by marker-assisted selection. PMID- 12589547 TI - Microsatellite DNA and RAPD fingerprinting, identification and genetic relationships of hybrid poplar (Populus x canadensis) cultivars. AB - Microsatellite DNA markers of ten SSR loci and 248 RAPD loci (resolved by 26 RAPD primers) were used for DNA fingerprinting and differentiation of 17 widely grown Populus x canadensis syn. Populus x euramericana (interspecific Populus deltoides x Populus nigra hybrids) cultivars ("Baden 431", "Blanc du Poitou", "Canada Blanc", "Dorskamp 925", "Eugenei", "Gelrica", "Grandis", "Heidemij", "I-55/56", "I-132/56", "I-214", "Jacometti", "Ostia", "Regenerata", "Robusta", "Steckby" and "Zurich 03/3"), and determination of their genetic interrelationships. Informativeness of microsatellite and RAPD markers was also evaluated in comparison with allozyme markers for clone/cultivar identification in P. x canadensis. High microsatellite DNA and RAPD genetic diversity was observed in the sampled cultivars. All of the 17 P. x canadensis cultivars could be differentiated by their multilocus genotypes at four SSR loci, and were heterozygous for their parental species-specific alleles at the PTR6 SSR locus. Except for "Canada Blanc" and "Ostia", which had identical RAPD patterns, all cultivars could also be differentiated by RAPD fingerprints produced by each of the two RAPD primers, OPA07 and OPB15. For microsatellites, the mean number of alleles, polymorphic information content, observed heterozygosity, observed number of genotypes and the number of cultivars with unique genotypes per locus was 5.2, 0.64, 0.67, 5.7 and 2.2, respectively. For RAPD markers, the number of haplotypes per locus, and the number of cultivars with unique RAPD profiles per locus were 1.06 and 0.72, respectively. Overall, microsatellite DNA markers were the most informative for DNA fingerprinting of P. x canadensis cultivars. On the per locus basis, microsatellites were about six-times more informative than RAPD markers and about nine-times more informative than allozyme markers. However, on the per primer basis, RAPD markers were more informative. The UPGMA cluster plots separated the 17 cultivars into two major groups based on their microsatellite genotypic similarities, and into three major groups based on their RAPD fragment similarities. Both the microsatellite and RAPD data suggest that the cultivars "Baden 431", "Heidemij", "Robusta" and "Steckby" are genetically closely related. The inter-cultivar genetic relationships from microsatellite DNA and RAPD markers were consistent with those observed from allozyme markers, and were in general agreement with their speculated origin. Microsatellite DNA and RAPD markers could be used for clone and cultivar identification, varietal control and registration, and stock handling in P. x canadensis. PMID- 12589548 TI - The heat-stable root-knot nematode resistance gene Mi-9 from Lycopersicon peruvianum is localized on the short arm of chromosome 6. AB - The tomato gene Mi-1 confers resistance to three species of root-knot nematodes, Meloidogyne spp. However, the resistance mediated by Mi-1 is inactive at soil temperatures above 28 degrees C. Previously, we identified and mapped a novel heat-stable nematode resistance gene from the wild species Lycopersicon peruvianum accession LA2157 on to chromosome 6. Here we report further characterization of this heat-stable resistance against three Mi-1-avirulent biotypes of Meloidogyne javanica, Meloidogyne arenaria and Meloidogyne incognita. Screening segregating F(2) and F(3) progenies, derived from an intraspecific cross between susceptible LA392 and resistant LA2157, for nematode resistance at 25 degrees C and 32 degrees C, revealed a simple dominant monogenic inheritance with all the biotypes tested. We designate this gene as Mi-9. As a first step towards cloning of Mi-9, we constructed a linkage map around this gene. A total of 216 F(2) progeny from the cross between LA392 and LA2157 were screened with M. javanica at 32 degrees C and with CT119 and Aps-1, markers that flank the genetic interval that contains the Mi-1 gene. DNA marker analysis indicated that these markers also flank Mi-9. Further mapping of recombinants with both RFLP and PCR based markers localized Mi-9 to the short arm of chromosome 6 and within the same genetic interval that spans the Mi-1 region. PMID- 12589549 TI - Saturation of two chromosome regions conferring resistance to SCMV with SSR and AFLP markers by targeted BSA. AB - Quantitative trait loci (QTLs) and bulked segregant analyses (BSA) identified the major genes Scmv1 on chromosome 6 and Scmv2 on chromosome 3, conferring resistance against sugarcane mosaic virus (SCMV) in maize. Both chromosome regions were further enriched for SSR and AFLP markers by targeted bulked segregant analysis (tBSA) in order to identify and map only markers closely linked to either Scmv1 or Scmv2. For identification of markers closely linked to the target genes, symptomless individuals of advanced backcross generations BC5 to BC9 were employed. All AFLP markers, identified by tBSA using 400 EcoRI/ MseI primer combinations, mapped within both targeted marker intervals. Fourteen SSR and six AFLP markers mapped to the Scmv1 region. Eleven SSR and 18 AFLP markers were located in the Scmv2 region. Whereas the linear order of SSR markers and the window size for the Scmv2 region fitted well with publicly available genetic maps, map distances and window size differed substantially for the Scmv1 region on chromosome 6. A possible explanation for the observed discrepancies is the presence of two closely linked resistance genes in the Scmv1 region. PMID- 12589550 TI - Deficiency of a cytosolic ascorbate peroxidase associated with chilling tolerance in soybean. AB - We investigated the isozyme profiles of antioxidant enzymes in cultivars and lines with different seed productivity in cool climate conditions as a step towards understanding the physiological and genetical mechanisms underlying chilling tolerance in soybean. While no difference in superoxide dismutase, or catalase isozyme profiles was observed among the cultivars and lines tested, we found polymorphism in the ascorbate peroxidase isozyme profile; there were two types, with or without a cytosolic isoform (APX1). The cultivars and lines lacking APX1 proved more tolerant to chilling temperatures, as evaluated by yielding ability. The genotype-dependent deficiency of APX1 was consistent in plants and tissues under various oxidative stress conditions including the exposure to low-temperatures. In addition, the genetic analysis of progeny derived from crossing between cultivars differing in the isozyme profile indicated that the APX1 deficiency is controlled by a single recessive gene (apx1), and is inherited independently of the genes that have previously been identified for their association with chilling tolerance. Molecular and linkage analyses suggested that the variant gene of the APX1-absent genotype coding for a cytosolic APX, which contained a single nucleotide substitution and a single nucleotide deletion in the coding region, is responsible for the genotype dependent deficiency of APX1. The association of APX1 deficiency with chilling tolerance is discussed in detail. PMID- 12589552 TI - Mapping and characterisation of QTL x E interactions for traits determining grain and stover yield in pearl millet. AB - A mapping population of 104 F(3) lines of pearl millet, derived from a cross between two inbred lines H 77/833-2 x PRLT 2/89-33, was evaluated, as testcrosses on a common tester, for traits determining grain and stover yield in seven different field trials, distributed over 3 years and two seasons. The total genetic variation was partitioned into effects due to season (S), genotype (G), genotype x season interaction (G x S), and genotype x environment-within-season interaction [G x E(S)]. QTLs were determined for traits for their G, G x S, and G x E(S) effects, to assess the magnitude and the nature (cross over/non-crossover) of environmental interaction effects on individual QTLs. QTLs for some traits were associated with G effects only, while others were associated with the effects of both G and G x S and/or G, G x S and G x E(S) effects. The major G x S QTLs detected were for flowering time (on LG 4 and LG 6), and mapped to the same intervals as G x S QTLs for several other traits (including stover yield, harvest index, biomass yield and panicle number m(-2)). All three QTLs detected for grain yield were unaffected by G x S interaction however. All three QTLs for stover yield (mapping on LG 2, LG 4 and LG 6) and one of the three QTLs for grain yield (mapping on LG 4) were also free of QTL x E(S) interactions. The grain yield QTLs that were affected by QTL x E(S) interactions (mapping on LG 2 and LG 6), appeared to be linked to parallel QTL x E(S) interactions of the QTLs for panicle number m(-2) on (LG 2) and of QTLs for both panicle number m(-2) and harvest index (LG 6). In general, QTL x E(S) interactions were more frequently observed for component traits of grain and stover yield, than for grain or stover yield per se. PMID- 12589551 TI - Characterization of a unique genomic clone located 5' upstream of the Oshsp16.9B gene on chromosome 1 in rice (Oryza sativa L. cv Tainung No. 67). AB - Small heat-shock proteins (sHSP) are the most abundant heat stress-induced proteins in plants. In rice, there are at least seven members of class-I sHSP. A 1.6-kb DNA fragment was isolated from the EcoRI-digested rice genomic library probed with the cDNA pTS1 encoding a 16.9-kDa class-I sHSP. This fragment was composed of 365-bp tandem direct repeats (DRs) and 441-bp near perfect long terminal inverted repeats (LTIRs). The DRs contain 123-bp regions with 99% nucleotide identity to the 5' coding region of the Oshsp16.9B gene. Two putative pseudogenes were deduced from the DRs. Using the LTIR as a specific probe, Southern-blotting analysis showed that there was a single copy of this 1.6-kb DNA fragment in the rice genome. By genomic walking, we located this fragment in proximity 5'-upstream of the Oshsp16.9B gene that was mapped on chromosome 1 with other two class-I sHSP genes, Oshsp16.9A and Oshsp16.9C. By comparative analysis of the nucleotide sequences of class-I sHSP genes clustered on chromosome 1 between Tainung No. 67 and Nipponbare cultivars, we confirmed our mapping results of these genes and only the promoter region of Oshsp16.9B was different. However, we found that the expression profile of Oshsp16.9B upon different heat stresses in Nipponbare was not significantly different relative to that in Tainung No. 67. PMID- 12589553 TI - The use of molecular markers for germplasm management in a French olive collection. AB - With more than 100 accessions, the CBNMP olive collection includes a major part of the French germplasm. We used molecular markers to characterise all accessions and to study genetic relationships between cultivars. Firstly, 497 olive trees were genotyped using 32 RAPD markers. We identified 114 RAPD profiles and detected several cases of mislabelling, synonymy and homonymy. Secondly, for each RAPD profile, one tree was analysed using mtDNA RFLPs to determine the cytoplasmic lineage of each cultivar and using five nuclear SSR loci. French germplasm displayed ME1, MOM and MCK mitotypes with ME1 prevailing (84%). Based on SSR markers, we revealed a slight differentiation between French cultivars growing in the West and the East side of the Rhone Valley. This study allowed us to construct a molecular data-base for the reference collection and to analyse genetic diversity for further prospecting, and for introducing new olive accessions. PMID- 12589554 TI - An update of the Courtot x Chinese Spring intervarietal molecular marker linkage map for the QTL detection of agronomic traits in wheat. AB - We made an update of the intervarietal molecular marker linkage map of the wheat genome developed using a doubled-haploid (DH) population derived from the cross between the cultivars "Courtot" and "Chinese Spring". This map was constructed using 187 DH lines and 659 markers. The genome was well covered (more than 95%) except for chromosomes from homoeologous group 4 and chromosomes 5D and 7D, which had gaps slightly larger than 50 cM. A core-map based on a set of 200 anchor loci (one marker each 18.4 cM) was developed. The total length of this map was 3,685 cM which is similar to the size of the international reference map of the ITMI population (3,551 cM). Map coverage was identical for the three genomes (A, B and D) and for the number of anchor loci, as well as for the size of the map. Using this map, QTLs for several agronomic traits were detected on phenotypic data from the population grown in Clermont-Ferrand (France) under natural field conditions over 6 years, and in Norwich (UK) in controlled conditions and under natural field conditions in 1 year. Almost all of the 21 chromosomes were involved in at least one trait. However, several regions seemed to contain gene clusters either for grain traits (and thus bread-making quality) or plant development traits. PMID- 12589555 TI - Inheritance of microsatellite alleles in pedigrees of Latvian barley varieties and related European ancestors. AB - Genetic diversity and inheritance of 65 microsatellite (SSR) loci were studied in a set of 37 barley varieties involved in the pedigrees of seven Latvian barley varieties: Abava, Agra, Balga, Imula, Linga, Priekulu 1 and Stendes. Cluster analysis divided all the varieties into two large groups according to their geographic distribution. Moravian, Swedish and Danish varieties clustered separately from varieties from Norway and Finland. The pattern of subgroups of both European and Latvian varieties was in accordance with their pedigree information. Graphical genotypes of microsatellite alleles of all seven barley chromosomes were determined for all the 37 varieties studied. Parental inheritance and transmission of microsatellite alleles through the generations of the pedigrees were analysed. The results confirmed the importance and informative value of microsatellite markers for genetic studies in barley and their utility for barley breeding and other applications in fundamental and applied barley genetics. PMID- 12589556 TI - Temporal flux in the morphological and molecular diversity of UK barley. AB - Genetic-diversity assessments, using both phenotypic and molecular-marker data, were made on a collection of 134 barley varieties (both winter and spring types), chosen on the basis of their representation on the NIAB "Recommended List" over the period 1925-1995. Genotypic (AFLP and SSR) and phenotypic (UPOV characters) data were analysed to determine short- and long-term temporal trends in diversity over the period. A consistent pattern emerged demonstrating that only a minor proportion of the overall variance appears to be the result of any temporal drift, although there were strong indications of qualitative shifts in diversity, probably related to the changing relative acreage of winter and spring barleys over the study period. Our overall conclusions are that systematic plant breeding does not inevitably lead to a reduction in the genetic diversity of agricultural crops, and that diverse breeding programmes and the variety delivery systems in place in the UK have generally been successful in maintaining sufficient genetic diversity to allow the steady rise in genetic potential that has been a feature of 20th century crop breeding. The concentration of breeding effort into a smaller number of independent programmes is likely to be prejudicial to the maintenance of the genetic diversity of a crop. PMID- 12589557 TI - Prediction of hybrid performance in grain sorghum using RFLP markers. AB - Heterosis is an important component of hybrid yield performance. Identifying high yielding hybrids is expensive and involves testing large numbers of hybrid combinations in multi-environment trials. Molecular marker diversity has been proposed as a more efficient method of selecting superior combinations. The aim of this study was to investigate the value of molecular marker-based distance information to identify high yielding grain sorghum hybrids in Australia. Data from 48 trials were used to produce hybrid performance-estimates for four traits (yield, height, maturity and stay green) for 162 hybrid combinations derived from 70 inbred parent lines. Each line was screened with 113 mapped RFLP markers. The Rogers distances between the parents of each hybrid were calculated from the marker information on a genome basis and individually for each of the ten linkage groups of sorghum. Some of the inbred parents were related so the hybrids were classified into 75 groups with each group containing individual hybrids that showed similar patterns of Rogers distances across linkage groups. Correlations between hybrid-group performance and hybrid-group Rogers distances were calculated. A significant correlation was observed between whole genome-based Rogers distance and yield ( r = 0.42). This association is too weak to be of value for identifying superior hybrid combinations. One reason for the generally poor association between parental genetic diversity and yield may be that important QTLs influencing heterosis are located in particular chromosome regions and not distributed evenly over the genome. Variation in the sign and magnitude of correlations between Rogers distance and hybrid-group performance for particular linkage groups observed in this study support this hypothesis. The concept of using diversity on individual linkage groups to predict performance was explored. Using data from just two linkage groups 38% of the variation in hybrid performance for grain yield could be explained. A model combining phenotypic trait data and parental diversity on particular linkage groups explained 71% of the variation in grain yield and has potential for use in the selection of heterotic hybrids. PMID- 12589558 TI - Evaluation of BC2 progenies derived from 3x-2x and 3x-4x crosses of Lilium hybrids: a GISH analysis. AB - An allotriploid (ALA, 2n=3 x=36) BC(1) plant was obtained by backcrossing a diploid F(1) interspecific hybrid (LA, 2n=2 x=24), derived from a Lilium longiflorum (L genome) and an Asiatic hybrid (A genome), to the latter parent. This allotriploid was backcrossed to a diploid Asiatic hybrid (2n=2 x=24) and to an allotetraploid (LLAA, 2n=4 x=48) LA hybrid. A total of 25 plants of these crosses were examined for ploidy level, and 12 individuals were analyzed for their genome constitution through genomic in situ hybridization (GISH). In most cases the progenies from the triploid-diploid (3 x-2 x) crosses consisted of aneuploids. Further more, there was evidence for the formation of near-haploid (x=12+2) to triploid (3 x=36) gametes in the allotriploid BC(1) plant. The progenies of triploid-tetraploid (3 x-4 x) cross also consisted of mostly aneuploids but in this case the triploid female parent had contributed predominantly near-triploid (2n) gametes for the origin of BC(2) progenies. The different ploidy levels observed between 3 x-2 x and 3 x-4 x crosses are possibly caused by preferential fertilization or survival resulting in a different ratio of chromosome numbers between the embryo and endosperm. Though Lilium has a tetrasporic, eight-nucleate type of embryo sac formation (Fritillaria type), the observed difference between the progeny types in 3 x-2 x and 3 x-4 x crosses is comparable to that of observed in monosporic eight nucleate types (Polygonum type) that predominate in most genera of Angiosperms. An important feature of the genome constitution of the progenies was that the homoeologous recombinant chromosomes were transmitted intact from BC(1) to BC(2) progenies in variable numbers. In addition, there was evidence for the occurrence of new homoeologous recombinations in the triploid BC(1). Of the two euploid BC(2) plants one had originated through the parthenogenetic development of a 2n egg and the other had originated through indeterminate meiotic restitution (IMR). PMID- 12589559 TI - Examples of high-frequency EPR studies in bioinorganic chemistry. AB - Low-temperature EPR spectroscopy with frequencies between 95 and 345 GHz and magnetic fields up to 12 T has been used to study metal sites in proteins or inorganic complexes and free radicals. The high-field EPR method was used to resolve g-value anisotropy by separating it from overlapping hyperfine couplings. The presence of hydrogen bonding interactions to the tyrosyl radical oxygens in ribonucleotide reductases were detected. At 285 GHz the g-value anisotropy from the rhombic type 2 Cu(II) signal in the enzyme laccase has its g-value anisotropy clearly resolved from slightly different overlapping axial species. Simple metal site systems with S>1/2 undergo a zero-field splitting, which can be described by the spin Hamiltonian. From high-frequency EPR, the D values that are small compared to the frequency (high-field limit) can be determined directly by measuring the distance of the outermost signal to the center of the spectrum, which corresponds to (2 S-1)* mid R: Dmid R: For example, D values of 0.8 and 0.3 cm(-1) are observed for S=5/2 Fe(III)-EDTA and transferrin, respectively. When D values are larger compared to the frequency and in the case of half-integer spin systems, they can be obtained from the frequency dependence of the shifts of g(eff), as observed for myoglobin in the presence ( D=5 cm(-1)) or absence ( D=9.5 cm(-1)) of fluoride. The 285 and 345 GHz spectra of the Fe(II)-NO-EDTA complex show that it is best described as a S=3/2 system with D=11.5 cm(-1), E=0.1 cm(-1), and g(x)= g(y)= g(z)=2.0. Finally, the effects of HF-EPR on X-band EPR silent states and weak magnetic interactions are demonstrated. PMID- 12589560 TI - Different pathways of radical translocation in yeast cytochrome c peroxidase and its W191F mutant on reaction with H(2)O(2) suggest an antioxidant role. AB - In the absence of exogenous donors, turnover of 10 molar equivalents of H(2)O(2) by wild-type recombinant cytochrome c peroxidase [CCP(MI)] and its W191F mutant at pH 7.0 occurs by oxidation of endogenous donors on the polypeptide. No O(2) evolution was observed with either enzyme on reaction with 10 molar equivalents of H(2)O(2), eliminating catalase-like activity, but O(2) evolution was observed when 100 molar equivalents of H(2)O(2) were added to the enzymes. Protein dimers were observed by SDS-PAGE following H(2)O(2) turnover by the peroxidases, and dimeric forms of CCP(MI) and CCP(W191) were isolated by gel-permeation chromatography. LC-ESI-MS analysis of the tryptic digests of the dimers revealed the previously reported T(6)-T(6) crosslink and a new crosslink between T(6) T(26), but no T(26)-T(26) crosslink. The crosslinked tryptic peptides contain the exposed tyrosine residues Tyr36, Tyr39 and Tyr42 (T(6)), and Tyr229 and Tyr236 (T(26)). Addition of a spin trap, 2-methyl-2-nitrosopropane (MNP), to the CCP(MI)/H(2)O(2) reaction resulted in MNP labeling of peptides T(6), T(21) (which contains Tyr153) and T(26). MNP labeling of Tyr236 was found by sequencing peptide T(26). MNP labeling did not compete with dimerization of H(2)O(2) oxidized CCP(W191F), suggesting that dityrosine formation in this mutant is very rapid owing to the high reactivity of radicals formed on T(6). H(2)O(2)-dependent formation of CCP-cytochrome c heterodimers was observed for both CCP(MI) and W191F in the presence of ferricytochrome c, the oxidized form of CCP's donor substrate. Interestingly, no H(2)O(2)-dependent cytochrome crosslinking to the W51F mutant was observed, even though this mutant underwent extensive homocrosslinking. The translocation of oxidizing equivalents from the heme to the surface residues of CCP is discussed in terms of an antioxidant role for CCP. PMID- 12589561 TI - A new biomineral identified in the cores of teeth from the chiton Plaxiphora albida. AB - The hydrated iron(III) oxide limonite is reported for the first time as a biomineral. In situ laser Raman spectra of the tooth cores from major lateral teeth of the chiton Plaxiphora albida are compared with those of synthetic and mineral iron phosphates and iron oxides. Raman spectra measured on iron phosphate and iron oxide standard materials are shown to be easily distinguishable from one another. The central tooth cores of mature P. albida teeth do not show any evidence for the presence of a separate iron phosphate mineral. Rather, in each tooth a narrow band of the hydrated iron(III) oxide limonite is shown to separate the magnetite of the tooth surface from a central core region comprising both lepidocrocite and limonite. The high concentration of phosphorus in P. albida tooth cores, previously observed by energy dispersive spectroscopy, is not associated with a separate iron phosphate mineral, indicating that this element may be adsorbed onto the surface of the iron oxide minerals present. The failure to detect a separate iron(III) phosphate is discussed with reference to other chiton species that display high levels of iron and phosphorus in the cores of their mature major lateral teeth. PMID- 12589562 TI - 4-nitrocatechol as a probe of a Mn(II)-dependent extradiol-cleaving catechol dioxygenase (MndD): comparison with relevant Fe(II) and Mn(II) model complexes. AB - Mn(II)-dependent 3,4-dihydroxyphenylacetate 2,3-dioxygenase (MndD) is an extradiol-cleaving catechol dioxygenase from Arthrobacter globiformis that has 82% sequence identity to and cleaves the same substrate (3,4 dihydroxyphenylacetic acid) as Fe(II)-dependent 3,4-dihydroxyphenylacetate 2,3 dioxygenase (HPCD) from Brevibacterium fuscum. We have observed that MndD binds the chromophoric 4-nitrocatechol (4-NCH(2)) substrate as a dianion and cleaves it extremely slowly, in contrast to the Fe(II)-dependent enzymes which bind 4-NCH(2) mostly as a monoanion and cleave 4-NCH(2) 4-5 orders of magnitude faster. These results suggest that the monoanionic binding state of 4-NC is essential for extradiol cleavage. In order to address the differences in 4-NCH(2) binding to these enzymes, we synthesized and characterized the first mononuclear monoanionic and dianionic Mn(II)-(4-NC) model complexes as well as their Fe(II)-(4-NC) analogs. The structures of [(6-Me(2)-bpmcn)Fe(II)(4-NCH)](+), [(6-Me(3) TPA)Mn(II)(DBCH)](+), and [(6-Me(2)-bpmcn)Mn(II)(4-NCH)](+) reveal that the monoanionic catecholate is bound in an asymmetric fashion (Delta r(metal O(catecholate))=0.25-0.35 A), as found in the crystal structures of the E(.)S complexes of extradiol-cleaving catechol dioxygenases. Acid-base titrations of [(L)M(II)(4-NCH)](+) complexes in aprotic solvents show that the p K(a) of the second catecholate proton of 4-NCH bound to the metal center is half a p K(a) unit higher for the Mn(II) complexes than for the Fe(II) complexes. These results are in line with the Lewis acidities of the two divalent metal ions but are the opposite of the trend observed for 4-NCH(2) binding to the Mn(II)- and Fe(II) catechol dioxygenases. These results suggest that the MndD active site decreases the second p K(a) of the bound 4-NCH(2) relative to the HPCD active site. PMID- 12589563 TI - The importance of porphyrin distortions for the ferrochelatase reaction. AB - Ferrochelatase is the terminal enzyme in haem biosynthesis, i.e. the enzyme that inserts a ferrous ion into the porphyrin ring. Suggested reaction mechanisms for this enzyme involve a distortion of the porphyrin ring when it is bound to the enzyme. We have examined the energetics of such distortions using various theoretical calculations. With the density functional B3LYP method we calculate how much energy it costs to tilt one of the pyrrole rings out of the porphyrin plane for an isolated porphyrin molecule without or with a divalent metal ion in the centre of the ring. A tilt of 30 degrees costs 65-130 kJ/mol for most metal ions, but only approximately 48 kJ/mol for free-base (neutral) porphine. This indicates that once the metal is inserted, the porphyrin becomes stiffer and flatter, and therefore binds with lower affinity to a site designed to bind a distorted porphyrin. This would facilitate the release of the product from ferrochelatase. This proposal is strengthened by the fact that the only tested metal ion with a lower distortion energy than free-base porphyrin (Cd(2+)) is an inhibitor of ferrochelatase. Moreover, it costs even less energy to tilt a doubly deprotonated porphine(2-) molecule. This suggests that the protein may lower the acid constant of the pyrrole nitrogen atoms by deforming the porphyrin molecule. We have also estimated the structure of the protoporphyrin IX substrate bound to ferrochelatase using combined quantum chemical and molecular mechanics calculations. The result shows that the protein may distort the porphyrin by approximately 20 kJ/mol, leading to a distinctly non-planar structure. All four pyrrole rings are tilted out of the porphyrin mean plane (1-16 degrees ) but most towards the putative binding site of the metal ion. The predicted tilt is considerably smaller than that observed in the crystal structure of a porphyrin inhibitor. PMID- 12589564 TI - Evaluating hydrogen bond interactions in enzymes containing Mn(III)-histidine complexation using manganese-imidazole complexes. AB - It is often difficult to control hydrogen bond interactions in small molecule compounds that model metalloenzyme active sites. The imidazole-containing ligands 4,5-dicarboxyimidazole (H(3)DCBI) and 4,5-dicarboxy- N-methylimidazole (H(2)MeDCBI) allow examination of the effects of internal hydrogen bonding between carboxylate and imidazole nitrogen atoms. A new series of mononuclear manganese imidazole complexes have been prepared using these ligands: Mn(III)(salpn)(H(2)DCBI)(DMF) (1), Mn(III)(salpn)(HMeDCBI) (2), Mn(III)(dtsalpn)(HMeDCBI) (3), [Mn(IV)(dtsalpn)(HMeDCBI)]PF(6) (4), Mn(III)(salpn)(H(2)DCBI) (5), Mn(III)(dtsalpn)(H(2)DCBI) (6), and Mn(IV)(dtsalpn)(H(2)DCBI)PF(6) (8). Complexes 1, 2, 3, 5, and 6 have been prepared by direct reaction of salpn [salpn=(salicylideneaminato)-1,3 diaminopropane)] or dtsalpn [dtsalpn=(3,5-di- t-butylsalicylideneaminato)-1,3 diaminopropane)] and H(3)DCBI and H(2)MeDCBI with Mn(III) acetate, while complexes 4 and 8 were made by bulk electrolysis of complex 3 or 6 in dichloromethane. Complexes 1, 2, and 6 were characterized by X-ray diffraction. The impact of hydrogen bonding interactions of the complexes has been demonstrated by X-ray diffraction, cyclic voltammetry, and EPR spectroscopy. In all complexes the central metal ion is present in a six-coordinate geometry. Magnetic susceptibility measurements confirm the spin and oxidation states of the complexes. The cyclic voltammograms of 3 and 6 in dichloromethane reveal single, reversible redox waves with E(1/2)=600 mV and 690 mV, respectively. The X-band EPR spectrum of 4 shows a broad signal around g=4.4, and the corresponding complex 8 possesses a broad signal at slightly lower field ( g=5.5) than 4. These studies demonstrate that even small changes in the effective charge of the imidazole ligand can have a profound impact on the structure, spectroscopy, and magnetism of manganese(IV) complexes. We use these observations to present a model that may explain the origin of the g=4.1 signal in the S(2) state of photosystem II. PMID- 12589566 TI - A thin-film electrochemical study of the "blue" copper proteins, auracyanin A and auracyanin B, from the photosynthetic bacterium Chloroflexus aurantiacus: the reduction potential as a function of pH. AB - The reversible formal potentials of auracyanin A and auracyanin B, two closely related "blue" copper proteins from the photosynthetic bacterium Chloroflexus aurantiacus, have been determined by protein film voltammetry in the range 490%). The PPV of the "putaminal" score was high (70%) for the differential diagnosis of MSA with PSP and CBD. The "midbrain" score was significantly higher in PSP and CBD compared to MSA. These results are in accordance with the underlying pathology found in these disorders and demonstrate that a simple MRI scoring procedure may help the neurologist to differentiate primary causes of parkinsonism in everyday practice. PMID- 12589576 TI - Attenuation of methamphetamine induced dopaminergic neurotoxicity by flupirtine: microdialysis study on dopamine release and free radical generation. AB - Flupirtine is a triaminopyridine derived centrally acting analgetic, which has been found to display neuroprotective effects in models of excitotoxic cell damage, global, and focal ischemia, but no direct interaction with any component of the N-methyl-D-aspartate (NMDA) and glutamate triggered Ca(2+)-channel. Additionally flupirtine shows potent antioxidant effects in isolated mitochondria and cell culture. Work in models of monoamine depletion and neuroleptic induced catalepsy in rats suggests a interaction of flupirtine with the dopaminergic neurotransmitter system as well. This prompted us to examine the effect of flupirtine on methamphetamine toxicity in mice and to investigate the influence on dopamine release and free radical formation in the rat striatum by microdialysis that may explain methamphetamine neurotoxicity. Pretreatment of C57 BL mice with flupirtine (4 x 10 mg/kg) significantly attenuated the striatal dopamine loss after methamphetamine application (4 x 5 mg/kg). In rats, a single injection of 10 mg/kg flupirtine reduced the methamphetamine induced striatal dopamine release by almost 50%, as measured by in vivo microdialysis. Flupirtine, however, did not influence the increase of free radical formation after methamphetamine infusion, which was assayed after infusion of salicylic acid by quantification of 2,3- and 2,5-dihydroxybenzoic acid. This suggests that other mechanisms rather than dopamine metabolism and autoxidation, may contribute to methamphetamine neurotoxicity. PMID- 12589577 TI - An antihypokinesic action of alpha2-adrenoceptors upon MPTP-induced behaviour deficits in mice. AB - The effects of co-administration of either the dopamine precursor, L-Dopa, or the directly-acting, mixed dopamine (DA) agonist, apomorphine, with the alpha adrenoceptor agonists, clonidine and guanfacine, upon the motor activity of hypoactive L-Dopa-tolerant MPTP-treated C57 BL/6 mice were measured in four experiments. In each case, MPTP (2 x 40 mg/kg, s.c., separated by a 24-hr interval) was administered eight-to-ten weeks before behavioural testing. It was found that clonidine co-administered with L-Dopa (20 mg/kg) restored motor activity in a dose- and parameter-related manner: locomotion and total activity were restored by the 1 mg/kg dose, rearing behaviour by the 0.3 and 1 mg/kg doses. The restorative effects of clonidine (1 mg/kg), co-administered with L Dopa, were antagonised completely by pretreatment with yohimbine (1 mg/kg), but not by prazosin (1 mg/kg). Guanfacine (1 mg/kg) co-administered with L-Dopa (20 mg/kg) restored locomotor, but not rearing, behaviour in L-Dopa-tolerant MPTP treated mice. The antikinesic action of guanfacine was antagonised completely by yohimbine (1 mg/kg), but not prazosin (1 mg/kg). Clonidine (1 or 3 mg/kg) co administered with apomorphine (0.1, 0.3, 1.0 or 3.0 mg/kg), directly-acting DA agonist, did not restore motor behaviour in the hypokinesic L-Dopa-tolerant MPTP treated mice. Nor did apomorphine, by itself, affect the motor activity of these animals. Neurochemical analysis indicated marked DA, DOPAC and HVA depletions in the striatum, and to a much lesser extent in the frontal cortex, of MPTP-treated mice. The synergistic antiparkinsonian action of clonidine with L-Dopa, but not apomorphine, in hypokinetic MPTP mice for the restoration of responding to a suprathreshold dose of L-Dopa, to which "wearing-off" had been induced previously, is discussed. PMID- 12589578 TI - Pre-movement motor excitability is reduced ipsilateral to low force pinch grips. AB - Motor excitability ipsilateral to pinch grips was investigated during the pre movement period. Subjects performed right-handed phasic pinch grips with 2% or 20% maximum voluntary contraction (MVC) in response to a visual go-signal. Transcranial magnetic stimulation (TMS) was applied over the right motor cortex at various intervals before the go-signal and 100 msec after movement onset. Motor evoked potentials were recorded from the relaxed left first dorsal interosseous muscle. Immediately prior to and during 2% MVC pinch grips, MEP amplitudes were reduced. In contrast, MEPs obtained by transcranial electrical stimulation tended to be increased, indicating that MEP decreases are mediated at a cortical level. Before and during 20% MVC pinch grips MEP amplitudes were enhanced. TMS delayed reaction time if applied close to the go-signal. We conclude that the motor cortex ipsilateral to low force movements is inhibited prior to and during movement. PMID- 12589580 TI - [Non-invasive screening for renovascular hypertension -- which test?]. PMID- 12589581 TI - [Long-term follow-up of right atrial multilinear high-frequency ablation in the treatment of recurrent paroxysmal atrial fibrillation]. AB - BACKGROUND AND OBJECTIVE: To investigate the long-term follow-up after right atrial compartmentalization using radiofrequency catheter ablation to treat recurrent paroxysmal atrial fibrillation. PATIENTS AND METHODS: 33 patients (eight women / 25 men, mean age 56.1+/-9.9 years) with highly symptomatic recurrent paroxysmal atrial fibrillation and mostly unresponsive to antiarrhythmic drugs were enrolled in this prospective study. All patients underwent radiofrequency catheter ablation, including right atrial compartmentalization and ablation of the right atrial isthmus region. The primary goal during follow-up was documentation of arrhythmia-related symptoms using a SF 36 quality-of-life questionnaire. RESULTS: During a mean follow-up of 2.1 years 21 % of patients were free of a relapse under continued antiarrhythmic medication, 79 % suffered at least from one period of atrial fibrillation. According to the underlying heart disease patients classified as "lone atrial fibrillation" (40 % without a relapse) showed improvement particularly compared to patients with coronary heart disease (10 % without a relapse). In the group of patients with a relapse of atrial fibrillation the mean of duration of an arrhythmic episode decreased significantly from 10.6 to 2.3 hours under continued administration of antiarrhythmic drugs (p = 0.01), as did the number of episodes, from 2.2 to 1.9/week. CONCLUSION: Despite of the high rate of clinical relapse, patients can profit due to an improved responsiveness to antiarrhythmic drugs after ablation. Right atrial compartmentalization should not be understood as a causal therapy but as an approach to a symptomatic form of hybrid therapy. PMID- 12589582 TI - [Schooling, living conditions and study motivation of students partaking in the traditional or reformed course of instruction at the Charite (Hospital)]. AB - BACKGROUND: There are two different tracks in undergraduate medical education at the Charite, a traditional (TT) one and a reformed one (RT) which was implemented in 1999. We report a survey reflecting on possible differences between students joining the two tracks. METHODS: During the winter term 2001/02 a questionnaire was sent to all 223 newly enrolled medical students. Three groups were distinguished: those following either the traditional or the reformed course (TT or RT), or those who followed the TT, although they had applied for it, because there were not enough vacancies in the RT. PARTICIPANTS: 150 (63% female; average age: 21.5 years) of 223 first year medical students. RESULTS: Applicants for the RT did not differ significantly from those of the other groups with respect to school-leaving grades, their choice of main courses at school an their motivation to study medicine. But they differed in the demands that they placed on their instruction. The wish for a good practical medical training, advancing their individual potential and the ability to select for themselves any instructional priorities were of greater importance for those choosing the RT than those who preferred the TT. More students of the RT than the TT would again have chosen to study medicine and less often stated that they had considered giving up their medical course. CONCLUSION: Students have different expectations concerning medical curricula. The possibility to choose between different ways of studying may help to meet different expectations. PMID- 12589583 TI - [Competitive sports and dilated cardiomyopathy: the case of a 32-year-old soccer player with ventricular tachycardia]. AB - HISTORY AND ADMISSION FINDINGS: A 32-year-old competitive soccer player presented with palpitations he had felt for 4 weeks during maximal activity (soccer training and match). The physical examination and an exercise electrocardiogram were carried out by his general practitioner up to 19 s at 350 W and a heart rate of 147/min without showing any abnormalities. INVESTIGATIONS: All blood parameters revealed no signs of illness. During treadmill exercise at a heart rate of 181/min, a non-sustained ventricular tachycardia was induced. Echocardiography showed a dilated left ventricle with an enddiastolic diameter of 70 mm and low fractional shortening (28 %). Cardiac catheterization demonstrated a diminished left ventricular ejection fraction (38 %) and an enlarged enddiastolic volume (199 ml) without signs of coronary artery disease. Electrophysiologic testing induced a non-sustained ventricular tachycardia. DIAGNOSIS, TREATMENT AND COURSE: The echocardiographic and angiographic results indicated a dilated cardiomyopathy. Competitive sports activities were stopped and treatment with a beta-blocker (metoprolol) and an ACE-antagonist (ramipril) was started. CONCLUSION: In young male and female athletes, the possibility of severe cardiac abnormalities have to be considered even in the presence of good physical fitness and performance. To reach a high sensitivity for diagnostic ergometry, the work-load must reach the maximal capacity of the cardio-pulmonary system. Differences in the exercise performance of athletes and untrained subjects have to be considered. PMID- 12589584 TI - [Evidence-based treatment of methotrexate-resistant rheumatoid arthritis. Combinations of conventional disease-modifying antirheumatic agents]. PMID- 12589585 TI - [Indications for renal angiography and for percutaneous transluminal renal artery dilatation: interdisciplinary consensus statement regarding renal artery stenosis]. PMID- 12589586 TI - [Private fees of a contract physician in relation to the insured patient. Decision of the Federal Civil Court, 5/172001, B 6 KA 8/00 R]. PMID- 12589587 TI - [How should chronic recurrent cystitis be treated?]. PMID- 12589588 TI - [What is osteopathic medicine?]. PMID- 12589589 TI - [PEG installation in patients unable to give informed consent]. PMID- 12589590 TI - [Angiotensin 1-antagonists: primary therapeutic option in arterial hypertension, too]. PMID- 12589591 TI - [Acarbose can slow down beginning of type 2 diabetes]. PMID- 12589592 TI - [Transcranial Doppler velocimetry in normotensive puerperal women]. AB - OBJECTIVES: To know transcranial Doppler (TCD) normal values in postpartum normotensive women. To evaluate the effect of different variables on cerebral blood flow velocity. PATIENTS AND METHODS: TCD examination was carried out in 100 postpartum normotensive women. Six intracranial arteries were evaluated. Different variables such as arterial pressure, hematocrit and clinical data related to pregnancy and delivery were recorded. A descriptive analysis of qualitative variables was made and the mean and standard deviation of each quantitative variable was calculated. We compared mean velocities (MV) by each side, and assessed the correlation between cerebral blood flow velocity and arterial pressure, hematocrit and the rest of variables. RESULTS: Mean and standard deviations of MV were: right MCA 73.10 14.33; left MCA 75.68 15.84; right ACA 57.61 12.92; left ACA 60.14 14.35; right PCA 42.77 9.28; left PCA 45.54 9.58. Statically significant higher values of MV in the left hemispheric arteries were found. The other analysed variables did not modify the MV values. CONCLUSIONS: In postpartum normotensive women MV values were higher in the left hemisphere. The knowledge of normal puerperium values for TCD is useful to identify cerebral hemodynamic pathological states such those in postpartum eclampsia and postpartum cerebral angiopathy. PMID- 12589593 TI - [Amyotrophic lateral sclerosis: its relationship with thyroid function and phosphate calcium metabolism]. AB - INTRODUCTION: Amyotrophic lateral sclerosis (ALS) is a progressive degenerative disease involving motor neurons of the spinal cord, brain stem, and motor cortex. Previous studies reported the coexistence of motor neuron disease with thyroid dysfunction and primary hyperparathyroidism. These findings suggested an eventual role of the previously mentioned endocrine disorders and the development of ALS. OBJECTIVE: Identify thyroid dysfunction and changes in mineral metabolism in patients affected by sporadic ALS. PATIENTS AND METHODS: We retrospectively analyzed data from 41 in ward patients from June 1998, to June 2001. All of them were diagnosed with sporadic ALS. The following laboratory test were performed: thyrotropin (TSH), thyroxine (T4), serum calcium, serum phosphate, parathyroid hormone (PTH) and ionic calcium concentrations. RESULTS: None of the studied patients was found to be hyperthyroid. Two patients had elevated TSH and normal T4, these findings suggested subclinical hypothyroidism. Elevated PTH was found in one patient; another had primary hyperparathyroidism. Low serum calcium was found in three patients and elevated phosphate was found in another four. CONCLUSIONS: All these findings do not allow us to assert a pathogenic association between thyroid dysfunction or alteration of phosphate calcium metabolism and ALS. PMID- 12589594 TI - [Traumatic brain injury in paediatrics. Our results]. AB - INTRODUCTION: 75% of all children admitted to hospital with traumatic injuries involve traumatic brain injuries (TBI), 70% of the deaths occur within the first 48 hours and mortality fluctuates between 20 and 35%. It is estimated that every year 600,000 consultations are made for this reason, 300 out of every 100,000 children suffer a TBI and about 250,000 cases involve hospitalisation. PATIENTS AND METHODS: A descriptive observational study was carried out of all the patients of paediatric age who attended our Casualty Department in the period between January and December 2001, and had suffered a TBI in any of its clinical variations. The Glasgow coma scale (GCS) was used to classify cases according to their seriousness and results were defined by employing the Glasgow results scale (GRS). RESULTS: A survey of 339 cases with TBI was performed, of which 329 (97%) were mild, eight (2.4%) moderate and two (0.6%) were serious. Of the first, 122 (36%) required hospitalisation and the remaining 217 (64%) were given ambulatory treatment (all of them had mild TBI). Complementary examinations carried out were simple head X rays in 117 (95.9%). CAT (computerised axial tomography) scans were only performed in three cases (2.5%). Only two (1.8%) deaths occurred in our series and there were no cases of disability, according to the GRS. CONCLUSIONS: Mild TBI was the common event in our series. Application of treatment systems that are suited to the conditions in each medical centre helps to reduce the number of hospital admissions for mild TBI. PMID- 12589595 TI - [Antiphospholipid antibodies in patients with vestibular manifestations. A study of 16 cases]. AB - INTRODUCTION: Antiphospholipid antibodies lupus anticoagulant (LA) and anticardiolipin antibodies (aCL) play a role in promoting arterial and venous thrombosis in several vascular territories. Acute vestibular syndromes are a common complaint in general and neurology practice. Approximately 9% of cases are due to central nervous system vestibular areas lesions, often associated with vascular disorders. OBJECTIVE: Define the potential relationship between these antibodies and central or peripheral vestibular failure. PATIENTS AND METHODS: We report the presence of antiphospholipid antibodies in 16 patients with central vestibular symptoms. All patients were seen in the Neuro otology and Vascular Neurology clinics at the Institute for Neurological Research in Buenos Aires. Magnetic resonance imaging (MRI) and ancillary neuro otologic tests were used to determine the etiology of vestibular manifestations. Determinations of LA and aCL were done using standard criteria. RESULTS: We evaluated 16 patients (13 women and 3 men), aged 44 4 years (21 65). Thirteen patients did not have stroke risk factors. MRI lesions were found in 11 subjects (1 cerebellar infarct, 3 pontine ischemic changes, and 9 white matter abnormalities). All patients had signs consistent with dysfunction of vestibulo cerebellar structures or the vestibular nuclei. All patients had positive LA and 4 of them had also elevated aCL. CONCLUSION: Our findings suggest a potential association between the presence of a prothrombotic state and central vestibular dysfunction of vascular etiology. To the best of our knowledge, this is the first report of such an association in the absence of clinically evident autoimmune disease. PMID- 12589597 TI - [Acute infantile bilateral thalamic necrosis]. AB - INTRODUCTION: Acute necrotizing encephalitis is a clinical entity recently described in previously healthy Japanese children. Following a respiratory or gastrointestinal infection of a viral nature, these patients present a sudden deterioration in the level of consciousness. Since neuroimaging shows a bilateral thalamic disorder in all cases, the name infantile bilateral thalamic necrosis has been put forward to refer to this affection. CASE REPORT: We report the case of an 11 month old Spanish patient who was brought into the hospital with disconnection from the surroundings and hypertonia in the upper extremities, within the context of a febrile gastrointestinal picture. Blood sample analysis revealed haemoconcentration and metabolic acidosis, although it did not suggest an infection. The computerised tomography scan of the brain showed a striking hypodensity of both thalami. The initial hypertonia was followed by a generalised hypotonia, accompanied by haemodynamic and respiratory deterioration. The patient died 40 hours after admission. CONCLUSIONS: Acute necrotizing encephalitis was initially described in Japanese children but in recent years isolated cases have also been reported in the Western world. In the case of the patient we have described, one of the most outstanding points is that evolution was unfavourable in spite of mechanical ventilation and the use of inotropic drugs, together with the clinical state of brain death that occurred on the second day after admission. Since neuroimaging showed a selective disorder of the thalami in all cases, and the fact it is an entity that affects small children, it seems more appropriate to use the term acute infantile thalamic necrosis PMID- 12589596 TI - [Sensory demyelinating polyneuropathology as a manifestation of paraneoplastic encephalomyelitis associated to adenocarcinoma of the prostate]. AB - INTRODUCTION: Different neurological disorders affecting different levels of the nervous system, both central and peripheral, are included among the paraneoplastic symptomologies of cancers, although their incidence is not well defined and varies according to the type of tumour. CASE REPORT: We report the case of a 73 year old male patient who was admitted with a suspected polyneuropathy that had been developing for one month. The diagnosis was confirmed by electromyography. The only probable aetiology found in the course of a specific exploration was an adenocarcinoma of the prostate, with a widespread distribution throughout the central ganglionic chains and bone metastases, which were observed by computerised tomography of the abdomino pelvic region and bone gammography, and had not previously been diagnosed. CONCLUSIONS: We reviewed the literature on the relation between neurological paraneoplastic syndromes and neoplasias that are not of a pulmonary origin and we found a low rate of association between the two processes. The relation between adenocarcinoma of the prostate and neurological paraneoplastic processes receives very little attention. These syndromes vary greatly in their neurological expression, and there is a link with different antibodies that could explain an etiopathogenic mechanism of an immunitary nature. No effective treatment exists in spite of its usually being aggressive and varied. The disease often progresses quickly and leads to death after a few months. PMID- 12589598 TI - [T-type primary lymphoma of the central nervous system in immunocompetent patients]. AB - INTRODUCTION: The primary lymphoma of the central nervous system is an infrequent neoplasia, which represents 1,5% of all primary neoplasias in adult patients. In the last decade its frequency has increased threefold, both in immunodepressed as well as in immunocompetent patients. The non Hodgkin lymphoma of B cells being the most frequent histological type, the primary T cell lymphoma of the CNS is a rare clinical entity. CASE REPORTS: In this study we present three cases of immunocompetent patients with primary lymphoma of the central nervous system of T cells seen during the 6 last years in our hospital, the diagnostic imaging by computerized tomography and magnetic resonance showed the tumorations, but the definitive diagnosis was by stereotaxic cerebral biopsy. CONCLUSIONS: The lymphomas are radiosensitive to radiotherapy with survivals of approximately 26 months, the combined treatment of surgery and chemotherapy, prior to radiotherapy, may increases survival up to 48 months. Certain aspects of the patient or of the tumor itself are determining factors with respect to the prognosis of survival. We review the relevant literature and study the clinical manifestation, their value of imaging techniques and differential diagnostic and prognosis of survival PMID- 12589599 TI - [Paraneoplastic ataxia associated to anti CV2 antibodies]. AB - INTRODUCTION: Anti CV2 antibodies are a type of paraneoplastic antibodies that interact with the cytoplasmatic antigens of a subpopulation of oligodendrocytes. They are usually associated with the presence of a small cell carcinoma. We report the case of a patient with an epidermoid carcinoma of the lung and paraneoplastic cerebellous ataxia associated with anti CV2 antibodies. CASE REPORT: Male aged 73, ex smoker for 10 years, with a 45 packet/year habit. He presented a 5 month old constitutional syndrome. Altered gait with a sensation of instability. During the course of the exploration, we detected an increase in the support base, the impossibility to walk in tandem, non exhaustible horizontal nystagmus in bilateral extreme look, and vertical when looking down, with abolition of Achilles reflexes. Brain MR revealed supratentorial demyelinating lesions of a probable ischemic origin. Anti CV 2 antibodies in serum positive (anti HU negative). Biopsy performed using bronchial brushing in the left upper lobe detected the presence of malign epithelial strain, not microcytic, cells, which are compatible with epidermoid carcinoma. DISCUSSION: Anti CV2 antibodies have only been found in patients who have developed a neurological syndrome within the context of a neoplasia. The detection of these antibodies in serum is related with the presence of a concealed neoplasia PMID- 12589600 TI - [Osteochondroma of the atlantooccipital joint. Extreme lateral transcondylar approach. A case description]. AB - INTRODUCTION: We describe a patient diagnosed as suffering from a skull base osteochondroma which affected the atlantooccipital joint and originated in the occipital condyle. It also displayed a growth toward the foramen magnum, which was resected using an extreme lateral transcondylar approach. CASE REPORT: Patient aged 35, with a one year history of vertiginous seizures and unsteady gait, associated with cervical pain that irradiated to the right upper extremity, dysphagia, changes in the tone of the voice and distal numbness of the four extremities. The patient was made to lie in the three quarter prone position and an incision was made in the skin from the C3 spinous apophysis to a point 2 cm below the end of the mastoid process, in relation with the transversal apophysis of C1. The intervention continued with early identification and rotation of the vertebral artery; total resection of the tumour (osteochondroma) with its base in the right occipital condyle and growth toward the foramen magnum, in which the greater resection of the posteromedial third of the condyle is included. CONCLUSIONS: The location of osteochondromas can vary widely, and the condyle is one of the least frequent places inside the occipital bone. The approach employed provides excellent access to the region, in particular to the atlantooccipital joint. The width and angle of exposition are increased as compared with the traditional suboccipital approach, which facilitates the radical resection of the lesion with no neural retraction and without any surgical complications. PMID- 12589601 TI - [New information pathways in the nervous system: communication between astrocytes and neurones]. AB - INTRODUCTION AND METHOD: Astrocytes, a type of glial cell in the central nervous system (CNS), have been classically considered as trophic, structural and supportive cells for neurons. However, in recent years, accumulating evidence suggest a more active role of astrocytes in the physiology of neurons, being involved in the information processing of the CNS. Astrocytes exhibit both a form of excitability based on variations of the intracellular Ca2+ concentration, and a form of communication based on intercellular Ca2+ waves. Furthermore, synaptically released neurotransmitters mobilize Ca2+ from the astrocytic intracellular stores, i.e., the astrocytic cellular excitability can be triggered by the synaptic activity. Finally, astrocytes release the transmitter glutamate to the extracellular space through a Ca2+ dependent mechanism, modulating the neuronal electrical activity and the synaptic transmission. As a consequence of the demonstration of these new forms of cellular communication between astrocytes and neurons, the concept of tripartite synapse has been proposed, in which the synapse is functionally constituted by three elements, i.e., the pre and postsynaptic elements and the surrounding astrocytes. CONCLUSION: The novel results discussed in the present review support the presence of new and complex information pathways in the CNS, which are based on the existence of bidirectional communication between astrocytes and neurons, and which have relevant consequences on the cellular mechanisms responsible for the information processing of the CNS. PMID- 12589603 TI - [Iberoamerican Academy of Pediatric Neurology: its history and importance in the development of Iberoamerican child neurology]. AB - The Iberoamerican Academy of Pediatric Neurology (AINP) started in 1992, in Avila (Spain) and was foundated by a group of pediatrics neurologists who organized three post graduate courses in pediatrics neurology, Miami, Margarita and Cancun, with excelents results, because these reason the AINP has growing, fasted its dramatic success has been mostly due to that group of people who love and build unconditionally with their only goal being the satisfaction of knowing that children with neurological disorders will eventually benefit from their work. In this article we wrote about the history of AINP since 1992, and the reason to foundated. The AINP has growing fasted, and we think that it will be very important in the development of pediatrics neurology in Iberoamerica in different ways like neurology education and the training of the young physician. PMID- 12589602 TI - [Clinical evaluation of multiple sclerosis: quantification by use of scales]. AB - INTRODUCTION: Clinical evaluation is indispensable in multiple sclerosis (MS) for the quantitative measurement of the extent of the disorder, which is in turn required to find out how the disease is evolving and the influence the different forms of treatment are having on it, both in the experimental phase and in the usual monitoring they are subjected to. METHOD: We review the different scales that are used to evaluate the distinct symptomatic and functional aspects of MS and the repercussions these have on the extent of disability displayed in the patient s social and personal life. Although in recent years, the EDSS has been an essential, irreplaceable scale in MS, other instruments of measurement that complement it have also begun to appear. The fatigue, cognitive function and quality of life scales are being used more and more frequently. The Multiple Sclerosis Functional Composite is an instrument that is used more and more frequently in MS and has proved to be highly sensitive in the evaluation of very important clinical trials. CONCLUSIONS: The lack of correlation between the distinct scales corroborates the fact that they measure complementary aspects of MS. PMID- 12589604 TI - [Cerebral palsy: new pathogenetic concepts]. AB - OBJECTIVE: To review the current knowledge pertaining to the new pathogenetic aspects of cerebral palsy (CP). DEVELOPMENT: CP is a group of static, heterogeneous clinical syndromes, characterized by abnormal postural mechanisms and motor activities. Its prevalence in industrialized countries is 2 2.5/1000 newborns. CP should be differentiated from certain genetic or metabolic conditions with which it can be mistaken. Some cases of CP have a genetic basis and they are inherited following a mendelian pattern or are determined by specific isolated genes. Recently, the elevation of certain coagulation factors (i.e., Leiden factor V mutation) and cytokines (i.e. interleukins, a tumor necrosis factor) and interferons have been related to CP pathogenesis. Hypocapnia with PaCO2< 35 mmHg represents a risk for periventricular leukomalacia (PVL) in premature infants. PVL pathogenesis is complex and includes a series of mechanisms that interact among them: fetal/maternal infection, immuneinflammatory reaction, prematurity, intraventricular hemorrhage/iron, ischemia/reperfusion, free radical production, maturational sensitivity of oligodendrocytes, and glutamate effect. Neuroradiological and neuropathological data have demonstrated a cortical anatomical substrate for the intellectual deficits associated with PVL in premature infants. CONCLUSIONS: Current knowledge about CP pathogenesis opens the possibility of early diagnosis and development of new treatments, both therapeutic and preventive. PMID- 12589605 TI - [Management of children with autism spectrum disorders]. AB - INTRODUCTION: Autistic spectrum disorders (ASD) are characterized by qualitative deficits in communication, social skills and a restrictive repertoire of interests and behaviors. Recent studies suggest that they may represent one of the most prevalent neurodevelopmental disorders. METHOD: Treatment of core deficits of ASD are primarily speech and language, behavioral and educational interventions. Clinical management of ASD requires a multidisciplinary approach to identify behaviors that could interfere with the individual s ability to benefit from those interventions. Medications, such as neuroleptics, antiepileptic drugs, SSRI s, neurostimulants and others have been used to target specific behaviors. CONCLUSIONS: Further research is much needed if medications are eventually to have a greater role in the treatment of core ASD symptoms. Recent attempts at neurosurgical interventions cannot be justified by our present level of knowledge PMID- 12589606 TI - [The attention deficit hyperactivity disorder myth or reality?]. AB - INTRODUCTION: The attention deficit hyperactivity disorder (ADHD) has been defined as a disorder of self control characterized clinically by a short attention span, excessive motor activity and poor impulse control. OBJECTIVE: The main objective is to describe how methodological difficulties related to the design of epidemiological studies make an important contribution to the group of heterogeneous data which might lead to doubts regarding the validity of ADHD in children. In recent decades the psychiatric definition of this disorder has varied. This has affected the number and combination of clinical signs necessary for diagnosis of ADHD. The variation in the prevalence rates of the disorder reported in different studies is largely due to these changes. Other factors involved are related to the assessment methods, the type of sample (clinical or community), the source of information (parents, teachers and/or children) and sociocultural characteristics. CONCLUSIONS: ADHD is a valid disorder. It cannot be considered to be a myth since there is considerable evidence of its existence and clinical characteristics in children in different countries. PMID- 12589607 TI - [Cerebral malformation in the newborn: holoprosencephaly and agenesis of the corpus callosum]. AB - OBJECTIVE: To review the embriology and clinical aspects of holoprosencephaly and agenesis of the corpus callosum. DEVELOPMENT: The rostral neuropore closes at 24 days of gestation. At 74 days of gestations axons cross through the dorsal region of the commissural plaque and start forming the corpus callosum. At 115 days of gestations the corpus callosum reaches its adult form. Holoprosencephaly occurs due to rostral mesodermal dysfunction. The etiology of holoprosencephaly is heterogenous. The risk of holoprosencephaly of neonates born to mother with gestational diabetes is 1 2 %. Agenesis of the corpus callosum not associated to holoprosencephaly is due to a disorder of telencephalic midline. Pyruvate dehydrogenase and non ketotic hyperglycinemia can produce agenesis corpus callosum. The prognosis of patients with agenesis of the corpus callosum is dictated by the associated central nervous system and none central nervous system anomalies. CONCLUSIONS: Knowledge of the embriology of holoprosencephaly and agenesis of the corpus callosum are necessary to understand their radiological appearance. The evaluation of a neonate with these conditions requires genetic and metabolic evaluations. PMID- 12589608 TI - [Prevention of mental retardation]. AB - OBJECTIVE: To present within the general field of the conditions causing mental retardation, the preventive strategies for specific application available at the present time. DEVELOPMENT: In spite of the fact that in the majority of cases of mental retardation the etiology is unknown, and for that reason, in them it is not possible to establish preventive strategies, within the last three decades, important research advances have helped to prevent thousands of cases of mental retardation of illnesses caused by Haemophilus influenzae B, measles encephalitis, Rh disease and severe jaundice in newborn infants, congenital hypothyroidism, phenylketonuria and congenital rubella; as well as removing lead from the environment, intervention programs for the proper use of seat belts, child safety seats, and motorcycle and bicycle helmets; early and adequate prenatal care, dietary supplementation with folic acid beginning before conception to reduce the risk of neural tube defects, avoidance of toxic substances during pregnancy like alcohol, and the use of newborn screening tests. CONCLUSION: The primary and secondary prevention of conditions that cause mental retardation continue being a challenge. Require of a review of the present strategies, that frequently inform about the problem, but are not practice in an every day bases (ej. intake of alcohol during pregnancy, the universal use of seat belt and child safety seats during automobile travel). In the future we may have the possibility of prenatal gene therapy. PMID- 12589609 TI - [Acute cerebellar atrophy caused by diphenylhydantoin intoxication resulting from drug interaction]. PMID- 12589610 TI - [Atypical hemicrania as a presenting symptom of a sentinel headache]. PMID- 12589611 TI - [Ependymoma that conditions extended cervical/thoracic/lumbar syringomyelia. A case report and review of the literature]. PMID- 12589613 TI - Effect of a hip flexor-stretching program on gait in the elderly. AB - OBJECTIVES: To test whether a reduction in peak hip extension during the terminal stance phase of walking in elderly compared with young adult subjects represents a hip flexor contracture impairment rather than some dynamic consequence and to test the hypothesis that stretching the hip flexors improves both static and dynamic peak hip extension, as well as other age-related gait changes about the ankle. DESIGN: A double-blinded, randomized, controlled trial. SETTING: Stretching exercises were performed in the subjects' homes. Pre- and postassessments were performed in a gait laboratory. PARTICIPANTS: Ninety-six healthy elderly individuals in 2 groups: treatment (n=47) and control (n=49). INTERVENTION: The treatment group received a 1-time instruction in hip flexor stretching, whereas the control group received a 1-time instruction in shoulder abductor stretching. Participants in each group were asked to perform stretching exercises on their own twice daily for 10 weeks. MAIN OUTCOME MEASURES: Static and dynamic peak hip extension, peak anterior pelvic tilt, and other peak kinematic and kinetic variables during the gait cycle. RESULTS: There was a modest improvement in static peak hip extension as measured by a goniometer within the treatment group (mean +/- standard deviation, 6.1 degrees +/-2.5 degrees to 7.7 degrees +/-3.6 degrees, P=.032) compared with no change in the control group. At comfortable walking speed, dynamic hip extension tended to increase in the treatment group (5.1 degrees +/-9.7 degrees to 7.1 degrees +/-8.0 degrees, P=.103) compared with no real change in the control group (5.3 degrees +/-8.9 degrees to 5.4 degrees +/-7.5 degrees, P=.928). Similarly, at fast walking speed, dynamic hip extension tended to increase in the treatment group (6.4 degrees +/-9.8 degrees to 8.4 degrees +/-8.0 degrees, P=.093) compared with no change in the control group. Changes in ankle kinematics and kinetics included a significant improvement in peak ankle plantarflexion and a tendency to improved ankle power generation. CONCLUSION: The static and dynamic trends to improvement in peak hip extension were of similar magnitude, suggesting that age-related reduction in peak hip extension during gait is the result of a static hip flexion contracture rather than a dynamic consequence. Additionally, age-related changes in ankle kinematics and kinetics may be secondarily related to hip flexion contracture impairment rather than impairment at the ankle per se. This study was limited by the exercises being unsupervised and relying on 1-time instruction. A more rigorous and supervised hip flexor-stretching exercise program may yield more substantial improvements in gait parameters. PMID- 12589614 TI - Toe walking: muscular demands at the ankle and knee. AB - OBJECTIVE: To compare the relationship between electromyographic activity and internal moment in heel-toe and toe walking. DESIGN: Simultaneous recording of stride characteristics and kinematic, kinetic, and intramuscular electromyographic data; paired t tests identified significant between-condition differences. SETTING: Gait laboratory. PARTICIPANTS: Ten able-bodied subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Kinematic, moment, power, and electromyographic variables (ankle, knee). RESULTS: Compared with heel-toe walking, toe walking showed greater plantarflexion during stance (P<.001), higher plantarflexor moments (peak, mean) during loading response (P<.001) and midstance (P<.001), lower mean plantarflexor moments during terminal stance (P=.002), premature soleus (P=.001) and gastrocnemius (P<.001) activity, and higher levels of mean soleus and gastrocnemius activity during stance. During toe walking, the peak internal knee extensor moment was lower in midstance (P=.002), and power absorption was reduced in loading response; however, vastus intermedius electromyographic activity was not reduced. CONCLUSIONS: During toe walking, terminal stance soleus and gastrocnemius activity was greater, despite a lower mean internal plantarflexor moment. The dichotomy between internal moments and muscle effort (ie, electromyographic activity) was consistent with the reduction in force-generation capacity of the calf muscles when the ankle was in a plantarflexed position. PMID- 12589615 TI - The relationship between anthropometric factors and body-balancing movements in postural balance. AB - OBJECTIVE: To evaluate the relationship between anthropometric characteristics and body-balancing movements when standing on 2 legs with eyes open and eyes closed. DESIGN: Cross-sectional. SETTING: A university physiatry laboratory. PARTICIPANTS: One hundred randomly selected subjects (50 men, 50 women; age range, 31-80y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Anthropomorphic: body height, weight, lower-extremity distances, foot lengths, and widest widths of the forefeet and heels. Body movements: 2-legged stance with eyes open and eyes closed, measured with the MacReflex Motion Capture System and calculated as maximal and total movements in 3 dimensions. The relation between the measured anthropometric characteristics and body-balancing movements was calculated by using regression analysis. RESULTS: In the eyes-open condition, maximal lateral knee movement was related to body height and foot length (R(2)=.065, P<.05). Both anteroposterior (AP) head movement (R(2)=.068, P<.05) and AP navel movement (R(2)=.083, P<.05) were related to heel width. AP knee movement was related to foot length and heel width (R(2)=.089, P<.05). Body mass index was related to AP ankle movement (R(2)=.074, P<.05) and to vertical ankle movement (R(2)=.063, P<.05). In the eyes-closed condition, body mass index was related to the vertical navel movement (R(2)=.059, P<.05) and body height to AP knee movement (R(2)=.041, P<.05). CONCLUSION: The levels of significance are not high but warrant attention. It seems that there was no single anthropometric factor that explained the variations in body-balancing movements during standing. PMID- 12589616 TI - Treatment of chronic lumbar diskogenic pain with intradiskal electrothermal therapy: a prospective outcome study. AB - OBJECTIVE: To determine the clinical efficacy of intradiskal electrothermal annuloplasty in treating patients with chronic constant lumbar diskogenic pain who have not responded to at least 6 months of aggressive nonoperative care. DESIGN: Prospective case series. SETTING: Academic-affiliated private physiatry practice. PARTICIPANTS: Thirty-three patients with chronic constant lumbar diskogenic pain of more than 6 months in duration diagnosed with history and physical examination, with concordant pain on provocative pressure-controlled lumbar diskography, and with symptomatic annular tears and/or protrusions less than 5mm, who did not respond to aggressive nonoperative care. INTERVENTION: Intradiskal electrothermal annuloplasty. MAIN OUTCOME MEASURES: Visual analog scale (VAS) pain scores for the back and for the lower extremity, the Roland Morris Disability Questionnaire (RMDQ), and the North American Spine Society Patient Satisfaction Index. RESULTS: A total of 33 patients, with mean age of 40 years and a mean duration of symptoms of 46 months, were observed with a mean follow-up of 15 months. Relief of pain and improvement in physical function were associated with a mean change in the VAS score of 3.9 (P<.001), a mean change in the lower-extremity VAS score of 3.7 (P<.001), and a mean change in the RMDQ of 7.3 (P<.001). For patient satisfaction, 75.7% reported that they would undergo the same procedure for the same outcome. Complete pain relief was achieved in 24% of the patients, and partial pain relief in 46% of the patients. CONCLUSIONS: Intradiskal electrothermal annuloplasty offers a safe, minimally invasive treatment option for carefully selected patients with chronic lumbar diskogenic pain who have not responded to aggressive nonoperative care. PMID- 12589617 TI - Functional independence staging: conceptual foundation, face validity, and empirical derivation. AB - OBJECTIVE: To develop a staging system for functional independence across the activities of daily living (ADLs), sphincter-management, mobility, and executive function domains (ASME) for the FIM instrument that is consistent with the International Classification of Functioning, Disability and Health. DESIGN: National data were used to define the stages. We searched for the most likely configurations of item scores that increased ability to perform component activities in each domain by approximately 1 level per item per stage. SETTING: Inpatient rehabilitation facilities. PARTICIPANTS: Data from 218,290 people discharged from 560 US inpatient rehabilitation facilities in 1995. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Activity profiles formed from FIM scores. RESULTS: Seven stages were defined separately for each ASME domain. Stages approximate the average amount of effort expended by the patient when performing the component activities included in a domain, beginning with less than 25% of effort at the lowest total assistance (stage 1) and ending with 100% effort at the highest complete independence (stage 7). Consistent with developmental principles, independence is achieved at lower stages in the most fundamental activities of eating, transfers, and communication. Recovery of independence in the more difficult activities of bathing, stair climbing, and problem solving does not occur until the higher stages are reached. The degree of independence is described with a shorthand abbreviation of the domains followed by the stage the patient has reached in each domain. For example, ASME 5,1,6,7 indicates need for supervision in the ADLs (A-5), total assistance in sphincter management (S-1), modified independence in mobility (M-6), and complete independence in executive functions (E-7). CONCLUSIONS: ASME stages serve as a common language and shorthand for expressing the functional consequences of illness and injury, while complementing information about impairment and diagnosis, thereby facilitating communication, assessment, and goal setting in terms that are meaningful to patients and their care givers. PMID- 12589618 TI - Staging functional independence validity and applications. AB - OBJECTIVES: To establish construct and predictive validity of the activities of daily living, sphincter-management, mobility, and executive-function (ASME) staging system of functional independence and to describe potential applications. DESIGN: National data were used to validate the stages. Sensitivity to functional change was addressed by comparing patients' stages at discharge to their stages at admission. Construct validity was assessed by examining stage distribution differences among groups of patients classified by impairment category. Predictive validity was determined by the degree to which high stages at admission predict discharge to the community and low stages at admission predict serious morbidity or mortality. SETTING: Inpatient rehabilitation facilities. PARTICIPANTS: Data from 231,686 individuals discharged from 560 US inpatient rehabilitation facilities in 1995. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Stage at rehabilitation discharge, community discharge, acute hospital discharge, or death while in rehabilitation. RESULTS: More than 90% of patients were discharged at a higher stage in at least 1 domain while not declining in any other domain. The odds (95% confidence interval) of community discharge for patients admitted at the highest compared with the lowest stage varied from 2.1 (1.9-2.2) to 3.1 (2.9-3.2), depending on the domain. The odds of acute hospitalization or death were reduced at the highest stages, compared with the lowest, by a factor ranging from .33 (.22-.49) to .65 (.61-.69). CONCLUSIONS: The ASME stages appear valid and sensitive to change. Functional Independence Staging (FIS) has relevance in establishing prognoses and in selecting among therapeutic modalities. FIS will facilitate the identification of meaningful functional goals and interpretable clinical trial endpoints. One of its most important applications could be in the establishment of outcome indicators for quality monitoring under the prospective payment system. PMID- 12589619 TI - The effect of inaccurate FIM instrument ratings on prospective payment: a study of clinician expertise and FIM rating difficulty as contributing to inaccuracy. AB - OBJECTIVE: To test whether clinician expertise and FIM instrument rating difficulty explain clinician overconfidence in FIM rating accuracy. DESIGN: Participants answered 60 true/false FIM questions and, for each question, completed a 6-category scale to assess confidence in the accuracy of their responses. Experts and novices, as well as hard and easy items, were identified through a Rasch analysis. The relation between confidence and accuracy was examined for these different groups. SETTING: Three urban medical centers. PARTICIPANTS: Fifty medical rehabilitation professionals, including physical therapists, occupational therapists, rehabilitation psychologists, speech pathologists, and rehabilitation nurses. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Observed proportion of correct responses to 60 true/false questions and responses from the 6-category confidence scale. RESULTS: The amount of overconfidence was mediated by the difficulty of the FIM task and the level of expertise of the clinical judge. CONCLUSIONS: Decreasing the level of overconfidence in FIM scoring is a promising avenue for improving the accuracy of functional assessment. Accurate assessment of functional status for case-mix group classification will be of even greater importance under the recently initiated Medicare prospective payment system. PMID- 12589620 TI - Behavioral assessment of unilateral neglect: study of the psychometric properties of the Catherine Bergego Scale. AB - OBJECTIVE: To assess the psychometric properties of a scale for spatial neglect in everyday life. DESIGN: Validation study. SETTING: A neurologic rehabilitation unit in a university hospital. PARTICIPANTS: Eighty-three consecutive right hemisphere stroke patients. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The Catherine Bergego Scale (CBS) was used to assess neglect behavior and anosognosia. Its sensitivity was compared with that of 3 conventional tests. The inner structure of the scale was studied by principal component analysis. In addition, linearity, unidimensionality, and reliability of the scale were tested through Rasch analysis. RESULTS: Behavioral assessment correlated significantly to, but was more sensitive than, conventional tests. Anosognosia correlated significantly with neglect severity, although individual dissociations occurred. Factorial analysis disclosed a single underlying factor, explaining 65.8% of total variance. Rasch analysis also revealed that the 10 items defined a common, single ability continuum with widespread measurement range and quite regular item distribution, and showed a satisfactory reliability. CONCLUSION: Behavioral assessment proved to be more sensitive than conventional paper and pencil tasks. Both conventional statistics and Rasch analysis suggest that the CBS is reliable and valid, and that the 10 items define a homogeneous construct. PMID- 12589621 TI - The effects of dehydration on rehabilitation outcomes of elderly orthopedic patients. AB - OBJECTIVE: To study the effects of dehydration, by using the indices of prerenal azotemia and orthostasis, on the rehabilitation outcomes of elderly orthopedic patients. DESIGN: Prospective, pilot study. SETTING: Regional inpatient rehabilitation center. PARTICIPANTS: A consecutive sample of 39 patients (29 women, 10 men), ranging in age from 58 to 94 years (mean, 78y), of whom 13 had total hip replacements, 12 had total knee replacements, and 14 had hip fractures. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Length of stay (LOS), change in the FIM trade mark instrument score, and discharge to home. RESULTS: The mean LOS was significantly longer in the group with prerenal azotemia (n=21, 12.9+/-3.0d, t=2.49, P<.01) than in the nonazotemic group (n=18, 9.4+/-4.6d). LOS was also significantly longer in the orthostatic group (n=18, 13.7+/-3.4d, t=2.94, P<.01) than in the nonorthostatic group (n=21, 9.8+/-3.9d). Two-way analysis of variance showed a statistically significant effect on LOS for both azotemia (F=8.4, P=.006) and orthostasis (F=10.5, P=.003). A statistical interaction existed (F=4.7, P=.038), but it was more pronounced in the absence of both conditions, as opposed to the presence of both. LOS for the group with both azotemia and orthostasis was 13.6+/-2.7 days (n=10, F=4.7, P=.038), in contrast to 7.2+/-2.8 days in the group without either condition (n=10). Of the patients who had neither azotemia nor orthostasis, 100% (n=10) went home; 80% (n=8) of patients who had both conditions went home. Logistic regression analysis, however, did not show a statistically significant correlation between discharge to home and the presence of azotemia, orthostasis, or both. CONCLUSION: Prerenal azotemia and orthostasis are present in a significant number of elderly orthopedic patients and have a major effect on rehabilitation outcomes. PMID- 12589622 TI - Work status after distal femoral Kotz reconstruction for malignant tumors of bone. AB - OBJECTIVE: To determine the proportion of patients who experienced altered work status after reconstruction with a modular tumor prosthesis and adjuvant chemotherapy for primary sarcoma of the distal femur. DESIGN: We reviewed an institution's tumor database and patients' charts. Disability and work status data were collected preoperatively, at 1 year and 2 years postoperatively and at final follow-up. SETTING: Acute-care teaching hospital in Ontario, Canada. PARTICIPANTS: Thirty-six eligible patients. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Work status was plotted over time to identify unexpected deviations (based on age and preoperative status), and subjects were assigned to altered work status (defined by detrimental changes in productivity, eg, change from full-time work to not working) or expected work status (defined by ongoing work or students progressing to higher education). Disability was measured with the Toronto Extremity Salvage Score and the Musculoskeletal Tumor Society score. RESULTS: Nine patients (25%) experienced altered work status. There were no significant differences in disability ratings or demographic information between the 2 groups. Baseline job demand and living environment (rural vs urban, P=.001) were predictive by logistic regression. CONCLUSIONS: Patients with physically demanding jobs, and those living rurally, are at greatest risk for altered work status. PMID- 12589623 TI - The pendulum test for evaluating spasticity of the elbow joint. AB - OBJECTIVE: To develop a modification of the pendulum test to allow evaluation of elbow spasticity. DESIGN: The main difficulties of directly applying the conventional pendulum test to the elbow were the small inertia of the forearm and the uncomfortable posture. We designed an experimental apparatus similar to a clock pendulum and developed an elbow biomechanic model to measure objectively spasticity of the elbow joint. The model consisted of linear stiffness and damping and gravity contribution. SETTING: A referral medical center in Taiwan. PARTICIPANTS: Eleven stable stroke patients and 11 able-bodied subjects. INTERVENTIONS: A custom-designed accessory apparatus to facilitate the pendulum test in elbow joints. MAIN OUTCOME MEASURES: By using an optimization technique, we estimated parameters of the proposed elbow biomechanic model as the candidate indicators of spasticity. RESULTS: The stiffness constant remained relatively consistent in all groups. Both the damping coefficient and damping ratio increased in the affected side of stroke patients and tended to increase with the severity of spasticity. Damping ratio had marginally better differentiation capability than the damping coefficient. CONCLUSIONS: The damping ratio derived from the proposed model differentiated spasticity from normotonus and increased as spasticity increased. PMID- 12589624 TI - Validation of postural radiographs as a way to measure change in pelvic obliquity. AB - OBJECTIVE: To evaluate the validity of using postural radiographs to assess change in pelvic obliquity. DESIGN: Blinded comparison of reference lines and instrument development of postural radiographs. SETTING: Veterans Administration hospital. PARTICIPANTS: Thirty-nine subjects with no history of low back pain. INTERVENTION: Repeated postural radiographs without heel lifts and with (1/4)-in (6.35-mm) and (1/2)-in (12.7-mm) heel lifts. MAIN OUTCOME MEASURE: The change of pelvic obliquity as measured by the intersulcate, interalar, and sacral surface lines on postural radiographs. RESULTS: A (1/4)-in heel lift resulted in changes of 4.6+/-4.0mm, 3.6+/-3.1mm, and 5.8+/-3.9mm and a (1/2)-in heel lift in changes of 7.8+/-4.3mm, 7.8+/-4.6mm, and 5.2+/-3.8mm for the intersulcate, interalar, and sacral surface lines, respectively. Pearson correlation coefficients were used to correlate the change in pelvic obliquity with the corresponding heights of heel lifts; for the (1/4)-in heel lift, they were.67,.88, and.71, and for (1/2)-in heel lift they were.73,.79, and.87 for the intersulcate, interalar, and sacral surface lines, respectively (all P<.001). CONCLUSION: All the reference lines seem to be valid measures of change of pelvic obliquity relative to the size of the lifts (ie, the changes in pelvic obliquity seem to be causally related and correlated with the lifts). These results also suggest that postural radiographs can be used to determine the magnitude of pelvic obliquity. PMID- 12589625 TI - Intention tremor rated according to different finger-to-nose test protocols: a survey. AB - OBJECTIVES: To investigate the dependence of intention tremor rating scores on different finger-to-nose test (FNT) protocols, varying in arm position and the time the finger has to be kept on the nose, and to examine their relevance to function. DESIGN: Observational survey. Video recordings were made while patients performed the FNTs and functional tasks such as pouring water. SETTING: Three European rehabilitation centers in Belgium, Finland, and England. PARTICIPANTS: Twenty-six multiple sclerosis patients with intention tremor (avg age, 44.1y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Six examiners rated the degree of intention tremor by using the Fahn Tremor Rating Scale. RESULTS: Interrater reliability for rating intention tremor during the FNTs was high (kappa=.65-.74). Both the required arm position and time constraints affected the magnitude of intention tremor rating scores (mean rank, 2.27-2.95) and their functional relevance (rho=.70-.84). Intention tremor was scored the highest when the arm was lifted to 90 degrees of abduction at the shoulder and the subject was required to stabilize the finger on the nose. However, its functional relevance was lower compared with a more "functionally" executed FNT. CONCLUSIONS: The rating of intention tremor during the FNT depends on test instructions. A standardized protocol is needed. PMID- 12589626 TI - Value of sensory examination in predicting bladder function in patients with T12 L1 fractures and spinal cord injury. AB - OBJECTIVE: To determine whether early sensory examination, voluntary anal sphincter contraction, or bulbocavernosus reflex (BCR) might predict bladder function in patients with a spinal fracture at the thoracolumbar level. DESIGN: Longitudinal study of consecutive patients admitted to a spinal cord injury (SCI) rehabilitation center. SETTING: Primary care center, university facility in Switzerland. PARTICIPANTS: Fifty-five patients with thoracolumbar fractures. INTERVENTIONS: Neurologic (American Spinal Injury Association [ASIA] protocol) and urodynamic examination during the first hospitalization and at follow-up. MAIN OUTCOME MEASURES: Neurologic sensory scores and type of neurogenic bladder. RESULTS: At first examination, there was no correlation between the sensory examination, voluntary anal sphincter contraction, BCR, and neurogenic bladder type. At follow-up (time since first examination: mean, 698+/-47.2d; median, 481d), the sensory examination remained of no value in distinguishing the neurogenic bladder type. However, voluntary anal sphincter contraction distinguished between complete and incomplete neurogenic bladders and BCR differentiated between complete bladder dysfunction of the lower motoneuron and upper motoneuron type. At follow-up, the bladder function (51 patients) remained unchanged in 44 cases and normalized in only 7 cases. Patients who improved their bladder function tended to have higher initial sensory ASIA scores (P<.05, Kruskal-Wallis test). Of the 7 patients who improved their bladder function, all but 1 (85%) had initial perineal pinprick sensation. Nevertheless, preservation of perineal pinprick sensation was of no positive predictive value, because 21 patients (48%) who initially had perineal pinprick sensation did not improve their voiding function, a finding similar to that of the 23 (52%) without initial perineal pinprick sensation whose bladder function also did not improve. CONCLUSIONS: In SCI patients with thoracolumbar fractures, neurogenic voiding dysfunction cannot be predicted by the sensory evaluation. In patients with an SCI at the thoracolumbar level, pinprick sensation in the perineal area is of negative predictive value: absence of pinprick sensation predicts poor bladder recovery. Most patients with a spinal fracture at T12-L1 did not improve in voiding function. PMID- 12589627 TI - Comparison of the ankle, knee, hip, and trunk corrective action shown during single-leg stance on firm, foam, and multiaxial surfaces. AB - OBJECTIVE: To compare the quantity of ankle, knee, hip, and trunk corrective actions shown during single-leg stance. DESIGN: Counter-balanced crossover design. Single-leg stance under the conditions of eyes open on firm, foam, and multiaxial surfaces and eyes closed on a firm surface were recorded for 12 seconds in 18 participants. SETTING: A university neuromuscular research laboratory. PARTICIPANTS: Eighteen healthy and recreationally active college students. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Average angular displacement magnitude between successive sampling instances for the ankle, knee, hip, and trunk. RESULTS: A significant condition by joint interaction was revealed. Post hoc comparisons revealed that the ankle dominated as the source of corrective action across each of the testing conditions. As the challenge became greater because of foam surface or eyes closed, more corrective action occurred at proximal joints (hip and/or knee). CONCLUSIONS: The ankle is of primary importance during single-leg stance on firm, foam, and multiaxial surfaces, with proximal joints having an increased role under more challenging conditions. These results provide a scientific basis for clinicians' and researchers' decisions about support surface and visual condition during single-leg postural control testing and training. PMID- 12589628 TI - Seat and footrest shocks and vibrations in manual wheelchairs with and without suspension. AB - OBJECTIVE: To examine differences in the shock and vibration transmitted to an occupant of a manual wheelchair with and without suspension caster forks and with and without rear-suspension systems. DESIGN: Repeated-measures engineering testing. SETTING: Rehabilitation engineering center with a wheelchair standards test laboratory. SPECIMENS: Six manual wheelchairs. INTERVENTIONS: An American National Standards Institute/Rehabilitation Engineering and Assistive Technology Society of North America wheelchair test dummy and a Hybrid III test dummy were used to test shock and vibration transmission in wheelchairs equipped with original equipment manufacturer (OEM) caster forks and suspension caster forks. Ultralight wheelchairs, half of which had factory-equipped rear-suspension systems, were tested. Testing was conducted on a double-drum wheelchair test machine. MAIN OUTCOME MEASURES: Shocks were examined by using peak acceleration and the frequency at which peak acceleration occurs for the seat and footrest. Vibration was characterized by the acceleration power per octave for the seat and footrest. RESULTS: Significant differences were found in the peak accelerations at the seat (P=.0004) and footrest (P=.0007) between the wheelchairs with the OEM caster forks and those with the suspension casters. The wheelchairs with suspension had significantly different frequencies at which the peak accelerations occurred for both the seat (P=.01) and footrest (P=.0001). The wheelchairs with suspension caster forks had a lower total power per octave than the wheelchairs with the OEM caster forks. For the footrest vibrations, significant differences were found between the types of caster forks for all octaves except those associated with frequencies more than 78.75Hz. There were significant differences for wheelchairs with and without rear suspension for total power per octave of seat vibrations in the octaves between 7.81 and 9.84Hz (P=.01) and 12.40 and 15.63Hz (P=.008). CONCLUSIONS: Suspension caster forks reduce the shock and vibration exposure to the user of a manual wheelchair. Rear suspension systems reduce some of the factors related to shock and vibration exposure, but they are not clearly superior to traditional designs. PMID- 12589629 TI - Treatment with botulinum toxin type B for upper-limb spasticity. AB - OBJECTIVE: To determine if botulinum toxin type B (MyoBloc) decreases spasticity. DESIGN: Investigator-initiated trial, open-label, single-treatment session. SETTING: Single site. PARTICIPANTS: Ten patients with stable upper-limb spasticity and an Ashworth Scale score of 2 or higher at the elbow, wrist, and fingers. INTERVENTIONS: Total dose of 10,000U of botulinum toxin type B injected into 5 major muscles. MAIN OUTCOME MEASURES: The Ashworth Scale, goniometry, and functional assessments were performed at injection and weeks 4, 8, and 12. The principal investigator (PI) global assessment of change (GAC) and the patient GAC were done at weeks 4, 8, and 12 postinjection. The safety of the procedure was measured by adverse events and vital signs. RESULTS: Improvements in Ashworth Scale scores were observed at weeks 4, 8, and 12 postinjection. At week 4, the mean changes (in Ashworth score) were elbow, -1.0 (P=.016); wrist, -1.7 (P=.004); finger, -1.35 (P=.02); at week 8: elbow, -.83 (P=.016); wrist, -1.00 (P=.016); finger, -.94 (P=.08); and at week 12: elbow, -.61 (P=.07); wrist, -1.00 (P=.016); and finger, -.89 (P=.10). The PI GAC improved at all visits. Nine of the 10 subjects reported dry mouth at week 4, with resolution by week 12. No changes were seen on the functional measures. CONCLUSIONS: Botulinum toxin type B may be useful in treatment of spasticity. PMID- 12589630 TI - Rectal pressure response to a meal in patients with high spinal cord injury. AB - OBJECTIVES: To determine whether there is a postprandial increase of rectal pressure in patients with spinal cord injury (SCI) and to compare their rectal pressures with those of healthy volunteers. DESIGN: A before-after trial comparing SCI and control subjects. SETTING: Patients were recruited from the register of an SCI unit at a rehabilitation medicine department, and the study took place at the university hospital laboratory in Sweden. PARTICIPANTS: Ten patients with high traumatic SCI and 9 healthy volunteers. Eight patients had a lesion above level T5. INTERVENTION: Continuous anorectal manometry was performed. Rectal activity was calculated before and at regular time intervals after a 1000-cal test meal. MAIN OUTCOME MEASURE: Rectal activity measured as area under the pressure curve. RESULTS: There was a significant increase in rectal activity of 46% after 10 minutes in the patients but of 72% after 5 minutes in the volunteers. There was no difference in fasting rectal activity, but patients had a stronger mean rectal postprandial response during 60 minutes compared with volunteers. CONCLUSIONS: These results support the theory that the colonic response to food is preserved in patients with high SCI. PMID- 12589631 TI - Cardiovascular and endocrine responses during the cold pressor test in subjects with cervical spinal cord injuries. AB - OBJECTIVE: To investigate cardiovascular regulation and endocrine responses during the cold pressor test in patients with chronic spinal cord injury (SCI). DESIGN: Experimental and control study. SETTING: University laboratory, department of rehabilitation medicine, in Japan. PARTICIPANTS: Eight quadriplegic subjects with complete spinal cord transection at the C6 to C8 level and 6 age matched healthy subjects. INTERVENTIONS: Cardiovascular and endocrine responses were examined during 2 minutes of control, 3 minutes of ice-water immersion of the foot, followed by a 3-minute recovery. MAIN OUTCOME MEASURES: Blood pressure, heart rate, the Borg 15-point Rating of Perceived Pain Scale, and blood samples for measurement of plasma norepinephrine, epinephrine, plasma renin activity, plasma aldosterone, and arginine vasopressin. RESULTS: The rise in the mean arterial blood pressure during the cold pressor test in patients with SCI (baseline, 81.6+/-3.7mmHg; increased by 30%+/-6.1%) was significantly (P<.05) higher than that in healthy subjects (baseline, 101.2+/-4.5mmHg; increased by 20%+/-4.5%). The SCI subjects had no change in heart rate throughout the test, in contrast to the tachycardia noted in normal subjects. Baseline plasma norepinephrine in SCI subjects (63.0+/-18.3pg/mL) was significantly lower than in normal subjects (162.3+/-19.6pg/mL) and plasma norepinephrine increased significantly during the cold pressor test in both groups. CONCLUSIONS: In the SCI subjects, a reflex sympathetic discharge through the isolated spinal cord results in a more profound rise in mean blood pressure during ice-water immersion. This response was free of inhibitory impulses from supraspinal center and baroreceptor reflexes, either of which might restrain the increase in blood pressure. PMID- 12589632 TI - Persons with C5 or C6 tetraplegia achieve selected functional gains using a neuroprosthesis. AB - OBJECTIVE: To test the efficacy and safety of the NESS Handmaster neuroprosthesis with subjects with C5 or C6 tetraplegia. DESIGN: Interventional, nonrandomized case series. SETTING: Subjects' residence and university research laboratory. PARTICIPANTS: Men, 3 to 17 years after C5 (n=5) and C6 (n=2) spinal cord injury (SCI). INTERVENTION: Subjects practiced with the neuroprosthesis daily to regain grasp, hold, and release ability and to restore selected functions of 1 of the 2 paralyzed hands. Subjects were observed 2 to 3 times weekly for 3 weeks. MAIN OUTCOME MEASURES: Three activities of daily living (ADL) tasks: (1) pick up a telephone, (2) eat food with a fork, and (3) perform 1 individually selected ADL task and 2 grasp, hold, and release tasks (lift a videocassette, lift a 150-g weight). Secondary outcomes were grip strength, electrically induced finger motion, and Fugl-Meyer spherical grasp. Nonparametric data were analyzed with the Wilcoxon signed-rank test, and parametric data (grip strength and finger motion) were analyzed by analysis of variance. All tests were considered significant at P equal to.01. RESULTS: At study completion, all 7 subjects were 100% successful at using the Handmaster in the studied ADL and grasp, hold, and release tasks. Significant improvements occurred in grip strength (from.57+/-.98N at baseline to 16.5+/-4.4N), finger linear motion (from 0.0cm at baseline to 8.4+/-3.2cm), and Fugl-Meyer scores. No safety issues were encountered. Six of 7 subjects rated their overall performance as "excellent." CONCLUSIONS: The Handmaster is a safe, noninvasive neuroprosthesis that improves hand function of selected subjects with C5 or C6 SCI. PMID- 12589633 TI - Short-term recovery of limb muscle strength after acute stroke. AB - OBJECTIVES: To document, by using norm-referenced strength measures, the recovery of limb muscle strength of patients undergoing stroke rehabilitation and to examine the relation between comorbidities and the recovery of strength after stroke. DESIGN: Retrospective analysis of data from a consecutive convenience sample of patients examined clinically between 1994 and 1997. SETTING: Acute inpatient rehabilitation unit. PARTICIPANTS: Fifty patients with stroke who were able to follow commands and were examined during acute rehabilitation by a single examiner (AWA). INTERVENTIONS: Stroke rehabilitation emphasizing early movement, exercise with resistance, and daily functional activities. MAIN OUTCOME MEASURE: The strength at discharge of 7 muscle actions (shoulder abduction, elbow flexion, elbow extension, wrist extension, hip flexion, knee extension, ankle dorsiflexion) measured bilaterally with a hand-held dynamometer and compared with norm-referenced values. RESULTS: Differences in strength between admission and discharge were significant for all muscle actions on the weaker side and for 4 of the 7 muscle actions on the stronger side. At discharge, the bilateral strength of all muscle actions was weaker than predicted by data from healthy individuals of comparable age, sex, and weight (F>17.000, P<.001). Strength did not differ between subjects who did and did not have a previous stroke or comorbidities. CONCLUSIONS: Subjects undergoing inpatient rehabilitation soon after stroke experienced an increase in limb muscle strength bilaterally. This increase was not influenced by previous stroke or comorbidities. PMID- 12589634 TI - Cryotherapy influences joint laxity and position sense of the healthy knee joint. AB - OBJECTIVE: To investigate the influences of cryotherapy on the anteroposterior (AP) laxity and the joint position sense of the knee. DESIGN: Experimental. SETTING: University hospital in Japan. PARTICIPANTS: Twenty healthy volunteers (10 men, 10 women; age range, 21-28y) were analyzed. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: A cooling pad was applied to the knee for 15 minutes under the circulating medium at 4 degrees C. The subject's skin temperature over the anteromedial aspect of the knee was measured during the 15 minutes of cooling and again 15 minutes later. The accuracy of the knee joint position sense was evaluated before and after cooling by using a modified Skinner's method. AP displacement and anterior terminal stiffness (ATS) of the knee were obtained with a KT-2000 trade mark knee arthrometer. RESULTS: Total (AP) laxity and anterior knee laxity decreased by 1.0 and 1.2mm after 15 minutes of cooling (P=.003, P=.017), respectively. ATS and inaccuracy of position sense increased by 21N/mm and 1.7 degrees (P<.001, P=.003), respectively. All parameters had normalized at 15 minutes postcooling. CONCLUSION: Cooling for 15 minutes makes the knee joint stiffer and lessens the sensitivity of the position sense. These findings may be significant and should be taken into account for therapeutic programs that involve exercise immediately after a period of cooling. PMID- 12589635 TI - Detrusor and blood pressure responses to dorsal penile nerve stimulation during hyperreflexic contraction of the bladder in patients with cervical cord injury. AB - OBJECTIVE: To investigate the immediate effect of dorsal penile nerve (DPN) stimulation on detrusor pressure (P(det)) and blood pressure during hyperreflexic contractions of the bladder in patients with cervical spinal cord injury (SCI). DESIGN: Blood pressure and P(det) monitoring during cystometry with and without DPN stimulation. SETTING: Urodynamic laboratory in a university hospital in Korea. PARTICIPANTS: Eight men (age range, 20-55y) with cervical SCI that was incurred from 4 months to 10 years before this study. INTERVENTION: During water cystometry, blood pressure was monitored with an intra-arterial catheter introduced percutaneously into the radial artery and was recorded simultaneously with the P(det). Blood pressure was also measured manually with an electronic blood pressure cuff. Electric stimulation was applied to the DPN by using surface electrodes each time a bladder contraction was detected. Stimulation intensity was twice the threshold of the pudendal-anal reflex. MAIN OUTCOME MEASURES: P(det), systolic blood pressure, and diastolic blood pressure. RESULTS: As P(det) increased, the blood pressure increased in all cases. All the reflex contractions of the bladder were effectively suppressed by DPN stimulation, and as the P(det) decreased during stimulation, radial arterial pressure also decreased immediately and significantly. CONCLUSIONS: DPN stimulation can decrease P(det) and the increased blood pressure associated with it. PMID- 12589636 TI - Pharmacologic treatment of complex regional pain syndrome I: a conceptual framework. AB - Pain may be a leading symptom in complex regional pain syndrome type I (CRPS I) and may hinder functional recovery. In this case, a pharmacotherapeutic approach to pain should be part of the individually tailored interdisciplinary treatment regimen. However, operational criteria for determining which patient may profit from what therapeutic intervention are lacking. This article discusses a conceptual framework in which the rapid progress made in basic pain research may contribute to the clinical management of pain in CRPS I. First, recent insights in the pathophysiologic mechanisms underlying CRPS I are reviewed. CRPS I is considered a neuropathic pain syndrome with a mixed and time-dependent profile of a regional inflammation, sensitization of primary somatosensory afferents (peripheral sensitization), and sensitization of spinal neurons (central sensitization). The dominant mechanisms may vary across individual patients with different time profiles. Second, a model was constructed in which signs and symptoms in an individual patient are related to these mechanisms. Finally, relating the clinical picture to the underlying pathophysiology may help determine the pharmacotherapeutic approach for an individual patient. Pharmacologic options are discussed in this context. The presented framework does not aim to provide an evidence-based treatment algorithm, ready to be used in daily clinical practice; rather it offers a crude, first step toward a mechanism based pharmacotherapy in CRPS I, in an effort to shift from a mainly empirical treatment paradigm toward theory-driven treatment procedures. PMID- 12589638 TI - Public expectations, achievable cervical screening sensitivity, and the standard of practice. PMID- 12589637 TI - Disabling overactivity of the extensor hallucis longus after stroke: clinical expression and efficacy of botulinum toxin type A. AB - OBJECTIVES: To describe the clinical characteristics of persistent extension of the great toe related to overactivity of the extensor hallucis longus (EHL) after a stroke and to explore the efficacy of treatment with botulinum toxin type A. DESIGN: Case series. SETTING: A physical medicine and rehabilitation department in France. PARTICIPANTS: Eleven patients with hemiplegia disabled by overactivity of the EHL were classified according to the different ways in which that overactivity was triggered. Disability was related to pain in 5 patients, to shoe difficulties in 10 patients, and to varus foot in 6 patients. INTERVENTION: Injection of botulinum toxin type A into the EHL. MAIN OUTCOME MEASURES: Efficacy was assessed on day 15 by triggering of pain, shoe difficulties, and varus deformity and by patients' subjective assessment on day 15, at 3 months, and at 6 months. RESULTS: On day 15, EHL overactivity disappeared after 16 of the 18 injections in 10 patients; subjective assessment was very good for pain and shoe difficulties and remained good or very good at 3 months for 8 patients who received 12 injections. CONCLUSION: Botulinum toxin type A is effective in treating disability related to poststroke overactivity of the EHL. PMID- 12589639 TI - Urbanization and the incidence of abnormalities of squamous and glandular epithelium of the cervix. AB - BACKGROUND: The large data bases of the Dutch cervical screening program can be exploited to establish the relation between urbanization and the incidence of abnormalities of the squamous and glandular epithelium, including mild or greater changes of the squamous and glandular epithelium of the cervix. METHODS: Six cytology laboratories in the context of the Dutch cervical screening program screened over 190,000 cervical smears. Urbanization (place of residence) data were derived from postal codes. All smears were coded with the Dutch national coding system, the Dutch national classification system KOPAC, in which squamous abnormalities are coded S4-S9, and glandular cell changes are coded G4-G9. From the scores per 1000 screened women, the relative risk (RR) of living in a large city compared with living in rural areas was calculated. To investigate a trend in incidence in relation to urbanization, the Schaafsma method was used. RESULTS: Of the smears with positive cytology, mild squamous dysplasia (S4) had the highest incidence per 1000 screened women (4.32), and the lowest incidence was found for adenocarcinoma (in situ; G7/G9; RR, 0.07). The RR for urban women ranged from 1.73 for moderate squamous dysplasia (S5) to 7.55 for adenocarcinoma (in situ; G7/G9). For smears with positive cytology for both squamous and glandular abnormalities, the Schaafsma method indicated a significant positive trend. CONCLUSIONS: The incidence of squamous and glandular abnormalities are maximal in women who live in a large city, which, in The Netherlands, is where there also is a population at high risk for human papillomavirus and bacterial vaginosis. PMID- 12589640 TI - Metaanalysis of the accuracy of rapid prescreening relative to full screening of pap smears. AB - BACKGROUND: Efficient quality assurance and improvement measures are essential ingredients in a well organized cytology-based program for cervical carcinoma screening. Various pap smear review procedures, aiming for optimization of accuracy, are described throughout the literature. Evaluation and synthesis of those methods are needed. In a previous study, we pooled data on the diagnostic quality of rapid reviewing (RR) of cervical smears initially reported as normal or unsatisfactory. We now focus on rapid prescreening (RPS) of unreported smears. METHODS: Six published studies on the accuracy of RPS relative to subsequent full screening were pooled using metaanalytic methods. Individual and pooled sensitivity, specificity, and predictive values were assessed using forest plots. Random effect pooling methods were used for interstudy heterogeneity. Variation in sensitivity according to influencing factors was explored by metaregression. RESULTS: The pooled average sensitivity of RPS was 64.9% (95% confidence interval [CI] 50.7-79.1%) for all abnormalities, 72.6% (95% CI 60.6-85.2%) for low-grade lesions or more severe, and 85.7% (95% CI 77.8-93.6%) for high-grade lesions or more severe. The pooled specificity was estimated at 96.8% (CI 95.8-97.8%). The sensitivity increased significantly with duration of screening and decreased with workload. Almost 3% of all abnormal slides were detected only by RPS (2.8%; CI 0.0-5.8%). This is comparable to the proportion of false-negative smears detectable by RR. CONCLUSIONS: Rapid prescreening has a high yield for severe dysplasia and shows diagnostic properties that support its use as a quality control procedure in cytologic laboratories. We showed previously that RR is superior to full reviewing of a 10% random sample of negative slides (10% FR). Because the yield of additional abnormalities found by RR and RPS is comparable, we expect RPS to be more efficient than 10% FR as well. PMID- 12589641 TI - Long microvilli of mesothelioma are conspicuous in pleural effusions processed by Ultrafast Papanicolaou stain. AB - BACKGROUND: The differentiation of malignant epithelial mesothelioma, nonmucinous pulmonary adenocarcinoma, and mesothelial hyperplasia by direct light microscopic examination is challenging. The detection of long microvilli, an ultrastructural feature unique for malignant epithelial mesothelioma, requires immunolabeling of epithelial membrane antigen to be visible by light microscopy. The purpose of this study is to report that this feature is now made conspicuous enough to be detected by light microscopy by a different approach in cytologic processing. METHODS: The study includes 9 cases of mesothelioma, 50 cases of mesothelial hyperplasia, and 50 cases of nonmucinous pulmonary adenocarcinoma. Direct smears were made from the sediments of fresh or refrigerated effusions, air dried, rehydrated with normal saline for 30 seconds to create retraction halos, and processed with the Ultrafast Papanicolaou stain (UFP). RESULTS: Long microvilli in thick bundles as well as short, fine projections radiating from the free surface of the epithelial mesothelioma cells were observed within the retraction halos in pleural effusions processed by UFP. These features were not observed in the UFP-processed hyperplastic mesothelial cells or in the pulmonary nonmucinous adenocarcinoma cells. CONCLUSION: Direct visualization of long microvilli via UFP may aid in the distinction of malignant mesothelioma from mesothelial hyperplasia and adenocarcinoma before immunohistochemical and electron microscopic studies are performed. PMID- 12589642 TI - Cytologic features of the nested variant of urothelial carcinoma: a study of seven cases. AB - BACKGROUND: The nested variant of urothelial carcinoma (NVUC) is a recently described rare variant of urothelial carcinoma with deceptively bland morphologic findings, although it does display aggressive behavior. To the authors' knowledge there is no previous report of cytologic findings in urine specimens from patients with NVUC. METHODS: Urine specimens from patients with histologically confirmed NVUC were reviewed. The specimens evaluated included urine specimens obtained concurrently with or up to 1 month preceding the patients' surgical procedure. All the specimens were analyzed for the presence of cells morphologically similar to the NVUC cells observed in the tissue sections. These cells were evaluated for the following parameters: number of neoplastic cells; cellular arrangement; cell size and shape; cell borders; and cytoplasmic, nuclear, and nucleolar features. RESULTS: Thirteen urine specimens from 7 patients were included in the current study. The authors were able to identify cells that were similar morphologically to the cells present in the nests of NVUC in all cytologic specimens. The neoplastic cells for the most part were medium sized, round or polygonal, with abundant, dense, slightly granular basophilic cytoplasm and well defined cell borders. The nuclear/cytoplasmic ratio was increased, the nuclear membranes had irregular contours, and the nuclei contained coarse chromatin with occasional prominent nucleoli. CONCLUSIONS: The cytologic features of NVUC are subtle but distinct. A primary diagnosis of NVUC in urine specimens is not recommended in view of the subtleness of the findings. Nonetheless, the presence of cells with the features described earlier should warrant a cystoscopic examination with histologic confirmation in a patient with a previous history of NVUC. PMID- 12589643 TI - Cytologic features of round cell liposarcoma: a report on five patients. AB - BACKGROUND: Round cell liposarcoma (RCL) and myxoid liposarcoma are now considered as a single pathologic entity for several reasons. Despite many similarities, both morphologic phenotypes may differ considerably, and RCL may be difficult to recognize as liposarcoma. In addition, few cytologic reports describing features of RCL are available. METHODS: Five patients with RCL for whom cytologic study of the tumor was available were reviewed. Only tumors in which round cell or poorly differentiated, hypercellular areas comprised > 75% of the tumor were considered RCL. RESULTS: Smears were hypercellular and consisted of single, round cells with scarce cytoplasm and naked nuclei. Some cells showed slightly oval rather than round nuclei. In three tumors, the background was vacuolated; and, in two tumors, intracytoplasmic vacuoles were present. Isolated fragments of myxoid stroma containing neoplastic cells were identified in two tumors. Pleomorphism was not a prominent finding. CONCLUSIONS: RCL may show morphologic features that are uncommon for a conventional liposarcoma, leading to an erroneous cytologic interpretation. This variant of liposarcoma must be considered when evaluating round cell neoplasms, mainly in adult patients. The presence of oval cells, vacuoles (intracytoplasmic or extracellular), and myxoid stroma should raise this diagnostic possibility, avoiding confusion with other well-known round cell neoplasms. PMID- 12589644 TI - Cytologic and immunocytochemical findings of anaplastic large cell lymphoma: analysis of ten fine-needle aspiration specimens over a 9-year period. AB - BACKGROUND: Anaplastic large cell lymphoma (ALCL) has raised much controversy in the field of hematolymphoid pathology. Its nature is becoming better characterized with recent advances in molecular genetics. However, to the authors' knowledge, a detailed description of the fine-needle aspiration (FNA) cytology of ALCL is lacking and the application of immunocytochemical study, including immunostaining for anaplastic lymphoma kinase (ALK) protein, to cytology samples has not been studied to date. METHODS: The authors reviewed 10 FNA specimens of ALCL from 8 patients encountered at Pamela Youde Nethersole Eastern Hospital and Queen Mary Hospital in Hong Kong over a 9-year period from early 1993 to the end of 2001. The cytologic and immunocytochemical findings (including ALK protein overexpression) of the specimens were correlated with histologic and immunohistochemical findings of surgical biopsy specimens. RESULTS: Six of the eight patients had ALCL of the common variant, whereas the remaining two patients had ALCL of the small cell variant. FNA specimens of ALCL of the common variant yielded many loosely dispersed "hallmark" cells that contained eccentric kidney-shaped or embryo-like nuclei, several prominent rod shaped or angulated basophilic nucleoli, and abundant amphophilic cytoplasm. "Doughnut" cells, tumor cells with multilobated nuclei, and multinucleated giant cells with a wreath-like arrangement of nuclei occasionally were found. A small number of "plasmacytoid" tumor cells, nondescript small round tumor cells, and reactive polymorphs also was present. In contrast, "plasmacytoid" cells and nondescript small to medium-sized tumor cells represented the predominant cell population in ALCL of the small cell variant. The "plasmacytoid" appearance was exaggerated further in air-dried smears. In air-dried smears, small intracytoplasmic vacuoles and scanty azurophilic granules also were noted. On immunocytochemical study performed using the cell block materials, the majority of tumor cells demonstrated membranous and paranuclear "dot-like" positivity for CD30. The staining for epithelial membrane antigen, leukocyte common antigen, and T-cell markers was variable. Positive staining for ALK protein was demonstrated beautifully in two of the cases. CONCLUSIONS: Despite the wide morphologic spectrum of ALCL, a definitive diagnosis on the basis of FNA cytology is possible on careful interpretation of the cytologic features and a high index of suspicion. The cytologic diagnosis can be confirmed further with proper application of immunostaining to cell block sections. Immunocytochemical study for ALK protein, which provides useful prognostic information, also can be demonstrated satisfactorily using cytology samples. PMID- 12589645 TI - Cytologic criteria for well differentiated adenocarcinoma of the pancreas in fine needle aspiration biopsy specimens. AB - BACKGROUND: Fine-needle aspiration biopsy (FNAB) of pancreatic lesions has become popular to establish a tissue diagnosis before chemotherapy and/or surgery. The diagnostic sensitivity and specificity of FNAB have improved as a result of several articles regarding cytologic criteria for pancreatic adenocarcinoma. However, false-negative and "suspicious for malignancy" rates remain relatively high, in part because of the underdiagnosis of well differentiated adenocarcinoma (WDA). Existing cytologic criteria do not specifically focus on WDA. In this study, the authors attempt to add to, redefine, and test cytologic criteria for WDA of the pancreas in FNAB specimens. METHODS: The authors retrospectively reviewed the specimens of 291 consecutive computed tomography-guided FNABs of pancreatic lesions performed at the study institution between 1995 and 1999. The original cytologic diagnoses were confirmed by cell blocks (131), surgical tissue (84), or clinical follow-up. The FNAB specimens were evaluated for the presence of 10 cytologic criteria: 1) anisonucleosis, 2) nuclear membrane irregularity, 3) nuclear crowding/overlapping/three-dimensionality, 4) nuclear enlargement (if there are more than two red blood cells), 5) gap versus confluent cell spacing, 6) hyperchromasia, 7) macronucleoli, 8) mitosis, 9) chromatin clearing, and 10) necrosis. RESULTS: The original cytologic diagnoses were nondiagnostic in 24 cases, benign in 27, suspicious for malignancy in 15, and malignant in 225. Among the 225 malignant lesions, 74 cases were diagnosed as WDA. Cytologic criteria 1-4 were observed in 92-99% of WDA cases, whereas criteria 5-10 were present in only 7-38% of WDA cases. Six of 15 suspicious and 4 of 27 negative cases were of low cellularity but retrospectively met the cytologic criteria 1-4 for WDA (the diagnosis was confirmed on clinical follow-up). CONCLUSIONS: The diagnosis of pancreatic WDA can be made in FNAB specimens by the observance of anisonucleosis, nuclear membrane irregularity, nuclear crowding/overlapping/three-dimensionality, and nuclear enlargement. Necrosis, chromatin clearing, mitosis, macronucleoli, and hyperchromasia are of limited diagnostic significance because they are commonly absent in pancreatic WDA. PMID- 12589646 TI - The use of CDKN2A deletion as a diagnostic marker for malignant mesothelioma in body cavity effusions. AB - BACKGROUND: The distinction between benign reactive mesothelial cells and malignant mesothelial cells in serous effusions is difficult and has an unusually high false negative rate. Unfortunately, there are no generally accepted markers to distinguish between benign reactive and malignant mesothelial cells. Homozygous deletion of CDKN2A is frequent in mesothelioma (present in > 70% of tumors). Therefore, detection of CDKN2A deletion by fluorescence in situ hybridization (FISH) was evaluated as an ancillary test in the cytologic diagnosis of malignant mesothelioma. METHODS: Dual-color FISH for CDKN2A and chromosome 9 centromere was performed on cytolyt-fixed Thinprep slides from 6 cytologically suspicious and 7 cytologically positive effusions (all with histologically confirmed mesothelioma) and in 19 cytologically benign effusions (14 pleural effusions, 3 pericardial effusions, and 2 abdominal fluid specimens). Specimens containing > or = 15 nuclei that lacked signals for CDKN2A but showed at least 1 signal for chromosome 9 centromere were considered positive. In samples with negative cytology, the nuclei of at least 100 mesothelial cells were evaluated; whereas, in specimens with positive or suspicious cytology, counting nuclei was done only if < 15% of nuclei showed homozygous loss of CDKN2A. RESULTS: Homozygous deletion was detected in mesothelial cells in six of seven specimens with positive cytology and in six of six specimens with suspicious cytology. Cytologically, there were numerous tumor cells in a single positive specimen without homozygous deletion. All 19 cytologically negative specimens were negative for CDKN2A deletion. CONCLUSIONS: The detection of homozygous CDKN2A deletion by FISH would have been helpful in confirming a diagnosis of mesothelioma over reactive mesothelial cells in 12 of 13 samples with positive or suspicious cytology. Further studies on larger series of patients with suspicious cytology are needed to evaluate the validity and efficiency of this approach for improving the diagnostic accuracy of effusion cytology. PMID- 12589647 TI - Human papillomavirus typing and DNA ploidy determination of squamous intraepithelial lesions in liquid-based cytologic samples. AB - BACKGROUND: Infection by high-risk human papillomavirus (HPV) plays a role in the evolution of cervical carcinoma. Cellular atypia and consecutive DNA content alterations in cytologic samples are consequences of a preexisting viral infection. METHODS: We analyzed the frequency and association of HPV types and the presence of rare cells with abnormally high DNA content. We also evaluated whether these findings support the cytologic diagnosis in 112 routine cases with low and high-grade squamous intraepithelial lesions (LSIL/HSIL) when performed from liquid-based cytologic samples (ThinPrep). For DNA content measurements, laser scanning cytometry was applied and at least 10,000 cells were analyzed. HPV typing was performed by a direct sequencing approach using the consensus primers GP5+/GP6+ and MY09/MY11. RESULTS: Of 112 SIL cases, 110 (98.2%) were HPV positive and 95 (84.8%) had a high-risk type HPV infection. Almost one-half of the cases (46 of 95, 48.4%) with a high-risk HPV infection presented aneuploid squamous cells with greater than 9c DNA content, whereas none of the low-risk HPV-positive or HPV-negative SIL cases showed any aneuploid cells in this range. Although 91.8% of the HSIL cases displayed greater than 9c aneuploid cells, only 7.9% of the LSIL cases were positive for such cells with abnormally high DNA content. CONCLUSIONS: HPV typing and DNA measurements help in the objectivation of cytologic atypia and both can be performed efficiently from the same liquid-based cytologic samples. PMID- 12589648 TI - Surface, core, and X genes of hepatitis B virus in hepatocellular carcinoma: an in situ hybridization study. AB - BACKGROUND: The incidence of hepatocellular carcinoma (HCC) and the seroprevalence of hepatitis B virus (HBV) in this disease state are significantly higher in South India than in North India. Because data on serologic studies do not project the actual association between the two parameters, this study was undertaken. METHODS: The prevalence of HBV genes in HCC patients was studied using nonisotopic in situ hybridization. Fifty patients from South India were diagnosed with HCC after performing ultrasound-guided fine-needle aspiration biopsies of liver lesions. The diagnosis was confirmed by cell block studies. Sections cut from paraffin-embedded cell blocks made out of the aspirates were probed with digoxigenin-labeled surface, core, and X regions of the viral genome. RESULTS: Nuclear integration of the surface gene was observed in 100% (50 of 50), the core gene was positive in 94% (47 of 50), and the X gene was present in 98% (49 of 50) of the cases. An episomal form of the virus was not found. Serum hepatitis B surface antigen was positive only in 48% (12 of 25) of the patients screened. CONCLUSIONS: We found molecular evidence that HBV is an important contributing factor in the etiology of HCC in South India. In HCC, the S gene of the virus was the most prevalent followed by the X and C genes. Only integrated forms of the viral DNA were observed. Nonisotopic in situ hybridization using multiple regions of the viral genome is a good technique for studying this association. It has an added advantage over polymerase chain reaction, of localization of signals in a tumor cell. Cell blocks made from fine-needle aspirates are ideal for in situ hybridization. PMID- 12589649 TI - c-fos and estrogen receptor gene expression pattern in the rat uterine epithelium during the estrous cycle. AB - Different studies in ovariectomized estrogen treated animals support the idea that c-fos plays a role in the proliferation of uterine epithelial cells. However, these studies invite us to reassess the role played by c-fos in epithelial cell types of the endometrium during the estrous cycle. The present study was undertaken to determine the c-fos and estrogen receptor (ER) gene expression pattern in the rat uterine epithelium during the estrous cycle in which natural and cyclic changes of steroid hormones occur, and correlate these changes with the proliferation status of this cellular types. Proliferation was assessed during the estrous cycle using bromodeoxyuridine incorporation to DNA. ERalpha and beta proteins were assessed by immunohistochemistry. The regulation of c-fos gene expression in the uterus of intact animals during the estrous cycle was evaluated using both in situ hybridization and immunohistochemistry. Estradiol (E(2)) and progesterone (P(4)) plasma levels were assessed by radioimmunoassay. The results indicated that luminal (LE) and glandular epithelia (GE) presented maximal proliferation during the metestrus (M) and the diestrus (D) days. However, during the proestrus (P) day only LE presented proliferation, and during the estrus (E) day only the stromal cells proliferated. A marked immunostaining for ERalpha was detected in both LE and GE cells during the early phases of the cycle but diminished on the P and the E day. In contrast, ERbeta was undetectable in both epithelia during all stages of the cycle. The highest c fos mRNA level was detected in both epithelia on the M day, followed by a significant reduction during the other days of the cycle. The highest protein content was observed on the M and D days, and the minimal value was detected on the E day. The c-Fos protein level in LE was increased during M and D days, presenting a high correlation with the cellular proliferation pattern of this cell type. In conclusion, the overall results indicate that c-Fos protein presented a good correlation with uterine epithelial cell proliferation of LE. In the case of GE, the same tendency was observed, although no significant correlation was found. Both in LE and GE, c-fos mRNA did not strictly correlate with its protein levels. c-fos seems to have a postranscriptional regulation in uterine epithelial cells during the rat's estrous cycle. PMID- 12589650 TI - Stage-specific regulatory element of mouse Sry gene. AB - Sry expression is essential for initiating male sex differentiation, and the expression occurs only during a restricted period in the developing gonad. It is thought that Sry is part of a pathway of genes that regulate sex determination. Although the interactions of several genes with Sry expression have been suggested, the exact cascade of gene expression regulating Sry transcription is entirely obscure because there is no available cell line expressing Sry and reflecting an in vivo condition. The present study was carried out to investigate the cis-acting element of the mouse Sry that responds stage specifically to its expression, in part, using transgenic mice expressing GFP on the Y chromosome. Ten DNA fragments were generated by digesting the 5' upstream region (positions 5491-8039; 2,549 bp) of mouse Sry with appropriate restriction enzymes. In an electrophoretic mobility assay with these fragments, the region from position 5491 to position 5799 (309 bp) was identified as forming specific protein-DNA complexes with nuclear extracts from 11.5 days post coitus (dpc) gonads, but not from 12.5 and 13.5-dpc gonads. This region also formed specific protein-DNA complexes with the nuclear extracts from adult testicular germ cells that generate only a circular form from Sry. This stage-specific responsive region was narrowed down to positions 5559-5616 by DNase I footprinting analysis. The assay of DNase I hypersensitive (HS) using the nuclear lysates from the 11.5-dpc urogenital ridges demonstrated that the novel HS site was located in the proximity of position 5600. This region DNase I HS was also detected at the same position when the lysates from adult testicular germ cells were applied. The results indicate that the present HS site may be involved in the transcriptional regulation of the linear and/or circular molecule transcripts from mouse Sry gene. PMID- 12589651 TI - Over-expression and secretion of angiogenin in intrauterine growth retardation placenta. AB - Human angiogenin is a potent inducer of neovascularization. There is a strong evidence to suggest that it might be involved in morphological and angiogenic changes in the placenta, that are necessary for a successful fetal outcome during pregnancy. However, its precise role in the pathogenesis of abnormal pregnancies is yet unknown. Intrauterine growth retardation (IUGR), an abnormal pregnancy is not a specific disease entity per se, but rather a manifestation of many possible fetal and maternal disorders. In this study, we demonstrated, for the first time, that placental explants in vitro secrete significantly elevated levels of angiogenin in placental tissues from patients with IUGR. We also observed enhanced mRNA expression in placenta from these patients. In addition, using the immunohistochemical methods, we observed identical staining of angiogenin to villous syncytiotrophobalst and fetal endothelial cells in both IUGR and normal placenta. Functionally active placental explants were used to detect immunoreactive angiogenin in conditioned media of all the samples from IUGR placenta and normal term group. The mean levels of angiogenin secreted by IUGR placenta were 1.4-, 1.6-, and 1.3-fold higher (P < 0.01) than normal term samples at 24, 48, and 72 hr of culture, respectively. Expression profiles of angiogenin from term and IUGR cases are in agreement with its mRNA levels and immunoblot analysis. In conclusion, the significant elevated levels of angiogenin in IUGR placenta may provide a molecular mechanism for the abnormal placental development. PMID- 12589652 TI - Identification of estrogen-regulated genes in the mouse uterus using a delayed implantation model. AB - Gonadal steroid hormones are known to modulate the implantation of the blastocyst, but how the controlling genetics are regulated remains largely unknown. Using a delayed-implantation model, we examined estrogen-regulated genes (ERGs) in the mouse uterus using the differential-display reverse transcription polymerase chain reaction (DD RT-PCR). Pregnant mice were ovariectomized and injected daily with progesterone (P, 1 mg/mouse), followed by a single injection of estrogen (E, 200 ng/mouse); 24 or 48 hr later, total RNA was extracted from the uterus. Reverse Northern analysis verified the expression patterns of 36 clones out of thousands of RNA species. Only five clones had mRNA levels that were modified, whereas other mRNAs were unchanged or not detectable. Sequence analysis of these, using the Basic Local Alignment Search Tool (BLAST) service, revealed that four of these clones were novel; one clone, designated ERG10, was found to be the mouse homologue of that deleted in oral cancer DOC-1. DOC-1 mRNA was detected all tissues examined, but only in the uterus and cervix was markedly increased 12 hr after E administration, it returned to basal level by 48 hr. One of the novel genes, designated ERG8, had three different forms of mRNAs and was expressed ubiquitously in all examined tissues. In the uterus, the mRNA level of ERG8 also increased 12 hr after E administration. These results suggest that during the implantation process, E differentially regulates several genes depending on cell type. Uterine-specific induction of newly found genes, such as ERG8 and 10, by E appears to be important for the early implantation process. PMID- 12589653 TI - ADAM7 (a disintegrin and metalloprotease 7) mRNA is suppressed in mouse epididymis by neonatal exposure to Diethylstilbestrol. AB - We investigated whether neonatal exposure to diethylstilbestrol (DES) induces the alteration of mRNA expression in adult mouse epididymis, which plays an important role in sperm maturation. Using a cDNA subtraction method, we isolated 15 changed gene clones in neonatally DES-treated mouse epididymides, and we found a clone homologous with a disintegrin and metalloprotease (ADAM) 7 in the epididymis, as a suppressed gene, by means of neonatal DES treatment in 8-week-old mice. Indeed, it was confirmed by Northern blot analysis that the ADAM7 mRNA expression in the epididymis was at a lower level in neonatally DES-treated mice than in non treated mice. Moreover, in situ hybridization analysis and real-time reverse transcription and polymerase chain reaction (real-time RT-PCR) revealed that ADAM7 expression was markedly reduced in the corpus region of the epididymis of DES-treated mice as compared with non-treated mice. Our results suggest that neonatal exposure to DES leads to the suppression of ADAM7 expression in the epididymis in the long term. ADAM7 gene expression might be a biological marker of fetal or neonatal exposure to estrogenic compounds, including endocrine disruptors. PMID- 12589654 TI - Germ cell transplantation in goats. AB - Transplantation of spermatogonial stem cells provides a unique approach for the study of spermatogenesis and manipulation of the male germ line. This technique may also offer an alternative to the currently inefficient methods of producing transgenic domestic animals. We have recently established the technique of spermatogonial transplantation, originally developed in laboratory rodents, in pigs, and this study was aimed to extend the technique to the goat. Isolated donor testis cells were infused into the seminiferous tubules of anesthetized recipient goats through an ultrasonographically-guided catheter inserted into the rete testis. Donor cells were obtained by enzymatic digestion of freshly collected testes from immature goats (either from the recipients' contralateral testis or from unrelated donors). Prior to transplantation, testis cells were labeled with a fluorescent marker to allow identification after transplantation. Recipient testes were examined for the presence and localization of labeled donor cells at 3-week intervals up to 12 weeks after transplantation. Labeled donor cells were found in the seminiferous tubules of all testes, comprising 10-35% of the examined tubules. Histological examination of the recipient testes did not reveal evident tissue damage, except for limited fibrotic changes at the site of needle insertion. Likewise there were no detectable local or systemic signs of immunologic reactions to the transplantations. These results indicate that germ cell transplantation is technically feasible in immature male goats and that donor-derived cells are retained in the recipient testis for at least three months and through puberty. This study represents the first report of germ cell transplantation in goats. PMID- 12589655 TI - Proliferation of donor mitochondrial DNA in nuclear transfer calves (Bos taurus) derived from cumulus cells. AB - In embryos derived by nuclear-transfer (NT), fusion of donor cell and recipient oocyte caused mitochondrial heteroplasmy. Previous studies from other laboratories have reported either elimination or maintenance of donor-derived mitochondrial DNA (mtDNA) from somatic cells in cloned animals. Here we examined the distribution of donor mtDNA in NT embryos and calves derived from somatic cells. Donor mitochondria were clearly observed by fluorescence labeling in the cytoplasm of NT embryos immediately after fusion; however, fluorescence diminished to undetectable levels at 24 hr after nuclear transfer. By PCR mediated single-strand conformation polymorphism (PCR-SSCP) analysis, donor mtDNAs were not detected in the NT embryos immediately after fusion (less than 3 4%). In contrast, three of nine NT calves exhibited heteroplasmy with donor cell mtDNA populations ranging from 6 to 40%. These results provide the first evidence of a significant replicative advantage of donor mtDNAs to recipient mtDNAs during the course of embryogenesis in NT calves from somatic cells. PMID- 12589656 TI - Maternal gamma (gamma)-tubulin is involved in microtubule reorganization during bovine fertilization and parthenogenesis. AB - In this study, gamma-tubulin distribution was determined chronologically in conjunction with microtubule dynamics during bovine fertilization and parthenogenesis. In unfertilized bovine oocytes, gamma-tubulin was identified in the cytoplasm, mainly in the cortex and concentrated in the meiotic spindle. Following sperm penetration, gamma-tubulin in the cytoplasm was recruited by a sperm component. During pronuclear apposition, gamma-tubulin was localized as spots at the spindle poles. gamma-tubulin spots were observed in blastomeres of embryos cleaved in vitro. Following electrical stimulation, gamma-tubulin and microtubule matrix were noted in oocyte cortex. In the late pronuclear stage, considerably less gamma-tubulin and microtubules were detected in the cytoplasm. At the mitotic metaphase of parthenotes, gamma-tubulin was recruited to the condensed chromatin and concentrated in the spindle. The gamma-tubulin spots were not detected until the 8-cell stage of parthenotes. This suggests that maternal gamma-tubulin is recruited by a sperm component to reconstitute the zygotic centrosome. In the absence of sperm components, the cell cycle-related assembly of gamma-tubulin organizes microtubule nucleation for positioning the pronucleus and spindle protein of mitotic metaphase during the first cell cycle of bovine parthenotes. PMID- 12589657 TI - Development of the signalling pathways associated with sperm capacitation during epididymal maturation. AB - As spermatozoa mature within the epididymis they acquire the potential for capacitation and ultimately fertilization. In biochemical terms, the former is reflected in the progressive activation of a signal transduction pathway characterized by cAMP-mediated induction of phosphotyrosine expression on the sperm tail. In this study, we have examined the cellular mechanisms controlling this maturational event. Caput epididymal spermatozoa exhibited tyrosine phosphorylation on the sperm head that was largely unresponsive to cAMP and not significantly impaired by removal of extracellular HCO(3) (-). In contrast, caudal epididymal spermatozoa exhibited low levels of phosphorylation on the sperm head, yet responded dramatically to cAMP by phosphorylating a new set of proteins on the sperm tail via mechanisms that were highly dependent on extracellular HCO(3) (-). The impact of extracellular HCO(3) (-) depletion on caudal cells was not associated with a significant change in the redox regulation of cAMP but could be fully reversed by buffering the intracellular pH with N Tris[Hydroxymethyl]methyl-3-amino-propanesulfonic acid (TAPS). The pattern of tyrosine phosphorylation was also profoundly influenced by the presence or absence of added extracellular calcium. In the presence of this cation, only caudal spermatozoa could respond to increased extracellular cAMP with tyrosine phosphorylation of the sperm tail. However, in calcium-depleted medium, this difference completely disappeared. Under these conditions, caput and caudal spermatozoa were equally competent to exhibit phosphotyrosine expression on the sperm tail in response to cAMP. These results emphasize the pivotal role played by calcium and HCO(3) (-) in modulating the changes in tyrosine phosphorylation observed during epididymal maturation. PMID- 12589658 TI - Chromatin configuration and transcriptional control in human and mouse oocytes. AB - In vitro maturation of human oocytes at the germinal vesicle (GV) stage could offer an alternative in several cases of female infertility. It however rests on a better knowledge of the quality of human oocyte. Using fluorescence imaging of DNA and of the transcription sites, combined with electron microscopy, we show that human oocytes follow size-dependent changes in chromatin configuration, transcription sites distribution and nuclear ultrastructure that follow those observed in mouse GV oocytes. We thus analyzed in mouse GV oocytes the phosphorylation dependence of the transcriptional activity. We show by Western blot that, while active GV oocytes have approximately the same proportion of hypo and hyperphosphorylated forms of the RNA polymerase II (RNAP II), the hyperphosphorylated form is almost absent from inactive oocytes. We also show that (1) RNAP II-dependent transcription is much less sensitive to various kinase inhibitors in mouse oocytes than in somatic cells or mouse one-cell embryos, although the phosphorylation equilibrium of RNAP II was largely shifted towards the hypo-phosphorylated form upon treatment with these inhibitors (2) RNAP I is completely insensitive to kinase inhibitors in GV oocytes. PMID- 12589659 TI - Meiosis in male PL/J mice: a genetic model for gametic aneuploidy. AB - Sperm from mice of the PL/J strain have a high frequency of sperm-head morphology abnormalities. Fluorescence in situ hybridization (FISH) methods revealed that PL/J sperm are also characterized by a high frequency of aneuploidy. The traits of abnormal sperm head morphology and aneuploidy are associated with numerous meiotic abnormalities. Spermatocytes of PL/J mice exhibit chromosome asynapsis during meiotic prophase as well as reduced crossing over, revealed by analysis of both MLH1 foci in pachytene spermatocytes and chiasmata seen at the first meiotic metaphase. During the first meiotic division, roughly one-third of the PL/J spermatocytes exhibit aberrant spindle morphology, with abnormalities including monopolar spindles, split spindle poles, and incomplete spindle formation and centrosomal abnormalities. F1 progeny of a cross between PL/J and C57BL/6J did not exhibit a high frequency of either sperm aneuploidy or sperm head morphology aberrations, as would be expected if the PL/J traits were dominant. Among progeny of a backcross of F1 mice to PL/J, none of 16 males assessed exhibited elevated frequencies of sperm head morphology abnormalities. Four of the individuals exhibited elevated sperm aneuploidy, but not at the levels of the PL/J parents. Thus, it is likely that the aberrant PL/J traits are due to several genes and/or modifiers affecting the generation of both sperm aneuploidy and abnormal sperm head morphology. PMID- 12589660 TI - Alterations and reversibility in the chromatin, cytoskeleton and development of pig oocytes treated with roscovitine. AB - Germinal vesicle (GV) breakdown in mammalian oocytes is regulated by the activation of maturation promoting factor (MPF). We investigated a specific cdc2 kinase inhibitor, roscovitine, to maintain pig oocytes in the GV stage. Cumulus oocyte complexes (COCs) were aspirated from slaughterhouse ovaries and cultured for 44 hr in NCSU#23 medium containing different levels of roscovitine (0, 10, 20, 30, 40, 50 microM in Experiment 1 and 0, 40, 60, 80, 100, 120 microM in Experiment 2). The COCs were cultured for another 44 hr after removal of the chemical. Twenty oocytes in each group were fixed at 44 hr for immunocytochemical labeling of the cytoskeleton and the rest (approximately 20/group) were fixed at the end of 88 hr after culture. Results showed that the inhibition of the oocyte in the GV stage was not effective when 10-50 microM (Experiment 1) of roscovitine were used (19-34%). When oocytes were released from the inhibitor, similar proportions (70-83%) of oocytes were observed in the MII or advanced stages among treatments. However, when higher concentrations of roscovitine were used (Experiment 2), significantly greater inhibitory effect was observed at the levels of 80-120 microM with 83-91% oocytes being blocked in the GV stage when compared to the control (9%) and the 40-60 microM (27-43%) groups (P < 0.05). Although 15-21% of the oocytes showed abnormal MII morphology with aberrant meiotic spindles and/or formation of cytoplasmic microtubules, a substantial number of oocytes resumed meiosis and reached MII stage at 44 hr after removal of this chemical. In Experiment 3, different concentrations of roscovitine (0, 20, 40, and 80 microM) were tested to examine the length of intervals (0, 11, 22, 33, and 44 hr) for an effective inhibition. Results showed that the inhibitory effect was significantly more prominent at 22 hr than that at 33 and 44 hr after roscovitine treatment in all treatment groups (P < 0.05). This study demonstrated that roscovitine-treated oocytes resumed meiosis after removal of the inhibitor. This could provide flexibility for studying porcine oocyte development and embryo cloning and may have application in other species. PMID- 12589661 TI - Glutathione and adenosine triphosphate content of in vivo and in vitro matured porcine oocytes. AB - Glutathione (GSH) content in mature porcine oocytes is correlated with subsequent fertilization and developmental success. Adenosine triphosphate (ATP) is an important energy source for maintaining cellular activities and protein synthesis. The objective of this study was to compare GSH and ATP concentrations of in vivo and in vitro matured porcine oocytes. Ovulated, in vivo matured oocytes were frozen at -80 degrees C in groups of 10-20 (GSH) or 5-10 (ATP). In vitro oocytes were matured in either tissue culture medium-199 (TCM199) supplemented with polyvinyl alcohol (PVA) or hyaluronic acid (MAP5), or North Carolina State University-23 (NCSU23) supplemented with porcine follicular fluid (pFF) and frozen as described, or fertilized and cultured. GSH content was determined by the dithionitrobenzoic acid-glutathione disulfide (DTNB-GSSG) reductase recycling assay. ATP content was determined by using the Bioluminescent Somatic Cell Assay Kit. Oocytes matured in vitro in defined TCM199 with PVA or hyaluronic acid, or NCSU23 with pFF had significantly lower concentrations (P < 0.05) of GSH (n = 207, 9.82 +/- 0.71 pmol/oocyte; n = 104, 9.73 +/- 0.81 pmol/oocyte; n = 108, 7.89 +/- 0.66 pmol/oocyte, respectively) compared to in vivo matured oocytes (n = 217, 36.26 +/- 11.00 pmol/oocyte). Concentrations of ATP were not different between treatments (in vivo, n = 70, 0.97 +/- 0.07 pmol/oocyte; TCM-PVA, n = 117, 0.81 +/- 0.13 pmol/oocyte; TCM-MAP, n = 107, 1.02 +/- 0.18 pmol/oocyte; NCSU-pFF, n = 134, 0.71 +/- 0.08 pmol/oocyte). Intracellular ATP content does not appear to be related to developmental potential in porcine oocytes. Low intracellular GSH may be responsible, in part, for lower developmental competence observed in in vitro matured porcine oocytes. PMID- 12589662 TI - Increased recombination frequency showing evidence of loss of interference is associated with abnormal testicular histopathology. AB - Nondisjunction leading to aneuploid gametes has been linked genetically to both increases and decreases in recombination frequency on the aneuploid chromosome. In the present study, we present physical evidence of increased frequency of recombination nodules as measured by Mut-S-like homologue-1 (MLH1) foci on pachytene chromosomes from sterile male mice homozygous for a mutation in the protein phosphatase 1cgamma (PP1cgamma) gene. The pattern of elevated recombination frequency in PP1cgamma mutant spermatocytes is consistent with a loss of interference. Previous studies demonstrated: (1) spermiogenesis is impaired starting at step 8 with a severe reduction in elongating and condensed spermatids; (2) spermatids and sperm exhibit elevated rates of DNA fragmentation; and (3) haploid gametes exhibit elevated levels of aneuploidy. Morphometric analysis of developing testes revealed that the first wave of meiosis proceeds at a normal rate in mutant testes, a surprising result given that the PP1 inhibitor okadaic acid has been shown to accelerate progression of spermatocytes from pachytene to the first meiotic division (MI). Evidence of abnormal testicular histopathology is apparent at 3 weeks, before the appearance of haploid gametes, eliminating the possibility that the mutant phenotype is caused by the presence of abnormal spermatids, but coincident with the appearance of the first set of mid to late pachytene spermatocytes. These observations lead us to conclude that the PP1cgamma mutation causes a complex phenotype, including subtle adverse effects on meiosis, possibly mediated by defective signaling between germ cells and Sertoli cells. PMID- 12589663 TI - Galen on the anatomy and treatment of the injured hand. PMID- 12589664 TI - Variations in the number and position of human lumbar sympathetic ganglia and rami communicantes. AB - The purpose of this study is to provide precise anatomical and statistical information about the number and location of lumbar sympathetic ganglia and the number and length of the related rami communicantes, and to consider the neurological pathway for nociception from the low back. Three-hundred ninety three ganglia and 719 rami communicantes from 50 human cadavers were identified by gross dissection. The number of ganglia in a single lumbar chain ranged from 2 to 6, the mean being 3.9. The mean lengths of rami connected to the 1st and 5th lumbar spinal nerves, respectively, were significantly longer and shorter than those connected to the 2nd, 3rd, and 4th nerves. The lumbar sympathetic ganglia and rami communicantes were not distributed segmentally. The present results may assist in understanding the nociceptive pathway from the low back. PMID- 12589665 TI - What arteries are detectable in the precarinal space on contrast-enhanced CT? AB - Although the precarinal space is critical in the interpretation of computed tomography (CT) of the thorax, vascular structures within the space have not been well documented. The aim of this study was to investigate vascular structures in the precarinal space that are detectable with contrast-enhanced CT scans. Contrast-enhanced CT chest scans from 90 patients aged 17-78 years (41 male, 49 female) were analyzed retrospectively. Twenty-two cadavers aged 54-93 years (13 male, 9 female) were used for gross anatomic study. We found that 9 of 90 cases of the contrast-enhanced CT scans had vessel-like structures detectable in the precarinal space, located primarily anterior to the right main bronchus. Single or double arteries were identified in the precarinal space of 19 cadavers. Thirteen of the 23 arteries observed were the right bronchial artery and 10 were the left bronchial artery. In two cadavers, a single right bronchial artery (2.5 and 4.0 mm in diameter), which could have been detected with contrast-enhanced CT, originated from the aortic arch and the root of the left subclavian artery and coursed in the precarinal space. Based on the variation in the location and size of the bronchial arteries observed in this study, we suggest that vascular structures are detectable in the precarinal space with contrast-enhanced CT, particularly anterior to the right main bronchus. PMID- 12589666 TI - Topography of the pelvic autonomic nervous system and its potential impact on surgical intervention in the pelvis. AB - Bladder, bowel, and sexual dysfunction caused by iatrogenic lesions of the inferior hypogastric plexus (IHP) are well known and commonly tolerated in pelvic surgery. Because the pelvic autonomic nerves are difficult to define and dissect in surgery, and their importance often ignored, we conducted a gross anatomic study of 90 adult and four fetal hemipelves. Using various non-surgical approaches, the anatomic relations and pathways of the IHP were dissected. The IHP extended from the sacrum to the genital organs at the level of the lower sacral vertebrae. It originated from three different sources: the hypogastric nerve, the sacral splanchnic nerves from the sacral sympathetic trunk (mostly the S2 ganglion), and the pelvic splanchnic nerves, which branched primarily from the third and fourth sacral ventral rami. These fibers converge to form a uniform nerve plate medial to the vascular layer and deep to the peritoneum. The posterior portion of the IHP supplied the rectum and the anterior portion of the urogenital organs; nerve fibers traveled directly from the IHP to the anterolateral wall of the rectum and to the inferolateral and posterolateral aspects of the urogenital organs. The autonomic supply from the IHP was supplemented by nerves accompanying the ureter and the arteries. An understanding of the location of the autonomic pelvic network, including important landmarks, should help prevent iatrogenic injury through the adoption of surgical techniques that reduce or prevent postoperative autonomic dysfunction. PMID- 12589667 TI - Anatomy of the arcus tendineus fasciae pelvis in females. AB - Because of its proximity to the urethra, the anterior part of the arcus tendineus fasciae pelvis (ATFP) may be used in urethrosuspension procedures for urinary stress incontinence. In this study, 10 embalmed female cadaver hemipelves were dissected and their gross anatomy described. In females, the ATFP is a condensation of the endopelvic fascia. The anterior attachment of the ATFP is to the caudal inner surface of the body of the pubic bone at a site averaging 4 mm lateral to the pubic symphysis and covering an average area of 53 mm(2). Posteriorly, it attaches to the medial surface of the ischial spine. In nine of the 10 hemipelves the first anterior centimeters of the ATFP have a clear lateral fixation either to the lateral part of the levator ani muscle (n = 1), to the fascia covering the obturator internus muscle (n = 7), or to the obturator membrane (n = 1). Medially from the ATFP derives a 2-3-cm long flat fibrous attachment to the posterolateral aspect of the urethra. In eight unembalmed cadavers, the ATFP gave way at a pulling force of 8.2 kg (range = 3.5-11.5 kg). The ATFP resists caudal movement of the proximal anterior vaginal wall and the urethra in the upright posture and, therefore, may be suitable for urethrosuspension procedures. PMID- 12589668 TI - Identification of a sphincter at the sigmoidorectal canal in humans: histomorphologic and morphometric studies. AB - In a recent study we demonstrated that the rectosigmoid junction is a large bowel segment 2.8 +/- 0.9 cm in length, which we have termed the 'sigmoidorectal canal' (SRC). In that study we showed that the SRC exerted increased intraluminal pressure, contained a thick, circular, smooth muscle layer, and reacted to sigmoid colon (SC) and rectal contraction by relaxation and contraction, respectively. These findings suggest the existence of an anatomical sphincter at the SRC. To investigate this hypothesis, in the present study we carried out a morphometric (quantitative) analysis of the musculature of the SC, SRC, and rectum. Twenty-two cadavers (13 male, 9 female; mean age = 32.3 +/- 17.6 years) were investigated. Histomorphologic studies comprised staining transverse (15 cadavers) and longitudinal (7 cadavers) sections from the SC, SRC, and rectum. Specimens were stained with H&E, Masson's trichrome, and silver stains. Morphometric studies, using image analyzer computer equipment, measured the muscle thickness of the muscularis externa (ME) of the SC, SRC, and rectum. The ME of the SRC was markedly thickened compared to the ME of the SC or rectum. In 16 specimens, the increased thickness involved mainly the circular muscle layer and in six specimens both the circular and longitudinal layers. Connective tissue septa connected the adventitia with the submucosa: these 'adventitial septa' contained nerve ganglia. 'Localized bundles' of smooth muscle fibers were detected between the circular muscle fibers. The morphometric findings showed that the mean thickness of the circular muscle layer measured 440.3 +/- 80.1 microm in the SC, 1166.1 +/- 255.4 microm in the SRC, and 394.9 +/- 123.9 microm in the rectum. This study strongly suggests the presence of an anatomic sphincter at the SRC. This sphincter may explain the higher pressure at the SRC and the response of the SRC to rectal and SC contractions. The localized smooth muscle bundles in the ME may represent a rectosigmoid pacemaker. PMID- 12589669 TI - Lumbar lordosis: study of patients with and without low back pain. AB - We used magnetic resonance imaging (MRI) to assess lumbar lordosis in 27 patients with low back pain and 19 patients and 10 volunteers with no known back pain. Our study aimed to investigate whether lordosis changes with age and is reduced in those with low back pain. Although our results confirm known observations that lumbar lordosis is more prominent in women (P < 0.01) and those with a higher body mass index (P < 0.04), we were unable to demonstrate any significant variation in lordosis with age. Nor could we demonstrate any difference in the degree of lordosis among women with or without back pain. Men with low back pain tended to have a less prominent lordosis, but this difference did not reach statistical significance. Therefore, a 'reduced lumbar lordosis' should be regarded as a very weak clinical sign. PMID- 12589670 TI - Incidence of metopism in the Lebanese population. AB - This study was carried out on 968 skull X-rays to determine the incidence of the metopic suture in the Lebanese population. Complete and incomplete metopism was present in 0.82% and 0.93% of cases, respectively, leading to an overall incidence of 1.75%. In both categories, the incidence was relatively higher in males (1.84%) than in females (1.62%). Persons living in rural areas had a higher incidence of complete and incomplete metopism compared to persons living in urban areas, with ratios of 4:1 and 4:2, respectively. Other findings included an absence of frontal sinuses in all but one skull with complete metopism, accessory ossicles in the majority of cases of complete metopism (7/8), and hydrocephalus in one case. Medical records did not reveal any other associated diseases or abnormalities. Despite the low incidence of metopic suture in the Lebanese population, in X-ray diagnosis of fractures of the frontal bone, metopic suture must not be ruled out. PMID- 12589671 TI - Investigation of connective tissue attachments to the cervical spinal dura mater. AB - The connective tissue attachments to the cervical spinal dura mater originating from the ligamentum nuchae (LN) and rectus capitis posterior minor (RCPM) muscle were evaluated in 30 cadaveric spines. Magnetic resonance images (MRIs) were correlated with the attachments in four cadaveric specimens. Attachments from the LN to the RCPM were also identified. The LN and the RCPM to dura attachments were found in all 30 specimens. Our results indicate that: 1) the attachments between the LN and RCPM and the dura occur between vertebrae C1-C2 and the occipital bone and C1, respectively, and that they are substantial normal anatomic attachments, 2) attachments between the LN and RCPM are usually present, and 3) the attachments between the LN and dura mater can be identified on MRI. These latter attachments may play a role in neck pain, making their MRI appearance clinically important. PMID- 12589672 TI - Seminar in breast self-examination for female medical students integrated into a human gross anatomy course. AB - A breast self-examination (BSE) seminar for first-year female medical students is presented and a single-gender approach for other subjects in the medical curriculum is discussed. In 1999 a small group seminar on BSE was offered at the Hannover Medical School to female medical students as part of their curriculum in human gross anatomy. An evaluation questionnaire was answered by 94 students (87% of participants). Frequencies of answers to two open questions were used as indicators of: 1) the acceptance of a single-gender course and 2) an increased awareness of breast cancer prevention. A linear regression analysis was carried out to identify the most important predictors for the global course evaluation and a heightened interest in breast cancer prevention. The mean global rating of the seminar was 13.8 (minimum: 1 point; maximum: 15 points). Factors that significantly influenced the global rating were the course atmosphere, the teacher's enthusiasm, and the professional interest of the students. An increased concern for breast cancer prevention was significantly dependent on the professional interest and the self-awareness of the women. The results suggest that there is a need for single-gender seminars in academic medicine and that instruction of female students in BSE is an ideal subject for this approach. Because of the prevalence of breast cancer, it is recommended that such a seminar become an integral part of the preclinical curriculum for all female medical students. PMID- 12589673 TI - Attitudes of medical and dental students to dissection. AB - Guy's, King's, and St. Thomas's School of Medicine encourages students to learn anatomy from human dissection. Today, there is a worldwide move of anatomy-based teaching away from dissection to prosection. This study investigates how attitudes toward dissection vary with gender and ethnicity. We assessed students' reactions and concerns regarding the dissecting room, any coping strategies they use to combat them, and analyzed effective methods of teaching anatomy to medical and dental students. Three questionnaires were distributed amongst 474 first-year medical and dental students before dissection and 1 week and 12 weeks after exposure to the dissecting room. Over the 3 months we found significant changes in the concerns of students about dissection. There were also significant differences (P < 0.05) between medical and dental students, males and females, and students of differing ethnic backgrounds, which persisted over 12 weeks. Both medical and dental students found tutorials and textbooks of most value in learning anatomy. Dental students found prosection more useful than medical students (P < 0.001) though neither group demonstrated a significant preference for prosection over dissection. Of concern, 7% reported recurring images of cadavers and 2% insomnia after commencing dissection. Interest in the subject matter and discussion were the commonest methods used to combat stress. This study contributes to the ongoing debate about the value of the dissecting room in the medical school curriculum. PMID- 12589674 TI - Unusual case of small bowel obstruction: persistence of vitelline artery remnant. PMID- 12589675 TI - Simultaneous occurrence of a thyromediastinal muscle, a truncus bicaroticobrachialis, and a left superior vena cava. AB - A case is presented of a combination of anatomical anomalies found in a 67-year old female cadaver during routine dissection by medical students. They include a thyromediastinal muscle, a truncus bicaroticobrachialis, and a left superior vena cava, with complete absence of the right superior vena cava, but with a normal azygos vein opening into the right atrium at the expected site of entry of the superior vena cava. No associated congenital cardiac malformations were found. Clinical implications include the difficulty of heart catheterization through the subclavian veins and misleading images on CT or MRI scans, where the azygos vein could be mistaken for a right superior vena cava. PMID- 12589677 TI - Sternomastoid paradox. PMID- 12589678 TI - Growth cones contain a dynamic population of neurofilament subunits. AB - Neurofilaments (NFs) are classically considered to transport in a primarily anterograde direction along axons, and to undergo bulk degradation within the synapse or growth cone (GC). We compared overall NF protein distribution with that of newly expressed NF subunits within NB2a/d1 cells by transfection with a construct encoding green fluorescent protein (GFP) conjugated NF-M subunits. GCs lacked phosphorylated NF epitopes, and steady-state levels of non phosphosphorylated NF subunits within GC were markedly reduced compared to those of neurite shaft as indicated by conventional immunofluorescence. However, GCs contained significant levels of GFP-tagged subunits in the form of punctate or short filamentous structures that in some cases exceeded that visualized along the shaft itself, suggesting that GCs contained a relatively higher concentration of newly synthesized subunits. GFP-tagged NF subunits within GCs co-localized with non-phosphorylated NF immunoreactivity. GFP-tagged subunits were observed within GC filopodia in which steady-state levels of NF subunits were too low to be detected by conventional immunofluorescence. Selective localization of fluorescein versus rhodamine fluorescene was observed within GCs following expression of NF-M conjugated to DsRed1-E5, which shifts from fluorescein to rhodamine fluorescence within hours after expression; axonal shafts contained a more even distribution of fluorescein and rhodamine fluorescence, further indicating that GCs contained relatively higher levels of the most-recently expressed subunits. GFP-tagged structures were rapidly extracted from GCs under conditions that preserved axonal structures. These short filamentous and punctate structures underwent rapid bi-directional movement within GCs. Movement of GFP tagged structures within GCs ceased following application of nocodazole, cytochalasin B, and the kinase inhibitor olomoucine, indicating that their motility was dependent upon microtubules and actin and, moreover, was due to active transport rather than simple diffusion. Treatment with the protease inhibitor calpeptin increased overall NF subunits, but increased those within the GC to a greater extent than those along the shaft, indicating that subunits in the GC undergo more rapid turnover than do those within the shaft. Some GCs contained coiled aggregates of GFP-tagged NFs that appeared to be contiguous with axonal NFs. NFs extended from these aggregates into the advancing GC as axonal neurites elongated. These data are consistent with the presence of a population of dynamic NF subunits within GCs that is apparently capable of participating in regional filament formation during axonal elongation, and support the notion that NF polymerization and transport need not necessarily occur in a uniform proximal distal manner. PMID- 12589679 TI - Role of the two type II myosins, Myo2 and Myp2, in cytokinetic actomyosin ring formation and function in fission yeast. AB - The formation and contraction of a cytokinetic actomyosin ring (CAR) is essential for the execution of cytokinesis in fission yeast. Unlike most organisms in which its composition has been investigated, the fission yeast CAR contains two type II myosins encoded by the genes myo2(+) and myp2(+). myo2(+) is an essential gene whilst myp2(+) is dispensable under normal growth conditions. Myo2 is hence the major contractile protein of the CAR whilst Myp2 plays a more subtle and, as yet, incompletely documented role. Using a fission yeast strain in which the chromosomal copy of the myo2(+) gene is fused to the gene encoding green fluorescent protein (GFP), we analysed CAR formation and function in the presence and absence of Myp2. No change in the rate of CAR contraction was observed when Myp2 was absent although the CAR persisted longer in the contracted state and was occasionally observed to split into two discrete rings. This was also observed in myp2Delta cells following actin depolymerisation with latrunculin. CAR contraction in the absence of Myp2 was completely abolished in the presence of elevated levels of chloride ions. Thus, Myp2 appears to contribute to the stability of the CAR, in particular at a late stage of CAR contraction, and to be a component of the signalling pathway that regulates cytokinesis in response to elevated levels of chloride. To determine whether the presence of two type II myosins was a feature of cytokinesis in other fungi that divide by septation, we searched the genomes of two filamentous fungi, Aspergillus fumigatus and Neurospora crassa, for myosin genes. As in fission yeast, both A. fumigatus and N. crassa contained myosins of classes I, II, and V. Unlike fission yeast, both contained a single type II myosin gene that, on the basis of its tail structure, was more reminiscent of Myp2 than Myo2. The significance of these observations to our understanding of septum to formation and cleavage is discussed. PMID- 12589680 TI - Dynamics of the endoplasmic reticulum during early development of Drosophila melanogaster. AB - In this study, we analyze for the first time endoplasmic reticulum (ER) dynamics and organization during oogenesis and embryonic divisions of Drosophila melanogaster using a Protein Disulfide Isomerase (PDI) GFP chimera protein. An accumulation of ER material into the oocyte takes place during the early steps of oogenesis. The compact organization of ER structures undergoes a transition to an expanded reticular network at fertilization. At the syncytial stage, this network connects to the nuclear envelope as each nucleus divides. Time-lapse confocal microscopy on PDI transgenic embryos allowed us to characterize a rapid redistribution of the ER during the mitotic phases. The ER network is massively recruited to the spindle poles in prophase. During metaphase most of the ER remains concentrated at the spindle poles and shortly thereafter forms several layers of membranes along the ruptured nuclear envelope. Later, during telophase an accumulation of ER material occurs at the spindle equator. We also analyzed the subcellular organization of the ER network at the ultrastructural level, allowing us to corroborate the results from confocal microscopy studies. This dynamic redistribution of ER suggests an unexpected regulatory function for this organelle during mitosis. PMID- 12589681 TI - 3-D in vitro model of early skeletal muscle development. AB - An understanding of the mechanical and mechano-molecular responses that occur during the differentiation of mouse C2C12 [corrected] myoblasts in 3-D culture is critical for understanding growth, which is important for progress towards producing a tissue-engineered muscle construct. We have established the main differences in force generation between skeletal myoblasts, dermal fibroblasts, and smooth muscle cells in a 3-D culture model in which cells contract a collagen gel construct. This model was developed to provide a reproducible 3-D muscle organoid in which differences in force generation could be measured, as the skeletal myoblasts fused to form myotubes within a collagen gel. Maintenance of the 3-D culture under sustained uni-axial tension, was found to promote fusion of myoblasts to form aligned multi-nucleate myotubes. Gene expression of both Insulin Like Growth Factor (IGF-1 Ea) and an isoform of IGF-1 Ea, Mechano-growth factor (IGF-1 Eb, also termed MGF), was monitored in this differentiating collagen construct over the time course of fusion and maturation (0-7 days). This identified a transient surge in both IGF-1 and MGF expression on day 3 of the developing construct. This peak of IGF-1 and MGF expression, just prior to differentiation, was consistent with the idea that IGF-1 stimulates differentiation through a Myogenin pathway [Florini et al., 1991: Mol. Endocrinol. 5:718-724]. MGF gene expression was increased 77-fold on day 3, compared to a 36-fold increase with IGF-1 on day 3. This indicates an important role for MGF in either differentiation or, more likely, a response to mechanical or tensional cues. PMID- 12589682 TI - Dynamic properties of APC-decorated microtubules in living cells. AB - The adenomatous polyposis coli (APC) tumour suppressor protein is a component of the Wnt signalling pathway in which it plays a major role in controlling nuclear accumulation of beta-catenin and hence in the modulation of beta-catenin regulated gene transcription. APC also associates with microtubules at the ends of cytoplasmic extensions in epithelial cells, a distribution that can be reproduced in COS cells ectopically expressing APC. To examine the effect of APC on microtubule properties, we monitored directly the behaviour of APC and of APC decorated microtubules by time-lapse imaging of cytoplasmic extensions in live COS cells expressing APC tagged with a green fluorescent protein. On the proximal part of microtubules, APC was visualised as particulate material moving unidirectionally towards the plus end of microtubules. The distal parts of microtubules were uniformly decorated by APC and were animated by a motile behaviour in the form of aperiodic bending. This behaviour is likely to be the consequence of compression forces acting on microtubules encountering obstacles while elongating. The majority of APC-decorated microtubules in transfected COS cells was sensitive to depolymerisation by nocodazole, but they contained detyrosinated and acetylated alpha-tubulin, suggesting a reduction in the rate of subunit exchange at their growing end. Taken together, these results demonstrate that microtubule domains uniformly decorated by APC display dynamic and motile properties that may be significant for the postulated role of APC in targeting microtubules to specialised membrane sites. PMID- 12589683 TI - Smad3 mediates the TGF-beta-induced contraction of type I collagen gels by mouse embryo fibroblasts. AB - TGF-beta signals through TGF-beta receptors and Smad proteins. TGF-beta also augments fibroblast-mediated collagen gel contraction, an in vitro model of connective tissue remodeling. To investigate the importance of Smad2 or Smad3 in this augmentation process, embryo-derived fibroblasts from mice lacking expression of Smad2 or Smad3 genes were cast into native type I collagen gels. Fibroblast-populated gels were then released into 0.2% FCS-DMEM alone or with recombinant human TGF-beta1, beta2, beta3, or recombinant rat PDGF-BB. Gel contraction was determined using an image analyzer. All three isoforms of TGF beta significantly augmented contraction of collagen gels mediated by fibroblasts with genotypes of Smad2 knockout (S2KO), Smad2 wildtype (S2WT), and Smad3 wildtype (S3WT), but not Smad3 knockout (S3KO) mice. PDGF-BB augmented collagen gel contraction by all fibroblast types. These results suggest that expression of Smad3 but not Smad2 may be critical in TGF-beta augmentation of fibroblast mediated collagen gel contraction. Thus, the Smad3 gene could be a target for blocking contraction of fibrotic tissue induced by TGF-beta. PMID- 12589684 TI - ALP and MLP distribution during myofibrillogenesis in cultured cardiomyocytes. AB - The Z-line is a multifunctional macromolecular complex that anchors sarcomeric actin filaments, mediates interactions with intermediate filaments and costameres, and recruits signaling molecules. Antiparallel alpha-actinin homodimers, present at Z-lines, cross-link overlapping actin filaments and also bind other cytoskeletal and signaling elements. Two LIM domain containing proteins, alpha-actinin associated LIM protein (ALP) and muscle LIM protein (MLP), interact with alpha-actinin, distribute in vivo to Z-lines or costameres, respectively, and, when absent, are associated with heart disease. Here we describe the behavior of ALP and MLP during myofibrillogenesis in cultured embryonic chick cardiomyocytes. As myofibrils develop, ALP and MLP are observed in distinct distribution patterns in the cell. ALP is coincident with alpha actinin from the first stage of myofibrillogenesis and co-distributes with alpha actinin to Z-lines and intercalated discs in mature myofibrils. Interestingly, we also demonstrate using ALP-GFP transfection experiments and an in vitro binding assay that the ALP-alpha-actinin binding interaction is not required to target ALP to the Z-line. In contrast, MLP localization is not co-incident with that of alpha-actinin until late stages of myofibrillogenesis; however, it is present in premyofibrils and nascent myofibrils prior to the incorporation of other costameric components such as vinculin, vimentin, or desmin. Our observations support the view that ALP function is required specifically at actin anchorage sites. The subcellular distribution pattern of MLP during myofibrillogenesis suggests that it functions during differentiation prior to the establishment of costameres. PMID- 12589687 TI - Reorganization of monoaminergic systems in the earthworm, Eisenia fetida, following brain extirpation. AB - The present study describes the major aspects of how monoaminergic (serotonin, dopamine) systems change in the course of regeneration of the brain in the earthworm (Eisenia fetida), investigated by immunocytochemistry, HPLC assay, and ligand binding. Following brain extirpation, the total regeneration time is about 80 days at 10 degrees C. On the 3rd postoperative day serotonin, and on the 11th postoperative day tyrosine hydroxylase-immunoreactive neurons can be observed in the wound tissue. Thereafter the number of the immunoreactive cells increases gradually, and by the 76th-80th postoperative days all serotonin- and tyrosine hydroxylase-immunopositive neurons can be found in their final positions, similarly to those observed in the intact brain. Labeled neurons located in the dorsal part of the regenerated brain appear earlier than the cells in lateral and ventral positions. Both serotonin- and tyrosine hydroxylase-immunoreactive neurons of the newly formed brain seem to originate from undifferentiated neuroblasts situated within and around the ventral ganglia and the pleura. Dopaminergic (tyrosine hydroxylase-immunoreactive) elements may additionally derive from the proliferation of neurons localized in the subesophageal ganglion and the pharyngeal nerve plexus. Following brain extirpation, both serotonin and dopamine levels, assayed by HPLC, first increase in the subesophageal ganglion; by the 25th day of regeneration, the monoamine content decreases in it and increases in the brain. Hence it is suggested that monoamines are at least partly transported from this ganglion to the regenerating brain. At the same time, (3)H LSD binding can be detected in the regenerating brain from the 3rd postoperative day, showing a continuous increase until the 80th postoperative day, suggesting a guiding role of postsynaptic elements in the monoaminergic reinnervation of the newly formed brain. PMID- 12589686 TI - Putative histidin-rich proteins in the epidermis of lizards. AB - In the stratum granulosum of mammalian epidermis, histidin-rich proteins (filaggrins) determine keratin clumping and matrix formation into terminal keratinocytes of the stratum corneum. The nature of matrix, interkeratin proteins in the epidermis of nonmammalian vertebrates, and in particular in that of reptilian, mammalian progenitors are unknown. The present biochemical study is the first to address this problem. During a specific period of the renewal phase of the epidermis of lizards and during epidermal regeneration, keratohyalin-like granules are formed, at which time they take up tritiated histidine. The latter also accumulate in cells of the alpha-keratin layer (soft keratin). This pattern of histidine incorporation resembles that seen in keratohyalin granules of the stratum granulosum of mammalian epidermis. After injection of tritiated histidine, we have analysed the distribution of the radioactivity by histoautoradiography and electrophoretic gel autoradiography of epidermal proteins. Extraction and electrophoretic separation of interfilamentous matrix proteins from regenerating epidermis 3-48 hours post-injection reveals the appearance of protein bands at 65-70, 55-58, 40-43, 30-33, 25-27, and 20-22 kDa. Much weaker bands were seen at 100, 140-160, and 200 kDa. A weak band at 20-22 kDa or no bands at all are seen in the normal epidermis in resting phase and in the dermis. In regenerating epidermis at 22 and 48 hours post-injection, little variation in bands is detectable, but low molecular weight bands tend to increase slightly, suggesting metabolic turnover. Using anti-filaggrin antibodies against rat, human, or mouse filaggrins, some cross-reactivity was seen with more reactive bands at 40-42 and 33 kDa, but it was reduced or absent at 140, 95-100, 65-70, 50-55, and 25 kDa. This suggests that different intermediate degradative proteins of lizard epidermis may share some epitopes with mammalian filaggrins and are different from keratins with molecular weight ranging from 40 to 65-68 kDa. The immunocytochemical observation confirms that a weak filaggrin-like immunoreactivity characterizes differentiating alpha-keratogenic layers in normal and regenerating tail. A weak filaggrin labeling is discernable in small keratohyalin-like granules but is absent from the larger granules and from mature keratinocytes. The present results indicate, for the first time, that histidine rich proteins are involved in the process of alpha-keratinization in reptilian epidermis. The cationic, interkeratin matrix proteins implicated may be fundamentally similar in both theropsid-derived and sauropsid amniotes. PMID- 12589688 TI - Entrainment of circadian locomotor activity rhythm of the nocturnal field mouse Mus booduga using daily injections of melatonin. AB - In this paper, we report the effects of daily injections of melatonin on the locomotor activity rhythm of the nocturnal field mouse Mus booduga. The locomotor activity rhythm of 45 animals was first monitored in constant darkness (DD) of the laboratory for about 15 days. The animals were then divided into three groups (experimental, vehicle-treated control, and the nontreated control groups) and subjected to three different treatments. The animals from the experimental group (n=19) were administered daily a single subcutaneous (s.c.) injection of melatonin (1 mg/kg) for about 45 days. The vehicle treated controls (n=13) were administered daily injections of 50% dimethyl sulfoxide (DMSO) for about 45 days, and the nontreated controls (n=13) were handled similar to the other two groups without being administered injections. Following the treatments, the animals were maintained in DD for about 20 days, after which the experiments were terminated. A significantly larger percentage of animals from the experimental group either entrained or showed phase control to daily treatments, compared to the animals from the two control groups. These results suggest that externally administered melatonin can influence the phase of the circadian locomotor activity rhythm of M. booduga. The fact that none of the nontreated controls showed any sign of phase control to daily handling, clearly demonstrates that the entrainment or phase control in the melatonin treated group of animals is caused by melatonin alone and not due to handling. PMID- 12589689 TI - Early photoperiod history and short-day responsiveness in Siberian hamsters. AB - Siberian hamsters exhibit seasonal, photoperiod influenced cycles of reproductive activity, body size, pelage characteristics, and thermoregulatory behavior. Laboratory populations generally exhibit inter-individual variability in expression of photoperiod responsiveness, with a subset of individuals that fail to show the species typical responses to short photoperiod. This variability is partly explained by a genetic component, as it has been possible to increase the number of short-day nonresponders by artificial selection. Responsiveness to short photoperiod is also substantially influenced by photoperiod history in this species; hamsters that have been raised under long (16L) or very long (18L) day lengths are less likely to exhibit winter-type responses to short days as compared to hamsters raised under an intermediate (14L) day length. In the present experiment, we examined effects of age and early photoperiod history in a strain of Siberian hamsters that had been selected for short-day nonresponsiveness. Hamsters transferred into short photoperiod on the day of birth were uniform in exhibiting winter-type responses. However, hamsters raised until 25 days of age in either continuous illumination or in 16L exhibited variation in responsiveness when subsequently moved into short photoperiod. We conclude that virtually all hamsters of the short-day nonresponsive strain are born responsive to short days. Subsequent development of resistance to potential short day effects is dependent on age and/or photoperiod history. PMID- 12589690 TI - Effects of androgens on sex differentiation of the urodele Pleurodeles waltl. AB - In nonmammalian vertebrates, steroids have been hypothesized to induce somatic sex differentiation, since manipulations of the steroidal environment of gonads have led to various degrees of sex reversal. Whereas the critical role of estrogens in ovarian differentiation is well documented, studies on androgens have produced a perplexing variety of results depending upon species variations and nature of androgens used. In this way, testosterone induces masculinization of females in some species but provokes paradoxical feminization of males in many other species such as the urodelan Pleurodeles waltl. In reptiles this phenomenon could be interpreted by conversion of exogenous testosterone to estradiol by aromatase. Treatments of Pleurodeles larvae with nonaromatizable androgens bring support to this hypothesis and suggest a role of androgens in sex differentiation. Dihydrotestosterone (DHT) could not induce the paradoxical feminization of ZZ larvae. In addition, DHT as well as 11beta-hydroxy androstenedione could drive a functional male differentiation of ZW larvae. Moreover, other 5alpha reduced androgens also induced sex reversal of female larvae. Yet, the 5alpha reductase inhibitor CGP 53133 and antiandrogens such as flutamide or cyproterone acetate did not exert any effect on male sex differentiation of ZZ larvae. Though the precise role of androgens is still unknown, especially for 11-oxygenated androgens, our results suggest an implication in male sex differentiation. In this way, testosterone could play a pivotal role in being metabolized either into other androgens during testis differentiation or into estradiol during ovarian differentiation. PMID- 12589691 TI - Development of rhythmic melatonin secretion from the pineal gland of embryonic mummichog (Fundulus heteroclitus). AB - The pineal gland of vertebrates produces and secretes the hormone melatonin in response to changes in the light-dark cycle, with high production at night and low production during the day. Melatonin is thought to play an important role in synchronizing daily and/or seasonal physiological, behavioral, and developmental rhythms in vertebrates. In this study, the functional development of the pineal melatonin-generating system was examined in the mummichog, Fundulus heteroclitus, an euryhaline teleost. In this species, the pineal gland contains an endogenous oscillator, ultimately responsible for timing the melatonin rhythm. Oocytes from gravid females were collected and fertilized in vitro from sperm collected from mature males. Skull caps containing attached pineal glands were obtained from F. heteroclitus embryos at different embryonic stages and placed in static or perfusion culture under various photoperiodic regimes. Rhythmic melatonin secretion from pineal glands of embryonic F. heteroclitus embryos exposed to a 12L:12D cycle in static culture was observed at five days post-fertilization. The ontogeny of circadian-controlled melatonin production from F. heteroclitus pineal glands exposed to constant darkness for five days was also seen at day five post fertilization. These data show that early development of the pineal melatonin generating system in this teleost occurs prior to hatching. Pre-hatching development of the melatonin-generating system may confer some selective advantage in this species in its interactions with the environment. PMID- 12589692 TI - Prolonged response to calling songs by the L3 auditory interneuron in female crickets (Acheta domesticus): intracellular evaluation. AB - The L3 auditory interneuron in female Acheta domesticus, produces two different responses to the male calling song: an immediate response and a prolonged response. The prolonged response exhibited spiking activity and a correlated prolonged depolarization, both of which are clearly seen in intracellular recordings. The morphology revealed by intracellular staining was clearly the L3 neuron. The amplitude of the prolonged depolarization associated with the prolonged response increased with increases in sound intensity, resulting in increased spiking rates. Both depolarization and sound presentation increased the spiking rate and the slope of pre-potentials (thus leading to spiking threshold more quickly). Injecting hyperpolarizing current had the expected opposite effect. The effects of positive current injection and sound presentation were additive, resulting in spiking rates that were approximately double the rates in response to sound alone. Short postsynaptic potentials (PSPs), whose duration ranged from 15-60 ms, which may lead to action potentials were also observed in all recordings and summated with the prolonged depolarization, increasing the probability of spiking. PMID- 12589693 TI - Prolonged response to calling songs by the L3 auditory interneuron in female crickets (Acheta domesticus): possible roles in regulating phonotactic threshold and selectiveness for call carrier frequency. AB - L3, an auditory interneuron in the prothoracic ganglion of female crickets (Acheta domesticus) exhibited two kinds of responses to models of the male's calling song (CS): a previously described, phasically encoded immediate response; a more tonically encoded prolonged response. The onset of the prolonged response required 3-8 sec of stimulation to reach its maximum spiking rate and 6-20 sec to decay once the calling song ceased. It did not encode the syllables of the chirp. The prolonged response was sharply selective for the 4-5 kHz carrier frequency of the male's calling songs and its threshold tuning matched the threshold tuning of phonotaxis, while the immediate response of the same neuron was broadly tuned to a wide range of carrier frequencies. The thresholds for the prolonged response covaried with the changing phonotactic thresholds of 2- and 5-day-old females. Treatment of females with juvenile hormone reduced the thresholds for both phonotaxis and the prolonged response by equivalent amounts. Of the 3 types of responses to CSs provided by the ascending L1 and L3 auditory interneurons, the threshold for L3's prolonged response, on average, best matched the same females phonotactic threshold. The prolonged response was stimulated by inputs from both ears while L3's immediate response was driven only from its axon-ipsilateral ear. The prolonged response was not selective for either the CS's syllable period or chirp rate. PMID- 12589694 TI - Solvation dynamics and proton transfer in supramolecular assemblies. AB - Water molecules confined in a supramolecular assembly control reactivity and dynamics of biological systems in a unique way. In a confined system, water molecules display an ultraslow component of solvation which is slower than that in bulk water by 2-4 orders of magnitude. The ultraslow component arises mainly from the disruption of the hydrogen-bond network of water and the binding of water molecules to a macromolecule. The ultraslow component of solvation markedly retards polar reactions. Many examples of slow dynamics in complex systems, and their implications in biological and natural processes are discussed. PMID- 12589695 TI - End-grafted polysilane--an approach to single polymer science. AB - This Account describes our approach to single polymer science in which we use end grafted polysilane. Single polymer science, by our definition, involves isolating and observing a single polymer chain and extracting its physical properties at the single molecule level. For this purpose, we developed the end-graft technique, which is a method for fixing a polymer terminus on a solid surface by forming a chemical bond between them. We applied this technique to polysilane, a semiconducting polymer with a one-dimensional silicon-catenated backbone. Our recent experimental achievements are reviewed, which include direct observations of polysilane single-molecular structures by atomic force microscopy, the formation of unique supramolecular structures, and the observation of temperature and solvent-dependent UV absorption spectra that are characteristic of isolated polysilane chains end-grafted on a solid surface. PMID- 12589696 TI - "Design" in solid-state chemistry based on phase homologies. The concept of structural evolution and the new megaseries A(m)[M(1+l)Se(2+l)]2m[M(2l+n)Se(2+3l+n)]. AB - An attractive way to advance the design and prediction of new compositions and structures of solid-state compounds is to identify large homologies that are amenable to chemical control. Homologies allow the systematization of phases under a common conceptual umbrella and have the potential to capture the vast majority of existing and possible compounds. We have demonstrated this in the broad system A/M'/M''/Se (A = K, Rb, Cs, Sr, Ba; M' = Pb, Sn; M'' = Sb, Bi) by identifying the homologous "superseries" A(m)[M(1+l)Se(2+l)]2m[M(2l+n)Se(2+3l+n)]. This series generates a large variety of compounds with predictable composition and structure. All the compounds contain fundamental building units representing different fragments of the NaCl type lattice. The three independent integers l, m, and n determine width, height, and shape of the building units and therefore cause structural evolution of the homologous series in three different dimensions. On the basis of this general formula one can design, in a modular fashion, new compounds that fit the structural evolution of the superseries, predicting simultaneously their structure and composition. Several new phases have been discovered with this approach. Here we give an overview of the character and predictive properties of the superseries and propose the classification of phases into homologies (when possible) which could serve as devices to predict new members. PMID- 12589697 TI - Chlorine kinetic isotope effects on enzymatic dehalogenations. AB - Enzymatic dehalogenation reactions are important for the bioremediation of the environment because of the increasing anthropogenic pollution with halogen containing organic compounds. Chlorine kinetic isotope effects have been measured for four hydrolytic dehalogenases. On the basis of these isotope effects, several details of the mechanisms of the enzymatic dehalogenation reactions have been revealed. PMID- 12589698 TI - Paraherquamides, brevianamides, and asperparalines: laboratory synthesis and biosynthesis. An interim report. AB - Studies from our laboratories on the paraherquamide, brevianamide, and asperparaline families of natural products are reviewed. It has been proposed that the unique core ring system that is common to this family of compounds arises by a biological intramolecular Diels-Alder cycloaddition reaction. Key biosynthetic studies are described, along with classical synthetic approaches as well as those inspired by Nature for the synthesis of these interesting molecules. PMID- 12589699 TI - Isotope effects in C-H bond activation reactions by transition metals. AB - The activation of alkane C-H bonds by oxidative addition and its reverse reaction, reductive elimination, are believed to occur via transient sigma-alkane complexes. This Account summarizes how isotope effects can be used to probe the nature of these intermediates and points out some pitfalls in interpreting kinetic data. Comparisons are made with arene C-H activation and other activation systems. PMID- 12589700 TI - Nonoctahedral structures. AB - There is always a special reason when six-coordinate molecules or complexes are nonoctahedral. The Jahn-Teller distortion is the longest known example. Also well understood now is the steric activity or nonactivity of a nonbonding electron pair, for example XeF(6). In the past few years, it became obvious that six coordinate d(0) and d(1) complexes with sigma only ligands such as Mo(CH3)(6) are far from octahedral; rather, they are trigonal prismatic or even C(3v) distorted trigonal prismatic. This phenomenon can be explained with simple molecular orbital or valence bond models. PMID- 12589701 TI - Vertebrate tankyrase domain structure and sterile alpha motif (SAM)-mediated multimerization. AB - Tankyrases 1 and 2 are two highly related poly(ADP-ribose) polymerases that interact with a variety of cytoplasmic and nuclear proteins. Both proteins have been implicated in telomere length regulation, insulin signalling and centrosome function. To learn more about their mode of action, we have isolated the chicken tankyrase homologues and examined their interaction partners and subcellular location. Cross-species sequence comparison indicated that tankyrase domain structure is highly conserved and supports division of the ankyrin domain into five subdomains, which are each separated by a highly conserved LLEAAR/K motif. Glutathione S-transferase pull-down experiments demonstrated that the ankyrin domains of both proteins interact with chicken telomere repeat factor 1 (TRF1). Analysis of total cellular and nuclear proteins revealed that cells contain approximately twice as much tankyrase 1 as tankyrase 2. Although > or = 90% of each protein is present in the cytoplasm, both tankyrase 1 and 2 were detected in the nucleus. The nuclear location together with its ability to interact with TRF1, point to tankyrase 2 having a telomeric function. Yeast two-hybrid and cross-linking experiments show that both tankyrases can multimerize through their sterile-alpha motif domains. These results indicate that tankyrases may be master scaffolding proteins, capable of regulating assembly of large protein complexes. PMID- 12589702 TI - Supplementation with fish oil affects the association between very long-chain n-3 polyunsaturated fatty acids in serum non-esterified fatty acids and soluble vascular cell adhesion molecule-1. AB - We have investigated the effect of fish oil supplementation on the association between serum non-esterified fatty acid (NEFA) pattern and atherosclerotic activity. We studied correlations between serum non-esterified very long-chain eicosapentaenoic (EPA), docosahexaenoic acid (DHA) and arachidonic acid (AA) and biochemical markers of endothelial activation before and after 18-months intervention with fish oil supplementation. The fish oil supplementation consisted of 2.4 g of EPA and DHA per day, with corn oil as placebo. Elderly men ( n =171) with high risk for coronary heart disease were divided into four intervention groups in a factorial design: fish oil supplementation ( n =44), dietary intervention ( n =42), fish oil supplementation+dietary intervention ( n =47) or placebo ( n =38). The composition of fasting NEFA was analysed before and after intervention by GLC. Circulating endothelial markers were analysed by ELISA. A statistically significant positive correlation between the change in serum non-esterified DHA and soluble vascular cell adhesion molecule-1 (sVCAM-1) was found in the pooled group that received fish oil supplementation ( n =91; Spearman's correlation coefficient r =0.24, P =0.02). No such correlation was found in the pooled group without fish oil supplementation ( n =80). Furthermore, there was a significant negative correlation between the change in serum non esterified EPA and the relative change in sVCAM-1 in the group that did not receive fish oil supplementation ( r =-0.34, P =0.002). No such correlation was found in the group with fish oil supplementation. We conclude that large increase in serum non-esterified EPA and DHA, which can only be attained by supplementation, might increase inflammation in vascular endothelium. A moderate dietary increase in fish oil intake may, however, have an effect on decreasing inflammatory markers. PMID- 12589703 TI - Measuring whole-body actin/myosin protein breakdown in mice using a primed constant stable isotope-infusion protocol. AB - To measure actin/myosin protein breakdown, the 24 h excretion of N (tau) methylhistidine (3MH) is used. However, in mice, this method is invalid. Therefore we have developed a liquid chromatography-MS technique to measure the tracer/tracee ratio and concentration of 3MH in plasma, enabling an in vivo primed constant infusion protocol with a deuterated stable isotope of 3MH. We tested this model by giving a primed constant infusion of L-[3-methyl (2)H(3)]histidine, L-[phenyl-(2)H(5)]phenylalanine and L-[phenyl-(2)H(2)]tyrosine to three anaesthetized experimental groups: mice receiving saline intraperitoneally (i.p.) (CON), mice receiving saline i.p. and starved for 9 h (STA), and mice receiving lipopolysaccharide i.p. and starved for 9 h (STA+LPS). The contribution of myofibrillar to total protein breakdown was significantly lower in the STA group than the CON group (30+/-4% and 54+/-14% respectively; P <0.05), and was significantly higher in the STA+LPS group than the STA group (52+/-7% and 30+/-4% respectively; P <0.05). Whole-body myofibrillar protein breakdown, total protein breakdown, protein synthesis and net protein breakdown were not different between the groups. We conclude that this in vivo primed constant stable isotope-infusion protocol can give valuable information about the role of actin/myosin protein breakdown in mice. PMID- 12589704 TI - Blood flow and muscle metabolism in chronic fatigue syndrome. AB - The purpose of this study was to determine if chronic fatigue syndrome (CFS) is associated with reduced blood flow and oxidative delivery to skeletal muscle. Patients with CFS according to CDC (Center for Disease Control) criteria ( n =19) were compared with normal sedentary subjects ( n =11). Muscle blood flow was measured with Doppler ultrasound after cuff ischaemia and exercise. Muscle oxygen delivery was measured as the rate of post-exercise and post-ischaemic oxygen-haem resaturation. Oxygen-haem resaturation was measured in the medial gastrocnemius muscle using continuous wavelength near-IR spectroscopy. Muscle metabolism was measured using (31)P magnetic resonance spectroscopy. CFS patients and controls were not different in the peak blood flow after cuff ischaemia, the rate of recovery of phosphocreatine after submaximal exercise, and the rate of recovery of oxygen saturation after cuff ischaemia. In conclusion, CFS patients showed no deficit in blood flow or oxidative metabolism. This suggests that CFS symptoms do not require abnormal peripheral function. PMID- 12589705 TI - Expression of Toll-like receptor 4 is associated with enteroviral replication in human myocarditis. AB - Previous studies have demonstrated that inflammatory cytokine expression associated with enteroviral (EV) infection may play an important role in human myocarditis. However, the mechanism of the host immune response against viral pathogens has not been fully understood. The aim of the present study was to determine whether Toll-like receptor 4 (TLR4) and EV RNA are present in human myocarditis. Endomyocardial biopsy samples were obtained from 44 patients with myocarditis and five controls. Levels of plus- and minus-strand EV RNAs and TLR4 mRNA were measured by real-time reverse transcriptase-PCR. Immunohistochemical analysis was performed to identify the cellular source of TLR4 and the EV capsid protein VP1. EV RNA was present in 21 patients with myocarditis and these patients were defined as having either active viral replication ( n =15) or latent viral persistence ( n =6). Neither strand of EV RNA was detected in controls. TLR4 mRNA expression levels were higher in myocarditis patients than in controls (TLR4/glyceraldehyde-3-phosphate dehydrogenase ratio 1.48+/-0.17 compared with 0.08+/-0.06, P <0.001). A positive correlation was found between EV RNA and TLR4 levels (plus-strand vs TLR4: r =0.66, P <0.001; minus-strand vs TLR4: r =0.48, P <0.001). TLR4 immunostaining was observed in infiltrating cells and myocytes in patients with myocarditis. The EV capsid protein VP1 was also found in myocytes. The myocarditis group with EV replication and high levels of TLR4 showed significantly lower systolic function. The present study has shown that increased expression of TLR4 is associated with EV replication and that these RNA levels are related to cardiac dysfunction in human myocarditis. PMID- 12589706 TI - Effect of caspase cleavage-site phosphorylation on proteolysis. AB - Caspases are important mediators of apoptotic cell death. Several cellular protein substrates of caspases contain potential phosphorylation site(s) at the cleavage-site region, and some of these sites have been verified to be phosphorylated. Since phosphorylation may affect substantially the substrate susceptibility towards proteolysis, phosphorylated, non-phosphorylated and substituted oligopeptides representing such cleavage sites were studied as substrates of apoptotic caspases 3, 7 and 8. Peptides containing phosphorylated serine residues at P4 and P1' positions were found to be substantially less susceptible towards proteolysis as compared with the serine-containing analogues, while phosphoserine at P3 did not have a substantial effect. P1 serine as well as P1-phosphorylated, serine-containing analogues of an oligopeptide representing the poly(ADP-ribose) polymerase cleavage site of caspase-3 were not hydrolysed by any of these enzymes, whereas the P1 aspartate-containing peptides were efficiently hydrolysed. These findings were interpreted with the aid of molecular modelling. Our results suggest that cleavage-site phosphorylation in certain positions could be disadvantageous or detrimental with respect to cleavability by caspases. Cleavage-site phosphorylation may therefore provide a regulatory mechanism to protect substrates from caspase-mediated degradation. PMID- 12589708 TI - Partially esterified oligogalacturonides are the preferred substrates for pectin methylesterase of Aspergillus niger. AB - Investigations on the mode of action of Aspergillus niger pectin methylesterase (PME) towards differently C(6)- and C(1)-substituted oligogalacturonides (oligoGal p A) are described. De-esterification of methyl-esterified (un)saturated oligoGal p A proceeds via a specific pattern, depending on the degree of polymerization. Initially, a first methyl ester of the oligomer is hydrolysed, resulting in one free carboxyl group. Subsequently, this first product is preferred as a substrate and is de-esterified for a second time. This product is then accumulated and hereafter de-esterified further to the final product, i.e. oligoGal p A containing one methyl ester located at the non reducing end residue for both saturated and unsaturated oligoGal p A, as found by post-source decay matrix-assisted laser-desorption/ionization-time-of-flight MS. The saturated hexamer is an exception to this: three methyl esters are removed very rapidly, instead of two methyl esters. When unsaturated oligoGal p A were used, the formation of the end product differed slightly, suggesting that the unsaturated bond at the non-reducing end influences the de-esterification process. In vivo, PME prefers methyl esters, but the enzyme appeared to be tolerant for other C(6)- and C(1)-substituents. Changing the type of ester (ethyl esterification) or addition of a methyl glycoside (C(1)) only reduced the activity or had no effect respectively. The specific product pattern was identical for all methyl- and ethyl-esterified oligoGal p A and methyl glycosidated oligoGal p A, which strongly indicates that one or perhaps two non esterified oligoGal p A are preferred in the active-site cleft. PMID- 12589707 TI - Role for AMP-activated protein kinase in glucose-stimulated insulin secretion and preproinsulin gene expression. AB - AMP-activated protein kinase (AMPK) has recently been implicated in the control of preproinsulin gene expression in pancreatic islet beta-cells [da Silva Xavier, Leclerc, Salt, Doiron, Hardie, Kahn and Rutter (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 4023-4028]. Using pharmacological and molecular strategies to regulate AMPK activity in rat islets and clonal MIN6 beta-cells, we show here that the effects of AMPK are exerted largely upstream of insulin release. Thus forced increases in AMPK activity achieved pharmacologically with 5-amino-4 imidazolecarboxamide riboside (AICAR), or by adenoviral overexpression of a truncated, constitutively active form of the enzyme (AMPK alpha 1.T(172)D), blocked glucose-stimulated insulin secretion. In MIN6 cells, activation of AMPK suppressed glucose metabolism, as assessed by changes in total, cytosolic or mitochondrial [ATP] and NAD(P)H, and reduced increases in intracellular [Ca(2+)] caused by either glucose or tolbutamide. By contrast, inactivation of AMPK by expression of a dominant-negative form of the enzyme mutated in the catalytic site (AMPK alpha 1.D(157)A) did not affect glucose-stimulated increases in [ATP], NAD(P)H or intracellular [Ca(2+)], but led to the unregulated release of insulin. These results indicate that inhibition of AMPK by glucose is essential for the activation of insulin secretion by the sugar, and may contribute to the transcriptional stimulation of the preproinsulin gene. Modulation of AMPK activity in the beta-cell may thus represent a novel therapeutic strategy for the treatment of type 2 diabetes mellitus. PMID- 12589709 TI - Cigarette smoking, health status, socio-economic status and access to health care in diabetes mellitus: a cross-sectional survey. AB - BACKGROUND: In diabetes mellitus, cigarette smoking is associated with increased risk of cardiovascular mortality and microvascular complications. We evaluated cigarette smoking in people with diabetes mellitus in a socio-economically deprived area. METHODS: We carried out a cross-sectional survey of people registered with diabetes mellitus at 29 general practices in inner London. Responses were analysed for 1,899 (64%) respondents out of 2,983 eligible. RESULTS: There were 1,899 respondents of whom 968 (51%) had never smoked, 296 (16%) were current smokers and 582 (31%) were ex-smokers. Smoking was more frequent in white Europeans (men 22%, women 20%), than in African Caribbeans (men 15%, women 10%) or Africans (men 8%, women 2%). Smoking prevalence decreased with age. Smokers were more likely to be living in rented accommodation (odds ratio, OR 2.02, 95% confidence interval 1.48 to 2.74). After adjusting for confounding, current smokers had lower SF-36 scores than subjects who had never smoked (mean difference in physical functioning score -5.6, 95% confidence interval -10.0 to 1.2; general health -6.1, -9.7 to -2.5). Current smokers were less likely to have attended a hospital diabetic clinic in the last year (OR 0.59, 0.44 to 0.79), and their hypertension was less likely to be treated (OR 0.47, 0.30 to 0.74). CONCLUSIONS: Compared with non-smokers, smokers had lower socio-economic status and worse health status, but were less likely to be referred to hospital or treated for their hypertension. People with diabetes who smoke can be regarded as a vulnerable group who need more intensive support and treatment. PMID- 12589710 TI - Relative value to surgical patients and anesthesia providers of selected anesthesia related outcomes. AB - BACKGROUND: Anesthesia side effects are almost inevitable in most situations. In order to optimize the anesthetic experience from the patient's viewpoint, it makes intuitive sense to attempt to avoid the side effects that the patient fears the most. METHODS: We obtained rankings and quantitative estimates of the relative importance of nine experiences that commonly occur after anesthesia and surgery from 109 patients prior to their surgery and from 30 anesthesiologists. RESULTS: Pain was the most important thing to avoid, and subjects allocated a median of 25 dollars of an imaginary 100 dollars to avoiding it. Next came vomiting (20 dollars), nausea (10 dollars), urinary retention (5 dollars) , myalgia (2 dollars) and pruritus (2 dollars) . Avoiding blood transfusion, an awake anesthetic technique or postoperative somnolence was not given value by the group as a whole. Anesthesiologists valued perioperative experiences in the same way as patients. CONCLUSIONS: Our results are comparable with those of previous studies in the area, and suggest that patients can prioritize the perioperative experiences they wish to avoid during their perioperative care. Such data, if obtained in the appropriate fashion, would enable anesthetic techniques to be compared using decision analysis. PMID- 12589711 TI - Randomized controlled trials in pediatric complementary and alternative medicine: where can they be found? AB - BACKGROUND: The safety and effectiveness of CAM interventions are of great relevance to pediatric health care providers. The objective of this study is to identify sources of reported randomized controlled trials (RCTs) in the field of pediatric complementary and alternative medicine (CAM). METHODS: Reports of RCTs were identified by searching Medline and 12 additional bibliographic databases and by reviewing the reference lists of previously identified pediatric CAM systematic reviews. RESULTS: We identified 908 reports of RCTs that included children under 18 and investigated a CAM therapy. Since 1965, there has been a steady growth in the number of these trials that are being published. The four journals that published the most reported RCTs are The American Journal of Clinical Nutrition, Pediatrics, Journal of Pediatrics, and Lancet. Medline, CAB Health, and Embase were the best database sources for identifying these studies; they indexed 93.2%, 58.4% and 42.2 % respectively of the journals publishing reports of pediatric CAM RCTs. CONCLUSIONS: Those working or interested in the field of pediatric CAM should routinely search Medline, CAB Health and Embase for literature in the field. The four core journals identified above should be included in their collection. PMID- 12589714 TI - 3-d computer visualization and animation of medical images. PMID- 12589712 TI - M6P/IGF2R loss of heterozygosity in head and neck cancer associated with poor patient prognosis. AB - BACKGROUND: The mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF2R) encodes for a multifunctional receptor involved in lysosomal enzyme trafficking, fetal organogenesis, cytotoxic T cell-induced apoptosis and tumor suppression. The purpose of this investigation was to determine if the M6P/IGF2R tumor suppressor gene is mutated in human head and neck cancer, and if allelic loss is associated with poor patient prognosis. METHODS: M6P/IGF2R loss of heterozygosity in locally advanced squamous cell carcinoma of the head and neck was assessed with six different gene-specific nucleotide polymorphisms. The patients studied were enrolled in a phase 3 trial of twice daily radiotherapy with or without concurrent chemotherapy; median follow-up for surviving patients is 76 months. RESULTS: M6P/IGF2R was polymorphic in 64% (56/87) of patients, and 54% (30/56) of the tumors in these informative patients had loss of heterozygosity. M6P/IGF2R loss of heterozygosity was associated with a significantly reduced 5 year relapse-free survival (23% vs. 69%, p = 0.02), locoregional control (34% vs. 75%, p = 0.03) and cause specific survival (29% vs. 75%, p = 0.02) in the patients treated with radiotherapy alone. Concomitant chemotherapy resulted in a better outcome when compared to radiotherapy alone only in those patients whose tumors had M6P/IGF2R loss of heterozygosity. CONCLUSIONS: This study provides the first evidence that M6P/IGF2R loss of heterozygosity predicts for poor therapeutic outcome in patients treated with radiotherapy alone. Our findings also indicate that head and neck cancer patients with M6P/IGF2R allelic loss benefit most from concurrent chemotherapy. PMID- 12589713 TI - Viable nonsense mutants for the essential gene SUP45 of Saccharomyces cerevisiae. AB - BACKGROUND: Termination of protein synthesis in eukaryotes involves at least two polypeptide release factors (eRFs) - eRF1 and eRF3. The highly conserved translation termination factor eRF1 in Saccharomyces cerevisiae is encoded by the essential gene SUP45. RESULTS: We have isolated five sup45-n (n from nonsense) mutations that cause nonsense substitutions in the following amino acid positions of eRF1: Y53 --> UAA, E266 --> UAA, L283 --> UAA, L317 --> UGA, E385 --> UAA. We found that full-length eRF1 protein is present in all mutants, although in decreased amounts. All mutations are situated in a weak termination context. All these sup45-n mutations are viable in different genetic backgrounds, however their viability increases after growth in the absence of wild-type allele. Any of sup45-n mutations result in temperature sensitivity (37 degrees C). Most of the sup45-n mutations lead to decreased spore viability and spores bearing sup45-n mutations are characterized by limited budding after germination leading to formation of microcolonies of 4-20 cells. CONCLUSIONS: Nonsense mutations in the essential gene SUP45 can be isolated in the absence of tRNA nonsense suppressors. PMID- 12589715 TI - Lumbar spine visualisation based on kinematic analysis from videofluoroscopic imaging. AB - Low back pain is a significant problem and its cost is enormous to society. However, diagnosis of the underlying causes remains problematic despite extensive study. Reasons for this arise from the deep-rooted situation of the spine and also from its structural complexity. Clinicians have to mentally convert 2-D image information into a 3-D form to gain a better understanding of structural integrity. Therefore, visualisation and animation may be helpful for understanding, diagnosis and for guiding therapy. Some low back pain originates from mechanical disorders, and study of the spine kinematics may provide an insight into the source of the problem. Digital videofluoroscopy was used in this study to provide 2-D image sequences of the spine in motion, but the images often suffer due to noise, exacerbated by the very low radiation dosage. Thus determining vertebrae position within the image sequence presents a considerable challenge. This paper describes a combination of spine kinematic measurements with a solid model of the human lumbar spine for visualisation of spine motion. Since determination of the spine kinematics provides the foundation and vertebral extraction is at the core, this is discussed in detail. Edge detection is a key feature of segmentation and it is shown that phase congruency performs better than most established methods with the rather low-grade image sequences from fluoroscopy. The Hough transform is then applied to determine the positions of vertebrae in each frame of a motion sequence. In the Hough transform, Fourier descriptors are used to represent the vertebral shapes. The results show that the Hough transform is a very promising technique for vertebral extraction from videofluoroscopic images. A dynamic visualisation package has been developed in order to view the moving lumbar spine from any angle and viewpoint. Wire frame models of the vertebrae were built by using CT images from the Visible Human Project and these models are scaled to match the fluoroscopic image data. For animation, the spinal kinematic data from the motion study is incorporated. PMID- 12589716 TI - Analysis of the TMJ intraarticular space variation: a non-invasive insight during mastication. AB - The analysis of the loading of the temporomandibular joint (TMJ) appears important for the study of joint failure. Reconstruction and animation of the TMJ with real anatomic and kinematic data is currently the only method allowing a non invasive, in vivo, three-dimensional, dynamic, real-time quantitative insight into the relationship between the articulating surfaces of a joint. It consists of the software reconstruction of the joint anatomy from tomography data and the application to it of the corresponding real movement data recorded with a tracking device. Because of the physical properties of the TMJ soft tissues, the reduction of the joint space can indicate compressive loads during function. Analyses of the variation of the TMJ space in ten asymptomatic subjects during unilateral mastication showed a TMJ space that was significantly smaller during closing than during opening (p<0.05) and significantly smaller on the balancing than on the working joint (p<0.05). This occurs especially in the medial part of the posterior slope of the articular eminence at the end of the closing phase (p<0.01). Furthermore, the minimum TMJ space at the end of closing increased from the beginning to the end of food comminution (p<0.01). PMID- 12589717 TI - Relation of jaw sounds and kinematics visualized and quantified using 3-D computer animation. AB - The management of jaw pain or temporomandibular disorders (TMD) has been controversial regarding temporomandibular joint (TMJ) sounds and their implication regarding TMD prognosis. 3-D computer animation was used to visualize and quantify the internal mechanics of natural mandibular motion synchronized with TMJ sounds. Mandibular movements of four TMD patients and two healthy subjects were recorded using CCD cameras and reflective markers. Sounds were recorded with electret microphones. Magnetic resonance imaging was used to create 3-D geometric models. Visualization of the internal anatomy, mandibular condyle and glenoid fossa, revealed that the condyle initially rotated within the fossa and then moved out of the fossa along, and well beyond, the articular eminence. Power in the opening sound recordings after the condyle moved out of the fossa was significantly greater than when the condyle was within the fossa (p<0.001). The louder opening sounds were often classified as TMJ clicks, implying that clicks occur after the condyle moves out of the fossa. The 3-D computer animation should help resolve the implication of TMJ sounds regarding TMD prognosis by providing visualization and quantization of the TMJ internal mechanics during sound production. PMID- 12589718 TI - Graphic-based musculoskeletal model for biomechanical analyses and animation. AB - The ability to combine physiology and engineering analyses with computer sciences has opened the door to the possibility of creating the 'Virtual Human' reality. This paper presents a broad foundation for a full-featured biomechanical simulator for the human musculoskeletal system physiology. This simulation technology unites the expertise in biomechanical analysis and graphic modeling to investigate joint and connective tissue mechanics at the structural level and to visualize the results in both static and animated forms together with the model. Adaptable anatomical models including prosthetic implants and fracture fixation devices and a robust computational infrastructure for static, kinematic, kinetic, and stress analyses under varying boundary and loading conditions are incorporated on a common platform, the VIMS (Virtual Interactive Musculoskeletal System). Within this software system, a manageable database containing long bone dimensions, connective tissue material properties and a library of skeletal joint system functional activities and loading conditions are also available and they can easily be modified, updated and expanded. Application software is also available to allow end-users to perform biomechanical analyses interactively. This paper details the design, capabilities, and features of the VIMS development at Johns Hopkins University, an effort possible only through academic and commercial collaborations. Examples using these models and the computational algorithms in a virtual laboratory environment are used to demonstrate the utility of this unique database and simulation technology. This integrated system will impact on medical education, basic research, device development and application, and clinical patient care related to musculoskeletal diseases, trauma, and rehabilitation. PMID- 12589719 TI - Data representation for joint kinematics simulation of the lower limb within an educational context. AB - Three-dimensional (3D) visualization is becoming increasingly frequent in both qualitative and quantitative biomechanical studies of anatomical structures involving multiple data sources (e.g. morphological data and kinematics data). For many years, this kind of experiment was limited to the use of bi-dimensional images due to a lack of accurate 3D data. However, recent progress in medical imaging and computer graphics has forged new perspectives. Indeed, new techniques allow the development of an interactive interface for the simulation of human motions combining data from both medical imaging (i.e., morphology) and biomechanical studies (i.e., kinematics). Fields of application include medical education, biomechanical research and clinical research. This paper presents an experimental protocol for the development of anatomically realistic joint simulation within a pedagogical context. Results are shown for the lower limb. Extension to other joints is straightforward. This work is part of the Virtual Animation of the Kinematics of the Human project (VAKHUM) (http://www.ulb.ac.be/project/vakhum). PMID- 12589720 TI - Effect of specimen length: are the mechanics of individual motion segments comparable in functional spinal units and multisegment specimens? AB - Functional spinal units (FSUs) are frequently used for in vitro mechanical testing. This approach assumes that the mechanical behavior of the FSUs is equivalent to the mechanics of these segments within the intact spine. The purpose of this study was to determine whether normal spinal mechanics are compromised in FSU preparations. Flexion-extension pure-moment flexibility testing was performed on 13 L2-L5 porcine specimens. The moment-angle relationship of the L3/L4 segment was recorded, and then the multisegment specimens were cut down to L3/L4 FSUs and retested. Comparisons of stiffness, range of motion, and laxity zone were made between conditions. The neutral zone and range of motion parameters were significantly larger in the L3/L4 motion segment compared to the L3/L4 segment tested within the multisegment specimen; the stiffness was not significantly different. These differences were attributed to cutting the supraspinous ligament as this ligament spans several vertebral levels. Flexion mechanical tests performed on FSUs should be interpreted cautiously as the biomechanics of FSUs is altered from normal. Although the choice of spine length depends on the experimental purpose, spinal flexion studies should be performed on multisegment specimens to appropriately represent the anatomical boundary conditions. PMID- 12589721 TI - Umbilical flow distribution to the liver and the ductus venosus in human fetuses during gestation: an anatomy-based mathematical modeling. AB - The partitioning of umbilical vein blood flow between fetal liver and ductus venosus may be an indicator of the fetal well-being, because the goal of the ductus venosus is to supply oxygen and nutrients to heart and brain. Both distribution and blood flow rate of the umbilical vein are functions of the local vascular impedances that, in turn, depend on the anatomical features of the related vessels. In order to investigate the venous blood flows in human fetuses during a normal gestation, a simple lumped parameter mathematical model was developed on the basis of some information achievable by ultrasonographic techniques. Particularly, the diameter and length of umbilical vein and ductus venosus and the volume of the liver were used to derive the vascular impedances. Three different impedance models were adopted for the umbilical vein, the ductus venosus and the hepatic circulation. A linear model described viscous hydraulic dissipations through the umbilical vein, while a quadratic pressure-flow relationship was used for the ductus venosus due to the irregular local hemodynamics at its inlet. Finally, the equivalent impedance of the whole hepatic network was related to the hepatic volume assuming a tree-like, symmetric and self-similar fractal geometry. The hepatic vascular resistances predicted according to the fractal analysis were quite consistent with some experimental measurements in fetal lambs. In agreement with clinical observations, the model predicted blood flows through the ductus venosus and umbilical vein increasing (from about 25 to 75 ml/min and from about 45 to 370 ml/min, respectively) throughout the gestation (20-40 weeks), while the flow fraction shunted via the ductus venosus diminishes (from about 50 to 20%). PMID- 12589722 TI - An implantable device for stimulation of denervated muscles in rats. AB - The purposes of the present study were (1) to develop an implantable device capable of being pre-programmed to generate a protocol of chronic contractions in denervated hind-limb muscles of rats, and (2) to verify the design by implanting the stimulators for five weeks in rats to identify a protocol of stimulation that maintains muscle mass and maximum force in stimulated-denervated extensor digitorum longus (EDL) muscles. This implantable stimulator system did not hinder animal movement or hygiene, and enabled the animals to be housed in regular animal facilities, since neither external equipment nor an externally generated magnetic field was required. The pre-programmable microcontroller allows detailed basic research into the cellular and tissue response to different stimulation protocols. The micropower design of the battery powered device enabled chronic stimulation of denervated EDL muscles for the five weeks of this initial study. Stimulation protocols of 9-11 V pulse amplitude, 0.4 ms bipolar pulse width, 100 Hz, 20 pulses per contraction, and 100 or 300 contractions generated per day maintained muscle mass and maximum force in denervated EDL muscles of rats at values near control values for innervated muscles. PMID- 12589723 TI - A medical needle drive for the study of interstitial implant mechanics. AB - We describe the construction and test performance of a computer-controlled medical needle drive. The drive represents one facet of a larger project whose aim is to investigate experimentally the mechanics of needle introduction in radioactive 'seed' prostate implants, with a view to identifying ways of making incremental improvements in needle placement accuracy. It is capable of mimicking a range of motions imparted to a needle by a clinical practitioner, and of monitoring the compressive force at the needle tip in real time via an in-line load cell. Tests involving driving needles into porcine gelatin samples using a variety of velocity profiles confirm intended operation. The drive will permit us to introduce needles in a controlled and reproducible manner into a realistic prostate phantom currently being designed. PMID- 12589724 TI - The caddisfly Ceraclea fulva and the freshwater sponge Ephydatia fluviatilis: a successful relationship. AB - The association between the aquatic phases of the caddisfly Ceraclea fulva (Trichoptera, Leptoceridae) and the freshwater sponge Ephydatia fluviatilis (Porifera, Spongillidae) has been investigated by means of scanning and transmission electron microscopy (SEM, TEM). Ceraclea fulva habitually feeds on sponges and builds its case by using the siliceous spicules of the sponge, which are arranged side by side, inter-crossed, cemented with silk, and organised in layers. In the newly hatched larva, the case is strengthened exclusively by cemented siliceous spicules, while during growth, the insect adds sponge fragments to it. The fine organisation of the sponge tissues growing on the case proves that the sponge is functional. Inter-spaced, small protrusions, derived from the outermost compact silk layer, form a series of "bridges" enhancing case/sponge adhesion. Tube-case shape varies according to the aquatic developmental phase of the insect: in the mature larva and pupa, this shelter carries larger sponge fragments dorsally. The caddisfly acts as carrier of the sponge, thus facilitating its dispersal and the colonisation of new habitats. This justifies regarding this association as a successful mutualistic relationship, and not as a unilateral parasitic behaviour on the part of the insect. PMID- 12589725 TI - Ovarian follicle ultrastructure in the teleost Synbranchus marmoratus (Bloch, 1795), with special reference to the vitelline envelope development. AB - Synbranchus marmoratus is a protogynous diandric teleost fish widely distributed throughout South America. The aim of this work was to study the ultrastructure of the vitelline envelope and the relationship among oocyte and their follicular cells during oogenesis. During perinucleolar stage, the oocyte and the follicular cells form microvillar processes that project into the perivitelline space. The oocyte secretes a dense and amorphous material, which appears as the first evidence of the vitelline envelope (VE) development. The VE passes from a double to a multilayered structure during oocyte growth. In mature oocytes, the VE reach a mean thickness of 11 microm, having up to 30 layers. Oocyte microvilli are thinner than the follicular ones and were seen in contact with the follicular plasmalema, however we could not find any contact between the follicular microvilli and the oolemma. Before ovulation, microvillar processes retract and the pore canals seem to collapse. An outer electron dense layer occludes the superficial pore and forms a continuous layer. No jelly or adhesive coatings were seen at least in ovulated eggs sampled from ovarian lumen. Follicular cell and oocyte cytological characteristics do not differ from those described in other teleosts species. PMID- 12589726 TI - New findings on sperm ultrastructure of Xenos vesparum (Rossi) (Strepsiptera, Insecta). AB - The systematic position of insect order Strepsiptera is still under debate. It was, therefore, thought of interest to examine the ultrastructure of a strepsipteran in a search for synapomorphies shared with Coleoptera, Diptera, or any other insect order. The fine structure of spermatozoa and the spermatid from Xenos vesparum (Rossi) was re-examined using scanning and transmission electron microscopy and a fixation technique that permits the visualization of the macromolecular organization of the organelles. The spermatozoon was shown to possess several traits that are characteristics of insects in general, such as a 9 + 9 + 2 axoneme, two mitochondrial derivatives containing a crystalline material and two 'zipper lines' present along the sperm tail. Seventeen protofilaments occurred along most of the accessory tubules, which reduced to 16 posteriorly. An acrosome is absent. The neck region contains a prominent centriolar adjunct, which gives rise to two accessory bodies which adhere to the mitochondrial derivatives, and to slender strands of the so-called intertubular material found between the accessory tubules. Of interest is the finding that the glycocalyx consists of prominent filamentous strands, similar to those found in siphonapterans, mecopterans and basal dipterans. PMID- 12589727 TI - Lymphocytes apoptosis: young versus aged and humans versus rats. AB - This paper deals with a comparative study of lymphocyte apoptosis in young versus aged and humans versus rats. Apoptotic rate achieved by the use of different apoptogenic inducers, acting at different cellular levels, and cell surface modifications were analyzed. The results showed that aged human lymphocytes and freshly isolated rat lymphocytes were more prone to undergo apoptosis. Therefore, the same apoptotic signal is recognized by human and rat lymphocytes, but the extent of the answer is related to the species, to the intensity of the apoptotic stimulus and to the metabolic/developmental condition of the cells. Surface modifications (lipids and glycans), typical of apoptosis, were observed. Our data showed that cell surface changes are species and age dependent. They are early events, progressively achieved in the course of the apoptotic process involving lateral membrane movements of molecules. PMID- 12589728 TI - Cell-matrix interactions of in vitro human skin fibroblasts upon addition of hyaluronan. AB - Normal human skin fibroblasts were grown in a three-dimensional collagen gel or in monolayer in the presence or absence of high molecular weight hyaluronan (HA) to assess the influence of extracellular HA on cell-matrix interactions. HA incorporated into the collagen gel or added to the culture medium did not modify lattice retraction with time. The effect was independent from HA molecular weight (from 7.5 x 10(5) to 2.7 x 10(6) Da) and concentration (from 0.1 up to 1 mg/ml). HA did not affect shape and distribution of fibroblasts within the gel, whereas it induced the actin filaments to organise into thicker cables running underneath the plasma membrane. The same phenomenon was observed in fibroblasts grown in monolayer. By contrast, vimentin cytoskeleton and cell-substrate focal adhesions were not modified by exogenous HA. The number of fibroblasts attached to HA coated dishes was always significantly lower compared to plastic and to collagen type I-coated plates. By contrast, adhesion was not affected by soluble HA added to the medium nor by anti-CD44 and anti-RHAMM-IHABP polyclonals. After 24-h seeding on collagen type I or on plastic, cells were large and spread. Conversely, cells adherent to HA-coated surfaces were long, thin and aligned into rows; alcian blue showed that cells were attached to the plastic in between HA bundles. Therefore, normal human skin fibroblasts exhibit very scarce, if any, adhesion to matrix HA, either soluble or immobilised. Moreover, even at high concentration, HA molecules do not exert any visco-mechanical effect on lattice retraction and do not interfere with fibroblast-collagen interactions nor with focal adhesion contacts of fibroblasts with the substrate. This is probably relevant in organogenesis and wound repair. By contrast, HA greatly modifies the organisation of the actin cytoskeleton, suggesting that CD44-mediated signal transduction by HA may affect cell locomotion and orientation, as indicated by the fusiform shape of fibroblasts grown in the presence of immobilised HA. A role of HA in cell orientation could be relevant for the deposition of collagen fibrils in regeneration and tissue remodelling. PMID- 12589729 TI - Altered morphology in cultured rat intestinal epithelial IEC-6 cells is associated with alkaline phosphatase expression. AB - Non-transformed, rat intestinal epithelial cells (IEC-6), and human intestinal colonic carcinoma cells (CACO-2) have both been used to study processes of epithelial cell differentiation. However, only CACO-2 cells have been described as spontaneously expressing phenotypic changes of differentiation in culture. We report here that when IEC-6 cells are grown in post-confluent culture, they develop structural changes similar to those seen in cells induced to differentiate by culture on Englebreth-Holm-Swarm (EHS) extracellular matrix proteins. Correlated with this morphological change is loss of nuclear localization of c-myc protein and development of cell surface alkaline phosphatase (ALP) enzymatic activity. Messenger RNAs for liver and intestinal isoforms of ALP were expressed in both pre- and post-confluent cells. Inhibition of ALP activity in post-confluent cells by levamisole indicated the expressed ALP activity to be of the liver isoform. We suggest the expression of ALP activity, which occurs concomitantly with morphological alterations in post-confluent IEC-6 cells, represents increased expression and localization to the cell surface of the liver isoform of ALP. Cultured IEC-6 cells may provide a non-transformed, in vitro alternative to CACO-2 cells for study of epithelial cell differentiation. PMID- 12589730 TI - Macrophage lipid body induction by Chagas disease in vivo: putative intracellular domains for eicosanoid formation during infection. AB - Lipid bodies (LB), lipid-rich inclusions abundantly present in cells engaged in inflammation, are specialized intracellular domains involved in generating inflammatory mediators, the eicosanoids. Since the acute Trypanosoma cruzi infection triggers a potent inflammatory reaction characterized by a great increase of peripheral blood monocyte (PBM) and macrophage numbers, we investigated the LB occurrence in these cells. The experimental rat infection by T. cruzi (Y strain) induced significant increase of the LB numbers in peritoneal macrophages at day 6 and 12, accompanied by significant enhancement of Prostaglandin E(2) (PGE(2)) production, as measured by EIA. At day 12, ultrastructural analysis of the heart, a target organ of the disease, showed numerous macrophages with LB prominently increased in number (mean of 8.3 per section view, range of 1-25) compared to controls (mean of 2.6 per section view, range of 0-3) and size. PBM from all groups rarely showed LB. Our results demonstrate, for the first time, that T. cruzi infection in rats elicits important LB formation in inflammatory macrophages but not in PBM. The increase in LB numbers during infection positively correlates with increased generation of PGE(2), suggesting that LB may have a role in the heightened eicosanoid production observed during T. cruzi infection. PMID- 12589731 TI - In vitro chondrogenesis of human bone marrow-derived mesenchymal progenitor cells in monolayer culture: activation by transfection with TGF-beta2. AB - Bone marrow-derived mesenchymal progenitor cells are capable of chondrogenesis, making them a possible source of cells for cartilage tissue engineering. Because of this, we studied the effect of human transforming growth factor beta2 (TGF beta2) on mesenchymal progenitor cell chondrogenesis in monolayer culture using gene transfection technology. A recombinant pcDNA3.1(+)/TGF-beta2 construct containing a full-length TGF-beta2 from a human placental cDNA library was created through gene cloning and DNA recombination. The construct was then lipofected into mesenchymal progenitor cells isolated from human bone marrow. RT PCR, Western blotting, and immunohistochemistry analyses were performed to identify the expression of TGF-beta2 and cartilage-associated genes and proteins. The results showed that TGF-beta2 was expressed throughout the culture period. The transfected progenitor cells expressed and produced collagen type II and aggrecan 48 h after transfection, and the expression and synthesis were upregulated after 4 weeks. In contrast, the tested genes and proteins were not detected in non-transfected cells. This shows that transfection of pcDNA3.1(+)/TGF-beta2 into mesenchymal progenitor cells is able to provide transient and persistent expression of cartilage-specific genes and proteins, and suggests that the differentiation of human marrow-derived mesenchymal progenitor cells into chondrocytes in monolayer culture is feasible and may be induced by TGF-beta2. PMID- 12589732 TI - The role of stem cells in the evolution of longevity and its application to tissue therapy. AB - The role of stem cells in a multi-tiered organization of tissue structure extends the potential of longevity in multicelled organisms. This role can be responsible for the mechanism of evolution of the stem cells themselves. The principles involved in this mechanism can aid research for the development and management of successful tissue therapies. PMID- 12589733 TI - Mycoplasma bovis: disease, diagnosis, and control. AB - Mycoplasma bovis is a major, but often overlooked, pathogen causing respiratory disease, mastitis, and arthritis in cattle. It is found worldwide and has spread into new areas, including Ireland and parts of South America, in the last decade. In Europe, it is responsible for at least a quarter to a third of all calf pneumonia although this may be an underestimate as few laboratories regularly monitor for mycoplasmas. Like all mollicutes, M. bovis is inherently refractory to certain groups of antibiotics because it does not possess a cell wall; furthermore evidence is accumulating that strains of M. bovis are becoming resistant to antibiotics, including tetracycline, tilmicosin and spectinomycin, traditionally used for their control. No vaccines are presently available for the control of M. bovis infections. PMID- 12589734 TI - Identification and quantification of amines in the equine caecum. AB - Acute laminitis has been associated with the release of compounds, as yet unidentified, produced by hindgut fermentation which affect blood flow to the digit. The objectives of this study were to identify amine compounds in equine caecal and colonic contents, some of which are known to have vasoactive properties. In addition, the concentrations of amines in caecal contents of horses fed either grass or hay diets were compared. Fifteen amines were identified in equine hindgut contents in concentrations greater than 1 microM. The caecal concentrations of phenylethylamine, isoamylamine, cadaverine, diaminoheptane and spermidine were significantly higher in horses on spring/summer grass compared with those on winter grass or hay. These data show that many amines are present in the equine hindgut, some of which may have the potential to cause peripheral vasoconstriction if released into the circulation from the gastrointestinal tract. PMID- 12589735 TI - Effect of handling-induced stress on the results of spectral Doppler echocardiography in falcons. AB - In order to evaluate the effects of habituation to handling on the Doppler derived velocity of diastolic ventricular inflow and systolic aortic outflow, falcons that were accustomed to regular handling were compared with falcons that had been handled very little. The mean heart rate of the frequently handled birds was 111 beats per minute less than the non-habituated birds. All the mean blood flow velocities recorded were much higher in the non-habituated birds than those accustomed to handling. Taking into account the difficulty of objectively quantifying the level of stress suffered by any individual bird, the question is, if meaningful clinical or reference values for Doppler-derived blood flow velocities may be obtained in unsedated or non-anaesthetized birds? PMID- 12589736 TI - A comparison of the rate of recovery of 13CO2 in exhaled breath with 2H in body water following ingestion of [2H/13C]octanoic acid in a dog. AB - The objective of this study was to assess post-gastric processing of octanoic acid (OA) by comparing the rate of recovery of 13C in breath with the rate of recovery of 2H in saliva following ingestion of [2H/13C]OA. A test meal with 100mg [13C]OA and 750 mg [2H]OA was ingested on two separate occasions by one dog. Exhaled breath and saliva samples were collected at set time points for 6h following ingestion of the test meal. Two indices were computed, half recovery time (t(1/2)) and the time of peak excretion (t(max)). Recovery of 2H in saliva was quicker than recovery of 13C in breath as determined by the difference in the indices for each isotope; 1.33 and 1.59 h for t(1/2) and t(max), respectively. These findings suggest that the post-gastric processing of OA imposes a delay on the recovery of 13C in breath in the dog, as occurs in man and in the horse. PMID- 12589737 TI - Influence of different tumour types on natural cytotoxicity (NK cell activity) and mitogen-induced lymphocyte proliferation in isolated blood lymphocytes from 110 dogs with tumours. AB - The cell-mediated immune response of blood lymphocytes from 110 untreated dogs with different tumours was evaluated. The influence of different tumour types on the cellular immune system was examined by assessing the percentage of isolated large granular lymphocytes (LGL), in vitro natural cytotoxicity and mitogen induced lymphocyte proliferation. Although the overall natural cytotoxicity of dogs with different tumours was decreased, the overall difference from control values was not statistically significant. However, mitogen-induced lymphocyte proliferation was significantly depressed in dogs with tumours in comparison with the controls. Dogs with mammary carcinomas showed significantly lower natural cytotoxicity than controls and dogs with myeloid neoplasms showed significantly lower mitogen-induced lymphocyte proliferation. Abnormalities exist not only in natural cytotoxicity but also in mitogen-induced lymphocyte proliferation. For the dog, this is the first study to assess the influence of different tumours using a combined evaluation of natural cytotoxicity and mitogen-induced lymphocyte proliferation in such a large number of animals. PMID- 12589738 TI - Assessing the severity of canine pancreatitis. AB - The objective of this study was to determine whether laboratory testing currently available is able to provide prognostic information in canine pancreatitis. A prospective study of dogs with naturally occurring pancreatitis was undertaken. Twenty-two cases with histologically confirmed pancreatic inflammation were included in the study. Each dog had routine haematology parameters, serum biochemistry (including lipase and amylase), serum trypsin-like immunoreactivity and trypsinogen activation peptides (TAP) in urine and plasma measured. Twelve of the dogs were classified as having severe disease. These dogs had statistically significant increases in urinary TAP-creatinine ratio (UTCR) measurement, serum lipase, serum phosphate and serum creatinine concentrations. Additionally dogs with severe pancreatitis had significantly decreased urine specific gravity levels. The most sensitive and specific test to assess the severity of pancreatitis was the measurement of UTCR. PMID- 12589739 TI - Effect of dietary moisture and sodium content on urine composition and calcium oxalate relative supersaturation in healthy miniature schnauzers and labrador retrievers. AB - The aim of this series of studies was to evaluate two possible feeding strategies as methods for reducing the risk of calcium oxalate (CaOx) formation in two breeds of healthy dog. The studies compared the effect of dietary moisture (Study 1) and dietary sodium (Na), (Study 2) on urine composition of labrador retrievers (LR) and miniature schnauzers (MS). A nutritionally complete dry dog food was fed to 16 dogs (eight LR, eight MS; Study 1) and 15 dogs (seven LR, eight MS; Study 2) for 24 days (Study 1), or 36 days (Study 2). The dogs were fed the diet alone (7% moisture, 0.06 g Na/100 kcal), or supplemented with deionised water to 73% moisture (Study 1), or dietary Na, to deliver 0.20 or 0.30 g Na per 100 kcal (Study 2). Urine pH, volume, specific gravity, and concentrations of 12 analytes were measured for each dog. Urinary relative supersaturations (RSS) with CaOx were calculated from these values. The effects of supplemental Na or water were established using t tests (Study 1) or analysis of variance, and multiple range tests (least significant difference) (Study 2); P<0.05 was considered significant. Increasing dietary moisture significantly increased total moisture intake (P=0.001), and reduced urine specific gravity (P=0.003), urinary oxalate concentration (P=0.04), and CaOx relative supersaturation (P=0.04) in the MS. Urinary parameters remained unchanged in the LR, indicating that feeding a high moisture diet may reduce the risk of CaOx formation in high-risk breeds. Increasing dietary Na led to production of urine with a significantly lower CaOx RSS in both breeds, indicating that sodium supplementation to dry diet formats may reduce the risk of CaOx formation. These feeding strategies should be considered when evaluating methods for preventing CaOx formation within high-risk groups. PMID- 12589740 TI - A morphological study of simple testicular cysts in the ostrich (Struthio camelus). AB - Simple testicular cysts are rare in mammals and of unknown aetiology, but dietary conditions have been implicated in this phenomenon in poultry. This study characterises the structural features of simple intratesticular cysts in the ostrich. Seven testes from a total of 15 sexually mature ostriches slaughtered during the breeding season were used. The testes contained multifocal, fluid filled, unilocular cysts which were lined by a simple squamous or low cuboidal epithelium and surrounded by a thick layer of fibrous connective tissue. The cysts developed within seminiferous tubules and displayed morphological features similar to those reported in man and domestic poultry. The testis parenchyma revealed several foci composed of intermingled normal seminiferous tubules and variably sized intratesticular cysts. The atrophic tubules lay within a mass of hyperplastic, fibroblastic intertubular connective tissue in the proximity of large cysts and their formation appeared to result from hydrostatic pressure exerted by cysts. Morphological evidence supports a continuous process of cyst formation in the affected testis and a concomitant progressive loss of atrophic seminiferous tubules. A pathogenetic scenario of cyst formation and the effect of simple cysts on testicular histology has been proposed. Although the course of this phenomenon remains unknown, its impact on the fertility of this economically important bird deserves closer scrutiny. PMID- 12589741 TI - Diarrhoea in the growing pig - a comparison of clinical, morphological and microbial findings between animals from good and poor performance herds. AB - Diarrhoea among growing pigs (8-13 weeks old) is a significant problem in many herds. Nine herds with poor performance and diarrhoea among growing pigs were selected on the basis of their piglet mean age at a body weight of 25 kg, compared to the overall mean age in Swedish herds. In addition, four herds with good average performance and no problems with diarrhoea were selected. Pigs were necropsied and samples for histology and microbiology were collected. Based on the necropsy findings, the pigs from the good performing herds were all judged to be healthy. The presence of Brachyspira pilosicoli and Lawsonia intracellularis was significantly correlated to poor performing herds and the results indicate that these microbes are main pathogens involved in enteric diseases among Swedish grower pigs. In addition, concomitant infections with other presumptive pathogens were commonly found. PMID- 12589742 TI - Efficacy of danofloxacin 18% injectable solution in the treatment of Escherichia coli diarrhoea in young calves in Europe. AB - The efficacy of danofloxacin 18% against naturally occurring Escherichia coli diarrhoea was investigated in calves at seven European sites. Treatment commenced on day 0, with either a single subcutaneous injection of danofloxacin 18% (n=267) at 6 mg/kg repeated on day 2 if required, or reference product containing baquiloprim/sulphadimidine (n=37) or gentamicin (n=98) administered as recommended. E. coli was isolated from 90% to 100% of calves pre-treatment, and the prevalence of serotypes K99 and F41 was 8-46% and 46-92%, respectively. In both treatments, the majority of calves (93.2-93.9%) showed clinical improvement and completed the studies. There were significant reductions for both treatments, in severity of clinical signs on days 4 and 10 compared to day 0 (P<0.0001), and between days 4 and 10 (P<0.05), but no significant differences between treatments (P>0.05). Danofloxacin 18% was clinically safe, and as effective as the reference products in the treatment of E. coli diarrhoea in calves. PMID- 12589743 TI - Absence of urinary tract infection in dogs with experimentally induced hyperadrenocorticism. AB - Objectives of this study were to determine occurrence of urinary tract infection and describe results of urine analysis and urine culture in dogs with experimentally induced hyperadrenocorticism. Dogs were randomly assigned to receive either hydrocortisone (nine dogs) or placebo (eight dogs) for 49 consecutive days. Before and on day 49 of treatment, evaluation of dogs included physical examination, abdominal ultrasound, urine culture, urinalysis, adrenal function testing, and measurement of urine protein and creatinine and activity of serum alkaline phosphatase. All dogs in the experimental group had clinical and laboratory findings of hyperadrenocorticism. Urine specific gravity was significantly decreased and urine protein-to-creatinine ratio was significantly increased in dogs with hyperadrenocorticism. Urinary tract infection did not occur in any dogs. We conclude that administration of hydrocortisone created a model of hyperadrenocorticism; however, urinary tract infection did not occur. Additional evaluation is needed to determine association between urinary tract infection and hyperadrenocorticism. PMID- 12589744 TI - Decrease in malate dehydrogenase activities in peripheral leucocytes of type 1 diabetic dogs. AB - The activities of the enzymes involved in the malate-aspartate shuttle were measured in peripheral leucocytes of dogs with type 1 diabetes mellitus. In the diabetic dogs, fasting plasma glucose concentrations were twofold greater than control levels despite insulin injections and the activities of malate dehydrogenase (MDH), which plays a crucial role in the malate-aspartate shuttle, were decreased remarkably. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity (M/L ratio) in leucocytes of the diabetic dogs was significantly lower than that of normal control dogs. The decrease of the M/L ratio appeared to reflect depression of energy metabolism in leucocytes of the diabetic dogs. The M/L ratio may be a useful parameter to evaluate metabolic conditions in diabetic dogs. PMID- 12589745 TI - Immunogenicity of bovine parainfluenza type 3 virus proteins encapsulated in nanoparticle vaccines, following intranasal administration to mice. AB - Stimulation of long lasting, protective immunity to respiratory viruses is often difficult to achieve with conventional respiratory vaccines. Polymeric nanoparticles, incorporating viral proteins have been shown to offer sustained release of antigen, with consequent prolongued stimulation of the respiratory immune system. In this paper the efficacy of two nanoparticle vaccines (poly lactide-co-glycolide, PLGA; polymethylmethacrylate, PMMA), incorporating proteins of bovine parainfluenza type 3 virus (BPI-3) was investigated. As a preliminary to experiments in calves, it was considered essential to demonstrate immunogenicity of the experimental vaccine in mice. Mice immunised with PLGA nanoparticles, containing BPI-3 proteins, developed higher levels of virus specific antibody than mice immunised with the PMMA vaccine or with soluble viral proteins alone. Immunoblotting using serum from the vaccinated mice, demonstrated strong reactions against the major BPI-3 proteins. PMID- 12589746 TI - Transient kinetic analysis of ATP-induced allosteric transitions in the eukaryotic chaperonin containing TCP-1. AB - The chaperonin CCT (chaperonin containing t-complex polypeptide 1 (TCP-1)) from bovine testis was mixed rapidly with different concentrations of ATP and the time resolved change in fluorescence emission, upon excitation at 280 nm, was followed. Two kinetic phases were observed and assigned by (i) analyzing the dependence of the corresponding observed rate constants on ATP concentration; and (ii) by carrying out mixing experiments also with ADP, ATPgammaS and ATP without K(+). The values of the observed rate constants corresponding to both phases are found to be dependent on ATP concentration. The observed rate constant corresponding to the fast phase displays a bi-sigmoidal dependence on ATP concentration with Hill coefficients that are similar to those determined in steady-state ATPase experiments. This phase most likely reflects ATP binding induced conformational changes. The rate constant of the conformational change in the presence of excess ATP is about 17s(-1) (at 25 degrees C) and is tenfold slower than the corresponding rate constant of GroEL. The observed rate constant corresponding to the second slower phase displays a hyperbolic dependence on ATP concentration. This phase is not observed in mixing experiments of CCT with ADP, ATPgammaS or ATP without K(+) and it, therefore, reflects a conformational change associated with ATP hydrolysis. Taken together, our results indicate that the kinetic mechanism of the allosteric transitions of CCT differs considerably from that of GroEL. PMID- 12589747 TI - Solution structure of the chick TGFbeta type II receptor ligand-binding domain. AB - The transforming growth factor beta (TGFbeta) signaling pathway influences cell proliferation, immune responses, and extracellular matrix reorganization throughout the vertebrate life cycle. The signaling cascade is initiated by ligand-binding to its cognate type II receptor. Here, we present the structure of the chick type II TGFbeta receptor determined by solution NMR methods. Distance and angular constraints were derived from 15N and 13C edited NMR experiments. Torsion angle dynamics was used throughout the structure calculations and refinement. The 20 final structures were energy minimized using the generalized Born solvent model. For these 20 structures, the average backbone root-mean square distance from the average structure is below 0.6A. The overall fold of this 109-residue domain is conserved within the superfamily of these receptors. Chick receptors fully recognize and respond to human TGFbeta ligands despite only 60% identity at the sequence level. Comparison with the human TGFbeta receptor determined by X-ray crystallography reveals different conformations in several regions. Sequence divergence and crystal packing interactions under low pH conditions are likely causes. This solution structure identifies regions were structural changes, however subtle, may occur upon ligand-binding. We also identified two very well conserved molecular surfaces. One was found to bind ligand in the crystallized human TGFbeta3:TGFbeta type II receptor complex. The other, newly identified area can be the interaction site with type I and/or type III receptors of the TGFbeta signaling complex. PMID- 12589749 TI - Voltage-gated K+ channel from mammalian brain: 3D structure at 18A of the complete (alpha)4(beta)4 complex. AB - Voltage-sensitive K(+) channels (Kv) serve numerous important roles, e.g. in the control of neuron excitability and the patterns of synaptic activity. Here, we use electron microscopy (EM) and single particle analysis to obtain the first, complete structure of Kv1 channels, purified from rat brain, which contain four transmembrane channel-forming alpha-subunits and four cytoplasmically-associated beta-subunits. The 18A resolution structure reveals an asymmetric, dumb-bell shaped complex with 4-fold symmetry, a length of 140A and variable width. By fitting published X-ray data for recombinant components to our EM map, the modulatory (beta)(4) was assigned to the innermost 105A end, the N-terminal (T1)(4) domain of the alpha-subunit to the central 50A moiety and the pore containing portion to the 125A membrane part. At this resolution, the selectivity filter could not be localised. Direct contact of the membrane component with the central (T1)(4) domain occurs only via peripheral connectors, permitting communication between the channel and beta-subunits for coupling of responses to changes in excitability and metabolic status of neurons. PMID- 12589748 TI - Inherent protein structural flexibility at the RNA-binding interface of L30e. AB - The Saccharomyces cerevisiae ribosomal protein L30 autoregulates its own expression by binding to a purine-rich internal loop in its pre-mRNA and mRNA. NMR studies of L30 and its RNA complex showed that both the internal loop of the RNA as well as a region of the protein become substantially more ordered upon binding. A crystal structure of a maltose binding protein (MBP)-L30 fusion protein with two copies in the asymmetric unit has been determined. The flexible RNA-binding region in the L30 copies has two distinct conformations, one resembles the RNA bound form solved by NMR and the other is unique. Structure prediction algorithms also had difficulty accurately predicting this region, which is consistent with conformational flexibility seen in the NMR and X-ray crystallography studies. Inherent conformational flexibility may be a hallmark of regions involved in intermolecular interactions. PMID- 12589750 TI - Mycobacterium tuberculosis Hsp16.3 nonamers are assembled and re-assembled via trimer and hexamer intermediates. AB - Hsp16.3, a small heat shock protein from Mycobacterium tuberculosis proposed to form specific trimer-of-trimers structures, acts as a molecular chaperone in vitro. The assembly and re-assembly mechanisms of this oligomeric protein were studied and compared using in vitro transcription/translation and denaturization/renaturization systems. Analysis using a combination of non denaturing pore gradient polyacrylamide gel electrophoresis, chemical cross linking, and size-exclusion chromatography demonstrate that the predominant form of Hsp16.3 produced in the in vitro transcription/translation system is the trimer, which can be further assembled into a nonameric structure via a hexamer intermediate in the presence of purified exogenous Hsp16.3 proteins. Meanwhile, an "inert" Hsp16.3 dimer, which does not seem to participate in nonamer assembly but may be involved in forming other forms of Hsp16.3, was also detected in the in vitro expression system. On the other hand, our current data clearly show that the re-assembly of Hsp16.3 nonamers also occurs via a very similar mechanism, with the formation of trimers and hexamers. The presence of high levels of macromolecular crowding protein agent in the in vitro expression system promoted the formation of the nonamers to a very limited degree, indicating that the assembly of proteins like Hsp16.3 may depend mainly on its own concentration instead of those of the macromolecules in the environment. PMID- 12589751 TI - Substrate complexes of hepatitis C virus RNA polymerase (HC-J4): structural evidence for nucleotide import and de-novo initiation. AB - Several crystal structures of the hepatitis C virus NS5B protein (genotype-1b, strain J4) complexed with metal ions, single-stranded RNA or nucleoside triphosphates have been determined. These complexes illustrate how conserved amino acid side-chains, together with essential structural features within the active site, control nucleotide binding and likely mediate de-novo initiation. The incoming nucleotide interacts with several basic residues from an extension on the NS5B fingers domain, a beta-hairpin from the NS5B thumb domain and the C terminal arm. The modular, bi-partite fingers domain carries a long binding groove which guides the template towards the catalytic site. The apo-polymerase structure provides unprecedented insights into potential non-nucleoside inhibitor binding sites located between palm and thumb near motif E, which is unique to RNA polymerases and reverse transcriptases. PMID- 12589752 TI - Refined solution structure of the iron-responsive element RNA using residual dipolar couplings. AB - The iron-responsive element (IRE) is a 30nt RNA motif located in the non-coding regions of mRNAs of proteins involved in iron regulation. In humans, the IRE plays a direct role in the control of iron levels by post-transcriptional regulation of the ferritin and transferrin receptor proteins through highly specific recognition by IRE-binding proteins. The IRE fold is representative of many RNA motifs that contain helical domains separated by a bulge or internal loop. The global structures of such extended multi-domain RNAs are not well defined by conventional NMR-distance and torsion angle structural restraints. Residual dipolar couplings (RDCs) are employed here to better define the global structure of the IRE RNA in solution. RDCs contain valuable long-range structural information that compliments the short-range structural data derived from standard NOE-distance and torsion angle restraints. Several approaches for estimating alignment tensor parameters and incorporating RDCs into RNA structure determinations are compared. Both the local and global structure of the IRE are improved significantly by refinement with RDCs. These RDC refinements provide insight on the conformational dynamics of the IRE. These studies highlight some issues that need to be addressed when incorporating RDCs in solution structure determinations of nucleic acids. The approach used here should prove valuable for structure determinations of various multi-domain systems. PMID- 12589753 TI - The metal-independent type IIs restriction enzyme BfiI is a dimer that binds two DNA sites but has only one catalytic centre. AB - BfiI is a novel type IIs restriction endonuclease that, unlike all other restriction enzymes characterised to date, cleaves DNA in the absence of Mg(2+). The amino acid sequence of the N-terminal part of BfiI has some similarities to Nuc of Salmonella typhimurium, an EDTA-resistant nuclease akin to phospholipase D. The dimeric form of Nuc contains a single active site composed of residues from both subunits. To examine the roles of the amino acid residues of BfiI that align with the catalytic residues in Nuc, a set of alanine replacement mutants was generated by site-directed mutagenesis. The mutationally altered forms of BfiI were all catalytically inactive but were still able to bind DNA specifically. The active site of BfiI is thus likely to be similar to that of Nuc. BfiI was also found by gel-filtration to be a dimer in solution. Both gel shift and pull-down assays indicated that the dimeric form of BfiI binds two copies of its recognition sequence. In reactions on plasmids with either one or two copies of its recognition sequence, BfiI cleaved the DNA with two sites more rapidly than that with one site. Yet, when bound to two copies of its recognition sequence, the BfiI dimer cleaved only one phosphodiester bond at a time. The dimer thus seems to contain two DNA-binding domains but only one active site. PMID- 12589754 TI - Annotating nucleic acid-binding function based on protein structure. AB - Many of the targets of structural genomics will be proteins with little or no structural similarity to those currently in the database. Therefore, novel function prediction methods that do not rely on sequence or fold similarity to other known proteins are needed. We present an automated approach to predict nucleic-acid-binding (NA-binding) proteins, specifically DNA-binding proteins. The method is based on characterizing the structural and sequence properties of large, positively charged electrostatic patches on DNA-binding protein surfaces, which typically coincide with the DNA-binding-sites. Using an ensemble of features extracted from these electrostatic patches, we predict DNA-binding proteins with high accuracy. We show that our method does not rely on sequence or structure homology and is capable of predicting proteins of novel-binding motifs and protein structures solved in an unbound state. Our method can also distinguish NA-binding proteins from other proteins that have similar, large positive electrostatic patches on their surfaces, but that do not bind nucleic acids. PMID- 12589755 TI - The fission yeast spSet1p is a histone H3-K4 methyltransferase that functions in telomere maintenance and DNA repair in an ATM kinase Rad3-dependent pathway. AB - We have characterized spSet1p, the Schizosaccharomyces pombe ortholog of the budding yeast histone H3 methyltransferase Set1p. SpSet1p catalyzes methylation of H3 at K4, in vivo and in vitro. Deleting spset1 partially affects telomeric and centromeric silencing. Strikingly, lack of spSet1p causes elongation of telomeres in wild-type cells and in most DNA damage checkpoint rad mutant cells, but not in cells lacking the ATM kinase Rad3 or its associated protein Rad26. Interestingly, spset1 deletion specifically causes a reduction in sensitivity to ultraviolet radiation of the PCNA-like checkpoint mutants hus1 and rad1, but not of cells devoid of Rad3. This partial suppression was not due to restoration of checkpoint function or to transcriptional induction of DNA repair genes. Moreover, spset1 allows recovery specifically of the crb2 checkpoint mutant upon treatment with the replication inhibitor hydroxyurea but not upon UV irradiation. Nevertheless, the pathway induced in spset1 cells cannot substitute for the Mus81/Rqh1 DNA damage tolerance pathway. Our results suggest that SpSet1p and the ATM kinase Rad3 function in a common genetic pathway linking chromatin to telomere length regulation and DNA repair. PMID- 12589756 TI - Unpaired structures in SCA10 (ATTCT)n.(AGAAT)n repeats. AB - A number of human hereditary diseases have been associated with the instability of DNA repeats in the genome. Recently, spinocerebellar ataxia type 10 has been associated with expansion of the pentanucleotide repeat (ATTCT)(n).(AGAAT)(n) from a normal range of ten to 22 to as many as 4500 copies. The structural properties of this repeat cloned in circular plasmids were studied by a variety of methods. Two-dimensional gel electrophoresis and atomic force microscopy detected local DNA unpairing in supercoiled plasmids. Chemical probing analysis indicated that, at moderate superhelical densities, the (ATTCT)(n).(AGAAT)(n) repeat forms an unpaired region, which further extends into adjacent A+T-rich flanking sequences at higher superhelical densities. The superhelical energy required to initiate duplex unpairing is essentially length-independent from eight to 46 repeats. In plasmids containing five repeats, minimal unpairing of (ATTCT)(5).(AGAAT)(5) occurred while 2D gel analysis and chemical probing indicate greater unpairing in A+T-rich sequences in other regions of the plasmid. The observed experimental results are consistent with a statistical mechanical, computational analysis of these supercoiled plasmids. For plasmids containing 29 repeats, which is just above the normal human size range, flanked by an A+T-rich sequence, atomic force microscopy detected the formation of a locally condensed structure at high superhelical densities. However, even at high superhelical densities, DNA strands within the presumably compact A+T-rich region were accessible to small chemicals and oligonucleotide hybridization. Thus, DNA strands in this "collapsed structure" remain unpaired and accessible for interaction with other molecules. The unpaired DNA structure functioned as an aberrant replication origin, in that it supported complete plasmid replication in a HeLa cell extract. A model is proposed in which unscheduled or aberrant DNA replication is a critical step in the expansion mutation. PMID- 12589757 TI - Pausing of DNA polymerases on duplex DNA templates due to ligand binding in vitro. AB - Using the recently developed peptide nucleic acid (PNA)-assisted assay, which makes it possible to extend a primer on duplex DNA, we study the sequence specific inhibition of the DNA polymerase movement along double-stranded DNA templates imposed by DNA-binding ligands. To this end, a plasmid vector has been prepared featuring the polylinker with two flanking priming sites to bi directionally initiate the primer-extension reactions towards each other. Within this plasmid, we have cloned a set of random DNA sequences and analyzed the products of these reactions with several phage and bacterial DNA polymerases capable of strand-displacement synthesis. Two of them, o29 and modified T7 (Sequenase 2.0) enzymes, were found to be most potent for primer extension in the presence of DNA-binding ligands. We used these enzymes for a detailed study of ligand-induced pausing effects with four ligands differing in modes of binding to the DNA double-helix. GC-specific intercalator actinomycin D and three minor groove-binders, chromomycin A(3) (GC-specific), distamycin A and netropsin (both AT-specific), have been chosen. In the presence of each ligand both selected DNA polymerases experienced multiple clear-cut pauses. Each ligand yielded its own characteristic pausing pattern for a particular DNA sequence. The majority of pausing sites could be located with a single-nucleotide resolution and corresponded to the preferred binding sites known from the literature for the ligands under study. Besides, DNA polymerases stalled exactly at the positions occupied by PNA oligomers that were employed to initiate the primer extension. These findings provide an important insight into the DNA polymerase performance. In addition, the high-resolution ligand-induced pausing patterns we obtained for the first time for DNA polymerase elongation on duplex DNA may become a valuable addition to the existing arsenal of methods used to monitor duplex DNA interactions with various DNA-binding ligands, including drugs. PMID- 12589758 TI - LINE-1 preTa elements in the human genome. AB - The preTa subfamily of long interspersed elements (LINEs) is characterized by a three base-pair "ACG" sequence in the 3' untranslated region, contains approximately 400 members in the human genome, and has low level of nucleotide divergence with an estimated average age of 2.34 million years old suggesting that expansion of the L1 preTa subfamily occurred just after the divergence of humans and African apes. We have identified 362 preTa L1 elements from the draft human genomic sequence, investigated the genomic characteristics of preTa L1 insertions, and screened individual elements across diverse human populations and various non-human primate species using polymerase chain reaction (PCR) assays to determine the phylogenetic origin and levels of human genomic diversity associated with the L1 elements. All of the preTa L1 elements analyzed by PCR were absent from the orthologous positions in non-human primate genomes with 33 (14%) of the L1 elements being polymorphic with respect to insertion presence or absence in the human genome. The newly identified L1 insertion polymorphisms will prove useful as identical by descent genetic markers for the study of human population genetics. We provide evidence that preTa L1 elements show an integration site preference for genomic regions with low GC content. Computational analysis of the preTa L1 elements revealed that 29% of the elements amenable to complete sequence analysis have apparently escaped 5' truncation and are essentially full-length (approximately 6kb). In all, 29 have two intact open reading frames and may be capable of retrotransposition. PMID- 12589759 TI - Full-length rat amylin forms fibrils following substitution of single residues from human amylin. AB - Pancreatic amyloid deposits, composed of the 37 amino acid residue peptide amylin, represent an integral part of type 2 diabetes mellitus pathology. Human amylin (hA) forms fibrils in vitro and is toxic to cultured pancreatic islet beta cells. In contrast, rat amylin (rA) which differs from hA by only six amino acid residues in the central region of the peptide, residues 18-29, does not form fibrils and is not cytotoxic. To elucidate the role of individual residues in fibril formation, we have generated a series of full-length rA variants and examined their ability to form fibrils in vitro. Single-residue substitutions with amino acids from corresponding positions of the hA sequence, i.e. R18H, L23F, or V26I, were sufficient to render rA competent for fibril formation albeit at a small yield. Combining two or three of these substitutions generally increased the ability to produce fibrils. Variant rA fibril morphologies were examined by negative stain electron microscopy and found to be similar to those generated by hA itself. Bulk assays, i.e. involving thioflavin-T fluorescence and sedimentation, showed that the amount of fibril formation was relatively small for these rA variants when compared to hA under the same conditions. Fibril growth was demonstrated by time-lapse atomic force microscopy, and MALDI-TOF mass spectrometry was used to verify that fibrils consisted of full-length peptide. Our observations confirm previous reports that the three proline residues play a dominant negative role in fibril formation. However, their presence is not sufficient to completely abolish the ability of rA to form fibrils, as each of the other three implicated residues (i.e. R18, L23 and V26) also has a dominant modulating effect. PMID- 12589760 TI - Crystal structure of MHC class II I-Ab in complex with a human CLIP peptide: prediction of an I-Ab peptide-binding motif. AB - Association between the class II major histocompatibility complex (MHC) and the class II invariant chain-associated peptide (CLIP) occurs naturally as an intermediate step in the MHC class II processing pathway. Here, we report the crystal structure of the murine class II MHC molecule I-A(b) in complex with human CLIP at 2.15A resolution. The structure of I-A(b) accounts, via the peptide binding groove's unique physicochemistry, for the distinct peptide repertoire bound by this allele. CLIP adopts a similar conformation to peptides bound by other I-A alleles, reinforcing the notion that CLIP is presented as a conventional peptide antigen. When compared to the related HLA-DR3/CLIP complex structure, the CLIP peptide displays a slightly different conformation and distinct interaction pattern with residues in I-A(b). In addition, after examining the published sequences of peptides presented by I-A(b), we discuss the possibility of predicting peptide alignment in the I-A(b) binding groove using a simple scoring matrix. PMID- 12589761 TI - Crystal structure of geneticin bound to a bacterial 16S ribosomal RNA A site oligonucleotide. AB - Aminoglycosides are antibacterial molecules that decrease translation accuracy by binding to the decoding aminoacyl-tRNA site (A site) on 16S ribosomal RNA. We have solved the crystal structure of an RNA fragment containing the A site bound to geneticin at 2.40A resolution. Geneticin, also known as G418, is a gentamicin related aminoglycoside: it contains three rings that are functionalized by hydroxyl, ammonium and methyl groups. The detailed comparison of the distinctive behaviour of geneticin (binding to pro- and eukaryotic A sites) with the crystallographic, biochemical and microbiological results obtained so far for aminoglycoside-A site complexes offers new insights on the system. The two sugar rings constituting the neamine part common to most of the aminoglycosides bind to the A site, as already observed in the crystal structures solved previously with paromomycin and tobramycin. The essential hydrogen bonds involving ring I (to A1408) and ring II (to the phosphate oxygen atoms of the bulged adenine bases 1492 and 1493 and to G1494) are conserved and additional contacts are observed from ring III (to phosphate oxygen atoms of G1405 and U1406). The present work illustrates a molecular basis of the range in sensitiveness exhibited by geneticin towards common point A site mutations associated to resistance phenotypes. In addition, analysis and comparisons of the structures cast light on the role played by the conserved U1406.U1495 pair in the recognition of the A site by aminoglycosides. PMID- 12589762 TI - Solution structure of the cytotoxic RNase 4 from oocytes of bullfrog Rana catesbeiana. AB - Cytotoxic ribonucleases with antitumor activity are mainly found in the oocytes and early embryos of frogs. Native RC-RNase 4 (RNase 4), consisting of 106 residues linked with four disulfide bridges, is a cytotoxic ribonuclease isolated from oocytes of bullfrog Rana catesbeiana. RNase 4 belongs to the bovine pancreatic ribonuclease (RNase A) superfamily. Recombinant RC-RNase 4 (rRNase 4), which contains an additional Met residue and glutamine instead of pyroglutamate at the N terminus, was found to possess less catalytic and cytotoxic activities than RNase 4. Equilibrium thermal and guanidine-HCl denaturation CD measurements revealed that RNase 4 is more thermally and chemically stable than rRNase 4. However, CD and NMR data showed that there is no gross conformational change between native and recombinant RNase 4. The NMR solution structure of rRNase 4 was determined to comprise three alpha-helices and two sets of antiparallel beta sheets. Superimposition of each structure with the mean structure yielded an average root mean square deviation (RMSD) of 0.72(+/-0.14)A for the backbone atoms, and 1.42(+/-0.19)A for the heavy atoms in residues 3-105. A comparison of the 3D structure of rRNase 4 with the structurally and functionally related cytotoxic ribonuclease, onconase (ONC), showed that the two H-bonds in the N terminal pyroglutamate of ONC were not present at the corresponding glutamine residue of rRNase 4. We suggest that the loss of these two H-bonds is one of the key factors responsible for the reductions of the conformational stability, catalytic and cytotoxic activities in rRNase 4. Furthermore, the differences of side-chain conformations of subsite residues among RNase A, ONC and rRNase 4 are related to their distinct catalytic activities and base preferences. PMID- 12589763 TI - Crystal structure of HPr kinase/phosphatase from Mycoplasma pneumoniae. AB - HPr kinase/phosphatase (HPrK/P) modifies serine 46 of histidine-containing protein (HPr), the phosphorylation state of which is the control point of carbon catabolite repression in low G+C Gram-positive bacteria. To understand the structural mechanism by which HPrK/P carries out its dual, competing activities we determined the structure of full length HPrK/P from Mycoplasma pneumoniae (PD8 ID, 1KNX) to 2.5A resolution. The enzyme forms a homo-hexamer with each subunit containing two domains connected by a short loop. The C-terminal domain contains the well-described P-loop (Walker A box) ATP binding motif and takes a fold similar to phosphoenolpyruvate carboxykinase (PEPCK) from Escherichia coli as recently described in other HPrK/P structures. As expected, the C-terminal domain is very similar to the C-terminal fragment of Lactobacillus casei HPrK/P and the C-terminal domain of Staphylococcus xylosus HPrK/P; the N-terminal domain is very similar to the N-terminal domain of S.xylosus HPrK/P. Unexpectedly, the N terminal domain resembles UDP-N-acetylmuramoyl-L-alanyl-D-glutamate:meso diaminopimelate ligase (MurE), yet the function of this domain is unclear. We discuss these observations as well as the structural significance of mutations in the P-loop and HPrK/P family sequence motif. PMID- 12589764 TI - X-ray structure of a maquette scaffold. AB - Maquettes are de novo designed mimicries of nature used to test the construction and engineering criteria of oxidoreductases. One type of scaffold used in maquette construction is a four-alpha-helical bundle. The sequence of the four alpha-helix bundle maquettes follows a heptad repeat pattern typical of left handed coiled-coils. Initial designs were molten globular due partly to the minimalist approach taken by the designers. Subsequent iterative redesign generated several structured scaffolds with similar heme binding properties. Variant [I(6)F(13)](2), a structured scaffold, was partially resolved with NMR spectroscopy and found to have a set of mobile inter-helical packing interfaces. Here, the X-ray structure of a similar peptide ([I(6)F(13)M(31)](2) i.e. ([CGGG EIWKL HEEFLKK FEELLKL HEERLKKM](2))(2) which we call L31M), has been solved using MAD phasing and refined to 2.8A resolution. The structure shows that the maquette scaffold is an anti-parallel four-helix bundle with "up-up-down-down" topology. No pre-formed heme-binding pocket exists in the protein scaffold. We report unexpected inter-helical crossing angles, residue positions and translations between the helices. The crossing angles between the parallel helices are -5 degrees rather than the expected +20 degrees for typical left-handed coiled coils. Deviation of the scaffold from the design is likely due to the distribution and size of hydrophobic residues. The structure of L31M points out that four identical helices may interact differently in a bundle and heptad repeats with an alternating [HPPHHPP]/[HPPHHPH] (H: hydrophobic, P: polar) pattern are not a sufficient design criterion to generate left-hand coiled-coils. PMID- 12589765 TI - Effect of dextran on protein stability and conformation attributed to macromolecular crowding. AB - Thermally induced transition curves of hen egg-white lysozyme were measured in the presence of several concentrations of dextran at pH 2.0 by near-UV and far-UV CD. The transition curves were fitted to a two-state model by a non-linear, least squares method to obtain the transition temperature (T(m)), enthalpy change (deltaH(u)(T(m))), and free energy change (deltaG(u)(T)) of the unfolding transition. An increase in T(m) and almost constant deltaH(u)(T(m)) values were observed in the presence of added dextran at concentrations exceeding ca 100 g l( 1). In addition, dextran-induced conformational changes of fully unfolded protein were investigated by CD spectroscopy. Addition of high concentrations of dextran to solutions of acid-unfolded cytochrome c at pH 2.0 results in a shift of the CD spectrum from that characteristic of the fully unfolded polypeptide to that characteristic of the more compact, salt-induced molten globule state, a result suggesting that the molten globule-like state is stabilized relative to the fully unfolded form in crowded environments. Both observations are in qualitative accord with predictions of a previously proposed model for the effect of intermolecular excluded volume (macromolecular crowding) on protein stability and conformation. PMID- 12589766 TI - An orientation-dependent hydrogen bonding potential improves prediction of specificity and structure for proteins and protein-protein complexes. AB - Hydrogen bonding is a key contributor to the specificity of intramolecular and intermolecular interactions in biological systems. Here, we develop an orientation-dependent hydrogen bonding potential based on the geometric characteristics of hydrogen bonds in high-resolution protein crystal structures, and evaluate it using four tests related to the prediction and design of protein structures and protein-protein complexes. The new potential is superior to the widely used Coulomb model of hydrogen bonding in prediction of the sequences of proteins and protein-protein interfaces from their structures, and improves discrimination of correctly docked protein-protein complexes from large sets of alternative structures. PMID- 12589767 TI - Rapid cooperative two-state folding of a miniature alpha-beta protein and design of a thermostable variant. AB - There is a great deal of interest in developing small stably folded miniature proteins. A limited number of these molecules have been described, however they typically have not been characterized in depth. In particular, almost no detailed studies of the thermodynamics and folding kinetics of these proteins have been reported. Here we describe detailed studies of the thermodynamics and kinetics of folding of a 39 residue mixed alpha-beta protein (NTL9(1-39)) derived from the N terminal domain of the ribosomal protein L9. The protein folds cooperatively and rapidly in a two-state fashion to a native state typical of those found for normal globular proteins. At pH 5.4 in 20mM sodium acetate, 100mM NaCl the temperature of maximum stability is 6 degrees C, the t(m) is 65.3 degrees C, deltaH degrees (t(m)) is between 24.6 kcalmol(-1) and 26.3 kcalmol(-1), and deltaC(p) degrees is 0.38 kcalmol(-1)deg(-1). The thermodynamic parameters are in the range expected on the basis of per residue values determined from databases of globular proteins. H/2H exchange measurements reveal a set of amides that exchange via global unfolding, exactly as expected for a normal cooperatively folded globular protein. Kinetic measurements show that folding is two-state folding. The folding rate is 640 s(-1) and the value of deltaG degrees calculated from the folding and unfolding rates is in excellent agreement with the equilibrium value. A designed thermostable variant, generated by mutating K12 to M, was characterized and found to have a t(m) of 82 degrees C. Equilibrium and kinetic measurements demonstrate that its folding is cooperative and two-state. PMID- 12589768 TI - The thermodynamics of protein-protein recognition as characterized by a combination of volumetric and calorimetric techniques: the binding of turkey ovomucoid third domain to alpha-chymotrypsin. AB - We have used ultrasonic velocimetry, high-precision densimetry, and fluorescence spectroscopy, in conjunction with isothermal titration and differential scanning calorimetry, to characterize the binding of turkey ovomucoid third domain (OMTKY3) to alpha-chymotrypsin. We report the changes in volume and adiabatic compressibility that accompany the association of these proteins at 25 degrees C and pH 4.5. In addition, we report the changes in free energy, enthalpy, entropy, and heat capacity upon the binding of OMTKY3 to alpha-chymotrypsin over a temperature range of 20-40 degrees C. Our volume and compressibility data, in conjunction with X-ray crytsallographic data on the OMTKY3-alpha-chymotrypsin complex, suggest that 454(+/-22) water molecules are released to the bulk state upon the binding of OMTKY3 to alpha-chymotrypsin. Furthermore, these volumetric data suggest that the intrinsic compressibility of the two proteins decreases by 7%. At each temperature studied, OMTKY3 association with alpha-chymotrypsin is entropy driven with a large, unfavorable enthalpy contribution. The observed entropy of the binding reflects interplay between two very large favorable and unfavorable terms. The favorable term reflects an increase in the hydrational entropy resulting from release to the bulk of 454 water molecules. The unfavorable term is related to a decrease in the configurational entropy and, consequently, a decrease in the conformational dynamics of the two proteins. In general, we discuss the relationship between macroscopic and microscopic properties, in particular, identifying and quantifying the role of hydration in determining the thermodynamics of protein recognition as reflected in volumetric and calorimetric parameters. PMID- 12589769 TI - Automatic methods for predicting functionally important residues. AB - Sequence analysis is often the first guide for the prediction of residues in a protein family that may have functional significance. A few methods have been proposed which use the division of protein families into subfamilies in the search for those positions that could have some functional significance for the whole family, but at the same time which exhibit the specificity of each subfamily ("Tree-determinant residues"). However, there are still many unsolved questions like the best division of a protein family into subfamilies, or the accurate detection of sequence variation patterns characteristic of different subfamilies. Here we present a systematic study in a significant number of protein families, testing the statistical meaning of the Tree-determinant residues predicted by three different methods that represent the range of available approaches. The first method takes as a starting point a phylogenetic representation of a protein family and, following the principle of Relative Entropy from Information Theory, automatically searches for the optimal division of the family into subfamilies. The second method looks for positions whose mutational behavior is reminiscent of the mutational behavior of the full-length proteins, by directly comparing the corresponding distance matrices. The third method is an automation of the analysis of distribution of sequences and amino acid positions in the corresponding multidimensional spaces using a vector-based principal component analysis. These three methods have been tested on two non redundant lists of protein families: one composed by proteins that bind a variety of ligand groups, and the other composed by proteins with annotated functionally relevant sites. In most cases, the residues predicted by the three methods show a clear tendency to be close to bound ligands of biological relevance and to those amino acids described as participants in key aspects of protein function. These three automatic methods provide a wide range of possibilities for biologists to analyze their families of interest, in a similar way to the one presented here for the family of proteins related with ras-p21. PMID- 12589770 TI - SPACRCAN in the interphotoreceptor matrix of the mouse retina: molecular, developmental and promoter analysis. AB - SPACRCAN is a novel proteoglycan present in the interphotoreceptor matrix (IPM) of the rat and human retina that resists aqueous extraction through its binding to hyaluronan. The purpose of this study was: to clone mouse Spacrcan; to characterize the promoter elements; to define the deduced amino acid sequence; to establish the time of Spacrcan expression during retinal development; and to determine the time of appearance and distribution of SPACRCAN protein. Spacrcan cDNA clone was obtained through PCR amplification of a mouse retina cDNA library, and RT-PCR amplification and 5'RACE of mouse retina RNA. The deduced polypeptide sequence of mouse SPACRCAN contains a signal peptide at the N-terminal, seven N link glycosylation sites, numerous potential O-linked glycosylation sites in a central mucin-like domain, two glycosaminoglycan attachment sites, five potential hyaluronan-binding motifs, two epidermal growth factor-like domains, and a hydrophobic stretch of 23 amino acids near the C-terminal. Comparison of the genomic structure of mouse and human SPACRCAN showed significant structure conservation. Analysis of the promoter region revealed several important putative regulatory elements including a Ret-1/PCE-1 element, an 11 base motif for Crx binding, six copies of PIRE, a Ret-4 element, three copies of AP-1, a CRE element, and five copies of GATA3. Northern blot analysis and immunohistochemistry were used to determine the tissue specificity of Spacrcan mRNA and to localize SPACRCAN in developing retina. Spacrcan mRNA is expressed in both retina and pineal gland and was detectable as early as embryonic day 15. The protein is first detectable in the IPM at postnatal day 8 where it increases in concert with the extension of photoreceptor inner and outer segments from the outer retinal surface. The presence of several unique regulatory elements in the promoter region and characteristic molecular features shared with the orthologue in human and rat suggest an important functional role of SPACRCAN in the IPM. The time of appearance of the SPACRCAN protein during retinal development suggests that this matrix protein may establish the extracellular microenvironment into which photoreceptor outer segments are elaborated. PMID- 12589771 TI - Induction of citrulline-nitric oxide (NO) cycle enzymes and NO production in immunostimulated rat RPE-J cells. AB - Nitric oxide (NO) has been implicated in many physiological and pathological conditions in the eyes. The induction of inducible NO synthase (iNOS) and NO production have been noted in immunostimulated retinal pigment epithelial (RPE) cells. Cellular NO production depends on the availability of arginine, a substrate for NOS. Arginine can be regenerated from citrulline, another product of the NOS reaction, by argininosuccinate synthetase and argininosuccinate lyase, forming the citrulline-NO cycle. When rat RPE-J cells were treated with interferon-gamma (IFNgamma), tumor necrosis factor-alpha (TNFalpha) and lipopolysaccharide (LPS), and expression of the citrulline-NO cycle enzymes and related enzymes was analyzed, iNOS and argininosuccinate synthetase were highly induced at both mRNA and protein levels. On the other hand, argininosuccinate lyase was not induced. Among other related enzymes and transporters, mRNA for cationic amino acid transporter (CAT)-1 was weakly induced, whereas those for CAT 2, arginase I and II, ornithine aminotransferase and ornithine decarboxylase remained little changed. NO was produced by cells after stimulation with TNFalpha, IFNgamma and LPS. The induction of iNOS mRNA and the production of NO by these immunostimulated cells was further enhanced by cAMP. NO was produced from citrulline as well as from arginine. Our findings indicate that in activated RPE-J cells citrulline-arginine recycling is important for NO production. PMID- 12589772 TI - May nitric oxide molecule have a role in the pathogenesis of human cataract? AB - Recent studies have shown that nitric oxide molecule may have a role in the development of cataract. In this study, we measured the levels of a nitric oxide metabolite (nitrite) in the cataractous and normal human lenses. A modified Griess assay was used to determine the nitrite levels in the lenses as a measure of nitric oxide, based on the spectrophotometric method. Nitrite was detected in 26 (44.1%) cataractous lenses and was found below the detection limit in 33 (55.9%) cataractous lenses. Mean nitrite levels in cataractous lenses (2.77+/ 5.26nmol/100mg) was found higher than the normal lenses (0.77+/-0.79nmol/100mg) but this increase was not statistically significant. Comparison of nitrite levels among lenses with various types of cataracts revealed higher levels in lenses with posterior subcapsular cataracts. Hypertensive patients had also significantly higher nitrite levels in their cataractous lenses. The increased levels in the cataractous lenses display a possible role of nitric oxide in the pathogenesis of cataract in human eyes. PMID- 12589773 TI - Bi-directional migration of lens epithelial cells in a physiological electrical field. AB - An endogenous electric field (EF) exists in the vertebrate lens, but the influences of this on the functions of lens epithelial cells (LECs) are unclear. Because LECs from different regions experience different patterns of endogenous EFs and show striking differences in morphology and function, bovine LECs from different regions were cultured in an applied EF to mimic that occurring naturally. The migration of LECs from central and peripheral regions was both stimulated and directed by an applied EF. Field-directed cell migration required serum, or growth factors. Cells cultured in serum free medium lost directed migration completely and this was restored partially by the addition of basic fibroblast growth factor. The direction of cell migration depended on both EF strength and the origin of the LECs. Both central and peripheral LECs moved anodally at 150-250mVmm(-1). In a low EF (50mVmm(-1)), peripheral LECs migrated in the opposite direction, cathodally, but central LECs did not respond. Peripheral LECs alone therefore showed field strength dependent, bi-directional migration. A physiological EF enhanced activation of ERK1/2, a signaling molecule in the MAP kinase pathway, and this also required serum. Interestingly, expression of active ERK1/2 was enhanced in peripheral LECs at both 50 and 200mVmm(-1), which stimulated migration in different directions. In central LECs, which showed directed migration only at higher EFs, active ERK1/2 was increased only at 200mVmm(-1). The MAP kinase inhibitor U0126 prevented activation of ERK1/2 in LECs and inhibited EF-directed migration, implicating MAP kinase signaling in directing the migration of LECs in a physiological EF. PMID- 12589774 TI - The role of protein kinase C in modulation of aqueous humor outflow facility. AB - The elevated intraocular pressure that is commonly associated with glaucoma is believed to arise due to impairment of trabecular meshwork (TM) function. Although the TM and Schlemm's canal (SC) comprise the major route for aqueous humor outflow, little is known about the potential signaling mechanisms involved in the regulation of aqueous outflow. Based on knowledge regarding the role of protein kinase C (PKC) in vascular biology, we sought to understand the contribution of the PKC pathway towards outflow function by studying the modulation of contractile and morphological characteristics of TM and SC cells. We investigated the involvement of PKC in regulation of myosin light chain (MLC) phosphorylation, formation of actin stress fibers and integrin-ECM adhesions (focal adhesions) in human TM and SC cells and correlated these changes with aqueous outflow facility measured in an enucleated porcine whole eye perfusion model. Expression and distribution of PKC isoforms (alpha and epsilon ) in TM and SC cells and tissues was confirmed by Western blot and immunohistochemical analysis, respectively. Both, pharmacological activators (phorbol-12-myristate 13 acetate (PMA) and phorbol-12,13-dibutyrate (PDBu)) and inhibitors (staurosporine and GF109203X) of PKC were found to induce changes in cell shape (retraction and rounding up) and cytoskeletal organization in human TM and SC cells. While PMA and PDBu produced an increase in formation of actin stress fibers and focal adhesions and in MLC phosphorylation, PKC inhibitors were observed to induce contrasting effects in these cells. Intriguingly, both PDBU and GF109203X caused increases in aqueous outflow facility in the perfusion model. The PKC inhibitor (GF109203X) increased outflow by 46% while the PKC activator (PDBu) only increased outflow by 27%. These results suggest that PKC might play an important role in modulation of aqueous outflow facility by regulating MLC phosphorylation and thereby, the morphological and cytoskeletal characteristics of TM and SC cells. PMID- 12589775 TI - Propyl gallate is a superoxide dismutase mimic and protects cultured lens epithelial cells from H2O2 insult. AB - n-Propyl gallate (nPG) is a food preservative that is generally regarded as safe by the US FDA. It suppresses oxidation in biological systems. The mechanism by which nPG acts in biological systems is uncertain. We investigated whether nPG protected cultured lens epithelial cells from H(2)O(2)-induced damage. Cells were treated with H(2)O(2) or with nPG and then H(2)O(2). H(2)O(2) inhibited growth, caused membrane blebbing, decreased lactate production, increased the level of GSSG, decreased the levels of GSH, ATP and NAD(+), and G3PDH activity, stimulated the hexose monophosphate shunt and induced single-strand breaks in DNA. nPG prevented the H(2)O(2)-induced growth inhibition, membrane blebbing, drop in NAD(+) and single-strand breaks in DNA. The mechanism by which nPG acts at the chemical level was investigated using electron paramagnetic resonance (EPR), direct spectrophotometric kinetic measurements, and cyclic voltammetry. When nPG at low concentrations (nM to microM) was mixed with a large excess of O(2)(-)*, the superoxide signal was destroyed as indicated by UV visible spectroscopy and EPR. Kinetic analysis indicated that nPG dismutated O(2)(-)* in repetitive additions of superoxide with little loss of activity. The rate constant for the overall reaction of nPG with O(2)(-)* was ca. 10(6)M(-1)s(-1). nPG had a very low specific binding constant for Fe(2+) as determined by cyclic voltammetry. The evidence indicates that nPG dismutates the superoxide ion in a catalytic manner. PMID- 12589776 TI - Beta-adrenergic stimulation of Na(+)-K(+)-2Cl(-) cotransport activity in the rabbit lens. AB - Experimental maneuvers known to increase cellular cAMP levels evoked a stimulation in the K(+) influx across the anterior surfaces of isolated rabbit lenses, as measured by 86Rb(+) uptake. For this, the lenses were mounted in a modified Ussing-type chamber and exposed to the radiolabel under short-circuit conditions. The enhanced, cAMP-elicited flux was attributed to the basolateral Na(+)-K(+)-2Cl(-) cotransporter given its preclusion by bumetanide, a highly selective inhibitor of this symport, and the ineffectiveness of ouabain in mitigating the stimulation. The ouabain- plus bumetanide-insensitive K(+) uptake, which is about 10% of the total influx and represents passive entry of the radiolabel, was not affected by cAMP-elevating conditions. Forskolin, an activator of adenylyl cyclase; epinephrine, a non-selective adrenergic agonist; and the beta-selective agents, isoproterenol and terbutaline, were among the drugs used to elicit the increase in bumetanide-sensitive K(+) inflow. In experiments with isoproterenol, the stimulated influx evoked by the agonist was inhibited in lenses simultaneously exposed to propranolol. Other observations included that the stimulation of bumetanide-sensitive K(+) influx with forskolin was eliminated in lenses pretreated with the protein kinase inhibitors, staurosporine or H-89. However, these drugs were ineffective in preventing the increased influx produced by calyculin A, a phosphatase inhibitor, suggesting modulation of the cotransporter by at least two independent pathways. The cAMP generating stimuli also produced an inhibition of the short-circuit current across the lens and an increase in translens resistance. These latter effects suggest that cAMP elevation also evokes an inhibition in an epithelial conductance(s) simultaneously to the stimulation of the cotransporter. As such, this study provides the first indication for the regulation of lens transport by adrenoceptors, presumably of the beta-2 subtype. PMID- 12589777 TI - Apoptosis, necrosis, proliferation, and myofibroblast generation in the stroma following LASIK and PRK. AB - The aim of this study was to semi-quantitatively analyze stromal cell apoptosis, stromal cell proliferation, and myofibroblast generation over time points from 4hr to 3 months in rabbit eyes having photorefractive keratectomy (PRK) or laser in situ keratomeliusis (LASIK). Stromal cell necrosis and inflammatory cell infiltration were also studied. PRK for low myopia (-4.5diopters [D]), PRK for high myopia (-9.0D), and LASIK for high myopia (-9.0D) were performed in rabbit eyes, and corneas were obtained for examination at 4, 24, and 72hr, 1 and 4 weeks, and 3 months after surgery. A total of 144 rabbits were included in the study. Stromal cell apoptosis, proliferation, and myofibroblast generation were evaluated semi-quantitatively by TUNEL assay, immunocytochemical analysis of Ki67, and immunocytochemical analysis of alpha-smooth muscle actin, respectively. Stromal cell necrosis and characteristics of other cell types in the stroma were evaluated by electron microscopy. Keratocyte apoptosis and the subsequent proliferation and generation of myofibroblasts were qualitatively and quantitatively different in PRK for high myopia compared to either PRK for low myopia or LASIK for high myopia. Stromal cell necrosis becomes a significant form of cell death by 24hr after injury and may involve corneal fibroblasts, myofibroblasts, and inflammatory cells. Large numbers of polymorphonuclear cells and monocytes invade the cornea by 24hr after surgery and persist for over 1 week. The qualitative and quantitative differences in the cellular wound healing response after PRK for high and low myopia and LASIK for high myopia are likely determinants of the clinical differences in refractive outcome and some of the complications, such as regression and haze, seen after these procedures. PMID- 12589778 TI - Loss of melanin from human RPE with aging: possible role of melanin photooxidation. AB - The pigment melanin, which is believed to play a photoprotective role, was quantified here in human RPE cells from donors of different age. Electron spin resonance (ESR) spectroscopy was shown to provide a quantitative measure of melanin and was used as a non-destructive measure of melanin content. Results indicated an age-related melanin loss in RPE cells, with melanin content diminishing 2.5-fold between the first and the ninth decade of life. To determine whether photo-oxidation may contribute to age-related changes in RPE melanin, RPE in human eyecups, isolated human and bovine RPE cells, purified melanin granules, or synthetic dopa melanin were irradiated with various wavelengths and intensities of visible light. Samples were analysed for changes in melanin content by ESR spectroscopy, and by absorption and emission spectrophotometry. The concentration of hydrogen peroxide was measured in some samples, and some human eyecups were examined by transmission electron microscopy. Irradiation of RPE in eyecups with intense visible light was found to produce a time-dependent photobleaching of melanosomes that was accompanied by the formation of hydrogen peroxide. Photobleaching of isolated RPE melanosomes and synthetic dopa melanin resulted in enhanced melanin fluorescence, as previously shown for melanin from aged donors by others, and significantly reduced ESR signal intensity, resembling the changes in melanin with aging observed here. We conclude that the content of melanin in RPE cells undergoes an age-related change to which photo-oxidation may contribute. This observation raises the question of whether age-related changes in melanin reduce the photoprotective role of the pigment in aging RPE cells. PMID- 12589779 TI - Differential distribution of Mel(1a) and Mel(1c) melatonin receptors in Xenopus laevis retina. AB - The hormone melatonin is an output signal of an endogenous circadian clock in retinal photoreceptors. Melatonin may act as a paracrine and/or intracrine neurohormone by binding to specific receptors in the eye. The distribution of Mel(1a) and Mel(1c) melatonin receptors in the Xenopus laevis retina was examined by immunocytochemistry, using antibodies prepared against specific sequences of the Xenopus receptor proteins. Antibodies that label dopaminergic and GABA-ergic amacrine cells were used in double-label experiments with the melatonin receptor antibodies. The distribution of Mel(1a) and Mel(1c) receptor immunoreactivity was similar insofar as the two receptors were localized in the inner plexiform layer. However, the Mel(1c) receptor displayed some immunoreactivity in the photoreceptor cells, whereas the Mel(1a) receptor displayed little if any photoreceptor labelling. The Mel(1c) antibody, but not the Mel(1a), labelled a population of ganglion cells. While both receptors were localized to the outer plexiform layer, they did not appear to localize to the identical cell types. These results demonstrate that the Mel(1a) and Mel(1c) receptor proteins are present in cells of the X. laevis retina, and their distribution in the photoreceptors and inner retina is very similar to that reported in the human retina. The differential pattern of expression of the melatonin receptors suggests that melatonin may convey differential effects on various target cells in the retina. PMID- 12589780 TI - IL-4-induced cell proliferation and production of extracellular matrix proteins in human conjunctival fibroblasts. AB - Giant papillae, characteristic lesions of vernal keratoconjunctivitis, are formed as a result of the proliferation of conjunctival fibroblasts, the deposition of extracellular matrix, and the infiltration of inflammatory cells. The concentration of interleukin (IL)-4 is also increased in the tear fluid of individuals with ocular allergic diseases. The possible role of IL-4 in the development of giant papillae was investigated by examining the effects of this cytokine on cultured human conjunctival fibroblasts. Reverse transcription and polymerase chain reaction analysis revealed the presence of transcripts encoding the IL-4 receptor alpha chain in these cells, and flow cytometry demonstrated the expression of this protein on the cell surface. IL-4 induced the proliferation of conjunctival fibroblasts in a concentration-dependent manner, and this effect was inhibited by neutralizing antibodies to the IL-4 receptor. Enzyme immunoassays revealed that IL-4 also increased in a concentration-dependent manner the amounts of procollagen type I C-peptide and fibronectin released into the culture supernatant by conjunctival fibroblasts. A whole-cell enzyme-linked immunosorbent assay showed that IL-4 increased the deposition of collagen type III by conjunctival fibroblasts. Furthermore, reverse transcription combined with real time polymerase chain reaction analysis revealed that IL-4 increased the abundance of collagen type III mRNA in these cells. These results demonstrate that human conjunctival fibroblasts express receptors for IL-4, and that IL-4 stimulates both the proliferation of and the production of extracellular matrix proteins by these cells. These effects of IL-4 might contribute to the formation of giant papillae in individuals with vernal keratoconjunctivitis. PMID- 12589781 TI - Role of apoptosis in eyelid development. AB - The upper and lower eyelids fuse together during development and then reseparate. The mechanism of reseparation of the eyelids is still unclear, although many reports agree that keratinization has a major role in lid separation. We applied apoptotic detection methods with ultrastructural features to examine the process of lid separation in rats. We confirmed that the eyelids fused on the 18th day of gestation. At this stage, cellular excrescences appeared from the external and conjunctival surfaces overlying the junctional zone, and the junctional zone consisted of epidermal cell clumps with wide intercellular spaces. From the 18th to the 20th day of gestation, many apoptotic cells appeared in the junctional zone and in the cellular excrescences. In these tissues, TUNEL-positive cells and immunohistochemically stained macrophages were found at this stage. We conclude that apoptosis may eliminate external and conjunctival cellular excrescences to make both surfaces smooth and continuous across the junctional zone. Apoptosis may also play a major role in providing space for proliferation of newly organized epidermal cells without wide intercellular spaces. Subsequent keratinization extends onto epidermal and conjunctival sides of the junctional zone until separation of the lids is complete. PMID- 12589783 TI - Intense light exposure changes the crystallin content in retina. AB - Toward a better understanding of light-induced photoreceptor damage, the crystallin content of rat retina was examined following intense light exposure. Nine crystallin species were identified by mass spectrometric analysis of rat retina fractionated by 2D gel electrophoresis. The Coomassie blue staining intensity of all crystallin 2D gel components was 2- to 3-fold greater in light exposed than in control retinas. Following light exposure, anti-alphaB-crystallin immunoreactivity was increased in rod outer segments and retinal pigment epithelium. These findings support a possible role for crystallins in protecting photoreceptors from light damage. PMID- 12589782 TI - Lack of p75 receptor does not protect photoreceptors from light-induced cell death. AB - Rod photoreceptors are susceptible to light-induced cell death. Previous results have suggested that the neurotrophin receptor p75 in Muller cells controls photoreceptor cell death during light-exposure by suppressing trophic factor release; and consequently, if p75 is blocked or eliminated during light-exposure, apoptosis is delayed. We explored this question by examining photoreceptor cell survival in albino p75(-/-) mice as well as their heterozygous and homozygous littermates. Photoreceptor cell death was examined in semi-thin sections by counting the remaining rows of photoreceptors. No difference in the amount of cell death was found between p75(+/+) and p75(-/-) animals, whereas the single copy of p75 in the heterozygous p75(+/-) mice provided significant neuroprotection. Cell death in the wild-type animals may indeed be mediated by p75, whereas other known apoptosis pathways may be activated in the p75(-/-) mice. The pro-apoptotic activity of the p75 receptor may have been partially suppressed in the heterozygous p75(+/-) mice by the silencing effect of the Trk receptor. Thus, our results suggest that p75 signaling does not mediate the main apoptosis pathway activated during light-damage. PMID- 12589784 TI - Breaking the mould: archaea with all four chaperoning systems. PMID- 12589785 TI - Conversion of Ser to Thr residues at the sperm combining-site of mZP3 does not affect sperm receptor activity. AB - Mammalian eggs are surrounded by a thick extracellular coat, the zona pellucida, that is composed of three glycoproteins, called ZP1-3. Sperm recognize and bind to O-linked oligosaccharides attached to Ser-332 and Ser-334 at the sperm combining-site of mouse ZP3 (mZP3). Mutation of either of these Ser residues to a small aliphatic amino acid results in the loss of sperm binding to mZP3 in vitro. Here, we converted both Ser-332 and Ser-334 to Thr residues by site-directed mutagenesis. Recombinant mutant glycoprotein made by stably transfected EC cells was purified and then assayed for its ability to inhibit binding of sperm to ovulated eggs in vitro. Results of these experiments suggest that Thr residues can replace the two evolutionarily conserved Ser residues as acceptors for essential O-linked oligosaccharides at the sperm combining-site of mZP3 without affecting the glycoprotein's sperm receptor activity. PMID- 12589786 TI - Sequence analysis, expression, and paratope characterization of a single-chain Fv fragment for the eukaryote ribosomal P proteins. AB - The variable genes of monoclonal antibody (mAb) B10, specific for the C-terminal region of the eukaryotic ribosomal P protein, have been cloned as a single-chain Fv fragment (scFv) and expressed in Escherichia coli. The primary sequence of the variable regions of the B10 antibody, together with a detailed characterization of the reactive residues of the antigen, allowed the construction of a model of the paratope-epitope interaction, giving a first insight into the binding mechanisms of anti-P autoantibodies to their target peptides. The mAb and scFv could be useful for extensive P protein detection since both recognize the highly conserved motif DDxGF. PMID- 12589787 TI - Vasoactive intestinal peptide inhibits IL-8 production in human monocytes. AB - Vasoactive intestinal peptide (VIP), a neuropeptide present in the lymphoid microenvironment, acts as a potent anti-inflammatory agent that inhibits the function of activated macrophages. VIP was shown to inhibit IL-6, TNFalpha, IL 12, chemokine, and nitric oxide production in endotoxin-activated macrophages. The present study reports the effect of VIP on IL-8 production by stimulated human monocytes. VIP inhibits IL-8 production in a dose- and time-dependent manner at the mRNA level. The specific VPAC1 receptor mediates the inhibitory effect of VIP. Two transduction pathways appear to be involved, a major cAMP independent pathway and a secondary cAMP-dependent pathway. Of obvious physiological significance is the fact that VIP, presumably through the inhibition of IL-8 production, dramatically reduces the monocyte-induced neutrophil chemotaxis, an important event in the pathogenesis of several inflammatory and autoimmune disorders. These findings support the proposed role of VIP as a key endogenous anti-inflammatory agent and describe a novel mechanism, i.e., the inhibition of the production of monocyte-derived IL-8. PMID- 12589788 TI - Two lysosomal membrane proteins, LGP85 and LGP107, are delivered to late endosomes/lysosomes through different intracellular routes after exiting from the trans-Golgi network. AB - Lysosomal membrane proteins are delivered from their synthesis site, the endoplasmic reticulum (ER) to late endosomes/lysosomes through the Golgi complex. It has been proposed that after leaving the Golgi they are transported either directly or indirectly (via the cell surface) to late endosomes/lysosomes. In the present study, we examined the transport routes taken by two structurally different lysosomal membrane proteins, LGP85 and LGP107, in rat 3Y1-B cells. Here we show that newly synthesized LGP85 and LGP107 are delivered to late endosomes/lysosomes via a direct route without passing through the cell surface. Interestingly, although LGP107 is delivered from the Golgi to early endosomes containing internalized horseradish peroxidase-conjugated transferrin (HRP-Tfn) en route to lysosomes, LGP85 does not pass through the HRP-Tfn-positive early endosomes. These results suggest, therefore, that LGP85 and LGP107 are sorted into distinct transport vesicles at the post-Golgi, presumably the trans-Golgi network (TGN), after which LGP85 is delivered directly to late endosomes/lysosomes, but significant fractions of LGP107 are targeted to early endosomes before transport to late endosomes/lysosomes. This study provides the first evidence that after exiting from the Golgi, LGP85 and LGP107 are targeted to late endosomes/lysosomes via a different pathway. PMID- 12589789 TI - Interleukin 15 induces the signals of epidermal proliferation through ERK and PI 3-kinase in a human epidermal keratinocyte cell line, HaCaT. AB - Interleukin 15 (IL-15) is a potent stimulator of proliferation and an inhibitor of apoptosis in lymphocytes. We attempted to elucidate the mechanism of IL-15 function in HaCaT keratinocytes. We found that 5-bromo-2(')-deoxyuridine incorporation increased in a dose-dependent manner with IL-15. This was blocked by MEK inhibitor U0126 or PI 3-K inhibitor LY294002. ERK1/2 and Akt phosphorylation by IL-15 were detected in a dose- and time-dependent manner. U0126 and LY294002 abolished ERK1/2 and Akt phosphorylation, respectively. DNA fragmentation and Annexin V binding accompanied by UVB-induced apoptosis were reduced by 30-50% with IL-15. Taken together, IL-15 induced cellular proliferation and had an anti-apoptotic effect on keratinocytes, in which ERK1/2 and Akt phosphorylation played crucial roles. The signal transduction pathways of IL-15 in keratinocytes were partially elucidated; they share a substantial part with growth signals induced by EGF. These results suggest a therapeutic approach to inflammatory skin diseases by controlling these signals. PMID- 12589790 TI - Protein kinase A mediates cAMP-induced tyrosine phosphorylation of the epidermal growth factor receptor. AB - An increase in the intracellular cAMP concentration induces tyrosine phosphorylation of the epidermal growth factor receptor (EGFR) followed by activation of extracellular signal-regulated kinases 1/2 (ERK1/2). In this report we demonstrate that these effects of cAMP are mediated via activation of protein kinase A (PKA). Chemical inhibition of PKA suppressed forskolin-induced EGFR tyrosine phosphorylation and ERK1/2 activation in PC12 cells. Furthermore, forskolin failed to induce significant tyrosine phosphorylation of the EGFR and ERK1/2 activation in PKA-defective PC12 cells. Forskolin-induced EGFR tyrosine phosphorylation was also observed in A431 cells and in membranes isolated from these cells. Phosphoamino acid analysis indicated that the recombinant catalytic subunit of PKA elicited phosphorylation of the EGFR on both tyrosine and serine but not threonine residues in A431 membranes. Together, our data indicate that activation of PKA mediates the effects of cAMP on the EGFR and ERK1/2. While PKA may directly phosphorylate the EGFR on serine residues, PKA-induced tyrosine phosphorylation of the EGFR occurs by an indirect mechanism. PMID- 12589791 TI - Promoter analysis of the human SLC7A7 gene encoding y+L amino acid transporter-1 (y+LAT-1). AB - The human SLC7A7 gene on chromosome 14q11.2 encodes the y+L amino acid transporter-1 (y+LAT-1) protein that transports, together with the 4F2hc cell surface antigen, cationic amino acids through the basolateral membrane of epithelial cells in the small intestine and kidney. The SLC7A7 gene comprises 11 exons, but the first two are not translated. Mutations in the coding region of the SLC7A7 gene cause a rare autosomal disorder, lysinuric protein intolerance (LPI). We have now investigated the expression levels and putative 5' promoter elements of the SLC7A7. The 5' region of the first untranslated exon contains no TATA-box, Inr elements nor other classical promoter elements, but has instead other putative transcription factor binding sequences. The E-box and AP-2 elements were able to bind proteins in HEK293 cells and adult kidney tissue extracts, but not in fibroblasts. Using transient transfection and luciferase reporter gene studies, we showed that the first two introns located in the untranslated region contained transcriptional enhancer elements. Northern blot analysis showed low and equal SLC7A7 mRNA levels in the control and LPI patient fibroblastoid and lymphoblast cells. PMID- 12589792 TI - Priming-induced localization of G(ialpha2) in high density membrane microdomains. AB - Subcellular fractionation of human neutrophils on linear sucrose density gradients was utilized to test the hypothesis that priming regulates the subcellular and sub-plasma membrane distribution of neutrophil G-protein subunits, G(ialpha2) and G(ialpha3), N-formyl peptide receptor, Lyn kinase and phospholipase C(beta2). G(ialpha2), but not G(ialpha3), moved from a lighter to a higher density plasma membrane fraction. Unoccupied N-formyl peptide receptors were found throughout the plasma membrane fractions and this distribution did not change with priming. In unprimed cells G(ialpha2) and its effector, phospholipase C(beta2), were segregated in different membrane compartments; priming caused G(ialpha2) to move to the compartment in which phospholipase C(beta2) resided. Thus, an important component of the mechanism of priming may involve regulation of the location of G-proteins and effector molecules in plasma membrane compartments where their abilities to couple may be enhanced. PMID- 12589793 TI - Intracellular capture of B7 in antigen-presenting cells reduces costimulatory activity. AB - CTLA-4 gene constructs were designed to express CTLA-4 exclusively in the endoplasmic reticulum (ER). Four different CTLA-4 gene constructs were transfected into HEK 293 (human embryonic kidney) and A20 (Balb/c mouse B lymphoma) cells. All constructs contained an ER retention signal and coded for CTLA-4 expression in the ER. One of the constructs, which contained the membrane part of CTLA-4, coded for an expression both on the cell surface and in the ER. Three of the expressed CTLA-4 types (including the ER-membrane-expressed form) caused a reduced surface expression of B7 in the A20 cells. Only constructs which allow dimerization of CTLA-4 showed this effect. It is assumed that intracellular CTLA-4 bound B7 and inhibited therefore the transport of B7 to the surface. The binding obviously caused also an enhanced degradation of the complexes because both proteins showed a low concentration in the transfected cell lines. CTLA-4 transfected and B7-reduced A20 cells showed a diminished costimulating activity upon T cells. This was demonstrated by a reduced proliferation of T cells from ovalbumin-immunized Balb/c mice, incubated with ovalbumin peptide-primed CTLA-4 transfected A20 cells. PMID- 12589794 TI - A naturally enhanced green fluorescent protein from magnificent sea anemone (Heteractis magnifica) and its functional analysis. AB - A novel fluorescent protein termed hmGFP homologous to the green fluorescent protein (GFP) from Aequorea victoria was cloned from the tentacles of sea anemone Heteractis magnifica by EST sequencing and analysis of cDNA library and followed by using RT-PCR. The sequence analysis suggested that the chromophore, consensus amino acids, and secondary structure of 11 beta-strands of hmGFP were similar to those of GFP from other species. The recombinant hmGFP protein with high purity was obtained by the fusion expression of pETTRX-hmGFP in Escherichia coli and subsequent purification. The pH sensitivity and fluorescence spectroscopy of recombinant hmGFP were characterized. The excitation spectrum of recombinant hmGFP has a rather wide major peak with a maximum at 490 nm and a shoulder at 420 nm, and its emission spectrum at 510 nm. The expression of hmGFP and the chimera IPL through hmGFP in CHO cells has shown that the fusion protein IPL through hmGFP has retained the normal membrane targeting of the IPL from Dasyatis akajei, as well as maintaining fluorescent properties similar to those of native hmGFP, suggesting a promising prospect of the application in biotechnology research for the new protein. PMID- 12589795 TI - The carboxy-terminal pleckstrin homology domain of ROCK interacts with filamin-A. AB - The small GTPase Rho and its effector ROCK/Rho-kinase regulate actin cytoskeletal reorganization through phosphorylation of the regulatory light chain of myosin II. We previously reported that ROCK co-purified with the actin-binding protein filamin-A from HeLa cells. Here, we show that the pleckstrin homology (PH) domain of ROCK, but not the kinase or coiled-coil domain, interacts with filamin-A. We also determined that the PH domain of ROCK binds to the carboxy-terminal region of filamin-A containing the last 24th repeat. ROCK co-localized with filamin-A at the protrusive cell membranes of HeLa cells. PMID- 12589796 TI - Regulation of mitochondrial morphology by membrane potential, and DRP1-dependent division and FZO1-dependent fusion reaction in mammalian cells. AB - Mitochondria are dynamic organelles that undergo frequent fission and fusion or branching. To analyze the mitochondrial fusion reaction, mitochondria were separately labeled with green or red fluorescent protein (GFP and RFP, respectively) in HeLa cells, and the cells were fused using hemagglutinating virus of Japan (HVJ). The resulting mixing of the fluorescent reporters was then followed using fluorescence microscopy. This system revealed that mitochondria fuse frequently in mammalian cells, and the fusion depends on the membrane potential across the inner membrane. The protonophore, carbonyl cyanide m chlorophenylhydrazone (CCCP), led to fragmentation of the mitochondria and inhibited the fusion reaction. Removal of CCCP recovered the fusion activity to reform filamentous mitochondrial networks. Analysis of the effects of GTP-binding proteins, DRP1 and two FZO1 isoforms, and the GTPase-domain mutants on the CCCP induced mitochondrial morphologic changes revealed that DRP1 and FZO1 are involved in membrane budding and fusion, respectively. Furthermore, a HVJ dependent cell fusion assay combined with RNA interference (RNAi) demonstrated that both FZO1 isoforms are essential and must be acting in cis for the mitochondrial fusion reaction to occur. PMID- 12589797 TI - Differential control of the NIMA-related kinases, Nek6 and Nek7, by serum stimulation. AB - Neks (NIMA-related kinases) are mammalian serine/threonine (Ser/Thr) protein kinases structurally related to Aspergillus NIMA (Never in Mitosis, gene A), which plays essential roles in mitotic signaling. Among these kinases, Nek6 and Nek7 are structurally related and constitute a subfamily in the NIMA/Nek family, although their functions still remain almost elusive. In this report, we studied the enzymatic regulation of Nek6 and Nek7 to gain an insight into their cellular functions. Recombinant Nek7 produced in bacteria was active comparably to Nek6; however, the Nek7 activity in mammalian cells was found to be significantly lower than Nek6. Since Nek6 previously has been reported to in vitro phosphorylate p70 ribosomal S6 kinase at Thr412, we examined if Nek6 and Nek7 activities were controlled by the amino acid supplement, which is known to affect the phosphorylation at Thr412, and did not observe any significant effect. However, we unexpectedly found that Nek7 kinase activity was rapidly and efficiently increased by serum deprivation, while Nek6 activity was decreased. This is well consistent with the lower activity of Nek7 in cells under normal growth conditions. In addition, it was suggested that Nek7 activity would be regulated in a cell cycle-dependent manner, although Nek6 was not. These clear differences in enzymatic control between the highly similar kinases, Nek6 and Nek7, suggest their distinct signaling functions in mammalian cells. PMID- 12589798 TI - Role of calcium in E-selectin induced phenotype of T84 colon carcinoma cells. AB - The adhesion of cancer cells to the endothelium during the metastatic process involves the interaction of specific cell-cell adhesion receptors on the cell surface. E-selectin on endothelial cells and sialyl Lewis X carbohydrate component on tumor cells are mainly implicated in the adhesion of colon carcinoma cells to the endothelium of target organ. In this paper we show that binding of E selectin to T84 colon tumor cells causes approximately a twofold increase in intracellular calcium concentration. In particular, using two inhibitors of receptor operated calcium channels, CAI and SK&F 96365, we present evidences that the augmentation in cytoplasmic calcium originates from ionic influx from extracellular sources. Furthermore, we demonstrated that modulation of [Ca2+]i by engagement of E-selectin receptor starts signal transduction pathways that affect cell spreading, tyrosine phosphorylation signaling, and cancer cell motility. PMID- 12589799 TI - Transcription of ahpC, katG, and katE genes in Escherichia coli is regulated by polyamines: polyamine-deficient mutant sensitive to H2O2-induced oxidative damage. AB - Polyamines (putrescine and spermidine) are present in almost all living organisms and participate in numerous cellular processes. In this study, we report the protective roles of polyamines against hydrogen peroxide (H2O2)-induced oxidative stress. All of ahpC, katG, and katE genes, known to participate in the antioxidant defense mechanism against H2O2-induced stress in Escherichia coli, failed to induce in the absence of polyamines during normal aerobic growth. The induction of both oxyR and rpoS gene expression, whose products are essential to induce ahpC, katG, and katE genes, was also absolutely dependent on polyamines. Polyamine-deficient E. coli mutant has increased susceptibility to exogenous H2O2, and this cell cytotoxicity was relieved to a wild-type level by addition of putrescine or spermidine (1mM), which restored the transcriptional induction of ahpC, katG, and katE genes. H2O2-removing capacity was measured in the mutant, showing a significantly low H2O2-removing capacity compared to the wild type when polyamines were not present. We concluded that the increased susceptibility of the polyamine-deficient E. coli mutant to H2O2 treatment resulted from an intracellular low level of H2O2-removing capacity through the failure of their regulons, ahpC, katG, and katE induction, as well as the failure of oxyR and rpoS induction. PMID- 12589800 TI - UV-light induces p38 MAPK-dependent phosphorylation of Bcl10. AB - Bcl10 is a signaling protein required for activation of the NF-kappaB transcription factor. NF-kappaB is an important mediator of genotoxic stress and regulates the expression of genes required for both cell proliferation and cell death. Bcl10 is phosphorylated in vivo, however, the protein kinase or kinases responsible are not known. Here, we show that Bcl10 is phosphorylated in response to UV irradiation. UV-induced phosphorylation of Bcl10 was inhibited by the p38 stress-activated protein kinase inhibitors SB203580 and PD169316, suggesting that p38 is required for UV-mediated phosphorylation of Bcl10. PMID- 12589801 TI - Direct interaction of the Golgi membrane with the endoplasmic reticulum membrane caused by nordihydroguaiaretic acid. AB - Nordihydroguaiaretic acid (NDGA), an inhibitor of lipoxygenase, blocks protein transport from the endoplasmic reticulum (ER) to the Golgi complex and induces the redistribution of Golgi proteins into the ER. We investigated characteristics of NDGA-induced retrograde movement of the Golgi proteins to the ER. At an early stage of incubation of cells with NDGA, the Golgi complex formed convoluted membrane aggregates. Electron microscopy revealed that these aggregates directly interact en bloc with the ER membrane. The direct interaction and subsequent incorporation of the Golgi proteins into the ER were found to be temperature dependent. The protein of ER-Golgi intermediate compartment (ERGIC), ERGIC53, was rapidly accumulated in the Golgi upon treatment with NDGA. This accumulation was significantly inhibited by low temperature at 15 degrees C. Under the condition, the redistribution of the Golgi proteins into the ER as well as the direct interaction between the ER and the Golgi by NDGA were also inhibited, suggesting an important role of the ERGIC in the retrograde movement. In contrast, the low temperature did not inhibit formation of the Golgi aggregates by NDGA. Taken together, these results suggest that NDGA causes the redistribution of the Golgi proteins into the ER through the direct connections between the Golgi, the ERGIC, and the ER. PMID- 12589802 TI - Activation of Rac and tyrosine phosphorylation of cytokine receptors induced by cross-linking of integrin alpha4beta1 and cell adhesion in hematopoietic cells. AB - Adhesion of hematopoietic cells, mainly through alpha4beta1 and alpha5beta1 integrins, to the bone marrow microenvironment may play important roles in regulation of hematopoiesis. However, the mechanisms for signaling, outside-in signaling, have largely remained to be established. We demonstrate here that cross-linking of alpha4beta1 by anti-alpha4 antibody induces tyrosine phosphorylation of Pyk2, Shc, and Cbl as well as binding of the adaptor protein CrkL with Cbl in a murine hematopoietic cell line, 32D/EpoR-Wt. Furthermore, cross-linking of alpha4beta1 induced activation of the Rho family small GTPase Rac, which was enhanced by induced overexpression of CrkL and was inhibited by the phosphatidylinositol 3(')-kinase (PI3K) inhibitor LY294002. In addition, adhesion of 32D/EpoR-Wt cells to immobilized H-296, a recombinant fibronectin peptide specific for alpha4beta1, induced tyrosine phosphorylation of Jak2, the erythropoietin receptor (EpoR), and the IL-3 receptor beta subunit as well as Pyk2, Shc, and Cbl. Tyrosine phosphorylation of Jak2 and EpoR was also induced in a human leukemic cell line, UT-7, by adhesion to immobilized H-296. However, adhesion of 32D/EpoR-PM4 cells, expressing the W282R mutant EpoR defective in coupling with Jak2, to immobilized H-296 failed to induce tyrosine phosphorylation of the mutant EpoR. These results implicate CrkL in PI3K dependent activation of Rac by outside-in signaling from alpha4beta1 and suggest that adhesion through alpha4beta1 further activates cytokine receptor-associated Jak2 to induce phosphorylation of these receptors. PMID- 12589803 TI - The Drosophila SSL gene is expressed in larvae, pupae, and adults, exhibits sexual dimorphism, and mimics properties of the beta subunit of casein kinase II. AB - Drosophila melanogaster casein kinase II (CKII) is composed of catalytic alpha and regulatory beta subunits that generate the alpha2beta2 holoenzyme. A two hybrid screen of a Drosophila embryo library using CKIIalpha as bait has resulted in the isolation of multiple cDNAs encoding SSL, a CKIIbeta-like polypeptide. We demonstrate that CKIIbeta, beta', and SSL exhibit robust and comparable interaction with CKIIalpha. Residues in SSL that mediate interaction with CKIIalpha appear similar to those in CKIIbeta, and SSL forms homodimers and heterodimers with CKIIbeta or beta' as well. We have tested all known Drosophila CKIIbeta-like proteins for rescue of the ion-homeostasis defect of yeast lacking beta subunits and find that CKIIbeta and SSL complement, beta' has marginal function, and Stellate appears non-functional. We have used real-time RT-PCR to assess developmental expression, and find that CKIIbeta is robust and ubiquitous, whereas SSL is restricted to males (third-instar-larvae, pupae, and adults), but is nondetectable in females of the corresponding stages. These results indicate that SSL expression encompasses a greater developmental window than that previously suggested and may confer distinct functions to CKII in a sex-specific manner. PMID- 12589804 TI - Acylphosphatase interferes with SERCA2a-PLN association. AB - We previously reported that acylphosphatase, a cytosolic enzyme present in skeletal and heart muscle, actively hydrolyzes the phosphoenzyme (EP) of cardiac sarcoplasmic reticulum (SR) Ca2+-ATPase (SERCA2a), inducing an increased activity of this pump. We hypothesized that acylphosphatase-induced stimulation of SERCA2a, in addition to enhanced EP hydrolysis, may be due to a displacement of phospholamban (PLN), removing its inhibitory effect. To verify this hypothesis co immunoprecipitation experiments were performed by adding recombinant muscle acylphosphatase to solubilized heart SR vesicles, used as a source of SERCA2a and PLN. With anti-acylphosphatase antibodies only SERCA2a was co-immunoprecipitated in an amount which increased in parallel to the concentrations of our enzyme. Conversely, using anti-SERCA2a antibody, both PLN and acylphosphatase were co immunoprecipitated with SERCA2a, and the PLN amount in the precipitate decreased with increasing acylphosphatase concentrations. SERCA2a and PLN were co immunoprecipitated by anti-phospholamban antibodies, but while the amount of precipitated phospholamban increased in the presence of acylphosphatase, the level of SERCA2a decreased. These preliminary results strengthen the supposed displacement of phospholamban by acylphosphatase. PMID- 12589805 TI - The depletion of protein signals in metabonomics analysis with the WET-CPMG pulse sequence. AB - Nuclear magnetic resonance (NMR) spectroscopy is a powerful analytical tool capable of providing a comprehensive metabolic profile of biofluids such as urine, plasma, and serum. Unfortunately, when measuring serum and plasma, the high protein concentration can obscure the signals originating from low molecular weight metabolites. We evaluated the use of different parameters within the Carr Purcell-Meiboom-Gill (CPMG) pulse train of fast spin-echoes to remove the macromolecular signal contribution in one-dimensional proton (1H) NMR spectra. Experimental parameters such as the refocusing delay in the CPMG pulse train, pulse miscalibration, and recycle time were examined to assess the ability to remove the protein signals from the spectrum without causing a deleterious effect on the signals originating from free, low molecular weight metabolites. The 1H NMR spectra of a variety of serum samples spiked with 2'-deoxyadenosine were acquired using various acquisition parameters. Our results show that the delay used in the CPMG spin-echo and the combination of the acquisition pulse flip angle and recycle time are the two major factors affecting the observed metabolite signal amplitudes in the resulting 1H-NMR spectrum. PMID- 12589806 TI - Cloning and phylogenetic analysis of an amphioxus myogenic bHLH gene AmphiMDF. AB - In this study, a member of the MyoD gene family, AmphiMDF, was isolated from the embryos of amphioxus by degenerate PCR, followed by rapid amplification of cDNA ends (RACE). Southern blot analysis confirmed that only a single myogenic bHLH gene was present in the genome of amphioxus Branchiostoma belcheri tsingtauense. Sequence and phylogenetic analyses indicated that AmphiMDF falls at the base of its vertebrate homologs. The amino acid sequence of AmphiMDF was almost equally similar to those of the four clusters of the vertebrate MyoD family. This suggests that AmphiMDF is not only the sister but also the archetype of the vertebrate myogenic bHLH genes. The scenarios to explain the origin of the vertebrate MyoD gene family from the ancestral myogenic bHLH gene like AmphiMDF are also discussed. PMID- 12589807 TI - Apoptosis induced by butyrate is independent of Jak/STAT signaling in a fibrosarcoma cell line. AB - The aim of this study was to evaluate the participation of the Jak-1 and STAT-1 proteins in sodium butyrate-induced apoptosis in 2C4 cells derived from human fibrosarcoma. Making use of Jak-1 or STAT-1 deficient cell lines, we demonstrated that the apoptotic process induced by butyrate is independent of the presence of these proteins. In addition, this work showed that, although the constitutive expression of pro-caspases-2 and -3 is reduced in STAT-1 cells, the activity of caspase-3 is preserved in both Jak-1 and STAT-1 deficient cells and is similar to that seen in 2C4 parental cells. In conclusion, we demonstrated that the absence of functionally active Jak-1 or STAT-1 protein directly affects the TNF-alpha induced apoptosis, but does not alter the sodium butyrate-induced apoptosis in cells derived from human fibrosarcoma. PMID- 12589808 TI - A novel T7 system utilizing mRNA coding for T7 RNA polymerase. AB - The T7 system dose not require the relocation of a reporter gene to the nucleus for its gene expression in the cytoplasm, but relies on the co-localization of T7 RNA polymerase (T7 RNAP) enzyme and reporter gene DNA that is controlled by the T7 promoter. In the present study, we developed a new T7 system in that gene expression can occur at a higher level than those using conventional systems. Insertion of 5'- and 3'-untranslated regions (UTR) of beta-globin gene into a reporter gene enhanced the reporter gene expression, presumably due to the stability and efficient translation of the mRNA. Instead of the T7 RNAP protein used in conventional methods, moreover, transfection of cells with T7 RNAP mRNA, which has been modified by inserting beta-globin 5'- and 3'-UTR sequences as well as the cap and poly(A) tail structures, further enhanced the reporter gene expression. Thus, this novel T7 system using T7 RNAP mRNA may be powerful for the efficient gene expression of DNA exogenously provided in the cytoplasm. PMID- 12589809 TI - Phospholipid assisted folding of a denatured heme protein: effect of phosphatidylethanolamine. AB - The role of the aminophospholipid, phosphatidylethanolamine (PE), has been well established to act as a non-protein molecular chaperone in the folding and assembly of polytopic membrane proteins. However, such studies with soluble proteins have not been done so far and in particular with the heme proteins. We have used the heme enzyme, horseradish peroxidase (HRP), as the model heme protein and studied the effect of different phospholipids on its refolding from denatured state. Dimyristoylphosphatidylethanolamine (DMPE), a bilayer-forming PE, was able to increase the reactivation yield of denatured HRP upon 30min refolding at 25 degrees C. However, dioleoylphosphatidylethanolamine (DOPE), containing one double bond in the fatty acid chains, which does not favour bilayer organization, did not support proper refolding. The phospholipids with N methylated head groups, phosphatidylcholines, e.g., DMPC and DOPC showed differential effects when DMPC remained mostly non-supportive while DOPC on the contrary led to inhibition of the refolding of the denatured heme enzyme. Fluorescence spectroscopic studies also indicated changes in the microenvironments of the heme moiety and the single tryptophan residue of HRP in presence of the aminophospholipid. PMID- 12589810 TI - Imipenem and expression of multidrug efflux pump in Enterobacter aerogenes. AB - Imipenem is often used to treat intensive care unit patients infected by Enterobacter aerogenes, but it is leading to an increasing number of antibiotic resistant strains. Clinical isolates and imipenem resistant variants presented a high level of resistance to beta-lactam antibiotic group and to chemically unrelated drugs. We report here that imipenem selects strains which contain active efflux pumps ejecting various unrelated antibiotics including quinolones, tetracycline, and chloramphenicol. An increase of AcrA, an efflux pump component, was observed in the imipenem resistant variants. The overexpression of marA, involved in the genetic control of membrane permeability via porin and efflux pump expression, indicated the activation of the resistance genetic cascade in imipenem resistant variants. PMID- 12589811 TI - The interaction between ADAM 22 and 14-3-3zeta: regulation of cell adhesion and spreading. AB - The ADAM family consists of a number of transmembrane proteins that contain disintegrin-like and metalloproteinase-like domains. Therefore, ADAMs potentially have cell adhesion and protease activities. 14-3-3 proteins are a highly conserved family of cytoplasmic proteins that associate with several intracellular signaling molecules in the regulation of various cellular functions. Here we report the identification of a novel interaction between the ADAM 22 cytoplasmic tail and the 14-3-3zeta isoform by a yeast two-hybrid screen. The interaction between the ADAM 22 cytoplasmic tail and 14-3-3zeta was confirmed by an in vitro protein pull-down assay as well as by co-immunoprecipitation, and the binding sites were mapped to the 28 amino acid residues of the C-terminus of the ADAM 22 cytoplasmic tail. Furthermore, we found that overexpression of the ADAM 22 cytoplasmic tail in human SGH44 cells inhibited cell adhesion and spreading and that deletion or mutation of the binding site for 14-3-3zeta within the ADAM 22 cytoplasmic tail abolished the ability of the overexpressed cytoplasmic tail to alter cell adhesion and spreading. Taken together, these results for the first time demonstrate an association between ADAM 22 and a 14-3 3 protein and suggest a potential role for the 14-3-3zeta/ADAM 22 association in the regulation of cell adhesion and related signaling events. PMID- 12589812 TI - Multiple domains of the mouse p19ARF tumor suppressor are involved in p53 independent apoptosis. AB - The ARF (p19ARF for the mouse ARF consisting of 169 amino acids and p14ARF for the human ARF consisting of 132 amino acids) genes upregulate p53 activities to induce cell cycle arrest and sensitize cells to apoptosis by inhibiting Mdm2 activity. p53-independent apoptosis also is induced by ectopic expression of p19ARF. We constructed various deletion mutants of p19ARF with a cre/loxP regulated adenoviral vector to determine the regions of p19ARF which are responsible for p53-independent apoptosis. Ectopic expression of the C-terminal region (named C40) of p19ARF whose primary sequence is unique to the rodent ARF induced prominent apoptosis in p53-deficient mouse embryo fibroblasts. Relatively low-grade but significant apoptosis also was induced in p53-deficient mouse embryo fibroblasts by ectopic expression of p19ARF1-129, a p19ARF deletion mutant deficient in the C40 region. In contrast, ectopic expression of the wild-type p14ARF did not induce significant apoptosis in human cells. Taken together, we concluded that p53-independent apoptosis was mediated through multiple regions of the mouse ARF including C40, and the ability of the ARF gene to mediate p53 independent apoptosis has been not well conserved during mammalian evolution. PMID- 12589813 TI - The p75 receptor is required for BDNF-induced differentiation of neural precursor cells. AB - Epidermal growth factor (EGF)-treated neurospheres from fetal forebrain contain multipotential cells capable of neuronal, astrocytic, and oligodendroglial differentiation. These neural precursor cells express the TrkB as well as the neurotrophin receptor p75 (p75NTR), suggesting that they are BDNF responsive. In this study, we test whether the p75NTR plays a role in the differentiation of these neural precursor cells in vitro. Activation of the TrkB and the p75NTR by the addition of BDNF facilitates neuronal commitment and marked neurite genesis. However, no promotion of neuronal commitment by BDNF was observed in the neural precursor cells from mice carrying a mutation in the p75NTR gene. In addition, we observed a significant increase in the number of nestin-positive cells and the proliferation of the cells lacking functional p75NTR. These findings suggest that the p75NTR is required for proper neuronal fate decision as well as the differentiation of the neural precursor cells. PMID- 12589814 TI - Reconstitution of lethally irradiated mice by cells isolated from adipose tissue. AB - It is suggested that hematopoietic stem cells (HSC) could be found in several tissues of mesodermic origin. Among these, adipose tissue can expand throughout adult life and its expansion is not only due to mature adipocyte hypertrophy but also to the presence of precursor cells in stroma-vascular fraction (SVF). Here we report that transplantation of cells isolated from mice adipose tissue can efficiently rescue lethally irradiated mice and results in a reconstitution of major hematopoietic lineages. Donor cells can be detected in blood and in hematopoietic tissues of recipient mice. Adipose tissue contains a significant percentage of CD34, CD45 positive cells, and SVF cells were able to give rise to hematopoietic colonies in methylcellulose. We demonstrate the presence of hematopoietic progenitors in adipose tissue by phenotypic and functional characteristics. Thus adipose tissue could be considered as an important and convenient source of cells able to support hematopoiesis. PMID- 12589815 TI - Structural and enzymatic characterization of physarolisin (formerly physaropepsin) proves that it is a unique serine-carboxyl proteinase. AB - Previously, we purified and partially characterized physarolisin, a lysosomal acid proteinase from Physarum polycephalum, which had been suggested to be concerned with the morphological changes of the mold. In this study, a cDNA for the enzyme was cloned and sequenced, and the structural and enzymatic features were investigated. The enzyme shows a sequence similarity to the serine-carboxyl proteinase family (MEROPS S53). Indeed, diisopropylfluorophosphate (DFP) was shown to strongly inhibit the activity of the enzyme. However, the enzyme possesses several unique features distinct from the other members of the family, such as the two-chain structure and inhibition by diazoacetyl-D,L-norleucine methyl ester (DAN). The sites and mode of processing of the precursor to the mature enzyme were deduced, and the major DAN-reactive residue in the enzyme was identified to be Asp529. These features were suggested to be due to the unique local tertiary structure of the enzyme by molecular modeling. We now propose the name physarolisin for the enzyme. PMID- 12589816 TI - Induction of human mammary-associated serum amyloid A3 expression by prolactin or lipopolysaccharide. AB - In most mammalian species, serum amyloid A isoform 3 (SAA3) appears to be the predominant SAA isoform expressed extrahepatically. However, human SAA3 gene expression has not been detected previously and, therefore, this gene was referred to as a pseudogene. We report for the first time the transcriptional expression of human SAA3. Human SAA3 gene expression was detected by RT-PCR after stimulation of mammary gland epithelial cells with either prolactin (PRL) or lipopolysaccharide (LPS). The full-length 655bp cDNA sequence for this mammary associated serum amyloid A3 (M-SAA3) was obtained using 5' and 3' rapid amplification of cDNA ends (RACE). The human M-SAA3 transcript would conceptually translate into a 42 residue mature protein, which is smaller than other mammalian SAA3 isoforms that are typically 104-113 amino acids in length. This study defines the cDNA sequence for human SAA3 and also demonstrates the upregulation of M-SAA3 expression in response to the lactational hormone PRL or to an acute phase stimulant such as LPS. PMID- 12589818 TI - Adiponectin gene expression and secretion is inhibited by interleukin-6 in 3T3-L1 adipocytes. AB - Recently, it has been shown that adiponectin is an important insulin-sensitizing fat-derived protein which is downregulated in insulin resistance and obesity, and replenishment of which improves insulin sensitivity. In contrast, interleukin (IL)-6 appears as an adipocytokine serum concentrations of which are elevated in these states. However, it has not been determined whether IL-6 might impact on expression and secretion of adiponectin. To clarify this, 3T3-L1 adipocytes were treated with different concentrations of IL-6 for various periods of time. Adiponectin mRNA was measured by quantitative real-time reverse transcription polymerase chain reaction and secretion was determined by radioimmunoassays. Interestingly, treatment of 3T3-L1 cells with 30 ng/ml IL-6 significantly decreased adiponectin secretion to 75% of control levels. Adiponectin secretion was also inhibited between 25% and 45% by chronic treatment with forskolin (50 microM), tumor necrosis factor alpha (100 ng/ml), and dexamethasone (100 nM). Furthermore, adiponectin mRNA expression was downregulated by up to 50% in a time and dose-dependent manner, with significant inhibition detectable at concentrations as low as 3 ng/ml IL-6 and as early as 8h after effector addition. The inhibitory effect of IL-6 was partially reversed by pretreatment of 3T3-L1 cells with pharmacological inhibitors of a p44/42 mitogen-activated protein (MAP) kinase. Moreover, the negative effect of IL-6 on adiponectin mRNA expression could be reversed by withdrawal of the hormone for 24h. Taken together, our results suggest that adiponectin gene expression is reversibly downregulated by IL-6 and support the concept of adiponectin being an important selectively controlled modulator of insulin sensitivity. PMID- 12589817 TI - Immortalization with telomerase of the Nestin-positive cells of the human pancreas. AB - Cells expressing the neuronal stem cell marker Nestin are present in the human pancreas but the biological role of these cells has yet to be resolved. We report here the establishment with the catalytic subunit of human telomerase (hTERT) of a line of normal human cells representing this cell type. Primary human cells derived from the ducts of the pancreas were transduced with an hTERT cDNA. The infected cells became positive for telomerase, failed to senesce, and were still proliferating after more than 150 doublings. The immortalized cells were positive for the expression of Nestin (at both the mRNA and protein levels) and were found to be free of cancer-associated changes: diploid and expressing wild type p16(INK4a), p53, and K-Ras. An established line of normal human cells representing this cell type should be of great value to help define the biological properties of this novel cell type. PMID- 12589819 TI - Functional significance of the thyrotropin receptor germline polymorphism D727E. AB - In a toxic thyroid adenoma we identified a novel somatic mutation that constitutively activates the thyrotropin receptor (TSHR). Two heterozygous point mutations at adjacent nucleotides led to a substitution of alanine with asparagine at codon 593 (A593N) in the fifth transmembrane helix of TSHR. This somatic mutation resided on the same TSHR allele with the germline polymorphism D727E. The functional characteristics of the single TSHR mutants A593N and D727E and of the double mutant A593N/D727E were studied in transiently transfected COS 7 cells. The TSHR mutants A593N and A593N/D727E constitutively activated the cAMP cascade, whereas the D727E mutant did not differ from the wild-type TSHR. Surprisingly, the double mutant's specific constitutive activity was 2.3-fold lower than the A593N mutant. Thus, the polymorphism significantly ameliorates G(alphas) protein activation in the presence of the gain-of-function mutation A593N, although it is functionally inert in the context of the wild-type TSHR. PMID- 12589820 TI - Identification and characterization of two new human mu opioid receptor splice variants, hMOR-1O and hMOR-1X. AB - The mouse gene encoding the mu opioid receptor, Oprm, undergoes extensive alternatively splicing, with 14 variants having been identified. However, only one variant of human mu opioid receptor gene (Oprm), MOR-1A, has been described. We now report two novel splice variants of the human Oprm gene, hMOR-1O and hMOR 1X. The full-length cDNAs of hMOR-1O and hMO-1X contained the same exons 1, 2, and 3 as the original hMOR-1, but with exon O or exon X as the alternative fourth exon, respectively. Northern blots revealed several bands with the exon O probe in both human neuroblastoma BE(2)C cells and human brain and a single band (5.5kb) with the exon X probe in selected human brain regions. When transfected into CHO cells, both variants showed high selectivity for mu opioids in binding assays. These two new human mu opioid receptors are the first human MOR-1 variants containing new exons and suggest that the complex splicing present in mice may extend to humans. PMID- 12589821 TI - Inhibition of Epstein-Barr virus lytic cycle by (-)-epigallocatechin gallate. AB - (-)-Epigallocatechin gallate (EGCG), abundant in green tea, is a potent anti microbial and anti-tumor compound. This investigation used immunoblot, flow cytometry, microarray, and indirect immunofluorescence analyses to show that at concentrations exceeding 50 microM, EGCG inhibits the expression of Epstein-Barr virus (EBV) lytic proteins, including Rta, Zta, and EA-D, but does not affect the expression of EBNA-1. Moreover, DNA microarray and transient transfection analyses demonstrated that EGCG blocks EBV lytic cycle by inhibiting the transcription of immediate-early genes, thus inhibiting the initiation of EBV lytic cascade. PMID- 12589822 TI - Quercetin exerts multiple inhibitory effects on vascular smooth muscle cells: role of ERK1/2, cell-cycle regulation, and matrix metalloproteinase-9. AB - The French paradox has been attributed to the antioxidant properties of flavonoids present in the red wine. Quercetin, a bioflanoid present in the human diet, is known to inhibit angiotensin II-induced hypertrophy and serum-induced smooth muscle cell proliferation. However, it is not known whether quercetin exerts similar cardioprotective effects in cells treated with TNF-alpha. In this study, we investigated whether quercetin exerts the multiple suppressive effects on cytokine TNF-alpha-induced human aortic smooth muscle cells (HASMC). Treatment of quercetin showed potent inhibitory effects on the DNA synthesis of cultured HASMC in the presence of TNF-alpha. These inhibitory effects were associated with reduced extracellular signal-regulated kinase (ERK)1/2 activity and G1 cell-cycle arrest. Treatment of quercetin, which induced a cell-cycle block in G1-phase, induced down-regulation of cyclins and CDKs and up-regulation of the CDK inhibitor p21 expression, whereas up-regulation of p27 or p53 by quercetin was not observed. Because anti-atherogenic effects need not be limited to antiproliferation, we decided to examine whether quercetin exerted inhibitory effects on matrix metalloproteinase-9 (MMP-9) activity in TNF-alpha-induced HASMC. Quercetin inhibited TNF-alpha-induced MMP-9 secretion on HASMC in a dose dependent manner. This inhibition was characterized by down-regulation of MMP-9, which was transcriptionally regulated at NF-kappaB site and activation protein-1 (AP-1) site in the MMP-9 promoter. These findings indicate the efficacy of quercetin in inhibiting cell proliferation, G1- to S-phase cell-cycle progress, and MMP-9 expression through the transcription factors NF-kappaB and AP-1 on TNF alpha-induced HASMC. PMID- 12589823 TI - Molecular cloning and characteristics of a novel zinc finger protein and its splice variant whose transcripts are expressed during spermatogenesis. AB - Testicular zinc finger protein (TZF) has a zinc finger motif of the Cys2-His2 type and its transcript is expressed predominantly in mouse spermatogenic cells. Using the fragment of TZF as a probe, we isolated the alternative splice variant form (TZF-L) from mouse testis cDNA library. Analysis of the open reading frame of each cDNA indicated that TZF and TZF-L were polypeptides of 942 and 2025 amino acid residues, respectively, and the N-terminal 902 amino acids of TZF-L were identical to those of TZF. The C-terminal region of TZF-L had more a zinc finger motif of the Cys2-His2 type and poly-Glu and poly-Pro regions. The mouse TZF/TZF L gene spanned >20 kb and consisted of 11 exons. RT-PCR analysis of the expression level of mRNAs for mouse TZF and TZF-L showed that both transcripts are highly expressed in testis and moderately in kidney and ovary. Elevated expression of both transcripts during testicular development in mice was restricted to spermatocytes at the pachytene stage of meiotic prophase. Fusion proteins with GFP also demonstrated the nuclear localization of TZF and TZF-L. These experiments suggest that TZF and TZF-L may act to control the gene activity during spermatogenesis. PMID- 12589824 TI - Novel structural class of four disulfide-bridged peptides from Tityus serrulatus venom. AB - A new structural class of short peptides folded by four disulfide-bridges was found in the venom of the Brazilian scorpion Tityus serrulatus. Peptides were put on evidence independently by means of two different approaches of structurally guided prospection. First, a cDNA sequence was obtained using a degenerate primer constructed according to the C-terminal sequence of kaliotoxin (KTx2), from the Androctonus australis venom. Second, MALDI-TOF mass spectrometry analyses of toxic fraction FIII from T. serrulatus venom revealed a family of molecules ranging approximately from 2900 to 3000 Da. Three new peptides were isolated and named TsPep1, TsPep2, and TsPep3. Biochemical characterization showed that they are 29 amino acids long, constrained by a new pattern of four disulfide-bridges. These results enable us to classify these new molecules as part of a novel structural class of short peptides from scorpion venoms. PMID- 12589825 TI - General function of N-terminal propeptide on assisting protein folding and inhibiting catalytic activity based on observations with a chimeric thermolysin like protease. AB - Pro-aminopeptidase processing protease (PA protease) is a thermolysin-like metalloprotease produced by Aeromonas caviae T-64. The N-terminal propeptide acts as an intramolecular chaperone to assist the folding of PA protease and shows inhibitory activity toward its cognate mature enzyme. Moreover, the N-terminal propeptide strongly inhibits the autoprocessing of the C-terminal propeptide by forming a complex with the folded intermediate pro-PA protease containing the C terminal propeptide (MC). In order to investigate the structural determinants within the N-terminal propeptide that play a role in the folding, processing, and enzyme inhibition of PA protease, we constructed a chimeric pro-PA protease by replacing the N-terminal propeptide with that of vibriolysin, a homologue of PA protease. Our results indicated that, although the N-terminal propeptide of vibriolysin shares only 36% identity with that of PA protease, it assists the refolding of MC, inhibits the folded MC to process its C-terminal propeptide, and shows a stronger inhibitory activity toward the mature PA protease than that of PA protease. These results suggest that the N-terminal propeptide domains in these thermolysin-like proteases may have similar functions, in spite of their primary sequence diversity. In addition, the conserved regions in the N-terminal propeptides of PA protease and vibriolysin may be essential for the functions of the N-terminal propeptide. PMID- 12589826 TI - Two membrane proteins located in the Nag regulon of Candida albicans confer multidrug resistance. AB - Pathogenic fungus Candida albicans can efficiently utilize the aminosugar N acetylglucosamine (GlcNAc) as energy source. Since the mucosal membrane, the site of infection is rich in amino sugars, this specific adaptation is important for the establishment of infection. The genes encoding for the enzymes of the GlcNAc catabolic pathway, GlcNAc kinase (HXK1), GlcNAc-6-phosphate deacetylase (DAC1), and glucosamine-6-phosphate deaminase (NAG1), are present in a cluster, the Nag regulon, which is associated with virulence. In this study, we have characterized two genes, TMP1 and TMP2, present within the Nag regulon, upstream to DAC1. They encode two membrane associated sugar transporters of the major facilitator superfamily (MFS). The null mutant of TMP1 and TMP2 is able to grow in GlcNAc, implying that they are not involved in GlcNAc transport. However, it shows increased susceptibility to a number of unrelated antifungal compounds such as cycloheximide, 4-nitroquinoline-N-oxide, and 1-10 phenanthroline. Northern blot analysis revealed that TMP1 and TMP2 are upregulated in response to these drugs, suggesting that they function as multiple drug efflux pumps. PMID- 12589827 TI - Amphiregulin is a potent mitogen for the vascular smooth muscle cell line, A7r5. AB - The regulation of amphiregulin, an epidermal growth factor (EGF) family member, and its effect on vascular smooth muscle cells (VSMC) were examined. Amphiregulin mRNA was upregulated by amphiregulin itself as well as alpha-thrombin. Amphiregulin caused an approximate 3-fold increase in DNA synthesis. Its effect on growth was compared with those of other mitogens, and was found to be approximately 3.5-, 2.4-, and 1.0-fold greater than those of endothelin-I (ET-I), alpha-thrombin, and platelet-derived growth factor-AB (PDGF-AB), respectively. As evidenced by Western blot analysis, amphiregulin stimulated the phosphorylation of p42/p44-mitogen-activated protein kinase (MAPK), p38-MAPK, c-Jun NH2-terminal protein kinase (JNK), and Akt/protein kinase B (PKB), respectively. By statistical analysis, the amphiregulin-induced growth effect was significantly decreased by the MAP kinase/ extracellular regulated kinase kinase-1 (MEK-1) inhibitor PD98059, p38-MAPK inhibitor SB203580, and phosphatidylinositol 3-kinase (PI-3 kinase) inhibitor wortmannin, respectively, but was not decreased by JNK inhibitor SP600125. These results suggest that amphiregulin is the most potent mitogen of the mitogens tested, and its growth effect is mediated at least in part through the p42/p44-MAPK, p38-MAPK, and PI-3 kinase-Akt/PKB pathways in VSMC. PMID- 12589828 TI - Induction of SM-alpha-actin expression by mechanical strain in adult vascular smooth muscle cells is mediated through activation of JNK and p38 MAP kinase. AB - Mechanical forces have direct effects on the growth and differentiation of vascular smooth muscle. The goal of this study was to examine the effects of cyclic mechanical strain on expression of smooth muscle-alpha-actin (SM-alpha actin), a marker for the differentiated state of vascular smooth muscle, in cultured rat aortic smooth muscle cells (VSMC). Cells grown on dishes coated with either laminin or pronectin were subjected to mechanical strain and effects on expression of SM-alpha-actin were evaluated using the Flexercell Strain Unit. Application of mechanical strain to cells in full media increased SM-alpha-actin protein expression and promoter activity. This was not associated with any effect on growth. Mechanical strain increased activity of all three members of the MAP kinase family (ERKs, JNKs, and p38 MAP kinase), with similar kinetics. Inhibition of either JNKs or p38 MAP kinase blocked the strain-induced increase in SM-alpha actin promoter activity, and expression of constitutively active forms of JNK or MKK6, a p38 kinase, increased promoter activity. These studies indicate that in adult VSMC, mechanical strain leads to increased expression of smooth muscle markers, resulting in a more contractile phenotype. PMID- 12589829 TI - PKC regulates the delta2 glutamate receptor interaction with S-SCAM/MAGI-2 protein. AB - Inside cells, membrane proteins are localized at particular surface domains to perform their precise functions. Various kinds of PDZ domain proteins have been shown to play important roles in the intracellular trafficking and anchoring of membrane proteins. In this study, we show that delta2 glutamate receptor is interacting with S-SCAM/MAGI-2, a PDZ domain protein localized in the perinuclear region and postsynaptic sites of cerebellar Purkinje cells. The binding is regulated by PKC (protein kinase-C) mediated phosphorylation of the receptor with a unique repetitive structure in S-SCAM/MAGI-2. Co-expression of both proteins resulted in drastic changes of the receptor localization in COS7 cells. These results show a novel regulatory mechanism for the binding of PDZ domain proteins and suggest that the interaction between delta2 receptor and S-SCAM/MAGI-2 may be important for intracellular trafficking of the receptor. PMID- 12589830 TI - Retraction of "The design, synthesis, and biochemical evaluation of derivatives of biphenyl sulfamate-based compounds as novel inhibitors of estrone sulfatase" [Biochem. Biophys. Res. Commun. 294 (2002) 180-183]. PMID- 12589831 TI - Molecular analysis of cerebrospinal fluid in viral diseases of the central nervous system. AB - The use of nucleic acid (NA) amplification techniques has transformed the diagnosis of viral infections of the central nervous system (CNS). Because of their enhanced sensitivity, these methods enable detection of even low amounts of viral genomes in cerebrospinal fluid. Following more than 10 years of experience, the polymerase chain reaction or other NA-based amplification techniques are nowadays performed in most diagnostic laboratories and have become the test of choice for the diagnosis of several viral CNS infections, such as herpes encephalitis, enterovirus meningitis and other viral infections occurring in human immunodeficiency virus-infected persons. Furthermore, they have been useful to establish a viral etiology in neurological syndromes of dubious origin and to recognise unusual or poorly characterised CNS diseases. Quantitative methods have provided a valuable additional tool for clinical management of these diseases, whereas post-amplification techniques have enabled precise genome characterisation. Current efforts are aiming at further improvement of the diagnostic efficiency of molecular techniques, their speed and standardisation, and to reduce the costs. The most relevant NA amplification strategies and clinical applications of to date will be the object of this review. PMID- 12589832 TI - Herpes simplex virus resistance to antiviral drugs. AB - Herpes simplex virus (HSV) infections are efficiently treated with antiviral drugs such as acyclovir (ACV). However, resistance has been reported, mainly among immunocompromised patients (prevalence around 5%) and particularly allogeneic bone marrow transplant patients (prevalence reaching 30%). Resistance to ACV is associated with mutations on one of the two viral enzymes involved in the ACV mechanism of action: thymidine kinase (TK) and DNA polymerase. In 95% of the cases, ACV resistance is associated with a mutation in the TK gene as this enzyme is not essential for viral replication, unlike viral DNA polymerase, which is rarely involved in resistance. Strains resistant to ACV are almost always cross-resistant to other TK-dependent drugs such as penciclovir and famciclovir. Resistant infections can be managed by foscarnet or cidofovir but both are more toxic than ACV. These drugs also inhibit viral DNA polymerase but they are active on most ACV-resistant HSV as they do not depend on TK; nevertheless virus resistant to ACV because of a mutation in the DNA polymerase may be cross resistant to these molecules. Published data on genetic characterization of resistant clinical isolates point out hot spots in viral TK and DNA polymerase genes. TK mutations associated with resistance are either insertion or deletion (codons 92 and 146 of TK gene) or substitution (codon 176-177, 336 of TK gene). DNA polymerase mutations are mainly located in conserved sites of the enzyme. A high level of gene polymorphism has also been reported for these genes, especially for TK. These results are useful for the development of rapid genotypic assays for the detection of mutations associated with resistance. PMID- 12589833 TI - Detection of herpes simplex virus DNA in dried blood spots making a retrospective diagnosis possible. AB - BACKGROUND: Herpes simplex virus (HSV) infections in neonates are associated with life-threatening disease. Early diagnosis and treatment with antiviral therapy has decreased the morbidity, mortality and long-term sequelae in surviving children. The aim of the study was to investigate if herpes simplex virus DNA detection in dried blood spots on filter papers (Guthrie cards) sampled for screening of metabolic diseases may contribute to early diagnosis of neonatal HSV infection and enable pre-emptive therapy. METHODS: For detection of HSV-1 and -2 DNA, two different DNA extraction methods were evaluated. A minimal essential medium (MEM) extraction method was found superior and was used in combination with detection of HSV-1 and -2 DNA by PCR in dried blood spots from children with verified neonatal HSV infection. Cards from 28 children were included. The onset of illness varied from day 0 to 42 days and was the result of different types of maternal infection (27 cases) and an external source (one case). RESULTS: HSV DNA was detected in seven of the 28 Guthrie cards, two were HSV-1 and five were HSV-2 DNA positive. Positive dried blood spot cards were sampled within the interval 5 days before, to 6 days after onset of neonatal herpes. In cases of late onset CNS disease, viremia, was not demonstrable at the age of 3-5 days, the time period when the blood spot cards are normally sampled. CONCLUSION: Viremia, the prerequisite for demonstrating HSV DNA in dried blood spot cards preceded the onset of illness by up to 5 days and lasted at least up to 6 days thereafter. Analysis of HSV DNA in dried blood spot cards may be of value in the diagnostic arsenal for early onset of neonatal herpes and also have a role in the follow up of a child exposed at delivery. As the majority of the later onset neonatal herpes encephalitis cases are missed, a large-scale neonatal screening does not seem appropriate. PMID- 12589834 TI - Development and clinical application of a fully controlled quantitative PCR assay for cell-free cytomegalovirus in human plasma. AB - BACKGROUND: Cytomegalovirus (HCMV) disease continues to be a major problem in certain patient groups, including bone marrow transplant (BMT) recipients. The quantification of HCMV genome is clinically useful for the diagnosis of HCMV disease, for the virological surveillance of high-risk patients and for monitoring antiviral therapy. OBJECTIVES: To develop a novel, robust, and fully controlled PCR (qPCR) for the quantification of HCMV DNA in plasma samples and to demonstrate its clinical usefulness in the BMT setting. STUDY DESIGN: The newly developed HCMV qPCR employs cell culture-derived murine CMV as an internal control for both extraction and amplification. Following amplification using common primers, detection of both internal control and patient HCMV amplicons is by specific probes and a chemiluminescence microtitre plate system. Its performance was evaluated using the routine non-quantitative nested HCMV PCR on whole blood (NQPCR) and correlated with clinical events such as disease and antiviral therapy. RESULTS: A high level of concordance (85.1%) was found between the novel assay and the NQPCR, with the qPCR being slightly more sensitive. The samples giving discordant results generally had levels of HCMV DNA close to the limit of detectability or had been stored for prolonged periods. CONCLUSIONS: The use of plasma as an analyte by the newly developed assay avoids the detection of cell-associated virus. On the other hand, testing a comparatively large volume of plasma ensures that sensitivity is not compromised by not detecting cell associated HCMV. In a small preliminary evaluation in BMT recipients, changes in HCMV 'viral load' correlated with initiation and discontinuation of antiviral therapy and were biologically plausible. PMID- 12589835 TI - Distribution of the different genotypes of HCV among patients attending a tertiary care hospital in south India. AB - BACKGROUND: Genotyping of the hepatitis C virus (HCV) and assessment of viral load is important for designing therapeutic strategies and region specific diagnostic assays. OBJECTIVES: To determine the distribution of HCV genotypes among patients attending a tertiary care hospital in south India, and to correlate this with viral load. STUDY DESIGN: Ninety HCV RNA positive patients were recruited for the study. HCV genotyping was carried out using type-specific primers from the core region of the viral genome [J. Clin. Microbiol. 35 (1997) 201]. Viral load estimations were carried out using the Amplicor HCV Monitor (Versions 1.5 and 2, Roche Diagnostics, Branchburg, NJ, USA). Clinical details were elicited from patients' hospital records. RESULTS: Genotype 3 was detected most frequently (62.2%) followed by infection with HCV genotype 1 (18.8%). There was no significant difference seen in alanine aminotransferase (ALT) values between the two genotypes. Genotype 1 was associated with a significantly higher viral load as compared with genotype 3 (P=0.001). Parenteral transmission accounted for 61% of all infection caused. Infection with genotype 1 was significantly associated with a history of haemodialysis (P=0.01). Genotype 3 was detected more frequently in patients from east India, as compared with its detection in patients from south India (P=0.004). Similarly, genotype 1 was detected with greater frequency in individuals from south India as compared with patients from east India (P=0.004). The concordance between Ohno's genotyping assay and nucleotide sequencing, for genotypes 1 and 3, was 75%. CONCLUSIONS: HCV genotypes 1 and 3 accounted for 81% of HCV infections in patients from this geographical region. HCV genotype distribution showed regional differences and genotype 1 was associated with higher viral loads. Parenteral transmission was the major route for acquisition of HCV infection. Ohno's type-specific primer based genotyping assay can be used for distinguishing between HCV genotype 1 and non-1 HCV genotypes in laboratories that do not possess nucleotide sequencing facilities. PMID- 12589836 TI - BK virus (BKV) quantification in urine samples of bone marrow transplanted patients is helpful for diagnosis of hemorrhagic cystitis, although wide individual variations exist. AB - BACKGROUND: Hemorrhagic cystitis (HC) in allogeneic bone marrow transplanted (BMT) patients is associated with BK virus (BKV) reactivation manifested as BK viruria. However, since 77-90% of all adult BMT patients excrete BKV, viral reactivation alone cannot be responsible for HC. Recently, a significant overrepresentation of C-->G mutations in the Sp1 binding site in the non-coding control region (NCCR) of BKV was shown to be present in HC patients and absent in non-HC patients. OBJECTIVES: We aimed to investigate if this mutation resulted in excessive BKV excretion in HC patients. STUDY DESIGN: A Real-Time PCR was developed and used to quantify BKV in urine samples from 21 patients with HC, with and without the mutations, as well as from patients without HC. RESULTS: Quantification of BKV was successful in 18 of 21 urine patients (six with and six without C-->G mutations) and six patients without HC. A mean of 3.0 x 10(6) BKV copies/microl was detected in urine samples of HC patients with C-->G mutations, compared to a mean of 1.5 x 10(6) BKV copies/microl in HC patients without C-->G mutations and a mean of 1.0 x 10(6) BKV copies/microl in patients without HC. The obtained differences were however not statistically significant, due to one individual non-HC patient with an extremely high BKV copy number. Nevertheless, while 50% of the samples in the HC groups expressed 1 x 10(6) copies/microl or more, only one of the samples in the non-HC group contained a virus quantity higher than 5 x 10(5) copies. CONCLUSIONS: Although we could not confirm that the C-->G mutations in the Sp1 site of BKV were responsible for an increased viral load in patients with HC, our data suggest that levels of BKV above 10(4) copies/microl may indicate a risk for HC. PMID- 12589837 TI - Acute meningitis due to Toscana virus infection among patients from both the Spanish Mediterranean region and the region of Madrid. AB - Toscana virus (TOSV) is a member of the genus Phlebovirus that is transmitted to humans by two different species of sand fly and causes acute aseptic meningitis (AAM) and meningoencephalitis in Central Italy. Fifteen cases of AAM due to TOSV have been found at the Spanish province of Granada, but no data regarding the presence of TOSV-related disease in other regions of Spain have been still reported. A collection of 88 serum and 53 cerebrospinal fluid (CSF) samples taken from 81 selected patients with AAM of unknown aetiology, residing at Madrid or at the southern Mediterranean coast of Spain, was retrospectively studied for presence of TOSV-specific antibodies from both IgG and IgM classes. Anti-TOSV IgG was also investigated in 457 serum samples from healthy individuals, aged 2-60 years, residing at the south of the Region of Madrid. Specific IgM in serum and/or intrathecally produced anti-TOSV IgG were detected in seven patients, three residents from the Mediterranean region and the remainder four from the Region of Madrid. The overall prevalence of anti-TOSV among the healthy population studied was 5%. These results confirm the role of TOSV as an agent causing AAM in the Spanish Mediterranean coast, extend these findings to the central region of the country and suggest that TOSV might be producing infection and neurological disease in every area of Spain harbouring significant populations of the viral vectors. PMID- 12589838 TI - Parallel detection of five human herpes virus DNAs by a set of real-time polymerase chain reactions in a single run. AB - BACKGROUND: Human herpes viruses cause a spectrum of diseases that are usually self-limiting but can be reactive during immuno-suppression and may then lead to severe or even life-threatening diseases. The LightCycler technology allows rapid polymerase chain reaction (PCR) including product analysis within a closed system. This approach has been demonstrated to be suitable for routine diagnostic virus detection. Several LightCycler PCR assays have been established to the detection of human herpes viruses. The assays vary in their detection formats and PCR cycling protocols. So, they cannot be performed within a single LightCycler run. OBJECTIVES: Development of four LightCycler PCR assays for parallel detection of DNA derived from human cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), and varicella-zoster virus (VZV) in a single run. STUDY DESIGN: Primers and hybridization probes were tailored to suit one LightCycler PCR program. LightCycler PCRs were established, detection limits were determined, and clinical samples were evaluated. RESULTS: With quantified herpes virus type specific DNA spiked into cerebrospinal fluid, serum or EDTA plasma the detection limits were found either at 500 or 250 viral DNA copies per ml depending on the virus DNA specific PCR and on the specimen type used. The applicability of the new LightCycler assays for routine molecular testing was evaluated by testing 96 clinical samples. CONCLUSION: The developed set of LightCycler PCRs permits parallel detection of CMV, EBV, HSV-1, HSV-2, and VZV in a single LightCycler run. The new molecular assays can easily be used to the rapid, simple, and convenient detection of herpes virus DNA in cerebrospinal fluid, serum and EDTA plasma in the routine diagnostic laboratory. PMID- 12589839 TI - Multiple relapses of human cytomegalovirus retinitis during HAART in an AIDS patient with reconstitution of CD4+ T cell count in the absence of HCMV-specific CD4+ T cell response. AB - BACKGROUND: While in the past human cytomegalovirus (HCMV) represented the major viral opportunistic pathogen in patients with AIDS, incidence of HCMV disease in HIV-infected patients drastically dropped after introduction of highly active antiretroviral therapy (HAART). However, cases of HCMV disease in HIV-infected patients treated with HAART have been reported. OBJECTIVE: A 38-year-old HIV infected patient developed HCMV retinitis in May 1999 after reaching a nadir of 69 CD4(+) T cells/microl. HAART and anti-HCMV treatments with parenteral ganciclovir (GCV) were started, resulting in HIV viremia suppression, rise in CD4(+) T cell count to >300 cells/microl and recovery from retinitis. Notwithstanding the apparent immune reconstitution, every attempt to discontinue GCV maintenance treatment was followed by a relapse of retinal lesions. Thus, HCMV-specific CD4(+) cellular immune response was investigated. RESULTS: Lymphoproliferation assay and cytokine flow cytometry analysis were performed repeatedly from November 1999 showing absolute lack of HCMV specific CD4(+) T cell response, in the presence of an efficient lymphoprolipherative response against another pathogen (Candida) or a mitogen (Phytohemoagglutinin). CONCLUSION: In some patients, immune reconstitution after HAART may be only partial, since lack of pathogen-specific CD4(+) T cell response may persist even in the case of a significant rise in the absolute CD4(+) T cell count. This case suggests that immunologic assays investigating specific immune response against HCMV in HIV infected patients may be more useful than the CD4(+) T cell count alone in assessing immune function reconstitution after HAART and in deciding interruption of anti-HCMV secondary prophylaxis. PMID- 12589840 TI - Ethnicity and discordance in plasma HIV-1 RNA viral load and CD4+ lymphocyte count in a cohort of HIV-1-infected individuals. AB - BACKGROUND: Guidelines for commencing therapy for HIV infection have been based upon HIV-1 RNA and CD4 lymphocyte thresholds. The influence of confounding factors such as gender, ethnicity and co-infections is unproven. OBJECTIVES: To analyse ethnic discordance in plasma HIV-1 viral load (VL) and CD4+ count and its potential clinical significance in Black and Caucasian groups. STUDY DESIGN: Retrospective, cross-sectional, observational study of 537 antiretroviral nai;ve HIV-1-positive individuals attending two East London clinics. Baseline data were obtained from individuals who registered at the clinic from November 1996 to August 1999. An analysis was performed comparing ethnic differences in plasma HIV 1 VL, CD4+ count, CD8+ count, co-infections, CDC disease category, AIDS-defining illnesses and mode of transmission. RESULTS: Plasma HIV-1 VL was significantly lower in Blacks (4.5 copies/ml versus 4.7 copies/ml; P<0.05) despite lower baseline CD4+ counts and similar rates of disease progression to Caucasian groups. This association remained for patients with less advanced disease after stratification for CD4+ count (CD4+ 200-500, VL 4.5 copies/ml versus 4.7 copies/ml, P<0.01; CD4+ >500, VL 3.4 copies/ml versus 4.3 copies/ml, P<0.001) and disease category (non-AIDS, 4.4 copies/ml versus 4.7 copies/ml; P<0.005). On multivariate analysis, the association persisted following adjustment for gender, age, co-infections, CD4+ count and mode of transmission. CONCLUSIONS: These results suggest that plasma HIV-1 VL is discordantly low in Black compared with Caucasian groups stratified for CD4+ count, in this cohort of antiretroviral nai;ve HIV-1-positive individuals living in London. Although there are a number of possible explanations for this finding, it has considerable clinical relevance for the management of Black HIV-1-infected patients within UK, with significant implications for the decision about when to commence antiretroviral or immune based therapies. PMID- 12589841 TI - Evaluation of a commercial ELISA for detecting Norwalk-like virus antigen in faeces. AB - A commercially available enzyme immunoassay, the IDEIA Norwalk-like virus (NLV) enzyme linked immunosorbent assay (ELISA; Dako Cytomation, Ely, UK) for detecting NLV antigen in faecal samples and determining the NLV genogroup was evaluated. The performance of the ELISA was compared with that of electron microscopy and the reverse transcription polymerase chain reaction by testing a panel of faecal samples collected from patients involved in outbreaks of gastroenteritis. When compared with reverse transcription-polymerase chain reaction (RT-PCR), the ELISA had a sensitivity and specificity of 55.5 and 98.3%, respectively. This compares with a sensitivity and specificity for EM of 23.9 and 99.2%, respectively. The sensitivity and specificity of the ELISA for determining the aetiology of a Norwalk virus-like outbreak, based on two or more positive samples within an outbreak, were 52.2 and 100% when two samples were collected from an outbreak and 71.4 and 100% when six or more samples were collected. The ELISA correctly identified the NLV genogroups of viruses previously characterised by partial DNA sequencing. The ELISA is a suitable alternative to the preliminary screening by EM for investigating outbreaks of gastroenteritis. Outbreaks, negative by ELISA should be examined by RT-PCR in order to detect strains non-reactive in the assay and virus strains from representative ELISA positive outbreaks should be characterised fully to allow the genetic diversity of NLVs co-circulating in the population to be described. PMID- 12589842 TI - Effects of acyclovir on facial nerve paralysis induced by herpes simplex virus type 1 in mice. AB - OBJECTIVES: Bell's palsy has recently been claimed to be caused by herpes simplex virus type 1 (HSV-1) infection. The anti-viral agent acyclovir is a specific inhibitor of herpesvirus replication, and the most effective agent for the treatment herpesvirus infection. The purpose of this experiment was to assess the effect of acyclovir on the facial nerve paralysis included by HSV-1 infection. METHODS: We succeeded in producing an animal model of acute and transient facial nerve paralysis induced with HSV-1 neuritis simulating human Bell's palsy. In this study, acyclovir administration was performed before and after facial nerve paralysis, and continued for 5 days. Controls were given phosphate-buffer saline (PBS) instead of acyclovir, and the incidence and duration of facial nerve paralysis was compared in the acyclovir groups and controls. RESULTS: The incidence of facial nerve paralysis was significantly lower in the group given acyclovir before the paralysis than in the controls, and the duration of facial nerve paralysis was shorter. CONCLUSIONS: Administration of acyclovir before the paralysis reduced the incidence and duration of facial nerve paralysis. Administration of acyclovir after the paralysis improved the duration of facial nerve paralysis. PMID- 12589843 TI - Factors affecting the results of nonsurgical treatment of secretory otitis media in adults. AB - OBJECTIVE: There is only limited knowledge of the factors influencing the results of nonsurgical treatment of secretory otitis media (SOM) in adults. The present study was carried out to determine the effect of the middle ear (ME) inflation and to clear up the factors affecting the results of conservative treatment of SOM. METHODS: A total of 198 adult patients were randomized to either a group receiving treatment with ME inflation for 10 days or to a group receiving ME inflation and antibiotics (oral Amoxicillin 500 mg 3/day) for 10 days or to a control group. A wide variety of anamnestic and clinical pre-treatment variables were assessed by administered questionnaires. Pneumo-otoscopy, tympanometry and pure-tone audiometry were performed before the treatment and at the 3-5th, 10th+/ 2 and 60th+/-5 day of the study. For the prognostic factors, univariate analyses were first performed to determine the significant predictors, which were afterwards entered into a stepwise logistic regression model. RESULTS: The effectiveness of treatment was 50.6% in Group A and 58.7% in Group B (control group-11%, P<0.001). The results of treatment were stable and were found unchanged 6 weeks after the treatment. The most important prognostic criteria were defined by analyzing the dynamics of tympanometry (OR, 2.17) and audiometry (OR, 10.27) during conservative treatment. The results of the treatment were related to the mastoid pneumatization, the pathology of paranasal sinuses, the previous history of SOM, the age, and the pre-treatment otoscopic data. CONCLUSIONS: The results suggest, that daily ME inflation can be effective treatment for adults patients with SOM having regard to the factors significant for the prognosis of treatment. PMID- 12589844 TI - Assessment of total middle ear reconstruction by patient survey correlated with clinical findings. AB - OBJECTIVE: Our objective was to assess, by patient survey and clinical examination, the results of correctional total middle ear reconstruction of problematic mastoid cavities following radical or modified radical mastoidectomy. SETTING: This study was performed in an academic tertiary referral center. PATIENTS: The study consisted of a retrospective survey, using the modified hearing satisfaction scale, of 50 patients, who had undergone total middle ear reconstruction of their problematic mastoid cavities. The survey results were correlated with their postoperative clinical findings. RESULTS: Postoperatively, 98% of these patients expressed satisfaction in terms of improvement of their preexisting otorrhea (median of 4.32 satisfied), 65% were satisfied with their hearing and 85% were satisfied with the overall of this operation. The survey results had a good correlation with the audiometric findings and the clinical findings, i.e. the surgery resulted in a dry ear in all 50 patients and 51% of these patients showed hearing improvement of more than 5 dBHL. CONCLUSIONS: This study indicates that most patients were satisfied with the outcomes of this operation. The patients' point of view and the postoperative clinical data both indicate that total middle ear reconstruction is an excellent procedure for correcting problematic mastoid cavities following a radical or modified radical mastoidectomy. The hearing satisfaction scale is a useful instrument for assessing patient satisfaction following this surgical procedure. PMID- 12589845 TI - The effects of the chorda tympani damage on submandibular glands: biometric changes. AB - OBJECTIVE: It was aimed to analyze the biometric changes in ipsilateral submandibular glands of patients with unilateral chorda tympani (ChT) section during otological operations, compared with change in size of the contralateral glands and with those of healthy subjects. METHODS: 29 patients with unilateral complete ChT section and 29 healthy subjects with identical ages, genders, and weights to the patient group were examined ultrasonographically. The patients having a mean duration to follow-up examination of 32 months (2-84 months) were subdivided into two groups by their time to follow-up as short-term patient group (2-12 months, 14 patients) and long-term patient group (13-84 months, 15 patients). The ultrasonographic dimensions and volumes of submandibular glands were compared statistically between the groups. RESULTS: In the patient group, the glands on the contralateral, non-operated side were found to be greater than the ipsilateral, denervated glands in terms of both paramandibular depth dimension (P<0.05) and volume (P<0.01). The differences could be determined only in long-term patient group. When comparing the submandibular glands of the patient group with those of the control group, it was found that paramandibular depth dimension and volume of the submandibular glands on the contralateral, non operated side were statistically greater (P<0.01). There was no difference between submandibular glands on the operated side of the patient group and those of the control group (P>0.05). CONCLUSION: The late (13-84 months) biometric results of ChT damage on submandibular gland were significant for increase in the size of the contralateral, non-denervated submandibular gland. An atrophying effect was not ascertained in the submandibular glands denervated parasympathetically due to the section of the ChT. PMID- 12589846 TI - Insertion of tympanic ventilation tubes as a treating modality for patients with Meniere's disease: a short- and long-term follow-up study in seven cases. AB - OBJECTIVES: As a treatment for patients with Meniere's disease, insertion of a ventilating tube in tympanic membrane was carried out, and the effect of the therapeutic modality was analysed. METHODS: Seven patients (four males and three females, age ranged 35-62 years) with active Meniere's disease were placed with ventilating tubes in the affected ear and postoperative change in symptoms, i.e. incapacitating vertigo and hearing loss were investigated. Disease severity was scored and evaluated pre- and postoperatively under the guidelines proposed by the Committee on Hearing and Equilibrium in the American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS, 1985). RESULTS: Of the seven cases, the number of patients and the degree in controlling vertigo after the treatment were: at 24 months (short-term) five cases with substantial, one with limited and one with insignificant control, then at 42 months (long- term) four cases with substantial, three with limited control, respectively. The degree of disability in patients at 24 and 42 months of the treatment was compatible with the level of vertigo controlling. On the other hand, patients' hearing level was not affected by the treatment. CONCLUSIONS: Exact explanation for the effect of ventilating tubes in tympanic membrane is vague and its therapeutic effect was limited. However, at least this treatment might represent a short-term effect for the reduction of persistent vertigo in some patients with Meniere's disease and so it might become a treatment option because of its simple and less-invasive procedure. PMID- 12589848 TI - Effects of visual input on galvanic body sway test of unilateral vestibular deficiency in patients with unilateral vestibular schwannoma. AB - OBJECTIVE: Galvanic body sway tests (GBSTs) are performed with eyes fixed and closed. However, quantitative effect of fixation on GBST has been unknown. The purpose of this study is to address this question. METHODS: We evaluated GBSTs of patients with unilateral vestibular schwannomas and normal controls, while their eyes were open and closed. We evaluated three GBST parameters: maximum amplitude of the response, velocity and latency of the onset of response. RESULTS: Closing the eyes diminished stability, resulting in increased amplitude and velocity of the responses. However, apparent contribution to the latency of response could not be found. Contribution of visual fixation seems to be greater in apparatus with vestibular hypo-function than without it. Unilateral weakness (UW) was calculated for each parameter, and significant correlation, between with eyes fixed and closed, was found only for velocity parameter. CONCLUSION: Fixation would stabilize the body of subjects, and diminish GBST responses. GBST with and without fixation can be evaluated together, using certain coefficient. On the other hand, fixation would not have any contribution to the latency of the GBST response. When we evaluate vestibular dysfunction of patients with unilateral vestibular schwannomas, velocity parameter should be employed. PMID- 12589847 TI - Compensatory changes in static and dynamic subjective visual vertical in patients following vestibular schwanoma surgery. AB - OBJECTIVE: Among patients with vestibular schwanoma (VS), vestibular function is nonhomogeneous, both before and after surgical removal of the VS. This paper reports investigations of neural changes, especially changes in the contribution of visual input to vestibular system integration, after VS surgery. METHODS: We examined 33 patients who underwent VS surgery via a middle fossa approach. Static and dynamic subjective visual vertical (SVV) was measured once after surgery and compared to those measured in control subjects. SVVs were assessed using a paradigm requiring the subject to manually adjust an image of a bar to the perceived vertical alignment. SVVs were measured when the background was stationary or rotating. RESULTS: In almost all patients, static SVV deviated toward the operated side. In VS subjects, the mean static SVV was 1.8+/-2.2 degrees; the amount of deviation in the dynamic SVV toward the operated side (11.7+/-8.3 degrees ) was significantly larger than that to the intact side (8.8+/-5.5 degrees ). In VS subjects, static SVV was correlated with dynamic SVV only in cases of bar adjustments toward the operated side (R=0.67, P<0.001), but not in cases of adjustments toward the intact, unoperated side. The axis of rotation was defined as the mean value of dynamic SVV for adjustments toward either side. There was only a weak correlation between the static SVV and the axis of rotation (R=0.31; P<0.05) in the control subjects. On the other hand, a more robust correlation between static SVV and axis of rotation was found (R=0.67, P<0.001) in VS subjects. There was no correlation between the static SVV and the deviation of dynamic SVV from static SVV for CCW and CW in control subjects. In contrast, there were significant correlations between static SVV and deviation of dynamic SVV from static SVV for adjustments made toward both operated (r=0.48, P<0.001) and intact sides (r=038, P<0.05). CONCLUSION: It is assumed that the amount of deviation in static SVV reflects the individual level of compensation. In addition, increased visual dependency evoked a symmetrical bias of the dynamic SVV from the measures at initial SVV assessment (i.e. static SVV or the center of tilt). As a result, we conclude that the contribution of visual inputs had changed after surgery, while at the same time, each patient used their static SVV as their reference point for orientation. PMID- 12589849 TI - The effect of recombinant interferon alpha treatment on hearing thresholds in patients with chronic viral hepatitis B. AB - OBJECTIVES: To investigate the possible ototoxic effects of recombinant interferon alpha (IFN alpha) on patients with hepatitis B. PATIENTS AND METHOD: Twenty-seven adult patients (14 male, 13 female) with chronic hepatitis B who were administered recombinant IFN alpha treatment were selected for this study. These patients were treated with a dose of 10 MU three times per week during 8-12 months (average follow-up period was 11.2 month). The mean hearing thresholds at seven frequencies (250-8000 Hz) were determined the day before therapy, after 1st, 7th and 21st day, and 1 month after termination of treatment. RESULTS: Hearing loss was detected in nine of 27 patients on the 7th day and the degree of hearing loss increased until 21st day of treatment. It was not exceeded 20 dB for any frequency with continued IFN therapy. The hearing loss completely recovered 1 month after the cessation of the IFN alpha. When hearing thresholds values of the day before therapy, 1st day and 1 month after termination of treatment were compared with values of 7th and 21st days, a statistically significant difference was detected (P<0.001). But there was no significant difference between the hearing thresholds values of the day before therapy, 1st day and 1 month after termination of the treatment (P>0.05). CONCLUSION: IFN alpha administration in patients with hepatitis B may cause mild reversible sensorineural hearing loss. PMID- 12589851 TI - Clinical application of the Harmonic Scalpel to allergic rhinitis. AB - OBJECTIVE: The Harmonic Scalpel (HS) (Johnson & Johnson K.K.) is a surgical tool that allows cutting and coagulation using ultrasonic mechanical vibrations. Its use in the field of otolaryngology has recently been attracting interest. In this study we applied the HS on the inferior nasal concha using a sword shaped blade ('KATANA'). METHODS: Subjects were 20 patients with intractable, perennial allergies that resisted conservative therapy. Subjective symptoms were evaluated as one of five stages, 'symptoms disappeared' 'markedly improved', 'improved', 'unchanged', and 'aggravated'. RESULTS: At the 12th week, 18 patients (90%) with nasal congestion, 16 (80%) with nasal discharge, and 15 (75%) with sneezing had shown at least 'markedly improved'. CONCLUSION: Based on the improvement in subjective symptoms, local and histopathological findings, the HS is an effective surgical modality for these cases. We believe the HS is a safe and useful tool for nasal surgery. PMID- 12589850 TI - Establishment of osteoblast culture from human ethmoidal sinus. AB - OBJECTIVE: Chronic sinusitis is characterized by persistent chronic inflammation of the sinus system and local expression and release of various cytokines, such as IL-1 beta, IL-4, IL-6, IL-8, TGF-beta and TNF-alpha are deeply involved in the pathogenesis of the disease. We hypothesized that not only the sinus mucosa containing cells, such as epithelial cells and fibroblasts, but also osteoblasts, of which sinus bone structure consists, may contribute to this inflammatory network. This study evaluates the development and establishment of an osteoblasts culture system derived from human ethmoidal sinus as an initial step toward verifying this hypothesis. METHODS: Ethmoidal sinus bone was obtained from patients at the time of sinus surgery for chronic sinusitis and outgrowth cell sheets were obtained according to the explant-outgrowth cell culture technique. In order to examine the specific characteristics of osteoblasts in the obtained cells, four major features of osteoblasts (collagen type I, osteocalcin synthesis, alkaline phosphatase activity and extracellular matrix mineralization ability) were investigated at the third passage of the culture and productions of TGF-beta 1, which modulate osteoblast proliferation and maturation in an autocrine fashion, in the cultured medium was also investigated in time of culture up to 20 days. RESULTS: The cells obtained in our study clearly show collagen type I synthesis, osteocalcin synthesis, alkaline phosphatase activity and production of visible extracellular matrix mineralization. Production of TGF beta 1 in the medium did not significantly different in time of culture up to 20 days. CONCLUSION: Our results, the first of their kind, indicate that osteoblast like cells can be cultured from adult human ethmoidal sinus bones. It suggested that proliferation and maturation of the osteoblast were continuously modulated in an autocrine fashion by producing TGF-beta 1. PMID- 12589852 TI - KTP laser inferior turbinoplasty: an alternative procedure to treat the nasal obstruction. AB - OBJECTIVE: Nasal obstruction resulting from inferior turbinate hypertrophy (ITH) was treated with KTP laser inferior turbinoplasty (KIT). The effectiveness of the procedure was assessed. METHODS: A prospective clinical trial was carried out in King Chulalongkorn Memorial Hospital from October 1, 1998 to September 30, 2000. Forty-eight patients with chronic nasal obstruction underwent KIT. Nasal obstruction was pre- and postoperatively assessed, based on 4-point scale, by the patient and investigator. The scores were compared by paired t-test. The correlation of assessment by the patient and investigator was also demonstrated by weighted kappa test. Pre- and postoperative rhinomanometric evaluations were performed in 29 patients and were compared with paired t-test. RESULTS: Significant reduction of nasal obstruction was obtained from assessment by the patient (P<0.000) and by the investigator (P<0.000). The symptoms of sneezing, itching and rhinorrhea were significantly reduced postoperatively (P<0.000). The cure and improvement rate of nasal obstruction were at 70.8 and 100% (assessed by the patient) and at 77.1 and 100% (assessed by the investigator) respectively, and they showed a moderate correlation (Kw=0.65). Rhinomanometrically, the total airway resistance decreased but of not statistic significance (P=0.219), however, the inspired nasal airflow at 150 Pa and the volume of nasal cavities were significantly increased (P<0.00 and P<0.001, respectively). CONCLUSION: KIP was shown to effectively reduce the symptom and sign of nasal obstruction as well as other nasal symptoms without any significant complications. It should be an alternative method in treating the patients with nasal obstruction resulting from hyperplastic inferior turbinate. PMID- 12589853 TI - Effects of topical chlorhexidine applied to the rabbit nasal mucosa. AB - OBJECTIVE: To search the effects of administration of various concentrations of a wide-spectrum antimicrobial agent, chlorhexidine, to the nasal mucosa. MATERIAL AND METHODS: About 0.20, 0.12, 0.06 and 0.03% concentrations of chlorhexidine gluconate were applied to the rabbit nasal mucosa as one puff twice a day throughout 5 days. Another group, treated with serum saline to the nose, behaved as the control group. On the fifth day following drug administration, specimens were taken from nasal mucosa of the rabbits and examined under light microscope. RESULTS: As a result of comparison between drug treated group and control group, with increasing drug concentrations progressively increased neutrophil infiltration in mucosa, ciliary loss in cells, and occasional metaplasia were observed. CONCLUSION: There is a linear, positive and strong association between concentrations of chlorhexidine and its irritative effects on rabbit nasal mucosa. While 0.20 and 0.12% concentrations of chlorhexidine cause excess irritation on the nasal cavity, 0.06 and 0.03% concentrations of chlorhexidine gluconate causes lower irritation and effects on the animals which have experimentally induced rhinosinusitis must be evaluated. PMID- 12589854 TI - Improved outcomes with early vocal fold medialization for vocal fold paralysis after thoracic surgery. AB - OBJECTIVE: To determine the clinical impact of early versus late surgical therapy for new onset unilateral vocal cord paralysis (UVCP) after thoracic surgery. STUDY DESIGN: Patients diagnosed with new onset UVCP after esophagectomy, pneumonectomy or pulmonary lobectomy were reviewed to determine the incidence of pneumonia, need for postoperative bronchoscopy and length of stay (LOS). Comparisons were made between patients undergoing early (< or = 4 days after thoracic procedure) versus late rehabilitation (> or = 5 days after thoracic procedure) of their UVCP with vocal cord medialization. RESULTS: Some 86 patients (27 esophagectomies, 43 pneumonectomies and 16 lobectomies) with new onset UVCP were examined. A total of 32 patients (37.2%) underwent early vocal cord medialization and 54 (62.8%) underwent late repair. The pneumonia rate for patients undergoing early vocal cord medialization (6.3%) was significantly lower than the rate for vocal cord medialization (37.0%, P=0.001, chi(2)). Early medialization patients required fewer postoperative bronchoscopies (mean number of bronchoscopies, 0.26) than late medialization patients (mean bronchoscopies, 0.94, P=0.013). The median LOS was significantly decreased for early versus late medialization patients in both pneumonectomy (reduction in LOS of 8 days) and lobectomy groups (reduction in LOS of 7 days). CONCLUSIONS: Early vocal cord medialization decreases the pneumonia rate, the requirement for postoperative bronchoscopies and the LOS for patients suffering from new onset UVCP after thoracic surgery. SIGNIFICANCE: Consideration should be given to early medialization for new onset UVCP when medically sound. PMID- 12589855 TI - Post tonsillectomy morbidity: following the introduction of single-use instruments. AB - OBJECTIVE: Following the introduction of single-use-instruments (SUI) for tonsillectomy in January 2001 in UK, the Medical Devices Agency (MDA) received reports of significant increase in secondary haemorrhage in some centres. This was believed to be related to electrosurgical (diathermy) forceps. The aim of this study is to compare post tonsillectomy morbidity before (year 2000) and after (year 2001) the introduction of SUI in a unit where bipolar dissection is the most popular method of dissection. METHOD: The case notes of all tonsillectomy patients between 20 April and 31 October for the years 2000 (145 patients) and 2001 (166 patients) were reviewed. The patients' age and sex, other concurrent operations, grade of surgeon, dissection and haemostasis methods, length of post operative hospital stay and the reason for prolonged stay (if more than one night) were noted. Readmitted cases were analysed to identify the cause and the management. RESULTS: There was no case of reactionary haemorrhage in both the non SUI (year 2000) and the SUI (year 2001) groups. For the non SUI versus SUI group, 6.2 versus 4.8% stayed more than one night post operatively; 4.8 versus 5.4% were readmitted for secondary haemorrhage with 1.4 versus 2.4% return to theatre to control bleeding, respectively. The average hospital stay for the readmitted cases were 2.1 versus 1.6 days and the average time lapse between tonsillectomy and secondary haemorrhage were 6.7 versus 6 days respectively. CONCLUSION: There was no significant difference in post tonsillectomy morbidity following the introduction of SUI. PMID- 12589856 TI - Langerhans cell related inflammatory reaction in laryngeal squamous cell carcinoma. AB - The major cells involved in cancer cell kill are the T lymphocytes. However, T cells need to be activated upon antigen presentation, which is mediated by the antigen presenting cells, one of which is the Langerhans cell (LC). The purpose of this study was to assess the significance of LC and inflammatory cell infiltration in the laryngeal squamous cell carcinoma (LSCC). Forty-five patients who were operated on for LSCC between 1992 and 1999 were included in the study. The clinical and histopathological features of the patients were reviewed. A semiquantitative estimation of the lymphocyte dominant inflammatory reaction within and around the tumor was performed. Anti S-100 antibodies were used for immunohistochemical detection of LCs. Horseradish peroxidase method was used. LCs were present in almost all of the specimens within and around the tumor tissue. The S-100 results did not associate with grade, T and N stages, tumor stage, laryngeal site of involvement and survival (P>0.05). The S-100 results significantly associated with inflammatory reaction in the tumor tissue (P<0.01). In conclusion, the LC related response is not important to inhibit regional metastasis by cancer cells. The LC is not a reliable tool to determine prognosis of the patients with LSCC in the clinical practice. PMID- 12589857 TI - Orthostatic tinnitus: an otological presentation of spontaneous intracranial hypotension. AB - Spontaneous intracranial hypotension (SIH) is a syndrome characterized by orthostatic headache, low cerebrospinal fluid (CSF) pressure without apparent causes for CSF loss, and diffuse pachymeningeal gadolinium enhancement on cranial MRI. Hearing is affected in some patients with SIH. A case of SIH with orthostatic tinnitus is reported. A 51-year-old woman developed a severe headache that was almost completely relieved on recumbency. Cranial MRI with gadolinium infusion showed diffuse enhancement of the dura mater. Radionuclide cisternography demonstrated CSF leaks at the upper and lower thoracic levels. Epidural blood patches at these leak sites alleviated the orthostatic headache, however, orthostatic tinnitus and muffled hearing persisted. Initial audiometry was unremarkable; repeat audiometry performed 6 weeks later demonstrated low frequency hearing loss in the right ear. Continuous epidural saline infusion for 3 consecutive days was performed; auditory symptoms disappeared 4 weeks thereafter. This case illustrates that orthostatic tinnitus represent intracranial hypotension even in the absence of orthostatic headache. MRI with gadolinium infusion should be carried out in the evaluation of patients with orthostatic tinnitus. PMID- 12589858 TI - Pulsatile tinnitus alleviated by contralateral neck compression: a case report. AB - A case of a 58-year-old man with right pulsatile tinnitus originating from a small dural arteriovenous fistula (DAVF) of the jugular bulb is described. The tinnitus was alleviated by contralateral neck compression. This unusual observation ruled out venous pulsatile tinnitus, although a temporal bone CT scan showed a high jugular bulb. The fistula was confirmed by angiographic study. The patient was treated by transarterial embolization of the fistula and remains free of symptoms 1 year after treatment. PMID- 12589859 TI - Association of chromium exposure with multiple primary cancers in the nasal cavity. AB - A 56-year-old man who had worked at a chromate factory for 13 years developed squamous cell carcinoma of the left nasal cavity 11 years after retirement. He received intra-arterial chemotherapy, followed by surgery. Two years later, an adenocarcinoma was identified in the same nasal cavity just above the previous surgical region. He underwent medial maxillectomy in combination with postoperative irradiation. He has been disease free for 5 years after the second surgery. Microsatellite markers were examined in the second tumor specimen as a possible factor for carcinogenesis; however, replication errors were not observed in any of four loci (D2S123, D3S1067, TP53, D18S474) tested. The present case seems to have resulted from long-term exposure to chromium and, to our knowledge, is the first reported case with multiple primary cancers in the nasal cavity associated with chromium exposure. PMID- 12589860 TI - A case of internal carotid artery aneurysm diagnosed on basis of massive nosebleed. AB - The authors report a rare case of a nontraumatic giant internal carotid aneurysm which ruptured into the nasal cavity, causing massive rhinorrhagia in a 64-year old Japanese woman. CT images of the head revealed an aneurysm having a diameter of about 5 cm with a neighboring area of erosion of the anterior base of the skull, and a swelling extending to inside the sphenoidal sinuses. Cerebal angiography resulted in a diagnosis of a giant aneurysm of the left internal carotid artery. Coil embolization and other surgical treatments were recommended, but the patient's family refused to consent and requested only conservative therapy. The natural course of the patient was thus followed, and the patient died after repeated episodes of aneurysm rupture and massive nosebleeds. It was concluded that curative therapy should be administered as soon possible upon diagnosis of this disease. PMID- 12589861 TI - The radiological management of oesophageal food bolus obstruction using a gas forming agent and barium. AB - Removal of an impacted food bolus in the lower oesophagus at rigid oesophagoscopy is challenging and carries a significant risk of perforation. We present two cases which were successfully disimpacted radiologically, in order to raise awareness among otolaryngologists of this therapeutic option. PMID- 12589862 TI - A case of bilateral cervical lymph node metastasis of thyroglossal duct cancer. AB - We reported a case of thyroglossal duct cancer, which metastasized to the bilateral cervical lymph nodes. The cervical lymph node metastases were detected in the upper portion of the neck. According to the results obtained in the preoperative fine needle aspiration cytology, the lymph node metastases were classified as group V and a diagnosis of papillary carcinoma was made. The preoperative studies with 123I revealed no concentration in the submittal tumor. In our surgical procedure, bilateral supraomohyoid neck dissection was added to excision of the tumor. CONCLUSION: In view of the fact that thyroglossal duct cancer metastasized bilaterally to cervical lymph nodes in the present case, we experienced an extremely rare case. PMID- 12589863 TI - Maxillary cancer in a child: a case report. AB - Cases with maxillary cancer are most frequently found in the 5th to the 7th decades of life, and then rarely occur in adolescence, even less so in a children. One does not expect to encounter this tumor in children, considering that most have a non-epithelial origin. However, we had a case of maxillary squamous cell carcinoma which appeared in a 12-year-old boy. He was initially considered to have sinusitis, and received conservative treatment for a while. After diagnosis, a combination therapy consisting of preoperative irradiation along intra-arterial administration of 5-FU and CBDCA was followed by total maxillectomy and primary reconstruction. Unfortunately about 2 months later, distant metastasis developed along with local recurrence, and the boy died about 1 year 6 months after the initial presentation. The clinical details of this rare case will be discussed in conjunction with some important considerations in dealing with this type of malignancy in a child. PMID- 12589864 TI - Gastric cancer epidemiology and risk factors. AB - We performed a detailed analysis of the epidemiology of gastric carcinoma, based upon a review of the literature in English. The analysis reveals many puzzling features. There has been a steady fall in the incidence of gastric carcinoma in most societies studied, but a more recent steady rise in the incidence of adenocarcinoma of the cardia and lower esophagus, largely confined to White males. Although the evidence for a major role for Helicobacter pylori (H. pylori) in the etiology of gastric corpus cancer is compelling; in Western society, it probably accounts for fewer than half the cases. The relative roles of dietary constituents such as salt and nitrites and the phenotyping of H. pylori in causation and the beneficial effects of a high fruit and vegetable diet and an affluent lifestyle, for all of which there is some evidence, are yet to be quantified. PMID- 12589865 TI - Does questionnaire structure influence response in postal surveys? AB - This study tested the effect of questionnaire structure on response, speed of return, and content of answers in a postal survey. All 259 patients aged 30-59 years who consulted with back pain at four UK general practices from March to June 2001 were randomly allocated to receive either a traditionally or chronologically structured self-completion questionnaire. The response was higher and the returns quicker (P =.05) for the chronologic questionnaire. There were no statistically significant differences in completion rates or scores on the SF-36, Chronic Pain Grade, Hospital Anxiety and Depression Scale, or Roland-Morris Disability Questionnaire between the two types of questionnaire, and test-retest reliability was high for all scales. Changing questionnaire structure to make questions chronologic does not substantially affect the answers given, but may make a questionnaire more acceptable and easier to complete and speed up returns. PMID- 12589866 TI - Major adverse outcomes after percutaneous transluminal coronary angioplasty: a clinical prediction rule. AB - In this study, we developed and internally validated a clinical model for predicting major adverse outcomes in patients undergoing percutaneous transluminal coronary angioplasty (PTCA) using a multi-institutional prospective cohort study involving all adult patients who underwent PTCA at 12 participating institutions from August 1993 to October 1995. A major adverse outcome, defined as death, renal failure, myocardial infarction, cardiac arrest, stroke, or coma, occurred in 3.3 and 3.2% of patients in the derivation and validation sets, respectively. Death occurred in 1.5% in both sets. Fourteen variables were independently correlated with major adverse outcomes. The rule, which stratifies PTCA patients into six levels of risk based on the severity score, showed excellent discrimination (receiver-operating characteristic curve area 0.82) and calibration (Hosmer-Lemeshow chi-square statistic P =.90) and performed well on internal validation. This rule allows accurate preprocedure stratification of PTCA candidates according to their risk of suffering a major adverse outcome. PMID- 12589867 TI - Developing a prognostic model in the presence of missing data: an ovarian cancer case study. AB - When developing prognostic models in medicine, covariate data are often missing and the standard response is to exclude those individuals whose data are incomplete from the analyses. This practice leads to a reduction in the statistical power, and may lead to biased results. We wished to develop a prognostic model for overall survival from 1,189 primary cases (842 deaths) of epithelial ovarian cancer. A complete case analysis restricted the sample size to 518 (380 deaths). After applying a multiple imputation (MI) framework we included three real values for each one imputed, and constructed a model composed of more statistically significant prognostic factors and with increased predictive ability. Missing values can be imputed in cases where the reason for the data being missing is known, particularly where it can be explained by available data. This will increase the power of an analysis and may produce models that are more statistically reliable and applicable within clinical practice. PMID- 12589868 TI - Age at diagnosis and the choice of survival analysis methods in cancer epidemiology. AB - A young age at diagnosis of cancer is often seen as an indicator of the aggressiveness of the tumor. However, empirical studies have shown conflicting results on the association between age at diagnosis and survival. There are two choices of time scale for a Cox regression model: time since diagnosis, and age. The regression analysis of relative survival rates is an alternative to the Cox model. Using breast cancer data from a population-based cancer registry, we illustrate the features of Cox models using the two time scales and compare them with the relative survival approach. Using a Cox model with time since diagnosis as the time scale, a younger age at diagnosis is associated with a lower mortality; using age as the time scale gives the opposite result. The relative survival approach agrees with the Cox model with age as the time scale. We maintain that a careful clarification of research purpose and a careful choice of methods are necessary. PMID- 12589869 TI - How useful are unpublished data from the Food and Drug Administration in meta analysis? AB - The goals of this systematic review and meta-analysis were to ascertain whether studies of nonsteroidal anti-inflammatory drugs (NSAIDs) summarized in the FDA reviews are ultimately published, to compare the methodologic and population characteristics of studies summarized in the FDA reviews with those reported in peer reviewed literature, and to compare the pooled relative risk of dyspepsia from NSAIDs in each data source. Summary measures of risk difference were calculated with a random effects model; meta-regression was used to assess the effect of study covariates. Among 37 studies described in the FDA reviews, one was published. Sample size, gender distribution, indication for drug use, and methodologic quality did not vary significantly between the published and FDA data. The pooled risk ratio for dyspepsia obtained using published data (1.21) or FDA data (1.07) did not differ significantly or practically. Data from FDA reviews may be a viable data source for systematic reviews and meta-analyses but only after being subjected to the same methodologic scrutiny as published data. PMID- 12589870 TI - Comparative responsiveness of generic and specific quality-of-life instruments. AB - We assessed the relative responsiveness of generic and specific quality of life instruments in 43 randomized controlled trials that compared head-to-head 31 generic and 84 specific instruments. Using weighted effect size as the metric of responsiveness, we assessed the impact of instrument type, disease category, and magnitude of underlying therapeutic effect on responsiveness, and assessed the responsiveness of specific instruments relative to the corresponding domains of generic measures. In studies with a nonzero therapeutic effect, specific instruments (mean = 0.57) were significantly more responsive than generic instruments (mean = 0.39, P =.01), and than the corresponding domains of generic instruments (mean = 0.40, P =.03). Studies with low, medium, and high therapeutic effects showed a corresponding gradation in responsiveness differences between specific and generic instruments. We conclude that, overall, specific instruments are more responsive than generic tools, and that investigators may come to misleading conclusions about relative instrument responsiveness if they include studies in which the magnitude of the underlying therapeutic effect is zero. PMID- 12589871 TI - A short psychosocial questionnaire for patients with Parkinson's disease: the SCOPA-PS. AB - The purpose of this study was to develop a short questionnaire for psychosocial functioning in patients with Parkinson's Disease (PD). The SCales for Outcomes in Parkinson's disease-PsychoSocial questionnaire (SCOPA-PS) was tested in a survey and compared with other instruments and with medical information. This survey was sent to 205 patents with idiopathic PD. Eighty-six percent of the questionnaires were returned. Cronbach's alpha was 0.83. Two-week test-retest reliability was 0.85 (intraclass correlation coefficient). Construct validity with other scales (Spearman's rho) was 0.82 for the Parkinson's Disease Questionnaire-39-item version (PDQ-39), 0.76 for the PDQ-8, 0.69 for the Hospital Anxiety and Depression Scale, -0.61 for the Euroqol, and -0.60 for a visual analogue scale evaluating Quality-of-Life. The summary index revealed a significant increase with increasing disease severity. The SCOPA-PS is a new, short psychosocial questionnaire for patients with PD with good clinimetric properties. PMID- 12589872 TI - Rasch scoring of outcomes of total hip replacement. AB - We examined whether there are advantages in terms of outcome assessment of using Rasch methods of scoring the 12-item Oxford Hip Score questionnaire over conventionally summed scores. Data were collected on patients receiving total hip replacement surgery. Three patient groups were created according to surgery type: primary, revision, and re-revision; two groups were created according to satisfaction with surgery: very satisfied and dissatisfied. Analyses were performed to test the relative precision (RP) of Rasch scoring versus conventionally summed scores in discriminating the groups experiencing different types of surgery and level of satisfaction. At the 1-year follow-up, RP ratios favored the Rasch scoring method in both tests of discrimination. Considerable gains in precision were achieved with Rasch scoring methods when groups were compared in a cross-sectional way. Alternative approaches to scoring questionnaires should be investigated to better assess comparisons over time. PMID- 12589873 TI - Glucose screening and the risk of complications in Type 2 diabetes mellitus. AB - It is unknown whether glucose screening for Type 2 diabetes mellitus (DM2) reduces the risk of diabetic complications. We conducted a case-control study using 303 cases with DM2 and at least one symptomatic microvascular diabetic complication, matched 1:1 to control subjects. All subjects' blood glucose tests for the decade before the first clinical suspicion of DM2 were categorized as screening or not based on the presence of symptoms suggestive of DM2. Approximately 90% of case subjects and control subjects had been screened for diabetes. After adjusting for multiple covariates in a logistic regression model, the odds ratio of developing a complication associated with screening was 0.87 (95% confidence interval 0.38-1.98), suggesting that screening may be associated with a modest reduction in the risk of certain diabetic complications. However, the confidence limits were wide and consistent with no true benefit. Further studies are needed to establish whether the small reduction we observed is genuine. PMID- 12589874 TI - Short-term, intermediate-term, and long-term mortality in patients hospitalized for stroke. AB - Cerebrovascular disease is the third leading cause of death and the primary cause of long-term disability in the United States. Although the risk factors for stroke have been well defined, less is known about stroke mortality over varying time periods within the same cohort of patients. The purpose of this study is to define rates of short-term, intermediate-term, and long-term stroke mortality among patients experiencing a first-ever hemorrhagic or ischemic stroke between 1994 and 1998. Patients were identified from the Patient Treatment Files of the Department of Veterans Affairs (VA). We included all patients who were discharged from a VA inpatient facility with a diagnosis of acute stroke. Patients were excluded from the study if they had an admission within the previous 5 years for stroke or hemiplegia. We obtained information on the patient's age, gender, and coexisting illnesses. Unadjusted and adjusted 30-day mortality rates were computed using Kaplan-Meier analyses and Cox proportional hazards regression models. The survival-dependent Cox proportional hazards regression models were run for 31-90 days and 91-365 days from the index admission date, for patients who had survived to the start of each of these time periods. Separate models were run for ischemic (n = 34,866 patients) and hemorrhagic (n = 5,442 patients) strokes. Unadjusted 30-day mortality was 8.2 and 20.5% for ischemic and hemorrhagic strokes, respectively. The adjusted 30-day mortality rate was 7.4 and 18.8% for ischemic and hemorrhagic strokes, respectively. For ischemic stroke, age 65 years and older was associated with an increased risk for short-term, intermediate-term, and long-term mortality, while chronic heart failure was associated with an increased risk for short-term and long-term mortality. For hemorrhagic stroke, age 75 years and older, malignancy, and chronic heart failure were associated with increased mortality during all three time periods. Thirty day mortality is over two times greater following hemorrhagic stroke vs. ischemic stroke. For patients who survive 30 days after an ischemic stroke, the risk factor that remains significantly associated with long-term mortality, which may be improved with appropriate process of care, is chronic heart failure. For patients with a hemorrhagic stroke, variables that remain significantly associated with increased short-term and long-term mortality include malignant neoplasm and chronic heart failure. Information on stroke mortality is important for patients, physicians, and researchers. In addition to stroke treatment, clinicians must be able to provide families of stroke victims with appropriate prognostic information. Further work is needed to assess the impact of actual care patterns, for the above identified risk factors, on stroke prognosis over varying time periods. PMID- 12589875 TI - Long-term survival and cause-specific mortality in patients with cirrhosis of the liver: a nationwide cohort study in Denmark. AB - Mortality from cirrhosis of the liver has been examined in few long-term follow up studies. In the Danish National Registry of Patients, 1982-1989, we identified a cohort of 10,154 patients with liver cirrhosis and divided them according to the etiology of their liver disease. Causes of death were identified in the Danish Death Registry, 1982-1993. We estimated relative survival and standardized mortality ratios by comparing with the mortality in the general population. The 10-year relative survival was worse in patients with alcoholic cirrhosis (34%) or nonspecified cirrhosis (32%) than in patients with primary biliary cirrhosis (58%) or chronic hepatitis (66%). The standardized mortality ratio for all causes of death combined was 12-fold increased, 5-fold excluding cirrhosis-related causes. Mortality in all disease categories was increased, even in those not traditionally related to cirrhosis. In conclusion, patients with cirrhosis of the liver face reduced life expectancy due to several causes of death. PMID- 12589876 TI - Plasma C-reactive protein levels and their relationship to anthropometric and lipid characteristics among children. AB - C-reactive protein (CRP), a nonspecific marker of inflammatory status, is associated with cardiovascular disease (CVD) risk factors and the late occurrence of heart disease in adults. However, few studies assess the plasma CRP levels in healthy children. The purpose of this study is to evaluate the relationship between plasma CRP levels and anthropometric and lipid characteristics among children in Taiwan. After a multi-stage sampling of 85 junior high schools in Taipei, we randomly selected 835 children (410 boys and 425 girls) aged 12 to 16 years. Anthropometric and lipid profiles, including total cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C), and lipoprotein (a) were measured. We also calculated low-density lipoprotein cholesterol levels and the total cholesterol-to-HDL-C ratio as shown on the atherosclerotic index. In both genders, plasma CRP levels were significantly positively correlated with anthropometrics measures and inversely correlated with HDL-C levels. After adjusting for age, cigarette smoking, alcohol consumption, heart rate, and puberty development, children in the fourth quartile CRP subgroups were heavier and had significantly higher body mass index (BMI) and lower HDL-C levels than children with nondetected CRP. In multivariate regression models, CRP was significantly negatively associated with HDL-C levels even after adjusting for BMI in both genders. In this study, anthropometrics measures, especially BMI, were positively associated with plasma CRP levels. Furthermore, elevated CRP levels were associated with adverse lipids profiles. These data suggest that elevated plasma CRP levels might be associated with CVD risk factors that may be related to the late development of CVD in some Taiwanese children. PMID- 12589877 TI - Conditional versus unconditional logistic regression in the medical literature. PMID- 12589878 TI - Quantification of lateral ventricular subdivisions in schizophrenia by high resolution three-dimensional magnetic resonance imaging. AB - In vivo brain imaging and postmortem investigations have demonstrated ventricular enlargement in the brains of schizophrenic patients. However, the extent of changes in the volume of discrete ventricle subdivisions has not been clearly established. We conducted high-resolution three-dimensional magnetic resonance imaging in 40 schizophrenic patients (20 males and 20 females) and 40 healthy volunteers (20 males and 20 females). The lateral ventricle in each hemisphere was divided into the anterior horn, body, posterior horn and temporal horn. The volumes of the hemispheres, four subdivisions of the lateral ventricles and the third ventricle were measured. Compared to the control subjects, the bilateral hemisphere volumes were significantly lower in the patients than in the control subjects. In the lateral ventricular subdivisions of the male patients, the most substantial volume increase was in the left temporal horn, and volume increases were also observed in the bilateral anterior horns and the right body. The male patients also had a significantly increased volume of the third ventricle. The female patients showed similar patterns with less statistical significance. Thus, the schizophrenia patients showed ventricular enlargement, particularly in the left temporal horn, being more severely affected in the male than in the female. PMID- 12589879 TI - A preliminary morphometric magnetic resonance imaging study of regional brain volumes in body dysmorphic disorder. AB - Morphometric magnetic resonance imaging (MRI) was used to compare regional brain volumes in eight women with body dysmorphic disorder (BDD) and eight healthy comparison subjects. The BDD group exhibited a relative leftward shift in caudate asymmetry and greater total white matter vs. the comparison group. Findings with respect to the caudate nucleus are consistent with both the conceptualization of BDD as an obsessive-compulsive spectrum disorder, and the 'striatal topography model' of obsessive-compulsive disorders. PMID- 12589880 TI - CSF monoamine metabolites and MRI brain volumes in alcohol dependence. AB - Correlations between cerebrospinal fluid (CSF) concentrations of monoamine metabolites (MAM) and brain structure have been described in schizophrenia, but not in alcoholism. To investigate the relationship between monoaminergic transmission and brain structure in alcoholism, the metabolites of dopamine (homovanillic acid, HVA), norepinephrenine (3-methoxy-4 hydroxyphenylethyleneglycol, MHPG) and serotonin (5-hydroxyindoleacetic acid, 5 HIAA) were measured in lumbar CSF in 54 alcohol-dependent patients and 20 healthy subjects. The volumes of the cerebrum, total grey and white matter, total and ventricular CSF, left and right hippocampus, and corpus callosum area were measured with MRI. MHPG and age were positively correlated in alcoholic women. The MAM concentrations were not significantly correlated with the MRI volumes in the subject categories. There were no differences in MAM across subjects defined by diagnosis and gender, age of onset of alcoholism or comorbidity of psychiatric disorders. Total CSF, cerebrum, and white and grey matter tissue volumes differed between patients and healthy subjects. The greatest difference was the white matter reduction in alcoholic women. In alcoholic women and men, monoaminergic neurotransmission measured by the CSF MAM HVA, MHPG, and 5-HIAA is not significantly correlated with the size of different brain structures. PMID- 12589881 TI - Neural correlates of clinical symptoms and cognitive dysfunctions in obsessive compulsive disorder. AB - Although results from neuropsychological and neuroimaging studies have postulated the involvement of the frontal lobe and the subcortical brain regions in the pathophysiology of obsessive-compulsive disorder (OCD), neuroimaging studies have provided little evidence that cognitive abnormalities in patients with OCD are related to dysfunctions in these areas. This study was designed to determine whether the clinical features and cognitive deficits of OCD might be taken to reflect frontal-subcortical dysfunction. Fourteen patients with OCD and 14 case matched normal subjects completed clinical and cognitive evaluation, including four sets of neuropsychological tests that assessed the executive functions and visual memory. Cerebral glucose metabolic rates were measured by using positron emission tomography (PET) with 18F-fluorodeoxyglucose. Behavioral and PET data were analyzed using statistical parametric mapping for group differences and behavioral-metabolic correlates. The right orbitofrontal cortex showed increased metabolic activity and the left parieto-occipital junction showed decreased metabolic activity in patients. Metabolism in the right hippocampus, the left putamen and the right parietal region was associated with the severity of obsessive-compulsive symptoms. Correlations between metabolic rates and neuropsychological test scores in the prefrontal cortex and the putamen occurred only in the patient group. These results suggest that patients with OCD have distinct features of brain metabolic activities for performing cognitive tasks as well as presenting obsessive-compulsive symptoms. In particular, the frontal subcortical circuits might mediate not only symptomatic expression but also cognitive expression in patients with OCD. PMID- 12589882 TI - Cerebral blood flow in depressed patients: a methodological comparison of statistical parametric mapping and region of interest analyses. AB - Functional brain imaging has assumed a leading role in neuropsychiatric research. However, findings reported for mental disorders often vary. Whether this reflects diversity in pathophysiology or heterogeneity of imaging techniques and data analytic procedures is still unknown. This study compares region of interest (ROI) and statistical parametric mapping (SPM) analyses of a Tc99m-HMPAO single photon emission computed tomography (SPECT) imaging study of 23 depressed and 21 control subjects. Reduced regional cerebral blood flow (rCBF) was demonstrated by both methods in the right parietal and occipital lobes, but additional regions were identified only on ROI analysis (left temporal) and only on SPM analysis (left parietal). To investigate the contribution of SPM spatial normalization to these discrepancies, further ROI analyses were performed, applying the original ROI templates to normalized images, and applying regions identified by SPM to the original images. This study demonstrated considerable overlap in findings of SPM and ROI analyses. Differences between these methods may be mostly related to subjective placement of ROIs in ROI analysis, and standardized warping inherent in normalization in SPM. Given the advantages and drawbacks of each procedure, the choice of methodology should be determined in accordance with the study design, and complementary use of both methods may be considered. PMID- 12589883 TI - Rapid visuomotor preparation in the human brain: a functional MRI study. AB - An important feature of human motor behaviour is anticipation and preparation. We report a functional magnetic resonance imaging study of the neuronal activation patterns in the human brain that are associated with the rapid visuomotor preparation of discrete finger responses. Our imaging results reveal a large scale distributed network of neural areas involved in fast visuomotor preparation, including specific areas in the frontal cortex (middle frontal gyrus, premotor and supplementary motor cortex), the parietal cortex (intra parietal sulcus, inferior and superior parietal lobe) and the basal ganglia. Our reaction time results demonstrate that it is easier to prepare two fingers on one hand than on two hands. This hand-advantage phenomenon was associated with relatively enhanced levels of activity in the basal ganglia and relatively reduced levels of activity in the parietal cortex. These findings provide direct evidence for differential activity in a distributed brain system associated with specific neuro-computational operations subserving fast visuomotor preparation. PMID- 12589884 TI - Event-related brain potentials during auditory and visual word recognition memory tasks. AB - Event-related brain potentials (ERPs) recorded during presentation of a series of words or pictures show enhanced positivity between 300 and 800 ms after presentation of repeated items. However, little attention has been directed to the characterization of this ERP recognition memory effect using auditory stimuli. The present study directly compared the ERP 'old/new effect' for words presented in the visual and auditory modalities. Nose-referenced ERPs were recorded from 30 electrode sites while participants (N=16) were engaged in visual and auditory continuous word recognition memory tasks. Spatially and temporally overlapping ERP components were identified and measured by covariance-based principal components analysis. The expected old/new effect was observed in both modalities, with a comparable time course peaking at 560 ms, but having a more anterior scalp topography for visual items. This suggests a common cognitive process (i.e. successful retrieval of information from memory) associated with separable neural generators in each modality. Despite this temporal synchronization, the old/new effect overlapped ERP components having distinct scalp topographies (N2) or peak latencies (P3) for each modality. The positive going old/new effect was preceded by an earlier negativity peaking at 370 ms that was greater across modalities for old than new words, likely reflecting semantic processing aspects of word recognition memory. A late (beyond 900 ms), broadly distributed negativity was also greater for old than new words, prolonged for auditory items, and may represent activity of a post-retrieval process. PMID- 12589885 TI - A functional MRI study of high-level cognition. I. The game of chess. AB - Chess is a game that involves many aspects of high level cognition and requires sophisticated problem solving skills. However, there is little understanding of the neural basis of chess cognition. This study employed functional magnetic resonance imaging (fMRI) to identify cortical areas that are active during the analysis of chess positions compared with a spatial task with matched visual stimuli. Bilateral activation was revealed in the superior frontal lobes, the parietal lobes, and occipital lobes. Some small areas of activation were observed unilaterally in the left hemisphere. The left hemisphere showed more activation than the right. Results are discussed in relation to a similar brain imaging study on the game Go. PMID- 12589886 TI - A functional MRI study of high-level cognition. II. The game of GO. AB - GO is a board game thought to be different from chess in many aspects, most significantly in that GO emphasizes global strategy more than local battle, a property very difficult for computer programs to emulate. To investigate the neural basis of GO, functional magnetic resonance imaging (fMRI) was used to measure brain activities of subjects engaged in playing GO. Enhanced activations were observed in many cortical areas, such as dorsal prefrontal, parietal, occipital, posterior temporal, and primary somatosensory and motor areas. Quantitative analysis indicated a modest degree of stronger activation in right parietal area than in left. This type of right hemisphere lateralization differs from the modest left hemisphere lateralization observed during chess playing. PMID- 12589887 TI - Syntax-related ERP-effects in Dutch. AB - In two studies subjects were required to read Dutch sentences that in some cases contained a syntactic violation, in other cases a semantic violation. All syntactic violations were word category violations. The design excluded differential contributions of expectancy to influence the syntactic violation effects. The syntactic violations elicited an Anterior Negativity between 300 and 500 ms. This negativity was bilateral and had a frontal distribution. Over posterior sites the same violations elicited a P600/SPS starting at about 600 ms. The semantic violations elicited an N400 effect. The topographic distribution of the AN was more frontal than the distribution of the classical N400 effect, indicating that the underlying generators of the AN and the N400 are, at least to a certain extent, non-overlapping. Experiment 2 partly replicated the design of Experiment 1, but with differences in rate of presentation and in the distribution of items over subjects, and without semantic violations. The word category violations resulted in the same effects as were observed in Experiment 1, showing that they were independent of some of the specific parameters of Experiment 1. The discussion presents a tentative account of the functional differences in the triggering conditions of the AN and the P600/SPS. PMID- 12589888 TI - Nicotinic mechanisms of memory: effects of acute local DHbetaE and MLA infusions in the basolateral amygdala. AB - Nicotine has been shown to improve working memory. The neural mechanisms underlying this effect are still being determined. The ventral hippocampus is critical for nicotinic effects on memory. Local ventral hippocampal infusions of either the nicotinic alpha7 nicotinic receptor antagonist methyllycaconitine (MLA) or the alpha4beta2 nicotinic receptor antagonist dihydro-beta-erythroidine (DHbetaE) caused working memory impairments, but no additive effects were seen. Other areas, such as the amygdala, also likely play important roles in nicotinic effects on memory. Amygdalar lesions cause memory impairment and there is a dense concentration of nicotinic receptors in the basolateral amygdala. The current study used local basolateral amygdalar infusions of the nicotinic antagonists MLA and DHbetaE to determine the involvement of alpha7 and alpha4beta2 nicotinic receptors in spatial working and reference memory. Rats (n=8) were trained in the 16-arm radial maze and were implanted with bilateral infusion cannulae into the basolateral amygdala. Acute infusions of MLA (6.75 micro g/side, P<0.0005) or DHbetaE (3.38 micro g/side, P<0.025) caused significant working memory impairments. When given together MLA and DHbetaE did not produce an additive effect. In fact, the 6.75 micro g/kg dose of DHbetaE produced a significant (P<0.0005) attenuation of the MLA-induced working memory impairment. Significant effects were not seen with reference memory or response latency. Nicotinic systems in the basolateral amygdala, as in the ventral hippocampus, are important for spatial working memory. In both the basolateral amygdala and the ventral hippocampus, MLA and DHbetaE individually caused working memory impairments. The lowest effective dose of DHbetaE was lower in the basolateral amygdala than in the ventral hippocampus. In both the basolateral amygdala and the ventral hippocampus, combined MLA and DHbetaE treatment did not produce additive working memory deficits. Unlike in the ventral hippocampus, the addition of DHbetaE to MLA in the basolateral amygdala significantly reduced the MLA-induced working memory deficit. PMID- 12589889 TI - Dichotic listening performance and frontal lobe function. AB - In the present paper we report on speech perception in left (n=16) and right (n=10) frontal lobe nonaphasic patients compared with healthy controls (n=26). The patients were inpatients of the University Clinic of Neurology, Innsbruck, Austria, who participated in a prospective neuropsychological study dealing with several aspects of lateralized functioning. The patients were tested with dichotic presentations of consonant-vowel syllables, which allows for specific probing of speech perception in the left and right hemisphere, respectively. The task was to report which syllable they heard best on each trial, emphasizing single answers on each trial. There were 36 dichotic trials with pairs of CV syllables made up of the six stop consonants /p/, /t/, /k/, /b/, /d/, /g/, all paired with the vowel /a/. There were always two different syllables presented on each trial, one in each ear. The results showed significant differences between the left and right lesioned patients in their performance on the dichotic listening test. While the right lesioned patients and the control subjects had a normal and expected right ear advantage, the left lesioned patients performed almost at random with regard to the right and left ear stimulus. The right lesioned patients and the control group showed a right ear advantage of the same magnitude, although the overall performance was somewhat impaired in the right lesioned group compared to the healthy control group. The left lesioned patients showed no ear advantage at all, and particularly their right ear scores were impaired compared to both the control group and the right lesioned patient group. The results are discussed in terms of the role played by the left frontal cortex in speech perception and language asymmetry. PMID- 12589890 TI - Probability effects on event-related potential correlates of recognition memory. AB - A common finding in event-related potential (ERP) studies of recognition memory is that recognised items elicit greater positivity at parietal electrode sites than new items (the 'left parietal old/new effect'). Parietal positivity (the P300 or P3b) is also elicited in detection tasks with no memory demands by items of low probability of occurrence and high 'target value'. It has been argued that correctly recognised items are typically associated with lower probability and higher target value than new items, raising the question of the extent to which the old/new effect receives a contribution from, or interacts with, P300 activity. The present study explored this issue by comparing ERPs associated with correctly classified old and new items across three different ratios of old to new items: 25:75, 50:50 and 75:25. The left parietal old/new effect was not influenced by this manipulation in the latency range in which it is conventionally measured. Probability did influence the parietal ERPs to correctly recognised items post-800 ms; the scalp distribution of this probability effect was however qualitatively distinct from that of the preceding old/new effect. The left parietal old/new effect appears to be a relatively pure reflection of episodic retrieval, uncontaminated by the non-mnemonic factors of probability and target value. PMID- 12589891 TI - Motor and parietal cortical areas both underlie kinaesthesia. AB - Tendon vibration has long been known to evoke perception of illusory movements through activation of muscle spindle primary endings. Few studies, however, have dealt with the cortical processes resulting in these kinaesthetic illusions. We conceived an fMRI experiment to investigate the cortical structures taking part in these illusory perceptions. Since muscle spindle afferents project onto different cortical areas involved in motor control it was necessary to discriminate between activation related to sensory processes and activation related to perceptual processes. To this end, we designed and compared different conditions. In two illusion conditions, covibration at different frequencies of the tendons of the right wrist flexor and extensor muscle groups evoked perception of slow or fast illusory movements. In a no illusion condition, covibration at the same frequency of the tendons of these antagonist muscle groups did not evoke a sensation of movement. Results showed activation of most cortical areas involved in sensorimotor control in both illusion conditions. However, in most areas, activation tended to be larger when the movement perceived was faster. In the no illusion condition, motor and premotor areas were little or not activated. Specific contrasts showed that perception of an illusory movement was specifically related to activation in the left premotor, sensorimotor, and parietal cortices as well as in bilateral supplementary motor and cingulate motor areas. We conclude that activation in motor as well as in parietal areas is necessary for a kinaesthetic sensation to arise. PMID- 12589892 TI - Left hemispheric dipole locations of the neuromagnetic mismatch negativity to frequency, intensity and duration deviants. AB - The aim of the current study was to differentiate the sources of neuromagnetic mismatch negativity (MMNm) to deviants of different features. For this purpose, the MMNm of twenty-one healthy subjects (seven males) were recorded left hemispherically. Subjects were stimulated monaurally in an oddball paradigm with standard tones of 1000 Hz and three different kinds of mismatch tones (frequency, duration and intensity deviants). Data analysis revealed mean MMNm dipole locations anterior, inferior and more medial than the N100m dipoles. The mean difference between the N100m and MMNm dipoles was in the range of up to 6 mm in one dimension. The dipole locations of all three kinds of deviants differed significantly from each other. The MMNm dipoles of both frequency and duration deviants were found to be significantly inferior to the corresponding source of intensity deviants, while the MMNm dipoles of duration and frequency deviants significantly differed in anterior-posterior direction. This differentiation between sources emphasizes the importance of feature analysis in MMN(m) generation. PMID- 12589893 TI - Functional comparison of primacy, middle and recency retrieval in human auditory short-term memory: an event-related fMRI study. AB - Primacy and recency effects refer to the better performance or shorter response time on the first and last items than the middle ones of a memory list. In order to investigate its neural basis in auditory short-term memory, event-related fMRI was used to measure brain activities when subject was recalling the first, the last, or the middle items. Recalling the middle item was associated with more extensive activation in the left parietal and visual cortex, basal ganglia, and dorsal cerebellum. Recalling items from different serial positions also resulted in different activation time courses in the bilateral primary auditory cortex, left prefrontal cortex and left premotor cortex. These data indicate that the auditory cortex may serve as a transient storage or the auditory input buffer, which seems to play an important role in the primacy and recency effects. PMID- 12589894 TI - Frequency of occurrence of the F wave in distal flexor muscles as a function of hypnotic susceptibility and hypnosis. AB - The aim of the experiment was to assess whether the membrane excitability of flexor cervical and/or lumbar motoneurons is related to hypnotic susceptibility (measured with the Stanford Hypnotic Susceptibility Scale, Form A) and hypnosis. During the experimental sessions, hypnotized subjects were given only suggestions of relaxation (neutral hypnosis) while awake subjects were given instructions to be quiet and relaxed (simple relaxation). F waves were recorded from the abductor digiti minimi and abductor hallucis of both sides after electrical stimulation of the ulnar or tibial nerve, respectively, and were used as an index of motoneuron membrane excitability in three groups of subjects: highly susceptible under neutral hypnosis, highly susceptible and non-susceptible during simple relaxation sessions. In lower limbs, there was no difference among the groups in the frequency of occurrence of F waves. In upper limbs, hypnosis selectively reduced F frequency of occurrence in flexor motoneurons on the right side. This reduction persisted after the end of hypnosis and also occurred during the last period of relaxation in highly susceptible non-hypnotized subjects. Results support the idea that hypnotic phenomena develop along a continuum in which some trait differences can be more easily revealed by the induction of hypnosis. PMID- 12589895 TI - Complexity of visual stimuli and non-linear EEG dynamics in humans. AB - The effects of stimulus complexity on the nonlinear electrical brain (EEG) dynamics were investigated in a sample of 24 healthy volunteers. Stimuli used were either a single mechanical low-friction pendulum with a periodic movement (temporal frequency about 1 Hz) or a double-pendulum with a chaotic movement, which were observed for 2-3 min in each case. The prediction that a more complex visual stimulus (double-pendulum) increases the dimensional complexity of brain activity as compared to a simple visual stimulus (single-pendulum), was confirmed by determination of pointwise correlation dimension. Further, there was a significant decrease of alpha power in the double-pendulum compared to a single pendulum condition. Moreover, a correlation analysis showed a positive correlation between EEG complexity and beta power over the whole cortex in the single- and, above all, in the double-pendulum condition, and also a positive correlation between dimensional complexity and alpha power in the double-pendulum condition only, particularly in the brain regions responsible for the 'bottom-up' sustained attention processes. PMID- 12589896 TI - Intersentential syntactic context effects on comprehension: the role of working memory. AB - The study investigated the influence of a syntactically biasing context sentence on the processing of syntactically complex object-first relative clauses and even more complex object-first complement clauses in readers with individual differences in working memory. Behavioral as well as brain responses (event related potentials, ERPs) were recorded from two groups of participants with either a high or a low working memory span. The behavioral data taken from a post sentence comprehension question task indicate an intersentential syntactic contextual influence on the comprehension of object-first relative clauses for low span readers, but not for high span readers. The on-line ERP measures taken at the disambiguating item revealed for the high span readers a main effect of structure (subject- versus object-first) i.e., a P600, independent of context. Low span readers, in contrast, did not show any P600 structure effect. Thus, the combined data indicate a differential context effect on sentence comprehension in low and high span readers. High span readers parse the incoming information independent of intersentential context, whereas low span readers use the intersentential context off-line. PMID- 12589898 TI - Clinical importance of inter-episode symptoms in patients with bipolar affective disorder. AB - A literature review was performed to determine whether inter-episode symptoms are of clinical importance in the management of bipolar disorder. Inter-episode symptoms are easy to miss but observational studies confirm that they are related to impaired function and reduced survival to full relapse. Randomised, controlled trials with lithium carbonate, and two studies exploring psychological treatments to recognise and treat prodromal symptoms of manic or depressive relapse, suggest some inter-episode symptoms are worth recognising because they are associated with reduced manic relapse and improved function. A classification is proposed to inform attempts at management of inter-episode symptoms in bipolar disorder patients, both clinically, and for future research. However, promotion of well being in bipolar disorder patients does not require all symptoms to be treated. In other patients, the presence of inter-episode symptoms may be a marker of resistance to treatment. Finally, the mechanism by which inter-episode symptoms might lead to relapse, or even lead directly to functional impairment, awaits convincing explanation and empirical support. PMID- 12589899 TI - Relevance of new and newly rediscovered anticonvulsants for atypical forms of bipolar disorder. AB - The so-called atypical forms of bipolar disorder are not a rarity, but instead are rather the rule. Particularly in specialized settings such as the bipolar disorder clinic, the majority of patients are characterized by atypical manifestations (). Mixed states, psychotic mania and a rapid cycling course of bipolar disorder are a challenge both to pharmacological and non-pharmacological treatment. The benefit of classical mood stabilizers such as lithium and carbamazepine is limited in monotherapy, although valproate has a broader spectrum of activity in atypical bipolar disorders and is often used in combination with other agents. Thus, new treatment alternatives are needed urgently for optimizing the treatment of atypical bipolar disorder. During the last decade, several new antiepileptic drugs have been released, e.g. lamotrigine, gabapentin, tiagabine, topiramate and levetiracetam. Others have been available for some time, but only recently have become the focus of bipolar disorder research; for example, phenytoin, and especially, oxcarbazepine. This review will consider our current knowledge of the benefit of these new and newly rediscovered anticonvulsants in treating bipolar disorders, with a special focus on their value in treating atypical manifestations. PMID- 12589900 TI - Oxcarbazepine (Trileptal) in the treatment of bipolar disorders: a review of efficacy and tolerability. AB - Oxcarbazepine, although structurally similar to carbamazepine, is metabolised very differently. As a consequence, oxcarbazepine's pharmacokinetic profile is comparatively less complex, and a more predictable, linear relationship between oxcarbazepine and blood levels is apparent. Furthermore, hepatic enzyme induction is considerably less with oxcarbazepine. Unlike carbamazepine, oxcarbazepine does not appear to induce its own metabolism, nor is it highly protein bound. These pharmacokinetic and metabolic characteristics raise the expectation that potential for drug interactions and side effects with oxycarbazepine will be less than that reported for carbamazepine. This review compares the pharmacokinetic and metabolic profiles of the two drugs and the available efficacy and safety data of carbamazepine and oxcarbazepine, in the treatment of bipolar disorder. PMID- 12589901 TI - Integration of synaptic and intrinsic dendritic currents in cat spinal motoneurons. AB - The results of recent studies designed to reveal some of the 'rules' governing the integration of synaptic and intrinsic dendritic currents in spinal motoneurons are reviewed. When two or more sources of synaptic input are activated concurrently, their combined postsynaptic effects on cat spinal motoneurons with 'passive dendrites' are generally equal to or slightly less than those predicted from the linear sum of their individual effects. However, for experimental preparations in which active conductances on motoneuron dendrites are enabled, instances of greater-than- or less-than linear summation can occur. Further, these studies demonstrate that the persistent inward currents that are generated by motoneuron dendrites provide an intrinsic source of excitatory drive that is larger than those associated with any of the individual synaptic input systems studied to date. Since these intrinsic depolarizing currents can be rapidly inactivated by a hyperpolarizing input, they are ideally suited to providing a major source of the alternating 'drive' to motoneurons during locomotion. PMID- 12589902 TI - Metabotropic glutamate receptors provide intrinsic modulation of the lamprey locomotor network. AB - Spinal networks generate the motor pattern underlying locomotion. These are subject to modulatory systems that influence their operation and thereby result in a flexible network organization. In this review, we have summarized the mechanisms by which the different metabotropic glutamate receptor subtypes fine tune the cellular and synaptic properties and thus underlie intrinsic modulation of the activity of the locomotor network in the lamprey. PMID- 12589903 TI - Spinal interneurones; how can studies in animals contribute to the understanding of spinal interneuronal systems in man? AB - The first part of this review deals with arguments that the essential properties and organization of spinal interneuronal systems in the cat and in man are similar. The second part is concerned with the possibility that some interneuronal systems may be responsible for motor disturbances caused by spinal cord injuries and that these interneurones may be defined. This possibility is discussed with respect to the hyperexcitability of alpha-motoneurones and the exaggeration of stretch reflexes in spastic patients. To this end, what is known about cat spinal interneurones and about the neuronal basis and pharmacological treatment of spasticity, is put together. Interneurones in di- and trisynaptic reflex pathways from group II muscle afferents are singled out, since they are depressed by the alpha(2) noradrenaline receptor agonists clonidine and tizanidine, which is a critical feature of interneurones expected to contribute to exaggerated stretch reflexes which are reduced by alpha(2) noradrenaline receptor agonists. Recent observations that reflex excitation of extensor motoneurones from group II afferents is enhanced in spastic patients and that the pathologically strong reflex actions of group II afferents are reduced by clonidine and tizanidine support this proposal. On the other hand, a lack of effect of clonidine and tizanidine upon other types of excitatory or inhibitory interneurones argues against any major contribution of such interneurones to the abnormally strong responses of alpha-motoneurones to muscle stretch. PMID- 12589904 TI - The whole body receptive field of dorsal horn multireceptive neurones. AB - Multireceptive neurones are found in the spinal dorsal horn and may be projection neurones and/or interneurones for polysynaptic reflexes. The cutaneous receptive field of a multireceptive neurone exhibits a gradient of sensitivity with the centre responding to any mechanical stimulus, including hair movements and light touch, while the periphery responds only to noxious stimuli. These neurones also receive signals from viscera, muscles and joints. This convergence of inputs means that multireceptive neurones are continuously capturing all the information from both the interface with the external environment (the skin) and the internal milieu (the viscera, muscles, etc.). This information constitutes a 'basic somaesthetic activity' that could help the somatosensory system build a 'global representation of the body'. In addition to be seen as a global entity, the output of multireceptive neurones should be understood in dynamic terms since the size of the peripheral fields of the individual neurones may change, as a result of the plasticity of both excitatory and inhibitory segmental processes. Furthermore, the activity of these neurones can be inhibited from most of the remaining parts of the body via supraspinal mechanisms. These diffuse noxious inhibitory controls (DNIC) are triggered by peripheral A delta- and C-fibres, involve brain structures confined to the caudal-most part of the medulla including the subnucleus reticularis dorsalis (SRD) and are mediated by descending pathways in the dorsolateral funiculi. A painful focus that both activates a segmental subset of neurones and inhibits the remaining population can seriously disrupt this basic activity, resulting in the distortion of the body representation in favour of the painful focus, which becomes pre-eminent and (relatively) oversized. PMID- 12589905 TI - Activity of medullary serotonergic neurons in freely moving animals. AB - In the mammalian brain, serotonergic neurons in the medulla (n. raphe magnus, obscurus, and pallidus) send dense projections into the spinal cord, especially to the dorsal horn, intermediolateral column, and ventral horn. We have conducted a series of studies examining the single unit activity of these neurons in behaving cats. The experiments were directed at determining whether changes in unit activity were related to pain (n. raphe magnus), autonomic activity (n. raphe obscurus and pallidus), or motor activity (n. raphe obscurus and pallidus). The strongest relationship was between neuronal activity and motor output, especially tonic and repetitive motor activity. We hypothesize that the primary functions of this motor-related activity are to facilitate motor output, suppress processing of some forms of afferent activity, and to coordinate autonomic functioning with the current motor demand. PMID- 12589906 TI - Roles of primate spinal interneurons in preparation and execution of voluntary hand movement. AB - To study the contribution of primate cervical interneurons (INs) to preparation and execution of normal voluntary hand movement we investigated their activity and correlational linkages to muscles in monkeys performing tracking tasks. During ramp-and-hold flexion-extension torques about the wrist most task-related spinal INs exhibited some activity during both flexion and extension, in unexpected contrast to the strictly unidirectional activity of corticomotoneuronal (CM) cells and motoneurons. Most INs increased their activity more in one of these two directions; response patterns in their preferred direction were typically tonic or phasic-tonic. Spike-triggered averages of EMG detected significant features in muscle activity for many task-related INs. Premotor INs (PreM-INs) were identified by post-spike facilitation or suppression with appropriate onset latencies after the trigger spike. Muscle fields of PreM INs were smaller than those of supraspinal PreM cells in cortex and red nucleus, and rarely involved reciprocal effects on antagonist muscles. To investigate the relation of spinal INs to a repertoire of different muscle synergies, activity of INs was recorded from a macaque performing a multidirectional wrist task. The monkey generated isometric torques in flexion/extension, radial/ulnar deviation, pronation/supination, and executed a power grip that co-contracted wrist flexor and extensor muscles. Many INs showing task-modulated activity had preferred directions in this multidirectional space, typically with broadly tuned activation. The role of spinal INs in preparation for voluntary movement was revealed in monkeys performing instructed delay tasks. During the delay between a transient visual cue and a go signal a third of the tested INs showed significant delay modulation (SDM) of firing rate relative to the pre-cue rate. The SDM responses often differed from the INs' responses during the subsequent active torque period. In a monkey instructed by either visual or proprioceptive cues the delay period activity for many INs was similar in visual and perturbation trials, although other INs exhibited different SDM for visually and proprioceptively cued trials. These results suggest that spinal INs are involved, with cortex, in the earliest stages of movement preparation. The sensory input to INs could be identified in transient responses to the torque pulse, which showed two predominant patterns, consistent with inputs from cutaneous or proprioceptive receptors. We also investigated the task-dependent modulation of neural responses to peripheral input in a monkey performing wrist flexion/extension movements in a visually cued instructed delay task. Monosynaptic responses evoked by electrical stimulation of the superficial radial nerve through a cuff electrode were suppressed or abolished during the dynamic movement phase. Since task-related activity of the INs increased at the same time, the suppression was mediated by presynaptic rather than postsynaptic inhibition. These observations indicate that under normal behavioral conditions many spinal INs have response properties comparable to those previously documented for cortical neurons in behaving animals. PMID- 12589907 TI - Coordination and localization in spinal motor systems. AB - We review here experiments examining the hypothesis that vertebrate spinal motor systems produce movement through the flexible combination of a small number of units of motor output. Using a variety of preparations and techniques, these experiments provide evidence for such spinally generated units and for the localization of the networks responsible for producing them within different regions of the spinal cord. Such an organization might help to simplify the production of movement, reducing the degrees of freedom that need to be specified by providing a set of units involved in regulating features common to a range of behaviors. PMID- 12589908 TI - Modular organisation and spinal somatosensory imprinting. AB - The withdrawal reflex system has been extensively used as a model system for studies of pain related mechanisms, sensorimotor integration, learning and memory. For a long time, this system was assumed to be organised as a flexion reflex system. However, recent studies indicate that this system has a modular organisation, each module performing a detailed and functionally adapted sensorimotor transformation related to the withdrawal efficacy of its output muscle(s). Each module appears to be a self-organising circuitry that uses sensory feedback on single muscle contractions to adjust its synaptic organisation during development. These findings and their implications for the understanding of higher motor functions as well as clinical aspects will be discussed. PMID- 12589909 TI - Cellular bases of a vertebrate locomotor system-steering, intersegmental and segmental co-ordination and sensory control. AB - The isolated brainstem-spinal cord of the lamprey is used as an experimental model in the analysis of the cellular bases of vertebrate locomotor behaviour. In this article we review the neural mechanisms involved in the control of steering, intersegmental co-ordination, as well as the segmental burst generation and the sensory contribution to motor pattern generation. Within these four components of the control system for locomotion, we now have good knowledge of not only the neurones that take part and their synaptic interactions, but also the membrane properties of these neurones, including ion channel subtypes, and their contribution to motor pattern generation. PMID- 12589910 TI - Organization of left-right coordination in the mammalian locomotor network. AB - Neuronal circuits involved in left-right coordination are a fundamental feature of rhythmic locomotor movements. These circuits necessarily include commissural interneurons (CINs) that have axons crossing the midline of the spinal cord. The properties of CINs have been described in some detail in the spinal cords of a number of aquatic vertebrates including the Xenopus tadpole and the lamprey. However, their function in left-right coordination of limb movements in mammals is poorly understood. In this review we describe the present understanding of commissural pathways in the functioning of spinal cord central pattern generators (CPGs). The means by which reciprocal inhibition and integration of sensory information are maintained in swimming vertebrates is described, with similarities between the three basic populations of commissural interneurons highlighted. The subsequent section concentrates on recent evidence from mammalian limbed preparations and specifically the isolated spinal cord of the neonatal rat. Studies into the role of CPG elements during drug-induced locomotor like activity have afforded a better understanding of the location of commissural pathways, such that it is now possible, using whole cell patch clamp, to record from anatomically defined CINs located in the rhythm-generating region of the lumbar segments. Initial results would suggest that the firing pattern of these neurons shows a greater diversity than that previously described in swimming central pattern generators. Spinal CINs play an important role in the generation of locomotor output. Increased knowledge as to their function in producing locomotion is likely to provide valuable insights into the spinal networks required for postural control and walking. PMID- 12589911 TI - Development of posture and locomotion: an interplay of endogenously generated activities and neurotrophic actions by descending pathways. AB - The adult pattern of locomotion is observed at the end of the second postnatal week in the rat. The in vitro spinal cord isolated from immature rats has served as a valuable preparation to study the mechanisms underlying the development of locomotion. Although the rat is unable to walk at birth, because of an immature posture, its spinal cord networks can generate at least two kinds of motor patterns in vitro. One activity is called 'fictive locomotion' because it shares several common features with locomotion observed in vivo. This fictive locomotor pattern is rarely observed spontaneously and its release requires either pharmacological or electrical stimulation of the spinal cord. A second endogenously generated activity observed in this preparation occurs spontaneously and exhibits phase relationships between motor outputs that are quite different from the fictive locomotor pattern. Here we review some of the developmental functions this spontaneous activity may subserve. It is likely a major trigger for the maturation of lumbar networks in the fetus, at a stage when inputs from both the periphery and supraspinal structures are weak. Pathways descending from the brainstem arrive in the lumbar enlargement during the last week in utero and the first two postnatal weeks. These pathways, through the neurotransmitters they contain, especially monoamines, are essential for the expression of some neuronal properties and may regulate several ongoing developmental processes. PMID- 12589912 TI - Fast inhibitory synapses: targets for neuromodulation and development of vertebrate motor behaviour. AB - Locomotor networks must possess the inherent flexibility to adapt their output. In this review we discuss evidence from a simple vertebrate locomotor network that suggests fast inhibitory synapses are important targets for the forms of neuromodulation that afford this flexibility. Two important inhibitory transmitters, glycine and GABA, are present in the CNS of Xenopus tadpoles, where they each play distinct roles in the control of swimming. Glycine, but not GABA, contributes to the inhibitory mid-cycle component of each swim-cycle, the strength of which determines the frequency of swimming. Meanwhile, GABA release onto the swim network prematurely terminates swimming episodes. Hence, glycine controls how fast, whilst GABA controls how far the tadpole swims. Our work has focused on how the amines serotonin (5-HT) and noradrenaline (NA), and more recently the gas nitric oxide (NO), selectively target glycine and GABA release in the spinal cord to modulate swimming. In particular, we have identified three brainstem populations of nitrergic neurons, which suggests that nitric oxide may co-localise with 5-HT, NA and GABA. Here we review this work and suggest a hierarchy of brainstem modulatory systems, with NO acting as a metamodulator. PMID- 12589913 TI - Locomotor-like activity generated by the neonatal mouse spinal cord. AB - This report describes locomotor-like activity generated by the neonatal mouse spinal cord in vitro. We demonstrate that locomotor-like activity can be produced either spontaneously or by a train of stimuli applied to the dorsal roots or in the presence of bath-applied drugs. Calcium imaging of the motoneuron activity generated by a train of dorsal root stimuli revealed a rostrocaudally propagating component of the optical signal in the anterior lumbar (L1-L3) and in the caudal segments (S1-S4). We hypothesize that this spatio-temporal pattern arises from a rostrocaudal gradient of excitability in the relevant segments. Our experiments suggest that left/right reciprocal inhibition and NMDA-mediated oscillations are not essential mechanisms underlying rhythmogenesis in the neonatal mouse cord. Finally, our data are discussed in the context of other models of locomotion in lower and higher vertebrates. PMID- 12589914 TI - Spinal reflexes provide motor error signals to cerebellar modules--relevance for motor coordination. AB - The cerebellar olivo-cortico-nuclear network influencing rubro- and corticospinal tracts via the nucleus interpositus anterior (NIA) is one of the most thoroughly characterized mammalian motor systems involved in limb movement control. Recent findings indicate that climbing fibres innervating the NIA system mediate highly integrated sensorimotor information derived from spinal withdrawal reflex modules. In the present paper, the implications of this relationship between spinal and cerebellar neuronal networks for cerebellar sensorimotor processing are put in perspective of the modular organization of the NIA system. Data that should prove useful for computational models of cerebellar sensorimotor processing and motor learning, including functional spino-olivo-cortico-nucleo spinal connectivity, are reviewed. It is argued that spinal 'pre-processing' of climbing fibre input constitutes a signal conversion from 'sensory' to 'motor' coordinates, providing the cerebellar modules with motor error signals relevant to the action of single limb muscles. Drawing upon their patterns of interconnectivity with spinal reflex modules it is hypothesized how cerebellar modules may adaptively coordinate transitions between agonist and antagonist muscle activity. This mechanism would contribute to the generation of the triphasic EMG patterns that are necessary for smooth acceleration and deceleration of single-joint movements. PMID- 12589915 TI - Encoding and decoding of reticulospinal commands. AB - In the lamprey, the reticulospinal (RS) system is the main descending system transmitting commands to the spinal cord. We investigated these commands and their effect on the spinal mechanisms. The RS commands were studied by recording responses of RS neurons to sensory stimuli eliciting different motor behaviors. Initiation of locomotion was associated with symmetrical bilateral massive activation of RS neurons, whereas turns in different planes were associated with asymmetrical activation of corresponding neuronal groups. The sub-populations of RS neurons causing different motor behaviors partly overlap. We suggest that commands for initiation of locomotion and regulation of its vigour, encoded as the value of bilateral RS activity, are decoded in the spinal cord by integrating all RS signals arriving at the segmental locomotor networks. Commands for turns in different planes, encoded as an asymmetry in the activities of specific groups of RS neurons, are decoded by comparing the activities of those groups. This hypothesis was supported by the experiments on a neuro-mechanical model, where the difference between the activities in the left and right RS pathways was used to control a motor rotating the animal in the roll plane. Transformation of the descending commands into the motor responses was investigated by recording the effects of individual RS neurons on the motor output. Twenty patterns of influences have been found. This great diversity of the patterns allows the RS system to evoke body flexion in any plane. Since most neurons have asymmetrical projections we suggest that, for rectilinear swimming, RS neurons with opposite asymmetrical effects are co-activated. PMID- 12589916 TI - Contributions of the motor cortex to the control of the hindlimbs during locomotion in the cat. AB - Although the corticospinal tract is not essential for the production of the basic locomotor rhythm in cats, it does contribute to the regulation of locomotion, particularly in situations in which there is a requirement for precise control over paw placement or limb trajectory. Lesions of the dorsolateral funiculi at the low thoracic level (T(13)) that completely interrupted both the cortico- and rubrospinal pathways produced long-term deficits in locomotion on a level surface. These deficits included a paw-drag that was probably caused both by a loss of cortico- and rubrospinal input to motoneurones controlling distal muscles as well as by a change in the relative timing of muscles acting around the hip and knee. Smaller lesions produced similar deficits from which the cats recovered relatively quickly. Cats with the largest lesions of the dorsolateral funiculi were unable to modify their gait sufficiently to step over obstacles attached to the treadmill belt even 3-5 months postlesion. These results imply that the medial pathways, the reticulo- and vestibulospinal pathways, are unable to fully compensate for damage to the lateral pathways. Single unit recordings from identified pyramidal tract neurones (PTNs) within the hindlimb representation of the primary motor cortex (area 4) showed that a substantial proportion of neurones (67%) significantly increased their discharge frequency when the cats modified their gait to step over obstacles attached to the treadmill belt. Of those PTNs that showed increased activity during the swing phase, populations of neurones were activated at different times. A large proportion of PTNS discharged early in swing, in phase with knee flexors such as the semitendinosus. Others discharged slightly later, in phase with the activity of ankle flexors, such as tibialis anterior, while still others discharged at the end of swing, in phase with digit dorsiflexors, such as the extensor digitorum brevis. We suggest that different populations of cortical neurones may specifically modify the activity of selected groups of close synergistic muscles during different parts of the swing phase. We further suggest that these modifications are mediated, in part, by groups of interneurones that are involved in determining the base locomotor rhythm. This provides a means by which the changes specified by the descending signal from the motor cortex may be smoothly, and appropriately, incorporated into the locomotor cycle. PMID- 12589917 TI - Motoneuronal drive during human walking. AB - Recent technical advances have made it possible to reveal some of the inputs that drive spinal motoneurones during normal human walking. These techniques are based either on a temporary removal of the drive to the motoneurones or on an analysis of the coupling of motor unit activity. During walking a sudden unloading of the plantarflexor muscles leads to a pronounced drop in the soleus EMG activity. This unloading effect is caused by cessation of activity in the sensory afferents, which mediate positive feedback from the active muscles in the stance phase. Somewhat surprisingly the drop in EMG activity following unloading is still observed when Ia afferents are blocked, suggesting that these afferents do not make an important contribution to the motoneuronal drive. It would seem that gr. Ib and/or gr. II afferents are the main contributors to the positive feedback. It has been known for a long time that transcranial magnetic stimulation (TMS) at low intensities may selectively activate local inhibitory circuits in the cortex. At such low intensities TMS applied over the motor cortex may thus inhibit the output from the cortex. The removal of the corticospinal drive in this way may be revealed as a drop in EMG activity from the active muscle. During walking TMS may evoke such a drop in EMG activity from the active muscles, which demonstrates that the corticospinal tract makes a contribution to the muscle activity. Time- and frequency domain analysis of motor unit activity have been shown to be effective tools in the analysis of synaptic drive to spinal motoneurones during tonic voluntary contraction. Applying these techniques to human walking reveals that motor units recorded from the same muscle or from close synergists show short-term synchrony and coherence in the 15-20 Hz frequency band. However, motor units from muscles acting at different joints show no coupling. This suggests that leg muscles are generally activated relatively independently of each other during human walking. These techniques show great promises for revealing changes in the sensory and corticospinal drive to motoneurones in relation to different tasks as well as in patients after injury to the central motor system. PMID- 12589918 TI - Long-term potentiation in spinothalamic neurons. AB - Sensitization of nociceptive dorsal horn neurons, including spinothalamic tract (STT) cells, is thought to underlie the development of secondary hyperalgesia and allodynia following tissue injury. In central sensitization, responses to stimulation of sensory receptors are enhanced without any change in the excitability of the primary afferent neurons. We hypothesize that central sensitization of STT neurons is a variety of long-term potentiation (LTP). Evidence that LTP occurs in the spinal cord is reviewed. Neurotransmitters that trigger central sensitization include excitatory amino acids and peptides. Evidence for this is that co-activation of N-methyl-D-aspartate and NK1 receptors can produce long-lasting increases in the responses of STT cells, and antagonists of these receptors prevent central sensitization. Responses to excitatory amino acids increase and those to inhibitory amino acids decrease during central sensitization, presumably accounting for the changed excitability of STT cells. We believe these changes result from the activation of signal transduction pathways, including the protein kinase C, NO/protein kinase G and protein kinase A cascades. Recent evidence shows that calcium/calmodulin dependent kinase II (CaMKII) is also upregulated early in the process of central sensitization and that several types of ionotropic glutamate receptors become phosphorylated. It is proposed that the phosphorylation of neurotransmitter receptors leads to alterations in the sensitivity of these receptors and to central sensitization. Comparable events occur during LTP in brain structures. PMID- 12589919 TI - Role of kainate receptors in nociception. AB - Nociceptive nerve fibers use L-glutamate as a fast excitatory neurotransmitter and it is therefore not surprising that both, ionotropic and metabotropic glutamate receptors play pivotal roles for transmission of nociceptive information in spinal cord. A subtype of ionotropic glutamate receptors, the kainate receptor, is present in spinal dorsal horn. However, its role has remained obscure as specific antagonists and agonists have become available only recently. Kainate receptors are present on small, including nociceptive, dorsal root ganglion cells and on intrinsic dorsal horn neurons, and those two locations can be targeted separately by appropriate agonists and antagonists. Postsynaptic kainate receptors on spinal dorsal horn neurons are activated by high intensity electrical stimulation of the dorsal root entry zone that activates nociceptive primary afferent fibers. In contrast, low intensity stimulation that activates only non-nociceptive fibers is ineffective. Selective blockade of kainate receptors may produce analgesia. Here, we review what is known about localization of kainate receptors in dorsal root ganglia and spinal dorsal horn and their physiological and pathophysiological importance with special reference to nociceptive pathways. A short overview on molecular biology and agonist and antagonist pharmacology is included. PMID- 12589920 TI - Spinal plasticity mediated by postsynaptic L-type Ca2+ channels. AB - In the spinal cord, motoneurons and specific subgroups of interneurons express L type Ca(2+) channels. As elsewhere, these dihydropyridine-sensitive channels mediate a slowly activating inward current in response to depolarisation and show little or no inactivation. The slow kinetics for activation and deactivation provide voltage-sensitive properties in a time range from hundreds of milliseconds to tens of seconds and lead to plateau potentials, bistability and wind-up in neurons in both sensory and motor networks. This slow dynamics is in part due to facilitation of L-type Ca(2+) channels by depolarisation. The voltage sensitivity of L-type Ca(2+) channels is also regulated by a range of metabotropic transmitter receptors. Up-regulation is mediated by receptors for glutamate, acetylcholine, noradrenaline and serotonin in motoneurons and by receptors for glutamate and substance P in plateau-generating dorsal horn interneurons. In both cell types, L-type Ca(2+) channels are down-regulated by activation of GABA(B) receptors. In this way, metabotropic regulation in cells expressing L-type Ca(2+) channels provides mechanisms for flexible adjustment of excitability and of the contribution of plateau currents to the intrinsic properties. This type of regulation also steers the magnitude and compartmental distribution of Ca(2+) influx during depolarisation, thus providing a signal for local synaptic plasticity. PMID- 12589921 TI - Diversity of neurotrophin action in the postnatal spinal cord. AB - The expression of neurotrophins and their receptors in the adult spinal cord indicates that they have postnatal actions in addition to their well-known prenatal ones on axonal growth and cell survival. In this review we summarize evidence in support of mechanisms by which neurotrophins acutely modulate the response both of sensory neurons and of synapses within the spinal cord. The selective action of neurotrophins is achieved via restricted expression of high affinity trk receptors through which the neurotrophins act. Activation of trk receptors enhances the response of the vanilloid VR-1 receptor in nociceptive neurons leading to peripheral sensitization of the response to capsaicin or noxious heat. At synapses on motoneurons trk receptor activation enhances the response of NMDA receptors that in turn can increase the response of AMPA/kainate receptors on the same cell. Both of these sensitizing actions have a very rapid onset that is contrasted with slower neurotrophin effects on growth of axotomized afferents. It is likely that these different functional effects of neurotrophins reflect activation of different intracellular signaling pathways. These studies suggest mechanisms by which neurotrophins might be used to improve function of the damaged spinal cord. PMID- 12589922 TI - BDNF: a neuromodulator in nociceptive pathways? AB - During development, brain-derived neurotrophic factor (BDNF) supports the survival of certain neuronal population in central and peripheral nervous system. In adulthood, BDNF has been suggested to act as an important modulator of synaptic plasticity. This article reviews and discusses its potential role as neuromodulator in the spinal dorsal horn. BDNF is synthesized in the cell body of primary sensory neurons (pre-synaptic neurons) and its expression is regulated in models of inflammatory and neuropathic pain. The high affinity receptor for BDNF, tropomyosine receptor kinase B (TrkB), is expressed by post-synaptic neurons of the dorsal horn. Stimulation of pre-synaptic nociceptive afferent fibres induces BDNF release and consequent activation of TrkB receptors leading to a post synaptic excitability. Electrophysiological recordings showed that BDNF enhances the ventral root potential induced by C-fibre stimulation in an in vitro preparation. In addition, behavioural data indicate that antagonism of BDNF attenuates the second phase of hyperalgesia induced by formalin (in nerve growth factor-treated animals) and the thermal hyperalgesia induced by carageenan, suggesting that BDNF is involved in some aspects of central sensitisation in conditions of peripheral inflammation. In conclusion, BDNF meets many of the criteria necessary to define it as a neurotransmitter/neuromodulator in small diameter nociceptive neurons. PMID- 12589923 TI - Pain and rehabilitation after spinal cord injury: the case of sensory spasticity? AB - Sixty percent of patients with posttraumatic para- or tetraplegia suffer from severe, continuous burning and/or lancinating pain. Multiple sclerosis produces pain in more than 30%. This pain can be as important as the absent mobility or sexual function as a cause of lowered quality of life. Two unique types of longstanding neuropathic pain can be recognized in persons with spinal cord injury: (1). segmentally distributed pain at the lesion; and (2). pain in the body below the lesion, often with late onset. The first type could be produced by nerve root entrapment or by direct segmental deafferentation. The second type probably contains several forms of central pain, evoked either by the original spinal lesion, by an expanding syrinx in the spinal cord or by secondary changes at higher levels of the somatosensory systems. Patients with central pain almost always have stimulus-independent pain. Its intensity may vary independently, be related to the presence of visceral activity/inflammation or be constant. In addition, stimulus-dependent pain is sometimes present, usually because skin areas or viscera below the lesion are allodynic. Partial spinal lesions, especially centrally in the cervical spinal cord, may be more prone to produce pain than are complete lesions. There is limited analgesic effectiveness in controlled studies of serotonin reuptake inhibitors, of sodium channel blockers (lidocaine, tetracaine), of the GABA receptor agonist baclofen (one study) and of the NMDA-receptor antagonist ketamine (one study). There are anecdotal reports on oral carbamazepine, on gabapentin, on intrathecal opiates and also on the alpha(2)-agonist clonidine, being effective in central neuropathic pain. Neurostimulation is effective only if it evokes paraesthesia in the painful area; hence TENS may give relief of segmental pain. Neurodestructive procedures and central neurostimulation have been largely unsuccessful. As in other longstanding pain, improved coping through cognitive-behavioural rehabilitation may be helpful for the clinical outcome. PMID- 12589924 TI - Recovery of locomotion in the cat following spinal cord lesions. AB - In most species, locomotor function beneath the level of a spinal cord lesion can be restored even if the cord is completely transected. This suggests that there is, within the spinal cord, an autonomous network of neurons capable of generating a locomotor pattern independently of supraspinal inputs. Recent studies suggest that several physiological and neurochemical changes have to occur in the neuronal networks located caudally to the lesion to allow the expression of spinal locomotion. Some evidence of this plasticity will be addressed in this review. In addition, original data on the functional organisation of the lumbar spinal cord will also be presented. Recent works in our lab show that segmental responsiveness of the spinal cord of the cat to locally micro-injected drugs in different lumbar segments, in combination with complete lesions at various level of the spinal cord, suggest a rostro-caudal organisation of spinal locomotor control. Moreover, the integrity of midlumbar segments seems to be crucial for the expression of spinal locomotion. These data suggest that the regions of critical importance for locomotion can be confined to a restricted portion of the spinal cord. Later, these midlumbar segments could be targeted by electrical stimulation or grafts to improve recovery of function. Understanding the changes in spinal cord neurophysiology and neurochemistry after a lesion is of critical importance to the improvement of treatments for locomotor rehabilitation in spinal-cord-injured patients. PMID- 12589925 TI - Using robotics to teach the spinal cord to walk. AB - We have developed a robotic device (e.g. the rat stepper) that can be used to impose programmed forces on the hindlimbs of rats during stepping. In the present paper we describe initial experiments using this robotic device to determine the feasibility of robotically assisted locomotor training in complete spinally transected adult rats. The present results show that using the robots to increase the amount of load during stance by applying a downward force on the ankle improved lift during swing. The trajectory pattern during swing was also improved when the robot arms were programmed to move the ankle in a path that approximated the normal swing trajectory. These results suggest that critical elements for successful training of hindlimb stepping in spinal cord injured rats can be implemented rigorously and evaluated using the rat stepper. PMID- 12589926 TI - The effect of locomotor training combined with functional electrical stimulation in chronic spinal cord injured subjects: walking and reflex studies. AB - With the new developments in traumatology medicine, the majority of spinal cord injuries sustained are clinically incomplete and the proportion is likely to continue to rise. Thus, it is necessary to continue to develop new treatment and rehabilitation strategies and understand the factors that can enhance recovery of walking following spinal cord injury (SCI). One new development is the use of functional electrical stimulation (FES) device to assist locomotion. The objective of this review is to present findings from some recent studies on the effect of long-term locomotor training with FES in subjects with SCI. Promising results are shown in all outcome measures of walking, such as functional mobility, speed, spatio-temporal parameters, and the physiological cost of walking. Furthermore, the change in the walking behavior could be associated with plasticity in the CNS organization, as seen by the modification of the stretch reflex and changes in the corticospinal projection to muscles of the lower leg. In conclusion, recovery of walking is an increasing possibility for a large number of people with SCI. New modalities of treatment have become available for this population but most still need to be evaluated for their efficacy. This review has focused on FES assisted walking as a therapeutic modality in subjects with chronic SCI, but it is envisaged that the care and recovery of SCI in the early phase of recovery could also be improved. PMID- 12589927 TI - Transplantation of genetically modified cells contributes to repair and recovery from spinal injury. AB - The effects of transplantation of fibroblasts genetically modified to produce brain derived neurotrophin factor (Fb/BDNF) on rescue of axotomized neurons, axonal growth and recovery of function was tested in a lateral funiculus lesion model in adult rats. Operated control animals included those in which the lesion was filled with gelfoam implant (Hx) and those in which the cavity was filled with unmodified fibroblasts (Fb). Both Fb/BDNF and Fb transplants survived and filled the lesion site. Unoperated control groups showed a marked retrograde death of Red nucleus neurons contralateral to the lesion; Fb/BDNF recipients showed a significant rescue effect. Anterograde and retrograde labeling studies indicated no regeneration of rubrospinal axons into the lesion/transplant in operated control animals, but regeneration into, around, and through the transplant into the host was seen in the Fb/BDNF recipients. All animals showed deficits on the more challenging behavioral tests but the Fb/BDNF recipients showed fewer deficits, particularly in tests of spontaneous vertical exploration, horizontal rope crossing and a sensory test (patch removal). The improved function on these tests in the Fb/BDNF recipients was abolished by a second lateral funiculus lesion rostral to the transport site. These results indicate that delivery of neurotrophic factors by grafting genetically modified cells can improve repair and function after spinal injury. PMID- 12589928 TI - Strategies for repair of the deafferented spinal cord. AB - Deafferentation of the spinal cord by interruption of the sensory fibers in the dorsal roots highlights the problem of regeneration failure in the central nervous system. The injured dorsal root axons regenerate steadily, albeit slowly, in the peripheral compartment of the dorsal root, but abruptly cease to elongate when confronted with the interface between the peripheral and central nervous system, the dorsal root transitional zone (DRTZ). The glial cells of the CNS and their products together form this regeneration barrier. Recent years have witnessed several successful approaches to, at least in part, overcome this barrier. Particularly promising results have been obtained by (1). the replacement of adult non-regenerating dorsal root ganglion neurons with corresponding cells from embryonic or fetal donors, (2). the implantation of olfactory ensheathing cells at the DRTZ, and (3). immediate intrathecal infusion of growth factors to which dorsal root ganglion cells respond. In all these instances, growth of sensory axons into the adult spinal cord, as well as return of spinal cord connectivity, have been demonstrated. These findings suggest routes towards treatment strategies for plexus avulsion, and contribute to our understanding of possibilities to overcome regeneration failure in the spinal cord. PMID- 12589929 TI - Properties of motoneurons underlying their regenerative capacity after axon lesions in the ventral funiculus or at the surface of the spinal cord. AB - Spinal motoneurons represent neurons with axons located in both the central (CNS) and peripheral (PNS) nervous systems. Following a lesion to their axons in the PNS, motoneurons are able to regenerate. The regenerative capacity of these neurons is seen also after lesion in the ventral funiculus of the spinal cord, i.e. within the CNS compartment. Thus, after an axotomy within the ventral funiculus, motoneurons respond with a changing polarity towards production of axons, sometimes even from the dendritic tree. This capacity can be used in cases of ventral root avulsion (VRA) lesions, if a conduit for outgrowing axons is presented in the form of replanted ventral roots. In human cases, this procedure may accomplish return of function in denervated muscles. The strong regenerative capacity of motoneurons provides the basis for studies of the response in motoneurons with regard to their contents of substances related to survival and regeneration. Such studies have shown that, of the large number of receptors for neurotrophic substances and extracellular matrix molecules, mRNAs for receptors or receptor components for neurotrophin-3 (NT-3), ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF) are strongly downregulated after VRA, while receptors for glial cell line-derived neurotrophic factor (GDNF) and laminins are profoundly upregulated. These results should be considered in the design of combined pharmacological and surgical approaches to lesions of motor axons at or close to the CNS-PNS interface. PMID- 12589930 TI - Changes in mRNA content of developing opossum spinal cord at stages when regeneration can and cannot occur after injury. AB - The molecular mechanisms responsible for regeneration in the mammalian central nervous system (CNS) are poorly understood. Unlike the situation in adults, in the neonatal opossum, as in other immature mammals, the CNS shows successful regeneration after injury. We have used the isolated opossum CNS as a preparation for studying regeneration. An advantage of the opossum is that its developing spinal cord exhibits a gradient of regeneration in time and space. Thus, the potential for repair becomes lost in the cervical spinal cord when animals reach an age of 12 days or more. Animals up to 17 days of age still show regeneration in less mature lumbar segments of the spinal cord. To identify genes that underlie the process of regeneration we are studying mRNA changes in spinal cords at various stages of development. We have developed techniques for narrowing down the number of candidate genes by performing different gene subtraction experiments and by cross-hybridizing their results. This allowed us to select sequences differentially expressed in regeneration and to eliminate genes unrelated to that process. Our results reveal a number of novel sequences that could be important for spinal cord regeneration, as well as genes already supposed to play a role in regeneration. PMID- 12589931 TI - How do transplanted olfactory ensheathing cells restore function? AB - In this article, we review our work on regeneration of the corticospinal tract in rats following a lesion at upper cervical level. We outline the rationale for using olfactory ensheathing cells, and summarize the evidence for regeneration and functional recovery. The present interpretation on the mechanisms of functional recovery is partly hypothetical, and we emphasize where further experimental evidence is needed. PMID- 12589933 TI - Anabolic steroids, testosterone-precursors and virilizing androgens induce distinct activation profiles of androgen responsive promoter constructs. AB - Different androgens, e.g. virilizing androgens such as testosterone and its precursors as well as synthetic anabolic steroids, respectively, induce diverse biological effects. The molecular basis for this variety in biological actions, however, is not well understood. We hypothesized that this variability of actions may be due to steroid-specific target gene expression profiles following androgen receptor (AR)-activation. Therefore, we investigated androgen receptor dependent transactivation of three structurally different androgen responsive promoter constructs ((ARE)(2)TATA-luc, MMTV-luc, GRE-OCT-luc) in co-transfected Chinese hamster ovary (CHO)-cells as an artificial model simulating different natural target genes. Three virilizing androgens (dihydrotestosterone, testosterone, methyltrienolone), three anabolic steroids (oxandrolone, stanozolol, nandrolone) and two testosterone-precursors of gonadal and adrenal origin (dehydroepiandrosterone, androstenedione) were used as ligands (0.001-100 nM). All steroids proved to be potent activators of the AR. Remarkably, anabolic steroids and testosterone-precursors showed characteristic promoter activation profiles distinct from virilizing androgens with significantly lower (ARE)(2)TATA luc activation. Hierarchical clustering based on similarity of activation profiles lead to a dendrogram with two major branches: first virilizing androgens, and second anabolics/testosterone-precursors. We conclude that steroid specific differences in gene transcription profiles due to androgen receptor activation could contribute to differences in biological actions of androgens. PMID- 12589935 TI - Effect of 4-hydroxytamoxifen isomers on growth and ultrastructural aspects of normal human breast epithelial (HBE) cells in culture. AB - In the search for a breast cancer prevention strategy which would avoid undesirable effects of orally administered tamoxifen, the percutaneous administration of the highly active metabolite 4OHTamoxifen (4OHTam) has been proposed. Percutaneous 4OHTam penetrates the skin to reach breast tissues. It, thus, avoids the hepatic first pass effect, and offers an optimal local/systemic effect. However, trans-4OHTamoxifen can spontaneously isomerize into the cis isomer, which may have estrogen agonist action. The aim of this study was to examine the effect of cis-4OHTam on normal human breast epithelial (HBE) cells in culture. Spontaneous isomerization of trans- into cis-4OHTam occurred within 24 48h, but stabilized rapidly at a trans/cis ratio of 70/30, whether in stock solution, culture medium or cultured cells. The cis-4OHTam did not stimulate HBE cell growth according to histometric cell counts and scanning electron microscopy analysis, but inhibited E(2)-induced cell growth, albeit two to three times less than trans-4OHTam. In conclusion, spontaneous isomerization of trans- to cis-4 OHTam is limited and 4OHTam retains a marked antiestrogenic effect. It may prove to be a useful alternative to tamoxifen in breast cancer prevention, especially if administered percutaneously. PMID- 12589934 TI - Involvement of multidrug resistance proteins (MDR) in the modulation of glucocorticoid response. AB - Glucocorticoid resistance is a problem in the treatment of many diseases. One possible factor involved in the modulation of a glucocorticoid response is the export of glucocorticoids out of the cell. It has been shown that multidrug resistance protein 1 (MDR1, ABCB1), a member of the ABC family, is capable of transporting some glucocorticoids. This paper uses a mouse cell line, LMCAT in which the glucocorticoid response can be modulated by inhibitors of multidrug resistance proteins. Glucocorticoids fall into three categories. Firstly, those that are transported by an Abcb1a/Abcb1b transporter and whose transport can be inhibited by inhibitors of ABCB1 activity. Functional Abcb1a/Abcb1b was detected by inhibition of rhodamine efflux by these drugs and mRNA for Abcb1a and Abcb1b were detected in these cells. Secondly, those that are not transported. Finally, those that are transported by an Abcc1a transporter. Calcein transport out of these cells was blocked by treatment with probenecid indicating a functional Abcc1a transporter. Abcc1a mRNA was also detected in these cells. Thus, this paper provides insight into the mechanisms of glucocorticoid transport in cells and demonstrates a diversity of two independent mechanisms of transport of glucocorticoids by Abcb1a/Abcb1b and Abcc1a with individual patterns of steroid specificity. PMID- 12589936 TI - Alterations of expression and regulation of transforming growth factor beta in human cancer prostate cell lines. AB - TGF beta can promote and/or suppress prostate tumor growth through multiple and opposing actions. Alterations of its expression, secretion, regulation or of the sensitivity of target cells can lead to a favorable environment for tumor development. To gain a better insight in TGF beta function during cancer progression, we have used different cultured human prostate cells: preneoplastic PNT2 cells, the androgen-dependent LNCaP and the androgen-independent PC3 and DU145 prostate cancer cell lines. We have studied by specific ELISA assays in conditioned media (CM), the secretion of TGF beta 1 and TGF beta 2 in basal conditions and after hormonal treatment (DHT or E2) and the expression of TGF beta 1 mRNA by Northern blot. We have also compared the effect of fibroblast CM on TGF beta secretion by the different cell types. Compared to PNT2 cells, cancer cell lines secrete lower levels of active TGF beta which are not increased in the presence of fibroblast CM. LNCaP cells respond to androgen or estrogen treatment by a 10-fold increase of active TGF beta secretion while PC3 and DU145 are unresponsive. In conclusion, prostate cancer cell lines have lost part of their ability to secrete and activate TGF beta, and to regulate this secretion through stromal-epithelial interactions. Androgen-sensitive cancer cells may compensate this loss by hormonal regulation. PMID- 12589937 TI - Effects of 17 alpha-methyltestosterone on uterine morphology and heat shock protein expression are mediated through estrogen and androgen receptors. AB - Testosterone and the synthetic androgen, 17 alpha-methyltestosterone (MT), have been shown to increase uterine weights and alter uterine morphology. However, whereas the mechanism of action of testosterone in the uterus has been studied, it is not known if the actions of MT are mediated through androgen (AR) or estrogen (ER) receptors. In the present study, we have shown that MT, at 0.5 or 10 mg/kg per day, increases uterine weight and alters uterine morphology in a dose-dependent manner. Co-administration of the anti-androgen, flutamide, or the anti-estrogen, ICI 182,780, with MT revealed that the effects of the low dose of MT are mediated through the ER, whereas those of the high dose are mediated through both the ER and AR. In addition, we have studied the effects of MT on uterine heat shock proteins (hsps), a group of estrogen-regulated proteins whose levels increase in response to growth signals and protein damage. MT increased levels of hsp90 alpha, hsp72, and grp94. All effects on uterine hsp levels were antagonized by the anti-estrogen and not the anti-androgen. Collectively, the results of the present study indicate that the effects of MT in the uterus are mediated through the AR and ER. PMID- 12589938 TI - Estrogenic and antiproliferative activities of isoliquiritigenin in MCF7 breast cancer cells. AB - Licorice root contains chemically diverse compounds that exhibit estrogenic effects in vitro and in vivo. The chalcone isoliquiritigenin (ISL) is a component of licorice extract exhibiting either antitumorigenic activity or estrogen receptor (ER) alpha-dependent growth promoting effects on breast cancer cells. In order to contribute to a better understanding of this apparent paradox, we synthesized and ascertained the estrogenic properties of ISL using, as model systems, the hormone-sensitive MCF7 breast cancer cells and the steroid independent HeLa cells. Transfection experiments reveal that ISL is able to transactivate the endogenous ER alpha in MCF7 cells and this is supported by the capability to induce down-regulation of ER alpha protein levels and up-regulation of pS2 mRNA. Moreover, by using chimeric proteins consisting of the hormone binding domains of ER alpha and ER beta fused to the Gal4 DNA binding domain, we have determined that ISL is an estrogenic agonist of both ER isoforms. As a biological counterpart, low and intermediate ISL concentrations that induce substantial transcriptional activity stimulate the proliferation of MCF7 cells. However, high levels of ISL become cytotoxic even in steroid-receptor negative HeLa cells. Thus, the activity of ISL and the balance between risk or chemopreventive factor for estrogen-dependent breast cancer may depend on dietary intake. PMID- 12589939 TI - Real-time reverse transcription PCR assay of CYP19 expression: application to a well-defined series of post-menopausal breast carcinomas. AB - Aromatase, the product of the CYP19 gene, plays a key role in androgenic steroids transformation into estrogens from various hormonal sensitive tissues. Thus, in situ expression of CYP19 has been suggested to be involved in breast tumor growth especially in post-menopausal patients.We developed a real-time quantitative RT PCR assay based on fluorescent TaqMan methodology to quantify total CYP19 gene expression at the mRNA level in breast tumors. This method, based on nucleic acid quantification in homogeneous solutions, has the potential to become a standard in terms of its sensitivity, wide dynamic range and high-throughput capacity. In a well-defined series of 107 post-menopausal breast tumor samples, relative CYP19 mRNA levels ranged from 1 to 131. Among the four major CYP19 exon I-spliced transcripts, designated I.a, I.b, I.c and I.d, mRNA levels of the latter three correlated positively with total CYP19 mRNA levels. In ER alpha-positive breast tumors, CYP19 and ER alpha mRNA levels correlated negatively with each other (P=0.0078, r=-0.266), while CYP19 and ER beta mRNA levels correlated positively (P=0.00012, r=+0.388). Patients with high CYP19 mRNA levels did not relapse more frequently or have shorter relapse-free survival than other patients. Finally, mRNA levels of IL6, a major CYP19 regulatory factor, were significantly higher in tumors strongly expressing CYP19 than in tumors weakly expressing CYP19 (P=0.018). In conclusion, CYP19 expression did not influence the outcome of post menopausal patients with breast cancer. PMID- 12589940 TI - The intrinsic transcriptional estrogenic activity of a non-phenolic derivative of levonorgestrel is mediated via the estrogen receptor-alpha. AB - Levonorgestrel (LNG), a 19-nor-testosterone derivative, is widely used in contraceptive formulations. This compound does not bind to the estrogen receptor (ER), but it shows estrogen-like effects under in vivo and in vitro conditions. The estrogenicity of LNG may be attributed to its bio-transformation into non phenolic metabolites. In this study, the ability of A-ring reduced LNG metabolites to activate transcription via an estrogenic mechanism of action, including differences between ER alpha and ER beta subtypes, were investigated. Transactivation assays were performed in HeLa cells transfected with expression vectors for ER alpha and ER beta and an estrogen-responsive reporter gene. Cells were also transfected with expression vectors for both progesterone receptor (PR) isoforms (A or B). As expected, the tetrahydro derivatives of LNG (3 alpha,5 alpha- and 3 beta,5 alpha-LNG) showed significantly lower PR-mediated transcriptional activities through both isoforms when compared with progesterone (P(4)) and LNG. In contrast, the 3 beta,5 alpha-tetrahydro derivative resulted in a significant activation of estrogen-dependent gene transcription. This effect was selectively confined to the ER alpha, since little if any activity could be observed with the ER beta and no antagonistic activities were demonstrated. This study provides structural and molecular clues for the well documented in vitro and in vivo intrinsic estrogenicity of 19-nor-testosterone-derived progestins and ligand requirements for ER alpha recognition. PMID- 12589941 TI - Protection by estrogens of biological damage by 2,2'-azobis(2-amidinopropane) dihydrochloride. AB - We examined by using 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) as a radical generator the ability of estrogens to scavenge carbon-centered and peroxyl radicals. Electron spin resonance signals of carbon-centered radicals from AAPH were diminished by catecholestrogens but not by phenolic estrogens, showing that catecholestrogens efficiently scavenged carbon-centered radicals. However, fluorescent decomposition of R-phycoerythrin by AAPH-derived peroxyl radicals was inhibited by catecholestrogens and phenolic estrogens. Evidently, peroxyl radicals were scavenged by catecholestrogens and by phenolic estrogens. However, the scavenging ability of 4-hydroxyestradiol was less than 2 hydroxyestradiol. Strand break of DNA induced by AAPH was inhibited by catecholestrogens, but not by phenolic estrogens under aerobic and anaerobic conditions. Inactivation of lysozyme induced by AAPH was completely blocked by 2 hydroxyestradiol under aerobic and anaerobic conditions, and by 4-hyroxyestradiol only under anaerobic conditions. Peroxidation of arachidonic acid by AAPH was strongly inhibited by catecholestrogens at low concentrations. Only large amounts of phenolic estrogens markedly inhibited lipid peroxidation. These results show that catecholestrogens were antioxidant against AAPH-induced damage to biological molecules through scavenging both carbon-centered and peroxyl radicals, but phenolic estrogens partially inhibited AAPH-induced damage because they scavenged only peroxyl radicals. PMID- 12589942 TI - Temporal effect of tamoxifen on cytochrome P450 side chain cleavage gene expression and steroid concentration in adult male rats. AB - Adult male rats when treated with 0.4 mg tamoxifen (tam)/kg per day for 90 days show reduced circulating testosterone (T) and LH. The present study was designed to have an in depth understanding of tam induced androgen reduction in adult male rats. Adult male rats were orally administered 0.4 mg tam/kg per day for 30, 60 or 90 days and the temporal effects on intratesticular concentrations of pregnenolone (P(5)), progesterone (P(4)), T, 5 alpha-dihydrotestosterone (5 alpha DHT) and estradiol (E(2)) were estimated. Control group rats were fed saline. Serum hormonal profile of LH, FSH, T and E(2) was also followed on these days. Testicular levels of cytochrome P450 scc mRNA transcripts on 30, 60 and 90 days of treatment with the same dose were quantitated by biplex RT-PCR using beta Actin as internal control followed by analysis using GelPro Analysis software.A significant reduction in intratesticular P(5), P(4), T, 5 alpha-DHT and E(2) was observed from day 30 of treatment. The P450 scc gene expression in the testis was reduced during treatment period from day 60 of treatment. This study demonstrates for the first time that tam reduces testicular pregnenolone biosynthesis through an effect on cholesterol transport and downregulation of P450 scc gene expression. In confirmation of the observed estrogenic effects of tam in this study, it is suggested that E(2) may have a role in cholesterol transport and testicular pregnenolone biosynthesis at the level of cytochrome P450 scc as shown by us. PMID- 12589943 TI - Genomics of steroid hormones: in silico analysis of nucleotide sequence variants (polymorphisms) of the enzymes involved in the biosynthesis and metabolism of steroid hormones. AB - Alterations of steroid hormone biosynthesis and metabolism are suspected to be involved in the pathogenesis of several diseases. Several polymorphisms of the enzymes involved in these processes have already been described and some could be associated with certain diseases. We attempted to examine the sequence variants of these genes in order to find novel variants by an in silico analysis. We analyzed the known human nucleotide sequences of the enzymes p450 side-chain cleavage enzyme, steroid 17-alpha-hydroxylase/17,20-lyase, 3-beta-hydroxysteroid dehydrogenase types 1 and 2, 21-hydroxylase, 11-beta-hydroxylase, aldosterone synthase, aromatase, 11-beta-hydroxysteroid dehydrogenase types 1 and 2, steroid 5-alpha-reductase types 1 and 2, steroid 5-beta-reductase, dehydroepiandrosterone sulfotransferase, 17-beta-hydroxysteroid dehydrogenase types 1-3. The analysis was performed using the National Center for Biotechnology Information Database by the search tool blastn. We found numerous sequence variants in both coding and non-coding sequences. The majority of these sequence variants have already been described, nevertheless, some appear as novel variants. Some of these may also have functional significance. We hypothesize over the possible significance of these findings and briefly review the available literature. PMID- 12589944 TI - Signal transducer and activator of transcription-3 and phosphatidylinositol-3 kinase as coordinate regulators of melanoma cell response to glucocorticoid hormones. AB - STAT proteins act as signal transducers and activators of transcription in cells treated with cytokines or growth factors. Here we analyzed the possible cooperation between STAT3 and phosphatidylinositol-3 kinase (PI-3 kinase) and its involvement in antiproliferative signals induced by glucocorticoid hormones. Treatment of melanoma cells with dexamethasone (DEX) resulted in coexpression of STAT3 activation and increase in the PI-3 kinase protein level. Using plasmids containing JAK2 and STAT3 constructs, we demonstrated that activation of JAK/STAT signaling led to up regulation of PI-3 kinase and enhancement of DEX's ability to increase PI-3 kinase levels in target cells. Prolonged DEX treatment of melanoma cells resulted in constitutive increases in both STAT3 and PI-3 kinase protein levels that were correlated with increased melanoma resistance to antiproliferative hormone action. Similarly, forced expression of both STAT3 and PI-3 kinase in melanoma cells led to enhanced resistance to hormone treatment. Forced expression of PI-3 kinase led to increase in STAT3 activity in a JAK dependent manner, indicating the existence of a feedback regulatory cascade between the JAK/STAT3 and PI-3 kinase pathways. We suggest that protection of melanoma cells from antiproliferative effects of glucocorticoid hormones may be mediated, at least in part, by the constitutive activation of the STAT3/PI-3 kinase signaling pathway. PMID- 12589945 TI - Rat cytochrome P450c17 gene transcription is initiated at different start sites in extraglandular and glandular tissues. AB - In the male rat, the mRNA of the steroidogenic cytochrome P450c17 is expressed extraglandularly in the stomach, duodenum, kidney and liver, throughout the animal's lifespan, as demonstrated by reverse transcriptase and polymerase chain reaction (RT-PCR) analysis with specific primers. Northern analysis indicated that all tissues, except the kidney, contain high levels of such mRNA, but the relative mobility of liver mRNA is slightly less than that of the testis and other tissues. Thus, we analysed their 5'- and 3'-untranslated terminal regions (UTRs) by means of 5'- and 3'-rapid amplification of cDNA ends (RACE) techniques. All tissues utilised the same polyadenylation site as the testis. In the 5'-UTR of liver mRNA, however, we found a distal transcription start site (TSS) located 252b upstream of that used in testicular P450c17 mRNA, which is placed 41b upstream of the first ATG. The 5'-UTR sequence of liver P450c17 cDNA exactly matched the contiguous upstream untranslated region of the gene, suggesting that alternative splicing was not involved in the synthesis of liver P450c17 transcript. The other tissues used the same TSS present in the testis. Nevertheless, a second TSS located 125b upstream of the first ATG was found in the stomach and duodenum. These results show that the transcriptional regulation of the CYP17 gene in the rat is complex and differs between tissues in the use of TSSs. PMID- 12589946 TI - Ab initio electronic structure of the progestogen norethisterone and its 5 alpha derivatives. AB - The steroid 17 alpha-ethynyl-19-nor-4-androsten-17 beta-ol, 3-one (Norethisterone; NET) and its 5 alpha-dihydro (5 alpha-NET), 3 alpha- and 3 beta tetrahydro derivatives (3 alpha,5 alpha- and 3 beta,5 alpha-NET), were comparatively studied by the ab initio quantum mechanics theory. Additionally, 5 alpha-androstan-3 beta,17 beta-diol (ADIOL) was also studied. The Hartree-Fock method and the 6-31G(*) basis set were used to obtain the lowest energy conformation, geometries, electronic structure and physicochemical properties of the steroids. The results showed bond distances and valence angles similar among all steroids, but some differences in dihedral angles in the A-B-ring system were observed. The electronic structure analysis showed that NET has both frontier orbitals that is, the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) located at the C4-C5 pi-bond. In A-ring reduced derivatives, the HOMO was found at the 17 beta-OH and ethynyl groups. In the case of 5 alpha-NET, the LUMO was confined to the A-ring and its C3 carbonyl group while the two NET tetrahydro-reduced derivatives showed the LUMO at the 17 beta-OH and ethynyl groups. The energy changes of the rotational barrier of the 17 beta-OH group suggest that its movement is somewhat restricted by the 17 alpha ethynyl group. Interestingly both groups at C17 form a single electrostatic potential with high electronic density. On the other side, the 19-nor condition increases the A-ring mobility. However, the 3 beta-OH group of 3 beta,5 alpha-NET may rotate without significant energy differences as compared to the same group in ADIOL. The electronic structure of NET and its A-ring reduced derivatives explains in some extent their interaction with androgen and progesterone receptors as well as their selectivity for the estrogen alpha-receptor. PMID- 12589947 TI - 5 alpha-reductase-catalyzed conversion of testosterone to dihydrotestosterone is increased in prostatic adenocarcinoma cells: suppression by 15-lipoxygenase metabolites of gamma-linolenic and eicosapentaenoic acids. AB - Although the androgens, testosterone (T) and its highly active metabolite dihydrotestosterone (DHT) play a role in the development and progression of prostate cancer, the mechanism(s) are unclear. Furthermore, 5 alpha-reductase which catalyze the conversion of T to DHT, has been a target of manipulation in the treatment of prostatic cancer, hence synthetic 5 alpha-reductase activity inhibitors have shown therapeutic promise. To demonstrate that nutrients derived from dietary sources can exert similar therapeutic promise, this study was designed using benign hyperplastic cells (BHC) and malignant tumorigenic cells (MTC) derived from Lobund-Wistar (L-W) rat model of prostatic adenocarcinoma to test the effects of gamma-linolenic acid (GLA), eicosapentaenoic acid (EPA) and their 15-lipoxygenase metabolites on cellular 5 alpha-reductase activity. Our data revealed: (i) that incubation of MTC with [3H]-T resulted in marked conversion to [3H]-DHT when compared to similar incubation with BHC; (ii) that DHT-enhanced activity of 5 alpha-reductase was inhibited 80% by 15S hydroxyeicosatrienoic acid, the 15-lipoxygenase metabolite of GLA, when compared to 55% by 15S-hydroxyeicosapentaenoic acid, the 15-lipoxygenase metabolite of EPA; and (iii) that their precursor fatty acids, respectively, exerted moderate inhibition. Taken together, the study underscores the biological importance of 15 lipoxygenase metabolites of polyunsaturated fatty acids (PUFAs) in androgen metabolism. PMID- 12589948 TI - Investigations into the biosynthetic pathways for classical and ring B unsaturated oestrogens in equine placental preparations and allantochorionic tissues. AB - In on-going studies of 'classical' and ring B-unsaturated oestrogens in equine pregnancy, the products of metabolism of [2,2,4,6,6-2H(5)]-testosterone and [16,16,17-2H(3)]-5,7-androstadiene-3 beta,17 beta-diol with equine placental subcellular preparations and allantochorionic villi have been identified. Using mixtures of unlabelled and [2H]-labelled steroid substrates has allowed the unequivocal identification of metabolites by twin-ion monitoring in gas chromatography-mass spectrometry (GC-MS). Two types of incubation were used: (i) static in vitro and (ii) dynamic in vitro. The latter involved the use of the Oxycell cartridge (Integra Bioscience Systems, St Albans, UK) whereby the tissue preparation was continuously supplied with supporting medium plus appropriate cofactors in the presence of uniform oxygenation. [2H(5)]-Testosterone was converted into [2H(4)]-oestradiol-17 beta, [2H(4)]-oestrone and [2H(3)]-6-dehydro oestradiol-17 alpha in both placental and chorionic villi preparations, but to a greater extent in the latter, confirming the importance of the chorionic villi in oestrogen production in the horse. On the basis of GC-MS characteristics (M(+) m/z 477/482 (as O-methyl oxime-trimethyl silyl ether), evidence for 19 hydroxylation of testosterone was found in static incubations, while the presence of a 6-hydroxy-oestradiol-17 alpha was recorded in dynamic incubations (twin peaks in the mass spectrum at m/z 504/507, the molecular ion M(+)). It was not possible to determine the configuration at C-6. The formation of small, but significant, quantities of [2H(4)]-17 beta-dihydroequilin was also shown, and a biosynthetic pathway is proposed. In static incubations of placental microsomal fractions, the 17 beta-dihydro forms of both equilin and equilenin were shown to be major metabolites of [2H(3)]-5,7-androstadiene-3,17-diol. Using static incubations of chorionic villi, the deuterated substrate was converted into the 17 beta-dihydro forms of both equilin and equilenin, together with an unidentified metabolite (base peak, m/z 504/506). The isomeric 17-dihydroequilins were also obtained using the dynamic in vitro incubation of equine chorionic villi, together with the 17 beta-isomer of dihydroequilenin. Confirmation of the identity of 17 beta-dihydroequilin and 17 beta-dihydroequilenin was obtained by co-injection of the authentic unlabelled steroids with the phenolic fraction obtained from various incubations. Increases in the peak areas for the non deuterated steroids (ions at m/z 414 (17 beta-dihydroequilin) and 412 (17 beta dihydroequilenin) (both as bis-trimethyl silyl ether derivatives) were observed. Biosynthetic pathways for formation of the ring B-unsaturated oestrogens from 5,7 androstadiene-3 beta,17 beta-diol are proposed. PMID- 12589949 TI - Intestinal and hepatic microsomal metabolism of testosterone and progesterone by a 3 alpha-hydroxysteroid dehydrogenase to the 3 alpha-hydroxy derivatives in the channel catfish, Ictalurus punctatus. AB - Intestinal or hepatic microsomes from channel catfish converted [4-14C] testosterone to three major metabolites: 6 beta-hydroxytestosterone, androstenedione and a third metabolite. Formation of the unknown metabolite required NADPH as cofactor. When incubated with 200 microM testosterone, the rate of formation of the unknown metabolite was 265+/-158 pmol/(min mg) protein (mean+/-S.D.) in microsomes from the proximal intestine, 515+/-93 pmol/(min mg) protein in distal intestine and 226+/-42 pmol/(min mg) protein in hepatic microsomes. Comparison of the chromatographic and spectral properties of the unknown metabolite with those of authentic testosterone derivatives showed that this metabolite corresponded to 4-androstene-3 alpha,17 beta-diol. No 3 alpha reduced metabolite was formed in incubations of testosterone with catfish intestinal cytosol. Testosterone was reduced to 5 alpha-dihydrotestosterone primarily in the cytosolic fraction and not in microsomes. Incubation of progesterone with intestinal microsomes resulted in the formation of a metabolite with properties similar to that of the 3 alpha-reduced testosterone, and this metabolite was identified by co-chromatography with authentic standard as 3 alpha reduced progesterone. Thus, 3 alpha-hydroxysteroid dehydrogenase is an important pathway in intestinal microsomes of the channel catfish. PMID- 12589950 TI - Design, synthesis and biochemical evaluation of AC ring mimics as novel inhibitors of the enzyme estrone sulfatase (ES). AB - We report the results of our study into a series of 4'-O-sulfamoyl-4-biphenyl based compounds as novel inhibitors of the enzyme estrone sulfatase (ES). From the results of the molecular modeling design process, it was suggested that these compounds would be able to mimic both the A and C rings of the steroid backbone, and thus possess inhibitory activity against ES. The results of the biochemical evaluation study show that these compounds are indeed good inhibitors, possessing greater inhibitory activity than COUMATE, but weaker inhibitory activity than EMATE or the tricyclic derivative of COUMATE, namely 667-COUMATE. Furthermore, the compounds are observed to be irreversible inhibitors. PMID- 12589951 TI - Intranasal immunization with a lipooligosaccharide-based conjugate vaccine from nontypeable Haemophilus influenzae enhances bacterial clearance in mouse nasopharynx. AB - Nontypeable Haemophilus influenzae (NTHi) is a major cause of otitis media in children. We investigated whether intranasal immunization with a detoxified lipooligosaccharide-tetanus toxoid (dLOS-TT) conjugate vaccine would generate protective immunity against NTHi in a mouse model of nasopharyngeal clearance. The results demonstrated that intranasal immunization with dLOS-TT plus adjuvant cholera toxin (CT) significantly induced LOS-specific IgA antibodies in mouse external secretions, especially in nasal wash (90-fold), bronchoalveolar lavage fluid (25-fold), saliva (13-fold) and fecal extract (three-fold). LOS-specific IgA antibody-forming cells were also found in mucosal and lymphoid tissues with their highest numbers in the nasal passage (528 per 10(6) cells). In addition, the intranasal immunization elicited a significant rise in LOS-specific IgG (32 fold) and IgA (13-fold) in serum. For the immunized mice which had been challenged through the nose with 10(7) live NTHi strain 9274 cells, the vaccine group showed a significant reduction (74-77%) of NTHi, compared to that of control groups with CT alone or dLOS plus CT (P<0.05). Negative correlations were found between bacterial counts and the levels of nasal wash IgA or IgG, saliva IgA and serum IgG. The clearance of five heterologous strains was investigated and revealed a significant clearance of strains 3198, 5657 and 7502 but not of strains 1479 and 2019. These data suggest that intranasal immunization with dLOS TT vaccine elicits both mucosal and systemic immunity against NTHi and enhances bacterial clearance from nasopharynx in mice. Such a vaccine and vaccination regime may be applicable to humans with an appropriate formulation. PMID- 12589952 TI - Interaction of sertraline with Candida species selectively attenuates fungal virulence in vitro. AB - This study investigated whether the interaction between isolates of Candida albicans (n=7), Candida parapsilosis (n=3), Candida krusei (n=2), Candida dubliniensis (n=1) and sertraline, a typical selective serotonin reuptake inhibitor, alters candidal virulence. Sertraline treatment of Candida spp. significantly (P<0.05) affected hyphal elongation, phospholipase activity, production of secreted aspartyl proteinases and fungal viability. In addition, monocyte-derived macrophages (MDMs) treated with sertraline reduced inhibition of blastoconidia germination in comparison to MDMs alone. In conclusion, our findings suggest that the interaction between sertraline and Candida spp. may also diminish the virulence properties of this fungal pathogen in vivo. PMID- 12589953 TI - Eradication of Propionibacterium acnes by its endogenic porphyrins after illumination with high intensity blue light. AB - Propionibacterium acnes is a Gram-positive, microaerophilic bacterium that causes skin wounds. It is known to naturally produce high amounts of intracellular porphyrins. The results of the present study confirm that the investigated strain of P. acnes is capable of producing endogenic porphyrins with no need for any trigger molecules. Extracts from growing cultures have demonstrated emission peaks around 612 nm when excited at 405 nm, which are characteristic for porphyrins. Endogenic porphyrins were determined and quantified after their extraction from the bacterial cells by fluorescence intensity and by elution retention time on high-performance liquid chromatography (HPLC). The porphyrins produced by P. acnes are mostly coproporphyrin, as shown by the HPLC elution patterns. Addition of delta-aminolevulinic acid (ALA) enhanced intracellular porphyrin synthesis and higher amounts of coproporphyrin have been found. Eradication of P. acnes by its endogenic porphyrins was examined after illumination with intense blue light at 407-420 nm. The viability of 24 h cultures grown anaerobically in liquid medium was reduced by less than two orders of magnitude when illuminated once with a light dose of 75 J cm(-2). Better photodynamic effects were obtained when cultures were illuminated twice or three times consecutively with a light dose of 75 J cm(-2) and an interval of 24 h between illuminations. The viability of the culture under these conditions decreased by four orders of magnitude after two illuminations and by five orders of magnitude after three illuminations. When ALA-triggered cultures were illuminated with intense blue light at a light dose of 75 J cm(-2) the viability of the treated cultures decreased by seven orders of magnitude. This decrease in viability can occur even after a single exposure of illumination for the indicated light intensity. X-ray microanalysis and transmission electron microscopy revealed structural damages to membranes in the illuminated P. acnes. Illumination of the endogenous coproporphyrin with blue light (407-420 nm) apparently plays a major role in P. acnes photoinactivation. A treatment protocol with a series of several illuminations or illumination after application of ALA may be suitable for curing acne. Treatment by both pathways may overcome the resistance of P. acnes to antibiotic treatment. PMID- 12589954 TI - Antibody-inducing properties of a prototype bivalent herpes simplex virus/enterotoxigenic Escherichia coli DNA vaccine. AB - The antibody-inducing properties of a bacterial/viral bivalent DNA vaccine (pRECFA), expressing a peptide composed of N- and C-terminal amino acid sequences of the herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) fused with an inner segment encoding the major structural subunit of enterotoxigenic Escherichia coli (ETEC) CFA/I fimbriae (CFA/I), was evaluated in BALB/c mice following intramuscular immunization. The bivalent pRECFA vaccine elicited serum antibody responses, belonging mainly to the IgG2a subclass, against both CFA/I and HSV gD proteins. pRECFA-elicited antibody responses cross-reacted with homologous and heterologous ETEC fimbrial antigens as well as with type 1 and type 2 HSV gD proteins, which could bind and inactivate intact HSV-2 particles. On the other hand, CFA/I-specific antibodies could bind but did not neutralize the adhesive functions of the bacterial CFA/I fimbriae. In spite of the functional restriction of the antibodies targeting the bacterial antigen, the present evidence suggests that fusion of heterologous peptides to the HSV gD protein represents an alternative for the design of bivalent DNA vaccines able to elicit serum antibody responses. PMID- 12589955 TI - Anti-TNF antibody treatment reduces mortality in experimental dengue virus infection. AB - Here we describe a lethal mouse model infected with dengue virus type 2 with several similarities to human DEN-2 infection. Clinically animals demonstrated anemia, thrombocytopenia, pre-terminal paralysis and shock. The most impressive changes were seen with tumor necrosis factor (TNF)-alpha, which abruptly and steeply increased 24 h before the exitus (mean at day 6). Serum levels of IL 1beta, IL-6, IL-10, IL-1 receptor antagonist and soluble TNF receptor I continuously increased during the time of infection. A 100% mortality rate was noted in that group of animals. Treating animals with anti-TNF-alpha serum reduced mortality rate down to 40% (P<0.05). Our model supports the view that activation of innate immune response is at least partially responsible for mortality in DEN-2 infection, and in line with this concept, anti-TNF treatment significantly reduces mortality rates. PMID- 12589956 TI - Isolation and characterization of hemolysin activated by reductant from Prevotella intermedia. AB - The hemolysin from Prevotella intermedia was partially purified from culture supernatant and then characterized. The hemolysin produced a clear beta-hemolytic zone on a blood agar plate. Hemolytic activity was 2.5-fold greater in culture supernatant compared to that cell-associated. The isolation and purification procedure involved ammonium sulfate and polyethylene glycol precipitations and ion-exchange chromatographies on DEAE-Sephacel and CM-Sepharose. The activity of this hemolysin was stimulated by reductants such as cysteine, dithiothreitol, glutathione etc., and was lost upon oxidation. Trypsin or heat treatment resulted in complete inhibition of hemolytic activity. Ca(2+), Mg(2+) and EDTA did not affect the activity. The optimal pH of this hemolysin was 7.5. PMID- 12589957 TI - Pro-inflammatory cytokine and chemokine release by human brain microvascular endothelial cells stimulated by Streptococcus suis serotype 2. AB - Streptococcus suis serotype 2 is a world-wide agent of diseases among pigs including meningitis, septicemia and arthritis. This microorganism is also recognized as an important zoonotic agent. The pathogenesis of the meningitis caused by S. suis is poorly understood. We have previously shown that S. suis is able to adhere to human brain microvascular endothelial cells (BMEC), but not to human umbilical vein endothelial cells (HUVEC). The objective of this work was to study the ability of S. suis serotype 2 to induce the release of the pro inflammatory cytokines tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL 1); IL-6 and the chemokines IL-8 and monocyte chemotactic protein-1 (MCP-1) by human BMEC and HUVEC, using a sandwich enzyme-linked immunosorbent assay. S. suis was able to stimulate the production of IL-6, IL-8 and MCP-1 by BMEC but not HUVEC, in a time- and concentration-dependent manner. Bacterial cell wall components were largely responsible for such stimulation. The human and pig origin of strains does not seem to affect the intensity of the response; indeed, a very heterogeneous pattern of cytokine and chemokine production was observed for the different strains tested in this study. In situ production of cytokines and chemokines by BMEC may be the result of specific adhesion of S. suis to this cell type, with several consequences such as increased recruitment of leukocytes and an increase in the blood-brain barrier permeability. PMID- 12589958 TI - Evaluation in rabbits of different anti-SHIV vaccine strategies based on DNA/fowlpox priming and virus-like particle boosting. AB - Two different prime-boost immunization protocols were tested in rabbits and their immune response was evaluated and compared with the final aim of defining a vaccine strategy that might be able to protect non-human primates from infection with the pathogenic simian/human immunodeficiency virus, SHIV(89.6P). The two regimens were based on three priming immunizations with either an expression plasmid plus a fowlpox (FP) recombinant vector or with two FP recombinant vectors, each one expressing either the SIV(mac239) gag/pol or the HIV 1env(89.6P) genes. In both protocols, priming immunizations were followed by two boosts with SHIV-mimicking virus-like particles (VLP). A complete SHIV-specific response was observed in all animals. Interestingly, the DNA vaccine was three to 10 times more efficient than the FP recombinant in inducing an anti-gag humoral response. Real-time PCR confirmed the memory effect on T-cell subsets secreting interleukin-4 and interferon-gamma, as a consequence of stimulation of both arms of the immune system. Although both protocols were almost equally effective in eliciting homologous neutralizing antibodies and highlighted the efficacy of VLP administration for boosting, protocol A seemed to be more effective in promoting a balanced T-cell memory immune response and appears more promising for vaccine purposes. PMID- 12589959 TI - Beta-glucan, extracted from oat, enhances disease resistance against bacterial and parasitic infections. AB - The effect of beta-glucan, extracted from oats, on the enhancement of resistance to infections caused by Staphylococcus aureus and Eimeria vermiformis was studied in mice. In vitro study using macrophages isolated from the peritoneal cavity showed that beta-glucan treatment significantly enhanced phagocytic activity. In vivo study further demonstrated that beta-glucan treatment induced a significant (P<0.05) protection against the challenge with 5 x 10(8) of S. aureus in mice. Fecal oocyst shedding in the C57BL/6 mice infected with E. vermiformis was diminished by beta-glucan treatment by 39.6% in intraperitoneal and 28.5% in intragastric group compared to non-treated control. Patency period was shorter and antigen (sporozoites and merozoites) specific antibodies were significantly (P<0.05-0.01) higher in beta-glucan-treated group compared to non-treated control group. There were an increasing number of splenic IFN-gamma-secreting cells in glucan-treated group via intraperitoneal route, which might be responsible for the enhancement of the disease resistance. Glucan treatment was able to effectively change the lymphocytes population (Thy 1.2(+), CD4(+) and CD8(+) cells) in the mesenteric lymph nodes and Peyer's patches in mice infected with E. vermiformis. In conclusion, the oral or parenteral oat beta-glucan treatment enhanced the resistance to S. aureus or E. vermiformis infection in the mice. PMID- 12589960 TI - Response of primed human PBMC to synthetic peptides derived from hepatitis B virus envelope proteins: a search for promiscuous epitopes. AB - This investigation was aimed at identifying effective T helper cell epitopes to the hepatitis B virus in humans. A panel of synthetic peptides that represent the hepatitis B virus whole envelope proteins was examined for their capability to stimulate peripheral blood mononuclear cells from human subjects infected with hepatitis B virus naturally. In addition, a large number of subjects were examined and their human leukocyte antigen (HLA) class II allele types were identified to determine whether the helper T cell epitope is specific for a particular HLA allele or 'promiscuous'. The peptides of the amino acid residues 52-67, 110-125, 190-205, and 228-243 appeared to be immunogenic, and particularly, the 52-67 residue was the most promiscuous epitope peptide. These results would contribute to the better understanding of the helper T cell responses to the hepatitis B virus and provide a useful way in designing epitope based vaccines and future therapeutic strategies. PMID- 12589961 TI - Role of kallikrein enzymes in the central nervous system. AB - Kallikreins are a subgroup of the serine protease family of enzymes. Until recently, it was thought that the human kallikrein gene family includes only three members. Over the past 3 years, the human kallikrein gene locus on chromosome 19q13.4 has been characterized. This family includes 15 members for which new nomenclature has been established. A number of kallikreins are expressed in the central nervous system (CNS). Experimental evidence has shown that at least two kallikreins, KLK6 and KLK8, have potential functions in the CNS. KLK8 (neuropsin) is highly expressed in brain tissues and may play a role in brain development, plasticity and response to stress. Of particular interest is the possible involvement of kallikreins in the pathogenesis of Alzheimer's disease (AD). KLK6 (zyme/protease M/neurosin) seems to be down regulated in serum and tissues of Alzheimer's disease patients and may be involved in amyloid metabolism. PMID- 12589962 TI - Soluble transferrin receptor for the evaluation of erythropoiesis and iron status. AB - Iron transport in the plasma is carried out by transferrin, which donates iron to cells through its interaction with a specific membrane receptor, the transferrin receptor (TfR). A soluble form of the TfR (sTfR) has been identified in animal and human serum. Soluble TfR is a truncated monomer of tissue receptor, lacking its first 100 amino acids, which circulates in the form of a complex of transferrin and its receptor. The erythroblasts rather than reticulocytes are the main source of serum sTfR. Serum sTfR levels average 5.0+/-1.0 mg/l in normal subjects but the various commercial assays give disparate values because of the lack of an international standard. The most important determinant of sTfR levels appears to be marrow erythropoietic activity which can cause variations up to 8 times below and up to 20 times above average normal values. Soluble TfR levels are decreased in situations characterized by diminished erythropoietic activity, and are increased when erythropoiesis is stimulated by hemolysis or ineffective erythropoiesis. Measurements of sTfR are very helpful to investigate the pathophysiology of anemia, quantitatively evaluating the absolute rate of erythropoiesis and the adequacy of marrow proliferative capacity for any given degree of anemia, and to monitor the erythropoietic response to various forms of therapy, in particular allowing to predict response early when changes in hemoglobin are not yet apparent. Iron status also influences sTfR levels, which are considerably elevated in iron deficiency anemia but remain normal in the anemia of inflammation, and thus may be of considerable help in the differential diagnosis of microcytic anemia. This is particularly useful to identify concomitant iron deficiency in a patient with inflammation because ferritin values are then generally normal. Elevated sTfR levels are also the characteristic feature of functional iron deficiency, a situation defined by tissue iron deficiency despite adequate iron stores. The sTfR/ferritin ratio can thus describe iron availability over a wide range of iron stores. With the exception of chronic lymphocytic leukemia (CLL) and high-grade non-Hodgkin's lymphoma and possibly hepatocellular carcinoma, sTfR levels are not increased in patients with malignancies. We conclude that soluble TfR represents a valuable quantitative assay of marrow erythropoietic activity as well as a marker of tissue iron deficiency. PMID- 12589963 TI - Protein carbonyl groups as biomarkers of oxidative stress. AB - Oxidative stress, an imbalance toward the pro-oxidant side of the pro oxidant/antioxidant homeostasis, occurs in several human diseases. Among these diseases are those in which high levels of protein carbonyl (CO) groups have been observed, including Alzheimer's disease (AD), rheumatoid arthritis, diabetes, sepsis, chronic renal failure, and respiratory distress syndrome. What relationships might be among high level of protein CO groups, oxidative stress, and diseases remain uncertain.The usage of protein CO groups as biomarkers of oxidative stress has some advantages in comparison with the measurement of other oxidation products because of the relative early formation and the relative stability of carbonylated proteins. Most of the assays for detection of protein CO groups involve derivatisation of the carbonyl group with 2,4 dinitrophenylhydrazine (DNPH), which leads to formation of a stable dinitrophenyl (DNP) hydrazone product. This then can be detected by various means, such as spectrophotometric assay, enzyme-linked immunosorbent assay (ELISA), and one dimensional or two-dimensional electrophoresis followed by Western blot immunoassay. At present, the measurement of protein CO groups after their derivatisation with DNPH is the most widely utilized measure of protein oxidation. PMID- 12589964 TI - Evaluation of cardiac troponin I and T levels as markers of myocardial damage in doxorubicin-induced cardiomyopathy rats, and their relationship with echocardiographic and histological findings. AB - BACKGROUND: Cardiac troponins I (cTnI) and T (cTnT) have been shown to be highly sensitive and specific markers of myocardial cell injury. We investigated the diagnostic value of cTnI and cTnT for the diagnosis of myocardial damage in a rat model of doxorubicin (DOX)-induced cardiomyopathy, and we examined the relationship between serial cTnI and cTnT with the development of cardiac disorders monitored by echocardiography and histological examinations in this model. METHODS: Thirty-five Wistar rats were given 1.5 mg/kg DOX, i.v., weekly for up to 8 weeks for a total cumulative dose of 12 mg/kg BW. Ten rats received saline as a control group. cTnI was measured with Access(R) (ng/ml) and a research immunoassay (pg/ml), and compared with cTnT, CK-MB mass and CK. By using transthoracic echocardiography, anterior and posterior wall thickness, LV diameters and LV fractional shortening (FS) were measured in all rats before DOX or saline, and at weeks 6 and 9 after treatment in all surviving rats. Histology was performed in DOX-rats at 6 and 9 weeks after the last DOX dose and in all controls. RESULTS: Eighteen of the DOX rats died prematurely of general toxicity during the 9-week period. End-diastolic (ED) and end-systolic (ES) LV diameters/BW significantly increased, whereas LV FS was decreased after 9 weeks in the DOX group (p<0.001). These parameters remained unchanged in controls. Histological evaluation of hearts from all rats given DOX revealed significant slight degrees of perivascular and interstitial fibrosis. In 7 of the 18 rats, degeneration and myocyte vacuolisation were found. Only five of the controls exhibited evidence of very slight perivascular fibrosis. A significant rise in cTnT was found in DOX rats after cumulative doses of 7.5 and 12 mg/kg in comparison with baseline (p<0.05). cTnT found in rats after 12 mg/kg were significantly greater than that found after 7.5 mg/kg DOX. Maximal cTnI (pg/ml) and cTnT levels were significantly increased in DOX rats compared with controls (p=0.006, 0.007). cTnI (ng/ml), CK-MB mass and CK remained unchanged in DOX rats compared with controls. All markers remained stable in controls. Analysis of data revealed a significant correlation between maximal cTnT and ED and ES LV diameters/BW (r=0.81 and 0.65; p<0.0001). A significant relationship was observed between maximal cTnT and the extent of myocardial morphological changes, and between LV diameters/BW and histological findings. CONCLUSIONS: Among markers of ischemic injury after DOX in rats, cTnT showed the greatest ability to detect myocardial damage assessed by echocardiographic detection and histological changes. Although there was a discrepancy between the amount of cTnI and cTnT after DOX, probably due to heterogeneity in cross-reactivities of mAbs to various cTnI and cTnT forms, it is likely that cTnT in rats after DOX indicates cell damage determined by the magnitude of injury induced and that cTnT should be a useful marker for the prediction of experimentally induced cardiotoxicity and possibly for cardioprotective experiments. PMID- 12589965 TI - Cathepsin B and H activities and cystatin C concentrations in cerebrospinal fluid from patients with leptomeningeal metastasis. AB - BACKGROUND: Cysteine proteases are involved in the extension of cancer into the subarachnoid space. The presence of cathepsins B and H along with their potent inhibitor cystatin C in the cerebrospinal fluid (CSF) was investigated in patients with leptomeningeal metastasis of cancer (LM). MATERIALS AND METHODS: CSF samples were obtained in 16 cases of LM (10 solid tumors and 6 leukemia or lymphoma) and compared with 11 cancer cases without involvement of the central nervous system, 12 multiple sclerosis cases and 34 healthy volunteers. The activity of the enzymes was measured, their molecular forms were analyzed by the Western blotting, and the concentration of cystatin C was measured by ELISA. Immunohistochemistry of the leptomeningeal tissues was also performed in six autopsy cases of LM. RESULTS: High activities of cathepsins B and H along with decreased cystatin C concentration were observed in CSF of LM as compared with three control groups. Western blot analysis revealed higher concentration of the enzyme protein as well as its active forms in samples with higher enzyme activity. Cells metastasizing leptomeningeal tissue were clearly positive in immunohistochemical staining of cathepsins, indicating active production by tumor cells. CONCLUSION: Production of cathepsins B and H by tumor cells and their high activity along with concomitant decrease of their potent inhibitor, cystatin C, in the CSF might contribute in the process of metastasis and spread of the cancer cells in the leptomeningeal tissues. A high enzyme activity/cystatin C concentration ratio in the CSF could be useful when diagnosing LM in combination with other parameters. PMID- 12589966 TI - Standard CYP2D6 genotyping procedures fail for the CYP2D6*5 and duplication alleles when hair roots are used as a source of DNA. AB - BACKGROUND: Hair roots provide a useful alternative to blood as a source of DNA for genotyping. Besides simple and non-invasive collections, the DNA extraction step is also easy to perform and is fast. The aim of our study is to determine if hair roots can be used to genotype all of the common CYP2D6 alleles for routine screening purposes. METHOD: The study complies with the Declaration of Helsinki. After obtaining informed consents, both blood and hair samples were collected from 92 patients for genotyping of the CYP2D6 gene. PCR was used to detect the following mutations: CYP2D6*1, *3, *4, *5, *9, *10, *17 and duplication gene. The results were compared where hair roots and blood were used as templates for DNA respectively. RESULTS: When blood was used as a source of DNA for genotyping, all of the investigated CYP2D6 alleles were successfully amplified. However, with hair roots, the genes with the larger fragment sizes: CYP2D6*5 and the duplication gene could not be amplified and the bands of other alleles investigated were faint when visualized under UV light. CONCLUSIONS: DNA extraction from hair roots and leucocytes yielded similar results but the DNA extracted from hair roots did not allow successful amplification of the longer genes such as the CYP2D6*5 and the duplication gene. PMID- 12589967 TI - Plasma concentrations of LPL and LCAT are in putative association with females and alcohol use which are independent negative risk factors for coronary atherosclerosis among Japanese. AB - BACKGROUND: Coronary heart disease (CHD) prevalence appears low among Japanese. Analysis of their negative risk factors is therefore important for public health strategy. METHODS: We analyzed the impact on coronary atherosclerosis of sex, alcohol intake, plasma lipoproteins and enzymes to regulate cholesterol transport, lipoprotein lipase (LPL) and lecithin:cholesterol acyltransferase (LCAT) for the 110 patients who underwent coronary angiography, consecutively enrolled by excluding those >70 years or under hypolipidemic-drug treatment. Subgroup combinations compared were males vs. females in non-drinkers, and drinkers vs. non-drinkers in males. RESULTS: Coronary stenosis was less in females and in drinkers, accompanied by high-density lipoprotein (HDL) in the respective comparison. LPL associated with sex (females>males) and LCAT with alcohol intake (drinkers>non-drinkers) although neither enzyme demonstrated direct correlation with coronary stenosis. LPL positively associated only with HDL in most of subgroups and LCAT correlated with low-density lipoprotein (LDL) in all subgroups but with HDL only in males. CONCLUSIONS: Among non-drinkers, females are at lower risk for coronary atherosclerosis than males mainly due to higher HDL in potential association with high LPL, and that drinkers are protected among males also by high HDL that is in apparent association with LCAT. PMID- 12589968 TI - Zymographic analysis of latent and activated forms of matrix metalloproteinase-2 and -9 in synovial fluid: correlation to polymorphonuclear leukocyte infiltration and in response to infection. AB - BACKGROUND: Matrix metalloproteinase-2 and-9 (MMP-2, MMP-9), and gelatinase A and B participate in the degradation of the extracellular matrix proteins in a variety of inflammatory connective tissue diseases including arthritis. METHODS: Synovial fluid was collected by aseptic aspiration from patients with rheumatoid arthritis (RA), osteoarthritis (OA), gout, infected joint, septic arthritis, and systemic lupus erythematosus (SLE). Synovial fluid was subjected to cell count with polymorphonuclear leukocyte (PMN) differential, Gram staining and culture as necessary. MMP-2 and -9 were characterized by substrate gel electrophoresis (gelatin zymography) to resolve latent and activated 'partially proteolyzed' forms. RESULTS: Gelatin zymography revealed that MMP-9 (92, 130, 225 kDa) in synovial fluid was associated with extent of white blood cell infiltration specifically PMNs. In contrast, fibroblast MMP-2 (72 kDa) was present in all synovial fluids irrespective of PMN count. No MMP-9 was detected in the osteoarthritic specimen with low PMN count. Higher PMN count was associated with the presence of activated MMPs, especially in specimens that were confirmed culture positive. Activated synovial fluid MMPs persisted despite resolution of infection. DISCUSSION: Latent and activated MMP-2 and MMP-9 in synovial fluids fluctuate in proportion to PMN infiltration and specifically in response to infection. The presence of activated MMPs post-therapy would suggest that use of specific MMP inhibitors be indicated to eliminate activated MMPs that apparently persist post-infection. PMID- 12589969 TI - Serum lipid concentrations correlate more strongly with total body fat than with body mass index in obese humans. AB - BACKGROUND: To investigate the associations of body mass index (BMI) and total body fat (TBF) vs. blood lipid concentrations, we measured six anthropometric parameters, body fat mass, and serum lipid profiles in 1529 apparently healthy adults. METHODS: TBF was assessed using a body fat analyzer. Serum concentrations of triglyceride, total cholesterol, and low- or high-density lipoprotein cholesterol (LDL-C or HDL-C) were determined by standard enzymatic procedures. RESULTS: Serum lipid concentrations were more strongly correlated with TBF than with BMI or waist circumference in both men and women. The mean concentrations of total cholesterol in the subjects with high fatness (TBF>95th percentile) were significantly higher than those for the subjects with low fatness (TBF<5th percentile; p<0.01), but no significant differences were observed in serum lipid levels between overweights (BMI>95th percentile) and underweights (BMI<5th percentile). The incidence of hypercholesterolemia was significantly higher in the subjects with high fatness (TBF>95th percentile) than the corresponding overweight subjects (BMI>95th percentile; p<0.01). CONCLUSION: TBF is more strongly associated with serum lipid concentrations in adults, at least as compared to BMI. PMID- 12589970 TI - The effects of total laboratory automation on the management of a clinical chemistry laboratory. Retrospective analysis of 36 years. AB - BACKGROUND: Thirty-six years of data and history of laboratory practice at our institution has enabled us to follow the effects of analytical automation, then recently pre-analytical and post-analytical automation on productivity, cost reduction and enhanced quality of service. METHODS: In 1998, we began the operation of a pre- and post-analytical automation system (robotics), together with an advanced laboratory information system to process specimens prior to analysis, deliver them to various automated analytical instruments, specimen outlet racks and finally to refrigerated stockyards. By the end of 3 years of continuous operation, we compared the chemistry part of the system with the prior 33 years and quantitated the financial impact of the various stages of automation. RESULTS: Between 1965 and 2000, the Consumer Price Index increased by a factor of 5.5 in the United States. During the same 36 years, at our institution's Chemistry Department the productivity (indicated as the number of reported test results/employee/year) increased from 10,600 to 104,558 (9.3-fold). When expressed in constant 1965 dollars, the total cost per test decreased from 0.79 dollars to 0.15 dollars. Turnaround time for availability of results on patient units decreased to the extent that Stat specimens requiring a turnaround time of <1 h do not need to be separately prepared or prioritized on the system. CONCLUSIONS: Our experience shows that the introduction of a robotics system for perianalytical automation has brought a large improvement in productivity together with decreased operational cost. It enabled us to significantly increase our workload together with a reduction of personnel. In addition, stats are handled easily and there are benefits such as safer working conditions and improved sample identification, which are difficult to quantify at this stage. PMID- 12589971 TI - A new method to evaluate spontaneous platelet aggregation in type 2 diabetes by Cellfacts. AB - BACKGROUND: The alterations in the functional activities of platelets in diabetes produce an increase of spontaneous platelet aggregation (SPA) and release of platelet-derived microparticles. Platelet-derived microparticles are shed from platelets during activation by high shear stress, collagen and certain agonists. Although the physiologic role of microparticles has been difficult to assess, the characterization of their biological activity is of interest in view of a possible role in hemostasis and coagulation and their reported involvement in thrombotic disease. METHODS: We propose a new, simple method to evaluate spontaneous platelet aggregation and release of platelet-derived microparticles by the Cellfacts analyser (Microbial System Limited (MSL), Coventry, England) that uses electrical sensing flow impedance determination to detect the size particles and the cells in a conductive fluid. Platelet-rich plasma (PRP) from type 2 diabetes was employed for this study. The importance of platelet activating factor (PAF) on spontaneous platelet aggregation was evaluated and the effect of vitamin E and WEB 2086-BS, an antagonist of platelet-activating factor, was measured. RESULTS AND CONCLUSIONS: Data presented show that Cellfacts could be an easy and fast instrument to check the state of platelets in patients with alterations in the functionality of platelets, and to follow the effect of pharmacological therapy on spontaneous platelet aggregation and the release of platelet-derived microparticles. PMID- 12589972 TI - Serum alpha-tocopherol and ascorbic acid concentrations in Type 1 and Type 2 diabetic patients with and without angiopathy. AB - BACKGROUND: Alpha-tocopherol and ascorbic acid form a part of scavenger system influencing the level of oxidative stress in diabetes mellitus. The aim of this study was to evaluate serum concentrations of alpha-tocopherol and ascorbic acid in Type 1 and Type 2 diabetes mellitus and to compare them with the presence of vascular complications as well as with oxidative stress and endothelial dysfunction. METHODS: A total of 38 Type 1 and 62 Type 2 diabetic patients were subdivided into those with and without angiopathy. Serum alpha-tocopherol and ascorbic acid concentrations were estimated in all patients and in 38 healthy persons. Their results were compared with diabetes control, with oxidative stress measured by plasma malondialdehyde and with endothelial dysfunction estimated by serum N-acetyl-beta-glucosaminidase activity. In addition, the differences in biochemical variables were compared between patients with and without angiopathy. RESULTS: Serum alpha-tocopherol related to the sum of cholesterol and triglyceride concentrations (AT/CHT ratio) was significantly lower in diabetic patients with macroangiopathy than in those without vascular changes (p<0.05). Serum ascorbic acid levels were significantly lower only in Type 2 diabetic patients with macroangiopathy as compared with healthy controls as well as with patients without vascular disease (p<0.01). Positive relationship was observed between serum alpha-tocopherol and cholesterol or triglyceride concentrations in both Type 1 and Type 2 diabetic patients. The presence of oxidative stress together with endothelial dysfunction measured by N-acetyl-beta-glucosaminidase activity was accompanied by lower AT/CHT ratio (p<0.005) in Type 2 diabetic patients. CONCLUSION: Diabetic patients with proven angiopathy or with advanced oxidative stress and endothelial dysfunction have significantly lower AT/CHT ratio and ascorbic acid concentration in serum. Their low concentrations may participate at the increased level of oxidative stress in these individuals. PMID- 12589973 TI - Development of immunoassays for type-5 tartrate-resistant acid phosphatase in human serum. AB - BACKGROUND: Tartrate-resistant acid phosphatase (TRAP) is known as a marker of bone resorption. The purpose of this study was the development of a sensitive and specific immunoassay for TRAP. METHODS: We have developed two types of immunoassays, enzyme-linked immunosorbent assay (ELISA) and immunoselective enzyme immunoassay (ISEA) using monoclonal antibodies to recombinant TRAP, for determination of TRAP in human serum. To evaluate assay performance, recovery and dilution tests were performed. Further, we determined serum TRAP levels of patients with secondary hyperparathyroidism at different pH conditions. RESULTS: The detected ranges of ELISA and ISEA were between 0.08 and 5 microg/l and between 0.063 and 4 U/l. Different concentrations of TRAP added were recovered on average at 98.0% in ELISA and 102.9% in ISEA. In the serial dilution test, serum TRAP levels were on average at 101.6% and 109.6% of the expected values in ELISA and ISEA, respectively. The serum TRAP levels of patients with secondary hyperparathyroidism were significantly higher than those of normal controls in ELISA and ISEA. Similar TRAP levels were obtained in the conditions at pH 5.5 and 6.1 in ISEA. CONCLUSION: The present findings suggest that our assay methods are applicable for clinical tests, and strengthen the idea that serum TRAP is useful as a marker for bone resorption. PMID- 12589974 TI - Altered glycosylation of alpha1-acid glycoprotein in patients with inflammation and diabetes mellitus. AB - BACKGROUND: In certain pathophysiological conditions, such as inflammation rheumatoid arthritis and diabetes mellitus (DM), alterations in asparagine-linked glycan (N-glycan) patterns of the acute-phase protein, alpha(1)-acid glycoprotein (AGP), have been reported. In this study, we investigated N-glycan structures of AGP purified from the sera of patients with acute inflammation (n=5), type 2 diabetes mellitus (n=5), and healthy individuals (n=5). METHODS: N-Glycans were released with peptide N-glycosidase F (PNGase F) from denatured AGP and purified with cellulose cartridge. N-glycans were analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOFMS) in combination with exoglycosidase digestion. RESULTS: We revealed increases in bi antennary complex glycans and in alpha1-3 fucosylated bi-, tri-, and tetra antennary glycans and a decrease in tri-antennary glycans in inflammation patients. These results support increases in bindings to concanavalin A (ConA) and Aleuria aurantia lectins (AALs). In diabetic patients, the pathogenesis specific change in N-glycan patterns of AGP was not significant. CONCLUSIONS: The MALDI-TOFMS method is sensitive and suitable for profiling analysis of N-glycans in clinical samples. PMID- 12589975 TI - Analysis of GSTP1-1 polymorphism using real-time polymerase chain reaction. AB - BACKGROUND: Glutathione transferases (GSTs) play an important role in the protection of cells from the products of oxidative stress as well as from several environmental carcinogens. The GSTP1-1 gene class is significantly overexpressed in many human tumors. Four allelic variants have been described for the GSTP1-1 gene (*A, *B, *C, *D) leading to different amino acid substitutions in position 105 and 114 of the protein sequence. The proteins encoded by the different alleles show different abilities to metabolize carcinogens and anticancer agents, suggesting an association between GSTP1 polymorphism and the risk for a variety of cancers as well as between said polymorphism and varying responses to cancer treatments. METHODS: The GSTP1-1 polymorphism was determined using a real-time polymerase chain reaction (PCR) and fluorescence resonance energy transfer with a Light-Cycler Instrument. ARMS was used in the case of *A/*C or *B/*D heterozygosity. We used this method to determine the GSTP1-1 polymorphism in 250 free-living Italian subjects of both sexes. RESULTS: Among 250 subjects representative of an Italian population, we observed the following allelic frequencies: f(A)=0.710, f(B)=0.236, f(C)=0.054 and f(D)=0. The observed phenotypes are in Hardy-Weinberg equilibrium (chi(2)=0.71, df=4, P=0.95). CONCLUSIONS: We have extended and improved a method of GSTP1-1 complete genotyping. This method provides the ability to genotype 30 samples in 2 h and it represents a fast, reliable and automated methodology to determine GSTP1-1 polymorphism in order to perform large-scale population studies. PMID- 12589977 TI - The evaluation of C-reactive protein, homocysteine and vitamin B6 concentrations in Behcet and rheumatoid arthritis disease. PMID- 12589976 TI - Selective priming of Th1-mediated antigen-specific immune responses following oral administration of mixed prescriptions of traditional Korean medicines. AB - BACKGROUND: In previous studies, we showed that oral administration of traditional Korean medicines, Soamsan (SA) and Bo-yang-hwan-o-tang (BHT), modulated antigen-specific immune responses in mice. METHODS: We attempted to strengthen cell-mediated immune responses in mice using two mixed prescriptions composed mainly of components used in SA and/or BHT. The effect of oral administration of the medicines on the induction of antigen-specific immune responses was investigated using hen egg-white lysozyme (HEL) as a model antigen system. RESULTS: Following oral administration, HEL-specific cellular immune responses were enhanced in HEL low-responder mice, and the concentrations of gamma interferon (IFN-gamma), but not interleukin (IL)-4, increased significantly. In addition, the prescriptions decreased the level of HEL-specific antibodies of the immunoglobulin (Ig)G1 subtype, which is associated with helper T lymphocyte (Th2) cell stimulation. Moreover, the presence of the medicines in vitro significantly increased IFN-gamma production from mouse splenocytes, and the magnitude of the increase was closely associated with glycoprotein concentrations. CONCLUSIONS: The Korean prescriptions enhanced anti-HEL-specific cellular immune responses by selectively priming specific subtypes of the helper T cell population. Consequently, they might be useful therapy for patients who need enhanced Th1, or to suppress Th2 immune responses. PMID- 12589978 TI - Differences in coenzyme Q10 content in deltoid and quadriceps muscles. PMID- 12589979 TI - Dehydroepiandrosterone sulfate concentrations in a nonagenarian population. PMID- 12589980 TI - Spectrophotometric bilirubin measurement combined with blood gas analysis. PMID- 12589981 TI - CK-2 antibody (Conan) recognises cancer associated ectopic form of CK-2. PMID- 12589982 TI - Retraction of published paper. Detection of human apolipoprotein E genotypes by DNA biosensors coupled with PCR. PMID- 12589984 TI - The resuscitation greats. Henning Ruben MD, FFARCS(I), FFARCS. The Ruben valve and the AMBU bag. PMID- 12589985 TI - The physiology of sudden violent death. AB - BACKGROUND AND PURPOSE: The physiology behind sudden violent death is considered in the light of information from a wide variety of clinical, experimental, forensic and veterinary sources. Physiological causes can be classified under the headings of blood loss; asphyxia; electrocution; tissue loss; destruction of brain; disconnection of brain; and poisoning. Death, although sudden, is not necessarily instantaneous; asphyxia and electrocution take some time to kill the subjects. CONCLUSIONS: The chemistry of dying, death and changes post mortem, needs more research in order to design further rational resuscitation procedures. PMID- 12589986 TI - The Public Access Defibrillation (PAD) trial: study design and rationale. AB - The PAD Trial is a prospective, multicenter, randomized clinical study testing whether volunteer, non-medical responders can improve survival from out-of hospital cardiac arrest (OOH-CA) by using automated external defibrillators (AEDs). These lay volunteers, who have no traditional responsibility to respond to a medical emergency as part of their primary job description, will form part of a comprehensive, integrated community approach to the treatment of OOH-CA. The study is being conducted at 24 field centers in the United States and Canada. Approximately 1000 community units (e.g. apartment or office buildings, gated communities, sports facilities, senior centers, shopping malls, etc.) were randomized to treatment by trained laypersons who will provide either cardiopulmonary resuscitation (CPR) alone or CPR plus use of an AED, while awaiting arrival of the community's emergency medical services responders. The primary endpoint is the number of OOH-CA victims who survive to hospital discharge. Secondary endpoints include neurological status, health-related quality of life (HRQL), cost, and cost-effectiveness. Data collection will last approximately 15 months and is expected to be completed in September 2003. PMID- 12589987 TI - Public access defibrillation in Helsinki--costs and potential benefits from a community-based pilot study. AB - In cardiac arrest the interval between the collapse and defibrillation may be shortened by teaching lay people to use defibrillators. We conducted a 3-year prospective, community-based study on public access defibrillation (PAD) in an urban emergency medical services system. All public sites with a cardiac arrest incidence of at least one per year were equipped with automated external defibrillators. Twenty cardiac arrest patients were enrolled, seven in PAD and 13 in control group. Defibrillation was accomplished significantly earlier (P=0.01) in the PAD group. The direct costs were 110,270 Eur and only 13.5-16% of this figure would be related to the cost of defibrillators during their 8 years lifespan. This study showed that a community based model of PAD shortens the time to CPR and defibrillation significantly in an urban environment but various challenges have to be solved before wider implementation of PAD. In future projects the nature of the costs especially should be considered. PMID- 12589988 TI - The assessment of three methods to verify tracheal tube placement in the emergency setting. AB - We studied prospectively the reliability of clinical methods, end-tidal carbon dioxide (ETCO(2)) detection, and the esophageal detector device (EDD) for verifying tracheal intubation in 137 adult patients in the emergency department. Immediately after intubation, the tracheal tube position was tested by the EDD and ETCO(2) monitor, followed by auscultation of the chest. The views obtained at laryngoscopy were classified according to the Cormack grade. Of the 13 esophageal intubations that occurred, one false-positive result occurred in the EDD test and auscultation. In the non-cardiac arrest patients (n=56), auscultation, the ETCO(2), and EDD test correctly identified 89.3, 98.2*, and 94.6%* of tracheal intubations, respectively (*, P<0.05 vs. the cardiac arrest patients). In the cardiac arrest patients (n=81), auscultation, the ETCO(2), and the EDD tests correctly identified 92.6**, 67.9, and 75.3% of tracheal intubations, respectively (**, P<0.05 vs. EDD and ETCO(2)). The frequencies of Cormack grade 1 or 2 were 83.9% in the non-cardiac arrest, and 95.1% in the cardiac arrest patients. In conclusion, the ETCO(2) monitor is the most reliable method for verifying tracheal intubation in non-cardiac arrest patients. During cardiac arrest and cardiopulmonary resuscitation, however, negative results by the ETCO(2) or the EDD are not uncommon, and clinical methods are superior to the use of these devices. PMID- 12589989 TI - The influence of new guidelines on cardiopulmonary resuscitation (CPR) decisions. Five cycles of audit of a clerk proforma which included a resuscitation decision. AB - Current guidelines advise discussion with patients before issuing a 'do not attempt resuscitation' (DNAR) order. We report five audit cycles of cardiopulmonary resuscitation (CPR) documentation after introducing a proforma, the last cycle following the latest guidelines. In first audit data were collected from 75 patient discharges. CPR decisions were documented in 27 (36%). Four subsequent point prevalence audits carried out on all inpatients following proforma introduction showed documentation improved to 102/109 (94%), 135/148 (91%), 131/140 (94%) and 102/119 (86%) in cycles two, three, four and five, respectively. The last three audits also revealed that consultants consistently made more DNAR orders than trainee doctors. However, following the introduction of the latest guidelines the proportion of patients in whom a decision was made, and the percentage of those decisions that were DNAR, fell. PMID- 12589990 TI - Evaluation of a defibrillator-basic cardiopulmonary resuscitation programme for non medical personnel. AB - To improve the outcome for out-of-hospital patients with ventricular fibrillation/pulseless ventricular tachycardia (VF/VT), the use of automated external defibrillators (AEDs) by first responders including non-medical personnel with a duty to respond to an emergency is recommended. A special CPR AED course has been developed. We wanted to test the results (quality and speed of operating an AED and CPR) after completion of such a course and retention after approximately 1-year. At the same time we wanted to see if personnel could use an AED after receiving written information without having attended the course. Study subjects were divided randomly into groups, and tested pre-course (n=54), post-course (n=50), and unannounced 10+/-3 months after the course (retention group, n=61). For statistical analysis two sample tests for binomial proportions and Wilcoxon-Mann-Whitney test was used as appropriate. Fifteen of the 27 pairs (56%) in the pre-course group with no previous exposure to an AED decided to use it. There was no difference between the groups in electrode pad positioning, and all stayed clear of the manikin during the process of AED charging and shock delivery. The post-course group had a higher rate of checking for responsiveness (vs. pre-course), not to check for a pulse (vs. both other groups), the shortest time interval from arrival on scene to start of CPR and shock delivery, and in parallel the shortest hands-off interval (without chest compressions and ventilations) before shock delivery. The quality of chest compressions was improved by the course but decreased to a similar standard as in the pre-course when tested 10+/-3 months later, except for correct depth which was similar to post course. Most ventilation attempts in all groups were scored as incorrect due to the high incidence of excessively rapid inflations. The retention group had a lower frequency of correct inflations than the pre-course group, and the post-course group the highest number of correct ventilations per minute. These findings suggest that use of an AED by untrained laypersons may be feasible and that complex and time-consuming training programmes may not be necessary. The present study also supports the need for annual training and recertification. PMID- 12589991 TI - 'Event tree' analysis of out-of-hospital cardiac arrest data: confirming the importance of bystander CPR. AB - OBJECTIVE: The British National Service Framework (NSF) for heart disease commended the 'Utstein style' for auditing out-of-hospital cardiac arrests. The NSF also set standards for pre-hospital treatment and response times. To increase the flexibility of Utstein, an 'event tree' technique is proposed as an audit tool. Event trees consist of nodes and branches on which numbers, percentages or probability values are entered. METHODS: Using the London Ambulance Service's (LAS) 1997 database on 3,759 out-of-hospital cardiac arrests, 2,772 arrests witnessed by lay bystanders or unwitnessed were analysed focusing on bystander cardiopulmonary resuscitation (BCPR) and response times. RESULTS: The Utstein template showed that witnessed arrests in ventricular fibrillation (VF) or ventricular tachycardia (VT) who had received BCPR achieved a return of spontaneous circulation (ROSC) in the field significantly more often than non BCPR recipients-26 versus 16% (P=0.006). But the likelihood of being admitted to a hospital bed, and discharged alive, was only marginally better for BCPR recipients. To examine the influence of BCPR on the presenting rhythm an event tree showed that in 48% of witnessed BCPR cases the presenting rhythm was VF/VT, whereas, for witnessed non-BCPR cases, 27% were in VF/VT (P<0.0001). With unwitnessed arrests, 31% of BCPR cases were in VF/VT compared with 18% for non BCPR cases (P<0.0001). Call to scene time was less than 8 min for 66% of all VF/VT arrests. CONCLUSION: The event trees, when combined with the Utstein template, demonstrated the importance of examining comprehensively datasets for both witnessed and unwitnessed cardiac arrests when monitoring performance standards. The analyses also emphasised the relevance of community programmes in Greater London for teaching basic life saving skills. PMID- 12589992 TI - Successful intraosseous infusion in the critically ill patient does not require a medullary cavity. AB - OBJECTIVES: To demonstrate that successful intraosseous infusion in critically ill patients does not require bone that contains a medullary cavity. DESIGN: Infusion of methyl green dye via standard intraosseous needles into bones without medullary cavity-in this case calcaneus and radial styloid-in cadaveric specimens. SETTING: University department of anatomy. PARTICIPANTS: Two adult cadaveric specimens. MAIN OUTCOME MEASURES: Observation of methyl green dye in peripheral veins of the limb in which the intraosseous infusion was performed. RESULTS: Methyl green dye was observed in peripheral veins of the chosen limb in five out of eight intraosseous infusions into bones without medullary cavity calcaneus and radial styloid. CONCLUSIONS: Successful intraosseous infusion does not always require injection into a bone with a medullary cavity. Practitioners attempting intraosseous access on critically ill patients in the emergency department or prehospital setting need not restrict themselves to such bones. Calcaneus and radial styloid are both an acceptable alternative to traditional recommended sites. PMID- 12589993 TI - Lunar phases are not related to the occurrence of acute myocardial infarction and sudden cardiac death. AB - BACKGROUND: Mass media deliver pertinacious rumours that lunar phases influence the progress and long-term results in several medical procedures. Peer reviewed studies support this, e.g. in myocardial infarction, others do not. METHODS: We looked retrospectively at the dates of cardiac arrests (CA; n=368) of cardiac origin and of acute myocardial infarctions (AMI) with consecutive thrombolytic therapy or acute PTCA (n=872) and at the lunar phases at the corresponding dates. Medical data had been collected prospectively on the patient's admission. The lunar phases were defined as full moon+/-1 day, new moon+/-1 day and the days in between as waning and waxing moon. The incidence of these cardiac events at each phase was calculated as days with a case divided by the total number of days of the specific moon phase in the observation period (1992-1998). Wilcoxon Rank Test was used for statistical analysis. RESULTS: AMI and CA occurred on equal percentages of days within each lunar phase: AMI on 35% of all days with new moon, on 38% of full moon days, on 39% waning, and on 41% of the waxing moon days; CA on 19, 17, 16 and 16% of all days of the respective lunar phase. This difference was not significant. CONCLUSION: Lunar phases do not appear to correlate with acute coronary events leading to myocardial infarction or sudden cardiac death. PMID- 12589994 TI - Optimizing bag-valve-mask ventilation with a new mouth-to-bag resuscitator. AB - When ventilating an unintubated patient with a self-inflating bag, high peak inspiratory flow rates may result in high peak airway pressure with subsequent stomach inflation; this may occur frequently when rescuers without daily experience in bag-valve-mask ventilation need to perform advanced airway management. The purpose of this study was to assess the effects of a newly developed self-inflating bag (mouth-to-bag resuscitator; Ambu, Glostrup, Denmark) that limits peak inspiratory flow. A bench model simulating a patient with an unintubated airway was used, consisting of a face mask, manikin head, training lung (lung compliance, 100 ml/0.098 kPa (100 ml/cm H(2)O)); airway resistance, 0.39 kPa/l per second (4 cm H(2)O/l/s), oesophagus (LESP, 1.96 kPa (20 cm H(2)O)) and simulated stomach. Twenty nurses were randomised to ventilate the manikin for 1 min (respiratory rate: 12 per minute) with either a standard self-inflating bag or the mouth-to-bag resuscitator, which requires the rescuer to blow up a single use balloon inside the self-inflating bag, which in turns displaces air towards the patient. When supplemental oxygen is added, ventilation with up to 100% oxygen may be obtained, since expired air is only used as the driving gas. The mouth-to-bag resuscitator therefore allows two instead of one hand sealing the mask on the patient's face. The volunteers were blinded to the experimental design of the model until completion of the experimental protocol. The mouth-to bag resuscitator versus standard self-inflating bag resulted in significantly (P<0.05) higher mean+/-S.D. mask tidal volumes (1048+/-161 vs. 785+/-174 ml) and lung tidal volumes (911+/-148 vs. 678+/-157 ml), longer inspiratory times (1.7+/ 0.4 vs. 1.4+/-0.4 s), but significantly lower peak inspiratory flow rates (50+/-9 vs. 62+/-13 l/min) and mask leakage (10+/-4 vs. 15+/-9%); peak inspiratory pressure (17+/-2 vs. 17+/-2 cm H(2)O) and stomach tidal volumes (16+/-30 vs. 18+/ 35 ml) were comparable. In conclusion, employing the mouth-to-bag resuscitator during simulated ventilation of an unintubated patient in respiratory arrest significantly decreased inspiratory flow rate and improved lung tidal volumes, while decreasing mask leakage. PMID- 12589995 TI - Magnesium reduces free radical concentration and preserves left ventricular function after direct current shocks. AB - OBJECTIVE: Our objective was to determine if magnesium reduces free radicals generated by direct current countershock and preserves left ventricular contractile function. BACKGROUND: We have previously shown that magnesium reduces free radicals in a coronary occlusion-reperfusion model, and therefore also might reduce free radical generation by direct current shocks. METHODS: In eight swine weighing 18-27 kg (mean: 22 kg), using electron paramagnetic resonance, we monitored continuously the coronary sinus concentration of ascorbate free radical, a measure of free radical generation (total oxidative flux). Epicardial shocks (30 J) using a truncated exponential biphasic waveform (5/5 ms) were administered. Each animal received two shocks, one without and one with magnesium, 80 mg/min IV, beginning 10 min before the shock and continuing to 15 min after the shock. Percent fractional area shortening of the left ventricular cavity was determined by 2-dimensional echocardiography. RESULTS: Magnesium shocks resulted in a significantly lower increase in the ascorbate free radical concentration (0.6+/-4.6%) than no-magnesium shocks (16+/-3.3%, P<0.05) at 12 min after the shock. Total radical flux was reduced 72% (P<0.05), and left ventricular fractional area shortening was preserved: baseline: 69+/-2.6%, no magnesium shocks: 41+/-2.8% (P<0.05, versus baseline) and magnesium shocks 61+/ 3.7%. CONCLUSIONS: Magnesium pre-treatment reduced oxygen free radicals generated by direct current shocks; post-shock left ventricular contractile function was not impaired. Magnesium may be cardioprotective during epicardial ('surgical') defibrillation. PMID- 12589996 TI - LF 16-0687 Ms, a new bradykinin B2 receptor antagonist, decreases ex vivo brain tissue prostaglandin E2 synthesis after closed head trauma in rats. AB - OBJECTIVE: Bradykinin (B) contributes to secondary brain injury. This injury is mediated in part by prostaglandin (PG). Antagonism of B(2) receptors improves neurological status after brain injury, but the effect of B(2) antagonism on brain tissue PG is unknown. This study examined the effect of LF 16-0687 Ms, a new B(2) receptor antagonist, on brain tissue PGE(2) after closed head trauma (CHT). METHODS: Rats were anesthetized and received sham+saline, sham+LF 16-0687 Ms, CHT+saline, or CHT+LF 16-0687 Ms. Brain tissue samples were obtained at 24 h for determination of PGE(2) (after 2 h of ex vivo incubation) and water content. Neurological severity score (NSS) was assessed at 1 and 24 h. RESULTS: In the group receiving CHT+LF 16-0687 Ms, brain tissue PGE(2) (77.7+/-65.9 pg/mg tissue, mean+/-SD) was less than in the group receiving CHT+saline (368.1+/-186.2 pg/mg tissue) and not different than sham+saline (78.7+/-30.7 pg/mg tissue). LF 16-0687 Ms also improved NSS and decreased brain water content by 51%. CONCLUSION: We conclude that the beneficial effect of LF 16-0687 Ms on outcome after CHT is accompanied by blockade of PGE(2) increase in injured brain tissue. PMID- 12589997 TI - Survival and normal neurological outcome after CPR with periodic Gz acceleration and vasopressin. AB - BACKGROUND: We showed previously that whole body periodic acceleration along the spinal axis (pGz) is a novel method of cardiopulmonary resuscitation (CPR). The ultimate assessment of the value of any CPR technique is the neurological outcome after using such a technique. In this study, we determined the neurological outcome in pigs after prolonged pGz-CPR, with administration of vasopressin immediately prior to defibrillation. Neurological outcome after pGz-CPR was compared to a control group where no intervention occurred for the same time period (C-NoInterv). METHODS AND RESULTS: Ventricular Fibrillation (VFIB) was induced in 12 animals. After a 3 min non-interventional interval, the animals received either pGz-CPR (n=7), or C-NoInterv (n=5) for 15 min. After 18 min of VFIB, a single dose of vasopressin (0.8 U/kg) was administered along with sodium bicarbonate and bretylium, and defibrillation was attempted. All animals in the pGz-CPR group had return of spontaneous circulation (ROSC) and normal neurological assessment at 24 h. Neurologic outcome remained normal at 48 h. In contrast, none of the animals in the C-NoInterv had ROSC. CONCLUSION: Prolonged pGz-CPR, with administration of vasopressin immediately prior to defibrillation results in normal neurological outcomes at 24 h. PMID- 12589999 TI - Re-evaluation of cardiopulmonary resuscitation. PMID- 12589998 TI - Effects of vasopressin on adrenal gland regional perfusion during experimental cardiopulmonary resuscitation. AB - OBJECTIVE: Despite the important role of the adrenal gland during cardiac arrest, little is known about changes in the adrenal medullary or cortical blood flow in this setting. This study was designed to assess regional adrenal gland perfusion in the medulla and cortex during cardiopulmonary resuscitation (CPR), and after administration of adrenaline (epinephrine) versus vasopressin versus saline placebo. METHODS: After 4 min of untreated ventricular fibrillation, and 3 min of basic life support CPR, 19 animals were randomly assigned to receive either vasopressin (0.4 U/kg; n=7), adrenaline (45 microg/kg; n=6) or saline placebo (n=6), respectively. Haemodynamic variables, adrenal, and renal blood flow were measured after 90 s of CPR, and 90 s and 5 min after drug administration. RESULTS: All values are given as mean+/-S.E.M. Blood flow in the adrenal medulla was significantly higher 90 s after adrenaline when compared with saline placebo in the right adrenal medulla (210+/-14 vs. 102+/-5 ml/min per 100 mg), and in the left adrenal medulla (218+/-14 vs. 96+/-3 ml/min per 100 mg). Blood flow in the adrenal medulla was significantly higher 90 s and 5 min after vasopressin when compared with adrenaline in the right (326+/-22 mg vs. 210+/-14 ml/min per 100 mg, and 297+/-17 vs. 103+/-5 ml/min per 100 mg), and in the left medulla (333+/ 25 vs. 218+/-14 ml/min per 100 mg, and 295+/-14 vs. 111+/-7 ml/min per 100 mg). Ninety seconds and five minutes after vasopressin, and 90 s after adrenaline, adrenal cortex blood flow was significantly higher when compared with saline placebo. After 12 min of cardiac arrest, including 8 min of CPR, seven of seven pigs in the vasopressin group, one of six pigs in the adrenaline group, but none of six placebo were successfully defibrillated. CONCLUSION: Both vasopressin and adrenaline produced significantly higher medullary and cortical adrenal gland perfusion during CPR than did a saline placebo; but vasopressin resulted in significantly higher medullary adrenal gland blood flow when compared with adrenaline. PMID- 12590001 TI - Long QT: the chicken or the egg. PMID- 12590002 TI - Newborn manikins. PMID- 12590004 TI - Weight control and the management of obesity after menopause: the role of physical activity. AB - BACKGROUND: Postmenopausal women have an increased tendency for gaining weight. The declines of endogenous estrogen, together with physical inactivity, are probably the major causes of this phenomenon. Postmenopausal overweight and obesity leads to increased rates of hypertension, diabetes mellitus, coronary artery disease, and all cause mortality. Additional consequences may include hormone-dependent cancer, gallstones, nephrolithiasis, and osteoarthritis. Weight loss can reverse many of these complications, reduce the number and dosages of medications used, and improve longevity. This difficult task requires lifestyle modification. OBJECTIVES: To review the current information about the effects of physical activity on postmenopausal weight gain and its consequences and to provide basic strategies to treat obesity during the menopause transition. METHODS: A Medline search together with a manual search of selected articles. CONCLUSION: Several options for weight loss are available, yet lifestyle modification is essential in managing postmenopausal obesity and overweight. While this demands dietary and behavioral changes, a major element of this modification is regular physical activity, which reduces obesity-related morbidity and mortality. The amendment to a healthier lifestyle is achievable at the postmenopausal years. All medical personnel should advocate against overweight and obesity and provide tools for their management. PMID- 12590005 TI - Kava-Kava administration reduces anxiety in perimenopausal women. AB - OBJECTIVE: Disturbances of mood, such as anxiety and depression, increase in the perimenopausal period. Hormone replacement therapy or neuroactive drugs represent useful treatments for these disturbances but may be contraindicated or not accepted. Herein it was investigated the efficacy of Kava-Kava, an extract of Piper Methysticum, on mood of perimenopausal women. DESIGN: A 3-months randomized prospective open study investigating in perimenopausal women modifications induced by calcium supplementation (control; n=34), calcium plus Kava-Kava at the dose of 100 mg/day (n=15) or calcium plus Kava-Kava at the dose 200 mg/day (n=19). Anxiety was evaluated by the State Trait Anxiety Inventory (STAI); depression by the Zung's scale (SDS), and climacteric symptoms by the Greene's scale. Evaluations were performed at baseline and after 1 and 3 months. RESULTS: In the control group during the 3 months, anxiety, depression and climacteric symptoms tended to decline, but not significantly. During Kava-Kava anxiety declined (P<0.001) at 1 (-3.8+/-1.03) and 3 (-5.03+/-1.2) months, depression declined at 3 months (-5.03+/-1.4; P<0.002) and climacteric score declined (P<0.0006) at 1 (-2.87+/-1.5) and 3 (-5.38+/-1.3) months. Only the decline of anxiety induced by Kava-Kava was significantly greater than that spontaneously occurring in controls (P<0.009). CONCLUSIONS: The present data indicate that, in perimenopausal women, administration of Kava-Kava induces an improvement of mood, particularly of anxiety. PMID- 12590006 TI - Response to alendronate in osteoporotic women previously treated with pamidronate. AB - INTRODUCTION: Different bisphosphonates have been shown to increase bone mineral density (BMD) and reduce the risk of fracture in osteoporotic patients. It is unclear how shifting from a treatment with one bisphosphonate to another will influence the evolution of BMD and bone turnover. METHODS: In the present study, we followed BMD (DXA, Hologic QDR1000) of the lumbar spine (BMDL) and of the total hip (BMDH), bone alkaline phosphatase (Ostase, Hibritech), and urinary collagen cross links (pyridinoline, deoxypyridinoline, Biorad) in 39 patients treated with IV pamidronate (60 mg/3 months) since at least 2 years and who were shifted to oral alendronate (10 mg/day, n=18) or left to IV pamidronate (n=21) for 2 more years. RESULTS: BMD increased similarly and significantly in both groups after 2 additional years of treatment as compared to baseline (P<0.05, sign test). BMDL: +3.8% in the alendronate group vs +4.1% in the pamidronate group; BMDH: +4.3% in alendronate group vs +3.6% in pamidronate group, There was no significant change in the biological parameters of bone turnover in any group. CONCLUSION: The increase of BMD with both bisphosphonates in these previously treated patients was as expected after a 2 more years of treatment. Alendronate administration did not induce a larger gain in BMD as compared to cyclic pamidronate. Bone turnover was no longer affected by switching the bisphosphonate treatment. PMID- 12590008 TI - Rapid changes of flow-mediated dilatation after surgical menopause. AB - OBJECTIVES: Estrogen acts directly on endothelial nitric oxide synthase through a non-genomic mechanism, resulting in rapid dilatation of blood vessels. In this study, we examined the change of endothelial function after surgical menopause. METHODS: In 20 subjects who underwent gynecological operations (ovariectomy (OVX) 12, sham (SHAM) operation 8), postoperative changes of flow-mediated dilatation (FMD) of the brachial artery were examined using ultrasonography. Postoperative changes of the response to nitroglycerin (NTG) were also studied in these patients. RESULTS: In the OVX group, significant decreases of FMD were observed 1 week after the operation, although no changes were observed in the response to NTG. In the SHAM group, no remarkable changes of FMD or the response to NTG were observed after the operation. CONCLUSIONS: OVX influences endothelium-dependent vasodilatation within as little as 1 week. Therefore, it may be important to address the rapid changes of circulation after surgical menopause in order to prevent cardiovascular disease. PMID- 12590007 TI - Endothelin-1 and nitric oxide levels are related to cardiovascular risk factors but are not modified by estradiol replacement in healthy postmenopausal women. A cross-sectional and a randomized cross-over study. AB - OBJECTIVE: To evaluate whether in healthy postmenopausal women endothelial substances such as endothelin-1 (ET-1) and nitric oxide are related to cardiovascular risk factors and can be influenced by estradiol replacement. DESIGN: A cross-sectional evaluation and a randomized, double-blind, placebo controlled study with cross-over. METHODS: In 20 healthy postmenopausal women it was investigated the relation of ET-1 and NOx with age, BMI, 24-h blood pressure, lipid and glucose metabolism, and coagulation parameters. In addition, in the same women, the role played by estrogens on circulating ET-1 and stable derivatives of nitric oxide (nitrite/nitrates) was investigated by administering for 2 months transdermal estradiol (50 microg/day) vs. placebo. RESULTS: ET-1 and NOx were inversely related to each other (r=0.458; P=0.016). Multivariate analysis of regression showed that ET-1 levels were related directly to LDL cholesterol (r=0.585; P=0.0005) and protein C (r=0.516; P=0.0008), and inversely to insulin (r=0.488; P=0.0065). The ratio NOx/ET-1 was directly related to HDL cholesterol (r=0.441; P=0.005). The above relations were not influenced by estradiol. Indeed, in comparison to placebo, transdermal estradiol, besides reducing nocturnal systolic (P=0.002) and diastolic (P=0.03) blood pressure, did not modify ET-1 or NOx levels, as well as, any of the parameters considered. CONCLUSIONS: The relation of several cardiovascular risk factors with ET-1 and NOx/ET-1 suggests a primary role for these endothelial products in the determination of the cardiovascular risk of women. The present data do not support a role for transdermal estradiol in modifying ET-1 or NOx levels of healthy postmenopausal women. PMID- 12590009 TI - Is hormone replacement therapy associated with an increased risk of irritable bowel syndrome? AB - OBJECTIVE: Hormonal status could be involved in the occurrence of irritable bowel syndrome (IBS). The authors examined the risk of developing IBS in women using hormone replacement therapy (HRT). METHODS: Women 50-69 years old with at least one prescription for HRT during 1994-1999 were identified from the General Practice Research Database in the UK (n=40, 119). An aged-matched cohort of 50000 women who never used HRT was sampled from the source population where the HRT cohort was ascertained. Women in the two cohorts were followed to assess the risk of development of IBS. Authors performed a nested case-control analysis to assess the role of duration, route and regimen of HRT use and other risk factors for IBS. The IBS diagnosis was validated by means of a questionnaire sent to the general practitioners (n=660). RESULTS: The incidence rate of IBS per 1000 person years was 1.7 in the cohort of never HRT users and 3.8 among HRT users, respectively. Both current and past users of HRT presented an increased risk of IBS compared to non-users, after adjusting for co-morbidity and consultation patterns. This increased risk was observed irrespective of treatment duration, regimen or route of administration of HRT. CONCLUSION: The result suggests that HRT use is associated with an increased risk of IBS similar to the one observed among younger premenopausal women with endogenous oestrogenic activity. PMID- 12590010 TI - The influence of serum leptin concentration on bone mass assessed by quantitative ultrasonometry in pre and postmenopausal women. AB - OBJECTIVE: the aim of this study was to evaluate the influence of serum leptin concentration on bone mass assessed by quantitative ultrasound (QUS) in a large sample of healthy pre and postmenopausal women. DESIGN: 555 healthy pre and postmenopausal (n=261 and n=294) women (mean age, 49.5+/-17.2 years) not on hormone replacement therapy were recruited on the occasion of a routine gynecological visit. Before entry to the study, all women had answered a detailed questionnaire on important risk factors and gave written informed consent. Speed of sound (SOS), broadband ultrasound attenuation (BUA) and stiffness index (SI) of the os Calcis were measured using the Achilles ultrasonometer (GE/lunar). We systematically investigated the relation of menopause, BMI and leptin on bone mass by allocating women into the following groups: (a) premenopausal women BMI<25 kg/m(2) (N=178); (b) premenopausal women BMI>25 kg/m(2) (N=83); (c) postmenopausal women BMI<25 kg/m(2) (N=125); and (d) postmenopausal women BMI>25 kg/m(2) (N=169). Additionally we investigated the relation of serum leptin concentrations, age and BMI on ultrasonometry variables by performing a multiple linear regression analyses. RESULTS: in the initial analyses premenopausal women showed a significantly (P<0.001) lower mean age, weight, BMI, follicle stimulating hormone (FSH) and leptin concentration, a higher mean height, serum estradiol and ultrasonometry variables in comparison to postmenopausal women. Irrespective of the menopausal status, women with a BMI>25 kg/m(2) had significantly higher leptin concentrations (P<0.001) and BUA (P<0.05) whereas SOS and SI was not significant different, compared to women with a BMI<25 kg/m(2). The multiple linear regression analyses showed that only BMI but not Leptin was related to higher ultrasonometry variables, whereas increasing age was associated with a decrease in ultrasonometry variables. Furthermore, the multiple linear regression analyses confirmed that age and BMI were the only statistically significant independent predictor for ultrasonometry variables. There was no significant influence of leptin on ultrasonometry variables even after controlling for BMI or age, or BMI and age. CONCLUSIONS: serum leptin concentrations are significantly higher in pre and postmenopausal obese women, compared with normal weight controls. Ultrasonometry variables are influenced by age and BMI but not by serum leptin concentrations. PMID- 12590011 TI - Factors associated with menopausal symptom reporting in Chinese midlife women. AB - OBJECTIVES: this paper examines whether menopausal status, sociodemographic factors, self-perceived health, and social stress are related to symptom reporting in a community-based sample of Chinese women in midlife. METHODS: a random telephone survey of perimenopausal women aged 44-55 years was conducted in 1996. Eligible subjects were identified through telephone dialing of a random sample of the numbers listed in the residents' telephone directory. Standardized questionnaire on social and health information, as well as a 21-item symptom checklist, were administered over the telephone. Logistic regress analyses were performed to find out if menopausal status and other social and health factors were independently associated with overall symptom reporting as well as high prevalence of reporting of each of the five symptom groups - psychological, musculoskeletal, non-specific somatic, respiratory and vasomotor. RESULTS: we observed that compared with premenopausal women, women going through menopausal transition had a 2.3- to threefold increased risk of reporting more symptoms. The prevalence of symptom reporting was lower in postmenopausal women, but remained slightly elevated compared with premenopausal women. Factors including currently not working, decreased household income in the past year, worry over friends/relatives, and health affecting daily activities were also independently associated with symptom reporting. Among these factors, reporting health affecting daily activities had the strongest association with symptom reporting. CONCLUSIONS: compared with pre and postmenopausal women, perimenopausal women had the highest reports of symptom complaints. Social, psychological and health factors were also independently associated with symptom reporting. PMID- 12590013 TI - The prevention of type 2 diabetes--lifestyle change or pharmacotherapy? A challenge for the 21st century. AB - Diabetes mellitus is occurring in epidemic proportions in many countries. In Australia 7.4% of people over 25 years of age have diabetes (mostly type 2) and comparable or higher prevalences have been reported in the United States and a number of Asian countries. The enormous economic and social cost of this disease makes a compelling case for prevention. Epidemiological studies have shown clearly that type 2 diabetes results from an interaction between a genetic predisposition and lifestyle factors including obesity, sedentary behaviour and both calorie excess and various dietary constituents. The natural history of type 2 diabetes includes a preceding period of impaired glucose tolerance (IGT)/impaired fasting glucose (IFG) which provides an opportunity for targeted intervention within large communities. Lifestyle intervention studies have consistently shown that quite modest changes can reduce the progression from IGT to diabetes by 50-60%. It may, however, not be possible to translate these successful findings to larger cohorts or maintain the lifestyle changes longer term. This has lead to consideration of pharmacotherapy. While small studies with sulphonylureas are inconclusive, benefits have been found for metformin, acarbose and troglitazone. Big intervention studies with ramipril, rosiglitazone, valsartan and nateglinide are underway. Pharmacological intervention raises a whole range of ethical, economic and practical issues not the least of which is the problem of long term therapy of the 'otherwise well'. PMID- 12590012 TI - Effects of low-dose, continuous combined estradiol and noretisterone acetate on menopausal quality of life in early postmenopausal women. AB - OBJECTIVES: To describe the effects of low dose hormonal replacement therapy (LD HRT) on quality of life in early postmenopausal women, since the postmenopausal estrogen deprivation in mid age women often brings along a series of changes and symptoms, which may greatly affect quality of life. METHODS: Fifty normal postmenopausal women were recruited and randomly treated with LD-HRT, 17beta estradiol (1 mg/day) and norethisterone acetate (0.5 mg/day) (LD-HRT) or calcium supplement (controls). No significant differences in age, age at menopause, the presence of chronic diseases and socio-economic status were present in the two groups. The Women's Health Questionnaire (WHQ), a validated quality-of-life instrument for perimenopausal and postmenopausal women, was administered at baseline and after 6 and 12 weeks of treatment in both groups. RESULTS: At baseline no significant differences in WHQ scores were present in the two groups. In the control group the scores in all different areas showed no significant modification either after 6 and 12 weeks of observation. Conversely, the LD-HRT group showed a significant decrease in the scores of vasomotor symptoms, somatic symptoms, anxiety/fear, depressed mood and sleep problem items. No effects on memory/concentration and menstrual symptoms areas were evident. CONCLUSION: Although quality of life is also and may be mainly influenced by socio-economic and cultural factors, LD-HRT definitively can improve not only vasomotor symptoms, but also more general aspects of physical and psychological well-being of symptomatic postmenopausal women. PMID- 12590014 TI - Effects of bezafibrate and pravastatin on remnant-like lipoprotein particles and lipoprotein subclasses in type 2 diabetes. AB - The effects of bezafibrate and pravastatin on remnant-like lipoprotein particles (RLPs) and lipoprotein subclasses were compared in type 2 diabetes. Bezafibrate (400 mg/day) and pravastatin (10 mg/day) were given to 27 Japanese diabetics in a randomized crossover design. RLP cholesterol (RLP-C) and RLP triglyceride (RLP TG) were measured by an immunoseparation technique. LDL and HDL were separated each into three subclasses (large, medium, small) and their cholesterol (C) contents were measured by an HPLC method. RLP-C was reduced more effectively by pravastatin (bezafibrate -16.0% vs. pravastatin -40.6%, P < 0.05), whereas RLP-TG was reduced more effectively by bezafibrate (-55.2% vs. -35.0%, P < 0.05). Further, pravastatin decreased large and small LDL-C levels equally (large; 23.6%; medium; -17.2%, small; -21.0%), while bezafibrate produced a relatively larger reduction in small LDL-C (-12.1; -16.9; -21.5%). Whereas bezafibrate significantly decreased large HDL-C and increased medium and small HDL-C (-49.6; 34.1; 35.8%), pravastatin significantly increased only medium HDL-C (5.2; 9.4; 5.9%). Bezafibrate reduced RLP-C and RLP-TG more effectively in patients with high TG levels, whereas pravastatin's effect was not markedly influenced by the initial TG level. Thus measurements of RLP-C, RLP-TG, and HPLC subclasses revealed that bezafibrate and pravastatin differently influence the lipoprotein status in type 2 diabetes. PMID- 12590015 TI - Hypoglycemia due to nateglinide administration in diabetic patient with chronic renal failure. AB - A 56-year-old woman with diabetic triopathy, rheumatoid arthritis and chronic renal failure was admitted for severe hypoglycemic coma. Arthralgia had been deteriorating for 6 months. Therefore, 5 mg of prednisolone was administered. Postprandial blood glucose (PPG), however, elevated from 260 to 290 mg/dl, although fasting blood glucose (FBG) levels ranged from 80 to 110 mg/dl. Three months after, 270 mg of nateglinide was given in addition to acarbose. After 2 days, hypoglycemia occurred at 02:00 h. Nateglinide was then decreased to 180 mg (before breakfast and lunch). After 5 days, hypoglycemia re-occurred at 01:00 h. Nateglinide was subsequently decreased to 90 mg before breakfast. The PPG levels ranged from 130 to 150 mg/dl. Hypoglycemia did not occur during the next 2 months. On admission, FBG; 59 mg/dl, fasting immunoreactive insulin; 34 microU/ml, indicated hyperinsulinemic hypoglycemia. We administered 20 g of glucose intravenously, however, hypoglycemia recurred 4 times and 20 g of glucose was then administered. Although the plasma nateglinide level decreased, the nateglinide metabolite, N-[trans-4-(1-hydroxy-1methylethyl)-cyclohexanecarbonyl] D-phenylalanine levels still had not decreased 29 h after nateglinide administration. Therefore, chronic renal failure appeared to alter the pharmacokinetic parameters of the nateglinide metabolite, which had accumulated by chronic renal failure. The nateglinide metabolite caused severe hypoglycemia in this case. PMID- 12590016 TI - Risk factors for ocular surface disorders in patients with diabetes mellitus. AB - OBJECTIVE: To evaluate risk factors for ocular surface disorders and tear dysfunction in patients with type 2 diabetes. STUDY DESIGN AND METHODS: This prospective case controlled study included 41 patients with type 2 diabetes and 20 healthy subjects as the control group. All subjects underwent routine ophthalmic examination, tear film break-up time (BUT) test, Schirmer test, fluorescein dye test, rose bengal staining test, and a questionnaire for subjective complaints. The relationship of metabolic control of diabetes mellitus, duration of diabetes, severity of diabetic retinopathy, and argon laser photocoagulation (ALP) to tear dysfunction was evaluated. RESULTS: Tear film BUT and Schirmer test values were significantly lower in diabetic patients compared with control subjects (P < 0.001). In the diabetic group, significantly more subjects had abnormal fluorescein and rose bengal staining than in the control group (P < 0.001). Abnormal tear function tests were associated with poorer metabolic glucose control, panretinal ALP, and proliferative diabetic retinopathy (P < 0.05), but not with duration of diabetes (P > 0.05). CONCLUSIONS: The results of the study indicate that poor metabolic control, panretinal ALP, and proliferative diabetic retinopathy are high risk factors for ocular surface disorders in type 2 diabetes. These patients should be followed more carefully, and should be referred to an ophthalmologist when required. PMID- 12590017 TI - Childhood diabetes identified in mass urine screening program in Taiwan, 1993 1999. AB - OBJECTIVE: To describe the gender differences in cases and characteristics of diabetes mellitus (DM) that can be identified from a mass urine screen program for school children in Taiwan. METHOD: Screening for the childhood asymptomatic proteinuria and glucosuria began in 1992 for school children. Students were instructed to collect mid-stream samples of the first morning urine for glucosuria and proteinuria tests using urine strip devices. Students with positive results for glucose and/or protein and/or occult blood in the first examination received a second urine test. The third screening test was performed for urine and fasting blood sample for 11-item examinations if the second test was positive. The 1997 criteria of American Diabetes Association were used for defining DM. RESULTS: Approximately 2,615,000-2,932,000 students received the preliminary screening each semester. The overall average rates of newly identified diabetes from 1993 to 1999 were 8.3 per 100,000 among boys, and 12.0 per 100,000 among girls. The average rate of new cases increased significantly from sixth grade for boys and fourth grade for girls, with peak rates of 14.7 per 100,000 in eighth grade for boys and 19.0 per 100,000 in sixth grade for girls. Similar prevalence trends by sex and grade were observed, higher in girls than in boys. CONCLUSIONS: This mass screening data suggest that childhood diabetes of all types in Taiwan is elevated in the age of puberty and higher in girls than in boys. PMID- 12590018 TI - Clinical features of diabetes mellitus with the mitochondrial DNA 3243 (A-G) mutation in Japanese: maternal inheritance and mitochondria-related complications. AB - Diabetes mellitus with the mitochondrial DNA 3243(A-G) mutation is reported to represent 0.5-2.8% of the general diabetic population. Since the characterization of diabetes with the mutation is still incomplete, we undertook a nation-wide case-finding study of genetically defined patients using questionnaires in Japan. One hundred and thirteen Japanese diabetic patients with the mutation were registered and analyzed. The patients had a high prevalence of maternal inheritance of diabetes and deafness, short and thin stature, and showed an early middle-aged onset of diabetes and deafness. Eighty-six percent of the patients required insulin therapy due to the progressive insulin secretory defect. Glucose intolerance of the mothers was associated with an early middle-aged onset of diabetes, reduction in the insulin secretory capacity, early requirement of insulin therapy, and increases in the daily insulin dose. The heteroplasmic concentrations of the 3243 mutation in leukocytes were low and declined with aging. The patients had advanced microvascular complications, and mitochondria related complications such as cardiomyopathy, cardiac conductance disorders, neuromuscular symptoms, neuropsychiatric disturbance, and macular pattern dystrophy. Thus, this study has revealed that: (1) diabetes mellitus with the 3243 mutation is a subtype of diabetes mellitus with mitochondria-related complications; and (2) insulin secretory ability is more severely impaired in the patients whose mothers were glucose intolerance. PMID- 12590019 TI - Association of plasma PAF acetylhydrolase gene polymorphism with IMT of carotid arteries in Japanese type 2 diabetic patients. AB - The aim of this study was to investigate association of a missense mutation in plasma PAF acetylhydrolase (G994T) with intima media thickness (IMT) of the carotid arteries. One hundred and forty Japanese type 2 diabetic patients aged from 40 to 79 years without severe nephropathy were enrolled in this study. The genotype of the patients was determined by allele specific PCR. IMT of the carotid arteries of the subjects was recorded by B-mode ultrasound imaging. The patients were divided into two groups by genotyping, one carrying two wild alleles (wild group), and another carrying one or two mutant alleles (mutant group). Each group was further divided into two subgroups according to age; one subgroup consisted of 40s or 50s, and another consisted of 60s or 70s. The prevalence of the G994T mutation in the subjects was 28.6% (24.3% heterozygote, and 4.3% homozygote). IMT of the elderly patients of the mutant group was significantly greater (0.98 +/- 0.22 mm, n = 26) than of the elderly patients of the wild group (0.87 +/- 0.20 mm, n = 50, P = 0.0292). There was no significant difference in clinical characteristics between the two subgroups. The results of this study indicate that the missense mutation in plasma PAF acetylhydrolase is associated with development of atherosclerosis in the elderly. PMID- 12590020 TI - Relative role of insulin resistance and beta-cell dysfunction in the progression to type 2 diabetes--The Kinmen Study. AB - This study compared the relative role of insulin resistance and beta-cell dysfunction (both assessed using the HOMA method) with glucose intolerance conditions in the progression to type 2 diabetes among a high risk group of subjects with fasting plasma glucose (FPG) 5.6-7.0 mmol/l in Kinmen, Taiwan. Data were collected during a continuing prospective study (1998-99) of a group of Taiwanese subjects at high-risk of developing type 2 diabetes who had fasting hyperglycemia (5.6-7.0 mmol/l) and exhibited 2-h postload glucose concentrations <11.1 mmol/l from 1992-94 to 1995-96. Among 644 non-diabetic subjects at baseline, 79.8% (514/644) had at least one follow-up examination. There were 107 new cases of diabetes diagnosed by 1999 WHO criteria in 2918.7 person-years of follow-up. The incidence rate was 3.67%/year (107/2918.7). After adjustment for other possible associative variables, including gender, age, BMI, waist circumference, insulin resistance, and beta-cell dysfunction, Cox's hazard model showed that those individuals with isolated IFG (impaired fasting glucose) and those individuals with isolated IGT (2-h glucose impairment) exhibited similar risk of developing diabetes. Those individuals with isolated IFG and isolated IGT showed a comparable impairment of basal or hepatic insulin sensitivity, but those individuals with isolated IFG had a greater beta-cell dysfunction by the HOMA method. PMID- 12590021 TI - Alternative site testing increases compliance of SMBG (preliminary study of 3 years cohort trials). PMID- 12590023 TI - Myocardial infarction associated with diffuse pain. PMID- 12590022 TI - The foundation of a medical faculty at the University of Namibia. PMID- 12590025 TI - Ethical validity of palliative sedation therapy. PMID- 12590026 TI - Validation study of the Japanese version of the brief fatigue inventory. AB - Fatigue has been recognized as one of the most distressing symptoms in cancer patients. Concise assessment is essential to managing this symptom. To that end, the Brief Fatigue Inventory (BFI), a 9-item questionnaire, was designed to assess fatigue in cancer patients. The purpose of this study was to examine the validity and reliability of the Japanese version of this scale (BFI-J), when compared with previously validated fatigue instruments. We randomly selected 252 cancer patients and presented them with the BFI-J, along with the Cancer Fatigue Scale; Profile of Mood States fatigue, vigor, and depression subscales; and European Organization for Research and Treatment of Cancer QLQ-C30. Specifically, the reliability and construct, criterion, convergent, and discriminant validity of each instrument were evaluated. Additionally, fatigue severity classification was explored using the BFI-J. The results indicated that the BFI-J is a brief, valid, and feasible measure of fatigue for use with Japanese cancer patients. PMID- 12590024 TI - Barriers to pain and symptom management, opioids, health policy, and drug benefits. PMID- 12590027 TI - Factor analysis of laboratory and clinical measurements of dyspnea in patients with chronic obstructive pulmonary disease. AB - The primary purpose of this study was to determine if there are three distinct factors representing ratings of dyspnea during laboratory exercise, clinical ratings of dyspnea, and pulmonary function in patients with moderate to severe chronic obstructive pulmonary disease (COPD) (n = 92) using factor analysis. Subjects (mean age 66 +/- 7 yrs; FEV1% predicted 44.7 +/- 14.0) were randomly assigned to one of three treatment groups to test the effects of three education and exercise training programs. Outcomes were evaluated at baseline and at 2 months after the intervention. Dyspnea ratings with laboratory exercise (SOB) were measured during incremental (ITT) and endurance (ETT) treadmill tests, and a six-minute walk (6MW) using the modified Borg scale. Clinical measures of dyspnea were measured with the Baseline and Transitional Dyspnea Index (BDI/TDI), UCSD Shortness of Breath Questionnaire (SOBQ), Medical Research Council Dyspnea Scale (MRC), Dyspnea subscale of the Chronic Respiratory Questionnaire (CRQ-D), and a global dyspnea question. Pulmonary function parameters included FEV1% predicted, FEV1/FVC, and RV/TLC. The factor analysis yielded three factors that accounted for 58.7% of the total variance in the data: Factor 1, "Dyspnea with Laboratory Exercise" comprised ETT SOB end, ETT SOB isotime, ITTSOB end, ITT SOB isotime, ITT SOB/Time and ETT SOB/Time. Factor 2, "Clinical Dyspnea," comprised 6MW SOB, 6MW SOB/Feet, BDI, SOBQ, MRC, Global SOB, CRQ-D. Measures of airway resistance (FEV1% predicted, FEV1/FVC) and hyperinflation (RV/TLC) loaded on a third factor, "Pulmonary Function." An additional post hoc factor analysis with post intervention data provided similar results. The Global SOB question and ITT SOB isostage variables were relatively more sensitive to change compared to the other outcome variables. We conclude that pulmonary function, clinical ratings of dyspnea, and laboratory ratings of dyspnea are three separate and independent factors and should be included in the routine clinical evaluation of patients with COPD. PMID- 12590028 TI - Surveys to assess satisfaction with end-of-life care: does timing matter? AB - The goals of this study were to determine whether post-death surveys of family members cause more distress if they are administered closer in time to the patient's death, and whether family members are less likely to respond to earlier surveys. Caregivers of hospice patients were randomly assigned to receive a survey at 2 weeks (n = 107) or at 6 weeks (n = 100) after the patient's death. Response rates and self-reported distress experienced in completing the survey were recorded. There were no differences in self-ratings of distress between 2- and 6-week surveys, and response rates were identical (2-week: 54%; 6-week: 54%). Distress and response rate do not appear to be influenced by the timing of data collection, even when surveys are administered very soon after death. PMID- 12590029 TI - The COX-2 specific inhibitor, valdecoxib, is an effective, opioid-sparing analgesic in patients undergoing total knee arthroplasty. AB - This multicenter, randomized, double-blind, placebo-controlled study evaluated the analgesic efficacy and opioid-sparing effects of valdecoxib, a potent COX-2 specific inhibitor, in patients undergoing knee replacement. Patients received morphine by patient-controlled analgesia (PCA), and valdecoxib 40 mg or 80 mg daily, or placebo, for up to two days. Efficacy was assessed by the cumulative amount of morphine administered over 48 hours, pain intensity and patient's evaluation of medication. Morphine consumption over 48 hours by patients receiving valdecoxib 40 mg or 80 mg daily plus morphine was 83.7% and 75.8% (P < 0.05) of the total amount consumed by patients receiving morphine alone. Patients receiving valdecoxib 40 mg and 80 mg daily experienced significantly lower maximum pain intensity on Day 2 (P < 0.05), and rated their study medication significantly higher than patients receiving morphine alone. Valdecoxib plus morphine was well tolerated. Thus, valdecoxib in combination with morphine provides multi-modal analgesia that reduces pain and opioid use and increases patient satisfaction following knee replacement surgery. PMID- 12590030 TI - A comparison of the pharmacokinetics, clinical efficacy, and tolerability of once daily tramadol tablets with normal release tramadol capsules. AB - This report describes a pharmacokinetic study and a clinical study of three different formulations of oral tramadol: once-daily tramadol tablets 150 and 200 mg, and normal release tramadol capsules 50 mg 8-hourly. The randomized, open label, crossover pharmacokinetic study included 22 subjects. The three treatments showed equivalent mean systemic availability of tramadol. The mean relative systemic availabilities (90% confidence intervals) for the once-daily tablets 150 mg and the once-daily tablets 200 mg versus the normal release capsules were 89.6 (83.3-95.8)% and 90.5 (84.6-96.8)%, respectively. The values for the once-daily tablet 150 mg versus the once-daily tablet 200 mg were 99.0 (92.6-105.9)%. The randomized, double-blind, double-dummy, crossover clinical study included 134 patients with moderate osteoarthritic pain. The three treatments showed similar efficacy; they all reduced patients' pain scores from baseline and there were no significant treatment differences in pain scores during treatment. The three treatments were also well tolerated. PMID- 12590031 TI - Is there evidence that palliative care teams alter end-of-life experiences of patients and their caregivers? AB - Palliative care provision varies widely, and the effectiveness of palliative and hospice care teams (PCHCT) is unproven. To determine the effect of PCHCT, 10 electronic databases (to 2000), 4 relevant journals, associated reference lists, and the grey literature were searched. All PCHCT evaluations were included. Anecdotal and case reports were excluded. Forty-four studies evaluated PCHCT provision. Teams were home care (22), hospital-based (9), combined home/hospital care (4), inpatient units (3), and integrated teams (6). Studies were mostly Grade II or III quality. Funnel plots indicated slight publication bias. Meta regression (26 studies) found slight positive effect, of approximately 0.1, of PCHCTs on patient outcomes, independent of team make-up, patient diagnosis, country, or study design. Meta-analysis (19 studies) demonstrated small benefit on patients' pain (odds ratio [OR]: 0.38, 95% confidence interval [CI]: 0.23 0.64), other symptoms (OR: 0.51, CI: 0.30-0.88), and a non-significant trend towards benefits for satisfaction, and therapeutic interventions. Data regarding home deaths were equivocal. Meta-synthesis (all studies) found wide variations in the type of service delivered by each team; there was no discernible difference in outcomes between city, urban, and rural areas. Evidence of benefit was strongest for home care. Only one study provided full economic cost-benefit evaluation. This is the first study to quantitatively demonstrate benefit from PCHCTs. Such comparisons were limited by the quality of the research. PMID- 12590032 TI - A systematic review of hydromorphone in acute and chronic pain. AB - While morphine is historically the gold standard for the management of severe cancer pain, some patients either do not achieve adequate analgesia, or suffer intolerable side effects from this drug. For these patients, alternatives such as hydromorphone are recommended. This review explores the evidence for the efficacy of hydromorphone in the management of pain. A systematic search, from 1966 to 2000, of published and unpublished randomized trials that involved the administration of hydromorphone for both acute and chronic pain conditions in adults and children, was conducted. Forty-three studies were included in the review; 11 involved chronic cancer pain and 32 acute pain. Approximately half the studies received a low quality score. In addition, the heterogeneity of the studies precluded combination of data and results. Overall, hydromorphone appears to be a potent analgesic. The limited number of studies available suggests that there is little difference between hydromorphone and other opioids in terms of analgesic efficacy, adverse effect profile and patient preference. However, most studies involved small numbers of patients and wide ranges in equianalgesic dose ratios, making it difficult to determine real differences between interventions. PMID- 12590033 TI - Is there still a role for open cordotomy in cancer pain management? AB - For a small number of cancer patients, good pain control remains difficult to achieve despite adequate assessment and medical management. In nine cases, effective control of intractable pain from malignant pelvic disease was achieved by open thoracic cordotomy. The technique was well tolerated, with no major complications. Eight of the nine patients decreased their median daily oral morphine requirement from 560 mg (range 360-2600 mg) to 160 mg (range 40-1000 mg). Maximal survival time post-cordotomy was 830 days, with a median of 107 days. No patient experienced recurrent pain in the initially painful site. For patients with intractable pain associated with advanced pelvic malignancy, the use of an open cordotomy should be considered when satisfactory pain control is not achieved by medical or minimally invasive methods. PMID- 12590034 TI - Hypocalcemia following pamidronate administration for bone metastases of solid tumor: three clinical case reports. AB - Bisphosphonates, such as pamidronate, are a new class of drugs, initially described for treatment of neoplasic hypercalcemia. Currently, they also may be used in the treatment of bone metastases from solid tumor, even without hypercalcemia. Hypocalcemia is a potential adverse effect of these drugs, which is considered infrequent and rarely symptomatic. We describe three cases of severe hypocalcemia following one injection of pamidronate. The three patients had bone metastases from solid tumors (breast in two cases, prostate in one), at least partially osteoblastic, and none had hypercalcemia. The induced hypocalcemia was rapid in onset, severe, and durable. The mechanism seems to be multiple and may include both the expected reduction of osteolysis and also a rapid and direct action on parathyroid glands followed by resistance to parathormone. Some elements could amplify the phenomenon, such as latent hypoparathyroidism after surgery, cervical radiotherapy, hypomagnesemia, or low 25 hydroxy vitamin D (25OH D). For patients who have such risk factors, it may be useful to check calcium several days after the first injection. PMID- 12590035 TI - Case presentation: medical decision-making in hopeless situations: the long-lost son. PMID- 12590036 TI - Miracles and medicine: helping families navigate the course. PMID- 12590038 TI - Surprises: low probabilities or high contrasts? AB - Outcome expectations can be expressed prospectively in terms of probability estimates, and retrospectively in terms of surprise. Surprise ratings and probability estimates differ, however, in some important ways. Surprises are generally created by low-probability outcomes, yet, as shown by several experiments, not all low-probability outcomes are equally surprising. To account for surprise, we propose a contrast hypothesis according to which the level of surprise associated with an outcome is mainly determined by the extent to which it contrasts with the default, expected alternative. Three ways by which contrasts can be established are explored: contrasts due to relative probabilities, where the obtained outcome is less likely than a default alternative; contrasts formed by novelty and change, where a contrast exists between the obtained outcome and the individual's previous experience; and contrasts due to the perceptual or conceptual distance between the expected and the obtained. In all these cases, greater contrast was accompanied by higher ratings of surprise. PMID- 12590037 TI - Beyond ethnicity. PMID- 12590039 TI - Six-year-olds' difficulties handling intensional contexts. AB - Previous research shows that children have difficulties handling intensional contexts even when they can pass a test of false belief (e.g. Cognition 67 (1998) 287; Cognition 25 (1987) 289). Some authors (Perner, J. (1991). Understanding the representational mind. Cambridge, MA: MIT Press; Cognition 25 (1987) 289) place these difficulties in the linguistic and not the mental representational domain. The experiments reported here examined whether 6-year-old children could answer questions in an intensional context that did not require the explicit verbal characterization of a belief. We replicated previous findings and found that children answered according to their own knowledge in an intensional context. This occurred even though they responded by choosing a picture to insert into a protagonist's thought bubble rather than report the belief verbally. Children could correctly answer questions about the knowledge state of the protagonist and pass a test of false belief. Further experiments ruled out methodological explanations. Experiment 2 showed that the difference in answering according to own knowledge between the false belief and intensional stories is not accounted for by procedural factors in the two types of test. Experiment 3 revealed that children did not answer according to their own knowledge by default. Experiment 4 suggested that answering according to own knowledge was not a result of pictorial salience. Results are discussed in relation to the simulation-theory debate. PMID- 12590040 TI - On the processing of regular and irregular forms of verbs and nouns: evidence from neuropsychology. AB - Following acquired brain damage, a native English speaking patient (AW) encountered problems accessing phonology in speech production, while her ability to access word meaning appeared to be intact. In a series of tasks, AW was presented either with a verb, and was asked to produce its past tense or past participle (walk --> "walked"), or with a noun, and was asked to produce its plural (glove --> "gloves"). A stark dissociation was found: while AW responded accurately with regular forms of verbs (walked) and nouns (gloves), performance was significantly less accurate with irregular forms (found; children). The appearance of a selective deficit for irregular forms in conditions of impaired lexical access is in line with dual-mechanism accounts, which proposes that irregular forms are specified in the lexicon whereas regular forms are computed via rule-based mechanisms. In contrast, AW's data are problematic for connectionist accounts that do not posit separate mechanisms for processing regular and irregular forms, including the connectionist model recently proposed by Joanisse and Seidenberg (Proceedings of the National Academy of Sciences USA 96 (1999) 7592) which successfully simulated a variety of earlier neuropsychological findings. Analyses of AW's responses shed light on further details of the representation and processing of regular and irregular inflected forms. PMID- 12590041 TI - Alphabetic and nonalphabetic L1 effects in English word identification: a comparison of Korean and Chinese English L2 learners. AB - Different writing systems in the world select different units of spoken language for mapping. Do these writing system differences influence how first language (L1) literacy experiences affect cognitive processes in learning to read a second language (L2)? Two groups of college students who were learning to read English as a second language (ESL) were examined for their relative reliance on phonological and orthographic processing in English word identification: Korean students with an alphabetic L1 literacy background, and Chinese students with a nonalphabetic L1 literacy background. In a semantic category judgment task, Korean ESL learners made more false positive errors in judging stimuli that were homophones to category exemplars than they did in judging spelling controls. However, there were no significant differences in responses to stimuli in these two conditions for Chinese ESL learners. Chinese ESL learners, on the other hand, made more accurate responses to stimuli that were less similar in spelling to category exemplars than those that were more similar. Chinese ESL learners may rely less on phonological information and more on orthographic information in identifying English words than their Korean counterparts. Further evidence supporting this argument came from a phoneme deletion task in which Chinese subjects performed more poorly overall than their Korean counterparts and made more errors that were phonologically incorrect but orthographically acceptable. We suggest that cross-writing system differences in L1s and L1 reading skills transfer could be responsible for these ESL performance differences. PMID- 12590042 TI - Changing perspective within and across environments. AB - Perspective change within a single environment is a slow and effortful process. However, little research has addressed perspective change across multiple environments. Using a task-set switching paradigm, subjects judged spatial relationships between target locations from differing perspectives. Response times were longer when successive trials probed different perspectives. However, this cost was greater when perspective was changed within a single environment compared to when it was changed across two environments. This result indicates that the processing of perspective change, and perhaps general spatial reasoning, differs in these two cases. Implications for theories of perspective change and environmental knowledge are discussed. PMID- 12590043 TI - Infants learn phonotactic regularities from brief auditory experience. AB - Two experiments investigated whether novel phonotactic regularities, not present in English, could be acquired by 16.5-month-old infants from brief auditory experience. Subjects listened to consonant-vowel-consonant syllables in which particular consonants were artificially restricted to either initial or final position (e.g. /baep/ not /paeb/). In a later head-turn preference test, infants listened longer to new syllables that violated the experimental phonotactic constraints than to new syllables that honored them. Thus, infants rapidly learned phonotactic regularities from brief auditory experience and extended them to unstudied syllables, documenting the sensitivity of the infant's language processing system to abstractions over linguistic experience. PMID- 12590044 TI - Allele sharing in first-degree and unrelated pairs of individuals in the Ge F I AmpFlSTR Profiler Plus database. AB - Eleven Italian forensic laboratories participated in a population study based on the AB Profiler Plus loci with proficiency testing. The validated database, including 1340 individuals, is available on-line. Tests for Hardy-Weinberg equilibrium, gametic unbalance, and heterogeneity of gene frequency were generally not significant. Gene frequencies at each locus were consistent with those of two previously published Italian studies, but different from a third. Individuals of each subsample were paired, and the total number of alleles shared across the nine loci was determined in each pair. The analysis was replicated over the total sample. In addition, two samples of mother-child pairs (N=315) and full-sib pairs (N=91) were subjected to allele sharing analysis. The resulting distributions were sufficiently distinct from the sample of unrelated pairs as to be of practical usefulness. PMID- 12590045 TI - Effect of hair care and hair cosmetics on the concentrations of fatty acid ethyl esters in hair as markers of chronically elevated alcohol consumption. AB - Fatty acid ethyl esters (FAEE) can be used as alcohol markers in hair. It was investigated in this study whether this diagnostic method is disturbed by hair care and hair cosmetics. Traces of ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate were detected in all of 49 frequently applied hair care products by headspace solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). The highest concentration was 0.003% in a hair wax. From experiments with separated hair samples of alcoholics as well as from the evaluation of the FAEE concentrations and the data about hair care of 75 volunteers (alcoholics, social drinkers and teetotalers) follows that usual shampooing, permanent wave, dyeing, bleaching or shading are of minor importance as compared to the drinking amount and other individual features. However, false positive results were found after daily treatment with a hair lotion containing 62.5% ethanol, with a deodorant and with a hair spray. As an explanation, it is assumed that FAEE are formed in the sebum glands also after regular topical application of products with a higher ethanol content. PMID- 12590046 TI - Determination of drugs of abuse in hair: evaluation of external heroin contamination and risk of false positives. AB - One of the most controversial point regarding the validity of hair testing is the risk of false positive due to external contamination. The aim of our experience is to verify if a 5 consecutive days contamination with a small amount of a powdered mixture of heroin hydrochloride and acetylcodeine hydrochloride (10:1 w/w) will last sufficiently long to make a contaminated subject indistinguishable from active users, and if normal washing practices together with the decontamination procedure are sufficient to completely remove the external contamination. Our results suggest that decontamination procedures are not sufficient to remove drugs penetrated into hair from external source. In fact, all contaminated subjects were positive for opiates (heroin, 6-MAM, morphine, acetylcodeine and codeine) for at least 3 months. Significant 6-MAM concentrations (>0.5 ng/mg) were found in each subject until 6th week. Further, 6 MAM/morphine ratio were always above 1.3. PMID- 12590047 TI - Suicide by injection of a veterinarian barbiturate euthanasia agent: report of a case and toxicological analysis. AB - The history and toxicological findings of a suicidal case involving injection of a veterinarian barbiturate euthanasia agent (Vetanarcol containing pentobarbital are presented. Blood pentobarbital concentrations compatible with drug overdose were determined. Almost identical levels were found in blood, cerebrospinal fluid (CSF) and vitreous humour (VH). The highest concentration was measured in the bile. The present case is compared with similar rare cases in the literature. PMID- 12590048 TI - Tissue distribution of nitrazepam and 7-aminonitrazepam in a case of nitrazepam intoxication. AB - We report a case of nitrazepam poisoning in which the distribution of nitrazepam and 7-aminonitrazepam was determined in body fluids and tissues. A 52-year-old woman was found dead in a shallow ditch (approximately 5 cm in depth), in the winter. Ambient temperature was 2-8 degrees C. The postmortem interval was estimated to be approximately 1 day and no putrefaction was observed. The cause of death was thought to be drowning due to nitrazepam overdose and cold exposure. Blood concentrations of nitrazepam and 7-aminonitrazepam were very site dependent (0.400-0.973 microg/ml and 0.418-1.82 microg/ml). In addition, the concentration of the same analytes in the bile were 4.08 and 1.67 microg/ml, respectively, and in the urine: 0.580 and 1.09 microg/ml, respectively. A high accumulation of both substances was observed in various types of brain tissue (2.17-6.22 microg/g and 2.49-5.11 microg/g). Only small amounts of nitrazepam and 7-aminonitrazepam were detected in the liver (0.059 and 0.113 microg/g, respectively). Large differences in the observed concentrations of nitrazepam and 7-aminonitrazepam among arterial and venous blood samples were thought to be mainly due to dilution of arterial blood by water entering the circulation through lungs at the time of death. Bacterial metabolism of nitrazepam may also have contributed to the observed differences. PMID- 12590049 TI - Cause-of-death query in validation of death certification by expert panel; effects on mortality statistics in Finland, 1995. AB - The correctness of selection, coding and registration of underlying cause-of death is important for the quality of mortality statistics. One measure to improve quality is the query to the certifier for verification of the underlying cause-of-death. In Finland, 3478 death certificates, 7.1% of total 49074 certifications in 1995, were considered questionable by statisticians. The expert panel at Statistics Finland was able to resolve 2813 (80.9%) of them. However, 665 (19.1%) certificates needed to be further queried from the certifier. Of these, 318 (47.8%) were re-assigned to another ICD-9 category or to the applicable three-digit category within the main category of heart and vascular diseases, resulting in changes from a 17.00-fold increase in rheumatic heart diseases (ICD-9 codes 390-398) to a decrease of about one-half (0.45-fold change) in unspecified neoplasms (codes 235-239). However, a statistically significant impact on national mortality statistics was not observed in any of applied ICD categories. Among all questionable death certificates, most prone to query of the certifier, and with a statistical significance of P<0.05, were those with no cause-of-death specified, those stating underlying cause-of-death as non specified neoplasms (with a observed/expected ratio, O/E, of 1.69), and heart and vascular diseases (1.45), with its subcategories of ischaemic heart diseases (1.33) and other heart diseases (2.92). Death certificate validation, by expert panel consultations and query to the certifiers, and the importance of estimation of the validity of cause-of-death information on death certificates are strongly pointed out in a continuous strive for correct and reliable mortality statistics. PMID- 12590050 TI - Analysis of 418 cases of sexual assault. AB - The aim of the study was to describe victim, assailant, assault characteristics for sexual assault victims according to the time between the last sexual assault and the examination, and to provide descriptive data on medico-legal findings. The study was based on 418 examined victims of sexual assault during the year 1998. Victims were referred from investigating police authorities. All examinations were performed with the use of colposcopy by physicians with special training in forensic medicine. Two groups of victims were defined:a first group of victims examined in emergency within 72 h after the last sexual assault;a second group of victims examined after 72 h. About 86% of the cases were female victims. The mean age of the cases of the first group was 22.4 years. Conversely, 76% of the victims examined after 72 h were under the age of 15 years. Vulnerability was present in 31% of the cases examined in emergency, including disabled and pregnant victims. Sexual assault happened once in 87% of the cases of the first group and in 64% of the cases of the second group. The victim's home was the most frequent place of sexual assault (35% of the cases of the first group and 56% of the cases of the second group). The assailant was a stranger in 51% of the cases of the first group. In the second group of the victims, the assailant was a family member in 58% of the cases (the father in 30% of the cases). There was a single assailant in the majority of the cases for the two groups. Threats were used by the assailant in 66% of the victims examined in emergency and in 33% of the cases of the second group. The type of sexual assault was penetration in the majority of the cases for the two groups. Vaginal, oral and anal penetration was respectively involved in 55, 23 and 13% of the cases of the first group. General body trauma was found in 39.1% of the cases examined in emergency and in 6.3% of the cases of the second group. Genital trauma occurred in 35.7% of the cases of the first group and in 19.5% of the cases of the second group. Hymenal, vulvo-vaginal and anal lesions were respectively found in 11, 20 and 7% of the cases examined in emergency. Toxicological analysis was performed in 14.3% of the cases examined in emergency. In 47% of the tested cases, drug was detected. Cytology was performed in 61.5% of the cases examined in emergency. Detection of spermatozoa was found in 30.3% of these cases. Our study has shown that sexual assault victims had different characteristics according to the time between the sexual assault and the examination. Public health campaigns against sexual abuse and rape as well as medical management of the sexually assaulted victims should adapt to the needs and the characteristics of these two different populations of victims. PMID- 12590051 TI - Frequency of bone/cartilage lesions in stab and incised wounds fatalities. AB - A retrospective study was carried out on 58 fatalities due to stab or incised wounds. The frequency of bone or cartilage lesions was analysed according to the number of wounds, the circumstances of death and the anatomical site. Our findings showed that bone/cartilage lesions were present in about 53% of the cases. Cartilage lesions were more frequent than bone lesions. The mean number of wounds in the group with bone/cartilage lesions was statistically higher than the mean number of wounds in the group without lesions (P=0.0068). The main cause of death was thoracic injury in the groups with and without bone/cartilage lesions. In case of skeletal remains, only bone or cartilage lesions allow to diagnose stab or incised wounds. The discovery of these lesions, often of small size, justifies a complete and careful examination of skeletal remains with the help of stereomicroscopy. PMID- 12590052 TI - Mitochondrial DNA cytochrome oxidase I gene: potential for distinction between immature stages of some forensically important fly species (Diptera) in western Australia. AB - Forensic entomology requires the fast and accurate identification of insects collected from a corpse for estimation of the postmortem interval (PMI). Identification of specimens is traditionally performed using morphological features of the insect. Morphological identification may be complicated however by the numerical diversity of species and physical similarity between different species, particularly in immature stages. In this study, sequencing was performed to study the mitochondrial DNA (mtDNA) as the prospective basis of a diagnostic technique. The sequencing focused on a section of the cytochrome oxidase I encoding region of mtDNA. Three species of calliphorid (blow flies) commonly associated with corpses in western Australia, Calliphora dubia, Chrysomya rufifacies and Lucilia sericata, in addition to specimens of Calliphora augur and Chrysomya megacephala were studied. Phylogenetic analysis of data revealed grouping of species according to genus. The DNA region sequenced allowed identification of all species, providing high support for separation on congeneric species. Low levels of variation between some species of the same genus however indicate that further sequencing is required to locate a region for development of a molecular-based technique for identification. PMID- 12590053 TI - Post-mortem changes in calmodulin binding proteins in muscle and lung. AB - Estimation of post-mortem interval (PMI) remains an elusive issue in forensic investigations. In this study, we examined the possible use of calmodulin (CaM) binding proteins (CaMBPs) as indicators of PMI. Whole CaMBP populations from homogenized rat lung and rat skeletal muscle removed at 0, 24, 48 and 96 h post mortem at 21 degrees C were detected by the calmodulin binding overlay technique (CaMBOT) using 35S-VU1-CaM and visualized by autoradiography. CaMBOT showed that, in both tissues, the CaMBP population remained relatively stable for up to 96 h post-mortem with the exception of a single approximately 200 kDa CaMBP that increased in 24 h post-mortem samples then showed decreasing amounts at subsequent times. Immunoblot analysis of the specific CaMBPs, Ca(2+)/CaM dependent kinase II (CaMKII), calcineurin A (CNA), myristoylated alanine-rich C kinase substrate (MARCKS) and inducible nitric oxide synthase (iNOS) were done on lung tissue samples. CaMKII levels did not change appreciably over the 96 h PMI examined. In contrast to iNOS levels, which varied from sample to sample, CNA and MARCKS showed predictable patterns of change: the level of MARCKS decreased steadily in the 0-96 h post-mortem lung samples while CNA underwent a shift in mobility on SDS-PAGE by 24 h post-mortem before slowly decreasing in amount. The stability of CaMKII levels over 96 h was also seen in skeletal muscle tissue while CNA showed variable levels at 0, 48 and 96 h with the presence of the rapidly migrating band at 24 h. These patterns of change in CaMBPs provide some insight into the post-mortem changes in calmodulin-mediated signaling components in lung and skeletal muscle and support the further study of CNA and CaMKII as potential markers for estimating short- and long-term PMIs. PMID- 12590054 TI - Performance of immunoassays in screening for opiates, cannabinoids and amphetamines in post-mortem blood. AB - Several immunoassay methods for screening of abused drugs in whole blood were evaluated in post-mortem forensic toxicology. Blood samples known to be positive or negative for opiates, cannabinoids or amphetamines by gas chromatography-mass spectrometry (GC-MS) were analysed by EMIT II Plus and EMIT d.a.u., Syva RapidTest and Triage 8 after acetone precipitation. In these experiments, the EMIT immunoassay method was modified by using the Dade Behring VIVA analyser to detect substances more sensitively. Low concentrations of abused drugs were detected in blood samples. The sensitivities of the modified EMIT method for opiates, cannabinoids and amphetamines were 100, 86 and 98%, respectively, whereas the values were below 86% with the other methods. The specificities of all immunoassay methods for opiates and cannabinoids were 83% or above but 51-85% for amphetamines. Sample rejection occurred in a few cases with the EMIT amphetamine assays. The modified EMIT immunoassay system presented here seems to be useful for screening of drugs of abuse in post-mortem blood samples, especially when urine is not available. PMID- 12590055 TI - The time-dependent expression of fibronectin, MRP8, MRP14 and defensin in surgically treated human skin wounds. AB - Fibronectin, MRP8, MRP14 and defensin were detected immunohistochemically in 46 surgically treated, fresh (hours old) human wounds and in wounds of 13 individuals who died immediately from fatal trauma (airplane crashes or train rollovers). In immediate fatal trauma, it was not possible to detect fibronectin outside of bleeding areas nor could inflammatory cells be visualized in the interstitium using MRP8, MRP14, or defensin antibodies. Fibronectin staining could be regularly demonstrated in wounds at least 20 min-old. Granulocytic infiltrates limited to the perivasal space could be detected 20-30 min after infliction of the wound expressing MRP8, MRP14 and defensin. It was also possible to detect fibronectin networks and MRP8-, MRP14-, and defensin-positive granulocytes and macrophages in particular wounds up to 30 days-old. No differences between the expression of MRP8 and MRP14 could be demonstrated in the wounds, the majority of which were only several hours old. As wound age increased, the number of defensin-positive granulocytes detected decreased. The immunohistochemical detection of fibronectin is a useful way to demonstrate vitality in fresh wounds, beginning about 20-30 min post-trauma. However, detection of MRP8, MRP14 and defensin provides no advantage over the routine histological detection of granulocytes and macrophages in wounds under 1-2 days old. PMID- 12590057 TI - Evaluation of links in heroin seizures. AB - The evaluation of a link between two heroin seizures using a descriptive method is presented. It is based on the measure of the angles between two chromatograms assimilated to vectors, and interpreted using a continuous approach based on the likelihood ratio of Bayes' theorem. A complete evaluation model thus avoids the drawbacks of decision thresholds used until now to establish a link. Validation is obtained through tests and simulation methods. PMID- 12590056 TI - Narcotics at street level in Denmark. A prospective investigation from 1995 to 2000. AB - This article describes an investigation of illicit drugs at street level in six selected police districts in different regions of Denmark. The investigation was carried out during a 6-year period from 1995 to 2000. During the period, a total of 1244 samples were examined, as about 200 samples were seized each year. A total of 94% of the seized samples were familiar drugs: heroin base, heroin hydrochloride, cocaine hydrochloride and amphetamine sulphate. Only 2% of the samples contained designer drugs. From having constituted 53% of the samples in 1995, the frequency of heroin base fell during the period to 27% of the samples in 2000. The frequency of heroin hydrochloride was unchanged. In the same period, the frequency of cocaine hydrochloride increased from 10% of the samples in 1995 to about 25% of the samples in 2000. Apart from a few exceptions, cocaine had the same extension in all regions of Denmark after 1996. Amphetamine was more frequent in the west of Denmark, while heroin hydrochloride was more frequent in central Denmark. The purity of heroin base was lower in the period 1997-1999 than in the other years. During the entire period, the purity of cocaine hydrochloride and amphetamine sulphate fell, while the purity of heroin hydrochloride was unchanged. No significant differences between the various regions of Denmark were detected during the period in the purity of heroin hydrochloride, heroin base, cocaine hydrochloride or amphetamine sulphate. MDMA was the most frequent designer drug, but other types of designer drugs such as MDA and MDE and the less common PMA, PMMA and MBDB were also found. PMID- 12590058 TI - A comparison of adjustment methods to test the robustness of an STR DNA database comprised of 24 European populations. AB - An aim of the European Network of Forensic Science Institutes (ENFSI) is to produce a DNA database of second generation multiplex (SGM) STR profiles that is representative of the resident cosmopolitan populations. To achieve this, data were collected from 24 different populations. All of the data were combined to form one database of 5700 profiles from which allele proportions were calculated. The robustness of this combined European database was tested by estimating parameter d for every DNA profile, where d=log(10)(Pm(c)/Pm(E)) Pm(c) is the match probability of the profile calculated from its cognate database and Pm(E) is the match probability of the combined European database. Overall there was a small tendency for Pm(c)>Pm(E) primarily because of sampling bias. This bias was removed by the simple expediency of applying an adjustment factor to the calculation of Pm(E). These were selected from the Balding size bias correction, the Balding and Nichols Fst correction, a minimum allele proportion (between 0.01 and 0.02), an upper bound of a 95% confidence interval (CI) and a lower bound on the genotype match probability. It was demonstrated that a single European database is a feasible proposition. A combination of different adjustment methods can be used to ensure that the result is conservative relative to the cognate database, and their effect measured by parameter d. PMID- 12590059 TI - Complex suicides by self-incineration. AB - Among the few cases of suicidal self-incineration primary or secondary complex suicides are a rarity. Accordingly the number of reports on this subject in the forensic literature is small. In a retrospective analysis of the Freiburg and Berlin autopsy material, four cases were found within an observation period of 11 years. These are described together with seven other cases reported in the literature. Altogether most of these suicides are primary combinations, especially with falls from a height. Other combinations include hanging or the infliction of stabs, cuts or gunshot injuries. The sex and age distribution is similar to that found for suicide by self-incineration alone. PMID- 12590060 TI - Preliminary study of hair form of Japanese head hairs using image analysis. AB - The use of average curvature measurements for the forensic comparison of curly hairs has been reported, but a method, in which various types of hair form are quantitatively examined and objectively interpreted for hair comparison, has not been reported to date. In the present study, numerical data on hair form from Japanese subjects were obtained by image analysis and a morphological comparison of these head hairs was investigated. Head hairs obtained from eight Japanese males were measured for length (L), distance (D) and area (A) using a Kontron Imaging System KS400. From the three measurements mentioned above, three indexes, L/D, A/D and 2(A/L), were examined. The inter-individual variations for each value were investigated by a t-test and the availability of six values for the forensic comparison of hair form was evaluated by a stepwise linear discrimination analysis. Six values obtained from hair form by an image analysis showed large intra-individual variations. However, these six values were found to be useful for discriminating between two individuals, since the six values showed larger inter-individual variations than intra-individual variations. Discrimination on each comparison using a stepwise linear discrimination analysis was performed for some of the values and the results indicated conspicuous inter individual variations between the two individuals. On 11 of 28 comparisons, 30 hairs from one individual could be completely distinguished from hairs of another individual, when a two-way comparison was employed. These results confirm that hair form could be quite useful in the forensic comparison of hair morphology, and suggest that numerical data obtained from hair form by image analysis are very important values for constructing a screening procedure for evidential hairs. The use of an objective measure of hair form will be especially useful for Japanese head hairs since they are generally thought to show very limited variation in morphological features. PMID- 12590061 TI - Population data of nine Y-chromosomal STR loci in northern Poland. AB - Allele and haplotype frequencies of nine Y-chromosome STR loci were determined from a sample of 508 unrelated males from north Poland. Three samples containing insertion polymorphism DYS19 (duplication) and DYS385I/II (triplications) were observed. PMID- 12590062 TI - STRs data for the loci D3S1385, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820 from Cordoba (Argentina). AB - Allele and genotype frequencies for nine STRs loci included in the AmpFlSTR Profiler Plus kit (D3S1385, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820), were determined from urban and countryside population of Cordoba (Argentina). All loci meet the Hardy-Weinberg expectation, and there is little evidence for alleles association between these nine loci. The results demonstrate that these loci can be useful for databasing purposes in human identification and parentage testing in the population of Cordoba (Argentina). PMID- 12590063 TI - Population genetic study of eight short tandem repeat loci CSF1PO, TPOX, TH01, F13A01, FESFPS, vWA, F13B and LPL in the Western Romanian population. AB - Allele frequencies for eight tetranucleotide short tandem repeat (STR) loci CSF1PO, TPOX, TH01, F13A01, FESFPS, vWA, F13B, LPL-were obtained from a population sample of 105-122 unrelated individuals born in Transylvania and Banat (Romania). PMID- 12590064 TI - Population data of Galicia (NW Spain) on the new Y-STRs DYS437, DYS438, DYS439, GATA A10, GATA A7.1, GATA A7.2, GATA C4 and GATA H4. AB - Haplotype, allele frequencies and population data of eight Y-chromosome STR loci, DYS437, DYS438, DYS439, GATA A10, GATA A7.1, GATA A7.2, GATA C4 and GATA H4, were determined from a sample of 212 unrelated male individuals from Galicia (NW of Spain). PMID- 12590065 TI - Comments on "an unusual case of sexual assault on an infant: an intraperitoneal candle in a 20-month-old-girl". PMID- 12590066 TI - Protection of the environment from ionising radiation. PMID- 12590067 TI - Protection of the environment from ionising radiation: ethical issues. AB - The paper identifies some of the main ethical issues concerning the protection of the environment from radiation and suggests ways in which ethics can aid in developing a system of protection. After a presentation of background on ethical theory and environmental ethics, three main issues related to environmental protection are discussed: First, the question of valuing the environment and implications for the definition of harm and monetary valuation of environmental goods; second, difficulties with scientific uncertainty and applications of the precautionary principle; and third, issues concerned with the distribution of risk and its relevance for participation in decision-making. In summary, the paper argues that there are strong ethical grounds to provide for the protection of the environment and that, all other things being equal, there is no reason to treat ionising radiation differently to other environmental stressors. PMID- 12590068 TI - Principles and issues in radiological ecological risk assessment. AB - This paper provides a bridge between the fields of ecological risk assessment (ERA) and radioecology by presenting key biota dose assessment issues identified in the US Department of Energy's Graded Approach for Evaluating Radiation Doses to Aquatic and Terrestrial Biota in a manner consistent with the US Environmental Protection Agency's framework for ERA. Current radiological ERA methods and data are intended for use in protecting natural populations of biota, rather than individual members of a population. Potentially susceptible receptors include vertebrates and terrestrial plants. One must ensure that all media, radionuclides (including short-lived radioactive decay products), types of radiations (i.e., alpha particles, electrons, and photons), and pathways (i.e., internal and external contamination) are combined in each exposure scenario. The relative biological effectiveness of alpha particles with respect to deterministic effects must also be considered. Expected safe levels of exposure are available for the protection of natural populations of aquatic biota (10 mGy d(-1)) and terrestrial plants (10 mGy d(-1)) and animals (1 mGy d(-1)) and are appropriate for use in all radiological ERA tiers, provided that appropriate exposure assumptions are used. Caution must be exercised (and a thorough justification provided) if more restrictive limits are selected, to ensure that the supporting data are of high quality, reproducible, and clearly relevant to the protection of natural populations. PMID- 12590069 TI - Derivation of a screening methodology for evaluating radiation dose to aquatic and terrestrial biota. AB - The United States Department of Energy (DOE) currently has in place a radiation dose standard for the protection of aquatic animals, and is considering additional dose standards for terrestrial biota. These standards are: 10 mGy/d for aquatic animals, 10 mGy/d for terrestrial plants, and, 1 mGy/d for terrestrial animals. Guidance on suitable approaches to the implementation of these standards is needed. A screening methodology, developed through DOE's Biota Dose Assessment Committee (BDAC), serves as the principal element of DOE's graded approach for evaluating radiation doses to aquatic and terrestrial biota. Limiting concentrations of radionuclides in water, soil, and sediment were derived for 23 radionuclides. Four organism types (aquatic animals; riparian animals; terrestrial animals; and terrestrial plants) were selected as the basis for development of the screening method. Internal doses for each organism type were calculated as the product of contaminant concentration, bioaccumulation factor(s) and dose conversion factors. External doses were calculated based on the assumption of immersion of the organism in soil, sediment, or water. The assumptions and default parameters used provide for conservative screening values. The screening methodology within DOE's graded approach should prove useful in demonstrating compliance with biota dose limits and for conducting screening assessments of radioecological impact. It provides a needed evaluation tool that can be employed within a framework for protection of the environment. PMID- 12590070 TI - A kinetic-allometric approach to predicting tissue radionuclide concentrations for biota. AB - Allometry, or the biology of scaling, is the study of size and its consequences. It has become a useful tool for comparative physiology. There are several allometric equations that relate body size to many parameters, including ingestion rate, lifespan, inhalation rate, home range and more. While these equations were originally derived from empirical observations, there is a growing body of evidence that these relationships have their origins in the dynamics of energy transport mechanisms. As part of an ongoing effort by the Department of Energy in developing generic methods for evaluating radiation dose to biota, we have examined the utility of applying allometric techniques to predicting radionuclide tissue concentration across a large range of terrestrial and riparian species of animals. This particular study examined 23 radionuclides. Initial investigations suggest that the allometric approach can provide a useful tool to derive limiting values of uptake and elimination factors for animals. PMID- 12590071 TI - A probabilistic approach to obtaining limiting estimates of radionuclide concentration in biota. AB - The US Department of Energy has developed a graded approach for evaluating radiation doses to biota. Limiting concentrations of radionuclides in water, soil, and sediment were derived for twenty-three radionuclides. Four organism types (aquatic animals, riparian animals, terrestrial animals, and terrestrial plants) were selected as the basis for method development. While environmental transfer data needed for deriving biota tissue concentrations are available for aquatic animals and terrestrial plants, less information is available for terrestrial and riparian organisms. Two methods were applied and examined for their ability to provide estimates of organism:soil or organism:water concentration factors in lieu of measured data. The kinetic/allometric approach combined with a parameter uncertainty analysis provides a needed method to estimate concentration factors across multiple species with limited input data. PMID- 12590072 TI - Generic performance assessment for a deep repository for low and intermediate level waste in the UK--a case study in assessing radiological impacts on the natural environment. AB - Concentrations of radionuclides in soil and surface water, taken from a generic performance assessment of a repository for low and intermediate level radioactive waste, assumed to be located in the UK, have been used as the basis for a case study in assessing radiological impacts on the natural environment. Simplified descriptions of the terrestrial and aquatic ecosystem types likely to be impacted have been developed. A scoping assessment has identified (226)Ra, (210)Po, (234)U, (230)Th and (238)U as having the highest potential for impact, with doses from internally incorporated alpha emitters as being potentially of particular importance. These nuclides, together with (36)Cl and (129)I (which have proved to be of importance in radiological risk assessments for humans) were included in a more detailed dose assessment. A basic methodology for dose assessment of ecosystems is described, and has been applied for the defined impacted ecosystems. Paucity of published data on concentration factors prevented a more detailed assessment for terrestrial ecosystems. For the aquatic ecosystem, a more detailed assessment was possible and highest calculated absorbed dose rates (weighted for the likely higher biological effectiveness of alpha radiation) were about 6.5 microGy h(-1). We conclude that harm to the impacted ecosystems is unlikely and make the observation that the lack of concentration factor or transfer factor data for a sufficiently wide range of species, ecosystems and nuclides appears to be the principal obstacle to establishing a comprehensive framework for the application of radiological protection to ecosystems. PMID- 12590073 TI - Environmental biodosimetry: a biologically relevant tool for ecological risk assessment and biomonitoring. AB - Biodosimetry, the estimation of received doses by determining the frequency of radiation-induced chromosome aberrations, is widely applied in humans acutely exposed as a result of accidents or for clinical purposes, but biodosimetric techniques have not been utilized in organisms chronically exposed to radionuclides in contaminated environments. The application of biodosimetry to environmental exposure scenarios could greatly improve the accuracy, and reduce the uncertainties, of ecological risk assessments and biomonitoring studies, because no assumptions are required regarding external exposure rates and the movement of organisms into and out of contaminated areas. Furthermore, unlike residue analyses of environmental media, environmental biodosimetry provides a genetically relevant biomarker of cumulative lifetime exposure. Symmetrical chromosome translocations can impact reproductive success, and could therefore prove to be ecologically relevant as well. We describe our experience in studying aberrations in the yellow-bellied slider turtle as an example of environmental biodosimetry. PMID- 12590074 TI - Radiological effects on populations of Oligochaeta in the Chernobyl contaminated zone. AB - A detailed investigation of 3 populations of Oligochaete species (Dero obtusa, Nais pseudobtusa and Nais pardalis) has been carried out in a contaminated lake of the close-in Chernobyl zone and in a control lake. Hydrochemical indices and concentrations of heavy metals, chloro-organic compounds and (90)Sr in bottom sediments have been measured. Absorbed doses were calculated on the basis of the results of radiochemical analysis and assessed directly with thermoluminescent dosimeters (TLD). Stimulation of paratomous division (asexual reproduction) was found in one species of worm (D. obtusa), and activation of sexual reproduction in the two other species studied. An increase in the amount of cytogenetic damage in the somatic cells of worms from the contaminated lake was found and an attempt was made to assess the relative contributions of radiation and chemical exposure on the basis of analyses of inter-cellular aberration distributions and the types of chromosome aberrations observed in the cells. PMID- 12590075 TI - Genetic consequences of radioactive contamination by the Chernobyl fallout to agricultural crops. AB - The genetic consequences of radioactive contamination by the fallout to agricultural crops after the accident at the Chernobyl NPP in 1986 have been studied. In the first, acute, period of this accident, when the absorbed dose was primarily due to external beta- and gamma-irradiation, the radiation injury of agricultural crops, according to the basic cytogenetic tests, resembled the effect produced by acute gamma-irradiation at comparable doses. The yield of cytogenetic damage in leaf meristem of plants grown in the 10-km zone of the ChNPP in 1987-1989 (the period of chronic, lower level radiation exposure) was shown to be enhanced and dependent on the level of radioactive contamination. The rate of decline with time in cytogenetic damage induced by chronic exposure lagged considerably behind that of the radiation exposure. Analysis of genetic variability in three sequential generations of rye and wheat revealed increased cytogenetic damage in plants exposed to chronic irradiation during the 2nd and 3rd years. PMID- 12590076 TI - Bioindication of the anthropogenic effects on micropopulations of Pinus sylvestris, L. in the vicinity of a plant for the storage and processing of radioactive waste and in the Chernobyl NPP zone. AB - Results of a comparative analysis of the frequency and spectrum of cytogenetic anomalies are presented for reproductive (seeds) and vegetative (needles) samples taken from Scotch pine (Pinus sylvestris, L.) micropopulations growing at sites with differing levels of radioactive contamination in the Chernobyl NPP 30 km zone, and at the location of a facility for the processing and storage of radioactive wastes (the 'Radon' LWPE, near the town of Sosnovy Bor in the Leningrad Region). The data obtained indicate the presence of genotoxic contaminants in the environment of the tree micropopulations. Chemical toxins make the main contribution to the environmental contamination in the Sosnovy Bor area as compared with the influence of ionising radiation in the Chernobyl 30 km zone. The higher radioresistance of seeds of Scotch pine growing on the area of the 'Radon' LWPE and in the centre of Sosnovy Bor town was revealed with acute gamma-radiation. PMID- 12590077 TI - A possible approach for the assessment of radiation effects on populations of wild organisms in radionuclide-contaminated environments? AB - It is frequently asserted that measures to protect the biotic environment from increased radiation exposures arising from human activities should be focussed on the population rather than the individual. It is, however, difficult (if not impossible) to identify any population-specific attributes that can be affected by radiation exposure directly rather than through the mediation of direct, known and identifiable effects in individual organisms. Indeed, it is often conceded that this difficulty forces attention to be refocussed onto the effects in individuals. Regulatory controls on radioactive waste management and disposal could then be implemented to ensure that any radiation effects in individual native plants and animals remain at, or below, some acceptable level (yet to be defined). Nevertheless, the question remains as to whether such controls would also provide for the protection of the population. An answer to this question depends on the availability of a model that allows the integration of the known effects of radiation exposure on the mortality, morbidity, fertility and fecundity of individuals into an assessment of the possible impact at the population level. The utility of one such approach, the Leslie Matrix Model, is explored in respect of a fish population (the plaice, Pleuronectes platessa). This initial implementation of the population model is simplistic (and, certainly, environmentally unrealistic), but it is concluded that the output from the model does provide some insights into how the population might respond to radiation-induced changes in individual attributes, and that further development in the direction of increased realism is fully warranted. PMID- 12590109 TI - Phytochemistry meets genome analysis, and beyond. PMID- 12590110 TI - Plant metabolomics: large-scale phytochemistry in the functional genomics era. AB - Metabolomics or the large-scale phytochemical analysis of plants is reviewed in relation to functional genomics and systems biology. A historical account of the introduction and evolution of metabolite profiling into today's modern comprehensive metabolomics approach is provided. Many of the technologies used in metabolomics, including optical spectroscopy, nuclear magnetic resonance, and mass spectrometry are surveyed. The critical role of bioinformatics and various methods of data visualization are summarized and the future role of metabolomics in plant science assessed. PMID- 12590111 TI - Metabolome diversity: too few genes, too many metabolites? AB - The multitude of metabolites found in living organisms and the calculated, unexpected small number of genes identified during genome sequencing projects discomfit biologists. Several processes on the transcription and translation level lead to the formation of isoenzymes and can therefore explain at least parts of this surprising result. However, poor enzyme specificity may also contribute to metabolome diversity. In former studies, when enzymes were isolated from natural sources, impure protein preparations were hold responsible for broad enzyme specificity. Nowadays, highly purified enzymes are available by molecular biological methods such as heterologous expression in host organisms and they can be thoroughly analyzed. During biochemical analysis of heterologously expressed enzymes poor specificity was observed for enzymes involved in fruit ripening, e.g. in flavour and color formation. Surprisingly broad specificity was shown for the reactants in the case of alcohol acyl-CoA transferase, O-methyltransferase, glucosyltransferase, P450 monooxygenases as well as polyketide synthases and for the product in the case of monoterpene synthases. Literature data confirm the assumption of limited specificity for enzymes involved in metabolism and bioformation of secondary metabolites. It is concluded that metabolome diversity is caused by low enzyme specificity but availability of suitable substrates due to compartmentation has also taken into account. PMID- 12590112 TI - Metabolomic analysis of the consequences of cadmium exposure in Silene cucubalus cell cultures via 1H NMR spectroscopy and chemometrics. AB - Several essential and non-essential metals (typically those from periods 4, 5 and 6 in groups 11-15 in the periodic table) are commonly detoxified in higher plants by complexation with phytochelatin. The genetic and gross metabolic basis of metal tolerance in plants is, however, poorly understood. Here, we have analyzed plant cell extracts using 1H NMR spectroscopy combined with multivariate statistical analysis of the data to investigate the biochemical consequences of Cd(2+) exposure in Silene cucubalus cell cultures. Principal components analysis of 1H NMR spectra showed clear discrimination between control and Cd(2+) dosed groups, demonstrating the metabolic effects of Cd(2+) and thus allowing the identification of increases in malic acid and acetate, and decreases in glutamine and branched chain amino acids as consequences of Cd(2+) exposure. This work shows the value of NMR-based metabolomic approaches to the determination of biochemical effects of pollutants in naturally selected populations. PMID- 12590113 TI - Chemometric discrimination of unfractionated plant extracts analyzed by electrospray mass spectrometry. AB - Metabolic fingerprints were obtained from unfractionated Pharbitis nil leaf sap samples by direct infusion into an electrospray ionization mass spectrometer. Analyses took less than 30 s per sample and yielded complex mass spectra. Various chemometric methods, including discriminant function analysis and the machine learning methods of artificial neural networks and genetic programming, could discriminate the metabolic fingerprints of plants subjected to different photoperiod treatments. This rapid automated analytical procedure could find use in a variety of phytochemical applications requiring high sample throughput. PMID- 12590114 TI - Lactofen induces isoflavone accumulation and glyceollin elicitation competency in soybean. AB - Lactofen, the active ingredient of the soybean disease resistance-inducing herbicide, Cobra, induces large accumulations of isoflavone conjugates and aglycones in soybean tissues. The predominant isoflavones induced in cotyledon tissues are daidzein (and its conjugates) and formononetin and glycitein aglycones. The latter two isoflavones are usually present only at very low levels in soybean seedling tissues. In leaves, the predominant lactofen-induced isoflavones are daidzein and formononetin aglycones and the malonyl-glucosyl conjugate of genistein. Isoflavone induction also occurs in cells distal to the point of treatment, but is only weakly systemic. Lactofen also induces elicitation competency, the capacity of soybean cells to accumulate the pterocarpan phytoalexin glyceollin in response to glucan elicitors from the cell wall of the pathogen Phytophthora sojae. Comparison of the activity of a series of diphenyl ether herbicides demonstrated that while all diphenyl ethers tested induced some degree of elicitation competency, only certain ones induced isoflavone accumulation in the absence of glucan elicitor. As a group the diphenyl ethers are thought to inhibit protoporhyrinogen oxidase, eventually leading to singlet oxygen generation. Another singlet oxygen generator, rose bengal, also induced elicitation competency, but little isoflavone accumulation. It is hypothesized that diphenyl ether-induced activated oxygen species mimic some aspects of hypersensitive cell death, which leads to elicitation competency in infected tissues. PMID- 12590115 TI - Metabolic networks of Cucurbita maxima phloem. AB - Metabolomic analysis aims at a comprehensive characterization of biological samples. Yet, biologically meaningful interpretations are often limited by the poor spatial and temporal resolution of the acquired data sets. One way to remedy this is to limit the complexity of the cell types being studied. Cucurbita maxima Duch. vascular exudates provide an excellent material for metabolomics in this regard. Using automated mass spectral deconvolution, over 400 components have been detected in these exudates, but only 90 of them were tentatively identified. Many amino compounds were found in vascular exudates from leaf petioles at concentrations several orders of magnitude higher than in tissue disks from the same leaves, whereas hexoses and sucrose were found in far lower amounts. In order to find the expected impact of assimilation rates on sugar levels, total phloem composition of eight leaves from four plants was followed over 4.5 days. Surprisingly, no diurnal rhythm was found for any of the phloem metabolites that was statistically valid for all eight leaves. Instead, each leaf had its own distinct vascular exudate profile similar to leaves from the same plant, but clearly different from leaves harvested from plants at the same developmental stage. Thirty to forty per cent of all metabolite levels of individual leaves were different from the average of all metabolite profiles. Using metabolic co regulation analysis, similarities and differences between the exudate profiles were more accurately characterized through network computation, specifically with respect to nitrogen metabolism. PMID- 12590116 TI - Construction and application of a mass spectral and retention time index database generated from plant GC/EI-TOF-MS metabolite profiles. AB - The non-supervised construction of a mass spectral and retention time index data base (MS/RI library) from a set of plant metabolic profiles covering major organs of potato (Solanum tuberosum), tobacco (Nicotiana tabaccum), and Arabidopsis thaliana, was demonstrated. Typically 300-500 mass spectral components with a signal to noise ratio > or =75 were obtained from GC/EI-time-of-flight (TOF)-MS metabolite profiles of methoxyaminated and trimethylsilylated extracts. Profiles from non-sample controls contained approximately 100 mass spectral components. A MS/RI library of 6205 mass spectral components was accumulated and applied to automated identification of the model compounds galactonic acid, a primary metabolite, and 3-caffeoylquinic acid, a secondary metabolite. Neither MS nor RI alone were sufficient for unequivocal identification of unknown mass spectral components. However library searches with single bait mass spectra of the respective reference substance allowed clear identification by mass spectral match and RI window. Moreover, the hit lists of mass spectral searches were demonstrated to comprise candidate components of highly similar chemical nature. The search for the model compound galactonic acid allowed identification of gluconic and gulonic acid among the top scoring mass spectral components. Equally successful was the exemplary search for 3-caffeoylquinic acid, which led to the identification of quinic acid and of the positional isomers, 4-caffeoylquinic acid, 5-caffeoylquinic acid among other still non-identified conjugates of caffeic and quinic acid. All identifications were verified by co-analysis of reference substances. Finally we applied hierarchical clustering to a complete set of pair-wise mass spectral comparisons of unknown components and reference substances with known chemical structure. We demonstrated that the resulting clustering tree depicted the chemical nature of the reference substances and that most of the nearest neighbours represented either identical components, as judged by co-elution, or conformational isomers exhibiting differential retention behaviour. Unknown components could be classified automatically by grouping with the respective branches and sub-branches of the clustering tree. PMID- 12590117 TI - Taxus metabolomics: methyl jasmonate preferentially induces production of taxoids oxygenated at C-13 in Taxus x media cell cultures. AB - Cells from suspension cultures of Taxus cuspidata were extracted with pentane as a source of relatively non-polar taxoids. Of the 13 taxoids identified in this fraction, eight were oxygenated at C-14 and two had not been previously described. These taxoids, along with existing taxoid standards, were employed to profile the metabolites of Taxus x media cv. Hicksii cell suspension cultures induced with methyl jasmonate to produce paclitaxel (Taxol). The majority of the taxoid metabolites produced in these induced cultures were oxygenated at C-13, and not C-14. PMID- 12590118 TI - The role of chemical fingerprinting: application to Ephedra. AB - Ephedra sinica, known as Ma Huang, is one of the oldest medicinal herbs in Traditional Chinese Medicine (TCM). Preparations, namely teas, of E. sinica have been used for over 5000 years as a stimulant and as an antiasthmatic. In the West, extracts of E. sinica, E. intermedia or E. equisetina are most commonly used in dietary supplements as a stimulant and to promote weight loss. More than 50 species of Ephedra are native to both hemispheres, but the detection of ephedrine alkaloids has been limited to species in Eurasia. Currently, methods exist to quantitate the ephedrine alkaloids in extracts of plant material or dietary supplements, but the methods are not able to verify the extract is of an Ephedra species. Reverse phase high performance liquid chromatography with photodiode array detection was applied for the chemical fingerprinting of the Ephedra species. Two regions of comparison were determined in the chromatograms at 320 nm. The series of peaks between 52 and 64 min confirms an Ephedra species is being analyzed. The aforementioned peaks also could distinguish between Ephedra species from Eurasia, North America and South America. Peaks at ca. 57 and 59 min were isolated and determined to be two new compounds, 4-(2 eicosyloxycarbonyl-vinyl)-benzoic acid and 4-(2-docosyloxycarbonyl-vinyl)-benzoic acid respectively. Authentication of ground plant material as Ephedra can be achieved by this chemical fingerprinting method. PMID- 12590119 TI - Metabolic fingerprinting of salt-stressed tomatoes. AB - The aim of this study was to adopt the approach of metabolic fingerprinting through the use of Fourier transform infrared (FT-IR) spectroscopy and chemometrics to study the effect of salinity on tomato fruit. Two varieties of tomato were studied, Edkawy and Simge F1. Salinity treatment significantly reduced the relative growth rate of Simge F1 but had no significant effect on that of Edkawy. In both tomato varieties salt-treatment significantly reduced mean fruit fresh weight and size class but had no significant affect on total fruit number. Marketable yield was however reduced in both varieties due to the occurrence of blossom end rot in response to salinity. Whole fruit flesh extracts from control and salt-grown tomatoes were analysed using FT-IR spectroscopy. Each sample spectrum contained 882 variables, absorbance values at different wavenumbers, making visual analysis difficult and therefore machine learning methods were applied. The unsupervised clustering method, principal component analysis (PCA) showed no discrimination between the control and salt-treated fruit for either variety. The supervised method, discriminant function analysis (DFA) was able to classify control and salt-treated fruit in both varieties. Genetic algorithms (GA) were applied to identify discriminatory regions within the FT-IR spectra important for fruit classification. The GA models were able to classify control and salt-treated fruit with a typical error, when classifying the whole data set, of 9% in Edkawy and 5% in Simge F1. Key regions were identified within the spectra corresponding to nitrile containing compounds and amino radicals. The application of GA enabled the identification of functional groups of potential importance in relation to the response of tomato to salinity. PMID- 12590120 TI - An optimized protocol for metabolome analysis in yeast using direct infusion electrospray mass spectrometry. AB - A method for the global analysis of yeast intracellular metabolites, based on electrospray mass spectrometry (ES-MS), has been developed. This has involved the optimization of methods for quenching metabolism in Saccharomyces cerevisiae and extracting the metabolites for analysis by positive-ion electrospray mass spectrometry. The influence of cultivation conditions, sampling, quenching and extraction conditions, concentration step, and storage have all been studied and adapted to allow direct infusion of samples into the mass spectrometer and the acquisition of metabolic profiles with simultaneous detection of more than 25 intracellular metabolites. The method, which can be applied to other micro organisms and biological systems, may be used for comparative analysis and screening of metabolite profiles of yeast strains and mutants under controlled conditions in order to elucidate gene function via metabolomics. Examples of the application of this analytical strategy to specific yeast strains and single-ORF yeast deletion mutants generated through the EUROFAN programme are presented. PMID- 12590121 TI - Identification and quantification of carotenoids, tocopherols and chlorophylls in commonly consumed fruits and vegetables. AB - The carotenoid, tocopherol and chlorophyll metabolic profiles and content of a selection of fruits and vegetables found commonly in the diet, have been determined using a rapid RP-HPLC technique with on-line PDA detection. Information gathered from the screening of secondary plant metabolites is vital for the accurate determination of the dietary intake of these micro-nutrients, and in the development of comprehensive food tables. Determination of basal levels is also necessary for the rational engineering of health-promoting phytochemicals in food crops. In addition this approach can also be applied to the routine screening of products to determine metabolic differences between varieties and cultivars, as well as between genetically modified and the corresponding non-genetically modified tissue. PMID- 12590122 TI - Assessment of 1H NMR spectroscopy and multivariate analysis as a technique for metabolite fingerprinting of Arabidopsis thaliana. AB - An approach to metabolite fingerprinting of crude plant extracts that utilizes 1H nuclear magnetic resonance (NMR) spectroscopy and multivariate statistics has been tested. Using ecotypes of Arabidopsis thaliana as experimental material, a method has been developed for the rapid analysis of unfractionated polar plant extracts, enabling the creation of reproducible metabolite fingerprints. These fingerprints could be readily stored and compared by a variety of chemometric methods. Comparison by principal component analysis using SIMCA-P allowed the generation of residual NMR spectra of the compounds that contributed significantly to the differences between samples. From these plots, conclusions were drawn with respect to the identity and relative levels of metabolites differing between samples. PMID- 12590123 TI - Monitoring changes in anthocyanin and steroid alkaloid glycoside content in lines of transgenic potato plants using liquid chromatography/mass spectrometry. AB - Transgenic potato plants overexpressing and repressing enzymes of flavonoids biosynthesis were created and analyzed. The selected plants clearly showed the expected changes in anthocyanins synthesis level. Overexpression of a DNA encoding dihydroflavonol 4-reductase (DFR) in sense orientation resulted in an increase in tuber anthocyanins, a 4-fold increase in petunidin and pelargonidin derivatives. A significant decrease in anthocyanin level was observed when the plant was transformed with a corresponding antisense construct. The transformation of potato plants was also accompanied by significant changes in steroid alkaloid glycosides (SAG) level in transgenic potato tuber. The changes in SAGs content was not dependent on flavonoid composition in transgenic potato. However, in an extreme situation where the highest (DFR11) or the lowest (DFRa3) anthocyanin level was detected the positive correlation with steroid alkaloid content was clearly visible. It is suggested that the changes in SAGs content resulted from chromatin stressed upon transformation. A liquid chromatography/mass spectrometry (LC/MS) system with electrospray ionization was applied for profiling qualitative and quantitative changes of steroid alkaloid glycosides in tubers of twelve lines of transgenic potato plants. Except alpha chaconine and alpha-solanine, in the extracts from dried tuber skin alpha solamargine and alpha-solasonine, triglycosides of solasonine, were identified in minor amounts, triglycosides of solanidine dehydrodimers were also recognized. PMID- 12590124 TI - Metabonomics classifies pathways affected by bioactive compounds. Artificial neural network classification of NMR spectra of plant extracts. AB - The biochemical mode-of-action (MOA) for herbicides and other bioactive compounds can be rapidly and simultaneously classified by automated pattern recognition of the metabonome that is embodied in the 1H NMR spectrum of a crude plant extract. The ca. 300 herbicides that are used in agriculture today affect less than 30 different biochemical pathways. In this report, 19 of the most interesting MOAs were automatically classified. Corn (Zea mays) plants were treated with various herbicides such as imazethapyr, glyphosate, sethoxydim, and diuron, which represent various biochemical modes-of-action such as inhibition of specific enzymes (acetohydroxy acid synthase [AHAS], protoporphyrin IX oxidase [PROTOX], 5 enolpyruvylshikimate-3-phosphate synthase [EPSPS], acetyl CoA carboxylase [ACC ase], etc.), or protein complexes (photosystems I and II), or major biological process such as oxidative phosphorylation, auxin transport, microtubule growth, and mitosis. Crude isolates from the treated plants were subjected to 1H NMR spectroscopy, and the spectra were classified by artificial neural network analysis to discriminate the herbicide modes-of-action. We demonstrate the use and refinement of the method, and present cross-validated assignments for the metabolite NMR profiles of over 400 plant isolates. The MOA screen also recognizes when a new mode-of-action is present, which is considered extremely important for the herbicide discovery process, and can be used to study deviations in the metabolism of compounds from a chemical synthesis program. The combination of NMR metabolite profiling and neural network classification is expected to be similarly relevant to other metabonomic profiling applications, such as in drug discovery. PMID- 12590125 TI - Metabolomics and differential gene expression in anthocyanin chemo-varietal forms of Perilla frutescens. AB - We have investigated metabolite profiles and gene expression in two chemo varietal forms, red and green forms, of Perilla frutescens var. crispa. Striking difference in anthocyanin content was observed between the red and green forms. Anthocyanin, mainly malonylshisonin, was highly accumulated in the leaves of the red form but not in the green form. Less obvious differences were also observed in the stems. However, there was no remarkable difference in the contents and patterns of flavones and primary metabolites such as inorganic anions, organic anions and amino acids. These results suggest that only the regulation of anthocyanin production, but not that of other metabolites, differs in red and green forms. Microscopic observation and immunohistochemical studies indicated that the epidermal cells of leaves and stems are the sites of accumulation of anthocyanins and localization of anthocyanidin synthase protein. By differential display of mRNA from the leaves of red and green forms, we could identify several genes encoding anthocyanin-biosynthetic enzymes and presumptive regulatory proteins. The possible regulatory network leading to differential anthocyanin accumulation in a form-specific manner is discussed. PMID- 12590126 TI - Regulation of floral scent production in petunia revealed by targeted metabolomics. AB - Petunia hybrida line W115 (Mitchell) has large white flowers that produce a pleasant fragrance. By applying solid phase micro extraction (SPME) techniques coupled to GC-MS analysis, volatile emission was monitored in vivo using a targeted metabolomics approach. Mature flowers released predominantly benzenoid compounds of which benzaldehyde, phenylacetaldehyde, methylbenzoate, phenylethylalcohol, iso-eugenol and benzylbenzoate were most abundant. This emission had a circadian rhythm reaching its maximum at dusk. During petal limb expansion two sesquiterpenes were emitted by the petunia flowers, tentatively identified as germacrene D and cadina-3,9-diene. In vitro analysis showed that the petal limbs and stigma were the main producers of the benzenoids and sesquiterpenes, respectively. Moreover, comparison of in vivo and in vitro analysis indicated that volatiles were not stored during periods of low emission but rather were synthesized de novo. DNA-microarray analysis revealed that genes of the pathways leading to the production of volatile benzenoids were upregulated late during the day, preceding the increase of volatile emission. RNA-gel blot analyses confirmed that the levels of phenylalanine ammonia lyase (PAL) and S adenosyl methionine (SAM) synthase transcripts increased towards the evening. Our results suggest that the circadian production of volatile benzenoids in petunia W115 is, at least partly, regulated at the transcript level. PMID- 12590127 TI - Factors affecting the robustness of metabolite fingerprinting using 1H NMR spectra. AB - 1H NMR spectroscopy is one of the techniques whose potential is currently being explored in the emerging field of metabolomics. It is a non-targeted method, producing signals for all proton-containing chemical species. For crude plant materials the spectra are always complex, with many signals overlapping. Hence a most suitable approach for analysing them is 'metabolite fingerprinting', which is aimed at highlighting compositional similarities and exploring the overall natural variability in a population of samples. The most commonly used method for this is principal component analysis (PCA), as it allows the whole spectral trace to be analysed and the vast quantity of information to be simplified. In this paper we investigate whether there are factors which may affect the NMR spectra in a way that subsequently decreases the robustness of the metabolite fingerprinting by PCA. Imperfections in the signal registration (i.e. inconsistency of the peak position) are generally detrimental to analysing whole traces by multivariate methods. The sources of such problems are illustrated through specially designed repeatability studies using potato and tomato samples, and the analysis of a tea dataset containing many samples. Careful sample preparation can help to limit peak shifts; for instance here by attempting to control the pH of the extracts. In addition, some compounds are susceptible to interactions affecting their chemical shifts and mathematical alignment of peaks may be necessary. Lastly factors such as resolution can also affect analyses and must be carefully adjusted. Our choice of examples aims to raise awareness of potential problems. We do not question the validity of the NMR approach, but point out those areas where special care may need to be taken. PMID- 12590129 TI - Uncensored exchange of scientific results. PMID- 12590130 TI - PNAS policy on publication of sensitive material in the life sciences. PMID- 12590131 TI - Proteoglycan UDP-galactose:beta-xylose beta 1,4-galactosyltransferase I is essential for viability in Drosophila melanogaster. AB - Heparan and chondroitin sulfates play essential roles in growth factor signaling during development and share a common linkage tetrasaccharide structure, GlcAbeta1,3Galbeta1,3Galbeta1,4Xylbeta1-O-Ser. In the present study, we identified the Drosophila proteoglycan UDP-galactose:beta-xylose beta1,4 galactosyltransferase I (dbeta4GalTI), and determined its substrate specificity. The enzyme transferred a Gal to the -beta-xylose (Xyl) residue, confirming it to be the Drosophila ortholog of human proteoglycan UDP-galactose:beta-xylose beta1,4-galactosyltransferase I. Then we established UAS-dbeta4GalTI-IR fly lines containing an inverted repeat of dbeta4GalTI ligated to the upstream activating sequence (UAS) promoter, a target of GAL4, and observed the F(1) generation of the cross between the UAS-dbeta4GalTI-IR fly and the Act5C-GAL4 fly. In the F(1), double-stranded RNA of dbeta4GalTI is expressed ubiquitously under the control of a cytoplasmic actin promoter to induce the silencing of the dbeta4GalTI gene. The expression of the target gene was disrupted specifically, and the degree of interference was correlated with phenotype. The lethality among the progeny proved that beta4GalTI is essential for viability. This study is the first to use reverse genetics, RNA interference, to study the Drosophila glycosyltransferase systematically. PMID- 12590132 TI - Alleviation of PC4-mediated transcriptional repression by the ERCC3 helicase activity of general transcription factor TFIIH. AB - Positive cofactor 4 (PC4), originally identified as a transcriptional coactivator, possesses the ability to suppress promoter-driven as well as nonspecific transcription via its DNA binding activity. Previous studies showed that the repressive activity of PC4 on promoter-driven transcription is alleviated by transcription factor TFIIH, possibly through one of its enzymatic activities. Using recombinant TFIIH, we have analyzed the role of TFIIH for alleviating PC4-mediated transcriptional repression and determined that the excision repair cross complementing (ERCC3) helicase activity of TFIIH is the enzymatic activity that alleviates PC4-mediated repression via beta-gamma bond hydrolysis of ATP. In addition, the alleviation does not require either ERCC2 helicase or cyclin-dependent kinase 7 kinase activity. We also show that, as complexed within TFIIH, the cyclin-dependent kinase 7 kinase does not possess the activity to phosphorylate PC4. Thus, TFIIH appears to protect promoters from PC4 mediated repression by relieving the topological constraint imposed by PC4 through the ERCC3 helicase activity rather than by reducing the repressive activity of PC4 via its phosphorylation. PMID- 12590133 TI - Conditional expression of SV40 T-antigen in mouse cardiomyocytes facilitates an inducible switch from proliferation to differentiation. AB - Studies of cardiac muscle gene expression and signaling have been hampered by the lack of immortalized cardiomyocyte cell lines capable of proliferation and irreversible withdrawal from the cell cycle. With the goal of creating such cell lines, we generated transgenic mice using cardiac-specific cis-regulatory elements from the mouse Nkx2.5 gene to drive the expression of a simian virus 40 large T-antigen (TAg) gene flanked by sites for recombination by Cre recombinase. These transgenic mice developed tumors within the ventricular myocardium. Cells isolated from these tumors expressed cardiac markers and proliferated rapidly during serial passage in culture, without apparent senescence. However, they were unable to exit the cell cycle and failed to exhibit morphological features of terminal differentiation. Introduction of Cre recombinase to these cardiac cell lines by adenoviral delivery resulted in the elimination of TAg expression, accompanied by rapid cessation of cell division, and increase in cell size without an apparent induction of cellular differentiation. Incubation of cells lacking TAg in serum-deficient media with various pharmacological agents (norepinephrine, phenylephrine, or bone morphogenetic protein-2/4) or constitutively active calcium/calmodulin-dependent protein kinase I and/or calcineurin led to the formation of sarcomeres and up-regulation of cardiac genes involved in excitation-contraction coupling. The combination of TAg expression under the control of an early cardiac promoter and Cre-mediated recombination allowed us to derive an immortal cell line from the ventricular myocardium that could be controllably withdrawn from the cell cycle. The conditional expression of TAg in this manner permits propagation and regulated growth termination of cell types that are otherwise unable to be maintained in cell culture and may have applications for cardiac repair technologies. PMID- 12590134 TI - Slp4-a/granuphilin-a inhibits dense-core vesicle exocytosis through interaction with the GDP-bound form of Rab27A in PC12 cells. AB - Slp4-a (synaptotagmin-like protein 4-a)/granuphilin-a is specifically localized on dense-core vesicles in PC12 cells and negatively controls dense-core vesicle exocytosis through specific interaction with Rab27A via the N-terminal Slp homology domain (SHD) (Fukuda, M., Kanno, E., Saegusa, C., Ogata, Y., and Kuroda, T. S. (2002) J. Biol. Chem. 277, 39673-39678). However, the mechanism of the inhibition by Slp4-a has never been elucidated at the molecular level and is still a matter of controversy. In this study, I discovered an unexpected biochemical property of Slp4-a, that Slp4-a, but not other Rab27 effectors reported thus far, is capable of interacting with both Rab27A(T23N), a dominant negative form that mimics the GDP-bound form, and Rab27A(Q78L), a dominant active form that mimics the GTP-bound form, whereas Slp4-a specifically recognizes the GTP-bound form of Rab3A and Rab8A and does not recognize their GDP-bound form. I show by deletion and mutation analyses that the TGDWFY sequence in SHD2 is essential for Rab27A(T23N) binding, whereas SHD1 is involved in Rab27A(Q78L) binding. I further show by immunoprecipitation and cotransfection assays that Munc18-1, but not syntaxin IA, directly interacts with the C-terminal domain of Slp4-a in a Rab27A-independent manner. Expression of Slp4-a mutants that lack Rab27A(T23N) binding activity (i.e. specific binding to Rab27A(Q78L)) completely reverses the inhibitory effect of the wild-type Slp4-a on high KCl-dependent neuropeptide Y secretion in PC12 cells. The results strongly indicate that interaction of Slp4-a with the GDP-bound form of Rab27A, not with syntaxin IA or Munc18-1, is the primary reason that Slp4-a expression inhibits dense core vesicle exocytosis in PC12 cells. PMID- 12590135 TI - The histone deacetylase 9 gene encodes multiple protein isoforms. AB - Histone deacetylases (HDACs) perform an important function in transcriptional regulation by modifying the core histones of the nucleosome. We have now fully characterized a new member of the Class II HDAC family, HDAC9. The enzyme contains a conserved deacetylase domain, represses reporter activity when recruited to a promoter, and utilizes histones H3 and H4 as substrates in vitro and in vivo. HDAC9 is expressed in a tissue-specific pattern that partially overlaps that of HDAC4. Within the human hematopoietic system, expression of HDAC9 is biased toward cells of monocytic and lymphoid lineages. The HDAC9 gene encodes multiple protein isoforms, some of which display distinct cellular localization patterns. For example, full-length HDAC9 is localized in the nucleus, but the isoform lacking the region encoded by exon 7 is in the cytoplasm. HDAC9 interacts and co-localizes in vivo with a number of transcriptional repressors and co-repressors, including TEL and N-CoR, whose functions have been implicated in the pathogenesis of hematological malignancies. These results suggest that HDAC9 plays a role in hematopoiesis; its deregulated expression may be associated with some human cancers. PMID- 12590136 TI - Identification of multiple RNA features that influence CCR4 deadenylation activity. AB - The CCR4 family proteins are 3'-5'-deadenylases that function in the first step of the degradation of poly(A) mRNA. Here we report the purification to homogeneity of the yeast CCR4 protein and the analysis of its substrate specificities. CCR4 deadenylated a 7N+23A substrate (seven nucleotides followed by 23 A residues) in a distributive manner. Only small differences in CCR4 activity for different A length substrates were observed until only 1 A residue remained. Correspondingly, the K(m) for a 25N+2A substrate was found to be at least 20-fold lower than that for a 26N+1A substrate, although their V(max) values differed by only 2-fold. In addition, the total length of the RNA was found to contribute to CCR4 activity: up to 17 nucleotides (not necessarily poly(A)) could be recognized by CCR4. Poly(U), poly(C), and poly(G) were also found to be 12-30-fold better inhibitors of CCR4 compared with poly(A), supporting the observation that CCR4 contains a non-poly(A)-specific binding site. Surprisingly, even longer substrates (>/=45 nucleotides) stimulated CCR4 to become a processive enzyme, suggesting that CCR4 undergoes an additional transition in the presence of such substrates. CCR4 also displayed no difference in its activity with capped or uncapped RNA substrates. These results indicate that CCR4 recognition of its RNA substrates involves several features of the RNA that could be sites in vivo for controlling the rate of specific mRNA deadenylation. PMID- 12590137 TI - Bone-related genes expressed in advanced malignancies induce invasion and metastasis in a genetically defined human cancer model. AB - We employed a genetically defined human cancer model to investigate the contributions of two genes up-regulated in several cancers to phenotypic changes associated with late stages of tumorigenesis. Specifically, tumor cells expressing two structurally unrelated bone-related genes, osteonectin and osteoactivin, acquired a highly invasive phenotype when implanted intracranially in immunocompromised mice. Mimicking a subset of gliomas, tumor cells invaded brain along blood vessels and developed altered vasculature at the brain-tumor interface, suggesting that production of those two proteins by tumor cells may create a complex relationship between invading tumor and vasculature co-opted during tumor invasion. Interestingly, the same tumor cells formed massive spontaneous metastases when implanted subcutaneously. This dramatic alteration in tumor phenotype indicates that cellular microenvironment plays an important role in defining the specific effects of those gene products in tumor behavior. In vitro examination of tumor cells expressing either osteonectin or osteoactivin revealed that there was no impact on cellular growth or death but increased invasiveness and expression of MMP-9 and MMP-3. Specific pharmacologic inhibitors of MMP-2/9 and MMP-3 blocked the increased in vitro invasion associated with osteoactivin expression, but only MMP-3 inhibition altered the invasive in vitro phenotype mediated by osteonectin. Results from this genetically defined model system are supported by similar findings obtained from several established tumor cell lines derived originally from human patients. In sum, these results reveal that the expression of a single bone-related gene can dramatically alter or modify tumor cell behavior and may confer differential growth characteristics in different microenvironments. Genetically defined human cancer models offer useful tools in functional genomics to define the roles of specific genes in late stages of carcinogenesis. PMID- 12590138 TI - Distinct protein kinase C isoforms mediate regulation of vascular endothelial growth factor expression by A2A adenosine receptor activation and phorbol esters in pheochromocytoma PC12 cells. AB - Vascular endothelial growth factor (VEGF) stimulates angiogenesis during development and in disease. In pheochromocytoma (PC12) cells, VEGF expression is regulated by A(2A) adenosine receptor (A(2A)AR) activation. The present work examines the underlying signaling pathway. The adenylyl cyclase-protein kinase A cascade has no role in the down-regulation of VEGF mRNA induced by the A(2A)AR agonist, 2-[4-[(2-carboxyethyl)phenyl]ethylamino]-5'-N-ethylcarboxamidoadenosine (CGS21680). Conversely, 6-h exposure of cells to either phorbol 12-myristate 13 acetate (PMA) or protein kinase C (PKC) inhibitors mimicked the CGS21680-induced down-regulation. PMA activated PKCalpha, PKCepsilon, and PKCzeta, and CGS21680 activated PKCepsilon and PKCzeta as assessed by cellular translocation. By 6 h, PMA but not CGS21680 decreased PKCalpha and PKCepsilon expression. Neither compound affected PKCzeta levels. Following prolonged PMA treatment to down regulate susceptible PKC isoforms, CGS21680 but not PMA inhibited the cobalt chloride induction of VEGF mRNA. The proteasome inhibitor, MG-132, abolished PMA- but not CGS21680-induced down-regulation of VEGF mRNA. Phorbol 12,13-diacetate reduced VEGF mRNA levels while down-regulating PKCepsilon but not PKCalpha expression. In cells expressing a dominant negative PKCzeta construct, CGS21680 was unable to reduce VEGF mRNA. Together, the findings suggest that phorbol ester induced down-regulation of VEGF mRNA occurs as a result of a reduction of PKCepsilon activity, whereas that mediated by the A(2A)AR occurs following deactivation of PKCzeta. PMID- 12590139 TI - Novel docosatrienes and 17S-resolvins generated from docosahexaenoic acid in murine brain, human blood, and glial cells. Autacoids in anti-inflammation. AB - Docosahexaenoic acid (DHA, C22:6) is highly enriched in brain, synapses, and retina and is a major omega-3 fatty acid. Deficiencies in this essential fatty acid are reportedly associated with neuronal function, cancer, and inflammation. Here, using new lipidomic analyses employing high performance liquid chromatography coupled with a photodiode-array detector and a tandem mass spectrometer, a novel series of endogenous mediators was identified in blood, leukocytes, brain, and glial cells as 17S-hydroxy-containing docosanoids denoted as docosatrienes (the main bioactive member of the series was 10,17S docosatriene) and 17S series resolvins. These novel mediators were biosynthesized via epoxide-containing intermediates and proved potent (pico- to nanomolar range) regulators of both leukocytes reducing infiltration in vivo and glial cells blocking their cytokine production. These results indicate that DHA is the precursor to potent protective mediators generated via enzymatic oxygenations to novel docosatrienes and 17S series resolvins that each regulate events of interest in inflammation and resolution. PMID- 12590140 TI - Transcriptional activation of the human inducible nitric-oxide synthase promoter by Kruppel-like factor 6. AB - Nitric oxide is a ubiquitous free radical that plays a key role in a broad spectrum of signaling pathways in physiological and pathophysiological processes. We have explored the transcriptional regulation of inducible nitric-oxide synthase (iNOS) by Kruppel-like factor 6 (KLF6), an Sp1-like zinc finger transcription factor. Study of serial deletion constructs of the iNOS promoter revealed that the proximal 0.63-kb region can support a 3-6-fold reporter activity similar to that of the full-length 16-kb promoter. Within the 0.63-kb region, we identified two CACCC sites (-164 to -168 and -261 to -265) that bound KLF6 in both electrophoretic mobility shift and chromatin immunoprecipitation assays. Mutation of both these sites abrogated the KLF6-induced enhancement of the 0.63-kb iNOS promoter activity. The binding of KLF6 to the iNOS promoter was significantly increased in Jurkat cells, primary T lymphocytes, and COS-7 cells subjected to NaCN-induced hypoxia, heat shock, serum starvation, and phorbol 12 myristate 13-acetate/ ionophore stimulation. Furthermore, in KLF6-transfected and NaCN-treated COS-7 cells, there was a 3-4-fold increase in the expression of the endogenous iNOS mRNA and protein that correlated with increased production of nitric oxide. These findings indicate that KLF6 is a potential transactivator of the human iNOS promoter in diverse pathophysiological conditions. PMID- 12590141 TI - In vitro synthesis of lactose permease to probe the mechanism of membrane insertion and folding. AB - Insertion and folding of polytopic membrane proteins is an important unsolved biological problem. To study this issue, lactose permease, a membrane transport protein from Escherichia coli, is transcribed, translated, and inserted into inside-out membrane vesicles in vitro. The protein is in a native conformation as judged by sensitivity to protease, binding of a monoclonal antibody directed against a conformational epitope, and importantly, by functional assays. By exploiting this system it is possible to express the N-terminal six helices of the permease (N(6)) and probe changes in conformation during insertion into the membrane. Specifically, when N(6) remains attached to the ribosome it is readily extracted from the membrane with urea, whereas after release from the ribosome or translation of additional helices, those polypeptides are not urea extractable. Furthermore, the accessibility of an engineered Factor Xa site to Xa protease is reduced significantly when N(6) is released from the ribosome or more helices are translated. Finally, spontaneous disulfide formation between Cys residues at positions 126 (Helix IV) and 144 (Helix V) is observed when N(6) is released from the ribosome and inserted into the membrane. Moreover, in contrast to full-length permease, N(6) is degraded by FtsH protease in vivo, and N(6) with a single Cys residue at position 148 does not react with N-ethylmaleimide. Taken together, the findings indicate that N(6) remains in a hydrophilic environment until it is released from the ribosome or additional helices are translated and continues to fold into a quasi-native conformation after insertion into the bilayer. Furthermore, there is synergism between N(6) and the C-terminal half of permease during assembly, as opposed to assembly of the two halves as independent domains. PMID- 12590142 TI - An amino acid cluster around the essential Glu-14 is part of the substrate- and proton-binding domain of EmrE, a multidrug transporter from Escherichia coli. AB - EmrE is a small multidrug transporter (110 amino acids long) from Escherichia coli that extrudes various drugs in exchange with protons, thereby rendering bacteria resistant to these compounds. Glu-14 is the only charged membrane embedded residue in EmrE and is evolutionarily highly conserved. This residue has an unusually high pK and is an essential part of the binding domain, shared by substrates and protons. The occupancy of the binding domain is mutually exclusive, and, as such, this provides the molecular basis for the coupling between substrate and proton fluxes. Systematic cysteine-scanning mutagenesis of the residues in the transmembrane segment (TM1), where Glu-14 is located, reveals an amino acid cluster on the same face of TM1 as Glu-14 that is part of the substrate- and proton-binding domain. Substitutions at most of these positions yielded either inactive mutants or mutants with modified affinity to substrates. Substitutions at the Ala-10 position, one helix turn away from Glu-14, yielded mutants with modified affinity to protons and thereby impaired in the coupling of substrate and proton fluxes. Taken as a whole, the results strongly support the concept of a common binding site for substrate and protons and stress the importance of one face of TM1 in substrate recognition, binding, and H(+)-coupled transport. PMID- 12590143 TI - Diversity of oligopeptide transport specificity in Lactococcus lactis species. A tool to unravel the role of OppA in uptake specificity. AB - The specific oligopeptide transport system Opp is essential for growth of Lactococcus lactis in milk. We examined the biodiversity of oligopeptide transport specificity in the L. lactis species. Six strains were tested for (i) consumption of peptides during growth in a chemically defined medium and (ii) their ability to transport these peptides. Each strain demonstrated some specific preferences for peptide utilization, which matched the specificity of peptide transport. Sequencing of the binding protein OppA in some strains revealed minor differences at the amino acid level. The differences in specificity were used as a tool to unravel the role of the binding protein in transport specificity. The genes encoding OppA in four strains were cloned and expressed in L. lactis MG1363 deleted for its oppA gene. The substrate specificity of these engineered strains was found to be similar to that of the L. lactis MG1363 parental strain, whichever oppA gene was expressed. In situ binding experiments demonstrated the ability of OppA to interact with non-transported peptides. Taken together, these results provide evidence for a new concept. Despite that fact that OppA is essential for peptide transport, it is not the (main) determinant of peptide transport specificity in L. lactis. PMID- 12590144 TI - Solution structure and function of the "tandem inactivation domain" of the neuronal A-type potassium channel Kv1.4. AB - Cumulative inactivation of voltage-gated (Kv) K(+) channels shapes the presynaptic action potential and determines timing and strength of synaptic transmission. Kv1.4 channels exhibit rapid "ball-and-chain"-type inactivation gating. Different from all other Kvalpha subunits, Kv1.4 harbors two inactivation domains at its N terminus. Here we report the solution structure and function of this "tandem inactivation domain" using NMR spectroscopy and patch clamp recordings. Inactivation domain 1 (ID1, residues 1-38) consists of a flexible N terminus anchored at a 5-turn helix, whereas ID2 (residues 40-50) is a 2.5-turn helix made up of small hydrophobic amino acids. Functional analysis suggests that only ID1 may work as a pore-occluding ball domain, whereas ID2 most likely acts as a "docking domain" that attaches ID1 to the cytoplasmic face of the channel. Deletion of ID2 slows inactivation considerably and largely impairs cumulative inactivation. Together, the concerted action of ID1 and ID2 may promote rapid inactivation of Kv1.4 that is crucial for the channel function in short term plasticity. PMID- 12590145 TI - AF-6 controls integrin-mediated cell adhesion by regulating Rap1 activation through the specific recruitment of Rap1GTP and SPA-1. AB - In the present study, we showed that SPA-1, a Rap1 GTPase-activating protein (GAP), was bound to a cytoskeleton-anchoring protein AF-6. SPA-1 and AF-6 were co immunoprecipitated in the 293T cells transfected with both cDNAs as well as in normal thymocytes. In vitro binding studies using truncated fragments and their mutants suggested that SPA-1 was bound to the PDZ domain of AF-6 via probable internal PDZ ligand motif within the GAP-related domain. The motif was conserved among Rap1 GAPs, and it was shown that rapGAP I was bound to AF-6 comparably with SPA-1. RapV12 was also bound to AF-6 via the N-terminal domain, and SPA-1 and RapV12 were co-immunoprecipitated only in the presence of AF-6, indicating that they could be brought into close proximity via AF-6 in cells. Immunostaining analysis revealed that SPA-1 and RapV12 were co-localized with AF-6 at the cell attachment sites. In HeLa cells expressing SPA-1 in a tetracycline-regulatory manner, expression of AF-6 inhibited endogenous Rap1GTP and beta(1) integrin mediated cell adhesion to fibronectin in SPA-1-induced conditions, whereas it affected neither of them in SPA-1-repressed conditions. These results suggested that AF-6 could control integrin-mediated cell adhesion by regulating Rap1 activation through the recruitment of both SPA-1 and Rap1GTP via distinct domains. PMID- 12590146 TI - Cloning and characterization of a novel Na+-dependent glucose transporter (NaGLT1) in rat kidney. AB - To identify novel transporters in the kidney, we have constructed an mRNA data base composed of 1000 overall clones by random sequencing of a male rat kidney cDNA library. After a BLAST search, approximately 40% of the clones were unknown and/or unannotated and were screened by measuring the uptake of various compounds using Xenopus oocytes. One clone stimulated the uptake of alpha-methyl-d glucopyranoside and therefore was termed rat Na(+)-dependent glucose transporter 1 (rNaGLT1). The rNaGLT1 cDNA (2173 bp) has an open reading frame encoding a 484 amino acid protein, showing <22% homology to known SGLT and GLUT glucose transporters. alpha-Methyl-d-glucopyranoside uptake by rNaGLT1 cRNA-injected oocytes showed saturability, with an apparent K(m) of 3.7 mm and a coupling ratio of 1:1 with Na(+). rNaGLT1 mRNA was expressed predominantly in the kidney upon Northern blot analysis and reverse transcription-PCR. Reverse transcription-PCR in microdissected nephron segments revealed that rNaGLT1 mRNA was primarily localized in the proximal tubules. A clear signal corresponding to rNaGLT1 protein was recognized in the brush-border (but not basolateral) membrane fraction by immunoblot analysis. The rNaGLT1 mRNA level in the kidney was significantly higher than rat SGLT1 and SGLT2 mRNA levels. These findings suggest that rNaGLT1 is a novel Na(+)-dependent glucose transporter with low substrate affinity that mediates tubular reabsorption of glucose. PMID- 12590147 TI - Role of the connecting peptide in insulin biosynthesis. AB - In single-chain insulins (SCIs), the C terminus of the insulin B-chain is contiguous with the N terminus of the A-chain, connected by a short bioengineered linker sequence. SCIs have been proposed to offer potential benefit for gene therapy of diabetes (Lee, H. C., Kim, S. J., Kim, K. S., Shin, H. C., and Yoon, J. W. (2000) Nature 408, 483-488) yet relatively little is known about their folding, intracellular transport, or secretion from mammalian cells. Because SCIs can be considered as mutant proinsulin (with selective shortening of the 35-amino acid connecting peptide that normally includes two sets of flanking dibasic residues), they offer insights into understanding the role of the connecting peptide in insulin biosynthesis. Herein we have explored the relationship of the linker sequence to SCI biosynthesis, folding, and intracellular transport in transiently transfected HEK293 or Chinese hamster ovary cells or in stably transfected AtT20 cells. Despite previous reports that direct linkage of B- and A chains produces a structure isomorphous with authentic two-chain insulin, we find that constructs with short linkers tend to be synthesized at lower levels, with a significant fraction of molecules exhibiting improper disulfide bonding. Nevertheless, disulfide-mispaired isoforms from a number of different SCI constructs are secreted. While this suggests that a novel folded state goes unrecognized by secretory pathway quality control, we find that misfolded SCIs are detected at higher levels in Chinese hamster ovary cells with artificially activated unfolded protein response mediated by inducible overexpression of active ATF-6. Such a maneuver allows analysis of more seriously misfolded mutants with further foreshortening of the linker sequence or loss (by mutation) of the insulin interchain disulfide bonds. PMID- 12590148 TI - Flow cytometric bivariate analysis of DNA and cytokeratin in colorectal cancer. AB - Different opinions about flow cytometric estimates of DNA aneuploidy and/or S phase fraction (SPF) as supplementary prognostic markers in colorectal cancer are to some degree associated with methodology. Using univariate DNA analysis, we have previously investigated the DNA ploidy in colorectal cancer, its heterogeneity within and between tumors and its relation to survival. To improve detection of DNA aneuploid subpopulations and particularly estimation of their SPF's we investigated a method for bivariate DNA/cytokeratin analysis on fine needle aspirates of 728 frozen biopsies from 157 colorectal tumors. Unfixed aspirates were stained with propidium iodide and FITC-conjugated anti-cytokeratin antibody in a saponin-buffer. A significant association between SPF and debris was observed. There were no substantial difference in DNA ploidy patterns between univariate and bivariate measurements (concordance was 92-95%). No new DNA aneuploid subpopulations were detected in cytokeratin-gated compared to ungated or univariate histograms. Debris-adjusted SPF's of cytokeratin-gated histograms were significantly higher than of ungated histograms, also for subpopulations with DI>1.4 (p<0.0001). There was no significant association between SPF and survival. PMID- 12590149 TI - Microdissection and molecular analysis of single cells or small cell clusters in pathology and diagnosis--significance and challenges. PMID- 12590150 TI - Evaluation of prognostic factors following flow-cytometric DNA analysis after cytokeratin labelling: I. Breast cancer. AB - In gynecologic oncology valid prognostic factors are necessary to estimate the course of disease and to define biologically similar subgroups for analysis of therapeutic efficacy. The presented study is a prospective study concerning prognostic significance of DNA ploidy and S-phase fraction in breast cancer following enrichment of tumor cells by cytokeratin labelling. Epithelial cells were labeled by FITC-conjugated cytokeratin antibody (CK 5, 6, 8, and CK 17) prior to flow cytometric cell cycle analysis in 327 fresh specimens of primary breast cancer. Univariate analysis in breast cancer detected the prognostic significance of DNA-ploidy, S-phase fraction and CV (coefficient of variation) of G(0)G(1)-peak of tumor cells for clinical outcome, especially for nodal-negative patients. Multivariate analysis could not confirm prognostic evidence of DNA ploidy and S-phase fraction. In conclusion, in breast cancer no clinical significance for determination of DNA-parameters was found. PMID- 12590152 TI - Dependence of DNA-histograms on the sampling techniques in fine needle aspirates of the breast. AB - 48 fine needle aspiration biopsy (FNAB) samples from 25 breast cancer cases, originally used for cytodiagnosis were subjected to DNA cytometry. There were air dried smears stained with the MGG method, and samples stained with HE or PAP stain after 50% ethanol fixation and cytocentrifugation. Different sampling strategies were applied. Four methods were tested: method 1: cell groups measured, method 2: all cells measured, method 3: free cells measured, and method 4: atypical free cells measured. Method 4 showed most often DNA aneuploid histogram patterns, sampling method 1 had the highest number of DNA diploid histogram patterns. Diagnostic approaches may benefit from a sampling method detecting the hiding aneuploid cell population. Grading of neoplasm could potentially benefit from other approaches. PMID- 12590151 TI - Evaluation of prognostic factors following flow-cytometric DNA analysis after cytokeratin labelling: II. Cervical and endometrial cancer. AB - In gynecologic oncology valid prognostic factors are necessary to define biologically similar subgroups for analysis of therapeutic efficacy. This study is the first published prospective study concerning prognostic significance of DNA ploidy and S-phase fraction in cervical and endometrial cancer following enrichment of tumor cells by cytokeratin labelling. Epithelial cells were labeled by FITC-conjugated cytokeratin antibody (CK 5, 6, 8, and CK 17) prior to flow cytometric cell cycle analysis in 91 specimens of cervical cancer and 73 samples of endometrial cancer. In cervical cancer neither DNA-ploidy nor S-phase fraction were relevant prognostic parameters. But CV of the G(0)G(1)-peak showed prognostic relevance in cervical cancer cells, even in multivariate analysis. This interesting observation, however, seems to have no therapeutic consequence due to the small discrimination capacity of CV. In endometrial carcinoma, gross DNA-aneuploidy (DNA-index > 1.3) and a high percentage of proliferating cells (>75th percentile) were univariate and multivariate highly significant prognostic factors for recurrence-free survival. Especially DNA-aneuploidy (DI>1.3) is one of the most important independent molecular biological prognostic factors. While diagnostic curettage we could identify risk patients even preoperatively by determination of the prognostic factors like histologic tumor type, grading, cervical involvement and DNA-ploidy. Thereby these patients could be treated primarily in an oncologic center. In conclusion, our investigations showed that the determination of DNA-ploidy should be done in endometrial carcinoma. In cervical cancer no clinical significance for determination of DNA-parameters was found. PMID- 12590153 TI - Chromosomal regions in prostatic carcinomas studied by comparative genomic hybridization, hierarchical cluster analysis and self-organizing feature maps. AB - Comparative genomic hybridization (CGH) is an established genetic method which enables a genome-wide survey of chromosomal imbalances. For each chromosome region, one obtains the information whether there is a loss or gain of genetic material, or whether there is no change at that place. Therefore, large amounts of data quickly accumulate which must be put into a logical order. Cluster analysis can be used to assign individual cases (samples) to different clusters of cases, which are similar and where each cluster may be related to a different tumour biology. Another approach consists in a clustering of chromosomal regions by rewriting the original data matrix, where the cases are written as rows and the chromosomal regions as columns, in a transposed form. In this paper we applied hierarchical cluster analysis as well as two implementations of self organizing feature maps as classical and neuronal tools for cluster analysis of CGH data from prostatic carcinomas to such transposed data sets. Self-organizing maps are artificial neural networks with the capability to form clusters on the basis of an unsupervised learning rule. We studied a group of 48 cases of incidental carcinomas, a tumour category which has not been evaluated by CGH before. In addition we studied a group of 50 cases of pT2N0-tumours and a group of 20 pT3N0-carcinomas. The results show in all case groups three clusters of chromosomal regions, which are (i) normal or minimally affected by losses and gains, (ii) regions with many losses and few gains and (iii) regions with many gains and few losses. Moreover, for the pT2N0- and pT3N0-groups, it could be shown that the regions 6q, 8p and 13q lay all on the same cluster (associated with losses), and that the regions 9q and 20q belonged to the same cluster (associated with gains). For the incidental cancers such clear correlations could not be demonstrated. PMID- 12590154 TI - A monitoring/auditing mechanism for SSL/TLS secured service sessions in Health Care Applications. AB - This paper analyzes the SSL/TLS procedures and defines the functionality of a monitoring/auditing entity running in parallel with the protocol, which is decoding, checking the certificate and permitting session establishment based on the decoded certificate information, the network addresses of the endpoints and a predefined access list. Finally, this paper discusses how such a facility can be used for detection impersonation attempts in Health Care applications and provides case studies to show the effectiveness and applicability of the proposed method. PMID- 12590155 TI - Cross-sectional changes in an asymmetric tube with bearing on non-invasive pressure measurements. AB - This study aims at investigating the radial dimensional changes, as a result of an applied intraluminal pressure for an elastic tube with non-uniform cross section. The study is related to a method for non-invasive pressure measurement using the extracorporeal tube as part of the sensor. The intended application is for monitoring of pressure in the blood and dialysate tubes during haemodialysis. The intention is to find a tube cross-section geometry that results in an expansion of the tube so that it is suitable to use as a component in a pressure sensor. The tube should have high radial compliance and expand in a well-defined manner to be able to transfer the intraluminal pressure to a transducer element sensing the radial force. Radial expansion was studied experimentally for tubes with different cross-section geometries. For small tube expansions the resolution in the experimental measurements was not sufficient to study the radial expansion. In this case, numerical simulation was performed. We conclude that a tube with essentially elliptic outer surface and circular inner surface, with a relation of 1:2 between the size of the thin and thick wall, results in a radial expansion upon application of pressure indicating that this tube is suitable for use as part of a sensor. PMID- 12590156 TI - A flow visualization study of an anatomic coronary artery anastomosis model with an implant. AB - Flow Streamlining Devices is a new tool in Coronary Artery Bypass Grafting (CABG). They aim in: a) Performing a sutureless anastomosis to reduce thrombosis at the veno-arterial junction, and b) Providing a hemodynamically efficient scaffolding to reduce secondary flow disturbances. Thrombosis and flow disturbances are factors that have been reported as contributing factors to the development of intimal hyperplasia (IH) and failure of the graft. By reducing thrombosis and flow disturbances, it is expected that IH will be inhibited and the lifetime of the graft extended. To evaluate the hemodynamic benefits of such an implant, two models were designed and fabricated. One simulated the geometry of the conventional anastomosis without an implant, and the other simulated an anastomosis with a flow streamlining implant. Identical flow conditions relevant to a coronary anastomosis were imposed on both models and flow visualization was performed with dye injection and a digital camera. Results showed reduction of disturbances in the presence of the implant. This reduction seems to be favorable to hemodynamic streamlining which may create conditions that may inhibit the initialization of IH. However, the compliance and geometric mismatch between the anastomosis and the implant created a disturbance at the rigid compliant wall interface, which should be eliminated prior to clinical applications. PMID- 12590157 TI - Secure environment for real-time tele-collaboration on virtual simulation of radiation treatment planning. AB - A secure framework is described for real-time tele-collaboration on Virtual Simulation procedure of Radiation Treatment Planning. An integrated approach is followed clustering the security issues faced by the system into organizational issues, security issues over the LAN and security issues over the LAN-to-LAN connection. The design and the implementation of the security services are performed according to the identified security requirements, along with the need for real time communication between the collaborating health care professionals. A detailed description of the implementation is given, presenting a solution, which can directly be tailored to other tele-collaboration services in the field of health care. The pilot study of the proposed security components proves the feasibility of the secure environment, and the consistency with the high performance demands of the application. PMID- 12590158 TI - Experimental verification of conditions for near infrared spectroscopy (NIRS). AB - OBJECTIVES: in vitro assessment of the reproducibility and the optimal separation and position of the optodes in continuous wave (CW-) NIRS measurement of local inhomogeneities in absorption and/or scattering. METHODS: a CW- NIRS system (OXYMON) was used with laser diodes at wavelengths of 767 nm, 845 nm, 905 nm, 945 nm and 975 nm. For practical considerations (dimensions of neonatal head) the measurements were performed on a cylindrical tissue-equivalent phantom (70 mm diameter of base material with mua = 0.01 mm(-1) (800 nm) and mu's = 1.00 mm(-1) (800 nm)), containing rods with 10 x absorption, or 10 x scattering, and 5 x both Monte Carlo simulations were carried out of a cylinder with transport scattering coefficient mu's = 0.525 mm(-1) and absorption coefficient mua = 0.075 mm(-1) and two optode positions. RESULTS: reproducibility of repeated measurements (n = 10) was +/- 0.005 OD. Maximum OD in case of absorbing rod, and of absorbing + scattering rod was measured with optodes separated by 90 degrees and rod position angle symmetrically (45 degrees ) in between. Minimum OD for these rods was obtained with optodes at 150 degrees angle and rod position at 240 degrees (i.e. relative to transmitting optode position at 0 degrees ). A second maximum OD was obtained at an optode angle 180 degrees and rod position at 180 degrees. Maximum OD (i.e. attenuation) for the scattering rod was at optode separation angle of 90 degrees and rod at 0 degrees. Minimum OD for this case was obtained with optode angle of 180 degrees and rod positions around 80 degrees and 280 degrees. Maximum OD changes by absorbing rod were in the order of +0.12 OD and -0.04 OD, respectively. Simulations at an optode separation angle of 90 degrees showed a spatial sensitivity path enclosing the rod position at maximum absorption found experimentally. CONCLUSIONS: when considering the phantom as a realistic geometrical model for the neonatal head, it can be concluded that the optode position at 90 degrees angle would be optimal for detecting an inhomogeneity at 15 mm depth, i.e. the location of the periventricular white matter. Since the rods are relatively strongly different from the base material the question remains to be answered whether local ischemia, which might lead to irreversible brain damage, can be detected by CW-NIRS PMID- 12590159 TI - Enriching healthcare applications with cryptographic mechanisms and XML- based security services. AB - The paper presents the enrichment of an existing e-referral / e-prescription application within a Regional Healthcare Information Network with security functionality, solving current authentication, integrity, non-repudiation and confidentiality issues and thus significantly enhancing the overall system security, operability, applicability and user acceptance. The application makes use of an underlying PKI framework, in order to provide strong authentication, digital signature, encryption and time-stamping services. XML is used for the representation of the healthcare data itself, the encrypted form of this data, as well as the relevant data security information, following W3C standards. PMID- 12590160 TI - Apolipoprotein E influences amyloid-beta clearance from the murine periphery. AB - The relationship between amyloid-beta protein (Abeta) metabolism and Alzheimer's disease is currently poorly understood. While it is well known that the generation of Abeta results from enzymatic cleavage of its parent molecule, the amyloid beta protein precursor (AbetaPP), there is little information available regarding its in vivo clearance. The E4 isoform of apolipoprotein E (apoE) has been associated with poor clearance of Abeta under in vitro conditions. This is thought to be due to its poor ability to bind Abeta compared with the other common isoforms, apoE2 and apoE3. Although cell culture studies support the notion that Abeta clearance depends upon apoE isoform, validation of these findings requires Abeta clearance studies in vivo. In this study, we examined the clearance of Abeta in vivo from the periphery in mice that expressed apoE (C57BL/6J) or lacked apoE (APOE knockout). We measured the clearance of peripherally injected Abeta over time and additionally, the quantities sequestered by peripheral organs. Western blot analysis of the murine plasma indicated that the half-life of Abeta in the periphery was approximately 15 minutes. The livers of the C57BL/6J mice were found to have sequestered approximately 40% of the total injected Abeta at 90 minutes post-injection, whilst their kidneys contained 5% of the total injected Abeta. In contrast, the livers and kidneys of the APOE knockout animals were found to contain no detectable Abeta. These findings indicate that Abeta is rapidly removed from the plasma by murine peripheral tissues and the rate of its clearance is affected by apoE. PMID- 12590161 TI - Vitamin E deficiency does not induce compensatory antioxidant increases in central nervous system tissue of apolipoprotein E-deficient mice. AB - Compensatory upregulation in endogenous antioxidants has been shown to accompany certain genetic and dietary deficiencies that promote oxidative stress, including that related to Alzheimer's disease. We compared antioxidant levels in brain tissue of normal and transgenic mice lacking apolipoprotein E following dietary deprivation of vitamin E or folate. As described previously, ApoE-deficient mice displayed increased levels of the endogenous antioxidant glutathione as compared to normal mice, and increased these levels further following folate deprivation. By contrast, glutathione was depleted following vitamin E deprivation in brain tissue of normal and ApoE-deficient mice. TBAR analyses confirmed increased oxidative damage following vitamin E deprivation. However, combined deprivation of folate and vitamin E resulted in levels of glutathione intermediate between those observed following deprivation of either agent, indicating that the lack of compensatory increase in glutathione following vitamin E deprivation was not due to overt neurotoxicity. Similar results were observed for total antioxidant levels in brain tissue. The differential response to vitamin E and folate deprivation is consistent with the possibility that specific differences in oxidative damage may result from deficiencies in either of these agents. The lack of a compensatory response to vitamin E deprivation highlights the importance of dietary vitamin E in prevention of chronic neurodegeneration. PMID- 12590162 TI - Aggresome formation by mutant prion proteins: the unfolding role of proteasomes in familial prion disorders. AB - Although familial prion disorders are a direct consequence of mutations in the prion protein gene, the underlying mechanisms leading to neurodegeneration remain unclear. Potential pathogenic mechanisms include abnormal cellular metabolism of the mutant prion protein (PrP(M)), or destabilization of PrP(M) structure inducing a change in its conformation to the pathogenic PrP-scrapie (PrP(Sc)) form. To further clarify these mechanisms, we investigated the biogenesis of mutant PrP V203I and E211Q associated with Creutzfeldt-Jakob disease, and PrP Q212P associated with Gerstmann-Straussler-Scheinker syndrome in neuroblastoma cells. We report that all three PrP(M) forms accumulate similarly in the cytosol in response to proteasomal inhibition, and finally assemble as classical aggresomes. Since the three PrP(M) forms tested in this report are distinct, we propose that sequestration of misfolded PrP(M) into aggresomes is likely a general response of the cellular quality control that is not specific to a particular mutation in PrP. Moreover, since PrP has the remarkable ability to refold into PrP(Sc) that can subsequently replicate, PrP(M) sequestered in aggresomes may cause neurotoxicity by both direct and indirect pathways; directly through PrP(Sc) aggregates, and indirectly by depleting normal PrP, through induction of a cellular stress response, or by other undefined pathways. On the other hand, sequestered PrP(M) may be relatively inert, and cellular toxicity may be mediated by early intermediates in aggresome formation. Taken together, these observations demonstrate the role of proteasomes in the pathogenesis of familial prion disorders, and argue for further explanation of its mechanistic details. PMID- 12590163 TI - Advanced glycation endproducts cause lipid peroxidation in the human neuronal cell line SH-SY5Y. AB - Advanced glycation endproducts (AGEs), sugar-derived protein modifications and lipid peroxidation products are prominent features of Alzheimer's disease. AGEs accumulate on beta-amyloid plaques during the course of the disease and can exert chronic oxidative stress via receptor-mediated mechanisms. Lipid peroxidation products such as hydroxynonenal, further markers of oxidative stress, are also increased in Alzheimer's diesease. In this study we present evidence for a direct biochemical link between AGEs and lipid peroxidation. Our results show that AGEs induce lipid peroxidation in a neuronal cell line in a dose-dependant manner, and that blocking the specific AGE-receptor RAGE, as well as using different antioxidants (alpha-lipoic acid, N-acetylcysteine, 17 beta-estradiol or aminoguanidine) can reduce the AGE-mediated formation of lipid peroxidation products. Thus, both RAGE antagonists and scavengers of oxygen free radicals could be useful in protecting brain tissue from lipid peroxidation and its pathophysilogical consequences that occur in Alzheimer's disease. PMID- 12590164 TI - Aluminum and copper interact in the promotion of oxidative but not inflammatory events: implications for Alzheimer's disease. AB - The etiology of Alzheimer's Disease (AD) is multifactorial. It has been suggested that transition metals such as copper (Cu) and iron (Fe) as well as aluminum (Al) may be involved in the pathogenesis of the disorder. While Cu and Fe are redox active, Al only exists in the trivalent form and is redox-inert. We previously demonstrated that Al exposure causes an increase in inflammatory parameters in human glioblastoma T98G cells. In the present study we further demonstrate that co-exposure with Cu exacerbates the oxidative but not inflammatory effects of Al in this cell line. While Cu-induced reactive oxygen species (ROS) production was greatly enhanced in the presence of Al, TNF-alpha secretion induced by either metal was not further potentiated by simultaneous exposure to Al and Cu. Furthermore, exposure to both metals reduced the individual Al and Cu-induced activation of the immune-related transcription factor NF-kappaB. Therefore, while synergistic interaction between the two metals increases oxidative events, this does not lead to potentiation of Al-induced inflammation. Thus the ability of aluminum to promote inflammatory processes does not depend on an increase ROS production induced by interaction with transition metals. PMID- 12590165 TI - Cholinergic plasticity in hippocampus of individuals with mild cognitive impairment: correlation with Alzheimer's neuropathology. AB - Several recent studies indicate that activity of cholinergic enzymes in the cortex of people with mild cognitive impairment (MCI) and early Alzheimer's disease (AD) are preserved. We correlated levels of hippocampal choline acetyltransferase (ChAT) activity with the extent of AD lesions in subjects from the Religious Order Study, including cases with no cognitive impairment (NCI), MCI, and with mild to moderate AD. Hippocampal ChAT activity levels were also determined in a group of end-stage AD patients who were enrolled in the University of Pittsburgh Alzheimer's Disease Research Center. MCI subjects were characterized with increased hippocampal ChAT activity. This elevation was no longer present in mild AD cases, which were not different from NCI subjects. Severe AD cases showed markedly depleted hippocampal ChAT levels. In NCI, MCI, and mild-moderate AD, there was a positive correlation between hippocampal ChAT activity levels and progression of neuritic plaque pathology in entorhinal cortex and hippocampus. A significant elevation of hippocampal ChAT in the MCI group was found selectively in the limbic (i.e., entorhinal-hippocampal, III/IV) Braak stages. We hypothesize that cholinergic changes in the hippocampus of MCI subjects reflect a compensatory response to the progressive denervation of the hippocampus by lost entorhinal cortex input. Moreover, the present findings suggest that the short-term memory loss observed in MCI is not caused by cholinergic deficits; it more likely relates to disrupted entorhinal-hippocampal connectivity. PMID- 12590166 TI - Superoxide dismutase activity in the mongolian gerbil brain after acute poisoning with aluminum. AB - In this study, the activity of total superoxide dismutase was investigated in brains of adult Mongolian gerbils (Meriones unguiculatus) treated with aluminum. AlCl3x6H20, was given "per os" in the amount of 3.7 g/kg body weight. Animals were killed 24, 48, 72 and 96 hours after the treatment and SOD activity was measured in crude mitochondrial fractions of cortex, thalamus and hippocampus. The SOD activity was significantly elevated in all investigated brain regions 24 hours after aluminum administration. The most prominent increases (up to 200% of values in control animals) were detected in thalamus and hippocampus, whereas the activity was 165% of control value in the cortex. The SOD activity returned to control values in all regions investigated forty-eight hours after poisoning. A slight secondary increases in SOD activity were observed at 72 hours, reaching 171%, 148%, and 166% of control values in the thalamus, hippocampus and cortex, respectively, 96 hours after AlCl3x6H20 administration. We conclude that Al administration causes a biphasic stimulation of SOD activity in various brain regions over 96 hours, providing evidence that oxidative stress is involved in aluminum toxicity to the brain. PMID- 12590167 TI - Length of stay in skilled nursing facilities is longer for patients with dementia. AB - Studies estimate prolonged stays in acute and sub-acute facilities for patients with dementia. Actuarial projections suggest prolonged stays in long term care facilities for patients with dementia. To test these predictions, we assessed whether patients with dementia stay in skilled nursing facilities (SNF) longer than patients without dementia. We obtained medical records of 5,373 residents discharged from a SNF between 1996 and 2001. Residents were identified as having dementia by ICD-9 codes. Age, sex and length of stay (LOS), measured in days from admission to discharge or death, were gathered. Mean LOS for patients with dementia (92.7 +/- 313.0, n = 758) was significantly longer than non-demented patients (29.7 +/- 136.8, n = 4615, p < 0.001). In a subset of individuals who stayed until death, the mean LOS for patients with dementia (202.9 +/- 528.6, n = 195) also was significantly longer than for non-demented patients (91.8 +/- 300.5, n = 610, p < 0.001). LOS was increased for demented patients even within age groups. Thus, patients with dementia stay in SNFs significantly longer from entry until discharge or death. It is likely that demented patients enter for non physical, cognitive related reasons. These results may help families and institutions plan for long-term care. PMID- 12590168 TI - Nanotechnology: Emerging Developments and Early Detection of Cancer. A two-day workshop sponsored by the National Cancer Institute and the National Institute of Standards and Technology, August 30-31 2001, on the National Institute of Standards and Technology Campus, Gaithersburg, MD, USA. AB - A recent meeting jointly sponsored by the National Cancer Institute (NCI) and National Institute of Standards and Technology (NIST) brought together researchers active in nanotechnology and cancer molecular biology to discuss and evaluate the interface between disciplines. Emerging areas where nanotechnologies may impact cancer prevention and early cancer detection were elaborated by key researchers who catalyzed interdisciplinary dialogue aimed at fostering cross discipline communications and future collaboration. PMID- 12590169 TI - Calcified nanostructured silicon wafer surfaces for biosensing: effects of surface modification on bioactivity. AB - The growth of known biologically-relevant mineral phases on semiconducting surfaces is one strategy to explicitly induce bioactivity in such materials, either for sensing or drug delivery applications. In this work, we describe the use of a spark ablation process to fabricate deliberate patterns of Ca(10)(PO4)6(OH)2 on crystalline Si (calcified nanoporous silicon). These patterns have been principally characterized by scanning electron microscopy in conjunction with elemental characterization by energy dispersive x-ray analysis. This is followed by a detailed comparison of the effects of fibroblast adhesion and proliferation onto calcified nanoporous Si, calcified nanoporous Si derivatized with alendronate, as well as control samples of an identical surface area containing porous SiO2. Fibroblast adhesion and proliferation assays demonstrate that a higher density of cells grow on the Ca3(PO4)2/porous Si/SiO2 structures relative to the alendronate-modified surfaces and porous Si/SiO2 samples. PMID- 12590170 TI - Bioassays based on molecular nanomechanics. AB - Recent experiments have shown that when specific biomolecular interactions are confined to one surface of a microcantilever beam, changes in intermolecular nanomechanical forces provide sufficient differential torque to bend the cantilever beam. This has been used to detect single base pair mismatches during DNA hybridization, as well as prostate specific antigen (PSA) at concentrations and conditions that are clinically relevant for prostate cancer diagnosis. Since cantilever motion originates from free energy change induced by specific biomolecular binding, this technique is now offering a common platform for label free quantitative analysis of protein-protein binding, DNA hybridization DNA protein interactions, and in general receptor-ligand interactions. Current work is focused on developing "universal microarrays" of microcantilever beams for high-throughput multiplexed bioassays. PMID- 12590171 TI - Mechanical spectral signatures of malignant disease? A small-sample, comparative study of continuum vs. nano-biomechanical data analyses. AB - Thin sections from human breast biopsies were employed to perform a differential analysis of the ultrasound spectral responses from invasive ductal carcinoma and normal tissue. A non-destructive testing methodology was employed, yielding the reflection coefficients as function of frequency in the clinical ultrasound range. The spectral responses were simulated both in the context of continuum and nano-biomechanics, with the objective of quantifying the physical properties that determine the differences in the spectral signature of normal vs. malignant tissue. The properties that were employed for the theoretical reconstruction of the spectra were: the density, the continuum and the nanomechanical elastic constants, and the nanomechanical theory internodal distance. The latter is a measure of the depth-of-penetration of mechanical actions between contiguous tissue elements. Together with vectorial descriptors of the tissue spatial arrangement, the internodal distance variable affords the quantitative incorporation of tissue architectural data in the theoretical model. In this paper, the validity of the nanomechanical approach to tissue characterization is discussed, and its potential extensions to biomolecular marker-based cancer diagnostics and therapeutics are considered. PMID- 12590172 TI - Nanometer-scale pores: potential applications for analyte detection and DNA characterization. AB - Several classes of transmembrane protein ion channels function in vivo as sensitive and selective detection elements for analytes. Recent studies on single channels reconstituted into planar lipid bilayer membranes suggest that nanometer scale pores can be used to detect, quantitate and characterize a wide range of analytes that includes small ions and single stranded DNA. We briefly review here these studies and identify leaps in technology that, if realized, might lead to innovations for the early detection of cancer. PMID- 12590174 TI - Assessment of normal variability in peripheral blood gene expression. AB - Peripheral blood is representative of many systemic processes and is an ideal sample for expression profiling of diseases that have no known or accessible lesion. Peripheral blood is a complex mixture of cell types and some differences in peripheral blood gene expression may reflect the timing of sample collection rather than an underlying disease process. For this reason, it is important to assess study design factors that may cause variability in gene expression not related to what is being analyzed. Variation in the gene expression of circulating peripheral blood mononuclear cells (PBMCs) from three healthy volunteers sampled three times one day each week for one month was examined for 1,176 genes printed on filter arrays. Less than 1% of the genes showed any variation in expression that was related to the time of collection, and none of the changes were noted in more than one individual. These results suggest that observed variation was due to experimental variability. PMID- 12590173 TI - Utility of the blood for gene expression profiling and biomarker discovery in chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) is a debilitating illness lacking consistent anatomic lesions and eluding conventional laboratory diagnosis. Demonstration of the utility of the blood for gene expression profiling and biomarker discovery would have implications into the pathophysiology of CFS. The objective of this study was to determine if gene expression profiles of peripheral blood mononuclear cells (PMBCs) could distinguish between subjects with CFS and healthy controls. Total RNA from PBMCs of five CFS cases and seventeen controls was labeled and hybridized to 1764 genes on filter arrays. Gene intensity values were analyzed by various classification algorithms and nonparametric statistical methods. The classification algorithms grouped the majority of the CFS cases together, and distinguished them from the healthy controls. Eight genes were differentially expressed in both an age-matched case-control analysis and when comparing all CFS cases to all controls. Several of the differentially expressed genes are associated with immunologic functions (e.g., CMRF35 antigen, IL-8, HD protein) and implicate immune dysfunction in the pathophysiology of CFS. These results successfully demonstrate the utility of the blood for gene expression profiling to distinguish subjects with CFS from healthy controls and for identifying genes that could serve as CFS biomarkers. PMID- 12590175 TI - A feature set for cytometry on digitized microscopic images. AB - Feature extraction is a crucial step in most cytometry studies. In this paper a systematic approach to feature extraction is presented. The feature sets that have been developed and used for quantitative cytology at the Laboratory for Biomedical Image Analysis of the GSF as well as at the Center for Image Analysis in Uppsala over the last 25 years are described and illustrated. The feature sets described are divided into morphometric, densitometric, textural and structural features. The latter group is used to describe the eu- and hetero-chromatin in a way complementing the textural methods. The main goal of the paper is to bring attention to the need of a common and well defined description of features used in cyto- and histometrical studies. The application of the sets of features is shown in an overview of projects from different fields. Finally some rules of thumb for the design of studies in this field are proposed. Colour figures can be viewed on http://www.esacp.org/acp/2003/25-1/rodenacker.htm. PMID- 12590176 TI - Automated detection of working area of peripheral blood smears using mathematical morphology. AB - The paper presents a technique to automatically detect the working area of peripheral blood smears stained with May-Grunwuald Giemsa. The optimal area is defined as the well spread part of the smear. This zone starts when the erythrocytes stop overlapping (on the body film side) and finishes when the erythrocytes start losing their clear central zone (on the feather edge side). The approach yields a quick detection of this area in images scanned under low magnifying power (immersion objective x 25 or x 16). The algorithm consists of two stages. First, an image analysis procedure using mathematical morphology is applied for extracting the erythrocytes, the centers of erythrocytes and the erythrocytes with center. Second, the number of connected components from the three kinds of particles is counted and the coefficient of spreading rho(s) and the coefficient of overlapping rho(o) are calculated. The data from fourteen smears illustrate how the technique is used and its performance. Colour figures can be viewed on http://www.esacp.org/acp/2003/25-1/angulo.htm. PMID- 12590177 TI - Noncoding RNA transcripts. AB - Recent analyses of the human genome and available data about the other higher eukaryotic genomes have revealed that, in contrast to Eubacteria and Archaea, only a small fraction of the genetic material (ca 1.5%) codes for proteins. Most of genomic DNA and its RNA transcripts are involved in regulation of gene expression, which can be exerted at either the transcriptional level, controlling whether a gene is transcribed and to what extent, or at the post-translational level, regulating the fate of the transcribed RNA molecules, including their stability, efficiency of their translation and subcellular localization. Noncoding RNA genes produce functional RNA molecules (ncRNAs) rather than encoding proteins. These stable RNAs act by multiple mechanisms such as RNA-RNA base pairing, RNA-protein interactions and intrinsic RNA activity, as well as regulate diverse cellular functions, including RNA processing, mRNA stability, translation, protein stability and secretion. Non-protein-coding RNAs are known to play significant roles. Along with transfer RNAs, ribosomal RNAs and mRNAs, ncRNAs contribute to gene splicing, nucleotide modification, protein transport and regulation of gene expression. PMID- 12590178 TI - Saturating rye genetic map with amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) markers. AB - A linkage map of rye, previously developed using DS2 x RXL10 F2 mapping population, was enriched with 179 AFLP and 19 RAPD marker loci. The current map covers 1386 cM and contains 480 markers including 200 RFLPs, 179 AFLPs, 88 RAPDs, 12 protein loci and one dwarfing gene. AFLPs generated by EcoRI/MseI primer combinations were distributed over the entire genome as distinct loci or clusters of 2-14 tightly linked DNA fragments. New marker loci mapped distally to the existing framework, significantly increased coverage of chromosomes 1R, 2R and 5R. The average marker distance is now 2.9 cM, but in seven regions the closest markers are still more than 20 cM apart. A detailed description of the newly mapped AFLP and RAPD loci is presented. The relationship with other published rye maps is discussed. PMID- 12590179 TI - Isozymes in Aegilops kotschyi and Ae. biuncialis x Secale cereale hybrids and Ae. kotschyi x S. cereale amphiploids in relation to their parents. AB - Seven enzymatic systems in F1 Aegilops kotschyi and Ae. biuncialis x Secale cereale hybrids, Aegilops kotschyi x S. cereale amphiploids and their parental species (Ae. kotschyi, Ae. biuncialis and S. cereale) were analysed by starch and polyacrylamide gel electrophoresis. Five of them (phosphoglucose isomerase, glutamic oxalacetic transaminase, esterase, acid phosphatase, and diaphorase) were polymorphic and two (malic dehydrogenase and superoxide dismutase) were monomorphic. Several isophorms of phosphoclucose isomerase, esterase, acid phosphatase, and diaphorase were detected in some hybrids and amphiploids, but absent in the parents. The role of regulators, translocations and recombination is discussed in relation to the origin of these new isophorms. Some parental isozymes were absent both in hybrids and amphiploids, probably as a result of the suppression of structural genes in new combinations of the three genomes. PMID- 12590180 TI - Cytological investigations of the interspecific hybrids of Nicotiana tabacum L. x N. glauca Grah. AB - Interspecific amphihaploid and amphidiploid hybrids between Nicotiana glauca Grah. (2n = 24) and N. tabacum L. (2n = 48) cultivars BY 103 and K 326 were analysed. F1 amphihaploids (2n = 36) were viable and completely self- and cross sterile, and mostly univalents were present during meiosis (with pairing range from 0 to 5). In some meiocytes, meiotic irregularities were observed, such as sporadic chromatin bridges and formation of restitution nuclei. The resultant F1 hybrids were easily converted to amphidiploids (2n = 72) via colchicine treatment of seedlings. The number of univalents and the frequency of PMCs containing unpaired chromosomes indicated that amphidiploids N. tabacum cv. BY 103 or K 326 x N. glauca represented quite a high pairing category. However, they were male sterile because pollen mother cells were arrested at the tetrad stage. The termination of development of PMCs, and consequently male sterility, are very rare in this kind of tobacco hybrids. PMID- 12590181 TI - Variability of cathodic peroxidases in sugar beet (Beta vulgaris L.) cultivars. AB - Eighteen varieties of sugar beet (Beta vulgaris L.), originating from various European countries, were compared in respect of peroxidase variability level. They were cultivated in the same experimental plot. The cultivars differed in ploidy level: one variety was tetraploid, three were diploid and 14 varieties were triploid. The cathodic peroxidase system is controlled by four independent genes, of which only one is polymorphic. Consequently, the varieties were characterised by frequencies of 3 allozymes belonging to one locus. Only one variety proved to be fully monomorphic. Genetic similarities between the cultivars were illustrated by a dendrogram (UPGMA) and show different groups of varieties not related to their ploidy level. PMID- 12590182 TI - Estimation of the dominance merit in noninbred populations without recourse to its inverted relationship matrix. AB - This study presents a new approach to obtain dominance estimates without using the full Henderson's mixed model equations (MMEs) related to an additive plus dominance animal model. This reduction could decrease substantially the computing time and hence its cost. In contrast to a procedure that we proposed before, the method developed in this paper does not require D(-1) and provides best linear unbiased prediction (BLUP) of genetic values that is close to that given by processing the full MMEs. In the previous study, we also elaborated an algorithm (denoted xi-REML) in order to approximate restricted maximum likelihood estimation of variance components via the expectation maximization (EM) algorithm. The xi-REML algorithm has been modified to be adapted to our new resolution approach. Through a numerical example, we show that there is a good agreement between REML-(EM), xi-REML and modified xi-REML estimates and that the latter algorithm is more efficient than our first proposition in terms of computing time and memory conservation. PMID- 12590183 TI - Application of dual colour FISH for localisation of canine microsatellite markers in the Chinese raccoon dog (Nyctereutes procyonoides procyonoides) genome. AB - The present study shows new chromosomal localisation of four canine cosmid clones in the Chinese raccoon dog genome by using dual colour FISH. This approach facilitates rapid physical localisation of markers and will improve the determination of their order on chromosomes. The present new assignments increase the number of physically mapped markers in the Chinese raccoon dog to 25. PMID- 12590184 TI - Cystic fibrosis--a probable cause of Frederic Chopin's suffering and death. AB - Frederic Chopin--a great Polish composer and pianist--suffered from a chronic disease. Both during his life and after death physicians disagreed on the subject of Chopin's diagnosis. His contemporaries accepted the diagnosis of a disease common in the 18th century--tuberculosis. Description of new clinical entities provoked new dilemmas in the 20th century. In our opinion the most tenable seems to be the diagnosis of cystic fibrosis. In this work we present F. Chopin's case history and discuss cons and pros for cystic fibrosis as the cause of F. Chopin's suffering and death. PMID- 12590185 TI - Increased risk of the abdominal aortic aneurysm in carriers of the MTHFR 677T allele. AB - Abdominal aortic aneurysm (AAA) presents itself as a progressive dilation of the abdominal aorta, leading--if untreated--to rupture. It is a common disease of the elderly, with a complex etiology. Several genetic, biochemical and environmental factors are recognized as relevant for the pathogenesis of AAA. We determined the polymorphism of the MTHFR (methylenetetrahydrofolate reductase) gene within the fourth exon (C677T) in 63 patients with AAA and compared it to that in 75 subjects of the population sample. The frequencies of the C/C, C/T and T/T genotypes were 65%, 27%, and 8% in the population sample and 33%, 60%, and 6% in the patients. This corresponds to a 4.4-fold greater risk of AAA in subjects who have the 677C/T variant of MTHFR, as compared with those who are 677C/C (p < 0.0001; 95% CI=2.11-9.34). The frequency of allele MTHFR 677T in patients (0.37) was higher than in the population sample (0.21; p < 0.007). This association between the common allele of the MTHFR gene--MTHFR 677T--and the development of AAA suggests that elevated homocysteine (Hcy) may disturb the function of the aortic wall. The disturbance may involve enhancement of elastin degradation, the process enhanced by mild hyperhomocysteinemia in minipigs. The magnitude of this effect, which refers to the AAA patients unselected for familial occurrence, indicates that the disturbance of aortic wall physiology caused by the presence of the MTHFR 677T allele is greater than the effect of the earlier described allele disequilibrium at the polymorphic alleles of the PAI1 (plasminogen activator inhibitor 1) gene seen only in familial cases of AAA. PMID- 12590186 TI - COL1A1 mutation analysis in Lithuanian patients with osteogenesis imperfecta. AB - Osteogenesis imperfecta (OI) is a generalised disorder of connective tissue characterised by an increased fragility of bones and also manifested in other tissues containing collagen type I, by blue sclera, hearing loss, dentinogenesis imperfecta, hyperextensible joints, hernias and easy bruising. OI is dominantly inherited and results in >90% OI cases, caused by mutations in one of the two genes COL1A1 or COL1A2 coding for type I procollagen. The Lithuanian OI database comprises 147 case records covering the period of 1980 - 2001. Clinical and genealogical analysis of OI cases/families from Lithuania available for examination revealed 18 familial cases of OI type I and 22 sporadic cases: OI type II (3 cases), OI type III (11 cases) and OI type I (8 cases). As a result of their molecular genetic investigation, 11 mutations were identified in the COL1A1 gene in 13 unrelated patients. Of them, nine mutations (E500X, G481A, c.2046insCTCTCTAG, c.1668delT, c.1667insC, c.4337insC, IVS19+1G > A, IVS20-2A > G, IVS22-1G > T) appeared to be novel, i.e. not yet registered in the Human Type I and Type III Collagen Mutations Database (http://www.le.ac.uk/genetics/collagen). PMID- 12590187 TI - Three novel mutations in exon 21 encoding beta-cardiac myosin heavy chain. AB - Familial hypertrophic cardiomyopathy has a complex multigenic background. Previous work allowed to determine one of the gene loci responsible for this disease on chromosome 14 band q11-q12, and linked it to the alpha and beta cardiac myosin heavy chains. In this study we demonstrate changes in exon 21, coding for beta-myosin heavy chain. We described 4 patients from different families with an unequivocal diagnosis of hypertrophic cardiomyopathy based on the clinical picture. Direct sequencing of exon 21 revealed the presence of 5 novel mutations. Two of the mutations in codons 771 and 781 revealed in our study did not result in any changes in amino acid sequence. The next three were as follows: in codon 782 (AGC > GAC) transition responsible for Ser-->Asp substitution; in codon 779 (GAG > TAG) mutation that results in replacement of Glu-->Stop; in codon 774 (GAG > GTG) which is expressed as substitution of Glu- >Val. These mutations are located close to mutations identified and described in the literature, so they are likely to cause similar symptoms. PMID- 12590188 TI - Polymorphisms of the 5,10-methylenetetrahydrofolate and the methionine synthase reductase genes as independent risk factors for spina bifida. AB - We analyzed the role of the C677T polymorphism of the 5,10 methylenetetrahydrofolate and the A66G polymorphism of the methionine synthase reductase genes as risk factors for occurrence of spina bifida. The studied population included 106 mothers and 104 children from affected families, and a control group of 100 adults. We found statistically significant differences between the occurrence of the homozygosity in these polymorphisms in the groups of mothers and children with thoracolumbal defects (C677T polymorphism) and lumbosacral defects (A66G polymorphism). We postulate that these polymorphisms should be regarded as independent risk factors for spina bifida. PMID- 12590189 TI - The challenge of neuroscience: psychiatry and phenomenology today. AB - The paper illustrates the present role of phenomenological psychopathology by outlining its methods and focusing on some of its major issues: embodiment or body scheme, intentionality, time consciousness and intersubjectivity. The application of these categories to the analysis of psychotic disorders is demonstrated. Special emphasis is given to the relationship between phenomenology and cognitive neuroscience, a relationship that has been variously viewed as 'mutual constraint' or as 'mutual enlightenment'. This is where a new cooperation linking phenomenology, psychopathology and cognitive science begins to emerge. Phenomenology offers a methodically developed theory of human subjectivity that is indispensable to any attempt to understand, explore and treat psychiatric disorders. PMID- 12590190 TI - Typus melancholicus personality type and the five-factor model of personality. AB - Personality traits are significant factors in the development and course of depression. Apart from the classical five-factor model of personality, other personality constellations, such as Tellenbach's Typus melancholicus, have been described in association with depressive disorder. Several instruments have been developed to assess the Typus melancholicus personality (TMP). A systematic comparison of these instruments has not been done to date. The goal of this study was the comparison of four questionnaires used in assessing TMP. Four TMP questionnaires were compared and their relationship to the five-factor model of personality was examined among 264 psychiatric patients and normal controls. It was found that the TMP type represents a trait distinct from those of the five factor model. TMP inventories had only moderate concurrent validity. The single TMP scales focus on different aspects of the TMP, despite their common core. Both the five-factor personality traits and the TMP scales were able to differentiate a group of depressed patients from control groups. The results show that TMP is not one trait but a personality trait constellation. This leads to the conclusion that a number of dimensions must be taken into consideration in the construction of a TMP inventory. This multidimensionality contributes to the refinement of the TMP concept and differentiates its therapeutic implications. PMID- 12590191 TI - Autobiographical memory in major depression: a comparison between first-episode and recurrent patients. AB - Autobiographical memory in depression is characterized by an increase in general memory evocation. The aim of this study is to compare autobiographical memory in patients with a first depressive episode and in recurrent patients before and after recovery, using Williams' and Scott's autobiographical memory test. Our results show an increase of the number of general memories only with positive cue words in both groups of patients during the depressive episode. After clinical improvement, this specificity remains in recurrent patients who, in addition, recall more general memories for negative words. By contrast, patients with a first depressive episode are no longer different from controls. These results show both an overgeneralization and a deficit in positive memory access during the depressive episode, whatever the number of previous episodes. Moreover, recurrence chronically modifies access to emotional memories. PMID- 12590192 TI - Insight, social knowledge and working memory in schizophrenia. AB - BACKGROUND: There is evidence that insight and social judgements are impaired in schizophrenia. The influence of these factors on the decision to treat compulsorily in schizophrenia is poorly understood. AIMS: To investigate the contribution of insight, social knowledge, and working memory to the determination to treat coercively in schizophrenia. METHOD: Insight rating scale, social knowledge questionnaire and working memory tests were administered to detained patients with schizophrenia. Results were compared with those of a control group of voluntary in-patients with schizophrenia. RESULTS: Detained patients scored worse on insight and social knowledge, yet there was no significant correlation between these scores. There was no significant difference in severity of psychopathology between the experimental and control groups. Results for working memory were inconclusive. CONCLUSION: Insight and social knowledge are significantly, but independently, associated with the determination to treat coercively in schizophrenia. This suggests that insight and social knowledge are distinct skills. The contribution of working memory remains unclear. PMID- 12590193 TI - Attachment and panic disorder. AB - A dysfunctional relationship between parents and children can influence cognitive and emotional development and contribute to the development of psychiatric disorders, particularly panic disorder (PD). With the aim of exploring childhood experiences of parenting in PD patients, we compared subjectively perceived climate and objective recall by administering the Parental Bonding Instrument and 10 adjunctive items to 22 out-patients and 22 matched controls. Our analysis showed that DSM-III-R-diagnosed PD patients reported their parents to be significantly less caring than did the control group, while there was no significant difference in objective recall of parenting experiences. PMID- 12590194 TI - Violent women, blame attribution, crime, and personality. AB - BACKGROUND: The attribution of blame for criminal act has an impact, both on the way the offender must come to terms with the offence and on the risk of recidivism. Blame attribution has been found to be related to the type of crime and personality. AIMS: This study examined the relationship of blame attribution and feelings of guilt in violent female offenders, according to factors related to crime, background and personality. METHODS: The Gudjonsson Blame Attribution Inventory (BAI) and the Defense Style Questionnaire (DSQ) were administered to 58 violent female offenders. The Hare Psychopathy Checklist-Revised (PCL-R), Structured Clinical Interview II for DSM-IV (SCID II) and clinical interviews were used for assessment. RESULTS: Lower scores of guilt feelings were found in women than has been reported for men. The feelings of guilt were higher in homicide than in other violent crimes, but lower in the cases where women had committed prior violent crimes. No differences were found between psychiatric patients and prisoners. The duration of hospitalisation and incarceration did not result in differences in blame attribution and guilt, but when the victims survived, guilt feelings seemed to increase with the passing of the time. External attribution was related to motives of crime, but traumatic experiences did not have a crucial impact upon blame attribution. The attribution of blame and the amount of guilt feelings were found to be related to personality. Women scoring high on psychopathy reported less guilt than women with low scores. The corresponding result was not obtained in the group of women with antisocial personality disorder, who, however, showed an unexpectedly low use of external attribution. CONCLUSION: The results suggest that it might be difficult for women to cope with the violent offence and that the characteristics of psychopathy seems to have a crucial impact on the guilt feelings and consequently on coping with the crime in female offenders. PMID- 12590195 TI - 'Brain fag' symptoms in apprentices in Nigeria. AB - The 'brain fag' syndrome, which was first reported from West Africa in 1960 among students, has been shown to occur very widely in African students in western educational systems south of the Sahara. This study investigated the distribution of its symptoms in a group of apprentices chosen by convenient sampling from Ile Ife, a university town about 240 km northeast of Lagos in Nigeria. The subjects who (in contrast) were training under an indigenous form of education - the African apprenticeship system - consisted of 183 (69.8%) males, and 79 (30.2%) females, in the age range of 13-26 years (mean +/- SD 18.2 +/- 3.0 years). Questionnaires were interview-administered to collect data on the sociodemographic, economic, and family background, English language proficiency, and the degree of the presence of brain fag symptoms. Results indicated a generally low rate of brain fag symptoms among these different types of apprentices. A relationship between proficiency in English, but not socioeconomic status, with brain fag symptoms was found. In the case of the Nigerian apprentices investigated here, brain fag symptoms were not significantly associated with the method of training learning (which is dependent mainly on verbal instructions in the vernacular from their bosses, and vicarious learning by observing the boss at work). The implications of these findings for two of the theories advanced for the pathogenesis of the brain fag syndrome were discussed. PMID- 12590196 TI - Subjective cognitive dysfunction in first-episode patients predicts symptomatic outcome: a replication. PMID- 12590197 TI - Multifactorial determination of hypertensive nephroangiosclerosis. AB - Essential hypertension causes renal injury. Hypertensive nephroangiosclerosis (HN) or hypertensive nephropathy are terms most commonly used to describe this renal pathology. Although specific histological lesions occurring in affected kidneys are well known, pathogenesis of hypertension-related renal scarring is not completely understood. Evidence exists to support the theory that other factors such as aging, black race or smoking, beside blood pressure, contribute to the development and progression of HN. Metabolic disturbances, cocaine and nonsteroidal anti-inflammatory drug abuse, ochratoxin A exposure, dietary salt intake, heavy metal toxicity, hantavirus infection and perinatal programming are also considered risk factors. Renal susceptibility genes may determine whether hypertension-induced progressive renal damage occurs and how severe it is. Determination of all risk factors may identify patients at high risk of renal failure and help tailor an appropriate management. In the present paper, the knowledge available on this clinically important objective is discussed. PMID- 12590198 TI - Novel NCCT gene mutations as a cause of Gitelman's syndrome and a systematic review of mutant and polymorphic NCCT alleles. AB - BACKGROUND: Gitelman's syndrome (GS) is characterized by hypokalemic metabolic alkalosis, hypomagnesemia and hypocalciuria and these phenotypic features have been shown to be attributable to mutations in the gene encoding the thiazide sensitive Na/Cl cotransporter (NCCT). Until now, 55 different mutations have been reported and most of the families affected with GS exhibit autosomal recessive inheritance. METHODS: All 26 exons of the human NCCT gene were investigated in 2 German NCCT-deficient patients and their families. Mutation detection was performed by either direct automated sequencing of polymerase chain reaction (PCR)-amplified DNA products or by sequence analysis of cloned PCR products. RESULTS: In a 47-year-old German GS female a novel non-conservative missense mutation (S314F) and a complex deletion/insertion in the NCCT gene were found to be associated with the disorder. A further novel non-conservative substitution (S402F) together with a frequently observed R209W exchange were found in a 19 year-old German GS female. CONCLUSIONS: The observation of a compound heterozygote state in both females affected and the absence of a GS phenotype in their relatives carrying a single mutant allele is consistent with an autosomal recessive pattern of inheritance. PMID- 12590199 TI - Role of nitric oxide in the early renal changes induced by high fructose diet in rats. AB - BACKGROUND: Early glomerular hypertrophy and late glomerulosclerosis have been observed in rats fed high fructose diet (HFD), comparable with those of diabetic rats. Several studies suggest a role for nitric oxide (NO) in the pathogenesis of renal damage in diabetes. This study investigated the possible role of NO in the pathogenesis of HFD-induced glomerular changes. METHODS: Three study protocols were adopted. In the first, 20 rats were divided into two groups to evaluate the effect of HFD on glomerular size and on the urinary excretion of NO. In the second, the glomerular size was evaluated in 40 rats divided into four groups receiving: (1) standard diet (SD); (2) HFD; (3) HFD + L-NAME, and (4) SD + L-NAME for 1 month. In the third, the renal expression of inducible enzyme (iNOS) was compared in 10 rats on HFD and in 10 controls after a 1-month diet. RESULTS: The results showed: (1) increased urinary excretion of NO and glomerular size, both induced by HFD; (2) prevention by L-NAME of the HFD-increased glomerular size, and (3) increased iNOS expression in the kidneys of rats fed HFD. CONCLUSION: These results suggest a role for NO in the pathogenesis of the early renal changes induced by HFD. PMID- 12590200 TI - Activation of Na+/K+-ATPase by the serum and glucocorticoid-dependent kinase isoforms. AB - BACKGROUND/AIM: Expression of the constitutively active form of serum and glucocorticoid-dependent kinase ((S422D)SGK1) in Xenopus oocytes has recently been shown to upregulate endogenous Na(+)/K(+)-ATPase activity, an effect presumably participating in the regulation of cellular K(+) uptake and transepithelial Na(+) transport. SGK1 and the two isoforms SGK2 and SGK3 are stimulated by insulin and insulin-like growth factor-1 (IGF-1), which have been shown to enhance Na(+)/K(+)-ATPase activity in a variety of cells. The present experiments have been performed to elucidate whether or not wild-type SGK1, SGK2 and SGK3 are similar to (S422D)SGK1 in being effective regulators of Na(+)/K(+) ATPase. METHODS: To this end, dual-electrode voltage clamp experiments were performed in Xenopus oocytes injected either with water or with mRNA of constitutively active (S422D)SGK1 and wild-type SGK1, SGK2 or SGK3. Na(+)/K(+) ATPase activity was estimated from the outward-directed current created by readdition of extracellular K(+) in the presence of K(+) channel blocker Ba(2+) following a 10-min exposure to K(+)-free extracellular fluid. RESULTS: The outward-directed current was fully abolished by incubation with 1 mM ouabain and was significantly larger in oocytes expressing (S422D)SGK1, SGK1, SGK2 or SGK3, as compared to those injected with water. CONCLUSION: The stimulating effect of SGK1 on the Xenopus oocyte Na(+)/K(+)-ATPase is mimicked by the isoforms SGK2 and SGK3. Thus, all three kinases may participate in the regulation of Na(+)/K(+) ATPase activity by hormones such as insulin and IGF-1. PMID- 12590201 TI - The water-soluble fraction (<10 kD) of bee venom (Apis mellifera) produces inhibitory effect on apical transporters in renal proximal tubule cells. AB - Human envenomation caused by bee stings has been reported to cause acute renal failure and the pathogenetic mechanisms of these renal functional changes are still unclear. Bee venom is also a complex mixture of enzymes and proteins. Thus, this study was conducted to examine the effects of bee venom (BV, Apis mellifera) fractions on apical transporters' activity and its related signal pathways in primary cultured renal proximal tubule cells. Whole BV was extracted into three fractions according to solubility [a water-soluble fraction (BVA), an ethylacetate-soluble fraction (BVE), and a hexane-soluble fraction (BVH)]. BVA fraction was further separated to three portions according to molecular weights: BF1 (>20 kD), BF2 (10-20 kD), and BF3 (<10 kD). Each fraction was treated to the PTCs to the ratio of BV (1 microg/ml). BVA (930 ng/ml) significantly decreased cell viability, but BVH (27 ng/ml) and BVE (43 ng/ml) did not. BF3 (710 ng/ml) among BVA fractions predominantly decreased cell viability and inhibited alpha methyl-D-glucopyranoside (alpha-MG), phosphate (Pi), and Na(+) uptake. In addition, BF3 increased [(3)H] arachidonic acid release, lipid peroxide formation, and Ca(2+) uptake. These effects of BF3 were blocked by mepacrine and AACOCF(3) (phospholipase A(2) inhibitors) or N-acetylcysteine, vitamin C, and vitamin E (antioxidants). In conclusion, BF3 (<10 kD) among BV fractions is the most effective portion in BV-induced inhibition of alpha-MG, P(i), and Na(+) uptake and these effects of BF3 are associated with phospholipase A(2)-oxidative stress-Ca(2+) signal cascade in the primary cultured rabbit renal proximal tubule cells. PMID- 12590202 TI - Eluate derived by extracorporal antibody-based immunoadsorption elevates the cytosolic Ca2+ concentration in podocytes via B2 kinin receptors. AB - BACKGROUND/AIM: Patients with idiopathic focal segmental glomerulosclerosis (FSGS) often develop a recurrence of the disease after kidney transplantation. In a number of FSGS patients, plasmapheresis and immunoadsorption procedures have been shown to transiently reduce proteinuria and are thought to do this by eliminating a circulating factor. Direct cellular effects of eluates from immunoadsorption procedures on podocytes, the primary target of injury in FSGS, have not yet been reported. METHODS: Eluates were derived from antibody-based immunoadsorption of a patient suffering from primary FSGS, a patient with systemic lupus erythematosus, and a healthy volunteer. The cytosolic free Ca(2+) concentration ([Ca(2+)](i)) of differentiated podocytes was measured by single cell fura-2 microfluorescence measurements. Free and total immunoreactive kinin levels were measured by radioimmunoassay. RESULTS: FSGS eluates increased the [Ca(2+)](i) levels concentration dependently (EC(50) 0.14 mg/ml; n = 3-19). 1 mg/ml eluate increased the [Ca(2+)](i) values reversibly from 82 +/- 12 to 1,462 +/- 370 nmol/l, and then they returned back to 100 +/- 16 nmol/l (n = 19). The eluate-induced increase of [Ca(2+)](i) consisted of an initial Ca(2+) peak followed by a Ca(2+) plateau which depended on the extracellular Ca(2+) concentration. The eluate-induced increase of [Ca(2+)](i) was inhibited by the specific B(2) kinin receptor antagonist Hoe 140 in a concentration-dependent manner (IC(50) 2.47 nmol/l). In addition, prior repetitive application of bradykinin desensitized the effect of eluate on [Ca(2+)](i). A colonic epithelial cell line not reacting to bradykinin did not respond to eluate either (n = 6). Similar to FSGS eluates, the eluate preparations of both the systemic lupus patient and the healthy volunteer led to a biphasic, concentration-dependent [Ca(2+)](i) increase in podocytes which again was inhibited by Hoe 140. Free kinins were detected in all eluate preparations. CONCLUSION: The procedure of antibody-based immunoadsorption leads to kinin in the eluate which elevates the [Ca(2+)](i) level of podocytes via B(2) kinin receptors. PMID- 12590203 TI - A prospective randomized comparison of coralline hydroxyapatite with autograft in cervical interbody fusion. AB - STUDY DESIGN: A prospective randomized trial with independent clinical and radiographic outcome review of patients receiving either hydroxyapatite or tricortical iliac crest graft for cervical interbody fusion was conducted. OBJECTIVE: To determine whether coralline-derived hydroxyapatite is a suitable bone graft substitute in cervical interbody fusion. SUMMARY OF BACKGROUND DATA: Tricortical iliac crest bone is the "gold standard" graft material for cervical interbody fusion. Various bone substitutes have been used for this procedure to avoid potential donor site morbidity. ProOsteon 200 is a coralline-derived hydroxyapatite product, the use of which remains unclear for cervical interbody fusion. METHODS: In this study, 29 patients undergoing anterior cervical fusion and plating were randomized to receive either ProOsteon 200 or iliac crest grafts. The SF-36 and Oswestry Disability Index were used to measure clinical outcome. Postoperative radiographs were analyzed for graft fragmentation, loss of height, angular alignment, and hardware failure to assess structural integrity of the graft material. Plain radiographs and computed tomography scans were used to evaluate fusion. RESULTS: Both the ProOsteon 200 and iliac crest groups demonstrated significant improvement in clinical outcome scores. There was no significant difference in clinical outcome or fusion rates between the two groups. Graft fragmentation occurred in 89% of the hydroxyapatite grafts and 11% of the autografts (P = 0.001). Significant graft settling occurred in 50% of the hydroxyapatite grafts, as compared with 11% of the autografts (P = 0. 009). One patient in the ProOsteon 200 group required revision surgery for graft failure. CONCLUSIONS: ProOsteon 200 does not possess adequate structural integrity to resist axial loading and maintain disc height or segmental lordosis during cervical interbody fusion. PMID- 12590204 TI - Paraspinal muscle denervation, paradoxically good lumbar endurance, and an abnormal flexion-extension cycle in lumbar spinal stenosis. AB - STUDY DESIGN: A descriptive study was conducted to investigate the paraspinal muscle function in patients with lumbar spinal stenosis. OBJECTIVE: To evaluate paraspinal muscle innervation and endurance in lumbar spinal stenosis. SUMMARY OF BACKGROUND DATA: Abnormal lumbar paraspinal muscle function is associated with chronic low back pain. Abnormal neurophysiologic findings of the lower limbs often are observed in lumbar spinal stenosis, and abnormal lumbar function also can be expected. However, paraspinal muscle function and innervation have not been studied in lumbar spinal stenosis. METHODS: The study evaluated 25 patients with clinically and radiologically diagnosed lumbar spinal stenosis. Electromyography of the paraspinal muscles was performed from L3 to S1 bilaterally using a concentric needle. At least 20 insertions were analyzed from each muscle. The aim of the examination was to detect abnormal spontaneous activity associated with axonal damage (fibrillation potentials, positive sharp waves, and complex repetitive discharges). Paraspinal muscle activity during trunk flexion-extension movement and muscle endurance during the dynamic isoinertial back endurance test were assessed by surface electromyography. Muscle fatigue was calculated using mean power frequency analysis. RESULTS: Abnormal findings in needle electromyography of the paraspinal muscles were observed in 18 of the 22 (81.8%) examined patients. Abnormal flexion-extension activation of the paraspinal muscles was observed in all the examined patients. The change in mean power frequency was significantly smaller than in previously evaluated healthy subjects and patients with nonspecific chronic low back pain (P < 0.001) who were not experiencing symptoms of lumbar spinal stenosis. Paraspinal muscle fatigability was not associated with the denervation of the muscles. CONCLUSIONS Denervation and abnormal activation of lumbar paraspinal muscles are frequent findings in patients with lumbar spinal stenosis who have not undergone surgery. The paraspinal muscle endurance of the patients was unexpectedly good. PMID- 12590205 TI - Analysis of porous ingrowth in intervertebral disc prostheses: a nonhuman primate model. AB - STUDY DESIGN: A study was conducted to investigate the biomechanical, histochemical, and biologic ingrowth characteristics of the most widely used total disc prosthesis, the hydroxyapatite-coated SB Charite prosthesis. OBJECTIVE: To compare the porous ingrowth, linear apposition, or bony ingrowth in total disc replacement with published reports of porous ingrowth prostheses in the appendicular skeleton. METHODS: Seven mature baboons (Papio cynocephalus) underwent L5-L6 total disc replacement through an anterior transperitoneal approach. The SB Charite prosthetic vertebral endplates (n = 14) were cobalt chrome covered by two layers of thin titanium with a hydroxyapatite coating, which was electrochemically bonded to the implant surface. RESULTS: At 6 months after surgery, the range of motion exhibited by the SB Charite and the nonoperative control subjects under axial compression, flexion-extension, and lateral bending showed no statistical difference (P > 0.05). Plain film radiographic analysis showed no lucencies or loosening of any prosthetic vertebral endplate. Gross histopathologic analysis of the hydroxyapatite-coated SB Charite prosthesis demonstrated excellent ingrowth at the level of the implant bone interface, without evidence of fibrous tissue or synovium. Histochemical assays showed no accumulation of particulate wear debris (no titanium, ultrahigh molecular weight polyethylene, or cobalt-chrome) nor cytokines (tumor necrosis factor-alpha, prostaglandin E2, interleukin-1, -2, or -6). Total endplate area showed a mean ingrowth (volume fraction) of 47.9% +/- 9.12% and a total ingrowth range of 35.5% to 58.8%. CONCLUSIONS: The porous ingrowth (percentage of pore ingrowth coverage at the bone-metal interface) was more favorable for total disc replacement than for cementless total joint components in the appendicular skeleton (range, 10-30%). The reason for the improved degree of porous ingrowth in total disc replacement prostheses probably is that ligamentotaxis causes sustained compression across the metal-bone interface. PMID- 12590208 TI - Surgery of the lumbar spine for spinal stenosis in 118 patients 70 years of age or older. AB - STUDY DESIGN: A consecutive case retrospective chart review and an outcome satisfaction questionnaire were used in this study. OBJECTIVE: To provide a surgical reference for surgeons and elderly patients who may have concerns regarding the safety and outcome of lumbar spine surgery in their age population. SUMMARY OF BACKGROUND DATA: Elderly patients scheduled for spine surgery have a major concern about the safety and outcome of the procedure in light of their advanced age. A review of the literature demonstrated conflicting results regarding the outcome of lumbar spine surgery for spinal stenosis in the elderly. METHODS: A retrospective review evaluated 118 consecutive patients ages 70 to 101 years who were managed surgically for lumbar spinal stenosis. This patient population was analyzed for the operative procedure, postoperative morbidity and mortality, and long-term clinical outcome and satisfaction. All 118 patients had at least a 2-year follow-up evaluation, and 21 of these patients were older than 80 years. Clinical parameters were compiled and analyzed on the basis of chart review. RESULTS: Overall morbidity occurred in 24 patients (20%). During the study period, the average length of hospitalization declined an average of 2 days. Of the 118 patients, 109 expressed satisfaction with the operation and resumed daily activities, whereas 9 had fair or poor results. CONCLUSIONS: Advanced age did not increase the morbidity associated with this operation because the results reported in this study are comparable with those from other studies of a younger population, nor did advanced age decrease patient satisfaction or return to activities. PMID- 12590206 TI - Gabapentin effect on neuropathic pain compared among patients with spinal cord injury and different durations of symptoms. AB - STUDY DESIGN: This study evaluated the effect of gabapentin on neuropathic pain in patients with spinal cord injury. OBJECTIVE: To compare the effect of gabapentin on neuropathic pain refractory to conventional analgesics in patients with spinal cord injury and different durations of symptoms. SUMMARY OF BACKGROUND DATA: Neuropathic pain in patients with spinal cord injury severely compromises their quality of life. Gabapentin is a new antiepileptic drug that may additionally have a role in the treatment of neuropathic pain. So far, there has been little prospective research investigating the effect of gabapentin on neuropathic pain in patients after spinal cord injury or comparing gabapentin treated patients with varying durations of symptoms after spinal cord injury. METHODS: The study included 31 patients who had experienced neuropathic pain associated with spinal cord injury or cauda equina syndrome. These subjects were divided into two groups. Group 1 (n = 13) was composed of patients whose duration of pain was less than 6 months, and Group 2 (n = 18) comprised patients whose symptoms of neuropathic pain had lasted more than 6 months. Although these patients had been treated with conventional analgesics such as antidepressants, anticonvulsants, membrane stabilizer, and neuroleptics, they reported that their condition did not improve after a medication trial of at least 2 weeks duration. In this study, conventional analgesics were continued at a therapeutic level, and gabapentin was administrated for an 18-day titration period followed by a 5-week maintenance period at a dosage of 1800 mg/day or the maximum tolerable dosage. The efficacy of gabapentin administration was gauged by a pain score and a sleep interference score using a 100-mm visual analogue scale (VAS) every 2 weeks. The scores of the two groups were compared every 2 weeks over the course of the 8 week study. RESULTS: The mean pain score and the mean sleep interference score for Group 1 decreased more than that of Group 2 during the interval between 2 to 8 weeks (P < 0.05). The mean pain score for Group 1 decreased from 7.3 +/- 0.5 initially to 3 +/- 0.6 after 8 weeks of treatment, whereas the corresponding score for Group 2 decreased from 7.6 +/- 0.4 to 5.1 +/- 0.6 ( < 0.05). The mean sleep interference score for Group 1 decreased from 5.7 +/- 0.9 initially to 1.8 +/- 0.8 after 8 weeks of treatment, whereas the corresponding score for Group 2 decreased from 5.9 +/- 0.8 to 4.2 +/- 0.7 (P < 0.05). As compared with the onset of this study, a decrease in pain score of 2 or more was reported at the completion of this study for 11 patients (100%) in Group 1 and 10 (71%) of 14 patients in Group 2. A decrease of 2 or more in sleep interference scores was reported for 8 (89%) of 9 patients with sleep interference in Group 1 and for 8 (62%) of 13 patients with sleep interference in Group 2. Some adverse effects such as somnolence were noted, but they were mild or moderate in intensity. CONCLUSIONS: Gabapentin may be effective in decreasing neuropathic pain refractory to conventional analgesics in some patients with spinal cord injury whose duration of symptoms is less than 6 months, although those with duration of symptoms longer than 6 months showed a significant decrease as well. The drug is unlikely to cause serious adverse effects that limit its use in patients with spinal cord injury. PMID- 12590209 TI - Scott wiring for direct repair of lumbar spondylolysis. AB - STUDY DESIGN: This is a retrospective study of consecutive patients who underwent Scott wiring for symptomatic lumbar spondylolysis using the MacNab criteria. OBJECTIVE: The aim of the study was to assess the clinical outcome of the procedure after a mean follow-up period of more than 10 years. SUMMARY OF THE BACKGROUND DATA: Previous studies have shown this technique to be effective in the management of patients up to the age of 25 years who have symptomatic lumbar spondylolysis with or without a Grade 1 slip. No study has been published with regard to the long-term outcome of Scott wiring. METHODS: Between 1986 and 1993, Scott wiring was performed for 15 patients younger than 25 years of age who had symptomatic lumbar spondylolysis. MacNab criteria were used to assess their pre- and postoperative status. A postal questionnaire was used to assess their clinical outcome. Their notes also were studied to find subsequent episodes of low back pain. RESULTS: The postoperative questionnaires were returned by 14 patients. The mean follow-up period was 10.9 years. Of the 14 patients, 12 had a "good" or "excellent" result according to MacNab criteria. The two patients classified as "poor" required further procedures for continued low back pain. CONCLUSIONS: The Scott wiring technique is recommended for the treatment of symptomatic lumbar spondylolysis in patients younger than 25 years because it gave "good" or "excellent" results in approximately 86% of the patients with a mean follow-up period of 10.9 years. PMID- 12590210 TI - Results of surgical treatment of adult idiopathic scoliosis with low back pain and spinal stenosis: a study of long-term clinical radiographic outcomes. AB - STUDY DESIGN: A case series of adults with surgical treatment for adult idiopathic thoracolumbar and/or lumbar scoliosis, low back pain, and spinal stenosis was studied. OBJECTIVE: To assess pain relief, curve correction, and complications after combined procedures consisting of decompression, spine fusion, and stabilization. SUMMARY OF BACKGROUND DATA: Only one publication has focused specifically at this group, and this was before the advent of modern segmental instrumentation. This is the first report of long-term follow-up evaluation in such a patient population. METHODS: This study included 16 patients who underwent elective anterior and posterior surgical reconstruction for adult idiopathic thoracolumbar and/or lumbar scoliosis, spinal stenosis, and low back pain with a minimum follow-up period of 2 years. Radiographic findings, clinical results, and long-term outcome data were obtained using the Modified Scoliosis Research Society outcome instrument and the Oswestry Disability Back Pain Questionnaire. RESULTS: Restoration of coronal and sagittal balance, or improvement thereof, was achieved in all the patients with balance problems. There was significant improvement in all outcome domains. Overall, 94% of the patients were satisfied with the surgery. Ten major complications occurred in 10 patients, 8 of whom required additional surgery. There were two minor complications. CONCLUSIONS: Combined symptoms of back pain and spinal stenosis require complex reconstructive surgery in adults with idiopathic thoracolumbar and/or lumbar scoliosis. Significant pain relief, functional restoration, and satisfaction can be achieved and maintained over the long term in the properly selected patient. PMID- 12590211 TI - Buckling and bone modeling as factors in the development of idiopathic scoliosis. AB - STUDY DESIGN: Computational analysis using the finite-element method was used to examine a possible etiology of idiopathic scoliosis. OBJECTIVES: To compare changes in the coronal and the transverse planes of idiopathic thoracic scoliosis with changes produced in a finite-element buckling model, and to investigate the influence of bone modeling on the buckling spine. SUMMARY OF BACKGROUND DATA: Although it is now widely accepted that growth is related strongly to the onset and progression of scoliosis, the pathomechanism or etiology of idiopathic scoliosis still is not clear. A previous study showed that a buckling phenomenon caused by anterior spinal overgrowth can produce scoliosis, and that the fourth buckling mode matched the clinical characteristics associated with the thoracic type of idiopathic scoliosis. The fourth buckling mode occurs when the first, second, and third buckling modes are prevented. METHODS: The spinal finite element model used in this study consisted of 68,582 elements and 84,603 nodes. The transverse changes seen in the computed tomography images of 41 patients with idiopathic thoracic scoliosis (apex, T8; average Cobb angle, 52.5 degrees) were compared with those produced in the fourth buckling mode. Bone modeling (bone formation and resorption) was simulated as heat deformation caused by changes in temperature. The bone formation and resorption were simulated, respectively, by positive and negative volume changes in proportion to the stress that occurred in the buckling spine. RESULTS: Computed tomography images of scoliosis show that as the scoliosis becomes more severe, the thoracic cage decreases on the convex side of the curve and increases on the concave side. The opposite thoracic cage deformation was obtained in the fourth buckling mode. In patients with scoliosis, the sternum essentially remains in its original position with respect to the vertebrae, but in the linear buckling model, it shifted in the direction of vertebral body rotation. In contrast to clinical data, the incremental deformation resulting from bone formation corrected the original curve, and the thoracic cage distorted. On the other hand, incremental deformation resulting from bone resorption worsened the original curve, and the thoracic cage distorted in a manner similar to that described by the clinical data. CONCLUSIONS: This computational investigation suggests that scoliotic changes in the spinal column triggered by the buckling phenomenon are counteracted by bone formation, but worsened by bone resorption. The authors hypothesized that scoliosis progressed with resorption of loaded bone. However, it is unclear whether this hypothesis applies to a living body in practice because of the effects from additional factors. PMID- 12590213 TI - Radiographic assessment of interbody fusion using recombinant human bone morphogenetic protein type 2. AB - STUDY DESIGN: A prospective randomized study investigated the radiographic progress of fusion at 6, 12, and 24 months in 42 patients who underwent a single level anterior lumbar interbody fusion using cylindrical interbody fusion cages. OBJECTIVE: To determine the patterns and rates of osteoinduction associated with the use of recombinant human bone morphogenetic protein type 2 (rhBMP-2) and an absorbable collagen sponge carrier in anterior lumbar interbody fusion with a tapered cylindrical fusion device. SUMMARY OF BACKGROUND DATA: Studies have shown that rhBMP-2 used with allograft dowels increases the rate of interbody fusion by promoting osteoinduction and enhancing incorporation of the allograft. In a small series of human patients undergoing anterior lumbar interbody fusion with a tapered cylindrical fusion cage, rhBMP-2 has been shown to promote osteoinduction and fusion. METHODS: In this prospective nonblinded study, 42 patients were randomly divided into two groups. The investigational group underwent interbody fusion using two tapered cylindrical fusion cages (LT-CAGE) and rhBMP-2 on an absorbable collagen sponge, and a control group underwent the procedure, receiving the devices and autogenous iliac crest bone graft. Plain radiographs and computed tomographic scans were used to evaluate the pattern of osteoinduction in the interbody space and the progression of fusion 6, 12, and 24 months after surgery. RESULTS: All the patients who received rhBMP-2 showed radiographic evidence of osteoinduction in the interbody cages 6 months after surgery. The density in the cages had increased an average of 142 Hounsfield units by 6 months. At 12 months, the increase had reached 228.7 Hounsfield units New bone formation occurred in the disc space outside the cages by 6 months in 18 of the patients in the investigational group (18/22; 82%). By 24 months, all the investigational patients showed new formation outside the cages. In the autograft control group, the density in the cages increased an average of 42 Hounsfield units, and 10 patients (10/20; 50%) showed evidence of bone formation outside the cages. CONCLUSIONS: The use of rhBMP-2 is a promising method for facilitating anterior intervertebral spinal fusion in patients who have undergone anterior lumbar fusion surgery. PMID- 12590214 TI - Significance of bone formation markers in patients with ossification of the posterior longitudinal ligament of the spine. AB - STUDY DESIGN: Serum concentrations of bone formation markers were correlated with the type, location, and progression of ossification of the posterior longitudinal ligament. OBJECTIVE: To determine the relation between bone formation markers and ossification of the posterior longitudinal ligament. SUMMARY OF BACKGROUND DATA: Few reports have correlated bone formation markers with ossification of the posterior longitudinal ligament. METHODS: In this study, 43 patients with cervical ossification of the posterior longitudinal ligament and myelopathy underwent laminoplasty. The patients were observed for more than 10 years, after which plain radiographs and tomograms of the cervical region were taken. The radiographs were selectively performed to address thoracic and lumbar ossification of the posterior longitudinal ligament. Serum concentrations of bone formation markers (intact osteocalcin, osteocalcin, carboxyterminal propeptide of human type 1 procollagen, and bone-specific alkaline phosphatase) were measured and correlated with these radiographic studies. RESULTS: A positive correlation was observed between intact osteocalcin, osteocalcin, and carboxyterminal propeptide of human type 1 procollagen in patients with combinations of cervical, thoracic, or lumbar ossification of the posterior longitudinal ligament. CONCLUSIONS: Serum concentrations of intact osteocalcin, osteocalcin, and carboxyterminal propeptide of human type 1 procollagen may reflect the activity of general ectopic bone formation in patients with ossification of the posterior longitudinal ligament. PMID- 12590215 TI - A comparison of lumbar range of motion and functional ability scores in patients with low back pain: assessment for range of motion validity. AB - STUDY DESIGN: A clinical convenience sample of workers with low back problems was tested once for a variety of parameters. OBJECTIVES: To quantify the links between lumbar range of motion and scores obtained from functional evaluation tests. SUMMARY OF BACKGROUND DATA: Although lumbar range of motion traditionally has been the standard used to determine disability for the purposes of compensation, a concern arises from the fact that a discrete physical impairment associated with low back pain and diminished range of motion often is never found. METHODS: This study involved 18 workers with chronic low back pain referred to a rehabilitation center for determination of compensation and fitness for return to work. Lumbar range of motion was precisely and accurately measured with a three-dimensional lumbar motion instrument. In addition, typical functional tests were performed for each patient. Comparisons were made between all the lumbar range of motion values and each of the functional ability scores. RESULTS: The most notable finding was the lack of significant correlation between most of the lumbar range of motion values and the functional test scores. Only 9 of a possible 144 correlations were significant. CONCLUSIONS: The relation between lumbar range of motion measures and functional ability is weak or nonexistent. This has implications for clinical practice as it relates to disability determination for patients with chronic low back pain, and perhaps for the current impairment guidelines of the American Medical Association. PMID- 12590216 TI - Gender differences in sick leave related to back pain diagnoses: influence of pregnancy. AB - STUDY DESIGN: A cross-sectional population-based study was conducted. OBJECTIVE: To investigate the influence of pregnancy on any gender differences in sickness absence related to back pain diagnoses in the group 16 to 44 years of age. SUMMARY OF BACKGROUND DATA: Back pain diagnoses are a major reason for sickness absence, especially among pregnant women. There is, however, little knowledge on the extent to which back pain during pregnancy may account for any differences in sickness absence when compared with men and nonpregnant women. METHODS: Data on sick leave periods certified by a physician were combined from two research databases: one including all insured persons (n = 50,167) and one including pregnant women (n = 1342) for the duration of 1 year. RESULTS: When pregnant women were subtracted from "all women," nonpregnant women had the same cumulative incidence of sickness absence related to back pain diagnoses as men. Among "all women" sick-listed with back pain diagnoses, 37% were pregnant, a finding not evident from official statistics. Sickness absence increased with age among nonpregnant women and men, but decreased among pregnant women. CONCLUSIONS: The proportion of sickness absence related to back pain diagnoses did not differ between nonpregnant women and men ages 16 to 44 years. The importance of back pain diagnosis as a reason for sickness absence may thus be overrated among nonpregnant women. PMID- 12590217 TI - The relation of race to outcomes and the use of health care services for acute low back pain. AB - STUDY DESIGN: Four strata of randomly selected health care providers in North Carolina (primary care MDs, Doctors of Chiropractic, orthopedic surgeons, and group model HMO primary care providers) enrolled 1633 consecutive patients with low back pain into a cohort study. OBJECTIVE: To determine whether race had an independent effect on rate of recovery from low back pain, and whether there was any racial disparity in the treatments provided to patients with low back pain. SUMMARY OF BACKGROUND DATA: Little research to date has examined the relation between patient race and recovery from an episode of acute low back pain. METHODS: Consecutive patients were enrolled in the provider's office and contacted by telephone at baseline, at 2, 4, 8, 12, and 24 weeks, and at 22 months. RESULTS: Blacks (n = 238) at baseline had higher pain scores on a 10 point scale (5.92 vs 5.25; P< 0.01) and worse functional disability (12.1 vs 11; P= 0.04), as assessed by the 23-point Roland-Morris scale, yet were considered by their health provider as having less severe pain and less likely to have disc disease than white patients (P < 0.05 for all comparisons). Blacks had worse functional disability at most follow-up interviews. Blacks were shown to be less likely to receive radiographs (49% vs 40%) or advanced imaging studies (10% vs 6%), even after controlling for income, education, baseline severity of low back pain, and insurance status (P < 0.05). Doctors of Chiropractic had different practice approaches than MDs, and there was an interaction with patient race. CONCLUSIONS: The relation of patient race to outcomes from and care for low back pain is complex. Blacks have slightly worse functional status than whites on presentation and at follow-up assessment. Blacks receive less intense diagnostic and treatment approaches from MDs, although the severity of their impairment is at least as great. PMID- 12590218 TI - Back pain online: a cross-sectional survey of the quality of web-based information on low back pain. AB - STUDY DESIGN: A cross section of Web sites accessible to the general public was surveyed. OBJECTIVE: To evaluate the quality of information on low back pain and its treatment that a "typical" patient user might access on the Internet. SUMMARY OF BACKGROUND DATA: Individuals with back pain have a desire to learn about their condition, what to expect, and what they can do about it. Web sites play a potentially useful role in providing information to help people learn about their low back pain and select the most appropriate methods of management. METHODS: A general search using popular search engines located 60 Web sites about back pain for review. A list of criteria for evaluating and scoring back pain Web sites was established using available literature and current clinical guidelines for the management of acute low back pain. The total quality score (maximum score, 38) was composed of two separate scores: one for general quality of the site (maximum score, 14) and one for site content specific to low back pain (maximum score, 24). Statistical tests, as appropriate, were used to investigate the relation between general indicators of Web site quality and total scores obtained. RESULTS: The quality of the Web sites surveyed was poor, most of them (n = 58, 97%) scoring less than half the maximum available score. The mean total score was 7.4 (range, 2-25). The mean score was 4.9 (range, 1-12) for general Web site quality and 2.4 (range, 1-13) for content specific to low back pain. Web sites providing references, sites created more recently, and sites not created for advertising purposes tended to be of better quality. CONCLUSIONS: This study highlighted the poor quality of information, particularly information about low back pain, available to "typical" patient users on the Internet. Health care professionals must have a role in evaluating existing information and in developing good-quality evidence-based Web sites. Patients with back pain should be discouraged from using the Internet as a source of information unless the Web sites they access have been evaluated and found to provide evidence-based messages. PMID- 12590219 TI - Accuracy of thoracic vertebral body screw placement using standard fluoroscopy, fluoroscopic image guidance, and computed tomographic image guidance: a cadaver study. AB - STUDY DESIGN: A surgical simulation study in human cadaver spine specimens was conducted to evaluate the accuracy of thoracic vertebral body screw placement using four different intraoperative imaging techniques. OBJECTIVE: To compare standard fluoroscopy, fluoroscopy-based image guidance with two different referencing methods, and computed tomography-based image guidance by the measuring the time required for screw placement, the radiation exposure to specimen and surgeon, and the accuracy of screw position in the thoracic spine. SUMMARY OF BACKGROUND DATA: Image guidance provides additional anatomic information to the surgeon and may improve safety of technically difficult surgical procedures. The placement of screws in the thoracic spine is a technically demanding procedure in which inaccurate screw positioning places the spinal cord, nerve roots, and paraspinal structures such as the aorta and pleural space at risk for injury. Image-guided surgery may improve the accuracy of thoracic screw placement. METHODS: Using four different intraoperative imaging methods, two experienced surgeons placed 337 vertebral body screws through the pedicles of thoracic vertebrae in 20 human cadaver thoracic spine specimens. The specimens then were examined with radiographs, computed tomography, and anatomic dissection to determine screw position. Measurements included procedure setup and screw insertion time, radiation exposure to the specimen, the surgeon's hand, the surgeon's body, frequency, direction, and magnitude of screw perforation through the cortical margins of thoracic vertebrae. RESULTS: As compared with surgery using standard fluoroscopy, fluoroscopy-based image guidance that uses multiple reference marks and computed tomography-based image guidance improves the accuracy of thoracic vertebral body screws, but increases the time required for screw placement and the specimen radiation exposure. Exposure to radiation is minimal at the surgeon's body level and dependent on surgical technique at the surgeon's hand level. Screw perforation occurs most frequently in the lateral direction. CONCLUSIONS: Fluoroscopy-based image guidance that uses only a single reference marker for the entire thoracic spine is highly inaccurate and unsafe. Systems with registration based on the instrumented vertebrae provide more accurate placement of thoracic vertebral body screws than standard fluoroscopy, but expose the patient to more radiation and require more time for screw insertion. PMID- 12590220 TI - Occipitocervical fusion for rheumatoid arthritis using the inside-outside stabilization technique. AB - STUDY DESIGN: A retrospective study investigating the clinical outcome of the inside-outside cranial bolt technique for occipitocervical stabilization used to manage rheumatoid arthritis was conducted. OBJECTIVE: To evaluate the safety and efficacy of the inside-outside technique for occipitocervical stabilization used to manage rheumatoid patients. SUMMARY OF BACKGROUND DATA: Achieving occipital cervical fusion for patients with rheumatoid arthritis is a complex and challenging problem. Complications related to placement of occipital screws have been reported. METHODS: Occipitocervical stabilization was used for atlantoaxial subluxation or basilar invagination in 21 patients with rheumatoid arthritis. The patients were assessed for pre- and postoperative neurologic status (Ranawat classification), neck pain, fusion and alignment, hardware complications, and continued posterior cranial settling. All the patients underwent stabilization using inside-outside occipital screws. The technique involves bilateral fixation of cervical plates to the occiput using inside-outside screws, and to the cervical spine using pars screws at C2 or lateral mass screws at subaxial levels. Bone grafting was accomplished with autologous rib or iliac crest graft. RESULTS: During the study, 14 women and 7 men with rheumatoid arthritis underwent occipitocervical stabilization and fusion. The average age of the patients was 65 years, and the mean follow-up period was 25.5 months. There were no surgical complications. Neck pain was reduced from an average Ranawat pain score of 2.40 to 0.4 (P < 0.0001). The Ranawat neurologic grade improved in 62% of the patients with preoperative neurologic deficit. Further cranial settling was not observed in any patient. There were no complications from implants and no incidence of instrumentation failure. CONCLUSIONS: The "inside-outside" technique is safe and effective for stabilizing the occipitocervical junction in rheumatoid patients. It is associated with significant reduction of neck pain, improved neurologic status, and maintenance of alignment and stability. PMID- 12590221 TI - Cervical spondyloptosis caused by an aneurysmal bone cyst: a case report. AB - STUDY DESIGN: The case of an 11-year-old patient with an aneurysmal bone cyst at C7, cervical spondyloptosis, and scoliosis of the lumbar spine is reported. OBJECTIVE: To describe an unusual presentation of aneurysmal bone cyst causing painless cervical spondyloptosis. SUMMARY OF BACKGROUND DATA: Aneurysmal bone cysts of the cervical spine are rare. There has never been a reported case of cervical dislocation by an aneurysmal bone cyst. This case highlights the potentially devastating effects that can be caused by benign tumors in the vertebral column. METHODS: The history, clinical examination, imaging findings, and treatment are reviewed. RESULTS: The patient was treated with a single-stage combined posteroanteroposterior operation to resect the tumor, reduce the cervicothoracic dislocation, and create a three-column stabilization of the spine. The results were satisfactory, with an improvement in neurology. CONCLUSIONS: Aneurysmal bone cysts should be considered in the etiology of complicated scoliosis. Patients with a benign clinical history should be investigated at an early stage using appropriate imaging. Surgery is the treatment of choice when stability of the spine is compromised and neurologic signs are present. PMID- 12590222 TI - Delayed diagnosis of a false aneurysm after lumbar arterial injury: treatment with endovascular embolization: a case report. AB - STUDY DESIGN: A case report is presented. OBJECTIVES: To report a case of lumbar artery pseudoaneurysm that was primarily missed, and to discuss therapeutic options. SUMMARY OF BACKGROUND DATA: In the literature, only a few cases of lumbar artery pseudoaneurysms have been reported. The diagnosis is difficult and often delayed, with a sometimes fatal outcome. The treatment is either surgery or endovascular embolization. METHODS: In a patient who survived after a fall from 20 feet (6 m) height, a relatively small retroperitoneal hematoma detected during urgent splenectomy was underestimated. Two weeks later, the underlying laceration of the lumbar artery led to the formation of a pseudoaneurysm, which then ruptured causing a large retroperitoneal hematoma and gradual complete femoral nerve palsy. RESULTS: Complete occlusion of the pseudoaneurysm and progressive regression of the retroperitoneal hematoma were achieved by two stages of endovascular embolization. The size of the hematoma was diminished gradually during a period of 12 months, whereas the femoral nerve has completely recovered. CONCLUSIONS: Traumatic rupture of a lumbar artery is a rare complication of a blunt abdominal trauma. This condition leads to a potentially massive retroperitoneal hemorrhage and shock, or to subsequent pseudoaneurysm formation and delayed retroperitoneal hematoma. This condition can be managed with either surgical or preferably endovascular intervention provided the patient is hemodynamically stable. PMID- 12590223 TI - Osteogenic sarcoma of the rib: a case presentation and literature review. AB - STUDY DESIGN: A case report is presented. OBJECTIVES: Primary tumors of the rib are relatively uncommon in the adult population, and even more rare in children. A case of osteogenic sarcoma of the rib and a literature review are presented. SUMMARY OF BACKGROUND DATA: Osteogenic sarcoma represents approximately 30% of all malignant sarcomas diagnosed in the United States. A single case of osteogenic sarcoma of the rib has been reported in the literature involving a 9 year-old child. METHODS: Clinical data analysis. RESULTS: A 9-year-old white boy presented with a mass of the left posterior thorax. The initial chest radiograph showed a nonhomogeneous mass with calcifications adjacent to the 11th rib. The final diagnosis was osteogenic sarcoma. Chemotherapy was initiated. The patient underwent radical excision of the mass. Given the extent of the patient's resection, it was thought that he would be at high risk for the development of spinal deformity. He was placed in a TLSO brace (Bolt Systems, Orlando, FL) and followed closely. At 15 months after excision of his tumor, he was noted to have progression of a thoracolumbar scoliosis and significant kyphosis. At this writing, it has been 52 months since resection. The patient has no evidence of local recurrence or metastatic disease, and his spinal curvature remains stable. CONCLUSIONS: The patient's short-term (4-year) disease-free survival illustrates the efficacy of neoadjuvant chemotherapy and radical surgical resection. Patients with osteogenic sarcoma of the rib should be monitored closely for the development of spinal deformity if the required resection includes the vertebral column. Casting and bracing may help to limit progression of the deformity. PMID- 12590224 TI - Thoracic spinal cord injury without radiographic abnormality in a skeletally mature patient: a case report. AB - STUDY DESIGN: A case of thoracic spinal cord injury without radiographic abnormality in a skeletally mature patient is reported. OBJECTIVES: To report, for the first time in the literature, a case of thoracic spinal cord injury without radiographic abnormality or any ligamentous injury in a skeletally mature patient, and to propose a unique mechanism of spinal cord traction injury resulting from associated bilateral high-energy lower limb injuries. SUMMERY OF BACKGROUND DATA: Thoracic spinal cord injury without radiographic abnormality is extremely uncommon in skeletally mature patients. Hyperflexion with an associated distraction force usually produces the spinal cord injury, which is associated with posterior ligamentous injury of the spine. In the only reported case of thoracic spinal cord injury without radiographic abnormality in a skeletally mature patient to date (1993), despite the absence of evidence showing bony injury associated with cord damage on plain radiographs and computed tomograms, the magnetic resonance image demonstrated significant injury to the posterior ligamentous structures of the spine. Thoracic spinal cord injury in a skeletally mature patient without associated bony and ligamentous injury has never before been reported. METHODS: A 17-year-old skeletally mature motorcyclist was involved in a road traffic accident and sustained multiple injuries. Clinical examination showed tenderness over L1 and L2 vertebrae with complete loss of sensation and motor power below L2. In addition, the patient also sustained a stable pelvic fracture; fracture of the right acetabulum; open dislocation of the right knee with complete disruption of both cruciate ligaments, the posteromedial capsule, and the medial collateral ligament; open fracture of the left tibia and fibula; displaced fracture of the medial condyle of the left femur; fracture of the right distal radius; and fracture of the right proximal humerus. The plain radiographs of the whole spine and computed tomography scanning of the thoracolumbar spine showed no evidence of bony injury. The contemporary magnetic resonance image scanning of the entire spine showed disruption of the spinal cord from T10 downward, with patchy high signal in the cord and loss of normal architecture. After appropriate management of the associated limb injuries, the patient was transferred to the regional spinal unit for rehabilitation. RESULTS: Thoracic spinal cord injury in a skeletally mature patient occurred in the absence of associated bony and ligamentous injury, and probably resulted from a longitudinal traction force transmitted through the sciatic nerves to the spinal cord as a result of associated high-energy bilateral lower limb injuries. CONCLUSIONS: A rare case of thoracic spinal cord injury without radiographic abnormality manifested as a result of traction injury to the sciatic nerves caused by bilateral violent lower limb injuries in a skeletally mature patient is presented. To the best of the authors' knowledge, such a case has not been reported previously. PMID- 12590225 TI - Lateral approach to a neurenteric cyst of the cervical spine: case presentation and review of surgical technique. AB - STUDY DESIGN: The lateral cervical spinal approach is illustrated by a case of neurenteric cyst. OBJECTIVES: To present a case of a neurenteric cyst resected the lateral cervical approach, and to review the approach in detail. SUMMARY OF BACKGROUND DATA: Intradural lesions located anterior to the high cervical spinal cord may present a difficult surgical problem. Neurenteric cysts are unusual lesions found in the brain and spinal cord. This report presents a case of cervical neurenteric cyst causing anterior cord compression that was resected using the lateral cervical approach. METHODS: A 32-year-old woman presented with chronic headaches and worsening nausea, tinnitus, dizziness, and hyperreflexia and clonus in her lower extremities. Magnetic resonance imaging of the cervical spine showed a mass compressing the anterior spinal cord at C3. Pathology showed that this lesion was a neurenteric cyst. The lesion was resected using the lateral cervical approach. RESULTS: At this writing, 36 months after surgery, the patient has continued resolution of her symptoms, and no cyst recurrence has been shown on repeat MRI imaging. She has no evidence of postlaminectomy kyphosis. CONCLUSIONS: The lateral cervical approach is useful for surgeons attempting to resect lateral and anterior intradural lesions of the cervical spine. It also gives excellent cranial-to-caudal access to the thecal sac, spinal cord, and the lesion to be resected. The neck incision preserves cosmesis, and neuromuscular function is maintained. Spinal fusion was avoided in the reported case. PMID- 12590226 TI - Optimal management of nonalcoholic fatty liver/steatohepatitis. PMID- 12590227 TI - Predicting severity in acute pancreatitis: in search of a perfect marker. PMID- 12590228 TI - Gastroesophageal reflux disease and the larynx. AB - Gastroesophageal reflux disease (GERD) is associated with ear, nose, and throat (ENT) signs and symptoms. Previously, experiments in animal larynx suggested that acid in combination with pepsin resulted in more laryngeal injury than acid alone. Recently, similar experiments expanded on the above observation, finding no laryngeal inflammation with bile acid, trypsin, or combination of bile constituents in alkaline pH values. Combined, experimental evidence highlights the importance of acidic refluxates that contain pepsin and/or bile acids; however, many patients with laryngeal signs suggestive of GERD do not respond to aggressive acid suppression. This has resulted in controversy about the exact nature of relationship between GERD and ENT signs and symptoms. Because the diagnosis is initially suspected after a laryngoscopic examination by the ENT physicians, the suboptimal clinical response may be caused by the poor specificity of this exam in identifying GERD as the cause of patients' laryngeal symptoms. Hence, a multidisciplinary approach is often required in the optimum treatment of these patients. PMID- 12590229 TI - Family history of gastric cancer: should we test and treat for Helicobacter pylori? AB - A close link has been established between infection and gastric cancer. In this article, we suggest that using a risk stratification technique (like that for colorectal cancer), the high-risk group of first-degree relatives of patients with gastric cancer can be separated out for testing and treatment. This would be more manageable and more cost-effective than screening the whole population, in which the mortality from distal gastric cancer has declined concomitant with the eradication of infection. Support for the feasibility of this approach is derived from studies showing that the family is the core unit of transmission and that childhood colonization, especially with a virulent strain, is apparently a major risk factor for disease progression to the neoplastic stage. When there is a case of gastric cancer in the family, first-degree relatives, who might be infected by a bacterium with an identical genetic fingerprint, are at higher risk than normal for developing gastric cancer. Furthermore, genetic and epidemiologic studies based on the Correa model have shown that both primary and secondary prevention of gastric cancer is possible. Calculations done in high-risk populations, such as Japanese-Americans, confirm the savings in cost and the safety of the test-and treat strategy. Considering that eradication should be done as early as possible, at a point in the cascade when the changes are still reversible, and that gastric cancer is associated with a high mortality rate, we suggest that this strategy be applied to this high-risk population. PMID- 12590230 TI - Intrasphincteric botulinum toxin versus pneumatic balloon dilation for treatment of primary achalasia. AB - GOALS: Despite a high success rate, pneumatic dilatation for achalasia is accompanied by a significant risk of esophageal perforation. Injection of botulinum toxin (botox) into the lower esophageal sphincter (LES) can lead to improvement in symptoms with reduced risk of complications. Direct comparisons of the two techniques are needed to define their role in clinical management. STUDY: We compared pneumatic dilatation to botox for patients with achalasia using a double blind, randomized study design. Patients underwent clinical, manometric, radiographic and endoscopic evaluation to confirm primary achalasia. They were randomized to receive either 80 units of botox into the LES or Witzel balloon dilatation. Patients also received sham dilatation or injection, respectively. The patients and investigators assessing symptom response were blinded to therapy. Symptoms and esophageal function were assessed at 3 weeks, 3 months and 1 year after therapy. Treatment failure was defined as the lack of decrease in symptom grade more than 1 or recurrence of symptoms. Patients with treatment failure crossed over to the alternative treatment. RESULTS: Thirty four patients were studied, and 31 completed the trial. Of the 18 patients randomized to Witzel dilatation, 16 (89%) of 18 remained in clinical remission. Of the two patients with treatment failure, one responded to botox injection. Of the 16 patients randomized to botox, (38%) 6 of 16 remained in clinical remission. Four patients had initial failure, and 6 relapsed at a mean of 4 months after therapy. Of the nine patients who crossed over to dilatation, seven responded well, but two required surgical management of perforation. Although both treatments had excellent initial clinical improvement, patients randomized to Witzel dilatation had superior long-term success ( < 0.01). CONCLUSION: Initial therapy with Witzel dilatation is associated with better long-term outcome than a single injection of botox. Because of the risk of endoscopic perforation, botox remains a viable alternative to dilatation. PMID- 12590231 TI - Thermal heterogeneity constitutes a marker for the detection of malignant gastric lesions in vivo. AB - BACKGROUND: Several studies have attempted to investigate the association of thermal difference between malignant tumors and inflammatory benign lesions. In this work, we evaluated whether thermal heterogeneity of solid tumors in the stomach constitutes a marker for the diagnosis of benign, preneoplastic, and malignant lesions. STUDY: A thermistor probe was used that attached to the distal end of a long and steerable, 3fr-thermography catheter passed through the biopsy channel of the gastroscope and came in touch with the stomach epithelium to measure temperature differences (DeltaT) between normal tissue and various types of lesions. The method was applied in 8 patients with benign hyperplastic gastric lesions, 19 patients with gastritis, 9 patients with peptic ulcer, 7 patients with dysplasia and 11 patients with gastric adenocarcinoma. RESULTS: Progressive changes in DeltaT between hyperplastic gastric lesions, gastritis, ulcer, dysplasia and adenocarcinoma were observed (P < 0.001). Statistical analysis showed that DeltaT greater than 1.7 degrees C, constitutes a crucial point for the diagnosis of malignancy, in stomach lesions, with sensitivity (72%) and specificity (94%). CONCLUSIONS: These findings suggest that the detection of DeltaT, between normal tissue and neoplastic lesions, could be useful in clinical practice for the differential diagnosis of stomach lesions, even in the early stages. PMID- 12590232 TI - Prevalence of antitissue transglutaminase antibodies in different degrees of intestinal damage in celiac disease. AB - GOALS: Although anti-tissue transglutaminase antibodies (anti-tTG) are effective for celiac disease (CD) routine laboratory screening, there are no studies evaluating correlation between degree of intestinal damage and positivity to anti tTG. Since recent studies showed that anti-gliadin (AGA) and anti-endomysium (EMA) antibodies are ineffective in diagnosing mild gluten-sensitive enteropathy, the aim of this study was to evaluate the prevalence of anti-tTG in different degrees of intestinal damage of celiac patients and whether there is a correlation between serum value of anti-tTG and the degree of histologic damage. STUDY: We studied 119 consecutive adult patients affected by CD (47 men and 72 women; mean age, 28 years; range, 22-51 years). All patients were stratified for histologic damage according to Marsh classification, and in all of them an anti tTG evaluation was performed. RESULTS: Marsh I lesions were present in 13 patients (10.92%), Marsh II in 24 anti-tTG (20.16%), Marsh IIIa in 27 anti-tTG (22.68%), Marsh IIIb in 31 anti-tTG (26.05%) and Marsh IIIc in 24 anti-tTG (20.16%). Anti-tTG positivity was ranging from 1 of 13 anti-tTG (7.69%) in Marsh I lesions to 23 of 24 anti-tTG (95.83%) in Marsh IIIc lesions respectively ( P< 0.005), while mean serum value of anti-tTG ranged from 3.61 (range, 0.7-9.2) UA/mL in Marsh I lesions to 7.3 (range, 1-25.1), 18.5 (range, 1.8-56.2), 36 (range, 3.7-83.5) and 74.95 (range, 6.5-257) UA/mL in Marsh II, IIIb and IIIc lesions respectively (P < 0.005). CONCLUSIONS: Our study showed that anti-tTG prevalence and their mean serum value was higher in celiacs with severe enteropathy (Marsh IIIb-c lesions) than in those showing slight enteropathy (Marsh I-IIIa). So, serologic tests without histologic evaluation may underestimate the real prevalence of CD and there is the risk of delaying the diagnosis of CD in patients who run an increased risk of deficiencies, non malignant conditions and malignancy. PMID- 12590233 TI - Number of responses on a review of systems questionnaire predicts the diagnosis of functional gastrointestinal disorders. AB - GOALS: We evaluated whether the number of positive responses on a review of systems questionnaire predicted the diagnosis of functional gastrointestinal (GI) disorders. STUDY: New patients to the University of Michigan GI clinics fill out a standardized questionnaire that includes a comprehensive review of systems which lists 87 items under 11 headings. We determined the number of items circled on the review of systems from the first 100 new patients seen in GI clinic in the year 2000. After a mean of 14 months follow-up, the final diagnoses in these patients were compared with the number of items circled on the review of systems. RESULTS: Completed questionnaires were available for review in 82/100 patients. Twenty-six patients were diagnosed with a functional GI disorder, 53 patients had a nonfunctional diagnosis, and 3 patients had no diagnosis determined. The mean number of items circled on the review of systems was 15.9 (SD = 11.2) for the functional GI group, compared with 5.9 (SD = 6.9) for the nonfunctional GI diagnosis group ( < 0.005). When the symptom headings were analyzed individually, the differences in general, gastroenterological, cardiovascular, urinary, neurological, eye, ear, nose and throat, and psychiatric symptoms were statistically significant. CONCLUSIONS: Patients with functional GI disorders have a significantly greater number of GI and non-GI symptoms on review of systems. If seven positive symptoms is used as a threshold, the standardized review of systems questionnaire had a sensitivity of 92% and specificity of 80% in detecting patients with functional GI disorders. PMID- 12590234 TI - First-line chemotherapy with fluorouracil and folinic acid for advanced colorectal cancer in elderly patients: a phase II study. AB - BACKGROUND: Colorectal cancer is one of the most common cancers in the elderly. Information on tolerability and efficacy of 5-Fluorouracil-based chemotherapy in such patients is limited. Primary aim of the study was to describe tolerability and activity of chemotherapy with the "de Gramont" schedule (FU bolus [400 mg/m ] + FU continuous infusion [600 mg/m ] + folinic acid [100 mg/m ] on days 1 and 2, every 2 weeks), in patients with advanced colorectal cancer aged 70 or older. PATIENTS AND METHODS: Patients aged 70 or more, with stage IV colorectal cancer, ECOG performance status not worse than 2. RESULTS: Thirty-four patients were treated at two participating centers. Seven (20.6%, 95% exact CI = 8.7-37.9) had an objective response, complete in 3 and partial in 4 patients. Five cases of unacceptable toxicity were registered (2 cardiac, 1 each for liver, anemia and diarrhea). Fitting the statistical model to the observed data indicated that the treatment was sufficiently active and tolerated. CONCLUSIONS: The de Gramont scheme is active and tolerated in elderly patients with advanced colorectal cancer. PMID- 12590235 TI - Hepatotoxicity associated with 6-thioguanine therapy for Crohn's disease. AB - 6-Thioguanine (6-TG) is a thiopurine analogue that is closely related to 6 mercaptopurine (6-MP) and azathioprine (AZA). These agents have potent cytotoxic and immunomodulatory effects and are useful in the treatment of a variety of conditions, including inflammatory bowel disease. Both 6-MP and AZA are widely used and are known to cause hepatotoxicity in a proportion of patients. 6 Thioguanine is being increasingly used in the treatment of inflammatory bowel disease but has not been reported to cause liver injury in this context. We describe a case of significant elevation of serum transaminases in a patient treated with 6-TG for a flare of Crohn's disease. We believe the temporal association of the abnormal liver enzymes in this patient, in the absence of other offending agents, argues strongly in favor of 6-TG as a cause of liver enzyme abnormalities. This case highlights the need to monitor liver enzymes in patients treated with 6-TG and identifies the need for additional research focused on the mechanism of thiopurine-induced hepatic injury. PMID- 12590236 TI - Infliximab for severe recurrent Crohn's disease presenting with massive gastrointestinal hemorrhage. AB - Severe gastrointestinal hemorrhage is an uncommon complication of Crohn's disease. Most bleeding episodes originate from colonic ulcers or ulcerated areas. The management of severe gastrointestinal bleeding in Crohn's disease is a therapeutic challenge. Several approaches including surgical resection, specific medical therapy of Crohn's disease, endoscopic treatment, or angiographic intervention have been attempted, but recurrence of bleeding is high. Monoclonal anti TNFalpha antibodies (infliximab) can induce relatively rapid mucosal healing. We report two cases of severe recurrent Crohn's disease presenting with massive lower gastrointestinal bleeding in which infliximab induced rapid mucosal healing and prevented recurrent bleeding. PMID- 12590237 TI - Hepatitis C and alcohol: interactions, outcomes, and implications. AB - BACKGROUND: Alcoholism and chronic hepatitis C (HCV) infection are common causes of liver disease worldwide. Hepatitis C virus and alcohol use frequently coexist, and together lead to more rapid progression of liver disease. GOALS: To critically review the literature pertaining to the combined effects of alcohol and HCV, focusing primarily on how alcohol influences the natural history, pathogenesis, and treatment of HCV liver disease. STUDY: A thorough review of the English literature was conducted, using a MEDLINE-based computerized literature search and review of cited references. RESULTS: Hepatitis C virus is prevalent in unselected alcoholic populations (14-36%) and in alcoholic individuals with liver disease (< or =51%). Hepatitis C virus-infected individuals who drink alcohol in excess have more severe histologic injury, more rapid disease progression, and a higher frequency of cirrhosis and hepatocellular carcinoma. Alcohol use also appears to decrease response rates to interferon therapy. The mechanisms of interaction between alcohol and HCV are not fully understood, but they likely include the effects of alcohol on the host immune system and the virus and on other factors possibly related to HCV liver disease and hepatic carcinogenesis. CONCLUSIONS: Alcohol use and HCV infection frequently coexist. Although there is ample evidence that alcohol use adversely affects the natural history of HCV liver disease, how the two interact is not well understood. Patients with chronic HCV should be encouraged to avoid alcohol; however, the threshold above which alcohol results in accelerated liver disease remains to be determined. PMID- 12590238 TI - Balthazar computed tomography severity index is superior to Ranson criteria and APACHE II and III scoring systems in predicting acute pancreatitis outcome. AB - BACKGROUND AND GOALS: Acute pancreatitis runs an unpredictable course. We prospectively analyzed the prognostic usefulness of four different scoring systems in separately assessing three variables; acute pancreatitis severity, development of organ failure and pancreatic necrosis. STUDY: 78 patients with acute pancreatitis were studied prospectively. Data pertinent to scoring systems were recorded 24 hours (APACHE II and III scores), 48 hours (Ranson score) and 72 hours (Balthazar computed tomography severity index) after admission. Statistical analysis was performed by using receiver operating characteristic curves and by comparing likelihood ratios of positive test (LRPT) for all three outcome variables. RESULTS: 44 patients were classified as mild and 34 as severe pancreatitis. When we compared LRPT, only that for the Balthazar score (11.2157) was able to generate large and conclusive changes from pretest to post-test probability in acute pancreatitis severity prediction. LRPT were 2.4157 for Ranson, 4.0980 for APACHE II and 3.6670 for APACHE III score. The APACHE II and III scores and Ranson criteria performed slightly better than the Balthazar score in predicting organ failure (LRPT: 4.0667, 3.2892, 3.0362 and 1.7941 respectively), while when predicting pancreatic necrosis the APACHE II and III performed slightly better than the Ranson score (LRPT: 2.0769, 2.7500 and 1.7813 respectively). CONCLUSIONS: In all outcome measures the APACHE scores generate small and of similar extent changes in probability. The Balthazar score is superior to other scoring systems in predicting acute pancreatitis severity and pancreatic necrosis. However the Ranson and APACHE scores perform slightly better with respect to organ failure prediction. PMID- 12590239 TI - Optimal management of the branch duct type intraductal papillary mucinous neoplasms of the pancreas. AB - BACKGROUND: To determine the optimal management of the intraductal papillary mucinous neoplasms (IPMNs) according to the morphologic type based on distinguishing between benign and malignant diseases. BACKGROUNDS: IPMNs are increasingly recognized clinicopathologic entity. Extended pancreatic resection with radical lymph node dissection has been recommended for treatment. STUDY: A retrospective clinicopathologic study was carried out of the 57 cases with IPMNs who were treated between 1985 and 2001. Forty-three patients with IPMNs underwent resection, and 14 patients with small IPMNs were observed without resection. RESULTS: Among the 43 resected IPMNs, 25 were benign and 18 were malignant. Malignant tumors were significantly greater in diameter than benign tumors (52.9 vs. 30.2 mm, P< 0.05). All main duct type tumors with mural nodules were malignant. All branch duct type tumors less than 30 mm in diameter and without mural nodules were benign. Twelve branch duct type IPMNs size less than 30 mm were not resected and have not progressed. CONCLUSION: These results suggest that the branch duct type IPMNs less than 30 mm and without mural nodules is benign and might be treatable with limited resection or careful observation. PMID- 12590240 TI - Probucol in the treatment of nonalcoholic steatohepatitis: an open-labeled study. AB - GOALS: To evaluate the effects of probucol, an agent with strong antioxidant properties, in reversing biochemical changes in nonalcoholic steatohepatitis (NASH). BACKGROUND: There is currently no well-established medical treatment of NASH. It is believed that oxidative stress plays a major role in hepatic damage in these patients. STUDY: Cases of biopsy-proven NASH referring to a referral center in Tehran during a 12-month period were included in the study. Viral, autoimmune and other hepatic diseases were excluded. Alcohol ingestion was excluded by repeated questioning of the patient and at least two family members. Patients were given 500mg of probucol daily for 6 months. Serum levels of liver enzymes, the serum lipid profile, and weight was recorded monthly. RESULTS: A total of 17 patients completed the study. The mean age was 37.2 years, 13 patients were male and 4 female. The mean pretreatment value of ALT and AST was 93.5 and 80.4 U/L, and the mean posttreatment value was 41.8 and 35.9 U/L respectively ( = 0.001 and 0.006). CONCLUSION: Probucol, even in the low dose of 500 mg/d, appears to be significantly effective in decreasing the ALT and AST levels in patients with NASH. PMID- 12590241 TI - Nuclear localization of hepatitis B core antigen and its relations to liver injury, hepatocyte proliferation, and viral load. AB - GOALS: The aim of this study was to determine the factor(s) independently affecting the HBcAg expression pattern in HBV-infected livers. BACKGROUND: Subcellular localization of HBcAg have been found to be related to the activity of liver disease, hepatocyte proliferation rate and the level of HBV replication. STUDY A total of 98 patients with biopsy proven chronic hepatitis B were included. HBcAg and proliferative cell nuclear antigen (PCNA) were immunohistochemically detected. HBcAg expression and its relationship with histologic activity index, ALT levels, PCNA score and HBV DNA levels were investigated. RESULTS: Forty-three (44%) patients were positive for HBcAg staining, with most of them (37 patients) having nuclear localization. Forty seven percent of patients with detectable HBV DNA had nuclear staining while none of the HBV DNA negative patients had nuclear staining ( < 0.0005). Patients with positive nuclear staining had lower HAI (<8) and a lower PCNA score (<353/1,000 cell) than those with negative staining (62% vs. 39% and 90% vs. 66%, respectively, P< 0.05). In multivariate analysis, both high HBV DNA level and low HAI ( P< 0.0005 and P< 0.05, respectively) but not PCNA score, were independently associated with nuclear staining of HBcAg. CONCLUSIONS: Our results suggest that viral load and the severity of liver damage but not the rate of hepatocyte proliferation independently affects the HBcAg expression pattern. PMID- 12590242 TI - Candesartan cilexetil-induced severe hepatotoxicity. AB - We report a case of severe hepatotoxicity associated with ductopenia in a 61-year old woman. The patient developed jaundice, vomiting, and abdominal pain a few weeks after the beginning of treatment with candesartan cilexetil, 16 mg/d, for essential hypertension. Liver biopsy showed parenchymal bilirubinostasis with portal cholangitis and ductopenia. The drug was immediately withdrawn, and in the following weeks, aminotransferases and serum bilirubin returned to normal levels. The clinical-histologic data and the exclusion of known causes of liver disease led us to make a diagnosis of drug-induced cholestasis. To our knowledge, this is the first description of severe hepatotoxicity associated with ductopenia caused by an adverse reaction to candesartan cilexetil. PMID- 12590243 TI - Hepatitis B virus reactivation after kidney transplantation and new onset lymphoma. AB - A case of hepatitis B virus (HBV) reactivation after kidney transplantation is reported. The presence of antibodies against hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) before transplantation indicated previous infection with HBV. Eight years after transplantation, a diffuse, large B-cell lymphoma occurred, and the patient was positive for HBsAg and hepatitis B e antigen, in association with normal activity of serum aminotransferases. Postmortem liver histology did not show any sign of portal tract or lobular inflammation despite the presence at immunostaining of extensive intranuclear and cytoplasmic positivity for HBcAg, indicating active viral replication. Natural immunity to HBV may not protect against reactivation in patients with a suppressed immune system. In this setting, periodic follow-up of HBV serology in patients at highest risk for HBV reactivation to allow for early diagnosis and prompt treatment with lamivudine is highly recommended. PMID- 12590244 TI - Prediction of fatty liver: comparison of body mass index, visceral fat, and subcutaneous fat. PMID- 12590245 TI - A young girl with recurrent severe abdominal pain after appendicectomy. PMID- 12590246 TI - Erythematous maculopapular eruption due to ribavirin administration in a patient with chronic hepatitis C. PMID- 12590247 TI - The "corrected" half emptying time is a valueless parameter in the 13C-octanoic acid breath test: misuse of the regression analysis. PMID- 12590248 TI - Association of Clostridium difficile infection with collagenous colitis. PMID- 12590251 TI - New dimensions for medical mycology in the 21st century. PMID- 12590252 TI - The changing clinical presentation of fungal infections. PMID- 12590253 TI - [Intraspecies diversity of Trichosporon asahii as the causative agent of opportunistic fungal infection and summer-type hypersensitivity pneumonitis]. AB - Trichosporon asahii is the major causative agent of the opportunistic infection trichosporonosis, and also causes summer-type hypersensitivity pneumonitis (SHP). Random amplification of polymorphic DNA analysis was used to determine the intraspecies diversity of T. asahii isolates from clinical specimens,the houses of SHP patients, and environmental material. Clinical isolates formed a cluster, characterized by a 90% matching coefficient, but they did not cluster with strains isolated from SHP patients houses or environmental sources. The biochemical characteristics of T. asahii isolates from the three sources were compared, and a phenogram was constructed. One of the two clusters included most of the clinical isolates and strains isolated from the houses, and the other cluster included most of the environmental isolates. There was a remarkable difference in the abilities of the strains belonging to these clusters to utilize several compounds. These results suggest that the genetic diversity and biochemical characteristics of T. asahii are related to the source of the isolates. In addition, based on the IGS1 sequence, which is located between the 26S and 5S rRNA genes, we identified five genotypes of T. asahii, which is a major causative agent of deep-seated trichosporonosis. Of the five genotypes, three were isolates that originated in Japan, whereas two were American isolates. IGS sequence analysis shows great potential as a new epidemiological tool. PMID- 12590254 TI - [Molecular biological analysis of Tinea faciei caused by a clinically anomalous strain of Trichophyton mentagrophytes]. AB - We report a case of Trichophyton mentagrophytes infection. A 27-year-old female dairy worker visited our hospital in August 2001 for evaluation of an erythematous lesion near her right eye. The patient had been treated with antibiotics and an anti-allergic drug at another clinic with no effect on the lesion. KOH preparation of the scale revealed mycelia and T. mentagrophytes was isolated from the culture. The isolated pathogen was identified as T. mentagrophytes animal type 1 by the classification system based on the cluster analysis of nuclear ribosomal internal transcribed spacer 1 (ITS1) DNA sequences. The oral administration of terbinafine at 125 mg/day and antifungal cream was effective. T. mentagrophytes is one of the most common dermatophytes isolated from humans and animals. We should be aware that animals may carry this kind of fungal pathogen. PMID- 12590255 TI - Direct invasion of bones by highly pathogenic fungi in an in vitro model and its ecological significance. AB - Animal bones after being devitalized at death are strongly resistant to wear and tear and remain in the soil or environment much longer than other organic components from dead animals. Yet over the course of time they seem to disappear and thus our ecological surroundings are not cluttered with bone remnants. Mechanical factors creating compression or friction and chemical factors like pH of the soil and surroundings must together have provided concerted degrading effects. Microorganisms in the soil also help in this process by utilizing the organic components of devitalized bones. Certain highly pathogenic fungi that have been collected from soil from time to time and many other environmental fungi may take part in the degrading of the bone remnants. In this study, several strains from the highly pathogenic dimorphic fungi Coccidioides immitis, Blastomyces dermatitidis, Histoplasma spp., Paracoccidioides brasiliensis and also some strains of dematiaceous fungi (Exophiala spp. and Foncecaea pedrosoi) were inoculated to dissected and devitalized murine long bones that had been placed on solidified water agar plates to see if they would survive, grow and invade the bones. After being kept for 12 weeks at 25 degrees C all the parts of the histological sections of these bones showed invasion by most of the strains used in this study, although the cortical component of the bony architecture seemed to be comparatively resistant to invasion. Their ability to grow and sporulate in the aforementioned nutrient-limiting condition hinted at a possible role of these fungi in the degradation of devitalized bones. PMID- 12590256 TI - [A case of Tinea capitis caused by Trichophyton tonsurans]. AB - A 10-year-old Peruvian girl, living in Japan since 1996, visited our hospital in August 2000 complaining of alopecia which had been present on her scalp for one year. The bald areas appeared as multiple small, scattered, angular patches with indistinct margins. Follicular pustules, erythemic nodules and lymphadenopathy were also seen. In the culture of the affected hair, a tan surface with wiry undulations grew on Sabouraud's media. The colony reverse had reddish-brown central pigmentation. Slide cultured fungi produced great numbers of round and short club-shaped microconidia, hyphae and intercalary chlamydospores. These fungi showed the following characteristics: positive urease test, no pigment production on cornmeal agar and positive thiamine dependency. The restriction fragment length polymorphism pattern and the nucleotide sequences of ribosomal DNA internal transcribed spacer region of the causative fungus was compatible with Trichophyton tonsurans. Daily administration of 125 mg of terbinafine resulted in a satisfactory response and the lesion healed almost completely. PMID- 12590257 TI - The epidemiology and mating behavior of Arthroderma benhamiae var. erinacei in household four-toed hedgehogs (Atelerix albiventris) in Japan. AB - An epidemiological survey of Trichophyton mentagrophytes var. erinacei in the household hedgehog and other rodents was made between January 17, 2002 and February 28, 2002 in Japan. Quills and hairs were collected from sources identified via the internet. The fungus was isolated only from the quills of four toed hedgehogs (7/18; 39%) from Kanto to Kyushu regions. Isolates were examined morphologically, physiologically and genetically, and identified as T. mentagrophytes var. erinacei anamorph. The isolates were also genetically compared with European hedgehog (Erinaceus europeus)-borne T. mentagrophytes var. erinacei and Kenyan hedgehog (Aterelix albiventris)-borne Arthroderma benhamiae, and their genotypes of the ITS1-5.8S-ITS2 rDNA were all identical. The isolates were crossed with A. benhamiae Americano-European race and African race, A. vanbreuseghemii and A. simii, with the result that they mated only with African race (+) or (-). Mating types of the isolates were (+) in 6 isolates and (-) in one. An intra-isolate mating between one of the 6 plus isolates and the minus one formed abundant mature gymnothesia, the mating type ratio of the F1 progeny was approximately 1:1, and the sib crossings of F1 progeny produced abundant fertile gymnothesia. The present study revealed that the intra-Japanese hedgehog-borne isolate crossing showed complete fertility and that the sexual degeneration pointed out by Takashio (Mycologia 71: 968-976, 1979) did not exist. Two pairs of mating, (+) and (-) mating types of Japanese isolates with (-) and (+) tester strains of A. benhamiae African race formed less gymnothesia, mating type ratios were unbalanced, and sib crossings of F1 progeny produced small gymnothesia containing a low number of asci, pseudogymothesia, or none, respectively. These results show that A. benhamiae var. erinacei, the teleomorph of T. mentagrophytes var. erinacei, belongs to a different mating group (e.g. hedgehog race) than the Americano-European and African races in A. benhamiae. PMID- 12590258 TI - Disruption of Cnp1 uncouples oligodendroglial functions in axonal support and myelination. AB - Myelination of axons by oligodendrocytes enables rapid impulse propagation in the central nervous system. But long-term interactions between axons and their myelin sheaths are poorly understood. Here we show that Cnp1, which encodes 2',3'-cyclic nucleotide phosphodiesterase in oligodendrocytes, is essential for axonal survival but not for myelin assembly. In the absence of glial cyclic nucleotide phosphodiesterase, mice developed axonal swellings and neurodegeneration throughout the brain, leading to hydrocephalus and premature death. But, in contrast to previously studied myelin mutants, the ultrastructure, periodicity and physical stability of myelin were not altered in these mice. Genetically, the chief function of glia in supporting axonal integrity can thus be completely uncoupled from its function in maintaining compact myelin. Oligodendrocyte dysfunction, such as that in multiple sclerosis lesions, may suffice to cause secondary axonal loss. PMID- 12590259 TI - Impaired response to interferon-alpha/beta and lethal viral disease in human STAT1 deficiency. AB - The receptors for interferon-alpha/beta (IFN-alpha/beta) and IFN-gamma activate components of the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway, leading to the formation of at least two transcription factor complexes. STAT1 interacts with STAT2 and p48/IRF-9 to form the transcription factor IFN-stimulated gene factor 3 (ISGF3). STAT1 dimers form gamma-activated factor (GAF). ISGF3 is induced mainly by IFN-alpha/beta, and GAF by IFN-gamma, although both factors can be activated by both types of IFN. Individuals with mutations in either chain of the IFN-gamma receptor (IFN-gammaR) are susceptible to infection with mycobacteria. A heterozygous STAT1 mutation that impairs GAF but not ISGF3 activation has been found in other individuals with mycobacterial disease. No individuals with deleterious mutations in the IFN alpha/beta signaling pathway have been described. We report here two unrelated infants homozygous with respect to mutated STAT1 alleles. Neither IFN-alpha/beta nor IFN-gamma activated STAT1-containing transcription factors. Like individuals with IFN-gammaR deficiency, both infants suffered from mycobacterial disease, but unlike individuals with IFN-gammaR deficiency, both died of viral disease. Viral multiplication was not inhibited by recombinant IFN-alpha/beta in cell lines from the two individuals. Inherited impairment of the STAT1-dependent response to human IFN-alpha/beta thus results in susceptibility to viral disease. PMID- 12590260 TI - Amnionless, essential for mouse gastrulation, is mutated in recessive hereditary megaloblastic anemia. AB - The amnionless gene, Amn, on mouse chromosome 12 encodes a type I transmembrane protein that is expressed in the extraembryonic visceral layer during gastrulation. Mice homozygous with respect to the amn mutation generated by a transgene insertion have no amnion. The embryos are severely compromised, surviving to the tenth day of gestation but seem to lack the mesodermal layers that normally produce the trunk. The Amn protein has one transmembrane domain separating a larger, N-terminal extracellular region and a smaller, C-terminal cytoplasmic region. The extracellular region harbors a cysteine-rich domain resembling those occurring in Chordin, found in Xenopus laevis embryos, and Sog, found in Drosophila melanogaster. As these cysteine-rich domains bind bone morphogenetic proteins (Bmps), it has been speculated that the cysteine-rich domain in Amn also binds Bmps. We show that homozygous mutations affecting exons 1-4 of human AMN lead to selective malabsorption of vitamin B12 (a phenotype associated with megaloblastic anemia 1, MGA1; OMIM 261100; refs. 5,6) in otherwise normal individuals, suggesting that the 5' end of AMN is dispensable for embryonic development but necessary for absorption of vitamin B12. When the 5' end of AMN is truncated by mutations, translation is initiated from alternative downstream start codons. PMID- 12590261 TI - Notch1 functions as a tumor suppressor in mouse skin. AB - Notch proteins are important in binary cell-fate decisions and inhibiting differentiation in many developmental systems, and aberrant Notch signaling is associated with tumorigenesis. The role of Notch signaling in mammalian skin is less well characterized and is mainly based on in vitro studies, which suggest that Notch signaling induces differentiation in mammalian skin. Conventional gene targeting is not applicable to establishing the role of Notch receptors or ligands in the skin because Notch1-/- embryos die during gestation. Therefore, we used a tissue-specific inducible gene-targeting approach to study the physiological role of the Notch1 receptor in the mouse epidermis and the corneal epithelium of adult mice. Unexpectedly, ablation of Notch1 results in epidermal and corneal hyperplasia followed by the development of skin tumors and facilitated chemical-induced skin carcinogenesis. Notch1 deficiency in skin and in primary keratinocytes results in increased and sustained expression of Gli2, causing the development of basal-cell carcinoma-like tumors. Furthermore, Notch1 inactivation in the epidermis results in derepressed beta-catenin signaling in cells that should normally undergo differentiation. Enhanced beta-catenin signaling can be reversed by re-introduction of a dominant active form of the Notch1 receptor. This leads to a reduction in the signaling-competent pool of beta-catenin, indicating that Notch1 can inhibit beta-catenin-mediated signaling. Our results indicate that Notch1 functions as a tumor-suppressor gene in mammalian skin. PMID- 12590262 TI - Chromosome-wide distribution of haplotype blocks and the role of recombination hot spots. AB - Recent studies of human populations suggest that the genome consists of chromosome segments that are ancestrally conserved ('haplotype blocks'; refs. 1 3) and have discrete boundaries defined by recombination hot spots. Using publicly available genetic markers, we have constructed a first-generation haplotype map of chromosome 19. As expected for this marker density, approximately one-third of the chromosome is encompassed within haplotype blocks. Evolutionary modeling of the data indicates that recombination hot spots are not required to explain most of the observed blocks, providing that marker ascertainment and the observed marker spacing are considered. In contrast, several long blocks are inconsistent with our evolutionary models, and different mechanisms could explain their origins. PMID- 12590263 TI - The study of macromolecular complexes by quantitative proteomics. AB - We describe a generic strategy for determining the specific composition, changes in the composition, and changes in the abundance of protein complexes. It is based on the use of isotope-coded affinity tag (ICAT) reagents and mass spectrometry to compare the relative abundances of tryptic peptides derived from suitable pairs of purified or partially purified protein complexes. In a first application, the genuine protein components of a large RNA polymerase II (Pol II) preinitiation complex (PIC) were distinguished from a background of co-purifying proteins by comparing the relative abundances of peptides derived from a control sample and the specific complex that was purified from nuclear extracts by a single-step promoter DNA affinity procedure. In a second application, peptides derived from immunopurified STE12 protein complexes isolated from yeast cells in different states were used to detect quantitative changes in the abundance of the complexes, and to detect dynamic changes in the composition of the samples. The use of quantitative mass spectrometry to guide identification of specific complex components in partially purified samples, and to detect quantitative changes in the abundance and composition of protein complexes, provides the researcher with powerful new tools for the comprehensive analysis of macromolecular complexes. PMID- 12590266 TI - Genetics and heart disease. AB - Production of genome sequence has recently skyrocketed with many advances in the understanding and etiology of certain diseases. Researchers have localized a region of the human genome that plays a role in determining a persons susceptibility to myocardial infarction. A new apolipoprotein gene that influences triglyceride levels in humans is also described. A recent study from Finland showed that certain families are likely to carry a genetic form of insulin resistance syndrome that predisposes them to accelerated atherosclerosis. Researchers identified 3 mutations in the gene producing a protein called metavinculin, which appears to be linked to abnormalities in cellular structures and function in patients with dilated cardiomyopathy. Gene therapy has emerged as a genuine therapeutic option with the potential to alter the manner in which cardiologists manage the 2 most common cardiac disorders--coronary artery disease and congestive heart failure. Along with angiogenesis and gene therapy, cell transplantation is one of the newest treatment modalities proposed to improve the outcome of patients with cardiac failure. Two major advances in stem cell therapy for cardiovascular disease were published recently. They demonstrate how bone marrow stem cells can regenerate myocardium in the infarct area of a mouse heart. A German Cardiologist has for the first time successfully transplanted a patients own stem cells in an infracted area in the heart. This review summarizes the current knowledge of the genetic associations with cardiac diseases. PMID- 12590264 TI - A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. AB - RNA interference (RNAi) has recently emerged as a specific and efficient method to silence gene expression in mammalian cells either by transfection of short interfering RNAs (siRNAs; ref. 1) or, more recently, by transcription of short hairpin RNAs (shRNAs) from expression vectors and retroviruses. But the resistance of important cell types to transduction by these approaches, both in vitro and in vivo, has limited the use of RNAi. Here we describe a lentiviral system for delivery of shRNAs into cycling and non-cycling mammalian cells, stem cells, zygotes and their differentiated progeny. We show that lentivirus delivered shRNAs are capable of specific, highly stable and functional silencing of gene expression in a variety of cell types and also in transgenic mice. Our lentiviral vectors should permit rapid and efficient analysis of gene function in primary human and animal cells and tissues and generation of animals that show reduced expression of specific genes. They may also provide new approaches for gene therapy. PMID- 12590267 TI - Anti-spasticity medications. AB - Spasticity is common in patients with a variety of central nervous system disorders. It can lead to significant disability or cause complications that may result in severe morbidity. In such patients, treatment of spasticity is warranted. Several oral and parenteral medications are available for use in the treatment of spasticity. This article reviews the pharmacological properties and therapeutic effectiveness of these medications to provide a practical objective guide for physicians who may be involved in the management of spasticity. PMID- 12590268 TI - Effects of Ramadan fasting on serum lipid profiles on 2 hyperlipidemic groups with or without diet pattern. AB - OBJECTIVE: The effects of Ramadan fasting, with low fat and low calorie diet, on blood lipid and lipoprotein levels were studied. Results revealed reduction of plasma lipid levels and anthropometric parameters in the hyperlipidemic cases. To find out whether such reductions were due to nutritional diet or Ramadan fasting, we conducted a study to evaluate effects of Ramadan fasting on 2 separate hyperlipidemic groups with or without nutritional diet regimen. METHODS: This study was carried out at Madani Heart Hospital, Tabriz, Iran, during the year 1998. Thirty-eight hyperlipidemic healthy men voluntarily enrolled into 2 groups, group I, 22 men on low fat and low calorie diet and group II, 16 men without any special diet interference. The blood lipid profile tests were measured 4 times (3 weeks before, first week, last week and one month after Ramadan). To evaluate nutritional composition, 12 times in non-successive days, 24 hour nutrition recalls were obtained from all individuals during the study. RESULTS: Analysis of data revealed that only triglyceride in both groups reduced in the beginning of Ramadan compared to 3 weeks before. During Ramadan, with a reduction of 300 Kcal/day in comparison to before Ramadan, no changes were seen concerning anthropometric parameters and serum lipids levels. CONCLUSION: It seems that the effect of Ramadan fasting on serum lipid levels may be closely related to the nutritional diet. For reduction of plasma lipid levels, it would be necessary to omit at least one term meal or reduce energy by 500 Kcal or more per day. PMID- 12590269 TI - Obesity among Saudi male adolescents in Riyadh, Saudi Arabia. AB - OBJECTIVE: The purpose of the study was to determine the prevalence of overweight and obesity and its correlates among Saudi male adolescents in Riyadh. As well as evaluating their knowledge, attitude and practice towards obesity. METHODS: A cross-sectional study conducted in intermediate and secondary schools in Riyadh, Saudi Arabia was carried out during a 5-month period, September 2001-January 2002. A sample of 894 Saudi male adolescents (age 12-20 years) was selected through the multi-stage sampling technique. Socio-demographic characteristics; dietary and activity history; obesity-related knowledge and behavior; and family and past medical history data were obtained by a self-administered questionnaire. Anthropometric measurements of weight and height were performed. Body mass index (BMI) was calculated, and adolescents with a BMI age-specific percentile of > or = 85th-< 95th were considered overweight and > or = 95th were considered obese. RESULTS: The prevalence of overweight was 13.8% and obesity was 20.5%. Family history (odds ratio, 2.49; 95% confidence interval, 1.72-3.61) and lack of physical activity (odds ratio, 1.63; 95% confidence interval, 1.01-2.62) were associated with adolescent obesity. Twenty percent of overweight participants did not think they were overweight. CONCLUSION: Obesity constitutes an important public health problem among male adolescents in Riyadh. A national prevention program with involvement of schools is recommended to avoid obesity-related morbidity in adulthood. PMID- 12590270 TI - Luteal phase support in ovarian induction cycles using human chorionic gonadotropin or oral progestagens. AB - OBJECTIVE: To determine the efficacy of luteal phase support with human chorionic gonadotropin (hCG) or oral progesterone during human menopausal gonadotropin (hMG) ovulation induction. METHODS: Between September 1999 and March 2001, a total of 91 couples with infertility were recruited at Al-Hammadi Hospital, Riyadh, Kingdom of Saudi Arabia and Badeea Hospital, Jordan. In this prospective trial 46 couples were allocated to luteal phase support with hCG injections, while 45 couples were allocated to Duphaston (oral progestogens) as luteal support. RESULTS: In the group of hCG luteal support, 46 patients completed 46 cycles of hMG therapy, and 8 pregnancies (5 ongoing pregnancies) ensued, with a general total pregnancy rate of 17.4%. In the progesterone (Duphaston) luteal support group, 45 patients with the similar indications to the previous group were studied and 8 pregnancies (5 ongoing pregnancies) were reported with a general total pregnancy rate of 17.8%. Only one spontaneous abortion occurred among the patients in a cycle supported with supplemental hCG, while 2 abortions occurred in the Duphaston supported group. CONCLUSION: Despite theoretical reasons to use luteal phase support during hMG-stimulated cycles, our data showed no improvement in pregnancy rates from such treatment. PMID- 12590271 TI - Uterine rupture incidence, risk factors, and outcome. AB - OBJECTIVE: To determine the incidence of uterine rupture and to analyze risk factors and outcome in a local hospital in Makkah, Kingdom of Saudi Arabia, (KSA). METHODS: The hospital records of the Maternity and Children's Hospital (MCH), in Makkah, KSA from April 1999 to December 2000 were retrospectively reviewed to identify women with ruptured uterus. The relevant data relating to the clinical features, risk factors, operative procedures, and maternal and fetal outcomes were assessed. RESULTS: During the study period, there were 23245 deliveries and 23 women were diagnosed to have uterine rupture giving an incidence of one in 1011 deliveries. Fifteen (65.2%) occurred in women with previous cesarean scar and 8 (34.8%) women had no previous uterine surgery. In the 15 women with uterine rupture and previous cesarean section there was no maternal death. They were treated by repair of the uterus. Two women sustained bladder injury, and one subsequently developed vesico-vaginal fistula. In contrast, in the 8 women with no previous uterine surgery, one woman died, one woman developed renal failure, and there were 3 fetal losses. Four women needed total abdominal hysterectomy, and 4 women needed repair. Two women needed internal iliac ligation in addition to the hysterectomy. CONCLUSION: In our circumstances, uterine rupture is not rare and consequences can be life threatening. The outcome is worse in women with unscarred uterus. PMID- 12590272 TI - A scoring system as a tool for antenatal care audit in family medicine. AB - OBJECTIVE: To propose the antenatal assessment score (AAS) as a tool for auditing the process of antenatal care in family medicine, and to highlight its accessibility by applying it at 2 family health care centers. METHODS: A descriptive study of an audit process was conducted in 2 primary health care centers (non commissioned officers' [NCO] and officers' health centers [OC]) belonging to the Department of Family Medicine, Armed Forces Hospital, Riyadh, Kingdom of Saudi Arabia from February 2001 to June 2002. A systematic random sample of patients registered at the maternal registers of both centers was selected. A score system of 100 points was developed containing items regarding history taking, examination, investigations, treatment, referral, health education, number of visits and record keeping. The information was gathered retrospectively from patients' files at both centers. RESULTS: The total assessment score approached 67 out of 100 for NCO and 71 out of 100 for OC with a statistically significant difference (p<0.05). History taking in general was achieved to 77% in NCO and 86% in OC. Examination whether general (8% and 76%) or obstetric (67% and 72%) was achieved at a lesser level. All investigations were recorded equally in both centers (77%). Referrals, whether routine or emergency, were much less recorded 35-28%. The majority of antenatal records at both centers were partially completed (84.5% and 81%). Indications to any given medications (73% and 91%) and the total number of visits were higher among NCO records with statistically significant difference (p<0.05). CONCLUSION: Antenatal assessment score is an accessible tool for the audit process of antenatal care in family medicine. In depth analysis and interpretation of the results could be of high importance to total antenatal care. Nationwide use of this audit tool is recommended. PMID- 12590273 TI - Response to beta interferon 1b among Saudi patients with multiple sclerosis. AB - OBJECTIVE: To determine the efficacy and tolerability of subcutaneous beta interferon 1b (B1F1b) among Saudi patients with remitting-relapsing multiple sclerosis (R-R MS). METHODS: An open label study held at the Neurology Division of the Armed Forces Hospital, Riyadh from March 1997 until December 2001. Thirty two consecutive patients below the age of 50 years with clinically definite R-R MS according to Poser's Criteria and expanded disability status scale below 5.5 were enrolled in treatment with subcutaneous B1F1b 8 million IU 3 times a week. The primary outcome measures used were: reduction in annual relapses, proportion of relapse-free patients, and the mean time to the first relapse after treatment was started. The secondary outcome measures used were the time to progression in disability, tolerability and safety of the beta interferon. RESULTS: Only 28 patients were analyzed to assess the outcome measures, the other 4 patients dropped out and were followed-up. Twenty were women and 8 were men (female:male ratio of 2.5:1). There was a significant reduction in relapse-rate in all patients, 32.5% were relapse-free, while 37.5% showed reduction in the number of relapses. None of our patients showed progression of disability (P<0.0249). Mild adverse reactions were seen in 38.5%, influenza-like illness occurred in 53.6%, and injection-site reaction in 35.7%. CONCLUSION: Subcutaneous B1F1b is effective in patients with R-R MS, especially in reducing relapse rate, probable disability, and it is well tolerated. However, longer follow-up is necessary to evaluate the role of B1F1b in preventing disability. PMID- 12590274 TI - Prevalence and correlates of nocturnal enuresis in the United Arab Emirates. AB - OBJECTIVE: To determine the prevalence, socio-demographic correlates and associated psychopathology in children with enuresis. METHODS: A community survey of a stratified sample of 400 6-16 year old school children was carried out in the Al-Ain City, United Arab Emirates. Psychiatric disturbance, if any, was ascertained using the Rutter Parent Scale. RESULTS: Thirty-two out of the 400 children (8%) surveyed, reported wetting the bed at least once per week. More boys were affected than girls, and 90% of those with enuresis were aged 6-11 years. Presence of enuresis was found to be associated with psychosocial stress in the family; large family size; a positive family history, poorer scholastic performance and presence of psychiatric disturbance as indicated by the Rutter parent score. CONCLUSION: Our findings suggest that bed-wetting is a common problem among school children and that they need to be carefully screened for associated psychopathology. PMID- 12590275 TI - Cauterization of inferior nasal turbinate for idiopathic headache. AB - OBJECTIVE: Idiopathic headache is a stressful complaint, whose treatment is not well established. Our study is an attempt to examine the relationship between idiopathic headache and inferior turbinate cauterization as a surgical treatment, and to explain the central physiological effect of cauterization. METHODS: Thirty four patients, 15 females and 19 males complaining of idiopathic headache were treated by inferior turbinate cauterization. Cauterization of the inferior-medial aspect of the turbinate was carried out once under local anesthesia for few seconds. This study was performed between November 1993 and December 1996, a joint project of the Zarka Government Hospital, Zarka and Faculty of Medicine, University of Jordan, Amman, Jordan. RESULTS: More than 82% of the patients showed significant improvement in the sense of decreased headache. CONCLUSION: Cautery of the nasal turbinate could be a novel method for treatment of idiopathic headache. PMID- 12590276 TI - Hearing screening of neonates at risk. AB - OBJECTIVE: The aim of this study is to screen infants admitted to the Neonatal Intensive Care Unit (NICU) for hearing loss METHODS: One hundred and thirty newborns admitted to NICU were screened for hearing loss using otocaustic emission (OAE). Tympanometry was performed using Grason Stadler Incorporation (GSI) 37 and auditory brain response (ABR). RESULTS: Ninety-six of the infants had known risk factors such as prematurely, low birth weight, hyperbilirubinemia, asphyxia and congenital abnormalities. Out of 130 infants 80 passed the OAE test, 50 had to be rescreened, and 19 had ABR. Only 13 were found with moderate to severe hearing loss, 13.5% of at the risk infants. CONCLUSION: Hearing aids were provided from the age of 6 months, for 7 of the hearing impaired children. Parents of other children refused hearing aids and thought the child would talk when older, although they were full informed regading their children's hearing loss. Early identification and early intervention gives chances for infants to acquire speech and language. Parent's awareness will be stressed upon. PMID- 12590277 TI - Outcome of type-1 tympanoplasty. AB - OBJECTIVE: The outcome of 97 patients who underwent tympanoplasty type-1 surgery by different ear, nose and throat surgeons was evaluated. METHODS: This retrospective study was carried out from the records pertaining to all patients who had tympanoplasty type-1 at King Fahd Specialist Hospital, Buraidah, Al Qassim, Kingdom of Saudi Arabia, during the years April 1995 through to April 2000. Specific criterias were setup for selecting all the patients involved and excluding others. Thus 97 ears were selected in this study. The factors presumed to influence the outcome included age, sex, nationality, affected ear, middle ear status at surgery, perforation size and surgical approach and techniques chosen by different surgeons. These data was collected in a set out proformae and tables and analyzed and discussed. RESULTS: The patients age ranged from 11-45 years with a mean of 26.3, standard deviation 7.6 in all selected patients and also in the successful group. Very few children from age of 11-16 were involved. The male female ratio was 1:1.4 in the total selected group and 1:1.6 in the successful group. The success rate was 80% in males and 87.7% in females (p value 0.301). The results obtained showed no significant difference due to these factors except the permeatal approach (p<0.045) as compared to endaural approach. In this series the overall graft success rate was 84.5% in the total of 97 patients selected for the study. CONCLUSION: The overall success rate coincided with published literature from elsewhere. The factors presumed to influence the outcome of tympanoplasty type-1 surgery showed no significant difference, except the permeatal approach as compared to endaural approaches. PMID- 12590278 TI - Otolaryngologic manifestation of gastroesophageal reflux in children. AB - OBJECTIVE: This study was performed to assess the role of gastroesophageal reflux (GER) in varieties of otolaryngologic dysfunction. METHODS: This study was carried out over a one year period, between January 1995 and January 1996, at Sainte-Justine Hospital, Montreal, Canada. Thirty-seven charts were examined for different variables including, symptoms, signs, dysfunction, investigations, treatment and outcome. RESULTS: In children with otolaryngologic manifestation of GER, apnea and regurgitation were common presenting symptoms. Gastroesophageal reflux was also found to be more commonly associated with ear infection, adenoid hypertrophy, bronchial asthma and recurrent pneumonia. CONCLUSION: Gastroesophageal reflux has complicated manifestations in otolaryngology that are difficult to prove. High index of suspicion is recommended in order to diagnose GER as a cause of some otolaryngologic dysfunction. PMID- 12590279 TI - The pattern of esophageal manometry in progressive systemic sclerosis. AB - OBJECTIVE: The purpose of this paper is to study the esophageal motility pattern and the frequency of acid reflux in patients diagnosed to have progressive systemic sclerosis and compare the results to that of normal controls. METHODS: All consecutive patients diagnosed to have progressive systemic sclerosis between 1417-1419 (Hijra year) at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia were included (Group I). History of heartburn, dysphagia and regurgitation was reviewed. Drugs that might suppress acid or alter motility were discontinued 2 weeks before inclusion. The results were compared to that of 21 symptomatic controls of similar age and sexes that were seen at the same period (Group II). Esophageal manometry and upper gastrointestinal endoscopy were performed in all patients. Ambulatory 24 hour-pH monitoring was carried out in 6 patients of group 1 and 20 patients of group II. RESULTS: Thirteen progressive systemic sclerosis patients (12 females) mean age was 38.7 years and 21 (19 females) mean age was 34.8 years were included. The symptom scores, lower esophageal sphincter pressure, esophageal contractions amplitude were significantly worse in patients compared to control, dysphagia was mostly due to aperistalsis. All progressive systemic sclerosis patients showed the typical esophageal manometry pattern of lower esophageal sphincter pressure and diminished amplitude with aperistalsis. Gastroesophageal reflux was detected in 83% of patients with progressive systemic sclerosis. Moreover, all 24-hour pH monitoring variables were significantly worse in group I. CONCLUSION: Patients with progressive systemic sclerosis usually present with heartburn, dysphagia and regurgitation. Esophageal manometry typically shows lower pressure and aperistalsis. Gastroesophageal reflux is frequent. PMID- 12590280 TI - Profile of acute pancreatitis in Jizan, Saudi Arabia. AB - OBJECTIVE: The epidemiology of acute pancreatitis in the Kingdom of Saudi Arabia (KSA) may be different from the West. The present study describes the profile of acute pancreatitis in Jizan, KSA. METHODS: Patients diagnosed as acute pancreatitis in the King Fahd Central Hospital (KFCH), Jizan, KSA over a period of 12 years (1411-1422 Hijra year) were retrospectively analyzed with particular reference to the clinical features, disease severity, and result on management. RESULTS: There were 71 episodes in 62 patients (26 males and 36 females; 53 Saudis, whose ages ranged from 13-82 years (mean age: 42.6 years.). Of these patients, 5 had 2 episodes each one had 5 recurrent episodes. The etiology included gallstone disease in 26 (42%) cases; endoscopic retrograde cholangiopancreatography (ERCP) in 11 (18%) cases. No cause could be identified in 16 (26%). Patients. with ERCP - related pancreatitis were asymptomatic. In the symptomatic group (60 episodes), abdominal pain (100%), vomiting (78%) and fever (20%) were the most frequent features. Overall the clinical course was mild in 61 (86%) episodes and severe in 10 (14%). Ranson's scoring accurately identified 60 of the 61 mild episodes and only 6 of the severe illness. Complications in the latter sub-group of cases, included pseudocyst (4 cases) and pancreatic abscess in 2 cases. Three of the patients died, giving overall mortality of 4.2%. Two deaths were directly related to the severe pancreatitis and one was due to massive vomiting and aspiration. CONCLUSION: It is concluded that 1. Cholelithiasis is the most frequent cause of acute pancreatitis; 2. Biliary pancreatitis occurred more frequently in elderly females and was associated with higher mean levels of lactic dehydrogenase and aspartate transaminase as compared to patients with non-biliary causes. In Jizan, acute pancreatitis presents in a mild form with low morbidity and mortality, irrespective of the cause. PMID- 12590282 TI - Detection, staging and clinical implications of renal cell carcinoma. AB - OBJECTIVE: To compare the epidemiologic, clinical, and pathologic characteristics of symptomatic and incidental renal cell carcinoma (RCC) in Jordan, with some emphasis on age distribution and its potential effect in defining groups of patients that may benefit from early detection programs. We compared our results with the published western figures. METHODS: Records of 119 patients with renal tumors diagnosed during the period January 1992 to December 2001 at Jordan University of Science and Technology, Irbid, Jordan were reviewed. Age, gender, discovery circumstances of the tumor, radiologic and biologic workup, treatment, tumor node metastasis classification, and histologic features of the tumor were analyzed. RESULTS: The mean patient age was 54, and the male to female ratio was 3.4:1. The annual frequency rate for RCC per 100,000 population was 1.2 for both sexes, while for men was 1.8 and for women was 0.5. Twenty-six percent of tumors were discovered accidentally. The mean age at the time of diagnosis was not influenced by the discovery circumstances. The incidental detection group had significantly small size of tumor (5.6 cm circumference (c.f) 8.1 cm), lower stage and lower histological grading. In the symptomatic group; women have significantly lower mean size of tumors than men (5.5 cm c.f. 7.7 cm; p = 0.005; t-test), while there is no significant difference among the incidental group. A radical nephrectomy was performed in 92% of the cases, and in 8% of the cases, conservative management was adopted. CONCLUSION: The present study showed that the incidence rates of RCC in Jordan and other Middle East countries were less than most of the western countries. Malignant renal tumors in Jordan tend to affect people at a remarkably young age. This is mostly a reflection of the high proportion of young people in this country. This high portion of young people may explain the low incidence as kidney cancer is known to increase with age. Significant numbers of RCC were detected incidentally with lower pathological stage and grade. Subsequently these clinically and histologically less aggressive lesions lead to better survival and decreased recurrence. These data efforts should be directed to the development of a screening protocol to detect these lesions early, so that they may be prevented from progressing to the point when symptoms are apparent and prognosis becomes worse. PMID- 12590281 TI - Mouth and genital ulcerations in the community. AB - OBJECTIVE: To estimate the prevalence of mouth and genital ulceration in the community and its relationship to rheumatic diseases. METHODS: A house to house survey was carried out over a period of 18 months extending from September 1993 to February 1995, on 5,894 individuals utilizing detailed questionnaires on symptoms of musculoskeletal diseases and associated symptoms including history of mouth, genital ulcers, eye symptoms, and skin changes. After the initial phase, a 2nd phase was completed by trained nurses and paramedical staff, the purpose of which was to explore in detail, the history of those responding positively to any of the questions in phase one. The 3rd stage was conducted by general practitioners and rheumatologist to interview and examine those identified in phase 2 at King Fahad Hospital, Buraidah, Kingdom of Saudi Arabia. RESULTS: We identified 7 (0.1%) cases of genital ulceration, 43 (0.7%) cases of mouth ulceration, 263 (4.5%) cases of reddish eye or blurred vision, and 124 (2.1%) cases of skin rashes. Only 2 cases had both mouth and genital ulceration in addition to eye disease, skin rash and musculoskeletal pain. There was significant correlation between mouth ulcers and skin rash, eye symptoms, genital ulcerations, male sex, single status, weight loss and backpain. CONCLUSION: The vast majority of mouth and genital ulcers are not associated with a known rheumatic disease. PMID- 12590283 TI - Chlamydia trachomatis infections among Yemeni school pupils in relation to environmental conditions. AB - OBJECTIVE: This study is aimed at establishing the prevalence of Chlamydia trachomatis infections among school pupils in Sana'a, Republic of Yemen, and to explore the association of infection with environmental and social factors. METHODS: A total of 787 school children, 529 boys and 258 girls, were randomly selected for enrollment in this study. Four hundred and twenty-nine were from 4 schools in Sana'a city, and 358 from 3 rural schools around Sana'a, Republic of Yemen. Questionnaire forms were filled in for each child to investigate environmental and social factors. Sera were tested for anti Chlamydia trachomatis IgG antibodies using enzyme-linked immunosorbent assay CTM-IgG . RESULTS: An overall rate of infection of 45.9% was determined. The rate of infection among rural pupils (73.2%) was higher (P<0.0001) than that among urbanones (23.1%). The rate of infection was found significantly (P<0.001) inversely correlated with age of the pupils. Environmental factors which were found to influence the infection rate were; rural residence, unplastered walls, mud floor, lack of stand pipe water, lack of latrine and presence of animals within dwelling with odds ratio of 9.1, 6.3, 6.1, 5.2, 3.7, 3.5 and 1.7. Also, the male sex and illiteracy of the parents has been found to be risk factors for infection. CONCLUSION: Prevalence of Chlamydia trachomatis infection was found to be high. The prevalence correlates inversely with age. Rural residence, environmental conditions and social factors were risk factors for infections. Yemen could be identified as a trachoma endemic area, which should be targeted by the control programs. PMID- 12590284 TI - Congenital acinar dysplasia. Familial cause of a fatal respiratory failure in a neonate. AB - Pulmonary hypoplasia is a rare cause of pulmonary insufficiency, and has a significant rate of morbidity and mortality among affected infants. In most cases, pulmonary hypoplasia is secondary to underlying abnormalities. These may include space occupying lesions, as in infants with congenital diaphragmatic hernia; malformation of chest wall resulting in a small thoracic cavity; severe and prolonged oligohydramnios; and neuromuscular disorders, which prevent normal fetal chest expansion. All lead to poor lung development. Primary pulmonary hypoplasia as a result of congenital acinar dysplasia is exceedingly rare and is diagnosed by exclusion of all known etiologies of secondary pulmonary hypoplasia. PMID- 12590285 TI - Spontaneous resolution of infantile nephrotic syndrome. AB - Congenital nephrosis is a rare disease with universally accepted poor prognosis; despite developments in the pathogenesis and management, these children are always a challenge to the caregivers and families. Reported here, is a case of a 6-month-old female infant with infantile nephrotic syndrome, who went into spontaneous resolution within a week without therapy and remained asymptomatic for the following 2 years. PMID- 12590286 TI - Unusual radiological presentation of renal cell carcinoma. AB - We present an unusual case of renal cell carcinoma in a 59-year-old Saudi male less than 3 cm in size showing a pelvicalyceal filling defect on excretory urography and retrograde pyelography. Renal stones and blood clots were excluded by ultrasound and computerized tomography scanning. A urothelial tumor was initially diagnosed; finally surgery revealed a papillary renal cell carcinoma. PMID- 12590287 TI - Sickle cell disease and cystic fibrosis. AB - Simultaneous occurrence of both cystic fibrosis and sickle cell disease in the same patient is a rare phenomenon and has occurred only in 3 reports in the world literature. This phenomenon has never been described before in Arab population. In this report, we describe the first reported case of both diseases in one patient in the Arab population, and discuss its complications and review the literature in this aspect. PMID- 12590288 TI - Silicosis in a housewife. AB - Silicosis is an occupational respiratory disease caused by inhaling respirable crystalline silica dust. We report the case of silicosis in a 50-year-old woman that presented with pulmonary fibrosis and later developed pulmonary tuberculosis. Her condition is believed to be due to inhalation of dust produced by scrubbing a silica rich surface during usual daily housework. PMID- 12590289 TI - Spontaneous cure of an oropharyngeal hamartoma. PMID- 12590290 TI - Laparoscopic diagnosis and management of splenogonadal fusion. PMID- 12590291 TI - Paradoxical response in tuberculosis and reversal reaction in leprosy. Different terminology for same immunology? PMID- 12590292 TI - The age of onset of psoriasis and its relationship to smoking habits and stressful life events. PMID- 12590293 TI - Deficiences of physical examination among medical students. PMID- 12590294 TI - Behind the screen. PubMed intricacies. PMID- 12590295 TI - Troponin 1 in acute myocardial infarction. PMID- 12590296 TI - Malaria parasitemia during delivery. PMID- 12590297 TI - Autopoiesis: a review and a reappraisal. AB - The aim of the paper is to review critically the notion of autopoiesis as presented by Maturana and Varela. In particular, recognizing that there are difficulties in obtaining a complete and clear picture from the primary literature, an effort is made to present a coherent view-also based on many years of personal contact with Francisco Varela. The paper begins with a few historical notes to highlight the cultural background from which the notion of autopoiesis arose. The basic principles of autopoiesis as a theory of cellular life are then described, emphasizing also what autopoiesis is not: not an abstract theory, not a concept of artificial life, not a theory about the origin of life-but rather a pragmatic blueprint of life based on cellular life. It shown how this view leads to a conceptually clear definition of minimal life and to a logical link with related notions, such as self-organization, emergence, biological autonomy, auto referentiality, and interactions with the environment. The perturbations brought about by the environment are seen as changes selected and triggered by the inner organization of the living. These selective coupling interactions impart meaning to the minimal life and are thus defined by Maturana and Varela with the arguable term of "cognition". This particular view on the mutual interactions between living organism and environment leads these authors to the notion of "enaction", and to the surprising view that autopoiesis and cognition are two complementary, and in a way equivalent, aspects of life. It is then shown how cognition, so defined, permits us to build a bridge between biology and cognitive science. Autopoiesis also allows one to conceive chemical models of minimal cellular life that can be implemented experimentally. The corresponding work on "chemical autopoiesis" is then reviewed. The surprising impact of autopoiesis in the social sciences ("social autopoiesis") is also briefly discussed. This review also comments on why the theory of autopoiesis had, and still has, a difficult time being accepted into the mainstream of life-science research. Finally, it is pointed out that the new interest in system biology and complexity theories may lead to a reappraisal of autopoiesis and related notions, as outlined also by other authors, such as Tibor Ganti and Stuart Kauffmann. PMID- 12590298 TI - "Venom" of the slow loris: sequence similarity of prosimian skin gland protein and Fel d 1 cat allergen. AB - Bites inflicted on humans by the slow loris (Nycticebus coucang), a prosimian from Indonesia, are painful and elicit anaphylaxis. Toxins from N. coucang are thought to originate in the brachial organ, a naked, gland-laden area of skin situated on the flexor surface of the arm that is licked during grooming. We isolated a major component of the brachial organ secretions from N. coucang, an approximately 18 kDa protein composed of two 70-90 amino-acid chains linked by one or more disulfide bonds. The N-termini of these peptide chains exhibit nearly 70% sequence similarity (37% identity, chain 1; 54% identity, chain 2) with the two chains of Fel d 1, the major allergen from the domestic cat (Felis catus). The extensive sequence similarity between the brachial organ component of N. coucang and the cat allergen suggests that they exhibit immunogenic cross reactivity. This work clarifies the chemical nature of the brachial organ exudate and suggests a possible mode of action underlying the noxious effects of slow loris bites. PMID- 12590299 TI - Indirect interactions between a phytopathogenic and an entomopathogenic fungus. AB - Slow growth in herbivores may lead to higher mortality, due to prolonged exposure to natural enemies. A number of studies has tested the 'slow-growth, high mortality' hypothesis using predators or parasitoids as natural enemies of the herbivore, but the possible role of pathogens is poorly documented. We provide evidence that phytopathogenic infection of a plant enhances the susceptibility of herbivore larvae to a generalist entomopathogen. Larvae of the mustard leaf beetle Phaedon cochleariae grow more slowly when feeding on Chinese cabbage leaves infected by the phytopathogenic fungus Alternaria brassicae than larvae feeding on healthy leaves. Treatment of such larvae feeding on diseased plants with an LD(50) of the entomopathogenic fungus Metarhizium anisopliae resulted in 100% mortality, compared with a mortality rate of 54% in the control larvae feeding on uninfected leaves. This is the first demonstration of an interaction between a phyto- and an entomopathogenic fungus. PMID- 12590300 TI - Inverse correlation between mitochondrial size and metabolic competence: a quantitative cytochemical study of cytochrome oxidase activity. AB - Mitochondria are topologically closed bilayered systems where the synthesis of adenosine triphosphate (ATP) from adenosine diphosphate (ADP) and inorganic phosphate occurs via oxidative phosphorylation. The ordered architecture (and its extension) of the mitochondria (i.e. inner membrane, outer membrane and cristae) constitutes a critical topographic arrangement for their energy-providing mechanisms. Thus, quantitative estimations of the ultrastructural features of organelles preferentially stained by means of function-related cytochemical reactions reliably report on their potential to supply adequate amounts of ATP. On the basis of this rationale, we carried out a computer-assisted cytochemical study of cytochrome oxidase (COX) activity on mitochondria of different size in the cerebellar cortex of adult rats. The total intra-mitochondrial area of the cytochemical precipitates (CPA)/mitochondrion, the area (MA) and the longer diameter (F(max)) of COX-positive organelles were measured. The ratio (R): CPA/MA was also calculated and referred to as the percentage of mitochondrial inner membrane area involved in COX activity. The regression analysis of R vs MA showed a significant inverse correlation (r=-0.905). The fourfold increase in MA from quartiles I to IV was matched by increases in F(max) and CPA, respectively, but it was also related to a 25% decrease in R. By matching quantitative cytochemical estimations of COX activity within mitochondria with the morphometric assessment of their ultrastructural features, the present study correlates size to the metabolic competence of COX-positive organelles. Quantitative cytochemistry of COX activity is currently regarded as a reliable marker of cellular metabolism; thus our findings support the hypothesis that enlargements in size are inversely correlated with the mitochondrial metabolic competence. PMID- 12590301 TI - Symbiotic bacteria in hornet pupal silk. AB - The silk weave spun by hornet larvae before undergoing pupal metamorphosis is composed of fibers and sheets, both containing symbiotic bacteria. The bacteria are secreted from the silk gland and are glued to the secreted silk, which is made up of amino-acid polymers. In the dark, it possesses at first an electric current amounting to several hundred nanoamperes (nA) (i.e., a thermoelectric property), and a high electric capacitance of up to several milliFarads (mF). This electrical charge is used gradually by the developing pupa. The symbiotic bacteria penetrate through slits in the coat of the silk fibers to the core or into pockets in the sheets, where they gradually digest parts of the silk weave, thereby nullifying its mechanical properties and facilitating in due time the egress of the imago from the puparium. PMID- 12590302 TI - Adult eclosion timing of the onion fly, Delia antiqua, in response to daily cycles of temperature at different soil depths. AB - For insects pupating in the soil, the day/night temperature cycle may provide a primary time cue (Zeitgeber) for adult eclosion to occur at an appropriate time of the day. In the soil, however, the phase of temperature cycle is delayed with depth because of the low heat conductivity of the soil. Therefore pupae located deeper in the soil may compensate for the depth-dependent phase delay of Zeitgeber to avoid mistimed emergence. We examined the adult eclosion timing of the onion fly, Delia antiqua, pupating at different depths in soil and under various thermoperiods in the laboratory to determine if such compensation indeed occurs. We found that D. antiqua is able to compensate for the depth-dependent phase delay of the Zeitgeber by advancing the eclosion timing in response to the amplitude of the temperature cycle decreasing with depth. PMID- 12590303 TI - Evasive response to ultrasound by the crepuscular butterfly Manataria maculata. AB - The crepuscular nymphalid butterfly Manataria maculata was studied in Monteverde cloud forest, Costa Rica, during the dry season reproductive diapause. M. maculata has ears in the form of Vogel's organs located near the base of the forewings. Its behaviour in response to bursts of ultrasonic pulses (26 kHz, 110 dB SPL at 1 m) was condition-dependent. At dusk and dawn the sound consistently elicited evasive responses, similar to those of moths, in flying individuals. In contrast day-roosting individuals always remained motionless although they were alert to other stimuli. The daily movements between day- and night-roosts coincided in time and light intensity with the activity of insectivorous bats. This is the first reported case of ultrasonic hearing connected to evasive flights in a true butterfly (Papilionoidea). It strongly supports the idea that echolocating bats were involved in the evolution of hearing in butterflies. PMID- 12590304 TI - The use of olfaction in the foraging behaviour of the golden-mantled flying fox, Pteropus pumilus, and the greater musky fruit bat, Ptenochirus jagori (Megachiroptera: Pteropodidae). AB - Double-choice experiments with three adult males of the little golden-mantled flying fox, Pteropus pumilus, and ten adult greater musky fruit bats, Ptenochirus jagori (both Megachiroptera: Pteropodidae), demonstrate that they are able to discriminate accurately between an empty dish and a dish containing fruits of one of several species by odour alone. Tests were run using fruits of six fruit species for Pteropus pumilus and five fruit species for Ptenochirus jagori. The fruit species used are known to be consumed in the wild by Ptenochirus jagori and are, with two exceptions, species of the natural rain-forest habitat. This is the first study to show that fruit bats are also able to assess the ripeness of a fruit exclusively by its odour. The bats preferred ripe over unripe fruits of the same species. Thus, both Pteropus pumilus and Ptenochirus jagori can not only locate fruits by their odour but can also discriminate between ripe and unripe fruits of the same species by olfaction. The results confirm and expand earlier findings on the role of olfactory cues in the orientation of foraging pteropodids. PMID- 12590305 TI - Colony size affects division of labour in the ponerine ant Rhytidoponera metallica. AB - In theory, larger colonies of social insects should have greater colony organisation. While inter-specific comparative studies provide support for this idea, there is little direct intra-specific evidence. We investigated differences in task specialisation between large (>450 workers) and small (<80 workers) colonies of the ponerine ant Rhytidoponera metallica. Observations of individually marked young or old workers revealed greater task specialisation in large colonies. Age polyethism was detected in large but not small colonies. In large colonies, old workers spent significantly more time foraging than young workers did, while young workers spent more time caring for brood. In small colonies, young and old workers spent a similar amount of time foraging and caring for brood. This difference in task allocation patterns in large and small colonies was associated with a difference in contact rates between workers. Workers in small colonies have a lower contact rate between nestmates and a greater variability in time between contacts than workers from large colonies. PMID- 12590306 TI - [Epidemiology of food allergies]. AB - Difficulties resulting from diagnosis of food allergies show an impact on the validity of epidemiological data, which are mainly based on questionnaire data or results from allergy tests for evidence of specific IgE antibodies. The frequency of self-reported adverse food reactions in population based studies ranges between 5% and 33% with women reporting a higher frequency of such reactions. Only two epidemiological studies have performed double blind, placebo-controlled oral food challenge tests. Extrapolating these results, the prevalence of food allergies is estimated at 2% within the general population. An epidemiological questionnaire-based study in France reports on 3.5% of the general population suffering from food allergies. According to studies of the Allergy Ward in Zurich, in 46-60% of all cases skin and mucosa are predominantly affected, with oral allergy syndrome and urticaria being the most frequent manifestations. A food allergy is significantly associated with different manifestations of atopy and predominantly to hay fever, atopic eczema, urticaria and asthma. PMID- 12590307 TI - [Cross-reactive allergen clusters in pollen-associated food allergy]. AB - Immediate symptoms caused by allergens without previous primary sensitization are commonly based on cross-reactive IgE antibodies. They are responsible for pollen associated food allergies, i.e., fruit allergy in cases of birch pollen allergy or allergy to celery and spices in cases of mugwort pollen allergy. Similar structures between the major allergen of birch pollen (Bet v 1) and a variety of pathogenesis-related proteins from the same family (PR-10), abundant in hazelnuts, fruits, and vegetables, have been well established. Other candidates are profilins, ubiquitous panallergens with little clinical relevance, and stable lipid transfer proteins, responsible for systemic reactions predominantly in Mediterranean countries. Depending on stability, dose, and resorption of these proteins, clinical symptoms are limited to the oro-pharyngeal cavity or develop systemically far from the site of allergen exposure. Clinical diagnosis is based on a positive case history with corresponding allergic sensitization to pollen allergens. Targeted skin testing with native products (i.e., prick to prick test with fresh fruits) appears to be superior for unstable allergens compared to commercial extracts. Individuals should be familiar with cross-reactive patterns; however, allergen avoidance is only recommended in cases of clinical symptoms. PMID- 12590308 TI - [Relevance of food allergies and intolerance reactions as causes of urticaria]. AB - Adverse reactions to food can be identified as underlying causes in various urticarial diseases. Genuine IgE-dependent allergic reactions to food are known to play a role in acute urticaria (especially in young children), in some cases of exercise-induced urticaria, and in contact urticaria. Intolerance reactions to certain foods (pseudoallergens) are recognized causes of chronic urticaria. A thorough and detailed history, keeping a urticaria diary, quantification of specific serum IgE, and skin tests as well as elimination and provocation tests can help to identify food allergy or intolerance as causes of urticaria. In urticaria patients diagnosed with food allergy or intolerance, appropriate diets will usually result in the complete remission of disease. PMID- 12590309 TI - [Ocular complications of adjuvant interferon therapy for malignant melanoma: a review]. AB - BACKGROUND AND OBJECTIVE: In dermatology, interferon Alfa 2 is used in adjuvant therapy of melanoma (stage IIa/b) as well as in treatment of cutaneous lymphoma or melanoma (stage III or higher). In the past 2 years, several case reports have described interferon-induced ocular side effects, including sudden loss of vision and ischemic retinopathy. We wondered if incidence and prognosis of ocular complications were elevated in patients receiving an adjuvant treatment of melanoma with interferons. PATIENTS/METHODS: We analyzed 1334 reports of adverse effects of interferons, which had been spontaneously reported in Germany since 1990. RESULTS: 8.4% of all reported adverse effects of interferons were ocular. More than the half of these patients developed significant visual loss including retinal ischemia. CONCLUSIONS: These data underscore the importance to inform patients concerning ocular adverse effects and emphasize the need to monitor the retina during adjuvant interferon therapy. PMID- 12590310 TI - [Clinical course and therapy of Langerhans cell histiocytosis in children and adults]. AB - The Langerhans cell histiocytosis is a rare disease with a clear preference of infancy and an incidence of 0.2 to 1.0/100,000 children occurring worldwide. In 60 to 70% of the cases the first manifestation is before the second year of age. The disease prefers the male sex in a male to female relation of 2:1. Occurrence in adult age is very rare. Up to 10% of the cases are congenital. Both the etiology and the pathogenesis of the disease are unknown. The clinical presentation is extremely variable and covers a wide spectrum from localized involvement with a favorable prognosis to disseminated, malignant and foudroyant course. We report on ten patients, six infants and children and four adolescents and adults. We point out the very variable morphologic manifestations, the different disease course and the therapeutic options. PMID- 12590311 TI - [Palmoplantar vesicular lesions in childhood]. AB - BACKGROUND AND OBJECTIVE: Palmoplantar vesicles in children have various underlying causes, requiring different therapies. We evaluated the most common underlying diseases and determined simple criteria for differentiation. PATIENTS/METHODS: Within a two years period all children up to 14 years of age who presented with acral vesicles were included in this study. RESULTS: The most common causes of acral vesicles in a group of 32 patients, were dyshidrotic eczema with (n=11) or without atopic diathesis (n=11) and scabies (n=7). Rarely, the cause of vesicular lesion was tinea (n=2) or infantile acropustulosis (n=1). While dyshidrotic eczema was a disease of late childhood, palmoplantar lesions caused by scabies developed in younger children up to the age of 4 years. Scabies in contrast to infantile acropustulosis tend to present with more generalized lesions, not being restricted to acral location. Dyshidrotic eczema revealed lesions bilaterally and in case of atopy, additional body areas were involved. Unilateral presentation was a clue for tinea. CONCLUSIONS: Acral vesicles in childhood can be diagnostically discriminated by the age of the patient and the distribution of the lesions. PMID- 12590312 TI - [Sarcoidal allergic contact dermatitis due to palladium following ear piercing]. AB - Granulomatous tissue reactions due to jewelry made of gold, silver, nickel and palladium are rare but nevertheless have been known for a long time. A female patient developed nodular infiltrates after having been pierced with ear stickers containing palladium. A contact allergic reaction could be demonstrated as the underlying cause by inducing similar histological changes following patch testing with palladium. PMID- 12590313 TI - [Complications of pseudoainhum]. AB - A 39 year old man presented with recurrent episodes of leukocytoclastic vasculitis and a large ulcer at the distal part of the stump of his right leg. Since birth the patient showed a clinical picture of pseudoainhum as part of the amniotic band syndrome (Simonart syndrome). Radiologic examination of the right foot showed only an hypoplastic talus and calcaneus which had been designated as a "stump":, which have been detected by x-ray. On both upper limbs there were missing phalanges, syndactyly and some constrictive fibrotic bands. Since the ulcer worsened with therapy, the patient underwent the amputation of the stump just below under the knee. Since then no further vasculitic lesions have been observed. We consider this case remarkable both because pseudoainhum is so rare and because of these equally unusual complications. PMID- 12590314 TI - [From the New World. Louis A. Duhring and dermatitis herpetiformis]. AB - Louis Adolphus Duhring (born December 23, 1845) died in Philadelphia, the city of his birth and life work, on May 8, 1913. After medical school, he studied for two years in dermatology departments in London, Paris and Vienna. As a student in Vienna, he was under the tutelage of Ferdinand von Hebra, when the brilliant clinician and teacher was at the zenith of his fame. Duhring's series of 18 papers--published between 1884 and 1891--gave him an important and recognized position as one of the leading dermatological thinkers of the world. These publications described the skin disease which he named "dermatitis herpetiformis" (Duhring's disease). His assertion that dermatitis herpetiformis was a distinct dermatological disease process aroused at first considerable opposition. Moriz Kaposi, at that time the doyen of the Vienna school of dermatology, attacked his views with vigor and tenacity. Louis A. Duhring--this modest, unassuming, quiet working, gentlemanly physician--made monumental contributions to American dermatology and played a significant role in establishing Philadelphia as one of the great centers of dermatologic activity. PMID- 12590315 TI - [Vaccination with human "papillomavirus-like particles"]. PMID- 12590316 TI - Radiation-induced bystander effects. Mechanisms, biological implications, and current investigations at the Leipzig LIPSION facility. AB - BACKGROUND: The bystander effect is a relatively new area of radiobiological research, which is aimed at studying post-radiation changes in neighboring non hit cells or tissues. The bystander effect of ionizing irradiation is important after low-dose irradiation in the range of up to 0.2 Gy, where a higher incidence of stochastic damage was observed than was expected from a linear-quadratic model. It is also important when the irradiation of a cell population is highly non-uniform. OBJECTIVE: This review summarizes most of the important results and proposed bystander effect mechanisms as well as their impact on theory and clinical practice. The literature, in parts contradictory, is collected, the main topics are outlined, and some basic papers are described in more detail. In order to illustrate the microbeam technique, which is considered relevant for the bystander effect research, the state of the Leipzig LIPSION nanoprobe facility is described. RESULTS: The resistance of a radiation-induced bystander effect is now generally accepted. The current state of knowledge on it is summarized here. Several groups worldwide are working on understanding its different aspects and its impact on radiobiology and radiation protection. CONCLUSION: The observation of a bystander effect has posed many questions, and answering them is a challenging topic for radiobiology in the future. PMID- 12590317 TI - [Radiochemotherapy with gemcitabine and cisplatin in pancreatic cancer -- feasible and effective]. AB - BACKGROUND: Concomitant radiotherapy and chemotherapy with gemcitabine appears to be a promising tool for the treatment of pancreatic cancer since gemcitabine -- applied as single or combination therapy -- proved to have better efficacy in pancreatic cancer than 5-FU containing schemes and furthermore offers radiosensitizing potential. In the present paper our pilot data of concomitant and sequential chemoradiation with gemcitabine and cisplatin are presented. PATIENTS AND METHODS: A total of 57 patients (f/m 23/34) with pancreatic cancer was treated, of whom 33 patients had irresectable tumors, 19 patients following resection (R1 and/or pN+) and five patients with local recurrent disease. Radiotherapy was delivered in 25 fractions up to a total dose of 45.0 Gy specified according to ICRU reference point (50 patients, 1.8 Gy/fraction) respectively 50.0 Gy to gross tumor volume (seven patients; 45.0 Gy in locoregional lymphatic pathways; 2.0/1.8 Gy/fraction). Concomitant with radiotherapy cisplatin (30 mg/m(2)) and gemcitabine (300 mg/m(2)) were applied on days 1, 8, 22 and 29. After simultaneous chemoradiation two sequential cycles gemcitabine and cisplatin (1000 mg/m(2) and 50 mg/m(2) d 1, 15) were applied. RESULTS: With a median follow-up of 8.2 months the median survival time was 14.8 months (irresectable patients: 10.3 months, postoperative patients, 15.1 months). Within 33 irresectable patients 19 and four partial and complete remissions, respectively, were observed. In 14 patients a secondary resection was possible. Using leveled antiemetics with ondansetron and dexamethasone no gastrointestinal toxicities grade III or IV were observed. Hematologic toxicities were the most grave side effects (leukocytopenia III/IV in 29/five patients and thrombocytopenia III/IV in 21/eight patients), however with minor clinical relevancy (one neutropenic infection, one thrombopenic epistaxis). CONCLUSION: The presented treatment scheme using concomitant and sequential gemcitabine and cisplatin with radiation is feasible with justifiable side effects. To evaluate the promising remission and survival rates, randomized trials of neoadjuvant and primary chemoradiation are started. PMID- 12590318 TI - Chemoradiation of unresectable pancreatic carcinoma: impact of pretreatment hemoglobin level on patterns of failure. AB - AIM: To evaluate, in patients with locally advanced pancreatic carcinoma undergoing concomitant chemoradiation, the impact of pretreatment hemoglobin (Hb) concentration on the outcome in terms of clinical response, local control, metastasis-free survival, disease-free survival, and overall survival. PATIENTS AND METHODS: 30 patients undergoing concomitant chemoradiation (5-fluorouracil [5 FU], 1,000 mg/m(2)/day, continuous i.v. infusion days 1-4 of radiotherapy) and external beam radiotherapy (50.4-59.4 Gy) were divided into two groups based on pretreatment median Hb value (11.5 g/dl). The potential prognostic factors examined besides Hb concentration were: tumor site (head vs body-tail), sex (female vs male), cN (cN0 vs nC1), dose of external beam radiotherapy (50.4 Gy vs 59.4 Gy), presence of jaundice at diagnosis (yes vs no), weight loss at diagnosis (> or = 5 kg vs < 5 kg), epigastric-lumbar pain at diagnosis (yes vs no), maximum tumor diameter (< 40 mm vs > or = 40 mm). RESULTS: Pretreatment Hb ranged between 9.6 and 15.0 g/dl. No statistically significant differences were observed as for clinical response and local control between patients with an Hb < or = 11.5 g/dl and those with an Hb > 11.5 g/dl. Metastasis-free survival was 5.1 months in patients with an Hb < or = 11.5 g/dl and 10.7 months in patients with an Hb > 11.5 g/dl (p = 0,010). Median actuarial disease-free survival was 5.1 and 10.2 months in patients with an Hb < or = 11.5 and > 11.5 g/dl, respectively (p = 0.026). Median actuarial overall survival was 7.5 and 10.3 months in patients with an Hb < or = 11.5 and > 11.5 g/dl; respectively (p = 0.039). On multivariate analysis, Hb concentration at diagnosis was the only factor prognostically correlated with metastasis-free survival (p = 0.026), disease-free survival (p = 0.032), and overall survival (p = 0.048). CONCLUSION: In a group of patients with locally advanced pancreatic carcinoma treated with chemoradiation, a significant correlation was observed between pretreatment Hb levels and metastasis-free survival, disease-free survival, and overall survival. PMID- 12590319 TI - Significant increase in residual DNA damage as a possible mechanism of radiosensitization by gemcitabine. AB - PURPOSE: To investigate the effect of gemcitabine (dFdC), a promising radiosensitizing nucleoside analog, on the induction and repair of DNA double strand breaks (dsbs) after ionizing radiation (RT) in a pancreatic tumor cell line. MATERIAL AND METHODS: BxPC3 pancreatic tumor cells were treated using different concentrations of gemcitabine with and without subsequent irradiation. DNA dsbs were detected by constant-field gel electrophoresis under neutral conditions. RESULTS: With the addition of gemcitabine (0.5-1,000 micromol/l for 2 h prior to RT) to RT (0-75 Gy), a considerable and dose-dependent increase of remaining DNA damage after 24 h (5.4-fold for 0.5 micromol/l dFdC, 12.2-fold for 1,000 micromol/l dFdC at 25 Gy) was noted. Enhancement factors were inversely correlated with increasing X-ray dose (7.8-fold for 0.5 micromol/l at 1 Gy decreasing to 1.6-fold at 75 Gy). Conversely, the induction of DNA dsbs was not affected. Gemcitabine alone lead to a slight increase of initial DNA dsbs and only a modest elevation of residual DNA damage. CONCLUSION: These findings strengthen the hypothesis of DNA repair inhibition as a major mechanism of radiosensitization by gemcitabine. PMID- 12590320 TI - Effects of serum starvation on radiosensitivity, proliferation and apoptosis in four human tumor cell lines with different p53 status. AB - PURPOSE: The effects of serum starvation on radiation sensitivity, cell proliferation and apoptosis were investigated with particular consideration of the p53 status. MATERIAL AND METHODS: Four human tumor cell lines, Be11 (melanoma, p53 wild-type), MeWo (melanoma, p53 mutant), 4197 (squamous cell carcinoma, p53 wild-type) and 4451 (squamous cell carcinoma, p53 mutant), were used. After the cells had been incubated in starvation medium (0.5% FCS) for 1-6 days, changes in cell cycle distribution, induction of apoptosis and necrosis, and changes in radiation sensitivity were assessed by two-parameter flow cytometric measurements of DNA-dye-exclusion/Annexin V binding, and a conventional colony assay, respectively. RESULTS: p53 wild-type cell lines showed a decrease in the BrdU labeling index and an increase in the apoptotic cell frequency in starvation medium. p53 mutant cell lines showed a decrease in the BrdU labeling index but no evidence of apoptosis. These cells went into necrosis instead. The radiation sensitivity was increased in 4451 and slightly decreased in Be11 and 4197 in starvation medium. CONCLUSION: These data suggest a functional involvement of p53 in starvation-induced G1-block and apoptosis in tumor cells. Altered radiosensitivity after culture in starvation medium seemed to be explained at least in part by the starvation-induced G1-block. The frequency of starvation-induced apoptosis or necrosis was not correlated with radiation sensitivity. PMID- 12590321 TI - Logical checking function increases the accuracy of data entry in the patterns of care study. AB - BACKGROUND: For clinical studies, the use of a databases combined with a computer is becoming popular for data capturing. Commercially available database software has a logical checking function, which can restrict the type of data and their range. The study presented here quantitatively and prospectively evaluated the effectiveness for data management with this function using the database of Patterns of Care Study (PCS) in Japan. PATIENTS AND METHODS: The first external audit was performed from September 1998 to March 1999 using the PCS database without the function, and the second from April to September 1999 with the function. We analyzed the numbers and patterns of erroneous data for these two periods obtained with and without the function. RESULTS: In the database without the function, the number of incorrect data was 2,812 out of 596,186 (0.47%), while the corresponding number with the function was significantly reduced to 161 out of 533,656 (0.03%, p < 0.0001). CONCLUSIONS: The data accuracy was improved by using the computer database more extensively than the conventional case report form. By adding the logical checking function to the PCS database, significant improvement in the accuracy of data entry was accomplished. This indicates the potential for increasing the accuracy of data capturing with the aid of a computer. PMID- 12590322 TI - Remote afterloading for neutron brachytherapy using californium-252. AB - BACKGROUND: Despite a pronounced technical process attained in radiotherapy of malignant neoplasms, no remarkable improvement in the treatment results has been achieved. The reason for this stagnation is the interaction between tumor cell and photon radiation. Tumor resistance against photon bombardment can be broken down by applying high linear energy transfer (LET) radiation-based treatment. The discovery of californium-252 ((252)Cf) nuclide, a source of gamma neutron radiation, established a precondition for using neutrons in tumor brachytherapy. The design of a remote afterloading device using (252)Cf sources remains an unsolved problem. MATERIAL AND METHODS: The afterloading device has been designed as a stationary radiator which is composed of three mutually interconnected units: 1. the control and drive unit consisting of a control computer and a motor driven bowden system carrying the (252)Cf source; 2. the source which is housed in a watertight concrete vessel-storage strong room, situated in the ground at a depth of 25 cm beneath the patient's bed; 3. the afterloading application module installed in the irradiation room. RESULTS: Remote afterloading allows simple, inexpensive and highly efficient radiation protection and work safety for the operating personnel. The sources may be moved arbitrarily during treatment with a position accuracy of 0.5-1.0 mm within a distance of 520 cm from the source storage position in the strong room to the application position. Both afterloading systems' unused indexer outputs are protected electronically and mechanically against any unintentional movement of the source outside the application tubes. CONCLUSION: The technologic concept of the present automatic afterloading device for neutron brachytherapy represents a possible option from the range of conceivable design variants, which - while minimizing technologic and economic requirements - provides the operating personnel with optimum protection and work safety, thus extending the applicability of high LET radiation-based treatment methods in clinical practice. PMID- 12590323 TI - Hyperfractionated 192Ir brachytherapy for recurrent retroperitoneal sarcoma: a technique for delivery of local tumor boost dose. AB - BACKGROUND: Radical surgery is the treatment of first choice for retroperitoneal sarcoma. However, locoregional relapse is frequently observed leading to death in the majority of patients. The role of radiotherapy is not well defined in the management of retroperitoneal sarcoma. Yet, there is evidence that adjuvant irradiation does improve local tumor control. MATERIAL AND METHODS: In order to deliver sufficiently high radiation doses to the retroperitoneum, different techniques for application of a local tumor boost dose in addition to external beam treatment have been proposed. We present a technique of hyperfractionated (192)Ir brachytherapy (HFIR) of the tumor bed via intraoperatively implanted plastic catheters. Postoperative CT-based image-guided brachytherapy was performed. In two consecutive patients with recurrent retroperitoneal sarcoma, treatment was delivered twice daily with single doses of 1.5-2.0 Gy in 5-10 mm tissue depth up to a total dose of 18-32.5 Gy. RESULTS: HFIR of the tumor bed was easily accomplished facilitating delivery of high radiation doses to the retroperitoneum. No major late effects of treatment have been observed with a follow-up of 15 and 28 months, respectively. Details of the brachytherapy procedure are presented. CONCLUSION: HFIR via intraoperatively implanted catheters in the retroperitoneum is a technique suitable for application of a local tumor boost dose. Thus, sufficiently high doses of radiation mandatory for long-lasting local tumor control can be delivered in the tumor bed of the retroperitoneum without exceeding normal tissue radiotolerance in this unfavorable disease. PMID- 12590324 TI - Epidural metastasis in nasopharyngeal carcinoma. AB - BACKGROUND: In nasopharyngeal carcinoma, both, a short metastasis-free interval after primary treatment and the occurrence of epidural metastasis have been associated with poor prognosis. We present the clinical course of a young patient with these two conditions and review the literature. PATIENT: A 26-year-old male with stage T2N3M0 non-keratinizing carcinoma (WHO type 2) of the nasopharynx was treated with induction chemotherapy and radical radiotherapy, 6 months after documentation of a clinical complete remission, the patient experienced metastatic disease to the C7-D1 vertebral bodies associated with an epidural soft tissue mass. Since no further metastatic lesions were detectable, the patient was treated with radiotherapy alone (3 960 cGy/22 fractions). RESULTS: Treatment resulted in compete resolution of neurological and radiological signs of the disease and the patient continues to be disease-free, 32 months after salvage treatment. In a literature search, we identified 54 reported cases with long-term survival after treatment for metastatic nasopharyngeal cancer. The vast majority of them had primary tumors with undifferentiated histology and was treated with combination chemotherapy. In 25 of them, radiotherapy was given as consolidation therapy (in 19 cases for bone and in six cases for mediastinal lymph node metastases). CONCLUSIONS: Epidural metastatic disease from a nasopharyngeal carcinoma is highly sensitive to moderate doses of fractionated radiotherapy. MR imaging is essential for the detection of relevant soft tissue disease extensions within the epidural space and proper selection of the radiation target volume in vertebral metastases. In patients with nasopharyngeal carcinoma, the occurrence of a solitary epidural metastasis after a short metastasis-free interval is not incompatible with long-term survival. PMID- 12590325 TI - A predictive model for the management of community-acquired pneumonia. AB - BACKGROUND: Understanding what determines the prognosis of community-acquired pneumonia (CAP) is especially important for decisions on hospitalization and antimicrobial therapy. The objective of the present study was to compare the predictability of mortality in our patients to that of the pneumonia patient outcomes research team (PORT) study. PATIENTS AND METHODS: Data of 320 patients admitted with CAP were retrospectively evaluated and classified according to the published scheme. RESULTS: One-month mortality was 14.4%; 1-year mortality was 27.8%, two-thirds from new episodes. Univariate logistic regression risk factors for the 1-month mortality rate included leukocytosis, anemia, hypoalbuminemia, elevated blood urea nitrogen, >or= two comorbidities, tachycardia, tachypnea, acidosis, stupor, age > 65 years and high serum lactic dehydrogenase. These variables, except the last two, plus pleural effusion and bilateral infiltration were also risk factors for 1-year mortality. In the multivariate models, eight of these factors were significant risk factors, four for 1-month mortality and six for 1-year mortality. Our model for prediction of 1-month mortality had a sensitivity of 65%, specificity of 95% and accuracy of 91%. CONCLUSION: Agreement between predictions by our model and the published model was considerable, showing that most patients in the low score groups should not have been hospitalized. PMID- 12590326 TI - Antibiotic use and the prevalence of antibiotic resistance in bacteria from healthy volunteers in the dutch community. AB - BACKGROUND: Although most antibiotics are consumed in the community (80%), the relation of antibiotic consumption and resistance in the community has not been thoroughly studied. MATERIALS AND METHODS: In the present study, antibiotic use was related to the prevalence of antibiotic resistance of Escherichia coli and enterococci isolated from healthy volunteers living in the southern part of the Netherlands in 1996 and 1999. RESULTS: No change in the total antibiotic use in the Dutch community was observed between 1996 and 1999 (3542 and 3598 defined daily doses [DDD] per 1000 inhabitants/year). However, the increased fluoroquinolone use (+ 18%) and the increased prevalence of ciprofloxacin resistance from 0 to 2% is a point of concern, especially since this was accompanied by a significant shift towards higher minimum inhibitory concentration (MIC) values (p < 0.05). A significant decrease in the prevalence of vancomycin-resistant enterococci (p < 0.05) was found in addition to a significant shift towards lower MIC valued for avoparcin, a glycopeptide previously used as growth promoter in animal husbandry (p < 0.05). This was very likely due to the banning of avoparcin for this purpose from April 1997 onwards. CONCLUSION: In order to maintain the low level of antibiotic use and resistance in The Netherlands, surveillance of antibiotic resistance in nonclinical isolates in relation to antibiotic use is very important. PMID- 12590327 TI - Fascioliasis in eosinophilic patients in the Isparta region of Turkey. AB - BACKGROUND: The aim of this study was to investigate fascioliasis in a group of eosinophilic and non-eosinophilic patients in the Isparta region of Turkey. PATIENTS AND METHODS: All cases were examined for antibodies against Fasciola hepatica by the modified ES-ELISA method. Seropositive patients with fascioliasis were investigated by radiological and laboratory evaluations. RESULTS: Of the 756 eosinophilic patients, 6.1% were found to be positive for fascioliasis by ES ELISA, while the rate was only 0.9% among the non-eosinophilic group (statistically significant, p < 0.001). Among the seropositives, 19 cases were evaluated further. Ultrasonographic findings were found in 14 of the 19 cases. In six of nine cases, computed tomography findings were related to fascioliasis. F. hepatica eggs were found in ine of the 19 cases. CONCLUSION: Fascioliasis was demonstrated in the Isparta region of Turkey for the first time. The higher prevalence of fascioliasis among patients with eosinophilia indicates the importance of this sign for this infection. PMID- 12590328 TI - Barbecued chicken causing a multi-state outbreak of Campylobacter jejuni enteritis. AB - BACKGROUND: Although the microbiological safety of food has improved, food-borne disease remains a significant cause of morbidity and mortality in Europe. PATIENTS AND METHODS: We investigated an outbreak of Campylobacter jejuni enteritis attributed to chicken meat, affecting five out of six people attending a private barbecue party in Germany. Patients fell ill in Germany, in Liechtenstein and in Austria. 80% of the cases had been exposed to barbecued chicken; the case that denied having eaten chicken was the party host, who also handled all the food. Three of four patients submitting stool specimens had culture-confirmed C. jejuni infection. RESULTS: The chicken meat was purchased in the Tyrol (Austria) and originated from a flock of 55600 chickens raised in Carinthia (Austria). Caecal swabs were obtained in 7 weeks later from 22 chicken at the incriminated farm: 18 of the 22 samples yielded C. jejuni. The same day, six carcasses out of 22000 slaughtered animals from the incriminated farm were tested and all six food samples yielded C. jejuni. Outbreak-associated human isolates yielded pulsed-field gel electrophoresis patterns indistinguishable from each other and from the meat isolates, but different from four human control strains and from 13 of 16 isolates from caecal swabs. CONCLUSION: Our data show that the outbreak clone had been colonizing the slaughterhouse and was cross contaminating chickens there. The geographic mobility of people and food necessitates proper epidemiologic investigations to avoid overestimation of the proportion of sporadic occurrence of campylobacteriosis. PMID- 12590329 TI - The humoral immune response in melioidosis patients during therapy. AB - BACKGROUND: This study was undertaken to identify and quantify the class and subclass antibody responses to the culture filtrate antigen (CFA) of Burkholderia pseudomallei in melioidosis patients under long-term maintenance or eradication therapy. MATERIALS AND METHODS: Sequential sera samples from seven melioidosis patients collected between January 1992 and April 1998 were analyzed for immunoglobulin (Ig) types and IgG isotypes by ELISA using B. pseudomallei CFA. RESULTS: Melioidosis patients generated a strong IgG, IgA and IgM response to the CFA of B. pseudomallei throughout the infection and IgG1 and IgG2 were the predominant IgG istotypes produced. Although high levels of these antibodies were detected in all the seven patients, the IgG, IgG1 and IgG2 antibodies showed a consistent response and good correlation with the clinical history in all cases. CONCLUSION: This study suggests that monitoring IgG antibody or IgG1 or IgG2 isotype antibody levels to CFA in patients under maintenance or eradication antibiotic therapy may be useful as a tool to detect the status of infection and as a guideline to determine the duration of maintenance antimicrobial therapy. PMID- 12590330 TI - Prolonged application of closed in-line suction catheters increases microbial colonization of the lower respiratory tract and bacterial growth on catheter surface. AB - BACKGROUND: Microorganisms become attracted to polymer surfaces for a number of reasons including positive charge of biomaterial or concentration of nutrients on the biomaterial surface. Many bacteria additionally possess specific receptors for the interaction with extracellular host protein components that adhere on the biomaterial surface. Several authors suggest that application of closed in-line polypropylene suction catheters (CISC) in intubated patients for more than 24 h is safe and can reduce the costs associated with mechanical ventilation. Therefore, we evaluated the possible role of prolonged application of CISC to cause enhanced colonization of both the biomaterial and the lower respiratory tract. MATERIALS AND METHODS: The prospective, randomized study included 23 mechanically ventilated patients. The CISC tips, adjacent segments and tracheobronchial aspirates of each patient were examined for microbial growth. RESULTS: Application for 72 h significantly enhanced the microbial growth on the CISC tips and on the adjacent catheter segment. Usage for 3 days led to a significant increase in colonization in the lower respiratory tract. CONCLUSION: Normal saline instillation in conjunction with endotracheal suctioning may lead to a dispersion of microorganisms into the lower respiratory tract. More effective self-cleaning mechanisms are necessary to decontaminate the CISC surface after suctioning. PMID- 12590331 TI - Cervical necrotizing fasciitis: sources and outcomes. AB - BACKGROUND: Abscesses of the peritonsillar region are among the most common deep abscesses of the head and neck. However, cervical necrotizing fasciitis (CNF) associated with a peritonsillar abscess is an extremely rare condition, with only 12 well-documented cases described. PATIENTS: We reviewed and compared all 12 cases of CNF arising from peritonsillar abscesses, including our own case. CNF that developed after peritonsillar abscesses was also compared with that developing predominantly after odontogenic infection. RESULTS: The overall mortality was higher in the group with peritonsillar abscesses (33% vs 25%). CONCLUSION: It is probable that of all cases of CNF, that arising from peritonsillar abscess has the worst prognosis. PMID- 12590332 TI - Immunological response in HIV-positive patients vaccinated against tick-borne encephalitis. AB - BACKGROUND: Vaccination against infectious diseases is a recommended preventive measure in patients with impaired immunity. The aim of the study was to estimate the immunogenicity of the vaccine against tick-borne encephalitis (TBE) in HIV infected patients and to determine its safety for this group. PATIENTS AND METHODS: The TBE vaccine FSME-IMMUN-inject was tested on 29 HIV-positive patients and the response compared to that of 40 healthy controls. The vaccination protocol for the HIV-positive group was modified by the addition of a fourth dose according to a 0/1/2/9-month schedule. RESULTS: No serious adverse reactions were observed in patients with deficient immunity. A better response was obtained in HIV-infected patients with CD4 counts >or= 500/microl (55% of the patients had levels of IgG antibody > 126 VIEU/microl) than in those with CD4 counts of 200 499/microl (40%). However, the difference did not reach significance. 85% of healthy controls achieved protective antibody titers after a full course of vaccination. CONCLUSION: The correlation between post-vaccine seroconversion and CD4 lymphocyte count showed that the FSME-IMMUN-inject vaccine can be considered to be a CD4 cell-independent vaccine. The examinations carried out 1 year after the completed vaccination protocol showed maintenance of the anti-TBE response acquired after the third vaccination in healthy subjects and in HIV-infected patients. PMID- 12590333 TI - Infection with human herpesvirus type 8 and kaposi's sarcoma in a central Italian area formerly endemic for malaria. AB - BACKGROUND: To estimate the prevalence of infection with human herpes virus type 8 (HHV8) and the incidence of Kaposi's sarcoma among HHV8-infected individuals, we conducted a population-based, cross-sectional study in Latina. This area of central Italy was formerly endemic for malaria and it is now covered by a population-based cancer registry. MATERIALS AND METHODS: Residual sera samples from 200 persons (100 men and 100 women) aged 50 years or older, randomly selected from a larger population-based survey on cardiovascular diseases, were tested for antibodies against HHV8. HHV8 seroprevalence and yearly incidence rates of Kaposi's sarcoma (KS) among HHV8-positive persons were computed. RESULTS: HHV8 infection was not significantly higher in men (9.0%, 95% CI: 3.4 14.6) than in women (6.0%, 95% CI: 1.3-10.7). The estimated yearly incidence rate of KS among HHV-positive persons was twice as high in men (1 : 2200) as in women (1 : 4110). CONCLUSION: These results were comparable to those derived from geographic areas at low risk for KS where malaria was not endemic (e. g. Malta). The seemingly higher incidence of KS among HHV8-positive men suggests that different cofactors for the two sexes are likely to play a role in KS etiology. PMID- 12590334 TI - Opsonophagocytosis versus complement bactericidal killing as effectors following Neisseria meningitidis group C vaccination. AB - BACKGROUND: Opsonophagocytosis and complement-mediated Neisseria meningitidis killing after vaccination were investigated. METHODS: Twelve seronegative healthy volunteers received one dose of polysaccharide A/C vaccine and were followed for 3 years. Ex vivo serum killing rates with polymorphonuclear cells (PMN) and/or complement were performed at 0, 1.5, 6, 12, 18, 24, 30 and 36 months. RESULTS: High mean total and median bactericidal antibodies were detected over time in all subjects. Considerable reduction of the initial inoculum was obtained only in the presence of complement, with or without PMN (with significant differences compared to curves without complement) a long time after vaccination. CONCLUSION: PMN did not increase post-vaccination bacterial killing, suggesting that antibody complement-mediated killing, and not opsonophagocytosis, is the main immune effector of the vaccine protection against N. meningitidis. PMID- 12590335 TI - Optic neuritis complicating west nile virus meningitis in a young adult. AB - A case of West Nile virus (WNV) infection with meningitis and optic neuritis in a 28-year-old man is presented. The patient had a number of unusual clinical and laboratory findings that broadened the differential diagnosis. The emergence of WNV infection in southern Europe and North America calls for increased awareness of physicians to this clinical entity. PMID- 12590336 TI - Primary cutaneous Absidia corymbifera infection in a premature newborn. AB - An unusual case of a primary cutaneous Absidia corymbifera infection in a premature twin successfully treated with low doses of intravenous amphotericin B and topical natamycin is described. Epidemiological and therapeutical aspects of the case are discussed and in vitro antifungal susceptibility data are presented. PMID- 12590337 TI - A case of imported melioidosis presenting as prostatitis. AB - We report a case of melioidosis in a previously healthy Belgian man. He presented with septicemia and prostatic abscesses 1 week after a trip to Vietnam. Burkholderia pseudomallei was isolated from multiple hemocultures. He was treated successfully with intravenous ceftazidime and trimethoprim-sulfamethoxazole, followed by a per-oral maintenance therapy of amoxicillin-clavulanate with supplementary amoxicillin. There was no need for surgical drainage. This is the second reported case of melioidosis in Belgium. PMID- 12590338 TI - Intracerebral alveolar echinococcosis. AB - There are two species of the genus Echinococcus, Echinococcus multilocularis (also called alveolar hydatid) and Echinococcus granulosus, characterized by distinct growth features in humans. The main endemic regions for human alveolar echinococcosis (AE) caused by E. multilocularis are Central Europe, Russia, Turkey, Japan, China, eastern France and North America. Human echinococcosis is usually caused by an intrahepatic growth of parasitic larvae. Cerebral occurrence of E. multilocularis disease is rare, accounting for only 1% of cases, and is generally considered to be fatal. This report presents two cases of intracerebral E. multilocularis disease which occurred in two infected patients with AE pulmonary metastases. The anatomical and clinical features are discussed. Our retrospective survey would indicate that surgical treatment should be envisaged whenever possible. PMID- 12590339 TI - Advances in cereal functional genomics. PMID- 12590340 TI - Progress in maize gene discovery: a project update. AB - The Maize Gene Discovery Project (MGDP) is a 5-year NSF-funded plant genome initiative that began in 1998. The MGDP collaboration involves researchers at six universities from diverse disciplines with the common goal of discovering new maize genes and developing tools for the phenotypic characterization of maize mutants. The project utilizes several approaches: EST sequencing, cDNA microarray production, and the discovery of gene function and genomic sequence through the use of a recombinant Mu1 transposon ( RescueMu). Current achievements of the MGDP (NSF 98-72657) include the sequencing of over 120,000 maize ESTs from diverse cDNA libraries, and over 70,000 RescueMu flanking sequences, as well as the cataloguing of mutant seed and cob phenotypes of 23,000 maize ears, 6,200 families of maize seedlings, and 4,000 families of adult maize plants carrying MuDR/Mu and RescueMu insertion alleles. A consolidation of over 24,000 unique sequences from 19 libraries has been made into the first two of the planned set of four "Unigene" microarray slides. In addition, slides for four EST libraries have been produced. These microarray slides, EST clones, library plates of immortalized RescueMu bacterial cultures, and seed are all available online (http://www.zmdb.iastate.edu). The ZmDB website posts periodic assemblies of all maize EST and genomic sequences available from GenBank. ZmDB is also a portal for sequence analysis software designed to aid in gene discovery: MuSeqBox, GeneSeqer, and SplicePredictor. In addition, ZmDB contains links to other plant and genetics websites. PMID- 12590341 TI - Arabidopsis genomic information for interpreting wheat EST sequences. AB - The resources available from Arabidopsis thaliana for interpreting functional attributes of wheat EST are reviewed. A focus for the review is a comparison between wheat EST sequences, generated from developing endosperm tissue, and the complete genomic sequence from Arabidopsis. The available information indicates that not only can tentative annotations be assigned to many wheat genes but also putative or unknown Arabidopsis gene annotations can be improved by comparative genomics. PMID- 12590342 TI - Molecular characterization of a set of wheat deletion stocks for use in chromosome bin mapping of ESTs. AB - The objective of this study was molecular characterization of a set of deletion stocks and other aneuploids for use in chromosome bin mapping of ESTs in wheat. Wheat aneuploid stocks including 21 nullisomic-tetrasomic (NT), 24 ditelosomic (Dt), and 101 deletion (del) lines were screened with 526 EST clones. A total of 1,951 loci were detected by 493 informative EST clones and tagged 150 of the 159 deletion intervals or chromosome bins. Previously described deletion lines del1AS 4, del6AL-2, del6BS-6, and del7DS-6 were found to have normal chromosome constitution. The short arm deletion in del3AS-3 may be translocated from an unknown chromosome as this stock is nullisomic for the 3AS arm. Thirty-five new deletions were detected in 26 lines. Most of the new deletions occurred in terminal regions of chromosomes and probably resulted from the loss of very small terminal fragments that were difficult to detect cytologically. Eleven chromosome aberrations were also detected in two NT and five Dt lines. Overall, the chromosome bin map provides a resolution of around 28 Mb for an anchor map of a basic set of seven chromosomes of the Triticeae. Any target gene can be allocated to a specific 28-Mb bin and associated ESTs, anchored to the other Triticeae/grass maps including rice and, therefore, amenable to molecular cloning by comparative and wheat-based positional cloning methods. PMID- 12590343 TI - The wheat D-genome HMW-glutenin locus: BAC sequencing, gene distribution, and retrotransposon clusters. AB - A bacterial-artificial-chromosome (BAC) clone from the genome of Triticum tauschii, the D-genome ancestor of hexaploid bread wheat, was sequenced and the presence of the two paralogous x- and y-type high-molecular-weight (HMW) glutenin genes of the Glu-D1 locus was confirmed. These two genes occur in the same orientation, are 51,893 bp apart, and the separating DNA includes a 31,000-bp cluster of retrotransposons. A second retrotransposon cluster of 32,000 bp follows the x-type HMW-glutenin gene region. Each HMW-glutenin gene is found within a region of mainly unique DNA sequence which includes multiple additional genes including an active endosperm globulin gene not previously reported in the Triticeae family, a leucine-rich-repeat (LRR) type gene truncated at the 5' end of the BAC, a kinase gene of unknown activity, remnants of a paralogous second globulin gene, and genes similar to two hypothetical rice genes. The newly identified globulin genes are assigned to a locus designated Glo-2. Comparison to available orthologous regions of the wheat A and B genomes show rapid sequence divergences flanking the HMW-glutenin genes, and the absence of two hypothetical and unknown genes found 5' to the B-genome x-type ortholog. The region surrounding the Glu-D1 locus is similar to other reported Triticeae BAC sequences; i.e. small gene islands separated by retrotransposon clusters. PMID- 12590344 TI - The starch branching enzyme I locus from Aegilops tauschii, the donor of the D genome to wheat. AB - Analysis of DNA from Aegilops tauschii revealed that sequences hybridisable to the starch branching enzyme I (SBE I) gene were contained within a 53-kb fragment. There were at least four genes or gene fragments but of these only one appeared to encode the SBE I observed in the endosperm. Two large-insert DNA clones that encode SBE I from A. tauschii were isolated. Hybridisation analysis confirmed the presence of multiple SBE I gene type sequences within this DNA fragment of approximately 100 kb. Fluorescent in situ hybridisation (FISH) on extended DNA fibres provided further evidence of the close proximity of three of these genes. Sequence analysis was undertaken and this demonstrated that wSBE I D3, wSBE I-D2 and wSBE I-D4 genes were clustered within 27 kb of DNA; of these only wSBE I-D4 encodes the SBE I purified from the endosperm. Multiple but distinct cDNAs containing SBE I-related sequences have been reported and these could arise from the SBE I locus by different transcription/splicing regimes. PMID- 12590345 TI - The structural organisation of the gene encoding class II starch synthase of wheat and barley and the evolution of the genes encoding starch synthases in plants. AB - Wheat and barley contain at least four classes of starch synthases in the endosperm, granule bound starch synthase I (GBSSI) and starch synthases I, II and III (SSI, SSII, SSIII). In this work, SSII in barley is shown to be associated with the starch granule by using antibodies. A cDNA from barley encoding SSII and the genes for SSII from barley and Aegilops tauschii ( A. tauschii, the D genome donor to wheat) are characterised. Fluorescent in situ hybridisation (FISH) and PCR were used to localise the wheat SSII gene to the short arm of chromosome 7, showing synteny with the location of the rice SSII gene to the short arm of chromosome 6. Comparison of the genes encoding SSII of A. tauschii, barley and Arabidopsis showed a conserved exon-intron structure although the size of the introns varied considerably. Extending such comparison between the genes encoding starch synthases (GBSSI, SSI, SSII and SSIII) from A. tauschii and Arabidopsis showed that the exon-intron structures are essentially conserved. Separate and distinct genes for the individual starch synthases therefore existed before the separation of monocotyledons and dicotyledons. PMID- 12590346 TI - Homoeologous set of NBS-LRR genes located at leaf and stripe rust resistance loci on short arms of chromosome 1 of wheat. AB - Homoeologous group 1 chromosomes of wheat contain important genes that confer resistance to leaf, stem and stripe rusts, powdery mildew and Russian wheat aphid. A disease resistance gene analog encoding nucleotide binding site-leucine rich repeat (NBS-LRR), designated RgaYr10, was previously identified at the stripe rust resistant locus, Yr10, located on chromosome 1BS distal to the storage protein, Gli-B1locus. RgaYr10 identified gene members in the homoeologous region of chromosome 1DS cosegregating with the leaf rust resistance gene, Lr21, which originally was transferred from a diploid D genome progenitor. Four RgaYr10 gene members were isolated from chromosome 1DS and compared to two gene members previously isolated from the chromosome 1BS homeologue. NBS-LRR genes tightly linked to stripe rust resistance gene Yr10 on chromosome 1BS were closely related in sequence and structure to NBS-LRR genes tightly linked to leaf rust resistance gene Lr21 located within the homoeologous region on chromosome 1DS. The level of sequence homology was similar between NBS-LRR genes that were isolated from different genomes as compared to genes from the same genome. PMID- 12590347 TI - The regulation of yp3 expression in the Drosophila melanogaster fat body. AB - The regulation of the Drosophila melanogaster yolk protein genes 1 and 2 have been well characterised. Cis-acting DNA elements and trans-acting factors regulating ovarian fat body and sex-specific expression have been identified. In this paper we have analysed the regulation of yolk protein 3, which is separated from the other two genes on the X-chromosome. We have separated sex-specific control from fat body control in some constructs in transgenic flies. We propose that the organisation of the regulatory elements in yp3 differs from yp1 and yp2 for control of fat body expression and that it closely resembles the regulation of a reporter gene using Musca and Calliphora yp promoter enhancer sequences in transgenic Drosophila. PMID- 12590348 TI - Development of the nervous system in the "head" of Limulus polyphemus (Chelicerata: Xiphosura): morphological evidence for a correspondence between the segments of the chelicerae and of the (first) antennae of Mandibulata. AB - We investigated brain development in the horseshoe crab Limulus polyphemus and several other arthropods via immunocytochemical methods, i.e. antibody stainings against acetylated alpha-tubulin and synapsin. According to the traditional view, the first appendage-bearing segment in chelicerates (the chelicerae) is not homologous to the first appendage-bearing segment of mandibulates (first antenna, deutocerebrum) but to the segment of the second antenna (tritocerebrum) or the intercalary segment in hexapods and myriapods. Accordingly, the segment of the deutocerebrum in chelicerates would be completely reduced. The main arguments for this view are: (1) the postoral origin of the cheliceral ganglion, (2) a poststomodaeal commissure, and (3) a connection of the cheliceral ganglion to the stomatogastric system. Our data show that these arguments are not convincing. During the development of horseshoe crabs there is no evidence for a former additional segment in front of the chelicerae. Instead, comparison of the brain structure (neuropil ring) between chelicerates, crustaceans and insects shows remarkable similarities. Furthermore, the cheliceral commissure in horseshoe crabs runs mainly praestomodaeal, which would be unique for a tritocerebral commissure. An unbiased view of the developing nervous system in the "head" of chelicerates, crustaceans and insects leads to a homologisation of the cheliceral segment and that of the (first) antenna (= deutocerebrum) of mandibulates that is also congruous to the interpretation of the Hox gene expression patterns. Thus, our data provide morphological evidence for the existence of a chelicerate deutocerebrum. PMID- 12590349 TI - Comparative and experimental embryogenesis of Plectidae (Nematoda). AB - Comparative analysis of early embryogenesis indicates that considerable differences exist among nematode species. To better understand to what extent the well-studied development of Caenorhabditis elegans is representative for nematodes in general, we extended our earlier studies to other families of this phylum. Here we report our findings on seven species of Plectidae. We found that Plectidae embryos share a number of developmental similarities with one branch of nematodes (Secernentea), including C. elegans, but not with the other branch (Adenophorea), and thus support conclusions concerning their phylogenetic position drawn from molecular data. However, Plectidae also show developmental differences to other Secernentea, suggesting an early separation from them. Prominent characteristics of Plectidae are (1) strict left-right divisions of somatic founder cells generating a prominent early bilateral symmetry and (2) a very early start of gastrulation with immigration of a single gut precursor cell. To determine whether gastrulation with two gut precursors is crucial for C. elegans embryos, we induced it to gastrulate with a single blastomere like in Plectidae. As this alteration is compatible with an essentially normal subsequent embryogenesis, cleavage of the gut precursor before gastrulation is obviously not required. As major differences exist among nematodes concerning the potential to compensate for eliminated early blastomeres, we tested this feature in one Plectus species. We found that Plectus does not replace a lost cell but behaves like C. elegansin this respect, in contrast to our previous findings in Acrobeloides nanus, another member of the Secernentea. PMID- 12590350 TI - Parallel microtubules and other conserved elements of dorsal axial specification in the direct developing frog, Eleutherodactylus coqui. AB - Specification of the dorsal axis in commonly studied frogs, such as Xenopus laevis and Rana pipiens, depends on a microtubule-mediated movement of cytoplasm in the fertilized egg. The Puerto Rican tree frog, Eleutherodactylus coqui, has an egg that is twenty times the volume of that of X. laevis, raising the question as to whether the mechanism of dorsal axial specification is conserved in these large eggs. Fertilized eggs of E. coqui develop a transient array of parallel microtubules, similar to other frogs, but proportionately larger. The array persists after first cleavage, longer than in other frogs, and is gone by the third cleavage. Correlated with the longer life of the parallel microtubules, both 2- and 8-cell E. coqui embryos remain sensitive to gravity-mediated axial specification, a sensitivity lost in X. laevis before the 2-cell stage. Activation of the Wnt/beta-catenin pathway by injected Xwnt8 RNA causes axial formation as in X. laevis. The results indicate that elements of dorsal axial specification are conserved in E. coqui, but they occur later compared to in X. laevis. PMID- 12590351 TI - Expression of three spalt (sal) gene homologues in zebrafish embryos. AB - Three homologues of the Drosophilaregion-specific homeotic gene spalt (sal) have been isolated in zebrafish, sall1a, sall1b and sall3. Phylogenetic analysis of these genes against known salDNA sequences showed zebrafish sall1aand sall1b to be orthologous to other vertebrate sal-1 genes and zebrafish sall3to be orthologous to other vertebrate sal-3 genes, except Xenopus sall3. Phylogenetic reconstruction suggests that zebrafish sall1a and sall1bresulted from a gene duplication event occurring prior to the divergence of the ray-finned and lobe finned fish lineages. Analysis of the expression pattern of the zebrafish sal genes shows that sall1a and sall3 share expression domains with both orthologous and non-orthologous vertebrate sal genes. Both are expressed in various regions of the CNS, including in primary motor neurons. Outside of the CNS, sall1a expression is observed in the otic vesicle (ear), heart and in a discrete region of the pronephric ducts. These analyses indicate that orthologies between zebrafish sal genes and other vertebrate sal genes do not imply equivalence of expression pattern and, therefore, that biological functions are not entirely conserved. However we suggest that, like other vertebrate sal genes, zebrafish sal genes have a role in neural development. Also, expression of zebrafish sall1a in the otic vesicle, heart sac and the pronephric ducts of zebrafish embryos is possibly consistent with some of the abnormalities seen in Sall1-deficient mice and in Townes-Brocks Syndrome, a human disorder which is caused by mutations in the human spalt gene SALL1. PMID- 12590352 TI - Utilization of a particle gun DNA introduction system for the analysis of cis regulatory elements controlling the spatial expression pattern of the arylsulfatase gene (HpArs) in sea urchin embryos. AB - We applied a particle gun method to introduce DNA into fertilized sea urchin eggs for the analysis of cis-regulatory elements responsible for spatial gene expression during development. We introduced HpArs (sea urchin arylsulfatase gene) -GFP and HpArs-LacZfusion constructs into the fertilized eggs and obtained high expression levels of the fusion genes. Using this assay system, we demonstrated that a fragment of HpArs (-3,484 to +4,636) is sufficient for aboral ectoderm-specific expression, and that the region in the first intron from +406 to +1,993 contains the control elements responsible for the repression of the HpArs promoter activity in secondary mesenchyme cells. PMID- 12590353 TI - The homeobox genes of Encephalitozoon cuniculi (Microsporidia) reveal a putative mating-type locus. AB - Homeobox genes have been found in animals, fungi, and plants. Recently, the complete genomic sequence of Encephalitozoon cuniculi has become available and it was shown that this Microsporida species is related to fungi. Given this close relatedness to fungi, I have searched the genome of E. cuniculifor homeobox genes. There are 12 homeobox genes as well as one STE12 orthologue. The large number of homeobox genes when compared to the annotations, which only list one, suggests that possibly other smaller genes may have been overlooked in the published analysis and E. cuniculi may have more than 1,997 genes. Sequence and phylogenetic analyses show that the E. cuniculi homeobox genes also fall into two distinct groups, with two TALE and ten typical homeobox genes being present. Like in the mating-type locus of yeast and other fungi, one TALE and one typical homeobox are found in close proximity of each other on the chromosome, suggesting that Microsporidia also contain a mating-type locus. PMID- 12590354 TI - Relevance of MDMA ("ecstasy")-induced neurotoxicity to long-lasting psychomotor stimulation in mice. AB - RATIONALE: Although many studies have focused on the mechanisms underlying MDMA induced neurotoxicity, little is known about the subsequent long-term response to psychostimulants following exposure to a neurotoxic dose of MDMA. OBJECTIVES: We investigated the effect of pre-exposure to neurotoxic and non-neurotoxic doses of MDMA on the response of mice to the psychomotor stimulating effects of MDMA and cocaine. METHODS: To investigate MDMA-induced neurotoxicity, male Swiss Webster mice were subjected to three regimens of MDMA: i) 40 mg/kg x 2, ii) 30 mg/kg x 2, and iii) 15 mg/kg x 2 for 2 days. On day 5 following the last exposure to MDMA, the levels of dopaminergic and serotonergic markers were determined. For the behavioral experiments, mice received either a single injection of 10 mg/kg MDMA [MDMA(L)] or one of the following doses of MDMA: 30 mg/kg x 2 or 15 mg/kg x 2 for 2 days [MDMA (H)]. A third group received saline as a control. On day 5 after the last pretreatment injection, the first MDMA (10 mg/kg) challenge was given, and on day 12, cocaine (20 mg/kg) was administered. Subsequently, mice were re challenged with MDMA on days 35, 50 and 80, after which locomotor activity was monitored by infrared beam-interrupts. On day 83, mice were killed to detect the levels of dopaminergic and serotonergic markers. RESULTS: MDMA-induced mortality and depletion of dopaminergic and serotonergic markers were dose-dependent. MDMA (H) mice endured a sensitized response to MDMA challenge from days 5 through 80, e.g. a persistent 3-fold increase in locomotor activity compared to the response of mice that were not pretreated with a neurotoxic dose of MDMA. The depletion of DAT and 5-HTT binding sites was sustained throughout this time period (64-68% of control). The MDMA (L) mice showed a sensitized response to MDMA only on day 5. Both MDMA (L) and MDMA (H) mice were sensitized to the cocaine challenge. CONCLUSIONS: The induction of sensitization to the locomotor stimulating effects of MDMA and cocaine was independent of MDMA-induced neurotoxicity. However, the long-lasting maintenance of the sensitized response to MDMA may be related to the enduring neurotoxicity caused by MDMA. PMID- 12590355 TI - The anxiolytic CRF(1) antagonist DMP696 fails to function as a discriminative stimulus and does not substitute for chlordiazepoxide in rats. AB - RATIONALE: Compounds with a mechanism of action different from benzodiazepines may retain the anxiolytic effects of benzodiazepines with fewer side effects. CRF(1) antagonists have anxiolytic-like effects but may have different discriminative stimulus (DS) effects compared with benzodiazepines. OBJECTIVE: The present study evaluated the similarity of DS effects of a CRF(1) antagonist DMP696 to the benzodiazepine chlordiazepoxide and the ability of DMP696 to produce DS effects on its own using drug discrimination procedures, as well as its anxiolytic-like effects after acute or chronic administration. METHODS: Rats were trained to discriminate chlordiazepoxide (5.0 mg/kg, IP, 30 min prior to session) from vehicle under a fixed-ratio 10 schedule of food reinforcement and drug- or vehicle-lever selection following administration of DMP696 was determined. The effects of DMP696 on latency to exit a dark chamber (defensive withdrawal model of anxiety) were used as an index of anxiolytic-like activity. RESULTS: In chlordiazepoxide-trained rats, DMP696 (1.0-100 mg/kg, PO) resulted in most of the animals selecting the vehicle lever, as did another anxiolytic, the 5 HT(1A) partial agonist buspirone (0.3-10 mg/kg, IP). DMP696 reduced exit latency in defensive withdrawal at 10 mg/kg administered either acutely or chronically for 14 days. Thus, the doses of DMP696 studied in drug discrimination were up to 10-fold higher than those active in the anxiety model. In addition, DMP696 (10-60 mg/kg, PO) could not be established as a DS under the conditions used in this study. In a subsequent study, chlordiazepoxide was established as a DS in these same animals. CONCLUSIONS: Lack of substitution of DMP696 for the chlordiazepoxide DS in rats and its inability to acquire DS properties suggest that the DS effects of DMP696 differ from those of benzodiazepines. PMID- 12590356 TI - Changes in cognitive functioning with risperidone and olanzapine treatment: a large-scale, double-blind, randomized study. AB - OBJECTIVE: The effects of risperidone and olanzapine on cognitive functioning in patients with schizophrenia were compared in a randomized, double-blind trial. METHOD: Three hundred and seventy-seven patients were randomly assigned to receive 2-6 mg/day of risperidone or 5-20 mg/day of olanzapine for 8 weeks. Cognitive function was assessed with a focused cognitive assessment battery; in addition, extrapyramidal symptoms were assessed using the extrapyramidal symptom rating scale (ESRS), and the positive and negative syndrome scale (PANSS) was rated for all patients. RESULTS: Treatment with these two atypical antipsychotic medications was associated with improved performance on the Wisconsin card sorting test, the trail-making test, the California verbal learning test, the continuous performance test, and some aspects of verbal fluency and spatial working memory. No differences in the effects of the drugs on any of the cognitive tests were noted. Correcting for the effects of anticholinergic treatment did not alter the magnitude of cognitive effects. CONCLUSIONS: Atypical antipsychotic treatment is associated with wide-ranging benefits on cognitive functioning. Previous reports of greater benefits of olanzapine over risperidone in a small-sample pilot study were not substantiated. These results are not due in general to changes in clinical symptoms or movement disorders, suggesting a direct effect of atypical antipsychotic medications on cognitive deficits in schizophrenia. PMID- 12590357 TI - Arsenic species excretion after dimercaptopropanesulfonic acid (DMPS) treatment of an acute arsenic trioxide poisoning. AB - We studied the urinary excretion of the different arsenic species in urine samples from a young man who tried to commit suicide by ingesting about 0.6 g arsenic trioxide. He received immediate therapy with dimercaptopropanesulfonic acid (DMPS) after his delivery into the hospital. We assessed urinary arsenite (inorganic trivalent arsenic), arsenate (inorganic pentavalent arsenic), pentavalent dimethylarsinic acid (DMA) and pentavalent monomethylarsonic acid (MMA) in urine with ion-exchange chromatography and on-line hydride-technique atomic absorption spectrometry. The predominant amount of the excreted arsenic was unchanged trivalent inorganic arsenic (37.4%), followed by pentavalent inorganic arsenic (2.6%), MMA (2.1%), DMA (0.2%) and one unidentified arsenic species (0.7%, if calculated as DMA). In the first urine voiding in the clinic, the total arsenic concentration was 215 mg/l, which fell 1000-fold after 8 days of DMPS therapy. A most striking finding was the almost complete inhibition of the second methylation step in arsenic metabolism. As mechanisms for the reduced methylation efficiency, the saturation of the enzymatic process of arsenic methylation, the high dosage of antidote DMPS, which might inhibit the activity of the methyl transferases, and analytical reasons are discussed. The high dosage of DMPS is the most likely explanation. The patient left the hospital after a 12 day treatment with antidote. PMID- 12590358 TI - Cadmium-induced ectopic apoptosis in zebrafish embryos. AB - In this study, we tested the hypothesis that cadmium-induced developmental toxicity was mediated via ectopic occurrence of apoptosis during embryonic development. We employed confocal microscopy to acquire images of whole-mount staining of apoptotic cells in zebrafish embryo exposed to 100 micro M cadmium from 5 hours post fertilisation (hpf) to 28 hpf. Three-dimensional reconstruction of the images was performed and the spatial and temporal distributions of apoptotic cells in the embryos were compared. In cadmium-treated embryos with varying degrees of gross developmental malformations, significantly higher numbers of apoptotic cells were detected with this method. In order to detect the precise locations of apoptotic cells, we performed terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay in sectioned embryos. In the degenerating neural tube of cadmium-treated embryos apoptotic cells were detected, while in the healthy neural tube of the untreated controls no apoptotic cells were found. We then employed flow cytometry to investigate whether cadmium exposure would affect the dynamics of apoptosis or induce any abnormalities in cell-cycle progression. It appeared that cadmium did not induce cell-cycle arrest. The percentages of apoptotic cells did not differ in the two groups at 13, 16 or 19 hpf. At 28 hpf, however, a significantly higher percentage of apoptotic cells were found in the cadmium-treated group. Exposure to cadmium, therefore, induced ectopic apoptosis at 28 hpf without affecting the dynamics of apoptosis at earlier developmental stages. PMID- 12590359 TI - New approaches to the metabolism of xylenes: verification of the formation of phenylmercapturic acid metabolites of xylenes. AB - It was the aim of this study to ascertain whether xylenes form phenylmercapturic acids via aromatic epoxides in the human metabolism. Aromatic epoxides are suspected to exert mutagenic properties. Therefore we developed an LC/MS/MS procedure for the determination of these mercapturic acids. Using this method we were able to detect dimethylphenylmercapturic acid (DPMA) in urine samples of persons occupationally exposed to xylenes. The unequivocal LC/MS/MS detection of phenylmercapturic acid metabolites of xylene thanks to authentic standards was verified with an independent GC/MS method. Xylene concentrations in the air of the workplaces ranged between 0.7 and 58.1 ppm (median 12.6 ppm). The excretion of methylhippuric acid in the urine samples of the workers (n=27) ranges from 19.8 to 2332.5 mg/l (median 450.9 mg/l). DPMA was detected in only 9 samples of 27 exposed persons. According to a rough calculation DPMA is only formed in a ratio of 0.0003% respective to the xylene main metabolite MHA. That means that even under occupationally relevant xylene exposure potential mutagenic potencies should be negligible. DPMA in urine is not sensitive enough for general biomonitoring purposes due to the low ratio of excretion. MHA should therefore be used for biomonitoring from now on. Furthermore, our results show that irrespective of the structure of xylene isomers there is no preferences in metabolism. PMID- 12590360 TI - In vitro activation of dibromoacetonitrile to cyanide: role of xanthine oxidase. AB - Dibromoacetonitrile (DBAN) is a disinfection byproduct of chlorination of drinking water. Epidemiological studies indicate that it might present a potential hazard to human health. The present work provides evidence for DBAN activation to cyanide (CN(-)) by the hypoxanthine (HX)/xanthine oxidase (XO)/iron (Fe) system in vitro. Optimum conditions for the oxidation of DBAN to CN(-)were characterized. Addition of the sulfhydryl compounds glutathione, N-acetyl- L cysteine or dithiothreitol significantly enhanced the rate of CN(-)release. A high positive correlation existed between hydroxyl free radical ((*)OH) generation and CN(-) formation. Addition of the (*)OH scavengers mannitol or dimethylthiourea to the reaction mixtures resulted in a significant decrease in the rate of DBAN oxidation. Addition of the antioxidant enzymes catalase or superoxide dismutase resulted in a significant decrease in the rate of DBAN oxidation. The iron chelator desferrioxamine significantly decreased CN(-) formation. The maximum velocity (V(max)) and Michaelis-Menten constant (K(m)) of the reaction were assessed. Allopurinol competitively inhibited the reaction, while folic acid uncompetitively inhibited the reaction. In conclusion, (*)OH generated by the HX/XO/Fe system are implicated in DBAN oxidation. The present results represent a novel pathway for DBAN activation and might be important in explaining DBAN-induced toxicity. PMID- 12590361 TI - Comparative effect of benzanthrone and 3-bromobenzanthrone on hepatic xenobiotic metabolism and anti-oxidative defense system in guinea pigs. AB - Benzanthrone (BA) and 3-bromobenzanthrone (3-BBA) are important dye intermediates used in the manufacture of various vat and disperse dyes. BA has been implicated as a cause of hepatic malfunctions and dermal lesions in workers. However, not much information on halogenated BAs, especially 3-BBA, is available. Experiments were designed to undertake a comparative safety assessment of both BA and 3-BBA, given orally at a dose of 50 mg/kg body weight for 10 days to guinea pigs. There was a significant decrease (25%) in body weight with 3-BBA, whereas BA treatment did not cause any change. Serum glutamate oxaloacetate transaminase and glutamate pyruvate transminase were found to be significantly (P<0.05) increased in 3-BBA- as well as in BA-treated animals. 3-BBA and BA led to substantial depletion of ascorbic acid in both liver and adrenal glands. However, depletion of ascorbic acid was more pronounced with 3-BBA (19.2-28.3%) than with BA (13.5-16.6%). 3-BBA and BA treatments caused 80% and 24% depletion of hepatic free sulfydryl content, while lipid peroxidation showed a significant enhancement of 73% and 47%, respectively. BA and 3-BBA caused decreases in cytochrome P-450 content and phase I enzymes particularly ethoxyresorufin- O-deethylase and aryl hydrocarbon hydroxylase, whereas phase II enzymes (quinone reductase and glutathione- S transferase) were substantially increased. Activities of bio-antioxidant enzymes, viz., glutathione peroxidase, glutathione reductase, superoxide dismutase and catalase, were significantly increased by 153, 104, 20 and 67% in the 3-BBA treated group, whereas the degree of increase in these parameters was relatively less in BA-treated group. The data indicate that both BA and 3-BBA can disturb membrane integrity by decreasing endogenous glutathione and ascorbic acid levels with a concomitant increase in lipid peroxidative damage. This may in turn lead to impairment of hepatic P-450-dependent monooxygenase, while the changes in antioxidant enzymes reveal oxidative stress. 3-BBA treatment caused dilation of portal triad with thickening of arterial wall, hyperplasia of Kupffer cells and influx of inflammatory cells between hepatic cords, which could be due to formation of Br(*) radical or due to formation of semiquinone type of intermediate following oxidation. The results may be interpreted to mean that industrial workers exposed to 3-BBA are at higher risk than those exposed to BA, and necessary precautions should be taken to safeguard their exposure risks. PMID- 12590362 TI - Humic acid mediates iron release from ferritin and promotes lipid peroxidation in vitro: a possible mechanism for humic acid-induced cytotoxicity. AB - Humic acid (HA) has been shown to be a toxic factor for many mammalian cells, however the specific mechanism of the cytotoxicity induced by HA remains unclear. From the assessment of its redox properties, HA has been shown to be capable of reducing iron(III) to iron(II) in aqueous conditions over a broad range of pH values (from 4.0 to 9.0). By using thiobarbituric acid-reactive substances as an index, the presence of HA was noted to increase the extent of lipid peroxidation both for linoleic acids and within rat liver microsomes. In addition, the increase in HA-induced lipid peroxidation is partially inhibited by sodium azide (a singlet oxygen scavenger) or disodium 4,5-dihydroxy-1,3-benzene-disulfonic acid (a superoxide scavenger), reflecting the involvement of singlet oxygen and superoxide in the process of lipid peroxidation. The addition of HA into a reaction system has been shown to generate superoxide in a dose-dependent manner by the superoxide dismutase-inhibitable cytochrome c reduction assay. In addition, HA is able to reduce and release iron from ferritin, but this process is partially inhibited by superoxide scavengers. Subsequently, the iron released from ferritin was shown to accelerate the HA-induced lipid peroxidation. From our results we conclude that HA has the ability to reduce and release iron from ferritin storage as well as to promote lipid peroxidation. Therefore, HA coupled with released iron can disturb the redox balance and elicit oxidative stress within a biological system. This may be one of the most important mechanisms for HA-induced cytotoxicity. PMID- 12590363 TI - Changes in expression and immunolocalization of protein associated with toxic bile salts-induced apoptosis in rat hepatocytes. AB - Cholestatic liver injury results from the accumulation of toxic bile salts within the liver. The aim of the present study was to examine the temporal changes in expression and immunolocalization of protein associated with apoptosis in cholestatic rat liver. Rats were anesthetized and cholestasis was induced by double ligation of the common bile duct and sectioning between the ligatures. The animals were euthanized at day 3 and at weeks 1, 2, 4, and 6 after bile duct ligation (BDL). Apoptotic cell death was increased fivefold after 3 days of BDL, decreased over 2 weeks, and remained constant thereafter as has been demonstrated by TUNEL staining. Western blot analysis for Bax, Bcl-2, cytochrome c, and p53 were performed. Results show that total cellular Bax protein was increased 3 days after BDL and decreased over time thereafter. We observed the translocation of Bax to mitochondria and subsequent release of cytochrome c. According to our immunohistochemical data, nuclear p53 increased 3 days after BDL, but cytoplasmic sequestration of p53 was observed after 1 week. The expression of c-Myc was inhibited by 3 days, but increased at later stages following BDL. Bcl-2 was increased over time in BDL rats. Our data suggest toxic bile salts-induced hepatocellular apoptosis is related to differential expression of Bcl-2 family member protein and release of cytochrome c. Cellular localization of p53 plays an important role in apoptotic death of hepatocytes in BDL rats. PMID- 12590364 TI - Reproductive impairment in Japanese quail (Coturnix japonica) after in ovo exposure to o,p'-DDT. AB - We have previously described various effects in adult Japanese quail consequent on treatment with oestrogenic compounds in ovo. In the present study, the environmental contaminant o,p'-DDT [1-(2-chlorophenyl)-1-(4-chlorophenyl)-2,2,2 trichloroethane] was administered to quail eggs to further evaluate test endpoints for oestrogenic effects related to reproduction in the Japanese quail. Exposure to 2 mg o,p'-DDT/egg (150 micro g/g egg) resulted in impaired sexual behaviour, reduced cloacal gland area and lowered plasma testosterone concentration in males. Females displayed oviductal abnormalities, including retained right oviduct, decreased length of left oviduct, alterations in shell gland morphology and disrupted distribution of carbonic anhydrase in the shell gland. Egg laying was severely impaired. Consequently, a number of endpoints in adult quail may be useful for demonstrating an oestrogen-like mode of action by environmental contaminants during embryonic development. PMID- 12590365 TI - [Redefining schizophrenia]. PMID- 12590366 TI - [Functional neuroimaging of auditory hallucinations in schizophrenia]. AB - The neurobiological bases underlying the generation of auditory hallucinations, a distressing and paradigmatic symptom of schizophrenia, are still unknown in spite of in-depth phenomenological descriptions. This work aims to make a critical review of the latest published literature in recent years, focusing on functional neuroimaging studies (PET, SPECT, fMRI) of auditory hallucinations. Thus, the studies are classified according to whether they are sensory activation, trait and state. The two main hypotheses proposed to explain the phenomenon, external speech vs. subvocal or inner speech, are also explained. Finally, the latest unitary theory as well as the limitations the studies published are commented on. The need to continue investigating in this field, that is still underdeveloped, is posed in order to understand better the etiopathogenesis of auditory hallucinations in schizophrenia. PMID- 12590367 TI - [Evaluation of auditory hallucinations: the PSYRATS scale]. AB - INTRODUCTION: Translate and adapt the PSYRATS scale for evaluation of auditory hallucinations to Spanish and study the psychometric behavior. METHOD: After being translated into Spanish, the scale was applied to a sample of 80 patients. All had a history of auditory hallucinations, 71 with Schizophrenia and 9 with schizoaffective disorder according to DSM-IV diagnostic criteria. Demographic and clinical data were collected by a semi-structured interview, tape recording the sessions for every patient. For the reliability study, 27 patients of the sample were evaluated independently by the first four auhors of this paper. For the study of concurrent validity the results of this scale were compared with those of the Krawiecka scale in 60 patients of the sample. RESULTS: In the Spanish version, the 11 items were conserved with slight modifications in two of them. The scale showed high reliability with coefficients in the 11 items higher than 0.85. The concurrent validity showed high association between the hallucination item of the Krawiecka scale and the PSYRATS. In the factorial analysis four factors that explained 62% of the variance were obtained. CONCLUSIONS: The Spanish version of the PSYRATS (for hallucinations) is, to our knowledge, the first scale adapted to Spanish for the study of auditory hallucinations, showing excellent psychometric properties. PMID- 12590368 TI - [Diagnostic variability in a cohort of patients with multiple admissions in the last two decades]. AB - OBJECTIVE: To investigate possible changes in the admission pattern correlated with improvement in psychiatric attention and if there is variability in the diagnosis of patients previously identified as having multiple admissions during the study period. MATERIAL AND METHODS: Our study is based on fifty-nine patients who were admitted on at least ten occasions between 1983-2000. The medical records of the frequent users were reviewed and data were extracted on several variables: principal psychiatric diagnosis (DSM-IV TR criteria) and diagnostic changes as main variables and period, admissions density, admission interval and period between last admission and the end of study, as related variables. STATISTICAL ANALYSES: Friedman's variance non-parametric analysis for related samples, Pearson's chi-squared test and sequence graphing. RESULTS AND DISCUSSION: Improved psychiatric care in our city would seem to correlate with the decreasing number of patients with multiple admissions to the point where there have been no admissions in the past year. Diagnosis (DSM-IV axis I and II) variables showed important variations from one admissions to the next. The psychosocial and adaptation problems that became manifest during the successive admissions tended to improve over time as did the overall functioning of the patients. Our findings suggest that this population is composed of three distinct subgroups. Identification of the subgroups may have important implications for the management and treatment modality. PMID- 12590369 TI - [Influence of gender in vocational preferences and personality traits in Medical students]. AB - OBJECTIVE: In this paper, we try to analyze he possible relationships between gender of the Medical students, their personality and their vocational preferences. MATERIAL AND METHODS: A total of 1,482 2nd year medical students from the Valencia University were analyzed. They answered and anonymous survey on their sociodemographic characteristics and their vocational interests. We determined their personality profile with the 16 PF of Cattell. The data obtained were analyzed with the logistic regression techniques. RESULTS: Most of the population analyzed were female (62.7%) with a medium age of 20 years. The Pediatric and Gynecology-Obstetrics specialities were preferred mostly by female students and Orthopedic surgery by male students. The female students had a specific personality traits, they were more <> (A+), <> (G+), Boldness (H+), <> (n+) and <> (Q3+). While the male students were more <> (I+), <> (L+), <> (M+), <> Q2(+). CONCLUSIONS: Gender has a significant influence on the medical student both in their vocational preferences as well as their personality profile. PMID- 12590370 TI - [Venlafaxine titration dosage in depressive in-patients. A series of cases]. AB - This series of patients was gathered to assess the use of last and slow titration dosage of venlafaxine in in-patients with major depression and to evaluate the action onset. An observational open study was performed in 3 centers in 18 in patients. Nine patients were included in fast titration dosage group (FT) and 9 in slow titration dosage group (ST). As results, it was found that the FT group showed faster improvement than the ST group in HAM-D score. This improvement was sustained to the final visit, with a lower score in the FT group (6.75) than the ST group (10.67). MADRS scores up to the 15th visit were similar; however, the score was lower in the ST group on the 20th visit. Improvement was sustained to the final visit, the FT group score being lower (6.50) than the ST group score (14). CGI - i mprovement and CGI-seve rity scores we re similar to the above mentioned results. The most common events in both groups were considered mild. The data of these patients show faster response to antidepressant therapy with faster titration dosage, with maintenance of the molecule's tolerability profile. PMID- 12590371 TI - [Retrospective study of prodromal symptoms in schizophrenia]. AB - INTRODUCTION: There are some non-psychotic symptoms that can forecast the onset of psychosis. Discovering the differences between the symptoms that lead to disease and those that do not makes it possible to identify them and permits early treatment of the disease. METHODS: A sample of 689 schizophrenic patients was analyzed retrospectively. This sample was obtained from the clinical records database of the University Psychiatric Hospital Institut Pere Mata. Data were analyzed with the SPSS version 9.0 statistical package. RESULTS: The most frequent prodromal symptoms of the sample were the delusional ones, the disorganized ones and the neurotic ones. The prodromal symptoms were equally distributed in both genders. In the subtypes, paranoids showed more delusional symptoms, whereas the nonparanoids presented more disorganized symptoms. Acute onsets had more delusional prodromal symptoms whereas the insidious onsets showed more disorganized ones. CONCLUSIONS: In the prodromal stages of shizophrenia, we can also find the community neurotic prevalences regarding gender. The higher rate of neurotic symptoms in the nonparanoid subtype would be explained by the inclusion of the schizoaffective category, whereas the higher rate of disorganized symptoms categories would be due to the hebephrenic and simple categories. The latter would also explain the prodromal differences in the onset type. PMID- 12590372 TI - [New treatment for bipolar disorder in children and adolescents: learning from adult studies]. AB - BACKGROUND: Up to 45% of bipolar patients fail to respond adequately to or do not tolerate treatment with standard antimanics (lithium, valproate, carbamazepine, or olanzapine). Several new potential normothymic and antimanic treatments are under study. However, there is less literature available on new treatments for bipolar disorder in children and adolescents, but many youths with manic symptoms are treated with mood stabilizers. Our goal was to review the current literature on alternatives to lithium, valproate and carbamazepine in the treatment of bipolar disorder in children, adolescents and adults, focusing on the potential uses of these drugs in youth. METHODS: In a comprehensive computerized and manual literature search in Medline, we identified articles, abstracts and book chapters describing new treatments for bipolar disorder in children, adolescent, and adults. We also manually searched the abstracts in recent APA, AACAP and ECNP Annual Meetings. RESULTS: There are very few studies on the treatment of youths with bipolar disorder. The strongest evidence of antimanic action in this age group is on lithium, valproate, and recently on olanzapine, and adjunctive risperidone. Evidence on new antiepileptics and other novel treatments is very limited or none. CONCLUSION: More controlled studies on the treatment of children and adolescents with bipolarity are needed. Lithium, valproate, olanzapine and risperidone are probably the drugs with more evidence as antimanic efficacy in children and adolescents, but potential risks and benefits of treatment versus no treatment must be discussed with parents. PMID- 12590373 TI - [The Charles Bonnet syndrome: a case report and a brief review]. AB - The Charles Bonnet syndrome (CBS) is a rare disease that also seems to be generally misdiagnosed. Initially described in the XVIII century by the philosopher with the same name, it consist in complex visual hallucinations in elderly people who suffer sensory deprivation with no other psychopathology. The hypothesis on the neurophysiology of hallucinations suggests, as in other diseases that present hallucinations, some implication of thalamus-cortex pathway release. Some authors have proposed CBS as an early marker of dementia and Parkinson's disease. Nevertheless, the results in functional neuroimaging are not conclusive. Regarding treatment, typical and atypical neuroleptics do not seem to be useful and recent studies suggest that the new anticonvulsants could be effcient. The patient should be informed about CBS as a <>. In the case reported, the patient presented complex hallucinations with normal SPECT and neuropsychological examinations. The patient did not respond to treatment with risperidone, presenting a favorable evolucion with valpromide. Although further research is needed, this case report supports the efficacy of valpromide in CBS. PMID- 12590375 TI - [Treatment of sialorrhea in Parkinson's disease patients with clonidine. Double blind, comparative study with placebo]. AB - BACKGROUND: sialorrhea is one of the common nonmotor, non-neuropsychiatric symptoms in Parkinson's disease and its pre-sence can cause limitation in the patient's social life. The traditional treatment with anticholinergic medication is capable of triggering important neuropsychiatric complications. OBJECTIVES: this is a prospective, double-blind, placebo compared study, which follow-up Parkinson's disease patients for 3 months. METHODS: we measured the efficacy of clonidine in the management of sialorrhea. The study was performed in 32 patients (20 males and 12 females), with a mean age of 70.75 years and mean duration of the disease of 8.84 years. Randomly, 17 patients received clonidine 0.15 mg/day and the remaining 15 patients received placebo. Both groups were made up of subjects with similar characteristics, age, years of illness, sex, stage of disease (H and Y), disability (S and C) and motor score (UPDRS). Likewise, salivation affected both groups in the same intensity. We used the variable analysis and there was a p < 0.05 significance. RESULTS: the group which received clonidine showed a significant improvement of the salivation symptoms both at one month as well as at 3 months of treatment (p < 0.00001, respectively). There was no evidence of worsening of the stage of the disease, incapacity or motor score. The side effects were found only in the group that received clonidine (4 patients) without showing statistically significant. CONCLUSION: the use of clonidine can be useful in the management and treatment of sialorrhea in patients with Parkinsons disease. PMID- 12590376 TI - [Almotriptan in the treatment of migraine attacks in clinical practice: results of the TEA 2000 observational study]. AB - BACKGROUND: Almotriptan, the most recent drug of the triptan family, has shown good efficacy and tolerability profile in clinical trials. OBJECTIVE: To assess almotriptan's tolerability and effectiveness in the setting of routine clinical practice. PATIENTS AND METHODS: 1,643 patients diagnosed of migraine according to IHS criteria were recruited by 317 neurologists in the TEA 2000 study. Patients were instructed to report data on migraine attacks in a diary for a three months follow-up period. Data from 4,253 migraine attacks were obtained. RESULTS: The incidence of adverse events was 0.02 per migraine attack (3,9 % of patients). Subjective clinical improvement after 30 minutes (33.2 y 37.1 %), pain improvement after 2 hours (65.5 % and 70.2 %), pain free response after 2 hours (26.6 % and 29.2 %), recurrence between 2 and 24 hours (21.2 % and 17 %) and a complete response by 24 hours (18.6 % and 22.9 %) were found. These results were obtained in both "intention to treat" and "per protocol" analyses, being even much better when only low pain intensity attacks were considered. CONCLUSIONS: The TEA 2000 study results demonstrate good effectiveness and excellent tolerability profile of almotriptan 12.5 mg in the daily clinical neurological practice. The results of this study confirm those obtained in clinical trials carried out before almotriptan was introduced into the market and that it is a good therapeutic choice for the symptomatic treatment for migraine attacks. PMID- 12590377 TI - [Neurotrophic factors: basis for their clinical application]. AB - Neurotrophic factors are molecules that regulate neuronal survival, nervous system plasticity and many other physiological functions of neuronal and glial cells, as well as some non-neuronal tissues. They have been involved in the etiopathogenesis of some neurodegenerative disorders, and some of them have been proposed as potential treatments for these diseases on the basis of in vitro experiments and animal models. The main neurotrophic factor families with potential therapeutic applications include the family of neurotrophins (NGF, BDNF or NT-3), GDNF and related neurotrophic factor, CNTF and the members of the IGF family. Some of these molecules have already been tested in clinical trials with contradictory results. One of the major challenges to their clinical use is the difficulty to deliver them into the central nervous system. Nevertheless, solid rational exists for the possible use of neurotrophic factors in the treatment of Alzheimer's and Parkinson's diseases, peripheral neuropathies or amyotrophic lateral sclerosis. This review compiles the essential aspects of neurotrophic factors and the current studies of their clinical relevance and therapeutic potentialities. Future directions for further research are also discussed. PMID- 12590378 TI - [Hematopoietic tissue in cerebrospinal fluid?]. PMID- 12590379 TI - [Movement disorders in adult-onset measles encephalitis]. AB - There are three different neurological complications of measles infections in the brain: acute postinfectious encephalitis, acute progressive infectious encephalitis, and subacute sclerosing panencephalitis. The diagnosis of measles encephalitis (ME) is established when supported by the clinical picture, mainly of juvenile onset, and confirmed by the presence of cerebrospinal measles antibodies. Although ME is clinically characterized by progressive behavioral and mental deterioration associated with myoclonus, prior reports have suggested that adult-onset may have atypical features. We describe a 28 year-old immunocompetent man, admitted into the hospital due to a rapid motor and cognitive decline after an episode of fever and gastroenteritis. His neurological examination was significant for cognitive impairment, cervical dystonia, spontaneous and action induced myoclonus, choreiform movements, parkinsonism and ataxic gait. He was diagnosed of acute postinfectious ME based on the presence of elevated intrathecal synthesis of measles antibodies in his CSF, and a lymphocytic infiltrate of perivascular distribution without viral inclusions, with PCR negative for measles from brain biopsy. The patient continued to deteriorate to an akinetic mutism state, dying a few weeks later. Adult-onset ME is an entity rarely seen in the Western world. Although myoclonus is the most common movement disorder related to juvenile-onset ME, ataxia and other dyskinesias such as chorea, dystonia, and parkinsonism, can result from this infection when presenting in adult life. PMID- 12590380 TI - [Listeria rhombencephalitis. Neuroradiological findings]. AB - Rhombencephalitis due to listeria monocytogenes is an uncommon and serious form of brainstem infection. The disease has a characteristic biphasic course: a nonspecific prodrome of headache, nausea or vomiting, and fever lasting for a several days is followed by progressive asymmetrical cranial-nerve palsies. We report two cases of Listeria monocytogenes rhombencephalitis. The first case is a previously healthy 20 year-old-man who developed fever, headache, nausea and vomiting, followed by numbness in left trigeminal nerve. The second case is an immunosuppressed 77 year-old-man, who developed sudden left hemiparesis, followed by fever and severe brainstem dysfunction with ophthalmoplegia and dysphagia. In both cases, a brain magnetic resonance imaging (MRI) scan, showed increased intensity on T2-weighted lesions in the brainstem that enhanced after contrast on T1-weighted sequences. Both patients had a favorable outcome with full clinical recovery. We conclude that MRI aids in the early detection of parenchymal infections, therefore, MRI is crucial for early diagnosis and is very useful for follow-up examinations. PMID- 12590381 TI - [Posterior fossa venous epidural hematoma. Based on 2 cases]. AB - We present two rare cases of posterior fossa epidural hematoma (PFEH) due to occipital fracture and tearing of transverse sinus. One of the patients showed supratentorial and infratentorial extension, which is found in half of the PFEH cases. Both cases experienced clinical worsening within 48 h of the head injury, with control CT showing increase of haematoma volume. Both patients were elected for surgery consisting of suboccipital craniectomy, control of sinus bleeding and sealing of epidural space with dural fixation. Sources of bleeding of PFEH are, by order of frequency, transverse sinus, small dural arteries and sigmoid sinus. PFEH may cause fatal clinical outcome with less volume than supratentorial ones. PFEH have high mortality if not treated. Patients with PFEH and GCS <15 should be admitted to hospital and a control CT should be done if there is deterioration or in the first 24 hours. The treatment of choice is surgery, conservative therapy being an alternative for small PFEH and good clinical evolution. PMID- 12590382 TI - [Prolonged clinical pattern of brain death in patients under barbiturate sedation]. PMID- 12590383 TI - Epsilon -N,N,N-trimethyllysine-specific ions in matrix-assisted laser desorption/ionization-tandem mass spectrometry. AB - Epsilon -N,N,N-trimethyllysine (K(me3)) is a component of a number of proteins and plays an important role in the expression of their biological functions. Trimethylation, which causes an incremental increase in mass of 42.0470 Da from that of the corresponding MH(+) ion, cannot be distinguished from the acetylation (+42.0106 Da), which also occurs on epsilon-amino groups of Lys or alpha-amino groups in many proteins, without high-accuracy mass measurement which is accurate to within the second decimal place. MALDI-MS and MS/MS have been applied for the analyses of post-translational modifications of histone H3, which is known to contain both multiple acetylation and methylation sites in its sequence. During the measurements of the modified peptides, a novel fragmentation which involves the loss of trimethylamine from K(me3) was found. This characteristic fragmentation, which was observed to produce ions separated by 59 Da from the conventional precursor ion or sequence ions, would be useful for probing K(me3) units in the sequence. PMID- 12590384 TI - Mass spectrometric investigation of N-sulfonylated purine nucleic bases and nucleosides. AB - The gas/phase behaviour of N-sulfonylated purine nucleic bases and nucleosides towards electron impact (EI) and matrix-assisted laser desorption/ionization (MALDI) occurring in a ion trap of a Fourier transform ion cyclotron resonance mass spectrometer is investigated. The influence of the storage time on the protonated molecule ([M+H](+)) abundance under EI conditions confirms that the formation of these ions proceeds through ion/molecule reactions. Using stored waveform inverse Fourier transform (SWIFT) selective isolation of M(+.) or H(3)O(+), self-chemical ionization, M(+.)/M, and chemical ionization, H(3)O(+)/M, are detected. Investigation of specific EI expulsion of SO(2), SO(2)H and/or SO(2)H(2) from M(+.) and/or [M+H](+) shows that oxygen protonation in bond;SO(2)bond; proceeds faster than nitrogen protonation. Expulsion of SO(2) from molecular ions is not observed in MALDI mass spectra of nucleosides. PMID- 12590385 TI - Determination of perfluorooctane sulfonate in river water by liquid chromatography/atmospheric pressure photoionization mass spectrometry by automated on-line extraction using turbulent flow chromatography. AB - A simple, fast and sensitive liquid chromatography/atmospheric pressure photoionization mass spectrometry (LC/APPI-MS) method, with automated on-line extraction using turbulent flow chromatography (TFC), was developed for the determination of perfluorooctane sulfonate (PFOS) in river water. In this method, following an on-line extraction by injection onto a column under TFC conditions, PFOS is back-flushed onto a reversed-phase column via on-line column switching, and resolved chromatographically at a laminar flow rate of 1 mL min(-1). Using this tandem LC-LC/APPI-MS system the extraction, separation and selective detection of PFOS in river water could be achieved with satisfactory selectivity and sensitivity. The limit of detection (LOD) (S/N = 3) and the limit of quantitation (LOQ) (S/N = 10)were 5.35 and 17.86 pg mL(-1). The described procedure was very simple since no off-line sample preparation was required, total analysis time being 18.75 min. PMID- 12590386 TI - A novel nanoflow interface for atmospheric pressure ionization mass spectrometry. AB - A novel spray-ionization technique for nanoflow liquid chromatography/mass spectrometry (nLC/MS) has been developed by modifying the sonic spray ionization (SSI) technique. A solution from a tapered fused-silica capillary is sprayed by a gas flow coaxial to the capillary, and ions produced are analyzed with an ion trap mass spectrometer. The ion intensity is shown to have a steep threshold at a low gas velocity and to be much less dependent on the gas velocity than that of conventional SSI, in which the ion intensity is strongly dependent on the gas velocity and reaches its maximum at sonic velocity. Thus, we conclude that the concentration of charge in the solution at the tapered capillary tip with an inner diameter of 15 microm is almost at saturation so that charged droplets are produced from the solution by electrical force, rather than by sheer stress due to the gas flow. The ions are readily produced from these charged droplets. Preliminary results are compared with results obtained with a miniaturized electrospray unit. PMID- 12590387 TI - Partial pressure dependency of 17O/16O and 18O/16O of molecular oxygen in the mass spectrometer. AB - A method to determine both (17)O/(16)O and (18)O/(16)O ratios for molecular oxygen with high precision by direct introduction into the mass spectrometer without gas separation is presented. Because both (17)O/(16)O and (18)O/(16)O in mixed gases have good linear correlations with their mixing ratios, these isotopic compositions can be determined without a gas-separation procedure by calibration using prepared standard gases with variable mixed ratios and by monitoring the amounts of fragment ions. Analytical precision for delta(17)O and delta(18)O of 45 and 7 per meg, respectively, were obtained. The observed partial pressure dependency of isotopic composition may be caused by isotope fractionation during admission from the ionization chamber into the flight tube of the mass spectrometer. PMID- 12590388 TI - Analysis of rabbit tear proteins by high-pressure liquid chromatography/electrospray ionization mass spectrometry. AB - The aim of this study was to develop a fast and reliable analytical procedure for the display of the protein components of tears that can be used to differentiate the status of the ocular surface. Using this new procedure, we analyzed the tear protein components following a corneal wound in the rabbit. Calibrated 10-microL glass, fire-polished capillary micropipettes were used to collect tears from New Zealand White rabbits prior to and daily for 9 days following a unilateral 6-mm diameter centrally placed anterior keratectomy. Tear proteins were eluted by a reversed-phase high-performance liquid chromatography (RP-HPLC) column and the tear protein profile was monitored by electrospray ionization (ESI) mass spectrometry positive total ion current (TIC) chromatography. Tear proteins were reliably separated into 17 peaks, each of which contained one or a number of protein components. The molecular weight of each protein component was determined by on-line ESI. Major tear protein components, lactoferrin, lysozyme (minimally detectable in rabbit tears), albumin, lipocalin, lipophilin and beta2 microglobulin, were tentatively identified by this method. Based on the mass spectrometric data, beta2-microglobulin was found to be glycosylated with N acetylhexosamine. ESI-positive TIC chromatograms and mass spectra revealed comparative differences in the tear protein spectra after corneal wounding. One day after wounding, rabbit lysozyme with a molecular weight of 14,717 Da was found to be 8-fold higher in the tears of wounded eyes when compared with tears from unwounded eyes. It dropped back to normal 3 days after wounding. The expression of an unidentified tear protein with the molecular weight of 16,060 Da was also elevated after corneal wounding and returned to normal level by day 5. In this study, LC/ESI-MS was developed as a fast, reproducible and simple method for the identification and analysis of many of the protein components of the tears. Importantly, this technique also allows quantification of each component resolved in the chromatogram. This method is very suitable for mapping peptides and proteins (<80 kDa) in tears. PMID- 12590389 TI - Individual variability in Tityus serrulatus (Scorpiones, Buthidae) venom elicited by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Venom variability in specimens of Tityus serrulatus scorpion was assessed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) analyses. An expanded time lag venom extraction protocol was carried out using ten scorpions to study individual variations that might occur due to different rates in protein expression and/or processing. The first extraction of venom was made from the animals after 20 days of starvation, which allowed the venom gland to be filled up. The second extraction event was carried out 24 hours after the first one. The third was 8 days after the first extraction. By means of MALDI-TOF analyses, important variations were observed in venoms of a single specimen extracted at different times, especially in latter extraction events. These variations are most probably related to dynamics in cell gland production. Since T. serrulatus is a parthenogenetic species, sexual variations are naturally excluded and we did not expect intra-specific variations, which was confirmed. Knowledge of individual venom variability is extremely important to avoid misunderstandings in the use of venom proteomic analysis as a taxonomic tool. PMID- 12590390 TI - Automated quantitative and isotopic (13C) analysis of dissolved inorganic carbon and dissolved organic carbon in continuous-flow using a total organic carbon analyser. AB - A method for the automated (13)C analysis of dissolved inorganic and organic carbon species has been developed to operate on a continuous-flow isotope ratio mass spectrometer (CF-IRMS). For natural and anthropogenic carbon species, the (13)C stable isotope has proven to be an excellent environmental tracer. Analytical performance tests were carried out on various organic compounds from easily oxidisable (sugar) to difficult (humic acid). A set of natural samples was also analysed to confirm the flexibility of the system. Analytical precision (2sigma) is typically <0.20 per thousand with sample reproducibility from 0.10 0.35 per thousand depending on reactivity of material. We believe this to be the first successful use of a total organic carbon (TOC) analyser for both dissolved inorganic and, specifically, dissolved organic species for (13)C stable isotope analysis in an automated CF-IRMS system. Routine analysis is achieved fairly quickly, is relatively simple with little or no sample manipulation, and will allow new and exciting studies for stable isotope research in both natural abundance and organic tracer studies not easily achieved before. PMID- 12590391 TI - Heuristic charge assignment for deconvolution of electrospray ionization mass spectra. AB - We propose a new algorithm for deconvolution of electrospray ionization mass spectra based on direct assignment of charge to the measured signal at each mass to-charge ratio (m/z). We investigate two heuristics for charge assignment: the entropy-based heuristic is adapted from a deconvolution algorithm by Reinhold and Reinhold;10 the multiplicative-correlation heuristic is adapted from the multiplicative-correlation deconvolution algorithm of Hagen and Monnig.6 The entropy-based heuristic is insensitive to overestimates of z(max), the maximum ion charge. We test the deconvolution algorithm on two single-component samples: the measured spectrum of human beta-endorphin has two prominent and one very weak line whereas myoglobin has a well-developed quasi-gaussian envelope of 17 peaks. In both cases, the deconvolution algorithm gives a clean deconvoluted spectrum with one dominant peak and very few artefacts. The relative heights of the peaks due to the parent molecules in the deconvoluted spectrum of a mixture of two peptides, which are expected to ionize with equal efficiency, give an accurate measure of their relative concentration in the sample. PMID- 12590392 TI - A simple method for fabrication of silver-coated sheathless electrospray emitters. AB - A simple and time-saving procedure is proposed for preparing a silver-coated fused-silica capillary tip. The tapered capillary tip was coated with silver using an acrylic-based silver conductive pen. The fabrication of a silver-coated fused-silica capillary tip takes less than 5 minutes. The silver-coated fused silica capillary tip is rugged and durable for sheathless capillary electrophoresis/electrospray mass spectrometry and also for microspray applications. PMID- 12590393 TI - Structural studies of glutenin subunits 1Dy10 and 1Dy12 by matrix-assisted laser desorption/ionisation mass spectrometry and high-performance liquid chromatography/electrospray ionisation mass spectrometry. AB - Structural studies of the high molecular weight (HMW) glutenin subunits 1Dy10 and 1Dy12 of bread wheat were conducted using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS) and reversed phase high-performance liquid chromatography/electrospray ionisation mass spectrometry (RP-HPLC/ESI-MS). For both proteins, MALDI-TOFMS analysis showed that the isolated fractions contained a second component with a mass about 500 540 Da lower than the major component. The testing and correction of the gene derived amino acid sequences of both proteins were performed by direct MALDI TOFMS analysis of their tryptic peptide mixture and analysis of the digests was performed by recording several MALDI mass spectra of the mixture at low, medium and high mass ranges, optimising the matrix and the acquisition parameters for each mass range. Complementary data were obtained by RP-HPLC/ESI-MS analysis of the tryptic digest. This resulted in the coverage of the whole protein sequences except for two short fragments (T1 and T8), which are identical in the two homologous subunits, and for an additional dipeptide (T14) in subunit 1Dy12, which were not detected. It also demonstrated that, in contrast to the gene derived data, the sequence of subunit 1Dy12 does not include the dipeptide Gly Gln between residues Gln(454) and Pro(455), and that the lower mass components present in both fractions correspond to the same sequences lacking short peptides that are probably lost from the protein N- or C-termini. Finally, the results obtained provide evidence for the lack of a substantial level of glycosylation or other post-translational modifications of the two subunits, and demonstrate that mass spectrometric mapping is the most useful method presently available for the direct verification of the gene-derived sequences of HMW glutenin subunits and similar proteins. PMID- 12590394 TI - High affinity capture surface for matrix-assisted laser desorption/ionisation compatible protein microarrays. AB - A surface for the capture of biotin-tagged proteins on matrix-assisted laser desorption/ionisation (MALDI) targets has been investigated. Binding of a poly-L lysine poly(ethylene glycol)-biotin polymer to glass and gold surfaces has been demonstrated using dual wavelength interferometry. Biotinylated proteins were captured onto this surface using tetrameric neutravidin as a multivalent bridging molecule. Biotin tagging of proteins was achieved by chemical biotinylation or by expressing a protein with a biotinylation consensus sequence in E. coli. The specificity of the surface for biotin-tagged proteins allowed the purification of biotin-tagged glutathione-S-transferase from a bacterial lysate directly onto a MALDI target. Subsequently, the protein was digested on the MALDI target and a protein fingerprint analysis confirmed its presence directly, but no E. coli proteins were detected. Therefore, we conclude that this surface is highly specific for the capture of biotin-labelled proteins and has low non-specific binding properties for non-biotinylated proteins. Furthermore, protein-protein interactions using biotinylated lectins were investigated, and the selective capture of the glycoprotein fetuin with wheat germ agglutinin was demonstrated. Also, immobilised Arachis hypogea agglutinin recognised a minor asialo component of this glycoprotein on the array. The high affinity immobilisation of proteins onto this surface allowed effective desalting procedures to be used which improved the desorption of high molecular weight proteins. Another aspect of this surface is that a highly ordered coupling of the analyte can be achieved which eliminates the search for the sweet spot and allows the creation of densely packed protein microarrays for use in mass spectrometry. PMID- 12590395 TI - Validating methods for measuring delta18O and delta13C in otoliths from freshwater fish. AB - The ability of the phosphoric acid digestion technique to extract carbon dioxide from biogenic carbonates and reliably reproduce delta(18)O and delta(13)C signatures from standard reference materials (NBS-18, NBS-19) was tested and shown to produce accurate, unbiased measurements of non-biologic materials. The effects of roasting preparation methods commonly reported when analyzing biogenic carbonates were also tested in a series of experiments using reference standards and otoliths obtained from aquacultured Arctic charr and rainbow trout. Roasting had no effect on the isotope measurement of reference standards. No significant differences between mean oxygen isotope signatures from paired experiments with roasted and non-roasted fish otoliths were found. However, otolith oxygen isotope measurements were significantly enriched in comparison to rearing water-based measurements for both species. Agreement between expected isotopic equilibrium and measured otolith delta(18)O values varied as a function of roasting temperature and between species. Criteria for the selection of appropriate roasting temperatures are suggested and favour 350 degrees C in freshwater fish where unbiased estimates of average rearing water temperatures and known differences in rearing temperatures were obtained. Carbon isotopic disequilibria were observed for both species. A mixing model analysis established differences in the percentage of metabolically derived carbon in studied otoliths, with Arctic charr deriving a greater proportion of otolith delta(13)C from metabolism as a result of higher metabolic rates. PMID- 12590396 TI - Application of tandem mass spectrometry combined with gas chromatography and headspace solid-phase dynamic extraction for the determination of drugs of abuse in hair samples. AB - A new method combination, headspace solid-phase dynamic extraction coupled with gas chromatography/tandem mass spectrometry (HS-SPDE/GC/MS/MS), is introduced to determine drugs of abuse in hair samples. This highly automated procedure utilizes SPDE for pre-concentration and on-coating derivatization as well as GC and triple quadrupole MS/MS for selective and sensitive detection. All these steps, apart from washing and cutting of the hair samples, are performed without manual intervention on a robot-like autosampler.SPDE is a solventless extraction technique related to solid-phase microextraction (SPME). The analytes are absorbed from the sample headspace directly into a hollow needle with an internal coating of polydimethylsiloxane by repeated aspirate/dispense cycles.The HS SPDE/GC/MS/MS procedure was applied to the analysis of methadone, the trimethylsilyl derivatives of cannabinoids and the trifluoroacetyl derivatives of amphetamines and designer drugs. The method was shown to be sensitive with detection limits between 6 and 52 pg/mg hair matrix and precision between 0.4 and 7.8% by the use of an internal standard technique. Linearity was obtained from 0.1-20 ng/mg with coefficients of correlation between 0.995 and 0.999. Compared with conventional methods of hair analysis, HS-SPDE/GC/MS/MS is easier to use, substantially faster, with the degree of sensitivity and reproducibility demanded in clinical and forensic toxicology. The main advantage of the SPDE technique in relation to SPME is the robustness of the capillary. PMID- 12590397 TI - Analysis of protein profiles of genetically modified potato tubers by matrix assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Traceability of genetically modified (GM) foods demands the development of appropriate reliable techniques in order to identify and quantify peptide or nucleic acid residues in GM plants and food products through the food chain. In this study the applicability of matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOFMS) was demonstrated for the characterization of proteins of transformed and untransformed potato (Solanum Tuberosum L.) tubers. In GM tubers the expression level of the G1-1 gene, which regulates transition from dormancy to sprouting tubers, was inhibited by antisense technology. The analysis of antisense transformed lines showed that several of them exhibited a significant delay in sprouting relative to the control lines, in accordance with a decrease in the transcript level. Preliminary attempts to compare the protein patterns obtained from transformed and control lines using traditional electrophoresis were not able to reveal differences in the low-kDa range. Instead, MALDI-TOFMS applied to total peptide extract without any purification was able to distinguish spectral patterns of transformed and untransformed lines. In particular, several characteristic peaks from m/z 4373 to 4932 were detected only in the mass spectra of GM tuber samples. PMID- 12590399 TI - Fragmentation mechanisms in electron ionization mass spectrometry of some 2-alkyl and 2-aryl-4-arylidene-5(4H)-oxazolones. PMID- 12590398 TI - Use of S-pentafluorophenyl tris(2,4,6-trimethoxyphenyl)phosphonium acetate bromide and (4-hydrazino-4-oxobutyl) [tris(2,4,6-trimethoxyphenyl)phosphonium bromide for the derivatization of alcohols, aldehydes and ketones for detection by liquid chromatography/electrospray mass spectrometry. AB - The synthesis of S-pentafluorophenyl tris(2,4,6-trimethoxyphenyl)phosphonium acetate bromide (TMPP-AcPFP) and the novel compound (4-hydrazino-4-oxobutyl) [tris(2,4,6-trimethoxyphenyl)phosphonium bromide (TMPP-PrG) is described and the use of these compounds as derivatizing reagents for alcohols, aldehydes and ketones evaluated. Methods have been developed for the pre-column derivatization of alcohols using TMPP-AcPFP and for aldehydes and ketones using TMPP-PrG. The reactions were investigated by the use of a variety of individual test compounds containing the target functional groups. The TMPP acetyl ester and TMPP propyl hydrazone derivatives formed with their respective target analytes produced an enhanced response in electrospray ionization mass spectrometry (ESI-MS), and reproducible chromatography. The use of these two reagents to derivatize and facilitate detection of alcohols (including sugars and steroids), aldehydes and ketones (including steroids) by LC/ESI-MS was investigated. PMID- 12590400 TI - Unimolecular metastable decompositions of 1,1,1-trifluoroisopropyl methyl ether [CF3(CH3)CHOCH3] upon electron ionization. PMID- 12590401 TI - Cognitive deficits and cognitive normality in recreational cannabis and Ecstasy/MDMA users. PMID- 12590402 TI - Mirtazapine in combination with amitriptyline: a drug-drug interaction study in healthy subjects. AB - OBJECTIVE: To assess the steady-state pharmacokinetics of mirtazapine (30 mg/day orally) and amitriptyline (75 mg/day orally) during combined administration compared with that of either drug administered alone. To evaluate the tolerability and effects on psychometric tests of acute and subchronic administration of both drugs combined and alone. METHODS: In a single-blind, three-way cross-over study, 24 (12 male and 12 female) healthy subjects were randomly assigned to six different sequences of three 9-day treatments, i.e. racemic mirtazapine (30 mg/day), amitriptyline (75 mg/day) or the combination of these drugs. To control for acute pharmacodynamic assessments, during the first treatment period, a placebo group (n = 8; 4 females and 4 males) was added. Serial blood samples were drawn for plasma level measurements that were subsequently subjected to pharmacokinetic analysis. Psychometric tests assessed attentional performance, and a computer-assisted telephone questionnaire assessed self-ratings of drowsiness/alertness and sleep quality. RESULTS: Amitriptyline increased the C(max) of mirtazapine (+ 36%, p < 0.05) in male subjects only. Mirtazapine altered the C(max) of amitriptyline in both male (+ 23%, p < 0.05) and female (- 23%, p < 0.05) subjects. No changes were observed for other pharmacokinetic parameters. Metabolite parameters were not affected. Changes in parent compound levels mainly resulted from effects on absorption. The psychometric test results did not reveal significant changes between combined and single drug treatments. The telephone registrations of VAMRS and LSEQ did not show clinically relevant differences between the active treatments. CONCLUSION: Combined administration of mirtazapine (30 mg/day) and amitriptyline (75 mg/day) alters the pharmacokinetics of either compound to a minor extent. Adding one drug to the other and substituting one drug by the other had no major effects on tolerability. Nevertheless, caution is warranted when combining amitriptyline and mirtazapine. PMID- 12590403 TI - Seasonal variations in the binding of [3H]paroxetine to the platelet serotonin transporter sites in African-American cocaine-dependent patients and healthy volunteers. AB - Although seasonal fluctuations in several indices of serotonin function have been described, little is known about seasonality and serotonergic activity in substance abusers. We investigated whether there were seasonal differences in platelet serotonin transporter sites among cocaine dependent patients and controls. Platelet [(3)H]paroxetine binding, a measure of serotonin transporter sites, was assayed in 141 African-American cocaine-dependent subjects and in 60 race matched healthy volunteers who served as the control group. B(max) (density of serotonin transporter) and K(d) (affinity constant) values of [(3)H]paroxetine binding were compared during spring, summer, fall and winter. Consistent with our previous findings B(max) values were significantly lower in cocaine patients (639 +/- 234) than in controls (906 +/- 225) (t = 7.12, p < 0.001). Moreover, B(max) values showed a significant seasonal variation in controls with the highest values in summer and spring compared with fall and winter (F = 4.47, p < 0.01). However, there were no significant seasonal differences in B(max) values in cocaine patients. K(d) values did not show any seasonal changes in either group. There were no effects of age or gender on seasonal variations in B(max). The study demonstrates a seasonal effect on platelet serotonin uptake in healthy African-American volunteers. The lack of seasonal differences in transporter availability in cocaine patients indicates that the normal seasonal rhythm of serotonergic activity may be disturbed in cocaine abusers. Biological studies that employ platelet serotonin transporter sites as a marker of serotonin function should consider seasonal variations in these markers as a potential source of variance. PMID- 12590404 TI - Fluvoxamine as effective as clomipramine against symptoms of severe depression: results from a multicentre, double-blind study. AB - BACKGROUND: Although selective serotonin reuptake inhibitors (SSRIs) are better tolerated than tricyclic antidepressants, their efficacy in severe depression remains to be further elucidated. METHOD: A double-blind, multicentre study was conducted in 86 severely depressed inpatients (>or= 25 on the 17-item Hamilton depression rating scale [HAMD] total score) to compare the efficacy and safety of fluvoxamine with that of clomipramine. Following placebo run-in, 86 patients were randomised to receive fluvoxamine or clomipramine (100-250 mg/day) for 8 weeks. RESULTS: Fluvoxamine and clomipramine both resulted in marked improvements; there were no statistically significant differences between them on the 17-item HAMD total score, the clinical global impression severity of illness or global improvement items or the Montgomery-Asberg depression rating scale, at any visit. At the end of the study, 71% in the fluvoxamine group and 69% in the clomipramine group were responders (>or= 50% decrease in 17-item HAMD total score). However, fluvoxamine was better tolerated than clomipramine. Clomipramine was associated with a higher incidence of overall and treatment-related adverse events. In addition, the percentage of patients discontinued prematurely due to adverse events was more than twice as high with clomipramine than with fluvoxamine (24% vs 11%). CONCLUSION: Fluvoxamine and clomipramine are equally effective in severe depression, but fluvoxamine has a better safety and tolerability profile. PMID- 12590405 TI - Effect of citalopram on ouabain-induced arrhythmia in isolated guinea-pig atria. AB - The effect of citalopram (CTP) a selective serotonin reuptake inhibitor agent was studied on ouabain-induced arrhythmia in spontaneously beating isolated guinea pig atria. CTP (2-32 microg/ml) produced a dose-dependent decrease in the force of contractions (7-62%), and in the rate of contractions (11-72%). Pre administration of the atria with CTP inhibited the ouabain-induced arrhythmia in isolated atria. Ouabain alone (1.2 microg/ml) produced arrhythmia at 4.5 min, and asystole at 20.7 min. Pretreatment with CTP (8 microg/ml) significantly increased the time of onset of arrhythmia to 9.5 min. In addition CTP prolonged the beating of atria (survival time) to more than 56 min, and inhibited the occurrence of asystole. These findings indicate that CTP produces direct cardiac action, probably due to the inhibition of cardiac Na(+) and Ca(2+) channels. Moreover our results suggest that CTP may reduce the membrane conductance through inhibition of ionic channels which decrease ouabain-induced arrhythmia. PMID- 12590406 TI - A preliminary study of baclofen-induced growth hormone release in generalised social phobia. AB - OBJECTIVE: The authors sought to examine central GABA(B) receptor responses in patients with generalised social phobia using the growth hormone (GH) response to baclofen. METHOD: Baclofen 20 mg was administered to 15 patients and 15 healthy matched controls and plasma growth hormone was monitored over a 3 h period. RESULTS: The GH response in patients was significantly reduced in comparison to that of controls when calculated as both area under the curve (AUC) and delta (Delta) response. CONCLUSIONS: The results suggest an abnormality of central GABA(B) receptor function in this disorder. PMID- 12590407 TI - Low doses of alcohol have a selective effect on the recognition of happy facial expressions. AB - Alcohol is one of the most widely used recreational drugs, yet it is associated with undesirable social behaviour. It is used primarily for its psychoactive properties, increasing sociability and talkativeness. We hypothesize that low doses of alcohol can improve the performance related to positive emotional cognition. In this experiment, we examined the effect of low doses of alcohol on the processing of emotional facial expressions. Fifteen young male volunteers drank alcohol at volumes of 30, 60, 120 ml (0.14, 0.28, 0.56 g/kg) and performed discrimination tasks on morphed facial emotion expressions of anger, happiness, sadness and surprise-neutral. One-way ANOVA co-varying pretreatment performances revealed significant differences between alcohol levels in happy face discrimination ( p<0.01). Bonferroni correction demonstrated that low doses of alcohol caused a significantly better discrimination of happy faces, and that the performances were worse with higher doses ( p<0.001). No significance was observed with the other three emotional faces. These results indicate that low doses of alcohol affect positive emotional cognition of happy facial expressions. PMID- 12590408 TI - Coffee increases state anxiety in males but not in females. AB - Coffee, reproducing the conditions under which caffeine is normally ingested, containing 3, 75, 150 or 300 mg of caffeine was given to healthy male and female volunteers. 25-30 min after drinking the beverage, they completed the Spanish version of the state-trait anxiety inventory (STAI). The beverage increased state anxiety, in a dose-dependent manner, in males but not in females. This could be due to a lesser sensitivity of females to coffee. PMID- 12590409 TI - Inter--rater and test--retest reliability of the Japanese version of the subjective deficit syndrome scale. AB - The subjective deficit syndrome scale (SDSS) was established in 1990 by Jaeger et al. to assess systematically subjective experiences of schizophrenic patients. It was translated into Japanese in 1999 by this paper's second author (T.I.). In this study, the inter-rater reliability of this Japanese version (SDSS-J) was examined. The subjects studied were 13 schizophrenic patients (male 6, female 7; respective average ages at study entry and disease onset: 41 and 25 years), who were being followed up at the outpatients' service of the Department of Neuropsychiatry, Hyogo College of Medicine. Four psychiatrists attended together, the systematic interview with each subject to rate the SDSS-J and independently rated 19 items. The severity scales of the items assessed by these raters ranged from 0 to 4 for 12 items and from 0 to 3 for 7. The ANOVA ICC inter-rater reliability values for the 19 individual items ranged from 0.67 to 0.96. The ANOVA ICC test-retest reliability values achieved by two raters were also high overall, ranging from 0.72 to 1.00, except for one item (item 13) assessed by one rater. Our results suggest that the SDSS-J is a potentially useful rating scale for evaluating subjective experiences of schizophrenic subjects. PMID- 12590410 TI - A comparison of group sequential methods for binary longitudinal data. AB - Interim analyses are conducted to allow for early termination of the trial, for ethical as well as economical reasons. Here we consider interim analyses in repeated measurements studies where the measurements are binary. Two methods for analysing this kind of data are compared according to their operating characteristics. A subject-specific approach based on the logistic random-effects model is compared with the population-averaged approach based on the generalized estimating equations. The comparison is illustrated with simulations using a randomized clinical trial for toenail fungal infection. PMID- 12590411 TI - Missing data in the 2 x 2 table: patterns and likelihood-based analysis for cross sectional studies with supplemental sampling. AB - Standard measures of crude association in the context of a cross-sectional study are the risk difference, relative risk and odds ratio as derived from a 2x 2 table. Most such studies are subject to missing data on disease, exposure, or both, introducing bias into the usual complete-case analysis. We describe several scenarios distinguished by the manner in which missing data arise, and for each we adjust the natural multinomial likelihood to properly account for missing data. The situations presented allow for increasing levels of generality with regard to the missing data mechanism. The final case, quite conceivable in epidemiologic studies, assumes that the probability of missing exposure depends on true exposure and disease status, as well as upon whether disease status is missing (and conversely for the probability of missing disease information). When parameters relating to the missing data process are inestimable without strong assumptions, we propose maximum likelihood analysis subsequent to collecting supplemental data in the spirit of a validation study. Analytical results give insight into the bias inherent in complete-case analysis for each scenario, and numerical results illustrate the performance of likelihood-based point and interval estimates in the most general case. Adjustment for potential confounders via stratified analysis is also discussed. PMID- 12590412 TI - Estimating the probability of toxicity at the target dose following an up-and down design. AB - One of the most important aspects of a phase I trial or other acute toxicity study is estimating accurately the probability of toxicity that is associated with the recommended dose. We use the biased coin up-and-down design to allocate and isotonic regression to estimate toxicity probabilities and determine the recommended dose. We then derive, using bootstrap methods, an estimate of the probability of toxicity at the recommended dose. Small sample properties of this estimator are also evaluated. Published in 2003 by John Wiley & Sons, Ltd. PMID- 12590413 TI - Bias due to missing exposure data using complete-case analysis in the proportional hazards regression model. AB - We studied bias due to missing exposure data in the proportional hazards regression model when using complete-case analysis (CCA). Eleven missing data scenarios were considered: one with missing completely at random (MCAR), four missing at random (MAR), and six non-ignorable missingness scenarios, with a variety of hazard ratios, censoring fractions, missingness fractions and sample sizes. When missingness was MCAR or dependent only on the exposure, there was negligible bias (2-3 per cent) that was similar to the difference between the estimate in the full data set with no missing data and the true parameter. In contrast, substantial bias occurred when missingness was dependent on outcome or both outcome and exposure. For models with hazard ratio of 3.5, a sample size of 400, 20 per cent censoring and 40 per cent missing data, the relative bias for the hazard ratio ranged between 7 per cent and 64 per cent. We observed important differences in the direction and magnitude of biases under the various missing data mechanisms. For example, in scenarios where missingness was associated with longer or shorter follow-up, the biases were notably different, although both mechanisms are MAR. The hazard ratio was underestimated (with larger bias) when missingness was associated with longer follow-up and overestimated (with smaller bias) when associated with shorter follow-up. If it is known that missingness is associated with a less frequently observed outcome or with both the outcome and exposure, CCA may result in an invalid inference and other methods for handling missing data should be considered. PMID- 12590414 TI - Methods for categorizing a prognostic variable in a multivariable setting. AB - The literature is filled with examples of categorization of a continuous prognostic variable in a univariable setting followed by the addition of this categorical variable to an existing multivariable model. Typically, an "optimal" cutpoint for a new prognostic variable is obtained through a systematic search relating the variable to the outcome in an univariable manner. The corresponding categorical variable is then fitted in a multivariable model along with other already established prognostic covariates to assess the additional value of the new variable. This prompts the question whether the cutpoint search should have been performed in the same multivariable setting where it will ultimately be used. In this paper, we extend the univariable cutpoint search methods (split sample approach and two-fold cross-validation approach) to the multivariable setting using -2 x log-likelihood statistic as the correlative measure. A Monte Carlo simulation study demonstrates that both methods are more efficient in detecting the true cutpoint and in estimating the effect size under the multivariable setting as opposed to the univariable setting. The cross-validation method performs better than the split-sample method in univariable as well as multivariable scenarios. For the cross-validation method in the multivariable setting, there is still a substantial loss of power when a cutpoint model is used in cases where there is a continuous relationship between the covariate and the outcome. An example is provided to illustrate the value of the multivariable cross-validation approach. PMID- 12590415 TI - Comparison of HIV-1 and HIV-2 infectivity from a prospective cohort study in Senegal. AB - From a prospective cohort study of 1948 initially human immunodeficiency virus (HIV) uninfected female commercial sex workers followed between 1985 and 1999 in Dakar, Senegal, the authors compared the male to female per infectious sexual exposure transmission probability of HIV types one (HIV-1) and two (HIV-2). New non-parametric competing risks failure time methods were used, which minimized modelling assumptions and controlled for risk factors for HIV infection. The HIV 1 versus HIV-2 infectivity ratio over time was estimated by the ratio of smoothed non-parametric kernel estimates of the HIV-1 and HIV-2 infection hazard functions in sex workers, adjusted by an estimate of the relative HIV-1 versus HIV-2 prevalence in the partner population. HIV-1 was found to be significantly more infectious than HIV-2 throughout the follow-up period (P < 0.001). The HIV-1/HIV 2 infectivity ratio was inferred to be approximately constant over time, with estimated common value 3.55. The finding of greater HIV-1 infectivity persisted in sensitivity analyses and in covariate-adjusted analyses, with adjusted infectivity ratio estimates ranging between 3.40 and 3.86. Understanding the mechanisms by which HIV-1 infects more efficiently than HIV-2 may be useful in the development of HIV-1 vaccines. Additionally, the methodology developed here may be useful for analysing other data sets. PMID- 12590416 TI - Generalized covariance-adjusted canonical correlation analysis with application to psychiatry. AB - The lack of control over covariates in practice motivates the need for their adjustment when measuring the degree of association between two sets of variables, for which canonical correlation is traditionally used. In most studies however, there is also a lack of control over the attributes of responses for the sets of variables of interest. In particular, a portion of the response variable may be continuous and the other discrete. For such settings, the traditional partial canonical correlation approach is restrictive, since a covariate adjustment for a set of continuous variables is assumed. By ignoring the assumption of continuous variates and proceeding with a partial canonical correlation analysis in the presence of continuous and discrete variates, results in canonical correlation estimates that are not consistent. In this paper we generalize the traditional partial canonical correlation approach to covariate adjustment by allowing the response variables to contain continuous, as well as discrete, variates. The methodology is illustrated with a psychiatric application for examining which sleep variables relate to which depressive symptoms, as measured by commonly used constructs that presents with both continuous and discrete outcomes. PMID- 12590417 TI - Confidence intervals for the binomial parameter: some new considerations. AB - Several methods have been proposed to construct confidence intervals for the binomial parameter. Some recent papers introduced the "mean coverage" criterion to evaluate the performance of confidence intervals and suggested that exact methods, because of their conservatism, are less useful than asymptotic ones. In these studies, however, exact intervals were always represented by the Clopper Pearson interval (C-P). Now we focus on Sterne's interval, which is also exact and known to be better than the C-P in the two-sided case. Introducing a computer intensive level-adjustment procedure which allows constructing intervals that are exact in terms of mean coverage, we demonstrate that Sterne's interval performs better than the best asymptotic intervals, even in the mean coverage context. Level adjustment improves the C-P as well, which, with an appropriate level adjustment, becomes equivalent to the mid-P interval. Finally we show that the asymptotic behaviour of the mid-P method is far poorer than is generally expected. PMID- 12590418 TI - Generalized monotonic regression using random change points. AB - We introduce a procedure for generalized monotonic curve fitting that is based on a Bayesian analysis of the isotonic regression model. Conventional isotonic regression fits monotonically increasing step functions to data. In our approach we treat the number and location of the steps as random. For each step level we adopt the conjugate prior to the sampling distribution of the data as if the curve was unconstrained. We then propose to use Markov chain Monte Carlo simulation to draw samples from the unconstrained model space and retain only those samples for which the monotonic constraint holds. The proportion of the samples collected for which the constraint holds can be used to provide a value for the weight of evidence in terms of Bayes factors for monotonicity given the data. Using the samples, probability statements can be made about other quantities of interest such as the number of change points in the data and posterior distributions on the location of the change points can be provided. The method is illustrated throughout by a reanalysis of the leukaemia data studied by Schell and Singh. PMID- 12590419 TI - Stability of multivariable fractional polynomial models with selection of variables and transformations: a bootstrap investigation. AB - Sauerbrei and Royston have recently described an algorithm, based on fractional polynomials, for the simultaneous selection of variables and of suitable transformations for continuous predictors in a multivariable regression setting. They illustrated the approach by analyses of two breast cancer data sets. Here we extend their work by considering how to assess possible instability in such multivariable fractional polynomial models. We first apply the algorithm repeatedly in many bootstrap replicates. We then use log-linear models to investigate dependencies among the inclusion fractions for each predictor and among the simplified classes of fractional polynomial function chosen in the bootstrap samples. To further evaluate the results, we define measures of instability based on a decomposition of the variability of the bootstrap-selected functions in relation to a reference function from the original model. For each data set we are able to identify large, reasonably stable subsets of the bootstrap replications in which the functional forms of the predictors appear fairly stable. Despite the considerable flexibility of the family of fractional polynomials and the consequent risk of overfitting when several variables are considered, we conclude that the multivariable selection algorithm can find stable models. PMID- 12590420 TI - Understanding neural networks using regression trees: an application to multiple myeloma survival data by D. Faraggi, M. LeBlanc and J. Crowley, Statistics in Medicine 2001; 20:2965-2976. PMID- 12590422 TI - Political significance of knowledge in Southeast Europe. AB - The processes of globalization and transition are inevitable, full of dangers and threats, but offer enormous opportunities. Surveys of public opinion show that citizens are not aware of the fact that their countries are governed by the will of the people and a large majority considers that their country and the world are not going in the right direction. Presently, knowledge is becoming a dominant political power. This article outlines a strategy for building a knowledge-based society to minimize dangers, avoid threats, and take advantage of most of the opportunities, bringing a concrete action plan for Croatia, applicable to countries with similar history and socioeconomic structure. PMID- 12590423 TI - Nobel Prize winners for literature as palliative for scientific English. AB - Plagiarism causes a serious concern in scientific literature. I distinguish two types of plagiarism. What is routinely highlighted and discussed is the reprehensible type of stealing another author's ideas and words. This type I categorize as "heterotrophic" plagiarism. A more prevalent and less-discussed type of plagiarism is the verbatim use of same sentences repetitively by authors in their publications. This I categorize as "autotrophic" plagiarism. Though harmless per se, autotrophic plagiarism is equally taxing on the readers. The occurrence of autotrophic plagiarism is mainly caused by the lack of proficiency in the current lingua franca of science, ie, English. The writings of 22 Nobel literature laureates who wrote in English, especially their travelogues, essays, and letters to the press can be used for benefit of improving one's own vocabulary and writing skills and style. I suggest the writings of three literati -Bernard Shaw, Bertrand Russell, and Ernest Hemingway--as palliatives for autotrophic plagiarism in scientific publishing. PMID- 12590424 TI - Keeping our feet on the ground: we need to be aware of the dangers of losing focus on patients. AB - Doctors in training are susceptible to thinking themselves more exceptional than reality demonstrates. This tendency, combined with the many pressures of medical training, can lead to a break down in communication with patients. Reminding ourselves of the reality of our role could help us reflect on how we can best benefit them. One important way of doing this is to preserve the aspect of professionalism that health care systems often make difficult--communication. PMID- 12590425 TI - Analysis of European medical schools' teaching programs. AB - AIM: To compare the teaching programs of European medical schools with the minimum requirement medical curriculum recommended by the European University Association. Methods. Information on the curricula was gathered from the websites of 32 medical schools from 18 European countries. The data collected were the number of courses and proportion of each course in the study plan according to the class hours or credits. Each curriculum was compared with the standard medical curriculum recommended by the European University Association. Courses were clustered in 3 large groups: preclinical, clinical, and public health. RESULTS: The proportion of preclinical subjects was the highest at the Medical Schools in Brussels, Dublin, Milan, and Madrid, and the lowest in Athens, London, Lund, and Bucharest. The proportion of clinical subjects was the highest at the Medical Schools in London, Bucharest, and Genoa, and the lowest in Brussels, Dublin, and Milan. Croatian Medical Schools (Zagreb, Osijek, Split, and Rijeka) and the Medical School in Milan had the highest proportion of public health and humanistic subjects in their curricula. Neuroscience was found in less than half of the analyzed medical school curricula, and Psychology in about two-thirds. Nuclear Medicine course was found only in 7 out of 28 schools. Intensive Care, Anesthesiology, and Urology courses were found in less than half of the curricula. Two-thirds of analyzed curricula offered Primary Care course within the group of public health and humanistic courses. Epidemiology and Social Medicine courses were taught at more than half, and Anthropology at one-sixth of the medical schools. CONCLUSION: The lack of uniform curriculum in European medical schools makes mutual accreditation and mobility of students very difficult. Great deviations from the standard, ie, medical curriculum recommended by the European University Association, question the possibility of a quick medical curricula reform. PMID- 12590426 TI - Status of women in small academic medical communities: case study of the Zagreb University School of Medicine. AB - AIM: To analyze the proportion of women among student and teaching bodies of the Zagreb University School of Medicine between 1950 and 2000. METHODS: The data on medical school graduates from the Zagreb University School of Medicine between 1950 and 2000 were collected from the archive of the School. The data on the School's teaching staff between 1950 and 2000 were collected from the Archive of the Zagreb University. The data collected were the number of women among graduate students, grade average of medical school graduates, and the number of women within different faculty ranks among the teaching staff. RESULTS: The proportion of women among medical school graduates increased from 26% to 67% during the last 50 years. The grades showed significant inflation between 1970 and 1990. In 1990 and 2000, men had significantly higher grades than women. The proportion of female teaching staff increased during the same period from 28% to 34%. The proportion of assistants was significantly higher among female teaching staff in every studied year. The proportion of professors among male and female teaching staff increased at the same rate until 1990, when the proportion of male professors decreased because of the decrease in the absolute number of male professors. CONCLUSION: Women in Croatia are beginning to advance in academic medicine although the number of women among the teaching staff is far smaller than that of men, most probably because of delayed entrance of women into academic medicine. PMID- 12590427 TI - Peace test: is war sometimes a better solution? Survey of students of Zagreb and Mostar Schools of Medicine. AB - AIM: To investigate the differences among medical students from two medical schools, one in Zagreb, Croatia, and the other in Mostar, Bosnia and Herzegovina, in their affinity towards peaceful vs violent way of solving conflicts. METHODS: A total of 733 students from the Zagreb and 102 medical students from the Mostar University School of Medicine filled out an anonymous questionnaire during their enrollment into the next academic year. The questionnaire consisted of 10 Likert type questions with 1-5 answer scale, which were designed to give an illustration of students' attitude towards war. The test score was calculated as the sum of all answers x 2+20. The total score ranged from minimum 40 to maximum 120 points, with a higher score indicating stronger inclination toward peaceful way of solving conflicts. RESULTS: There was no difference between the mean total scores of Zagreb and Mostar students (66-/+17 and 67-/+18, respectively; p=0.744). The mean score of female students was higher than that of male students (71-/+19 vs 63-/+16; p<0.001) for the whole sample as well as for Zagreb and Mostar samples separately (p<0.001 for both). The average score of 2.3-/+0.9 per question indicated that the students' choice was mostly undecided on war-prone activities. Younger students were more war-prone than older ones (p=0.008 for age, and p=0.024 and 0.013 for comparisons between students in earlier and later academic years). Students from cities that were affected by war but not severely damaged seemed less war-prone than students from cities that were either seriously damaged or not directly affected by war (p=0.032). CONCLUSION: Women, older students, and students from cities that were under war threat but not seriously damaged showed to be more morally engaged towards peace. PMID- 12590428 TI - Influence of health risk behavior and socio-economic status on health of Slovak adolescents. AB - AIM: To investigate the role of health risk behavior, such as smoking and alcohol consumption, in the explanation of socio-economic health differences among adolescents. The hypothesis of different exposure and the hypothesis of different vulnerability were explored. METHOD: In the study carried out in 1998, the prevalence of health complaints of smokers vs non-smokers, alcohol consumers vs abstainers, and among different socio-economic groups of 2,616 Slovak adolescents (mean age, 14.9-/+0.62 years) were investigated by means of self-reported questionnaires. The adolescents were stratified according to sex and type of secondary school. RESULTS: Socio-economic disadvantage and the presence of health risk behavior were associated with greater frequency of health complaints by adolescents. Prevalence of smokers was higher in lower socio-economic groups, but no such trend was found for the prevalence of alcohol consumers. Socio-economic status and health risk behavior interactively influenced health, when socio economic status was assessed according to the mother's characteristics. Socio economic health differences between non-smokers and abstainers were not significant, unlike the differences between the smokers and alcohol consumers. The influence of health risk behavior was weaker in higher socio-economic groups. CONCLUSION: Both hypotheses, of different exposure and different vulnerability, could explain socio-economic health differences among Slovak adolescents, with different exposure playing a more important role. PMID- 12590429 TI - Use of psychoactive substances among Zagreb University medical students: follow up study. AB - AIM: To estimate the exposure of Zagreb University medical students to psychoactive substances in 2000 and compare it with data collected in 1989. METHODS: Students were surveyed in 2000 (n=775) and 1989 (n=986) by means of a self-reporting questionnaire. The 2000 survey also included 136 non-medical students. General demographic data and data on experience with psychoactive substances were collected, analyzed, and presented as percentages with 95% CI. RESULTS: The lifetime prevalence of contact with illicit drugs (mostly marijuana) among medical students in 2000 was 35% vs 14% in 1989. Repeated use was reported by 3.9% students, and 6% confirmed the experience of simultaneous use of alcohol and drugs. The lifetime prevalence of contact with psychoactive medications (mostly benzodiazepines) was 33% in 2000 vs 15% in 1989. Medications were largely used without medical supervision, with 3.5% of the students explicitly stating non-medical reasons for consumption. The prevalence of smokers was 29% in 2000, compared with 31% in 1989, and prevalence of regular alcohol consumers (on monthly, weekly, or daily basis) was 50% vs 52% in 1989. Experience with all types of substances was more present among senior students, and less among medical students than their "non-medical" peers. Regular alcohol use was associated with the experience of illicit drugs use. In 2000, 40% of medical students and 53% of "non-medical" students voted for legalization of marijuana. CONCLUSIONS: Experience with psychoactive substances is common among Zagreb medical students. Experiences with illicit drugs and psychoactive medications have substantially increased over the past decade. PMID- 12590430 TI - Treating heroin addiction: comparison of methadone therapy, hospital therapy without methadone, and therapeutic community. AB - AIM: To analyze the success of three different treatment modalities for heroin addiction: methadone therapy, hospital therapy without methadone, and therapeutic community. METHODS: Ninety heroin addicts from the city of Split, Croatia, were systematically allocated to three groups, with 30 participants each, according to the addiction treatment modality they underwent or treatment that resulted in the longest abstinence. All participants were heroin addicts for at least 2 years before the treatment. The first group was treated by methadone, the second by hospital therapy, and the third in a therapeutic community. The criterion of the treatment success was minimum two-year heroin abstinence after therapy. RESULTS: After methadone therapy, only a single participant abstained from heroin for more than 2 years, but continued to use soft drugs and alcohol. After hospital therapy, none of the participants abstained from heroin. After therapy in a therapeutic community, 9 participants abstained from heroin; 5 of them continued to use soft drugs and/or alcohol. There was a statistically significant relation between heroin abstinence and therapeutic method (chi-square=16.4236, p<0.001; Fisher's exact test=14.246, p<0.001). CONCLUSION: Treating heroin addiction in a therapeutic community may be a better treatment modality than methadone or hospital therapy. PMID- 12590431 TI - Antiandrogens in hormonal contraception limit muscle strength gain in strength training: comparison study. AB - AIM: To determine antiandrogen effects on muscle strength gain and fat-free mass increase during exercise in women using hormonal contraception with antiandrogen content. METHODS: The study included 50 women (age range, 18-30 years; mean-/+SD, 26-/+3) using hormonal contraception for at least 12 months before the beginning of the study. They were divided into two groups: "antiandrogen" group (n=26) and "estrogen-progestogen" group (n=24) groups. The subjects participated in strength training sessions 3 times a week during 16 weeks. Initially, there were no differences between the groups in fat-free mass, muscle strength, or maximum oxygen uptake. The parameters were measured before, during, and after the training period. RESULTS: After the training period, the mean increase of fat free mass was significantly greater in the estrogen-progestogen group (p<0.001). The mean gain in the muscle strength (in Newtons) was also more evident in estrogen-progestogen group (1289.0-/+17.1 N vs 101.9-/+18.5 N; p<0.001). There were no differences in the maximum oxygen consumption (VO(2)max). CONCLUSION: Antiandrogens had a negative effect on muscle strength, minimizing the effects of strength training in women. Hormonal contraception containing antiandrogens should be avoided in young exercising women. PMID- 12590432 TI - Factors influencing medical informatics examination grade--can biorhythm, astrological sign, seasonal aspect, or bad statistics predict outcome? AB - AIM: To investigate whether and to what extent various parameters, such as individual characteristics, computer habits, situational factors, and pseudoscientific variables, influence Medical Informatics examination grade, and how inadequate statistical analysis can lead to wrong conclusions. METHODS: The study included a total of 382 second-year undergraduate students at the Rijeka University School of Medicine in the period from 1996/97 to 2000/01 academic year. After passing the Medical Informatics exam, students filled out an anonymous questionnaire about their attitude toward learning medical informatics. They were asked to grade the course organization and curriculum content, and provide their date of birth; sex; study year; high school grades; Medical Informatics examination grade, type, and term; and describe their computer habits. From these data, we determined their zodiac signs and biorhythm. Data were compared by the use of t-test, one-way ANOVA with Tukey's honest significance difference test, and randomized complete block design ANOVA. RESULTS: Out of 21 variables analyzed, only 10 correlated with the average grade. Students taking Medical Informatics examination in the 1998/99 academic year earned lower average grade than any other generation. Significantly higher Medical Informatics exam grade was earned by students who finished a grammar high school; owned and regularly used a computer, Internet, and e-mail (p< or =0.002 for all items); passed an oral exam without taking a written test (p=0.004), or did not repeat the exam (p<0.001). Better high-school students and students with better grades from high-school informatics course also scored significantly better (p=0.032 and p<0.001, respectively). Grade in high-school mathematics, student's sex, and time of year when the examination was taken were not related to the grade, and neither were pseudoscientific parameters, such as student zodiac sign, zodiac sign quality, or biorhythm cycles, except when intentionally inadequate statistics was used for data analysis. CONCLUSION: Medical Informatics examination grades correlated with general learning capacity and computer habits of students, but showed no relation to other investigated parameters, such as examination term or pseudoscientific parameters. Inadequate statistical analysis can always confirm false conclusions. PMID- 12590433 TI - Attitudes towards science and alternative medicine of medical, economics and business, and electrical engineering students in Split, Croatia. AB - AIM: To assess attitudes of students at the Split University Schools of Medicine, Economics and Business, and Electrical Engineering towards science and alternative medicine. METHODS: There were 275 respondents surveyed for their opinion on science and different aspects of alternative medicine. The respondents were grouped into three major groups: medical students (83 or 30%), economics and business students (95 or 35%), and electrical engineering students (97 or 35%). Each group was composed of two subgroups: first and final year students. We used a specially designed questionnaire composed of four parts to collect students' demographic data and investigate their scientific experience and attitudes towards science and alternative medicine. ANOVA was used to test the differences among the three major groups of students and between first and final year students within each group. RESULTS: There was no significant difference among the first year students at three different schools in their scores on attitude towards science (ANOVA; F=0.001, p=0.99) and the attitude towards alternative medicine (F=2.61, p=0.08). However, there was a significant difference among the final year students at the three schools, with medical students being the most positive in their score on the attitude towards science (F=7.30, p=0.001) and least positive in their score on the attitude towards alternative medicine (F=30.7, p<0.001). There was a negative correlation between attitudes towards science and alternative medicine (r=-0.37, p<0.001) for all schools. CONCLUSION: Medical students showed the most positive attitude towards science and least positive attitude towards alternative medicine, compared with the students of economics and business or electrical engineering. The curriculum of the Medical School, unlike the curricula of other two schools, could have an impact on students' attitudes towards science. PMID- 12590434 TI - Sexual practices of undergraduate students in Tirana, Albania. AB - AIM: To assess the prevalence of sexual activity among undergraduate students at the University of Tirana, Albania, their age at first sexual intercourse, and the influence of socioeconomic factors on their sexual practices, including condom use. METHODS: An anonymous questionnaire survey was carried out among 720 undergraduate students (77% women) at the University of Tirana in October and November 2002. The questionnaire inquired about their current or previous sexual activity, age at first sexual intercourse, and use of condoms. Socio-demographic data were also collected. Multiple regression analyses were used to assess the associations between socio-demographic factors and the investigated characteristics of sexual practices of Albanian undergraduates. RESULTS: The prevalence of current and/or previous sexual activity was higher among men than women (65% vs 34%, p<0.001). Also, men engaged in sexual activity at earlier age than women (mean age, 17.9 years; 95% confidence interval [CI] 17.6-18.2 for men vs 18.8 years; 95% CI, 18.6-19.1 for women). After adjustment for covariates, both parental high education and high income level were strongly and significantly associated with students' engagement in sexual activity (odds ratio [OR], 3.46; 95% CI, 1.11-9.80 for education; and OR, 2.11; 95% CI, 1.14-3.92 for income) and the likelihood of consistent condom use by students who reported having current or past sexual relations (OR, 8.4; 95% CI, 3.9-18.3 for education; and OR, 7.0; 95% CI, 1.9-26.4 for income). CONCLUSION: Parental education and income level are strongly associated with sexual behavior (engagement in sexual activity and consistent condom use) of Albanian undergraduates in Tirana. PMID- 12590435 TI - Knowledge and attitude of undergraduate students towards sexually transmitted infections in Tirana, Albania. AB - AIM: To assess the knowledge and attitude of undergraduate students in Tirana, Albania, towards sexually transmitted infections (STI). METHODS: A sample of 729 students (76% women) at the University of Tirana were surveyed by the use of an anonymous questionnaire with 10 multiple-choice questions testing their knowledge and 27 statements testing their attitude towards STI (adapted to a 1-5 Likert scale, with a low score indicating poor attitude). Socio-demographic data were also collected. The survey was carried out in October and November 2002. Multiple regression analysis was used to assess the associations between socio-demographic factors on the one hand and attitude towards and knowledge about sexually transmitted infections, on the other. RESULTS: Men had poorer knowledge and attitude toward STI than women (beta= -4.59, p<0.001 for attitude, and beta= 0.38, p=0.006 for knowledge). Higher parental education and urban origin were strongly associated with better STI knowledge. After adjustment for age, sex, marital status, religion, income, and number of siblings, students whose parents had low and middle education level had a mean STI knowledge score of <1.16 (95% confidence interval [CI], 0.85-1.47) and <0.98 (95% CI, 0.74-1.23), respectively, which was significantly lower than the score of students with highly educated parents (p<0.001 for linear trend). Also, students born in rural areas had significantly lower mean STI knowledge score (<0.70, p=0.003) than students born in urban areas. CONCLUSION: Parental education and origin are strongly associated with knowledge and attitude of undergraduate students in Tirana towards sexually transmitted infections. PMID- 12590436 TI - Is there grade inflation at medical schools? Case study of the Zagreb University School of Medicine. AB - AIM: To investigate if students' grades at the Zagreb University School of Medicine increased since the establishment of the School in 1917. METHODS: In this retrospective descriptive study, we analyzed student sex, the length of studying, average of all grades, and grades from 5 major courses--anatomy, physiology, pathology, internal medicine, and surgery. The passing grades at the Zagreb University range from 2 (sufficient) to 5 (excellent). We analyzed data for 2,861 students from 9 representative classes, enrolled in 1920, 1930, 1940, 1950, 1960, 1970, 1980, 1985, and 1990. RESULTS: The number of female students constantly increased up to 1970 and hereafter the female to male ratio remained stable, 60:40. The percentage of enrolled students who graduated from the School increased from 1920 to 1940 and from 1960 to 1985. Between 1940 and 1960, the percentage of students who graduated was lower than 50%. There was a continuous increase in grades during the investigated period (p<0.001), except for students enrolled in 1960, who had lower grades than those enrolled in 1950. Students who enrolled in 1990 also had lower grades than those enrolling in 1985. Grades from the individual courses mostly followed the increasing trend of total grades. CONCLUSION: There has been an increasing trend in grades at the Zagreb University School of Medicine since its establishment in 1917. PMID- 12590437 TI - Learning through the Community Service. AB - AIM: To present a model of teaching general practice to medical students as a part of care for refugees in Mostar, Bosnia and Herzegovina. METHOD: With an international support, 33 medical students (from the third study year on) participated in a total of 51 field visits to 4 refugee camps near Mostar, Bosnia and Herzegovina, over a period of two years. Some students made more than 30 visits. Together with residents in family medicine at the Mostar University Hospital, the students performed physical examinations and small interventions, and distributed packs of different pharmaceutical agents to refugees. At the end of the project, participants were surveyed to assess the benefits of the program. RESULTS: Thirty out of 33 participating students responded to the survey. Fourteen students said that "the opportunity to do something, however small, for the people" was the main benefit from the project, whereas 5 thought that meeting "a real patient" early in their medical studies was very beneficial. The students assisted in 1,302 physical examinations, an average of 25-/+8 examinations per visit. The most requested physical examination was blood pressure measurement (52%), followed by chest auscultation (24%) and psychological counseling (16%). They also helped with a total of 320 minor procedures, an average of 6-/+2 per visit. Among the procedures, wound care and dressing were most common (63.4%), followed by nursing care procedures (20%) and minor surgical procedures (suture removal, abscess drainage, and bed sores debridement) in 6.9% of the cases. The students helped distribute 1,917 packs of pharmaceuticals: analgesics (52.5%), antihypertensive drugs (16.1%), antidiabetics (11.2%), cardiovascular drugs (10.9%), vitamins (5.6%), and dermatological remedies (3.6%). CONCLUSION: During the first two years of the project, we achieved three objectives. Residents in family medicine had a chance to practice and gain experience as group leaders, health promoters, and teachers. Medical students were exposed to a demanding outpatient environment early in their medical training. Refugees in three camps received health care and attention they needed so much. PMID- 12590438 TI - Risks and benefits of genetically modified maize donations to southern Africa: views from Malawi. AB - In 2001 and 2002, many countries in the Southern African Development Community (SADC) have suffered from severe food shortages resulting in an estimated 14 million people facing starvation due to inadequate quantities of the staple maize. The international community's response has been the donation of foodstuffs, including genetically modified maize. Reactions of the recipient countries of Zambia, Zimbabwe, and Malawi have been different. Zambia appealed to the donors not to send genetically modified maize, whereas Malawi accepted the maize donations. Malawi is currently facing many public health challenges because 10% of its 10-million population is HIV-positive, maternal mortality rate has almost doubled between 1992 and 2000, and there are also an estimated 1 million orphans due to HIV/AIDS. In the European Union, genetically modified maize falls under "Novel Foods" and its marketing and distribution are strictly regulated by law. This has never been the case in the southern African countries. In this article, we discuss the ethical challenges associated with genetically modified maize donations to southern Africa. Although genetically modified food offers a way to avoid many adverse effects of food shortages, we believe that some of the ethical questions of genetically modified food donations should be solved first, under the leadership of the donor countries and partnership of the developing countries. There are fears that consummation of genetically modified maize could have adverse health effects. These fears must be addressed if the confidence of developing countries in the donor community is to be maintained. PMID- 12590439 TI - Publishing scientific papers based on Master's and Ph.D. theses from a small scientific community: case study of Croatian medical schools. AB - AIM: To evaluate publishing activity of medical doctors after they have obtained Master's or Ph.D. degree at the Rijeka and Zagreb University Schools of Medicine in Croatia, and establish the number of journal articles based on these theses. METHODS: Data on Master's and Ph.D. theses defended at the Rijeka and Zagreb University Schools of Medicine in the 1990-1999 period were collected by hand search of the archive. MEDLINE and Current Contents databases were searched for journal articles resulting from the theses. RESULTS: During the 10-year period, 1,535 Master's and 634 Ph.D. theses were defended at the Rijeka and Zagreb University Schools of Medicine (253 Master's and 138 Ph.D. theses from Rijeka and 1,282 Master's and 496 Ph.D. theses from Zagreb). There were 201 (14%) Master's and 218 (34%) Ph.D. theses that resulted in articles published in journals indexed in MEDLINE (13% of Master's and 11% of Ph.D. theses from Rijeka, and 14% of Master's and 41% of Ph.D. theses from Zagreb). Also, 97 (6%) Master's and 129 (20%) Ph.D. theses that resulted in articles published in Current Contents journals (8% of Master's and 6% of Ph.D. theses from Rijeka, and 6% of Master's and 24% of Ph.D. theses from Zagreb). There was no significant difference between the two Universities with respect to published articles based on Master's theses, but there were significantly more articles from Ph.D. theses in Zagreb (p<0.001). Most of the theses resulted in a single publication (95%), 19 (5%) in 2, and 2 in 3 publications. Out of all 453 journal articles, 31% were published in Croatian and 69% in international journals. CONCLUSION: Most Croatian Master's and Ph.D. theses are not made available to the scientific community. There should be more institutional effort directed at the stimulation of postgraduate students to publish their scientific work. PMID- 12590440 TI - The secret. PMID- 12590441 TI - Introducing "quality of care" to medical students. PMID- 12590442 TI - Fluorescence polarization immunoassay based on a monoclonal antibody for the detection of the organophosphorus pesticide parathion-methyl. AB - A fluorescence polarization immunoassay (FPIA) based on a monoclonal antibody for the detection of parathion-methyl (PM) was developed and optimized. Fluorescein labeled PM derivatives (tracers) with different structures were synthesized and purified by thin-layer chromatography. The influence of immunogen and tracer structures on the assay characteristics was investigated. PM concentration determinable by the FPIA ranged from 25 to 10000 ppb. The detection limit was 15 ppb. Methanol extracts of vegetable, fruit, and soil samples were diluted 1/10 for the analysis. Recovery in spiked samples averaged between 85 and 110%. The method developed is characterized by high specificity and reproducibility (CV ranged from 1.5 to 9.1% for interassay and from 1.8 to 14.1% for intra-assay). The FPIA method can be applied to the screening of food and environmental samples for PM residues without complicated cleanup. PMID- 12590443 TI - Identification on commercialized products of AFLP markers able to discriminate slow- from fast-growing chicken strains. AB - The European chicken meat market is characterized by numerous quality marks: "Label de Qualite Wallon" in Belgium, "Label Rouge" in France, denominations of geographical origin, organic agriculture, etc. Most of those certified productions have specifications requiring the use of slow-growing chicken strains. The amplified fragment length polymorphism (AFLP) technique has been used to search molecular markers able to discriminate slow-growing chicken strains from fast-growing ones and to authenticate certified products. Two pairs of restriction enzymes (EcoRI/MseI and EcoRI/TaqI) and 121 selective primer combinations were tested on individual DNA samples from chicken products essentially in carcass form that were ascribed as belonging to either slow- or fast-growing strains. Within the resulting fingerprints, two fragments were identified as type-strains specific markers. One primer combination gives a band (333 bp) that is specific for slow-growing chickens, and another primer pair generates a band (372 bp) that was found to be characteristic of fast-growing chickens. The two markers were isolated, cloned, and sequenced. The effectiveness and the specificity of the two interesting determinants were assessed on individuals of two well-known strains (ISA 657 and Cobb 500) and on commercialized products coming from various origins. PMID- 12590444 TI - Simultaneous HPLC analysis of alpha-tocopherol and cholesterol in fresh pig meat. AB - A method has been developed for simultaneously determining alpha-tocopherol and cholesterol in fresh pig meat by HPLC. It allows a reduction in the number of analyses and brings savings in time and materials. The unsaponifiable fraction is extracted following the modified method of Liu et al. (Liu, Q.; Scheller, K. K.; Schaefer, D. M. Technical note: A simplified procedure for vitamin E determination in beef muscle. J. Anim. Sci. 1996, 74, 2406-2410). The modifications introduced are the use of nitrogen atmosphere during the extraction, the addition of an antioxidant in the organic extraction phase, and the use of alpha-tocopherol itself as an internal standard. There is then a chromatographic analysis which allows the separation of the two compounds in question. To identify and quantify, two different detectors are used in series: the first is a fluorescence detector (alpha-tocopherol), and the second is a light-scattering detector (cholesterol). The technique shows sufficient sensitivity to determine the normal levels of alpha-tocopherol and cholesterol in meat, with recovery percentages of 78% and 97%, respectively. The average amount of alpha-tocopherol and cholesterol in samples from pig Longissimus dorsi muscle analyzed using this method is 1.8 and 620 mg/kg of fresh meat, respectively. PMID- 12590446 TI - Determination of 4,4'-dinitrocarbanilide (DNC), a component of Nicarbazin, in Canada goose (Branta canadensis) eggshells using high-performance liquid chromatography. AB - A method was developed using high-performance liquid chromatography to assay 4,4' dinitrocarbanilide (DNC), the active ingredient in Nicarbazin, in eggshells collected from Canada geese fed a formulated feed fortified with Nicarbazin at doses of 0, 125, 250, and 500 microg/g. The method was developed using chicken eggshells fortified with DNC. The method was used to quantify DNC in both the shell-associated membranes and the calcified shell extracellular matrix. These values were compared to those obtained for a composite sample consisting of both the membranes and the calcified shell extracellular matrix. The validated method was used to quantify DNC in eggshells from geese fed fortified feed to ascertain the effect of Nicarbazin feed concentration on shell DNC concentration. DNC levels in the eggshells were highly correlated with feed dose. PMID- 12590445 TI - Simultaneous determination of Cd(II), Cu(II), Pb(II), and Zn(II) in citrus essential oils by derivative potentiometric stripping analysis. AB - Citrus essential oils are widely used in the food, cosmetics, and pharmaceutical industries, so the determination of heavy metals content is of great importance to guarantee their quality. The present work deals with the quantification of Cd(II), Cu(II), Pb(II), and Zn(II) in different varieties of citrus essential oils, using derivative potentiometric stripping analysis. Two different metals extraction procedures, involving concentrated hydrochloric acid treatment and acid-alcoholic dissolution, are tested on lemon, mandarin, sweet orange, and bergamot essential oils, and they give very similar results. Cd(II), Cu(II), Pb(II), and Zn(II) recovery tests spanned from 95 to 100.50%, providing evidence that metals quantification remained unaffected by the cleanup steps of the two procedures. The repeatability of the hydrochloric acid extraction method, applied on different varieties of essential oils, is >95.00% for Cd(II), Cu(II), Pb(II), and Zn(II), whereas the repeatability of the acid-alcoholic dissolution method is >93.00% for Cu and Cd only in lemon oil. Detection limits obtained for the four analytes, using both procedures, ranged from 0.10 to 0.98 ng g(-)(1) in lemon, mandarin, sweet orange, and bergamot essential oils. PMID- 12590447 TI - Optimization of solid phase microextraction analysis for the headspace volatile compounds of parmesan cheese. AB - Optimum conditions of solid phase microextraction (SPME) analysis of the headspace volatile compounds of Parmesan cheese in airtightly sealed 100-mL bottles were developed. The coefficient of variation of SPME analysis on the headspace volatile compounds of Parmesan cheese was 2%. The reproducibility of SPME was improved by a combination of sampling at -10 degrees C, controlling the sample temperature, and uniform magnetic stirring of samples during equilibrium and isolation steps. The sensitivity of SPME increased by 125% in total peak areas by a combination of 40 min of sonication and 25% (w/v) sodium phosphate solution, compared with that of samples containing deionized water only (P < 0.05). The addition of salt solution or sonication treatment in samples increased the headspace volatile compounds of cheese quantitatively without producing any new volatile compounds. PMID- 12590448 TI - Fast determination of catechins and xanthines in tea beverages by micellar electrokinetic chromatography. AB - Antioxidant properties and stimulating effects of green tea are related to its content of cathechins and xanthines; tea quality evaluation is based on organoleptic tests and on the presence of those components. In this work, by a MEKC method, eight cathechins and three xanthines were quantified in some tea based beverages. The best separation was realized using a phosphate-borate running buffer, with sodium dodecyl sulfate as micellar agent. A 40 cm capillary, a temperature of 29 degrees C, a voltage of 30 kV, and UV detection at 200 nm were used. The method showed a very good sensitivity (limit of detection ranging from 0.0011 to 0.0051 microg/mL) and was applied to real tea samples to characterize their antioxidant content. Statistical studies were performed and showed a satisfactory reliability of the data. PMID- 12590449 TI - Multiresidue pesticide analysis in wines by solid-phase extraction and capillary gas chromatography-mass spectrometric detection with selective ion monitoring. AB - A method was developed to determine pesticides in wines. The pesticides were extracted from the wine using solid-phase extraction on a polymeric cartridge, and the coextractives were removed with an aminopropyl-MgSO(4) cartridge. Analysis was performed using capillary gas chromatography with electron impact mass spectrometric detection in selective ion monitoring mode (GC-MSD/SIM). Three injections are required to analyze all 153 organohalogen, organonitrogen, organophosphate, and organosulfur pesticides and residues. Pesticides were confirmed by retention times of the target ions and three qualifier-to-target ion ratios. Detection limits for most of the pesticides were less than 0.005 mg/L, and quantitation was determined from approximately 0.01 to 5 mg/L. Spike recoveries were performed by fortifying red and white wines at 0.01 and 0.10 mg/L. At the 0.01 ppm level, the spike recoveries were greater than 70% for 116 and 124 pesticides (out of 153) in red and white wines, respectively, whereas at the higher spike concentration of 0.10 mg/L, the recoveries were greater than 70% for 123 and 128 pesticides in red and white wines, respectively. The recoveries of less than 70% were most likely from pesticide polarity or lability, resulting in the inefficient adsorption of the pesticide to the polymeric sorbent, ineffective elution of the pesticide from the sorbent, or thermal degradation of the pesticide under GC-MSD conditions. PMID- 12590450 TI - Analysis of strigolactones, germination stimulants for striga and orobanche, by high-performance liquid chromatography/tandem mass spectrometry. AB - A simple and rapid analytical method for strigolactones, germination stimulants for the root parasitic weeds witchweed (Striga spp.) and broomrape (Orobanche spp.), has been developed using high-performance liquid chromatography connected to tandem mass spectrometry (LC/MS/MS). The natural strigolactones (strigol, sorgolactone, orobanchol, and alectrol) were clearly separated and identified by LC/MS/MS. As low as 0.1 pg/microL of strigol and 0.5 pg/microL of sorgolactone could be quantified, whereas 1 pg/microL was needed for the quantification of orobanchol (S/N > 10). Using this method, it was found that red clover produces orobanchol and alectrol but not strigol. The roots of red clover seedlings were found to produce 13, 70, 58, and 65 pg of orobanchol/plant 1, 2, 3, and 4 weeks after germination, respectively. PMID- 12590451 TI - Identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets by polyclonal antibody-based enzyme linked immunosorbent assay. AB - An indirect enzyme-linked immunosorbent assay (ELISA) has been developed for the species identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets. The assay was performed in two different formats, microtiter plates and immunostick tubes, and uses polyclonal antibodies raised in rabbits against muscle-soluble proteins of grouper (anti-GSP), wreck fish (anti-WSP), and Nile perch (anti-PSP). The antibodies were made species-specific by blocking them with the heterologous soluble muscle proteins. Immunorecognition of polyclonal antibodies adsorbed to their specific fish samples was made with swine antirabbit immunoglobulins conjugated to the enzyme horseradish peroxidase. Subsequent enzymatic conversion of the substrate allowed unequivocal identification of the species studied. PMID- 12590452 TI - Characterization of the anthocyanin fraction of sicilian blood orange juice by micro-HPLC-ESI/MS. AB - Anthocyanins of red orange juices were analyzed by micro-HPLC coupled on-line with an MS detector equipped with an ESI source. The use of microcolumn HPLC greatly enhanced detection performance, allowing direct identification of the anthocyanins present in the orange juices. The use of a soft ionization technique allows detection of the molecular ions of the aglycons. Eight components were identified, five of them for the first time in red orange juice. Three additional anthocyanins were detected, of which only the aglycon was identified. PMID- 12590453 TI - Isolation of phytotoxic compounds from tree-of-heaven (Ailanthus altissima swingle). AB - The aqueous root extract of Ailanthus altissima showed allelopathic activity against radish (Raphanus sativus L. cv. "Saxa"), garden cress (Lepidium sativum L.), and purslane (Portulaca oleracea L.) seeds. A bioassay-oriented purification of active extracts, chromatographic fractions, and compounds demonstrated dose dependent activity on germination and radicle growth of test seeds; radish seed was the most sensitive to allelochemicals. Active compounds have been isolated: ailanthone, ailanthinone, chaparrine, and ailanthinol B (quassinoid derivatives); the alkaloid 1-methoxycanthin-6-one is not active. The compound with greatest inhibitory activity is ailanthone. The data obtained suggest a possible use of tree-of-heaven root extracts or of its active constituents as natural herbicides. PMID- 12590454 TI - Identification and stereochemical characterization of lignans in flaxseed and pumpkin seeds. AB - Phytoestrogens of the lignan type are widely distributed in plant-derived food items and are believed to protect against hormone-dependent cancer. The richest known dietary source of lignans is flaxseed. Flaxseed has been reported to contain glycosides of secoisolariciresinol as the major lignan, together with small amounts of matairesinol, isolariciresinol, and pinoresinol. Secoisolariciresinol, but none of the other lignans, has so far been identified in pumpkin seeds. In the present study, two different methods for the hydrolysis of lignan glycosides are compared. Artifact formation and loss of lignans under acidic conditions were observed. Lariciresinol was identified by GC-MS analysis in two different types of flaxseed (Linum usitatissimum L. and Linum flavum L.) and in pumpkin seeds (Cucurbita pepo L.) for the first time. Likewise, the novel lignan demethoxy-secoisolariciresinol was tentatively identified in the flaxseed samples. Stereochemical analysis by chiral HPLC of several lignans isolated from flaxseed showed that secoisolariciresinol, matairesinol, and lariciresinol consisted predominantly of one enantiomer. PMID- 12590455 TI - Influence of ethylene inhibition by 1-methylcyclopropene on apricot quality, volatile production, and glycosidase activity of low- and high-aroma varieties of apricots. AB - Apricots of two varieties, Ceccona with strong aroma and San Castrese with low aroma but good firmness, were treated with 1 microL L(-)(1) 1-methylcyclopropene (1-MCP) for 12 h at 20 degrees C and then kept for shelf life at 20 degrees C and 85% relative humidity. 1-MCP treatment strongly inhibited ethylene production in apricots of both varieties, and softening was delayed. Fruit softening started before the rise of ethylene in air-treated apricots, which softened even when the rise of ethylene production was inhibited by 1-MCP. The softening reduction was more significant in Ceccona apricots than in San Castrese. Pectinmethylesterase (PME) activity declined in Ceccona fruit regardless of the treatment; in San Castrese, PME of air-treated fruit slightly increased, whereas in 1-MCP-treated apricots the activity declined. alpha-d-Galactosidase (alpha-gal) and beta-d galactosidase (beta-gal) activities in Ceccona apricot were significantly reduced by 1-MCP treatment, whereas in San Castrese apricot no difference in activities was observed between air- and 1-MCP-treated fruit. The pattern of beta-d xylosidase (xyl) activity in San Castrese apricot was similar to that of beta gal, showing a peak on day 4 without difference between treatments. alpha-d Mannosidase (alpha-man) activity of air-treated apricots of both varieties rose slightly, and 1-MCP treatment decreased the enzyme activity in both varieties. alpha-d-Glucosidase (alpha-glu) decreased in air-treated apricots in both varieties, and 1-MCP maintained higher activity in Ceccona fruit but not in San Castrese. Acidity decreased during postharvest ripening regardless of the treatment, whereas soluble solids content (SSC) increased in Ceccona apricot and slightly diminished in San Castrese ones without any effect by 1-MCP treatment. 1 MCP did not show any effect on apricot color; in contrast, it affected the volatiles profile, especially in Ceccona apricot, reducing the synthesis of lactones and promoting the rise of terpenols. PMID- 12590456 TI - Glabrene and isoliquiritigenin as tyrosinase inhibitors from licorice roots. AB - Tyrosinase is known to be a key enzyme in melanin biosynthesis, involved in determining the color of mammalian skin and hair. Various dermatological disorders, such as melasama, age spots, and sites of actinic damage, arise from the accumulation of an excessive level of epidermal pigmentation. The inadequacy of current therapies to treat these conditions as well as high cytotoxicity and mutagenicity, poor skin penetration, and low stability of formulations led us to seek new whitening agents to meet the medical requirements for depigmenting agents. The inhibitory effect of licorice extract on tyrosinase activity was higher than that expected from the level of glabridin in the extract. This led us to test for other components that may contribute to this strong inhibitory activity. Results indicated that glabrene and isoliquiritigenin (2',4',4 trihydroxychalcone) in the licorice extract can inhibit both mono- and diphenolase tyrosinase activities. The IC(50) values for glabrene and isoliquiritigenin were 3.5 and 8.1 microM, respectively, when tyrosine was used as substrate. The effects of glabrene and isoliquiritigenin on tyrosinase activity were dose-dependent and correlated to their ability to inhibit melanin formation in melanocytes. This is the first study indicating that glabrene and isoliquiritigenin exert varying degrees of inhibition on tyrosinase-dependent melanin biosynthesis, suggesting that isoflavenes and chalcones may serve as candidates for skin-lightening agents. PMID- 12590457 TI - Postharvest UV-C-irradiated grapes as a potential source for producing stilbene enriched red wines. AB - The use of postharvest controlled irradiation by UV pulses is proposed as a potential method to produce stilbene-enriched red wine. "Monastrell" grapes were UV-C-irradiated to increase stilbene content. The main inducible stilbenes were resveratrol and piceatannol, which are molecules with reported health-beneficial activities. The evolution of both compounds was followed in the different steps of an "analytical" traditional maceration wine-making process. The final wine made from UV-C-irradiated grapes was enriched about 2- and 1.5-fold in resveratrol and piceatannol, respectively, when compared to the control wine. In addition, no difference was detected regarding the standard enological parameters (color, acidity, etc.). It is strongly suggested that, with the use of more susceptible wine grapes to induce bioactive stilbenes upon UV-C irradiation, the stilbene-enrichment of wine can be much higher. PMID- 12590458 TI - Homology similarity analysis of sequences of lactoferricin and its derivatives. AB - A new method, homology similarity analysis (HSA), was developed to investigate homology pattern similarities of selected segments within sequences of peptides. This new approach facilitated elucidation of the structure-function relationships of lactoferricin derivatives. Helix propensity of positions 4-9 in the lactoferricin sequence was the most important in determining the antimicrobial activity of lactoferricin against Escherichia coli, followed by cationic charge pattern at positions 4-9 and 1-3. The pattern similarity of segments within sequences could be a useful tool for representing the distribution attributes of amino acid residue properties to the structure-function relationships of proteins and peptides, especially when used in conjunction with principal component similarity analysis followed by the regression version of artificial neural networks. PMID- 12590459 TI - Monitoring chemical changes of dry-cured Parma ham during processing by surface autofluorescence spectroscopy. AB - Parma hams at various processing stages were investigated by surface autofluorescence spectroscopy. Fluorescence "landscapes" of raw meat and salted (3 months), matured (11 and 12 months), and aged (15 and 18 months) Parma hams were obtained, and a three-dimensional data array (sample x emission x excitation) was used to develop a PARAFAC model including five components, which all exhibited characteristics of pure fluorophores regarding both excitation and emission spectra. The relative amount of each component related strongly to the processing stage, and sample age showed good correlation to fluorescence data (R = 0.98), with a relative error of prediction of approximately 1 month. Fluorescence measurements from samples of either semimembranosus or biceps femoris were used to predict chemical or sensory reference data, yielding good correlation for biceps femoris data, thereby enabling moisture content, sensory and instrumental color, and proteolysis value to be fairly well predicted. Overall, surface autofluorescence of Parma hams proved to hold relevant information, relating to major chemical/physical changes during processing. It is concluded that fluorescence spectroscopy has potential as an innovative method of quality control in dry-cured ham. PMID- 12590460 TI - Warm water treatment in combination with modified atmosphere packaging reduces undesirable effects of irradiation on the quality of fresh-cut iceberg lettuce. AB - Fresh-cut iceberg lettuce dipped in either 5 or 47 degrees C water for 2 min was packaged in modified atmosphere film bags and then exposed to 0, 0.5, 1, or 2 kGy gamma-radiation. Dipping cut lettuce in 47 degrees C water for 2 min prior to irradiation reduced antioxidant and phenolic accumulations induced by irradiation. Irradiation at 2 kGy increased cellular leakage and sogginess of cut lettuce dipped in both temperatures. Samples irradiated at 0.5 and 1 kGy had similar firmness and vitamin C and antioxidant contents as the controls after 14 and 21 days of storage except 1 kGy samples dipped at 47 degrees C had lower antioxidant contents than controls at 14 days of storage. Lettuce dipped at 47 degrees C and irradiated at 0.5 and 1 kGy had better overall visual quality and less tissue browning than corresponding irradiated samples dipped at 5 degrees C. These results suggest lettuce treated with warm water and irradiated at 0.5 or 1 kGy had the best sensory quality without significant loss in texture, vitamin C, or total antioxidants. PMID- 12590461 TI - Comparison of the total phenolic and ascorbic acid content of freeze-dried and air-dried marionberry, strawberry, and corn grown using conventional, organic, and sustainable agricultural practices. AB - Secondary phenolic metabolites play an important role in plant defense mechanisms, and increasing evidence indicates that many are important in human health. To date, few studies have investigated the impact of various agricultural practices on levels of secondary plant metabolites. To address this issue, the total phenolic (TP) content of marionberries, strawberries, and corn grown by sustainable, organic, or conventional cultural practices were measured. Additionally, the effects of three common postharvest processing treatments (freezing, freeze-drying, and air-drying) on the TP content of these agricultural products were also investigated. Statistically higher levels of TPs were consistently found in organically and sustainably grown foods as compared to those produced by conventional agricultural practices. In all samples, freeze drying preserved higher levels of TPs in comparison with air-drying. PMID- 12590462 TI - Solid-State 13C CP/MAS NMR studies on aging of starch in white bread. AB - The effects of storage methods and glycerol on the aging of breadcrumbs were studied using solid-state (13)C CP/MAS NMR. After baking, a shift in C(1) peaks from triplet (A-type) to singlet (V-type) was observed. Addition of glycerol reduced the carbon peak intensities of fresh and aged breads, which correlated well with the DSC amylopectin "melting" enthalpy (r(2) = 0.91). Upon storage of bread with crust in hermetically sealed containers (when moisture migrated from the crumbs to the crust), the (13)C CP/MAS NMR peak intensity increased more rapidly during aging than when the bread was stored without crust. Although addition of glycerol retarded the starch retrogradation, as observed by (13)C CP/MAS NMR and DSC, it accelerated the firming rate. Therefore, bread firming in this case was controlled not only by starch retrogradation but also by other events (such as local dehydration of the matrix or gluten network stiffening). PMID- 12590463 TI - Irradiation and modified atmosphere packaging effects on residual nitrite, ascorbic acid, nitrosomyoglobin, and color in sausage. AB - The present study was undertaken to evaluate the irradiation and modified atmosphere packaging effects on emulsion-type cooked pork sausage during storage for 4 weeks. CO(2) (100%), N(2) (100%), or 25% CO(2)/75% N(2) packaged sausage were irradiated at 0, 5, and 10 kGy, and residual nitrite, residual ascorbic acid, nitrosomyoglobin (NO-Mb), color values, and their correlation were observed. Irradiation significantly reduced the residual nitrite content and caused partial reduction of NO-Mb during storage. No difference was observed in ascorbic acid content by irradiation. Irradiation decreased the Hunter color a value of sausage. CO(2) or CO(2)/N(2) packaging were more effective for reducing residual nitrite and inhibiting the loss of the red color of sausage compared to N(2) packaging. Results indicated that the proper combination of irradiation and modified atmosphere packaging could reduce the residual nitrite in sausage with minimization of color change. PMID- 12590464 TI - Effect of germination on the carbohydrate composition of the dietary fiber of peas (Pisum sativum L.). AB - The effect of different conditions of pea germination on dietary fiber (DF) composition was studied. Insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) were subjected to acid hydrolysis, and the resultant neutral sugars, uronic acids, and Klason lignin were quantified. Germinated peas exhibited significantly higher contents of total dietary fiber (TDF) than the raw sample, due to the increases of both DF fractions. Under darkness conditions, germination exhibited the highest contents of IDF and SDF. Decreasing IDF/SDF ratios showed that the carbohydrate changes did not take place to the same extent during germination, the SDF fraction being the most affected. The detailed chemical composition of fiber fractions reveals increases of cellulose in the IDF of germinated samples, whereas SDF exhibits a decrease of pectic polysaccharides and also increases of polysaccharides rich in glucose and mannose. The DF results were corroborated by a comparative examination of the cell wall carbohydrate composition. PMID- 12590465 TI - Mechanism of simmondsin decomposition during sodium hydroxide treatment. AB - Jojoba seed meal shows appetite-suppressing activity due to the presence of simmondsin. This pharmacological activity disappears with treatment of the meal with sodium hydroxide. To elucidate this mechanism of inactivation, the reaction of simmondsin in 1 N NaOH at 20 degrees C was monitored as a function of time. The end products of the reaction as well as intermediates were isolated and identified. The half-life of simmondsin was approximately 60 min with d-glucose and 2-hydroxy-3-methoxyphenylacetonitrile as reaction end products. The reaction mechanism could be elucidated by the isolation of isosimmondsin and a simmondsin lactone derivative. Those compounds were isolated and purified by a combination of column chromatography and HPLC and identified mainly by HRMS and NMR spectroscopy. PMID- 12590466 TI - Influence of drying on the flavor quality of spearmint (Mentha spicata L.). AB - Spearmint (Mentha spicata L.) was dried using three different drying methods: oven-drying at 45 degrees C, air-drying at ambient temperature, and freeze drying. The effect of the drying method on the volatile compounds and on the structural integrity and sensory characteristics of the spice was evaluated. The volatile components from fresh and dried spearmint samples were isolated by simultaneous distillation-extraction (SDE) and analyzed by gas chromatography mass spectrometry (GC-MS). A total of 28 compounds were identified, carvone, limonene, and 1,8-cineole, in that order, being the main components in all of the samples. Oven-drying at 45 degrees C and air-drying at ambient temperature were the methods that produced the best results. An increase in monoterpenes was observed in all of the dried samples, except in the freeze-dried samples that underwent freezing at -198 degrees C. Freeze-drying resulted in substantial losses in oxygenated terpenes and sesquiterpenes. The effect of each drying method on leaf structure was observed by scanning electron microscopy. From a sensory standpoint, drying the spearmint brought about a decrease in herbaceous and floral notes together with an increase in minty odor. PMID- 12590467 TI - Radiolysis of kaempferol in water/methanol mixtures. Evaluation of antioxidant activity of kaempferol and products formed. AB - Oxidative reaction between hydroxymethyl radical ((*)CH(2)OH) and kaempferol, in methanol and methanol/water mixtures, was studied by gamma-radiolysis using a (60)Co source. Radiolysis was performed with concentrations and doses ranging from 5 x 10(-)(5) M to 5 x 10(-)(3) M and from 0.5 kGy to 14 kGy, respectively. Kaempferol degradation was followed by HPLC. Results showed that (*)CH(2)OH reacts with kaempferol at the 3-OH group and produces two depsides (K1 and K2) and other products including K3. K1, K2, and K3 were identified by NMR, LC-MS, and HRMS. The kaempferol degradation pathway leading to the K1, K2, and K3 formation is proposed. It was observed that the more water concentration in the irradiation medium increases, the more K2 concentration increases. Comprehension of food preservation is not clear because many phenomena occurring during irradiation are not established. Radiolysis of kaempferol in water/methanol mixtures helps to elucidate the phenomenon and it is possible that during the treatment of nutriments by gamma-irradiation, a series of products such as depside K2 could be formed. Antioxidant properties of kaempferol radiolysis products were evaluated according to their capacity to decrease the EPR DPPH (1,1 diphenyl-2-picrylhydrazil) signal and to inhibit superoxide radicals formed by the enzyme reaction "xanthine + xanthine oxidase". PMID- 12590468 TI - Changes in triacylglycerol composition during ripening of sea buckthorn (Hippophae rhamnoides L.) seeds. AB - Changes in the quantitative composition of triacylglycerols (TAGs) in maturing sea buckthorn (Hippophae rhamnoides L.) seeds were determined by lipase hydrolysis. As a whole, the rate of synthesis of separate TAG classes increased in proportion to both their unsaturation and relative content (weight percent) in total TAGs. Up to the 80th day of maturation, the formation of triunsaturated TAGs was predominant. Subsequently, at the terminal stage of seed ripening, the absolute content (in nanomoles per seed) of a major group of these TAGs containing linolenic and linoleic acyls decreased by approximately 7%, and the increase in the total TAG content was mainly due to the synthesis of TAG molecules including stearic and palmitic acyls in the rac-1,3 positions, as well as those containing oleate in the sn-2 position. At each maturation stage, the composition of the TAGs formed was controlled both by the composition of fatty acids available for TAG synthesis and by the rate of incorporation of a particular fatty acid into the sn-2 position of the TAGs. PMID- 12590469 TI - Use of the enantiomeric composition for the assessment of the authenticity of fruit beverages. AB - Enantiomeric compositions of chiral terpenes in commercial fruit beverages were examined by solid phase microextraction-gas chromatography (SPME-GC). Optimization of the method was accomplished on the basis of some parameters involved in the extraction, such as heating temperature and extraction time, that provided the highest peak areas, 60 degrees C and 2 min being the optimal values. With the proposed method relative standard deviation (RSD) values from three replicates ranging from 2 to 12% were obtained. The enantiomeric distribution of some terpenes remained constant, whereas other terpenes (linalool, terpinen-4-ol, and alpha-terpineol) exhibited a considerable variation among samples. This can be indicative of the eventual addition of aromas to some fruit beverages. PMID- 12590470 TI - Alkylresorcinol derivatives and sesquiterpene lactones from Cichorium spinosum. AB - One new alkylresorcinol derivative, cichoriol B, and a mixture of three other ones, cichoriols A, C, and D, were isolated from the dichloromethane extract of Cichorium spinosum, a plant that is used traditionally in the Cretan diet. The methanol extract afforded one new sesquiterpene lactone, (4R)-3,4 dihydrolactucopicrin. The structures of the new compounds were determined by spectroscopic methods, mainly by the concerted application of 1D and 2D NMR techniques (HMQC, HMBC, NOESY). PMID- 12590471 TI - Determination of 5-methyltetrahydrofolic acid and folic acid in citrus juices using stable isotope dilution-mass spectrometry. AB - A stable isotope liquid chromatography-mass spectrometry (LC-MS) method was developed for the quantitative determination of 5-methyltetrahydrofolic acid (5 MTHFA) and folic acid in a variety of commercial citrus juices. Folates were extracted from juices, and the polyglutamyl side chain of 5-MTHFA was cleaved to the monoglutamate form using rat plasma conjugase. The folates were purified on a Bond-Elut column and analyzed by LC-MS with electrospray ionization. The analytes were quantified using the (13)C(5) analogues of 5-MTHFA and folic acid as internal standards. The relative standard error of the method was 3.35% based on replicate analyses (n = 4). This method was then applied to the determination of 5-MTHFA and folic acid in a variety of citrus juices obtained from local supermarkets. It was observed that although both "store" brands and "national" brands of fresh (nonfrozen) juices contained similar concentrations of 5-MTHFA, the "store" brands of fresh juices had on average >5-fold the amount of folic acid compared to the "national" brands. In addition, the "total" folate concentrations were generally below values listed on the food label. PMID- 12590472 TI - Rapid quantitative enrichment of carnosic acid from rosemary (Rosmarinus officinalis L.) by isoelectric focused adsorptive bubble chromatography. AB - For the first time, the potent but unstable antioxidative diterpene carnosic acid could be enriched from an aqueous extract of rosemary (Rosmarinus officinalis L.) by isoelectric focused adsorptive bubble chromatography. Enrichment of carnosic acid in the foam was influenced by the pH value and the flow rate of the foam forming gas. Efficiency was highest with diluted samples at pH 4. Under these conditions, the conversion of carnosic acid to carnosol was negligible. Transfer of carnosic acid to the foam from a standard solution in the presence of saponin as surfactive substance was similar to that from the aqueous rosemary extract. PMID- 12590473 TI - Color characteristics of monascus pigments derived by fermentation with various amino acids. AB - Various pigment colors were produced by Monascus fermentations with separate addition of 20 amino acids. The color characteristics and structures of the pigment derivatives were investigated. When each amino acid was added to the fermentation broth as a precursor, pigment extracts with different hue and chroma values were obtained depending on the content ratios of yellow, orange, and red colors in the fermentation broth. The yellow and orange pigments were identical regardless of amino acid addition. The red compounds varied on the basis of the type of amino acid added. LC-MS and (1)H and (13)C NMR structural analyses confirmed that the derivative pigments contained the moieties of the added amino acids. L, a, and b values of the CIELAB color system for the derivative pigments were measured. Values of hue and chroma were then calculated. The colors of the derivative pigments were in the range of orangish red to violet red. The hydrophilicities/hydrophobicities of the derivative pigments could be predicted from their log P values, which were estimated using computer programs. PMID- 12590474 TI - Analysis of dietary supplements for arsenic, cadmium, mercury, and lead using inductively coupled plasma mass spectrometry. AB - The arsenic, cadmium, mercury, and lead contents of 95 dietary supplement products were determined using microwave digestion and high-resolution inductively coupled plasma mass spectrometry. Precision and accuracy were demonstrated by element recovery from 17 dietary supplements and replicates of 8 reference materials. The concentration ranges were as follows: arsenic, <5-3770 microg/kg; cadmium, <10-368 microg/kg; mercury, <80-16800 microg/kg; and lead, <20-48600 microg/kg. An assessment of estimated exposures/intakes of the four elements is presented. PMID- 12590475 TI - Variations in the cell wall composition of maize brown midrib mutants. AB - Most studies published thus far on the four brown midrib (bm) mutants (bm1, bm2, bm3, and bm4) in maize (Zea mays L.) have focused on one or two individual mutants, and comparisons between studies have been difficult because of variation in genetic backgrounds, maturity, and source of tissue. Detailed analyses of the stalks of the four bm single mutants and a bm1-bm2 double mutant in a common genetic background (inbred A619) revealed structural and compositional changes in their isolated cell walls and lignins compared to the wild-type inbred. 2D-NMR revealed a significant presence of benzodioxane units in the bm3 isolated lignin. 1D (13)C NMR revealed increased aldehyde levels in the bm1 and bm1-bm2 mutants compared to the wild-type inbred. The bm3 and bm1-bm2 mutants contained less Klason lignin in the isolated cell walls. The bm1, bm3, and bm1-bm2 mutants contained approximately 50% less esterified p-coumaric acid with noticeably elevated levels of ferulate in the bm3 mutant. A difference among bm mutants in the solubility of p-coumaric acid-lignin complexes during cellulase enzyme treatment was also discovered, suggesting that the bm mutations might also differ in the structural organization of lignin. PMID- 12590476 TI - Distribution of lutein, zeaxanthin, and related geometrical isomers in fruit, vegetables, wheat, and pasta products. AB - Quantitative data with regard to dietary (3R,3'R,6'R)-lutein, (3R,3'R) zeaxanthin, and their (E/Z)-geometrical isomers are scarce, and in most cases, only the combined concentrations of these two carotenoids in foods are reported. Lutein and zeaxanthin accumulate in the human macula and have been implicated in the prevention of age-related macular degeneration (AMD). The qualitative and quantitative distributions of lutein, zeaxanthin, and their (E/Z)-isomers in the extracts from some of the most commonly consumed fruits, vegetables, and pasta products were determined by HPLC employing a silica-based nitrile-bonded column. Green vegetables had the highest concentration of lutein (L) and zeaxanthin (Z), and the ratios of these carotenoids (L/Z) were in the range 12-63. The yellow orange fruits and vegetables, with the exception of squash (butternut variety), had much lower levels of lutein in comparison to greens but contained a higher concentration of zeaxanthin. The ratio of lutein to zeaxanthin (L/Z) in two North American bread varieties of wheat (Pioneer, Catoctin) was 11 and 7.6, respectively, while in a green-harvested wheat (Freekeh) imported from Australia, the ratio was 2.5. Between the two pasta products examined, lasagne and egg noodles, the latter had a much higher concentration of lutein and zeaxanthin. The levels of the (E/Z)-geometrical isomers of lutein and zeaxanthin in these foods were also determined. PMID- 12590477 TI - Comparative persistence of pesticides on selected cultivars of specialty vegetables. AB - The behavior and dissipation of several pesticides on selected cultivars of specialty vegetable crops were compared to determine appropriate preharvest intervals for compliance with maximum residue limits. To reduce application variability, a tank mix of pesticides was applied for each trial. Residues of eight pesticides applied to bok choi, Chinese broccoli, and fuzzy squash were similar for the two leafy vegetables; residues were higher than on squash because of the latter's larger mass-to-surface area ratio. Whereas residues of five of the nine pesticides applied to cabbage, Chinese cabbage, and bok choi did not differ significantly from 3 to 14 days after application, residues of cypermethrin and three fungicides were significantly higher on Chinese cabbage and bok choi. The residue deposits of the pesticides were about 50% lower on Chinese cabbage and 90% lower on cabbage than on bok choi. Dissipation of residues on different cultivars of lettuce and cabbage were compared after application of nine pesticides. With the exception of captan, residues on head lettuce were lower than on the other lettuces on day 1; cultivar differences were not significant for most of the nine applied pesticides on days 3 and 8. Generally, residues of the nine individual pesticides on storage cabbage, savoy cabbage, Chinese cabbage, and tah tsai did not differ significantly from day 1 to day 7 after application. Residue concentrations were generally significantly higher on bok choi than on the other cultivars. Residue deposits generally correlated with the rate of application; residues of captan, however, were about 50% of the predicted value. Significant differences in deposit and dissipation were observed among cultivars and pesticides, with dramatic initial decreases for diazinon and parathion. Because of their higher exposed surface area-to-mass ratios, leafier crops had higher residue concentrations than head varieties. Residues on lettuce cultivars were higher than on the cabbages. The results clearly indicate that structure significantly affects residue deposit and dissipation, and pesticide recommendations cannot always be extended to specialty crops without an investigation of the changes in preharvest intervals to prevent violations of the maximum residue limits. PMID- 12590478 TI - Biotransformation of an organochlorine insecticide, endosulfan, by Anabaena species. AB - This study assesses the role of the blue-green algal species present in the soil in the dissipation of endosulfan and its metabolites in the soil environment. Two Anabaena species, Anabaena sp. PCC 7120 and Anabaena flos-aquae, were used in this study. Anabaena sp. PCC 7120 produced three principal biotransformation compounds, chiefly endosulfan diol (endodiol), and minor amounts of endosulfan hydroxyether and endosulfan lactone. Trace amounts of endosulfan sulfate were detected. In comparison, the biotransformation of endosulfan by Anabaena flos aquae yielded mainly endodiol with minor amounts of endosulfan sulfate. An unknown compound was produced up to 70% from endosulfan spiked in the medium inoculated by A. flos-aquae after 8 days of incubation. Therefore, the endosulfan fate was dependent on the species. Within 1 day of incubation, two Anabaena species produced low amounts of beta-endosulfan after application of alpha endosulfan. These results suggest the presence of isomerase in the Anabaena species. Further studies using a fermentor to control the medium pH at 7.2 to minimize chemical hydrolysis of endosulfan revealed a major production of endodiol with minor amounts of endosulfan sulfate and the unknown compound. These results showed that the production of the unknown compound might be dependent on the alkaline pH in the medium and that the production of endodiol by A. flos aquae might be biologically controlled. This study showed that two algal species could contribute in the detoxification pathways of endosulfan in the soil environment. PMID- 12590479 TI - Determination of degradation products and pathways of mancozeb and ethylenethiourea (ETU) in solutions due to ozone and chlorine dioxide treatments. AB - The objective of the present study was to determine the degradation products of mancozeb and ethylenethiourea (ETU) and elucidate the possible degradation pathways in solution as a result of chemical oxidation using ozone and chlorine dioxide. This study was developed in a solution at 100 ppm of mancozeb and ETU concentration over the course of 60 min. Two different oxidizing agents used in this study were (1) ozone at 3 ppm and (2) chlorine dioxide at 20 ppm. Ozone was continuously provided throughout the course of the reaction. Degradation products were detected with high-resolution GC-MS. The total analysis time was 4 min per sample combined with rapid GC separation and time-of-flight mass spectrometry (TOFMS). Hydrolysis of mancozeb led to m/z 144 ion fragmentation, which is 5 imidazoledithiocarboxylic acid, as a major degradation product. ETU showed M(+) 102, which corresponds to its mass, indicating this compound was stable in distilled water and did not undergo hydrolysis during 60 min. The average retention times of mancozeb and ETU were approximately 181-189 and 210-230 s, respectively. Ozonation of mancozeb produced ETU as a major product. Treatment of ETU with ozone produced several degradation compounds. From prolonged ozonation, the CS(2) or CS group was removed. Overall, several byproducts identified were M(+) 60, M(+) 84, M(+) 163, M(+) 117, and M(+) 267 by ozone and M(+) 117, M(+) 86, and M(+) 163 by chlorine dioxide treatment. Several of these have been reported, but others have never been reported previously. PMID- 12590480 TI - Fipronil metabolism and dissipation in a simplified aquatic ecosystem. AB - Several phenylpyrazole derivatives are selective inhibitors of chloride channel activities in insects. In this chemical family, fipronil is a powerful insecticide now widely used for several purposes. The dissipation of this molecule in a simplified aquatic ecosystem has been studied for 3 months, using (14)C-labeled fipronil. The main features of the complex process leading to fipronil transformation in this system were the following. The fipronil aqueous solution was submitted to two chemical transformations: the photodependent desulfuration of the side chain bound to the 4-position of the heterocyclic ring and the chemical hydrolysis of the nitrile function bound to the 3-position. Fipronil, rapidly transferred from the water solution to the organic matter, was protected from the previously mentioned chemical transformations but evolved to give two main metabolites, which were either reduced or oxidized in the side chain on the 4-position. These derivatives were powerful insecticides as shown by LC(50) measurements on Aedes aegypti larvae (LC(50) for CF(3)-S-R and CF(3)-SO(2) R = 8.8 nM). During the course of this experiment, nitrile hydrolysis took place slowly, originating either from the chemical hydrolysis in the aqueous solution or from enzymatic hydrolysis inside the microbial biomass. The fipronil-amide (3 NH(2)-CO-R') derivative, although much more polar than fipronil itself, was mostly bound to the organic matter. Other more polar derivatives were also detected but in very small amounts. No (14)CO emission was observed during the experiment. PMID- 12590481 TI - Occurrence and distribution of organochlorine pesticides (OCPs) in tomato (Lycopersicon esculentum) crops from organic production. AB - Organochlorine pesticides (OCPs) were quantified by GC-ECD in tomato (Lycopersicon esculentum) during a vegetation period. Plants were harvested at 15, 60, and 151 days after seed germination. Leaves, stem, roots, and fruit (peel and flesh) were analyzed separately. The results showed that tomato plants were able to accumulate OCPs from soils, and a trend to reach the equilibrium among tissues at mature stages was also observed. Endosulfans comprised the main OCP group, probably due to its spray during summer months in the surrounding areas. Banned pesticides such as DDTs, heptachlor, and dieldrin were found. OCPs levels in the fruit were below the maximum residues limits (MRL) considered by the Codex Alimentarius. DDE/DDT and alpha-/gamma-HCH ratios of <1 would indicate recent inputs of DDT and lindane in the environment. The occurrence of OCPs in the study farm, where agrochemicals have never been used, is a result of atmospheric deposition of those pesticides. PMID- 12590482 TI - Xenobiotic organic compounds in runoff from fields irrigated with treated wastewater. AB - Investigations of agricultural nonpoint source pollution typically focus on a relatively narrow range of targeted toxic and biostimulatory compounds (e.g., specific pesticides, nutrients). Regular application of numerous other organic compounds to agricultural fields in pesticide formulations, irrigation water, soil amendments, and fertilizers may result in their transport into surface waters via runoff. We examined whether potentially toxic dissolved and particle associated "nontarget" organic compounds were present in surface runoff from agricultural fields irrigated with disinfected tertiary recycled water or wastewater effluent-dominated streamwater. Gas chromatographic-mass spectrometric analyses of filtered runoff samples revealed the presence of numerous nontarget compounds of potential toxicological significance including pesticide transformation products, pesticide adjuvant chemicals, plasticizers, flame retardants, pharmaceuticals, and personal care product ingredients. Although the toxicity of many of these compounds is poorly characterized, some may elicit subtle but profound toxicological effects. Agricultural runoff also represented a source of allochthonous natural organic matter to the stream system. PMID- 12590483 TI - Contribution of benzenemethanethiol to smoky aroma of certain Vitis vinifera L. wines. AB - Benzenemethanethiol, a volatile thiol with a strong empyreumatic aroma reminiscent of smoke, has been identified in boxwood (Buxus sempervirens L.) as well as in red and white Vitis vinifera L. wines. The perception threshold in a model hydroalcoholic solution is approximately 0.3 ng/L. All of the wines analyzed for this study contained this compound in concentrations of several dozen nanograms per liter. The Chardonnay wines had 30-40 ng/L. Sensory discrimination between a wine containing 7 ng/L benzenemethanethiol and the same wine with an additional 4 ng/L is very significant; the difference in smell is described as "empyreumatic". This compound can therefore significantly contribute to the aroma of certain wines (Sauvignon Blanc, Semillon, Chardonnay, etc.) containing concentrations as high as 30-100 times higher than their perception threshold. PMID- 12590484 TI - Identification of key odorants related to the typical aroma of oxidation-spoiled white wines. AB - The oxidative degradation of white wines rapidly leads to a loss of their sensorial qualities. The identification of the most important descriptors related with oxidation-spoiled wine was performed by a trained sensory panel. The terms selected were "honey-like", "farm-feed", "hay", and "woody-like". By gas chromatography-olfactometry analysis three aromatic zones related to these descriptors in the oxidation-spoiled white wines could be determined. Comparison of the aroma extract dilution analysis aromagrams of oxidation-spoiled white wines and a nonspoiled wine showed the highest values of dilution factors were attributed to 3-(methylthio)propionaldehyde, phenylacetaldehyde, 1,1,6-trimethyl 1,2-dihydronaphthalene (TDN), and 4,5-dimethyl-3-hydroxy-2(5H)-furanone (sotolon). A "forced aging" experiment was implemented to simulate the typical oxidation-spoiled aroma. Samples rated with the highest score in the ranking test were also those that presented the highest concentration of these four molecules. To test the sensory impact of these substances, a normal wine (unspoiled) was spiked with these molecules (with the exception of TDN) singly and in combination, and the similarity value (SV) between samples and the oxidation spoiled white wines was then determined. The highest value from the similarity tests was 5.4 when the three compounds were added simultaneously; 3 (methylthio)propionaldehyde alone was found to be responsible for 3.6, suggesting that, among the molecules studied, it is the most important contributor to the typical aroma of an oxidation-spoiled white wine. PMID- 12590485 TI - Volatiles from leaves, fruits, and virgin oil from Olea europaea Cv. Olivastra Seggianese from Italy. AB - The volatiles produced by leaves and fruits of Olea europaea cv. Olivastra Seggianese have been analyzed in two different phenological stages. Furthermore, the volatiles of the virgin olive oil obtained from ripe fruits has been characterized. The volatiles were sampled by means of two different techniques: hydrodistillation and SPME. Differences were observed between the two different collection times, the different organs, and sampling techniques. The major constituents were often aldehydes, particularly (E)-2-hexenal (9.8-48.0%); however, also many terpenoids have been identified, mainly (E,E)-alpha-farnesene (0.2-27.0%), linalool (0-3.6%), beta-caryophyllene (0-8.1%), and valencene (0 2.5%). This is the first investigation on this cultivar. PMID- 12590486 TI - Analysis of volatile compounds of tapereba (Spondias mombin L.) and caja (Spondias mombin L.) by simultaneous distillation and extraction (SDE) and solid phase microextraction (SPME). AB - Volatile compounds of caja and tapereba fruits, both classified as Spondias mombin, but from different geographic origins, were extracted (and analyzed) using solid phase microextraction (SPME) and simultaneous distillation and extraction (SDE). Forty-eight compounds were identified in tapereba and 47 in caja by SPME using a DVB/CAR/PMDS fiber. (E)-Caryophyllene (18.7%), ethyl butyrate (10.0%), and ethyl hexanoate (7.0%) were the most abundant components in tapereba volatiles extracted by SPME, whereas myrcene (41.1%) and beta phellandrene (8.5%) were the major compounds in caja. In the tapereba SDE extract, 46 substances were identified, and (Z)-caryophyllene (13.2%) and limonene (9.5%) were predominant. From the 42 substances found in the SDE extract of caja, the major components were myrcene (38.0%) and p-cymene (6.2%). The two fruits showed similar chromatograms upon the use of SDE and SPME. These methods made it possible to determine 30 identical components in both fruits by using SDE and 32 by using SPME. PMID- 12590487 TI - Behavior of flavor compounds in model food systems: a thermodynamic study. AB - Physicochemical parameters, such as hydrophobicity, water solubility, and volatility, of four flavor compounds (ethyl acetate, ethyl butyrate, ethyl hexanoate, and 2-pentanone) were determined. The amount of flavor compounds released from different model matrices (mineral water, purified triolein, an oil in-water emulsion, a carbohydrate matrix, and a complex matrix containing lipids and carbohydrates) into the gaseous phase was determined at thermodynamic equilibrium, at 37 degrees C, by static headspace gas chromatography. The degree of interaction between the flavor compounds and the matrix components was shown by measuring the percentage retention using the water matrix as the reference. The partition of flavor compounds was principally dependent on their hydrophobicity. Physicochemical interactions that occurred in the different media led to different degrees of flavor retention. An impact of fat on flavor retention was demonstrated when a water matrix and an oil-in-water matrix or carbohydrate and complex matrices were compared. A carbohydrate impact on flavor compound retention was also detected, which was evident even in the presence of lipids. PMID- 12590488 TI - Essential oils from plants and in vitro shoots of Hypericum androsaemum L. AB - The essential oil yields obtained by hydrodistillation of the aerial parts of Hypericum androsaemum cultivated plants varied from 0.94 to 4.09 mg/g of biomass dry weight, depending of the harvest time. The respective analyses performed by gas chromatography and gas chromatography-mass spectrometry revealed more than 80 compounds, 72 of which were identified. Most of the compounds were sesquiterpene hydrocarbons, which, depending of the harvest time, corresponded to 43-78% of the total essential oil. The other compounds were distributed as monoterpene hydrocarbons, oxygen-containing sesquiterpenes, n-alkanes, 1-alkenes, and oxygen containing monoterpenes, these being a minor group. In H. androsaemum in vitro shoots, sesquiterpene hydrocarbons represented >80% of the respective essential oil. Differences in the essential oil composition were found depending on the harvest time and origin, in vivo versus in vitro, of the plant material. The essential oil sampled in November was characterized by the highest levels of sesquiterpene hydrocarbons and a high number of n-alkanes and 1-alkenes, from C(18) to C(28), whereas that sampled in June of the following year showed the highest levels of n-nonane and 1-octene as well as monoterpene hydrocarbons, the second most representative group. PMID- 12590489 TI - Study of light-induced volatile compounds in goat's milk cheese. AB - Light-induced volatile compounds in goat cheese were studied by a combination of solid phase microextraction (SPME)-gas chromatography (GC)-mass spectrometry (MS), headspace oxygen depletion, and sensory evaluation. Samples stored under fluorescent light for 2 days at 30 degrees C had 90% more volatile compounds and 4 times more headspace oxygen depletion than samples stored in the dark at 30 degrees C. The volatiles 1-heptanol, heptanal, nonanal, and 2-decenal were formed and increased only in the light-stored samples, which may be formed from singlet oxygen oxidation of unsaturated fatty acids. Sensory evaluation showed that samples stored under light had significantly more off-flavor than samples stored in the dark at 30 degrees C (P < 0.05), and 1-heptanol, heptanal, nonanal, and 2 decenal increased the goat cheese off-flavor significantly (P < 0.05). PMID- 12590490 TI - Formation of 5-methyl-4-hydroxy-3[2H]-furanone in cytosolic extracts obtained from Zygosaccharomyces rouxii. AB - Formation of the flavor compound and precursor 4-hydroxy-5-methyl-3[2H]-furanone (HMF, norfuraneol) was demonstrated in cytosolic protein extracts obtained from Zygosaccharomyces rouxii after incubation with a number of carbohydrate phosphates. 4-Hydroxy-5-methyl-3[2H]-furanone was produced from d-fructose-1,6 diphosphate, d-fructose-6-phosphate, d-glucose-6-phosphate, 6-phosphogluconate, d ribose-5-phosphate, and d-ribulose-1,5-diphosphate. Enzyme assays revealed d fructose-1,6-diphosphatase, phosphohexose isomerase, d-glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase activity in the cytosolic extracts. Model studies showed the spontaneous formation of HMF from d-ribulose-5 phosphate. It is assumed that d-ribulose-5-phosphate is generated in cytosolic extracts by the action of the investigated enzymes from the carbohydrate phosphates and is then chemically transformed to HMF. The hypothesis was proven by the production of HMF in solutions containing commercially available enzymes and [6-(13)C]-d-glucose-6-phosphate. PMID- 12590491 TI - Sensory properties of virgin olive oil polyphenols: identification of deacetoxy ligstroside aglycon as a key contributor to pungency. AB - Polyphenols are an important functional minor component of virgin olive oils that are responsible for the key sensory characteristics of bitterness, pungency, and astringency. Polyphenols were isolated from virgin olive oils by using liquid/liquid extraction and then separated by using reverse phase HPLC followed by fraction collection. The sensory qualities of the isolated polyphenols were evaluated, and almost all fractions containing polyphenols were described as bitter and astringent. However, the fraction containing deacetoxy-ligstroside aglycon produced a strong burning pungent sensation at the back of the throat. In contrast, the fraction containing the analogous deacetoxy-oleuropein aglycon, at an equivalent concentration, produced only a slight burning/numbing sensation, which was perceived more on the tongue. No other polyphenol fractions from the analyzed oils produced the intense burning sensation; thus, deacetoxy-ligstroside aglycon is the polyphenol responsible for the majority of the burning pungent sensation found in pungent extra virgin olive oils. PMID- 12590492 TI - Volatile components and aroma active compounds in aqueous essence and fresh pink guava fruit puree (Psidium guajava L.) by GC-MS and multidimensional GC/GC-O. AB - Characterization of the aromatic profile in commercial guava essence and fresh fruit puree by GC-MS yielded a total of 51 components quantified. Commercial essence was characterized to present a volatile profile rich in components with low molecular weight, especially alcohols, esters, and aldehydes, whereas in the fresh fruit puree terpenic hydrocarbons and 3-hydroxy-2-butanone were the most abundant components. In the olfactometric analyses totals of 43 and 48 aroma active components were detected by the panelists in commercial essence and fruit puree, respectively. New components were described for the first time as active aromatic constituents in pink guava fruit (3-penten-2-ol and 2-butenyl acetate). Principal differences between the aroma of the commercial guava essence and the fresh fruit puree could be related to acetic acid, 3-hydroxy-2-butanone, 3-methyl 1-butanol, 2,3-butanediol, 3-methylbutanoic acid, (Z)-3-hexen-1-ol, 6-methyl-5 hepten-2-one, limonene, octanol, ethyl octanoate, 3-phenylpropanol, cinnamyl alcohol, alpha-copaene, and an unknown component. (E)-2-Hexenal seems to be more significant to the aroma of the commercial essence than of the fresh fruit puree. PMID- 12590493 TI - Sinapate dehydrodimers and sinapate-ferulate heterodimers in cereal dietary fiber. AB - Two 8-8-coupled sinapic acid dehydrodimers and at least three sinapate-ferulate heterodimers have been identified as saponification products from different insoluble and soluble cereal grain dietary fibers. The two 8-8-disinapates were authenticated by comparison of their GC retention times and mass spectra with authentic dehydrodimers synthesized from methyl or ethyl sinapate using two different single-electron metal oxidant systems. The highest amounts (481 microg/g) were found in wild rice insoluble dietary fiber. Model reactions showed that it is unlikely that the dehydrodisinapates detected are artifacts formed from free sinapic acid during the saponification procedure. The dehydrodisinapates presumably derive from radical coupling of sinapate-polymer esters in the cell wall; the radical coupling origin is further confirmed by finding 8-8 and 8-5 (and possibly 8-O-4) sinapate-ferulate cross-products. Sinapates therefore appear to have an analogous role to ferulates in cross linking polysaccharides in cereal grains and presumably grass cell walls in general. PMID- 12590494 TI - Impact of whey protein emulsifiers on the oxidative stability of salmon oil-in water emulsions. AB - To obtain a better understanding of how the interfacial region of emulsion droplets influences lipid oxidation, the oxidative stability of salmon oil-in water emulsions stabilized by whey protein isolate (WPI), sweet whey (SW), beta lactoglobulin (beta-Lg), or alpha-lactalbumin (alpha-La) was evaluated. Studies on the influence of pH on lipid oxidation in WPI-stabilized emulsions showed that formation of lipid hydroperoxides and headspace propanal was much lower at pH values below the protein's isoelectric point (pI), at which the emulsion droplets were positively charged, compared to that at pH values above the pI, at which the emulsion droplets were negatively charged. This effect was likely due to the ability of positively charged emulsion droplets to repel cationic iron. In a comparison of lipid oxidation rates of WPI-, SW-, beta-Lg-, and alpha-La stabilized emulsions at pH 3, the oxidative stability was in the order of beta-Lg > or = SW > alpha-La > or = WPI. The result indicated that it was possible to engineer emulsions with greater oxidative stability by using proteins as emulsifier, thereby reducing or eliminating the need for exogenous food antioxidants. PMID- 12590495 TI - Elucidation of the effect of formaldehyde and lipids on frozen stored cod collagen by FT-Raman spectroscopy and differential scanning calorimetry. AB - The effect of frozen storage (-10 and -30 degrees C), formaldehyde, and fish oil on collagen, isolated from cod muscle, was investigated. Salt- and acid-soluble collagen fractions as well as insoluble collagen indicated changes in solubility on frozen storage. Differential scanning calorimetry (DSC) showed a highly cooperative transition at 28.2 degrees C for isolated collagen. Changes in the thermodynamic properties of collagen were observed on frozen storage at -10 degrees C compared with the control at -30 degrees C because of changes in structure. In the presence of formaldehyde, there were no changes in the DSC collagen transition; however, in the presence of fish oil there was an increase in enthalpy and an extra peak was observed at 44.6 degrees C, indicating collagen fish oil interaction. Structural changes resulted in a decrease in the solubility of collagen in salt and acid solution. FT-Raman spectra obtained for collagen at 10 degrees C and -30 degrees C confirmed the alteration of the conformation of collagen not only at -10 degrees C but also in the presence of formaldehyde and fish oil. PMID- 12590496 TI - Molecular determinants of the influence of hydrophilic plasticizers on the mechanical properties of cast wheat gluten films. AB - The influence of a set of hydrophilic plasticizers varying in their chain length (ethyleneglycol and longer molecules) on the tensile strength and elongation at break of cast gluten films was studied. When considered on a molar basis (moles of plasticizer per mole of amino acid), the effect of the different plasticizers depended on their respective molecular weights for plasticizer/amino acid ratios in the range from 0.10 to 0.40. However, above a ratio of 0.40-0.50 mol/mol of amino acid, these differences were abolished and both stress and strain reached a plateau value, with all plasticizers studied. In fact, when a homologous series of molecules was considered, the ability for plasticizer to decrease stress and increase strain was closely related to the number of hydrogen bonds the molecule was able to share with the protein network. Ethyleneglycol's efficiency was, however, lower than expected from its hydrogen-bonding potential; a comparison with other diols demonstrated that this was due to the small size of this molecule. The particular effect of glycerol concentration on the films' mechanical properties suggested that other molecular features of the plasticizer, such as the number and position of hydroxide groups in the molecule, were involved in the plasticization mechanism. PMID- 12590497 TI - Beta-glucosidase from the grape native yeast Debaryomyces vanrijiae: purification, characterization, and its effect on monoterpene content of a Muscat grape juice. AB - Six hundred ten yeast colonies isolated from various vineyards in Chile were screened for the presence of a beta-glucosidase activity as well as the resistance to glucose and ethanol inhibition. Among them, Debaryomyces vanrijiae was found to produce high levels of an extracelular beta-glucosidase which was tolerant to glucose (K(i) = 439 mM) and ethanol inhibitions. The enzyme (designated DV-BG) was purified to apparent homogeneity, respectively, by gel filtration, ion-exchange, and chromatofocusing techniques. Its molecular weight was 100 000, and its pI 3.0, optimum pH, and temperature activities were 5.0 and 40 degrees C, respectively, and had a V(max) of 47.6 micromol min(-)(1) mg(-)(1) and a K(m) of 1.07 mM. The enzyme was active against different beta-d-glucosides including glucosidic flavor precursors. The disaccharidic flavor precursors were not substrates for the enzyme. When added to a Muscat grape juice, the concentration of several monoterpenes increased as the consequence of its hydrolytic activity. PMID- 12590499 TI - Resveratrol content of Palomino fino grapes: influence of vintage and fungal infection. AB - The objective of this study was to determine the influence of certain factors on the resveratrol content of Palomino fino grapes, cultivated in the Jerez-Xerez Sherry area, at the moment of harvest. The results show that the resveratrol content is highly influenced by the climatic conditions prior to the period of maturation of the fruit. On the other hand, the gray mold pressure in the vineyard, a fungal infection caused by Botrytis cinerea, increased the resveratrol contents at the early stages of fungal development. When Botrytis development was extensive, the resveratrol content tended to decrease in the juice but tended to increase in the skin. Physiological stress of the plant leads to increases in the resveratrol content, caused as much by the climatic conditions of the vintage as by biotic factors. In this case resveratrol is present mainly in the glycosylated form. PMID- 12590498 TI - Factors Influencing texture retention of salt-free, acidified, red bell peppers during storage. AB - Red bell peppers were stored in a salt-free, acidified cover solution with sulfite as a microbial preservative. The texture retention of stored peppers was evaluated as a function of pH, acid, calcium, blanch treatment, and growing conditions of the peppers. Field-grown peppers softened at a faster rate than greenhouse-grown peppers and exhibited more variability in their ability to maintain texture during storage. Improved firmness retention of red peppers stored at 30 degrees C was observed when the pH was adjusted to be in the range of 3.4-3.8, when at least 5 mM calcium was added to the peppers, and when red peppers were blanched at 75 degrees C for at least 1 min. PMID- 12590500 TI - Effect of tannins from Quercus suber and Quercus coccifera leaves on ethanol induced gastric lesions in mice. AB - The gastroprotective effects of 70% acetone extracts of Quercus suber and Quercus coccifera leaves and of tannins (pedunculagin, castalagin, phillyraeoidin A, and acutissimin B) purified from these extracts were examined in the mouse using the ethanol-induced gastric ulcer model. Both extracts (25, 50, and 100 mg/kg), given orally, prevented the formation of ethanol-induced lesions in the stomach. The percent protection varied between 68 and 91%. Purified tannins (50 mg/kg) were also effective in protecting the stomach against ethanol, and the percent protection varied from 66 to 83%. Castalagin was the most potent. Both extracts and all of the tannins tested (10, 25, and 50 microg/mL) strongly inhibited (55 65%) the lipid peroxidation of rabbit brain homogenate. These results suggest that the gastroprotective effects of extracts of Q. suber and Q. coccifera leaves and the purified tannins in this experimental model are related to their anti lipoperoxidant properties. PMID- 12590501 TI - Effects of phenolic acids on human phenolsulfotransferases in relation to their antioxidant activity. AB - Sulfate conjugation by phenolsulfotransferase (PST) enzyme is an important process in the detoxification of xenobiotics and endogenous compounds. There are two forms of PST that are specific for the sulfation of small phenols (PST-P) and monoamines (PST-M). Phenoilc acids have been reported to have important biological and pharmacological properties and may have benefits to human health. In the present study, human platelets were used as a model to investigate the influence of 13 phenolic acids on human PST activity and to evaluate the relationship to their antioxidant activity. The results showed that chlorogenic acid, syringic acid, protocatechuic acid, vanillic acid, sinapic acid, and caffeic acid significantly (p < 0.05) inhibited the activities of both forms of PST by 21-30% at a concentration of 6.7 microM. The activity of PST-P was enhanced (p < 0.05) by p-hydroxybenzoic acid, gallic acid, gentisic acid, o coumaric acid, p-coumaric acid, and m-coumaric acid at a concentration of 6.7 microM, whereas the activity of PST-M was enhanced by gentisic acid, gallic acid, p-hydroxybenzoic acid, and ferulic acid. The phenolic acids exhibited antioxidant activity as determined by the oxygen radical absorbance capacity (ORAC) assay and Trolox equivalent antioxidant capacity (TEAC) assay, especially gallic acid, p hydroxybenzoic acid, gentisic acid, and coumaric acid, which had strong activity. The overall effect of phenolic acids tested on the activity of PST-P and PST-M was well correlated to their antioxidant activity of ORAC value (r = 0.71, p < 0.01; and r = 0.66, p < 0.01). These observations suggest that antioxidant phenolic acids might alter sulfate conjugation. PMID- 12590502 TI - Quantitative evaluation of antioxidant components in prunes (Prunus domestica L.). AB - Prunes are known to show high antioxidant activity on the basis of the oxygen radical absorbance capacity (ORAC), and their major antioxidant components are caffeoylquinic acid isomers. The aim of this study is to prove the contribution of caffeoylquinic acid isomers to the ORAC of prunes, and to investigate the existence of other antioxidant components. Caffeoylquinic acid isomers in ethanol (EtOH) extracts of prunes were quantified by HPLC analysis, and the degree of contribution of these isomers to the ORAC was found to be 28.4%; hence, it was speculated that the remaining ORAC is dependent on other antioxidant compounds. EtOH extract was partitioned between hexane and H(2)O. The H(2)O layer was further separated into H(2)O and 2-100% methanol (MeOH) eluates by Diaion HP-20 column chromatography. Both the H(2)O and 50% MeOH eluates showed high values of total phenolics and ORAC, although the contribution of caffeoylquinic acid isomers to the ORAC was low. Therefore, it was predicted that unknown antioxidants exist in these fractions, and several compounds were identified by HPLC analysis. Furthermore, hydrolysis of EtOH extract residue led to higher levels of total phenolics and ORAC, and these results suggested the existence of conjugated antioxidant components in prunes. PMID- 12590503 TI - Determination of the total antioxidant activity of fruits and vegetables by a liposome assay. AB - The effects of mixtures of antioxidants on the oxidation of phospholipids have been investigated in large unilamellar liposomes following initiation by 2,2' azobis(2-aminopropane) dihydrochloride. The lag phase increased linearly with antioxidant concentration. The lag phases of mixtures containing alpha-tocopherol with ascorbic acid showed synergy between the antioxidants, but mixtures of beta carotene with alpha-tocopherol or ascorbic acid were not synergistic. The liposome system was used to investigate the total antioxidant activity of lipid- and water-soluble extracts from 16 samples of fruits, vegetables, and related food products. The water-soluble extracts caused greater increases in lag phase than the lipid-soluble extracts. The lag phase of liposomes containing the water soluble extracts from fruits and vegetables increased linearly with the total phenolic concentration, with the continental salad extract having the longest lag phase. The lipid-soluble extract from apples caused the largest increase in lag phase of the lipid-soluble extracts. The lag phases of the lipid-soluble and water-soluble extracts of all fruits and vegetables studied were additive, but no synergy was detected. The lag phase of the liposomes containing both the water soluble and lipid-soluble extracts varied from 611.5 min for the continental salad extracts to 47.5 min for the cauliflower extracts. PMID- 12590504 TI - Expression and characterization of sweet potato invertase in Pichia pastoris. AB - An invertase cDNA (Ibbetafruct1) was cloned from sweet potato leaves and characterized. The deduced amino acid sequence of the Ibbetafruct1-encoded protein was closely related to vacuolar invertases and included the WECVD catalytic domain characteristic of them. An expression plasmid containing the coding region of Ibbetafruct1 under the control of the alcohol oxidase promoter was used to transform the methylotrophic yeast Pichia pastoris. The biochemical properties for the expressed recombinant enzyme, which was determined to be the acid beta-fructofuranosidase with an acidic pI value (5.1), were similar to those of vacuolar invertases purified from sweet potato. Periodic acid/Schiff staining and Con A-Sepharose gel-binding experiments revealed the recombinant invertase to be a glycoprotein containing glucose and/or mannose residues. Furthermore, the carbohydrate moiety appears to be a key determinant of the enzyme's sucrose hydrolysis activity, substrate affinity, and thermal stability. PMID- 12590505 TI - Buckwheat honey increases serum antioxidant capacity in humans. AB - Honey has been known to exert significant in vitro antioxidant activity, in part due to its phenolic content. However, conclusions that the antioxidants in honey are or are not efficacious in the human body cannot be reached if its antioxidant action is not assessed as part of a human study. In the present study, the acute effect of consumption of 500 mL of water, water with buckwheat honey, black tea, black tea with sugar, or black tea with buckwheat honey on serum oxidative reactions was examined in 25 healthy men. Antioxidant capacity of human serum samples was measured using different methods: the oxygen radical absorbance capacity (ORAC) assay, ex vivo susceptibility of serum lipoprotein to Cu(2+) induced oxidation, and the thiobarbituric acid reactive substances (TBARS) assay. The results showed that the serum antioxidant capacity determined by ORAC increased significantly (p < 0.05) by 7% following consumption of buckwheat honey in water. No significant changes in serum antioxidant capacity could be established after the consumption of any of the other beverages. Ex vivo serum lipoprotein oxidation and TBARS values were not significantly altered after consumption of any of the five beverages. This study provides primary evidence of the in vivo antioxidant activity of buckwheat honey. However, long-term studies and epidemiological data are necessary to investigate whether honey consumption can exert overall antioxidant-related health benefits. PMID- 12590506 TI - Antioxidant properties and phytochemical characteristics of extracts from Lactuca indica. AB - Lactuca indica (Compositae) is an edible wild vegetable, used as a folk medicine in anti-inflammatory, antibacterial, and other medications in Asia. This is the first scientific evaluation of the chemopreventive therapeutic properties of L. indica using five antioxidation assay systems. An extract from L. indica was found to possess significant free radical scavenging activity, effectively protecting phix174 supercoiled DNA against strand cleavage and reducing oxidative stress in human promyelocytic leukemia HL-60 cells. Moreover, extracts of L. indica almost totally inhibited nitric oxide production and the mRNA expression of inducible nitric oxide synthase, at a dosage of 100 microg/mL, in LPS stimulated macrophage RAW264.7 cells. Bioactivity-guided chromatographic fractionation and metabolite profiling coupled with spectroscopic analyses revealed that the six phenolic compounds, that is, protocatechulic acid (1), methyl p-hydroxybenzoate (2), caffeic acid (3), 3,5-dicaffeoylquinic acid (4), luteolin 7-O-beta-glucopyranoside (5), and quercetin 3-O-beta-glucopyranoside (6), are the major antioxidative constituents in the L. indica extract. PMID- 12590507 TI - A [2]catenane constructed around a Ru(diimine)(3)(2+) complex used as a template. AB - A [2]catenane has been constructed using an octahedral complex of the Ru(diimine)( 2+)(3) family as a scaffold. Two diimine chelates have been incorporated in a ring prior to the ruthenium(II) complexation reaction. The macrocyclic complex thus obtained has been subsequently threaded by a long linear fragment containing the third chelate. The rutheniuml(II)-complexed catenane, cyclized by ring-closing metathesis, is the first example of an interlocking ring system built around an octahedral tris-chelate complex. PMID- 12590508 TI - Cryogenic CO oxidation on TiO(2)-supported gold nanoclusters precovered with atomic oxygen. AB - Bulk gold has long been regarded as a noble metal, having very low chemical and catalytic activity. However, metal oxide-supported gold particles, particularly those that are less than 5 nm in diameter, have been found to have remarkable catalytic properties. In this study we show that impinging gas-phase CO molecules react readily with oxygen adatoms preadsorbed on Au/TiO(2)(110) to produce CO(2) even under conditions in which the sample is cryogenically cooled. Gold particle size seems to have little effect on the CO oxidation reaction when oxygen adatoms are preadsorbed. We also show that as the oxygen adatom coverage increases, the rate of CO oxidation decreases on Au/TiO(2) at cryogenic temperatures. PMID- 12590509 TI - Benzylation of aromatic compounds with different crystallites of MgO. AB - Magnesium oxide nanocrystals are shown to be selective catalysts for the benzylation of aromatics with benzyl chloride, in the order xylene > toluene >> benzene. Normal polycrystalline MgO is not as effective as the nanomaterials. However, at the highest surface area, normally most reactive MgO samples (aerogel prepared AP-MgO) were poorer catalysts than a hexagonal platelet form (CP-MgO) of lower surface area. During the catalysis, the MgO is partially converted to MgCl(2), but crystal shape differences are not washed out. Rate data, differential thermal analysis, and consideration of crystal shapes and faces suggest that the CP-MgO form is most catalytically active because of optimal adsorption, molecular trafficking, and desorption energies. The AP-MgO apparently adsorbs the reactants too strongly, and thereby this hinders the catalytic process. These results serve as another example of where nanocrystalline shape plays an important role. PMID- 12590510 TI - (Tl[Au(C(6)Cl(5))(2)])(n): A vapochromic complex. AB - The X-ray structure of (Tl[Au(C(6)Cl(5))(2)])(n), 1, consists of 1-D linear polymer chains parallel to the crystallographic z axis. The crystal structure of 1 has channels that run parallel to these chains with interatom distances in the range 3.231-4.076 A. There are holes in these channels with diameters as large as 10.471 A, which can accommodate a variety of solvents. Complex 1 displays reversible vapochromic emission and absorption spectral behavior when the solid is exposed to a variety of organic vapors such as acetone, acetonitrile, triethylamine, acetylacetone, tetrahydrothiophene, 2-fluoropyridine, tetrahydrofuran, and pyridine. Complex 1 is luminescent at room temperature and at 77 K in the solid state. UV excitation at 495 nm leads to an emission at 531 nm. PMID- 12590511 TI - Binding and activation of aromatic molecules by a molybdenum pi-base. AB - Compounds having the form TpMo(NO)(1-methylimidazole)(eta(2)-L(pi)) (Tp = hydridotris(pyrazolyl)borate; L(pi) = cyclohexene, naphthalene, furan, thiophene, and acetone) were synthesized in 31-41% yield by the reduction of TpMo(NO)Br(2) in the presence of 1-methylimidazole and the respective pi-acidic organic ligand. The structure of the naphthalene complex was confirmed by single-crystal X-ray diffraction. Degradation studies showed the bound aromatics to have half-lives of 37-236 h in acetone solution at 20 degrees C. A tandem addition across the bound ring of the naphthalene complex yielded a free 1,2-dihydronaphthalene in 80% yield following decomplexation. PMID- 12590512 TI - A highly reduced v(2)(3+) unit with a metal-metal bond order of 3.5. AB - We report the isolation and structural characterization of the first four-bladed paddlewheel complex having an M(2)(3+) core in K(THF)(3)[V(2)(DPhF)(4)], where DPhF is the anion of N,N'-diphenylformamidine. This dark green, paramagnetic compound was obtained by reduction of diamagnetic, triply bonded V(2)(DPhF)(4) with KC(8). The V-V bond distance in K(THF)(3)[V(2)(DPhF)(4)] is 1.9295(8) A, 0.05 A shorter than that found in the V(2)(4+) species. The shortening of the bond, combined with a 15-line hyperfine splitting pattern in the EPR spectrum of K(THF)(3)[V(2)(DPhF)(4)], indicates that the unpaired electron is delocalized over the two vanadium atoms, giving rise to a bond order of 3.5. PMID- 12590513 TI - Novel iminium ion equivalents prepared through C-H oxidation for the stereocontrolled synthesis of functionalized propargylic amine derivatives. AB - Access to stereochemically complex, polyfunctionalized amine derivatives is made possible using novel oxathiazinane N,O-acetal starting materials. These heterocycles are prepared through intramolecular sulfamate ester C-H insertion with a Rh(2+)-carboxylate catalyst and PhI(OAc)(2) as the terminal oxidant. Such compounds function as unique iminium ion equivalents to which nucleophilic alkynylzinc reagents add smoothly in the presence of BF(3).OEt(2). The coupled products are isolated in high yield (63-92%) and with good levels of diastereoinduction (6 --> 20:1). The alkyne-substituted oxathiazinanes serve as versatile building blocks and may be further manipulated through nucleophilic ring-opening reactions of the sulfamate core. The efficient construction of 1,7,8 trihydroxyindolizidine in six steps and in 34% overall yield highlights the power of these combined methods for synthesis. PMID- 12590514 TI - One-electron oxidation of the guanine moiety of 2'-deoxyguanosine: influence of 8 oxo-7,8-dihydro-2'-deoxyguanosine. AB - The influence of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) on riboflavin and UVA-mediated one-electron oxidation of an aqueous aerated solution of 2' deoxyguanosine (dGuo) has been studied. Using labeled experiments, we have demonstrated that, despite not being able to detect significant amounts of 8 oxodGuo upon one-electron oxidation of dGuo, 8-oxodGuo is indeed produced but is further rapidly degraded to oxidized nucleosides. Evidence is provided showing that an efficient electron transfer reaction from 8-oxodGuo to the guanine radical cation or rather its deprotonated form occurs, giving rise to the specific decomposition of 8-oxodGuo together with the restitution of dGuo. It could be concluded that 8-oxodGuo efficiently protects dGuo from decomposition by the one-electron oxidation reaction. PMID- 12590515 TI - Thermodynamic coupling of the loop and stem in unusually stable DNA hairpins closed by CG base pairs. AB - For certain DNA hairpin loops, a CG closing base pair has enhanced stability over other closing base pairs, which cannot be explained by the current nearest neighbor model. We report the use of three-carbon (C3) spacers to investigate the expandability of DNA hairpin loops and the coupling between the loop and closing base pair. Inserting the C3-spacers at most positions in these model loops produced only a modest stabilization or destabilization except for insertion between the 5' end of the loop and the CG closing base pair, which gave a large destabilization. Further investigation on tetraloops and triloops with other closing base pairs established that this destabilization is specific to the unusually stable CG closing base pair. Studies with the nucleotide analogues 2 aminopurine and 2,6-diaminopurine indicated that this stabilization may be due to coupling between functional groups on the first base of the loop and the CG closing base pair. The C3-spacers provide a simple way to interrupt potential interactions and thereby probe loop/stem coupling. PMID- 12590516 TI - Modulation of substrate binding to naphthalene 1,2-dioxygenase by rieske cluster reduction/oxidation. AB - The active site of the oxygenase component of naphthalene 1,2-dioxygenase (NDO) contains a Rieske Fe-S cluster and a mononuclear non-heme iron, which are contributed by different alpha-subunits in the (alphabeta)(3) structure. The enzyme catalyzes cis-dihydroxylation of aromatic substrates in addition to numerous other adventitious oxidation reactions. High-resolution Mims (2)H-ENDOR spectra have been recorded for the NO-ferrous center of NDO bound with d(8) naphthalene and d(2)-naphthalene; spectra were collected for the enzyme with the Rieske diiron center both in its oxidized and in its reduced states. A sharp quartet ENDOR pattern from a nearby deuteron of substrate was detected for each substrate. Examination of the sample prepared with 1,4-dideutero-naphthalene shows that the signal arises from D1. The ENDOR data place D1 at a distance of ca. 4.4 A from the mononuclear Fe and with the Fe-D vector being roughly along the Fe-N(O) direction. Because reduction of the Rieske cluster is required for O(2) binding and subsequent catalysis, the effect of its oxidation state on substrate binding was examined. The spectra from the NDO-naphthalene complex reveal two different binding conformations, which change in relative population when the oxidation state of the Rieske cluster is changed. This shift, and the conformational coupling it implies, may hold the key to both oxygen gating and oxygen reactivity for Rieske aromatic dioxygenases. PMID- 12590517 TI - Native conformational tendencies in unfolded polypeptides: development of a novel method to assess native conformational tendencies in the reduced forms of multiple disulfide-bonded proteins. AB - Oxidative folding is the concomitant formation of the native disulfide bonds and the native tertiary structure from the reduced and unfolded polypeptide. Of interest is the inherent conformational tendency (bias) present in the reduced polypeptide to dictate the formation of the full set of native disulfide bonds. Here, by application of a novel tool, we have been able to assess this "native conformational tendency" present in reduced and unfolded bovine pancreatic ribonuclease A (RNase A). The essence of this method lies in the ability of the oxidant [Pt(en)(2)Cl(2)](2+) (where "en" is ethylenediamine) to oxidize disulfide bonds under conditions in which both reduction and disulfide reshuffling, which are essential for rearranging non-native disulfide bonds, are extremely slow. When applied to RNase A, the method revealed little or no bias toward formation of the full native set of disulfide bonds in the fully reduced protein. PMID- 12590518 TI - Synthesis of 1,2-diheteroatom-substituted alkenes via rhodium-catalyzed intramolecular hydrogen transfer. AB - While 1,2-diheteroatom-substituted alkenes represent useful synthetic building blocks, there are no simple, general procedures available for preparation of such compounds. We describe here a simple catalytic procedure based on rhodium C-H bond activation for the synthesis of diheteroatom alkenes of the type R(3)SiOCH=CHG (G = -OR, -NRR'). PMID- 12590519 TI - Porphyrin deposition induced by UV irradiation. AB - Simple irradiation by UV light of porphyrin TpyP and its zinc(II) and manganese(III) metal derivatives in chlorinated solvents leads to the deposition on silica of mesoscale aggregates. The process is due to the photochemical decomposition of the solvent with generation of HCl, which causes the precipitation of a protonated form. The morphology of the resulting objects is driven by the nature of the inserted metal ion. PMID- 12590520 TI - Thiyl radicals abstract hydrogen atoms from the (alpha)C-H bonds in model peptides: absolute rate constants and effect of amino acid structure. AB - Thiyl radicals are important intermediates in biological oxidative stress and enzymatic reactions, for example, the ribonucleotide reductases. On the basis of the homolytic bond dissociation energies (BDEs) only, the (alpha)C-H bonds of peptides and proteins would present suitable targets for hydrogen abstraction by thiyl radicals. However, additional parameters such as polar and conformational effects may control such hydrogen-transfer processes. To evaluate the potential of thiyl radicals for hydrogen abstraction from (alpha)C-H bonds, we provide the first absolute rate constants for these reactions with model peptides. Thiyl radicals react with (alpha)C-H bonds with rate constants between 1.7 x 10(3) M( 1) s(-1) (N-acetylproline amide) and 4 x 10(5) M(-1) s(-1) (sarcosine anhydride). However, the correlation of rate constants with BDEs is poor. Rather, these reactions may be controlled by conformation and dynamic flexibility around the (alpha)C-H bonds. PMID- 12590521 TI - Shishijimicins A-C, novel enediyne antitumor antibiotics from the ascidian Didemnum proliferum(1). AB - Three new compounds of the enediyne antibiotics, shishijimicins A-C (1-3), have been isolated from the ascidian Didemnum proliferum. They encompass a novel sugar component, which is a conjugation product of a hexose and a beta-carboline, attached to the calicheamicinone aglycone. Their structures have been determined by interpretation of spectral data. Shishijimicins showed extremely potent cytotoxicity against HeLa cells with IC(50) values of 1.8-6.9 pM. PMID- 12590522 TI - Torsional barriers for planar versus twisted singlet styrenes. AB - Kinetic modeling of the temperature-dependent lifetimes of styrene and several methyl-substituted styrenes has been used to obtain the torsional barriers for singlet-state C=C rotation. The barrier for C=C torsion is found to be correlated with the ground-state phenyl-vinyl dihedral angle, varphi, planar styrenes having barriers of approximately 6.5 kcal/mol and moderately twisted styrenes having smaller barriers. Highly twisted styrenes undergo exceptionally rapid intersystem crossing. This unexpected dependence of excited-state behavior on varphi is attributed to a change in the character of the excited states from delocalized for planar styrenes to localized for highly twisted styrenes. PMID- 12590523 TI - Self-assembly of spherical colloids into helical chains with well-controlled handedness. AB - Physical constraint and capillary force have been combined to provide a generic approach to assemble achiral building blocks such as monodisperse spherical colloids into helical mesostructures. The key component of this process is an array of V-shaped grooves anisotropically etched in the surface of a Si(100) wafer. The structural arrangement among the spherical colloids is determined by the ratio between the width of the V-grooves and the diameter of the colloids. Double-layered structures with a helical morphology will be formed when this ratio falls between 2.70 and 2.85. The exact handedness of these helical structures could be controlled by varying the relative orientation of capillary force with respect to the longitudinal axis of the spirals. The processing of an achiral material into helical mesostructures having well-controlled handedness should allow us to explore new properties that this material otherwise does not exhibit. The self-assembly process may also provide valuable insights to improve our understanding on the mechanism(s) by which pure enantiomers with a particular sense of rotation evolved in nature. PMID- 12590524 TI - Single-precursor, one-pot versatile synthesis under near ambient conditions of tunable, single and dual band fluorescing metal sulfide nanoparticles. AB - We present a simple and versatile method for the synthesis of high-quality size controlled metal sulfide nanoparticles. A single compound (metal xanthate) is the precursor. A Lewis-base solvent is used to achieve a low reaction temperature of 50-150 degrees C, usually in air. Demonstrated with CdS, the precise control over the particle size (by regulating the temperature or the concentration) enables tuning the absorption and emission spectra of the particles. We also can control the relative intensity of the narrow (30-35 nm wide) excitonic emission (tunable in the range 430-480 nm with approximately 2% fluorescence quantum efficiency) and the broad emission associated with deep surface traps (in the range 550-700 nm). Using the same precursor CdS/ZnS core/shell particles are produced with a high PL yield ( approximately 14%). PMID- 12590525 TI - Copper ion-induced activation and asymmetric benzoylation of 1,2-diols: kinetic chiral molecular recognition. AB - New catalytic ability of copper(II) ion has been exploited for monobenzoylation of 1,2-diols. The catalyst can be readily modified by ligation to acquire higher stereoselectivity. Highly effective kinetic resolution of dl-1,2-diols was achieved. The enantiodiscrimination process was clearly shown to be controlled by the kinetics of acylation of dl-1,2-diols. The catalytic method was successfully applied to asymmetric desymmetrization of meso-hydrobenzoin. PMID- 12590526 TI - Designer chiral quaternary ammonium bifluorides as an efficient catalyst for asymmetric nitroaldol reaction of silyl nitronates with aromatic aldehydes. AB - Designer chiral quaternary ammonium bifluoride 1 has been prepared, and both its catalytic and its chiral efficiency have been clearly demonstrated by achieving the first catalytic asymmetric nitroaldol reaction of silyl nitronate with aldehydes. For instance, the reaction of trimethylsilyl nitronate 2 (R(1) = Me) with benzaldehyde (R(2) = Ph) in THF in the presence of (S,S)-1 (2 mol %) proceeded smoothly at -78 degrees C, giving the corresponding nitroaldol adduct 3 (R(1) = Me, R(2) = Ph) in 92% isolated yield (anti/syn = 92:8) with 95% ee (anti isomer). The method was found to be successfully applicable to other aromatic aldehydes and silyl nitronates, and a high level of anti selectivity and enantiomeric excess was constantly observed. This finding should lead to the further development of fluoride ion-catalyzed asymmetric carbon-carbon bond forming reactions. PMID- 12590527 TI - Mechanism of the palladium-catalyzed intramolecular hydroalkylation of 7-octene 2,4-dione. AB - Cyclization of (E)-7,8-dideuterio-7-octene-2,4-dione [(E)-1-7,8-d(2)] catalyzed by PdCl(2)(CH(3)CN)(2) (2) formed cis-2-acyl-3,4-dideuteriocyclohexanone (cis-3 3,4-d(2)) in 64% yield as the exclusive isotopomer. This experiment, in conjunction with additional deuterium labeling experiments, was in accord with a mechanism for the conversion of 1 to 3 catalyzed by 2 involving attack of the enol carbon atom on a palladium-complexed olefin followed by palladium migration and protonolysis from a palladium enolate complex. PMID- 12590528 TI - Asymmetric organocatalysis of 4 + 3 cycloaddition reactions. AB - Treatment of several 4-trialkylsilyloxypentadienals with a chiral secondary amine, trifluoroacetic acid, and a diene resulted in the formation of 4 + 3 cycloaddition products, which were enantiomerically enriched. This represents the first examples of the asymmetric organocatalysis of the 4 + 3 cycloaddition reaction. 2,5-Dimethylfuran reacted with 4-trimethylsilyloxy-2,4-pentadienal in the presence of catalytic amounts of chiral amine and acid to afford a cycloadduct in 64% yield as a single diastereomer with an enantiomeric excess of 89%. PMID- 12590529 TI - Direct test of the Gaussian-chain model for treating residual charge-charge interactions in the unfolded state of proteins. AB - The Gaussian-chain model for treating residual charge-charge interactions was critically tested by recent experimental pK(a) results for individual Asp, Glu, and His residues in the unfolded drkN SH3 domain. Predicted pK(a)'s were in good agreement with experiment. The clustering of Asp and Glu residues along the sequence was suggested to limit pK(a) shifts and contribute to the folding stability by destabilizing the unfolded state. PMID- 12590530 TI - Functionalized boron nitride nanotubes with a stannic oxide coating: a novel chemical route to full coverage. AB - BN nanotubes are coated with SnO(2) by a simple chemical reaction in solution. BN nanotubes are stirred inside a SnCl(2) solution at room temperature for 1 h. The coating is uniform with complete coverage. The coating thickness is typically 1-5 nm. The coating layer contains SnO(2) nanoparticles with sizes ranging from 1 to 5 nm. PMID- 12590531 TI - Synthetic, structural and binding studies of the 4,6-dimethyldibenzothiophene complex [(eta(5)-C(5)Me(5))Ru(CO)(2)(eta(1)(S)-4,6-Me(2)DBT)]BF(4): toward an understanding of deep hydrodesulfurization. AB - The synthesis of the first completely characterized transition-metal complex containing a sulfur-bound 4,6-dimethyldibenzothiophene (4,6-Me(2)DBT) ligand, [CpRu(CO)(2)(eta(1)(S)-4,6-Me(2)DBT)]BF(4) (1) (Cp = eta(5)-C(5)Me(5)), is reported. X-ray studies of 1 and its 4-methyldibenzothiophene and dibenzothiophene analogues, [CpRu(CO)(2)(eta(1)(S)-4-MeDBT)]BF(4) (2) and [CpRu(CO)(2)(eta(1)(S)-DBT)]BF(4) (3), show that the Ru-S bond distances increase in the order, 3 < 2 < 1. Equilibrium studies on the series of [CpRu(CO)(2)(eta(1)(S)-DBTh)](+) compounds, where DBTh = DBT, 4-MeDBT, 4,6 Me(2)DBT, and 2,8-Me(2)DBT, show that the relative binding strengths of the dibenzothiophene ligands increase in the order 4,6-Me(2)DBT (1) < 4-MeDBT (20.2(1)) < DBT (62.7(6)) < 2,8-Me(2)DBT (223(3)). These results are the first to quantify the steric effect of 4- and 6-methyl groups on the sulfur-coordinating ability of dibenzothiophenes to transition-metal centers. They are also consistent with the proposal that 4- and 6-methyl groups reduce the coordination of dibenzothiophenes to active metal sites on hydrodesulfurization catalysts, which could account for the slow rate of 4-MeDBT and 4,6-Me(2)DBT hydrodesulfurization in petroleum feedstocks. PMID- 12590532 TI - Synthesis of stable derivatives of c(62): the first nonclassical fullerene incorporating a four-membered ring. AB - A rational synthetic approach to the first four-membered ring-containing derivatives of C(62) is reported. They were synthesized by an inverse electron demand Diels-Alder reaction of 3,6-diaryl-1,2,4,5-tetrazines with C(60) in o dichlorobenzene, followed by visible light irradiation at reflux. The structure of these nonclassical fullerenes derivatives was determined by X-ray single crystal diffraction. PMID- 12590533 TI - Ring-opening reaction of phosphorus-bridged [1]ferrocenophane via ring slippage from eta(5)- to eta(1)-Cp. AB - A reaction mechanism was investigated for a ring-opening reaction of RP(E) bridged [1]ferrocenophane, where RP(E) = PhP(S) (3a), PhP (3b), and MesP (3c) (Mes = 2,4,6-trimethylphenyl). Irradiation of UV-vis light in the presence of an excess amount of P(OMe)(3) transformed 3a to [Fe(PhP(S)(eta(5)-C(5)H(4))(eta(1) C(5)H(4)))(P(OMe)(3))(2)] (4a), in which one of the two cyclopentadienyl (Cp) rings of 3a changed its coordination mode from eta(5) to eta(1) and vacant coordination sites thus formed on the iron center were occupied by two P(OMe)(3) ligands. The molecular structure of 4a was determined by X-ray analysis, in which eta(1)-Cp adopted a 1-Fe-2-P-1,3-cyclopentadiene structure. Under the same reaction conditions, 3b and 3c also gave similar ring-slipped products 4b and 4c, respectively. Photolysis of 3a using more strongly coordinating PMe(3) in place of P(OMe)(3) led to complete dissociation of a Cp ligand from the iron center to form [Fe(PhP(S)(eta(5)-C(5)H(4))(C(5)H(4)))(PMe(3))(3)] (5). The formation of the ring-slipped and -dissociated products on the photolysis of 3 strongly supports the view that photolytic ring-opening polymerization of 3 proceeds via an unprecedented Fe-Cp bond cleavage mechanism. PMID- 12590534 TI - Unusual coordination modes of arylthiolates in Mo(eta(5)-SC(6)H(3)-2,6 (SiMe(3))(2))(eta(7)-SC(6)H(3)-2,6-(SiMe(3))(2)). AB - Treatment of MoCl(3)(thf)(3) with LiSC(6)H(3)-2,6-(SiMe(3))(2) (LiSAr) resulted in formation of the pi-sandwiched bis-arylthiolato complex, Mo(eta(5)-SC(6)H(3) 2,6-(SiMe(3))(2))(eta(7)-SC(6)H(3)-2,6-(SiMe(3))(2)) (1), while the analogous reaction with LiSC(6)H(3)-2-Ph-6-SiMe(3) afforded the trithiolate complex Mo(SC(6)H(3)-2-Ph-6-SiMe(3))(3) (3). The acetonitrile adduct Mo(SAr)(2)(CH(3)CN)(3) (2) was isolated from the CH(3)CN solution of 1, in which one acetonitrile is coordinated to the metal center in an eta(2)-fashion. Structures of 1, 2, and 3 have been determined by X-ray diffraction. PMID- 12590535 TI - Intercalation of trioxatriangulenium ion in DNA: binding, electron transfer, x ray crystallography, and electronic structure. AB - Trioxatriangulenium ion (TOTA(+)) is a flat, somewhat hydrophobic compound that has a low-energy unoccupied molecular orbital. It binds to duplex DNA by intercalation with a preference for G-C base pairs. Irradiation of intercalated TOTA(+) causes charge (radical cation) injection that results in strand cleavage (after piperidine treatment) primarily at GG steps. The X-ray crystal structure of TOTA(+) intercalated in the hexameric duplex d[CGATCG](2) described here reveals that intercalation of TOTA(+) results in an unusually large extension of the helical rise of the DNA and that the orientation of TOTA(+) is sensitive to hydrogen-bonding interactions with backbone atoms of the DNA. Electronic structure calculations reveal no meaningful charge transfer from DNA to TOTA(+) because the lowest unoccupied molecular orbital of TOTA(+), (LUMO)(T), falls in the gap between the highest occupied molecular orbital, (HOMO)(D), and the (LUMO)(D) of the DNA bases. These calculations reveal the importance of backbone, water, and counterion interactions, which shift the energy levels of the bases and the intercalated TOTA(+) orbitals significantly. The calculations also show that the inserted TOTA(+) strongly polarizes the intercalation cavity where a sheet of excess electron density surrounds the TOTA(+). PMID- 12590536 TI - Design of triple helix forming C-glycoside molecules. AB - The modeling, synthesis, and characterization of oligomers containing 2 aminoquinazolin-5-yl 2'-deoxynucleotide residues are reported. The 2 aminoquinazoline residues sequence specifically bind via Hoogsteen base pairing as a third strand in the center of the major groove at T:A base pair Watson-Crick duplex sequences. Evidence for the formation of a sequence specific three stranded structure is based on thermal denaturation UV-vis and fluorescence studies. The novel 2-aminoquinazoline C-nucleotide is a component of a system designed to overcome the homopurine requirement for triple helix structures. PMID- 12590537 TI - Tandem electrostatic effect from the first to the third aglycon in the trimeric RNA owing to the nearest-neighbor interaction. AB - We here show an electrostatic polar-pi interaction from the first to the third aglycon, via the second aglycon, in the ground state in two single stranded trimeric RNAs, 5'-GpA(1)pA(2)-3' (3) and 5'-GpApC-3' (4), as a result of intramolecular nearest neighbor offset-stacking. The experimental evidence in support of this conclusion has been obtained by comparing the pK(a)s of each aglycone in the two trimers with those of guanosine 3'-ethyl phosphate, GpEt (1) and 5'-GpA-3' (2): Thus, the pK(a) of N(1)-H of guanin-9-yl of 5'-GpA(1)pA(2)-3' (3) could be measured by pH titration (pH 7.3-11.6) of its own deltaH8G (pK(a) 9.75 +/- 0.02) as well as from deltaH8A(1) (pK(a) 9.72 +/- 0.02) and deltaH2A(1) (pK(a) 9.83 +/- 0.04) of the neighboring pA(1)p moiety and the deltaH8A(2) (pK(a) 9.83 +/- 0.02) of the terminal pA(2) moiety. Similarly, the pH titration of GpApC (4) shows the pK(a) of N(1)-H of guanin-9-yl from its own deltaH8G (pK(a) 9.88 +/ 0.03) as well as from deltaH8A (pK(a) 9.87 +/- 0.01) of the neighboring pAp moiety, and deltaH5/H6C (pK(a) 9.88 +/- 0.01 and 9.90 +/- 0.01 respectively) of the 3'-terminal cytosin-1-yl. This intramolecular nearest neighbor electrostatic interaction in the single-stranded RNA modulates the pseudoaromaticity of the nearest neighbors by almost total transmission of because they constitute an extended array of offset-stacked coupled aromatic heterocycles within a polyanionic sugar-phosphate backbone at the ground state. The enhanced basicity of Gp residue by ca. 0.6 pK(a) unit in the trimers compared to that of the dimer is a result of the change in the electrostatic microenvironment owing to the nearest neighbors in the former (the nucleobases as well as the phosphates). Thus, the from the 5'-guanylate ion to the 3'-end aglycon via the central adenin 9-yl is 55 to 56 kJ mol(-)(1) in each step through a distance spanning approximately 6.8 A in an unfolded state. As a result, the pK(a) of guanin-9-yl moiety has become 9.25 +/- 0.02 in GpEt (1), 9.17 +/- 0.02 in GpA (2), 9.75 +/- 0.02 in GpApA (3), and 9.88 +/- 0.03 in GpApC (4). This means that guanin-9-yl moiety of trimers 3 and 4 is more basic than in the monomer or the dimer. The net outcome of this electrostatic cross-talk between the two neighboring heterocycles is creation of new hybrid aglycones in an oligo or polynucleotide, whose physicochemical property and the pseudoaromatic character are completely dependent both upon the nearest neighbors, and whether they are stacked or unstacked. Thus, this tunable physicochemical property of an aglycon (an array of the extended genetic code) may have considerable implication in our understanding of the specific ligand binding ability of an aptamer, the pK(a) and the hydrogen bonding ability of nucleic acids in a microenvironment, or in the triplet usage by the anticodon-codon interaction in the protein biosynthesis in the ribosome. PMID- 12590538 TI - Active-site structure and electron-transfer reactivity of plastocyanins. AB - The active-site structures of Cu(II) plastocyanins (PCu's) from a higher plant (parsley), a seedless vascular plant (fern, Dryopteris crassirhizoma), a green alga (Ulva pertusa), and cyanobacteria (Anabaena variabilis and Synechococcus) have been investigated by paramagnetic (1)H NMR spectroscopy. In all cases the spectra are similar, indicating that the structures of the cupric sites, and the spin density distributions onto the ligands, do not differ greatly between the proteins. The active-site structure of PCu has remained unaltered during the evolutionary process. The electron transfer (et) reactivity of these PCu's is compared utilizing the electron self-exchange (ESE) reaction. At moderate ionic strength (0.10 M) the ESE rate constant is dictated by the distribution of charged amino acid residues on the surface of the PCu's. Most higher plant and the seedless vascular plant PCu's, which have a large number of acidic residues close to the hydrophobic patch surrounding the exposed His87 ligand (the proposed recognition patch for the self-exchange process), have ESE rate constants of approximately 10(3) M(-)(1) s(-)(1). Removal of some of these acidic residues, as in the parsley and green algal PCu's, results in more favorable protein-protein association and an ESE rate constant of approximately 10(4) M(-)(1) s(-)(1). Complete removal of the acidic patch, as in the cyanobacterial PCu's, leads to ESE rate constants of approximately 10(5)-10(6) M(-)(1) s(-)(1). The ESE rate constants of the PCu's with an acidic patch also tend toward approximately 10(5) 10(6) M(-)(1) s(-)(1) at higher ionic strength, thus indicating that once the influence of charged residues has been minimized the et capabilities of the PCu's are comparable. The cytochromes and Fe-S proteins, two other classes of redox metalloproteins, also possess ESE rate constants of approximately 10(5)-10(6) M( )(1) s(-)(1) at high ionic strength. The effect of the protonation of the His87 ligand in PCu(I) on the ESE reactivity has been investigated. When the influence of the acidic patch is minimized, the ESE rate constant decreases at high [H(+)]. PMID- 12590539 TI - Biomimetic aryl hydroxylation derived from alkyl hydroperoxide at a nonheme iron center. Evidence for an Fe(IV)=O oxidant. AB - Many nonheme iron-dependent enzymes activate dioxygen to catalyze hydroxylations of arene substrates. Key features of this chemistry have been developed from complexes of a family of tetradentate tripodal ligands obtained by modification of tris(2-pyridylmethyl)amine (TPA) with single alpha-arene substituents. These included the following: -C(6)H(5) (i.e., 6-PhTPA), L(1); -o-C(6)H(4)D, o-d(1) L(1); -C(6)D(5), d(5)-L(1); -m-C(6)H(4)NO(2), L(2); -m-C(6)H(4)CF(3), L(3); -m C(6)H(4)Cl, L(4); -m-C(6)H(4)CH(3), L(5); -m-C(6)H(4)OCH(3), L(6); -p C(6)H(4)OCH(3), L(7). Additionally, the corresponding ligand with one alpha phenyl and two alpha-methyl substituents (6,6-Me(2)-6-PhTPA, L(8)) was also synthesized. Complexes of the formulas [(L(1))Fe(II)(NCCH(3))(2)](ClO(4))(2), [(L(n)())Fe(II)(OTf)(2)] (n = 1-7, OTf = (-)O(3)SCF(3)), and [(L(8))Fe(II)(OTf)(2)](2) were obtained and characterized by (1)H NMR and UV visible spectroscopies and by X-ray diffraction in the cases of [(L(1))Fe(II)(NCCH(3))(2)](ClO(4))(2), [(L(6))Fe(II)(OTf)(2)], and [(L(8))Fe(II)(OTf)(2)](2). The complexes react with tert-butyl hydroperoxide ((t)()BuOOH) in CH(3)CN solutions to give iron(III) complexes of ortho hydroxylated ligands. The product complex derived from L(1) was identified as the solvated monomeric complex [(L(1)O(-))Fe(III)](2+) in equilibrium with its oxo bridged dimer [(L(1)O(-))(2)Fe(III)(2)(mu(2)-O)](2+), which was characterized by X-ray crystallography as the BPh(4)(-) salt. The L(8) product was also an oxo bridged dimer, [(L(8)O(-))(2)Fe(III)(2)(mu(2)-O)](2+). Transient intermediates were observed at low temperature by UV-visible spectroscopy, and these were characterized as iron(III) alkylperoxo complexes by resonance Raman and EPR spectroscopies for L(1) and L(8). [(L(1))Fe(II)(OTf)(2)] gave rise to a mixture of high-spin (S = 5/2) and low-spin (S = 1/2) Fe(III)-OOR isomers in acetonitrile, whereas both [(L(1))Fe(OTf)(2)] in CH(2)Cl(2) and [(L(8))Fe(OTf)(2)](2) in acetonitrile afforded only high-spin intermediates. The L(1) and L(8) intermediates both decomposed to form respective phenolate complexes, but their reaction times differed by 3 orders of magnitude. In the case of L(1), (18)O isotope labeling indicated that the phenolate oxygen is derived from the terminal peroxide oxygen via a species that can undergo partial exchange with exogenous water. The iron(III) alkylperoxo intermediate is proposed to undergo homolytic O-O bond cleavage to yield an oxoiron(IV) species as an unobserved reactive intermediate in the hydroxylation of the pendant alpha-aryl substituents. The putative homolytic chemistry was confirmed by using 2-methyl-1 phenyl-2-propyl hydroperoxide (MPPH) as a probe, and the products obtained in the presence and in the absence of air were consistent with formation of alkoxy radical (RO(*)). Moreover, when one ortho position was labeled with deuterium, no selectivity was observed between hydroxylation of the deuterated and normal isotopomeric ortho sites, but a significant 1,2-deuterium shift ("NIH shift") occurred. These results provide strong mechanistic evidence for a metal-centered electrophilic oxidant, presumably an oxoiron(IV) complex, in these arene hydroxylations and support participation of such a species in the mechanisms of the nonheme iron- and pterin-dependent aryl amino acid hydroxylases. PMID- 12590540 TI - Efficient synthesis of NK(1) receptor antagonist aprepitant using a crystallization-induced diastereoselective transformation. AB - An efficient stereoselective synthesis of the orally active NK(1) receptor antagonist Aprepitant is described. A direct condensation of N-benzyl ethanolamine with glyoxylic acid yielded a 2-hydroxy-1,4-oxazin-3-one which was activated as the corresponding trifluoroacetate. A Lewis acid mediated coupling with enantiopure (R)-1-(3,5-bis(trifluoromethyl)phenyl)ethan-1-ol afforded a 1:1 mixture of acetal diastereomers which was converted into a single isomer via a novel crystallization-induced asymmetric transformation. The resulting 1,4-oxazin 3-one was converted via a unique and highly stereoselective one-pot process to the desired alpha-(fluorophenyl)morpholine derivative. Interesting and unexpected [1,2]-Wittig and [1,3]-sigmatropic rearrangements were identified during the optimization of these key steps. In the final step, a triazolinone side chain was appended to the morpholine core. The targeted clinical candidate was thus obtained in 55% overall yield over the longest linear sequence. PMID- 12590541 TI - Stable mechanistically-relevant aromatic-based carbenium ions in zeolite catalysts. AB - The existence of carbenium ion species is assumed in many zeolite catalysis mechanisms. Using computational techniques that include environmental effects, a benzenium-type carbenium ion is identified in zeolite catalysts for the first time. Localization of nearby transition states indicate that this species may play an important role as an intermediate in the bimolecular m-xylene disproportionation reaction. The barrier to back-donation of the proton from the benzenium ion is at least 50 kJ/mol, meaning that this species may be spectroscopically observable. An additional carbenium ion intermediate, formed by abstraction of a hydride from m-xylene, is also predicted. The stability of this second new carbenium ion suggests that aromatic-based carbenium ions are likely to be intermediates in many zeolite-catalyzed reactions. Two types of fundamentally different fully periodic calculations support the stability predictions. PMID- 12590542 TI - Are the enolates of amides and esters stabilized by electrostatics? AB - The fact that amides and esters form less stable enolates than ketones might be seen as evidence that electrostatic stabilization is unimportant in these anions. However, ab initio molecular orbital calculations show that electrostatic stabilization does in fact lie beneath the competing resonance effect that causes the decrease in acidity. The electrostatic contribution is revealed by examining torsionally twisted amide and ester structures in which the pi resonance interactions are largely inhibited. These twisted amides and esters have greater enolate acidity than the corresponding ketones. Qualitatively similar behavior is observed with respect to protonation, such that twisted amides and esters are generally less basic than the reference ketones, in striking contrast to their behavior in the normal geometries. PMID- 12590543 TI - Thermochemistry of arylselanyl radicals and the pertinent ions in acetonitrile. AB - Reduction and oxidation potentials of a series of parasubstituted phenylselanyl radicals, XC(6)H(4)Se(*), have been measured using photomodulated voltammetry in acetonitrile. The thermodynamic significance of these data was substantiated through a study of the oxidation process of the pertinent selenolates in linear sweep voltammetry. Both the reduction and the oxidation potentials correlate linearly with the Hammett substituent coefficients sigma and sigma(+) leading in the latter case to slopes, rho(+), of 2.5 and 3.8, respectively. Through comparison of these slopes with those published previously for the O- and S centered analogues, it is revealed that the pi-interaction becomes progressively smaller as the size of the radical center increases in the order O, S, and Se. Solvation energies of the pertinent selenolates and selanylium ions have been extracted from thermochemical cycles incorporating the measured electrode potentials for XC(6)H(4)Se(*) as well as electron affinities and ionization potentials obtained from theoretical calculations at the B3LYP/6-31+G(d) level. The extracted data show the expected overall substituent dependency for both kinds of ions; that is, the absolute value of the solvation energy decreases as the charge becomes more delocalized. The data have also been compared with solvation energies computed using the polarizable continuum model (PCM). Interestingly, we find that, while the model seems to work well for selenolates, it underestimates the solvation of selanylium ions in acetonitrile by as much as 25 kcal mol(-)(1). These large deviations are ascribed to the fact that the PCM method does not take specific solvent effects into account as it treats the solvent as a continuum described solely by its dielectric constant. Gas-phase calculations show that the arylselanylium ions can coordinate covalently to one or two molecules of acetonitrile in strong Ritter-type adducts. When this strong interaction is included in the solvation energy calculations by means of a combined supermolecule and PCM approach, the experimental data are reproduced within a few kcal mol(-)(1). Although the energy difference of the singlet and triplet spin states of the arylselanylium ions is small for the gas-phase structures, the singlet cation is undoubtedly the dominating species in solution because the triplet cation lacks the ability to form covalent bonds. PMID- 12590544 TI - Norbornyl cations of group 14 elements. AB - Norbornyl cations of the group 14 elements Si --> Pb have been synthesized from substituted 3-cyclopentenemethyl precursors by intramolecular addition of transient cations to the C=C double bond of the 3-cyclopentenemethyl substituent (pi-route to norbornyl cations). The norbornyl cations 4a (E = Si, R = Me), 4e (E = Si, R = Et), 4f (E = Si, R = Bu), 4g (E = Ge, R = Bu), 4h (E = Sn, R = Bu), and 4i (E = Pb, R = Et) have been identified by their characteristic NMR chemical shifts (4a,e,f, delta((29)Si) = 80-87, delta((13)C)(CH=) = 149.6-150.6; 4g, delta((13)C)(CH=) = 144.8; 4h, delta((119)Sn) = 334, delta((13)C)(CH=) = 141.5; 4i, delta((207)Pb) = 1049, delta((13)C)(CH=) = 138). The significant deshielding of the vinylic carbon atoms (Deltadelta((13)C)) relative to those of the precursor (Deltadelta((13)C) = 19.3-20.3 (4a,e,f), Deltadelta((13)C) = 14.6 (4g), Deltadelta((13)C) = 11.1 (4h), Deltadelta((13)C) approximately 8 (4i)) and the small J coupling constants between the element and the remote vinyl carbons in the case of 4h and 4i (J(CSn) = 26 Hz, J(CPb) = 16 Hz) give experimental evidence for the intramolecular interaction and the charge transfer between the positively charged element and the remote C=C double bond. The experimental results are supported by quantum mechanical calculations of structures, energies, and magnetic properties for the norbornyl cations 4a,b (E = Ge, R = Me), 4c (E = Sn, R = Me), 4d (E = Pb, R = Me), and 4e,f at the GIAO/B3LYP/6-311G(3d,p)//MP2/6 311G(d,p) (Si, Ge, C, H), SDD (Sn, Pb) level of theory. The calculated (29)Si NMR chemical shifts for the silanorbornyl cations 4a,e,f (delta((29)Si) = 77-93) agree well with experiment, and the calculated structures of the cations 4a-f reveal their bridged norbornyl cation nature and suggest also for the experimentally observed species 4a,e-i a formally 3 + 1 coordination for the element atom with the extra coordination provided by the C=C double bond. This places five carbon atoms in the close vicinity of the positively charged element atom. The group 14 element norbornyl cations 4a,e-i exhibit only negligible interactions with the aromatic solvent, and they are, depending on the nature of the element group, stable at room temperature in aromatic solvents for periods ranging from a few hours to days. In acetonitrile solution, the intramolecular interaction in the norbornyl cations 4a,e-h breaks down and nitrilium ions with the element in a tetrahedral environment are formed. In contrast, reaction of acetonitrile with the plumbyl cation 4i forms an acetonitrile complex, 10i, in which the norbornyl cation structure is preserved. The X-ray structure of 10i reveals a trigonal bipyramidal environment for the lead atom with the C=C double bond of the cyclopentenemethyl ligand and the nitrogen atom of the acetonitrile molecule in apical positions. Density functional calculations at the B3LYP/6 311G(2d,p)//(B3LYP/6-31G(d) (C, H), SDD (Si, Ge, Sn, Pb)) + DeltaZPVE level indicate that the thermodynamic stability of the group 14 norbornyl cations increases from Si to Pb. This results in a relative stabilization for the plumbanorbornyl cation 4d compared to tert-butyl cation of 52.7 kcal mol(-)(1). In contrast, the intramolecular stabilization energy E(A) of the norbornyl cations 4a-d decreases, suggesting reduced interaction between the C=C double bond and the electron-deficient element center in the plumbacation compared to the silacations. This points to a reduced electrophilicity of the plumbacation compared to its predecessors. PMID- 12590545 TI - Direct catalytic asymmetric aldol reaction of hydroxyketones: asymmetric Zn catalysis with a Et(2)Zn/linked-BINOL complex. AB - Full details of our newly developed catalyses with asymmetric zinc complexes as mimics of class II zinc-containing aldolase are described. A Et(2)Zn/(S,S)-linked BINOL complex was developed and successfully applied to direct catalytic asymmetric aldol reactions of hydroxyketones. A Et(2)Zn/(S,S)-linked-BINOL 1 = 2/1 system was initially developed, which efficiently promoted the direct aldol reaction of 2-hydroxy-2'-methoxyacetophenone (7d). Using 1 mol % of (S,S)-linked BINOL 1 and 2 mol % of Et(2)Zn, we obtained 1,2-dihydroxyketones syn-selectively in high yield (up to 95%), good diastereomeric ratio (up to 97/3), and excellent enantiomeric excess (up to 99%). Mechanistic investigation of Et(2)Zn/(S,S) linked-BINOL 1, including X-ray analysis, NMR analysis, cold spray ionization mass spectrometry (CSI-MS) analysis, and kinetic studies, provided new insight into the active oligomeric Zn/(S,S)-linked-BINOL 1/ketone 7d active species. On the basis of mechanistic investigations, a modified second generation Et(2)Zn/(S,S)-linked-BINOL 1 = 4/1 with molecular sieves 3A (MS 3A) system was developed as a much more effective catalyst system for the direct aldol reaction. As little as 0.1 mol % of (S,S)-linked-BINOL 1 and 0.4 mol % of Et(2)Zn promoted the direct aldol reaction smoothly, using only 1.1 equiv of 7d as a donor (substrate/ligand = 1000). This is the most efficient, in terms of catalyst loading, asymmetric catalyst for the direct catalytic asymmetric aldol reaction. Moreover, the Et(2)Zn/(S,S)-linked-BINOL 1 = 4/1 system was effective in the direct catalytic asymmetric aldol reaction of 2-hydroxy-2'-methoxypropiophenone (12), which afforded a chiral tetrasubstituted carbon center (tert-alcohol) in good yield (up to 97%) and ee (up to 97%), albeit in modest syn-selectivity. Newly developed (S,S)-sulfur-linked-BINOL 2 was also effective in the direct aldol reaction of 12. The Et(2)Zn/(S,S)-sulfur-linked-BINOL 2 = 4/1 system gave aldol adducts anti-selectively in good ee (up to 93%). Transformations of the aldol adducts into synthetically versatile intermediates were also described. PMID- 12590546 TI - Group 4 octahedral benzamidinate complexes: syntheses, structures, and catalytic activities in the polymerization of propylene modulated by pressure. AB - The synthesis and structural X-ray diffraction studies for some benzamidinate ligations and several group 4 benzamidinate complexes are presented. The use of the cis-octahedral C(2)-symmetry compounds was studied to shed light on the conceptual applicability of these complexes as potential catalysts for the stereoregular polymerization of propylene. We demonstrate that the stereoregular polymerization of propylene catalyzed by early-transition metal octahedral benzamidinate complexes, activated with either MAO or B(C(6)F(5))(3) as cocatalysts, can be modulated by pressure (from atactic to isotactic through elastomers). The different effects in the polymerization process such as the nature of solvent or cocatalyst, temperature, pressure, molar ratio catalyst:cocatalyst, and the relationship between the symmetry of the complex and the polymer microstructure have been investigated. When the complex [4-CH(3) C(6)H(4)C(NTMS)(2)](2)ZrMe(2) (9) was activated with MAO, it was found to be a good catalyst for the polymerization of propylene, at atmospheric pressure, producing an oily polymer resembling an atactic polypropylene. Being activated with B(C(6)F(5))(3), complex 9 produces a highly isotactic (mmmm = 98%) product. Likewise, when the polymerization of propylene was performed with complex 9 and MAO at high pressure (liquid propylene), a highly stereoregular polymer was also obtained. Larger activities and stereoregularities were achieved by performing the reaction in CH(2)Cl(2) as compared to toluene. Contrary to complex 9, at atmospheric pressure the complex [4-CH(3)-C(6)H(4)C(NTMS)(2)](2)TiMe(2) (10) is not active either in CH(2)Cl(2) or in toluene. At high pressure, complex 10 produces elastomeric polypropylene. Activities of the isolobal complexes [C(6)H(4)C(NTMS)(2)](2)ZrMe(2) (11) and [C(6)H(4)C(NTMS)(2)](2)TiMe(2) (12) were found to be larger than those of complexes 9 and 10, respectively. Contrary to the structures of the elastomeric polypropylenes described in the literature, the obtained elastomers are characterized by frequent alternation of the isotactic domains with stereodefects. The stereoregular errors are formed by the intramolecular epimerization of the growing chain at the last inserted unit. The epimerization reaction was corroborated through the isomerization of alkenes. PMID- 12590547 TI - Zeolite-confined Nano-RuO(2): A green, selective, and efficient catalyst for aerobic alcohol oxidation. AB - The development of green, selective, and efficient catalysts, which can aerobically oxidize a variety of alcohols to their corresponding aldehydes and ketones, is of both economic and environmental significance. We report here the synthesis of a novel aerobic oxidation catalyst, a zeolite-confined nanometer sized RuO(2) (RuO(2)-FAU), by a one-step hydrothermal method. Using the spatial constraints of the rigid zeolitic framework, we sucessfully incorporated RuO(2) nanoparticles (1.3 +/- 0.2 nm) into the supercages of faujasite zeolite. Ru K edge X-ray absorption fine structure results indicate that the RuO(2) nanoclusters anchored in the zeolite are structurally similar to highly hydrous RuO(2); that is, there is a two-dimensional structure of independent chains, in which RuO(6) octahedra are connected together by two shared oxygen atoms. In our preliminary catalytic studies, we find that the RuO(2) nanoclusters exhibit extraordinarily high activity and selectivity in the aerobic oxidation of alcohols under mild conditions, for example, air and ambient pressure. The physically trapped RuO(2) nanoclusters cannot diffuse out of the relatively narrow channels/pores of the zeolite during the catalytic process, making the catalyst both stable and reusable. PMID- 12590548 TI - Mechanically linked polycarbonate. AB - The synthesis, by solid-state copolymerization, and characterization of the first polycatenanes based on a commercial polymer are reported. Various amounts of a benzylic amide [2]catenane, the corresponding macrocycle, and a rigid bisphenol fluorene derivative have been quantitatively and homogeneously incorporated into bisphenol A polycarbonate. The resulting copolymers were characterized by size exclusion chromatography coupled with viscosimetry, (1)H NMR, differential scanning calorimetry, and dynamic mechanical analysis. The unexpectedly small influence of [2]catenane incorporation on the glass transition temperature of the copolymers points to remarkable internal mobility of the catenane comonomer rings. A new relaxation linked to the flexible catenane units is also observed. The studies represent a detailed structural characterization of a polymer containing small amounts of mechanical linkages in its backbone and demonstrate that significant effects can be induced by doping conventional polymers with small percentages (2-6% of repeat units) of flexible catenanes. PMID- 12590549 TI - Understanding the correlation of structure and selectivity in the chiral phosphoramide-catalyzed enantioselective allylation reactions: solution and solid state structural studies of bisphosphoramide.SnCl(4) complexes. AB - Complexation of bisphosphoramides, linked by various length methylene tethers, with tin tetrachloride was studied by both solution NMR spectroscopy and single crystal X-ray crystallography. The formation of cis-configured, octahedral 1/1 bisphosphoramide.SnCl(4) complexes was supported by both crystallographic and solution NMR studies. In addition, the formation of such complexes was shown to be highly dependent on the tether length and solution concentration. Single crystal X-ray analysis of bisphosphoramide.SnCl(4) complexes also provided detailed information on the stereochemical environment relevant to the enantioselectivity of asymmetric allylation reactions. PMID- 12590551 TI - The bis-barium complex of a butterfly crown ether as a phototunable supramolecular catalyst. AB - Reversible phototuning of the catalytic efficiency of the bis-barium complex of azobis(benzo-18-crown-6) in the basic ethanolysis of anilide derivatives has been achieved by light-induced cis right arrow over left arrow trans interconversion of the azobenzene spacer unit of the catalyst. The geometry of the productive catalyst-substrate complex is more favorable when the concave cis form of the catalyst is involved. Continuous photoregulation of the catalytic activity at any intermediate value between the "HIGH" and "LOW" levels was achieved by proper adjustment of the excitation wavelength or the irradiation time. The complete and relatively fast interconvertibility of photostationary states allowed the activity of the catalyst to be repeatedly photoswitched "HIGH" and "LOW" in the course of the same run. PMID- 12590550 TI - DNA-dye conjugates for controllable H aggregation(1). AB - Methyl Red H aggregate of predetermined size is successfully synthesized from the DNA conjugate involving multiple Methyl Red moieties in sequence. In the single stranded state, hypsochromicity monotonically increases with the number of incorporated dyes: the peak maximum of the conjugate involving six Methyl Reds appears at 415 nm, and the shift is as great as 69 nm (3435 cm(-)(1)) with respect to the monomeric transition. This large hypsochromicity accompanied by the narrowing of the band clearly demonstrates that H aggregate is formed in the single strand. H aggregation is further promoted at higher ionic strength. Upon addition of complementary DNA below the T(m), however, this H band disappears and a new peak appears at 448 nm, indicating that aggregated structure is changed by the duplex formation. This spectral change is completely reversible so that the H band at 415 nm appears again above T(m). Thus, aggregated structure can be reversibly controlled by the formation and dissociation of the DNA duplex. PMID- 12590552 TI - Modeling random methyl branching in ethylene/ propylene copolymers using metathesis chemistry: synthesis and thermal behavior. AB - The structure of random ethylene/propylene (EP) copolymers has been modeled using step polymerization chemistry. Six ethylene/propylene model copolymers have been prepared via acyclic diene metathesis (ADMET) polymerization and characterized for primary and higher level structure using in-depth NMR, IR, DSC, WAXD, and GPC analysis. These copolymers possess 1.5, 7.1, 13.6, 25.0, 43.3, and 55.6 methyl branches per 1000 carbons. Examination of these macromolecules by IR and WAXD analysis has demonstrated the first hexagonal phase in EP copolymers containing high ethylene content (90%) without the influence of sample manipulation (temperature, pressure, or radiation). Thermal behavior studies have shown that the melting point and heat of fusion decrease as the branch content increases. Further, comparisons have been made between these random ADMET EP copolymers, random EP copolymers made by typical chain addition techniques, and precisely branched ADMET EP copolymers. PMID- 12590553 TI - Cyclopentadienyl substituent effects on reductive elimination reactions in group 4 metallocenes: kinetics, mechanism, and application to dinitrogen activation. AB - The rate of reductive elimination for a family of zirconocene isobutyl hydride complexes, Cp(CpR(n)())Zr(CH(2)CHMe(2))H (Cp = eta(5)-C(5)Me(5), CpR(n)() = substituted cyclopentadienyl), has been measured as a function of cyclopentadienyl substituent. In general, the rate of reductive elimination increases modestly with the incorporation of sterically demanding substituents such as [CMe(3)] or [SiMe(3)]. A series of isotopic labeling experiments was used to elucidate the mechanism and rate-determining step for the reductive elimination process. From these studies, a new zirconocene isobutyl hydride complex, Cp' '(2)Zr(CH(2)CHMe(2))(H) (Cp' ' = eta(5)-C(5)H(3)-1,3-(SiMe(3))(2)), was designed and synthesized such that facile reductive elimination of isobutane and activation of dinitrogen was observed. The resulting dinitrogen complex, [Cp' '(2)Zr](2)(mu(2), eta(2),eta(2)-N(2)), has been characterized by X-ray diffraction and displays a bond length of 1.47 A for the N(2) ligand, the longest observed in any metallocene dinitrogen complex. Solution magnetic susceptibility demonstrates that [Cp' '(2)Zr](2)(mu(2), eta(2), eta(2)-N(2)) is a ground-state triplet, consistent with two Zr(III), d(1) centers. Mechanistic studies reveal that the dinitrogen complex is derived from the reaction of N(2) with the resulting cyclometalated zirconocene hydride rather than directly from reductive elimination of alkane. PMID- 12590554 TI - Induced alignment of a solution-cast discotic hexabenzocoronene derivative for electronic devices investigated by surface X-ray diffraction. AB - A surface X-ray diffraction study is presented showing that highly ordered and uniaxially aligned hexa(3,7-dimethyl-octanyl)hexa-peri-hexabenzocoronene (HBC C8,2) films can be fabricated by crystallization from solution onto friction transferred poly(tetrafluoroethylene) (PTFE) layers. Three crystalline HBC-C8,2 majority phases result. In all three phases, the HBC-C8,2 molecules self-organize into columns which are uniaxially aligned along the direction defined by the PTFE macromolecules of the substrate. The three phases are quite similar, the major difference being their orientation with respect to the substrate. A quasi-2D epitaxial growth mechanism with a grapho-epitaxial component for one of the three phases explains the formation of the three rotational HBC-C8,2 variants. A method to obtain a thin film with only one phase is proposed. The results show that standard THETAV;-2THETAV; X-ray diffraction and transmission electron diffraction can be very misleading tools to estimate the crystalline quality in a thin film of complex structure. PMID- 12590555 TI - Barium azacryptand sodide, the first alkalide with an alkaline Earth cation, also contains a novel dimer, (Na(2))(2-). AB - The first barium sodide, with stoichiometry Ba(2+)(H(5)Azacryptand[2.2.2](-))Na( ).2MeNH(2), was synthesized by the reaction of Ba, Na, and H(6)Azacryptand[2.2.2] in NH(3)-MeNH(2) solution. It was characterized by X-ray crystallography, (23)Na MAS NMR, hydrogen evolution, DSC, optical spectroscopy, and magnetic susceptibility. This is the first sodide in which the sodium anions form (Na(2))(2)(-) dimers. Previous theoretical predictions were verified by a calculation of the potential energy curve for the dimer in the field of the surrounding charges, whose positions were determined from the crystal structure. PMID- 12590556 TI - Enhancement of the ETS-10 titanosilicate activity in the shape-selective photocatalytic degradation of large aromatic molecules by controlled defect production. AB - In recent times, it has been shown that the microporous ETS-10 titanosilicate can be used as a shape-selective photocatalyst for the decomposition of aromatic molecules (Chem. Commun. 2001, 2131). Its actual use on practical grounds is however discouraged by its too low activity, when compared with that of TiO(2) photocatalysts. In the present work, we show how an ad hoc mild treatment with HF enhances the activity of ETS-10 toward the photodegradation of large aromatic molecules that are unable to penetrate inside the zeolitic pores, such as 2,5 dichlorophenol, 2,4,5-trichlorophenol, 1,3,5-trihydroxybenzene, and 2,3 dihydroxynaphthalene (DHN). The photoactivity of the acid-treated materials is comparable or even greater than that of the nonselective TiO(2) catalyst. Moreover, the enhancement of the photoactivity is accompanied by a remarkable parallel increase of the shape selectivity, particularly toward DHN (k(DHN)/k(P) = 127, where P = phenol). A complete characterization (by means of X-ray diffraction, scanning electron microscopy, transmission electron microscopy, ultraviolet-visible spectroscopy, and X-ray aborption spectroscopy techniques) of a set of ETS-10 samples which have undergone a progressively severe HF treatment allows us to propose an explanation of the photocatalytic activity and selectivity of the modified materials. PMID- 12590557 TI - Reactivity of substituted charged phenyl radicals toward components of nucleic acids. AB - Reactions of differently substituted phenyl radicals with components of nucleic acids have been investigated in the gas phase. A positively charged group located meta with respect to the radical site was employed to allow manipulation of the radicals in a Fourier-transform ion cyclotron resonance mass spectrometer. All of these electrophilic radicals react with sugars via exclusive hydrogen atom abstraction, with adenine and uracil almost exclusively via addition (likely at the C8 and C5 carbons, respectively), and with the nucleoside thymidine by hydrogen atom abstraction and addition at C5 in the base moiety (followed by elimination of (*)CH(3)). These findings parallel the reactivity of the phenyl radical with components of nucleic acids in solution, except that the selectivity for addition is different. Like HO(*), the electrophilic charged phenyl radicals appear to favor addition to the C5-end of the C5-C6 double bond of thymine and thymidine, whereas the phenyl radical preferentially adds to C6. The charged phenyl radicals do not predominantly add to thymine, as the neutral phenyl radical and HO(*), but mainly react by hydrogen atom abstraction from the methyl group (some addition to C5 in the base followed by loss of (*)CH(3) also occurs). Adenine appears to be the preferred target among the nucleobases, while uracil is the least favored. A systematic increase in the electrophilicity of the radicals by modification of the radicals' structures was found to facilitate all reactions, but the addition even more than hydrogen atom abstraction. Therefore, the least reactive radicals are most selective toward hydrogen atom abstraction, while the most reactive radicals also efficiently add to the base. Traditional enthalpy arguments do not rationalize the rate variations. Instead, the rates reflect the radicals' electron affinities used as a measure for their ability to polarize the transition state of each reaction. PMID- 12590558 TI - Electrochemical infrared characterization of CO comains on ruthenium-decorated platinum nanoparticles. AB - Spectra obtained by electrochemical infrared reflection absorption spectroscopy (EC-IRAS) for carbon monoxide (CO) adlayers formed by partial CO dosing on various ruthenium-decorated platinum nanoparticle films are reported. The need to achieve a well distributed rather than aggregated metal nanoparticle array is demonstrated, given that such nanoparticle aggregates induce complex dielectric behavior. The strategy here is to use an "organic glue matrix" (short chain SAMs) between the nanoparticles and the gold substrates. The observed promotion in CO electrooxidation by the existence of a Ru island on Pt nanoparticles, of interest to fuel-cell catalysis, showed a strong relationship with Ru surface concentrations, consistent with previous studies on single crystal or polycrystalline bimetallic surfaces. Two distinctive CO infrared bands, one for the Pt-CO and one for Ru-CO domain were found after the dipole coupling of CO within the two CO domains was minimized. Interestingly, those two CO bands showed independent electrooxidation behavior with electrode potential changes. Also, it is shown that the electrooxidation of CO on large Ru islands is less facile than on small Ru islands. In addition, the activity of commercial Pt/Ru alloy nanoparticles to CO stripping was tested and IRAS spectra were reported as a comparison to our Ru-decorated Pt nanoparticles. PMID- 12590559 TI - Is the ruthenium analogue of compound I of cytochrome p450 an efficient oxidant? A theoretical investigation of the methane hydroxylation reaction. AB - High-valent metal-oxo complexes catalyze C-H bond activation by oxygen insertion, with an efficiency that depends on the identity of the transition metal and its oxidation state. Our study uses density functional calculations and theoretical analysis to derive fundamental factors of catalytic activity, by comparison of a ruthenium-oxo catalyst with its iron-oxo analogue toward methane hydroxylation. The study focuses on the ruthenium analogue of the active species of the enzyme cytochrome P450, which is known to be among the most potent catalysts for C-H activation. The computed reaction pathways reveal one high-spin (HS) and two low spin (LS) mechanisms, all nascent from the low-lying states of the ruthenium-oxo catalyst (Ogliaro, F.; de Visser, S. P.; Groves, J. T.; Shaik, S. Angew. Chem. Int. Ed. 2001, 40, 2874-2878). These mechanisms involve a bond activation phase, in which the transition states (TS's) appear as hydrogen abstraction species, followed by a C-O bond making phase, through a rebound of the methyl radical on the metal-hydroxo complex. However, while the HS mechanism has a significant rebound barrier, and hence a long lifetime of the radical intermediate, by contrast, the LS ones are effectively concerted with small barriers to rebound, if at all. Unlike the iron catalyst, the hydroxylation reaction for the ruthenium analogue is expected to follow largely a single-state reactivity on the LS surface, due to a very large rebound barrier of the HS process and to the more efficient spin crossover expected for ruthenium. As such, ruthenium-oxo catalysts (Groves, J. T.; Shalyaev, K.; Lee, J. In The Porphyrin Handbook; Biochemistry and Binding: Activation of Small Molecules, Vol. 4; Kadish, K. M., Smith, K. M., Guilard, R., Eds.; Academic Press: New York, 2000; pp 17-40) are expected to lead to more stereoselective hydroxylations compared with the corresponding iron-oxo reactions. It is reasoned that the ruthenium-oxo catalyst should have larger turnover numbers compared with the iron-oxo analogue, due to lesser production of suicidal side products that destroy the catalyst (Ortiz de Montellano, P. R.; Beilan, H. S.; Kunze, K. L.; Mico, B. A. J. Biol. Chem. 1981, 256, 4395-4399). The computations reveal also that the ruthenium complex is more electrophilic than its iron analogue, having lower hydrogen abstraction barriers. These reactivity features of the ruthenium-oxo system are analyzed and shown to originate from a key fundamental factor, namely, the strong 4d(Ru)-2p(O,N) overlaps, which produce high-lying pi(Ru-O), sigma(Ru-O), and sigma(Ru-N) orbitals and thereby to lead to a preference of ruthenium for higher-valent oxidation states with higher electrophilicity, for the effectively concerted LS hydroxylation mechanism, and for less suicidal complexes. As such, the ruthenium oxo species is predicted to be a more robust catalyst than its iron-oxo analogue. PMID- 12590560 TI - Bonding of atomic phosphorus to polycyclic hydrocarbons and curved graphitic surfaces. AB - We present a theoretical study of the bonding of atomic phosphorus to planar hydrocarbons and to curved graphite-like surfaces. We find that bonding of phosphorus to planar polycyclic hydrocarbons induces curvature away from the phosphorus atom, as defined by the pyramidalization angle. Similarly, bonding of atomic phosphorus to the [5,5] fulvalene-circulene semifullerene and buckminsterfullerene is only possible on the convex side of the carbon surface. On the other hand, we find the interaction of atomic phosphorus with the concave side of fullerene-like surfaces to be nonbonding for both quartet and doublet spin states. We find the prerequisite for stable epoxy-type bonds within these systems is the ability of the carbon atoms to maintain or induce curvature away from the P.C=C bond. PMID- 12590561 TI - Mechanism of a chemical classic: quantum chemical investigation of the autocatalyzed reaction of the serendipitous wohler synthesis of urea. AB - The detailed reaction pathways for the ammonium cyanate transformation into urea (Wohler's reaction) in the gas phase, in solution, and in the solid state have exhaustively been explored by means of first-principles quantum chemical calculations at the B3LYP level of theory using the 6-31G(d,p) basis set. This serendipitous synthesis of urea is predicted to proceed in two steps; the first step involves the decomposition of the ammonium cyanate to ammonia and isocyanic or cyanic acid, and the second one, which is the main reaction step (and probably the rate-determining step), involves the interaction of NH(3) with either isocyanic or cyanic acid. Several alternative pathways were envisaged for the main reaction step of Wohler's reaction in a vacuum involving the formation of "four-center" transition states. Modeling Wohler's reaction in aqueous solution and in the solid state, it was found that the addition of NH(3) to both acids is assisted (autocatalyzed) by the active participation of extra H(2)O and/or NH(3) molecules, through a preassociative, cooperative, and hydrogen-transfer relay mechanism involving the formation of "six-center" or even "eight-center" transition states. The most energetically economic path of the rate-determining step of Wohler's reaction is that of the addition of NH(3) to the C=N double bond of isocyanic acid, directly affording urea. An alternative pathway corresponding to the anti-addition of ammonia to the Ctbd1;N triple bond of cyanic acid, yielding urea's tautomer HN=C(OH)NH(2), seems to be another possibility. In the last case, urea is formed through a prototropic tautomerization of its enolic form. The energies of the reactants, products, and all intermediates along with the barrier heights for each reaction path have been calculated at the B3LYP/6 31G(d,p) level of theory. The geometry optimization and characterization of all of the stationary points found on the potential energy hypersurfaces was performed at the same level of theory. PMID- 12590562 TI - Mechanism of charging and supercharging molecules in electrospray ionization. AB - The origin of the extent of charging and the mechanism by which multiply charged ions are formed in electrospray ionization have been hotly debated for over a decade. Many factors can affect the number of charges on an analyte ion. Here, we investigate the extent of charging of poly(propyleneimine) dendrimers (generations 3.0 and 5.0), cytochrome c, poly(ethylene glycol)s, and 1,n diaminoalkanes formed from solutions of different composition. We demonstrate that in the absence of other factors, the surface tension of the electrospray droplet late in the desolvation process is a significant factor in determining the overall analyte charge. For poly(ethylene glycol)s, 1,n-diaminoalkanes, and poly(propyleneimine) dendrimers electrosprayed from single-component solutions, there is a clear relationship between the analyte charge and the solvent surface tension. Addition of m-nitrobenzyl alcohol (m-NBA) into electrospray solutions increases the charging when the original solution has a lower surface tension than m-NBA, but the degree of charging decreases when this compound is added to water, which has a higher surface tension. Similarly, the charging of cytochrome c ions formed from acidified denaturing solutions generally increases with increasing surface tension of the least volatile solvent. For the dendrimers investigated, there is a strong correlation between the average charge state of the dendrimer and the Rayleigh limiting charge calculated for a droplet of the same size as the analyte molecule and with the surface tension of the electrospray solvent. A bimodal charge distribution is observed for larger dendrimers formed from water/m-NBA solutions, suggesting the presence of more than one conformation in solution. A similar correlation is found between the extent of charging for 1,n-diaminoalkanes and the calculated Rayleigh limiting charge. These results provide strong evidence that multiply charged organic ions are formed by the charged residue mechanism. A significantly smaller extent of charging for both dendrimers and 1,n-diaminoalkanes would be expected if the ion evaporation mechanism played a significant role. PMID- 12590563 TI - Structural and vibrational study of the tautomerism of histamine free-base in solution. AB - Infrared and Raman spectroscopy in H(2)O and D(2)O and quantum Density Functional calculations were used to determine the structure of histamine free-base in aqueous solution. A quantum mechanical study of the tautomeric equilibrium of histamine free-base in solution was performed at the 6-311G level. Electronic correlation energies were included by using the hybrid functional B3LYP. The solvent was simulated as a continuum characterized by a dielectric constant, and the quantum system (solute) was placed in an ellipsoidal cavity. Solute-solvent electrostatic interaction was calculated by means a multipolar moment expansion introduced in the Hamiltonian. Four relevant histamine conformations were optimized by allowing all the geometrical parameters to vary independently, which involved both the gauche-trans and the N3H-N1H tautomerisms. The calculated free energies predicted N3H-gauche as the most stable one of histamine free-base in solution. The order of stability is here completely altered with respect to previous results in gas phase, which presented the N1H-gauche conformer as the most stable structure. Our results also differ from previous Monte Carlo simulations, which obtained the N3H-trans conformer as the most stable in solution, although in this case, the histamine structures were kept frozen to the gas-phase geometry. Vibrational spectroscopy results support theoretical ones. Quadratic force fields for the four histamine conformers were achieved under the same calculation methodology. Previously, a general assignment of the infrared and Raman spectra of histamine free-base was proposed for solutions in both natural and heavy water. This allowed us to compare the experimental set of both wavenumbers and infrared intensities with the calculated ones. The lowest quadratic mean wavenumber deviation was obtained for the N3H-gauche conformer, in agreement with the free-energy calculations. Calculated infrared intensities were also compared to the experimental intensities, supporting this conformer as the relevant structure of histamine free-base in solution. It was then selected for a complete vibrational dynamics calculation, starting with a low-level scaling procedure to fit the set of calculated wavenumbers to the experimental values. The results were presented in terms of quadratic force constants, potential energy distribution, and normal modes. PMID- 12590564 TI - Reaction pathways of the Simmons-Smith reaction. AB - The cyclopropanation reaction of an alkene with a metal carbenoid has been studied by means of the B3LYP hybrid density functional method. The cyclopropanation of ethylene with a lithium carbenoid or a zinc carbenoid [Simmons-Smith (SS) reagent] goes through two competing pathways, methylene transfer and carbometalation. Both processes are fast for the lithium carbenoid, while, for the zinc carbenoid, only the former is fast enough to be experimentally feasible. The reaction of an SS reagent (ClZnCH(2)Cl) with ethylene and an allyl alcohol in the presence of ZnCl(2) was also studied. The allyl alcohol reaction was modeled with an SS reagent/alkoxide complex (ClCH(2)ZnOCH(2)CH=CH(2)) formed from the SS reagent and allyl alcohol. Two modes of acceleration were found. The first involves the well-accepted mechanism of 1,2 chlorine migration, and the second involves a five-centered bond alternation. The latter was found to be more facile than the former and to operate equally well both with ethylene and with aggregates of SS reagent/alkoxide complexes. Calculations on the SS reaction with 2-cyclohexen-1-ol offer a reasonable model for the hydroxy-directed diastereoselective SS reaction, which has been used for a long time in organic synthesis. PMID- 12590565 TI - Impact of multiple cation-pi interactions upon calix[4]arene substrate binding and specificity. AB - The cation-pi interaction influence on the conformation and binding of calix[4]arenes to alkali-metal cations has been studied using a dehydroxylated model. The model allows for the separation of cooperative cation-pi and electrostatic forces commonly found in the binding motifs found in calixarene complexes. Starting from the four well-known calix[4]arene conformations, six conformers for this dehydroxylated model (cone, partial cone, flattened cone, chair, 1,2-alternate, and 1,3-alternate) have been characterized by geometry optimization and frequency analysis using the Becke three-parameter exchange functional with the nonlocal correlation functional of Lee, Yang, and Parr and the 6-31G(d) basis set. Without the stabilization provided by the hydroxyl hydrogen bonds in calix[4]arene, neither the cone nor the 1,2-alternate conformation is computed to be a ground-state structure. The partial cone, flattened cone, chair, and 1,3-alternate conformers have been identified as ground-state structures in a vacuum, with the partial cone and the 1,3-alternate as the lowest energy minima in the aromatic model. The C(4)(v)() cone conformation is found to be a transition structure separating the flattened cone (C(2)(v)()) conformers. The energetic and structural preferences of the calix[4]arene model change dramatically when it is bound to Li(+), Na(+), and K(+). The number of pi-faces, the positioning of these pi-faces with respect to the cations, and the nature of the cation were studied as factors in the binding strength. A detailed study of the distances and angles between the aromatic ring centroids and the cations reveals the energetic advantages of multiple weak cation-pi interactions. The geometries are often far from the optimal cation-pi interaction in which the cation approaches in a perpendicular path the aromatic ring center, where the quadrupole moment is strongest. The results reveal that multiple weaker nonoptimal cation-pi interactions contribute significantly to the overall binding strength. This theoretical analysis underscores the importance of neighboring aromatic faces and provides new insight into the significance of cation-pi binding, not only for calix[4]arenes, but also for other supramolecular and biological systems. PMID- 12590566 TI - Memory effects based on intermolecular photoinduced proton transfer. AB - We have identified a strategy to communicate a chemical signal between two independent molecular components. One of them is a photoactive merocyanine that switches to a spiropyran, releasing a proton, when stimulated with visible light. The other is a 4,4'-pyridylpyridinium monocation that captures the released proton, producing an electroactive 4,4'-bipyridinium dication. Under the irradiation conditions employed, the photoinduced transformation requires ca. 15 min to reach a photostationary state. In the dark, the ensemble of communicating molecules reequilibrates to the original state in ca. 5 days. These processes can be monitored following the photoinduced enhancement and thermal decay, respectively, of the current for the monolectronic reduction of the 4,4' bipyridinium dication. The pronounced difference in time scale for the current enhancement and decay steps can be exploited to implement a memory element with a bit retention time of 11 h. A bit of information can be written optically in the chemical system and it can be read electrically and nondestructively. The memory can be reset, extending its permanence in the dark beyond the bit retention time. A binary logic analysis of the signal transduction operated by the communicating molecules reveals the characteristic behavior of sequential logic operators, which are the basic components of digital memories. PMID- 12590567 TI - Implications for RNase L in prostate cancer biology. AB - Recently, the interferon (IFN) antiviral pathways and prostate cancer genetics and have surprisingly converged on a single-strand specific, regulated endoribonuclease. Genetics studies from several laboratories in the U.S., Finland, and Israel, support the recent identification of the RNase L gene, RNASEL, as a strong candidate for the long sought after hereditary prostate cancer 1 (HPC1) allele. Results from these studies suggest that mutations in RNASEL predispose men to an increased incidence of prostate cancer, which in some cases reflect more aggressive disease and/or decreased age of onset compared with non-RNASEL linked cases. RNase L is a uniquely regulated endoribonuclease that requires 5'-triphosphorylated, 2',5'-linked oligoadenylates (2-5A) for its activity. The presence of both germline mutations in RNASEL segregating with disease within HPC-affected families and loss of heterozygosity (LOH) in tumor tissues suggest a novel role for the regulated endoribonuclease in the pathogenesis of prostate cancer. The association of mutations in RNASEL with prostate cancer cases further suggests a relationship between innate immunity and tumor suppression. It is proposed here that RNase L functions in counteracting prostate cancer by virtue of its ability to degrade RNA, thus initiating a cellular stress response that leads to apoptosis. This monograph reviews the biochemistry and genetics of RNase L as it relates to the pathobiology of prostate cancer and considers implications for future screening and therapy of this disease. PMID- 12590568 TI - Oxygen and hydrogen isotope effects in an active site tyrosine to phenylalanine mutant of peptidylglycine alpha-hydroxylating monooxygenase: mechanistic implications. AB - Peptidylglycine alpha-hydroxylating monooxygenase (PHM) and dopamine beta monooxygenase (DbetaM) are homologous copper-containing enzymes that catalyze an oxygen-dependent hydroxylation of peptide-extended glycine residues and phenethylamines, respectively. The mechanism whereby these enzymes activate molecular oxygen and the C-H bond of substrate has been the subject of numerous studies, and various mechanisms have been put forth. From the magnitude of (18)O isotope effects as a function of substrate structure in DbetaM, an active site tyrosine had been proposed to function in the reductive activation of Cu(II)-OOH to generate a reactive copper-oxo species [Tian et al. (1994) Biochemistry 33, 226]. The presence of a tyrosine residue, Y318, in the active site of PHM was subsequently confirmed from crystallographic studies [Prigge et al. (1997) Science 278, 1300]. We now report extensive kinetic and isotope effect studies on the Y318F mutant form of PHM, analyzing the role of this tyrosine in the catalytic mechanism. It is found that the Y318F mutant has intrinsic hydrogen and (18)O isotope effects that are within experimental error of the wild-type enzyme and that the mutation causes only a slight reduction in the rate constant for C-H bond cleavage. These findings, together with the recent demonstration that C-H activation in PHM is dominated by quantum mechanical tunneling [Francisco et al. (2002) J. Am. Chem. Soc. 124, 8194], necessitate a reexamination of plausible mechanisms for this unique class of copper enzymes. PMID- 12590569 TI - Structural and kinetic analysis of catalysis by a thiamin diphosphate-dependent enzyme, benzoylformate decarboxylase. AB - Benzoylformate decarboxylase is a member of the family of enzymes that are dependent on the cofactor thiamin diphosphate. A structure of this enzyme binding (R)-mandelate, a competitive inhibitor, suggests that at least two hydrogen bonds are formed between the substrate, benzoylformate, and active site side chains. The first is between the carboxylate group of benzoylformate and the hydroxyl group of S26, and the second is between carbonyl group of the substrate and an imidazole nitrogen of H70. Steady-state kinetic studies indicate that the catalytic parameters are strongly affected in three active site mutants, S26A, H70A, and H281A. The K(m) of S26A was increased most dramatically, 25-fold more than that of the wild-type enzyme, while the K(i) of (R)-mandelate was increased 100-fold, suggesting that the serine hydroxyl is important for substrate binding. The k(cat) of H70A is reduced more than 3 orders of magnitude, strongly implicating this residue in catalysis, and H281 showed significant, but smaller magnitude, effects on both K(m) and k(cat). Stopped-flow experiments using an alternative substrate, p-nitrobenzoylformate, lead to kinetic resolution of the fate of key thiamin diphosphate-bound intermediates. Together, the experimental results suggest the following roles for residues in the active site. The residue H70 is important for the protonation of the 2-alpha-mandelyl-ThDP intermediate, thereby assisting in decarboxylation, and for the deprotonation of the 2-alpha hydroxybenzyl-ThDP intermediate, aiding product release. H281 is involved in protonation of the enamine. Surprisingly, S26 appears to be involved not only in substrate binding but also in other steps of the reaction. PMID- 12590570 TI - Characterization of a mutant Bacillus subtilis adenylosuccinate lyase equivalent to a mutant enzyme found in human adenylosuccinate lyase deficiency: asparagine 276 plays an important structural role. AB - Adenylosuccinate lyase, an enzyme catalyzing two reactions in purine biosynthesis (the cleavage of either adenylosuccinate or succinylaminoimidazole carboxamide ribotide), has been implicated in a human disease arising from point mutations in the gene encoding the enzyme. Asn(276) of Bacillus subtilis adenylosuccinate lyase, a residue corresponding to the location of a human enzyme mutation, was replaced by Cys, Ser, Ala, Arg, and Glu. The mutant enzymes exhibit decreased V(max) values (2-400-fold lower) for both substrates compared to the wild-type enzyme and some changes in the pH dependence of V(max) but no loss in affinity for adenylosuccinate. Circular dichroism reveals no difference in secondary structure between the wild-type and mutant enzymes. We show here for the first time that wild-type adenylosuccinate lyase exhibits a protein concentration dependence of molecular weight, secondary structure, and specific activity. An equilibrium constant between the dimer and tetramer was measured by light scattering for the wild-type and mutant enzymes. The equilibrium is somewhat shifted toward the tetramer in the mutant enzymes. The major difference between the wild-type and mutant enzymes appears to be in quaternary structure, with many mutant enzymes exhibiting marked thermal instability relative to the wild-type enzyme. We propose that mutations at position 276 result in structurally impaired adenylosuccinate lyases which are assembled into defective tetramers. PMID- 12590571 TI - A novel modeling protocol for protein receptors guided by bound-ligand conformation. AB - A novel protocol for protein homology modeling is described, in which uncertainty in protein structure is resolved by applying the criterion that the protein must bind to a complementary ligand. A diverse library of protein models is created and then screened by docking with ligands of known conformation. The more accurate protein models form higher-quality docked complexes, and the quality of the fit is used to select the best models. The effectiveness of this technique with both natural and unnatural ligands is demonstrated by modeling the Fv fragment of an antibody and comparing the results to known crystal structures. PMID- 12590572 TI - O2- and alpha-ketoglutarate-dependent tyrosyl radical formation in TauD, an alpha keto acid-dependent non-heme iron dioxygenase. AB - Taurine/alpha-ketoglutarate dioxygenase (TauD), a non-heme mononuclear Fe(II) oxygenase, liberates sulfite from taurine in a reaction that requires the oxidative decarboxylation of alpha-ketoglutarate (alphaKG). The lilac-colored alphaKG-Fe(II)TauD complex (lambda(max) = 530 nm; epsilon(530) = 140 M(-)(1) x cm(-)(1)) reacts with O(2) in the absence of added taurine to generate a transient yellow species (lambda(max) = 408 nm, minimum of 1,600 M(-)(1) x cm( )(1)), with apparent first-order rate constants for formation and decay of approximately 0.25 s(-)(1) and approximately 0.5 min(-)(1), that transforms to yield a greenish brown chromophore (lambda(max) = 550 nm, 700 M(-)(1) x cm( )(1)). The latter feature exhibits resonance Raman vibrations consistent with an Fe(III) catecholate species presumed to arise from enzymatic self-hydroxylation of a tyrosine residue. Significantly, (18)O labeling studies reveal that the added oxygen atom derives from solvent rather than from O(2). The transient yellow species, identified as a tyrosyl radical on the basis of EPR studies, is formed after alphaKG decomposition. Substitution of two active site tyrosine residues (Tyr73 and Tyr256) by site-directed mutagenesis identified Tyr73 as the likely site of formation of both the tyrosyl radical and the catechol-associated chromophore. The involvement of the tyrosyl radical in catalysis is excluded on the basis of the observed activity of the enzyme variants. We suggest that the Fe(IV) oxo species generally proposed (but not yet observed) as an intermediate for this family of enzymes reacts with Tyr73 when substrate is absent to generate Fe(III) hydroxide (capable of exchanging with solvent) and the tyrosyl radical, with the latter species participating in a multistep TauD self-hydroxylation reaction. PMID- 12590573 TI - Generation of an active monomer of rabbit muscle creatine kinase by site-directed mutagenesis: the effect of quaternary structure on catalysis and stability. AB - Cytosolic creatine kinase exists in native form as a dimer; however, the reasons for this quaternary structure are unclear, given that there is no evidence of active site communication and more primitive guanidino kinases are monomers. Three fully conserved residues found in one-half of the dimer interface of the rabbit muscle creatine kinase (rmCK) were selectively changed to alanine by site directed mutagenesis. Four mutants were prepared, overexpressed, and purified: R147A, R151A, D209A, and R147A/R151A. Both the R147A and R147A/R151A were confirmed by size-exclusion chromatography and analytical ultracentrifugation to be monomers, whereas R151A was dimeric and D209A appeared to be an equilibrium mixture of dimers and monomers. Kinetic analysis showed that the monomeric mutants, R147A and R147A/R151A, showed substantial enzymatic activity. Substrate binding affinity by R147A/R151A was reduced approximately 10-fold, although k(cat) was 60% of the wild-type enzyme. Unlike the R147A/R151A, the kinetic data for the R147A mutant could not be fit to a random-order rapid-equilibrium mechanism characteristic of the wild-type, but could only be fit to an ordered mechanism with creatine binding first. Substrate binding affinities were also significantly lower for the R147A mutant, but k(cat) was 11% that of the native enzyme. Fluorescence measurements using 1-anilinonaphthalene-8-sufonate showed that increased amounts of hydrophobic surface area are exposed in all of the mutants, with the monomeric mutants having the greatest amounts of unfolding. Thermal inactivation profiles demonstrated that protein stability is significantly decreased in the monomeric mutants compared to wild-type. Denaturation experiments measuring lambda(max) of the intrinsic fluorescence as a function of guanidine hydrochloride concentration helped confirm the quaternary structures and indicated that the general unfolding pathway of all the mutants are similar to that of the wild-type. Collectively, the data show that dimerization is not a prerequisite for activity, but there is loss of structure and stability upon formation of a CK monomer. PMID- 12590574 TI - Global structure and dynamics of human apolipoprotein CII in complex with micelles: evidence for increased mobility of the helix involved in the activation of lipoprotein lipase. AB - Apolipoprotein CII (apoCII), a surface constituent of plasma lipoproteins, is the activator for lipoprotein lipase (LPL) and is therefore central for lipid transport in blood. The three-dimensional structure of (13)C-, (15)N-enriched human full-length apoCII in complex with sodium dodecyl sulfate (SDS) micelles is reported. In addition to the structure determination, (15)N-relaxation measurements have been performed at two magnetic fields to characterize the dynamics of the backbone of apoCII in the complex. The relaxation data also provided global structural constraints, viz. the orientation of helices in the complex. In addition, global constraints were derived from the fact that apoCII helices are attached to the surface of the SDS micelle and that the hydrophobic moments of each helix faces the interior of the micelle. These three categories of global constraints, together with the local classical NMR constraints, were sufficient to define the 3D structure of the apoCII-SDS micelle complex. To our knowledge, this presents the first example in which the global structure of a protein-SDS micelle complex has been determined. The C-terminal helix of apoCII is known to be responsible for the activation of LPL. This helix is distinguished from the other helices by a higher degree of internal motion on the nanosecond time scale as shown by the relaxation data. The overall structure and the internal dynamics, combined with previous mutation data, give important clues toward a possible mechanism for the activation of LPL by apoCII. PMID- 12590575 TI - Dynamic properties of the G93A mutant of copper-zinc superoxide dismutase as detected by NMR spectroscopy: implications for the pathology of familial amyotrophic lateral sclerosis. AB - The backbone assignment of the copper-zinc superoxide dismutase amyotrophic lateral sclerosis G93A mutant was performed on an (15)N-enriched protein sample. (15)N R(1), R(2), and R(1)(rho) and (15)N-(1)H NOE experiments were then carried out at 600 MHz on G93A Cu(2)Zn(2)SOD and the values compared to the dynamics data for the "wild-type" protein. In addition, (15)N and (1)H chemical shift comparisons between wild-type Cu(2)Zn(2)SOD and its G93A mutant were also made. G93A exhibits a higher mobility than wild-type Cu(2)Zn(2)SOD, particularly in loops III and V, on a time scale faster than the rate of protein tumbling. There are also distinct chemical shift and NOE differences in residues 35-42 and 92-95, which comprise these loops. These two regions of Cu(2)Zn(2)SOD form the end of the beta-barrel termed the "beta-barrel plug" [Tainer, J. A., Getzoff, E. D., Beem, K. M., Richardson, J. S., and Richardson, D. C. (1982) J. Mol. Biol. 160, 181-217]. The increased mobility and reduction of the number of observed NOEs in this region indicate an opening of the beta-barrel that may lead to amyloid fibrillogenesis. Alternatively, a motor neuron-specific substrate may bind this region of the protein, leading to deleterious modifications and/or reactions. PMID- 12590576 TI - Catalytic strategy of S-adenosyl-L-homocysteine hydrolase: transition-state stabilization and the avoidance of abortive reactions. AB - S-Adenosylhomocysteine hydrolase (AdoHcy hydrolase) crystallizes from solutions containing the intermediate analogue neplanocin A with the analogue bound in its 3'-keto form at the active sites of all of its four subunits and the four tightly bound cofactors in their reduced (NADH) state. The enzyme is in the closed conformation, which corresponds to the structure in which the catalytic chemistry occurs. Examination of the structure in the light of available, very detailed kinetic studies [Porter, D. J., Boyd, F. L. (1991) J. Biol. Chem. 266, 21616 21625. Porter, D. J., Boyd, F. L. (1992) J. Biol. Chem. 267, 3205-3213. Porter, D. J. (1998) J. Biol. Chem. 268, 66-73] suggests elements of the catalytic strategy of AdoHcy hydrolase for acceleration of the reversible conversion of AdoHcy to adenosine (Ado) and homocysteine (Hcy). The enzyme, each subunit of which possesses a substrate-binding domain that in the absence of substrate is in rapid motion relative to the tetrameric core of the enzyme, first binds substrate and ceases motion. Probably concurrently with oxidation of the substrate to its 3'-keto form, the closed active site is "sealed off" from the environment, as indicated by a large (10(8)(-)(9)-fold) reduction in the rate of departure of ligands, a feature that prevents exposure of the labile 3'-keto intermediates to the aqueous environment. Elimination of the 5'-substituent (Hcy in the hydrolytic direction, water in the synthetic direction) generates the central intermediate 4',5'-didehydro-5'-deoxy-3'-ketoadenosine. Abortive 3'-reduction of the central intermediate is prevented by a temporary suspension of all or part of the redox catalytic power of the enzyme during the existence of the central intermediate. The abortive reduction is 10(4)-fold slower than the productive reductions at the ends of the catalytic cycle and has a rate constant similar to those of nonenzymic intramolecular model reactions. The mechanism for suspending the redox catalytic power appears to be a conformationally induced increase in the distance across which hydride transfer must occur between cofactor and substrate, the responsible conformational change again being that which "seals" the active site. The crystal structure reveals a well-defined chain of three water molecules leading from the active site to the subunit surface, which may serve as a relay for proton exchange between solvent and active site in the closed form of the enzyme, permitting maintenance of active-site functional groups in catalytically suitable protonation states. PMID- 12590577 TI - Conformational dynamics of DnaB helicase upon DNA and nucleotide binding: analysis by intrinsic tryptophan fluorescence quenching. AB - DnaB helicase of E. coli unwinds duplex DNA in the replication fork using the energy of ATP hydrolysis. We have analyzed structural and conformational changes in the DnaB protein in various nucleotides and DNA bound intermediate states by fluorescence quenching analysis of intrinsic fluorescence of native tryptophan (Trp) residues in DnaB. Fluorescence quenching analysis indicated that Trp48 in domain alpha is in a hydrophobic environment and resistant to fluorescence quenchers such as potassium iodide (KI). In domain beta, Trp294 was found to be in a partially hydrophobic environment, whereas Trp456 in domain gamma appeared to be in the least hydrophobic environment. Binding of oligonucleotides to DnaB helicase resulted in a significant attenuation of the fluorescence quenching profile, indicating a change in conformation. ATPgammaS or ATP binding appeared to lead to a conformation in which Trp residues had a higher degree of solvent exposure and fluorescence quenching. However, the most dramatic increase of Trp fluorescence quenching was observed with ADP binding with a possible conformational relaxation. Site-specific Trp --> Cys mutants of DnaB helicase demonstrated that conformational change upon ADP binding could be attributed exclusively to a conformational transition in the alpha domain leading to an increase in the solvent exposure of Trp48. However, formation of DnaB.ATPgammaS.DNA ternary complex led to a conformation with a fluorescence quenching profile similar to that observed with DnaB alone. The DnaB.ADP.DNA ternary complex produced a quenching curve similar to that of DnaB.ADP complex pointing to a change in conformation due to ATP hydrolysis. There are at least four identifiable structural/conformational states of DnaB helicase that are likely important in the helicase activity. The noncatalytic alpha domain in the N terminus appeared to undergo the most significant conformational changes during nucleotide binding and hydrolysis. This is the first reported elucidation of the putative role of domain alpha, which is essential for DNA helicase action. We have correlated these results with partial structural models of alpha, beta, and gamma domains PMID- 12590578 TI - Powering DNA repair through substrate electrostatic interactions. AB - The reaction catalyzed by the DNA repair enzyme uracil DNA glycosylase (UDG) proceeds through an unprecedented stepwise mechanism involving a positively charged oxacarbenium ion sugar and uracil anion leaving group. Here we use a novel approach to evaluate the catalytic contribution of electrostatic interactions between four essential phosphodiester groups of the DNA substrate and the cationic transition state. Our strategy was to substitute each of these phosphate groups with an uncharged (R)- or (S)-methylphosphonate linkage (MeP). We then compared the damaging effects of these methylphosphonate substitutions on catalysis with their damaging effects on binding of a cationic 1-azadeoxyribose (1-aza-dR(+)) oxacarbenium ion analogue to the UDG-uracil anion binary complex. A plot of log k(cat)/K(m) for the series of MeP-substituted substrates against log K(D) for binding of the 1-aza-dR(+) inhibitors gives a linear correlation of unit slope, confirming that the electronic features of the transition state resemble that of the 1-aza-dR(+), and that the anionic backbone of DNA is used in transition state stabilization. We estimate that all of the combined phosphodiester interactions with the substrate contribute 6-8 kcal/mol toward lowering the activation barrier, a stabilization that is significant compared to the 16 kcal/mol catalytic power of UDG. However, unlike groups of the enzyme that selectively stabilize the charged transition state by an estimated 7 kcal/mol, these phosphodiester groups also interact strongly in the ground state. To our knowledge, these results provide the first experimental evidence for electrostatic stabilization of a charged enzymatic transition state and intermediate using the anionic backbone of DNA. PMID- 12590579 TI - Solution structure and DNA binding property of the fifth HMG box domain in comparison with the first HMG box domain in human upstream binding factor. AB - Human upstream binding factor (hUBF) is a nucleolar transcription factor involved in transcription by RNA polymerase I. It contains six HMG box domains. The contribution of each HMG box motif to its function is different. hUBF HMG box 1 shows a very strong binding affinity for both the four-way DNA junction and a 15 bp GC-rich rRNA gene core promoter fragment, but hUBF HMG box 5 shows a much weaker binding affinity for the four-way DNA junction and the GC-rich rRNA gene core promoter fragment. To illustrate the molecular basis of their DNA binding difference, the solution structure of box 5 was studied by NMR. The tertiary structure of box 5 shows a common flattened L-shaped fold, similar to box 1 and other HMG boxes with known structures. It is formed by intersection of three helical arms: helix 1 (residues 9-25) and helix 2 (residues 30-42) pack into each other to form the major wing, while helix 3 (residues 48-70) is aligned with the extended N-terminal segment to form the minor wing. A hydrophobic core is formed by three tryptophans (W14, W41, and W52) to maintain the fold. Although there is similarity between the two structures, negative charged electrostatic surface potential in the concave face of the molecule of box 5 exhibits great difference compared to that of box 1 and other HMG boxes with known structures. That surface is involved in DNA binding. Besides, in positions which are involved in intercalating into a DNA base pair, there are hydrophobic residues in box 1 and other HMG boxes but polar residues in box 5. These differences may contribute to the loss of the DNA binding ability of box 5. PMID- 12590580 TI - Understanding copper trafficking in bacteria: interaction between the copper transport protein CopZ and the N-terminal domain of the copper ATPase CopA from Bacillus subtilis. AB - In this paper the interaction of cytoplasmic CopZ and the N-terminal domain of the CopA ATPase from Bacillus subtilis has been studied by NMR through (15)N-(1)H HSQC experiments in order to understand the role of the two proteins in the whole copper trafficking mechanism of the bacteria. It appears that the two proteins interact in a fashion similar to that of the yeast homologue proteins [Arnesano, F., Banci, L., Bertini, I., Cantini, F., Ciofi-Baffoni, S., Huffman, D. L., and O'Halloran, T. V. (2001) J. Biol. Chem. 276, 41365-41376], although the surface potentials are reversed. A structural model for the interaction is proposed. (15)N mobility studies on the free proteins and on their complex are also reported. From these data, it appears that copper is largely transferred from CopZ to CopA, thus suggesting their possible involvement in a detoxification process. Comparing functional data of homologous proteins of other bacteria, it can be concluded that this class of proteins is involved in copper homeostasis but the specific roles are species dependent. PMID- 12590581 TI - A dynamic structure for the acyl-enzyme species of the antibiotic aztreonam with the Citrobacter freundii beta-lactamase revealed by infrared spectroscopy and molecular dynamics simulations. AB - Infrared difference spectra show that at least 4 conformations coexist for the ester carbonyl group of the stable acyl-enzyme species formed between the antibiotic aztreonam and the class C beta-lactamase from Citrobacter freundii. A novel method for the assignment of the bands that arise from the ester carbonyl group has been employed. This has made use of the finding that the infrared absorption intensity of aliphatic esters is surprisingly constant, so a direct comparison with simple model esters has been possible. This has allowed a clear distinction to be made between ester and amide (protein) absorptions. The polarity of the conformer environment varies from hexane-like to strongly hydrogen-bonded. We assume that the conformer with the lowest frequency (1,690 cm(-)(1)) and hence the strongest hydrogen-bonding is the singular conformer observed in the X-ray crystallographic structure, since a good interaction via two hydrogen bonds with the oxyanion hole is seen. Molecular dynamics simulation by the method of locally enhanced sampling revealed that the motion of the ester carbonyl of the acyl-enzyme species in and out of the oxyanion hole is facile. The simulation revealed two pathways for this motion that would go through intermediates that first break one or the other of the two hydrogen bonds to the oxyanion hole, prior to departure of the carbonyl moiety out of the active site. It is likely that such motion for the acyl-enzyme species might also occur with more typical beta-lactam substrates for beta-lactamases, but their detection in the more rapid time scale may prove a challenge. PMID- 12590582 TI - Entropic nature of the interaction between promoter bound CRP mutants and RNA polymerase. AB - The interaction between CRP, T127L, S128A, and CRP and RNA polymerase bound to a 104 bp synthetic promoter were determined by ITC at 298 K and ranges from a deltaG(b) degrees = 1.4 +/- 0.8 kJ mol(-)(1) (cAMP-ligated S128A) to 4.5 +/- 0.3 kJ mol(-)(1) (cAMP-ligated double mutant CRP) with endothermicities that range from 4 +/- 3 kJ mol(-)(1) (cAMP-ligated CRP) to 47 +/- 8 kJ mol(-)(1) (cGMP ligated T127L). The interaction is, thus, entropically driven, exhibits enthalpy entropy compensation, and increases the binding affinity of the RNA polymerase to the promoter by factors ranging from 1.7 +/- 0.1 (cAMP-ligated S128A) to 6.1 +/- 0.1 (cAMP-ligated CRP). Although the binding affinities to the promoter alone, except for cAMP-ligated S128A, are the same as to a shorter 40 bp duplex containing the same CRP consensus binding site sequence (conDNA), the binding enthalpies of CRP/mutant to the promoter are lower by factors of 2-3 x than the corresponding binding enthalpies to conDNA. Small angle neutron scattering measurements on the DNA-CRP/mutant complexes in D(2)O/H(2)O solutions exhibit an increase in the Rg of the CRP/mutant component from 22 to 27-31 A that can be attributed to a conformational change in the N-terminal domain of CRP. The Rg = 27 A for the bound conDNA can be attributed to a slight unwinding of the DNA in solution that would also enhance the activation of transcription. The Rg = 53 +/- 3 A for the bound promoter is attributed to bending of the promoter in solution that can be responsible for the lower CRP/mutant-promoter binding endothermicities. PMID- 12590583 TI - Structure of tetrahydrobiopterin tunes its electron transfer to the heme-dioxy intermediate in nitric oxide synthase. AB - How 6R-tetrahydrobiopterin (H(4)B) participates in Arg hydroxylation as catalyzed by the nitric oxide synthases (NOSs) is a topic of current interest. Previous work with the oxygenase domain of inducible NOS (iNOSoxy) demonstrated that H(4)B radical formation is kinetically coupled to disappearance of an initial heme dioxy intermediate and to Arg hydroxylation in a single turnover reaction run at 10 degrees C [Wei, C.-C., Wang, Z.-Q., Wang, Q., Meade, A. L., Hemann, C., Hille, R., and Stuehr, D. J. (2001) J. Biol. Chem. 276, 315-319]. Here we used 5-methyl H(4)B to investigate how pterin structure influences radical formation and associated catalytic steps. In the presence of Arg, the heme-dioxy intermediate in 5-methyl-H(4)B-bound iNOSoxy reacted at a rate of 35 s(-)(1), which is 3-fold faster than with H(4)B. This was coupled to a faster rate of 5-methyl-H(4)B radical formation (40 vs 12.5 s(-)(1)) and to a faster and more productive Arg hydroxylation. The EPR spectrum of the enzyme-bound 5-methyl-H(4)B radical had different hyperfine structure than the bound H(4)B radical and exhibited a 3-fold longer half-life after its formation. A crystal structure of 5-methyl-H(4)B-bound iNOSoxy revealed that there are minimal changes in conformation of the bound pterin or in its interactions with the protein as compared to H(4)B. Together, we conclude the following: (1) The rate of heme-dioxy reduction is linked to pterin radical formation and is sensitive to pterin structure. (2) Faster heme-dioxy reduction increases the efficiency of Arg hydroxylation but still remains rate limiting for the reaction. (3) The 5-methyl group influences heme-dioxy reduction by altering the electronic properties of the pterin rather than changing protein structure or interactions. (4) Faster electron transfer from 5-methyl-H(4)B may be due to increased radical stability afforded by the N-5 methyl group. PMID- 12590584 TI - Zinc- and pH-dependent conformational transition in a putative interdomain linker region of the influenza virus matrix protein M1. AB - The matrix protein M1 of influenza A virus forms a shell beneath the viral envelope and sustains the virion architecture by interacting with other viral components. A structural change of M1 upon acidification of the virion interior in an early stage of virus infection is considered to be a key step to virus uncoating. We examined the structure of a 28-mer peptide (M1Lnk) representing a putative linker region between the N- and C-terminal domains of M1 by using circular dichroism, Raman, and absorption spectroscopy. M1Lnk assumes an alpha helical structure in a mildly hydrophobic environment irrespective of pH, being consistent with the X-ray crystal structures of an N-terminal fragment of M1 at pH 7 and 4. In the presence of Zn(2+), on the other hand, M1Lnk takes a partially unfolded conformation at neutral pH with a tetrahedral coordination of two Cys residues and two His residues to a Zn(2+) ion in the central part of the peptide. Upon acidification, the peptide releases the Zn(2+) ion and refolds into the alpha-helix-rich structure with a midpoint of transition at pH 5.9. The pH dependent conformational transition of M1Lnk strongly suggests that the interdomain linker region of M1 also undergoes a pH-dependent unfolding-refolding transition in the presence of Zn(2+). A small but significant portion of the M1 protein is bound to Zn(2+) in the virion, and the Zn(2+)-bound M1 molecule may play a special role in virus uncoating by changing the disposition of the N- and C-terminal domains upon acidification of the virion interior. PMID- 12590585 TI - Kinetic and docking studies of the interaction of quinones with the quinone reductase active site. AB - NAD(P)H/quinone acceptor oxidoreductase type 1 (QR1) protects cells from cytotoxic and neoplastic effects of quinones though two-electron reduction. Kinetic experiments, docking, and binding affinity calculations were performed on a series of structurally varied quinone substrates. A good correlation between calculated and measured binding affinities from kinetic determinations was obtained. The experimental and theoretical studies independently support a model in which quinones (with one to three fused aromatic rings) bind in the QR1 active site utilizing a pi-stacking interaction with the isoalloxazine ring of the FAD cofactor. PMID- 12590586 TI - An opsin mutant with increased thermal stability. AB - This report describes the biochemical characterization of a double mutant of rhodopsin (N2C,D282C) in which Cys residues engineered into the protein at positions 2 (in the amino-terminal extracellular domain) and 282 (in the extracellular loop between transmembrane helices 6 and 7) are shown to form a disulfide bond and increase the thermal stability of the unliganded or opsin form of the protein. Wild-type opsin does not survive detergent solubilization and purification at pH 7.5 and 25 degrees C. In contrast, the N2C,D282C mutant opsin survives the purification protocol and loses less than 50% activity after incubation for 20 days under the same conditions. Less than 5% is lost after 20 days at 4 degrees C. While the disulfide bond clearly has a dramatic effect on protein stability, it has a minor impact on the activity of the pigment. The MII lifetime of the mutant (6.6 min) is similar to that of the wild type (7.9 min), and the specific activity of the light-activated mutant for activation of transducin is within 20% of the wild-type activity. Therefore, it seems likely that the disulfide bond does not perturb greatly the structure of the protein. For these reasons, we anticipate that the mutant may be of use in detailed kinetic and mechanistic investigations of the ligand binding reaction and for crystallization trials involving recombinant rhodopsin, especially the unliganded opsin form of the protein. PMID- 12590587 TI - Slow binding of retinal to rhodopsin mutants G90D and T94D. AB - In an examination of the effect of three rhodopsin night blindness mutations on the rate of association of 11-cis-retinal with opsin, one of the mutations (G90D) was found to slow the rate of reaction by more than 80-fold. This effect does not appear to be general to night blindness mutations as the two other mutants (A292E and T94I) were not found to bind retinal with slowed kinetics. However, T94D was similar to G90D in that the rate of retinal binding was dramatically slowed. Gly90 and Thr94 are both located in the active site of the protein close to the Schiff base counterion Glu113. Thus, the slow kinetics of Schiff base formation appear to correlate with the introduction of a negative charge close to the Schiff base counterion, suggesting a possible role for Glu113 as a catalytic base in this reaction. Consistent with this model, the E113Q mutant was also found to bind retinal more slowly than the wild type. PMID- 12590588 TI - Characterization of rhodopsin congenital night blindness mutant T94I. AB - The Thr94 --> Ile mutation in the second transmembrane segment of rhodopsin has been reported to be associated with a congenital night blindness phenotype in a large Irish pedigree. Previously, two other known rhodopsin mutants that cause congenital night blindness, A292E and G90D, have been shown in vitro to constitutively activate the G protein transducin in the absence of a chromophore. The proposed mechanism of constitutive activation of these two mutants is an electrostatic disruption of the active site salt bridge between Glu113 and Lys296 that contributes to stabilization of the protein in the inactive state. Here, the T94I rhodopsin mutant is characterized and compared to the two other known rhodopsin night blindness mutants. The T94I mutant opsin is shown also to constitutively activate transducin. The T94I mutant pigment (with a bound 11-cis retinal chromophore), like the other known rhodopsin night blindness mutants, is not active in the dark and has wild-type activity upon exposure to light. Similar to the Gly90 --> Asp substitution, position 94 is close enough to the Schiff base nitrogen that an Asp at this position can functionally substitute for the Glu113 counterion. However, in contrast to the other night blindness mutants, the T94I MII intermediate decays with a half-life that is approximately 8-fold slower than in the wild-type MII intermediate. Thus, the one phenotype shared by all congenital night blindness mutants that is different from the wild-type protein is constitutive activation of the apoprotein. PMID- 12590589 TI - B-side charge separation in bacterial photosynthetic reaction centers: nanosecond time scale electron transfer from HB- to QB. AB - We report time-resolved optical measurements of the primary electron transfer reactions in Rhodobacter capsulatus reaction centers (RCs) having four mutations: Phe(L181) --> Tyr, Tyr(M208) --> Phe, Leu(M212) --> His, and Trp(M250) --> Val (denoted YFHV). Following direct excitation of the bacteriochlorophyll dimer (P) to its lowest excited singlet state P, electron transfer to the B-side bacteriopheophytin (H(B)) gives P(+)H(B)(-) in approximately 30% yield. When the secondary quinone (Q(B)) site is fully occupied, P(+)H(B)(-) decays with a time constant estimated to be in the range of 1.5-3 ns. In the presence of excess terbutryn, a competitive inhibitor of Q(B) binding, the observed lifetime of P(+)H(B)(-) is noticeably longer and is estimated to be in the range of 4-8 ns. On the basis of these values, the rate constant for P(+)H(B)(-) --> P(+)Q(B)(-) electron transfer is calculated to be between approximately (2 ns)(-)(1) and approximately (12 ns)(-)(1), making it at least an order of magnitude smaller than the rate constant of approximately (200 ps)(-)(1) for electron transfer between the corresponding A-side cofactors (P(+)H(A)(-) --> P(+)Q(A)(-)). Structural and energetic factors associated with electron transfer to Q(B) compared to Q(A) are discussed. Comparison of the P(+)H(B)(-) lifetimes in the presence and absence of terbutryn indicates that the ultimate (i.e., quantum) yield of P(+)Q(B)(-) formation relative to P is 10-25% in the YFHV RC. PMID- 12590590 TI - The function of the chloride ion in photosynthetic oxygen evolution. AB - The involvement of Cl(-) and several other monovalent anions in photosynthetic oxygen evolution was studied using photosystem II membranes depleted of Cl(-) by dialysis. The results of these studies differ significantly from results obtained using other depletion methods. Binding studies with glycerol as a cryoprotectant confirm our previous observations with sucrose of two interconvertible binding states of photosystem II with similar activities and with slow or fast exchange, respectively, of the bound ion. With glycerol, Cl(-) depletion decreased the oxygen evolution rate to 55% of that with Cl(-) present without decreasing the quantum efficiency of the reaction, supporting our previous conclusion that oxygen evolution can proceed at high rates in the absence of Cl(-). Further, after Cl(-) depletion the S(2) state multiline signal displayed the same periodic appearance with the same signal yield after consecutive laser flashes as with Cl( ) present. Br(-), I(-), and NO(3)(-), although with different capacities to reactivate oxygen evolution, also showed two binding modes. I(-) inhibited when bound in the low-affinity, fast-exchange mode but activated in the high-affinity mode. A comparison of the EPR properties of the S(2) state with these anions suggests that the nature of the ion or the binding mode only has a minor influence on the environment of the manganese. In contrast, F(-) completely inhibited oxygen evolution by preventing the S(2) to S(3) transition and shifted the equilibrium between the g = 4.1 and multiline S(2) forms toward the former, which suggests a considerable perturbation of the manganese cluster. To explain these and earlier observations, we propose that the role of chloride in the water splitting mechanism is to participate together with charged amino acid side chains in a proton-relay network, which facilitates proton transfer from the manganese cluster to the medium. The structural requirements likely to be involved may explain the sensitivity of oxygen evolution to Cl(-) depletion or other perturbations. PMID- 12590591 TI - Role of hydrophobic interactions in the flavodoxin mediated electron transfer from photosystem I to ferredoxin-NADP+ reductase in Anabaena PCC 7119. AB - Hydrophobic interactions play an active role in effective complex formation between ferredoxin-NADP(+) reductase (FNR) and ferredoxin (Fd) from Anabaena, where an aromatic amino acid residue on the Fd surface (F65) and three hydrophobic residues (L76, L78, and V136) on the reductase surface have been shown to be essential for the efficient electron transfer (ET) reaction between Fd and FNR (Martinez-Julvez et al. (2001) J. Biol. Chem. 276, 27498-27510). Since in this system flavodoxin (Fld) can efficiently replace Fd in the overall ET process, we have further investigated if such hydrophobic interactions are also critical in complex stabilization and ET in the FNR/Fld association. Different ET behaviors with Fld are observed for some of the mutations made at L76, L78, and V136 of Anabaena FNR. Thus, the ET interaction with Fld is almost completely lost upon introduction of negatively charged side chains at these positions, while more conservative changes in the hydrophobic patch can influence the rates of ET to and from Fld by altering the binding constants and the midpoint redox potentials of the flavin group. Therefore, our results confirm that nonpolar residues in the region close to the FAD group in FNR participate in the establishment of interactions with Fld, which serve to orient the two flavin groups in a manner such that ET is favored. In an attempt to look for the counterpart region of the Fld surface, the effect produced by the replacement of the only two nonpolar residues on the Fld surface, I59 and I92, by a Lys has also been analyzed. The results obtained suggest that these two hydrophobic residues are not critical in the interaction and ET processes with FNR. The reactivity of these I92 and I59 Fld mutants toward the membrane-anchored photosystem I (PSI) complex was also analyzed by laser flash absorption spectroscopy. From these data, significant effects are evident, especially for the I92 position of Fld, both in the association constant for complex formation and in the electron transfer rate constant in the PSI/Fld system. PMID- 12590592 TI - The electron transfer complexes of cytochrome c peroxidase from Paracoccus denitrificans. AB - We have used microcalorimetry and analytical ultracentrifugation to test the model proposed in Pettigrew et al. [(1999) J. Biol. Chem. 274, 11383-11389] for the binding of small cytochromes to the cytochrome c peroxidase of Paracoccus denitrificans. Both methods reveal complexity in behavior due to the presence of a monomer/dimer equilibrium in the peroxidase. In the presence of either Ca(2+), or higher ionic strength, this equilibrium is shifted to the dimer. Experiments to study complex formation with redox partners were performed in the presence of Ca(2+) in order to simplify the equilibria that had to be considered. The results of isothermal titration calorimetry reveal that the enzyme can bind two molecules of horse cytochrome c with K(d) values of 0.8 microM and 2.5 microM (at 25 degrees C, pH 6.0, I = 0.026) but only one molecule of Paracoccus cytochrome c 550 with a K(d) of 2.8 microM, molar binding ratios confirmed by ultracentrifugation. For both horse cytochrome c and Paracoccus cytochrome c-550, the binding is endothermic and driven by a large entropy change, a pattern consistent with the expulsion of water molecules from the interface. For horse cytochrome c, the binding is weakened 3-fold at I = 0.046 M due to a smaller entropy change, and this is associated with an increase in enzyme turnover. In contrast, neither the binding of cytochrome c-550 nor its oxidation rate is affected by raising the ionic strength in this range. We propose that, at low ionic strength, horse cytochrome c is trapped in a nonproductive orientation on a broad capture surface of the peroxidase. PMID- 12590593 TI - Carboxylate groups on the manganese-stabilizing protein are required for efficient binding of the 24 kDa extrinsic protein to photosystem II. AB - The effects of the modification of carboxylate groups on the manganese stabilizing protein on the binding of the 24 kDa extrinsic protein to Photosystem II were investigated. Carboxylate groups on the manganese-stabilizing protein were modified with glycine methyl ester in a reaction facilitated by 1-ethyl-3-(3 dimethylaminopropyl)carbodiimide. The manganese-stabilizing protein which was modified while associated with NaCl-washed membranes could bind to calcium chloride-washed PS II membranes and reconstitute oxygen evolution in a manner similar to that observed for unmodified manganese-stabilizing protein (Frankel, L.K, Cruz, J. C. and Bricker, T. M. (1999) Biochemistry 38, 14271-14278). However, PS II membranes reconstituted with this modified protein were defective in their ability to bind the extrinsic 24 kDa protein of Photosystem II. Mapping of the sites of modification was carried out by trypsin and Staphylococcus V8 protease digestion of the modified protein and analysis by MALDI mass spectrometry. These studies indicated that the domains (1)E-(71)D, (97)D-(144)D, and (180)D-(187)E are labeled when the manganese-stabilizing protein is bound to NaCl-washed Photosystem II membranes. We hypothesize that modified carboxylates, possibly residues (1)E, (32)E, (139)E, and/or (187)E, in these domains are responsible for the altered binding affinity of the 24 kDa protein observed. PMID- 12590594 TI - Dynamics of protein and chromophore structural changes in the photocycle of photoactive yellow protein monitored by time-resolved optical rotatory dispersion. AB - The dynamics of the PYP photocycle have been studied using time-resolved optical rotatory dispersion (TRORD) spectroscopy in the visible and far-UV spectral regions to probe the changes in the chromophore configuration and the protein secondary structure, respectively. The changes in the secondary structure in PYP upon photoisomerization of the chromophore can be described by two exponential lifetimes of 2 +/- 0.8 and 650 +/- 100 ms that correspond to unfolding and refolding processes, respectively. The TRORD experiments that follow the dynamics of the chromophore report three exponential components, with lifetimes of 10 +/- 3 micros, 1.5 +/- 0.5 ms, and 515 +/- 110 ms. A comparison of the kinetic behaviors of the chromophore and protein shows that during the decay of pR(465) an initial relaxation that is localized to the chromophore hydrophobic pocket precedes the formation of the chromophore and protein structures found in pB(355). In contrast, the protein and chromophore processes occur with similar time constants during inactivation of the signaling state. PMID- 12590595 TI - (4-Hydroxyphenyl)pyruvate dioxygenase from Streptomyces avermitilis: the basis for ordered substrate addition. AB - (4-hydroxyphenyl)pyruvate dioxygenase (HPPD) catalyzes the second step in the pathway for the catabolism of tyrosine, the conversion of (4 hydroxyphenyl)pyruvate (HPP) to homogentisate (HG). This reaction involves decarboxylation, substituent migration, and aromatic oxygenation. HPPD is a member of the alpha-keto acid dependent oxygenases that require Fe(II) and an alpha-keto acid substrate to oxygenate an organic molecule. We have examined the binding of ligands to HPPD from Streptomyces avermitilis. Our data show that HPP binds to the apoenzyme and that the apo-HPPD.HPP complex does not bind Fe(II) to generate active holoenzyme. The binding of HPP, phenylpyruvate (PPA), and pyruvate to the holoenzyme produces a weak ligand charge-transfer band at approximately 500 nm that is indicative of bidentate binding of the 1-carboxylate and 2-keto pyruvate oxygen atoms to the active site metal ion. For HPPD from this organism the 4-hydroxyl group of (4-hydroxyphenyl)pyruvate is a requirement for catalysis; no turnover is observed in the presence of phenylpyruvate. The rate constant for the dissociation of Fe(II) from the holoenzyme is 0.0006 s(-)(1) and indicates that this phenomenon is not significantly relevant in steady-state turnover. The addition of HPP and molecular oxygen to the holoenzyme is formally random. The basis of the ordered bi bi steady-state kinetic mechanism previously observed by Rundgren (Rundgren, M. (1977) J. Biol. Chem. 252, 5094-9) is the 3600 fold increase in oxygen reactivity when holo-HPPD is in complex with HPP. This complex reacts with molecular oxygen with a second-order rate constant of 1.4 x 10(5) M(-)(1) s(-)(1) inducing the formation of an intermediate that decays at the catalytically relevant rate of 7.8 s(-)(1). PMID- 12590596 TI - Characterization of metal ligand mutants of tyrosine hydroxylase: insights into the plasticity of a 2-histidine-1-carboxylate triad. AB - The amino acid ligands to the active site iron in the aromatic amino acid hydroxylase tyrosine hydroxylase are two histidines and a glutamate. This 2 histidine-1-carboxylate motif has been found in a number of other metalloenzymes which catalyze a variety of oxygenase reactions. As a probe of the plasticity of this metal binding site, each of the ligands in TyrH has been mutated to glutamine, glutamate, or histidine. The H336E and H336Q enzymes show dramatic decreases in iron affinity but retain substantial activity for both tyrosine hydroxylation and tetrahydropterin oxidation. The H331E enzyme shows a lesser decrease in iron affinity and is unable to hydroxylate tyrosine. Instead, this enzyme oxidizes tetrahydropterin in the absence of added tyrosine. The E376H enzyme has no significant activity, while the E376Q enzyme hydroxylates tyrosine at about 0.4% the wild-type rate. When dopamine is bound to either the H336Q or H331E enzymes, the position of the long wavelength charge-transfer absorbance band is consistent with the change in the metal ligand. In contrast, the H336E enzyme does not form a stable binary complex with dopamine, while the E376H and E376Q enzymes catalyze dopamine oxidation. PMID- 12590597 TI - Reaction of 1-amino-2-methylenecyclopropane-1-carboxylate with 1 aminocyclopropane-1-carboxylate deaminase: analysis and mechanistic implications. AB - 1-aminocyclopropane-1-carboxylate (ACC) deaminase is a pyridoxal 5'-phosphate (PLP) dependent enzyme which catalyzes the opening of the cyclopropane ring of ACC to give alpha-ketobutyric acid and ammonia. In an early study of this unusual C(alpha)-C(beta) ring cleavage reaction, 1-amino-2-methylenecyclopropane-1 carboxylic acid (2-methylene-ACC) was shown to be an irreversible inhibitor of ACC deaminase. The sole turnover product was identified as 3-methyl-2-oxobutenoic acid. These results provided strong evidence supporting the ring cleavage of ACC via a nucleophilic addition initiated process, thus establishing an unprecedented mechanism of coenzyme B(6) dependent catalysis. To gain further insight into this inactivation, tritiated 2-methylene-ACC was prepared and used to trap the critical enzyme nucleophiles. Our results revealed that inactivation resulted in the modification of an active site residue, Ser-78. However, an additional 5 equiv of inhibitor was also found to be incorporated into the inactivated enzyme after prolonged incubation. In addition to Ser-78, other nucleophilic residues modified include Lys-26, Cys-41, Cys-162, and Lys-245. The location of the remaining unidentified nucleophile has been narrowed down to be one of the residues between 150 and 180. Labeling at sites outside of the active site is not enzyme catalyzed and may be a consequence of the inherent reactivity of 2 methylene-ACC. Further experiments showed that Ser-78 is responsible for abstracting the alpha-H from d-vinylglycine and may serve as the base to remove the beta-H in the catalysis of ACC. However, it is also likely that Ser-78 serves as the active site nucleophile that attacks the cyclopropane ring and initiates the fragmentation of ACC, while the conserved Lys-51 is the base required for beta-H abstraction. Clearly, the cleavage of ACC to alpha-ketobutyrate by ACC deaminase represents an intriguing conversion beyond the common scope entailed by coenzyme B(6) dependent catalysts. PMID- 12590598 TI - Potentiometric analysis of UDP-galactopyranose mutase: stabilization of the flavosemiquinone by substrate. AB - UDP-galactopyranose mutase is a flavoprotein which catalyses the interconversion of UDP-galactopyranose and UDP-galactofuranose. The enzyme is of interest because it provides the activated biosynthetic precursor of galactofuranose, a key cell wall component of many bacterial pathogens. The reaction mechanism of this mutase is intriguing because the anomeric oxygen forms a glycosidic bond, which means that the reaction must proceed by a novel mechanism involving ring breakage and closure. The structure of the enzyme is known, but the mechanism, although speculated on, is not resolved. The overall reaction is electrically neutral but a crypto-redox reaction is suggested by the requirement that the flavin must adopt the reduced form for activity. Herein we report a thermodynamic analysis of the enzyme's flavin cofactor with the objective of defining the system and setting parameters for possible reaction schemes. The analysis shows that the neutral semiquinone (FADH(*)) is stabilized in the presence of substrate and the fully reduced flavin is the anionic FADH(-) rather than the neutral FADH(2). The anionic FADH(-) has the potential to act as a rapid 1-electron donor/acceptor without being slowed by a coupled proton transfer and is therefore an ideal crypto-redox cofactor. PMID- 12590599 TI - Distinct effects on heparan sulfate structure by different active site mutations in NDST-1. AB - Heparan sulfate polymerization and modification take place in the Golgi compartment. The modification reactions are initiated by glucosaminyl N deacetylase/N-sulfotransferase (NDST), a bifunctional enzyme that removes N acetyl groups from selected N-acetyl-d-glucosamine units followed by N-sulfation of the generated free amino groups. Four isoforms of NDST have been identified. NDST-1 and -2 have a wide and largely overlapping tissue distribution, but it is not known if they can act on the same heparan sulfate chain. We have introduced point mutations into NDST-1 cDNA, which selectively destroy the N-deacetylase or N-sulfotransferase activity of the enzyme [Wei, Z., and Swiedler, S. J. (1999) J. Biol. Chem. 274, 1966-70 and Sueyoshi, T., et al. (1998) FEBS Lett. 433, 211-4]. Stable 293 cell lines expressing the NDST-1 mutants were then generated. Structural analyses of heparan sulfate synthesized by these cells and by cells overexpressing wild-type NDST-1 demonstrate that the N-deacetylation step is not only prerequisite but also rate-limiting, determining the degree of N-sulfation. Transfection of mutant NDST-1 lacking N-deacetylase activity had no effect on heparan sulfate sulfation, while cells expressing wild-type enzyme or NDST-1 lacking N-sulfotransferase activity both resulted in the production of oversulfated heparan sulfate. Since no increase in the amount of N-unsubstituted glucosamine residues was seen after transfection of the mutant lacking N sulfotransferase activity, the results also suggest that two different enzyme molecules can act on the same glucosamine unit. In addition, we show that oversulfation of heparan sulfate produced by cells tranfected with wild-type NDST 1 or the mutant lacking N-sulfotranferase activity results in decreased sulfation of chondroitin sulfate. PMID- 12590600 TI - Arginine vasopressin inhibition of cyclin D1 gene expression blocks the cell cycle and cell proliferation in the mouse Y1 adrenocortical tumor cell line. AB - Arginine vasopressin (AVP) is a nonapeptide long known as an endocrine and paracrine regulator of important systemic functions, namely, vasoconstriction, gluconeogenesis, corticosteroidogenesis, and excretion of water and urea. Here we report, for the first time, that AVP specifically inhibits expression of the cyclin D1 gene, leading to cell cycle blockage and halting cell proliferation. In G0/G1-arrested mouse Y1 adrenocortical tumor cells, maintained in serum-free medium (SFM), AVP mimics FGF2, promoting rapid ERK1/2 activation (5 min) followed by c-Fos protein induction (2 h). PKC inhibitor Go6983 and PI3K inhibitors wortmannin and LY294002 all inhibit ERK1/2 activation by AVP, but not by FGF2. Thus, AVP and FGF2 concur to activate ERK1/2 by different regulatory pathways. However, AVP is not a mitogenic factor for Y1 cells. On the contrary, AVP strongly antagonizes FGF2 late induction (2-5 h) of the cyclin D1 gene, down regulating both cyclin D1 mRNA and protein. AVP inhibition of cyclin D1 expression is sufficient to block G1 phase progression and cell entry into the S phase, monitored by BrdU nuclear labeling. In addition, AVP completely inhibits proliferation of Y1 cells in 10% fetal calf serum (10% FCS) medium. On the other hand, ectopic expression of the cyclin D1 protein renders Y1 cells resistant to AVP for both entry into the S phase in SFM and continuous proliferation in 10% FCS medium. In conclusion, inhibition of cyclin D1 expression by AVP is an efficient mechanism of cell cycle blockage and consequent proliferation inhibition in Y1 adrenocortical cells. PMID- 12590601 TI - Concerning the chemical nature of tubulin subunits that cap and stabilize microtubules. AB - There is no definitive evidence on the nature of the cap at microtubule ends that is responsible for dynamic instability behavior. It was, therefore, of interest that steady-state microtubules assembled in 20 mM P(i) buffer and pulsed for 15 60 min with [gamma-(32)P]GTP contained approximately 26 [(32)P]P(i)/microtubule [Panda et al. (2002) Biochemistry 41, 1609-1617]. It was concluded that microtubules are capped with a tubulin-GDP-P(i) subunit at the end of each its 13 protofilaments and that this is responsible for stabilizing microtubules in the growth phase. Also, because microtubules with [(32)P]P(i) were isolated despite the presence of 20 mM P(i), it was concluded that P(i) in terminal tubulin-GDP P(i) subunits does not exchange with solvent. These observations are inconsistent with our finding that tubulin-GDP-P(i) subunits do not stabilize microtubules and with evidence that the nucleotide, and presumably also P(i), in subunits at microtubule ends exchanges with solvent. We have resolved this discrepancy by finding that during the pulse period the added [(32)P]GTP was almost quantitatively hydrolyzed. The so-formed [(32)P]P(i) labeled the 20 mM P(i) buffer, and this exchanged into tubulin-GDP subunits in the core of the microtubule. Evidence for this was our finding of virtually identical [(32)P]P(i) in microtubules pulsed with [(32)P]GTP with a specific activity that varied 11 fold by using either 100 or 1,100 microM GTP in the reaction. Label uptake was insensitive to the [(32)P]GTP specific activity because in both cases hydrolysis generated 20 mM [(32)P]P(i) with a virtually identical specific activity. Also, approximately 0.4 mol of [(32)P]P(i) /tubulin dimer was found in microtubules when steady-state microtubules in 20 mM P(i) were pulsed with a trace amount of [(32)P]P(i). This stoichiometry is consistent with a 25 mM K(d) previously reported for P(i) binding to tubulin-GDP subunits in microtubules. It is concluded that, under the conditions used for the [(32)P]GTP pulse labeling, (32)P was incorporated into the entire microtubule from [(32)P]P(i) released into the solution, rather than into a tubulin-GDP-P(i) cap, from [(32)P]GTP. Thus, there is no evidence that tubulin-GDP-P(i) subunits accumulate in and stabilize microtubule ends. PMID- 12590602 TI - Multiple metalloproteinases process protransforming growth factor-alpha (proTGF alpha). AB - Shedding of TNF-alpha requires a single cleavage event, whereas the ectodomain of proTGF-alpha is cleaved at N-proximal (N-terminal) and membrane proximal (C terminal) sites to release mature TGF-alpha. Tumor necrosis factor-alpha converting enzyme (TACE) was shown to have a central role in the shedding of both factors. Here we show that cleavage of the proTGF-alpha C-terminal site, required for release of mature growth factor, is less sensitive to a panel of hydroxamates than TNF-alpha processing. Recombinant TACE cleaves TNF-alpha and N-terminal TGF alpha peptides 50-fold more efficiently than the C-terminal TGF-alpha peptide. Moreover, fractionation of rat liver epithelial cell membranes yields two populations: one contains TACE and cleaves peptides corresponding to TNF-alpha and both proTGF-alpha processing sites, while the other lacks detectable TACE and cleaves only the C-terminal proTGF-alpha processing site. Activities in both fractions are inhibited by hydroxamates and EDTA but not by cysteine, aspartate, or serine protease inhibitors. Both membrane fractions also contain ADAM 10. ADAM 10 correctly cleaves peptides and a soluble form of precursor TGF-alpha (proTGFecto) at the N-terminal site but not the C-terminal site. However, the kinetics of N-terminal peptide cleavage by ADAM 10 are 90-fold less efficient than TACE. Our findings indicate that while TACE is an efficient proTGF-alpha N terminal convertase, a different activity, distinguishable from TACE, exists that can process proTGF-alpha at the C-terminal site. A model that accounts for these findings and the requirement for TACE in TGF-alpha shedding is proposed. PMID- 12590603 TI - Evaluation of a modular strategy for the construction of novel polydactyl zinc finger DNA-binding proteins. AB - In previous studies, we have developed a technology for the rapid construction of novel DNA-binding proteins with the potential to recognize any unique site in a given genome. This technology relies on the modular assembly of modified zinc finger DNA-binding domains, each of which recognizes a three bp subsite of DNA. A complete set of 64 domains would provide comprehensive recognition of any desired DNA sequence, and new proteins could be assembled by any laboratory in a matter of hours. However, a critical parameter for this approach is the extent to which each domain functions as an independent, modular unit, without influence or dependence on its neighboring domains. We therefore examined the detailed binding behavior of several modularly assembled polydactyl zinc finger proteins. We first demonstrated that 80 modularly assembled 3-finger proteins can recognize their DNA target with very high specificity using a multitarget ELISA-based specificity assay. A more detailed analysis of DNA binding specificity for eight 3-finger proteins and two 6-finger proteins was performed using a target site selection assay. Results showed that the specificity of these proteins was as good or better than that of zinc finger proteins constructed using methods that allow for interdependency. In some cases, near perfect specificity was achieved. Complications due to target site overlap were found to be restricted to only one particular amino acid interaction (involving an aspartate in position 2 of the alpha-helix) that occurs in a minority of cases. As this is the first report of target site selection for designed, well characterized 6-finger proteins, unique insights are discussed concerning the relationship of protein length and specificity. These results have important implications for the design of proteins that can recognize extended DNA sequences, as well as provide insights into the general rules of recognition for naturally occurring zinc finger proteins. PMID- 12590604 TI - The interaction of DOF transcription factors with nucleosomes depends on the positioning of the binding site and is facilitated by maize HMGB5. AB - The expression of genes involved in C(4) photosynthesis in maize is under tight tissue-specific and light-dependent control. There is strong evidence that this control is at least in part brought about by DOF transcription factors binding to the respective promoters. We analyzed the interaction of DOF1 and DOF2 proteins with a functional and a cryptic endogenous binding site derived from the maize phosphoenolpyruvate carboxylase promoter (-300 bp region) in the nucleosomal context. Various DNA fragments comprising this promoter region were reconstituted into mononucleosomes from purified components, resulting in different positions of the DOF binding sites on the nucleosome surface. Binding of recombinant transcription factors to the different types of nucleosomes was examined using electrophoretic mobility shift assays. Changing the translational position of the binding site on the nucleosome surface strongly affected the efficiency of the interaction with the DOF factors. Deletion of individual recognition motifs revealed a positive impact of DOF protein binding to the main binding site on interactions with the cryptic binding site. The addition of the chromosomal high mobility group (HMG) protein HMGB5 to the binding reaction mixture facilitated nucleosome binding of the transcription factor independent from the position of the recognition sites. The relevance of the data for the activation of the promoter in vivo is discussed. PMID- 12590605 TI - Factors that contribute to efficient catalytic activity of a small Ca2+-dependent deoxyribozyme in relation to its RNA cleavage function. AB - Recently, we found a small Ca(2+)-dependent deoxyribozyme (unmodified), d(GCCTGGCAG(1)G(2)C(3)T(4)A(5)C(6)A(7)A(8)C(9)G(10)A(11)GTCCCT), with cleavage activity for its RNA substrate, r(AGGGACA downward arrow UGCCAGGC) ( downward arrow denotes the RNA cleavage site), in the presence of Ca(2+) and developed a functional SPR sensor chip with this deoxyribozyme [Okumoto, Y., Ohmichi, T., and Sugimoto, N. (2002) Biochemistry 41, 2769-2773]. In the study presented here, to clarify the factors contributing to the efficient catalytic activity of the unmodified deoxyribozyme, RNA cleavage reactions were carried out using 24 mutant deoxyribozymes containing one unnatural DNA nucleotide, such as dI (2' deoxyinosine), 7-deaza-dG, 2-aminopurine, 7-deaza-dA, 2-amino-dA, dm(5)C (5 methyl-2'-deoxycytosine), or d(P)C (5-propynyl-2'-deoxycytosine). The K(m) values (Michaelis constants) with the mutants that lacked N7 and O6 of G(1) and O6 of G(2) were 4.5 and 6.6 times that of the unmodified one, respectively. The k(cat) value (cleavage rate constant) with the mutants that lacked O6 of G(10) was 0.025 times that of the unmodified one. The results of UV melting curves, SPR kinetics, and CD spectra supported the quantitative idea that the catalytic activity of the unmodified form was achieved using Ca(2+). On the basis of these results, a preliminary model for two G(1) x A(8) and G(2) x A(7) mismatched base pairs such as G(anti) x A(anti) formed in the catalytic loop is proposed. The factor of 10 increase in the k(cat)/K(m) value of the mutant deoxyribozyme, which has C(9) substituted with d(P)C, suggests that the base stacking interaction between the substituted propynyl group in dC and the nearest-neighbor base grew stronger. Thus, substituting d(P)C for dC in the catalytic loop would be one of the best ways to increase the catalytic activity of the deoxyribozyme. PMID- 12590606 TI - Stimulation of DNA strand slippage synthesis by a bulge binding synthetic agent. AB - It has been postulated that bulged structures may be intermediates in the DNA strand slippage synthesis associated with the expansion of nucleotide repeats in various neurodegenerative diseases and cancer. To probe the possible role of bulged structures in this process, we have synthesized a wedge-shaped spirocyclic molecule, DDI (double-decker intercalator), on the basis of our earlier work with the bulge-specific derivative prepared from the enediyne antitumor antibiotic neocarzinostatin chromophore. Using a series of primers/templates containing nucleotide repeats [(AAT)(3)/(ATT)(5), (ATT)(3)/(AAT)(5), (CAG)(3)/(CTG)(5), (CA)(4)C/(GT)(7)G, (GT)(4)G/(CA)(7)C, T(9)/A(30), T(20)/A(30)] with the Klenow fragment of Escherichia coli DNA polymerase I, we find that DDI markedly enhances the formation of long DNA products, whose synthesis would require strand slippage to occur. DDI-induced slippage synthesis is more pronounced as the incubation proceeds and at limiting enzyme levels. The gel band pattern of the synthesized DNA products reflects the particular nucleotide repeat unit and is not altered by DDI. The lack of any drug effect on primer extension on M13 DNA and heteropolymeric 62-mer templates, where strand slippage is much less likely to occur, suggests that stimulation of slippage synthesis by DDI is not due to a direct effect on the enzyme. By contrast, other DNA-binding agents, such as ethidium bromide, distamycin, and doxorubicin, inhibit the formation of slippage induced DNA products, but this block can be overcome by DDI, presumably by its destabilizing duplex DNA-binding sites for these other agents. We propose that DDI binds to or induces the formation of a bulge or related structure, which promotes DNA strand slippage and its consequent expansion of nucleotide repeats during replication by DNA polymerase I and that this action provides insight into the development of agents that interfere with nucleotide expansions found in various disease states. PMID- 12590607 TI - Effect of pH on the iso-1-cytochrome c denatured state: changing constraints due to heme ligation. AB - The effect of pH on the denatured state (3 M guanidine hydrochloride) was evaluated with fluorescence spectroscopy for four variants of iso-1-cytochrome c, AcTM (no surface histidines), AcH26 (surface histidine at position 26), AcH54 (surface histidine at position 54), and AcH54I52 (stabilizing I52 mutation added to AcH54). Changes in the compactness and the heme ligation of the denatured state, as a function of pH, were monitored through changes in Trp 59-heme fluorescence quenching. With the AcTM and AcH26 variants, no change in the fluorescence intensity occurs from pH 4 to 10. However, for the AcH54 and AcH54I52 variants the fluorescence intensity drops significantly between pH 4 and 6, consistent with His 54 binding to the heme of cytochrome c. Between pH 8 and 10 fluorescence intensity increases again, indicating that the His 54 is displaced from the heme. The data are consistent with lysines 4 and 5 being the primary heme ligands at alkaline pH, under denaturing conditions. This conclusion was confirmed by site-directed mutagenesis. Thermodynamic analysis indicates that heme-ligand affinity in the denatured state is controlled primarily by sequence position (loop size) and that when histidines are present they inhibit lysine ligation until approximately pH 8.5-9.0 as compared to pH 7.5 with the AcTM variant. Thus, at physiological pH, histidine ligands provide the primary constraint on the denatured state of cytochrome c. The heme-Trp 59 distance in the denatured state of iso-1-cytochrome c, derived from analysis by Forster energy transfer theory, is approximately 26 A at pH 4 and 10, much shorter than the random coil prediction of 56 A. Surprisingly, the heme-Trp 59 distance in the His 54 bound conformation only drops to approximately 21 A, consistent with an extended conformation for the short polypeptide segment separating heme and Trp 59. PMID- 12590608 TI - Catalytic and regulatory domains of doublecortin kinase-1. AB - Doublecortin kinase-1 (DCK1) is a newly described multidomain protein kinase with a sequence significantly similar to those of both CaM kinases (CaMKs) and doublecortin, the product of the gene mutated in X-linked lissencephaly/double cortex syndrome, a severe developmental disorder of the nervous system. Functional studies have revealed microtubule binding and polymerization activities of the doublecortin domain, yet little is known regarding the enzymatic properties and regulation of the kinase catalytic domain. We have identified and report here notable similarities as well as differences between the catalytic and regulatory properties of DCK1 and those of the CaMKs. Using synthetic peptide substrates modeled on synapsin I, a substrate recognition motif for DCK1 of Hyd-Arg-Arg-X-X-Ser/Thr-Hyd was derived. The similarity of this motif to that of CaMKI [Lee, J. C., Kwon, Y.-G., Lawrence, D. S., and Edelman, A. M. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 6413-6417] is consistent with the 59% level of amino acid sequence similarity between their catalytic domains. DCK1 catalytic activity is enhanced by mutagenic introduction of negative charge at Thr-239, a residue in a position equivalent to that of Thr-177 of CaMKI, the activation loop site for regulation by CaM kinase kinase. Unlike CaMKs, DCK1 is not directly activated by Ca(2+)-bound CaM. However, truncation of a pseudosubstrate-like sequence in the C-terminus of DCK1 results in an approximately 6-fold enhancement of activity. Thus, DCK1 demonstrates the potential to be regulated by relief of autoinhibition in response to signal(s) distinct from Ca(2+)-bound CaM and potentially by activation loop phosphorylation and to phosphorylate intracellular targets at sites similar to those recognized by CaMK pathways. PMID- 12590609 TI - Solution X-ray scattering data show structural differences among chimeras of yeast and chicken calmodulin: implications for structure and function. AB - We present here the first evidence, obtained by the use of small-angle X-ray scattering, of the solution structures of chimeras constructed from yeast (Saccharomyces cerevisiae, Sc) and chicken (Gallus gallus, Gg) calmodulin (CaM). The chimeric proteins used in this study are Sc(1-129)/Gg(130-148), Sc(1 128)/Gg(129-148), Sc(1-87)/Gg(88-148), and Sc(1-72)/Gg(73-148) CaMs, in which Sc(1-)(n)() and Gg(()(n)(+1)-148) descend from yeast and chicken CaM in the chimeric proteins, respectively. Under the Ca(2+)-saturated condition, the solution structure of Sc(1-128)/Gg(129-148) CaM has a dumbbell-like shape which is characteristic of vertebrate-type CaM, while that of Sc(1-129)/Gg(130-148) CaM takes an intermediate structure between the dumbbell-like shape and a compact globular shape. The results provide the direct evidence that the replacement of Asp(129) with Ser(129) induces an interaction between two lobes of Sc(1 129)/Gg(130-148) CaM and brings them close together. It implies that a site interacting with the N-lobe is induced in the C-lobe, although site IV that is unable to bind Ca(2+) hinders the ability of the C-lobe to undergo the conformational change to the full open state. In the presence of both Ca(2+) and a peptide synthesized to mimic the CaM binding domain on myosin light chain kinase, MLCK-22p, the solution structures of these chimeric CaMs take a similar compact globular shape but their interactions are quite different. The solution structure and interactions of Sc(1-72)/Gg(73-148) CaM are similar to those of Sc(1-87)/Gg(88-148) CaM. The structure of Sc(1-87)/Gg(88-148) CaM is similar to that of Sc(1-128)/Gg(129-148) CaM, but their interactions are different. The result indicates that the replacement of Glu(119) with Ala(119) has a critical effect on their interactions. Thus, the functional differences among these chimeric CaMs, which have been reported previously [Nakashima, K., et al. (1996) Biochemistry 35, 5602-5610], have been interpreted on the basis of the structures and interactions. PMID- 12590610 TI - Thermal and urea-induced unfolding of the marginally stable lac repressor DNA binding domain: a model system for analysis of solute effects on protein processes. AB - Thermodynamic and structural evidence indicates that the DNA binding domains of lac repressor (lacI) exhibit significant conformational adaptability in operator binding, and that the marginally stable helix-turn-helix (HTH) recognition element is greatly stabilized by operator binding. Here we use circular dichroism at 222 nm to quantify the thermodynamics of the urea- and thermally induced unfolding of the marginally stable lacI HTH. Van't Hoff analysis of the two-state unfolding data, highly accurate because of the large transition breadth and experimental access to the temperature of maximum stability (T(S); 6-10 degrees C), yields standard-state thermodynamic functions (deltaG(o)(obs), deltaH(o)(obs), deltaS(o)(obs), deltaC(o)(P,obs)) over the temperature range 4-40 degrees C and urea concentration range 0 /=100 times faster than EH(4) did. Conversely, EH(4) reduced 2,6-dimethyl-1,4-benzoquinone and molecular oxygen 90 and 40 times faster than EH(2), respectively. Comparison of the rates of reduction of the above substrates by EH(2) and EH(4) with their corresponding steady-state kinetic parameters for kinetic competence leads to the conclusion that reduction of lipoyl substrates occurs with EH(2) while reduction of diaphorase substrates occurs with EH(4). PMID- 12590612 TI - Retinyl esters are the substrate for isomerohydrolase. AB - Regeneration of 11-cis retinal from all-trans retinol in the retinal pigment epithelium (RPE) is a critical step in the visual cycle. The enzyme(s) involved in this isomerization process has not been identified and both all-trans retinol and all-trans retinyl esters have been proposed as the substrate. This study is to determine the substrate of the isomerase enzyme or enzymatic complex. Incubation of bovine RPE microsomes with all-trans [(3)H]-retinol generated both retinyl esters and 11-cis retinol. Inhibition of lecithin retinol acyltransferase (LRAT) with 10-N-acetamidodecyl chloromethyl ketone (AcDCMK) or cellular retinol binding protein I (CRBP) diminished the generation of both retinyl esters and 11 cis retinol from all-trans retinol. The 11-cis retinol production correlated with the retinyl ester levels, but not with the all-trans retinol levels in the reaction mixture. When retinyl esters were allowed to form prior to the addition of the LRAT inhibitors, a significant amount of isomerization product was generated. Incubation of all-trans [(3)H]-retinyl palmitate with RPE microsomes generated 11-cis retinol without any detectable production of all-trans retinol. The RPE65 knockout (Rpe65(-/-)) mouse eyecup lacks the isomerase activity, but LRAT activity remains the same as that in the wild-type (WT) mice. Retinyl esters in WT mice plateau at 8 weeks-of-age, but Rpe65(-/-) mice continue to accumulate retinyl esters with age (e.g., at 36 weeks, the levels are 20x that of WT). Our data indicate that the retinyl esters are the substrate of the isomerization reaction. PMID- 12590613 TI - Nardilysin cleaves peptides at monobasic sites. AB - Nardilysin (N-arginine dibasic convertase, EC 3.4.24.61) was first identified on the basis of its ability to cleave peptides containing an arginine dibasic pair, i.e., Arg-Arg or Arg-Lys. However, it was observed that an aromatic residue adjacent to the dibasic pair (i.e., Phe-Arg-Lys) could alter the cleavage site. In this study we determined whether nardilysin can cleave peptides at a single basic residue. Nardilysin cleaves beta-endorphin at the monobasic site, Phe(17) Lys(18), with a k(cat)/K(m) of 2 x 10(8) M(-)(1) min(-)(1). This can be compared to a k(cat)/K(m) of 8.5 x 10(8) M(-)(1) min(-)(1) for cleavage between a dibasic pair in dynorphin B-13. Nardilysin also cleaves calcitonin at His-Arg and somatostatin-14 at Cys-Lys. We examined the hydrolysis of fluorogenic peptides based on the beta-endorphin 12-24 sequence, Abz-T-P-L-V-T-L-X(1)-X(2)-N-A-I-I-K-Q EDDnp. Nardilysin hydrolyzes the peptides when X(1)-X(2) = F-K, F-R, W-K, M-K, Y K, and L-K. The kinetics of cleavage at F-K and F-R are similar; however, K-F is not hydrolyzed. Nardilysin cleaves at two monobasic sites M-K and F-R of the kallidin model peptide Abz-MISLMKRPPGFSPFRSSRI-NH(2), releasing desArg(10) kallidin (KRPPGFSPF). However, nardilysin does not release desArg(10) kallidin from the physiological precursor low molecular weight kininogen. These studies extend the range of potential substrates for nardilysin and further substantiate that nardilysin is a true peptidase. PMID- 12590614 TI - Cytoplasmic truncation of glycoprotein Ib alpha weakens its interaction with von Willebrand factor and impairs cell adhesion. AB - The interaction of the platelet glycoprotein (GP) Ib-IX-V complex with von Willebrand factor (VWF) is a critical step in the adhesion of platelets to the subendothelial matrix following endothelial cell damage, particularly under arterial flow conditions. In the human GP Ib-IX-V complex, the recognition of VWF appears to be mediated entirely by GP Ibalpha, the largest of four GP Ib-IX-V polypeptides. The goal of the present study was to investigate the involvement of the cytoplasmic domain of GP Ibalpha in the GP Ib-IX-VWF interaction under both static conditions and in the presence of high fluid shear stress. Using Chinese hamster ovary (CHO) cells that express GP Ibbeta, GP IX, and either wild-type GP Ibalpha or GP Ibalpha mutants missing various lengths of the cytoplasmic domain, we evaluated adhesion and flow-driven cell rolling on immobilized VWF in a parallel-plate flow chamber. Cells expressing GP Ibalpha polypeptides with truncations of 6-82 amino acids rolled faster than cells expressing wild-type GP Ibalpha. Cells that expressed polypeptides with intact actin-binding protein 280 binding sites (truncated to residue 582 of 610) rolled more slowly than those expressing GP Ibalpha with longer truncations. The rolling velocity of cells expressing truncated GP Ibalpha mutants increased with decreasing VWF coating density. In addition, a fraction of the truncated cells exhibited saltatory translocation at the lower VWF densities. Studies measuring the GP Ibalpha-VWF bond strength of three of the mutants using laser tweezers showed that progressive deletion of the cytoplasmic domain led to progressive weakening of the strength of individual GP Ibalpha-VWF bonds. PMID- 12590615 TI - Tau polymerization: role of the amino terminus. AB - The abnormal polymerization of the tau molecule into insoluble filaments is a seminal event in the neurodegenerative process underlying Alzheimer's disease. Previous experimentation has shown that the microtubule-binding repeat region of the molecule is vital for its ability to polymerize in vitro into filaments similar to those found in Alzheimer's disease. However, it is becoming clear that regions outside the microtubule-binding repeat, such as exons 2 and 3 and the carboxy-terminal tail, can greatly influence its polymerization. Since it has been previously postulated that the amino terminus of tau could be involved in generating pathological conformations in the disease state, its role in the polymerization process was investigated. This report demonstrates that the removal of the amino terminus greatly inhibits the polymerization of the tau molecule, reducing both the rate and extent of polymerization. These results support the hypothesis that the ability of tau to form specific conformations involving the amino terminus is an early event in the formation of tau polymers in the disease state. Furthermore, the mutation of arginine 5 to leucine ((R)5(L)), mimicking an amino-terminal tau mutation found in a single case of FTDP-17, enhances the polymerization of the tau molecule. Therefore, the amino terminus of the tau molecule, while largely overlooked in studies of its polymerization, is a significant contributor to the polymerization process. PMID- 12590617 TI - Neurotransmission of cognition, part 1, Dopamine is a hitchhiker in frontal cortex: norepinephrine transporters regulate dopamine. AB - ISSUE: Since the frontal cortex has a low density of dopamine transporters, dopamine has to be inactivated there by hitching a ride on the norepinephrine transporter of neighboring norepinephrine neurons. PMID- 12590618 TI - Clinical correlates of inpatient suicide. AB - BACKGROUND: Previous suicide assessment research has led to standard predictors of risk. Despite this, there are approximately 30,000 suicides per year in the United States, 5% to 6% of which occur in hospitals. The primary purpose of this study is to improve our ability to assess risk and intervene successfully. METHOD: Charts from 76 patients who committed suicide while in the hospital, or immediately after discharge, were reviewed. The week before suicide was rated for both standard risk predictors and, using items from the Schedule for Affective Disorders and Schizophrenia (SADS), for presence and severity of symptoms found to be correlated with acute risk in recent studies. RESULTS: Regarding standard predictors, only 49% (N = 37) had any prior suicide attempt and 25% (N = 19) were admitted for this reason. Thirty-nine percent (30/76) were admitted for suicidal ideation, but 78% denied suicidal ideation at their last communication about this; 46% (N = 35) showed no evidence of psychosis; of those on precautions (N = 45), 51% (N = 23) were on q 15 minute suicide checks or 1:1 observation; and 28% (N = 21) had a no-suicide contract in effect. On SADS ratings, 79% (N = 60) met criteria for severe or extreme anxiety and/or agitation. CONCLUSION: Standard risk assessments and standard precautions used were of limited value in protecting this group from suicide. Adding severity of anxiety and agitation to our current assessments may help identify patients at acute risk and suggest effective treatment interventions. The importance of a matched comparison group to ascertain if this sample can be blindly discriminated from inpatients who do not commit suicide is clear. PMID- 12590619 TI - Antidepressants and driver impairment: empirical evidence from a standard on-the road test. AB - BACKGROUND AND METHOD: The current review summarizes the major results from all published studies from 1983 to 2000 (9 double-blind, crossover, placebo controlled studies in healthy volunteers and 1 double-blind, baseline-controlled study in patients) that have determined the effects of antidepressants on actual driving performance using a standard test. That test measures driving impairment from vehicular "weaving" (i.e., standard deviation of lateral position [SDLP]) during 1 hour of on-the-road driving in normal traffic. RESULTS: Changes in SDLP after acute doses of sedating antidepressants (i.e., amitriptyline, imipramine, doxepin, and mianserin) were comparable to those seen in drivers conducting the same test with a blood alcohol concentration of 0.8 mg/mL or more. Driving performance of subjects returned to placebo levels after 1 week of treatment, except after treatment with mianserin, for which the impairing effect lasted unabated over treatment. Nocturnal doses of sedating antidepressants (i.e., dothiepin, mianserin, and mirtazapine), however, did not produce residual driving impairment when measured the next day. Nonsedating antidepressants (i.e., moclobemide, fluoxetine, paroxetine, venlafaxine, and nefazodone) generally did not affect SDLP. However, SDLP rose to unacceptable levels after administration of combinations of nonsedating antidepressants and benzodiazepines with incompatible pharmacokinetic profiles. Correlational analyses demonstrated that conventional tests of psychomotor performance or self-ratings of side effects did not strongly predict antidepressant effects on SDLP. Regression analysis revealed a strong linear relation between antidepressant effects in the standard driving test and the number of patients reporting somnolence in clinical trials with the same antidepressants. CONCLUSION: Application of actual driving tests remains essential to conclusively defining the potential hazard of drugs for driving. PMID- 12590620 TI - Prior antipsychotic prescribing in patients currently receiving clozapine: a case note review. AB - BACKGROUND: Clozapine is indicated for the treatment of resistant schizophrenia, which is usually defined as failure to respond to adequate trials of 2 antipsychotics. It is thought that only clozapine is likely to be effective in such cases and that other drugs are ineffective. We sought to discover prior patterns of antipsychotic prescribing in schizophrenic patients eventually prescribed clozapine. METHOD: Prescribing histories were obtained from prescription charts and case notes for all inpatients prescribed clozapine in 4 hospitals in southeast London during April 2001. RESULTS: 120 patients were identified, of whom 112 had been diagnosed with schizophrenia or schizoaffective disorder and whose data were analyzed. The mean duration of illness was 15.1 years. Subjects had experienced a mean of 9.2 (range, 2-35) episodes of antipsychotic prescription before clozapine was first used, with 5.7 (range, 0 25) episodes constituting adequate trials (drug used at therapeutic dose for 6 weeks). The mean number of different antipsychotics used was 5.5 (range, 1-13), with a mean of 4.0 (range, 0-12) given an adequate trial. Ninety percent of patients (N = 101) had received an atypical antipsychotic before first use of clozapine, and 65% (N = 73) had previously received antipsychotic polypharmacy. The mean maximum theoretical delay in using clozapine was 5.0 years (range, 0 11.1 years). Longer delay was significantly (p <.05) associated with being aged over 30 years at the time of the study, being diagnosed with psychotic illness before the introduction of clozapine, and completing adequate trials of 2 different antipsychotics before the introduction of clozapine or risperidone. CONCLUSION: Clozapine treatment was quite likely delayed for longer than is clinically desirable. This delay may have important effects on quality of life, clinical outcome, and health resource utilization. PMID- 12590621 TI - Nortriptyline for treatment-resistant depression. AB - BACKGROUND: Up to 30% of patients with major depression fail to respond to an antidepressant trial, with most taking a selective serotonin reuptake inhibitor (SSRI) as initial treatment. While the tricyclic antidepressants might be effective for SSRI nonresponders, they have been relegated to third- and fourth line treatment. This study assesses the efficacy of nortriptyline for patients with treatment-resistant major depression. METHOD: 92 patients with treatment resistant DSM-III-R major depression, with resistance defined by at least 1, but no more than 5, well-documented adequate trials of antidepressants during the current episode, were treated openly with nortriptyline for 6 weeks. Patients were titrated up to full target doses of nortriptyline within 1 week, with target blood levels of 100 ng/mL. Response was defined as a 50% or greater decrease of baseline 17-item Hamilton Rating Scale for Depression score. We performed an intent-to-treat analysis with the last observation carried forward. RESULTS: Approximately 40% of patients were responders (N = 39) and 12% were remitters (N = 11) after 6 weeks of nortriptyline. Over a third of patients were unable to complete the trial. CONCLUSION: Nortriptyline was effective for over a third of patients with treatment-resistant depression, and nortriptyline should be considered as potential treatment if patients fail to respond to other antidepressants. PMID- 12590622 TI - Evaluating the efficacy of habit reversal: comparison with a placebo control. AB - BACKGROUND: The purpose of this study was to compare the effectiveness of habit reversal with a placebo control as a treatment for chronic nail biting in adults. METHOD: Thirty adults with a chronic nail-biting problem (occurring > or = 5 times/day nearly every day for > or = 4 weeks and causing physical damage or social impairment) were randomly assigned to a placebo control or habit reversal group. Five participants withdrew from the study prior to the completion of treatment. The remaining individuals in both groups received a total of 2 hours of treatment over 3 sessions. Individuals in the habit reversal group (N = 13) received the components of awareness training, competing response training, and social support. Individuals in the placebo control group (N = 12) simply discussed their nail biting. At pretreatment, posttreatment, and a 5-month follow up, nail length was measured, photographs were taken of the damaged nails and later rated by independent observers, and data on participant depression, anxiety, and self-esteem were obtained. Treatment compliance and acceptability data were collected at posttreatment only. RESULTS: Results showed that habit reversal produced a greater increase in nail length at posttreatment and follow up when compared with the placebo. Data from the independent raters confirmed these findings. Habit reversal was also viewed as a more acceptable intervention by the participants. At posttreatment, the habit reversal group had increased their nail length by 22% from pretreatment compared with a 3% increase for the placebo group. At follow-up, the habit reversal group maintained a 19% increase in nail length from pretreatment compared with a 0% increase for the placebo group. CONCLUSION: Findings from this study suggest habit reversal is more effective than a placebo control and should be considered a well-established intervention for body-focused repetitive behaviors. PMID- 12590623 TI - An open-label, flexible-dose study of olanzapine in the treatment of trichotillomania. AB - BACKGROUND: Thus far, only selective serotonin reuptake inhibitors have been systematically studied in the treatment of trichotillomania, and the results are conflicting. This open-label study is the first to systematically evaluate an atypical neuroleptic, olanzapine, as a monotherapy in the treatment of trichotillomania. METHOD: Twenty-one patients were screened and 18 patients were enrolled in a 3-month open-label study of olanzapine for trichotillomania (diagnosis based on modified DSM-IV criteria). Patients with comorbid psychiatric disorders or on treatment with psychoactive medication were excluded. Olanzapine was titrated gradually in 2.5-mg/week increments up to a maximum dose of 10 mg/day. RESULTS: Seventeen patients who completed at least 1 week of olanzapine treatment were evaluated. Hair pulling, as measured by the Massachusetts General Hospital Hairpulling Scale, decreased by 66% from baseline (p < or =.001), and mean scores on the Hamilton Rating Scale for Anxiety decreased by 63% (p < or =.05). Clinical Global Impressions scale scores also revealed significant improvement as a whole (p < or =.001), with 4 patients having complete symptom remission at the end of the study period. CONCLUSION: Findings suggest that olanzapine may be an effective monotherapy for trichotillomania. PMID- 12590624 TI - Screening for bipolar disorder in the community. AB - BACKGROUND: Our goal was to estimate the rate of positive screens for bipolar I and bipolar II disorders in the general population of the United States. METHOD: The Mood Disorder Questionnaire (MDQ), a validated screening instrument for bipolar I and II disorders, was sent to a sample of 127,800 people selected to represent the U.S. adult population by demographic variables. 85,358 subjects (66.8% response rate) that were 18 years of age or above returned the survey and had usable data. Of the nonrespondents, 3404 subjects matched demographically to the 2000 U.S. Census data completed a telephone interview to estimate nonresponse bias. RESULTS: The overall positive screen rate for bipolar I and II disorders, weighted to match the 2000 U.S. Census demographics, was 3.4%. When adjusted for the nonresponse bias, the rate rose to 3.7%. Only 19.8% of the individuals with positive screens for bipolar I or II disorders reported that they had previously received a diagnosis of bipolar disorder from a physician, whereas 31.2% reported receiving a diagnosis of unipolar depression. An additional 49.0% reported receiving no diagnosis of either bipolar disorder or unipolar depression. Positive screens were more frequent in young adults and low income households. The rates of migraine, allergies, asthma, and alcohol and drug abuse were substantially higher among those with positive screens. CONCLUSION: The positive MDQ screen rate of 3.7% suggests that nearly 4% of American adults may suffer from bipolar I and II disorders. Young adults and individuals with lower income are at greater risk for this largely underdiagnosed disorder. PMID- 12590625 TI - Weight, lipids, glucose, and behavioral measures with ziprasidone treatment in a population with mental retardation. AB - BACKGROUND: Atypical antipsychotics effectively reduce maladaptive behavior in individuals with mental retardation, yet bring significant weight gain and metabolic anomalies. Ziprasidone, a weight-neutral antipsychotic in patients with schizophrenia or schizoaffective disorder, has not been studied in a population with mental retardation and maladaptive behaviors. METHOD: Forty patients with mental retardation and maladaptive behaviors who had gained excessive weight or were inadequately responsive to other agents were switched to ziprasidone. Weight, total cholesterol, HDL, LDL, triglycerides, and frequency of maladaptive behavior were recorded at baseline and after 6 months of ziprasidone treatment. RESULTS: Ziprasidone treatment was associated with a significant weight loss of 8.1 lb (3.6 kg) as well as a significant reduction in total cholesterol and triglycerides (p < or =.05). The monthly frequency of the maladaptive behavior remained unchanged or improved in 72% (18/25) of the patients in whom maladaptive behavior was assessed. CONCLUSION: Ziprasidone effectively reduces the frequency of maladaptive behavior in a patient group with mental retardation without causing weight gain or metabolic disturbances. PMID- 12590626 TI - Randomized controlled trial of different models of care for nursing home residents with dementia complicated by depression or psychosis. AB - OBJECTIVE: To compare the outcomes of 3 interventions for the management of dementia complicated by depression or psychosis: psychogeriatric case management, general practitioners with specialist psychogeriatric consultation, and standard care for nursing home residents. METHOD: The sample for this 12-week randomized controlled trial consisted of 86 subjects with dementia from 11 Sydney, Australia, nursing homes, of whom 34 had depression, 33 had depression and psychosis, and 19 had psychosis. All participants received full psychiatric assessments and physical examinations. Information was obtained from the residents' families and nursing home staff. Depression measures included the Even Briefer Assessment Scale for Depression, Hamilton Rating Scale for Depression, Cornell Scale for Depression in Dementia, and Geriatric Depression Scale. Psychosis measures included the Behavioral Pathology in Alzheimer's Disease Rating Scale, Neuropsychiatric Inventory, and Scale for the Assessment of Positive Symptoms. Data were obtained from nursing home records on prescription of psychotropic medication and demographic information. Management plans were formulated by a multidisciplinary team before random assignment to interventions. RESULTS: All 3 groups improved from pretreatment to posttreatment on depression scales for depression groups and psychosis scales for psychosis groups. Mode of management appeared to make no difference in rate or amount of improvement; neither of the treatment group-by-time interactions were significant. Neither use of antidepressants nor use of antipsychotics predicted depression or psychosis outcomes. CONCLUSION: Participation in the study was associated with improvement in depression and psychosis, perhaps because of the presence of a psychogeriatric team, the increased attention focused on residents, or the generalization of active intervention techniques to control subjects. A formula-driven psychogeriatric team case management approach was not significantly more effective than a consultative approach or standard care. This study demonstrates the difficulties and feasibility of conducting service-oriented research in nursing homes. PMID- 12590627 TI - The pharmacokinetics of sertraline excretion into human breast milk: determinants of infant serum concentrations. AB - BACKGROUND: The purpose of this study was to attain a new landmark in the area of selective serotonin reuptake inhibitor therapy during lactation by establishing a basis for interpreting infant serum concentrations and for minimizing infant exposure in the absence of treatment-emergent side effects. METHOD: Breast milk and paired maternal and infant sera were collected following maternal treatment with sertraline monotherapy (25-200 mg/day) administered once daily. Sertraline and its major metabolite were measured in breast milk and serum samples using high-performance liquid chromatography with UV detection (limit of detection = 2 ng/mL). RESULTS: Twenty-six nursing women with DSM-IV major depressive disorder participated in the study; the mean (SD) daily sertraline dose was 123.9 (62.8) mg/day. Fifteen women submitted 182 breast milk samples for analysis of gradient (foremilk to hindmilk) and time course of medication excretion. The milk/plasma ratio was highly variable (range, 0.42-4.81). A significant gradient and time course of excretion for both sertraline (p <.001 for both) and desmethylsertraline (p <.001 for gradient and p <.046 for time course) were observed, with the highest concentrations observed in the hindmilk 8 to 9 hours after maternal ingestion. Mathematical modeling of sertraline and desmethylsertraline excretion revealed that discarding breast milk 9 hours after maternal dose decreased the infant daily dose of sertraline by a mean of 17.1% (1.8%). Twenty-two mother/infant sera pairs were obtained. Sertraline was detectable in 4 infants (18% of sample), and desmethylsertraline was found in 11 infants (50% of sample). The mean (SD) maximum calculated nursing infant dose of sertraline, 0.67 (0.61) mg/day, and desmethylsertraline, 1.44 (1.36) mg/day, represented 0.54% (0.49%) of the maternal daily dose. The maximum infant dose of desmethylsertraline (p <.002) significantly correlated with infant serum desmethylsertraline concentrations (ng/mL). In contrast, maternal daily dose, duration of medication exposure, and infant age and weight at sampling did not correlate with either detectability (< 2 ng/mL vs. > or = 2 ng/mL) or absolute concentrations (ng/mL) in infant serum. No adverse events were reported or documented in any infant. CONCLUSION: These results extend previous studies by demonstrating the utility of breast milk analysis in interpreting infant serum concentrations and minimizing infant exposure. PMID- 12590628 TI - Impulsive aggressive behavior: open-label treatment with citalopram. AB - BACKGROUND: Results from open-label and placebo-controlled trials suggest that the selective serotonin reuptake inhibitors reduce impulsive aggressive behavior. The objective of this open-label study was to investigate whether citalopram treatment has anti-aggressive effect on impulsive aggressive subjects meeting DSM IV criteria for a cluster B personality disorder or intermittent explosive disorder. METHOD: In this 8-week trial, subjects were initiated on 20 mg/day of citalopram and titrated up to 60 mg/day by the fourth week, if tolerated. The primary outcome measure was the Overt Aggression Scale-Modified (OAS-M), a scale used to quantify verbal and physical aggression, subjective irritability, and overt irritability. Secondary outcome measures included the Barratt Impulsiveness Scale and Buss-Durkee Hostility Inventory. RESULTS: Of 25 subjects enrolled, 20 completed the study. The mean daily dose was 45.5 mg, and citalopram was generally well tolerated. Statistically significant decreases were found in the OAS-M aggression scores (32.82 +/- 19.76 to 4.73 +/- 7.57, p =.000), subjective irritability scores (3.50 +/- 0.60 to 1.45 +/- 1.18, p =.000), and overt irritability scores (3.23 +/- 0.81 to 0.91 +/- 1.02, p =.000). CONCLUSION: These results suggest that citalopram is an effective treatment for reducing impulsive aggressive behavior. PMID- 12590629 TI - The Maudsley Bipolar Disorder Project: the effect of medication, family history, and duration of illness on IQ and memory in bipolar I disorder. AB - BACKGROUND: Despite the growing recognition of the importance of cognitive impairment in psychiatric disorders, the effect of clinical factors, such as medication use and family history of affective disorders, on cognition in bipolar I disorder patients still remains unclear. This study examines the contribution of known and potential predictors to both general intellectual function and memory in a representative population of bipolar I disorder patients. METHOD: Of the 425 patients receiving treatment within a defined catchment area, 63 were identified as having bipolar I disorder. Of these patients, 43 were enrolled in the study and participated in a personal interview by a psychiatrist. All patients were invited to participate in a personal interview by a psychiatrist, and information on family history, past psychiatric history, past and current treatments, duration of illness, and age at onset was collected, in addition to demographic data. Cognitive performance was assessed using the Wechsler Adult Intelligence Scale-Revised, the National Adult Reading Test, and the Wechsler Memory Test III. RESULTS: Forty-three patients with DSM-IV bipolar I disorder were enrolled into the Maudsley Bipolar Disorder Project. Patients on treatment with antipsychotic drugs had a lower current full scale IQ, lower general memory scores, and lower working memory scores. A family history of affective disorders was associated with a higher full scale IQ, but not with either general or working memory measures. Duration of illness was negatively associated with general memory scores, but had no effect on either IQ or working memory measures. CONCLUSION: Current antipsychotic medication, duration of illness, and family history of affective disorder were the most significant predictors of IQ and memory function in bipolar I disorder patients. PMID- 12590630 TI - Combination therapy: is clinical practice leading the way? PMID- 12590632 TI - Analgesic effects of antidepressants. PMID- 12590634 TI - Prolonged erections associated with ziprasidone treatment: a case report. PMID- 12590635 TI - A polymorphism in the IGF-I gene influences the age-related decline in circulating total IGF-I levels. AB - OBJECTIVE: Recent studies have demonstrated an association between a 192 bp polymorphism of the IGF-I gene and total IGF-I serum levels, birth weight, body height and the risk of developing diabetes and cardiovascular diseases later on in life. This IGF-I gene polymorphism in the promoter region of the IGF-I gene may directly influence the expression of IGF-I. In the present study we evaluated the role of this polymorphism in the age-related decline in serum IGF-I levels. SUBJECTS AND METHODS: All subjects were participants of the Rotterdam Study, a population-based cohort study of diseases in the elderly. We studied a total group of 346 subjects, who comprised two subgroups: a randomly selected population-based sample of 196 subjects, and a group of 150 subjects selected on IGF-I genotype. In the total group of 346 individuals the relationship between this 192 bp polymorphism and the age-related decline in circulating total IGF-I levels was studied. RESULTS: Homozygous carriers of the 192 bp allele demonstrated significant decline in serum IGF-I with age (r=-0.29, P=0.002). This decline is similar to that seen in the general population. An age-related decline in serum total IGF-I was not observed in heterozygotes (r=-0.06, P=0.48) and non carriers (r = -0.12, P=0.32). Interestingly, the relationship between age and serum IGF-binding protein-3 levels showed the same pattern. CONCLUSION: We observed only in homozygous carriers of the 192 bp alleles of the IGF-I gene an age-related decline in circulating total IGF-I levels, but not in heterozygotes and non-carriers of the 192 bp allele. We hypothesize that this IGF-I gene polymorphism directly or indirectly influences GH-mediated regulation of IGF-I secretion. PMID- 12590636 TI - Augmented serum levels of the IGF-I/IGF-binding protein-3 ratio in pre-menopausal patients with type I breast cysts. AB - OBJECTIVE: Gross cystic disease (GCD) is the most common benign breast pathology. Although breast cysts are not considered pre-malignant lesions, an increased risk of breast cancer has been reported for patients with type I cysts (Na(+)/K(+)<3). Furthermore, an augmented IGF-I/IGF-binding protein-3 (IGFBP-3) ratio has been described in breast cancer patients. The objective was to evaluate serum IGF-I and binding protein concentrations of type I and type II cyst patients as compared with healthy women. METHODS: Twenty-four patients with type I cysts, 17 with type II cysts and 25 healthy women were evaluated. Serum IGF-I, IGFBP-3 and IGFBP-1 concentrations were measured by IRMA. RESULTS: IGF-I concentrations were significantly higher in sera from patients with type I cysts than in patients with type II cysts. A highly significant decrease of IGFBP-3, the major IGFBP, was found in patients with type I cysts with respect to healthy women, whereas no significant difference was evident between the different cyst types. The IGF I/IGFBP-3 ratio, an estimate of biologically active IGF-I, was very significantly higher in patients with type I cysts than in both type II patients and healthy women. IGFBP-1 levels were significantly lower in patients with type I than in controls and type II cysts. The IGF-I/IGFBP-1 ratio was significantly higher in patients with type I cysts than in type II bearers and healthy women. Estrogen levels correlated with IGF-I in patients and controls. CONCLUSIONS: The enhanced levels of IGF-I/IGFBP-3 found in patients with type I cysts could eventually be associated with the increased risk of breast cancer described for this group. PMID- 12590637 TI - GH replacement therapy increases plasma osteoprotegerin levels in GH-deficient adults. AB - OBJECTIVE: Osteoprotegerin (OPG), a glycoprotein belonging to the tumor necrosis factor receptor family, is an endogenous inhibitor of osteoclastogenesis produced by cells of the osteoblast lineage. OPG is a key cytokine involved in the regulation of osteoblast/osteoclast cross-talk. Since GH replacement therapy in GH deficiency (GHD) activates bone remodeling and increases bone mass, we investigated if short-term GH replacement therapy affects plasma OPG levels. DESIGN AND METHODS: Eighteen adults with GHD, ranging from 17 to 51 Years (nine childhood-onset and nine adult-onset) were enrolled in the study. All subjects were on stable replacement therapy, especially sex hormones. The starting dose of GH replacement therapy was 4 microg/kg per day x 7 days/week, and was progressively increased according to the serum IGF-I values. Biochemical parameters of bone and mineral metabolism were measured before and after 6 Months of GH replacement therapy. Bone mass density (BMD) was monitored at three skeletal sites (lumbar vertebrae, femur, radius) by dual-energy X-ray absorptiometry. RESULTS: After 6 Months of therapy, ionized calcium, parathyroid hormone and 25-OH vitamin D did not change, whereas total serum calcium and urinary calcium excretion increased significantly (P<0.01). Also osteocalcin and urinary deoxypyridinoline/24 h increased significantly (P<0.02, P<0.05 respectively). Mean basal T-scores of BMD values showed an osteopenic state, which remained unchanged after GH therapy. Plasma OPG increased significantly after 6 Months of therapy (P<0.02) and this increase was significantly correlated with the increase of osteocalcin (r=-0.52; P=0.04) and deoxypyridinoline values (r=-0.64; P=0.011). CONCLUSIONS: Our results suggest that the bone anabolic effect of GH replacement therapy could in part be mediated by a positive bone balance at each remodeling unit due to the inhibitory action of OPG on osteoclastogenesis. PMID- 12590638 TI - Impact of successful transsphenoidal surgery on cardiovascular risk factors in acromegaly. AB - BACKGROUND: Cardiac abnormalities develop in patients with acromegaly as a consequence of effects of GH/IGF-I on the heart and related cardiovascular risk factors. OBJECTIVE: To evaluate the possible contribution of postoperative variations in blood pressure (BP), glucose tolerance and insulin sensitivity to the cardiac improvement reported in patients who have been cured of acromegaly. DESIGN: Thirty-one patients with acromegaly were studied before and 6 Months after successful transsphenoidal surgery, defined by normal age-related IGF-I concentrations and glucose-suppressed GH concentrations <1 microg/l. METHODS: Cardiovascular parameters were assessed by Doppler echocardiography and 24-h ambulatory blood pressure monitoring. Insulin sensitivity indexes were calculated on the basis of fasting and post-load glycaemia and insulinaemia and referred to as HOMA(ISI) and OGTT(ISI), respectively. RESULTS: Successful surgery was confirmed to improve left ventricular mass index (LVMI) and diastolic filling significantly. Mean 24-h systolic BP values decreased (P=0.009) and BP rhythm was restored in 12 of 15 patients with a blunted preoperative profile. Glucose tolerance normalized in patients with preoperative glucose intolerance (n=7) or diabetes mellitus (n=3). HOMA(ISI) and OGTT(ISI) increased (P=0.0001 for each parameter), indicating a marked improvement in insulin sensitivity. The postoperative reduction in LVMI correlated with increased insulin sensitivity (P<0.001 for both indexes), but not with other parameters. Improved diastolic filling correlated with the reduction in LVMI. CONCLUSIONS: Successful surgery in patients with acromegaly induces a significant improvement in haemodynamic and metabolic risk factors. This study suggests a direct link between insulin resistance and acromegalic cardiomyopathy. PMID- 12590640 TI - Anthropometric, haemodynamic, humoral and hormonal evaluation in patients with incidental adrenocortical adenomas before and after surgery. AB - OBJECTIVE: To compare clinical and humoral parameters before and after surgery in patients with incidental adrenocortical adenomas. DESIGN: Six patients with subclinical Cushing's syndrome and nine with non-functioning adenomas were investigated before and 12 Months after removal of the mass. METHODS: Anthropometric (body weight, body mass index and waist to hip ratio), haemodynamic (blood pressure and heart rate), metabolic (lipids and oral glucose tolerance test (OGTT)), hormonal (cortisol, plasma renin activity, aldosterone, androgens and catecholamines) and bone metabolism (hydroxyproline, parathyroid hormone, osteocalcin and ostase) parameters were evaluated. RESULTS: In the whole group, a significant decrease in body weight (69.7+/-3.5 vs 70.8+/-3.5 kg, P<0.03), in systolic (135.3+/-5.1 vs 145.6+/-4.9 mmHg, P<0.009) and diastolic (83.7+/-1.9 vs 91.0+/-3.5 mmHg, P<0.03) blood pressure and in glucose levels in response to OGTT (106.4+/-9.6 vs 127.5+/-6.5 mg/dl, P<0.05) was observed after surgery. All other parameters examined did not change significantly. This trend was also found in both groups separately. Analytical data showed a high frequency of overweight/obesity (66.6%), hypertension (66.6%) and impaired glucose profile (26.6%) in our patients, with a greater prevalence of these cardiovascular risk factors in the subclinical Cushing's syndrome group. After surgery, values normalized or improved in eight out of ten hypertensive patients and in three out of four patients with impaired glucose profile. CONCLUSIONS: Solid adrenocortical incidentalomas are associated with some cardiovascular risk factors which may be corrected after removal of the mass. Therefore, surgery may be an appropriate choice in patients with subclinical Cushing's syndrome but also in those with solid non-functioning adenomas and coexistent cardiovascular risk factors. PMID- 12590639 TI - The relationship between pituitary tumour transforming gene (PTTG) expression and in vitro hormone and vascular endothelial growth factor (VEGF) secretion from human pituitary adenomas. AB - OBJECTIVE: Pituitary tumour transforming gene (PTTG) is a recently identified protooncogene, ubiquitously expressed in pituitary tumours at levels higher than those detected in normal pituitary. Although the precise function of PTTG protein is unknown, in vitro experiments have shown that it induces angiogenesis. In this study, we have examined the potential relationship between the level of PTTG expression and tumour phenotype, tumour size, in vitro pituitary hormone secretion and release of vascular endothelial growth factor (VEGF), a potent angiogenic factor. METHODS: Pituitary tumours (12 somatotroph, five lactotroph, five corticotroph and 18 non-functioning) were studied by cell culture, measuring the basal secretion of anterior pituitary hormones and VEGF in vitro. Immunocytochemistry was used to confirm the clinical diagnosis and tumour phenotype. PTTG mRNA expression was investigated by comparative RT-PCR. Tumour Volume was quantitated from pre-operative MRI scans. RESULTS: PTTG expression was significantly increased 2.7-fold in somatotroph tumours compared with non functioning adenomas (P<0.01, ANOVA). A positive correlation was demonstrated between PTTG expression and in vitro GH secretion (r=0.41, P<0.01, Spearman) but no correlations were found for any of the other pituitary hormones. In 16 out of 40 pituitary tumours, we were able to determine the in vitro secretion of VEGF and relate this to PTTG expression. All of the adenomas tested secreted measurable VEGF but there was no correlation between the amount of VEGF secreted and either the tumour phenotype or PTTG expression. Neither PTTG expression nor VEGF secretion correlated with tumour Volume. CONCLUSIONS: Our studies have confirmed the presence of PTTG in pituitary adenomas and demonstrated a higher level of expression in somatotroph tumours and a significant correlation with GH secretion. We failed to demonstrate a relationship between PTTG expression and production of the angiogenic factor, VEGF, or tumour Volume. Thus, although PTTG induces angiogenesis experimentally, it seems unlikely that a VEGF-mediated angiogenic mechanism occurs during pituitary tumour progression. PMID- 12590641 TI - Close association of severe hyponatremia with exaggerated release of arginine vasopressin in elderly subjects with secondary adrenal insufficiency. AB - OBJECTIVE: Hyponatremia occurs not infrequently in hypopituitarism. Arginine vasopressin (AVP)-induced impaired water excretion is found in patients with hypopituitarism and experimental models of glucocorticoid deficiency. DESIGN: The present study was undertaken to determine whether augmented release of AVP is involved in the development of hyponatremia in elderly subjects with secondary adrenal insufficiency. METHODS: Forty patients with ACTH-deficient, secondary adrenal insufficiency were examined. They were divided into three groups according to the age at which diagnosis was ascertained (group A <20 Years, group B 20-64 Years, and group C>or=65 Years). RESULTS: Hyponatremia was more manifest in the elderly group than in the other two groups, serum sodium (Na) levels being 124.7 mmol/l in the elderly group, a value significantly less than 141.5 and 133.5 mmol/l in groups A and B. Plasma AVP levels seemed likely to be high compared with the respective hypo-osmolality in plasma in the elderly group, as plasma AVP levels were 1.7 pmol/l despite a mean plasma osmolality of 259 mmol/kg. Such an alteration was less clear in group B and was not found in group A. Therefore, elevation of plasma AVP was apparent in the elderly patients. Hydrocortisone replacement promptly normalized serum Na levels from 125 to 142 mmol/l (P<0.01) and reduced plasma AVP levels from 1.7 to 0.9 pmol/l (P<0.05), which were comparable to the respective plasma osmolality in the elderly patients. CONCLUSION: These results indicate that non-suppressible release of AVP is crucially involved in the impaired water excretion and hyponatremia seen in elderly patients with secondary adrenal insufficiency compared with the younger patients, and that exaggerated release of AVP becomes manifest as the subjects grow older. PMID- 12590642 TI - Diurnal rhythm of free estradiol during the menstrual cycle. AB - OBJECTIVE: To investigate the diurnal rhythm of estrogens in normally cyclic women during reproductive life. DESIGN: Multiple saliva sampling in normally cyclic healthy women during reproductive life at different phases of their menstrual cycles was carried out. METHODS: Salivary estradiol was measured by radioimmunoassay in samples collected every 2 h for 24 h from 15 normally cyclic healthy women during reproductive life during the menstrual phase, the late follicular/peri-ovulation phase, the early to mid luteal phase and the late luteal phase, respectively, of their menstrual cycles. The levels of salivary estradiol were analyzed by means of periodic regression. RESULTS: A daily biological rhythm of free estradiol was found after quantification with a nonlinear periodic regression model. The observed diurnal free estradiol rhythm consists of two major components: an asymmetrically peaked diurnal cycle and ultradian harmonics in the range of 6 to 12 h. The diurnal and ultradian rhythms were remarkably consistent throughout the menstrual cycle in terms of mesor (24 h mean level), peak width and amplitude. There was a tendency for the 24-h rhythm acrophases to converge in the early morning, while the acrophase of the menstrual phase occurred significantly later than in the late follicular/peri-ovulation phase. CONCLUSIONS: The diurnal rhythm of estradiol has a similar complex temporal organization for different menstrual phases. The menstrual cycle mainly modulates the acrophase of the diurnal rhythm. PMID- 12590643 TI - Maternal serum inhibin A levels are a marker of a viable trophoblast in incomplete and complete miscarriage. AB - OBJECTIVE: From early gestation the human trophoblast secretes large amounts of inhibin A and activin A, and their measurement provides a value for predicting the outcome in women who become pregnant after assisted reproductive techniques. The aim of the study was to investigate the putative role of maternal serum inhibin A and activin A levels as markers of a viable trophoblast in women who miscarry. DESIGN: Controlled cross-sectional study. METHODS: One group consisted of 65 healthy pregnant women (controls), progressing to deliver a healthy singleton baby and another group consisted of 54 miscarriages (38 incomplete (27 non-viable, 11 anembryonic pregnancies) and 16 complete). Maternal blood samples were collected between 5 and 12 weeks of gestation. RESULTS: Serum human chorionic gonadotrophin concentrations in women with incomplete or complete miscarriages were significantly (both P<0.001) lower than in controls; activin A levels being lowest only in women with a complete miscarriage (P<0.001). On the other hand, inhibin A levels were significantly lower in incomplete or complete miscarriage than in controls (both P<0.0001). CONCLUSIONS: Maternal serum inhibin A, but not activin A, determination reflects the lack of a viable trophoblast in complete miscarriage. PMID- 12590644 TI - Somatostatin infusion withdrawal: studies in the acute and recovery phase of anorexia nervosa, and in obesity. AB - OBJECTIVE: Changes in GH/IGF-I axis activity occur in both anorexia nervosa (AN) and obesity (OB). A GH hypersecretory state with very low plasma IGF-I levels is present in AN, whereas in morbid OB, GH secretion is dull and plasma IGF-I levels are generally preserved. Endogenous GHRH activity in AN and OB has never been directly studied, although indirect evidence would indicate that GHRH function is altered in either condition, possibly enhanced and reduced respectively. Somatostatin (SS) infusion withdrawal (SSIW) is followed by a rebound rise of plasma GH in animals and humans, an event which, allegedly, is mediated by endogenous GHRH release. METHODS: In the present study, 28 young women, eight with active AN (A-AN), six with AN in the recovery phase (R-AN), eight with morbid OB, and six healthy age-matched normal weight subjects (NW), were studied. All subjects underwent, on different occasions, the following two tests: (i) acute GHRH injection (1 microg/kg, i.v.); (ii) infusion of SS (9 microg/kg per h i.v. over 60 min), with blood samples drawn prior to and at different intervals after drug injections. Plasma GH levels were measured at each time interval in all sessions, and, in addition, baseline plasma estradiol, free triiodothyronine, TSH, IGF-I and insulin were measured at -30 min. RESULTS: Baseline plasma GH concentrations were significantly higher in A-AN than in NW (4.7+/-0.7 vs 2.1+/ 0.6 microg/l, P<0.01). Baseline GH levels in R-AN were also higher than in NW, but the difference did not reach statistical significance (5.6+/-1.7 microg/l, not significant (NS)). Baseline plasma GH concentrations were significantly lower in OB than in NW (0.3+/-0.1 microg/l, P<0.01). GHRH-stimulated GH release was significantly higher in A-AN than in NW (mean change in area under the curve (DeltaAUC) 1904.9+/-626.1 vs 613.9+/-75.9 microg/l per min, P<0.01), whereas no statistically significant difference was present between R-AN and NW (mean DeltaAUC 638.2+/-293.0 microg/l per min, NS); in OB, GHRH failed to evoke a plasma GH rise (mean DeltaAUC 239.8+/-89.9 microg/l per min vs A-AN, R-AN, and NW, P<0.01). SS infusion markedly reduced plasma GH concentrations in both A-AN and R-AN and, to a lesser extent, in NW, but failed to do so in OB. In A-AN, SSIW was followed by a plasma GH rise markedly higher than that present in NW (mean DeltaAUC 193.0+/-42.3 vs 60.1+/-11.4 microg/l per min, P<0.01), whereas in R-AN the GH response after SSIW was nearly superimposable on that registered in NW (mean DeltaAUC 72.9+/-22.8 microg/l per min, NS). There were no changes in plasma GH levels after SSIW in OB (mean DeltaAUC 22.8+/-9.7 microg/l per min). In all groups, DeltaAUCs of the GH response to GHRH and after SSIW were highly positively correlated (r=0.7, P<0.01). CONCLUSIONS: These data support the view that a high endogenous GHRH tone, which subsides in the recovery phase of the disease, is present in AN, whereas GHRH hypofunction, possibly associated with pituitary impairment, might indicate OB. PMID- 12590645 TI - A prospective study of thyroid function, morphology and autoimmunity in young patients with type 1 diabetes. AB - OBJECTIVE: Thyroid autoantibodies (TA) and thyroid ultrasonography are widely used in the diagnosis of autoimmune thyroid disease (AITD). However, we know little of the significance of having ultrasonographic abnormalities (USabn) without having any other signs of AITD. In a previous population-based study of 105 young patients with type 1 diabetes (T1DM) we found a high prevalence (42%) of USabn. In the present study we evaluate the development of both USabn and TA in a 3-Year follow-up of this cohort. DESIGN: Of the 105 previously investigated children and adolescents with T1DM (aged 5-21 Years), 101 were re-examined. Serum concentrations of tri-iodothyronine (T(3)), thyroxine (T(4)), TSH, thyroid peroxidase antibodies (TPOab) and thyroglobulin antibodies (Tgab), as well as thyroid size and morphology were determined in all patients. RESULTS: During the 3 Years follow-up period, the prevalence of thyroid dysfunction increased from 5 to 8%, the prevalence of TPOab was unchanged at 13%, while the prevalence of Tgab decreased from 14 to 7%. The prevalence of USabn increased from 42 to 49%. Most patients presented USabn at both examinations. Patients with USabn had a higher prevalence of TA than those without USabn (P=0.038) and higher serum levels of TSH (P=0.027). All patients with thyroid dysfunction presented with USabn. However, many patients with USabn had no other signs of AITD. CONCLUSIONS: A high prevalence of thyroid dysfunction, TA and thyroid USabn were found in young patients with T1DM. Thyroid USabn was a sensitive but non-specific marker of AITD and is therefore unsuitable for screening purposes. Instead, we recommend regular screening using serum TSH in the follow-up of young diabetic patients. PMID- 12590646 TI - Ectopic TSH-secreting pituitary adenoma of the vomerosphenoidal junction. AB - OBJECTIVE: We describe an unusual case of ectopic TSH-secreting pituitary adenoma arising from the vomerosphenoidal junction. CLINICAL PRESENTATION: A 52-Year-old man with a long-standing history of hyperthyroidism was referred to the University Hospital in September 2001 because of increasingly disabling symptoms of nasal obstruction. For the past 18 Years the patient had complained of palpitations, hypertension, weight loss, and nervousness. A presumptive diagnosis of Graves' disease was made. Treatment with methimazole was begun, but the patient was lost to follow-up. On admission, physical examination revealed signs of hyperthyroidism and a large diffuse goiter. Tests of thyroid function showed inappropriate secretion of TSH with hyperthyroidism. Both a TSH-secreting pituitary adenoma and resistance to thyroid hormone could be taken into account. There was no evidence of pituitary tumour by magnetic resonance imaging (MRI), but a large space-occupying lesion involving the nasal cavity and the nasopharynx was incidentally discovered. INTERVENTATION AND TECHNIQUE: Using an endoscopic endonasal approach, the tumour was removed en bloc together with the sphenoid floor, sphenoid rostrum, bony septum, and part of the soft palate mucosa. Histological features and immunophenotype were those of a TSH-secreting tumour. CONCLUSION: Although exceedingly rare, ectopic TSH-secreting pituitary tumour should be borne in mind in cases of inappropriate secretion of TSH with hyperthyroidism and no evidence of pituitary tumour by computed tomography and/or MRI when a mass located along the migration path of the Rathke's pouch is demonstrated by radiological examination. To our knowledge, this is only the second reported case in the literature. PMID- 12590648 TI - Immunohistochemical detection of FGF-23 protein in tumors that cause oncogenic osteomalacia. AB - OBJECTIVE: Oncogenic hypophosphatemic osteomalacia (OOM) is a rare disease characterized by hypophosphatemia, inappropriately low levels of circulating 1,25 dihydroxyvitamin D(3) and osteomalacia. The disease is most commonly caused by benign mesenchymal tumors that produce, among several other factors, fibroblast growth factor-23 (FGF-23). Current evidence thus suggests that this protein has an important role in the regulation of phosphate homeostasis. By producing polyclonal antibodies against human FGF-23 protein we wanted to determine the localization of FGF-23 protein in OOM tumors that express FGF-23 mRNA. DESIGN AND METHODS: Three polyclonal antibodies were raised in rabbits against three different peptides with sequences derived from human FGF-23: [Cys-70]FGF-23(51 69)amide, [Tyr-223]FGF-23(206-222)amide and [Tyr-224]FGF-23(225-244)amide. One of the resulting antisera was subsequently used for immunohistochemistry on sections from five different tumors causing OOM. FGF-23 mRNA expression was confirmed with in situ hybridization. RESULTS: After affinity purification, two of three antisera detected recombinant human FGF-23 by Western blot analysis. Through immunohistochemical analysis using the anti-[Tyr-224]FGF-23(225-244)amide antibody and through in situ hybridization using full-length antisense FGF-23 cRNA as a probe, we showed that abundant amounts of FGF-23 protein and mRNA are present in certain tumor cells of five different OOM tumors. CONCLUSIONS: We conclude that OOM tumors express FGF-23 protein and that the immunohistochemical detection of FGF-23 in OOM tumors is feasible and may help in establishing the diagnosis of tumor-induced hypophosphatemia through analysis of biopsies or surgical specimens. PMID- 12590647 TI - Cell-specific expression of NADPH-dependent cytosolic 3,5,3'-triiodo-L-thyronine binding protein (p38CTBP). AB - We have previously shown that cytosolic 3,5,3'-triiodo-L-thyronine (T3)-binding protein (CTBP) possesses a high affinity for T3 binding in the presence of nicotinamide adenine dinucleotide phosphate in vitro, and that p38CTBP increases intracellular content of T3, and suppresses T3-mediated transactivity. Screening of mRNA expression in 73 different human tIssues has demonstrated that p38CTBP mRNA is expressed at high levels in brain and heart. We have examined the intracellular localization and tissue-specific distribution of this protein by using a specific antibody against human p38CTBP. Western blotting and immunoprecipitation studies have shown that the antibody recognizes human p38CTBP. Interaction of p38CTBP with the antibody did not affect the T3-binding activity of p38CTBP, and its dimer formation in vitro. Western blotting analysis has shown that p38CTBP is expressed in brain and heart predominantly, similar to the distribution of mRNA. Immunohistochemical studies have demonstrated p38CTBP in neural cells and cardiac muscle cells. p38CTBP localizes in cytoplasm rather than in nuclei in neural cells. The evidence for the presence of tIssue-specific localization of p38CTBP has indicated that p38CTBP has a tIssue-specific function, such as the regulation of T3 delivery from cytoplasm to nuclei. PMID- 12590649 TI - Steroid receptor coactivator-3 glutamine repeat polymorphism and the androgen insensitivity syndrome. PMID- 12590650 TI - Purification and identification of a binding protein for pancreatic secretory trypsin inhibitor: a novel role of the inhibitor as an anti-granzyme A. AB - Pancreatic secretory trypsin inhibitor (PSTI) is a potent trypsin inhibitor that is mainly found in pancreatic juice. PSTI has been shown to bind specifically to a protein, distinct from trypsin, on the surface of dispersed cells obtained from tissues such as small intestine. In the present study, we affinity-purified the binding protein from the 2% (w/v) Triton X-100-soluble fraction of dispersed rat small-intestinal cells using recombinant rat PSTI. Partial N-terminal sequencing of the purified protein gave a sequence that was identical with the sequence of mouse granzyme A (GzmA), a tryptase produced in cytotoxic lymphocytes. We confirmed the formation of an affinity-cross-linked complex between (125)I labelled PSTI and recombinant rat GzmA (rGzmA). In situ hybridization and immunostaining revealed the existence of GzmA-expressing intraepithelial lymphocytes in the rat small intestine. We concluded that the PSTI-binding protein isolated from the dispersed cells is GzmA that is produced in the lymphocytes of the tissue. The rGzmA hydrolysed the N -alpha-benzyloxycarbonyl-L lysine thiobenzyl ester (BLT), and the BLT hydrolysis was inhibited by PSTI. Sulphated glycosaminoglycans, such as fucoidan or heparin, showed almost no effect on the inhibition of rGzmA by PSTI, whereas they decreased the inhibition by antithrombin III. In the present paper, we propose a novel role of PSTI as a GzmA inhibitor. PMID- 12590651 TI - Rho family GTPase Rnd2 interacts and co-localizes with MgcRacGAP in male germ cells. AB - The male-germ-cell Rac GTPase-activating protein gene (MgcRacGAP) was initially described as a human RhoGAP gene highly expressed in male germ cells at spermatocyte stage, but exhibits significant levels of expression in most cell types. In somatic cells, MgcRacGAP protein was found to both concentrate in the midzone/midbody and be required for cytokinesis. As a RhoGAP, MgcRacGAP has been proposed to down-regulate RhoA, which is localized to the cleavage furrow and midbody during cytokinesis. Due to embryonic lethality in MgcRacGAP -null mutant mice and to the lack of an in vitro model of spermatogenesis, nothing is known regarding the role and mode of action of MgcRacGAP in male germ cells. We have analysed the expression, subcellular localization and molecular interactions of MgcRacGAP in male germ cells. Whereas MgcRacGAP was found only in spermatocytes and early spermatids, the widespread RhoGTPases RhoA, Rac1 and Cdc42 (which are, to various extents, in vitro substrates for MgcRacGAP activity) were, surprisingly, not detected at these stages. In contrast, Rnd2, a Rho family GTPase-deficient G-protein was found to be co-expressed with MgcRacGAP in spermatocytes and spermatids. MgcRacGAP was detected in the midzone of meiotic cells, but also, unexpectedly, in the Golgi-derived pro-acrosomal vesicle, co localizing with Rnd2. In addition, a stable Rnd2-MgcRacGAP molecular complex could be evidenced by glutathione S-transferase pull-down and co immunoprecipitation experiments. We conclude that Rnd2 is a probable physiological partner of MgcRacGAP in male germ cells and we propose that MgcRacGAP, and, quite possibly, other RhoGAPs, may participate in signalling pathways involving Rnd family proteins. PMID- 12590652 TI - Intravenous immunoglobulin in the treatment of primary trigeminal neuralgia refractory to carbamazepine: a study protocol [ISRCTN33042138]. AB - BACKGROUND: We have recently reported successful treatment of patients with chronic pain syndromes using human pooled intravenous immunoglobulin (IVIG) in a prospective, open-label cohort study. A randomised, placebo controlled, double blinded study is needed to confirm these results. We chose to study patients with carbamazepine resistant primary Trigeminal Neuralgia (rpTN), as these had responded particularly well to IVIG.A protocol involving the use of IVIG in rpTN is complex for three reasons: 1. The effect of IVIG does not follow simple dose response rules; 2. The response pattern of patients to IVIG was variable and ranged between no effect at all and pain free remission between two weeks and >1 year; 3. TN is characterized by extremely severe pain, for which operative intervention is (if temporarily) helpful in most patients. DESIGN: A placebo controlled, parallel, add-on model was developed and the primary outcome variable defined as the length of time during which patients remain in the study. Study groups are compared using Kaplan-Maier survival analysis. Patients record their response to treatment ("severe, moderate, slight, no pain"). The study coordinator monitors pain diaries. Severe or moderate pain of three days duration will result in termination of the study for that patient. CONCLUSIONS: This study design utilizes a method of survival analysis and is novel in chronic pain research. It allows for both early departure from the study and voluntary crossover upon non-response. It may be applicable to the analysis of IVIG efficacy in other chronic pain syndromes. PMID- 12590653 TI - Management of congestive heart failure: a gender gap may still exist. Observations from a contemporary cohort. AB - BACKGROUND: Unlike other cardiovascular diseases the incidence and prevalence of congestive heart failure (CHF) continues to increase. While gender differences in coronary artery disease have been well described, to date, there has been a relative paucity of similar data in patients with CHF. We conducted a pilot study to evaluate the profile and management of patients with CHF at a tertiary care centre to determine if a gender difference exists. METHODS: A chart review was performed at a tertiary care centre on consecutive patients admitted with a primary diagnosis of CHF between June 1997 and 1998. Co-morbidity, diagnostic investigations, and management of CHF were recorded. Comparisons between male and female patients were conducted. RESULTS: One hundred and forty five patients were reviewed. There were 80 male (M) and 65 female (F) patients of similar age [71.6 vs. 71.3 (M vs. F), p = NS]. Male patients were more likely to have had a previous myocardial infarction (66% vs. 35%, p < 0.01) and revascularization (41% vs. 20%, p < 0.05), and had worse left ventricular ejection fraction (LVEF) than women, [median LVEF 3 vs. 2 (M vs. F), p < 0.01]. Male patients were more likely to have a non-invasive assessment of left ventricular (LV) function [85% vs. 69%, (M vs. F), p < 0.05]. A logistic regression analysis suggests that amongst those without coronary disease, males were more likely to receive non-invasive testing. There were no differences in the use of prescribed medications, in this cohort. CONCLUSIONS: This pilot study demonstrated that there seem to be important gender differences in the profile and management of patients with CHF. Importantly women were less likely to have an evaluation of LV function. As assessment of LV function has significant implications on patient management, this data justifies the need for larger studies to assess gender differences in CHF profile and treatment. PMID- 12590654 TI - Ancient conserved domains shared by animal soluble guanylyl cyclases and bacterial signaling proteins. AB - BACKGROUND: Soluble guanylyl cyclases (SGCs) are dimeric enzymes that transduce signals downstream of nitric oxide (NO) in animals. They sense NO by means of a heme moiety that is bound to their N-terminal extensions. RESULTS: Using sequence profile searches we show that the N-terminal extensions of the SGCs contain two globular domains. The first of these, the HNOB (Heme NO Binding) domain, is a predominantly alpha-helical domain and binds heme via a covalent linkage to histidine. Versions lacking this conserved histidine and are likely to interact with heme non-covalently. We detected HNOB domains in several bacterial lineages, where they occur fused to methyl accepting domains of chemotaxis receptors or as standalone proteins. The standalone forms are encoded by predicted operons that also contain genes for two component signaling systems and GGDEF-type nucleotide cyclases. The second domain, the HNOB associated (HNOBA) domain occurs between the HNOB and the cyclase domains in the animal SGCs. The HNOBA domain is also detected in bacteria and is always encoded by a gene, which occurs in the neighborhood of a gene for a HNOB domain. CONCLUSION: The HNOB domain is predicted to function as a heme-dependent sensor for gaseous ligands, and transduce diverse downstream signals, in both bacteria and animals. The HNOBA domain functionally interacts with the HNOB domain, and possibly binds a ligand, either in cooperation, or independently of the latter domain. Phyletic profiles and phylogenetic analysis suggest that the HNOB and HNOBA domains were acquired by the animal lineage via lateral transfer from a bacterial source. PMID- 12590656 TI - Larval stages of Brachylaima fuscatum in the terrestrial snail Limicolaria aurora from southern Nigeria. AB - Of 150 specimens of the gastropod snail Limicolaria aurora examined from the Edo and Delta states of Nigeria, 63.4% were infected with larval digeneans comprising mother sporocysts (12.1%) daughter sporocysts (20.4%) cercariae (43.1%) and metacercariae (24.5%). Attempts to experimentally infect three 14-day-old chicks (Gallus domesticus) and two laboratory-bred 4-month-old mice (Mus musculus) by oral feeding and peritoneal injection with cercariae were negative, although experimental infections of chicks via a cloacal drop yielded 62 immature and 37 mature worms from the intestinal caeca and ileum. The worms were identified as Brachylaima fuscatum (Trematoda: Brachylaimidae). The study also revealed that L. aurora acts as an intermediate host for B. fuscatum, in addition to Eulota sp., Helix sp., Helicella sp., Oxychilus sp. and Agrolimax sp. PMID- 12590657 TI - Population dynamics of Lymnaea columella and its natural infection by Fasciola hepatica in the State of Minas Gerais, Brazil. AB - Studies on the population dynamics and natural infection of Lymnaea columella by Fasciola hepatica were carried out from September 1999 to December 2000 in a low lying area near Itajuba in the Brazilian State of Minas Gerais. A total of 626 snails were collected monthly at nine different sites, counted, and dissected to search for larvae of F. hepatica. The highest populations of L. columella were reached in October of 1999 and August of 2000, and the highest natural infection rates of snails by F. hepatica were reached in September 1999 (5.2%) and July 2000 (3.9%). The removal by farmers of aquatic plants from the drainage furrows caused a drastic reduction in this snail population. PMID- 12590655 TI - Compensatory mutations cause excess of antagonistic epistasis in RNA secondary structure folding. AB - BACKGROUND: The rate at which fitness declines as an organism's genome accumulates random mutations is an important variable in several evolutionary theories. At an intuitive level, it might seem natural that random mutations should tend to interact synergistically, such that the rate of mean fitness decline accelerates as the number of random mutations is increased. However, in a number of recent studies, a prevalence of antagonistic epistasis (the tendency of multiple mutations to have a mitigating rather than reinforcing effect) has been observed. RESULTS: We studied in silico the net amount and form of epistatic interactions in RNA secondary structure folding by measuring the fraction of neutral mutants as a function of mutational distance d. We found a clear prevalence of antagonistic epistasis in RNA secondary structure folding. By relating the fraction of neutral mutants at distance d to the average neutrality at distance d, we showed that this prevalence derives from the existence of many compensatory mutations at larger mutational distances. CONCLUSIONS: Our findings imply that the average direction of epistasis in simple fitness landscapes is directly related to the density with which fitness peaks are distributed in these landscapes. PMID- 12590658 TI - Fasciola gigantica: larval productivity of three different miracidial isolates in the snail Lymnaea truncatula. AB - Bimiracidial infections of Lymnaea truncatula with three isolates of Fasciola gigantica, originating from China, Egypt and Madagascar, were carried out to determine the effect of geographic origin of the parasite on the larval productivity of redial generations. The prevalences of experimental infections in snails exposed to strains from Madagascar, China and Egypt were 20.8%, 60.0% and 80.0%, respectively. At day 49 post-exposure (p.e.), the total number of free rediae in snails infected with the Egyptian isolate was significantly higher than that recorded in the Madagascan group. On the other hand, at day 49 p.e., the majority of cercariae in the Chinese and Egyptian groups were produced by R2a rediae (70.6% and 66.6% of cercariae produced by all live rediae), while, in the Madagascan group, the cercariae were produced mainly by the first redial generation. Snails infected with the Egyptian isolate of miracidia developed more live rediae and, consequently, could produce a higher number of cercariae. As a result, L. truncatula snails were highly adapted to infections with the Egyptian and Chinese isolates of F. gigantica. PMID- 12590659 TI - Distribution of intestinal parasitoses in relation to environmental and sociocultural parameters in La Plata, Argentina. AB - Environmental parameters influencing the distribution of parasite species in three neighbourhoods of differing socioeconomic conditions in La Plata, Argentina were analysed. Coproparasitological screenings were performed in children up to 14 years old from a marginal zone (100), a suburban neighbourhood (101), and an urban area (91) in 1999--2000. The presence of parasite species in environmental samples (water and soil) and the degree of association among parasite communities was documented and evaluated. The prevalence of infection in each population was 73.0%, 54.4% and 35.2%, respectively. The frequencies of helminths and pathogenic protozoa were both higher in the marginal zone, where sanitary and environmental conditions were significantly inferior compared with the other zones. The high prevalence of intestinal parasites in this infantile population was related to parasitic contamination of the soil and water sources in addition to deficient sanitary and sociocultural conditions. Calculation of an equitability index revealed that the specific richness was less equitable once socioeconomic conditions and hygienic practices were improved. This study demonstrates the need to implement management practices for the control of intestinal parasitoses in accordance with the environmental and sociocultural characteristics of a given ecosystem. PMID- 12590661 TI - Penetration of human skin by the cercariae of Schistosoma mansoni: an investigation of the effect of multiple cercarial applications. AB - It has previously been postulated that L-arginine emitted by penetrating Schistosoma mansoni cercariae serves as an intraspecific signal guiding other cercariae to the penetration site. It was suggested that penetrating in groups offers a selective advantage. If this hypothesis is correct and group penetration at one site on the host offers an advantage, it would follow that at such a site, successive groups of cercariae would be able to penetrate skin in either greater numbers or at a faster rate. This prediction was tested by the use of an in vitro model of cercarial penetration based on the Franz cell and using human skin. It was demonstrated that there was no increase in the percentage of cercariae able to penetrate the skin with subsequent exposures. Consequently, it seems unlikely that the release of L-arginine by cercariae during penetration could have evolved as a specific orientation system based on a selective advantage offered by group penetration. PMID- 12590660 TI - Involvement of cysteine proteinases in excystment of Paragonimus ohirai metacercariae induced by sodium cholate and A23187. AB - The involvement of intrinsic proteinases in the excystment of Paragonimus ohirai metacercariae was studied in in vitro excystment induced by sodium (Na) cholate, a bile salt and A23187, a Ca2+ ionophore. The effects of various proteinase inhibitors on the in vitro excystment were examined and similar inhibitory profiles were obtained. Benzyloxycarbonyl-L-leucyl-L-leucinal (Z-Leu-Leu-H), a cysteine proteinase inhibitor and 4-(2-aminoethyl)-benzenesulfonyl fluoride (Pefabloc SC), a serine proteinase inhibitor completely inhibited excystment, while L-3-carboxy-2,3-trans-epoxypropionyl-leucylamido (4-guanidino)-butane (E 64), a cysteine proteinase inhibitor and leupeptin, a cysteine/serine proteinase inhibitor permitted partial excystment at a lower rate, but inhibited it from proceeding from the partial excystment stage. In secretions released from metacercariae during excystment, proteinase activities detected towards various fluorogenic peptidyl substrates were almost completely inhibited by Z-Leu-Leu-H and E-64, but not by Pefabloc SC. Sodium cholate induced a higher secretion of cysteine proteinases and a higher rate of excystment than A23187. Profiles of cysteine proteinase activities towards five peptidyl substrates detected were markedly different among the two secretions and the lysate of newly excysted juveniles. Newly excysted juveniles released cysteine proteinases with similar activity profiles and levels to metacercariae induced by Na cholate-incubation, whereas the release of cysteine proteinases was reduced compared with metacercariae induced by A23187-incubation. These results provide valuable information about the involvement of intrinsic proteinases in metacercarial excystment. PMID- 12590662 TI - A molecular phylogeographic study based on DNA sequences from individual metacercariae of Paragonimus mexicanus from Guatemala and Ecuador. AB - A molecular phylogeographic study of Paragonimus mexicanus collected from Guatemala and Ecuador was performed. Genomic DNA was extracted from individual metacercariae, and two gene regions (partial mitochondrial cytochrome c oxidase subunit 1 (CO1) and the second internal transcribed spacer of the nuclear ribosomal gene repeat (ITS2)) were amplified by the polymerase chain reaction (PCR). Sequences segregated in a phylogenetic tree according to their geographic origins. ITS2 sequences from Ecuador and Guatemala differed at only one site. Pairwise distances among CO1 sequences within a country were always lower than between countries. Nevertheless, genetic distances between countries were less than between geographical forms of P. westermani that have been suggested to be distinct species. This result suggests that populations from Guatemala and Ecuador are genetically differentiated perhaps at the level of subspecies. PMID- 12590663 TI - Taenia solium taeniasis/cysticercosis in Papua, Indonesia in 2001: detection of human worm carriers. AB - A preliminary study to detect human worm carriers of Taenia solium in Papua (Irian Jaya), Indonesia was carried out using stool examinations for the detection of copro-antigens and adult proglottids after chemotherapy, and confirmation by mitochondrial DNA analysis using expelled proglottids and metacestodes developed in NOD/Shi-scid mice from eggs of expelled proglottids. Approximately 8.6% of the local population in Kama (5/58), 1 km from the local capital city centre, Wamena, were confirmed to harbour adult T. solium using these techniques. PMID- 12590664 TI - In vitro metabolism of an insect neuropeptide by homogenates of the nematode Caenorhabditis elegans. AB - The cytosolic fraction of homogenates from the free-living soil nematode Caenorhabditis elegans is capable of metabolizing the insect neuropeptide adipokinetic hormone, a decapeptide blocked at the N-terminus by a pGlu residue. Analysis of digests by RP-HPLC and LC-MS revealed that an initial endoproteolytic cleavage step produced a heptapeptide with an unblocked N-terminus that can serve as a substrate for aminopeptidases. The aminopeptidase activity is depressed in the presence of the inhibitor amastatin; the initial product of the endoproteolytic step accumulates during incubation, and expected aminopeptidase product peptides are reduced in amount, as assessed by chromatographic peak size. The absence of some expected peptide fragments in the reaction mixtures suggests that multiple proteases contribute to short peptide half-lives. Comparison of the adipokinetic hormone digestion in C. elegans to that reported previously for insects reveals the same general pattern of peptide fragment production. PMID- 12590665 TI - Cadmium toxicity and snail-digenean interactions in a population of Lymnaea spp. AB - The toxicity of cadmium to a population of Lymnaea peregra and L. stagnalis naturally infected with a range of digeneans and collected from a number of sites in the lower Thames Valley, UK was investigated. Lymnaeid snails were exposed to 100 microg l-1 cadmium and the effects on host survival and emergence of cercariae recorded. Overwintered L. peregra, but not L. stagnalis, showed significantly reduced survival compared to seasonally infected snails, i.e. snails which have acquired an infection during the spring or summer. A significant increase in survival with increasing snail size was demonstrated for L. stagnalis and for seasonally infected L. peregra only. Only L. stagnalis infected with Diplostomum spathaceum and L. peregra infected individually with D. spathaceum, Sanguinicola inermis, Echinoparyphium recurvatum and Notocotylus attenuatus demonstrated a significantly reduced survival compared to laboratory bred controls. The exposure of L. stagnalis to cadmium resulted in a significant reduction in the emergence of D. spathaceum over a 5-day period but cadmium exposed L. peregra showed no difference in the emergence of E. recurvatum cercariae over a 3-day exposure period. The mechanisms and importance of metal toxicity to snail-digenean interactions are discussed. PMID- 12590666 TI - Molecular cloning and expression of the full-length tropomyosin gene from Trichinella spiralis. AB - A clone, designated as TsTM, was selected from the cDNA library of newborn larvae (NBL) of Trichinella spiralis through immunoscreening against infected sera. The clone contained a cDNA transcript of 855 bp in length with a single open reading frame, which encoded 285-amino acids (33 kDa in the estimated molecular weight). A sequence analysis revealed that the clone TsTM encoded the full-length of tropomyosin gene. The phylogenetic analysis of the tropomyosin gene was in good agreement with the classical taxonomical position of T. spiralis. The fusion proteins encoded by the clone TsTM were produced in an Escherichia coli expression system and affinity purified, and the antibody was raised against the protein for the following studies. The antibody against the fusion protein positively bound to the hypodermal muscle layer in immunolocalization analysis, and the 35 kDa band in crude extracts of muscle larvae but not in excretory and secretory (ES) products on Western blots. The antigenicity of the clone TsTM was recognized by host mice but exhibited little species specificity. PMID- 12590667 TI - Increased susceptibility of Atlantic salmon Salmo salar to infections with Gyrodactylus derjavini induced by dexamethasone bath treatment. AB - Dexamethasone, a known immunosuppressant, was administered by bath or injection to Atlantic salmon Salmo salar (Conon stock) to study if this treatment could affect the susceptibility of fish to infections with a Danish strain of Gyrodactylus derjavini (Monogenea). Three groups of S. salar (Conon stock) were immersion treated either with 10, 60 or 240 microg dexamethasone l-1 water, respectively. In addition, one group (positive control) was treated intraperitoneally with 200 microg dexamethasone per fish and one negative control group was kept untreated. A single G. derjavini parasite was placed on the anal fin of each fish and the infection was subsequently monitored weekly for 6 weeks. An increase in parasite populations on the salmon was positively correlated with the amount of immunosuppressant used. Infection levels in the group immersion treated with dexamethasone (240 microg l-1 water) and in the i.p. treated positive control group were significantly higher compared to the untreated control group. PMID- 12590668 TI - Pseudobacciger harengulae from the Atlantic herring Clupea harengus: a new host and locality record. AB - During the summer and autumn of 1994, 1995 and 1996, 406 juvenile herring caught off the Swedish west coast were examined for parasites. Amongst those found was the digenean Pseudobacciger harengulae, which represents new host and locality records for this parasite. Pseudobacciger harengulae has been reported from several species of clupeiformes, mostly from tropical and temperate regions of the Atlantic, Pacific and Indian Oceans. The morphology of P. harengulae is described and compared with earlier descriptions of P. harengulae and P. manteri. The possible relationships between P. harengulae and P. manteri are discussed and the validity of the P. manteri is questioned. Most of the specimens (75%) of P. harengulae were found in the pyloric caeca and the remainder (25%) in the intestine. PMID- 12590669 TI - Setaria cervi: in vitro released collagenases and their inhibition by Wuchereria bancrofti infected sera. AB - In vitro released products of adult Setaria cervi females, microfilariae and extracts showed considerable amounts of collagenase activity. On the basis of per mg protein released in vitro, the products of both microfilariae and adult females exhibited comparable activity but this was much higher than that of extract of microfilariae and adult females. Two collagenase enzymes with molecular masses of 50 kDa and 70 kDa were separated using DEAE-sepharose CL6B and Sephadex G-100 column chromatography. The 50 kDa and 70 kDa collagenase exhibited pH optima of 5.2 and 7.0, respectively. Considering specific activity, the 50 kDa enzyme was found to contribute about ten times more collagenase activity as compared to the 70 kDa enzyme. An inhibition study revealed obvious differences between them. Thiol group inhibitors such as N-ethylmaleimide and leupeptin inhibited the 50 kDa enzyme but this was strongly activated by dithiothreitol, a thiol group stabilizer. Alternatively, the 70 kDa enzyme showed a sensitivity to a metal chelator and a serine group inhibitor indicating its metalloserine protease nature. The antifilarial drug diethylcarbamazine did not demonstrate any inhibition under in vitro conditions. Both enzymes were significantly inhibited by antibody IgG separated from Wuchereria bancrofti infected human sera, showing a possible immunoprotective role. PMID- 12590670 TI - Prevalence of canine dirofilariasis in Taiwan. AB - Between 1993 and 1997, 837 stray dogs from North Taiwan were necropsied and examined for dog heartworm infections. A thick smear from 20 ml of peripheral blood from each dog was also prepared and examined for microfilariae (mf). The overall prevalence of adult worms in the dog population was 57%. The prevalence of mf in 1228 house dogs from different parts of Taiwan was also determined from 20 ml of peripheral blood in the same way. The overall prevalence of mf was 25%, with a value of 30% in the main island of Taiwan, this being 15 times higher than that in the offshore islands (2%). In Taiwan, the prevalence ranged from 4% in Hualien County, East Taiwan, to 41% in Nantou County, Central Taiwan. The mf prevalences on offshore islands were 1% on Liuchiu and 2% on Lanyu. The mf density per 20 ml blood in 82 house dogs was found to be 23 mf per dog, with a range of 3-97 mf per dog. A total of 477 stray dogs were found to be infected with adult worms of Dirofilaria immitis. The mean number of 7 worms per dog was obtained, with a range of 1-55 worms per dog. These results indicate that the prevalence of canine dirofilariasis has increased in Taiwan over the past 10 years. Moreover, the prevalence may be related to the wind speed, temperature, relative humidity, and altitude in the different areas surveyed. PMID- 12590671 TI - On the longevity of Schistosoma curassoni. AB - It is demonstrated that Schistosoma curassoni, a parasite of sheep, cattle and goats in parts of West Africa, will live for at least 8 years 5 months in a sheep. The sheep was exposed to 500 cercariae of S. curassoni liberated from infected Bulinus wrighti. The sheep died of natural causes, and at post-mortem 28 pairs of adult S. curassoni were removed from the mesenteric and rectal veins. All female worms were gravid, and eggs were hatched from faeces to produce miracidia. The development of immune responses of the host had apparently little or no effect on the viability of the eggs. Histological studies of the liver, small and large intestines revealed mild pathological symptoms. The longevity of S. curassoni is the first record of longevity of schistosomes to be based on worm counts. PMID- 12590672 TI - Virtual reality surgery: neurosurgery and the contemporary landscape. AB - OBJECTIVE: Virtual reality-simulated environments have been used for the training of personnel, most notably for military applications, for more than 35 years. The advantages conferred by being able to train novice personnel in a low- to no-risk simulated environment have long been appreciated by the medical community. The recent availability of affordable gigahertz-range microprocessors (once the exclusive domain of the Cray supercomputer) has made photorealistic graphical rendering and manipulation of virtual surgical substrates a reality. Concomitant advances in artificial intelligence systems and the portability of patient specific magnetic resonance imaging, computed tomographic scanning, and angiographic image data presage the emergence of the surgical simulator as a modern surgical training adjunct. An overview of the status of surgical simulation with regard to its adaptability to current surgical training regimens is presented. METHODS: Extensive MEDLINE, Internet, and other database searches spanning the years 1960 to 2002 were conducted in an effort to delineate the status of simulated surgical environments. RESULTS: As would be expected, most articles addressing surgical simulation as their primary focus have been published in the past decade. A review of this literature demonstrates the broadest application in the field of endoscopic (and laparoscopic) procedures, most likely as a result of the reduced engineering burden with respect to incorporation of a haptic interface. CONCLUSION: The realization of ergonomically acceptable haptic interfaces remains elusive. Improvements in graphical rendering and the incorporation of artificial intelligence functions signal the certain emergence of surgical simulators as a viable supplement to the Halstedian method of surgical training. PMID- 12590673 TI - A three-dimensional interactive virtual dissection model to simulate transpetrous surgical avenues. AB - OBJECTIVE: This project involves the development of a three-dimensional surgical simulator called interactive virtual dissection, which is designed to teach surgeons the visuospatial skills required to navigate through a transpetrosal approach. METHODS: A robotically controlled microscope is used for surgical planning and data collection. The spatial anatomic data are recorded from sequentially deeper cadaveric head dissections as a series of superimposed anatomic pictures in stereoscopic digital format. The sequential series of images are then merged to form the final virtual representation. RESULTS: The current three-dimensional virtual reality simulator allows the user to drill the petrous bone progressively deeper and to identify crucial structures much like an experienced surgeon drilling the petrous bone. The program allows surgeons and trainees to manipulate the virtual "surgical field" by interacting with the surgical anatomy. The interactive system functions on a desktop computer. CONCLUSION: The ability to visualize and understand anatomic spatial relationships is crucial in surgical planning, as is a surgeon's confidence in performing the surgery. The virtual reality simulator does not replace the need for practicing surgery on cadavers. However, it is designed to facilitate, via stereoscopic projection, learning how to manipulate a drill in complicated or unfamiliar surgical approaches (e.g., a transpetrosal approach). PMID- 12590674 TI - Incidence and clinical evolution of postoperative deficits after volumetric stereotactic resection of glial neoplasms involving the supplementary motor area. AB - OBJECTIVE: We report the incidence and clinical evolution of postoperative deficits and supplementary motor area (SMA) syndrome after volumetric stereotactic resection of glial neoplasms involving the posterior one-third of the superior frontal convolution. We investigated variables that may be associated with the occurrence of SMA syndrome. METHODS: The postoperative clinical status of 27 consecutive patients who underwent resection of SMA gliomas was retrospectively reviewed. Neurological examination results were recorded 1 day, 1 week, 1 month, and 6 months postoperatively. The extent of tumor resection, the percentage of SMA resection, violation of the cingulate gyrus, and operative complications were tabulated. RESULTS: The overall incidence of SMA related deficits was 26% (7 of 27 patients), with 3 patients having complete SMA syndrome and 4 patients having partial SMA syndrome. Two additional patients (7.5%) had other postoperative deficits, including one with mild facial weakness and one with transient aphasia. The resection of low-grade gliomas was associated with a higher incidence of SMA syndrome, an outcome that likely reflects more complete removal of functional SMA cortex in this subset of patients. Intraoperative monitoring localized the precentral sulcus within the preoperatively defined tumor volume in 6 (22%) of 27 patients, thereby precluding gross total resection. All 27 patients had excellent outcomes at the 6-month follow-up examination. CONCLUSION: When the resection of SMA gliomas is limited to the radiographic tumor boundaries, the incidence and severity of SMA syndrome may be minimized. With the use of these resection parameters, patients with high grade SMA gliomas are unlikely to experience SMA syndrome. These findings are helpful in the preoperative counseling of patients who are to undergo cytoreductive resection of SMA gliomas. PMID- 12590675 TI - Gamma knife radiosurgery for the treatment of cavernous sinus meningiomas. AB - OBJECTIVE: We report on the efficacy of gamma knife radiosurgery for cavernous sinus meningiomas. METHODS: Between January 1994 and December 1999, we used gamma knife radiosurgery for the treatment of 43 patients with cavernous sinus meningiomas. Forty-two patients were followed up for a mean of 49.4 months (range, 18-84 mo). The patients' average age was 55 years (range, 18-81 yr). Twenty-two patients (52%) underwent operations before radiosurgery, and 20 patients (48%) underwent radiosurgery after the diagnosis was made by magnetic resonance imaging. The tumor volumes ranged from 1.2 to 101.5 cm(3) (mean, 14.7 cm(3)). The tumors either compressed or were attached to the optic apparatus in 17 patients (40.5%). The marginal radiation dose was 8 to 15 Gy (mean, 11 Gy), and the optic apparatus was irradiated with 2 to 12 Gy (mean, 6.2 Gy). Three patients with a mean tumor diameter greater than 4 cm were treated by two-stage radiosurgery. RESULTS: Thirty-eight patients (90.5%) demonstrated tumor growth control during the follow-up period after radiosurgery. Tumor regression was observed in 25 patients (59.5%), and growth was unchanged in 13 patients (31%). Regrowth or recurrence occurred in four patients (9.5%). The actual tumor growth control rate at 5 years was 92%. Only one patient (2.4%) experienced regrowth within the treatment field; in other patients, regrowth occurred at sites peripheral to or outside the treatment field. Twelve patients (28.6%) had improved clinically by the time of the follow-up examination. None of the patients experienced optic neuropathy caused by radiation injury or any new neurological deficits after radiosurgery. CONCLUSION: Gamma knife radiosurgery may be a useful option for the treatment of cavernous sinus meningiomas not only as an adjuvant to surgery but also as an alternative to surgical removal. We have shown it to be safe and effective even in tumors that adhere to or are in close proximity to the optic apparatus. PMID- 12590676 TI - Neuroendoscopic treatment for colloid cysts of the third ventricle: the experience of a decade. AB - OBJECTIVE: Microsurgical resection or ventriculoperitoneal shunt placement was for a long time the only means of treatment for patients with colloid cysts. In the past few years, however, endoscopic procedures have gained increasing significance and have been used more widely. Long-term results are now available for the first time, which enabled us to evaluate this method and assess its future relevance. METHODS: Twenty patients with symptomatic colloid cysts of the third ventricle have been treated endoscopically in our department during the past 10 years. Retrospective analysis and follow-up of the patients' clinical and radiological outcomes were performed. RESULTS: In the early postoperative period, 18 patients had excellent outcomes, with clinical signs improving immediately. One patient experienced intraoperative hemorrhage followed by temporary postoperative psychosis and IIIrd cranial nerve palsy. Another patient remained shunt-dependent because of aseptic meningitis after the endoscopic procedure. In the long-term follow-up, one patient had to be reoperated to treat cyst recurrence. The operative time was strongly dependent on the cyst material as well as on the surgeon's experience with endoscopic techniques; it varied between 60 and 300 minutes (mean operative time, 200 min). The average hospitalization time was 9 days. Long-term follow-up ranging from 1 to 10 years showed a clear benefit in each patient. In three individuals with pre- and postoperative short term memory deficits and in one patient who complained of headaches, symptoms resolved gradually during the first few months after surgery. The patient with intraoperative hemorrhage complained of slight permanent short-time memory deficit. The analysis of postoperative computed tomographic and magnetic resonance imaging scans revealed a remaining cyst wall in the majority of patients and an inconsistent decrease in ventricular size. Postoperative cine magnetic resonance imaging studies showed normalization of cerebrospinal fluid flow in 17 patients. CONCLUSION: Continued improvement of endoscopic techniques and instruments, together with good long-term results in endoscopically treated patients, have established this method as an alternative to microsurgical techniques and might even set a new standard for treatment. PMID- 12590677 TI - Extensive experience with dural sinus thrombosis. AB - OBJECTIVE: Dural sinus thrombosis (DST) is an uncommon cause of stroke. The safest and most effective therapy for DST has not been conclusively identified. METHODS: A retrospective chart review of data for 31 patients who were treated for DST at our institution between 1992 and 2001 was performed. Four treatment strategies were identified, i.e., 1). medical observation only, 2). systemic anticoagulation (AC) therapy with heparin, 3). endovascular chemical thrombolysis with urokinase or tissue plasminogen activator and concurrent systemic AC therapy, and 4). mechanical endovascular clot thrombolysis with concurrent systemic AC therapy. Complications and clinical outcomes were assessed for each group. RESULTS: Patients treated solely with medical observation fared the worst; four of five patients experienced intracranial hemorrhagic complications, and only two of five exhibited clinical improvement. Patients who received systemic AC therapy experienced no hemorrhagic complications, even when pretreatment hemorrhage was present; 75% (six of eight patients) exhibited improvement with AC therapy alone. Chemical thrombolysis was very effective in restoring sinus patency (90% of patients); however, 30% of patients (3 of 10 patients) experienced hemorrhagic complications. Sixty percent of patients (6 of 10 patients) who underwent chemical thrombolysis exhibited clinical improvement. Patients who underwent mechanical thrombectomies demonstrated a low hemorrhagic complication rate, and most (88%) made good recoveries. CONCLUSION: Therapy directed at the underlying clot in DST must begin without delay. Our results suggest that supportive medical management of DST, without therapy directed at the clot or clotting process, is not effective. Systemic AC therapy, even in the presence of intracerebral hemorrhage, seems to be safe. Heparin can be safely titrated to yield partial thromboplastin times of 60 to 70 seconds. Chemical clot thrombolysis is efficacious in opening occluded sinuses but may cause intracranial hemorrhage. We currently recommend either systemic AC therapy or systemic AC therapy in conjunction with mechanical clot thrombectomy as a safe effective treatment for DST. PMID- 12590678 TI - Seizures after aneurysmal subarachnoid hemorrhage treated with coil embolization. AB - OBJECTIVE: We sought to determine the incidence of seizures among patients treated with endovascular coil embolization for ruptured intracranial aneurysms because data on which to base antiepileptic drug (AED) prescriptions and advice to patients regarding driving motor vehicles and other high-risk activities are currently lacking. METHODS: We conducted a single-institute, single-operator observational study of 243 patients referred for endovascular treatment after aneurysmal subarachnoid hemorrhage. Prospective data collection was performed, and all successfully treated patients were followed. The incidence of seizures was compared with published surgical data, and logistic regression analysis of potential clinical associations was performed. Patients were followed for up to 7.7 years (mean follow-up period, 21.9 mo). RESULTS: Ictal seizures occurred at the time of subarachnoid hemorrhage in 26 (11%) of 243 patients and correlated with middle cerebral artery aneurysm location, loss of consciousness at ictus, and AED prescription. No patients experienced periprocedural seizures during their hospitalization. Seven of 233 successfully treated patients (3%) experienced seizures more than 30 days after treatment: late seizures occurred de novo in four patients (1.7%) and in three patients (1.4%) were caused by preexisting epilepsy. Two patients (0.85%) who had de novo seizures developed epilepsy. Late seizures correlated with a history of previous seizures, the presence of a cerebrospinal fluid shunt, and the use of AEDs. CONCLUSION: The low incidence of seizures does not justify the use of prophylactic AED therapy after aneurysmal subarachnoid hemorrhage in patients treated solely with coil embolization, nor does it justify subsequent restrictions on the driving of motor vehicles if the patient is otherwise fit to drive. PMID- 12590679 TI - Small unruptured cerebral aneurysms presenting with oculomotor nerve palsy. AB - OBJECTIVE: Symptomatic unruptured aneurysms have been considered at relatively high risk for future rupture, and the majority of aneurysms that cause symptoms of mass effect are large. Unruptured aneurysms smaller than 1 cm in diameter sometimes cause neurological symptoms, but their clinical aspects remain obscure. In this article, we review our experience with small unruptured aneurysms presenting with oculomotor nerve palsy. METHODS: Sixteen patients with unruptured aneurysms smaller than 1 cm presenting with oculomotor nerve palsy were included in this study. The patients' clinical profiles were reviewed, and factors affecting the recovery of oculomotor function were determined. RESULTS: The mean size of the aneurysms was 5.8 +/- 1.4 mm. Eleven patients (68.8%) had preceding retrobulbar pain. Fifteen patients underwent successful microsurgical clipping or intravascular embolization, but one patient died of aneurysm rupture before surgery. Seven patients (43.8%) had a complete recovery of oculomotor function, six (37.5%) had an incomplete recovery, and two (12.5%) remained unchanged after treatment. The mean interval between the onset of oculomotor nerve palsy and treatment was 4.7 +/- 3.3 days in patients with complete recovery, 24.2 +/- 15.5 days in patients with incomplete recovery, and 41.0 +/- 12.7 days in unchanged patients. Early surgery resulted in more complete recovery of neural function (P < 0.01). CONCLUSION: Unruptured aneurysms smaller than 1 cm can cause neurological symptoms of mass effect. We recommend timely surgery, preferably within 5 days, to avoid not only aneurysm rupture but also functional disability even in patients with unruptured aneurysms smaller than 1 cm. PMID- 12590680 TI - Surgical complications in patients with Parkinson's disease after posteroventral pallidotomy. AB - OBJECTIVE: To investigate the potential operative morbidity in posteroventral pallidotomy (PVP) for patients with Parkinson's disease. METHODS: We designed a retrospective study that included 796 consecutive patients (mean age, 64.9 yr; male, 559; female, 237) with Parkinson's disease. All PVPs (simultaneous bilateral PVP, n = 272; sequential bilateral PVP, n = 88; unilateral PVP, n = 436) were performed during a 7-year period. The total number of operations was 884, and the number of PVP procedures was 1156. In 108 patients, ventral intermediate nucleus thalamotomy was performed simultaneously. RESULTS: The overall complication rate, including temporary problems, was 15.3% of 884 operations. Permanent complications occurred in 3.6% of total operations. Intracranial hemorrhage occurred in 24 operations (2.7%). In seven of them, the patients required craniotomy and hematoma evacuation and sustained a disabling motor deficit (0.8%). Intracranial hemorrhage occurred more often in patients who underwent microelectrode recording and had a history of chronic hypertension. Hemiparesis without intracranial hematoma occurred in 12 operations (1.4%). Microelectrode recording was a risk factor for postoperative hemiparesis without hemorrhage. In 19 operations (2.1%), patients developed a partial visual field deficit. Speech disturbance after surgery was observed in 23 operations (2.6%) but resolved in 17 by 1 week after surgery. In 55 operations (6.2%), patients developed postoperative confusion. This occurred more often in elderly patients and those with advanced disease. In 17 operations (1.9%), patients required observation in the intensive care unit because of postoperative hypotension. CONCLUSION: Complications from stereotactic pallidotomy were not frequent. However, the residual symptoms from complications can be serious in many cases. PMID- 12590681 TI - Automated, patient-interactive, spinal cord stimulator adjustment: a randomized controlled trial. AB - OBJECTIVE: Programmable, multicontact, implanted stimulation devices represent an important advance in spinal cord stimulation for the management of pain. They facilitate the technical goal of covering areas of pain by stimulation-evoked paresthesiae. Adjustment after implantation requires major investments of time and effort, however, if the capabilities of these devices are to be used to full advantage. The objective of maximizing coverage should be met while using practitioners' time efficiently. METHODS: We have developed a patient interactive, computerized system designed for greater ease and safety of operation, compared with the standard external devices used to control and adjust implanted pulse generators. The system automatically and rapidly presents to the patient the contact combinations and pulse parameters specified by the practitioner. The patient adjusts the amplitude of stimulation and then records drawings of stimulation paresthesiae (for comparison with pain drawings), followed by visual analog scale ratings for each setting. Test results are analyzed and sorted to determine the optimal settings. We compared the automated, patient-interactive system with traditional, practitioner-operated, manual programming methods in a randomized controlled trial at two study centers, with 44 patients. RESULTS: The automated, patient-interactive system yielded significantly (P < 0.0001) better technical results than did traditional manual methods, in achieving coverage of pain by stimulation paresthesiae (mean 100 point visual analog scale ratings of 70 and 46, respectively). The visual analog scale ratings were higher for automated testing for 38 patients, higher for manual testing for 0 patients, and equal (tied) for 6 patients. Multivariate analysis demonstrated that the advantage of automated testing occurred independently of practitioner experience; the advantage was significantly greater, however, for experienced patients. The rate of testing (number of settings tested per unit time) was significantly (P < 0.0001) greater for the automated system, in comparison with the rate with a human operator using traditional, manual, programming methods (mean of 0.73 settings/min versus 0.49 settings/min). The automated system also identified settings with improved estimated battery life (and corresponding anticipated cost savings). No complications were observed with automated testing; one complication (transient discomfort attributable to excessive stimulation) occurred with manual testing. CONCLUSION: Automated, patient-interactive adjustment of implanted spinal cord stimulators is significantly more effective and more efficient than traditional manual methods of adjustment. It offers not only improved clinical efficacy but also potential cost savings in extending implanted battery life. It has the additional potential advantages of standardization, quality control, and record keeping, to facilitate clinical research and patient care. It should enhance the clinical application of spinal cord stimulation for the treatment of chronic intractable pain. PMID- 12590682 TI - Risk factors for intracranial hemorrhage among full-term infants: a case-control study. AB - OBJECTIVE: To investigate the cause of intracranial hemorrhage among full-term infants. METHODS: A retrospective, hospital-based, matched case-control study was conducted at London Health Sciences Center, in southwestern Ontario, for the period from January 1, 1985, to December 31, 1996. Cases were diagnosed with magnetic resonance imaging, computed tomography, or ultrasonography within 7 days after birth. Control subjects were matched with respect to year of birth, sex, and, for nontransferred case patients only, obstetrician. RESULTS: Sixty-six full term infants with intracranial hemorrhage were identified, and 104 control subjects were matched. Each factor was independently associated with increased risk of intracranial hemorrhage, as follows: forceps assistance (odds ratio [OR], 4.3; 95% confidence interval [CI], 1.2-15.1), compared with spontaneous vaginal delivery; 1-minute Apgar scores of 1 through 4 (OR, 110; 95% CI, 5.0-2400) and 5 through 8 (OR, 4.9; 95% CI, 1.3-18.3), compared with scores of 9 or 10 (corresponding 5-min Apgar scores were also statistically significant); and requirements for resuscitation (OR, 5.1; 95% CI, 1.8-14.1), compared with no resuscitation requirements. Of the 52 case patients for whom platelet counts were recorded within 48 hours after birth, 30.8% (95% CI, 18.3-43.3%) exhibited counts of less than 70 x 10(9)/L. Platelet counts of less than 50 x 10(9)/L were specifically associated with intraparenchymal hemorrhage and a more severe radiological grade. Forceps-associated hemorrhage was more frequently subarachnoid and subdural and less frequently intraparenchymal. Such hemorrhage also tended to be more caudal in location. CONCLUSION: Thrombocytopenia seems to be an important cause of intraparenchymal hemorrhage, and the use of forceps is more likely to be associated with subarachnoid and subdural hemorrhage. PMID- 12590683 TI - The use of frozen autogenous bone flaps in delayed cranioplasty revisited. AB - OBJECTIVE: To reevaluate the use of frozen autogenous bone flaps for patients undergoing delayed cranioplasty. METHODS: In the past 12 years, 49 patients have undergone delayed cranioplasty using frozen autogenous bone flaps. Bone flaps removed during the initial operation were sealed in three sterilized vinyl bags and stored at -35 degrees C (n = 37) or -84 degrees C (n = 12) for 4 to 168 days (mean, 50.6 d). The bone flaps were thawed at room temperature and replaced in their original positions. After cranioplasty, we monitored resorption of the bone flaps with computed tomography and evaluated the clinical and aesthetic results. Follow-up periods ranged from 14 to 147 months (mean, 59.2 mo). RESULTS: For 47 patients (95.9%), there were no complications during the follow-up period; there was slight thinning of the bone flap in some cases, but clinical and aesthetic results were highly satisfactory. Resorption was observed for a 12-year-old boy who had undergone cranioplasty, using two pieces of bone flap, 66 days after the initial operation. A 14-year-old boy with a cerebral contusion experienced a bone flap infection. Both patients underwent a second cranioplasty procedure, with ceramic plates. CONCLUSION: The clinical and aesthetic results of delayed cranioplasty using frozen autogenous bone flaps were satisfactory. The most important factor for success was excellent contiguity between the flap and the bone edge. PMID- 12590684 TI - Double-insurance bypass for internal carotid artery aneurysm surgery. AB - OBJECTIVE: The aim of this article is to present the usefulness of a double bypass method in the surgical treatment of complex internal carotid artery (ICA) aneurysms. For patients with clippable but complex aneurysms of the ICA having poor collateral circulation, bypass surgery is needed before temporary occlusion of the ICA. We propose a double bypass for safety. METHODS: The superficial temporal artery was anastomosed to the distal cortical branch of the middle cerebral artery (MCA), followed by anastomosis between the radial artery and the inferior trunk of the MCA. For patients with clippable ICA aneurysms, the radial artery was temporarily anastomosed to the inferior trunk of the MCA by raising the ipsilateral forearm to the head after the radial artery was harvested. After the aneurysm had been clipped, the anastomosed radial artery was cut close to the anastomosed site and repositioned back to the original arm. RESULTS: This double bypass procedure was performed in two patients, and no ischemic complications related to revascularization were observed. Temporary occlusion times of the MCA for superficial temporal artery-to-MCA anastomosis and radial artery-to-MCA anastomosis were 30 and 46 minutes in one patient and 28 and 55 minutes in another. CONCLUSION: This surgical procedure, which we called "double-insurance bypass," can reduce the risk of ischemic complications associated with revascularization of the ICA. PMID- 12590685 TI - Intraoperative spinal digital subtraction angiography: technique and results. AB - OBJECTIVE: To evaluate technical and methodological aspects of intraoperative spinal digital subtraction angiography (ISDSA) in our clinical practice and to assess its practicability, safety, and accuracy for the surgical treatment of spinal vascular malformations. METHODS: Between August 1997 and February 2002, a total of 30 patients were treated either surgically (n = 18) or endovascularly (n = 12) for spinal vascular lesions at our institution. The clinical records of five patients who underwent ISDSA were analyzed retrospectively. The thoracic segment was involved in three patients and the medullary cone in two. RESULTS: ISDSA could be performed in four cases. In one patient, the segmental artery could not be probed sufficiently while the patient was prone. No complications occurred from the application of ISDSA. The method was beneficial for the neurosurgeon in all but one patient because the vascular anatomy of the malformation was shown with respect to the surgical approach, including the nidus, and immediate resection control could be performed before wound closure. The duration of the procedure was prolonged by 45 minutes on average. CONCLUSION: ISDSA is safe and effective, especially in surgery for complex vascular and recurrent malformations. Benefits to the patient outweigh the additional expense and prolongation of the surgical procedure. PMID- 12590686 TI - Effect of changing patient position from supine to prone on the accuracy of a Brown-Roberts-Wells stereotactic head frame system. AB - OBJECTIVE: Despite the growing popularity of frameless image-guided surgery systems, stereotactic frame systems are widely accepted by neurosurgeons and are commonly used to perform biopsies, functional procedures, and stereotactic radiosurgery. We investigated the accuracy of the Brown-Roberts-Wells stereotactic frame system when the mechanical load on the frame changes between preoperative imaging and the intervention because of different patient position: supine during imaging, prone during intervention. METHODS: We analyzed computed tomographic images acquired from 14 patients who underwent stereotactic biopsy, deep brain stimulator implantation, or radiosurgery. Two images were acquired for each patient, one with the patient in the supine position and one in the prone position. The prone images were registered to the respective supine images by use of an intensity-based registration algorithm, once using only the frame and once using only the head. The difference between the transformations produced by these two registrations describes the movement of the patient's head with respect to the frame. RESULTS: The maximum frame-based registration error between the supine and prone positions was 2.8 mm; it was more than 2 mm in two patients and more than 1.5 mm in six patients. Anteroposterior translation is the dominant component of the difference transformation for most patients. In general, the magnitude of the movement increased with brain volume, which is an index of head weight. CONCLUSION: To minimize frame-based registration error caused by a change in the mechanical load on the frame, stereotactic procedures should be performed with the patient in the identical position during imaging and intervention. PMID- 12590687 TI - Hydrodynamic properties of extraventricular drainage systems. AB - OBJECTIVE: Extraventricular drains (EVDs) are intended to control intracranial pressure for patients with acute disorders of the cerebrospinal fluid circulation. We tested five commercially available EVDs to assess their fundamental hydrodynamic properties, which determine the quality of this control. METHODS: The five most frequently used drainage systems were tested. The pressure responses to water flow from a computer-controlled infusion pump were studied in the rig constructed in the UK Shunt Evaluation Laboratory (Cambridge, England). EVDs were studied under normal conditions and after brief (20-s) contact of the vent located in the drip chamber with the test reagent. Pure water and water with 10% rat blood content were used for testing. RESULTS: All of the tested EVDs demonstrated low hydrodynamic resistance [<3.5 mm Hg/(ml/min)], indicating their ability to control intracranial pressure. When the drip chamber vents were in brief contact with the reagent, the hydrodynamic properties of two models were unaffected. For the three other EVDs, blockage of the drip chamber was observed, leading to increases in the inlet pressure to more than 150 mm Hg. All three models that demonstrated obstruction have the same vent configuration, which allows cerebrospinal fluid to accumulate close to the filter when the drip chamber is held horizontally. This feature was confirmed to be the cause of the blockage. CONCLUSION: In clinical practice, special care should be taken to avoid contact of the drip chamber vents with cerebrospinal fluid, which causes obstruction and may lead to the development of gross intracranial hypertension. Specific configurations, as identified in this testing program, are safer than others in this respect. PMID- 12590688 TI - Comparison of computed tomographic angiography with digital subtraction angiography in the diagnosis of cerebral aneurysms: a meta-analysis. AB - OBJECTIVE: To compare a novel diagnostic radiological technique, computed tomographic angiography (CTA), with the standard method, namely digital subtraction angiography (DSA), in the diagnosis of cerebral aneurysms. METHODS: A comprehensive search of the world literature on CTA was performed. Articles that reported on prospective comparisons of CTA and DSA in the evaluation of patients suspected of harboring cerebral aneurysms were selected for data extraction. Suitable statistical methods were applied to the extracted data for meta analysis. RESULTS: Twenty-one references met the criteria for use in the meta analysis. Unweighted calculations based on data for 1251 patients resulted in a sensitivity of 0.933 (93.3%; range, 75.4-100%) and a specificity of 0.878 (87.8%; range, 0-100%). When the studies were weighted for the number of patients in each study, the sensitivity decreased slightly, to 0.927 (92.7%), and the specificity decreased more substantially, to 0.772 (77.2%). CONCLUSION: On the basis of this meta-analysis, DSA remains the standard method. However, many who use CTA have reported it to be as good as or better than DSA in the diagnosis and treatment of cerebral aneurysms, as well as being of less risk and discomfort to their patients and easier and less expensive to perform. PMID- 12590689 TI - Clinical prospects for neural grafting therapy for hippocampal lesions and epilepsy. AB - OBJECTIVE: Hippocampal lesions and epilepsy may be potential clinical targets for neural grafting. We hypothesized that neural grafting could be a restorative therapy either acutely, adding unformed neural elements, or chronically, treating postlesioning epilepsy. The goal of this review was to assess the clinical reality of this hypothesis of neural grafting and to determine the problems that remain to be resolved before grafting can be applied clinically. METHODS: We quantitatively defined graft integration within the host, on a cellular basis, by directly assessing survival of the transplanted neurons, graft cell dispersion and migration, neuronal differentiation and development, and establishment of appropriate local and long-distance synaptic connectivity. RESULTS: Embryonic hippocampal suspension grafts demonstrate excellent survival rates (20-80%). Embryonic axons exhibit extensive, appropriate, local and long-distance connectivity, can facilitate reconstruction of excitatory and inhibitory cortical circuitry, and can prevent the formation of aberrant circuitry. Immature neural stem cells demonstrate lesser degrees of integration, likely because of a paucity of positional cues in the lesioned brain for the differentiation of stem cells into region-specific neuronal phenotypes. Labeled grafted cells may be selectively and noninvasively removed from the host with triggerable stealth toxins, for the late treatment of unanticipated graft problems. CONCLUSION: Neural grafting with appropriate embryonic neurons may provide significant clinical benefits. However, embryonic cell availability is severely limited, and alternative sources of cells, such as stem cells, require significant additional research into the induction and maintenance of neuronal commitment and the ability of the cells to form functional synaptic connections in vivo. PMID- 12590690 TI - Duplication of the abducens nerve at the petroclival region: an anatomic study. AB - OBJECTIVE: During its course between the brainstem and the lateral rectus muscle, the abducens nerve usually travels forward as a single trunk, but it is not uncommon for the nerve to split into two branches. The objective of this study was to establish the incidence and the clinical importance of the duplication of the nerve. METHODS: The study was performed on 100 sides of 50 autopsy materials. In 10 of 11 cases of duplicated abducens nerve, colored latex was injected into the common carotid arteries and the internal jugular veins. The remaining case was used for histological examination. RESULTS: Four of 50 cases had duplicated abducens nerve bilaterally. In seven cases, the duplicated abducens nerve was unilateral. In 9 of these 15 specimens, the abducens nerve emerged from the brainstem as a single trunk, entered the subarachnoid space, split into two branches, merged again in the cavernous sinus, and innervated the lateral rectus muscle as a single trunk. In six specimens, conversely, the abducens nerve exited the pontomedullary sulcus as two separate radices but joined in the cavernous sinus to innervate the lateral rectus muscle. In 13 specimens, both branches of the nerve passed beneath the petrosphenoidal ligament. In two specimens, one of the branches passed under the ligament and the other passed over it. In one of these last two specimens, one branch passed over the petrosphenoidal ligament and the other through a bony canal formed by the petrous apex and the superolateral border of the clivus. In all of the specimens, both branches were wrapped by two layers: an inner layer made up of the arachnoid membrane and an outer layer composed of the dura during its course between their dural openings and the lateral wall of the cavernous segment of the internal carotid artery. This finding was also confirmed by histological examination in one specimen. CONCLUSION: Double abducens nerve is not a rare variation. Keeping such variations in mind could spare us from injuring the VIth cranial nerve during cranial base operations and transvenous endovascular interventions. PMID- 12590691 TI - Chronic endothelin antagonism restores cerebrovascular function in diabetes. AB - OBJECTIVE: Diabetes profoundly alters vascular function and is a risk factor for cerebrovascular disease. Diabetes increases myogenic tone and decreases responsiveness to adenosine triphosphatase (ATP)-sensitive K(+) (K(ATP)) channel openers and endothelium-dependent vasodilators. The mechanism(s) by which diabetes impairs cerebrovascular function remain obscure. In the present study, the effects of the potent vasoactive peptide endothelin-1 on myogenic tone and endothelium-dependent and potassium channel-mediated vasodilation in middle cerebral arteries from diabetic and nondiabetic rats were investigated. METHODS: Twenty-eight Wistar rats were divided into four experimental groups (n = 7 per group): control (C), control treated with bosentan (an endothelin A/B receptor antagonist) (CB), diabetic (D), and diabetic bosentan-treated (DB). Diabetes was induced with streptozotocin (D and DB groups), after which chronic bosentan treatment was initiated (CB and DB groups). Middle cerebral arteries were mounted in a pressure myograph, and myogenic responses were recorded. In addition, endothelium-dependent and -independent responses and the effects of the K(ATP) channel opener pinacidil were examined. RESULTS: Cerebral arteries from the diabetic and nondiabetic rats constricted in response to graded pressure increases. Maximum myogenic responses (percent constriction at 60 mm Hg) were significantly greater in the D group (38 +/- 3% versus 25 +/- 3% in C; P < 0.02). The enhanced myogenic tone in the D group was completely prevented by bosentan treatment (DB, 23 +/- 5% versus D; P < 0.003) without an effect on the CB group. In addition, bosentan treatment improved endothelium-dependent vasomotion and improved K(ATP)-mediated vasodilation in the DB group (P < 0.001). CONCLUSION: These data describe, for the first time, the interaction between endothelin-1, myogenic tone, and endothelial function in diabetes. Chronic endothelin antagonism restores cerebrovascular function in this model of diabetes and has global implications for the management of cerebrovascular disease in diabetes. PMID- 12590692 TI - Effects of a nitric oxide donor on and correlation of changes in cyclic nucleotide levels with experimental vasospasm. AB - OBJECTIVE: Vasospasm after subarachnoid hemorrhage (SAH) may result from hemoglobin-mediated removal of nitric oxide (NO) from the arterial wall. We tested the ability of the long-acting, water-soluble, NO donor (Z)-1-[N-(2 aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-1,2-diolate (DETA/NO), delivered via continuous intracisternal infusion, to prevent vasospasm in a nonhuman primate model of SAH. METHODS: First, vasorelaxation in response to DETA/NO was characterized in vitro by using monkey basilar artery rings under isometric tension. Next, monkeys were randomized to undergo angiography, unilateral SAH, and no treatment (SAH only, n = 4) or treatment with DETA/NO (1 mmol/L, 12 ml/d, n = 4) or decomposed DETA/NO (at the same dose, n = 4). Vasospasm was assessed by angiography, which was performed on Day 0 and Day 7. Levels of cyclic adenosine monophosphate and cyclic guanosine monophosphate (cGMP) were measured in cerebral arteries on Day 7. RESULTS: DETA/NO produced significant relaxation of monkey arteries in vitro, which reached a maximum at concentrations of 10(-5) mol/L. In monkeys, angiography demonstrated significant vasospasm of the right intradural cerebral arteries in all three groups, with no significant difference in vasospasm among the groups (P > 0.05, analysis of variance). The ratios of cGMP or cyclic adenosine monophosphate levels in the right and left middle cerebral arteries were not different among the groups (P > 0.05, analysis of variance). There was no significant correlation between arterial cGMP contents and the severity of vasospasm. CONCLUSION: DETA/NO did not prevent vasospasm. There was no correlation between the severity of vasospasm and cyclic adenosine monophosphate and cGMP levels in the cerebral arteries. These results suggest that events downstream of cyclic nucleotides may be abnormal during vasospasm. PMID- 12590693 TI - From microscopic to astronomic, the legacy of Carl Zeiss. AB - IN 1846, CARL ZEISS established a workshop to make lenses for microscopes, cameras, binoculars, and astronomical observatories. He was a master craftsman and was intolerant of any flaw, destroying microscopes with only minor inaccuracies. His relentless pursuit of perfection brought him into contact with a brilliant physicist, Ernst Abbe. Zeiss combined Abbe's new optical laws with his own technical skills; together, they created a colossus. Their company came to be known not only for exquisite technical standards but also for labor relations that were and remain progressive. The development of microneurosurgery was aided by the active participation of Carl Zeiss, Inc. The history of this company provides a lesson in the power of entrepreneurship and the benefits to humanity that can accrue from a fruitful partnership between medical science and industry. PMID- 12590694 TI - Five early accounts of phantom limb in context: Pare, Descartes, Lemos, Bell, and Mitchell. AB - PHANTOM LIMB WAS described long before American physician and surgeon Silas Weir Mitchell coined the term and drew attention to the disorder in the 1860s. The early descriptions of Ambroise Pare, Rene Descartes, Aaron Lemos, Charles Bell, and then Mitchell of this strange consequence of amputation are presented in historical and cultural context. These five men described phantom limbs for various reasons. They also differed when it came to explaining and dealing with these illusory sensations. The rich history of phantom limbs can begin to be appreciated by viewing the contributions of these individuals in perspective and by realizing that their writings represent only a fraction of what was published about phantom limbs more than 130 years ago. PMID- 12590695 TI - Fetal radiation doses for model C gamma knife radiosurgery. AB - OBJECTIVE: To assess fetal radiation doses in a pregnant patient undergoing gamma knife stereotactic radiosurgery (GKRS) and to quantify the components of extracranial radiation doses by means of phantom measurements. METHODS: A patient in her 25th week of pregnancy received GKRS with the Model C gamma knife for a solitary metastatic melanoma of the brain. A single isocenter with the 14-mm collimator and trunnions was used to deliver 20 Gy to the lesion defined to the 80% isodose line. Fetal radiation doses were assessed with phantom measurements before GKRS and then measured during GKRS. In addition, doses to the lateral canthi, thyroid, sternum, and pelvis were monitored with thermoluminescent dosimeters in 20 consecutive patients during their GKRS. Phantom measurements were also performed to identify extracranial radiation doses to these structures. RESULTS: Fetal radiation doses were 0.31, 0.20, and 0.15 cGy to the top, middle, and bottom of the uterus, respectively. These approximately corresponded to 0.01% of the maximum tumor dose of 25 Gy. The mean extracranial doses in 20 patients were 36.9, 5.8, 3.3, and 0.6 cGy to the lateral canthi, thyroid, sternum, and pelvis, respectively. Phantom measurements supported the results obtained from the patient measurements and further refined estimates of doses to extracranial sites. These measurements were also compared with those of a previously reported study with the use of the Model U. CONCLUSION: Measured radiation doses in a pregnant patient to extracranial sites, including those to the fetus, were very low. We think that GKRS is a safe treatment, particularly with the Model C, and could be recommended to carefully selected patients with brain metastases who are in the second and third trimester of pregnancy. PMID- 12590696 TI - Development of a pontine cyst after endovascular coil occlusion of a basilar artery trunk aneurysm: case report. AB - OBJECTIVE AND IMPORTANCE: Cyst formation within the brain parenchyma after endovascular coil occlusion of an intracranial aneurysm is a previously undescribed occurrence. We describe a 70-year-old woman who presented with a symptomatic pontine cyst 1 year after uncomplicated stenting and Guglielmi detachable coil occlusion of an unruptured basilar artery trunk aneurysm. CLINICAL PRESENTATION: A 70-year-old woman presented with an episode of transient dysequilibrium and gait difficulty. Magnetic resonance imaging and cerebral angiography demonstrated a 15-mm distal basilar artery trunk aneurysm. Endovascular stenting and coil occlusion of the aneurysm were performed without technical complications. One year after the initial treatment, the patient developed progressive dysarthria, dysphagia, diplopia, and left hemiparesis. A large pontomesencephalic cyst adjacent to the coiled basilar aneurysm was identified on magnetic resonance imaging scans. INTERVENTION: A subtemporal craniotomy and decompression of the pontomesencephalic cyst were performed. The patient's symptoms of brainstem dysfunction improved temporarily but recurred within 2 months, necessitating reoperation for cyst drainage and placement of a cyst-peritoneal shunt. CONCLUSION: Intra-axial cyst formation after stenting and endovascular occlusion of an intracranial aneurysm is an unusual occurrence and should be considered in the differential diagnosis of new neurological deficits after endovascular treatment. The pathophysiological mechanism of cyst formation in this case is not known. PMID- 12590697 TI - Isolated trigeminal nerve sarcoid granuloma mimicking trigeminal schwannoma: case report. AB - OBJECTIVE AND IMPORTANCE: Sarcoidosis most commonly presents as a systemic disorder. Infrequently, sarcoidosis can manifest itself in the central nervous system, with granulomas involving the leptomeninges and presenting with facial nerve weakness. Sarcoid of the trigeminal nerve is exceedingly rare and can mimic trigeminal schwannoma. We review the literature on sarcoid granulomas of the trigeminal nerve and compare their radiological features with the more common schwannoma. CLINICAL PRESENTATION: A 33-year-old woman presented with a history of left-sided facial pain and numbness for 11 months, which was presumed to be trigeminal neuralgia. A trial of carbamazepine had been unsuccessful in relieving the facial pain. Her neurological examination revealed decreased facial sensation in the V(1)-V(2) distribution. Preoperative magnetic resonance imaging demonstrated a contrast-enhancing mass centered in the left cavernous sinus with extension along the cisternal portion of the left trigeminal nerve. INTERVENTION: The patient underwent a left frontotemporal orbitozygomatic craniotomy with intraoperative neurophysiological monitoring of Cranial Nerves III, V, and VI and image guidance for subtotal microsurgical resection of what appeared, grossly and on frozen section, to be a neurofibroma. The final pathology report, however, revealed a sarcoid granuloma of the trigeminal nerve. CONCLUSION: The differential diagnosis of contrast-enhancing lesions in the lateral wall of the cavernous sinus should include inflammatory conditions such as sarcoidosis. We recommend that surgery for biopsy or decompression be used only for those patients in whom a diagnosis cannot be confirmed with noninvasive testing. If surgery is performed, intraoperative frozen pathology is very useful in guiding the extent of resection. PMID- 12590698 TI - Delayed ischemic deficit after resection of a large intracranial dermoid: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: A unique case of delayed ischemic deficit after resection of a large intracranial dermoid is presented. CLINICAL PRESENTATION: A 23-year-old woman, 36 hours after the uneventful gross total resection of a large intracranial dermoid cyst, slowly developed a progressive mixed aphasia and right hemiparesis. Magnetic resonance imaging and magnetic resonance angiography revealed small infarcts of the left putamen and temporal-occipital junction and a vasospastic tapering of the left M1 segment. INTERVENTION: Angiography confirmed severe vasospastic tapering of the left M1 and M2 segments. Endovascular treatment successfully restored flow in the left superior division. However, the initial attempt at low-pressure dilation of the inferior division led to vessel rupture. Seven months after reoperation for emergent trapping of the M1 segment, the patient made an excellent recovery, with only mild right-hand incoordination. CONCLUSION: Ruptured dermoid cysts are a risk for early and delayed cerebral ischemia, and endovascular treatment of dermoid-encased vessels may carry a higher risk for rupture. PMID- 12590699 TI - Cobb syndrome in an infant: treatment with endovascular embolization and corticosteroid therapy: case report. AB - OBJECTIVE AND IMPORTANCE: Cobb syndrome is a rare clinical entity characterized by the combination of a vascular skin nevus and an angioma in the spinal canal at the same metamere. We present a case report of an infant with Cobb syndrome. CLINICAL PRESENTATION: A 5-month-old girl presented with cutaneous hemangioma of the thoracolumbar region (T5-T12) and paraparesis. The infant was examined by magnetic resonance imaging and aortography and was diagnosed with Cobb syndrome. INTERVENTION: The patient received orally administered prednisolone therapy and underwent endovascular embolization of paravertebral and spinal angiomas with the use of n-butyl-2-cyanoacrylate. Her symptoms improved by combined treatment with liquid embolization and corticosteroid therapy. CONCLUSION: Although Cobb syndrome has been reported in older children, it is extremely rare in infants. To our knowledge, this is the first report of an infant with Cobb syndrome treated with endovascular embolization and corticosteroid therapy. The combined treatment with corticosteroid therapy and endovascular embolization of cutaneomeningospinal angioma in Cobb syndrome seems effective in alleviating symptoms and minimizing morbidity. PMID- 12590700 TI - Mini-transplants turning micro: how low can we go? PMID- 12590701 TI - Angiogenesis and plasticity: role of erythropoietin in vascular systems. AB - One of the principal functions of erythropoietin (EPO) is to stimulate the maturation of erythroid precursors. Yet EPO has recently been shown to modulate a host of cellular signal transduction pathways in pluripotent stem cells to perform multiple functions other than erythropoiesis. The production of EPO is tightly modulated by the loss of oxygen and the hypoxia-inducible factor 1. Once generated, EPO becomes a robust stimulus which regulates endothelial cell proliferation and migration as well as erythropoiesis and vascular resistance. Further downstream in the signal transduction cascade, EPO engages diverse cellular pathways--such as those involving Janus kinase 2, signal transducers and activators of transcription (STATs), mitogen-activated protein kinases (MAPKs), Bcl-x(L), protein kinase B, protein kinase C, and cysteine proteases--to provide "plasticity" to vascular systems through highly conserved mechanisms. EPO also has recently been demonstrated to inhibit the induction of apoptosis through two distinct components that involve the maintenance of the integrity of genomic DNA and the preservation of cellular membrane asymmetry. Recognition of the multipotential attributes of EPO for vascular systems may further the progress of the development of therapeutic strategies to delay the onset of degenerative diseases. PMID- 12590702 TI - Identification of tumor-associated HLA-ligands in the post-genomic era. AB - Over 10 years ago, the identification of the first tumor-specific T cell epitope shed light on the molecular principles underlying the phenomenon of tumor eradication by the immune system. Since then, a considerable number of different approaches for this task have been introduced and employed successfully, reflecting the growing knowledge about the cellular processes preceding antigen presentation as well as significant technical developments. This review tries to give an overview over available conventional strategies as well as current developments that utilize the potent large-scale screening tools of the post genomic era. PMID- 12590703 TI - Human colon adenocarcinoma in the SCID/CB6 radiation chimera is susceptible to adoptive transfer of allogeneic human peripheral blood mononuclear cells. AB - The aim of this study was to develop a murine model of human colon carcinoma (hCC) and to ascertain the potential of cellular immunotherapy in this model. Fragments of hCC obtained at surgery from 6 patients were transplanted under the kidney capsule of lethally irradiated CB6 mice radioprotected with severe combined immunodeficient (SCID) mice bone marrow. Tumor xenografts conserved their malignant behavior in the new environment, invading the mouse kidney parenchyma and expanding into the peritoneal cavity and adjacent tissues. Their growth was typically exponential, and they expanded to dimensions that allowed their subsequent fragmentation and passage to further preconditioned mice. Human carcinoembryonic antigen (hCEA) was detected on the implanted tumor and at occasionally spontaneous lung metastases. Most significantly, high levels of this tumor marker were detected in the sera of tumor-bearing mice, providing a useful tool, which allowed long-term experiments, monitoring of tumor progression, and its response to some treatment modalities. For instance, complete resection of the transplanted tumors, by means of nephrectomy, resulted in the disappearance of hCEA from mice sera within 2 weeks. Similarly, adoptive transfer of allogeneic human peripheral blood mononuclear cells (PBMC) into the peritoneum of tumor bearing mice, resulted in their rapid engraftment, infiltration of tumor mass, and a significant drop of hCEA levels in mice serum, accounting for inhibition of tumor growth. We suggest that this novel model of human colon carcinoma affords the opportunity for in vivo evaluation of different preclinical treatment modalities, particularly, those involving manipulation with immune effector cells. PMID- 12590704 TI - 4-1BB ligand stimulation enhances myeloid dendritic cell maturation from human umbilical cord blood CD34+ progenitor cells. AB - In humans, at least two subsets of dendritic cells (DCs) are identified on the basis of differential surface expression of CD11c antigens. CD11c(+) and CD11c(-) cells are respectively of myeloid and lympholoid origin and functionally distinct, eliciting inflammatory and tolerant T cell responses. We investigated whether 4-1BB ligand (4-1BBL), a member of the tumor necrosis factor (TNF) family, is involved in the maturation process to mature myeloid DCs during in vitro DC differentiation from immature DCs derived from human umbilical cord blood (CB) CD34(+) progenitor cells. Enhanced levels of CD11c as well as immunostimulatory molecules such as CD86, MHC class II, and 4-1BBL were induced in response to 4-1BBL stimulation. These changes were accompanied by noticeable morphological transition from nonadherent to adherent myeloid-like DCs. Stimulation of 4-1BBL on DCs with 4-1BB-Fc or with 4-1BB-transfected Jurkat cells resulted in acquisition of capacity for the immature DCs to produce interleukin 12 (IL-12). This suggests that 4-1BBL may be an important mediator for maturation of CD11c(+) myeloid DCs, information of possible relevance for the design of DC based vaccines with enhanced activity. PMID- 12590705 TI - Hematopoietic stem cell functional failure in interleukin-2-deficient mice. AB - The effects of interleukin-2 (IL-2) deficiency on hematopoiesis were tested by measuring cellular compositions in peripheral blood, spleen, thymus, and bone marrow of 3- to 5-month-old gene-targeted Il2 null (Il2(-/-)) mice using the Advia 120 Hematology system and fluorescence-activated cell staining (FACS). Il2( /-) mice developed hematological failure and autoimmune responses, showing variable but significant degrees of anemia, lymphocytopenia, thrombocytopenia, splenomegaly, thymus involution, and weight loss. Surprisingly, Il2(-/-) mice had normal numbers of bone marrow cells (BMCs) with increased numbers of Lin( )Kit(+)Sca1(+)CD34(-) and Lin(-)Kit(+)Sca1(+)CD34(+) cells that are normally associated with hematopoietic stem cells (HSCs) and progenitor cells. Day-12 colony-forming units-spleen cells were slightly reduced in Il2(-/-) mice. When Il2(-/-) and Il2(+/+) mice were compared for long-term HSC function in vivo in the competitive repopulation assay, BMCs from Il2(-/-) donors had 10- to 20-fold less HSC repopulating ability, which affected both myeloid and lymphoid cell lineages. Thus, HSCs from Il2(-/-) mice can proliferate normally but are functionally defective for reconstituting lethally irradiated recipients. PMID- 12590706 TI - Homing of purified murine lymphohematopoietic stem cells: a cytokine-induced defect. AB - This study was designed to establish a direct homing assay using purified lineage negative Sca-1-positive (Lin(-) Sca(+)) murine bone marrow cells and to evaluate the effects of cytokines on homing. C57BL/6 Lin(-) Sca(+) marrow stem cells were labeled with 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and then injected by tail vein into untreated C57BL/6 mice. Marrow was harvested at various times after cell infusion and analyzed on a high-speed MoFlo cell sorter for fluorescent positive events, using a large event analysis, with at least 16 million total events analyzed. We have shown that homing of Lin(-) Sca(+) cells plateaus by 1 h, and at 3 h post-infusion is linear between 50,000 and 1,000,000 infused cells. This forms a base for a homing assay in which 250,000 CFDA-SE labeled Lin(-) Sca(+) marrow cells are infused and then recovered from marrow 3 h later, followed by a large-event fluorescence-activated cell sorting (FACS) analysis. We found that 7.45-9.32% of infused cells homed and that homing of stem cells cultured for 48 h in interleukin-3 (IL-3), IL-6, IL-11, and steel factor cultured cells was defective when compared to noncultured cells. Exposure of marrow stem cells to IL-3, IL-6, IL-11, and steel factor induces a stem cell homing defect, which probably underlies the engraftment defect previously characterized under these conditions. PMID- 12590708 TI - Influence of ex vivo expansion and retrovirus-mediated gene transfer on primary T lymphocyte phenotype and functions. AB - To modulate alloreactivity after hematopoietic stem cell (HSC) transplantation, suicide gene-expressing donor T cells can be administered with an allogeneic T cell-depleted HSC graft. Immune competence of such cells is a critical issue. We have examined the impact of our ex vivo gene transfer protocol (12-day culture period including CD3/IL-2 activation, retrovirus-mediated gene transfer, and G418 based selection) on the phenotype and functional properties of gene-modified cells (GMC). GMC were compared with control cells that had been cultured in parallel with GMC, but nontransduced and nonselected, as well as with peripheral blood mononuclear cells (PBMC). Our data show that phenotypical modifications are similar in control cells and GMC, demonstrating that alterations result from the 12-day culture rather than from the transduction and/or selection process itself. Such modifications include a reversal of CD4/CD8 ratio, activated phenotype (increased expression of CD45RO, CD95, and HLA-DR), and acquisition or increased expression of co-stimulatory molecules (CD80, CD86, and CD40). This led to an enhanced allostimulating potential of GMC, as compared with resting T cells, when used as stimulating cells in mixed lymphocyte reactions. Conversely, when using them as responder cells in mixed lymphocyte reactions, GMC exhibited a rapid loss of alloreactivity that resulted both from culture-dependent and from transduction and/or selection-dependent events. In conclusion, the retrovirus-mediated gene transfer can be associated with major phenotypical and functional alterations that could have strong clinical implications (increased immunogenicity, reduced anti-leukemic effect). Thus, future T cell expansion protocols should try to improve not only cell expansion or gene transfer efficiency, but also T cell functions. PMID- 12590707 TI - Src kinase, but not the src kinase family member p56lck, mediates stromal cell derived factor 1alpha/CXCL12-induced chemotaxis of a T cell line. AB - Stromal cell-derived factor 1 (SDF-1/CXCL12) is believed to mediate migration of leukocytes. To explore potential mechanisms, we evaluated the signal transduction pathways activated by SDF-1 in the Jurkat T cell line. Src kinase was phosphorylated and mitogen-activated protein kinase/extracellular signal regulated kinase (MAPK/ERK) activated in a time-related fashion after SDF-1 stimulation. Chemotaxis of Jurkat cells was partially blocked by pretreatment with the src kinase inhibitor PP2 in a dose-dependent manner. Wiskott-Aldrich Syndrome Protein (WASP) regulates actin polymerization and cytoskeletal organization in T cells. We found WASP complexed to activated src after SDF-1 stimulation, suggesting a possible interacting role for src kinase and WASP in mediating SDF-1 action. J.CaM1.6 cells, which have lost expression of the src kinase p56(lck) (lck), responded to chemotaxis induced by SDF-1 as well as the parental Jurkat cells. Because J.CaM1.6 cells respond as well as the parental cells to SDF-1 in terms of ERK activation and tyrosine phosphorylation of WASP after SDF-1 stimulation, it appears that src kinase, but not the src kinase family member lck, mediates chemotaxis of Jurkat cells in response to SDF-1 induction and that src kinase may link with WASP in this effect. PMID- 12590710 TI - Early apoptosis largely accounts for functional impairment of CD34+ cells in frozen-thawed stem cell grafts. AB - Quality assessment of stem cell grafts is usually performed by flow cytometric CD34(+) enumeration or assessment of clonogenic output of fresh material. Previously, we identified the occurrence of early apoptosis, not detectable with the permeability marker 7-amino actinomycin D (7-AAD), in purified frozen-thawed CD34(+) cells, using the vital stain Syto16. Syto(high)/7-AAD(-) cells were defined as viable, Syto16(low)/7-AAD(-) cells as early apoptotic and Syto16(low)/7-AAD(+) as dead. This was confirmed in a subsequent study using frozen-thawed transplants of lymphoma patients. In the present study on grafts from multiple myeloma and lymphoma patients, we investigated the functional consequences of the early apoptotic process. The mean Syto16-defined viability was 41 and 42%, respectively, for both graft groups, compared to 78% and 72%, respectively, using 7-AAD only. The established early apoptosis marker annexin V missed roughly 50% of the early apoptosis detected with Syto16. In contrast, viability of CD34(+) cells in nonmanipulated whole blood transplants from a matched group of lymphoma patients, after 72 h of storage at 4 degrees C, was more than 90%, even with the Syto16 assay. CFU recovery (median 26-33%) after cryopreservation matched CD34(+) recovery after Syto16, but not 7-AAD correction. In contrast, colony-forming unit (CFU) recovery in the whole blood transplant was close to 100%. Furthermore, early apoptotic CD34(+) cells had lost migratory ability toward stromal cell derived factor-1alpha (SDF-1alpha). The establishment of a Syto16(high)/7-AAD(-) proportion of CD34(+) cells offers a new approach for a more correct determination of the number of viable nonapoptotic CD34(+) cells in stem cell grafts. Further development of this assay should allow its incorporation into the routine CD34(+) assessment of post-thawed samples in clinical flow cytometry laboratories. PMID- 12590709 TI - Acute promyelocytic leukemia M3: cytomorphologic, immunophenotypic, cytogenetic, and molecular variants. AB - Acute promyelocytic leukemia (APL) M3 is an acute myeloid leukemia (AML) subtype characterized by proliferation of malignant promyelocytes with mature myeloid immunophenotype and the translocation t(15;17)(q22;q11), which results in the fusion of retinoic acid receptor-alpha (RARalpha) gene on chromosome 17 and the gene PML on chromosome 15. There are three M3 morphologic variants: the typical hypergranular form and the microgranular and basophilic variants. Although most leukemic cells in M3 patients express t(15;17), other cytogenetic abnormalities have also been reported. Also, there are three molecular variants of the PML/RARalpha transcript (bcr1, bcr2, bcr3). Blasts had typical hypergranular appearance (13 patients) with a mature myeloid immunophenotype (HLA-DR( ),CD13(+), and/or CD33(+)) (10 patients) in the majority of patients with M3 followed in this study. The typical translocation [t(15;17)(q22;q11)] was detected by cytogenetic analysis in 5 M3 patients, but PML/RARalpha was positive in 13 out of 15 patients, as assessed by RT-PCR (8 patients with bcr1 and 5 with bcr3 subtype). Cytogenetic diversity was found in three patients (1 with t(17;17), 1 with +8, and 1 with add (7)(q22); -7; +8). According to many studies, leukemic cell heterogeneity in APL influences the clinical outcome of disease. The analysis of certain leukemic cell characteristics on the clinical outcome in our study revealed that patients with bcr3 had shorter medians of first remission and survival in comparison to patients with the bcr1 isoform of PML/RARalpha. Also, the clinical relapse of disease in 4 APL patients with reverted PML/RAR alpha positivity is consistent with the view that detection of PML/RARalpha by RT RCR in patients in remission implies a poor prognosis. On the contrary, lack of detection of PML/RARalpha by RT-PCR at least three times is a sign of long remission and survival. PMID- 12590711 TI - Hematopoietic reconstitution of irradiated, stem cell-injected mice: early dynamics of restoration of the cell lineages of the spleen and bone marrow. AB - Hematopoietic stem cells, numbering approximately 1/100,000 cells in mammalian bone marrow, are capable of complete hematopoietic and immune reconstitution upon injection into a myeloablated host. The present study aimed to analyze the earliest events in reconstitution of lethally irradiated, host murine bone marrow and spleen, after injecting purified Thy 1(lo)Lin(-)Sca-1(+) stem cells. Thy 1(lo)Lin(-)Sca-1(+) cells were isolated by fluorescence-activated cell sorting (FACS) from the bone marrow of 4-week-old C57BL(Thy1.1, Ly5.1) mice and injected into preirradiated, syngeneic hosts. These stem cells were also injected into congenic hosts, i.e., C57BL(Thy1.2, Ly5.2), and confirmed the donor origin of hematopoietic cells in the reconstituted host mice. Hematologically stained smears of the spleen and bone marrow of stem cell-injected recipients were prepared at 11, 14, 17, 21, 24, and 28 days after stem cell injection, and nucleated erythroid cells, mature granulocytes, and their myeloid precursors, monocytes, and large and small lymphocytes were recorded as a proportion of all nucleated cells in each organ at each time interval. The results indicated that in the earliest post stem cell injection intervals, both organs were predominantly erythroid and myeloid. Only at the later intervals did both organs show high proportions of large lymphoid cells and their progeny, small lymphocytes. Thus, early (<1 month) dynamics of hematopoietic reconstitution after transplantation of purified hematopoietic stem cells, is cell lineage specific. PMID- 12590713 TI - Human mesenchymal stem cells are not of donor origin in patients with severe aplastic anemia who underwent sex-mismatched allogeneic bone marrow transplant. AB - Stromal defects are part of the etiology of severe aplastic anemia (SAA), and hematopoietic engraftment is poor in unrelated and mismatched transplant. Therefore, we wanted to find out whether human mesenchymal stem cells (MSC) are partly of donor origin in patients with SAA years after successful bone marrow transplant (BMT). Three SAA patients 3, 5, and 8 years after BMT (cyclophosphamide, ATG) with bone marrow from an HLA-identical sibling donor of the opposite sex were investigated. MSC were grown from patients' bone marrow aspirates according to Caplan et al. The number of MSC that were isolated from SAA bone marrow post transplant was about 10 times lower than in normal controls. Primary cultures of adherent MSC and passage-one cells were analyzed by dual color interphase fluorescence in situ hybridization (FISH) analysis using centromere-specific DNA probes for X and Y chromosome. FISH did not show any clear evidence of donor cells in the adherent MSC: In all cases, less than 0.5% of nuclei showed a donor-type signal pattern that is well within assay limits. In a female patient, the absence of male donor cells was confirmed by sensitive and quantitative, Y chromosome-specific TaqMan PCR (QYCS-PCR). In contrast, Ficoll separated hematopoietic cells from the same aspirates were greater than 90% of donor origin, as expected. In SAA, as previously found in patients with lysosomal and peroxisomal storage disease, bone marrow MSC remain host-derived despite successful hematopoietic engraftment years after allogeneic BMT. PMID- 12590712 TI - Molecular methods for detection and quantification of myeloma cells after bone marrow transplantation: comparison between real-time quantitative and nested PCR. AB - Multiple myeloma is characterized by malignant plasma cell-infiltration of bone marrow. Treatment with high-dose therapy results in a high rate of clinical remissions, but almost all patients ultimately relapse. Clinical staging and detection of relapse are limited in sensitivity. Therefore, we established molecular methods based on the highly clone-specific CDR regions of the immunoglobulin VH locus for sensitive and specific detection of residual myeloma cells after bone marrow transplantation. VDJ rearrangements were identified using a set of VH primers and a JH primer. Clone-specific rearrangements were detected by comparison with germ-line sequences. With the nested PCR approach, first-round amplification with the consensus primers was done followed by second amplification with myeloma-specific primers. The real-time quantitative PCR was performed using a myeloma-specific forward primer in combination with a JH consensus TaqMan probe and reverse primer. Sensitivity was tested using dilutions of myeloma cell lines into mononuclear cells. Nested PCR had a sensitivity of 10( 6) and TaqMan PCR of 10(-4) to 10(-5). Specificity was determined by testing different cell lines and patients' probes. These results were confirmed by follow up of 2 patients after allogeneic transplantation with dose-reduced conditioning. Molecular methods are very sensitive and specific tools for follow up of myeloma patients after allogeneic transplantation. By using the quantitative approach, it is possible to see kinetics of bone marrow tumor load, which can be used to guide therapeutic decisions like donor leukocyte infusions (DLI). PMID- 12590714 TI - The "cloning effect": insufficient evidence for pluripotency. PMID- 12590715 TI - The role of high fluorescent reticulocytes in monitoring the aplasia outcome and optimizing the timing of peripheral blood stem cell harvesting. PMID- 12590716 TI - Immune recovery in breast cancer patients after tandem high-dose chemotherapy rescued by selected CD34+ cells. PMID- 12590718 TI - The Stretta procedure for the treatment of GERD: a registry of 558 patients. AB - PURPOSE: To evaluate gastroesophageal reflux disease (GERD) symptoms, patient satisfaction, and antisecretory drug use in a large group of GERD patients treated with the Stretta procedure (endoluminal temperature-controlled radiofrequency energy for the treatment of GERD) at multiple centers since February 1999. METHODS: All subjects provided informed consent. A health care provider from each institution administered a standardized GERD survey to patients who had undergone Stretta. Subjects provided (at baseline and follow-up) (1) GERD severity (none, mild, moderate, severe), (2) percentage of GERD symptom control, (3) satisfaction, and (4) antisecretory medication use. Outcomes were compared with the McNemar test, paired t test, and Wilcoxon signed rank test. RESULTS: Surveys of 558 patients were evaluated (33 institutions, mean follow-up of 8 months). Most patients (76%) were dissatisfied with baseline antisecretory therapy for GERD. After treatment, onset of GERD relief was less than 2 months (68.7%) or 2 to 6 months (14.6%). The median drug requirement improved from proton pump inhibitors twice daily to antacids as needed (P < .0001). The percentage of patients with satisfactory GERD control (absent or mild) improved from 26.3% at baseline (on drugs) to 77.0% after Stretta (P < .0001). Median baseline symptom control on drugs was 50%, compared with 90% at follow-up (P < .0001). Baseline patient satisfaction on drugs was 23.2%, compared with 86.5% at follow-up (P < .0001). Subgroup analysis (<1 year vs. >1 year of follow-up) showed a superior effect on symptom control and drug use in those patients beyond 1 year of follow-up, supporting procedure durability. CONCLUSIONS: The Stretta procedure results in significant GERD symptom control and patient satisfaction, superior to that derived from drug therapy in this study group. The treatment effect is durable beyond 1 year, and most patients were off all antisecretory drugs at follow-up. These results support the use of the Stretta procedure for patients with GERD, particularly those with inadequate control of symptoms on medical therapy. PMID- 12590719 TI - Laparoscopic restorative proctocolectomy for patients with ulcerative colitis. AB - BACKGROUND: Significant concern continues about the feasibility of laparoscopic restorative proctocolectomy (RP) with an ileal J pouch anal anastomosis in the surgical treatment of patients with ulcerative colitis (UC). The aim of this study was to clarify the feasibility of laparoscopic RP at a single institution where the surgical routine of laparoscopic colorectal surgery has already been established. PATIENTS AND METHODS: Between July 1994 and December 2001, 18 patients with UC underwent laparoscopic RP. The median age was 30 (range, 18-51) years, and the median follow-up was 20 (range, 5-89) months. Five trocars were placed. After the entire colon and rectum were mobilized and the vessels were divided intracorporeally, the rectum was divided with use of a laparoscopic linear stapler. A pouch anal anastomosis was fashioned with use of a double stapling technique. A diverting loop ileostomy was fashioned. RESULTS: There were no conversions to the open procedure. The median operative time and median blood loss were 360 (range, 290-500) minutes and 105 (range, 10-586) mL, respectively. Six postoperative complications occurred (wound sepsis, 2; bowel obstruction, 1; anastomotic stricture, 2; pouchitis, 1). In one patient, a bowel obstruction developed 3 months after the operation, which was managed conservatively. The median length of the hospital stay was 9 (range, 7-21) days. CONCLUSIONS: The laparoscopic RP is safe and feasible in selected patients with UC. New laparoscopic instrumentation, such as a linear stapler, and a more reliable laparoscopic coagulating and dividing tool should be designed, which would make it possible to perform this procedure more frequently in the surgical treatment of UC. PMID- 12590720 TI - Liver function disturbances following laparoscopic cholecystectomy: incidence and significance. AB - BACKGROUND: Unexplained disturbances of liver function parameters after laparoscopic cholecystectomy have been reported in a few previous studies. We aimed to assess the incidence of these disturbances, and their relation to age, sex, duration of surgery, and type of gallbladder inflammation, as well as their clinical significance. PATIENTS AND METHODS: All patients undergoing laparoscopic cholecystectomy at Dallah Hospital, Riyadh, Saudi Arabia, during the year 2001 were prospectively evaluated. In all the patients, six parameters of liver function (direct bilirubin, indirect bilirubin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyltransferase) were assessed before and 24 hours after surgery. Patients with suspected chronic liver disease or common bile duct pathology were excluded. Patients who showed more than a 100% increase in at least one parameter (group I) were compared with those who did not (group II) regarding age, sex, duration of surgery, and type of gallbladder inflammation. RESULTS: Of the patients, 67.1% showed more than a 100% increase in at least one parameter of liver function. No significant difference was noticed between the two groups regarding age, duration of surgery, or type of gallbladder inflammation. Female patients, however, showed a significantly higher incidence of liver function changes (P = .011). No clinical adverse reactions were reported in those who showed liver function changes. CONCLUSION: Changes in liver function parameters after laparoscopic cholecystectomy are common, carrying no clinical sequalae. They are more common in female patients and are not related to age, duration of surgery, or the acuteness of gallbladder inflammation. Assessment of direct bilirubin and alkaline phosphatase is sufficient for the routine evaluation of these patients postoperatively. PMID- 12590721 TI - Bare bones laparoscopy: a randomized prospective trial of cost savings in laparoscopic cholecystectomy. AB - OBJECTIVE: Rising costs and lowered reimbursements make value essential if laparoscopic cholecystectomy (LC) is to be offered to patients without condemning providers to financial loss. We hypothesize that our protocol increases this value. Once practiced, operative time, complications, and patient satisfaction compare with those of the typical method. METHODS: We prospectively randomized 50 consecutive patients equally to control or experimental LC according to our protocol. Equipment costs, operative time, conversions, complications, pain, and return to work were compared. The student's t test was used for comparisons. RESULTS: Mean disposable equipment costs were 173.00 dollars +/- 43.45 dollars and 434.42 dollars +/- 50.54 dollars for the study and control groups, respectively (P < .0001). Mean operative times were 67.26 +/- 15 and 70.60 +/- 19 minutes, respectively. CONCLUSIONS: The "bare bones" protocol is safe. It has a short learning curve, demonstrates a cost advantage over the common method, and requires no additional operative time. Pain, time to return to work, and satisfaction are equivalent. PMID- 12590722 TI - Lessons learned from ligation of indirect hernia sac: an alternative to reduction during endoscopic extraperitoneal inguinal hernioplasty. AB - BACKGROUND: During endoscopic totally extraperitoneal inguinal hernioplasty (TEP), complete reduction of the hernia sac is not always feasible because a sac extends into the scrotum or is adhesed. In such circumstances, ligation of the hernia sac appears to be a sound alternative to reduction. The present study was undertaken to evaluate the safety of sac ligation and the clinical results for patients who underwent ligation of an indirect hernia sac during TEP. PATIENTS AND METHODS: From September 1999 to July 2001, patients undergoing unilateral TEP for indirect inguinal hernia were recruited. Patients were divided into two groups. Group 1 (n = 65) underwent complete reduction of the hernia sac, whereas group II (n = 34) underwent ligation of the sac with a Vicryl suture followed by distal transection. Clinical parameters and outcome data were compared between the two groups of patients. RESULTS: Demographic features and hernia types between the two groups were comparable. The mean operative times of patients from groups I and II were 58 and 62 minutes, respectively (P = NS). Intraoperative complications occurred in two patients in group II, including a vas deferens transection and a gonadal vessel division. Comparison of the lengths of hospital stay, postoperative pain scores at rest and on coughing, rates of postoperative morbidity, and incidences of groin collection showed no significant differences between the two groups. CONCLUSIONS: Our findings highlight the importance of clear identification and protection of the vas deferens and gonadal vessels before transection of the ligated sac, particularly in patients with abundant preperitoneal adipose tissue. Ligation of an indirect hernia sac was associated with a higher incidence of intraoperative complications and should be performed with caution during TEP. PMID- 12590723 TI - Outcome of laparoscopic ventral hernia repair in correlation with obesity, type of hernia, and hernia size. AB - BACKGROUND: The morbidity and overall recovery time of ventral hernia repair can vary significantly depending on the hernia type or size and on other factors, such as the body mass index (BMI). The purpose of our study was to investigate the effects of type of hernia, hernia size, and BMI on the outcome of laparoscopic ventral hernia repair. METHODS: Fifty patients who underwent laparoscopic ventral hernia repair were retrospectively reviewed and stratified according to hernia type (incisional, IVH/primary, PVH), hernia size, and BMI. These subgroups were compared in regard to operative time, resumption of diet, hospital stay, pain control, and complication rate. RESULTS: Laparoscopic IVH repair was associated with a longer operative time (143 vs. 98 minutes, p = .02) and length of stay (2.2 vs. 0.6 days, p = .03) than PVH repair. The narcotic requirements were higher in the IVH group, but the difference did not reach statistical significance. Larger hernias were associated with a longer operative time (p = .04) and increased narcotic requirement (p = .03). The morbidity of the laparoscopic repair was not significantly affected by the hernia type or size. The BMI did not significantly alter any of the parameters examined. CONCLUSIONS: Laparoscopic repair of incisional and larger hernias is a technically demanding procedure that requires a longer operative time. In contrast to PVH repair, laparoscopic IVH repair usually cannot be performed on an outpatient basis. Surgeons need not be discouraged from recommending the laparoscopic approach for patients with large IVHs or with severe obesity because the morbidity remains low. PMID- 12590724 TI - Adult nonpalpable testis: is laparoscopy always required? AB - Laparoscopy is widely used in the diagnosis and treatment of nonpalpable testes. Some nonpalpable testes are vanishing testes. In such cases, unnecessary laparoscopic interventions can be avoided by a careful selection of cases. Between 1996 and 2001, laparoscopic intervention was applied to 107 patients with nonpalpable testes. Of the cases, 23 were bilateral and 84 were unilateral. Patients were between 19 and 27 years of age (average age, 23 years). Diagnostic ultrasonography was performed in 44 of the 84 patients with nonpalpable testes. Dimensions of the scrotal testis were determined by the Prader orchiometer method. The dimensions of the opposite scrotal testis (of the scrotal nubbin) and the abdominal testis were compared with the dimensions of 20 normal, healthy individuals' scrotal testis (control group). Results were evaluated by the Mann Whitney U test. During laparoscopy, 24 (28.5%) of the patients were found to have a vanishing testis. The vas deferens and the testicular blood vessels ended bluntly at the anterior edge of the interior inguinal ring in one patient, inside the inguinal canal in five patients, and in the scrotum in 18 patients. Among the 84 patients with nonpalpable testes, no testis was found in any of the 18 patients with palpable scrotal nubbins. The opposite scrotal testes were hypertrophic in 17 (70.8%) of 24 patients who had vanishing testis (P < .05), and they were hypertrophic in 22 (36%) of the 60 patients (P > .05) who had laparoscopically identified intraabdominal testes. We conclude that clinical and radiologic diagnosis is sufficient for adult patients with nonpalpable testicles and palpable scrotal nubbins and hypertrophic contralateral scrotal testes. Laparoscopic intervention should be applied to patients who do not have palpable scrotal nubbins. PMID- 12590725 TI - Development of a novel method of female sterilization: II. Retention of tubal screws in patients undergoing simultaneous laparoscopic sterilization. AB - BACKGROUND AND OBJECTIVES: This study was a long-term follow-up of patients in whom hysteroscopic tubal screws had been applied at the time of laparoscopic sterilization. METHODS: Tubal screw application was performed before laparoscopic Filshie clip application. Follow-up ultrasonography was arranged 3, 6, and 9 months postoperatively to confirm retention. The tubal screws were removed hysteroscopically between 12 and 20 months after sterilization under local or light general anesthesia. RESULTS: Thirty-five women agreed to take part in the study. For the purpose of analysis, these patients were divided into an initial group (cases 1-20, group A) and a later group (cases 21-35, group B). Twenty three patients had 41 tubal screws inserted (18 women had bilateral screw application). Twenty tubal screws were removed from 13 patients between 9 and 20 months after insertion, one screw remained in situ, and 20 screws had previously been extruded. Life table analysis plots demonstrated a marked but nonsignificant difference (P = .163) in the duration of tubal screw retention between the initial patients (group A) and the later patients (group B): 46.7% versus 76.9% at 6 months and 33% versus 61.5% at 12 months (P = .09 and P = .11, respectively). CONCLUSIONS: Our experience demonstrated improved application and retention with experience and refinement of the equipment; however, retention of the tubal screws, even in the later stages of development, was poor. A relatively noninvasive method of female sterilization remains the ideal, and further refinements are required. PMID- 12590726 TI - A cause for conversion of laparoscopic splenectomy: splenic candidiasis. AB - Laparoscopic splenectomy (LS) is now considered the procedure of choice for removal of the spleen in several hematologic and traumatic splenic conditions. Perisplenitis is still considered a relative contraindication. We report a rare case of isolated splenic candidiasis treated by laparoscopic splenectomy. The technical challenges of the laparoscopic approach in cases of perisplenitis are outlined. Because of dense adhesions to the diaphragm, the LS was converted to open splenectomy, and the procedure was completed through a small subcostal incision. This case report suggests the need for and effectiveness of splenectomy in patients whose fungal infection is suspected to be localized to the spleen. Laparoscopic removal of the spleen in these cases is very difficult to accomplish because of the risk for diaphragm injuries and the technical problems encountered. PMID- 12590727 TI - Urgent laparoscopic splenectomy in a morbidly obese pregnant woman: case report and literature review. AB - Laparoscopic splenectomy (LS) has undergone significant evolution since its introduction more than a decade ago. It clearly has been shown to be advantageous in comparison with open splenectomy and is considered by some to be the treatment of choice for routine uncomplicated elective splenectomy. When LS was first introduced, contraindications to the procedure included pregnancy, obesity, and splenomegaly. Both technologic advances and experience with the technique have enabled surgeons to perform LS for a growing number of indications with seemingly fewer contraindications. Here, we present a case of successful LS in a morbidly obese pregnant woman with splenomegaly and hemolysis secondary to hereditary spherocytosis. PMID- 12590728 TI - Laparoscopic "radical appendectomy" is an effective alternative to endoscopic removal of cecal polyps. AB - BACKGROUND: The endoscopic removal of cecal polyps can be complicated by hemorrhage, perforation, or incomplete resection. Laparoscopic radical appendectomy represents a safe alternative for the definitive resection and accurate pathologic evaluation of selected cecal polyps. METHODS: Patients with cecal cap polyps not involving the ileocecal valve were candidates for laparoscopic radical appendectomy. Intraoperative colonoscopy and resection of the appendix and cecum to the level of the ileocecal valve were accomplished via three midline ports. For each patient, histologic evaluation by frozen section ruled out malignancy and ensured complete resection. RESULTS: Five patients, four of whom had significant medical comorbidities, presented with large adenomatous polyps contained within the cecum. Each polyp was determined to be unresectable endoscopically; therefore, a laparoscopic radical appendectomy was performed. One patient with cirrhosis also underwent intraoperative liver ultrasonography and biopsies, which contributed to the longest operative time and hospital stay. The histologic diagnosis by frozen section was benign for each patient. The mean operative time was 95 minutes, and the mean length of hospital stay was 1.8 days. No postoperative complications were observed during a mean follow-up of 6 months. CONCLUSION: Laparoscopic "radical appendectomy" is an effective treatment for selected cecal adenomatous polyps. Our ability to resect the polyps completely and avoid a standard right hemicolectomy supports this approach. PMID- 12590729 TI - Rectal duplication cyst in an adult: the laparoscopic approach. AB - BACKGROUND: Rectal duplication cyst (RDC) is a rare congenital anomaly representing 1% to 8% of all intestinal duplications. The case presented here is the first report of the laparoscopic resection of an RDC. METHODS: We report the case of a 49-year-old white woman in whom a retrorectal cystic mass measuring 5 x 5.3 x 6 cm was diagnosed. The mass was completely resected by means of laparoscopic techniques. RESULTS: Pathologic findings revealed a cystic structure partially lined with squamous as well as respiratory- and gastrointestinal-type epithelium. Muscularis propria was identified in the outer portions of the wall of the specimen. No atypia or malignancy was identified. The overall findings were consistent with an RDC. CONCLUSIONS: Laparoscopic resection constitutes an excellent and patient-friendly approach to the management of large adult cystic duplication of the rectum. PMID- 12590730 TI - Laparoscopic repair of diaphragmatic Morgagni hernia. AB - A novel case of laparoscopic repair of a diaphragmatic hernia through the foramen of Morgagni in a 5-year-old boy is reported. The patient had a prompt and complete recovery, with no evidence of recurrence noted at 1 year after surgery. Laparoscopic repair is considered to be a suitable and safe procedure for the treatment of Morgagni hernia. PMID- 12590731 TI - The perspective effect of wide-angle lenses in laparoscopes. AB - PURPOSE: To evaluate the effect of perspective distortion of wide-angle lenses in laparoscopes on hand-eye coordination during endoscopic manipulation. METHODS: Sixteen subjects repeatedly performed a standardized positioning task in a pelvi trainer under two conditions. The subjects had no prior experience with endoscopic manipulation. In one condition, a wide-angle lens with considerable perspective distortion was used; in the other, a telephoto lens without perspective distortion was used. Task time and number of errors were measured. RESULTS: Task time and the number of errors did not significantly increase in the condition with a wide-angle lens. CONCLUSIONS: The perspective effect did not influence task performance in endoscopic manipulation in this experiment. Subjects indicated that they even preferred the wide-angle lens because its extreme perspective improved their perception of depth. PMID- 12590732 TI - ORP3 splice variants and their expression in human tissues and hematopoietic cells. AB - ORP3 is a member of the newly described family of oxysterol-binding protein (OSBP)-related proteins (ORPs). We previously demonstrated that this gene is highly expressed in CD34(+) hematopoietic progenitor cells, and deduced that the "full-length" ORP3 gene comprises 23 exons and encodes a predicted protein of 887 amino acids with a C-terminal OSBP domain and an N-terminal pleckstrin homology domain. To further characterize the gene, we cloned ORP3 cDNA from PCR products and identified multiple splice variants. A total of eight isoforms were demonstrated with alternative splicing of exons 9, 12, and 15. Isoforms with an extension to exon 15 truncate the OSBP domain of the predicted protein sequence. In human tissues there was specific isoform distribution, with most tissues expressing varied levels of isoforms with the complete OSBP domain; while only whole brain, kidney, spleen, thymus, and thyroid expressed high levels of the isoforms associated with the truncated OSBP domain. Interestingly, the expression in cerebellum, heart, and liver of most isoforms was negligible. These data suggest that differential mRNA splicing may have resulted in functionally distinct forms of the ORP3 gene. PMID- 12590733 TI - Processing of filamentous bacteriophage virions in antigen-presenting cells targets both HLA class I and class II peptide loading compartments. AB - Virions of filamentous bacteriophage fd are capable of displaying multiple copies of peptide epitopes and generating powerful immune responses to them. To investigate the antigen processing mechanisms in human B cell lines used as antigen presenting cells, the major coat protein (pVIII) in intact virions was fluorescently labeled, and its localization in various intracellular compartments was followed using confocal microscopy. We show that the virions were taken up and processed to yield peptides that reach both the major histocompatibility complex (MHC) class II compartment and the endoplasmic reticulum. Moreover, when exposed to bacteriophages displaying a cytotoxic T lymphocyte (CTL) epitope from the reverse transcriptase of human immunodeficiency virus type-1 (HIV-1), B cells were lysed by specific cytotoxic lymphocytes. This confirms that filamentous bacteriophage virions are capable of being taken up and processed efficiently by MHC class I and class II pathways, even in nonprofessional antigen presenting cells. These remarkable features explain, at least in part, the unexpected ability of virions displaying foreign T-cell epitopes to prime strong T-helper dependent CTL responses. These findings have important implications for the development of peptide-based vaccines, using filamentous bacteriophage virions as scaffolds. PMID- 12590734 TI - The mouse sulfate anion transporter gene Sat1 (Slc26a1): cloning, tissue distribution, gene structure, functional characterization, and transcriptional regulation thyroid hormone. AB - Sulfate (SO(4)(2-)) is required for bone/cartilage formation and cellular metabolism. sat-1 is a SO(4)(2-) anion transporter expressed on basolateral membranes of renal proximal tubules, and is suggested to play an important role in maintaining SO(4)(2-) homeostasis. As a first step towards studying its tissue specific expression, hormonal regulation, and in preparation for the generation of knockout mice, we have cloned and characterized the mouse sat-1 cDNA (msat-1), gene (sat1; Slc26a1) and promoter region. msat-1 encodes a 704 amino acid protein (75.4 kDa) with 12 putative transmembrane domains that induce SO(4)(2-) (also oxalate and chloride) transport in Xenopus oocytes. msat-1 mRNA was expressed in kidney, liver, cecum, calvaria, brain, heart, and skeletal muscle. Two distinct transcripts were expressed in kidney and liver due to alternative utilization of the first intron, corresponding to an internal portion of the 5'-untranslated region. The Sat1 gene (~6 kb) consists of 4 exons. Its promoter is ~52% G + C rich and contains a number of well-characterized cis-acting elements, including sequences resembling hormone responsive elements T(3)REs and VDREs. We demonstrate that Sat1 promoter driven basal transcription in OK cells was stimulated by tri-iodothyronine. Site-directed mutagenesis identified an imperfect T(3)RE at -454-bp in the Sat1 promoter to be responsible for this activity. This study represents the first characterization of the structure and regulation of the Sat1 gene encoding a SO(4)(2-)/chloride/oxalate anion transporter. PMID- 12590735 TI - Gap junctional intercellular communication in cells isolated from urethane induced tumors in A/J mice. AB - Studies using normal or neoplastically transformed established mouse lung epithelial cell lines revealed a reduction in gap junctional, intercellular communication (GJIC) with transformation. To determine the stage in tumor development at which GJIC is interrupted, we used the well-established model of lung tumors induced in strain A/J mice by urethane. In this system, tumor development follows a well-characterized pattern; hyperplasias, adenomas, and carcinomas are manifested at approximately 8, 16, and 40 weeks after urethane treatment, respectively. GJIC levels were examined using a novel technique where cells are grown on a glass slide, half of which is coated with electrically conductive, optically transparent, indium-tin oxide. An electric pulse that opens transient pores on the plasma membrane is applied in the presence of the fluorescent dye, Lucifer yellow, causing dye penetration into cells growing on the conductive part of the slide. Migration of the dye through gap junctions to nonelectroporated cells growing on the nonconductive area is then microscopically observed under fluorescence illumination. Unexpectedly, primary cells cultured from urethane-induced tumors, even late stage carcinomas, possessed extensive GJIC immediately upon isolation. Upon passage for several months however, these cells lost GJIC. These results suggest that the molecular changes that lead to the formation of the tumor in vivo are not sufficient to interrupt gap junctions. Propagation of tumor cells in culture induces additional alterations that can lead to gap junction closure. PMID- 12590736 TI - Expression of active human blood clotting factor VIII in mammary gland of transgenic rabbits. AB - Human clotting factor VIII is probably the largest protein to be expressed to date in the mammary gland of a transgenic animal, and it requires extensive posttranslational modification to achieve full biological activity. The mammary gland specific construct mWAP-hFVIII-MT-I was injected into the pronuclei of rabbit zygotes, and three transgenic offspring were obtained. Founder 385 showed germ-line transmission of a single integrated copy, and a homozygous line was established from this animal. The rhFVIII was transcribed and translated exclusively in the mammary gland. The activity of rhFVIII in the rabbit milk ranged from 5 to 8% of that found in normal human plasma. Results indicate the suitability of the transgenic rabbit mammary gland for rhFVIII production. PMID- 12590737 TI - The vasa locus in zebrafish: multiple RGG boxes from duplications. AB - Vasa is the most extensively characterized germ cell-specific marker in metazoans. We determined the sequence of the zebrafish vasa locus, which - together with the flanking regions - is 25 kb long. It contains 26 + 1 exons, with an average length of 83 bp. The 5' end of zebrafish vasa is rich in repeats; it includes a peculiar tandem repeat with three units spanning a total of eight exons, each unit containing several tandemly arranged RNA-binding motifs (RGG boxes). The presence of various, nonmicrosatellite type repeats in vasa seems to be universal among the species studied, and could have functional importance during the evolution of the gene, due to the increase in the number of RGG boxes located in these repeats. Analysis of vasa transcripts in zebrafish identified numerous isoforms resulting from alternative splicing (seven) and polyadenylation (four). Mapping to two radiation hybrid panels strengthened the position of the gene on LG10 of zebrafish. PMID- 12590738 TI - The inverted CCAAT motif is an indispensable element of the enhancer B of the mouse major histocompatibility I H2-Kb gene. AB - We have identified a strong binding of nuclear proteins derived from Ltk(-) fibroblasts to the enhancer B of the mouse MHC class I H2-K(b) gene. The inverted CCAAT motif and its adjacent upstream sequences have been revealed as protein binding sites by electrophoretic mobility-shift, methylation interference, and DNase I footprint assays. Specific mutations in the inverted CCAAT motif as well as in the 5'-flanking cytosine pentanucleotide abrogated the formation of the major DNA-protein complex. Transcription of the chloramphenicol acetyltransferase (CAT) reporter gene driven by the H2-K(b) promoter in the Ltk(-) cell line was reduced substantially when a two-nucleotide mutation was introduced into the CCAAT element (CCAATCgcAT). The indicated two-nucleotide mutation decreased transcription initiated from both the homologous and a heterologous promoter. Furthermore, cotransfected MHC class II transactivator (CIITA) elevated the transcription of the reporter gene under the control of the H2-K(b) upstream sequences in the NIH 3T3 cell line. The intact enhancer B involving both the inverted CCAAT motif and the site alpha was found to play an indispensable role in the CIITA-mediated gene transactivation. The band-shift assay with the enhancer B probe revealed forming of a protein complex in a cooperative manner, which was again prevented by mutations in either element. Our results suggest an essential role of the inverted CCAAT element in the constitutive as well as inducible transcription of the mouse MHC class I genes. PMID- 12590740 TI - Simple physical treatment as an effective tool to improve the functional properties of rapeseed (Brassica campestris var. toria) and sesame seed (Sesamum indicum) meals. AB - Dry and 24-h imbibed rapeseeds and sesame seeds were defatted with hexane, and freeze-dried powder was analysed for the functional properties of the meals. The water absorption capacity of the imbibed rapeseed meal and the fat absorption capacity of both the imbibed meals were higher than dry meals. Protein solubility of the rapeseed meal was improved by imbibition and both the imbibed meals exhibited maximum protein solubility at pH 12. The rapeseed meal possessed better foaming properties and viscosity than the sesame seed meal. Imbibition considerably enhanced the foaming properties of the rapeseed meal, while the emulsification properties and viscosity did not change appreciably. Emulsification properties of the sesame meal were higher than those of the rapeseed meal. PMID- 12590739 TI - In vitro accessibility of carotenes from green leafy vegetables cooked with sunflower oil or red palm oil. AB - Although vegetables and fruits are readily available and consumed in most areas of Tanzania, vitamin A deficiency is still prevalent. The objective of the present study was to measure the in vitro accessibility (available for absorption) of alpha-carotene and beta-carotene in vegetable relishes prepared with or without oil. Derived results were used to calculate the contribution of vegetable relish to recommended daily intake of retinol. Five sundried green leafy vegetables from Tanzania were cooked without oil, with sunflower oil or with red palm oil. The total amount and in vitro accessibility of alpha-carotene and beta-carotene from a portion (100 g) of vegetable relish was determined. The in vitro method used simulated the digestion process in the gastrointestinal tract. Carotenoids released after digestion were quantified using high performance liquid chromatography. The total amount of beta-carotene varied between 1211 and 3659 microg/100 g among the five vegetable sources studied. From green leaves cooked without oil, 8-29% of the beta-carotene content became accessible after in vitro digestion and 39-94% from leaves cooked with sunflower oil or red palm oil. Adding red palm oil instead of sunflower oil resulted in about twice as much accessible beta-carotene, due to the high accessibility of its beta-carotene content. The red palm oil contributed also a considerable amount of alpha-carotene. The results showed that by eating vegetable relishes with added oil daily, it should be possible to provide the recommended intake level of vitamin A. PMID- 12590741 TI - Vitamin A deficiency among preschool children in a rural area of Egypt: the results of dietary assessment and biochemical assay. AB - Vitamin A status was evaluated among a cohort of preschool children (mean age 43 months) pertinent to a traditional society in rural Egypt. The Helen Keller International food frequency questionnaire, the 7-day 24-h dietary recall method and serum vitamin A concentrations were the criteria used for the evaluation. Mean values of 280 and 382 retinol equivalents (RE) were the daily estimates of vitamin A intakes among male and female children, respectively. Animal foods made up 39 and 54% of the total vitamin A intake among male and female children, respectively. The aforementioned estimates of total vitamin A intakes were 58 and 81%, respectively, of the FAO/WHO requirements (35 RE/kg). Children from high socio-economic class had significantly higher (P < 0.05) mean vitamin A intake compared with the respective mean intake obtained with poor children. Serum vitamin A concentrations correlated significantly with the respective vitamin A intakes. A 10-week vitamin A intervention trial using either pharmaceutical vitamin A preparations or a food-based strategy consisting of carrot jam led to significant improvement in the growth velocity of the beneficiaries compared with the control group. PMID- 12590742 TI - Brief communication: energy and protein intake in a sample of hospitalized elderly in Hong Kong. AB - The present study investigated energy and protein intake in 17 hospitalized elderly patients in Hong Kong. Food intake was directly measured for breakfast, lunch and dinner over two consecutive days. Energy intakes were low--the meals provided, on average, 57% and 45% of the calculated basal metabolic rate (BMR) and energy requirement (1.27 x BMR), respectively. Protein intake per kilogram of body weight was below 0.75 g in eight subjects. The type of meat served at both lunch and dinner influenced energy and protein intakes. Energy intake was significantly higher when the meals contained pork, and protein intake was significantly higher when chicken was served. The present study highlights the importance of meat in the diet of the hospitalized elderly patient and indicates the influence that this food may have on energy and protein intakes. Effective strategies to increase energy and protein intakes in the hospitalized elderly are urgently required. PMID- 12590743 TI - Changes in alpha-and beta-amylase activities during seed germination of African finger millet. AB - Changes in alpha- and beta-amylase activities in African finger millet (Eleusine coracana (L) Gaertener) were followed during germination. Germination on a small scale was performed at 15 degrees C for 1-10 days and at 20, 25 and 30 degrees C for 1-8 days. alpha- and beta-Amylase activities in malt crude extracts of germinated finger millet were evaluated spectrophotometrically using chromogenic methods. The highest alpha-amylase activity was exhibited in malt flour of finger millet germinated at 15 degrees C for 9 days and at 20 degrees C for 6 days, while the highest beta-amylase activity was displayed in the malt flour germinated for 5 days at 30 degrees C. Thermo-stability of these enzymes in malt extracts was also evaluated. Malt extracts incubated at 40 and 50 degrees C for up to 4 h retained about 84 and 64% of alpha-amylase activities, respectively. There was a substantial decrease in alpha-amylase activity to more than 90% when malt extracts were incubated at 70 and 90 degrees C for 40 and 10 min, respectively. beta-Amylase was completely inactivated when the crude extract was incubated at 70 degrees C for only 10 min. At pH 5.4, alpha-amylase displayed maximum catalytic activity at around 45 degrees C. Optimum temperature for beta amylase activity at pH 6.0 was between 50 and 55 degrees C. Activity staining for alpha-amylase was also performed and three bands of activity were found in malt extract, each possibly representing an isozyme of alpha-amylase from finger millet. PMID- 12590744 TI - Gathering practices of Karen women: questionable contribution to beta-carotene intake. AB - Gathering edible wild plants is widely recognized throughout rural southeast Asia for contributions to micronutrient intakes. Elderly Karen women in two northern Thailand communities (n = 32) were surveyed to determine household gathering practices and use of edible wild plants in the family diet. While all household members gathered most efforts were by women, who collected on average 1.3-3.5 times/week depending on the season. All respondents reported that local availability of wild species with culinary and medicinal value had declined in recent decades. Column chromatography and spectrophotometry were used to determine the beta-carotene content of 22 of the most commonly consumed species. Mean values ranged from 10,290 to 31 microg/100 g sample (858 to 3 microg RE/100 g); 14 species measured at least 2400 microg/100 g (300 microg RE/100 g). Most plants identified with reasonable to high quantities of beta-carotene were eaten raw, steamed for more than 20 min, or were cooked and consumed without the fat source needed to facilitate absorption. The contribution of beta-carotene from these species to Karen diet, therefore, is problematical. PMID- 12590745 TI - Effect of traditional, microwave and industrial cooking on inositol phosphate content in beans, chickpeas and lentils. AB - An high-performance liquid chromatography method for determining inositol phosphate fractions was adapted to legumes. The validity of the method was assessed by estimating the following analytical parameters: linearity (linear response between 125 and 5000 microg inositol hexaphosphate (IP(6))/ml); instrumental precision and method precision (relative standard deviation, %) were 1.9% (IP(6)) for instrumental, and 2.5% (IP(6)) and 8.2% (IP(5)) for method precision. An accuracy was estimated by percentage recovery (72 +/- 3%). The application of this method to raw, conventional, microwave-cooked and ready-to eat beans, chickpeas and lentils gave IP(6) contents ranging from 0.63 g/100 g dry matter in ready-to-eat lentils to 1.87 g/100 g dry matter in raw beans. The IP(6) content was reduced by all the cooking procedures, while the relative percentage of inositol pentaphosphate increased in all the legumes studied, and reached the maximum of 31% (expressed in relation to dry matter) in ready-to-eat beans. PMID- 12590746 TI - Ascorbic acid in diet supplements: loss in the manufacturing process and storage. AB - The ascorbic acid content was determined in 25 different diet supplements commercially available to the consumer in two pharmaceutical forms (pills and ampoules). These products are widely consumed by several population groups (elderly people, sportsmen, adolescents, children, etc.). High-performance liquid chromatography was used as the analytical technique. The proposed method has been validated with good linearity, reproducibility, recovery and accuracy, and can be used in routine analyses and in quality control. The ascorbic acid content in pills ranged from 15.62 to 50.16 mg/g, and in that ampoules from 2.12 to 8.83 mg/ml. Depending on the dosage rates, these levels would represent approximately 20-50% of the daily dietary intake recommended by the National Research Council. Possible losses in the manufacturing process and stability during storage for 30 days at 40 +/- 2 degrees C without light, were tested. In relation to the ascorbic acid concentrations stated on the labels, a loss of 12.0-21.9% in pills and of 11.7-18.0% in ampoules was detected. In relation to the stability conditions, the losses are of 1.8-24.8% in pills and of 10.4-19.3% in ampoules. The pasteurisation and sterilisation processes produced a mean loss of ascorbic acid in ampoules of 2.1 and 1.4%, respectively. A statistically significant direct correlation was observed between ascorbic acid loss and content in proteins, humidity, ash, and fructose. The influence of the pharmaceutical form was also tested. Data revealed that the control of losses during the manufacturing and commercialisation process of these products is necessary to ensure the intake of vitamin C from these products by the consumer. PMID- 12590747 TI - Prevalence of daily breakfast intake, iron deficiency anaemia and awareness of being anaemic among Saudi school students. AB - Iron deficiency anaemia is one of most prevalent nutritional disorders worldwide. It is known to affect the health and cognitive ability of children and adolescents. Studies in Saudi Arabia concentrate only on the population of young children and pregnant females or girls. Studies on the whole school student population is lacking. The objectives of this study were to identify the nutritional habits and the prevalence of anaemia among school students in Jeddah, as well as to recognize the students' awareness of their anaemic nutritional status. Data were collected from a sample of Saudi school children in Jeddah City from 42 boys' and 42 girls' schools during the month of April 2000. Data collection was done by an in-person interview to collect socio-demographic factors, nutritional habits, weight and height. Haemoglobin was measured in a sample of 800 students selected at random from both genders and different age groups. Anaemia was defined according to the new WHO cut-off levels for haemoglobin as: blood haemoglobin <11.5 g/dl for the 5-11 years boys and girls; <12.0 g/dl for 12-14 years boys and girls; <12.0 g/dl for 15+ girls and <13.0 g/dl for 15+ years boys. Proportion and 95% confidence intervals (CI) were calculated and significance was considered when the 95% CI did not overlap. Anaemia was reported among 20.5% of school students. Anaemia was more prevalent among students of at least 12 years as compared to the younger age group. Also, anaemia was more marked among governmental school attendees and those born to low educated mothers. Menstruating girls were at around double the risk of being anaemic than non-menstruating girls. Anaemia was associated with negative impact on school performance and was more marked among those who failed their exams as compared to students with excellent results. Skipping breakfast was reported by 14.9% of students and this habit did not differ by age, sex, body mass index or social class. Skipping breakfast was more marked among students with poor school performance as compared to those with very good or excellent results. Only 34.1% of anaemic school students were aware of being anaemic. Awareness was nearly equal in all age groups and social classes but girls were more aware of their anaemic status than boys. Iron deficiency anaemia appears to be prevalent among school students. At age 12 years and over, low social class and menstruating girls constitute the high-risk groups. Screening is recommended for high-risk groups and school health programs are crucial to improve students' nutritional habits, knowledge and awareness. PMID- 12590748 TI - League tables and school effectiveness: a mathematical model. AB - 'School performance tables', an alphabetical list of secondary schools along with aggregates of their pupils' performances in national tests, have been published in the UK since 1992. Inevitably, the media have responded by publishing ranked 'league tables'. Despite concern over the potentially divisive effect of such tables, the current government has continued to publish this information in the same form. The effect of this information on standards and on the social make-up of the community has been keenly debated. Since there is no control group available that would allow us to investigate this issue directly, we present here a simple mathematical model. Our results indicate that, while random fluctuations from year to year can cause large distortions in the league-table positions, some schools still establish themselves as 'desirable'. To our surprise, we found that 'value-added' tables were no more accurate than tables based on raw exam scores, while a different method of drawing up the tables, in which exam results are averaged over a period of time, appears to give a much more reliable measure of school performance. PMID- 12590749 TI - The construction and analysis of epidemic trees with reference to the 2001 UK foot-and-mouth outbreak. AB - The case-reproduction ratio for the spread of an infectious disease is a critically important concept for understanding dynamics of epidemics and for evaluating impact of control measures on spread of infection. Reliable estimation of this ratio is a problem central to epidemiology and is most often accomplished by fitting dynamic models to data and estimating combinations of parameters that equate to the case-reproduction ratio. Here, we develop a novel parameter-free method that permits direct estimation of the history of transmission events recoverable from detailed observation of a particular epidemic. From these reconstructed 'epidemic trees', case-reproduction ratios can be estimated directly. We develop a bootstrap algorithm that generates percentile intervals for these estimates that shows the procedure to be both precise and robust to possible uncertainties in the historical reconstruction. Identifying and 'pruning' branches from these trees whose occurrence might have been prevented by implementation of more stringent control measures permits estimation of the possible efficacy of these alternative measures. Examination of the cladistic structure of these trees as a function of the distance of each case from its infection source reveals useful insights about the relationship between long distance transmission events and epidemic size. We demonstrate the utility of these methods by applying them to data from the 2001 foot-and-mouth disease outbreak in the UK. PMID- 12590750 TI - Nuclear markers reveal unexpected genetic variation and a Congolese-Nilotic origin of the Lake Victoria cichlid species flock. AB - Phylogenetic analyses based on mitochondrial (mt) DNA have indicated that the cichlid species flock of the Lake Victoria region is derived from a single ancestral species found in East African rivers, closely related to the ancestor of the Lake Malawi cichlid species flock. The Lake Victoria flock contains ten times less mtDNA variation than the Lake Malawi radiation, consistent with current estimates of the ages of the lakes. We present results of a phylogenetic investigation using nuclear (amplified fragment length polymorphism) markers and a wider coverage of riverine haplochromines. We demonstrate that the Lake Victoria-Edward flock is derived from the morphologically and ecologically diverse cichlid genus Thoracochromis from the Congo and Nile, rather than from the phenotypically conservative East African Astatotilapia. This implies that the ability to express much of the morphological diversity found in the species flock may by far pre-date the origin of the flock. Our data indicate that the nuclear diversity of the Lake Victoria-Edward species flock is similar to that of the Lake Malawi flock, indicating that the genetic diversity is considerably older than the 15 000 years that have passed since the lake began to refill. Most of this variation is manifested in trans-species polymorphisms, indicating very recent cladogenesis from a genetically very diverse founder stock. Our data do not confirm strict monophyly of either of the species flocks, but raise the possibility that these flocks have arisen from hybrid swarms. PMID- 12590751 TI - Self-organized lane formation and optimized traffic flow in army ants. AB - We show how the movement rules of individual ants on trails can lead to a collective choice of direction and the formation of distinct traffic lanes that minimize congestion. We develop and evaluate the results of a new model with a quantitative study of the behaviour of the army ant Eciton burchelli. Colonies of this species have up to 200 000 foragers and transport more than 3000 prey items per hour over raiding columns that exceed 100 m. It is an ideal species in which to test the predictions of our model because it forms pheromone trails that are densely populated with very swift ants. The model explores the influences of turning rates and local perception on traffic flow. The behaviour of real army ants is such that they occupy the specific region of parameter space in which lanes form and traffic flow is maximized. PMID- 12590752 TI - Organization of work in the honeybee: a compromise between division of labour and behavioural flexibility. AB - Although the caste concept has been central to our understanding of the organization of work in social insect colonies, the concept has been the subject of considerable recent criticism. Theoretically, it has been suggested that temporal castes are too inflexible to allow a colony to rapidly reallocate labour in response to changing conditions. In addition, several authors have suggested that task switching is so prevalent that it precludes even the possibility of a rigidly controlled temporal caste system. This study addresses these two criticisms by presenting and testing a revision of the temporal caste concept that recognizes two categories of tasks: those that require a physiological specialization for their efficient performance, and those that all workers are equally able to perform. Only those tasks requiring a physiological specialization are relevant to the temporal caste concept. Two castes of honeybees were shown to vary in response to increased nectar influx, which requires a physiological specialization, but not to heat stress, which requires no specialization. This work suggests that the organization of work in social insect colonies reflects a compromise between selection for the benefits of division of labour and opposing selection for flexibility in task allocation. PMID- 12590753 TI - Immune activity elevates energy expenditure of house sparrows: a link between direct and indirect costs? AB - The activation of an immune response is beneficial for organisms but may also have costs that affect fitness. Documented immune costs include those associated with acquisition of special nutrients, as well as immunopathology or autoimmunity. Here, we test whether an experimental induction of the immune system with a non-pathological stimulant can elevate energy turnover in passerine birds. We injected phytohaemagglutinin (PHA), a commonly used mitogen that activates the cell-mediated immune response, into the wing web of house sparrows, Passer domesticus. We then examined energetic costs resulting from this immune activity and related those costs to other physiological activities. We found that PHA injection significantly elevated resting metabolic rate (RMR) of challenged sparrows relative to saline controls. We calculated the total cost of this immune activity to be ca. 4.20 kJ per day (29% RMR), which is equivalent to the cost of production of half of an egg (8.23 kJ egg(-1)) in this species. We suggest that immune activity in wild passerines increases energy expenditure, which in turn may influence important life-history characteristics such as clutch size, timing of breeding or the scheduling of moult. PMID- 12590754 TI - Female age and sperm competition: last-male precedence declines as female age increases. AB - Until very recently, most studies of sperm competition have focused on variation in male competitive ability. However, we now know that a number of reproductive traits, including oviposition rate, use of stored sperm and receptivity to mating, vary with female condition. Because females can play an active part in the movement of sperm within their reproductive tract, sperm competition may be influenced by female condition. Existing studies of sperm competition in fruitflies ignore the effects of female condition, using females that are 3-4 days old and in their reproductive prime. But condition will decline as a female senesces. Here, we examine the effect of female age on the outcome of sperm competition in three strains of the fruitfly, Drosophila melanogaster. Previous studies have shown that female age influences preference for mates and male ejaculation strategies. In this study, we find that when males are mated to females that are older than 17 days, last-male sperm precedence decreases significantly. These results could lead to a greater understanding of the physiological mechanisms that regulate the outcome of sperm competition. PMID- 12590755 TI - A female melanin ornament signals offspring fluctuating asymmetry in the barn owl. AB - Sexual selection theory predicts that males advertise quality by displaying extravagant ornaments. By contrast, whether phenotypic variation in females has a signalling function remains an open question. Here, to our knowledge, we provide the first evidence that a female plumage trait can signal fluctuating asymmetry in the offspring. We experimentally demonstrate in wild barn owls (Tyto alba) that the extent to which females display black spots on their plumage does not only signal offspring parasite resistance as shown in a previous study but also developmental homeostasis in the offspring. A greater number of spotted females produced offspring that had more symmetrical feathers during the period of growth. Males, that pair non-randomly with respect to female plumage spottiness therefore appear to gain substantial benefits by mating with heavily spotted females. Genetic variation in plumage spottiness is nevertheless maintained as the covariation between offspring body mass and mother plumage spottiness varies annually depending on environmental conditions. PMID- 12590756 TI - The Permian mammal-like herbivore Diictodon, the oldest known example of sexually dimorphic armament. AB - Dicynodonts, a highly successful group of Palaeozoic tetrapods, were herbivores with keratinous beaks, and were frequently equipped with large, neomorphic tusks. Diictodon is a particularly abundant dicynodont genus, allowing statistical investigation of its palaeobiology. Anatomical, morphometric and distributional analyses provide evidence of sexual dimorphism, based on the presence or absence of formidable tusks. Tusk occurrence is also correlated with the presence of a cranial boss on the skull roof and, possibly, with greater cranial size. This earliest well-documented example of dimorphic armament suggests that sexual dimorphism, and the complex social behaviour that accompanies it, have long been characteristic of the synapsid lineage. PMID- 12590757 TI - Suppression of skeletal growth in scleractinian corals by decreasing ambient carbonate-ion concentration: a cross-family comparison. AB - Biogenic calcification is influenced by the concentration of available carbonate ions. The recent confirmation of this for hermatypic corals has raised concern over the future of coral reefs because [CO(3)(2-)] is a decreasing function of increasing pCO(2) in the atmosphere. As one of the overriding features of coral reefs is their diversity, understanding the degree of variability between species in their ability to cope with a change in [CO(3)(2-)] is a priority. We cultured four phylogenetically and physiologically different species of hermatypic coral (Acropora verweyi, Galaxea fascicularis, Pavona cactus and Turbinaria reniformis) under 'normal' (280 micromol kg(-1)) and 'low' (140 micromol kg(-1)) carbonate ion concentrations. The effect on skeletogenesis was investigated quantitatively (by calcification rate) and qualitatively (by microstructural appearance of growing crystalline fibres using scanning electron microscopy (SEM)). The 'low carbonate' treatment resulted in a significant suppression of calcification rate and a tendency for weaker crystallization at the distal tips of fibres. However, while the calcification rate was affected uniformly across species (13-18% reduction), the magnitude of the microstructural response was highly species specific: crystallization was most markedly affected in A. verweyi and least in T. reniformis. These results are discussed in relation to past records and future predictions of carbonate variability in the oceans. PMID- 12590758 TI - The spatial resolution of the pinhole eyes of giant clams (Tridacna maxima). AB - Giant clams (Tridacna spp.) have several hundred small pinhole-type eyes on the exposed mantle. They respond by withdrawing the mantle to movements of dark objects, even if these cast no shadow on the animal as a whole. I investigated this 'sight reaction' using black and white square-wave gratings whose phase abruptly changed so that the white areas became dark and vice versa. Gratings with periods of 13.5 degrees were ineffective, but gratings of 20.7 degrees caused partial retraction of mantles or siphons. This implies an acceptance angle for the best-resolving eyes of between 8.7 degrees and 21.8 degrees. A single black spot was effective if its angular diameter was 13.5 degrees but not 11.7 degrees. The mean threshold for the pure dimming of a large field was a decrease of 12.3%, but responses increased in strength up to a dimming of 35%. Anatomically the eyes are ca. 400 microm deep from aperture to receptors, the aperture has a mean diameter of 90 microm and the receptors are 25 microm across. This gives an angular acceptance angle for single receptors of 16.5 degrees, which is completely consistent with the behavioural measurements. PMID- 12590759 TI - Sub-lethal effects of pathogens can lead to the evolution of lower virulence in multiple infections. AB - According to current evolutionary dogma, multiple infections generally increase a parasite's virulence (i.e. reduce the host's reproductive success). The basic idea is that the competitive interactions among strains of parasites developing within a single host select individual parasites to exploit their host more rapidly than their competitors (thereby causing an increase in virulence) to ensure their transmission. Although experimental evidence is scarce, it often contradicts the theoretical expectation by suggesting that multiple infections lead to decreased virulence. Here, we present a theoretical model to explain this contradiction and show that the evolutionary outcome of multiple infections depends on the characteristics of the interaction between the host and its parasite. If we assume, as current models do, that parasites have only lethal effects on their host, multiple infections indeed increase virulence. By contrast, if parasites have sub-lethal effects on their host (such as reduced growth) and, in particular, if these effects feed back onto the parasites to reduce their rate of development, then multiplicity of infection generally leads to lower virulence. PMID- 12590760 TI - The flowerpiercers' hook: an experimental test of an evolutionary trade-off. AB - The evolution of features that enhance an organism's performance in one activity can adversely affect its performance in another. We used an experimental approach to document a trade-off associated with the evolution of the long hook at the tip of the bill of birds belonging to the genus Diglossa (flowerpiercers). In Diglossa, the more derived flower-robbing nectarivorous species have maxillae (upper jaws) that terminate in enlarged curved hooks. The ancestral frugivorous species have maxillae with relatively small hooks. We mimicked bill evolution by clipping the terminal bill hook of nectarivorous Cinnamon-bellied Flowerpiercers (Diglossa baritula) to resemble the frugivorous condition. We found that birds with experimentally shortened bills ingested fruit more efficiently, but had a reduced ability to rob flowers. Birds with intact bills, by contrast, were good flower robbers but poor frugivores. The evolution of a hooked bill endowed flowerpiercers with the ability to efficiently pierce flowers and extract nectar, but hindered their efficiency to feed on fruit. PMID- 12590761 TI - From reciprocity to unconditional altruism through signalling benefits. AB - Cooperation among genetically unrelated individuals is commonly explained by the potential for future reciprocity or by the risk of being punished by group members. However, unconditional altruism is more difficult to explain. We demonstrate that unconditional altruism can evolve as a costly signal of individual quality (i.e. a handicap) as a consequence of reciprocal altruism. This is because the emergent correlation between altruism and individual quality in reciprocity games can facilitate the use of altruism as a quality indicator in a much wider context, outside the reciprocity game, thus affecting its further evolution through signalling benefits. Our model, based on multitype evolutionary game theory shows that, when the additive signalling benefit of donating help exceeds the cost for only some individuals (of high-quality state) but not for others (of low-quality state), the population possesses an evolutionarily stable strategy (ESS) profile wherein high-quality individuals cooperate unconditionally while low-quality individuals defect or play tit-for-tat (TfT). Hence, as predicted by Zahavi's handicap model, signalling benefits of altruistic acts can establish a stable generosity by high-quality individuals that no longer depends on the probability of future reciprocation or punishment. PMID- 12590762 TI - Adaptation of metabolism and evaporative water loss along an aridity gradient. AB - Broad-scale comparisons of birds indicate the possibility of adaptive modification of basal metabolic rate (BMR) and total evaporative water loss (TEWL) in species from desert environments, but these might be confounded by phylogeny or phenotypic plasticity. This study relates variation in avian BMR and TEWL to a continuously varying measure of environment, aridity. We test the hypotheses that BMR and TEWL are reduced along an aridity gradient within the lark family (Alaudidae), and investigate the role of phylogenetic inertia. For 12 species of lark, BMR and TEWL decreased along a gradient of increasing aridity, a finding consistent with our proposals. We constructed a phylogeny for 22 species of lark based on sequences of two mitochondrial genes, and investigated whether phylogenetic affinity played a part in the correlation of phenotype and environment. A test for serial independence of the data for mass-corrected TEWL and aridity showed no influence of phylogeny on our findings. However, we did discover a significant phylogenetic effect in mass-corrected data for BMR, a result attributable to common phylogenetic history or to common ecological factors. A test of the relationship between BMR and aridity using phylogenetic independent constrasts was consistent with our previous analysis: BMR decreased with increasing aridity. PMID- 12590763 TI - The phenomenology of niche evolution via quantitative traits in a 'black-hole' sink. AB - Previous studies of adaptive evolution in sink habitats (in which isolated populations of a species cannot persist deterministically) have highlighted the importance of demographic constraints in slowing such evolution, and of immigration in facilitating adaptation. These studies have relied upon either single-locus models or deterministic quantitative genetic formulations. We use individual-based simulations to examine adaptive evolution in a 'black-hole' sink environment where fitness is governed by a polygenic character. The simulations track both the number of individuals and their multi-locus genotypes, and incorporate, in a natural manner, both demographic and genetic stochastic processes. In agreement with previous studies, our findings reveal the central parts played by demographic constraints and immigration in adaptation within a sink (adaptation is more difficult in environments with low absolute fitness, and higher immigration can accelerate adaptation). A novel finding is that there is a 'punctuational' pattern in adaptive evolution in sink environments. Populations typically stay maladapted for a long time, and then rapidly shift into a relatively adapted state, in which persistence no longer depends upon recurrent immigration. PMID- 12590765 TI - Daughters on request: about helpers and egg sexes in the Seychelles warbler. AB - The Seychelles warbler (Acrocephalus sechellensis) was an endangered endemic of the Seychelles islands where, until 1988, the entire population of ca. 320 birds was restricted to the one island of Cousin Island (29 ha). Additional breeding populations were successfully established on the islands of Aride (68 ha, 1988) and Cousine (26 ha, 1990) and now with the existence of ca. 2000 warblers on three islands the conservation status of the warbler has improved from endangered to vulnerable. Emigration from the island is extremely rare, so birds that disappeared were known to have died. Almost every bird on Cousin Island has been individually colour ringed and monitored throughout all breeding attempts during a 17-year period (1985-2002; total ca. 2400 birds). These birds were also blood sampled for molecular parentage and sex analyses. Therefore the lifetime reproductive success of many birds is known. Although warblers can breed independently in their first year, some individuals remain in their natal territory as subordinates, and often help by providing nourishment to non descendant offspring. The frequency of 'helping' is affected by habitat saturation and variation in territory quality (insect prey availability). The long-term benefits of helping are higher for daughters than for sons, and it is therefore no wonder that most helpers are daughters from previous broods. Furthermore, on low-quality territories breeding pairs raising sons gain higher fitness benefits than by raising daughters, and vice versa on high-quality territories. Female breeders adaptively modify the sex of their single-egg clutches according to territory quality: male eggs on low quality and female eggs on high quality. The Seychelles warbler is a beautiful example of behavioural and life-history adaptations to restricted circumstances. PMID- 12590766 TI - Ecological stress and sex evolution in soil microfungi. AB - The elucidation of the origin and maintenance of sex is a major unsolved problem in evolutionary biology. A number of hypotheses have been elaborated, but the scarcity of empirical data limits further progress. During recent years, the general inclination has changed towards pluralistic models of sex evolution, due partly to an increased diversity of studied organisms. Fungi are among the most promising organisms for testing sexual causation, as demonstrated in recent laboratory experiments. However, reconciling theory and evidence necessitates critical field observations. Here, we report new estimates of the distribution of morphologically sexual and asexual soil microfungi in nature, which indicate a remarkable trend towards increased sexuality with increasing climatic stress. PMID- 12590767 TI - The maintenance of sex in parasites. AB - The maintenance of sex is an unresolved paradox in evolutionary biology, given the inherent twofold fitness advantage for asexuals. Parasitic helminths offer a unique opportunity to address this enigma. Parasites that can create novel antigenic strains are able to escape pre-existing host immunity. Viruses produce diversity through mutation with rapid clonal proliferation. The long generation times of helminth parasites prevent them from adopting this strategy. Instead, we argue that sexual reproduction enables parasitic helminths to rapidly generate strain diversity. We use both a stochastic, individual-based model and a simple analytical model to assess the selective value of sexual versus asexual reproduction in helminth parasites. We demonstrate that sexual reproduction can more easily produce and maintain strain diversity than asexual reproduction for long-lived parasites. We also show that sexual parasite populations are resistant to invasion by rare asexual mutants. These results are robust to high levels of cross-immunity between strains. We suggest that the enhancement of strain diversity, despite stochastic extinction of strains, may be critical to the evolutionary success of sex in long-lived parasites. PMID- 12590768 TI - The origin and maintenance of nuclear endosperms: viewing development through a phylogenetic lens. AB - The endosperm develops in fertilized ovules of angiosperms following fertilization of the central cell and nuclei in the female gametophyte. Endosperms differ in whether, and which, nuclear divisions are followed by cellular divisions; the variants are classified as cellular, nuclear or helobial. Functional correlates of this variation are little understood. Phylogenetic methods provide a powerful means of exploring taxonomic variation and phylogenetic patterns, to frame questions regarding biological processes. Data on endosperms across angiosperms were analysed in a phylogenetic context in order to determine homologies and detect biases in the direction of evolutionary transitions. Analyses confirm that neither all nuclear nor all helobial endosperms are homologous, raise the possibility that cellular development is a reversal in some derived angiosperms (e.g. asterids) and show that a statistically significant bias towards evolution of nuclear endosperms (and against reversals) prevails in angiosperms as a whole. This bias suggests strong selective advantages to having nuclear endosperm, developmental constraints to reversals or both. Homologies suggest that the microtubular cycle and cellularization pattern characteristic of reproductive cells across land plants may have been independently co-opted during multiple origins of nuclear endosperms, but information on cellular endosperms is essential to investigate further. PMID- 12590769 TI - Cooperation, virulence and siderophore production in bacterial parasites. AB - Kin selection theory predicts that the damage to a host resulting from parasite infection (parasite virulence) will be negatively correlated to the relatedness between parasites within the host. This occurs because a lower relatedness leads to greater competition for host resources, which favours rapid growth to achieve greater relative success within the host, and that higher parasite growth rate leads to higher virulence. We show that a biological feature of bacterial infections can lead to the opposite prediction: a positive correlation between relatedness and virulence. This occurs because a high relatedness can favour greater (cooperative) production of molecules that scavenge iron (siderophores), which results in higher growth rates and virulence. More generally, the same underlying idea can predict a positive relationship between relatedness and virulence in any case where parasites can cooperate to increase their growth rate; other examples include immune suppression and the production of biofilms to aid colonization. PMID- 12590770 TI - Realized heritability of personalities in the great tit (Parus major). AB - Behaviour under conditions of mild stress shows consistent patterns in all vertebrates: exploratory behaviour, boldness, aggressiveness covary in the same way. The existence of highly consistent individual variation in these behavioural strategies, also referred to as personalities or coping styles, allows us to measure the behaviour under standardized conditions on birds bred in captivity, link the standardized measurements to the behaviour under natural conditions and measure natural selection in the field. We have bred the great tit (Parus major), a classical model species for the study of behaviour under natural conditions, in captivity. Here, we report a realized heritability of 54 +/- 5% for early exploratory behaviour, based on four generations of bi-directional artificial selection. In addition to this, we measured hand-reared juveniles and their wild caught parents in the laboratory. The heritability found in the mid-offspring-mid parent regression was significantly different from zero. We have thus established the presence of considerable amounts of genetic variation for personality types in a wild bird. PMID- 12590771 TI - Sex chromosome evolution and speciation in Ficedula flycatchers. AB - Speciation is the combination of evolutionary processes that leads to the reproductive isolation of different populations. We investigate the significance of sex-chromosome evolution on the development of post- and prezygotic isolation in two naturally hybridizing Ficedula flycatcher species. Applying a tag-array based mini-sequencing assay to genotype single nucleotide polymorphisms (SNPs) and interspecific substitutions, we demonstrate rather extensive hybridization and backcrossing in sympatry. However, gene flow across the partial postzygotic barrier (introgression) is almost exclusively restricted to autosomal loci, suggesting strong selection against introgression of sex-linked genes. In addition to this partial postzygotic barrier, character displacement of male plumage characteristics has previously been shown to reinforce prezygotic isolation in these birds. We show that male plumage traits involved in reinforcing prezygotic isolation are sex linked. These results suggest a major role of sex-chromosome evolution in mediating post- and prezygotic barriers to gene flow and point to a causal link in the development of the two forms of reproductive isolation. PMID- 12590772 TI - Ultraviolet signals in birds are special. AB - Recent behavioural experiments have shown that birds use ultraviolet (UV) reflective and fluorescent plumage as cues in mate choice. It remains controversial, however, whether such UV signals play a special role in sexual communication, or whether they are part of general plumage coloration. We use a comparative approach to test for a general association between sexual signalling and either UV-reflective or fluorescent plumage. Among the species surveyed, 72% have UV colours and there is a significant positive association between UV reflectance and courtship displays. Among parrots (Psittaciformes), 68% of surveyed species have fluorescent plumage, and again there is a strong positive association between courtship displays and fluorescence. These associations are not artefacts of the plumage used in courtship displays, being generally more 'colourful' because there is no association between display and colours lacking UV reflectance or fluorescence. Equally, these associations are not phylogenetic artefacts because all results remain unchanged when families or genera, rather than species, are used as independent data points. We also find that, in parrots, fluorescent plumage is usually found adjacent to UV-reflective plumage. Using a simple visual model to examine one parrot, the budgerigar Melopsittacus undulatus, we show that the juxtaposition of UV-reflective and fluorescent plumage leads to a 25-fold increase in chromatic contrast to the budgerigar's visual system. Taken together, these results suggest that signals based on UV contrast are of special importance in the context of active sexual displays. We review briefly six hypotheses on why this may be the case: suitability for short range signalling; high contrast with backgrounds; invisibility to predators; exploitation of pre-existing sensory biases; advertisement of feather structure; and amplification of behavioural signals. PMID- 12590773 TI - Common language or Tower of Babel? On the evolutionary dynamics of signals and their meanings. AB - We investigate how the evolution of communication strategies affects signal credibility when there is common interest as well as a conflict between communicating individuals. Taking alarm calls as an example, we show that if the temptation to cheat is low, a single signal is used in the population. If the temptation increases cheaters will erode the credibility of a signal, and an honest mutant using a different signal ('a private code') will be very successful until this, in turn, is cracked by cheaters. In such a system, signal use fluctuates in time and space and hence the meaning of a given signal is not constant. When the temptation to cheat is too large, no honest communication can maintain itself in a Tower of Babel of many signals. We discuss our analysis in the light of the Green Beard mechanism for the evolution of altruism. PMID- 12590774 TI - Copulatory courtship signals male genetic quality in cucumber beetles. AB - In the spotted cucumber beetle, Diabrotica undecimpunctata howardi (Coleoptera: Chrysomelidae), males court females during copulation by stroking them with their antennae. Stroking occurs exclusively during the first stages of copulation, after a male has penetrated a female's vaginal duct but before he is allowed access to her bursa copulatrix. Females accept the spermatophore of fast-stroking males and reject those of slow-stroking males by relaxing or constricting muscles distorting the vaginal duct. Here, we measure the repeatability of stroking behaviour within males, examine the effect of losing one antenna on male attractiveness and test whether such female control results in direct phenotypic benefits for the discriminating female or indirect genetic benefits that appear in her offspring. We also use a half-sibling design to quantify the variance and heritability of stroking speed and endurance. Female beetles were paired with a male that was known to stroke either quickly or slowly. No difference was found in the resulting fecundity or egg-hatching rate of the females, or in the survivorship, development rate, size, age at first reproduction or fecundity of their offspring indicating that no direct benefits are gained by discriminating among males on the basis of stroking speed. There were, however, good-genes benefits for the mates of fast-stroking males. Offspring of fast-stroking fathers were also fast strokers and were more likely to be accepted as mates than offspring of slow-stroking fathers. There was substantial variance among sires in stroking speed and endurance and the heritability of each trait was high. The antennal stroking rate was highly repeatable in successive mating attempts and males with only one antenna were not accepted as mates. The repeatability within males, variability between males and heritability between generations of copulatory stroking combine to provide females with a reliable and honest signal of the genetic quality of courting males. PMID- 12590775 TI - Is neural filling-in necessary to explain the perceptual completion of motion and depth information? AB - Retinal activity is the first stage of visual perception. Retinal sampling is non uniform and not continuous, yet visual experience is not characterized by holes and discontinuities in the world. How does the brain achieve this perceptual completion? Fifty years ago, it was suggested that visual perception involves a two-stage process of (i) edge detection followed by (ii) neural filling-in of surface properties. We examine whether this general hypothesis can account for the specific example of perceptual completion of a small target surrounded by dynamic dots (an 'artificial scotoma'), a phenomenon argued to provide insight into the mechanisms responsible for perception. We degrade the target's borders using first blur and then depth continuity, and find that border degradation does not influence time to target disappearance. This indicates that important information for the continuity of target perception is conveyed at a coarse spatial scale. We suggest that target disappearance could result from adaptation that is not specific to borders, and question the need to hypothesize an active filling-in process to explain this phenomenon. PMID- 12590776 TI - Rescue of a severely bottlenecked wolf (Canis lupus) population by a single immigrant. AB - The fragmentation of populations is an increasingly important problem in the conservation of endangered species. Under these conditions, rare migration events may have important effects for the rescue of small and inbred populations. However, the relevance of such migration events to genetically depauperate natural populations is not supported by empirical data. We show here that the genetic diversity of the severely bottlenecked and geographically isolated Scandinavian population of grey wolves (Canis lupus), founded by only two individuals, was recovered by the arrival of a single immigrant. Before the arrival of this immigrant, for several generations the population comprised only a single breeding pack, necessarily involving matings between close relatives and resulting in a subsequent decline in individual heterozygosity. With the arrival of just a single immigrant, there is evidence of increased heterozygosity, significant outbreeding (inbreeding avoidance), a rapid spread of new alleles and exponential population growth. Our results imply that even rare interpopulation migration can lead to the rescue and recovery of isolated and endangered natural populations. PMID- 12590777 TI - Modelling pathogen transmission: the interrelationship between local and global approaches. AB - We describe two spatial (cellular automaton) host-pathogen models with contrasting types of transmission, where the biologically realistic transmission mechanisms are based entirely on 'local' interactions. The two models, fixed contact area (FCA) and fixed contact number (FCN), may be viewed as local 'equivalents' of commonly used global density- (and frequency-) dependent models. Their outputs are compared with each other and with the patterns generated by these global terms. In the FCN model, unoccupied cells are bypassed, but in the FCA model these impede pathogen spread, extending the period of the epidemic and reducing the prevalence of infection when the pathogen persists. Crucially, generalized linear modelling reveals that the global transmission terms betaSI and beta'SI/N are equally good at describing transmission in both the FCA and FCN models when infected individuals are homogeneously distributed and N is approximately constant, as at the quasi-equilibrium. However, when N varies, the global frequency-dependent term beta'SI/N is better than the density-dependent one, betaSI, at describing transmission in both the FCA and FCN models. Our approach may be used more generally to compare different local contact structures and select the most appropriate global transmission term. PMID- 12590778 TI - Impact of asbestos concentrations in floor tiles on exposure during removal. AB - This study investigated personal airborne exposure to asbestos during its abatement in two projects that had floor tile with different concentrations of asbestos. Abatement of floor tile with the higher percent asbestos exhibited the highest personal exposure level. Airborne exposure concentrations of asbestos between these abatement projects were found to be statistically significant. However, since neither had levels above occupational exposure standards this difference was not considered important, a type two error (false positive). It is suggested that when establishing negative exposure assessments, priority be given to process/work practices, with percent of contaminant in the building component being of less importance. Time period of applicability of data from negative exposure assessments to future projects should be considered indefinite. Neither within- or between-worker variation was predominant, in this study. Likelihood of exceeding occupational exposure standards during abatement of asbestos-containing floor tile, even for materials containing up to 15% asbestos, is low. Regulation of floor tile as a special category or group of asbestos-containing material appears to be warranted. Current schemes in the United States that regulate floor tile in the same group as other asbestos-containing materials would appear to be a legislating of science. PMID- 12590779 TI - Lead-poisoned wildfowl in Spain: a significant threat for human consumers. AB - Each year, 1.2 million Spanish hunters and shooters discharge 6,000 t of lead shot, of which 30-50 t are deposited in wetlands of this European country. Waterfowl may accidentally ingest lead pellets in these aquatic habitats and become fatally lead poisoned. It has been estimated that 50,000 birds die from this cause in Spain each year, but many more are chronically affected. Most of them are species that can be hunted legally, and the lead toxicosis enhances their susceptibility to being killed by hunting. Consequently, about 30,000 waterfowl hunters and their families, especially children, are at risk from secondary lead ingestion from these poisoned birds. The consumption of a single liver (often eaten in Spain) from any waterfowl shot in this country may result in the direct uptake of 0.01-2.3 mg of lead in 40.4% of cases. This is based on the percentage of 411 analyzed waterfowl having liver lead contents over 0.5 mg kg(-1) wet weight, the maximum lead level in poultry offal that current EU regulations permit. Therefore, health management authorities should draw urgent attention to this environmental problem that presents such an established risk to human health. PMID- 12590780 TI - A comparison of traditional and recently developed methods for monitoring surface hygiene within the food industry: an industry trial. AB - A rapid, instrument-free, hygiene monitoring system, based on protein detection, was assessed as a means to evaluate the cleanliness of food contact surfaces within four different food processing environments. Its performance was compared to that of both ATP bioluminescence and a traditional agar-based microbiological method. Each surface was sampled using all three hygiene monitoring systems both before and after each of the production plants had carried out their normal cleaning procedures. In both cases, there was a significant difference (p < 0.05) between the number of surfaces that were passed or failed using each of the tests. In general the number of surfaces that were deemed acceptable for food production increased after cleaning but the level of agreement between test methods differed depending on the type of production facility sampled. Protein detection was most likely to fail surfaces within the baking facility, whereas ATP bioluminescence and traditional microbiology were most likely to fail surfaces within a frozen ready-meal and a cheese production unit respectively. The implications of these results are discussed in relation to plant trials of hygiene monitoring systems, the cleaning process itself, failures in disinfection, as well as the need for a combined methodological approach for monitoring cleanliness. PMID- 12590781 TI - Balancing risks in the management of contaminated first nations fisheries. AB - In the 1980s and 1990s, the Government of Canada closed and/or issued advisories for a number of shellfish fisheries in coastal areas of British Columbia because of dioxin contamination. Only the direct health risks (i.e., cancer) of consuming contaminated shellfish for the general population were considered by the Government in the formulation of risk management options. A focus on the direct risks does not provide an adequate basis for risk decisions as the countervailing risks which may be created from management measures may easily be overlooked. This study describes the potential health impacts of risk management options for aboriginal coastal peoples in the management of dioxin contamination. Gold River and Powell River in British Columbia, Canada, are the areas of focus. The cancer risks of consuming dioxin contaminated shellfish for these sites are estimated. To assess the countervailing risks of management decisions for comparison, a scenario was developed in which First Nations peoples substitute shellfish with store-bought foods in their diets in the event of a fishery closure or advisory. Increases in mortality due to coronary heart disease are estimated. The results suggest that the health risks of dietary changes among aboriginal peoples may be as significant as those related to eating dioxin contaminated shellfish. PMID- 12590782 TI - Daily consumption of wild fish and game: exposures of high end recreationists. AB - There has been widespread interest in the exposure of indigenous and subsistence populations to a variety of contaminants through the consumption of wild fish and game, yet there is little information on recreationists. Information on wild fish and game consumption in South Carolina are presented here, data on other sources of animal protein are presented elsewhere. Data are presented for use in probabilistic risk assessment. Fish and deer were the most commonly eaten wild caught foods. High-end consumers were eating at nearly 10x the median consumption rate for most wild-caught foods. There were significant differences in consumption as a function of age and ethnicity that should be taken into account when conducting risk assessments for the consumption of wild-caught meat and fish. Men consumed more of these foods, while white sportsmen consumed more deer and black sportsmen consumed more fish. Similar data are not generally available, either for the U.S. as a whole, or for specific geographical locations, although there are data for self-caught fish because of the issuance of consumption advisories for U.S. waters. PMID- 12590783 TI - The rate of gastroenteritis in a large city before and after chlorination. AB - There is conflicting evidence about the contribution of drinking water to endemic community gastroenteritis in water supplies which meet conventional microbiological standards with some studies reporting associations between drinking water and endemic disease and others finding no evidence that water is implicated in disease. This study reports the results of an ecological study investigating the effect on community gastroenteritis of chlorinating a city of over 3 million people in the mid 1970s. Prior to chlorination faecal coliforms were regularly identified in the water. Admissions for gastroenteritis and attendances to the Emergency Department of the Royal Children's Hospital, Melbourne's major children's hospital, were measured between 1974 and 1980 inclusive and the influence of chlorination on rates of gastroenteritis was examined. No statistically significant difference was found in the number of admissions or emergency department visits before and after chlorination of the water supply. The study highlights the need for caution when interpreting the relationships between drinking water and gastroenteritis. The result suggests that water was not a dominant contributor to the burden of gastrointestinal disease in the community despite faecal coliforms being present in the water supply. It indicates the need for caution when attributing significant illness to drinking water when there have been only small changes in water quality without first stringently reviewing the studies methodology and understanding their limitations. PMID- 12590784 TI - Highly active antiretroviral treatment in countries with very limited resources: do we have cheaper alternatives? PMID- 12590785 TI - Relation between Gram-stain and clinical criteria for diagnosing bacterial vaginosis with special reference to Gram grade II evaluation. AB - The aim of this study was to analyse how the results of Gram-staining vaginal smears correlated with the clinical criteria for determining the existence of bacterial vaginosis (BV) and, in particular, how the category defined as 'intermediate' or Gram grade II did so and its significance. Women attending an antenatal clinic with an abnormal vaginal flora, that is those who had Gram stains of grades II or III, the latter considered to equate with BV, were given clindamycin or a placebo intravaginally and examined again on up to three occasions. Gram-stain readings of grade III correlated with the clinical criteria for BV on 356 (91.7%) of 388 occasions. Grade II readings covered the spectrum of clinical criteria and correlated with those for BV on 35 (37.2%) of 94 occasions. Grade I, recorded 231 times and seen usually after clindamycin treatment, was associated with BV only once. The sensitivity, specificity, positive predictive value and negative predictive value of the Gram stain for the diagnosis of BV, based on a combination of grades II and III, were 99.7%, 71.6%, 81% and 99.6%, respectively; based on grade III only, the values were 99.7%, 87.7%, 91.6% and 99.6%, respectively. Women reported a malodorous vaginal discharge on 49.2% of the occasions a grade III flora was seen and 13.3% of the times grade II was recorded. It was not associated with grade I and would seem a useful adjunct to the accepted clinical criteria for diagnosing BV. Each of the clinical criteria was found in about equal proportions (87%-91%) for women whose Gram grade was III. For grade II, an increased discharge was noted most often (76.5%) and 'clue' cells least often (24.5%). A positive amine test was the most specific, being associated with <1% of grade I smears. Of women with grade III status, 91% reverted to grade I after treatment with clindamycin for three days. In contrast, of women with grade II status, 53% reverted to grade I, as did 47% of those who were given a placebo. The 'intermediate' (grade II) category is a Gram-stain diagnosis and not one that can be made clinically. It is important to recognize as a distinct entity not only because amalgamation with grade III diminishes the specificity and positive predictive value of the Gram-stain for diagnosing BV, but also because women of grade II status usually fail to respond to clindamycin treatment, whereas those of grade III do not. PMID- 12590786 TI - Effectiveness, satisfaction and compliance with imiquimod in the treatment of external anogenital warts. AB - This study aims to evaluate the effectiveness of imiquimod treatment of external anogenital warts and analyse its possible relationship with patient treatment satisfaction and compliance under conditions of routine clinical practice. An observational, prospective and multi-centre study was conducted in a cohort of 559 patients with external anogenital warts. Imiquimod 5% cream was administered three times a week until the end of treatment (complete wart clearance or up to a maximum of 16 weeks). Effectiveness and compliance were evaluated at four weeks and again at the end of treatment, when satisfaction was also assessed. Complete wart clearance was experienced by 66.6% of patients at the end of treatment and a 50% or greater reduction in total wart area occurred in 79.5%. Imiquimod was more effective in patients who were more satisfied and compliant with treatment. Under conditions of routine clinical practice, imiquimod is an effective treatment for external anogenital warts. PMID- 12590787 TI - 'I'm less at risk than most guys': gay men's unrealistic optimism about becoming infected with HIV. AB - Among the self-justifications that gay men use when engaging in high-risk sex is the thought that they are less at risk than most gay men. Two explanatory models of such 'unrealistic optimism' (UO) have been proposed: while the motivational account holds that UO arises because it serves the function of bringing comfort, the cognitive account holds that UO serves no particular function, being simply a by-product of normal cognitive strategies. This study investigated predictions derived from the motivational account. Gay men uninfected with HIV (n = 88) answered two test questions, requiring them to estimate, respectively, their own risk of becoming infected and that of the average gay man. The questions were presented in the two possible orders, and were either separated or not separated by unrelated filler material. The great majority of the men (89%) exhibited UO. Neither question order nor the interpolation of filler material affected responses to either test question. The results were inconsistent with the motivational account, but explicable in terms of the cognitive account. It seems that the cognitive account provides the better explanation of at least that form of UO measured in this study. Implications for AIDS educators are discussed. PMID- 12590788 TI - HIV-related testicular cancer. PMID- 12590789 TI - 'Shelf life' of Trichomonas vaginalis. AB - The diagnosis of Trichomonas vaginalis (TV) infection in women may be made by observing motile trichomonads in wet-mount preparations of vaginal discharge. The duration for which these organisms can be identified in such samples is unknown. We sought to assess this by performing wet-mount microscopy on samples from female patients immediately, and then in positive samples, every 10 minutes until motile trichomonads could no longer be identified. Of the 65 wet-mount specimens positive for TV, a cumulative total of 13 (20%) samples had become negative by 10 minutes, 23 (35%) samples by 30 minutes and 51 samples (78%) by two hours, with the remainder exceeding two hours. We conclude that one-fifth of wet-mount vaginal preparations initially positive for motile TV become negative within 10 minutes of the initial, immediate reading. In order to maximize the sensitivity of this widely used test it is recommended that all specimens be examined immediately after they are taken. PMID- 12590790 TI - Predictors of seropositivity to herpes simplex virus type 2 in women. AB - Five hundred and twenty consecutive women newly attending a genitourinary medicine clinic who participated in a study of sexual behaviour were also tested for type-specific antibody to herpes simplex virus type 2; 135 (26%) were seropositive, of whom only 29 (21.5%) had had clinical evidence of genital herpes. Seropositive women were much more likely to have a past history of genital herpes (odds ratio [OR] 173). They were also more likely to be black non UK born (OR 14), aged 30 years or over (OR 6), to have had 6-20 sexual partners (OR 3-4), especially from abroad (OR 12), to be unemployed (OR 6) or blue collar workers (OR 4), to have smoked cigarettes (OR 2) and to have practised peno-anal penetration (OR 5). Disease predictors included a past history of pelvic inflammatory disease (OR 63) and bacterial vaginosis (OR 3). Unexpected predictors were only one sexual partner (OR 5) and no non-regular partners (OR 5). Commencing intercourse before 16 years of age showed a protective effect (OR 0.2) and so did use of oral contraception (OR 0.5). Our findings show that infection with HSV-2 is associated with a wider range of morbidity and also emphasize the role of male sexual partner selection in the transmission of infection. PMID- 12590791 TI - Sensitivity and specificity of rapid HIV testing of pregnant women in India. AB - OBJECTIVE: Efforts to prevent HIV transmission from mother to infants in settings like India may benefit from the availability of reliable methods for rapid and simple HIV screening. Data from India on the reliability of rapid HIV test kits are limited and there are no data on the use of rapid HIV tests for screening of pregnant women. METHODS: Pregnant women attending an antenatal clinic and delivery room in Pune agreed to participate in an evaluation of five rapid HIV tests, including (a) a saliva brush test (Oraquick HIV-1/2, Orasure Technologies Inc.), (b) a rapid plasma test (Oraquick HIV-1/2) and (c) three rapid finger prick tests (Oraquick HIV-1/2; HIV-1/2 Determine, Abbott; NEVA HIV-1/2 Cadila). Results of the rapid tests were compared with three commercial plasma enzyme immunoassay (EIA) tests (Innotest HIV AB EIA, Lab systems/ELISCAN HIV AB EIA, UBI HIV Ab EIA). RESULTS: Between September 2000 and October 1, 2001, 1258 pregnant women were screened for HIV using these rapid tests. Forty-four (3.49%) of the specimens were HIV-antibody-positive by at least two plasma EIA tests. All of the rapid HIV tests demonstrated excellent specificity (96-100%). The sensitivity of the rapid tests ranged from 75-94%. The combined sensitivity and specificity of a two-step algorithm for rapid HIV testing was excellent for a number of combinations of the five rapid finger stick tests. CONCLUSION: In this relatively low HIV prevalence population of pregnant women in India, the sensitivity of the rapid HIV tests varied, when compared to a dual EIA algorithm. In general, the specificity of all the rapid tests was excellent, with very few false positive HIV tests. Based upon these data, two different rapid HIV tests for screening pregnant women in India would be highly sensitive, with excellent specificity to reliably prevent inappropriate use of antiretroviral therapy for prevention of vertical HIV transmission. PMID- 12590792 TI - Lack of ageing cohort effect among sexually acquired HIV infections in Hong Kong. AB - We studied HIV rate in different age populations and whether there has been on going new infections in Hong Kong, by examining the age characteristics and their temporal trend of reported HIV infections. It was found that people in the age group 25-34 were worst hit by the local epidemic. Ageing cohort effect was not apparent for the age pattern of sexually-acquired infections reported from 1987 to 2000, with the annual median ages ranged from 31.5 to 36 years (regression coefficient 0.14, 95% confidence interval -0.14 to 0.42, P = 0.295). Similar temporal age patterns were observed for individual subgroups of homo-/bisexuals, heterosexuals and female heterosexuals. The findings suggested the occurrence of new HIV infections over the years in Hong Kong, despite maintenance of a low prevalence. Albeit capturing largely prevalent infections, reported HIV/AIDS data could be tapped for understanding the evolving epidemic pattern to better evaluate and target current prevention programmes. PMID- 12590793 TI - Seroprevalence of sexually transmitted viruses in Korean populations including HIV-seropositive individuals. AB - In order to investigate the prevalence of sexually transmitted viruses (STVs) in Korean populations, the prevalence of specific antibodies to herpes simplex type 2 (HSV-2) and hepatitis C virus (HCV) and surface antigen to hepatitis B virus (HBsAg) was evaluated in blood donors (n = 200), voluntary visitors for STI testing in public health centre (n = 204), HIV seropositive individuals (n = 200) and commercial sex workers (CSWs) (n = 200). All blood samples were tested using commercial enzyme-linked immunosorbent assay (ELISA) kits that detect type specific IgG to HSV-2 and HBsAg and anti-HCV. The prevalence of type-specific IgG to HSV-2 among the above four groups was 5.0%, 21.9%, 65.0% and 71.0%, respectively. The seroprevalence of HBsAg was 0% in blood donors, 7.0% of visitors for STI, 7.5% of HIV infected individuals and 1.2% in CSWs. That of anti HCV was 0%, 2.5%, 5.0% and 10.3%, respectively. This study underlines the very high seroprevalence of STVs, especially HSV-2, in the group at high risk of STI. The prevalences of HCV in CSWs and HIV infected individuals were two to three times higher than STI patients. It means sexual transmission might be a possible route of transmission of HCV, because HIV infection is closely related with sexual behaviour in Korea. The spread of HSV-2 is dependent on sexual transmission and that the presence of antibody to HSV-2 may be suitable for use as a serological marker of the degree at risk of STI. PMID- 12590794 TI - Can a busy abortion service cope with a screen-and-treat policy for Chlamydia trachomatis infection? AB - In order to determine the effectiveness of a screen-and-treat policy for Chlamydia trachomatis within a busy medical and surgical abortion service, a retrospective audit was conducted of 2058 women undergoing induced abortion over 12 months. The prevalence of C. trachomatis was 6%. Although most positive results were available before a surgical abortion (97%), only 76% were available before a medical abortion (P = 0.007). This resulted in more treatment delays in the medical group (P = 0.04). Although the majority of women (94% surgical and 84% medical) were seen by genitourinary medicine, only one-quarter of partners attended for contact tracing. The current screen-and-treat policy has been shown to be deficient in several areas. Of most concern are the treatment delays with medical abortion, which may be due to the faster 'processing' of women since this method is only available at < or = 9 weeks' gestation. Such challenges to a screen-and-treat policy, are likely to become more common as medical abortion becomes more widely adopted. PMID- 12590795 TI - An audit to evaluate the accessibility, cost, impact on work place absence and convenience of attending genitourinary medicine clinics in London and Plymouth. AB - We conducted a survey to assess the accessibility, cost, impact on work place absence and convenience of attending two genitourinary medicine (GUM) clinics, one in London and one in Plymouth. Nine hundred and fifty-eight questionnaires were collected for analysis (448 from London [L] and 510 from Plymouth [P]). The majority in London used public transport whereas the majority in Plymouth used private transport. Journey costs were less than 5 for the majority (75% L and 59% P). Of those in employment 229/678 (44%) had to take time off from work to attend. Women were significantly less likely (13/308, 4%, 95% confidence interval (CI) 2-6.5%) than men (51/370, 14% 95% [CI] 10.317.3%) to give up annual leave to attend. Both clinics were judged accessible, affordable and convenient by the majority of clinic users. PMID- 12590796 TI - Third regional audit of antiretroviral prescribing in HIV patients. AB - A multicentre re-audit of antiretroviral therapy (ART) prescribing in HIV positive patients from October 2000 to March 2001, in the Yorkshire region, showed an improvement since our previous audit in 1999, with an increased proportion of patients taking three or more drugs. Overall, 53% of all patients on ART had complete HIV-1 virological suppression (<50 copies/mL). It is of concern that only 42% of ART-naive patients who commenced their first regimen during the audit period, achieved complete virological suppression at 24 weeks of follow up. All clinics used the BHIVA guidelines and had access to HIV-1 ultrasensitive viral load and genotypic resistance testing. Future audits will be performed with an emphasis on adherence in relation to treatment response. PMID- 12590797 TI - Low dose interferon-alpha therapy for HIV-associated multicentric Castleman's disease. AB - Successful maintenance therapy with weekly interferon-alpha for HIV-associated multicentric Castleman's disease is described. PMID- 12590798 TI - Sexual dysfunction in HIV-positive individuals. PMID- 12590799 TI - Does discontinuing the use of pill boxes to facilitate electronic monitoring impede adherence? PMID- 12590800 TI - Audit of hepatitis B immunization. PMID- 12590801 TI - Associations of newly diagnosed HIV-1 infections and ethnicity at a provincial UK genitourinary medicine clinic: 1998-2001. PMID- 12590802 TI - Telemedicine and its use in promoting sexual health. PMID- 12590803 TI - Pelvic infection guideline: do we have a consensus? PMID- 12590805 TI - HIV and sexual dysfunction. PMID- 12590806 TI - How is gonorrhoea treated in genitourinary medicine clinics in the UK? PMID- 12590807 TI - Interspike interval correlations, memory, adaptation, and refractoriness in a leaky integrate-and-fire model with threshold fatigue. AB - Neuronal adaptation as well as interdischarge interval correlations have been shown to be functionally important properties of physiological neurons. We explore the dynamics of a modified leaky integrate-and-fire (LIF) neuron, referred to as the LIF with threshold fatigue, and show that it reproduces these properties. In this model, the postdischarge threshold reset depends on the preceding sequence of discharge times. We show that in response to various classes of stimuli, namely, constant currents, step currents, white gaussian noise, and sinusoidal currents, the model exhibits new behavior compared with the standard LIF neuron. More precisely, (1) step currents lead to adaptation, that is, a progressive decrease of the discharge rate following the stimulus onset, while in the standard LIF, no such patterns are possible; (2) a saturation in the firing rate occurs in certain regimes, a behavior not seen in the LIF neuron; (3) interspike intervals of the noise-driven modified LIF under constant current are correlated in a way reminiscent of experimental observations, while those of the standard LIF are independent of one another; (4) the magnitude of the correlation coefficients decreases as a function of noise intensity; and (5) the dynamics of the sinusoidally forced modified LIF are described by iterates of an annulus map, an extension to the circle map dynamics displayed by the LIF model. Under certain conditions, this map can give rise to sensitivity to initial conditions and thus chaotic behavior. PMID- 12590808 TI - Reliability of spike timing is a general property of spiking model neurons. AB - The responses of neurons to time-varying injected currents are reproducible on a trial-by-trial basis in vitro, but when a constant current is injected, small variances in interspike intervals across trials add up, eventually leading to a high variance in spike timing. It is unclear whether this difference is due to the nature of the input currents or the intrinsic properties of the neurons. Neuron responses can fail to be reproducible in two ways: dynamical noise can accumulate over time and lead to a desynchronization over trials, or several stable responses can exist, depending on the initial condition. Here we show, through simulations and theoretical considerations, that for a general class of spiking neuron models, which includes, in particular, the leaky integrate-and fire model as well as nonlinear spiking models, aperiodic currents, contrary to periodic currents, induce reproducible responses, which are stable under noise, change in initial conditions and deterministic perturbations of the input. We provide a theoretical explanation for aperiodic currents that cross the threshold. PMID- 12590809 TI - The dynamic neural filter: a binary model of spatiotemporal coding. AB - We describe and discuss the properties of a binary neural network that can serve as a dynamic neural filter (DNF), which maps regions of input space into spatiotemporal sequences of neuronal activity. Both deterministic and stochastic dynamics are studied, allowing the investigation of the stability of spatiotemporal sequences under noisy conditions. We define a measure of the coding capacity of a DNF and develop an algorithm for constructing a DNF that can serve as a source of given codes. On the basis of this algorithm, we suggest using a minimal DNF capable of generating observed sequences as a measure of complexity of spatiotemporal data. This measure is applied to experimental observations in the locust olfactory system, whose reverberating local field potential provides a natural temporal scale allowing the use of a binary DNF. For random synaptic matrices, a DNF can generate very large cycles, thus becoming an efficient tool for producing spatiotemporal codes. The latter can be stabilized by applying to the parameters of the DNF a learning algorithm with suitable margins. PMID- 12590810 TI - Modeling short-term synaptic depression in silicon. AB - We describe a model of short-term synaptic depression that is derived from a circuit implementation. The dynamics of this circuit model is similar to the dynamics of some theoretical models of short-term depression except that the recovery dynamics of the variable describing the depression is nonlinear and it also depends on the presynaptic frequency. The equations describing the steady state and transient responses of this synaptic model are compared to the experimental results obtained from a fabricated silicon network consisting of leaky integrate-and-fire neurons and different types of short-term dynamic synapses. We also show experimental data demonstrating the possible computational roles of depression. One possible role of a depressing synapse is that the input can quickly bring the neuron up to threshold when the membrane potential is close to the resting potential. PMID- 12590811 TI - Dictionary learning algorithms for sparse representation. AB - Algorithms for data-driven learning of domain-specific overcomplete dictionaries are developed to obtain maximum likelihood and maximum a posteriori dictionary estimates based on the use of Bayesian models with concave/Schur-concave (CSC) negative log priors. Such priors are appropriate for obtaining sparse representations of environmental signals within an appropriately chosen (environmentally matched) dictionary. The elements of the dictionary can be interpreted as concepts, features, or words capable of succinct expression of events encountered in the environment (the source of the measured signals). This is a generalization of vector quantization in that one is interested in a description involving a few dictionary entries (the proverbial "25 words or less"), but not necessarily as succinct as one entry. To learn an environmentally adapted dictionary capable of concise expression of signals generated by the environment, we develop algorithms that iterate between a representative set of sparse representations found by variants of FOCUSS and an update of the dictionary using these sparse representations. Experiments were performed using synthetic data and natural images. For complete dictionaries, we demonstrate that our algorithms have improved performance over other independent component analysis (ICA) methods, measured in terms of signal-to-noise ratios of separated sources. In the overcomplete case, we show that the true underlying dictionary and sparse sources can be accurately recovered. In tests with natural images, learned overcomplete dictionaries are shown to have higher coding efficiency than complete dictionaries; that is, images encoded with an overcomplete dictionary have both higher compression (fewer bits per pixel) and higher accuracy (lower mean square error). PMID- 12590813 TI - Linear geometric ICA: fundamentals and algorithms. AB - Geometric algorithms for linear independent component analysis (ICA) have recently received some attention due to their pictorial description and their relative ease of implementation. The geometric approach to ICA was proposed first by Puntonet and Prieto (1995). We will reconsider geometric ICA in a theoretic framework showing that fixed points of geometric ICA fulfill a geometric convergence condition (GCC), which the mixed images of the unit vectors satisfy too. This leads to a conjecture claiming that in the nongaussian unimodal symmetric case, there is only one stable fixed point, implying the uniqueness of geometric ICA after convergence. Guided by the principles of ordinary geometric ICA, we then present a new approach to linear geometric ICA based on histograms observing a considerable improvement in separation quality of different distributions and a sizable reduction in computational cost, by a factor of 100, compared to the ordinary geometric approach. Furthermore, we explore the accuracy of the algorithm depending on the number of samples and the choice of the mixing matrix, and compare geometric algorithms with classical ICA algorithms, namely, Extended Infomax and FastICA. Finally, we discuss the problem of high-dimensional data sets within the realm of geometrical ICA algorithms. PMID- 12590814 TI - Equivalence of backpropagation and contrastive Hebbian learning in a layered network. AB - Backpropagation and contrastive Hebbian learning are two methods of training networks with hidden neurons. Backpropagation computes an error signal for the output neurons and spreads it over the hidden neurons. Contrastive Hebbian learning involves clamping the output neurons at desired values and letting the effect spread through feedback connections over the entire network. To investigate the relationship between these two forms of learning, we consider a special case in which they are identical: a multilayer perceptron with linear output units, to which weak feedback connections have been added. In this case, the change in network state caused by clamping the output neurons turns out to be the same as the error signal spread by backpropagation, except for a scalar prefactor. This suggests that the functionality of backpropagation can be realized alternatively by a Hebbian-type learning algorithm, which is suitable for implementation in biological networks. PMID- 12590812 TI - Spatiochromatic receptive field properties derived from information-theoretic analyses of cone mosaic responses to natural scenes. AB - Neurons in the early stages of processing in the primate visual system efficiently encode natural scenes. In previous studies of the chromatic properties of natural images, the inputs were sampled on a regular array, with complete color information at every location. However, in the retina cone photoreceptors with different spectral sensitivities are arranged in a mosaic. We used an unsupervised neural network model to analyze the statistical structure of retinal cone mosaic responses to calibrated color natural images. The second order statistical dependencies derived from the covariance matrix of the sensory signals were removed in the first stage of processing. These decorrelating filters were similar to type I receptive fields in parvo- or konio-cellular LGN in both spatial and chromatic characteristics. In the subsequent stage, the decorrelated signals were linearly transformed to make the output as statistically independent as possible, using independent component analysis. The independent component filters showed luminance selectivity with simple-cell-like receptive fields, or had strong color selectivity with large, often double opponent, receptive fields, both of which were found in the primary visual cortex (V1). These results show that the "form" and "color" channels of the early visual system can be derived from the statistics of sensory signals. PMID- 12590815 TI - Indexed families of functionals and gaussian radial basis functions. AB - We report on results concerning the capabilities of gaussian radial basis function networks in the setting of inner product spaces that need not be finite dimensional. Specifically, we show that important indexed families of functionals can be uniformly approximated, with the approximation uniform also with respect to the index. Applications are described concerning the classification of signals and the synthesis of reconfigurable classifiers. PMID- 12590816 TI - Efficient greedy learning of gaussian mixture models. AB - This article concerns the greedy learning of gaussian mixtures. In the greedy approach, mixture components are inserted into the mixture one after the other. We propose a heuristic for searching for the optimal component to insert. In a randomized manner, a set of candidate new components is generated. For each of these candidates, we find the locally optimal new component and insert it into the existing mixture. The resulting algorithm resolves the sensitivity to initialization of state-of-the-art methods, like expectation maximization, and has running time linear in the number of data points and quadratic in the (final) number of mixture components. Due to its greedy nature, the algorithm can be particularly useful when the optimal number of mixture components is unknown. Experimental results comparing the proposed algorithm to other methods on density estimation and texture segmentation are provided. PMID- 12590817 TI - SMO algorithm for least-squares SVM formulations. AB - This article extends the well-known SMO algorithm of support vector machines (SVMs) to least-squares SVM formulations that include LS-SVM classification, kernel ridge regression, and a particular form of regularized kernel Fisher discriminant. The algorithm is shown to be asymptotically convergent. It is also extremely easy to implement. Computational experiments show that the algorithm is fast and scales efficiently (quadratically) as a function of the number of examples. PMID- 12590818 TI - Asynchronous states and the emergence of synchrony in large networks of interacting excitatory and inhibitory neurons. AB - We investigate theoretically the conditions for the emergence of synchronous activity in large networks, consisting of two populations of extensively connected neurons, one excitatory and one inhibitory. The neurons are modeled with quadratic integrate-and-fire dynamics, which provide a very good approximation for the subthreshold behavior of a large class of neurons. In addition to their synaptic recurrent inputs, the neurons receive a tonic external input that varies from neuron to neuron. Because of its relative simplicity, this model can be studied analytically. We investigate the stability of the asynchronous state (AS) of the network with given average firing rates of the two populations. First, we show that the AS can remain stable even if the synaptic couplings are strong. Then we investigate the conditions under which this state can be destabilized. We show that this can happen in four generic ways. The first is a saddle-node bifurcation, which leads to another state with different average firing rates. This bifurcation, which occurs for strong enough recurrent excitation, does not correspond to the emergence of synchrony. In contrast, in the three other instability mechanisms, Hopf bifurcations, which correspond to the emergence of oscillatory synchronous activity, occur. We show that these mechanisms can be differentiated by the firing patterns they generate and their dependence on the mutual interactions of the inhibitory neurons and cross talk between the two populations. We also show that besides these codimension 1 bifurcations, the system can display several codimension 2 bifurcations: Takens Bogdanov, Gavrielov-Guckenheimer, and double Hopf bifurcations. PMID- 12590819 TI - A constrained EM algorithm for principal component analysis. AB - We propose a constrained EM algorithm for principal component analysis (PCA) using a coupled probability model derived from single-standard factor analysis models with isotropic noise structure. The single probabilistic PCA, especially for the case where there is no noise, can find only a vector set that is a linear superposition of principal components and requires postprocessing, such as diagonalization of symmetric matrices. By contrast, the proposed algorithm finds the actual principal components, which are sorted in descending order of eigenvalue size and require no additional calculation or postprocessing. The method is easily applied to kernel PCA. It is also shown that the new EM algorithm is derived from a generalized least-squares formulation. PMID- 12590820 TI - Higher-order statistics of input ensembles and the response of simple model neurons. AB - Pairwise correlations among spike trains recorded in vivo have been frequently reported. It has been argued that correlated activity could play an important role in the brain, because it efficiently modulates the response of a postsynaptic neuron. We show here that a neuron's output firing rate critically depends on the higher-order statistics of the input ensemble. We constructed two statistical models of populations of spiking neurons that fired with the same rates and had identical pairwise correlations, but differed with regard to the higher-order interactions within the population. The first ensemble was characterized by clusters of spikes synchronized over the whole population. In the second ensemble, the size of spike clusters was, on average, proportional to the pairwise correlation. For both input models, we assessed the role of the size of the population, the firing rate, and the pairwise correlation on the output rate of two simple model neurons: a continuous firing-rate model and a conductance-based leaky integrate-and-fire neuron. An approximation to the mean output rate of the firing-rate neuron could be derived analytically with the help of shot noise theory. Interestingly, the essential features of the mean response of the two neuron models were similar. For both neuron models, the three input parameters played radically different roles with respect to the postsynaptic firing rate, depending on the interaction structure of the input. For instance, in the case of an ensemble with small and distributed spike clusters, the output firing rate was efficiently controlled by the size of the input population. In addition to the interaction structure, the ratio of inhibition to excitation was found to strongly modulate the effect of correlation on the postsynaptic firing rate. PMID- 12590821 TI - Duality of rate coding and temporal coding in multilayered feedforward networks. AB - A functional role for precise spike timing has been proposed as an alternative hypothesis to rate coding. We show in this article that both the synchronous firing code and the population rate code can be used dually in a common framework of a single neural network model. Furthermore, these two coding mechanisms are bridged continuously by several modulatable model parameters, including shared connectivity, feedback strength, membrane leak rate, and neuron heterogeneity. The rates of change of these parameters are closely related to the response time and the timescale of learning. PMID- 12590822 TI - Synchronous firing and higher-order interactions in neuron pool. AB - The stochastic mechanism of synchronous firing in a population of neurons is studied from the point of view of information geometry. Higher-order interactions of neurons, which cannot be reduced to pairwise correlations, are proved to exist in synchronous firing. In a neuron pool where each neuron fires stochastically, the probability distribution q(r) of the activity r, which is the fraction of firing neurons in the pool, is studied. When q(r) has a widespread distribution, in particular, when q(r) has two peaks, the neurons fire synchronously at one time and are quiescent at other times. The mechanism of generating such a probability distribution is interesting because the activity r is concentrated on its mean value when each neuron fires independently, because of the law of large numbers. Even when pairwise interactions, or third-order interactions, exist, the concentration is not resolved. This shows that higher-order interactions are necessary to generate widespread activity distributions. We analyze a simple model in which neurons receive common overlapping inputs and prove that such a model can have a widespread distribution of activity, generating higher-order stochastic interactions. PMID- 12590823 TI - Determination of firing times for the stochastic Fitzhugh-Nagumo neuronal model. AB - We present for the first time an analytical approach for determining the time of firing of multicomponent nonlinear stochastic neuronal models. We apply the theory of first exit times for Markov processes to the Fitzhugh-Nagumo system with a constant mean gaussian white noise input, representing stochastic excitation and inhibition. Partial differential equations are obtained for the moments of the time to first spike. The observation that the recovery variable barely changes in the prespike trajectory leads to an accurate one-dimensional approximation. For the moments of the time to reach threshold, this leads to ordinary differential equations that may be easily solved. Several analytical approaches are explored that involve perturbation expansions for large and small values of the noise parameter. For ranges of the parameters appropriate for these asymptotic methods, the perturbation solutions are used to establish the validity of the one-dimensional approximation for both small and large values of the noise parameter. Additional verification is obtained with the excellent agreement between the mean and variance of the firing time found by numerical solution of the differential equations for the one-dimensional approximation and those obtained by simulation of the solutions of the model stochastic differential equations. Such agreement extends to intermediate values of the noise parameter. For the mean time to threshold, we find maxima at small noise values that constitute a form of stochastic resonance. We also investigate the dependence of the mean firing time on the initial values of the voltage and recovery variables when the input current has zero mean. PMID- 12590825 TI - A quantified sensitivity measure for multilayer perceptron to input perturbation. AB - The sensitivity of a neural network's output to its input perturbation is an important issue with both theoretical and practical values. In this article, we propose an approach to quantify the sensitivity of the most popular and general feedforward network: multilayer perceptron (MLP). The sensitivity measure is defined as the mathematical expectation of output deviation due to expected input deviation with respect to overall input patterns in a continuous interval. Based on the structural characteristics of the MLP, a bottom-up approach is adopted. A single neuron is considered first, and algorithms with approximately derived analytical expressions that are functions of expected input deviation are given for the computation of its sensitivity. Then another algorithm is given to compute the sensitivity of the entire MLP network. Computer simulations are used to verify the derived theoretical formulas. The agreement between theoretical and experimental results is quite good. The sensitivity measure can be used to evaluate the MLP's performance. PMID- 12590824 TI - Developmental constraints aid the acquisition of binocular disparity sensitivities. AB - This article considers the hypothesis that systems learning aspects of visual perception may benefit from the use of suitably designed developmental progressions during training. We report the results of simulations in which four models were trained to detect binocular disparities in pairs of visual images. Three of the models were developmental models in the sense that the nature of their visual input changed during the course of training. These models received a relatively impoverished visual input early in training, and the quality of this input improved as training progressed. One model used a coarse-scale-to multiscale developmental progression, another used a fine-scale-to-multiscale progression, and the third used a random progression. The final model was nondevelopmental in the sense that the nature of its input remained the same throughout the training period. The simulation results show that the two developmental models whose progressions were organized by spatial frequency content consistently outperformed the nondevelopmental and random developmental models. We speculate that the superior performance of these two models is due to two important features of their developmental progressions: (1) these models were exposed to visual inputs at a single scale early in training, and (2) the spatial scale of their inputs progressed in an orderly fashion from one scale to a neighboring scale during training. Simulation results consistent with these speculations are presented. We conclude that suitably designed developmental sequences can be useful to systems learning to detect binocular disparities. The idea that visual development can aid visual learning is a viable hypothesis in need of study. PMID- 12590826 TI - Variational mixture of Bayesian independent component analyzers. AB - There has been growing interest in subspace data modeling over the past few years. Methods such as principal component analysis, factor analysis, and independent component analysis have gained in popularity and have found many applications in image modeling, signal processing, and data compression, to name just a few. As applications and computing power grow, more and more sophisticated analyses and meaningful representations are sought. Mixture modeling methods have been proposed for principal and factor analyzers that exploit local gaussian features in the subspace manifolds. Meaningful representations may be lost, however, if these local features are nongaussian or discontinuous. In this article, we propose extending the gaussian analyzers mixture model to an independent component analyzers mixture model. We employ recent developments in variational Bayesian inference and structure determination to construct a novel approach for modeling nongaussian, discontinuous manifolds. We automatically determine the local dimensionality of each manifold and use variational inference to calculate the optimum number of ICA components needed in our mixture model. We demonstrate our framework on complex synthetic data and illustrate its application to real data by decomposing functional magnetic resonance images into meaningful-and medically useful-features. PMID- 12590827 TI - Effects of reversible cold block of face primary somatosensory cortex on orofacial movements and related face primary motor cortex neuronal activity. AB - Our previous studies have revealed that face primary somatosensory cortex (SI) as well as face primary motor cortex (MI) play important roles in the control of orofacial movements in awake monkeys, and that both face MI and face SI neurons may have an orofacial mechanoreceptive field and show activity related to orofacial movements. Since it is possible that the movement-related activity of face MI neurons could reflect movement-generated orofacial afferent inputs projecting to face MI via face SI, the present study used reversible cold block induced inactivation of the monkey's face SI to determine if face MI neuronal activity related to a trained tongue-protrusion task, chewing or swallowing was dependent on the functional integrity of the ipsilateral face SI and if inactivation of face SI affects orofacial movements. The effects of face SI cold block were tested on chewing, swallowing and/or task-related activity of 73 face MI neurons. Both task and chewing and/or swallowing-related activity of most face MI neurons was independent of the functional integrity of the ipsilateral face SI since SI cold block affected the movement-related activity in approximately 25% of the neurons. Similarly, unilateral cold block of SI had very limited effects on the performance of the task and chewing, and no effect on the performance of swallowing. These findings suggest that movement-induced reafferentation via face SI may not be a significant factor in accounting for the activity of the majority of ipsilateral face MI neurons related to trained movements, chewing and swallowing. PMID- 12590828 TI - Effects of stimulus duration on neuronal response properties in the somatosensory cortex of the star-nosed mole. AB - Star-nosed moles have a series of mechanosensory appendages surrounding each nostril. Each appendage is covered with sensory organs (Eimer's organs) containing both rapidly adapting and slowly adapting mechanoreceptors and each appendage is represented in primary somatosensory cortex (S1) by a single cortical module. When the skin surface of an appendage is depressed, neurons in the corresponding module in S1 respond in either a transient or sustained fashion. The aim of this study was to characterize and compare the responses of these two classes of neurons to both short (5 or 20 ms) and long (500 ms) mechanosensory stimulation. Activity from neurons in the representation of appendage 11, the somatosensory fovea, was recorded while delivering mechanosensory stimuli to the corresponding skin surface. Transient and sustained neurons had different levels of spontaneous activity and different responses to both short and long mechanosensory stimulation. Neurons with sustained responses had a significantly higher spontaneous firing rate than neurons with transient responses. Transient neurons responded to a 5 ms stimulus with excitation followed by suppression of discharge whereas sustained neurons did not exhibit post-excitatory suppression. Rather, responses of sustained neurons to 5 ms stimuli lasted several hundred milliseconds. Consequently sustained responses contained significantly more spikes than transient responses. These experiments suggest contact to the appendages causes two distinct firing patterns in cortex regardless of the duration of the stimulus. The sustained and transient responses could reflect either the activity of fundamentally different classes of neurons or activity in distinct subcortical and cortical networks. PMID- 12590829 TI - The role of sensory input in motor neuron sprouting control. AB - Primary sensory neurons project to motor neurons directly or through interneurons and affect their activity. In our previous paper we showed that intramuscular sprouting can be affected by changing the sensory synaptic input to motor neurons. In this work, motor axon sprouting within a peripheral nerve (extramuscular sprouting) was induced by nerve injury at such a distance from muscle so as not to allow nerve-muscle trophic interactions. Two different procedures were carried out: (1) sciatic nerve crush and (2) sciatic nerve crush with homosegmental ipsilateral L3-L5 dorsal rhizotomy. The number of regenerating motor axons innervating extensor digitorum longus muscle was determined by in vivo muscle tension recordings and an index of their individual conduction rate was obtained by in vitro intracellular recordings of excitatory postsynaptic end plate potentials in muscle fibers. The main findings were: (1) there are more regenerated axons distally from the lesion than parent axons proximally to the lesion (sprouting at the lesion); (2) sprouting at the lesion was negatively affected by homosegmental ipsilateral dorsal rhizotomy; (3) the number of motor axons innervating extensor digitorum longus muscle extrafusal fibers counted proximally to the lesion increased following nerve injury and regeneration but this did not occur when sensory input was lost. A transient innervation of extrafusal fibers by gamma motor neurons may explain the increase of motor axons counted proximally to the lesion. PMID- 12590830 TI - The influence of increased muscle spindle sensitivity on Achilles tendon jerk and H-reflex in relaxed human subjects. AB - Whether the fusimotor system contributes to reflex gain changes during reinforcement maneuvers is re-examined in the light of new data. Recently, from direct recordings of spindle afferent activity originating from ankle flexor muscles, we showed that mental computation increased the muscle spindle mechanical sensitivity in completely relaxed human subjects without concomitant alpha-motoneuron activation, providing evidence for selective fusimotor drive activation. In the present study, the effects of mental computation were investigated on monosynaptic reflexes elicited in non-contracting soleus muscle either by direct nerve stimulation (Hoffmann reflex, H) or by tendon tap (Tendinous reflex, T). The aim was to relate the time course of the changes in reflex size to the increase in spindle sensitivity during mental task in order to explore whether fusimotor activation can influence the size of the monosynaptic reflex. The results show changes in reflex amplitude that parallel the increase in muscle spindle sensitivity. When T-reflex is consistently facilitated during mental effort, the H-reflex is either depressed or facilitated, depending on the subjects. These findings suggest that the increased activity in muscle spindle primary endings may account for mental computation-induced changes in both tendon jerk and H-reflex. The facilitation of T-reflex is attributed to the enhanced spindle mechanical sensitivity and the inhibition of H-reflex is attributed to post-activation depression following the increased Ia ongoing discharge. This study supports the view that the fusimotor sensitization of muscle spindles is responsible for changes in both the mechanically and electrically elicited reflexes. It is concluded that the fusimotor drive contributed to adjustment of the size of tendon jerk and H-reflex during mental effort. The possibility that a mental computation task may also operate by reducing the level of presynaptic inhibition is discussed on the basis of H-reflex facilitation. PMID- 12590831 TI - A glimpse of the brain transforming a sensory signal into a motor response. AB - Averages were made of neuronal spike activity recorded successively from eight relay regions along the auditorimotor pathway of naive cats and cats conditioned to blink in response to a 70 dB click conditioned stimulus (CS). It was hypothesized that the patterns of activity could be distinguished as sensory or motor by differences in their relationship to the pattern of the acoustic CS vs that of the conditioned response (CR). If so, it was also hypothesized that the acoustic stimulus would be better expressed at early auditorimotor relays and the motor response at later relays along the pathway. To test these hypotheses, Pearson correlation coefficients were calculated between the mean patterns of unit activity at each of the auditorimotor relays and (1) the rectified sound pattern of the CS and (2) the averaged, rectified electromyographic (EMG) activity of the muscles (orbicularis oculis) that produced the CR. In both naive and conditioned cats, there were significant positive correlations between the patterns of spike activity and the sound at early relays along the auditorimotor pathway such as the cochlear nucleus and inferior colliculus. In the conditioned animals, the spike activity of later nuclei in the auditorimotor pathway, such as the rostral thalamus and the motor cortex, had the highest positive correlations with the motor response. These correlations were low in the naive animals. Thus, the mean patterns of spike activity along the auditorimotor pathway appeared to distinguish the sound from the motor response and provided a glimpse of the process supporting transformation of the CS into the incipient CR. PMID- 12590832 TI - The genitals and gluteal skin are represented lateral to the foot in anterior parietal somatosensory cortex of macaques. AB - Detailed electrophysiological maps of the representations of trunk and adjacent body parts in area 3b and area 1 of somatosensory cortex were obtained in three macaque monkeys (Macaca mulatta and Macaca radiata) of either sex. A total of 211 microelectrode penetrations 250-300 microm apart resulted in 1,190 recording sites. During penetrations deep into the posterior bank of the central sulcus, recordings were made every 300 microm to depths of 6-7 mm until sites unresponsive to somatic stimuli were reached. Cortex was later cut parasagittally and sections were stained for cytochrome oxidase (CO) or Nissl substance. Contrary to expectations from earlier reports, the genitalia were represented lateral to the representations of the foot in cortex along the area 3b/1 border. The gluteal skin including the gluteal pads and the base of the tail were also represented in this section of cortex. Only a small region of cortex was devoted to the genitalia, and neurons in this cortex had receptive fields that were large and typically included skin of the inner thigh and belly. The lower, middle and upper trunk were represented more laterally, followed by the neck, upper head and arm. The receptive fields on the trunk were roughly the same size as those for the middle and lower trunk and slightly smaller on the upper trunk. PMID- 12590833 TI - Diabetic neuropathy and surface sway-referencing disrupt somatosensory information for postural stability in stance. AB - In order to determine the type of somatosensory information for postural control that is most affected by neuropathy, we compared the relative effects of three methods of sway-referencing the surface in a group of subjects with profound loss of somatosensory function associated with sensory polyneuropathy from diabetes with age-matched control subjects. Sway-referencing disrupted somatosensory feedback for postural control by servo-controlling the dorsi- and plantar-flexion rotation of the support surface in proportion to anterior-posterior excursion of (1) ankle angle, (2) center of body mass (CoM) angle or (3) filtered center of pressure (CoP). Postural sway in subjects with somatosensory loss was significantly larger than normal on a firm surface but not on the sway-referenced surfaces, suggesting that sway-referencing disrupts somatosensory information for postural control already disrupted by neuropathy. Control subjects standing on any sway-referenced surface swayed significantly more than neuropathy subjects who stood on a firm surface, suggesting that sway-referencing disrupts more somatosensory information than disrupted by severe neuropathy. CoP sway referencing was less sensitive than ankle or CoM sway-referencing for distinguishing postural sway in subjects with somatosensory loss from age-matched control subjects. Given that filtered CoP sway-referencing disrupts the ability to utilize somatosensory information related to surface reactive force to a greater extent than the other two methods of sway-referencing, then these results support the hypothesis that subjects with diabetic peripheral neuropathy have lost more CoP information, than ankle or CoM angle information, for controlling postural sway in stance. PMID- 12590834 TI - Muscular torque generation during imposed joint rotation: torque-angle relationships when subjects' only goal is to make a constant effort. AB - It is a reasonable expectation that voluntarily activated spinal motoneurons will be further excited by increases in spindle afferent activity produced by muscle stretch. Human motor behavior attributed to tonic stretch reflexes and to reflexes recruited by relatively slow joint rotation has been reported from several laboratories. We reinvestigated this issue by rotating the elbow joint over the central portion of its range while subjects focused on keeping their elbow flexion effort constant at one of three different levels and made no attempt to control the position, speed or direction of movement of their forearm. There is evidence that subjects' voluntary motor status is constant under these conditions so that any change in torque would be of involuntary origin. On average, torques rose somewhat and then fell as the elbow was flexed through a range of 80 degrees at 10, 20 and 60 degrees/s and a similar pattern occurred during elbow extension; i.e., both concentric and eccentric torque-angle profiles had roughly similar shapes and neither produced consistent stabilizing cross range stiffness. The negative stiffness (rising torque) during the early part of a concentric movement and the negative stiffness (falling torque) during the later part of an eccentric movement would not have occurred if a stabilizing stretch reflex had been present. Positive stiffness rarely gave rise to torque changes greater than 20% in either individual or cross-subject averaged data. When angular regions of negative stiffness are combined with regions of low positive stiffness (torque change 10% or less), much of the range of motion was not well stabilized, especially during eccentric movements. The sum of the EMGs from biceps brachii, brachioradialis and brachialis showed a pattern opposite to that expected for a stretch reflex; there was an upward trend in the EMG as the elbow was flexed and a downward trend as the elbow was extended. There was little change in the shape of this EMG-angle relationship with either direction or velocity. The individual EMG-angle relationships were distinctive for each of these three elbow flexor muscles in four of the six subjects; in the remaining two, biceps was distinctive, but brachioradialis and brachialis appeared to be coupled. Although the EMGs of individual muscles were modulated over the angular range, no consistent stretch reflexes could be seen in the individual records. Thus, we could find no clear evidence for stretch reflex stabilization of human subjects maintaining a constant effort. Rather, muscle torque appears to be reflexly modulated across a much used portion of the elbow's angular range so that any appreciable stabilizing stiffness that is sustained for more than fractions of a second is associated with a change in effort. PMID- 12590835 TI - Topography of rodent whisking--I. Two-dimensional monitoring of whisker movements. AB - During 'active touch' the rodent whiskers scan the environment in a series of repetitive movements ('whisks') generating afferent signals which transform the spatial properties of objects into spatio-temporal patterns of neural activity. Based upon analyses carried out in a single movement plane, it has been generally assumed that these trajectories are essentially uni-dimensional, although more complex movements have been described in some rodents. The present study was designed to examine this assumption and to more precisely characterize whisking topography by monitoring whisking trajectories along both the antero-posterior and dorso-ventral axes. Using optoelectronic monitoring techniques with high spatio-temporal resolution, movement data were obtained from a population of vibrissae sampled at different locations on the mystacial pad in head-fixed rats isolated from the perturbing effects of contact. For a substantial proportion of the population of whisking movements sampled, the trajectories generated by a single whisker is most accurately described as occupying an expended two dimensional space in which the A-P component predominates. However, the whisker system exhibits a considerable range of trajectory types, suggesting a high degree of movement flexibility. For each vibrissa position, it was possible to delineate a 'trajectory' domain-that portion of the animal's whisking space which is scanned by the movements of that vibrissa during whisking. Since the 'domains' of adjacent whiskers in the same row tend to overlap, synchronized movements of a subset of whiskers could generate a set of overlapping somatosensory fields analogous to overlapping retinal receptive fields. The organization of such trajectory domains within the rats' whisking space could provide the spatial component of the spatio-temporal integration process required to extract information about environmental features from the inputs generated by its recursive whisking movements. PMID- 12590837 TI - Reversible blockade of rodent whisking: Botulinum toxin as a tool for developmental studies. AB - Studies of sensorimotor systems such as the whisking system of rodents have suggested that associations between body movements and their sensory consequences during development may make an important contribution to the functional organization of the system. In the present study we have explored the possible utility of Botulinum toxin for developmental studies of whisking. Botox selectively blocks whisking-generated afference leaving other sources of whisker afference intact. We describe appropriate modes of injection, define dosage levels, and assess the effects of prolonged whisking paralysis during development upon the basic motor competency of the adult rat. Our findings indicate that: (a) Botulinum toxin may be used to block whisking behavior in adult and developing rats, (b) that the duration of the whisking paralysis produced by Botox treatment blockade is dose dependent in both developing and adult animals, (c) that the blockade is functionally reversible and (d) that Botox treatment during development does not impair either the kinematics or the rhythmic patterning of adult whisking behavior. Botox may be a useful tool for studying the role of experiential factors in the development of "active touch" in rodents. PMID- 12590836 TI - Sensory representation abnormalities that parallel focal hand dystonia in a primate model. AB - In our hypothesis of focal dystonia, attended repetitive behaviors generate aberrant sensory representations. Those aberrant representations interfere with motor control. Abnormal motor control strengthens sensory abnormalities. The positive feedback loop reinforces the dystonic condition. Previous studies of primates with focal hand dystonia have demonstrated multi-digit or hairy-glabrous responses at single sites in area 3b, receptive fields that average ten times larger than normal, and high receptive field overlap as a function of horizontal distance. In this study, we strengthen and elaborate these findings. One animal was implanted with an array of microelectrodes that spanned the border between the face and digits. After the animal developed hand dystonia, responses in the initial hand representation increasingly responded to low threshold stimulation of the face in a columnar substitution. The hand-face border that is normally sharp became patchy and smeared over 1 mm of cortex within 6 weeks. Two more trained animals developed a focal hand dystonia variable in severity across the hand. Receptive field size, presence of multi-digit or hairy-glabrous receptive fields, and columnar overlap covaried with the animal's ability to use specific digits. A fourth animal performed the same behaviors without developing dystonia. Many of its physiological measures were similar to the dystonic animals, but receptive field overlap functions were minimally abnormal, and no sites shared response properties that are normally segregated such as hairy-glabrous combined fields, or multi-digit fields. Thalamic mapping demonstrated proportionate levels of abnormality in thalamic representations as were found in cortical representations. PMID- 12590838 TI - Dreaming and episodic memory: a functional dissociation? AB - The activity that takes place in memory systems during sleep is likely to be related to the role of sleep in memory consolidation and learning, as well as to the generation of dream hallucinations. This study addressed the often-stated hypothesis that replay of whole episodic memories contributes to the multimodal hallucinations of sleep. Over a period of 14 days, 29 subjects kept a log of daytime activities, events, and concerns, wrote down any recalled dreams, and scored the dreams for incorporation of any waking experiences. While 65% of a total of 299 sleep mentation reports were judged to reflect aspects of recent waking life experiences, the episodic replay of waking events was found in no more than 1-2% of the dream reports. This finding has implications for understanding the unique memory processing that takes place during the night and is consistent with evidence that sleep has no role in episodic memory consolidation. PMID- 12590839 TI - Neural substrates of perceptual enhancement by cross-modal spatial attention. AB - Orienting attention involuntarily to the location of a sudden sound improves perception of subsequent visual stimuli that appear nearby. The neural substrates of this cross-modal attention effect were investigated by recording event-related potentials to the visual stimuli using a dense electrode array and localizing their brain sources through inverse dipole modeling. A spatially nonpredictive auditory precue modulated visual-evoked neural activity first in the superior temporal cortex at 120-140 msec and then in the ventral occipital cortex of the fusiform gyrus 15-25 msec later. This spatio-temporal sequence of brain activity suggests that enhanced visual perception produced by the cross-modal orienting of spatial attention results from neural feedback from the multimodal superior temporal cortex to the visual cortex of the ventral processing stream. PMID- 12590840 TI - Visual localization ability influences cross-modal bias. AB - The ability of a visual signal to influence the localization of an auditory target (i.e., "cross-modal bias") was examined as a function of the spatial disparity between the two stimuli and their absolute locations in space. Three experimental issues were examined: (a) the effect of a spatially disparate visual stimulus on auditory localization judgments; (b) how the ability to localize visual, auditory, and spatially aligned multisensory (visual-auditory) targets is related to cross-modal bias, and (c) the relationship between the magnitude of cross-modal bias and the perception that the two stimuli are spatially "unified" (i.e., originate from the same location). Whereas variability in localization of auditory targets was large and fairly uniform for all tested locations, variability in localizing visual or spatially aligned multisensory targets was much smaller, and increased with increasing distance from the midline. This trend proved to be strongly correlated with biasing effectiveness, for although visual auditory bias was unexpectedly large in all conditions tested, it decreased progressively (as localization variability increased) with increasing distance from the midline. Thus, central visual stimuli had a substantially greater biasing effect on auditory target localization than did more peripheral visual stimuli. It was also apparent that cross-modal bias decreased as the degree of visual-auditory disparity increased. Consequently, the greatest visual-auditory biases were obtained with small disparities at central locations. In all cases, the magnitude of these biases covaried with judgments of spatial unity. The results suggest that functional properties of the visual system play the predominant role in determining these visual-auditory interactions and that cross modal biases can be substantially greater than previously noted. PMID- 12590841 TI - Actions speak louder than functions: the importance of manipulability and action in tool representation. AB - PET was used to investigate the neural correlates of action knowledge in object representations, particularly the left lateralized network of activations previously implicated in the processing of tools and their associated actions: ventral premotor cortex (VPMCx), posterior middle temporal gyrus (PMTG), and intraparietal sulcus (IPS). Judgments were made about the actions and functions associated with manipulable man-made objects (e.g., hammer); this enabled us to measure activations in response to both explicit and implicit retrieval of knowledge about actions associated with manipulable tools. Function judgments were also made about nonmanipulable artifacts (e.g., traffic light) providing a direct comparison for manipulable objects. Although neither the left VPMCx nor the left PMTG were selective for tool stimuli (nonmanipulable objects also activated these areas relative to a visual control condition), both regions responded more strongly to manipulable objects, suggesting a role for these cortical areas in the processing of knowledge associated with tools. Furthermore, these activations were insensitive to retrieval task, suggesting that visually presented tools automatically recruit both left VPMCx and left PMTG in response to action features that are inherent in tool representations. In contrast, the IPS showed clear selectivity for explicit retrieval of action information about manipulable objects. No regions of cortex were more activated by function relative to action judgments about artifacts. These results are consistent with the brain's preferential responsiveness to how we interact with objects, rather than what they are used for. PMID- 12590842 TI - Parietal representation of symbolic and nonsymbolic magnitude. AB - The close behavioral parallels between the processing of quantitative information conveyed by symbolic and nonsymbolic stimuli led to the hypothesis that there exists a common cerebral representation of quantity (Dehaene, Dehaene-Lambertz, & Cohen, 1998). The neural basis underlying the encoding of number magnitude has been localized to regions in and around the intraparietal sulcus (IPS) by brain imaging studies. However, it has never been demonstrated that these same regions are also involved in the quantitative processing of nonsymbolic stimuli. Using functional brain imaging, we explicitly tested the hypothesis of a common substrate. Angles, lines, and two-digit numbers were presented pairwise, one to the left and one to the right of the fixation point. In the three comparison tasks, participants (n = 18) pressed the key on the side of the largest quantity. In the three control tasks, they indicated the side on which dimming occurred. A conjunction analysis between the three subtractions (comparison task-control task) revealed a site in left IPS that is specifically responsive when two stimuli have to be compared quantitatively, irrespective of stimulus format. The results confirm the hypothesis that quantity is represented by a common mechanism for both symbolic and nonsymbolic stimuli in IPS. In addition, the interaction between task and type of stimulus identified a region anterior to the conjunction site, not specific for quantitative processing, but reflecting general processes loaded by number processing. PMID- 12590843 TI - Reading speech from still and moving faces: the neural substrates of visible speech. AB - Speech is perceived both by ear and by eye. Unlike heard speech, some seen speech gestures can be captured in stilled image sequences. Previous studies have shown that in hearing people, natural time-varying silent seen speech can access the auditory cortex (left superior temporal regions). Using functional magnetic resonance imaging (fMRI), the present study explored the extent to which this circuitry was activated when seen speech was deprived of its time-varying characteristics. In the scanner, hearing participants were instructed to look for a prespecified visible speech target sequence ("voo" or "ahv") among other monosyllables. In one condition, the image sequence comprised a series of stilled key frames showing apical gestures (e.g., separate frames for "v" and "oo" [from the target] or "ee" and "m" [i.e., from nontarget syllables]). In the other condition, natural speech movement of the same overall segment duration was seen. In contrast to a baseline condition in which the letter "V" was superimposed on a resting face, stilled speech face images generated activation in posterior cortical regions associated with the perception of biological movement, despite the lack of apparent movement in the speech image sequence. Activation was also detected in traditional speech-processing regions including the left inferior frontal (Broca's) area, left superior temporal sulcus (STS), and left supramarginal gyrus (the dorsal aspect of Wernicke's area). Stilled speech sequences also generated activation in the ventral premotor cortex and anterior inferior parietal sulcus bilaterally. Moving faces generated significantly greater cortical activation than stilled face sequences, and in similar regions. However, a number of differences between stilled and moving speech were also observed. In the visual cortex, stilled faces generated relatively more activation in primary visual regions (V1/V2), while visual movement areas (V5/MT+) were activated to a greater extent by moving faces. Cortical regions activated more by naturally moving speaking faces included the auditory cortex (Brodmann's Areas 41/42; lateral parts of Heschl's gyrus) and the left STS and inferior frontal gyrus. Seen speech with normal time-varying characteristics appears to have preferential access to "purely" auditory processing regions specialized for language, possibly via acquired dynamic audiovisual integration mechanisms in STS. When seen speech lacks natural time-varying characteristics, access to speech-processing systems in the left temporal lobe may be achieved predominantly via action-based speech representations, realized in the ventral premotor cortex. PMID- 12590844 TI - Semantic processing in the left inferior prefrontal cortex: a combined functional magnetic resonance imaging and transcranial magnetic stimulation study. AB - The involvement of the left inferior prefrontal cortex (LIPC) in phonological processing is well established from both lesion-deficit studies with neurological patients and functional neuroimaging studies of normals. Its involvement in semantic processing, on the other hand, is less clear. Although many imaging studies have demonstrated LIPC activation during semantic tasks, this may be due to implicit phonological processing. This article presents two experiments investigating semantic functions in the LIPC. Results from a functional magnetic resonance imaging experiment demonstrated that both semantic and phonological processing activated a common set of areas within this region. In addition, there was a reliable increase in activation for semantic relative to phonological decisions in the anterior LIPC while the opposite comparison (phonological vs. semantic decisions) revealed an area of enhanced activation within the posterior LIPC. A second experiment used transcranial magnetic stimulation (TMS) to temporarily interfere with neural information processing in the anterior portion of the LIPC to determine whether this region was essential for normal semantic performance. Both repetitive and single pulse TMS significantly slowed subjects' reactions for the semantic but not for the perceptual control task. Our results clarify the functional anatomy of the LIPC by demonstrating that anterior and posterior regions contribute to both semantic and phonological processing, albeit to different extents. In addition, the findings go beyond simply establishing a correlation between semantic processing and activation in the LIPC and demonstrate that a transient disruption of processing selectively interfered with semantic processing. PMID- 12590845 TI - Common and segregated neuronal networks for different languages revealed using functional magnetic resonance adaptation. AB - The effect of word repetition within and across languages was studied in English Chinese bilinguals who read rapidly presented word pairs in a block design and an event-related fMRI study. Relatively less increase in MR signal was observed when the second word in a pair was identical in meaning to the first. This occurred in the English-only and mixed-languages conditions. Repetition-induced reductions in BOLD signal change were found in the left lateral prefrontal and lateral temporal regions in both types of conditions in the block experiment, suggesting that processing in these regions is sensitive to semantic features present in words and characters, and that part of the semantic neuronal networks serving English and Chinese is shared. In addition, these regions showed greater absolute signal change in the mixed-languages trials relative to the English-only trials. These findings were mostly replicated in an event-related experiment. Together, the experiments suggest that while the networks for Chinese and English word processing have shared components, there are also components that may be language specific. PMID- 12590846 TI - Repair, revision, and complexity in syntactic analysis: an electrophysiological differentiation. AB - One of the core aspects of human sentence processing is the ability to detect errors and to recover from erroneous analysis through revision of ambiguous sentences and repair of ungrammatical sentences. In the present study, we used event-related potentials (ERPs) to help identify the nature of these processes by directly comparing ERPs to complex ambiguous sentence structures with and without grammatical violations, and to simpler unambiguous sentence structures with and without grammatical violations. In ambiguous sentences, preference of syntactic analysis was manipulated such that in one condition, the structures agreed with the preferred analysis, and in another condition, a nonpreferred but syntactically correct analysis (garden path) was imposed. Nonpreferred ambiguous structures require revision, whereas ungrammatical structures require repair. We found that distinct ERPs reflected different characteristics of syntactic processing. Specifically, our results are consistent with the idea that a positivity with a posterior distribution across the scalp (posterior P600) is an index of syntactic processing difficulty, including repair and revision, and that a frontally distributed positivity (frontal P600) is related to ambiguity resolution and/or to an increase in discourse level complexity. PMID- 12590848 TI - Long-latency ERPs and recognition of facial identity. AB - N400 brain event-related potential (ERP) is a mismatch negativity originally found in response to semantic incongruences of a linguistic nature and is used paradigmatically to investigate memory organization in various domains of information, including that of faces. In the present study, we analyzed different mismatch negativities evoked in N400-like paradigms related to recognition of newly learned faces with or without associated verbal information. ERPs were compared in the following conditions: (1) mismatching features (eyes-eyebrows) using a facial context corresponding to the faces learned without associated verbal information ("pure" intradomain facial processing); (2) mismatching features using a facial context corresponding to the faces learned with associated occupations and proper names ("nonpure" intradomain facial processing); (3) mismatching occupations using a facial context (cross-domain processing); and (4) mismatching names using an occupation context (intradomain verbal processing). Results revealed that mismatching stimuli in the four conditions elicited a mismatch negativity analogous to N400 but with different timing and topographical patterns. The onset of the mismatch negativity occurred earliest in Conditions 1 and 2, followed by Condition 4, and latest in Condition 3. The negativity had the shortest duration in Task 1 and the longest duration in Task 3. Bilateral parietal activity was confirmed in all conditions, in addition to a predominant right posterior temporal localization in Condition 1, a predominant right frontal localization in Condition 2, an occipital localization in Condition 3, and a more widely distributed (although with posterior predominance) localization in Condition 4. These results support the existence of multiple N400, and particularly of a nonlinguistic N400 related to purely visual information, which can be evoked by facial structure processing in the absence of verbal-semantic information. PMID- 12590847 TI - Time course of processes and representations supporting visual object identification and memory. AB - Event-related potentials (ERPs) were used to delineate the time course of activation of the processes and representations supporting visual object identification and memory. Following K. Srinivas (1993), 66 young people named objects in canonical or unusual views during study and an indirect memory test. Test views were the same or different from those at study. The first ERP repetition effect and earliest ERP format effect started at approximately 150 msec. Multiple ERP repetition effects appeared over time. All but the latest ones were largest for same views, although other aspects of their form specificity varied. Initial ERP format effects support multiple-views-plus-transformation accounts of identification and indicate the timing of processes of object model selection (frontal N350 from 148-250 to 500-700 msec) and view transformation via mental rotation (posterior N400/P600 from 250-356 to 700 msec). Thereafter, a late slow wave reflects a memory process more strongly recruited by different than same views. Overall, the ERP data demonstrate the activation of multiple memory processes over time during an indirect test, with earlier ones (within 148 400 msec) characterized by a pattern of form specificity consistent with the specific identification-related neural process or representational system supporting each memory function. PMID- 12590849 TI - Nasal polyps: still more questions than answers. AB - Nasal polyps are common, affecting one to four per cent of the population. Their cause, however, remains unknown and it is possible that it is not the same in all patients. They have a clear association with asthma, aspirin sensitivity and cystic fibrosis. Histologically they demonstrate large quantities of extracellular fluid, mast cell degranulation and an infiltrate of inflammatory cells, usually eosinophils. While this appearance would suggest an allergic pathology there is little conclusive evidence to support this in most patients. There is, however, some preliminary evidence to suggest that a local allergic process could be the cause. While allergic fungal sinusitis is a well defined clinical entity with recognized diagnostic criteria the ubiquitous nature of fungal spores makes the role of fungal infection in patients with nasal polyps difficult to determine and currently this remains unclear. Surgical treatment of nasal polyps has declined in recent years as the benefits of medical treatment have become increasingly recognized. There is good evidence to support the use of corticosteroids both as a primary and post-operative treatment in the majority of patients. Other medical treatments require further evaluation before they could be considered a viable alternative to steroids. Assessment of the literature regarding surgical intervention is difficult and there is little evidence on which to base a surgical treatment philosophy. The authors believe that an endoscopic approach using a microdebrider facilitates accurate removal of polyps with preservation of normal anatomy. PMID- 12590850 TI - Treatment of Meniere's disease by intratympanic gentamicin application. AB - Meniere's disease is a vestibular disorder characterized by episodic vertigo, tinnitus, fluctuant hearing loss, and ear fullness, with vertiginous attacks being the most disabling complaint. The effectiveness of conservative treatment has been variable, while surgical techniques provide more permanent relief from vertigo, but pose possible morbidity and cochlear risk. Intratympanic administration of gentamicin has been proposed as an alternative for patients with debilitating Meniere's disease who have failed to respond to standard medical treatment. The goal of such treatment is to control vertigo by partially or completely destroying the vestibular system, while at the same time maintaining hearing. In this review we present the current medical literature on pathophysiology, pharmacokinetics, administration methods, dosage, treatment protocols, and problems related to intratympanic administration of gentamicin for patients with MD. Intratympanic gentamicin administration is a simple medical modality for treatment of persistent vertigo in patients with Meniere's disease. According to recent world research protocols, we propose the instillation of gentamicin by transtympanic injection, as a quick, easy, well-tolerated, ambulatory and cost-effective technique. Drug solution concentrations should be dependent on the frequency, intensity and duration of vertigo spells, as well as the degree of existing hearing loss, thus providing progressive vertigo relief with a low possibility of secondary deafness. PMID- 12590851 TI - Its pages are numbered but are its days? PMID- 12590852 TI - Philipp Friedrich Arnold, Ludvig Levin Jacobson and their contribution to head and neck anatomy. AB - Philipp Friedrich Arnold and Ludvig Levin Jacobson were two of the greatest anatomists of the 19th century. Within this paper their backgrounds and their contribution to the development of various aspects of head and neck anatomy are considered. PMID- 12590853 TI - External auditory canal duplication anomalies associated with congenital aural atresia. AB - Maldevelopment of the first branchial cleft can produce a broad spectrum of anomalies in its derivative structure, the external auditory canal (EAC). Failure of the cleft to develop normally can result in either the absence of a normally patent EAC (atresia, or stenosis) or a duplication anomaly (cyst, sinus, or fistula). Despite their common origins, the coexistence of these anatomical abnormalities is quite unusual. We present four patients with both aural atresia and duplication anomalies of the EAC. Three patients had non-syndromic unilateral aural atresia and presented with periauricular lesions originating from the first branchial cleft. The other patient had a variant of Treacher Collins syndrome and presented with draining infra-auricular fistulae. The classification and management of first branchial cleft anomalies is reviewed in light of these cases. An understanding of the embryogenesis of the external ear is necessary to successfully recognize and treat this spectrum of deformities. A classification system is presented that encompasses the full spectrum of first cleft anomalies. PMID- 12590854 TI - Otomycosis in Turkey: predisposing factors, aetiology and therapy. AB - Otomycosis usually requires long-term treatment and tends to recur. This study was performed on 87 patients with the clinical diagnosis of otomycosis and 20 controls in order to determine the pathogenic agents, predisposing factors and a cost-effective treatment. The predisposing factors included wearing head clothes (74.7 per cent), presence of dermatomycoses (34.5 per cent) and swimming (27.6 per cent). The most common pathogenic fungus was Aspergillus niger (44.8 per cent) in the otomycosis group. The only isolate was Candida albicans in the control group (2.5 per cent). We concluded that administration of four per cent boric acid solution in alcohol and frequent suction cleaning of the ear canal might be a cost-effective treatment for otomycosis since 77 per cent of the patients were treated effectively this way. Eighty per cent of the resistant cases had mixed fungal-bacterial infections, and 50 per cent of them had dermatomycoses. These resistant cases were treated by administration of tioconazole ointment. PMID- 12590855 TI - Comparison of the efficacy and safety of moxifloxacin and trovafloxacin for the treatment of acute, bacterial maxillary sinusitis in adults. AB - In this multicentre, multinational, comparative, double-blind clinical trial, out patients with both symptoms and radiographic evidence of acute sinusitis were randomly assigned to receive either a seven-day, once daily (o.d.) oral regimen of moxifloxacin (400 mg) or a 10-day o.d. oral regimen of trovafloxacin (200 mg). Among 452 patients considered valid for clinical efficacy, moxifloxacin treatment was found to be statistically equivalent to trovafloxacin (96.9 per cent vs 92.1 per cent -95 per cent CI = 0.6 per cent; 8.9 per cent) at the seven to 10 days post-therapy assessment. At follow-up, the success rate in the moxifloxacin group was 94.9 per cent and that for the trovafloxacin group was 97.6 per cent (95 per cent CI = -4.9 per cent; 1.3 per cent). The predominant causative organisms were Streptococcus pneumoniae, Haemophilus influenzae and Staphylococcus aureus followed by Enterobacteriaceae and Moraxella catarrhalis. The bacteriological success rate at the post-therapy evaluation was similar in both treatment groups: 94.4 per cent and 90.1 per cent in the moxifloxacin and trovafloxacin groups respectively (95 per cent CI = -3.0 per cent; 11.9 per cent). Only three of the 103 baseline isolated pathogens still persisted in the moxifloxacin group, whereas there were 10 of the 121 isolates that failed to respond in the trovafloxacin treatment group. At least one drug-related event was reported by 16.9 per cent of the moxifloxacin-treated patients and by 22.3 per cent of those who received trovafloxacin. CNS events such as dizziness and vertigo were reported more than five times more often in patients receiving trovafloxacin than in the moxifloxacin group. Trovafloxacin recipients were also more than twice as likely to discontinue treatment due to adverse events than moxifloxacin-treated patients. Overall, moxifloxacin was at least as effective clinically and bacteriologically as trovafloxacin and better tolerated. PMID- 12590856 TI - Nasal septal perforation repair using open septoplasty and unilateral bipedicled flaps. AB - The aim of this study was to assess the success of the technique of open septoplasty with bipedicled flaps in achieving long-term closure and control of symptoms. Between 1993 and 2000, 32 patients underwent surgery for nasal septum perforation. Twenty patients with large perforations underwent posterior edge repair only. The remaining 12 patients (five female, and seven male) with perforations less than 20 mm in diameter underwent open septoplasty and a unilateral bipedicled flap closure. A retrospective review involving a symptom scores assessment and follow-up examination was conducted. One patient died of unrelated illness, and was excluded. Results showed 10 of the remaining 11 patients achieved closure after a mean follow up of 10 months. The symptoms of crusting (p < 0.0001), epistaxis (p < 0.02), discharge (p < 0.012), whistling (p < 0.011), and overall discomfort (p < 0.02), were all significantly improved. We conclude that by using this technique on patients with small perforations less than 20 mm, it is possible to achieve up to a 90 per cent perforation closure rate, and a significant improvement in patients' symptoms. PMID- 12590857 TI - Voice rehabilitation after laryngectomy with the Provox voice prosthesis in South Africa. AB - A study was undertaken to determine whether the Provox voice prosthesis provides good voice rehabilitation following a total laryngectomy in the urban, suburban and rural populations served by a tertiary referral hospital in South Africa. Between 1995 and 1999, a cohort of 128 patients at Tygerberg Hospital was rehabilitated with the Provox voice prosthesis after laryngectomy. In 104 patients primary placement of the prosthesis was done at the time of the laryngectomy. Mean device life and adverse events were determined. Voice quality was assessed subjectively in 104 patients and objectively in 26 patients. The mean device life was 303 days and adverse events occurred in 16 patients. Subjectively, 77 of 104 patients had a good voice, and objectively 22 of 26 patients had good voice intelligibility. The Provox voice prosthesis provides good voice rehabilitation following total laryngectomy, with minimal complications, in the population served by Tygerberg Hospital. PMID- 12590858 TI - The use of the KTP laser in the management of hypertrophic tracheal mucosa and granulation tissue around Provox valve protheses. AB - The development of hypertrophic mucosa and granulation tissue around a tracheo oesophageal fistula voice prosthesis is a commonly encountered problem occurring in 15-20 per cent of patients. Upsizing of the voice prosthesis as well as cautery (silver nitrate and electocautery) are used to counteract this problem to a varying degree of success. The use of the CO2 laser has been mentioned by some authors but details of the method and results have not been published. We report the successful use of the KTP laser for removing such granulations whilst retaining the valve and discuss the reasons why this method should be considered. PMID- 12590860 TI - Simple, painless, cosmetic closure of endaural incisions. AB - Endaural incisions provide a cosmetic approach to the middle-ear cleft, tympanic membrane for myringoplasty, and attic and antrum for small cavity mastoidectomy. These wounds have traditionally been closed with sutures, but platelet or fibrin adhesives provide an alternative. However, discomfort at the time of suture removal, particularly in children, and the risk of transmission of blood-borne infections in gels are limitations to these wound closure techniques. This paper describes our experience with Histoacryl glue (butyl-2-cyanoacrylate) in closing endaural incisions. PMID- 12590859 TI - Management of N0 neck in laryngeal carcinoma. Impact on patient's survival. AB - Management of patients with carcinoma of the larynx should systematically include an appropriate treatment of lymph nodes according to the TNM stage. One of the most controversial points of the treatment in these patients is the management of the clinically negative neck (N(0)). A retrospective study of 295 patients with laryngeal carcinoma and N(0) neck undergoing treatment in our centre between 1983 and 1993 is presented. We observed a significant decrease in the survival of clinically N(0) patients with histologically affected lymph nodes. Lymphadenopathy was more frequently detected in patients with supraglottic tumours (38 per cent) when compared to glottic tumours (16 per cent). In our experience, routine bilateral and unilateral dissection of N(0) necks in all supraglottic tumours and in T3-T4 glottic tumours, respectively, is the most beneficial approach for patients in terms of survival. PMID- 12590861 TI - Necrotizing fasciitis: a rare occurrence in the external auditory meatus. AB - The diagnosis and management of a patient with a sloughing otitis externa thought to be necrotizing fasciitis is described. The clinical characteristics of this condition, diagnostic difficulties faced and a brief literature review are presented. PMID- 12590862 TI - Gunshot injuries to the temporal bone. AB - The current incidence of missile injury to the temporal bone (MITB) is very low in the United Kingdom. However, the increasing frequency of firearm violence in Britain suggests a greater risk of occurrence. This, along with the devastating potential sequelae of MITB (facial palsy, dead ear, intracranial damage, major vascular injury and cosmetic disfigurement), requires otolaryngologists to be conversant with all aspects of their management. The risk of major complications is much higher with MITB than with temporal bone injury following blunt trauma, and surgical management is, therefore, much more common. We present one such case, and review the literature outlining the pathogenesis, clinical features, and recommended management. PMID- 12590863 TI - Epithelioid haemangioendothelioma arising in the nasal cavity. AB - We report here the case of an epithelioid haemangioendothelioma (EHE) arising in the nasal cavity which is, to the best of our knowledge, the first ever described example in the world literature in that particular site. The patient is a 23-year old male who presented with repeated episodes of epistaxis from the nasal cavity and with a 1.5 cm reddish, polypoid, smooth, spontaneously bleeding nodule in the right middle meatus. This lesion was histologically diagnosed as epithelioid haemangioendothelioma. Immunohistochemically the neoplasm displayed striking positivity for CD31, CD34 and vimentin. A surgical approach was performed by 'facial degloving', removing the right inferior turbinate, the anterior two thirds of the middle turbinate and the medial wall of the ethmoid bone. After 12 months follow-up the patient is disease-free, without any local or distant recurrence. PMID- 12590864 TI - Use of a nitinol stent to treat dysphagia post-laryngopharyngectomy. AB - Permanent dysphagia occurring after laryngectomy or laryngopharyngectomy is uncommon, and when it does occur, can usually be treated by periodic dilatation under general anaesthesia. Occasionally, however, conservative treatment is insufficient, and patients require long-term feeding via a gastrostomy or jejunostomy tube. We describe the case of a man with an anastamotic stricture post-pharyngectomy who underwent insertion of a nitinol stent across the stricture for treatment of dysphagia. The patient's swallowing was significantly improved in the short-term, however, ultimately, florid granulation tissue formation led to obstruction of the stent and a disappointing long-term result. This, to our knowledge, is the first documentation of the use of a stent to treat dysphagia in a patient post-laryngectomy or pharyngolaryngectomy. PMID- 12590865 TI - Tissue augmentation using Bioplastique as a treatment of leakage around a Provox 2 voice prosthesis. AB - Rehabilitation of voice and speech after laryngectomy with valve prosthesis has become a well-established practice in recent years. The formation of tracheo oesophageal fistula (TOF) and the subsequent management of the patient with a voice prosthesis can be associated with a number of problems and complications. We report a new technique of the use of injectable Bioplastique in the treatment of persistent leakage around Provox 2 voice prosthesis. Our experience in two cases has shown that it is a relatively simple and effective procedure in stopping the leak around the valve immediately and is without any short-term complications. PMID- 12590866 TI - Polyflex stent: is it radiopaque enough? AB - We report a case of tracheal stenosis in which a migrated Polyflex (Rusch AG, Germany) stent entered the right main bronchus but could not be identified on the chest radiograph. The stent was identified at bronchoscopy and removed under general anaesthesia. Polyflex stents are radiopaque but may not always show up on radiograph. We recommend reliance on clinical symptoms rather than imaging to diagnose migration. If stent migration is suspected then imaging should include thoracic inlet films that incorporate a lateral view. PMID- 12590868 TI - Cardiovascular effects of an immunosuppressive agent cyclosporin A. AB - Cyclosporin A (CsA) is now routinely used for transplantation of all solid organs, bone marrow transplantation, and for an increasing number of immunological diseases. However, treatment with CsA is an important iatrogenic cause of post-transplant hypertension, hyperlipidemia, and diabetes, which may contribute to the high cardiovascular morbidity in transplant recipients. Recently, the calcineurin inhibitor CsA has been employed in vivo and in vitro to examine the role of calcineurin in the signal transduction leading to cardiac hypertrophy. A cell culture study demonstrated the inhibitory effect of CsA on cytokine production by cardiac myxoma cells, the most common primary tumor of the heart. This review discusses recent data on the cardiovascular effects of CsA. PMID- 12590867 TI - An unusual metastatic site of laryngeal carcinoma: scapular muscles. AB - It is well known that in advanced cervical metastatic disease of head and neck squamous cell carcinoma, the incidence of distant metastasis is high. A case of distant metastasis to the scapular muscles from an uncontrolled cervical metastasis of laryngeal carcinoma is presented. Other unusual metastatic sites are reviewed and the possible mechanism of the spread is discussed. PMID- 12590869 TI - Cyclosporin A-induced autoimmunity in the rat: central versus peripheral tollerance. AB - No abstract available PMID- 12590870 TI - Topical cyclosporin in the treatment of dermatologic diseases. AB - Cyclosporine A (CsA) has been in clinical use for some decades, primarily for the prevention and treatment of organ transplant rejection and graft-versus-host disease. In more recent years, Cyclosporine has been recognized as beneficial in the treatment of dermatologic diseases, such as: psoriasis, lichen planus, Behcet disease, atopic dermatitis, pyoderma gangrenosum and epidermolysis bullosa acquisita. Above all, Cyclosporine is an important therapeutic modality for several dermatologic diseases that are refractory to other agents. PMID- 12590871 TI - Biological effects of electromagnetic fields. AB - Nowadays, concerns about hazards from electromagnetic fields represent an alarming source for human lives in technologically developed countries. We are surrounded by electromagnetic fields everywhere we spend our working hours, rest or recreational activities. The aim of this review is to summarize the biological effects due to these fields arising from power and transmission lines, electrical cable splices, electronic devices inside our homes and work-places, distribution networks and associated devices such as cellular telephones and wireless communication tower, etc. Special care has been reserved to study the biological effects of electromagnetic fields on cell lines of the mammalian immune system about which our research group has been working for several years. PMID- 12590872 TI - Monocyte chemotactic protein-1 in a randomized placebo controlled study of canine plasmacytic-lymphocytic colitis. AB - The purpose of this study was to determine serum and colonic monocyte chemotactic protein-1 (MCP-1) concentration in dogs with plasmacytic-lymphocytic (PL) colitis, as well as to demonstrate if the concentration of MCP-1 may be an accurate diagnostic and prognostic marker for PL colitis in dogs receiving three different therapeutic protocols. Serum and colonic MCP-1 concentration were measured in 18 dogs with PL colitis and in 6 controls. Dogs with PL colitis were randomly divided in 3 groups and for a period of 30 days received the following: Group 1, sulfasalazine, prednisone and placebo; Group 2, placebo and 㲓/㲖 fatty acids, and Group 3, sul-fasalazine, prednisone and 㲓/㲖 fatty acids. Colonic and serum MCP-1 concentration were de termined at the beginning and at the end of this period. Serum MCP-1 concentrations at the be-ginning were not significantly increased in dogs with PL colitis compared to the controls. On the contrary, colonic tissue MCP-1 concentrations at the beginning were significantly increased in the same dogs compared to the controls. Moreover, the colonic tissue MCP-1 concentration in the dogs of group 3 was significantly decreased (Pπ.001) at the end of the experiment. The same observations were obtained from dogs of groups 1 and 2, in which the colonic MCP-1 concentra-tion also significantly decreased (Pπ.01). These data demonstrate that in canine PL colitis, colo-nic MCP-1 concentration is increased in comparison to those of controls and suggest that the colonic MCP-1 concentration may aid in the diagnosis of canine PL colitis. PMID- 12590873 TI - Cetirizine, a second-generation H1 antagonist, modulates Rantes and MCP-1 levels in allergic rhinitis. AB - Chemokines and their receptors are involved in several allergic diseases. We measured RANTES and MCP-1 levels in sera of allergic rhinitis patients, and also we evaluated the effect of cetrizine, a second-generation H1 antagonist, on these chemoattractant proteins. 15 subjects were studied (10 males and 5 females; mean age: 26.7 years). They were suffering from perennial or seasonal allergic rhinitis induced by Dermatophagoides pteronyssinus (8 patients) or by grasses (7 patients). RANTES and MCP-1 serum levels were detected with an enzyme immunoassay before and after two weeks of treatment with 10 mg of cetirizine daily, and again after two weeks of washout. Baseline serum levels of RANTES and MCP-1 chemokines were significantly higher (p < 0.02 and p = 0.007, respectively) in allergic patients than in the healthy control group. Cetirizine resulted in a significant decrease in RANTES (p < 0.02) and MCP-1 (p = 0.003) versus baseline values. There is an increase in RANTES and MCP-1 in allergic rhinitis, which is counteracted by cetirizine. PMID- 12590874 TI - Serum procalcitonin, adenosine deaminase and its isoenzymes in the aetiological diagnosis of pneumonia in children. AB - A prospective study was undertaken to assess the usefulness of leukocyte count, serum C-reactive protein (CRP), procalcitonin (PCT), and the activities of total adenosine deaminase (tADA) and its isoenzymes ADA1 and ADA2, in the aetiological diagnosis of pneumonia in children. The study included three groups. Group A consisted of 23 children with bacterial pneumonia, group B of 50 children with viral and mycoplasmal pneumonia and group C of 46 healthy children. On the first day of admission in the clinic, blood samples were collected before the start of antimicrobial treatment, for culture, serological tests, leukocyte count and for the determination of CRP and PCT levels as well as tADA activity and its isoenzymes ADA1 and ADA2. According to our results, the mean leukocyte count and the mean concentrations of PCT and CRP were significantly higher in the children of group A than those in groups B and C. The admission serum PCT concentration has a higher sensitivity, specificity and positive predictive value for bacterial pneumonia than either CRP or the leukocyte count. The mean serum tADA, ADA1 and ADA2 activity in children of group A was not significantly different from those in group C, while the difference between groups B and C was statistically significant. In conclusion, we found that CRP is a good marker for screening various infectious diseases, but it cannot be used to distinguish between bacterial and viral infections. Serum PCT measurement might be a useful tool for the physician for the aetiological diagnosis of pneumonia in children. Measurements of serum tADA and ADA2 activity may provide useful additional diagnostic information on the aetiology of pneumonia so that appropriate antibiotic therapy can be given promptly. Further studies with larger patients groups are required to confirm our results. PMID- 12590875 TI - Reverse transcriptase mutations in HIV-1 infected patients treated with two nucleoside analogues: the SMART study. AB - Resistance to nucleoside reverse transcriptase inhibitors (NRTIs) was studied in 527 HIV-1-infected patients, 342 responder and 185 non-responder to two NRTIs. Responders were followed for one year to assess the incidence of clinical failure. The prevalence of the 215Y/F substitution was higher among non responder, compared to responder patients (33.7% vs. 17%, P = 0.0005), whereas the prevalence of the 184V and of the 70R mutations was comparable between these two groups. The 74V substitution was never observed and the 75T mutation was detected in only two subjects non-responder to a stavudine including regimen. Reduced susceptibility to didanosine or stavudine was infrequent. Reduced susceptibility to zidovudine was observed in 25% of individuals failing a zidovudine including regimen, whereas reduced susceptibility to lamivudine was detected in all subjects failing a lamivudine including regimen. In the prospective analysis, patients with undetectable viral load at enrollment had a lower incidence of failure rate over one year compared to those with detectable HIV-RNA at entry (P < 0.0001). A detectable viral load at enrollment was the only independent variable that predicted clinical failure over one year (P < 0.0001). PMID- 12590876 TI - Nasal immunotherapy is effective in the treatment of rhinitis due to mite allergy. A double-blind, placebo-controlled study with rhinological evaluation. AB - The aim of this paper is to evaluate the efficacy of intranasal hyposensitizing therapy in perennial rhinitis. 36 patients suffering from perennial allergic rhinitis (Dermatophagoides-sensitive) underwent a double blind placebo-controlled trial for a period of 8 months. The efficacy of nasal immunotherapy was evaluated by collecting symptoms score and evaluating objective rhinological parameters (nasal resistance, cross areas and volumes, mucociliary clearance times, specific nasal provocation threshold). A significant improvement (pπ,01) of symptom score of active against placebo group was observed after treatment. Also objective nasal parameters (total nasal resistances, mucociliary clearance, C notch area, and provocative threshold) significantly (pπ,01) improved after treatment. Adverse local reactions were rare and did not interfere with the protocol. The results underline the efficacy and quickness of local nasal immunotherapy in the treatment of perennial allergic rhinitis documented by the improvement of subjective and objective parameters. PMID- 12590877 TI - Effectiveness of montelukast versus budesonide on quality of life and bronchial reactivity in subjects with mild-persistent asthma. AB - Insufficient data exist to evaluate the comparative effects of inhaled corticosteroids (ICS) versus leukotriene receptor antagonist (LTRA) on airway inflammation and quality of life (QoL). The aim of the study was to compare the effectiveness of montelukast compared to budesonide at different doses on QoL and bronchial reactivity in mild-asthmatic adult patients. 45 subjects with bronchial asthma were randomly assigned to a different treatment and divided in 3 treatment groups: A: 400 mg of budesonide twice a day; B: 10 mg of montelukast daily; C: 10 mg of montelukast daily plus 400 mg of budesonide twice a day. At the beginning of the study and at the end of the treatment period (16 weeks) all patients underwent complete clinical evaluation, pulmonary function testing and methacholine challenge test (MCHt). In group A the increase from baseline was 153.4%, in group C was 133.2%, and in group B 247.7%, the latter increase being statistically significant compared to that in the other 2 groups (p< 0.005 Wilcoxon test). In all domains the improvement in quality of life in the group treated with montelukast (group B) was significantly greater than that in the group treated with both medications (group C): in particular, the improvement was consistent in the symptoms (p< 0.01) and emotions (p< 0.01) domains, and weaker in the physical activity (p< 0.05). A similar difference was observed between group B and A, but only in the symptoms (p<0.01), emotions (p<0.01), and environmental stimuli domains (p<0.05). The personal perception of their own disease is important for a correct therapeutic management of asthma. In order to optimize the treatment, a complete adherence of the patient to the treatment itself is required, to be achieved through simplification of therapeutic schedule and easy administration of medications. Montelukast may be considered a valid alternative in the treatment of mild-persistent asthma, both for the clinical and functional benefits and for the great advantage of the once-daily dosage, which consistently improves the compliance with the chronic treatment of the disease. PMID- 12590879 TI - Status of hyaluronan supplementation therapy in osteoarthritis. AB - Hyaluronans are polysaccharide molecules that occur naturally in synovial fluid; they help to create a viscous environment, cushion joints, and maintain normal function. The American College of Rheumatology recommends intra-articular injection of hyaluronans, which are available as several distinct therapeutic products, as an alternative to oral analgesics or nonsteroidal anti-inflammatory drugs for the symptomatic treatment of pain associated with osteoarthritis of the knee. A large body of literature supports the clinical efficacy and safety of this therapeutic class for this indication, although there are differences between the marketed products and they should be evaluated independently. Preliminary work investigating the use of hyaluronans for osteoarthritis in joints other than the knee has also produced promising results. There is growing evidence that hyaluronans, the biology of which is complex, may also have structure-modifying activity. Thus, compared with currently approved nonoperative therapies for osteoarthritis, hyaluronans may also have beneficial effects on the disease process in osteoarthritis. PMID- 12590880 TI - The role of bone metabolism in osteoarthritis. AB - Bone is not structurally normal in osteoarthritis (OA). Periarticular bone in OA has increased turnover, decreased bone mineral content and stiffness, and decreased trabecular numbers. The increased turnover of collagen, in particular, may affect the biomechanical competence of bone and is driven by a number of processes, including osteoblast phenotypic expression, chondrocyte apoptosis, matrix metalloproteinases, and growth factors. This turnover is reflected in alterations in biomarkers and bone mineral density (BMD). High BMD at non-joint sites is associated with an increased risk of OA; however, low BMD and high bone turnover appears to be associated with more rapid progression. These alterations in bone turnover offer new therapeutic opportunities. PMID- 12590882 TI - Imaging and osteoarthritis: what is the predictive value? AB - Recent protocols for standardized knee radiography, which attempts to image the knee with reproducible, parallel alignment of the medial tibial plateau and radiograph beam, possess many theoretic advantages. As a group, they permit measurement of tibiofemoral joint space width with remarkable precision--the sine qua non of sensitive detection of change. However, only limited longitudinal data are available in peer-reviewed publications to permit a direct evaluation of the suitability of these protocols for use in multicenter clinical trials and studies of biomarkers of osteoarthritis (OA) progression. Longitudinal data from several National Institutes of Health-supported studies of OA progression, as reflected in radiographs acquired with high levels of standardization for radioanatomic positioning of the knee, should be available in the next several years. Alternatively, data from the placebo groups of several industry-supported phase III trials of purported disease-modifying OA drugs, which were terminated prematurely because of adverse events or lack of efficacy, may be made available for rapid analysis regarding the performance of current standardization protocols with respect to their sensitivity to disease progression. PMID- 12590881 TI - Joint irrigation as treatment for osteoarthritis. AB - Irrigation is necessary to achieve distension and visualization of the joint during arthroscopy. Clinical improvement observed after diagnostic arthroscopy has been attributed to irrigation, and this is supported by sound pathophysiologic rationales. Studies have been performed using various irrigation techniques with and without concomitant arthroscopy. Unblinded studies have generally yielded favorable results, although recent blinded studies have demonstrated a substantial and durable placebo effect, but little support for the benefit of the irrigation procedure. PMID- 12590884 TI - Estrogen therapy in postmenopausal women. PMID- 12590883 TI - Advanced glycation endproducts and osteoarthritis. AB - Non-enzymatic glycation of proteins, such as collagen, results in the formation of advanced glycation endproducts (AGE). Advanced glycation endproducts result in pathologic stiffening of cartilage and extracellular matrix and accumulate with age. Pentosidine, an AGE, is present in serum, synovial fluid, and articular cartilage from patients with osteoarthritis (OA). However, AGE levels are not always increased, and may be decreased locally, in association with osteoarthritic pathology. The finding of pentosidine in articular cartilage of individuals with OA may not be specific for that disease, independent of chronologic age. Advanced glycation endproduct modification of normal articular cartilage increases its stiffness, increases chondrocyte-mediated proteoglycan degradation, reduces its susceptibility to matrix metalloproteinase-mediated degradation, and decreases proteoglycan synthesis by chondrocytes. These observations parallel findings in osteoarthritic cartilage, which suggests that AGE modification could contribute to the pathogenesis of OA. However, a causative link between AGEs and OA has not yet been established. PMID- 12590885 TI - How to grow bone to treat osteoporosis and mend fractures. AB - There is a need for "anabolic" drugs that can directly stimulate bone growth, improve bone microarchitecture, accelerate fracture healing, and, thus, restore bone strength to osteoporosis patients and, hopefully, regenerate eroded bone in arthritis patients. The anabolic agents currently leading the way to the clinic are the parathyroid hormone (PTH) and some of its adenylyl cyclase-stimulating fragments. This article is a summary of what is known about how PTHs stimulate bone growth. The controversial bone anabolic activities of the cholesterol lowering lipophilic statins are also described, and mechanisms by which they may stimulate bone growth are presented. Finally, evidence is presented for the body's "fat-o-stat" cytokine--leptin--indirectly restraining bone growth via a hypothalamic factor, while at the same time serving as a local PTH-like autocrine/paracrine stimulator of osteoblast activity, as well as an inhibitor of osteoclast generation. PMID- 12590886 TI - Identification and treatment of osteoporosis in fractures. AB - Patients who present with osteoporotic fracture are at highest risk of further fractures and their associated morbidity. Despite the availability of several evidence-based therapeutic options, which have the potential to reduce the incidence of fractures by up to 50%, it is paradoxical that these high-risk patients are seldom assessed for osteoporosis and offered treatment. Secondary prevention of osteoporotic fractures should now be the priority for osteoporosis services; the challenge that remains is to devise new models of patient care that can deliver strategies for the secondary prevention of osteoporotic fractures in different healthcare settings. PMID- 12590889 TI - Should relatives be denied access to the resuscitation room? AB - Within a health care system that promotes choice and autonomy, it no longer seems appropriate to exclude relatives from the resuscitation room. There is a growing body of research that suggests there are indeed many long-term benefits to be gained from witnessing the resuscitation of a loved one. There seems no doubt that relatives would like the opportunity to spend the last few valuable minutes with their loved one to say goodbye. However, it is the views of many staff working in the critical care setting that appear to be preventing witnessed resuscitation from becoming normal practice. This paper considers the staffs', the relatives' and the patients' perspectives on witnessed resuscitation and concludes that the majority of relatives should not be denied access to the resuscitation room. PMID- 12590888 TI - Determinants of fracture risk in osteoporosis. AB - Osteoporosis is a major public health issue, with fragility fractures of the hip, vertebrae, and distal radius considered the most important consequences. These lead to increased morbidity, mortality, hospital care, and dependency. The risk factors for the development of fragility fractures are numerous and involve genetic and environmental influences, as well as an interaction between the two. In this review, the recent literature examining genetic factors, possible candidate genes, the evolving area of intrauterine fetal programming, and anthropometric and environmental factors will be reviewed. PMID- 12590887 TI - Bisphosphonate mechanism of action. AB - The nitrogen-containing bisphosphonates (N-BPs), alendronate and risedronate, are the only pharmacologic agents shown to prevent spine and nonvertebral fractures associated with postmenopausal and glucocorticoid-induced osteoporosis. At the tissue level, this is achieved through osteoclast inhibition, which leads to reduced bone turnover, increased bone mass, and improved mineralization. The molecular targets of bisphosphonates (BPs) have recently been identified. This review will discuss the mechanism of action of BPs, focusing on alendronate and risedronate, which are the two agents most widely studied. They act on the cholesterol biosynthesis pathway enzyme, farnesyl diphosphate synthase. By inhibiting this enzyme in the osteoclast, they interfere with geranylgeranylation (attachment of the lipid to regulatory proteins), which causes osteoclast inactivation. This mechanism is responsible for N-BP suppression of osteoclastic bone resorption and reduction of bone turnover, which leads to fracture prevention. PMID- 12590890 TI - A randomised controlled trial comparing the use of manual versus mechanical compression to obtain haemostasis following coronary angiography. AB - Cardiac interventions have become a commonly accepted treatment option for patients with coronary heart disease. Managing the arterial puncture site and femoral sheath removal is an important aspect of cardiac nursing practice for patients who have had cardiac diagnostic and interventional procedures. The purpose of this study was to compare the use of manual compression with a mechanical compression device in achieving haemostasis after femoral sheath removal in coronary angiography patients and to determine the ability of these two techniques to reduce groin complications. A randomised controlled trial comparing two compression protocols (manual and QuicKlamp) was undertaken on a sample of 100 patients scheduled to have coronary angiography. Descriptive statistics were used to analyse and describe the data. Inter-group comparisons were analysed using either Chi-squared analysis for nominal data, or the Mann Whitney U-test for continuous variables. The results indicated that the QuicKlamp device took longer to effect haemostasis after femoral sheath removal (P=0.000) and subjects took longer to mobilise than after manual compression (P=0.001). More haematomas occurred following manual compression after pressure dressing removal (P=0.027). At 5-day follow-up, more bruising was identified in those subjects in the QuicKlamp compression group (P=0.046), as was swelling in female subjects (P=0.044). More episodes of chest pain at 5-day follow-up were identified in the manual compression group (P=0.014). The findings demonstrate that QuicKlamp mechanical compression is a safe alternative to manual compression for attaining haemostasis after femoral sheath removal. PMID- 12590891 TI - Music therapy--a complementary treatment for mechanically ventilated intensive care patients. AB - The aim of this study was to ascertain whether music therapy had a measurable relaxing effect on patients who were temporarily on a respirator in an intensive care unit (ICU) and after completion of respirator treatment investigate those patients' experiences of the music therapy. In the study both quantitative and qualitative measurements were applied. Twenty patients were included using consecutive selection. It became apparent that the patients remembered very little of their time in ICU. The analysis of the quantitative data showed a significant fall in systolic and diastolic blood pressure during the music therapy session and a corresponding rise after cessation of treatment. All changes were found to be statistically significant. The conclusion was that intensive care nursing staff can beneficially apply music therapy as a non pharmacological intervention. PMID- 12590892 TI - Autonomy among intensive care nurses in Finland. AB - The purpose of this study was to describe Finnish ICU nurses' (n=172) experiences of autonomy in their work. The research was carried out as part of a project concerned with empowerment in intensive care nursing. Viewed from a psychological perspective, empowerment is understood as referring to the ability to make goal directed decisions and to take action towards meeting the goals set out. Autonomy, then, is seen as part of empowerment. The data were collected with a structured questionnaire specially designed for this study. The random sample was drawn from 31 Finnish adult intensive care units. The respondents were registered nurses working in eight adult intensive care units. The data were analysed using statistical methods. Autonomy was defined as being composed of three bases: knowledge base (independence, right and responsibility in decision-making), action base (independence, right and responsibility in actions) and value base (independence, right and responsibility in values). The majority of the respondents felt they had more autonomy in decision-making and actions concerning patient care than in decision-making and actions concerning the intensive care unit as a whole. Autonomy usually increased with work experience. Work will be continued to study the autonomy of nurses in different health care organisations in Finland. PMID- 12590893 TI - Reviewing evidence. AB - This paper summarises how evidence based practice is defined, and what can be considered to constitute evidence. It describes the steps which should be undertaken in a systematic review of evidence and some of the issues involved in implementing the findings from such reviews in practice. This includes identifying the types of evidence which should inform decisions about practice and how these should be integrated with clinical expertise and client choice in order to achieve best practice. PMID- 12590894 TI - Achieving collaborative workplace learning in a university critical care course. AB - The purpose of this paper is to describe the development, implementation and evaluation of a new critical care curriculum based on the tenets of collaborative workplace learning. It also examines lecturers' and clinical educators' issues, and explores students' evaluations of the old curriculum compared with those of the new curriculum. Three data collection methods were used for this study. Comprehensive notes were maintained of the meetings conducted with lecturers, clinical educators and representative students during the development and implementation of the course. Three focus group interviews were conducted with students before the introduction of the new curriculum and three focus group interviews were conducted during first semester following implementation of the new curriculum. Quality-of-teaching surveys were also completed by two groups of critical care course students: one group before and one group following the introduction of the new curriculum. Major findings in this study included: developing a sense of ownership of the curriculum for clinical educators, clinical educators' difficulties with addressing their responsibilities, amalgamating theoretical learning with clinical practice, and tackling students' workload. This paper demonstrates the value of using the collaborative workplace learning approach in strategically addressing the challenges of developing and conducting a university critical care course. PMID- 12590895 TI - A new era in the treatment of coronary disease? PMID- 12590896 TI - Screening individuals and families with premature coronary heart disease: a clinical and public health challenge. PMID- 12590897 TI - The final common pathway. PMID- 12590898 TI - A meta-analytical approach for the treatment of in-stent restenosis. PMID- 12590899 TI - All that glitters is not gold. PMID- 12590900 TI - Mitochondria and the heart. PMID- 12590901 TI - Statin effects beyond lipid lowering--are they clinically relevant? PMID- 12590902 TI - Screening of family members of patients with premature coronary heart disease; results from the EUROASPIRE II family survey. AB - AIMS: To determine whether the Joint European Societies' recommendations that first degree blood relatives of patients with premature coronary heart disease (CHD) should be screened for coronary risk factors is being followed and, if so, how effectively these relatives are being managed. METHODS AND RESULTS: Using a postal questionnaire, 3322 relatives (siblings and children >/=18 years of age) of 1289 index patients in the EUROASPIRE II survey who had suffered from premature CHD (men under 55 years and women under 65 years) were asked whether screening for coronary risk factors had occurred and, if so, how they were being managed in terms of lifestyle advice and drug therapies. Overall, screening for coronary risk factors because of CHD in the family was only performed in 11.1% of siblings and 5.6% of children. However, prevalences of different cardiac risk factors were high both in relatives and offspring and a clear familial clustering could be documented. Less than 50% of siblings and 25% of children were given some general lifestyle advice regarding cardiac risk factors. Moreover, active interventions such as starting antihypertensive or lipid lowering drugs were rarely carried out, particularly in children of patients with premature CHD. CONCLUSIONS: European physicians rarely screen family members of patients with premature CHD for cardiac risk factors. General lifestyle style advice or active treatment for these risk factors are also rarely given. However, since these family members have a high prevalence and familial clustering of cardiac risk factors, they form an ideal target population for primary prevention of CHD in high-risk patients. PMID- 12590903 TI - Cardiogenic shock complicating acute myocardial infarction; prognostic impact of early and late shock development. AB - AIMS: Cardiogenic shock accounts for the majority of deaths following acute myocardial infarction. The majority of outcome data on this issue are, however, derived from single hospitals, referral centers or selected patients in randomized studies. The purpose of this study was to investigate incidence, outcome and prognostic significance of cardiogenic shock in 6676 consecutive patients with acute myocardial infarction. METHODS AND RESULTS: Demographic and clinical data including the presence of cardiogenic shock were prospectively collected in 6676 non-invasively managed patients with myocardial infarction consecutively admitted to 27 different hospitals during a 2-year period. Six-year mortality data were collected in 99.9% of the population. Cardiogenic shock developed in 444 patients (6.7%). In 59% of these patients cardiogenic shock developed within 48 h, 11% developed shock during days 3 and 4 and 30% later than 4 days after the infarction. Thirty-day and 6-year mortality was 62 and 88% among shock patients compared to 9 and 45% in non-shock patients. Patients with early shock development (days 1-2) had a significantly lower 30-day mortality (45%) than those with intermediate or late shock development (>80%) (P<0.05). In 30-day survivors, survival the following years was lower than in patients without cardiogenic shock but with post-infarction heart failure. CONCLUSIONS: In this nationwide prospectively collected registry, non-invasively managed consecutive myocardial infarct patients with cardiogenic shock had an extremely reduced life expectancy. Every attempt to improve treatment, prevention and identification of patients at risk of shock development should be strongly encouraged. PMID- 12590904 TI - Outcome after treatment of coronary in-stent restenosis; results from a systematic review using meta-analysis techniques. AB - AIMS: To evaluate the clinical outcome after treatment of coronary in-stent restenosis. METHODS AND RESULTS: For identification of the relevant literature a specific search strategy was conducted and explicit inclusion criteria were defined to avoid selection bias. Based on the selected literature, a systematic review using descriptive statistics and meta-analysis methods regarding the outcome after treatment of coronary in-stent restenosis was performed. The proportion of patients experiencing a major adverse cardiac event (MACE) as defined by death, myocardial infarction, and target lesion revascularization was the main outcome measure. A total of 1304 citations were identified. Among these, 28 studies (six different treatment modalities) including a total of 3012 patients met the inclusion criteria and were incorporated into this analysis. The estimated average probability of experiencing a major cardiac adverse event after treatment for in-stent restenosis with a follow-up period of 9+/-4 months was 30.0% (25.0-34.9%, 95% confidence interval) with strong evidence for heterogeneity between study specific results (P=0.0001). The clinical outcome was not significantly different between treatment modalities. After adjustment for confounding factors (i.e. lesion length), however, patients undergoing intracoronary radiation showed an estimated advantage of 16.9% (-37.7+/-4.0%, 95% confidence interval) in MACE free survival, as compared to balloon angioplasty. The post-interventional diameter stenosis was the only independent predictor for the long-term outcome after treatment of in-stent restenosis. CONCLUSIONS: Treatment of in-stent restenosis is associated with an overall 30% rate of major adverse cardiac events. Currently, repeat angioplasty is the treatment option of choice, especially when a sufficient acute procedural result can be achieved. Intracoronary radiation should be considered in cases with therapy refractory forms of diffuse in-stent restenosis. PMID- 12590905 TI - Survival after withdrawal of dofetilide in patients with congestive heart failure and a short baseline QTc interval; a follow-up on the Diamond-CHF QT substudy. AB - BACKGROUND: We have previously observed dofetilide to be associated with improved survival when the pre-treatment baseline QTc interval was below 429 ms. In this study we tested the natural extension of this observation-that the same group of patients should have a loss of survival benefit after withdrawal of dofetilide. METHODS: Patients with congestive heart failure (CHF) and reduced left ventricular function enrolled in the Diamond-CHF (Danish Investigations of Arrhythmia and Mortality on Dofetilide-CHF) study were eligible for our QT substudy provided they were in sinus rhythm and had a measurable QTc interval from a 12-lead standard ECG taken before randomization to placebo or dofetilide. An extended follow-up was performed from study closure, December 1996 until August 2001. RESULTS: Of the 418 patients entering the extended follow-up, 215 (51%) patients died during this 4.5 years of additional observation time. The baseline QTc interval made no prognostic difference to mortality in placebo treated patients. For dofetilide treated patients with a baseline QTc interval <429 ms, increased mortality was observed during the extended follow-up compared to placebo (risk ratio 1.5, 95% confidence interval 1.1-2.8)[corrected]. CONCLUSIONS: This follow-up study shows significant loss of survival benefit upon withdrawal of dofetilide in patients with CHF and a pre-treatment QTc interval below 429 ms. An independent randomized trial is warranted to validate these results. PMID- 12590906 TI - Cardiomyopathy in children with mitochondrial disease; clinical course and cardiological findings. AB - AIMS: To determine the frequency of cardiomyopathy in children with mitochondrial disease and describe their clinical course, prognosis and cardiological manifestations. METHODS AND RESULTS: Of 301 children with CNS and neuromuscular disease referred to our institution in 1984 to 1999, 101 had mitochondrial disease. Seventeen patients had cardiomyopathy, diagnosed by echo-Doppler investigations, all of the hypertrophic, non-obstructive type. The onset of symptomatic mitochondrial disease ranged from birth to 10 years of age. Eight children had cytochrome-c oxidase deficiency, while the remaining nine had various defects. Cardiomyopathy was diagnosed from birth to 27 years. Left ventricular posterior wall and septal thickness were both increased: z-scores +4.6+/-2.6 and +4.3+/-1.6 (mean+/-SD), respectively. The left ventricular diastolic diameter z-score, +1.3+/-3.4, and fractional shortening, 24+/-13%, displayed marked variations. Nine patients developed heart failure. Eleven patients with cardiomyopathy died, including all eight with cytochrome-c oxidase deficiency, and one patient underwent a heart transplantation. Mortality in children with mitochondrial disease was higher in those with cardiomyopathy (71%) than those without (26%) (P<0.001). CONCLUSIONS: In children with mitochondrial disease, cardiomyopathy was common (17%) and was associated with increased mortality. The prognosis for children with cytochrome-c oxidase deficiency and cardiomyopathy appeared to be particularly unfavorable. PMID- 12590907 TI - Syncope and coronary anomalies. PMID- 12590909 TI - Cerebral specialization and verbal-motor integration in adults with and without Down syndrome. AB - Persons with Down syndrome (DS) tend to exhibit an atypical left ear-right hemisphere advantage (LEA) for the perception of speech sounds. In the present study, a recent adaptation of the dichotic listening procedure was employed to examine interhemispheric integration during the performance of a lateralized verbal-motor task. Although adults with DS (n = 13) demonstrated a right ear-left hemisphere advantage in the dichotic-motor task similar to their peers with (n = 14) and without undifferentiated developmental disabilities (n = 14), they showed an LEA in a free recall dichotic listening task. Based on a comparison of the laterality indices obtained from both dichotic listening procedures, it appears that the manifestation of lateral ear advantages in persons DS may dependent on the response requirements of the task. PMID- 12590910 TI - Dissociations among functional categories in Korean agrammatism. AB - This study investigated the hypothesis that the syntactic trees formed by individuals with agrammatic aphasia cannot be constructed any higher than an impaired node as suggested by the tree-pruning hypothesis (Friedman, 1994; Friedmann & Grodzinsky, 1997) and hypothesis. It also examined their following implication that the members of a certain functional category are subject to the same degree of impairment. Two experiments were conducted to investigate a Korean agrammatic patient's use and understanding of three functional categories--Mood, Tense, and Complementizer. The results showed a dissociation among functional categories that preserves the higher node while leaving the lower node impaired both in production and comprehension. Another dissociation was found among members of the same category depending on their linear position in the clause. These results contrast with the predictions of the tree-pruning hypothesis, suggesting that the nearer to the end of the clause a functional element is located, the better it is preserved in Korean agrammatism. PMID- 12590911 TI - Brain imaging of tongue-twister sentence comprehension: twisting the tongue and the brain. AB - This study used fMRI to investigate the neural basis of the tongue-twister effect in a sentence comprehension task. Participants silently read sentences equated for the syntactic structure and the lexical frequency of the constituent words, but differing in the proportion of words that shared similar initial phonemes. The manipulation affected not only the reading times and comprehension performance, but also the amount of activation seen in a number of language related cortical areas. The effect was not restricted to cortical areas known to be involved in articulatory speech programming or rehearsal processes (the inferior frontal gyrus and anterior insula), but also extended to areas associated with other aspects of language processing (inferior parietal cortex) associated with phonological processing and storage. PMID- 12590912 TI - Hyperlexia profiles. AB - Two hyperlexic individuals were tested on reading outcome measures and on basic skills shown to be critical in reading acquisition. Results suggested distinct pathways to superior word recognition skills that can be described under a contemporary model of skilled reading, with differential acquisition of basic skills; superior word recognition skill was associated in both cases with specialized development in one basic area, that of orthographic processing. Results supported an asset rather than a deficit analysis of hyperlexia: Rather than assimilating to reading pathology (dyslexia) subtypes, hyperlexia was seen to be accompanied by assets that are absent in dyslexic profiles. PMID- 12590913 TI - On-line syntactic processing in aphasia: studies with auditory moving window presentation. AB - Twenty-eight aphasic patients with left hemisphere strokes and matched control subjects were tested on an auditory moving windows task in which successive phrases of a sentence were presented in response to subjects' self-paced button presses and subjects made timed judgments regarding the plausibility of each sentence. Pairs of sentences were presented that differed in syntactic complexity. Patients made more errors and/or took longer in making the plausibility judgments than controls, and were more affected than controls by the syntactic complexity of a sentence in these judgments. Normal subjects showed effects of syntactic structure in self-paced listening. On-line syntactic effects differed in patients as a function of their comprehension level. High-performing patients showed the same effects as normal control subjects; low performing patients did not show the same effects of syntactic structure. On-line syntactic effects also differed in patients as a function of their clinical diagnosis. Broca's aphasic patients' on-line performances suggested that they were not processing complex syntactic structures on-line, while fluent aphasics' performances suggested that their comprehension impairment occurred after on-line processing was accomplished. The results indicate that many aphasic patients retain their ability to process syntactic structure on-line, and that different groups of patients with syntactic comprehension disorders show different patterns of on-line syntactic processing. PMID- 12590914 TI - The effect of spectral manipulations on the identification of affective and linguistic prosody. AB - We investigated the effect of various spectral manipulations on the identification of sentential prosody. Two main categories of prosody--affective (happy, angry, sad) and linguistic (statement, question, continuation)--were studied. Thirty-six subjects were presented with stimuli that were recorded by a female native speaker of American English. The stimuli were digitally manipulated to create synthesized, band-pass filtered (F0-range and F2/F3-range) and re entrant (pitch only version of stimulus is convolved with a steady-state signal) conditions. Results of a forced-choice discrimination paradigm showed that, in general, performance is remarkably robust despite spectral manipulation, even when there is relatively little spectral information. However, performance was significantly degraded in the low band-pass and re-entrant conditions. These observations are discussed in light of the relevance of the fundamental frequency as well as syllabification for the analysis of prosodic information. PMID- 12590915 TI - Sex differences in semantic language processing: a functional MRI study. AB - Predictions based on two models of sex differences in cerebral organization of language were compared by examining fMRI patterns of 10 females and 9 males during a semantic processing task. Both groups displayed activation of left inferior frontal gyrus (IFG), left superior temporal gyrus (STG), and cingulate. Females, but not males, showed bilateral IFG and STG activation. Further analyses revealed females had less diffuse left activation and greater right posterior temporal and insula region activation than males. Results support both an interhemispheric and an intrahemispheric model of sex differences in language, suggesting that the models may not be mutually exclusive. PMID- 12590916 TI - On the production of irrelevant arguments in agrammatism. AB - We carried out two experiments to see whether there is a possibility that agrammatic aphasics have difficulty in selecting semantic relations of verbs in production. We used picture-description tasks which required subjects to describe pictures with a given verb where the number of objects or people is larger than that of the argument nouns of the verb. The results showed that all the agrammatic subjects produced "irrelevant arguments," i.e., noun phrases with a case particle attached to them which cannot be arguments of provided verbs nor adjuncts and cannot occur with those verbs. The judgment tests requiring the subjects to judge whether sentences with an irrelevant argument were correct or not showed that the subjects' judgment of the grammaticality of those sentences was not normal. The two experiments suggest a possibility that the agrammatic subjects have difficulty in identifying semantic roles of verbs at the level before mapping between semantic roles and grammatical roles. PMID- 12590917 TI - Verb and auxiliary movement in agrammatic Broca's aphasia. AB - Verb production in agrammatic Broca's aphasia has repeatedly been shown to be impaired by a number of investigators. Not only is the number of verbs produced often significantly reduced, but verb inflections and auxiliaries are often omitted as well (e.g., Bastiaanse, Jonkers, & Moltmaker-Osinga, 1996; Saffran, Berndt, & Schwartz, 1989; Thompson, Shapiro, Li, &Schendel, 1994, 1997). It has been suggested that these problems are, in part, caused by the fact that finite verbs need to be moved from their base-generated position to inflectional nodes in the syntactic tree (e.g., Bastiaanse & Van Zonneveld, 1998). Others have suggested that production deficits in agrammatism can be predicted based on the position that certain structures take in the syntactic tree (Friedmann & Grodzinsky, 1997; Hagiwara, 1995). If the former theory is correct, several predictions can be made. First of all, the discrepancy between production of finite verbs in the matrix and embedded clause that has been found for Dutch (Bastiaanse & Van Zonneveld, 1998) should not be observed in English, since the word order of the matrix and embedded clause are the same in the latter language. Second, if verb movement (including movement of auxiliaries) is problematic for speakers with agrammatic aphasia, then a hierarchy in the production of auxiliaries in yes/no questions, auxiliaries, and finite verbs in declarative sentences in English would be expected, since the former has been moved and the two latter are in base-generated position. In the present paper, these hypotheses were tested in a cross-linguistic study of Dutch and English. Results showed the position in the syntactic tree does not predict deficit patterns; rather the critical factor appears to relate to whether or not verb or auxiliary movement is required. PMID- 12590918 TI - Electromagnetic field generation by ATP-induced reverse electron transfer. AB - This paper describes a mechanism to explain low-level light emission in biology. A biological analog of the electrical circuitry, modeled on the parallel plate capacitor, traversed by a helical structure, required to generate electromagnetic radiation in the optical spectral range, is described. The charge carrier required for the emissions is determined to be an accelerating electron driven by an ATP-induced reverse electron transfer. The radial velocity component, the emission trajectory, of the moving charges traversing helical protein structures in a cyclotron-type mechanism is proposed to be imposed by the ferromagnetic field components of the iron in the iron-sulfur proteins. The redox systems NADH, riboflavin, and chlorophyll were examined with their long-wavelength absorption maxima determining the energetic parameters for the calculations. Potentials calculated from the axial velocity components for the riboflavin and NADH systems were found to equal the standard redox potentials of these systems as measured electrochemically and enzymatically. The mechanics for the three systems determined the magnetic moments, the angular momenta, and the orbital magnetic fluxes to be adiabatic invariant parameters. The De Broglie dual wave-particle equation, the fundamental equation of wave mechanics, and the key idea of quantum mechanics, establishes the wavelengths for accelerating electrons which, divided into a given radial velocity, gives its respective emission frequency. Electrons propelled through helical structures, traversed by biologically available electric and magnetic fields, make accessible to the internal environment the optical spectral frequency range that the solar spectrum provides to the external environment. PMID- 12590919 TI - Functional analysis of site-directed glycosylation mutants of the human equilibrative nucleoside transporter-2. AB - Protein glycosylation is important for nucleoside transport, and this has been demonstrated for the human equilibrative nucleoside transporter-1 (hENT1). It is not known whether glycosylation affects the functions of hENT2 or where hENT2 is glycosylated. We address these questions using N-glycosylation mutants (N48D, N57D, and N48/57D) and demonstrate that hENT2 is glycosylated at Asn(48) and Asn(57). Our results show that although the apparent affinities for [3H]uridine and [3H]cytidine of the mutants were indistinguishable from those of the wild type protein, N-glycosylation was required for efficient targeting of hENT2 to the plasma membrane. All mutants had a two- to threefold increase in IC(50) for dipyridamole. N57D and N48/57D, but not N48D, also had a twofold increase in IC(50) for NBMPR. We conclude that the relative insensitivity of hENT2 to inhibitors is primarily due to its primary structure and not to glycosylation. Glycosylation modulates hENT1 function, but is not required for hENT2. PMID- 12590920 TI - Determination of the membrane contact residues and solution structure of the helix F/G loop of prostaglandin I2 synthase. AB - From our topological arrangement model of prostaglandin I(2) synthase (PGIS) created by homology modeling and topology studies, we hypothesized that the helix F/G loop of PGIS contains a membrane contact region distinct from the N-terminal membrane anchor domain. To provide direct experimental data we have explored the relationship between the endoplasmic reticulum (ER) membrane and the PGIS F/G loop using a constrained synthetic peptide to mimic PGIS residues 208-230 cyclized on both ends through a disulfide bond with added Cys residues. The solution structure and the residues important for membrane contact of the constrained PGIS F/G loop peptide were investigated by high-resolution 1H two dimensional nuclear magnetic resonance (2D NMR) experiments and a spin label incorporation technique. Through the combination of 2D NMR experiments in the presence of dodecylphosphocholine (DPC) micelles used to mimic the membrane environment, complete 1H NMR assignments of the F/G loop segment have been obtained and the solution structure of the peptide has been determined. The PGIS F/G loop segment shows a defined helix turn helix conformation, which is similar to the three-dimensional crystallography structure of P450BM3 in the corresponding region. The orientation and the residues contacted with the membrane of the PGIS F/G loop were evaluated from the effect of incorporation of a spin-labeled 12-doxylstearate into the DPC micelles with the peptide. Three residues in the peptide corresponding to the PGIS residues L217 (L11), L222 (L16), and V224 (V18) have been demonstrated to contact the DPC micelles, which implies that the residues are involved in contact with the ER membrane in the native membrane-bound PGIS. These results provided the first experimental evidence to localize the membrane contact residues in the F/G loop region of microsomal P450 and are valuable to further define and understand the membrane topology of PGIS and those of other microsomal P450s in the native membrane environment. PMID- 12590921 TI - The active site of the molybdenum cofactor biosynthetic protein domain Cnx1G. AB - The final step of molybdenum cofactor biosynthesis in plants is catalyzed by the two-domain protein Cnx1. The G domain of Cnx1 (Cnx1G) binds molybdopterin with high affinity and transfers molybdenum to molybdopterin. Here, we describe the functional and structural characterization of structure-based Cnx1G mutants. For molybdopterin binding residues Thr542 and Ser573 were found to be important because different mutations of those residues resulted in 7- to 26-fold higher k(D) values for molybdopterin binding. Furthermore, we showed that the terminal phosphate of molybdopterin is directly involved in protein-pterin interactions as dephosphorylated molybdopterin binds with one magnitude of order lower affinity to the wild-type protein. Molybdopterin binding was not affected in mutants defective in Ser476, Asp486, or Asp515. However, molybdenum insertion was completely abolished, indicating their important role for catalysis. Based on these results we propose the binding of molybdopterin to a large depression in the structure of Cnx1G formed by beta5, alpha5, beta6, and alpha6, whereas the negatively charged depression formed by the loop between beta3 and alpha4, the N terminal end of alpha2, the 3(10) helix, and the region between beta6 and alpha6 is involved in catalysis. PMID- 12590922 TI - Relaxin regulates fibrillin 2, but not fibrillin 1, mRNA and protein expression by human dermal fibroblasts and murine fetal skin. AB - Relaxin modulates connective tissue remodeling by altering matrix molecule expression. We have found that relaxin specifically inhibits a microfibril component, fibrillin 2 (FBN2), without affecting fibrillin 1 (FBN1). Human dermal fibroblasts (HDFs) grown or stimulated to overexpress fibrillin expression were used to show that relaxin specifically down-regulated FBN2 mRNA and protein levels. Continuous exposure of HDFs to relaxin (30ng/ml) significantly (P<0.05) decreased fibrillin 2 protein (40%) while FBN1 protein expression was unchanged. Our in vitro studies were confirmed using relaxin null mice whereby the absence of relaxin was associated with increased FBN2 mRNA and protein in fetal skin from pregnant relaxin knockout mice. The regulation of FBN2 expression may be associated with functional changes in elastic tissues during development and growth. PMID- 12590923 TI - Heat effect on the structure and activity of the recombinant glutamate dehydrogenase from a hyperthermophilic archaeon Pyrococcus horikoshii. AB - Glutamate dehydrogenase from Pyrococcus horikoshii (Pho-GDH) was cloned and overexpressed in Escherichia coli. The cloned enzyme with His-tag was purified to homogeneity by affinity chromatography and shown to be a hexamer enzyme of 290+/ 8 kDa (subunit mass 48 kDa). Its optimal pH and temperature were 7.6 and 90 degrees C, respectively. The purified enzyme has outstanding thermostability (the half-life for thermal inactivation at 100 degrees C was 4 h). The enzyme shows strict specificity for 2-oxoglutarate and L-glutamate and requires NAD(P)H and NADP as cofactors but it does not reveal activity on NAD as cofactor. K(m) values of the recombinant enzyme are comparable for both substrates: 0.2 mM for L glutamate and 0.53 mM for 2-oxoglutarate. The enzyme was activated by heating at 80 degrees C for 1 h, which was accompanied by the formation of its active conformation. Circular dichroism and fluorescence spectra show that the active conformation is heat-inducible and time-dependent. PMID- 12590924 TI - Mice lacking inducible nitric oxide synthase show strong resistance to anti-Fas antibody-induced fulminant hepatitis. AB - Although nitric oxide (NO) plays important roles in pathogenesis of various liver diseases, the role of NO in the in vivo mechanism of Fas-mediated fulminant hepatitis is not known well. The effect of anti-Fas antibody (Jo2) on the survival, liver function, and histology was analyzed in wild-type (WT) and inducible NO synthase (iNOS)-deficient (iNOS(-/-)) mice. Upon intravenous injection of a lethal dose of Jo2, WT mice died on fulminant hepatitis within 12h. Under identical conditions, however, iNOS(-/-) mice showed strong resistance to Jo2 and survived without revealing liver injury. In conclusion, these observations suggest that regulation of NO metabolism may have therapeutic potential in the treatment of patients with fulminant hepatitis. PMID- 12590925 TI - Chain scission of hyaluronan by peroxynitrite. AB - The reaction of peroxynitrite with the biopolymer hyaluronan has been studied using stopped-flow techniques combined with detection of molecular weight changes using the combination of gel permeation chromatography and multiangle laser light scattering. From the effect of peroxynitrite on the yield of hyaluronan chain breaks, it was concluded that the chain breaks were caused by hydroxyl radicals which escape a cage containing the *OH NO*(2) radical pair. The yield of free hydroxyl radicals was determined as 5+/-1% (as a proportion of the total peroxynitrite concentration). At high peroxynitrite concentrations, it was observed that the yield of chain breaks leveled out, an effect largely attributable to the scavenging of hydroxyl radicals by nitrite ions present in the peroxynitrite preparation. These experiments also provided some support for a previous proposal that the adduct formed between ONOOH and ONOO(-) might itself produce hydroxyl radicals. The rate of this reaction would have to be of the order of 0.05 s(-1) to produce hydroxyl radical yields that would account quantitatively for chain break yields at high peroxynitrite concentrations. By carrying out experiments at higher hyaluronan concentrations, it was also concluded that an additional yield of chain breaks was produced by the bimolecular reaction of the polymer with ONOOH at a rate constant of about 10 dm(3)mol(-1)s(-1). At 5.3 x 10(-3)mol dm(-3) hyaluronan, this amounted to 3.5% chain breaks (per peroxynitrite concentration). These conclusions support the proposal that the yield of hydroxyl radicals arising from the isomerization of ONOOH to nitrate ions is relatively low. PMID- 12590926 TI - Differential effects of mutations in human endothelial nitric oxide synthase at residues Tyr-357 and Arg-365 on L-arginine hydroxylation and GN-hydroxy-L arginine oxidation. AB - Biosynthesis of nitric oxide (NO) is catalyzed by NO synthase (NOS) through a two step oxidation of L-arginine (Arg) with formation of an intermediate, GN-hydroxy L-Arg (NHA). In this study we have employed mutagenesis to investigate how residues Y357 and R365 which interact primarily with the substrate Arg and (6R) 5,6,7,8-tetrahydro-L-biopterin (H(4)B) modulate these two steps of the NOS reaction. Mutant Y357F preserved most wild-type heme characteristics and NADPH oxidation ability. However, mutation of this residue markedly increased the dissociation constants for both Arg and NHA by 20-fold and decreased the NO synthesis from Arg by 85% compared to that of wild type. Mutation of Y357 had less effect on the rate of NO generated from NHA. Mutant R365L purified in the presence of Arg had a normal heme environment and retained 9 and 55% of the wild type NO formation rate from Arg and NHA, respectively. When Arg was removed from buffer, R365L instantly became a low-spin state (Soret peak at 418 nm) with the resultant loss of H(4)B and instability of the heme-CO complex. The low-spin R365L exhibited an NADPH oxidation rate higher than that of wild type. Its Arg driven NO formation was decreased to near the limit of detection, whereas the rate of NHA-driven NO synthesis was one third that of wild type. This NHA-driven NO formation completely relied on H(4)B and was not sensitive to superoxide dismutase or catalase but was inhibited by imidazole. The wild-type eNOS required 14 microM NHA and 0.39 microM H(4)B to reach the half-maximal NHA-driven NO formation rate (EC(50)), while R365L needed 59 microM NHA and 0.73 microM H(4)B to achieve EC(50). The differential effect of mutation on Arg and NHA oxidation suggests that distinct heme-based active oxidants are responsible for each step of NO synthesis. PMID- 12590927 TI - A novel trypsin inhibitor from Peltophorum dubium seeds, with lectin-like properties, triggers rat lymphoma cell apoptosis. AB - A trypsin inhibitor (PDTI) was isolated from Peltophorum dubium seeds by affinity chromatography on a thyroglobulin-agarose or a trypsin-agarose column. In both cases, SDS-PAGE showed two bands of M(r) 20,000 and 22,000, which could not be resolved. Their amino-terminal sequences were identical and similar to that of Kunitz-type soybean trypsin inhibitor (SBTI). Mass spectrometry analysis of tryptic digests of both bands showed 16 coincident peaks, suggesting that they are closely related proteins. The K(i)s for trypsin and chymotrypsin inhibitory activity of PDTI were 1.6 x 10(-7) and 1.3 x 10(-5)M, respectively. Lectin-like activity of PDTI and SBTI, detected by hemagglutination of rabbit erythrocytes, was inhibited by sialic acid-containing compounds. PDTI and SBTI caused apoptosis of Nb2 rat lymphoma cells, demonstrated by decrease of viability, DNA hypodiploidy, DNA fragmentation, and caspase-3-like activity. They had no effect on normal mouse splenocytes or lymphocytes, whereas they caused apoptosis of concanavalin A-stimulated mouse lymphocytes. PMID- 12590928 TI - Rice HMGB1 protein recognizes DNA structures and bends DNA efficiently. AB - We analyzed the DNA-binding and DNA-bending properties of recombinant HMGB1 proteins based on a rice HMGB1 cDNA. Electrophoretic mobility shift assay demonstrated that rice HMGB1 can bind synthetic four-way junction (4H) DNA and DNA minicircles efficiently but the binding to 4H can be completed out by HMGA and histone H1. Conformational changes were detected by circular dichroism analysis with 4H DNA bound to various concentrations of HMGB1 or its truncated forms. T4 ligase-mediated circularization assays with short DNA fragments of 123 bp showed that the protein is capable of increasing DNA flexibility. The 123-bp DNA formed closed circular monomers efficiently in its presence, similar to that in an earlier study on maize HMG. Additionally, our results show for the first time that the basic N-terminal domain enhances the affinity of the plant HMGB1 protein for 4H DNA, while the acidic C-terminal domain has the converse effects. PMID- 12590929 TI - Chemical denaturation of a homodimeric lysine-49 phospholipase A2: a stable dimer interface and a native monomeric intermediate. AB - Bothropstoxin I (4BthTx-I) is a homodimeric lysine-49 (Lys49) phospholipase A(2) isolated from Bothrops jararacussu venom, which damages liposome membranes via a Ca(2+)-independent mechanism. The stability of the BthTx-I homodimer was evaluated by equilibrium chemical denaturation with guanidinium hydrochloride monitored by changes in the intrinsic tryptophan fluorescence anisotropy, far-UV circular dichroism, dynamic light scattering, and 1-anilinonaphthalene-8 sulfonate binding. Unfolding of the BthTx-I dimer proceeds via a monomeric intermediate with native-like structure, with Gibbs free energy (DeltaG(0)) values of 10.0 and 7.2 kcal mol(-1) for the native dimer-to-native monomer and native-to-denatured monomer transitions, respectively. The experimentally determined DeltaG(0) value for the dimer-to-native monomer transition is higher than the value expected for an interaction dominated by hydrophobic forces, and suggests that an unusually high propensity of hydrogen-bonded side chains found at the BthTx-I homodimer interface make a significant contribution to dimer stability. PMID- 12590930 TI - Effect of thyroid state on rate and sites of H2O2 production in rat skeletal muscle mitochondria. AB - The purpose of this study was to investigate the effects of thyroid state on rates and sites of H(2)O(2) production in rat muscle mitochondria. With Complex I and Complex II-linked substrates, hypothyroidism decreased and hyperthyroidism increased the rates of O(2) consumption during State 4 and State 3 respiration and the rates of H(2)O(2) release during State 4 respiration. During State 3, the rates of H(2)O(2) release were not affected by thyroid state. However, the mitochondrial capacity to remove H(2)O(2) increased in the transition from hypothyroid to hyperthyroid state, thus suggesting that an increase in H(2)O(2) production rate also occurred in such a transition during State 3 respiration. The observation that mitochondrial coenzyme Q levels and cytochrome oxidase activities are higher in the hyperthyroid and lower in the hypothyroid groups suggests that the modifications of H(2)O(2) production are due to a modulation by thyroid hormone of the mitochondrial content of autoxidizable electron carriers. This idea is supported by measurements of H(2)O(2) release in the presence of respiratory inhibitors. In fact, such measurements indicate that the thyroid state-linked changes in H(2)O(2) production occur at both generator sites of the respiratory chain. PMID- 12590931 TI - Mechanism of strong resistance of Helicobacter pylori respiration to nitric oxide. AB - The aim of the present work is to elucidate the mechanism by which the respiration of Helicobacter pylori but not of Escherichia coli shows a strong resistance to nitric oxide (NO). Nitric oxide strongly but reversibly inhibited the oxygen consumption by sonicated membranes from H. pylori and Triton X-100 treated cells. Although the sensitivity of the H. pylori respiration to cyanide was low, it also increased after the treatment with Triton X-100. Kinetic analyses revealed that NO was rapidly degraded by E. coli and the Triton X-100 treated H. pylori, but not by the intact H. pylori. Thus, the low sensitivity to NO might reflect the low affinity of the cytochrome c oxidase for this radical within the membrane/lipid bilayers of H. pylori. Such properties of the oxidase in H. pylori membranes may, at least in part, underlie the mechanism by which this bacterium thrives in NO-enriched gastric juice. PMID- 12590932 TI - Ascorbic acid blunts oxidant stress due to menadione in endothelial cells. AB - Endothelial cells are exposed to potentially damaging reactive oxygen species generated both within the cells and in the bloodstream and underlying vessel wall. In this work, we studied the ability of ascorbic acid to protect cultured human-derived endothelial cells (EA.hy926) from oxidant stress generated by the redox cycling agent menadione. Menadione caused intracellular oxidation of dihydrofluorescein, which required the presence of D-glucose in the incubation medium, and was inhibited by intracellular ascorbate and desferrioxamine. At concentrations of 100 microM and higher, menadione depleted the cells of both GSH and ascorbate, and ascorbate loading partially prevented the decrease in GSH due to menadione. Menadione increased L-arginine uptake by the cells, but inhibited endothelial nitric oxide synthase, an effect that was prevented by acute loading with ascorbate. Ascorbate blunts menadione-induced oxidant stress in EA.hy926 cells, which may help to preserve nitric oxide synthase activity under conditions of excessive oxidant stress. PMID- 12590933 TI - Autoxidation of extracellular hydroquinones is a causative event for the cytotoxicity of menadione and DMNQ in A549-S cells. AB - Cytotoxicity of 1,4-naphthoquinones has been attributed to intracellular reactive oxygen species (ROS) generation through one-electron-reductase-mediated redox cycling and to arylation of cellular nucleophiles. Here, however, we report that in a subclone of lung epithelial A549 cells (A549-S previously called A549-G4S (Watanabe, et al., Am. J. Physiol. 283 (2002) L726-736), the mechanism of ROS generation by menadione and by 2,3-dimethoxy-1,4-naphthoquinone (DMNQ), and therefore that of cytotoxicity, differs from the paradigm. Ninety percent of H(2)O(2) generation by both the quinones can be prevented by dicumarol, an inhibitor of NAD(P)H quinone oxidoreductase (NQO1), at the submicromolar level, regardless of the quinone concentrations. Exogenous SOD also inhibits H(2)O(2) production at low but not high concentrations of the quinones, especially DMNQ. Thus, at low quinone concentrations, superoxide-driven hydroquinone autoxidation accounts for more than half of H(2)O(2) generation by both quinones, whereas at high quinone concentrations, especially for DMNQ, comproportionation-driven hydroquinone autoxidation becomes the predominant mechanism. Hydroquinone autoxidation appears to occur predominantly in the extracellular environment than in the cytosol as extracellular catalase can dramatically attenuate quinone induced cytotoxicity throughout the range of quinone concentrations, whereas complete inactivation of endogenous catalase or complete depletion of intracellular glutathione has only a marginal effect on their cytotoxicity. Finally, we show evidence that ROS production is a consequence of the compensatory defensive role of NQO1 against quinone arylation. PMID- 12590934 TI - Chronic exposure to 2,5-hexanedione impairs the glutamate-nitric oxide-cyclic GMP pathway in cerebellar neurons in culture and in rat brain in vivo. AB - 2,5-Hexanedione is a neurotoxic metabolite of hexane. The mechanisms of its neurotoxicity remain unclear. We assessed whether chronic exposure to 2,5 hexanedione affects the glutamate-nitric oxide-cGMP pathway in primary cultures of cerebellar neurons and/or in the cerebellum of rats. Chronic exposure of cultured cerebellar neurons to 2,5-hexanedione (200 microM) reduced by approximately 50% NMDA-induced formation of cGMP. Activation of soluble guanylate cyclase by nitric oxide was reduced by 46%. This treatment reduced the content of neuronal nitric oxide synthase and soluble guanylate cyclase in neurons by 23 and 20%, respectively. In the cerebellum of rats chronically exposed to 2,5 hexanedione (in the drinking water) NMDA-induced formation of cGMP was reduced by 55% as determined by in vivo brain microdialysis. Activation of soluble guanylate cyclase by nitric oxide was reduced by 65%. The content of neuronal nitric oxide synthase and of soluble guanylate cyclase was reduced by 25 and 21%, respectively, in the cerebellum of these rats. The effects are the same in both systems, indicating that cultured neurons are a good model to study the mechanisms of neurotoxicity of 2,5-hexanedione. These results indicate that chronic exposure to 2,5-hexanedione affects the glutamate-nitric oxide-cGMP pathway at different steps both in cultured neurons and in cerebellum of the animal in vivo. The alteration of this pathway may contribute to the neurotoxic effects of 2,5-hexanedione. PMID- 12590935 TI - Serotonin transport is modulated differently by tetanus toxin and growth factors. AB - It has been previously shown that 5-HT uptake inhibition produced by tetanus toxin (TeTx) corresponds to a non-competitive inhibition, and it is preceded by phosphorylation of the tyrosine-kinase receptor trkA, phospholipase C activation and translocation of protein kinase C isoforms [FEBS Lett. 481 (2000) 177; FEBS Lett. 486 (2000) 136]. In the present work, it is shown that agonists of tyrosine kinase receptors (NGF, EGF, basic FGF) enhance Na(+)-dependent, 5 hydroxytryptamine (serotonin, 5-HT) uptake in the synaptosomal-enriched P(2) fraction from rat-brain, suggesting a divergence in the intracellular signal pathways triggered by TeTx and by agonists of TyrK receptors. Co-applications of TeTx and agonists of TyrK receptors result in a mutual and partial reversion of their effects on 5-HT transport. In spite of their differences on transport, TeTx, TPA and NGF produce an increase in serotonin transporter phosphorylation in Ser separately, which is abolished by the PKC-inhibitor bisindolylmaleimide-1. Co application of sodium vanadate, a tyrosine-phosphatase inhibitor, partially abolishes the effect produced by TeTx, whereas genistein, a tyrosine-kinase inhibitor, does not exert any variation of TeTx inhibition. Analyses by immunoblotting of the activation of specific PKC isoforms activation, determined as translocation to the membrane compartment, reveals differences in the pattern produced by NGF and TeTx. PKC gamma, delta, and epsilon isoforms are equally activated by both compounds, whereas the beta isoform is activated in a sustained manner only by TeTx, and the alpha isoform is only down-regulated by NGF. The aim of the present work was to explore whether NGF have the same effect on 5-HT transport than TeTx, since both compounds share the ability of activate part of the same transduction pathways. In spite of this, growth factors and TeTx show an opposite effect on 5-HT transport, even though SERT phosphorylation is enhanced in both cases. The differential effect on alpha- and beta-PKC isoenzymes found between NGF and TeTx action could explain this apparent discrepancy. PMID- 12590936 TI - Long-term exposition of cells to beta-amyloid results in decreased intracellular calcium concentration. AB - The ubiquitously present beta-amyloid peptide plays an important role in the pathogenesis of Alzheimer's disease. Its neurotoxicity has been blamed on its mal activity to increase calcium-levels. In the present study, we demonstrate that treatment of fibroblasts with beta-amyloid has, indeed, resulted in a transient rise in the calcium-concentration. Chronic exposition of cultures to the peptide, however, caused a fall in the calcium-level. Apparently, beta-amyloid has biphasic effects: acutely, it increases the calcium-concentration of cells; in contrast, on the long-run, beta-amyloid peptide acts as a calcium-antagonist. Therefore, the idea that beta-amyloid peptide leads to neural degeneration solely by increasing cells' calcium concentration must be replaced with a more complex view of its dual function in intracellular ionic homeostasis. PMID- 12590937 TI - Post-mortem degradation of brain glutamate decarboxylase. AB - The post-mortem stability of the GABA synthesizing enzyme glutamate decarboxylase (GAD) was studied by using SDS-PAGE and quantitative immunoblotting to measure the rates of degradation of GAD in the cerebral cortex, hippocampus, and cerebellum of rats and mice as a function of time after death. The intact 65- and 67-kDa isoforms of GAD (GAD(65) and GAD(67)) disappeared gradually over a 24-h period. In both rats and mice, the degraded GAD appeared as a band with an apparent molecular mass of 55-57 kDa; no significant amounts of smaller forms were observed. The 55-57 kDa band reacted with antiserum W887, which recognizes a shared epitope at the carboxyl-terminal end of both GADs, indicating that GAD was cleaved near the amino-terminal end of the molecule. GAD(67) was cleaved at a site between the amino-terminus and the epitope for antiserum W883 (located within residues 79-93 of GAD(67)), as antiserum W883 stained a 56-kDa band on the blots. The appearance of degraded GAD paralleled the loss of total GAD (GAD(65)+GAD(67)), and after 24h the 55-57 kDa band accounted for 97, 88, and 59% of the intact GAD lost from rat cerebellum, cerebral cortex and hippocampus. On a percentage basis, GAD(67) was degraded more rapidly than was GAD(65) in all brain regions studied. The loss of GAD activity was greater in rat than mouse brain, even though the percent loss of intact GAD protein was similar. PMID- 12590938 TI - Protein phosphatase type 2C dephosphorylates BAD. AB - Reversible phosphorylation modulates a cells' susceptibility to apoptosis. The phosphorylation status of BAD, a member of the Bcl-2 protein family, is an important checkpoint governing life-or-death decisions: Phosphorylation of serine residues 112, 136 and 155 on BAD prevents apoptosis. Here we report that BAD is a substrate for PP2C. Ser(155) is involved in heterodimerization with Bcl-X(L). We could demonstrate that PP1, PP2A and PP2C act on this site in vitro. However, only PP2C gives priority to P-Ser(155) compared to P-Ser(112) and P-Ser(136) on BAD. The results indicate that PP2C is an additional factor triggering the pro apoptotic function of BAD. PMID- 12590939 TI - Phosphinic, phosphonic and seleninic acid bioisosteres of isonipecotic acid as novel and selective GABA(C) receptor antagonists. AB - A number of amino acids bioisosterically derived from the specific GABA(A) agonist, isonipecotic acid, were electrophysiologically characterized as antagonists at GABA(C) rho(1) receptors expressed in Xenopus oocytes. The phosphinic acid analogue of isonipecotic acid, piperidin-4-ylphosphinic acid (2), was comparable with the standard GABA(C) antagonist, (1,2,5,6-tetrahydropyridin-4 yl)methylphosphinic acid (TPMPA), in terms of potency and GABA(C) versus GABA(A) receptor selectivity. Whereas the phosphonic acid analogue, piperidin-4 ylphosphonic acid (4), was at least an order of magnitude weaker than piperidin-4 ylphosphinic acid as a GABA(C) antagonist, the seleninic acid analogue, piperidin 4-ylseleninic acid (SEPI, 6), was the most potent and selective GABA(C) antagonist within the group of isonipecotic acid derived amino acids studied. PMID- 12590940 TI - Cytosine arabinofuranoside-induced activation of astrocytes increases the susceptibility of neurons to glutamate due to the release of soluble factors. AB - Activation of astrocytes occurs during many forms of CNS injury, but its importance for neuronal survival is poorly understood. When hippocampal cultures of neurons and astrocytes were treated from day 2-4 in vitro (DIV 2-4) with 1 microM cytosine arabinofuranoside (AraC), we observed a stellation of astrocytes, an increase in glial fibrillary acidic protein (GFAP) level as well as a higher susceptibility of the neurons to glutamate compared with cultures treated from DIV 2-4 with vehicle. To find out whether factors released into the culture medium were responsible for the observed differences in glutamate neurotoxicity, conditioned medium of AraC-treated cultures (MCMAraC) was added to vehicle treated cultures and conditioned medium of vehicle-treated cultures (MCMvh) was added to AraC-treated cultures 2 h before and up to 18 h after the exposure to 1mM glutamate for 1 h. MCMAraC increased glutamate neurotoxicity in vehicle treated cultures and MCMvh reduced glutamate neurotoxicity in AraC-treated cultures. Heat-inactivation of MCMvh increased, whereas heat-inactivation of MCMAraC did not affect glutamate toxicity suggesting that heat-inactivation changed the proportion of factors in MCMvh inhibiting and exacerbating the excitotoxic injury. Similar findings were obtained using conditioned medium of pure astrocyte cultures of DIV 12 treated from DIV 2-4 with vehicle or 1 microM AraC suggesting that heat-sensitive factors in MCMvh were mainly derived from astrocytes. Treatment of hippocampal cultures with 1mM dibutyryl-cAMP for 3 days induced an activation of the astrocytes similar to AraC and increased neuronal susceptibility to glutamate. Our findings provide evidence that activation of astrocytes impairs their ability to protect neurons after excitotoxic injury due to changes in the release of soluble and heat-sensitive factors. PMID- 12590941 TI - Phenylarsine oxide inhibits alpha-latrotoxin-stimulated [3H]GABA release from rat brain synaptosomes. AB - Phosphatidylinositol 4,5-biphosphate has been implicated in a variety of membrane trafficking processes, including exocytosis of neurotransmitters. However, there are contradictory findings concerned ability of phenylarsine oxide (PAO), an inhibitor of phosphatidylinositol 4-kinase, to affect exocytotic release of different types of neurotransmitters. We bent our efforts to a detailed analysis of action of PAO on Ca(2+)-dependent and Ca(2+)-independent [3H]GABA release produced by exposure of rat brain synaptosomes to different concentrations of alpha-latrotoxin. We also compared PAO action on alpha-latrotoxin- and 4 aminopyridine (4-AP)-evoked [3H]GABA release. The experiments have shown that release of [3H]GABA evoked by the depolarization with 4-AP was decreased by 80% as a result of action of 3 microM PAO and the complete inhibition of release was observed with 10 microM PAO. When alpha-latrotoxin as a stimulant was applied, release of [3H]GABA was increased as toxin concentration used was elevated from 0.5 to 3.0 nM, however, concomitantly, the response of the toxin-induced [3H]GABA release to PAO became attenuated: 10 microM PAO led to almost complete inhibition of the effect of 0.5 nM alpha-latrotoxin and only partly decreased (by 40%) the response to 3.0 nM alpha-latrotoxin. To test whether the efficacy of PAO depended on the toxin-induced outflow of cytosolic [3H]GABA, synaptosomes with depleted cytosolic [3H]GABA pool were also exploited. Depletion was performed by means of heteroexchange of cytosolic [3H]GABA with nipecotic acid. The experiments have shown that treatment of loaded synaptosomes with nipecotic acid resulted in some increase of [3H]GABA release evoked by 0.5 nM alpha-latrotoxin, but in the two fold decrease of the response to 3.0 nM alpha-latrotoxin. PAO essentially inhibited [3H]GABA release from depleted synaptosomes irrespective of alpha latrotoxin concentration used. Therefore, the amount of [3H]GABA released from cytosolic pool determined, in considerable degree, the insensitivity of alpha latrotoxin action to PAO. Thus, our data show that subnanomolar concentrations of alpha-latrotoxin may be used for stimulation of exocytotic release of [3H]GABA. Exposure of synaptosomes with nanomolar toxin concentrations leads not only to stimulation of exocytosis, but also to leakage of [3H]GABA from cytosolic pool. PAO potently inhibits exocytotic release of [3H]GABA and its inhibitory effectiveness is diminished as far as the outflow of [3H]GABA is elevated. PMID- 12590942 TI - Leptin-induced transactivation of NPY gene promoter mediated by JAK1, JAK2 and STAT3 in the neural cell lines. AB - Neuropeptide Y (NPY) plays an important role in the central and sympathetic regulation of food intake and blood pressure. Although the NPY gene expression is regulated by a number of agents such as leptin, the mechanism responsible for leptin-induced regulation of the transcription of the NPY gene remains to be explored. In this study, the NPY gene promoter was transactivated by leptin in N18TG2, NG108-15 and PC12 cells which expressed the functional leptin receptor. The long isoform of leptin receptor (OB-Rb) could induce the transactivation, but the C-terminal truncated form (OB-Ra) could not. When dominant negative type of STAT3, JAK1 or JAK2 and was co-expressed, the leptin-induced transactivation was suppressed almost completely. The leptin-response element which confers NPY gene transactivation by leptin was determined in the 221-bp region of rat NPY gene promoter (-553/-335), where two STAT3-binding site-like elements (TCCAGTA) exist. These results indicated that activation of JAK1, JAK2 and STAT3 is necessary for leptin-induced transactivation of NPY gene through the leptin-response element in these neural cells. PMID- 12590943 TI - Residual substantia nigra neuromelanin in Parkinson's disease is cross-linked to alpha-synuclein. AB - The pigmentation of substantia nigra pars compacta dopaminergic neurons is due to the presence of neuromelanin, an irregular macromolecular pigment belonging to the family of melanins. Depletion of neuromelanin in Parkinson's disease is typically indicated by loss of brown color in this area. Unlike that from controls, the pigment extracted from substantia nigra of parkinsonian patients seems to be mainly composed by highly cross-linked, protease-resistant proteic material and the neuromelanin macromolecule appears to be a minor presence. In the present paper we describe the isolation by SDS-PAGE of this proteic component after cleavage of the melanin backbone under solubilizing conditions. A single band is observed, which has been identified as alpha-synuclein by western blotting. As expected, the same process performed on a control specimen did not show occurrence of any major proteic component. Nevertheless, extraction from a 91 years old control with Lewy bodies displayed minor alpha-synuclein immunoreactive aggregates, whereas inclusion of free alpha-synuclein was not observed at all. Results reported here support the view that alpha-synuclein accumulates within substantia nigra neurons and is entrapped in pigment granules during neuromelanin biosynthesis, i.e. before the melanin depletion characteristic of Parkinson's disease starts. PMID- 12590944 TI - Treatment and prophylaxis of venous thromboembolism during pregnancy. AB - The treatment and prevention of deep vein thrombosis (DVT) and pulmonary embolism (PE) in pregnant patients is challenging for several reasons. Coumarins can cause embryopathy and other adverse effects in the fetus. Although unfractionated heparin and low-molecular-weight heparins, the cornerstones of initial therapy, are safe for the fetus, they can have significant maternal side effects, including osteoporosis and thrombocytopenia. Because they must be given parenterally, long-term administration is inconvenient. Further, although low molecular-weight heparins probably cause less maternal osteoporosis and thrombocytopenia than unfractionated heparin, the appropriate dosing regimens for prevention and treatment of thrombosis during pregnancy have not been established. In addition, there is a paucity of reliable information on the incidence of venous thromboembolism and the risk of recurrent thrombosis during pregnancy. This paper briefly reviews the areas of controversy and provides recommendations for the treatment and prophylaxis of acute deep vein thrombosis and pulmonary embolism in pregnant patients. PMID- 12590946 TI - Resistance to aspirin in vitro at rest and during exercise in patients with angiographically proven coronary artery disease. AB - BACKGROUND: Acetylsalicylic acid, or aspirin, is widely used in secondary prevention of coronary artery diseases, but the inhibition of platelet aggregation is not uniform in all individuals. OBJECTIVE: To investigate the prevalence of aspirin resistance at rest and during exercise in coronary artery disease patients. MATERIALS AND METHODS: Fifty patients with stable coronary artery disease were prospectively studied. All patients received aspirin (75-300 mg/day for >1 month) and no other antiplatelet therapy. Aspirin resistance was studied, at rest and immediately after a stress test, using the standardized platelet function analyzer (PFA-100(R), Dade-Behring). Aspirin resistance was defined as a normal collagen/epinephrine closure time (<186 s). RESULTS: Ten patients (20%) were aspirin-resistant at rest. Out of the 40 patients who were aspirin-sensitive at rest, 9 (22%) were aspirin-resistant immediately after the exercise stress test. There were no differences in aspirin sensitivity regarding gender, age, diabetes, hypertension, dyslipidemia, platelet count, medical treatment or number of the coronary arteries involved. CONCLUSIONS: Aspirin resistance is detected, at rest, in 20% of our patients with stable coronary artery disease. Aspirin treatment does not seem to protect against exercise induced platelet activation in 22% of such patients, despite aspirin sensitivity at rest. PMID- 12590945 TI - D-dimer, other markers of haemostasis activation and soluble adhesion molecules in patients with different clinical probabilities of deep vein thrombosis. AB - Two automated turbidimetric D-dimer assays (BC D-dimer Plus, Dade Behring, Marburg, Germany and Auto-Dimer, Biopool, Umea, Sweden) were compared to two enzyme-linked immunosorbent assays (ELISAs) (Enzygnost D-dimer micro, Dade Behring and Asserachrome D-dimer, Diagnostica Stago, Asnieres, France) and two rapid D-dimer assays (SimpliRed, Agen Biomedical, Brisbane, Australia and Minutex, Biopool) in out-patients with suspected deep vein thrombosis (DVT). In addition, the performance of prothrombin fragment 1+2 (F1+2), thrombin antithrombin complex (TAT), and soluble adhesion molecules VCAM-1 and P-selectin for DVT diagnosis was assessed. One hundred and thirty-five consecutive out patients with suspected DVT of the lower limb were included, and in 52 (39%), DVT was confirmed by compression ultrasound. All D-dimer assays investigated reliably excluded DVT in those patients without DVT irrespective of their pre-test clinical probability of DVT. One D-dimer ELISA (Dade Behring) gave the highest area under the receiver operating characteristic (ROC) curve compared to other assays, and therefore, this was the most accurate assay in differentiating patients with from patients without DVT. The diagnostic performance of one automated turbidimetric assay (Auto Dimer, Biopool) was similar to ELISA and its convenience close to rapid latex agglutination assays. Most patients with a high pre-test clinical probability of DVT had positive D-dimer regardless of the presence or absence of DVT, which decreased the specificity of the tests and made D-dimer determination less useful for this group of patients. Because the diagnostic accuracy [sensitivity, specificity, negative (NPV) and positive predictive value (PPV)] of F1+2, TAT, VCAM-1 and P-selectin was inferior to D dimer assay, these assays could not substitute or supplement D-dimer testing in diagnosis of DVT. Levels of VCAM-1 and P-selectin were increased in patients with DVT and should therefore be investigated further to clarify their role in DVT. PMID- 12590947 TI - Ultralarge von Willebrand factor multimers and normal ADAMTS13 activity in the umbilical cord blood. AB - INTRODUCTION: Recent studies demonstrate that deficiency of ADAMTS13, the metalloprotease that cleaves von Willebrand factor (VWF) in a shear-dependent manner, causes thrombotic thrombocytopenic purpura (TTP). Previously, ultralarge multimers of VWF were detected in the fetuses, the umbilical cords and the newborns. However, the significance of this finding is unknown. MATERIALS AND METHODS: The activity of ADAMTS13 and the multimer pattern of VWF in the cord blood, as well as the VWF antigenic and ristocetin cofactor levels, were analyzed using previously published methods. The presence of ultralarge multimers was determined by densitometric analysis. RESULTS: On the average, the level of VWF antigen and ristocetin cofactor activity are slightly increased in the newborns (mean+/-standard deviation, 1.66+/-0.76 and 1.45+/-0.64 U/ml, respectively). Ultralarge VWF multimers are detected in 11 of 17 umbilical cord plasma samples. However, the ADAMTS13 is normal (0.99+/-0.15 U/ml) in all but one sample in which it is mildly decreased (0.71 U/ml). CONCLUSIONS: Since ADAMTS13 activity is normal, we speculate that the presence of ultralarge multimers in the umbilical cord blood may be due to the low levels of shear stress in the umbilical circulation. These results may have important implications for understanding the manifestation of ADAMTS13 deficiency during the neonatal period. PMID- 12590948 TI - No association between the MTHFR A1298C and transcobalamin C776G genetic polymorphisms and hyperhomocysteinemia in thrombotic disease. PMID- 12590949 TI - Theonezolide A, a novel marine macrolide, induces drastic shape change in rabbit platelets by reorganization of microtubules. AB - Theonezolide A, a marine macrolide, and thrombin caused a shape change followed by an aggregation in the rabbit platelets. Theonezolide A-induced platelet shape change, estimated by a decrease in light transmission, appeared to a greater extent than thrombin-induced one. Morphological studies using an electron microscope showed that theonezolide A changed platelet shape with various numbers of long pseudopods, loosing their discoid shape. Theonezolide A-induced shape change was inhibited by a microtubule-stabilizing agent, taxol, but not by an actin-depolymerizing agent, cytochalasin B. In contrast, thrombin-induced shape change was inhibited by cytochalasin B but not by taxol. Confocal fluorescence microscopy showed that circumferential microtubule bundle disappeared in the platelets treated with theonezolide A. Theonezolide A had no direct effect on polymerization of microtubules isolated from bovine brain, indicating that it indirectly causes microtubule reorganization. These results suggest that theonezolide A induces drastic shape change through reorganization of microtubules in rabbit platelets. Thus, theonezolide A is a useful drug to examine microtubule reorganization in the cells. PMID- 12590950 TI - Markers of platelet activation and platelet-leukocyte interaction in patients with myeloproliferative syndromes. AB - INTRODUCTION: Changes in platelet count and function contribute to thrombo hemorrhagic episodes in chronic myeloproliferative syndromes (MPS). We used flow cytometry to study platelet-leukocyte conjugates and markers of platelet activation in patients with MPS. METHODS: Whole blood from patients with chronic myelogenous leukemia (CML), polycythemia vera (PV), chronic myelofibrosis (MF), and essential thrombocythemia (ET) and from healthy volunteers was prepared for flow cytometry. Platelet microparticles and platelet microaggregates were identified with anti-CD42b and forward scatter, activated platelets with anti CD62p. Anti-CD42b, anti-CD14, and anti-CD45 were used to study platelet-leukocyte conjugates. RESULTS: The percentage of CD62p-positive platelets was elevated in all myeloproliferate syndrome subtypes. The median percentage of platelet microparticles was 5.2% in controls and significantly higher in PV (12.0%), MF (11.0%), and ET (11.0%, all p<0.05). There was an increased percentage of platelet-neutrophil conjugates in patients with PV (8.3%) and ET (10.4%) compared to normal controls (6.8%, all p<0.05). Platelet-monocyte conjugates were 8.0% in controls and elevated in PV (15.4%) and ET (15.0%, all p<0.05). Patients with a history of venous or arterial thrombotic events had slightly less platelet leukocyte conjugates and slightly more microparticles than patients without thrombosis; however, this difference was not statistically significant. CONCLUSIONS: These findings suggest that platelet-leukocyte conjugate formation occurs in myeloproliferative syndromes and indicates platelet activation. Also, platelet microparticles are elevated and might provide a catalytic surface for thrombin generation. This could explain the clinical observation that patients with myeloproliferative syndromes have an increased risk to experience arterial or venous thrombotic events. PMID- 12590951 TI - The effect of submaximal exercise on platelet aggregation during late follicular and midluteal phases in women. AB - INTRODUCTION: The key role of platelets in the pathogenesis of atherosclerosis prompted considerable interest on the effect of physical exercise on platelets. Due to probable menstrual cycle variations, only a limited number of investigations have studied the effect of exercise on platelets in women. The study was undertaken to determine the effect of acute submaximal exercise on platelet aggregation and thromboxane A(2) (TxA(2)) formation in females during their late follicular and midluteal phases. MATERIALS AND METHODS: Twelve healthy, sedentary, female volunteers performed 15 min of cycling exercise at a workload that increased their heart rate to 75% of maximal in two phases of the menstrual cycle. The maximal rate of ADP and collagen-induced platelet aggregation was evaluated on citrated whole blood using the impedance technique. Thrombin-induced thromboxane A(2) formation was evaluated by the measurement of thromboxane B(2) (TxB(2)) level by enzyme-linked immunoassay. RESULTS AND CONCLUSION: No significant difference was found between maximal rates of platelet aggregation measured in the different phases of menstrual cycle. Collagen-induced platelet aggregation and platelet count increased significantly after the exercise in both late follicular and midluteal phases (p<0.05). ADP-induced platelet aggregation did not change due to the exercise during the two phases of menstrual cycle. The thromboxane B(2) level measured in the midluteal phase was significantly higher than that measured in late follicular phase at rest. It was significantly increased after the exercise in late follicular phase while no significant difference was found between pre-exercise and postexercise levels in the midluteal phase. The differences in thromboxane A(2) formation were pointed out in the changes in platelet reactivity status. The inhibitory systems for platelets need further investigations. Our findings support the idea that menstrual variations do not have pronounced and acute effects on both platelet aggregation and response of platelets to acute exercise. PMID- 12590953 TI - Aprotinin counteracts heparin-induced inhibition of platelet contractile force. AB - BACKGROUND: Aprotinin interferes with heparin binding to platelets and decreases blood loss during cardiopulmonary bypass (CPB). Heparin abolishes platelet force during CPB, and the extent of platelet force recovery after protamine administration appears to correlate with blood loss. This study assessed the effect of aprotinin on heparin suppression of platelet force. METHODS: Platelet force was measured using the Hemodyne Hemostasis Analyzer. Clots were formed from platelet-rich plasma (PRP) by the addition of batroxobin and 10 mM CaCl(2). Clotting conditions included pH 7.4, ionic strength 0.15 M, fibrinogen level 1 mg/ml and 75,000 platelets/microl. RESULTS: After 1200 s of clotting, force was reduced from 7110+/-1190 to 450+/-450 dyn by 0.2 U/ml of heparin. Platelet force in aprotinin [20 microg/ml (140 KIU/ml)] containing PRP was not suppressed by heparin addition (7480+/-2410 dyn). Aprotinin [40 microg/ml (280 KIU/ml)] addition to previously heparinized plasma counteracted heparin force suppression. Aprotinin (40 microg/ml) increased platelet force from 5630 to 11,138+/-562 in PRP devoid of heparin. Aprotinin did not affect thrombin activity, fibrin structure, platelet aggregation or secretion. CONCLUSIONS: Aprotinin counteracts heparin suppression of platelet force and enhances platelet force in the absence of heparin. Aprotinin-heparin-platelet interactions may help explain aprotinin's ability to reduce blood loss during CPB. PMID- 12590952 TI - Effect of the ingestion of Ginkgo biloba extract on platelet aggregation and urinary prostanoid excretion in healthy and Type 2 diabetic subjects. AB - Enhanced platelet function, particularly in response to collagen, is a common occurrence in diabetes that increases the risk of cardiovascular disease. Ginkgo biloba extract is ingested primarily to improve mental focus but it possesses a blood-thinning potential, which has not been well characterized. This study was designed to compare the effect of ingesting G. biloba extract on platelet aggregation in platelet-rich plasma (PRP) and prostanoid urinary excretion in healthy volunteers and subjects with Type 2 diabetes mellitus (T2DM). Before and after ingesting 120 mg of standardized G. biloba extract for 3 months, platelet aggregation was studied in PRP and urinary metabolites of thromboxane B(2) (TXB(2)) and prostacyclin (PGI(2)) were measured. In healthy volunteers (age, 42+/-11 years; BMI, 28.4+/-4.8 kg/m(2); n=28), the ingestion of G. biloba extract significantly increased fasting insulin and C-peptide (10+/-4 vs. 12+/-6 microU/ml, p<0.007 and 1.3+/-0.8 vs. 2.1+/-1.1 ng/ml, p<0.001, respectively) and significantly reduced collagen but not PAF-mediated platelet aggregation, converting 21 of 28 subjects with [COL+/EPI+] platelets to the [COL-/EPI+] phenotype. This was accompanied by a reduction of 11-dehydro-TXB(2) from 12.4+/ 6.1 to 10.3+/-6.1 ng/mg Cr (p<0.04) and PGI(2) metabolites (2,3-dinor-6-keto PGF(1alpha) and 6-keto-PGF(1alpha)) from 2.2+/-0.8 to 1.8+/-0.8 ng/mg Cr (p<0.05). In the T2DM subjects (age, 54+/-8; BMI, 36.6+/-7.9 kg/m(2); n=19), G. biloba ingestion did not affect pancreatic beta-cell function but significantly reduced platelet aggregation, converting 16 of 19 [COL+/EPI+] platelets to the [COL-/EPI+] phenotype. Unlike the healthy volunteers, this was not accompanied by a reduced urinary prostanoid excretion. G. biloba-induced reduction of both classes of prostanoid metabolites in healthy volunteers, but not in T2DM subjects, may suggest a nonselective inhibition of COX-1-mediated TXA(2) in platelets and COX-2-mediated PGI(2) production by the endothelial cells and perhaps platelet-enriched levels of arachidonic acid or COX-1 activity, or both, in T2DM subjects. PMID- 12590954 TI - Modulation of sphingosine 1-phosphate/EDG signaling by tumor necrosis factor alpha in vascular endothelial cells. PMID- 12590955 TI - Polymorphisms beta2-glycoprotein I: phospholipid binding and multimeric structure. AB - beta2-glycoprotein I is a phospholipid-binding protein of 326 amino acids and is found in plasma at a concentration of approximately 200 microg/ml. It has a sequence of positively charged amino acids located at the carboxy terminus that mediates anionic phospholipid binding. Two polymorphisms (306Cys-->Gly and 316Trp ->Ser) located at the phospholipid-binding site have been described. Homozygous state for either mutation and a compound heterozygous state show no phospholipid binding. Interestingly, heterozygotes for either 306Cys-->Gly or 316Trp-->Ser mutation have normal cardiolipin binding suggesting that beta2-glycoprotein I may circulate as a multimeric structure where wild-type subunits compensate the defective binding of the mutant ones. We investigated the effect of these mutations on quaternary structure of beta2-glycoprotein I and phospholipid binding. As previously reported, under native conditions, beta2-glycoprotein I shows an apparent molecular weight of approximately 320 kDa and it can be dissociated into subunits of lower molecular weight by boiling in 6 M urea. We show that the multimeric structure is not affected by the presence of mutations in the phospholipid-binding domain. beta2-glycoprotein I induces aggregation of anionic phospholipid vesicles suggesting again a multivalent interaction where at least two binding sites are required to bridge adjacent vesicles. beta2 glycoprotein I-induced aggregation does not cause vesicle fusion or damage as demonstrated by fluorescence resonance energy transfer (FRET) or encapsulated calcein release. In conclusion, the normal cardiolipin binding in heterozygous state for mutations at phospholipid-binding domain may be due to the multimeric structure of beta2-glycoprotein I. PMID- 12590956 TI - Regulation of clot retraction by glycoprotein IIb/IIIa antagonists. AB - Binding of fibrinogen to platelet glycoprotein (GP) IIb/IIIa induces clot retraction. Significant differences among GP IIb/IIIa antagonists were previously noted to inhibit thromboelastography in whole blood specimens. The relationship between efficacy of these agents and inhibition of clot retraction is unclear. Here, we use a plasma-free clot retraction assay to evaluate potency of GP IIb/IIIa antagonists to inhibit clot retraction and modulate platelet signaling, and to address whether these effects are realized in the clinically relevant dose range. The potencies for inhibition of clot retraction and aggregation are similar for antagonists with high affinity for resting platelets and slow off rates, whereas lower affinity and fast off-rate antagonists are disproportionately less effective in blocking clot retraction. A positive correlation is observed between inhibition of clot retraction and inhibition of tyrosine dephosphorylation across a number of GP IIb/IIIa antagonist pharmacophores. For lower affinity and fast off-rate antagonists, the concentrations required for inhibition of clot retraction clearly exceed the clinical dose range. Site occupancy studies combined with clot retraction experiments addressed whether high affinity and slow off-rate compounds can alter clot retraction during the dosing interval. Binding studies using [3H] Roxifiban, a high affinity GP IIb/IIIa antagonist, indicate that occupancy of >95% of GP IIb/IIIa sites is required to inhibit clot retraction. This level of occupancy is not routinely achieved in the clinic and is not tolerated, at least for chronic therapy. These results suggest that inhibition of clot retraction is not necessary for efficacy of GP IIb/IIIa antagonists. PMID- 12590957 TI - Swimming and hemostasis during rehabilitation in patients with coronary heart disease. PMID- 12590958 TI - ABO blood group distribution and severity of coronary artery disease among patients undergoing coronary artery bypass surgery in Northern Finland. PMID- 12590959 TI - Evolutionary immunobiology: new approaches, new paradigms. PMID- 12590960 TI - Workshop report: evolutionary immunobiology--new approaches, new paradigms. PMID- 12590961 TI - Comparative study of the strategies evolved by two parasitoids of the genus Asobara to avoid the immune response of the host, Drosophila melanogaster. AB - Asobara tabida and Asobara citri are two braconid endoparasitoids of Drosophila melanogaster larvae. We studied and compared the strategies evolved by these two species to avoid the immune reaction of their host. A. tabida has no negative impact on host cellular defenses and its eggs avoid encapsulation by adhering to host tissues. At the opposite, we found that A. citri, whose eggs are devoid of adhesive properties, affects the host encapsulation abilities, hemolymph phenoloxidase activity and concentrations of circulating hemocytes. Some of these effects could directly rely on a severe disruption of the hematopoietic organ anterior lobes observed in parasitized larvae. This is the first report of the immune suppressive abilities of a parasitoid from the Asobara genus. Results are presented and discussed with respect to the strategies of virulence evolved by other parasitoids to counteract the D. melanogaster immune system. PMID- 12590962 TI - Expression of penaeidin antimicrobial peptides in early larval stages of the shrimp Penaeus vannamei. AB - Penaeidins are a family of antimicrobial peptides constitutively produced and stored in the hemocytes of penaeid shrimps. We have determined the expression and the localization of penaeidins in the first early larval stages (Nauplius V, Zoea I, II, III and Mysis II) and in post-larvae (1 and 8) of the shrimp Penaeus vannamei. Using in situ hybridization and immunohistochemical analyses, we localized penaeidin transcripts and peptides in a few hemocytes of larvae from Mysis II stage. However, RT-PCR analyses showed that penaeidin mRNAs are already present in the early stage of Nauplius V. In addition, penaeidin expression could not be detected in other cells than hemocytes. Our observations highlight the potential involvement of penaeidins during larval development and the ontogeny of immune system through hemocytes during this period of life, where shrimps are particularly susceptible to infectious diseases. PMID- 12590963 TI - Amoebapores, archaic effector peptides of protozoan origin, are discharged into phagosomes and kill bacteria by permeabilizing their membranes. AB - Antimicrobial peptides are widespread in animal species and their function as defensive molecules may even have appeared before the evolution of metazoa. The amoeboid protozoon Entamoeba histolytica discharge membrane-permeabilizing polypeptides named amoebapores into the phagosome in which engulfed bacteria are situated as evidenced here by confocal laser microscopy and electron microscopy using specific antibodies. We demonstrate that the purified three isoforms of the amoebic polypeptides exhibit complementary antibacterial activities in vitro. The potency of amoebapores were compared with that of antimicrobial peptides of phylogenetically widespread species by monitoring in parallel their activities against representatives of gram-positive and gram-negative bacteria and liposomes in various assays, and differences in the mechanism of membrane permeabilization became apparent. Northern blot analysis revealed that expression of genes coding for amoebapores and amoebic lysozymes is not dramatically changed upon co-culture of amoebae with bacteria indicating that the antimicrobial arsenal is rather constitutively expressed than induced in these primitive phagocytes. PMID- 12590964 TI - A novel antimicrobial peptide from the sea hare Dolabella auricularia. AB - The sea hare Dolabella auricularia is a marine shell-less gastropod. Four cytotoxic glycoproteins named dolabellanin A, C, E and P were found in the animal previously. An antimicrobial factor was newly isolated from the sea hare's body wall including skin and mucus. This factor is a novel peptide which consists of 33 amino acid residues, and is called dolabellanin B2. Dolabellanin B2 was cytotoxically effective against some pathogenic microorganisms at a concentration of 2.5-100 microg/ml. PMID- 12590965 TI - CpG oligodeoxynucleotides activate grass carp (Ctenopharyngodon idellus) macrophages. AB - In mice and humans, B cells, antigen-presenting cells including monocytes, macrophages and dendritic cells and natural killer cells can be stimulated directly or indirectly by the bacterial DNA and oligodeoxynucleotides (ODN) containing the CpG motifs (CpG DNA). Using head kidney macrophages of grass carp (Ctenopharyngodon idellus) as an in vitro model, we investigated the effects of several CpG-ODNs on fish immunocytes. The CpG-ODNs included the optimal motifs: the ODN-1826 (GACGTT) and -2006 (GTCGTT) for the mice and humans cells, the ODN 1670 (AACGTT) used in Atlantic salmon, the ODN-D containing two repeats motif of those in 1670 and the ODN-R with an inverted CpG. The results showed that CpG has an immunomodulatory role in grass carp, and all the ODNs except the ODN-R could activate macrophages, increasing the levels of superoxide anion (O(2)(-)), hydrogen peroxide (H(2)O(2)), acid phosphatase and bactericidal activity. New evidence was provided that CpG-ODN could induce the cell-mediated immunity in fish. Interestingly, no significant differences among the ODNs tested could be found and the ODN-D was not more efficient than 1670. It suggests that the sequence which contains the unmethylated 'CG' dinucleotides could make contribute to this immunostimulatory effect. These findings indicate that CpG-ODNs could be useful tools for understanding the important anti-bacterial defense mechanism in fish. And it may have potential application in the minimizing the impact of fish diseases and enhancing the efficacy of antigen and DNA vaccines. PMID- 12590966 TI - Adaptive cell-mediated cytotoxicity against allogeneic targets by CD8-positive lymphocytes of rainbow trout (Oncorhynchus mykiss). AB - Rainbow trout surface-(s)IgM(-) leukocytes exhibited cell-mediated cytotoxicity (CMC) against allogeneic cells. This is described in concordance with a characterization of gene expression in the effector cells. Peripheral blood leukocytes (PBL) isolated from trout grafted with allogeneic tissue lysed allogeneic target cells (erythrocytes or cells of the RTG-2 cell line) in in vitro assays. The PBL were magnetically separated into different subpopulations using monoclonal antibodies (mabs) specific to thrombocytes, IgM, granulocytes and monocytes. Of the isolated subpopulations only the sIgM(-) lymphocytes were capable of lysing allogeneic targets. The separated PBL fractions were characterized by RT-PCR analysis using specific primers for the amplification of trout IgM heavy chain constant region (CH1), T cell receptor alpha chain (TCRalpha), CD8alpha and major histocompatibility complex (MHC) class I gene fragments. Most importantly, CD8alpha was expressed only by the sIgM(-) population. Combined with the requirement for sensitization to detect CMC, this strongly suggests T cell involvement in fish as in higher vertebrates. The involvement of CD8alpha-positive cytotoxic T cells in allograft rejection was supported by additional in vivo and in vitro observations. CD8alpha expression was barely detectable in the blood of unsensitized trout or trout that received xenografts, but was easily detected in the blood of allogeneically stimulated trout. Furthermore, CD8alpha expression in sIgM(-) lymphocytes from immunized trout was secondarily enhanced by addition of allogeneic targets in vitro. Collectively, these functional and genetic data suggest that fish possess specific cytotoxic cells with phenotype and gene expression pattern similar to those of cytotoxic T cells in higher vertebrates. PMID- 12590967 TI - An impairment of phagocytic function is linked to a shorter life span in two strains of prematurely aging mice. AB - In previous cross-sectional studies on Swiss mouse populations we have shown that at the same chronological age, animals that take longer to explore a simple T maze (slow mice) are hyper-emotional and show an impairment of the immune system than those which quickly explore the maze (fast mice). Therefore, we have proposed that the slow mice are a model of prematurely aging mice (PAM). In the present work we have carried out a longitudinal study of age-dependent changes in key functions of phagocytic cells (peritoneal macrophages) such as phagocytosis and superoxide anion production in both male and female Swiss (outbred strain) and BALB/c (inbred strain) PAM and non-prematurely aging mice (NPAM). Gender differences were found showing the females better phagocytic and digestion capacities with concomitant longer life span. The PAM showed an impaired phagocytosis capacity and intracellular superoxide anion production as well as an increase of its extracellular production as compared to NPAM, which could be related to the shortened life span of those animals in both sexes and strains. PMID- 12590968 TI - Molecular cloning of the beta subunit of complement component eight of rainbow trout. AB - Complement-mediated killing of pathogens through the lytic pathway is an important effector mechanism of the innate immune response. C8 is one of the components of the lytic pathway and is composed of an alpha, beta, and gamma subunit. In the present study we report the cloning and characterization of the primary structure of the C8beta subunit in the rainbow trout (Oncorhynchus mykiss). The deduced amino acid sequence of trout C8beta shows 72 and 47% identity with that of Japanese flounder and human, respectively. It also contains many of the same structural motifs as those found in mammalian lytic components. The C8beta gene appears to exists as a single copy in the trout genome and is expressed primarily in the liver. The protein encoded by the gene was identified by Western blotting using an anti-peptide antibody and was approximately 65kDa. PMID- 12590969 TI - The FREP gene family in the snail Biomphalaria glabrata: additional members, and evidence consistent with alternative splicing and FREP retrosequences. Fibrinogen related proteins. AB - Fibrinogen-related proteins (FREPs) found in hemolymph of the snail Biomphalara glabrata are hypothesized to be involved in non-self recognition. Among 150 cloned FREP cDNAs examined, we have identified three additional FREP members, FREPs 3.3, 12.1 and 13.1, bringing the total of FREP subfamilies to 13. The new FREPs each encode two immunoglobulin superfamily domains and a fibrinogen domain. Additionally, five truncated cDNAs with >99% nucleotide identity in coding regions to FREPs 3.2, 12.1 or 13.1 were identified. The truncated forms, the first reported for FREPs, lack a partial exon, one complete exon, or two complete exons plus the 3'UTR. Our preferred hypothesis is that all five truncated cDNAs observed arise from alternative splicing of full-length FREP genes. Genomic sequences lacking at least two introns and corresponding to the 3' ends of the cDNAs of FREP12.1 and its two truncated forms were also recovered. Although these could be the source of the truncated cDNAs, they are believed to be retrosequences. PMID- 12590970 TI - Manduca sexta lipopolysaccharide-specific immulectin-2 protects larvae from bacterial infection. AB - We previously reported the isolation of a lipopolysaccharide (LPS)-specific immulectin-2 from the tobacco hornworm, Manduca sexta [J. Biol. Chem. 275 (2000) 37373]. Immulectin-2 is a C-type lectin that is present at a constitutively low level in hemolymph of naive larvae, and its synthesis is induced after injection of Gram-negative bacteria or LPS. Immulectin-2 contains two carbohydrate recognition domains. It binds to LPS and stimulates prophenoloxidase activation in plasma. In this paper, we focus on properties of carbohydrate recognition domain-2 of immulectin-2 and the biological functions of immulectin-2 in immune responses. The carboxyl-terminal carbohydrate recognition domain (CRD2) of immulectin-2 was able to bind bacterial LPS. Binding of recombinant CRD2 to LPS stimulated activation of prophenoloxidase in plasma. Injection of antiserum against immulectin-2 into M. sexta larvae inhibited clearance of a Gram-negative bacterial pathogen, Serratia marcescens, and decreased survival of infection. These results suggest that immulectin-2 plays an important role in the immune system of M. sexta, and helps to protect the animal from Gram-negative bacterial infections. PMID- 12590971 TI - Nitric oxide production by carpet shell clam (Ruditapes decussatus) hemocytes. AB - We have demonstrated that carpet shell clam (Ruditapes decussatus) hemocytes produce nitric oxide (NO) in response to zymosan or bacterial lipopolysaccharide (LPS). This NO production was partially inhibited by the NO synthase inhibitor, N omega-nitro-L-arginine (L-NAME). The capability of clam hemocytes to produce NO in response to the bacterial pathogen Vibrio tapetis was also studied. Incubation with bacteria induced a significant NO production by clam hemocytes, even though exogenous NO only slightly decreased the growth of V. tapetis. The effect of exogenous NO on the capability of clam hemocytes to phagocytose labeled Escherichia coli was studied using two different NO donors S-nitroso-N-acetyl penicillamine (SNAP), and diethylenetriamine NO adduct (DETA/NO). Exogenous NO did not increase hemocyte phagocytosis, indicating that NO does not mediate phagocytosis in this species. These results are in accordance to those observed in other mollusk species, in which NO was independent of phagocytosis and constitutes an alternative method of killing invading pathogens. PMID- 12590972 TI - Identification of immunorelevant genes from greater wax moth (Galleria mellonella) by a subtractive hybridization approach. AB - In this study we have analyzed bacterial lipopolysaccharide (LPS) induced genes in hemocytes of the Lepidopteran species Galleria mellonella using subtractive hybridization, followed by suppressive PCR. We have found genes that show homologies to molecules, such as gloverin, peptidoglycan recognition proteins and transferrin known to be involved in immunomodulation after bacterial infection in other species. In addition, a few molecules previously not described in the innate immune reactions were detected, such as a RNA binding molecule and tyrosine hydroxylase. Furthermore, the full-length cDNA of a LPS-induced molecule with six toxin-2-like domains is described to be a promising candidate to further elucidate the relationship between toxin- and defensin-like domains in arthropod host defense. PMID- 12590973 TI - Characterization of rainbow trout and zebrafish eukaryotic initiation factor 2alpha and its response to endoplasmic reticulum stress and IPNV infection. AB - The cDNAs of rainbow trout and zebrafish eIF2alpha have been isolated and found to encode proteins of similar molecular weight and isoelectric point to the alpha subunit of the human translational initiation factor, eIF2. The rainbow trout (36.0kDa) and zebrafish (36.2kDa) eIF2alphas share 93 and 91% identity to the human protein, respectively, and are recognized by antibodies raised to the human form. In mammals, the phosphorylation of the alpha-subunit of eIF2 plays a key role in the regulation of protein synthesis in response to a range of cellular stresses. Regions corresponding to the human phosphorylation and kinase-docking sites are identical in the proteins of both fish species, as are residues that interact with the eIF2 recycling factor, eIF2B. Moreover, both recombinant rainbow trout and zebrafish eIF2alphas can be phosphorylated in vitro by the mammalian heme-sensitive eIF2alpha-kinase, HRI/HCR, as well as the interferon inducible, dsRNA sensitive kinase, PKR. Phosphorylation of rainbow trout and zebrafish eIF2alpha can also occur in vivo. RTG-2 and ZFL cells subjected to endoplasmic reticulum (ER) stress by treatment with the Ca(2+)-ionophore A23187 showed increased levels of eIF2alpha phosphorylation, suggesting similarity between the ER stress response in fish and other higher eukaryotes. Furthermore, RTG-2 cells responded to treatment with poly(I).poly(C) or to infection by infectious pancreatic necrosis virus, IPNV, by increasing eIF2alpha phosphorylation. These data imply that RTG-2 cells express the interferon-induced eIF2alpha-kinase, PKR and suggests that the interferon/eIF2alpha/PKR response to virus infection may be a conserved vertebrate characteristic. Overall these data are consistent with the premise that fish are able to regulate protein synthesis in response to cellular stresses through phosphorylation of eIF2alpha. PMID- 12590974 TI - Daily handling stress reduces resistance of carp to Trypanoplasma borreli: in vitro modulatory effects of cortisol on leukocyte function and apoptosis. AB - Carp subjected to daily handling stress were much more susceptible to Trypanoplasma borreli infection than control fish. In a search for the cellular mechanisms involved, it was observed that cortisol suppressed T. borreli-induced expression of interleukin-1beta, tumor necrosis factor-alpha, serum amyloid A and inducible nitric oxide synthase. An NF-kappaB-inhibitor could replicate cortisol induced apoptosis of activated peripheral blood leukocytes. In contrast, although this NF-kappaB-inhibitor induced apoptosis of neutrophilic granulocytes, cortisol prevented apoptosis of these cells, suggesting the latter process to be NF-kappaB independent. Carp leukocytes, upon induction of apoptosis, exhibit a number of sequential metabolic alterations. First, the mitochondrial transmembrane potential (DeltaPsi(m)) is disrupted and glutathione levels are depleted, followed by exposure of phosphatidylserine on the outer cell membrane. In vitro, cortisol could inhibit NO production induced by low concentrations of lipopolysaccharide (LPS), but remarkably, enhanced NO production induced by high concentrations of LPS. However, no differences in NO production were observed in stressed versus non-stressed infected carp. PMID- 12590975 TI - Characterization of beta(2)-microglobulin coding sequence from three non placental mammals: the duckbill platypus, the short-beaked echidna, and the grey short-tailed opossum. AB - To further characterize genes of immunological importance from non-placental mammals, cDNAs encoding beta(2)-microglobulin (beta(2)m) were isolated from two prototherians, the platypus and an echidna, and one metatherian, a grey short tailed opossum. In addition, a second allele of beta(2)m was identified in another metatherian species, the brushtail possum. Analysis of the deduced translations revealed conservation of key residues in these molecules over a long evolutionary history. The types of nucleotide substitutions present among the various taxa are also consistent with purifying selection at this conserved locus. An evolutionary tree of beta(2)m was constructed that supports the classic view of evolution with prototherians as the basal mammalian group. PMID- 12590976 TI - Characterization of monoclonal antibodies recognizing HLA-G or HLA-E: new tools to analyze the expression of nonclassical HLA class I molecules. AB - Nonclassical major histocompatibility complex (MHC) class I human leukocyte antigen E (HLA-E) and HLA-G molecules differ from classical ones by specific patterns of transcription, protein expression, and immunotolerant functions. The HLA-G molecule can be expressed as four membrane-bound (HLA-G1 to -G4) and three soluble (HLA-G5 to -G7) proteins upon alternative splicing of its primary transcript. In this study, we describe a new set of monoclonal antibodies (mAbs) called MEM-G/01, -G/04, -G/09, -G/13, MEM-E/02, and -E/06 recognizing HLA-G or HLA-E. The pattern of reactivity of these mAbs were analyzed on transfected cells by flow cytometry, Western blotting, and immunochemistry. MEM-G/09 and -G/13 mAbs react exclusively with native HLA-G1 molecules, as the 87G mAb. MEM-G/01 recognizes (similar to the 4H84 mAb) the denatured HLA-G heavy chain of all isoforms, whereas MEM-G/04 recognizes selectively denatured HLA-G1, -G2, and -G5 isoforms. MEM-E/02 and -E/06 mAbs bind the denatured and cell surface HLA-E molecules, respectively. These mAbs were then used to analyze the expression of HLA-G and HLA-E on freshly isolated cytotrophoblast cells, on the JEG-3 placental tumor cell line, and on cryopreserved and paraffin-embedded serial sections of trophoblast tissue. These new mAbs represent valuable tools to study the expression of HLA-G and HLA-E molecules in cells and tissues under normal and pathologic conditions. PMID- 12590977 TI - Interaction of HLA-DR with actin microfilaments. AB - Capping of HLA-DR on the surface of a human lymphoblastoid cell line (RAJI) and a transfectant human fibroblast cell line (M1DR1) was studied by confocal microscopy. Capping was induced at 22 degrees C after treating cells with an HLA DR specific monoclonal antibody, L243, followed by a secondary antibody conjugated with FITC. Cytoskeletal actin filaments (F-actin) accumulated under the caps were detected by rhodamine-phalloidin fluorescence. Two processes appear to take place: in the round lymphoblastoid cells, actin, initially distributed uniformly at the cell periphery, redistributes and becomes concentrated underneath HLA-DR patches or caps. In the non-round, substrate-attached fibroblasts, actin was organized in tightly packed filaments along the plasma membrane. It was observed that crosslinked HLA-DR receptors were associated with these filaments and were dragged toward the perinuclear area of the cells, where they coalesce to form a cap. The cytoskeleton-disrupting drugs that inhibit actin polymerisation were used to investigate the mechanism of capping of HLA-DR molecules. Sodium nitroprusside, a nitric oxide releasing agent, cytochalasin D both inhibited the percentage of capping in a dose-dependent manner. These data suggest that on antigen presenting cells, such as B cells and fibroblasts, actin microfilaments acts as a regulator of the movement and capping of HLA-DR receptors. PMID- 12590978 TI - Cytokine gene polymorphisms moderate illness severity in infants with respiratory syncytial virus infection. AB - Illness severity and frequency of complications in infants with respiratory syncytial virus (RSV) infection may be influenced by the local elaboration of cytokines. Cytokine gene polymorphisms moderate severity of illness in various inflammatory and infectious diseases. We performed cytokine genotyping on 77 infants hospitalized with confirmed RSV infection to determine whether specific cytokine gene polymorphisms are associated with illness severity or complications. DNA was extracted from buccal brushings and assayed for tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-6 and IL 10, and transforming growth factor (TGF)-beta1 genotypes using polymerase chain reaction-sequence-specific primer technology. Clinical outcomes consisted of severity scores of lower respiratory illness, blood oxygen saturation, lengths of oxygen supplementation, and intensive care unit (ICU) and hospital stays, and the presence or absence of pneumonia and otitis media. IFN-gamma genotype was related to severity of lower respiratory illness, duration of ICU stay, and frequency of otitis media. Additionally, IL-6 genotype was related to the length of oxygen (O(2)) supplementation and hospital stay, IL-10 genotype to the frequency of pneumonia, and TGF-beta1 genotype to O(2) saturations at presentation. There were no associations between TNF-alpha genotype and any of the outcome parameters. These results demonstrate that certain cytokine gene polymorphisms contribute to illness severity and complications during RSV infection in infants. If future prospective studies confirm these observations, cytokine genotyping may be a useful tool for identifying "at risk" infants who may benefit from the selective use of preventive or early intervention treatments for RSV. PMID- 12590979 TI - Intercellular adhesion molecule-1 K469E polymorphism: study of association with multiple sclerosis. AB - Intercellular adhesion molecule-1 (ICAM-1) is involved in the pathogenesis of multiple sclerosis (MS), whereas sequence variations in the ICAM-1 gene could potentially be responsible for the genetic susceptibility to MS. We studied an association of MS with the 13,848A>G (K469E) polymorphism of the ICAM-1 gene in Finnish and Spanish cases and controls and affected families. An increased risk for the AA (Lys(469)/Lys(469)) genotype was found in both populations. The effect observed was found to be strongest among the HLA-DQB1*0602-positive subjects, which implies genetic heterogeneity of MS. Meta-analysis of all published datasets supports increased risk of MS for the ICAM-1 Lys(469) homozygotes (relative risk = 1.3, p = 0.002). PMID- 12590980 TI - A collaborative European search for non-DQA1*05-DQB1*02 celiac disease loci on HLA-DR3 haplotypes: analysis of transmission from homozygous parents. AB - The HLA-DQA1*05 with DQB1*02 alleles are a major risk factor for celiac disease (CD). To search for additional human leukocyte antigen (HLA) risk factors, we looked on the DR3-DQ2 risk haplotype, selected because it carries both DQ risk alleles in cis and is the more represented among CD patients. In a European consortium, we identified 109 families with a parent homozygous for DQA1*05 DQB1*02. We typed ten microsatellites in the extended HLA complex, and applied the homozygous-parent transmission disequilibrium test (HPTDT) and extended-TDT to transmissions from homozygous parents. These methods eliminate confounding due to linkage disequilibrium between candidate disease loci and the known risk factor DQA1*05-DQB1*02, and are favorable when sufficient families are available. We did not find evidence of association with any single marker or allele, although weak evidence for additional risk was observed, represented by preferential transmission of six adjacent markers. We tested the largest ever reported HPTDT population in CD, providing unprecedented power. We did not find significant evidence of additional risk-modifying factors on the DR3 haplotype, independent of DQA1*05-DQB1*02, although a weak tendency was observed for the B8 DR3 haplotype. This effect should be tested in large populations with significant representations of both B8-DR3 and non-B8 DR3 haplotypes. PMID- 12590981 TI - Atopic asthma and TNF-308 alleles: linkage disequilibrium and association analyses. AB - The association of a tumor necrosis factor -308 allele (TNF2) to asthma has been reported in some studies but not in others. The aim of this study was to test this association in a population recruited on the basis of allergy to Parietaria. In the study population, asthma was positively associated to HLA-DRB1*03 (p = 0.01) and to the haplotype TNF2/DRB1*03 (p = 0.02). In the parent subgroup, the proportion of asthmatics was increased in patients with TNF2 (p = 0.01), but the primary association of asthma was to the haplotype TNF2/DRB1*1104 (p = 0.005). The study population was subdivided according to prick skin test (ST) positivity to Lolium, Parietaria, and D. pteronyssinus. Asthma was associated to HLA-DRB1*03 and to the haplotype TNF2/DRB1*03 (p = 0.0015 and 0.0001, respectively) in patients ST positive to Lolium, and to the haplotype TNF2/DRB1*1104 (p = 0.025) in patients ST positive to Parietaria. The transmission disequilibrium test detected excess transmission of HLA-DRB1*03 and of the haplotype TNF2/DRB1*03 (p = 0.03 and 0.04, respectively) to siblings with asthma and ST positivity to Lolium and of HLA-DRB1*1104 and of the haplotype TNF2/DRB1*1104 (p = 0.04 and 0.015, respectively) to siblings with asthma and ST positivity to Parietaria. Taken together, these observations indicate that the haplotypes TNF2/DRB1*03 and TNF2/*B1*1104 contain alleles controlling atopic asthma in patients with sensitization to Lolium and Parietaria, respectively. This suggests that the association of asthma to TNF2 reflects linkage disequilibrium with genes influencing specific immune response. PMID- 12590982 TI - Manganese superoxide dismutase and cytochrome P450 1A1 genes polymorphisms in rheumatoid arthritis in Taiwan. AB - To investigate the role of manganese superoxide dismutase (MnSOD) and cytochrome P450 1A1 (CYP1A1) gene polymorphisms in the pathogenesis of rheumatoid arthritis (RA) in Taiwan, MnSOD and CYP1A1 genes polymorphisms were determined by he polymerase chain reaction/restriction fragment length polymorphism method in 112 patients with RA and 96 controls. There were no significant differences in the genotype, allele, and phenotype frequencies of MnSOD Ala-9Val (C1183T) polymorphisms between patients with RA and controls. The polymorphism of MnSOD 5777T, threonine at the 58th amino acid, cannot be found in RA patients and controls in Taiwan. The allele and phenotype frequencies of CYP1A1 4887A and genotype frequency of CYP1A1 4887C/A were lower in RA patients than in controls, whereas the significant difference was lost after correction. MnSOD C1183T polymorphisms were not associated with the clinical manifestations of RA. However, RA patients with CYP1A1 4889G/G have significantly higher frequency of Sjogren's syndrome, especially in the presence of MnSOD 1183T/T. Patients with CYP1A1 4887C/A also have a trend to develop Sjogren's syndrome in the presence of MnSOD 1183T/T. The linkage disequilibrium between CYP1A1 4889G and CYP1A1 6235C can be found in this study. MnSOD gene polymorphisms are not related to susceptibility to RA in Taiwan, whereas individuals with CYP1A1 4887A tend to avoid the development of RA. Moreover, CYP1A1 4889G/G and 4887C/A may play a role in the development of Sjogren's syndrome, especially in the presence of MnSOD 1183T/T. These findings are preliminary. A further confirmation study is necessary. PMID- 12590983 TI - Genetic variation in the promoter region of the basic fibroblast growth factor gene. AB - Basic fibroblast growth factor (bFGF) is a member of the fibroblast growth factor family that possess broad mitogenic and cell survival activities and is involved in a variety of biological processes. We investigated possible genetic polymorphism in the promoter and 5' flanking region of the bFGF gene. Polymorphism was analysed by means of heteroduplex analysis, fragments with altered mobility were sequenced. Three novel substitutions (-553T/A, -834T/A and 921C/G) were identified in the promoter region. Allele frequencies in a sample of healthy Caucasian subjects (n=126) were determined by polymerase chain reactions followed by restriction analyses with specific endonucleases. The frequencies of the mutated alleles (-553A, -834A and -921G) were 0.04, 0.05 and 0.14, respectively. Newly identified variants in the bFGF gene promoter appear to be common polymorphisms in the Czech population. PMID- 12590984 TI - Distribution of MICA alleles and haplotypes associated with HLA in the Korean population. AB - The MICA (MHC class I chain-related gene A) is a polymorphic gene located 46 kb centromeric of the HLA-B gene, and is preferentially expressed in epithelial cells and intestinal mucosa. The MICA gene, similar to human leukocyte antigen (HLA) class I, displays a high degree of genetic polymorphism in exons 2, 3, 4, and 5, amounting to 54 alleles. In this study, we investigated the polymorphisms at exons coding for extracellular domains (exons 2, 3, and 4), and the GCT repeat polymorphism at the transmembrane (exon 5) of MICA in 199 unrelated healthy Koreans. Eight alleles were observed in the Korean population, with allele frequencies for MICA*010, MICA*00201, MICA*027, MICA*004, MICA*012, MICA*00801, MICA*00901, and MICA*00701 being 18.3%, 17.8%, 13.6%, 12.3%, 11.1%, 10.8%, 10.6%, and 3.3%, respectively. Strong linkage disequilibria were also observed between the MICA and HLA-B gene-MICA*00201-B58, MICA*004-B44, MICA*00701-B27, MICA*00801 B60, MICA*00901-B51, MICA*010-B62, MICA*012-B54, and MICA*027-B61. In the analysis of the haplotypes of HLA class I genes (HLA-A, B, and C) and the MICA, the most common haplotype was MICA*004-A33-B44-Cw*07, followed by MICA*00201-A2 B58-Cw*0302 and MICA*012-A2-B54-Cw*0102. The MICA null haplotype might be identified in the HLA-B48 homozygous individual. These results will provide an understanding of the role of MICA in transplantation, disease association, and population analyses in Koreans. PMID- 12590985 TI - Nomenclature for factors of the HLA system, updated November 2002. PMID- 12590986 TI - Nomenclature for factors of the HLA system, update December 2002. PMID- 12590987 TI - Tools, teamwork, and tenacity: an examination of family practice office system influences on preventive service delivery. AB - BACKGROUND: Most research examining primary care office characteristics and preventive service delivery (PSD) has evaluated preventive service aids and equipment, while generally overlooking the complex interactions among multiple office systems where multiple factors influence the overall practice. We test a theoretical model of practice influences on PSD that accounts for Tools (preventive service aids/equipment), Teamwork (office organization), and Tenacity (prevention delivery attitudes). METHODS: Office characteristics and 4454 patient visits were observed for 138 family physicians in northeast Ohio. Utilizing U.S. Preventive Services Task Force recommendations, age- and gender-specific PSD summary scores were computed for each patient and then averaged per physician. Multivariate analysis of variance tested office characteristic associations with PSD scores. RESULTS: Tools were common, but most were not significantly associated with PSD scores. The Teamwork indicators of clear staff role expectations and shared vision among physician and staff existed, respectively, for 80 and 73% of physicians. A high average reported practice focus on prevention existed, despite low staff involvement in PSD (22.2%). Compared with Tools, more Teamwork and Tenacity characteristics were associated with the PSD scores. CONCLUSION: Teamwork and Tenacity appear to be more important than Tools in delivering preventive services in primary care practices. PMID- 12590988 TI - Effect of provider status on preventive screening among Korean-American women in Alameda County, California. AB - BACKGROUND: Previous research suggests that having a doctor of the same ethnicity may be associated with lower rates of breast and cervical cancer screening in some Asian-American women. This study analyzes the effect of having a Korean, non Korean, or no regular doctor upon several measures of screening among Korean American women. METHODS: A random sample of 339 Korean-American women in Alameda County, California, were surveyed by telephone. Contingency tables and multivariable logistic regression were used to evaluate the association between provider status and six measures of recent screening, controlling for insurance and demographics. RESULTS: Having a non-Korean doctor was associated with an increased likelihood of having a Pap smear (odds ratio = 2.19, 95% confidence interval = 1.00, 4.80), mammogram (odds ratio = 7.63, 95% confidence interval = 2.35, 24.84), and clinical breast examination (odds ratio = 3.76, 95% confidence interval = 1.54, 9.20) in the past 2 years, compared to having a Korean doctor. This relationship is less apparent for nonfemale specific screening tests like cholesterol exams and routine checkups. CONCLUSIONS: Women who have a Korean doctor have less than optimal rates of breast and cervical cancer screening compared to women who have a non-Korean doctor. Having a Korean doctor may indicate less access to preventive health services, and programs to increase screening should target both Korean physicians and their female patients. PMID- 12590989 TI - Comparison of tailored interventions to increase mammography screening in nonadherent older women. AB - BACKGROUND: Recent increases in mammography use have led to a decrease in mortality from breast cancer. METHODS: Building on the Health Belief Model, the Transtheoretical Model, and past effectiveness of tailored interventions, we conducted a prospective randomized trial (n = 773) to test the efficacy on mammography adherence of tailored interventions delivered by five different methods, i.e., telephone counseling, in-person counseling, physician letter, and combinations of telephone with letter and in-person with letter. RESULTS: All five interventions increased mammography adherence significantly relative to usual care (odds ratios, 1.93 to 3.55) at 6 months post intervention. The combination of in-person with physician letter was significantly more effective than telephone alone or letter alone. Women thinking about getting a mammogram at baseline were more likely to be adherent by 6 months; even those in usual care achieved 48% adherence compared with 50-70% in the intervention groups. In contrast, women not thinking about getting a mammogram needed the interventions to increase their adherence from 13% to over 30%. CONCLUSIONS: All five interventions were effective at increasing mammography adherence. Women not thinking about getting a mammogram were most likely to benefit from these tailored interventions while other women might need less intensive interventions. PMID- 12590990 TI - National survey by specialty of U.S. physicians' HPV screening practices. AB - BACKGROUND: High-risk types of HPV are the primary cause of cervical cancer. OBJECTIVES: This article reports HPV screening and diagnosis from a survey evaluating community-based physicians' screening, testing, and clinical practices for sexually transmitted diseases. METHODS: Surveys mailed to physicians (n = 7,300) obtained information on patients they screen for HPV and cases of HPV diagnosed. RESULTS/CONCLUSIONS: Seventy percent (70%) of the physicians returned completed surveys. HPV screening was most frequently conducted in female patients by obstetrician/gynecologists and family practice physicians. PMID- 12590991 TI - Increased effectiveness of targeted skin cancer screening in the Veterans Affairs population of Northern California. AB - BACKGROUND: Skin cancer screening in populations at increased risk may be more useful than mass screening. We assessed the effectiveness of screening a targeted population in the Veterans Affairs Palo Alto Health Care System (VAPAHCS) for skin cancer/precancer detection and follow-up. METHODS: We studied the demographics, presumptive diagnoses, and outcome of 374 participants in free screening clinics conducted over a 3-year period in multiple northern California sites. The number of attendees with presumptive actinic keratosis (AK), basal cell carcinoma (BCC), squamous cell carcinoma (SCC), dysplastic nevus (DN), and melanoma was noted. RESULTS: Three hundred sixty-two males and 12 females were screened (mean age 63.4 years); 74% were Caucasian. Two hundred three individuals (54%) had a positive screen including 139 (52%) with presumptive AK, 41 (11%) with BCC, 9 (2%) with SCC, and 14 (4%) with DN versus potential melanoma. One hundred one (50%) of referred individuals were subsequently evaluated by VAPAHCS dermatologists. Biopsy was performed in 34/36 cases (94%), with a positive predictive value of 62% in patients with suspected BCC, 43% for SCC, 37.5% for DN and 12.5% for melanoma. CONCLUSIONS: Targeting a predominantly elderly Caucasian population with minimal to no prior dermatologic care yielded high rates of detection for precancers, skin cancer, and atypical nevi, and resulted in an increased percentage of pathologically confirmed nonmelanoma skin cancer, particularly BCC, compared to prior screening studies and population-based cancer registries. PMID- 12590992 TI - Smoking among adolescents in Muenster, Germany: increase in prevalence (1995 2000) and relation to tobacco advertising. AB - BACKGROUND: Understanding patterns and trends of smoking among youths is of major importance for the assessment of the burden of smoking in the society and efforts to decrease it. OBJECTIVE: The aims were to determine the prevalence and trends of smoking among adolescents in Muenster, Germany, and to assess its relation to youths' awareness of tobacco advertisement. DESIGN: Information on smoking habits was collected during two school-based surveys (1994/1995 and 1999/2000) of 12- to 15-year-old adolescents (3934 students in 1994/5 and 4028 students in 1999/2000) in Muenster, Germany. In addition, in 1994/1995 information about youths' awareness and appreciation of tobacco advertisement was collected. RESULTS: During the 5-year period, the prevalence of current smoking increased from 21.3 to 28.3%, and daily smoking increased from 10.0 to 14.2%. Among girls, daily smoking increased by 62% during this period. Almost all participants (94%) in 1994/1995 knew a tobacco brand, and appreciation of tobacco advertisement was strongly associated with the frequency of smoking. CONCLUSION: Smoking is increasing among adolescents in Germany especially among girls. These data are a cause of concern and call for efforts to reverse these trends, which should include a ban on tobacco advertisement in Germany. PMID- 12590993 TI - Social capital and the miniaturization of community among daily and intermittent smokers: a population-based study. AB - BACKGROUND: The impact of social participation, trust, and the miniaturization of community on daily and intermittent smoking was investigated. METHODS: The 2000 public health survey in Scania is a cross-sectional study. A total of 13,715 persons answered a postal questionnaire, which represents 59% of the random sample. A logistic regression model was used to investigate the association between the social capital variables and daily and intermittent smoking. The multivariate analysis was performed by using a logistic regression model to investigate the importance of possible confounders (age, country of origin, education, and snuff consumption) on the differences in daily and intermittent smoking between high versus low social participation, trust, and their four combination categories. The differences in the prevalences of the 13 social participation subitems between the high social capital and miniaturization of community categories were compared by t tests. RESULTS: Daily smoking is negatively associated with both social participation and trust, while intermittent smoking is positively associated with social participation and negatively associated with trust. This latter combination, named "the miniaturization of community," is an indirect measure of the ideologically and culturally increasingly narrow forms of social participation that excludes generalised trust to other people. Study circles, meetings of organisations, theatre/cinema, arts exhibition, and gathering of relatives are more prevalent in the high social capital category, while visit(s) to night club/entertainment is more prevalent in the miniaturization of community category. CONCLUSIONS: Low social capital is associated with daily smoking. "The miniaturization of community," i.e., high social participation and low trust, is significantly associated with intermittent smoking. The results have direct implications for smoking prevention strategies. PMID- 12590994 TI - An academic detailing intervention to disseminate physician-delivered smoking cessation counseling: smoking cessation outcomes of the Physicians Counseling Smokers Project. AB - BACKGROUND: Little is known about the effectiveness of interventions to disseminate smoking cessation interventions among a population of primary care physicians. This study's objective was to determine the effect of a community based academic detailing intervention on the quit rates of a population-based sample of smokers. METHODS: This community-based, quasi-experimental study involved representative samples of 259 primary care physicians and 4295 adult smokers. An academic detailing intervention was delivered to physicians in intervention areas over a period of 15 months. Analyses were performed on the data from the 2346 subjects who reported at least one physician visit over 24 months. Multivariate regression analyses were conducted to determine the impact of the intervention on self-reported smoking quit rates, reported by adjusted odds ratios. RESULTS: Among smokers reporting a physician visit during the study period, there was a borderline significant effect for those residing in intervention areas versus control areas (OR = 1.35; 95% CI.99-1.83; P = 0.057). Among a subgroup of 819 smokers who reported a visit with an enrolled physician, we observed a significant effect for those residing in intervention areas (OR = 1.80; 95% CI 1.16-2.75; P = 0.008). CONCLUSION: An academic detailing intervention to enhance physician delivered smoking cessation counseling is an effective strategy for disseminating smoking cessation interventions among community-based practices. PMID- 12590995 TI - How important are parents and partners for smoking cessation in adulthood? An event history analysis. AB - BACKGROUND: The aim of this study is to assess the effect of parental and partner's education and smoking behavior on an individual's chance of smoking cessation over the life course. METHODS: Self-reported life histories of smoking behavior, education, and relationships were recorded in face-to-face interviews with a random general-population sample of 850 respondents and their partners (if present). The data were collected in 2000. A discrete-time event history model is applied in the analyses of cessation over the life course. RESULTS: Parents' education and smoking behavior (during adolescence) and partners' education have no significant influence on cessation. Living with an ex-smoker or never-smoker increases the likelihood of quitting, compared to being single or living with a partner who smokes. Respondents whose partners were ex-smokers are almost five times more likely to quit smoking than single respondents. They are almost twice as likely to quit compared to those living with a never-smoker. CONCLUSIONS: The difference between having and not having a partner seems as important for cessation as the difference between having a partner who smokes, has never smoked, or has stopped smoking. An ex-smoking partner stimulates cessation more than a partner who has never smoked. Studies into cessation should take into account partners' smoking histories. PMID- 12590996 TI - Smoking cessation interventions in U.S. racial/ethnic minority populations: an assessment of the literature. AB - BACKGROUND: Smoking prevalence rates in some ethnic minority groups are elevated relative to the majority population. Thus, identifying cessation interventions that are effective for these groups is important. This article reviews published studies that examine effects of smoking cessation interventions relevant to racial ethnic minority populations. METHODS: A literature search of tobacco interventions, reporting smoking cessation outcomes (including quit rates) in U.S. minority populations, was conducted for the period 1985 to 2001. RESULTS: Thirty-six studies met preset criteria for inclusion. Twenty-three reported quit rates for African Americans, 4 for Asian/Pacific Islanders, 3 for Native Americans, and 10 for Hispanics. CONCLUSIONS: The disproportionate number of studies that focused on African American smokers compared with the other major racial/ethnic groups suggests the need for continued efforts to develop and evaluate the effectiveness of smoking cessation interventions for all ethnic minority populations. Abstinence rates varied considerably depending on study design and intervention strategy. Moreover, a relatively small percentage of studies that were randomized trials reported statistically significant findings, and most used intervention strategies that do not reflect the current state-of the-art. These results strongly suggest that more research is needed to identify successful smoking cessation interventions in these populations. PMID- 12590997 TI - Cardiovascular risk factor profiles and their social gradient from adolescence to age 74 in a Swiss region. AB - BACKGROUND: Few European studies have investigated how cardiovascular risk factors (CRF) in adults relate to those observed in younger generations. OBJECTIVE: To explore this issue in a Swiss region using two population health surveys of 3636 adolescents ages 9-19 years and 3299 adults ages 25-74 years. METHODS: Age patterns of continuous CRF were estimated by robust locally weighted regression and those of high-risk groups were calculated using adult criteria with appropriate adjustment for children. RESULTS: Gender differences in height, weight, blood pressure, and HDL cholesterol observed in adults were found to emerge in adolescents. Overweight, affecting 10-12% of adolescents, was increasing steeply in young adults (three times among males and twice among females) in parallel with inactivity. Median age at smoking initiation was decreasing rapidly from 18 to 20 years in young adults to 15 in adolescents. A statistically significant social gradient in disfavor of the lower education level was observed for overweight in all age groups of women above 16 (odds ratios (ORs) 2.4 to 3.3, P < 0.01), for inactivity in adult males (ORs 1.6 to 2.0, P < 0.05), and for regular smoking in older adolescents (OR 1.9 for males, 2.7 for females, P < 0.005), but not for elevated blood pressure. CONCLUSION: Discontinuities in the cross-sectional age patterns of CRF indicated the emergence of a social gradient and the need for preventive actions against the early adoption of persistent unhealthy behaviors, to which low-educated girls and women are particularly exposed. PMID- 12590998 TI - The association between high blood pressure, physical fitness, and body mass index in adolescents. AB - BACKGROUND: The aim of this study was to analyze the association of fitness and fatness with blood pressure (BP) and hypertension. This was a cross-sectional study of 13,557 boys and girls 15-20 years of age. Fitness was estimated from a shuttle run test, fatness from body mass index (BMI), and BP was measured sitting after 5 min of rest. Other lifestyle variables were self-reported. RESULTS: Boys had a higher systolic BP (SBP) than girls. A low physical fitness level and high BMI were independently associated with a high BP and risk of having hypertension in both girls and boys. Interaction was found between BMI and fitness. In a stratified analysis an odds ratio (OR) of 3.99 was found for hypertension in girls with a BMI > 25 kg m(-2) compared to lean girls if all had a low fitness level, and an OR of 2.14 for a high BMI in girls with a high fitness level. In boys, OR for high versus low BMI were 3.23 in the low fit and 2.34 and 2.50 in the middle and upper tertile of fitness, respectively. CONCLUSIONS: Fitness and BMI were independently associated to BP. BMI was a stronger predictor of hypertension in those with a low fitness level, especially in girls. PMID- 12590999 TI - An environmental intervention to promote walking and cycling--the impact of a newly constructed Rail Trail in Western Sydney. AB - BACKGROUND: Few studies have evaluated environmental changes to promote physical activity. The impact of a local promotional campaign around a newly constructed Rail Trail in western Sydney, Australia, was assessed. METHODS: Pre and post campaign telephone surveys were used. Adults 18-55 years of age were randomly sampled from an "inner" area, residents within < or =1.5 km of the Trail, and an "outer" area, bike-owners only, 1.5-5 km from the Trail. Objective concurrent monitoring of daily bike counts was carried out. RESULTS: A cohort of 450 completed both interviews. A significant increase (+2.9%, McNemar P < 0.05) in unprompted Trail awareness was detected but post-campaign awareness was low (34%). Inner cyclists, males, and those recalling any baseline bike promotion messages were more likely to be aware of the Trail. Trail usage was higher among bike-owners than pedestrians (8.9 vs 3.3%, P = 0.014) and was moderated by proximity to the Trail. Inner cyclists increased mean cycling time by 0.19 h (SD = 1.5) while outer cyclists decreased cycling time (-0.24 h, SD = 1.6). Mean daily bike counts in the monitored areas increased significantly after the Trail launch (OR = 1.35, P = 0.0001, and OR = 1.23, P = 0.0004). CONCLUSIONS: The campaign reached and influenced cyclists in the inner area. For further increase in Trail usage promotional and educational programs focusing on walking for inner residents are needed. PMID- 12591000 TI - Glucocorticoid-induced osteoporosis: pathogenesis, diagnosis, and management. AB - Glucocorticoid-induced bone loss is dose- and duration-related, develops rapidly (within months of therapy), and leads to an increased risk of fractures. Moreover, less than one in four patients prescribed oral glucocorticoids receive any treatment to prevent or treat osteoporosis. The American College of Rheumatology recommends bisphosphonate therapy to prevent bone loss in most patients beginning long-term glucocorticoid therapy (prednisone equivalent of > or =5 mg/day for at least 3 months), and in men and postmenopausal women receiving long-term glucocorticoids who have an abnormal bone mineral density (T score below -1). Patients with glucocorticoid-induced osteoporosis are at particularly high risk for fractures, and should be treated aggressively to reduce fracture risk. Risedronate is approved in the United States for both prevention and treatment of glucocorticoid-induced osteoporosis and alendronate is approved for treatment. Both drugs increase bone mass in patients with established glucocorticoid-induced osteoporosis. Risedronate has been shown to significantly reduce the incidence of fractures after 1 year of treatment. Prevention or treatment of glucocorticoid-induced bone loss is recommended for patients at risk. PMID- 12591001 TI - Research vs. public health practice: when does a study require IRB review? PMID- 12591002 TI - Effects of nicotine on thromboxane/prostacyclin balance in myocardial ischemia. AB - It has been proven that nicotine contributes to cardiovascular diseases, although its precise mechanism of action is still unclear. The purpose of this study is to find how nicotine may complicate myocardial ischemia by affecting the thromboxane/prostacyclin (TXA(2)/PGI(2)) balance. We used four groups (n=7 each) of isolated and perfused rabbit hearts according to Langendorff method: (i) control group; (ii) group submitted to 1 microM nicotine perfusion during 60 min; (iii) group submitted to a regional ischemia by ligation of the left descending coronary artery during 60 min and (iv) group submitted to nicotine perfusion during ischemia. Levels of TXB(2) and 6-keto PGF(1alpha), the stable metabolites of TXA(2) and PGI(2) were then determined in the microsomes of the hearts by radioimmunoassay. The results showed that (1) a TXA(2) synthetase activity is present in the myocardium, and this activity, as well as that of PGI(2) synthetase, is decreased by a 60min ischemia; (2) TXA(2) and PGI(2) activities are not affected by nicotine in the normal myocardium and (3) nicotine infusion during ischemia contributes to the increase of TXA(2)/PGI(2) ratio further by decreasing PGI(2). Therefore, these results provide one explanation on how nicotine might worsen myocardial ischemia. PMID- 12591003 TI - Influences of prostanoids and nitric oxide on post-suspension hypotension in female Sprague-Dawley rats. AB - Impairment in cardiovascular functions sometimes manifested in astronauts during standing postflight, may be related to the diminished autonomic function and/or excessive production of endothelium-dependent relaxing factors. In the present study, using the 30 degrees head-down tilt (HDT) model, we compared the cardiovascular and biochemical effects of 7 days of suspension and a subsequent 6 h post-suspension period between suspended and non-suspended conscious female Sprague-Dawley rats. Mean arterial pressure (MAP) and heart rate were measured prior to suspension (basal), daily thereafter, and every 2h post-suspension. Following 7 days of suspension, MAP was not different from their basal values, however, upon release from suspension, MAP was significantly reduced compared to the non-suspended rats. Nitric oxide levels were elevated while thromboxane A(2) levels declined significantly in both plasma and tissue samples following post suspension. The levels of prostacyclin following post-suspension remained unaltered in plasma and aortic rings but was significantly elevated in carotid arterial rings. Therefore, the post-suspension reduction in mean arterial pressure is due mostly to overproduction of nitric oxide and to a lesser extent prostacyclin. PMID- 12591004 TI - Bioavailability of omega-3 essential fatty acids from perilla seed oil. AB - Increased dietary intake of fish oil omega-3 fatty acids, eicosapentanoic acid and docosohexanoic acid, and their precursor, alpha-linolenic acid (ALA), is associated with various health benefits. Enteric-coating (Entrox), which improves stability of omega-3 capsules, has been shown to facilitate fish oil absorption after chronic treatment. To assess the effect of Entrox coating on the short-term bioavailability of ALA administered in the form of ALA-rich Perilla seed oil, 12 healthy subjects (6 males and 6 females) received in a random order Entrox-coated and non-coated ALA formulations, each as a single 6g dose separated by a 3-week washout period. Measurements of plasma ALA concentrations from 0 to 24h showed no difference in ALA pharmacokinetics between the two formulations. However, significantly greater increases in plasma ALA levels from baseline to 24h were observed after ingestion of Entrox vs. non-coated product, suggesting a possible benefit of Entrox with long-term treatment. PMID- 12591005 TI - Pilot study of dietary fatty acid supplementation in the treatment of adult periodontitis. AB - The anti-inflammatory effects of both n-3 and n-6 polyunsaturated fatty acids (PUFA) have been demonstrated in vitro and in many disease states, in particular in the treatment of rheumatoid arthritis. The benefit of n-3 PUFA supplementation has been documented in animal models of periodontal inflammation and a trend towards reduced inflammation has been seen in human experimental gingivitis. The purpose of this study was to examine the potential anti-inflammatory effects of PUFA supplementation, by administration of fish oil as a source of the n-3 PUFA, eicosapentaenoic acid, and borage oil as a source of the n-6 PUFA, gamma linolenic acid (GLA), to adults with periodontitis. Thirty adult human subjects with periodontitis were administered either fish oil 3000 mg daily; borage oil 3000 mg daily; fish oil 1500 and borage oil 1500 mg daily, or placebo. The modified gingival index, the plaque index (PI), periodontal probing depths and beta-glucuronidase levels in gingival crevicular fluid were measured at baseline and after 12 weeks of treatment. Improvement in gingival inflammation was observed in subjects treated with borage oil (P<0.016), with a trend apparent in subjects treated with fish oil or a combination of PUFA. There was no statistically significant improvement in PI, although a trend was apparent in those receiving borage oil. Improvement in probing depth was seen in those subjects treated with either fish oil alone or borage oil alone, but statistical significance was only seen for the comparison of borage oil and placebo (P<0.044). No change was seen in gingival crevicular fluid (GCF) beta glucuronidase levels. The use of borage oil supplementation, a source of the n-6 PUFA, GLA, can have beneficial effects on periodontal inflammation. n-6 PUFA supplementation seemed to offer more impressive results than either n-3 PUFA supplementation or the combination of lower doses of the two supplements. Additional studies will be necessary to more fully assess the potential of these agents to favorably affect periodontal inflammation. PMID- 12591006 TI - Could n-3 polyunsaturated fatty acids reduce pathological pain by direct actions on the nervous system? AB - The intake of n-3 polyunsaturated fatty acids (PUFAs) in many industrialized countries is relatively low and its increased consumption has protective and modifying effects on such diverse conditions as atherosclerosis, ventricular arrhythmias, multiple sclerosis, major depression and inflammatory and autoimmune diseases. In addition, n-3 PUFAs have been shown to alleviate pain in patients with rheumatoid arthritis, inflammatory bowel disease and in a number of other painful conditions. This has been attributed to the inhibition of pro inflammatory eicosanoid and cytokine production by peripheral tissues. n-3 PUFAs have also been shown to inhibit eicosanoid production in glial cells, block voltage-gated sodium channels (VGSCs), inhibit neuronal protein kinases and modulate gene expression. They also appear to have mood-stabilizing and sympatholytic effects. The present article explores the possibility that, based on what is known about their neural and non-neural effects, n-3 PUFAs directly attenuate the neuronal and glial processes that underlie neuropathic and inflammatory pain. PMID- 12591007 TI - Elevated levels of endothelin-1 and prostaglandin E2 and their effect on nitric oxide generation in placental tissue from neonatal streptozotocin-induced diabetic rats. AB - Endothelin-1 (ET-1), nitric oxide (NO) and prostaglandin E(2) (PGE(2)) are regulators of feto-placental hemodynamics. In this study we explore the inter regulatory pathways that modulate the levels of these vasoactive agents in control and neonatal streptozotocin-induced (n-stz) diabetic rat placenta. ET-1 levels are increased in diabetic placenta when compared to controls (P<0.001), and are strongly reduced by an NO synthase inhibitor (P<0.001). PGE(2) production is increased in diabetic placenta when compared to controls (P<0.01), but these levels are not modulated by ET-1. NO levels, similar in control and in diabetic placenta, are not influenced by PGE(2), but they are negatively modulated by ET-1 in both control (P<0.05) and diabetic (P<0.01) placenta. We conclude that rat placental ET-1 inhibits NO levels but does not modify PGE(2) concentrations. The elevated levels of ET-1 and PGE(2) in diabetic placenta, potent vasoconstrictors of placental vasculature, are probably related to the induction of placental insufficiency and fetal hypoxia in this pathology. PMID- 12591008 TI - Role of metoprolol, B1-adrenoceptor antagonist, thromboxane A2 and nitric oxide in CsA-induced hypertension. AB - Chronic treatment with cyclosporine A (CsA), a potent immunosuppressive agent, is associated with the development of arterial hypertension. The effect of CsA on vascular responses was determined in Sprague-Dawley rats and rat aortic rings. Male rats weighing 250-300 g were given either CsA (25mg/kg/day) in olive oil or vehicle by intraperitoneal (ip) injection for 7 days. CsA administration produced a 42% increase (P<0.001) in mean arterial pressure (MAP) which reached a plateau after 3 days. The level of both nitrate/nitrite (NO(2)/NO(3)), metabolites of nitric oxide (NO), decreased by 50% (P<0.001), but the level of thromboxane A2 (TBXA2) increased by 75% (P<0.001), in the urine. When 10(-9)M of CsA was added acutely to intact aortic rings from untreated rats, NO(2)/NO(3) production decreased by 83% (P<0.011), but TBXA2 production increased by 86% (P<0.001). The effects of CsA were reversed both in vivo and in vitro by pretreatment with metoprolol (15 mg/kg/day ip), B1-adrenoceptor antagonist. There were no changes in MAP and tension in rats treated with metoprolol alone. In addition, in aorta of rats that were treated with CsA ip for 7 days, CsA significantly activated protein kinase C (PKC) translocation. This suggests that PKC mediate, in part, CsA-induced hypertension. In summary, CsA inhibits endothelial NO formation, activate PKC, and increase TBXA2 production, with resulting increase in MAP, and this changes can be overcome by pretreatment with metoprolol. PMID- 12591009 TI - Effects of sphingosine-1-phosphate and lysophosphatidic acid on human osteoblastic cells. AB - The effects of the lysophospholipids, sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) were studied in human primary osteoblastic cells and the human osteosarcomal cell lines, G292 and MG-63. The studies focused on the role of the Gi protein in the regulation of S1P and LPA-induced proliferation, the effects of the phospholipids on alkaline phosphatase, an early marker of osteoblastic cell proliferation, and the presence of edg receptors. Proliferation was assessed by 3H-thymidine incorporation. Short-term incubation with S1P or LPA induced increases in proliferation that were attenuated in the presence of the Gi inhibitor, pertussis toxin. Alkaline phosphatase activity was measured with a spectrophotometric assay. Biphasic effects of S1P and LPA were observed with the nature of the response dependent upon the cell type, concentration of test agent and the time period of incubation. RTPCR studies revealed that edg-1,2,4,5 receptors are present in the primary normal osteoblastic cells, the MG63 and G292 cells. Only the G292 cells expressed the edg-3 receptor to any significant extent. PMID- 12591010 TI - Effect of atorvastatin and clopidogrel on cellular immune function. AB - Lipid lowering therapy by statins and antiaggregation have become the basis of any anti-atherosclerotic prophylaxis either as primary or secondary prophylaxis. As several recent papers indicated immunosuppressive properties of statins we investigated changes in lymphocyte subpopulations, apoptosis markers, and cellular immune response towards mitogens after a short-term therapy with atorvastatin and clopidogrel. Nine healthy volunteers (four male, five female, age ranging from 26 to 43 years) were treated with 20 mg atorvastatin for 4 weeks and for 2 additional weeks with 20 mg atorvastatin and 75 mg clopidogrel after oral consent was given. Lymphocyte subpopulations were counted by flow cytometry. To assess cellular in vitro immune function, lymphocyte transformation tests with four mitogens (PHA, ConA, PWM, and OKT3) were performed. Absolute leucocyte counts remained unchanged as well as the granulocyte, monocyte, lymphocyte, and lymphocyte subpopulation counts. There were no detectable changes in markers of cell activation (HLA-DR, CD25, CD69, and CD86) or apoptosis (CD95, annexin). Cellular in vitro responses towards four mitogens did not show significant changes after atorvastatin nor after atorvastatin plus clopidogrel treatment.In conclusion, our data show that atorvastatin is not an immunosuppressive drug under therapeutical conditions. PMID- 12591011 TI - Effects of chronic alcohol consumption on myocardial ischemia in rats. AB - The effects of chronic alcohol consumption on myocardial ischemia and gas perfusion with 95% O(2)-5% CO(2) were investigated in isolated rat heart. Eighteen adult male Wistar rats were used. Rats were assigned into six groups for each group to contain three rats: normal, alcoholic, normal ischemic, alcoholic ischemic, normal ischemic and 95% O(2)-5% CO(2) perfused, alcoholic ischemic and 95% O(2)-5% CO(2) perfused, respectively. Alcohol (7.2%, v/v) was given to rats by a modified liquid diet for 21 days. Rats were anaesthetized with ketamine (1 2mg kg(-1)). Hearts were quickly isolated. Normal and alcoholic rat hearts were directly sent to the electron microscopic preparation. The other hearts were cut into small pieces and put into Krebs solution. The solution was continuously bubbled using 95% N(2)-5% CO(2) 20 min for ischemia. After removal of normal ischemic and alcoholic ischemic heart specimens for electron microscopic examination, the remaining hearts of the last two groups were bubbled with 95% O(2)-5% CO(2) for another 20 min for the purpose of reperfusion and then were also prepared for electron microscopic examination. The hearts were investigated with a transmission electron microscope (Jeol 100 CXII TEM). Twenty-one days of chronic alcohol consumption was found to have no significant effect on myocardial ischemia determined by transmission electron microscopic examination. Our results suggest that there is no significant relationship between 21 days of alcohol consumption by a liquid diet and myocardial protection. PMID- 12591012 TI - In vivo and in vitro pharmacodynamic properties of metoprolol in aortic coarctated rats. AB - The chronotropic effects of metoprolol (MP) in vivo after intravenous administration and in vitro in isolated atria of aortic coarctated rats were studied. MP levels in arterial dialysates and changes in arterial pressure and heart rate (HR) were determined after intravenous administration of MP. Affinity of MP by means of shifting of noradrenaline concentration-response curve and the inverse agonist activity of MP were studied in isolated atria. The IC(50) value of MP cardiac effect was smaller in aortic coarctated rats (13.0+/-5.4 ng ml(-1), n=5) than in sham operated (SO) rats (36.2+/-4.6 ng ml(-1), n=5). In isolated atria, no difference was found in MP dissociation constant and its inverse agonist activity between both experimental groups. In conclusion, the affinity and inverse agonism activity of MP would not be altered in aortic coarctated rats. A greater sensibility to the chronotropic effect of MP was found in coarctated rats. PMID- 12591013 TI - Neuropathology of aluminum toxicity in rats (glutamate and GABA impairment). AB - To get better insights into the pathological and biochemical defects underlying aluminum toxicity in brain tissue, we exposed male albino rats for 35 days to aluminum sulfate by gavage. Tissue aluminum level of brain was assessed, histological sections of brain were examined and amino acid transmitters contents were detected by reversed phase high performance liquid chromatography. Aluminum levels were high in brain specimens of the treated groups comparing to the control and it was dose-dependent. Marked increase in glutamate and glutamine levels was noticed while GABA level was significantly decreased. The most pronounced changes in brain tissue included spongioform changes in the neurons specially those of hippocampus, nuclear deformity, and neurofibrillary degeneration, similar to neurofibrillary tangles in Alzheimer's disease. It is concluded that accumulated aluminum in brain and altered amino acid neurotransmitters are important mechanisms of aluminum neurotoxicity. PMID- 12591014 TI - Effect of nutritive and tonic crude drugs on physical fatigue-induced stress models in mice. AB - The present study was undertaken to investigate the acute anti-fatigue effect of a liquid nutritive and tonic crude drugs (NTDs) on stress induced in mice. After forced walking for 3 or 6h, the NTDs (applied orally, 10 ml/kg) significantly increased locomotor activity, while the administration of NTDs after rapid eye movement (REM) sleep deprivation stress and after immobilization stress did not show a specific effect, having a similar effect as the vehicle with added vitamins, taurine and caffeine. The administration of NTDs after freezing due to electric shock stress showed a specific effect which was not seen in other control groups, water, vehicle (ethanol) and vehicle including vitamins, taurine and caffeine and so resemble the specific effect of NTDs in the stress of forced walking. The present results indicate that the NTDs produced an anti-fatigue effect on the decreased locomotor activity after forced walking and immobility induced by electric stimulation. However, the crude drugs were not effective in improving immobility after sleep deprivation or immobilization stress. PMID- 12591015 TI - Alteration of local ACE activity and vascular responsiveness during development of 2K1C renovascular hypertension. AB - OBJECTIVES: This study sought to examine the correlation between development of hypertension and local, including; aorta, heart, kidney, lung, as well as circulatory (serum) ACE activity in two kidney one clip (2K1C) renovascular hypertension. METHODS: Ten- to twelve-week-old rats undertaken left renal artery clipping. Experiments were carried out in 2, 4, 8, and 12 weeks after induction of hypertension (2, 4, 8 and 12W). After sacrificing, animals blood and tissues including heart, aorta lung and kidney were dissected out and homogenized in Trizma buffer. ACE activity was determined by hydrolysis of the synthetic substrate, Hip-His-Leu and the amount of hippuric acid liberated from the substrate were analyzed by HPLC. Vascular responsiveness was measured using perfusion pressure method. RESULTS: The systolic blood pressure (SBP) was gradually increased in 2K1C animals and was markedly higher compared to that of controls. The ACE activity in all tissues from 2K1C was significantly different in all groups of 2, 4, 8, and 12 weeks after surgery. The serum ACE activity of 2K1C was markedly increased in 2 and 4W and reached to plateau in 8 and 12W group. Vascular responsiveness to angiotensin I (AngI) was increased during development of hypertension in all groups of animals. CONCLUSION: These results indicated that there is a positive correlation between augmentation of blood pressure and local ACE activity in various tissues as well as serum, highlighting the significant contribution of local compared to circulatory ACE activity in development of renovascular hypertension. PMID- 12591016 TI - Role of leptin on alcohol-induced oxidative stress in Swiss mice. AB - Previous studies suggest a possible link between leptin and hepatic inflammation; however, the role of leptin on liver disease remains unclear. The purpose of the present study was to evaluate the effect of leptin on tissue lipid peroxidation and the antioxidant status in experimental hepatotoxicity. Administering ethanol (6.32 g/kg body weight) to 4-week-old healthy mice for 45 days resulted in significantly elevated levels of tissue thiobarbituric acid reactive substances (TBARS), conjugated dienes (CD) and lowered activities of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and glutathione related enzymes such as glutathione peroxidase (GPx) and glutathione S-transferase (GST) as compared with those of the control mice. subsequent to the experimental induction of hepatotoxicity (i.e. after the initial period of 30 days) exogenous leptin was simultaneously administered (230 microg/kg body weight) every alternate day for 15 days along with the daily dose of alcohol. Leptin administration to control and alcohol-treated mice significantly reduced the weight gain, significantly elevated the liver and kidney levels of TBARS and CD, and significantly lowered the levels of enzymic and non-enzymic antioxidants as compared with the untreated control and alcohol supplemented mice. It is postulated that the increase in systemic leptin levels enhance the oxidative stress, and lower the antioxidant defence, leading to augmented hepatic inflammation in alcoholic liver disease. PMID- 12591017 TI - Contribution to characterization of oxidative stress in HIV/AIDS patients. AB - Infection by human immunodeficiency virus (HIV) causes persistent chronic inflammation. Viral Tat protein plays a role in the intracellular increase of reactive oxygen species (ROS) thus increasing apoptotic index, mostly the one mediated by FAS/CD95, and depleting CD4+ T lymphocytes. The aim of this study was to investigate whether there is a relationship between an extensive array of redox status indices (glutathione (GSH), malondialdehyde (MDA), peroxidation potential, total antioxidant status, glutathione peroxidase (GPx), superoxide dismutase (SOD), total hydroperoxide (TH), DNA fragmentation) and relative CD4, CD95, CD38/CD8 T lymphocyte counts in HIV/AIDS patients compared to healthy subjects. Blood samples from 85 HIV/AIDS patients and 40 healthy subjects were tested by spectrophotometric techniques in order to measure oxidative stress indices, and by flow cytometry to quantify T cell subsets. Patients were divided in two groups according to CDC 1993 guidelines. CD95 and CD38 increase paralleled the severity of HIV infection. Both a reduction of GSH levels and an increase in MDA and TH levels were detected in the plasma of HIV+ patients. These patients also showed an increase of DNA fragmentation in lymphocytes as well as a significant (P<0.05) reduction of GPx and an increase in SOD activity in erythrocytes. Relatively to the control group, HIV-infected patients had significantly differences in global indices of total antioxidant status. These results corroborate that substantial oxidative stress occurs during HIV infection. To our knowledge this study is the first relating oxidative stress indices with both CD38/CD8 and CD95 lymphocytes subsets. PMID- 12591018 TI - Anseculin improves passive avoidance learning of aged mice. AB - Anseculin is a new coumarine derivative with promising cognition improving properties. We investigated its possible effects on passive avoidance learning in young and aged mice and the possible relationship to its alpha(1)-antagonistic properties.A single dose of anseculin did not affect long-term memory of young and aged mice. Moreover, in young mice, subchronic treatment with anseculin had no effect on cognitive functions, whereas aged, cognitively impaired mice clearly showed improved long-term memory with no effects on short-term memory. Further biochemical studies of aged control mice showed in comparison to the young mice a significant decrease of central alpha(1)-adrenoceptors density, but unchanged alpha(1)-adrenoceptor-mediated IP hydrolysis. Subchronic treatment with anseculin had no effect on the density of central alpha(1)-adrenoceptors in young adult mice and only a small although not significant effect on the density of cognitively-impaired aged mice. Furthermore, anseculin has no effects on alpha(1) adrenoceptor-mediated IP hydrolysis. PMID- 12591019 TI - Reversible conduction block in isolated frog sciatic nerve by high concentration of bupivacaine. AB - We evaluated the effects of a high concentration of bupivacaine commonly used for spinal anaesthesia on the reversibility of conduction block and compound nerve action potential (CNAP) parameters in isolated frog sciatic nerve measured by extracellular recording technique. Isolated frog sciatic nerves were bathed in 1.3% bupivacaine solution for 20 min. In each nerve, action potentials were recorded before exposure to bupivacaine solution, which served as the control data. The extracellular action potentials were recorded after 20 min in the drug by using a BIOPAC MP 100 Acquisition System Version 3.5.7 (Santa Barbara, USA). The nerves were washed for 3h continuously with Ringer's solution and action potentials were recorded. The nerve was then soaked overnight in Ringer's solution at room temperature and tested for impulse recovery. There were significant differences among the experiments regarding CNAP peak-to-peak amplitude, area and duration but conduction velocities among the experiments did not show any statistical difference. In the presence of bupivacaine the extracellular action potential amplitude decreased by 46.99+/-29.31% relative to the control amplitude (P<0.05), recovered to 47.10+/-26.90% after 3h of wash, and reached 123.20+/-39.70% after the overnight soak process. This study showed that exposing nerve to high concentration of bupivacaine causes reversible impulse blockade and that bupivacaine does not cause neurotoxic effect on isolated frog sciatic nerve. PMID- 12591020 TI - Progression of renal failure with anaemia and multiple effects of angiotensin converting enzyme inhibitor in rats with renal mass reduction. AB - Several factors such as proteinuria and renal fibrosis may be important in the progression of many forms of chronic renal diseases. The purposes of the current study were to investigate the progressive renal failure of the rats with surgical renal mass reduction (RMR) and the effect of the angiotensin-converting enzyme (ACE) inhibitor, lisinopril, and to document correlation of several factors associated with progressive renal failure. Rats were subtotal (5/6) nephrectomized by resection of the renal poles and sham-operated. The functional, histological and haematological changes of the rats were studied for up to 10 weeks. After 2 weeks of RMR, oral administration of lisinopril (10 mg kg(-1) per day) was performed for 8 weeks. RMR resulted in progressive renal failure with proteinuria, monocyte/macrophage (ED1+) infiltration, anaemia as assessed by haemoglobin and haematocrit (Htc), renal hypertrophy as assessed by left kidney to body weight ratio (BKW/BW), and renal fibrosis as assessed by glomerular lesions and tubulointerstitial changes. Lisinopril exhibited renoprotection with antiproteinuric effect and inhibition of monocyte/macrophage (ED1+) infiltration. However, beneficial effect of lisinopril on anaemia was not observed. At 10 weeks after surgery, severity of proteinuria positively correlated with plasma creatinine (Pcr), BKW/BW, histological damage, and systolic blood pressure, and negatively correlated with haemoglobin. Severity of tubulointerstitial changes positively correlated with Pcr and blood urea nitrogen, and negatively correlated with haemoglobin and Htc. Moreover, monocyte/macrophage (ED1+) infiltration positively correlated with severity of proteinuria and tubulointerstitial changes. These findings strongly support that proteinuria, monocyte/macrophage infiltration and renal fibrosis appear to play principal roles in the progressive renal failure with anaemia and renoprotection of ACE inhibition may be mediated by multiple actions of ACE inhibitor. The present study confirms that rats with RMR is useful to explore target molecules for renoprotective drugs and evaluate renoprotective effect of new molecular entities. PMID- 12591021 TI - In vivo photosensitizing efficiency of a diphenylchlorin sensitizer: interest of a DMPC liposome formulation. AB - The pharmacokinetic behaviour and phototherapeutic effectiveness of 2,3-dihydro 5,15-di(3,5-dihydroxyphenyl)porphyrin (SIM01), a new diphenylchlorin photosensitizer incorporated into dimyristoyl phosphatidylcholine (DMPC) liposomes, were studied in vivo in nude mice bearing HT29 human adenocarcinoma. The photophysical and photochemical specificity of SIM01 are a strong absorption at 647 nm, high photosensitizing efficiency and a rapid pharmacokinetic profile making SIM01 an attractive candidate for PDT. The pharmacokinetics in vivo, as determined by fiber spectrofluorimetry, showed that tumor concentration was found maximal for SIM01 and SIM01-DMPC 12h after injection, with better uptake for the liposomal formulation. With a 2 mg kg(-1) dye dose, optimal PDT response occurred when the interval between injection and irradiation was 12h for both formulations, with laser irradiation of 300J cm(-2) (650 nm, 300 mW). At day 12 after treatment involving a 12-h interval between injection and irradiation, tumor growth was decreased by 26% for SIM01 (P=0.005) and 35% for SIM01-DMPC (P=0.001) as compared to the untreated group.SIM01 would appear to be a powerful sensitizer characterized by high in vivo phototoxicity and rapid tissue uptake and elimination. Our results suggest that SIM01 delivered in a liposomal dispersion is as effective as the raw formulation, something that would open new delivery routes and PDT applications. PMID- 12591022 TI - Using executive heterogeneity to explore the nature of working memory deficits in Parkinson's disease. AB - Idiopathic Parkinson's disease (IPD) is characterised by a triad of motor symptoms, namely bradykinesia, rigidity and resting tremor, although cognitive impairment is a common feature of the disease and has been accepted as one of the strong predictors of quality of life in such patients. Neuropsychological testing in Parkinson's disease often reveals a pattern of cognitive impairment similar to that observed in patients with frontal lobe lesions, although clear differences between the two groups have also often been reported. This apparent inconsistency in the literature may reflect heterogeneity among different groups of patients with Parkinson's disease, although to date this possibility has not been formally addressed. In this study, two groups of patients with Parkinson's disease were assessed on a novel verbal memory task, which allowed different aspects of working memory function such as maintenance, retrieval and manipulation to be tested within the same general paradigm. The two groups were selected according to either good or bad performance on a 'standard', visuospatial test of executive function (The Tower of London Task), but were well matched on all other demographic and cognitive measures tested. The sub-group of Parkinson's disease patients with Tower of London defined executive deficit were specifically impaired at manipulating information within verbal working memory, both compared to controls and to the group of patients with no predefined executive impairments. In contrast, the three groups did not differ in their ability to maintain or retrieve information within verbal working memory. Given the known preferential role of the dorsolateral frontal cortex in higher executive functions, (including both planning and the manipulation of information within working memory), these results suggest that, in a subset of Parkinson's disease patients, it is the frontostriatal circuitry involving this region which is primarily affected, while other components of this circuitry may be relatively spared. The results also suggest that performance on the Tower of London task may prove to be a useful discriminant variable in defining the nature of cognitive heterogeneity in Parkinson's disease. PMID- 12591023 TI - The prefrontal cortex and conscious monitoring of action: an experimental study. AB - To investigate the role of the prefrontal cortex in conscious monitoring, we used an experimental paradigm generating a conflict between the action planned and the sensory-motor feedback. We analyzed the acquisition of explicit knowledge of the strategy for resolving the conflict and its influence on motor adaptation. Twenty patients with frontal lobe lesions and 18 controls had to trace a sagittal line with a stylus on a graphics tablet. A mirror on which the traced line, processed by a computer, was projected hid the hand. A mask limited visual feedback to the last third of the trajectory. Without informing the subjects, the line traced was modified by introducing a bias of 24 degrees to the right. To succeed in the task, subjects had to modify their motor program and to deviate their trajectory in the opposite direction. Conscious elaboration of the strategy was evaluated by the number of trials needed to explicitly report the required deviation. Three groups of patients were distinguished: (1). with normal explicit strategy; (2). with delayed explicit strategy, and (3). without explicit strategy at the last trial. They significantly differed by the severity of the dysexecutive syndrome, particularly of environmental adherence. Motor adaptation was evaluated by the area between the line traced and the ideal line to compensate for the deviation. In patients with normal elaboration of the strategy, motor control was similar to that of controls, but it was severely disturbed in the other two groups. These results suggest the involvement of the prefrontal cortex in conscious motor monitoring. PMID- 12591024 TI - Significant relation between MR measures of planum temporale area and dichotic processing of syllables in dyslexic children. AB - In the present study, we investigated differences between dyslexic and normal reading children in asymmetry of the planum temporale area in the upper posterior part of the temporal lobe and dichotic listening performance to consonant-vowel syllables. The current study was an extension of previous studies in our laboratory on the same participants, now including also girls and left-handers. There were 20 boys and 3 girls in the dyslexic group and 19 boys and 4 girls in the normal reading group. The age of the participants was 10-12 years for both groups. The participants were screened from a population of 950 students in the fourth school grade in the greater Bergen district. The planum temporale area was measured in sagittal magnetic resonance (MR) images. Mean left and right area and asymmetry index were compared between the groups. Dichotic presentations of consonant-vowel syllables made it possible to separately probe left and right hemisphere phonological function, and to correlate this with planum temporale area. The results showed a significantly larger left than right planum temporale area for both groups. However, while the right planum temporale area was similar for the dyslexic and control groups, the left planum temporale was significantly (one-tailed t-test) smaller in the dyslexic group. Both groups also showed a significant right ear advantage to the consonant-vowel syllables in the dichotic listening test. The relation between planum temporale and dichotic listening asymmetry showed a significant correlation for the dyslexic group only, indicating a positive relation between brain structure and function in dyslexic children. The results are discussed in terms of important subject characteristics with regard to brain markers of dyslexia. PMID- 12591025 TI - Divided attention, selective attention and drawing: processing preferences in Williams syndrome are dependent on the task administered. AB - The visuo-spatial abilities of individuals with Williams syndrome (WS) have consistently been shown to be generally weak. These poor visuo-spatial abilities have been ascribed to a local processing bias by some [R. Rossen, E.S. Klima, U. Bellugi, A. Bihrle, W. Jones, Interaction between language and cognition: evidence from Williams syndrome, in: J. Beitchman, N. Cohen, M. Konstantareas, R. Tannock (Eds.), Language, Learning and Behaviour disorders: Developmental, Behavioural and Clinical Perspectives, Cambridge University Press, New York, 1996, pp. 367-392] and conversely, to a global processing bias by others [Psychol. Sci. 10 (1999) 453]. In this study, two identification versions and one drawing version of the Navon hierarchical processing task, a non-verbal task, were employed to investigate this apparent contradiction. The two identification tasks were administered to 21 individuals with WS, 21 typically developing individuals, matched by non-verbal ability, and 21 adult participants matched to the WS group by mean chronological age (CA). The third, drawing task was administered to the WS group and the typically developing (TD) controls only. It was hypothesised that the WS group would show differential processing biases depending on the type of processing the task was measuring. Results from two identification versions of the Navon task measuring divided and selective attention showed that the WS group experienced equal interference from global to local as from local to global levels, and did not show an advantage of one level over another. This pattern of performance was broadly comparable to that of the control groups. The third task, a drawing version of the Navon task, revealed that individuals with WS were significantly better at drawing the local form in comparison to the global figure, whereas the typically developing control group did not show a bias towards either level. In summary, this study demonstrates that individuals with WS do not have a local or a global processing bias when asked to identify stimuli, but do show a local bias in their drawing abilities. This contrast may explain the apparently contrasting findings from previous studies. PMID- 12591026 TI - Social cognition in frontotemporal dementia and Huntington's disease. AB - Frontotemporal dementia (FTD) and Huntington's disease (HD) are degenerative disorders, with predominant involvement, respectively of frontal neocortex and striatum. Both conditions give rise to altered social conduct and breakdown in interpersonal relationships, although the factors underlying these changes remain poorly defined. The study used tests of theory of mind (interpretation of cartoons and stories and judgement of preference based on eye gaze) to explore the ability of patients with FTD and HD to interpret social situations and ascribe mental states to others. Performance in the FTD group was severely impaired on all tasks, regardless of whether the test condition required attribution of a mental state. The HD group showed a milder impairment in cartoon and story interpretation, and normal preference judgements. Qualitative differences in performance were demonstrated between groups. FTD patients made more concrete, literal interpretations, whereas HD patients were more likely to misconstrue situations. The findings are interpreted as demonstrating impaired theory of mind in FTD, as one component of widespread executive deficits. In HD the evidence does not suggest a fundamental loss of theory of mind, but rather a tendency to draw faulty inferences from social situations. It is concluded that social breakdown in FTD and HD may have a different underlying basis and that the frontal neocortex and striatum have distinct contributions to social behaviour. PMID- 12591027 TI - Implicit face perception in a patient with visual agnosia? Evidence from behavioural and eye-tracking analyses. AB - This paper investigates face perception in a visual agnosic and prosopagnosic patient (SB). Despite very extensive lesions of visual areas, SB remains capable of some visual processing [Brain 125 (2002) 58]. However, in everyday situations SB does not exhibit signs of specific face recognition. To investigate how SB may process faces, we tested two hypotheses. According to the 'spared module hypothesis,' SBs abilities come from spared modules of implicit face processing. According to the 'general strategy hypothesis,' SB may have developed some deliberate compensatory strategies. A two-session experimental design was constructed. In both sessions, face and non-face pictures were shown to participants. In Session 1 (implicit condition), participants had to decide whether each picture was a vegetable. In Session 2 (explicit condition), participants had to decide whether each picture was a face. Verbal reports showed that SB was not aware of faces in Session 1. However, behavioural results showed that (1). SB could process faces; (2). even when SB was not aware of faces, he processed them differently than non-faces; (3). when knowing the presence of faces, he did not process faces better. In addition, eye-tracking data suggested that SB did not change the nature of his processing from Sessions 1 to 2. Pupil diameters showed that fixated facial features were processed similarly as in control participants. Together, these results are not compatible with a general compensatory strategy hypothesis and suggest sparing of an implicit face processing module in SB. PMID- 12591028 TI - An evaluation of two commonly used tests of unfamiliar face recognition. AB - The Warrington Recognition Memory for Faces (RMF) and the Benton Facial Recognition Test (BFRT) are commercially available tests that are commonly used by clinicians and cognitive neuropsychologists to evaluate unfamiliar face recognition. Yet, it is not clear that a normal score on either instrument demonstrates normal unfamiliar face recognition. Because the RMFs stimuli contain abundant non-internal facial feature information, subjects may be able to score in the normal range without using internal facial features. On the BFRT, subjects commonly rely on feature matching strategies using the hairline and eyebrows rather than recognizing the facial configuration. To test whether these routes to recognition can support normal performance, normal subjects were tested with versions of the RMF and the BFRT in which the faces had been painted over in a way that prevented the operation of some of the procedures normally involved with face recognition. Even though these modifications removed all of the internal feature information in the RMF, many subjects scored in the normal range, and despite precluding the use of configural processing in the BFRT, many of the scores were in the normal range. As a result, it is apparent that normal scores on these tests do not demonstrate normal unfamiliar face recognition and so clinicians should be cautious in interpreting scores in the normal range. Finally, these results place in question models supported by dissociations involving normal performance on these tests. PMID- 12591029 TI - Priming of strong semantic relations in the left and right visual fields: a time course investigation. AB - Prior time-course investigations of cerebral asymmetries in word processing have sometimes reported hemisphere differences in the onset and duration of semantic priming. In the current study, very strongly related word pairs (categorical associates such as arm-leg) were employed in a low relatedness proportion lexical decision priming paradigm. A range of prime-target stimulus onset asynchronies (SOAs: 150-800 ms) was included. Only very weak evidence was obtained for a LVF priming lag at the briefest SOA, while priming was bilateral at moderately long SOAs. We consider these data in the context of previous time-course studies and suggest that, when highly semantically similar word pairs are used, a right hemisphere priming lag is, at best, a very small effect. PMID- 12591030 TI - Facial expression recognition in Williams syndrome. AB - Individuals with Williams syndrome (WS) excel in face recognition and show both a remarkable concern for social stimuli and a linguistic capacity for, in particular, emotionally referenced language. The animated full facial expression comprehension test (AFFECT), a new test of emotional expression perception, was used to compare participants with WS with both chronological and mental age matched controls. It was found that expression recognition in WS was worse than that of chronologically age-matched controls but indistinguishable from that of mental age controls. Different processing strategies are thought to underlie the similar performance of individuals with WS and mental age controls. The expression recognition performance of individuals with WS did not correlate with age, but was instead found to correlate with IQ. This is compared to earlier findings, replicated here, that face recognition performance on the Benton test correlates with age and not IQ. The results of the Benton test have been explained in terms of individuals with WS being good at face recognition; since a piecemeal strategy can be used, this strategy is improved with practice which would explain the correlation with age. We propose that poor expression recognition of the individuals with WS is due to a lack of configural ability since changes in the configuration of the face are an important part of expressions. Furthermore, these reduced configural abilities may be due to abnormal neuronal development and are thus fixed from an early age. PMID- 12591031 TI - Visuo-motor control of the ipsilateral hand: evidence from right brain-damaged patients. AB - We investigated the extent to which the right hemisphere is involved in the control of the ipsilateral hand by analysing the kinematics of right-hand prehension in right brain-damaged (RBD) patients. We required patients to grasp one of five possible objects, equally-sized and distributed over a 40 degrees wide workspace. With the purpose of investigating the right hemisphere contribution to the on-line visuo-motor control, we also assessed patients' ability to correct their movement "in-flight", in response to a sudden change of object position. Patients' performance was compared to that of aged-matched controls. A Younger group of healthy subjects, matching the population classically tested on double-step paradigms, was also evaluated to fully assess whether patients' kinematics could be partially due to normal ageing. As a further aim, the possible influence of hemispatial neglect was evaluated by comparing the performances of right brain-damaged patients with and without neglect. In normal subjects, the results confirmed and extended the notion of (a). positional tuning of grip formation, and (b). fast reactions following a change in object position. In addition, subtle effects of ageing on visuo-motor behaviour were shown by less efficient movement correction in the Elderly group. Patients executing reach-to-grasp actions into the left contralesional hemispace were selectively affected in both temporal and spatial aspects of movements. While their performances were relatively well preserved in the right hemispace, patients did not show positional tuning of grip formation, nor fast corrections of their movements when acting in the left hemispace. Interestingly, similar deficits were found irrespective of the presence of neglect. These results show that the right hemisphere contributes to the processing of visuo-motor information that is necessary for executing actions with the ipsilateral hand in the contralateral space. PMID- 12591033 TI - ERCP during pregnancy. PMID- 12591034 TI - Estimating the importance of infection in IBS. PMID- 12591035 TI - Positive markers in AMA-negative PBC. PMID- 12591036 TI - Is type II diabetes another extrahepatic manifestation of HCV infection? PMID- 12591037 TI - Endoscopic, deep mural implantation of Enteryx for the treatment of GERD: 6-month follow-up of a multicenter trial. AB - OBJECTIVES: This prospective, multicenter, single-arm study evaluated the safety and efficacy of the endoscopic implantation of Enteryx, a biocompatible, non biodegradable liquid polymer for the treatment of GERD. METHODS: Eighty-five patients with heartburn symptoms responsive to proton pump inhibitor (PPI) use were enrolled. Inclusion requirements were HRQL score < or = 11 on PPI and > or = 20 off PPI, and 24-hour PH probe with > or = 5% total time at PH < or = 4. Patients with a hiatus hernia > 3 cm, grade 3 or 4 esophagitis, or esophageal motility disorder were excluded. Using a 4-mm needle tipped catheter during standard endoscopy, implants were made in 3-4 quadrants deep into the wall of the cardia. Use of PPI medications, pH-metry, manometry, GERD symptoms, and patient quality of life were assessed over a 6-month follow-up period. RESULTS: At 6 months, PPI use was eliminated in 74% and reduced by > 50% in 10% of patients. The median HRQL score improved from 24.0 pre-implant (baseline off PPIs) to 4.0 at 6 months (p < 0.001). Mean total esophageal acid exposure time was 9.5% pretherapy and 6.7% at 6 months (p < 0.001). Mean LES length increased from 2.0 cm at baseline to 3.0 cm posttherapy (p = 0.003). There were no clinically serious adverse events. Transient mild-to-moderate chest pain commonly occurred after implantation. CONCLUSIONS: The endoscopic implantation of Enteryx is a safe and effective therapy for eliminating or decreasing the need for PPI medications, improving GERD symptoms and patient quality of life, and decreasing esophageal acid exposure among patients suffering from GERD. PMID- 12591038 TI - Oral erythromycin and symptomatic relief of gastroparesis: a systematic review. AB - OBJECTIVE: Erythromycin is a motilin agonist that greatly increases the fractional rate of gastric emptying. Although a number of studies document the efficacy of erythromycin in improving gastric emptying, little information exists concerning symptom improvement in patients with gastroparesis. The aim of this study was to review clinical trials of erythromycin to determine the efficacy of this agent in producing symptom relief in patients with gastroparesis. METHODS: A MEDLINE search from 1966 to 2001 was performed to identify all clinical trials using erythromycin in patients with gastroparesis. The search was further limited to clinical trials using symptom assessment as an endpoint. References from index citations were reviewed to identify additional studies. The search was conducted independently by two authors, and discrepancies were resolved by consensus opinion. RESULTS: Thirty-five clinical trials were identified, and five met inclusion criteria. One study each involved gastroparesis caused by surgery and systemic sclerosis. Three studies evaluated patients with diabetic or idiopathic gastroparesis. No study used symptoms as a primary endpoint. Improvement was reported in 26 of 60 (43%) patients. Individual symptom scores were available for 23 of 60 subjects in these studies, and symptom improvement was seen in 11 of 23 (48%) patients. One study compared erythromycin and metoclopromide in an open label, crossover fashion, and found no difference between the two agents. All studies were methodologically weak and highly subject to bias. Four of five studies were open-label trials. Sample sizes in all studies were < or =13 subjects, and treatment duration was < or =4 wk in all studies. CONCLUSIONS: Although clearly a potent prokinetic, limited data exist concerning efficacy of erythromycin in treating gastroparesis. Small sample sizes, uncontrolled designs, short duration, and inadequate symptom assessment limit available studies. Well designed trials designed to assess symptom relief in gastroparesis are needed. PMID- 12591039 TI - Diabetic gastropathy and prokinetics. AB - The treatment of diabetic gastropathy, which here refers to a clinical syndrome of upper GI tract symptoms suggestive of an upper motility disturbance in diabetes whether or not there is delayed gastric emptying, remains suboptimal. New prokinetics and other motility-modifying agents may prove useful, but adequate clinical trials will be required to establish a role for them. However, diabetic gastropathy seems to represent a heterogenous syndrome in terms of pathophysiology, which potentially complicates the design of new randomized, controlled trials. This review aims to provide guidelines for future trials in this field. The evidence that delayed gastric emptying is a cause of symptoms in diabetic gastropathy is critically evaluated. The trial evidence supporting the short and long term efficacy of prokinetics is reviewed. Based on the available literature, it is concluded that improvement in gastric emptying does not equate with symptom relief in diabetic gastropathy. It is suggested that although gastric emptying should still be measured in clinical trials, it should not represent the primary outcome. The withdrawal treatment design applied in studies of diabetic gastropathy might be suboptimal. Double blind, parallel group studies remain the trial design of choice, but incorporation of validated outcome assessments and measurement of potential confounders of treatment response need attention in future trials. PMID- 12591040 TI - The percutaneous endoscopic gastrostomy tube. medical and ethical issues in placement. AB - OBJECTIVE: Offering and recommending PEG tube placement to patients has been a topic of considerable interest in the medical literature. The role of individual health care professionals in the decision making process is poorly defined. PEG tubes are often placed inappropriately because of unrealistic and inaccurate expectations of what they can accomplish in patients unable to tolerate adequate oral intake. We have developed an algorithm for PEG placement for the geriatric, oncology, and neurology patients based on a critical review of current literature. METHODS: An extensive review of the literature was performed focusing on PEG tube placement in oncology, neurology, and geriatric patients. This algorithm was developed to provide both the primary care provider and the specialist with appropriate indications for PEG placement in these patient populations. RESULTS: Appropriate indications for PEG placement are 1) Esophageal obstruction (e.g., esophageal cancer), 2) Neurologic etiology of dysphagia without obstruction (e.g., status post cerebrovascular accident, pseudobulbar palsy), 3) Prolonged refusal to swallow without evidence of concomitant terminal illness (e.g., protracted pseudodementia due to severe depression), 4) Supplemental nutrition for patients undergoing chemotherapy or radiation therapy. CONCLUSIONS: If no physiologic benefit is expected with PEG placement (anorexia cachexia syndrome), the health care team has no obligation to offer or perform an intervention. This same principle would apply if intervention improves physiologic states but has no effect on quality of life (e.g., permanent vegetative state). Small-bore feeding tubes are cost effective and relatively safe for enteral feedings of up to 6-8 weeks. This is especially pertinent in the population with acute neurological deficits, in which prognostication on extent of impairment is best estimated by communication with neurologist. In the geriatric population there is no proved benefit in weight gain or markers of nutrition (albumin, prealbumin) in patients with malnutrition due to impaired oral intake. PMID- 12591041 TI - Correlation between clinical severity score and the lower esophageal sphincter relaxation pressure in idiopathic achalasia. AB - OBJECTIVES: Achalasia is an esophageal motor disorder characterized by aperistalsis and incomplete relaxation of the lower esophageal sphincter (LES). The meaningful correlation between LES relaxation pressure and the severity of clinical symptoms is uncertain. The aim of this study was to elucidate the correlation between the clinical scoring and the LES relaxation pressure. METHODS: Over a 4-yr period from 1997 to 2001, all newly diagnosed patients with idiopathic achalasia were consecutively enrolled in a study. Diagnosis was established based on clinical, radiographic, endoscopic, and manometric criteria. The severity of five cardinal symptoms was scored on a scale of 0-3, and each patient received a total symptom score of 1-15. Manometry was subsequently performed, and the mean of five complete pull-through measurements was recorded as the resting LES relaxation pressure. RESULTS: A total of 115 patients (67 male and 48 female) with a mean age of 37.7 yr (range 12-90 yr) were included in the study. The mean total symptom score was 9.32 (range 3.00-14.00) and mean LES relaxation pressure before therapy was 56.29 mm Hg (range 8.00-107.80 mm Hg). Linear regression analysis showed a significant association between the total symptom score and LES relaxation pressure (p < 0.002, r = 0.290). Among the main symptoms, active and passive regurgitation showed significant correlation with LES relaxation pressure when compared to other individual symptoms using Pearson's correlation coefficient (p < 0.001 and 0.002, respectively). CONCLUSIONS: Our study showed that a clinical symptom score can be an appropriate predictor of the LES relaxation pressure in patients with idiopathic achalasia before therapy. Further studies are needed to evaluate similar correlations after therapeutic intervention. PMID- 12591042 TI - Function of the proximal stomach after partial versus complete laparoscopic fundoplication. AB - OBJECTIVES: After antireflux surgery, more than 30% of patients develop dyspeptic symptoms such as fullness and early satiety. We have previously shown that these symptoms are related to fundoplication-induced changes in proximal gastric motor and sensory function, especially impaired postprandial relaxation. We hypothesize that impaired fundus relaxation may be more pronounced after complete versus partial fundoplication. METHODS: Fasting and postprandial proximal gastric motor and sensory function were measured with an electronic barostat in patients after laparoscopic partial (n = 14) and complete (n = 14) fundoplication, in gastroesophageal reflux disease (GERD) patients (n = 12), and in healthy control subjects (n = 15). Gastric emptying and vagus nerve function tests were performed in all patients. RESULTS: Minimal distending pressure (MDP) and proximal gastric compliance were not significantly different among patients after antireflux surgery, GERD patients, and healthy controls. Maximal postprandial fundus relaxation was significantly (p < 0.01) reduced in patients after partial (267 +/ 32 ml) and complete (294 +/- 34 ml) fundoplication compared with GERD patients (448 +/- 30 ml) and healthy controls (409 +/- 25 ml). Sensations of fullness were not significantly different between patients with partial and complete fundoplication. There was a significant positive correlation between the postoperative duration and the degree of postprandial fundus relaxation (r = 0.67; p < 0.001). CONCLUSIONS: Both after complete and after partial fundoplication, proximal gastric motor function is affected, with impaired postprandial relaxation and increased sensation of fullness. These alterations are not related to the type of fundoplication but correlate significantly with the duration of the postoperative period. PMID- 12591043 TI - Which test to use to detect Helicobacter pylori infection in patients with low grade gastric mucosa-associated lymphoid tissue lymphoma? AB - Helicobacter pylori is involved in the pathogenesis of lymphoma of the gastric mucosa-associated lymphoid tissue (MALT). Because gastric MALT lymphoma is a rare disease, few studies comparing the accuracy of diagnostic tests in this group of patients have been carried out, and only a limited number of tests (essentially histological) were performed. The aim of our study was to compare the results of four different diagnostic methods used to detect H. pylori (histology, culture, polymerase chain reaction, and serology) in a prospective multicenter study. A patient was considered to be H. pylori positive if culture or histology was positive. During the period 1995-2000, a total of 90 patients with low-grade gastric MALT lymphoma were enrolled. Results for the four tests were available for 56 patients (62.2%). Among these patients, the four tests were concordant in 35 cases (62.5%), i.e., were positive in 19 cases (33.9%) and negative in 16 patients (17.8%). Histology (39/40 positive, 97.5%) and serology (38/40 positive, 95.0%) were the most sensitive tests. Polymerase chain reaction (PCR) and culture were positive in 52.5% and 50%, respectively. The cagA gene was detected in 47.4% of the strains. PMID- 12591044 TI - Accuracy of the stool antigen test for the diagnosis of childhood Helicobacter pylori infection: a multicenter Japanese study. AB - OBJECTIVE: The (13)C-urea breath test (UBT) has been accepted as a reliable noninvasive test for detecting Helicobacter pylori infection. Recently, another noninvasive test, a new enzyme immunoassay for H. pylori antigens in stool, has been widely investigated for its clinical usefulness. The purpose of this multicenter study was to evaluate the diagnostic accuracy of the stool antigen test in Japanese children. METHODS: A total of 264 children (148 male and 116 female; mean age 9.2 yr, range 2-17 yr) who underwent (13)C-UBT and the stool antigen test were studied. The diagnosis in these patients was gastritis (n = 49), gastric ulcer (n = 4), duodenal ulcer (n = 24), recurrent abdominal pain (n = 43), and other conditions (n = 144). The stool antigen test was performed using the HpSA ELISA (Premier Platinum HpSA, Meridian Diagnostics). According to manufacturer's instructions, an absorbance at 450/630 nm of <0.100, > or =0.120, and 0.100-0.119 was defined as negative, positive, and indeterminate, respectively. Based on the (13)C-UBT with a cutoff value of 3.5 per mil, the performance of HpSA was studied. In 21 patients who received eradication therapy, the HpSA was performed at baseline and at 1, 2, and 6 months after completion of therapy. Eradication of H. pylori was confirmed by (13)C-UBT at 2 or 3 months of follow-up. RESULTS: (13)C-UBT showed that 76 children were infected with H. pylori and 188 were not infected. In these same children, HpSA results were positive in 77 children, negative in 183, and indeterminate in four. The overall sensitivity, specificity, and accuracy of the test were 96.0% (95% CI = 88.6 99.2%), 96.8% (95% CI = 94.2-99.3%), and 96.5% (95% CI = 94.3-98.8%), respectively. There were no significant differences in these results among age groups of < or =5, 6-10, and > or =11 yr. Receiver operating characteristic curve analysis demonstrated that the best cutoff value of absorbance at 450/630 nm was 0.110. When a single cutoff value of 0.110 without indeterminate results was used, the sensitivity, specificity, and accuracy were 96.1% (95% CI = 90.8 99.7%), 96.3% (95% CI = 93.6-99.0%), and 96.2% (95% CI = 93.9-98.5%), respectively. In 19 patients in whom H. pylori was successfully eradicated, HpSA results were negative at 1 month of follow-up and remained negative through 6 months. CONCLUSIONS: The HpSA is an accurate test for the detection of H. pylori infection in all age groups of children. PMID- 12591045 TI - Maintenance oral pantoprazole therapy is effective for patients with Zollinger Ellison syndrome and idiopathic hypersecretion. AB - OBJECTIVE: Maintenance proton pump inhibitor (PPI) therapy is effective for gastric acid hypersecretory states, although data with pantoprazole are limited. The aim of this study was to evaluate the safety and efficacy of long term p.o. pantoprazole in individuals with hypersecretion. METHODS: All subjects had Zollinger-Ellison syndrome or idiopathic hypersecretion. Baseline acid output was measured in the presence of prior maintenance antisecretory therapy before pantoprazole exposure. The starting dose was 40 mg b.i.d. in most cases, and the dose was adjusted to document control within the first 2 wk of therapy. The maximal allowable dose was 240 mg daily. Acid output was measured on day 28 and then quarterly from month 3. The primary efficacy endpoint was documented control of acid secretion at 6 months, i.e., acid output in the last 1 h before the next dose of therapy of <10 mEq/h (<5 mEq/h in subjects with prior acid-reducing surgery). RESULTS: A total of 26 subjects had Zollinger-Ellison syndrome (six with multiple endocrine neoplasia syndrome type 1) and nine had idiopathic hypersecretion. Pre-enrollment therapy included omeprazole in 27 subjects and lansoprazole in eight, and 82.4% of subjects were controlled on their prior regimens. With upward dose titration, acid output was controlled in all subjects by day 10 and in all but two (6%) at the 6-month time point. Median acid secretion on therapy at 6 months was <2 mEq/h (mean 2.2 mEq/h; range 0-10.5 mEq/h) at a dose of 40 mg b.i.d. for 24 subjects, 80 mg b.i.d. for seven subjects, and 120 mg b.i.d. for two subjects. During the course of the study, five subjects required doses of 240 mg daily. Pantoprazole was generally well tolerated. No cases of anterior optic ischemic neuropathy occurred. Five subjects died during follow-up, all because of events unrelated to the study drug. CONCLUSIONS: Maintenance p.o. pantoprazole therapy at a dose of 80-240 mg/day in divided doses was both effective and generally well tolerated for patients with Zollinger-Ellison syndrome and idiopathic hypersecretion. PMID- 12591046 TI - Safety of ERCP during pregnancy. AB - OBJECTIVES: There are few data in the literature regarding the indications, therapy, and safety of endoscopic management of pancreatico-biliary disorders during pregnancy. We report the largest single center experience with ERCP in pregnancy. METHODS: We reviewed 15 patients that underwent ERCP during pregnancy. In all patients, the pelvis was lead-shielded and the fetus was monitored by an obstetrician. Fluoroscopy was minimized and hard copy radiographs taken only when essential. RESULTS: The mean patient age was 28.9 yr (15-36 yr). The mean duration of gestation was 25 wk (12-33 wk); one patient was in the first, five in the second, and nine in the third trimester. The indications were gallstone pancreatitis (n = 6), choledocholithiasis on ultrasound (n = 5), elevated liver enzymes and a dilated bile duct on ultrasound (n = 2), abdominal pain and gallstones (n = 1), and chronic pancreatitis (n = 1). ERCP findings were bile duct stones (n = 6), patulous papilla (n = 1), bile duct debris (n = 1), normal bile duct and gallstones or gallbladder sludge (n = 3), dilated bile duct and gallstones (n = 1), normal bile duct and no gallstones (n = 2), and chronic pancreatitis (n = 1). Six patients underwent sphincterotomies and one a biliary stent insertion. One sphincterotomy was complicated by mild pancreatitis. All infants delivered to date have had Apgar-scores >8, and continuing pregnancies are uneventful. Mean fluorosocopy time was 3.2 min (SD +/- 1.8). An estimated fetal radiation exposure was 310 mrad (SD +/- 164) which is substantially below the accepted teratogenic dose. CONCLUSIONS: ERCP in pregnancy seems to be safe for both mother and fetus; however, it should be restricted to therapeutic indications with additional intraprocedure safety measures. PMID- 12591047 TI - Electroacupuncture analgesia for colonoscopy. a prospective, randomized, placebo controlled study. AB - OBJECTIVE: A study was undertaken to compare the effectiveness of acupuncture in reducing patient's discomfort and anxiety during colonoscopy with a standard sedation protocol. METHODS: Thirty patients scheduled to undergo colonoscopy were randomly assigned to receive acupuncture, sham, or no acupuncture. A verbal rating scale was used to measure patient's pain when the endoscope reached four scheduled positions. Midazolam was administered at baseline and again anytime patients complained of "severe" pain. The amount of midazolam administered was recorded. A verbal rating scale was used to assess patient's satisfaction with the level of sedation achieved. RESULTS: Pain level was lower, although not significantly, in the acupuncture group. Midazolam boluses were required in three patients with acupuncture (30%), eight in the sham group (80%), and nine in the control group (90%) (p = 0.01). Six patients in the acupuncture group (60%) reported optimum acceptance of colonoscopy compared with only one in the sham group (10%) and none in the control group (0%) (p = 0.016). Satisfaction scores, assessed 24-72 h after colonoscopy, were extremely high (median score 90 of 100) in the three groups. CONCLUSIONS: Acupuncture may decrease the demand for sedative drugs during colonoscopy by reducing discomfort and anxiety of the patient and the well-known adverse effects of pharmacologic sedation. PMID- 12591048 TI - Timing of colonoscopy: impact on length of hospital stay in patients with acute lower intestinal bleeding. AB - OBJECTIVE: Previous studies suggest that urgent colonoscopic evaluation of massive lower intestinal bleeding (LIB) can reduce hospital length of stay (LOS). We sought to determine if time to colonoscopy impacts hospital LOS in patients admitted with all sources and severities of acute LIB. METHODS: A total of 252 consecutive patients admitted to a tertiary care hospital with acute LIB were identified. Cox proportional hazards regression was used to determine independent predictors of hospital LOS. Time from admission to colonoscopy was analyzed as a time-varying covariate. RESULTS: A total of 144 patients (57%) underwent an inpatient colonoscopy: 14 were done in <12 h, 55 in 12-24 h, 46 in 24-48 h, and 29 in >48 h. After controlling for the other independent correlates, earlier colonoscopy was significantly associated with a shorter hospital LOS (hazards ratio = 2.02, 95% CI = 1.5-2.6, p < 0.0001). The absence of visible blood or active bleeding at the time of colonoscopy was also independently related to a shorter hospital LOS (hazards ratio = 1.5, 95% = CI 1.1-2.0, p = 0.01). CONCLUSIONS: Time to colonoscopy is an independent predictor of hospital LOS. In a wide spectrum of patients with LIB, this reduction in hospital LOS seems to be primarily related to improved diagnostic yield rather than therapeutic interventions. PMID- 12591049 TI - Salivary gland involvement in chronic pancreatitis of various etiologies. AB - OBJECTIVE: Both the pancreas and salivary glands show many histological and functional similarities. Recently, autoimmune pathogenesis has been postulated in some chronic pancreatitis cases. To examine whether a cell-mediated phenomenon involving the pancreas has a secondary effect on the salivary glands, we assessed the frequency of salivary gland dysfunction in patients with chronic pancreatitis of various etiologies. METHODS: Function of the salivary glands was examined by sialochemistry and salivary gland scintigraphy in patients with chronic pancreatitis (n = 33), Sjogren's syndrome (n = 45), and controls (n = 28). Etiologies of chronic pancreatitis were alcoholic (19 cases), idiopathic (seven cases), and autoimmune (seven cases). RESULTS: Concentrations of Na+, amylase, and beta2-microglobulin in saliva were investigated. In submandibular and parotid gland scintigraphy, time-activity curves were generated, and the ratios of peak count density and washout were calculated. Concentrations of Na+ in saliva of patients with idiopathic chronic pancreatitis and of beta2-microglobulin in saliva of patients with idiopathic and autoimmune chronic pancreatitis were significantly elevated than those of the control group. In submandibular and parotid gland scintigraphy, the peak count density ratio of patients with all chronic pancreatitis and washout ratio of patients with alcoholic and idiopathic chronic pancreatitis were significantly lower than those of the control group. CONCLUSIONS: Salivary gland function was frequently impaired in the course of chronic pancreatitis of various etiologies. Salivary gland dysfunction might be the result of a common pathophysiological effect of alcohol in patients with alcoholic chronic pancreatitis and the aggressive immune mechanism against the pancreatic and the salivary ducts in patients with autoimmune and idiopathic chronic pancreatitis. PMID- 12591050 TI - Is irritable bowel syndrome more common in patients presenting with bacterial gastroenteritis? A community-based, case-control study. AB - OBJECTIVE: Irritable bowel syndrome (IBS) has been reported to follow infectious diarrhea. Food-borne infections affect 76 million people in the United States and 9.4 million in England per year; of these, only a small percentage of patients see their doctor, and even fewer will have stool culture confirmation. We hypothesized that patients who present to their doctor with gastroenteritis and have positive stool samples may be different from the normal population with regard to their pre-existing bowel symptoms. Our aim was to determine if patients with bacterial gastroenteritis were more likely to have prior IBS, functional dyspepsia, or functional diarrhea, compared with a control population. METHODS: Between January, 2000 and January, 2001, subjects with stool positive bacterial gastroenteritis and control subjects from the same primary care practice were invited to participate. The main outcome measure was the presence of IBS, functional dyspepsia, or functional diarrhea diagnosed using self-report Rome II modular questionnaires. RESULTS: A total of 217 people with recent bacterial gastroenteritis and 265 community controls consented to participate in the study. Of these, 89/217 cases and 46/265 controls had one of the functional GI disorders (OR = 3.3; 95% CI = 2.17-5.00). IBS was present in 67 cases (31%) and 26 controls (10%) (OR = 4.1; 95% CI = 2.49-6.72). There was no statistically significant difference in the presence of prior functional dyspepsia or functional diarrhea. CONCLUSIONS: IBS is more frequent before diagnosis in people with bacterial gastroenteritis presenting to their primary care physician than in community controls. Studies that examine the rate of IBS after bacterial gastroenteritis need to carefully exclude people with prior IBS in a systematic way. PMID- 12591051 TI - Magnetic resonance imaging of the effects of infliximab on perianal fistulizing Crohn's disease. AB - OBJECTIVES: Although the clinical efficacy of infliximab as measured by closure of fistulas in Crohn's disease has been demonstrated, its influence on the inflammatory changes in the fistula tracks is less clear. The aim of the present study was to assess the behavior of perianal fistulas before and after infliximab treatment. METHODS: Magnetic resonance imaging (MRI) and clinical evaluation were performed in a total of 18 patients before and after treatment with infliximab. An MRI-based score of perianal Crohn's disease severity was developed using both criteria of local extension of fistulas (complexity, supralavetoric extension, relation to the sphincters and of active inflammation (T2 hyperintensity, presence of cavities/abscesses, and rectal wall involvement). RESULTS: The MRI score was reliable in assessing the fistula tracks, with a good interobserver concordance (p < 0.001). Fistula tracks with signs of active inflammation were found in all 18 patients at baseline and collections in seven. After short-term infliximab treatment, active tracks persisted in eight of 11 patients who had clinically responded to infliximab. After long-term (46 wk) infliximab therapy, MRI signs of active track inflammation had resolved in three of six patients. CONCLUSIONS: We have developed an MRI-based score of perianal Crohn's disease severity to assess the anatomical evolution of Crohn's fistulas. Our study demonstrates that despite closure of draining external orifices after infliximab therapy, fistula tracks persist with varying degrees of residual inflammation, which may cause recurrent fistulas and pelvic abscesses. Whether complete fistula fibrosis occurs over time with repeated infliximab infusions needs further study. PMID- 12591052 TI - Collagenous and lymphocytic colitis. evaluation of clinical and histological features, response to treatment, and long-term follow-up. AB - OBJECTIVE: Data on collagenous colitis (CC) and lymphocytic colitis (LC) have been based on retrospective studies of registries of patients from multiple hospitals. Such studies may induce a selection of patients with severe forms of the disease, and conclusions about the clinical spectrum of the disease and treatment efficacy are difficult to be drawn. The aim of this study was to compare the clinical features, response to treatment, and long-term follow-up of CC and LC in a large group of patients prospectively diagnosed in a single center. METHODS: A specific program was undertaken to prospectively diagnose all patients with microscopic colitis from those referred for a full colonoscopy because of recurrent or chronic diarrhea. Detailed clinical and histological features, response to treatment, and long-term follow-up were compared in patients with confirmed CC and LC. RESULTS: Thirty-seven patients with CC and 44 with LC were included. Patients with CC were significantly younger and had a significantly longer duration of diarrhea before diagnosis than those with LC. Otherwise, clinical presentation was similar. Drug-induced disease was suspected for ticlopidine, flutamide, gold salts, and bentazepam in LC. Complete resolution of diarrhea was achieved in all patients, spontaneously occurring in nearly 20% of them. Response to salicylates (mainly, mesalazine) was significantly better in LC than in CC (86% vs 42%, p = 0.005). Cholestyramine was highly effective in patients of both groups with concomitant bile acid malabsorption. Patients with CC required prednisone more often than those with LC (30% vs 4.5%, p = 0.005). Both prednisone and budesonide controlled ileal release were highly effective in patients with CC (82% and 89% efficacy). After cessation of diarrhea, 25% of patients with LC and 30% of those with CC relapsed after a mean follow-up of around 3 yr. CONCLUSIONS: CC and LC share a similar clinical picture and have a benign course with long-term cessation of diarrhea in more than 70% of patients. Mesalazine and budesonide seem to be good options as first-line treatment in LC and CC, respectively. Cholestyramine may be a good alternative in patients with concomitant bile acid malabsorption. PMID- 12591053 TI - Antioxidant vitamin supplementation in Crohn's disease decreases oxidative stress. a randomized controlled trial. AB - OBJECTIVE: We showed previously that patients with Crohn's disease (CD) had increased oxidative stress and lower antioxidant vitamins compared with healthy controls. This is despite inactive or mildly active disease and maintenance therapy. The aim of this study was to evaluate in these patients the effects of antioxidant vitamin supplementation on oxidative stress. METHODS: This is a randomized controlled trial where stable but oxidatively stressed CD subjects (n = 57) were supplemented with vitamins E (800 IU) and C (1000 mg) or their placebo for 4 wk. Oxidative stress measured by breath pentane and ethane output, plasma lipid peroxides, and F2-isoprostane was assessed at baseline and at 4 wk. Disease activity was also monitored by measuring CD activity index and plasma orosomucoid. RESULTS: During supplementation, plasma vitamin C and alpha tocopherol increased and all indices of oxidative stress decreased significantly. Disease activity remained stable. CONCLUSIONS: In this population, vitamin E and C supplementation resulted in a significant reduction in oxidative stress. This suggests that patients with inactive or mildly active CD can be oxidatively stressed and have increased requirement in antioxidant vitamins. PMID- 12591054 TI - Effectiveness and appropriateness of empiric metronidazole for Clostridium difficile-associated diarrhea. AB - OBJECTIVE: Although Clostridium difficile is the most common infectious etiology of nosocomial diarrhea, noninfectious causes are far more common. Empiric initiation of therapy for all patients is of unknown value. The aim of this study was to determine benefits of empiric metronidazole for Clostridium difficile associated diarrhea (CDAD). METHODS: We conducted a 4-month prospective surveillance of all patients in two community teaching hospitals receiving metronidazole for empiric treatment of presumptive CDAD. A database including antibiotic usage, fever, white blood cell count, feeding formula usage, comorbidity, and response to therapy was maintained. RESULTS: Seventy-one patients on the medical (50), surgical (18), obstetric (two), and trauma (one) service were identified. Sixty-two had nosocomial diarrhea; nine had diarrhea on admission. Seventy (97%) received antibiotics; one (3%) was on nelfinavir only. Eighteen (25%) were subsequently proven to have CDAD; two (3%) had laxative induced diarrhea; two (3%) had diarrhea secondary to a medication (colchicine [one] and nelfinavir [one]); one (1%) had diarrhea caused by bowel preparation for colonoscopy. The remaining 49 (68%) did not have a clearly established diarrhea etiology. (Four did not undergo stool examination.) Statistical analysis (chi(2) test) demonstrated a significant decrease in symptoms for metronidazole treated patients with CDAD versus those with a different diagnosis (p = 0.05). Not surprisingly, multivariate regression analysis identified a strong correlation of diagnosing CDAD with age >60 yr, antibiotics exposure, fever, elevated white blood cell count, and resolution of symptoms with specific metronidazole treatment. CDAD was definitively diagnosed in 25% of our hospitalized patients with diarrhea, consistent with published data. Although some cases might have been missed, most patients did not have CDAD and received no benefit (and were potentially harmed) by empiric metronidazole. There was no way a priori to distinguish CDAD from non-CDAD. CONCLUSIONS: In the absence of clear guidelines, empiric metronidazole should be reserved for strongly presumptive CDAD patients (older patients with comorbid conditions receiving broad-spectrum antibiotics associated with CDAD) who cannot hemodynamically or otherwise tolerate diarrhea. Used judiciously, empiric therapy may more rapidly resolve symptoms, and could conceivably prevent/abate severe complications and nosocomial spread. PMID- 12591055 TI - Association of MICA-A5.1 allele with susceptibility to celiac disease in a family study. AB - OBJECTIVE: The aim of this study was to analyze the role of the major histocompatibility complex class I chain-related gene A (MICA) transmembrane polymorphism in celiac disease (CD) susceptibility. METHODS: Sixty-one celiac Spanish families were genotyped for MICA transmembrane polymorphism by a polymerase chain reaction method combined with fluorescent technology. A transmission disequilibrium test was performed to investigate the preferential transmission of MICA alleles to the affected offspring. RESULTS: The MICA A5.1 allele was shown to be significantly transmitted to the affected siblings. This association was independent of the CD-predisposing DQ2 haplotype. Additionally, we classified our celiac families into typical and atypical groups as we found a significant association with MICA A5.1 in typical celiac families. There was also an association tendency with atypical families. CONCLUSIONS: Our data suggest that the MICA A5.1 allele is associated with CD development independently of DQ2 extended haplotype and clinical forms of CD. PMID- 12591056 TI - Tobacco consumption and disease duration are associated with fistulizing and stricturing behaviors in the first 8 years of Crohn's disease. AB - OBJECTIVES: Crohn's disease (CD) can be classified by site of involvement and by clinical behavior. Claims for stability of behavior are based on patients who have had surgery, usually within 8 yr of diagnosis. Disease during this period may begin as inflammatory and may evolve into stricturing or fistulizing behavior. Our aim was to determine the influence of genetic and environmental factors on the prevalence of inflammatory behavior among patients who have had CD for <8 yr. METHODS: Disease type (inflammatory, stricturing, or fistulizing), site, and duration were determined in 311 consecutive patients with CD. The analysis was then restricted to those with a disease duration of <8 yr, and influences on the prevalence of inflammatory type disease were compared with those of the complicated type (fistulizing and stricturing), including disease site, family history, Jewish ethnicity, and pack-years of cigarette smoking after diagnosis were determined through univariate and multivariate analyses. RESULTS: The prevalence of inflammatory type disease was 0.63 and 0.3 (p < 0.0001) in patients with a disease duration of <8 yr and > or =8 yr, respectively. Multivariate analysis revealed a 91% decrease in the odds of inflammatory disease among those with > or =1 pack-year of smoking after diagnosis among individuals with disease <8 yr. This was not influenced by disease location, family history of inflammatory bowel disease (IBD), or Jewish ethnicity. CONCLUSIONS: The prevalence of inflammatory CD decreased with time. Tobacco consumption was associated with this decline during the early phase of disease, suggesting that tobacco may influence the progression of inflammatory to stricturing or fistulizing type disease. PMID- 12591057 TI - A pilot study on the use of the humanized anti-interleukin-2 receptor antibody daclizumab in active ulcerative colitis. AB - OBJECTIVES: Medical therapy of refractory ulcerative colitis (UC) is associated with long-term side effects of cyclosporine and steroids. Because cyclosporine acts by inhibiting interleukin-2 (IL-2) production, we studied the efficacy and safety of humanized anti-IL2 receptor (CD25) antibodies daclizumab for refractory UC in an open label pilot study. METHODS: Ten patients with chronically active UC received daclizumab, 1 mg/kg i.v. twice with a 4-wk interval. Clinical, endoscopic, and histological evaluation was scored at regular intervals. CD25 immunohistochemistry was followed in mucosal biopsies. The primary study endpoint was clinical improvement at wk 8. RESULTS: Nine of 10 patients completed the study. The median clinical activity score decreased from a median of 8 (95% CI = 7.2-9.2) at baseline to 3.5 (95% CI = 1.4-4.9) at wk 8 (p < 0.005). Endoscopic scores were significantly decreased at wk 8 (wk 0: 8, 95% CI = 6.3-8.5; wk 8: 5.0, 95% CI = 2.6-6.3; p < 0.01). Mucosal biopsies showed a significant decrease in CD25+ cells, and there was a trend toward lower histology scores at wk 8. Quality of life as assessed by the Inflammatory Bowel Disease Questionnaire increased after therapy (baseline: 131, 95% CI = 119-178; wk 8: 169; 95% CI = 124 216, p < 0.05). Nausea was most frequently reported as an adverse event, but always in patients that were concomitantly started on azathioprine. CONCLUSIONS: The anti-IL-2R antibody daclizumab was safe and well tolerated in acute UC. Patients experienced clinical benefit along with signs of endoscopic improvement, but further controlled trials are needed to determine the therapeutic benefit of this compound. PMID- 12591058 TI - Prevalence of celiac disease among relatives of sib pairs with celiac disease in U.S. families. AB - OBJECTIVE: Celiac disease is a familial malabsorptive disorder with an estimated prevalence in first-degree relatives of 10-12%. The prevalence for first-degree and more distant relatives has not been determined in families where there are two affected first-degree relatives. The aim of our investigation was to estimate the prevalence and relative risk for celiac disease in relatives of two siblings diagnosed with celiac disease. METHODS: We ascertained sib pairs with celiac disease, and then identified all living first-degree relatives and available second-degree relatives to minimize ascertainment bias. We measured IgA endomysial antibodies, a highly specific and sensitive assay for celiac disease, in all subjects without a confirmed biopsy diagnosis. For those individuals with positive serologic tests, IgA tissue transglutaminase antibody tests and human leukocyte antigen DQA1 and DQB1 genotyping were performed for additional confirmation. Individuals with positive biopsy and/or serology were considered affected. We calculated the relative risk of being affected with celiac disease using the lambda(R) statistic. RESULTS: The prevalence of celiac disease in relatives of affected sib pairs was as follows: 21.3% (13/61) of siblings (lambda(S) = 53); 14.7% (10/68) of offspring (lambda(O) = 37); 17.2% (28/163) of first-degree relatives; 19.5% (16/82) of second-degree relatives; and 17.8% (52/292) of all relatives (lambda(R) = 44.5). CONCLUSIONS: In these families, we identified a sibling risk approximately double that found in previous reports, as well as significant risk for more distant relatives, probably because of sharing of a common gene. In families where at least two siblings have been diagnosed with celiac disease, relatives are at high risk for celiac disease. Screening should be considered for all family members. PMID- 12591059 TI - Analysis of the structure and strength of bones in celiac disease patients. AB - OBJECTIVE: The aim of this study was to gain insight into the pathogenesis of bone mass loss and weakening affecting patients with celiac disease, according to the new concepts of bone structure/strength and muscle/bone interrelationships. METHODS: We studied serum variables and tomographic indicators of bone structure and strength and regional muscle masses in a series of patients at diagnosis and after 1 yr on a gluten-free diet and in gender- and age-matched controls. RESULTS: At diagnosis, serum levels of calcium and vitamin D were low, and indicators of parathyroid hormone activity and bone formation and resorption were increased. All these parameters were normalized by treatment. Peripheral quantitative CT scans of the distal radius revealed that cortical bone was generally more affected than trabecular bone. The cross-sectional area (CSA) and the volumetric mineral content and density of cortical bone (indicators of cortical tissue mass and mechanical quality), and moments of inertia (CSMI, indicator of the bone architectural design) were in the lower end of normal range in men, and below that in 50% of women. In men, the CSMI/CSA ratio (indicator of the architectural efficiency of distribution of the available cortical tissue) was lower than expected and remained unchanged after treatment. In women, the baseline ratio was normal, but both the ratio and the CSMI were low at diagnosis and normalized after treatment. Both baseline values and the treatment-induced changes of cortical and trabecular bone in the radius and the axis (L3) correlated inversely with serum parathyroid hormone levels. Baseline values or changes in the mineral content of the vertebral bone correlated with the CSA of psoas and spine-extensor muscles. Multiple regression analyses showed that metabolic and mechanical parameters were independent determinants of different aspects of the vertebral bone weakening. CONCLUSIONS: Our results show that bone weakening in celiac disease might result from both 1) a metabolic disturbances of bone remodeling affecting trabecular and cortical bone masses and the mechanical quality of the bone material, and 2) a reduction of muscle strength impairing the modeling-dependent optimization of bone architectural design and mass of cortical bone. Dietary treatment seems to correct almost exclusively the metabolically induced disturbances, which were predominant in women. PMID- 12591060 TI - Symptom clustering in subjects with and without diabetes mellitus: a population based study of 15,000 Australian adults. AB - OBJECTIVES: GI symptoms form distinct symptom clusters in community samples when factor and cluster analysis is applied. However, this has not been studied in diabetic populations, despite clear evidence that GI complaints are common in patients with diabetes mellitus (DM). This study aimed to describe clustering of GI symptoms among individuals with and without diabetes mellitus, and to describe associations of symptom clustering in diabetes mellitus, with self-reported glucose control and treatment. METHODS: A large population survey (n = 15,000) was used to identify a cohort with diabetes mellitus. Items assessing therapy and quality of glycemic control were included, as were those assessing 16 common GI symptoms. Latent GI symptom factors were extracted by factor analysis and used in a k-means cluster analysis. The latter serves to group individuals according to commonalities in symptom profiles. The association of cluster group membership to glycemic control and diabetic treatment was described by logistic regression. RESULTS: Factor analysis identified four latent symptom factors, which accounted for 69.3% of the total variance. These were labeled Upper GI/Dysmotility, Diarrhea, Constipation, and Vomiting/Nausea. The k-means analysis produced a five cluster solution, which included a "health" group and four "diseased" groups, each identified by a predominant symptom: Upper GI/Dysmotility symptoms, Nausea/Vomiting, Diarrhea, and Constipation. After adjustment for age and gender, poor glycemic control predicted membership in all disease clusters, when compared separately with the health group. Oral hypoglycemic drugs predicted membership in the Nausea/Vomiting cluster (OR = 5.13) when used alone, and membership in the Nausea/Vomiting (OR = 10.12) and Upper GI/Dysmotility cluster (OR = 10.12) when used in combination with insulin. CONCLUSION: Diabetes can be grouped according to common GI symptoms. Glycemic control and treatment for DM predict membership of symptom clusters. PMID- 12591061 TI - Magnetic resonance imaging of anatomic and dynamic defects of the pelvic floor in defecatory disorders. AB - OBJECTIVE: Endoanal ultrasound identifies anal sphincter anatomy, and evacuation proctography visualizes pelvic floor motion during simulated defecation. These complementary techniques can evaluate obstructed defecation and fecal incontinence. Our aim was to develop a single, nonionizing, minimally invasive modality to image global pelvic floor anatomy and motion. METHODS: We studied six patients with fecal incontinence and seven patients with obstructed defecation. The anal sphincters were imaged with an endoanal magnetic resonance imaging (MRI) coil and endoanal ultrasound (five patients). MR fluoroscopy acquired images every 1.4-2 s, using a modified real-time, T2-weighted, single-shot, fast-spin echo sequence, recording motion as patients squeezed pelvic floor muscles and expelled ultrasound gel; no contrast was added to other pelvic organs. Six patients also had scintigraphic defecography. RESULTS: Endoanal ultrasound and MRI were comparable for imaging defects of the internal and external sphincters. Only MRI revealed puborectalis and/or external sphincter atrophy; four of these patients had fecal incontinence. MR fluoroscopy recorded pelvic floor contraction during squeeze and recorded relaxation during simulated defecation. Corresponding comparisons for angle change and perineal descent during defecation were not significant; only MRI, but not scintigraphy, identified excessive perineal descent in two patients. CONCLUSIONS: Pelvic MRI is a promising single, comprehensive, nonradioactive modality to measure structural and functional pelvic floor disturbances in defecatory disorders. This method may provide insights into mechanisms of normal and disordered pelvic floor function in health and disease. PMID- 12591062 TI - Normalization of lactulose breath testing correlates with symptom improvement in irritable bowel syndrome. a double-blind, randomized, placebo-controlled study. AB - OBJECTIVE: We have recently found an association between abnormal lactulose breath test (LBT) findings and irritable bowel syndrome (IBS). The current study was designed to test the effect of antibiotic treatment for IBS in a double-blind fashion. METHODS: Consecutive IBS subjects underwent an LBT with the results blinded. All subjects were subsequently randomized into two treatment groups (neomycin or placebo). The prevalence of abnormal LBT was compared with a gender matched control group. Seven days after completion of treatment, subjects returned for repeat LBT. A symptom questionnaire was administered on both days. RESULTS: After exclusion criteria were met, 111 IBS subjects (55 neomycin, 56 placebo) entered the study, with 84% having an abnormal LBT, compared with 20% in healthy controls (p < 0.01). In an intention-to-treat analysis of all 111 subjects, neomycin resulted in a 35.0% improvement in a composite score, compared with 11.4% for placebo (p < 0.05). Additionally, patients reported a percent bowel normalization of 35.3% after neomycin, compared with 13.9% for placebo (p < 0.001). There was a graded response to treatment, such that the best outcome was observed if neomycin was successful in normalizing the LBT (75% improvement) (one way ANOVA, p < 0.0001). LBT gas production was associated with IBS subgroup, such that methane excretion was 100% associated with constipation-predominant IBS. Methane excretors had a mean constipation severity of 4.1, compared with 2.3 in all other subjects (p < 0.001). CONCLUSIONS: An abnormal LBT is common in subjects with IBS. Normalization of LBT with neomycin leads to a significant reduction in IBS symptoms. The type of gas seen on LBT is also associated with IBS subgroup. PMID- 12591063 TI - Symptoms across the menstrual cycle in women with irritable bowel syndrome. AB - OBJECTIVE: The purpose of this study was to describe the patterns of GI, somatic, and psychological symptoms across the menstrual cycle in women with irritable bowel syndrome, and to determine whether symptoms differed by oral contraceptive use or predominant bowel pattern. METHODS: A daily diary was used to assess symptoms across one menstrual cycle. Repeated-measures analysis of covariance, controlling for age and body mass index, was used to compare patterns of symptoms across the menstrual cycle by oral contraceptive use and predominant bowel pattern (diarrhea, constipation, alternating). Data from control women are presented for comparison. RESULTS: For somatic and psychological as well as GI symptoms, women with irritable bowel syndrome had higher symptom severity than did controls. Women with irritable bowel syndrome using oral contraceptives had lower cognitive, anxiety, and depression symptoms (p < 0.05, but not significant after multiple comparison adjustment), but no differences were seen for most symptoms of irritable bowel syndrome. All symptoms except diarrhea were highest in the alternating group and lowest in the diarrhea group, with the constipation group either intermediate or close to the alternating group. This pattern was significant after multiple comparisons adjustment for GI symptoms, and trending toward significance (p < 0.05, but not significant after multiple comparison adjustment) for menstrual, sleep, and cognitive symptoms. The strongest menstrual cycle effect was seen in somatic and menstrual symptoms. The pattern of symptoms over the menstrual cycle did not differ by predominant bowel pattern or by oral contraceptive use. CONCLUSIONS: Many of the symptoms examined differed by predominant bowel pattern and menstrual cycle phase, not just the GI symptoms. The menstrual cycle variation was similar regardless of oral contraceptive use or predominant bowel pattern. PMID- 12591064 TI - Characterization and clinical impact of antinuclear antibodies in primary biliary cirrhosis. AB - OBJECTIVES: The clinical impact of antinuclear antibodies in primary biliary cirrhosis is uncertain. We analyzed in detail the antinuclear antibodies reactivity of primary biliary cirrhosis patients and correlated the fine specificities observed with clinical, biochemical, and immunologic parameters. METHODS: A total of 96 consecutive primary biliary cirrhosis patients and 283 pathologic controls were studied. To dissect the fine antinuclear antibodies specificities we used different techniques, such as indirect immunofluorescence on cryostat tissue sections and cell culture (HEp-2 cells), counterimmunoelectrophoresis with thymus and spleen extracts, ELISA assays with recombinant Sp100 and purified gp210 and Lamin B receptor, and immunoblot with several recombinant nuclear and cytoplasmic antigens. RESULTS: Antinuclear antibodies were detected in 53% of patients, with the following hierarchy of specificities: 27% anti-Sp100, 16% "multiple nuclear dots," 16% anti-gp210, 16% anti-centromere, 7% XR1, 6% anti-lamin B receptor, 5% anti-SS-A/Ro, 5% anti ribonucleoprotein, 4% XR2, 2% anti-SS-B/La, 2% perinuclear antineutrophil cytoplasmic antibodies, and 1% anti-double-stranded deoxyribonucleic acid. Several patients showed multiple specificities. The "multiple nuclear dots" pattern was detected more often in antimitochondrial antibodies negative patients. In particular, primary biliary cirrhosis specific antinuclear antibodies (anti-Sp100, anti-gp210, and anti-lamin B receptor) were detected in nine of 13 antimitochondrial negative primary biliary cirrhosis cases. Anti-gp210 was more frequent in patients with more pronounced cholestasis and more impaired liver function. CONCLUSIONS: Antinuclear antibodies reactivities are present in more than half of primary biliary cirrhosis patients and target diverse autoantigens located in distinct subnuclear structures. Anti-gp210 identifies a subgroup of primary biliary cirrhosis patients with more serious liver disease. Positivity for anti-Sp100, anti-gp210, and anti-lamin B receptor, either alone or in combination, may act as a serologic marker of antimitochondrial antibodies negative primary biliary cirrhosis. PMID- 12591065 TI - Association between hepatitis C, diabetes mellitus, and race. a case-control study. AB - OBJECTIVE: A higher prevalence of type II diabetes mellitus (DM) has been reported in patients with hepatitis C virus (HCV) infection. However, in most of these studies, the control population was not matched for body mass index, race, and severity of liver disease, known risk factors for the development of type II DM. The aim of this study was to determine the prevalence of type II DM in patients with HCV cirrhosis compared with a control population matched for age, sex, body mass index, and severity of liver disease. METHODS: We conducted a case control study in a University Hospital setting. We compared 97 cirrhotic patients with HCV (cases) with 194 HCV-negative patients with cirrhosis from other causes (controls). We sought to determine the prevalence of pre- and post-transplant type II DM in cases and controls. RESULTS: The age, sex, and severity of liver disease were similar in both groups, but there were more blacks in the HCV group (24 of 97, 25%) compared with controls (16 of 194, 8%). The prevalence of pretransplant DM was higher in the HCV group (19.6%) compared with controls (11.5%) (p = 0.06, OR = 1.9, 95% CI = 0.9-3.8). Blacks with HCV had a significantly higher prevalence of pretransplant DM (33.3%) compared with whites with HCV (13.2%) (p = 0.03) and black controls (6.3%) (p = 0.05). Among whites, the prevalence of DM was similar in the HCV group (13.2%) and controls (11.9%). Logistic regression showed that age was the only independent predictor for pretransplant DM (odds ratio = 1.06, 95% CI = 1.01-1.11, p = 0.01). New onset DM was similar in the HCV group (16.7%) and controls (10.1%, p = ns). The new onset of DM was similar in blacks with HCV (31.3%) and black controls (20.0%). However, by logistic regression, black race was an independent predictor for the development of new onset DM (odds ratio = 3.4, 95% CI = 1.2-9.8, p = 0.02). CONCLUSIONS: Our study shows that the prevalence of type II DM is higher in patients with HCV cirrhosis compared with a control group of patients with cirrhosis from other causes, and this was because of a higher prevalence of DM in blacks with HCV infection. PMID- 12591066 TI - Mutations of the HFE gene in patients with hepatocellular carcinoma. AB - OBJECTIVE: Hepatocellular carcinoma (HCC) is a late consequence of severe liver disease. Patients with genetic hemochromatosis may be at risk for HCC, but limited information is available on the relationship of HCC and heterozygosity for the HFE gene mutations. METHODS: HFE mutations (C282Y and H63D) were assessed in 162 consecutive patients (131 men/31 women) with HCC. A total of 159 patients had cirrhosis. The most common etiologies of cirrhosis were chronic viral hepatitis (hepatitis C 39%, hepatitis B 9%) and alcoholic liver disease (36%). RESULTS: Five patients were C282Y homozygotes, four C282Y/H63D compound heterozygotes, and three H63D homozygotes. The C282Y and H63D allele frequencies in HCC were 8.3 (95% confidence limit = 5.3-11.3) and 11.1 (7.8-14.6), respectively, and not different from previously published data in healthy subjects or patients with chronic hepatitis C in Austria. Furthermore, there was no difference in the age at diagnosis in patients with or without HFE gene mutations. C282Y homozygotes had a 19-fold increased risk to develop HCC. In contrast, all other HFE allele constellations were not associated with such a risk. CONCLUSIONS: Except for C282Y homozygotes, HFE gene mutations do not increase the risk to develop HCC in patients with cirrhosis. PMID- 12591067 TI - Viral hepatitis-related acute liver failure. AB - OBJECTIVES: Viral hepatitis has previously been the major cause of acute liver failure (ALF) in the United States. We aimed to determine the incidence of viral hepatitis-related ALF and to compare the outcome and clinical and biochemical variables in patients with hepatitis A and B. METHODS: A total of 354 patients with ALF from multiple centers were screened for possible acute viral etiology. RESULTS: Forty-three patients (12.1% of all ALF cases) had acute viral hepatitis: hepatitis A (n = 16), hepatitis B (n = 26), and herpes simplex virus infection (n = 1). There was no difference between groups with regard to age, gender, body mass index, admission or peak coma grade, symptom duration, admission mean arterial pressure, temperature, or biochemical liver tests, creatinine, arterial pH, or rate of infections. Platelet count was significantly higher in hepatitis A patients than in hepatitis B patients. The transplantation-free (spontaneous) survival rate was significantly higher for hepatitis A patients (69%) than for hepatitis B patients (19%, p = 0.007), whereas the liver transplantation rate was higher in hepatitis B patients (62%) than in hepatitis A patients (19%, p = 0.017). Spontaneous survivors had significantly higher mean arterial pressure, higher platelet count, and lower AST/ALT ratio than patients who did not survive spontaneously. CONCLUSIONS: Viral hepatitis now comprises only one-eighth of all ALF cases in the United States. The marked difference in spontaneous survival between hepatitis A and B cannot be explained by the severity of hepatic dysfunction on admission but may rather be an inherent feature of the infections or a bias toward transplanting patients with hepatitis B. PMID- 12591068 TI - Risk of esophageal variceal bleeding based on endoscopic ultrasound evaluation of the sum of esophageal variceal cross-sectional surface area. AB - OBJECTIVE: The aim of this study was to evaluate the risk of future variceal bleeding, based on the endoscopic ultrasound measurement of the sum of the cross sectional surface area (CSA) of all of the esophageal varices in the distal esophagus. METHODS: Twenty-eight patients with portal hypertension and esophageal varices, but no prior history of variceal bleeding, were evaluated using endoscopic ultrasound (20-MHz ultrasound probe, Microvasive, Boston, MA; Olympus, Tokyo, Japan). The entire esophagus was imaged, and an image was selected at a point where the varices appeared the largest. This image was digitized, and the sum of the CSA of all of the varices was measured (Image Pro Plus, Silver Springs, MD) by an investigator blinded to the patients' clinical status. The follow-up time for each patient was calculated (time to first bleed, time to liver transplantation, time to death, or time to the end of study). The Cox Proportional Hazards Model was used to determine if there was a significant difference between the sums of the CSA in the patients who bled compared with those who did not bleed. An OR was calculated to determine the risk of future variceal bleeding based on the sum of the CSA as measured in cm(2)/month. Positive and negative predictive values were calculated for future variceal bleeding. RESULTS: Six of 28 patients (21%) experienced esophageal variceal bleeding on follow-up. The mean CSA +/- SEM of the sum of the esophageal varices in these patients was 0.77 cm(2) +/- 0.31 cm(2) (range 0.07-2.09 cm(2)). The mean time to bleeding was 15.5 months +/- 4.95 months (range 1-29 months). Twenty-two of 28 patients (79%) did not experience variceal bleeding. The mean CSA +/- SEM of the sum of the varices in these patients was 0.36 cm(2) +/- 0.08 cm(2) (range 0.02-1.19 cm(2)). The mean time to follow-up was 35.7 months +/- 6.69 months (range 1.2-103.2 months). The sum of the CSA between the patients who bleed and those who did not bleed was significantly different at the p < 0.018 level. The OR for the risk of variceal bleeding for each one cm(2) difference in the sum of the CSA per month was 6.34. Using a cutoff of 0.45 cm(2), the sensitivity and specificity for future variceal bleeding was 83% and 75%, respectively. CONCLUSIONS: There is a significant difference (p < 0.018) in the sum of the esophageal variceal CSA between those patients who will experience future variceal bleeding and those who will not. There is a 76-fold increase per year in the risk of future variceal bleeding for each one cm(2) increase in variceal CSA. Using a cutoff value for the CSA of 0.45 cm(2), the sensitivity and specificity for future variceal bleeding above and below this point is 83% and 75%, respectively. PMID- 12591069 TI - Antiphospholipid antibodies before and after liver transplantation. AB - OBJECTIVE: The aim of this study was to determine whether liver transplantation of patients with antiphospholipid antibodies (APA) is 1) adversely affected with vascular thrombosis and 2) whether such antibodies persist post transplantation. METHODS: Twelve patients with APA awaiting transplant were identified and characterized biochemically and immunologically. Each had the level of APA determined using commercially available enzyme-linked immunoassay kits before, during, and after liver transplantation. RESULTS: No patient in this series experienced a transplant-related vascular thrombosis. The titer of APA fell to levels at or below those present in normals and remained low in two of 12 or undetectable in 10 of 12 patients 1 yr after liver transplantation. CONCLUSIONS: We reached the following conclusions: 1) Antiphospholipid positivity does not identify patients at high risk for post-transplant vascular thrombosis. 2) The levels of antiphospholipid present in sera pretransplant fell during transplantation and remained low or undetectable 1 month and 1 yr post transplantation. PMID- 12591070 TI - Hepatic ATP reserve and efficiency of replenishing: comparison between obese and nonobese normal individuals. AB - OBJECTIVE: Although obesity-associated fatty liver disease is emerging as one of the most common diseases in hepatology practice, it is unclear why liver disease prevalence increases with obesity. Because impaired energy homeostasis enhances the susceptibility of hepatocytes to injury, the aim of this study was to determine whether increased body mass index (BMI) is associated with decreased basal hepatic adenosine triphosphate (ATP) stores or impaired recovery from fructose-induced hepatic ATP depletion. METHODS: Hepatic ATP stores were assessed by nuclear magnetic resonance spectroscopy in 19 healthy subjects with varying BMI. After obtaining the baseline spectra, 0.5 ml/kg of 50% fructose solution was administered to all subjects to deplete the ATP reserve, and follow-up nuclear magnetic resonance spectra was obtained at 5-min intervals for the ensuing hour. AST and ALT were determined at 24 h to assess whether ATP depletion caused any appreciable hepatocyte injury. RESULTS: Among the 19 subjects who participated in the study, five had BMI of < or =25, seven had BMI between 25-30, and seven had BMI of >30. The baseline ATP content was inversely related to BMI (correlation coefficient -0.63, p = 0.02), decreasing steadily with increasing BMI. Fructose injection decreased hepatic ATP stores in all subjects and did not increase transaminases in anyone. Neither the postfructose ATP nadir values nor the extent of ATP recovery correlated with BMI. CONCLUSIONS: Reduced hepatic ATP stores are more prevalent in overweight and obese subjects than in lean subjects. However, a cause-effect relationship between these abnormalities was not demonstrated by our study. PMID- 12591071 TI - Outcomes of colorectal cancer in the United States: no change in survival (1986 1997). AB - OBJECTIVES: The past decade has witnessed important advances in colorectal cancer (CRC) screening and treatment. To assess the potential impact of these advances on CRC survival and incidence, we evaluated recent temporal trends in the United States. METHODS: Using the nine population-based cancer registries that constitute the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute, we identified primary CRCs diagnosed between 1986 and 1997. Temporal changes were evaluated for the periods 1986-1988, 1989-1991, 1992 1994, and 1995-1997. Age-adjusted incidence and relative 1-, 3-, and 5-yr survival were calculated, along with 95% CI. RESULTS: We identified 144,284 individuals with CRC during 1986-1997, 51.5% of whom were men. In individuals of white ethnicity, the age-adjusted incidence rates for rectosigmoid and left CRC fell over time. In addition, in white men (but not in white women), the age adjusted incidence rates for right CRC declined. In individuals of African American/black ethnicity, the age-adjusted incidence rates for rectosigmoid and right CRC showed no significant change over time. For all sites, no significant differences in survival were seen over time in whites or blacks. CONCLUSIONS: The decrease in age-adjusted incidence rates over time is consistent with a decrease in CRC burden, especially among whites. However, the lack of improvement in survival over time in whites and blacks indicates that despite advances in screening and treatment, to date, no survival benefit has occurred. PMID- 12591072 TI - Clinical significance of serum levels of E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 in gastric cancer patients. AB - OBJECTIVES: This study evaluated serum concentrations of soluble cell adhesion molecules in patients with gastric cancer and in healthy control subjects. Our objectives were to correlate these levels with clinicopathological features, established tumor markers, and patient survival, and to assess changes in serum levels of cell adhesion molecules after tumor surgery. METHODS: The serum concentrations of the adhesion molecules E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) were investigated by ELISA in 57 gastric cancer patients, both before and 7 days after surgery, and in 47 healthy control subjects. RESULTS: Preoperative serum concentrations of ICAM-1 and VCAM-1 in gastric cancer patients were significantly higher when compared with those of healthy controls, whereas there were no differences regarding serum E-selectin levels. Serum levels of E-selectin, ICAM 1, and VCAM-1 correlated significantly with each other. There was a significant association between preoperative levels of all three adhesion molecules and disease stage, gastric wall invasion, lymph node involvement, and presence of distant metastases. Their concentrations decreased significantly after radical resection of the tumor, whereas they remained almost unchanged in patients with unresectable disease. Elevated preoperative serum levels of E-selectin, ICAM-1, and VCAM-1 levels were found in 24.6%, 33.3%, and 28.1% of patients, respectively. Elevated levels of all three molecules were significant prognostic factors for patient survival but not independent of disease stage. CONCLUSIONS: These findings suggest that serum concentrations of E-selectin, ICAM-1, and VCAM 1 may reflect tumor progression and metastasis, and may be clinically useful. PMID- 12591073 TI - Case-control study of screening colonoscopy in relatives of patients with colorectal cancer. AB - OBJECTIVES: The efficacy of colonoscopic screening and polypectomy for the prevention of colorectal cancer (CRC) is well accepted but has never been documented in a prospective, controlled study. Screening by sigmoidoscopy has been found to reduce mortality from cancer of the rectum and distal colon. Case control studies provide an alternative method for determining the efficacy of screening methods. METHODS: Between 1998 and 2000, a total of 40 subjects were found to have CRC (study group) and 160 had a normal colon (control group) among asymptomatic individuals participating in a screening colonoscopy program for a high-risk population of first-degree relatives of CRC patients. We compared these groups for screening by fecal occult blood testing, flexible sigmoidoscopy, barium enema, and colonoscopy in the 10-yr period before the index colonoscopy. RESULTS: Screening colonoscopy was performed in only 2.5% of the case subjects and 48.7% of controls (p < 0.0001), and all screening procedures in 12.5% and 73.7%, respectively (p < 0.0001). A statistically significant difference was also found for screening with fecal occult blood test, but not for flexible sigmoidoscopy or barium enema. Significant adenomatous polyps >1 cm in diameter were detected and removed in 19% of the control group within 10 yr of the index colonoscopy. Six (15%) of the patients in the study group died of CRC. CONCLUSIONS: Screening by colonoscopy can prevent progression to CRC from adenomatous polyps and may reduce the mortality associated with this devastating disease. PMID- 12591074 TI - Gastric mucosa-associated lymphoid tissue lymphoma with adult T cell leukemia/lymphoma cell infiltration. case report. AB - This is the first case report of gastric mucosal-associated lymphoid tissue lymphoma with adult T cell leukemia/lymphoma (ATLL) cell infiltration. A 43-yr old Japanese woman who was seropositive for antihuman T cell leukemia virus-I antibody complained of epigastric pain in April, 1996. Endoscopy showed gastric ulcers in the antrum. Biopsy specimens showed Helicobacter pylori infection. Her symptoms were relieved by treatment with ranitidine. In March, 1998, she complained of epigastric pain and abdominal fullness. Smears of peripheral blood revealed atypical lymphocytes with nuclear irregularity, consistent with ATLL cells. She was diagnosed to have ATLL. Endoscopy revealed multiple gastric ulcers in the antrum and the angle. Biopsy specimens demonstrated small centrocyte-like cells forming lymphoepithelial lesions, with infiltrations of large atypical lymphoid cells of ATLL. On immunohistochemical staining, the small centrocyte like cells were positive for B cell markers (L26, CD20), whereas the large atypical lymphoid cells were positive for T cell marker (UCHL-1, CD45RO). Her findings were attributed to gastric mucosal-associated lymphoid tissue lymphoma with gastric involvement with ATLL. PMID- 12591075 TI - Current biochemical studies of non-alcoholic fatty liver disease (NAFLD) and non alcoholic steatohepatitis (NASH) suggest a new therapeutic approach. PMID- 12591076 TI - The road to resistance: antibiotics as growth promoters for animals. PMID- 12591077 TI - Problems with NASH? HAIR is some help. PMID- 12591078 TI - Re: Lin and Gray - immunoproliferative small intestinal disease. PMID- 12591079 TI - Nontropical and tropical duodenal mucosa. PMID- 12591080 TI - Diffuse mucosal hemorrhage after air insufflation during endoscopy for PEG placement. PMID- 12591081 TI - Treatment of patients with Crohn's disease and concomitant chronic hepatitis C with a chimeric monoclonal antibody to TNF. PMID- 12591082 TI - The administration of gliadin downregulates the intraepithelial lymphocytosis induced by a rectal gluten challenge. PMID- 12591083 TI - Valsartan-induced hepatotoxicity in a HBs-Ag-Positive patient. PMID- 12591084 TI - Traveler's constipation. PMID- 12591085 TI - Crohn's disease involving the gallbladder. case report and review of the literature. PMID- 12591086 TI - Granulocytapheresis for Crohn's disease: a report on seven refractory patients. PMID- 12591087 TI - Intestinal obstruction after ingestion of a beer-filled condom at the Munich Octoberfest. PMID- 12591089 TI - N-terminal truncation of human alpha1D-adrenoceptors increases expression of binding sites but not protein. AB - The role of the N-terminus of human alpha(1D)-adrenoceptors was examined by deleting the first 79 amino acids (Delta(1-79)) and epitope-tagging to facilitate immunoprecipitation and detection. Following transfection into HEK293 cells, 6- to 13-fold increases in the density of specific [125I]BE 2254 binding sites were observed for both tagged and untagged Delta(1-79)alpha(1D)- compared to full length alpha(1D)-adrenoceptors, while agonist and antagonist affinities remained unchanged. In contrast, immunoprecipitation of tagged receptors showed that full length alpha(1D)-adrenoceptor protein was at least twice as abundant as Delta(1 79)alpha(1D)-adrenoceptor protein. Photoaffinity labelling with [125I]arylazidoprazosin showed much more intense labelling of tagged Delta(1 79)alpha(1D)- than of full-length alpha(1D)-adrenoceptors. Substantial N-linked glycosylation of tagged Delta(1-79)alpha(1D)-adrenoceptors was observed, although full-length alpha(1D)-adrenoceptors contain two consensus glycosylation sites but are not glycosylated. These results suggest that N-terminal truncation of alpha(1D)-adrenoceptors enhances processing of a binding competent form in HEK293 cells; and show a clear dissociation between abundance of receptor protein and density of receptor binding sites. PMID- 12591090 TI - Inhibition by fluoroquinolones of K(+) currents in rat dissociated hippocampal neurons. AB - The effects of four fluoroquinolones (sparfloxacin, fleroxacin, ofloxacin and levofloxacin) on K(+) currents were investigated in pyramidal neurons acutely isolated from rat hippocampus, to evaluate their relative potencies for inhibiting these channels. Using patch-clamp electrophysiological techniques, we found that all four compounds inhibited the delayed rectifier K(+) current (I(K)), but with different potencies. Sparfloxacin was the most potent compound, displaying an IC(50) value of 6.44 x 10(-4) M, followed by fleroxacin, ofloxacin and levofloxacin, their IC(50) values being 7.09 x 10(-3), 8.42 x 10(-3) and 1.10 x 10(-2) M, respectively. In contrast, the fast transient K(+) current (I(A)) was blocked only by sparfloxacin (IC(50)=2.86 x 10(-3) M) and fleroxacin (IC(50)=4.38 x 10(-3) M), but not by ofloxacin and levofloxacin even at concentrations up to 1 mM. The K(+) current inhibition was reversible after washout of the compounds. Further study is needed to clarify the possible involvement of this novel action in the adverse effects of fluoroquinolones in the central nervous system (CNS). PMID- 12591091 TI - Atorvastatin inhibits expression of minichromosome maintenance proteins in vascular smooth muscle cells. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors have been reported to inhibit vascular smooth muscle cell growth, a key event in the pathogenesis of proliferative vascular diseases. The mechanism by which HMG-CoA reductase inhibitors exert their antiproliferative activity is not fully understood, especially their effect on DNA replication. We therefore investigated the effects of atorvastatin on minichromosome maintenance (MCM) protein 6 and 7 expression in vascular smooth muscle cells, two proteins essential for initiation of DNA replication. Stimulation of quiescent rat aortic vascular smooth muscle cells with fetal bovine serum induced MCM6 and MCM7 protein and mRNA expression, which was potently attenuated by atorvastatin in a dose-dependent fashion. Mevalonate completely abrogated the inhibitory effect on serum-induced MCM6 and MCM7 expression, demonstrating that biosynthesis of isoprenoids was likely the specific pathway blocked by atorvastatin. Transient transfection experiments revealed that atorvastatin inhibited MCM6 and MCM7 promoter activity, implicating a transcriptional mechanism. The MCM6 and MCM7 promoters contain several E2F sites critical for their transcriptional activation. Activity of a luciferase reporter plasmid containing four E2F elements was also strongly inhibited by atorvastatin. The inhibitory effect of atorvastatin on MCM6 and MCM7 was reversed by adenoviral-mediated overexpression of E2F, indicating that their downregulation by atorvastatin involves an E2F-dependent mechanism. These findings demonstrate that MCM proteins play an essential role during the proliferation of vascular smooth muscle cells and may provide a novel therapeutic target for proliferative vascular diseases. Inhibition of MCM6 and MCM7 expression by blocking E2F function may contribute importantly to the inhibition of vascular smooth muscle cell DNA synthesis by atorvastatin. PMID- 12591092 TI - Activation of muscarinic m5 receptors inhibits recombinant KCNQ2/KCNQ3 K+ channels expressed in HEK293T cells. AB - A variety of G-protein-coupled receptors regulate membrane excitability via M type K(+) current (M-current) modulation. Muscarinic m1 and m3 acetylcholine receptors have both been implicated in the modulation of M-current. The muscarinic m5 receptor, like muscarinic m1 and m3 receptors, couples to phospholipase C via a pertussis toxin-insensitive G protein. Since a number of other receptors which activate phospholipase C also modulate M-current, we investigated if muscarinic m5 receptors could modulate recombinant M-type (KCNQ2/KCNQ3) K(+) channels after heterologous expression in human embryonic kidney (HEK) 293T cells. Application of Oxo-tremorine M to HEK293T cells expressing muscarinic m1, m3, or m5 receptors produced a similar robust inhibition of M-current, whereas muscarinic m2 and m4 receptor stimulation was without effect. Muscarinic m1, m3, or m5 receptor stimulation decreased the deactivation time constants of M-current at -50 mV. The inhibition of M-current by stimulation of muscarinic m1, m3, or m5 receptors was insensitive to overnight treatment with pertussis toxin or cholera toxin, which interfere with G(i/o) and G(s) G-protein signaling. These data suggest that muscarinic m1, m3, and m5 receptors inhibit M-channels via the activation of a common G protein. PMID- 12591093 TI - alpha2C-Adrenoceptor blockade by clozapine and other antipsychotic drugs. AB - The noradrenergic system may play a role in antipsychotic modulation of schizophrenia symptoms. Therefore, the antagonistic potencies of the antipsychotics clozapine, chlorpromazine, risperidone, olanzapine, haloperidol, quetiapine, ziprasidone, iloperidone and aripiprazole were quantified using cell lines expressing the recombinant human alpha(2C)-adrenoceptor, alpha(2A) adrenoceptor, or dopamine D(2L) receptor. The alpha(2)-adrenoceptor antagonists, yohimbine and idazoxan, were also tested. Alterations in cAMP were measured as changes in luminescence. In the alpha(2A)-adrenoceptor cell line, the agonist 5 bromo-6-(2-imidazolin-2-ylamino)quinoxaline (UK14,304) induced a concentration dependent increase in luminescence. In cell lines expressing alpha(2C) and D(2L) receptors, agonists induced a concentration-dependent reduction in luminescence. Yohimbine and idazoxan were the most potent alpha(2A)-adrenoceptor antagonists, yohimbine and iloperidone were the most potent alpha(2C)-adrenoceptor antagonists, and haloperidol and olanzapine were the most potent dopamine D(2) receptor antagonists. Clozapine had the highest alpha(2C)/D(2) selectivity, and iloperidone the highest alpha(2C)/alpha(2A) ratio. It is hypothesised that alpha(2C)-adrenoceptor blockade contributes to improvement of cognitive function. PMID- 12591094 TI - Characterization of aliphatic, cyclic, and aromatic N-terminally "capped" His-D Phe-Arg-Trp-NH2 tetrapeptides at the melanocortin receptors. AB - The melanocortin system is implicated in multiple physiological pathways including pigmentation, inflammation, erectile function, feeding behavior, energy homeostasis, weight homeostasis, and exocrine gland function, just to list a few. The endogenous agonists for the melanocortin receptors include the gene transcripts derived from the proopiomelanocortin gene and include the core tetrapeptide His-Phe-Arg-Trp sequence postulated to be important for melanocortin receptor selectivity and stimulation. Posttranslational processing of the proopiomelanocortin derived agonists results in the N-terminal acetylation and C terminal amidation of alpha-melanocyte stimulation hormone (alpha-MSH). In this study we generated 25 N-terminally "capped" tetrapeptides containing the core sequence X-His-D-Phe-Arg-Trp-NH(2) and pharmacologically characterized them at the mouse melanocortin MC(1) receptor, melanocortin MC(3) receptor, melanocortin MC(4) receptor, and melanocortin MC(5) receptor. The N-terminal "capping" groups consisted of linear, cyclic, or aromatic moieties and all resulted in full agonist activity at the melanocortin receptors examined in this study. Increasing aliphatic chain length increased potency of the tetrapeptide derivatives, with the addition of octanoyl capping group resulting in 70- to 110-fold increased tetrapeptide potency at the melanocortin MC(1) receptor (EC(50)=0.4 nM), melanocortin MC(3) receptor (EC(50)=4.0 nM), and melanocortin MC(4) receptor (EC(50)=0.4 nM) while only enhancing potency at the melanocortin MC(5) receptor (EC(50)=0.8 nM) by 8-fold, compared to the tetrapeptide His-D-Phe-Arg-Trp-NH(2). This octanoyl derivative surprisingly resulted in a 14-fold greater potency than alpha-MSH (EC(50)=5.4 nM) at the mouse melanocortin MC(4) receptor implicated in feeding behavior and obesity. The 3,3,3-triphenylpropionyl derivative resulted in greater than 14 microM agonist potencies at the melanocortin MC(1) receptor, melanocortin MC(3) receptor, and melanocortin MC(4) receptor and possessed a 140 nM agonist EC(50) value at the melanocortin MC(5) receptor. This 3,3,3 triphenylpropionyl-His-D-Phe-Arg-Trp-NH(2) peptide is a 100-fold selective agonist for the melanocortin MC(5) receptor, versus the other melanocortin receptors studied herein, and is the first melanocortin MC(5) receptor selective tetrapeptide derivative reported to date with nanomolar potency. PMID- 12591095 TI - A synthetic ceramide analog ameliorates spatial cognition deficit and stimulates biosynthesis of brain gangliosides in rats with cerebral ischemia. AB - A synthetic ceramide analog, L-threo-1-phenyl-2-decanoylamino-3-morpholino-1 propanol (L-PDMP) upregulates ganglioside biosynthesis in several cell lines. In cultured cortical neurons, neurotrophic effects of L-PDMP on neurite outgrowth and synaptic activity were demonstrated. In addition, it was found that L-PDMP could ameliorate the spatial cognition deficit in rats with ischemia. To elucidate this effect, we evaluated the effect of L-PDMP on brain ganglioside biosynthesis and its therapeutic efficacy against spatial cognition deficit in rats made ischemic. Rats were trained for 2 weeks, using an 8-arm radial maze task, and then forebrain ischemia was induced. L-PDMP was injected i.p. at 40 mg/kg twice a day starting from day 1 or 3 after ischemia induction for 6 or 4 days, respectively. The first study showed significantly reduced spatial cognition deficit at 12 h after the final drug administration, and L-PDMP tended to attenuate apoptosis in hippocampal CA1. To examine the effect of L-PDMP on brain ganglioside biosynthesis, N-[3H]acetyl-D-mannosamine was infused into the lateral ventricle via an injection cannula at 12 h after the final drug administration. After 4 h, the brain gangliosides were purified and analyzed. Upregulation of ganglioside biosynthesis by L-PDMP was observed on days 3 and 5 after ischemia. These results are an indication that L-PDMP may ameliorate spatial cognition deficit by upregulating ganglioside biosynthesis in ischemic brain. PMID- 12591096 TI - Acute effect of simultaneous administration of tryptophan and ethanol on serotonin metabolites in the locus coeruleus in rats. AB - Using the microdialysis method, we investigated whether the levels of serotonin (5-hydroxytryptamine, 5-HT) and its metabolites, 5-hydroxyindoleacetic acid (5 HIAA) and 5-hydroxytryptophol (5-HTPL), in the locus coeruleus are influenced by tryptophan alone or simultaneous administration of tryptophan and ethanol. Tryptophan (50 mg/kg, i.p.) led to a significant increase in the levels of 5 HIAA, but not 5-HT in the locus coeruleus. However, ethanol (1.25 g/kg) had no effect on the levels of 5-HT and its metabolites. Combined administration of tryptophan and ethanol caused very marked increases in 5-HIAA and 5-HTPL levels in the locus coeruleus. A time lag in the increased 5-HIAA levels between tryptophan alone and tryptophan plus ethanol was observed. Moreover, 5-HIAA levels in the locus coeruleus induced by tryptophan were abolished by microinjection of 5,7-dihydroxytryptamine (150 microg/4 microl) into the dorsal raphe nucleus. Judging from the present results, the serotonergic afferents to the locus coeruleus may originate for about 20-30% from cell bodies located in the dorsal raphe nucleus. Teeth-chattering was significantly detected in the tryptophan plus ethanol-treated rats when compared with the tryptophan-treated rats, but not in the saline-treated controls. These results may suggest that the increased levels of 5-HIAA and 5-HTPL in the locus coeruleus induced by tryptophan are potentiated by ethanol, and that these levels are partly responsible for behavioral activation. PMID- 12591097 TI - Increase of glial fibrillary acidic protein and S-100B in hippocampus and cortex of diabetic rats: effects of vitamin E. AB - Glial interactions with neurones play vital roles during the ontogeny of the nervous system and in the adult brain. Physical and metabolic insults cause rapid changes in the glial cells and this phenomenon is called reactive gliosis. One of the important events during astrocyte differentiation is the increased expression of glial markers, glial fibrillary acidic protein (GFAP) and S-100B protein. Diabetes mellitus is the most common serious metabolic disorder, which is characterised by functional and structural changes in the peripheral as well as in the central nervous system. In the present study, we aimed to investigate glial reactivity in hippocampus, cortex and cerebellum of streptozotocin-induced diabetic rats by determining the expression of GFAP and S-100B and also to examine the protective effects of vitamin E against gliosis. Western blotting showed increases in total and degraded GFAP content and S-100B protein expression in brain tissues of diabetic rats compared with those of controls. In addition, there was a significant increase in lipid peroxidation in these brain regions of diabetic rats. Both glial markers and lipid peroxidation levels were reversed by vitamin E administration. These findings indicate that streptozotocin-induced diabetes alters degradation and production of GFAP and S-100B, which are markers of reactive astrocytosis. Thus, determination of GFAP and S-100B may provide a relevant marker in the central nervous system for studying neurodegenerative changes in experimental diabetes mellitus. This study also suggests that the gliosis that occurs in diabetes mellitus is mediated, at least indirectly, by free radical formation and antioxidants may prevent reactive gliosis possibly by reducing damaging effects of reactive oxygen species in the central nervous system. PMID- 12591098 TI - Group I metabotropic glutamate receptors modulate glutamate and gamma aminobutyric acid release in the periaqueductal grey of rats. AB - In this study, we investigated the effects of group I metabotropic glutamate (mglu) receptor ligands on glutamate and gamma-aminobutyric acid (GABA) extracellular concentrations at the periaqueductal grey level by using in vivo microdialysis. An agonist of group I mglu receptors, (S)-3,5 dihydroxyphenylglycine [(S)-3,5-DHPG, 1 and 2 mM], as well as a selective agonist of mglu(5) receptors, (RS)-2-chloro-5-hydroxyphenylglycine (CHPG, 2 and 4 mM), both increased dialysate glutamate and GABA concentrations. 7 (Hydroxyimino)cyclopropa-[b]-chromen-1alpha-carboxylate ethyl ester (CPCCOEt, 1 mM), a selective mglu(1) receptor antagonist, and 2-methyl-6 (phenylethynyl)pyridine (MPEP, 0.5 mM), a selective mglu(5) receptor antagonist, perfused in combination with DHPG, antagonized the effect induced by DHPG on the extracellular glutamate and GABA concentrations. MPEP (0.5 mM), perfused in combination with CHPG, antagonized the increased glutamate and GABA extracellular levels induced by CHPG. MPEP (1 mM) decreased the extracellular concentrations of glutamate but did not modify the dialysate GABA concentrations. Moreover, as the intra-periaqueductal grey perfusion of (RS)-3-(2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid [(RS)-CPP, 100 microM], a selective N-methyl-D-aspartate (NMDA) glutamate receptor antagonist, did not change the extracellular concentrations of glutamate, this suggests that the MPEP-induced decrease in glutamate is not a consequence of NMDA receptor blockade. These data show that group I mglu receptors in the periaqueductal grey may modulate the release of glutamate and GABA in awake, freely moving rats. In particular, mglu(5), but not mglu(1), receptors seem to be functionally active on glutamate terminals. PMID- 12591099 TI - Atipamezole, an alpha2-adrenoceptor antagonist, augments the effects of L-DOPA on evoked dopamine release in rat striatum. AB - The effects of atipamezole, an alpha(2)-adrenoceptor antagonist, L-3,4 dihydroxyphenylalanine (L-DOPA) and the combination of these drugs on dopamine overflow were studied in dopaminergic presynaptic terminals of rat caudate and nucleus accumbens. Dopamine overflow evoked by 100 pulses of electrical stimulation of the medial forebrain bundle at a low (20 Hz) and high (50 Hz) frequency was measured by in vivo voltammetry. L-DOPA (15 mg/kg) increased dopamine overflow in the caudate nucleus, but this dose had no effects in the nucleus accumbens. Atipamezole (300 microg/kg) had no effects on its own on dopamine overflow, but it did increase the size of the readily releasable storage pool and the effects of L-DOPA treatment in both structures. The combination of the drugs increased dopamine overflow to a larger extent at high compared to low stimulation frequencies. We conclude that the rat caudate nucleus is more sensitive than the nucleus accumbens to the effects of L-DOPA, and the effects of L-DOPA treatment might be effectively enhanced by antagonism of alpha(2) adrenoceptors. PMID- 12591100 TI - The presynaptic modulation of corticostriatal afferents by mu-opioids is mediated by K+ conductances. AB - Population spikes associated with the paired pulse ratio protocol were used to measure the presynaptic inhibition of corticostriatal transmission caused by mu opioid receptor activation. A 1 microM of [D-Ala(2), N-MePhe(4), Gly-ol(5)] enkephalin (DAMGO), a selective mu-opioid receptor agonist, enhanced paired pulse facilitation by 44+/-8%. This effect was completely blocked by 2 nM of the selective mu-receptor antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-NH (CTOP). Antagonists of N- and P/Q-type Ca(2+) channels inhibited, whereas antagonists of potassium channels enhanced, synaptic transmission. A 1 microM of omega-conotoxin GVIA, a blocker of N-type Ca(2+) channels, had no effect on the action of DAMGO, but 400 nM omega-agatoxin TK, a blocker of P/Q-type Ca(2+)-channels, partially blocked the action of this opioid. However, 5 mM Cs(2+) and 400 microM Ba(2+), unselective antagonists of potassium conductances, completely prevented the action of DAMGO on corticostriatal transmission. These data suggest that presynaptic inhibition of corticostriatal afferents by mu-opioids is mediated by the modulation of K(+) conductances in corticostriatal afferents. PMID- 12591101 TI - New benzimidazole derivatives: selective and orally active 5-HT3 receptor antagonists. AB - The synthesis of new 5-HT(3) receptor antagonists is an interesting field of research because of their wide therapeutic use. The aim of this work is to functionally characterise a new series of benzimidazole derivatives previously described. These compounds bind to 5-HT(3) receptors and have been evaluated using in vitro (rat tunica muscularis mucosae) and in vivo tests (Bezold-Jarisch reflex in rat and gastrointestinal motility and spontaneous motility in mice). Ondansetron and 1-[4-amino-5-chloro-2-(3,5-dimethoxyphenil)methyloxy]-3-[1-[2 methylsulfonylamino]piperidin-4-yl]propan-1-one hydrochloride (RS 39604) were used as well known 5-HT(3) and 5-HT(4) receptor antagonists. These benzimidazole derivatives have proved to be 5-HT(3) receptor antagonists. Interestingly, they are as active as ondansetron when they are intraperitoneally (i.p.) or orally (p.o.) administered and, in mice, they seem to induce fewer behavioural changes at similar effective doses than does ondansetron. The present results confirm the usefulness of the previously proposed pharmacophore and justify the interest in these new benzimidazole derivatives. PMID- 12591102 TI - Effects of morphine on formalin-induced nociception in rats. AB - This study focused on the antinociceptive action of morphine in the formalin test in rats. Formalin-induced behaviour is characterised by two phases relevant to acute and tonic pain. Morphine (1-6 mg/kg) was administered systemically before or after the early phase, and its ability to affect the late phase was investigated. Inhibitory effects of morphine (3 mg/kg) injected immediately after the early phase were significantly stronger (32+/-9%) compared to the preemptive administration (84+/-29%, relative to saline-treated controls, 5% formaldehyde). It appears that some neural and/or behavioural changes during the early phase limit effects of morphine on the late phase. Furthermore, manipulation of stimulation intensity (2% vs. 5% formaldehyde) significantly affected the ability of morphine (3 mg/kg) to suppress early (55+/-7% and 76+/-10%, respectively) but not late phase of formalin-induced behaviours. These results agree with the previous demonstrations on the effects of acute nociceptive stimulation intensity on analgesic potency of opiate drugs. Thus, the present study revealed two factors that affect the potency of morphine in formalin test: administration regimen and formalin concentration. PMID- 12591103 TI - Ethanol improves short-term social memory in rats. Involvement of opioid and muscarinic receptors. AB - Some human and animal studies have demonstrated enhancement of memory processes when ethanol was administered immediately after training and subjects were later tested in the drug-free state. The aim of this study was to evaluate the effect of acute ethanol administration (0.5, 1.0 and 2.0 g/kg) by intraperitoneal (i.p.) and oral route on short-term memory, using the social recognition test in rats. The actions of scopolamine (0.06 and 0.5 mg/kg, i.p.) and naloxone (1.0 mg/kg, i.p.) and their interaction with ethanol in relation to short-term memory were also studied. The doses of ethanol used did not show any sedative effect, which was assessed by measuring locomotor activity. The results indicate that acute low doses of ethanol (0.5 and 1.0 g/kg, i.p.) improve the short-term olfactory memory in rats in a specific and time-dependent manner, and that this action is, at least in part, related to opioid, but not to muscarinic receptors. In addition, these findings confirm that the social recognition test in rats is a useful and reliable model to investigate short-term memory affected by ethanol. PMID- 12591104 TI - Antiobesity effects of chronic cannabinoid CB1 receptor antagonist treatment in diet-induced obese mice. AB - We determined the effect of a cannabinoid CB1 receptor antagonist (AM-251; N (Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamide) on food intake, body weight and adipose tissue mass in Western diet induced obese (DIO) mice using a chronic, interrupted, oral dosing paradigm. The dosing paradigm was 2 weeks on treatment (treatment 1), 2 weeks off-treatment, followed by 2 weeks on treatment (treatment 2). During treatment 1 and treatment 2, food intake and body weight were reduced after a single dose. At 30 mg/kg/day, anorectic efficacy was maintained through 12 days (treatment 1) and 7 days (treatment 2). Body weight of AM-251-treated mice remained less than vehicle treated mice throughout treatment 1 and treatment 2. Administration of AM-251 reduced inguinal subcutaneous, retroperitoneal and mesenteric adipose tissue mass. Antiobesity effects of AM-251 were lost during the off-treatment period, and hyperphagia was observed in treated animals. With re-initiation of AM-251 treatment, mice again responded to the effects of the compound. These results support the hypothesis that chronic treatment of obese individuals with cannabinoid CB1 receptor antagonists is a viable pharmacologic approach to sustained weight loss. PMID- 12591105 TI - Comparative behavioural effects of typical and atypical antipsychotic drugs in rhesus monkey. AB - Behavioural effects of typical and atypical antipsychotic agents were compared in unrestrained rhesus monkey (Macaca mulatta) living in social colonies. The behaviours were categorized as social, solitary and abnormal. They were studied with the help of video cameras fixed in the observation chamber. The behavioural effects were recorded on videotape and analyzed for significant changes. Chlorpromazine (2.5-10 mg/kg, i.m.), haloperidol (0.01-0.04 mg/kg, i.m.), risperidone (0.05-0.2 mg/kg, p.o.) and clozapine (5-20 mg/kg, p.o.) induced significant alterations in parameters of social and solitary behaviour. Chlorpromazine produced a marked decrease in locomotor activity whereas haloperidol showed marked extrapyramidal effects. Risperidone produced minimal extrapyramidal effects and sedation compared to haloperidol and chlorpromazine. Clozapine had intermediate extrapyramidal effects similar to those of chlorpromazine but it produced hypersalivation and dose-related sedation. Thus, risperidone had advantages over the other antipsychotics used in this study because it did not produce salivation, had minimal extrapyramidal effects and caused less sedation. These antipsychotic drugs produced many behavioural effects in the rhesus monkey that were similar to their clinically observed effects. A study of behavioural effects in the monkey can thus be a useful predictive tool in the preclinical development of new antipsychotics. PMID- 12591106 TI - Effect of a single dose aspirin on platelets in humans with multiple risk factors for coronary artery disease. AB - We sought to assess how one tablet of non-enteric coated aspirin (325 mg) affects human platelets in subjects with risk factors for coronary artery disease. Data from 63 individuals with multiple cardiac risk factors were analyzed. Platelets were assessed twice at baseline (pre-aspirin), and after 3-4 h (post-aspirin). We employed 5 microM epinephrine-induced conventional aggregometry, closure time with epinephrine/collagen cartridge by PFA-100(R) (Dade-Behring), and aspirin response units (ARU) stimulated by propyl gallat with Ultegra (Accumetrics, San Diego, CA, USA) for measuring platelet function. In addition, the expression of platelet receptors was determined by using the following monoclonal antibodies: anti-CD31, CD41, CD42b, CD51/CD61, CD62p, CD63, CD107a, and CD151. Platelet leukocyte formation was detected utilizing dual antibodies for a pan-platelet marker CD151, and CD14, a monocyte/macrophage marker. PAC-1 was used to measure fibrinogen-platelet binding. One pill of aspirin significantly decreased platelet rich plasma (PRP) aggregation (74.18+/-16.75% vs. 24.92+/-8.64%; p<0.0001) and resulted in reduction of the aspirin response units (ARU) (662.24+/-65.65 vs. 451.05+/-69.31; p<0.0001). There was also prolongation of the closure time (194.4+/-25.3 vs. 258.63+/-55.61 s; p<0.0001). High correlation (r(2)=0.73-0.86) between platelet analyzer readings and aggregation was observed. One tablet of aspirin moderately inhibited expression of most surface platelet receptors measured, and such inhibition reached significance (p<0.05) for PAC-1, CD31, CD41, CD42, CD62p, and CD151. We conclude that a single dose of aspirin affects major platelet receptors, presumably directly or indirectly through the inhibition of prostanoids via platelet cyclooxygenase-1 blockade. The Ultegra Analyzer with a novel cartridge seems to be reliable in reflecting aspirins' effects on platelets and could be used in the future in clinical practice for monitoring aspirin therapy. PMID- 12591107 TI - Angiotensin II activates NADPH oxidase in isolated rat hearts subjected to ischaemia-reperfusion. AB - The role of angiotensin II in myocardial ischaemia-reperfusion is not clearly defined. In this respect, the involvement of NADPH oxidase remains to be determined. The aim of this study was 1) to evaluate the cardiac effects of angiotensin AT(1) receptor stimulation in non-ischaemic conditions of perfusion or during ischaemia-reperfusion, and 2) to measure the concomitant activation of NADPH oxidase in isolated rat hearts perfused with angiotensin II and/or Losartan. In non-ischaemic hearts, angiotensin II induced rapid and prolonged vasoconstrictive and negative inotropic effects. Ischaemia-reperfusion increased the mRNA expression of AT(1) and AT(2) receptors. During reperfusion, angiotensin II reduced the incidence of arrhythmias and the lactate dehydrogenase released, and increased NADPH oxidase mRNA expression and enzyme activity. Losartan co administration totally antagonised the effects of angiotensin II. Our study demonstrates that ischaemia-reperfusion induces adaptative cardiac modifications, which allow exogenously added angiotensin II to stimulate myocardial NADPH oxidase through angiotensin AT(1) receptor activation. PMID- 12591108 TI - The effect of chronic renal failure on phosphodiesterase inhibitor-induced relaxation responses in rabbit cavernosal strips. AB - Erectile dysfunction is common in men with chronic renal failure. Previously nitrergic and endothelium-dependent relaxation responses have been shown to be reduced in chronic renal failure rabbits. We have therefore investigated the efficacy of phosphodiesterase inhibitors on the corpora cavernosa obtained from uremic rabbits. Uremia was induced with 5/6 nephrectomy and 4 weeks later cavernosal tissue strips were isolated. The relaxant effect of phosphodiesterase 5 inhibitors, zaprinast (1-300 microM) and sildenafil (0.01-300 microM), phosphodiesterase 3 inhibitor amrinone (1-100 microM) and non-specific phosphodiesterase inhibitor papaverine (1-300 microM) were investigated on phenylephrine (10 microM)-induced tone. We found a shift in the dose-response curve of only phosphodiesterase 5 inhibitors. These results suggest that the decreased production or availability of endogenous nitric oxide in chronic renal failure animals leads to decreased efficacy of phosphodiesterase 5 inhibitors to induce relaxation. PMID- 12591109 TI - Inhibition by ebselen on nitric oxide mediated relaxations in the rat anococcygeus muscle. AB - The effect of 2-phenyl-1,2-benzisoselenazol-3(2H)-one (ebselen) on nitric oxide (NO) mediated responses and NO generation from NO donors were studied in vitro. In precontracted rat isolated anococcygeus muscles, relaxations induced by NO donors, electrical field stimulation and 5-[1-(phenylmethyl)-1H-indazole-3-yl]-2 furanmethanol (YC-1) were significantly inhibited by ebselen (100 microM), whereas responses elicited by papaverine and theophylline were not affected; those by 8-bromo-cyclic-guanosine-monophosphate (8-Br-cGMP) were slightly enhanced. NO generation from NO gas aqueous solution or acidified nitrite was not affected, but that from S-nitroso-N-acetyl-penicillamine (SNAP) was attenuated by ebselen, and the attenuation was reserved by glutathione. Both glutathione and cupric sulphate altered the ultraviolet spectrum of ebselen. These findings suggest that ebselen at high concentrations nonselectively inhibited NO-mediated responses, possibly through inhibiting soluble guanylate cyclase. Ebselen does not appear to directly interact with NO, but it may inhibit NO release from nitrosothiols by a thiol- and/or copper-dependent mechanism. PMID- 12591110 TI - Contractile actions of imidazoline alpha-adrenoceptor agonists and effects of noncompetitive alpha1-adrenoceptor antagonists in human vas deferens. AB - The contractile actions of imidazoline alpha-adrenoceptor agonists were investigated in human vas deferens longitudinal and circular muscle. The effects of phenoxybenzamine were studied in comparison to dibenamine and SZL-49 (4-amino 6,7-dimethoxy-2-quinazolinyl-4-(2-bicyclo[2,2,2]octa-2,5-dienylcarbonyl-2 piperazine), an alkylating prazosin analogue that discriminates between alpha(1H) and alpha(1L)-adrenoceptor subtypes. The imidazoline alpha-adrenoceptor agonist, A-61603 (N-[5-(4,5-dihydro-1H-imidazol-2yl)-2-hydroxy-5,6,7,8 tetrahydronaphthalen-1-yl]methanesulfonamide hydrobromide), was a potent agonist (pD(2); longitudinal muscle 6.9, circular muscle 6.4) and cirazoline a partial agonist (pD(2); longitudinal muscle 6.1, circular muscle 5.1). Oxymetazoline was less effective, indanidine and clonidine were ineffective. SZL-49 produced a differential inhibition of contractions evoked by A-61603 in circular (alpha(1H)) compared to longitudinal (alpha(1L)) muscle and phenoxybenzamine had the opposite effect. Dibenamine inhibited the contractions comparably in both muscle types and analyses of its partial alkylation of receptors yielded identical estimates of equilibrium dissociation constant (pK(d)) for A-61603 in longitudinal (5.82) and circular (5.84) muscle. Receptor occupancy-response relationships revealed that whilst the muscle types are not different in receptor reserves for A-61603, contraction to the potent imidazoline is more efficiently coupled in longitudinal than in circular muscle. This underlies the markedly different responsiveness of the muscle types to cirazoline or oxymetazoline (alpha-adrenoceptor agonists with lower efficacies relative to A-61603). The differential inhibitory actions of phenoxybenzamine and SZL-49 are discussed. PMID- 12591111 TI - The potent melanocortin receptor agonist melanotan-II promotes peripheral nerve regeneration and has neuroprotective properties in the rat. AB - The neurotrophic and neuroprotective potential of the alpha-melanocyte stimulating hormone (alpha-MSH) analog cyclo-[Ac-Nle(4),Asp(5),D Phe(7),Lys(10)]alpha-MSH-(4-10) amide (melanotan-II), a potent melanocortin receptor agonist, was investigated. The sciatic nerve crush model was used as a paradigm to investigate the neurotrophic properties of melanotan-II. Melanotan-II significantly enhanced the recovery of sensory function following a crush lesion of the sciatic nerve in the rat at a dose of 20 microg kg(-1) per 48 h, s.c., but not at a dose of 2 or 50 microg kg(-1). In addition, we observed that melanotan II also possesses neuroprotective properties, as it partially protected the nerve from a toxic neuropathy induced by cisplatin. Thus, the present data for the first time demonstrate the effectiveness of the potent alpha-MSH analog melanotan II in nerve regeneration and neuroprotection. PMID- 12591112 TI - Combined action of vasoactive amines and bradykinin mediates allergen-evoked thermal hyperalgesia in rats. AB - The ability of allergens to induce hyperalgesia in immunoglobulin E (IgE) sensitized rats was investigated. The left hind paws of Wistar rats were sensitized with intraplantar injections of IgE anti-dinitrophenylated bovine serum albumin monoclonal antibody, and challenged with dinitrophenylated bovine serum albumin 24 h later. Allergen challenge yielded rapid thermal hyperalgesia and oedema formation in the ipsilateral paws, both reaching a plateau from 15 min to 3 h, and both diminishing thereafter. Allergen-evoked hyperalgesia was inhibited by intraperitoneal treatment with meclizine or methysergide, histamine and 5-hydroxytryptamine receptor antagonists. There was also sensitivity to local treatment with either bradykinin B(1) or B(2) receptor antagonists, des-Arg(9) [Leu(8)]-bradykinin or D-arginyl-[Hyp3, Thi5, D-Tic7, Oic8]-bradykinin (Hoe 140). Anaphylactic hyperalgesia was mimicked by the combined administration of histamine, 5-hydroxytryptamine and bradykinin at doses which were ineffective when injected alone. This synergistic effect was abolished by treatment with either meclizine, methysergide, Hoe 140 or des-Arg(9)-[Leu(8)]-bradykinin. Our findings show that local thermal hyperalgesia is a feature of allergen-evoked inflammation, and that a synergistic interaction among bradykinin, 5 hydroxytryptamine and histamine plays a critical role in this phenomenon. PMID- 12591113 TI - Pharmacological modulation of interleukin-17-induced GCP-2-, GRO-alpha- and interleukin-8 release in human bronchial epithelial cells. AB - The cytokine interleukin-17 may play a role in the recruitment of airway neutrophils, and interleukin-17 protein is increased in the airways of patients with asthma. In this study, we characterised the effect of interleukin-17 on the release of the neutrophil-recruiting cytokines granulocyte chemotactic protein (GCP)-2, growth-related oncogene (GRO)-alpha and interleukin-8 in human bronchial epithelial (HBE) cells. We also characterised the involvement of mitogen activated protein (MAP) kinases as well as the effect of beta-adrenoceptor and glucocorticoid receptor stimulation and calcineurin and P-glycoprotein inhibition on these epithelial responses to interleukin-17. We found that interleukin-17 (1 1000 ng/ml) increased the release of GCP-2, GRO-alpha and interleukin-8 in a concentration-dependent manner. This interleukin-17-induced release of C-X-C chemokines was sensitive to inhibition of the p38 MAP kinase pathway and to stimulation of glucocorticoid receptors. In contrast, stimulation of beta adrenoceptors increased the release of interleukin-8 and did not markedly alter the release of GCP-2 and GRO-alpha. Inhibition of calcineurin and of P glycoproteins did not exert any substantial effect on the release of C-X-C chemokines. In conclusion, interleukin-17 bears the potential to increase neutrophil recruitment into the airways by releasing several, different C-X-C chemokines, including GCP-2, GRO-alpha and interleukin-8 in human bronchial epithelial cells. Inhibition of the p38 MAP kinase pathway and glucocorticoid receptor stimulation constitute two credible therapeutic strategies against this interleukin-17-induced release of neutrophil-recruiting cytokines. PMID- 12591114 TI - Increased masking response to light after ablation of the visual cortex in mice. AB - Mice are known to suppress their wheel running when given a pulse of light in the night (masking response). The amount of suppression can be quantified; the response varies with the level of irradiance used during the light pulse. After ablation of the visual cortex, mice suppressed their activity more than sham operated controls. In addition, the lesioned animals responded to lower levels of irradiance than controls. It is suggested that the visual cortex is not needed for the suppression of locomotor activity after a light pulse. Nevertheless it exerts an inhibitory influence on the masking response to light mediated by an irradiance detection system. When this inhibition is removed, even though pattern vision is lost, masking responses to ambient level of light are enhanced. PMID- 12591115 TI - Calcium 'leak' through somatic L-type channels has multiple deleterious effects on regulated transmitter release from an invertebrate hair cell. AB - Using an identified synapse in the nervous system of the mollusc Hermissenda, the influence of somatic calcium accumulation on regulated synaptic transmission was investigated. Hair cells in Hermissenda project onto postsynaptic B photoreceptors where they mediate inhibitory postsynaptic potentials (IPSPs). Intracellular recordings in combination with bath perfusion of calcium channel modulators indicated that L-type channels were present on the hair cell soma but not on the terminal branches. In contrast, P/Q and an unidentified channel type (similar to N-type channels) contributed additively to transmitter release from the hair cell. Antibodies raised against rat brain channel proteins detected L- (alpha1(C)) and P/Q-type (alpha1(A)) channels in lysates of the Hermissenda nervous system, indicating a homology between the Hermissenda channels and their mammalian counterparts. To mimic somatic calcium channel 'leak', hair cells were exposed to the L-type channel agonist +/-BAY K 8644. Exposure to +/-BAY K 8644 resulted in a rapid (<2 min) increase (40%) in the amplitude of the spike after hyperpolarization in the hair cell, and was associated with a reduction in evoked firing frequency. This reduction in rate of discharge induced a proportional decrease in the amplitude of compound IPSPs recorded in the postsynaptic B photoreceptors. From Fura-2 emissions we determined that +/-BAY K 8644 induced a rapid (<2 min) and persistent increase (70%) in somatic calcium concentration, followed by a slower elevation of calcium in the medial axon (>30 min) and subsequently in the terminal branches (>40 min), suggesting that excessive somatic calcium had diffused or induced a propagation along the axon. Corresponding with a 56% rise in terminal calcium (50-60 min post agonist), postsynaptic potentials declined to 70% of baseline amplitude. These results suggest that prolonged somatic L-channel 'leak' can interfere with regulated transmitter release, both by reducing the rate of presynaptic discharge and by promoting terminal calcium accumulation that may oppose transmitter release. Such effect may have implications for the age-related learning deficits that often accompany somatic calcium 'leak'. PMID- 12591116 TI - Opioid modulation of taste responses in the nucleus of the solitary tract. AB - Gustatory processing within the medulla is modulated by a number of physiologic and experiential factors. Several neurotransmitters, including excitatory amino acids, GABA, and substance P, are involved in synaptic processing within the rostral portion of the nucleus of the solitary tract (NST). Endogenous opiates have been implicated in the regulation of feeding behavior and in taste palatability and gustatory responses in the parabrachial nuclei are reduced by systemic morphine. In the present experiments, extracellular recording of neuronal activity within the NST in response to taste input was combined with local microinjection of met-enkephalin (Met-ENK) and naltrexone (NLTX) to determine the effect of these agents on gustatory activity. The anterior tongue was stimulated with anodal current pulses to determine the time course of drug action (n=85 cells) and with prototypical taste stimuli (0.032 M sucrose, NaCl, and quinine hydrochloride, and 0.0032 M citric acid) to investigate the effects of these opioid compounds on taste-evoked responses (n=80 cells). Among these 165 taste-responsive neurons in the NST, the activity of 39 (23.6%) was suppressed by Met-ENK. These effects were dose-dependent and blockable by NLTX, which alone was without effect, suggesting that opiates do not maintain a tonic inhibitory influence. Immunohistochemical experiments demonstrated both micro - and delta opioid receptors within the gustatory portion of the NST; previous studies had shown numerous fiber terminals containing Met-ENK. These data suggest that endogenous opiates play an inhibitory role in gustatory processing within the medulla. PMID- 12591117 TI - Osmotic regulation of angiotensin AT1 receptor subtypes in mouse brain. AB - Experiments were performed to study angiotensin (Ang) AT1a and AT1b mRNA expression in mice, including, examination of brain distribution and the effect of salt loading. In situ hybridization (ISH) methods showed that the pattern of mRNA expression was identical for AT1a and AT1b, with cellular labeling in rostral forebrain, hypothalamus and brainstem. Receptor mRNAs were concentrated in brain regions involved in the regulation of electrolyte and cardiovascular balance. Immunocytochemistry with AT1 specific antisera showed a pattern that was consistent with the ISH. Reverse transcriptase-polymerase chain reaction (RT-PCR) of hypothalamus and pituitary verified the presence of both AT1a and AT1b mRNA. Using quantitative ISH, we found that AT1a mRNA expression was significantly increased after 5 days of 2% NaCl consumption in anterior third ventricle (AV3V), paraventricular hypothalamus (PVN) and subfornical organ (SFO), but unchanged in anterior pituitary. There were no significant changes in AT1b mRNA. These results document the utility of ISH coupled with quantitative imaging techniques for the study of subtype specific expression. Using ISH and RT-PCR, we verified that AT1a and AT1b receptors are expressed in mouse brain and pituitary and show a similar pattern of distribution. Salt loading produced a specific increase in AT1a mRNA in osmosensitive regions, suggesting that this receptor subtype is regulated by sodium/osmolar input. PMID- 12591118 TI - Hypothalamic cocaine-amphetamine regulated transcript (CART) is regulated by glucocorticoids. AB - Cocaine-amphetamine-regulated transcript (CART) is one of the most abundantly expressed mRNAs in the rat hypothalamus. CART mRNA expression in the arcuate nucleus has been shown to be regulated by leptin, and CART peptides have been implicated in feeding behavior and in the regulation of the HPA-axis. To more fully understand the physiological regulation of CART gene expression, we have examined the effects of adrenalectomy and different types of glucocorticoid substitution (corticosterone and dexamethasone) on hypothalamic CART and POMC mRNA levels. In situ hybridization revealed a reduction in CART mRNA levels in both the hypothalamic paraventricular and arcuate nuclei in adrenalectomized rats, which was fully restored upon dexamethasone treatment but not by a subcutaneous 25% corticosterone pellet. Unlike CART mRNA levels hypothalamic POMC expression was unaltered by adrenenalectomy. The present results show that the CART gene is influenced by glucocorticoids, presumably via a GR dependent mechanism. PMID- 12591119 TI - Neonatal frontal lesion in unilateral hemisphere enhances the development of the intact higher motor cortex in the rat. AB - The influence of the neonatal frontal lesion in unilateral cerebral hemisphere for the organization of intact forelimb motor cortex in the rat was investigated by intracortical microstimulation (ICMS). The relative size of the rostral forelimb area (RFL) compared to the caudal forelimb area (CFL) in the ipsilateral motor field of lesioned rat was significantly greater than those of contralateral in normal and lesioned rats. The optimal sites of the stimulation for ipsilateral responses in lesioned rats were located in the RFL, while the optimal sites for contralateral were located caudolaterally, as for those of normal rats. At the ipsilateral optimal sites within the RFL in the lesioned animals, the threshold for the ipsilateral responses was lower than that for the contralateral responses. That is, the intact hemisphere of the animal preferentially developed the RFL rather than the CFL, for the ipsilateral forelimb. This may suggest a critical role for the RFL in individual forelimb motor control within the normal hemisphere. PMID- 12591120 TI - Proliferation and death of conditionally immortalized neural cells from murine neocortex: p53 alters the ability of neuron-like cells to re-enter the cell cycle. AB - Neurons are distinctive in that they are generally considered to be permanently post-mitotic cells. The oncoprotein p53 is a key regulator in neuronal development, notably in cell proliferation and neuronal death. We hypothesize that p53 maintains the post-mitotic characteristic of differentiated neurons. New lines of conditionally immortalized cortical cells were generated to test this hypothesis. Populations of cells were obtained from the neocortices of dual transgenic mice that were null for p53 and expressed a temperature-sensitive SV40 large T antigen. At a permissive temperature (32 degrees C), the cells continued to proliferate and most expressed nestin and proteins associated with glia. At a non-permissive temperature (39 degrees C), the cells expressed cytoskeletal proteins associated with differentiated neurons such as microtubule associated protein 2 and neurofilament 200. Under permissive conditions, both p53(+/-) and p53(-/-) cells exhibited similar cycling behaviors; the length of the cell cycle was 13-15 h and >85% of the cells were actively cycling. In non-permissive conditions, most p53(+/-) cells stopped dividing, whereas the p53(-/-) cells continued to proliferate. The survival of the cells also differed. In the non permissive conditions, many p53(+/-) cells died following treatment with a neurotoxin (ethanol, 400 mg/dl), whereas the p53(-/-) cells did not. After re introduction to the permissive conditions, both cell lines expressed neuron-like characteristics, but only the p53(-/-) cells retained their ability to cycle. Therefore, p53-mediated activities appear to be involved in the proliferation, survival, and post-mitotic nature of neuron-like cells. PMID- 12591121 TI - Spinal GABAB receptors mediate antinociceptive actions of cholinergic agents in normal and diabetic rats. AB - Spinally administered muscarinic receptor agonists or acetylcholinesterase inhibitors can produce antinociception. However, the mechanisms of the action of cholinergic agents in the spinal cord are not fully understood. Activation of spinal muscarinic receptors evokes gamma-aminobutyric acid (GABA) release, which reduces the glutamatergic synaptic input to dorsal horn neurons through GABA(B) receptors. In this study, we determined the functional role of spinal GABA(B) receptors in the antinociceptive action of intrathecal cholinergic agents in normal rats and in a rat model of diabetic neuropathic pain. Diabetes was induced by intraperitoneal streptozotocin in rats. The intrathecal catheter was inserted with its tip positioned at the lumbar spinal level. Nociceptive threshold was measured by the paw withdrawal latency in response to a radiant heat stimulus in normal rats. Mechanical allodynia in diabetic rats was determined by von Frey filaments applied to the hindpaw. The effect of intrathecal muscarine or neostigmine was examined through pretreatment with the specific GABA(B) receptor antagonist, CGP55845, or its vehicle. Intrathecal injection of muscarine or neostigmine significantly increased the withdrawal latency in response to a heat stimulus in normal rats and the withdrawal threshold in response to application of von Frey filaments in diabetic rats. Intrathecal pretreatment with CGP55845 significantly attenuated the effect of both muscarine or neostigmine in normal rats. Furthermore, the antiallodynic effect of intrathecal neostigmine and muscarine was largely eliminated by CGP55845 in diabetic rats. These data suggest that the GABA(B) receptors in the spinal cord mediate both the antinociceptive and antiallodynic actions of intrathecal muscarine or neostigmine in normal rats and in a rat model of diabetic neuropathic pain. This study provides new functional evidence that activation of spinal GABA(B) receptors is one of the important mechanisms underlying the antinociceptive action of intrathecal cholinergic agents. PMID- 12591122 TI - Heat nociception is severely reduced in a mutant mouse deficient for the L1 adhesion molecule. AB - Recent findings indicate that the spatial organization of the spinal nociceptive reflex system is adjusted postnatally through experience-dependent mechanisms. The cellular and molecular mechanisms underlying this tuning are not known. Because the adhesion molecule L1 is known to play an important role in neural development and synaptic plasticity, we studied the nociceptive withdrawal reflexes in awake adult mutant mice deficient in L1. Withdrawal reflexes were elicited by a CO(2) laser (heat stimulation) and von Frey monofilaments (tactile stimulation). L1-deficient mice (n=10) had an abnormally high nociceptive heat reflex threshold compared with wild-type mice (n=11), except for the nose. Other behavioral signs of heat pain, such as vocalization, were either absent or strongly reduced in L1-deficient mice. Tactile thresholds for withdrawal reflexes were increased in L1-deficient mice when compared with wild-types except for the tail. By contrast, the spatial organization of the withdrawal reflexes appeared normal indicating that the L1 adhesion molecule is not essential for the spatial adjustments of reflex connections during development. The termination patterns of thin primary afferent fibers in the superficial dorsal horn, visualized using intra-plantar injections of WGA-HRP, were normal, suggesting that decreased nociceptive heat sensitivity in L1-deficient mice is mainly due to altered central processing. In view of the known interactions between L1 and some of the NMDA-receptor subtypes, and the prominent role of NMDA receptors in nociception and plasticity, it is conceivable that the hypoalgesia seen in L1 mutants is due, in part, to disturbed NMDA-receptor function. PMID- 12591123 TI - Ultrastructural localization of flotillin-1 to cholesterol-rich membrane microdomains, rafts, in rat brain tissue. AB - There is much interest in research on cholesterol-rich membrane microdomains, rafts, in the field of neurobiology. However, no one has shown the ultrastructure of rafts in tissues. We examined the ultrastructure of rafts in rat brain tissue by pre-embedding immunoelectron microscopy using flotillin-1 antibody, which is a biochemical marker of lipid rafts, and BCtheta, which is nicked and biotinylated theta-toxin, and binds to membrane cholesterol of rafts. Flotillin-1- and BCtheta labeled areas were patchy and prominent on the plasma membranes of small processes and synapses in the neuropil. The size of flotillin-1 labeling was 40 200 nm. In addition, the membrane of lysosome and Golgi apparatus were frequently labeled for flotillin-1 with a patchy pattern. Flotillin-1 and BCtheta were mostly colocalized in double immunolabeling on a part of the plasma membranes of small processes and secondary lysosome membranes. We first indicate that flotillin-1 localizes to BCtheta-positive cholesterol-rich membrane microdomains in vivo, and that flotillin-1 and BCtheta could be ultrastructural raft markers in neural tissue. PMID- 12591125 TI - Intra-accumbens pertussis toxin sensitizes rats to the locomotor activating effects of a single cocaine challenge. AB - Drugs of abuse share common neurochemical signaling substrates, many of which are components of the cAMP cascade. Interestingly, a number of these substrates have been linked to drug-influenced behaviors. This study sought to understand the role of one signaling substrate, inhibitory G-proteins, in a drug-induced phenomenon known as behavioral sensitization. Specifically, we used pertussis toxin (PTX) as a tool to investigate the relationship between cocaine-induced alterations in cAMP signaling and behavior. Vehicle (1 micro l/side) or PTX (0.15 or 0.25 micro g/1 micro l/side) was bilaterally infused into the nucleus accumbens of rats. Locomotor activity was assessed on days 7, 14 and 21 post infusion. Intra-accumbal PTX produced a dose-dependent increase in locomotor activity. On day 21 following behavioral monitoring for 1 h, rats were acutely challenged with cocaine (15 mg/kg, i.p.) and behavioral data were accumulated for an additional 2 h. Intra-accumbal PTX sensitized rats to the locomotor-activating effects of a single cocaine challenge which was dose-dependent. After behavioral testing, brains were removed and processed for in vitro receptor autoradiography using the D(1) receptor ligand [3H] SCH 23390. No changes in D(1) dopamine receptor binding were observed. These findings suggest a role for inhibitory proteins (G(i)/G(o)) within the nucleus acumbens in locomotor activity and also cocaine-induced behavioral sensitization. PMID- 12591124 TI - Rats recovering from unilateral barrel-cortex ischemia are capable of completing a whisker-dependent task using only their affected whiskers. AB - Rats use their vibrissae for a variety of exploratory tasks including location of objects and discrimination of texture. This study examines recovery in vibrissal function following a unilateral ischemic injury to the somatosensory cortex. Vibrissal function was examined in adult food-restricted rats performing on a two texture discrimination device. Animals were trained and tested until the criteria of >80% correct choices was demonstrated on three consecutive days. Ischemic rats were constrained to use the affected whiskers by clipping the ipsilateral vibrissae. One group was tested after ischemia, a second group was trained before ischemia and then tested, and a third group was pre-trained and received whisker stimulation and tested post-ischemia. Nai;ve animals recovering from ischemia took longer to reach criteria than intact or unilateral trimmed control animals. Pre-trained animals with compression ischemia receiving whisker stimulation with sucrose water completed the task to criteria in the fewest number of trials. The results indicate that recovery of vibrissal function occurs following a unilateral ischemic injury. Histological analysis in animals without whisker stimulation indicates that the number of normal appearing cortical barrels following ischemia was inversely correlated to the number of trials needed to complete the behavioral task. This suggests that the natural recovery of the ability to discriminate textures is related to the degree of damage to the barrel cortex. The relationship between cortical barrels and behavioral recovery did not hold for the ischemic animals receiving whisker stimulation. This latter group demonstrated recovery despite marked anatomical lesions suggesting that the intervention influenced reorganization. PMID- 12591126 TI - Dependence on morphine impairs the induction of long-term potentiation in the CA1 region of rat hippocampal slices. AB - The effect of chronic morphine treatment on hippocampal CA1-long-term potentiation (LTP) was examined in vitro. The field excitatory postsynaptic potential (fEPSP) was recorded from stratum radiatum of area CA1 following stimulation of Schaffer collaterals in slices taken from control and morphine dependent rats. To induce LTP, a 100-Hz primed burst stimulation (PBs) was used. Slices from rats exposed to chronic morphine showed no effect on baseline synaptic responses. Slices from control rats or rats exposed to chronic morphine maintained in ACSF with either morphine or naloxone also had no effect on baseline synaptic responses. Control slices perfused with medium containing either morphine or naloxone as well as both drugs exhibited hippocampal CA1 LTP. Similarly, slices from morphine-dependent rats maintained in ACSF with either naloxone or just morphine free ACSF also exhibited hippocampal CA1 LTP. However, slices from morphine-dependent rats maintained in ACSF with morphine significantly attenuated hippocampal CA1 LTP. These findings suggest that hippocampal CA1-LTP can still be achieved in slices from morphine-dependent rats exhibiting morphine withdrawal through mechanisms that may be inhibited by opiate exposure. Such studies can be helpful in understanding the neurophysiological substrate of memory deficits seen in opiate addicts. PMID- 12591127 TI - Interactions of 5-HT2 receptor agonists with acetylcholine in spinal analgesic mechanisms in rats with neuropathic pain. AB - Serotonin type 2 (5-HT(2)) receptors reportedly inhibit neuropathic pain in the spinal cord, but little is known about how spinal 5-HT(2) receptors might act against such abnormal sensitivity. We examined whether the cholinergic and tachykinin systems were involved in the antiallodynic effect of intrathecally administered 5-HT(2) receptor agonists in rats with nerve injury. Allodynia was produced by tight ligation of the left L5 and L6 spinal nerves, and determined by applying von Frey hairs to the left hindpaw. Effects of intrathecal pretreatment with 5-HT(2) receptor antagonists (ketanserin and RS-102221), muscarinic receptor antagonists (atropine and scopolamine), a choline uptake blocker (hemicholium-3), and an NK(1) receptor antagonist (L-706336) were assessed in rats subsequently given a 100- micro g intrathecal dose of a 5-HT(2) receptor agonist either alpha methyl-5-HT or iododimethoxy aminopropane (DOI). Antiallodynic effects of 5-HT(2) receptor agonists were attenuated by the 5-HT(2A) receptor antagonist ketanserin (30 micro g), but not by the 5-HT(2C) receptor antagonist RS-102221 (40 micro g). Muscarinic receptor antagonists (30 micro g each), the choline uptake blocker (10 micro g), and the NK(1) receptor antagonist (30 micro g) also inhibited the antiallodynic effects of 5-HT(2) receptor agonists. Antiallodynic effects of intrathecally administered 5-HT(2) receptor agonists may be mediated by spinal release of acetylcholine induced via 5-HT(2A) and NK(1) receptors. PMID- 12591128 TI - Induction of neuronal nitric oxide synthase by sympathetic denervation is mediated via alpha 2-adrenoceptors in the jejunal myenteric plexus. AB - Nitric oxide (NO) is an important nonadrenergic, non-cholinergic (NANC) inhibitory neurotransmitter in the gastrointestinal tract. In previous studies, neuronal nitric oxide synthase (nNOS) in the jejunal myenteric plexus, a key enzyme responsible for the release of NO, has been demonstrated to increase after splanchinic ganglionectomy (sympathetic nerve transection). The alpha2 adrenoceptor is known to be one of the most important receptors which controls intestinal motility. In the present study, we examined the effect of application of the alpha2-adrenoceptor agonist, clonidine hydrochloride, on nNOS expression in the rat jejunal myenteric plexus after splanchinic ganglionectomy. Clonidine (0.1-1 mg/kg, i.p.) or saline was administered for 5 days after the splanchinic ganglionectomy. The nNOS expression and nNOS mRNA were detected by immunohistochemistry and in situ hybridization for nNOS mRNA, respectively. In the rats treated with vehicle after the splanchinic ganglionectomy, nNOS expression in the myenteric plexus significantly increased compared with sham operated rats. The increases in nNOS protein and mRNA after splanchinic ganglionectomy were significantly reversed by clonidine treatment. Clonidine treated naive rats showed no difference in nNOS expression compared with sham operated rats. These data suggest that nNOS expression in the jejunal myenteric plexus after splanchinic ganglionectomy is regulated by the alpha2-adrenoceptor and that the alpha2-adrenoceptor may play an important role in abnormal intestinal motility following splanchinic ganglionectomy in rat jejunum. PMID- 12591129 TI - Neuroprotective effects of antioxidative flavonoids, quercetin, (+) dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. saboten. AB - The flavonoids quercetin, (+)-dihydroquercetin, and quercetin 3-methyl ether were isolated from the ethyl acetate fractions of the fruits and stems of Opuntia ficus-indica var. saboten. In the present study, we evaluated their protective effects against oxidative neuronal injuries induced in primary cultured rat cortical cells and their antioxidant activities by using three different cell free bioassays. Quercetin was found to inhibit H(2)O(2)- or xanthine (X)/xanthine oxidase (XO)-induced oxidative neuronal cell injury, with an estimated IC(50) of 4-5 micro g/ml. However, it was no more protective at concentrations of 30 micro g/ml and above. (+)-Dihydroquercetin concentration-dependently inhibited oxidative neuronal injuries, but it was less potent than quercetin. On the other hand, quercetin 3-methyl ether potently and dramatically inhibited H(2)O(2)- and X/XO-induced neuronal injuries, with IC(50) values of 0.6 and 0.7 micro g/ml, respectively. All three principles markedly inhibited lipid peroxidation and scavenged 1,1-diphenyl-2-picrylhydrazyl free radicals. In addition, quercetin and quercetin 3-methyl ether were shown to inhibit XO activity in vitro, with respective IC(50) values of 10.67 and 42.01 micro g/ml. These results indicate that quercetin, (+)-dihydroquercetin, and quercetin 3-methyl ether are the active antioxidant principles in the fruits and stems of Opuntia ficus-indica var. saboten exhibiting neuroprotective actions against the oxidative injuries induced in cortical cell cultures. Furthermore, quercetin 3-methyl ether appears to be the most potent neuroprotectant of the three flavonoids isolated from this plant. PMID- 12591130 TI - Neuroprotective action of tacrolimus (FK506) in focal and global cerebral ischemia in rodents: dose dependency, therapeutic time window and long-term efficacy. AB - Tacrolimus (FK506), a potent immunosuppressive drug, is effective in attenuating brain infarction after cerebral ischemia. However, there has been no report characterizing the neuroprotective action and therapeutic time window of tacrolimus systematically using different types of stroke models and extended observation periods. Therefore, we evaluated the neuroprotective effect of tacrolimus in three different animal models of cerebral ischemia: transient and permanent focal ischemia in rats and transient global ischemia in gerbils. Tacrolimus at doses higher than 0.1 mg/kg (i.v.) produced a statistically significant reduction in ischemic brain damage following permanent and transient focal ischemia in rats when administered immediately after the onset of ischemia. Tacrolimus (1 mg/kg, i.v.) demonstrated similar neuroprotective activity even after delayed administration (2 h after permanent or 1 h after transient focal ischemia). The neuroprotective effect of tacrolimus was still present 2 weeks after transient focal ischemia and 1 week after permanent focal ischemia. After transient global ischemia in gerbils, tacrolimus (1 mg/kg, i.v.) given immediately after reperfusion also produced long-lasting neuroprotective effects with a protective time-window of 1-2 h. Taken together, the results clearly indicate that tacrolimus exerts potent, long-term neuroprotective effects with a favorable therapeutic time-window, regardless of the model of cerebral ischemia. These results strengthen the notion that tacrolimus might be of clinical value for the treatment of acute stroke. PMID- 12591131 TI - Firing of micturition center neurons in the rat mesopontine tegmentum during urinary bladder contraction. AB - Micturition is controlled by a network of brainstem neurons involving the Barrington's nucleus. To depict clearly the brainstem system for micturition control, the present study was designed to record single neuronal activity in the mesopontine tegmentum including the Barrington's nucleus, and to observe its precise timing in relation to bladder contraction recorded simultaneously. About 1/5 of neurons encountered had firing modulated in relation to bladder contraction. Three types of neurons were distinguished; those which fired only prior to the start of contraction (type E1), those whose firing started shortly prior to and was maintained during contraction (type E2), and those whose firing was strongly suppressed during contraction (type I). Type E2 neurons were most frequently observed in the Barrington's nucleus and its close vicinity, while the neurons of the other two types were scattered widely in the mesopontine tegmentum. The results show clearly that direct neural signals to induce bladder contraction may arise from the Barrington's nucleus, and that the nucleus may receive regulatory inputs from wide areas of the mesopontine tegmentum. In addition, the present study clarified that the noradrenergic and cholinergic neurons, which are located in nuclei adjoining the Barrington's nucleus and function to control sleep/wakefulness, may not be concerned in controlling micturition directly. PMID- 12591132 TI - Effect of new O-superfamily conotoxin SO3 on sodium and potassium currents of cultured hippocampal neurons. AB - The effects of a new O-superfamily conotoxin SO3 on sodium and potassium currents were examined in cultured rat hippocampal neurons using the whole-cell patch clamp technique. SO3 caused a concentration-dependent, rapidly developing and reversible inhibition of sodium currents (I(Na)). The IC(50) value for the blockage of I(Na) was calculated to be 0.49 and the Hill coefficient was 1.7. Using electrophysiological and pharmacological protocols, transient A-type potassium currents (I(A)) and delayed rectifiers potassium currents (I(K)) were isolated. SO3 caused a concentration-dependent, and reversible inhibition of I(K). The IC(50) value for the blockage of I(K) was calculated to be 1.6 and the Hill coefficient was 0.6, with no significant effect on I(A). These results indicate that SO3 can selectively inhibit neuronal sodium and potassium currents. PMID- 12591133 TI - Topographical anatomy of the cerebellum in the guinea pig, Cavia porcellus. AB - Zebrin II/aldolase C is expressed in a stereotyped array of parasagittal bands and transverse zones in the cerebellum of many animals including birds and mammals. Here, section and whole mount immunohistochemistry has been used to characterize the expression of zebrin II in the cerebellum of the adult guinea pig. Purkinje cells in the adult guinea pig express zebrin II immunoreactivity at three different levels of intensity-high, medium and low. This expression pattern reveals an arrangement of parasagittal bands that are symmetrical about the midline and reproducible between individuals. The expression of zebrin II divides the vermis into four transverse expression domains from rostral to caudal: an anterior zone consisting of one zebrin II-immunoreactive band at the midline and at least three symmetrical bands laterally; a central zone, in which broad zebrin II-positive bands are separated by narrow bands of zebrin II-negative Purkinje cells that disappear caudally to leave no overt compartmentation; a posterior zone consisting of alternating bands of zebrin II-positive and -negative Purkinje cells; and finally, a nodular zone in which nearly all Purkinje cells express zebrin II. In the anterior and posterior hemispheres, zebrin II is also expressed in a banded pattern. These rostrocaudal and mediolateral patterns of zebrin II expression are reminiscent of those in other mammals including rabbit, rat, and mouse, and suggest that there may be a fundamental compartmental organization of the cerebellum that is conserved in mammals. PMID- 12591134 TI - Alteration of zinc concentrations in the brain implanted with C6 glioma. AB - The uptake of zinc, an essential element, is critical not only for cell proliferation but also cell functions. On the basis of the evidence suggesting that preferential uptake of 65Zn in brain tumors may be associated with avid cell proliferation, alteration of zinc concentrations in the brain implanted with C6 glioma was studied by brain autoradiography using 65Zn. After implantation of C6 glioma cells into the hippocampus, 65Zn concentrations in the brain were compared between 1 h and 6 days after intravenous injection of 65ZnCl(2). The 65Zn concentration in the tumor 1 h after the injection was approximately twice as high as that 6 days after the injection. However, the 65Zn concentration in the tumor 6 days after the injection was higher than in any other brain region. These results suggest that zinc taken up in the tumor may be utilized for tumor cell proliferation and that there is greater potential in the use of 69mZn, a short half life gamma-emitter, for evaluation of the viability of brain tumors. PMID- 12591135 TI - Decreased limbic vesicular monoamine transporter 2 in a genetic rat model of depression. AB - Psychopharmacological and neurochemical research suggests that alterations in monoamine transporters may be involved in the etiology of depression. We studied the expression of the brain-type vesicular monoamine transporter (VMAT2) in the Flinders sensitive line (FSL) rats, which represent a genetic animal model for clinical depression in humans. VMAT2 expression at the protein level was assessed by autoradiography using [(3)H]dihydrotetrabenazine ([(3)H]TBZOH) binding, in the prefrontal cortex, the striatum and its subregions, nucleus accumbens (NAC), ventral tegmental area (VTA) and the substantia nigra pars compacta (SNC). The VMAT2 mRNA level was analyzed by in situ hybridization, in the VTA, SNC and the dorsal raphe (DR) nucleus. Reduced levels of [(3)H]TBZOH binding were detected in the striatum and its subregions, NAC shell but not in the NAC core. A marked reduction of 21% was found in the VTA while only a slight reduction (13%) was observed in the SNC. The reduced levels of VMAT2 protein binding capacity were not accompanied by a parallel alteration in VMAT2 mRNA levels in the VTA, the SNC and the DR. Since the VMAT2 is responsible for the intracellular storage and regulated release of monoamines, the reduced [(3)H]TBZOH binding levels in limbic brain regions of FSL rats may imply a reduced density of vesicular monoamine transporters, which can result in reduced monoamine transmission. Such reduction in the limbic neurotransmission, especially in NAC shell and VTA regions, may be involved in the depressive features of anhedonia and lack of motivation reported in the FSL rats. PMID- 12591136 TI - The cholinesterase inhibitor DFP facilitates the expression of paradoxical sleep (PS) propensity in rats subjected to short-term PS deprivation. AB - Short-term paradoxical sleep (PS) deprivation was used to examine the effects of chronic exposure to subtoxic doses of the cholinesterase inhibitor diisopropylfluorophosphate (DFP) on PS regulation. Rats were injected once daily with DFP (0.2 mg/kg per day; s.c.) for 11 consecutive days; control rats received a daily injection of oil vehicle. The experiment was conducted on the 10th and 11th days of treatment, when brain cholinesterase inhibition induced by DFP exposure was maximal. On the 10th day, an 8-h baseline recording was carried out. On the 11th day, a 6-h PS deprivation was carried out by manually awaking rats each time they showed polygraphic signs of PS; recordings were then continued for another 2 h to examine recovery sleep. During deprivation, though they slept less than controls, DFP-treated rats made more attempts to enter PS. After deprivation, their PS rebound had an overall amount comparable to that of the controls, but its time course was shortened: whereas PS elevation was manifested through the 2 h of recovery in the control group, it occurred only during the first hour in the DFP group. These results demonstrate that chronic, low-level DFP exposure facilitated the expression of the PS propensity that accumulated as a result of PS deprivation: it enhanced the tendency for PS during deprivation; it accelerated the rate of compensatory PS expression after deprivation. They support the hypothesis that DFP promotes PS initiation by increasing cholinergic transmission. PMID- 12591137 TI - Distinct neurochemical mechanisms are activated following administration of different P2X receptor agonists into the hindpaw of a rat. AB - Nocifensive behaviors induced by the intradermal injection of three different P2X receptor agonists, ATP, BzATP or alpha,beta-meATP, into a hindpaw were measured in rats that were injected intrathecally with either an NMDA (MK-801) or an NK-1 (L-703,606) receptor antagonist or were pretreated systemically with the VR1 agonist resiniferatoxin (RTX). The same procedures were performed in animals injected intradermally with either capsaicin or formalin. Spinal infusion of MK 801 (10-50 nmol/10 micro l) similarly reduced the number of nociceptive events triggered by each of the P2X agonists and was also effective against capsaicin and formalin induced behaviors. Intrathecal administration of L-703,606 (50-100 nmol/10 micro l) had its greatest antinociceptive effect against capsaicin induced behaviors followed by ATP and BzATP. L-703,606 was completely ineffective against behaviors induced by formalin or the other P2X agonist, alpha,beta-meATP. Pretreatment with RTX 2 days prior to testing significantly decreased the number of nociceptive events caused by each of the P2X agonists as well as capsaicin and formalin (capsaicin>BzATP>ATP>formalin>alpha,beta-meATP). The remaining nociceptive events in RTX animals injected with alpha,beta-meATP were significantly higher than in animals injected with either ATP or BzATP. Intradermal administration of different P2X receptor agonists induced similar levels of nocifensive behaviors and activity at spinal NMDA receptors. Capsaicin sensitive fibers were likely activated following injection of BzATP and ATP, but not alpha,beta-meATP, and appeared to trigger the spinal release of substance P. The differences in mechanisms employed by the different P2X agonists may be a function of respective selectivity for P2X receptor subtypes. PMID- 12591138 TI - Specific binding of 125I-hippocampal cholinergic neurostimulating peptide (HCNP) to rat brain membranes: characterization and regional distribution. AB - An undecapeptide-hippocampal cholinergic neurostimulating peptide (HCNP), originally purified from young rat hippocampus, enhances cholinergic phenotype development in the medial septal nucleus in vitro. To survey and characterize the HCNP receptor within the central nervous system, we used iodinated HCNP as a labeled ligand. In preliminary experiments, [125I]HCNP binding was highest in the crude P2 membrane fraction, so all subsequent experiments were performed using this fraction. The binding was saturable and reversible, and unlabeled ligand inhibited it. Scatchard analysis of the concentration-dependent saturation of binding indicated a single population of non-interacting sites with K(d) 4.0+/ 0.7 nM and B(max) 10.7+/-3.8 pmol/mg protein. Dissociation experiments revealed a dissociation constant (K(-1)) of 0.07 min(-1). Inhibition experiments using HCNP and its shorter peptide fragments suggested that the active binding site resided close to the peptide's C-terminal sequence. Since [125I]HCNP binding was found in crude P2 membrane fractions from animals at all ages examined, HCNP may also perform important functional roles in the adult brain. Further, the predominant distribution of the receptor in the P2 membrane fraction, and the similarity in distribution patterns between the binding site and HCNP-precursor protein mRNA expression suggest that the peptide exerts its functions in the vicinity of the dendrites of the neurons that produce it. PMID- 12591139 TI - Cohabitation induced Fos immunoreactivity in the monogamous prairie vole. AB - Cohabitation of sexually nai;ve male and female prairie voles (Microtus ochrogaster) triggers a cascade of physiological changes that result in the formation of stable pair bonds. In the present study we used the expression of c Fos protein to identify CNS regions activated during initial social contact in heterosexual, male/male and female/female pairs. Sexually naive males and females were randomly assigned to one of five groups: control- no cohabitation, or cohabitation for 1 h with an unrelated opposite sex, an unrelated same sex, an unfamiliar same sex sibling, or removal for 24 h and then repaired with the familiar sibling. Heterosexual pairing resulted in significant increases in c-Fos immunoreactivity (IR) in the posterodorsal and posteroventral medial amygdala (MeA), bed nucleus of the stria terminalis, medial preoptic nucleus, ventrolateral portion of the ventromedial nucleus of the hypothalamus (VMN-VL) in males and females, and the periventricular nucleus of the thalamus in males only. c-Fos IR during intrasexual cohabitation varied with the relationship of the experimental animal to the stimulus animal. Males cohabited with an unfamiliar unrelated male expressed significantly more c-Fos IR in the central amygdala (CeA). While females cohabited with an unfamiliar female (related or unrelated) also displayed increased c-Fos IR in the CeA, this increase was accompanied by an increase in c-Fos IR in the VMN-VL and MeA. The results from this study suggest that early neuronal activation associated with heterosexual cohabitation is similar in both sexes, while neuronal activation is sexually dimorphic in response to intrasexual cohabitation. PMID- 12591140 TI - Involvement of dopamine D1 receptors of the central amygdala on the acquisition and expression of morphine-induced place preference in rat. AB - In the present study, the effects of intra-central amygdala (CeA) injection of dopamine D1 receptor agonist and antagonist on morphine-induced conditioned place preference (CPP) were investigated in male Wistar rats. Our data showed that subcutaneous (s.c.) injection of morphine sulphate (0.5-10 mg/kg) significantly increased the time spent in the drug-paired compartment in a dose-dependent manner. Intra-CeA administration of the dopamine D1 receptor agonist, SKF 38393 (2 and 4 micro g/rat) with an ineffective dose of morphine (0.5 mg/kg), elicited a significant conditioned place preference. On the other hand, a single dose of SKF 38393 (2 micro g/rat, intra-CeA) in combination with the lower doses (0.5 and 2.5 mg/kg), but not with the higher doses of morphine potentiated morphine induced CPP. Furthermore, intra-CeA administration of the dopamine D1 receptor antagonist, SCH 23390 (0.5-1 micro g/rat) decreased the acquisition of conditioned place preference induced by morphine (7.5 mg/kg). The response of SKF 38393 was decreased by SCH 23390 (0.75 micro g/rat). SKF 38393 or SCH 23390 by themselves did not elicit any effect on place conditioning. On the other hand, intra-CeA administration of SKF 38393 or SCH 23390 significantly decreased the expression of morphine (7.5 mg/kg)-induced place preference. SKF 38393 or SCH 23390 injections into the CeA had no effects on the locomotor activity on the test sessions. The results indicate that the dopamine D1 receptors in the CeA may be involved in the acquisition and expression of morphine-induced place preference. PMID- 12591141 TI - Effect of cortical masticatory area stimulation on swallowing in anesthetized rabbits. AB - The effects of stimulation of the cortical masticatory area (CMA) on swallowing evoked by superior laryngeal nerve (SLN) were studied in anesthetized rabbits. Electromyographic activity of the thyrohyoid, masseter, and digastric muscles and jaw-movement trajectories were recorded to monitor rhythmic jaw movements (RJMs) or swallowing. A systematic series of microelectrode penetrations within the CMA was made for each animal, and the effects of CMA stimulation on swallowing were tested by comparing the number of swallows evoked by stimulation of the CMA alone, the SLN alone, and simultaneous stimulation of the SLN and CMA. A significant facilitatory effect was observed in 49 (52%) of the 95 CMA loci tested. No significant effect was noted in the remaining 46 loci. Three different types of RJMs were evoked by CMA stimulation, and topographical organization was noted among CMA loci that evoked different types of RJMs. A high percentage of (77%) the CMA loci that evoked RJMs with a prominent horizontal excursion of the jaw facilitated swallowing and was located in the posterolateral and deep part of the CMA. A majority (88%) of the CMA loci that evoked RJMs with small circular jaw movements did not affect swallowing and was located in the anteromedial and shallow part of the CMA. The facilitatory effect of CMA stimulation on swallowing remained even after removal of peripheral sensory inputs by means of deafferentation of infraorbital and inferior alveolar nerves. Results suggest the existence of facilitatory descending pathways to the swallowing center from particular intracortical loci of CMA. PMID- 12591142 TI - Novel function of bovine milk-derived lactoferrin on antinociception mediated by mu-opioid receptor in the rat spinal cord. AB - Lactoferrin (LF) is a multifunctional protein that is found in milk, neutrophils, and other biological fluids. Although LF and the LF receptor have been identified in the central nervous system (CNS), the physiological role of LF remains unknown. We found that bovine milk-derived LF (BLF) reduces nociception in various pain models, as shown by the formalin test, hot plate test, and acetic acid writhing test in rats. Intraperitoneal (i.p.) administration of BLF significantly inhibited nociception in these pain models. These antinociceptive effects were also confirmed in BLF-fed rats. The antinociceptive effects of BLF were blocked by naloxone treatment, even though prostaglandin E(2) (PGE(2)) production in the ascites fluid that accumulated during the writhing test was not affected by BLF. Intrathecal (i.t.) application of BLF caused marked antinociceptive effects that were reversed by co-administration of a specific mu opioid receptor antagonist, D-Phe-Cys-Tyr-D-Trp-Orn-Thr-NH(2) (CTOP), or by naloxone during the formalin test. We conclude that LF possesses mu-opioid receptor-mediated antinociceptive activity in the spinal cord. PMID- 12591143 TI - Somatostatin-like immunoreactivity in the brain of the urodele amphibian Pleurodeles waltl. Colocalization with catecholamines and nitric oxide. AB - The neuronal structures with somatostatin-like immunoreactivity have been studied in the brain of the urodele amphibian Pleurodeles waltl. Intense immunoreactivity was observed in neurons and fibers distributed throughout the brain. Within the telencephalon, the subpallial regions were densely labeled containing both cells and fibers, primarily in the striatum and amygdala. The majority of the somatostatin immunoreactive neurons were located in the preoptic area and hypothalamus, although less numerous cells were also found in the thalamus. A conspicuous innervation of the median eminence was revealed, which arises from the hypothalamic cell populations. In the brainstem, intense fiber labeling was present in the tectum and tegmentum, whereas cell bodies were located only in the tegmentum of the mesencephalon and in the interpeduncular, raphe and reticular nuclei of the rhombencephalon. Longitudinal fiber tracts throughout the brainstem were observed and they continued into the spinal cord in the laterodorsal funiculus. The localization of somatostatin in catecholaminergic and nitrergic neurons was studied by double labeling techniques with antisera against tyrosine hydroxylase and nitric oxide synthase. Catecholamines and somatostatin only colocalized in a cell population in the ventral preoptic area. In turn, the striatum and amygdala contained neurons with somatostatin and nitric oxide synthase. Our results demonstrated that the somatostatin neuronal system in the brain of Pleurodeles waltl is consistent with that observed in anuran amphibians and shares many characteristics with those of amniotes. Colocalization of somatostatin with catecholamines and nitric oxide is very restricted in the urodele brain, but in places that can be easily compared to those reported for mammals, suggesting that interactions between these neurotransmitter systems are a primitive feature shared by tetrapod vertebrates. PMID- 12591144 TI - Repeated swim stress increases pain-induced expression of c-Fos in the rat lumbar cord. AB - We have previously demonstrated that repeated swim stress produces a long-lasting cutaneous hyperalgesia in rats. We have now looked at c-Fos expression in the spinal lumbar cord of male Sprague-Dawley rats subjected to 10-20 min daily sessions of forced swimming for 3 consecutive days. Control rats were subjected to sham swimming or were completely naive. Forty-eight hours later, nociception was assessed by recording for 90 min the nociceptive behavior evoked the injection of 1% formalin in the hind paw. Thirty min later, the rats' spinal cords were removed for c-Fos immunocytochemistry. Total pain scores were 45% higher in swim stressed rats compared to control animals due an increased nociceptive behavior during last 70 min of the recording period. In addition, the number of c-Fos-immunoreactive nuclei was 40% higher in the lumbar ipsilateral dorsal horn (L4-L5) of swim stressed rats than in controls, being the highest relative increase, relative to the control groups, observed in laminae III-IV, followed by laminae V-VI, with the smallest difference in laminae I-II. c-Fos expression in the contralateral dorsal horn was higher in swim stressed rats than in sham and nai;ve rats. In the absence of a nociceptive stimulus, a low level of c-Fos expression was observed mainly in laminae I, II, V, and VI, being higher in swim stressed rats than in sham rats. These findings suggest that repeated inescapable and uncontrollable stress could induce a sensitization and activation of sensory neurons at the spinal level. PMID- 12591145 TI - Axon terminals possessing the alpha 2c-adrenergic receptor in the rat dorsal horn are predominantly excitatory. AB - In this study we used confocal microscopy to show that most (83.67%) alpha(2C) adrenergic receptor (alpha(2C)-AR)-immunoreactive terminals in the superficial dorsal horn contain the vesicular glutamate transporter 2 and hence are glutamatergic. Few (11.33%) terminals contain glutamic acid decarboxylase (a marker for GABAergic axons) and none were associated with the B subunit of cholera toxin (a marker for myelinated primary afferents) or the vesicular glutamate transporter 1. These data indicate that most dorsal horn axons possessing the alpha(2C)-AR are excitatory and add further support to the suggestion that they originate principally from spinal interneurons. PMID- 12591146 TI - Evidence in vitro of glial cell priming in the taiep rat. AB - Cultured glial cells from the cerebellum of 15-day-old taiep rats produced NO, increased iNOS levels, up-regulated iNOS expression and promoted TNF release when stimulated with LPS and IFNgamma. These responses were much greater than in control cells. In taiep glial cells, NO production and iNOS levels and expression induced by the co-stimulatory signal were resistant to the inhibitory effect of TGFbeta1. The glial cell priming might have been generated by oligodendrocyte alteration in taiep rats. PMID- 12591147 TI - Influence of manganese on the release of neurotransmitters in rat striatum. AB - On the basis of the evidence that manganese may be released along with glutamate into the extracellular space in the hippocampus and amygdala, the release of manganese and its influence in the striatum was examined by using the in vivo microdialysis method in the present study. The release of 54Mn previously taken up by the striatum into the extracellular space was enhanced during stimulation with 100 mM KCl. This enhancement of 54Mn release into the striatal extracellular space was inhibited by addition of 1 micro M tetrodotoxin. When the rat striatum was perfused with artificial CSF containing 200 nM MnCl(2), the levels of GABA in the perfusate were remarkably decreased, while the levels of glutamate, aspartate, and glycine in the perfusate were not appreciably decreased. These results suggest that manganese released into the synaptic cleft in a calcium- and impulse-dependent manner may influence GABA release in the striatum. PMID- 12591148 TI - Acetylcholinesterase is increased in mouse neuronal and astrocyte cultures after treatment with beta-amyloid peptides. AB - The cellular origin of the acetylcholinesterase (AChE) associated with amyloid plaques in the Alzheimer's disease (AD) brain is unknown. In this study we report that amyloid beta-peptides (Abeta) increased AChE levels in both neuronal and astrocytic primary cultures, supporting the possibility that both neurons and glia may make a direct contribution to the pool of AChE seen around amyloid deposits in the AD brain. PMID- 12591149 TI - Calcium overload enhances hydroxyl radical generation by 1-methyl-4 phenylpyridinium ion (MPP+) in rat striatum. AB - We examined whether ouabain-induced Ca(2+) overload increases hydroxyl radical (*OH) generation by 1-methyl-4-phenylpyridinium ion (MPP(+)) in rat striatum. These elevations seem to induce lipid peroxidation of striatum of rats, as detected by increases in non-enzymatic formation of 2,3-dihydroxybenzoic acid (DHBA) levels. Ouabain enhanced MPP(+)-induced *OH formation trapped as DHBA. Moreover, when iron (II) was administered to MPP(+) then ouabain (100 micro M) pretreated animals, a marked elevation in the level of DHBA was observed, as compared with the iron (II)-only-treated animals. These results suggests that Ca(2+) overload might enhance *OH generation by MPP(+) in rat striatum. PMID- 12591150 TI - Effects of chronic stress on dendritic arborization in the central and extended amygdala. AB - A differential role has been suggested for two important areas in the neural circuitry of stress, central nucleus of the amygdala (CeA) and bed nucleus of stria terminalis (BNST) in the extended amygdala, in regulating fear versus anxiety. Since chronic stress enhances anxiety and consolidation of aversive memories, we examined the effects of chronic immobilization stress (CIS) on neuronal morphology in the CeA and BNST of rats. In contrast to previous reports of stress-induced atrophy in the hippocampus, CIS does not cause dendritic atrophy in CeA and BNST neurons. While dendritic arborization in CeA neurons remains unaffected, it increases in BNST neurons after CIS. These results suggest a role for dendritic remodeling of BNST neurons in stress-induced facilitation of anxiety. PMID- 12591151 TI - Dose-dependent effect of cholinotoxin AF64A on the cholinergic elements of the cingulum bundle in rat. AB - Previous studies revealed that cholinergic neurons possessing long axons are extremely sensitive to ethylcholine aziridinium ion (AF64A) administration [Neuropharmacology 31 (1992) 397]. In the present paper we examined the effect of AF64A on the cholinergic elements of the cingulum bundle. Seven days after AF64A administration choline acetyltransferase (ChAT)-immunoreactive fibers were extensively damaged on the dorsal part of cingulum bundle. These findings are the first reporting damage by AF64A to this brain region. PMID- 12591152 TI - Retraction. Kynurenate attenuates the accumulation of diacylglycerol and free fatty acids after experimental brain injury in the rat. [Brain Research 832 (1999) 7-12]. PMID- 12591153 TI - Transfer of NMDAR2 cDNAs increases endogenous NMDAR1 protein and induces expression of functional NMDA receptors in PC12 cells. AB - The pheochromocytoma cell line (PC12) has been used as a model system for the study of regulation of expression of NMDA receptors. PC12 cells express a substantial amount of NMDAR1 subunit (NR1) mRNA, whereas they express only a small amount of NR1 protein. The level of functional NMDA receptor expression is almost negligible. To test the possibility that NMDAR2 subunits (NR2) control expression of functional NMDA receptors as well as NR1 protein, we transferred NR2A-D cDNAs into PC12 cells using adenovirus vectors. Prominent NMDA receptor mediated currents were recorded in PC12 cells to which NR2A or NR2B cDNA was delivered without NR1 cDNA. The amplitudes of these responses were similar to those in PC12 cells to which NR1 cDNA was delivered together with NR2A or NR2B cDNA. In cells to which either NR2C or NR2D cDNA alone was delivered, NMDA receptor-mediated currents were also detected, although to a much lesser extent. These results showed that NR2 proteins produced by gene transfer are co-assembled with the endogenous NR1 protein to form functional heteromeric receptors. The delivery of NR2A-D cDNAs also increased the amount of NR1 protein but not that of NR1 mRNA, suggesting that this protein increase is due to post-transcriptional mechanisms. The effects of NR2A-B gene transfer on expression of NR1 protein were much more efficient than those of NR2C-D gene transfer. PMID- 12591154 TI - Increased neuropeptide Y mRNA expression in striatum in Parkinson's disease. AB - High levels of neuropeptide Y (NPY) are found in basal ganglia where it is co localised with somatostatin (SOM) and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH/d) in a population of striatal GABA containing interneurones. Although alterations occur in the levels of various neuropeptides in basal ganglia in Parkinson's disease (PD), it is not known whether NPY is affected. Using in situ hybridisation immunohistochemistry, we have examined the distribution of NPY mRNA in the caudate nucleus, putamen and nucleus accumbens of normal individuals and patients with PD. NPY mRNA was weakly expressed in the caudate nucleus, putamen and nucleus accumbens in normal individuals with a scattered labelling of neurones. However, there was no regional localisation within any brain area and no obvious differences between brain regions. In PD, the number of NPY mRNA-expressing cells was increased as was the density of the silver grains overlying each positive cell. The increase was more pronounced in the nucleus accumbens and in the ventral part of the caudate nucleus. The increase in NPY mRNA expression observed in patients with PD may reflect the loss of dopaminergic tone on striatal NPY containing interneurones, although a role for chronic L-DOPA therapy cannot be ruled out. PMID- 12591155 TI - Reissner fiber binds and transports away monoamines present in the cerebrospinal fluid. AB - The subcommissural organ (SCO) is a brain gland that secretes glycoproteins into the cerebrospinal fluid (CSF), where they subsequently aggregate to form Reissner fiber (RF). By addition of newly released glycoproteins to its cephalic end, RF grows constantly through the Sylvian aqueduct, fourth ventricle and central canal of the spinal cord. Disaggregation of RF-material and passage to blood occur when RF reaches the terminal ventricle at the filum. The present investigation was designed to test the hypothesis that RF binds, transports and clears away monoamines present in the CSF. Four experimental protocols were applied: (i) in vivo binding of labeled monoamines to the rat RF, studied by pulse and chase, and after perfusion for 7 days; (ii) identification of monoamines, by high performance liquid chromatography (HPLC), naturally occurring in the bovine RF; (iii) in vitro binding of labeled and unlabeled monoamines to the isolated bovine RF; and (iv) tentative identification of the amine binding site(s) in RF-proteins by use of specific antibodies. The results obtained indicate that RF participates in the regulation of the CSF concentration of monoamines either by binding and transporting them away throughout the central canal of the spinal cord (L-DOPA, noradrenaline, adrenaline), or by transiently binding them and releasing them back to the CSF (serotonin). Furthermore, evidence was obtained that (i) adrenaline and noradrenaline share the same binding site, and that this site would correspond to a repeated sequence present in the SCO-spondin, the major protein component of RF; and (ii) serotonin has its own binding site in RF. PMID- 12591156 TI - Age-related working memory impairment is correlated with increases in the L-type calcium channel protein alpha1D (Cav1.3) in area CA1 of the hippocampus and both are ameliorated by chronic nimodipine treatment. AB - The hippocampus is critical for spatial memory formation in rodents. Calcium currents through L-type voltage-sensitive calcium channels (L-VSCCs) are increased in CA1 neurons of the hippocampus of aged rats. We have recently shown that expression of the calcium conducting L-VSCC subunit alpha(1D) (Ca(v)1.3) is selectively increased in area CA1 of aged rats. We and others have speculated that excessive Ca(2+) influx through L-VSCC may be detrimental to memory formation. Therefore, we investigated the relationship between age-related working memory decline and alpha(1D) protein expression in the hippocampus. In addition, we studied the effects of chronic treatment with the L-VSCC antagonist nimodipine (NIM) on age-related working memory deficits and alpha(1D) expression in the hippocampus. Here we report that age-related increases in alpha(1D) expression in area CA1 correlate with working memory impairment in Fischer 344 rats. Furthermore, we demonstrate that chronic NIM treatment ameliorates age related working memory deficits and reduces expression of alpha(1D) protein in the hippocampus. The present results suggest that L-VSCCs participate in processes underlying memory formation and that increases in L-VSCC protein and currents observed with aging may play a role in age-related memory decline. Furthermore, the amelioration in age-related memory decline produced by NIM treatment may be mediated, at least in part, by reductions in the abnormally high levels of alpha(1D) protein in the aged hippocampus. These findings may have implications for patients with Alzheimer's disease, who show increased L-VSCC protein expression in the hippocampus, and for patients receiving chronic treatment with L-VSCC antagonists. PMID- 12591157 TI - Kir2 potassium channels in rat striatum are strategically localized to control basal ganglia function. AB - Parkinson's disease is the most frequent movement disorder caused by loss of dopaminergic neurons in the midbrain. Intentions to avoid side effects of the conventional therapy should aim to identify additional targets for potential pharmacological intervention. In principle, every step of a signal transduction cascade such as presynaptic transmitter release, type and occupation of postsynaptic receptors, G protein-mediated effector mechanisms, and the alterations of pre- or postsynaptic potentials as determined by the local ion channel composition, have to be considered. Due to their diversity and their widespread but distinct localizations, potassium channels represent interesting candidates for new therapeutic strategies. As a first step, the present report aimed to study in the striatum the cellular and subcellular distribution of the individual members of the Kir2 family, a group of proteins forming inwardly rectifying potassium channels. For this purpose polyclonal monospecific affinity purified antibodies against the less conserved carboxyterminal sequences from the Kir2.1, Kir2.2, Kir2.3, and Kir2.4 proteins were prepared. All subunits of the Kir2 family were detected on somata and dendrites of most striatal neurons. However, the distribution of two of them was not homogeneous. Striatal patch areas were largely devoid of the Kir2.3 protein, and the Kir2.4 subunit was most prominently expressed on the tonically active, giant cholinergic interneurons of the striatum. These two structures are among the key players in regulating dopaminergic and cholinergic neurotransmission within the striatum, and therefore are of major importance for the output of the basal ganglia. The heterogeneous localization of the Kir2.3 and the Kir2.4 subunits with respect to these strategic structures pinpoints to these channel proteins as promising targets for future pharmacological efforts. PMID- 12591158 TI - Double lox targeting for neural cell transgenesis. AB - ES cells differentiated along the neural lineage in vitro are an attractive model system. Here we have developed ES cell lines that are suitable for inserting transgenes at a single chromosomal site. ES cell line CE1 (for Cassette Exchange) contains one "acceptor" module (CE1) that allows for efficient double lox targeting. The site is also permissive for gene expression in neural progenitor cells, as well as differentiated neurons and glia. Line CE2 was derived by swapping a puromycin resistance cassette into CE1. Neural progenitors derived from this line are puromycin-resistant. A beta-actin/GFP expression cassette was inserted into the CE1 site to create CE3. The CE3 cell line was differentiated into neural cells and displayed strong EGFP expression in neural progenitors, differentiated neurons and glia. Differentiated CE3 ES cells (4-/4+ RA) were transplanted into the injured rat somatosensory cortex where many of the transplanted cells survived and differentiated into neurons expressing GFP. This strategy for creating sets of transgenic lines with multiple cassettes inserted into a single chromosomal site provides a powerful tool for studying development and function of ES cell-derived neural cells. Many of these will be useful in transplantation research. PMID- 12591159 TI - Abnormal PI3 kinase/Akt signal pathway in vagal afferent neurons and vagus nerve of streptozotocin-diabetic rats. AB - The PI3 (phosphatidylinositol-3) kinase/Akt (protein kinase B) signal pathway is involved in the molecular signaling that regulates retrograde axonal transport of neurotrophins in the nervous system. Previous work showed that a reduced retrograde axonal transport of endogenous nerve growth factor (NGF) and neurotrophin-3 (NT-3) in the vagus nerve of diabetic rats occurred in the presence of normal production of neurotrophins and neurotrophin receptors. To assess the potential involvement of an impaired PI3 kinase/Akt signal pathway in the diabetes-induced reduction in retrograde axonal transport of neurotrophins in the vagus nerve, we characterized diabetes-induced changes in the PI3 kinase/Akt signal pathway in the vagus nerve and vagal afferent neurons. Control and streptozotocin (STZ)-induced diabetic rats with a duration of 16 weeks, kinase assays, Western blotting, and immunocytochemistry were used to show that diabetes resulted in alterations in activity and protein expression of the PI3 kinase/Akt signal pathway in the vagus nerve and vagal afferent neurons. Diabetes caused a significant decrease in enzymatic activity of PI3 kinase and Akt (52 and 36% of control, respectively) in the vagus nerve. The reduced enzymatic activity was not associated with decreased protein expression of the p85 subunit of PI3 kinase, Akt and phosphorylation of Akt (ser473). In contrast, there was a significant increase in the phosphorylation of p70s6 kinase (thr421/ser424) along with a normal protein expression of p70s6 kinase in the vagus nerve of diabetic rats. However, diabetes induced an overall decrease in immunoreactivity of the p85 subunit of PI3 kinase, phospho-Akt (ser473) and phospho-p70s6/p85s6 kinase (thr421/ser424) in vagal afferent neurons. Thus, impaired PI3 kinase/Akt signal pathway may partly account for the reduced retrograde axonal transport of neurotrophins in the vagus nerve of STZ-induced diabetic rats. PMID- 12591160 TI - Opposite reaction of ERK and JNK in ischemia vulnerable and resistant regions of hippocampus: involvement of mitochondria. AB - Delayed ischemic death of neurones is observed selectively in CA1 region of hippocampus at 3-4 days of reperfusion. Signals generated immediately during and after ischemia are further propagated by a variety of kinases, proteases and phosphatases. Tissue samples from dorsal (vulnerable) and abdominal (resistant) parts of gerbil hippocampi were collected to determine the activation state of key signaling molecules: Akt, Raf-1, JNK, ERK1/2 in the course of reperfusion after 5 min of global cerebral ischemia. Western blot analysis of phosphorylated forms of the kinases revealed persistent activation of JNK, being limited mostly to vulnerable CA1 region. On the contrary, activation of ERK, although observed transiently in both parts, was enhanced for a longer time in the abdominal hippocampus. The levels of the active/phosphorylated Akt and Raf-1 kinases did not change significantly during the recovery period. No significant correlation between postischemic JNK activation and c-Jun phosphorylation or its contribution to AP1-like complex formation was found. In contrast, the amount of active JNK linked with mitochondrial membranes was significantly increased and preceded neuronal death in CA1. In the same period of time the AP1 complex, augmented in CA1 region, did not appear to contain a classical c-Fos protein. These results are consistent with the theory that either long-lasting activation of JNK and/or contrasting ERK and JNK activities in critical time of reperfusion, contribute to selective apoptosis of CA1 neurons. This, in connection with the translocation of activated JNK to mitochondria and time/regional differences in AP1 binding protein complexes can affect final postischemic outcome. PMID- 12591161 TI - Expression of cytokines and cytokine receptors in the rat brain after kainic acid induced seizures. AB - We have previously shown that IL-6 protein levels are increased in cerebrospinal fluid in humans after recent tonic-clonic seizures with unchanged levels of IL 1beta and TNFalpha. Here we studied the expression of cytokines IL-6, LIF, IL 1beta and TNFalpha and cytokine receptors IL-6R, LIFR and Gp130 in the rat brain after kainic acid-induced status epilepticus using Northern blot analysis and in situ hybridization histochemistry. After seizures, IL-6 mRNA was induced in the hippocampus, cortex, amygdala and meninges, and IL-6R was up-regulated in the hippocampus. LIF was up-regulated in the hippocampus, cortex and meninges after seizures, and LIFR mRNA was induced in the hippocampus and cortex. Gp130 was constitutively expressed in the brain. After seizures, Gp130 transcription was rapidly induced in the meninges. In thalamus, cortex, amygdala and hippocampus Gp130 mRNA was induced in a delayed fashion. IL-1beta transcription was induced in the temporal lobe cortex and thalamus, and TNFalpha in the hippocampus. In general, the cytokine and their receptor mRNA levels were low in intact rat brain, but were induced by seizures. Since IL-6 and LIF transcripts were induced in the meninges after seizures, the protein products of these transcripts may be more readily released in cerebrospinal fluid after seizures. In addition, the activity of IL-6 and LIF signaling pathways may be influenced by increased expression of their receptors after seizures. PMID- 12591162 TI - The mu opioid agonist DAMGO stimulates cAMP production in SK-N-SH cells through a PLC-PKC-Ca++ pathway. AB - The mu-opioid agonist DAMGO exerts a dual activity on cAMP production in SK-N-SH neuroblastoma cells. While the classic inhibitory effect was prevented by pretreating the cells with pertussis toxin (PTX), the stimulatory activity was PTX-resistant. The stimulatory effect was abolished by the selective phospholipase C (PLC) blocker U-73122, by the selective protein kinase C (PKC) blocker chelerythrine and by the calcium-channels blockers Ni++, Co++ and Cd++. Hence, it is suggested that the opioid receptor activates PLC (probably through Gq GTP-binding proteins), to mobilize PKC, that positively modulates calcium channels in the plasma membrane; the entry of Ca++ into the cells stimulates calcium-activated adenylyl cyclases to produce cAMP. PMID- 12591163 TI - Whisker stimulation-dependent translation of FMRP in the barrel cortex requires activation of type I metabotropic glutamate receptors. AB - Fragile X syndrome is a common inherited cause of mental retardation that results from the absence of the Fragile X Mental Retardation Protein (FMRP), an RNA binding protein thought to regulate translation of bound mRNAs, including its own. Previous studies in our laboratory have shown that FMRP expression increases in the barrel cortex of the rat after unilateral whisker stimulation, a model of experience dependent plasticity. This increase in protein is restricted to sub cellular fractions enriched for synaptic or poly-ribosomal complexes. Here, we demonstrate that these increases are not accompanied by a change in FMR-1 mRNA levels and that they are blocked by the protein synthesis inhibitor cycloheximide in a dose dependent manner. Whisker stimulation dependent expression of FMRP is also abolished by pharmacological blockade of either NMDA receptors (MK-801, 0.25 mg/kg) or type I metabotropic glutamate receptors (AIDA, 5 mg/kg). In primary cortical neurons, activation of type I mGluRs leads to an increase in FMRP expression that is not effected by blockade of NMDA receptors. Taken together, these studies show that experience regulates FMRP production in vivo at the level of translation and supports a role for FMRP in metabotropic glutamate receptor mediated synaptic plasticity. PMID- 12591164 TI - Identification of a single QTL, Mptp1, for susceptibility to MPTP-induced substantia nigra pars compacta neuron loss in mice. AB - The loss of substantia nigra pars compacta (SNpc) neurons seen in idiopathic Parkinson's disease is hypothesized to result from a genetic susceptibility to an unknown environmental toxin. MPTP has been used as a prototypical toxin, since exposure to this drug results in variable SNpc cell death in several vertebrate species, including man and mouse. Previously, we have shown that C57BL/6J mice are sensitive to this compound, while Swiss-Webster mice are resistant. In this study, we intercrossed these mouse strains to map quantitative trait loci (QTL) for MPTP sensitivity. Using genome wide PCR analysis, we found that a single major QTLs, Mptp1, located near the distal end of chromosome 1 between D1Mit113 and D1Mit293, accounts for the majority of the strain sensitivity to MPTP. PMID- 12591165 TI - Molecular characterization of new polymorphisms at the beta2, alpha1, gamma2 GABA(A) receptor subunit genes associated to a rat nonpreferring ethanol phenotype. AB - Recent preclinical and clinical studies have indicated a possible involvement of the genes encoding for the GABA(A) receptor subunits alpha6, beta2, alpha1 and gamma2 in the genetic susceptibility to alcohol abuse. We have recently found an (R) to (Q) mutation in codon 100 of the alpha6 GABA(A) subunit, that segregated in a rat line selectively bred for its voluntary ethanol aversion, Sardinian alcohol nonpreferring (sNP), but not in their Sardinian alcohol preferring (sP) counterpart, selected for its ethanol preference. In the present study the molecular composition of other GABA(A) subunits (beta2, alpha1 and gamma2) were analyzed in order to further investigate the involvement of the GABA(A) receptors in the genetic predisposition to voluntary alcohol intake. Automated sequencing analysis indicated the presence of six new silent substitutions (289 T-->C in the beta2 gene; 115 G-->A in the alpha1 gene; 157 G-->A, 174 C-->T, 347 A-->G and 385 A-->T in the gamma2 gene), in sNP but not in sP rats. These polymorphisms were linked to the alpha6 R100Q mutation previously described in sNP rats. The strict association between the alpha6 point mutation and the new polymorphisms found in the beta2, alpha1 and gamma2 genes, demonstrate that such genes belong to the same cluster and are inherited together in the rat. These results sustain the synteny for these clusters between the rodent and human genomes, and suggest that mutated GABA(A) beta2, alpha6, alpha1 and gamma2 subunit genes might contribute to the expression of an ethanol nonpreferring phenotype in a rat line that voluntarily avoids alcoholic solutions. PMID- 12591166 TI - Sensory neuron proteins interact with the intracellular domains of sodium channel NaV1.8. AB - Voltage-gated sodium channels initiate and propagate action potentials in excitable cells. The tetrodotoxin-resistant Na(+) channel (Na(V)1.8/SNS) is expressed in damage-sensing neurons (nociceptors) and plays an important role in pain pathways. Expression of high levels of functional Na(V)1.8 in heterologous cells has proved problematic, even in the presence of known sodium channel accessory beta-subunits. This suggests that other regulatory proteins are required for normal levels of Na(V)1.8 expression. Here we report the use of a yeast two-hybrid system and a rat dorsal root ganglion cDNA library to identify 28 different clones encoding proteins which interact with intracellular domains of Na(V)1.8. Many clones are expressed at high levels in small diameter DRG neurons as judged by in situ hybridization. Interacting proteins include cytoplasmic elements and linker proteins (e.g. beta-actin and moesin), enzymes (e.g. inositol polyphosphate 5-phosphatase and TAO2 thousand and one protein kinase), channels and membrane-associated proteins (voltage-dependent anion channel VDAC3V and tetraspanin), as well as motor proteins (dynein intermediate and light chain) and transcripts encoding previously undescribed proteins. Immunoprecipitation (pull-down) assays confirm that some of the proteins interact with, and may hence regulate, Na(V)1.8 in vivo. PMID- 12591168 TI - Neuroendocrine-immune interactions. AB - Over the past 15 years, the mechanisms by which the brain communicates with the immune system and the immune system with the brain have been elucidated. Increasingly, we are beginning to realize that cytokines within the brain are no scientific oddity, but rather are an important component of normal brain functioning. They are able to alter behaviour, memory, stress responses, brain injury and are increasingly being linked to psychiatric disorders. Hormones, peptides and neurotransmitters regulate the immune system. Understanding the complex interactions of these systems is a challenge for the 21st century. PMID- 12591167 TI - Molecular cloning and characterisation of a novel GABAB-related G-protein coupled receptor. AB - Using a homology-based bioinformatics approach we have analysed human genomic sequence and identified the human and rodent orthologues of a novel putative seven transmembrane G protein coupled receptor, termed GABA(BL). The amino acid sequence homology of these cDNAs compared to GABA(B1) and GABA(B2) led us to postulate that GABA(BL) was a putative novel GABA(B) receptor subunit. The C terminal sequence of GABA(BL) contained a putative coiled-coil domain, di-leucine and several RXR(R) ER retention motifs, all of which have been shown to be critical in GABA(B) receptor subunit function. In addition, the distribution of GABA(BL) in the central nervous system was reminiscent of that of the other known GABA(B) subunits. However, we were unable to detect receptor function in response to any GABA(B) ligands when GABA(BL) was expressed in isolation or in the presence of either GABA(B1) or GABA(B2). Therefore, if GABA(BL) is indeed a GABA(B) receptor subunit, its partner is a potentially novel receptor subunit or chaperone protein which has yet to be identified. PMID- 12591169 TI - Gender matters: estrogen protects from cardiac hypertrophy. AB - A recent study shows that estrogens protect the female heart from the hypertrophy resulting from disturbance of myocardial calcium metabolism. Estrogens can inhibit cardiac hypertrophy by counteracting hypertension, by direct effects on the heart and by triggering the release of cardioprotective factors. However, because the hypertrophic response to increased cardiac load is primarily an adaptive process, the inhibition of hypertrophy might not always be beneficial. Estrogen therapy could interfere with the utilization of the larger hypertrophic reserve in the female heart, and predispose the female heart to systolic dysfunction. PMID- 12591170 TI - Progesterone in gestational diabetes mellitus: guilty or not guilty? AB - Insulin resistance is one of the metabolic changes in pregnancy, but only a fraction of women develop overt impaired glucose tolerance or frank diabetes. Most women are able to compensate this altered metabolic state by increasing the amount of insulin produced by the pancreatic beta cells. Progesterone might well be the key to the development of gestational diabetes. Previously high progesterone levels have already been shown to be correlated with the development of glucose abnormalities in pregnancy and now, in a new paper, progesterone receptor-knockout mice are found to have improved glucose tolerance. These mice showed increased insulin secretion, which is probably linked to the presence of increased numbers of beta cells in their pancreas. Is progesterone therefore the 'ultimate bad guy', prohibiting normal adaptation of the pancreatic beta-cell reserve during pregnancy? PMID- 12591171 TI - The role of chromogranin A and the control of secretory granule genesis and maturation. PMID- 12591176 TI - Alternative splicing: a novel mechanism to fine-tune the expression and function of the human AT1 receptor. AB - Activation of the angiotensin II type 1 (AT(1)) receptor is closely involved in the pathogenesis of cardiovascular diseases; therefore, aberrant regulation of the production of this receptor might play a role in these disorders. Currently, there is strong evidence to suggest that the predominant mechanism regulating the number of AT(1) receptors is the modulation of mRNA stability. Here, we discuss the importance of alternative splicing as an additional post-transcriptional mechanism regulating human AT(1) receptor number and function. PMID- 12591175 TI - Anorectic brainstem peptides: more pieces to the puzzle. AB - Eating a meal is a mechanical process involving autonomous pathways that relay sensory and motor information between the whole length of the digestive tract and the central nervous system. This circuitry is able to initiate and terminate the meal, primarily by gut-brainstem-gut reflex arcs, and is independent of the caloric content of a meal. However, as part of our ability to regulate body weight over time, we must be able to modulate the amount of energy that we take in as food and the amount of energy that we expend. Thus, the gut-brainstem axis must be coupled to other systems that take account of factors such as food availability and preference, changing energy requirements and our social habits. Here, we review the importance of the brainstem nucleus of the tractus solitarius as a site of integration and the routes by which it connects the gut-brainstem axis with regulatory neuronal and endocrine networks that allow for strict body weight management. PMID- 12591177 TI - AT4 receptor is insulin-regulated membrane aminopeptidase: potential mechanisms of memory enhancement. AB - Although angiotensin IV (Ang IV) was thought initially to be an inactive product of Ang II degradation, it was subsequently shown that the hexapeptide markedly enhances learning and memory in normal rodents and reverses the memory deficits seen in animal models of amnesia. These central nervous system effects of Ang IV are mediated by binding to a specific site, known as the AT(4) receptor, which is found in appreciable levels throughout the brain and is concentrated particularly in regions involved in cognition. This field of research was redefined by the identification of the AT(4) receptor as the transmembrane enzyme, insulin regulated membrane aminopeptidase (IRAP). Here, we explore the potential mechanisms by which Ang IV binding to IRAP leads to the facilitation of learning and memory. PMID- 12591178 TI - Transcription factors direct the development and function of pancreatic beta cells. AB - Transcription factors orchestrate intricate pathways of cellular growth and differentiation by regulating the rate of transcription of an array of genes. Genetic and biochemical studies have begun to unravel the complex cascade of factors that controls the proliferation and differentiation of cells in the developing pancreas. The specific pathway leading to the development of the insulin-secreting beta cell has been a focus of many of these studies because an understanding of the transcription factors governing this pathway will be crucial to the engineering of new beta cells to cure diabetes. In recent years, the number of transcription factors that has been implicated in beta-cell differentiation and function has grown considerably. Here, we outline the known role of transcription factors in beta-cell development, and describe how these factors form a network of gene activation signals that mediates insulin transcription. PMID- 12591179 TI - Thyroid hormone receptors: lessons from knockout and knock-in mutant mice. AB - The genes encoding thyroid hormone receptor alpha and beta (TRalpha and TRbeta) encode four thyroid hormone receptors and four variant isoforms with antagonistic properties. Because of this complexity, numerous models of TR mutation have been developed to understand the functions of specific receptors. In total, 13 mutant strains are now available. Phenotype analysis has shown that the two genes serve distinct functions: TRalpha is crucial for postnatal development and cardiac function, whereas TRbeta mainly controls inner ear and retina development, liver metabolism and thyroid hormone levels. These mouse mutant strains also provide us with the unique opportunity to address the respective contribution of each receptor isoform and isotype in vivo and highlight the in vivo importance of the ligand-independent function of the TR gene products. PMID- 12591182 TI - The management of alcoholic liver disease. PMID- 12591180 TI - Transduction pathway of anti-Mullerian hormone, a sex-specific member of the TGF beta family. AB - Anti-Mullerian hormone (AMH), also called Mullerian inhibiting substance, is a member of the transforming growth factor beta (TGF-beta) family that represses the development and function of reproductive organs. Not for nothing did Professor Alfred Jost, who first discovered its existence, christen it 'hormone inhibitrice'! Anti-Mullerian hormone is thought to exert its effects through two membrane-bound serine/threonine kinase receptors, type 2 and type 1. Upon ligand binding, these drive receptor-specific cytoplasmic substrates, the Smad molecules, into the nucleus where they act as transcription factors. A type 2 receptor specific for AMH was cloned through its homology with receptors of TGF beta family members; the identity of the type 1 receptor(s) is controversial. Three type 1 receptors for bone morphogenetic proteins (BMPs) are possible candidates, each, not surprisingly, activating BMP-specific Smad molecules, Smads 1, 5 and 8. Each receptor could be involved, depending on the cellular context. To date, AMH signaling has been explored through BMP-specific genes, because a reporter gene related to a physiological AMH function and upregulated by the hormone has not yet been tested in a cell line strongly expressing the AMH receptor(s). PMID- 12591183 TI - Hereditary hemochromatosis: update for 2003. PMID- 12591184 TI - Current management of primary biliary cirrhosis and primary sclerosing cholangitis. PMID- 12591185 TI - Liver fibrosis -- from bench to bedside. PMID- 12591186 TI - Current management of portal hypertension. PMID- 12591187 TI - Ascites and hepatorenal syndrome in cirrhosis: pathophysiological basis of therapy and current management. PMID- 12591188 TI - Management of chronic hepatitis B. PMID- 12591189 TI - Management of hepatitis C. PMID- 12591190 TI - Living donor liver transplantation. PMID- 12591191 TI - Hepatocellular carcinoma: present status and future prospects. PMID- 12591192 TI - Two-step (culture and PCR) diagnostic approach for differentiation of non-T. foetus trichomonads from genitalia of virgin beef bulls in Argentina. AB - Preputial fluids from 567 virgin Angus and Hereford bulls, 1-2 years old, were inoculated into Sutherland medium, and approximately 8.4% produced cultures with a protozoan suggestive of Tritrichomonas foetus. Under brightfield microscopy, large numbers of single-celled motile organisms with multiple anterior flagellae, a posterior flagellum, axostyle, and a visible undulating membrane were detectable. Motility was jerky and rolling, as described for T. foetus. Air-dried smears of cultures stained with Giemsa or Diff-Quick + iodine revealed an organism similar to T. foetus, although somewhat more rounded. Several organisms appeared to have four anterior flagellae. Scanning electron microscopy (5000x) of representative samples revealed four anterior flagellae on most organisms, and an axostyle that was consistently longer than that seen in T. foetus. Using pan trichomonal primers and T. foetus-specific primers in a polymerase chain reaction (PCR) assay, amplification products of 372bp were detected in all virgin bull isolates, but only with the pan-trichomonal primers. Positive control isolates of T. foetus yielded amplification products of the expected size (372 and 347bp) with the two sets of primers, respectively. We conclude that these protozoa are not T. foetus, and note the similarity of these findings with those reported earlier in North American beef cattle. Because in several countries there is no legal treatment for bovine trichomonosis, veterinarians recommend slaughter of bulls with positive preputial cultures. The existence of easily mis-identified non-T. foetus trichomonads in the bovine prepuce suggests that the current "gold standard" diagnostic test (culture of preputial scrapings or washings) should be augmented with a more specific confirming test, such as the PCR employed in this study. PMID- 12591193 TI - Evaluation of PCR to detect Theileria parva in field-collected tick and bovine samples in Tanzania. AB - The ability of PCR to detect infections of Theileria parva, the cause of East Coast Fever, in field-collected tick and bovine samples from Tanzania was evaluated. PCR-detected infection prevalence was high (15/20, 75%) in unfed adult Rhipicephalus appendiculatus ticks that fed as nymphs on an acutely-infected calf, but low (22/836, 2.6%) in unfed adult R. appendiculatus collected from field sites in Tanzania. Tick infection prevalence was comparable to that in previous studies that used salivary gland staining to detect T. parva infection in field-collected host-seeking ticks. Of 282 naturally-exposed zebu calves, seven had PCR-positive buffy coat samples prior to detection of Theileria spp. parasites in stained buffy coat cells or lymph node biopsies. Evidence of Theileria spp. infections was detected in stained smears of lymph node biopsies from 109 calves (38.6%) and buffy coat samples from 81 (28.7%), while buffy coat samples from 66 (23.4%) were PCR-positive for T. parva. Implications of these findings for the sensitivity and specificity of the PCR are discussed. PMID- 12591194 TI - In vitro culture of Glugea sp. AB - Recently the low host specificity of some microsporidians has been demonstrated and it has been indicated that many of these micro-organisms could be transmitted from invertebrates to mammals and adapt to changes in temperature. In this work, we demonstrate the first successful in vitro culture of a fish microsporidia of the genus Glugea on larval cells of the mosquito Aedes albopictus at 28 degrees C, and we show ultrastructural aspects of the different life cycle stages. It was impossible on salmon cells CHSE-214 at 21 degrees C. This study will be valuable for further work in biochemistry and immunology in addition to chemotherapy for microsporidiosis humans and animals. PMID- 12591195 TI - Molecular differentiation of Hypoderma bovis and Hypoderma lineatum (Diptera, Oestridae) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). AB - The most variable region of the cytochrome oxidase I (COI) gene of Hypoderma bovis(1) and Hypoderma lineatum(2) (Diptera, Oestridae) was amplified by PCR and the amplicons were sequenced and analysed. PCR products were digested with three restriction enzymes, namely BfaI, HinfI and TaqI, providing informative profiles. H. bovis and H. lineatum sequences revealed an inter-specific variation rate of 8.5%, and an intra-specific variation rate of 0.87 and 0.29%, respectively. The results showed that the COI gene region examined was useful for the differentiation of H. bovis and H. lineatum and that a PCR-RFLP assay is a practical tool for their identification, offering additional diagnostic and epidemiological instruments for the study of cattle grub infestation. PMID- 12591196 TI - Efficacy of eprinomectin pour-on against gastrointestinal nematode infections in sheep. AB - Two field trials were conducted in two farms (farms A and B) in southern Italy, to assess the efficacy of eprinomectin applied topically at the dose rate of 500 micro g/kg to sheep with naturally occurring infections of gastrointestinal nematodes (GIN). The nematode population determined by necropsy consisted of Teladorsagia circumcincta, Haemonchus contortus, Trichostrongylus vitrinus, T. capricola, Nematodirus sp., and Chabertia ovina in sheep from farm A, and of T. circumcincta, T. vitrinus, T. capricola, T. colubriformis, and C. ovina in sheep from farm B. In each farm, 42 female sheep were assigned to a eprinomectin treated group (E-group) and a control untreated group (C-group) of 21 animals each. On farm A, the percentage reductions in strongyle faecal egg counts from E group compared to C-group were 99.1% on day 10; 97.4% on day 30; and 67.0% on day 60. On farm B, on the same days, they were 95.4, 84.9, and 69.4%, respectively. In the course of the two trials, eprinomectin was well tolerated by all the animals with no adverse reactions following the topical treatment. PMID- 12591197 TI - Enhancing immunity to nematode parasites in single-bearing Merino ewes through nutrition and genetic selection. AB - The effectiveness of protein supplementation and genetic selection to enhance the resistance of periparturient Merino ewes to infection from gastrointestinal parasites was tested in a replicated grazing experiment. One hundred and twenty ewes from lines selected for increased resistance (R) to Haemonchus contortus or at random (C) were subjected to one of the three supplement groups that provided 0 or 250 g per day cottonseed meal for 5 weeks prior to, or for 6 weeks after the start of lambing. Faecal egg counts (FEC) of R ewes were consistently lower than those of C ewes but both groups exhibited a periparturient rise in FEC. Supplementation during the pre-partum period reduced FEC and increased ewe body weight gain. The benefits of pre-partum supplementation in reducing FEC continued to be apparent up to 10 weeks after supplementation ceased. There was a strong suggestion that the benefits to parasite resistance from protein supplementation were greatest in C ewes. Wool growth rates (15%) and birth weights (5%) were greater for C ewes but differences between the lines for lamb body weight had disappeared by day 97. The greatest benefit to resistance from protein supplementation was observed when ewes were experiencing a loss of maternal body weight. Conversely, no benefits to resistance were observed when ewes had moderate (78-107 g per day) rates of maternal weight gain. These results suggest that increased resistance as a result of protein supplementation is dependent on the prevailing supply and demand for scarce nutrients such as metabolisable protein (MP). Both genetic selection and protein nutrition are effective strategies to enhance host resistance to nematode infection during the periparturient period. PMID- 12591198 TI - The development of Trichostrongylus colubriformis larvae on a range of herbage species or on plots of differing topographical aspect. AB - Five "contaminations", where faeces containing Trichostrongylus colubriformis eggs were deposited on pasture and serially recovered, were used to compare the rate of decline of faecal mass and larval development. In the first three contaminations, faeces from a common source were deposited on swards of browntop (Agrostis capillaris cv Grasslands Muster), ryegrass (Lolium perenne cv Grassland Nui), white clover (Trifolium pratense cv Grassland Tahora), or onto bare ground in the late spring, summer or autumn. The last two contaminations were done on the north facing aspect or south facing aspect of hill country pastures in summer and autumn. Number of free-living nematodes (first- and second-stage larvae (L(1) and L(2)) and soil dwelling nematodes) and third stage larvae (L(3)) recovered from faeces were counted. In spring there was a significant (P<0.01) effect of sward type on the mass of faeces remaining, with greatest mass remaining on browntop and ryegrass 28 days later, and less on bare ground and white clover. In summer there were more (P<0.05) faeces remaining on browntop than on other herbages which had little faeces remaining and which did not differ one from another. In autumn there was a rapid decline in faecal mass. All faeces were gone from white clover and ryegrass swards by day 10 and from browntop and bare ground by day 14. The number of free-living nematodes did not differ markedly between seasons, ranging from 5 to 8.5% of eggs deposited. The number of L(3) recovered was low in spring ( approximately 0.4% of eggs deposited) and did not differ between swards. In summer, more (P<0.05) L(3) were recovered from faeces deposited on swards of ryegrass and white clover than from bare ground or browntop. Most L(3) were recovered from days 7 to 14 ( approximately 1.3% of eggs deposited). In the autumn, low numbers of L(3) were recovered from browntop on day 3 and ryegrass on day 7 (0.2% of eggs deposited) with virtually no L(3) recovered from faeces placed on white clover or bare ground. There were significant (P<0.001) effects of aspect on the amount of faecal mass remaining in both summer and autumn with less faeces remaining on the south facing aspect than on the north. This was particularly evident during the summer when virtually all of the faeces were intact on the north facing aspect but only 40% was remaining on the south on day 28. In the autumn, while faeces were completely gone from both aspects by day 28 but there were less (P<0.05) faeces remaining on the south facing aspect from days 3 to 18 than from the north. There was no aspect effect in either season on the number of free-living nematodes recovered which averaged 8-11% of eggs deposited. In both seasons a greater number of L(3) were recovered from faeces on the south facing aspect than on the north, particularly 3-10 days after faecal deposition. In summer the rise in L(3) recovered in faeces was more rapid on south facing aspect than on the north but both attained a maximum level of approximately 4% of eggs deposited. In autumn on day 3 there was a rapid rise on south facing aspect to approximately 21% of eggs deposited followed by a gradual decline on day 10 while on the north facing aspect numbers of L(3) recovered only attained 10% of eggs deposited. PMID- 12591199 TI - Efficacy of selamectin administered topically in the treatment of feline otoacariosis. AB - The efficacy of a novel avermectin, selamectin (Stronghold, Pfizer), was evaluated against naturally acquired aural infestations of Otodectes cynotis. Selamectin was administered topically in a single spot to the skin of each animal's back at the base of the neck in front of the scapulae at a minimum dosage of 6mgkg(-1). Thirty cats of a cattery of 120 animals with an endemic infestation were treated on days 0 and 30. Including untreated control cats was therefore not possible because of animal welfare consideration. Clinical examination and visualization of mites by otoscopic examination of the external ear canal and microscopic examination of aural debris/exudate were performed twice weekly from days 0 to 30. On day 0 (and day 28 if the ears canals were erythematous), a swab was taken from each canal and sent to bacteriology and mycology units for detection and identification of bacteria (Staphylococcus mainly) and yeast (Malassezia pachydermatis only). Selamectin was safe, parasites were killed before day 3 and eliminated before day 17. Selamectin was 100% effective against natural aural infestations, even if erythema (26-33% of cats) and scratch reflex (23-40% of cats) persisted for 2 weeks after the cats tested negative for mites. PMID- 12591200 TI - Comparative speed of kill between nitenpyram, fipronil, imidacloprid, selamectin and cythioate against adult Ctenocephalides felis (Bouche) on cats and dogs. AB - Speed of kill and percentage kill of nitenpyram (CAPSTAR) was compared to fipronil (Frontline) spot-on), imidacloprid (Bayvantage/Advantage), selamectin (Stronghold/Revolution) and cythioate (Cyflee) against adult fleas on cats and dogs 3 and 8h post-treatment. Selamectin was used on dogs only; cythioate was used on cats only. Groups of eight cats and eight dogs (four males and four females each) were experimentally infested with 100 unfed fleas 1 day prior to treatment with the test products. One group of cats and one group of dogs served as control. Fleas were collected from four cats and four dogs in each group (two males and two females) by combing 3h after treatment, the remaining four cats or dogs were combed 8h after treatment. In cats cythioate treatment resulted in a mean efficacy of 62.4 and 97.4% at 3 and 8h post-treatment, respectively. Imidacloprid efficacy at the same times was 26.9 and 82.8%, whereas fipronil efficacy was 24.3 and 62.6% efficacy, respectively. In dogs mean efficacy 3 and 8h after treatment with selamectin was 39.7 and 74.4%; with imidacloprid efficacy was 22.2 and 95.7%, respectively and 35.9 and 46.5%, respectively after treatment with fipronil. Nitenpyram was 100% effective in cats and 99.1% effective in dogs within 3h of treatment and 100% effective in cats and dogs within 8h. PMID- 12591201 TI - Comment on "A comparison of persistent anthelmintic efficacy of topical formulations of doramectin, eprinomectin, ivermectin and moxidectin against naturally acquired nematode infections of beef calves" and problems associated with mechanical transfer (licking) of endectocides in cattle. PMID- 12591202 TI - Antigenic characterization of foot-and-mouth disease virus serotype Asia1 field isolates using polyclonal and monoclonal antibodies. AB - Foot-and-mouth disease virus (FMDV) serotype Asia1 field isolates (n = 100) were compared using a panel of 11 monoclonal antibodies (Mab) in sandwich ELISA. The majority (over 89%) of the isolates showed either homologous (76% and above reactivity) or reduced affinity (20-75% reactivity) for the Mabs 2A, 13, 40, 34 and 81, suggesting that these Mab binding epitopes are conserved, whereas a more variable reactivity was observed for the Mabs B3, 1A, 24, 72, 82 and 89. Polyclonal relationship ('r' value) of the field isolates in liquid phase blocking (LPB) ELISA was examined, and the mean 'r' value was 0.62 relative to vaccine virus IND 63/72. Some of the field isolates (n = 34) were tested in virus neutralization test (VNT) and showed an 'r' value of >0.40. Although a minor antigenic difference was observed in the Mab profiling study, there has not been large antigenic divergence between reference virus and field viruses, thereby providing evidence of wide antigenic coverage of the vaccine strain. PMID- 12591203 TI - Strain specificity of the immune response of pigs following vaccination with various strains of porcine reproductive and respiratory syndrome virus. AB - The primary objective of the study was to determine strain specificity of the immune response of pigs following vaccination with selected strains of porcine reproductive and respiratory syndrome virus (PRRSV). The experimental design included five groups (I through V, six pigs per group) free of antibody for PRRSV at the beginning of the experiment (day 0). On day 0, groups III, IV, and V were vaccinated with attenuated versions of PRRSV strains 8, 9, and 14, respectively. On day 21, the immunity of group II (non-vaccinated/challenged controls) and groups III, IV, and V was challenged by exposing each pig to a composite of the virulent versions of these same three strains. On day 35, all pigs, including non vaccinated/non-challenged pigs of group I, were necropsied. Lungs and selected lymph nodes were examined for lesions. Serum samples obtained at weekly intervals throughout the study and lung lavage fluids obtained at necropsy were tested for the presence of PRRSV and its strain identity. Before challenge the strain of PRRSV identified in the sera of vaccinated pigs was always that with which the particular pig had been vaccinated (i.e. homologous strain), whereas, with one exception, only heterologous strains were identified after challenge. This apparent strain exclusion as a result of vaccination was statistically significant (P = 0.004). The tendency for heterologous strains to predominate after challenge suggests that a pig's immune response to PRRSV has some degree of strain specificity. Whether this finding has any clinical relevance in regard to immunoprophylaxis remains to be determined. PMID- 12591204 TI - Comparative safety and efficacy of attenuated single-strain and multi-strain vaccines for porcine reproductive and respiratory syndrome. AB - The objective of this study was to compare the efficacy and safety of single strain and multi-strain vaccines for the prevention of the respiratory facet of porcine reproductive and respiratory syndrome. The study comprised six groups of pigs (A through F, eight pigs per group). At the beginning of the study (Day 0) Groups C and D were vaccinated with a single-strain vaccine, and Groups E and F were vaccinated with a multi-strain vaccine. The multi-strain vaccine contained five attenuated strains of PRRSV including the strain used as the single-strain vaccine. On Day 28 Groups B (nonvaccinated/challenged control), D, and F were challenged with a highly virulent field strain of PRRSV that was unrelated to any of the strains used for vaccination. Group A was kept as a nonvaccinated/nonchallenged control. On Day 42 all pigs were necropsied. Their lungs and lymph nodes were examined for virus-induced changes. Serum samples obtained at weekly intervals during the study and lung lavage fluids obtained at necropsy were tested for the presence and titer of PRRSV. Serum samples were also tested for antibody. The presence and severity of clinical signs and lesions were the primary means by which vaccine efficacy and safety were evaluated. Both vaccines provided a high level of protective immunity to challenge. However, appreciable lymph node enlargement in pigs vaccinated with multi-strain vaccine, with or without subsequent challenge, raised a question as to its safety. Collectively these results indicate that both single-strain and multi-strain attenuated PRRSV vaccines can be effective immunogens, but additional studies in regard to safety are needed before multi-strain vaccines can be recommended for routine field use. PMID- 12591205 TI - Association between the porcine Escherichia coli F18 receptor genotype and phenotype and susceptibility to colonisation and postweaning diarrhoea caused by E. coli O138:F18. AB - Porcine postweaning Escherichia coli enteritis is a cause of significant morbidity and mortality in pigs worldwide, and effective prevention remains an unsolved problem. This study examined the correlation between susceptibility of pigs to experimental infection with an E. coli F18 strain and the porcine intestinal F18 receptor genotypes. Thirty-one pigs classified as either belonging to the susceptible or the resistant genotype were inoculated with cultures of an E. coli O138:F18 isolated from a pig with postweaning diarrhoea. Susceptibility to colonisation and diarrhoea was assessed by clinical observations, faecal shedding of the challenge strain, histopathology and microscopic adhesion tests. Ten of 14 (71.4%) genetically susceptible pigs and one of 17 (5.9%) resistant pigs developed diarrhoea attributable to the challenge strain. There was no difference in susceptibility between homozygotic and heterozygotic susceptible pigs. Faecal shedding of the challenge strain correlated with the genetic receptor profile. Twenty pigs examined immunohistochemically revealed focal to extensive small intestinal mucosal colonisation by E. coli O138:F18 in nine of 10 susceptible and three of 10 resistant pigs. Results of in vitro adhesion assays performed with F18 cells on enterocyte preparations from 24 pigs, showed complete concordance with the F18 genotypes. In conclusion, this study showed a high correlation between the porcine intestinal F18 receptor genotypes and susceptibility to disease. However, pigs of the resistant F18 receptor genotype were not entirely protected against intestinal colonisation by E. coli F18. PMID- 12591206 TI - Comparison of polymerase chain reaction and bacteriological culture for the diagnosis of sheep brucellosis using aborted fetus samples. AB - PCR assay has been shown to be a promising option for the diagnosis of brucellosis. However, few studies have been performed with field samples in order to evaluate the assay as a diagnostic tool. In this study, routine use of a species-specific PCR assay previously developed for the identification of Brucella cultures was assessed for the detection of Brucella DNA directly from the stomach contents of aborted sheep fetuses. The assay is based on the insertion sequence IS711 in the Brucella chromosome. In the study, during 3 successive lambing seasons (1998-1999, 1999-2000 and 2000-2001) 126 aborted fetus samples each from different flocks and locations were examined. Brucella strains were isolated from 39 (31%) of the samples and all of the strains were identified as Brucella melitensis by biochemical characteristics, agglutination with monospecific A and M sera and PCR. Thirty-seven of 39 B. melitensis isolates were biotyped as biotype 3, and 2 isolates as biotype 1. From 38 of 39 culture positive fetal stomach contents B. melitensis-specific DNA was detected by PCR. PCR was found negative in all of the culture negative samples. Compared with culture, sensitivity and specificity of PCR were determined as 97.4 and 100%, respectively. The results indicate that this PCR procedure has a potential for use in routine diagnosis of sheep brucellosis. PMID- 12591207 TI - Antigen recognition by rainbow trout (Oncorhynchus mykiss) of whole cell proteins expressed by Lactococcus garvieae when obtained directly from fish and under iron limited culture conditions. AB - Infection by Lactococcus garvieae has become a widely recognised problem associated with intensively cultured fish. Long-term control of fish infections may be possible by vaccination providing a suitable and efficacious epitope is expressed during production of cells used for vaccine preparation. The identification of novel vaccine candidates must, therefore, consider how the host species recognises and responds to bacterial cell components. L. garvieae was cultured in iron deficient, limited and haem iron enriched media and the whole cell proteins expressed under these conditions were compared with those expressed in bacteria extracted with Percoll gradients directly from spleen tissue of infected rainbow trout (Oncorhynchus mykiss). SDS-PAGE of the cell proteins showed the existence of several different electropherotypes according to the iron status of the culture media. Only minor differences in cell protein profile were detected in bacteria obtained directly from fish spleens, but when the electropherograms were analysed by Western blots using L. garvieae hyperimmune fish sera, several proteins could be identified that were expressed only when L. garvieae was growing in vivo. Siderophore could be detected in culture supernatant of iron deficient, limited and haem iron enriched media but not in media with higher nutrient concentrations. The siderophore could not be identified as a type of catechol or hydroxymate. Rainbow trout recognise proteins in the range of approximately 50-80 kDa for bacterial cells obtained without subculture from infected fish and culture conditions can influence protein profiles for this pathogen. PMID- 12591208 TI - Identification of glycosphingolipid binding sites for SEF21-fimbriated Salmonella enterica serovar Enteritidis in chicken oviductal mucosa. AB - In order to clarify the presence of glycosphingolipids (GSLs) receptors for Salmonella enterica serovar Enteritidis with SEF21 fimbriae, we analyzed neutral GSLs and gangliosides from chicken oviductal mucosa and investigated the binding of bacteria to neutral GSLs and gangliosides. Five types of neutral GSLs, designated as N-1 to N-5, and two types of gangliosides, designated as G-1 and G 2, were identified on the thin-layer chromatography (TLC) plates. In the bacterial binding assay on TLC, the fimbriated bacteria bound only to glucosylceramide (GlcCer) standard, N-1 having the same TLC mobility as GlcCer, GM3 standard and G-1 corresponding to GM3 in TLC mobility, but not to N-2, N-3, N 4, N-5, or G-2. These results suggest the presence of GlcCer (N-1) and ganglioside GM3 (G-1) on the epithelial surface of chicken oviductal tract which act as sites for adherence of SEF21-fimbriated S. Enteritidis. PMID- 12591209 TI - Purification and renaturation of membrane neuraminidase from Haemophilus parasuis. AB - Haemophilus parasuis, which causes polyserositis, polysynovitis, meningitis, septicemia, and pneumonia in pigs, has emerged as an increasing problem in modern swine production systems. Co-factors for and the pathogenesis of H. parasuis disease are not defined. One of the potential virulence factors of H. parasuis is its neuraminidase (sialidase). While purifying the H. parasuis neuraminidase from the membrane fraction, we developed a protocol to renature enzymatic activity after enzyme preparations were resolved electrophorectically in denaturing polyacrylamide gels. The H. parasuis neuraminidase co-resolved with recombinant neuraminidase of Vibrio cholera; thus its apparent molecular mass is 82 kilodalton (kDa). The H. parasuis neuraminidase was associated with the membrane fraction and the purification protocol removed over 99% of the H. parasuis cell protein while retaining over 90% of the neuraminidase activity. Purified protein will provide another avenue to clone the neuraminidase gene that has been refractory to cloning and the protocol will be a means to purify recombinant protein. PMID- 12591210 TI - Auditory amplification: outer hair cells pres the issue. AB - Active feedback mechanisms in the ears of mammals amplify and tune the auditory response to sound stimuli, allowing these organisms to enjoy both exquisite sensitivity and sharp frequency selectivity. A requisite component of this feedback pathway has recently been identified. Targeted deletion of pres, the gene that encodes the novel membrane-bound motor protein prestin, has revealed a hundred-fold loss in hearing sensitivity suggesting that it might be part of the elusive cochlear amplifier. PMID- 12591211 TI - Protein phosphatase 1 and memory: practice makes PP1 imperfect? AB - Long-lasting memories are most efficiently formed by multiple training sessions separated by appropriately timed intervals. A recent study revealed that expression of a transgene encoding an inhibitor of protein phosphatase 1 (PP1) in the forebrain enhanced memory formed during sub-optimal training. Thus, PP1 apparently constrains memory formation in the mouse. Furthermore, the report proposes that PP1 promotes forgetting. PMID- 12591212 TI - Receptor tyrosine kinase transactivation: fine-tuning synaptic transmission. AB - G-protein-coupled receptors generate signals that promote gene transcription through the 'transactivation' of receptor tyrosine kinases (RTKs) and activation of the mitogen-activated protein kinase (MAPK) cascade -- a process that involves RTK autophosphorylation and endocytosis. Pioneering work now suggests that D4 dopamine-receptor-mediated transactivation of the platelet-derived growth factor beta receptor has immediate effects on synaptic neurotransmission via Ca(2+) dependent inactivation of NMDA receptors. The demonstration of a physiological role for RTK transactivation in the CNS provides novel opportunities for understanding how aberrant dopamine signalling might contribute to cognitive and attention deficits associated with schizophrenia and attention-deficit hyperactivity disorder. PMID- 12591213 TI - When neurons form memories. AB - Although long-term memory is central among our cognitive functions, the search for a direct neurophysiological correlate to it has proven difficult. The formation of new memories depends on the hippocampus and adjacent cortex, but the final storage is thought to be in a widely distributed neocortical network. Recent experiments, using simultaneous recordings from hundreds of sites in monkey neocortex, have revealed the activation of such a distributed network -- probably reflecting the consolidation of long-term memory storage. PMID- 12591214 TI - Stem and progenitor cells: the premature desertion of rigorous definitions. AB - A current disturbing trend in stem cell biology is the abandonment of rigorous definitions of stem and progenitor cells in favor of more ambiguous, all encompassing concepts. However, recent studies suggest that there are consistent, functional differences in the biology of these two cell types. Admittedly, it can be difficult to harmonize the in vivo and in vitro functional differences between stem and progenitor cells. Nonetheless, these distinctions between cell types should be emphasized rather than ignored, as they can be used to test specific hypotheses in neural stem cell biology. PMID- 12591215 TI - In search of a better mouse test. AB - To elucidate pathways from specific genes to complex behaviors, assays of mouse behavior need to be valid, reliable and replicable across laboratories. Behavioral assays are proving to be as complex as the intricate cellular and molecular pathways that are the main interest of many mouse users. There is no perfect behavioral test, but we propose some aphorisms to stimulate discussion that is necessary for continued progress in task development. For maximal utility, a behavioral test should yield valid data for most of the commonly used inbred mouse strains. Tests of simple, ubiquitous behaviors usually yield meaningful data for most mice, especially when based on automated scoring or on simple physical measures that are likely to be replicable across laboratories. Extreme test scores resulting from non-performance on a task can inflate the apparent reliability of a test, and devious adaptations to a task can undermine its validity. The optimal apparatus configuration for certain genetic or pharmacological analyses might depend on the particular laboratory environment. Despite our best efforts, the mice will continue to win some innings. PMID- 12591216 TI - Folate and homocysteine metabolism in neural plasticity and neurodegenerative disorders. AB - Folate is a cofactor in one-carbon metabolism, during which it promotes the remethylation of homocysteine -- a cytotoxic sulfur-containing amino acid that can induce DNA strand breakage, oxidative stress and apoptosis. Dietary folate is required for normal development of the nervous system, playing important roles regulating neurogenesis and programmed cell death. Recent epidemiological and experimental studies have linked folate deficiency and resultant increased homocysteine levels with several neurodegenerative conditions, including stroke, Alzheimer's disease and Parkinson's disease. Moreover, genetic and clinical data suggest roles for folate and homocysteine in the pathogenesis of psychiatric disorders. A better understanding of the roles of folate and homocysteine in neuronal homeostasis throughout life is revealing novel approaches for preventing and treating neurological disorders. PMID- 12591217 TI - Role of dendritic synapse location in the control of action potential output. AB - How effective are synaptic inputs made at different dendritic sites in the control of action potential output? Recent technical advances have allowed direct observation of the amplitude and time course of excitatory synaptic inputs generated at defined dendritic sites. These findings indicate that contrasting mechanisms exist to enhance the impact of dendritic excitatory inputs to pyramidal neurons of the neocortex and hippocampus. Furthermore, multiple sites of synaptic integration have been revealed, suggesting that proximal and distal inputs are likely to be integrated in different ways. Taken together, these data emphasize the cell- and input-specific strategies used by neurons to enhance the impact of dendritic inputs on action potential generation. PMID- 12591218 TI - Spine motility: a means towards an end? AB - From the first glimpse of moving spines half a decade ago, the prevailing view has been that spine contortions and wiggling, especially during development, maximize encounters with presynaptic growth cones or synaptic boutons. Other new evidence has revealed that spines continue to be motile even after they settle on a presynaptic partner and form a synapse. We present the evidence for each view, and discuss how spines with synapses could move relative to their apparently stable presynaptic partners. Thus, spine motility might not simply be a means towards an end of synapse formation, but could continue, albeit at a lower rate, during synapse turnover after development ends. PMID- 12591219 TI - Bursts as a unit of neural information: selective communication via resonance. AB - What is the functional significance of generating a burst of spikes, as opposed to a single spike? A dominant point of view is that bursts are needed to increase the reliability of communication between neurons. Here, we discuss the alternative, but complementary, hypothesis: bursts with specific resonant interspike frequencies are more likely to cause a postsynaptic cell to fire than are bursts with higher or lower frequencies. Such a frequency preference might occur at the level of individual synapses because of the interplay between short term synaptic depression and facilitation, or at the postsynaptic cell level because of subthreshold membrane potential oscillations and resonance. As a result, the same burst could resonate for some synapses or cells and not resonate for others, depending on their natural resonance frequencies. This observation suggests that, in addition to increasing reliability of synaptic transmission, bursts of action potentials might provide effective mechanisms for selective communication between neurons. PMID- 12591220 TI - Time windows for effects of protein synthesis inhibitors on Pavlovian conditioning in Hermissenda: behavioral aspects. AB - Inhibitors of protein and mRNA syntheses inhibit long-term memory (LTM), but we lack information about the time windows during which those inhibitors are effective. Anisomycin (a protein synthesis translation inhibitor) was given to Hermissenda crassicornis which had received sufficient Pavlovian conditioning to produce LTM. When tested after 90 min, LTM recall was blocked when anisomycin was administered until 13 min after conditioning and from 65 to 75 min. There was good recall from 16 to 60 min. When tested at 240 min, two periods of sensitivity to anisomycin were revealed: one finished before 20 min, while the other lasted from 60 to 220 min.A brief study of the transcription inhibitor actinomycin-D found its action similar to that of anisomycin, except not inhibiting recall from about 160 to 220 min. PMID- 12591221 TI - Differential effects of emotional arousal in short- and long-term memory in healthy adults. AB - Recent studies demonstrated important differences between short- and long-term memory mechanisms. Besides, the emotional component has a crucial role in memory formation. This study was carried out to answer whether there is a differential influence of emotional arousal in short- and long-term memory in healthy adults. Thirty-one healthy volunteers were divided into two major groups. In the first group long-term memory (LTM) was evaluated, with the testing session one week after training. The second group was tested 1h after training, where short-term memory (STM) was evaluated. Each group was divided in to two subgroups. One half of the volunteers was exposed to an emotionally neutral story, and the other half of each group was exposed to a closely matched but more emotionally arousing story. The testing session consisted of a questionary containing 80 questions of multiple choices. The results were evaluated through percentage of correct answers. Results showed that correct answers were increased, in LTM measures, in the subjects that were given the emotional version of the test. In STM measures, no differences were found between the emotional and neutral version. However, the presentation of emotional story caused an emotional reaction in both groups. The lack of effect of emotional arousal in STM suggests that amygdala is not related to STM mechanisms. Further studies using different approaches are needed to elucidate if STM processes are influenced by emotional arousal. PMID- 12591222 TI - Peripheral 8-OH-DPAT and scopolamine infused into the frontal cortex produce passive avoidance retention impairments in rats. AB - The present study determined whether peripheral injections of the 5HT(1A) agonist (8-OH-DPAT), scopolamine infusions into the frontal cortex, or a combination of both drug treatments would produce impairments in rats trained on passive avoidance. Using a 2x2 design, rats were infused with either bacteriostatic water or 30 microg/1 microl of scopolamine HCl into the frontal cortex 30 min before being trained on passive avoidance. This was followed by injections (ip) of either 0.1% ascorbic acid/bacteriostatic water or 30 microg/kg of 8-OH-DPAT 15 min later. All subjects were tested for retention 72h later. At test, the initial latency to enter into the black shocked compartment and the total time spent in the white safe compartment (TTW) were recorded. Analysis of the latency data indicated that scopolamine and 8-OH-DPAT, when administered singly or in combination, produced amnesia for the task. Assessment of TTW scores, however, revealed that of the three drug-treated groups, only animals treated with 8-OH DPAT alone tended to avoid the previously shocked black compartment and spend more time in the white safe compartment. These data indicate that either stimulating 5-HT(1A) or blocking frontal cortical muscarinic receptors at training impairs passive avoidance performance and that the deficit following the latter treatment is somewhat more extensive. Implications for the role frontal cortical muscarinic and 5HT(1A) receptors play in learning and memory are discussed. PMID- 12591223 TI - Learning associated increase in heat shock cognate 70 mRNA and protein expression. AB - The Morris water maze is a task widely used to investigate cellular and molecular changes associated with spatial learning and memory. This task has both spatial and aversive (swimming related stress) components. It is possible that stress may influence cellular modifications observed after learning the Morris water maze spatial task. Heat shock proteins, also known as stress proteins, are up regulated in response to thermal stress, trauma, or environmental insults. In the rat hippocampus, psychophysiological stress increases the levels of heat shock protein 70 (HSC70). In this study, we investigated whether the expression of the hsc70 gene is modulated in the hippocampus during learning of the Morris water maze task. Five groups of rats were trained in the Morris water maze task for varying amounts of time (either 1, 2, 3, 4, or 5 days). Training consisted of 10 trials/day in which the animals were given 60s to find a submerged platform. Rats were sacrificed 24h after their last training trial. Results showed a significant increase in hsc70 mRNA and protein levels in the hippocampal formation after two and three days of training, respectively. The increase in mRNA and protein was associated with learning but not stress because the increase was not observed in the yoked control animals. These findings suggest that cellular and molecular changes can occur independent of stress. Moreover, the results are the first to implicate hsc70 expression in spatial learning. PMID- 12591224 TI - CS-specific gamma, theta, and alpha EEG activity detected in stimulus generalization following induction of behavioral memory by stimulation of the nucleus basalis. AB - Tone paired with stimulation of the nucleus basalis (NB) induces behavioral memory that is specific to the frequency of the conditioned stimulus (CS), assessed by cardiac and respiration behavior during post-training stimulus generalization testing. This paper focuses on CS-specific spectral and temporal features of conditioned EEG activation. Adult male Sprague-Dawley rats, chronically implanted with a stimulating electrode in the NB and a recording electrode in the ipsilateral auditory cortex, received either tone (6kHz, 70dB, 2s) paired with co-terminating stimulation of the nucleus basalis (0.2s, 100Hz, 80-105 microA, ITI approximately 45s) or unpaired presentation of the stimuli (approximately 200 trials/day for approximately 14 days). CS-specificity was tested 24h post-training by presenting test tones to obtain generalization gradients for the EEG, heart rate, and respiration. Behavioral memory was evident in cardiac and respiratory responses that were maximal to the CS frequency of 6kHz. FFT analyses of tone-elicited changes of power in the delta, theta, alpha, beta1, beta2, and gamma bands in the paired group revealed that conditioned EEG activation (shift from lower to higher frequencies) was differentially spectrally and temporally specific: theta, and alpha to a lesser extent, decreased selectively to 6kHz during and for several seconds following tone presentation while gamma power increased transiently during and after 6kHz. Delta exhibited no CS-specificity and the beta bands showed transient specificity only after several seconds. The unpaired group exhibited neither CS-specific behavioral nor EEG effects. Thus, stimulus generalization tests reveal that conditioned EEG activation is not unitary but rather reflects CS-specificity, with band-selective markers for specific, associative neural processes in learning and memory. PMID- 12591225 TI - Patterns of brain acetylcholine release predict individual differences in preferred learning strategies in rats. AB - Acetylcholine release was measured simultaneously in the hippocampus and dorsal striatum of rats before and during training on a maze that could be learned using either a hippocampus-dependent spatial strategy or a dorsal striatum-dependent turning strategy. A probe trial administered after rats reached a criterion of 9/10 correct responses revealed that about half of the rats used a spatial strategy and half a turning strategy to solve the task. Acetylcholine release in the hippocampus, as well as the ratio of acetylcholine release in the hippocampus vs. the dorsal striatum, measured either before or during training, predicted these individual differences in strategy selection during learning. These findings suggest that differences in release of acetylcholine across brain areas may provide a neurobiological marker of individual differences in selection of the strategies rats use to solve a learning task. PMID- 12591226 TI - The role of cortical cholinergic pre- and post-synaptic receptors in taste memory formation. AB - A number of studies have implicated cholinergic activity in the mediation of learning and memory processes. However, the specific role of muscarinic receptors in memory formation mechanisms is less known. The aim of the present study is to evaluate the effects of muscarinic antagonist M2 presynaptic receptor, AFDX-116 (0.5mM) and M1 and M3 post-synaptic receptor pirenzepine (100mM), as well as a non-selective muscarinic antagonist, scopolamine (136mM), in the insular cortex (IC) during acquisition and retrieval of conditioned taste aversion (CTA). In addition, we evaluate the effects of those antagonists in cortical ACh release by in vivo microdialysis and the effects on the induction of in vivo LTP in the BLA IC projection. The results showed that the cortical microinjections of scopolamine and pirenzepine, but not AFDX-116, produced significant disruption in the acquisition of CTA, without effects during retrieval. Microinjections of scopolamine and AFDX-116 produced significant cortical ACh release, while infusions of pirenzepine did not produce any release. Application of scopolamine and pirenzepine diminished induction of LTP in the BLA-IC projection, but not AFDX-116, as compared with vehicle. The induction of BLA-CI LTP seems to be modulated by post-synaptic muscarinic acetylcholine receptors and not by pre synaptic muscarinic receptors. These results suggest a differential involvement of cholinergic receptors during acquisition and retrieval of aversive memory formation, as well as a differential role of muscarinic receptors in the biochemical and electrophysiological processes that may underlay aversive memory. PMID- 12591227 TI - Epinephrine enhancement of human memory consolidation: interaction with arousal at encoding. AB - Abundant evidence indicates that endogenous stress hormones like epinephrine and cortisol modulate memory consolidation in animals. Despite this evidence, there has been no demonstration that endogenous stress hormones modulate memory consolidation in humans. In the present study, healthy subjects viewed a series of 21 slides, and immediately after received an intravenous infusion of either saline or epinephrine (40 or 80 ng/kg/min). Memory for the first three (primacy) and last three (recency) slides viewed was assessed with an incidental free recall test one week later. Epinephrine dose-dependently increased memory for the primacy slides, but did not affect memory of the recency slides. A subsequent experiment involving new subjects revealed significantly higher electrodermal responses to the primacy compared with recency slides. These findings support the view (Gold & McGaugh, 1975) that endogenous stress hormones modulate memory consolidation for experiences that induce their release. Additionally, they suggest that in humans these hormones may interact with the degree of arousal at initial encoding of information to modulate memory consolidation processes for that information. PMID- 12591228 TI - Decrease in fibrin content of venous thrombi in selectin-deficient mice. AB - The purpose of this study was to quantify the fibrin content of thrombi produced in a mouse model of venous thrombosis and correlate this to thrombus mass. The role of P-selectin, E-selectin, and IL-10 on thrombus fibrin content was analyzed using knockout (KO) mice. Five groups of mice were evaluated: control (N = 10), P selectin KO (N = 7), E-selectin KO (N = 5), combined E-/P-selectin KO (N = 12), and IL-10 KO (N = 10). Venous thrombosis was induced by ligation of the infrarenal IVC. Mice were sacrificed on postoperative days (POD) 2 and 6. Thrombus mass was calculated. Sections of IVC were stained with an antibody that cross reacts with mouse fibrin. The distribution of RGB color pixels was generated from digitized micrographs of the thrombus of each animal. The mean pixel value for each group was compiled and analyzed using 2-way ANOVA. Mean pixel value per group was correlated with the mean thrombus mass per group. Color analysis demonstrated significant decreases in the analyzed fibrin content on POD 2 between the control vs E-/P-selectin KO (P < 0.05) and control vs IL-10 KO (P < 0.05) groups. In addition, significantly less fibrin staining was noted on POD-6 between the control vs E-selectin KO (P = 0.03), control vs P-selectin KO (P = 0.01), and control vs E-/P-selectin KO (P < 0.01). There was a strong overall correlation between the mean pixel value for each group and the thrombus mass (R = 0.964; P < 0.01). This study demonstrates a difference in fibrin content of thrombi produced in animals deficient in E-selectin, P-selectin, and IL-10, supporting their importance in thrombus amplification, fibrin formation, and the mass of thrombus formed. PMID- 12591229 TI - Novel protection strategy for pulmonary transplantation. AB - BACKGROUND: Ischemia-reperfusion injury continues to represent a significant challenge to successful lung transplantation. Traditional pulmonary ischemic protection is performed using hypothermic hyperkalemic depolarizing solutions to reduce the metabolic demands of the ischemic organ. Measures to further reduce the effects of ischemic injury have focused on the reperfusion period. We tested the hypothesis that novel physiologic hyperpolarizing solutions-using ATP dependent potassium channel (K(ATP)) openers-given at the induction of ischemia, will reduce cellular injury and provide superior graft function even after prolonged periods of ischemia. METHODS: An isolated blood-perfused ventilated rabbit lung model was used to study lung injury. Airway, left atrial, and pulmonary artery pressures were measured continuously during the 2-h reperfusion period. Oxygenation, as a surrogate of graft function, was measured using intermittent blood gas analysis of paired left atrial and pulmonary artery blood samples. Graft function was measured by oxygen challenge technique (F(i)O(2) = 1.0). Wet-to-dry ratio was measured at the conclusion of the 2-h reperfusion period. Control (Group I) lungs were perfused with modified Euro-Collins solution (depolarizing) and reperfused immediately (no ischemia). Traditional protection lungs were perfused with modified Euro-Collins flush solution and stored for 4 h (Group II) or 18 h (Group III) at 4 degrees C before reperfusion. Novel protection (Group IV) lungs were protected with a hyperpolarizing solution containing 100 nM Aprikalim, a specific K(ATP) channel opener, added to the modified Euro-Collins flush solution and underwent 18 h of ischemic storage at 4 degrees C before reperfusion. RESULTS: Profound graft failure was measured after 18 h of ischemic storage with traditional protection strategies (Group III). Graft function was preserved by protection with hyperpolarizing solutions even for prolonged ischemic periods (Group IV). Wet-to-dry weight ratio, airway, left atrial, and pulmonary artery pressures were not significantly different between the groups. CONCLUSIONS: We have created a model of predictable lung injury. Membrane hyperpolarization with a K(ATP) channel opener (PCO) provides superior prolonged protection from ischemia-reperfusion injury in an in vitro model of pulmonary transplantation. PMID- 12591230 TI - Vascular endothelial growth factor increases the migration and proliferation of smooth muscle cells through the mediation of growth factors released by endothelial cells. AB - BACKGROUND: Vascular endothelial growth factor (VEGF), a highly specific chemotactic and mitogenic factor for vascular endothelial cells (EC), appears to be involved in the development of atherosclerosis. The purpose of our study was to assess if VEGF might indirectly stimulate SMC migration and proliferation in a EC-SMC coculture system, through the mediation of growth factors released by EC. METHODS: Bovine aortic SMC were cocultured with bovine aortic EC treated with hrVEGF, to assess SMC proliferation and migration. The release and mRNA expression of basic fibroblast growth factor (bFGF) and transforming growth factor beta(1) (TGFbeta(1)) were assessed by ELISA and PCR analysis. RESULTS: hrVEGF (10 ng/ml), added to EC cocultured with SMC, induced a significant increase in tritiated thymidine uptake by SMC as compared to controls (P < 0.01) and a significant increase in SMC migration in respect to control (27%; P < 0.01). EC stimulated with hrVEGF increased the release and the expression of bFGF and decreased the release and the expression of TGFbeta(1) with a statistically significant difference in respect to controls (P < 0.001). CONCLUSIONS: VEGF indirectly stimulates SMC proliferation and migration through the modulation of bFGF and TGFbeta(1) released by EC. PMID- 12591231 TI - Clinically relevant thermal preconditioning attenuates ischemia-reperfusion injury. AB - INTRODUCTION: Thermal preconditioning has previously been shown to attenuate ischemia-reperfusion induced injuries, possible due to increased expression of heat shock proteins (HSP). The model of thermal preconditioning used, however, was not clinically relevant as preconditioning was to 41 degrees C, leading to cellular damage. Our aim was thus to establish a novel and clinically applicable method of preconditioning. MATERIALS AND METHODS: Twenty-six male Sprague-Dawley rats were split into three groups (nine control, nine ischemia-reperfusion, and eight preconditioned followed by ischemia-reperfusion). To precondition the animals, they were anesthetized and, using a water bath, their core temperature was raised by 1 degrees C for 15 min once a day for five successive days. I/R injury consisted of 30 min of aortic cross-clamping followed by 120 min of reperfusion; control animals had a laparotomy only. Indicators of lung injury were tissue myeloperoxidase, broncho-alveolar lavage protein concentration, and tissue edema. Tissue heat shock protein expression was detected by Western blot analysis. RESULTS: Lower torso ischemia-reperfusion causes significant lung injury versus control, with raised levels of myeloperoxidase 4.53 iu/g to 7.88 iu/g (P < 0.05), raised B.A.L. protein concentration 419 microg/ml to 684 microg/ml (P < 0.05) and altered wet dry ratio 4.63 to 5.50. Clinically relevant thermal preconditioning attenuates all of these parameters back to control levels: myeloperoxidase 3.87 iu/g (P < 0.05 vs I/R), B.A.L. to 284 microg/ml (P < 0.01 vs I/R) and wet dry ratio to 4.44 (P < 0.05 vs I/R). Western blot demonstrated increased expression of H.S.P. 72 in the preconditioned group versus control and I/R alone. Western blot demonstrated increased expression of HSP72 in the preconditioned group vs control and I/R alone. CONCLUSIONS: We conclude that clinically applicable thermal preconditioning can attenuate ischemia-reperfusion induced lung injury, possibly through increased expression of HSP72. PMID- 12591232 TI - Effect of ileocolonic transposition on gut morphology, gene expression, and function. AB - Intestinal morphology and expression of brush border enzymes can be dramatically altered by nutrient composition and position along the longitudinal gut axis. In contrast, endocrine gene expression appears to be relatively "imprinted" and is not significantly altered by positional changes. The purpose of our study was to assess the effects of transposing the ileum into the colonic position on intestinal morphology, gene expression, and enzyme activity. For this study, male Sprague-Dawley rats underwent either transposition of an approximately 8-cm segment of distal ileum into the colonic position or sham operation. Rats were sacrificed 2 months after operation and transposed or sham ileum was assessed. We found that transposition of the ileum to the colonic position resulted in no change in the ileal mucosal architecture. Furthermore, no change in the expression of the endocrine genes neurotensin and PYY or the apoptotic genes was detected. Although sucrase-isomaltase activity was decreased in the transposed ileum, this decrease did not achieve statistical significance. These results demonstrate that the ileum can be successfully interposed into the colonic position. In this location, the ileum functions normally without changes in the villus morphology or gene expression pattern despite the change in position and composition of the luminal content. PMID- 12591233 TI - Polymorphonuclear neutrophils promote rFGF-2-induced angiogenesis in vivo. AB - BACKGROUND: The role of neutrophils in angiogenesis remains largely unknown. Recent evidence has shown that polymorphonuclear neutrophils (PMNs) produce several proangiogenic cytokines, including VEGF, TNF-alpha, IL-1, IL-6, and IL-8. In addition, PMN-derived proteinases promote endothelial cell migration. We hypothesized that PMNs may facilitate angiogenesis and that reducing circulating PMNs might alter the host angiogenic response. MATERIALS AND METHODS: We utilized a corneal pocket assay to compare rFGF-2-induced vessel formation in the corneas of mice with normal levels of circulating neutrophils to those in a neutropenic state. Circulating PMNs were reduced using serial intraperitoneal injections of monoclonal antibody to Gr-1. Slow release rFGF2 pellets were implanted into the corneas of neutropenic mice and controls. Corneal neovascularization, measured as vessel length and area of vessel in-growth, was quantified using slit-lamp microscopy on day 7. RESULTS: The average number of circulating PMNs was significantly reduced in the experimental group compared to the control group on days 1-7 (P < 0.05). No statistical differences in circulating monocytes or lymphocytes were observed from days 0 to 6. Mice in the experimental group had a vascular area of 2.58 +/- 0.2 mm(2) compared to 3.55 +/- 0.3 mm(2) in the control group (P < 0.05). CONCLUSIONS: Corneal neovascularization in response to rFGF-2 is diminished by PMN depletion. PMNs play an important role in facilitating rFGF 2-induced angiogenesis. PMID- 12591234 TI - TGF-beta stimulates collagen (I) in vascular smooth muscle cells via a short element in the proximal collagen promoter. AB - BACKGROUND: Accumulation of extracellular matrix contributes to the development of intimal hyperplasia. Transforming growth factor beta (TGF-beta) stimulates the production of several matrix proteins in vascular smooth muscle cells (SMC) including type I collagen, but the underlying mechanisms of TGF-beta's effects are not well understood. MATERIALS AND METHODS: The effect of TGF-beta on type I collagen biosynthesis was determined by a [3H]proline incorporation assay and Northern blotting. The promoter of human alpha2(I) procollagen (COL1A2) gene was analyzed by transient transfection analysis and gel mobility shift assay. RESULTS: Treatment of human vascular SMC with TGF-beta stimulated collagen synthesis and increased the level of alpha2(I) collagen mRNA. A collagen luciferase reporter gene, constructed by linking the human COL1A2 promoter with the firefly luciferase gene, was transiently expressed in human SMC. Treatment with TGF-beta significantly stimulated the activity of this collagen-luciferase reporter. Using deletion analysis, we identified a 150 bp DNA fragment (-334 to 184) in the human COL1A2 promoter as the site through which TGF-beta mediates collagen gene expression in human SMC. Gel mobility shift assays demonstrated that this 150 bp DNA fragment formed conjugates with multiple nuclear factors derived from SMC, a process that was further enhanced by TGF-beta. CONCLUSIONS: TGF-beta stimulates the human type I collagen gene via a DNA element located in the proximal region of its promoter. Interventions that disrupt interaction between this DNA element and nuclear factors may block the production of collagen in response to TGF-beta and consequently may have a significant effect on the development of intimal hyperplasia. PMID- 12591235 TI - Mesenteric lymph duct ligation prevents shock-induced RBC deformability and shape changes. AB - OBJECTIVES: The exact mechanisms that lead to RBC deformability and shape changes after trauma/hemorrhagic shock remain unknown. We hypothesize that RBC injury is caused in part by gut injury and is mediated by gut-derived factors carried in the intestinal lymph. MATERIALS AND METHODS: RBC deformability was measured by a laser-assisted ektacytometer before and after trauma/hemorrhagic shock (T/HS) in 6 rats whose mesenteric lymph duct had been ligated and in 10 rats subjected to T/HS without duct ligation. In this assay a decrease in the elongation index is a marker of decreased RBC deformability. RBC shape was examined by scanning electron microscopy. RESULTS: In the T/HS rats, the RBC elongation index decreased after T/HS from a preshock value of 0.064 +/- 0.011 to 0.052 +/- 0.009 (P < 0.01) and remained low (0.049 +/- 0.010) even at 3 h after resuscitation. In contrast, the elongation index did not decrease after T/HS in the lymph duct ligated rats (0.062 +/- 0.004 vs. 0.056 +/- 0.005, P = NS). Likewise, the T/HS rats, but not the duct-ligated T/HS rats, had a significant increase in the percentage of abnormally shaped RBCs when studied by electron microscopy. CONCLUSIONS: Interruption of lymph flow from the gut into the bloodstream by lymph duct ligation prevents T/HS-induced RBC damage. Because decreased RBC deformability contributes to impaired perfusion of the microcirculation, preservation of RBC deformability may decrease the incidence of T/HS-induced organ dysfunction. PMID- 12591236 TI - Cyclic AMP inhibits production of interleukin-6 and migration in human vascular smooth muscle cells. AB - BACKGROUND: Gene expression induced by tumor necrosis factor-alpha (TNF-alpha) is involved in the regulation of vascular smooth muscle cell (VSMC) proliferation and migration, two events critical to formation of stenotic vascular lesions. In some systems, elevating adenosine 3',5'-cyclic monophosphate (cyclic AMP) inhibits TNF-alpha induced gene transcription. We recently demonstrated that interleukin-6 (IL-6) was chemotactic to VSMC. Therefore, we tested the hypothesis that elevating cyclic AMP would inhibit TNF-alpha-mediated IL-6 expression and VSMC migration. MATERIALS AND METHODS: VSMC were cultured from saphenous vein remaining after coronary artery bypass grafting. Migration of VSMC through a porous membrane was determined. Intracellular cyclic AMP was elevated by exposing the cells to forskolin or 8-Br-cyclic AMP and was measured by radioimmunoassay. IL-6 was measured by enzyme-linked immunosorbent assay. RESULTS: TNF-alpha induced migration of VSMC in a concentration-dependent manner. Incubation of cells with forskolin significantly increased cyclic AMP. Co-incubation of cells with TNF-alpha in combination with 8-Br-cyclic AMP or forskolin inhibited migration by approximately 25 and 70%, respectively. Incubation with TNF-alpha increased release of IL-6 from VSMC 18-fold over basal. This stimulated release was inhibited by either 8-Br-cyclic AMP or forskolin. In cells stimulated with TNF-alpha, addition of an antibody to IL-6 reduced migration by 25%. CONCLUSIONS: These data show that IL-6 produced by VSMC contributes to cell migration induced by TNF-alpha. Further, elevating cyclic AMP inhibited TNF-alpha-induced release of IL-6, and migration of VSMC. These results are consistent with the notion that mechanisms that increase intracellular cyclic AMP, such as activation of beta adrenergic receptors on VSMC, act as a brake on cell migration. PMID- 12591237 TI - Limitations of ischemic tolerance in oxidative skeletal muscle: perfusion vs tissue protection. AB - OBJECTIVES: This study determined if ischemic tolerance occurs in oxidative skeletal muscle following a severe ischemia/reperfusion (I/R) insult and if such protection involves the induction of nitric oxide synthase (NOS). METHODS: The soleus muscle of male Wistar rats (250-350 g) was preconditioned (PC + I/R) using five cycles of ischemia (10 min) and reperfusion (10 min) or had no PC (I/R) and 24 h later 2 h no-flow ischemia was induced. Calcium dependent (cNOS) and independent (iNOS) NOS activities were determined from PC (n = 5), or sham (n = 5) and the role of iNOS was tested by application of aminoguanidine (AMG) (100 microM; n = 4) to the muscle bath. Direct measures of the number of perfused capillaries (Npc; #/mm) during 90-min reperfusion were obtained using intravital microscopy. Tissue injury was estimated using the fluorescent vital dyes ethidium bromide (E; labels injured cells) and bisbenzimide (B; labels all cells) and expressed as the ratio E/B. RESULTS: PC prevented microvascular flow deficits (Npc:I/R = 23.4 +/- 1.3 vs PC + I/R = 29.9 +/- 1.1) and resulted in a modest, but significant reduction (21%) in tissue injury (I/R = 0.82 +/- 0.03 vs PC + I/R = 0.64 +/- 0.04). PC led to a nine fold increase in iNOS activity, but decreased cNOS activity by 94% compared to sham. AMG prevented the parenchymal protection following PC, but had no effect on microvascular perfusion. CONCLUSIONS: Ischemic tolerance, 24 h following PC, preserved microvascular perfusion, but only modestly improved tissue viability in the soleus muscle. PMID- 12591238 TI - Maturation-associated increase in IP3 receptor type 1: role in conferring increased IP3 sensitivity and Ca2+ oscillatory behavior in mouse eggs. AB - Maturation of mouse oocytes is accompanied by an increase in sensitivity to inositol 1,4,5-trisphosphate (IP(3))-mediated release of intracellular calcium. To test the hypothesis that the maturation-associated 1.5- to 2.0-fold increase in the mass of the type 1 IP(3) receptor (IP(3)R-1) confers this increase in IP(3) sensitivity, we employed RNA interference to prevent this change in IP(3)R 1 protein level. Microinjection into germinal vesicle (GV)-intact oocytes of dsRNA corresponding to the IP(3)R-1 sequence resulted in a >90% reduction in the amount of maternal IP(3)R-1 mRNA and prevented the maturation-associated increase in the mass of the IP(3)R-1 protein. These injected oocytes matured to metaphase II, and there was no effect on the maturation-associated increases in p34(cdc2)/cyclin B kinase and MAP kinase activities or the global pattern of protein synthesis. IP(3)-induced cortical granule exocytosis was significantly decreased in these eggs when compared with controls previously injected with enhanced green fluorescent protein (EGFP) dsRNA. Following insemination, the IP(3)R-1 dsRNA-injected eggs displayed significantly fewer Ca(2+) transients than controls, and the duration of the first Ca(2+) transient was about half that of controls. These results support the hypothesis that the maturation-associated increase in the mass of IP(3)R-1 confers the increase in IP(3)-sensitivity that is observed following oocyte maturation and is necessary for the proper Ca(2+) oscillatory pattern following insemination. PMID- 12591239 TI - Wnt1 and wnt10b function redundantly at the zebrafish midbrain-hindbrain boundary. AB - Wnt signals have been shown to be involved in multiple steps of vertebrate neural patterning, yet the relative contributions of individual Wnts to the process of brain regionalization is poorly understood. Wnt1 has been shown in the mouse to be required for the formation of the midbrain and the anterior hindbrain, but this function of wnt1 has not been explored in other model systems. Further, wnt1 is part of a Wnt cluster conserved in all vertebrates comprising wnt1 and wnt10b, yet the function of wnt10b during embryogenesis has not been explored. Here, we report that in zebrafish wnt10b is expressed in a pattern overlapping extensively with that of wnt1. We have generated a deficiency allele for these closely linked loci and performed morpholino antisense oligo knockdown to show that wnt1 and wnt10b provide partially redundant functions in the formation of the midbrain hindbrain boundary (MHB). When both loci are deleted, the expression of pax2.1, en2, and her5 is lost in the ventral portion of the MHB beginning at the 8-somite stage. However, wnt1 and wnt10b are not required for the maintenance of fgf8, en3, wnt8b, or wnt3a expression. Embryos homozygous for the wnt1-wnt10b deficiency display a mild MHB phenotype, but are sensitized to reductions in either Pax2.1 or Fgf8; that is, in combination with mutant alleles of either of these loci, the morphological MHB is lost. Thus, wnt1 and wnt10b are required to maintain threshold levels of Pax2.1 and Fgf8 at the MHB. PMID- 12591240 TI - Microarray analysis of knockout mice identifies cyclin D2 as a possible mediator for the action of thyroid hormone during the postnatal development of the cerebellum. AB - Thyroid hormone is a major regulator of postnatal brain development, but the precise molecular mechanisms underlying its action in this organ remain poorly understood. We used microarray analysis to identify new target genes in brain. Thyroid hormone treatment of hypothyroid Pax8(-/-) knockout mice, which lack thyroid follicular cells, had a very limited global effect on brain transcripts. This analysis mainly identified cyclin D2 as a new thyroid hormone target gene in the cerebellum of hypothyroid mice. Thyroid hormone receptor (TRalpha and/or TRbeta) knockout mice studies provided further genetic evidence that cyclin D2 is likely to mediate the antiapoptotic effect exerted by thyroid hormone on the cerebellum external granular layer neuroblasts but that this transcriptional activation is not directly exerted by the thyroid hormone receptors. PMID- 12591242 TI - The pleiotropic mutation dar1 affects plant architecture in Arabidopsis thaliana. AB - Shoot architecture is shaped upon the organogenic activity of the shoot apical meristem (SAM). Such an activity relies on the balance between the maintenance of a population of undifferentiated cells in the centre of the SAM and the recruitment of organ founder cells at the periphery. A novel mutation in Arabidopsis thaliana, distorted architecture1 (dar1), is characterised by disturbed phyllotaxy of the inflorescence and consumption of the apical meristem late in development. SEM and light microscopy analyses of the dar1 SAM reveal an abnormal partitioning of meristematic domains, and mutations known to affect the SAM structure and function were found to interact with dar1. Moreover, the mutant shows an alteration of the root apical meristem (RAM) structure. Those observations support the hypothesis that DAR1 has a role in meristem maintenance and it is required for the normal development of Arabidopsis inflorescence during plant life. PMID- 12591241 TI - A tissue restricted role for the Xenopus Jun N-terminal kinase kinase kinase MLK2 in cement gland and pronephric tubule differentiation. AB - The MLK family of mitogen activated protein kinase kinase kinases (MAPKKK) has been shown to activate Jun N-terminal kinase/stress-activated protein kinase 1 (JNK/SAPK1). However, little is known of the in vivo functions of the MLKs. We have identified a Xenopus laevis MLK that shows highest homology with mammalian MLK2 (62%) and, like MLK2, interacts preferentially with the Rho-family GTPase Rac. xMLK2 was expressed zygotically from late gastrula/early neurula. Surprisingly, this expression was restricted to the cement gland, the brain, and the pronephros. In the differentiating cement gland, xMLK2 expression correlated with cell elongation and the onset of a previously unobserved apoptotic phase, while in the pronephros, expression corresponded with the differentiation and opening of the nephric tubules. Overexpression of xMLK2 in COS7 cells led to a SEK1/MKK4 (MAPKK)-dependent hyperactivation of JNK in response to UV irradiation. xMLK2 was shown to be required for normal cement gland development and pronephric tubule formation using antisense inactivation and a dominant negative xMLK2. The data suggest a novel role for the MLKs as tissue-restricted mediators of signal transduction. They also suggest that tissue-specific responses to common extracellular signals may in part result from the programmed expression of MAPKKKs with differing specificities. PMID- 12591244 TI - The drosophila angiotensin-converting enzyme homologue Ance is required for spermiogenesis. AB - The Angiotensin-converting enzyme (Ance) gene of Drosophila melanogaster is a homologue of mammalian angiotensin-converting enzyme (ACE), a peptidyl dipeptidase implicated in regulation of blood pressure and male fertility. In Drosophila, Ance protein is present in vesicular structures within spermatocytes and immature spermatids. It is also present within the lumen of the testis and the waste bag, and is associated with the surface of elongated spermatid bundles. Ance mRNA is found mainly in large primary spermatocytes and is not detectable in cyst cells. Testes lacking germ cells have reduced levels of ACE activity, and no Ance protein is detectable by immunocytochemistry, indicating that the germ cells are the major site of Ance synthesis. Ance mutant testes lack individualised sperm and have very few actin-based individualisation complexes. Spermatid nuclei undergo scattering along the cyst and have abnormal morphology, similar to other individualisation mutants. Mutant spermatids also have abnormal ultrastructure with grossly defective mitochondrial derivatives. The failure of Ance mutant testes to form individualisation complexes may be due to a failure in correct spermatid differentiation. Taken together, the expression pattern and mutant phenotype suggest that Ance is required for spermatid differentiation, probably through the processing of a regulatory peptide synthesised within the developing cyst. PMID- 12591243 TI - None of the integrins known to be present on the mouse egg or to be ADAM receptors are essential for sperm-egg binding and fusion. AB - Antibody inhibition and alpha6beta1 ligand binding experiments indicate that the egg integrin alpha6beta1 functions as a receptor for sperm during gamete fusion; yet, eggs null for the alpha6 integrin exhibit normal fertilization. Alternative integrins may be involved in sperm-egg binding and fusion and could compensate for the absence of alpha6beta1. Various beta1 integrins and alphav integrins are present on mouse eggs. Some of these integrins are also reported to be receptors for ADAMs, which are expressed on sperm. Using alpha3 integrin null eggs, we found that the alpha3beta1 integrin was not essential for sperm-egg binding and fusion. Oocyte-specific, beta1 integrin conditional knockout mice allowed us to obtain mature eggs lacking all beta1 integrins. We found that the beta1 integrin null eggs were fully functional in fertilization both in vivo and in vitro. Furthermore, neither anti-mouse beta3 integrin function-blocking monoclonal antibody (mAb) nor alphav integrin function-blocking mAb inhibited sperm binding to or fusion with beta1 integrin null eggs. Thus, function of beta3 or alphav integrins does not seem to be involved in compensating for the absence of beta1 integrins. These results indicate that none of the integrins known to be present on mouse eggs or to be ADAM receptors are essential for sperm-egg binding/fusion, and thus, egg integrins may not play the role in gamete fusion previously attributed to them. PMID- 12591245 TI - The protocadherin papc is involved in the organization of the epithelium along the segmental border during mouse somitogenesis. AB - The anterior and posterior halves of individual somites adopt distinct fates during somitogenesis, which is crucial for establishing the metameric pattern of axial tissues such as the vertebral column and peripheral nerves. Genetic analyses have demonstrated that the specification of cells to an anterior or posterior fate is intimately related to the process of segmentation. Inactivation of the transcription factor Mesp2, or components of the Notch signaling pathway, led to defects in segmentation and a loss of anterior/posterior polarity. Target genes in mice that could mediate the morphological events associated with segmentation or polarity have not been identified. Studies in Xenopus and zebrafish have demonstrated that the protocadherin, papc, is expressed in an anterior-specific manner in the presumptive somites of the presomitic mesoderm and is required for normal somitogenesis. Here, we examine the role of papc in directing segmentation in the mouse. We demonstrate that papc is expressed in a dynamic pattern within the first two presumptive somites (0 and -1) at the anterior end of the presomitic mesoderm. The domain of papc transcription in somite 0 starts broad and becomes progressively restricted to the anterior edge. Transcription in somite -1 over the same time remains broad. Analysis of targeted null mutations revealed that transcription of papc is dependent on Mesp2. The dynamic nature of papc transcription in somite 0 requires the expression of lunatic fringe, which modifies the activation of the Notch signaling pathway and is required for proper segmentation of somites. Treatment of embryonic mouse tails in a hanging drop culture with a putative dominant-negative mutation of papc disrupted the epithelial organization of cells at the segmental borders between somites. Together, these data indicate that papc is an important regulator of somite epithelialization associated with segmentation. PMID- 12591246 TI - Pbx1 regulates nephrogenesis and ureteric branching in the developing kidney. AB - Pbx1 encodes a TALE homeodomain transcription factor that regulates developmental gene expression in a variety of tissues. Loss-of-function studies have demonstrated a critical role for Pbx1 in cellular proliferation and patterning and suggest its involvement in numerous regulatory pathways. In this study, examination of metanephric development in Pbx1(-/-) embryos was conducted to further elucidate Pbx1-dependent processes during organogenesis. Prior to death at E15.5, Pbx1(-/-) embryos displayed kidneys that were reduced in size, axially mispositioned, and in more severe cases, exhibited unilateral agenesis. Analysis with molecular markers revealed the effective induction of tubulogenic mesenchyme; however, Pbx1(-/-) kidneys contained fewer nephrons and were characterized by expanded regions of mesenchymal condensates in the nephrogenic zone. Despite the restricted expression of Pbx1 in metanephric mesenchyme, developing nephrons, and stroma, decreased branching and elongation of the ureter were also observed. Moreover, heterologous recombination studies with explant cultures verified that Pbx1(-/-) renal defects arose exclusively from mesenchymal dysfunction. Taken together, these data establish a role for Pbx1 in mesenchymal epithelial signaling and demonstrate that Pbx1 is an essential regulator of mesenchymal function during renal morphogenesis. PMID- 12591247 TI - Temporally dynamic response to Wingless directs the sequential elaboration of the proximodistal axis of the Drosophila wing. AB - The Drosophila wing imaginal disc gives rise to three main regions along the proximodistal axis of the dorsal mesothoracic segment: the notum, proximal wing, and wing blade. Development of the wing blade requires the Notch and wingless signalling pathways to activate vestigial at the dorsoventral boundary. However, in the proximal wing, Wingless activates a different subset of genes, e.g., homothorax. This raises the question of how the downstream response to Wingless signalling differentiates between proximal and distal fate specification. Here, we show that a temporally dynamic response to Wingless signalling sequentially elaborates the proximodistal axis. In the second instar, Wingless activates genes involved in proximal wing development; later in the third instar, Wingless acts to direct the differentiation of the distal wing blade. The expression of a novel marker for proximal wing fate, zfh-2, is initially activated by Wingless throughout the "wing primordium," but later is repressed by the activity of Vestigial and Nubbin, which together define a more distal domain. Thus, activation of a distal developmental program is antagonistic to previously established proximal fate. In addition, Wingless is required early to establish proximal fate, but later when Wingless activates distal differentiation, development of proximal fate becomes independent of Wingless signalling. Since P element insertions in the zfh-2 gene result in a revertable proximal wing deletion phenotype, it appears that zfh-2 activity is required for correct proximal wing development. Our data are consistent with a model in which Wingless first establishes a proximal appendage fate over notum, then the downstream response changes to direct the differentiation of a more distal fate over proximal. Thus, the proximodistal domains are patterned in sequence and show a distal dominance. PMID- 12591248 TI - Differential non-target-derived repulsive signals play a critical role in shaping initial axonal growth of dorsal root ganglion neurons. AB - Initial trajectories of dorsal root ganglion (DRG) axons are shaped by chemorepulsive signals from surrounding tissues. Although we have previously shown that axonin-1/SC2 expression on DRG axons is required to mediate a notochord-derived chemorepulsive signal, Dev. Biol. 224, 112-121), other molecules involved in the non-target-derived repulsive signals are largely unknown. Using coculture assays composed of tissues derived from the chick embryo or mutant mice treated with function-blocking antibodies and phosphatidylinositol specific phospholipase C, we report here that the chemorepellent semaphorin 3A (Sema3A) and its receptor neuropilin-1 are required for mediating the dermamyotome- and notochord-derived, but not the ventral spinal cord-derived, chemorepulsive signal for DRG axons. The dermamyotome-derived chemorepulsion is exclusively dependent on Sema3A/neuropilin-1, whereas other molecules are also involved in the notochord-derived chemorepulsion. Chemorepulsion from the ventral spinal cord does not depend on Sema3A/neuropilin-1 but requires axonin-1/SC2 to repel DRG axons. Thus, differential chemorepulsive signals help shape the initial trajectories of DRG axons and are critical for the proper wiring of the nervous system. PMID- 12591249 TI - Key factors affecting spatial variation of methane emissions from freshwater marshes. AB - To understand the mechanism for spatial variation of CH(4) emissions from marshes grown with different type of plants in a region and plots within a certain marsh grown with one type of plants, we measured CH(4) emissions from a region in which eutrophic freshwater marshes were divided into three types: Carex lasiocarpa, Carex meyeruana and Deyeuxia angustifolia according to plant type as well as CH(4) concentration in porewater, aboveground plant biomass and stem density in situ in Sanjiang Plain of Northeast China in August 2001. Spatial variation of CH(4) emissions from both different marshes in a region and different plots within a certain marsh was high. The flux rates of CH(4) emissions from three marshes ranged from 17.2 to 66.5 mg CH(4) m(-2)h(-1) with 34.76% of variation coefficient, whereas the values in Carex lasiocarpa, Carex meyeriana and Deyeuxia angustifolia marshes varied from 21.6 to 66.5 (39.61%), from 17.2 to 45.0 (29.26%) and from 19.1 to 33.0 mg CH(4) m(-2)h(-1) (17.51%), respectively. Both the flux rates and spatial variation of CH(4) emissions strongly increased as standing water depth increased significantly. Standing water depth greatly governed the spatial variation of CH(4) emissions from different marshes in a region by changing the amount of plant litters inundated in standing water, which provided labile organic C for methanogens and controlled CH(4) concentrations in porewater. Moreover, the aboveground plant biomass determined spatial variation of CH(4) emissions from plots within a certain marsh by controlling the pathways (stem density) of CH(4) emissions from the marsh into the atmosphere. PMID- 12591250 TI - Light-induced formation of hydroxyl radicals in fog waters determined by an authentic fog constituent, hydroxymethanesulfonate. AB - The determination of the photo-production rate of hydroxyl radical (OH) in atmospheric liquids is of fundamental importance to an understanding of atmospheric aquatic chemistry. Recently, several studies have been performed to examine the photo-chemical formation rate of OH in cloud and fog waters using a free radical quenching technique with addition of a relatively large concentration of organic compounds as an OH scavenger. The addition of free radical scavenger chemicals may significantly alter the nature of the sample water and its OH production rate. In this paper, an authentic constituent, hydroxymethanesulfonate, is proposed as a free radical probe for the measurement of photo-chemical generation rate of OH in fog water. At 313 nm, an apparent quantum yield for the production of OH in a fog water was found to be 0.012+/ 0.001, indicating that aqueous-phase photo-chemical processes could represent a significant and may be a dominant source of OH in atmospheric liquids. PMID- 12591251 TI - Methane and nitrous oxide emissions from an irrigated rice of North India. AB - Upland rice was grown in the kharif season (June-September) under irrigated condition in New Delhi, India (28 degree 40'N and 77 degree 12'E) to monitor CH4 and N2O emission, as influenced by fertilizer urea, ammonium sulphate and potassium nitrate alone (at 120 kg ha-1) and mixed with dicyandiamide (DCD), added at 10% of applied N. The experimental soil was a typic ustochrept (Inceptisol), clay loam, in which rice (Oryza sativa L., var. Pusa-169, duration: 120-125 days) was grown and CH4 and N2O was monitored for 105 days by closed chamber method, starting from the 5 days and 1 day after transplanting, respectively. Methane fluxes had a considerable temporal variation (CV=52-77%) and ranged from 0.05 (ammonium sulphate) to 3.77 mg m-2 h-1 (urea). There was a significant increase in the CH4 emission on the application of fertilizers while addition of DCD with fertilizers reduced emissions. Total CH4 emission (105 days) ranged from 24.5 to 37.2 kg ha-1. Nitrous oxide fluxes were much lower than CH4 fluxes and had ranged from 0.18 to 100.5 g m-2 h-1 with very high temporal variation (CV=69-143%). Total seasonal N2O emission from different treatments ranged from 0.037 to 0.186 kg ha-1 which was a N loss of 0.10-0.12% of applied N. All the fertilizers significantly increased seasonal N2O emission while application of DCD reduced N2O emissions significantly in the range of 10-53%. PMID- 12591252 TI - Effects of ultraviolet-B radiation on behaviour and growth of three species of amphibian larvae. AB - Effects of ultraviolet-B (UV-B) radiation on amphibian embryos have been investigated in a number of studies, but the effects on larvae have received less attention. We investigated the effects of UV-B radiation on the behaviour and growth of larvae of three amphibians (Rana arvalis, Rana temporaria and Bufo bufo) in two different experiments. First, we tested whether larvae of the three species actively avoid UV-B exposure if given a choice. We found no evidence for active avoidance of UV-B or changes in activity in the presence of UV-B in any of the species. Second, we assessed the effects of natural (1.25 kJm(-2)) and enhanced (1.58 kJm(-2)) UV-B radiation on the survival and growth of the three species and found that the exposure to UV-B radiation did not have any effect on survival rates of any of the species. However, UV-B radiation had a positive effect on the growth of R. arvalis and R. temporaria, whereas the growth of B. bufo tadpoles was unaffected by the UV-B treatments. Our results suggest that a short-term exposure to UV-B radiation does not induce any UV-B avoidance behaviour in tadpoles of these three species. Furthermore, unlike some previous studies, the results suggest that the young tadpoles of these species are not negatively affected by UV-B radiation. In fact, our results demonstrate that a moderate amount of UV-B radiation enhance tadpole growth rates in two of the three species. PMID- 12591253 TI - Spectral attenuation of solar ultraviolet radiation in humic lakes in Central Finland. AB - The attenuation of solar ultraviolet (UV) radiation in five lakes in Central Finland was evaluated through field measurements and/or by determining the optical properties of the lake water during summer 1999. Spectral UV irradiance in the air and at several depths underwater was measured in three lakes (Lake Palosjarvi, Konnevesi, and Jyvasjarvi) with dissolved organic carbon (DOC) ranging from 4.9 to 8.7 mg l(-1) and chlorophyll a ranging from 1.6 to 16 g l( 1). According to the field measurements, 99% of the UV-B radiation was attenuated in approximately a half meter water column in the clearest lake. In the UV-A region at 380 nm, the corresponding attenuation occurred in the upper one meter. In a small humic lake (DOC 13.2-14.9 mg l(-1)) UV-B radiation was attenuated to 1% of the subsurface irradiance within the top 10 cm water column, whereas UV-A radiation (at 380 nm) penetrated more than twice as deeply (maximum 25 cm), as predicted from the absorption coefficients. These results suggest the importance of the dissolved fraction of lake water in governing the UV attenuation in lakes. This was seen from the significant relationship between the vertical attenuation coefficients (Kd) based on field measurements and the absorption coefficients (ad) derived from spectrophotometric laboratory scannings, as well as between Kd and DOC. PMID- 12591254 TI - Effect of global climate change and human disturbances on tree diversity of the forest regenerating from clear-cuts of mixed broadleaved Korean pine forest in Northeast China. AB - Studies on the combined effects of global climate change and human disturbances are important for biodiversity conservation and natural resources management. Here we use the modified forest dynamics model to simulate the tree diversity change of a typical mixed broadleaved Korean pine forest regenerating from clear cuts in Northeast China in response to global climate change, double concentration of CO(2) and human disturbances during the next 50 years. We consider the following climate change scenario: the annual temperature will increase 2 degrees C, the annual precipitation will increase 10% and CO(2) concentration will increase to 700 microll(-1) linearly in 50 years. Five kinds of human disturbances under climate change are considered: logging which removes all trees with diameter at the breast height of more than 50 cm; removing all individuals of any one species; and removing all individuals of shade tolerant, shade intolerant and medium type tree species, respectively. We find that the index of proportional representation of species (alpha index) for the forest growing from clear-cuts increases significantly under climate change, but decreases under climate change plus logging. The index of changing representation of species (beta(c) index) increases significantly under climate change and climate change plus logging. When any one species is removed alpha diversity of the forest growing from clear-cuts changes significantly under climate change, but beta(c) index remains almost the same. When all individuals of shade tolerant species, shade intolerant species, or medium type species are removed, respectively, alpha diversity decreases, but beta(c) diversity changes in more complicated ways. The implications of these results for preserving tree diversity in this type of forest are also discussed. PMID- 12591255 TI - Recent findings on natural products with erectile-dysfunction activity. AB - Viagra has had a profound influence on the search for natural products with erectile-dysfunction activity. To date the "natural" equivalent is not in existence but several pure compounds from nature, e.g., Yohimbine, Citrulline, two pyrano-isoflavones, berberine, forskolin and others, have either been re examined or are new potential candidates. Intense activity exists in the area of testing semi-purified plant extracts for erectile dysfunction activity. PMID- 12591256 TI - The chemical characteristic and distribution of brassinosteroids in plants. AB - Brassinosteroids represent a class of plant hormones with high-growth promoting activity. They are found at low levels in pollen, anthers, seeds, leaves, stems, roots, flowers, grain, and young vegetative tissues throughout the plant kingdom. Brassinosteroids are a family of about 60 phytosteroids. The article gives a comprehensive survey on the hitherto known brassinosteroids isolated from plants. The chemical characteristic of brassinosteroids is also presented. PMID- 12591257 TI - Biochemical evidence for a calcium-dependent protein kinase from Pharbitis nil and its involvement in photoperiodic flower induction. AB - A soluble Ca(2+)-dependent protein kinase (CDPK) was isolated from seedlings of the short-day plant Pharbitis nil and purified to homogeneity. Activity of Pharbitis nil CDPK (PnCDPK) was strictly dependent on the presence of Ca(2+) (K(0,5)=4,9 microM). The enzyme was autophosphorylated on serine and threonine residues and phosphorylated a wide diversity of substrates only on serine residues. Histone III-S and syntide-2 were the best phosphate acceptors (K(m) for histone III-S=0,178 mg ml(-1)). Polyclonal antibodies directed to a regulatory region of the soybean CDPK recognized 54 and 62 kDa polypeptides from Pharbitis nil. However, only 54 kDa protein was able to catalyse autophosphorylation and phosphorylation of substrates in a Ca(2+)-dependent manner. CDPK autophosphorylation was high in 5-day-old Pharbitis nil seedlings grown under non inductive continuous white light and was reduced to one-half of its original when plants were grown in the long inductive night. Also, the pattern of proteins phosphorylation has changed. After 16-h-long inductive night phosphorylation of endogenous target (specific band of 82 kDa) increased in the presence of calcium ions. It may suggest that Ca(2+)-dependent protein kinase is involved in this process and it is dependent on light/dark conditions. PMID- 12591258 TI - Procerain, a stable cysteine protease from the latex of Calotropis procera. AB - A protease was purified to homogeneity from the latex of medicinal plant Calotropis procera (Family-Asclepiadaceae). The molecular mass and isoelectric point of the enzyme are 28.8 kDa and 9.32, respectively. Hydrolysis of azoalbumin by the enzyme was optimal in the range of pH 7.0-9.0 and temperature 55-60 degree C. The enzyme hydrolyses denatured natural substrates like casein, azoalbumin, and azocasein with high specific activity. Proteolytic and amidolytic activities of the enzyme were activated by thiol protease activators and inhibited by thiol protease inhibitors, indicating the enzyme to be a cysteine protease. The enzyme named as procerain, cleaves N-succinyl-Ala-Ala-Ala-p-nitroanilide but not -Ala Ala-p-nitroanilide, -Ala p-nitroanilide and N-d-Benzoyl--Arg-p-nitroanilide and appears to be peptide length dependent. The extinction coefficient (epsilon 1% 280 nm) of the enzyme was 24.9 and it had no detectable carbohydrate moiety. Procerain contains eight tryptophan, 20 tyrosine and seven cysteine residues forming three disulfide bridges, and the remaining one being free. Procerain retains full activity over a broad range of pH 3.0-12.0 and temperatures up to 70 degree C, besides being stable at very high concentrations of chemical denaturants and organic solvents. Polyclonal antibodies against procerain do not cross-react with other related proteases. Procerain unlike most of the plant cysteine proteases has blocked N-terminal residue. PMID- 12591259 TI - Pn-AMPs, the hevein-like proteins from Pharbitis nil confers disease resistance against phytopathogenic fungi in tomato, Lycopersicum esculentum. AB - The antifungal activity of hevein-like proteins has been associated with their chitin-binding activities. Pn-AMP1 and Pn-AMP2, two hevein homologues from Pharbitis nil, show in vitro antifungal activities against both chitin and non chitin containing fungi. Purified Pn-AMPs retained antifungal activities only under non-reducing conditions. When Pn-AMP2 cDNA was constitutively expressed in tomato (Lycopersicon esculentum) plants under the control of CaMV35S promoter, the transgenic plants showed enhanced resistance against both the non-chitinous fungus Phytophthora capsici, and the chitin-containing fungus Fusarium oxysporum. Thus, the chitin component in the fungal cell wall is not an absolute requirement for Pn-AMP's antifungal activities. These results when considered together suggest that Pn-AMPs have the potential for developing transgenic plants resistant to a wide range of phytopathogenic fungi. PMID- 12591260 TI - A cDNA clone for 3-carene synthase from Salvia stenophylla. AB - The essential oil of Salvia stenophylla contains (+)-3-carene as the principal monoterpene component. Using an enriched cDNA library prepared from mRNA isolated from S. stenophylla peltate glandular trichomes, and a homology-based cloning strategy, a full-length cDNA was isolated that encoded a preprotein of 69.7 kDa which resembled a monoterpene synthase in sequence. Heterologous expression of the gene in Escherichia coli provided a soluble recombinant enzyme capable of catalyzing the divalent metal ion-dependent conversion of geranyl diphosphate to (+)-3-carene and to lesser amounts of limonene, myrcene, 4-carene and beta phellandrene. This multiple-product synthase is responsible for the production of all of the essential oil monoterpenes of S. stenophylla. PMID- 12591261 TI - Neuroactive and other free amino acids in seed and young plants of Panax ginseng. AB - The seeds and one to three years old plants of Asian ginseng (Panax ginseng C.A. Meyer) were analyzed for their free amino acid contents. The neuro-excitatory beta-ODAP (beta-N-oxalyl-L-alpha,beta-diaminopropionic acid), suggested to be the cause of the crippling neurolathyrism, was the major component in the seed extract (70% of the total free amino acids detected) and showed the highest concentration (0.43% by wt) compared to that in the different parts of young plants. beta-ODAP concentration was higher in the shoots as compared to roots and declined in older plants. The amount of beta-ODAP in the roots may be considered as an indirect measure of age and quality. Another neuro-active non-protein amino acid, GABA (gamma-aminobutyric acid), increased dramatically after germination and reached highest concentration in different parts of 3 year-old plants. Glutamine and arginine were the two major free proteinogenic amino acids in the ginseng plants and together they constituted over 50% of all the free amino acids detected in the root. PMID- 12591262 TI - Efficient production and capture of 8-prenylnaringenin and leachianone G biosynthetic intermediates of sophoraflavanone G--by the addition of cork tissue to cell suspension cultures of Sophora flavescens. AB - It has previously been demonstrated that the addition of cork tissue to cell suspension cultures of Sophora flavescens stimulates the production of sophoraflavanone G, most of which has been recovered from the added cork tissue. In the present study, it was found that two precursors of sophoraflavanone G, 8 prenylnaringenin (sophoraflavanone B) and leachianone G, both of which have never been detected either in cultured cells or in the original plants, also accumulated in the added cork tissue. Thirteen minor flavonoids including three prenylated flavonoids, in addition to 8-prenylnaringenin and leachianone G, were isolated from the cork tissue co-incubated with S. flavescens cells. The new compounds flavescenones A, B and C, were determined to be (3R)-5, 7, 2' trihydroxy-6-gamma, gamma-dimethylallyl-4', 5'-methylenedioxyisoflavanone; 5, 7, 2'-trihydroxy-6-gamma, gamma-dimethylallyl-4', 5'-methylenedioxyisoflavone and 2 [2',4'-dihydroxy-3'-(gamma-hydroxymethyl-gamma-methylallyl)phenyl]-5,6 methylenedioxybenzofuran, respectively, by means of spectroscopic analyses that included 2D-NMR techniques. PMID- 12591263 TI - Microbial transformation of zaluzanin-D. AB - Microbial transformation of zaluzanin-D using different fungi gave 11,13 dihydrozaluzanin-C, zaluzanin-C, 4,16,11,13 - tetrahydro zaluzanin-C, estafiatone, dihydroestafiatol and dihydroestafiatone. PMID- 12591264 TI - Structure, chemistry, and biological activity of pseudophomins A and B, new cyclic lipodepsipeptides isolated from the biocontrol bacterium Pseudomonas fluorescens. AB - Pseudophomins A and B are cyclic lipodepsipeptides isolated from Pseudomonas fluorescens strain BRG100, a bacterium with potential application for biocontrol of plant pathogens and weeds. Their chemical structures were established by a combination of spectroscopic data, X-ray crystallography, and selective chemical degradation. This unique chemical degradation allowed the unambiguous determination of the absolute configuration of the amino acid residue Leu-1, due to gamma-lactam formation followed by selective cleavage of the adjacent N(8) C(7) bond. To the best of our knowledge this is the first application of gamma lactam formation to the determination of absolute configuration of an adjacent amino acid. Pseudophomin B showed higher antifungal activity against the phytopathogens Phoma lingam/Leptosphaeria maculans and Sclerotinia sclerotiorum than pseudophomin A, and is likely to be the main component responsible for the antifungal activity of EtOAc extracts of strain BRG100. By contrast, pseudophomin A showed stronger inhibition of green foxtail (Setaria viridis) root germination than pseudophomin B. PMID- 12591265 TI - (+)-Strigol, a witchweed seed germination stimulant, from Menispermum dauricum root culture. AB - (+)-Strigol was isolated from Menispermum dauricum root culture filtrate. Its identity was confirmed by HPLC, 1H NMR, UV and MS, and on the basis of its CD spectrum. This is the first report on isolation of strigolactone from aseptic plant culture. PMID- 12591266 TI - Meliternatin: a feeding deterrent and larvicidal polyoxygenated flavone from Melicope subunifoliolata. AB - We screened more than 60 Malaysian plants against two species of insects and found that Melicope subunifoliolata (Stapf) T.G. Hartley (Rutaceae) showed strong feeding deterrent activity against Sitophilus zeamais Motsch. (Curculionidae) and very good larvicidal activity against Aedes aegypti L. (Diptera). One anti-insect compound, meliternatin (3,5-dimethoxy-3',4',6,7-bismethylendioxyflavone) (6) and six other minor polyoxygenated flavones were isolated from M. subunifoliolata. PMID- 12591267 TI - Taxonomic significance of flavonoid variation in temperate species of Nothofagus. AB - Forty-two flavonoids and a stilbene (pinosylvin) were identified in the leaf exudates of 11 temperate species of Nothofagus from South America, Australia and New Zealand. The flavonoid profiles demonstrate significant taxonomic value at the subgeneric level. Most species of subgenus Fuscospora are characterized by the presence of pinosylvin, galangin and galangin methyl ethers. Kaempferol-type flavonols are abundant in subgenus Lophozonia while these flavonols are largely absent from species of subgenus Fuscospora. The flavonoid patterns are largely in agreement with a recent subgeneric classification of Nothofagus. PMID- 12591268 TI - A plant growth retardant related to chlamydocin and its proposed mechanism of action. AB - A comparison of the plant growth retardant activity of the chlamydocin analogues, compound 1, six derivatives from 1 and 2, and two synthetic analogues revealed that there are two types of retardant in chlamydocin analogues. One, for example in compound 1, requires an oxygen atom at C-8 of the 2-aminodecanoic acid moiety to show retardant activity. The other, for example in compound 8, requires no oxygen atom at C-8 but requires a specific alkyl group chain length for activity. To determine the differences in mode of action of both types of retardant, rice seedlings were separately treated with compounds 1 and 8, and after appearance of dwarfism, their endogenous ABA and GA(1) levels were determined and compared to those of the control. Treatment with 1 (10 nmol/plant) increased ABA levels 4 times higher than that of the control and decreased GA(1) levels to 20% of that of the control. Treatment with 8 (30 nmol/plant) did not affect the ABA level but decreased GA(1) content to 5% of that of the control. PMID- 12591269 TI - Isolation and identification of an allelopathic substance in Pisum sativum. AB - The residue of peas (Pisum sativum L.) has allelopathic activity and the putative compound causing this inhibitory effect was isolated from a methanol extract of pea shoots. Chemical structure of this compound was determined by high-resolution MS, IR and 1H NMR spectral data as pisatin. Pisatin inhibited growth of cress (Lepidium sativum L.) and lettuce (Lactuca sativa L.) seedlings at concentrations greater than 10 and 30 microM, respectively. The doses required for 50% growth inhibition of roots and hypocotyls of cress were 61 and 91 microM, respectively, and those of lettuce were 78 and 115 microM, respectively. The concentration of pisatin in the pea shoots was 32.7 nmol x g(-1) fresh weight. The effectiveness of pisatin on growth inhibition in cress and lettuce, and its occurrence in pea shoots suggest that it may contribute to the growth inhibitory effect of pea residue, and may play an important role in pea allelopathy. PMID- 12591270 TI - Effects of seven pure flavonoids from mosses on germination and growth of Tortula muralis HEDW (Bryophyta) and Raphanus sativus L (Magnoliophyta). AB - Dried mosses (five moss species) were progressively extracted and subjected to a four-step Craig distribution. Seven pure flavonoids were isolated and identified. The flavonoids were the flavones apigenin, apigenin-7-O-triglycoside, lucenin-2, luteolin-7-O-neohesperidoside, saponarine and vitexin; and the biflavonoid bartramiaflavone and they were submitted to biological tests. The tests were performed in vitro on spore germination and protonemal growth of the moss Tortula muralis and on seed germination and root growth of Raphanus sativus. Flavonoids caused a decrease in the percentage of spore germination, protonemal development and root growth. In addition they caused morphological alterations, such as forked tips, swollen apices, rounded cells and early formation of brood cells in the protonemata. Data were discussed in relation to the presence of allelochemicals in mosses. PMID- 12591271 TI - Homoisoflavonoids from Ophiopogon japonicus Ker-Gawler. AB - From the ethyl acetate extract of the tuberous roots of Ophiopogon japonicus (Liliaceae) eight known and five new homoisoflavonoidal compounds were isolated. The new compounds are 5,7-dihydroxy-8-methoxy-6-methyl-3-(2'-hydroxy-4' methoxybenzyl)chroman-4-one (1), 7-hydroxy-5,8-dimethoxy-6-methyl-3-(2'-hydroxy 4'-methoxybenzyl)chroman-4-one (2), 5,7-dihydroxy-6,8-dimethyl-3-(4'-hydroxy-3' methoxybenzyl)chroman-4-one (3), 2,5,7-trihydroxy-6,8-dimethyl-3-(3',4' methylenedioxybenzyl)chroman-4-one (4) and 2,5,7-trihydroxy-6,8-dimethyl-3-(4' methoxybenzyl)chroman-4-one (5). Their structures have been elucidated by mass and NMR spectroscopy. Compounds 4 and 5 are the first isolated homoisoflavonoids with a hemiacetal function at position 2. PMID- 12591272 TI - MALDI-TOF mass spectrometry and PSD fragmentation as means for the analysis of condensed tannins in plant leaves and needles. AB - MALDI-TOF mass spectrometry and 13C NMR spectroscopy were applied to unveil typical characteristics of condensed tannins of leaves and needles from willow (Salix alba), spruce (Picea abies) and beech (Fagus sylvatica) of three tree species that are ubiquitous in German forests and landscapes. For further evaluation, lime (Tilia cordata) was included. The 13C NMR spectroscopy confirmed the purity of the condensed tannin fractions and the efficiency of the procedure used for their extraction. While signals representative for procyanidin units are observable in all liquid-state 13C NMR spectra, resonance lines of prodelphinidin were only detected in those obtained from the condensed tannins of spruce needles and beech leaves. Typical signals in the chemical shift region between 70 and 90 ppm demonstrated the presence of stereoisomers (catechin/epicatechin; gallocatechin/ epigallocatechin). The MALDI-TOF mass spectra of the condensed tannins show signals of polymers of up to undecamers. Supporting the observations from the NMR spectroscopy, the mass spectra of the willow and lime leaf condensed tannins were identified as polymers with mainly procyanidin units, while the polymers of the spruce needle and beech leaves exhibit varying procyanidin/prodelphinidin ratios. Post source decay (PSD) fragmentation lead to a sequential loss of monomers and allowed a detailed characterization and sequencing of individual chains. In the case of the condensed tannins of lime this technique clearly excludes a pelargonidin terminal unit followed by a prodelphinidin unit, which would result in the same molecular masses as a polymer solely built up of procyanidin units. PMID- 12591274 TI - High level of transgene expression in primary chronic lymphocytic leukemia cells using helper-virus-free recombinant Epstein-Barr virus vectors. AB - OBJECTIVE: Epstein-Barr virus (EBV)-based vectors have favorable features for gene transfer, including a high transduction efficiency especially for B cells, large packaging capacity up to 150 kb pairs, and ability to infect postmitotic cells. Recombinant EBV was explored for transduction of primary human B-cell chronic lymphocytic leukemia (CLL) cells. MATERIAL AND METHODS: EBV vectors deleted for all oncogenic sequences and encoding terminal repeats (TR) essential for encapsidation, the lytic origin of replication (oriLyt) for DNA amplification, and the enhanced green fluorescent protein (EGFP) were packaged using an optimized, helper-virus-free method. Infectious EBV virions encoding EGFP (EBV/EGFP) with an infectious titer up to 2 x 10(6) per milliliter were generated. Primary leukemic cells from 14 patients with CLL were successfully transduced with EBV/EGFP at a very low multiplicity of infection (< 1). RESULTS: Transgene expression was detected in up to 85% of cells 48 hours after infection. Transduction was specifically mediated by EBV vectors because gene transfer was inhibited by an antibody (72A1) directed against the viral envelope glycoprotein gp350/220. Furthermore, transduction of CLL cells with packaged EBV vectors coding for EGFP but deleted for TR sequences (TR-) did not result in EGFP expression compared to TR+ vector constructs (p = 0.009). CONCLUSION: Helper virus-free EBV-based gene transfer vectors hold promise for development of genetic therapies for CLL patients. PMID- 12591275 TI - Granulocyte colony-stimulating factor induces the release in the bone marrow of proteases that cleave c-KIT receptor (CD117) from the surface of hematopoietic progenitor cells. AB - OBJECTIVE: Administration of granulocyte colony-stimulating factor (G-CSF) results in the mobilization of hematopoietic progenitor and stem cells from the bone marrow into the peripheral blood. Although the mechanisms leading to the mobilization of primitive hematopoietic cells is not fully understood, it has been noted that the yield of mobilization in humans is correlated to the down regulation of c-KIT/CD117 expression on mobilized cells. We sought to determine the mechanisms responsible for the reduced expression of c-KIT on mobilized hematopoietic progenitor cells. MATERIALS AND METHODS: Mice were mobilized with G CSF and primitive hematopoietic cells were collected from bone marrow and blood to analyze c-KIT expression. Using cell lines expressing mouse and human c-KIT and a recombinant protein comprising the entire extracellular domain of human c KIT, we analyzed by flow cytometry and immunoblotting the proteolytic cleavage of c-KIT by proteases released in bone marrow extracellular fluids extracted from mobilized mice. RESULTS: Administration of G-CSF into mice results in the reduction of c-KIT expression on primitive hematopoietic cells in bone marrow and peripheral blood. Bone marrow extracellular fluids isolated from G-CSF-mobilized mice contain serine proteases that cleave c-KIT into discrete fragments. Proteases capable of cleaving c-KIT include neutrophil elastase, cathepsin G, proteinase-3 and matrix metalloproteinase-9. CONCLUSIONS: In addition to transcriptional controls, exocytosis, and ligand-induced internalization, the direct proteolytic cleavage of c-KIT by neutrophil and macrophage proteases represents a novel pathway to regulate the levels of c-KIT expression at the surface of hematopoietic cells and may be responsible in part for the down regulation of c-KIT expression on mobilized hematopoietic progenitors in vivo. PMID- 12591276 TI - Fluorescence in situ hybridization analysis of the PRV-1 gene in polycythemia vera: implications for its role in diagnosis and pathogenesis. AB - OBJECTIVE: In polycythemia vera (PV) there is no specific clonal marker because the molecular lesion responsible for PV is unknown. The recent demonstration that the PRV-1 gene is overexpressed in granulocytes from patients with PV provided the rationale for the current study to investigate whether PRV-1 is structurally rearranged, thus explaining its aberrant expression. MATERIALS AND METHODS: Fluorescence in situ hybridization was used to determine chromosomal localization of PRV-1 and to study whether the PRV1 gene is rearranged in 26 patients with PV. RESULTS: PRV-1 was localized to chromosome 19, band region q13.12-2. Structural rearrangements of PRV-1 were evaluated in bone marrow cells from 26 patients with PV: 14 with a normal karyotype and 12 with an abnormal karyotype. None of 150 metaphase cells or more than 10,000 interphase cells demonstrated PRV-1 gene deletion, amplification, or separation of the probe signal, which would indicate a PRV-1 rearrangement. CONCLUSION: These findings are consistent with a lack of structural rearrangement of PRV-1 in patients with PV. Thus, overexpression of PRV-1 in granulocytes from patients with PV is related to mechanisms that do not involve structural genetic changes. PMID- 12591277 TI - Inverse relationship between myeloid maturation and leukotriene C4 synthase expression in normal and leukemic myelopoiesis-consistent overexpression of the enzyme in myeloid cells from patients with chronic myeloid leukemia. AB - OBJECTIVE: Leukotriene (LT) C(4) synthase (LTC(4)S) is the key enzyme in the biosynthesis of LTC(4), which has been reported to stimulate the growth of human myeloid progenitor cells and is specifically overproduced in chronic myeloid leukemia (CML). The aim of this study was to clarify the expression of LTC(4)S during normal and leukemic myelopoiesis and to investigate the correlation between abnormal LTC(4)S expression in CML myeloid cells and the activity of the disease-specific tyrosine kinase p210 BCR-ABL. MATERIALS AND METHODS: Immature and mature myeloid cell subpopulations were isolated with magnetic cell sorting from healthy volunteer bone marrow (n = 11) and CML patient peripheral blood (n = 8), respectively. The cells were subjected to analysis of LTC(4)S protein expression and activity. Expression of LTC(4)S was investigated in CD16(+) neutrophils from CML patients before and after 1 month of medication with imatinib mesylate (STI571), which is a specific inhibitor of p210 BCR-ABL. RESULTS: Among normal cells, the highest enzyme activity was observed in the most immature, CD34(+) progenitor cell-enriched and CD15(+) myelocyte-enriched fractions. Subsequently, LTC(4)S activity decreased with increasing maturity, with only negligible amounts of LTC(4) produced in CD16(+) neutrophils. LTC(4)S was expressed at the protein level in the immature myeloid cell fractions but not in CD16(+) cells. In CML cells, LTC(4)S activity and expression were consistently elevated. Thus, the CML CD34(+) and CD15(+) cell fractions, as well as the CD11b(+) myelocyte/metamyelocyte-enriched fractions, produced 6 to 10 times as much LTC(4) as the corresponding normal cells. Again, enzyme expression was highest in the most immature cells, although evident LTC(4)S expression and activity remained in CML CD16(+) neutrophils. Interestingly, treatment of five CML patients with imatinib mesylate down-regulated the abnormal neutrophil LTC(4)S expression and activity. CONCLUSIONS: Expression of LTC(4)S in immature myelopoid cells is in line with a role for this enzyme in myelopoiesis. In addition, consistent overexpression of LTC(4)S in CML and the correlation to p210 BCR-ABL activity suggests that LTC(4)S may be involved in leukemic pathogenesis. PMID- 12591278 TI - Abnormal splicing of SHP-1 protein tyrosine phosphatase in human T cells. Implications for lymphomagenesis. AB - OBJECTIVE: SHP-1 protein tyrosine phosphatase has been implicated in suppressing B-lymphocyte and myeloid cell malignancies; however, there are little data on this role of SHP-1 in T-lymphocyte malignancies. We examined malignant human T cells to identify any abnormalities of SHP-1 that would support a role for this molecule in suppressing T lymphomagenesis. MATERIALS AND METHODS: Human T lymphocyte cell lines and primary blood cells were used to examine the expression of SHP-1 mRNA and protein. Reverse transcriptase polymerase chain reaction was used to amplify particular portions of the SHP-1 mRNA for cloning and sequencing. Gene transfer was used to examine the effects of SHP-1 on cell growth and morphology. Glutathione S-transferase (GST) fusion proteins were generated and used to determine SHP-1-associated proteins. RESULTS: Leukemia- and lymphoma derived cell lines were identified that did not express SHP-1 protein. Examination of the mRNA from these and other T-cell lines, and from normal peripheral blood mononuclear cells (PBMCs), revealed three distinct transcripts by restriction enzymes, reverse transcriptase polymerase chain reaction, and Southern blot analysis. In addition to the expected wild-type transcript, two novel transcripts were identified. One was a deletion transcript found only in Jurkat leukemia-derived cells, predicted to encode for a 7-kDa protein containing most of the amino-terminal SH2 domain. The second contained an 88-nucleotide insert that is the unspliced second intron resulting in a frame shift and the formation of a noncoding transcript. This mRNA was found in all cells examined but was the only transcript detected in the cell lines lacking SHP-1 protein. Expressing wild-type SHP-1 in these cell lines resulted in a change in the morphology of the cells with a concomitant decrease in their growth. GST fusion constructs showed the 7-kDa variant able to associate with an identical array of proteins as wild-type SHP-1, suggesting that it could compete with the wild-type SHP-1 for substrates. This protein was detectable in the cell line expressing its corresponding mRNA and was able to induce significant changes in cell morphology when transfected into a cell line expressing wild-type SHP-1; however, it did not induce any changes in cell growth. CONCLUSIONS: These data are the first to show the existence of multiple transcripts of SHP-1 in human transformed T lymphocytes and normal PBMCs and supports previous work showing that alternate forms of SHP-1 mRNA are a common finding in other cells. We also show the lack of splicing out of an intron as a novel mechanism of regulation of SHP-1 protein expression in both normal and transformed T cells. Moreover, we provide the first evidence showing a protein product detectable in cells that is translated from an alternatively spliced form of SHP-1 mRNA, a variant truncated SHP-1 protein having potential biologic relevance. This report provides evidence supporting the concept that SHP-1 can negatively regulate growth of malignant human T cells and that lack of SHP-1 protein or function may be associated with lymphomagenesis. PMID- 12591280 TI - X-linked thrombocytopenia caused by a mutation in the Wiskott-Aldrich syndrome (WAS) gene that disrupts interaction with the WAS protein (WASP)-interacting protein (WIP). AB - OBJECTIVE: We studied two adult brothers with severe congenital thrombocytopenia in order to determine the genetic etiology of their inherited disorder. Despite the absence of eczema or immunodeficiency, a mutation of the Wiskott-Aldrich syndrome (WAS) gene was suspected because of the presence of microthrombocytes. MATERIALS AND METHODS: Peripheral blood was obtained for characterization of hematopoietic cells and megakaryocyte progenitors. The coding region of the WAS gene was fully sequenced, and expression of the Wiskott-Aldrich syndrome protein, WASP, was evaluated by immunoblotting. The ability of WASP to physically associate with the WASP-interacting protein, WIP, was tested by yeast and mammalian two-hybrid techniques. RESULTS: In addition to thrombocytopenia, our investigation revealed an increased frequency of peripheral megakaryocyte progenitors (CFU-Mk) and incomplete cytoplasmic maturation by electron microscopy. Sequencing the WAS gene revealed a single base mutation, resulting in substitution of proline for arginine 138 (i.e., Arg138Pro). Immunoblotting demonstrated reduced expression of the mutant WAS protein, and we showed that the Arg138Pro mutation significantly, but incompletely, disrupts WASP-WIP interaction. CONCLUSIONS: In this pedigree, X-linked thrombocytopenia is caused by a rare mutation in the fourth exon of the WAS gene. WASP levels are reduced in lymphocyte cell lines derived from the affected individuals. Furthermore, the mutation significantly but incompletely disrupts WASP-WIP interaction, whereas substitution of alanine or glutamic acid residues at the same position does not. This raises the possibility that protein-protein interaction and WASP stability are related properties. PMID- 12591279 TI - Antitumor activity of the angiogenesis inhibitor TNP-470 on murine lymphoma/leukemia cells in vivo and in vitro. AB - OBJECTIVE: The aim of this study was to evaluate the effects of an angiogenesis inhibitor in a non-immunocompromised setting in which transplanted tumor cells home and expand in a manner mimicking the original tumor in the donor. We used a novel animal model for T-cell lymphoma/leukemia (TLL) to test the antitumor effect of TNP-470, a well-established angiogenesis inhibitor. MATERIALS AND METHODS: Cells from spontaneously arisen TLL tumors were transferred to syngenic recipients. The mice were treated with TNP-470 (30 mg/kg) or vehicle every other day for 2 weeks. Mice were sacrificed on day 15 after transfer, and body and organ weights were measured. Cell cycle and morphologic analysis was performed on cells and/or sections from selected organs. The cytotoxic effect of TNP-470 was assayed in vitro using the TLL-M and HL-60 cell lines. RESULTS: TNP-470 treatment significantly reduced total tumor load and tumor mass in specific organs infiltrated with lymphoma/leukemia. This was associated with an increased apoptosis in these organs. We also observed side effects of TNP-470 not previously reported, such as diminished extramedullary erythropoiesis and disrupted liver morphology. In vitro TLL-M cells were resistant to cytotoxic effects of moderate doses of TNP-470. CONCLUSIONS: TNP-470 treatment has a beneficial effect on tumor load in the TLL transfer model, most likely caused by the antiangiogenic effect of TNP-470. This is supported by the observation of increased apoptosis in infiltrated organs. The TLL transfer model is well suited for further studies of combinations with TNP-470 or other angiogenesis inhibitors and cytotoxic drugs. PMID- 12591281 TI - Enhanced green fluorescent protein targeted to the Sca-1 (Ly-6A) locus in transgenic mice results in efficient marking of hematopoietic stem cells in vivo. AB - OBJECTIVE: Hematopoietic stem cells are important clinically, both as targets of disease and as reagents for cellular therapy. Studies in hematopoietic stem cell biology have been hampered by difficulties in purifying and manipulating these cells. To facilitate these studies, we sought to develop a system for targeting genes of interest to the hematopoietic stem cell compartment in transgenic mice. MATERIALS AND METHODS: We used Sca-1, a glycosyl phosphatidylinositol-anchored protein expressed on the surface of all hematopoietic stem cells in commonly used inbred mouse strains. We created a mutant Sca-1 allele in which the enhanced green fluorescent protein (EGFP) cDNA is integrated into the Sca-1 locus by homologous recombination in embryonic stem cells. RESULTS: EGFP protein is detectable in all hematopoietic tissues of mice heterozygous for the mutant Sca-1 allele. Growth and development of these mice are normal. No adverse effects of long-term, high-level EGFP expression were noted. Sca-1 positive cells coexpress EGFP in all tissues and lineages examined, as predicted by the targeting strategy. Sca-1 and EGFP expression are coordinately up-regulated in splenocytes from mutant mice. The Lin(-)EGFP(+) bone marrow population contains all progenitor activity in Sca-1(+)(/EGFP) mice. The Lin(-)EGFP(+) bone marrow cells are equivalent to Lin(-)Sca-1(+) cells in long-term repopulation and serial transplantation assays. CONCLUSION: The hematopoietic stem cell compartment appears to be targeted in Sca-1(+)(/EGFP) mutant mice. This system should be useful for studying the normal biology of hematopoietic stem cells and for targeting other genes to this cellular compartment. PMID- 12591282 TI - Dog leukocyte antigen-haploidentical stem cell allografts after anti-CD44 therapy and reduced-intensity conditioning in a preclinical canine model. AB - OBJECTIVE: We previously described a nonmyeloablative hematopoietic stem cell transplantation regimen in dog leukocyte antigen (DLA)-identical littermate recipients consisting of low-dose total body irradiation (TBI) before and mycophenolate mofetil (MMF)/cyclosporine (CSP) given after transplant to control both graft-vs-host and residual host-vs-graft reactions. In this study, we sought to develop a reduced-intensity regimen to achieve engraftment across major histocompatibility complex barriers in DLA-haploidentical littermate recipients. MATERIALS AND METHODS: We tested a regimen of 450-cGy TBI with or without postgrafting MMF/CSP for 28 and 35 days, respectively, and with the administration of monoclonal antibody (mAb) S5 (anti-CD44), at a dose of 0.2 mg/kg/day from days -7 through -2, prior to receiving TBI. RESULTS: One of six dogs conditioned with 450-cGy TBI alone achieved engraftment of granulocyte colony-stimulating factor-mobilized peripheral blood stem cells. Three of six dogs achieved sustained donor cell engraftment using 450-cGy TBI and posttransplantation MMF/CSP. None of three dogs given mAb S5 followed by 450-cGy TBI showed signs of donor cell engraftment. However, when S5 mAb pretreatment was added to 450-cGy TBI and postgrafting MMF/CSP, 10 of 12 dogs achieved sustained engraftment (p = 0.008 or 0.007 vs 450-cGy alone or to S5 + 450-cGy TBI without MMF/CSP, respectively), with only three dogs developing severe graft-vs-host disease on this short regimen of immunosuppression. CONCLUSION: These results show that engraftment across a DLA haplotype-mismatched barrier can be achieved after reduced-intensity conditioning when mAb S5 directed at CD44 is added to this regimen. PMID- 12591283 TI - Mixed chimerism following in utero hematopoietic stem cell transplantation in murine models of hemoglobinopathy. AB - OBJECTIVE: Mixed hematopoietic chimerism after bone marrow transplantation can provide effective treatment for beta-thalassemia because of the selective advantage that exists for donor erythropoiesis. In utero hematopoietic stem cell transplantation (IUHSCTx) can achieve mixed hematopoietic chimerism, particularly when a selective advantage exists for donor cells. To investigate the biology of IUHSCTx in hemoglobinopathies, we performed fully allogeneic IUHSCTx in murine models of beta-thalassemia (Thal) and sickle cell disease (SCD). MATERIALS AND METHODS: We serially assessed and compared levels of mononuclear cell (MNC) and erythroid chimerism after IUHSCTx of either adult bone marrow (BM)- or fetal liver (FL)-derived allogeneic donor cells in the two hemoglobinopathy models, which differ significantly in their degree of anemia (Thal>>SCD) and red cell half-life (Thal< veins approximately gray matter > white matter. Mean delays displayed the same ranking across tissue types (p < 0.001), except that veins > gray matter. However, measures for all tissue types were distributed across an overlapping range. A logistic regression model correctly discriminated 72% of the veins from gray matter in the absence of independent information of macroscopic vessels (ROC = 0.72). While tissue classification was not perfect for this subject, weighting the T contrast by the predicted probabilities materially reduced the venous component to the activation map. PMID- 12591566 TI - Functional magnetic resonance imaging of tonic pain and vasopressor effects in rats. AB - In functional magnetic resonance imaging (fMRI) studies, an elevation in blood pressure (BP) in individuals with a poor autoregulatory response may increase cerebral blood flow, potentially enhancing the blood oxygenation level dependent response. To investigate the role of BP changes, the cerebral activation to either tonic pain or the infusion of the vasopressor norepinephrine was correlated with the accompanying BP changes in alpha-chloralose anesthetized rats. Immediately after formalin (2%) injection into the forepaw, fMRI detected an activation that was correlated with the BP increase and additional activations that were independent of blood pressure changes 5-40 minutes later. The activation detected with the administration of the vasopressor norepinephrine, which does not cross the blood-brain barrier was correlated to both the amount and rate of increase in BP. The response ranged from being sparse, localized within cortex or widespread during modest, moderate or severe elevations in BP, respectively. The cerebral circulatory effects of hypertension should be considered as contributing to changes in cerebral blood oxygenation in fMRI studies involving increases in BP. PMID- 12591567 TI - Brain MRI lesion load quantification in multiple sclerosis: a comparison between automated multispectral and semi-automated thresholding computer-assisted techniques. AB - Brain magnetic resonance imaging (MRI) lesion volume measurement is an advantageous tool for assessing disease burden in multiple sclerosis (MS). We have evaluated two computer-assisted techniques: MSA multispectral automatic technique that is based on bayesian classification of brain tissue and NIH image analysis technique that is based on local (lesion by lesion) thresholding, to establish reliability and repeatability values for each technique. Brain MRIs were obtained for 30 clinically definite relapsing-remitting MS patients using a 2.0 Tesla MR scanner with contiguous, 3 mm thick axial, T1, T2 and PD weighted modalities. Digital (Dicom 3) images were analyzed independently by three observers; each analyzed the images twice, using the two different techniques (Total 360 analyses). Accuracy of lesion load measurements using phantom images of known volumes showed significantly better results for the MSA multispectral technique (p < 0.001). The mean intra-and inter-observer variances were, respectively, 0.04 +/- 0.4 (range 0.04-0.13), and 0.09 +/- 0.6 (range 0.01-0.26) for the multispectral MSA analysis technique, 0.24 +/- 2.27 (range 0.23-0.72) and 0.33 +/- 3.8 (range 0.47-1.36) for the NIH threshold technique. These data show that the MSA multispectral technique is significantly more accurate in lesion volume measurements, with better results of within and between observers' assessments, and the lesion load measurements are not influenced by increased disease burden. Measurements by the MSA multispectral technique were also faster and decreased analysis time by 43%. The MSA multispectral technique is a promising tool for evaluating MS patients. Non-biased recognition and delineation algorithms enable high accuracy, low intra-and inter-observer variances and fast assessment of MS related lesion load. PMID- 12591568 TI - Precise estimate of fundamental in-vivo MT parameters in human brain in clinically feasible times. AB - A methodology is presented for extracting precise quantitative MT parameters using a magnetisation-prepared spoiled gradient echo sequence. This method, based on a new mathematical model, provides relaxation parameters for human brain in vitro and in-vivo. The in-vivo parameters have been obtained from three different regions of normal white matter: occipital white matter, frontal white matter and centrum semiovale; two regions of normal grey matter: cerebral cortex and cerebellum, and from five regions with MS lesions. All this has been achieved using MT images collected within a timeframe that is clinically feasible. We hope that this new technique will shed light on the properties and dynamics of water compartments within the brain. PMID- 12591569 TI - T2 relaxation time histograms in multiple sclerosis. AB - An accurate measurement of the transverse relaxation time T(2), and the histogram of T(2) in the brain parenchyma can be accomplished in vivo using a multi-echo magnetic resonance imaging sequence. An estimate of the error in the T(2) measurement is derived using copper sulfate doped water phantoms. Correction factors are calculated and applied to the signal intensity of each voxel prior to the in vivo T(2) evaluation. These corrected T(2) are in good agreement with the theoretical values calculated from copper sulfate concentrations. This technique is then applied to calculate T(2) histograms of the brain. The population studied was composed of normal volunteers and multiple sclerosis patients. The corrected T(2) histogram method discriminates the normal control population from the MS population, and also discriminates between relapsing-remitting patients and primary progressive or secondary progressive patients. Moreover using this approach we are able to detect in MS patients a global shift of the T(2) of the white mater toward higher values. The results of this study showed that the method is easy to implement and may be used to characterize MS pathology. PMID- 12591570 TI - Truncation artifact reduction in spectroscopic imaging using a dual-density spiral k-space trajectory. AB - Truncation artifacts arise in magnetic resonance spectroscopic imaging (MRSI) of the human brain due to limited coverage of k-space necessitated by low SNR of metabolite signal and limited scanning time. In proton MRSI of the head, intense extra-cranial lipid signals "bleed" into brain regions, thereby contaminating signals of metabolites therein. This work presents a data acquisition strategy for reducing truncation artifact based on extended k-space coverage achieved with a dual-SNR strategy. Using the fact that the SNR in k-space increases monotonically with sampling density, dual-SNR is achieved in an efficient manner with a dual-density spiral k-space trajectory that permits a smooth transition from high density to low density. The technique is demonstrated to be effective in reducing "bleeding" of extra-cranial lipid signals while preserving the SNR of metabolites in the brain. PMID- 12591571 TI - Three-dimensional numerical simulations of susceptibility-induced magnetic field inhomogeneities in the human head. AB - Three-dimensional numerical simulations of the static magnetic field in the human head were carried out to assess the field inhomogeneity due to magnetic susceptibility differences at tissue interfaces. We used a finite difference method and magnetic permeability distributions obtained by segmentation of computed tomography images. Computations were carried out for four models, consisting of the head and the neck; the head, neck, and shoulders; the head, neck, and thorax; and the head tilted backwards, including the neck and the shoulders. Considerable magnetic field inhomogeneities were observed in the inferior frontal lobes and inferior temporal lobes, particularly near the sphenoid sinus and the temporal bones. Air/tissue interfaces at the shoulders were found to induce substantial magnetic field inhomogeneities in the occipital lobes and the cerebellum, whereas air/tissue interfaces in the lungs appeared to have less influence on the magnetic field in the brain. Tilting the head backwards could significantly reduce the field inhomogeneities superior to the planum sphenoidale as well as in the occipital lobes and the cerebellum. PMID- 12591572 TI - Detection of late epilepsy by the texture analysis of MR brain images in the lithium-pilocarpine rat model. AB - The aim of the study was to detect by texture analysis non easily visible anomalies of magnetic resonance (MR) images of piriform and entorhinal cortices relevant to the lithium-pilocarpine (Li-Pilo) model of temporal lobe epilepsy in rats. Status epilepticus was induced by Li-Pilo in twenty male rats 21 day-old. T(2)-weighted MR images of their brain, were obtained before injection of Li-Pilo and one day after status epilepticus. An hyperintense signal was found in the piriform and entorhinal cortices of six rats, which developed chronic epilepsy after a latent period of one to three months. Among the 14 other rats which displayed images similar to those obtained before injection, four remained healthy but 10 rats developed late epileptic symptoms, raising the problem of hidden cortical damage which may be too subtle to be detected by classic MRI examination. A numeric treatment of digital images was then undertaken by texture analysis, to derive image information from a purely computational point of view. The combined texture and discriminant analyses based on pixels pattern anomalies, selected 3 texture parameters derived from co-occurrence matrix which characterized structural abnormalities relevant to the hyperintense signal, not only in the modified images of 6 rats but also in images of 10 rats with apparently non modified images. These three texture's parameters allowed to classify the twenty rats of our experiment as follows: sixteen epileptic rats were effectively classified with cortical lesions, two non epileptic were correctly classified with healthy cortex, but two healthy rats were not correctly classified. This misclassification is discussed on the basis of the time dependence of the onset of seizure in the Li-Pilo model. These promising results suggest to apply this method to MRI examinations for an improvement of the early diagnostic of human epilepsy. PMID- 12591573 TI - Proton magnetic resonance spectroscopy of the kidney in renal stone disease. AB - Previous studies of renal stone disease (RSD) in Thailand indicated abnormal urinary aggregator and inhibitor composition among farmers with excessive sweat loss. Our aim was to compare the proton MR spectra obtained from the kidneys of 32 proven cases of RSD (aged 38 to 65 yrs) with nine age-matched normal control subjects. We used the STEAM sequence with TE = 15 ms and TR = 2,000 ms. The spectra at 3.25, 3.6 and 3.9 ppm were analyzed. The results showed a correlation between the three peaks (p < 0.001), however, there was no significant difference between the RSD group and the normal control subjects. We therefore concluded that there was no overloading of these osmolytes among the renal stone patients. PMID- 12591574 TI - MRI fast tree log scanning with helical undersampled projection acquisitions. AB - Magnetic Resonance Imaging opens an alternative way to analyze wood structures using a non-destructive technology. It provides high resolution, compound-based contrast manipulation and increased data acquisition flexibility. The technique is particularly useful for tree logs, since they present several characteristics that can be used to reduce the long scan time. This study proposes a method that takes advantage of the log cylindrical symmetry, acquiring transverse 1-D projections with a helical and undersampled pattern. Linear interpolation is used to estimate the skipped data and slice images are reconstructed by filtered backprojection. The sequence is improved using selective multi-pass scanning, without major variations of the scan time. Computer simulations and experimental results show that the proposed technique can increase the scan speed by a factor of 6, while maintaining the ability to identify typical tree log characteristics. PMID- 12591575 TI - Anterior hippocampal volume reduction in male patients with schizophrenia. AB - Quantitative high resolution magnetic resonance imaging (MRI) was utilized to measure anterior, posterior, and total hippocampal volumes in 27 male patients with chronic schizophrenia and 24 male controls. To optimize measurement techniques, hippocampal volumes were: (1) acquired with 1.4-mm slices; (2) excluded with the amygdala; (3) normalized for position; and (4) corrected for total intracranial volume (ICV). The results of a linear mixed effects regression analysis, which made it possible to analyze total anterior and total posterior hippocampal volumes separately, indicated that the anterior hippocampus was significantly smaller in the schizophrenic group relative to the control group. There were no significant group differences with respect to posterior hippocampal volumes, and no significant correlations between hippocampal volumes and illness duration. A significant lateralized asymmetry was also noted in both groups with the right hippocampal volume being larger than the left. These preliminary findings support a significant anterior hippocampal volume reduction in men with schizophrenia as well as a similar hippocampal volume asymmetry in both male controls and schizophrenics. PMID- 12591576 TI - Nerve growth factor in never-medicated first-episode psychotic and medicated chronic schizophrenic patients: possible implications for treatment outcome. AB - Nerve growth factor (NGF) has been found to play a crucial role in the neuroplasticity of predominantly cholinergic neurons in brain development, and neuronal survival following brain injury, which reflect in cognitive performance. Wide ranges of neurodevelopmental abnormalities have been reported in schizophrenic patients, who also show poor cognitive performance. We report plasma NGF levels in never-medicated first-episode psychotic (FEP; N=24) and chronic medicated schizophrenic patients (N=24). NGF levels were determined in plasma by Enzyme-Linked ImmunoSorbent Assay (ELISA). Plasma NGF levels were significantly lower in both FEP and medicated chronic patients as compared to normal subjects (P<0.001). However, NGF levels were significantly higher in chronic schizophrenic patients, which were treated with antipsychotics as compared to FEP (P<0.05). Moreover, NGF levels in chronic patients treated with atypical antipsychotics were markedly higher as compared to patients treated with typical antipsychotics (P<0.05). Lower NGF levels in FEP patients at the onset of psychosis may have implications for the neurodevelopmental abnormalities. However, higher NGF levels in chronic patients treated with atypical antipsychotics may have implications for the treatment outcome. PMID- 12591577 TI - Structural brain abnormalities among relatives of patients with schizophrenia: implications for linkage studies. AB - Several studies suggest that the nonschizophrenic relatives of schizophrenic patients exhibit structural brain abnormalities that may be manifestations of genes that predispose to schizophrenia. In this work, we examine the utility of such measures for linkage analyses. Subjects were 45 nonpsychotic first-degree adult relatives of schizophrenic patients and 48 normal controls. Sixty contiguous 3-mm coronal, T1-weighted 3D magnetic resonance images of the entire brain were acquired on a 1.5-T magnet. We used factor analysis to derive MRI based phenotypes for analysis. The factor analyses produced three factors that significantly discriminated relatives from controls. We used a linear combination of the three factor scores to derive an MRI phenotype. A receiver operating characteristic (ROC) analysis of this phenotype estimated an area under the curve (AUC) statistic of 0.85. The phenotype also discriminated nonpsychotic relatives having two schizophrenic relatives from those having only one. The nonpsychotic relatives of schizophrenic patients show deviant values on MRI measures of brain structure and the distribution of these deviations among relatives and controls suggests that if these results can be replicated, an MRI-derived phenotype could be useful for genetic linkage and association analyses. PMID- 12591578 TI - Volumes of association thalamic nuclei in schizophrenia: a postmortem study. AB - The major association thalamic nuclei, the mediodorsal nucleus (MD) and the medial pulvinar nucleus (PUM) are regarded as important parts of the circuits among association cortical regions. Association cortical regions of the frontal, parietal and temporal lobes have been repeatedly implicated in the neuropathology of schizophrenia. Thus, the aim of the present postmortem study was to investigate the volumes of association thalamic nuclei in this disease. The volumes of the whole thalamus (THAL), MD and PUM were measured in each hemisphere of brains of 12 patients with schizophrenia and 13 age-matched and gender-matched normal control subjects without neuropsychiatric disorders. Patients with schizophrenia exhibited significant volume reductions in both the MD and the PUM, the reductions being more pronounced in the PUM. The volume of the PUM in the left (-19.7%, P=0.02) and right (-22.1%, P=0.01) hemispheres was significantly reduced in the schizophrenia group. The volume of the MD was reduced in both hemispheres in the schizophrenia group. However, the volume reduction was only significant in the left hemisphere (-9.3%, P=0.03). Patients with schizophrenia also exhibited a decreased volume of the THAL in the left (-16.4%, P=0.003) and right (-15.2%, P=0.006) hemispheres. There were no significant correlations between thalamic volumes and duration of illness or age of the patients. In conclusion, the present data indicate volume reductions of association thalamic nuclei in schizophrenia. These anatomical findings are consistent with the view that schizophrenia may be associated with disturbances of association cortical networks. However, the findings of a substantial volume reduction of the THAL suggest that the volumes of additional thalamic nuclei may be also reduced in schizophrenia. PMID- 12591579 TI - A postmortem study of the mediodorsal nucleus of the thalamus in schizophrenia. AB - Four studies have reported that the mediodorsal nucleus of the thalamus (MD) is smaller and contains fewer neurons in schizophrenia. The MD is a key node in a circuit proposed to be dysfunctional in the disorder. However, one study did not find a MD volume loss in schizophrenia, and all the studies to date are relatively small. Given the importance of establishing unequivocally the presence of MD pathology, we have carried out a study of the volume and number of neurons in the left and right MD in 21 patients with schizophrenia and 27 healthy comparison subjects. We also measured the size of MD neurons, and estimated total thalamic volume. We found no difference in the volume of the MD, the number of MD neurons, or the size of MD neurons in either hemisphere in schizophrenia. Neither was total thalamic volume altered. There are no obvious methodological or clinical factors to explain our failure to replicate the finding of MD involvement in schizophrenia. Hence our negative observations, in the largest sample yet investigated, cast doubt on the robustness and/or the generalisability of MD neuropathology in schizophrenia. PMID- 12591581 TI - Morphology of the lateral superior temporal gyrus in neuroleptic nai;ve patients with schizophrenia: relationship to symptoms. AB - OBJECTIVE: The superior temporal gyrus (STG) is a large structure in the temporal lobe with multiple sub-regions that are structurally and functionally distinct. This study evaluates the structural morphology of a specific sub-region of the STG, the anterior and posterior portions of the lateral aspect of the STG. Furthermore, relationships between the morphology of these regions and symptoms of the illness were explored. METHOD: Regions of cortex were consecutively traced on a set of serial coronal slices in 25 male neuroleptic nai;ve patients with first episode schizophrenia and 25 age-matched healthy volunteers. Regional gray matter volumes were calculated and compared, and their correlations with three symptom dimensions were explored. RESULTS: The left anterior STG had a significant inverse correlation with psychotic symptoms, whereas the right posterior STG had a significant positive correlation with negative symptoms. These findings were confirmed by a follow-up analysis using extreme groups. There was no significant correlation between any region and disorganized symptoms. CONCLUSIONS: These findings suggest that abnormalities in the lateral side of the STG may be associated with both psychotic and negative symptoms through different pathophysiological mechanisms. PMID- 12591580 TI - No evidence for association of the dysbindin gene [DTNBP1] with schizophrenia in an Irish population-based study. AB - A recent family-based association study identified a putative association between variants in the dystrobrevin binding protein 1 (dysbindin) gene (DTNBP1) and schizophrenia. This study used a sample of 270 Irish pedigrees multiply affected with schizophrenia. We attempted to replicate these findings in an independent Irish sample of 219 schizophrenia cases and 231 controls. No evidence was found to suggest an association between the DTNBP1 gene and schizophrenia in our sample. Possible reasons for these findings are discussed. PMID- 12591582 TI - Language lateralization in female patients with schizophrenia: an fMRI study. AB - Gender differences in schizophrenia are among the most consistently reported findings in schizophrenia research. However, the biological substrate underlying these gender differences is still largely unknown. Differences in language lateralization between men and women may underlie some gender differences in schizophrenia. In previous functional imaging studies, language lateralization was found to be decreased in male schizophrenia patients as compared to healthy males, which was due to enhanced language activation of the right hemisphere as compared to the healthy males. It could be hypothesized that decreased language lateralization in schizophrenia is gender specific, i.e. decreased lateralization in male patients and normal lateralization in female patients. To test this hypothesis, language activation was measured in 12 right-handed female patients with schizophrenia and 12 healthy females, and compared to findings in 12 male patients and 12 male controls of an earlier study. Language lateralization was significantly lower in the female patients (0.44) as compared to the female controls (0.75), which was due to increased activation of the right-sided language areas (patients: 19 voxels; controls: 8 voxels), while left hemisphere activation was similar in patients and controls. When these data are compared to the male patients and controls, both patient groups had lower lateralization than their healthy counterparts, but there was no difference between male and female patients. In both sexes, decreased lateralization resulted from increased right hemispheric language activation, which suggests a failure to inhibit nondominant language areas in schizophrenia. These findings indicate that lower language lateralization in women is not likely to underlie gender differences in schizophrenia. PMID- 12591583 TI - Morphometric abnormality of the insula in schizophrenia: a comparison with obsessive-compulsive disorder and normal control using MRI. AB - The insula is increasingly the subject of great interest in psychiatric disorders of neurodevelopmental origin because of its anatomical location, wide interconnectivity, and variety of functions. This study explores the possible morphometric change of the insula in schizophrenia and obsessive-compulsive disorder (OCD), and its potential relationship to clinical symptoms. The insula was traced on all coronal slices of magnetic resonance images of three age- and sex-matched diagnostic groups, which consisted of 21 patients with schizophrenia, 21 patients with OCD and 21 normal volunteers. The volumetric measures of the insula were compared among the three groups, and their relationships to the symptom severity were investigated. Volumetric reduction of the left insula was observed in the schizophrenia group, but not in the OCD group. These results confirm the involvement of deficient insular function in the pathophysiology of schizophrenia. PMID- 12591584 TI - Economic evaluations of novel antipsychotic medications: a literature review. AB - OBJECTIVE: To evaluate the evidence that novel antipsychotic medications offer a cost advantage compared to traditional antipsychotic medications. METHODS: Literature for this review was identified through a computerized search of Medline, Healthstar and Psyc-INFO databases inclusive from January 1989 to January 2002. Articles included in the review were required to include cost evaluation and to be published in peer-reviewed journals. RESULTS: Twenty-two studies met inclusion criteria. All five studies that used experimental designs found that second-generation antipsychotic medications were associated with a cost advantage or were cost-neutral, and, in some cases, improved quality of life. Of the ten studies using a pre-post design, four found an increase in total costs, six reported a decrease in total costs, and four reported increased effectiveness with use of a second-generation antipsychotic. All seven of the simulation studies reported a cost advantage for novel antipsychotics for specific patient populations under certain conditions. CONCLUSIONS: The majority of studies found that novel antipsychotics are at least cost-neutral and may offer cost advantages compared to traditional agents. Some studies also reported greater improvement in effectiveness and quality of life when novel antipsychotics were compared to traditional antipsychotic medications. However, it is difficult to draw firm conclusions given the small sample sizes and limited study designs available in this literature. PMID- 12591585 TI - Mental correlates of neuromotoric deviation in 6-year-olds at heightened risk for schizophrenia. AB - The meaning and relevance of the increased rates of neuromotoric deviation (ND) observed in patients with schizophrenia and their biological relatives remain unclear. ND could represent free-floating, independent characteristics of individuals in these families vs. signs of an increased risk for current or future mental disorder. The co-temporaneous relationship between ND and mental disorder at 6 years of age was investigated among 31 children with an increased risk for schizophrenia and similar psychoses, defined as having a mother with a history of schizophrenia or unspecified functional psychosis. As compared with high-risk cases with a low level of ND, the subgroup of 10 high-risk offspring showing notably increased rates of ND had significantly more frequent psychiatric diagnoses (typically language disorders and enuresis), poor functioning on global assessment, poor interpersonal competency and high anxiety proneness. Neuromotoric items representing "overflow" (e.g., choreatic movements, tremor) were significantly positively related to each of these mental characteristics. Among high-risk offspring, an increased rate of ND is very clearly associated with increased rates of current mental disorder, and might potentially identify a subgroup with an especially high risk for serious mental disorder in the future. PMID- 12591586 TI - Abnormalities of motor imagery associated with somatic passivity phenomena in schizophrenia. AB - Some patients with schizophrenia report that their limbs are under the control of an alien force (motor passivity). This is hypothesised to be due to the dysfunction of an internal self-monitoring system that normally permits distinctions between internally generated and external influences on intentional behaviour. Motor imagery is the mental simulation of specific motor actions and it is based upon the internal representation of intended but unexecuted motor actions. Therefore, the generation of motor imagery should be impaired in schizophrenia characterised by passivity phenomena. The generation of motor imagery was compared using the visually guided pointing task (VGPT) and the Florida praxis imagery questionnaire (FPIQ) between patients with schizophrenia characterised by high levels of passivity symptoms (passivity) and patients without passivity symptoms (no-passivity). In both the passivity and no-passivity groups, the speed of real motor sequences on the VGPT was constrained by the distance of the movement and the width of the target in accordance with Fitts' law. For the no-passivity group, the same relationship was found for imagined movements. However, in the passivity group, imagined movements were not constrained by Fitts' law. The effect of a 2-kg load to the limb performing real or imagined movements on the VGPT was identical in both groups. The duration of imagined movements was slowed although the duration of real movements was unaffected. The FPIQ showed that the passivity group had difficulty answering questions that required them to imagine kinaesthetic aspects of performing simple gestures. These results suggest that passivity phenomena in schizophrenia are associated with a specific inability to represent the timing of motor actions internally. This is consistent with the hypothesis that patients with passivity phenomena have difficulty with maintaining an internal representation of intentional behaviour. PMID- 12591587 TI - Childhood developmental abnormalities in schizophrenia: evidence from high-risk studies. AB - According to cohort studies, individuals who develop schizophrenia in adulthood show developmental abnormalities in childhood. These include delays in attainment of speech and motor milestones, problems in social adjustment, and poorer academic and cognitive performance. Another method of investigating developmental abnormalities associated with schizophrenia is the high-risk (HR) method, which follows up longitudinally the development of children at high risk for schizophrenia. Most HR studies have investigated children who have a parent with schizophrenia. This review summarizes findings concerning childhood and adolescent development from 16 HR studies and compares them with findings from cohort, conscript, and family studies. We specifically addressed two questions: (1) Does the development of HR children differ from that of control children? (2) Which developmental factors, if any, predict the development of schizophrenia spectrum disorders in adulthood? While the answer to the first question is affirmative, there may be other mechanisms involved in addition to having a parent with schizophrenia. Factors which appear to predict schizophrenia include problems in motor and neurological development, deficits in attention and verbal short-term memory, poor social competence, positive formal thought disorder-like symptoms, higher scores on psychosis-related scales in the MMPI, and severe instability of early rearing environment. PMID- 12591588 TI - A tri-ethnic examination of symptom expression on the positive and negative syndrome scale in schizophrenia spectrum disorders. AB - This study examined differences in symptom expression as measured by the Positive and Negative Syndrome Scale (PANSS [Schizophr. Bull. 13 (1987) 261]) in a tri ethnic sample of persons diagnosed with schizophrenia. We hypothesized that ethnic differences would be more apparent in Positive Scale symptoms than in Negative and General Scale symptoms of the PANSS. The sample of 351 persons receiving services in community-based mental health clinics came from the initial phase of the San Diego site of the Schizophrenia Care and Assessment Program (SCAP), a longitudinal naturalistic study on the course of schizophrenia treatment. Participants were 88 African-Americans, 198 Euro-Americans, and 65 Latinos. Baseline PANSS scale scores and individual items were analyzed using Multivariate Analysis of Covariance procedures to examine symptoms by ethnic group and living situation while controlling for income, education, and age. There were no significant ethnic differences on the scale scores. At the item level of analysis, significant ethnic group differences were found in Hallucinatory Behavior, Suspiciousness, Excitement, and for Somatic Concerns. The cultural implications for the ethnic differences in each symptom behavior and the need for further research on symptom expression from an ethnographic perspective are discussed. PMID- 12591589 TI - Emotional responses to psychosocial stress in schizophrenia: the role of individual differences in affective traits and coping. AB - Despite the well-established association between psychosocial stress and symptom exacerbation in schizophrenia, factors that account for variability in stress reactivity among individuals with this disorder are unknown. This study examined the association between affective traits, coping style, and neurocognitive functioning and subjective emotional responses during putatively stressful social interactions among individuals with schizophrenia. Self-reported mood was assessed in male schizophrenia outpatients (n=36) and matched nonpsychiatric controls (n=15) during a role-play test (RPT) comprised of simulated social encounters requiring assertive or affiliative skills. During the RPT, schizophrenia patients and controls reported similar elevations in negative mood and decreases in positive mood as compared to baseline mood during assertion scenes. Affiliation scenes resulted only in similar decreases in positive mood across groups as compared to baseline mood. Among schizophrenia patients, trait negative affectivity (NA) and maladaptive coping style accounted for one quarter of the variance in negative mood during the assertion RPTs, and these relationships held after controlling for baseline mood, clinical symptoms, and neurocognitive functioning. Results provide preliminary support for the validity of the social RPT as a paradigm for examining psychosocial stress in schizophrenia and suggest that trait negative affectivity and maladaptive coping are associated with individual differences in emotional responses to psychosocial stressors in schizophrenia. PMID- 12591590 TI - Prefrontal cortex dysfunction during working memory performance in schizophrenia: reconciling discrepant findings. AB - Working memory (WM) deficits are a persistent, disabling and relatively treatment resistant feature of schizophrenia that may underlie many cognitive deficits and symptoms. They are associated with prefrontal cortex dysfunction. While most neuroimaging studies of WM demonstrate "task-related hypofrontality" in schizophrenic relative to healthy subjects, several recent studies have reported equal or increased prefrontal activity. These findings challenge central assumptions regarding cognitive deficits and prefrontal cortex dysfunction in schizophrenia. The goal of this review is to reconcile these seemingly discrepant findings. Methodological factors addressed include the use of intersubject averaging, WM task parameters and the reliability of the measures. Factors intrinsic to schizophrenia and their relevance to the selection of experimental methods and the interpretation of group data are also discussed. Both hypo- and hyperfrontality are hypothesized to be valid and informative reflections of prefrontal cortex dysfunction in schizophrenia. Due to the heterogeneity and variability of both performance and regional recruitment in schizophrenia, whether individual data is considered, the level and type of WM demands and the composition of the sample with regard to performance deficits all influence study outcome and contribute to discrepancies. Although the prefrontal cortex is consistently implicated in WM deficits, the basis of its dysfunction and its exact contribution remain unclear. Future work might focus on delineating the exact WM processes, domains and components that are deficient. In addition, variability in behavior and activation might best be regarded as intrinsic to schizophrenia and having a neural basis that requires explanation. In combination with other techniques, neuroimaging can identify the neural circuitry responsible for WM deficits and elucidate the contribution of each anatomical component. PMID- 12591591 TI - Language comprehension and working memory language comprehension and working memory deficits in patients with schizophrenia. AB - The present study examined the hypothesis that patients with schizophrenia have deficits in language comprehension compared to normal controls, and that these deficits are associated with disturbances in working memory (WM). In addition, we hypothesized that language comprehension deficits would be associated with the severity of specific symptoms in the patients (formal thought disorder and hallucinations). Participants were 27 stable outpatients with schizophrenia and 28 demographically similar controls. Language comprehension was measured by presenting sentences auditorily that varied in length and syntactic complexity, followed by two or three comprehension questions. We measured working memory by administering a reading span task. Results indicated that, as predicted, language comprehension deficits were significantly greater in patients with schizophrenia than controls. Also as predicted, working memory was strongly correlated with language comprehension performance in both patients with schizophrenia and controls. Contrary to our predictions, language comprehension and working memory deficits were not associated with either formal thought disorder or hallucinations. PMID- 12591592 TI - Schizophrenia and tobacco smoking in a Spanish psychiatric hospital. AB - This study in a Spanish hospital replicated two US studies suggesting that schizophrenia is associated with smoking when compared with other severe mental illnesses. Neither antipsychotics nor institutionalism could explain this relationship. Seventy of the 100 schizophrenic and 53 of the 100 non schizophrenic inpatients were current smokers. After correcting for confounding factors, schizophrenia increased the risk of smoking by 2- to 3-fold. Heavy smoking was not associated with schizophrenia. PMID- 12591593 TI - Penultimate observation carried forward (POCF): a new approach to analysis of long-term symptom change in chronic relapsing conditions. PMID- 12591594 TI - Does considering duration of negative symptoms increase their specificity for schizophrenia? PMID- 12591595 TI - Midline brain anomalies in a young schizophrenic patient with 22q11 deletion syndrome. PMID- 12591596 TI - Identification of a novel putative signaling center, the tertiary enamel knot in the postnatal mouse molar tooth. AB - The final shape of the molar tooth crown is thought to be regulated by the transient epithelial signaling centers in the cusp tips, the secondary enamel knots (SEKs), which are believed to disappear after initiation of the cusp growth. We investigated the developmental fate of the signaling center using the recently characterized Slit1 enamel knot marker as a lineage tracer during morphogenesis of the first molar and crown calcification in the mouse. In situ hybridization analysis showed that after Fgf4 downregulation in the SEK, Slit1 expression persisted in the deep compartment of the knot. After the histological disappearance of the SEK, Slit1 expression was evident in a novel epithelial cell cluster, which we call the tertiary enamel knot (TEK) next to the enamel-free area (EFA)-epithelium at the cusp tips. In embryonic tooth, Slit1 was also observed in the stratum intermedium (SI) and stellate reticulum cells between the parallel SEKs correlating to the area where the inner enamel epithelium cells do not proliferate. After birth, the expression of Slit1 persisted in the SI cells of the transverse connecting lophs of the parallel cusps above the EFA-cells. These results demonstrate the presence of a novel putative signaling center, the TEK, in the calcifying tooth. Moreover, our results suggest that Slit1 signaling may be involved in the regulation of molar tooth shape by regulating epithelial cell proliferation and formation of EFA of the crown. PMID- 12591597 TI - Cell-autonomous and signal-dependent expression of liver and intestine marker genes in pluripotent precursor cells from Xenopus embryos. AB - Early regulatory events in respect to the embryonic development of the vertebrate liver are only poorly defined. A better understanding of the gene network that mediates the formation of hepatocytes from pluripotent embryonic precursor cells may help to establish in vitro protocols for hepatocyte differentiation. Here, we describe our first attempts to make use of early embryonic explants from the amphibian Xenopus laevis in order to address these questions. We have identified several novel embryonic liver and intestine marker genes in a random expression pattern screen with cDNA libraries derived from the embryonic liver anlage and from the adult liver of Xenopus laevis. Based on their embryonic expression characteristics, these genes, together with the previously known ones, can be categorized into four different groups: the liver specific group (LS), the liver and intestine group A (LIA), the liver and intestine group B (LIB), and the intestine specific group (IS). Dissociation of endodermal explants isolated from early neurula stage embryos reveals that all genes in the LIB and IS groups are expressed in a cell-autonomous manner. In contrast, expression of genes in the LS and LIA groups requires cell-cell interactions. The regular temporal expression profile of genes in all four groups is mimicked in ectodermal explants from early embryos, reprogrammed by co-injection of VegT and beta-catenin mRNAs. FGF signaling is found to be required for the induction of liver specific marker (LS group) gene expression in the same system. PMID- 12591598 TI - Bruno regulates gurken during Drosophila oogenesis. AB - Translational regulation of localized transcripts is a powerful mechanism to control the precise timing and localization of protein expression within a cell. In the Drosophila germline, oskar transcript must be translationally repressed until its localization at the posterior pole of the oocyte, as ectopic production of Oskar causes severe patterning defects. Translational repression of oskar mRNA is mediated by the RNA-binding protein Bruno, which binds to specific motifs in the oskar 3'UTR. Here we show that Bruno over-expression causes defects in antero posterior and dorso-ventral patterning, consistent with a role of Bruno in both oskar and gurken mRNA regulation. We also show that Bruno and gurken interact genetically. Finally, we show that Bruno binds specifically to the gurken 3'UTR and that the dorso-ventral defects caused by Bruno over-expression are due to a reduction of Gurken levels in the oocyte. We conclude that Bruno plays similar roles in translational regulation of gurken and oskar. PMID- 12591599 TI - Development of adrenal chromaffin cells is largely normal in mice lacking the receptor tyrosine kinase c-Ret. AB - c-Ret encodes a receptor tyrosine kinase that is essential for normal development of the kidney as well as enteric and sympathetic neurons. Since sympathetic neurons and neuroendocrine chromaffin cells originate from a common progenitor cell, we have examined the relevance of c-Ret for the development of adrenal chromaffin cells by analyzing mouse mutants lacking c-Ret. Adrenal chromaffin cells express c-Ret mRNA at embryonic day (E) 12.5 and 13.5, yet levels of expression decline at later embryonic and postnatal ages. Adrenal medullae of c Ret deficient mice show normal numbers of tyrosine hydroxylase (TH) immunoreactive cells at E13.5 and at birth. Ultrastructurally, adrenal chromaffin cells of c-Ret(-/-) mice appear unaltered: chromaffin cells develop typical secretory chromaffin granules, the morphological hallmark of chromaffin cells, and synaptic terminals appear normal. However, adrenaline levels and numbers of chromaffin cells immunoreactive for the adrenaline synthesizing enzyme phenylethanolamine-N-methyltransferase (PNMT) are reduced by about 30% in c-Ret deficient mice arguing for a direct or indirect role of c-Ret in the regulation of PNMT. Thus, despite expression of c-Ret, adrenal chromaffin cells develop largely normal in mice lacking c-Ret. We therefore conclude that sympathetic neurons and neuroendocrine chromaffin cells profoundly differ in their requirement for c-Ret signaling during development. PMID- 12591600 TI - New insights into saccular development and vascular formation in lung allografts under the renal capsule. AB - The study of distal lung morphogenesis and vascular development would be greatly facilitated by an in vitro or ex vivo experimental model. In this study we show that the growth of mouse embryonic day 12.5 lung rudiments implanted underneath the kidney capsules of syngeneic or immunodeficient hosts follows closely lung development in utero. The epithelium develops extensively with both proximal and distal differentiation to the saccular stage. The vasculature also develops extensively. Large blood vessels accompany large airways and capillaries develop within the saccular walls. Interestingly, vessels in the lung grafts develop from endothelial progenitor cells endogenous to the explants and host vessels do not vascularize the grafts independently. This suggests that embryonic lungs possess mechanisms to prevent the inappropriate ingrowth of surrounding vessels. However, vessels in the lung grafts do connect to host vessels, showing that embryonic lungs have the ability to stimulate host angiogenesis and recruit host vessel connections. These data support the hypothesis that the lung vasculature develops by both vasculogenic and angiogenic processes: a vascular network develops in situ in lung mesenchyme, which is then connected to angiogenic processes from central vessels. The lung renal capsule allograft is thus an excellent model to study the development of the pulmonary vasculature and of late fetal lung development that requires a functional blood supply. PMID- 12591601 TI - Loss of XChk1 function triggers apoptosis after the midblastula transition in Xenopus laevis embryos. AB - Prior to the midblastula transition (MBT), Xenopus laevis embryos do not engage cell cycle checkpoints, although overexpression of the kinase XChk1 arrests cell divisions. At the MBT, XChk1 transiently activates and promotes cell cycle lengthening. In this study, endogenous XChk1 was inhibited by the expression of dominant-negative XChk1 (DN-XChk1). Development appeared normal until the early gastrula stage, when cells lost attachments and chromatin condensed. TUNEL and caspase assays indicated these embryos died by apoptosis during gastrulation. Embryos with unreplicated DNA likewise died by apoptosis. Embryos expressing DN XChk1 proceeded through additional rapid rounds of DNA replication but initiated zygotic transcription on schedule. Therefore, XChk1 is essential in the early Xenopus embryo for cell cycle remodeling and for survival after the MBT. PMID- 12591602 TI - The role of a retinoic acid response element in establishing the anterior neural expression border of Hoxd4 transgenes. AB - The zebrafish hoxd4a locus was compared to its murine ortholog, Hoxd4. The sequence of regulatory elements, including a DR5 type retinoic acid response element (RARE) required for Hoxd4 neural enhancer activity, are highly conserved. Additionally, zebrafish and mouse neural enhancers function identically in transgenic mouse embryos. We tested whether sequence conservation reflects functional importance by altering the spacing and sequence of the RARE in the Hoxd4 neural enhancer. Stabilizing receptor-DNA interactions did not anteriorize transgene expression. By contrast, conversion of the RARE from a DR5 to a DR2 type element decreased receptor-DNA stability and posteriorized expression. Hence, the setting of the Hox anterior expression border is not a simple function of the affinity of retinoid receptors for their cognate element. PMID- 12591603 TI - Redundant early and overlapping larval roles of Xsox17 subgroup genes in Xenopus endoderm development. AB - We have used antisense morpholino oligos to establish the developmental roles of three Xsox17 proteins in Xenopus development (Xsox17alpha(1), alpha(2) and beta). We show that their synthesis can be inhibited with modest amounts of oligo. The inhibition of each individually produces defects in late midgut development. Loss of activity of the Xsox17alpha proteins additionally inhibits hindgut formation, and inhibiting Xsox17alpha(1) disrupts foregut development with variable penetrance. When all Xsox17 activity is inhibited cell movements are halted during late gastrulation and the transcription of several endodermally expressed genes is reduced. Thus the Xsox17 proteins have redundant roles in early development of the endoderm and partly distinct roles during later organogenesis. PMID- 12591604 TI - Developmental cooperation of leukemia inhibitory factor and insulin-like growth factor I in mice is tissue-specific and essential for lung maturation involving the transcription factors Sp3 and TTF-1. AB - The multifunctional proteins leukemia inhibitory factor (LIF) and insulin-like growth factor I (IGF-I) are expressed in overlapping patterns during development and, therefore, may act cooperatively. We show that mice doubly deficient in LIF and IGF-I all died at birth of apparent respiratory failure. Growth retardation, muscle hypoplasia and delayed ossification in IGF-I-deficient E18.5 mice were exacerbated by the absence of LIF. The transcription factor Sp3 was decreased in the skeleton of the double null mice. Pronounced depletion of olfactory bulb neurons, in contrast, was only IGF-I-dependent. The lungs displayed reduced air space in the IGF-I-deficient embryos and neonates, phenotype exacerbated in the double nulls, which showed abnormal epithelial cells and decreased Sp3 expression. In addition, the transcription factor TTF-1 and the surfactant protein B were lower in the lung of the double null neonates than in all other genotypes. LIF and IGF-I, thus, have cooperative and distinct tissue functions during development. Their essential role in bone ossification apparently involves Sp3, and in lung maturation Sp3 together with TTF-1. PMID- 12591605 TI - Retinoic acid affects gene expression and morphogenesis without upregulating the retinoic acid receptor in the ascidian Ciona intestinalis. AB - Many chordate-specific morphological features develop depending on retinoic acid (RA). We isolated cDNA clones encoding a retinoic acid receptor (CiRAR) and a retinoid X receptor (CiRXR) in the ascidian Ciona intestinalis. CiRAR mRNA was detected in the anterior ectoderm and endoderm during gastrulation. The expression persists in the head endoderm and two discrete regions of the nerve cord in the tailbud embryo. CiRXR mRNA was ubiquitously expressed. RA affected closure of the neural tube and formation of the adhesive papillae. However, no obvious upregulation in CiRAR expression was observed. Expression of some, but not all, of the neural and papilla-specific genes was reduced in the RA-treated embryo. These results suggest limited roles of CiRAR in ascidian embryos. PMID- 12591606 TI - found in neurons, a third member of the Drosophila elav gene family, encodes a neuronal protein and interacts with elav. AB - elav, a gene necessary for neuronal differentiation and maintenance in Drosophila, encodes the prototype of a family of conserved proteins involved in post-transcriptional regulation. We identified found in neurons (fne), a gene encoding a new ELAV paralogue. We showed that FNE binds RNA in vitro. fne transcripts are present throughout development and contain long untranslated regions. Transcripts and proteins are restricted to neurons of the CNS and PNS during embryogenesis. These features are reminiscent of elav. However, fne expression is delayed compared to elav's, and FNE protein appears cytoplasmic, while ELAV is nuclear. GAL4-directed overexpression of fne in neurons leads to a reduction of stable transcripts produced from both the fne and elav endogenous loci, suggesting that fne autoregulates and also regulates elav. PMID- 12591607 TI - Identification and characterization of a novel mouse plexin, plexin-A4. AB - Plexins belonging to the plexin-A subfamily form complexes with neuropilins and propagate signals of class 3 semaphorins into neurons, even though they do not directly bind the semaphorins. In this study, we identified a new member of the plexin-A subfamily in the mice, plexin-A4, and showed that it was expressed in the developing nervous system with a pattern different to that of other members of the plexin-A subfamily (plexin-A1, plexin-A2 and plexin-A3). COS-7 cells coexpressing plexin-A4 with neuropilin-1 were induced to contract by Sema3A, a member of the class 3 semaphorin. Ectopic expression of plexin-A4 in mitral cells that are originally insensitive to Sema3A resulted in the collapse of growth cones in the presence of Sema3A. These results suggest that plexin-A4 plays a role in the propagation of Sema3A activities. PMID- 12591609 TI - Molecular characterization and expression of equine testicular cytochrome P450 aromatase. AB - We characterized testicular equine aromatase and its expression. A 2707 bp cDNA was isolated, it encoded a polypeptide of 503 residues with a deduced molecular mass of 57.8 kDa. The sequence features were those of a cytochrome P450 aromatase, with a 78% polypeptide identity with the human counterpart. The gene has a minimal length of 74 kb comprising at least 9 exons and expresses a 2.8 kb mRNA in the testis. Transient cDNA transfections in E293 cells and in vitro translations in a reticulocyte lysate system allowed aromatase protein and activity detections. The activity increased with androstenedione as substrate in a dose-dependent manner. The isolation of testicular aromatase by a new immunoaffinity method demonstrated that the protein could exist either glycosylated or not with a 2 kDa difference. All these results taken together allow new structural studies to progress in the understanding of this cytochrome P450. PMID- 12591610 TI - "Action-at-a distance" of a new DNA oxidative damage product 6-furfuryl-adenine (kinetin) on template properties of modified DNA. AB - N(6)-furfuryladenine (kinetin, K) was shown to have cytokinin activity and antiageing effects. It also appears to protect DNA against oxidative damage mediated by the Fenton reaction. Kinetin was identified as a natural component of DNA in plant extract, calf thymus DNA, fresh DNA preparations from human cell culture, as well as in human urine. A proposed mechanism of kinetin synthesis includes furfural, the oxidative damage product of a 2-deoxyribose moiety of DNA, which reacts with an adenine residue to form N(6)-furfuryladenine at DNA level. The identification of kinetin in plant cell extracts, as well as human urine, suggests its excision from DNA by repair mechanisms. Since such a bulky modification as kinetin induces conformational changes of DNA, this could lead to mutations. Therefore, it was interesting to analyze an effect of kinetin on coding properties of DNA. Chemically synthesized oligodeoxynucleotide (20-mer) containing kinetin AAAACTGCCGTCCTGAKGAT was used as a primer. It was elongated in a polymerase chain reaction (PCR) on a template plasmid pEW1 harboring a 210-bp fragment of DNA derived from the 5' end of HIV mRNA. The PCR product of that length containing kinetin in position 17 from the 5' end was isolated and sequenced. Interestingly, DNA polymerase correctly incorporates thymine opposite of kinetin (an adenine derivative) on the complementary strand, but the misincorporations occur in a vicinity of the modified base. PMID- 12591611 TI - Structure and promoter activity of the human glia maturation factor-gamma gene: a TATA-less, GC-rich and bidirectional promoter. AB - Human glia maturation factor-gamma (hGMFG) was recently identified as a gene that is homologous to glia maturation factor-beta (GMFB). In this study, we determined the organization of the 9.5-kb hGMFG gene and characterized its promoter activity. The 5'-flanking region of the first exon has putative elements for binding transcription factors Sp-1, GATA-1, AML-1a, Lyf-1 and Ets-1, but there were no TATA or CAAT boxes within a 226-bp sequence upstream from the initiation codon. Primer extension analysis and 5'RACE (rapid amplification of cDNA 5' ends) identified multiple transcription initiation sites within the region -84 to -70 nucleotides from the first ATG codon in a Kozak consensus sequence. A core promoter region was determined by transfecting a series of deletion constructs with a dual luciferase reporter system into rat astrocyte-derived ACT-57 cells. We found that 226 bp of the core promoter region exhibited bidirectional promoter activity. PMID- 12591612 TI - Molecular characterization of three 3-hydroxy-3-methylglutaryl-CoA reductase genes including pathogen-induced Hmg2 from pepper (Capsicum annuum). AB - Sesquiterpene phytoalexins, a class of plant defense metabolites, are synthesized from the cytosolic acetate/mevalonate pathway in isoprenoids biosynthetic system of plants. The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the synthesis of mevalonate, which is the specific precursor of this pathway, as a multi gene family. Three kinds of cDNA clones encoding HMGR were isolated from Korean red pepper (Capsicum annuum L. cv. NocKwang) and the HMGR2 gene (Hmg2) was especially obtained from a cDNA library constructed with Phytophthora capsici infected pepper root RNAs. The Hmg2 encoding a 604-amino-acid peptide had typical features as an elicitor-induced isoform among HMGRs on its gene structure and had a predicted amino acid sequence homology. In addition, the expression of Hmg2 was rapidly induced within 1 h in response to a fungal pathogen and continuously increased up to 48 h. Together with sesquiterpene cyclase gene that was strongly induced 24 h after pathogen-infection, the Hmg2 and farnesyl pyrophosphate synthase gene were coordinately and sequentially regulated for the biosynthesis of defense-related sesquiterpene phytoalexins in pepper. PMID- 12591613 TI - Identification and characterization of two new members of the GRAS gene family in rice responsive to N-acetylchitooligosaccharide elicitor. AB - We identified two new members of the GRAS gene family from rice, CIGR1 and CIGR2, which are rapidly induced upon N-acetylchitooligosaccharide elicitor perception. The predicated proteins encoded by CIGR1 and CIGR2 possess significant sequence similarity with previously identified members of the GRAS family, such as Arabidopsis SCARECROW, GAI, RGA, tomato Lateral suppressor, and rice SLR1, all of which have VHIID regions, likely to play a role in cellular signaling. Fusions of CIGR1 and CIGR2 with Green Fluorescent Protein were detected exclusively in the nuclei of onion epidermal cells. The expression of CIGR1 and CIGR2 was dependent on the structure of N-acetylchitooligosaccharides, which parallels the structural specificity for chitin binding to the plasma membrane-localized chitin-binding protein, and independent of de novo protein synthesis. Co-cultivation of rice cells with rice blast fungus strongly induced the expression of CIGR1 and CIGR2, whereas inoculation of suspension cells with phytopathogenic bacteria did not. We hypothesize that CIGR1 and CIGR2 act as transcriptional regulators in the early events of the elicitor-induced defense response in rice. PMID- 12591614 TI - Molecular cloning, differential expression, and functional characterization of a family of class I ubiquitin-conjugating enzyme (E2) genes in cotton (Gossypium). AB - Two cDNAs and their corresponding genes (GhUBC1 and GhUBC2) encoding ubiquitin conjugating enzymes (E2s) have been cloned and characterized from allotetraploid cotton Gossypium hirsutum ((AD)(1) genome). Three additional E2 genes (GaUBC1, GtUBC2, and GrUBC2) have also been identified from diploid cottons Gossypium arboreum (A(2) genome), Gossypium thurberi (D(1) genome), and Gossypium raimondii (D(5) genome), respectively. The derived amino acid sequences of the five closely related cotton E2s are 77-79% identical to yeast ScUBC4 and ScUBC5. The GhUBC1/2 gene family is composed of two members, and genomic origin analysis indicates that GhUBC1 and 2 are individually present in the A and D subgenomes of G. hirsutum. The transcript levels of GhUBC1/2 increased significantly in leaves and flowers at senescence, suggesting that GhUBC1/2 may play a role in the degradation of target proteins that function in the delay of the senescence program. Correlated with high auxin content and auxin-associated effects, GhUBC1/2 are also highly expressed in the youngest leaves, the apical part of lateral roots, and elongating fibers. Genetic complementation experiments revealed that GhUBC1 and 2 can substitute for the function of ScUBC4 and 5 required for the selective degradation of abnormal and short-lived proteins in a yeast ubc4ubc5 double mutant. PMID- 12591615 TI - Regulation of renin enhancer activity by nuclear factor I and Sp1/Sp3. AB - Transcription of the mouse Ren-1(c) gene in kidney tumor-derived As4.1 cells, which express high levels of renin mRNA, is dependent on a proximal promoter element and a 242-bp enhancer region located 2.6 kb upstream of the transcription start site. We showed previously that the enhancer contains a cAMP responsive element (CRE) and an E-box. Mutation of either element resulted in almost complete loss of the Ren-1(c) expression. In this report we show that there are additional transcription factor-binding sites within the Ren-1(c) enhancer contributing to the enhancer activity. Electrophoretic mobility shift and supershift assays have identified four nuclear factor I (NFI)-binding sites, an Sp1/Sp3 site and an unidentified transcription factor-binding site (Ei) located upstream of the CRE and E-box. Mutation of the Sp1/Sp3 site or Ei reduced Ren 1(c) expression by 40% or 30%, respectively, while mutations of four NFI-binding sites resulted in an 89% decrease in expression. Thus, these protein-DNA interaction sites are essential for transcription of mouse renin genes. There are four homologous NFI genes (NFI-A, -B, -C and -X) in vertebrates and multiple alternatively spliced isoforms from each gene. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) assays have demonstrated that NFI-X is the predominant NFI mRNA expressed in As4.1 cells. Direct study of involvement of NFI X in regulation of renin genes is underway. PMID- 12591616 TI - Identification of human dehydrodolichyl diphosphate synthase gene. AB - We isolated a cDNA encoding human dehydrodolichyl diphosphate (Dedol-PP) synthase and expressed the gene in a yeast mutant strain SNH23-7D, which is deficient in Dedol-PP synthase activity. The identity of the cloned enzyme was confirmed by functional complementation of SNH23-7D strain together with in vitro Dedol-PP synthase activity assay. Northern blot analysis indicated that testis and kidney expressed Dedol-PP synthase mRNA at high levels. PMID- 12591617 TI - Molecular cloning and characterization of a novel dual-specificity phosphatase18 gene from human fetal brain. AB - Dual-specificity protein phosphatases (DSPs), a new family of protein tyrosine phosphatases (PTPs), are characterized by the ability to dephosphorylate both phospho-tyrosyl and phospho-seryl/threonyl residues. It has been known that most of the enzymes play important roles in the regulation of mitogenic signal transduction and control the cell cycle in response to extracellular stimuli. In this study, a novel human DSP gene named Dual-specificity Phosphatase18 (DUSP18) was isolated by large-scale sequencing analysis of a human fetal brain cDNA library. DUSP18 is localized at Chromosome 22 q12.1. Its cDNA is 2450 base pairs in length, encoding a 188-amino acid polypeptide in which a DSP motif but not a CH2 domain is included. RT-PCR revealed that the DUSP18 was widely expressed in different tissues. GST-DUSP18 fusion protein showed distinctive phosphatase activity toward p-nitrophenyl phosphate (pNPP), as well as oligopeptides containing pThr and pTyr, indicating that DUSP18 is a protein phosphatase with dual substrate specificity. The optimal condition for the reaction was pH 6.0 and 55 degrees C. Addition of Mn(2+) ions was able to enhance the enzyme activity while the activity was strongly inhibited by iodoaretic acid. Mutations in selected sites showed the importance of Asp-73, Cys-104, Arg-110 and Ser-111 in phosphatase activity of DUSP18. PMID- 12591618 TI - cDNAs for the synthesis of cyclic carotenoids in petals of Gentiana lutea and their regulation during flower development. AB - cDNAs encoding lycopene epsilon -cyclase, lycopene beta-cyclase, beta-carotene hydroxylase and zeaxanthin epoxidase were isolated from a Gentiana lutea petal cDNA library. The function of all cDNAs was analyzed by complementation in Escherichia coli. Transcript levels during different stages of flower development of G. lutea were determined and compared to the carotenoid composition. Expression of all genes increased by a factor of up to 2, with the exception of the lycopene epsilon -cyclase gene. The transcript amount of the latter was strongly decreased. These results indicate that during flower development, carotenoid formation is enhanced. Moreover, metabolites are shifted away from the biosynthetic branch to lutein and are channeled into beta-carotene and derivatives. PMID- 12591619 TI - A qualitative analysis of a model for the transmission of varicella-zoster virus. AB - Varicella-zoster virus (VZV) is a herpesvirus which is the known agent for causing varicella (chickenpox) in its initial manifestation and zoster (shingles) in a reactivated state. The standard SEIR compartmental model is modified to include the cycle of shingles acquisition, recovery, and possible reacquisition. The basic reproduction number R(0) shows the influence of the zoster cycle and how shingles can be important in maintaining VZV in populations. The model has the typical threshold behavior in the sense that when R(0)1, the virus persists over time and so chickenpox and shingles remain endemic. PMID- 12591620 TI - Prostate cancer: progression of prostate-specific antigen after external beam irradiation. AB - This paper is concerned with the development of a stochastic path of prostate specific antigen (PSA) level after radiation treatment for prostate cancer. PSA is a biomarker for prostate cancer, higher levels of which indicate the seriousness of the cancer progression. Following the deterministic modeling of the data by the previous authors, Cox et al., this paper is concerned with the theoretical knowledge that could be gained by the stochastic modeling in discrete form of the PSA path over time. The expected value of the PSA level is computed and compared with the deterministic model and it is found that they are the same for about the first year after radiation therapy. The American Society for Therapeutic Radiology has set a consensus panel definition of biochemical failure following radiation therapy: the rise in three consecutive levels of PSA is considered to be a failure of the radiation therapy. Knowledge of the path of PSA presented in this paper would be useful in the management of the radiation treatment and in particular assessing quantitatively any clinically based policy for defining recurrence after radiation therapy. Application of the model is illustrated by fitting it to clinical data available in the University of Michigan cancer center. PMID- 12591621 TI - Exploiting thermal noise for an efficient actomyosin sliding mechanism. AB - With reference to the experimental observations by Yanagida and his co-workers concerning actomyosin interaction during muscle contraction processes, we propose a phenomenological model for the sliding of the myosin head on the actin filament, in which the myosin head is viewed as an active Brownian particle in a periodic, elastic-type potential subject to tilting. The sample paths thus obtained are qualitatively alike to those experimentally recorded. Furthermore, our model is proved to be susceptible of a consistent parameters regulation yielding step frequencies, mean step dwell time and dwell time distribution in excellent agreement with the experimental evidence. PMID- 12591622 TI - Multiple limit cycles in the chemostat with variable yield. AB - The global asymptotic behavior of solutions of the variable yield model is determined. The model generalizes the classical Monod model and it assumes that the yield is an increasing function of the nutrient concentration. In contrast to the Monod model, it is demonstrated that the variable yield model exhibits sustained oscillations. Moreover, it is shown that the variable yield model may undergo a subcritical Hopf bifurcation and feature at least two distinct limit cycles. Implications for the coexistence of competing populations are discussed. PMID- 12591623 TI - A new family of global protein shape descriptors. AB - A family of global geometric measures is constructed for protein structure classification. These measures originate from integral formulas of Vassiliev knot invariants and give rise to a unique classification scheme. Our measures can better discriminate between many known protein structures than the simple measures of the secondary structure content of these protein structures. PMID- 12591624 TI - A note on estimation of dynamics of multiple gene expression based on singular value decomposition. AB - Recently, data on multiple gene expression at sequential time points were analyzed, using singular value decomposition (SVD) as a means to capture dominant trends, called characteristic modes, followed by fitting of a linear discrete time dynamical system in which the expression values at a given time point are linear combinations of the values at a previous time point. We attempt to address several aspects of the method. To obtain the model we formulate a non-linear optimization problem and present how to solve it numerically using standard MATLAB procedures. We use publicly available data to test the approach. For reader's convenience, we provide a straightforward, ready-to-use, procedure in MATLAB, which employs its standard features to analyze data of this kind. Then, we investigate the sensitivity of the method to missing measurements and its possibilities to reconstruct missing data. Also, we discuss the possible consequences of data regularization, called sometimes 'polishing', on the outcome of analysis, especially when model is to be used for prediction purposes. Summarizing we point out that approximation of multiple gene expression data preceded by SVD provides some insight into the dynamics but may also lead to unexpected difficulties, like overfitting problems. PMID- 12591625 TI - A differentiable, periodic function for pulsatile cardiac output based on heart rate and stroke volume. AB - Many mathematical models of human hemodynamics, particularly those which describe pressure and flow pulses throughout the circulatory system, require as specified input a modeling function which describes cardiac output in terms of volume per unit time. To be realistic, this cardiac output function should capture, to the greatest extent possible, all relevant features observed in measured physical data. For model analysis purposes, it is also highly desirable to have a model function that is continuous, differentiable, and periodic. This paper addresses both classes of needs by developing such a function. Physically, the present function provides an accurate model for flow into the ascending aorta. It is completely specified by a minimal number of standard input parameters associated with left ventricle dynamics, including heart rate, mean cardiac output, and an estimation of the peak-to-mean flow ratio. Analytically, it can be expressed as a product of two continuous, differentiable and periodic factors. Further, the Fourier expansion of this model function is shown to be a finite Fourier series, and explicit closed-form expressions are given for the non-zero coefficients in this series. PMID- 12591626 TI - Integrals for continuous-time Markov chains. AB - This paper presents a method of evaluating the expected value of a path integral for a general Markov chain on a countable state space. We illustrate the method with reference to several models, including birth-death processes and the birth, death and catastrophe process. PMID- 12591627 TI - MR identification of Chiari pathophysiology by using spatial and temporal CSF flow indices and implications for syringomyelia. PMID- 12591628 TI - New model of minimally invasive experimental spinal cord injury. PMID- 12591629 TI - Peak systolic and diastolic CSF velocity in the foramen magnum in adult patients with Chiari I malformations and in normal control participants. AB - BACKGROUND AND PURPOSE: Abnormal flow of CSF through the foramen magnum has been implicated in the pathogenesis of clinical deficits in association with Chiari I malformation. The purpose of this study was to test the hypothesis that peak CSF velocities in the foramen magnum are increased in patients with Chiari I malformations. METHODS: Eight adult patients with symptomatic Chiari I malformations and 10 adult volunteers were studied with cardiac gated, phase contrast MR imaging in the axial plane at the foramen magnum. The spatial uniformity of flow velocity in the foramen magnum was assessed at 14 time frames within the R-R interval. The velocity in each of the voxels at each of the time frames was calculated, and the peak systolic and diastolic velocities were tabulated for the patients and controls. RESULTS: For the normal volunteers, the CSF velocities in the subarachnoid space were relatively uniform throughout the subarachnoid space at each of the time frames. Peak systolic velocity ranged from 1.2 to 3.3 cm/s, and peak diastolic velocity ranged from 1.6 to 4.5 cm/s. In symptomatic patients with Chiari I, velocities in the foramen magnum did not appear as uniform throughout the subarachnoid space in the phase-contrast images. Peak systolic velocities ranged from 1.8 to 4.8 cm/s, and peak diastolic velocities ranged from 2.5 to 5.3 cm/s. Peak systolic velocity was significantly higher (P =.01) in the patients than in the control volunteers. CONCLUSION: Patients with Chiari I malformations have significant elevations of peak systolic velocity in the CSF in the foramen magnum. PMID- 12591631 TI - Increase in vertebral body height after vertebroplasty. AB - BACKGROUND AND PURPOSE: During clinical work, we have seen increases in vertebral body height associated with vertebroplasty, but our literature search revealed no reports as to how often and to what degree those increases occur. The purpose of this study was to document the frequency and degree of changes in vertebral body height after vertebroplasty. METHODS: The heights of 85 vertebral bodies in 37 patients were measured before and after vertebroplasty. In addition, one adjacent vertebral body was measured in each patient as a control. Twenty-six patients had compression fractures in the thoracic spine, and 24 patients had compression fractures in the lumbar spine. Vertebroplasty was performed with a bilateral transpedicular approach by injecting polymethylmethacrylate, under biplane fluoroscopic control. Measurements were performed on preoperative MR images and on postoperative CT sagittal reformations. Anterior, central, and posterior vertebral body height measurements were obtained in the midsagittal plane. RESULTS: The average increase in vertebral body height was 2.5 mm anteriorly, 2.7 mm centrally, and 1.4 mm posteriorly. Thirteen of 85 treated vertebrae remained unchanged. All control vertebral bodies also remained unchanged. CONCLUSION: Vertebral body height often increases during vertebroplasty. The clinical significance of increasing vertebral body height is unknown. PMID- 12591630 TI - Percutaneous translumbar spinal cord compression injury in a dog model that uses angioplasty balloons: MR imaging and histopathologic findings. AB - BACKGROUND AND PURPOSE: Previous animal models for spinal cord injury required laminectomy and exposure of the spinal cord to create direct trauma, compromising imaging by both surgical artifact and the nature of the production of the injury. Our purpose was to study a model that uses percutaneous intraspinal navigation with an angioplasty balloon, providing a controlled degree of spinal cord compression and allowing improved MR imaging of spinal cord injury. METHODS: Nine mongrel dogs were studied. MR images were obtained of six dogs after technique development in three dogs. Angioplasty balloons measuring 7 or 4 mm in diameter and 2 cm in length were placed in the midthoracic subarachnoid space. Imaging was performed by using a 1.5-T MR imaging unit before and after balloon inflation. The balloon was inflated within 5 seconds and deflated after 30 minutes. T1- and T2-weighted and contrast-enhanced images were acquired. Spinal cords were submitted for pathologic examination. RESULTS: All four animals with 7-mm balloons experienced hemorrhage, and three had axonal injury revealed by histopathologic examination. One of two animals with 4-mm balloons experienced no injury, and one had axonal injury without hemorrhage. Regional parenchymal enhancement was seen in two of the animals with 7-mm balloons. CONCLUSION: This percutaneous spinal cord injury model results in a graduating degree of injury. It differs from previous techniques by avoiding surgical exposure and the associated artifacts, yet it offers histopathologic findings similar to those of human spinal cord injury. The canine spinal cord is amenable to MR imaging with clinical imaging units. Further evaluations with various durations of compression and various balloon sizes are warranted. PMID- 12591632 TI - Dumbbell-shaped epidural capillary hemangioma. AB - We report a case of a purely epidural capillary hemangioma of the thoracic spine with foraminal extension. Epidural hemangiomas are rare; only a few cases of dumbbell-shaped ones have been reported, and all were cavernous. MR imaging showed characteristic findings of a capillary hemangioma, which are also consistent with other epidural lesions such as neuromas or meningiomas. PMID- 12591633 TI - Quantity determination and the distance effect with letters, numbers, and shapes: a functional MR imaging study of number processing. AB - BACKGROUND AND PURPOSE: The ability to quantify, or to determine magnitude, is an important part of number processing, and the extent to which language and other cognitive abilities are involved with number processing is an area of interest. We compared activation patterns, reaction times, and accuracy as subjects determined stimulus magnitude by ordering letters, numbers, and shapes. A second goal was to define the brain regions involved in the distance effect (the farther apart numbers are, the faster subjects are at judging which number is larger) and whether this effect depended on stimulus type. METHODS: Functional MR images were acquired in 19 healthy subjects. The order task required the subjects to judge whether three stimuli were in order according to their position in the alphabet (letters), position in the number line (numbers), or size (shapes). In the control (identify task), subjects judged whether one of the three stimuli was a particular letter, number, or shape. Each stimulus type was divided into near trials (quantity difference of three or less) and far trials (quantity difference of at least five) to assess the distance effect. RESULTS: Subjects were less accurate and slower with letters than with numbers and shapes. A distance effect was present with shapes and numbers, as subjects ordered the near trials slower than far trials. No distance effect was detected with letters. The occipital lobes and intraparietal sulci were active with all three stimuli. Shapes required no additional areas, although analysis of the distance effect revealed that near shapes involved other brain regions, including the frontal lobes. Letters activated a large network comprising the frontal lobes, the anterior cingulate gyrus, and basal ganglia, but few additional areas were engaged by the distance effect. Numbers involved a smaller network including the inferior and middle frontal gyri. The left supramarginal gyrus and both inferior frontal gyri were active in the distance effect with numbers. CONCLUSION: Numbers and letters, which are stimuli more associated with abstract symbolism compared with shapes, resulted in slower reaction times and an increased number of active brain regions. Shapes and numbers, but not letters, elicited a distance effect, indicating access to a mental continuum of quantity. The left supramarginal gyrus and both inferior frontal gyri were involved in the distance effect with numbers. The intraparietal sulci were important in determining stimulus magnitude for all stimuli. PMID- 12591634 TI - Functional MR imaging of the auditory cortex with electrical stimulation of the promontory in 35 deaf patients before cochlea implantation. AB - BACKGROUND AND PURPOSE: Promontory testing is used for preoperative assessment of the auditory pathway before cochlear implantation. This method depends on patient cooperation and cannot be used in children or disabled persons. Promontory stimulation during functional MR imaging (fMRI) provides a new and objective method to test the integrity of the auditory pathway. To evaluate the method, we performed this prospective study in deaf adult patients. METHODS: fMRI of the auditory pathway with electrical stimulation of the promontory was performed in 35 profoundly deaf patients, bilaterally in seven. For safe stimulation inside the MR environment, a specially designed nerve stimulator was used. We acquired nine sections parallel to the sylvian fissure by using an echo-planar pulse sequence (1.5 T). To evaluate the number of pixels in the auditory cortex, areas were counted and the minimum confidence level (p(st) value) was determined. The auditory pathway was called intact when the minimal p(st) value was 10(-5) or when the minimal p(st) value was 10(-4) in at least five activated pixels. RESULTS: Images in 85% of patients reporting an auditory sensation showed activation of the contralateral auditory cortex. In the group of patients reporting no hearing sensation, images in 75% did not show activation. CONCLUSION: This method can prove the intactness of the auditory pathway and help the surgeon in decision making before cochlear implantation. However, a negative finding should not be interpreted as indicating a nonfunctioning auditory pathway. Additional technical refinements and experience are needed to further improve this method. PMID- 12591635 TI - Role of the corpus callosum in functional connectivity. AB - BACKGROUND AND PURPOSE: Regional cerebral blood flow fluctuates synchronously in corresponding brain regions between the hemispheres. This synchrony implies neuronal connections between brain regions. The synchrony of blood flow changes is defined operationally as functional connectivity. Our purpose was to measure functional connectivity in patients with corpus callosal agenesis, in whom the interhemispheric connectivity is hypothetically diminished. METHODS: In three patients with agenesis of the corpus callosum, functional MR imaging was performed while patients performed text-listening and finger-tapping tasks. Functional images were also acquired while the patients performed no specific task (resting state). Regions of activation temporally correlated with the performance of the tasks were identified by cross-correlating the task data with a reference function. Voxel clusters (seed voxels) that corresponded to regions of activation in the task-activation data set were selected in the resting data set. All the voxels in the resting 3D data set that had a correlation coefficient exceeding 0.4 were identified. The number of these voxels in the ipsilateral and contralateral hemispheres was tabulated. RESULTS: In all patients, technically adequate functional MR and functional connectivity MR maps were obtained. For both tasks, activation was found in both hemispheres. For all of the seed voxels, significantly more functionally connected voxels were found in the ipsilateral hemisphere than in the contralateral hemisphere. For most seed voxels, no functionally connected voxels were found in the contralateral hemisphere. CONCLUSION: Interhemispheric functional connectivity in the motor and auditory cortices is diminished in patients with agenesis of the corpus callosum compared with that of healthy subjects. PMID- 12591636 TI - Pseudo-reorganization of language cortical function at fMR imaging: a consequence of tumor-induced neurovascular uncoupling. AB - A left-handed patient with a grade II left frontal lobe astrocytoma had spontaneous seizures causing speech arrest and uncontrolled right upper extremity movements. Word-generation functional MR (fMR) imaging showed activity nearly exclusively in the right inferior frontal gyrus. The clinical history of the speech arrest and the intraoperative mapping proved left-hemisphere language dominance. Tumor involvement of the left inferior frontal gyrus caused uncoupling of the blood oxygenation level-dependent (BOLD) and neuronal response, leading to the erroneous fMR imaging appearance of right-hemisphere language dominance. Discrepancies between BOLD and intraoperative mapping in areas near lesions illustrate the complementary nature of these techniques. PMID- 12591637 TI - Magnetization transfer MR imaging in patients with posttraumatic epilepsy. AB - BACKGROUND AND PURPOSE: Intractable epilepsy is a well-recognized complication following head trauma, and many factors have been implicated in its pathogenesis. This study was performed to determine the severity of tissue damage after severe head injury as assessed with magnetization transfer (MT) MR imaging and the relationship of this damage with seizure intractability. METHODS: Forty-four patients, 13 without seizures (disease controls) and 31 with seizures, underwent T1-weighted MT MR imaging 1-10 years after head trauma. Phase-corrected gradient echo (GRE) imaging was also performed in all patients to look for the presence of hemosiderin. All patients were evaluated for the presence of an MT abnormality beyond an abnormality seen on T2-weighted images, an MT abnormality within a T2 abnormality, and hemosiderin deposition. RESULTS: Patients with an MT abnormality beyond a T2 abnormality had a significantly higher intractability of seizures compared with those with an MT abnormality within a T2 abnormality (P <.05). In addition, the mere presence of hemosiderin deposit was not associated with seizure intractability; however, gliosis around the hemosiderin as seen on T1 weighted MT images was associated with seizure intractability. CONCLUSIONS: T1 weighted MT imaging may be of value in predicting the intractability of the seizure in delayed posttraumatic epilepsy. PMID- 12591638 TI - Combination of single-voxel proton MR spectroscopy and apparent diffusion coefficient calculation in the evaluation of common brain tumors. AB - BACKGROUND AND PURPOSE: MR spectroscopy and apparent diffusion coefficient (ADC) calculation have been used frequently for tumor grading and differentiation during the last decade. We evaluated whether the combination of these two techniques can improve the diagnostic effectiveness of MR imaging in patients with brain tumors. METHODS: Forty-nine patients with histologically proved brain tumors (eight high- and 12 low-grade astrocytomas, eight metastases, eight nonastrocytic gliomas, seven meningiomas, three dysembryoplastic neuroepithelial tumors (DNETs), and three tuberculomas) were prospectively evaluated with contrast material-enhanced MR imaging, single-voxel proton MR spectroscopy (TE = 135 ms), and diffusion-weighted imaging (b = 0, 500, and 1000 s/mm(2)) before surgery. RESULTS: MR spectroscopy could differentiate benign from malignant tumors but was not useful in grading malignant tumors. In the differentiation of malignant from benign tumors, N-acetylaspartate (NAA)/choline (Cho), NAA/Cho + creatine (Cr), lactate/Cr, and alanin/Cr ratios (P <.001) were statistically more significant than NAA/Cr and lactate/lipid ratios (P <.05). Increase in lipid/Cr and alanin/Cr ratios could distinguish metastasis and meningiomas from other tumors, respectively (P <.001). DNETs could be diagnosed by their normal spectra and high ADC values (116.25 +/- 6.93 x 10(-3) mm(2)/s). Increase in lactate/Cr ratio correlated with degree of malignancy (r = -0.71). ADCs were effective for grading malignant tumors (P <.001) but not for distinguishing different tumor types with the same grade. High-grade malignant tumors (87.16 +/- 10.41 x 10(-3) mm(2)/s) had significantly lower ADC values than did low-grade malignant (115.33 +/- 11.67 x 10(-3) mm(2)/s) and benign (107.69 +/- 8.05 x 10(-3) mm(2)/s) tumors. Peritumoral ADCs were significantly higher in low-grade than in high-grade astrocytomas (P <.05). CONCLUSION: Combination of calculated ADC values from tumoral core and specific relative metabolite ratios acquired by MR spectroscopy added more information to MR imaging in the differentiation and grading of brain tumors and were more useful together than each alone. PMID- 12591639 TI - Concordant pre- and postsynaptic deficits of dopaminergic neurotransmission in neurologic Wilson disease. AB - BACKGROUND AND PURPOSE: Although previous brain imaging studies of Wilson disease (WD) focused on the dopaminergic system, correlational data on the integrity of the pre- and postsynaptic compartments are lacking. The present study was initiated to intra-individually determine the integrity of these compartments in patients with WD. METHODS: A total of 46 patients with WD and 10 matched control subjects underwent [(123)I]2beta-carbomethoxy-3beta-(4[(123)I]iodophenyl)tropane ([(123)I]beta-CIT) and [(123)I]iodobenzamide ([(123)I]IBZM) single photon emission CT (SPECT). For both radiotracers, specific striatal binding ratios (with the cerebellum as the reference region) were calculated after a standardized region-of-interest technique was applied. In addition, the severity of putative neurologic symptoms was evaluated by using a linear scoring system. RESULTS: In patients without neurologic symptoms, striatal binding ratios of both radiotracers did not differ from those of the control group (13.8 +/- 3.1 vs 12.0 +/- 3.4 and 2.00 +/- 0.19 vs 1.90 +/- 0.27; n.s.). In symptomatic patients, however, striatal binding ratios for both [(123)I]beta-CIT and [(123)I]IBZM were significantly reduced (9.1 +/- 2.3 and 1.64 +/- 0.18; P <.001). In all patients with WD, the [(123)I]beta-CIT and [(123)I]IBZM binding ratios were significantly correlated (r = 0.65, P <.001), as were SPECT parameters and the severity of the neurologic symptoms (r = -0.60 and -0.62; P <.001). CONCLUSION: These findings of a concordant bicompartmental dopaminergic deficit in neurologic WD provide in vivo evidence for assigning WD to the group of secondary Parkinsonian syndromes. These results could be relevant in therapeutic decision making in patients with this copper deposition disorder. PMID- 12591640 TI - Pattern of atherosclerotic carotid stenosis in Korean patients with stroke: different involvement of intracranial versus extracranial vessels. AB - BACKGROUND AND PURPOSE: The clinical pattern of stroke and the angiographic distribution of cerebral atherosclerosis in the Japanese and Chinese are different from those in whites. Our purpose was to evaluate the location and distribution of severe atherosclerotic stenoses in Korean patients by using cerebral angiography. METHODS: We retrospectively reviewed the cerebral angiographic findings in 268 patients (219 male, 49 female; mean age, 56 years) with one or more severe atherosclerotic stenoses (> or =70%), as shown on angiograms. These patients were selected from 1436 patients who were examined between 1996 and 1997. The analysis focused on the intracranial or extracranial location of the stenosis, the anterior and posterior circulations, and the multiplicity of the lesions. Statistical analysis was performed by using the chi(2) test. The data were then compared with data reported in other races and ethnic groups. RESULTS: A total of 389 severe stenotic sites were detected in 268 patients. A single stenosis was found in 56 (21%), and multiple stenoses were found in 212 (79%). Lesions were located in the intracranial area in 52% and in the extracranial area in 48%. They were detected in anterior circulation in 59% and in posterior circulation in 41%. Thirty-seven (66%) of 56 single stenosis were located in the intracranial area, and 19 (34%) were in the extracranial area. Of 333 lesions, 167 (50%) were multiple stenoses in the extracranial area, and 166 lesions (50%) were located in the intracranial vessels. The prevalence of intracranial stenosis was significantly higher in the single-stenosis group than in the multiple stenosis group (P <.05). CONCLUSION: Korean patients with severe atherosclerotic stenoses tend to have more intracranial stenoses. In particular, those with an isolated stenosis have more intracranial stenoses, compared with those with multiple stenoses. PMID- 12591641 TI - Conventional and perfusion MR imaging of parafalcine chondrosarcoma. AB - Intracranial chondrosarcomas have a predilection for the skull base, for which CT and MR imaging findings have been described. We present a rare case of primary chondrosarcoma arising from the falx in a young woman with no history of radiation. The CT, conventional MR imaging, perfusion MR imaging, and digital subtraction angiography findings are described. PMID- 12591642 TI - Marchiafava-Bignami disease: longitudinal MR imaging and MR spectroscopy study. AB - A case of Marchiafava-Bignami disease was serially evaluated with MR imaging and MR spectroscopy at 1, 2, 4, and 11 months after the onset of symptoms. The first MR imaging study showed extensive abnormal signal intensity of the corpus callosum without macroscopic changes; a diagnosis of Marchiafava-Bignami disease was made, and vitamin therapy was initiated. Follow-up studies showed progressive reduction of signal intensity abnormalities and residual callosal atrophy. MR spectroscopy revealed progressive reduction of the N-acetylaspartate:creatine ratio, with partial recovery in the last study, and a normalization of the choline:creatine ratio, which was initially slightly increased. Lactate was detectable during the subacute phase and was replaced by lipids after 4 months. This study confirmed the role of MR imaging in diagnosing Marchiafava-Bignami disease and particularly the value of MR spectroscopy in focusing the pathogenesis of the disease, monitoring its evolution and changes related to therapy. PMID- 12591643 TI - Pseudo-subarachnoid hemorrhage: a potential imaging pitfall associated with diffuse cerebral edema. AB - We report CT findings in seven patients with diffuse cerebral edema and increased attenuation in the basilar cisterns resembling subarachnoid hemorrhage. On the basis of autopsy (three cases) and lumbar puncture (four cases) findings, true subarachnoid hemorrhage was reasonably excluded. Pathophysiologic changes that occur with diffuse cerebral edema are explored, with proposed explanations for the appearance of a pseudo-subarachnoid hemorrhage. PMID- 12591644 TI - Coiling of very large or giant cerebral aneurysms: long-term clinical and serial angiographic results. AB - BACKGROUND AND PURPOSE: Initial complete occlusion of very large or giant aneurysms often cannot be accomplished, and most will partially reopen over time. This study was performed to assess the clinical and angiographic outcome of patients with very large or giant cerebral aneurysms treated with detachable coils. METHODS: During 6 years, 29 patients with 31 very large or giant (20-55 mm) cerebral aneurysms were initially treated with detachable coils. Nineteen patients presented with subarachnoid hemorrhage (SAH), and eight patients had symptoms of mass effect. One patient had an incidental aneurysm, and one patient had an additional aneurysm. RESULTS: Twenty-three (79%) of 29 patients had a good clinical outcome at a median follow-up of 50 months. One of 19 patients presenting with SAH had repeat bleed (annual rebleeding rate, 1.45%). After initial coiling, seven of 31 aneurysms were incompletely occluded; this rate increased to 20 of 29 aneurysms at 6-month follow-up angiography. After 16 repeat coiling procedures in 13 aneurysms, 12 of 29 aneurysms in surviving patients were still incompletely occluded. After additional treatment other than coiling (parent-vessel occlusion and/or surgery) in eight aneurysms, three of 25 aneurysms in 24 surviving patients were incompletely occluded. Only 13 (42%) of 31 aneurysms had one coiling as a sole therapy. CONCLUSION: Coiling of very large or giant aneurysms can be considered. Long-term clinical outcomes were good in 79% of patients. The stability of the coil mesh over time was poor, requiring repeat coiling, surgery, and/or parent-vessel balloon occlusion in 58% of the aneurysms primarily treated with coils. PMID- 12591645 TI - Broad-based intracranial aneurysms: thrombosis induced by stent placement. AB - For broad-based intracranial aneurysms, both surgical and endovascular treatments are demanding. Flexible metallic stents provide an alternative treatment option for these patients, because they enable coil placement through the stent mesh. Evidence from extracranial arteries indicates that aneurysms may also spontaneously thrombose after stent placement. This case report documents three patients with saccular intracranial aneurysms in whom aneurysm thrombosis occurred after stent placement only, without additional packing of the aneurysms with coils. PMID- 12591646 TI - Fluoroscopy fade for embolization of vein of Galen malformation. AB - A fluoroscopy fade technique in the embolization of a vein of Galen malformation in a 10-week-old infant is discussed herein. The availability of the fluoroscopy fade function eliminated the necessity of road mapping during the procedure and permitted minimal usage of contrast material and minimal radiation exposure for the patient. Techniques used during the embolization and the benefits of the fluoroscopy fade feature are also described. PMID- 12591647 TI - Simultaneous dual vessel cerebral angiography in gamma knife planning. AB - Gamma knife radiosurgery is an effective technique for treating arteriovenous malformations. However, treatment failures can occur as a result of incomplete visualization of the arteriovenous malformation nidus. We describe the use of bilateral femoral artery catheterization and simultaneous dual vessel cerebral angiography to facilitate treatment planning during gamma knife treatment. PMID- 12591648 TI - Stiff guide technique: technical report and illustrative case. AB - Endovascular techniques are gaining wider acceptance in the treatment of intracranial lesions. Tortuous vasculature is a common reason for failure to treat an intracranial lesion, especially when balloon catheters or stents are used. In these cases, the guiding catheter often buckles into the aorta during an attempt to place the balloon or stent. In our experience, a stiff wire can be used to support the guiding catheter, allowing the balloon catheter or stent to more readily navigate tortuous vessels. PMID- 12591649 TI - MR imaging features of nasopharyngeal tuberculosis: report of three cases and literature review. AB - The MR imaging appearances in three cases of nasopharyngeal tuberculosis are reported, and the findings are combined with three additional cases from a review of the literature. Two patterns of nasopharyngeal tuberculosis were identified. The first pattern is a discrete polypoid mass in the adenoids, and the second pattern is a more diffuse soft-tissue thickening of one or two of the walls of the nasopharynx. Extension outside the confines of the nasopharynx was not a feature, except in one case with early involvement of the prevertebral muscles. PMID- 12591650 TI - Giant infiltrating lipoma of the face: CT and MR imaging findings. AB - Infiltrating lipoma is an uncommon mesenchymal neoplasm that characteristically infiltrates adjacent tissues and tends to recur after excision. This type of lipoma is extremely rare in the head and neck region. We report a case of a giant infiltrating lipoma of the face, studied with CT and MR imaging. PMID- 12591651 TI - Neurocutaneous melanomatosis with a rapidly deteriorating course. AB - Neurocutaneous melanosis is a rare congenital syndrome characterized by large or multiple congenital melanocytic nevi and benign or malignant pigment cell tumors of the leptomeninges. The prognosis is extremely poor for symptomatic patients, even in the absence of malignant melanoma. We present serial MR imaging findings in the brain and spine of a child with congenital giant hairy nevi who developed progressive leptomeningeal melanomatosis and whose neurologic condition rapidly deteriorated. PMID- 12591653 TI - An allergic reaction to intraarterial nonionic contrast material. PMID- 12591654 TI - Imaging characteristics of bioterrorism: signs of our times. PMID- 12591655 TI - Radiologic manifestations of potential bioterrorist agents of infection. PMID- 12591657 TI - Hepatocellular carcinoma in cirrhotic livers: double-contrast thin-section MR imaging with pathologic correlation of explanted tissue. AB - OBJECTIVE: The aim of our study was to determine the sensitivity of double contrast MR imaging in the detection of hepatocellular carcinomas in patients with a cirrhotic liver. SUBJECTS AND METHODS: Thirty-one patients underwent double-contrast MR imaging and subsequent liver transplantation. Breath-hold T1- and T2-weighted MR images were obtained before and after administration of superparamagnetic iron oxide, and three-dimensional T1-weighted gradient-recalled echo MR images were obtained 10, 40, and 120 sec after a bolus injection of gadolinium. Hypervascular lesions that failed to take up superparamagnetic iron oxide were regarded as showing typical characteristics of hepatocellular carcinoma; lesions that had only one of these two characteristics (either hypervascularity or failure to take up superparamagnetic iron oxide) were regarded as highly suspicious for hepatocellular carcinoma. Radiology reports were correlated with pathology reports for the explanted livers. RESULTS: Thirty two hepatocellular carcinomas were found in 14 of the 31 patients. Combining the number of MR imaging reports citing lesions that were "typical of hepatocellular carcinoma" with the number of those citing lesions that were "highly suspicious," we found that for 25 of 32 lesions, an accurate MR imaging diagnosis of hepatocellular carcinoma was made (overall sensitivity, 78%). These lesions included 10 of the 11 lesions that were larger than 20 mm (sensitivity, 91%), 12 of the 13 lesions that were 11-20 mm (sensitivity, 92%), and three of the eight lesions that were 10 mm or less (sensitivity, 38%). Nineteen (76%) of 25 lesions had characteristics considered typical of hepatocellular carcinoma; the remaining six lesions either failed to take up superparamagnetic iron oxide and were hypovascular or were hypervascular but showed some uptake of superparamagnetic iron oxide. CONCLUSION: In patients with a cirrhotic liver, double-contrast MR imaging is highly sensitive in the diagnosis of hepatocellular carcinomas of 10 mm or larger, but success in the identification of tumors smaller than 10 mm is still limited. PMID- 12591658 TI - Physician satisfaction with high-resolution CT services provided by radiologists: results of a nationwide survey of pulmonary subspecialists. AB - OBJECTIVE: Our objectives were to measure the level of satisfaction of pulmonary medicine specialists who refer patients to radiology facilities for high resolution CT and to identify determinants of their reported satisfaction. MATERIALS AND METHODS: We surveyed 450 pulmonologist members of the American College of Chest Physicians. The self-administered questionnaire sought information about the radiologists and imaging facilities to which these clinicians referred patients for high-resolution CT of the lungs. The participants rated their satisfaction with the radiology services, estimated the number of patients referred for high-resolution CT per month, answered questions about certain attitudes and utilization practices, and provided general demographic information. RESULTS: Completed surveys were received from 230 pulmonologists practicing in 43 states. Satisfaction with high-resolution CT services was rated as follows: very satisfied (35% of respondents), satisfied (49%), and indifferent or dissatisfied (16%). A higher rating was found in pulmonologists in academic practice, in those who believed that the radiologists desired as much clinical information as possible, and in those who believed that two or more members of the radiology group were interested in high-resolution CT. Among satisfied pulmonologists, confidence in the radiologist's high-resolution CT interpretation was very important. Physician satisfaction was not significantly associated with the size of the radiology group or the number of monthly referrals for high-resolution CT. CONCLUSION: Pulmonologists in the United States appear to be satisfied with the high-resolution CT services provided by the radiologists in their communities. Satisfaction with radiology services might be further increased if radiologists expressed greater interest in high-resolution CT and pertinent clinical information and improved their interpretive skills. PMID- 12591659 TI - Medicare fraud and abuse. PMID- 12591661 TI - Whole-body three-dimensional multidetector CT of 13 Egyptian human mummies. AB - OBJECTIVE: The purpose of this article is to assess the role of multidetector CT and three-dimensional (3D) reconstructions in noninvasive studies of Egyptian mummies. MATERIALS AND METHODS: We studied 13 mummies from the Egyptian Museum in Torino, Italy, dating from Dynasty III to Dynasty IV (2650-2450 B.C.) and from the Ptolemaic period (332-30 B.C.) to the Roman period (30 B.C.-A.D. 395), using a multidetector CT unit with a single volumetric acquisition of the whole body, including lower extremities, followed by 3D reconstruction. All mummies were completely wrapped; preservation conditions of external wrappings were good in all. RESULTS: The general setting, embalming techniques, sex and age assessment (from body and skeletal features), anthropometric measurements (cranial measurements and evaluation of stature), conditions of the skeleton and soft tissue, any abnormalities, and the presence of foreign objects were evaluated in each mummy, and a detailed report was drawn up. Virtual unwrapping permitted the identification of physiognomy of the whole dehydrated body placed beneath the wrappings; 3D reconstruction and virtual fly-through navigation allowed further evaluations of the internal parts of the body. CONCLUSION: The results obtained with this protocol provided important anthropologic and paleopathologic information that would have been impossible to obtain by other noninvasive techniques. Moreover, this method has great potential for studies of conservation, anthropology, and paleopathology of other Egyptian and ancient human remains. Multidisciplinary cooperation among anthropologists, paleopathologists, Egyptologists, and radiologists is essential. PMID- 12591662 TI - Visualizing radiologic data. PMID- 12591663 TI - Imaging characteristics of cystic adventitial disease of the peripheral arteries: presentation as soft-tissue masses. AB - OBJECTIVE: Our purpose was to identify the characteristic imaging features of cystic adventitial disease of the peripheral arteries. CONCLUSION: Patients with cystic adventitial disease of the peripheral arteries often present for evaluation of soft-tissue masses involving the extremities. Noninvasive imaging reveals a characteristic appearance allowing confident diagnosis. PMID- 12591664 TI - Clinical evaluation and MR imaging features of popliteal artery entrapment and cystic adventitial disease. PMID- 12591665 TI - Association of biceps tendon tears with rotator cuff abnormalities: degree of correlation with tears of the anterior and superior portions of the rotator cuff. AB - OBJECTIVE: This study was performed to examine the relationship and association of abnormalities seen in the long head of the biceps brachii tendon to abnormal findings in the rotator cuff. MATERIALS AND METHODS: One hundred eleven patients underwent MR imaging for shoulder pain followed by arthroscopic or open shoulder surgery from January 1997 to December 2000. Patients were identified by a retrospective search, and all consecutive patients having undergone both MR imaging and surgery were included in the patient cohort. Official MR imaging interpretations were compared with operative reports, and all findings were recorded. RESULTS: Twenty-three patients were identified with partial- or full thickness tears of the long head of the biceps tendon. The sensitivity, specificity, and accuracy of unenhanced MR imaging of the shoulder for detecting these bicipital tears were 52%, 86%, and 79%, respectively. When a tear was present in the biceps tendon, the prevalence of supraspinatous, infraspinatus, and subscapularis tendon tears was 96.2%, 34.6%, and 47.1%, respectively. Patients with biceps tendon tears were significantly more likely to also have subscapularis tendon tears (p < 0.0001) and supraspinatous tendon tears (p < 0.008) than those patients who did not have biceps tendon tears. No significant relationship was found between the presence or absence of a biceps tendon tear and the presence or absence of a infraspinatus or teres minor tendon tear (p = 0.17). CONCLUSION: Tears of the long head of the biceps tendon have a statistically significant association with tears of the anterior and superior rotator cuff and are highly correlated with tears of the supraspinatous and subscapularis tendons. When tears of these tendons are detected, specific attention directed toward the long biceps tendon is warranted to characterize the status of this structure that provides additional stability to the shoulder joint. PMID- 12591666 TI - MR imaging features of osteochondritis dissecans of the femoral sulcus. AB - OBJECTIVE: The objective of this study was to describe the MR imaging features of osteochondritis dissecans of the femoral sulcus, which have not been described previously. CONCLUSION: Osteochondritis dissecans of the femoral sulcus has been recognized as a unique clinical and radiographic entity, warranting early diagnosis and appropriate treatment. Although this type of osteochondritis dissecans may be inconspicuous on radiography, fast spin-echo proton density weighted and T2-weighted MR imaging sequences allow evaluation for articular cartilage integrity and lesion stability. Because of the orientation of the femoral sulcus, osteochondritis dissecans involving the femoral sulcus is best evaluated on axial and sagittal images. PMID- 12591668 TI - MR imaging of the anatomy of and injuries to the lateral and posterolateral aspects of the knee. PMID- 12591669 TI - Posttraumatic cystlike defects of the scaphoid: late sign of occult microfracture and useful indicator of delayed union. AB - OBJECTIVE: To investigate the association between radiolucencies in the scaphoid and trauma, we studied the unenhanced radiographs and CT images of 1087 adult patients. CONCLUSION: Three (0.3%) of the 1087 patients had scaphoid radiolucencies that developed after trauma. We hypothesize that in these patients a microfracture of the scaphoid or subtle fractures are the cause of persisting clinical symptoms. Persistent loading of the scaphoid due to wrist movement can lead to unrestricted compressive forces and ischemia and to the development of a bone cyst-like pathologic change, apparent on the radiograph as a radiolucency and often misreported as a cyst. An alternate term, "cystlike defect" or "pseudocyst" of the scaphoid, is suggested. Such radiolucencies are indicators of previous microfractures or an untreated episode of trauma to the scaphoid. PMID- 12591670 TI - Imaging features of pseudoaneurysms of the hand in children and adults. AB - OBJECTIVE: Our objective was to review the imaging features of pseudoaneurysms of the hand in 25 patients. The patients presented with a mass, peripheral paresthesia, or ischemia. Pseudoaneurysm of the hand is a rare and often clinically unsuspected diagnosis. Correct diagnosis is important because there are risks for distal embolic disease with ischemia or gangrene of the fingers, ulnar or digital nerve dysfunction, rupture, or bone erosion and joint destruction. Scant reports appear in the world literature, and this report is the first review, to our knowledge, of the imaging features. The cause may be a history of a single direct trauma or chronic trauma, as seen in patients with hypothenar or thenar hammer syndrome. CONCLUSION: Awareness of the specific imaging appearances of pseudoaneurysms of the hand and their complications may improve the accuracy of radiologic diagnosis, advance the preoperative workup, and prevent possible clinical complications such as digital gangrene, nerve dysfunction, and aneurysm rupture. PMID- 12591671 TI - New radiographic classification of bone involvement in pedal mycetoma. AB - OBJECTIVE: The purpose of this study was to classify the pattern, extent, and severity of bone involvement in mycetoma of the foot. CONCLUSION: In this classification, stage 0 indicates the presence of soft-tissue swelling without bone involvement. Stage I refers to the extrinsic pressure effects on the intact bones in the vicinity of an expanding granuloma. Stage II results from irritation of the bone surface without actual intraosseous invasion. Cortical erosion and central cavitation occur in stage III. If the disease spreads longitudinally along a single ray, stage IV is established; horizontal spread along a single row represents stage V. Multidirectional spread due to uncontrolled infection is classified as stage VI. PMID- 12591672 TI - Focal nodular hyperplasia of the hematopoietic marrow simulating vertebral metastasis on FDG positron emission tomography. PMID- 12591673 TI - Effectiveness of MR imaging in characterizing small hepatic lesions: routine versus expert interpretation. AB - OBJECTIVE: The aim of our study was to compare the effectiveness of MR imaging characterization of small (1 cm), suboptimal debulking, and no response to chemotherapy (all Ps < 0.001). In univariate Cox survival analysis, high serum hK10 is associated with increased risk for relapse and death (hazard ratio = 2.59 and 3.15, respectively, P or =65 years of age hospitalized with ST-elevation myocardial infarction. We evaluated the distribution of patients in the 5 simple risk index groups, compared observed and published 30-day mortality rates, and assessed the score's discrimination and calibration. The simple risk index provided poor discrimination (c=0.62) and calibration (goodness of fit P<0.001) for survival at 30 days. Risk score distribution was skewed, because two thirds (66.1%) of all patients were classified in the highest-risk group, whereas fewer than 11.0% were classified in the 3 lowest-risk groups. Thirty-day mortality estimates were lower than those observed in the cohort (risk group 2 to 5: 1.9% to 17.4% versus 5.3% to 27.9%). Risk index discrimination, calibration, score distribution, and mortality estimates were worse among patients who did not receive acute reperfusion therapy than among those who did. CONCLUSIONS: The limited performance of the simple risk index highlights the limitations of applying prognostic models derived in RCT populations to the general population of patients 65 years and older. Prognostic scores must be validated in community-based cohorts before integration into clinical practice. PMID- 12591750 TI - Antianginal and antiischemic effects of ivabradine, an I(f) inhibitor, in stable angina: a randomized, double-blind, multicentered, placebo-controlled trial. AB - BACKGROUND: Heart rate reduction should benefit patients with chronic stable angina by improving myocardial perfusion and reducing myocardial oxygen demand. This study evaluated the antianginal and antiischemic effects of ivabradine, a new heart rate-lowering agent that acts specifically on the sinoatrial node. METHODS AND RESULTS: In a double-blind, placebo-controlled trial, 360 patients with a > or =3-month history of chronic stable angina were randomly assigned to receive ivabradine (2.5, 5, or 10 mg BID) or placebo for 2 weeks, followed by an open-label 2- or 3-month extension on ivabradine (10 mg BID) and a 1-week randomized withdrawal to ivabradine (10 mg BID) or placebo. Primary efficacy criteria were changes in time to 1-mm ST-segment depression and time to limiting angina during bicycle exercise (exercise tolerance tests), performed at trough of drug activity. In the per-protocol population (n=257), time to 1-mm ST-segment depression increased in the 5 and 10 mg BID groups (P<0.005); time to limiting angina increased in the 10 mg BID group (P<0.05). Deterioration in all exercise tolerance test parameters occurred in patients who received placebo during randomized withdrawal (all P<0.02) but not in those still receiving ivabradine. No rebound phenomena were observed on treatment cessation. CONCLUSIONS: Ivabradine produces dose-dependent improvements in exercise tolerance and time to development of ischemia during exercise. These results suggest that ivabradine, representing a novel class of antianginal drugs, is effective and safe during 3 months of use; longer-term safety requires additional assessment. PMID- 12591751 TI - Reductions in systemic and skeletal muscle blood flow and oxygen delivery limit maximal aerobic capacity in humans. AB - BACKGROUND: A classic, unresolved physiological question is whether central cardiorespiratory and/or local skeletal muscle circulatory factors limit maximal aerobic capacity (VO2max) in humans. Severe heat stress drastically reduces VO2max, but the mechanisms have never been studied. METHODS AND RESULTS: To determine the main contributing factor that limits VO2max with and without heat stress, we measured hemodynamics in 8 healthy males performing intense upright cycling exercise until exhaustion starting with either high or normal skin and core temperatures (+10 degrees C and +1 degrees C). Heat stress reduced VO2max, 2 legged VO2, and time to fatigue by 0.4+/-0.1 L/min (8%), 0.5+/-0.2 L/min (11%), and 2.2+/-0.4 minutes (28%), respectively (all P<0.05), despite heart rate and core temperature reaching similar peak values. However, before exhaustion in both heat stress and normal conditions, cardiac output, leg blood flow, mean arterial pressure, and systemic and leg O2 delivery declined significantly (all 5% to 11%, P<0.05), yet arterial O2 content and leg vascular conductance remained unchanged. Despite increasing leg O2 extraction, leg VO2 declined 5% to 6% before exhaustion in both heat stress and normal conditions, accompanied by enhanced muscle lactate accumulation and ATP and creatine phosphate hydrolysis. CONCLUSIONS: These results demonstrate that in trained humans, severe heat stress reduces VO2max by accelerating the declines in cardiac output and mean arterial pressure that lead to decrements in exercising muscle blood flow, O2 delivery, and O2 uptake. Furthermore, the impaired systemic and skeletal muscle aerobic capacity that precedes fatigue with or without heat stress is largely related to the failure of the heart to maintain cardiac output and O2 delivery to locomotive muscle. PMID- 12591752 TI - Myocardial proinflammatory cytokine expression and left ventricular remodeling in patients with chronic mitral regurgitation. AB - BACKGROUND: In an animal model, stretch was shown to induce myocardial tumor necrosis factor-alpha (TNF-alpha) expression. The purposes of this study were to determine whether the left ventricular (LV) volume overload that occurs in patients with chronic mitral regurgitation (MR) can induce myocardial and systemic TNF-alpha expression and whether there is a relationship between TNF alpha expression and LV remodeling. METHODS AND RESULTS: Plasma TNF-alpha and its receptors were measured before mitral valve (MV) repair surgery in 26 MR patients and 23+/-12 months after MV repair surgery in 9 MR patients. Myocardial mRNA copies of TNF-alpha were determined in 11 MR and 10 donor hearts using quantitative RT-PCR. Compared with 15 control subjects, pre-MV repair plasma TNF alpha (3.59+/-1.81 versus 2.03+/-1.02 pg/mL, P<0.005) and its receptor levels were elevated in MR patients. Myocardial TNF-alpha mRNA copies (corrected for beta-actin mRNA expression) in MR patients and donor hearts were 38.96+/ 42.74x10(6) and 0.88+/-0.75x10(6), respectively (P=0.01). After MV surgery, there was a decrease in the plasma levels of TNF-alpha (2.79+/-1.14 versus 3.51+/-1.34 pg/mL, P=0.02) and its receptors. There was a correlation between myocardial TNF alpha expression and preoperative LV end-diastolic and end-systolic volumes. Moreover, there was an inverse correlation between myocardial TNF-alpha expression and regression in LV end-diastolic (r=-0.76, P=0.007) and end-systolic (r=-0.73, P=0.01) volumes after MV surgery. CONCLUSIONS: TNF-alpha is expressed in the myocardium and plasma of MR patients. Correction of the LV volume overload with MV surgery results in reversal of TNF-alpha expression. There is a relationship between TNF-alpha expression and parameters of LV remodeling, suggesting that TNF-alpha may play a role in the pathogenesis of the LV remodeling that occurs in MR. PMID- 12591753 TI - Exercise stress test amplifies genotype-phenotype correlation in the LQT1 and LQT2 forms of the long-QT syndrome. AB - BACKGROUND: Experimental studies suggest that the interval between peak and end of T wave (Tpe) in transmural ECGs reflects transmural dispersion of repolarization (TDR), which is amplified by beta-adrenergic stimulation in the LQT1 model. In 82 patients with genetically identified long-QT syndrome (LQTS) and 33 control subjects, we examined T-wave morphology and various parameters for repolarization in 12-lead ECGs including corrected QT (QTc; QT/R-R(1/2)) and corrected Tpe (Tpec; Tpe/R-R(1/2)) before and during exercise stress tests. METHODS AND RESULTS: Under baseline conditions, LQT1 (n=51) showed 3 cardinal T wave patterns (broad-based, normal-appearing, late-onset) and LQT2 (n=31) 3 patterns (broad-based, bifid with a small or large notch). The QTc and Tpec were 510+/-68 ms and 143+/-53 ms in LQT1 and 520+/-61 ms and 195+/-69 ms in LQT2, respectively, which were both significantly larger than those in control subjects (402+/-36 ms and 99+/-36 ms). Both QTc and Tpec were significantly prolonged during exercise in LQT1 (599+/-54 ms and 215+/-46 ms) with morphological change into a broad-based T-wave pattern. In contrast, exercise produced a prominent notch on the descending limb of the T wave, with no significant changes in the QTc and Tpec (502+/-82 ms and 163+/-86 ms: n=19) in LQT2. CONCLUSIONS: Tpe interval increases during exercise in LQT1 but not in LQT2, which may partially account for the finding that fatal cardiac events in LQT1 are more often associated with exercise. PMID- 12591754 TI - Pulmonary vein diameter reduction after radiofrequency catheter ablation for paroxysmal atrial fibrillation evaluated by contrast-enhanced three-dimensional magnetic resonance imaging. AB - BACKGROUND: Radiofrequency catheter ablation (RFCA) is a promising intervention to treat atrial fibrillation. However, pulmonary vein (PV) stenosis after RFCA has been reported. The aim of this study was to investigate the incidence and time course of pulmonary vein stenosis after RFCA within a period of 3 months. Contrast-enhanced magnetic resonance angiography (MRA) was used to visualize pulmonary veins and was compared with radiographic angiography. METHODS AND RESULTS: Forty-six consecutive patients with symptomatic paroxysmal atrial fibrillation had RFCA in the orifice of 138 pulmonary veins. Comparison of diameters measured in 44 untreated vessels either by radiographic angiography or with MRA established the reliability of MRA (r=0.934). MRA measurements revealed an incidence of relevant diameter reductions of > or =25% or stenosis of > or =50% after RFCA of 25 of 138 (18.1%) treated vessels 1 day and/or 3 months after ablation. A progression of diameter reduction after RFCA was observed in 8.3% (maximum 75%), whereas a regression was observed in 6.3% of treated PVs. Ablation at a radial angle of >180 degrees of a pulmonary vein orifice increased the risk of diameter reduction significantly compared with ablation at a radial angle < or =180 degrees (P=0.002). CONCLUSIONS: The occurrence and progression of PV stenosis is a potential significant complication of RFCA in the orifice of pulmonary veins. These findings may have an impact on the technical performance of this intervention. In addition, long-term studies will be necessary to evaluate lumen reduction over time. MRA is a noninvasive, reproducible imaging modality for this purpose. PMID- 12591755 TI - Quantitative magnetic resonance imaging analysis of neovasculature volume in carotid atherosclerotic plaque. AB - BACKGROUND: Neovasculature within atherosclerotic plaques is believed to be associated with infiltration of inflammatory cells and plaque destabilization. The aim of the present investigation was to determine whether the amount of neovasculature present in advanced carotid plaques can be noninvasively measured by dynamic, contrast-enhanced MRI. METHODS AND RESULTS: A total of 20 consecutive patients scheduled for carotid endarterectomy were recruited to participate in an MRI study. Images were obtained at 15-second intervals, and a gadolinium contrast agent was injected coincident with the second of 10 images in the sequence. The resulting image intensity within the plaque was tracked over time, and a kinetic model was used to estimate the fractional blood volume. For validation, matched sections from subsequent endarterectomy were stained with ULEX and CD-31 antibody to highlight microvessels. Finally, all microvessels within the matched sections were identified, and their total area was computed as a fraction of the plaque area. Results were obtained from 16 participants, which showed fractional blood volumes ranging from 2% to 41%. These levels were significantly higher than the histological measurements of fractional vascular area. Nevertheless, the 2 measurements were highly correlated, with a correlation coefficient of 0.80 (P<0.001). CONCLUSIONS: Dynamic contrast-enhanced MRI provides an indication of the extent of neovasculature within carotid atherosclerotic plaque. MRI therefore provides a means for prospectively studying the link between neovasculature and plaque vulnerability. PMID- 12591756 TI - Immunosuppressive therapy for active lymphocytic myocarditis: virological and immunologic profile of responders versus nonresponders. AB - BACKGROUND: The beneficial effect of immunosuppressive treatment on myocarditis is still controversial, possibly because the immunologic and virological profile of potential candidates is largely unknown. METHODS AND RESULTS: Out of 652 biopsied patients, 112 had a histological diagnosis of active lymphocytic myocarditis; 41 of these 112 patients were characterized by progressive heart failure despite conventional therapy and were treated with prednisone and azathioprine for 6 months. All were resubmitted to cardiac catheterization, angiography, and endomyocardial biopsy at 1 and 6 months and followed-up for 1 year. A total of 21 patients responded with prompt improvement in left ventricular ejection fraction from 25.7+/-4.1% to 47.1+/-4.4% and showed evidence of healed myocarditis at control biopsy. Conversely, 20 patients failed to respond and showed a histological evolution toward dilated cardiomyopathy: 12 remained stationary, 3 underwent cardiac transplantation, and 5 died. We retrospectively performed a polymerase chain reaction on frozen endomyocardial tissue for the most common cardiotropic viruses and assessed circulating serum cardiac autoantibodies. Viral genomes were present in biopsy specimens of 17 nonresponders (85%), including enterovirus (n=5), Epstein-Barr virus (n=5) adenovirus (n=4), both adenovirus and enterovirus (n=1), influenza A virus (n=1), parvovirus-B19 (n=1), and in 3 responders, who were all positive for hepatitis C virus. Cardiac autoantibodies were present in 19 responders (90%) and in none of the nonresponders. CONCLUSIONS: In patients with active lymphocytic myocarditis, those with circulating cardiac autoantibodies and no viral genome in the myocardium are the most likely to benefit from immunosuppression. The beneficial effect of immunosuppression in hepatitis C virus myocarditis suggests a relevant immunomediated component of damage. PMID- 12591757 TI - Left ventricular mass and systolic performance in pediatric patients with chronic renal failure. AB - BACKGROUND: Children with chronic renal disease have a high prevalence of left ventricular hypertrophy (LVH), which is thought to be adaptive to improve contractility and lower wall stress in the face of increased afterload and preload. The aim of this study was to determine the association between LV mass, LV performance, and LV contractility in children with chronic renal insufficiency (CRI) and children undergoing chronic dialysis. METHODS AND RESULTS: Twenty-five children with CRI, 12 undergoing chronic dialysis, and 24 controls had echocardiographic evaluation during rest and peak exercise. LV performance was assessed by calculation of shortening fraction and heart rate-corrected velocity of circumferential fiber shortening (VCF). Contractility (VCF difference) was determined based on the relation between VCF and end-systolic wall stress. Contractile reserve was assessed by the difference between contractility at rest and peak exercise. The dialysis group had higher LVM index than the group with CRI (42.9+/-10.3 versus 29.9+/-9.4 g/m(2.7), P<0.001). Both groups had higher LVM index compared with controls (22.2+/-6.1 g/m(2.7), P<0.001). At rest, the CRI and dialysis groups had significantly higher VCF(c) (P<0.001) and VCF difference (P<0.05) and significantly lower wall stress (P<0.01) compared with the control group. Dialysis patients had significantly lower contractile reserve compared with the control group (P<0.03). CONCLUSIONS: These results indicate that children with CRI and undergoing chronic dialysis have increased LVM, LV performance, and contractility at rest. However, dialysis patients have diminished contractile reserve during exercise, which might be an indicator for the development of more severe systolic dysfunction over time. PMID- 12591758 TI - Failure of thrombus to resolve in urokinase-type plasminogen activator gene knockout mice: rescue by normal bone marrow-derived cells. AB - BACKGROUND: Monocytes may have an important role in the resolution of venous thrombosis. Increased expression of tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA) is associated with an ingress of monocytes into the thrombus. This study was designed to evaluate the importance of these activators in thrombus resolution. METHODS AND RESULTS: Inferior caval vein thrombosis was induced in cohorts of adult wild-type, uPA gene-knockout (uPA /-), and tPA gene-knockout (tPA-/-) mice in a flow model. Thrombi were harvested from wild-type and uPA-/- mice (n=60 per group) between 1 and 60 days. Thrombi were also obtained from groups of wild-type and tPA-/- mice (n=24 per group) between 1 and 28 days. Thrombus size and macrophage content were measured by computer-assisted image analysis. Thrombus resolution was significantly impaired in the uPA-/- mice compared with wild-type controls (P<0.0001) but was unaffected in tPA-/- mice. Monocyte content in wild-type mice was highest at 14 days after thrombus induction and was approximately 4 times greater than in uPA-/- mice (P=0.0043). Thrombus size in uPA-/- mice transplanted with wild-type marrow (0.29+/-0.06 mm2) was significantly smaller than in uPA-/- mice given uPA-/- bone marrow (3.9+/-1.1 mm2) (P=0.0022). Donor bone marrow-derived cells expressing LacZ were present in the thrombus after transplantation. CONCLUSIONS: The resolution of experimental venous thrombus is dependent on uPA but is unaffected by the absence of tPA. Absence of uPA is also associated with delayed monocyte recruitment into the thrombus. Transplanting wild-type bone marrow restores thrombus resolution in uPA-/- animals, suggesting an important role for bone marrow-derived cells in this process. PMID- 12591759 TI - Induction of coxsackievirus-adenovirus-receptor expression during myocardial tissue formation and remodeling: identification of a cell-to-cell contact dependent regulatory mechanism. AB - BACKGROUND: The coxsackievirus-adenovirus receptor (CAR) was cloned as a receptor for both viruses, but its primary biological functions and regulatory mechanisms are unknown. CAR was low in healthy adult myocardium, whereas strong CAR reexpression was observed in human dilated cardiomyopathy. The molecular mechanisms of CAR induction in cardiomyocytes are unknown. METHODS AND RESULTS: We report on CAR regulation during development, CAR induction after myocardial infarction, and cell-to-cell contact-dependent CAR regulation in the rat. The high CAR expression during development in various organs decreased up to 190-fold after birth. After infarction resulting in severe cardiac dysfunction (dP/dt(max), -53%; dP/dt(min), -58%; left ventricular pressure, -45%), CAR was induced locally in cardiomyocytes of the infarct zone, where it was also expressed by capillary-like CD31+ structures and CD18+ interstitial cells, whereas it remained confined to subendothelial layers of arterioles and venules. In cultured cardiomyocytes, endothelin-1, cardiotrophin-1, leukemia-inhibiting factor, and cyclic stretch had no effect on CAR, whereas at high versus low cell density, CAR was suppressed up to 10-fold (P=0.006). Conditioned media from low- or high-density cardiomyocytes or cardiofibroblasts had no effect. CONCLUSIONS: The locally confined CAR upregulation after infarction makes induction by various humoral factors unlikely, because cardiac dysfunction results in high activities of sympathetic and renin-angiotensin systems and cytokines. The cell culture experiments identify a cell-to-cell contact-dependent mechanism of CAR regulation. Further characterization of the signals linking cell-to-cell interactions to CAR gene expression may provide insight into mechanisms and functional consequences of the generalized CAR induction in dilated cardiomyopathy, and of its local induction after myocardial infarction. PMID- 12591760 TI - Defining the transmurality of a chronic myocardial infarction by ultrasonic strain-rate imaging: implications for identifying intramural viability: an experimental study. AB - BACKGROUND: In a correlative functional/histopathologic study, we investigated the regional deformation characteristics of both chronic nontransmural and transmural infarctions before and after a dobutamine challenge. METHODS AND RESULTS: After stenosing copper-coated stent implantation to produce circumflex artery endothelial proliferation, 18 pigs were followed up for 5 weeks. Posteuthanasia histology showed 10 to have a nontransmural and 8 a transmural infarction. Eight nonstented animals served as controls. Regional radial function was monitored by measuring ultrasound-derived peak systolic strain rates (SR(SYS)) and systolic strains (epsilon(SYS)) (1) before stent implantation and (2) at 5 weeks, at baseline (bs) and during an incremental dobutamine infusion. In controls, dobutamine induced a linear increase in SR(SYS) (dobutamine: bs, 4.8+/-0.4 s(-1); 20 microg x kg(-1) x min(-1), 9.9+/-0.7 s(-1); P<0.0001) and an initial increase of epsilon(SYS) at low dose (bs, 58+/-5%; at 5 microg x kg(-1) x min(-1), 78+/-6%; P<0.05) but a subsequent decrease during higher infusion rates. In the nontransmural group, bs SR(SYS) and epsilon(SYS) were significantly lower than prestent values (SR(SYS), 2.9+/-0.5 s(-1) and epsilon(SYS), 32+/-6%, P<0.05 versus prestent). During dobutamine infusion, SR(SYS) increased slightly at 5 microg x kg(-1) x min(-1) (4.7+/-0.6 s(-1), P<0.05) but fell during higher infusion rates, whereas epsilon(SYS) showed no change. For nontransmural infarctions, transmural scar extension correlated closely with epsilon(SYS) at bs (r=0.88). For transmural infarctions, SR(SYS) at bs was significantly reduced and epsilon(SYS) was almost not measurable (SR(SYS), 1.8+/-0.3 s(-1); epsilon(SYS), 3+/-4%). Both deformation parameters showed no further change during the incremental dobutamine infusion. CONCLUSIONS: Ultrasonic deformation values could clearly differentiate chronic nontransmural from transmural myocardial infarction. The transmural extension of the scar could be defined by the regional deformation response. PMID- 12591761 TI - Spironolactone and its main metabolite, canrenoic acid, block human ether-a-go-go related gene channels. AB - BACKGROUND: It has been demonstrated that spironolactone (SP) decreases the QT dispersion in chronic heart failure. In this study, the effects of SP and its metabolite, canrenoic acid (CA), on human ether-a-go-go-related gene (HERG) currents were analyzed. METHODS AND RESULTS: HERG currents elicited in stably transfected Chinese hamster ovary cells were measured with the whole-cell patch clamp technique. SP decreased HERG currents in a concentration-dependent manner (IC50=23.0+/-1.5 micromol/L) and shifted the midpoint of the activation curve to more negative potentials (Vh=-13.1+/-3.4 versus -18.9+/-3.6 mV, P<0.05) without modifying the activation and deactivation kinetics. SP-induced block (1 micromol/L) appeared at the range of membrane potentials coinciding with that of channel activation, and thereafter, it remained constant, reaching 24.7+/-3.8% at +60 mV (n=6, P<0.05). CA (0.01 nmol/L to 500 micromol/L) blocked HERG channels in a voltage- and frequency-independent manner. CA at 1 nmol/L shifted the midpoint of the activation curve to -19.9+/-1.8 mV and accelerated the time course of channel activation (tau=1064+/-125 versus 820+/-93 ms, n=11, P<0.01). The envelope of the tail test demonstrated that at the very beginning of the pulses to +40 mV (25 ms), a certain amount of block was apparent (31.3+/-9.9%). CA did not modify the voltage-dependence of HERG channel inactivation (Vh=-60.8+/-5.6 versus -62.9+/-3.1 mV, n=6, P>0.05) or the kinetics of the reactivation process at any potential tested. CA and aldosterone also blocked the native I(Kr) in guinea-pig ventricular myocytes. CONCLUSIONS: At concentrations reached after administration of therapeutic doses of SP, CA blocked the HERG channels by binding to both the closed and open states of the channel. PMID- 12591762 TI - Potent metalloporphyrin peroxynitrite decomposition catalyst protects against the development of doxorubicin-induced cardiac dysfunction. AB - BACKGROUND: Increased oxidative stress and dysregulation of nitric oxide have been implicated in the cardiotoxicity of doxorubicin (DOX), a commonly used antitumor agent. Peroxynitrite is a reactive oxidant produced from nitric oxide and superoxide in various forms of cardiac injury. Using a novel metalloporphyrinic peroxynitrite decomposition catalyst, FP15, and nitric oxide synthase inhibitors or knockout mice, we now delineate the pathogenetic role of peroxynitrite in rodent models of DOX-induced cardiac dysfunction. METHODS AND RESULTS: Mice received a single injection of DOX (25 mg/kg IP). Five days after DOX administration, left ventricular performance was significantly depressed, and high mortality was noted. Treatment with FP15 and an inducible nitric oxide synthase inhibitor, aminoguanidine, reduced DOX-induced mortality and improved cardiac function. Genetic deletion of the inducible nitric oxide synthase gene was also accompanied by better preservation of cardiac performance. In contrast, inhibition of the endothelial isoform of nitric oxide synthase with N-nitro-L arginine methyl ester increased DOX-induced mortality. FP15 reduced the DOX induced increase in serum LDH and creatine kinase activities. Furthermore, FP15 prevented the DOX-induced increase in lipid peroxidation, nitrotyrosine formation, and metalloproteinase activation in the heart but not NAD(P)H-driven superoxide generation. Peroxynitrite neutralization did not interfere with the antitumor effect of DOX. FP15 also decreased ischemic injury in rats and improved cardiac function and survival of mice in a chronic model of DOX-induced cardiotoxicity. CONCLUSIONS: Thus, peroxynitrite plays a key role in the pathogenesis of DOX-induced cardiac failure. Targeting peroxynitrite formation may represent a new cardioprotective strategy after DOX exposure or in other conditions associated with peroxynitrite formation, including myocardial ischemia/reperfusion injury. PMID- 12591763 TI - Widening of the excitable gap and enlargement of the core of reentry during atrial fibrillation with a pure sodium channel blocker in canine atria. AB - BACKGROUND: This study aimed to assess the effects of pilsicainide, a pure sodium channel blocker, on electrophysiological action and wavefront dynamics during atrial fibrillation (AF). METHODS AND RESULTS: In a newly developed model of isolated, perfused, and superfused canine atria (n=12), the right and left endocardia were mapped simultaneously by use of a computerized mapping system. AF was induced with 1 to 5 micromol/L acetylcholine. The antifibrillatory actions of pilsicainide on AF cycle length (AFCL), refractory period (RP), conduction velocity (CV), excitable gap (EG), and the core of the mother rotor were studied. The RP was defined as the shortest coupling interval that could capture the fibrillating atrium. The EG was estimated as the difference between the AFCL and RP. At baseline, multiple wavefronts were observed. After 2.5 microg/mL infusion of pilsicainide, all preparations showed irregular activity, and AF was terminated in 2 preparations. The AFCL and RP were prolonged, and CV was decreased significantly. The EG was widened (147%; P<0.01), and the core perimeter was increased (100%; P<0.01). Increasing the dosage either terminated AF (6 preparations) or converted to organized activity (ie, atypical atrial flutter) (4 preparations). On the maps, all "unorganized" AFs were terminated with the excitation of the core of the mother rotor by an outside wavefront, whereas in preparations with atrial flutter, pilsicainide did not terminate its activity. CONCLUSIONS: Widening of the EG by pilsicainide facilitates the excitation of the core of the mother rotor, leading to the termination of AF. In some experiments, pilsicainide converts AF to persistent atrial flutter. PMID- 12591765 TI - Radiation dose in computed tomography of the heart. PMID- 12591764 TI - Rapamycin induces heme oxygenase-1 in human pulmonary vascular cells: implications in the antiproliferative response to rapamycin. AB - BACKGROUND: Rapamycin is an immunosuppressive agent with antiproliferative properties against not only lymphocytes but also vascular endothelial and smooth muscle cells, and it reduces the fibroproliferative response to vascular injury. Heme oxygenase-1 (HO-1) has also been shown to have graft protective effects and to inhibit vascular remodeling. In this study, we evaluated whether there is an interaction between rapamycin and HO-1. METHODS AND RESULTS: In human pulmonary artery endothelial or smooth muscle cells, HO-1 expression was evaluated in response to rapamycin or wortmannin, an inhibitor of the upstream modulator of mammalian target of rapamycin (mTOR) PI-3K. We also evaluated whether the inhibitory actions of rapamycin on platelet-derived growth factor-dependent proliferation was mediated by HO using the chemical inhibitor tin protoporphyrin. Rapamycin induced HO-1 expression in both pulmonary endothelial and smooth muscle cells, whereas no to little increase was seen in response to another immunosuppressive agent, cyclosporin A. HO-1 expression was also increased in response to wortmannin, suggesting that the PI-3K-mTOR pathway is required for this induction. Inhibition of HO activity resulted in a loss of the antiproliferative activity of rapamycin in growth factor-stimulated smooth muscle cells. CONCLUSIONS: The induction of HO-1 expression by rapamycin and, more importantly, the effects of tin protoporphyrin, an inhibitor of HO activity, on the antiproliferative actions of rapamycin suggest that the effects of rapamycin may be, at least in part, modulated by its actions on HO-1. PMID- 12591766 TI - Images in cardiovascular medicine. Generalized large vessel arteritis visualized by 18fluorodeoxyglucose-positron emission tomography. PMID- 12591767 TI - Cardiology patient page. Aortic aneurysm. PMID- 12591768 TI - Images in cardiovascular medicine. Coronary hematoma visualized by intravascular ultrasound and magnetic resonance imaging. PMID- 12591769 TI - Immunosuppression in myocarditis. PMID- 12591770 TI - Reasonable control: gun registration in Canada. PMID- 12591773 TI - Evaluating elective surgery. PMID- 12591774 TI - Evaluating elective surgery. PMID- 12591776 TI - Evaluating elective surgery. PMID- 12591778 TI - Neuroimaging misinformation. PMID- 12591779 TI - An observational study comparing 2-hour 75-g oral glucose tolerance with fasting plasma glucose in pregnant women: both poorly predictive of birth weight. AB - BACKGROUND: The definition and treatment of glucose intolerance during pregnancy are matters of intense controversy. Our goal was to examine the value of the 75-g oral glucose tolerance test (OGTT) in terms of its ability to predict birth weight percentile in a group of women with singleton pregnancies who received minimal treatment for their glucose intolerance. METHODS: We reviewed the results of OGTTs performed between 24 and 28 weeks' gestation in a group of 300 consecutive high-risk women (mean age 29.5 years [95% confidence interval, CI, 28.9-30.1]; parity 1.5 [95% CI 1.4-1.7]) whose plasma glucose level 1 hour after a randomly administered 50-g glucose load was 8.0 mmol/L or above. These data were compared with results for a randomly selected control group of 300 women whose plasma glucose level 1 hour after a 50-g glucose load was less than 8.0 mmol/L (mean age 28.0 years [95% CI 27.4-28.6]; parity 1.5 [95% CI 1.3-1.6]). RESULTS: For 76 (25.3%) of the 300 high-risk women, the plasma glucose level 2 hours after a 75-g glucose load (confirmatory OGTT) was 7.8 mmol/L or more, but only 6 of these were treated with insulin, which emphasizes the low level of intervention in this group. Thirty (10.0%) of the neonates in this group were large for gestational age (LGA; adjusted weight at or above the 90th percentile). This proportion did not significantly differ from the proportion for the control group (25 or 8.3%). After exclusion of the 6 insulin-treated women, simple correlations between birth weight percentile and fasting or 2-hour plasma glucose levels were very weak (r = 0.23 and 0.16 respectively; p < 0.01). The correlation between birth weight percentile and fasting or 2-hour plasma glucose persisted in a multiple regression analysis that included the following maternal variables: age, prepregnancy weight, weight gain during pregnancy, parity and smoking. In the multivariate models, the standardized coefficients for fasting and 2-hour plasma glucose levels were low (r = 0.19 [p < 0.001] and r = 0.13 [p = 0.02] respectively). These multivariate models could not explain more than 22% of the total variability in birth weight percentile. INTERPRETATION: In this population of pregnant, untreated diabetic women, plasma glucose levels (either fasting or after various glucose loads) were independently but poorly correlated with birth weight; no more than 3% to 5% of birth weight variability could be explained by changes in glucose tolerance. Fasting plasma glucose was consistently but marginally better than the plasma glucose level 2 hours after 75-g glucose load for predicting LGA neonates. We conclude that neonatal macrosomia is influenced by variables that are largely independent of plasma glucose concentrations. PMID- 12591780 TI - Equity in Canadian health care: does socioeconomic status affect waiting times for elective surgery? AB - BACKGROUND: Waiting times for surgical and other procedures are an important measure of how well the health care system responds to patient needs. In a universal health care system such as Canada's, it is important to determine if waiting times vary by socioeconomic status (SES). We compared waiting times for elective surgery of patients living in low and high socioeconomic areas. METHODS: We reviewed the medical charts of all patients who underwent elective surgery at a Canadian academic health centre between 1992 and 1999. Using patient postal codes we assigned SES on the basis of 5 characteristics in the 1996 census data. We compared waiting times for surgery for people from regions in the lowest third (low SES group) with that for patients from regions in the upper third (high SES group). RESULTS: On average, patients in the high SES group waited 31.1 days and those in the low SES group waited 29.3 days. When differences in waiting times for 22 common procedures were examined between the groups, only the difference for prostatectomy was statistically significant: patients in the high SES group waited 4.4 fewer days than those in the low SES group. INTERPRETATION: We found little evidence that residing in a region in which SES was in the lowest third was associated with longer waiting times for elective surgery. PMID- 12591781 TI - What happens to patients who leave hospital against medical advice? AB - BACKGROUND: Patients who leave hospital against medical advice (AMA) may be at risk of adverse health outcomes and readmission. In this study we examined rates of readmission and predictors of readmission among patients leaving hospital AMA. METHODS: We prospectively studied 97 consecutive patients who left the general medicine service of an urban teaching hospital AMA. Each patient was matched according to age, sex and primary diagnosis with a control patient who was discharged routinely. Readmission rates were examined using Kaplan-Meier analysis. Regression models were used to test the hypothesis that readmissions among patients discharged AMA followed a biphasic curve. RESULTS: Patients who left AMA were much more likely than the control patients to be readmitted within 15 days (21% v. 3%, p < 0.001). Readmissions occurred at an accelerated pace during the first 15 days, followed by a 75-day period during which readmissions occurred at a rate comparable to that among the control patients. Among the patients who left AMA, being male and having a history of alcohol abuse were significant predictors of readmission within 15 days; however, these characteristics were common among the patients who left AMA. In the Cox proportional hazard models, leaving AMA was the only significant predictor of readmission (adjusted hazard ratio 2.5, 95% confidence interval 1.4-4.4). INTERPRETATION: The significantly increased risk of readmission among general medicine patients who leave hospital AMA is concentrated in the first 2 weeks after discharge. However, it is difficult to identify which patients will likely be readmitted. PMID- 12591782 TI - Incidence, clearance and predictors of human papillomavirus infection in women. AB - BACKGROUND: Persistent infection with carcinogenic human papillomavirus (HPV) is linked to high-grade lesions and cervical cancer. To better understand the natural history of HPV, we sought to determine the rates of incident and cleared carcinogenic HPV infection, by age, among women aged 15-49 years and to explore risk factors for incident infection. METHODS: Women enrolled in an earlier HPV prevalence survey (500 of 800 who were HPV-negative and all 121 who were HPV positive) were invited to participate in follow-up HPV testing at their periodic health examination one year later. A cervical soft-brush specimen for HPV testing and a smear for cytologic examination were obtained, and participants completed a questionnaire on their demographic characteristics and sexual history. RESULTS: Two hundred and fifty-three (50.6%) previously HPV-negative women and 54 (44.6%) previously HPV-positive women were retested. The mean interval between visits was 14.0 (standard deviation 2.0, median 13.5, range 9.0-21.3) months. Incident HPV infection occurred in 11.1% (28/253) of the women overall, with the highest rate, 25.0% (6/24), in the 15-19-year age group. In the univariate analyses, risk factors for incident HPV were the median number of sexual partners in the past year (< or = 1 v. > or = 2: odds ratio [OR] 8.2, 95% confidence interval [CI] 3.0 22.2; p < 0.001) and the median number of sexual partners over a lifetime (> 3 v. < or = 3: OR 3.0, 95% CI 1.2-7.2; p = 0.014). In multivariate logistic regression modelling adjusted for age, median number of sexual partners in the past year, median number of sexual partners over a lifetime, marital status, current smoking and current use of oral contraceptives, only the median number of sexual partners in the past year remained significantly associated with incidence (OR 6.2, 95% CI 1.6-24.5; p = 0.009). Of the previously HPV-positive women, 51.9% (28/54) had cleared the infection. INTERPRETATION: Incident infection with carcinogenic HPV was highest in women aged 15-19 years, and risk factors were consistent with a sexually transmitted infection. A large proportion of the women who were HPV positive appeared to have cleared the infection after one year. PMID- 12591783 TI - Does screening for gestational diabetes mellitus make a difference? PMID- 12591784 TI - Cost-utility analysis: use QALYs only with great caution. PMID- 12591785 TI - Physicians and nuclear disarmament. PMID- 12591786 TI - Clinical practice guidelines for the care and treatment of breast cancer: breast radiotherapy after breast-conserving surgery (summary of the 2003 update). PMID- 12591787 TI - Diagnosis and management of epilepsy. AB - This article concisely describes the more common epilepsy conditions and will enable physicians to efficiently evaluate and manage these disorders. Salient aspects of the history and examination, together with electroencephalography, will usually determine the epilepsy syndrome (category), forming the basis for any further investigation and possible antiepileptic therapy. Imaging may be required in some circumstances. PMID- 12591789 TI - Pertussis in adults. PMID- 12591788 TI - A cryptic cause of cryptococcal meningitis. AB - Cryptococcus neoformans commonly causes opportunistic infection in immunocompromised patients, especially in patients with AIDS. The CD4+ T lymphocyte count is measured in patients with HIV infection, because it signals an increased risk of opportunistic infection and a decline in immunological function. We report a case of cryptococcal meningitis in a patient with persistently low CD4+ cell counts without evidence of HIV infection. The patient's underlying immunocompromised state was attributed to idiopathic CD4+ T lymphocytopenia (ICL), a recently described syndrome characterized by depletions in the CD4+ T-cell subsets without evidence of HIV infection. Immunodeficiency can exist in the absence of laboratory evidence of HIV infection, highlighting the importance of evaluating T-cell subsets in patients who present with unusual infections. PMID- 12591790 TI - Diane-35 (cyproterone acetate): safety concerns. PMID- 12591791 TI - Looking at the living human growth plate. PMID- 12591797 TI - MD's affidavit decries company's use of prescription data mining. PMID- 12591799 TI - MD group criticizes possible US attack on Iraq. PMID- 12591800 TI - Saskatchewan first with mandatory reporting of medical errors. PMID- 12591802 TI - Malpractice fee hikes mean some US MDs can't afford to work. PMID- 12591804 TI - Are spouses the key to retention of rural MDs? PMID- 12591807 TI - CMA to revise Code of Ethics. PMID- 12591808 TI - Alberta steps up recruitment of foreign professionals. PMID- 12591809 TI - MDs' office Internet use hits 57%. PMID- 12591812 TI - Diagnosis of culture negative endocarditis: novel strategies to prove the suspect guilty. PMID- 12591813 TI - Prolonged asymptomatic sinus pause indicated by implantable loop recording. PMID- 12591814 TI - An incidental case of thrombus formation in a patient with a portacath inserted for regular blood transfusions. PMID- 12591815 TI - Management of prosthetic valve endocarditis: a clinical challenge. PMID- 12591816 TI - Misplaced pacemaker wire as a cause of mitral regurgitation. PMID- 12591817 TI - Outcome following percutaneous coronary intervention: not, so far, in our genes. PMID- 12591818 TI - The primacy of clinical effectiveness for cost effectiveness analysis. PMID- 12591819 TI - Erectile dysfunction and the cardiovascular patient: endothelial dysfunction is the common denominator. AB - Erectile dysfunction (ED) is a common condition and studies predict that it will become even more common in the future. There is increasing evidence to suggest that it is predominantly a vascular disease and may even be a marker for occult cardiovascular disease. The common pathological process is at the level of the endothelium, and cardiovascular risk factor control may be the key to preventing ED. Many men with established cardiovascular disease have ED. Specific guidelines for the management of ED in these patients have been produced by an expert panel. Cardiovascular risk stratification is an important initial step in managing such patients. In cardiac patients considered to have low cardiovascular risk, the management of ED can be safe and effective. PMID- 12591821 TI - A "natural experiment" in cardiovascular epidemiology in the early 21st century. AB - Despite similar traditional risk factors, morbidity and mortality rates from coronary heart disease in western and non-western cohorts remain substantially different. Careful study of such cohorts may help identify novel risk factors for CHD, and contribute to the formulation of new preventive strategies PMID- 12591822 TI - Multiple cystic aneurysms in aortitis demonstrated by three dimensional volume rendering images of multislice computed tomography. PMID- 12591823 TI - Blood culture negative endocarditis: analysis of 63 cases presenting over 25 years. AB - OBJECTIVE: To analyse cases of blood culture negative endocarditis (BCNE) seen at St Thomas' Hospital, London, between 1975 and 2000. METHODS: Data on all episodes of endocarditis with negative blood cultures seen at St Thomas' Hospital between 1975 and 2000 were collected prospectively and analysed retrospectively. RESULTS: Sixty three patients with BCNE were seen during the study period: 48 (76%) with native and 15 (24%) prosthetic valve infection. BCNE accounted for 12.2% of the 516 cases of endocarditis seen at St Thomas' Hospital. The diagnosis of endocarditis was clinically definite by the Duke criteria in only 21% (7 of 34) of cases of pathologically proven native valve endocarditis but in 62% (21 of 34) of cases by the St Thomas' modifications of the criteria. Comparable figures for the 11 cases of pathologically proven prosthetic valve endocarditis were 45% and 73%. Despite negative blood cultures a causative organism was identified in 31 (49%) of the 63 cases: in 15 by serology (8 Coxiella burnetii, 6 Bartonella species, and 1 Chlamydia psittaci); in 9 cases by culture of the excised valve; in 3 by microscopy of the excised valve, on which large numbers of Gram positive cocci were seen although the culture was sterile; and in the other 4 by isolation from a site other than the excised valve (2 respiratory specimens, 1 from the pacemaker tip, and 1 from an excised embolus). In addition 5 of the 6 cases of Bartonella infection were confirmed by polymerase chain reaction study of the excised valve. Two thirds of the 32 patients for whom no pathogen was identified had received antibiotics before blood was cultured. Thus truly "negative" endocarditis was very uncommon (6% of the cases). CONCLUSION: If blood cultures are negative in definite or suspected endocarditis, serum should be analysed for Bartonella, Coxiella, and Chlamydia species antibodies, and the excised valve or (rarely) embolus should be analysed by microscopy, culture, histology, and relevant polymerase chain reaction. Other specimens may be relevant. The Duke criteria performed poorly in BCNE; St Thomas' additional minor criteria gave more definite diagnoses. PMID- 12591824 TI - Left atrial thrombus caused by the primary antiphospholipid syndrome causing critical functional mitral stenosis. PMID- 12591826 TI - An unusual "right atrial mass" on echocardiography. PMID- 12591825 TI - Molecular diagnosis of culture negative infective endocarditis: clinical validation in a group of surgically treated patients. AB - OBJECTIVE: To assess the clinical validity of polymerase chain reaction (PCR) based molecular methods in the microbiological diagnosis of culture negative infective endocarditis in a group of surgically treated patients. DESIGN: Retrospective case-control study. SETTING: Reference cardiovascular surgical centre. PATIENTS AND SAMPLES: 15 culture negative patients with infective endocarditis classified according to Duke criteria, with 17 heart valve samples; 13 age and sex matched control patients without infective endocarditis, with 13 valve samples. INTERVENTIONS: Medical records were reviewed and clinical, demographic, and microbiological data collected, including results of molecular detection of bacteria and fungi from valve samples. The clinical validity of molecular diagnosis was assessed, along with the sensitivity and speed of the systems. RESULTS: In the study group, 14 patients were PCR positive (93%). Organisms detected were streptococci (3), staphylococci (2), enterobacter (1), Tropheryma whippelii (1), Borrelia burgdorferi (1), Candida albicans (1), and Aspergillus species (2). Three cases were positive on universal bacterial detection but the pathogen could not be identified because of contaminating background. One case was negative. All but two positive cases showed clinical correlations. These two cases had no symptoms of infective endocarditis but there was agreement with the surgical findings. All control cases were PCR negative. Results were available within eight hours, and if sequencing was necessary, within 48 hours. CONCLUSIONS: PCR based molecular detection of pathogens in valve samples from surgically treated culture negative infective endocarditis patients is fast, sensitive, and reliable. The technology, combined with thorough validation and clinical interpretation, may be a promising tool for routine testing of infective endocarditis. PMID- 12591827 TI - Prosthetic valve endocarditis: early and late outcome following medical or surgical treatment. AB - OBJECTIVE: To compare the early and late outcome of medical and surgical treatment in patients with prosthetic valve endocarditis within a single unit. DESIGN: All patients with proven prosthetic valve endocarditis treated in one institution between 1989 and 1999 were studied. RESULTS: There were 66 patients (24 female, 42 male), mean (SD) age 57 (14) years. Of these, 28 were treated with antibiotics alone and 38 with a combination of antibiotics and surgery. The in hospital mortality for the antibiotic group was 46% and for the surgical group, 24%. However, seven patients in the antibiotic group were considered too sick for curative treatment. The mortality in the remaining 21 medically treated patients (6/21; 29%) was not significantly different from that in the surgically treated patients (p = 0.15). Six patients in the medically treated group and one in the surgically treated group required late reoperation. Endocarditis recurred in three patients in the medically treated group, two of whom were treated surgically, and in one patient in the surgically treated group. Kaplan-Meier survival at 10 years was 28% in the medically treated group v 58% in the surgically treated group (p = 0.04). Freedom from endocarditis at five years was 60% in the surgically treated group and 65% in the medically treated group. CONCLUSIONS: Prosthetic valve endocarditis is a serious condition with high early and late mortality, irrespective of the treatment employed. These data show that selected patients with prosthetic valve endocarditis can be successfully treated with antibiotics alone. If required, surgery in this difficult group of patients can provide satisfactory freedom from recurrent infection. PMID- 12591828 TI - Effect of temperature on the Brugada ECG. PMID- 12591829 TI - Clinical criteria and the appropriate use of transthoracic echocardiography for the exclusion of infective endocarditis. AB - BACKGROUND: Clinical guidelines currently suggest that transthoracic echocardiography (TTE) be carried out in all patients with suspected endocarditis, but the use of TTE where there is a low probability of infective endocarditis has a poor diagnostic yield. This screening approach may no longer be appropriate. OBJECTIVE: To examine whether clinical criteria might aid decision making with respect to the use of TTE in possible endocarditis. DESIGN: A retrospective review of patient records. SETTING: Cardiology department of a tertiary referral centre. PATIENTS: 500 consecutive hospital inpatients referred for TTE to exclude endocarditis. MAIN OUTCOME MEASURES: Evidence of endocardial vegetations on TTE and the presence of predetermined clinical criteria that may predispose to, or be suggestive of, endocarditis. RESULTS: Evidence of infective endocarditis was detected on echocardiography in 43 of the 500 patients (8.6%). In 239 patients (48%), vegetations and certain prespecified clinical criteria were both absent. These criteria were: vasculitic/embolic phenomena; the presence of central venous access; a recent history of injected drug use; presence of a prosthetic valve; and positive blood cultures. The collective absence of these five criteria indicated a zero probability of TTE showing evidence of endocarditis. CONCLUSIONS: The use of simple clinical criteria during the decision making process may avoid many unnecessary TTE examinations in hospital inpatients with a low probability of endocarditis. PMID- 12591830 TI - Bangladeshi patients present with non-classic features of acute myocardial infarction and are treated less aggressively in east London, UK. AB - OBJECTIVE: To analyse differences in the presentation and management of Bangladeshi and white patients with Q wave acute myocardial infarction (AMI). DESIGN: Prospective observational study. SETTING: East London teaching hospital. PARTICIPANTS: 263 white and 108 Bangladeshi patients admitted with Q wave AMI. MAIN OUTCOME MEASURE: Character of presenting symptoms, their interpretation by the patient, and the provision of emergency treatment. RESULTS: There were no significant differences between Bangladeshi and white patients in the time from pain onset to hospital arrival (arrival time 64.5 (117.5) minutes v 63.0 (140.3) minutes, p = 0.63), but once in hospital it took almost twice as long for Bangladeshi as for white patients to receive thrombolysis (median (interquartile range) door to needle time 42.5 (78.0) minutes v 26.0 (47.7) minutes, p = 0.012). Bangladeshis were significantly less likely than whites to complain of central chest pain (odds ratio (OR) 0.11, 95% confidence interval (CI) 0.03 to 0.38; p = 0.0006) or to offer classic descriptions of the character of the pain (OR 0.25, 95% CI 0.09 to 0.74; p = 0.0118). These differences persisted after adjustment for age, sex, and risk factor profile differences including diabetes. Proportions of Bangladeshi and whites interpreting their symptoms as "heart attack" were similar (45.2% v 46.9%; p = 0.99). CONCLUSIONS: Bangladeshi patients with AMI often present with atypical symptoms, which may lead to slower triage in the casualty department and delay in essential treatment. This needs recognition by emergency staff if mortality rates in this high risk group are to be reduced. PMID- 12591831 TI - Multicentre evaluation of the diagnostic value of cardiac troponin T, CK-MB mass, and myoglobin for assessing patients with suspected acute coronary syndromes in routine clinical practice. AB - OBJECTIVE: To assess the diagnostic efficiency of the third generation cardiac troponin T assay in routine clinical practice. DESIGN: Prospective observational study of unselected consecutive admissions. SETTING: Multicentre study in 43 teaching and non-teaching hospitals in 13 countries. SUBJECTS: 1105 hospital admissions, median age 67 years (range 15-96 years, 63.7% male) with suspected acute coronary syndromes (72.3% of cases) or other non-specific symptoms where cardiac disease required exclusion (27.7%). INTERVENTIONS: Over the study period, myoglobin, creatine kinase MB isoenzyme (CK-MB), and cardiac troponin T where measured in parallel with conventional diagnostic tests. Final diagnostic classification involved standard ECG changes and CK-MB mass exceeding 5.0 microg/l. MAIN OUTCOME MEASURES: Diagnostic efficiency was assessed by receiver operator characteristic curve analysis including and excluding patients with unstable angina. RESULTS: Measurement of cardiac troponin T was diagnostically equivalent to CK-MB and both were better than myoglobin, with areas under the curve at 12 hours of 0.94, 0.99, and 0.80, respectively. Diagnostic criteria using CK-MB were inadequate and showed bias when patients with unstable angina were included. Elevations of cardiac troponin T did not occur when cardiac disease could be categorically excluded but were found in clinical conditions other than suspected acute coronary syndromes. CONCLUSIONS: CK-MB is unsuitable as a diagnostic gold standard even at the proposed lower threshold. A lower cut off for cardiac troponin T of 0.05 microg/l should be used for diagnosis of acute myocardial infarction. Diagnosis of acute myocardial infarction cannot be made solely on the basis of a cardiac troponin T result. PMID- 12591832 TI - Left circumflex coronary artery arising as a terminal extension of the right coronary artery. PMID- 12591833 TI - Cost effectiveness of extended treatment with low molecular weight heparin (dalteparin) in unstable coronary artery disease: results from the FRISC II trial. AB - BACKGROUND: In unstable coronary artery disease short term treatment with low molecular weight heparin in addition to aspirin has been shown to be effective. OBJECTIVE: To assess the cost effectiveness of extended treatment with dalteparin in patients managed with a non-invasive treatment strategy. DESIGN: Prospective, randomised, multicentre study. SETTING: 58 centres in Sweden, Denmark, and Norway, of which 16 were interventional. PATIENTS: After at least five days' treatment with open label dalteparin, 2267 patients were randomised to continue double blind treatment with either subcutaneous dalteparin twice daily or placebo for three months. The patients' use of health service resources was recorded prospectively. MAIN OUTCOME MEASURE: Death/myocardial infarction. RESULTS: After one month into the double blind period there was a 47% relative reduction in death or myocardial infarction in the dalteparin group compared with the placebo group (p = 0.002). There was a non-significant mean cost difference, favouring the placebo group, of 849 Swedish crowns (SEK) per patient (equivalent to 58 pounds sterling). The incremental cost effectiveness ratio for giving dalteparin treatment for one month was SEK 30 300 (range -78 000 to 139 000) (2060 pounds sterling, range -5300 pounds sterling to pound 9400 pounds sterling) per avoided death or myocardial infarct. At three months, the decrease in death or myocardial infarction was not significant, precluding cost effectiveness analyses. CONCLUSIONS: There is a marginal and non-significant increase in costs for one month of extended dalteparin treatment compared with placebo. Extended dalteparin treatment lowers the risk of death or myocardial infarction in patients with unstable coronary artery disease. While in many countries the resources for early intervention are limited, extended dalteparin treatment up to one month is a cost effective bridge to invasive intervention. PMID- 12591834 TI - Relation between heart rate, heart rhythm, and reverse left ventricular remodelling in response to carvedilol in patients with chronic heart failure: a single centre, observational study. AB - OBJECTIVE: To determine whether the process of reverse left ventricular remodelling in response to carvedilol is dependent on baseline heart rate (BHR), heart rhythm, or heart rate reduction (HRR) in response to carvedilol. DESIGN: Retrospective analysis of serial echocardiograms in 257 patients with chronic systolic heart failure at baseline and at 12-18 months after starting carvedilol. Reverse left ventricular remodelling was determined by changes in left ventricular end diastolic dimension (LVEDD), end systolic dimension (LVESD), and fractional shortening (LVFS). SETTING: Heart failure clinic within a university teaching hospital. MAIN OUTCOME MEASURES: Changes in LVEDD, LVESD, and LVFS. RESULTS: LVEDD and LVESD decreased by 2.6 (0.4) mm and 4.9 (0.5) mm, respectively (mean (SEM)), and LVFS increased by 4.3 (0.5)% (all p < 0.0001 v baseline). Simple regression revealed no significant relation between BHR or HRR and the changes in LVEDD, LVESD, or LVFS. Stratification of patients into high and low BHR groups (above and below the mean) or according to the baseline heart rhythm (sinus rhythm v atrial fibrillation) showed no differences between groups in the extent of reverse left ventricular remodelling. Improvements in left ventricular function and dimensions were associated with significant improvements in New York Heart Association functional class. CONCLUSIONS: The benefits of carvedilol in terms of reverse left ventricular remodelling and symptomatic improvement in patients with chronic heart failure are independent of BHR, heart rhythm, and the HRR that occurs in response to carvedilol. PMID- 12591836 TI - Treatment of chronic heart failure with beta adrenergic blockade beyond controlled clinical trials: the BRING-UP experience. AB - BACKGROUND: Several large controlled trials have shown that beta blockers given to patients with heart failure (New York Heart Association functional class II IV) reduce morbidity and mortality. Despite these impressive results, implementing the use of beta blockade in clinical practice appears slow and difficult. The BRING-UP study was designed to tackle this problem. OBJECTIVES: To accelerate the adoption of beta blockade in clinical practice; to provide an epidemiological estimate of the proportion of patients with heart failure suitable for this treatment in general cardiology care; and to assess effectiveness of these drugs outside the setting of clinical trials. METHODS: The design of the study and recommendations derived from available evidence on the use of beta blockers were discussed with cardiologists during regional meetings. All consecutive heart failure patients in a one month period, whether treated or not with beta blockers, were eligible for the study. In each patient, the decision to prescribe a beta blocker was a free choice for the participating physicians. All centres were provided with carvedilol, metoprolol, and bisoprolol at appropriate doses; the choice of the drug and dosage was left to the responsible clinician. All patients were followed for one year. RESULTS: 197 cardiological centres enrolled 3091 patients, 24.9% of whom were already on beta blocker treatment at baseline. beta Blockers were newly prescribed in 32.7% of cases, more often in younger and less severely ill patients. The mean daily dose of the drugs used at one year corresponded to about 70% of the maximum dose used in clinical trials. Starting treatment with beta blockers did not affect the prescription or dosage of other recommended drugs. The overall rate of beta blocker treatment increased over the year of the study from 24.9% to 49.7%. During the 12 month period, 351 deaths occurred (11.8%). In multivariate analysis, the use of beta blockers was independently associated with a better prognosis, with a relative risk of 0.60 and a lower incidence of hospital admissions for worsening heart failure. CONCLUSIONS: The implementation of beta blockers in clinical practice is feasible and could be accelerated. These drugs are associated with a lower mortality and reduced hospital admission rates, not only in clinical trials but also in the normal clinical setting. PMID- 12591837 TI - Haemodynamic and catecholamine response to simulated ventricular tachycardia in man: effect of baseline left ventricular function. AB - OBJECTIVE: To examine the effects of baseline left ventricular function on the haemodynamic and catecholamine responses to ventricular tachycardia. DESIGN: Experimental cohort study. SETTING: Cardiac catheterisation laboratory in tertiary referral centre. SUBJECTS: 24 patients (19 male, 5 female; mean (SD) age, 59 (10) years) without coronary artery disease, divided into two groups with normal or impaired left ventricular function: group A, ejection fraction > 65% (n = 10); group B, ejection fraction < 45% (n = 14). Other medical and demographic factors were similar in the two groups. INTERVENTIONS: Ventricular tachycardia was simulated with rapid pacing at 150 beats/min for 10 minutes. MAIN OUTCOME MEASURES: Arterial blood pressure; venous plasma catecholamine concentrations. RESULTS: During rapid pacing, blood pressure was lower in group B (with impaired left ventricular function) than in group A: systolic blood pressure, 102 (11) v 115 (9) mm Hg (p = 0.005); mean blood pressure, 79 (6) v 85 (6) mm Hg (p = 0.02). The ejection fraction correlated with the lowest systolic blood pressure (r = 0.64, p = 0.0006). Although the rise in adrenaline was comparable between the two groups, the rise in noradrenaline was more pronounced (p < 0.05) in patients in group B. CONCLUSION: At low rates and in selected patients, the underlying state of left ventricular function affects haemodynamic tolerance of ventricular tachycardia. Patients with impaired left ventricular function have a lower blood pressure during ventricular tachycardia, despite an exaggerated noradrenaline release. PMID- 12591839 TI - Aortic abscess. PMID- 12591838 TI - Intravascular ultrasound of the elastic pulmonary arteries: a new approach for the evaluation of primary pulmonary hypertension. AB - OBJECTIVE: To assess the structural and functional characteristics of pulmonary arteries by intravascular ultrasound (IVUS) in the setting of primary pulmonary hypertension, and to correlate the ultrasound findings with haemodynamic variables and mortality at follow up. DESIGN: Prospective observational study. SETTING: University hospital (tertiary referral centre). PATIENTS: 20 consecutive patients with primary pulmonary hypertension (16 female; mean (SD) age, 39 (14) years). METHODS: Cardiac catheterisation and simultaneous IVUS of pulmonary artery branches at baseline and after infusion of epoprostenol. RESULTS: 33 pulmonary arteries with a mean diameter of 3.91 (0.80) mm were imaged, and wall thickening was observed in all cases, 64% being eccentric. Mean wall thickness was 0.37 (0.13) mm, percentage wall area 31.0 (9.3)%, pulsatility 14.6 (4.8)%, and pulmonary/elastic strain index 449 (174) mm Hg. No correlation was observed between IVUS findings and haemodynamic variables. Epoprostenol infusion increased pulsatility by 53% and decreased the pulmonary/elastic strain index by 41% (p = 0.0001), irrespective of haemodynamic changes. At 18 (12) months follow up, nine patients had died. A reduced pulsatility and an increased pulmonary/elastic strain index were associated with increased mortality at follow up (12.0 (4.4)% v 16.4 (4.4)%, p = 0.03; 369 (67) v 546 (216) mm Hg, p = 0.02). CONCLUSIONS: IVUS demonstrated pulmonary artery wall abnormalities in all patients with primary pulmonary hypertension, mostly eccentric. The severity of the changes did not correlate with haemodynamic variables, and epoprostenol improved pulmonary vessel stiffness. There was an association between impaired pulmonary artery functional state as determined by IVUS and mortality at follow up. PMID- 12591840 TI - Epidural haematoma presenting as polymorphic ventricular tachycardia. PMID- 12591841 TI - Evaluation of intracoronary stenting by intravascular optical coherence tomography. AB - BACKGROUND: Conventional contrast cineangiography and intravascular ultrasound (IVUS) provide a limited definition of vessel microstructure and are unable to evaluate dissection, tissue prolapse, and stent apposition on a size scale less than 100 micro m. OBJECTIVE: To evaluate the use of intravascular optical coherence tomography (OCT) to assess the coronary arteries in patients undergoing coronary stenting. METHODS: OCT was employed in patients having percutaneous coronary interventions. Images were obtained before initial balloon dilatation and following stent deployment, and were evaluated for vessel dissection, tissue prolapse, stent apposition, and stent asymmetry. IVUS images were obtained before OCT, using an automatic pull back device. RESULTS: 42 stents were imaged in 39 patients without complications. Dissection, prolapse, and incomplete stent apposition were observed more often with OCT than with IVUS. Vessel dissection was identified in eight stents by OCT and two by IVUS. Tissue prolapse was identified in 29 stents by OCT and 12 by IVUS; the extent of the prolapse (mean (SD)) was 242 (156) microm by OCT and 400 (100) microm by IVUS. Incomplete stent apposition was observed in seven stents by OCT and three by IVUS. Irregular strut separation was identified in 18 stents by both OCT and IVUS. CONCLUSIONS: Intracoronary OCT for monitoring stent deployment is feasible and provides superior contrast and resolution of arterial pathology than IVUS. PMID- 12591843 TI - Acute myocardial infarction caused by gunshot wound. PMID- 12591842 TI - Prospective evaluation of the effect of an angiotensin I converting enzyme gene polymorphism on the long term risk of major adverse cardiac events after percutaneous coronary intervention. AB - OBJECTIVE: To evaluate prospectively the influence of an angiotensin I converting enzyme (ACE) gene polymorphism on long term clinical outcome of patients with established coronary artery disease treated by percutaneous coronary intervention. DESIGN AND SETTING: Prospective observational study in a university hospital. PATIENTS: Consecutive series of 1010 patients with symptomatic coronary artery disease who underwent successful coronary stent placement from November 1996 to April 1998. MAIN OUTCOME MEASURES: Long term clinical outcome was obtained and the rates of major adverse cardiac events (death, non-fatal acute myocardial infarction, unstable angina, and revascularisation) were compared according to the insertion/deletion (I/D) polymorphism of the ACE gene. RESULTS: Of the 1010 patients 29% had the DD genotype, 51% had the ID genotype, and 20% had the II genotype. All baseline clinical angiographic and procedural characteristics were identical in the three groups of patients. Event-free survival during the follow up period (median two years) was identical in patients with the II genotype compared to those with one or two D alleles. The predictors of long term survival were age, diabetes, ejection fraction, and extension of coronary artery disease. ACE genotype had no influence on the long term survival. Additional analyses assuming dominant and recessive effects of the D allele also failed to find any association; nor did the examination of low risk subgroups. CONCLUSIONS: The ACE I/D polymorphism does not influence the long term prognosis of patients with coronary disease treated by percutaneous coronary intervention, and screening patients for this gene polymorphism is not useful for secondary prevention strategies. PMID- 12591844 TI - Percutaneous closure of a false aneurysm of the superficial femoral artery under fluoroscopic guidance. PMID- 12591845 TI - Women, older persons, and ethnic minorities: factors associated with their inclusion in randomised trials of statins 1990 to 2001. PMID- 12591846 TI - Coronary angioplasty enhances platelet reactivity through von Willebrand factor release. PMID- 12591847 TI - Non-haemorrhagic cardiac tamponade caused by a horse kick. PMID- 12591848 TI - Accuracy of reporting cardiac related events during long term follow up. PMID- 12591849 TI - Malposition of ventricular pacemaker lead as an incidental finding. PMID- 12591850 TI - Role of the autonomic nervous system in vagal atrial fibrillation. PMID- 12591853 TI - The relative contribution of the sinus and AV node to heart rate variability. PMID- 12591851 TI - Short term clinical outcome of acute saddle pulmonary embolism. PMID- 12591854 TI - Relation between circulating soluble Fas ligand and subsequent ventricular remodelling following myocardial infarction. PMID- 12591855 TI - Acquired supravalvar type of left ventricular to right atrial communication following non-penetrating cardiac trauma caused by traffic accident. PMID- 12591856 TI - Contrast echocardiography. PMID- 12591857 TI - Do guidelines influence practice? PMID- 12591858 TI - Differential diagnosis of cardiogenic syncope and seizure disorders. PMID- 12591860 TI - Takayasu's arteritis: management of left main stem stenosis. AB - Takayasu arteritis is a chronic vasculitis involving the aorta and its main branches, the pulmonary arteries, and the coronary tree, and needs to be considered in a young patient with angina, in particular when pulses are absent. This case illustrates the limitations of exercise testing in diagnosing the extent of coronary artery disease and the risks associated with coronary angiography in patients with inflammatory disease in the left main stem coronary artery. It also highlights the novel use of non-invasive scanning with positron emission tomography using 18-fluorodeoxyglucose in assessing remission from this disease. Revascularisation was performed with percutaneous transluminal coronary angioplasty and stenting as an emergency procedure, but treatment of the restenosis with directional atherectomy was based on a review of the available literature. The lymphocytic alveolitis seen in this patient has not been previously described in Takayasu's disease. PMID- 12591861 TI - Artefact mimicking tachycardia during magnetic resonance imaging in a patient with an implantable loop recorder. AB - An implantable loop recorder (ILR) was implanted in a 45 year old man with recurrent syncope. A subsequent episode of injurious syncope led to performance of a cranial and shoulder magnetic resonance imaging (MRI). An artefact mimicking both wide and narrow complex tachycardias was recorded by the ILR during the shoulder MRI but not the cranial MRI. Caution should be used when interpreting the ECGs of ILRs in patients who have undergone MRI. PMID- 12591862 TI - Unexplained recurrent pericardial effusion: a lethal warning? AB - A case of a 37 year old man with cardiac angiosarcoma causing recurrent pericardial effusion, who eventually died of cardiac rupture, is presented. The diagnosis was not established until the postmortem examination despite echocardiography, pericardiocentesis, and pericardial biopsy investigations. There is neither a specific manifestation that enables early recognition nor well proven effective treatment against this disease. Accordingly, the prognosis of cardiac angiosarcoma remains grave. A high index of suspicion is recommended in patients who present with unexplained pericardial effusion. PMID- 12591863 TI - Diffusion in biofilms. PMID- 12591864 TI - Bacterial chemotaxis: a new player in response regulator dephosphorylation. PMID- 12591865 TI - CheZ-mediated dephosphorylation of the Escherichia coli chemotaxis response regulator CheY: role for CheY glutamate 89. AB - The swimming behavior of Escherichia coli at any moment is dictated by the intracellular concentration of the phosphorylated form of the chemotaxis response regulator CheY, which binds to the base of the flagellar motor. CheY is phosphorylated on Asp57 by the sensor kinase CheA and dephosphorylated by CheZ. Previous work (Silversmith et al., J. Biol. Chem. 276:18478, 2001) demonstrated that replacement of CheY Asn59 with arginine resulted in extreme resistance to dephosphorylation by CheZ despite proficient binding to CheZ. Here we present the X-ray crystal structure of CheYN59R in a complex with Mn(2+) and the stable phosphoryl analogue BeF(3)(-). The overall folding and active site architecture are nearly identical to those of the analogous complex containing wild-type CheY. The notable exception is the introduction of a salt bridge between Arg59 (on the beta3alpha3 loop) and Glu89 (on the beta4alpha4 loop). Modeling this structure into the (CheY-BeF(3)(-)-Mg(2+))(2)CheZ(2) structure demonstrated that the conformation of Arg59 should not obstruct entry of the CheZ catalytic residue Gln147 into the active site of CheY, eliminating steric interference as a mechanism for CheZ resistance. However, both CheYE89A and CheYE89Q, like CheYN59R, conferred clockwise flagellar rotation phenotypes in strains which lacked wild-type CheY and displayed considerable (approximately 40-fold) resistance to dephosphorylation by CheZ. CheYE89A and CheYE89Q had autophosphorylation and autodephosphorylation properties similar to those of wild type CheY and could bind to CheZ with wild-type affinity. Therefore, removal of Glu89 resulted specifically in CheZ resistance, suggesting that CheY Glu89 plays a role in CheZ-mediated dephosphorylation. The CheZ resistance of CheYN59R can thus be largely explained by the formation of the salt bridge between Arg59 and Glu89, which prevents Glu89 from executing its role in catalysis. PMID- 12591866 TI - Two nucleoside uptake systems in Lactococcus lactis: competition between purine nucleosides and cytidine allows for modulation of intracellular nucleotide pools. AB - A method for measuring internal nucleoside triphosphate pools of lactococci was optimized and validated. This method is based on extraction of (33)P-labeled nucleotides with formic acid and evaluation by two-dimensional chromatography with a phosphate buffer system for the first dimension and with an H(3)BO(3)-LiOH buffer for separation in the second dimension. We report here the sizes of the ribo- and deoxyribonucleotide pools in laboratory strain MG1363 during growth in a defined medium. We found that purine- and pyrimidine-requiring strains may be used to establish physiological conditions in batch fermentations with altered nucleotide pools and growth rates by addition of nucleosides in different combinations. Addition of cytidine together with inosine to a purine-requiring strain leads to a reduction in the internal purine nucleotide pools and a decreased growth rate. This effect was not seen if cytidine was replaced by uridine. A similar effect was observed if cytidine and inosine were added to a pyrimidine-requiring strain; the UTP pool size was significantly decreased, and the growth rate was reduced. To explain the observed inhibition, the nucleoside transport systems in Lactococcus lactis were investigated by measuring the uptake of radioactively labeled nucleosides. The K(m) for for inosine, cytidine, and uridine was determined to be in the micromolar range. Furthermore, it was found that cytidine and inosine are competitive inhibitors of each other, whereas no competition was found between uridine and either cytidine or inosine. These findings suggest that there are two different high-affinity nucleoside transporters, one system responsible for uridine uptake and another system responsible for the uptake of all purine nucleosides and cytidine. PMID- 12591867 TI - Dual role of cysteine 172 in redox regulation of ribulose 1,5-bisphosphate carboxylase/oxygenase activity and degradation. AB - Alkylation and oxidation of cysteine residues significantly decrease the catalytic activity and stimulate the degradation of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO). We analyzed the role of vicinal cysteine residues in redox regulation of RuBisCO from Synechocystis sp. strain PCC 6803. Cys172 and Cys192, which are adjacent to the catalytic site, and Cys247, which cross-links two large subunits, were replaced by alanine. Whereas all mutant cells (C172A, C192A, C172A-C192A, and C247A) and the wild type grew photoautotrophically at similar rates, the maximal photosynthesis rates of C172A mutants decreased 10 to 20% as a result of 40 to 60% declines in RuBisCO turnover number. Replacement of Cys172, but not replacement of Cys192, prominently decreased the effect of cysteine alkylation or oxidation on RuBisCO. Oxidants that react with vicinal thiols had a less inhibitory effect on the activity of either the C172A or C192A enzyme variants, suggesting that a disulfide bond was formed upon oxidation. Thiol oxidation induced RuBisCO dissociation into subunits. This effect was either reduced in the C172A and C192A mutant enzymes or eliminated by carboxypentitol bisphosphate (CPBP) binding to the activated enzyme form. The CPBP effect presumably resulted from a conformational change in the carbamylated CPBP-bound enzyme, as implied from an alteration in the electrophoretic mobility. Stress conditions, provoked by nitrate deprivation, decreased the RuBisCO contents and activities in the wild type and in the C192A and C247A mutants but not in the C172A and C172A-C192A mutants. These results suggest that although Cys172 does not participate in catalysis, it plays a role in redox regulation of RuBisCO activity and degradation. PMID- 12591868 TI - Synergistic effects of cellulosomal xylanase and cellulases from Clostridium cellulovorans on plant cell wall degradation. AB - Plant cell walls are comprised of cellulose and hemicellulose and other polymers that are intertwined, and this complex structure presents a barrier to degradation by pure cellulases or hemicellulases. In this study, we determined the synergistic effects on corn cell wall degradation by the action of cellulosomal xylanase XynA and cellulosomal cellulases from Clostridium cellulovorans. XynA minicellulosomes and cellulase minicellulosomes were found to degrade corn cell walls synergistically but not purified substrates such as xylan and crystalline cellulose. The mixture of XynA and cellulases at a molar ratio of 1:2 showed the highest synergistic effect of 1.6 on corn cell wall degradation. The amounts both of xylooligosaccharides and cellooligosaccharides liberated from corn cell walls were increased by the synergistic action of XynA and cellulases. Although synergistic effects on corn cell wall degradation were found in simultaneous reactions with XynA and cellulases, no synergistic effects were observed in sequential reactions. The possible mechanism of synergism between XynA and cellulases is discussed. PMID- 12591869 TI - Genetic and biochemical characterization of the F-ATPase operon from Streptococcus sanguis 10904. AB - Oral streptococci utilize an F-ATPase to regulate cytoplasmic pH. Previous studies have shown that this enzyme is a principal determinant of aciduricity in the oral streptococcal species Streptococcus sanguis and Streptococcus mutans. Differences in the pH optima of the respective ATPases appears to be the main reason that S. mutans is more tolerant of low pH values than S. sanguis and hence pathogenic. We have recently reported the genetic arrangement for the S. mutans operon. For purposes of comparative structural biology we have also investigated the F-ATPase from S. sanguis. Here, we report the genetic characterization and expression in Escherichia coli of the S. sanguis ATPase operon. Sequence analysis showed a gene order of atpEBFHAGDC and that a large intergenic space existed upstream of the structural genes. Activity data demonstrate that ATPase activity is induced under acidic conditions in both S. sanguis and S. mutans; however, it is not induced to the same extent in the nonpathogenic S. sanguis. Expression studies with an atpD deletion strain of E. coli showed that S. sanguis-E. coli hybrid enzymes were able to degrade ATP but were not sufficiently functional to permit growth on succinate minimal media. Hybrid enzymes were found to be relatively insensitive to inhibition by dicyclohexylcarbodiimide, indicating loss of productive coupling between the membrane and catalytic subunits. PMID- 12591870 TI - Efficient turnover of chlorocatechols is essential for growth of Ralstonia eutropha JMP134(pJP4) in 3-chlorobenzoic acid. AB - Ralstonia eutropha JMP134(pJP4) degrades 3-chlorobenzoate (3-CB) by using two not completely isofunctional, pJP4-encoded chlorocatechol degradation gene clusters, tfdC(I)D(I)E(I)F(I) and tfdD(II)C(II)E(II)F(II). Introduction of several copies of each gene cluster into R. eutropha JMP222, which lacks pJP4 and thus accumulates chlorocatechols from 3-CB, allows the derivatives to grow in this substrate. However, JMP222 derivatives containing one chromosomal copy of each cluster did not grow in 3-CB. The failure to grow in 3-CB was the result of accumulation of chlorocatechols due to the limiting activity of chlorocatechol 1,2-dioxygenase (TfdC), the first enzyme in the chlorocatechol degradation pathway. Micromolar concentrations of 3- and 4-chlorocatechol inhibited the growth of strains JMP134 and JMP222 in benzoate, and cells of strain JMP222 exposed to 3 mM 3-CB exhibited a 2-order-of-magnitude decrease in viability. This toxicity effect was not observed with strain JMP222 harboring multiple copies of the tfdC(I) gene, and the derivative of strain JMP222 containing tfdC(I)D(I)E(I)F(I) plus multiple copies of the tfdC(I) gene could efficiently grow in 3-CB. In addition, tfdC(I) and tfdC(II) gene mutants of strain JMP134 exhibited no growth and impaired growth in 3-CB, respectively. The introduction into strain JMP134 of the xylS-xylXYZL genes, encoding a broad-substrate-range benzoate 1,2-dioxygenase system and thus increasing the transformation of 3-CB into chlorocatechols, resulted in derivatives that exhibited a sharp decrease in the ability to grow in 3-CB. These observations indicate that the dosage of chlorocatechol-transforming genes is critical for growth in 3-CB. This effect depends on a delicate balance between chlorocatechol-producing and chlorocatechol consuming reactions. PMID- 12591871 TI - A two-component regulator of universal stress protein expression and adaptation to oxygen starvation in Mycobacterium smegmatis. AB - We identified a response regulator in Mycobacterium smegmatis which plays an important role in adaptation to oxygen-starved stationary phase. The regulator exhibits strong sequence similarity to DevR/Rv3133c of M. tuberculosis. The structural gene is present on a multigene locus, which also encodes a sensor kinase. A devR mutant of M. smegmatis was adept at surviving growth arrest initiated by either carbon or nitrogen starvation. However, its culturability decreased several orders of magnitude below that of the wild type under oxygen starved stationary-phase conditions. Two-dimensional gel analysis revealed that a number of oxygen starvation-inducible proteins were not expressed in the devR mutant. Three of these proteins are universal stress proteins, one of which is encoded directly upstream of devR. Another protein closely resembles a proposed nitroreductase, while a fifth protein corresponds to the alpha-crystallin (HspX) orthologue of M. smegmatis. None of the three universal stress proteins or nitroreductase, and a considerably lower amount of HspX was detected in carbon starved wild-type cultures. A fusion of the hspX promoter to gfp demonstrated that DevR directs gene expression when M. smegmatis enters stationary phase brought about, in particular, by oxygen starvation. To our knowledge, this is the first time a role for a two-component response regulator in the control of universal stress protein expression has been shown. Notably, the devR mutant was 10(4)-fold more sensitive than wild type to heat stress. We conclude that DevR is a stationary-phase regulator required for adaptation to oxygen starvation and resistance to heat stress in M. smegmatis. PMID- 12591873 TI - Glyceraldehyde-3-phosphate dehydrogenase has no control over glycolytic flux in Lactococcus lactis MG1363. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has previously been suggested to have almost absolute control over the glycolytic flux in Lactococcus lactis (B. Poolman, B. Bosman, J. Kiers, and W. N. Konings, J. Bacteriol. 169:5887-5890, 1987). Those studies were based on inhibitor titrations with iodoacetate, which specifically inhibits GAPDH, and the data suggested that it should be possible to increase the glycolytic flux by overproducing GAPDH activity. To test this hypothesis, we constructed a series of mutants with GAPDH activities from 14 to 210% of that of the reference strain MG1363. We found that the glycolytic flux was unchanged in the mutants overproducing GAPDH. Also, a decrease in the GAPDH activity had very little effect on the growth rate and the glycolytic flux until 25% activity was reached. Below this activity level, the glycolytic flux decreased proportionally with decreasing GAPDH activity. These data show that GAPDH activity has no control over the glycolytic flux (flux control coefficient = 0.0) at the wild-type enzyme level and that the enzyme is present in excess capacity by a factor of 3 to 4. The early experiments by Poolman and coworkers were performed with cells resuspended in buffer, i.e., nongrowing cells, and we therefore analyzed the control by GAPDH under similar conditions. We found that the glycolytic flux in resting cells was even more insensitive to changes in the GAPDH activity; in this case GAPDH was also present in a large excess and had no control over the glycolytic flux. PMID- 12591872 TI - Polymorphism in the collagen-like region of the Bacillus anthracis BclA protein leads to variation in exosporium filament length. AB - We recently identified a Bacillus anthracis glycoprotein which is a structural constituent of the exosporium filaments (P. Sylvestre, E. Couture-Tosi, and M. Mock, Mol. Microbiol. 45:169-178, 2002). This Bacillus collagen-like protein (BclA) contains an internal collagen-like region (CLR) of GXX repeats which includes a large proportion of GPT triplets. Here, we report that the polymorphic marker Ceb-Bams13, for which there are nine alleles (P. Le Fleche et al., BMC Microbiol. 1:2, 2001), maps within the open reading frame encoding BclA. The bclA gene in 11 B. anthracis strains representative of seven Ceb-Bams13 alleles was sequenced and compared to the Ames bclA gene sequence. The amino- and carboxy terminal sequences surrounding the CLR are conserved. The CLR itself is highly polymorphic: it contains between 17 and 91 GXX repeats and one to eight copies of the 21-amino-acid sequence (GPT)(5)GDTGTT, named the BclA repeat. The length of the filament on the spore surface differed between the strains. We exchanged the bclA gene between strains with different CLRs and examined the spore surfaces by electron microscopy analysis. The length of the BclA CLR is responsible for the variation in filament length. PMID- 12591874 TI - Exogenous glutathione completes the defense against oxidative stress in Haemophilus influenzae. AB - Since they are equipped with several strategies by which they evade the antimicrobial defense of host macrophages, it is surprising that members of the genus Haemophilus appear to be deficient in common antioxidant systems that are well established to protect prokaryotes against oxidative stress. Among others, no genetic evidence for glutathione (gamma-Glu-Cys-Gly) (GSH) biosynthesis or for alkyl hydroperoxide reduction (e.g., the Ahp system characteristic or enteric bacteria) is apparent from the Haemophilus influenzae Rd genome sequence, suggesting that the organism relies on alternative systems to maintain redox homeostasis or to reduce small alkyl hydroperoxides. In this report we address this apparent paradox for the nontypeable H. influenzae type strain NCTC 8143. Instead of biosynthesis, we could show that this strain acquires GSH by importing the thiol tripeptide from the growth medium. Although such GSH accumulation had no effect on growth rates, the presence of cellular GSH protected against methylglyoxal, tert-butyl hydroperoxide (t-BuOOH), and S-nitrosoglutathione toxicity and regulated the activity of certain antioxidant enzymes. H. influenzae NCTC 8143 extracts were shown to contain GSH-dependent peroxidase activity with t BuOOH as the peroxide substrate. The GSH-mediated protection against t-BuOOH stress is most probably catalyzed by the product of open reading frame HI0572 (Prx/Grx), which we isolated from a genomic DNA fragment that confers wild-type resistance to t-BuOOH toxicity in the Ahp-negative Escherichia coli strain TA4315 and that introduces GSH-dependent alkyl hydroperoxide reductase activity into naturally GSH peroxidase-negative E. coli. Finally, we demonstrated that cysteine is an essential amino acid for growth and that cystine, GSH, glutathione amide, and cysteinylglycine can be catabolized in order to complement cysteine deficiency. PMID- 12591875 TI - The Escherichia coli lipB gene encodes lipoyl (octanoyl)-acyl carrier protein:protein transferase. AB - In an earlier study (S. W. Jordan and J. E. Cronan, Jr., J. Biol. Chem. 272:17903 17906, 1997) we reported a new enzyme, lipoyl-[acyl carrier protein]-protein N lipoyltransferase, in Escherichia coli and mitochondria that transfers lipoic acid from lipoyl-acyl carrier protein to the lipoyl domains of pyruvate dehydrogenase. It was also shown that E. coli lipB mutants lack this enzyme activity, a finding consistent with lipB being the gene that encoded the lipoyltransferase. However, it remained possible that lipB encoded a positive regulator required for lipoyltransferase expression or action. We now report genetic and biochemical evidence demonstrating that lipB encodes the lipoyltransferase. A lipB temperature-sensitive mutant was shown to produce a thermolabile lipoyltransferase and a tagged version of the lipB-encoded protein was purified to homogeneity and shown to catalyze the transfer of either lipoic acid or octanoic acid from their acyl carrier protein thioesters to the lipoyl domain of pyruvate dehydrogenase. In the course of these experiments the ATG initiation codon commonly assigned to lipB genes in genomic databases was shown to produce a nonfunctional E. coli LipB protein, whereas initiation at an upstream TTG codon gave a stable and enzymatically active protein. Prior genetic results (T. W. Morris, K. E. Reed, and J. E. Cronan, Jr., J. Bacteriol. 177:1-10, 1995) suggested that lipoate protein ligase (LplA) could also utilize (albeit poorly) acyl carrier protein substrates in addition to its normal substrates lipoic acid plus ATP. We have detected a very slow LplA-catalyzed transfer of lipoic acid and octanoic acid from their acyl carrier protein thioesters to the lipoyl domain of pyruvate dehydrogenase. A nonhydrolyzable lipoyl-AMP analogue was found to competitively inhibit both ACP-dependent and ATP-dependent reactions of LplA, suggesting that the same active site catalyzes two chemically diverse reactions. PMID- 12591876 TI - Novel spoIIE mutation that causes uncompartmentalized sigmaF activation in Bacillus subtilis. AB - During sporulation, Bacillus subtilis undergoes an asymmetric division that results in two cells with different fates, the larger mother cell and the smaller forespore. The protein phosphatase SpoIIE, which is required for activation of the forespore-specific transcription factor sigma(F), is also required for optimal efficiency and timing of asymmetric division. We performed a genetic screen for spoIIE mutants that were impaired in sporulation but not sigma(F) activity and isolated a strain with the mutation spoIIEV697A. The mutant exhibited a 10- to 40-fold reduction in sporulation and a sixfold reduction in asymmetric division compared to the parent. Transcription of the sigma(F) dependent spoIIQ promoter was increased more than 10-fold and was no longer confined to the forespore. The excessive sigma(F) activity persisted even when asymmetric division was prevented. Disruption of spoIIGB did not restore asymmetric division to the spoIIEV697A mutant, indicating that the deficiency is not a consequence of predivisional activation of the mother cell-specific transcription factor sigma(E). Deletion of the gene encoding sigma(F) (spoIIAC) restored asymmetric division; however, a mutation that dramatically reduced the number of promoters responsive to sigma(F), spoIIAC561 (spoIIACV233 M), failed to do so. This result suggests that the block is due to expression of one of the small subset of sigma(F)-dependent genes expressed in this background or to unregulated interaction of sigmaF with some other factor. Our results indicate that regulation of SpoIIE plays a critical role in coupling asymmetric division to sigma(F) activation in order to ensure proper spatial and temporal expression of forespore-specific genes. PMID- 12591877 TI - Multiple light inputs control phototaxis in Synechocystis sp. strain PCC6803. AB - The phototactic behavior of individual cells of the cyanobacterium Synechocystis sp. strain PCC6803 was studied with a glass slide-based phototaxis assay. Data from fluence rate-response curves and action spectra suggested that there were at least two light input pathways regulating phototaxis. We observed that positive phototaxis in wild-type cells was a low fluence response, with peak spectral sensitivity at 645 and 704 nm. This red-light-induced phototaxis was inhibited or photoreversible by infrared light (760 nm). Previous work demonstrated that a taxD1 mutant (Cyanobase accession no. sll0041; also called pisJ1) lacked positive but maintained negative phototaxis. Therefore, the TaxD1 protein, which has domains that are similar to sequences found in both bacteriophytochrome and the methyl-accepting chemoreceptor protein, is likely to be the photoreceptor that mediates positive phototaxis. Wild-type cells exhibited negative phototaxis under high-intensity broad-spectrum light. This phenomenon is predominantly blue light responsive, with a maximum sensitivity at approximately 470 nm. A weakly negative phototactic response was also observed in the spectral region between 600 and 700 nm. A deltataxD1 mutant, which exhibits negative phototaxis even under low fluence light, has a similar action maximum in the blue region of the spectrum, with minor peaks from green to infrared (500 to 740 nm). These results suggest that while positive phototaxis is controlled by the red light photoreceptor TaxD1, negative phototaxis in Synechocystis sp. strain PCC6803 is mediated by one or more (as yet) unidentified blue light photoreceptors. PMID- 12591878 TI - Chromosomal expression of the Haemophilus influenzae Hap autotransporter allows fine-tuned regulation of adhesive potential via inhibition of intermolecular autoproteolysis. AB - The Haemophilus influenzae Hap autotransporter is a nonpilus adhesin that promotes adherence to respiratory epithelial cells and selected extracellular matrix proteins and facilitates bacterial aggregation and microcolony formation. Hap consists of a 45-kDa outer membrane translocator domain called Hap(beta) and a 110-kDa extracellular passenger domain called Hap(S). All adhesive activity resides within Hap(S), which also contains protease activity and directs its own secretion from the bacterial cell surface via intermolecular autoproteolysis. In the present study, we sought to determine the relationship between the magnitude of Hap expression, the efficiency of Hap autoproteolysis, and the level of Hap mediated adherence and aggregation. We found that a minimum threshold of Hap precursor was required for autoproteolysis and that this threshold approximated expression of Hap from a chromosomal allele, as occurs in H. influenzae clinical isolates. Chromosomal expression of wild-type Hap was sufficient to promote significant adherence to epithelial cells and extracellular matrix proteins, and adherence was enhanced substantially by inhibition of autoproteolysis. In contrast, chromosomal expression of Hap was sufficient to promote bacterial aggregation only when autoproteolysis was inhibited, indicating that the threshold for Hap-mediated aggregation is above the threshold for autoproteolysis. These results highlight the critical role of autoproteolysis and an intermolecular mechanism of cleavage in controlling the diverse adhesive activities of Hap. PMID- 12591879 TI - The cyclic AMP-cyclic AMP receptor protein complex regulates activity of the traJ promoter of the Escherichia coli conjugative plasmid pRK100. AB - The TraJ protein is a central activator of F-like plasmid conjugal transfer. In a search for regulators of traJ expression, we studied the possible regulatory role of the cyclic AMP (cAMP)-cAMP receptor protein (CRP) complex in traJ transcription using a traJ-lacZ reporter system. A comparison of the enzyme activities in the wild-type Escherichia coli strain MC4100 with those in cya and crp mutants indicated that disruption of the formation of the cAMP-CRP complex negatively influenced the activity of the traJ promoter of the F-like plasmid pRK100. The defect in the cya mutant was partially restored by addition of exogenous cAMP. Competitive reverse transcription-PCR performed with RNA isolated from the wild-type and mutant strains showed that the cAMP-CRP complex exerted its effect at the level of transcription. Electrophoretic mobility shift assays with purified CRP demonstrated that there was direct binding of CRP to the traJ promoter region. DNase I footprint experiments mapped the CRP binding site around position -67.5 upstream of the putative traJ promoter. Targeted mutagenesis of the traJ promoter region confirmed the location of the CRP binding site. Consistent with the demonstrated regulation of TraJ by the cAMP-CRP complex, mutants with defects in cya or crp exhibited reduced conjugal transfer from pRK100. PMID- 12591880 TI - Role of the cytoplasmic C terminus of the FliF motor protein in flagellar assembly and rotation. AB - Twenty-six FliF monomers assemble into the MS ring, a central motor component of the bacterial flagellum that anchors the structure in the inner membrane. Approximately 100 amino acids at the C terminus of FliF are exposed to the cytoplasm and, through the interaction with the FliG switch protein, a component of the flagellar C ring, are essential for the assembly of the motor. In this study, we have dissected the entire cytoplasmic C terminus of the Caulobacter crescentus FliF protein by high-resolution mutational analysis and studied the mutant forms with regard to the assembly, checkpoint control, and function of the flagellum. Only nine amino acids at the very C terminus of FliF are essential for flagellar assembly. Deletion or substitution of about 10 amino acids preceding the very C terminus of FliF resulted in assembly-competent but nonfunctional flagella, making these the first fliF mutations described so far with a Fla(+) but Mot(-) phenotype. Removal of about 20 amino acids further upstream resulted in functional flagella, but cells carrying these mutations were not able to spread efficiently on semisolid agar plates. At least 61 amino acids located between the functionally relevant C terminus and the second membrane-spanning domain of FliF were not required for flagellar assembly and performance. A strict correlation was found between the ability of FliF mutant versions to assemble into a flagellum, flagellar class III gene expression, and a block in cell division. Motile suppressors could be isolated for nonmotile mutants but not for mutants lacking a flagellum. Several of these suppressor mutations were localized to the 5' region of the fliG gene. These results provide genetic support for a model in which only a short stretch of amino acids at the immediate C terminus of FliF is required for flagellar assembly through stable interaction with the FliG switch protein. PMID- 12591881 TI - Synthesis of a Klebsiella pneumoniae O-antigen heteropolysaccharide (O12) requires an ABC 2 transporter. AB - A recombinant clone encoding enzymes for Klebsiella pneumoniae O12-antigen lipopolysaccharide (LPS) was found when we screened for serum resistance of a cosmid-based genomic library of K. pneumoniae KT776 (O12:K80) introduced into Escherichia coli DH5alpha. A total of eight open reading frames (ORFs) (wb(O12) gene cluster) were necessary to produce K. pneumoniae O12-antigen LPS in E. coli K-12. A complete analysis of the K. pneumoniae wb(O12) cluster revealed an interesting coincidence with the wb(O4) cluster of Serratia marcescens from ORF5 to ORF8 (or WbbL to WbbA). This prompted us to generate mutants of K. pneumoniae strain KT776 (O12) and to study complementation between the two enterobacterial wb clusters using mutants of S. marcescens N28b (O4) obtained previously. Both wb gene clusters are examples of ABC 2 transporter-dependent pathways for O-antigen heteropolysaccharides. The wzm-wzt genes and the wbbA or wbbB genes were not interchangeable between the two gene clusters despite their high level of similarity. However, introduction of three cognate genes (wzm-wzt-wbbA or wzm-wzt wbbB) into mutants unable to produce O antigen allowed production of the specific O antigen. The K. pneumoniae O12 WbbL protein performs the same function as WbbL from S. marcescens O4 in either the S. marcescens O4 or E. coli K-12 genetic background. PMID- 12591882 TI - Rhamnogalacturonate lyase RhiE is secreted by the out system in Erwinia chrysanthemi. AB - Supernatants of rhamnose-induced Erwinia chrysanthemi strain 3937 cultures contain a principal secreted protein named RhiE. A rhiE mutant has been found among a set of rhamnose-induced MudI1681 lacZ fusions. RhiE is a 62-kDa protein that has rhamnogalacturonate lyase activity on rhamnogalacturonan I (RG-I). It does not require a divalent cation for its activity and has an optimal pH of 6.0. rhiE expression is strongly induced in the presence of rhamnose but is also regulated by PecT and Crp, two regulators of the transcription of pectinolytic enzyme genes. RhiE is secreted through the type II Out secretion pathway. RhiE has no disulfide bond. The absence of RhiE secretion in a dsb mutant indicated that disulfide bond formation is required for the biogenesis of the secretion apparatus. RhiE was searched for in several E. chrysanthemi strains by using antibodies, and it was found to be present in one-third of the strains tested. However, the reduced virulence of the rhiE mutant indicates that degradation of the RG-I region of pectin is important for full virulence of E. chrysanthemi. PMID- 12591883 TI - Functional characterization of penicillin-binding protein 1b from Streptococcus pneumoniae. AB - The widespread use of antibiotics has encouraged the development of drug resistance in pathogenic bacteria. In order to overcome this problem, the modification of existing antibiotics and/or the identification of targets for the design of new antibiotics is currently being undertaken. Bifunctional penicillin binding proteins (PBPs) are membrane-associated molecules whose transpeptidase (TP) activity is irreversibly inhibited by beta-lactam antibiotics and whose glycosyltransferase (GT) activity represents a potential target in the antibacterial fight. In this work, we describe the expression and the biochemical characterization of the soluble extracellular region of Streptococcus pneumoniae PBP1b (PBP1b*). The acylation efficiency for benzylpenicillin and cefotaxime was characterized by stopped-flow fluorometry and a 40-kDa stable TP domain was generated after limited proteolysis. In order to analyze the GT activity of PBP1b*, we developed an electrophoretic assay which monitors the fluorescence signal from PBP1b*-bound dansylated lipid II. This binding was inhibited by the antibiotic moenomycin and was specific for the GT domain, since no signal was observed in the presence of the purified functional TP domain. Binding studies performed with truncated forms of PBP1b* demonstrated that the first conserved motif of the GT domain is not required for the recognition of lipid II, whereas the second motif is necessary for such interaction. PMID- 12591884 TI - Overexpression of the waaZ gene leads to modification of the structure of the inner core region of Escherichia coli lipopolysaccharide, truncation of the outer core, and reduction of the amount of O polysaccharide on the cell surface. AB - The waa gene cluster is responsible for the biosynthesis of the lipopolysaccharide (LPS) core region in Escherichia coli and Salmonella: Homologs of the waaZ gene product are encoded by the waa gene clusters of Salmonella enterica and E. coli strains with the K-12 and R2 core types. Overexpression of WaaZ in E. coli and S. enterica led to a modified LPS structure showing core truncations and (where relevant) to a reduction in the amount of O-polysaccharide side chains. Mass spectrometry and nuclear magnetic resonance spectroscopy were used to determine the predominant LPS structures in an E. coli isolate with an R1 core (waaZ is lacking from the type R1 waa gene cluster) with a copy of the waaZ gene added on a plasmid. Novel truncated LPS structures, lacking up to 3 hexoses from the outer core, resulted from WaaZ overexpression. The truncated molecules also contained a KdoIII residue not normally found in the R1 core. PMID- 12591885 TI - Complex regulation of the Bacillus subtilis aconitase gene. AB - The roles of the CcpC, CodY, and AbrB proteins in regulation of the Bacillus subtilis aconitase (citB) gene were found to be distinct and to vary with the conditions and phase of growth. CcpC, a citrate-inhibited repressor that is the primary factor regulating citB expression in minimal-glucose-glutamine medium, also contributed to repression of citB during exponential-phase growth in broth medium. A null mutation in codY had no effect on citB expression during growth in minimal medium even when combined with ccpC and abrB mutations. However, a codY mutation slightly relieved repression during exponential growth in broth medium and completely derepressed citB expression when combined with a ccpC mutation. An abrB mutation led to decreased expression of citB during stationary phase in both broth and minimal medium. All three proteins bound in vitro to specific and partially overlapping sites within the citB regulatory region. Interaction of CcpC and CodY with the citB promoter region was partially competitive. PMID- 12591886 TI - Conjugative transfer of p42a from rhizobium etli CFN42, which is required for mobilization of the symbiotic plasmid, is regulated by quorum sensing. AB - Rhizobium etli CFN42 contains six plasmids. Only one of them, p42a, is self conjugative at high frequency. This plasmid is strictly required for mobilization of the symbiotic plasmid (pSym). To study the transfer mechanism of p42a, a self transmissible cosmid clone containing its transfer region was isolated. Its sequence showed that most of the tra genes are highly similar to genes of Agrobacterium tumefaciens pTiC58 and other related plasmids. Four putative regulatory genes were identified; three of these (traI, traR, and cinR) belong to the LuxR-LuxI family. Mutagenesis of these genes confirmed their requirement for p42a transfer. We found that the conjugative transfer of p42a is dependent on quorum sensing, and consequently pSym transfer also was found to be similarly regulated, establishing a complex link between environmental conditions and pSym transfer. Although R. etli has been shown to produce different N-acyl-homoserine lactones, only one of them, a 3-oxo-C(8)-homoserine lactone encoded by the traI gene described here, was involved in transfer. Mutagenesis of the fourth regulatory gene, traM, had no effect on transfer. Analysis of transcriptional fusions of the regulatory genes to a reporter gene suggests a complex regulation scheme for p42a conjugative transfer. Conjugal transfer gene expression was found to be directly upregulated by TraR and the 3-oxo-C(8)-homoserine lactone synthesized by TraI. The traI gene was autoregulated by these elements and positively regulated by CinR, while cinR expression required traI. Finally, we did not detect expression of traM, indicating that in p42a TraM may be expressed so weakly that it cannot inhibit conjugal transfer, leading to the unrepressed transfer of p42a. PMID- 12591887 TI - P15 and P3, the tail completion proteins of bacteriophage T4, both form hexameric rings. AB - Two proteins, gp15 and gp3 (gp for gene product), are required to complete the assembly of the T4 tail. gp15 forms the connector which enables the tail to bind to the head, whereas gp3 is involved in terminating the elongation of the tail tube. In this work, genes 15 and 3 were cloned and overexpressed, and the purified gene products were studied by analytical ultracentrifugation, electron microscopy, and circular dichroism. Determination of oligomerization state by sedimentation equilibrium revealed that both gp15 and gp3 are hexamers of the respective polypeptide chains. Electron microscopy of the negatively stained P15 and P3 (P denotes the oligomeric state of the gene product) revealed that both proteins form hexameric rings, the diameter of which is close to that of the tail tube. The differential roles between gp15 and gp3 upon completion of the tail are discussed. PMID- 12591888 TI - The Escherichia coli methyl-directed mismatch repair system repairs base pairs containing oxidative lesions. AB - A major role of the methyl-directed mismatch repair (MMR) system of Escherichia coli is to repair postreplicative errors. In this report, we provide evidence that MMR also acts on oxidized DNA, preventing mutagenesis. When cells deficient in MMR are grown anaerobically, spontaneous mutation frequencies are reduced compared with those of the same cells grown aerobically. In addition, we show that a dam mutant has an increased sensitivity to hydrogen peroxide treatment that can be suppressed by mutations that inactivate MMR. In a dam mutant, MMR is not targeted to newly replicated DNA strands and therefore mismatches are converted to single- and double-strand DNA breaks. Thus, base pairs containing oxidized bases will be converted to strand breaks if they are repaired by MMR. This is demonstrated by the increased peroxide sensitivity of a dam mutant and the finding that the sensitivity can be suppressed by mutations inactivating MMR. We demonstrate further that this repair activity results from MMR recognition of base pairs containing 8-oxoguanine (8-oxoG) based on the finding that overexpression of the MutM oxidative repair protein, which repairs 8-oxoG, can suppress the mutH-dependent increase in transversion mutations. These findings demonstrate that MMR has the ability to prevent oxidative mutagenesis either by removing 8-oxoG directly or by removing adenine misincorporated opposite 8-oxoG or both. PMID- 12591889 TI - Characterization of a cytosolic NiFe-hydrogenase from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. AB - We have identified an NiFe-hydrogenase exclusively localized in the cytoplasm of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 (T. kodakaraensis hydrogenase). A gene cluster encoding T. kodakaraensis hydrogenase was composed of four open reading frames (hyhBGSL(Tk)), where the hyhS(Tk) and hyhL(Tk) gene products corresponded to the small and the large subunits of NiFe-hydrogenase, respectively. A putative open reading frame for hydrogenase-specific maturation endopeptidase (hybD(Tk)) was found downstream of the cluster. Polyclonal antibodies raised against recombinant HyhL(Tk) were used for immunoaffinity purification of T. kodakaraensis hydrogenase, leading to a 259-fold concentration of hydrogenase activity. The purified T. kodakaraensis hydrogenase was composed of four subunits (beta, gamma, delta, and alpha), corresponding to the products of hyhBGSL(Tk), respectively. Each alphabetagammadelta unit contained 0.8 mol of Ni, 22.3 mol of Fe, 21.1 mol of acid-labile sulfide, and 1.01 mol of flavin adenine dinucleotide. The optimal temperature for the T. kodakaraensis hydrogenase was 95 degrees C for H(2) uptake and 90 degrees C for H(2) production with methyl viologen as the electron carrier. We found that NADP(+) and NADPH promoted high levels of uptake and evolution of H(2), respectively, suggesting that the molecule is the electron carrier for the T. kodakaraensis hydrogenase. PMID- 12591890 TI - Structural model for 12-helix transporters belonging to the major facilitator superfamily. AB - The major facilitator superfamily includes a large collection of evolutionarily related proteins that have been implicated in the transport of a variety of solutes and metabolites across the membranes of organisms ranging from bacteria to humans. We have recently reported the three-dimensional structure, at 6.5 A resolution, of the oxalate transporter, OxlT, a representative member of this superfamily. In the oxalate-bound state, 12 helices surround a central cavity to form a remarkably symmetrical structure that displays a well-defined pseudo twofold axis perpendicular to the plane of the membrane as well as two less pronounced, mutually perpendicular pseudo twofold axes in the plane of the membrane. Here, we combined this structural information with sequence information from other members of this protein family to arrive at models for the arrangement of helices in this superfamily of transport proteins. Our analysis narrows down the number of helix arrangements from about a billion starting possibilities to a single probable model for the relative spatial arrangement for the 12 helices, consistent both with our structural findings and with the majority of previous biochemical studies on members of this superfamily. PMID- 12591891 TI - DNA microarray analysis of redox-responsive genes in the genome of the cyanobacterium Synechocystis sp. strain PCC 6803. AB - Whole-genome DNA microarrays were used to evaluate the effect of the redox state of the photosynthetic electron transport chain on gene expression in Synechocystis sp. strain PCC 6803. Two specific inhibitors of electron transport, 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and 2,5-dibromo-3-methyl-6 isopropyl-p-benzoquinone (DBMIB), were added to the cultures, and changes in accumulation of transcripts were examined. About 140 genes were highlighted as reproducibly affected by the change in the redox state of the photosynthetic electron transport chain. It was shown that some stress-responsive genes but not photosynthetic genes were under the control of the redox state of the plastoquinone pool in Synechocystis sp. strain PCC 6803. PMID- 12591892 TI - tRNA2Thr complements temperature sensitivity caused by null mutations in the htrB gene in Escherichia coli. AB - According to the wobble rule, tRNA2Thr is nonessential for protein synthesis, because the codon (ACG) that is recognized by tRNA2Thr is also recognized by tRNA4Thr. In order to investigate the reason that this nonessential tRNA nevertheless exists in Escherichia coli, we attempted to isolate tRNA2Thr requiring mutants. Using strain JM101F(-), which lacks the gene for tRNA2Thr, we succeeded in isolating two temperature-sensitive mutants whose temperature sensitivity was complemented by introduction of the gene for tRNA2Thr. These mutants had a mutation in the htrB gene, whose product is an enzyme involved in lipid A biosynthesis. Although it is known that some null mutations in the htrB gene give a temperature-sensitive phenotype, our mutants exhibited tighter temperature sensitivity. We discuss a possible mechanism for the requirement for tRNA2Thr. PMID- 12591893 TI - Mechanism of cis-trans isomerization of unsaturated fatty acids in Pseudomonas putida. AB - We studied the pattern of the cis-trans isomerization of unsaturated fatty acids in cells of Pseudomonas putida S12 grown in a medium supplemented with oleic acid which was deuterated at both of the C atoms of its double bond. Direct evidence that isomerization does not include a transient saturation of the double bond was obtained. In addition, analysis of the amino acid sequences of the seven known Cti proteins identified them as heme-containing proteins of the cytochrome c type. PMID- 12591894 TI - A suppressor of the menadione-hypersensitive phenotype of a Xanthomonas campestris pv. phaseoli oxyR mutant reveals a novel mechanism of toxicity and the protective role of alkyl hydroperoxide reductase. AB - We isolated menadione-resistant mutants of Xanthomonas campestris pv. phaseoli oxyR (oxyR(Xp)). The oxyRR2(Xp) mutant was hyperresistant to the superoxide generators menadione and plumbagin and was moderately resistant to H(2)O(2) and tert-butyl hydroperoxide. Analysis of enzymes involved in oxidative-stress protection in the oxyRR2(Xp) mutant revealed a >10-fold increase in AhpC and AhpF levels, while the levels of superoxide dismutase (SOD), catalase, and the organic hydroperoxide resistance protein (Ohr) were not significantly altered. Inactivation of ahpC in the oxyRR2(Xp) mutant resulted in increased sensitivity to menadione killing. Moreover, high levels of expression of cloned ahpC and ahpF in the oxyR(Xp) mutant complemented the menadione hypersensitivity phenotype. High levels of other oxidant-scavenging enzymes such as catalase and SOD did not protect the cells from menadione toxicity. These data strongly suggest that the toxicity of superoxide generators could be mediated via organic peroxide production and that alkyl hydroperoxide reductase has an important novel function in the protection against the toxicity of these compounds in X. campestris. PMID- 12591895 TI - Hemin binding, functional expression, and complementation analysis of Pap 31 from Bartonella henselae. AB - Growth of Bartonella henselae is strongly heme dependent, and B. henselae is unable to synthesize heme itself. At least five outer membrane-associated proteins from B. henselae bind hemin, including the 31-kDa protein designated Pap31. The gene of this protein was heterologously expressed in Escherichia coli M15(pREP4) and detected with monoclonal antibodies in the outer membrane fraction. Complementation of the hemA-deficient mutant E. coli K-12 EB53 (aroB tsx malT hemA) with pap31 demonstrated that this protein is involved in heme acquisition and may be an important virulence factor in the pathogenesis of B. henselae. PMID- 12591896 TI - Transposition of Tn5367 in Mycobacterium marinum, using a conditionally recombinant mycobacteriophage. AB - Mycobacterium marinum is a close relative of the obligate human pathogen Mycobacterium tuberculosis. As with M. tuberculosis, M. marinum causes intracellular infection of poikilothermic vertebrates and skin infection in humans. It is considered a valid model organism for the study of intracellular pathogenesis of mycobacteria. Low transformation efficiencies for this species have precluded approaches using mutant libraries in pathogenesis studies. We have adapted the conditionally replicating mycobacteriophage phAE94, originally developed as a transposon mutagenesis tool for M. tuberculosis, to meet the specific requirements of M. marinum. Conditions permissive for phage replication in M. tuberculosis facilitated highly efficient transposon delivery in M. marinum. Using this technique we succeeded in generating a representative mutant library of this species, and we conclude that TM4-derived mycobacteriophages are temperature-independent suicide vectors for M. marinum. PMID- 12591897 TI - Molecular cloning of endo-beta-D-1,4-glucanase genes, rce1, rce2, and rce3, from Rhizopus oryzae. AB - Three endoglucanase genes, designated the rce1, rce2, and rce3 genes, were isolated from Rhizopus oryzae as the first cellulase genes from the subdivision ZYGOMYCOTA: All the amino acid sequences deduced from the rce1, rce2, and rce3 genes consisted of three distinct domains: cellulose binding domains, linker domains, and catalytic domains belonging to glycosyl hydrolase family 45. The rce3 gene had two tandem repeated sequences of cellulose binding domains, while rce1 and rce2 had only one. rce1, rce2, and rce3 had various lengths of linker sequences. PMID- 12591898 TI - Control of immune responses by naturally arising CD4+ regulatory T cells that express toll-like receptors. PMID- 12591899 TI - Regulatory T cells selectively express toll-like receptors and are activated by lipopolysaccharide. AB - Regulatory CD4 T cells (Treg) control inflammatory reactions to commensal bacteria and opportunist pathogens. Activation of Treg functions during these processes might be mediated by host-derived proinflammatory molecules or directly by bacterial products. We tested the hypothesis that engagement of germline encoded receptors expressed by Treg participate in the triggering of their function. We report that the subset of CD4 cells known to exert regulatory functions in vivo (CD45RB(low) CD25(+)) selectively express Toll-like receptors (TLR)-4, -5, -7, and -8. Exposure of CD4(+) CD25(+) cells to the TLR-4 ligand lipopolysaccharide (LPS) induces up-regulation of several activation markers and enhances their survival/proliferation. This proliferative response does not require antigen-presenting cells and is augmented by T cell receptor triggering and interleukin 2 stimulation. Most importantly, LPS treatment increases CD4(+) CD25(+) cell suppressor efficiency by 10-fold and reveals suppressive activity in the CD4(+) CD45RB(low) CD25(-) subset that when tested ex-vivo, scores negative. Moreover, LPS-activated Treg efficiently control naive CD4 T cell-dependent wasting disease. These findings provide the first evidence that Treg respond directly to proinflammatory bacterial products, a mechanism that likely contributes to the control of inflammatory responses. PMID- 12591900 TI - Changes in chromatin accessibility across the GM-CSF promoter upon T cell activation are dependent on nuclear factor kappaB proteins. AB - Granulocyte/macrophage colony-stimulating factor (GM-CSF) is a key cytokine in myelopoiesis and aberrant expression is associated with chronic inflammatory disease and myeloid leukemias. This aberrant expression is often associated with constitutive nuclear factor (NF)-kappaB activation. To investigate the relationship between NF-kappaB and GM-CSF transcription in a chromatin context, we analyzed the chromatin structure of the GM-CSF gene in T cells and the role of NF-kappaB proteins in chromatin remodeling. We show here that chromatin remodeling occurs across a region of the GM-CSF gene between -174 and +24 upon T cell activation, suggesting that remodeling is limited to a single nucleosome encompassing the proximal promoter. Nuclear NF-kappaB levels appear to play a critical role in this process. In addition, using an immobilized template assay we found that the ATPase component of the SWI/SNF chromatin remodeling complex, brg1, is recruited to the GM-CSF proximal promoter in an NF-kappaB-dependent manner in vitro. These results suggest that chromatin remodeling across the GM CSF promoter in T cells is a result of recruitment of SWI/SNF type remodeling complexes by NF-kappaB proteins binding to the CD28 response region of the promoter. PMID- 12591901 TI - A role of suppressor of cytokine signaling 3 (SOCS3/CIS3/SSI3) in CD28-mediated interleukin 2 production. AB - Suppressor of cytokine signaling (SOCS)3 has been characterized as a negative feedback regulator in cytokine-mediated Janus kinase signal transducer and activator of transcription signaling. However, this study shows that T cells from transgenic mice expressing SOCS3 exhibit a significant reduction in interleukin (IL)-2 production induced by T cell receptor cross-linking when T cells are costimulated with CD28. Decreased protein expression in SOCS3(+/-) mice enhanced CD28-mediated IL-2 production, clearly indicating the correlation between expression level of SOCS3 and IL-2 production ability. The SOCS3 protein interacted with phosphorylated CD28 through its SH2 domain but not the kinase inhibitory region. In addition, a point mutation in the SOCS3 SH2 domain attenuated the inhibition of CD28 function in IL-2 promoter activation. Committed T helper (Th)2 cells exclusively expressed SOCS3 and production of Th2 cytokines, such as IL-4 and IL-5, was much less dependent on CD28 costimulation compared with interferon gamma and IL-2 production in Th1 cells. Consistent with this notion, the expression level of SOCS3 in early T cell activation influenced the ability of IL-2 production induced by CD28 costimulation. Therefore, the SOCS3 may play an alternative role in prohibiting excessive progression of CD28 mediated IL-2 production. PMID- 12591902 TI - Selective activation of the c-Jun NH2-terminal protein kinase signaling pathway by stimulatory KIR in the absence of KARAP/DAP12 in CD4+ T cells. AB - Activation of CD4(+) T cells is governed by interplay between stimulatory and inhibitory receptors; predominance of stimulatory signals favors autoimmune reactions. In patients with rheumatoid arthritis, expression of the critical costimulatory molecule, CD28, is frequently lost. Instead, CD4(+)CD28(null) T cells express killer immunoglobulin-like receptors (KIRs) with a preferential expression of the stimulatory receptor, CD158j. The frequency of CD4(+)CD28(null) T cells in rheumatoid arthritis (RA) correlates with the risk for more severe disease. Moreover, the KIR2DS2 gene, which encodes for CD158j, is a genetic risk factor for rheumatoid vasculitis. CD158j signals through the adaptor molecule, KARAP/DAP12, to positively regulate cytotoxic activity in NK cells. However, the majority of CD4(+)CD28(null) T cell clones lacked the expression of KARAP/DAP12. Despite the absence of KARAP/DAP12, CD158j was functional and augmented interferon-gamma production after T cell receptor stimulation. Cross-linking of CD158j resulted in selective phosphorylation of c-Jun NH(2)-terminal protein kinase (JNK) and its upstream kinase, MKK4 that led to the expression of ATF-2 and c-Jun, all in the absence of extracellular signal-regulated kinase (ERK)1/2 phosphorylation. Mutation of the lysine residue within the transmembrane domain of CD158j abolished JNK activation, suggesting that an alternate adaptor molecule was being used. CD4(+)CD28(null) T cells expressed DAP10 and inhibition of phosphatidylinositol 3-kinase, which acts downstream of DAP10, inhibited JNK activation; however, no interaction of DAP10 with CD158j could be detected. Our data suggest that CD158j in T cells functions as a costimulatory molecule through the JNK pathway independent of KARAP/DAP12 and DAP10. Costimulation by CD158j may contribute to the autoreactivity of CD4(+)CD28(null) T cells in RA. PMID- 12591903 TI - T cell regulation as a side effect of homeostasis and competition. AB - We have previously hypothesized that maintaining a balanced peripheral immune system may not be the sole responsibility of a specialized subset of T cells dedicated to immune regulation, but also a side effect of normal competition for shared resources within an intact immune system. Here we show that regulatory activity is correlated with high homeostatic expansion potential, reflecting the avidity for self-peptide:MHC complexes. Monoclonal transgenic T cells with high homeostatic expansion potential and lacking characteristics previously associated with regulatory function were able to regulate wasting disease induced by transfer of a small number of naive CD45RB(hi) CD4 T cells into lymphopenic hosts. Self-regulatory function is also found in the naive polyclonal T cell repertoire depleted of CD25(+) T cells. T cells capable of preventing immune pathology, like the transgenic T cells, express higher than average levels of CD5, an indicator of avidity for self:MHC peptide complexes. We therefore propose that dysregulated expansion of potentially pathogenic T cells in a lymphopenic environment can be prevented by members of the naive T cell repertoire, irrespective of their specificity, as a side effect of their response to homeostatic and antigenic stimulation. PMID- 12591904 TI - Sustained activation of cell adhesion is a differentially regulated process in B lymphopoiesis. AB - It is largely unknown how hematopoietic progenitors are positioned within specialized niches of the bone marrow microenvironment during development. Chemokines such as CXCL12, previously called stromal cell-derived factor 1, are known to activate cell integrins of circulating leukocytes resulting in transient adhesion before extravasation into tissues. However, this short-term effect does not explain the mechanism by which progenitor cells are retained for prolonged periods in the bone marrow. Here we show that in human bone marrow CXCL12 triggers a sustained adhesion response specifically in progenitor (pro- and pre-) B cells. This sustained adhesion diminishes during B cell maturation in the bone marrow and, strikingly, is absent in circulating mature B cells, which exhibit only transient CXCL12-induced adhesion. The duration of adhesion is tightly correlated with CXCL12-induced activation of focal adhesion kinase (FAK), a known molecule involved in integrin-mediated signaling. Sustained adhesion of progenitor B cells is associated with prolonged FAK activation, whereas transient adhesion in circulating B cells is associated with short-lived FAK activation. Moreover, sustained and transient adhesion responses are differentially affected by pharmacological inhibitors of protein kinase C and phosphatidylinositol 3 kinase. These results provide a developmental cell stage-specific mechanism by which chemokines orchestrate hematopoiesis through sustained rather than transient activation of adhesion and cell survival pathways. PMID- 12591905 TI - In vitro evidence that cytokine receptor signals are required for differentiation of double positive thymocytes into functionally mature CD8+ T cells. AB - CD4(+)8(+) double positive (DP) thymocytes differentiate into CD4(+) and CD8(+) mature T cells in response to TCR signals. However, TCR signals that are initiated in DP thymocytes are unlikely to persist throughout all subsequent differentiation steps, suggesting that other signals must sustain thymocyte differentiation after TCR signaling has ceased. Using an in vitro experimental system, we now demonstrate that cytokine receptor signals, such as those transduced by IL-7 receptors, are required for differentiation of signaled DP thymocytes into functionally mature CD8(+) T cells as they: (a) up-regulate Bcl-2 expression to maintain thymocyte viability; (b) enhance CD4 gene silencing; (c) promote functional maturation;and (d) up-regulate surface expression of glucose transporter molecules, which improve nutrient uptake and increase metabolic activity. IL-7Rs appear to be unique among cytokine receptors in maintaining the viability of newly generated CD4(-)8(+) thymocytes, whereas several different cytokine receptors can provide the trophic/differentiative signals for subsequent CD8(+) thymocyte differentiation and maturation. Thus, cytokine receptors provide both survival and trophic/differentiative signals with varying degrees of redundancy that are required for differentiation of signaled DP thymocytes into functionally mature CD8(+) T cells. PMID- 12591906 TI - Prevention of arthritis by interleukin 10-producing B cells. AB - In this study we have shown that activation of arthritogenic splenocytes with antigen and agonistic anti-CD40 gives raise to a B cell population that produce high levels of interleukin (IL)-10 and low levels of interferon (IFN)-gamma. Transfer of these B cells into DBA/1-TcR-beta-Tg mice, immunized with bovine collagen (CII) emulsified in complete Freund's adjuvant inhibited T helper type 1 differentiation, prevented arthritis development, and was also effective in ameliorating established disease. IL-10 is essential for the regulatory function of this subset of B cells, as the B cells population isolated from IL-10 knockout mice failed to mediate this protective function. Furthermore, B cells isolated from arthritogenic splenocytes treated in vitro with anti-IL-10/anti-IL-10R were unable to protect recipient mice from developing arthritis. Our results suggest a new role of a subset of B cells in controlling T cell differentiation and autoimmune disorders. PMID- 12591907 TI - A mouse with a loss-of-function mutation in the c-Cbl TKB domain shows perturbed thymocyte signaling without enhancing the activity of the ZAP-70 tyrosine kinase. AB - The unique tyrosine kinase binding (TKB) domain of Cbl targets phosphorylated tyrosines on activated protein tyrosine kinases (PTKs); this targeting is considered essential for Cbl proteins to negatively regulate PTKs. Here, a loss of-function mutation (G304E) in the c-Cbl TKB domain, first identified in Caenorhabditis elegans, was introduced into a mouse and its effects in thymocytes and T cells were studied. In marked contrast to the c-Cbl knockout mouse, we found no evidence of enhanced activity of the ZAP-70 PTK in thymocytes from the TKB domain mutant mouse. This finding contradicts the accepted mechanism of c-Cbl mediated negative regulation, which requires TKB domain targeting of phosphotyrosine 292 in ZAP-70. However, the TKB domain mutant mouse does show aspects of enhanced signaling that parallel those of the c-Cbl knockout mouse, but these involve the constitutive activation of Rac and not enhanced PTK activity. Furthermore, the enhanced signaling in CD4(+)CD8(+) double positive thymocytes appears to be compensated by the selective down-regulation of CD3 on mature thymocytes and peripheral T cells from both strains of mutant c-Cbl mice. PMID- 12591908 TI - Transgenic expression of the activating natural killer receptor Ly49H confers resistance to cytomegalovirus in genetically susceptible mice. AB - Natural resistance to infection with mouse cytomegalovirus (MCMV) is controlled by a dominant locus, Cmv1. Cmv1 is linked to the Ly49 family of natural killer receptors on distal chromosome 6. While some studies localized Cmv1 as distal to the Ly49 gene cluster, genetic and functional analysis identified Ly49h as a pivotal factor in resistance to MCMV. The role of these two independent genomic domains in MCMV resistance was evaluated by functional complementation using transgenesis of bacterial artificial chromosomes (BAC) in genetically susceptible mice. Phenotypic and genetic characterization of the transgenic animals traced the resistance gene to a single region spanning the Ly49h gene. The appearance of the Ly49H protein in NK cells of transgenic mice coincided with the emergence of MCMV resistance, and there was a threshold Ly49H protein level associated with full recovery. Finally, transgenic expression of Ly49H in the context of either of the two independent susceptibility alleles, Cmv1(sBALB) or Cmv1(sFVB), conferred resistance to MCMV infection. These results demonstrate that Ly49h is necessary and sufficient to confer MCMV resistance, and formally demonstrate allelism between Cmv1 and Ly49h. This panel of transgenic animals provides a unique resource to study possible pleiotropic effect of Cmv1. PMID- 12591910 TI - Interleukin 1 receptor dependence of serum transferred arthritis can be circumvented by toll-like receptor 4 signaling. AB - Inflammatory arthritis is associated with the release of a network of key cytokines. In T cell receptor transgenic K/BxN mice interleukin (IL)-1 plays a key role in joint swelling and destruction, as suggested by the ability of anti IL-1receptor (IL-1R) antibody treatment to delay the onset and slow the progression of this disease. This mechanism is dependent on the signaling pathway intermediary myeloid differentiation factor 88 (MyD88), such that neither IL-1R nor MyD88-deficient mice developed visually detectable synovitis after transfer of arthritogenic sera. The Toll-like receptors (TLRs) share the same signaling pathway through MyD88 as the IL-1R. The administration of a TLR-4 ligand, lipopolysaccharide, concomitant with arthritogenic serum in IL-1 receptor deficient mice resulted in acute paw swelling, but not in MyD88-deficient mice. Also, serum transferred arthritis was not sustained in TLR-4 mutant mice compared with controls. These results suggest that innate immune functions via TLR-4 might perpetuate inflammatory mechanisms and bypass the need for IL-1 in chronic joint inflammation. PMID- 12591909 TI - Low penetrance, broad resistance, and favorable outcome of interleukin 12 receptor beta1 deficiency: medical and immunological implications. AB - The clinical phenotype of interleukin 12 receptor beta1 chain (IL-12Rbeta1) deficiency and the function of human IL-12 in host defense remain largely unknown, due to the small number of patients reported. We now report 41 patients with complete IL-12Rbeta1 deficiency from 17 countries. The only opportunistic infections observed, in 34 patients, were of childhood onset and caused by weakly virulent Salmonella or Mycobacteria (Bacille Calmette-Guerin -BCG- and environmental Mycobacteria). Three patients had clinical tuberculosis, one of whom also had salmonellosis. Unlike salmonellosis, mycobacterial infections did not recur. BCG inoculation and BCG disease were both effective against subsequent environmental mycobacteriosis, but not against salmonellosis. Excluding the probands, seven of the 12 affected siblings have remained free of case-definition opportunistic infection. Finally, only five deaths occurred in childhood, and the remaining 36 patients are alive and well. Thus, a diagnosis of IL-12Rbeta1 deficiency should be considered in children with opportunistic mycobacteriosis or salmonellosis; healthy siblings of probands and selected cases of tuberculosis should also be investigated. The overall prognosis is good due to broad resistance to infection and the low penetrance and favorable outcome of infections. Unexpectedly, human IL-12 is redundant in protective immunity against most microorganisms other than Mycobacteria and Salmonella. Moreover, IL-12 is redundant for primary immunity to Mycobacteria and Salmonella in many individuals and for secondary immunity to Mycobacteria but not to Salmonella in most. PMID- 12591911 TI - Is all of the endoplasmic reticulum created equal? The effects of the heterogeneous distribution of endoplasmic reticulum Ca2+-handling proteins. AB - The endoplasmic reticulum is a heterogeneous compartment with respect to the distribution of its Ca2+-handling proteins, namely the Ca2+-binding proteins, the Ca2+ pumps and the Ca2+ release channels. The nonuniform distribution of these proteins may explain the functional heterogeneity of the endoplasmic reticulum, such as the generation of spatially complex Ca2+ signals, Ca2+ homeostasis, and protein folding and quality control. PMID- 12591912 TI - Steady-state dynamics of Cajal body components in the Xenopus germinal vesicle. AB - Cajal bodies (CBs) are evolutionarily conserved nuclear organelles that contain many factors involved in the transcription and processing of RNA. It has been suggested that macromolecular complexes preassemble or undergo maturation within CBs before they function elsewhere in the nucleus. Most such models of CB function predict a continuous flow of molecules between CBs and the nucleoplasm, but there are few data that directly support this view. We used fluorescence recovery after photobleaching (FRAP) on isolated Xenopus oocyte nuclei to measure the steady-state exchange rate between the nucleoplasm and CBs of three fluorescently tagged molecules: U7 small nuclear RNA, coilin, and TATA-binding protein (TBP). In the nucleoplasm, the apparent diffusion coefficients for the three molecules ranged from 0.26 to 0.40 microm2 s-1. However, in CBs, fluorescence recovery was markedly slower than in the nucleoplasm, and there were at least three kinetic components. The recovery rate within CBs was independent of bleach spot diameter and could not be attributed to high CB viscosity or density. We propose that binding to other molecules and possibly assembly into larger complexes are the rate-limiting steps for FRAP of U7, coilin, and TBP inside CBs. PMID- 12591913 TI - The molecular function of Ase1p: evidence for a MAP-dependent midzone-specific spindle matrix. Microtubule-associated proteins. AB - The midzone is the domain of the mitotic spindle that maintains spindle bipolarity during anaphase and generates forces required for spindle elongation (anaphase B). Although there is a clear role for microtubule (MT) motor proteins at the spindle midzone, less is known about how microtubule-associated proteins (MAPs) contribute to midzone organization and function. Here, we report that budding yeast Ase1p is a member of a conserved family of midzone-specific MAPs. By size exclusion chromatography and velocity sedimentation, both Ase1p in extracts and purified Ase1p behaved as a homodimer. Ase1p bound and bundled MTs in vitro. By live cell microscopy, loss of Ase1p resulted in a specific defect: premature spindle disassembly in mid-anaphase. Furthermore, when overexpressed, Ase1p was sufficient to trigger spindle elongation in S phase-arrested cells. FRAP revealed that Ase1p has both a very slow rate of turnover within the midzone and limited lateral diffusion along spindle MTs. We propose that Ase1p functions as an MT cross-bridge that imparts matrix-like characteristics to the midzone. MT dependent networks of spindle midzone MAPs may be one molecular basis for the postulated spindle matrix. PMID- 12591914 TI - Characterization of the translocon of the outer envelope of chloroplasts. AB - The protein translocon of the outer envelope of chloroplasts (Toc) consists of the core subunits Toc159, Toc75, and Toc34. To investigate the molecular structure, the core complex was purified. This core complex has an apparent molecular mass of approximately 500 kD and a molecular stoichiometry of 1:4:4-5 between Toc159, Toc75, and Toc34. The isolated translocon recognizes both transit sequences and precursor proteins in a GTP-dependent manner, suggesting its functional integrity. The complex is embedded by the lipids phosphatidylcholine and digalactosyldiacylglyceride. Two-dimensional structural analysis by EM revealed roughly circular particles consistent with the formation of a stable core complex. The particles show a diameter of approximately 130 A with a solid ring and a less dense interior structure. A three-dimensional map obtained by random conical tilt reconstruction of electron micrographs suggests that a "finger"-like central region separates four curved translocation channels within one complex. PMID- 12591916 TI - Compartmentalized megakaryocyte death generates functional platelets committed to caspase-independent death. AB - Caspase-directed apoptosis usually fragments cells, releasing nonfunctional, prothrombogenic, membrane-bound apoptotic bodies marked for rapid engulfment by macrophages. Blood platelets are functional anucleate cells generated by specialized fragmentation of their progenitors, megakaryocytes (MKs), but committed to a constitutive caspase-independent death. Constitutive formation of the proplatelet-bearing MK was recently reported to be caspase-dependent, apparently involving mitochondrial release of cytochrome c, a known pro apoptogenic factor. We extend those studies and report that activation of caspases in MKs, either constitutively or after Fas ligation, yields platelets that are functionally responsive and evade immediate phagocytic clearance, and retain mitochondrial transmembrane potential until constitutive platelet death ensues. Furthermore, the exclusion from the platelet progeny of caspase-9 present in the progenitor accounts for failure of mitochondrial release of cytochrome c to activate caspase-3 during platelet death. Thus, progenitor cell death by apoptosis can result in birth of multiple functional anucleate daughter cells. PMID- 12591915 TI - The inner membrane protein Mdm33 controls mitochondrial morphology in yeast. AB - Mitochondrial distribution and morphology depend on MDM33, a Saccharomyces cerevisiae gene encoding a novel protein of the mitochondrial inner membrane. Cells lacking Mdm33 contain ring-shaped, mostly interconnected mitochondria, which are able to form large hollow spheres. On the ultrastructural level, these aberrant organelles display extremely elongated stretches of outer and inner membranes enclosing a very narrow matrix space. Dilated parts of Delta mdm33 mitochondria contain well-developed cristae. Overexpression of Mdm33 leads to growth arrest, aggregation of mitochondria, and generation of aberrant inner membrane structures, including septa, inner membrane fragments, and loss of inner membrane cristae. The MDM33 gene is required for the formation of net-like mitochondria in mutants lacking components of the outer membrane fission machinery, and mitochondrial fusion is required for the formation of extended ring-like mitochondria in cells lacking the MDM33 gene. The Mdm33 protein assembles into an oligomeric complex in the inner membrane where it performs homotypic protein-protein interactions. Our results indicate that Mdm33 plays a distinct role in the mitochondrial inner membrane to control mitochondrial morphology. We propose that Mdm33 is involved in fission of the mitochondrial inner membrane. PMID- 12591917 TI - Amassin, an olfactomedin protein, mediates the massive intercellular adhesion of sea urchin coelomocytes. AB - Sea urchins have a fluid-filled body cavity, the coelom, containing four types of immunocytes called coelomocytes. Within minutes after coelomic fluid is removed from the body cavity, a massive cell-cell adhesion of coelomocytes occurs. This event is referred to as clotting. Clotting is thought to be a defense mechanism against loss of coelomic fluid if the body wall is punctured, and it may also function in the cellular encapsulation of foreign material and microbes. Here we show that this intercoelomocyte adhesion is mediated by amassin, a coelomic plasma protein with a relative molecular mass (Mr) of 75 kD. Amassin forms large disulfide-bonded aggregates that adhere coelomocytes to each other. One half of the amassin protein comprises an olfactomedin (OLF) domain. Structural predictions show that amassin and other OLF domain-containing vertebrate proteins share a common architecture. This suggests that other proteins of the OLF family may function in intercellular adhesion. These findings are the first to demonstrate a function for a protein of the OLF family. PMID- 12591919 TI - Peroxisome proliferator-activated receptor gamma-mediated differentiation: a mutation in colon cancer cells reveals divergent and cell type-specific mechanisms. AB - Activation of the nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARgamma) inhibits cell growth and induces differentiation in both adipocyte and epithelial cell lineages, although it is unclear whether this occurs through common or cell-type specific mechanisms. We have identified four human colon cancer cell lines that do no undergo growth inhibition or induce markers of differentiation after exposure to PPARgamma agonists. Sequence analysis of the PPARgamma gene revealed that all four cell lines contain a previously unidentified point mutation in the ninth alpha-helix of the ligand binding domain at codon 422 (K422Q). The mutant receptor did not exhibit any defects in DNA binding or retinoid X receptor heterodimerization and was transcriptionally active in an artificial reporter assay. However, only retroviral transduction of the wild-type (WT), but not mutant, receptor could restore PPARgamma ligand-induced growth inhibition and differentiation in resistant colon cancer cell lines. In contrast, there was no difference in the ability of fibroblast cells expressing WT or K422Q mutant receptor to undergo growth inhibition, express adipocyte differentiation markers, or uptake lipid after treatment with a PPARgamma agonist. Finally, analysis of direct PPARgamma target genes in colon cancer cells expressing the WT or K422Q mutant allele suggests that the mutation may disrupt the ability of PPARgamma to repress the basal expression of a subset of genes in the absence of exogenous ligand. Collectively, these data argue that codon 422 may be a part of a co-factor(s) interaction domain necessary for PPARgamma to induce terminal differentiation in epithelial, but not adipocyte, cell lineages and argues that the receptor induces growth inhibition and differentiation via cell lineage-specific mechanisms. PMID- 12591918 TI - Elevated glucose inhibits VEGF-A-mediated endocardial cushion formation: modulation by PECAM-1 and MMP-2. AB - Atrioventricular (AV) septal defects resulting from aberrant endocardial cushion (EC) formation are observed at increased rates in infants of diabetic mothers. EC formation occurs via an epithelial-mesenchymal transformation (EMT), involving transformation of endocardial cells into mesenchymal cells, migration, and invasion into extracellular matrix. Here, we report that elevated glucose inhibits EMT by reducing myocardial vascular endothelial growth factor A (VEGF A). This effect is reversed with exogenous recombinant mouse VEGF-A165, whereas addition of soluble VEGF receptor-1 blocks EMT. We show that disruption of EMT is associated with persistence of platelet endothelial cell adhesion molecule-1 (PECAM-1) and decreased matrix metalloproteinase-2 (MMP-2) expression. These findings correlate with retention of a nontransformed endocardial sheet and lack of invasion. The MMP inhibitor GM6001 blocks invasion, whereas explants from PECAM-1 deficient mice exhibit MMP-2 induction and normal EMT in high glucose. PECAM-1-negative endothelial cells are highly motile and express more MMP-2 than do PECAM-1-positive endothelial cells. During EMT, loss of PECAM-1 similarly promotes single cell motility and MMP-2 expression. Our findings suggest that high glucose-induced inhibition of AV cushion morphogenesis results from decreased myocardial VEGF-A expression and is, in part, mediated by persistent endocardial cell PECAM-1 expression and failure to up-regulate MMP-2 expression. PMID- 12591920 TI - Iron regulatory protein 2 as iron sensor. Iron-dependent oxidative modification of cysteine. AB - Iron regulatory protein 2 coordinates cellular regulation of iron metabolism by binding to iron responsive elements in mRNA. The protein is synthesized constitutively but is rapidly degraded when iron stores are replete. This iron dependent degradation requires the presence of a 73-residue degradation domain, but its functions have not yet been established. We now show that the domain can act as an iron sensor, mediating its own covalent modification. The domain forms an iron-binding site with three cysteine residues located in the middle of the domain. It then reacts with molecular oxygen to generate a reactive oxidizing species at the iron-binding site. One cysteine residue is oxidized to dehydrocysteine and other products. This covalent modification may thus mark the protein molecule for degradation by the proteasome system. PMID- 12591921 TI - Resistance to beta-lactam antibiotics and its mediation by the sensor domain of the transmembrane BlaR signaling pathway in Staphylococcus aureus. AB - Staphylococci, a leading cause of infections worldwide, have devised two mechanisms for resistance to beta-lactam antibiotics. One is production of beta lactamases, hydrolytic resistance enzymes, and the other is the expression of penicillin-binding protein 2a (PBP 2a), which is not susceptible to inhibition by beta-lactam antibiotics. The beta-lactam sensor-transducer (BlaR), an integral membrane protein, binds beta-lactam antibiotics on the cell surface and transduces the information to the cytoplasm, where gene expression is derepressed for both beta-lactamase and penicillin-binding protein 2a. The gene for the sensor domain of the sensor-transducer protein (BlaR(S)) of Staphylococcus aureus was cloned, and the protein was purified to homogeneity. It is shown that beta lactam antibiotics covalently modify the BlaR(S) protein. The protein was shown to contain the unusual carboxylated lysine that activates the active site serine residue for acylation by the beta-lactam antibiotics. The details of the kinetics of interactions of the BlaR(S) protein with a series of beta-lactam antibiotics were investigated. The protein undergoes acylation by beta-lactam antibiotics with microscopic rate constants (k(2)) of 1-26 s(-1), yet the deacylation process was essentially irreversible within one cell cycle. The protein undergoes a significant conformational change on binding with beta-lactam antibiotics, a process that commences at the preacylation complex and reaches its full effect after protein acylation has been accomplished. These conformational changes are likely to be central to the signal transduction events when the organism is exposed to the beta-lactam antibiotic. PMID- 12591922 TI - The transport of low density lipoprotein-derived cholesterol to the plasma membrane is defective in NPC1 cells. AB - Niemann-Pick disease type C (NPC) is characterized by lysosomal storage of cholesterol and gangliosides, which results from defects in intracellular lipid trafficking. Most studies of NPC1 have focused on its role in intracellular cholesterol movement. Our hypothesis is that NPC1 facilitates the egress of cholesterol from late endosomes, which are where active NPC1 is located. When NPC1 is defective, cholesterol does not exit late endosomes; instead, it is carried along to lysosomal storage bodies, where it accumulates. In this study, we addressed whether cholesterol is transported from endosomes to the plasma membrane before reaching NPC1-containing late endosomes. Our study was conducted in Chinese hamster ovary cell lines that display the classical NPC biochemical phenotype and belong to the NPC1 complementation group. We used three approaches to test whether low density lipoprotein (LDL)-derived cholesterol en route to NPC1-containing organelles passes through the plasma membrane. First, we used cyclodextrins to measure the arrival of LDL cholesterol at the plasma membrane and found that the arrival of LDL cholesterol in a cyclodextrin-accessible pool was significantly delayed in NPC1 cells. Second, the movement of LDL cholesterol to NPC1-containing late endosomes was assessed and found to be normal in Chinese hamster ovary mutant 3-6, which exhibits defective movement of plasma membrane cholesterol to intracellular membranes. Third, we examined the movement of plasma membrane cholesterol to the endoplasmic reticulum and found that this pathway is intact in NPC1 cells, i.e. it does not pass through NPC1-containing late endosomes. Our data suggest that in NPC1 cells LDL cholesterol traffics directly through endosomes to lysosomes, bypassing the plasma membrane, and is trapped there because of dysfunctional NPC1. PMID- 12591923 TI - Characterization of phosphopeptide motifs specific for the Src homology 2 domains of signal transducer and activator of transcription 1 (STAT1) and STAT3. AB - Signal transducers and activators of transcription (STAT) 1 and STAT3 are activated by overlapping but distinct sets of cytokines. STATs are recruited to the different cytokine receptors through their Src homology (SH) 2 domains that make highly specific interactions with phosphotyrosine-docking sites on the receptors. We used a degenerate phosphopeptide library synthesized on 35-microm TentaGel beads and fluorescence-activated bead sorting to determine the sequence specificity of the peptide-binding sites of the SH2 domains of STAT1 and STAT3. The large bead library allowed not only peptide sequencing of pools of beads but also of single beads. The method was validated through surface plasmon resonance measurements of the affinities of different peptides to the STAT SH2 domains. Furthermore, when selected peptides were attached to a truncated erythropoietin receptor and stably expressed in DA3 cells, activation of STAT1 or STAT3 could be achieved by stimulation with erythropoietin. The combined analysis of pool sequencing, the individual peptide sequences, and plasmon resonance measurements allowed the definition of SH2 domain binding motifs. STAT1 preferentially binds peptides with the motif phosphotyrosine-(aspartic acid/glutamic acid) (proline/arginine)-(arginine/proline/glutamine), whereby a negatively charged amino acid at +1 excludes a proline at +2 and vice versa. STAT3 preferentially binds peptides with the motif phosphotyrosine-(basic or hydrophobic)-(proline or basic)-glutamine. For both STAT1 and STAT3, specific high affinity phosphopeptides were identified that can be used for the design of inhibitory molecules. PMID- 12591924 TI - Deletion of P1 arginine in a novel antithrombin variant (antithrombin London) abolishes inhibitory activity but enhances heparin affinity and is associated with early onset thrombosis. AB - A novel variant of antithrombin, the major serpin inhibitor of coagulation proteases, has been identified in a patient with early onset thrombosis and abnormal plasma antithrombin activity. Sequencing of the antithrombin genes of the patient revealed that one of the two alleles was abnormal due to an in-frame deletion of the codon for the P1 arginine residue. The abnormal antithrombin was separated from the normal inhibitor by complexing the latter with thrombin followed by heparin-agarose affinity chromatography. The purified variant, antithrombin London, was completely inactive as a thrombin or factor Xa inhibitor even after heparin activation. Surprisingly, the variant bound heparin with a K(D) reflecting an approximately 10-fold greater affinity than the normal inhibitor. Stopped-flow kinetic analysis showed that this was almost entirely due to a more favorable conformational activation of the variant than the normal inhibitor, as reflected by a decreased rate constant for reversal of the activation. Consistent with its higher than normal heparin affinity, the inactive antithrombin variant was a potent competitive antagonist of the heparin-catalyzed reaction of normal antithrombin with thrombin but did not affect the uncatalyzed reaction. These results suggest that deletion of the antithrombin P1 residue partially activates the serpin by inducing strain in the reactive center loop, which destabilizes the native loop-buried state and favors the activated loop exposed state with high heparin affinity. The unusually severe thrombosis associated with the heterozygous mutation may be explained by the ability of antithrombin London to bind endogenous heparan sulfate or heparin molecules with high affinity and to thereby block activation of the normal inhibitor. PMID- 12591925 TI - Compensatory phosphorylation and protein-protein interactions revealed by loss of function and gain of function mutants of multiple serine phosphorylation sites in endothelial nitric-oxide synthase. AB - We examined the influence of individual serine phosphorylation sites in endothelial nitric-oxide synthase (eNOS) on basal and stimulated NO release, cooperative phosphorylation, and co-association with hsp90 and Akt. Mutation of the serine phosphorylation sites 116, 617, and 1179 to alanines affected the phospho-state of at least one other site, demonstrating cooperation between multiple phosphorylation events, whereas mutation of serine 635 to alanine did not cause compensation. Mutation of serines 116 and 617 to alanine promoted a greater protein-protein interaction with hsp90 and Akt and greater phosphorylation on serine 1179, the major site for Akt phosphorylation. More importantly, using alanine substitutions, Ser-116 is important for agonist, but not basal NO release, Ser-635 is important for basal, but not stimulated, Ser-617 negatively regulates basal and stimulated NO release, and Ser-1179 phosphorylation is stimulatory for both basal and agonist-mediated NO release. Using putative "gain of function" mutants (serine to aspartate) serines 635 and 1179 are important positive regulators of basal and stimulated NO release. S635D eNOS is the most efficacious, yielding 5-fold increases in basal and 2-fold increases in stimulated NO release from cells. However, S617A and S617D eNOS both increased NO release with opposite actions in NOS activity assays. Thus, multiple serine phosphorylation events regulate basal and stimulate NO release with Ser 635 and Ser-1179 being important positive regulatory sites and Ser-116 as a negative regulatory. Ser-617 may not be important for directly regulating NO release but is important as a modulator of phosphorylation at other sites and protein-protein interactions. PMID- 12591926 TI - Tumor necrosis factor (TNF)-induced germinal center kinase-related (GCKR) and stress-activated protein kinase (SAPK) activation depends upon the E2/E3 complex Ubc13-Uev1A/TNF receptor-associated factor 2 (TRAF2). AB - Tumor necrosis factor (TNF)-induced activation of apoptosis signal-regulating kinase 1 (ASK1) and germinal center kinases (GCKs) and the subsequent activation of stress-activated protein kinases (SAPKs and c-Jun NH(2)-terminal kinases) requires TNF receptor-associated factor 2 (TRAF2). Although the TRAF2 TRAF domain binds ASK1, GCK, and the highly related kinase GCKR, the RING finger domain is needed for their activation. Here, we report that TNF activates GCKR and the SAPK pathway in a manner that depends upon TRAF2 and Ubc13, a member along with Uev1A of a dimeric ubiquitin-conjugating enzyme complex. Interference with Ubc13 function or expression inhibits both TNF- and TRAF2-mediated GCKR and SAPK activation, but has a minimal effect on ASK1 activation. TNF signaling leads to TRAF2 polyubiquitination and oligomerization and to the oligomerization, ubiquitination, and activation of GCKR, all of which are sensitive to the disruption of Ubc13 function. These results indicate that the assembly of a TRAF2 lysine 63-linked polyubiquitin chain by Ubc13/Uev1A is required for TNF-mediated GCKR and SAPK activation, but may not be required for ASK1 activation. PMID- 12591927 TI - Concomitant recruitment of ERK1/2 and p38 MAPK signalling pathway is required for activation of cytoplasmic phospholipase A2 via ATP in articular chondrocytes. AB - Extracellular ATP is a pro-inflammatory mediator involved in the release of prostaglandin from articular chondrocytes, but little is known about its effects on intracellular signaling. ATP triggered the rapid release of prostaglandin E(2) (PGE(2)) by acting on P2Y(2) receptors in rabbit articular chondrocytes. We have explored the signaling events involved in this synthesis. ATP significantly increased arachidonic acid production, which involved the activation of the 85 kDa cytosolic phospholipase A(2) (cPLA(2)) but not a secreted form of PLA(2), as demonstrated by various PLA(2) inhibitors and translocation experiments. We also showed that ATP induced the phosphorylation of p38 and ERK1/2 mitogen-activated protein kinases (MAPKs). Both PD98059, an inhibitor of the ERK pathway, and SB203580, an inhibitor of p38 MAPK, completely inhibited the ATP-induced release of PGE(2). Finally, dominant-negative plasmids encoding p38 and ERK transfected alone into the cells impaired the ATP-induced release of PGE(2) to about the same extent as both plasmids transfected together. These results suggest that PGE(2) production induced by ATP requires the activation of both ERK1/2 and p38 MAPKs. Thus, ATP acts via P2Y(2)-purine receptors to recruit cPLA(2) by activating both ERK1/2 and p38 MAPKs and stimulates the release of PGE(2) from articular chondrocytes. PMID- 12591928 TI - p53 mediates repression of the BRCA2 promoter and down-regulation of BRCA2 mRNA and protein levels in response to DNA damage. AB - Adriamycin and other DNA-damaging agents have been shown to reduce BRCA2 mRNA levels in breast cancer cell lines, but the mechanism by which this occurs is unknown. In this study, we show that adriamycin and mitomycin C, but not other DNA-damaging agents, repress BRCA2 promoter activity in a dose- and time dependent manner. We demonstrate that the effect is dependent on wild type p53 and that adriamycin and p53 mediate repression of the BRCA2 promoter by inhibiting binding of an upstream stimulatory factor protein complex to the promoter. In addition, we present evidence indicating that adriamycin and other DNA-damaging agents reduce BRCA2 mRNA and protein levels by altering both BRCA2 mRNA stability and protein stability. Thus, BRCA2 levels in the cell are regulated by three independent mechanisms in a p53-dependent manner. PMID- 12591929 TI - Live cell analysis and targeting of the lipid droplet-binding adipocyte differentiation-related protein. AB - Neutral lipid is stored in spherical organelles called lipid droplets that are bounded by a coat of proteins. The protein that is most frequently found at the surface of lipid droplets is adipocyte differentiation-related protein (ADRP). In this study, we demonstrate that fusion of either the human or mouse ADRP coding sequences to green fluorescent protein (GFP) does not disrupt the ability of the protein to associate with lipid droplets. Using this system to identify targeting elements, discontinuous segments within the coding region were required for directing ADRP to lipid droplets. GFP-tagged protein was employed also to examine the behavior of lipid droplets in live cells. Time lapse microscopy demonstrated that in HuH-7 cells, which are derived from a human hepatoma, a small number of lipid droplets could move rapidly, indicating transient association with intracellular transport pathways. Most lipid droplets did not show such movement but oscillated within a confined area; these droplets were in close association with the endoplasmic reticulum membrane and moved in concert with the endoplasmic reticulum. Fluorescence recovery analysis of GFP-tagged ADRP in live cells revealed that surface proteins do not rapidly diffuse between lipid droplets, even in conditions where they are closely packed. This system provides new insights into the properties of lipid droplets and their interaction with cellular processes. PMID- 12591930 TI - Heparan sulfate proteoglycans as regulators of fibroblast growth factor-2 signaling in brain endothelial cells. Specific role for glypican-1 in glioma angiogenesis. AB - Fibroblast growth factor-2 (FGF2) is a potent angiogenic factor in gliomas. Heparan sulfate promotes ligand binding to receptor tyrosine kinase and regulates signaling. The goal of this study was to examine the contribution of heparan sulfate proteoglycans (HSPGs) to glioma angiogenesis. Here we show that all brain endothelial cell HSPGs carry heparan sulfate chains similarly capable of forming a ternary complex with FGF2 and fibroblast growth factor receptor-1c and of promoting a mitogenic signal. Immunohistochemical analysis revealed that glypican 1 was overexpressed in glioma vessel endothelial cells, whereas this cell-surface HSPG was consistently undetectable in normal brain vessels. To determine the effect of increased glypican-1 expression on FGF2 signaling, we transfected normal brain endothelial cells, which express low base-line levels of glypican-1, with this proteoglycan. Glypican-1 expression enhanced growth of brain endothelial cells and sensitized them to FGF2-induced mitogenesis despite the fact that glypican-1 remained a minor proteoglycan. In contrast, overexpression of syndecan-1 had no effect on growth or FGF2 sensitivity. We conclude that the glypican-1 core protein has a specific role in FGF2 signaling. Glypican-1 overexpression may contribute to angiogenesis and the radiation resistance characteristic of this malignancy. PMID- 12591931 TI - Inactivation of Na,K-ATPase following Co(NH3)4ATP binding at a low affinity site in the protomeric enzyme unit. AB - The Na(+)-dependent or E1 stages of the Na,K-ATPase reaction require a few micromolar ATP, but submillimolar concentrations are needed to accelerate the K(+)-dependent or E2 half of the cycle. Here we use Co(NH(3))(4)ATP as a tool to study ATP sites in Na,K-ATPase. The analogue inactivates the K(+) phosphatase activity (an E2 partial reaction) and the Na,K-ATPase activity in parallel, whereas ATP-[(3)H]ADP exchange (an E1 reaction) is affected less or not at all. Although the inactivation occurs as a consequence of low affinity Co(NH(3))(4)ATP binding (K(D) approximately 0.4-0.6 mm), we can also measure high affinity equilibrium binding of Co(NH(3))(4)[(3)H]ATP (K(D) = 0.1 micro m) to the native enzyme. Crucially, we find that covalent enzyme modification with fluorescein isothiocyanate (which blocks E1 reactions) causes little or no effect on the affinity of the binding step preceding Co(NH(3))(4)ATP inactivation and only a 20% decrease in maximal inactivation rate. This suggests that fluorescein isothiocyanate and Co(NH(3))(4)ATP bind within different enzyme pockets. The Co(NH(3))(4)ATP enzyme was solubilized with C(12)E(8) to a homogeneous population of alphabeta protomers, as verified by analytical ultracentrifugation; the solubilization did not increase the Na,K-ATPase activity of the Co(NH(3))(4)ATP enzyme with respect to parallel controls. This was contrary to the expectation for a hypothetical (alphabeta)(2) membrane dimer with a single ATP site per protomer, with or without fast dimer/protomer equilibrium in detergent solution. Besides, the solubilized alphabeta protomer could be directly inactivated by Co(NH(3))(4)ATP, to less than 10% of the control Na,K-ATPase activity. This suggests that the inactivation must follow Co(NH(3))(4)ATP binding at a low affinity site in every protomeric unit, thus still allowing ATP and ADP access to phosphorylation and high affinity ATP sites. PMID- 12591932 TI - Antisense down-regulation of lipocalin-interacting membrane receptor expression inhibits cellular internalization of lipocalin-1 in human NT2 cells. AB - There is increasing experimental evidence demonstrating that many lipocalins bind to specific cell surface receptors. However, whereas the binding of lipocalins to their lipophilic ligands has now been characterized in much detail, there is a lack of knowledge about the nature of lipocalin receptors, the physiological role of receptor binding, and the molecular mechanism of ligand delivery. We previously identified a novel human membrane protein (lipocalin-1-interacting membrane receptor (LIMR)), which interacts with lipocalin-1 (Wojnar, P., Lechner, M., Merschak, P., and Redl, B. (2001) J. Biol. Chem. 276, 20206-20212). In the present study, we investigated the physiological role of LIMR and found this protein to be essential for mediating internalization of lipocalin-1 (Lcn-1) in NT2 cells, leading to its degradation. Whereas control NT2 cells rapidly internalized (125)I-Lcn-1 or fluorescein isothiocyanate-labeled Lcn-1, NT2 cells that were made LIMR deficient by cDNA antisense expression greatly accumulated Lcn-1 in the culture medium but did not internalize it. Because sequence and structure analysis indicated that proteins similar to LIMR are present in several organisms and at least two closely related orthologues are found in human and mouse, we suggest LIMR to be the prototype of a new family of endocytic receptors, which are topographically characterized by nine putative transmembrane domains and a characteristic large central cytoplasmic loop. PMID- 12591933 TI - A residue in the S2 subsite controls substrate selectivity of matrix metalloproteinase-2 and matrix metalloproteinase-9. AB - Matrix metalloproteinase (MMP)-2 and MMP-9 are closely related metalloproteinases that are implicated in angiogenesis. The two proteins have a similar domain structure and highly homologous catalytic domains, making them an excellent comparative model for understanding the structural basis of substrate recognition by the MMP family. Although the two MMPs exhibit some overlap in substrate recognition, our recent work showed that MMP-2 can cleave a set of peptide substrates that are only poorly recognized by MMP-9 (Chen, E. I., Kridel, S. J., Howard, E. W., Li, W., Godzik, A., and Smith, J. W. (2002) J. Biol. Chem. 277, 4485-4491). Mutations at the P(2) position of these peptide substrates dramatically reduced their selectivity for MMP-2. Inspection of the corresponding S(2) pocket of the substrate-binding cleft of the protease reveals that MMP-9 contains an Asp, whereas MMP-2 contains Glu. Here, we test the hypothesis that this conservative substitution has a role in substrate selectivity. Mutation of Glu(412) in MMP-2 to Asp significantly reduced the hydrolysis of selective substrates, with only a minor effect on hydrolysis of non-selective substrates. The predominant effect of the mutation is at the level of k(cat), or turnover rate, with reductions reaching as high as 37-fold. The residues that occupy this position in other MMPs are highly variable, providing a potential structural basis for substrate recognition across the MMP family. PMID- 12591934 TI - Subcellular localization and in vivo subunit interactions of ubiquitous mu calpain. AB - Ubiquitously expressed calpains are Ca(2+)-dependent, intracellular cysteine proteases comprising a large catalytic subunit (domains DI-DIV) and a noncovalently bound small regulatory subunit (domains DV and DVI). It is unclear whether Ca(2+)-induced calpain activation is followed by subunit dissociation or not. Here, we have applied advanced fluorescence microscopy techniques to study calpain subunit interactions in living cells using recombinant calpain subunits or domains fused to enhanced cyan and enhanced yellow fluorescent reporter proteins. All of the overexpressed variants of the catalytic subunit (DI-IV, DI III, and DI-IIb) were active and Ca(2+)-dependent. The intact large subunit, but not its truncated variants, associates with the small subunit under resting and ionomycin-activated conditions. All of the variants were localized in cytoplasm and nuclei, except DI-IIb, which accumulates in the nucleus and in nucleoli as shown by microscopy and cell fractionation. Localization studies with mutated and chimeric variants indicate that nuclear targeting of the DI-IIb variant is conferred by the two N-terminal helices of DI. Only those variants that contain DIII migrated to membranes upon the addition of ionomycin, suggesting that DIII is essential for membrane targeting. We propose that intracellular localization and in particular membrane targeting of activated calpain, but not dissociation of its intact subunits, contribute to regulate its proteolytic activity in vivo. PMID- 12591935 TI - Notch-induced proteolysis and nuclear localization of the Delta ligand. AB - The Delta protein is a single-pass transmembrane ligand for the Notch family of receptors. Delta binding to Notch invokes regulated intramembrane proteolysis and nuclear translocation of the Notch intracellular domain. Delta is proteolytically processed at two sites, Ala(581) and Ala(593) in the juxtamembrane and transmembrane domains, respectively (Mishra-Gorur, K., Rand, M. D., Perez Villamil, B., and Artavanis-Tsakonas, S. (2002) J. Cell Biol. 159, 313-324). Controversy over the role of Delta processing in propagating Notch signals has stemmed from conflicting reports on the activity or inactivity of soluble extracellular domain products of Delta. We have examined Delta proteolysis in greater detail and report that Delta undergoes three proteolytic cleavages in the region of the juxtamembrane and transmembrane domains. Only one of these cleavages, analogous to cleavage at Ala(581), is dependent on the Kuzbanian ADAM metalloprotease. The two additional cleavages correspond to the previously described cleavage at Ala(593) and a novel unidentified site within or close to the transmembrane domain. Delta processing is up-regulated in co-cultures with Notch-expressing cells and is similarly induced by p-aminophenylmercuric acetate, a well documented activator of metalloproteases. Furthermore, expression of a truncated intracellular isoform of Delta shows prominent nuclear localization. Altogether, these data demonstrate a role for Notch in inducing Delta proteolysis and implicate a nuclear function for Delta, consistent with a model of bi directional signaling through Notch-Delta interactions. PMID- 12591936 TI - Relaxed sugar donor selectivity of a Sinorhizobium meliloti ortholog of the Rhizobium leguminosarum mannosyl transferase LpcC. Role of the lipopolysaccharide core in symbiosis of Rhizobiaceae with plants. AB - The lpcC gene of Rhizobium leguminosarum and the lpsB gene of Sinorhizobium meliloti encode protein orthologs that are 58% identical over their entire lengths of about 350 amino acid residues. LpcC and LpsB are required for symbiosis with pea and Medicago plants, respectively. S. meliloti lpsB complements a mutant of R. leguminosarum defective in lpcC, but the converse does not occur. LpcC encodes a highly selective mannosyl transferase that utilizes GDP mannose to glycosylate the inner 3-deoxy-D-manno-octulosonic acid (Kdo) residue of the lipopolysaccharide precursor Kdo(2)-lipid IV(A). We now demonstrate that LpsB can also efficiently mannosylate the same acceptor substrate as does LpcC. Unexpectedly, however, the sugar nucleotide selectivity of LpsB is greatly relaxed compared with that of LpcC. Membranes of the wild-type S. meliloti strain 2011 catalyze the glycosylation of Kdo(2)-[4'-(32)P]lipid IV(A) at comparable rates using a diverse set of sugar nucleotides, including GDP-mannose, ADP mannose, UDP-glucose, and ADP-glucose. This complex pattern of glycosylation is due entirely to LpsB, since membranes of the S. meliloti lpsB mutant 6963 do not glycosylate Kdo(2)-[4'-(32)P]lipid IV(A) in the presence of any of these sugar nucleotides. Expression of lpsB in E. coli using a T7lac promoter-driven construct results in the appearance of similar multiple glycosyl transferase activities seen in S. meliloti 2011 membranes. Constructs expressing lpcC display only mannosyl transferase activity. We conclude that LpsB, despite its high degree of similarity to LpcC, is a much more versatile glycosyltransferase, probably accounting for the inability of lpcC to complement S. meliloti lpsB mutants. Our findings have important implications for the regulation of core glycosylation in S. meliloti and other bacteria containing LpcC orthologs. PMID- 12591937 TI - A mannosyl transferase required for lipopolysaccharide inner core assembly in Rhizobium leguminosarum. Purification, substrate specificity, and expression in Salmonella waaC mutants. AB - The lipopolysaccharide (LPS) core domain of Gram-negative bacteria plays an important role in outer membrane stability and host interactions. Little is known about the biochemical properties of the glycosyltransferases that assemble the LPS core. We now report the purification and characterization of the Rhizobium leguminosarum mannosyl transferase LpcC, which adds a mannose unit to the inner 3 deoxy-d-manno-octulosonic acid (Kdo) moiety of the LPS precursor, Kdo(2)-lipid IV(A). LpcC containing an N-terminal His(6) tag was assayed using GDP-mannose as the donor and Kdo(2)-[4'-(32)P]lipid IV(A) as the acceptor and was purified to near homogeneity. Sequencing of the N terminus confirmed that the purified enzyme is the lpcC gene product. Mild acid hydrolysis of the glycolipid generated in vitro by pure LpcC showed that the mannosylation occurs on the inner Kdo residue of Kdo(2)-[4'-(32)P]lipid IV(A). A lipid acceptor substrate containing two Kdo moieties is required by LpcC, since no activity is seen with lipid IV(A) or Kdo lipid IV(A). The purified enzyme can use GDP-mannose or, to a lesser extent, ADP mannose (both of which have the alpha-anomeric configuration) for the glycosylation of Kdo(2)-[4'-(32)P]lipid IV(A). Little or no activity is seen with ADP-glucose, UDP-glucose, UDP-GlcNAc, or UDP-galactose. A Salmonella typhimurium waaC mutant, which lacks the enzyme for incorporating the inner l-glycero-d-manno heptose moiety of LPS, regains LPS with O-antigen when complemented with lpcC. An Escherichia coli heptose-less waaC-waaF deletion mutant expressing the R. leguminosarum lpcC gene likewise generates a hybrid LPS species consisting of Kdo(2)-lipid A plus a single mannose residue. Our results demonstrate that heterologous lpcC expression can be used to modify the structure of the Salmonella and E. coli LPS cores in living cells. PMID- 12591938 TI - Identification of a highly conserved domain in the androgen receptor that suppresses the DNA-binding domain-DNA interactions. AB - The androgen receptor (AR) is a ligand-regulated and sequence-specific transcription factor that activates or represses expression of target genes. Here, we show that the N terminus of AR contains an inhibitory domain located in an 81-amino acid segment lying upstream of the DNA-binding domain (DBD). The inhibitory domain interacted directly with DBD and repressed DBD binding to the androgen response element. Mutations of the conserved amino acid residues (K520E and R538E) within the inhibitory domain decreased its inhibiting ability in vitro and increased AR trans-activation in vivo. These data demonstrate the existence of a novel inhibitory domain in the N-terminal part of AR, which might play important roles in the regulation of AR trans-activation. PMID- 12591939 TI - The C-terminal domain phosphatase and transcription elongation activities of FCP1 are regulated by phosphorylation. AB - The C-terminal domain (CTD) of the largest subunit of RNA polymerase II (RNAPII) is heavily phosphorylated during the transition from transcription initiation to the establishment of an elongation-competent transcription complex. FCP1 is the only phosphatase known to be specific for the CTD of the largest subunit of RNAPII, and its activity is believed to be required to reactivate RNAPII, so that RNAPII can enter another round of transcription. We demonstrate that FCP1 is a phosphoprotein, and that phosphorylation regulates FCP1 activities. FCP1 is phosphorylated at multiple sites in vivo. The CTD phosphatase activity of phosphorylated FCP1 is stimulated by TFIIF, whereas dephosphorylated FCP1 is not. In addition to its role in the recycling of RNAPII, FCP1 also affects transcription elongation. Phosphorylated FCP1 is more active in stimulating transcription elongation than the dephosphorylated form of FCP1. We found that only phosphorylated FCP1 can physically interact with TFIIF. We set out to purify an FCP1 kinase from HeLa cells and identified casein kinase 2, which, surprisingly, displayed a negative effect on FCP1-associated activities. PMID- 12591941 TI - Molecular mechanism of recruitment of TFIIF- associating RNA polymerase C terminal domain phosphatase (FCP1) by transcription factor IIF. AB - After mRNA transcription termination in eukaryotes, the hyperphosphorylated form of RNA polymerase II (pol II0) must be recycled by TFIIF-associating C-terminal domain phosphatase (FCP1), the phosphatase responsible for dephosphorylating the C-terminal domain of the largest polymerase subunit. Transcription factor (TF) IIF stimulates the activity of FCP1, and the RNA polymerase II-associating protein 74 subunit of TFIIF forms a complex with FCP1 in both human and yeast. Here, we report a cocrystal structure of the winged-helix domain of human RNA polymerase II-associating protein 74 bound to the alpha-helical C terminus of human FCP1 (residues 944-961). These results illustrate the molecular mechanism by which TFIIF efficiently recruits FCP1 to the pol II transcription machinery for recycling of the polymerase. PMID- 12591940 TI - Polyamines protect Escherichia coli cells from the toxic effect of oxygen. AB - Wild-type Escherichia coli cells grow normally in 95% O(2)/5% CO(2). In contrast, cells that cannot make polyamines because of mutations in the biosynthetic pathway are rapidly killed by incubation in 95% O(2)/5% CO(2). Addition of polyamines prevents the toxic effect of oxygen, permitting cell survival and optimal growth. Oxygen toxicity can also be prevented if the growth medium contains an amino acid mixture or if the polyamine-deficient cells contain a manganese-superoxide dismutase (Mn-SOD) plasmid. Partial protection is afforded by the addition of 0.4 M sucrose or 0.4 M sorbitol to the growth medium. We also report that concentrations of H(2)O(2) that are nontoxic to wild-type cells or to mutant cells pretreated with polyamines kill polyamine-deficient cells. These results show that polyamines are important in protecting cells from the toxic effects of oxygen. PMID- 12591942 TI - Selective modulation of excitatory and inhibitory microcircuits by dopamine. AB - Dopamine plays an important role in the working memory functions of the prefrontal cortex, functions that are impacted in age-related memory decline, drug abuse, and a wide variety of disorders, including schizophrenia and Parkinson's disease. We have previously reported that dopamine depresses excitatory transmission between pyramidal neurons in the prefrontal cortex. Here, using paired recordings, we have investigated dopaminergic modulation of excitatory transmission from pyramidal neurons to fast-spiking (FS) interneurons. In contrast to its effect on recurrent excitation, dopamine was without effect on excitatory transmission to FS interneurons. However, dopamine has directly enhanced the excitability of the FS interneurons to the extent that even a single excitatory postsynaptic potential could initiate spiking with great temporal precision in some of them. These results indicate that dopamine's effects on excitatory transmission are target-specific and that the axon terminals of pyramidal neurons can be selectively regulated at the level of individual synapses. Thus, dopamine's net inhibitory effect on cortical function is remarkably constrained by the nature of the microcircuit elements on which it acts. PMID- 12591943 TI - Extraordinarily slow binding of guanosine to the Tetrahymena group I ribozyme: implications for RNA preorganization and function. AB - The Tetrahymena ribozyme derived from the self-splicing group I intron binds a 5' splice site analog (S) and guanosine (G), catalyzing their conversion to a 5' exon analog (P) and GA. Herein, we show that binding of guanosine is exceptionally slow, limiting the reaction at near neutral pH. Our results implicate a conformational rearrangement on guanosine binding, likely because the binding site is not prearranged in the absence of ligand. The fast accommodation of guanosine (10(2) to 10(3) x s(-1)) and prior structural data suggest local rather than global rearrangements, raising the possibility that folding of this and perhaps other large RNAs is not fully cooperative. Guanosine binding is accelerated by addition of residues that form helices, referred to as P9.0 and P10, immediately 5' and 3' to the guanosine. These rate enhancements provide evidence for binding intermediates that have the adjacent helices formed before accommodation of guanosine into its binding site. Because the ability to form the P9.0 and P10 helices distinguishes the guanosine at the correct 3'-splice site from other guanosine residues, the faster binding of the correct guanosine can enhance specificity of 3'-splice site selection. Thus, paradoxically, the absence of a preformed binding site and the resulting slow guanosine binding can contribute to splicing specificity by providing an opportunity for the adjacent helices to increase the rate of binding of the guanosine specifying the 3'-splice site. PMID- 12591944 TI - Separable features of visual cortical plasticity revealed by N-methyl-D-aspartate receptor 2A signaling. AB - How individual receptive field properties are formed in the maturing sensory neocortex remains largely unknown. The shortening of N-methyl-d-aspartate (NMDA) receptor currents by 2A subunit (NR2A) insertion has been proposed to delimit the critical period for experience-dependent refinement of circuits in visual cortex. In mice engineered to maintain prolonged NMDA responses by targeted deletion of NR2A, the sensitivity to monocular deprivation was surprisingly weakened but restricted to the typical critical period and delayed normally by dark rearing from birth. Orientation preference instead failed to mature, occluding further effects of dark rearing. Interestingly, a full ocular dominance plasticity (but not orientation bias) was selectively restored by enhanced inhibition, reflecting an imbalanced excitation in the absence of NR2A. Many of the downstream pathways involved in NMDA signaling are coupled to the receptor through a variety of protein-protein interactions and adaptor molecules. To further investigate a mechanistic dissociation of receptive field properties in the developing visual system, mice carrying a targeted disruption of the NR2A-associated 95-kDa postsynaptic density (PSD95) scaffolding protein were analyzed. Although the development and plasticity of ocular dominance was unaffected, orientation preference again failed to mature in these mice. Taken together, our results demonstrate that the cellular basis generating individual sensory response properties is separable in the developing neocortex. PMID- 12591946 TI - Mechanisms associated with cGMP binding and activation of cGMP-dependent protein kinase. AB - Using small-angle x-ray scattering, we have observed the cGMP-induced elongation of an active, cGMP-dependent, monomeric deletion mutant of cGMP-dependent protein kinase (Delta(1-52)PKG-I beta). On saturation with cGMP, the radius of gyration of Delta(1-52)PKG-I beta increases from 29.4 +/- 0.1 A to 40.1 +/- 0.7 A, and the maximum linear dimension increases from 90 A +/- 10% to 130 A +/- 10%. The elongation is due to a change in the interaction between structured regulatory (R) and catalytic (C) domains. A model of cGMP binding to Delta(1-52)PKG-I beta indicates that elongation of Delta(1-52)PKG-I beta requires binding of cGMP to the low-affinity binding site of the R domain. A comparison with cAMP-dependent protein kinase suggests that both elongation and activation require cGMP binding to both sites; cGMP binding to the low-affinity site therefore seems to be a necessary, but not sufficient, condition for both elongation and activation of Delta(1-52)PKG-I beta. We also predict that there is little or no cooperativity in cGMP binding to the two sites of Delta(1-52)PKG-I beta under the conditions used here. Results obtained by using the Delta(1-52)PKG-I beta monomer indicate that a previously observed elongation of PKG-I alpha is consistent with a pure change in the interaction between the R domain and the C domain, without alteration of the dimerization interaction. This study has revealed important features of molecular mechanisms in the biochemical network describing PKG-I beta activation by cGMP, yielding new insight into ligand activation of cyclic nucleotide-dependent protein kinases, a class of regulatory proteins that is key to many cellular processes. PMID- 12591945 TI - An agonist-induced switch in G protein coupling of the gonadotropin-releasing hormone receptor regulates pulsatile neuropeptide secretion. AB - The pulsatile secretion of gonadotropin-releasing hormone (GnRH) from normal and immortalized hypothalamic GnRH neurons is highly calcium-dependent and is stimulated by cAMP. It is also influenced by agonist activation of the endogenous GnRH receptor (GnRH-R), which couples to G(q/11) as indicated by release of membrane-bound alpha(q/11) subunits and increased inositol phosphate/Ca(2+) signaling. Conversely, GnRH antagonists increase membrane-associated alpha(q/11) subunits and abolish pulsatile GnRH secretion. GnRH also stimulates cAMP production but at high concentrations has a pertussis toxin-sensitive inhibitory effect, indicative of receptor coupling to G(i). Coupling of the agonist activated GnRH-R to both G(s) and G(i) proteins was demonstrated by the ability of nanomolar GnRH concentrations to reduce membrane-associated alpha(s) and alpha(i3) levels and of higher concentrations to diminish alpha(i3) levels. Conversely, alpha(i3) was increased during GnRH antagonist and pertussis toxin treatment, with concomitant loss of pulsatile GnRH secretion. In cholera toxin treated GnRH neurons, decreases in alpha(s) immunoreactivity and increases in cAMP production paralleled the responses to nanomolar GnRH concentrations. Treatment with cholera toxin and 8-bromo-cAMP amplified episodic GnRH pulses but did not affect their frequency. These findings suggest that an agonist concentration-dependent switch in coupling of the GnRH-R between specific G proteins modulates neuronal Ca(2+) signaling via G(s)-cAMP stimulatory and G(i) cAMP inhibitory mechanisms. Activation of G(i) may also inhibit GnRH neuronal function and episodic secretion by regulating membrane ion currents. This autocrine mechanism could serve as a timer to determine the frequency of pulsatile GnRH release by regulating Ca(2+)- and cAMP-dependent signaling and GnRH neuronal firing. PMID- 12591947 TI - Cholinergic plasticity in the hippocampus. AB - Tests were made for use-dependent plasticity in the cholinergic projections to hippocampus. Transient infusion of the cholinergic agonist carbachol into hippocampal slices induced rhythmic activity that persisted for hours after washout. Comparable effects were obtained with physostigmine, a drug that blocks acetylcholine breakdown and thereby enhances cholinergic transmission. It thus seems that activation of cholinergic synapses induces lasting changes in hippocampal physiology. Two lines of evidence indicated that cholinergic synapses are also the sites at which the plasticity is expressed. First, the induction and expression of the rhythms were not blocked by the N-methyl-D-aspartate receptor antagonist D-2-amino-5-phosphonovaleric acid, indicating that a long-term potentiation effect between pyramidal cells was not involved. Second, a muscarinic antagonist (atropine) completely abolished stable rhythmic activity after agonist washout. This result indicates that endogenous cholinergic activity is responsible for the persistence of rhythmic oscillations. These experiments suggest that short periods of intense cholinergic activity induce lasting changes in cholinergic synapses and thus extend such forms of plasticity to beyond the glutamatergic system. PMID- 12591948 TI - Launching the yeast 23S RNA Narnavirus shows 5' and 3' cis-acting signals for replication. AB - Narnavirus 23S RNA is a persistent positive-stranded RNA virus found in yeast Saccharomyces cerevisiae. The viral genome (2.9 kb) only encodes its RNA dependent RNA polymerase, p104. Here we report the generation of 23S RNA virus, with high frequency, from a vector containing the entire viral cDNA sequence. When the conserved GDD (Gly-Asp-Asp) motif of RNA-dependent RNA polymerase was modified, the vector failed to generate the virus, indicating that an active p104 is essential for replication. Successful launching required transcripts having the proper viral 3' terminus generated in vivo. This was accomplished through in vivo processing of the primary transcripts by the hepatitis delta virus antigenomic ribozyme directly fused to the 3' terminus of the 23S RNA genome. Although the primary transcripts also contained extra nucleotides at their 5' ends derived from the vector, the launched virus possessed the authentic 5' terminus of the viral genome without these extra nucleotides. Modifications of the genome sequence at the 5' and 3' termini abolished viral generation, indicating that the viral genome has cis-acting signals for replication at both termini. The great ease to generate the virus will facilitate the identification of these cis-acting signals. Furthermore, the virus, once generated, can be transmitted to daughter cells indefinitely without the vector or any selection, which makes the 23S RNA virus-launching system particularly useful for investigating the basis for RNA virus persistence. PMID- 12591949 TI - The integrity of a cholesterol-binding pocket in Niemann-Pick C2 protein is necessary to control lysosome cholesterol levels. AB - The neurodegenerative disease Niemann-Pick Type C2 (NPC2) results from mutations in the NPC2 (HE1) gene that cause abnormally high cholesterol accumulation in cells. We find that purified NPC2, a secreted soluble protein, binds cholesterol specifically with a much higher affinity (K(d) = 30-50 nM) than previously reported. Genetic and biochemical studies identified single amino acid changes that prevent both cholesterol binding and the restoration of normal cholesterol levels in mutant cells. The amino acids that affect cholesterol binding surround a hydrophobic pocket in the NPC2 protein structure, identifying a candidate sterol-binding location. On the basis of evolutionary analysis and mutagenesis, three other regions of the NPC2 protein emerged as important, including one required for efficient secretion. PMID- 12591950 TI - JNK phosphorylation of Bim-related members of the Bcl2 family induces Bax dependent apoptosis. AB - The c-Jun NH(2)-terminal kinase (JNK) is activated when cells are exposed to environmental stress, including UV radiation. Gene disruption studies demonstrate that JNK is essential for UV-stimulated apoptosis mediated by the mitochondrial pathway by a Bax/Bak-dependent mechanism. Here, we demonstrate that JNK phosphorylates two members of the BH3-only subgroup of Bcl2-related proteins (Bim and Bmf) that are normally sequestered by binding to dynein and myosin V motor complexes. Phosphorylation by JNK causes release from the motor complexes. These proapoptotic BH3-only proteins therefore provide a molecular link between the JNK signal transduction pathway and the Bax/Bak-dependent mitochondrial apoptotic machinery. PMID- 12591951 TI - Insulin resistance is a poor predictor of type 2 diabetes in individuals with no family history of disease. AB - In normoglycemic offspring of two type 2 diabetic parents, low insulin sensitivity (S(I)) and low insulin-independent glucose effectiveness (S(G)) predict the development of diabetes one to two decades later. To determine whether low S(I), low S(G,) or low acute insulin response to glucose are predictive of diabetes in a population at low genetic risk for disease, 181 normoglycemic individuals with no family history of diabetes (FH-) and 150 normoglycemic offspring of two type 2 diabetic parents (FH+) underwent i.v. glucose tolerance testing (IVGTT) between the years 1964-82. During 25 +/- 6 years follow-up, comprising 2,758 person years, the FH- cohort (54 +/- 9 years) had an age-adjusted incidence rate of type 2 diabetes of 1.8 per 1,000 person years, similar to that in other population-based studies, but significantly lower than 16.7 for the FH+ cohort. Even when the two study populations were subdivided by initial values of S(I) and S(G) derived from IVGTT's performed at study entry, there was a 10- to 20-fold difference in age-adjusted incidence rates for diabetes in the FH- vs. FH+ individuals with low S(I) and low S(G). The acute insulin response to glucose was not predictive of the development of diabetes when considered independently or when assessed as a function of S(I), i.e., the glucose disposition index. These data demonstrate that low glucose disposal rates are robustly associated with the development of diabetes in the FH+ individuals, but insulin resistance per se is not sufficient for the development of diabetes in individuals without family history of disease and strongly suggest a familial factor, not detectable in our current measures of the dynamic responses of glucose or insulin to an IVGTT is an important risk factor for type 2 diabetes. Low S(I) and low S(G), both measures of glucose disposal, interact with this putative familial factor to result in a high risk of type 2 diabetes in the FH+ individuals, but not in the FH- individuals. PMID- 12591952 TI - Infertility and aneuploidy in mice lacking a type IA DNA topoisomerase III beta. AB - We report that disruption of the mouse TOP3 beta gene encoding DNA topoisomerase III beta, one of the two mammalian type IA DNA topoisomerases, leads to a progressive reduction in fecundity. The litter size in crosses of top3 beta(-/-) mice decreases over time and through successive generations, and this decrease seems to reflect embryonic death rather than impaired fertilization. These observations are suggestive of a gradual accumulation of chromosomal defects in germ cells lacking DNA topoisomerase III beta, and this interpretation is supported by the observation of a high incidence of aneuploidy in the spermatocytes of infertile top3 beta(-/-) males. Cytogenetic examination of spermatocytes of wild-type mice also indicates that DNA topoisomerase III beta becomes prominently associated with the asynaptic regions of the XY bivalents during pachytene, and that there is a time lag between the appearance of chromosome-bound DNA topoisomerase III beta and Rad51, a protein known to be involved in an early step of homologous recombination. We interpret these findings, together with the known mechanistic characteristics of different subfamilies of DNA topoisomerases, in terms of a specific role of a type IA DNA topoisomerase in the resolution of meiotic double-Holliday junctions without crossing over. This interpretation is most likely applicable to mitotic cells as well and can explain the universal presence of at least one type IA DNA topoisomerase in all organisms. PMID- 12591953 TI - Transgene produces massive overexpression of human beta -glucuronidase in mice, lysosomal storage of enzyme, and strain-dependent tumors. AB - beta-Glucuronidase (GUSB) is a lysosomal enzyme important in the normal step-wise degradation of glycosaminoglycans. Deficiency of GUSB causes the lysosomal storage disease mucopolysaccharidosis VII (MPS VII, Sly disease). Affected patients have widespread progressive accumulation of beta-glucuronide-containing glycosaminoglycans in lysosomes. Enzyme replacement, bone marrow transplantation, and gene therapy can correct lysosomal storage in the MPS VII mouse model. Gene therapy in MPS VII patients and animals may result in massive overexpression of GUSB in individual tissues, and the toxicity of such overexpression is incompletely investigated. To gain insight into the effect of massive overexpression of GUSB, we established 19 transgenic mouse lines, two of which expressed very high levels of human GUSB in many tissues. The founder overexpressing mice had from >100- to several thousand-fold increases in tissue and serum GUSB. The enzyme expression in most tissues decreased in subsequent generations in one line, and expression in liver and marrow fell in subsequent generations of the other. Both lines had morphologically similar widespread lysosomal storage of GUSB and secondary elevations of other lysosomal enzymes, a finding characteristic of lysosomal storage disease. One line developed tumors, and one did not. These transgenic models show that massive overexpression of a lysosomal enzyme can be associated with dramatic morphological alterations, which, at least in one of the two lines, had little clinical consequence. For the other transgenic line, the high frequency of tumor development in F(2) FVB progeny suggests that the vector used to generate the transgenic lines has an integration site-dependent potential to be oncogenic, at least in this strain background. PMID- 12591954 TI - Structure of a cholesterol-binding protein deficient in Niemann-Pick type C2 disease. AB - Niemann-Pick disease type C2 (NP-C2) is a fatal hereditary disease characterized by accumulation of low-density lipoprotein-derived cholesterol in lysosomes. Here we report the 1.7-A resolution crystal structure of the cholesterol-binding protein deficient in this disease, NPC2, and the characterization of its ligand binding properties. Human NPC2 binds the cholesterol analog dehydroergosterol with submicromolar affinity at both acidic and neutral pH. NPC2 has an Ig-like fold stabilized by three disulfide bonds. The structure of the bovine protein reveals a loosely packed region penetrating from the surface into the hydrophobic core that forms adjacent small cavities with a total volume of approximately 160 A(3). We propose that this region represents the incipient cholesterol-binding site that dilates to accommodate an approximately 740-A(3) cholesterol molecule. PMID- 12591955 TI - De novo protein synthesis is required for the activation-induced cytidine deaminase function in class-switch recombination. AB - Activation-induced cytidine deaminase (AID) is required for class-switch recombination (CSR), somatic hypermutation, and gene conversion of Ig genes. Although AID has sequence similarity to an RNA-editing enzyme Apobec-1, how AID functions in CSR and somatic hypermutation is unknown. Because involvement of RNA editing but not DNA-editing in CSR requires de novo protein synthesis after AID expression, we examined whether protein synthesis inhibitors could block CSR in the presence of the AID activity. For this purpose we constructed AID fused with the hormone-binding domain of the estrogen receptor (AID-ER), which was introduced into AID-deficient spleen B cells. When such transfectants were treated with an estrogen analogue, 4-hydroxytamoxifen (OHT), CSR was induced within 1 h. Cycloheximide or puromycin drastically suppressed OHT-induced CSR in AID-ER expressing AID-/- B cells when added 1 h before OHT but not after OHT, suggesting that de novo protein synthesis is required for an event downstream to AID expression in CSR. The results lend the weight to RNA-editing hypothesis for the function of AID. PMID- 12591957 TI - Probing the kinesin reaction cycle with a 2D optical force clamp. AB - With every step it takes, the kinesin motor undergoes a mechanochemical reaction cycle that includes the hydrolysis of one ATP molecule, ADPP(i) release, plus an unknown number of additional transitions. Kinesin velocity depends on both the magnitude and the direction of the applied load. Using specialized apparatus, we subjected single kinesin molecules to forces in differing directions. Sideways and forward loads up to 8 pN exert only a weak effect, whereas comparable forces applied in the backward direction lead to stall. This strong directional bias suggests that the primary working stroke is closely aligned with the microtubule axis. Sideways loads slow the motor asymmetrically, but only at higher ATP levels, revealing the presence of additional, load-dependent transitions late in the cycle. Fluctuation analysis shows that the cycle contains at least four transitions, and confirms that hydrolysis remains tightly coupled to stepping. Together, our findings pose challenges for models of kinesin motion. PMID- 12591956 TI - Oligomeric and polymeric aggregates formed by proteins containing expanded polyglutamine. AB - Neurological diseases resulting from proteins containing expanded polyglutamine (polyQ) are characteristically associated with insoluble neuronal inclusions, usually intranuclear, and neuronal death. We describe here oligomeric and polymeric aggregates formed in cells by expanded polyQ. These aggregates are not dissociated by concentrated formic acid, an extremely effective solvent for otherwise insoluble proteins. Perinuclear inclusions formed in cultured cells by expanded polyQ can be completely dissolved in concentrated formic acid, but a soluble protein oligomer containing the expanded polyQ and released by the formic acid is not dissociated to monomer. In Huntington's disease, a formic acid resistant oligomer is present in cerebral cortex, but not in cerebellum. Cortical nuclei contain a polymeric aggregate of expanded polyQ that is insoluble in formic acid, does not enter polyacrylamide gels, but is retained on filters. This finding shows that the process of polymerization is more advanced in the cerebral cortex than in cultured cells. The resistance of oligomer and polymer to formic acid suggests the participation of covalent bonds in their stabilization. PMID- 12591958 TI - Detection of secreted peptides by using hypothesis-driven multistage mass spectrometry. AB - A method is presented for the rapid detection and characterization of trace amounts of peptides secreted from microorganisms, including pheromones, virulence factors, and quorum-sensing peptides. The procedure, based on targeted multistage MS, uses a novel matrix-assisted laser desorptionionization-ion trap mass spectrometer to overcome limitations of current MS methods (limited dynamic range, signal suppression effects, and chemical noise) that impair observation of low abundance peptides from complex biological matrixes. Here, secreted peptides that are hypothesized to be present in the supernatant, but that may not be sufficiently abundant to be observed in single-stage mass spectra, are subjected to multistage MS. Highly specific fragmentation signatures enable unambiguous identification of the peptides of interest and differentiation of the signals from the background. As examples, we demonstrate the rapid (<1 min) determination of the mating type of cells in colonies of Saccharomyces cerevisiae and the elucidation of autoinducing peptides (AIPs) from supernatants of pathogenic Staphylococci. We confirm the primary structures of the agrD encoded cyclic AIPs of Staphylococcus aureus for groups I, II, and IV and provide direct evidence that the native group-III AIP is a heptapeptide (INCDFLL). We also show that the homologous peptide from Staphylococcus intermedius is a nonapeptide (RIPTSTGFF) with a lactone ring formed through condensation of the serine side chain with the C terminus of the peptide. This is the first demonstration of cyclization in a staphylococcal AIP that occurs via lactone formation. These examples demonstrate the analytical power of the present procedure for characterizing secreted peptides and its potential utility for identifying microorganisms. PMID- 12591959 TI - Structural basis by which alternative splicing confers specificity in fibroblast growth factor receptors. AB - Binding specificity between fibroblast growth factors (FGFs) and their receptors (FGFRs) is essential for mammalian development and is regulated primarily by two alternatively spliced exons, IIIb ("b") and IIIc ("c"), that encode the second half of Ig-like domain 3 (D3) of FGFRs. FGF7 and FGF10 activate only the b isoform of FGFR2 (FGFR2b). Here, we report the crystal structure of the ligand binding portion of FGFR2b bound to FGF10. Unique contacts between divergent regions in FGF10 and two b-specific loops in D3 reveal the structural basis by which alternative splicing provides FGF10-FGFR2b specificity. Structure-based mutagenesis of FGF10 confirms the importance of the observed contacts for FGF10 biological activity. Interestingly, FGF10 binding induces a previously unobserved rotation of receptor Ig domain 2 (D2) to introduce specific contacts with FGF10. Hence, both D2 and D3 of FGFR2b contribute to the exceptional specificity between FGF10 and FGFR2b. We propose that ligand-induced conformational change in FGFRs may also play an important role in determining specificity for other FGF-FGFR complexes. PMID- 12591961 TI - Breast cancer stromal myxoid changes are associated with tumor invasion and metastasis: a central role for hyaluronan. AB - The interplay between a tumor and its environment is exemplified by the morphological changes observed in the stroma of human breast cancer. These changes are evident as stromal myxoid changes. Hyaluronan, an extracellular polysaccharide that has been implicated in invasion, is one of the major constituents of the stromal myxoid changes. This study evaluated the association of these stromal changes with axillary node status, tumor grade, and mortality. The prognostic value of the stromal myxoid changes was evaluated in patients with negative axillary nodes with 10 years of follow-up. Our results showed a high level of reproducibility of our stromal myxoid changes grading system (overall kappa = 0.68). Image analysis semiquantification showed marked correlation of a strong stromal hyaluronan signal with high-grade stromal myxoid changes. In a multiple logistic regression analysis, positive nodes were associated with stromal myxoid changes, tumor size, desmoplasia, lymphocytic infiltration, high tumor grade, tumor emboli, and multifocality. Stromal myxoid changes were also associated with young age and lymphatic embolizations (P <.001). Overall, there is a weak correlation between mortality and stromal myxoid changes (P <.01). Mortality was more evident with high stromal myxoid changes grades and tumor size >2 cm (P <.008). However Cox multivariate analysis fail to show stromal myxoid changes as an independent prognostic factor. In conclusion, stromal myxoid changes with high hyaluronan concentration are strongly associated with positive nodes, tumor grade, and lymphatic emboli, thereby identifying high-risk group and reinforcing the role of hyaluronan in invasion and metastasis. PMID- 12591960 TI - Proteins with H-bond packing defects are highly interactive with lipid bilayers: Implications for amyloidogenesis. AB - We noticed that disease-related amyloidogenic proteins and especially cellular prion proteins have the highest proportion of incompletely desolvated backbone H bonds among soluble proteins. Such bonds are vulnerable to water attack and thus represent structural weaknesses. We have measured the adsorption of proteins onto phospholipid bilayers and found a strong correlation between the extent of underwrapping of backbone H bonds in the native structure of a protein and its extent of deposition on the bilayer: the less the H bond wrapping, the higher the propensity for protein-bilayer binding. These observations support the proposition that soluble proteins with amyloidogenic propensity and membrane proteins share a pervasive building motif: the underwrapped H bonds. Whereas in membrane proteins, this motif does not signal a structural vulnerability, in soluble proteins, it is responsible for their reactivity. PMID- 12591962 TI - Overexpression of tissue inhibitor of metalloproteinases-3 in intestinal and cutaneous lesions of graft-versus-host disease. AB - Matrix metalloproteinases (MMPs) have been implicated in the pathobiology of various T-cell-mediated inflammatory disorders of the intestine and skin. Their synthetic inhibitor has been shown to prevent lethal acute graft-versus-host disease in animal models. We intended to determine the expression of MMPs 1, 3, 7, 9, 10, 12, and 19 and tissue inhibitors of metalloproteinases (TIMPs) 1 and 3 in intestinal and cutaneous lesions of patients suffering from graft-versus-host disease after bone marrow transplantation. In situ hybridizations for MMPs 1, 3, 7, 10, and 12 as well as TIMPs 1 and 3 were performed using (35)S-labeled cRNA probes on intestinal (n = 13) and cutaneous specimens (n = 9) from patients with graft-versus-host disease. Immunohistochemical stainings were carried out to localize MMP-9, MMP-19, TIMP-3, and TGF-beta1 proteins, and TUNEL staining, to detect apoptotic cells. TIMP-3 mRNA and protein were detected in cutaneous lesions in areas with vacuolar degeneration of the basal epidermal layer in all skin samples, and they colocalized with apoptotic keratinocytes and partly with staining for TGF-beta. None of the MMPs examined were overexpressed in skin lesions. Signals for MMP-1 and MMP-3 mRNA was found in 10/13 and 5/13 intestinal biopsies, respectively. In the gut, MMP-19-positive epithelial cells, particularly in the crypts, were found in 10/13 samples. Expression of MMPs 7, 9, 10, and 12 was absent or very low. TIMPs 1 and 3 were expressed by stromal cells in 12/13 and 10/13 gut samples, respectively. Whereas TIMP-1 was expressed particularly by subepithelial cells where epithelium had shed away, TIMP-3 was detected in deeper areas. We conclude that MMPs are differentially regulated in the skin and gut lesions of graft-versus-host disease. In agreement with previous data on cancer cells, TIMP-3, induced by TGF-beta1, may contribute to the apoptosis of keratinocytes in cutaneous graft-versus-host disease lesions, leading to typical histopathological changes. We also conclude that MMPs play a less important role as effector molecules in intestinal graft-versus-host disease than in celiac or inflammatory bowel disease. PMID- 12591963 TI - Histopathological features of the terminal ileum in lymphocytic and collagenous colitis: a study of 32 cases and review of literature. AB - Biopsy specimens from the terminal ileum of 32 patients with the histopathological diagnosis of lymphocytic colitis or collagenous colitis and 11 control individuals were evaluated for the presence or absence of ileal mucosal abnormalities and for the number of intraepithelial lymphocytes, assessed by immunohistochemical stains for the pan T-cell marker, CD3. We found that the mean CD3 counts in patients with lymphocytic/collagenous colitis were significantly higher than those in the control group. Seven of 14 patients with collagenous colitis and 14 of 18 patients with lymphocytic colitis revealed an increase in intraepithelial T lymphocytes when compared with the control group (P =.001). Other notable changes included ileal villous atrophy in one case of lymphocytic colitis and in three cases of collagenous colitis and epithelial damage with thickened subepithelial collagen in two cases of collagenous colitis. PMID- 12591964 TI - Lobular carcinoma in situ diagnosed by core needle biopsy: when should it be excised? AB - Core needle biopsy is the preferred technique for evaluating breast masses and abnormal mammographic findings. The frequency of detection of noninvasive lobular lesions by core needle biopsy is increasing. Historically, the diagnosis of lobular carcinoma in situ has been considered a risk factor for the development of invasive carcinoma, and treatment has consisted of careful clinical follow-up with or without chemopreventive therapeutic agents such as tamoxifen citrate. We retrospectively reviewed core needle biopsy material with the primary diagnoses of lobular carcinoma in situ, atypical lobular hyperplasia, and lobular neoplasia in conjunction with clinical and radiographic findings to make recommendations as to when excision may be merited. We searched our database for core needle biopsy cases with lobular carcinoma in situ, atypical lobular hyperplasia, and lobular neoplasia as the primary diagnosis. Microcalcifications had been sampled with a stereotactically guided, 11 G Mammotome biopsy device, and masses had been sampled with an ultrasound guided, 18 G core needle. Glass slides were reviewed and histological parameters assessed. Mammographic findings were reviewed, and clinical information was obtained from the medical record. When available, excisional biopsy material was reviewed. The 2337 breast core needle biopsies performed from January 1995 to December 2001 included 35 (1.5%) with classic lobular carcinoma in situ (14), lobular neoplasia (4), and atypical lobular hyperplasia (17) as the primary diagnosis. Twelve of these 35 cases (34%) had histological evidence of microcalcifications directly associated with the lobular carcinoma in situ, lobular neoplasia, atypical lobular hyperplasia. Radiologic review revealed 21 calcifications, 6 ultrasonographic masses, and 8 mammographic masses and/or architectural distortions. Excisional biopsy had been performed in 17 cases (49%). In six cases diagnosed as in situ on core needle biopsy, excisional biopsy revealed invasive carcinoma. All of these patients had radiographically detectable masses. Eleven cases had excisional biopsies that showed histology similar to that of the core needle biopsies. The most important predictor of invasive carcinoma on excision was a synchronous mass lesion. Lobular carcinoma in situ involving adenosis and lobular carcinoma in situ with pagetoid spread on core needle biopsies did not show a histologically more aggressive lesion on excision and, therefore, may not require additional surgery. Histologically identified calcifications were associated with lobular lesions 34% of the time; however, their presence inside an in situ lobular lesion did not portend worse pathology on re-excision and should not be a criterion for excision. Based on these findings, we recommend excisional biopsy of lobular carcinoma in situ, atypical lobular hyperplasia or lobular neoplasia only when it is associated with a synchronous mass lesion. PMID- 12591965 TI - COX-2, inflammatory secreted PLA2, and cytoplasmic PLA2 protein expression in small bowel adenocarcinomas compared with colorectal adenocarcinomas. AB - Cyclooxygenase-2 (COX-2), human synovial inflammatory secreted phospholipase A2 (sPLA2) and cytoplasmic phospholipase A2 (cPLA2) are involved in eicosanoid production and also seem to participate in colorectal tumorigenesis. As there are no data regarding these enzymes in human small bowel tumors, we wanted to determine whether they were involved in human small bowel tumorigenesis, and whether their expression was different in small bowel compared to colorectal adenocarcinomas, as suggested by animal studies. We studied their protein expression by immunohistochemistry in 25 small bowel adenocarcinomas and compared it to 48 colorectal adenocarcinomas. Seventy-six percent of the small bowel and 88% of the colorectal adenocarcinomas had a moderate or strong COX-2 expression. Sixty-eight percent of the small bowel and 67% of the colorectal adenocarcinomas had a moderate or strong sPLA2 expression. Forty-eight percent of the small bowel and 35% of the colorectal adenocarcinomas had a moderate or strong cPLA2 expression. In conclusion, the increased expression of COX-2, sPLA2, and sometimes cPLA2 in both small bowel and colorectal adenocarcinomas is in accordance with the likely eicosanoid involvement in tumor development. The same pattern of protein expression found in both types of adenocarcinoma contradicts experimental results in mice. Moreover, our results strengthen the similarities between these two types of human cancer. PMID- 12591966 TI - Hep par 1 antibody stain for the differential diagnosis of hepatocellular carcinoma: 676 tumors tested using tissue microarrays and conventional tissue sections. AB - A well-characterized positive marker for hepatocellular differentiation would be a useful tool for the diagnosis of hepatocellular carcinoma (HCC). The recently commercially available Hep Par 1 antibody (clone OCH1E5.2.10) has been reported to be a sensitive marker for HCC in paraffin embedded sections. Of non hepatocellular tumors, occasional carcinomas have been reported to stain, most frequently gastric adenocarcinomas. This study further evaluated the staining of this antibody on a large number of neoplasms using tissue microarray technology as well as conventional tissue sections. Six hundred seventy-six tumors, including 19 cases of HCC, were tested. Eighteen of 19 cases of HCC were positive, 3 showing <5% staining. Two cases negative on the array showed focal staining when whole tissue sections from the same tumors were used. 16 of 34 cases of gastric carcinomas gave positive reactions, 4 of these showed less than 5% staining. Staining of gastric carcinomas was not limited to signet ring-type carcinomas or to areas of hepatoid differentiation. Only 1 of 11 cases of cholangiocarcinoma showed focal staining. We also noted several other tumors to stain occasionally, including adrenal cortical carcinoma (3/13), yolk sac tumor (2/9), colonic adenocarcinoma (8/106), lung carcinoma (3/52), ovarian carcinoma (5/48), and endocervical adenocarcinoma (1/5). We did not observe staining in pancreatic carcinoma (11), renal cell carcinoma (36), breast carcinoma (85), melanoma (25), or mesothelioma (5). This study supports Hep Par 1 as a useful marker in the differential diagnosis of HCC, but with significant limitations. Cautious use of this antibody in a panel with other positive (alpha fetoprotein, CD10, polyclonal carcinoembryonic antigen) and negative (epithelial membrane antigen, monoclonal carcinoembryonic antigen, CD15) markers of hepatocellular differentiation may aid in the accurate diagnosis of HCC. PMID- 12591967 TI - Infection of mesothelial cells with human herpes virus 8 in human immunodeficiency virus-infected patients with Kaposi's sarcoma, Castleman's disease, and recurrent pleural effusions. AB - Recurrent pleural effusions are common complications of hospitalized patients with human immunodeficiency virus (HIV) infection and may pose difficult diagnostic dilemmas. A common cause of recurrent pleural effusions in up to 30% of HIV-seropositive patients is pulmonary involvement by Kaposi's sarcoma, a human herpesvirus 8 (HHV 8)-related neoplasm. The pathogenesis of these effusions is unclear. These recurrent effusions, although benign, have shown significant mesothelial atypia/reactive changes of uncertain etiology. We attempted to evaluate these effusions morphologically and molecularly for the presence of HHV 8, with particular attention to mesothelial cells. All recurrent pleural effusions, as defined by any effusion tapped for cytological examination on more than two occasions, in HIV-positive patients at the National Institutes of Health were examined from 1998 to the present. Cases were stratified according to patients with and without histologically confirmed HHV 8 disease manifestations. Five patients with HHV 8 diseases (four with disseminated Kaposi's sarcoma and one with Castleman's disease) were identified. As a control group, five effusions from HIV-seropositive patients without known HHV 8-related diseases were identified. Cytological examination of effusions in patients with HHV 8-related diseases demonstrated atypical/markedly reactive mesothelial cells accompanied by a polymorphous background of lymphocytes. Molecular studies for B- and T-cell clonality in microdissected whole samples showed no definitive clones in these cases. Conversely, polymerase chain reaction (PCR) studies for the HHV 8 virus was positive in these samples. PCR studies on pure populations of microdissected mesothelial cells from the HHV 8-related effusions were positive for HHV 8 sequences, whereas those from HIV patients with non-HHV 8 related diseases were negative. Immunohistochemistry for HHV 8 (monoclonal antibody to latent nuclear antigen (LNA-1; ORF-73) on cellblock material demonstrated scattered positive mesothelial cells in three of the five cases of HHV 8-associated effusions. HHV 8 has been recently implicated in the pathogenesis of Kaposi's sarcoma and primary effusion lymphoma. Mesothelial cells in recurrent pleural effusions from patients with Kaposi's sarcoma and Castleman's disease appear to be infected with HHV 8. Additional studies need to be done to define the role of mesothelial cell infection in the pathogenesis of these HHV 8-associated effusions and define the prognostic significance. PMID- 12591968 TI - Histopathologic analysis of atypical lesions in image-guided core breast biopsies. AB - Appropriate follow-up of patients with needle core breast biopsies (NCBB) showing atypical hyperplasia remains unclear because previous studies show that subsequent open biopsies in variable proportions of these patients reveal ductal carcinoma in situ (DCIS) or even invasive carcinoma, indicating significant sampling artifact. NCBB with diagnoses of atypia were morphologically classified into groups as follows: I, ALH (n = 24); II, ADH with minimal cytologic atypism (n = 90); III, atypia, other (9 columnar, 2 apocrine, 11 atypical papillary); IV, severe ADH/borderline DCIS (n = 31). Mammographic and histologic features, including the number of foci of atypia in the NCBB and the calcification span, were then correlated with presence of DCIS or invasive tumor in subsequent open excisions. Open excisional biopsies showed more severe lesions in 12% of Group I III cases (8% in Group I, 9% in Group II, and 27% in Group III), of which 15 were DCIS and one was an invasive tubular carcinoma (0.3 cm). Of the DCIS, 60% (n = 9) were < or =5 mm, and 13 of 15 (87%) were low grade. The NCBB cavity was immediately adjacent to the more severe lesions in 88% (n = 14) of cases, in keeping with sampling error. The subset showing severe ADH with borderline nuclear features in contrast was associated with a high likelihood (63%) of DCIS in follow-up excisions. NCBB with atypical papillary features also showed a high frequency of DCIS (4/11, 36%) in subsequent open excisions. Other factors associated with more severe lesions on open biopsy included the number of atypical foci in the NCBB (>4, P <.05) and the mammographic calcification span (>2.0 cm, P <.0001). Atypical lesions diagnosed in NCBB samples are radiographically and morphologically heterogeneous, accounting for the variable frequency of DCIS or invasive neoplasm identified in subsequent open excisions, which are usually focal, low grade, and a consequence of sampling artifact (i.e., adjacent to the NCBB cavity). DCIS is more likely if microcalcifications are mammographically extensive or if atypia is multifocal or is associated with borderline cytologic features. PMID- 12591969 TI - Catalyzed signal amplification for cyclin D1 detection in mantle cell lymphoma. AB - Mantle cell lymphoma is characterized by a t(11;14)(q13;q32) translocation resulting in cyclin D1 protein overexpression. Immunohistochemical detection of the latter, therefore, is a useful marker for the diagnosis of mantle cell lymphoma. Nevertheless, interpretation of results is often hampered by the weak immunoreactivity obtained with routine detection techniques. This problem can be overcome by resorting to highly sensitive catalyzed signal amplification methods based on peroxidase-catalyzed deposition of a biotinylated phenolic compound. The present study compares the results obtained with catalyzed signal amplification, labeled streptavidin biotin, and dextran polymeric conjugate (EnVision+) techniques in cyclin D1 demonstration in mantle cell lymphoma. The study was performed on formalin-fixed, paraffin-embedded archival tissue from 20 mantle cell lymphoma cases. Ten cases of small lymphocytic lymphoma and 10 instances of follicular center cell lymphoma were used as controls. Antigen retrieval was done by autoclaving under controlled pressure (2 bar) and temperature (120 degrees C) conditions. The best results were obtained after 1 minute of exposure with catalyzed signal amplification and after 6 minutes with other detection systems. Regarding cyclin D1 expression in mantle cell lymphoma cases, 17 (85%) were weakly positive and 3 (15%), moderately positive with labeled streptavidin biotin, whereas 15 (75%) were weakly positive and 5 (25%) moderately positive with EnVision+. In contrast, all 20 mantle cell lymphoma cases were strongly cyclin D1 positive with catalyzed signal amplification. No evidence of cyclin D1 immunostaining was obtained in any of the small lymphocytic lymphoma and follicular center cell lymphoma instances with any of the three methods used. In conclusion, catalyzed signal amplification methods provide a very useful tool for cyclin D1 demonstration in cases in which other immunohistochemical techniques yield inconclusive results. PMID- 12591970 TI - Florid CD4+, CD56+ T-cell infiltrate associated with Herpes simplex infection simulating nasal NK-/T-cell lymphoma. AB - We report a case of Herpes simplex virus (HSV) infection of the nasopharynx associated with a dense CD4+, CD56+ T-cell infiltrate that simulated lymphoma on clinical, histologic, and immunophenotypic grounds. Histologic examination showed a tumorlike lymphoid infiltrate with extensive necrosis. Multinucleated giant cells with "ground-glass" nuclei characteristic of HSV were observed in necrotic areas but were not prominent. Immunohistochemical studies of the lymphoid infiltrate revealed a predominance of T cells, positive for CD3, CD4, CD5, and CD56. Immunohistochemical staining with HSV antibody was focally positive in the multinucleated giant cells. Molecular studies using PCR and Southern blot were positive for HSV Type II. PCR studies for T-cell receptor gamma and immunoglobulin heavy chain gene rearrangements showed no evidence of a clonal population. In situ hybridization studies for Epstein-Barr virus (EBV) were negative. The clinical presentation of a large fungating mass, the extent of the lymphoid infiltrate, and the expression of CD56 all raised the possibility of a nasal NK/T cell lymphoma. However, the presence of HSV, lack of angioinvasion and angiodestruction, absence of EBV, and polyclonal T-cell nature of the infiltrate argued against this diagnosis. Although prior studies have not fully characterized the immunophenotypic features of the lymphocyte response to HSV in infected tissues, we postulate that the CD56+, CD4+ T-cell reaction represents a florid antiviral immune response. PMID- 12591971 TI - Current perspectives on HER2 testing: a review of national testing guidelines. AB - Knowledge of HER2 status is a prerequisite when considering a patient's eligibility for Herceptin (trastuzumab) therapy. Accurate assessment of HER2 status is essential to ensure that all patients who may benefit from Herceptin are correctly identified. There are several assays available to determine HER2 status: the most common in routine clinical practice are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Various factors can affect the results achieved with these assays, including the assay antibody/probe, the methodology and the experience of personnel. Many countries have implemented national testing guidelines in an attempt to standardize testing procedures and make results more accurate. These guidelines vary in the level of detail and the number of recommendations. This review looks at areas of consensus between the different national testing guidelines and highlights where errors may arise during the testing procedure. The key point underlined by this review is that whatever method is used to test for HER2 status, the technology must be validated first, and there must be regular internal and external quality control and quality assurance procedures. PMID- 12591973 TI - Does practice make perfect when interpreting mammography? Part II. PMID- 12591974 TI - AKT and the phosphatidylinositol 3-kinase/AKT pathway: important molecular targets for lung cancer prevention and treatment. PMID- 12591976 TI - Dosing study seen as victory for clinical trials, mathematical models. PMID- 12591977 TI - Multidrug resistance: can new drugs help chemotherapy score against cancer? PMID- 12591978 TI - U.S. falls short of top-notch health care, report says. PMID- 12591979 TI - Accelerated partial breast irradiation: more data needed, researchers say. PMID- 12591980 TI - Stat bite: Global patterns of annual deaths by cause, 2000. PMID- 12591983 TI - Health-related quality-of-life measurement in randomized clinical trials in breast cancer--taking stock. AB - Measurement of health-related quality-of-life (HRQOL) in randomized clinical trials in breast cancer has become common. In this review, we take stock of the contribution that HRQOL measurement in breast cancer clinical trials makes to clinical decision making regarding selection of optimal treatment. A series of MEDLINE searches was conducted to identify all randomized trials in breast cancer that included self-reported HRQOL or psychosocial outcomes. A total of 256 citations were identified that included HRQOL or psychosocial outcomes in breast cancer patients, and 66 of these involved randomized clinical trials of treatment. These 66 reports of breast cancer clinical trials of treatment are discussed in this review. Forty-six of the trials evaluated biomedical interventions, and 20 evaluated psychosocial interventions. Among the biomedical trials, eight trials evaluated HRQOL in primary management of breast cancer, seven trials evaluated HRQOL in adjuvant therapy of breast cancer patients, 20 trials involved metastatic breast cancer, eight trials involved symptom control/supportive care, and three trials evaluated different approaches to investigation or follow-up of breast cancer patients. Among the psychosocial trials, 13 trials evaluated HRQOL in adjuvant therapy of breast cancer patients, and their partners or spouses, six trials involved metastatic breast cancer, and one trial focused on symptom control. We found that the contribution of HRQOL measurement to clinical decision making depended on the clinical setting. In primary management of breast cancer, where medical outcomes of several treatment options are equivalent, HRQOL measurement provided added information for clinical decision making beyond that of traditional medical outcomes. In trials in the adjuvant setting, HRQOL measurement did not influence clinical decision making. In metastatic disease, HRQOL outcomes provided little information beyond that obtained from traditional medical outcomes, including toxicity. In the symptom control/supportive care setting, results of HRQOL questionnaires targeting specific symptoms (e.g., emesis) guided treatment decisions. In psychosocial intervention trials, psychosocial and/or HRQOL measurements often provided the only outcome information; therefore, selection of instruments that captured attributes likely to be altered by the intervention was essential. Until results of ongoing trials in breast cancer are available, caution is recommended in initiating new HRQOL studies unless treatment equivalency is expected, or unless the HRQOL questions target unique or specific issues that can only be addressed through patient self-report, including outcomes of psychosocial interventions. PMID- 12591984 TI - Association of volume and volume-independent factors with accuracy in screening mammogram interpretation. AB - BACKGROUND: Early detection of breast cancer is associated with the accurate reading of screening mammograms, but factors that influence reading accuracy are not well understood. We thus investigated whether reading volume and other factors were independently associated with accuracy in reading screening mammograms in a population of U.S. radiologists. METHODS: A random selection of 110 of 292 radiologists who agreed to participate, if selected, interpreted screening mammograms from 148 randomly selected women. Original index mammograms (i.e., mediolateral oblique and craniocaudal views of each breast) were used; comparison original mammograms were provided when available. Radiologist-level and facility-level factors were surveyed. Two standard metrics of screening accuracy, both based on receiver operating characteristic curves, were analyzed. The influence of volume on accuracy after controlling for other factors was assessed with multiple regression analysis. RESULTS: Current reading volume was not statistically significantly associated with interpretive accuracy. More recently trained radiologists interpreted mammograms more accurately than those trained earlier (-0.76% [95% confidence interval (CI) = -1.75% to -0.02%] reduction in sensitivity per year since residency). Facility-level factors that were statistically significantly and independently associated with better accuracy were the number of diagnostic breast imaging examinations and image guided breast interventional procedures performed (0.55% [95% CI = 0.11% to 2.40%] increase in accuracy per examination or procedure offered), being classified as a comprehensive breast diagnostic and/or screening center or freestanding mammography center (1.39% [95% CI = 0.15% to 3.82%] higher than a hospital radiology department or multispecialty medical clinic), and being a facility that practiced double reading (1.61% [95% CI = 1.99% to 11.65%]) higher than in a facility without such practice). CONCLUSIONS: Individual radiologists' current reading volume was not statistically significantly associated with accuracy in reading screening mammograms, but several other factors were. Expertise reflects a complex multifactorial process that needs further clarification. PMID- 12591985 TI - Effects of deguelin on the phosphatidylinositol 3-kinase/Akt pathway and apoptosis in premalignant human bronchial epithelial cells. AB - BACKGROUND: Because lung cancer is the leading cause of cancer-related death, new approaches for preventing and controlling the disease are needed. Chemoprevention approaches are both feasible and effective. We evaluated the potential of deguelin, a natural plant product, as a lung cancer chemopreventive agent and investigated its mechanism of action. METHODS: The effects of deguelin on proliferation and apoptosis of normal, premalignant, and malignant human bronchial epithelial (HBE) cells were assessed by using the MTT assay, a flow cytometry-based TUNEL assay, and western blot analyses. The effects of deguelin on the phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK) pathways were assessed by western blot analyses and with adenoviral vectors that expressed constitutively active Akt. RESULTS: Deguelin treatment in vitro at doses attainable in vivo inhibited the growth of and induced apoptosis of premalignant and malignant HBE cells but had minimal effects on normal HBE cells. Levels of phosphorylated Akt (pAkt) were higher in premalignant HBE cells than in normal HBE cells. In premalignant HBE cells, deguelin inhibited PI3K activity and reduced pAkt levels and activity but had mimimal effects on the MAPK pathway. Although overexpression of a constitutively active Akt in premalignant and malignant HBE cells had no effect on growth inhibition mediated by N-(4 hydroxyphenyl)retinamide (4-HPR), a novel chemopreventive retinoid, it blocked deguelin-induced growth arrest and apoptosis. CONCLUSIONS: The ability of deguelin to inhibit PI3K/Akt-mediated signaling pathways may contribute to the potency and specificity of this pro-apoptotic drug. Because both premalignant and malignant HBE cells are more sensitive to deguelin than normal HBE cells, deguelin may have potential as both a chemopreventive agent for early stages of lung carcinogenesis and a therapeutic agent against lung cancer. PMID- 12591986 TI - Effects of tamoxifen on benign breast disease in women at high risk for breast cancer. AB - BACKGROUND: In 1998 the National Surgical Adjuvant Breast and Bowel Project (NSABP) demonstrated that tamoxifen treatment reduced the incidence of both invasive and noninvasive breast cancer in women at high risk for the disease. We examined the effect of tamoxifen treatment on the incidence of benign breast disease and the number of breast biopsies in the same group of women. METHODS: We examined the medical records of 13 203 women with follow-up who participated in the NSABP Breast Cancer Prevention Trial. Included in this analysis were women who had undergone a breast biopsy and who had histologic diagnoses of adenosis, cyst, duct ectasia, fibrocystic disease, fibroadenoma, fibrosis, hyperplasia, or metaplasia. The relative risk (RR) for each histologic diagnosis was estimated for women who received tamoxifen and for women who received placebo. We also tallied the number of biopsies that women in the placebo and tamoxifen groups underwent. RESULTS: Overall, tamoxifen treatment reduced the risk of benign breast disease by 28% (RR = 0.72, 95% confidence interval [CI] = 0.65 to 0.79). Tamoxifen therapy resulted in statistically significant reductions in the risk of adenosis (RR = 0.59, 95% CI = 0.47 to 0.73), cyst (RR = 0.66, 95% CI = 0.58 to 0.75), duct ectasia (RR = 0.72, 95% CI = 0.53 to 0.97), fibrocystic disease (RR = 0.67, 95% CI = 0.58 to 0.77), hyperplasia (RR = 0.60, 95% CI = 0.50 to 0.71), and metaplasia (RR = 0.51, 95% CI = 0.41 to 0.62). Tamoxifen therapy also reduced the risk for fibroadenoma (RR = 0.77, 95% CI = 0.56 to 1.04) and fibrosis (RR = 0.86, 95% CI = 0.72 to 1.03). Compared with the placebo group, the tamoxifen group had 29% (95% CI = 23% to 34%) fewer biopsies (1048 versus 1469) and 19% fewer women who underwent a biopsy (811 versus 1019). This resulted in a 29% reduction in the risk of biopsy in women treated with tamoxifen (RR = 0.71, 95% CI = 0.66 to 0.77). This risk reduction occurred predominantly in women younger than 50 years. CONCLUSION: Women in this study who received tamoxifen, especially younger women (i.e., <50 years), had a reduced incidence of clinically detected benign breast disease and underwent fewer breast biopsies. PMID- 12591987 TI - Repair of UV light-induced DNA damage and risk of cutaneous malignant melanoma. AB - BACKGROUND: The mechanism underlying the role of UV light exposure from sunlight in the etiology of cutaneous malignant melanoma (CMM) is unclear. Patients with xeroderma pigmentosum, a disease characterized by severe sensitivity to UV radiation and a defect in nucleotide excision repair, have a high incidence of CMM, which suggests that DNA repair capacity (DRC) plays a role in sunlight induced CMM in the general population as well. METHODS: We conducted a hospital based case-control study of DRC and CMM among 312 non-Hispanic white CMM patients who had no prior chemotherapy or radiation therapy, and 324 cancer-free control subjects who were frequency-matched to case patients on age, sex, and ethnicity. Information on demographic variables, risk factors, and tumor characteristics was obtained from questionnaires and medical records. We used the host-cell reactivation assay to measure the DRC in study subjects' lymphocytes. All statistical tests were two sided. RESULTS: Case patients had a 19% lower mean (+/ standard deviation [SD]) DRC (8.5 +/- 3.4%) than control subjects (10.5 +/- 5.1%), a statistically significant difference (P<.001). DRC that was at or below the median value (i.e., 9.4%) in control subjects was associated with increased risk for CMM after adjustment for age, sex, and other covariates (odds ratio [OR] = 2.02, 95% confidence interval [CI] = 1.45 to 2.82). We observed a dose-response relationship between decreased DRC and increased risk of CMM (P(trend)<.001). Patients with tumors on sun-exposed skin had statistically significantly lower DRC than patients with tumors on unexposed skin (8.2 +/- 3.3% versus 9.5 +/- 3.5%; P =.004). CONCLUSIONS: Reduced DRC is an independent risk factor for CMM and may contribute to susceptibility to sunlight-induced CMM among the general population. PMID- 12591989 TI - Effects of 5-aza-2'-deoxycytidine on matrix metalloproteinase expression and pancreatic cancer cell invasiveness. AB - To investigate whether DNA methylation and the invasive phenotype of pancreatic adenocarcinoma are associated, we studied the role of methylation in the transcriptional regulation of several matrix metalloproteinases (MMPs) and the effect of 5-aza-2'-deoxycytidine (5Aza-dC), an inhibitor of DNA methylation, on the invasive behavior of pancreatic cancer cells. Using the Boyden chamber in vitro invasion assay, we found a statistically significant increase in invasive potential in four of five pancreatic cancer cell lines after treatment with 5Aza dC. This enhanced invasiveness was associated with the induction of mRNAs for one or more MMPs critical for tumor invasion, including MMP-1, -2, -3, -7, -9, and 14. Addition of an MMP inhibitor (GM6001, GM1489, doxycycline, or tissue inhibitor of metalloproteinase 2) blocked the 5Aza-dC-induced increase in the number of invading cells. As shown by a methylation-specific polymerase chain reaction, 5' CpG sites in MMP-2, -7, and -9 genes were partially or completely methylated in cell lines that expressed little or no corresponding mRNAs. Thus, DNA methylation influences the expression of MMP genes, and use of methylation inhibitors may stimulate the invasion of pancreatic cancer by reactivating invasion-promoting genes. PMID- 12591988 TI - Involvement of Toll-like receptor 4 signaling in interferon-gamma production and antitumor effect by streptococcal agent OK-432. AB - BACKGROUND: The streptococcal agent OK-432 has been used for immunotherapy of head and neck cancer, among other malignancies, but its mechanism of action is unknown. Because the Toll-like receptor 4 (TLR4)/MD-2 complex is important in enabling the mammalian immune system to recognize bacterial components, we investigated whether expression of the TLR4 and MD-2 genes is associated with OK 432-induced anticancer immunity. METHODS: Peripheral blood mononuclear cells (PBMCs) from 28 patients with head and neck cancer were analyzed for TLR4 and MD 2 mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR) analysis. PBMCs were treated in vitro with OK-432 or with OK-PSA (a lipoteichoic acid-related molecule that is an active component of OK-432), and interferon gamma (IFN-gamma) mRNA expression, an immune response measure, was analyzed by RT PCR. Patient sera collected 24 hours after OK-432 administration were examined for IFN-gamma protein using an enzyme-linked immunosorbent assay. Lewis lung carcinoma-bearing wild-type C57BL/6 and TLR4-deficient mice (four mice per group) received intraperitoneal injections of OK-432, and tumor volumes and sera IFN gamma levels were measured over time. All statistical tests were two-sided. RESULTS: Twenty patients expressed both TLR4 and MD-2. Expression of TLR4 and MD 2 genes was associated with the in vivo IFN-gamma induction in 19 patients administered OK-432 (Fisher's exact test P<.001). Although both OK-432 and OK-PSA induced IFN-gamma expression from PBMCs in vitro, expression of TLR4 and MD-2 was associated only with IFN-gamma expression induced by OK-PSA (P<.001). In vivo intraperitoneal administration of OK-432 resulted in an increase of IFN-gamma in sera from wild-type mice but not in sera from TLR4-deficient mice. Tumors in wild type mice treated with OK-432 were statistically significantly smaller than those in mice treated with saline (P =.007). By contrast, in TLR4-deficient mice, there was no difference in tumor volume between the two treatment groups. CONCLUSIONS: TLR4 and MD-2 may mediate OK-432-induced anticancer immunity. PMID- 12591991 TI - Re: Assessment of plasma DNA levels, allelic imbalance, and CA 125 as diagnostic tests for cancer. PMID- 12591992 TI - Re: A randomized, placebo-controlled trial of zoledronic acid in patients with hormone-refractory metastatic prostate carcinoma. PMID- 12591993 TI - Re: A randomized, placebo-controlled trial of zoledronic acid in patients with hormone-refractory metastatic prostate carcinoma. PMID- 12591994 TI - Re: A randomized, placebo-controlled trial of zoledronic acid in patients with hormone-refractory metastatic prostate carcinoma. PMID- 12591997 TI - Meeting report: in silico cell signaling underground. AB - Cell signaling is becoming a darling of systems biology. Because cell signaling is a relatively well understood, complex, but not overwhelming area of biology, it has become an attractive focal point for computational modeling and simulation efforts by engineers, computer scientists, mathematicians, and systems biologists. The flow of information within and between cells points to a possible conceptual and theoretical kinship with similar phenomena in computer networks, communication theory, information theory, and information flow in engineered systems. Although still in its infancy, the field is experiencing a growth in interest, as evidenced by the poster sessions at the recent International Conference on Systems Biology (ICSB 2002). This report focuses on some of those efforts to mathematically model and simulate cell signaling and signal transduction networks. PMID- 12591998 TI - Measuring chemotaxis and chemokinesis: the under-agarose cell migration assay. AB - Chemotaxis is the primary mechanism by which cell movements are directed within multicellular organisms, and it is a major component of embryonic development, wound healing, and immune responses. Chemotaxis involves a complex cascade of events--formation of signaling complexes, receptor polarization, adhesion molecule activation, and cytoskeletal reorganization. Previous assay methods were limited in several ways that reduced users' abilities to obtain quantitative data or to control conditions precisely. We describe a unique chemotactic assay that can incorporate multiple chemotactic gradients in different spatial and temporal combinations. In addition, this assay is easily adapted for live-cell imaging and fluorescent microscopy. With its relative simplicity, flexibility, and precision, this method is a key tool for the study of cellular chemotactic responses and the signaling processes underlying them. PMID- 12592000 TI - A uniform system for microRNA annotation. AB - MicroRNAs (miRNAs) are small noncoding RNA gene products about 22 nt long that are processed by Dicer from precursors with a characteristic hairpin secondary structure. Guidelines are presented for the identification and annotation of new miRNAs from diverse organisms, particularly so that miRNAs can be reliably distinguished from other RNAs such as small interfering RNAs. We describe specific criteria for the experimental verification of miRNAs, and conventions for naming miRNAs and miRNA genes. Finally, an online clearinghouse for miRNA gene name assignments is provided by the Rfam database of RNA families. PMID- 12591999 TI - Uridine insertion/deletion RNA editing in trypanosome mitochondria: a complex business. AB - The basic mechanism of uridine insertion/deletion RNA editing in mitochondria of kinetoplastid protists has been established for some time but the molecular details remained largely unknown. Recently, there has been significant progress in defining the molecular components of the editing reaction. A number of factors have been isolated from trypanosome mitochondria, some of which have been definitely implicated in the uridine insertion/deletion RNA editing reaction and others of which have been circumstantially implicated. Several protein complexes have been isolated which exhibit some editing activities, and the macromolecular organization of these complexes is being analyzed. In addition, there have been several important technical advances in the in vitro analysis of editing. In this review we critically examine the various factors and complexes proposed to be involved in RNA editing. PMID- 12592002 TI - Discovery and characterization of Acanthamoeba castellanii mitochondrial 5S rRNA. AB - Although 5S rRNA is a highly conserved and universal component of eubacterial, archaeal, chloroplast, and eukaryotic cytoplasmic ribosomes, a mitochondrial DNA encoded 5S rRNA has so far been identified only in land plants and certain protists. This raises the question of whether 5S rRNA is actually required for and used in mitochondrial translation. In the protist Acanthamoeba castellanii, BLAST searches fail to reveal a 5S rRNA gene in the complete mitochondrial genome sequence, nor is a 5S-sized RNA species detectable in ethidium bromide-stained gels of highly purified mitochondrial RNA preparations. Here we show that an alternative visualization technique, UV shadowing, readily detects a novel, mitochondrion-specific small RNA in A. castellanii mitochondrial RNA preparations, and that this RNA species is, in fact, a 5S rRNA encoded by the A. castellanii mitochondrial genome. These results emphasize the need for caution when interpreting negative results that suggest the absence of 5S rRNA and/or a mitochondrial DNA-encoded 5S rRNA sequence in other (particularly protist) mitochondrial systems. PMID- 12592001 TI - Recurring features of local tertiary structural elements in RNA molecules exemplified by hepatitis D virus RNA. AB - Elements of local tertiary structure in RNA molecules are important in understanding structure-function relationships. The loop E motif, first identified in several eukaryotic RNAs at functional sites which share an exceptional propensity for UV crosslinking between specific bases, was subsequently shown to have a characteristic tertiary structure. Common sequences and secondary structures have allowed other examples of the E-loop motif to be recognized in a number of RNAs at sites of protein binding or other biological function. We would like to know if more elements of local tertiary structure, in addition to the E-loop, can be identified by such common features. The highly structured circular RNA genome of the hepatitis D virus (HDV) provides an ideal test molecule because it has extensive internal structure, a UV-crosslinkable tertiary element, and specific sites for functional interactions with proteins including host PKR. We have now found a UV-crosslinkable element of local tertiary structure in antigenomic HDV RNA which, although differing from the E loop, has a very similar pattern of sequence and secondary structure to the UV crosslinkable element found in the genomic strand. Despite the fact that the two structures map close to one another, the sequences comprising them are not the templates for each other. Instead, the template regions for each element are additional sites for potential higher order structure on their respective complementary strands. This wealth of recurring sequences interspersed with base paired stems provides a context to examine other RNA species for such features and their correlations with biological function. PMID- 12592003 TI - The role of a clinically important mutation in the fold and RNA-binding properties of KH motifs. AB - We have investigated the role in the fold and RNA-binding properties of the KH modules of a hydrophobic to asparagine mutation of clinical importance in the fragile X syndrome. The mutation involves a well-conserved hydrophobic residue close to the N terminus of the second helix of the KH fold (alpha2(3) position). The effect of the mutation has been long debated: Although the mutant has been shown to disrupt the three-dimensional fold of several KH domains, the residue seems also to be directly involved in RNA binding, the main function of the KH module. Here we have used the KH3 of Nova-1, whose structure is known both in isolation and in an RNA complex, to study in detail the role of the alpha2(3) position. A detailed comparison of Nova KH3 structure with its RNA/KH complex and with other KH structures suggests a dual role for the alpha2(3) residue, which is involved both in stabilizing the hydrophobic core and in RNA contacts. We further show by nuclear magnetic resonance (NMR) studies in solution that L447 of Nova-1 in position alpha2(3) is in exchange in the absence of RNA, and becomes locked in a more rigid conformation only upon formation of an RNA complex. This implies that position alpha2(3) functions as a "gate" in the mechanism of RNA recognition of KH motifs based on the rigidification of the fold upon RNA binding. PMID- 12592004 TI - Absence of transitive and systemic pathways allows cell-specific and isoform specific RNAi in Drosophila. AB - RNA interference (RNAi) designates the multistep process by which double-stranded RNA induces the silencing of homologous endogenous genes. Some aspects of RNAi appear to be conserved throughout evolution, including the processing of trigger dsRNAs into small 21-23-bp siRNAs and their use to guide the degradation of complementary mRNAs. Two remarkable features of RNAi were uncovered in plants and Caenorhabditid elegans. First, RNA-dependent RNA polymerase activities allow the synthesis of siRNA complementary to sequences upstream of or downstream from the initial trigger region in the target mRNA, leading to a transitive RNAi with sequences that had not been initially targeted. Secondly, systemic RNAi may cause the targeting of gene silencing in one tissue to spread to other tissues. Using transgenes expressing dsRNA, we investigated whether transitive and systemic RNAi occur in Drosophila. DsRNA-producing transgenes targeted RNAi to specific regions of alternative mRNA species of one gene without transitive effect directed to sequences downstream from or upstream of the initial trigger region. Moreover, specific expression of a dsRNA, using either cell-specific GAL4 drivers or random clonal activation of a GAL4 driver, mediated a cell-autonomous RNAi. Together, our results provide evidence that transitive and systemic aspects of RNAi are not conserved in Drosophila and demonstrate that dsRNA-producing transgenes allow powerful reverse genetic approaches to be conducted in this model organism, by knocking down gene functions at the resolution of a single-cell type and of a single isoform. PMID- 12592005 TI - Coordination of editing and splicing of glutamate receptor pre-mRNA. AB - Adenosine deaminase that acts on RNA, ADAR, catalyzes the conversion of adenosine into inosine within double-stranded RNA. This type of editing has mainly been found in genes involved in neurotransmission. Site-specific A to I modifications often require intronic sequences to create the double-stranded structure necessary for editing. A system was developed to investigate if editing and splicing of pre-mRNA are coordinated. We have focused on a selectively edited site (R/G) in the glutamate receptor subunit B pre-mRNA. This editing site is situated in close proximity to a 5' splice site. To ensure efficient splicing, the editing site, together with its natural 5' splice site, was fused to a 3' splice site of the major late transcript from adenovirus. In vitro, on a premade transcript, ADAR2 editing and splicing were found to interfere with each other. The stable stem-loop required for ADAR2 editing had a negative effect on in vitro splicing, possibly by sequestering the 5' splice site. Further, RNA helicase A was shown to overcome the splicing inhibition caused by ADAR2. In vivo, allowing cotranscriptional processing, the same construct was found to efficiently edit and splice without interference, suggesting that the two RNA processing events are coordinated. PMID- 12592006 TI - Two reactions of Haloferax volcanii RNA splicing enzymes: joining of exons and circularization of introns. AB - Archaeal RNA splicing involves at least two protein enzymes, a specific endonuclease and a specific ligase. The endonuclease recognizes and cleaves within a characteristic bulge-helix-bulge (BHB) structure formed by pairing of the regions near the two exon-intron junctions, producing 2',3'-cyclic phosphate and 5'-hydroxyl termini. The ligase joins the exons and converts the cyclic phosphate into junction phosphate. The ligated product contains a seven-base hairpin loop, in which the splice junction is in between the two 3' terminal residues of the loop. Archaeal splicing endonucleases are also involved in rRNA processing, cutting within the BHB structures formed by pairing of the 5' and 3' flanking regions of the rRNAs. Large free introns derived from pre-rRNAs have been observed as stable and abundant circular RNAs in certain Crenarchaeota, a kingdom in the domain Archaea. In the present study, we show that the cells of Haloferax volcanii, a Euryarchaeote, contain circular RNAs formed by 3',5' phosphodiester linkage between the two termini of the introns derived from their pre-tRNAs. H. volcanii ligase, in vitro, can also circularize both endonuclease cleaved introns, and non-endonuclease-produced substrates. Exon joining and intron circularization are mechanistically similar ligation reactions that can occur independently. The size of the ligated hairpin loop and position of the splice junction within this loop can be changed in in vitro ligation reactions. Overall, archaeal RNA splicing seems to involve two sets of two symmetric transesterification reactions each. PMID- 12592007 TI - Structure-function analysis of the 3' stem-loop of hepatitis C virus genomic RNA and its role in viral RNA replication. AB - Previous studies indicate that the 3' terminal 46 nt of the RNA genome of hepatitis C virus (HCV) are highly conserved among different viral strains and essential for RNA replication. Here, we describe a mutational analysis of the 3' terminal hairpin (stem-loop I) that is putatively formed by this sequence and demonstrate its role in replication of the viral RNA. We show that single base substitutions within the 6-nt loop at positions adjacent to the stem abrogate replication of a subgenomic RNA, whereas substitutions in the three apical nucleotides were well tolerated without loss of replication competence. Single point mutations were also well tolerated within the middle section of the duplex, but not at the penultimate nucleotide positions near either end of the stem. However, complementary substitutions at the -19 and -28 positions (from the 3' end) restored replication competence, providing strong evidence for the existence of the structure and its involvement in RNA replication. This was confirmed by rescue of replicating RNAs from mutants containing complementary 10-nt block substitutions at the base of the stem. Each of these RNAs contained an additional U at the 3' terminus. Further experiments indicated a strong preference for U at the 3' terminal position (followed in order by C, A, and G), and a G at the -2 position. These features of stem-loop I are likely to facilitate recognition of the 3' end of the viral RNA by the viral RNA replicase. PMID- 12592008 TI - Viroid RNA systemic spread may depend on the interaction of a 71-nucleotide bulged hairpin with the host protein VirP1. AB - Viroids are noncoding circular single-stranded RNAs that are propagated systemically in plants. VirP1 is a protein from tomato, which is an excellent host for potato spindle tuber viroid (PSTVd), and it has been isolated by virtue of its specific in vitro binding to PSTVd RNA. We report on the specific in vivo interaction of VirP1 with full-length viroid RNA as well as with subfragments in the three-hybrid system. The terminal right domain (TR) of PSTVd was identified as a strong interacting partner for VirP1. A weaker partner is provided by a right-hand subfragment of hop stunt viroid (HSVd), a viroid that infects tomato poorly. We present a sequence and structural motif of the VirP1-interacting subfragments. The motif is disturbed in the replicative but nonspreading R+ mutant of the TR. According to our in vivo and in vitro binding assays, the interaction of this mutant with VirP1 is compromised. We propose that the AGG/CCUUC motif bolsters recognition of the TR by VirP1 to achieve access of the viroid to pathways that propagate endogenous RNA systemic signals in plants. Systemic trafficking has been suggested for miRNA precursors, of which the TR, as a stable bulged hairpin 71 nt long, is quite reminiscent. PMID- 12592009 TI - Common and distinctive features of GNRA tetraloops based on a GUAA tetraloop structure at 1.4 A resolution. AB - GNRA tetraloops (N is A, C, G, or U; R is A or G) are basic building blocks of RNA structure that often interact with proteins or other RNA structural elements. Understanding sequence-dependent structural variation among different GNRA tetraloops is an important step toward elucidating the molecular basis of specific GNRA tetraloop recognition by proteins and RNAs. Details of the geometry and hydration of this motif have been based on high-resolution crystallographic structures of the GRRA subset of tetraloops; less is known about the GYRA subset (Y is C or U). We report here the structure of a GUAA tetraloop determined to 1.4 A resolution to better define these details and any distinctive features of GYRA tetraloops. The tetraloop is part of a 27-nt structure that mimics the universal sarcin/ricin loop from Escherichia coli 23S ribosomal RNA in which a GUAA tetraloop replaces the conserved GAGA tetraloop. The adenosines of the GUAA tetraloop form an intermolecular contact that is a commonplace RNA tertiary interaction called an A-minor motif. This is the first structure to reveal in great detail the geometry and hydration of a GUAA tetraloop and an A-minor motif. Comparison of tetraloop structures shows a common backbone geometry for each of the eight possible tetraloop sequences and suggests a common hydration. After backbone atom superposition, equivalent bases from different tetraloops unexpectedly depart from coplanarity by as much as 48 degrees. This variation displaces the functional groups of tetraloops implicated in protein and RNA binding, providing a recognition feature. PMID- 12592010 TI - A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed in Pichia pastoris. AB - Regulated expression of proteins involved in mammalian iron metabolism is achieved in part through the interaction of the iron regulatory proteins IRP1 and IRP2 with highly conserved RNA stem-loop structures, known as iron-responsive elements (IREs), that are located within the 5' or 3' untranslated regions of regulated transcripts. As part of an effort to determine the structures of the IRP-IRE complexes using crystallographic methods, we have developed an efficient process for obtaining functionally pure IRP1 and IRP2 that relies upon the improved overexpression (>10 mg of soluble IRP per liter of culture) of each human IRP in the yeast Pichia pastoris and large-scale purification using RNA affinity chromatography. Despite the utility of RNA affinity chromatography in the isolation of RNA-binding proteins, current methods for preparing RNA affinity matrices produce columns of low capacity and limited stability. To address these limitations, we have devised a simple method for preparing stable, reusable, high capacity RNA affinity columns. This method utilizes a bifunctional linker to covalently join a 5'-amino tethered RNA with a thiol-modified Sepharose, and can be used to load 150 nmole or more of RNA per milliliter of solid support. We demonstrate here the use of an IRE affinity column in the large-scale purification of IRP1 and IRP2, and suggest that the convenience of this approach will prove attractive in the analysis of other RNA-binding proteins. PMID- 12592011 TI - Simultaneous assignment and structure determination of a membrane protein from NMR orientational restraints. AB - A solid-state NMR approach for simultaneous resonance assignment and three dimensional structure determination of a membrane protein in lipid bilayers is described. The approach is based on the scattering, hence the descriptor "shotgun," of (15)N-labeled amino acids throughout the protein sequence (and the resulting NMR spectra). The samples are obtained by protein expression in bacteria grown on media in which one type of amino acid is labeled and the others are not. Shotgun NMR short-circuits the laborious and time-consuming process of obtaining complete sequential assignments prior to the calculation of a protein structure from the NMR data by taking advantage of the orientational information inherent to the spectra of aligned proteins. As a result, it is possible to simultaneously assign resonances and measure orientational restraints for structure determination. A total of five two-dimensional (1)H/(15)N PISEMA (polarization inversion spin exchange at the magic angle) spectra, from one uniformly and four selectively (15)N-labeled samples, were sufficient to determine the structure of the membrane-bound form of the 50-residue major pVIII coat protein of fd filamentous bacteriophage. Pisa (polarity index slat angle) wheels are an essential element in the process, which starts with the simultaneous assignment of resonances and the assembly of isolated polypeptide segments, and culminates in the complete three-dimensional structure of the protein with atomic resolution. The principles are also applicable to weakly aligned proteins studied by solution NMR spectroscopy. [The structure we determined for the membrane-bound form of the Fd bacteriophage pVIII coat protein has been deposited in the Protein Data Bank as PDB file 1MZT.] PMID- 12592012 TI - Structural basis for sequential displacement of Ca(2+) by Yb(3+) in a protozoan EF-hand calcium binding protein. AB - We have studied the displacement of Ca(2+)by the trivalent lanthanide ions (Yb(3+)) in a protozoan (Entamoeba histolytica) Ca(2+)-binding protein (EhCaBP), by NMR and thermodynamics. We have demonstrated, for the first time, how one can use in a combined fashion the utility of NMR and thermodynamics to have an insight to the relative binding specificities/affinity between Ca(2+) and Yb(3+). As revealed by the titration experiments, Yb(3+) displaces Ca(2+) from the four metal binding sites present in EhCaBP in a sequential manner. The study provides a structural origin for such a sequential Ca(2+) displacement by Yb(3+) in EhCaBP. PMID- 12592013 TI - X-ray structure of the AAC(6')-Ii antibiotic resistance enzyme at 1.8 A resolution; examination of oligomeric arrangements in GNAT superfamily members. AB - The rise of antibiotic resistance as a public health concern has led to increased interest in studying the ways in which bacteria avoid the effects of antibiotics. Enzymatic inactivation by several families of enzymes has been observed to be the predominant mechanism of resistance to aminoglycoside antibiotics such as kanamycin and gentamicin. Despite the importance of acetyltransferases in bacterial resistance to aminoglycoside antibiotics, relatively little is known about their structure and mechanism. Here we report the three-dimensional atomic structure of the aminoglycoside acetyltransferase AAC(6')-Ii in complex with coenzyme A (CoA). This structure unambiguously identifies the physiologically relevant AAC(6')-Ii dimer species, and reveals that the enzyme structure is similar in the AcCoA and CoA bound forms. AAC(6')-Ii is a member of the GCN5 related N-acetyltransferase (GNAT) superfamily of acetyltransferases, a diverse group of enzymes that possess a conserved structural motif, despite low sequence homology. AAC(6')-Ii is also a member of a subset of enzymes in the GNAT superfamily that form multimeric complexes. The dimer arrangements within the multimeric GNAT superfamily members are compared, revealing that AAC(6')-Ii forms a dimer assembly that is different from that observed in the other multimeric GNAT superfamily members. This different assembly may provide insight into the evolutionary processes governing dimer formation. PMID- 12592014 TI - Solution structure of termicin, an antimicrobial peptide from the termite Pseudacanthotermes spiniger. AB - The solution structure of termicin from hemocytes of the termite Pseudacanthotermes spiniger was determined by proton two-dimensional nuclear magnetic resonance spectroscopy and molecular modeling techniques. Termicin is a cysteine-rich antifungal peptide also exhibiting a weak antibacterial activity. The global fold of termicin consists of an alpha-helical segment (Phe4-Gln14) and a two-stranded (Phe19-Asp25 and Gln28-Phe33) antiparallel beta-sheet forming a "cysteine stabilized alphabeta motif" (CSalphabeta) also found in antibacterial and antifungal defensins from insects and from plants. Interestingly, termicin shares more structural similarities with the antibacterial insect defensins and with MGD-1, a mussel defensin, than with the insect antifungal defensins such as drosomycin and heliomicin. These structural comparisons suggest that global fold alone does not explain the difference between antifungals and antibacterials. The antifungal properties of termicin may be related to its marked hydrophobicity and its amphipatic structure as compared to the antibacterial defensins. [SWISS-PROT accession number: Termicin (P82321); PDB accession number: 1MM0.] PMID- 12592015 TI - The effect of the polyproline II (PPII) conformation on the denatured state entropy. AB - Polyproline II (PPII) is reported to be a dominant conformation in the unfolded state of peptides, even when no prolines are present in the sequence. Here we use isothermal titration calorimetry (ITC) to investigate the PPII bias in the unfolded state by studying the binding of the SH3 domain of SEM-5 to variants of its putative PPII peptide ligand, Sos. The experimental system is unique in that it provides direct access to the conformational entropy change of the substituted amino acids. Results indicate that the denatured ensemble can be characterized by at least two thermodynamically distinct states, the PPII conformation and an unfolded state conforming to the previously held idea of the denatured state as a random collection of conformations determined largely by hard-sphere collision. The probability of the PPII conformation in the denatured states for Ala and Gly were found to be significant, approximately 30% and approximately 10%, respectively, resulting in a dramatic reduction in the conformational entropy of folding. PMID- 12592016 TI - A PDZ domain-based assay for measuring HIV protease activity: assay design considerations. AB - We have recently described a biochemical detection method for peptide products of enzymatic reactions based on the formation of PDZ domain*peptide ligand complexes. The product sensor is based on using masked or cryptic PDZ domain peptide ligands as enzyme substrates. Upon enzymatic processing, a PDZ-binding motif is exposed, and the product sequence bound specifically by a Eu(3+)chelate labeled GST-PDZ ([Eu(3+)]GST-PDZ). The practical applicability of this PDZ-based detection method is determined by the affinity of the PDZ domain*peptide ligand interaction, and the efficiency of the enzyme to process the masked peptide ligand. To expand the use of this PDZ-based detection strategy to a broader range of enzymatic assays, we have taken advantage of the plasticity in ligand recognition by the variety of PDZ domains found in nature. In the original work, the PDZ3 of PSD-95 was used, which preferentially recognizes the consensus sequence Ser-X-Val-COOH. Here, we show that NHERF PDZ1, which binds to the consensus sequence Thr/Ser-X-Leu-COOH, can be used to extend the flexibility in the recognition of the carboxy-terminal amino acid of the ligand, and monitor the enzymatic activity of HIV protease. The choices of detection format, for example, TRET or ALPHA, were also investigated and influenced assay design. PMID- 12592017 TI - The conserved Glu-60 residue in Thermoanaerobacter brockii alcohol dehydrogenase is not essential for catalysis. AB - Glu-60 of the zinc-dependent Thermoanaerobacter brockii alcohol dehydrogenase (TbADH) is a strictly conserved residue in all members of the alcohol dehydrogenase (ADH) family. Unlike most other ADHs, the crystal structures of TbADH and its analogs, ADH from Clostridium beijerinckii (CbADH), exhibit a unique zinc coordination environment in which this conserved residue is directly coordinated to the catalytic zinc ion in the native form of the enzymes. To explore the role of Glu-60 in TbADH catalysis, we have replaced it by alanine (E60A-TbADH) and aspartate (E60D-TbADH). Steady-state kinetic measurements show that the catalytic efficiency of these mutants is only four- and eightfold, respectively, lower than that of wild-type TbADH. We applied X-ray absorption fine-structure (EXAFS) and near-UV circular dichroism to characterize the local environment around the catalytic zinc ion in the variant enzymes in their native, cofactor-bound, and inhibited forms. We show that the catalytic zinc site in the studied complexes of the variant enzymes exhibits minor changes relative to the analogous complexes of wild-type TbADH. These moderate changes in the kinetic parameters and in the zinc ion environment imply that the Glu-60 in TbADH does not remain bound to the catalytic zinc ion during catalysis. Furthermore, our results suggest that a water molecule replaces this residue during substrate turnover. PMID- 12592018 TI - In vitro unfolding, refolding, and polymerization of human gammaD crystallin, a protein involved in cataract formation. AB - Human gammaD crystallin (HgammaD-Crys), a major protein of the human eye lens, is a primary component of cataracts. This 174-residue primarily beta-sheet protein is made up of four Greek keys separated into two domains. Mutations in the human gene sequence encoding HgammaD-Crys are implicated in early-onset cataracts in children, and the mutant protein expressed in Escherichia coli exhibits properties that reflect the in vivo pathology. We have characterized the unfolding, refolding, and competing aggregation of human wild-type HgammaD-Crys as a function of guanidinium hydrochloride (GuHCl) concentration at neutral pH and 37 degrees C, using intrinsic tryptophan fluorescence to monitor in vitro folding. Wild-type HgammaD-Crys exhibited reversible refolding above 1.0 M GuHCl. The GuHCl unfolded protein was more fluorescent than its native counterpart despite the absence of metal or ion-tryptophan interactions. Aggregation of refolding intermediates of HgammaD-Crys was observed in both equilibrium and kinetic refolding processes. The aggregation pathway competed with productive refolding at denaturant concentrations below 1.0 M GuHCl, beyond the major conformational transition region. Atomic force microscopy of samples under aggregating conditions revealed the sequential appearance of small nuclei, thin protofibrils, and fiber bundles. The HgammaD-Crys fibrous aggregate species bound bisANS appreciably, indicating the presence of exposed hydrophobic pockets. The mechanism of HgammaD-Crys aggregation may provide clues to understanding age onset cataract formation in vivo. PMID- 12592019 TI - WW domain sequence activity relationships identified using ligand recognition propensities of 42 WW domains. AB - WW domains mediate protein-protein interactions in a number of different cellular functions by recognizing proline-containing peptide sequences. We determined peptide recognition propensities for 42 WW domains using NMR spectroscopy and peptide library screens. As potential ligands, we studied both model peptides and peptides based on naturally occurring sequences, including phosphorylated residues. Thirty-two WW domains were classified into six groups according to detected ligand recognition preferences for binding the motifs PPx(Y/poY), (p/phi)P(p,g)PPpR, (p/phi)PPRgpPp, PPLPp, (p/xi)PPPPP, and (poS/poT)P (motifs according to modified Seefeld Convention 2001). In addition to these distinct binding motifs, group-specific WW domain consensus sequences were identified. For PPxY-recognizing domains, phospho-tyrosine binding was also observed. Based on the sequences of the PPx(Y/poY)-specific group, a profile hidden Markov model was calculated and used to predict PPx(Y/poY)-recognition activity for WW domains, which were not assayed. PPx(Y/poY)-binding was found to be a common property of NEDD4-like ubiquitin ligases. PMID- 12592020 TI - Probing the structure of falcipain-3, a cysteine protease from Plasmodium falciparum: comparative protein modeling and docking studies. AB - Increasing resistance of malaria parasites to conventional antimalarial drugs is an important factor contributing to the persistence of the disease as a major health threat. The ongoing search for novel targets has resulted in identification and expression of several enzymes including cysteine proteases that are implicated in hemoglobin degradation. Falcipain-2 and falcipain-3 are considered to be the two principal cysteine proteases in this degradation, and hence, are potential drug targets. A homology model of falcipain-3 was built and validated by various structure/geometry verification tools as well as docking studies of known substrates. The correlation coefficient of 0.975 between interaction energies and K(m) values of these substrates provided additional support for the model. On comparison with the previously reported falcipain-2 homology model, the currently constructed falcipain-3 structure showed important differences between the S2 pockets that might explain the variations in the K(m) values of various substrates for these enzymes. Further, docking studies also provided insight into possible binding modes and interactions of ligands with falcipain-3. Results of the current study could be employed in de novo drug design leading to development of new antimalarial agents. PMID- 12592021 TI - Backbone dynamics of the human MIA protein studied by (15)N NMR relaxation: implications for extended interactions of SH3 domains. AB - The melanoma inhibitory activity (MIA) protein is a clinically valuable marker in patients with malignant melanoma as enhanced values diagnose metastatic melanoma stages III and IV. Here, we report the backbone dynamics of human MIA studied by (15)N NMR relaxation experiments. The folded core of human MIA is found to be rigid, but several loops connecting beta-sheets, such as the RT-loop for example, display increased mobility on picosecond to nanosecond time scales. One of the most important dynamic features is the pronounced flexibility of the distal loop, comprising residues Asp 68 to Ala 75, where motions on time scales up to milliseconds occur. Further, significant exchange contributions are observed for residues of the canonical binding site of SH3 domains including the RT-loop, the n-Src loop, for the loop comprising residues 13 to 19, which we refer to as the"disulfide loop", in part for the distal loop, and the carboxyl terminus of human MIA. The functional importance of this dynamic behavior is discussed with respect to the biological activity of several point mutations of human MIA. The results of this study suggest that the MIA protein and the recently identified highly homologous fibrocyte-derived protein (FDP)/MIA-like (MIAL) constitute a new family of secreted proteins that adopt an SH3 domain-like fold in solution with expanded ligand interactions. PMID- 12592023 TI - Asparagine 285 plays a key role in transition state stabilization in rabbit muscle creatine kinase. AB - To explore the possibility that asparagine 285 plays a key role in transition state stabilization in phosphagen kinase catalysis, the N285Q, N285D, and N285A site-directed mutants of recombinant rabbit muscle creatine kinase (rmCK) were prepared and characterized. Kinetic analysis of phosphocreatine formation showed that the catalytic efficiency of each N285 mutant was reduced by approximately four orders of magnitude, with the major cause of activity loss being a reduction in k(cat) in comparison to the recombinant native CK. The data for N285Q still fit a random-order, rapid-equilibrium mechanism, with either MgATP or creatine binding first with affinities very nearly equal to those for native CK. However, the affinity for the binding of the second substrate is reduced approximately 10 fold, suggesting that addition of a single methylene group at position 285 disrupts the symphony of substrate binding. The data for the N285A mutant only fit an ordered binding mechanism, with MgATP binding first. Isosteric replacement to form the N285D mutant has almost no effect on the K(M) values for either creatine or MgATP, thus the decrease in activity is due almost entirely to a 5000 fold reduction in k(cat). Using the quenching of the intrinsic CK tryptophan fluorescence by added MgADP (Borders et al. 2002), it was found that, unlike native CK, none of the mutants have the ability to form a quaternary TSAC. We use these data to propose that asparagine 285 indeed plays a key role in transition state stabilization in the reaction catalyzed by creatine kinase and other phosphagen kinases. PMID- 12592022 TI - The hydration of amides in helices; a comprehensive picture from molecular dynamics, IR, and NMR. AB - We examined the hydration of amides of alpha(3)D, a simple, designed three-helix bundle protein. Molecular dynamics calculations show that the amide carbonyls on the surface of the protein tilt away from the helical axis to interact with solvent water, resulting in a lengthening of the hydrogen bonds on this face of the helix. Water molecules are bonded to these carbonyl groups with partial occupancy ( approximately 50%-70%), and their interaction geometries show a large variation in their hydrogen bond lengths and angles on the nsec time scale. This heterogeneity is reflected in the carbonyl stretching vibration (amide I' band) of a group of surface Ala residues. The surface-exposed amides are broad, and shift to lower frequency (reflecting strengthening of the hydrogen bonds) as the temperature is decreased. By contrast, the amide I' bands of the buried (13)C labeled Leu residues are significantly sharper and their frequencies are consistent with the formation of strong hydrogen bonds, independent of temperature. The rates of hydrogen-deuterium exchange and the proton NMR chemical shifts of the helical amide groups also depend on environment. The partial occupancy of the hydration sites on the surface of helices suggests that the interaction is relatively weak, on the order of thermal energy at room temperature. One unexpected feature that emerged from the dynamics calculations was that a Thr side chain subtly disrupted the helical geometry 4-7 residues N terminal in sequence, which was reflected in the proton chemical shifts and the rates of amide proton exchange for several amides that engage in a mixed 3(10)/alpha/pi-helical conformation. PMID- 12592024 TI - Interplay between hydrophobic cluster and loop propensity in beta-hairpin formation: a mechanistic study. AB - We investigated the structural determinants of the stability of a designed beta hairpin containing a natural hydrophobic cluster from the protein GB1 and a D-Pro Gly turn forming sequence. The results of our simulations shed light on the factors leading to an ordered secondary structure in a model peptide: in particular, the importance of the so-called diagonal interactions in forming a stable hydrophobic nucleus in the beta-hairpin, together with the more obvious lateral interactions, is examined. With the use of long timescale MD simulations in explicit water, we show the role of diagonal interactions in driving the peptide to the correct folded structure (formation of the hydrophobic core with Trp 2, Tyr 4, and Phe 9 in the first stages of refolding) and in keeping it in the ensemble of folded conformations. The combination of the stabilizing effects of the D-Pro-Gly turn sequence and of the hydrophobic nucleus formation thus favors the attainment of an ordered secondary structure compatible with the one determined experimentally. Moreover, our data underline the importance of the juxtapositions of the side chains of amino acids not directly facing each other in the three-dimensional structure. The combination of these interactions forces the peptide to sample a nonrandom portion of the conformational space, as can be seen in the rapid collapse to an ordered structure in the refolding simulation, and shows that the unfolded state can be closely correlated to the folded ensemble of structures, at least in the case of small model peptides. PMID- 12592025 TI - Purification of correctly oxidized MHC class I heavy-chain molecules under denaturing conditions: a novel strategy exploiting disulfide assisted protein folding. AB - The aim of this study has been to develop a strategy for purifying correctly oxidized denatured major histocompability complex class I (MHC-I) heavy-chain molecules, which on dilution, fold efficiently and become functional. Expression of heavy-chain molecules in bacteria results in the formation of insoluble cellular inclusion bodies, which must be solubilized under denaturing conditions. Their subsequent purification and refolding is complicated by the fact that (1). correct folding can only take place in combined presence of beta(2)-microglobulin and a binding peptide; and (2). optimal in vitro conditions for disulfide bond formation ( approximately pH 8) and peptide binding ( approximately pH 6.6) are far from complementary. Here we present a two-step strategy, which relies on uncoupling the events of disulfide bond formation and peptide binding. In the first phase, heavy-chain molecules with correct disulfide bonding are formed under non-reducing denaturing conditions and separated from scrambled disulfide bond forms by hydrophobic interaction chromatography. In the second step, rapid refolding of the oxidized heavy chains is afforded by disulfide bond-assisted folding in the presence of beta(2)-microglobulin and a specific peptide. Under conditions optimized for peptide binding, refolding and simultaneous peptide binding of the correctly oxidized heavy chain was much more efficient than that of the fully reduced molecule. PMID- 12592026 TI - Reinvestigation of the proposed folding and self-association of the Neuropeptide Head Activator. AB - The Neuropeptide Head Activator (HA), pGlu-Pro-Pro-Gly-Gly-Ser-Lys-Val-Ile-Leu Phe (pGlu is pyroglutamic acid), is involved in head-specific growth and differentiation processes in the freshwater coelenterate Hydra attenuata. Peptides of identical sequence have also been isolated from higher-organism tissues such as human and bovine hypothalamus. Early studies by molecular sieve chromatography suggested that HA dimerizes with high affinity (K(d) approximately 1 nM). This dimerization was proposed to occur via antiparallel beta-sheet formation between the Lys(7)-Phe(11) segments in each HA molecule. We conducted biophysical studies on synthetic HA in order to gain insight into its structure and aggregation tendencies. We found by analytical ultracentrifugation that HA is monomeric at low millimolar concentrations. Studies by (1)H-NMR revealed that HA did not adopt any significant secondary structure in solution. We found no NOEs that would support the proposed dimer structure. We probed the propensity of the Lys(7)-Phe(11) fragment to form antiparallel beta-sheet by designing peptides in which two such fragments are joined by a two-residue linker. These peptides were intended to form stable beta-hairpin structures with cross-strand interactions that mimic those of the proposed HA dimer interface. We found that the HA-derived fragments may be induced to form intramolecular beta-sheet, albeit only weakly, when linked by the highly beta-hairpin-promoting D-Pro-Gly turn, but not when linked by the more flexible Gly-Gly unit. These findings suggest that the postulated mode of HA dimerization and the proposed propensity of the molecule to form discrete aggregates with high affinity are incorrect. PMID- 12592027 TI - Mutations in the B1 domain of protein G that delay the onset of amyloid fibril formation in vitro. AB - We previously reported that under certain experimental conditions, many variants of the B1 domain of IgG-binding protein G from Streptococcus form fibrils reproducibly. The variant I6T53 was the focus of the present study because the lag phase in the kinetics of fibril formation by this variant is significantly longer than that of other variants. This lag phase is distinguished by changes in both intrinsic fluorescence intensity and in light scattering of the protein. NMR diffusion measurements suggest that the soluble protein during the lag phase is monomeric. The kinetic profiles of fibril formation are found to depend on experimental conditions. The first kinetic phase diminishes almost completely when the reaction is seeded with preformed amyloid fibrils. PMID- 12592028 TI - Crystal structure of a defective folding protein. AB - Maltose-binding protein (MBP or MalE) of Escherichia coli is the periplasmic receptor of the maltose transport system. MalE31, a defective folding mutant of MalE carrying sequence changes Gly 32-->Asp and Ile 33-->Pro, is either degraded or forms inclusion bodies following its export to the periplasmic compartment. We have shown previously that overexpression of FkpA, a heat-shock periplasmic peptidyl-prolyl isomerase with chaperone activity, suppresses MalE31 misfolding. Here, we have exploited this property to characterize the maltose transport activity of MalE31 in whole cells. MalE31 displays defective transport behavior, even though it retains maltose-binding activity comparable with that of the wild type protein. Because the mutated residues are in a region on the surface of MalE not identified previously as important for maltose transport, we have solved the crystal structure of MalE31 in the maltose-bound state in order to characterize the effects of these changes. The structure was determined by molecular replacement methods and refined to 1.85 A resolution. The conformation of MalE31 closely resembles that of wild-type MalE, with very small displacements of the mutated residues located in the loop connecting the first alpha-helix to the first beta-strand. The structural and functional characterization provides experimental evidence that MalE31 can attain a wild-type folded conformation, and suggest that the mutated sites are probably involved in the interactions with the membrane components of the maltose transport system. PMID- 12592029 TI - Hydrophobicity of transmembrane proteins: spatially profiling the distribution. AB - A hallmark of soluble globular protein tertiary structure is a hydrophobic core and a protein exterior populated predominantly by hydrophilic residues. Recent hydrophobic moment profiling of the spatial distribution of 30 globular proteins of diverse size and structure had revealed features of this distribution that were comparable. Analogous profiling of the hydrophobicity distribution of the alpha-helical buried bundles of several transmembrane proteins, as the lipid/protein interface is approached from within the bilayer, reveals spatial hydrophobicity profiles that contrast with those obtained for the soluble proteins. The calculations, which enable relative changes of hydrophobicity to be simply identified over the entire spatial extent of the multimer within the lipid bilayer, show the accumulated zero-order moments of the bundles to be mainly inverted with respect to that found for the soluble proteins. This indicates a statistical increase in the average residue hydrophobic content as the lipid bilayer is approached. This result differs from that of a relatively recent calculation and qualitatively agrees with earlier calculations involving lipid exposed and buried residues of the alpha-helices of transmembrane proteins. Spatial profiling, over the entire spatial extent of the multimer with scaled values of residue hydrophobicity, provides information that is not available from calculations using lipid exposure alone. PMID- 12592030 TI - Competing intrachain interactions regulate the formation of beta-sheet fibrils in bovine PrP peptides. AB - At the heart of the pathogenesis of transmissible spongiform encephalopathies (TSEs), such as BSE, scrapie, and Creutzfeldt-Jakob disease, lies a poorly understood structural rearrangement of PrP, an abundant glycoprotein of the nervous and lymphoid systems. The normal form (PrP(C)), rich in alpha-helix, converts into an aberrant beta-sheet-dominated form (PrP(Sc)), which seems to be at the center of the pathotoxic symptoms observed in TSEs. To understand this process better at a molecular level, we have studied the interactions between different peptides derived from bovine PrP and their structural significance. We show that two unstructured peptides derived from the central region of bovine PrP, residues 115-133 and 140-152, respectively, interact stoichiometrically under physiological conditions to generate beta-sheet-dominated fibrils. However, when both peptides are incubated in the presence of a third peptide derived from an adjoining alpha-helical region (residues 153-169), the formation of beta-sheet rich fibrils is abolished. These data indicate that native PrP(C) helix 1 might inhibit the strong intrinsic beta-sheet-forming propensity of sequences immediately N-terminal to the globular core of PrP(C), by keeping in place intrachain interactions that would prevent these amyloidogenic regions from triggering aggregation. Moreover, these results indicate new ways in which PrP(Sc) formation could be prevented. PMID- 12592031 TI - Equilibrium and kinetic studies on folding of canine milk lysozyme. AB - Thermal and chemical unfolding studies of the calcium-binding canine lysozyme (CL) by fluorescence and circular dichroism spectroscopy show that, upon unfolding in the absence of calcium ions, a very stable equilibrium intermediate state is formed. At room temperature and pH 7.5, for example, a stable molten globule state is attained in 3 M GdnHCl. The existence of such a pure and stable intermediate state allowed us to extend classical stopped-flow fluorescence measurements that describe the transition from the native to the unfolded form, with kinetic experiments that monitor separately the transition from the unfolded to the intermediate state and from the intermediate to the native state, respectively. The overall refolding kinetics of apo-canine lysozyme are characterized by a significant drop in the fluorescence intensity during the dead time, followed by a monoexponential increase of the fluorescence with k = 3.6 s( 1). Furthermore, the results show that, unlike its drastic effect on the stability, Ca(2+)-binding only marginally affects the refolding kinetics. During the refolding process of apo-CL non-native interactions, comparable to those observed in hen egg white lysozyme, are revealed by a substantial quenching of tryptophan fluorescence. The dissection of the refolding process in two distinct steps shows that these non-native interactions only occur in the final stage of the refolding process in which the two domains match to form the native conformation. PMID- 12592033 TI - Prediction of beta-turns in proteins from multiple alignment using neural network. AB - A neural network-based method has been developed for the prediction of beta-turns in proteins by using multiple sequence alignment. Two feed-forward back propagation networks with a single hidden layer are used where the first-sequence structure network is trained with the multiple sequence alignment in the form of PSI-BLAST-generated position-specific scoring matrices. The initial predictions from the first network and PSIPRED-predicted secondary structure are used as input to the second structure-structure network to refine the predictions obtained from the first net. A significant improvement in prediction accuracy has been achieved by using evolutionary information contained in the multiple sequence alignment. The final network yields an overall prediction accuracy of 75.5% when tested by sevenfold cross-validation on a set of 426 nonhomologous protein chains. The corresponding Q(pred), Q(obs), and Matthews correlation coefficient values are 49.8%, 72.3%, and 0.43, respectively, and are the best among all the previously published beta-turn prediction methods. The Web server BetaTPred2 (http://www.imtech.res.in/raghava/betatpred2/) has been developed based on this approach. PMID- 12592032 TI - Protein folding coupled to DNA binding in the catalytic domain of bacteriophage lambda integrase detected by mass spectrometry. AB - Bacteriophage lambda integrase (lambda-Int) is the prototypical member of a large family of enzymes that catalyze site-specific DNA recombination via single-strand cleavage and the formation of a Holliday junction intermediate. Crystallographic and biochemical evidence indicate that substantial conformational change (i.e., folding) in the catalytic domain of the protein is required for substrate recognition and catalysis. We have examined the solution conformation of the catalytic domain (C170) in the absence and presence of a cognate "half-site" DNA oligonucleotide by electrospray ionization mass spectrometry, and circular dichroism and fluorescence spectroscopy. The distribution of ions in the positive ion electrospray mass spectrum of the free protein reveals the presence of three distinct species in solution, one corresponding to the folded protein, one to the unfolded protein, and one to a dimer. In the presence of DNA, ions are observed only for the protein-DNA complex and the folded form of the free protein. We therefore conclude that DNA binding stabilizes the global fold of the protein in a manner that is consistent with folding-coupled target recognition as a mechanism to control site-specific recombination. Furthermore, we find that inspection of the charge state distribution of ions in electrospray mass spectra provides a quick and effective means to identify conformational heterogeneity of proteins in solution and to investigate dynamic protein-nucleic acid interactions. PMID- 12592034 TI - Enhanced correction methods for hydrogen exchange-mass spectrometric studies of amyloid fibrils. AB - We describe methods for minimization of and correction for artifactual forward and backward exchange occurring during hydrogen exchange-mass spectrometric (HX MS) studies of amyloid fibrils of the Abeta(1-40) peptide. The quality of the corrected data obtained using published and new correction algorithms is evaluated quantitatively. Using the new correction methods, we have determined that 20.1 +/- 1.4 of the 39 backbone amide hydrogens in Abeta(1-40) exchange with deuteriums in 100 h when amyloid fibrils of this peptide are suspended in D(2)O. These data reinforce our previous conclusions based on uncorrected data that amyloid fibrils contain a rigid protective core structure that involves only about half of the Abeta backbone amides. The methods developed here should be of general value for HX-MS studies of amyloid fibrils and other protein aggregates. PMID- 12592036 TI - Erectile dysfunction in the elderly man. PMID- 12592035 TI - Isolation and characterization of Staufen-containing ribonucleoprotein particles from rat brain. AB - Localized mRNAs are thought to be transported in defined particles to their final destination. These particles represent large protein complexes that may be involved in recognizing, transporting, and anchoring localized messages. Few components of these ribonucleoparticles, however, have been identified yet. We chose the strategy to biochemically enrich native RNA-protein complexes involved in RNA transport to identify the associated RNAs and proteins. Because Staufen proteins were implicated in intracellular RNA transport, we chose mammalian Staufen proteins as markers for the purification of RNA transport particles. Here, we present evidence that Staufen proteins exist in two different complexes: (i) distinct large, ribosome- and endoplasmic reticulum-containing granules preferentially found in the membrane pellets during differential centrifugation and (ii) smaller particles in the S100 from rat brain homogenates. On gel filtration of the S100, we identified soluble 670-kDa Staufen1-containing and 440 kDa Staufen2-containing particles. They do not cofractionate with ribosomes and endoplasmic reticulum but rather coenrich with kinesin heavy chain. Furthermore, the fractions containing the Staufen1 particles show a 15-fold enrichment of mRNAs compared with control fractions. Most importantly, these fractions are highly enriched in BC1, and, to a lesser extent, in the alpha-subunit of the Ca(2+)/calmodulin-dependent kinase II, two dendritically localized RNAs. Finally, both RNAs colocalize with Staufen1-hemagglutinin in particles in dendrites of transfected hippocampal neurons. We therefore propose that these Staufen1 containing particles may represent RNA transport intermediates that are in transit to their final destination within neurons. PMID- 12592038 TI - Tubulovasostomy. PMID- 12592037 TI - Vasovasostomy. AB - Vasovasostomy after sterilisation vasectomy is the most frequently performed microsurgical intervention in refertilizing surgery. Causes for obstruction other than vasectomy such as juvenile hernia repair and other kinds of surgery affecting the seminal cord appear to be rare. The pre-operative diagnostic approach follows the rules of conventional diagnostics in male infertility. The significance of the intra-operative evaluation of aspirates from the duct and its role in surgical decision-making is controversely debated in the literature. Similar controversies are evident concerning the microsurgical technique that is applied. In particular, the advantages of the two-layered technique (gold standard) of the anastomosis versus the single-layered technique are discussed. Other techniques such as laser applications and fibrin glue are not routinely used in humans. Even difficult microsurgical problems in cases with extended obstructions and cases of re-vasovasostomy can nowadays be managed and have significant success rates. Important prognostic factors appear to be the interval of obstruction, sperm granulomas, and length of the testicular end of the duct. High success rates accompanied by a lower profile in cost make the microsurgical approach (refertilization) superior versus the primary use of ART techniques such as ICSI. PMID- 12592039 TI - Percutaneous treatment of varicocele. AB - Varicocele is a common finding in adolescents and adult men. Its association with male infertility has been well documented: varicoceles are reported to be present in 20-40% of infertile men. It has been demonstrated that varicocele correction leads to an improvement in the quality of semen in most cases. Percutaneous sclerotherapy is an established treatment method for varicocele performed on an outpatient basis. In our report we review our experience with venographic study and transcatheter sclerotherapy based on 560 cases of infertile patients with varicocele. Our study confirms that percutaneous therapy of varicocele may lead to improved spermatogenesis in the majority of patients. PMID- 12592040 TI - Sperm retrieval procedures and intracytoplasmatic spermatozoa injection with epididymal and testicular sperms. AB - INTRODUCTION: Male infertility caused by azoospermia due to non-reconstructable obstruction or non-obstructive azoospermia can be treated by microsurgical epididymal aspiration (MESA) or testicular sperm extraction (TESE) followed by an intracytoplasmatic spermatozoa injection (ICSI). MATERIAL AND METHODS: From 9/93 to 6/01, we carried out 1,025 ICSI procedures with aspirated epididymal or testicular sperms in 684 cases. 163 ICSI cycles were performed with epididymal sperms and 862 ICSI cycles with testicular sperms or spermatids. The TESE was carried out by open biopsy, frequently in a multilocular technique. The aspirated spermatozoas were used after cryopreservation (frozen) or immediately after aspiration (fresh). RESULTS: 538 patients had obstructive azoospermia or ejaculation failure. In 487 cases the underlying cause of azoospermia was an impaired spermatogenesis, following maldescensus testis, chemotherapy, radiotherapy, or caused by Sertoli-cell-only syndrome, a genetic disorder or an unknown etiology. The transfer rates, pregnancy rates and birth rates per ICSI cycle showed no statistically significant differences between testicular and epididymal sperms in the cases of seminal obstruction (28% average birth rates in both cases). However, highly significant was the difference in birth rates with regard to the underlying cause of infertility. In contrast, in treating non obstructive azoospermia we observed a birth rate of 19% per cycle. In all patient groups the birth rate with fresh spermatozoas did not differ from those with cryopreserved spermatozoa. 40% of patients after multilocular TESE showed clinical signs of testicular lesion. CONCLUSION: The underlying cause of azoospermia is the most important factor for the outcome of ICSI using epididymal and testicular sperms. In cases of non-obstructive azoospermia, the pregnancy rate is low compared with the results in cases of obstructive azoospermia. There is no difference between fresh and cryopreserved sperms. TESE with ICSI is the most efficient treatment of azoospermia caused by hypergonadotropic hypogonadism. The morbidity of the TESE procedure is highly relevant and must be considered if this technique is indicated. PMID- 12592041 TI - Development of surgical procedures in the treatment of erectile dysfunction. A historical overview. PMID- 12592042 TI - Penile revascularization surgery in erectile dysfunction. AB - Anastomoses between the dorsal artery of penis and the deep penile artery are necessary for revascularization of penile arteries. Radiologic and anatomic proof is provided. Arterio-arterial anastomosis represents a risk factor for thrombosis. For its exclusion the necessary operative course of action is showed. By building an additional arteriovenous shunt close to the arterio-arterial anastomosis the risk of thrombosis is markedly decreased. Our technique produces good results in patients with diagnosed erectile dysfunction of vascular origin with a spontaneous erection when the occasion arises. PMID- 12592043 TI - Contemporary aspects of penile prosthesis implantation. AB - PURPOSE: Erectile dysfunction today has a number of effective treatment options. This review was undertaken to examine the contemporary role of penile prosthesis implantation in the treatment of this disorder. MATERIALS AND METHODS: A MEDLINE search was performed on the topic of penile prostheses and implants. Current literature was reviewed with regard to types of penile implants, issues related to prosthesis implantation, results, and patient/partner satisfaction. RESULTS: Mechanical failure rates for early penile prostheses, especially the inflatable type, were unacceptably high. Advances in both prosthesis design and implantation techniques have resulted in increased device survival with 5-year actuarial survival rates free of mechanical failure ranging from 86.2 to 93.6%. Recent reviews of implant recipients show 83 and 85% satisfaction and for partners 70 and 76% satisfaction. CONCLUSIONS: When systemic therapy for erectile dysfunction fails, men have a variety of other options to choose from. Penile prosthesis implantation is an option that is feasible for nearly every man with this disorder. Current device survival rates and patient and partner satisfaction rates are high. PMID- 12592044 TI - Topical nonsteroidal anti-inflammatory therapy in ophthalmology. AB - Topically applied nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used in the management and prevention of ocular inflammation and cystoid macular edema related to cataract surgery and the maintenance of mydriasis during cataract surgery. Other common uses are the reduction of discomfort after refractive surgery or in allergic conjunctivitis. NSAIDs primarily act as cyclooxygenase inhibitors and thus reduce the formation of endogenous PGs. Today, several NSAIDs are commercially available: diclofenac, flurbiprofen, indomethacin, ketorolac and suprofen. At present the ophthalmologist has to make a decision between the use of topical corticosteroids, with their potential adverse effects, or of topical NSAIDs, with their possibly increased benefit, unknown effect on ocular pressure, wound healing and corneal tissue, higher costs and limited track record. However, the improvement of surgical techniques might support an increasing use of NSAIDs in the future. Preoperative anti-inflammatory treatment should be considered in eyes at a higher risk of developing severe postoperative inflammatory reactions. This decision has to be made carefully and has to be guided by the clinical circumstances, the spectrum of diagnosis and the individual benefit-risk ratio of each patient. PMID- 12592045 TI - Comparison of endothelial cell count using confocal and contact specular microscopy. AB - Precise examination of the corneal endothelium has become increasingly important due to the growing number of intraocular and corneal procedures. The purpose of this study was to compare prospectively the corneal endothelial cell count in normal eyes obtained by confocal and specular microscopy. Central corneal endothelial cell densities of 42 eyes from 42 patients were measured by confocal and contact specular microscopy. Endothelial cells were analyzed with the same software in a manual, an automated and a semi-automated mode. The mean endothelial cell density obtained by confocal microscopy was (in the manual, automated and semi-automated modes) 3,069 +/- 285, 2,791 +/- 344 and 3,077 +/- 286 cells/mm(2), and obtained by specular microscopy 3,076 +/- 298, 2,796 +/- 271 and 3,082 +/- 282 cells/m(2), respectively. No statistically significant difference of endothelial cell density between confocal and specular microscopy was found. Endothelial cell count was significantly lower in the automated than in the semi-automated and manual analysis both with confocal and with specular microscopy. In conclusion, endothelial cell count measurements with confocal and contact specular microscopy are comparable. PMID- 12592046 TI - Results of anterior chamber clearance via the pars plana in pseudophakic endophthalmitis. AB - PURPOSE: Patients with endophthalmitis have extensive fibrinous exudates in the anterior chamber. Clearance of these exudates in pseudophakic patients during vitrectomy is difficult. We evaluated the results of the pars plana approach for clearing these exudates. MATERIAL AND METHODS: Eighteen patients with pseudophakic endophthalmitis underwent clearance of anterior chamber exudates through the pars plana approach after initial creation of a peripheral iridectomy with a vitrectome. RESULTS: The exudates were adequately cleared in all 18 patients with good posterior segment visualization during vitrectomy. Minor punctate iris bleeds (4 patients), intraoperative dislocation of the intraocular lens (IOL) (1 patient) and postoperative dislocation of the IOL following blunt trauma, 9 months after vitrectomy (1 patient) were the complications noted. CONCLUSION: This technique is simple, safe and helps clearing anterior chamber exudates. It also permits excellent visualization during vitrectomy for pseudophakic endophthalmitis. PMID- 12592047 TI - Surgical management of epiretinal membrane with indocyanine-green-assisted peeling. AB - PURPOSE: To evaluate the surgical results of pars plana vitrectomy with peeling of the inner limiting membrane (ILM) in a preliminary series of 28 eyes with macular epiretinal membrane (ERM). METHODS: Pars plana vitrectomy with peeling of the ERM was performed on 28 eyes. All preretinal macular tissues were removed including the ILM after injection of indocyanine green (ICG) solution. RESULTS: Postoperatively visual acuity improved within 3 months by at least 2 lines in 26 eyes. Neither the deterioration of the macula nor the recurrence of the ERM was observed during the entire period of review (6 months; range 4-8 months). CONCLUSIONS: Vitrectomy including ICG-assisted peeling of the ERM leads to an expedited resolution of the macular pucker and improvement of visual acuity, without subsequent formation of ERMs. PMID- 12592048 TI - ABCA4 sequence variants in Chinese patients with age-related macular degeneration or Stargardt's disease. AB - ABCA4 gene sequence alterations cause Stargardt's disease (STGD) and may cause some age-related macular degeneration (AMD). We sought to shed light on these associations among Hong Kong Chinese by genotyping 140 AMD, 18 STGD and 95 normal control subjects for 15 ABCA4 exons which were reported to often contain AMD- or STGD-associated mutations. Sequence alterations R212H, T1428M, V1433I, T1572M, I2166M, IVS6-5T>G and IVS33+1G>T were found in AMD patients. T1428M and R2040X occurred in STGD patients. Control subjects displayed all the above missense alterations but no splicing or nonsense changes. Therefore, ABCA4 splicing mutations may be associated with a small proportion of AMD cases. PMID- 12592050 TI - Comparison of antiallergic drugs in an experimental model of ocular anaphylaxis. AB - PURPOSE: To compare the efficacy of antiallergic drugs in an experimental ocular anaphylaxis model. MATERIALS AND METHODS: Fifty-two rats were divided into 8 groups. Dithiothreitol and ovalbumin (OVA) were applied to both eyes of previously OVA-inoculated rats in groups 1-5 and 7. In groups 1-5, antiallergic efficacy of the drugs (lodoxamide, sodium cromoglycate, levacobastine, cimetidine, and diclofenac sodium) instilled on the right eyes were compared with the buffer instilled on the left eyes, measuring the amount of extravasated Evans blue (EB) dye by spectrophotometric analysis. Group 6 served as a control of EB dye extravasations in nonimmunized eyes. Histopathologic examinations for anaphylaxis and controls were done in groups 7 and 8. RESULTS: Only levacobastine and lodoxamide decreased vascular permeability at a significant ratio (46%, p < 0.01 and 31%, p < 0.05, respectively). CONCLUSION: Levacobastine and lodoxamide but none of the three other drugs mentioned above were found to be effective in decreasing vascular permeability. PMID- 12592049 TI - Familial high myopia linkage to chromosome 18p. AB - A locus for autosomal dominant high myopia was reported on chromosome 18p. We sought to confirm this finding and narrow the reported interval by analyzing high myopia among families of Hong Kong Chinese, in whom myopia is common. In 15 families with a possibly autosomal dominant inheritance of high myopia (>or=-6 dpt) in at least 2 generations, 10 chromosome 18p markers were analyzed for linkage with high myopia. Two-point linkage analysis showed trends toward linkage of markers D18S476 and D18S62 with high myopia, with maximum logarithm of odds (LOD) scores of at least 1.1 and 1.7, respectively. Multipoint analysis of those 2 markers gave a maximum LOD score of at least 2.1. To attempt to account for likely genetic heterogeneity, 5 families showing evidence of linkage of the 2 markers with high myopia were selected for further multipoint linkage analysis, resulting in a maximum LOD score of 2.4 at D18S476. While multiple genetic and environmental factors likely contribute to myopia, these data are consistent with the possibility of a locus on chromosome 18p. PMID- 12592051 TI - Interaction of intraocular lenses with various concentrations of silicone oil: an experimental study. AB - The aim of this study was to evaluate the interaction between various widely used intraocular lenses (IOLs) and silicone oils of different viscosities. Four groups of IOLs, including monoblock foldable hydrophilic acrylic IOLs (Morcher, type 92s); monoblock hydrophobic acrylic IOLs (Acrysof-SA60AT, Alcon); single-piece rigid polymethylmethacrylate (PMMA) IOLs (Intraocular Optical International-IOI 65130) and a three-piece foldable silicone optic IOL (CeeOn Edge 911A, Pharmacia UpJohn) were analyzed in vitro to determine the percentage adherence 1,000 centistoke, 1,300-centistoke or 5,000-centistokes silicone oil on the IOL optic. For each IOL type, there was no statistically significant difference in the mean silicone oil coverage (MSC) of the IOL optics for the different viscosities of silicone oil. Silicone IOLs had the highest MSC percentage (79.9%) whereas hydrophilic acrylic IOLs were the least silicone-covered IOLs (7.8%) compared to the other IOL types tested in this study. It is not the concentration of silicone oil that affects silicone oil coverage. When performing small-incision cataract surgery in patients who may require silicone oil injection, foldable hydrophilic acrylic or hydrophobic acrylic lenses should be preferred over standard foldable silicone lenses. PMID- 12592052 TI - Safety of unoprostone isopropyl 0.15% ophthalmic solution in patients with mild to moderate asthma. AB - PURPOSE: To assess the safety of unoprostone isopropyl 0.15% ophthalmic solution (UIOS) in patients with mild to moderate asthma. METHODS: Randomized, double masked, two-period crossover, placebo-controlled study comparing unoprostone 0.15% with vehicle-placebo. Pulmonary function tests conducted at baseline and during the 4-hour postdose observation period included forced expiratory volume in 1 s, forced vital capacity, peak expiratory flow rate, and bronchodilator response after 200 microg inhaled salbutamol. RESULTS: There were no significant changes in pulmonary function up to 4 h postdose after treatment with 1 drop unoprostone 0.15% in each eye. CONCLUSION: Unoprostone 0.15% was safe for use in patients with mild to moderate asthma, without affecting the response to bronchodilator therapy. PMID- 12592053 TI - Color Doppler imaging of choroidal circulation in patients with asymmetric age related macular degeneration. AB - We aimed at evaluating the possible role of choroidal perfusion abnormalities in the development of choroidal neovascularisation (CNV) in patients with age related macular degeneration (AMD). Twenty-six patients who had non-exudative AMD in the first eye and CNV secondary to AMD in the fellow eye were enrolled. Blood flow velocities, vessel pulsatilities and resistivities were measured from ophthalmic artery, nasal and temporal posterior ciliary arteries using colour Doppler imaging. Systolic and diastolic velocities were lower in eyes with CNV for all vessels, except for the systolic velocity of the nasal posterior ciliary artery (p >0.05). Pulsatility and resistivity indices were higher in eyes with CNV for all vessels. This difference was statistically significant for the resistivity index of the nasal and temporal posterior ciliary arteries (p = 0.032 and p = 0.021, respectively) and the pulsatility index of the nasal posterior ciliary artery (p = 0.035). We have shown that in patients with AMD choroidal blood flow is more impaired in the eyes with CNV than in the fellow eyes. PMID- 12592054 TI - Evaluation of treatment by pulsed electromagnetic fields in a rabbit hyphema model. AB - PURPOSE: Previous clinical studies have suggested a positive effect of low frequency pulsed electromagnetic fields (PEMF) on wound healing and inflammation in the eye. We tried to test the value of PEMF treatment in a hyphema animal model with well-defined conditions. METHODS: After injection of citrated autologous blood for the production of hyphema, 16 rabbit eyes were treated with 10 or 20 mT for 60 min on 4 days within the postoperative week. Two control groups with hyphema alone (n = 8) and PEMF irradiation without hyphema (n = 4) were also included. The rate of resorption was recorded daily. Histopathologic evaluation was performed. RESULTS: The incidence of endothelial cell damage and fibrotic clots was markedly reduced in the 10-mT group while the resorption time of 8 days was identical with the control group. In the 20-mT group, the complication rate and the resorption time was increased versus the control group. CONCLUSIONS: The use of PEMF treatment is of some, however, limited value as it did not reduce the resorption time of hyphema but displayed a dose-dependent, beneficial influence on some serious side effects. Future clinical studies with low-dose PEMF irradiation are justified and should determine the optimal dosage and suitable indications of PEMFs as an adjunctive treatment in ocular inflammation or trauma. PMID- 12592056 TI - Topographic mapping of retinal function with the SLO-mfERG under simultaneous control of fixation in Best's disease. AB - PURPOSE: To introduce the scanning laser ophthalmoscope-evoked mfERG (SLO-mfERG) as a new method to measure focal retinal function. METHODS: Sixty-two healthy individuals and 12 patients with Best's disease were examined. mfERGs were recorded using a scanning laser ophthalmoscope as a stimulator and trigger device (He-Neon 632.8 nm) as well as a fundus-monitoring system (Infrared 730 nm). RESULTS: Amplitudes in the central concentric area were found to be significantly lower in patients with Best's disease than in healthy controls, while no significant differences were found for the more peripheral areas. CONCLUSION: SLO mfERG is a reliable new technique for topographic mapping of retinal function under simultaneous control of fixation. PMID- 12592055 TI - The effect of N-acetyl serotonin on ultraviolet-radiation-induced cataracts in rats. AB - AIMS: To investigate the effects of ultraviolet radiation (UVR) on cataract development in rat lenses and whether or not N-acetyl serotonin has an effect on changes in these lenses. MATERIAL AND METHODS: The study was performed using 5 groups of Sprague-Dawley albino rats, with each group consisting of 15 rats. The 5th group being the control group did not receive any applications, whilst the other 4 groups received a daily dose of 0.2 J/cm(2)/day UVR (305 nm wavelength) for 60 days. A dose of 4 mg/kg/0.1 ml N-acetyl serotonin was injected intraperitoneally to group 1 and group 2 every day and on alternate days, respectively. Group 3 received an intraperitoneal injection of 0.1 ml phosphate buffer solution every day, whilst group 4 received no injection. On the 60th day, an intracardiac withdrawal of blood was performed, after the rats had been anesthetized with ether. Following the withdrawal of blood, the rats were killed using a high dose of ether and their eyes were enucleated. The lens fresh weights, plasma malondialdehyde (P-MDA), erythrocyte glutathione peroxidase (E GSHPx), erythrocyte glutathione reductase, blood reduced glutathione (B-RGSH), erythrocyte catalase (E-CAT), lenticular malondialdehyde, lenticular superoxide dismutase (L-SOD), lenticular glutathione peroxidase (L-GSHPx) and lenticular glutathione (L-GSH) levels were all assessed. RESULTS: The lens fresh weights were determined to be lower in group 1 and in the control group in comparison with the other groups (p < 0.01). Whilst the P-MDA level was found to be lower (p < 0.001), the E-GSHPx level was higher (p < 0.01) in the control group than in the other groups. The E-GSHPx level was higher in groups 1 and 2 than in groups 3 and 4 (p < 0.01). The B-RGSH level was higher in the control group than in the other groups (p < 0.001). The E-CAT level was higher in both the control group and group 1 than in groups 2, 3 and 4 (p < 0.01), whilst it was higher in group 2 when compared to groups 3 and 4 (p < 0.01). The L-SOD levels were found to be higher in the control group and group 1 than in groups 2, 3 and 4 (p < 0.001). Whilst the L-GSHPx levels were determined to be higher only in the control group (p < 0.001), the L-GSH levels were higher in the control group and group 1 than in the other groups (p < 0.001). CONCLUSIONS: In recent years, the depletion of the atmospheric ozone layer has resulted in the penetration of more UVR to the earth, which causes various effects on different tissues of organisms. N-acetyl serotonin, a melatonin precursor, may well be effective in the prevention of the negative effects induced by the UVR upon the lens tissue, in which case the capability of melatonin to capture free radicals as well as its antioxidative properties should be taken into consideration. PMID- 12592057 TI - Ocular complications of the Fernand-Widal triad and its therapy. AB - BACKGROUND: The Fernand-Widal triad (FWT) is the association of non-allergic intrinsic asthma, nasal polyposis and sensitivity to Aspirin. The aim of this paper is to describe the possibility of ocular complications, which we found in 3 cases: in 1 case due to the FWT itself and in the other 2 as a result of corticosteroid treatment. METHODS: Three cases of the FWT with ocular complications were studied, and the pertinent literature was reviewed. RESULTS: No previous description of ocular involvement in individuals with FWT was found in the literature. In the first case, episodes of orbital cellulitis due to superinfected nasal polyposis were found as a complication produced by the disease. In the other 2 cases, corticosteroid treatment created complications: ocular hypertension in both cases and bilateral subcapsular cataracts in 1 case. CONCLUSIONS: Although not rare, the diagnosis of the FWT is often missed. This perhaps explains why no report of ocular complications has yet been published in the literature. Because of the serious clinical consequences, physicians involved in the treatment of these patients should be aware of this diagnosis. PMID- 12592058 TI - [Color duplex ultrasound of the temporal artery: replacement for biopsy in temporal arteritis]. PMID- 12592059 TI - Metoclopramide-induced hyperprolactinemia affects mouse endometrial morphology. AB - Information about the effects of pituitary hyperprolactinemia on endometrium, especially in levels coexisting with absence of amenorrhea in women, is scarce. The interference of high prolactin levels on endometrial morphology was thus investigated in young post-pubertal and adult mice rendered hyperprolactinemic by long-term treatment with metoclopramide (MC). No remarkable differences have been noticed upon light microscopy examination of the endometria comparing young to adult cycling MC-treated mice, except on the max/min diameter ratio, which in young animals was lower than in adults (ANOVA, p < 0.01). Both young and adult MC treated mice presented an increased number of endometrial glands than their respective controls (ANOVA, p < 0.01). However, young MC-treated animals showed the highest values of endometrial thickness index compared to other groups (ANOVA, p < 0.01). Our results indicate that MC-induced hyperprolactinemia causes mouse endometrium proliferation, mainly in young animals. PMID- 12592060 TI - Impaired mixed lymphocyte reaction in intrahepatic cholestasis of pregnancy. AB - OBJECTIVE: To clarify the alteration of maternal-fetal mixed lymphocyte reaction (MLR) in intrahepatic cholestasis of pregnancy (ICP). METHODS: Twenty-two patients with intrahepatic cholestasis of pregnancy and 21 normal pregnant women were recruited into the present study. Of patients with ICP, 5 also experienced pregnancy-induced hypertension (PIH). Maternal-fetal mixed lymphocyte reaction was conducted by using a morphologic transformation method, and the transformation rates of lymphocytes were compared either between ICP patients with and without pregnancy-induced hypertension, or between control and ICP group. RESULTS: In the ICP group, the transformation rates of lymphocytes were not significantly different between pregnancies with and without pregnancy induced hypertension (p > 0.5). Compared with control, the transformation rate of lymphocytes in intrahepatic cholestasis of pregnancy decreased significantly (p < 0.001). CONCLUSION: Suppressed mixed lymphocyte reaction was observed in intrahepatic cholestasis of pregnancy and its mechanism behind and role in the disease need to be further explored. PMID- 12592061 TI - Effects of long-term oral hormone replacement therapy on plasma nitric oxide and beta-endorphin levels in postmenopausal women. AB - OBJECTIVES: The aim of this study is to evaluate the relationship between plasma nitric oxide (NO) and beta-endorphin levels in women using hormone replacement therapy (HRT) for 12 months. MATERIAL AND METHODS: Our study group was composed of 55 patients who were in at least their second postmenopausal year. Of the 55 patients, 25 were in the control group. All 30 women in the study group received 2 mg 17beta-estradiol + 1 mg norethisterone acetate tablets daily for 12 months. Plasma NO and beta-endorphin levels were measured both before and after the study period and possible relationships were analyzed. RESULTS: There was a significant increase in both beta-endorphin (p = 0.0001, 10.93 +/- 2.25 vs. 14.85 +/- 2.49) and NO (p = 0.0001, 19.79 +/- 4.01 vs. 27.83 +/- 10.27) levels measured after the study in the HRT group. A correlation was seen between the increments in beta endorphin and NO levels in the HRT group. CONCLUSION: Continuous combined HRT raises both plasma NO and beta-endorphin levels and a close relationship was found between the two molecules after therapy. We postulate that the increment in these molecules may explain some of the beneficial effects of HRT on cognitive function and mood. PMID- 12592062 TI - Body composition modulates the effects of hormone replacement therapy on growth hormone and insulin-like growth factor-I levels in postmenopausal women. AB - OBJECTIVE: To examine the relationships of body composition with basal serum estrone, estradiol, androstenedione, cortisol, growth hormone (GH) and insulin like growth factor-1 (IGF-1) levels in 73 postmenopausal women. DESIGN: Cross sectional study of hormone levels and body composition determined by dual-energy X-ray absorptiometry and anthropometry in women who were not taking oral hormone replacement therapy (HRT) and women taking HRT. Because high adiposity may modify hormone levels, subjects were grouped by fatness into obese (BMI >25 kg/m(2) and waist circumference >80 cm) and lean groups, as well as by HRT use. RESULTS: Total levels of estrone, estradiol, GH and cortisol were significantly higher and IGF-1 was lower in HRT users. In HRT users, estradiol levels were higher and GH levels were lower in obese than lean women. IGF-1 levels were lower in obese HRT users than lean nonusers. Total cortisol levels were significantly higher in lean HRT users than lean nonusers and obese users. GH and IGF-1 were significantly inversely correlated with trunk fat and percent body fat. Multiple regression revealed that only trunk fat was a significant (negative) determinant of GH and IGF-1 levels, whereas HRT use positively and negatively predicted GH and IGF-1, respectively. Percent body fat significantly predicted estradiol levels. Body composition did not differ by HRT use. CONCLUSIONS: Our results suggest that trunk fat attenuates the HRT-induced increase on GH levels. In addition, trunk fat is a significant determinant of low IGF-1 levels in postmenopausal women, and IGF-1 levels decline more with HRT use. PMID- 12592063 TI - Serum estradiol levels after 4 days of ovarian hyperstimulation in oocyte donors are predictive of embryo quality and clinical outcomes. AB - OBJECTIVE: To determine whether estradiol (E(2)) levels after 4 days of ovarian hyperstimulation in leuprolide-downregulated cycles are predictive of embryo quality and pregnancy outcome in oocyte donation. MATERIAL AND METHODS: Retrospective analysis of 287 consecutive oocyte donation cycles performed at a university assisted reproduction therapy program between January 1996 and December 2000. Oocyte donors and recipients followed a non-varied standard synchronization regimen. A serum E(2) (day-5 E(2)) was obtained from egg donors the morning after leuprolide acetate downregulation and 4 days after hMG/FSH administration. Day-5 E(2) was categorized into 3 groups: group I <75 pg/ml; group II 75-350 pg/ml, and group III > or =350 pg/ml. All embryo transfers were performed 3 days after oocyte recovery. RESULTS: Elevated day-5 E(2) levels were directly correlated with higher average embryo scores, greater number of oocytes retrieved, and greater number of embryos available for transfer and cryopreservation. Improved pregnancy outcomes were also noted for group III (day 5 E(2) level >350 pg/ml). CONCLUSION: After 4 days of gonadotropin stimulation, an E(2) level of >350 pg/ml is most predictive of higher embryo grade and pregnancy outcome in oocyte donation. PMID- 12592064 TI - Correlations between extent and spread of adenomyosis and clinical symptoms. AB - OBJECTIVE: To correlate the histopathology of adenomyosis particularly the depth and spread of adenomyosis and symptomatology. STUDY DESIGN: Medical records of 94 patients who had undergone a hysterectomy and who were found to have adenomyosis on histopathologic examination were reviewed and histopathologic slides were reexamined. The symptoms were correlated with the presence of adenomyosis, the depth of penetration, and the spread of adenomyosis foci. RESULTS: Specimens were stratified according to the degree of adenomyosis penetration into 4 groups: group A consisted of specimens with adenomyosis penetration into the myometrium of up to 25%; group B, 26-50%; group C, 51-75%, and group D, >75%. There was a significant correlation between the depth of penetration and the number of adenomyosis foci (r = 0.3446; p = 0.0001). Hemosiderin deposition was found mainly in the specimens with penetration of >75%. The symptoms did not correlate with the degree of penetration (r = 0.088; p NS). However, the spread of adenomyosis correlated significantly with pelvic pain (r = 0.80, p = 0.02), and with dysmenorrhea (r = 0.81, p = 0.01), but not with menorrhagia or dyspareunia. CONCLUSIONS: Hemosiderin deposition and adenomyosis foci are found predominantly in specimens with deep adenomyosis penetration. It suggests that the deeper the penetration, the more extensive the adenomyosis. Symptoms of adenomyosis do not correlate with the depth of penetration, but there is a correlation between the spread of adenomyosis and pelvic pain, and dysmenorrhea. PMID- 12592065 TI - Does the introduction of new technology change population demographics? Minimally invasive technologies and endometrial polyps. AB - OBJECTIVE: To determine if the characteristics of women with endometrial polyps changed as minimally invasive modalities for diagnosis and surgical treatment were integrated to our clinical practice. MATERIALS AND METHODS: The pathologic reports of all women diagnosed with endometrial polyps in 1990 (220 patients) and 1996 (386 patients) were reviewed. The respective medical records were reviewed for anthropomorphic factors, bleeding pattern, diagnosis, and modality of surgical removal. Comparisons of dichotomous data and analysis of nominal variables with two or more categories were carried out with the chi(2) test. RESULTS: Differences in patient characteristics or pattern of bleeding were minimal. In both groups, endometrial polyps were more frequent in women with abnormal bleeding, reaching almost 80%. Diagnostic and therapeutic modalities were significantly different in 1990 vs. 1996. Diagnosis by ultrasound increased fivefold (3.6 vs. 16.8%) and operative hysteroscopy increased threefold (6.4 vs. 19.7). The frequency of incidental diagnosis of endometrial polyps at the time of hysterectomy decreased significantly. The most frequent modality of surgery done in 1990 was dilatation and curettage (44.6%), followed in frequency by endometrial biopsy and abdominal hysterectomy. In contrast, the most frequent modality in 1996 was hysteroscopic resection (36.8%), followed by dilatation and curettage (12.8%). CONCLUSIONS: The introduction of minimally invasive methods of diagnosis and treatment of endometrial polyps has not changed the overall patient population with this lesion. PMID- 12592066 TI - Preoperative findings in non-gynecologic carcinomas metastasizing to the ovaries. AB - This study evaluated whether preoperative findings are useful prognostic indicators in patients with non-gynecologic carcinomas metastasizing to the ovaries. Fifty-three patients were analyzed. Primary sites of carcinoma were discovered after gynecologic surgery in 20 patients. In 10 patients, primary sites of carcinoma were discovered during gynecologic surgery. Thirty-one patients received surgery on the primary site before gynecologic surgery. The primary site could not be identified in 2 patients. The 5-year survival rate was 40.0% in patients, who underwent surgery on the primary site before gynecologic surgery. However, the 5-year survival rate was 12.2% in patients, who underwent surgery on the primary site during or after gynecologic surgery. The CEA level was significantly lower in the stomach primary group than in the intestine primary group. However, CA125 was the only marker significantly affecting survival. In comparison with primary ovarian carcinoma, the sensitivity of CEA was significantly higher for non-gynecologic ovarian carcinoma, while that of CA125 was significantly less sensitive for non-gynecologic ovarian carcinoma. In conclusion, the prognosis was better in patients, who had undergone resection of primary carcinoma before gynecologic surgery on non-gynecologic ovarian carcinoma. CEA is a useful marker to distinguish non-gynecologic from primary ovarian carcinoma, while CA125 is the only significant prognostic marker for non gynecologic ovarian carcinoma. PMID- 12592067 TI - Ureteral and sigmoid obstruction caused by pelvic actinomycosis in an intrauterine contraceptive device user. AB - We report herein a rare case of ureteral and sigmoid obstruction caused by pelvic actinomycosis in a patient fitted with an intrauterine contraceptive device (IUCD). A 63-year-old Japanese woman was admitted complaining of lower abdominal pain and slight fever continuing for a month. She had a history of IUCD insertion 30 years previously and had been menopausal for the past 10 years. Ultrasonography and CT scan revealed a solid pelvic mass involving the uterus, sigmoid colon, urinary bladder, and right ureter. The IUCD was detected in the uterine cavity. Right hydronephrosis and hydroureter due to an obstruction of the distal ureter and the extensive stenosis of the sigmoid colon were also observed. Blood analysis showed leukocytosis, thrombocytosis, and elevated C-reactive protein levels. Although pathological and microbiological analysis of the removed IUCD showed negative results for Actinomyces infection, these findings suggested a pelvic abscess caused by actinomycosis. Benzyl penicillin administration was started immediately. Total hysterectomy, bilateral salpingo-oophorectomy, and lysis of adhesion around the ureter were performed. Actinomycosis was diagnosed based on histologic examination. The patient's postoperative course was uneventful except for persistent mild hydroureter and hydronephrosis. The patient is now healthy without evidence of recurrent Actinomyces infection 1 year after treatment. As shown in the present case, pelvic actinomycosis should be considered as a cause of pelvic inflammatory disease in IUCD users, even though Actinomyces was not detected on the IUCD. PMID- 12592068 TI - Axillary lymphadenopathy as the primary presentation of primary peritoneal carcinoma. AB - The finding of isolated distant metastatic axillary lymphadenopathy without remarkable intra-abdominal disease is very unusual. We report on a patient with primary peritoneal serous carcinoma who initially presented with axillary lymphadenopathy, and was misdiagnosed as having primary breast cancer. In the case of an isolated axillary adenocarcinoma various kinds of immunohistochemical tests should be performed in order to elucidate the origin of the malignancy. PMID- 12592070 TI - Significant risk of a second melanoma in patients with a history of melanoma but no further predisposing factors. AB - It is a well-accepted fact that melanoma patients are prone to develop further melanomas. In a cohort of 535 melanoma patients, we determined the incidence of second melanomas, as well as risk factors known to be associated with melanoma development. A retrospective analysis of patients regularly consulting our melanoma care unit revealed second or even further melanomas in 5.6% (n = 30). When compared to the overall population, melanoma patients have at least a 30 fold increased risk for the development of a second melanoma. Twenty of these 30 patients revealed none of the known predisposing conditions for multiple melanomas. PMID- 12592071 TI - Cutaneous granulomas and malignant lymphomas. PMID- 12592072 TI - Sweet's syndrome and leukemia cutis: a common skin homing mechanism? PMID- 12592073 TI - Steroidogenic isoenzymes in human hair and their potential role in androgenetic alopecia. AB - Androgenetic alopecia (AGA) is the most common type of hair loss. The relatively strong concordance of the degree of baldness in fathers and sons is not consistent with a simple Mendelian trait, and a polygenic basis is considered to be most likely. So far, the predisposing genes for AGA are unknown and we do not understand the molecular steps involved in androgen-dependent beard growth versus androgen-dependent hair loss, but AGA can be defined as a dihydrotestosterone (DHT)-dependent process with continuous miniaturization of sensitive hair follicles. The type 2 5alpha-reductase plays a central role by the intrafollicular conversion of testosterone to DHT. However, due to the increasing knowledge in this field, we now know that there are many more steroidogenic enzymes involved in the onset and development of AGA, and this article shall provide a critical overview of recent discoveries. PMID- 12592074 TI - The role of innate immunity in the pathogenesis of acne. AB - Acne is a multifactorial disease of the pilosebaceous follicle. The most significant pathogenetic factors of acne are: abnormal ductal keratinization, increased sebum secretion, abnormalities of the microbial flora and inflammation. The pilosebaceous unit is an immunocompetent organ. Keratinocytes and sebocytes may act as immune cells capable of pathogen recognition and abnormal lipid presentation, and they might have an important role in initiating and perpetuating the activation of both innate and adaptive immune responses. The elements of the skin immune system are involved in the development of both noninflammatory and inflammatory acne lesions. PMID- 12592075 TI - Angioedema and oral contraception. AB - BACKGROUND: Oral contraceptives can precipitate attacks of hereditary angioedema (ANE) or induce acquired forms. OBJECTIVE: We studied 5 patients who had an ANE which had begun under oral contraception and disappeared after stopping the pill. METHODS: We explored the clinical and biological characteristics of these patients. RESULTS: The symptoms developed during the first year or later after starting contraception; the patients reported relapsing swelling of the lips, hands, larynx and abdomen. All women had normal serum C4 and C1 inhibitor (C1Inh) antigen levels, but a lowered C1Inh activity, with a marked protein cleavage on the immunoblot. The suppression of the pill was associated with the regression of the edema and normalization of C1Inh function. CONCLUSION: The mechanism of these ANE is unknown. The could be due to a modulation of C1Inh expression upon androgens or an imbalance between coagulation proteins favoring C1Inh cleavage by its target proteases. PMID- 12592076 TI - Premenstrual deterioration of skin symptoms in female patients with atopic dermatitis. AB - BACKGROUND: Although it has been recognized that women with atopic dermatitis often show menstrual-cycle-associated skin deterioration, information about this subject is meager. OBJECTIVE: To clarify the clinical features of the monthly worsening of atopic dermatitis. METHODS: A total of 286 Japanese women with atopic dermatitis were interviewed to see whether the menstrual cycle had any influence upon their skin lesions, and whether they had symptoms of premenstrual syndrome. Patients suffering from monthly skin deterioration were then observed during and after the monthly worsening. RESULTS: Of the 286 patients interviewed, 134 (47%) had monthly skin deterioration, most of which occurred in the premenstrual week. There was individual difference in severity of the monthly skin worsening. All patients with the premenstrual skin aggravation had symptoms of premenstrual syndrome. CONCLUSIONS: Premenstrual worsening of skin lesions occurs in approximately half of women with atopic dermatitis. The premenstrual skin deterioration is related to the premenstrual syndrome. PMID- 12592077 TI - Chronic pemphigoid gestationis: comparative clinical and immunopathological study of 10 patients. AB - BACKGROUND: Pemphigoid gestationis (PG) is a rare autoimmune bullous disorder occurring during the last trimester of pregnancy and usually regressive within 3 months after delivery. Prolonged forms of the disease lasting more than 6 months after delivery have been reported as chronic PG. OBJECTIVE: The aim of the present study was to compare the clinical and immunopathological findings between 4 patients presenting a normal regression of the disease after delivery and 6 patients with a chronic course. METHODS: All patients were evaluated and studied by clinical patterns (age, mucosal and cutaneous involvement, obstetrical history, duration of the blistering disease and response to treatment), by direct and indirect immunofluorescence and Western blot. Eight patients were studied by immunoelectron microscopy (IEM) and 3 patients had an indirect IEM. RESULTS: Patients with chronic PG were older, had multigravidity, a history of PG during previous pregnancies, widespread cutaneous eruption and mucosal involvement. Subclass analysis of circulating autoantibodies showed an IgG1 anti-BP180 response in all patients except 1 with disease of 7 years' duration. Direct IEM was positive in 6/8 patients showing a labeling of the lamina lucida, and indirect IEM using colloidal gold probes confirmed the localization of the target antigens to the proximal part of the anchoring filaments in the lamina lucida. CONCLUSION: This study suggests that, even in chronic long-lasting PG, IgG1 remains the predominant subtype of IgG. Therefore no biological and predictable marker of chronicity can be ascertained from this series. PMID- 12592078 TI - Lupus miliaris disseminatus faciei: a distinctive rosacea-like syndrome and not a granulomatous form of rosacea. AB - BACKGROUND: Lupus miliaris disseminatus faciei is an eruption of discrete red brown, dome-shaped papules, histologically characterized by epithelioid cell granulomas. The pathogenesis of the disorder remains controversial. OBJECTIVE: The authors discuss the place of lupus miliaris disseminatus faciei among granulomatous disorders. METHODS: This report reviews the available literature and presents 3 patients with lupus miliaris disseminatus faciei. RESULTS: The histopathologic characteristics of lupus miliaris disseminatus faciei--an epithelioid cell granuloma with central necrosis--may be found in granulomatous rosacea, whilst the clinical features and course of lupus miliaris disseminatus faciei are often similar to cutaneous sarcoidosis. CONCLUSIONS: We put forward lupus miliaris disseminatus faciei as a distinctive rosacea-like syndrome and not as a granulomatous form of rosacea. PMID- 12592079 TI - Mal de Meleda: a new geographical localization in Anatolia. AB - BACKGROUND: Mal de Meleda (MDM) is an autosomal recessive form of palmoplantar keratoderma first described on the Dalmatian island of Meleda. MDM has been observed in many other countries so that the origin of the MDM gene may be elsewhere than in Meleda. OBJECTIVE: After identification of the first MDM patient during a medical visit, a study was planned to reveal other families with MDM in the Koprucay region in Anatolia. METHODS: The patient was interviewed with a questionnaire including a pedigree drawing. All the subsequent cases reported to be of MDM were visited for clinical examination and pedigree drawings. RESULTS: Thirty-nine patients, 8 families and 2 additional cases with MDM were identified in a 50-km(2) mountainous region in Koprucay canyon in Anatolia. The prevalent clinical features were nail involvement (80%), 'glove-and-sock' distribution of the keratoderma (60%), edema on the hands/feet (60%), conical tapering of the fingertips (60%) and hyperhidrosis on the palms and soles (50%). CONCLUSION: To our knowledge, this is the largest series reported. The relationship between the patients in Meleda and those in Anatolia awaits discovery by further researches that will be carried out with the collaboration of dermatology, genetics and medical history departments. PMID- 12592080 TI - Lipid composition of outer stratum corneum in hereditary palmoplantar keratodermas. AB - BACKGROUND: The analysis of lipid composition of the outer stratum corneum is a promising approach to study the pathophysiology of inherited disorders of keratinization. OBJECTIVE: The purpose of the study was the search for biochemical alterations of stratum corneum lipids in hereditary palmoplantar keratoderma (PPK). METHODS: Using high-performance thin-layer chromatography, we performed an analysis of all major stratum corneum lipid classes in scales of 29 patients with 8 different types of hereditary PPK. RESULTS: In comparison to the controls, slight differences in the lipid pattern were found in all keratodermas. Reduced amounts of total ceramides and increased levels of free fatty acids were noted in nearly all types. CONCLUSIONS: The study indicates that the abnormal composition of stratum corneum lipids in PPK is probably not caused by genetic defects of the epidermal lipid metabolism, but it appears to represent an epiphenomenon of a disturbed cornification. PMID- 12592081 TI - Microbiological aspects of diaper dermatitis. AB - BACKGROUND: The microbiological basis of diaper dermatitis is not clearly elucidated, although a better knowledge of microbial colonisation can be of importance with regard to an adequate treatment. OBJECTIVE: To investigate the relevance of candida sp. and Staphylococcus aureus colonisation in diaper dermatitis and to determine the correlation between the extent of colonisation and the severity of disease. METHODS: Growth of candida sp. and S. aureus in the perianal, inguinal and oral regions was determined by positive/negative and semi quantitative analysis in an open, multi-centre (n = 3) study. Forty-eight children with healthy skin and 28 with diaper dermatitis were analysed. The severity of diaper dermatitis was assessed using a total symptoms score. RESULTS: Colonisation by candida sp. was significantly more frequent in children with diaper dermatitis as compared to those with healthy skin (perianal 75 vs. 19%; inguinal 50 vs. 10%; oral 68 vs. 25%, p < 0.0003), whereas colonisation by S. aureus at the 3 swab locations was not different (p > 0.34). There was a highly significant, positive correlation between severity of disease and extent of candida sp. colonisation at all swab locations. CONCLUSIONS: Limited microbial colonisation in diaper dermatitis is of questionable relevance, but extensive colonisation seems to aggravate the symptoms; therefore, we suggest that semi quantitative evaluation should be preferred to the positive/negative assessment for a differential diagnosis. PMID- 12592082 TI - Bexarotene is a new treatment option for lymphomatoid papulosis. AB - BACKGROUND: Lymphomatoid papulosis (LyP) is a clonal T cell proliferation with large cell histology, a chronic course, and an increased risk of lymphoma. Bexarotene (Targretin) is an RXR-selective retinoid (rexinoid) approved for the cutaneous manifestations of cutaneous T cell lymphoma. OBJECTIVE: To determine whether bexarotene is effective in treating LyP. METHODS: Ten patients with chronic and symptomatic LyP were prospectively treated with oral (n = 3) or topical gel (n = 7) formulations of bexarotene. RESULTS: A favorable response to bexarotene treatment with decreased numbers or duration of lesions was seen in all with objective responses in 8 patients. CONCLUSIONS: Bexarotene may be an effective palliative treatment for LyP, warranting further controlled studies. PMID- 12592083 TI - CO2 laser treatment of warts in immunosuppressed patients. AB - BACKGROUND: A large proportion of chronically immunosuppressed patients suffer from recalcitrant viral warts. Furthermore, the incidence of nonmelanoma skin cancer is clearly elevated in this patient group and human papillomavirus is most likely to play a causal role in these neoplasms. OBJECTIVE: To analyze the effectiveness of treatment of viral warts with the CO(2) laser in immunosuppressed patients. METHODS: Multiple viral warts of 13 patients with immunosuppression, a majority of them renal allograft recipients, were treated with CO(2) laser in local or regional anesthesia. In case of recurrences, the lesions were treated again after 3 months. RESULTS: 12 of 13 patients experienced dramatic improvement after 1-3 treatment sessions, 6 of these patients showed full remission. Side effects such as postoperative pain or scarring were minimal. CONCLUSION: CO(2) laser treatment of recalcitrant warts in immunosuppressed patients shows promising results which are comparable to those in immunocompetent hosts. This offers a welcome enlargement of the array of therapies in this otherwise difficult-to-treat group of patients and may contribute to the reduction of risk factors for nonmelanoma skin cancer. PMID- 12592084 TI - Treatment of psoriatic nails with topical cyclosporin: a prospective, randomized placebo-controlled study. AB - BACKGROUND: Nail involvement is a frequent event in the course of psoriasis causing severe distress. While systemic cyclosporin (CsA) represents a well established therapy of psoriasis, its topical use is limited by the difficult penetration of the molecule through the skin and the nail because of its highly lipophilic nature. OBJECTIVES: We carried out a prospective randomized placebo controlled study in order to analyze the effectiveness and tolerability of topical oil-dissolved 70% CsA solution in nail psoriasis. METHODS: Sixteen adult patients with nail psoriasis, divided randomly into two groups of 8 patients (group A and group B), were treated respectively with a 70% maize-oil-dissolved oral CsA solution and maize oil alone. To compare the therapeutic effectiveness, all patients were evaluated, before starting the treatment and after 12 weeks of therapy, by the same dermatologists. The patients were also asked to assess the severity of their nail involvement at baseline and at the end of the treatment. RESULTS: In group A, 3 patients came to a complete resolution of nail lesions and 5 showed a substantial improvement of the overall severity score. In group B, a slight improvement was noted in only 1 patient. All the patients of group A judged positively the results of the therapy, while in group B only 1 patient reported a moderate improvement. CONCLUSION: Our results show that topical therapy with oral CsA solution is a safe, effective and cosmetically highly acceptable treatment modality for nail psoriasis. The ability of CsA to influence keratinocyte proliferation and T-cell lymphokine release, reducing the cornification of the upper layers of the epidermis, may prevent the typical alterations observed in nail psoriasis. PMID- 12592085 TI - Cutaneous granulomas associated with adult T-cell leukemia/lymphoma. AB - A 59-year-old Japanese woman showed recurrent violaceous, indurated erythemas and papules on the buttocks and extremities for a period of 7 years. The lesions showed histologically epithelioid cell granulomas associated with lymphocytes. After 7 years, the patient developed adult T-cell leukemia (ATL), of the acute type, the course of which was fatal. Immunohistochemical staining of the skin sections of granulomatous lesions showed prominent infiltration of CD25-positive cells. Human T-lymphotropic virus type I proviral DNA and monoclonal T-cell receptor beta gene rearrangement were detected in the skin samples by a retrospectively performed gene analysis. We believe that the patient initially had an unusual cutaneous type of ATL in which granulomas occurred as a host protective response against ATL progression. PMID- 12592086 TI - Microvenular hemangioma. AB - Microvenular hemangioma is a rare, slowly growing, benign vascular tumor. It usually presents as a solitary, asymptomatic, purple to red papule or plaque in young to middle-aged adults. Histologically, the tumor is composed of irregular, branching venules with inconspicuous lumina with a lack of endothelial atypia. We describe a case of microvenular hemangioma in a 40-year-old female. The lesion was slightly tender unlike that of previously reported cases. In the context of the histological similarity to a low-grade malignant tumor, Kaposi's sarcoma, the awareness of microvenular hemangioma, a benign vascular tumor, is important. PMID- 12592087 TI - Papular xanthoma associated with angiokeratoma of Fordyce: considerations on the nosography of this rare non-Langerhans cell histiocytoxanthomatosis. AB - BACKGROUND: Papular xanthoma (PX) is a rare normolipidemic non-Langerhans cell histiocytoxanthomatosis affecting both children and adults. OBJECTIVE: We describe an adult case of PX associated with angiokeratoma of Fordyce and review the literature in order to compare and discuss previous reports. METHODS: We studied the clinical, histopathological, immunocytochemical and ultrastructural findings. RESULTS: We report the findings of our case and compare our case with those described in the literature. CONCLUSIONS: Three clinical patterns of PX appeared to emerge in the review of the literature: a self-healing form, a persistent form and a progressive form. The progressive form of PX can be considered the same clinical entity that is also described as progressive nodular histiocytosis. PMID- 12592088 TI - 31-year-old injection drug user with massive skin necrosis and shock. AB - Massive tissue necrosis and septic shock have recently been reported in a series of injection drug users in Scotland, England and Ireland. We report the first case outside the UK meeting the criteria for this new entity (septic shock without fever in an injection drug user, local and systemic inflammation, rhabdomyolysis and tissue necrosis at the injection site). Following surgical treatment and antibiotic treatment, the patient was cured and is currently well. PMID- 12592089 TI - A case of unilateral, systematized linear hair follicle nevi associated with epidermal nevus-like lesions. AB - Multiple hair follicle nevi are an extremely rare condition. In 1998, a case of unilateral multiple hair follicle nevi, ipsilateral alopecia and ipsilateral leptomeningeal angiomatosis of the brain was first reported from Japan. Very recently, hair follicle nevus in a distribution following Blaschko's lines has also been reported. In this paper, we observed a congenital case of unilateral, systematized linear hair follicle nevi associated with congenital, ipsilateral, multiple plaque lesions resembling epidermal nevi but lacking leptomeningeal angiomatosis of the brain. These cases implicate the possibility of a novel neurocutaneous syndrome. Additional cases should be sought in order to determine whether this condition is pathophysiologically distinct. PMID- 12592090 TI - Cutaneous recall reactions with systemic fluorouracil. AB - 5-Fluorouracil is used extensively to treat actinic keratoses as there is selective excessive uptake of 5-fluorouracil in rapidly dividing cells, thus causing inflammation in lesional skin. We report 2 cases of inflammation occurring on clinically normal skin after fluorouracil had been administered parenterally for carcinoma of the oesophagogastric junction and lower third of the oesophagus, respectively. In the first case, actinic keratoses had previously been treated with topical fluorouracil and in the second case, although there were no previous actinic keratoses, there was a high degree of previous solar exposure of the affected areas. In each case, there was a good response to topical steroid application. No recurrence was experienced on further doses of systemic fluorouracil when the skin was treated prophylactically with topical steroids. Recognition of these reactions is important if unnecessary discontinuation of chemotherapeutic drugs is to be avoided as a result of fear of allergic drug reactions. PMID- 12592092 TI - Potential use of leukotriene inhibitors in treatment of psoriasis. PMID- 12592091 TI - Granular parakeratosis in a child. PMID- 12592093 TI - Infliximab: a promising new treatment option for ulcerated necrobiosis lipoidica. PMID- 12592094 TI - The eyelids and metastatic breast carcinoma. PMID- 12592095 TI - Calcitonin receptor-like receptor is expressed on gastrointestinal immune cells. AB - BACKGROUND/AIMS: Pharmacological and morphological studies suggest that the gut mucosal immune system and local neuropeptide-containing neurones interact. We aimed to determine whether gut immune cells are targets for calcitonin gene related peptide (CGRP), which has potent immune regulatory properties. METHODS: Using density gradient centrifugation, rat lamina propria mononuclear cells (LP MNCs) and intra-epithelial lymphocytes (IELs) were isolated. RT-PCR was employed for the detection of mRNA of rat calcitonin receptor-like receptor (CRLR), which is considered to represent the pharmacologically defined CGRP receptor-1 subtype, as well as mRNA of the receptor activity-modifying proteins, which are essential for CRLR function and determine ligand specificity. A radioreceptor assay was employed for the detection of specific CGRP binding sites. RESULTS: RT-PCR and DNA sequencing showed that LP-MNCs and IELs express CRLR. Incubation of isolated LP-MNCs with radiolabelled alphaCGRP revealed the existence of specific binding sites for CGRP. CONCLUSION: These novel data indicate that mucosal immune cells of the rat gut are a target for CGRP and provide significant evidence that CGRP functions as an immune regulator in the gut mucosa. PMID- 12592096 TI - Gastric electrical stimulation in intractable symptomatic gastroparesis. AB - BACKGROUND: The treatment of gastroparesis remains unsatisfactory despite prokinetic and anti-emetic drugs. Gastric electrical stimulation has been proposed as a therapeutic option. We have assessed the effect of gastric electrical stimulation on symptoms, medical treatment, body weight and gastric emptying in patients with intractable symptomatic gastroparesis in a non-placebo controlled study. METHODS: In this multicenter study, 38 highly symptomatic patients with drug-refractory gastroparesis were enrolled. Patients first received temporary electrical stimulation using percutaneous electrodes. The 33 responders to temporary stimulation then underwent surgical implantation of a permanent stimulator. Severity of vomiting and nausea was assessed before and after stimulation. Patients were reassessed 3, 6, and 12 months after permanent implantation. RESULTS: With stimulation, 35/38 patients (97%) experienced >80% reduction in vomiting and nausea. This effect persisted throughout the observation period (2.9-15.6 months, 341 patient-months). Gastric emptying did not initially change, but improved in most patients at 12 months. At 1 year, the average weight gain was 5.5% and 9/14 patients initially receiving enteral or parenteral nutrition were able to discontinue it. CONCLUSION: Electrical stimulation of the stomach has an immediate and potent anti-emetic effect. It offers a safe and effective alternative for patients with intractable symptomatic gastroparesis. PMID- 12592097 TI - Evaluation of the barostat for recordings of gastrointestinal motility. AB - BACKGROUND: The barostat has been used to investigate gastrointestinal motility. No detailed technical evaluation to characterize the properties of this device for motility recordings has been reported. We, therefore, aimed to test the barostat in vitro under standardized conditions. METHODS: Barostat and manometry recordings were performed using a combined catheter in a pressure chamber. Some of the experiments were made in pig sigmoid colon in order to mimic recordings in a hollow organ. Data are mean +/- SD. RESULTS: Baseline changes of the bag volume under constant conditions were 3.9 +/- 2.0%. The bag volume increased by 4.5 +/- 1.1% with a temperature increase from 22 to 37 degrees C (p < 0.05). At external pressures above the bag operating pressure, the barostat bag collapsed, while only minimal volume reductions occurred at external pressures below the bag pressure. Barostat recordings of pressure events were delayed when compared with manometry and not linearly related to the pressure increases. CONCLUSION: The influence of temperature changes on barostat recordings is small. The volume decrease in the barostat bag is not linearly related to the external pressure increase and influenced by the operating pressure in the bag. This experimental study provides insight and caveats for those planning to use the barostat device for motility recordings of the gastrointestinal tract. PMID- 12592098 TI - Cyclooxygenase-2 expression is associated with well-differentiated and intestinal type pathways in gastric carcinogenesis. AB - BACKGROUND/AIMS: Cyclooxygenase-2 (COX-2) expression appears to be increased in several different types of human cancers, suggesting that the presence of COX-2 is associated with carcinogenesis. Recently, increased expression of COX-2 has been frequently detected in gastric cancer, and this may have prognostic significance. In the present study, we aimed to analyze the expression of COX-2 in a much larger sample to determine whether COX-2 expression is related to the clinicopathological features and survival rates of patients with gastric cancer. METHODS: We investigated 140 patients with gastric cancer who underwent surgery between January 1992 and December 1993 and examined the expression of COX-2 in human gastric cancer tissue by immunohistochemistry. RESULTS: COX-2 expression was present in the cytoplasm of tumor cells but not in normal gastric epithelia. Positive expression of COX-2 was detected in 86 of 140 gastric cancers analyzed (61.4%). Positive expression of COX-2 correlated with the depth of tumor invasion (p = 0.015). However, there was no association between COX-2 expression and tumor stage or status of lymph node or distant metastasis. Furthermore, COX-2 expression was not associated with patient survival (p = 0.816). Positive expression of COX-2 occurred more frequently in intestinal than in diffuse or mixed types of cancer and correlated with tumor differentiation (p < 0.001, p = 0.001, respectively). CONCLUSION: These results suggest that COX-2 may play an important role in the evolution of gastric carcinogenesis and be associated with well-differentiated and intestinal type pathways in gastric carcinogenesis. However, COX-2 expression seems to be less useful for establishing prognosis for gastric cancer. PMID- 12592099 TI - Prospective comparison of endoscopy, endosonography and computed tomography for staging of tumours of the oesophagus and gastric cardia. AB - BACKGROUND: Local and multimodal therapeutic strategies for tumours of the oesophagus and gastric cardia, require precise preoperative staging. Endosonography is considered the most accurate staging method, while computed tomography (CT) has limitations especially in the evaluation of local infiltration. Macroscopic endoscopic evaluation was reported to be accurate in selected series, but no study has yet compared all three staging modalities. METHODS: One hundred and seventeen unselected patients with tumours of the oesophagus and gastric cardia were prospectively staged first by the endoscopic macroscopic appearance and then by endosonography. All patients had preoperative CT scans, however, only the 36 patients receiving the scans at our institution were included in the study. The preoperative staging results were then compared to postoperative histology which was available as the gold standard in all included patients. Kappa statistics were used to exclude chance agreement of the clinical staging results with the pathohistological findings. Differences between the resulting kappa values for the different staging modalities were analysed with a jack-knife test. RESULTS: Endoscopic macroscopic staging and endosonography (accuracy 67 and 69%, weighted kappa 0.78 and 0.84) were significantly more accurate than CT (accuracy 33%, weighted kappa 0.44) for determination of the T category (p = 0.006 and p = 0.001). After exclusion of tumours of the cardia (n = 33), the accuracy of macroscopic and endosonographic staging (accuracy 72 and 75%, weighted kappa 0.86 and 0.88) increased and remained more accurate than CT (accuracy 50%, weighted kappa 0.62). The main pitfall in our series in staging the T category was the overestimation of T2 tumours in the cardia as T3 or even as T4 tumours due to the inability to visualise the serosa. The accuracy of predicting lymph node metastasis was 68% for macroscopic endoscopic, 79% for endosonographic, and 67% for CT staging. Only endosonographic staging was significantly different from chance agreement with histology (weighted kappa = 0.56). Endosonographic staging was significantly more accurate than endoscopic macroscopic and CT staging (p = 0.03). CONCLUSIONS: Endosonography is the most accurate staging modality for overall preoperative staging of oesophageal and cardial tumours. Endoscopic macroscopic staging allows a reasonably accurate assessment of the T category. PMID- 12592100 TI - Induction of IkappaB-kinase by cholecystokinin is mediated by trypsinogen activation in rat pancreatic lobules. AB - BACKGROUND AND AIMS: Supramaximal concentrations of cholecystokinin (CCK) or cerulein induce the intracellular activation of trypsinogen and the transcription factor NF-kappaB, a key regulator of inflammatory gene expression. Both events occur early in the development of an acute pancreatitis. The aim of this study was to examine the relationship between intracellular trypsinogen and NF-kappaB activation. METHODS: We detected NF-kappaB-binding activity in electromobility shift assays, IkappaB proteolysis in Western analysis and endogenous IkappaB kinase (IKKalpha and beta) activation using immune complex kinase assays following treatment with CCK in rat pancreatic lobules. To block intrapancreatic trypsinogen activation, a potent and cell-permeable serine-protease inhibitor, Pefabloc, was used. RESULTS: CCK-induced IkappaBalpha degradation and subsequent NF-kappaB activation correlated closely with the catalytic activity of IKKs to phosphorylate IkappaBalpha in vitro. Activation is dose-dependent and peaked at 30 min. Doses of Pefabloc sufficient to inhibit trypsin activation reduced CCK induced activation of NF-kappaB whereas TNF-alpha-induced NF-kappaB activation was not blocked but slightly increased. Moreover, treatment with Pefabloc as well as another serine protease inhibitor, FUT175, inhibited CCK-induced IKK activation. CONCLUSION: These results suggest that intrapancreatic activation of trypsinogen may contribute to NF-kappaB signaling via IKK activation in cerulein pancreatitis. This also explains the fact that only doses of CCK which activate trypsinogen induce NF-kappaB activation in pancreatic acinar cells. Thus, trypsinogen activation is likely to modulate signaling events in acinar cells in the initial phase of acute pancreatitis. PMID- 12592101 TI - Inhibition of intestinal bacterial translocation with rifaximin modulates lamina propria monocytic cells reactivity and protects against inflammation in a rodent model of colitis. AB - BACKGROUND: A modification of the intestinal flora and an increased bacterial translocation is a common finding in patients with inflammatory bowel disease as well as in animal model of colitis. Rifaximin, a non-absorbable derivative of rifamycin, is an effective antibiotic that acts by inhibiting bacterial ribonucleic acid synthesis. AIMS: In the present study, we investigated the effect of the administration of rifaximin (10, 30 and 50 mg/kg/day) or prednisolone (10 mg/kg/day) in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. METHODS: Colitis was induced in mice by intrarectal administration of TNBS (1.5 mg/mouse in 50% ethanol) and disease severity assessed clinically and by histologic scoring of colon damage, determination of interleukin (IL)-2, IL-12, interferon (IFN)-gamma and tumor necrosis factor (TNF) alpha (protein and mRNA and myeloperoxidase (MPO) activity in the colon. Cytokines production by the lamina propria mononuclear cells (LPMC) and luminal bacteria were also measured. RESULTS: Rifaximin administration (30 or 50 mg/kg/day) increased survival rates of colitic mice and reduced colitis severity as demonstrated by improvement of wasting syndrome, histologic scores, decrease in colon IL-2, IL-12, IFN-gamma and TNF-alpha (protein and mRNA) levels, and diminished colon MPO activity. Rifaximin administration caused a significant reduction of colon bacterial translocation towards mesenteric lymph nodes. LPMC obtained from rifaximin-treated mice released significantly lower amount of IFN gamma in response to ex vivo stimulation with agonistic anti-CD3 and anti-CD28 antibodies. Rifaximin (50 mg/kg/day) significantly accelerates recovery in mice with established colitis. CONCLUSIONS: Luminal bacterial microflora plays a role in the pathogenesis of TNBS-induced colitis in mice. Rifaximin administration reduces the development of colitis and accelerates healing of established disease by preventing bacterial translocation. PMID- 12592102 TI - The effect of oxygen supplementation on the arterial ketone body ratio in human cirrhosis. AB - BACKGROUND/AIMS: Data from studies in experimental models have suggested that the impairment of mitochondrial function and altered redox state that occur in cirrhosis may be due to impaired hepatic oxygenation. Since interventions that improve oxygen delivery to hepatocytes may improve mitochondrial functions, we studied the effects of oxygen supplementation on the arterial ketone body ratio (AKBR) in normal volunteers and patients with cirrhosis. METHODS: After a 2-hour fast, ketone bodies were measured in arterial blood taken from patients and controls while breathing room air and then after breathing oxygen via a face mask at 12 liters/min for 60 min. RESULTS: The AKBR was reduced in cirrhotic patients compared with controls, 0.74 +/- 0.23 and 1.51 +/- 0.4, respectively (p = 0.002). Oxygen supplementation significantly improved the AKBR in cirrhotic patients, from 0.74 +/- 0.23 to 1.04 +/- 0.28 (p = 0.001) but did not affect the AKBR in controls. CONCLUSION: These findings suggest that reduced hepatocyte oxygenation contributes to impaired hepatic mitochondrial function in cirrhosis. Strategies at increasing hepatic oxygen delivery may improve hepatic mitochondrial function in patients with chronic liver disease. PMID- 12592103 TI - Mesenteric inflammatory veno-occlusive disease: a rare cause of intestinal ischemia. The first description of recurrent disease. AB - BACKGROUND: Mesenteric inflammatory veno-occlusive disease (MIVOD) is a rare cause of intestinal ischemia of unknown etiology. Histologically, MIVOD is characterized by extended thrombophlebitis and fibrous organized thrombosis of multiple veins. The arteries are by definition not involved. Management includes surgery in all cases described. Recurrence has not been described until now. METHODS: We describe the case of a 64-year-old woman operated with the suspicion of intestinal ischemia. Clinical and histopathological characteristics are reported. RESULTS: The patient underwent a right hemicolectomy and segmental resection of the terminal ileum 15 months later for a recurrence. The histological examination of the resected specimen confirmed the diagnosis of MIVOD and a recurrence thereof. CONCLUSION: MIVOD is a rare cause of colonic ischemia. Recurrence of the disease, which is described here for the first time, is unusual after surgical resection. PMID- 12592104 TI - Training in pediatric critical care medicine: A survey of pediatric residency training programs. AB - BACKGROUND: After completing their critical care rotations, pediatric residents are expected to have acquired skills in the resuscitation of critically ill newborns and children. Recent Accreditation Council on Graduate Medical Education (ACGME) guidelines have limited the time devoted to critical care training during pediatric residency. We sought to determine how individual programs have structured their critical care training experience in light of these changes. MATERIALS AND METHODS: A questionnaire was mailed to each pediatric residency program listed in the 1996-1997 Graduate Medical Education Directory. Information was obtained regarding the structure of critical care training. Data were analyzed using descriptive techniques, one-way analysis of variance with Scheffe post hoc test, and Fisher exact test as appropriate. RESULTS: Data were received from 149 programs (71% response rate). Most programs were in compliance with ACGME standards regarding the number of months devoted to neonatal intensive care, pediatric intensive care, and emergency medicine. There were no significant differences in the total number of rotations in either the neonatal intensive care unit (NICU) or the pediatric intensive care unit (PICU) when the programs were stratified by size. There were no significant differences in the percentage of programs requiring night call in either the NICU or the PICU during off service months. However, small programs (< 25 residents) required significantly fewer rotations in emergency medicine (P < 0.001). Most programs complemented the critical care experience by offering additional rotations and advanced life support training. CONCLUSIONS: Pediatric residency programs have structured their critical care rotations in a similar fashion in accordance with ACGME guidelines. The success in meeting the stated objectives, as measured by the ability of graduating residents to stabilize critically ill children, is not known and will require further study. PMID- 12592105 TI - Midfoot injury in children related to mini scooters. AB - The remarkable rise in popularity of mini scooters has been accompanied by an increase in the number of foot and ankle injuries. We have observed unique pediatric foot injuries related to the use of mini scooters. An association may be present linking the design of the mini scooter and the riding practices of children. We report on two unique midfoot injuries in children occurring with mini scooter use and discuss how these foot injuries may be related to scooter design and children's riding practices. Safety guidelines aimed at the prevention of foot and ankle injuries while riding mini scooters are outlined. PMID- 12592106 TI - Nonurgent pediatric emergency department visits: Care-seeking behavior and parental knowledge of insurance. AB - OBJECTIVES The goals of this study were to describe the factors associated with utilization of emergency services for nonurgent illnesses by insured children in a pediatric emergency department (PED) and to assess parental knowledge of their insurance and its influence on care-seeking behaviors. METHODS We conducted a prospective, descriptive survey of parents of insured children evaluated for nonurgent illnesses in an urban PED. RESULTS A total of 251 surveys were completed. The primary reason chosen by the parent for the PED visit was convenience in 62.8% of cases, a perceived true emergency in 33.6%, and lack of other access to a physician in 3.6%. Parents choosing the PED for perceived emergencies were more likely to state that the illness was life threatening or required hospital admission than those who came because of convenience (33/84 vs. 28/157; < 0.001). Only 38.7% stated that they were educated as to what problems are considered emergencies by their insurance carriers. PED referrals for nonurgent complaints are required by the insurance carrier for 74.9% of the population; however, only 37.2% of the parents stated that referrals were necessary. Primary care physician (PCP) contact was made prior to the visit by 45.4% of parents. Of those who called the PCP, 72.6% stated that they were referred to the PED. Of the parents who believed that a referral was required, those stating that the problem was an emergency were more likely to have contacted the PCP than those who came because of convenience (27/37 vs. 22/46; < P 0.037).CONCLUSIONS Parents frequently do not understand their insurance coverage as it relates to emergency care utilization. This lack of knowledge influences their care-seeking behaviors for nonurgent illnesses. Convenience is a significant factor in PED utilization for nonurgent complaints. PMID- 12592107 TI - Psychosocial needs of young persons who are victims of interpersonal violence. AB - OBJECTIVE: Although most emergency departments (EDs) in the United States treat a large number of patients who are victims of interpersonal violence, few address their psychosocial needs. We conducted this study to determine the needs of victims of interpersonal violence using a psychosocial assessment tool administered in the ED. METHODS: We conducted a descriptive study of a prospective, consecutive random sample of young victims of violence in an inner city, level I adult and pediatric ED with 44,000 annual visits. Youths aged 10 to 24 years who were victims of life- or limb-threatening interpersonal violence in 1998 and 1999, excluding domestic violence, sexual assault, and child abuse, were interviewed using an assessment tool designed to determine the psychosocial needs of young persons. The youth assessment tool was composed of 13 constructs, including exposure to violence, legal issues, drug use and abuse, mental health problems, gang issues, gun accessibility, educational needs, employment, life skills, pregnancy and parenting issues, medical care, and recreational activities. The study was approved by our Institutional Review Board. RESULTS: We interviewed 112 victims of interpersonal violence with a mean age of 18.6 years; 83.9% were male, 67.9% were African American, and 30.4% were Hispanic. Of the youths interviewed, 72.1% had been shot, 15.3% had been stabbed, and 12.9% had been assaulted. In addition, 66.3% had been involved with the law, 73.7% used alcohol, and 65.3% used illegal drugs. Of the interviewees, 41.2% answered yes to at least one question in the CAGE (cut down on your drinking? annoyed by criticism of your drinking? Guilty feeling about your drinking? ye-opener drinking?) assessment, 42.0% reported feeling sad or "down" prior to their injury, and half (54.1%) were involved in a gang. A total of 63.4% of the victims were not in school, did not finish school, or had not obtained a graduate equivalency diploma. The top psychosocial needs were determined to be educational issues (21.8%), occupational issues (ie, job; 16.8%), social issues (eg, gang involvement; 14.9%), and mental health problems (13.2%). CONCLUSIONS: Young persons who are victims of interpersonal violence have a significant need for educational support, employment assistance, mental health services, and gang intervention services. PMID- 12592108 TI - Pediatric emergency medicine fellowship training in the new millennium. AB - OBJECTIVE Fellowship training in pediatric emergency medicine has been available since the early 1980s. Its availability increased rapidly in the late 1980s and early 1990s, but its growth has been much slower in recent years. In this report, we characterize and compare the training programs of today to those that existed 10 years ago. Our study deals with program content and design, focusing on five aspects of fellowship training: demographics, curriculum, clinical emergency department time, research, and benefits. The data gathered in this study are meant to assist programs, both new and old, in enhancing their fellowship training.METHODS A 43-question survey was mailed to all known pediatric emergency medicine fellowship program directors in March of 2000. Two additional attempts were made to obtain survey responses. Forty of the 50 program directors responded, for a response rate of 80%. Statistical analysis was performed, and the data were compared with data that were gathered in two previous studies of fellowship training programs conducted in 1988 and 1991.RESULTS Fellowship training in pediatric emergency medicine continues to grow but at a slower pace than previously experienced. The number of training programs has increased by 27% over the past 10 years; however, the number of first-year positions has only increased by 15%. Clinical fellow supervision has increased significantly over the years, likely as a result of changes in reimbursement. In 1990, 75% of fellows worked unsupervised in the emergency department versus 23% of first-year fellows, 56% of second-year fellows, and 74% of third-year fellows in the year 2000. The structure of the fellowship curriculum has become more standardized during the past 10 years, with numerous core rotations required by most programs. The percentage of programs offering protected research time has changed significantly over the years, with the amount of time increasing from 40% in 1988, to 95% in 1990, to 100% in 2000. The amount of clinical time has also increased with the transition to a 3-year program.CONCLUSIONS Pediatric emergency medicine continues to expand as a pediatric subspecialty but at a slower rate. During the previous decade, fellowship training has become more structured, with greater emphasis being placed on fellow supervision, standardization of education, and research. These data are meant to assist new as well as established fellowship programs with the development of their training curriculum. PMID- 12592109 TI - Rhabdomyolysis associated with unintentional antihistamine overdose in a child. PMID- 12592110 TI - Traumatic epiglottitis. PMID- 12592111 TI - Acute hematogenous osteomyelitis of the pelvis in childhood: Diagnostic clues and pitfalls. AB - Acute hematogenous osteomyelitis (AHOM) of the pelvis is a rare form of childhood osteomyelitis. Prompted by a recent case, we reviewed the 146 reported cases of pelvic AHOM published since 1966. Classical childhood AHOM of tubular bones usually occurs in older children (mean age, 8.1 y) as opposed to younger children (aged 2-5 y). It is more common in boys than in girls (male to female ratio = 1.5:1). The most common site is the ilium (40%), followed by the ischium (28%) and the pubis (15%). In contrast to AHOM of the long bones, trauma is an uncommon antecedent event in pelvic AHOM. The pain in pelvic AHOM may be referred to the hip, thigh, or abdomen, often leading to misdiagnosis. On average, the correct diagnosis is delayed for 12 days. Such delays have resulted in a permanent disability in 3.4% of the cases. If diagnosed and treated promptly, uneventful recovery can be anticipated in all patients. This case history and review of the literature may facilitate early recognition of pelvic AHOM by primary care physicians, as well as by pediatric or orthopedic specialists. PMID- 12592112 TI - Pediatric nonenvironmental hypothermia presenting to the emergency department: Episodic spontaneous hypothermia with hyperhidrosis. AB - Cases of pediatric nonenvironmental hypothermia are uncommon. When presenting to the emergency department, these patients are often evaluated for possible sepsis/shock, brain tumors, endocrine disorders, and drug ingestions. We report a case of a 5-year-old girl who presented to the pediatric emergency department on two occasions with hypothermia and lethargy. She was found to have an unusual cause of her symptoms: episodic spontaneous hypothermia with hyperhidrosis. PMID- 12592113 TI - Pediatric pericardial tamponade presenting as altered mental status. AB - The purpose of this case report is to illustrate the diagnostic difficulties of pericardial tamponade and to suggest that focused cardiac ultrasound be included in the resuscitative care of pediatric shock. Three cases of cardiac tamponade are presented. Each patient had a syncopal episode and presented with altered mental status and hypotension. Muffled heart tones, distended neck veins, and electrocardiogram and chest radiograph abnormalities were not present. Hypotension was not responsive to intravenous volume expansion treatment. Diagnostic delays would have been prevented if focused cardiac ultrasound had been included in the resuscitative care of shock. PMID- 12592114 TI - Seizures in a 13-year-old girl. PMID- 12592116 TI - Pediatric emergency medicine: Legal briefs. PMID- 12592115 TI - Boundaries of the emergency department. PMID- 12592117 TI - Diagnosis and management of pediatric conjunctivitis. PMID- 12592118 TI - Swallowed penny. PMID- 12592119 TI - Bias in pediatric brodifacoum exposure data. PMID- 12592120 TI - Hypernatremic dehydration and death in an infant. PMID- 12592121 TI - F-18 FDG positron emission tomography and benign fractures. AB - PURPOSE: F-18 2-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) has been used extensively in the imaging of cancer, including metastatic skeletal disease. Although uptake into benign osseous disease has been reported, there is very limited information regarding uptake into benign fractures. This report provides additional information regarding the appearance of benign fractures on FDG-PET images. MATERIALS AND METHODS: Four case reports of FDG-PET scanning are presented in patients with proved benign fractures. RESULTS: In three of these cases, FDG uptake was noted in fractures when images were obtained 17 days to 8 weeks after injury, with the most avid uptake observed when FDG-PET imaging was performed 17 days after fracture. In the patient in whom imaging was performed 8 weeks after fracture, no uptake of FDG was seen in a benign fracture. CONCLUSIONS: Fractures may accumulate FDG to varying degrees, and false-positive findings may occur when FDG-PET imaging is performed to assess for metastases, although the different pattern of uptake and clinical correlation usually allows accurate differentiation of fracture from skeletal metastases. PMID- 12592122 TI - Unsuspected skeletal sarcoidosis mimicking metastatic disease on FDG positron emission tomography and bone scintigraphy. AB - Sarcoidosis is a disease process of unknown cause that exhibits heterogeneous clinical manifestations. Although the mediastinum and lungs are most commonly involved, the granulomatous process may also affect the skeleton, skin, eyes, kidneys, spleen, and liver. A 51-year-old woman with low back pain and no history of cancer was found to have a suspicious lesion at L4 on magnetic resonance imaging. Findings of bone scintigraphy were consistent with multiple metastases, and computed tomography of the chest identified two small indeterminate pulmonary nodules. Similarly, findings of fluorodeoxyglucose positron emission tomography were consistent with widespread skeletal metastases, but a primary tumor was not identified. Biopsy of a left femur lesion was diagnostic of active sarcoidosis with no evidence of cancer. PMID- 12592123 TI - Tc-99m MIBI parathyroid scintigraphy and intact parathyroid hormone levels in hyperparathyroidism. AB - Tc-99m MIBI has been widely used to evaluate hyperparathyroidism based on increased tracer uptake in hyperfunctioning parathyroid tissue. The functional status measurement of parathyroid glands with intact parathyroid hormone (iPTH) levels is also one of the most important diagnostic studies in this disorder. The aim of the current study was to assess the relation between MIBI imaging and iPTH levels. The authors retrospectively reviewed the records of patients with hyperparathyroidism who were referred to their department for Tc-99m MIBI scintigraphy. Sixty-five patients (24 primary and 41 secondary hyperparathyroidism) were included. The iPTH levels ranged from 66.06 to 2,836 pg/ml (normal, 10 to 55 pg/ml). Forty-two patients were MIBI positive and 23 were negative. The iPTH level in the MIBI-positive group was significantly greater than in the negative group in the primary (548 +/- 478 versus 124 +/- 45; = 0.002), secondary (1,155 +/- 692 versus 501 +/- 352; < 0.001), and overall (909 +/- 678 versus 386 +/- 341; < 0.001) groups. For the primary hyperparathyroidism group, 17 of the 24 patients were MIBI positive (71%). When iPTH levels exceeded 200 pg/ml (100%), the diagnostic sensitivity reached 100%. For the secondary hyperparathyroidism group, 25 of 41 patients (61%) were MIBI positive; 24 of 38 patients (63%) had an iPTH level greater than 200 pg/ml, 21 of 27 patients (78%) had an iPTH level greater than 500 pg/ml, and 11 of 12 patients (92%) had an iPTH value greater than 1,000 pg/ml. Tc-99m MIBI parathyroid scintigraphy showed a good correlation with iPTH level for both primary and secondary hyperparathyroidism. Visualization of hyperfunctioning parathyroid glands on Tc 99m MIBI parathyroid scintigraphy was more likely with a higher serum iPTH level in a dose-dependent manner. PMID- 12592124 TI - Efficiency of gamma probe and dual-phase Tc-99m sestamibi scintigraphy in surgery for patients with primary hyperparathyroidism. AB - PURPOSE: The purpose of this study was to determine the value of the intraoperative gamma probe and the efficacy of dual-phase Tc-99m sestamibi imaging in patients with primary hyperparathyroidism. METHODS: Twenty-one patients with primary hyperparathyroidism were examined prospectively. Results of same-day dual-phase Tc-99m sestamibi scintigraphy and intraoperative gamma probe evaluations were compared with the intraoperative findings and histopathologic diagnoses. A 15-mm handheld gamma probe was used to measure gamma activity in the neck and upper mediastinum. Nuclear mapping by gamma probe showed a single quadrant of neck that emitted gamma radiation significantly greater than the other three quadrants, which correlated with the sestamibi scan. RESULTS: Dual phase Tc-99m sestamibi scintigraphy determined and localized parathyroid lesions in 20 patients (sensitivity, 94%). Of the 20 parathyroid lesions removed, 15 were located in normal positions, whereas five were explored in ectopic sites (one within the thyroid, one in the anterior mediastinum, one in a retrotracheal position, one in the carotid sheath, and one in the retroesophageal region). Although the index of thyroid nodules varied from 15.8% to 22.9%, the index for parathyroid lesions was 77.3% to 112.8%. CONCLUSIONS: These results confirm that parathyroid lesions, especially at ectopic sites, can be treated successfully in shorter operative times with minimal complications with the help of the intraoperative gamma probe. PMID- 12592127 TI - Radionuclide imaging of rare congenital renal fusion anomalies. AB - Demonstration of a congenital renal anomaly plays an important role in the treatment of patients with renal infection. These patients are prone to infections because of coexisting urinary tract anomalies such as duplicated ureter, ureter opening anomalies, and urinary stasis. Assessment of renal parenchymal damage resulting from acute or chronic renal infection is the primary indication for radionuclide imaging with Tc-99m DMSA. In addition, this technique allows congenital anomalies to be identified. The authors review congenital renal fusion anomalies identified in children through Tc-99m DMSA imaging. They conclude that Tc-99m DMSA imaging can reveal important diagnostic information about various congenital anomalies, including fusion anomalies. PMID- 12592125 TI - FDG positron emission tomography in the surveillance of hepatic tumors treated with radiofrequency ablation. AB - PURPOSE: Radiofrequency thermal ablation (RFA) is an emerging technique in the treatment of focal hepatic tumors. Magnetic resonance imaging (MRI) and computed tomography (CT) are currently used to monitor hepatic tumors after RFA for residual disease and recurrence. Fluorodeoxyglucose (FDG) positron emission tomography (PET) is an excellent imaging method for the detection of liver metastases, but it has not been thoroughly evaluated as an alternative to anatomic imaging in the surveillance of liver tumors treated with RFA. The purpose of this investigation was to determine the role of FDG-PET imaging in the surveillance of liver tumors treated with RFA. METHODS: Thirteen patients with histories of malignant tumors of the liver treated with RFA and who had received post-treatment FDG-PET scans were assessed retrospectively. One patient had two post-RFA FDG-PET scans, eight patients had concurrent MRI scans, and six patients had concurrent CT scans. Imaging findings were compared with the results of clinical follow-up. RESULTS: There were either recurrent tumors at the ablation site (8 patients) or new metastases (3 patients) in 11 patients. FDG-PET identified all 11 cases and did not misidentify any cases. Of the seven patients with positive PET findings who received an MRI scan, three were also positive on MRI (42.9%); the other four cases were either negative or equivocal. Of the four patients with positive PET findings who received a CT scan, only two had positive CT scan findings (50%). All recurrences diagnosed by PET were confirmed on clinical follow-up. CONCLUSION: In this preliminary study, FDG-PET was superior to anatomic imaging in the surveillance of patients treated with RFA for malignant hepatic tumors. PMID- 12592126 TI - Power Doppler sonography versus Tc-99m DMSA scintigraphy for diagnosing acute pyelonephritis in children: are these two methods comparable? AB - PURPOSE This study assessed the role of renal power Doppler ultrasonography (PDU) to identify acute pyelonephritis (APN) and to determine whether PDU can replace Tc-99m DMSA renal scintigraphy in the diagnosis of APN in children. METHODS A prospective study was conducted in 40 infants and young children (78 kidneys were evaluated) with a mean age of 25.9 months (range, 1 to 68 months) who were hospitalized with a first episode of high fever and bacteruria, possibly APN. All children were examined by PDU and Tc-99m DMSA within the first 3 days after admission. Patients with congenital abnormalities, hydronephrosis, and urinary reflux were excluded. RESULTS Twenty-seven of the 78 kidneys appeared abnormal on Tc-99m DMSA, and 20 of them were abnormal on PDU. Fifty-one of 78 kidneys were normal on Tc-99m DMSA, and 3 of 51 appeared diseased on PDU. The accuracy of PDU was 87%, sensitivity was 74%, and specificity was 94%. The positive predictive and negative predictive values were both 87%. When considering the numbers of lesions in 27 kidneys with positive Tc-99m DMSA studies (38 lesions), PDU did not disclose 16 lesions (false-negative results). Thus, the sensitivity of PDU for diagnosing lesions of APN decreased to 58%. CONCLUSIONS A positive PDU finding should obviate the use of Tc-99m DMSA in patients thought to have possible APN. However, because of a large number of false-negative results (26%) and underestimation of the number of pyelonephritic lesions (low sensitivity of 58%), PDU cannot replace Tc-99m DMSA in the diagnosis of APN in children. PMID- 12592128 TI - In-111 DTPA-octreotide scintigraphy for disease detection in metastatic thyroid cancer: comparison with F-18 FDG positron emission tomography and extensive conventional radiographic imaging. AB - PURPOSE: The utility of In-111 DTPA octreotide scintigraphy (SRS) for disease detection in patients with metastatic thyroid carcinoma (TCA) remains controversial. The authors compared the sensitivity of In-111-based SRS, F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET), and extensive conventional radiographic imaging (CRI) in this type of cancer. METHODS: SRS, FDG PET, and CRI were performed concurrently in 21 patients (age, 56.4 +/- 12.9 years) who had aggressive TCA. Concordance rates % of lesion positivity among pairs of different techniques (A and B) were calculated as the ratio of the number of lesions positive with both techniques divided by the sum of the total number of lesions positive with technique A + total number of lesions positive with technique B, which was then multiplied by 200. RESULTS: The combined use of CRI, FDG PET, and SRS resulted in the detection of 105 lesions, presumed to be due to metastatic deposits. Sensitivities for SRS and FDG-PET imaging were 49.5% and 67.6%, respectively. The lesion detection concordance rates were as follows: CRI versus FDG PET, 80.8%; CRI versus SRS, 74.2%; and FDG-PET versus SRS, 58.6%. Importantly, SRS detected five unexpected lesions, which were negative by both CRI and FDG-PET imaging. In two representative patients, a positive correlation (Spearman's rank = 0.71; = 0.0576) existed between the percentage of lesional In 111 DTPA octreotide uptake and the standard uptake value in eight concordant lesions. CONCLUSION: Although SRS has only moderate sensitivity for disease detection in metastatic TCA, sometimes it can reveal lesions that otherwise would be undetectable by either CRI or FDG-PET imaging. PMID- 12592129 TI - FDG-positron emission tomographic imaging in carcinoma of the breast: interference by massive sarcoidosis. PMID- 12592130 TI - FDG uptake by tongue and muscles of mastication reflecting increased metabolic activity of muscles after chewing gum. PMID- 12592131 TI - Detection of saccular coronary artery aneurysm associated with coronary-pulmonary artery fistula using Tc-99m HSAD scintigraphy. PMID- 12592132 TI - U shaped sternum: a rare malformation demonstrated on bone scintigraphy in an adult patient. PMID- 12592133 TI - Intra-abdominal splenosis mimicking massive gastrointestinal bleeding. PMID- 12592134 TI - Unilateral vesicoureteral reflux detected by diuretic renography without voiding. PMID- 12592135 TI - Incidental detection of skeletal uptake on tetrofosmin cardiac imaging in a patient with multiple myeloma. PMID- 12592136 TI - Discrepancy between Tc-99m HMDP bone scan and F-18 FDG positron emission tomographic images in a patient with small cell lung cancer. PMID- 12592137 TI - Invasive aspergillosis mimicking stage IIIA non-small-cell lung cancer on FDG positron emission tomography. PMID- 12592138 TI - Visualization of collaterals in budd-chiari syndrome with Tc-99m MDP bone scintigraphy and Tc-99m HMPAO-labeled leukocyte scintigraphy. PMID- 12592139 TI - Isolated metastatic adrenal involvement with colon cancer and FDG coincidence detection imaging. PMID- 12592140 TI - Delayed tc-99m sestamibi washout in graves' disease. PMID- 12592141 TI - F-18 FDG brain positron emission tomography and Tl-201 early and delayed SPECT in distinguishing atypical cerebral tumor from cerebral infarction. PMID- 12592142 TI - A combined study of Tc-99m Technegas and Xe-133 gas in suspected congenital bronchial atresia. PMID- 12592143 TI - Scintigraphic visualization of traumatic leptomeningeal cyst on bone scan. PMID- 12592144 TI - Scintigraphic and radiographic appearance of osteomas in Gardner's syndrome. PMID- 12592145 TI - FDG uptake in a huge uterine myoma. PMID- 12592146 TI - Multiple insufficiency fractures in a young woman with anorexia nervosa and bulimia. PMID- 12592147 TI - Tl-201 uptake in chronic hypertrophic pachymeningitis. PMID- 12592148 TI - Radionuclide imaging of cardiac hydatid cyst. PMID- 12592150 TI - [Basic sonographic interpretation]. AB - Basic sonographic semiology includes the dots and lines that are the basic constituants of ultrasound images and the different structures that are the sonographic representation of the different tissues (fluid, fat, calcifications and stones, air, parenchyma). Knowledge of these basic interpretative skills is mandatory to the clinical practice of sonography and allows understanding of the different sonographic manifestations of pathologic processes and identification of artifacts and pitfalls. PMID- 12592151 TI - [Nuchal translucency: technical measurement and value]. AB - Nuchal translucency measurement is a very powerful screening test to detect chromosomal anomalies or other malformations. The technique for measurement is based on strict guidelines that will be described in this paper. Training is mandatory. Risk calculation for chromosomal anomalies (trisomy 21) must also consider maternal age and gestational age. In France, in 1999 the predictive positive value of nuchal translucency for chromosomal anomalies was about 1/7. Other ultrasound markers such as the absence of nasal bone combined with nuchal translucency increase detection. A higher performance will be achieved when ultrasound will be combined with first-trimester maternal serum markers. PMID- 12592152 TI - [Ultrasonography of twin pregnancies]. AB - Sonographers must be familiar with the diagnostic tools available to them for determining twin pregnancy type. During the first trimester, these tools include the number of chorionic sacs, the number of yolk sacs, and the presence of an inter-twin membrane. During the second trimester, sonographers must look for a difference in sex, the presence of two separate placentas, and a thick vs. thin membrane, or the absence of a membrane. Complications are frequent in twin pregnancies and may be maternal or fetal. Measuring cervical length is the only way for sonographers to predict pre-term labour. A cervical length of< or =2.5 cm before 23 weeks is significant. Some fetal anomalies are more frequent in twin pregnancy than in single pregnancy. The more frequent are: esophageal atresia, omphalocele and cardiac anomalies. Some fetal anomalies are peculiar to twin pregnancy: Siamese twins, digestive atresia, craniofacial deformation and club foot. A significant growth asymmetry is defined as a CRL of 3 or more millimetres and a difference of 20% in weight. The poly-oligo sequence represents a severe level asymmetry of amniotic fluid in monodiamniotic twins where survival rate without sequelae is 13.6%. The twin-twin transfusion syndrome is an event occurring after the apparition of the poly-oligo sequence, which manifests by discordances in weight and hemodynamic changes leading to myocardial repercussion. Twin embolization syndrome follows the in utero death of a co-twin in a monochorionic twin pregnancy; ischemic events must be monitored in such cases. Cord entanglement is unpredictable and is specific to monochorionic monoamniotic twin pregnancies. Finally, an acardius is a parasitic twin that feeds on its co-twin, resulting in the viable twin's death in 50% of cases. PMID- 12592153 TI - [The fetal head: from screening to diagnosis]. AB - The implementation of a prenatal screening ultrasound program to the general population requires a few rules. The sonographers must be adequately trained and the required imaging planes of the fetal head must be easy to acquire and be readily reproducible irrespective of the technology used. Screening for the presence of a potential local or global anomaly can be mainly established from a reference image. The diagnosis of an anomaly is confirmed from a different medical approach and is usually done during the second trimester (21 weeks GA). PMID- 12592154 TI - [Doppler monitoring of fetal circulation from multiple arteries over several days to improve evaluation of fetal prognosis]. AB - Intrauterine growth retardation and fetal hypoxia are currently related to placental insufficiency. Fetal biometry assessed by echography is entirely adapted to follow the growth and integrity of the principal fetal organs. Hypoxia induces an hemodynamic adaptation which can be detected and quantified by Doppler. The objective of this article is to review the evolution of the fetal Doppler practice for the last 20 years and especially to show that isolated Doppler measurement and only from one site (umbilical or cerebral or aortic) have a moderate negative predictive value of fetal outcome, compared to the study of the fetal hemodynamic evolution (degradation) from several sites and during several days. We will insist on the fact that (a) umbilical Doppler only gives information on placental blood flow and this information does not reflect neither the adaptation to hypoxia nor the consequences of this adaptation, (b) cerebral Doppler accounts for the vascular response to the pO(2) reduction but it does not allow to predict the consequences of this response, (c) the simultaneous study of the placental hemodynamic time course degradation and the cerebral vascular response to hypoxia allows quantification of the cumulative deficit of fetal oxygenation during this period and evaluation the adverse consequences of a sustained flow redistribution toward the brain. Finally, if cerebral vasodilation in response to hypoxia can be considered as a physiological compensatory mechanism, it is associated after several days to the appearance of irreversible fonctional (abnormal fetal heart rate) or organic (cerebral lesions) abnormalities. Adverse effects of this process are illustrated during episodes of acute hypoxia (malaria crisis of several days) or during sustained exposure of the fetus to hypoxia (pregnancy-induced hypertension). PMID- 12592155 TI - [Acute appendicitis: to image or not to image?]. AB - Acute appendicitis is frequently clinically suspected. However, about 50% of emergency room patients with such a diagnosis do not have acute appendicitis and between 20-25% of patients undergoing appendectomy based on clinical diagnosis have a normal appendix. On the other hand, if left untreated acute appendicitis may result in peritonitis. The purpose of this article is to review the indications for imaging patients with clinical suspicion of acute appendicitis, to describe the US and CT features of acute appendicitis, to review the advantages and limitations of US and CT, and to present the differential diagnosis to be considered in patients with right lower quadrant pain. PMID- 12592156 TI - [Smoking and Pregnancy. Introduction]. PMID- 12592157 TI - [Smoking, fertility and very preterm birth]. AB - In France, many couples in procreating age continue to smoke, despite recommendations to stop smoking before or during pregnancy. We reviewed the epidemiological associations between maternal or paternal smoking and both infertility and very preterm birth. It appears clearly that maternal smoking impairs natural and assisted fecundity. Furthermore, maternal smoking enhances the rate of ectopic pregnancies and spontaneous abortions. The role of maternal smoking on very preterm birth before 33 weeks of completed gestation appears in recent population based-study or case-control studies that include several hundred very preterm neonates. But the role of smoking on very preterm birth among hypertensive mothers and primiparae is still debated. The detrimental effect of smoking on fecundity appears to be reversible when the mother and father stop smoking. The real effect of interventions promoting cessation of smoking during pregnancy in reducing the rate of very preterm births remains to be studied. PMID- 12592158 TI - [Smoking, fetal pulmonary development and lung disease in children]. AB - Prenatal exposure to tobacco smoke is associated with a significant increase in respiratory symptoms and a decrease in lung function in children. Epidemiological studies have demonstrated that tobacco smoke exposure represents one of the most important respiratory risk factors in childhood. Nicotine crosses the placenta and activates the nicotinic receptors which are present in a wide range of lung cells during the prenatal period. In utero tobacco exposure is responsible for an increase in alveolar volumes and a decrease in airway walls caliber, which explains the decrease in airflow that is observed during the first days of life in infants of smoking mothers. These structural and functional disturbances explain why these infants are more susceptible to respiratory irritants. The alterations in ventilatory control induced by prenatal exposure to nicotine explain the link between tobacco smoke exposure and the sudden infant death syndrome. This underlines the importance of adequate information and preventative measures to decrease this major and avoidable cause of respiratory morbidity and mortality in infants. PMID- 12592159 TI - [Prenatal exposure to tobacco smoke and cognitive development: epidemiology studies]. AB - Studies concerning smoking during pregnancy and cognitive development of the child were reviewed. Some studies tend to support the existence of an association between smoking during pregnancy and a less optimal cognitive development of the child. The meaning of these relationships remains obscure, since it cannot be assumed that they are due to maternal cigarette smoking: they could be due to different characteristics of smokers or non-smokers. In the older studies, potential confounding factors were not adequately taken into account and in some recent studies, the observed difference was no longer significant after taking into account the maternal educational level and the family stimulation of the child. Behavior problems are associated with smoking during pregnancy in nearly all studies but the same problem of confounding factors remains. It is thus difficult to conclude presently from these studies if there is a causal relationship between prenatal exposure to tobacco smoke and cognitive development. PMID- 12592160 TI - [Mechanisms of action of tobacco smoke on the developing brain]. AB - Maternal smoking is associated with an increased risk of intra-uterine growth retardation and conduct disorder in their children, and is a major risk factor for sudden infant death syndrome. Among the numerous compounds present in cigarette smoke, carbon monoxide and nicotine have been largely studied. They can both affect the fetal brain by inducing intra-uterine hypoxia or by acting directly on the developing brain. PMID- 12592161 TI - [Smoking and sudden infant death syndrome]. AB - Assessment of the causation relationship between two phenomena requires the demonstration of an epidemiological association, a temporal and asymmetric sequence, and a biological gradient and identification of the biological mechanism(s). All epidemiological studies on sudden infant death syndrome (SIDS) and smoking have encountered major bias and difficult data interpretation but they all have estimated that maternal smoking caused a 2 to 3-fold increased risk of SIDS. Nicotine may interact with non-neuronal nicotinic receptors in the lung, peripheral nicotinic cholinergic and adrenergic chemoreceptors, and brainstem nuclei and has been largely studied. More accurate knowledge concerning the biochemistry and specific features of nicotinic receptors will be useful to explain the way nicotine alters breathing at rest and during hypoxia. Uncertainty about the casual relationship in no way means the fight against smoking is not warranted. PMID- 12592162 TI - [How can we help pregnant women stop smoking?]. AB - Smoking is a behavior maintained and enhanced by nicotine-induced dependence. Despite awareness and knowledge of the associated health risks many smokers find it considerably difficult to quit. Nicotine withdrawal symptoms and undesirable effects such as depression and weight gain serve as justification for the numerous unsuccessful attempts in smoking cessation. Yet, we have now come to the end of empiricism. Treatments that have been shown to work exist and international evidence-based recommendations for cessation interventions have been established: brief advice, nicotine replacement therapy and behavioral and cognitive therapies. Measuring nicotine dependence using the Fagerstrom test helps to define the therapeutic strategy. Blood cotinine level can be measured during pregnancy. It is obvious that therapies can only work for smokers who are motivated to stop smoking. Before reaching the decision to quit, the smoker goes through a process during the course of which the role of information and advice from health professionals are paramount. When they became pregnant, young women are not necessary ready to quit. For many of them, pregnancy is sufficient motivation enabling them to stop smoking, but for thirty percent of them, probably the most dependent, it would be very difficult to stop without a specific program of help. PMID- 12592163 TI - [New non-invasive technique for prenatal diagnosis: analysis of maternal blood]. AB - The presence of free fetal DNA circulating in the maternal plasma or serum has been demonstrated by Lo et al. in 1997. In 2002, fetal DNA has been used as a tool for prenatal diagnosis. The main indications for this technique are currently limited to determination of fetal sex for X-linked diseases as well as determination of fetal RhD in RhD negative women. Other more exceptional applications are also possible, including detection of translocations or search for de novo point mutations but currently remain in the research phase. PMID- 12592164 TI - [Routine cytomegalovirus screening during pregnancy: is it worthwhile?]. AB - The issue of routine antenatal screening for cytomegalovirus (CMV) infection raises a controversy in France. Supporters of a screening policy argue that this frequent congenital infection (about 1% of all newborns) is a known cause of neurological impairment or hearing loss, which could be prevented by the prenatal screening of the infection. However the majority of infected fetuses will be free of any symptom, and there is currently no reliable prognostic marker in the antenatal period. Moreover, no prenatal treatment has proven to be effective and safe. A screening strategy might lead to iatrogenic concerns, including parental anxiety, invasive prenatal testing, and terminations of pregnancies for unaffected fetuses. Based on current knowledge, routine antenatal screening of CMV infection during pregnancy cannot be recommended. Further research should focus on the development of a safe and effective prenatal treatment, and the development of better diagnostic tests. PMID- 12592165 TI - [National program for neonatal screening for cystic fibrosis: implementation and preliminary results]. AB - Neonatal screening for cystic fibrosis was decided by the national medical authorities after a common investigation conducted by the French association ADPHE and national health insurance fund. Based on therapeutic progress and the proposed method using determination of blood immunoreactive trypsin then study of the main CF mutations, there is strong hope of effective CF detection and clinical benefit for the patients. PMID- 12592166 TI - [Neonatal detection of sickle cell disease]. AB - An experimental program for neonatal detection of sickle cell disease (SCD) was performed in France in 1990. Our data indicated a high prevalence of SCD, one patient in 1,250 newborns tested. The French national program for neonatal screening for SCD was set up in 1995 by the Association Francaise pour le Depistage et la Prevention des Handicaps de l'Enfant (AFDPHE). This is not a systematic program, but only targeted at newborns at risk for SCD. PMID- 12592168 TI - [Acid base balance in the fetus during labor: pathophysiology and exploration methods]. AB - Although the majority (about 90%) of cerebral palsies are related to antenatal or postnatal causes, search for acute fetal distress during labor remains one of the primary objectives of obstetrical surveillance. Acute fetal distress leads to hypoxemia which induced hypoxia and then asphyxia of most of the organs, including the noble organs such as the brain. Most of the parameters used for fetal monitoring have good sensitivity but low specificity. Furthermore, there is a weak correlation between the different parameters measured and the neonatal status which remains difficult to predict with certainty. The general "philosophy" behind fetal monitoring consists in detecting warning signs and, when warning signs are observed, to measure one or more other parameters to assess the state of hypoxia and its consequences. Fetal heart rate is a warning sign used by all teams. Irrespective of the method used to analyze fetal heart rate when anomalies are detected, metabolic acidosis induced by anoxia is an indispensable element for assessing severity. The development of fetal hypoxia is related to two essential phenomena: anaerobic glycolysis with lactate production and glycogenolysis to meet energy needs. Metabolic acidosis can be measured directly by scalp pH or lactate measurements, or indirectly by measuring its effect on heart conduction as expressed in the ST segment. The underlying physiological mechanisms and the methods used to evaluate acid base balance are described here. PMID- 12592169 TI - [Intrapartum fetal monitoring: old or new techniques?]. AB - The objective of intrapartum fetal monitoring is to prevent fetal asphyxia and fetal death during labor. Although a large proportion of neurologic sequelae are of antepartum origin, criteria have been defined to relate cerebral palsy to an intrapartum event, including fetal asphyxia. Since the introduction of fetal heart rate monitoring, no changes in the cerebral palsy rate have been observed. Several other methods of fetal surveillance have been more or less recently developed: fetal scalp blood pH and lactates, fetal pulse oximetry and fetal electrocardiogram. The benefits and limits of these new tools are analyzed in this review. PMID- 12592170 TI - [Neurological prognosis of term infants with perinatal asphyxia]. AB - Neonatal encephalopathies following birth asphyxia are the first features of cerebral insult. They never miss when asphyxia is directly involved in cerebral impairment. Mild encephalopathies have constantly a good prognosis. Conversely, moderate and severe encephalopathies are associated with poor outcome (death or severe handicap) in 25% to 100% of cases. Prognosis of these moderate and severe encephalopathies can be assessed during the first ten days of life by 3 complementary ways: clinical exam, electrophysiology and imaging. The most information is obtained from the EEG and MRI which together nearly reach 100% for both predictive positive and negative values for severe neurological sequelae. PMID- 12592171 TI - [Orientation after peripartum asphyxia in the maternity ward: which infants should be transferred to pediatric care units?]. AB - Per-partum anoxia is a frequent situation facing the pediatrician in the maternity ward. The question is to decide which infants require care in a specialized unit. If transfer is decided, the infant must be referred to an appropriate pediatric unit (intensive care or neonatal unit). Cases of severe anoxia are exceptional. Intermediary situations are however much more frequent and raise difficult evaluation problems due to the lack of any specific test. The pediatrician must rely on a combination of elements from the clinical presentation, the medical history, the clinical course, and laboratory tests. Different elements suggest a prudent approach with referral to a pediatric unit. These elements include: imperfect clinical recovery (5-min Agpar <7), major intensive care at delivery (intubation, ventilation, vasoactive agents), anomalies in the cord blood or first hour blood tests (cord pH<7, base deficit 12, cord or blood lactate 9 mmol/l). Obstetrical circumstances which led to per partum anoxia must be well identified because those interrupting placental flow (abruptio placenta, uterine rupture) suggest prudence is necessary even if the infant appears to have recovered well. All neonatal disorders (macrosomia, prematurity, infection, respiratory distress) increase the risk of rapid decompensation and may argue for hospitalization. Likewise, if even minimal signs of neurological, respiratory or hemodynamic disorders are present from birth to two hours, surveillance in a specialized unit is required, the level depending on local facilities. Certain situations nevertheless always require referral to a pediatric intensive care unit: use of vasoactive drugs, respiratory distress, abnormal neurological exam, poor recovery (5-min Agpar <4).Finally, it must be remembered that per-partum anoxia is rarely predictable and can occur any at any time of day or night. The pediatrician must also train other delivery room personnel, including the midwives, in intensive care techniques. PMID- 12592172 TI - [Newborn asphyxia at term during delivery]. AB - Recruitment, work load and morbidity linked to newborn asphyxia during delivery at term: a study from Pediatric Mobile Intensive Care Units. OBJECTIVE: In a population of term neonates transported by the mobile intensive care units (MICU), we aimed to determine the incidence of neonates with anoxic-ischemic encephalopathy related to asphyxia, to analyze in this population the difficulties of management, and to try to identify which of these newborns require new therapeutic strategies. METHODS: This retrospective study was performed over a 2-year period (2000 and 2001) in 3 paediatric MICU from the Ile de France area. During this period, 7,648 infants were transported including 3,301 newborns of more than 36 weeks of gestational age and less than 72 hours of life. These neonates came from 73 different hospitals. Among these 3,301 infants, 237 neonates (124 boys and 113 girls) with anoxic-ischemic encephalopathy related to asphyxia were selected in the present study. Inclusion criteria were association of one obligatory criterion of fetal distress during delivery and at least one criterion of neonatal asphyxia or one criterion of anoxic-ischemic encephalopathy. Data were compiled and analyzed with Epidata package and Epi info package, respectively. RESULTS: These 237 neonates with anoxic-ischemic represented 12% of MICU activity at the same gestational and postnatal ages. The mean gestational age was 39.5 + 1.5 weeks. The mean birth weight was 3,188 + 559 g. More than 50% of these neonates were born in level I maternities. Fifty-three percent of the infants were born by caesarean section. Eighty-three percent of the neonates had an Apgar score at 1 minute <3. Eighty-eight percent of the neonates received resuscitation care at 5 minutes of life and 34% of these had an Apgar score at 10 minutes <5. In 50% of the cases, the MICUs arrived at the maternity of delivery within 1 h 45 min of life and transportation of the neonates was completed after 3 hours of life. The neonates were transported to an intensive care unit in 88% of the cases (half to a polyvalent intensive care unit and half to a neonatal intensive care unit). Forty-four percent of transported neonates had no encephalopathy, 30% had a severe encephalopathy or seizures, 27% had multiple organ failure. Mortality reached 28% and encephalopathy accounted for two thirds of these deaths. Neonates who arrived in pediatric care units after 3 hours of life had more severe morbidity than neonates who arrived before 3 hours of life. CONCLUSION: Pediatric MICUs transport the most severely affected neonates. The initial clinical state is critical, and systemic and neurological complications are frequent and severe. Calls to the MICU should be made earlier in order to enable a better impact of new neuroprotective strategies. PMID- 12592173 TI - [Organization of patient management in level II centers in the Paris area: a prospective survey]. AB - Perinatal asphyxia is a common emergency for both obstetricians and pediatricians. A prospective study was conducted in 14 maternity hospitals (type II centres) in the Paris suburbs in order to assess pediatric activity and neonatal morbidity associated with supposed perinatal asphyxia in term newborns. Pediatricians were called in at birth very frequently: 1/20 deliveries. Intubation and/or resuscitation procedures were needed in 20% of cases and 20% of infants were referred to a neonatal unit for birth asphyxia or associated pathology. Moderate encephalopathy was observed in 1.5% of all term newborns who needed medical intervention for supposed birth asphyxia. PMID- 12592174 TI - [Per-partum anoxia and handicaps: epidemiological aspects]. AB - Although per-partum anoxia accounts for only a small part (less than 10%) of the etiology of cerebral palsy, statistical computations, based on current epidemiological data, show that, when per-partum anoxia is associated with suboptimal care 9 out of 10 cases of cerebral palsy could be attributed to the anoxia. PMID- 12592175 TI - [Medical-legal aspects: the obstetrician as a defendant or as an expert]. AB - Most claims ab out term intrapartum fetal asphyxia (or cerebral palsy presumed to be in relation with it) are based on abnormal intrapartum fetal heart rate patterns, and presumed excessive delay from decision-to-delivery and/or inadequate care. For the obstetrician, whether as a defendant or a judicial expert, the difficulty lies in data interpretation which should not be reduced to an analysis of fetal monitoring, but should consider the context, the explorations used to be sure of abnormal fetal heart rate interpretation, and appropriate delay for delivery in light of the findings. PMID- 12592176 TI - [Medical-legal aspects: the pediatrician as a defendant or as an expert]. AB - Claims concerning management of term intrapartum fetal asphyxia account for nearly 45% of all claims filed against pediatricians practicing in the perinatal setting. We recall here briefly the legal and judiciary context under French law, calling upon ten years experience as a medical expert to describe the principal circumstances and clinical forms of acute intrapartum fetal distress which have lead to claims. A few precautionary and preventive measures which should be taken for the clinical forms of intrapartum fetal asphyxia which carry the greatest risk of claims are presented. Finally, practical elements which should be implemented are proposed if a claim is filled against a pediatrician. PMID- 12592177 TI - [Management of first trimester miscarriages]. AB - INTRODUCTION: First trimester miscarriages complicating 15 to 20% of pregnancies, are the leading cause of gynecology emergencies. OBJECTIVES AND METHODS: This review exhaustively summarized the French and English reports found on Medline data base about the management of first trimester miscarriage. RESULTS: The treatment of reference remains surgical evacuation. It is generally used to avoid haemorrhage and intrauterine infection but is also required for women who wish an immediate termination of pregnancy. Recently, alternatives have been proposed, such as expectant management or medical treatment by misoprostol, to improve patient satisfaction and to reduce complications and costs generated by surgery. Expectant management allows to avoid curettage in most of incomplete miscarriages without any increase of the rate of complications or bleeding duration. Vaginal misoprostol allows complete expulsion in 70 to 90% of missed pregnancies without any increase of the rate of complications. CONCLUSION: Today, we can propose to motivated and informed patients an alternative to surgical treatment but large randomized studies still are needed to assess benefits and risks for each method. PMID- 12592178 TI - [Imaging pelvic floor prolapse]. AB - OBJECTIVES: Dynamic MR imaging (MRI) may be used as an alternative to dynamic cystoproctography for the evaluation of pelvic floor prolapse. Our study aimed to define the range of dynamic pelvic MRI aspects observed during the exploration of pelvic prolapse. MATERIAL: Fascial and anatomic defects can include a combinaison of cystocele, rectocele, uterine prolapse, enterocele and vault prolapse. Acute diagnosis of the coexisting abnormalities is essential in planning reconstructive procedures so that the risks of recurrence and reoperation can be minimized. At this time colpocystography is the study of choice to evaluate pelvic floor dysfunction. Dynamic magnetic resonance imaging for exploration of pelvic floor dysfunction is under evaluation. RESULTS: Pubococcygeal line and puborectalis muscle were the references points. The grading system is based on degree of organ prolapse through the hiatus and the degree of pubo-rectalis descent and hiatal enlargement. Unfortunately, there is a real discordance between the reality of the prolapse and its imaging. However, a new reference line, the mid pubic line, was drawn on the magnetic resonance image to correspond to the hymeneal ring marker used in clinical staging. Intra-operative findings were considered the gold standard against which physical examination, dynamic colpocystodefecography and MRI were compared. Using these criteria the sensitivity, specificity and positive predictive value of MRI were 70%, 100%, 100% for cystocele; 42%, 81%, 60% for vaginal vault or uterine prolapse; 100%, 83%, 75% for enterocele; 87%, 72% and 66% for rectocele. CONCLUSION: MRI appears to be an important adjunct in the comprehensive evaluation of the female pelvis. Except for vaginal vault, pelvic prolapse is accurately staged and pelvic organ prolapse reliably detected. The technique is rapid, non invasive and cost effective, it allows the clinician to visualize the whole pelvis using a single dynamic study that provides anatomical details. PMID- 12592179 TI - [Pemphigoid gestationis: a study of 15 cases]. AB - INTRODUCTION: Pemphigoid gestationis is a rare subepidermal bullous dermatosis generally occurring during the 2(nd) or 3(rd) quarter of gestation or in the postpartum period in women who already have been pregnant. OBJECTIVE: The aim of this work is to draw a profile of the epidemiology, clinical aspects, treatment and evolution of the disease by studying hospital series. METHODS: In this retrospective study, 15 cases of pemphigoid gestationis confirmed by direct immunofluorescence, followed in the department of dermatology between 1983 and 1999, were included. RESULTS: The age of onset was 19 to 39 years (mean age: 30 years). In 73% of cases, pemphigoid gestationis occurred in women who had already been pregnant, and appeared during the last 3 months of gestation in 11 patients. In all cases, purities was the first symptom, followed by a erythematous maculopapular eruption. In the steady state of the disease, all patients had annular confluent erythematous papules with herpes, like vesicles predominant in the umbilicus. The diagnosis of pemphigoid gestationis was confirmed by direct immunofluorescence in all cases demonstrated linear staining of C3 at the basement membrane zone. Systemic corticosteroids (0.5-1mg/kg/day) were used in 54% of reported cases. Dapsone was efficient in 26% of patients. 20% of patients were treated with oral antihistamine and topical glucocorticoid. Recurrence occurred in postpartum in 53,3% of patients. Two patients had recurrence during the following pregnancies. CONCLUSION: PG remains a rare dermatitis of pregnancy. Our series is comparable to the literature: the late occurrence of PG during the course of pregnancy, the high frequency of multigravida women, the lack involvement in the newborn, however with some particularities: the frequent involvement of the face and the efficiency of dapsone. PMID- 12592180 TI - [Maternal and fetal prognosis of hypertension and pregnancy in Africa (Senegal)]. AB - OBJECTIVES: The aims of this study were to record the different types of hypertension associated with pregnancy and to assess the incidence of hypertension and its gravity in Senegal. METHODS: Over a two-year period, a cohort of pregnant women with hypertension according to the American working group classification of hypertension and pregnancy, was studied. A group of 47 non hypertensive women were matched for age and parity. Modalities of delivery were studied: maternal death, type of delivery, birth weight. RESULTS: Among 2,400 deliveries, hypertension was observed in 94 women wih, mean age 33 years. The incidence of hypertension was 3.9% and the incidence of preeclampsia was 2.5%. The different types of hypertension were: Type I: 44 (47%), Type II: 16 (17%), Type III: 18 (19%), Type IV: 16 (17%). Echocardiography showed 30 cases of left ventricle hypertrophy with 3 cases of systolic dysfunction. Thirty-five patients had undergone a caesarean. Forty-seven infants had a birth weight below 2,000 g. Maternal mortality was 12.7%, fetal and neonatal mortality was 50%. There was a 21-fold higher chance of caesarean section in hypertensive women (p<4 x 10-4). Neonatal mortality was 36 times higher (p (4 x 10-6) than in the control group with a birth weight lower birth weight 975 g (p<10-6). Women suffering from toxemia gave birth to children having a lower birth weight (-543 g) (p<5.10-3), but, there was no significant difference concerning caesarean (p<7*10-1) maternal, fetal and neonatal mortality (p<9. 10-1) compared with other sub groups. CONCLUSION: In developing countries, hypertension in pregnant women is a severe condition responsible for disease and handicaps which could be avoidable at little cost through a better policy of detection and good quality multidisciplinary management. PMID- 12592181 TI - [The role of amniocentesis in the management of chorioamnionitis with Listeria monocytogene]. AB - Listeriosis is prevalent in pregnant women. Associated morbidity includes miscarriage, chorioamnionitis, intrauterine and neonatal death. Maternal symptoms are not specific and the diagnosis is difficult with a high rate of false negative microbiology results. We report here the case a patient who developed a chorioamnionitis at 31 weeks gestation. Diagnosis was established by examination of the amniotic fluid. We report a case of Literiosis in pregnancy diagnosed by direct examination of amniotic fluid obtained by amniocentesis. PMID- 12592182 TI - [Pelviperitonitis and intestinal occlusion: two unusual cold-knife conisation complications]. AB - Complications associated with cold-knife conization are mainly hemorrhage, genital and urinary infections. We presented two cases of accidental posterior colpotomy with Douglas pouch wound initially unnoticed at the time of the cold knife conization, and secondary complicated by a pelviperitonitis for the first case and by an incarceration of the bowel loop in the vaginal wound for the second case. PMID- 12592183 TI - [Cancer of the vulva and pregnancy: two cases and review of the literature]. AB - The cancer of vulva is predominantly a disease of postmenopausal women. But some cases have been reported in young women during pregnancy. Surgical management during pregnancy depends on several factors: gestational age at the diagnosis, lesion size, depth and location, lymph node status. We have conducted a national survey and report here the only two cases of squamous cell carcinoma diagnosed and treated during pregnancy at 26 and 15 weeks gestation. All vulva symptoms without signs of infectious should be examined with attention in order to detect vulvar lesions. When in doubt, a biopsy must be obtained. Management of vulva carcinoma is not modified during pregnancy, but the age of pregnancy must be considered. PMID- 12592184 TI - [Spontaneous rupture of utero-ovarian vessels in postpartal period: a case report and review of the literature]. AB - Spontaneous rupture of utero-ovarian veins occurring during pregnancy or the peripartal period is generally considered to be a medical curiosity (approximately 100 reported cases) rarely mentioned in obstetric textbooks. It is nevertheless a dramatic cause of maternal and /or fetal mortality. The available statistics are the following: 60% of the cases are directly related to labor and 50% occur in primiparas; when the rupture is associated with labor, mortality is 40%. Occurring outside of labor, mortality rate is approximatively 10%. The perinatal mortality remains high at 30%; in 75% of the cases, the broad ligament is the site of rupture (in many cases, the site of rupture is not found, at laparotomy or autopsy). We report the case of a patient who developed ovarian vein rupture on the 3rd postpartum day. PMID- 12592185 TI - [Intrauterine device appendicitis: an exceptional complication]. AB - We report a case of migration of an intrauterine contraceptive device to the appendix. In this patient with a clinical presentation suggestive of appendicitis, transvaginal ultrasonography visualized a device located outside the uterus, near the small bowel. The device, which was partially embedded in the appendix, was removed via laparoscopy and laparotomy. Appendicular migration of an intrauterine device is exceptional. A literature review spanning the past years only revealed a few reported cases. Most of the authors recommend removal because of the potential for inflammatory reactions that can cause bowel obstruction and perforation. PMID- 12592186 TI - [Guidelines (brief text): management of cutaneo-mucosal herpes in the immunocompetent patient (ocular manifestations excluded)]. PMID- 12592187 TI - [Antenatal prevention of early neonatal bacterial infection (September 2001)]. PMID- 12592188 TI - [About the article "Can beliefs, medicine, and justice get along?" by L. de Ponceville]. PMID- 12592189 TI - [About "Cystocele repair by the vaginal route with a tension- free sub- bladder prosthesis"]. PMID- 12592191 TI - [Female fertility preservation before sterilizing treatment: contribution of ovarian tissue cryopreservation]. AB - Longer survival after anticancer treatment has lead to concern about the long term adverse effects. Altered fertility is of particular importance. Before sterilizing treatment, three non-exclusive methods can be proposed to preserve female fertility: in vitro fertilization followed by cryopreservation of embryos, cryopreservation of mature ovocytes, cryopreservation of ovarian tissue. The method or methods chosen will depend on the age of the patient, here marital status, the urgency of the treatment, and the type of disease. Embryo cryopreservation is a routine practice in medically assisted reproduction centers, while cryopreservation of mature ovocytes and ovarian tissue is still in the experimental phase. It is known however that mature ovocytes can be used after cryopreservation. Cyropreservation of ovarian tissue is a more difficult problem. To date, there have not been any pregnancies or births after freezing thawing of human ovarian tissue. This tissue could be used in two ways: autograft and in vitro folliculo-ovocyte maturation. Despite the uncertainty concerning use, women cryopreservation of ovarian tissue quite well. PMID- 12592192 TI - [The increase in multiple births and its consequences on perinatal health]. AB - OBJECTIVE: To describe the trends in multiple maternity rates and the main responsible factors, and to present the main medical risks associated with multiple births. METHOD: A review of studies describing the situation in Europe and America North. RESULTS: Multiple maternity rates have increased dramatically in France and in other Western countries since the mid 1970s. However the triplet maternity rates leveled off in the 1990s in several European countries. One fourth to one-third of the increase in multiple maternities is explained by the increase in maternal age. Moreover 30 to 50% of twin maternities and at least three-fourths of triplet maternities would occur after fertility treatments, according to surveys or estimations made from various sources. Twins and triplets have high risks for mortality and morbidity. In France in 1995/98, among live birth twins, the proportion of births before 37 weeks was 43.7% and the proportion of births before 33 weeks was 8.1%. The risk fraction of preterm delivery (<37 weeks) attributable to twins rose from 13% in 1981 to 19% in 1995/98. This trend is explained by the increase in occurrence of multiple births as much as by an increase in preterm births among twins. A similar situation is found in Canada and the USA. Whereas triplets have very high risk of preterm birth, the risk fraction attributable to triplets is very much lower than for twins: in Canada it was 1.1% in 1995-97. CONCLUSION: These results suggest that research and preventive interventions should not be restricted to triplets, but should also include twins. PMID- 12592193 TI - [Ovarian pregnancies: revaluation of diagnostic criteria]. AB - OBJECTIVES: To clarify the clinical features of ovarian pregnancy and to show the incapacity of Spielberg's criteria to establish the diagnosis. MATERIAL AND METHOD: and method. Retrospective experience of ovarian pregnancies in a single maternity unit over seven years with comparison between new diagnostic criteria and those of Spiegelberg. RESULTS: Thirteen ovarian pregnancies identified (incidence=1 for 1400 deliveries). History of pelvic disease (one case), use of an IUCD (five cases), pelvic pain (all the cases), metrorrhagia (four cases), hemorrhagic shock (two cases). Diagnosis was evoked only once by ultrasound. Eleven patients were treated by laparoscopy. None of the ovarian pregnancies in the present series met the criteria of Spiegelberg's definition. CONCLUSION: Ovarian pregnancy is a rare form of ectopic pregnancy, but its incidence is certainly underestimated. Search for difficult to detect ultrasonographic features is essential. Criteria, other than those described by Spiegelberg, when present together confirm ovarian pregnancy: serum beta-hCG level > or = 1000 IU/l and uterine vacuity at vaginal ultrasonography; ovarian implication confirmed by surgical exploration, with bleeding, visualisation of chorionic villi or presence of an atypical cyst on the ovary; normal tubes; absence of serum beta-hCG after treatment of the ovary. Ovarian pregnancy does not compromise subsequent fertility of these patients. Recurrence is exceptional. PMID- 12592194 TI - [Neonatal consequences of preterm premature rupture of membrane (PPROM) at 24-34 WG: 118 singleton pregnancies]. AB - OBJECTIVES: To evaluate the outcome of preterm infants born after preterm premature rupture of membranes (PPROM) between 24 and 34 weeks gestation (WG). METHODS: One hundred and eighteen consecutive singleton infants were included in a prospective unicenter study during 3 years. A PPROM management had been instituted by the local obstetrician's board committee. In case of suspected chorioamniotitis defined by maternal or fetal criteria or in case of fetal heart rate abnormalities, a fetal extraction was decided. Diagnosis of chorioamnionitis was confirmed regarding the results of placenta culture and/or histology. Neonatal sepsis was defined by positive blood placenta culture or by the association of 2 positive bacterial cultures with a CRP>15 mg/l. RESULTS: The mean gestational age was 29.8 WG (24-34 SA) for PPROM and 30.4 WG for delivery. Mean latency period between PPROM and delivery was 119 hours +/- 208. The incidence of choriamnionitis was 31%. The overall incidence of neonatal sepsis was 15%. Neonatal mortality was 11.7%. Bronchopulmonary dysplasia occured in 8.4% of the liveborn cases and cranial ultrasound abnormalities (HIV III-IV, kystic LPV) in 11.7% of overall cases. Cerebral lesions were positively correlated with hypotension and negatively correlated with GA. Cerebral lesions seemed to occur more frequently in case of chorioamnionitis but it is not significant. CONCLUSION: In this study, in case of PPROM between 24-34 SA, gestational age and hypotension are correlated with neurologic injury. PMID- 12592195 TI - [Fallopian tube torsion: five cases with no other element]. AB - Isolated Fallopian tube torsion is extremely rare and diagnosis is difficult. Laparoscopy is necessary, but surgery is often performed too late to preserve the tube. Five cases of Fallopian tube torsion with no other element are presented. Diagnostic difficulties, causes, ultrasonographic findings and treatment are discussed. Early diagnosis and surgical intervention is mandatory when exceptional isolated Fallopian tube torsion is suspected particularly in a nulliparous young woman. PMID- 12592197 TI - [Adnexal defect identified with hysterosalpingogram and laparoscopy during infertility evaluation]. AB - OBJECTIVE: To describe a previously unreported adnexal defect as a cause of an abnormal hysterosalpingogram (HSG), identified during infertility evaluation. MATERIAL AND METHOD: An abnormal hysterosalpingogram (HSG) consistent with partial left tubal obstruction or hydrosalpynx was identified during the evaluation of a 33 year-old nulligravida with no prior surgical history. At laparoscopy, the diagnosis was revised when a approximately 3x4cm regular, ovoid, physiologic aperture was noted in the central aspect of the left broad ligament in the region corresponding to the HSG defect. The lesion was explored but not repaired. Bilateral tubal patency was confirmed via indigo carmine chromopertubation. RESULTS: Ovulation induction and intrauterine insemination were undertaken based on findings at surgery and careful discussion of same with the couple. The patient successfully conceived and had a normal term delivery. CONCLUSION: While HSG abnormalities discovered during infertility assessment are not unusual, intrinsic or de novo peritoneal defects have not been described previously. Abnormal radiographic findings may be explained by this lesion in selected cases. PMID- 12592196 TI - [Tolerance and acceptability of Norplant contraceptive devices]. AB - OBJECTIVES: Norplant implants have been used for long-lasting contraception in Senegal since 1986. The purpose of this work was to assess tolerance and acceptability of the method among Senegalese women. MATERIAL AND METHODS: A prospective non-comparative study including 300 women was undertaken in the Obstetrics and Gynecology Clinic at the University Teaching Hospital A Le Dantec of Dakar between December 1986 and July 1991. Women meeting inclusion criteria were seen at 1 month, 3 months, and 6 months after insertion then every 6-month until device removal. RESULTS: The mean age of women was 31.3+/-5.45 years and mean parity was 4.7+/-2.45. Almost half of the women no longer desired pregnancy and 67.6% of the women who had not used contraceptive method the month before admission decided to use Norplant. During follow-up visits, 45.2% had menstrual side effects mainly amenorrhea 23.4% and irregular bleeding 12.5%. Non-menstrual side effects were local reactions at the implant site (19.9%). The other side effects were dizziness, asthenia, insomnia, anemia, and high blood pressure and weight change. Overall, 58.8% of the women kept their implants and had them removed after 5 years of use. The failure rate was 3.3+/-1.25. At the end of five years of use, satisfaction was 84.8% and nearly two-thirds of the women decided to recommend Norplant to a friend or relative or to use it again. CONCLUSION: Norplant is a method of choice among the range of contraceptive methods available in Senegal. Because of its advantages and its ease of use, Norplant could be a widely used method in Senegal. PMID- 12592198 TI - [Endometriosis of the cervix, colposcopic aspects: a case report and review of the literature]. AB - We present a review of endometriosis of the uterine cervix focusing on colposcopic features, prognosis, and therapeutic strategies. Colposcopic images are provided from a case report. PMID- 12592199 TI - [Pelvic lymphocele: report of a case and review of the literature]. AB - Lymphadenectomy for pelvic cancer can lead to complications, particularly lymphocele. We report a case of pelvic lymphocele, which occurred in a patient who underwent surgery for stage IIa cervical carcinoma after preoperative radiotherapy. The intervention consisted in colpohysterectomy, with lymphadenectomy without peritonisation. Five months later she developed dysuria and pelvic pain. Ultrasound and computed tomography showed a pelvic lymphocele complicated by renal insufficiency. Kidney function was re-established after intraperitoneal marsupialisation. One year later the patient was in good clinical condition with no disorder of kidney function. PMID- 12592200 TI - [Spontaneous hematoma of the rectus sheath during pregnancy: a case report]. AB - A case of spontaneous rectus sheath hematoma is reported in a 32-year-old multigravida transferred to our antepartum unit for premature labor and persistent cough related to Candida dubliniensis upper airway infection. In pregnant patients presenting sudden-onset severe abdominal pain and parietal tenderness, there are two main differential diagnoses: abruptio placentae and aseptic necrobiosis of a uterine leiomyoma. The correct diagnosis may be obtained by ultrasonography and sometimes by computed tomography. In the present case an emergency cesarean section was performed at 35 weeks gestation after a 40% decrease in maternal hemoglobin and onset of fetal heart rate anomalies. PMID- 12592201 TI - [Laboratory tests for women taking oral contraceptives (February 1999)]. PMID- 12592202 TI - [Osteoporosis in menopaused women and patients treated with corticosteroids: diagnostic methods and indications (April 2001)]. PMID- 12592203 TI - Soy and breast cancer. More questions than answers. PMID- 12592204 TI - A new twist on food and drug interactions. PMID- 12592205 TI - Dense breast tissue, mammograms and breast cancer risk. PMID- 12592206 TI - Common painkillers and Alzheimer's disease--more conflicting news. PMID- 12592207 TI - Vaginal problems? Treat yourself to the right solution. AB - You have vaginal itching and discharge, a common problem for many women. You assume it's a yeast infection, but don't be too quick to self-diagnose. Before you treat yourself with an over-the-counter (OTC) medication for a yeast infection, educate yourself about hte signs and symptoms. Often, seeing your health care professional will be the correct first step. Although OTC yeast medications generally work against yeast, they don't work in treating other forms of vaginal problems that may have more serious health consequences. PMID- 12592208 TI - TMJ problems. When your jaw joint acts up. PMID- 12592210 TI - Eating well. Take a healthy coffee break. PMID- 12592211 TI - Why does it seem that I'm getting more moles as I get older? PMID- 12592212 TI - I currently wear bifocal glasses, but I'd like to try contacts. What are my options? PMID- 12592213 TI - Mayo Clinic office visit: breast self-exams. PMID- 12592214 TI - Buprenorphine: an alternative to methadone. PMID- 12592215 TI - Aripiprazole (Abilify) for schizophrenia. PMID- 12592216 TI - [New strategies in the treatment of esophageal cancer]. AB - Esophageal cancer is uncommon, but its incidence is rapidly increasing in the Western countries because of the high incidence of the cardia esophageal adenocarcinoma. Notwithstanding the encouraging results achieved with surgical procedures, esophageal carcinoma has a poor prognosis with 5-year survival in 10% of cases without differences between both squamous and adenocarcinoma histologies. Almost 50% of esophageal cancer patients have unresectable disease at presentation; in the past years combined modality treatments, using chemotherapy and/or radiotherapy with or without surgery, have been evaluated to reduce the risk of local and/or distant recurrences. Ongoing regimens with new agents (Taxanes, Vinorelbine, Irinotecan), in association or not with platinum compounds, show good antitumor efficacy and tolerability, even in the metastatic disease. Preoperative strategies with radiation only did not give any advantage compared to surgery alone, instead, controversial results were obtained, with a minimal advantage, using chemotherapy. Combined chemotherapy and radiation, in suitable candidates for resection has shown an improvement of complete pathological responses, in both the histologies, but with superior toxicities when compared to chemotherapy or radiation therapy alone. Postoperative adjuvant therapies as radiation, chemotherapy or both, have not led to a marked improvement in overall survival and should be performed only in clinical trials. The use of chemoradiotherapy showed a clear advantage versus radiotherapy alone and in many cases equivalent to regimens plus surgery even if control studies haven't been performed. Clinical trials with novel biologic agents, in combination to chemotherapy or alone, against cell growth arrest, neoagiongenesis and tumor metastasis invasion process are currently under evaluation. In the coming years new markers as antigen Ki-67 determination, p53 mutation or high levels activity of thymidylate synthase and novel staging techniques as PET could be precious to identify the better treatment for each patient. PMID- 12592217 TI - Genetic alterations in Barrett's esophagus and esophageal adenocarcinoma. AB - Barrett's esophagus and esophageal adenocarcinoma are increasing health problems in the Western world. The rise in incidence of esophageal adenocarcinoma is greater than that for any other malignancy in Caucasian populations. The social impact of the disease is stressed in addition by the very aggressive nature of esophageal adenocarcinomas with 5-year survival rates of less than 25%. Far more people develop the premalignant condition Barrett's metaplasia than high grade dysplasia and invasive carcinoma. This means that fortunately not all patients with Barrett's metaplasia will make the progression to high grade disease. It is hoped that by unravelling the molecular mechanisms involved in the neoplastic transformation in Barrett's esophagus it will become possible to predict disease progression in the individual patient. This would be a major step forward in the curative treatment of this disease. In addition, identification of the crucial molecular pathways involved in esophageal adenocarcinogenesis would facilitate the development of new treatment strategies. The molecular mechanisms underlying the initiation and progression of this disease are largely unknown. In this review the histological sequence of Barrett's metaplasia via dysplasia to adenocarcinoma is introduced; then the general molecular concepts of carcinogenesis are explained. Furthermore, the most important esophageal neoplasia related genes are described including their possible role in the neoplastic process. The frequent genomic aberrations are put in relation to the different histological entities. Finally, as future prospect, a molecular grading of esophageal adenocarcinogenesis is anticipated. PMID- 12592218 TI - Sub-total oesophagectomy with 2 field lymphadenectomy. AB - At present surgical resection provides the only prospect of long-term survival for oesophageal cancer. It is nevertheless still associated with significant risk although this has dramatically decreased over the last 2 decades. During the same time period, however, the cure rate of both squamous and adenocarcinoma of the oesophagus have failed to improve significantly. There is considerable evidence to suggest that early cancer of the oesophagus (primary tumour confined to the oesophageal wall without lymph node metastases) is associated with a much better prognosis than more advanced tumours. These patients have a good chance of cure with radical surgery. Sub-total oesophagectomy with 2 field lymph node dissection minimises the risk of anastomotic and loco-regional recurrence and improves survival. Nevertheless, the majority of patients will still die of their disease and genuine efforts must be made to determine if patients with a short survival time can be identified and spared unnecessarily aggressive attempts at cure and palliation. PMID- 12592219 TI - The role of tumor suppressor genes in esophageal cancer. AB - Esophageal cancer remains one of the 10 most common cancers worldwide. Although patients with early lesions have a reasonable prognosis, most patients present with advanced disease resulting in an overall 5-year survival of 5-10%. Therefore, the current challenges in the management of esophageal cancer are to obtain a better understanding of the underlying molecular biological alterations to provide new treatment options. During the development of esophageal cancer, there is progression from a premalignant epithelium to a neoplasm that frequently demonstrates a heterogeneous mix of genetic alterations. The vast majority of esophageal cancers have inactivation of the p53 and p16 genes at an early stage followed by defects in genes such as APC, Rb and cyclin D1 at later stages of progression. There is also mounting evidence that numerous, specific regions throughout the genome are frequently lost in these cancers. As a result, we will in the next decade, likely see the discovery and characterizations of novel tumor suppressor genes that may be important in the development of esophageal cancer. The accumulating knowledge about the inactivation of the tumor suppressor genes could ultimately provide us with objective diagnostic tools, more accurate markers for prediction of malignant transformation from premalignant epithelium and facilitate the introduction of novel therapeutic options for the management of esophageal cancer. PMID- 12592220 TI - Management of respiratory symptoms associated with gastroesophageal reflux disease. AB - An association between gastroesophageal reflux and respiratory symptoms has been postulated to exist for some time. Several theories support a pathophysiologic relationship. Effective treatment of gastroesophageal reflux-related respiratory symptoms is not standardized. Modification of lifestyle may be beneficial, but the majority of patients should receive proton pump inhibitor therapy to suppress acid secretion. Treatment failures should be assessed for patient non-compliance, failure to make correct diagnosis, failure to recognize co-existing disease, and failure to treat adequately. If acid suppression is inadequate and acid reflux events persist even with additional acid suppression, patients should be considered for an antireflux operation with expected long-term clinical improvement and decreased medication requirements. Because laparoscopic techniques offer the benefit of antireflux surgery with reduced morbidity, this modality should be considered the current treatment of choice. PMID- 12592221 TI - Prognostic factors relating to esophageal cancer. AB - Prognostic factors for esophageal cancer are difficult to assess because of the lack of standardization and small number of cases over the years. With the increase in number of adenocarcinoma as well as larger amounts of data, there has been a surge in the amount of information regarding this disease. The most significant relationship is between lymph nodes and survival. As the treatment options for esophageal cancer improve the staging of the lymph nodes will become more important in terms of number and ratio. The method of surgical removal is trending toward radical excision, but the prognosis is still poor. With the advent of molecular markers and better chemotherapeutic agents, we will be able to tailor our treatment with respect to the individual tumor. PMID- 12592222 TI - Esophagectomy for esophageal cancer. AB - This review considers the esophagectomy techniques in the treatment of esophageal cancer and provides the guidelines for optimizing the patients' chances at cure, minimizing the risk of mediastinal enteric leak (which carries a mortality rate as high as 50%) and minimizing associated pulmonary insufficiency and infection. The 4 most commonly used routes for resection and replacement include a transhiatal, transthoracic (Ivor-Lewis), tri-incisional (right chest then simultaneous abdominal and left neck), and left chest (distal tumors). Each of these techniques will be described as will the use of colon and jejunum for esophageal replacement. The healthy stomach is the preferred conduit for esophageal replacement. The stomach is well vascularized, easily reaches to the neck, and requires only a single anastomosis for re-establishing intestinal continuity. When the stomach is not available (usually because of prior surgery or disease) the choice of conduits include colon and jejunum. With respect to minimally invasive esophagectomy for esophageal cancer, several groups have significant experience with this and report excellent results. This is an evolving technique but holds much promise for improving the quality of life of patients with cancer without compromising their survival. In locally advanced middle third tumors, thoracotomy and dissection under direct vision's desirable and improves the safety of the operation. The same may apply to tumors receiving neoadjuvant therapy. For tumors of the distal esophagus, transhiatal, tri incisional, and Ivor-Lewis resection are probably equally as safe, and appear to result in equivalent long-term survival. An intrathoracic anastomotic leak is disastrous, carrying a mortality rate of up to 50%. Any surgeon who performs esophagectomy with an intrathoracic anastomosis must do so with a low incidence of leakage, certainly under 5%. Consideration should be given to a cervical placement if there are factors increasing the risk such as the use of induction therapy. PMID- 12592223 TI - Endoscopic ultrasound-guided fine needle aspiration in esophageal cancer. AB - Esophageal cancer has a poor prognosis since it is often diagnosed in the symptomatic and incurable state. Accurate staging at initial diagnosis is imperative as it determines prognosis and influences treatment. Computed tomography (CT) scan is sensitive for identifying metastatic disease but is insensitive for detecting the extent of wall involvement or nodal disease. Endoscopic ultrasound (EUS) has emerged as a powerful tool in staging esophageal cancer with an impressive accuracy. Use of endoscopic ultrasound-guided fine needle aspiration as an adjunct further improves accuracy in nodal staging and allows for histologic confirmation. The impact of this invaluable staging modality in the management of esophageal cancer continues to grow. PMID- 12592224 TI - [Barrett's esophagus. Prevalence, risk of adenocarcinoma, role of endoscopic surveillance]. AB - The presence of gastric metaplasia in the distal esophagus is better known as Barrett's Esophagus (BE). It is an acquired condition caused by gastro-esophageal reflux disease and is associated with a high risk of adenocarcinoma development in the distal esophagus and cardia. The definition of BE has changed over the years as only the specialized metaplasia, with the characteristic "goblet cells", has been shown to carry a risk of cancer development. BE is currently defined as the presence of intestinal metaplasia in the distal esophagus. The prevalence of intestinal metaplasia of the distal esophagus in patients undergoing endoscopy with multiple biopsies for dyspeptic symptoms, varies from 9-21% at the level of the cardia and from 1.2-8% at 3 cm above the esophago-gastric junction, with a decreasing caudo-cranial frequency. Among the BE population (intestinal metaplasia 3 or more cm long) there is a prevalence of male sex and white race, with an average age between the 5(th) and 7(th) decade. The risk of BE mucosa advancing to esophageal adenocarcinoma is not well established: incidence rates from 1/52 years-patient to 1/441 years-patient and a calculated risk from 30 to 125 times higher than in the normal population were reported. These discrepancies are probably related to: 1) temporal differences of the studies, 2) retrospective versus prospective type of the studies, 3) length of follow-up, 4) number of individuals surveilled, 5) regional variations. A literature analysis confirmed that the differences are mostly related to the number of patients studied (the larger the population the lower the incidence), are generally inversely proportional to the follow-up length (the shorter the follow-up the higher the incidence) and depend on the type of the studies (the incidence is higher in the retrospective studies than in the prospective one's). Surveillance program: esophageal adenocarcinoma is a lethal tumor with a 20% 5-year survival rate. The guidelines of The American College of Gastroenterology advice a two-year surveillance rate for BE patients without dysplasia. The difficulty with BE surveillance programs-- even if worthwhile on a single patient basis-- is that they are very expensive and at the present none of the endoscopic surveillance prospective studies has shown a positive impact in the survival rate. From our knowledge it doesn't seem wise to abandon a precautionary surveillance strategy, but further studies are needed to better understand the risk population: at the moment our advice is to monitor male patients in good general conditions with a BE segment longer than 3 cm. PMID- 12592225 TI - Role of FDG-PET scan in staging of cancer of the esophagus and gastroesophageal junction. AB - BACKGROUND: FDG-PET scan has generated in recent years an increasing interest in staging and assessment of response to treatment in cancer patients. METHODS: From the available literature and own data it appears that FDG-PET scan significantly improves detection of haematogenous and distant lymphatic metastasis in carcinoma of the oesophagus and gastro-oesophageal junction (GEJ). Especially diagnostic specificity of lymph node involvement is greatly improved with FDG-PET scan. FDG PET scan also allows a highly sensitive diagnosis of recurrent disease through its capacity of whole-body staging. In assessing response of induction (chemo +/- radiotherapy) in locally advanced disease FDG-PET scan appears to be of high value in predicting response. RESULTS: Moreover, there seems to be a strong correlation between FDG-PET scan response and survival. CONCLUSIONS: It can be concluded that despite the cost, FDG-PET scan should have a place in the algorithm of initial staging, staging of recurrent disease and assessment of response to treatment in cancer of the oesophagus and GE junction. PMID- 12592226 TI - Evaluation of the ELOVL4 gene in patients with autosomal recessive retinitis pigmentosa and Leber congenital amaurosis. AB - PURPOSE: To determine whether pathogenic mutations exist in the ELOVL4 gene in patients with inherited retinal degenerations other than Stargardt-like macular dystrophy or other hereditary macular degenerations. METHODS: All six exons comprising the open reading frame of the ELOVL4 gene were evaluated by single strand conformation analysis, direct nucleotide sequencing, or both methods. RESULTS: No pathogenic mutations were found among 84 patients with autosomal recessive retinitis pigmentosa or among 51 patients with Leber congenital amaurosis (congenital retinal blindness). CONCLUSIONS: These data support the conclusion that recessive retinitis pigmentosa and Leber congenital amaurosis are rarely if ever associated with changes in the ELOVL4 gene. PMID- 12592227 TI - Mutation spectrum of FOXC1 and clinical genetic heterogeneity of Axenfeld-Rieger anomaly in India. AB - PURPOSE: Axenfeld-Rieger anomaly (ARA) is a form of anterior segment dysgenesis of the eye, mainly caused by mutations in the FOXC1 gene. We had earlier reported a novel mutation in the wing region of FOXC1 in an autosomal dominant family. The present study was aimed to identify the spectrum of mutations in the FOXC1 gene in a cohort of Indian ARA patients from different ethnic backgrounds, and to understand its role in the disease pathogenesis. METHODS: Two new autosomal dominant families and seven sporadic cases of ARA from different ethnic backgrounds were screened for mutations by direct sequencing of the coding region of the FOXC1 gene. Another autosomal dominant ARA family that was previously reported by us was also included for comparative analysis of clinical genetic parameters. The segregation of the mutations in the autosomal dominant families was analyzed by haplotype and restriction analysis. Genotype-phenotype correlation were also undertaken to study the role of FOXC1 in phenotypic manifestation in the patient cohort. RESULTS: Three of the nine ARA cases harbored mutations in FOXC1, of which two novel nonsense mutations Q2X and Q123X, resulted in haploinsufficiency of the gene product. The missense mutation (M161K) that we previously reported in an autosomal dominant family was also found in another family. Haplotype analysis of these two families suggested multiple founders in the same ethnic group. The mutations resulted in variable expressions of phenotype among the patients as assessed from their prognosis based on visual outcomes. CONCLUSIONS: Significant genetic heterogeneity of FOXC1 was observed in a multi-ethnic population studied in this region of India resulting in variable ARA phenotypes. The different visual outcome seen in the patients suggest a variable expression of FOXC1 and also provide some insight for understanding the gene functions in this population. PMID- 12592228 TI - Development of the cone photoreceptor mosaic in the mouse retina revealed by fluorescent cones in transgenic mice. AB - PURPOSE: Normal function of the retina relies on the orderly stereotyped organization of different neurons and their synaptic connections. How such neural organization is patterned during development remains poorly understood due to the paucity of adequate developmental markers. This study was to examine the spatial organization and development of cone photoreceptors quantitatively in the mouse retina. METHODS: A transgenic approach was used to generate a living cone cell marker by driving GFP expression in mouse cones with the human red/green opsin gene 5' sequences. The spatial organization and development of the cones in the mouse retinas were examined quantitatively with epifluorescence and scanning laser confocal microscopy. Cone specific GFP expression in the developing retinas was verified with peanut agglutinin (PNA) staining. Developmental expression of mouse cone opsin genes was determined with RT-PCR. RESULTS: The fluorescent retinal cells expressing GFP can be visualized as early as on embryonic day E15. Following up morphological differentiation of these cells revealed features that were consistent with the typical morphology of the mouse cones. Double labeling with cone specific PNA showed that these cells were co-labeled starting from postnatal day P1, and that a subpopulation of PNA positive cones expressed the GFP. The fluorescent cell densities had a similar ventral and dorsal distribution from E15 to P2, increased dramatically in the ventral by P6, and in the dorsal from P7. Nearest neighbor distance analysis demonstrated that this subpopulation of cones was organized into a regular mosaic pattern with a regularity index of 4.82 in the central and 3.55 in the peripheral retina. Quantitative pattern assessment of the developing cones revealed that the fluorescent cells appeared to be distributed in a non-random array before birth. The regularity of the cone array began to rise on P7, in parallel with the onset of mouse green opsin gene expression and the development of cone pedicles. The regular pattern of cone mosaic organization was basically formed by P10, coinciding with the timing of the cone pedicle maturation. CONCLUSIONS: The cones in the mouse retina are organized in a regular mosaic pattern. Patterning the cone mosaic appears to follow a two phase developmental process involving regulated opsin gene expression and cone pedicle maturation: an early phase where a non-random array emerges during cone differentiation, and a late phase where the regular mosaic pattern is mature at the time when cone synaptic contacts are being formed. PMID- 12592229 TI - Supplemental progesterone to prevent preterm birth. PMID- 12592231 TI - The laborist: a new focus of practice for the obstetrician. AB - The practice of obstetrics has changed dramatically in the last decade. Evidence exists for a marked increase in professional dissatisfaction, substance abuse, poor personal relationships, and burnout. These conditions are now being seen in younger physicians and in training programs. Physicians have stopped practicing obstetrics at a much younger age and are increasingly quitting training programs. These findings, along with the recent professional liability insurance crisis, leave many communities with a shortage of physicians who practice obstetrics. A potential solution for alleviating some of these conditions is the introduction of a physician whose sole focus of practice is managing the patient in labor. This physician, called the "laborist," may be able to improve patient care and satisfaction because the laborist will have no other distractions during this time. Also, the laborist will remove from the obstetrician the need to be always available to the laboring patient, which potentially may decrease stress, improve physician well-being, increase length of professional practice, and decrease burnout. PMID- 12592230 TI - Raloxifene and endothelial function in healthy postmenopausal women. AB - OBJECTIVE: The purpose of this study was to determine the effects of raloxifene on endothelial function in healthy, postmenopausal women. STUDY DESIGN: This was a randomized, double-blind, placebo-controlled crossover trial. Subjects (n = 19; mean age, 61 years) underwent endothelial function testing at baseline and after treatment with placebo or raloxifene (60 mg per day for 6 weeks). RESULTS: Brachial artery diameter change (flow-mediated dilation) increased 5.0% with placebo and 8.56% with raloxifene (SE = 1.83, P =.03) in response to a hyperemic stimulus; an adjustment of this response for a variation in stimulus intensity resulted in greater discrimination (P =.009). The ratio of area under the curve response to area under the curve reference with the use of laser Doppler measures was 1.18 for placebo and 1.28 for raloxifene (P =.05). Flow-mediated dilation and area under the curve ratio correlated significantly (r = 0.33, P =.04). CONCLUSION: Treatment with raloxifene enhanced endothelial-mediated dilation in brachial arteries and digital vessels in healthy, postmenopausal women. A potential mechanism for a cardioprotective effect of raloxifene is suggested and warrants further study. PMID- 12592232 TI - Raloxifene does not influence flow-mediated endothelium-dependent and endothelium independent vasodilatation of osteopenic postmenopausal women. AB - OBJECTIVE: The study was undertaken to evaluate whether raloxifene influences endothelium-dependent and endothelium-independent vasodilatation in women. STUDY DESIGN: In a double-blind, placebo-controlled randomized study, 30 osteopenic but otherwise healthy postmenopausal women received either placebo (n = 15 with one dropout) or raloxifene (60 mg/d, n = 15). Brachial artery flow-mediated endothelium-dependent and endothelium-independent vasodilatation was evaluated by ultrasound before and after 6 months of treatment, along with lipid and glucose metabolism. Endothelium-dependent dilatation was evaluated after a 4-minute block of artery blood flow through a cuff insufflated at suprasystolic blood pressure. Endothelium-independent vasodilatation was evaluated after sublingual nitroglycerin administration (400 microg). RESULTS: Placebo did not modify any vascular or metabolic parameter. Raloxifene significantly decreased low-density lipoprotein levels (4.60 +/- 0.18 mmol/L vs 3.94 +/- 0.30 mmol/L, P <.015) but did not modify endothelium-dependent (21.6% +/- 4.6% vs 20.6% +/- 3.8%) or endothelium-independent vasodilatation (26.3% +/- 5.1% vs 32.0% +/- 5.5%). CONCLUSION: Prolonged administration of raloxifene does not influence endothelium dependent and endothelium-independent vasodilatation of osteopenic but otherwise healthy postmenopausal women. PMID- 12592233 TI - Vaginal polymorphonuclear leukocytes and bacterial vaginosis as markers for histologic endometritis among women without symptoms of pelvic inflammatory disease. AB - OBJECTIVE: The study was performed to determine whether vaginal polymorphonuclear leukocytes can be used as predictors of histologic endometritis among women at risk for, but without symptoms of, acute pelvic inflammatory disease. STUDY DESIGN: Five hundred thirty-seven women with, or at risk for, pelvic infection underwent pelvic examinations, including endometrial biopsies. These women were assessed for the presence of vaginal polymorphonuclear leukocytes, bacterial vaginosis, sexually transmitted diseases, and histologic endometritis. RESULTS: Vaginal neutrophils were present in 240 (44.7%) and histologic endometritis was present in 77 (14.3%) of the study population. Women with histologic endometritis were significantly more likely to have vaginal neutrophils present, with an odds ratio of 3.2 (95% CI 1.9-5.7). When study subjects were stratified by the presence of sexually transmitted diseases and vaginal neutrophils, the prevalence of histologic endometritis was highest in women with both conditions present, with an odds ratio of 7.0 (95% CI 3.5-14.3). When subjects were stratified by the presence of bacterial vaginosis and vaginal neutrophils, the prevalence of histologic endometritis was again highest in women with both conditions present, with an odds ratio of 4.8 (95% CI 1.4-16.3). The presence of vaginal neutrophils diagnosed by saline wet mount had a high sensitivity (90.9%) and negative predictive value (94.5%), but a low specificity (26.3%) and positive predictive value (17.1%) for the diagnosis of upper genital tract infection. CONCLUSION: The presence of vaginal polymorphonuclear leukocytes has a high sensitivity and negative predictive value for the diagnosis of upper genital tract infection. PMID- 12592234 TI - Population-based survival for cervical cancer in Singapore, 1968-1992. AB - OBJECTIVE: The aim of this study was to describe the cervical cancer survival rates in Singapore during the period of 1968 through 1992 using population-based data. STUDY DESIGN: A review of cases with cervical cancer diagnosed between 1968 and 1992 from Singapore Cancer Registry was conducted. Information on vital status at December 31, 1997, was available for 3531 cases. Cumulative observed and relative survival rates were computed by using Hakulinen's method. RESULTS: Survival in cervical cancer in Singapore has improved over the 25-year period. The overall 5-year age-standardized relative survival (ASRS) for all patients increased from 45% in 1968-1972 to 65% in 1988-92. Poorer survival rates were observed in elderly patients and Malays. CONCLUSION: The improvement in survival of cervical cancer is likely to be attributed to the rapid national development of Singapore, resulting in the improvements in health services infrastructure and accessibility, as well as to the impact of widely available cytology-based screening resulting in early detection. PMID- 12592235 TI - Metoclopramide pretreatment attenuates emergency contraceptive-associated nausea. AB - OBJECTIVE: The study was undertaken to determine whether metoclopramide pretreatment attenuates side effects associated with high-dose estrogen/progestin emergency contraception in at-risk patients. STUDY DESIGN: This was a randomized, prospective, double-blind, placebo-controlled study at the University of Kansas. Patients (141) requesting emergency contraception and lacking contraindications were offered entry in the study. Both the treatment (metoclopramide 10 mg) and placebo groups received active emergency contraception. Participants evaluated symptom severity with a 12-question survey tool developed for this study. RESULTS: Metoclopramide pretreatment provided protection against nausea and cramping associated with estrogen/progestin emergency contraception. The average scores on a 10-point scale for the treatment group and placebo group were as follows: nausea 3.2 versus 4.8 (P =.01) and cramping 0.9 versus 2.2 (P <.01), indicating less severe nausea and cramping in the treatment group. CONCLUSION: Metoclopramide pretreatment attenuates the nausea and cramping associated with Yuzpe emergency contraceptive treatment. PMID- 12592236 TI - A randomized, double-blind, placebo-controlled, multicenter study that assessed the endometrial effects of norethindrone acetate plus ethinyl estradiol versus ethinyl estradiol alone. AB - OBJECTIVE: The purpose of this study was to determine the incidence of endometrial hyperplasia in subjects who receive continuous norethindrone acetate and ethinyl estradiol combinations versus unopposed ethinyl estradiol. STUDY DESIGN: Nine hundred forty-five postmenopausal women were randomly selected for 12 months of treatment with one of six blinded norethindrone acetate/ethinyl estradiol combinations (milligrams of norethindrone acetate/micrograms of ethinyl estradiol: 0/5, 0.25/5, 1/5, 0/10, 0.5/10, or 1/10) or to open-label 0.625 mg conjugated equine estrogens/2.5 mg medroxyprogesterone acetate. Endometrial hyperplasia and endometrial proliferation were assessed by biopsy at screening, months 6 and 12. RESULTS: Endometrial hyperplasia developed in 26 subjects: Placebo, 0/5 and 0.25/5 (1 subject each) and 0/10 (23 subjects). Significantly less endometrial proliferation was measured in the 1/5 norethindrone acetate/ethinyl estradiol and other norethindrone acetate/ethinyl estradiol combination groups and in the 0.625 mg conjugated equine estrogens/2.5 mg medroxyprogesterone acetate group, than in unopposed ethinyl estradiol groups (6 months: P <.004; 12 months: P <.001). Treatment with 1/5 norethindrone acetate/ethinyl estradiol and with other norethindrone acetate/ethinyl estradiol combinations significantly reduced endometrial proliferation compared with 0.625 mg conjugated equine estrogens/2.5 mg medroxyprogesterone acetate (6 and 12 months: P <.02). CONCLUSION: Norethindrone acetate protects the endometrium from estrogen-induced hyperplasia and changes in proliferative status. In addition, norethindrone acetate/ethinyl estradiol-treated subjects had significantly less endometrial proliferation compared with 0.625 mg conjugated equine estrogens/2.5 mg medroxyprogesterone acetate-treated subjects. PMID- 12592237 TI - Use of health services associated with increased menstrual loss in the United States. AB - OBJECTIVE: The study was conducted to estimate the effect of abnormal uterine bleeding on health care use. STUDY DESIGN: Data were taken from the National Health Interview Survey 1999, a nationwide representative sample of the civilian noninstitutionalized population of the United States. The participants who were eligible for the study were 3133 women aged between 18 and 64 years who reported having a natural menstrual period in the last 12 months and in the last 3 months, never having taken medication containing estrogen (except past use of oral contraceptives), and never been told they had reproductive cancer. From these, 328 women were excluded-5 women because they were more than 64 years old and 323 because of the imprecision of their answers related to their menstrual flow. Analysis was performed with the data from 2805 women: 373 having self-described heavy flow and 2432 having normal flow. RESULTS: Age, family size, insurance coverage, perception of health, and flow of menstrual periods are associated with the majority of questions used to measure health care use (P < or =.05). The odds ratios related to the flow of menstrual periods were all equal to or higher than 1.45 (P < or =.05), which indicates that women who have a heavier flow are at least 1.45 times as likely to use health care as are women who have a lighter or normal flow. CONCLUSION: Women having heavy flow of menstrual bleeding are more likely to use health care than women having a normal flow. PMID- 12592238 TI - Visceral fat is more important than peripheral fat for endometrial thickness and bone mass in healthy postmenopausal women. AB - OBJECTIVE: The purpose of this study was to investigate the influence of body mass index and body composition on endometrial thickness and bone mass. STUDY DESIGN: This was a cross-sectional study that included 531 healthy postmenopausal women aged 48 to 65 years. Endometrial thickness was measured as double-layer thickness. Body composition was measured by dual energy x-ray absorptiometry, which divides the body into fat mass, lean mass, and bone mass, both for the total body and regional body compartments. An abdominal region was inserted manually. Statistics were Pearson correlations and analysis of variance. RESULTS: Endometrial thickness and total body bone mass were correlated, respectively, to body mass index (r = 0.14, P <.01; r = 0.35, P <.001), total body fat mass (r = 0.14, P <.01; r = 0.38, P <.001), abdominal fat mass (r = 0.16, P <.001; r = 0.33, P <.001), peripheral fat mass (r = 0.10, P <.05; r = 0.41, P <.001), and abdominal/peripheral fat mass (r = 0.12, P <.01; r = 0.11, P <.01). CONCLUSION: High body mass index and abdominal fat distribution correlate with increased endometrial thickness and bone mass. PMID- 12592239 TI - Accuracy of detection of trichomonas vaginalis organisms on a liquid-based papanicolaou smear. AB - OBJECTIVE: The study was undertaken to determine the accuracy of the liquid-based Papanicolaou (Pap) smear in the detection of Trichomonas vaginalis organisms in women. STUDY DESIGN: Two hundred three consecutive patients who had the liquid based Pap smear also underwent a culture for T vaginalis. A wet mount was performed if symptomatic. Results were analyzed for sensitivity, specificity, and positive and negative predictive values. RESULTS: Forty-four patients (21.6%) had a positive culture. On the liquid-based Pap smear, 28 patients (13.8%) were positive for T vaginalis, showing a sensitivity of 61.4%, a specificity of 99.4%, a positive predictive value of 96.4%, and a negative predictive value of 90.8%. When wet mount and culture were compared, a sensitivity of 50%, a specificity of 93%, a positive predictive value of 77%, and a negative predictive value of 80% were obtained. CONCLUSION: The presence of T vaginalis organisms, as stated by the liquid-based Pap smear pathology report, is accurate and warrants treatment without further testing. PMID- 12592240 TI - Ethnic differences at risk for gestational trophoblastic disease in New Mexico: A 25-year population-based study. AB - OBJECTIVE: The purpose of this study was to compare gestational trophoblastic disease incidence rates with the use of population-based data. STUDY DESIGN: All incident cases between 1973 and 1997 and live birth, pregnancy, and women at risk were tabulated with the use of data that were derived from the New Mexico Tumor Registry and Vital Records and Health Statistics Annual Reports. Statistical methods included trends analyses, odds ratios, and Poisson regression. RESULTS: Of 939 total cases, 312 non-Hispanic white women, 399 Hispanic white women, 201 American Indian women, and 27 other women were affected. Age-adjusted incidence rates were significantly higher for American Indian women (11.16%) compared with non-Hispanic (3.57%) or Hispanic white women (5.32%); the probability value was <.001. When live birth (1:438 women) and pregnancy (1:486 women) denominators were considered, American Indian women alone were at increased risk, and the ratio increased by 56% over 25 years. American Indian women were also at increased risk for partial mole (relative risk, 4.03; 95% CI, 2.57-6.31), invasive mole (relative risk, 26.7; 95% CI, 7.81-93.14), and choriocarcinoma (relative risk, 6.29; 95% CI, 1.81-22.66) variants. CONCLUSION: American Indians are at increased risk relative to the other predominant ethnic groups in New Mexico. Age-adjusted standardization provided a reproducible measurement that may be applicable across other registries. PMID- 12592241 TI - Laparoscopic preservation of ovarian function: an underused procedure. AB - OBJECTIVE: There are many young women undergoing irradiation or chemotherapy without having the option of preserving their ovarian function. Our purpose was to review the literature on laparoscopic ovarian transposition, to evaluate its efficacy, and to provide clinical opinion on the subject. STUDY DESIGN: We evaluated the English articles on laparoscopic ovarian transposition identified through a MEDLINE search. We also report a case of laparoscopic ovarian tranposition in a 28-year-old woman with rectal cancer. RESULTS: Laparoscopic ovarian transposition in women <40 years old is associated with preservation of ovarian function in 88.6% of cases. During and afer irradication, our patient never missed her menstrual periods. CONCLUSION: Despite its simplicity and efficacy, this procedure is underused. We encourage clinicians to discuss and propose it to their patients. PMID- 12592242 TI - The outcome of in vitro fertilization in advanced endometriosis with previous surgery: a case-controlled study. AB - OBJECTIVE: This study was undertaken to evaluate the outcome of in vitro fertilization (IVF) in patients with advanced pelvic endometriosis and previous surgical treatment. STUDY DESIGN: A case controlled study was performed. RESULTS: Patients with the diagnosis of stage IV endometriosis with previous surgical treatment were treated by IVF (group A = 85). An age-matched group of patients (group B = 177) with tubal factor infertility were treated with the same protocol of IVF. In group A, cycle cancellation because of poor response occurred in 29.7% compared with 1.1% in the control group (relative risk 26.03, 95% CI 6.02 112.45). There were 13 (15.3%) clinical pregnancies per stimulated cycle in group A compared with 93 (52.5%) clinical pregnancies in the control group, P <.0001 (odds ratio 0.29, 95% CI 0.15-0.55). CONCLUSION: The outcome of IVF in stage IV endometriosis with previous surgery was significantly lower compared with an age matched group of tubal factor infertility. PMID- 12592243 TI - Recruitment of CD4 T lymphocytes and macrophages into the cervical epithelium of women after coitus. AB - OBJECTIVE: T lymphocytes and macrophages are considered essential components of the immune response. Many factors are known to influence the presence and distribution these cells in genital mucosa. This study investigated the effect of sexual intercourse on cervical intraepithelial T lymphocytes and macrophages in healthy uninfected women. STUDY DESIGN: Cervical intraepithelial samples were obtained with an endocervical brush from 31 women; the cervical T lymphocytes and macrophages were analyzed by flow cytometry. Eleven women with a history of last sexual intercourse at <3 days were compared against 20 women with last sexual intercourse of >3 days. Furthermore, cellular activation markers (CD69, CD25, HLA DR) on T lymphocytes and costimulatory molecules (CD80, CD86) on macrophages were studied. RESULTS: Women with last sexual intercourse at <3 days showed predominance of CD4(+) T lymphocytes compared with women with last sexual intercourse of >3 days (P <.02); the numbers of macrophages were higher in the latter (P <.005). No difference was found in the density of T-lymphocyte activation and macrophage costimulatory markers between the two cohorts. CONCLUSION: Within cervical epithelium, the distribution of mononuclear leucocytes may be altered after coitus. The higher proportion of cervical intraepithelial CD4(+) T cells that were observed in the early postcoital period suggests a mechanism by which the relative risk of the acquisition of human immunodeficiency virus infection is increased in women. PMID- 12592244 TI - Are women with urogenital atrophy symptomatic? AB - OBJECTIVE: The purpose of this study was to determine the degree of correlation between physical signs of genital atrophy and symptoms that are suggestive of atrophic vaginitis. STUDY DESIGN: Female volunteers (n = 135; mean age, 69 years) rated the presence and severity (rating, 0-3) of vaginal atrophy symptoms. The presence and severity of vaginal mucosal changes, which included vaginal pH (0 3), were recorded during a pelvic examination. A vaginal cytologic maturation value was performed. Symptoms, signs, pH, and maturation value were correlated by the Spearman rank test. RESULTS: Symptom scores were low (mean, 0.41; range, 0 2.6). Symptoms were only weakly correlated with physical findings (r = 0.14) and not with maturation value (r = 0.06) or age (r = -0.004). There was a moderate correlation between physical examination score and maturation value (r = -0.48). In women > or =65 years old, symptom score and physical examination score were correlated weakly (r = 0.25). Low pH correlated well with high maturation value (r = -0.52). Women who were undergoing estrogen therapy had higher symptoms scores (P =.0007) and maturation values (P =.0002) than women who were not undergoing therapy. CONCLUSION: Although urogenital atrophy occurs universally after menopause, most elderly women are minimally symptomatic. Those women on estrogen replacement therapy may be more symptomatic. Symptoms alone should not be used as a guide for the initiation of estrogen therapy. PMID- 12592245 TI - Comparison of the effect of raloxifene and continuous-combined hormone therapy on mammographic breast density and breast tenderness in postmenopausal women. AB - OBJECTIVE: The objectives were to evaluate mammographic changes in breast density that are associated with raloxifene or hormone therapy and to compare the safety profiles of the two therapies. STUDY DESIGN: Postmenopausal women older than 60 years who had a bone mineral density T-score of < or =-1 were assigned randomly to receive raloxifene hydrochloride 60 mg/day or continuous-combined hormone therapy that consisted of conjugated equine estrogen 0.625 mg/day plus medroxyprogesterone acetate 2.5 mg/day in a 1-year, open-label study. Radiologists who were blinded to the treatment assignment assessed the mammograms according to the American College of Radiology Breast Image Reporting and Data System breast density categories. Breast density changes were analyzed for treatment differences. RESULTS: After 12 months of treatment, 0.9% of the women who received raloxifene had increased mammographic breast density compared with 27.4% of the women who received continuous-combined hormone therapy (P <.001). In the continuous-combined hormone therapy group, 77% of the women reported breast tenderness at any time during the study, compared to 22% of the women in the raloxifene group. CONCLUSION: In postmenopausal women with low bone mass, raloxifene therapy for 12 months does not increase mammographic breast density, whereas continuous-combined hormone therapy substantially increases breast density in a significant number of women. PMID- 12592246 TI - A comparison of four screening methods for cervical neoplasia in a developing country. AB - OBJECTIVE: The purpose of this study was to compare the results of cytologic examination, cervicography, direct acetic acid test, and magnified acetic acid test (speculoscopy) with the results of histologic examination. STUDY DESIGN: The study was a prospective, descriptive, and analytic study that included 1286 women in a primary health facility who were screened by cytologic examination, cervicography, direct acetic acid test, and magnified acetic acid test (speculoscopy). Histologic evidence was obtained on all patients by means of cervical punch biopsies. The biopsy specimens were taken from aceto-white areas or areas suspicious of cancer. In normal individuals, the specimens were taken from the external cervical os at the 12 or 6 o'clock position, whichever was most convenient. Positive cases were treated by colposcopy and large loop excision of the transformation zone. Testing for human immunodeficiency virus infection was not done. RESULTS: Of the 1286 women who were screened, 1093 women (85%) were eligible for analysis. The mean age of the women was 38.6 years. Because histologic evidence was obtained on all women, direct sensitivities and specificities could be calculated for each screening method. The sensitivity, specificity, and positive predictive values for cytologic examination were 53%, 95%, and 47%, respectively; for cervicography were 49%, 88%, and 26%, respectively; for the direct acetic acid test were 79%, 49%, and 12%, respectively; and for speculoscopy were 77%, 45%, and 11%, respectively. Of the 876 patients who were eligible for colposcopy, only 468 women (56%) eventually returned for colposcopy. CONCLUSION: Cytologic examination, cervicography, the direct acetic acid test, and speculoscopy are not suited for screening in developing countries because of the low sensitivity of the cytologic examination and the low specificity of the other tests. Two or more tests combined will increase the sensitivity, but the specificity remains low. Screening methods in developing countries remain an urgent problem. PMID- 12592247 TI - Can ultrasound replace dilation and curettage? A longitudinal evaluation of postmenopausal bleeding and transvaginal sonographic measurement of the endometrium as predictors of endometrial cancer. AB - OBJECTIVE: The purpose of this study was to evaluate postmenopausal bleeding and transvaginal sonographic measurement of endometrial thickness as predictors of endometrial cancer and atypical hyperplasia in women whose cases were followed for > or =10 years after referral for postmenopausal bleeding. STUDY DESIGN: Women (n = 394) who had postmenopausal bleeding from November 1987 to October 1990 underwent transvaginal sonographic measurement of endometrial thickness and curettage. It was possible to assess the medical records (regarding recurrence of a postmenopausal bleeding, development of endometrial cancer, and death) in 339 of the 394 women (86%) > or =10 years after referral for postmenopausal bleeding. RESULTS: Thirty-nine of the 339 women (11.5%) had endometrial cancer, and 5 women (1.5%) had atypical hyperplasia. The relative risk of endometrial cancer in women who were referred for postmenopausal bleeding was 63.9 (95% CI, 46.0-88.8); the corresponding relative risk for endometrial cancer and atypical hyperplasia together was 72.1 (95% CI, 52.8-98.5) compared with women of the same age from the general population of the same region of Sweden. No woman with an endometrial thickness of < or =4 mm was diagnosed as having endometrial cancer. The relative risk of the development of endometrial cancer in women with an endometrial thickness of >4 mm was 44.5 (95% CI, 6.5-320.1) compared with women with an endometrial thickness of < or =4 mm. The reliability of endometrial thickness (cutoff value, < or =4 mm) as a diagnostic test for endometrial cancer was assessed: Sensitivity, 100%; specificity, 60%; positive predictive value, 25%; and negative predictive value, 100%. The incidence of endometrial cancer or atypical hyperplasia in women with an intact uterus whose cases had been followed for > or =10 years was 5.8% (15/257 women) compared with 22.7% (15/66 women) in women who had < or =1 episode of recurrent bleeding. No endometrial cancer was diagnosed in women with a recurrent postmenopausal bleeding who had an endometrial thickness of < or =4 mm at the initial scan. CONCLUSION: Postmenopausal bleeding incurs a 64-fold increase risk for endometrial cancer. There was no increased risk of endometrial cancer or atypia in women who did not have recurrent bleeding, whereas women with recurrent bleeding were a high-risk group. No endometrial cancer was missed when endometrial thickness measurement (cutoff value, < or =4 mm) was used, even if the women were followed up for < or =10 years. We conclude that transvaginal sonographic scanning is an excellent tool for the determination of whether further investigation with curettage or some form of endometrial biopsy is necessary PMID- 12592248 TI - Nucleated red blood cells in infants of mothers with asthma. AB - OBJECTIVE: The purpose of this study was to evaluate whether the absolute nucleated red blood cell and lymphocyte count is elevated in term, appropriate for-gestational-age infants born to women with asthma. STUDY DESIGN: We compared absolute nucleated red blood cell counts taken during the first 12 hours of life in two groups of term, vaginally delivered, appropriate-for-gestational-age infants; one group was born to mothers with active asthma during pregnancy (n = 28 infants), and the other group was born to control mothers (n = 29 infants). Asthma severity was classified according to the National Asthma Education and Prevention Program. We excluded infants of women with diabetes mellitus, hypertension, alcohol, and tobacco or drug abuse and infants with fetal heart rate abnormalities, hemolysis, blood loss, or chromosomal anomalies. RESULTS: There were no differences between groups in birth weight, gestational age, maternal age, gravidity, parity, maternal analgesia during labor, 1- and 5-minute Apgar scores, and infant sex. The hematocrit level, red blood cell count, absolute nucleated red blood cell count, and corrected leukocyte and lymphocyte counts were significantly higher in the asthma group than in the control group. The platelet count was not significantly different between groups. The absolute nucleated red blood cell count correlated significantly with the asthma severity score (r (2) = 28%, P <.001). Backward stepwise multiple regression that included Apgar scores and gestational age showed a significant correlation of absolute nucleated red blood cell count with the presence of asthma and its severity (P <.001). CONCLUSION: At birth, term appropriate-for-gestational-age infants born to mothers with asthma have increased circulating absolute nucleated red blood cell and lymphocyte counts compared with control infants. PMID- 12592249 TI - Hemodynamic changes during complete umbilical cord occlusion in fetal sheep related to hippocampal neuronal damage. AB - OBJECTIVES: The purpose of our study was to examine the physiologic changes caused by 10 minutes of umbilical cord occlusion in fetal sheep and to determine the correlation between fetal acidemia or cerebral ischemia and hippocampal neuronal damage. STUDY DESIGN: Thirteen fetal sheep were instrumented and catheterized. Carotid artery blood flow (CaF), fetal mean arterial blood pressure (FMABP), pH, PCO (2), base excess, oxygen saturation (SatO(2)), and PO (2) were monitored throughout the occlusion study. Brain sections were examined for the hippocampal neuronal damage. RESULTS: Our data showed severe ischemia (CaF: 10 +/ 7 mL/min; FMABP: 29 +/- 8 mm Hg) and acidemia (pH: 7.0 +/- 0.05; base excess: 9.9 +/- 2.4 mEq/L) at the end of occlusion. The neuronal damage score had significant correlations with ischemia and also with reperfusion, but not with the acidemic or hypoxic parameters. CONCLUSION: We demonstrated that the degree of hippocampal damage was correlated with the degree of ischemia and reperfusion. PMID- 12592250 TI - Prophylactic administration of progesterone by vaginal suppository to reduce the incidence of spontaneous preterm birth in women at increased risk: a randomized placebo-controlled double-blind study. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of prophylactic vaginal progesterone in decreasing preterm birth rate in a high-risk population. STUDY DESIGN: A randomized, double-blind, placebo-controlled study included 142 high-risk singleton pregnancies. Progesterone (100 mg) or placebo was administered daily by vaginal suppository and all patients underwent uterine contraction monitoring with an external tocodynamometer once a week for 60 minutes, between 24 and 34 weeks of gestation. Progesterone (n = 72) and placebo (n = 70) groups were compared for epidemiologic characteristics, uterine contraction frequency, and incidence of preterm birth. Data were compared by chi(2) analysis and Fisher exact test. RESULTS: The preterm birth rate was 21.1% (30/142). Differences in uterine activity were found between the progesterone and placebo groups (23.6% vs 54.3%, respectively; P <.05) and in preterm birth between progesterone and placebo (13.8% vs 28.5%, respectively; P <.05). More women were delivered before 34 weeks in the placebo group (18.5%) than in the progesterone group (2.7%) (P <.05). CONCLUSION: Prophylactic vaginal progesterone reduced the frequency of uterine contractions and the rate of preterm delivery in women at high risk for prematurity. PMID- 12592251 TI - Parental occupations and other risk factors associated with nonchromosomal single, chromosomal single, and multiple birth defects: a population-based study in Singapore from 1994 to 1998. AB - OBJECTIVE: The purpose of this study was to evaluate the association between the prevalence of birth defects among maternal and paternal occupation groups in Singapore for live births between January 1, 1994, and December 31, 1998, and to determine whether there are certain demographic risk factors and maternal and/or paternal occupation groups that are associated with an increased risk for birth defects versus chromosomal single birth defect, nonchromosomal single birth defect, and multiple birth defects. STUDY DESIGN: This was a retrospective study. Information on live births (Singapore National Registry of Births and Deaths) and birth defect cases (National Birth Defects Register) were studied. Other information included the mother's date of birth, ethnic group, highest educational qualification, and the occupation of the mother and the father. Cox regression analysis was used to determine whether certain demographic and occupational factors were associated with the three groups of birth defects. RESULTS: Of a total of 237,755 live births, there were 3276 birth defect cases (nonchromosomal single birth defect, 1869 cases; chromosomal single birth defect, 197 cases; and multiple birth defects, 1210 cases). Increased risks for all types of birth defects were observed with advancing gestational age. Occurrence of nonchromosomal single birth defect and multiple birth defects were significantly higher for multiple births compared to singleton births. Significant associations were found with the use of "legislators, senior officers, and managers" as reference: Maternal occupation of "cleaners, laborers, and related workers" with chromosomal single birth defect anomalies (adjusted risk ratio, 4.86; 95% CI, 1.07-22.14); paternal occupation of "plant and machine operators and assemblers" (adjusted risk ratio, 1.50; 95% CI, 1.14-1.98) with nonchromosomal single birth defect; "production craftsmen and related workers" and "cleaners and laborers and related workers" with nonchromosomal single birth defect (adjusted risk ratio, 1.42; 95% CI, 1.10-1.82; and adjusted risk ratio, 1.43; 95% CI, 1.07-1.91 respectively); and multiple birth defects (adjusted risk ratio, 1.42; 95% CI, 1.03-1.94; and adjusted risk ratio, 1.47; 95% CI, 1.03-2.09, respectively). CONCLUSION: Maternal delivery age is an important risk factor for all birth defects. Mothers and fathers who work as "cleaners and laborers and related workers" appear to have a higher risk of giving birth to children with chromosomal single birth defect and nonchromosomal single birth defect and multiple birth defects, respectively. Further in-depth study would be needed to confirm these observations. PMID- 12592252 TI - Basal metabolic rate in hyperemesis gravidarum: comparison to normal pregnancy and response to treatment. AB - OBJECTIVE: The aim of the study was to measure resting minute ventilation and oxygen consumption in patients with hyperemesis gravidarum before and after treatment and to compare the results with those of normal pregnant women. STUDY DESIGN: Baseline evaluation was performed with the use of an open-circuit ventilatory system in 17 hospitalized patients with hyperemesis gravidarum and was repeated 1 week after treatment. Thirty-seven normal pregnant women served as control subjects. RESULTS: Resting minute ventilation and oxygen consumption were decreased by 20% and 15% (P <.001), respectively, in patients with hyperemesis compared with control subjects. The decrease occurred despite a measurable increase of free thyroxine compared with normal pregnancy. Treatment resulted in a 10% increase in oxygen consumption in those patients with increased thyroid stimulation, but not otherwise. CONCLUSION: Resting oxygen consumption is reduced significantly in patients with hyperemesis gravidarum that is consistent with a state of partial starvation rather than increased in parallel with increases of thyroid axis activity. PMID- 12592253 TI - Systemic administration of betamethasone delays endotoxin-induced preterm labor in the murine model. AB - OBJECTIVE: The purpose of this study was to determine whether the administration of betamethasone decreases the endotoxin-induced preterm parturition rate and inhibits the risk of cytokines in the murine model. STUDY DESIGN: Endotoxin was administered intraperitoneally at gestational day 15 (75% of gestation). In phase I, the duration of gestation was measured in 36 gravid C3H/HeOu mice that were equally divided into four treatment groups: control, endotoxin only, and two different dose regimens of betamethasone followed by endotoxin. In phase II, maternal serum and amniotic fluid concentrations of cytokines (interleukin 1alpha, tumor necrosis factor-alpha, and interleukin-6) were measured at 4 hours after endotoxin injection in 44 gravid mice divided equally in the four treatment groups. RESULTS: The group that was exposed only to endotoxin was delivered at a significantly earlier gestational age compared with the control group (16.2 +/- 0.4 days vs 19.6 +/- 0.2 days; P <.01). The two groups that were pretreated with betamethasone before the endotoxin were delivered at gestational ages similar to the control group. There was a marked increase of tumor necrosis factor-alpha and interleukin-6 levels in amniotic fluid of mice that were treated with endotoxin only compared with the control group (P <.001). No difference in cytokine levels was found in those mice that were premedicated with betamethasone compared with the control group. CONCLUSION: Antenatal administration of betamethasone to mice delayed preterm parturition that was induced by endotoxin. Elevations of amniotic fluid cytokine concentrations that were observed with endotoxin were not observed with pretreatment with betamethasone. PMID- 12592255 TI - Longitudinal evaluation of markers of endothelial cell dysfunction and hemostasis in treated antiphospholipid syndrome and in healthy pregnancy. AB - OBJECTIVE: The purpose of this study was to determine whether primary antiphospholipid syndrome pregnancies are associated with endothelial cell activation in the maternal circulation. STUDY DESIGN: Markers of endothelial cell activation were measured every 4 weeks during pregnancy in the blood of 23 women with primary antiphospholipid syndrome and 19 control subjects. All women with antiphospholipid syndrome received anticoagulant treatment. Plasma concentrations of plasminogen activator inhibitor type 1, tissue plasminogen activator, soluble intercellular adhesion molecule-1, vascular cell adhesion molecule-1, E-selectin, and soluble thrombomodulin were determined by enzyme-linked immunoassay. Concentrations of prothrombin fragments 1+2 and D-dimers were also determined. RESULTS: Three antiphospholipid syndrome pregnancies (13%) were complicated by intrauterine growth restriction and preeclampsia; one antiphospholipid syndrome pregnancy (4%) was complicated by preterm rupture of membranes. Six women with antiphospholipid syndrome (26%) had thrombotic events. Differences in concentrations of endothelial cell activation markers between antiphospholipid syndrome and control pregnancies were not significant. CONCLUSION: Despite poorer pregnancy outcome, there was no evidence of greater endothelial cell activation in antiphospholipid syndrome pregnancies that were treated. PMID- 12592254 TI - Single and multiple prenatal glucocorticoid exposures improve preterm newborn lamb cardiovascular and renal function similarly. AB - OBJECTIVE: Renal and cardiovascular function improves in preterm newborn lambs after a single prenatal betamethasone treatment. We hypothesized that multiple betamethasone exposures would further improve renal and cardiovascular adaptation. STUDY DESIGN: Pregnant ewes were chosen randomly to receive saline solution, one dose of 0.5 mg/kg betamethasone at 104 days of gestation, or three doses of 0.5 mg/kg betamethasone at 104, 111, and 118 days of gestation. Lambs were delivered at 125 days of gestation (preterm) or 145 days of gestation (term). Renal and cardiovascular responses to phenylephrine were evaluated at 2 hours of age. RESULTS: The preterm single and multiple betamethasone-treated lambs comparably increased glomerular filtration rate, urinary flow and osmolar clearance, and sodium excretion in response to phenylephrine. Term responses were similar and not influenced by betamethasone exposure. CONCLUSION: Multiple courses of betamethasone do not further improve renal and cardiovascular responses from a single betamethasone dose. Renal and cardiovascular function at term is not affected by early prenatal betamethasone exposure. PMID- 12592256 TI - Antepartum fetal asphyxia in the preterm pregnancy. AB - OBJECTIVE: The objective of this report was to provide insight into the frequency and characteristics of antepartum fetal asphyxia in pregnancies that are delivered preterm. STUDY DESIGN: The characteristics of 30 pregnancies that were delivered preterm with biochemically confirmed antepartum fetal asphyxia (umbilical artery base deficit of >12 mmol/L) that were derived from >1 decade of experience in a single tertiary care obstetric unit were examined. Antepartum clinical characteristics, fetal assessment tests, and neonatal complications were documented. Fetal asphyxia was classified as mild, moderate, or severe on the basis of an umbilical artery base deficit (>12 mmol/L) and newborn encephalopathy and other organ system complications. RESULTS: Antepartum fetal asphyxia accounted for at least 34% of the fetal asphyxia in the pregnancies that were delivered preterm. Predictive criteria that led to intervention and diagnosis included clinical risk factors and, particularly, abnormal fetal assessment tests. The 50% incidence of moderate or severe asphyxia in the antepartum preterm pregnancies compares with 15% in term pregnancies. Moderate or severe asphyxia occurred with equal frequency with early and delayed intervention. CONCLUSION: Fetal asphyxia in pregnancies that were delivered preterm is present frequently before the onset of labor. Abnormal fetal assessment tests are valuable predictors of antepartum fetal asphyxia. The increased frequency of moderate and severe fetal asphyxia in the pregnancy that is delivered preterm implies a greater likelihood of long-term morbidity or death. PMID- 12592257 TI - Effects of steroid hormones on calcitonin gene-related peptide receptors in cultured human myometrium. AB - OBJECTIVE: This study was designed to examine whether, with the use of human myometrial explants in culture, calcitonin gene-related peptide B receptors are regulated by steroid hormones. STUDY DESIGN: Myometrial tissues were obtained from the lower uterine segments from women in a nonpregnant state and pregnant women who were not in labor undergoing cesarean delivery (39.1 +/- 0.2 weeks of gestation). Tissues were incubated in Dulbecco's modified Eagle's medium without phenol red, with either 17beta-estradiol (10(-9), 10(-8), 10(-7) mol/L), progesterone (10(-8), 10(-7), 10(-6) mol/L), or in a combination of 17beta estradiol (10(-8) mol/L) and progesterone (10(-7) mol/L). Reverse transcriptase polymerase chain reaction was performed to examine the expression of estrogen receptor-alpha, progesterone receptor, and Western blotting was used for calcitonin gene-related peptide B receptor protein measurement in the myometrium. RESULTS: We found that (1) messenger RNA expression for both estrogen receptor alpha and progesterone receptor in nonpregnant myometrium significantly declined within 48 hours explant culture; (2) estrogen receptor-alpha messenger RNA levels in the pregnant myometrium were 98.6%, 95.3%, and 89.8% at 12, 24, and 48 hours of incubation, respectively; (3) the levels of messenger RNA for progesterone receptor in the pregnant myometrium were 96.2%, 93.3%, 90.1%, at 12, 24, and 48 hours of incubation, respectively; (4) the 17beta-estradiol dose dependently inhibited calcitonin gene-related peptide B receptor protein in pregnant myometrium explant culture; (5) the progesterone dose dependently increased calcitonin gene-related peptide B receptor expression in pregnant myometrium explant culture; (6) the combined treatment with 17beta-estradiol (10(-8) mol/L) and progesterone (10(-7) mol/L) further enhanced myometrial calcitonin gene related peptide B receptor protein expression. CONCLUSION: We conclude from this study that (1) the messenger RNA expression of estrogen receptor-alpha and progesterone receptor in pregnant myometrium were maintained at relatively high levels (>89%) within 48 hours incubation and that this may be useful for in vitro studies that are designed to evaluate the effects of sex steroids on the human myometrium during pregnancy and that (2) estrogen inhibits and progesterone stimulates the expression of calcitonin gene-related peptide B receptors in cultured pregnant myometrial explants. We suggest that steroid hormone-regulated calcitonin gene-related peptide B receptor expression could underlie differential myometrial sensitivity to calcitonin gene-related peptide-induced relaxation. PMID- 12592258 TI - Soluble HLA-DR levels in the maternal circulation of normal and pathologic pregnancy. AB - OBJECTIVE: The purpose of this study was to determine that circulating HLA-DR molecules are important candidates for the monitoring of maternal immunostimulation and immunosuppression. STUDY DESIGN: Concentrations of soluble HLA-DR molecules were estimated in EDTA plasma samples of 61 nonpregnant women, 123 healthy pregnant women in the second trimester, 66 healthy women who were delivered at term, and 136 women who were delivered because of complications such as uncontrollable preterm intrauterine activation, abruptio placentae, intrauterine growth retardation, preeclampsia, and HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome. RESULTS: In comparison to nonpregnant women, the normal course of pregnancy was associated with strongly increasing levels of soluble HLA-DR from second trimester on until term. In comparison to women who were delivered preterm because of uncontrollable intrauterine activation, increased soluble HLA-DR concentrations were detected in case of HELLP syndrome (P <.05), although decreased levels were detected in the case of intrauterine growth retardation, preeclampsia (P <.01), and abruptio placentae (P <.01). CONCLUSION: Dysregulation of the maternal immune response to pregnancy may play an important role in the cause of complicated pregnancies. PMID- 12592259 TI - Cell-free fetal DNA in the plasma of pregnant women with severe fetal growth restriction. AB - OBJECTIVE: Although there have been reports of increased fetal nucleated erythrocytes in the blood of pregnant women who are carrying growth-restricted fetuses, there have been no reports of quantification of fetal DNA concentration in the plasma of women with fetal growth restriction. We quantified fetal DNA concentration in the plasma of pregnant women with preeclampsia and/or fetal growth restriction. STUDY DESIGN: We examined maternal plasma from 9 pregnant women with fetal growth restriction and 9 with preeclampsia and from 20 women who were gestational age-matched normal control subjects. All women carried a male fetus. DNA was extracted from 1.5-mL plasma samples, and the DYS14 and beta globin gene were analyzed by real-time quantitative polymerase chain reaction. RESULTS: The concentration of fetal DNA was significantly higher in subjects with preeclampsia than in fetal growth restriction subjects and normal control subjects. Fetal DNA concentrations in fetal growth restriction subjects were similar to those of normal control subjects. The concentration of total DNA (beta globin) was significantly higher in subjects with preeclampsia when compared with healthy control subjects. CONCLUSION: We demonstrated that there was no increase in fetal DNA in the plasma of pregnant women with fetal growth restriction and that most fetal DNA in maternal plasma originates from trophoblasts. PMID- 12592260 TI - Regulation of insulin-like growth factor-I and insulin-like growth factor binding protein-1 concentrations in preterm fetuses. AB - OBJECTIVE: Our purpose was to evaluate which factors regulate insulin-like growth factor-I and insulin-like growth factor binding protein-1 concentrations in preterm fetuses. STUDY DESIGN: We studied 76 singleton births between 25 and 36 weeks of gestation. Forty-nine pregnancies were complicated by hypertensive disease; 24 pregnancies were complicated by preterm labor or preterm rupture of membranes; and antenatal glucocorticoids were given in 49 pregnancies. Pathology reports showed infarct(s) or hematoma(s) in 31 of 69 placentas. We recorded blood gas values in umbilical artery and vein and measured glucose, C-peptide, and insulin-like growth factor-I and insulin-like growth factor binding protein-1 concentrations in umbilical vein. RESULTS: Birth weight correlated with umbilical vein insulin-like growth factor-I (r = 0.68, P <.0001) and inversely with insulin like growth factor binding protein-1 (r = -0.26, P =.02). Babies with birth weight of 25th percentile. Two-factor analysis of variance showed that umbilical vein insulin like growth factor-I was determined by gestational age (P =.0004) and birth weight percentile (P <.0001), whereas insulin-like growth factor binding protein 1 was not affected by gestational age. Umbilical vein C-peptide was highly correlated with insulin-like growth factor binding protein-1 (r = -0.55, P <.0001), but not insulin-like growth factor-I, levels. Blood gas values in umbilical artery and vein, particularly umbilical artery PO (2), were correlated with umbilical vein insulin-like growth factor-I and insulin-like growth factor binding protein-1 (r = 0.51 and -0.48, respectively; P <.0001); changes in insulin-like growth factor-I and insulin-like growth factor binding protein-1 occurred at umbilical artery PO (2) <14.8 mm Hg. Multiple regression analysis showed that umbilical vein insulin-like growth factor-I was predicted by umbilical artery PO (2), gestational age, and the presence of placental infarcts/hematomas (R (2) of model = 0.58, P <.0001), and umbilical vein insulin like growth factor binding protein-1 by umbilical vein C-peptide, umbilical artery PO (2), and placental infarcts/hematomas (R (2) = 0.49, P <.0001). CONCLUSION: In the preterm fetus, circulating insulin-like growth factor-I is related to gestational age and the in utero growth potential, whereas insulin like growth factor binding protein-1 is related only to the in utero growth potential. The PO (2) is a robust determinant of both insulin-like growth factor I and insulin-like growth factor binding protein-1 levels; hypoxia may restrain fetal growth through its effects on the insulin-like growth factor/insulin-like growth factor binding protein axis. Insulin is a powerful determinant of insulin like growth factor binding protein-1, but not insulin-like growth factor-I, concentrations in the preterm fetus. PMID- 12592261 TI - Second-opinion magnetic resonance imaging for suspected fetal central nervous system abnormalities. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship of magnetic resonance imaging and gestational age in the setting of fetuses with suspected abnormalities of the central nervous system that were detected by ultrasound scanning. STUDY DESIGN: Multiplanar magnetic resonance studies were performed in fetuses with suspected central nervous system abnormalities on ultrasound scanning. Magnetic resonance imaging was evaluated for its ability to provide additional information, change the diagnosis, or impact obstetric treatment. Patients were grouped by gestational age at the time of magnetic resonance imaging. RESULTS: Magnetic resonance imaging provided additional information in 46 of 72 pregnancies (64%), changed the diagnosis in 20 of 72 pregnancies (28%), and potentially altered the timing or mode of delivery in 8 of 72 pregnancies (11%). Additional information increased with increasing gestational age groups (P =.03). CONCLUSION: Magnetic resonance imaging provided additional information in two thirds of the fetuses with central nervous system abnormalities, which was significantly increased with increasing gestation. Antenatal treatment was influenced by magnetic resonance imaging in 11% of the cases. PMID- 12592262 TI - Oxytocin-induced desensitization of the oxytocin receptor. AB - OBJECTIVE: The purpose of this study was to characterize the oxytocin-induced desensitization of oxytocin-stimulated rises of intracellular calcium in cultured human myocytes. STUDY DESIGN: Culture lines were begun from biopsy specimens of myometrium that had been obtained from women who underwent low transverse cesarean deliveries. Fluorescence changes of calcium green-1 were used to demonstrate the rises of intracellular free calcium. Cells were exposed to 10 nmol/L oxytocin for 1 to 6 hours before the experimentation, allowed to rest for 10 minutes, and then tested for the fluorescence increases that resulted from exposure to 10 nmol/L oxytocin and micromol/L prostaglandin F(2)(alpha). Subpopulations were defined as type 1 (responded to both oxytocin and prostaglandin F(2)(alpha)), type 2 (responded only to oxytocin), type 3 (responded only to prostaglandin F(2)(alpha)), or type 4 (responded to neither). The distribution of the subpopulations of cells was assessed by the determination of the response of every cell in every experimental run. RESULTS: Pretreatment with oxytocin resulted in a decrease in the percentage of cells that responded to subsequent oxytocin exposure. The decrease was dependent on the duration of oxytocin exposure and was well fit with the Boltzmann sigmoid function. The duration of oxytocin exposure that yielded half-inactivation was 4.2 hours. Without oxytocin pretreatment, the distribution of subpopulations were 37.0% +/- 18.0% (type 1), 23.1% +/- 11.5% (type 2), 12.6% +/- 8.0% (type 3), and 27.3% +/- 22.9% (type 4). After 6 hours of oxytocin pretreatment, the percentage of type 1 and type 2 cells decreased to 2.4% +/- 3.8% and 2.6% +/- 2.4%, respectively, although the percentage of type 3 and type 4 cells increased to 20.4% +/- 18.9% and 74.6% +/- 22.1%, respectively. CONCLUSION: Oxytocin-induced desensitization of myocytes to oxytocin stimulation occurred over a clinically relevant time frame (4.2 hours). Continued responsiveness of the cells to prostaglandin F(2)(alpha) stimulation after 6 hours of oxytocin pretreatment indicated that postreceptor signaling pathways were maintained, which indicates that the oxytocin receptor likely is involved in the mechanism of myocyte desensitization to oxytocin stimulation. PMID- 12592263 TI - Obstetric predictors of placental/umbilical cord blood volume for transplantation. AB - OBJECTIVE: Umbilical cord blood is an effective alternative to bone marrow as a source of hematopoietic stem cells in transplantation. However, the amount of donor blood and the cell content that are collected may be insufficient for engraftment in some adult recipients. This study identifies obstetric factors that affect retrievable placental cord blood volume. STUDY DESIGN: A retrospective analysis of factors that were obtained by direct observation or medical record review that were related to harvested cord blood volume was conducted; the analysis involved 9205 deliveries from mothers who donated placental cord blood through the obstetric services of two New York City hospitals between 1993 and 1999. RESULTS: Obstetric factors that influenced significantly the total volume of blood that was collected were route of delivery, induction of labor, presence of a nuchal cord, infant birth weight, multiple births, placental weight, and duration of labor. The length of the umbilical cord from the venipuncture site and the length of time to cord blood collection also affected the volume that was retrieved. Maternal ethnicity was associated with cord blood yield; Caucasian mothers provided larger quantities than either African American or Asian mothers. CONCLUSION: Our results confirm that the volume of residual placental cord blood that is collected for hematopoietic stem cell transplantation is influenced by several factors, the presence of which predict the likelihood of an adequate collection. Collected volumes can be improved when a longer length of the cord is left with the placenta and when there is a shorter time between the delivery of the placenta and the collection. PMID- 12592264 TI - Fetal plasma stimulates endothelial cell production of cytokines and the family of suppressor of cytokine signaling in umbilical placental vascular disease. AB - OBJECTIVE: We have shown that fetal plasma from pregnancies with placental vascular disease that were identified by an abnormal umbilical artery Doppler study causes endothelial cell activation. We investigated the hypothesis that this would be associated with endothelial cell production of cytokines and their natural regulators, the suppressor of cytokine signaling family. Activation of suppressor of cytokine signaling at the time of cytokine release confirms the fact that cytokine production is occurring in a stimulated cell. STUDY DESIGN: Aliquots from a common culture of human umbilical vein endothelial cells were incubated with fetal plasma from normal pregnancy (n = 29 pregnancies), from umbilical placental vascular disease defined by abnormal umbilical artery Doppler waveforms (n = 38 pregnancies), and from preeclampsia with normal umbilical artery Doppler scans (n = 10 pregnancies). The expression of messenger RNA for the cytokines interleukin-6 and interleukin-8 and the members of suppressor of cytokine signaling family (cytokine-inducible SH2-containing protein, suppressor of cytokine signaling 1, 2, and 3) were assessed by reverse transcriptase polymerase chain reaction. RESULTS: Endothelial cell expression of interleukin-6 messenger RNA (1.94 +/- 0.24 vs 1.31 +/- 0.16) and interleukin-8 messenger RNA (2.62 +/- 0.33 vs 1.64 +/- 0.22) were enhanced in response to incubation with fetal plasma from placental vascular disease in comparison to incubation with fetal plasma from normal pregnancy. The messenger RNA expression of suppressor of cytokine signaling-2 (2.03 +/- 0.23 vs 1.37 +/- 0.16) was up-regulated significantly in placental vascular disease. Differences for cytokine-inducible SH2-containing protein, suppressor of cytokine signaling-1, and suppressor of cytokine signaling-3 were not significant. The expression of cytokines and the suppressor of cytokine signaling family did not differ from normal in the group with maternal preeclampsia and a normal umbilical study. Interestingly, in the umbilical placental vascular disease group, the results were similar in the subgroups, with or without preeclampsia in the mother. CONCLUSION: We have shown that factors that cause endothelial cell injury are present in the fetal circulation in umbilical placental vascular disease. This study is the first report of cytokine production and release and activation of the suppressor of cytokine signaling family by endothelial cells in response to fetal plasma in placental vascular disease. The role of all members of the suppressor of cytokine signaling family in this process must be investigated further. The fact that both the agonist (cytokines) and the antagonist (suppressor of cytokine signaling-2) are produced points to a significant role of endothelial cells in this disease. PMID- 12592265 TI - Hypoxia activates the human placental vascular endothelial growth factor system in vitro and in vivo: up-regulation of vascular endothelial growth factor in clinically relevant hypoxic ischemia in birth asphyxia. AB - OBJECTIVE: We investigated the influence of acute hypoxia on the placental vascular endothelial growth factor system in vitro and in vivo in acute birth asphyxia compared with pregnancies that were complicated by preeclampsia and with healthy control subjects. STUDY DESIGN: Messenger RNA levels for vascular endothelial growth factor, flt-1, and KDR were measured by TaqMan real-time polymerase chain reaction in human placental choriocarcinoma cells (BeWo) that were exposed to hypoxia (1% oxygen, 5% carbon dioxide, 94% nitrogen) and in placental tissue of neonates with birth asphyxia (n = 20), newborn infants of mothers with preeclampsia (n = 20), and gestational age-matched control subjects. Immunhistologically, placental vascular endothelial growth factor protein expression was compared among the groups. RESULTS: In BeWo cells, vascular endothelial growth factor, flt-1 and KDR messenger RNA increased in a time dependent manner in response to hypoxia. In vivo, vascular endothelial growth factor/beta-actin and KDR/beta-actin messenger RNA were significantly higher in placental tissue of newborn infants with severe hypoxic-ischemic encephalopathy than with newborn infants with mild or no hypoxic-ischemic encephalopathy and control subjects. In chronic placental hypoxia (preeclampsia), vascular endothelial growth factor and both receptors were found to be up-regulated. Increased placental vascular endothelial growth factor expression was confirmed by immunohistologic examination. CONCLUSION: The vascular endothelial growth factor system is up-regulated in response to placental hypoxia and is assumed to be a potential early indicator of severe birth asphyxia. PMID- 12592266 TI - Immediate and delayed effects of antenatal corticosteroids on fetal heart rate: a randomized trial that compares betamethasone acetate and phosphate, betamethasone phosphate, and dexamethasone. AB - OBJECTIVE: The purpose of this study was to evaluate the role of different formulations and different administration kinetics of antenatal corticosteroid therapy on fetal heart rate. STUDY DESIGN: One hundred five patients who were at high risk for preterm delivery were assigned randomly to receive two injections of betamethasone acetate + phosphate, four injections of betamethasone phosphate, or four injections of dexamethasone phosphate. Computerized fetal heart rate was recorded daily through day 4. RESULTS: The three formulations did not differ in their effect on fetal heart rate, which varied significantly as a function of the time of injection. During treatment (day 0-day 1), fetal heart rate variability increased (+9%, P <.05), as did the number of fetal movements felt by the mother (+60% at day 0, P <.001). After treatment (day 2-day 3), variability fell significantly (-14%, P <.01), as did accelerations (-35% at day 2, P <.01). No modifications were still detectable on day 4. CONCLUSION: Antenatal corticotherapy is responsible for two different phases of fetal heart rate modifications that do not vary according to the corticosteroid or the dosage regimen. PMID- 12592267 TI - A blueprint for obstetric critical care. AB - OBJECTIVE: The purpose of this study was to describe our 2-year experience with 483 critically ill peripartum women and to propose a blueprint for obstetric critical care. STUDY DESIGN: This was a prospective study to evaluate all admissions to the Obstetric Intermediate Care Unit and obstetric admissions to medical/surgical intensive care units. Our findings are followed by general recommendations for the organization of obstetric critical care. RESULTS: Almost two thirds of the women had obstetric complications that included pregnancy associated hypertension and obstetric hemorrhage. Medical disorders were most common in the other one third of the women. CONCLUSION: An Obstetric Intermediate Care Unit allows for the continuation of care by obstetricians and results in fewer transfers to medical/surgical intensive care units. Patient treatment depends on hospital size and available resources. In most tertiary centers, the critically ill pregnant woman is best cared for by obstetricians in an Obstetric Intermediate Care Unit. In smaller hospitals, transfer to a medical or surgical intensive care unit may be preferable. PMID- 12592268 TI - Cervical light-induced fluorescence in humans decreases throughout gestation and before delivery: Preliminary observations. AB - OBJECTIVE: The purpose of this study was to investigate gestational changes of cervical light-induced fluorescence, an index of cross-linked collagen, and the relationship between light-induced fluorescence and the time-to-delivery interval. STUDY DESIGN: Fifty patients were included in one of two groups. (1) Twenty-one healthy pregnant women without signs of labor underwent repeated cervical light-induced fluorescence measurement during the last trimester. (2) Light-induced fluorescence was measured in 29 patients with signs of labor, and the time from measurement to delivery was noted. Cervical light-induced fluorescence was obtained noninvasively with a prototype instrument that was designed specifically for this purpose. Spearman correlation and the Student t test and receiver-operator characteristics analysis were performed (P <.05). RESULTS: Light-induced fluorescence correlated negatively with gestational age and positively with time-to-delivery interval, was significantly lower in patients who were delivered <24 hours compared with those patients who were delivered >24 hours later, and was predictive of delivery within 24 hours. CONCLUSION: Cervical light-induced fluorescence decreases significantly as gestational age increases. Light-induced fluorescence may be a useful tool to identify patients in whom delivery is imminent. PMID- 12592269 TI - Cohort study of maternal views on future mode of delivery after operative delivery in the second stage of labor. AB - OBJECTIVE: The purpose of this study was to assess maternal views on the future mode of delivery after either previous instrument vaginal delivery or cesarean delivery at full dilatation. STUDY DESIGN: We conducted a prospective cohort study of 393 women with term singleton cephalic pregnancies. RESULTS: More than one half of the cohort intended to have a further pregnancy, with no significant differences between the instrument vaginal delivery and caesarean delivery groups (51% vs 54% before discharge; adjusted odds ratio, 1.04; 95% CI, 0.2, 6.0), and there was little change in maternal views over time. Women were more likely to aim for a future vaginal delivery after an instrument vaginal delivery (79% vs 39% before discharge; adjusted odds ratio, 4.5; 95% CI, 2.2, 9.2), but the proportion decreased over time (68% vs 42% at 1 year; adjusted odds ratio, 3.6; 95% CI, 1.6, 8.1). There were no significant differences in preferred future mode of delivery for women who had a caesarean delivery after a failed instrument delivery and those who were delivered by immediate caesarean delivery. CONCLUSION: A high proportion of women who have had a previous difficult instrument vaginal delivery would still prefer vaginal delivery in a future pregnancy. PMID- 12592270 TI - Mechanical ventilation in an obstetric population: characteristics and delivery rates. AB - OBJECTIVE: The purpose of this study was to describe the characteristics and outcomes of obstetric patients who require mechanical ventilation. STUDY DESIGN: A review was conducted of obstetric patients who required mechanical ventilation and who received care at our institutions between 1990 and 1998. Data that were collected included maternal demographics, medical condition that necessitated ventilation, delivery status, duration of ventilation, onset of parturition while receiving ventilation, mode of delivery, and maternal and early neonatal morbidity or death. RESULTS: Fifty-one women were identified; 43 women(84%) received care in the labor and delivery setting. The most common admission diagnoses were preeclampsia/eclampsia (44%), labor/preterm labor (14%), and pneumonia (12%). Forty-three women (86%) were undelivered on admission (mean gestational age, 31.6 weeks). Delivery occurred in 37 women (86%) during their admission; 24 women (65%) underwent cesarean delivery. Eleven women began labor while receiving ventilation; 6 were delivered vaginally. The maternal mortality rate was 14% (7/51 women), and the perinatal mortality rate was 11% (4/37 fetuses). CONCLUSION: A large number of obstetric patients who receive mechanical ventilation will require delivery because of their condition. Centers that care for such women should form a treatment strategy to coordinate obstetric and medical care for this unique population PMID- 12592271 TI - Myometrial thickness during human labor and immediately post partum. AB - OBJECTIVE: Morphologic studies suggest dramatic, asymmetric uterine growth during pregnancy that is caused by muscle cell hypertrophy. This growth is most marked at the fundus. Our objective was to evaluate sonographically the in vivo changes in myometrial thickness during active labor, second-stage labor, and after delivery. STUDY DESIGN: Abdominal ultrasound scans were performed on 52 term pregnant women to investigate the dynamic changes in myometrial thickness during the active and second stages of labor and immediately after delivery. Twenty-six women (mean +/- SEM gestational age, 39.09 +/- 0.3 weeks) were in active labor (cervical dilatation >4 cm with regular uterine contractions). An additional 26 nonlaboring women (gestational age, 39.8 +/- 0.2 weeks) provided control measurements. The myometrium was defined sonographically as the echo homogeneous layer between the serosa and the decidua. Myometrial thickness was measured at the low segment and mid anterior, fundal, and posterior uterine walls by the same observer. Myometrial thickness was also measured during uterine contractions and after artificial rupture of the amniotic membranes. All laboring women had uncomplicated labor patterns when studied and were delivered spontaneously. RESULTS: The myometrium was significantly thinner during active labor compared with nonlabor at each site studied: midanterior (mean [+/-SEM] myometrial thickness, 5.8 +/- 0.27 vs 8.83 +/- 0.51 mm; t test, P <.001), fundus (mean myometrial thickness, 6.78 +/- 0.32 vs 8.49 +/- 0.35 mm; P =.0015), and posterior (mean myometrial thickness, 6.22 +/- 0.34 vs 8.12 +/- 0.30 mm; P <.001). However, myometrial thickness did not differ among sites within the two groups. The thickness of the low segment was not affected by labor status (nonlabor, 4.68 +/- 0.48 vs labor, 4.66 +/- 0.37 mm; P =.97). Similarly, the myometrial thickness of the anterior uterine wall was unaffected by contractions (no contractions, 5.56 +/- 0.2 vs contractions, 5.68 +/- 0.22 mm; t test, P =.654). There was no change in myometrial thickness measured immediately before and after rupture of the amniotic membranes, despite a significant decrease of the amniotic fluid index. There was significant thickening of the anterior and fundal myometrium during the second stage of labor after the fetal head descended to +3 station by digital examination (anterior, 12.99 +/- 0.60 vs 5.8 +/- 0.27 mm; t test, P <.001; fundus, 10.61 +/- 1.63 vs 6.78 +/- 0.32 mm; t test, P =.04). Valsalva maneuver (pushing) during contractions did not affect myometrial thickness at the fundus (between contractions, 10.61 +/- 1.63 vs pushing, 10.76 +/- 1.95 mm; t test, P =.99). Immediately after delivery, the myometrial thickness at the placental insertion site was the thinnest. After completion of the third stage of labor, the uterine fundus remained significantly thinner than the anterior and posterior walls (fundus, 27.37 +/- 3.5 mm vs anterior, 40.94 +/- 3.5 vs posterior, 42.34 +/ 2.44; one-way analysis of variance, P =.02). CONCLUSION: There is significant and widespread thinning of the myometrium during active labor. Descent of the fetal head during the second stage of labor is associated with a significant relative thickening of the anterior and fundal myometrium. After delivery, the relationship reverses. These findings suggest the directionality of the expulsive force vectors (fundal dominance) is not determined by asymmetric myometrial growth but, rather, may be a function of increased "myometrial mass" that results from increased surface area at the fundus. PMID- 12592272 TI - Comparative efficacy and cost of the prostaglandin analogs dinoprostone and misoprostol as labor preinduction agents. AB - OBJECTIVE: The purpose of this study was to compare the relative efficacy and cost of three commercially available prostaglandin analogs, misoprostol (Cytotec), dinoprostone gel (Prepidil), and dinoprostone insert (Cervidil), as labor preinduction agents. STUDY DESIGN: One-hundred eleven women with an unfavorable cervix who underwent labor induction were assigned randomly to receive either misoprostol 50 microg every 6 hours for two doses, dinoprostone gel 0.5 mg every 6 hours for two doses, or dinoprostone insert 10 mg for one dose intravaginally. Twelve hours later, oxytocin induction was initiated per standardized protocol. Efficacy and cost of the labor preinduction/induction with the study treatments were compared. RESULTS: Mean Bishop score change (+/-SD) over the initial 12-hour interval was significantly greater in the misoprostol group (5.2 +/- 3.1) compared with the dinoprostone insert (3.2 +/- 2.3) or the dinoprostone gel groups (2.2 +/- 1.3, P <.0001). The proportion of women who reached complete dilation (68.4%, 50.0%, 51.4%, respectively; P =.14) and who were delivered (60.5%, 47.4%, 40.0%, respectively; P =.10) within 24 hours of the initiation of induction were not significantly different between the misoprostol, dinoprostone insert, and dinoprostone gel groups. Induction-to-delivery intervals, however, were significantly shorter among women who treated with misoprostol (24.0 +/- 10.8 hours) compared with either the dinoprostone gel (31.6 +/- 13.4 hours) or the dinoprostone insert (32.2 +/- 14.7 hours, P <.05). Overall mean cost per patient that was incurred by labor induction was significantly less for the misoprostol group ($1036.13) compared with the dinoprostone insert group ($1565.72) or the dinoprostone gel group ($1572.92, P <.0001). No significant differences were noted with respect to the mode of delivery or to the adverse maternal/neonatal outcome. CONCLUSION: Misoprostol is more cost-effective than the comparable commercial dinoprostone prostaglandin preparations as an adjuvant to labor induction in women with an unfavorable cervix. PMID- 12592273 TI - Ultrasonographic prediction of term birth weight: how accurate is it? AB - OBJECTIVE: The purpose of this study was to investigate the accuracy of 25 ultrasonic algorithms for the estimation of term fetal weight and to compare these results to an equation that is based on maternal and pregnancy-specific characteristics alone. STUDY DESIGN: Ultrasonography was performed in 82 nondiabetic gravid women at 35 to 41 weeks of gestation. Fetal biparietal diameter, abdominal circumference, head circumference, and femur length were measured. Twenty-five ultrasonic fetal biometric algorithms and an equation that is based only on maternal characteristics were used to predict birth weight. RESULTS: The correlation between predicted and actual birth weight ranged from 0.44 to 0.79 for the ultrasonic algorithms and was 0.60 for the maternal characteristics equation. Ultrasonic algorithms had mean absolute prediction errors that ranged from +/-263 to 646 g (+/-7.5%-18.8%). Accuracy for the maternal characteristics equation was not statistically different from the best performing ultrasonic algorithms (+/-353 g, +/-10.4%). CONCLUSION: Term birth weight estimates that use ultrasonography are generally no more accurate than predictions that are based solely on quantitative assessment of maternal and pregnancy-specific characteristics. PMID- 12592274 TI - Don't ask, don't tell: a change in medical student attitudes after obstetrics/gynecology clerkships toward seeking consent for pelvic examinations on an anesthetized patient. AB - OBJECTIVE: We explore whether the completion of an obstetrics/gynecology clerkship is associated with a decline in the importance that students place on seeking permission from the patient before conducting a pelvic examination while she is anesthetized. STUDY DESIGN: Students at five Philadelphia area medical schools (n = 401 students) were asked how important it would be for a patient to be told that a medical student will perform a pelvic examination while she is anesthetized. We examined associations between the completion of an obstetrics/gynecology clerkship and attitudes toward consent with the use of linear regression to adjust for gender and the total amount of clerkship experience. RESULTS: After the data were controlled for gender and the total number of clerkships that had been completed, we found that students who had completed an obstetrics/gynecology clerkship thought that consent was significantly less important than did those students who had not completed a clerkship (P =.01). CONCLUSION: To avoid this decline in attitudes toward seeking consent, clerkship directors should ensure that students perform examinations only after patients have given consent explicitly. PMID- 12592275 TI - The relationship of race to women's use of health information resources. AB - OBJECTIVE: The purpose of this study was to examine, among the general public, the independent effect of race on women's use of health information resources. STUDY DESIGN: A population-based random-digit dialing survey of adult women, aged 18 to 64 years, was conducted between October 1999 and January 2000. Subjects included 509 women (341 white women, 135 black women, and 33 women of other races). The response rate was 66%. The main outcome variable was the use of health information resources (print health or news media, broadcast media, computer resources [Internet], health organizations, organized health events). Logistic regression was used to determine the independent effect of race/ethnicity on the use of different information resources, with an adjustment for age, income, education, and marital status. RESULTS: After the adjustment for socioeconomic factors, black women had <50% odds of using print news media (odds ratio, 0.5; 95% CI, 0.4-0.8), <60% odds of using computer-based resources (odds ratio, 0.4; 95% CI, 0.2-0.6), and <70% odds of using health policy organizations (odds ratio, 0.3; 95% CI, 0.2-0.7), compared with white women. CONCLUSION: There is a large racial disparity in women's use of health information resources. Traditional sources that are used to provide patient information may not be effective in certain populations. PMID- 12592276 TI - Vaginal sonography and cervical incompetence. AB - Uterine contractions, decidual activation, and cervical competence comprise the fundamental components in contemporary models of the spontaneous preterm birth syndrome, but their relative importance and interactive pathways remain poorly defined. Moreover, the traditional concept that the cervix is either competent or incompetent has been challenged because cervical competence more likely functions along a biologic continuum. Cervical incompetence is a clinical diagnosis characterized by recurrent painless dilation and spontaneous midtrimester birth. Although the efficacy of cerclage for cervical incompetence has never been fully confirmed in randomized clinical trials, the role of cerclage has been expanded to include women with "risk factors" for spontaneous preterm birth or nonreassuring sonographic cervical findings in the mid trimester. Evidence from randomized trials suggests that cerclage has limited efficacy in women with risk factors for cervical incompetence. Results of both retrospective cohort series and randomized trials of cerclage in women with shortened cervical length are inconclusive. We believe that women with a prior early spontaneous preterm birth and shortened midtrimester cervical length represent an ideal population for the conduct of a randomized trial of cerclage, which is currently underway. PMID- 12592277 TI - Biochemical screening for Down syndrome in renal failure. PMID- 12592279 TI - Regarding the PEACH randomized trial. PMID- 12592280 TI - Effectiveness of inpatient and outpatient treatment strategies for women with pelvic inflammatory disease. PMID- 12592281 TI - The PEACH study. PMID- 12592284 TI - Five-year experience with midtrimester amniocentesis performed by a single group of obstetrician-gynecologists at a community hospital. PMID- 12592285 TI - Impact of single- or double-layer closure on uterine rupture. PMID- 12592287 TI - Impact of single-or double-layer closure on uterine rupture. PMID- 12592289 TI - Postpartum hemorrhage mistakes. PMID- 12592292 TI - 1. Overview of the immune response. AB - Host defense against pathogenic microbes requires dramatically different responses, depending on the character of the pathogen and on the tissue under attack. Central to the immune system's ability to mobilize a response to an invading pathogen is its ability to distinguish self from nonself. The host has evolved both innate and adaptive mechanisms to respond to and eliminate pathogenic microbes. Both of these mechanisms include self-nonself discrimination. This overview describes key mechanisms used by the immune system to respond to invading microbes and identifies settings in which disturbed immune function exacerbates tissue injury. PMID- 12592293 TI - 2. Cytokines and chemokines. AB - Cytokines and chemokines are redundant secreted proteins with growth, differentiation, and activation functions that regulate and determine the nature of immune responses and control immune cell trafficking and the cellular arrangement of immune organs. Which cytokines are produced in response to an immune insult determines initially whether an immune response develops and subsequently whether that response is cytotoxic, humoral, cell-mediated, or allergic. A cascade of responses can be seen in response to cytokines, and often several cytokines are required to synergize to express optimal function. An additional confounding variable in dissecting cytokine function is that each cytokine may have a completely different function, depending on the cellular source, target, and, most important, specific phase of the immune response during which it is presented. Numerous cytokines have both proinflammatory and anti inflammatory potential; which activity is observed depends on the immune cells present and their state of responsiveness to the cytokine. For this chapter, cytokines are grouped according to those that are mononuclear phagocytic-derived or T-lymphocytic-derived; that mediate cytotoxic (antiviral and anticancer), humoral, cell-mediated, or allergic immunity; and that are immunosuppressive. The biology of chemokines are then reviewed, grouped by family. PMID- 12592294 TI - 3. Lymphocytes. AB - The fundamental task of the immune system is to defend "self" from "nonself." Lymphocytes are the primary cells of the immune system that developed one of the most sophisticated and comprehensive defense mechanisms in the biological system. T cells play a central role in orchestrating the immune response. Further, they are instrumental in eliminating intracellular pathogens (viruses, some bacteria) through the generation of cytotoxic T cells. B cells defend against extracellular pathogens by producing antibodies. Natural killer cells are an important component of innate immunity. Dendritic cells play a key role in initiating the immune response by presenting foreign antigens to T cells. The interaction among T cells, B cells, dendritic cells, and natural killer cells constitute the fundamental defense network of the host. The failure of any of these components severely jeopardizes the integrity of the immune system and its ability to mount the most appropriate immune response. PMID- 12592295 TI - 4. IgE, mast cells, basophils, and eosinophils. AB - IgE, mast cells, basophils, and eosinophils constitute essential elements in allergic inflammation. Allergen-specific IgE, synthesized in response to allergens in the environment and in susceptible individuals, becomes fixed to high-affinity receptors on cellular membranes, especially of mast cells and basophils. If these receptor-bound IgE molecules are aggregated on reexposure to specific allergen, these mast cells and basophils produce mediators that result in the allergic response. Principal among the cells drawn to sites of mediator release is the eosinophil. PMID- 12592296 TI - 5. Genetics of hypersensitivity. AB - Genetics provides the basis for the host response to a variety of environmental factors that can play a role in the generation of complex genetic diseases, such as asthma and atopy. An understanding of the genetic bases for these conditions is therefore essential to understand their pathophysiology. Studies of the genetics of asthma and atopy have suffered from several daunting challenges. These include the recognition that these are conditions caused by numerous genes, with each gene assuming variable roles in different individuals. In addition, each gene presumably contributes only a small percentage to a given individual's genetic risk of asthma. This has led to the current situation, in which studies often demonstrate a lack of replication that can be explained by their being insufficiently powered. Furthermore, the pathophysiologies of asthma and atopy are incompletely understood, and the lack of clearly defined phenotypes also contributes to the inadequacies of the current literature. Nonetheless, regions of the human genome have been reproducibly associated with asthma and atopy. These regions have undergone intense study, and many genetic variants within them have been implicated as asthma and allergy genes. In addition, through candidate gene approaches, several genetic polymorphisms have been convincingly linked to increased risks for the development of asthma and atopy. Many of these genes are associated with alterations in responsiveness to therapeutic agents used in the treatment of these conditions. These genetic studies have an exciting potential for individually tailoring the therapeutic regimen to a given subject's genotype. It is to be hoped that they will also define new targets for the next generation of asthma and allergy therapeutic agents. PMID- 12592297 TI - 6. Asthma. AB - The increasing incidence and prevalence of asthma in many parts of the world continue to make it a global health concern. The heterogeneous nature of the clinical manifestations and therapeutic responses of asthma in both adult and pediatric patients indicate that it may be more of a syndrome rather than a specific disease entity. Numerous triggering factors including viral infections, allergen and irritant exposure, and exercise, among others, complicate both the acute and chronic treatment of asthma. Therapeutic intervention has focused on the appreciation that airway obstruction in asthma is composed of both bronchial smooth muscle spasm and variable degrees of airway inflammation characterized by edema, mucus secretion, and the influx of a variety of inflammatory cells. The presence of only partial reversibility of airflow obstruction in some patients indicates that structural remodeling of the airways may also occur over time. Choosing appropriate medications depends on the disease severity (intermittent, mild persistent, moderate persistent, severe persistent), extent of reversibility, both acutely and chronically, patterns of disease activity (exacerbations related to viruses, allergens, exercise, etc), and the age of onset (infancy, childhood, adulthood). PMID- 12592298 TI - 7. Rhinitis and sinusitis. AB - Rhinitis and sinusitis are prevalent medical conditions that are often associated with each other and may result in significant morbidity and medical costs. They can cause systemic symptoms, decrease quality of life, and result in reduced workplace productivity and missed school days. Appropriate management of rhinitis or sinusitis may be an important component in effective management of coexisting or complicating conditions, such as asthma, allergic conjunctivitis, or chronic otitis media. Rhinitis may be caused by allergic, non-allergic, infectious, hormonal, occupational, and other factors. Defining the basis for rhinitis in an individual is important in selection of therapeutic options. Rhinitis and sinusitis may be difficult to distinguish from each other on the basis of history alone. Although most acute upper respiratory infections are viral and do not require antibiotic treatment, persistence of symptoms for > or =7 days makes acute bacterial sinusitis more likely and antibiotics an appropriate consideration. Radiographic imaging is not required for diagnosis of acute, uncomplicated sinusitis, although CT scans are indicated in evaluation of suspected chronic sinusitis or treatment failures. Chronic sinusitis may have an infectious or non-infectious basis. Underlying disorders that predispose to chronic sinusitis should be identified and treated as part of the treatment of chronic sinusitis. PMID- 12592299 TI - 8. Occupational asthma and allergies. AB - A diversity of airborne dusts, gases, fumes, and vapors can cause dose-related symptoms in individuals exposed in the workplace. More than 250 chemicals have been incriminated as a cause of occupational asthma (OA). The prevalence of OA ranges from 2% to 6% of the asthmatic population. Predisposing factors facilitating the development of OA include the work environment, climatic conditions, genetic proclivities, tobacco and recreational drug use, respiratory infection, and bronchial hyperresponsiveness. Pathogenetically, new-onset OA may be immunologic or nonimmunologic in origin. The immunologic variants are usually caused by high molecular-weight allergens such as grain dust and animal or fish protein. Symptoms ensue after a latent period of months to years. Nonimmunologic OA can be precipitated by a brief, high-level exposure to a potent irritant. Symptoms occur immediately or within a few hours of the exposure. In either instance, once the diagnosis is established, the worker should be removed from the workplace. If the diagnosis is made in a timely fashion, most workers experience improvement. Prevention is the best therapeutic intervention. PMID- 12592300 TI - 9. Food allergy. AB - Food allergies affect as many as 6% of young children, most of whom "outgrow" the sensitivity, and about 2% of the general population. Although any food may provoke a reaction, relatively few foods are responsible for the vast majority of food allergic reactions: milk, egg, peanuts, tree nuts, fish, and shellfish. Many of these food allergens have been characterized at a molecular level, which has increased our understanding of the immunopathogenesis of many responses and may soon lead to novel immunotherapeutic approaches. Food allergic reactions are responsible for a variety of symptoms involving the skin, gastrointestinal tract, and respiratory tract and may be due to IgE-mediated and non-IgE-mediated mechanisms. A systematic approach including history, laboratory studies, elimination diets, and often food challenges will lead to the correct diagnosis. Currently, management of food allergies consists of educating the patient to avoid ingesting the responsible allergen and to initiate therapy in case of an unintended ingestion. PMID- 12592301 TI - 10. Drug allergy. AB - Adverse drug reactions are common, but only 6% to 10% are immunologically mediated. Unlike most adverse drug reactions, allergic drug reactions are unpredictable. Whereas some drug-induced allergic reactions may be easily classified into one of the four Gell and Coombs hypersensitivity categories, many others that appear to have an immunologic component cannot be classified because of our lack of mechanistic information. Theoretically, any drug can induce an immune response. However, some drugs are more likely to elicit clinically relevant immune responses than are others. Drugs in this category include antimicrobial drugs, anticonvulsants, chemotherapeutic agents, heparin, insulin, protamine, and biologic response modifiers. After a drug-disease connection is established, it must be determined whether the reaction was immunologically mediated. Subsequently, confirmatory tests, if available, should be used to determine the allergic status of the patient. If these tests are not available, a graded challenge or desensitization may be considered, depending on the type of clinical reaction previously demonstrated and the need for drug readministration. Education of the patient and primary care physician is an important component of patient management. PMID- 12592302 TI - 11. Allergic and immunologic diseases of the skin. AB - Many skin diseases have an inflammatory or immune component, and anti inflammatory drugs comprise a major portion of a dermatologist's therapeutic armamentarium. Although causes of most of these diseases remain obscure, mechanisms of lesion formation and explanations for symptoms are increasingly well documented. These developments, coupled with the expected availability of novel selective immunomodulatory agents, herald a new era for immunodermatology. Patients with psoriasis, allergic contact dermatitis, atopic dermatitis, urticaria, and autoantibody-mediated blistering diseases are among those who are likely to benefit from advances in the understanding of disease pathogenesis and the emergence of immunotherapeutics. PMID- 12592303 TI - 12. Primary immunodeficiency diseases. AB - Although primary immunodeficiency disorders are relatively rare, intensive investigation of these disorders has yielded a great wealth of understanding of basic immunologic mechanisms in host defense, inflammation, and autoimmunity. These advances have led to important developments for the treatment not only of the primary immunodeficiencies but also for patients with secondary immunocompromised states, autoimmune disorders, hypersensitivity, graft rejection, and graft versus host disease. Correction of a form of severe combined immunodeficiency represents the first true success of human gene therapy. This review introduces the major clinical manifestations of primary immunodeficiency disorders, along with descriptions of essential elements of the pathophysiology of those disorders that have been defined at the molecular level. Key concepts in treatment are also presented. It is critical for the practicing primary care provider and allergist to maintain an index of suspicion for immunodeficiency. Early diagnosis offers the best opportunity for reduced morbidity and survival and is critical for accurate genetic counseling. PMID- 12592304 TI - 13. HIV-1 infection. AB - This review is intended to provide a fundamental perspective on the dynamic interplay between HIV-1 and the immune system, an essential aspect in defining the pathogenesis and treatment of AIDS. HIV-1 infection, the cause of AIDS, is a worldwide pandemic with enormous adverse heath and economic implications, particularly in the developing world. This bloodborne and sexually transmitted disease, which evolved from simian immunodeficiency virus, infects and replicates in helper T cells and macrophages and utilizes CD4 and a chemokine coreceptor for entry. Immune deficiency occurs as a result of virally induced attrition of CD4 T cells, resulting in the development of opportunistic infections and malignancy. Prophylaxis against opportunistic infections is required according to the extent of immune deficiency. HIV-specific immunity can control viral replication and delay disease progression but does not clear infection. Antiretroviral treatment consists of inhibitors that target for viral entry, reverse transcriptase, and viral protease. Therapy can control viral replication, restore immunity, and delay disease progression, but it cannot eliminate infection. Thus chronic infection persists even in treated patients. Antiretroviral drugs have been highly effective in preventing mother-to-child transmission and for postexposure prophylaxis. Several novel vaccines in development hold promise for either effective infection prevention or attenuation of disease progression. PMID- 12592305 TI - 14. Immunologic rheumatic disorders. AB - We provide the basics for the clinician who might be called on to consider the diagnosis of diseases such as systemic lupus erythematosus (SLE) or rheumatoid arthritis (RA) in their practice. We will emphasize clinical recognition and first-line laboratory testing. Only characteristics of the classic rheumatic inflammatory diseases, RA, SLE, Sjogren syndrome, scleroderma, and dermatomyositis/polymyositis, will be covered. In the past decade, RA is the only disease for which treatment has substantially improved. The treatment of RA has been revolutionized by the use of methotrexate and, more recently, tumor necrosis factor inhibitors. The goal of RA treatment today is to induce a complete remission as early as possible in the disease process, with the mantra being "elimination of synovitis equals elimination of joint destruction." The hope is that if the major mediators of Sjogren syndrome or SLE or scleroderma can be identified and then blocked, as in the example of tumor necrosis factor inhibitors in RA, more specific treatments will become available. Thus, RA has become an excellent model of this evolving paradigm. Through the identification of major mediators in its pathogenesis, novel and highly efficacious therapeutic agents have been developed. PMID- 12592306 TI - 15. Vasculitis. AB - Vasculitis is histologically defined by the presence of blood vessel inflammation. It can be observed in a wide variety of settings, either occurring secondarily to another process or as the pathologic foundation of a primary vasculitic disease. The primary systemic vasculitides comprise a broad group of disease entities that are uniquely identified by the nature of their clinical, histopathologic, or therapeutic characteristics. Individual diseases often predominantly affect blood vessels of a particular size, the pattern of which influences their clinical manifestations and has been used in their classification. The vasculitides span a wide range of disease severity, extending from illnesses that rarely produce death to those almost universally fatal before the introduction of effective therapy. Immunosuppressive and cytotoxic agents are used to treat many vasculitic diseases. Although such approaches can be effective, long-term treatment may be complicated by chronic sequelae from organ damage, disease relapses, and medication side effects. Recent investigations have focused on understanding the pathophysiology of these diseases, which may lead to more efficacious and less toxic therapeutic options. PMID- 12592307 TI - 16. Immunologic lung disease. AB - This review summarizes the recent advances regarding pathogenesis, diagnosis, and treatment of immunological diseases of the lung. Rather than attempt a comprehensive analysis, we have focused on selected diseases that are of particular relevance to the practicing physician, and the material has been organized according to the dominant immunologic mechanisms underlying the disease. Because of the redundancy that characterizes the mammalian immune repertoire, this system of classification inevitably produces overlap but facilitates acquisition of what is otherwise a disparate collection of facts. The principal lung immunologic mechanisms are most broadly classified as innate or adaptive immune processes. Innate immunity includes neutrophils and complement that are important in diseases, such as pneumonia and the acute respiratory distress syndrome. Adaptive immunity involves T and B cells capable of recognizing discrete antigens. T(H)1- and T(H)2-dependent adaptive immune responses underlie some of the most common and important of lung diseases, including tuberculosis and asthma, respectively. Other important immunopathologic processes include granulomatous inflammation that characterizes sarcoidosis and Churg-Strauss vasculitis, and autoimmunity, which is characteristic of antiglomerular basement membrane disease and others. PMID- 12592308 TI - 17. Immunologic endocrine disorders. AB - Immune-mediated tissue destruction or disregulation is the cause of multiple common, as well as rare, endocrine disorders including type 1 diabetes, Graves' disease, Hashimoto thyroiditis, and Addison's disease. Each of these disorders can be divided into a series of stages beginning with genetic susceptibility, environmental triggering events, and active autoimmunity, followed by metabolic abnormalities with overt disease. Common genetic susceptibility is suggested by the clustering of a series of disorders in the same individual and his or her family. A major portion of the genetic susceptibility lies in the HLA region, but for several disorders, mutation of transcription factors underlies disease susceptibility (eg, X-linked polyendocrinopathy, immune deficiency and diarrhea, and autoimmune polyendocrine syndrome type 1). With improving immunogenetic and pathogenic understanding, type 1A diabetes is now predictable, and excellent autoantibody screening assays are available. This knowledge, combined with studies in animal models, has led to trials for the prevention of diabetes. In addition, aberrant immunologic reactions (eg, insulin autoantibodies after insulin therapy, Graves' disease after monoclonal anti-T-cell therapy in multiple sclerosis) can complicate standard and experimental therapies. We therefore believe that an understanding of the immunogenetics and immunopathogenesis of endocrine disorders can aid in the prevention of morbidity and mortality for these related diseases. PMID- 12592309 TI - 18. Immune-mediated renal disease. AB - Immune-mediated renal diseases can be classified by the clinical syndromes they produce, by the attendant renal pathology, or by the dominant immune effector mechanism of renal injury. The major clinical syndromes produced by immune mediated renal injury include the nephrotic syndrome, the nephritic syndrome, rapidly progressive glomerulonephritis, and acute renal failure. The notion of clinical syndromes facilitates diagnosis and treatment, but does not accurately define disease pathogenesis. In this summary, we discuss pathologically defined immune-mediated renal diseases under the clinical syndrome with which they are most frequently associated. There is overlap between the clinical syndromes, but the syndromes provide a useful framework. Relevant information regarding the proposed pathogenesis of disease entities is included under specific disease entities. PMID- 12592310 TI - 19. Immunologic, gastroenterologic, and hepatobiliary disorders. AB - A number of gastrointestinal and hepatobiliary disorders are thought to have an immunologic basis; however, the level of understanding of immunopathogenesis varies widely in these disparate conditions. For some, such as celiac disease, both important genetic and environmental determinants have been identified as well as a specific treatment. For others, such as pernicious anemia and inflammatory bowel disease, animal models have provided important research advances. Two conditions, inflammatory bowel disease and sclerosing cholangitis, are strongly associated. Characteristic immunologic features of these diseases are critical for diagnosis. Although the majority of conditions are currently treated with nonselective anti-inflammatory and immunosuppressive agents, recent research has introduced novel biological agents, exemplified by the successful use of infliximab for Crohn's disease. PMID- 12592311 TI - 20. Immunologic neuromuscular disorders. AB - Immune-mediated disorders of each of the structural subdivisions of the nervous and neuromuscular system have been described. Despite the immune privilege of the central nervous system, and to a lesser extent the peripheral nervous system, immune dysregulation is not uncommon. Environmental, genetic, and immunologic factors have been postulated to be involved in the development of these disorders. Major immune-mediated neurologic diseases of the central nervous system include multiple sclerosis and acute disseminated encephalomyelitis. Immune-mediated diseases of the peripheral nervous system include myasthenia gravis, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, idiopathic polymyositis, dermatomyositis, and inclusion body myositis. Some of these disorders, such as myasthenia gravis and certain forms of acute inflammatory demyelinating polyneuropathy, are clearly autoimmune in nature, whereas the immune system plays an important role in pathogenesis in others. Understanding the immune mechanisms of disease and uncovering potential therapeutic targets are essential for the design of new treatments. The epidemiology, pathogenesis, diagnostic criteria, and current therapeutic approaches to the major neuroimmunologic diseases are reviewed. PMID- 12592312 TI - 21. Immunohematologic disorders. AB - Immunohematologic disorders are a broad group of entities in which hematologic diseases, usually cytopenias, are caused by immune reactions. These reactions lead to the development of hemolytic anemia, thrombocytopenia, or neutropenia, either separately or in combination. Common underlying mechanisms include immunodeficiency, systemic autoimmunity, and drug-induced reactions. Transplacental transfer of maternal alloantibodies can lead to fetal and neonatal cytopenias. Other immune reactions include antibodies to clotting factors, which result in both thrombosis and hemorrhage, and hemolytic and nonhemolytic reactions during transfusion of blood products. PMID- 12592313 TI - 22. Immune responses to malignancies. AB - Immune responses to tumor-associated antigens exist in tumor-bearing hosts but are usually not successful in eliminating malignant cells or preventing the development of metastases. Patients with cancer generate robust immune responses to infectious agents (bacteria and viruses) perceived as a "danger signal" but only ineffective, weak responses to tumor-associated antigens, which are considered as "self." This fundamental difference in responses to self versus non self is further magnified by the ability of tumors to subvert the host immune system. Tumors induce dysfunction, as well as apoptosis in CD8(+) antitumor effector cells. The escape of tumors from immune cells is mediated by several distinct molecular mechanisms. Insights into these mechanisms and more effective control of tumor-orchestrated immune dysfunction are needed. Novel strategies for immunotherapy of cancer must address protection and survival of antitumor effector cells in the tumor microenvironment. PMID- 12592314 TI - 23. Clinical laboratory assessment of IgE-dependent hypersensitivity. AB - This chapter reviews clinical and laboratory analyses that aid in the diagnosis and management of human allergic (IgE-dependent) diseases. The diagnostic algorithm for immediate-type hypersensitivity begins with a thorough clinical history and physical examination. Once signs and symptoms compatible with an allergic disorder have been identified, a skin test and/or blood test for allergen-specific IgE antibodies may serve as primary confirmation to strengthen the diagnosis. Puncture and intradermal skin testing provide a biologically relevant immediate-type hypersensitivity response in the skin, with resultant wheal and flare reactions within 15 minutes of allergen application. Bleeding, dermatographism, and antihistamines may confound the quality of the skin test. Allergen-specific IgE antibody may also be detected in the blood using a radioallergosorbent test (RAST). Nonisotopic "second-generation" RAST-type assays have evolved to provide more quantitative, sensitive, precise IgE antibody results. In vivo provocation tests may serve as secondary confirmatory tests when the clinical history is discordant with a primary IgE antibody test result. The multiallergen screen is a qualitative RAST-type assay that detects specific IgE antibody to approximately 15 allergens that evoke a large majority of aeroallergen or food-related allergic disorders. Other useful serological assays performed in the diagnostic allergy laboratory include total serum IgE, Hymenoptera venom-specific IgG antibody, IgG precipitins for organic dusts, mast cell tryptases, and the venom RAST inhibition test. Above all, in vivo or laboratory confirmatory test results that are inconsistent with the clinical history should be repeated as for any laboratory assessment. PMID- 12592316 TI - 25. Immunotherapy of allergic disease. AB - Specific immunotherapy involves the administration of allergen extracts to achieve clinical tolerance of the allergens which cause symptoms in patients with allergic conditions. Immunotherapy has been shown to be effective in patients with mild forms of allergic disease, and also in those who do not respond well to standard drug therapy. Recent studies suggest that specific immunotherapy may also modify the course of allergic disease, by reducing the risk of developing new allergic sensitizations, and also inhibiting the development of clinical asthma in children treated for allergic rhinitis. Specific immunotherapy remains the treatment of choice for patients with systemic allergic reactions to wasp and bee stings. The precise mechanisms responsible for the beneficial effects of SIT remain a matter of research and debate. An effect on regulatory T cells seems most probable, associated with switching of allergen-specific B cells towards IgG4 production. Few direct comparisons of specific immunotherapy and drug therapy have been made. Existing data suggest that the effects of specific immunotherapy take longer to come on, but once established, specific immunotherapy will give long-lasting relief of allergic symptoms, whereas the benefits of drugs only last as long as they are continued. PMID- 12592315 TI - 24. Clinical and laboratory assessment of immunity. AB - Clinicians order specific laboratory tests and interpret laboratory data that will be useful in establishing a diagnosis of immunodeficiency, either inherited (primary) immunodeficiencies or acquired (secondary) immunodeficiencies. The clinical immunologic evaluation of patients for immunodeficiency proceeds in an orderly fashion, from screening tests to sophisticated tests. The medical history and physical examination of the patient frequently provide clues guiding the level of entry into this testing program. Specific tests have been designed to screen for the four basic mechanisms of host defense: Antibody, T cell, phagocyte, and complement. The clinical immunology laboratory is a powerful adjunct to the clinician in the initial evaluation of immunodeficiency disorders and in the design of more sophisticated testing for selected patients. PMID- 12592317 TI - 26. Immunomodulation and immunotherapy: drugs, cytokines, cytokine receptors, and antibodies. AB - The preceding chapters in this primer have provided an overview of the immune response that serves as a background for understanding potential sites for immune modulation and immunotherapy. A number of soluble growth and activation factors are released from various cell populations involved in the immune response. They play vital roles in the initiation, propagation, and regulation of immunologic responses. Pharmacologic immunomodulators include suppressive and stimulatory agents. Immunosuppressive therapies include antimetabolites, cytotoxic drugs, radiation, adrenocortical glucocorticosteroids, immunophilins, and therapeutic antibodies. The field of clinical immunostimulation is emerging as an important therapeutic modality for a number of immunodeficiency diseases, chronic viral infections, and cancer. These compounds will be discussed in terms of general principles, molecular targets, major indications, and adverse effects. PMID- 12592318 TI - 27. Transplantation immunology: organ and bone marrow. AB - The discovery of the human MHC in 1967 launched the field of organ and tissue transplantation. More than 800,000 such transplants have been performed during this time. Although matching of donor and recipient for MHC antigens was shown to be of great importance and continues to be so, the development of pharmacologic agents and antilymphocyte antibodies that interfere with the process of graft rejection has had a crucial role in the success of organ transplantation during the past 2 decades. Enormous progress has been made in understanding the immunologic mechanisms of graft rejection and of graft-versus-host disease. The roles of antibodies, antigen-presenting cells, helper and cytotoxic T cells, immune cell surface molecules, and signaling mechanisms and the cytokines they release have been clarified. This understanding is leading to the development of newer immunosuppressive agents targeting various components of the rejection process. Combinations of these agents work synergistically, leading to lower doses and reduced toxicity. Similarly, the development of effective T-cell depletion techniques has been of great importance for bone marrow transplantation when an HLA-identical sibling is not available. The major obstacle to the performance of solid organ transplantation currently is the shortage of donor organs. PMID- 12592319 TI - 28. Embryonic and adult stem cell therapy. AB - Stem cells are characterized by the ability to remain undifferentiated and to self-renew. Embryonic stem cells derived from blastocysts are pluripotent (able to differentiate into many cell types). Adult stem cells, which were traditionally thought to be monopotent multipotent, or tissue restricted, have recently also been shown to have pluripotent properties. Adult bone marrow stem cells have been shown to be capable of differentiating into skeletal muscle, brain microglia and astroglia, and hepatocytes. Stem cell lines derived from both embryonic stem and embryonic germ cells (from the embryonic gonadal ridge) are pluripotent and capable of self-renewal for long periods. Therefore embryonic stem and germ cells have been widely investigated for their potential to cure diseases by repairing or replacing damaged cells and tissues. Studies in animal models have shown that transplantation of fetal, embryonic stem, or embryonic germ cells may be able to treat some chronic diseases. In this review, we highlight recent developments in the use of stem cells as therapeutic agents for three such diseases: Diabetes, Parkinson disease, and congestive heart failure. We also discuss the potential use of stem cells as gene therapy delivery cells and the scientific and ethical issues that arise with the use of human stem cells. PMID- 12592321 TI - 30. Defining the spectrum of clinical immunology. AB - Historically, in the development of the subspecialty of clinical immunology, loosely connected groups of transplanters, endocrinologists, gastroenterologists, and many others have existed, each practicing a special form of immunology pertinent to their area of interest. The diversity of interests and separation of efforts, and the absence of educational initiatives in this field, have conspired to prevent most clinical immunologists from receiving the education required and the recognition appropriate for this subspecialty of medicine. What is proposed here is an attempt to collect the wide spectrum and isolated components of clinical immunology-specific practices into a unifying concept centered around four themes that define the field of clinical immunology better than traditional organ or disease based specialties: (1) immunoregulation, (2) immunotherapy, (3) immunogenetics, and (4) immunodiagnostics. This definition of the themes of clinical immunology provides a cross-disciplinary approach to understanding the present spectrum of interests of clinical immunologists and easily incorporates new interests, such as host defense, as the practice of clinical immunology evolves. In the future, through the use of a thematic approach to the training of clinical immunologists, a new, more organized mechanism may evolve for defining this specialty, educating the members, and providing a common pathway of certification. PMID- 12592320 TI - 29. Immunization. AB - The medical dictionary defines immunization as the "protection of susceptible individuals from communicable diseases by the administration of a living modified agent, a suspension of killed organisms, or an inactivated toxin." This elegant description can be expanded to include twenty-first century approaches to immunization that include recombinant technology, reassortment virus techniques, live vectors, DNA vaccines, and the expansion of the field to encompass noncommunicable diseases such as Alzheimer's disease, autoimmunity, and tumor immunogenetics. Integral to the success of immunization is our knowledge of the immune system's memory of antigens, yet our understanding of this fundamental feature remains limited. On a global scale, communicable diseases remain the number-one cause of morbidity and mortality; hence Jenner's pioneering work with its birth in 1796 still has a challenging and exciting future. PMID- 12592322 TI - 31. Assessment of clinical competence of the allergist/immunologist. AB - The assessment of physician performance is increasingly central to the practice of medicine. Participation in such assessment is a manifestation of adherence by the physician to the highest standards of professionalism. The goal of physician assessment is to assure patients and their families, the public, fellow physicians, and ourselves that we are continuing to perform at the highest levels possible. Traditionally, trainees have been assessed by their program directors, and the methods for such assessment are in place and have, in some cases, been validated against outcomes. Assessment of practicing physicians is a newer concept and arose as a response to the needs of physicians to demonstrate accountability and the demands of the public and regulatory bodies. The leadership of the medical profession and the certifying boards are committed to effective and appropriate assessment and are in the process of implementing physician assessments. To do so, multiple innovative and exciting new evaluative tools are under development. This review discusses the history, background, and current state of the art in this arena. PMID- 12592323 TI - A pediatric approach to the WHO classification of myelodysplastic and myeloproliferative diseases. AB - Myelodysplastic and myeloproliferative disorders are rare in childhood and there is no widely accepted system for their diagnosis and classification. We propose minimal diagnostic criteria and a simple classification scheme which, while based on accepted morphological features and conforming with the recent suggestions of the WHO, allows for the special problems of myelodysplastic diseases in children. The classification recognizes three major diagnostic groups: (1) juvenile myelomonocytic leukemia (JMML), previously named chronic myelomonocytic leukemia (CMML) or juvenile chronic myeloid leukemia (JCML); (2) myeloid leukemia of Down syndrome, a disease with distinct clinical and biological features, encompassing both MDS and AML occurring in Down syndrome; and (3) MDS occurring both de novo and as a complication of previous therapy or pre-existing bone marrow disorder (secondary MDS). The main subtypes of MDS are refractory cytopenia (RC) and refractory anemia with excess of blasts (RAEB). It is suggested retaining the subtype of RAEB-T with 20-30% blasts in the marrow until more data are available. Cytogenetics and serial assessments of the patients are essential adjuncts to morphology both in diagnosis and classification. PMID- 12592324 TI - The interaction of the Bcr-Abl tyrosine kinase with the Src kinase Hck is mediated by multiple binding domains. AB - Bcr-Abl is found in more than 95% of cases with CML. The mechanism of Bcr-Abl induced transformation is not fully understood. Bcr-Abl is a constitutively active tyrosine kinase with transforming capacity for hematopoietic cells. We demonstrated recently that the Src kinase Hck interacts directly with Bcr-Abl by a kinase-independent mechanism. Moreover, the inhibition of the Hck kinase seems to block some of the transforming effects of Bcr-Abl. To identify the binding domains mediating this interaction of Hck with Bcr-Abl, we co-expressed different plasmid and baculovirus vectors containing mutants or single domains of Bcr-Abl and/or Hck in COS7 and Sf9 cells. At least four independent binding regions for Hck were identified in Bcr-Abl, one in Bcr, one in the region comprising the SH3 and SH2 domain of Abl, one in the SH1 domain of Abl, and one in the C-terminal domain of Abl. In the Hck kinase, deletion of the SH2 and/or the SH3 region abolished binding to Bcr-Abl. In contrast, deletion of the Hck SH1 domain enhanced binding of Hck to Abl and Bcr-Abl. In conclusion, the results indicate that the interaction of Bcr-Abl with Hck is mediated by a novel, complex mechanism that involves multiple domains of Bcr-Abl and the SH2 and SH3 domains of Hck. PMID- 12592325 TI - Imatinib mesylate (STI571) in preparation for allogeneic hematopoietic stem cell transplantation and donor lymphocyte infusions in patients with Philadelphia positive acute leukemias. AB - Chronic myeloid leukemia in blast crisis (BC) and Philadelphia chromosome positive acute lymphoblastic leukemia (Ph(+) ALL) are associated with extremely poor outcome. Allogeneic transplantation during BC or active leukemia is most often unsuccessful due to high-rates of both treatment-related complications and relapse. Long-term results are significantly better if a second chronic phase or remission can be achieved prior to transplantation. Similarly, DLI given for the treatment of post-transplant relapse is more successful when given during a second remission. In this study we report our results with a previously unreported approach consisting of short-term treatment with imatinib mesylate (formerly, STI571) to induce or maintain remission, followed by allogeneic transplantation or DLI and the impact on transplantation/DLI outcome. Sixteen patients were treated either in preparation for transplantation (n = 12), for DLI (n = 1), or for both (n = 3). Ten had CML in BC; seven myeloid and three lymphoid BC. Six patients had Ph(+) ALL. The donors were matched unrelated (n = 9), matched siblings (n = 5) or haplo-identical (n = 2). Eleven of 15 patients given imatinib pre-transplant were transplanted in complete hematologic response. Engraftment and GVHD rates were not different from expected. Seven patients had grade II-III hepatic toxicity after transplantation. After a median follow-up of 10 months (range, 3-16 months) six remain alive, two after further therapy. The 1 year survival rate was 25%. Four patients were given imatinib prior to DLI, all had complete response. Two remain in remission >6 months from relapse. In conclusion, treatment with imatinib allows transplantation in a more favorable status or maintaining remission with low toxicity until transplantation is feasible. Pre-transplant imatinib seems safe and not associated with excess post transplant complications. Imatinib may have substantial activity in combination with DLI. Further study of a larger group of patients is required to assess the impact on long-term outcome and the role of post-transplant imatinib in controlling residual disease. PMID- 12592326 TI - Effect of imatinib on haematopoietic recovery following idarubicin exposure. AB - SCF is a potent pro-proliferative cytokine crucial for haematopoiesis, which binds to c-kit and activates its tyrosine kinase activity. Inactivating mutations of either SCF or c-kit have been described in mice and lead to increased sensitivity to treatment with ionising radiation. Imatinib is a tyrosine kinase inhibitor with high affinity for c-Abl, PDGFR and c-kit. In this study we investigated the effect of concomitant administration of imatinib and idarubicin, an anthracycline with haematosuppressive activity, in nu/nu mice and murine bone marrow cells. Double-treated animals showed significantly increased mortality compared to mice that received imatinib or idarubicin alone only when idarubicin and imatinib were given simultaneously. The combined treatment induced a more severe neutropenia with a slower recovery when compared to mice treated with idarubicin alone. The myeloid metaplasia usually observed in the spleen after idarubicin treatment was absent in mice co-treated with imatinib. Bone marrow from double-treated animals also showed decreased numbers of megakaryocytes and myeloid precursor cells. In vitro culture of murine bone marrow cells in the presence of imatinib inhibited SCF-induced proliferation and recovery from treatment with idarubicin. Our results indicate that the simultaneous administration of imatinib enhances idarubicin-induced haematopoietic toxicity in vivo and in vitro. PMID- 12592327 TI - Application of different prognostic scoring systems and comparison of the FAB and WHO classifications in Korean patients with myelodysplastic syndrome. AB - We retrospectively studied 227 patients with MDS (1) to identify the prognostic factors of survival and acute leukemia evolution in Korean patients with MDS, (2) to apply different prognostic scoring systems to the same group of patients, and (3) to compare the FAB with the WHO classification. Six scoring systems were applied to the patients, and the FAB and WHO classifications were compared. The patients' median age was 57 years. The median survival time was 21 months, and age, dysgranulopoiesis and the IPSS cytogenetic groups were independent prognostic factors for survival. Acute leukemia occurred in 34 patients, and the cumulative incidence was 27.1% at 3 years. Marrow blast percentage was the only independent prognostic factor for acute leukemia evolution. Most scoring systems successfully discriminated risk groups for survival and acute leukemia evolution, but patient distribution into risk groups varied according to the scoring systems. Refractory cytopenia with multilineage dysplasia and RAEB II seemed to have different prognoses from RA or RARS and RAEB I, respectively. In summary, our MDS patients had different disease natures from those of Western countries regarding clinical features, prognostic factors and cytogenetic profiles. Although the WHO classification seems to improve the FAB classification, further studies are warranted to validate the utility of the WHO classification before it is accepted for routine clinical use. Our study has the limitations of retrospective analysis, and our results should be verified in future prospective studies. PMID- 12592328 TI - Treatment of relapsed or refractory acute myeloid leukemia with humanized anti CD33 monoclonal antibody HuM195. AB - HuM195 is a humanized, unconjugated, anti-CD33 monoclonal antibody. Fifty adult patients with relapsed or refractory AML were randomized to receive HuM195 at a dose of 12 or 36 mg/m(2) by intravenous infusion over 4 h on days 1-4 and 15-18. Patients with stable or responding disease received two additional cycles on days 29-32 and 43-46. HuM195 was given as first salvage therapy in 24 patients and as second or subsequent salvage therapy in 26 patients. Pretreatment blast percentage in the marrow was between 5 and 30% in 20 patients with the others having blast counts greater than 30%. The median age of patients was 62 years (range 26-86) and CD33 was detected in 95% of patients for whom immunophenotyping was available. Of 49 evaluable patients, two complete and one partial remission were observed. All three responses were in patients treated at the 12 mg/m(2) dose level and all had baseline blast percentages less than 30%. Decreases in blast counts ranging from 30 to 74% were seen in nine additional patients. Infusion-related events of fever and chills occurred in the majority of patients and were generally mild and primarily related to the first dose of antibody. No hepatic, renal or cardiac toxicities were observed and other adverse events such as nausea, vomiting, mucositis and diarrhea were uncommon or felt to be unrelated to HuM195. In addition, anti-HuM195 responses were not detected. HuM195 as a single agent has minimal, but observable, anti-leukemic activity in patients with relapsed or refractory AML and activity is confined to patients with low burden disease. No significant differences in clinical efficacy or toxicity were seen between the two dose levels of antibody. HuM195 was well tolerated with infusion related fevers and chills the predominant toxicities seen. Meaningful clinical efficacy of this unconjugated monoclonal antibody may be realized only in patients with minimal residual disease, or in combination with chemotherapy. PMID- 12592329 TI - Phase II trial of CPT-11 in myelodysplastic syndromes with excess of marrow blasts. AB - CPT-11 is an antineoplastic agent which acts as a specific inhibitor of DNA topisomerase 1 and has a broad spectrum of activity in solid tumors. Very few studies have evaluated the activity of CPT-11 in hematological malignancies. We conducted a phase II trial of CPT-11 in 26 patients with high-risk MDS (RAEB 1: n = 4; RAEB 2: n = 9; MDS having progressed to AML: n = 10; CMML: n = 3) who could not receive anthracycline/cytarabine intensive chemotherapy. Induction therapy consisted of four courses of CPT-11 given intravenously at 200 mg/m(2) every 2 weeks. Patient characteristics were: median age, 71 (range 51-77); sex, (M/F), 21/5, median % marrow blasts cells, 13.5 (range 7-52). Cytogenetics according to IPSS were: low-risk n = 13, intermediate-risk n = 6, high-risk n = 3, failure or not done n = 4. Six patients stopped treatment after only one or two courses of CPT-11 due to severe infection (n = 2), progressive disease (n = 3), acute lysis syndrome with renal failure (n = 1). In the 20 patients who received at least three cycles of CPT-11, complete remission was achieved in one case, partial remission in four cases, and hematological improvement in three cases with an overall response rate of 33% in the 26 patients. Duration of response was short (median 4 months, range 1-6 months) and median survival was 8 months (range 1-23 months). Digestive toxicity (diarrhea) occurred in 26/89 (29%) courses, but was mild (grade 1, 20% courses; grade 2 or 3, 9% courses). Hematological toxicity was difficult to assess in non-responders because of initial pancytopenia, but all the patients who responded had grade 3/4 hematological toxicity associated with grade >/=2 infection requiring hospitalization in 18% of the courses. No other major toxicity was observed. Thus CPT-11 has an interesting activity in MDS with excess of blasts; toxicity is easily managed and most patients can be treated in the out-clinic setting. These results suggest that further evaluation of CPT-11 in MDS is warranted. PMID- 12592330 TI - A phase II study of cladribine treatment for fludarabine refractory B cell chronic lymphocytic leukemia: results from CALGB Study 9211. AB - Cladribine has been reported to have little activity in fludarabine- refractory chronic lymphocytic leukemia (CLL). We sought to determine whether resistance to therapy with cladribine in fludarabine-refractory CLL patients represented primary drug resistance or the inability to tolerate the myelosuppression associated with this therapy. Patients with fludarabine refractory CLL patients without severe thrombocytopenia (platelets >/=50 x 10(9)/l) or granulocytopenia (neutrophils >1.5 x 10(9)/l) were enrolled. All patients received cladribine (0.14 mg/kg) as a 2-h intravenous infusion daily for 5 days, repeated every 4 weeks. Patients received up to six cycles of therapy. Twenty-eight patients enrolled; 13 had intermediate (Rai stage I or II) and 15 high (Rai stage III and IV) risk stages. No patient had a complete remission, but nine (32%; 95% confidence interval, 15-49%) attained a partial remission when assessed using the modified NCI criteria (1996). The median time to relapse for responders was 12 months, while median progression-free survival for the entire group was 9 months (95% confidence interval, 4-14 months). The median overall survival was 2.2 years (95% confidence interval, 0.8-3.1 years). Response was predicted by pre-treatment Rai status with seven of 13 (54%) intermediate risk vs two of 15 (13%) high-risk patients responding (P = 0.04). Toxicity was myelosuppression and infections (grade 3-5: neutropenia 75%, thrombocytopenia 68%, and infections 43%). Cladribine has modest clinical activity and considerable toxicity in a very selected group of patients with fludarabine-refractory CLL lacking pre-treatment neutropenia and thrombocytopenia. PMID- 12592331 TI - Benefit of high-dose methylprednisolone in comparison with conventional-dose prednisolone during remission induction therapy in childhood acute lymphoblastic leukemia for long-term follow-up. AB - Eight-year event-free survival (EFS) was evaluated in 205 patients with acute lymphoblastic leukemia (ALL), to consider the efficacy of high-dose methylprednisolone (HDMP) given during remission induction chemotherapy between 1 and 29 days. The St Jude Total XI Study protocol was used after some minor modifications in this trial. Patients were randomized into two groups. Group A (n = 108) received conventional dose (60 mg/m(2)/day orally) prednisolone and group B (n = 97) received HDMP (Prednol-L, 900-600 mg/m(2) orally) during remission induction chemotherapy. Complete remission was obtained in 95% of the 205 patients who were followed-up for 11 years; median follow-up was 72 months (range 60-129) and 8-year EFS rate was 60% overall (53% in group A, 66% in group B). The EFS rate of group B was significantly higher than of group A (P = 0.05). The 8 year EFS rate of groups A and B in the high-risk groups was 39% vs 63% (P = 0.002). When we compared 8-year EFS rate in groups A and B in the high-risk subgroup for both ages together /=10 years, it was 44% vs 74%, respectively. Among patients in the high-risk subgroup with a WBC count >/=50 x 10(9)/l, the 8-year EFS was 38% in group A vs58% in group B. During the 11-year follow-up period, a total of 64 relapses occurred in 205 patients. In group A relapses were higher (39%) than in group B (23%) (P = 0.05). These results suggest that HDMP during remission-induction chemotherapy improves the EFS rate significantly for high-risk patients in terms of the chances of cure. PMID- 12592332 TI - Treatment of SCID/human B cell precursor ALL with anti-CD19 and anti-CD22 immunotoxins. AB - The anti-CD19 (HD37-dgRTA) and anti-CD22 (RFB4-dgRTA) immunotoxins (ITs) are murine IgG(1) monoclonal antibodies (Mabs) conjugated to a deglycosylated ricin A chain (dgRTA). They are effective in killing B-lineage non-Hodgkin's lymphoma (NHL) cells in vitro, in vivo and in adult patients with B-lineage NHL. The potential of these agents for the treatment of childhood B-precursor acute lymphoblastic leukemia (ALL) is unknown. The anti-CD19 and anti-CD22 ITs should have anti-tumor activity against childhood B-lineage ALL since both target antigens are expressed on the surface of these cells. We have previously shown that, in vitro these two ITs selectively kill leukemia cells obtained from children with leukemia. To evaluate the efficacy of our ITs in an in vivo model we injected the human pre-B ALL cell line, NALM-6-UM1, into severe combined immunodeficient (SCID) mice. We tested the ability of two ITs to prolong survival or cure mice of both early and advanced tumors. In early disease, treatment with HD37-dgRTA, RFB4-dgRTA, or Combotox (an equimolar concentration of the two ITs) significantly improved their survival. In advanced disease, treatment with RFB4 dgRTA or Combotox significantly improved survival. Overall there were 10 long term survivors who were cured, as determined by survival beyond 150 days with no evidence of disease as determined by polymerase chain reaction (PCR) analysis. PMID- 12592333 TI - Early onset of chemotherapy can reduce the incidence of ATRA syndrome in newly diagnosed acute promyelocytic leukemia (APL) with low white blood cell counts: results from APL 93 trial. AB - Treatment combining ATRA and chemotherapy (CT) has improved the outcome of APL patients, by comparison with CT alone. ATRA syndrome is a life-threatening complication of ATRA treatment whose prophylaxis remains somewhat controversial. In APL93 trial, newly diagnosed APL patients CT) and ATRA with early addition of CT, on day 3 of ATRA treatment (ATRA + CT). The incidence of ATRA syndrome in the ATRA --> CT arm was 18% (22/122) as compared to 9.2% (17/184) in the ATRA + CT arm (P = 0.035). In the ATRA --> CT arm, three (2.5%) patients died from ATRA syndrome, as compared to one (0.5%) in the ATRA + CT group. Early addition of chemotherapy to ATRA in newly diagnosed APL with low WBC counts significantly reduced the incidence of ATRA syndrome. PMID- 12592334 TI - The emergence of a C/EBPalpha mutation in the clonal evolution of MDS towards secondary AML. AB - Recently, mutations in the transcription factor CCAAT/ enhancer binding protein alpha (C/EBPalpha) have been described in acute myeloid leukemia (AML). We performed a mutational analysis of the C/EBPalpha gene in the myelodysplastic syndromes and AML with antecedent MDS. No mutations were found in patients with refractory anemia (0/27), refractory anemia with ringed sideroblasts (0/7), refractory anemia with excess of blasts (RAEB 0/16) or chronic myelomonocytic leukemia (CMML 0/5). One out of 13 patients with RAEB-T/AML secondary to MDS showed a mutation in the C/EBPalpha gene. In this patient a 4 bp insertion disrupted codon 69 in one allele. This novel +1 frame shift is predicted to result in a truncated protein of 107 amino acids. However, the dominant protein translated was the C/EBPalpha isoform p30, which was previously shown to inhibit the DNA-binding and transactivation properties of C/EBPalpha p42. Interestingly this mutation could not be detected at diagnosis in the initial RAEB and RAEB-T stage. The mutation appeared at relapse after chemotherapy for RAEB-T. We conclude that the C/EBPalpha mutation was not essential for the initial blast accumulation. The emergence of a bast clone carrying a C/EBPalpha mutation at relapse indicates that this mutation may confer a growth advantage in a myeloid cell with an established differentiation block. PMID- 12592335 TI - Depsipeptide (FR 901228) promotes histone acetylation, gene transcription, apoptosis and its activity is enhanced by DNA methyltransferase inhibitors in AML1/ETO-positive leukemic cells. AB - In t(8;21) acute myeloid leukemia (AML), the AML1/ETO fusion protein promotes leukemogenesis by recruiting histone deacetylase (HDAC) and silencing AML1target genes important for hematopoietic differentiation. We hypothesized that depsipeptide (FR901228), a novel HDAC inhibitor evaluated in ongoing clinical trials, restores gene transcription and cell differentiation in AML1/ETO-positive cells. A dose-dependent increase in H3 and H4 histone acetylation was noted in depsipeptide-treated AML1/ETO-positive Kasumi-1 cells and blasts from a patient with t(8;21) AML. Consistent with this biological effect, we also showed a dose dependent increase in cytotoxicity, expression of IL-3, here used as read-out for silenced AML1-target genes, upregulation of CD11b with other morphologic changes suggestive of partial cell differentiation in Kasumi-1 cells. Some of these biologic effects were also attained in other myeloid leukemia cell lines, suggesting that depsipeptide has differentiation and cytotoxic activity in AML cells, regardless of the underlying genomic abnormality. Notably, the activity of depsipeptide was enhanced by 5-aza-2'-deoxycytidine, a DNA methyltransferase inhibitor (DNMT). These two agents in combination resulted in enhanced histone acetylation, IL-3 expression, and cytotoxicity, suggesting HDAC and DNMT activities as a potential dual target in future therapeutic strategies for AML1/ETO and other molecular subgroups of AML. PMID- 12592336 TI - Transcriptional activation is a key function encoded by MLL fusion partners. AB - Chromosomal translocations that fuse the mixed lineage leukemia gene (MLL) to a variety of unrelated partner genes are frequent in pediatric leukemias. The novel combination of genetic material leads to the production of active oncoproteins that depend on the contributions of both constituents. In a search for a common function amongst the diverse group of MLL fusion partners we constructed artificial fusions joining MLL with generic transactivator and repressor domains (acidic blob, GAL4 transactivator domain, Herpes simplex VP16 activation domain, KRAB repressor domain). Of all constructs tested, only MLL-VP16 was able to transform primary bone marrow cells and to induce a block of early myeloid differentiation like an authentic MLL fusion. Interestingly, the transformation capability of the artificial MLL fusions was correlated with the transcriptional potential of the resulting chimeric protein but it was not related to the strength of the isolated transactivation domain that was joined to MLL. These results prove for the first time that a general biological function - transactivation - might be the common denominator of many MLL fusion partners. PMID- 12592337 TI - The protein tyrosine phosphatase HePTP regulates nuclear translocation of ERK2 and can modulate megakaryocytic differentiation of K562 cells. AB - In response to PMA treatment K562 myelogenous leukemia cells undergo megakaryocytic differentiation, which is dependent on prolonged ERK activation and is characterized by growth arrest, upregulation of CD41 and IL-6, and, finally, by characteristic changes in cell morphology. The tyrosine phosphatase HePTP was recently demonstrated to regulate ERK activity and changes in HePTP expression have been associated with hematopoietic malignancies. Here, we have studied the function of HePTP during PMA-induced megakaryocytic differentiation of K562 cells. Overexpression of HePTP or inhibition of HePTP expression with antisense cDNA had no effect on PMA-induced cell cycle arrest or upregulation of cyclin D in K562 cells. The expression of megakaryocytic markers such as CD41 and IL6, however, were highly reduced in cells overexpressing HePTP, due to reduced ERK activation, and the cells were impaired in their ability to differentiate. Compared to control cells, HePTP antisense expressing cells did not show increased basal or PMA-induced ERK activity. However, antisense inhibition of HePTP enhanced nuclear translocation of ERK and the expression of the megakaryocytic markers CD41 and IL-6. Interestingly, like cells overexpressing HePTP, morphological differentiation was also impaired in HePTP antisense expressing cells. The results for the first time demonstrate that different aspects of megakaryocytic differentiation have distinct requirements for ERK activity. They further show that HePTP is involved in the regulation of nuclear translocation of ERK2 and that HePTP protein levels can modulate K562 cell differentiation. PMID- 12592338 TI - Constitutively active Akt1 protects HL60 leukemia cells from TRAIL-induced apoptosis through a mechanism involving NF-kappaB activation and cFLIP(L) up regulation. AB - TRAIL is a member of the tumor necrosis factor superfamily which induces apoptosis in cancer but not in normal cells. Akt1 promotes cell survival and blocks apoptosis. The scope of this paper was to investigate whether a HL60 human leukemia cell clone (named AR) with constitutively active Akt1 was resistant to TRAIL. We found that parental (PT) HL60 cells were very sensitive to a 6 h incubation in the presence of TRAIL and died by apoptosis. In contrast, AR cells were resistant to TRAIL concentrations as high as 2 microg/ml for 24 h. Two pharmacological inhibitors of PI3K, Ly294002 and wortmannin, restored TRAIL sensitivity of AR cells. AR cells stably overexpressing PTEN had lower Akt1 activity and were sensitive to TRAIL. Conversely, PT cells stably overexpressing a constitutive active form of Akt1 became TRAIL resistant. TRAIL activated caspase-8 but not caspase-9 or -10 in HL60 cells. We did not observe a protective effect of Bcl-X(L) or Bcl-2 against the cytotoxic activity of TRAIL, even though TRAIL induced cleavage of BID. There was a close correlation between TRAIL sensitivity and intranuclear presence of the p50 subunit of NF-kappaB. Higher levels of the FLICE inhibitory protein, cFLIP(L), were observed in TRAIL resistant cells. Both the cell permeable NF-kappaB inhibitor SN50 and cycloheximide lowered cFLIP(L)expression and restored sentivity of AR cells to TRAIL. Our results suggest that Akt1 may be an important regulator of TRAIL sensitivity in HL60 cells through the activation of NF-kappaB and up-regulation of cFLIP(L) synthesis. PMID- 12592339 TI - Differential activation of apoptosis regulatory pathways during monocytic vs granulocytic differentiation: a requirement for Bcl-X(L)and XIAP in the prolonged survival of monocytic cells. AB - Neutrophils and monocytes/macrophages are derived from common progenitors, but exhibit markedly different lifespans. Differentiated neutrophils are short-lived and die rapidly by apoptosis, while monocytic cells are longer-lived. In this report we used the HL-60 cell line as a model system to identify differences in apoptotic pathways which might account for the differing lifespans of granulocytic vs monocytic cells. We observed that induction of granulocytic differentiation by retinoic acid led to robust activation of the executioner protease caspase-3, and early onset of apoptosis. By contrast, caspase-3 was not appreciably activated during phorbol 12-myristate 13-acetate (PMA)-induced monocytic differentiation, and apoptosis was delayed in these cells. Since the activation of caspase-3 is inhibited by members of the inhibitor of apoptosis (IAP) and Bcl-2 protein families, we investigated the expression of anti apoptotic members of these families. Induction of monocytic differentiation led to marked upregulation of the IAP protein XIAP, as well as the Bcl-2 family member Bcl-X(L). During granulocytic differentiation the levels of XIAP progressively declined, while Bcl-X(L) levels remained unchanged. A different IAP protein, survivin, was downregulated during differentiation along either lineage, as was expression of Bcl-2. The upregulation of Bcl-X(L) during monocytic differentiation coincided with phosphorylation/activation of STAT3, a known activator of bcl-X gene transcription. Moreover, Bcl-X(L) upregulation was dependent on MEK/ERK signaling. Upregulation of XIAP proceeded in a MEK/ERK independent fashion. Treatment with antisense Bcl-X(L) or XIAP oligonucleotides resulted in significant loss of viability in cells differentiating along the monocytic lineage. Together, these findings indicate that the levels of XIAP and Bcl-X(L) are regulated by distinct pathways during monocytic differentiation, and that upregulation of these proteins contributes to the increased longevity of cells in the monocytic lineage. PMID- 12592340 TI - Telomere dynamics in childhood leukemia and solid tumors: a follow-up study. AB - Telomeres of hematopoietic cells shorten with age, possibly contributing to the aging-associated hematopoietic pathology (immunosenescence, malignant transformation). Accelerated telomere shortening is seen with replicative stress, such as during administration of serial chemotherapy cycles for the treatment of childhood cancer. To define the long-term consequences of pediatric cancer treatment on hematopoietic cell telomere length, we undertook a prospective 4 year follow-up study of a 61-patient cohort of pediatric malignancies in a community-based setting. We found that mononuclear cells (MNC) and granulocytes of children with standard-risk acute lymphoblastic leukemia (ALL) suffered minimal telomere shortening throughout therapy (less than 1 kbp; average follow up, 20 months), while those of children with solid tumors showed greater and more heterogenous telomere attrition (0.5-2.8 kbp, average follow-up, 9 months). In addition, we evaluated the role of telomerase, the enzyme commonly up-regulated in pediatric leukemic and solid tumor cells for telomere length maintenance, as a disease marker. Serial determinations of telomerase in MNC were useful to confirm disease remission in leukemia, but play no role in the follow-up of children with solid tumors. PMID- 12592341 TI - Microsatellite instability and hMLH1 promoter hypermethylation in Richter's transformation of chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia (CLL) is an indolent B cell non-Hodgkin lymphoma (NHL) that may transform into diffuse large B cell lymphoma (DLBL). This transformation is referred to as Richter's syndrome or transformation. To analyze whether microsatellite instability (MSI) and DNA mismatch repair defects are associated with Richter's transformation, we have performed microsatellite analysis, mutational analysis of hMLH1 and hMSH2 genes and methylation status analysis of CpG island of the hMLH1 promoter on serial biopsy specimens from 19 patients with CLL. Ten cases of CLL showed no histologic alteration in the second biopsy, and nine cases of CLL underwent morphologic transformation to DLBL in the second biopsy. Using eight microsatellite loci, high level of MSI was associated with Richter's transformation in four cases of CLL, but none of the CLLs displayed this level of MSI without transformation. Mutations of the hMLH1 or hMSH2 genes were not detected in any of the lymphoma samples. In five cases of Richter's transformation the hMLH1 promoter was hypermethylated in both CLL and DLBL samples. Hypermethylation of the hMLH1 promoter associated with high-level of MSI in four cases, and low-level of MSI in one case. These results suggest that in certain cases of Richter's transformation the DNA mismatch-repair defect initiated genetic instability may play a role in tumor progression. PMID- 12592342 TI - False leukemia-lymphoma cell lines: an update on over 500 cell lines. AB - Human leukemia-lymphoma (LL) cell lines represent an extremely important resource for research in a variety of fields and disciplines. As the cell lines are used as in vitro model systems in lieu of primary cell material, it is crucial that the cells in the culture flasks faithfully correspond to the purported objects of study. Obviously, proper authentication of cell line derivation and precise characterization are indispensable requirements to use as model systems. A number of studies has shown an unacceptable level of LL cell lines to be false. We present here the results of authenticating a comprehensively large sample (n = 550) of LL cell lines mainly by DNA fingerprinting and cytogenetic evaluation. Surprisingly, near-identical incidences (ca 15%) of false cell lines were observed among cell lines obtained directly from original investigators (59/395: 14.9%) and from secondary sources (23/155: 14.8%) implying that most cross contamination is perpetrated by originators, presumably during establishment. By comparing our data with those published, we were further able to subclassify the false cell lines as (1) virtual: cross-contaminated with and unretrievably overgrown by other cell lines during initiation, never enjoying independent existence; (2) misidentified: cross-contaminated subsequent to establishment so that an original prototype may still exist; or (3) misclassified: unwittingly established from an unintended (often normal) cell type. Prolific classic leukemia cell lines were found to account for the majority of cross contaminations, eg CCRF-CEM, HL-60, JURKAT, K-562 and U-937. We discuss the impact of cross-contaminations on scientific research, the reluctance of scientists to address the problem, and consider possible solutions. These findings provide a rationale for mandating the procurement of reputably sourced LL cell lines and their regular authentication thereafter. PMID- 12592343 TI - Chromosome abnormalities clustering and its implications for pathogenesis and prognosis in myeloma. AB - The nonrandom recurrent nature of chromosome abnormalities in myeloma suggests a role for them in disease pathogenesis. We performed a careful cytogenetic analysis of patients with abnormal karyotypes (n = 254), to discern patterns of association, search for novel abnormalities and elucidate clinical implications. Patients with karyotypic abnormalities suggestive of myelodysplasia/acute leukemia were excluded. In this study we compared survival by abnormality only between patients with abnormal karyotypes. Patients with abnormalities were more likely to have features of aggressive disease as compared to all other patients without abnormalities entered into the myeloma database (lower hemoglobin, higher beta(2)-microglobulin, labeling-index and plasmocytosis; all P < 0.0001). Several groups of patients could be readily identified; hypodiploid (22%), pseudodiploid (36%), hyperdiploid (31%) and near-tetraploid (11%). Clustering associations were seen among several trisomies and monosomy of chromosome 13 and 14. Several monosomies (-2, -3, -13, -14 and -19), 1p translocations/ deletions, and hypodiploidy were associated with a significantly shorter survival. Trisomy of chromosome 13 was rare ( <2%). Even among patients with abnormal karyotypes, specific chromosome abnormalities can impart biologic variability in myeloma, including several monosomies, hypodiploidy and abnormalities of 1p. PMID- 12592344 TI - Mobilization of peripheral blood progenitor cells with a combination of cyclophosphamide, r-metHuSCF and filgrastim in patients with breast cancer previously treated with chemotherapy. AB - The objective of our study was to determine the effect of adding r-metHuSCF to Filgrastim and cyclophosphamide for mobilization of peripheral blood progenitor cells (PBPC), on collection of CD34(+) cells and engraftment after autologous stem cell transplant. Twenty-three patients with previously treated stage II-IV breast cancer received cyclophosphamide (3 g/m(2)), Filgrastim 5 microg/kg daily and r-metHuSCF 20 microg/kg daily. Two PBPC collections were performed on consecutive days starting the day the WBC count was above 7.5 x 10(3)/microl. Collection was performed between days +9 and +12 and the median number of CD34(+) cells collected was 9.9 x 10(6)/kg (1.1-53.1) and 6.6 x 10(6)/kg (1.4-33.8) for the first and second apheresis, respectively. Despite being previously treated patients, the target CD34(+) cell dose required for SCT was obtained in all patients. SCT was associated with rapid neutrophil and platelet engraftment and a highly significant correlation was observed between the number of CD34(+) cells infused and engraftment. Treatment with SCF plus filgrastim was well tolerated, with mild to moderate local skin rash being the most frequently reported adverse event. In conclusion, addition of r-metHuSCF induces mobilization of a large number of CD34(+) cells which results in shortening of time to engraftment and hospitalization. PMID- 12592345 TI - IL-4 and interferon gamma regulate expression of inducible nitric oxide synthase in chronic lymphocytic leukemia cells. AB - Chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of long lived non-dividing CD5(+) B cells. Nitric oxide (NO) is an important regulator of apoptosis, and the viability of cultured B-CLL cells may be dependent on the autocrine production of nitric oxide by inducible nitric oxide synthase (NOS2). We performed this study to determine whether cytokine factors that prevent spontaneous in vitroapoptosis of B-CLL cells induce B-CLL cell NOS2 enzyme activity. B-CLL cells expressed NOS enzyme activity and NOS2 protein and mRNA. IL 4 and IFN-gamma increased B-CLL cell NOS2 enzyme activity and protein expression during in vitro culture. IFN-gamma, but not IL-4, increased NOS2 mRNA expression in cultured B-CLL cells suggesting that IL-4-mediated changes of NOS2 protein expression occurred at the post-transcriptional level. We were unable to detect increased concentrations of nitrite or nitrate (NO(x)) as surrogate markers of NO production in B-CLL cell cultures treated with IL-4 or IFN-gamma. IL-4 and IFN gamma diminished NOS inhibitor-induced B-CLL cell death. In summary, we found that B-CLL cells expressed NOS2 and that IL-4 and IFN-gamma increased B-CLL NOS2 expression. Cytokine-mediated expression of NOS2 by B-CLL cells may promote their survival, and therapeutic strategies that target NOS2 or quench NO may be beneficial in patients with B-CLL. PMID- 12592346 TI - The farnesyl transferase inhibitor, FTI-277, inhibits growth and induces apoptosis in drug-resistant myeloma tumor cells. AB - Mutations of the ras gene are among the most commonly identified transforming events in human cancers, including multiple myeloma. Farnesyltransferase inhibitors (FTI) were developed to prevent Ras processing and induce cancer cell death. Several FTIs are in phase II and one is in phase III clinical trials. Preclinically, most of the focus has been on solid tumors, and the effects of FTIs in multiple myeloma have not been investigated. In this study we examined the cytotoxic activity and inhibition of Ras processing in three myeloma cell lines with differing Ras mutation status. H929 cells with activated N-Ras were more sensitive to FTI-277 treatment than 8226 and U266 cells with activated K-Ras or wild-type Ras, respectively. A combination of FTI-277 and a geranylgeranyltransferase I inhibitor (GGTI)-2166 inhibited K-Ras processing and enhanced cell death in 8226 cells. U266 cells and Bcl-x(L) transfectants were equally sensitive to FTI-277 treatment. Similarly, 8226 cells selected for resistance to various chemotherapeutic agents, which resulted in either P glycoprotein overexpression, altered topoisomerase II activity, or elevated glutathione levels, were equally sensitive to FTI-277. These preclinical studies suggest that prenylation inhibitors may represent new therapeutic agents for the treatment of refractory or drug-resistant multiple myeloma. PMID- 12592347 TI - Imatinib mesylate induces durable complete remission of advanced CML persisting after allogeneic bone marrow transplantation. PMID- 12592348 TI - Occurrence of additional chromosome aberrations in chronic myeloid leukemia patients treated with imatinib mesylate. PMID- 12592350 TI - Sustained, clonal karyotype abnormalities in the Philadelphia chromosome negative cells of CML patients successfully treated with Imatinib. PMID- 12592349 TI - Imatinib mesylate-induced molecular remission of Philadelphia chromosome-positive myelodysplastic syndrome. PMID- 12592352 TI - Contrary to adult patients, expression of the multidrug resistance gene (MDR1) fails to define a poor prognostic group in childhood AML. PMID- 12592351 TI - Gemtuzumab ozogamicin in pediatric CD33-positive acute lymphoblastic leukemia: first clinical experiences and relation with cellular sensitivity to single agent calicheamicin. PMID- 12592353 TI - C-Kit point mutations in core binding factor leukemias: correlation with white blood cell count and the white blood cell index. PMID- 12592354 TI - Deletion of the 5' abl region in Philadelphia chromosome-positive chronic myeloid leukemia. PMID- 12592355 TI - Engraftment of allogeneic mesenchymal stem cells in the bone marrow of a patient with severe idiopathic aplastic anemia improves stroma. PMID- 12592356 TI - Frequency of ETV6/AML1 fusion in adult acute lymphoblastic leukemia. PMID- 12592359 TI - Apoptosis, ageing and cancer susceptibility. AB - We have previously shown that peripheral blood lymphocytes (PBL) from individuals carrying a germline TP53 mutation show a dramatically reduced apoptotic response to radiation. As part of a study of this phenomenon, we also investigated apoptotic response in a series of breast cancer patients lacking TP53 mutations and in a control group of individuals without cancer. There was a significant reduction in mean apoptotic response with increasing age in all groups. These findings are consistent with a number of studies in rodents, which have demonstrated a reduction in DNA damage-induced apoptosis with increasing age. In addition, after adjusting for age, breast cancer patients showed significantly reduced apoptotic responses compared with normal controls (P=0.002). The odds ratio for breast cancer in women with an apoptotic response of <35%, compared with women with a response of >49%, was 6.42 (95% CI 1.68-24.6). The data further support the hypothesis that a reduction in apoptotic response to DNA damage with increasing age may play a significant role in the age-related increase in cancer. PMID- 12592360 TI - Phase I/II study of gemcitabine plus mitoxantrone as salvage chemotherapy in metastatic breast cancer. AB - The purpose of this study was to determine the maximum-tolerated dose of gemcitabine plus mitoxantrone in women with metastatic breast cancer (MBC) and to evaluate activity and toxicity of this combination in a phase II trial. Sixty three patients with MBC, previously treated with chemotherapy including anthracycline and/or taxanes, were treated with mitoxantrone 10 or 12 mg m(-2) intravenously on day 1 plus gemcitabine in escalating doses from 600 to 1200 mg m(-2) intravenously on days 1 and 8, every 3 weeks. In phase I, on 23 patients entered on study, dose-limiting toxicity occurred at the dosage of 1200 mg m(-2) gemcitabine and 10 mg m(-2) mitoxantrone, with three out of five patients developing grade 4 neutropenia. In phase II, with gemcitabine administered at 1000 mg m(-2) and mitoxantrone at 10 mg m(-2), 12 (30%) out of 40 assessable patients responded, even if no complete response was obtained. Moreover, stable disease was observed in eight (20%) patients. The median time to treatment failure was 22 weeks (range, 2-33), and median survival was 42 weeks (range, 2 92). Grade 3 and 4 neutropenia were observed in 12 (30%) and one (2.5%) cases respectively; grade 3 thrombocytopenia was observed in two patients (5%), grade 2 mucositis in two patients (5%), grade 3 anaemia in two patients (5%), grade 3 alopecia in one patient (2.5%) and asymptomatic cardiotoxicity in three patients (8%), respectively. In conclusion, the doses of 10 mg m(-2) (day 1) for mitoxantrone and 1000 mg m(-2) for gemcitabine (days 1-8) every 3 weeks resulted active and safe in MBC. Further investigations in less heavily pretreated patients are warranted. PMID- 12592362 TI - Impact of medical specialists' locus of control on communication skills in oncological interviews. AB - Although is it widely recognised that physicians' characteristics could influence their communication styles, no empirical evidence is currently available. No studies are available on the impact of physicians' locus of control (LOC) on their communication skills. LOC is a generalised belief regarding the extent to which life outcomes are controlled by an individual's actions (internal LOC) or by external forces such as luck, fate or other individuals (external LOC). It was hypothesised that physicians with external LOC would take more into account others' concerns than physicians with internal LOC and would consequently use more appropriate assessment, informative and supportive functions. A total of 81 medical specialists were assessed in a simulated interview and a clinical interview. Communication skills were rated according to the Cancer Research Campaign Workshop Evaluation Manual. LOC was assessed using the Rotter I-E scale. Communication skills of the upper and lower quartiles of physicians in respect of their scores on this scale were compared using Student's t-test. Results show that physicians with external LOC give more appropriate information than physicians with internal LOC in simulated interviews (P=0.011) and less premature information than physicians with internal LOC in clinical interviews (P=0.015). This result provides evidence that physicians' LOC can influence their communication styles in oncological interviews and in particular the way they provide information to the patient. PMID- 12592361 TI - Survival with dacarbazine and fotemustine in newly diagnosed glioblastoma multiforme. AB - A total of 55 patients with histologically proven glioblastoma multiforme (total gross resection: n=24, subtotal resection: n=20, stereotactic biopsy: n=11) were treated with the combination of dacarbazine (D) (200 mg m(-2)) and fotemustine (F) (100 mg m(-2)) and concomitant radiotherapy (2 Gy day(-1), 5 days per week using limited fields up to 60 Gy) to assess efficacy and toxicity of this regimen. Survival (median survival, 12-, 18- and 24-month survival rates) and time to progression (median time to progression (TTP), 6-month progression-free survival) were analysed by Kaplan-Meier's method. A total of 268 (range 1-8, median: 5) cycles were administered. Median survival is 14.5+ (range: 0.5-40+) months, and the 12-, 18- and 24-month survival rates are 58, 29 and 23%, respectively. Median TTP from the start of D/F therapy is 9.5+ (range: 0.5-33+) months. The 6-month progression-free survival is 54%. Partial remissions were observed in 3.6%. Main toxicity was thrombocytopenia. Five patients were excluded from further D/F application, four patients because of prolonged thrombocytopenia NCI-CTC grades 3 and 4 and one patient because of whole body erythrodermia. One patient died because of septic fever during thrombocytopenia and leukopenia NCI CTC grade 4 after the first cycle. No other toxicities of NCI-CTC grade 3 or 4 occurred. The treatment is feasible in a complete outpatient setting and the results of the D/F regimen justify further investigations with these compounds. PMID- 12592363 TI - Molecular classification of synovial sarcomas, leiomyosarcomas and malignant fibrous histiocytomas by gene expression profiling. AB - In this study, we have used genome-wide expression profiling to categorise synovial sarcomas, leiomyosarcomas and malignant fibrous histiocytomas (MFHs). Following hierarchical clustering analysis of the expression data, the best match between tumour clusters and conventional diagnosis was observed for synovial sarcomas. Eight of nine synovial sarcomas examined formed a cluster that was characterised by higher expression of a set of 48 genes. In contrast, sarcomas conventionally classified as leiomyosarcomas and MFHs did not match the clusters defined by hierarchical clustering analysis. One major cluster contained a mixture of both leiomyosarcomas and MFHs and was defined by the lower expression of a set of 202 genes. A cluster containing a subgroup of MFHs was also detected. These results may have implications for the classification of soft tissue sarcomas, and are consistent with the view that sarcomas conventionally defined as MFHs do not represent a separate diagnostic category. PMID- 12592364 TI - Clinical implications of quantitative real-time RT-PCR analysis of hTERT gene expression in human gliomas. AB - The presence of telomerase activity in a glioma may be a predictor of its malignant potential. Activation of telomerase is regulated at the transcriptional level of the human telomerase reverse transcriptase (hTERT). Here, we evaluated whether the amount of hTERT mRNA provides a molecular marker of glioma malignancy that would have clinical utility. We used a real-time RT-PCR to assess the number of hTERT transcripts in primary tumour samples derived from 70 glioma patients. Results were standardised by quantifying the number of ABL transcripts as internal control and expressed as hTERT/ABL ratio. The percentage of patients with detectable hTERT mRNA markedly increased with enhanced malignancy: low-grade gliomas expressed hTERT in one out of 14 cases (7.1%), anaplastic gliomas in four out of 13 cases (30.8%) and glioblastoma multiforme (GBM) tumours in 30 out of 43 cases (69.8%). The mean hTERT/ABL ratio was significantly higher in GBMs than in non-GBMs. Subdividing hTERT/ABL ratios as low (< pr = 25%) and high (>25%), we found that the overall survival among hTERT-positive GBMs was significantly worse in high hTERT expressors than in low hTERT expressors (P=0.0082). We conclude that the amount of hTERT mRNA may represent a diagnostic and prognostic indicator for GBM patients. PMID- 12592365 TI - CpG methylation of MGMT and hMLH1 promoter in hepatocellular carcinoma associated with hepatitis viral infection. AB - Inactivations of DNA repair genes, O(6)-methylguanine-DNA methyltransferase (MGMT) and hMLH1, by promoter hypermethylation have been reported in several types of primary human neoplasia. This epigenetic inactivation mechanism remains elusive in hepatocellular carcinoma (HCC). To investigate the relation between the expression of MGMT and hMLH1 and the CpG methylation within their promoters in HCCs with or without hepatitis viral infection, we performed immunohistochemistry and urea/bisulphite sequencing on 46 HCCs, corresponding noncancerous tissues, and 20 normal liver tissues. MGMT- and hMLH1-negative HCCs were 60.9% (28 out of 46) and 21.8% (10 out of 46), respectively. HCCs lacking both proteins were 10.9% (five out of 46). The frequency and extent of CpG methylation in the MGMT promoter increased along with hepatitis viral infection and pathological progression. MGMT-negative tumours showed very frequent and widespread methylation in the promoter compared with MGMT-positive tumours. Half of the hMLH1-negative HCCs showed promoter hypermethylation. These data suggested that MGMT gene silencing in a subset of HCCs was likely caused by epigenetic alteration, such as promoter hypermethylation, and that the promoter hypermethylation silenced the hMLH1 gene in half of the hMLH1-negative tumours. A correlation between the promoter methylation status and viral infection, although it was weak, intimated that hepatitis viral infections could play a role in the CpG methylation of the MGMT promoter. PMID- 12592366 TI - Specific T-cell immunity against Ki-ras peptides in patients with pancreatic and colorectal cancers. AB - Mutations of codon 12 in the Ki-ras gene are frequently found in pancreatic and colorectal cancers. It has been demonstrated that human T-cells have the potential to recognise tumours expressing mutated ras-derived peptides. However, it remains unclear whether T-cells from a given individual can recognise the mutant peptides, which are expressed in that individual's tumour tissues. Mutations of the Ki-ras oncogene were analysed by the mutant-allele-specific amplification (MASA) method in pancreatic and colorectal tumour tissues, and T cell responses against mutated Ki-ras-derived peptides were measured by [(3)H]thymidine incorporation and IFN-gamma production assays. Specific T-cell responses against Ki-ras-products were found in cancer patients, whereas no immune response was observed in normal individuals (P<0.01). Six of the eight pancreatic cancer patients (75%) and nine of 26 colorectal cancer patients (35%) had T-cell responses to mutated Ki-ras-derived-peptides. T-cell response in a given individual cannot recognise the same mutated ras peptide, which is expressed in that individual's tumour tissues. However, pancreatic and colorectal cancer patients have T-cell immunity against Ki-ras-peptides, and this provides potential target for cancer immunotherapy. PMID- 12592367 TI - Independent prognostic value of fascin immunoreactivity in stage I nonsmall cell lung cancer. AB - Fascin-1, the most expressed form of fascin in vertebrate tissues, is an actin bundling protein that induces cell membrane protrusions and increases motility of normal and transformed epithelial cells. Very few data are available on the role of this protein in nonsmall cell lung cancer (NSCLC). Two hundred and twenty patients with stage I NSCLC and long-term follow-up were evaluated immunocytochemically for fascin expression. Overall, variable fascin immunoreactivity was detected in 98% of 116 squamous cell carcinomas, in 78% of 96 adenocarcinomas, in 83% of six large cell carcinomas, and in the two adenosquamous carcinomas under study. Neoplastic emboli were commonly decorated by the antifascin antibody (P<0.001), also when the surrounding invasive carcinoma was unreactive. Fascin immunoreactivity correlated with high tumour grade (P=0.017) and, in adenocarcinomas, with high Ki-67 labelling index (P=0.021). Adenocarcinomas with a prevalent bronchiolo-alveolar in situ component were less commonly immunoreactive for fascin than invasive tumours (P=0.005). Contralateral thoracic or distant metastases were associated significantly with diffuse (>60% immunoreactive tumour cells) fascin expression in adenocarcinomas (P=0.043), and marginally with strong fascin immunostaining in squamous cell carcinomas (P=0.13). No associations were noted with any other clinicopathological variables tested. Patients with tumours showing diffuse (>60% immunoreactive neoplastic cells) and/or strong immunoreactivity for fascin had a shorter survival (P=0.006 for adenocarcinomas and P=0.026 for squamous cell carcinomas), even after multivariate analysis (P=0.014 and 0.050, respectively). The current study documents for the first time that fascin is upregulated in invasive and more aggressive NSCLC, being an independent prognostic predictor of unfavourable clinical course of the disease. Targetting the fascin pathway could be a novel therapeutic strategy of NSCLC. PMID- 12592368 TI - Particulate debris from a titanium metal prosthesis induces genomic instability in primary human fibroblast cells. AB - Previous studies detected both lethal and cumulative chromosomal aberrations in bone marrow and peripheral blood of patients with worn hip and knee replacements. This study shows that wear debris from a worn titanium metal on high-density polyethylene hip replacement also produces chromosomal instability and reproductive failure in cell culture. The progeny of these treated cells also displayed chromosomal instability, mainly consisting of chromatid breaks and minutes, and reproductive failure as determined by clonogenic survival many generations postexposure. These delayed effects are similar to those caused by the heavy metals cadmium and nickel and to those seen for low-dose radiation. These findings may have important implications with regard to the long-term risks of joint replacement surgery. This highlights the need for long-term epidemiological studies of patients with surgical implants. PMID- 12592369 TI - Vascular patterns in reactive lymphoid tissue and in non-Hodgkin's lymphoma. AB - The few studies published on angiogenesis in lymphoma have raised the question of whether or not microvessel density (MVD) is associated with more aggressive disease and have reported the observation that in follicular lymphomas, vessels are mature rather than immature. We investigated MVD and the vascular phenotype within follicular or diffuse large B-cell lymphomas, reactive nodes and tonsils. Vascular phenotype was defined by the expression or loss of reactivity to the antibody LH39 (detecting the LH39 laminin epitope of the basement membrane in mature vessels) and by detection of alpha V beta 3 (expressed on immature vessels). In reactive nodes and in follicular lymphomas, MVD was higher in the paracortex than in germinal centres or in neoplastic follicles. However, in neoplastic follicles an increase in alpha V beta 3-positive endothelium suggested the activation of an angiogenic pathway different from that present in the reactive follicles. In large B-cell lymphomas, MVD was higher than in reactive and neoplastic follicles but lower than in the reactive paracortex. The number of immature vessels (LH39 negative) and of alpha V beta 3-positive vessels was higher than in reactive lymph nodes and follicular lymphoma suggesting that a switch to a different angiogenic pathway has occurred. Finally, we have demonstrated that within reactive and neoplastic follicles vascular regression is occurring, perhaps constraining the growth of reactive follicles alongside other phenomena such as apoptosis. Vascular regression was previously believed to occur in adults only in ovarian and endometrial tissue. We conclude that different types of angiogenesis are present in follicular lymphomas and large B-cell lymphomas. This has implications for possible future therapies. PMID- 12592370 TI - Thin-layer liquid-based cervical cytology and PCR for detecting and typing human papillomavirus DNA in Flemish women. AB - The objective of this study was to document the occurrence and to correlate the prevalence of different human papillomavirus (HPV) types with the cytological results on simultaneously performed thin-layer preparations in a large population of Flemish women. During 1 year, 69 290 thin-layer preparations were interpreted using the Bethesda classification system. Using an algorithm for HPV testing based on consensus primers and type-specific PCRs in combination with liquid based cytology, we determined the occurrence and distribution of 14 different oncogenic HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68). Reflex HPV testing was performed on cytologically abnormal samples and on an age matched randomly selected control group with normal cervical cytology (n=1351). Correlation between cytology, age and prevalence for the 14 different high-risk HPV types is given. There is a significant increase in predominance of high-risk HPV types, with increasing abnormal cytology. Coinfection with multiple HPV types also increased with cytological abnormalities, and was highest in HSIL (16.7%). In Flanders, HSIL was most often associated with HPV types 16, 33, 35, 31, 18 and 51. Using thin-layer liquid-based cytology and PCR to detect HPV, it is feasible to screen large numbers of women. PMID- 12592371 TI - The parathyroid hormone-related protein receptor is expressed in breast cancer bone metastases and promotes autocrine proliferation in breast carcinoma cells. AB - Overproduction of parathyroid hormone-related protein (PTHRP) occurs in a high proportion of primary breast cancers (PBC) and is strongly implicated in their metastatic spread to bone. Although the PTHRP-receptor (PTHRP-R) is often coexpressed with PTHRP in PBC, its role in regulating breast cancer cell proliferation and metastases to bone remains unclear. The aims of this study were to determine the expression of the PTHRP-R in breast cancer bone metastases (BM) and to investigate the effects of PTHRP-R overexpression on breast cancer cell proliferation. PTHRP-R expression occurred in 85% (11 out of 13) of BM compared with 58% (39 out of 67) of PBC. Median expression was higher (P<0.05) in BM compared with PBC. PTHRP increased cAMP accumulation and DNA synthesis in MCF-7 cells stably overexpressing the PTHRP-R (MCF-7(WTR)) but not in MCF-7(VEC) control cells. The increase in DNA synthesis was mimicked by the cAMP pathway activator forskolin. The receptor antagonist PTHRP(7-34) reduced DNA synthesis in MCF-7(WTR) cells, but not MCF-7(VEC) cells, indicating that receptor overexpression promotes autocrine PTHRP activity. MCF-7(WTR) cells showed increased mitogenic responsiveness to fetal calf serum and reduced doubling times. PTHRP induced weak activation of ERK1 and ERK2 and potentiated their activation by serum growth factors. Collectively these results show that the PTHRP-R is frequently expressed in breast cancer BM and indicate that receptor overexpression drives proliferation via autocrine signals that are mediated via cAMP and ERK pathways. PMID- 12592372 TI - Correlation of COX-2 and Ep-CAM overexpression in human invasive breast cancer and its impact on survival. AB - Recent studies have demonstrated cyclooxygenase 2 (COX-2) overexpression in various human malignancies, especially in breast cancer, where COX-2 turned out to be a predictor of poor survival. To evaluate the relation of COX-2 and Ep-CAM overexpression and its prognostic significance, we performed a retrospective study on 212 breast cancer patients with a median follow-up time of 10.5 years. Overexpression of COX-2 in tumour tissue samples was assessed by immunohistochemistry. COX-2 overexpression was found in 48.6% of the tumour samples and was predictive for poor disease-free and overall survival. Univariate analysis revealed a strong correlation between COX-2 and Ep-CAM overexpression (P=0.009). Concurrent COX-2 and Ep-CAM overexpression was present in 21.7% of tumour specimens and had an additive negative impact on disease-free and overall survival. Determination of both tumour markers should help in guiding new therapeutic strategies in patients with invasive breast cancer. PMID- 12592373 TI - hAG-2 and hAG-3, human homologues of genes involved in differentiation, are associated with oestrogen receptor-positive breast tumours and interact with metastasis gene C4.4a and dystroglycan. AB - hAG-2 and hAG-3 are recently discovered human homologues of the secreted Xenopus laevis proteins XAG-1/2 (AGR-1/2) that are expressed in the cement gland, an ectodermal organ in the head associated with anteroposterior fate determination during early development. Although the roles of hAG-2 and hAG-3 in mammalian cells are unknown, both proteins share a high degree of protein sequence homology and lie adjacent to one another on chromosome 7p21. hAG-2 mRNA expression has previously been demonstrated in oestrogen receptor (ER)-positive cell lines. In this study, we have used real-time quantitative RT - PCR analysis and immunohistochemistry on tissue microarrays to demonstrate concordant expression of hAG-2 and hAG-3 mRNA and protein in breast tumour tissues. Tumour expression of both genes correlated with OR (hAG2, P=0.0002; hAG-3, P=0.0012), and inversely correlated with epidermal growth factor receptor (EGFR) (P=0.003). Yeast two hybrid cloning identified metastasis-associated GPI-anchored C4.4a protein and extracellular alpha-dystroglycan (DAG-1) as binding partners for both hAG-2 and hAG-3, which if replicated in clinical oncology would demonstrate a potential role in tumour metastasis through the regulation of receptor adhesion and functioning. hAG-2 and hAG-3 may therefore serve as useful molecular markers and/or potential therapeutic targets for hormone-responsive breast tumours. PMID- 12592374 TI - Proapoptotic genes BAX and CD40L are predictors of survival in transitional cell carcinoma of the bladder. AB - The purpose of the study was to investigate the effects of expression of a range of genes involved in apoptosis on outcome in bladder cancer. Immunohistochemistry was used to examine expression of BCL2, BAX, P53, CD40 and CD40L in archival tissues of patients included in various treatment trials for transitional cell carcinoma (TCC) of the bladder. Data were collected on 94 patients who first presented with either invasive or superficial bladder cancer. Median follow-up for alive patients was 83 months (m) (range 12-195 m). Median survival was 80 m (95% CI=56-128 m). Median survivals for the various markers were as follows: BAX positive patients 110 m vs BAX-negative patients 18 m (P=0.0002); CD40L-positive patients 95 m vs CD40L-negative patients 45 m (P=0.04); BCL2-positive patients 44 m and BCL2-negative patients 74 m, (P=0.64); CD40-positive patients 110 m and CD40 negative patients 45 m (P=0.12); and P53 positive patients 80 m and P53 negative patients 45 m (P=0.58). In conclusion, it was seen that overexpressions of BAX and CD40L are prognostic of better survival in TCC of the bladder. Our results also raise the possibility of the future development of CD40- and CD40 ligand-based immunotherapy for bladder cancer. This study links proapoptotic and antiapoptotic markers to overall survival. PMID- 12592375 TI - Association between telomere length and V(H) gene mutation status in chronic lymphocytic leukaemia: clinical and biological implications. AB - The immunoglobulin V(H) gene mutation status can divide B-cell chronic lymphocytic leukaemia (CLL) into two entities with a different clinical course. Cases with unmutated V(H) genes, considered to evolve from pregerminal centre (GC) cells, have a worse outcome compared to cases showing mutated V(H) genes, that is, post-GC derived. Also, telomere length has been reported to be of prognostic significance in CLL. Interestingly, telomerase becomes activated during the GC reaction and an elongation of the telomeres occurs in GC B cells. We performed telomere length and V(H) gene analysis in a series of 61 CLL cases, in order to investigate if the unique telomere lengthening shown in GC B cells could reflect the telomere status in the two subsets of mutated and unmutated CLL. A novel association was found between V(H) gene mutation status and telomere length, since significantly shorter telomeres were demonstrated in the unmutated group compared to the mutated group (mean length 4.3 vs 6.3 kbp). Shorter telomeres also constituted a subgroup with a worse prognosis than cases with longer telomeres (median survival 59 vs 159 months). Furthermore, the Ig gene sequence data revealed that samples with high mutations frequency (>6%) had long telomeres ( approximately 8 kbp). Thus, both the telomere and V(H) gene mutation status in CLL appear linked, which may reflect the proliferative history of the clonal cells with regard to the GC reaction. PMID- 12592376 TI - DNA damage and cell cycle arrest induced by 2-(4-amino-3-methylphenyl)-5 fluorobenzothiazole (5F 203, NSC 703786) is attenuated in aryl hydrocarbon receptor deficient MCF-7 cells. AB - The fluorinated benzothiazole analogue 2-(4-amino-3-methylphenyl)-5 fluorobenzothiazole (5F 203, NSC 703786) is a novel agent with potent and selective antitumour properties and, in the form of its L-lysylamide prodrug Phortress (NSC 710305), is a current candidate for early phase clinical studies. Previous findings have indicated that cytochrome P450 1A1 (CYP1A1) may play a role in the antitumour activity of molecules in the benzothiazole series including the nonfluorinated parent compound 2-(4-amino-3 methylphenyl)benzothiazole (DF 203, NSC 674495) (Kashiyama et al, 1999; Chua et al, 2000; Loaiza-Perez et al, 2002). In this study, we assessed and verified that a fully functional aryl hydrocarbon receptor (AhR) signalling pathway is a necessary requisite for the induction of efficient cytotoxicity by 5F 203 in MCF 7 wild-type sensitive cells. Drug exposure caused MCF-7 sensitive cells to arrest in G(1) and S phase, and induced DNA adduct formation, in contrast to AhR deficient AH(R100) variant MCF-7 cells. In sensitive MCF-7 cells, induction of CYP1A1 and CYP1B1 transcription (measured by luciferase reporter assay and real time reverse transcriptase-polymerase chain reaction (RT-PCR)), and 7 ethoxyresorufin-O-deethylase (EROD) activity was demonstrated, following treatment with 5F 203. In contrast, in resistant AH(R100) cells, drug treatment did not affect CYP1A1 and CYP1B1 transcription and EROD activity. Furthermore, AH(R100) cells failed to produce either protein/DNA complexes on the xenobiotic responsive element (XRE) sequence of CYP1A1 promoter (measured by electrophoretic mobility shift assay) or DNA adducts. The data confirm that activation of the AhR signalling pathway is an important feature of the antitumour activity of 5F 203. PMID- 12592378 TI - Arginine deprivation, growth inhibition and tumour cell death: 2. Enzymatic degradation of arginine in normal and malignant cell cultures. AB - Arginase added to culture medium reduced arginine to negligible levels within approximately 6 h, and enzyme activity persisted relatively undiminished for at least 3 days. Human and bovine arginase proved equally effective. The response of normal cells was to enter G1 (G0) arrest, from which most of the cells could be recovered weeks later. In contrast, malignant cell lines treated with unpegylated or pegylated enzyme resulted in cell death on a massive scale within 3 - 5 days, with a very low to negligible percentage of cells (<0.01%) being recoverable on restoration with arginine. Although pegylation resulted in a 40% drop in specific activity, arginase was considerably more stable and remained active for >>8 days. Arginine decarboxylase caused malignant cell arrest at the same units per millilitre as arginase. Its breakdown product, agmatine, was relatively nontoxic in the presence of arginine, but exacerbated cell death above millimolar concentration in its absence. Although ornithine failed to rescue cells from deprivation, citrulline recovered cells in all cases, although less well in fast growing tumour cell populations, whereas readdition of arginine failed to work unless a complete medium change was given (because of the persistence of the enzymes in the medium catabolising its destruction). The advantages and disadvantages of these two arginine-catabolising enzymes are discussed, and compared with arginine deiminase. PMID- 12592377 TI - Gelsolin suppresses tumorigenicity through inhibiting PKC activation in a human lung cancer cell line, PC10. AB - Gelsolin expression is frequently downregulated in lung cancer and several types of different human cancers. To examine the effects of gelsolin restoration on tumorigenicity, we here stably expressed various levels of gelsolin via gene transfer in lung cancer cells (squamous cell carcinoma line, PC10). We observed the alterations in tumorigenicity in vivo when implanted in nude mice, and the changes in growth properties in vitro. As compared to parental cells and control clones, gelsolin transfectants highly reduced tumorigenicity and repressed cell proliferation. Moreover, we investigated bradykinin-induced responses in gelsolin overexpressing clones, because agonist-stimulated activation of the phospholipases C (PLC)/protein kinase C (PKC) signal transduction pathway is critical for cell growth and tumorigenicity. Bradykinin promotes phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis by PLC and translocation of various PKC isoforms from the cytosolic fraction to the particulate fraction. Bradykinin treatment did not increase inositoltriphosphate (IP3) production and induce the membrane fractions of PKC alpha and PKC gamma in gelsolin tranfectants, while it induced PIP2 hydrolysis and increased the fractions in parental and control clones. These results suggest that gelsolin suppressed the activation of PKCs involved in phospholipid signalling pathways, inhibiting cell proliferation and tumorigenicity. PMID- 12592379 TI - Constitutive nuclear factor-kappa B mRNA, protein overexpression and enhanced DNA binding activity in thymidylate synthase inhibitor-resistant tumour cells. AB - In this study, the gene copy number, mRNA and protein expression levels and nuclear DNA-binding activity of nuclear factor kappa B (NF-kappa B) were compared in a panel of five pairs of thymidylate synthase (TS) inhibitor-resistant and wild-type parent cancer cell lines. High constitutive NF-kappa B DNA-binding activity was detected in all chemoresistant cell lines. The upregulated NF-kappa B activity was composed of NF-kappa B subunits p50 and p65. Four out of five resistant cell lines constitutively overexpressed NF-kappa B p50 and p65 mRNA and protein. One resistant cell line with the highest NF-kappa B DNA-binding activity showed normal p50 and p65 protein expression. No NF-kappa B gene amplification was detected in resistant cell lines. Transient exposure of wild-type RKO(WT) and H630(WT) cells to 5-FU induced NF-kappa B DNA-binding activity but had no effect on NF-kappa B protein expression in these cells. Our results indicate that high constitutive NF-kappa B activity caused by its gene overexpression is an intrinsic character of TS inhibitor-resistant cells. NF-kappa B can antagonise anticancer drug-induced apoptosis. High NF-kappa B expression and nuclear activity in TS inhibitor-resistant cancer cells may play an important role in the chemoresistance. PMID- 12592380 TI - Inhibition of IL-6+IL-6 soluble receptor-stimulated aromatase activity by the IL 6 antagonist, Sant 7, in breast tissue-derived fibroblasts. AB - Interleukin 6 (IL-6) and its soluble receptor (IL-6sR) can markedly stimulate aromatase activity in cultured fibroblasts derived from normal or malignant breast tissues. IL-6 acts by binding to a low-affinity membrane-spanning receptor (IL-6R), which must associate with a high-affinity receptor (gp130) for signal transduction to occur. Sant 7 is a mutated form of IL-6 that can bind to the IL 6R, but inhibits its ability to interact with the gp130 signal transducing protein. In this study, we have used Sant 7 to examine its ability to inhibit IL 6+IL-6 soluble receptor (IL-6sR)-stimulated aromatase activity in breast tissue derived fibroblasts. As previously observed, IL-6+IL-6sR markedly stimulated aromatase activity (7.7-20.8-fold) in fibroblasts derived from reduction mammoplasty tissue, tissue proximal to tumours and breast tumours. Sant 7 inhibited basal aromatase activity in some fibroblasts by 25-30% that had a high basal activity, but almost completely blocked the ability of IL-6+IL-6sR to stimulate aromatase activity. The IC(50) for the inhibition of IL-6+IL-6sR stimulated aromatase activity by Sant 7 was 60 ng ml(-1). A comparison of the effects of prostaglandin E(2) (PGE(2)), which can also regulate aromatase activity, and IL-6+IL-6sR revealed a greater degree of aromatase stimulation by IL-6+IL-6sR. Sant 7, however, inhibited PGE(2)-stimulated aromatase activity by 70% suggesting that PGE(2) acts, in part, by stimulating IL-6 production. Much of the IL-6 and IL-6sR available to stimulate breast tumour aromatase activity may originate from infiltrating macrophages and lymphocytes. The ability to block aromatase stimulation by these factors may offer a novel therapeutic strategy for reducing oestrogen synthesis in breast tumours. PMID- 12592381 TI - Nuclear export inhibitor leptomycin B induces the appearance of novel forms of human Mdm2 protein. AB - The nuclear export inhibitor leptomycin B (LMB) prevents the export of proteins from the nucleus to the cytoplasm, protects p53 from Mdm2-mediated degradation and is a very potent inducer of the p53 transcriptional activity. Here we suggest that LMB can also interfere with the degradation of human Mdm2. In the presence of this drug, we observed two novel forms of this protein: a slow mobility form and an amino-terminal fragment with an apparent molecular mass of 32 kDa. The presence of this 32 kDa band is abolished with proteasome inhibitors, indicating that its appearance could be because of limited processing by the proteasome. These results may be useful in understanding the mechanism of degradation of Mdm2 by the proteasome. PMID- 12592382 TI - ERK signaling pathway is involved in p15INK4b/p16INK4a expression and HepG2 growth inhibition triggered by TPA and Saikosaponin a. AB - The signal pathway mediating induction of p15(INK4b) and p16(INK4a) during HepG2 growth inhibition triggered by the phorbol ester tumor promoter TPA (12-O tetradecanoylphorbol 13-acetate) and the Chinese herb Saikosaponin a was investigated. Western blot of three activated forms of mitogen-activated protein kinase (MAPK) (p-ERK, p-JNK and p-p38) demonstrated that phosphorylation of ERK is dramatically induced (11.6-fold ) by TPA during 15 min to 1 h and significantly induced (2.5-fold) by Saikosaponin alpha at 30 min, whereas phosphorylation of JNK was induced only by TPA during 30 min to 1 h. Phosphorylation of p38 was not induced by either drug. During this period, phosphorylation of one of the downstream transcriptional factors of MAPK cascade, ATF2, was 3.2- and 2.0-fold induced by TPA and Saikosaponin a, respectively, whereas that of another transcriptional factor, c-jun, was induced by TPA only. On the other hand, expressions of proto-oncogene c-jun, junB and c-fos were induced by TPA and Saikosaponin a during 30 min to 6 h of treatment. Pretreatment of 20 microg/ml PD98059, an inhibitor of MEK which is the upstream kinase of ERK, prevents the TPA- and Saikosaponin a-triggered HepG2 growth inhibition by 50 and 30%, respectively, accompanied by a 50 - 85% decrease of the p15(INK4b)/p16(INK4a) RNAs and proteins induced by both drugs. Inductions of c fos RNA by both drugs and c-jun phosphorylation by TPA were also significantly reduced by PD98059 pretreatment. In addition, AP-1 DNA-binding assay using nonisotopic capillary electrophoresis and laser-induced fluorescence (CE/LIF) demonstrated that the AP-1-related DNA-binding activity was significantly induced by TPA and Saikosaponin a, which can be reduced by PD98059 pretreatment. These results suggested that activation of ERK together with its downstream transcriptional machinery mediated p15(INK4b) and p16(INK4a) expression that led to HepG2 growth inhibition. PMID- 12592383 TI - Disruption of the B-cell specific transcriptional program in HHV-8 associated primary effusion lymphoma cell lines. AB - Primary effusion lymphoma (PEL) is a lymphoproliferative disease of B-cell origin that is associated with HHV-8 infection. PEL cells harbor a non-B, non-T phenotype and lack significant surface immunoglobulin (Ig) expression, a characteristic that has not been fully explained. In the present study, we demonstrate that PEL cells constitutively express interferon regulatory factor (IRF)-4, a transcription factor that regulates the activity of the immunoglobulin light-chain enhancer elements lambdaB and kappaE3' through binding to a composite Ets-IRF site. IRF-4 activity requires its physical interaction with PU.1, an Ets family member involved in the activation of genes essential for B-cell development. However, in PEL-derived B-cell lines, PU.1 expression was completely abrogated; expression of the B cell specific transcription factor Oct-2, which is known to regulate PU.1 expression, was also abolished. Moreover, the B-cell specific coactivator of octamer factors, BOB-1/OcaB, was expressed at very decreased levels in PEL cells. Ectopic expression of Oct-2 was able to fully restore PU.1 promoter activity in the PEL cell line BCBL-1, while PU.1 expression also reconstituted the activity of the lambdaB Ets-IRF site. In addition, protein levels of BSAP/Pax-5 and IRF-8/ICSBP were undetectable in PEL cells. The pattern of transcription factor ablation observed in PEL was found to be comparable to that observed in classical Hodgkin's disease-derived cell lines, which also lack B-cell-specific surface markers. These observations indicate that disruption of the B-cell-specific transcriptional program is likely to contribute to the incomplete B-cell phenotype characteristic of PEL cells. PMID- 12592385 TI - Overexpression of a protein fragment of RNA helicase A causes inhibition of endogenous BRCA1 function and defects in ploidy and cytokinesis in mammary epithelial cells. AB - The breast- and ovarian-specific tumor suppressor, BRCA1, has been implicated to function in many nuclear processes, including DNA damage repair, recombination, transcription, ubiquitination, cell cycle checkpoint enforcement, and centrosome regulation. Utilizing a previously described interaction between BRCA1 and RNA helicase A (RHA), we have developed a dominant-negative approach to block BRCA1 function in human breast epithelial cells. Overexpression of a truncated RHA peptide that can bind to the BRCA1 carboxy-terminus prevents normal BRCA1 function, such as BRCA1 association with nuclear foci following DNA damage. Overexpression of this dominant-negative protein induces pleomorphic nuclei, aberrant mitoses with extra centrosomes, and tetraploidy. This model system allows us to observe changes to mammary epithelial cells that occur acutely following loss of BRCA1 function. Furthermore, inhibition of BRCA1 via overexpressing the RHA fragment coincides with a reduction in PARP-1 protein expression, suggesting a possible mechanism for BRCA1 in the maintenance of genomic integrity. PMID- 12592384 TI - Type 1 insulin-like growth factor regulates MT1-MMP synthesis and tumor invasion via PI 3-kinase/Akt signaling. AB - The membrane type 1 matrix metalloproteinase (MT1-MMP) has been identified as a major activator of MMP-2 - a process involving the formation of a trimolecular complex with TIMP-2. We previously identified the IGF-I receptor as a positive regulator of MMP-2 synthesis. Here, we investigated the role of IGF-IR in the regulation of MT1-MMP. Highly invasive Lewis lung carcinoma subline H-59 cells express MT1-MMP and utilize it to activate their major extracellular matrix degrading proteinase-MMP-2. These cells were transiently transfected with a plasmid vector expressing a luciferase reporter gene downstream of the mouse MT1 MMP promoter. IGF-I treatment increased luciferase activity in the transfected cells by up to 10-fold and augmented endogenous MT1-MMP mRNA and protein synthesis by up to 2-3-fold, relative to controls. MT1-MMP induction and invasion were blocked by the PI 3-kinase inhibitors LY294002 and wortmannin and by rapamycin, but not by the MEK inhibitor PD98059. Overexpression of a dominant negative Akt mutant or of the tumor suppressor phosphatase and tensin homologue, PTEN, in these cells also caused a significant reduction in MT1-MMP expression and invasion. The results demonstrate that IGF-IR controls tumor cell invasion by coordinately regulating MMP-2 expression and its MT1-MMP-mediated activation and identify PI 3-kinase/Akt/mTOR signaling as critical to this regulation. PMID- 12592387 TI - CHD5, a new member of the chromodomain gene family, is preferentially expressed in the nervous system. AB - Chromatin remodeling is one of the mechanisms by which gene expression is regulated developmentally. Chromatin structure is controlled at least in part by post-translational modification of histones, as well as by chromodomain proteins. We have identified a novel gene encoding a protein with chromatin remodeling, helicase and DNA-binding motifs. This gene, called CHD5, is the fifth member of the CHD gene family identified in humans. This gene is most homologous to CHD3 and CHD4, which encode proteins that are part of the nucleosome remodeling and histone deacetylation (NuRD) complex. CHD5 is preferentially expressed in total brain, fetal brain, and cerebellum. It is also moderately expressed in the adrenal gland, but expression is undetectable in almost all other tissues examined. CHD5 maps within a small region of deletion on 1p36.3 in human neuroblastomas, a common pediatric tumor. We examined a panel of neuroblastoma cell lines for CHD5 expression, which was consistently low or undetectable in all these lines. Expression was also examined in a panel of 137 primary neuroblastomas, and low expression was highly correlated with 1p deletion, MYCN amplification, advanced stage, and unfavorable histology. These findings suggest that this gene may play a role in the development of the nervous system, and it may also play a role in the pathogenesis of neural tumors. PMID- 12592386 TI - Overexpression of Cdk6-cyclin D3 highly sensitizes cells to physical and chemical transformation. AB - Virtually all mammalian cells express two seemingly redundant cyclin-D-dependent kinases (Cdk4 and Cdk6) and three partner cyclins (D1, D2 and D3) essential for the G(1)-S transition, with predominant expression of Cdk4 and D1 in mesenchymal cells and Cdk6 and D3 in hematopoietic cells. We recently found two novel functions for Cdk6 executed in fibroblasts although unlike Cdk4 it is dispensable for their proliferation. In the rat fibroblast NRK-49F cells, oncogenic stimulation recruits Cdk6 to participate in a step of the cell cycle start that seems to be critical for anchorage-independent S-phase onset. Among the kinase-D type cyclin combinations, the Cdk6-cyclin-D3 complex has a unique ability to evade inhibition by cyclin-dependent kinase inhibitors and thereby control the cell's proliferative competence under growth-suppressive conditions. We describe here that 2-5-fold overexpression of both Cdk6 and D3 enhances by 5x10(3)-10(6) fold the susceptibility of the BALB/c3T3 and C3H10T1/2 mouse fibroblast lines to ultraviolet irradiation- as well as 3-methylcholanthrene-induced transformation. This result suggests that deregulated expression of Cdk6 and cyclinD3 may predispose cells to malignant transformation, supporting the recent finding that cyclin D3 activated by chromosomal rearrangement is the causative gene of non Hodgkin B lymphoma, in which Cdk6 is the major partner kinase. PMID- 12592388 TI - Beta-hydroxyisovalerylshikonin induces apoptosis in human leukemia cells by inhibiting the activity of a polo-like kinase 1 (PLK1). AB - beta-Hydroxyisovalerylshikonin (beta-HIVS), which was isolated from the plant, Lithospermum radix, induces apoptosis in various lines of human tumor cells. To identify genes involved in beta-HIVS-induced apoptotic process, we performed cDNA array analysis and found that beta-HIVS suppresses the expression of the gene for a polo-like kinase 1 (PLK1) that is involved in control of the cell cycle. When U937 and HL60 cells were treated with 10(-6) M beta-HIVS for 0.5 h, both the amount of PLK1 itself and the kinase activity of this enzyme were decreased. By contrast, Bcr-Abl-positive K562 cells were resistant to the induction of apoptosis by beta-HIVS and this compound did not suppress the kinase activity of PLK1 in these cells. However, simultaneous treatment of K562 cells with both beta HIVS and STI571, which selectively inhibits the protein tyrosine kinase (PTK) activity of Bcr-Abl, strongly induced apoptosis. Moreover, beta-HIVS increased the inhibitory effect of STI571 on PTK activity. Treatment of K562 cells with antisense oligodeoxynucleotides (ODNs) specific for PLK1 sensitized these cells to the beta-HIVS-induced fragmentation of DNA. These results suggest that suppression of the activity of PLK1 via inhibition of tyrosine kinase activity by beta-HIVS might play a critical role in the induction of apoptosis. PMID- 12592389 TI - Increased manganese superoxide dismutase (SOD-2) is part of the mechanism for prostate tumor suppression by Mac25/insulin-like growth factor binding-protein related protein-1. AB - Increased expression of mac25/insulin-like growth factor binding-protein related protein-1 (IGFBP-rP1) in human breast and prostate epithelial cell lines results in the suppression of tumor growth. CDNA expression array analysis revealed increased manganese superoxide dismutase (SOD-2) expression in the mac25/IGFBP rP1-transfected M12 human prostate cancer cell line compared to M12 control cells. SOD-2 has been postulated to be a tumor suppressor. SOD-2 was also increased in LNCaP cells stably transfected with mac25/IGFBP-rP1, but not in mac25/IGFBP-rP1-transfected PC-3 cells. Mac25 LNCaP cells had a marked decrease in tumor growth in nude mice compared to controls, but there was no difference in tumor growth in mac25 PC-3 cells compared to control. Phosphorylated Erk and Akt were increased in the M12 and LNCaP transfected mac25/IGFBP-rP1 cells but not PC 3 mac25. Inhibition of PI-3 kinase results in a marked decrease in viability of the M12-mac25 cells compared to M12 controls. Cells treated with H(2)O(2) result in an increase in phospho-ERK. Transfection of SOD-2 in M12 cells markedly decreased tumor growth, apoptosis, G1 delay in the cell cycle, and expression of senescence associated beta-galactosidase. These results suggest that one of the downstream mediators of the senescence-associated tumor suppression effect of mac25/IGFBP-rP1 is SOD-2. PMID- 12592390 TI - Inhibition of ultraviolet B-mediated activation of nuclear factor kappaB in normal human epidermal keratinocytes by green tea Constituent (-) epigallocatechin-3-gallate. AB - Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, possesses significant anti-inflammatory and cancer chemopreventive properties. Studies have shown the photochemopreventive effects of green tea and EGCG in cell culture, animal models, and human skin. The molecular mechanism(s) of photochemopreventive effects of EGCG are incompletely understood. We recently showed that EGCG treatment of the normal human epidermal keratinocytes (NHEK) inhibits ultraviolet (UV)B-mediated activation of the mitogen-activated protein kinase (MAPK) pathway. In this study, we evaluated the effect of EGCG on UVB-mediated modulation of the nuclear factor kappa B (NF-kappaB) pathway, which is known to play a critical role in a variety of physiological functions and is involved in inflammation and development of cancer. Immunoblot analysis demonstrated that the treatment of NHEK with EGCG (10-40 microM) for 24 h resulted in a significant inhibition of UVB (40 mJ/cm(2))-mediated degradation and phosphorylation of IkappaBalpha and activation of IKKalpha, in a dose-dependent manner. UVB-mediated degradation and phosphorylation of IkappaBalpha and activation of IKKalpha was also observed in a time-dependent protocol (15 and 30 min, 1, 2, 3, 6, 12 h post-UVB exposure). Employing immunoblot analysis, enzyme-linked immunosorbent assay, and gel shift assay, we demonstrate that EGCG treatment of the cells resulted in a significant dose- and time-dependent inhibition of UVB-mediated activation and nuclear translocation of a NF-kappaB/p65. Our data suggest that EGCG protects against the adverse effects of UV radiation via modulations in NF-kappaB pathway, and provide a molecular basis for the photochemopreventive effect of EGCG. PMID- 12592391 TI - IRF-1 reverts the transformed phenotype of oncogenically transformed cells in vitro and in vivo. AB - The expression of the transcriptional activator and tumor suppressor IRF-1 induces multiple effects that counteract the growth of tumor cells in vitro and in vivo. These include the inhibition of cell proliferation, the secretion of interferon-beta (IFN-beta), the induction of apoptosis specifically in certain cell types and the induction of a strong T-cell response. Here, we show that apart from its immune-activating properties, IRF-1 expression leads to a reversion of the tumorigenic phenotype of NIH3T3 cells transformed by different oncogenes. This was analysed in detail in a cell line in which the expression of c-Ha-ras and c-myc is under the control of a doxycycline-regulated promoter allowing to switch between the normal and oncogenic cell status. In the same cells, a beta-estradiol activatable IRF-1 fusion protein is expressed. After IRF 1 activation the oncogene-mediated acceleration of the cell cycle is reverted. Further, a complete IRF-1-mediated reversion of the oncogenic phenotype is observed in soft-agar growth assays. IRF-1 activation induces IFN-beta secretion; however, the observed effects are not mediated by IFN-beta. Inhibition of tumor growth is observed in nude mice as long as IRF-1 is active, indicating that neither B- nor T-cells must become activated for tumor growth suppression. PMID- 12592393 TI - Methylation of CpG dinucleotides and/or CCWGG motifs at the promoter of TP53 correlates with decreased gene expression in a subset of acute lymphoblastic leukemia patients. AB - It has been shown that methylation of CpG dinucleotides located in the promoter region of TP53 is associated with low expression levels of this gene. We have analysed the methylation status of one CpG dinucleotide and of three CCWGG motifs, also located in the promoter region of the gene, in bone marrow samples obtained from patients with acute lymphoblastic leukemia (ALL). Eight out of 25 samples analysed showed methylation of either the CpG dinucleotide, the CCWGG motifs or both. Relative to nonmethylated leukemia samples, TP53 expression levels were decreased in all methylated samples in which TP53 expression could be measured. Methylation of CpG and CCWGG motifs in the promoter of TP53 could represent a novel mechanism leading to functional impairment of this tumor suppressor gene in ALL. PMID- 12592392 TI - An extended bipartite nuclear localization signal in Smad4 is required for its nuclear import and transcriptional activity. AB - Smad proteins are a class of tumor suppressors that play critical roles in inhibiting the proliferation of a variety of cell types by modulating the transcriptions of target genes. Despite recent advances, the mechanism of their nuclear import is not completely understood. Smad proteins contain a conserved basic motif in their N-terminal MH1 domains that resembles a nuclear localization signal (NLS). Previous studies indicate that in receptor-regulated Smads such as Smad1 and Smad3 this motif determines their interactions with nuclear import receptors and mediates their ligand-induced nuclear translocation. Common-Smads such as Smad4 display constant nucleocytoplasmic shuttling and are capable of autonomous nuclear import and export. Mutations of the basic motif in Smad4 disrupted its nuclear accumulation. However, this motif is not sufficient to confer nuclear translocation to a fused heterologous protein, suggesting that it is only part of the bona fide Smad4 NLS. We mapped the Smad4 NLS by fusing various segments of Smad4 sequence covering the basic motif to GFP and tested the localization of the fusion proteins. We identified an extended NLS, starting from the basic motif and extending into the DNA-binding region (AA 45-110), that is sufficient to confer nuclear localization to GFP. Among the 14 basic residues in the NLS, only four (K45, K46, K48 and R81) are critical for import. This NLS is critical not only for autonomous nuclear import of Smad4, but also for its nuclear translocation in the presence of activated R-Smads, further confirming the functional relevance of the Smad4 NLS in TGF-beta signal transduction. Structural modeling demonstrated that the four critical basic residues are all solvent exposed and map to a single localized segment on one surface of the Smad4 MH1 domain. Their distribution and spacing resemble a classical bipartite NLS. Smad4 displays specific binding to importin alpha through its MH1 domain, which was abrogated by loss-of-function mutations in Smad4 NLS. Finally, the Smad4 NLS is essential for its transcriptional activity since loss-of-function NLS mutants are also transcriptionally inactive. PMID- 12592394 TI - Functional genomic analysis reveals distinct neoplastic phenotypes associated with c-myb mutation in the bursa of Fabricius. AB - Avian retroviral integration into the c-myb locus is casually associated with the development of lymphomas in the bursa of Farbricius of chickens; these arise with a shorter latency than bursal lymphomas caused by deregulation of c-myc. This study indicates that c-myb mutation in embryonic bursal precursors leads to an oligoclonal population of developing bursal follicles, showing a variable propensity to form a novel lesion, the neoplastic follicle (NF). About half of such bursas rapidly developed lymphomas. Detection of changes in gene expression, during the development of neoplasms, was carried out by cDNA microarray analysis. The transcriptional signature of lymphomas with mutant c-myb was more limited than, and only partially shared with, those of bursal lymphomas caused by Myc or Rel oncogenes. The c-myb-associated lymphomas frequently showed overexpression of c-myc and altered expression of other genes involved in cell cycle control and proliferation-related signal transduction. Oligoclonal, NF-containing bursas lacked detectable c-myc overexpression and demonstrated a pattern of gene expression distinct from that of normal bursa and partially shared with the short latency lymphomas. This functional genomic analysis uncovered several different pathways of lymphomagenesis by oncogenic transcription factors acting in a B-cell lineage. PMID- 12592395 TI - Cloning and molecular characterization of a novel gene strongly induced by the adenovirus E1A gene in rat thyroid cells. AB - Expression of the adenovirus E1A gene in the rat thyroid differentiated cell line PC Cl 3 induces thyrotropin-independent cell growth and impairs differentiation. However, the malignant phenotype is achieved only when the PC E1A cells are infected with other murine retroviruses carrying the v-abl, v-raf or polyoma middle-T genes. To determine through which genes E1A affects thyroid cells, we differentially screened PC Cl 3 and PC E1A cells. Here we report a new gene, named CL2, that is upregulated in PC E1A cells. The CL2 transcript is 4.4 kb long and encodes a 949 amino-acid protein. Conceptual translation of the open reading frame showed one product with a signal peptide, multiple nuclear localization signals and three newly described domains. Furthermore, in vivo, this protein was located juxtanuclear, which is suggestive of Golgian localization, and also in cytoplasm and nucleus/nucleolus. Finally, CL2 gene expression was drastically downregulated in human thyroid neoplastic cell lines and tissues. PMID- 12592396 TI - Mapping of genetic modifiers of thymic lymphoma development in p53-knockout mice. AB - The strain dependency of the spectrum and latency of tumors has been reported in p53-deficient (KO) mice, suggesting the presence of modifiers for the outcome of the p53 deficiency. The modifiers provide clues to the oncogenic pathway in cells lacking p53, the most frequently mutated gene in a wide variety of human cancers. To search the modifiers, we induced 160 lymphomas and 69 skin tumors by gamma irradiation of p53(KO/+) backcross mice between BALB/c and MSM strains and performed genome scan. BALB/c-derived alleles at three loci on chromosome 19, Mp53D1 (modifier of p53-deficiency) at D19Mit5, Mp53D2 at D19Mit90 and Mp53D3 at D19Mit123, extended the latency of thymic lymphoma development (P values in Mantel-Cox test were 0.0007, 0.0007 and 0.0003, respectively). Mp53D3 also increased the latency of skin tumors (P value, 0.0008). The linkage of Mp53D2 was confirmed by the experiment using 94 p53-KO mice consomic for chromosome 19, providing a significant linkage. However, the linkage was not confirmed for Mp53D1 or Mp53D3, suggesting epistasis of genes involved in the tumorigenesis. PMID- 12592397 TI - Genomic characterization of the chromosomal breakpoints of t(4;14) of multiple myeloma suggests more than one possible aetiological mechanism. AB - Using FISH-based techniques, rearrangements of the immunoglobulin heavy-chain (IgH) locus at 14q32 have been found in the majority of cases of multiple myeloma (MM). Some of these IgH translocations are recurrent and we have characterized the genomic breakpoints of seven t(4;14) translocations from MM patients, using a combination of vectorette and conventional polymerase chain reaction methods, the aim being to understand the molecular mechanism leading to MM. Conventionally, the chromosome 14q32 breakpoints in these reciprocal translocations are believed to be located in the IgH mu switch (S) region and a further downstream S region with deletion of intervening DNA occurring as a result of aberrant class switch recombination (CSR); this was seen in five of the cases analysed. However, in two patients it was possible to demonstrate that the rearranged hybrid switch region sequence was joined to DNA from chromosome 4p16, suggesting that IgH translocations can occur in B cells that have already undergone legitimate CSR. The complex nature of these rearrangements leads us to speculate that primary IgH translocations may occur at different time points in the development in MM plasma cells, either at the time of physiological CSR or at a later stage, possibly involving a different mechanism. PMID- 12592398 TI - Product-assisted catalysis in base-excision DNA repair. AB - Most spontaneous damage to bases in DNA is corrected through the action of the base-excision DNA repair pathway. Base excision repair is initiated by DNA glycosylases, lesion-specific enzymes that intercept aberrant bases in DNA and catalyze their excision. How such proteins accomplish the feat of catalyzing no fewer than five sequential reaction steps using a single active site has been unknown. To help answer this, we report the structure of a trapped catalytic intermediate in DNA repair by human 8-oxoguanine DNA glycosylase. This structure and supporting biochemical results reveal that the enzyme sequesters the excised lesion base and exploits it as a cofactor to participate in catalysis. To our knowledge, the present example represents the first documented case of product assisted catalysis in an enzyme-catalyzed reaction. PMID- 12592399 TI - The H-NS dimerization domain defines a new fold contributing to DNA recognition. AB - H-NS, a protein found in Gram-negative bacteria, is involved in structuring the bacterial chromosome and acts as a global regulator for the expression of a wide variety of genes. These functions are correlated with both its DNA-binding and oligomerization properties. We have identified the minimal dimerization domain of H-NS, a 46 amino acid-long N-terminal fragment, and determined its structure using heteronuclear NMR spectroscopy. The highly intertwined structure of the dimer, reminiscent of a handshake, defines a new structural fold, which may offer a possibility for discriminating prokaryotic from eukaryotic proteins in drug design. Using mutational analysis, we also show that this N-terminal domain actively contributes to DNA binding, conversely to the current paradigm. Together, our data allows us to propose a model for the action of full length H NS. PMID- 12592400 TI - A novel viral mechanism for dysregulation of beta-catenin in Kaposi's sarcoma associated herpesvirus latency. AB - The Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is expressed in all KSHV-associated tumors, including Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL). We found that beta-catenin is overexpressed in both PEL cells and KS tissue. Introduction of anti-LANA small interfering RNA (siRNA) into PEL cells eliminated beta-catenin accumulation; LANA itself upregulated expression of beta-catenin in transfected cells. LANA stabilizes beta-catenin by binding to the negative regulator GSK-3beta, causing a cell cycle-dependent nuclear accumulation of GSK-3beta. The LANA C terminus contains sequences similar to the GSK-3beta-binding domain of Axin. Disruption of this region resulted in a mutant LANA that failed to re-localize GSK-3beta or stabilize beta-catenin. The importance of this pathway to KSHV-driven cell proliferation was highlighted by the observation that LANA, but not mutant LANA, stimulates entry into S phase. Redistribution of GSK-3beta can therefore be a source of beta-catenin dysregulation in human cancers. PMID- 12592401 TI - Epstein-Barr virus infection of polarized tongue and nasopharyngeal epithelial cells. AB - Epstein-Barr virus (EBV) initially enters the body through the oropharyngeal mucosa and subsequently infects B lymphocytes through their CD21 (CR2) complement receptor. Mechanisms of EBV entry into and release from epithelial cells are poorly understood. To study EBV infection in mucosal oropharyngeal epithelial cells, we established human polarized tongue and pharyngeal epithelial cells in culture. We show that EBV enters these cells through three CD21-independent pathways: (i) by direct cell-to-cell contact of apical cell membranes with EBV infected lymphocytes; (ii) by entry of cell-free virions through basolateral membranes, mediated in part through an interaction between beta1 or alpha5beta1 integrins and the EBV BMRF-2 protein; and (iii) after initial infection, by virus spread directly across lateral membranes to adjacent epithelial cells. Release of progeny virions from polarized cells occurs from both their apical and basolateral membranes. These data indicate that multiple approaches to prevention of epithelial infection with EBV will be necessary. PMID- 12592402 TI - Toll-like receptor-4 (TLR4) signaling augments chemokine-induced neutrophil migration by modulating cell surface expression of chemokine receptors. AB - Polymorphonuclear leukocytes (PMNs) are critical effector cells of the innate immune system that protect the host by migrating to inflammatory sites and killing pathogenic microbes. We addressed the role of chemokine receptor desensitization induced by G-protein-coupled receptor kinases (GRKs) in the feedback control of PMN migration. We show that the chemokine macrophage inflammatory protein-2 (MIP-2) induces GRK2 and GRK5 expression in PMNs through phosphoinositide-3-kinase (PI3K)-gamma signaling. We also show that lipopolysaccharide (LPS)-activated signaling through the Toll-like receptor (TLR) 4 pathway transcriptionally downregulates the expression of GRK2 and GRK5 in response to MIP-2. The reduced expression of GRKs lowers chemokine receptor desensitization and markedly augments the PMN migratory response. These data indicate that TLR4 modulation of PMN surface chemokine receptor expression subsequent to the downregulation of GRK2 and GRK5 expression is a critical determinant of PMN migration. PMID- 12592403 TI - Benfotiamine blocks three major pathways of hyperglycemic damage and prevents experimental diabetic retinopathy. AB - Three of the major biochemical pathways implicated in the pathogenesis of hyperglycemia induced vascular damage (the hexosamine pathway, the advanced glycation end product (AGE) formation pathway and the diacylglycerol (DAG) protein kinase C (PKC) pathway) are activated by increased availability of the glycolytic metabolites glyceraldehyde-3-phosphate and fructose-6-phosphate. We have discovered that the lipid-soluble thiamine derivative benfotiamine can inhibit these three pathways, as well as hyperglycemia-associated NF-kappaB activation, by activating the pentose phosphate pathway enzyme transketolase, which converts glyceraldehyde-3-phosphate and fructose-6-phosphate into pentose-5 phosphates and other sugars. In retinas of diabetic animals, benfotiamine treatment inhibited these three pathways and NF-kappaB activation by activating transketolase, and also prevented experimental diabetic retinopathy. The ability of benfotiamine to inhibit three major pathways simultaneously might be clinically useful in preventing the development and progression of diabetic complications. PMID- 12592404 TI - Neural mechanisms of general fluid intelligence. AB - We used an individual-differences approach to test whether general fluid intelligence (gF) is mediated by brain regions that support attentional (executive) control, including subregions of the prefrontal cortex. Forty-eight participants first completed a standard measure of gF (Raven's Advanced Progressive Matrices). They then performed verbal and nonverbal versions of a challenging working-memory task (three-back) while their brain activity was measured using functional magnetic resonance imaging (fMRI). Trials within the three-back task varied greatly in the demand for attentional control because of differences in trial-to-trial interference. On high-interference trials specifically, participants with higher gF were more accurate and had greater event-related neural activity in several brain regions. Multiple regression analyses indicated that lateral prefrontal and parietal regions may mediate the relation between ability (gF) and performance (accuracy despite interference), providing constraints on the neural mechanisms that support gF. PMID- 12592405 TI - Dynamic balance of metabotropic inputs causes dorsal horn neurons to switch functional states. AB - Sensory relay structures in the spinal cord dorsal horn are now thought to be active processing structures that function before supraspinal sensory integration. Dorsal horn neurons directly receive nociceptive (pain) signals from the periphery, express a high degree of functional plasticity and are involved in long-term sensitization and chronic pain. We show here that deep dorsal horn neurons (DHNs) in Wistar rats can switch their intrinsic firing properties from tonic to plateau or endogenous bursting patterns, depending upon the balance of control by metabotropic glutamate (mGlu) and GABA(B) receptors. We further show that this modulation acts on at least one common target, the inwardly rectifying potassium channel (Kir3). Finally, we found that these firing modes correspond to specific functional states of information transfer in which dorsal horn neurons can faithfully transmit, greatly enhance or block the transfer of nociceptive information. PMID- 12592406 TI - Robo and Frazzled/DCC mediate dendritic guidance at the CNS midline. AB - Neuronal connectivity is established by the axo-dendritic polarity, correct guidance and targeting of neurons. Unlike for axons, the mechanisms responsible for directed outgrowth of dendrites are not well understood. Using single-cell labeling, we describe specific guidance defects in dendrites of identified neurons in frazzled, robo, netrin and commissureless mutant embryos of Drosophila melanogaster. We found that the cell-surface molecules Frazzled and Robo work as guidance molecules not only for axons but also for dendrites as they navigate within the CNS. Furthermore, we report that each neuron showed a cell-autonomous and independent use of guidance molecules. PMID- 12592407 TI - An activity-dependent switch to cap-independent translation triggered by eIF4E dephosphorylation. PMID- 12592408 TI - Kv3.4 subunits enhance the repolarizing efficiency of Kv3.1 channels in fast spiking neurons. AB - Neurons with the capacity to discharge at high rates--'fast-spiking' (FS) neurons -are critical participants in central motor and sensory circuits. It is widely accepted that K+ channels with Kv3.1 or Kv3.2 subunits underlie fast, delayed rectifier (DR) currents that endow neurons with this FS ability. Expression of these subunits in heterologous systems, however, yields channels that open at more depolarized potentials than do native Kv3 family channels, suggesting that they differ. One possibility is that native channels incorporate a subunit that modifies gating. Molecular, electrophysiological and pharmacological studies reported here suggest that a splice variant of the Kv3.4 subunit coassembles with Kv3.1 subunits in rat brain FS neurons. Coassembly enhances the spike repolarizing efficiency of the channels, thereby reducing spike duration and enabling higher repetitive spike rates. These results suggest that manipulation of K3.4 subunit expression could be a useful means of controlling the dynamic range of FS neurons. PMID- 12592409 TI - An evolutionarily conserved dileucine motif in Shal K+ channels mediates dendritic targeting. AB - The molecular mechanisms underlying polarized sorting of proteins in neurons are poorly understood. Here we report the identification of a 16 amino-acid, dileucine-containing motif that mediates dendritic targeting in a variety of neuronal cell types in slices of rat brain. This motif is present in the carboxy (C) termini of Shal-family K+ channels and is highly conserved from C. elegans to humans. It is necessary for dendritic targeting of potassium channel Kv4.2 and is sufficient to target the axonally localized channels Kv1.3 and Kv1.4 to the dendrites. It can also mediate dendritic targeting of a non-channel protein, CD8. PMID- 12592410 TI - Recombinant Dicer efficiently converts large dsRNAs into siRNAs suitable for gene silencing. AB - RNA interference (RNAi) is a powerful method for specifically silencing gene expression in diverse cell types. RNAi is mediated by approximately 21-nucleotide small interfering RNAs (siRNAs), which are produced from larger double-stranded RNAs (dsRNAs) in vivo through the action of Dicer, an RNase III-family enzyme. Transfecting cells with siRNAs rather than larger dsRNAs avoids the nonspecific gene silencing of the interferon response, underscoring the importance of developing efficient methods for producing reliable siRNAs. Here we show that pools of 20- to 21-base pair (bp) siRNAs can be produced enzymatically in vitro using active recombinant Dicer. Yields of < or = 70% are obtained, and the siRNAs can be easily separated from any residual large dsRNA by a series of spin columns or gel purification. Dicer-generated siRNAs (d-siRNAs) are effective in silencing transiently transfected reporter genes and endogenous genes, making in vitro dicing a useful, practical alternative for the production of siRNAs. PMID- 12592411 TI - Characterization of the human heart mitochondrial proteome. AB - To gain a better understanding of the critical role of mitochondria in cell function, we have compiled an extensive catalogue of the mitochondrial proteome using highly purified mitochondria from normal human heart tissue. Sucrose gradient centrifugation was employed to partially resolve protein complexes whose individual protein components were separated by one-dimensional PAGE. Total in gel processing and subsequent detection by mass spectrometry and rigorous bioinformatic analysis yielded a total of 615 distinct protein identifications. All protein pI values, molecular weight ranges, and hydrophobicities were represented. The coverage of the known subunits of the oxidative phosphorylation machinery within the inner mitochondrial membrane was >90%. A significant proportion of identified proteins are involved in signaling, RNA, DNA, and protein synthesis, ion transport, and lipid metabolism. The biochemical roles of 19% of the identified proteins have not been defined. This database of proteins provides a comprehensive resource for the discovery of novel mitochondrial functions and pathways. PMID- 12592412 TI - Scanning the human genome with combinatorial transcription factor libraries. AB - Despite the critical importance of transcription factors in mediating gene regulation, there exists no general, genome-wide tool that uses transcription factors to induce or silence a target gene or select for a particular phenotype. In the strategy described here, we prepared large combinatorial libraries of artificial transcription factors comprising three or six zinc-finger domains, and selected transcription factor-DNA interactions able to upregulate several genes in human cells. Selected transcription factors either induced the expression of an endothelial-specific differentiation marker, VE-cadherin, in non-endothelial cell lines or, when combined with a repression domain, knocked down expression. Potential binding sites for a number of these transcription factors were mapped along the promoter of CDH5, the gene encoding VE-cadherin. Transcription factor libraries represent a useful approach for studying and modulating gene function in cells and potentially in whole organisms. PMID- 12592413 TI - Human zinc fingers as building blocks in the construction of artificial transcription factors. AB - We describe methods for generating artificial transcription factors capable of up or downregulating the expression of genes whose promoter regions contain the target DNA sequences. To accomplish this, we screened zinc fingers derived from sequences in the human genome and isolated 56 zinc fingers with diverse DNA binding specificities. We used these zinc fingers as modular building blocks in the construction of novel, sequence-specific DNA-binding proteins. Fusion of these zinc-finger proteins with either a transcriptional activation or repression domain yielded potent transcriptional activators or repressors, respectively. These results show that the human genome encodes zinc fingers with diverse DNA binding specificities and that these domains can be used to design sequence specific DNA-binding proteins and artificial transcription factors. PMID- 12592414 TI - NTP technical report on the toxicity studies of p-tert-butylcatechol (CAS No. 98 29-3) administered in feed to F344/N rats and B6C3F1 mice. AB - [molecular structure: see text] p-tert-Butylcatechol is used as an antioxidant, stabilizer, and polymerization inhibitor for styrene, butadiene, neoprene, and other olefins and reactive monomers. p-tert-Butylcatechol was nominated by the National Cancer Institute and the U.S. Food and Drug Administration for testing based on reports of its increasing levels of production and use and to compare the toxicity of p-tert-butylcatechol with that of similar antioxidants, butylated hydroxyanisole and butylated hydroxytoluene, which are added to food. Male and female F344/N rats and B6C3F1 mice were exposed to p-tert-butylcatechol (greater than 99% pure) in feed for 15 days or 14 weeks. Genetic toxicology studies were conducted in Salmonella typhimurium, rat bone marrow cells, and mouse peripheral blood erythrocytes. In the 15-day studies, groups of five male and five female rats and mice were fed diets containing 0, 3,125, 6,250, 12,500, 25,000, or 50,000 ppm p-tert-butylcatechol (equivalent to average daily doses of approximately 290 to 2,470 mg p-tert-butylcatechol/kg body weight to rats and 590 to 8,200 mg/kg to mice). All animals in the 50,000 ppm groups were killed moribund on day 8 (rats) or by day 7 (mice). Mean body weights of all groups of rats exposed to 6,250 ppm or greater were significantly less than those of the controls. Mean body weights of male mice exposed to 12,500 or 25,000 ppm and of 25,000 ppm female mice were significantly less than those of the controls. Female rats, male and female mice in the 25,000 ppm groups, and 12,500 ppm male mice lost weight during the studies. Feed consumption by exposed rats generally decreased with increasing exposure concentration; feed consumption by exposed mice was similar to that by the controls. Thymus weights of 25,000 ppm rats and mice were significantly less than those of the controls. Gross findings noted at necropsy included thin carcasses for three male and all female rats in the 12,500 ppm groups and all male and female rats and mice in the 25,000 and 50,000 ppm groups. No exposure-related lesions were observed microscopically. In the 14-week studies, groups of 10 male and 10 female rats and mice were fed diets containing 0, 781, 1,562, 3,125, 6,250, or 12,500 ppm p-tert-butylcatechol (equivalent to average daily doses of approximately 70 to 1,030 mg/kg to rats and 135 to 2,815 mg/kg to mice). All animals survived to the end of the studies. Mean body weights of male rats exposed to 1,562 ppm or greater, female rats exposed to 3,125 ppm or greater, male mice exposed to 12,500 ppm, and female mice exposed to 6,250 or 12,500 ppm were significantly less than those of the controls. Feed consumption by male and female rats in the 6,250 and 12,500 ppm groups at week 1 and the 12,500 ppm groups at week 14 was less than that by the controls; feed consumption by exposed and control mice was similar. An erythrocytosis, indicated by increased hematocrit values, hemoglobin concentrations, and erythrocyte counts, was observed in 6,250 and 12,500 ppm rats on day 4 and in 12,500 ppm rats on day 22. At these time points, a transient hepatic effect was demonstrated by increases in alanine aminotransferase activities and bile salt concentrations in exposed rats. In 12,500 ppm male rats, absolute left cauda epididymis, epididymis, and testis weights were decreased by 15%, 10%, and 9%, respectively, compared to the controls. The number of spermatid heads per testis and epididymal sperm motility of male rats in the 12,500 ppm group were significantly less than those of the controls. The numbers of cycling female rats and females with regular estrous cycles were decreased in the 6,250 and 12,500 ppm groups. Exposed groups of females had significantly fewer estrous cycles than did the controls. Estrous cycle length increased with increasing exposure concentration; female rats in the 6,250 and 12,500 ppm groups had significantly longer cycles and spent more time in diestrus and less time in proestrus, estrus, and metestrus than did the controls. Female mice in the 12,500 ppm group had a significantly longer estrous cycle than did the controls. The incidences of hyperkeratosis of the forestomach epithelium were significantly increased in male and female rats in all exposed groups and in 12,500 ppm female mice. The incidences of hyperplasia of the forestomach epithelium were significantly increased in male and female rats exposed to 3,125 ppm or greater, male mice exposed to 12,500 ppm, and female mice exposed to 6,250 or 12,500 ppm. The severities of the forestomach lesions were minimal to moderate in male rats and minimal to mild in female rats and in mice. All male rats exposed to 6,250 or 12,500 ppm had minimal cytoplasmic alteration in the liver. The absorption, distribution, metabolism, and excretion of p-tert-butylcatechol following intravenous injection, gavage dosing, or dermal application were determined in male F344/N rats and B6C3F1 mice. The absorption of [(14)C]-p-tert-butylcatechol following gavage dosing or dermal application was high. The percent absorption following dermal application increased with increasing dose. Peak concentrations of [(14)C]-p-tert-butylcatechol equivalents in plasma were reached 1 hour after gavage dosing (200 mg/kg) and 2 hours after dermal application (60 mg/kg); no parent compound was detected in the plasma extracts. Regardless of route of administration, p-tert-butylcatechol derived radioactivity was readily excreted in the urine and was markedly nonpersistent in the tissues. p-tert- Butylcatechol was excreted as p-tert-butylcatechol sulfate and other polar metabolites that included predominately sulfate conjugates; it was not excreted as the parent compound. One metabolite was determined to be an O methyl- ON-sulfate of p-tert-butylcatechol. p-tert-Butylcatechol (10 to 1,000 microg/plate) was not mutagenic in any of several strains of S. typhimurium with or without rat or hamster liver S9. Bone marrow micronucleus tests in which 125 to 500 mg/kg p-tert-butylcatechol was administered three times by intraperitoneal injection to male rats gave negative results. No increases in the frequencies of micronucleated normochromatic erythrocytes were observed in the peripheral blood of male or female mice administered p-tert-butylcatechol in feed for 14 weeks. No significant alteration in the percentage of polychromatic erythrocytes in mouse bone marrow was observed. In summary, the primary toxicity of p-tert butylcatechol was to the forestomach of rats and mice. In the 14-week study in rats, forestomach toxicity was observed at all exposure concentrations, and the no-observed-adverse-effect level (NOAEL) was not reached for this effect. In the 14-week study in mice, the NOAEL for forestomach toxicity was 1,562 ppm. PMID- 12592415 TI - Diagnosis and management of unexplained infertility: an update. AB - Unexplained infertility constitutes around 15% of patients presenting with infertility. A lack of agreement exists among infertility specialists with regard to the diagnostic tests to be performed and their prognostic value as well as criteria of normality. It seems that serum progesterone for detection of ovulation, hysterosalpingography and or laparoscopy for tubal patency and semen analysis are the basic tests for diagnosis of unexplained infertility. Expectant treatment is the option of choice for young patients with short period of infertility. The spontaneous pregnancy rate is very high in this group of patients. The world literature have shown that controlled ovarian hyperstimulation and intrauterine insemination (COH and IUI) is an effective treatment of unexplained infertility. According to the available data, this procedure could be limited to three trials. There is evidence that both COH and IUI are important independent positive factors in achieving better pregnancy rate in unexplained infertility. If the above measures fail to achieve pregnancy, GIFT or IVF/ICSI could be performed as it yields a high pregnancy rate. PMID- 12592416 TI - Circulating biomarkers of oxidative stress in complicated pregnancies. AB - Increased lipid peroxidation (LPO) and reduced antioxidant activity may contribute to the development of complications in pregnancy. The present study discusses the possibility of LPO and antioxidant activity in both maternal and umbilical cord blood as an indicator of oxygen radical activity. For this aim, pregnancies with hypertension and pre-eclampsia, diabetes mellitus (insulin dependent diabetes mellitus and gestational diabetes mellitus), oligohydramnios and abruptio placentae, as well as a healthy control group, were subjected in the present study. Simultaneous determination of glutathione S-transferase (GST), selenium dependent glutathione peroxidase (Se-GPx), catalase (CAT) activities and thiobarbituric acid reactive-substances (TBARs) levels were carried out in maternal erythrocyte and plasma in the antenatal period (in the third trimester) and immediately after the delivery. The same oxidative stress-related parameters were determined in umbilical cord blood as well. Erythrocyte GST activity was significantly increased in insulin-dependent diabetic pregnancy (IDDP) when compared to the control (P<0.05). Erythrocyte Se-GPx activity was found to be significantly increased in hypertensive preeclamptic pregnancy (HPP) (P<0.05) and in IDDP (P<0.05). Alterations in enzyme activities were accompanied by a simultaneous significant increase in the levels of TBARs in plasma samples of HPP (P<0.05), and IDDP (P<0.05). Enzyme activities were found to be significantly lower in cord blood samples than the maternal values, except GST. This enzyme represents about two- to threefold higher activity than those of the maternal activity in uncomplicated and complicated groups. Cord blood erythrocyte and plasma Se-GPx and CAT activities were decreased significantly in the HPP group when compared to the maternal value (P<0.05). Cord blood erythrocyte CAT activity was significantly decreased in the HPP group compared to the control (P<0.05). Cord blood TBARs levels were significantly lower than the before deliveries maternal value in the HPP group (P<0.05). No difference was detected between umbilical cord blood and maternal blood TBARs levels after delivery. The results of the present study suggest that oxidative stress and subsequent lipid peroxidation accompany the complications of hypertension, preeclampsia and diabetes mellitus in pregnancy. Maternal erythrocyte GST activity seems to be a sensitive indicator of oxidative stress in IDDP before delivery. The same enzyme can be used in cord blood as a biomarker of oxidative stress upon a sudden increase in oxygenation during delivery. These multiparameter biomarkers can also be used in monitoring the efficiency of antioxidant supplementation in complicated pregnant women, as has recently been suggested for diabetic and preeclamptic pregnancies. PMID- 12592417 TI - Surveillance study of Sinupret in comparison with data of the Mainz birth registry. AB - In about 80% of all pregnant women medical drugs are used during pregnancy. Phytomedicines play an important role in self- and prescribed medication. Since the thalidomide tragedy teratogenic effects of medicamentous treatment are well known. Only a few investigations and studies on the tolerability and safety of plant-based medicines in pregnancy have been published. From 1992-1997 a nationwide retrospective surveillance study of 762 pregnant women (786 newborns) was conducted to evaluate the safety of the herbal combination preparation Sinupret during pregnancy. Standardized questionnaires were used for retrospective and systematic examination of all pregnancies. All cases were treated with Sinupret for at least 24 hours. The data of the population-based Mainz birth registry were used for comparison, interpretation and evaluation of the Sinupret study. For infants with birth defects and/or clinically abnormal pregnancies single-case-analyses were performed. The study collective was representative for Germany. 13 single-case-analyses were performed. In three cases the pregnancy ended with an abortion and in one case with an intrauterine death of the fetus. Nine newborns (1.1%) were diagnosed with major malformations. The critical evaluation of the collective and the individual cases yielded no evidence of possible teratogenic and/or embryotoxic effects of the phytomedicine in 11 out of 13 cases. In the remaining two cases a causal relationship is theoretically possible but very unlikely. On the whole the study underlines the value of the pediatric epidemiologic research in malformations of the Mainz birth register for the monitoring, assessment and evaluation of pharmaceutical studies on drug safety during pregnancy. PMID- 12592418 TI - Study of the third stage of labor by color Doppler sonography. AB - We wanted to evaluate whether improvement in ultrasound equipment in the last 5 years altered our perception of the phases of placental separation during the third stage of labor. We also investigated the influence of active management on the third stage of labor after sonographically verified placental separation. Between January and November 2001, the third stage of labor was examined in 55 women at 37-41 weeks of gestation by color Doppler sonography. The duration of blood flow between the myometrium and the placenta, the latent phase, the detachment phase, and the expulsion phase were measured and compared with the corresponding values of an earlier cohort of 57 patients investigated between November 1994 and August 1995. In the later cohort, both the duration of maternal blood flow and the detachment phase were significantly longer than in the earlier cohort (33 s +/- 48 s vs 0 s, P<0.0001 and 56 s+/-45 s vs 37 s+/-21 s, P<0.01, respectively), whereas the latent phase was significantly shorter (101 s+/-87 s vs 213 s+/-180 s, P<0.0001). There was no statistically significant difference in the length of the expulsion phase or the third stage of labor. The later cohort showed a statistically significantly more frequent multiphasic placental detachment ( P<0.05). Improvement in ultrasound equipment resulted in an earlier detection of the onset of placental separation, leading to a shorter latent phase and consecutively increased duration of the detachment phase, whereas the total duration of the third stage of labor remained unchanged. Furthermore, increased sensitivity of Doppler sonography led to a longer visualization of blood flow between the myometrium and the placenta in the normal third stage of labor. PMID- 12592419 TI - Serum adenosine deaminase activity and its isoenzyme pattern in women with normal pregnancies. AB - Adenosine deaminase (ADA) is a purine enzyme which is essential for the proliferation, maturation and function of lymphoid cells, and congenital deficiency of this enzyme is associated with severe combined immunodeficiency disease. The activity of ADA has changed in diseases characterized by the alteration of cell-mediated immunity such as rheumatoid arthritis, systemic lupus erythematosus and tuberculosis, so ADA has been considered as a nonspecific marker of cell-mediated immunity. In this study we examined changes in serum total ADA activity and the patterns of two ADA isoenzymes, ADA1 and ADA2 in normal pregnant women, and evaluated the possible role of the alteration of cell mediated immunity during normal pregnancy as causes of changes in ADA activity. We measured serum activities of total ADA, ADA1 and ADA2 in normal pregnant women in the third trimester (n=24) and age-matched healthy nonpregnant women (n=24). Peripheral blood lymphocytes and monocytes were also measured. In normal pregnant women, serum total ADA activity averaged 10.5 +/- 0.5 U/L, which was significantly lower than in nonpregnant women (14.0 +/- 0.5 U/L ) (p<0.05), and mean serum ADA2 activity also significantly reduced that of nonpregnant women (p<0.05). There was no significant difference in ADA1 activity in normal pregnant and nonpregnant women. The decrease in total ADA activity was accompanied by the decrease in lymphocyte count. These results suggest that reduced serum total ADA activity reflects decrease in ADA2 activity, and which may be in part associated with depressed cell-mediated immunity during normal pregnancy. PMID- 12592420 TI - Elastic fibres in the human placenta. AB - Recent studies described the presence of elastic fibres within stem villi of human term placenta. This particular study focuses on changes of the elastic fibre system in placental villous tree from placentas of different gestational ages. The presence of elastic fibres was researched by light and electron microscopy in first-, second-trimester and term human placentas. Light microscopic analysis revealed elastic fibres in the stroma of main stem villi and in large vessels present in the chorionic plate of term placentas. Transmission electron microscopy revealed a wide variation in microfibrillar component deposition in placentas from all the gestational ages studied. Although minimal amount of the microfibrillar component was observed only in some main stem villi of second-trimester placentas, abundant microfibrillar material was present in all types of placental villi in placental sections from term pregnancies. The amorphous component was only occasionally identified in sections from the chorionic plate of second trimester placentas. In ultra-thin sections from placental tissues from the 37th to the 42nd weeks of gestation, we can observe the amorphous component of the elastic fibres. Bundles of microfibrils with scanty participation of the amorphous component of the elastic fibres can be observed in stem villi. In mature intermediate villi, cells and their processes with distinct plasma membranes were seen close to some bundles of microfibrillar component, at times with small spots of the amorphous component. This study shows that elastic fibres of the villous stroma, are mainly composed of the microfibrillar component, while the amorphous component appears more frequently in advanced stages of villous differentiation of term placentas. PMID- 12592421 TI - A study of non-closure of the peritoneum at vaginal hysterectomy. AB - The aim of this case-controlled study was to determine whether non-closure of the peritoneum is detrimental in vaginal hysterectomy. 233 patients who underwent total vaginal hysterectomy (TVH) or laparoscopically assisted vaginal hysterectomy (LAVH) at the University of Vienna/Austria were analyzed. Cohorts of patients were formed according to their peritonealization status (open, n=117, vs closed peritoneum, n=116) and further stratified according to the type of surgical procedure: simple TVH ( n=115), TVH with concurrent vaginal repair and/or urinary incontinence surgery ( n=91) and LAVH ( n=27). No significant differences could be observed in analyzed surgical outcome (operation time, blood loss and analgesia). Complications (fever, infection, hemorrhage or revision) were similar whether the peritoneum was closed or not. After simple TVH, resumption of bowel function took place earlier in patients with open peritoneum than in those where it had been sutured (1.9 vs 2.4 days, P=0.001). No readmission for prolapse of the vaginal vault was recorded. Non-closure of the peritoneum at vaginal hysterectomy appears to be safe. Omission of peritoneal closure reduces the potential risk of injury and has a beneficial effect on bowel function. PMID- 12592422 TI - Changes in the proportion of T helper 1 and T helper 2 cells in cord blood after premature rupture of membranes. AB - This study investigated changes in the proportion of T helper (Th)1 and Th2 cells in cord blood after premature rupture of membranes (PROM), and evaluate the effects of PROM on the intrauterine fetal immune status. The proportion of CD3 positive T cells secreting interferon (IFN)-gamma as an index of Th1 cells, and interleukin (IL)-4 as an index of Th2 cells in cord blood of 12 newborns with and without PROM, were analyzed by flow cytometry. In cord blood of newborns with PROM, the proportion of IFN-gamma secreting cells significantly increased, and the proportion of IL-4 secreting cells was rather high but not significantly higher than that of newborns without PROM. These changes eventually caused a shift in the Th1/Th2 ratio to Th1 dominance in PROM. There was no significant correlation between the proportion of IFN-gamma secreting cells and the duration of PROM before the onset of labor. These results suggest that the increase in the proportion of IFN-gamma secreting cells after PROM, which eventually cause the Th1/Th2 ratios to show the Th1 predominance, may reflect in part intrauterine fetal immune responses to PROM. PMID- 12592423 TI - Alterations of thermoregulation in women with hyperemesis gravidarum. AB - We investigated the thermoregulatory alterations to coldness in hyperemesis gravidarum. In pregnant women with hyperemesis (n=22), thermographic measurements for skin temperature and heat dissipation from the back were made during immersions of hands in 15 degrees C water for 50 s. A thermogenic ratio (TR) was calculated as (A-B)/B, where A and B represent the rate of heat dissipation (Cal/hr) from the interscapular and the whole back area, respectively. Simultaneously, changes in serum nonesterified fatty acids (NEFAs) were measured in response to the stimulus. A comparison was made between the women with and without hyperemesis (n=13). Skin temperature did not respond to slight cold stimulus. However, TR increased rapidly and kept increasing (P<0.01) and NEFAs increased (P<0.05) only in subjects with fasting hyperemesis. Increase in TR was closely associated with that of NEFAs (P<0.05) and weight loss. Characteristic thermoregulatory response is found in the interscapular skin region of hyperemesis gravidarum. This may be caused by elevated lipolytic activity associated with interscapular region in undernourished pregnant women. PMID- 12592424 TI - Relation between plasma endothelin-1 levels and T helper-1:T helper-2 immunity in women with twin pregnancies. AB - The aim of this study was to investigate the relationship between plasma ET-1 levels and T helper-1 (Th1):Th2 immunity in women with twin pregnancies. The percentage of Th1 and Th2 cells and the Th1:Th2 cell ratios in peripheral blood from 13 normal singleton pregnant women and 13 patients with twin pregnancies at 29-34 weeks' gestation were calculated using flow cytometry. The plasma ET-1 was also determined using a modified radioimmunoassay. The plasma ET-1 concentrations and the cell ratios of Th1:Th2 in twin pregnancies were significantly lower than those in singleton pregnancies. A positive correlation was found between plasma ET-1 levels and the ratios of Th1:Th2 cells in the whole subjects. Our results indicate that decreased ET-1 levels are associated with Th1:Th2 immunity in twin pregnancies. PMID- 12592425 TI - Is prenatal HLA typing of uncultured amniocytes before the collection of related allogenic cord blood helpful? AB - The collection of related allogenic cord blood is gaining increasing importance in families with one child affected by haematopoietic disease. Within a family, there is only a 25% chance of a full HLA match between siblings. 50% of all collected cord blood samples cannot be used because of poor quality. Because of this, the determination of HLA type is useful for planning the collection of related allogenic cord blood transplants. We studied whether HLA typing is possible during late pregnancy if amniocentesis has not been performed during the first trimester. HLA -A, -B and -DRB loci were detected in amniotic fluid, as well as in corresponding cord blood and maternal blood using PCR-SSP. For the first time, HLA typing was performed from uncultured amniocytes. Unambiguous results were obtained from all samples. Fetal HLA-genotype in amniotic fluid was confirmed by typing results from corresponding cord blood. HLA typing of uncultured amniocytes during late pregnancy is a reliable and fast method. For the first time, prenatal HLA typing by amniocentesis after week 38 of gestation is possible in less than 8 h and without fetal risk. PMID- 12592426 TI - Breast metastasis from uterine leiomyosarcoma: a case report. AB - Breast metastasis from extra-mammary malignancy is rare, constituting 2% of breast tumors. Uterine leiomyosarcoma is a rare soft tissue neoplasm comprising 1% of all uterine malignancies. This report presents the first case of uterine leiomyosarcoma metastasis of the breast. A 44-year-old woman, G4P3AA1, presented with a one-week history of painless bilateral breast lumps. Twelve years previously she had undergone a right salpingo-oophorectomy and hysterectomy for uterine leiomyosarcoma. An excisional biopsy of the breast nodule was performed, and the histopathological study revealed the nodule was composed of spindle cells, typical of breast metastasis with uterine leiomyosarcoma found in the immunohistochemical study. The patient was healthy and symptom free during the 12 month post-operative follow-up period. PMID- 12592427 TI - Critical pulmonary stenosis with intact ventricular septum and fetal arrhythmias. AB - A 23-year-old woman, gravida 1, was referred to our hospital for possible fetal distress at 32 weeks of pregnancy. A fetal cardiotochogram showed a reactive pattern, but mild continuous bradycardia and an intermittent pulse were observed, regarded as a sinus type and a type of A-V block, respectively. The continuous deceleration of the heart rate to 95 bpm was observed frequently at 35 weeks, 3 days of gestation. Thus, an emergent cesarean section was performed and a viable 2,082 g female infant was delivered. The neonate gradually became cyanotic, and an echocardiogram was performed. The neonate was regarded as a right ventricular outflow obstruction with intact ventricular septum. Unlike other cases, the infant revealed a moderately developed right ventricle despite a severely stenotic tricuspid valve. The infant died 27 days after birth and an autopsy established the diagnosis of critical pulmonary stenosis with intact ventricular septum. Right ventricular myocardial sinusoidal-coronary artery connections, one of the major features of this type of heart anomaly, was speculated to be involved in the cause of fetal bradyarrhythmias. PMID- 12592428 TI - Treatment of choledocholithiasis in pregnancy: a case report. AB - Some difficulties have been encountered in treating pregnant patients with choledocholithiasis because of the probable harmful effect of scopic irradiation on fetus when endoscopic retrograde cholangiopancreatography (ERCP) is necessary. In this paper, we present such a patient, in whom endoscopic stone extraction without scopic examination was successfully carried out in the guidance of magnetic resonance cholangiopancreatography (MRCP) before the ERCP procedure. PMID- 12592429 TI - Postpartum ovarian vein thrombosis: an unpredictable event: two case reports and review of the literature. AB - Ovarian vein thrombosis (OVT) is a rare but serious postpartum complication that, in most cases, occurs in the right ovarian vein. Certain diagnosis, following clinical suspect because of lower quadrant tenderness and fever that alone does not respond to adequate broad-spectrum antibiotics, is now based on computed tomography (CT), although other imaging techniques, such as color Doppler ultrasonography and magnetic resonance (MR) imaging, are useful. Heparin and intravenous antibiotics are the mainstay of treatment so as to avoid laparotomy. We report on the management of two cases of postpartum OVT. PMID- 12592430 TI - Postpartum hemorrhage and emergency hysterectomy in a patient with mitochondrial myopathy: a case report. AB - Mitochondrial myopathies are a rare biochemical group of disorders of the mitochondrial respiratory chain. We report the first case in the literature of a pregnant woman with mitochondrial myopathy who, after cesarean section, had a severe and massive postpartum hemorrhage that required emergency supracervical hysterectomy. We discuss the case and review the literature. PMID- 12592431 TI - Complete bilateral crossed ureteral duplication observed during a radical hysterectomy with pelvic lymphadenectomy for ovarian cancer. A case report. AB - Complete bilateral ureteral duplication is a rare malformation. We report the case of a patient diagnosed with ovarian cancer, in which complete crossed bilateral ureteral duplication was observed during a planned radical hysterectomy with pelvic lymphadenectomy operation. If unilateral duplication is observed on one side, the existence of a similar malformation on the opposite side should always be suspected. Although bilateral ureteral duplication is an even rarer malformation than unilateral, it should be always considered as a differential diagnosis. PMID- 12592432 TI - Atypical endometrial hyperplasia in an 18-year-old woman. AB - The natural history and the factors that lead to the acquisition of atypia in endometrial hyperplasias in young aged women, especially under the age of 20, have not been fully elucidated. In such cases, although there exists a considerable risk of progression to carcinoma, a conservative antiestrogenic treatment is primarily indicated, in attempt to preserve the reproductive ability of the young woman. We report of a 18-year-old girl with atypical hyperplasia of the endometrium, a diagnosis confirmed by reviewing of the histologic material by specialized gynecopathologists. The patient has been treated with gonadotropin releasing hormone agonist (leuprolide acetate) and tibolone for 1 year, which led to endometrial atrophy and amenorrhea, without hypoestrogenic side effects. Six months after cessation of the therapy the endometrial hyperplasia relapsed (this time without atypia), but in about 2 years of follow-up and after short courses of treatment with clomiphene citrate and progestins the biopsy of the endometrium revealed a functional endometrium and the patient presents with an almost regular menstrual cycle. PMID- 12592433 TI - Intravenous leiomyomatosis. AB - INTRODUCTION: Intravenous leiomyomatosis (IVL) is rare and it is characterized by intravascular nodular masses of histologically benign smooth muscle that may extend variable distances. Although histologically benign, IVL might be malignant in its mode of behavior. CASE REPORT AND DISCUSSION: A case of IVL is reported with emphasis on immunohistochemical analysis and recent literature. PMID- 12592434 TI - Umbilical endometriosis without previous pelvic surgery: a case report. AB - A 27-year-old woman with a periodically bleeding out of the umbilicus was found to have umbilical endometriosis. She was never pregnant before and had no pelvic surgery. The umbilical lesion was excised following a diagnostic laparoscopy revealing extragenital endometriosis. Umbilical endometriosis is a very rare disease, but should be considered in the differential diagnosis of umbilical lesions. PMID- 12592435 TI - A stepwise transcervical resection of a large uterine myoma after vaginal delivery. AB - CASE REPORT: This study reported a stepwise (4-times) transcervical resection (TCR) performed on a patient with a large uterine myoma. The patient (36 years old) was at 6 weeks of pregnancy when at the first medical examination a uterine myoma, as large as the head of newborn infant, was discovered. After the delivery, myomectomy was performed by way of conserving the uterine. Since the myoma nodule was large, TCR was performed several times to completely resect it. CONCLUSION: TCR has been indicated only for small submucosal myoma; however, this study shows that TCR utilizing a stepwise technique can also be indicated for larger myoma. PMID- 12592436 TI - Condylomata gigantea in anal and perianal region: surgical and CO2 laser treatment. AB - CASE REPORT: We present a case of 28-year-old female patient with condylomata gigantea (Buschke-Lowenstein tumor) in anal and perianal region with propagation on vulva and vagina. The local surgical excision and CO(2) laser treatment were performed. Histological examination showed presence of HPV type 11 without malignant potential. RESULT: Three months later, there was no recurrence. PMID- 12592437 TI - White adipose tissue composition in the free-ranging fat-tailed dwarf lemur (Cheirogaleus medius; Primates), a tropical hibernator. AB - In temperate species, hibernation is enhanced by high levels of essential fatty acids in white adipose tissue. Essential fatty acids cannot be synthesized by mammals, thus nutritional ecology should play a key role in physiological adaptations to hibernation. Tropical hibernators are exposed to different physiological demands than hibernators in temperate regions and are expected to be subject to different constraints. The aims of this study were to assess whether or not the tropical hibernator Cheirogaleus medius shows biochemical changes in its white adipose tissue before and during hibernation. A capture recapture study was combined with feeding observations in western Madagascar. Before and after hibernation, 77 samples of white adipose tissue from 57 individuals of C. medius, as well as dietary items eaten during pre-hibernation fattening, were sampled and analyzed for their fatty acid composition. In contrast to temperate hibernators, C. medius exhibits extremely low essential fatty acid concentrations in its white adipose tissue (2.5%) prior to hibernation. The fatty acid pattern of the white adipose tissue did not change during pre-hibernation fattening and did not reflect dietary fatty acid composition. During hibernation, fat stores showed only minor but significant compositional changes. Because of its prevalence, the main fuel during hibernation was the monounsaturated oleic acid, which seemed to be preferentially synthesized from dietary carbohydrates. Results suggest that essential fatty acids do not represent an ecological limitation for hibernation in the tropics, at least not in the fat-tailed dwarf lemur. PMID- 12592438 TI - Ovarian mitochondrial cytochrome b mRNA levels increase with sexual maturity in freshwater eels (Anguilla spp.). AB - Differential display of mRNA was used to identify an upregulated gene in ovaries of artificially maturing Japanese eels (Anguilla japonica). Accordingly, mitochondrial (mt) cytochrome b, whose transcript levels increased from early to late vitellogenesis, was isolated, cloned and sequenced. Temporal trends in artificially maturing eels were compared with those in naturally and artificially maturing New Zealand eels (longfinned eel, Anguilla dieffenbachii; shortfinned eel, Anguilla australis) by Northern blot and in situ hybridization analysis to rule out any experimental artifacts. An increase in ovarian mt cytochrome b signals was seen when comparing immature and midvitellogenic longfinned eels, but not immature and early vitellogenic shortfinned eels, from the wild. Long-term captivity yielded reduced target mRNA levels, but abundance increased after hormonal induction of vitellogenesis. These results imply that the increase in mt cytochrome b mRNA levels during artificial maturation reflects natural development, although its onset appears to be brought forward during artificial maturation in the Japanese eel. It is suggested that increased mt cytochrome b mRNA levels result from both mitochondrial replication and increased transcription, and that they reflect the build-up of machinery for enhanced ATP synthesis at some stage of oogenesis and/or early zygote development. PMID- 12592439 TI - Effect of saline acclimation on body water and sodium compartmentalization in Pekin ducks (Anas platyrhynchos). AB - The compartmentalization of body fluids was measured in individual Pekin ducks ( Anas platyrhynchos) drinking freshwater and after sequential acclimation to 300 mM NaCl and 400 mM NaCl. Total body water, extracellular fluid volume, plasma volume and exchangeable sodium pool were measured using (3)H(2)O, [(14)C] polyethylene glycol, Evans Blue dye, and (22)Na dilution, respectively. Following acclimation to 300 mM NaCl, body mass decreased, but total body water and total exchangeable sodium pool were unaltered. Na and water were redistributed from the extracellular fluid (interstitial fluid) compartment into the intracellular fluid compartment. Following further acclimation to 400 mM NaCl, body mass, total body water and intracellular fluid volume decreased, but exchangeable sodium pool and extracellular fluid volume were unchanged. Our results suggested that, when Pekin ducks drink high but tolerable salinities, they maintain total body water, but redistribute Na(+) and water from interstitial fluid to the intracellular fluid compartment. When stressed beyond their ability to maintain total body water, they lose water from the intracellular fluid. PMID- 12592440 TI - Effect of saline intake on water flux and osmotic homeostasis in Pekin ducks (Anas platyrhynchos). AB - The physiological regulation of body water volume and concentration was evaluated in Pekin ducks, Anas platyrhynchos, slowly acclimated to increasingly saline drinking water (six equal 75 mM NaCl increments). Body mass, total body water (TBW), water flux, plasma osmolality (Osm(pl)), and ionic and osmoregulatory hormone concentrations were measured at the end of each increment. The salinity at which each variable deviates from its homeostatic set point was calculated by continuous two-phase linear regression. We hypothesized that, as drinking water salinity increases: (1) body water increases in concentration before it decreases in volume and (2) that regulating variables that help determine homeostatically set values (plasma hormone concentrations and water flux) deviate from values of freshwater ducks at lower drinking water salinities than the variables they regulate (Osm(pl), hematocrit, TBW). Osm(pl) was the first variable for which we could calculate a deviation from its homeostatically controlled value. It increases at much lower drinking water salinity than that at which TBW decreases, supporting our first hypothesis, but not our second hypothesis. We further hypothesized that, because the concentration of Pekin duck salt gland secretion is only slightly higher than that of their drinking water, they increase water flux (drinking) as salinity of drinking water increases, until the latter exceeds the secretion concentration and then they drink less. There was no change in water flux until it decreases when TBW decreases, 329 mM NaCl and 335 mM NaCl, respectively. The results do not support our hypothesis that Pekin ducks increase drinking as the salinity of their drinking water increases, but do indicate that, at tolerable salinities, Pekin ducks maintain body water volume while allowing body water osmolality to increase. At higher salinities, ducks decrease drinking and use body water to get rid of the excess salt. PMID- 12592441 TI - Melatonin is rhythmic in newborn seals exposed to continuous light. AB - We have investigated the effect of continuous light and darkness on plasma levels of melatonin in relation to the extremely large and active pineal gland typically found in newborn seals. Plasma levels of melatonin in captive newborn harp (Phoca groenlandica) and hooded seals (Cystophora cristata) were generally extremely high, with peak concentrations ranging from 0.8 ng/ml to 62.3 ng/ml. Moreover, plasma melatonin showed a similar, pronounced rhythmicity, both outdoors under natural light conditions (hooded seal only) and indoors under either 30 h of continuous light (490 lux) or 30 h of darkness (0 lux). In all animals, the melatonin rhythm was closely associated with the outdoor light-dark cycle. We suggest that the melatonin rhythmicity in newborn seals is mainly under circadian control and that it originates by maternal influence in the foetus. Daytime plasma concentrations of melatonin were also measured in foetal hooded seals and their mothers. The foetal melatonin level was similar to daytime levels in newborns and was about five times higher than in their mothers, which indicates a significant flow of foetal melatonin to the mother. We speculate that the large pineal gland and high melatonin levels in the newborn seals are temporary consequences of a foetal strategy to affect the maternal blood supply during diving. PMID- 12592442 TI - Standard energetics of leaf-nosed bats (Hipposideridae): its relationship to intermittent- and protracted-foraging tactics in bats and birds. AB - Basal rates of metabolism within the insectivorous genera Hipposideros and Ascelliscus, Old World leaf-nosed bats (Hipposideridae), ranged from 58% to 77% of the mammalian standard. The larger species, Hipposideros diadema and Hipposideros maggietaylori, effectively thermoregulated at ambient temperatures down to 9 degrees C, whereas two smaller species, Hipposideros galeritus and Hipposideros cervinus, occasionally permitted body temperatures to fall below 32 degrees C. The low basal rates of metabolism in hipposiderids correlated with a predatory life-style characterized by intermittent flight from a perch to capture insects, a correlation similar to that found in nonpasserine birds. Intermittent foraging bats and nonpasserines collectively had basal rates of metabolism that averaged 75% of those that pursue insects during protracted flight. However, no difference in basal rate was found between protracted- and intermittent-foraging passerines, which had basal rates 1.8- and 2.4-times those of protracted-foraging and intermittent-foraging bats and nonpasserines, respectively. Bats, swifts, and caprimulgids that enter torpor have basal rates that are 85% of those of similar species that do not enter torpor. Body mass, order affiliation, foraging mode, and propensity to enter into torpor collectively account for 97% of the variation in basal rate of metabolism in insectivorous bats and birds. Foraging style therefore appears to be a factor contributing to the diversity in endotherm energetics. Minimal thermal conductance in the genus Hipposideros ranged from 75% to 102% of the mammalian standard. Birds have minimal thermal conductances that are 75% of mammals and intermittent foragers have minimal conductances that are 78% of protracted foragers. PMID- 12592443 TI - Radiant heat affects thermoregulation and energy expenditure during rewarming from torpor. AB - The high expenditure of energy required for endogenous rewarming is one of the widely perceived disadvantages of torpor. However, recent evidence demonstrates that passive rewarming either by the increase of ambient temperature or by basking in the sun appears to be common in heterothermic birds and mammals. As it is presently unknown how radiant heat affects energy expenditure during rewarming from torpor and little is known about how it affects normothermic thermoregulation, we quantified the effects of radiant heat on body temperature and metabolic rate of the small (body mass 25 g) marsupial Sminthopsis macroura in the laboratory. Normothermic resting individuals exposed to radiant heat were able to maintain metabolic rates near basal levels (at 0.91 ml O(2) g(-1) h(-1)) and a constant body temperature down to an ambient temperature of 12 degrees C. In contrast, metabolic rates of individuals without access to radiant heat were 4.5-times higher at an ambient temperature of 12 degrees C and body temperature fell with ambient temperature. During radiant heat-assisted passive rewarming from torpor, animals did not employ shivering but appeared to maximise uptake of radiant heat. Their metabolic rate increased only 3.2-times with a 15- degrees C rise of body temperature (Q(10)=2.2), as predicted by Q(10) effects. In contrast, during active rewarming shivering was intensive and metabolic rates showed an 11.6-times increase. Although body temperature showed a similar absolute change between the beginning and the end of the rewarming process, the overall energetic cost during active rewarming was 6.3-times greater than that during passive, radiant heat-assisted rewarming. Our study demonstrates that energetic models assuming active rewarming from torpor at low ambient temperatures can substantially over-estimate energetic costs. The low energy expenditure during passive arousal provides an alternative explanation as to why daily torpor is common in sunny regions and suggests that the prevalence of torpor in low latitudes may have been under-estimated in the past. PMID- 12592444 TI - Absorption and systemic availability of two synthetic growth hormone secretogogues and transport of glucose by the proximal small intestine of anestrus dogs after administering estradiol. AB - Pharmacokinetics for one growth hormone secretogogue (NNC 26-0722), but not for another (NN703), differ between dogs in estrus or anestrus. We examined if the differences could be mimicked by administering estradiol during anestrus and if there was a relationship with rates of small intestine absorption. Pharmacokinetics for oral doses of NN703 (1.0-1.6 mg kg(-1)) did not differ among dogs in estrus, anestrus, or anestrus and given estradiol for 1 week (days 1, 3, and 6; 40 micro g kg(-1)), whereas plasma concentrations of NNC 26-0722 increased from undetectable in untreated, anestrus dogs to several hundred nanograms per milliliter in dogs given estradiol, with maximal concentrations measured 5 min after oral dosage. Estradiol treatment increased small intestinal absorption of NNC 26-0722 by 100% (P<0.05), but did not increase absorption of NN703, and caused a 64% increase in carrier-mediated glucose transport at 50 mmol l(-1) (P<0.05) due to increased densities of transporters. These findings indicate estrus and estradiol enhance absorptive functions of the dog proximal small intestine and can affect pharmacokinetics for some orally administered drugs. PMID- 12592446 TI - Reproductive energetics of the nectar-feeding bat Glossophaga soricina (Phyllostomidae). AB - Pregnancy and lactation are energetically demanding periods for female mammals. Unique amongst mammals, bats have to allocate considerable amounts of energy into their offspring because juveniles cannot be weaned until they are capable of flying at almost adult size. Similar to other bat species, female nectar-feeding bats should increase their energy intake after parturition to meet the energy demands of offspring growth. However, previous studies have shown that nectar feeding bats differ from other similar-sized bats in having a much higher metabolic rate. Therefore, I examined how nectarivorous bats respond to the energetic challenge of reproduction. In this study, the daily energy intake of pregnant and lactating Glossophaga soricina was measured during a 6-week period prior to and a 10-week period after parturition. Body mass of G. soricina increased linearly until parturition. Within the same time period, daily flight time decreased and daily energy intake remained constant. Probably, the reduced flight activity of pregnant bats compensated for the increased power requirements of flight, thus resulting in an almost constant daily energy turnover. During 35 days after parturition, neither flight time, body mass nor daily energy intake of lactating females changed significantly. On average, the daily energy intake of pregnant, lactating or non-reproducing G. soricina was not significantly different. Possibly, for unknown reasons, female G. soricina maintain a daily energy intake of a constant high level during and beyond reproduction. PMID- 12592445 TI - Differential susceptibility to Eucalyptus secondary compounds explains feeding by the common ringtail (Pseudocheirus peregrinus) and common brushtail possum (Trichosurus vulpecula). AB - The effect of two plant secondary metabolites, tannins and formylated phloroglucinol compounds (FPCs), on the intake of Eucalyptus foliage by common ringtail (Pseudocheirus peregrinus) and common brushtail possums (Trichosurus vulpecula) was studied. We manipulated the amount of tannin that was free to bind with protein by coating foliage with polyethylene glycol 4000 (PEG) and relied on natural intraspecific variation in FPC concentrations. In contrast to ringtail possums, brushtail possums showed a greater tolerance to FPCs and ate more foliage when it was coated with PEG, suggesting that tannins limited their food intake. Brushtails detected the effects of tannins through immediate oral sensations rather than through systemic effects. Ringtail possums appeared highly tolerant of foliar tannins yet susceptible to low concentrations of FPCs. We could not detect any interaction between tannins and FPCs that affected the intake of Eucalyptus foliage by either species of possum. Although ringtail and brushtail possums are widely regarded as specialist and generalist folivores, respectively, their differential susceptibility to co-occurring secondary metabolites suggests greater complexity. Each possum species appears to be a specialist in its own right, which leads to a partitioning of available foliage. Brushtails avoid tannins and ringtails avoid FPCs. PMID- 12592447 TI - Environmental and habitat management: the case of Ethiopia and Ghana. AB - This article examines the environment and habitat management experiences of Ethiopia and Ghana in the postindependence period (1960-2000). Based on extensive archival research, semistructured focused interviews of environment and habitat officers of the World Bank, the United Nations System and the International Union for the Conservation of Nature (IUCN) and personal professional field experiences, the paper argues that the uncritical adoption of externally generated discourses, narratives, policy guidelines, and strategies of environmental and habitat management has structured thought and action in both countries. The experience of both countries in defining and responding to environmental and human settlement management is explored from a political ecology perspective. The analysis indicates that both countries have essentially adopted a technocratic, state-centered, and unsustainable management strategy framework based on population control, poverty reduction, sustainable development, and capacity-building. It also suggests that international organizations such as the World Bank, INCN, and the United Nations system have been important sources of thought and action in both countries. Conversely, regional international organizations such as the Economic Commission for Africa, the Organization of African Unity and the African Development Bank have largely served as conduits for the diffusion of global discourses, narratives, policies and strategies. The need for adopting management policies and strategies that are based on principles of multiple engagement, decentralization, incentives, public education, and participation is underscored. PMID- 12592448 TI - Sediment flux to the sea as influenced by changing human activities and precipitation: example of the Yellow River, China. AB - Since the 1970s, the sediment flux of the Yellow River to the sea has shown a marked tendency to decrease, which is unfavorable for wetland protection and oil extraction in the Yellow River delta. Thus, an effort has been made to elucidate the relation between the sediment flux to the sea and the drainage basin factors including climate and human activities. The results show that the sediment flux to the sea responds to the changed precipitation in different ways for different runoff and sediment source areas in the drainage basin. If other factors are assumed to be constant, when the annual precipitation in the area between Longmen and Sanmenxia decreases by 10 mm, the sediment flux to the sea will decrease by 27.5 million t/yr; when the precipitation in the area between Hekouzhen and Longmen decreases by 10 mm, the sediment flux to the sea will decrease by 14.3 million t/yr; when the precipitation in the area above Lanzhou decreases by 10 mm, the sediment flux to the sea will decrease by 17.4 million t/yr. A multiple regression equation has been established between the sediment flux to the sea and the influencing factors, such as the area of land terracing and tree and grass planting, the area of the land created by the sediment trapped by check dams, the annual precipitation, and the annual quantity of water diversion by man. The equation may be used to estimate the change in the sediment flux to the sea when the influencing variables are further changed, to provide useful knowledge for the environmental planning of the Yellow River drainage basin and its delta. PMID- 12592449 TI - Compliance with regulations by "swim-with-dolphins" operations in Port Phillip Bay, Victoria, Australia. AB - Managing the activities of commercial wildlife viewing tends to involve either restricting the number of industry participants and/or regulating the activities or industry participants. We report on operator compliance with regulations regarding humans swimming with free-ranging bottlenose dolphins ( Tursiops sp.) in Port Phillip Bay, Victoria, Australia. A total of 128 commercial dolphin-swim trips was studied between September 1998 and April 2000. Four permit conditions were investigated: approach type, swim time, time in proximity of dolphins, and presence of "fetal fold" calves. Results demonstrate noncompliance by operators to all of the four permit conditions studied. Compliance with temporal conditions was poorer than with other conditions. When conducting studies on the extent to which tourism affects cetaceans, investigators should consider whether tourist operations comply with existing regulations or guidelines. PMID- 12592450 TI - Population, development, and waste management in Botswana: conceptual and policy implications for climate change. AB - Based on government and other relevant documentation, this paper explores the conceptual linkage between population, development, and waste management in Botswana and the implications of this relationship for global climate change. Population is increasing, albeit at a decreasing rate. Spatially, the population is becoming more and more concentrated as the rates and level of urbanization increase. Economic growth has remained consistently high. The combined effect of population dynamics and economic development are having a noticeable imprint on the environment in the form of increased waste generation. Poor waste management poses a real threat to environmental sustainability in general and climate change in particular because of inadequate technology, weak institutional mechanisms to enforce regulations, and low levels of sensitization among the public to deal with the problem. Mitigation measures are suggested to minimize the negative effects of waste management on climate change. PMID- 12592451 TI - The tribal environment: solid waste in Indian country. AB - The issue of solid waste management in Indian country is multidimensional in scope because it affects more than just regulatory concerns. There are more than 550 federally recognized Alaska Native and American Indian Tribes in the United States. Tribes are sovereign nations that have a special relationship to the federal government and a unique legal status. The environmental problems faced by tribes are many, and it is only fair that tribes, as sovereigns, specify the levels of protection on their lands. The one-size-fits-all regulatory approach to environmental problems and solid waste management in particular does not work and often leads to conflict between tribes and the federal and state governments. Inherent tensions also exist between tribes and various levels of government concerning jurisdiction of lands and managing solid waste. These intergovernmental relationships are often complex and present unique challenges to all. More research needs to be done on targeting resources to meet the capacity-building needs of tribes, as well as the overall environmental management needs of Indian country under the federal trust obligation. Successful intergovernmental relationships can be fostered through partnership arrangements between tribes and federal, state, and local governments. In the area of solid waste such partnerships have worked. It requires that all levels of government deal with tribes with careful consideration of their cultural, historic, and socioeconomic aspects, which are often intertwined. PMID- 12592452 TI - Examining landscape dynamics at a watershed scale using Landsat TM imagery for detection of wintering hooded crane decline in Yashiro, Japan. AB - It is usually inappropriate to define rectangular land areas or administrative units as the extent for quantifying landscapes that possess hierarchical structure. As a functional unit established by geophysical relationships, the watershed is one of many natural scales in the hierarchical landscape. We examined the dynamics of the Yashiro watershed of Japan at the landscape level using pattern metrics based on Landsat thematic mapper (TM) imagery from 1985 to 1998. This watershed provides important habitats for the hooded crane (Grus monachus), a vulnerable species. While its world population has remained stable, the number wintering at Yashiro has declined in recent years. Changes in landscape metrics reveal that the spatial pattern within the watershed underwent homogenization due to depopulation of local people and shifts in local energy requirements and forest management policy at Yashiro. Specific changes include: a decrease in bare land area from 6.2% to 1.0% of the landscape, increased forest cover from 69.2% to 76.1%, reduction in patch number from 1194 to 616 and enlarged mean patch size, and a decrease in total edge from 223,740 m to 158,040 m. The rate of change in landscape metrics indicates a rapid change towards homogeneity in the landscape since 1990. The temporal changes in hooded crane populations corresponded to the changes in landscape. An alternative explanation has been proposed that decline of the species is influenced by landscape dynamics affecting both habitat selection and food resources. Conservation at the watershed scale is suggested to be complementary to the current conservation measures of the species. PMID- 12592453 TI - Effects of mechanical fracturing and experimental trampling on Hawaiian corals. AB - In situ trampling occurred under experimental conditions to quantify the differences in the responses to anthropogenic trampling in four dominant species of Hawaiian corals, Porites compressa, Porites lobata, Montipora capitata, and Pocillopora meandrina. Trampling was simulated daily for a period of nine days at which time further breakage was minimal. Forty treatment colonies produced 559 fragments. Trampling was followed by an 11-month recovery period. Coral colony and fragment mortality was low. All four species were highly tolerant of inflicted damage, suggesting that some species of corals can withstand limited pulse events that allow time for recovery. Growth rates following trampling were significantly lower in the treatment groups for three of the four species. This study demonstrated that very few trampling events can produce significant changes in growth even after a long recovery period. Survivorship of fragments is clearly size- and species-dependent in M. capitata and P. compressa. Smaller fragments (<5 cm) had higher mortality than larger fragments (>5 cm). High breakage rates for M. capitata and P. compressa are consistent with the near shore, low-energy regions they inhabit-the same environment frequented by skin divers and waders. Mechanical tests were conducted to determine tensile and compressive strengths. Pocillopora meandrina exhibited the strongest skeletal strength, followed in decreasing order by Porites lobata, Porites compressa, and Montipora capitata. The skeletal strength obtained from the experiments correlate with the wave energy present in the environments in the regions they inhabit, suggesting that structural strength of corals is an adaptive response to hydraulic stress. PMID- 12592454 TI - A geomorphic explanation for a meander cutoff following channel relocation of a coarse-bedded river. AB - The Veteran's Fishing section of the Blackledge River in central Connecticut was relocated in the late 1950s. The relocation resulted in an unstable channel despite extensive efforts to prevent erosion. Overbank erosion and meander cutoffs were investigated using detailed survey data, characterizations of sediment deposits, flow modeling, and a moment-stability analysis. Limited reworking of revetment boulders indicate that riprap bank material was immobile during a 1979 flood event responsible for the formation of the cutoff channel. A moment-stability analysis factor-of-safety value of 1.1 supports the conclusion that riprap was not directly eroded from the banks. Alluvial particles with d(95) values ranging up to 120 mm were deposited along a bar downstream from the cutoff channel at flows estimated to be below a 1.5-year recurrence interval flow. Development of the bar deposit resulted in locally elevated water surfaces at high flow. The resulting overbank flow across the meander neck to the adjacent downstream bend led to the creation of an upstream migrating knickpoint, the erosion of approximately 16,000-year-old sediments, and the subsequent meander cutoff. The results of the study indicate that traditional erosion-control measures cannot prevent extreme channel adjustments if the geomorphic processes that control sediment continuity also are not considered. PMID- 12592455 TI - Radiative forcing due to anthropogenic greenhouse gas emissions from Finland: methods for estimating forcing of a country or an activity. AB - The objective of this study was to assess the radiative forcing due to Finnish anthropogenic greenhouse gas emissions in three scenarios. All the Kyoto Protocol gases, i.e., CO(2), CH(4), N(2)O, and fluorinated gases, were included. The calculations showed that forcing due to Finnish emissions will increase in the case of all gases except methane by the year 2100. In 1990, radiative forcing due to Finland's emission history of all Kyoto Protocol gases was 3.2 mW/m(2), of which 71% was due to carbon dioxide, 17% to methane, and the rest to nitrous oxide. In 1990 the share of fluorinated gases was negligible. The share of methane in radiative forcing is decreasing, whereas the shares of carbon dioxide and of fluorinated gases are increasing and that of nitrous oxide remains nearly constant. The nonlinear features concerning additional concentrations in the atmosphere and radiative forcing due to emissions caused by a single country or activity are also considered. Radiative forcing due to Finnish emissions was assessed with two different approaches, the marginal forcing approach and the averaged forcing approach. The impact of the so-called background scenario, i.e., the scenario for concentration caused by global emissions, was also estimated. The difference between different forcing models at its highest was 40%, and the averaged forcing approach appeared to be the more recommendable. The effect of background concentrations in the studied cases was up to 11%. Hence, the choice of forcing model and background scenario should be given particular attention. PMID- 12592456 TI - Alteration in rhizosphere soil properties of afforested Rhamnus lycioides seedlings in short-term response to mycorrhizal inoculation with Glomus intraradices and organic amendment. AB - The reestablishment of autochthonous plant species is an essential strategy for recovering degraded areas under semiarid conditions. A field experiment was carried out to assess the short-term effect of two reafforestation methods involving mycorrhizal inoculation and compost addition on soil quality parameters and Rhamnus lycioides seedling growth. The nutrient content (NPK) and enzymatic activities (dehydrogenase, urease, protease-BAA, acid phosphatase and beta glucosidase) increased and bulk density decreased in the rhizosphere soil with the organic amendment. Biomass C of rhizosphere soil increased by at least 240% with respect to the control soil after mycorrhizal inoculation and the combination of compost addition + mycorrhizal inoculation. Both mycorrhizal inoculation and composted organic residue addition increased R. lycioides seedling growth in the same proportion. In the short term, we conclude that the application of both reafforestation methods not only enhances the establishment of R. lycioides seedlings, but also improves soil quality. PMID- 12592457 TI - Capacity to influence sources of heavy metals to wastewater treatment sludge. AB - Wastewater treatment sludge contains valuable nutrients, yet represents a residue problem due to the amount of heavy metals, which is one of the factors preventing the use of sludge as fertilizer. In Sweden, the government has a declared aim of closing the ecocycles of nutrients to arable land, while at the same time the existing limit values are among the lowest in the world. This paper focuses on the opportunities for Swedish wastewater utilities (WWUs) to manage and reduce sources of heavy metals in order to get sludge approved and recycled. A case study focusing on sources of the heavy metals Cd, Cu, Hg, Ni, Pb, and Zn and the WWUs ability to influence the source by means of existing regulations was traced and categorized. A substantial fraction of the heavy metals were shown to be caused by diffuse emissions, such as the use of goods and products in society. For most sources discussed in the paper, a WWU can only manage the sources to a limited extent, if at all. This study shows that the WWUs cannot manage both recycling of nutrients and a decrease in heavy metals ending up in sludge. Diffuse emissions of heavy metals must be examined in context, not as a separate phenomenon or an issue for a single actor. The existing environmental legislation does not handle diffuse sources, yet long-term strategies for reducing the emissions to wastewater treatment must involve the reduction of diffuse emissions. PMID- 12592458 TI - Remotely sensed data for ecosystem analyses: combining hierarchy theory and scene models. AB - Remotely sensed data have been used extensively for environmental monitoring and modeling at a number of spatial scales; however, a limited range of satellite imaging systems often constrained the scales of these analyses. A wider variety of data sets is now available, allowing image data to be selected to match the scale of environmental structure(s) or process(es) being examined. A framework is presented for use by environmental scientists and managers, enabling their spatial data collection needs to be linked to a suitable form of remotely sensed data. A six-step approach is used, combining image spatial analysis and scaling tools, within the context of hierarchy theory. The main steps involved are: (1) identification of information requirements for the monitoring or management problem; (2) development of ideal image dimensions (scene model), (3) exploratory analysis of existing remotely sensed data using scaling techniques, (4) selection and evaluation of suitable remotely sensed data based on the scene model, (5) selection of suitable spatial analytic techniques to meet information requirements, and (6) cost-benefit analysis. Results from a case study show that the framework provided an objective mechanism to identify relevant aspects of the monitoring problem and environmental characteristics for selecting remotely sensed data and analysis techniques. PMID- 12592459 TI - Sustainable management for the eastern Mediterranean coast of Turkey. AB - The objective of this article is to propose a program for the integrated coastal zone management that is required to stimulate and guide sustainable development of the Mediterranean coastal zone of Turkey. Improved data collection, quality control, analysis, and data management will provide a firm basis for future scientific understanding of the East Mediterranean coast of Turkey and will support long-term management. Various innovative procedures were proposed for a promising ecosystem-based approach to manage coastal wetlands in the Mediterranean: remote data acquisition with new technologies; environmental quality monitoring program that will provide a baseline for monitoring; linking a Geographic Information System (GIS) with natural resource management decision routines in the context of operational wetlands, fisheries, tourism management system; environmental sensitivity analysis to ensure that permitted developments are environmentally sustainable; and use of natural species to restore the wetlands and coastal dunes and sustain the system processes. The proposed management scheme will benefit the scientific community in the Mediterranean and the management/planning community in Eastern Turkey. PMID- 12592460 TI - Advances in urogenital radiology. PMID- 12592461 TI - Evaluation of retroperitoneal and pelvic lymph node metastases with MRI and MR lymphangiography. AB - Local, regional lymph node involvement is an essential prognostic factor and an important determinant of treatment choices for patients with retroperitoneal and pelvic cancer. Current cross-sectional imaging modalities, including computed tomography and magnetic resonance (MR) imaging, use the nonspecific criterion of size and are limited in their ability to differentiate benign from malignant lymph nodes. MR lymphography is a promising imaging modality in differentiating benign from metastatic lymph nodes and provides information on lymph node morphology and function. Ultrasmall superparamagnetic iron oxide (USPIO) particles with a long plasma circulation time are suitable as an MR contrast agent for intravenous MR lymphography. They are taken up by macrophages in normally functioning nodes and reduce the signal intensity of tissue in which they accumulate because of T2 and susceptibility effects of iron oxide. In metastatic nodes, macrophages are replaced by cancer cells, which lack reticuloendothelial activity and cannot take up USPIO. The main mechanisms that might explain a heterogeneous node appearance after USPIO injection are discussed. In published reports, USPIO has shown high degrees of sensitivity and specificity for characterizing lymph nodes in cancer patients. We review the development of USPIO compounds, their imaging characteristics, and our clinical experience. PMID- 12592462 TI - Functional MRI of the kidney. AB - Functional MR imaging of the kidney has a great potential of development because the functional parameters, which can be approached noninvasively, are multiple: glomerular filtration, tubular concentration and transit, blood volume and perfusion, diffusion, and oxygenation. Until now, its limitations in clinical applications are due to the difficulties in obtaining reproducible and reliable information in this mobile organ and, sometimes, in understanding the physiologic substrate of the signal changes observed. These approaches require either endogeneous contrast agents, such as water protons (for perfusion and diffusion) or deoxyhemogobin (for oxgenation), or exogeneous contrast agents such as gadolinium chelates (for filtration and perfusion) or iron oxide particles (for perfusion). Clinical validation of these methods and evaluation of their clinical impact are now worthwhile before diffusing them in clinical practice. PMID- 12592463 TI - Functional renal imaging with nuclear medicine. PMID- 12592464 TI - New imaging of the anterior male urethra. AB - Retrograde urethrography is the method commonly employed in the detection of anterior male urethra pathology. Ultrasonography and magnetic resonance imaging have been proposed recently as new methods that can replace conventional radiography. Normal urethral anatomy can be easily studied after saline distention of the urethra, which creates high contrast relative to adjacent structures such as mucosa and corpus spongiosum. Sonourethrography with high frequency probes has a high spatial resolution and has proved to be sensitive, specific, and accurate in the diagnosis of anterior urethral pathologies such as strictures, syringocele, tumors, and trauma. It has a major role in the selection of therapy in patients with bulbar strictures. Magnetic resonance imaging with a surface coil with a small field of view can detect penile and bulbar stenoses and tumor extension. After contrast administration, the activity of the inflammatory process in the spongiosum can be assessed and used to modify the urethroplasty technique. The clinical use of the new imaging modalities in the male anterior urethra is limited despite their demonstrated accuracy in diagnosis and presurgical staging of different diseases. PMID- 12592465 TI - European Society of Urogenital Radiology guidelines on administering contrast media. AB - The European Society of Urogenital Radiology is committed to improving knowledge and understanding about contrast media. There seemed to be little agreement about the best approach for the prevention of contrast medium-induced nephrotoxicity or generalized adverse reactions and the use of intravascular contrast media in diabetic patients receiving metformin. There was also concern that there might be discrepancies between current practice and published recommendations on these subjects. The Contrast Media Safety Committee of the European Society of Urogenital Radiology has produced practical guidelines on these important topics based on wide consensus and extensive review of the literature. The details of the guidelines are presented in this review. PMID- 12592466 TI - MR urography today. AB - Magnetic resonance (MR) urography is performed by pursuing two different imaging strategies. On the one hand, heavily T2-weighted turbo spin-echo sequences are employed for obtaining unenhanced static-water images of the urinary tract. On the other, the T(1)-weighted MR urographic technique imitates conventional intravenous pyelography and is, therefore, referred to as excretory MR urography. For this reason, a gadolinium contrast agent is injected intravenously and, after its renal excretion, the gadolinium-enhanced urine is imaged with fast T1 weighted gradient-echo sequences. Both MR urographic techniques can be combined for a comprehensive examination of the upper urinary tract. This article reviews the current technical principles, imaging capabilities, and clinical applications of T2- and T1-weighted MR urography in adult and pediatric patients. PMID- 12592467 TI - Nonobstructive renal causes of flank pain: findings on noncontrast helical CT (CT KUB). PMID- 12592468 TI - Magnetic resonance imaging of the pelvic floor. AB - Magnetic resonance imaging is a novel noninvasive imaging modality for the assessment of pelvic floor dysfunction. It relies on static sequences with a high spatial resolution to study muscle morphology (levator ani) and fast imaging dynamic sequences during contraction, rest, and straining. Prolapse of the various pelvic compartments is detected with respect to organ position relative to the pubococcygeal line during dynamic phases. Compared with clinical examination, its input appears to be especially invaluable in the posterior compartments (peritoneal and digestive) and to assess complex prolapses involving more than one pelvic compartment. It is also useful for understanding postsurgical recurrences. PMID- 12592469 TI - Outcome of intravenous urography in the year 2000. AB - BACKGROUND: We studied the referrals for intravenous urography and their outcome during the year 2000 in a university department before the switch to unenhanced computed tomography (CT) and CT urography. METHODS: From the Radiology Information System, we obtained information about age, sex, referring physician, indication, and diagnosis. RESULTS: A total of 1229 intravenous urographies was performed in 1164 patients. In patients younger than 40 years, females were examined more frequently than males, and vice versa. Forty-five percent of all patients were referred with renal colic, and 41% were referred with hematuria. Renal colic was a more frequent indication than hematuria in patients younger than 61 years, whereas the opposite was the case in patients older than 60 years. Of the 559 urographies performed due to renal colic, a calculus in the upper urinary tract was found in 27% and a change indicating a tumor in 4%; the incidence of calculus increased with age up to 80 years. Of the 487 urographies performed due to hematuria, a calculus in the upper urinary tract was found in 6% and a change indicating a tumor in 15%. In the 277 patients younger than 40 years, an abnormality was diagnosed in 25% and a potential tumor was diagnosed in only 1.4%. The latter could not be confirmed at other examinations. Of the 887 patients older than 40 years, an abnormality was found in 45% and a potential tumor in 11%. CONCLUSION: A switch from urography to unenhanced CT and CT urography should not be done without an audit of the referrals and their outcomes. Patients with hematuria and younger than 40 years seem more appropriate for ultrasound than for CT because the incidence of tumors and calculus disease is low. No transitional cellular cancer was found in these patients. PMID- 12592471 TI - Left renal vein aneurysm: imaging findings. AB - A renal vein aneurysm is a rare vascular disease. To our knowledge, only six cases have been reported. We describe a 57-year-old woman with a left renal vein aneurysm diagnosed by a combination of ultrasonography, color Doppler ultrasonography, contrast-enhanced computed tomography, and magnetic resonance imaging. PMID- 12592470 TI - Influence of age on intrarenal resistive index measurement in normal subjects. AB - BACKGROUND: We investigated the influence of age on intrarenal arterial resistive index (RI) measurement in 135 normal subjects (71 male, 64 female; age range = 17 68 years, median age = 37 years). METHODS: Each subject underwent color Doppler measurement of the intrarenal RI from three distinct interlobar arteries in the superior, middle, and inferior parts of both kidneys. The mean of six RI values obtained from both kidneys was used for analysis. The correlation of RI with different variables was investigated by linear regression and stepwise multiple linear regression. Variables analyzed were age, systolic blood pressure, diastolic blood pressure, mean blood pressure, pulse pressure, and pulse rate. RESULTS: The results of linear regression showed that age had a significantly positive correlation ( r = 0.276, p = 0.0012) and diastolic blood pressure had a significantly negative correlation ( r = -0.186, p = 0.0311) with the RI. The results of stepwise multiple linear regression showed that the combination of age and diastolic blood pressure could explain approximately 15% of the RI changes ( r(2) = 0.1535). CONCLUSION: Although there is a statistically significant positive correlation between intrarenal RI and age, the correlation is weak. This suggests that the influence of age on RI measurement is small and may be of no clinical importance. PMID- 12592472 TI - Imaging after endovascular repair of abdominal aortic aneurysm. AB - Endovascular repair of abdominal aortic aneurysm is a less invasive alternative to open surgery. With the recognition of this new treatment, however, many complications, some of them life-threatening, have been reported. Short-term and mid-term results have shown that this technology is advancing and needs close follow-up. Imaging plays a major role in the evaluation of the aorta after endolumenal repair. This article reviews the roles of different imaging techniques. PMID- 12592473 TI - Videofluoroscopic evaluation of HIV/AIDS patients with swallowing dysfunction. AB - BACKGROUND: We investigated the association of abnormalities of the swallowing mechanism in the oral and pharyngeal phases of swallowing with symptoms of dysphagia and painful swallowing in patients with human immunodeficiency virus (HIV). METHODS: Seventeen patients, two with HIV and 15 with acquired immunodeficiency syndrome (AIDS), with symptoms of dysphagia or pain on swallowing for routine barium esophagograms were studied prospectively by videofluoroscopic examination of the oropharynx in addition to a routine biphasic esophagogram. The videofluoroscopic studies were performed in conjunction with a speech pathologist. RESULTS: All 17 patients demonstrated abnormalities in the oral, pharyngeal, or esophageal phase of swallowing. Eight patients aspirated, six of whom did not exhibit a cough reflex and were classified as silent aspirators. Seven of the eight patients who aspirated had chest radiographs consistent with aspiration pneumonia. CONCLUSION: A significant number of HIV positive and AIDS patients with dysphagia or pain on swallowing have dysfunction of the swallowing mechanism and are at risk for aspiration. PMID- 12592474 TI - Gastric ulcer penetrating the anterior abdominal wall: ultrasound diagnosis. AB - We report two rare cases of penetration of the anterior abdominal wall by gastric peptic ulcers. The full diagnosis was made by ultrasound, which showed similar findings: sharply delineated, hyperechoic craters clearly traversing the gastric wall and covered by hypoechoic inflammatory components of the abdominal wall. Because ultrasound is increasingly used as a primary procedure for evaluation of abdominal complaints, efforts should be directed toward exploring the accessible portions of the gastric and duodenal walls to detect peptic ulcer disease and its complications. PMID- 12592475 TI - Visualizing the gastric wall with a 30-MHz ultrasonic miniprobe: ex vivo imaging of normal gastric sites and sites of early gastric cancer. AB - BACKGROUND: This study was performed to determine the echo layer structures of the normal gastric wall and early gastric cancer when visualized with a 30-MHz ultrasonic miniprobe. METHODS: Twelve surgically resected gastric specimens were used for an ex vivo study. Eighteen normal sites and 12 early gastric cancer sites were scanned with an Olympus (XUM-S30-25R) probe with a frequency of 30 MHz. Endoscopic ultrasound images were compared with corresponding histopathologic sections stained with hematoxylin and eosin. RESULTS: The normal mucosa was visualized as at least four alternating echo layers; the muscularis mucosa was delineated at all normal sites. Lymphoid aggregates within the mucosa could be seen. The submucosa was clearly visualized in most cases, but the muscularis propria and subserosa were seldom depicted due to attenuation of ultrasound waves. At the sites of gastric cancer, the layered architecture of the mucosa was disturbed by an irregular hypoechoic lesion. Minimal submucosal infiltration (400 and 750 micrometer) was clearly depicted in two cases, without ulceration at or around the tumor site. However, attenuation at the site of a deep ulcer scar prevented adequate visualization of the tumor extent in two other cases with ulceration. CONCLUSION: A 30-MHz ultrasonic miniprobe may provide additional imaging information of the gastric wall and could play a role in the assessment of early cancer lesions. PMID- 12592476 TI - CT of small bowel obstruction in adults. AB - The increasing use of computed tomography in evaluating patients with acute abdominal pain has revolutionized the diagnosis of small bowel obstruction in adults. Computed tomography is incontestably the most useful and powerful tool to make positive, topographic, and etiologic diagnoses of small bowel obstruction. Good knowledge of some key signs and rigorous analysis of computed tomographic images by radiologists should lead to improved diagnosis and appropriate treatment. PMID- 12592477 TI - Helical CT of large bowel obstruction. PMID- 12592478 TI - Resolving appendicitis: role of CT. AB - BACKGROUND: We assessed the role of computed tomography (CT) in the resolution of appendicitis. METHODS: This was a retrospective study over 2.5 years involving 155 patients and 172 CT scans for suspected appendicitis in an emergency setting. RESULTS: Sixty-nine studies were positive for appendicitis by CT criteria. Of these, 53 underwent surgery: 48 were positive for appendicitis, four had other pathologies (two with colon cancer, one with colitis, one with infectious enteritis), and one had a normal appendix that was removed. Three of the 69 patients were released or left the emergency room against medical advice. Twelve patients demonstrated acute appendicitis by CT criteria but were treated conservatively because their clinical conditions were stable; four of those patients had follow-up CT that showed resolution of findings. Six patients were followed clinically, without recurrence of symptoms, and two were lost to follow up. CONCLUSION: Resolving appendicitis is an entity that should be considered in a patient who has signs positive for appendicitis on CT but is doing well clinically. CT positive for appendicitis does not necessarily indicate surgery, and the decision for operation should depend on clinical and radiologic features. PMID- 12592479 TI - Granulomatous appendicitis presenting as right lower quadrant pain: CT findings. AB - BACKGROUND: We present the computed tomographic (CT) findings of granulomatous appendicitis. METHODS: Five of 652 (0.9%) patients who had undergone appendectomy for clinically suspected acute appendicitis over a 19-month period proved to have granulomatous appendicitis. One patient had surgery based on a clinical diagnosis of acute appendicitis. Four patients (three men and one woman; age range = 14-39 years) underwent abdominal CT. The CT findings were retrospectively reviewed with special attention to the appendiceal abnormalities. RESULTS: All four patients presented with subacute clinical presentation. Enlarged appendices of 4.5 and 2 cm in diameter with thickened walls of soft tissue density were found in two patients, and periappendicular inflammatory masses were found in the other two. Enlarged mesenteric lymph nodes and right lower quadrant fat stranding was seen in all four patients. Histopathology showed numerous granulomas within the inflamed appendix. CONCLUSION: Radiologists should be familiar with the rare entity of granulomatous appendicitis in patients examined by CT for suspected acute appendicitis. An insidious clinical presentation with CT findings of an exceptionally large appendix and associated periappendiceal inflammatory changes should raise the possibility of granulomatous appendicitis or carcinoma or lymphoma of the appendix. PMID- 12592480 TI - Late recurrence of malignant melanoma presenting as peritoneal "carcinomatosis". AB - Malignant melanoma is a relatively rare malignancy that arises from melanocytes and accounts for approximately 1% of all malignancies reported in the United States. Malignant melanoma can develop in any part of the skin or mucosal membranes. It metastasizes to all organs of the body and often demonstrates unpredictable metastatic behavior. Late recurrence of malignant melanoma, defined as occurring 10 or more years after diagnosis and treatment, is a rare but characteristic metastatic behavior of malignant melanoma. We present a case of a late recurrence of malignant melanoma presenting with diffuse peritoneal studding. PMID- 12592481 TI - Portacaval shunt creation using the percutaneous transhepatic-transjugular technique. AB - BACKGROUND: The feasibility and efficacy of portacaval shunt creation through the transhepatic and transjugular approach were evaluated in patients with portal hypertension whose hepatic venous anatomies were unsuitable for the standard shunt procedure. METHODS: Portacaval shunts were created via percutaneous transhepatic access in six patients. We used snares to target the portal vein and the inferior vena cava. Then a needle was introduced percutaneously and advanced through the snares. A guidewire was advanced through the needle and snared in the inferior vena cava. The wire was then withdrawn through the transjugular sheath. The rest of the procedure was completed in standard fashion. The transhepatic tracts were embolized. RESULTS: The portacaval shunt procedures were successful in all patients. There were no major complications during the procedures, but one patient died of sepsis 1 week later. Two patients developed shunt occlusion, and in one case the shunt was revised successfully. The other patient with occlusion underwent orthotopic liver transplantation. The shunts in the remaining three patients and the revised shunt were patent at 5 to 20 months of follow-up. CONCLUSION: Direct portacaval shunt creation using the percutaneous transhepatic and transjugular technique is a good alternative when standard portosystemic shunt creation is difficult or impossible. PMID- 12592485 TI - The reversibility of reduced cortical vein compliance in normal-pressure hydrocephalus following shunt insertion. AB - Superficial cortical venous compression secondary to alterations in craniospinal compliance is implicated in the pathogenesis of normal pressure hydrocephalus (NPH). A reduction in the pulsation in the outflow of the cortical veins would be expected to occur following compression of these veins and this has been shown in NPH. If cortical vein compression is a causative factor in NPH, it would be expected that cortical vein compliance as measured by pulsatility would be significantly altered by a curative procedure i.e. shunt tube insertion. My purpose is to compare the blood flow pulsatility characteristics in a group of patients with NPH before and after shunt tube insertion. I initially studied 18 subjects without pathology with MRI flow quantification studies of the cerebral arteries and veins to define the range of normality. The main study involved 18 patients with idiopathic dementia and mild leukoaraiosis who served as controls and seven patients with NPH studied before and after shunt insertion. Arterial, superior sagittal and straight sinus pulsatility was not significantly different between the patients with idiopathic dementia and those NPH patients before or after shunting. Cortical vein pulsatility before shunting in the patients with NPH was 43% lower than in those with idiopathic dementia ( P=0.006). Following shunting, cortical vein pulsatility increased by 186% ( P=0.007). There is thus reduced compliance in cortical veins in NPH which is significantly increased in patients who respond to insertion of a shunt tube. These findings suggest that reversible elevation in cortical vein pressure and reversal of the normal absorption pathway for cerebrospinal fluid may be behind the pathophysiology of NPH. PMID- 12592486 TI - Extracranial internal carotid and vertebral artery dissections: angiographic spectrum, course and prognosis. AB - We reviewed the clinical and radiological findings of 93 consecutive patients with 111 extracranial internal carotid (ICAD) and vertebral artery (VAD) dissections and one concomitant intracranial VAD; 83% of the patients had unilateral and 17% multiple vessel dissections. The diagnosis was made by intra arterial digital subtraction angiography in 92 patients and MR angiography in one. Follow-up angiography was performed in 77 cases (83%): of 49 initially stenotic arteries, 40 became completely or almost completely normal, while three showed slight improvement. Of 30 initially occluded arteries, nine had completely or partly recanalised. Of 12 pseudoaneurysms eight were unchanged at follow-up. The proximal vertebral artery was involved as often as the C1-C2 level. Recurrences were rare: a new dissection in another vessel was found in three patients. Kinking or coiling was found in 23% of the dissected internal carotid arteries. PMID- 12592487 TI - Rim enhancement of meningiomas on fast FLAIR imaging. AB - We investigated the enhancement patterns of meningiomas on fast fluid-attenuated inversion-recovery (FLAIR) images and related them to the size and histology of the tumour and the associated oedema. We studied 30 meningiomas with T2-weighted fast spin-echo (SE) images plus T1-weighted SE images with magnetisation-transfer saturation and fast FLAIR before and after contrast enhancement at 0.5 tesla. There were 21 meningiomas (70%) which showed peripheral (rim) enhancement on fast FLAIR, while only one, which showed heavy central calcification, enhanced peripherally on the SE images. Of the meningiomas with capsular enhancement on fast FLAIR 20 (95%) were more than 2 cm in diameter. The nine 9 smaller meningiomas enhanced homogeneously. This difference was statistically significant pattern ( P<0.01). All meningiomas which had associated oedema showed the capsular pattern although their number (6; 20%) was to small to analyse statistically. Only 11 (36%) tumours were examined histologically; peripheral enhancement was observed in all types of meningioma. This pattern may help to differentiate meningiomas from other extra-axial masses. PMID- 12592488 TI - MRI in a case of muslin-induced granuloma. AB - When intracranial aneurysms are deemed nonclippable, an accepted alternative neurosurgical treatment is to reinforce the aneurysm wall by wrapping or coating it with various materials, including muslin (cotton gauze). Granulomatous or "foreign-body" reactions, sometimes referred to as "muslinomas" or "gauzomas," and adhesive arachnoiditis are known but rare complications. Experience with MRI in these cases is limited. We describe the clinical and MRI features of a muslinoma developing after treatment of an anterior communicating artery aneurysm. PMID- 12592489 TI - Simultaneous involvement of third and sixth cranial nerve in a patient with Lyme disease. AB - We report a 57-year-old woman with neuroborreliosis presenting with headache, shoulder muscle pain and double vision. MRI demonstrated enhancement of the right third and sixth cranial nerves. A 3D MP-RAGE sequence was used to perform multiplanar reformations to show this more graphically. The patient was free of symptoms 1 month after completion of therapy, when thickening and contrast enhancement of the nerves were less pronounced. PMID- 12592490 TI - Diffuse dural enhancement in cavernous sinus dural arteriovenous fistula. AB - We describe unusually extensive contrast enhancement of the dura mater on MRI in a case of dural arteriovenous fistula of the cavernous sinus. PMID- 12592491 TI - Diffusion-weighted MRI of the cervical spinal cord using a single-shot fast spin echo technique: findings in normal subjects and in myelomalacia. AB - We have implemented a new diffusion-weighted MRI (DWI) sequence based on the single-shot fast spin-echo technique. We hypothesised that this would add information to conventional MRI for diagnosis of lesions of the cervical spinal cord. DWI was performed using a technique in which echo collection after the application of motion-probing gradients was done in the same manner as in the single-shot fast spin-echo technique. We first imaged six healthy volunteers to demonstrate the cervical spinal cord using the sequence. Then we applied the sequence to 12 patients with cervical myelomalacia due to chronic cord compression. The spinal cord was well seen in all subjects without the distortion associated with echo-planar DWI. In the patients, lesions appeared as areas of low- or isointense signal on DWI. Calculated apparent diffusion coefficients of the lesions (3.30+/-0.38x10(-3) mm(2)/s) were significantly higher than those of normal volunteers (2.26+/-0.08x10(-3) mm(2)/s). Increased diffusion in areas of cervical myelomalacia, suggesting irreversible damage, can be detected using this technique. PMID- 12592492 TI - Sinonasal Wegener's granulomatosis: CT characteristics. AB - Wegener's granulomatosis (WG) is a severe and potentially lethal granulomatosis. Even though no specific radiological criteria exist, CT may suggest the correct diagnosis at an early stage. Recent improvement in the prognosis is related to earlier diagnosis, allowing the initiation of efficient and specific treatment before any severe complications occur. We reviewed a series of WG cases in order to establish the CT diagnostic criteria. PMID- 12592493 TI - Acute encephalitis associated with measles: MRI features. AB - We document the MRI features in six patients aged 5-14 years with acute encephalitis following measles. The diagnosis was made on a characteristic morbiliform rash and detection of specific IgM and IgG antibodies. The symptoms of encephalitis occurred 1-11 days after the appearance of the rash. All patients underwent MRI within 1-4 days of the onset of neurological symptoms. Diffusion weighted images (DWI) were obtained in three patients. In all patients, T2 weighted images showed widely distributed, multifocal high signal in both cerebral hemispheres with swelling of the cortex, with bilateral, symmetrical involvement of the putamen and caudate nucleus. The lesions had showed low apparent diffusion coefficients. Three patients showed subacute gyriform haemorrhage, and asymmetrical gyriform contrast enhancement on follow-up MRI. PMID- 12592494 TI - A case of Salla disease with involvement of the cerebellar white matter. AB - Salla disease (SD) is a lysosomal disorder manifesting in infancy with hypotonia, nystagmus, ataxia and retarded motor development. MRI typically shows hypomyelination confined to the cerebral white matter. We describe a patient with two MRI studies in addition to repeated urine examinations. This case was problematic because the first urine examination did not show the elevation of free sialic acid typical of SD and MRI was also atypical, with abnormal signal intensity in cerebellar white matter. We recommend repeated urinary examinations and a search for SLC17A5 mutations in patients with cerebral signal intensity abnormalities typical of SD and emphasise that cerebellar white-matter involvement on MRI does not exclude the diagnosis. PMID- 12592495 TI - Embolisation of spinal dural arteriovenous fistulae with Onyx. AB - Surgery is currently the standard treatment for spinal dural arteriovenous fistulae (DAVF). Endovascular embolisation of these lesions using N-butyl cyanoacrylate (NBCA) has a high success rate. Onyx is a new liquid embolic agent whose viscosity makes it suitable for treatment of spinal DAVF where penetration into the proximal radicular vein is required. It is delivered with greater control than NBCA without the necessity for rapid withdrawal of the catheter and may therefore overcome some of the drawbacks of NBCA. We report two patients who underwent Onyx embolisation of spinal DAVF; to our knowledge this is the first such report. PMID- 12592498 TI - Modulation of intracortical neuronal circuits in human hand motor area by digit stimulation. AB - We investigated the changes in intracortical neuronal circuits of the hand motor cortex following sensory stimulation of the fingers in 11 healthy subjects. Motor evoked potentials (MEPs) were recorded from intrinsic hand muscles (right first dorsal interosseous and abductor digiti minimi muscles). Electrical stimulation was applied to a digit near (homotopic) or distant (heterotopic stimulation) from each muscle. The right index or little finger was stimulated electrically, followed by single- or paired-pulse transcranial magnetic stimulation (TMS) at an interval of 25, 200, 600, 1,000 or 1,400 ms. Paired-pulse TMS was applied with interstimuli intervals of 2 ms or 12 ms and was expected to stimulate inhibitory or facilitatory intracortical circuits, respectively. MEPs induced by single pulse TMS were significantly suppressed 200, 600, and 1,000 ms after heterotopic and homotopic stimuli. Intracortical facilitation was significantly enhanced only after homotopic stimuli and such enhancement was maximal 200 ms after digit stimulation. Intracortical inhibition was slightly weakened after homotopic stimulation but this effect did not reach statistical significance ( P=0.25). Our results show that sensory feedback can modify intracortical and corticospinal motor excitability and that intracortical facilitation can be enhanced in a topographic-specific way especially at long latencies. These findings suggest that indirect pathways, probably through somatosensory cortex and other areas, enhance intracortical motor excitability in a somatotopically organized manner. PMID- 12592499 TI - Focal reduction of intracortical inhibition in the motor cortex by selective proprioceptive stimulation. AB - The influence of proprioception on motorcortical excitability was assessed by muscle vibration (MV; 80 Hz, 0.5 mm amplitude) of the flexor carpi radialis muscle (FCR) and compared to voluntary contraction and relaxation conditions. Motor thresholds, motor-evoked potentials (MEPs) in response to single pulses of transcranial magnetic stimulation (TMS) and the intracortical inhibition (ICI) and facilitation (ICF) after paired magnetic stimuli were studied. A control experiment using TMS inducing posteriorly directed current was performed. MEPs were recorded simultaneously from the FCR, the extensor carpi radialis, the abductor pollicis brevis and the first dorsal interosseus. In the FCR, MV led to an increase of excitability shown by a decrease of motor threshold, a facilitation of MEPs in response to single-pulse TMS, a reduction of ICI and an increase of ICF. Since especially the ICI and ICF remain unchanged in other recorded muscles, this increase of excitability is specific for the vibrated muscle. With posteriorly directed current the ICI in the FCR was reduced as well, showing an involvement of later I-waves. We suggest that MV induces a focused motorcortical activation which relies on a reduced activity of intracortical inhibitory interneuronal circuits targeting selectively the motorcortical representation of the vibrated muscle. PMID- 12592500 TI - Control of ball-racket interactions in rhythmic propulsion of elastic and non elastic balls. AB - Ball-racket interactions were investigated in a task where participants propelled a ball rhythmically into the air. The study contrasted two ball-racket conditions: (1) an elastic impact where the ball was able to rebound due to the elasticity of the colliding objects and participants bounced the ball, and (2) a non-elastic impact where the coefficient of restitution was zero and the ball did not rebound such that the participants had to throw the ball. The goal of the study was to contrast the situations where haptic information about the ball racket interactions is either secondary (elastic bouncing) or becomes a primary factor for control (non-elastic propulsion). In the elastic condition, the performers controlled the parameters for ball-racket contact prior to contact: In agreement with the criteria for dynamical stability defined by a model, racket accelerations immediately before the contacts were negative, racket positions and velocities at the instant of the initial contact correlated negatively, contact durations were short (30+/-9 ms), and during the collision interval racket velocity and acceleration decreased monotonically. In the non-elastic condition, the parameters of ball release were primarily controlled during the collision phase: Racket accelerations before contact were positive, racket positions and velocities at initial contact showed weak correlations, and the contact intervals were significantly longer (116+/-15 ms) with a clear segmentation into two segments. Negative correlations were observed between the integrals of the velocity and acceleration computed over the two consecutive segments, giving evidence that in the non-elastic condition the CNS is able to introduce corrections during the very short collision interval. The results are discussed with respect to physiological mechanisms of movement corrections available during such short time intervals. PMID- 12592501 TI - Memory-motor transformations are impaired in Parkinson's disease. AB - Parkinson's disease patients are known to suffer loss of dopaminergic input to the rostral caudate nucleus. Recent functional magnetic resonance imaging (fMRI) studies have implicated this structure in the transformation of spatial information in memory to guide action, suggesting that memory to motor transformations may be selectively impaired in Parkinson's disease. In order to investigate this possibility we tested a group of Parkinson's disease patients (PDs) using a memory-guided pointing task. Of interest was whether patients showed reduced accuracy in the task as a function of memory load. Twelve PD patients and 13 elderly controls were asked to recall single or four step target sequences with 2 time delays (500 and 3,500 ms). In all memory-guided conditions PD patients showed increased variability in memory-guided movement end-points. This effect was not affected by delay, number of items, or the sequence familiarity. The results are consistent with increased variability in memory motor transformations in early PD, due to dopamine depletion within the rostral caudate nucleus. PMID- 12592502 TI - Kinesthetic perception of visually specified axes. AB - The purpose of this research was to determine whether human subjects could align the forearm more accurately to the orientation of an external object than to earth-fixed vertical and trunk-fixed anterior-posterior (a-p) axes. Ten young adults aligned the unseen forearm to earth-fixed vertical and trunk-fixed a-p axes, and to a visually presented rod (external visual axis) held by an experimenter in various oblique vertical and horizontal orientations. The head and trunk orientations were varied by left/right lateral flexion when aligning the forearm to vertical plane axes and by rotation about the vertical axis when aligning the forearm to horizontal plane axes. Perceptual errors for aligning the forearm to vertical plane axes were much lower when aligning the forearm to earth fixed vertical than to an external visual axis positioned in a vertical plane. Furthermore, the perceptual errors for aligning the forearm to the visually presented rod were correlated with rod orientation while errors for aligning the forearm to vertical while viewing the rod were unaffected by rod orientations. Clearly, human subjects cannot use an oblique external visually presented axis to provide a frame of reference for accurate perception of forearm orientation in vertical planes. Perceptual errors were similar for aligning the forearm to the horizontal trunk-fixed a-p axis and external visual axis when head and trunk orientation were varied. These perceptual errors were not correlated with rod orientation in the horizontal plane, giving no evidence of bias toward the trunk or external visual axis in horizontal plane perception of forearm orientation. Thus, humans can use either the trunk-fixed a-p axis or the visually specified orientation of an external object as a frame of reference for the kinesthetic system to specify forearm orientation in the horizontal plane. PMID- 12592503 TI - Encoding, learning, and spatial updating of multiple object locations specified by 3-D sound, spatial language, and vision. AB - Participants standing at an origin learned the distance and azimuth of target objects that were specified by 3-D sound, spatial language, or vision. We tested whether the ensuing target representations functioned equivalently across modalities for purposes of spatial updating. In experiment 1, participants localized targets by pointing to each and verbalizing its distance, both directly from the origin and at an indirect waypoint. In experiment 2, participants localized targets by walking to each directly from the origin and via an indirect waypoint. Spatial updating bias was estimated by the spatial-coordinate difference between indirect and direct localization; noise from updating was estimated by the difference in variability of localization. Learning rate and noise favored vision over the two auditory modalities. For all modalities, bias during updating tended to move targets forward, comparably so for three and five targets and for forward and rightward indirect-walking directions. Spatial language produced additional updating bias and noise from updating. Although spatial representations formed from language afford updating, they do not function entirely equivalently to those from intrinsically spatial modalities. PMID- 12592505 TI - Movement-related modulation of vibrotactile detection thresholds in the human orofacial system. AB - By virtue of the direct coupling between circumoral skin and the underlying orofacial musculature, mechanosensation associated with precise orofacial force control may contribute significantly to processes associated with perception, proprioception, and sensorimotor control in this region. The purpose of this study was to assess lower lip (LL) vibratory detection thresholds of adult subjects during the simultaneous performance of a visually guided and continuous lip motor control task. Vibrotactile inputs were delivered to the right LL vermilion at test frequencies of 5, 10, 50, 150, 250, and 300 Hz. The psychophysical detection task was performed simultaneously with the three force control conditions: a null-force baseline condition, an active force control task performed with the right index finger, and an active force control task performed with the lip musculature. For the active tasks, subjects were instructed to use their analog force signal (lip or finger) to continuously perform a visually guided precision force task by tracking a 2 Hz sinusoidally moving target calibrated to a peak-to-peak force load of 0.2 N. Both the analog force signal and the target-tracking signal were displayed in real-time on an oscilloscope. Results showed a statistically significant elevation of LL vibrotactile detection thresholds for test frequencies below 50 Hz during the simultaneous performance of the lip force control task. Disassociating the site of motor control from the location of sensory stimulation (active control task) was effective in normalizing the elevations in LL vibrotactile thresholds, thus demonstrating that the threshold elevation during the lip force task was not solely an artifact of the added attentional load resulting from combining a perceptual task with a motor control requirement. These findings are discussed in relation to published reports of movement-related sensory gating in limb systems and the possible significance that this phenomenon may have for perception and proprioception in the orofacial system. PMID- 12592504 TI - Preparatory states in crossmodal spatial attention: spatial specificity and possible control mechanisms. AB - We used event-related functional magnetic resonance imaging to study the neural correlates of endogenous spatial attention for vision and touch. We examined activity associated with attention-directing cues (central auditory pure tones), symbolically instructing subjects to attend to one hemifield or the other prior to upcoming stimuli, for a visual or tactile task. In different sessions, subjects discriminated either visual or tactile stimuli at the covertly attended side, during bilateral visuotactile stimulation. To distinguish cue-related preparatory activity from any modulation of stimulus processing, unpredictably on some trials only the auditory cue was presented. The use of attend-vision and attend-touch blocks revealed whether preparatory attentional effects were modality-specific or multimodal. Unimodal effects of spatial attention were found in somatosensory cortex for attention to touch, and in occipital areas for attention to vision, both contralateral to the attended side. Multimodal spatial effects (i.e. effects of attended side irrespective of task-relevant modality) were detected in contralateral intraparietal sulcus, traditionally considered a multimodal brain region; and also in the middle occipital gyrus, an area traditionally considered purely visual. Critically, all these activations were observed even on cue-only trials, when no visual or tactile stimuli were subsequently presented. Endogenous shifts of spatial attention result in changes of brain activity prior to the presentation of target stimulation (baseline shifts). Here, we show for the first time the separable multimodal and unimodal components of such preparatory activations. Additionally, irrespective of the attended side and modality, attention-directing auditory cues activated a network of superior frontal and parietal association areas that may play a role in voluntary control of spatial attention for both vision and touch. PMID- 12592506 TI - Early- and late-responding cells to saccadic eye movements in the cortical area V6A of macaque monkey. AB - The cortical area V6A, located in the dorsal part of the anterior bank of the parieto-occipital sulcus, contains retino- and craniocentric visual neurones together with neurones sensitive to gaze direction and/or saccadic eye movements, somatosensory stimulation and arm movements. The aim of this work was to study the dynamic characteristics of V6A saccade-related activity. Extracellular recordings were carried out in six macaque monkeys performing a visually guided saccade task with the head restrained. The task was performed in the dark, in both the dark and light, and sometimes in the light only. The discharge of certain neurones during saccades is due to their responsiveness to visual stimuli. We used a statistical method to distinguish responses due to visual stimulation from those responsible for saccadic control. Out of 597 V6A neurones tested, 66 (11%) showed responses correlated with saccades; 26 of 66 responded also to visual stimulation and 31 of 66 did not; the remaining 9 were not visually tested. We calculated the response latency to saccade onset and its inter-trial variance in 24 of 66 neurones. Saccade neurones could respond before, during or after the saccade. Neurones responding before saccade-onset or during saccades had much higher latency variance than neurones responding after saccades. The early-responding cells had a mean latency (+/-SD) of -64+/-62 ms, while the late-responding cells a mean latency of +89+/-20 ms. The responses to saccadic eye movements were directionally sensitive and varied with the amplitude of the saccade. Responses of late-responding cells disappeared in complete darkness. We suggest that the activity of early-responding cells represents the intended saccadic eye movement or the shift of attention towards another part of the visual space, whereas that of late-responding cells is a visual response due to retinal stimulation during saccades. PMID- 12592507 TI - Electromyographic and contractile properties of rabbit masseter motor units during fatiguing stimulation. AB - Trigeminal motoneurons were electrically stimulated in order to investigate the electromyographic (EMG) behavior in relation to the contractile properties of motor units of the masseter muscle. A total of 80 motor units were studied in situ in male New Zealand White rabbits ( n=46). The motor units were separated into two groups, each exposed to a specific fatiguing stimulation regimen. Motor unit action potential (MUAP) features, which comprised the amplitude (AMP) and inter-peak time (IPT), and the tetanic force were measured. All motor units were classified as fast (F) units. Forty-one motor units underwent a prolonged standard fatigue regimen of 40-Hz trains at 1 Hz for 20 min. While the MUAP showed an immediate decrease of mean AMP at the beginning of the stimulation, the mean force and IPT increased. After 2 min, the force declined, while the IPT continued to increase until 20 min. Only after 3 min of stimulation, did the degree of force decrease parallel the decline in MUAP AMP. After 20 min of stimulation, the majority of motor units ( n=34) still generated a force larger than 50% of the initial value, but only 17 motor units showed MUAP AMP of less than 50% of the initial EMG response. A more intensive fatigue regimen (40-Hz trains at 1.5 Hz) was applied to another group of 39 motor units. A rapid decline of force and MUAP amplitude to almost 50% was observed within the first 5 min of stimulation. After 20 min, only four motor units were still able to produce a tetanic force of more than 50% of the initial. Most strikingly, motor units with twitch contraction times faster than 22 ms exhibited a decrease in force more than in MUAP AMP, whereas the reverse was seen for units slower than 22 ms; motor units with a twitch contraction time of 22 ms showed equal decrease in AMP and force. This finding is suggestive of a division of fast masseter motor units into two classes, those which fatigue more rapidly mechanically and those which fatigue more readily electrically. PMID- 12592509 TI - Characterisation of axon terminals in the rat dorsal horn that are immunoreactive for serotonin 5-HT3A receptor subunits. AB - Serotonin 5-HT(3) receptors are abundant in the superficial dorsal horn and are likely to have an involvement in processing of nociceptive information. It has been shown previously that 5-HT(3) receptors are present on primary afferent terminals and some dorsal horn cells. The primary aim of the present study was to determine what classes of primary afferent possess 5-HT(3)A receptor subunits. We performed a series of double- and triple-labelling immunofluorescence experiments. Subunits were labelled with an anti-peptide antibody and primary afferent axons were identified by the presence of calcitonin gene-related peptide (CGRP) and binding of the lectin IB4. Quantitative confocal microscopic analysis revealed that approximately 10% of axons displaying 5-HT(3)A immunoreactivity were also labelled for CGRP but that only 3% of these fibres bind IB4. We also investigated the relationship between immunoreactivity for the subunit and descending serotoninergic systems, axons originating from inhibitory neurons that contain glutamic acid decarboxylase, and axons of a subpopulation of excitatory neurons that contain neurotensin. None of these types of axon was associated with immunoreactivity for receptor subunits. Ultrastructural studies confirmed that punctate immunoreactive structures observed with the light microscope were axon terminals. These terminals invariably formed asymmetric synaptic junctions with dendritic profiles and often contained a mixture of granular and agranular vesicles. Some terminals formed glomerular-like arrangements. Immunoreactive cells were also examined and were found to contain intense patches of reaction product within the cytoplasm. We conclude that the majority (about 87%) of dorsal horn axons that are immunoreactive for 5-HT(3)A receptor subunits do not originate from the subtypes of primary afferent fibres that bind IB4 or contain CGRP. It is likely that most of these axons have an excitatory action and they may originate from dorsal horn interneurons and/or fine myelinated primary afferent fibres. PMID- 12592508 TI - Persistent effects of high frequency repetitive TMS on the coupling between motor areas in the human. AB - Repetitive transcranial magnetic stimulation (rTMS) shows promise as a treatment for various movement and psychiatric disorders. How rTMS may have persistent effects on cortical function remains unclear. We hypothesised that it may act by modulating cortico-cortical connectivity. To this end we assessed cortico cortical coherence before and after high frequency rTMS of the motor cortex. Sixteen healthy subjects received a single train (5 Hz, active motor threshold, 50 stimuli) of rTMS to the left motor hand area. Spectral power and coherence estimates were calculated between different EEG signals at rest and while muscles of the distal upper limb were tonically contracted. Repetitive TMS over the left motor hand area caused a significant decrease in the intrahemispheric EEG-EEG coherence between motor and premotor cortex in the 10.7-13.6 Hz (upper alpha band) lasting a few minutes after stimulation. There was no significant change in interhemispheric EEG-EEG coherence between motor areas. Thus, high frequency rTMS of the motor cortex decreases ipsilateral cortico-cortical intrahemispheric in the upper alpha band. PMID- 12592510 TI - Inhibition of return in children with attention deficit hyperactivity disorder. AB - Earlier studies have suggested an impairment in the attention and eye movement control of children with ADHD. An important phenomenon in the control of attentional shifts and eye movements is the inhibition of return (IOR), which states that our brain works in a way that prevents our attention from returning to a spatial location that has been attended to, either overtly or covertly. This current study addresses whether the IOR in oculomotor planning is compromised in children with ADHD. Eleven ADHD and 12 age- and gender-matched control subjects participated in a behavioral task, in which they made saccades to a peripheral target after a valid, invalid or neutral cue. The latency difference between cued and uncued saccades over a range of cue-target onset asynchrony as well as the positive component of this latency profile (i.e., IOR) was compared between groups. The results show that ADHD children demonstrate a biphasic latency profile that is grossly similar to that observed in control subjects, although the magnitude of IOR appears to be slightly smaller in ADHD subjects. These preliminary results suggest that the inhibitory attention mechanism subserving IOR is at least not fully compromised in ADHD children. PMID- 12592511 TI - Dentine-lead levels and dental caries in First Nation children from the western James Bay region of northern Ontario, Canada. PMID- 12592512 TI - Use of manganese as tracer in the determination of respiratory exposure and relative importance of exposure routes in the safety of pesticide applicators in citrus orchards. PMID- 12592513 TI - Organic pollution of street dust in the Handan City, China. PMID- 12592514 TI - Plasma nitrogen oxides levels in taxi drivers and community residents. PMID- 12592515 TI - Contamination of aluminium from cooking utensils and yogurt containers. PMID- 12592516 TI - Toxic bromate residues in Nigerian bread. PMID- 12592517 TI - General chemical water quality of private groundwater supplies in Saskatchewan, Canada. PMID- 12592518 TI - Bioremediation of organophosphate pollutants by a genetically-engineered enzyme. PMID- 12592519 TI - Phytoremediation of lead, nickel, and copper by Salix acmophylla Boiss.: role of antioxidant enzymes and antioxidant substances. PMID- 12592520 TI - Comparative deposition of diphenyl diselenide in liver, kidney, and brain of mice. PMID- 12592521 TI - Impact of herbicide application rates and crop residue type on earthworm weights. PMID- 12592522 TI - Organochlorine pesticide residues in water and fish samples: first report from rivers and streams of Kumaon Himalayan region, India. PMID- 12592523 TI - Sediment toxicity in Central and South Florida ecosystems. PMID- 12592524 TI - Heavy metals and radionuclides in lichens and mosses in Thrace, Turkey. PMID- 12592525 TI - Metal concentrations in tissues of common hamsters (Cricetus cricetus [L.]) from an agricultural area in Germany. PMID- 12592526 TI - Estimation of the sorption of substituted aromatic compounds onto modified clay. PMID- 12592527 TI - Degradation of ethoprophos in three physicochemically different soils treated with composted manure and chemical fertilizer. PMID- 12592528 TI - Nonylphenol ethoxylates and their biodegradation intermediates in water and sludge of a sewage treatment plant. PMID- 12592529 TI - Toxicity of the insecticides fipronil and endosulfan to selected life stages of the grass shrimp (Palaemonetes pugio). PMID- 12592530 TI - Comparative acute toxicity of herbicides to photosynthesis of coral zooxanthellae. PMID- 12592531 TI - Structure-toxicity relationships for methyl esters of cyanoacetic acids to Tetrahymena pyriformis. PMID- 12592532 TI - Correlation between heavy metal toxicity and metal fractions of contaminated soils in Korea. PMID- 12592533 TI - Cadmium accumulation and its phytotoxicity in Potamogeton pectinatus L. (Potamogetonaceae). PMID- 12592534 TI - Response of fast growing woody plants from family Salicaceae to cadmium treatment. PMID- 12592535 TI - Persistence and effectiveness of iprodione against Alternaria blight in mustard. PMID- 12592536 TI - Toxicity of cadmium-copper-nickel-zinc mixtures to larval purple sea urchins (Strongylocentrotus purpuratus). PMID- 12592537 TI - Effects of ethylene and propylene glycol on development and hatching success in the medaka, Oryzias latipes. PMID- 12592538 TI - Monitoring chlorophyll- a as a measure of algae in Lake Texoma Marinas. PMID- 12592539 TI - Induction of mortality and malformation in Scinax nasicus tadpoles exposed to glyphosate formulations. PMID- 12592540 TI - Natural occurrence of metallothionein-like proteins in the liver of fish Oreochromis niloticus and effects of cadmium, lead, copper, zinc, and iron exposures on their profiles. PMID- 12592541 TI - Pain-drawing does not predict the outcome of fusion surgery for chronic low-back pain: a report from the Swedish Lumbar Spine Study. AB - Despite a widespread use of pain drawing in the selection of patients for surgical and non-surgical treatment, its value as a predictor of outcome is still not well documented. In a prospective multicentre randomised controlled trial of surgical and non-surgical treatment for chronic low-back pain (CLBP), two hypotheses were tested: (1). Pain drawing predicts outcome of treatment for CLBP, (2). Pain drawing is associated with psychological characteristics of patients with CLBP. Two hundred and sixty-four patients with severe CLBP of long duration completed pain drawings as part of a battery of questionnaires prior to treatment. They were followed up at 2 years post-treatment, with renewed completion of questionnaires. Outcome was measured in three ways: patient global assessment, change of disability/pain, and work status. The pain drawing was analysed by four different methods. The association between the pain drawings and outcomes was analysed. Personality traits and depressive symptoms were evaluated in the psychological assessment. None of the four methods of interpretation of the pain drawings demonstrated any significant association with outcome, in either the surgical or the non-surgical group. The pain drawing was associated with pre-treatment back pain intensity and depressive symptoms. No predictive value of the pain drawing regarding the outcome of treatment of CLPB was demonstrated. The concept of "organic/non-organic" pain in conjunction with chronic low-back pain is not supported by the results of the present study. PMID- 12592542 TI - The clinical importance of changes in outcome scores after treatment for chronic low back pain. AB - When measuring treatment effect in chronic low back pain with multi-item outcome instruments, it is necessary, both for clinical decision-making and research purposes, to understand the clinical importance of the outcome scores. The aims of the present study were three-fold. Firstly, it aimed to estimate the minimal clinically important difference of three multi-item outcome instruments (the Oswestry Disability Index, the General Function Score and the Zung Depression Scale) and of the visual analogue scale (VAS) of back pain. Secondly, it aimed to estimate the error of measurement of these instruments; and its third aim was to describe the clinical meaning of score change. The study population consisted of 289 patients treated surgically or non-surgically in a randomised controlled trial. The minimal clinically important difference was estimated with patient global assessment as the external criterion. It was compared with the standard error of measurement of the instruments. The individual items of the instruments were compared for score changes related to improvement and deterioration. The standard error of measurement of the Oswestry Disability Index, the General Function Score and the Zung Depression Scale was 4, 6 and 3 units, respectively. The 95% tolerance interval was 10, 16 and 8 units, respectively. The minimal clinically important difference was 10, 12 and 8-9 units, respectively, thus not significantly exceeding the tolerance interval. The minimal clinically important difference of VAS back pain was 18-19 units, well exceeding the 95% tolerance interval, which was 15 units. Improvement after treatment for chronic low back pain tends to occur to a greater extent in sleep disturbance, ability to do usual things and psychological irritability, but to a lesser extent in the ability to sit, stand and lift. We conclude that the VAS of back pain is responsive enough to detect the minimal clinically important difference, whereas the smallest acceptable score changes of the Oswestry Disability Index, the General Function Score and the Zung Depression Scale may require an increase to exceed the 95% tolerance interval when used for clinical decision making and for power calculation. Despite improvement after treatment, the ability to sit, stand and lift, remain notable problems. PMID- 12592544 TI - Predictors of outcome in fusion surgery for chronic low back pain. A report from the Swedish Lumbar Spine Study. AB - Despite the continuous development of surgical techniques and implants, a substantial number of patients still undergo surgery for chronic low back pain (CLBP) without any benefit, or even become worse. With the aim of finding predictors of functional and work status outcome, 264 patients with severe CLBP of long duration, randomised to surgical or non-surgical treatment, were characterized by socio-demographic, clinical, radiological and psychological variables. The variables were estimated as predictors of outcome at the 2-year follow-up. Univariate and multiple logistic regression analyses were used in both treatment groups. We found that a personality characterized by low neuroticism and low disc height were significant predictors of functional improvement after surgical treatment. Depressive symptoms predicted functional improvement after non-surgical treatment. Work resumption was predicted by low age and short sick leave in the surgical group, and by short sick leave in the non-surgical group. We conclude that improved selection of successful surgical candidates with CLBP seems to be promoted by attention to severe disc degeneration, evaluation of personality traits and shortening of preoperative sick leave. PMID- 12592545 TI - The use of poly-L-lactic acid in lumbar interbody cages: design and biomechanical evaluation in vitro. AB - Cage design and cage material may play a crucial role in the incidence of postoperative complications reported with current non-absorbable interbody cage devices. Bioabsorbable poly-L-lactic acid cage devices may have potential benefits. The purpose of this study was to determine the required strength of poly-L-lactic acid cages for use in experimental goat studies and to evaluate the mechanical properties of different cage designs in situ. The yield and ultimate strength of native goat motion segments (L1-L6) were determined; the yield strength was used as a design parameter for the cages. The mechanical behaviour of two types of poly-L-lactic acid cages, the influence of endplate perforation, differences between toothed and smooth cages, and the influence of cage filling were biomechanically tested and compared to native motion segments. Only axial compression until failure of the motion segments was performed. Dual energy X-ray absorptiometry was used to determine bone mineral content. The yield and ultimate strength of the native motion segments were 3.5 and 7.0 kN, respectively. Based on these data, flexible and stiff poly-L-lactic acid cages were designed with strengths of 3.5 and 7 kN, respectively. Poly-L-lactic acid cages, whether with or without bone graft and perforating the endplates, did not reduce the compressive strength of motion segments as compared to native segments. However, toothed titanium cages, with the same geometry, negatively influenced the segments' compressive strength, which effect was reduced using smooth titanium cages. PMID- 12592546 TI - Temporary external pedicular fixation versus definitive bony fusion: a prospective comparative study on pain relief and function. AB - Temporary external pedicular fixation is used as a prognostic instrument when treating degenerative conditions with spinal fusion. We studied the validity of the method and whether a functional test could improve the prognostic value of such fixation. Twenty-six patients with long-standing lumbar pain had an external temporary fixation. Pain levels were registered before fixation on a visual analogue scale at rest, as a mean for the previous week, and at seven different standardized activities. Walking capacity and walking speed for a standardized distance were also measured. Identical evaluations were then repeated during the external fixation and 1 year after definitive fusion. Based on the outcome of the temporary fixation, 20 patients were recommended for definitive surgical fusion. In six cases, the option of fusion surgery was rejected due to an unfavourable pain response or insufficient pain relief during the test fixation period, and this group was not further followed within the study. One year after surgery, 14 of 20 patients reported a good outcome. Solid bony fusion assessed by conventional radiography was seen in 19 patients. One patient with a poor clinical outcome had a pseudarthrosis. The mean values for pain level at rest, during last week and at the seven different activities in the functional test tended to decrease after fusion compared to the situation with temporary external fixation. In no activity did the external fixator overestimate the mean positive pain-relieving effect after definitive fusion. The walking capacity significantly increased, while the walking speed did not alter at the three different measurements. We conclude that with a good outcome ratio of 14 patients out of 19 having a solid fusion, the external frame improved patient selection and can be used as a valid prognostic instrument. The pain relief and function after definitive fusion can not be quantified by the external fixation, probably due to the fact that the stabilisation with an external frame is partial. The value of the functional test design presented is moderate, and an outcome evaluation comprising pain relief at rest and mean pain level during a week in fixation seems adequate. PMID- 12592547 TI - Arterial complications following anterior lumbar interbody fusion: report of eight cases. AB - The objective of this study is to report eight cases of arterial complication following anterior lumbar interbody fusion (ALIF) and to analyze the data in order to identify possible risk factors. The authors have encountered six cases of common iliac artery occlusion and two cases of acute vasospasm as a complication of ALIF using two different approaches to spine: hypogastric-midline transperitoneal approach at one center and minimally invasive muscle-sparing retroperitoneal approach at the other. All cases involved surgery at the L4-L5 level. All patients were smokers, and three had an existing history of vascular disease. The left iliac and common femoral arteries were involved in seven cases, while the right common iliac was involved in one case. Five patients had thromboembolism, one patient had an intimal tear and two had functional vasospasm. Circulation to the lower limb was restored by thrombectomy (five patients) and arterioplasty for the intimal tear (one patient). One of the vasospasm cases was explored (false-positive), while the other was treated conservatively. One of the patients with thrombosis developed rhabdomyolysis resulting in fatal acidosis. All but the first case at each center was diagnosed either intraoperatively or within 2 h of surgery. We believe that awareness of this potentially serious complication will lead to precautionary measures for prevention of the problem as well as early diagnosis and management of the complication if it does occur. Intraoperative monitoring of lower limb blood flow by measuring the toe oxygenation with a pulse oxymeter can prove to be helpful in early diagnosis. PMID- 12592548 TI - Assessment of the cellular heterogeneity of the ovine intervertebral disc: comparison with synovial fibroblasts and articular chondrocytes. AB - The objective of this study was to compare and contrast the cellular heterogeneity of ovine intervertebral disc (IVD) annulus fibrosus (AF) and nucleus pulposus (NP) cells, synovial fibroblasts (SFs) and articular chondrocytes (ACs) grown in monolayer and alginate bead culture. Phase contrast microscopy of the monolayers indicated that the SF and AF cells, and the AC and NP cells had similar morphologies. Immunolocalisation of type I, II and VI collagen epitopes in the monolayers, however, demonstrated distinct quantitative and relative differences between the SF and AF, and the AC and NP cells. Immunolocalisation of bead-incorporated proteoglycans (PGs) also demonstrated quantitative and qualitative differences between the SF and AF cells. Quantitation of (35)S-bead PGs indicated that the AF cells synthesised significantly higher PG levels than the SFs, and AC cells synthesised significantly higher PG levels than the NP cells. These data were also consistent with the relative metabolic activities and cellular viabilities displayed by each cell type in bead culture. AF cells shared morphological traits with SFs in monolayer, but displayed distinctly different characteristics in bead culture. Similarly, NP cells shared similarities with ACs in monolayer and bead culture, but overall were less metabolically active. Data presented here support the proposal that the AF and NP cells of the ovine IVD should be considered as specialised fibrochondrocytic cells. PMID- 12592549 TI - Induction of matrix metalloproteinase-2 and -3 activity in ovine nucleus pulposus cells grown in three-dimensional agarose gel culture by interleukin-1beta: a potential pathway of disc degeneration. AB - Degeneration of the intervertebral disc is an important clinical problem, which often contributes to low back pain. Since approximately 80-90% of the general population will be subject to back pain at some stage during their lifetime, this has major socioeconomic consequences. Matrix metalloproteinases (MMPs) have been implicated in the excessive breakdown of extracellular matrix components during disc degeneration. The aim of the present study was to evaluate the regulation of MMP-2 (gelatinase-A) and MMP-3 (stromelysin) produced by cultured ovine nucleus pulposus (NP) cells stimulated with interleukin-1beta (IL-1beta). NP cells were established in three-dimensional agarose culture and stimulated with IL-1beta under serum-free conditions. Conditioned media samples were evaluated by gelatin and casein zymography and by fluorimetry using an MMP-specific substrate. Time course and dose dependencies were established for MMP-2, -3 production by the NP cells in response to the IL-1beta. Gelatin and casein zymography indicated that elevated levels of proMMP-2 and proMMP-3 were present in media samples in response to the IL-1beta treatment. After 24-96 h culture, levels of the active 43 and 45 kDa active MMP-3 were significantly elevated, whereas MMP-2 was present mainly as its 72 kDa pro-form. Additional 36, 28 and 21 kDa MMP species were also present after prolonged incubation with IL-1beta, probably representing MMP breakdown species. IL-1beta was a potent catabolic mediator for the NP cells, resulting in the production of elevated levels of MMP-2 and -3 in culture. However, approximately 70% of the MMP-2 was present as the 72 kDa pro-form, which suggests that some additional steps are involved in its activation in vivo. PMID- 12592550 TI - Intradiscal application of hyaluronic acid in the non-human primate lumbar spine: radiological results. AB - Prospectively, with randomized segment-treatment assignment, and with blinded evaluators, lumbar motion segments in Cercopithecus monkeys were analyzed for macroscopic and radiological changes 24 weeks after nucleotomy and nucleotomy with additional intradiscal application of different hyaluronic acid formulations versus untreated control segments. The objective was to find out whether hyaluronic acid is able to influence the degenerative cascade in nonhuman primates after nucleotomy. In a similar procedure, hyaluronic acid has proven to decrease degeneration after nucleotomy in a Minipig model. This is the first such study ever undertaken in primates, thus trying to overcome the known limitations of non-primate spine models. Twenty monkeys with four segments each obtained nucleotomy in three segments and solely exposure of another control segment. Nucleotomy was performed from a transpsoatic retroperitoneal approach. Preoperative radiographs and follow-up radiographs, magnetic resonance imaging (MRI), computed tomography (CT), Q-CT with bone mineral density measurements and three-dimensional reconstruction were obtained and analyzed qualitatively and quantitatively. Segments with high-molecular-weight hyaluronic acid (Hylan G-F 20) application proved to be significantly superior over those with a standard nucleotomy in radiographs, MR images, CT scans, and macroscopic appearance at follow-up. Control segments remained unaffected. Interdependence between the different methods validated the utilized methods of quantitative radiological assessment of degeneration. Hylan G-F 20 appears to be a possible adjunct in reducing postoperative degeneration in an animal nucleotomy model. It deserves further evaluation, despite the fact that the mechanisms of its effects are still speculative. PMID- 12592551 TI - Pharyngolaryngeal lesions in patients undergoing cervical spine surgery through the anterior approach: contribution of methylprednisolone. AB - Cervical spinal surgery through anterior approaches can lead to ear, nose and throat (ENT) complications, some of which, such as obstruction of upper airways by edema, are potentially life threatening. The purpose of this study was to evaluate the incidence of various categories of ENT lesions and to determine whether preventive treatment with methylprednisolone (Mp) is useful in this indication. In a prospective trial, 236 patients who were undergoing anterior cervical spine surgery were separated into two groups, a control group (group I: 158 patients) and a group receiving 1 mg/kg of Mp intravenously at the end of the operation and 12 and 24 h later (group II: 78 patients). All 236 patients underwent nasofibroscopic examination by an ENT specialist the day before the operation and 24 and 36 h after the operation. The lesions observed during these examinations and reported symptoms such as uncomfortable sensation, odynophagia, or dysphagia were compared between the two groups (chi(2) or Mann-Whitney test) in terms of relative risk, with a confidence interval of 95%. Quantitative findings were compared using Student's t-test ( P<0.05). Demographic characteristics and duration of endotracheal intubation were comparable in the two groups. The ENT examination was considered abnormal in 30 patients of group I and 15 patients of group II preoperatively (NS). It was abnormal in 146 patients of group I (92%) and 46 patients of group II (59%) postoperatively. Altered vocal cord motility was noted in six patients of group I and two patients of group II. The relative risk was 1.6 (CI: 1.3-1.9). Postoperatively, 130 patients in group I and 56 of group II (NS) reported unwanted symptoms. The observed lesions were significantly more severe in group I ( P<0.001), involving primarily the pharyngeal wall, the arytenoids, and the vocal cords. The results of this study indicate that ENT complications of anterior cervical spinal surgery are diminished by administration of Mp. Systematic ENT examination is warranted before this type of operation. PMID- 12592552 TI - Posterior atlanto-axial fusion with the Olerud Cervical Fixation System for odontoid fractures and C1-C2 instability in rheumatoid arthritis. AB - In posterior C1-C2 fusion, traditional wire fixation gives poor stability. The bone quality is often insufficient to provide the competent structural bone graft that is required, and the introduction of sublaminar wires is somewhat dangerous. The stability is markedly improved by adding transarticular screws, but the drawbacks of structural bone graft and sublaminar wires remain. The C1 claw of the Olerud Cervical Fixation System improves C1-C2 fixation without relying on structural bone graft or compromising the spinal canal. The aim of this study was to evaluate radiological healing and possible complications in a consecutive series of C1-C2 fusions from our department operated with the C1 claw device. Twenty-six patients (14 women) with a mean age of 73 (range 37-93) years were included. The diagnoses were odontoid fracture in 18 patients, rheumatoid instability in 6, and odontoid non-union and os odontoideum in 1 each. The patients were followed clinically and with plain radiographs for an average of 15 (range 3-27) months. There were no neurological or vascular complications, and no secondary displacements or reoperations in the series. Twenty patients followed for 6-27 months were radiographically healed. Six patients died from unrelated causes 1-38 months postoperatively. Three of these patients had no radiographs later than the postoperative control, one had a healed odontoid fracture but resorbed bone graft at 8 months, while the remaining two patients were not healed, but showed no signs of healing disturbance at the time of death. On the basis of the findings of this study, posterior C1-C2 fusion with the Olerud Cervical Fixation System seems promising. No serious complications related to the surgical procedure were encountered. The stability of the implant obviates the use of a solid bone block as a graft and still allows a high frequency of fusion healing. PMID- 12592553 TI - Severe pathologic compression of three consecutive vertebrae in Gaucher's disease: a case report and review of the literature. AB - To date, only one patient with spinal affection of Gaucher's disease requiring surgery for spinal compression has been reported. We present an additional case with extensive affection of three consecutive vertebral bodies, and present a review of the literature relating to the management of therapy and follow-up of this disease. PMID- 12592554 TI - Chronic spontaneous spinal epidural hematoma -- a rare cause of cervical myelopathy. AB - Spontaneous spinal epidural hematoma (SSEH) is a rare entity. SSEH occurring anterior to the spinal cord in the cervical region with spontaneous improvement is even more rare. One such rare case is reported. A 19-year-old male presented with a 2-month history of neck pain and weakness of the upper and lower limbs without sensory deficit. Magnetic resonance imaging (MRI) showed features of anteriorly located cervical epidural hematoma. As the patient had started improving by the time he sought neurosurgical consultation, he was treated conservatively, and he improved over a period of 2 months. This case illustrates that, though rare, chronic spontaneous spinal epidural hematoma should be entertained in the differential diagnosis of cervical cord compression. Spontaneous resolution of spinal epidural hematomas is known to occur, and in properly selected cases, surgical intervention may not always be necessary. PMID- 12592555 TI - Novel Artemis gene mutations of radiosensitive severe combined immunodeficiency in Japanese families. AB - A subgroup of patients with severe combined immunodeficiency (SCID) and increased cellular radiation sensitivity (RS-SCID) have mutations of Artemis, a gene that encodes a protein essential for V(D)J recombination and DNA double-strand break repair. RS-SCID described to date are either of European origin or are Athabascan speaking native Americans belonging to the Navajo and Apache tribes. We have identified three Japanese boys and one girl from four unrelated families with RS SCID caused by a genomic exon 3 deletion of the Artemis gene, resulting in loss of exon 3 and skipping of exon 4. Two patients were homozygous and two patients were heterozygous for this novel mutation. Those parents studied were heterozygous for this mutation. These findings suggest the genomic exon 3 deletion is unique to Japan and may be considered as a founder haplotype. Although two infants underwent successful bone marrow transplantation and immune reconstitution, the long-term outcome of this procedure is uncertain, because Artemis is expressed in most tissues and lack of its function in cells other than those derived from hematopoietic stem cells may increase the risk of malignancies. PMID- 12592556 TI - More research needed to consolidate achievements in filariasis control. PMID- 12592557 TI - Polymerase chain reaction-based detection of lymphatic filariasis. AB - PCR-based diagnostic assays are promising tools for the monitoring and evaluation of the Global Programme for Elimination of Lymphatic Filariasis. Sensitive and specific assays have been described for the detection of Wuchereria bancrofti, Brugia malayi, or Brugia timori infection in blood, sputum, and vectors. These techniques can be most cost-effective when employed for pool screening, which is important in the later stages of control programs when infection rates of humans and vectors are low, and large numbers of samples must be examined. PMID- 12592558 TI - Lymphatic filariasis in Papua New Guinea: prospects for elimination. AB - Lymphatic filariasis is a significant public health problem in several Pacific island countries. Papua New Guinea is one of the most populous countries in this region, and 39% of its residents are estimated to be infected with Wuchereria bancrofti. The Ministries of Health of the 22 islands and territories in the Pacific region are committed to taking action against lymphatic filariasis. Accordingly, a regional collaborative effort aimed at the control of filariasis has been organized under the auspices of a program referred to as PacELF. The main objective of PacELF is to eliminate filariasis as public health problem in the Pacific region by the year 2010, 10 years before global elimination of this infectious disease has been targeted. This contribution describes the epidemiology and ecological features of filariasis and prospects for its elimination in Papua New Guinea. The frequencies of microfilaremia, chronic lymphatic disease, and acute filarial morbidity in Papua New Guinea are higher than in many other endemic countries of the Pacific, Africa, and South America. All possible combinations of these three manifestations of filariasis exist. They occur independently of each other, and there is no association between chronic lymphatic disease and microfilarial status. Anopheles punctulatus mosquitoes are the main vectors throughout the country. Transmission intensity is heterogeneous and a major determinant of local patent infection and morbidity rates. Annual transmission potential and annual infective biting rates are positively associated with the village-specific microfilarial rate, mean intensity of microfilaremia, and prevalence of leg edema. Children and adults have similar worm burdens, assessed by circulating filarial antigen levels, in areas of high transmission, whereas worm burdens increase with age in areas of lower transmission. Intensity of exposure to infective third-stage larvae (L3) is significantly correlated with filarial antigen-specific lymphocyte proliferation and cytokine production, possibly by a mechanism that alters APC function. Historical evidence suggests that residual insecticide spraying conducted for malaria control in some parts of the country interrupted transmission of W. bancrofti as it did in the Solomon Islands. Prospects for eliminating lymphatic filariasis in Papua New Guinea are good and may be achieved by the end of the second decade of the twenty-first century if an integrated control approach using mass drug administration with vector control is adopted. PMID- 12592559 TI - Epidemiology of concomitant infections due to Loa loa, Mansonella perstans, and Onchocerca volvulus in rain forest villages of Cameroon. AB - An epidemiological survey was conducted in 16 remote villages of the rain forest of southern Cameroon to ascertain the prevalence and intensity of three species of filariae: Loa loa, Onchocerca volvulus, and Mansonella perstans. We examined 1458 individuals for blood-dwelling microfilariae and 1255 of these were also for the presence of palpable nodules. All the villages surveyed were found highly endemic for onchocerciasis and mansonellosis with prevalence ranging from 28.44% to 87.17% for O. volvulus and 52.48% to 100% for M. perstans. The intensities of infection were also found high for M. perstans with arithmetic means of microfilaremia ranging from 280.94 to 4947.57 mf/ml. The loiasis prevalence was relatively low with value from 2.22% to 19.23%. Males were found more infected than females for the three species of filariae, and the prevalence and intensities of microfilaremia vary differently in males and females at different ages. The three species of filariae displayed different degrees of association in the inhabitants with a low prevalence of co-occurrence between L. loa/O. volvulus and between L. loa/M. perstans. In contrast, there was a high prevalence of co occurrence between M. perstans and O. volvulus. The implications of the co occurrence of the three species of filariae in the populations of these remote villages on the intervention programs based on mass treatment with mectizan are discussed. PMID- 12592560 TI - Murine filariasis: interleukin 4 and interleukin 5 lead to containment of different worm developmental stages. AB - We compared the impact of IL-4 and IL-5 deficiency during the fully permissive infection of BALB/c mice with the rodent filaria Litomosoides sigmodontis. IL-5, in contrast to IL-4, is crucial for the containment of adult worms during short- and long-term infections. IL-5 KO mice allowed development of more larvae into adult worms and showed up to 200 times more adult worms persisting during chronic infection (day 60 until 200 post-infection). This increased persistence was accompanied by a reduction in inflammatory nodules around adult filariae. In contrast, adult worm survival and nodule formation did not differ between BALB/c wild-type mice and BALB/c IL-4 KO or BALB/c IL-4 receptor (IL-4R) alpha-chain KO mice. In both IL-4 and IL-5 KO mice microfilaraemia was greatly enhanced (160 fold) and prolonged compared to wild-type mice. This extent of susceptibility to microfilariae required the presence of adult worms in a full infection cycle since upon intraperitoneal injection of microfilariae alone they were removed from BALB/c, BALB/c IL-4 KO and BALB/c IL-4R alpha-chain KO mice with equivalent kinetics, and since microfilarial survival was only slightly increased in IL-5 KO mice (factor of 5 vs. factor of 160 in full infection). In conclusion, IL-4 and IL-5 dependent effector pathways operate against different stages of filarial worms, and IL-5 has a greater impact on parasite containment than IL-4. PMID- 12592561 TI - Cytokine-dependent inflammatory cell recruitment patterns in the peritoneal cavity of mice exposed to the parasitic nematode Brugia malayi. AB - Mice exposed intraperitoneally to either adult or first larval stage (microfilaria) of the human nematode parasite Brugia malayi display polarized cytokine responses. We have used this model to investigate the impact of altered cytokine profiles on inflammatory cell recruitment patterns in vivo. Here we demonstrate that Th2-inducing adult parasites drive the recruitment of eosinophils and macrophages after implant into the murine peritoneal cavity whereas Th1-inducing microfilaria do not. The underlying mechanism of recruitment was further defined by use of mice deficient in the key Th2 cytokines IL-4 or IL5 and mice that lack T cells (nude mice). Recruitment dynamics differed in IL-4 and IL-5 deficient mice, showing reduced or absent eosinophilia. These data emphasize the pivotal role of these cytokines in shaping the cellular profile of inflammatory responses. Surprisingly, the absence of T cells failed to influence inflammatory cell recruitment indicating that recruitment signals are provided by other cell types. PMID- 12592562 TI - Protective immunity in human lymphatic filariasis: problems and prospects. AB - Human filariasis caused by lymphatic dwelling nematodes, affecting 120 million persons worldwide, is a major public health problem. Efforts towards development of vaccines for such large tissue-dwelling nematodes depends significantly on identification and demonstration of protective immunity in the exposed population. Immunological studies conducted in human filariasis so far are essentially attempts to establish a correlation of the immune response phenotypes with presence or absence of filarial infections/disease in the host, and the cause-effect relationship between the observed immune responses in the host and protective immunity continues to be conjectural. This short review attempts to clarify the functional definition of protective immunity, problems associated with identification of putatively immune subjects in endemic areas, role of antibodies reactive to surface of microfilariae and larvae stages of filarial parasites and importance of undertaking immunological investigations on a longitudinal basis in different cohorts of subjects presenting with one or the features of infection and/or disease for more accurate delineation of protective immunity in human filariasis. PMID- 12592563 TI - The FAR proteins of parasitic nematodes: their possible involvement in the pathogenesis of infection and the use of Caenorhabditis elegans as a model system to evaluate their function. AB - Parasitic nematodes secrete a structurally novel class of fatty acid and retinol binding (FAR) protein into the surrounding tissues of the host. These proteins are of interest because they may play an important role in scavenging fatty acids and retinoids from the host that are essential for the survival of the parasite and also because the localised depletion of such lipids may have immunomodulatory effects that compromise the host immune response. Research into the biological function(s) of the FAR proteins has been severely hampered by the difficulties associated with the life-cycle propagation of parasitic nematodes and the current intractability of the parasites to reverse-genetic studies. The genome of the free-living nematode Caenorhabditis elegans, however, encodes eight FAR-like proteins, and in this review we compare the FAR proteins of C. elegans and parasitic nematodes, and we discuss the suitability of C. elegans as a model system to investigate the biological function(s) of the FAR proteins of parasitic nematodes. PMID- 12592564 TI - Involvement of Toll-like receptor 4 in the embryogenesis of the rodent filaria Litomosoides sigmodontis. AB - To examine the role that lipopolysaccharide (LPS)-like molecules from the filarial intracellular endobacteria Wolbachia might play in the development of filarial infections, a natural infection in the LPS-nonresponsive C3H/HeJ mouse strain was compared to that of the LPS-responsive C3H/HeN mouse strain. C3H/HeN mice have been shown to be susceptible to the rodent filarial nematode Litomosoides sigmodontis, with the development of adult worms including females containing mature microfilariae (first stage larvae) in the uterine tubes. However, free microfilariae are not detected. In this study the worm burden and worm length were not significantly different between the C3H/HeN and C3H/HeJ mice. However, the fertility of worms from CeH/HeJ mice was found to be higher than those from C3H/HeN mice. Significantly, mature microfilariae were found at the site of infection only in C3H/HeJ mice. These results indicate a role for TLR4 signaling in the immune response that inhibits worm embryogenesis and prevents the release of microfilariae or directly kills released microfilariae. PMID- 12592565 TI - Immunopathogenesis of Onchocerca volvulus keratitis (river blindness): a novel role for endosymbiotic Wolbachia bacteria. AB - River blindness is thought to occur as a result of the host response to degenerating microfilariae in the eye. Utilizing a murine model of corneal inflammation (keratitis) to investigate the immune and inflammatory responses associated with river blindness, we recently demonstrated an important role for endotoxin-like products from endosymbiotic bacteria and for activation of Toll like receptor 4 (TLR4). These observations have led to a new understanding of the pathogenesis of this disease PMID- 12592566 TI - Clustered and laminar topographic patterns in rat cerebro-pontine pathways. AB - Novel neuroanatomic approaches for investigating topographic maps at a systems level include combined use of sensitive neural tracing techniques and computerized methods for three-dimensional reconstruction from serial sections. Application of these methods have allowed discovery of new principles of topographic organization in the rat pontine nuclei. The pontine nuclei are intercalated in the large pathways connecting the cerebral cortex to the cerebellum. In rat, cerebropontine projections are characterized by multiple delineated terminal fields. It is generally accepted that these projections are topographically organized. The presence of widespread axonal clusters in the pontine nuclei is typically interpreted to represent a complex scheme of organization. In recent anatomic investigations of somatosensory corticopontine projections in young and adult rats, a somatotopic distribution of axonal clusters, concentrically organized in an inside-out fashion, has been reported. This review summarizes the topographic principles of organization proposed for somatosensory corticopontine projections, and discusses the possibility that widely segregated corticopontine terminal fields are located inside lamellar volumes. This organizational pattern may be explained by mechanisms operative during development, and resembles the patterns of organization previously described in cat and monkey. Possible implications of this architecture are discussed in relation to map transformations from the cerebral cortex to the cerebellum. PMID- 12592567 TI - Different expression of 25-kDa heat-shock protein (Hsp25) in Meckel's cartilage compared with other cartilages in the mouse. AB - The 25-kDa heat-shock protein (Hsp25) is expressed in the cartilage of the growth plate and suggested to function in chondrocyte differentiation and degeneration. Using immunohistochemistry, we examined the temporal and spatial occurrence of Hsp25 in Meckel's cartilage in embryonic mice mandibles, and in other types of cartilage in both embryonic and adult mice. In adults, Hsp25 immunoreactivity was detected in the hypertrophic chondrocytes located in growth plates of long bones and in non-osteogenic laryngeal and tracheal cartilages. No chondrocytes in the resting or proliferating phase exhibited Hsp25 immunoreactivity. In the embryonic mandibles, resting and proliferating chondrocytes in the anterior and intermediate portions of Meckel's cartilage showed Hsp25 immunoreactivity from the 12th day of gestation (E12) through E15, whereas those in the posterior portion showed little or no immunoreactivity. After E16, the overall Hsp25 immunoreactivity in Meckel's cartilage substantially reduced in intensity, and little or no immunoreactivity was detected in the hypertrophic chondrocytes located in the degenerating portions of Meckel's cartilage. The antisense oligonucleotide for Hsp25 mRNA applied to the culture media of the mandibular explants from E10 embryos caused significant inhibition of the development of the anterior and middle portions of Meckel's cartilage. These results suggested that Hsp25 is essential for the development of Meckel's cartilage and plays different roles in Meckel's cartilage from those in the permanent cartilages and the cartilages undergoing endochondral ossification. PMID- 12592569 TI - Homocysteine is embryotoxic but does not cause neural tube defects in mouse embryos. AB - The observation of elevated maternal plasma homocysteine concentration in pregnancies affected by neural tube defects (NTD) suggests that folate metabolism may be disturbed in NTD cases. In addition, studies on the chick embryo suggest that hyperhomocysteinaemia may contribute directly to the development of NTD. In order to test the hypothesis that homocysteine may cause NTD, we cultured mouse embryos in the presence of homocysteine thiolactone during the period of cranial neural tube closure. At doses of 0.5 mM or above, exposure to homocysteine thiolactone caused growth retardation, blisters and abnormalities of somite development. Despite the teratogenic effects of homocysteine we did not detect any increase in the incidence of neural tube defects. Neither was there an effect on the incorporation of thymidine into DNA, a potential marker of alterations in the folate or homocysteine/methionine cycles. These observations suggest that homocysteine is unlikely to be a direct cause of NTD in humans. Rather, the elevated levels of homocysteine in human NTD pregnancies may reflect a disturbance in folate-related metabolism. PMID- 12592568 TI - Localization of intercellular adherens junction protein p120 catenin during podocyte differentiation. AB - To reveal the role of cadherin complex in podocyte differentiation, the present study describes the localization of the cadherin complex, including p120 catenin (p120ctn), in the developing and in the aminonucleoside nephrosis (PAN nephrosis) rat kidney, by immunofluorescence microscopy and immunogold electron microscopy. p120ctn and beta-catenin were co-localized at the apical part of lateral cell membranes in presumptive podocytes of the S-shaped body, and their localization shifted to the basal margin of lateral cell membranes in the capillary loop stage. There was no expression of the cadherin complex at the slit diaphragm, the intercellular junction of mature podocytes. After the regression of the podocyte junctional structure in PAN nephrosis, the cadherin complex was not re-expressed. The dynamic changes in the localization of the cadherin complex suggest that the it plays an important role during podocyte differentiation, including the rearrangement of the intercellular junction and the formation of the slit diaphragm. PMID- 12592571 TI - Intracranial anatomic asymmetry in situs inversus totalis. AB - Situs inversus is a rare condition in which the position of the thoracic and the abdominal viscera are exchanged from the left to the right sides. A possible inversion of normal dominant intracranial anatomy, has however, rarely been discussed. We examine here the gross anatomy of an elderly cadaveric female for a possible "situs inversus" of the intracranial contents. This study has found that many structures commonly dominant on one side in the intracranial compartment were reversed in this specimen. These findings support the concept that a reversal of more commonly found intracranial anatomy may occur in situs inversus totalis, and this should alert the clinician performing invasive procedures in this population. These data will also hopefully provide further insight into possible mechanisms that contribute to situs inversus totalis. PMID- 12592572 TI - Embryos sired by males without accessory sex glands induce failure of uterine support: a study of VEGF, MMP and TGF expression in the golden hamster. AB - To account for reproductive failure induced by surgical deletion of paternal accessory sex glands in the golden hamster in vivo, we studied expression of vegf, FLT-1 (VEGF-R1), FLK-1 (VEGF-R2), MMP and TGF-beta in endometrium of the dam and sired embryos during 5-7 days post coitum by immunohistochemistry, in situ hybridisation, semiquantitative RT-PCR and enzyme-linked immunosorbent assay. Spatiotemporal pattern of vegf expression in the control animals was similar to that reported for intact animals by our group. Removal of paternal ampullary glands did not disturb the normal expression pattern. Removal of ventral prostate glands alone or all accessory sex glands was associated with reduction of vegf transcripts and protein levels in both the embryo and endometrium. FLT-1, FLK-1 and MMP-2 were also reduced. MMP-1 was not changed whereas TGF-beta1 expression was enhanced. There was no expression in endometrium in between implantation sites. Thus the implanted embryos had a trophic effect on growth factor production by the endometrium, and the levels of expression were determined by viability and structural integrity of the conceptus. Based on these findings we concluded that incompetent embryos sired by males without the ventral prostate gland or all accessory sex glands reduced the potential of the uterus to support pregnancy. A negative cycle of events was thus set up and eventually led to premature termination of pregnancies. PMID- 12592570 TI - The lymphoid and immunohaematopoietic tissues of the embryonic brushtail possum ( Trichosurus vulpecula). AB - The lymphoid and immunohaematopoietic tissues of the embryonic and full-term brushtail possums was investigated histologically and immunohistochemically using antibodies to the T- and B-cell markers, CD3, CD5, CD79a and CD79b. No clearly defined thymus, bone marrow, spleen, lymph nodes, gut-associated lymphoid tissues or bronchus-associated lymphoid tissues were observed histologically. The liver was haematopoietic and contained erythrocytic and granulocytic precursors. No mature lymphocytes were observed histologically or detected using antibodies to T and B-cell markers in any of the tissues. These results are consistent with other studies of the early postnatal tissues of other marsupials and support the proposition that neonatal marsupials are substantially reliant on maternal immunological protection at the time of birth and for a significant period of pouch life. PMID- 12592573 TI - Mosaic evolution of neural development in anurans: acceleration of spinal cord development in the direct developing frog Eleutherodactylus coqui. AB - Previous studies have shown that spinal cord development in direct developing frogs of the genus Eleutherodactylus, which have evolutionarily lost the tadpole stage, differs from that in biphasically developing anurans (with the larval and the adult stage separated by metamorphosis). The present study of spinal cord development in Eleutherodactylus coqui provides additional information about neurogenesis, neuronal differentiation and growth analyzed by immunostaining for proliferating cell nuclear antigen (PCNA), in situ hybridization for NeuroD, and morphometric measurements in various developmental stages. Furthermore, spinal cord development in the frogs Discoglossus pictus, Xenopus laevis, and Physalaemus pustulosus, which belong to different anuran families but all exhibit biphasic development, was similarly analyzed. This comparative analysis allows inference of the ancestral anuran pattern of spinal cord development and how it has been modified during the evolution of Eleutherodactylus. All biphasically developing frogs analyzed share a similar pattern of spinal cord development, suggesting that this is ancestral for anurans: after neural tube closure, levels of proliferation and neurogenesis in the spinal cord were low throughout embryogenesis until they were upregulated drastically at early larval stages followed by development of the lateral motor columns. In contrast, no such quiescent embryonic period exists in E. coqui, where rapid growth, high levels of proliferation and neurogenesis, and early formation of lateral motor columns occur shortly after neural tube closure, while other parts of the central nervous system develop more slowly. Thus, spinal cord development has been accelerated during the evolution of Eleutherodactylus relative to the development of other parts of the central nervous system, probably related to the precocious development of limbs in this lineage. PMID- 12592574 TI - Teratogenic effects of suramin on the chick embryo. AB - Suramin, a polysulfonated naphthylamine, has been used for the chemotherapy of trypanosomiasis and onchocerciasis since about the 1920s. Currently, it is also being tested as an anticancer agent. It is hoped that suramin might stop the progression of some kinds of cancer since it has been found to inhibit the proliferation and migration of cells and the formation of new blood vessels. These processes are not only essential for the development and progression of cancer, but also for normal embryonic development. Suramin might, therefore, be a potent teratogen. In the literature, however, we have found only scant information on this subject. In the present study, we demonstrate the teratogenic effects of suramin on chick embryos. Suramin was injected into the coelomic cavity of chick embryos on incubation day (ID) 3. Following reincubation until ID 8, suramin-treated embryos ( n=50) were examined for congenital malformations and compared with a control group ( n=30). The survival rate of suramin-treated embryos was markedly reduced compared with controls (50% vs 90%). Among the 25 survivors the following malformations were recorded: caudal dysgenesia (100%), median facial clefts with hypertelorism (92%), malformations of the aortic arch arteries (88%), hypo-/aplasia of the allantoic vesicle (84%), microphthalmia (52%), abnormalities of the great arterial trunks (44%), unilateral or bilateral cleft lips (40%), heart defects with juxtaposition of the right atrial appendage (36%), persistence of the lens vesicle (32%), median clefts of the lower beak (8%), omphalocele (4%), and cloacal exstrophy (4%). These results show that suramin is a potent teratogen. The possible implications of our findings for human beings and the possible teratogenic mechanisms of suramin are discussed. Use of suramin in experimental teratology might help to clarify the morphogenesis of median facial clefts and of some congenital heart defects. PMID- 12592575 TI - Development of different mast cell types in the opossum Didelphis albiventris. AB - Previous studies have disclosed three types of mast cell in opossums: connective tissue (CTMC), mucosal (MMC), and lymphatic sinus (LSMC). In contrast to most opossum lymph nodes, the mesenteric lymph node is virtually devoid of LSMC, displaying medullary cord CTMC. The present study aimed to describe the development of these mast cell populations. Toluidine blue staining and a histochemical method for demonstrating heparin allowed the identification of immature and mature mast cells. Immature CTMC devoid of detectable heparin were rare until postnatal day 10. Mature CTMC filled with heparin-containing granules became numerous by day 30 to day 40. In the ileum, despite the presence of mature CTMC in the submucosa and mucosa (villus base), immature mast cells first appeared in the villus core by day 65 and adult features were apparent by day 100. In LSMC-containing lymph nodes, immature mast cells were found in lymphatic sinuses by day 10. Clear signs of LSMC differentiation were observed from day 20. Compared with the 10-day value, the mean diameter of cytoplasmic granules at day 40 had doubled and that at day 110 had tripled. In the mesenteric lymph nodes, immature mast cells differentiated into lymphatic sinus CTMC-like cells. After day 80, most of them were located in medullary cords. Weaning and complete maturation of mucosa preceded the differentiation of MMC. In lymph nodes, LSMC differentiation occurred in parallel with the development of the medullary region and deep cortex units. PMID- 12592576 TI - Uptake of antineoplastic agents in pharmacy and hospital personnel. Part I: monitoring of urinary concentrations. AB - OBJECTIVES: There is a risk of adverse health effects for personnel with occupational exposure to antineoplastic agents. The aim of the present longitudinal study was to identify, quantify, and evaluate potential health hazards of occupationally exposed workers in pharmaceutical and oncological departments with central processing units for drug preparation. METHODS: One hundred operatives in 14 German hospital pharmacies and oncological departments underwent biological monitoring by providing urine samples up to four times over a period of 3 years. RESULTS: All antineoplastic agents that were considered (cyclophosphamide, ifosfamide, doxorubicin, epirubicin and platinum from cisplatin and carboplatin), were found in urine samples in up to 40% of participants. CONCLUSIONS: Despite standard safety precautions, such as the use of vertical laminar air flow safety cabinets, and personal protective clothing, incorporation of drugs was detected. Therefore, an environmental monitoring strategy should be developed in order to detect contamination and attempt to improve hygiene during work. PMID- 12592577 TI - Uptake of antineoplastic agents in pharmacy personnel. Part II: study of work related risk factors. AB - OBJECTIVES: This study aimed to find working conditions related to internal exposure of substances handled in centralised cytostatic drug preparation units in hospitals. Recommendations to avoid this uptake should be deduced from the results. METHOD: In a longitudinal study over 3 years, 87 pharmacy technicians and pharmacists of 14 different hospitals in Germany provided 24-h urine samples separately up to three times (three sampling cycles: cycles 1-3) at the end of a working week. Additional samples were taken after 2 days and after at least 3 weeks of absence. Cyclophosphamide and ifosfamide, doxo-, dauno- epi-, and idarubicin, and platinum deriving from cis- and carboplatin were determined in urine samples by gas chromatography/mass spectrometry, liquid chromatography (HPLC) and voltammetry. The following working conditions were assessed by questionnaire: working tasks, different ways that the workbenches were run, cleaning conditions, waste disposal, number of preparations, amount of substances handled, and use of gloves (material, thickness and changing interval). RESULTS: Two-thirds of the subjects showed at least one positive result with regard to all three cycles (56 of initially 87 subjects). Employees who only pass material that is needed for processing are affected, just as are those who only prepare administrations and those alternating in both functions (25% vs. 24.1% vs. 50.6%, respectively). The storage of waste in containers that could be opened to add waste tends to increase the risk of internal exposure of ifosfamide and cyclophosphamide (odds ratios (95% confidence interval): 0.08 (0.013-0.5) and 0.19 (0.03-1.12), respectively). The amount handled and number of preparations of cyclophosphamide for "manufacturers" were associated with internal exposure of cyclophosphamide (28.04 (1.75-448.74) and 1.22 (1.03-1.44), respectively). The total number of preparations handled by assistants seemed to increase the risk of intake of any of the substances under study [1.04 (1.00-1.08)]. CONCLUSION: Since employees who pass materials are affected in the same way as those who prepare administrations, both have to be included in reviewing protective measures. Further studies must be carried out to verify the generated hypotheses of factors related to internal exposure found in this study. PMID- 12592578 TI - Methyl isobutyl ketone and methyl ethyl ketone in urine as biological markers of occupational exposure to these solvents at low levels. AB - OBJECTIVE: To examine whether unmetabolized methyl isobutyl ketone in urine is a useful marker of low-level occupational exposure to this ketone solvent, as is the case for methyl ethyl ketone. METHODS: The study was conducted in the second half of a working week. In total, 27 furniture-making workers (19 men and eight women) and 11 non-exposed controls (six men and five women) volunteered to participate in the study. Time-weighted average (TWA; 8-h) concentration of vapors of several solvents in air, including methyl isobutyl ketone (MIBK-A) and methyl ethyl ketone (MEK-A) was monitored by diffusive sampling. Urine samples collected at the end of the shift were subjected to head-space gas chromatography (GC) analysis for the unmetabolized solvents (i.e., MIBK-U and MEK-U). The relationship between the concentration of the solvent vapor and the corresponding solvent level in urine was examined by simple as well as multiple regression analysis. RESULTS: The exposures to MIBK and MEK were well below the current occupational exposure limit levels, but the maximum levels of exposure to toluene and ethylbenzene were around the corresponding exposure limit. The correlation of the TWA concentration of the solvent in air with the concentration of the corresponding solvent in the end-of-shift urine sample was significant both for MIBK and for MEK, and the correlation coefficient was larger for MIBK than for MEK. The slope in the exposure-excretion regression line was almost twice as steep for MEK than for MIBK, possibly due to the difference in water solubility. Approximately 0.12% of MIBK absorbed in the lungs will be excreted into urine, whereas the yield for MEK was somewhat higher (0.19%). CONCLUSION: MIBK in urine is a good marker of exposure to MIBK, as in the case of MEK in urine for MEK exposure. PMID- 12592579 TI - Increased incidence of haematological cancer among physicians in a University Hospital. AB - OBJECTIVES: The oral report of eight cases of cancer over a period of 3 years among physicians working in a French University Hospital led us to conduct a retrospective cohort study to compare the incidence of cancer in these physicians with that of the general population living in the same area. METHOD: The cohort consisted of 940 physicians (72% male, 28% female) who had worked for at least 1 year in this hospital between 1945 and 1994 (a total of 10,693 person-years). The incidence of cancer among physicians was compared with that of the general population using a local cancer registry which has recorded all cancer cases occurring in the Department between 1979 and 1994. Standardised incidence ratio (SIR) was calculated for all causes of cancer and site by site. RESULTS: Although the global incidence of cancer did not differ from that of the general population (SIR=0.97; 95% confidence interval (CI) 0.59-1.5), a significantly increased incidence of haematological malignancy was found among physicians (SIR=5.45; 95% CI 2-11.9). CONCLUSIONS: These findings pointed out the risk of lymphatic and haematopoietic cancer among physicians who started working after 1945 when the first rules pertaining to protection from ionising radiation came into effect. However, limitations of this study such as limited statistical power and lack of individual exposure data should be considered in the interpretation of these findings. Possible aetiological factors responsible for these risks are discussed. Education about general safety protection programmes against carcinogenic risk factors including physical, biological and chemical agents still remains a priority among physicians. PMID- 12592580 TI - Symptoms and sense of coherence--a follow-up study of personnel from workplace buildings with indoor air problems. AB - OBJECTIVES: The aim was to study prevalence and change of symptoms in buildings with suspected indoor air problems in relation to sense of coherence (SOC), a psychological measure of a life attitude. METHODS: A cohort of 194 subjects initially working in 19 Swedish buildings with indoor environmental problems was followed from 1988 to 1998. Information on 16 symptoms compatible with sick building syndrome (SBS) was gathered by an initial questionnaire mailed between 1988 and 1992. The same symptom questionnaire, as well as Antonovsky's SOC, was administered in a postal follow-up study in 1998. The prevalence of symptoms and the change (incidence) plus reminiscence of symptoms were calculated for individual symptoms and a total symptom score (SC). Bivariate analyses, as well as multiple linear and logistic regression analyses, were applied and adjusted for age, gender, history of atopy and tobacco smoking. RESULTS: SBS was more common in women, younger subjects and those with a history of atopy. A low SOC was related to a higher prevalence of ocular, nasal, and throat symptoms, tiredness, and headache. In addition, subjects with a low SOC developed more symptoms during the follow-up period. Women had a lower SOC value, but there was no relation between SOC and age, smoking, doctor's diagnosed asthma or a history of atopy. Subjects leaving the problem buildings during the follow-up period had a decrease in symptoms and were more often non-smokers, but had the same mean SOC score as those remaining in the same workplace. CONCLUSIONS: The study indicates that SOC can detect personal vulnerability in relation to suspected environmental stress. Symptoms reported in the buildings with suspected indoor air problems are partly reversible, as indicated by the reduction of symptoms among those leaving these buildings. A multi-disciplinary approach including personality aspects, allergic disorders and indoor exposures should be applied in investigations of buildings with suspected indoor air problems. PMID- 12592581 TI - The effects of neck-shoulder pain development on sensory-motor interactions among female workers in the poultry and fish industries. A prospective study. AB - OBJECTIVES: The purpose of this prospective laboratory study was to follow newly employed workers in the fish or poultry industry for 6 months and investigate possible changes in sensory manifestations and motor performance during low load, repetitive work simulation. It was investigated whether the changes were an effect of employment duration and of development of neck-shoulder complaints. METHODS: Twelve newly employed female filleting employees without any sign of neck-shoulder tenderness/pain at the time of employment of 0 months took part in two laboratory recording sessions planned within 1 month of employment and after 6 months of employment at the plant. After 6 months, six workers out of 12 had developed pain and/or tenderness in the neck-shoulder region. The recording sessions evaluated sensory-motor aspects by measuring pressure pain threshold, work-task timing, cutting forces, surface electromyographic activity of four shoulder muscles, displacement of the centre of pressure, and 3D movements of the arm and trunk during simulation of low load, repetitive filleting. RESULTS: Effects due to the duration of employment were observed in both groups, i.e. decreased sensibility to pressure, decrease in the duration of the work cycle, increased arm starting position with respect to the upright position, and decreased range of motion of the arm and trunk (P<0.05) after 6 months. Among the workers with neck-shoulder complaints, increased sensibility to pressure, lower force level, higher electromyographic activity, decreased amplitude of arm movement, and increased trunk posture and movement amplitude (P<0.05) were observed, compared with workers without complaints. CONCLUSIONS: Differences in terms of sensory manifestations and motor control strategy were seen after 6 months of employment with or without neck-shoulder complaints. In general, changes in sensory manifestations and motor control strategy after 6 months work were most likely of importance, as they underlined a learning process as employment duration increased. Moreover, the present sensory-motor changes observed among workers with neck-shoulder complaints highlighted the potential physical risk factors associated with low load, repetitive work. PMID- 12592582 TI - Web-based training in occupational medicine. AB - OBJECTIVES: The aim of our project was to develop and evaluate an interactive computer-based approach to teach medical students in occupational medicine. To enhance interest in occupational medicine the major focus was on clinical and practical aspects of occupational medicine. METHODS: The computer program was designed in HTML and JavaScript. It presents a guided tour of the patient's case history followed by information on practice and theory of occupational medicine. The program was integrated into the curriculum during the summer term of 1999 and the following winter term. To assess the effectiveness and acceptability of the program we asked students to rate the program on an 18-item questionnaire. RESULTS: Overall, 287 students participated in the evaluation of the program. The participants highly recommended the program structure and had no difficulties in handling the program. This was independent of the computer experience of the students. The evaluation showed that it was possible to enhance interest in occupational medicine by using "virtual patients". CONCLUSION: The program represents a student-orientated learning tool that points out clinical and practical aspects of occupational medicine. The availability via the Internet allows the application as a self-learning tool as well as a teaching tool for the medical curriculum. PMID- 12592583 TI - Urine mutagenicity of farmers occupationally exposed during a 1-day use of chlorothalonil and insecticides. AB - OBJECTIVES: A study was conducted to evaluate the genotoxic impregnation consecutive to a 1-day open-field spraying of pesticides. METHODS: From 14 farmers (five smokers and nine non-smokers), three urine samples were collected at the end of the spraying season: the morning (S1) of the day of spraying, the evening (S2) and the morning (S3) of the following day. A fourth sample (S0) was obtained before the pesticide-handling period. Mutagenicity of urine extracts was evaluated with the Ames test, using strains TA97a, TA98, TA100 and TA102, with and without S9 mix. RESULTS: The ratio of induced vs spontaneous revertants (induction ratio) was > or =2 in five farmers (including three smokers), with only one strain responding in each. Applying the SALM software proposed by Kim and Margolin in combination with the ANOVA-Dunnett test on crude data (number of revertants), urine extracts were found to be mutagenic on at least one Salmonella strain in 57% and 96% of non-smokers and smokers, respectively. The proportion of mutagenic responses tended to increase from S1 to S3 (not statistically significant) in non-smokers only. Finally, there were no relationships between the relative changes in the number of revertants (adjusted for urine concentration) and any exposure parameters available: area sprayed, number of tanks prepared and time free of exposure to any pesticide. CONCLUSIONS: The lack of significant relationships between urine mutagenicity and exposure data argues against a direct role of the pesticides sprayed, on this impregnation. This result should be considered with caution since the number of farmers involved may limit the significance of the study. PMID- 12592584 TI - Exposure to diesel exhaust enhances total IgE in non-atopic dockers. AB - OBJECTIVES: Diesel exhaust particles (DEPs) containing polycyclic aromatic hydrocarbons stimulate the formation of IgE in humans following single and acute exposure. The aim of the present study was to ascertain whether long-standing occupational exposure to DEPs carries a risk of enhanced serum IgE, and of rhinitis or asthma. METHODS: In this cross-sectional study, findings in 76 dockers were compared with those in 63 reference subjects. Among the dockers, drivers and laborers were exposed to diesel emission from forklifts or trucks in the ship-holds, where benzopyrene levels averaged 4.9 ng/m(3). Serum IgE levels were measured by the UNICAP method. Atopy, an evident source of high IgE levels, was assessed by the Phadiatop test. The subjects' clinical and occupational histories were collected. Interval variables were analyzed with Student's t- and Levene's F-tests. The odds ratio (OR) with the 95% confidence interval (CI) was obtained by the exact method at univariate and multivariate (logistic regression) analysis. RESULTS: In view of the large difference in serum IgE (P=0.00001) and the prevalence of respiratory diseases ( P=0.009) between atopic and non-atopic subjects, we analyzed their data separately. For non-atopic subjects, the risk of presenting high IgE was significantly higher (OR=11.4; CI=1.44-526; P=0.013) and the risk of respiratory disease was significantly lower (OR=0.09; CI=0.00-0.73; P=0.016) in drivers and laborers as a whole than in the reference subjects. None of the ORs was significant among atopic individuals. CONCLUSIONS: In non-atopic dockers, long-standing exposure to DEPs at concentrations similar to those in heavily polluted cities increased serum IgE levels but not the incidence of rhinitis or asthma. PMID- 12592585 TI - Health and psychosocial work characteristics of employees who did or did not consult the general practitioner in relation to work. AB - OBJECTIVES: To examine how health problems and psychosocial work characteristics are associated with having visited or not having visited the general practitioner (GP) in relation to work. METHODS: Baseline self-reported data of the Maastricht Cohort Study about fatigue at work were used to gather information about fatigue (Checklist Individual Strength), presence of at least one long-term disease, likelihood of having a mental illness (General Health Questionnaire), and psychosocial work characteristics (psychological job demands, decision latitude and social support at work, as measured with the Job Content Questionnaire). The cohort participants indicated having visited (VISITORS) or not having visited (NON-VISITORS) the GP in relation to work. Differences between these groups were measured by multiple logistic regression analyses. RESULTS: When compared with VISITORS and taking into account the influence of sociodemographic characteristics and mutual associations between reported health problems and psychosocial work characteristics, a lower percentage of NON-VISITORS reported at least one long-term disease and NON-VISITORS presented lower levels of fatigue and psychological job demands, and higher levels of decision latitude and social support at work. CONCLUSIONS: The results of this study indicate that NON VISITORS reported fewer health problems and a more positive perception of their work environment than VISITORS did. Interrelationships between psychosocial work characteristics and health variables should be taken into consideration when studying their associations with visiting the GP in relation to work. PMID- 12592586 TI - Persistent chlorinated pesticides and intra-uterine foetal growth retardation: a possible association. AB - OBJECTIVE: To examine the association between DDT (dichlorodiphenyl trichloroethane) and HCH (hexachlorocyclohexane) exposure and intra-uterine growth retardation (IUGR, <10th percentile of birth weight for gestational age). METHOD: We detected p,p'-DDT, o,p'-DDT, p,p'-DDD, p,p'-DDE and alpha-HCH, beta HCH, gamma-HCH, delta-HCH in maternal blood, placenta and cord blood, collected at parturition, from mothers with IUGR babies ( n=30) and from those with babies of normal weight ( n=24), using gas-liquid chromatography equipped with electron capture detector ((63)Ni). The adjusted odds ratios (ORs) for these pesticides in mothers and infants were determined by multiple logistic regression. RESULTS: There were statistically significant associations ( P<0.05) between maternal blood levels of alpha-HCH (OR=1.22; 95% CI: 1.02-1.46), gamma-HCH (OR=1.38; 95%CI: 1.05-1.80), delta-HCH (OR=1.61; 95% CI: 1.01-2.54), total HCH (OR=1.07; 95% CI: 1.01-1.13) and p,p'-DDE (OR=1.21; 95%CI:1.03-1.42) and IUGR after adjustment for potential confounders. Also, significant association (P<0.05) between cord blood levels of gamma-HCH (OR=1.14; 95% CI: 1.00-1.31), delta-HCH (OR=1.31; 95% CI: 1.00-1.75), total HCH (OR=1.07; 95% CI: 1.00-1.14) and IUGR were found after adjustment for potential confounders. A significant negative correlation between body weight of newborn babies and p,p'-DDE in maternal blood (r= -0.25; P<0.05) and delta-HCH and p,p'-DDE in the cord blood (r= -0.27 and 0.26; P<0.05) was noticed after gestational age had been accounted for. CONCLUSION: Exposure of pregnant women to organochlorine pesticides may increase the risk of IUGR, which is a contributing factor for infant mortality in India. PMID- 12592587 TI - Cardiovascular function of workers exposed to carbon disulphide. AB - OBJECTIVES: The aim of the study was to verify that exposure to carbon disulphide (CS(2)) up to 10 ppm results in a negative inotropic effect on cardiovascular function. METHODS: In a cross-sectional study a total of 325 workers exposed to CS(2) in the rayon-producing industry and 179 controls from the same plants were examined. The exposure assessment was based on personal air sampling and biological monitoring (2-thiothiazolidine-4-carboxylic acid (TTCA) in urine) for all persons. The examination consisted of a standardised questionnaire, physical examination, assessment of body fat mass, ergometric test with the determination of work capacity at heart rates of 100, 130, 150 and 170 beats/min, and electrocardiography. RESULTS: The mean external exposure was 6.04 ppm CS(2) (range: 0.03-91.08); the mean internal exposure was 1.14 mg TTCA/g creatinine (range: 0.02-11.50). The workers exposed to CS(2) showed better physical fitness. The diameters of the left heart chamber of the exposed persons were not significantly different when compared with occupationally non-exposed workers, but there was a tendency of increasing diameters for the exposed employees. In the multiple linear regression the diameters showed physiologically plausible correlations with the body mass index, body fat mass, alcohol consumption, and physical fitness, but not, however, with the exposure, neither with the exposure group in all persons nor with the internal or external exposure within the exposed workers. CONCLUSIONS: In this study, differences in the heart chamber diameters between exposed persons and controls could not be confirmed. Differences in physical fitness and constitution can explain differences in heart chamber diameters. PMID- 12592589 TI - Intracartilaginous haemorrhagic lesions in strangulation? AB - This case control study was designed to investigate if laryngeal haemorrhages occur in cases of strangulation and whether these lesions are specific to strangulation. In the study 30 larynges from victims of fatal strangulation were examined (7 cases of manual strangulation, 12 cases of ligature strangulation, 11 cases of combined manual and ligature strangulation). The control group comprised 40 cases of death without any neck injuries and another group consisted of 5 cases of death caused by trauma with findings of non-strangulation neck injuries. In all the groups, only four solitary haemorrhages (two cases, one control, one non-strangulation neck injury) were observed that did not occur in the proximity of areas of blood accumulation. The results of our investigation suggest that histological evidence of blood accumulation or of haemorrhages in thyroid cartilage is not a reliable criterion to distinguish between haemorrhagic lesions due to strangulation and other types of blood accumulation or artefacts. PMID- 12592588 TI - Immunohistochemical demonstration of the amphetamine derivatives 3,4 methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) in human post-mortem brain tissues and the pituitary gland. AB - Abuse of amphetamine derivatives such as 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) is an important issue in current forensic practice and fatalities are not infrequent. Therefore, we investigated an immunohistochemical method to detect the amphetamine analogues MDMA and MDA in human tissues. For the staining procedure, the Catalysed Signal Amplification (CSA) method using peroxidase (HRP) provided by Dako and specific monoclonal antibodies were used. Appropriate controls for validation of the technique were included. The distribution of these designer drugs was studied in various brain regions including the four lobes, the basal ganglia, hypothalamus, hippocampus, corpus callosum, medulla oblongata, pons, cerebellar vermis and, additionally, in the pituitary gland. A distinct positive reaction was observed in all cortical brain regions and the neurons of the basal ganglia, the hypothalamus, the hippocampus and the cerebellar vermis but in the brainstem, relatively weak staining of neurons was seen. The reaction presented as a mainly diffuse cytoplasmic staining of the perikaryon of the neurons, and often axons and dendrites were also visualised. In addition, the immunoreactivity was present in the white matter. In the pituitary gland, however, distinct immunopositive cells were observed, with a prominent heterogeneity. The immunohistochemical findings were supported by the toxicological data. This immunostaining technique can be used as evidence of intake or even poisoning with MDMA and/or MDA and can be an interesting tool in forensic practice when the usual samples for toxicological analysis are not available. Furthermore, this method can be used to investigate the distribution of these substances in the human body. PMID- 12592590 TI - Immunohistochemical localization of mast cells as a tool for the discrimination of vital and postmortem lesions. AB - In wounds that are inflicted at least 60 min before death, histamine levels can increase up to 100%. This functional effect might have a morphometric counterpart. Mast cells play a crucial role in acute inflammatory reactions and in the healing process of wounds. Therefore, the density of these cells was immunohistochemically assessed in tissue from 20 healthy controls (Group 1), 20 vital skin lesions (Group 2) (age range: a few seconds to 1 h), and 20 postmortem lesions (Group 3). A piece of skin close to the vital lesion was also obtained from the homolateral part of the body (Group 4). Mast cell density was significantly higher at the level of the vital lesions (11.28+/-2.44) than elsewhere (healthy controls 7.66+/-1.27, postmortem lesions 4.13+/-1.46, skin close to the vital lesions 4.88+/-1.59). No differences were found between the values assessed in the skin samples close to the vital lesions and in those in the postmortem lesions. Therefore, mast cell richness in the vital lesions exhibited a proportional morphological correlation with previously detected histamine values in cutaneous vital lesions. These results suggest that the detection of mast cells with immunohistochemical techniques can lead to a high level of discrimination (based on statistical data) between antemortem and postmortem lesions. This method could also be used to ascertain the vitality of lesions. PMID- 12592591 TI - Morphology of experimental assault rifle skin wounds. AB - Finnish forensic experts who had performed investigations of victims of alleged political violence in Kosovo, in the Federal Republic of Yugoslavia, under the mandate of the European Union, carried out experimental shooting in Finland to confirm observations made during the earlier forensic investigation. Experimental shooting can be of benefit for autopsy conclusions, because the wounding potential of military weapons differs from that of non-military weapons. Assault rifle gunshot wounds were inflicted upon anaesthetised swine from various distances and angles and with variable shielding of the skin. The morphology of the skin wounds was studied and post-mortem changes were documented while the wounds were being observed in cool and room temperature conditions for 13 days. Large variation was found in the size, form, and regularity, and in the presence and width of the abrasion zone of entrance and exit wounds, in addition to secondary wounds. The maximum diameter of entrance wounds varied between 4 and 40 mm and that of exit wounds between 10 and 110 mm. The width of the abrasion zone surrounding entrance wounds ranged from 2 to 11 mm. Extreme care and caution are needed when drawing conclusions with regard to cases of multiple gunshot injuries, and especially when post-mortem changes are considerable. PMID- 12592592 TI - Extending STR markers in Y chromosome haplotypes. AB - Two multiplex reactions were developed to amplify 16 Y-STRs (DYS19, DYS385, DYS389 I and II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, GATA A7.1, GATA A7.2, GATA A10, GATA C4, GATA H4). Here we extend previous population studies done in a sample from northern Portugal for the GATA A7.1, GATA A7.2, GATA C4 and GATA H4 loci. A total of 199 different haplotypes identified by the 16 Y-STR markers were observed in a sample of 208 male individuals, of which 190 were unique and 9 were found twice. The overall haplotype diversity was 0.9996. The haplotype diversity of the Y-STR set composed of the 8 new markers is higher than the Y-STR core set included in the Y-STR haplotype reference database. Sequence structure of new alleles for GATA C4 and GATA H4 is reported. The usefulness of the inclusion of this new set of Y-STRs in forensic casework was also assessed. The increase in haplotype diversity with the addition of any new Y STR marker to the 8 Y-STR core set is dependent not only on the gene diversity (positively) but also (negatively) on the degree of gametic association between the markers and the haplotypes previously defined. For instance, in our sample the addition of the DYS437, DYS438 and GATA A7.2 to a 13-locus set increased haplotype diversity only by 0.0001. PMID- 12592593 TI - Haplotype analysis with 14 Y-STR loci using 2 multiplex amplification and typing systems in 2 regional populations in Japan. AB - In this study 14 Y-STR loci (DYS393, DYS19, DYS391, DYS437, DYS435, DYS439, DYS389II, DYS438, DYS436, DYS390, Y-GATA-H4, DYS385, Y-GATA-A7.1 and DYS392) were analysed in 207 Japanese males from Honshu (main island of Japan, Nagoya City) and 87 Japanese males from Okinawa (southernmost islands of Japan) using two multiplex PCR typing systems, a novel 10-plex amplification system and a new commercially available 6-plex typing kit which had two loci in common. The allele frequency distributions were similar at almost all of the 14 loci. Of the haplotypes observed, 244 were unique in both Japanese populations and 17 haplotypes were observed more than once but the 2 populations shared only 7 haplotypes. The haplotype diversities for the 14 loci were 0.9987 and 0.9976 in Honshu and Okinawa Japanese, respectively. The haplotype analysis at 14 Y-STR loci would be useful for personal identification in forensic fields and for population genetics because of the high divergence of these haplotypes. PMID- 12592594 TI - Interpreting DNA mixtures with the presence of relatives. AB - The assessment of DNA mixtures with the presence of relatives is discussed in this paper. The kinship coefficients are incorporated into the evaluation of the likelihood ratio and we first derive a unified expression of joint genotypic probabilities. A general formula and seven types of detailed expressions for calculating likelihood ratios are then developed for the case that a relative of the tested suspect is an unknown contributor to the mixed stain. These results can also be applied to the case of a non-tested suspect with one tested relative. Moreover, the formula for calculating the likelihood ratio when there are two related unknown contributors is given. Data for a real situation are given for illustration, and the effect of kinship on the likelihood ratio is shown therein. Some interesting findings are obtained. PMID- 12592596 TI - Population genetic comparisons of three X-chromosomal STRs. AB - The X-chromosomal short tandem repeats (STRs) DXS6800, DXS101 and DXS8377 were analysed in male and female population samples from Germany and Austria using a PCR multiplex approach. We investigated 135 family trios from Innsbruck (Austria) and surrounding areas and 50 families and further male and female samples from Ulm (Germany) and surrounding areas. The comparisons of the allele frequencies gave similar distributions for Innsbruck and Ulm although minor variations were found for some alleles. Additionally, some differences were found when comparing the allele frequencies of the male and female samples independently. The forensic efficiency values demonstrate that especially DXS101 and DXS8377 are highly informative markers for kinship analysis and deficiency cases. Based on the investigated meiotic events no new mutations were detected. PMID- 12592599 TI - [Guideline-assisted suggestion for training courses in gastrointestinal endoscopy]. PMID- 12592595 TI - Studies on mRNA expression of basic fibroblast growth factor in wound healing for wound age determination. AB - We investigated the mRNA expression of basic fibroblast growth factor (bFGF) for wound age determination during dermal, cerebral, hepatic and renal wound healing in mice. The bFGF mRNA expression in the injured skin peaked at 1 h and was detected in epidermal cells, fibroblasts, endothelial cells and neutrophils. In the injured cerebrum the expression increased from 1 h and peaked at 48 h. In the intact cerebrum, bFGF was detected exclusively in the endothelial cells, whereas it was also detected in astrocytes during wound healing. Time-dependent expression of bFGF mRNA in skin and cerebrum was considered to be useful for wound age determination. On the other hand it was suggested that bFGF mRNA in astrocytes could be a vital sign of the acute phase. In hepatic and renal injuries, however, bFGF mRNA expression increased slightly in endothelial cells at 24 h, in neutrophils of the liver and in the glomeruli of the kidneys. PMID- 12592597 TI - Procalcitonin is a valid marker of infection in decompensated cirrhosis. AB - BACKGROUND/AIMS: Bacterial infections are life-threatening complications in cirrhosis and early diagnosis is mandatory. Procalcitonin, a 116 amino acid propeptide of calcitonin, is an early marker of infection. The aim was to evaluate prospectively procalcitonin in the diagnosis of bacterial infection in cirrhosis. 127 patients with liver cirrhosis were analysed and stratified into three groups according bacteriological and morphological findings; decompensated patients with (group I = 36) and without (group II = 64) infection, and 27 non decompensated and non-infected (group III). METHODS: Diagnosis of infection was made using standard criteria. Serum procalcitonin, tumour necrosis factor alpha, interleukin-6 and C-reactive protein were measured using commercially available methods. RESULTS: PCT serum levels were significantly different between group I (2.8 ng/ml [0.4 - 20.4]), group II (0.6 ng/ml [0.1 - 5.9]) and group III (0.4 ng/ml [0.1 - 1.2]), respectively. Levels above 0.58 ng/ml had a sensitivity of 92 % and specificity of 78 % for the diagnosis of infection and were associated with a 50 % mortality in the first two months. Interleukin-6, tumour necrosis factor alpha and C-reactive protein were less sensitive and specific for the diagnosis of infection. CONCLUSION: In decompensated cirrhosis procalcitonin serum levels provided the most sensitive and specific tool for the initial diagnosis of bacterial infection. PMID- 12592598 TI - [Diagnosis of a high-grade B-cell lymphoma of the small bowel by means of wireless capsule endoscopy]. AB - Wireless capsule endoscopy is a new diagnostic procedure to detect diseases of the small bowel, in particular in patients with obscure gastrointestinal bleeding. We report on a 76-year-old patient who presented at our hospital with signs of gastrointestinal bleeding and hypochromic anaemia. Bidirectional endoscopy, push enteroscopy, angiography as well as small bowel barium enema rendered no pathologic findings. Wireless capsule endoscopy demonstrated a polypous space in the cantral area of the small bowel. Subsequent intraoperative endoscopy revealed a subtotal stenosing tumour of the upper jejunum which was resected. Based on the histology a high-grade B-Cell lymphoma was diagnosed. The postoperative staging examinations did not indicate any further extranodal involvement. PMID- 12592600 TI - [Ogilvie's syndrome: a rare cause of the acute abdomen]. AB - Ogilvie's syndrome (acute colonic pseudo-obstruction) is a rare clinical disease characterized by segmental distension of the proximal colon caused by a paralysis without mechanic obstruction. It may be a sequel of underlying neurological, medical or surgical disease. Risk factors are respiratory decompensation, electrolyte disturbances and different drugs. A special kind is the primary idiopathic pseudoobstruction with a high risk of perforation or necrosis. Especially elderly patients (> 70 years) with cardiovascular or neurologic diseases and accordant drugs are concerned. Clinical symptoms are progressive abdominal distension and abdominal pain like an acute abdomen. The differential diagnosis of a mechanic ileus is important for further treatment. This case report should draw attention to this rare disease.